PMID- 19280263 TI - Feasibility and efficacy of accelerated weekly concomitant boost postoperative radiation therapy combined with concomitant chemotherapy in patients with locally advanced head and neck cancer. AB - BACKGROUND: The aim of this study was to assess feasibility and efficacy of weekly concomitant boost accelerated postoperative radiation therapy (PORT) with concomitant chemotherapy (CT) in patients with locally advanced head and neck cancer (LAHNC). METHODS AND MATERIALS: Conformal or intensity-modulated 66-Gy RT was performed in 5.5 weeks in 40 patients. Cisplatin was given at days 1, 22, and 43. Median follow-up was 36 months. RESULTS AND DISCUSSION: Grade 3 mucositis, dysphagia, and erythema was observed in ten (25%), nine (23%), and six (13%) patients, respectively. Grade 3 or more anemia was observed in two (6%) patients, and leukopenia in five (13%) patients. No grade 3 or 4 thrombocytopenia was observed. Grade 3 nephrotoxicity was observed in one patient (3%). No treatment related mortality was observed. Grade 2 or more xerostomia and edema were observed in ten (25%) and one (3%) patient, respectively. Locoregional relapse occurred in eight patients, and seven patients developed distant metastases. Median time to locoregional relapse was 6 months. Three-year overall, disease free survival, and locoregional control rates were 63%, 62%, and 81%, respectively. Multivariate analysis revealed that the only prognostic factor was nodal status. CONCLUSION: Reducing overall treatment time using accelerated PORT/CT by weekly concomitant boost (six fractions per week) combined with concomitant cisplatin CT is easily feasible with acceptable morbidity. PMID- 19280264 TI - Neoadjuvant chemotherapy in invasive lobular carcinoma may not improve rates of breast conservation. AB - BACKGROUND: Patients with invasive lobular carcinoma (ILC) experience a lower pathological complete response rate to neoadjuvant chemotherapy than patients with invasive ductal carcinoma. This study was intended to evaluate the impact of neoadjuvant chemotherapy in ILC on breast-conserving surgery (BCS) rates. METHODS: Two-hundred eighty-four consecutive patients with pure ILC treated between May 1998 and September 2006 were reviewed. Surgical procedures and long term outcomes were compared between patients receiving neoadjuvant chemotherapy and those receiving surgery first. RESULTS: Neoadjuvant chemotherapy was administered to 84 patients; 200 patients underwent surgery first. The mean tumor size in the neoadjuvant group (4.9 cm) was significantly larger than in patients who underwent surgery first (2.5 cm, p < 0.0001). In the neoadjuvant group, clinical complete response was seen in 10% and partial response in 59%. Overall BCS rates were 17% in the neoadjuvant group compared with 43% in the surgery first group (p < 0.0001). When controlled for initial tumor size, there was no difference (all p > 0.05) between the groups in terms of (1) the proportion of patients who underwent an initial attempt at BCS, (2) rate of failure of BCS or (3) the proportion of patients undergoing BCS as their final procedure. With a mean follow-up of 47 months, local recurrence (LR) rates were similar between the two groups (1.2% versus 0.5%, p = 0.5). CONCLUSION: The use of neoadjuvant chemotherapy does not increase the rates of breast conservation in patients with pure ILC. PMID- 19280265 TI - Neutral models with generalised speciation. AB - Hubbell's neutral theory claims that ecological patterns such as species abundance distributions can be explained by a stochastic model based on simple assumptions. One of these assumptions, the point mutation assumption, states that every individual has the same probability to speciate. Etienne et al. have argued that other assumptions on the speciation process could be more realistic, for example, that every species has the same probability to speciate (Etienne, et al. in Oikos 116:241-258, 2007). They introduced a number of neutral community models with a different speciation process, and conjectured formulas for their stationary species abundance distribution. Here we study a generalised neutral community model, encompassing these modified models, and derive its stationary distribution, thus proving the conjectured formulas. PMID- 19280266 TI - Modeling proximal tubule cell homeostasis: tracking changes in luminal flow. AB - During normal kidney function, there are routinely wide swings in proximal tubule fluid flow and proportional changes in Na(+) reabsorption across tubule epithelial cells. This "glomerulotubular balance" occurs in the absence of any substantial change in cell volume, and is thus a challenge to coordinate luminal membrane solute entry with peritubular membrane solute exit. In this work, linear optimal control theory is applied to generate a configuration of regulated transporters that could achieve this result. A previously developed model of rat proximal tubule epithelium is linearized about a physiologic reference condition; the approximate linear system is recast as a dynamical system; and a Riccati equation is solved to yield the optimal linear feedback that stabilizes Na(+) flux, cell volume, and cell pH. The first observation is that optimal feedback control is largely consigned to three physiologic variables, cell volume, cell electrical potential, and lateral intercellular hydrostatic pressure. Parameter modulation by cell volume stabilizes cell volume; parameter modulation by electrical potential or interspace pressure act to stabilize Na(+) flux and cell pH. This feedback control is utilized in a tracking problem, in which reabsorptive Na(+) flux varies over a factor of two, in order to represent a substantial excursion of glomerulotubular balance. The resulting control parameters consist of two terms, an autonomous term and a feedback term, and both terms include transporters on both luminal and peritubular cell membranes. Overall, the increase in Na(+) flux is achieved with upregulation of luminal Na(+)/H(+) exchange and Na(+)-glucose cotransport, with increased peritubular Na(+)-3HCO(3)(-) and K(+)-Cl(-) cotransport, and with increased Na(+), K(+) ATPase activity. The configuration of activated transporters emerges as a testable hypothesis of the molecular basis for glomerulotubular balance. It is suggested that the autonomous control component at each cell membrane could represent the cytoskeletal effects of luminal flow. PMID- 19280267 TI - Mid-term follow-up after sleeve gastrectomy as a final approach for morbid obesity. AB - BACKGROUND: In previous publications, we demonstrated the safety and short-term efficacy of laparoscopic sleeve gastrectomy (LSG) as a final step in the treatment of morbid obesity (MO). This study aimed to assess the mid-term efficacy of LSG. METHODS: We performed a retrospective review of a prospectively collected database. Between November 2004 and January 2007, 130 consecutive patients underwent LSG as a final procedure to MO. Data including patient demographics, operative time, length of hospital stay, complications, preoperative body mass index (BMI), complications, and weight loss at 3, 6, 12, 18, and 24 months were recorded and analyzed. RESULTS: The mean age was 45.6 (range: 12-79) years while the mean BMI was 43.2 (range: 30.2-75.4) kg/m(2). The mean operative time was 97 (range, 58-180) min and all operations were completed laparoscopically. The mean hospital stay was 3.2 (range, 1-19) days with zero mortality in this series. One patient (0.7%) had leakage at the stapler line, while four patients (2.8%) developed trocar site infection. Three patients (2.1%) complained of symptoms of gastroesophageal reflux disease (GERD), three patients (2.1 %) developed symptomatic gallstones, and trocar site hernia was present in one (0.7%) patient. The mean weight loss was 21, 31.2, 37.4, 39.5, and 41.7 kg at 3, 6, 12, 18, and 24 months, respectively, while the mean BMI decreased to 36.9, 32.8, 29.5, 28, and 27.1 at 3, 6, 12 18, and 24 months, respectively. Percent of excess weight loss (%EWL) was 33.1, 50.8, 62.2, 64.4, and 67.9 at 3, 6, 12, 18, and 24 months, respectively. CONCLUSIONS: LSG is a safe and effective surgical procedure for the morbidly obese up to 2 years. Excess body weight loss seems to be acceptable at 2 years postoperatively. PMID- 19280268 TI - Expression of cytokine signaling genes in morbidly obese patients with non alcoholic steatohepatitis and hepatic fibrosis. AB - BACKGROUND: White adipose tissue (WAT) from visceral adiposity plays an important role in the pathogenesis of non-alcoholic steatohepatitis (NASH). Development of NASH and its progression to fibrosis is partially due to cytokines and adipokines produced by WAT. The aim of this study was to assess the association of hepatic fibrosis and NASH by evaluating the intrinsic differences in the inflammatory cytokine signaling in the visceral adipose tissue obtained from morbidly obese patients. METHODS: We used targeted microarrays representing human genes involved in the inflammatory and fibrogenic reactions to profile visceral adipose samples of 15 well-matched NASH patients with and without fibrosis. Additionally, visceral adipose samples were subjected to real-time polymerase chain reaction profiling of 84 inflammations related genes. RESULTS: Eight genes (CCL2, CCL4, CCL18, CCR1, IL10RB, IL15RA, and LTB) were differentially expressed in NASH with fibrosis. Additionally, an overlapping but distinct list of the differentially expressed genes were found in NASH with type II diabetes (DM; IL8, BLR1, IL2RA, CD40LG, IL1RN, IL15RA, and CCL4) as compared to NASH without DM. CONCLUSIONS: Inflammatory cytokines are differentially expressed in the adipose tissue of NASH with fibrosis, as well in NASH with DM. These findings point at the interaction of adipose inflammatory cytokines, DM, hepatic fibrosis in NASH, and its progression to cirrhosis and end-stage liver disease. PMID- 19280269 TI - Incidence of ceramic liner malseating in Trident acetabular shell. AB - The low wear rates associated with ceramic hip articulations have made them a popular bearing for younger patients. Although few complications have been observed, one report revealed several instances of incomplete seating of the ceramic liner in the metallic shell. We performed a cohort study of consecutive THAs using a ceramic-ceramic bearing. Radiographic analysis showed 50 (7.2%) of the group of 694 hips had evidence of incomplete seating of the liner in the metallic shell. Although we observed no adverse effects at 6 to 12 weeks, we encourage surgeons to carefully assess liner placement in the metal shell at the time of surgery to avoid this unintended consequence and to assess placement at the time of followup so patients can be properly followed when incomplete seating is identified. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19280270 TI - Focal breast uptake of 99m-Tc sestamibi in a hematoma. PMID- 19280272 TI - Prior treatment with vitamin K(2) significantly improves the efficacy of risedronate. AB - Prior 8-week treatment with menatetrenone, MK-4, followed by 8-week risedronate prevented the shortcomings of individual drugs and significantly increased the strength of ovariectomized ICR mouse femur compared to the ovariectomized (OVX) controls. Neither MK-4 following risedronate nor the concomitant administration may be recommended because they brought the least beneficial effect. INTRODUCTION: The objective of this study was to determine the best combinatory administration of risedronate at 0.25 mg/kg/day (R) with vitamin K(2) at approximately 100 microg MK-4/kg/day (K) to improve strength of osteoporotic mouse bone. METHODS: Thirteen-week-old ICR mice, ovariectomized at 9-week, were treated for 8 weeks with R, K, or R plus K (R/K), and then, either the treatment was withdrawn (WO) or switched to K or R in the case of R and K. After another 8 weeks, the mice were killed, and mechanical tests and analyses of femur properties by peripheral quantitative computed tomography, microfocus X-ray tube computed tomography, and confocal laser Raman microspectroscopy were carried out. RESULTS: The K to R femur turned out superior in parameters tested such as material properties, bone mineral density, BMC, trabecular structure, and geometry of the cortex. The increased cross-sectional moment of inertia, which occurred after K withdrawal, was prevented by risedronate in K to R. In addition to K to R, some properties of R to WO diaphysis and K to WO epiphysis were significantly better than OVX controls. CONCLUSION: Prior treatment with MK-4 followed by risedronate significantly increased femur strength in comparison to the OVX controls. PMID- 19280273 TI - Determinants of 25-hydroxyvitamin D levels in African-American and Caucasian male veterans. AB - SUMMARY: Among 307 males seen in VA Medical Center, independent determinants (p < 0.01 for all) of serum 25-hydroxyvitamin D [25(OH)D] levels included race, vitamin D supplements, BMI, dietary calcium intake and smoking, but not age. Negative association between 25(OH)D and parathyroid hormone (PTH) was similar for Caucasian and African-American men. INTRODUCTION: In this prospective cohort study, we examined determinants of serum 25-hydroxyvitamin D [25(OH)D] levels and the relationship between 25(OH)D and PTH levels and body mass index (BMI). METHODS: Male veterans (n = 307) were recruited at a VA Medical Center. Serum levels of PTH and 25(OH)D were obtained. Surveys and chart reviews were completed. Vitamin D insufficiency was defined as 25(OH)D <30 ng/ml. Univariate and multivariate regression analyses were performed. RESULTS: Among 232 African American (AA) men (mean +/- SD), 25(OH)D level (21.4 +/- 10.4 ng/ml) was lower and prevalence of insufficiency (80%) was higher than among 75 Caucasians (C; 28.5 +/- 11.1 ng/ml and 53%, respectively, p < 0.01 for both). In multivariate regression analysis, independent determinants (p < 0.01 for all) of 25(OH)D levels included AA race, vitamin D supplements, BMI, dietary calcium intake, and smoking. Despite lower 25(OH)D levels in African-Americans, PTH levels were similar to those seen in Caucasians. There was a significant (p < 0.02) negative linear association between 25(OH)D and PTH in African-American (r(2) = 0.05) and Caucasian (r(2) = 0.08) men, and there was no difference between the slopes of the relationship. CONCLUSIONS: 25(OH)D levels are determined by modifiable risk factors such as vitamin D supplementation in both AA and C males. The negative association between 25(OH)D and PTH is similar between the two races. PMID- 19280274 TI - Laparoscopic ventral hernia repair during pregnancy. AB - BACKGROUND: Laparoscopic ventral hernia repair in comparison to open herniorrhaphy results in reduced length of stay, less post-operative pain, earlier return to work, and reduced complications for the repair of complex ventral hernias. The laparoscopic approach has been the standard of care for complex or large ventral hernias for non-pregnant patients over the past decade. Despite evidence that demonstrates that laparoscopy is safe during pregnancy, there is currently no consensus regarding the indications, contraindications, patient selection and post-operative care of pregnant patients evaluated for laparoscopic ventral herniorrhaphy. METHODS: The medical records of our pregnant patient who underwent laparoscopic ventral herniorrhaphy were reviewed for demographics, operative indications, surgical technique, perioperative complications, recurrence, and outcome of the pregnancy. A Medline search using the terms: laparoscopy, surgery, and pregnancy was performed to review the literature from 1997 to 2007. RESULTS: This case report represents the first published description of a safe and successful laparoscopic approach to the repair of a complex ventral hernia in a woman at 21 weeks gestation. The discussion reviews the current literature regarding the safety of laparoscopy in pregnant women and highlights techniques to reduce perioperative morbidity and risk to the fetus. CONCLUSIONS: Laparoscopic ventral hernia repair can be safe during pregnancy with appropriate fetal monitoring and consideration of physiologic changes that occur during parturition. Elective procedures should be delayed until after delivery and all semi-elective surgeries until organogenesis is completed during the second trimester. PMID- 19280275 TI - Anastomotic leakage after low anterior resection for rectal cancer: facts, obscurity, and fiction. AB - The subject of anastomotic leakage after low anterior resection (LAR) for rectal cancer remains controversial. Risk factors have been discussed in several studies but the findings are often inconclusive. This review evaluates these studies and separates the known risk factors into those that are well documented, those that are obsolete, and those that require further research. We searched the Medline and PubMed databases using the keywords: "leakage," "low anterior resection," "rectal cancer," "risk factors," and their combinations. There were no language or publication year restrictions. References in published papers were also reviewed. Each risk factor was evaluated and discussed separately. The evidence suggests that low anastomoses are more prone to leakage. Other well-documented risk factors are male sex, smoking, and preoperative malnutrition. Routine mobilization of the splenic flexure and the use of a J-pouch seem to reduce the leakage rate. The effect of preoperative chemo-radiotherapy is under scrutiny. The indications for a protective stoma remain debatable. Omentoplasty, bowel preparation, the use of a drain, and tumor stage do not seem to affect the leakage rate. The type of operation (open or laparoscopic) and anastomosis (hand sewn or stapled) is not crucial. PMID- 19280276 TI - Persistent perineal sinus: incidence, pathogenesis, risk factors, and management. AB - This review discusses the incidence, pathogenesis, risk factors, diagnosis, and therapeutic options for persistent perineal sinus (PPS), defined as a perineal wound that remains unhealed more than 6 months after surgery. The incidence of PPS after surgery for inflammatory bowel disease (IBD) ranges from 3% to 70% and after abdominoperineal resection (APR) for low rectal cancer, it can be up to 30%. These unhealed wounds are frequently related to perioperative pelvic or perineal sepsis. Crohn's disease (CD) and neoadjuvant radiation therapy are also important risk factors. The management of PPS is based on an understanding of pathogenesis and clinical grounds. The advantages and disadvantages of the current therapeutic approaches, including the topical administration of various drugs, vacuum-assisted closure, and perineal reconstruction with a muscle flap or a myocutaneous flap are also discussed. PMID- 19280277 TI - Innate immune therapy with a Bacillus Calmette-Guerin cell wall skeleton after radical surgery for non-small cell lung cancer: a case-control study. AB - PURPOSE: We investigated whether adjuvant immunotherapy with Bacillus Calmette Guerin (BCG) cell wall skeleton (CWS) and surgical resection was better than resection, with or without other adjuvant therapy, for patients with non-small cell lung cancer (NSCLC). METHODS: The case group comprised 71 patients who underwent radical surgery for NSCLC, followed by BCG-CWS immunotherapy, with follow-up data available. The case-control study was designed with one control selected for each case-group patient. Each control was matched by pathological stage and year of birth (+/-5 years). BCG-CWS 200 microg was inoculated intracutaneously in the upper arm four times per week (sensitization phase); then at 4-week intervals (therapeutic phase). RESULTS: The case-group patients received 45 +/- 22.6 (average +/- SD) cycles of BCG-CWS inoculation. Overall 5 year and 10-year survival rates were 71% and 61% for the case-group patients, and 63% and 43% for the control-group patients. The survival rate of the case group was better than that of the control group (not significant; P = 0.114). The same trend was seen in the patients with stage III or N+ NSCLC (not significant; P = 0.114, P = 0.168). There were no life-threatening adverse events. CONCLUSIONS: BCG-CWS immunotherapy seemed to improve survival after resection of NSCLC, especially locally advanced NSCLC. Moreover, this immunotherapy did not compromise quality of life during treatment. PMID- 19280278 TI - The triangulating stapling technique for cervical esophagogastric anastomosis after esophagectomy. AB - PURPOSE: To evaluate the safety and feasibility of the triangulating stapling technique (TST) for cervical esophagogastric anastomosis after esophagectomy (CEGA). METHODS: The subjects were 123 patients who underwent transthoracic esophagectomy with three-field lymph node dissection and reconstruction with a 3.5-cm wide gastric tube, for thoracic esophageal cancer. We performed the TST for CEGA in 33 patients operated on after December, 2006 (TST group) and hand sewn anastomosis in 90 patients operated on between 2002 and 2006 (HSA group). RESULTS: In the TST group, CEGA was performed in an end-to-end fashion using three linear staplers. The first anastomosis was applied to the posterior walls of the remnant esophagus and gastric tube in an inverted fashion. The second and the third anastomoses were done in an everted fashion to make the anterior wall. The end-to-end HSA was performed with interrupted sutures using 4-0 absorbable material. Anastomotic leakage occurred in only 1 (3.0%) of the 33 TST patients, but in 13 (14.4%) of the 90 HSA patients (P = 0.07). The frequency of anastomotic stenosis was 9.1% and 25.6% in the TST and HSA groups, respectively (P < 0.05). CONCLUSIONS: Cervical esophagogastric anastomosis using TST may reduce the frequency of anastomotic leakage and stenosis. This technique is a safe and reliable alternative for CEGA after esophagectomy. PMID- 19280279 TI - D2 plus para-aortic lymphadenectomy versus standardized D2 lymphadenectomy in gastric cancer surgery. AB - PURPOSE: To evaluate the survival benefits and safety of D2 plus para-aortic lymphadenectomy (D2 + PALD) for gastric carcinoma. METHODS: Patients with gastric carcinoma, who agreed to undergo D2 + PALD between February 2001 and December 2003, were allocated to the D2 + PALD group, and compared with a control group who underwent D2 lymphadenectomy. Patients were followed up until August 2007. RESULTS: Sixty-two patients were allocated to the D2 + PALD group, and a concurrent 55 patients were allocated to the D2 group. The mean follow-up period was 57.6 (range 43.0-77.6) months, with 11.1% lost to follow-up. The morbidity and mortality rates were 24.2% and 0% in the D2 + PALD group, and 27.3% and 1.8% in the D2 group, respectively. The overall 3- and 5-year survival rates were 77.5% and 65.8% in the D2 + PALD group, and 73.2% and 66.1% in the D2 group, respectively, without a significant difference. The frequency of metastasis to the para-aortic lymph nodes (PALN) was 8.1%. The logistic regression revealed that PALN metastasis was correlated to metastasis of No. 8a and No. 9 lymph nodes (P = 0.021 and P = 0.030, respectively). CONCLUSION: Although D2 + PALD can be performed safely with an acceptable incidence of complications when performed by well-trained gastrointestinal surgeons, its survival benefits are not significantly greater than those of D2 lymphadenectomy. Therefore, routine D2 + PALD should not be recommended. PMID- 19280280 TI - Colonic tattooing using fluorescence imaging with light-emitting diode-activated indocyanine green: a feasibility study. AB - PURPOSE: We investigated the feasibility of a fluorescence imaging technique using light-emitting diode (LED)-activated indocyanine green (ICG) fluorescence. METHODS: Indocyanine green injections were given to patients undergoing preoperative colonoscopy for early colon cancer or colon adenoma. During subsequent laparotomy, the colon was first observed with the naked eye, and then using a prototype machine with a charge-coupled device (CCD) video camera equipped with a cutoff filter and a LED at a wavelength of 760 nm as the light source. RESULTS: LED-induced fluorescence showed tumor localization clearly and accurately in all ten patients (100%) enrolled in this study, whereas it was seen with the naked eye as a green spot in only two patients (20%) (P = 0.0077; Wilcoxon's signed-rank test). There were no complications of LED-induced fluorescence and no inflammatory signs were noted on the hematoxylin-eosin stained slides for the identified injection sites in the resected specimens. CONCLUSIONS: Colonic tattooing using this fluorescence imaging technique of LED activated ICG fluorescence is a new concept of colonic marking based on the characteristics that ICG is a near infrared fluorescent dye, and is useful, without any adverse effects, to identify perioperatively the tumor localization. PMID- 19280281 TI - Pylorus-preserving pancreaticoduodenectomy versus conventional pancreaticoduodenectomy for pancreatic adenocarcinoma. AB - PURPOSE: A few randomized controlled trials have questioned the justification of pylorus-preserving pancreaticoduodenectomy (PpPD) for pancreatic cancer and periampullary cancer. However, the characteristics of pancreatic cancer are remarkably different from those of other periampullary cancers, so the outcomes of PD and PpPD for pancreatic cancer are being re-evaluated. METHODS: We studied retrospectively, 55 patients who underwent PpPD at Wakayama Medical University Hospital between 1999 and 2005, when PpPD was available, for pancreatic head adenocarcinoma. The main outcome measures were the postoperative complications, mortality, and survival of the patients who underwent PpPD vs. those who underwent conventional pancreaticoduodenectomy (PD) for pancreatic adenocarcinoma. RESULTS: There were no significant differences between PD and PpPD in postoperative complications; however, the incidences of delayed gastric emptying (DGE) differed significantly according to the type of reconstruction (P < 0.01). The body weight ratio and the incidence of diarrhea 6 months after PpPD and PD were similar. Patients treated with PD had a higher duodenal invasion rate than those treated with PpPD (P < 0.05); therefore, the cause-specific survival of the PpPD patients was better than that of the PD patients (P < 0.05). CONCLUSION: The surgical outcomes and incidence of postoperative complications in this series suggest that PpPD is an appropriate surgical procedure for pancreatic adenocarcinoma. PMID- 19280282 TI - Early postoperative enteral nutrition is useful for recovering gastrointestinal motility and maintaining the nutritional status. AB - PURPOSE: The efficacy of enteral nutrition in postoperative nutritional management is known, but the effects on gastrointestinal motility and nutrition have not yet been elucidated. The purpose of this study was to compare the effects of enteral and parenteral nutrition soon after open abdominal surgery on gastrointestinal motility and nutritional status. METHODS: A partial resection of rectum models was prepared to compare two types of nutrient administration: enteral nutrition and total parenteral nutrition. The differences between the effects of nutrition types in terms of gastrointestinal motility and nutritional status were investigated. RESULTS: Enteral nutrition contributed to recovery of gastrointestinal motility and maintenance of nutritional status. CONCLUSION: Enteral nutrition should therefore be initiated soon after surgery if the gastrointestinal tract is available. PMID- 19280283 TI - Deep venous thrombosis caused by congenital absence of the inferior vena cava: report of a case. AB - Although anomalies of the inferior vena cava (IVC) are seen frequently in a clinical setting, congenital absence of the IVC (AIVC) is rare. However, anomalies of the IVC should be considered in young patients suffering from recurrent and idiopathic DVT. We report a case of DVT possibly caused by AIVC in a 27-year-old man, and discuss the clinical features, diagnosis, and treatment of this unusual entity. PMID- 19280284 TI - Signet ring cell carcinoma of the appendix manifesting as colonic obstruction and ovarian tumors: report of a case. AB - Appendiceal cancer is rare and associated with a poor prognosis because it is usually found at an advanced stage. We report a case of appendiceal adenocarcinoma manifesting as a colonic obstruction with a lower abdominal mass. Laparotomy revealed bilateral ovarian tumors and a small appendiceal tumor with peritoneal metastases. We performed ileocecal resection, colectomy, and oophorectomy, following which a histological diagnosis of signet ring cell carcinoma was made. Immunohistochemical analysis revealed positive expression of cytokeratin 7 and 20, and mucin core protein 2 (MUC2), compatible with appendiceal cancer and Kruckenberg metastases. When a patient is found to have disseminated pelvic signet ring cell carcinoma of unknown origin, the appendix should be considered as a possible primary site. PMID- 19280285 TI - Carcinosarcoma of the gallbladder producing alpha-fetoprotein and manifesting as leukocytosis with elevated serum granulocyte colony-stimulating factor: report of a case. AB - A 69-year-old man was referred to our hospital for investigation of leukocytosis and a persistent fever of 38 degrees C, but we could find no evidence of a specific infection. The leukocyte count was 18,000/mm(3), and the serum granulocyte colony-stimulating factor (G-CSF) and alpha-fetoprotein (AFP) levels were both elevated, at 66.3 pg/ml and 1,495 ng/ml, respectively. Computed tomography (CT) showed a gallbladder tumor and we performed extended cholecystectomy. Postoperatively, the fever subsided and the leukocyte count, serum G-CSF and AFP level normalized. Histologically, the tumor was a carcinosarcoma of the gallbladder. Immunohistochemical staining of the tumor cells was positive for AFP, but negative for G-CSF. This is the first report of a carcinosarcoma of the gallbladder producing AFP. The laboratory findings and clinical course strongly suggested that the tumor produced not only AFP, but also G-CSF. PMID- 19280286 TI - Intrahepatic cholangiocarcinoma arising 34 years after excision of a type IV-A congenital choledochal cyst: report of a case. AB - We report a rare case of intrahepatic cholangiocarcinoma (IHCC) arising many years after excision of a type IV-A congenital choledochal cyst. A 44-year-old man was transferred to our hospital with acute cholangitis more than 34 years after several operations for congenital biliary dilatation. Imaging showed a huge tumor in the left medial section of the liver, extending to the porta hepatis. Although he had no jaundice, the intrahepatic bile ducts showed cylinder-like dilatation with narrowing of the hilar bile duct. At surgery, the tumor was found to arise from the dilated intrahepatic bile duct just above the narrow portion. He underwent a left hepatic trisectionectomy with a vascular procedure. Microscopically, the tumor was confirmed to be moderate-to-well-differentiated tubular adenocarcinoma. Thus, when the narrow segment is left untouched, careful long-term follow-up is important to detect new lesions at an early stage. PMID- 19280287 TI - Laparoscopic cholecystectomy and common bile duct exploration for cholecystocholedocholithiasis with a left-sided gallbladder: report of a case. AB - A 75-year old woman was admitted to our hospital with right upper quadrant pain, vomiting, and jaundice. Laboratory findings showed elevated total bilirubin, alkaline phosphatase, gamma-glutamyl transpeptidase, and C-reactive protein levels. Abdominal ultrasonography (US) and drip infusion cholangiographic computed tomography (DIC-CT) showed not only cholecystocholedocholithiasis, but a gallbladder located left of the round ligament and close to the lateral segments of the liver. We performed laparoscopic cholecystectomy (LC) with choledocholithotomy for suspected cholecystocholedocholithiasis with a left-sided gallbladder. Routine ports were inserted in the American configuration for LC. The gallbladder was normogradely separated from the gallbladder fossa, and the fundus of the gallbladder was lifted ventrally and toward to the patient's right shoulder. These procedures provided the usual view for laparoscopic choledochotomy. The patient recovered uneventfully and was discharged on postoperative day 10. To our knowledge, this is the first report of laparoscopic common bile exploration in a patient with a left-sided gallbladder. PMID- 19280288 TI - Intrahepatic bile duct dilatation with a liver cyst and hemangioma: report of a case. AB - We report a case of intrahepatic bile duct dilatation with a liver cyst and hemangioma. A 58-year-old woman was referred for investigation of a cystic lesion and peripheral intrahepatic bile duct dilatation in the left lateral segment of the liver. Abdominal computed tomography (CT) and magnetic resonance imaging (MRI) showed dilatation of the intrahepatic bile ducts in the left lateral segment, near a 4.5-cm cystic lesion. Early enhancement into the delayed phase was seen in the dorsal part of the cystic lesion. Celiac angiogram showed a belt shaped hypervascular area, but no encasement or irregularity of the artery. To exclude malignancy, we performed a left lobectomy. Microscopically, the cyst was lined with a single layer of flattened epithelial cells and a spongy tumor was diagnosed as cavernous hemangioma, which compressed the bile duct. The histopathological diagnosis was biliary stenosis associated with cavernous hemangioma of the liver. Invasive surgery may be avoided by awareness of this unusual benign pathology. PMID- 19280289 TI - Gastric tube interposition for corrosive esophagitis associated with pyloric stenosis. AB - Corrosive esophagitis, caused by swallowing corrosive acid or alkali, results in cicatricial stricture of the esophagus. The stricture is often accompanied by pyloric stenosis because strong acids act synergistically with gastric juice. Resection of both the esophagus and stomach is usually necessary, and the colon or jejunum is used as an esophageal substitute. We describe how we successfully treated corrosive esophagitis associated with pyloric stenosis, by performing gastric tube interposition for the esophageal reconstruction. After resecting the injured distal part of the stomach, we pulled the pedunculated gastric tube up to the cervix after anastomosis to the jejunal limb in a Roux-en-Y fashion. This reconstruction procedure prevented excessive organ sacrifice and was minimally invasive. Thus, esophageal reconstruction by interposition using a pedunculated gastric tube can be used effectively to treat corrosive esophagitis associated with pyloric stenosis. PMID- 19280290 TI - Side-to-end anastomosis in a colostomy for acute malignant large-bowel obstruction: side-to-end anastomosis with a colostomy (STEC procedure). AB - This report describes the use of side-to-end anastomosis in a colostomy for an acute malignant large-bowel obstruction. A 59-year-old man presented with a colonic obstruction due to advanced descending colon cancer. The preoperative imaging studies revealed a complete obstruction of the descending colon at the site of the splenic flexure, a remarkably dilated transverse colon, and no other metastatic lesions. Side-to-end anastomosis was performed with the colostomy because of the high comorbidity associated with such cases. When the patient's general condition improved, a stoma closure was performed under local anesthesia. In conclusion, a side-to-end anastomosis with a colostomy (STEC procedure) was found to be a simple, useful, and cost-effective technique for an acute malignant large-bowel obstruction, particularly in a high-risk patient. PMID- 19280291 TI - Superior approach for the exclusion of hepatic veins in major liver resection: a safe and easy technique. AB - We describe a technique for isolating and excluding the hepatic veins during liver resection. First, the bare area near the right and left wall of the suprahepatic inferior vena cava (IVC) is dissected, exposing the right, left, and superior walls of the right hepatic vein (RHV) and the left-middle hepatic vein (LMHV). Two Satinsky clamps are used to clamp the roots of the right and common trunk of the LMHV, parallel to the IVC. It is not necessary to dissect the posterior wall of the hepatic veins. We used this method during major liver resection in 65 patients. The mean dissecting time of each hepatic vein was 7.31 +/- 3.6 min. No hepatic vein was lacerated during dissection and exclusion. The postoperative complication rate was 31.2%. Thus, the superior approach is a safe and easy maneuver when the posterior wall of the hepatic vein is difficult to dissect due to tumor invasion. PMID- 19280292 TI - In vitro antifungal combination effects of micafungin with fluconazole, voriconazole, amphotericin B, and flucytosine against clinical isolates of Candida species. AB - Micafungin (MCFG) is an echinocandin antifungal agent that exhibits potent activity against most species of Candida and Aspergillus. We investigated the in vitro antifungal combination effects of MCFG with four other antifungal agents - fluconazole (FLCZ), voriconazole (VRCZ), amphotericin B, and flucytosine - against clinical isolates of 54 Candida spp. by checkerboard analysis. The synergistic antifungal effects of MCFG-FLCZ and MCFG-VRCZ were 11% and 15%, respectively, and the latter displayed a synergistic activity of 63% against Candida glabrata. Antagonism was not observed in any of the combinations tested. PMID- 19280293 TI - Molecular characterizations of carbapenem and ciprofloxacin resistance in clinical isolates of Pseudomonas putida. AB - To analyze the genetic mechanisms of carbapenem and ciprofloxacin resistance in clinical isolates of Pseudomonas putida, 27 clinical isolates (comprising 11 carbapenem- and ciprofloxacin-resistant strains, 13 carbapenem-resistant and ciprofloxacin-susceptible strains, and 3 carbapenem- and ciprofloxacin susceptible strains) were collected from different patients. Carbapenem resistance was examined by polymerase chain reaction (PCR) and DNA sequencing for metallo-beta-lactamase (MBL) and integrase genes (IntI-1 and IntI-3), and by reverse transcriptase-PCR (RT-PCR) for expression of the porin gene (oprD). Ciprofloxacin resistance was characterized by PCR and DNA sequencing for mutations in the quinoloneresistance determining regions of the gyrA and parC genes. The blaIMP-1 MBL and intI-1 and/or intI-3 genes were detected in all carbapenem-resistant strains, and decreased expression of the oprD gene as compared to carbapenemsusceptible strains was observed in several strains. All the 11 strains with ciprofloxacin minimal inhibitory concentrations (MICs) of > or =64 mg/l had substitution in GyrA (Thr83Ile), and one (ciprofloxacin MIC of 512 mg/l) of these strains also had substitution in ParC (Ser87Leu). Overproduction of the efflux pump was observed in 10 of the 11 ciprofloxacin resistant strains. We concluded that the production of IMP-1 type MBL was the most critical factor in developing high-level resistance to carbapenems, and mutations in the target proteins and overproduction of the efflux pump synergistically contribute to the acquisition of high-level resistance to ciprofloxacin in clinical isolates of P. putida. PMID- 19280294 TI - Monte Carlo simulation for evaluation of the efficacy of carbapenems and new quinolones against ESBL-producing Escherichia coli. AB - Extended-spectrum beta-lactamase (ESBL)-producing bacteria are known to be resistant to penicillins, cephalosporins, and monobactams because of their substrate specificity, and these bacteria are sensitive only to a narrow range of antimicrobial agents. The present study was undertaken to evaluate the efficacy of carbapenems and the new quinolones against ESBL-producing Escherichia coli, using a Monte Carlo simulation based on the pharmacokinetic/pharmacodynamic (PK/PD) theory. The time above MIC (TAM, %) served as the PK/PD parameter for carbapenems, with the target level set at 40%. The AUC/MIC served as the PK/PD parameter for the new quinolones, with the target level set at more than 125. In the analysis of drug sensitivity, the MIC50 of all carbapenems other than imipenem was low (0.03 microg/ml), while the MIC50 of the new quinolones was higher (1-2 microg/ml). The probability of achieving the PK/PD target with carba penems after two doses at the usual dose level, as determined by the Monte Carlo simulation, was high for each of the carbapenems tested (99.0% for biapenem, 99.60% for meropenem, and 95.03% for doripenem), except for imipenem. Among the new quinolones, the highest probability of achieving the PK/PD target was obtained with pazufloxacin (42.90%). Thus, the results of the present study have revealed that carbapenems are effective at the regular dose and can be used as the first-choice antibiotics for ESBL-producing E. coli because the resistance ratios for carbapenems are low compared to those of the new quinolones. PMID- 19280295 TI - Clinical investigation of isolated bacteria from urinary tracts of hospitalized patients and their susceptibilities to antibiotics. AB - Complicated urinary tract infections (UTIs) are often difficult to treat, partly because of the existence of isolated antibiotic-resistant strains. Even though the definition of UTI is determined by the quantity of cultured bacteria, it has been unclear if the quantity of cultured UTI bacteria affects their susceptibility to antimicrobial agents. Also, the gram stain is generally performed to assume the causative bacteria and their quantity before culture results can be obtained; therefore, we could start to use effective antibiotics if the relationship between bacterial quantity and resistance to antimicrobial agents were clear. We studied patients diagnosed as having complicated UTIs at the Urological Department in Kobe University Hospital between June 2002 and March 2003 and analyzed the relationships between the quantity of cultured bacteria and their antimicrobial MICs for antibiotics. The most commonly isolated bacteria were Serratia marcescens, Pseudomonas aeruginosa, Enterococcus faecalis, Escherichia coli, Staphylococcus epidermidis, Stenotrophomonas maltophilia, and methicillin-resistant Staphylococcus aureus. There was no significant correlation between the quantity of cultured bacteria and antimicrobial MICs in all the bacteria and antibiotics that we tested, suggesting that resistant phenotype, but not inoculum of the organism, did determine resistance to antibiotics. In conclusion, our investigation suggested the total number of isolated bacteria in urine culture did not determine the MICs and that inoculum of the bacteria might be important for this determination. PMID- 19280296 TI - Potential clinical benefit of the in situ hybridization method for the diagnosis of sepsis. AB - Despite the significant development of antibiotics, sepsis is still associated with high morbidity and mortality rates. The identification of pathologic organisms at an early stage of sepsis is critical to improve the outcome, but this is difficult to achieve with the conventional method of blood culture (BC). It has been demonstrated that the genes of pathogenic organisms surviving in neutrophils were detectable with in situ hybridization (ISH) and this method was useful for the accurate and rapid diagnosis of sepsis. In this study, we applied ISH to blood smears 60 patients with suspected sepsis. BC was also carried out using the same blood samples to investigate the diagnostic value of ISH. The number of positive results obtained by ISH was approximately four times higher than that obtained by BC (ISH, 25 [41.7%]; BC, 7 [11.7%]). The positive rate in the 21 patients given antibiotics was 61.9% by ISH (13 patients) and 4.7% by BC (1 patient). The antibiotic treatments targeting the organisms detected by either procedure showed a beneficial clinical outcome. Positive results by ISH were obtained earlier than those with BC (ISH, within 1 day; BC, several days). We conclude that ISH is a useful method for the rapid diagnosis of sepsis. PMID- 19280297 TI - Plasmodium vivax PCR genotyping of the first malaria case imported from South Korea into Japan. AB - Although indigenous malaria was successfully eradicated in Japan in 1959, malaria remains one of the most important health concerns in the control of imported infectious diseases. However, in South Korea, the re-emergence of indigenous vivax malaria was reported in 1993 in the Demilitarized Zone (the border region with North Korea), from where a vivax malaria case was imported into Japan in 2002. In this study, we conducted genotyping of the circumsporozoite protein gene, the apical membrane antigen-1 gene, and the merozoite surface protein-1 gene of Plasmodium vivax in one patient, and estimated the geographical origin of the parasites. This estimate was based on the findings of previous studies, which showed the coexistence of at least two distinct genotypes of antigenic molecules of endemic P. vivax in South Korea. One genotype is similar to that of a Chinese strain CH-5, and the other is similar to that of a North Korean isolate. The results of this study showed that the DNA sequences of the patient's P. vivax parasites were similar to those of the North Korean isolate. It may even be possible in the near future for seasonally synchronized North Korean P. vivax parasites to be imported into parts of Japan and to establish breeding populations. PMID- 19280298 TI - Systemic use of antibiotics does not prevent postoperative infection in elective colorectal surgery: a randomized controlled trial. AB - We assessed the clinical impact of the systemic use of antibiotics on postoperative infection in colorectal surgery. Perioperative administration prevents postoperative infection: a statement which is based on the results of five randomized controlled trials performed in the 1970s and 1980s. Our study design was a randomized controlled trial. We created two groups, one using the systemic antibiotic cefotiam (CTM), and the other using no antibiotic as the control. The primary end point was the overall postoperative infection rate. There were 100 patients assigned to this study. The patients were divided into two groups; the control group consisted of 51 cases and the CTM group had 49 cases. The backgrounds of the patients in the two groups were not significantly different. The overall postoperative infection rate was 28/51 (54.9%) in the control group and 25/49 (51.0%) in the CTM group. The surgical site infection (SSIs) (superficial, deep, and space/organ ) were 23/51 (45.1%) in the control group and 20/49 (40.8%) in the CTM group. No significant difference was observed between the CTM group and the control group regarding postoperative infection after elective colorectal surgery. PMID- 19280299 TI - A patient with acute myeloid leukemia who developed fatal pneumonia caused by carbapenem-resistant Bacillus cereus. AB - Bacillus cereus is known as a serious bacterial pathogen in neutropenic patients. B. cereus is often resistant to beta-lactams, including penicillins and cephalosporins. We report a case of fatal pneumonia caused by B. cereus in a patient with newly diagnosed acute myeloid leukemia (AML) during remission induction therapy. Cefepime was started for febrile neutropenia (FN) initially and was switched to panipem/betamipron, when fulminant pneumonia supervened. The isolated strain was resistant not only to cefepime but also to panipenem/betamipron. This is the first report of fulminant infection caused by carbapenem-resistant B. cereus in a neutropenic patient. B. cereus should be kept in mind as a target of empirical treatment when neutropenic patients develop pneumonia. PMID- 19280300 TI - Breakthrough disseminated zygomycosis induced massive gastrointestinal bleeding in a patient with acute myeloid leukemia receiving micafungin. AB - A 69-year-old man, who had been receiving prednisolone for 11 months for treatment of interstitial pneumonia, was diagnosed with acute myeloid leukemia. During induction therapy, he developed severe pneumonia. Although meropenem and micafungin were started, he died of circulatory failure owing to massive gastrointestinal bleeding. Autopsy specimens obtained from the stomach revealed fungal hyphae, which had invaded diffusely into submucosal vessels and caused the massive gastric bleeding. The same hyphae were also observed in both lungs. A diagnosis of disseminated zygomycosis was confirmed by its characteristic histopathological findings. Because zygomycetes are spontaneously resistant to the newer antifungal agents, such as voriconazole or micafungin, it seems likely that the prevalence of zygomycosis as a breakthrough infection may increase in the future. Zygomycosis is a rare, but life-threatening, deep fungal infection that appears in immunologically or metabolically compromised hosts. Its manifestations are clinically similar to those of invasive aspergillosis. In addition to the well-established epidemiology of zygomycosis, this case suggests the following new characteristics. (1) Although the gastrointestinal manifestation of zygomycosis is relatively rare, it is observed more frequently than invasive aspergillosis. (2) Gastrointestinal zygomycosis occasionally leads to the development of necrotic ulcers and may induce hemorrhagic shock.(3) We should be cautious of an occurrence of breakthrough zygomycosis when we use echinocandins for patients with known risk factors, especially steroid use and neutropenia. (4) For patients who are receiving broad-spectrum antibiotics and echinocandins, who are negative for culture studies and aspergillus antigen, and who present with unresolved fever, it is important to make a prompt clinical diagnosis of zygomycosis. PMID- 19280301 TI - Mycobacterium abscessus from respiratory isolates: activities of drug combinations. AB - Mycobacterium abscessus is the most resistant of rapidly growing pathogenic mycobacteria. We performed a screening of the in vitro sterilizing capacity of various antibiotic combinations against two strains isolated from respiratory samples (in vitro activity of each antibiotic: 51 combinations of two drugs and 47 of three drugs). Our findings confirm that clarithromycin is the most useful drug. The combination of this drug with linezolid exhibits good activity. In addition, the fluoroquinolones and rifabutin in association with clarithromycin may have clinical utility because they are administered orally. Therapeutic alternatives in this type of infections should be evaluated in greater depth. PMID- 19280302 TI - Delayed insertion of blood culture bottles into automated continuously monitoring blood culture systems increases the time from blood sample collection to the detection of microorganisms in bacteremic patients. AB - This study examined the effects on patients with bacteremia of delaying the insertion of a blood culture bottle into an automated, continuously monitoring blood culture system. We investigated the time taken from the collection of blood samples (collection) to the insertion of blood culture bottles inoculated with blood samples into the instrument (insertion), and compared the mean detection time from collection to a positive signal from the instrument with the time between collection and insertion. The study was conducted from January 2003 to December 2004 at Kyoto University Hospital. Insertion into the system on the day of blood sample collection was defined as same-day insertion, and insertion on a different day to collection was defined as delayed insertion. The 7394 aerobic and anaerobic blood culture bottle sets obtained during the study period included 4361 sets with same-day insertion and 3033 sets with delayed insertion. For same day insertion, 458 microorganisms were isolated from 432 positive sets in which at least one blood culture bottle was positive. For delayed insertion, 405 microorganisms were isolated from 379 positive sets in which at least one blood culture bottle was positive. The mean detection time for all microorganisms was significantly earlier for same-day insertion than for delayed insertion (28.3 h vs. 45.0 h, respectively, P < 0.0001). Delays from collection to insertion affect the time from collection to the detection of microorganisms. PMID- 19280303 TI - Superior in vitro activity of carbapenems over anti-methicillin-resistant Staphylococcus aureus (MRSA) and some related antimicrobial agents for community acquired MRSA but not for hospital-acquired MRSA. AB - Eighty-eight strains of Panton-Valentine leukocidin (PVL)-positive and -negative community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) and 152 strains of hospital-acquired MRSA (HA-MRSA) were examined for susceptibility to carbapenems, oxacillin, and other antimicrobial agents. CA-MRSA strains were more susceptible to carbapenems (MIC(90), 1-4 microg/ml) than HA-MRSA strains (MIC(90), 32-64 microg/ml). Among the carbapenems examined, CA-MRSA strains were most susceptible to imipenem (MIC(50), 0.12 microg/ml; MIC(90), 1 microg/ml). A similar tendency was observed with oxacillin, but less markedly (MIC(90): 32 microg/ml for CA-MRSA and > or =256 microg/ml for HA-MRSA). This difference was also observed between CA-MRSA and HA-MRSA in susceptibility levels to cephems, erythromycin, clindamycin, and levofloxacin, but not to ampicillin, vancomycin, teicoplanin, linezolid, and arbekacin. The data indicate that, in terms of MIC(50) or MIC(90) values, CA-MRSA is 64-256 times more susceptible to imipenem than HA-MRSA, and for CA-MRSA, some carbapenems, e.g., imipenem, show better in vitro activity than anti-MRSA or some related agents. PMID- 19280304 TI - Procalcitonin and its utility as a diagnostic and prognostic assay in patients with renal dysfunction and immunosuppressive conditions. PMID- 19280306 TI - Surgery of the thymus in Japan. AB - The thymus has long been an organ of mystery. Today, it is known to be central to the construction of our immune system. I focus on the days when no one was sure what the thymus's function was. I review the Japanese literature regarding the beginnings of surgery of the thymus, especially the surgery for thymoma and thymectomy, which was performed as a treatment for myasthenia gravis. PMID- 19280307 TI - Rationale for video-assisted radical esophagectomy. AB - Despite many positive reports on video-assisted esophagectomy (VAE) for malignant esophageal tumors, VAE remains a lesser used procedure in Japan because of its technical difficulty and the strong desire of esophageal surgeons for thorough lymphadenectomy. In this article, we review former reports as well as our own experiences to demonstrate the feasibility and rationale for radical VAE as a standard operation for esophageal cancer. Although the reduction of surgical stress is not clear, it has been reported that VAE is associated with lower morbidity related to postoperative pulmonary complications, and we experienced a shorter period of postoperative systemic inflammatory response syndrome. VAE is advantageous not only because it is less invasive but also because of the possibility of a more meticulous operation through the magnifying effect of the surgical field and the alteration of the viewpoint of the surgeon. The number of dissected lymph nodes, a possible indicator of the extensiveness and thoroughness of lymphadenectomy, is reported to be the same for VAE as for conventional thoracotomy by many surgeons, and our experience was no exception. Although it is too early to discuss the prognostic effect, no obvious disadvantage in prognosis has been reported. We are convinced that radical VAE is not inferior in radicality as an operation for esophageal cancer. As the evaluation of radicality and safety of radical VAE differs somewhat from surgeon to surgeon, different attitudes about the indications for radical VAE remain. We cautiously suggest that use of this new procedure should move forward. PMID- 19280308 TI - Effect of low-dose landiolol, an ultrashort-acting beta-blocker, on postoperative atrial fibrillation after CABG surgery. AB - OBJECTIVE: Postoperative atrial fibrillation is the most common complication after coronary artery bypass grafting (CABG). This complication is associated with prolongation of the intensive care unit stay and hospitalization time with attendant increased hospitalization costs, and it is an important risk factor for perioperative cerebrovascular accidents. Landiolol is a newly developed ultrashort-acting beta-adrenoceptor antagonist with a half-life of 3 min that is eight times more cardioselective than esmolol. The purpose of this study was to investigate the prophylactic effect of continuous administration of low-dose landiolol on postoperative atrial fibrillation. METHODS: We reviewed all patients who underwent CABG alone at our hospital from April 2002 and September 2006. Patients with a previous history of atrial arrhythmias were excluded. The remaining patients were divided to two groups: landiolol group (n = 20), and control group (n = 35). Administration of landiolol started after cardiopulmonary bypass with a loading dose of 1.5-2.5 mug.kg(t-1).min(-1) and continued for the first 2 days after surgery. The incidence of postoperative atrial fibrillation was noted. Continuous variables were compared between groups by means of Student's t-test. Categorical variables were compared by means of the chi(2) test or Fisher's exact test. RESULTS: The occurrence of atrial fibrillation after CABG in the control group was statistically more than in the landiolol group (P = 0.04). There were no statistical differences between the groups regarding the cardiac index or the dose of inotropic agents during the perioperative period. CONCLUSION: Intraoperative and perioperative administration of low-dose landiolol has a preventive effect on the appearance of atrial fibrillation after CABG surgery. PMID- 19280309 TI - Histoculture drug response assay for gefitinib in non-small-cell lung cancer. AB - OBJECTIVE: There are many predictive factors for gefitinib sensitivity, including epidermal growth factor receptor (EGFR) gene mutation, EGFR copy number, and k ras mutation. To investigate all of them is too expensive. We evaluated the chemosensitivity for gefitinib in non-small-cell lung cancer (NSCLC) using a histoculture drug response assay (HDRA). METHODS: Surgically resected fresh tumor specimens from 22 patients with NSCLC were used. There were 13 male and 9 female patients, ranging in age from 49 to 84 (average 70) years old. Sixteen patients (73%) were smokers. Sixteen adenocarcinomas, four squamous cell carcinomas, and two other histological types were included. Small pieces of viable cancer tissue were placed on the collagen gel and then cultured for 7 days in the presence of gefitinib. RESULTS: The HDRA was successful in all specimens. A dose-response relation was observed between inhibition rates and gefitinib concentration (p = 0.016). The inhibition rate at 20 microg/ml (IR(20)) in adenocarcinoma without smoking (39.2% +/- 35.1%, n = 6) was higher than that with smoking (2.2% +/- 5.0%, n = 10, P = 0.001) and that of nonadenocarcinoma (16.9% +/- 23.6%, n = 6, P = 0.09). Gene mutation analysis was performed in two of three adenocarcinomas without smoking, which showed especially high IR(20) values, and sensitizing mutations were observed in these specimens. A cutoff inhibition rate of approximately 40%-50% appeared to be suitable for a concentration of 20 microg/ml. CONCLUSION: HDRA appears to be applicable for evaluating sensitivity to gefitinib in NSCLC. It provides a convenient method for predicting the response to gefitinib in patients with NSCLC whose fresh tumor specimens are available. PMID- 19280310 TI - Advantage of vein grafts for anomalous origin of a right coronary artery. AB - A 66-year-old man with anomalous origin of the right coronary artery suffered from chest pain. The results of coronary angiography and multidetector computer tomography indicated that the proximal right coronary artery was intermittently compressed, causing the ischemia. Coronary artery bypass grafting was regarded as a reliable treatment compared with percutaneous coronary intervention or other surgeries. Because of plentiful flow of the right coronary artery, we decided to use a vein graft to avoid competitive flow. Postoperative coronary angiography revealed intact flow in both the native coronary artery and the vein graft 1 year after the surgery. The myocardial ischemia seen on scintigraphy and the chest pain had disappeared. PMID- 19280311 TI - Traumatic ventricular septal defect following a stab wound to the chest. AB - A 51-year-old man who had been suffering from depression stabbed himself in the chest with an ice pick. At presentation, an ice pick lodged in the left fifth intercostal space was moving synchronously with his heartbeat. Echocardiography revealed that the tip was penetrating the anterior wall of the right ventricle. Because the patient was tamponading, an emergency operation was carried out. The ice pick was removed following the establishment of a cardiopulmonary bypass and pericardiotomy. The perforation of the right ventricle was closed with a pledget reinforced mattress stitch. On postoperative day 12, a holosystolic murmur was detected on auscultation. Transthoracic echocardiography revealed a ventricular septal defect 5 mm in diameter located near the apex. The pulmonary-tosystemic flow ratio was 1.1 by echocardiographic measurement. No sign of heart failure was present. Although it was agreed to manage the ventricular septal defect conservatively, careful echocardiographic follow-up is mandatory. PMID- 19280312 TI - Rare type of congenital aneurysm of the right sinus of Valsalva protruding superiorly into the pericardial space. AB - We report a rare type of aneurysm of the sinus of Valsalva. The orifice of the aneurysm was in the right sinus of Valsalva and protruded superiorly, which is a direction not typical of right sinus of Valsalva aneurysms. Intraoperatively, it became clear that the aneurysm was located in the right sinus of Valsalva with extracardiac protrusion. Inspection through the aortotomy revealed that the orifice of the aneurysm opened between the orifice of the right coronary artery and the commissure between the right and left coronary cusps. The aneurysm was obliterated with a pledgeted suture from outside the orifice of the aneurysm. Early prophylactic surgical treatment of the aneurysm makes the operation simple and prevents subsequent development of complications, such as compression of the coronary artery, thrombosis, and/or spontaneous rupture. PMID- 19280313 TI - Prosthetic mitral valve leaflets jammed by a balloon catheter. AB - We encountered a case in which prosthetic mitral valve leaflets were jammed by a transmitral balloon catheter during mechanical mitral valve replacement. The catheter, which was inserted in the left ventricle to render the leaflets incompetent during de-airing, forced the leaflets into the closed position. The leaflet edges pinched the catheter lumen and obstructed balloon deflation. Excessive inflation of the balloon and extraction of the catheter before complete deflation of the balloon should be avoided to prevent a potentially serious complication. PMID- 19280314 TI - Cardiac hemangioma presenting with neurological manifestations. AB - Primary cardiac tumors are rare. Hemangiomas represent about 5%-10% of benign cardiac tumors. Hemangiomas can be found in any heart chamber, but atria have been more frequently involved. Ventricular involvement is uncommon. Hemangiomas may have various presentations, but there have been few reports about neurological manifestations of such tumors in the English-language literature. We present a patient with a left ventricular hemangioma who presented with leftsided weakness and sudden onset of slurred speech. PMID- 19280315 TI - Giant primary liposarcoma of the chest. AB - Liposarcomas are the most common soft tissue sarcomas in adults, although liposarcomas of the chest are uncommon. We report two cases of giant liposarcoma in the mediastinum and chest wall, respectively. An 82-year-old man presented with a mass in the right upper mediastinum, as seen by computed tomography (CT). He had a past history of subcutaneous lipoma resection on his back (19 years previously). The patient underwent tumor resection with a right thoracotomy. A 58 year-old woman presented with an enlarging mass of the right lateral chest, involving the diaphragm and ribs, as seen by CT. She had a past history of subcutaneous lipoma resection of the right chest (18 years previously). The patient underwent an en bloc resection that included the tumor and a part of the right diaphragm and ribs. Histological examination of both patients' tumors revealed a well-differentiated liposarcoma, with no pathological relation to the previous lipoma resected in either case. PMID- 19280316 TI - Bronchial arteriovenous malformation with large aneurysm, resected by video assisted thoracic surgery. AB - Bronchial arteriovenous malformation with a large aneurysm appearing as a mass shadow in the right hilum on chest imaging was successfully extirpated thoracoscopically without any lung resection. Preoperative angiography showed tortuous, enlarged right bronchial arteries with an aneurysm connecting to the pulmonary artery, which could not be totally embolized via the transcatheter approach because of the length of the abnormal bronchial artery and possibility of embolotherapy-induced pulmonary infarction. Histological examination revealed defects of the media and internal elastic lamina of the resected bronchial arteries and aneurysm. The patient was free of hemoptysis and other airway symptoms 4 years after surgery. PMID- 19280317 TI - Mesenchymal cystic hamartoma of the lung: a rare cause of pneumothorax. AB - A 14-year-old girl had a left spontaneous pneumothorax. Because closed chest tube drainage failed to obtain expansion of the lung, thoracoscopic bullectomy was performed. Pathological examination of the bulla resulted in the diagnosis of mesenchymal cystic hamartoma, which is a rare cause of pneumothorax. PMID- 19280318 TI - Aspergilloma of the lung due to aspiration during nasal tube feeding. AB - An aspergilloma usually arises in a preexisting cavity in the lungs. We report a case of a young woman with anorexia nervosa who developed an aspergilloma some time after she suffered pneumonia due to aspiration during nasal tube feeding. PMID- 19280320 TI - Molecular characterization of adenoviral infections in Cuba: report of an unusual association of species D adenoviruses with different clinical syndromes. AB - Adenoviruses are common pathogens that are responsible for a wide variety of infectious syndromes. The objectives of this study were to identify and characterize members of different adenovirus species at the molecular level and to describe the correlation between viruses and clinical syndromes during a period of 4 years. Between 2002 and 2006, 45 of 512 respiratory specimens (8%) from patients with acute respiratory tract infection tested positive for adenovirus. Four adenovirus isolates from samples sent for enterovirus isolation were also analyzed. This research identified 49 confirmed cases of human adenovirus infection by PCR and/or viral culture. The most common diagnosis was upper respiratory infection (44%). Human adenovirus D was the major species found (59%), followed by Human adenovirus C (36%) and Human adenovirus B (4%). Human adenovirus 5 was the major serotype found producing bronchiolitis, followed by human adenovirus 6. In patients with upper respiratory infection, the major serotype found was human adenovirus 17. Viruses of the species Human adenovirus D were identified in seven (77%) cases of acute febrile syndrome. Four isolates from clinical materials obtained from patients with encephalitis, acute flaccid paralysis and meningoencephalitis were identified as belonging to the species Human adenovirus D. Our data demonstrate a surprising result about the identification of an unusual association of viruses of the species Human adenovirus D with different clinical syndromes. This observation could be evaluated as a possible indicator of the emergence of a novel strain but further studies are required. PMID- 19280321 TI - An autochthonous case of hepatitis C virus genotype 5a in Brazil: phylogenetic analysis. AB - Genotype 5 of hepatitis C virus (HCV) has been rarely identified in South America. A female of African descent who never left Brazil was found to be infected by this genotype in Mato Grosso state, Central Brazil. The patient denied drug injections and revealed that she had received blood transfusions several years before. One of her blood donors was identified and tested negative for anti-HCV and HCV RNA, as were her husband and offspring. Phylogenetic analysis of the E1 and NS5B regions confirmed that this HCV strain belonged to genotype 5a. However, the E1 region analysis indicates that our strain is not closely related to any sequences of genotype 5a from other geographical areas, diverging from the African and European subclades known so far. These data suggest that genotype 5a HCV might have been circulating at a low level in Brazil longer than previously supposed. PMID- 19280322 TI - Safety and robustness of coated pellets: self-healing film properties and storage stability. AB - PURPOSE: Aim of the study was to verify the safety of chlorpheniramine maleate pellets, coated with blends of poly(vinyl acetate) and poly(vinyl alcohol) poly(ethylene glycol) graft copolymer. Therefore, the impact of mechanical forces and storage conditions on the drug release was investigated. RESULTS: Similar release profiles before and after compression of the pellets to tablets underlined the high film robustness. A damage of the film coat with a razor blade resulted in a premature release, but without a burst. After a similar damage with a needle, the release profile remained almost unchanged, which indicated a swelling based self repair mechanism of the film. Additional studies were dedicated to the storage stability at three different conditions. A slightly delayed release was obtained after 6 months storage at 25 degrees C and a marginally accelerated release was measured after storage at elevated temperatures. No drug migration into the coating layer was detected during storage by confocal Raman microscopy. (1)H-NMR analysis during storage demonstrated, that no polymer or drug degradation had occurred and the plasticizer concentration remained constant. CONCLUSION: The polyvinyl based coating blend for modified release pellets demonstrated a high safety, due to their high robustness and compressibility as well as their satisfying storage stability. PMID- 19280323 TI - The impact of dose and solubility of additives on the release from HPMC matrix tablets--identifying critical conditions. AB - PURPOSE: The dissolution of HPMC matrix tablets containing different amounts of highly soluble (mannitol) or poorly soluble (dicalcium phosphate, DCP) was studied to deduce the parameters critical to release robustness. METHODS: The release of HPMC and additives was studied using a modified USP II method at two paddle stirring rates, 50 and 125 rpm, at HPMC content varying from 15% to 100%. RESULTS: At HPMC contents between 30% and 35% a critical point was identified and found crucial to the release from the HPMC/mannitol tablets. Below this point the matrix rapidly disintegrated in a non robust manner. At higher HPMC contents the mannitol release became increasingly diffusion controlled with maintained matrix integrity. The release robustness was lower for HPMC/DCP than HPMC/mannitol tablets at high HPMC contents, however, lacking critical points. The critical point was interpreted as the percolation threshold for HPMC and differences explained in terms of water transport into the matrix. CONCLUSION: The release robustness was lower for formulations with additives of low solubility having an erosion controlled release than for additives with higher solubility and a diffusion controlled release. However, for additives creating a steep osmotic pressure gradient, an HPMC content above the percolation threshold becomes vital for maintaining the release robustness. PMID- 19280324 TI - Intestinal lymphatic transport enhances the post-prandial oral bioavailability of a novel cannabinoid receptor agonist via avoidance of first-pass metabolism. AB - PURPOSE: To examine the effect of food on the oral bioavailability of a highly lipophilic, cannabinoid receptor agonist (CRA13) and to explore the basis for the food effect in lymph-cannulated and non-cannulated dogs. METHODS: Oral bioavailability was assessed in fasted and fed human volunteers and in lymph cannulated dogs. In fasted dogs, the extent of absorption and oral bioavailability was also examined following administration of radiolabelled CRA13. RESULTS: Food had a substantial positive effect on the oral bioavailability of CRA13 in human volunteers (4.3-4.9 fold increase in AUC(0 - infinity)) and in dogs. The absolute bioavailability of parent drug was low in fasted dogs (8-20%), in spite of good absorption (72-75% of radiolabelled CRA13 recovered in the systemic circulation). In post-prandial lymph-cannulated dogs, bioavailability increased to 47.5% and the majority (43.7%) of the dose was absorbed via the intestinal lymphatic system. CONCLUSIONS: The positive food effect for CRA13 does not appear to result from increased post-prandial absorption. Rather these data provide one of the first examples of a significant increase in bioavailability for a highly lipophilic drug, which is stimulated via almost complete post-prandial transport into the lymph, in turn resulting in a reduction in first-pass metabolism. PMID- 19280325 TI - Dendritic cell interactions with NK cells from different tissues. AB - INTRODUCTION: In recent years, it has been realized that innate lymphocytes do not act in isolation but potentiate their efficiency by interacting with each other, resulting even in the regulation of adaptive immune response. One such cross-talk exists between dendritic cells (DCs) and natural killer (NK) cells. Here, we summarize recent studies on which subsets of these two innate immune components participate in this interaction, how it influences immune responses, and to which extent similar stimuli are integrated by DCs and NK cells during innate immunity. CONCLUSION: We suggest that this cross-talk should be harnessed by activating both of these innate leucocyte populations with new adjuvant formulations for immunotherapies. PMID- 19280326 TI - A reexamination of correlations of amino acids with particular secondary structures. AB - Using the data from Protein Data Bank the correlations of primary and secondary structures of proteins were analyzed. The correlation values of the amino acids and the eight secondary structure types were calculated, where the position of the amino acid and the position in sequence with the particular secondary structure differ at most 25. The diagrams describing these results indicate that correlations are significant at distances between -9 and 10. The results show that the substituents on Cbeta or Cgamma atoms of amino acid play major role in their preference for particular secondary structure at the same position in the sequence, while the polarity of amino acid has significant influence on alpha helices and strands at some distance in the sequence. The diagrams corresponding to polar amino acids are noticeably asymmetric. The diagrams point out the exchangeability of residues in the proteins; the amino acids with similar diagrams have similar local folding requirements. PMID- 19280327 TI - Aged atria: electrical remodeling conducive to atrial fibrillation. AB - The incidence of atrial fibrillation (AF) increases with age. Alterations in structure and function of atrial ion channels associated with aging provide the substrate for AF. In this review we provide an overview of current knowledge regarding these age-related changes in atria, focusing on intrinsic ion channel function, impulse initiation and conduction. Studies on the action potentials (APs) of atria have shown that the AP contour is altered with age and the dispersion of AP parameters is increased with age. However, studies using human tissues are not completely consistent with experimental animal studies, since specimens from humans have been obtained from hearts with concomitant cardiovascular diseases and/or that are under the influence of pharmacologic agents. Ionic current studies show that while there are no age-related changes in sodium currents in atrial tissue, the calcium current is reduced and the transient outward and sustained potassium currents are increased in aged cells. While sinoatrial node firing is reduced with age, enhanced impulse initiation may occur in aged atrial cells, for example in the pulmonary veins and coronary sinus. Fibrous tissue is increased in aged atria, which is associated with an increased likelihood of abnormal electrical conduction. Thus, age-related AF involves alterations in the substrate as well as in the passive properties of aged atria. PMID- 19280328 TI - Complementary alternative medicine for children with autism: a physician survey. AB - Previous studies suggest over half of children with autism are using complementary alternative medicine (CAM). In this study, physicians responded (n = 539, 19% response rate) to a survey regarding CAM use in children with autism. Physicians encouraged multi-vitamins (49%), essential fatty acids (25%), melatonin (25%) and probiotics (19%) and discouraged withholding immunizations (76%), chelation (61%), anti-infectives (57%), delaying immunizations (55%) and secretin (43%). Physicians encouraging CAM were more likely to desire CAM training, inquire about CAM use, be female, be younger, and report greater autism visits, autism education and CAM knowledge. Physicians were more likely to desire CAM training, inquire about CAM and view CAM as a challenge for children with autism compared to children with other neurodevelopmental and chronic/complex conditions. PMID- 19280330 TI - Mal de Rio Cuarto virus (MRCV) genomic segment S3 codes for the major core capsid protein. AB - Mal de Rio Cuarto virus (MRCV) is a newly described species of the genus Fijivirus, family Reoviridae. Compared with other plant-infecting genus of the family, the function and localization of MRCV and other Fijivirus proteins are poorly understood. Through analysis of viral particle purifications, we positively identified five structural proteins of approximately 170, 140, 130, 66, and 62 kDa. The protein encoded by MRCV S3 genomic segment was expressed as a fusion protein in Escherichia coli, purified and used for rabbit immunization. The resulting antiserum reacted with the 140 kDa structural protein and with incomplete virus particles in immunoelectron microscopy assays, suggesting that MRCV S3 codes for the major core capsid protein. PMID- 19280331 TI - The association between body esteem and sexual desire among college women. AB - Relationships between body image variables and sexuality have been found among several groups of women. However, research has largely focused on generalized experiences of sexuality. With the exception of two studies which focused on specific medical populations, to our knowledge there has been no investigation of the relationship between body image and acute measures of sexual response. In the current study, we investigated the relationships between body esteem, sexual response to erotica in a laboratory-setting, and self-reported sexual functioning in a non-clinical sample of 85 college women. Women participated in one study session, during which mental sexual arousal, perceptions of physical arousal, and sexual desire were assessed. Results showed that higher body esteem was significantly positively related to sexual desire in response to erotica in the laboratory setting. Similarly, higher body esteem was positively related to self reported measures of sexual desire, as assessed by a validated measure of sexual function. The sexual attractiveness and weight concern subscales of the Body Esteem Scale, which relate to body characteristics that are most likely to be under public scrutiny, were particularly linked to sexual desire. This is the first study to show that body esteem is related to sexual responses to a standardized erotic stimulus in a laboratory setting. PMID- 19280333 TI - Access to primary care for homeless veterans with serious mental illness or substance abuse: a follow-up evaluation of co-located primary care and homeless social services. AB - To examine the hypothesis that a demonstration clinic integrating homeless, primary care, and mental health services for homeless veterans with serious mental illness or substance abuse would improve medical health care access and physical health status. A quasi-experimental design comparing a 'usual VA care' group before the demonstration clinic opened (N = 130) and the 'integrated care' group (N = 130). Regression models indicated that the integrated care group was more rapidly enrolled in primary care, received more prevention services and primary care visits, and fewer emergency department visits, and was not different in inpatient utilization or in physical health status over 18 months. The demonstration clinic improved access to primary care services and reduced emergency services but did not improve perceived physical health status over 18 months. Further research is needed to determine generalizability and longer term effects. PMID- 19280332 TI - Interaction of intensity and order regarding painful events. AB - While stimulus intensity obviously affects degree of pain responding, presentation order effects of stimuli of different intensities on acute pain responses are under-researched. The present study examined the effects of manipulating presentation order of lower and higher pain stimulus intensity. Using 96 undergraduates, this investigation employed a 2 x 2 mixed research design, with pain stimulus sequence as a between-subjects variable and pain stimulus trial as a repeated measure. When the greater pain stimulus intensity was presented last, verbal report of pain was higher. Also, performance of a cognitive task was interrupted the least when the lower stimulus intensity was presented last. Heart rate, however, was highest when the greater stimulus intensity was presented first, and pain tolerance was greatest when the lower stimulus intensity was presented first. Results are discussed in relation to adaptation-level effects, and implications for pain experienced in clinical settings are suggested. PMID- 19280334 TI - The role of harsh discipline in explaining sex differences in conduct disorder: a study of opposite-sex twin pairs. AB - In the current study, two hypotheses about the role of harsh discipline (HD) in explaining the sex difference in the prevalence of conduct disorder (CD) were evaluated: that boys exhibit more CD than girls because (1) they are exposed to more HD and/or (2) there is a greater association between HD and CD in boys. These hypotheses were evaluated in a sample of male and female adult twins from different families (N = 3,502) as well as a sample of adult twin brothers and sisters (N = 655) in order to examine the extent to which sex differences remained after controlling for between-family differences. Retrospective reports of HD experienced between ages 6-13 and DSM-IV CD symptoms experienced before age 18 were obtained via structured psychiatric telephone interviews. Boys reported higher mean levels of HD and CD than girls, both between and within families, and the results of model-fitting analyses suggested that differences in the use of harsh disciplinary practices for sons versus daughters may partially explain the sex difference in the prevalence of CD. Between families, the relation between HD and CD was greater for girls than boys, but within families, there was no evidence of a sex difference in the relation between HD and CD. Inconsistent between-family and within-family results suggest that factors that differ between families are confounded with sex differences in the relation between HD and CD. A more stringent test of sex differences involves eliminating these between-family differences by studying boys and girls within the same family. PMID- 19280335 TI - Awake EEG disregulation in good compared to poor sleepers. AB - This study was designed to test a disregulation model of sleep deprivation by assessing the ability of good sleepers compared to poor sleepers to shift daytime EEG patterning to changing environmental demands. Ten good and ten poor sleepers were identified from a sample of 110 college students who completed the Pittsburgh Sleep Quality Inventory (PSQI). EEG and SCR were recorded during a five task assessment session, including: (1) pre-baseline, (2) eyes open at rest, (3) eyes closed at rest, (4) sensory attentiveness (listening to an audio book clip), and (5) cognitive effort (a higher level cognitive flexibility task). A significant Group x Task interaction, F (3, 16) = 4.81, p = . 01 was attained on the theta data. Specifically, for good sleepers, theta decreased from the "eyes open at rest" to the "sensory attentiveness" tasks, while poor sleepers showed the opposite pattern. This pattern of theta suppression was found in 70% of the good sleepers and only 20% of the poor sleepers. No between group differences were noted in the SCR data, supporting a brain disregulation model, rather than a general psychophysiological stress model. PMID- 19280336 TI - Longitudinal study of nightmares in children: stability and effect of emotional symptoms. AB - Nightmares are defined as dreams with strong negative emotions which awaken the dreamer and are common during childhood: cross-sectional data shows the highest prevalence rates between the ages of five and ten. The present longitudinal study was designed to study the stability of nightmares over the course of 2 years. Sleep questionnaires and Strengths and Difficulties Questionnaires were completed by 851 10-years-old children and their parents, separately. In the total sample, nightmares occurred often in 2.5% (parental estimates) to 3.5% (self estimates of the children). The findings indicate that nightmare stability is considerably high, i.e., nightmare occurrence was predicted by the prevalence the year before (in addition to the concurrent amount of emotional symptoms). Children with 'chronic' nightmares showed more psychopathological symptoms. It would be interesting to follow-up these children in order to investigate whether childhood nightmares are also predictive for adult psychopathology. PMID- 19280337 TI - Assessment of parental experiential avoidance in a clinical sample of children with anxiety disorders. AB - This investigation seeks to establish the psychometric properties of an adapted measure of experiential avoidance (EA) in the parenting context by assessing its relation to other parenting constructs and psychosocial correlates of child anxiety in a clinical sample. Participants were 154 children (90 female, 64 male) diagnosed with anxiety disorders and their parents (148 mothers, 119 fathers). The newly developed Parental Acceptance and Action Questionnaire (PAAQ) was administered to parents along with self-report measures of adult experiential avoidance, parental psychopathology, affective expression, and parental control behaviors. A subsample of participants, n = 35, were re-administered the PAAQ to assess temporal stability. Factor analysis of the PAAQ yielded a two-factor solution with factors labeled Inaction and Unwillingness. Temporal stability of the PAAQ was found to be moderate, r = .68-.74. Internal consistency was fair across subscales of the PAAQ, alpha = .64-.65. Correlational analysis of the PAAQ and parent-report measures support the criterion validity of the PAAQ, suggesting that the PAAQ correlates with parent-report measures of parental locus of control, affective expression, and controlling parental behaviors as well as child psychopathology symptoms. Finally, the clinical applicability of the PAAQ is indicated by the PAAQ's ability to predict a significant amount of variance in parent- and clinician-rated levels of child anxiety and related psychopathology. PMID- 19280338 TI - Early-onset psychoses: comparison of clinical features and adult outcome in 3 diagnostic groups. AB - A comparison of clinical features and adult outcome in adolescents with three types of psychotic disorders: schizophrenic (SPh), schizoaffective (SA) and bipolar with psychotic features (BPP). Subjects (n = 41) were finally diagnosed (DSM-IV criteria) with SPh (n = 17), SA (n = 11) or BPP (n = 13). Clinical evaluation took place at onset and at a 3-year follow-up in all 41, and at least after 5 years in 36 patients. Symptoms were rated on the basis of the Positive and Negative Syndrome Scale (PANSS), integrating items from the Brief Psychiatric Rating Scale (BPRS) and the Kiddie Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime Version (K-SADS-PL). The Children Global Assessment Scale (C-GAS) and the Global Assessment Scale (GAF) were used to evaluate global functioning. Significant differences in clinical features were found in the three diagnostic groups as regards several parameters, some present on one and not on other rating scales, underscoring the insufficiency of a single scale for accurate analysis of the features of a psychotic disorder. At onset, a comparison using the simple presence/absence of symptoms showed scant differences among groups, while differences emerged if symptom severity was included in the comparison. Functioning at 3- and 5-year follow-ups showed a significantly better outcome in the BPP group and more substantial deterioration, with similar evolution, in the SPh and SA groups. The integration of several rating scales differentiated between diagnostic groups more effectively. The similar adult functioning outcome in the SPh and SA groups showed how difficult it is to clearly separate these two disorders. PMID- 19280339 TI - The diagnostic utility of behavioral checklists in identifying children with ADHD and children with working memory deficits. AB - The present study investigated whether children with ADHD and those with working memory impairments have a common behavioral profile in the classroom. Three teacher checklists were used: the Conners' teacher rating scale (CTRS), the behavior rating inventory of executive function (BRIEF), and the working memory rating scale. The Conners' continuous performance test (CPT) was also included to determine whether there is a correspondence between performance on this widely used cognitive measure of attention deficits and teacher ratings of classroom behavior. All three behavior scales, but not the CPT, were able to successfully discriminate children with ADHD and those with working memory deficits from typically-developing children. Both the CTRS and the BRIEF discriminated a significant proportion of the children with ADHD from those with working memory deficits, indicating that while both groups exhibit behavioral problems in the classroom, they are characterized by differential attention profiles. The children with ADHD were identified on the basis of oppositional and hyperactive behavior, while those with working memory deficits were more inattentive. PMID- 19280340 TI - Prognostic and predictive impact of central necrosis and fibrosis in early breast cancer: results from two International Breast Cancer Study Group randomized trials of chemoendocrine adjuvant therapy. AB - A minority of early invasive breast cancers show a pattern of central necrosis and fibrosis (CNF). Previous studies have documented an adverse prognostic impact and association with other adverse pathological features, but its predictive importance for therapy selection is unknown. We examined the prognostic and predictive value of CNF in two randomized clinical trials comparing chemoendocrine therapy with endocrine therapy alone in patients with node negative breast cancer. A total of 1,850 patients randomly assigned to treatment groups comparing endocrine with chemoendocrine therapy, and with centrally assessed CNF, ER, PgR and HER2 were included in the analytic cohort. The median follow up was 10 years. CNF was present in 84 of 1,850 trial patients (4.5%). It was associated with tumor characteristics suggesting poor outcome, but was an independent adverse factor for disease-free survival. In the presence of CNF outcome was worse regardless of tumor grade, whereas in the absence of CNF, patients with grade 3 tumors had poorer outcome than those with grade 1-2 tumors. Among patients with estrogen receptor-absent tumors chemoendocrine therapy was superior to endocrine therapy alone only in the absence of CNF [HR (chemoendocrine:endocrine) = 0.46 in CNF-absent, 0.90 in CNF-present], while among those with receptor-positive disease chemoendocrine therapy was beneficial only in the presence of CNF [HR = 0.34 CNF-present, 0.96 CNF-absent]. The results suggest that the presence of CNF reflects a biological difference in early breast cancer that is important in modulating the efficacy of standard therapies. Accordingly we believe that its presence should be routinely reported. PMID- 19280341 TI - Micro-electrocardiograms to study post-ventricular amputation of zebrafish heart. AB - The zebrafish (Danio rerio) is an emerging model for cardiovascular research. The zebrafish heart regenerates after 20% ventricular amputation. However, assessment of the physiological responses during heart regeneration has been hampered by the small size of the heart and the necessity of conducting experiments in an aqueous environment. We developed a methodology to monitor a real-time surface electrocardiogram (ECG) by the use of micro-electrodes, signal amplification, and a low pass-filter at a sampling rate of 1 kHz. Wavelet transform was used to further remove ambient noises. Rather than paralyzing the fish, we performed mild sedation by placing the fish in a water bath mixed with MS-222 (tricane methanesulfonate). We recorded distinct P waves for atrial contraction, QRS complexes for ventricular depolarization, and QT intervals for ventricular repolarization prior to, and 2 and 4 days post-amputation (dpa). Sedation reduced the mean fish heart rate from 149 +/- 18 to 90 +/- 17 beats/min. The PR and QRS intervals remained unchanged in response to ventricular apical amputation (n = 6, p > 0.05). Corrected QT intervals (QTc) were shortened 4 dpa (n = 6, p < 0.05). In a parallel study, histology revealed that apical thrombi were replaced with fibrin clots and collagen fibers. Atrial arrhythmia was noted in response to prolonged sedation. Unlike the human counterpart, ventricular tachycardia or fibrillation was not observed in response to ventricular amputation 2 and 4 dpa. Taken together, we demonstrated a minimally invasive methodology to monitor zebrafish heart function, electrical activities, and regeneration in real-time. PMID- 19280342 TI - Vessel diameter measurement from intravital microscopy. AB - The blood vessel diameter is often measured in microcirculation studies to quantify the effects of various stimuli. Intravital video microscopy is used to measure the change in vessel diameter by first recording the video and analyzing it using electronic calipers or by using image shearing technique. Manual measurement using electronic calipers or image shearing is time-consuming and prone to measurement error, and automated measurement can serve as an alternative that is faster and more reliable. In this paper, a new feature-based tracking algorithm is presented for automatically measuring diameter of vessels in intravital video microscopy image sequences. Our method tracks the vessel diameter throughout the entire image sequence once the diameter is marked in the first image. The parameters were calibrated using the intravital videos with manual ground truth measurements. The experiment with 10 synthetic videos and 20 intravital microscopy videos, including 10 fluorescence confocal and 10 non confocal transmission, shows that the measurement can be performed accurately. PMID- 19280343 TI - The role of posture, magnification, and grip force on microscopic accuracy. AB - While tremor has been studied extensively, the investigations thus far do not give detailed information on how the accuracy necessary for micromanipulations is affected while performing tasks in microsurgery and the life sciences. This paper systematically studies the effects of visual feedback, posture and grip force on the trial error and tremor intensity of subjects holding a forceps-like object to perform a pointing task. Results indicate that: (i) Arm support improves accuracy in tasks requiring fine manipulation and reduces tremor intensity in the 2-8 Hz region, but hand support does not provide the same effect; hence freedom of wrist movement can be retained without a significant increase in trial error. (ii) Magnification of up to x10 is critical to carry out accurate micromanipulations, but beyond that level, magnification is not the most important factor. (iii) While an appropriate grip force must be learned in order to grasp micro-objects, such as a needle, without damaging them, the level of grip force applied does not affect the endpoint accuracy. PMID- 19280344 TI - Mental health and addiction policy: what next? AB - The already frayed mental health and addictions safety net is in danger of collapse as demand for behavioral health services reaches an all-time high. The backdrop to our field's everyday struggles is health care reform. The details and implications of health care reform for the mental health and addictions community remain uncertain. What is certain is that we must build upon our 2008 legislative successes. While health care reform evolves, the National Council is focused on moving a practical actionable 2009 agenda that includes single points of accountability for continuity of care for people with serious mental illnesses and addictions, cost-based-plus financing that supports clinical excellence, a federal mental health funding stream dedicated to mental health and integrated treatment services for the uninsured, funds to support investments by behavioral health care organizations in information technology, and support for research based education and prevention practices. PMID- 19280345 TI - Permeation of four oral drugs through human intestinal mucosa. AB - The pharmaceutical industry is in need of rapid and accurate methods to screen new drug leads for intestinal permeability potential in the early stages of drug discovery. Excised human jejunal mucosa was used to investigate the permeability of the small intestine to four oral drugs, using a flow-through diffusion system. The four drugs were selected as representative model compounds of drug classes 1 and 3 according to the biopharmaceutics classification system (BCS). The drugs selected were zidovudine, propranolol HCl, didanosine, and enalapril maleate. Permeability values from our in vitro diffusion model were compared with the BCS permeability classification and in vivo and in vitro gastrointestinal drug permeability. The flux rates of the four drugs were influenced by the length of the experiment. Both class 1 drugs showed a significantly higher mean flux rate between 2 and 6 h across the jejunal mucosa compared to the class 3 drugs. The results are therefore in line with the drugs' BCS classification. The results of this study show that the permeability values of jejunal mucosa obtained with the flow-through diffusion system are good predictors of the selected BCS class 1 and 3 drugs' permeation, and it concurred with other in vitro and in vivo studies. PMID- 19280346 TI - Drug targeting to the hair follicles: a cyclodextrin-based drug delivery. PMID- 19280347 TI - Transbuccal delivery of 5-fluorouracil: permeation enhancement and pharmacokinetic study. AB - The purpose of this study was to determine the effect of permeation enhancers on the transbuccal delivery of 5-fluorouracil (FU). The effect of permeation enhancers on in vitro buccal permeability was assessed using sodium deoxycholate (SDC), sodium dodecyl sulphate (SDS), sodium tauroglycocholate (STGC), and oleic acid and their concentrations for absorption enhancement were optimized. STGC appeared to be most effective for enhancing the buccal permeation of FU than the other enhancers. These enhancements by STGC were statistically significant (p < 0.05) compared to control. The order of permeation enhancement was STGC > SDS > SDC > oleic acid. Histological investigations were performed on buccal mucosa and indicated no major morphological changes. The enhancing effect of STGC on the buccal absorption of FU was evaluated from the mucoadhesive gels in rabbits. The absolute bioavailability of FU from mucoadhesive gels containing STGC increased 1.6-fold as compared to the gels containing no permeation enhancer. The mean residence time and mean absorption time considerably increased following administration of gel containing penetration enhancer compared with the gel without penetration enhancer. PMID- 19280348 TI - The abbreviated impactor measurement (AIM) concept: part 1--Influence of particle bounce and re-entrainment-evaluation with a "dry" pressurized metered dose inhaler (pMDI)-based formulation. AB - The abbreviated impactor measurement concept is a potential improvement to the labor-intensive full-resolution cascade impactor methodology for inhaler aerosol aerodynamic particle size distribution (APSD) measurement by virtue of being simpler and therefore quicker to execute. At the same time, improved measurement precision should be possible by eliminating stages upon which little or no drug mass is collected. Although several designs of abbreviated impactor systems have been developed in recent years, experimental work is lacking to validate the technique with aerosols produced by currently available inhalers. In part 1 of this two-part article that focuses on aerosols produced by pressurized metered dose inhalers (pMDIs), the evaluation of two abbreviated impactor systems (Copley fast screening Andersen impactor and Trudell fast screening Andersen impactor), based on the full-resolution eight-stage Andersen nonviable cascade impactor (ACI) operating principle, is reported with a formulation producing dry particles. The purpose was to investigate the potential for non-ideal collection behavior associated with particle bounce in relation to internal losses to surfaces from which particles containing active pharmaceutical ingredient are not normally recovered. Both abbreviated impactors were found to be substantially equivalent to the full-resolution ACI in terms of extra-fine and fine particle and coarse mass fractions used as metrics to characterize the APSD of these pMDI produced aerosols when sampled at 28.3 L/min, provided that precautions are taken to coat collection plates to minimize bounce and entrainment. PMID- 19280349 TI - Quercetin/beta-cyclodextrin solid complexes prepared in aqueous solution followed by spray-drying or by physical mixture. AB - The present study was designed to investigate the influence of operating conditions (temperature, stirring time, and excess amount of quercetin) on the complexation of quercetin with beta-cyclodextrin using a 2(3) factorial design. The highest aqueous solubility of quercetin was reached under the conditions 37 degrees C/24 h/6 mM of quercetin. The stoichiometric ratio (1:1) and the apparent stability constant (Ks = 230 M(-1)) of the quercetin/beta-cyclodextrin complex were determined using phase-solubility diagrams. The semi-industrial production of a 1:1 quercetin/beta-cyclodextrin solid complex was carried out in aqueous solution followed by spray-drying. Although the yield of the spray-drying process was adequate (77%), the solid complex presented low concentration of quercetin (0.14%, w/w) and, thus, low complexation efficiency. The enhancement of aqueous solubility of quercetin using this method was limited to 4.6-fold in the presence of 15 mM of beta-cyclodextrin. Subsequently, an inclusion complex was prepared via physical mixture of quercetin with beta-cyclodextrin (molar ratio of 1:1 and quercetin concentration of 23% (w/w)) and characterized using infrared spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, and scanning electron microscopy analyses. The enhancement of aqueous solubility of quercetin using this method was 2.2-fold, similar to that found in the complex prepared in aqueous solution before the spray-drying process (2.5-fold at a molar ratio of 1:1, i.e., 6 mM of quercetin and 6 mM of beta cyclodextrin). PMID- 19280350 TI - Physicochemical characteristics, cytotoxicity, and antioxidant activity of three lipid nanoparticulate formulations of alpha-lipoic acid. AB - Exogenously supplied alpha-lipoic acid (LA) has proven to be effective as an antioxidant. In an effort to develop a water-soluble formulation for topical administration, LA was formulated in the form of solid lipid nanoparticles (SLN), nanostructure lipid carriers (NLC), and nanoemulsion (NE) and characterized in terms of physical and biological properties. Mean particle size of 113, 110, and 121 nm were obtained for NE, NLC, and SLN, respectively, with narrow size distribution. Zeta potential was approximately in the range of -25 to -40 mV. Disc and spherical structures of nanoparticles were observed by cryo-scanning electron microscopy. Entrapment efficiency of LA in three formulations was found to be more than 70%. After 120 days of storage at 25 degrees C, physical stability of all formulations remained unchanged whereas the entrapment efficiency of SLN and NLC could be maintained, suggesting relative long-term stability. Prolonged release of LA formulation following the Higuchi model was found where a faster release was observed from NE compared with that of SLN and NLC. More than 80% of cell survivals were found up to 1 microM of LA concentrations. Antioxidant activity analysis demonstrated that all LA-loaded formulations expressed antioxidant activity at a similar magnitude as pure LA. These results suggest that chosen compositions of lipid nanoparticles play an important role on drug loading, stability, and biological activity of nanoparticles. Both SLN and NLC demonstrated their potential as alternative carriers for aqueous topical administration of LA. PMID- 19280351 TI - Nicotine is a selective pharmacological chaperone of acetylcholine receptor number and stoichiometry. Implications for drug discovery. AB - The acronym SePhaChARNS, for "selective pharmacological chaperoning of acetylcholine receptor number and stoichiometry," is introduced. We hypothesize that SePhaChARNS underlies classical observations that chronic exposure to nicotine causes "upregulation" of nicotinic receptors (nAChRs). If the hypothesis is proven, (1) SePhaChARNS is the molecular mechanism of the first step in neuroadaptation to chronic nicotine; and (2) nicotine addiction is partially a disease of excessive chaperoning. The chaperone is a pharmacological one, nicotine; and the chaperoned molecules are alpha4beta2* nAChRs. SePhaChARNS may also underlie two inadvertent therapeutic effects of tobacco use: (1) the inverse correlation between tobacco use and Parkinson's disease; and (2) the suppression of seizures by nicotine in autosomal dominant nocturnal frontal lobe epilepsy. SePhaChARNS arises from the thermodynamics of pharmacological chaperoning: ligand binding, especially at subunit interfaces, stabilizes AChRs during assembly and maturation, and this stabilization is most pronounced for the highest-affinity subunit compositions, stoichiometries, and functional states of receptors. Several chemical and pharmacokinetic characteristics render exogenous nicotine a more potent pharmacological chaperone than endogenous acetylcholine. SePhaChARNS is modified by desensitized states of nAChRs, by acid trapping of nicotine in organelles, and by other aspects of proteostasis. SePhaChARNS is selective at the cellular, and possibly subcellular, levels because of variations in the detailed nAChR subunit composition, as well as in expression of auxiliary proteins such as lynx. One important implication of the SePhaChARNS hypothesis is that therapeutically relevant nicotinic receptor drugs could be discovered by studying events in intracellular compartments rather than exclusively at the surface membrane. PMID- 19280352 TI - Modelling and PBPK simulation in drug discovery. AB - Physiologically based pharmacokinetic (PBPK) models are composed of a series of differential equations and have been implemented in a number of commercial software packages. These models require species-specific and compound-specific input parameters and allow for the prediction of plasma and tissue concentration time profiles after intravenous and oral administration of compounds to animals and humans. PBPK models allow the early integration of a wide variety of preclinical data into a mechanistic quantitative framework. Use of PBPK models allows the experimenter to gain insights into the properties of a compound, helps to guide experimental efforts at the early stages of drug discovery, and enables the prediction of human plasma concentration time profiles with minimal (and in some cases no) animal data. In this review, the application and limitations of PBPK techniques in drug discovery are discussed. Specific reference is made to its utility (1) at the lead development stage for the prioritization of compounds for animal PK studies and (2) at the clinical candidate selection and "first in human" stages for the prediction of human PK. PMID- 19280353 TI - Molecular and clinical aspects of endothelial dysfunction in diabetes. AB - Diabetic patients have an increased risk for cardiovascular complications with respect to the general population. Micro- and macrovascular complications such as nephropathy, retinopathy, atherosclerosis, and coronary artery disease are usually preceded by endothelial dysfunction, a condition characterized by impaired vasorelaxation resulting from reduced bioavailability of the endothelial mediator nitric oxide (NO). Nitric oxide is among endothelial mediators released by endothelial cells in response to insulin stimulation. Therefore, metabolic abnormalities such as insulin resistance, dyslipidemia, compensatory hyperinsulinemia and overt hyperglycemia may all contribute to impaired NO bioavailability and abnormal vasodilatation in diabetic patients. Each of these alterations may trigger endothelial dysfunction by multiple intracellular mechanisms including accelerated formation of advanced glycolysis end products, activation of protein kinase C, increased pro-inflammatory signaling, and impaired sensitivity of the PI 3-kinase signaling pathways. This review outlines the most important mechanisms by which insulin takes part in physiological regulation of endothelial function. Abnormal insulin signaling in endothelium under diabetic conditions and patho-physiological consequences on cardiovascular homeostasis will also be discussed. PMID- 19280354 TI - Nutritionally relevant elements in staple foods: influence of arable site versus choice of variety. AB - Eighteen representative sites for the Austrian grain-growing and eight for the potato-growing zones (soils and crops) were investigated. On each site, total element contents (B, Ba, Ca, Cd, Co, Cu, Fe, K, Li, Mg, Mn, Mo, Na, P, Sr and Zn) were determined in 4-12 varieties of winter wheat (n = 136), 6 varieties of spring durum wheat (n = 30), 5 varieties of winter durum wheat (n = 15), 7 varieties of rye (n = 49), 5 varieties of spring barley (n = 30) and 5 varieties of potatoes (n = 40). Element accumulations in grain species and potato tubers varied significantly with site conditions, with the main exceptions for B in potatoes and wheat as well as for Zn, Cu and Co in durum wheat. On average, across all investigated sites, differences in varieties occurred concerning the elements Ca, Cd, Ba, Sr and Zn (except Zn in potatoes and winter durum). A rough estimation revealed that an average Austrian consumer of wheat, rye and potatoes meets more than 50% of the needs of daily element intake for K, P and Mg, between 36 and 72% for Fe, Zn and Cu, and more than 100% for Co, Mo and Mn. In particular, the elements Ca and Na have to be added from other sources. PMID- 19280355 TI - Proportionate cancer incidence in the Laotian population of California, 1988 2006. AB - When the Vietnam War ended in 1975, pro-US Laotians (including Lao, Mien, Khmu) were displaced and became refugees in their own native country. Thousands fled to refugee camps in nearby Thailand and were eventually relocated to several Western countries, including the US. A listing of 1,195 Laotian cancer cases were extracted from the California Cancer Registry for diagnosis years 1988-2006. Cancer cases with birthplace coded as "Laos" were included. Proportionate incidence ratios (PIRs) and associated 95% confidence intervals (CI) were calculated for 17 selected cancer sites. The total population of California (all race/ethnic groups combined) was used as the reference. Proportional occurrence of cancers varied by genders and by cancer sites. Laotians in California experienced statistically significantly elevated risks for cancer of the nasopharynx (PIR = 14.8; 95% CI = 10.5-20.1), liver (PIR = 12.6; 95% CI = 10.8 14.6), stomach (PIR = 3.1; 95% CI = 2.4-4.0), cervix (PIR = 1.9; 95% CI = 1.5 2.3), pancreas (PIR = 2.1; 95% CI = 1.5-2.8), oral cavity (PIR = 1.8; 95% CI = 1.4-2.3), lung and bronchus (PIR = 1.4; 95% CI = 1.2-1.7). As found for other Asian subgroups, Laotians, too, have statistically significantly reduced risks for colorectal (PIR = 0.8; 95% CI = 0.6-0.9), colon (PIR = 0.7; 95% CI 0.5-0.9), breast (PIR 0.7; 95% CI = 0.5-0.8), and prostate (PIR = 0.1; 95% CI = 0.0-0.2) cancers. The increased risk found for mostly non-Western types of cancers have implications for culturally responsive cancer control and intervention activities targeting the Laotian population. PMID- 19280356 TI - Associations between endogenous sex hormone levels and mammographic and bone densities in premenopausal women. AB - PURPOSE: Mammographic breast and bone mineral densities (BMD) have been associated with luteal phase hormone concentrations in premenopausal women. We assessed the associations of breast and bone densities with follicular phase hormones and sex hormone binding globulin (SHBG) in premenopausal women, given that follicular phase hormones have been shown to be positively associated with premenopausal breast cancer risk. METHODS: One hundred and ninety-two 40-45-year old women provided a spot urine and/or blood sample during the follicular phase. Hormone and SHBG concentrations, and bone density were measured and routine mammograms were accessed and digitized to obtain breast density measures. Regression models were fit to assess the associations between hormones and SHBG, and breast and bone densities. RESULTS: Positive associations were observed between percent breast density and SHBG (p trend = 0.02), as well as estradiol (p trend = 0.08), after controlling for body mass index (BMI), number of pregnancies, and breast feeding history. In addition, a statistically significant inverse association was observed between total testosterone and head BMD (p trend = 0.01), after controlling for BMI. CONCLUSIONS: Associations were observed between breast and bone densities, and serum hormone concentrations during the follicular phase of the menstrual cycle. PMID- 19280357 TI - Application of CMB model for source apportionment of polycyclic aromatic hydrocarbons (PAHs) in coastal surface sediments from Rizhao offshore area, China. AB - Polycyclic aromatic hydrocarbons (PAHs) in coastal surface sediments from Rizhao offshore area were analyzed by gas chromatography-mass spectrometry. A chemical mass balance (CMB) model developed by the U.S. Environmental Protection Agency (EPA), CMB8.2, was used to apportion sources of PAHs. Seven possible sources, including coal residential, coal power plant, diesel engines exhaust, gasoline engines exhaust, coke oven, diesel oil leaks, and wood burning, were chosen as the major contributors for PAHs in coastal surface sediments. To establish the fingerprints of the seven sources, source profiles were collected from literatures. After including degradation factors, the modified model results indicate that diesel oil leaks, diesel engines exhaust, and coal burning were the three major sources of PAHs. The source contributions estimated by the EPA's CMB8.2 model were 9.25%, 15.05%, and 75.70% for diesel oil leaks, diesel engines exhaust, and coal burning, respectively. PMID- 19280358 TI - Spatiotemporal analysis of the PM10 concentration over the Taranto area. AB - In this paper, an analysis of air quality data is provided for the municipal area of Taranto (southern Italy) characterized by high environmental risks as formally decreed by the Italian government in the 1990s with two administrative measures. This is due to the massive presence of industrial sites with elevated environmental impact activities along the NW boundary of the city conurbation. The aforementioned activities have effects on the environment and on public health, as a number of epidemiological researches concerning this area reconfirm. The present study is focused on particulate matter as measured by PM10 concentrations at 13 monitoring stations, equipped with analogous instruments based on the Beta absorption technology, either reporting hourly, two-hourly, or daily measurements. Daily estimates of the PM10 concentration surfaces are obtained in order to identify areas of higher concentration (hot spots), possibly related to specific anthropic activities. Preliminary analysis involved addressing several data problems: (1) due to the use of two different validation techniques, a calibration procedure was devised to allow for data comparability; (2) imputation techniques were considered to cope with the large number of missing data, due to both different working periods and occasional malfunctions of PM10 sensors; and (3) reliable weather covariates (wind speed and direction, pressure, temperature, etc.) were obtained and considered within the analysis. Spatiotemporal modelling was addressed by a Bayesian kriging-based model proposed by Le and Zidek (2006) characterized by the use of time varying covariates and a semiparametric covariance structure. Advantages and disadvantages of the model are highlighted and assessed in terms of fit and performance. Estimated daily PM10 concentration surfaces are suitable for the interpretation of time trends and for identifying concentration peaks within the urban area. PMID- 19280359 TI - Semen biology of vendace (Coregonus albula L.). AB - The objective of this study was to describe the morphometry and motility parameters of vendace (Coregonus albula) spermatozoa. Morphometric parameters of vendace sperm head and tail were of values similar to rainbow trout. The effects of pH, sodium, potassium and calcium ion concentrations on computer-assisted sperm analysis (CASA) sperm motility characteristics were tested. Vendace sperm was motile in a wide pH range of 6.0-10.5 with the optimum pH established at 9.0. Increases in potassium and calcium ions caused decreases in the percentage of motile sperm. The CASA parameters and erratic sperm movement pattern of vendace spermatozoa were similar to whitefish (C. lavaretus) sperm motility, suggesting that there is a coregonid-specific sperm motility pattern. PMID- 19280360 TI - Impact of acute kidney injury on metformin-associated lactic acidosis. AB - OBJECTIVES: Metformin has been shown to reduce diabetic complications in overweight patients, and is increasingly used to treat this condition. However, this agent is associated with a rare but serious risk of lactic acidosis. METHODS: From January 2004 to November 2008, 11 cases of metformin-associated lactic acidosis were seen at a medical center in Taiwan. The patients' records were retrospectively reviewed with respect to clinical presentation, biochemical data, therapeutic strategies, and outcomes. RESULTS: Ten out of 11 patients with acute kidney injury were enrolled in this study. From their pre-existing states, these patients would have been considered at low risk of developing lactic acidosis. However, all of them had acute intercurrent conditions that might compromise renal function. Among them, six patients were ventilated and five patients had severe hypotension with vasoactive support. Nine patients received renal replacement therapy to clear the acidosis and treat the renal failure. All but one of our patients survived. Recovery of renal function was the general rule in survivors. CONCLUSIONS: Metformin-associated lactic acidosis is commonly accompanied with acute kidney injury. In spite of the severity of their illness, the outcome is favorable with intensive support of the cardiovascular, respiratory, and renal systems. PMID- 19280361 TI - The comparison of Belt-Fuqua and "TIPU" techniques in proximal penile hypospadias cases: retrospective analysis of 65 cases. AB - OBJECTIVE: We have compared our success and complication rates using the single step "TIPU" (tubularized incised plate urethroplasty) and two-step Belt-Fuqua techniques in patients with proximal penile hypospadias. PATIENTS AND METHODS: Sixty-five patients with proximal penile hypospadias were operated on in our clinic between 1998 and 2008: 31 by the "TIPU" technique and 34 by the Belt-Fuqua technique. The correction of the chordee was accomplished with dorsal plication and corporal body grafting in 18 patients in the "TIPU" group and at the first stage in all patients in the Belt-Fuqua group. RESULTS: The mean age of the patients was 6.2 years in the "TIPU" group and 6.6 years in the Belt-Fuqua group. The mean follow-up duration was 9.2 and 7.9 months, respectively. The overall complication rate was 22.6% in the "TIPU" group and 11.8% in the Belt-Fuqua group. In both groups, the cosmetic appearance of a natural vertical slit glanular meatus situated at the normal position on the glans was achieved. CONCLUSION: The two-step Belt-Fuqua technique is accepted as a good alternative treatment modality because of low complication rates and good results in selected cases with proximal penile hypospadias. PMID- 19280362 TI - Cryptic species in the Terfezia boudieri complex. AB - Phylogenetic analyses have corroborated the discovery of three internal transcribed spacer (ITS) Types in Terfezia boudieri isolates in the course of earlier studies and have emphasized the divergence of Type 2 from Types 1 and 3. The application of molecular and physiological tools described below, revealed the existence of cryptic species within T. boudieri. The markers used include sequences taken from the 5' end of the ribosomal large subunit gene, a chitin synthase partial sequence, beta-tubulin partial sequence and amplified fragment length polymorphism (AFLP)-based markers. Following initial sequencing of a single PCR amplified sample for each Type, mass analysis of specimens relied on RFLP differences between the Types. Over 100 fruit bodies, 30 or more specimens for each ITS Type, were tested with each of the markers. The markers analysis divided the isolates into three groups, each correlated to a specific ITS Type. Two of the physiological traits examined: mycelial proliferation and mycorrhiza formation, consistently showed responses paralleling the ITS Types; the data presented suggest that T. boudieri is comprised of three cryptic species. PMID- 19280363 TI - Reliability of the medication appropriateness index in Dutch residential home. AB - OBJECTIVE: To validate the reliability of the Medication Appropriateness Index in the appraisal of quality of prescribing by assessing the inter- and intra-group variation in Dutch residential home patients. SETTING AND METHOD: Eight raters evaluated 81 medication records of 15 patients from a group of older patients living in a residential nursing home The Netherlands. These patients had been recruited for a medication review investigation throughout a period of 12 months over the period April 2003 until April 2004. Patient information was acquired by connecting the medical record with the complete prescription record and pharmaceutical record. Each patient was assessed twice by two independent reviewers on the basis of a patient profile in combination with the extracted medical record and using a structured procedure. MAIN OUTCOME: A summed MAI score, percent agreement, kappa, positive and negative agreement as well as intra class correlation coefficient were calculated for each criterion. MEASURE: Medication appropriateness was assessed with the Medication Appropriateness Index (MAI) by an independent panel of Dutch hospital and community pharmacists. RESULTS: The overall percentage agreement was 83%. For each of the 10 different medication appropriateness questions it ranged from 79 to 100% for appropriate and from 47 to 60% for inappropriate ratings. The overall chance adjusted inter rater agreement reached a moderate kappa score of 0.47. The overall intra-group agreement was very good with an overall percentage of 98 and a kappa score of at least 0.84 (all schemes). CONCLUSION: In a Dutch institutionalised setting with representative raters, the unmodified MAI can be used as an instrument to quantify changes in appropriateness of prescribing. PMID- 19280364 TI - Four successful pregnancies in a patient with mucopolysaccharidosis type I treated by allogeneic bone marrow transplantation. AB - To date, little is known about the fertility of women suffering from mucopolysaccharidosis type I (MPS I). We report on a female patient with MPS I treated by allogeneic bone marrow transplantation (BMT) at the age of 4 years (after a conditioning regimen containing busulfan 16 mg/kg and cyclophosphamide 100 mg/kg) who had four successful pregnancies without any reproductive assistance. Clinical and biological examinations of the children were normal. On the basis of this case, we discuss the fertility counselling of female MPS I patients at the time of BMT. PMID- 19280366 TI - The differential expression of peroxisome proliferators-activated receptors in various duck tissues. AB - Peroxisome proliferators-activated receptors (PPARs) plays important role in regulate a variety of target genes which involves in lipid metabolism. In the present study, 1,430 bp of PPARalpha and 1,650 bp of PPARgamma in Peking duck (Anas platyrhynchos) were cloned by reverse transcription (RT)-PCR. The cloned sequences of PPARalpha and PPARgamma were highly conserved, although relevant changes with respect to human, mouse, chicken, and goose were identified. Real time quantitative RT-PCR was used to determine if the PPARalpha and PPARgamma were expressed in different tissues of male and female Peking ducks. The expression of PPARalpha and PPARgamma were widely distributed in all examined tissues. The expression level of PPARalpha was highest in liver and lowest in kidney in male and female ducks. And it was highest in abdominal fat and lowest in small intestine of PPARgamma in male and female ducks. PMID- 19280367 TI - Different properties of P2X(7) receptor in hippocampal and cortical astrocytes. AB - P2X(7) receptor is a ligand-gated ion channel, which can induce the opening of large membrane pores. Here, we provide evidence that the receptor induces pore formation in astrocytes cultured from cortex, but not from the hippocampus. Furthermore, P2X(7) receptor activation promptly induces p38 mitogen-activated protein kinase (MAPK) phosphorylation in cortical but not in hippocampal astrocytes. Given the role of p38 MAPK activation in pore opening, these data suggest that defective coupling of the receptor to the enzyme could occur in hippocampal cultures. The different capabilities of the receptor to open membrane pores cause relevant functional consequences. Upon pore formation, caspase-1 is activated and pro-IL1-beta is cleaved and released extracellularly. The receptor stimulation does not result in interleukin-1beta secretion from hippocampal astrocytes, although the pro-cytokine is present in the cytosol of lipopolysaccharide-primed cultures. These results open the possibility that activation of P2X(7) receptors differently influences the neuroinflammatory processes in distinct brain regions. PMID- 19280368 TI - Protein synthesis rates in Drosophila associate with levels of the hsr-omega nuclear transcript. AB - Transcripts of the Drosophila hsr-omega gene are known to interact with RNA processing factors and ribosomes and are postulated to aid in co-ordinating nuclear and cytoplasmic activities particularly in stressed cells. However, the significance of these interactions for physiological processes and in turn for whole-organism fitness remains an open question. Because hsr-omega's cellular expression characteristics suggest it may influence protein synthesis, and because both genotypic and expression variation of hsr-omega have been associated with thermotolerance, we characterised 30 lines for variation in the rates of protein synthesis, measured in ovarian tissues, both before and after a mild heat shock, and for basal levels of the two main hsr-omega transcripts, omega-n and omega-c. As expected, the mild heat shock reduced protein synthesis rates. Large variation occurred among lines in levels of omega-n which was negatively associated with rates of basal protein synthesis--a result that supports the model for the cellular function of omega-n. Furthermore, omega-n levels were associated with hsr-omega genotype of the line parents. Little variation occurred among lines for omega-c levels and no associations were detected with protein synthesis or genotype. Since protein synthesis is a fundamental process for growth and development, we characterised the lines for several life-history traits; however, no associations with protein synthesis, omega-n or omega-c levels were detected. Our results are consistent with the idea that natural variation in hsr-omega expression influence rates of protein synthesis in this species. PMID- 19280370 TI - Pulmonary function testing in the evaluation of patients with allergic and immunologic disorders. PMID- 19280369 TI - Cloning, characterization, and functional studies of a human 40-kDa catecholamine regulated protein: implications in central nervous system disorders. AB - Catecholamine-regulated proteins (CRPs) have been shown to bind dopamine and other structurally related catecholamines; in particular, the 40-kDa CRP (CRP40) protein has been previously cloned and functionally characterized. To determine putative human homologs, BLAST analysis using the bovine CRP40 sequence identified a human established sequence tag (EST) with significant homology (accession #BQ224193). Using this EST, we cloned a recombinant human brain CRP40 like protein, which possessed chaperone activity. Radiolabeled dopamine binding studies with recombinant human CRP40 protein demonstrated the ability of this protein to bind dopamine with low affinity and high capacity. The full-length human CRP40 nucleotide sequence was elucidated (accession #DQ480334) with RNA ligase-mediated rapid amplification of complementary DNA ends polymerase chain reaction, while Northern blot hybridization suggested that human CRP40 is an alternative splice variant of the 70-kDa mitochondrial heat shock protein, mortalin. Human SH-SY5Y neuroblastoma cells treated with the antipsychotic drug, haloperidol, exhibited a significant increase in CRP40 messenger RNA expression compared to untreated control cells, while other dopamine agonists/antagonists also altered CRP40 expression and immunolocalization. In conclusion, these results show that we have cloned a splice variant of mortalin with a novel catecholamine binding function and that this chaperone-like protein may be neuroprotective in dopamine-related central nervous system disorders. PMID- 19280371 TI - Gonadotropin-independent precocious puberty associated with a somatic activating mutation of the LH receptor gene: detection of a mutation present in only a small fraction of cells from testicular tissue using wild-type blocking polymerase chain reaction and laser-capture microdissection. AB - OBJECTIVE: Leydig cells are the principal source of testosterone, and boys with Leydig cell tumors typically have signs of gonadotropin-independent precocious puberty as a result of testosterone secretion by the tumor. A single somatic activating mutation of the LH receptor gene, Asp578His, limited to the tumoral Leydig cells, has been described in a few boys with gonadotropin-independent precocious puberty. We report a molecular study of a boy with gonadotropin independent precocious puberty caused by a Leydig cell tumor. DESIGN AND SETTING: This is a clinical case report from the Kobe Children's Hospital. PATIENT AND METHODS: One patient with gonadotropin-independent precocious puberty caused by a Leydig cell tumor underwent a left orchidectomy. We performed a genetic study of the tumoral Leydig cells. RESULT: Using wild-type blocking PCR (WTB-PCR) and laser-capture microdissection (LCM), we found that the Asp578His mutation of the LH receptor gene was exclusively localized to the tumoral Leydig cells and was absent in the adjacent normal tissue and leukocytes. CONCLUSIONS: WTB-PCR and LCM are powerful techniques that can detect a somatic mutation present in only a small fraction of cells from heterozygous tissue samples. PMID- 19280372 TI - Anticancer effect of realgar nanoparticles on mouse melanoma skin cancer in vivo via transdermal drug delivery. AB - Realgar has been used successfully to treat diseases for thousands of years, but its poor water solubility and high toxicity hampered its further medical uses. Here, we first applied transdermal drug delivery system to deliver realgar nanoparticles to investigate its anticancer effect and toxicity in vivo. In this study, MTT assay and flow cytometry analysis demonstrated that realgar significantly suppressed the proliferation and induced apoptosis of B16 melanoma cells in a dose-dependent manner. Transdermal penetration studies in vitro showed realgar nanoparticles could be delivered efficiently through skin. Tests on tumor bearing C57BL/6 mice displayed that realgar could decrease the tumor volume markedly via transdermal drug delivery compared with the intraperitoneal administration and the control. Hematoxylin-eosin and immunohistochemical staining revealed that it could inhibit angiogenesis. The monitoring of the hepatic injury, body weight, feeding behavior, motor activity, and skin irritation of each animal indicated little toxicity of realgar to mice. The results demonstrated that realgar nanoparticles can be dermally delivered to achieve high efficacy against menaloma in vivo with low toxicity. PMID- 19280373 TI - Mitral annulus caseous calcification imaged with 64-slice MDCT. PMID- 19280374 TI - The plasma zinc/serum copper ratio as a biomarker in children with autism spectrum disorders. AB - The frequency of zinc deficiency, copper toxicity and low zinc/copper in children with autism spectrum disorders (ASDs) may indicate decrement in metallothionein system functioning. A retrospective review of plasma zinc, serum copper and zinc/copper was performed on data from 230 children with autistic disorder, pervasive developmental disorder-NOS and Asperger's syndrome. The entire cohort's mean zinc level was 77.2 microg dl(-1), mean copper level was 131.5 microg dl( 1), and mean Zn/Cu was 0.608, which was below the 0.7 cut-off of the lowest 2.5% of healthy children. The plasma zinc/serum copper ratio may be a biomarker of heavy metal, particularly mercury, toxicity in children with ASDs. PMID- 19280375 TI - The motor profile of primary school-age children with a 22q11.2 deletion syndrome (22q11.2DS) and an age- and IQ-matched control group. AB - In the early publications on the 22q11.2 Deletion Syndrome (22q11.2DS) motor abnormalities have been frequently reported. However, systematic studies on the motor performance of children with the 22q11.2DS, and especially of school-age children, are scarce. In this study the motor performance of primary school-age children with a 22q11.2DS (n = 28) was compared with an age- and IQ-matched control group (n = 28) using the Movement Assessment Battery for Children (MABC), the Korperkoordinationstest fur Kinder (KTK) and the Beery-Buctenica test of Visual-Motor Integration (Beery). Children with a 22q11.2DS scored significantly lower than the age- and IQ-matched control group on the subsection Manual Dexterity (MABC) and the Visual Perception and Motor Coordination subtests of the Beery. When investigating the correlations between Intelligence quotient (IQ) and motor performance, a specific profile was found in the 22q11.2DS group when compared with the age- and IQ-matched control group. Because an IQ-matched control group was adopted, the deficits in visual-perceptual and visuomotor integration skills cannot fully be attributed to a general developmental delay and thus may be specific for the 22q11.2DS. Future studies that investigate the specificity of the visual-perceptual problems - both on the behavioral and brain level (functional Magnetic Resonance Imaging [fMRI] and Diffusion Tensor Imaging [DTI]) - are necessary to answer this question. Nonetheless, the importance of incorporating motor functioning into the study of the neuropsychological profile of children with a 22q11.2DS has to be stressed. PMID- 19280376 TI - Effect of chemical enhancers on percutaneous absorption of daphnetin in isopropyl myristate vehicle across rat skin in vitro. AB - The percutaneous absorption properties of daphnetin with chemical penetration enhancers were investigated to explore the feasibility of daphnetin as a candidate for transdermal delivery to treat arthritis. Permeation experiments were carried out in vitro using 2-chamber diffusion cells in isopropyl myristate (IPM) vehicle using rat abdominal skin as a barrier. Various enhancers were employed, including O-acylmenthol derivatives synthesized in the laboratory and many conventional enhancers. Among the O-acylmenthol derivatives, 2-isopropyl-5 methylcyclohexyl 2-hydroxypanoate (M-LA) demonstrated a significant enhancing effect on daphnetin permeation. The highest degree of enhancement was obtained when NMP combined with Span 80 and the cumulative transport was 667.29 microg/cm(2) over 8 h. The solubility parameters, vehicle/stratum corneum partition, and diffusion coefficients were calculated to clarify the enhancing mechanism of classic enhancers on daphnetin. In conclusion, these findings allow a rational approach for designing an effective daphnetin transdermal delivery system. PMID- 19280378 TI - A two-week stay in an Interprofessional Training Unit changes students' attitudes to health professionals. AB - Attitudes amongst health professionals can impact on the effectiveness of teamworking and patient care. Interprofessional education (IPE) is thought to contribute to the development of positive attitudes. An Interprofessional Training Unit (ITU) was set up to create an optimal learning environment for healthcare students. Students' attitudes were assessed, using a version of the Attitudes to Health Professionals Questionnaire (AHPQ) that had been translated into the students' native language. This paper describes the process undertaken to obtain a trustworthy translation. One hundred and sixty two students from occupational therapy, physiotherapy, medicine and nursing completed the questionnaire before and after their stay at the ITU. Their responses were analysed in relation to the constructs "caring" and "subservient" from the validated English version of AHPQ. Echoing earlier studies elsewhere, at the beginning students viewed doctors as being the least "caring" and the least "subservient" professional group. Nurses were seen by the students as being the most "caring" and the most "subservient" profession. After the ITU experience students viewed most professions as more "caring" and less "subservient", apart from doctors, who were seen as being more subservient after the ITU experience. This study indicates that an IPE initiative such as the ITU can impact positively on students' attitudes and that the translated AHPQ can be used to monitor this attitudinal change. PMID- 19280379 TI - Non-right-handedness and mental health problems among adolescents from the general population: The Trails Study. AB - To determine whether the association between non-right-handedness and mental problems among adolescents is specific for psychotic symptoms, we included a group of 2096 adolescents with a mean age of 14 years from the general population. Mental health problems were assessed using the parent, self-report, and teacher versions of the Child Behavior Checklist. Internalising problems comprised anxious and depressed, withdrawn and depressed, and somatic complaints. Externalising problems consisted of delinquent behaviour and aggressive behaviour. The remaining problems consisted of social problems, attention problems, and thought problems. The latter were divided into psychotic and non psychotic items. A total of 14.3% of the adolescents were non-right-handed. We observed positive associations of non-right-handedness with thought problems, social problems, and being withdrawn and depressed. Externalising problems showed no associations with handedness. Within the thought problems subscale, the effect sizes associated with non-right-handedness for psychotic and non-psychotic items were 0.18 (p = .005) and 0.04 (p = .459), respectively. In conclusion, non-right handedness is predominantly associated with psychosis-related mental problems as early as in adolescence. Handedness could be taken into account when identifying adolescents at risk for psychosis. PMID- 19280380 TI - One membrane protein, two structures and six environments: a comparative molecular dynamics simulation study of the bacterial outer membrane protein PagP. AB - PagP is a bacterial outer membrane protein consisting of an 8 stranded transmembrane beta-barrel and an N-terminal alpha-helix. It is an enzyme which catalyses transfer of a palmitoyl chain from a phospholipid to lipid A. Molecular dynamics simulations have been used to compare the dynamic behaviour in simulations starting from two different structures (X-ray vs. NMR) and in six different environments (detergent micelles formed by dodecyl phosphocholine and by octyl glucoside, vs. four species of phospholipid bilayer). Analysis of interactions between the protein and its environment reveals the role played by the N-terminal alpha-helix, which interacts with the lipid headgroups to lock the PagP molecule into the bilayer. The PagP beta-barrel adopts a tilted orientation in lipid bilayers, facilitating access of lipid tails into the mouth of the central binding pocket. In simulations starting from the X-ray structure in lipid bilayer, the L1 and L2 loops move towards one another, leading to the formation of a putative active site by residues H33, D76 and S77 coming closer together. PMID- 19280381 TI - Polyethylenimine of various molecular weights as adjuvant for transfection mediated by cationic liposomes. AB - Over the last years significant progress has been made in non-viral gene delivery mediated by cationic liposomes. However, the results obtained are still far from being satisfactory regarding transfection efficiency, particularly when compared to that achieved using viral vectors. We have previously demonstrated that association of transferrin with cationic liposomes significantly improves transfection in a large variety of cells, both in vitro and in vivo. In this work, several strategies have been explored in order to further improve transfection mediated by transferrin-associated lipoplexes. To this regard, the effect on transfection of pre-condensation of DNA with polyethylenimine of low MWs (2.7, 2.0 and 0.8 KDa) at various N/P ratios, lipid composition, cationic lipid/DNA (+/-) charge ratio and the presence of a surfactant in the lipoplexes was investigated. Two different modes for preparing the liposomes were tested and the extent of cell association of their complexes with DNA as well as their capacity to protect the carried DNA were evaluated. Our results show that complexes generated from cationic liposomes prepared by the ethanol injection method in which the carried DNA was pre-condensed with low MW polyethylenimine are highly efficient in mediating transfection. The differential modulating effect observed upon association of transferrin to various liposome formulations on transfection mediated by the polyethylenimine-complexes suggests that these complexes enter into the cells through different pathways (involving clathrin versus caveolin), most likely by taking advantage of their intrinsic biophysical properties to escape from the endosome to the cytosol. PMID- 19280382 TI - Current concerns in Icelandic psychiatry; nation in crisis. PMID- 19280383 TI - In vitro stability and metabolism of salvinorin A in rat plasma. AB - Salvinorin A is the main active psychoactive ingredient in Salvia divinorum, a Mexican plant that has been widely available as a hallucinogen in recent years. The aims of this study were to investigate the stability of salvinorin A in rat plasma, esterases responsible for its degradation, and estimation of the degradation products. The apparent first-order rate constants of salvinorin A at 37 degrees C, 25 degrees C, and 4 degrees C were 3.8 x 10(-1), 1.1 x 10(-1), and < 6.0 x 10(-3) h(-1), respectively. Salvinorin A degradation was markedly inhibited by the addition of sodium fluoride, an esterase inhibitor. Moreover, phenylmethylsulfonyl fluoride (serine esterase inhibitor) and bis-p nitrophenylphosphate (carboxylesterase inhibitor) also inhibited salvinorin A degradation. In contrast, little or no suppression of the degradation was seen with 5,5'-dithiobis-2-nitrobenzoic acid (arylesterase inhibitor),ethopropazine (butyrylcholinesterase inhibitor), and BW284c51 (acetylcholineseterase inhibitor). These findings indicated that carboxylesterase was mainly involved in the salvinorin A hydrolysis in rat plasma.4. The degradation products of salvinorin A estimated by liquid chromatography-mass spectrometry included the deacetylated form (salvinorin B) and the lactone-ring-open forms of salvinorin A and salvinorin B. This lactone-ring-opening reactions were involved in calcium dependent lactonase. PMID- 19280384 TI - Assessment of cryopreserved human hepatocytes as a model system to investigate sulfation and glucuronidation and to evaluate inhibitors of drug conjugation. AB - Cultured cryopreserved human hepatocytes are extensively used as a model system for studying drug metabolism, although they remain poorly characterized in respect of the major conjugation reactions glucuronidation and sulfation. Using paracetamol (acetaminophen), we assessed eleven samples of cryopreserved human hepatocytes for their suitability to investigate the simultaneous glucuronidation and sulfation of xenobiotics and evaluated inhibitors of conjugation. Kinetic characterization showed broadly similar values for paracetamol conjugation by hepatocytes (as reported in the literature for in vitro systems), with Km values of approximately 6 mM and 0.3 mM for glucuronidation and sulfation, respectively. Substantial interindividual differences were observed. The hepatocytes demonstrated a strong dose-dependent switch from a preponderance of sulfation at low concentrations of paracetamol to glucuronidation at higher doses, consistent with routes of clearance in vivo. A number of drugs, some of which such as probenecid and sulfinpyrazone are known to interact with paracetamol in vivo, were demonstrated to inhibit the sulfation and/or glucuronidation of paracetamol in hepatocytes, demonstrating the potential application of this model system for studying drug-drug interactions involving conjugation. PMID- 19280385 TI - Putting sexuality (back) into HIV/AIDS: issues, theory and practice. AB - After more than twenty years of programming and activism aimed at stemming the sexual transmission of HIV (and addressing the needs of those most vulnerable to infection) the HIV/AIDS epidemic continues to grow worldwide. Taking up this concern, this paper argues that one of the reasons why HIV prevention has had limited success is because of inadequate conceptualization of human sexuality in such work. Giving sexuality a more prominent position in responses to the epidemic raises a range of issues, including theorization of gender, understanding of sexual subjectivity, the significance of pleasure (or lack of pleasure) in sexual decision-making, and conceptualization of sexual behaviour and culture. Taking these themes forward entails asking significant questions about the underlying paradigmatic and methodological commitments of mainstream HIV/AIDS research, especially the tendency to reproduce accounts of human sexuality as if it were a measurable form of conduct only. Advocating new approaches that take the meaning and symbolic value of sexualities into account complicates established orthodoxies in the field whilst offering potential for more effective HIV prevention strategies. PMID- 19280386 TI - The impact of a small steady stream of income for women on family health and economic well-being. AB - Our primary aim to evaluate the impact of a small steady stream of income on family health and well-being among rural women employed part-time in a health project in Sarlahi district, Nepal. All 870 women applying for the job of distributing nutritional supplements in their villages completed a questionnaire prior to selection for employment, 350 of whom were hired and 520 who were not. A total of 736 women completed a second questionnaire 2 years later, 341 (97.4%) of whom had been continuously employed during this period, and 395 (76.0%) who had never been employed by the project. Changes in health and well-being over 2 years were compared between women who were and were not hired. Women who were hired were younger and better educated, but were similar in other regards. After adjusting for selection differences, employed women were more likely to save cash, buy jewellery, and buy certain discretionary household goods over 2 years than those who were not hired. Expenditures on children's clothing increased more for employed women, and their children were more likely to be in private schools at follow-up, but there was no impact on health and survival of children. Women with a small steady stream of income did improve their personal economic situation by savings and increased expenditures for children and the household. Longer follow-up may reveal impacts on health access and expenditures, although these were not evident in 2 years of employment. PMID- 19280387 TI - Science and technology for wealth and health in developing countries. AB - It is now widely accepted that the developing world needs to invest in science and technology or risk falling behind as the technology gap between the North and South widens. However, these investments must be balanced by continued investment in basic population-wide services, such as healthcare and water supply and sanitation. Achieving this balance is a matter of ongoing debate in policy circles, and leaders and policy-makers in developing countries often have to make difficult decisions that pit investment in new technologies and capacity-building in science and technology against basic population-wide services such as healthcare and water supply and sanitation. The tension is underscored by evidence which suggests that rapidly industrializing economies, like in China, India and Brazil, are actually experiencing a rise in economic and health disparities among their populations. The fact that poor people in an industrializing country must fall behind while the rest of the country marches ahead does not have to be an inevitable outcome of industrialization. This article shows that science and technology can make an important and vital contribution to development, using public health as an example. It suggests the need to focus investments in science and technology in such a way that they can have a positive impact on public health. For instance, the use of simple, hand held molecular diagnostic tools can help unskilled health workers rapidly and accurately diagnose diseases, thus helping to reduce healthcare costs due to delayed or incorrect diagnoses. Recombinant vaccines can mitigate the risk of infection associated with live or attenuated vaccines, while needle-less delivery methods can help contain the spread of blood-borne infections. Critical to making technology investments work for population health are government policies and strategies that align public health goals and technology priorities. Such policies can include cross-sectoral training programs to improve dialogue between the technology and health sectors, setting up technology transfer cells to increase commercialization of health research relevant to local needs, and leveraging the phenomenon of low-margin high-volume marketing for health products. PMID- 19280388 TI - Experiences of women seeking medical care for obstetric fistula in Eritrea: implications for prevention, treatment, and social reintegration. AB - This article presents findings from qualitative interviews with women seeking medical care for obstetric fistula in Eritrea. The interviews were designed to inform programme design for the prevention and treatment of obstetric fistula. Interviews were conducted with 11 new fistula repair patients, 15 women returning for follow-up for their fistula repairs, and five accompanying family members at Massawa Hospital in the Northern Red Sea Zone of Eritrea during November-December 2004. The women described long delays in accessing emergency obstetric care due to delayed recognition of the seriousness of the problem and lack of transportation from remote villages. Follow-up patients described improvements in their conditions, but many continued to have problems with incontinence and sexual health. Both new and returning patients lacked specific information about their condition, what to expect in terms of treatment and recovery, and how to care for themselves. The findings point to a need for community mobilization and education on safe motherhood for prevention of fistula, as well as for improved information, counselling, follow-up, and social services for women who develop obstetric fistulas. PMID- 19280389 TI - Rethinking equal access: agency, quality, and norms. AB - In 2005 the Global Health Council convened healthcare providers, community organizers, policymakers and researchers at Health Systems: Putting Pieces Together to discuss health from a systems perspective. Its report and others have established healthcare access and quality as two of the most important issues in health policy today. Still, there is little agreement about what equal access and quality mean for health system development. At the philosophical level, few have sought to understand why differences in healthcare quality are morally so troubling. While there has been considerable work in medical ethics on equal access, these efforts have neglected health agency (individuals' ability to work toward health goals they value) and health norms, both of which influence individuals' ability to be healthy. This paper argues for rethinking equal access in terms of an alternative ethical aim: to ensure the social conditions in which all individuals have the capability to be healthy. This perspective requires that we examine injustices not just by the level of healthcare resources, but by the: (1) quality of those resources and their capacity to enable effective health functioning; (2) extent to which society supports health agency so that individuals can convert healthcare resources into health functioning; and (3) nature of health norms, which affect individuals' efforts to achieve functioning. PMID- 19280393 TI - As the world holds a vast array of cultures and circumstances, there is no one single 'right' approach to improving global health and equity. Foreword. PMID- 19280394 TI - Use of the non-pneumatic anti-shock garment (NASG) to reduce blood loss and time to recovery from shock for women with obstetric haemorrhage in Egypt. AB - Obstetric haemorrhage is one of the leading causes of maternal mortality. In many low-resource settings, delays in transport to referral facilities and in obtaining lifesaving treatment, contribute to maternal deaths. The non-pneumatic anti-shock garment (NASG) is a low-technology pressure device that decreases blood loss, restores vital signs, and has the potential to improve adverse outcomes by helping women survive delays in receiving adequate emergency obstetric care. With brief training, even individuals without medical backgrounds can apply this first-aid device. In this secondary analysis of hospital data from a pre-post intervention study in Egypt (N=364 women with obstetric haemorrhage and shock), 158 received standard care, while 206 received standard care plus the NASG. The NASG significantly reduced blood loss, time to recovery from shock, and, for those with postpartum haemorrhage due to uterine atony who received oxytocin, the NASG had a significant effect on blood loss independent of oxytocin. These results indicate that the NASG may be a valuable innovation for reducing maternal mortality in low-resource settings. Testing at community and household levels will be necessary in order to determine whether the NASG can help women survive the longest delays. PMID- 19280395 TI - HIV prevention for injection drug users in China and Vietnam: policy and research considerations. AB - A pattern of serious injection drug user (IDU) driven HIV epidemics in Asia, with emerging evidence of generalization through heterosexual transmission, indicates the need for interventions focusing on both drug- and sex-related risk reduction. In a cross-border HIV prevention project for IDUs in northern Vietnam and southern China, peer educators disseminated risk reduction information to IDUs in the community and provided 20,000-25,000 sterile needles/syringes and 4,000-6,000 condoms per month. Since implementation of these interventions, the frequency of both injecting and sexual risk behaviours fell significantly, HIV prevalence among IDUs declined or stabilized, and HIV incidence dropped. There is official support for harm reduction interventions in both countries but this appears precarious in view of persistently powerful political and financial support for a law enforcement approach. Moreover, the simultaneous pursuit of inconsistent policies can have negative effects on the implementation of interventions. A harmonized and consistent policy environment is needed. Most of the evidence for efficacy of community-based HIV prevention comes from the developed world, but well-designed evaluations of such interventions in Asia and elsewhere in the developing world would have a better chance to influence policy decisions there. A synergistic approach to research, policy development, and service delivery is best calculated to achieve positive results in the struggle against HIV/AIDS in developing countries. PMID- 19280396 TI - Facilitating interaction between TB and AIDS medical services for better management of patients with co-infections. AB - The Newly Independent States inherited two vertical services for tuberculosis (TB) and AIDS control from the Soviet Union. TB is an important cause of morbidity in the Central Asia Region (CAR). In recent years, HIV epidemics started growing rapidly, especially among injection drug users. TB is the most common AIDS-related opportunistic infection, leading to early deaths among AIDS patients and increased transmission of TB in the general population. An assessment carried out between March 2005 and May 2005 revealed that TB and AIDS services in CAR rarely interacted for the management of patients with co infections. Following the assessment, the USAID-funded CAPACITY Project promoted cooperation between TB and AIDS services in Uzbekistan, Tajikistan, and Kyrgyzstan. For the first time, representatives from the Ministries of Health and Justice, national and regional TB and AIDS centres, international organizations, and local non-governmental organizations (NGO) gathered to discuss mechanisms for linking TB and AIDS services to address the growing needs of co-infected patients. Technical Working Groups (TWG) established through this cooperation developed linkages between TB and AIDS services. These groups developed protocols, guidelines, and training programmes. Hundreds of medical specialists, health managers, and monitoring and evaluation specialists were trained and national teams of trainers were built. TB/HIV model sites were designed in each country and the models were implemented to test and refine the approaches for patients with co-infections. Key to the success of the approach has been the emphasis on wide participation from all stakeholders. Upon completion, successful models will be advocated for funding and national scale-up. PMID- 19280397 TI - Power, pleasure, pain, and shame: assimilating gender and sexuality into community-centred reproductive health and HIV prevention programmes in India. AB - Inspired by the vision of the Millennium Declaration, CARE and ICRW (International Centre for Research on Women) partnered with the Inner Spaces, Outer Faces Initiative (ISOFI) to learn how to more effectively integrate gender and sexuality into CARE's sexual and reproductive health programmes. Drawing from lessons learned from gender mainstreaming, ISOFI focuses initially on fostering personal change among staff, helping them to explore their own gender and sexuality 'baggage' and supporting transformation of their 'inner space'. ISOFI then gradually integrates mechanisms to promote organizational change, and finally extends to community development practice, the 'outer face'. As a system promoting change in organizational culture and practice, ISOFI features structured iterative loops of reflection and learning, action and experimentation, and analysis and assimilation. This article describes the ISOFI Innovation System, and reports on ISOFI-generated learning and innovation in sex positive HIV prevention programming for truckers and reproductive health interventions for women in India. PMID- 19280398 TI - Finally, daddies in the delivery room: parents' education in Georgia. AB - In recent decades there has been an increasing trend towards male partners attending childbirth in post-industrialized countries. However, in some countries, due to limited cultural acceptability and outdated delivery practices, fathers are not welcomed or even allowed into the delivery room. World experience postulates that men's active participation and assistance during childbirth accelerates the delivery process and improves delivery outcomes. Fathers' involvement also promotes positive feelings about the birth experience, fosters paternal role attainment, and strengthens family bonding. Despite documented advantages of fathers' involvement in pregnancy and childbirth, it still remains challenging to implement. The Healthy Women in Georgia (HWG) programme, promoting family-centred maternity care in Georgia, has demonstrated the efficiency of childbirth educational classes combined with modernized labour and delivery practices as a means of increasing fathers' participation in pregnancy care and childbirth. Moreover, HWG interventions have shown that fathers can effectively provide early skin-to-skin contact to caesarean section delivered infants, contributing to the heat conservation of the babies and minimizing the likelihood of hypothermia. Advanced labour and delivery practices, adequate antenatal education, and father's increased participation in pregnancy care and delivery promoted by the HWG programme has been shown to improve delivery outcomes, ultimately leading to better maternal and child health. PMID- 19280399 TI - Pharmacovigilance and new essential drugs in Africa: Zambia draws lessons from its own experiences and beyond. AB - Jolted into action by the thalidomide tragedy, developed Western countries began to establish national systems for identifying and responding to adverse drug reactions and events (or pharmacovigilance systems) about 40 years ago. These systems focus on side effects, adverse reactions, and drug interactions. In developing countries, especially in Africa, the scope for pharmacovigilance needs to be broader (despite the additional challenges this brings) because of growing problems with substandard and counterfeit drugs and the need to have an early warning signal system for the development of antimicrobial resistance to the 'new essential drugs' that are barely beyond the clinical trial stage in Africa, e.g. artemisinin-combination therapy (ACT) for malaria and antiretrovirals (ARV) for HIV/AIDS. Zambia learned important lessons from its own initial experiences in attempting to use ACT as a pathfinder for pharmacovigilance, as well as its experience with other drug information systems. In preparing its own renewed plans, it also drew lessons from international experience, including the weaknesses of the Food and Drug Administration's approach to pharmacovigilance in the USA, the UK's 'yellow card scheme', Brazil's fledgling pharmacovigilance systems for AIDS treatment, and the guidance provided by the World Health Organization and the Uppsala Monitoring Centre. These lessons are relevant for other African countries and even for developed countries seeking to improve pharmacovigilance systems. PMID- 19280403 TI - Psychometric evaluation of the Trauma Symptoms Checklist for Children (TSCC) among children affected by HIV/AIDS in China. AB - The current study, utilizing the baseline data from a longitudinal assessment of psychosocial needs of children orphaned by HIV/AIDS or living with HIV-infected parents in China, was designed to assess the psychometric properties of the Trauma Symptoms Checklist for Children (TSCC) among children affected by HIV/AIDS in China. The psychometric properties assessed in this study include internal consistency, convergent and discriminant validity, and construct validity. The sample in the current study include 296 double orphans, 459 single orphans, and 466 children living with HIV-infected parents in central China where many residents had been infected with HIV through unhygienic blood collection. The results demonstrate adequate reliability and validity of the TSCC among study population. Children who experienced more traumatic events scored significantly higher on all TSCC clinical scales and subscales than those children who experienced less such events. The Chinese version of the TSCC should provide mental health researchers and practitioners with a reliable and valid assessment instrument in studying posttraumatic distress and related psychological symptomology among children affected by HIV/AIDS in China. PMID- 19280404 TI - Quality of life and social support among patients receiving antiretroviral therapy in Western Uganda. AB - Quality of life (QOL) among patients with HIV/AIDS has been shown to improve once treatment with antiretroviral therapy (ART) has been initiated. We conducted a cross-sectional study in Western Uganda to examine the factors associated with QOL among patients who had received ART for the duration of at least six months. We interviewed 330 patients attending the HIV/AIDS clinic at two government supported hospitals in Western Uganda. We measured QOL using a culturally adapted version of the Medical Outcomes Study (MOS-HIV) tool and calculated the physical health summary (PHS) and mental health summary (MHS) scores. In addition, data were collected on sociodemographic factors, three-day self-reported adherence, social support, sexual behavior, CD4 count and viral load. Informational social support was significantly positively correlated with PHS (p=0.001) and MHS (p=0.002). Affectionate support was also significantly positively correlated to PHS (p=0.05) and MHS (p=0.03) but tangible support was not (PHS p value=0.85 and MHS p value=0.31). In the univariate analysis, older age, rural dwelling, alcohol use, CD4 count less than 200, and ART duration of less than one year were significantly associated with lower PHS scores. Lower PHS scores were also associated with sexual inactivity. In multivariate analysis, higher scores on informational social support and CD4> or =200 were associated with higher PHS score and past or recent alcohol consumption was associated with lower scores on MHS. Optimizing ART to restore CD4 count and provision of informational and affectionate social support but not tangible support, to HIV/AIDS patients may improve their QOL. PMID- 19280405 TI - Which factors hinder the decision of Polish HIV-positive patients to take up antiretroviral therapy? AB - The implementation of highly active antiretroviral therapy (HAART) in 1996 has significantly reduced mortality and morbidity for HIV-positive patients worldwide. However not all eligible persons start HAART. To identify reasons for therapy refusal by HIV-positive persons we performed a questionnaire study. The investigation was conducted among 321 HIV-positive individuals and focused on the decision to take up antiretroviral treatment. Out of 71 untreated patients, 34 (47.9%) admitted that in their case the therapy was not indicated, whereas 20 (28.3%) were afraid of potential side effects that might change their appearance, e.g. face lipoatrophy. Only the treated patients had been prepared to take up therapy, although 17 patients (6.0%) had not received any explanation of the therapy principles, aims or necessity to comply with medication regime. The therapy is generally not discussed with the patients for whom it is not currently indicated, which may contribute to the fixation of fears and prejudices. Doctors who treat HIV-positive patients should be aware of the prejudices and fears their patients have towards antiretroviral therapy in order to react properly and by means of the available antiretroviral drugs help prolong life and improve its quality. PMID- 19280406 TI - Profile of adults seeking voluntary HIV testing and counseling in rural Central India: results from a hospital-based study. AB - Rural India has an undetected load of HIV-positive individuals. Few rural adults present for HIV testing and counseling due to stigma, discrimination, and fear of social ostracization. In this rural hospital clinic-based study, we document profiles of rural adults seeking voluntary testing and counseling, and analyze correlates of HIV seropositivity. This cross-sectional study was conducted in 450 participants presenting to the outpatient clinics of Mahatma Gandhi Institute of Medical Sciences, Sevagram, Central India. After informed consent, pre- and post test counseling, HIV testing, and face-to-face interviews were conducted. Data were collected using a structured questionnaire. The median age of the 450 study participants was 34 years (range 18-88 years); the majority (74%) was married. The overall proportion of HIV seropositivity was 32% [95% CI 28%, 37%]. The proportions of HIV seropositivity in married women, married men, and single men were 41%, 37%, 18%, respectively. No single woman was found seropositive in the study. Very few married women were aware of their husbands' HIV status. In a multivariate analysis, correlates of HIV seropositivity in men were: age 30-39 years, being married, having sex with multiple partners, use of alcohol before sex, and testing positive for HIV in the past. In married women, the only predictor of seropositivity was being married. Although limited by the non-random nature of the sampling method, this pilot study is unique in that it is the first from this rural region of Central India. It provides baseline data on marginalized, largely unstudied populations that may aid in designing probabilistic community-based surveys in this neglected population. PMID- 19280407 TI - Relational contexts in adjustment to pregnancy of HIV-positive women: relationships, social support and personal adjustment. AB - Relational contexts are a central issue in most people's lives, and people usually rely on the support of others in everyday circumstances. Social support, as a relational context, could have a positive influence on personal adjustment, and is particularly relevant in the psychological well-being of HIV patients. Guided by the Convoy Model of social networks, in a sample of 31 HIV-positive pregnant women we try to assess the role of social support and social network in the adjustment to pregnancy. Profile analysis suggests a greater importance of social support provided by the partner and both parents, especially the support provided by the mother. At the same time, it seems to highlight the buffering hypothesis of social support, which could be understood as a protection factor in the adjustment of HIV-infected women' to pregnancy. PMID- 19280408 TI - Experiencing uncertain HIV treatment delivery in a transitional setting: qualitative study. AB - BACKGROUND: Advances in HIV treatment availability mean that the promise of highly active anti-retroviral treatment to turn HIV into a manageable chronic illness is becoming a reality for millions. However the mutability of the virus means that treatment adherence demands are high, and the supply of these life saving treatments needs to be constant. The onus is generally placed on the individual to adhere, and there is little focus in research or policy on the state's adherence to delivering treatment consistently. METHODS: We undertook in depth qualitative interviews to explore the narratives of HIV treatment experience among 41 people living with HIV (PLHIV) infection and 18 HIV treatment service providers in Serbia and Montenegro, a transitional setting in which state delivered and funded HIV treatment is inconsistently available. Data were analysed inductively and thematically. FINDINGS: Treatment shortages were common so the delivery of appropriate HIV treatment was not continuous. Access to reliable treatment and supply forecast information was weak and uneven. The insecure treatment situation fostered significant anxiety amongst PLHIV. INTERPRETATION: In the absence of reliable and sustained treatment access, information and support, PLHIV absorb the anxieties of system failures. This uncertainty led to an individuation of "treatment". PLHIV adopted rationing strategies to mediate their anxiety, energy and hope. This predominately resulted in varying forms of disengagement and neglect for social change. It is likely that this has significant negative implications for the promotion of HIV treatment advocacy and anti-stigma efforts. PMID- 19280409 TI - Does "asymptomatic" mean without symptoms for those living with HIV infection? AB - Throughout the history of the HIV epidemic, HIV-positive patients with relatively high CD4 counts and no clinical features of opportunistic infections have been classified as "asymptomatic" by definition and treatment guidelines. This classification, however, does not take into consideration the array of symptoms that an HIV-positive person can experience long before progressing to AIDS. This short report describes two international multi-site studies conducted in 2003 2005 and 2005-2007. The results from the studies show that HIV-positive people may experience symptoms throughout the trajectory of their disease, regardless of CD4 count or classification. Providers should discuss symptoms and symptom management with their clients at all stages of the disease. PMID- 19280410 TI - Hopes, fears, knowledge and misunderstandings: responses of HIV-positive mothers to early knowledge of the status of their baby. AB - Little is known about how HIV-positive mothers experience and react to knowing the HIV status of their baby as diagnosed by the polymerase chain reaction (PCR) test at 4-6 weeks. This qualitative study drew on interviews with 20 mothers of HIV-negative and 18 mothers of HIV-positive babies after receiving their baby's PCR results. Thematic analysis combined exploration of themes that appeared significant to the participants and those relevant to health care. Amongst the themes identified were the following: The period before getting the results involved active mental preparation and was emotionally stressful. Most women accepted the results, but some had doubts about their reliability. Mothers of HIV negative babies were relieved, but mothers of HIV-positive babies were generally very distressed and expressed a sense of responsibility and guilt. Both groups of mothers had similar hopes for the future of their babies, but the timelines of mothers of HIV-positive babies tended to be shorter. Most women experienced significant levels of stress, but were able to call on support networks and use various individual coping mechanisms to manage their stress. Most women were formula feeding their babies, but regretted not being able to breastfeed. Many women had not planned their current baby and most did not intend to have more children, but many of the latter had not taken active steps to prevent further pregnancy. The findings provide pointers to shortcomings in health worker communication and suggest that more effective communication should take account of normative community views and be more closely attuned to the changing needs and experiences of HIV-positive mothers. PMID- 19280411 TI - Women, economic hardship and the path of survival: HIV/AIDS risk behavior among women receiving HIV/AIDS treatment in Uganda. AB - The results are presented from a 2005 survey of 377 women in four HIV/AIDS treatment programs in Uganda. The aim of the study was to explore women's economic hardships and the association with four sexual risk behaviors: whether a woman was sexually active in the last 12 months, whether a condom was used during the last sex act, whether she reported having had a sexual partner in the last six months who she suspected had multiple partners and report of forced, coercive or survival sex in the last six months. Few women were sexually active (34%), likely due to the high proportion of widows (49%). Married women were likely to report forced, coercive or survival sex (35%). Eighty-four percent of women reported condom used at last sex act. Forced, coercive or survival sex was associated with number of meals missed per week (AOR=1.125, 95% CI 1.11, 1.587, p<0.05). Sex with a partner in the last six months who a woman suspected had multiple partners was also associated with number of missed meals per week (AOR=2.080, 95% CI 1.084, 3.992). Currently women in Ugandan antiretroviral therapy programs are not likely to be sexually active, except for married women. Many women need to find food and other support, which may put them at risk of forced, coercive or survival sex due to dependency on men. PMID- 19280412 TI - The fork in the road: HIV as a potential positive turning point and the role of spirituality. AB - We interviewed 147 HIV-positive people regarding their key life-changing experiences - involving profound changes in attitudes, behaviors, beliefs (including spiritual beliefs), or self-views - to determine the prominence of HIV as the key positive/negative turning point. HIV was the key turning point, for 37% (26% positive, 11% negative), whereas for 63% of our sample it was not. Characteristics associated with perceiving HIV as the most positive turning point included having a near-death experience from HIV, increasing spirituality after HIV diagnosis, and feeling chosen by a Higher Powerto have HIV. Notably, perceived antecedents of viewing HIV as the key positive turning point were hitting rock bottom and calling on a Higher Power. Conversely, viewing HIV as the most negative turning point was associated with declining spirituality after diagnosis. Spirituality can both negatively and positively affect coping with HIV. Promoting positive spiritual coping may offer new counseling approaches. Further, for the majority of the participants, HIV is not the key turning point, which may be an indicator of the normalization of HIV with the advent of effective treatment. PMID- 19280413 TI - A community perspective on young people's knowledge of HIV/AIDS in three African countries. AB - Individual, household and community-level influences on young people's (15-24) knowledge of HIV/AIDS in three African countries (Burkina Faso, Ghana and Zambia) are explored. The focus of the analysis is on the roles of demographic, economic and behavioral dimensions of the community environment in shaping knowledge of HIV/AIDS. Data from Demographic and Health Surveys, collected independently for males and females in each of the countries, are analyzed. There are clear pathways through which the community environment shapes knowledge, and the community influences on knowledge vary greatly by country and gender. For young women, residences in communities with demographic and behavioral patterns that are indicative of greater opportunities are associated with increased knowledge of HIV/AIDS. The results highlight community-level factors that can be harnessed in the development of community-based interventions to improve knowledge of HIV/AIDS among young people, and reinforce the need to focus on the community environment in designing behavioral change interventions. PMID- 19280414 TI - Family relationships in people living with HIV in a city in the USA. AB - The purpose of this study was to identify relationships of individuals that people living with human immunodeficiency virus/acquired immune deficiency syndrome (PLWH) in a city in the USA include in their family of choice, or chosen family. The convenience sample in this cross-sectional, exploratory study consisted of 150, mostly male and African American PLWH. Participants included partners/lovers most frequently, followed by children, mother, and siblings, and grandparents in their chosen family. Family structures of PLWH in this study are consistent with the diverse nature of families within society. PMID- 19280415 TI - Discussing matters of sexual health with children: what issues relating to disclosure of parental HIV status reveal. AB - Little is published about the disclosure of parents' own HIV status to their children in Africa. Research shows that keeping family secrets from children, including those related to a parent's HIV status, can be detrimental to their psychological well-being and to the structure of the family. Further, children with HIV-positive parents have been shown to be more vulnerable to poorer reproductive health outcomes. This qualitative study in Botswana conducted in depth interviews among 21 HIV-positive parents on antiretroviral therapy. The data revealed that parents found discussing the issue of HIV with children difficult, including disclosing their own HIV status to them. Reasons for disclosing included: children being HIV positive, the rest of the family knowing, or the parent becoming very sick. Reasons for not disclosing included: believing the child to be too young, not knowing how to address the issue of HIV, that it would be "too painful" for the child/ren. Concern that other people might find out about their status or fear of children experiencing stigmatising behaviour. Interviews elucidated the difficulty that parents have in discussing their own HIV status and more general sexual health issues with their children. Parents and other guardians require support in managing age-appropriate disclosure to their children. This may further enable access to forums that can help children cope with their fears about the future and develop life skills in preparation for dealing with relationships of a sexual nature and sexual health as children move into adulthood. In developing such support mechanisms, changing family roles in Botswana need to be taken into consideration and the role of other family members in the upbringing of children in Tswana society need to be recognised and utilised. PMID- 19280418 TI - Impact factors and the h-index: what researchers and readers need to know. PMID- 19280419 TI - Supplementary online material: potential and precautions. PMID- 19280420 TI - DECtalk and VeriVox: intelligibility, likeability, and rate preference differences for four listener groups. AB - The intelligibility, likeability, and speech presentation rates for two types of popular synthesized speech, DECtalk and VeriVox, were investigated across four age groups ranging from 6-85 years old. Results for all age groups revealed differences in intelligibility between synthesized adult and child voices, with intelligibility scores for child synthesized voices significantly below those for adult synthesized voices. The most popular synthesized voices were from the VeriVox family. Of interest is that preferred speech presentation rate, expressed as mean words per minute, increased as a function of age, with the oldest listeners preferring the fastest speech presentation rate. This is inconsistent with previous studies that have investigated rate preferences among older adults, and could reflect the different synthesized speech options employed in this study. Implications of these findings for the improvement of text-to-speech products and the practice of AAC are discussed, along with suggestions for further research. PMID- 19280421 TI - Comparison of three head-controlled mouse emulators in three light conditions. AB - Three infrared head-controlled mouse emulators (HeadMouse Extreme, Tracker Pro and Tracker Inside) were paired with an augmentative and alternative communication (AAC) device (Pathfinder) and tested for speed and accuracy in three different light conditions. Eleven participants tested the systems, entering a list of common statements used by AAC users. The Language Activity Monitor (LAM) recorded all selections made. The data were analyzed for error rate and selection rate. The results revealed that (a) in a single light condition, the different mouse emulators performed significantly differently from each other, (b) in general, a single mouse emulator performed consistently across all light conditions, and (c) the external mouse emulators performed better than the internal emulator, for speed and accuracy, in all light conditions. PMID- 19280422 TI - Recognition and use of line drawings by children with severe intellectual disabilities: the effects of color and outline shape. AB - Communication symbols for students with severe intellectual disabilities often take the form of computer-generated line drawings. This study investigated the effects of the match between color and shape of line drawings and the objects they represented on drawing recognition and use. The match or non-match between color and shape of the objects and drawings did not have an effect on participants' ability to match drawings to objects, or to use drawings to make choices. PMID- 19280423 TI - AAC technology transfer: an AAC-RERC report. AB - Transferring innovative technologies from the university to the manufacturing sector can often be an elusive and problematic process. The Rehabilitation and Engineering Research Center on Communication Enhancement (AAC-RERC) has worked with the manufacturing community for the last 10 years. The purpose of this article is to discuss barriers to technology transfer, to outline some technology transfer strategies, and to illustrate these strategies with AAC-RERC related activities. PMID- 19280424 TI - Adverse reactions associated with acetylcysteine. AB - INTRODUCTION: Paracetamol (acetaminophen) is one of the most common agents deliberately ingested in self-poisoning episodes and a leading cause of acute liver failure in the western world. Acetylcysteine is widely acknowledged as the antidote of choice for paracetamol poisoning, but its use is not without risk. Adverse reactions, often leading to treatment delay, are frequently associated with both intravenous and oral acetylcysteine and are a common source of concern among treating physicians. METHODS: A systematic literature review investigating the incidence, clinical features, and mechanisms of adverse effects associated with acetylcysteine. RESULTS: A variety of adverse reactions to acetylcysteine have been described ranging from nausea to death, most of the latter due to incorrect dosing. The pattern of reactions differs with oral and intravenous dosing, but reported frequency is at least as high with oral as intravenous. The reactions to the intravenous preparation result in similar clinical features to true anaphylaxis, including rash, pruritus, angioedema, bronchospasm, and rarely hypotension, but are caused by nonimmunological mechanisms. The precise nature of this reaction remains unclear. Histamine now seems to be an important mediator of the response, and there is evidence of variability in patient susceptibility, with females, and those with a history of asthma or atopy are particularly susceptible. Quantity of paracetamol ingestion, measured through serum paracetamol concentration, is also important as higher paracetamol concentrations protect patients against anaphylactoid effects. Most anaphylactoid reactions occur at the start of acetylcysteine treatment when concentrations are highest. Acetylcysteine also affects clotting factor activity, and this affects the interpretation of minor disturbances in the International Normalized Ratio in the context of paracetamol overdose. CONCLUSION: This review discusses the incidence, clinical features, underlying pathophysiological mechanisms, and treatment of adverse reactions to acetylcysteine and identifies particular "at-risk" patient groups. Given the commonality of adverse reactions associated with acetylcysteine, it is important to ensure that any adverse event does not preclude patients from receiving maximal hepatic protection, particularly in the context of significant paracetamol ingestion. Further work on mechanisms should allow specific therapies to be developed. PMID- 19280425 TI - Aluminium and zinc phosphide poisoning. AB - INTRODUCTION: Aluminium and zinc phosphides are highly effective insecticides and rodenticides and are used widely to protect grain in stores and during its transportation. Acute poisoning with these compounds may be direct due to ingestion of the salts or indirect from accidental inhalation of phosphine generated during their approved use. MECHANISMS OF TOXICITY: Both forms of poisoning are mediated by phosphine which has been thought to be toxic because it inhibits cytochrome c oxidase. While phosphine does inhibit cytochrome C oxidase in vitro, the inhibition is much less in vivo. It has been shown recently in nematodes that phosphine rapidly perturbs mitochondrial morphology, inhibits oxidative respiration by 70%, and causes a severe drop in mitochondrial membrane potential. This failure of cellular respiration is likely to be due to a mechanism other than inhibition of cytochrome C oxidase. In addition, phosphine and hydrogen peroxide can interact to form the highly reactive hydroxyl radical and phosphine also inhibits catalase and peroxidase; both mechanisms result in hydroxyl radical associated damage such as lipid peroxidation. The major lethal consequence of phosphide ingestion, profound circulatory collapse, is secondary to factors including direct effects on cardiac myocytes, fluid loss, and adrenal gland damage. In addition, phosphine and phosphides have corrosive actions. CLINICAL FEATURES: There is usually only a short interval between ingestion of phosphides and the appearance of systemic toxicity. Phosphine-induced impairment of myocardial contractility and fluid loss leads to circulatory failure, and critically, pulmonary edema supervenes, though whether this is a cardiogenic or non-cardiogenic is not always clear. Metabolic acidosis, or mixed metabolic acidosis and respiratory alkalosis, and acute renal failure are frequent. Other features include disseminated intravascular coagulation, hepatic necrosis and renal failure. There is conflicting evidence on the occurrence of magnesium disturbances. MANAGEMENT: There is no antidote to phosphine or metal phosphide poisoning and many patients die despite intensive care. Supportive measures are all that can be offered and should be implemented as required. PMID- 19280426 TI - Carnitine in the treatment of valproic acid-induced toxicity. AB - INTRODUCTION: Valproic acid (VPA) is a broad-spectrum antiepileptic drug that is now used commonly for several other neurological and psychiatric indications. VPA is usually well tolerated, but serious complications, including hepatotoxicity and hyperammonemic encephalopathy, may occur. These complications may also arise following acute VPA overdose, the incidence of which is increasing. Intoxication usually only results in mild central nervous system depression, but serious toxicity and death have been reported. VALPROIC ACID AND CARNITINE: As a branched chain carboxylic acid, VPA is extensively metabolized by the liver via glucuronic acid conjugation, mitochondrial beta- and cytosolic omega-oxidation to produce multiple metabolites, some of which may be involved in its toxicity. Carnitine is an amino acid derivative that is an essential cofactor in the beta-oxidation of fatty acids. It is synthesized endogenously from the essential amino acids, methionine and lysine. VPA inhibits the biosynthesis of carnitine by decreasing the concentration of alpha-ketoglutarate and may contribute to carnitine deficiency. It is postulated that carnitine supplementation may increase the beta oxidation of VPA, thereby limiting cytosolic omega-oxidation and the production of toxic metabolites that are involved in liver toxicity and ammonia accumulation. VPA-induced hepatotoxicity and hyperammonemic encephalopathy may be promoted either by a pre-existing carnitine deficiency or by deficiency induced by VPA per se. CARNITINE SUPPLEMENTATION: Some experimental and clinical data suggest that early intravenous supplementation with l-carnitine could improve survival in severe VPA-induced hepatotoxicity. Carnitine administration has been shown to speed the decrease of ammonemia in patients with VPA-induced encephalopathy although a correlation between ammonia concentrations and the clinical condition was not always observed. As it does not appear to be harmful, l-carnitine is commonly recommended in severe VPA poisoning, especially in children, although the clinical benefit in terms of liver protection or hastening of recovery from unconsciousness has not been established clearly. Prophylactic carnitine supplementation is also advocated during VPA therapy in high-risk pediatric patients. CONCLUSION: Further controlled, randomized, and probably multicenter trials are required to better delineate the therapeutic and prophylactic roles of l-carnitine and the optimal regimen of administration in the management of VPA toxicity. PMID- 19280427 TI - Late laryngeal findings in sulfur mustard poisoning. AB - INTRODUCTION: Sulfur mustard is an alkylating warfare agent and its inhalation results in early and late toxic effects. This agent was used widely against both military and civilian population by Iraqi forces in the Iran-Iraq war (1983-88). The purpose of the study was to describe abnormal laryngeal findings in a group of chemical war injury patients (soldiers) 20 years after an acute exposure to sulfur mustard. METHODS: In this observational case series study, 50 male patients who were suffering from chronic respiratory symptoms and because of exacerbation of these problems, were consecutively selected and clinically examined for dysphonia by a speech-language pathologist, and then underwent fiberoptic laryngobronchoscopy to describe anatomical and functional pathologic findings of the larynx. RESULTS: In objective speech evaluation, different degrees of dysphonia including harshness and hoarseness were observed in 46% of patients. Inflammation was present in supraglottic and subglottic regions of 9 (18%) and 3 (6%) patients, respectively. Hyperfunction of the false vocal cords was observed in 24 (48%) patients. CONCLUSION: Chronic laryngitis was seen in a considerable number of exposed victims nearly 20 years after exposure, in spite of different treatments received. PMID- 19280428 TI - Critical reviews in biochemistry and molecular biology. Introduction. PMID- 19280429 TI - Nuclear RNA surveillance: no sign of substrates tailing off. AB - The production of cellular RNAs is tightly regulated to ensure gene expression is limited to appropriate times and locations. Elimination of RNA can be rapid and programmed to quickly terminate gene expression, or can be used to purge old, damaged or inappropriately formed RNAs. It is elimination of RNAs through the action of a polyadenylation complex (TRAMP), first described in the yeast Saccharomyces cerevisiae, which is the focus of this review. The discovery of TRAMP and presence of orthologs in most eukaryotes, along with an increasing number of potential TRAMP substrates in the form of new small non-coding RNAs, many of which emanate from areas of genomes once thought transcriptionally silent; promise to make this area of research of great interest for the foreseeable future. PMID- 19280430 TI - SRA and its binding partners: an expanding role for RNA-binding coregulators in nuclear receptor-mediated gene regulation. AB - The discovery that SRA RNA can function as a nuclear receptor (NR) coactivator resulted in a fundamental change in the perception of how NRs and their coregulators may regulate hormone signaling pathways. The subsequent identification of molecules capable of binding SRA, including SHARP, p68, and more recently SLIRP, which also function as coregulators, has further broadened our understanding of NR-dependent gene regulation. The integral role that NRs play in directing developmental, metabolic and pathological programs of transcription has defined them as paramount targets for treating a broad range of human diseases. Thus with a greater understanding of SRA and its interactions with its binding partners, novel RNA-protein interactions may be identified and exploited for therapeutic gain. Here we discuss the isolation of SRA, its impact on NR activity and interactions with known binding partners. PMID- 19280431 TI - Riboactivators: transcription activation by noncoding RNA. AB - The paradigm of gene regulation was forever changed by the discovery that short RNA duplexes could directly regulate gene expression. Most regulatory roles attributed to noncoding RNA were often repressive. Recent observations are beginning to reveal that duplex RNA molecules can stimulate gene transcription. These RNA activators employ a wide array of mechanisms to up-regulate transcription of target genes, including functioning as DNA-tethered activation domains, as coactivators and modulators of general transcriptional machinery, and as regulators of other noncoding transcripts. The discoveries over the past few years defy "Moore's law" in the breath-taking rapidity with which new roles for noncoding RNA in gene expression are being revealed. As gene regulatory networks are reconstructed to accommodate the influence of noncoding RNAs, their importance in maintenance of cellular health will become increasingly apparent. In fact, a new generation of therapeutic agents will focus on modulating the function of noncoding RNA. PMID- 19280432 TI - Non-cancer health effects of diesel exhaust: a critical assessment of recent human and animal toxicological literature. AB - We reviewed laboratory and clinical studies bearing on the non-cancer health effects of diesel exhaust (DE) published since the 2002 release of the US EPA Health Assessment Document for Diesel Engine Exhaust. We critically evaluated over 100 published articles on experimental research, focusing on their value for predicting the risk of non-cancer health effects in humans exposed to DE. Human controlled-exposure studies provide new evidence of lung inflammatory effects and thrombogenic and ischemic effects of inhaled DE, albeit for older-model diesel engines and concentrations that are much higher (approximately 300 microg/m(3)) than typical ambient or even occupational levels. Recent animal studies provide insight into the potential mechanisms underlying observed respiratory and cardiovascular health responses; however, because of unrealistically high DE concentrations, the mechanisms elucidated in these studies may not be relevant at lower DE exposure levels. Although larger in number, and suggestive of possible mechanisms for non-cancer health effects at elevated DE levels, interpretation of this recent group of clinical-study findings and laboratory-animal results remains hindered by inconsistencies and variability in outcomes, potentially irrelevant DE-exposure compositions, limitations in exposure protocols and pathways, and uncertainties in extrapolation and generalization. A mechanism of action that allows reliable prediction of adverse health effects at DE-exposure levels typical of the present-day ambient and occupational environment has not emerged. Because of changing diesel-engine technology, inhalation studies using realistic environmental and occupational exposures of new-technology diesel exhaust are of critical importance. PMID- 19280434 TI - Psychosocial adjustment to multiple sclerosis: exploration of identity redefinition. AB - PURPOSE: As multiple sclerosis (MS) often occurs in the prime of life and is unpredictable in nature, there is likely to be a strong psychological effect, with changes in values and beliefs and how the individual sees him or her self. This article presents the findings of a focus group study which aimed to explore the subjective experiences of living with, and adjusting to, MS. METHOD: Seven individuals who had been diagnosed with MS for at least 5 years reflected on their reactions to being diagnosed, how they cope with the day to day challenges of the disease, and the changes that they have experienced. Data were transcribed verbatim and analysed using interpretative phenomenological analysis. RESULTS: Diagnosis was met with negative reactions: denial, concealment and diminished confidence. However, the majority reported that, over time, there were positive changes in terms of their values and outlook. It would appear that the functional difficulties and psychological challenges, such as uncertainty and depression, are ameliorated to some extent by an increased appreciation for life and spirituality. CONCLUSIONS: The findings provide insight into the psychological process of identity redefinition associated with adjusting to MS. Given this, interventions should target role/identity re-examination to assist individuals with MS in better managing the disease and enjoying life. PMID- 19280433 TI - The endocrine effects of mercury in humans and wildlife. AB - Mercury (Hg) is well studied and research continues as our knowledge of its health risks increases. One expanding area of research not well emphasized to date is the endocrine effects of Hg. This review summarizes the existing literature on the effects of Hg on the endocrine system and identifies gaps in the knowledge. It focuses on the thyroid, adrenal, and reproductive systems, including the accumulation of Hg in the endocrine system, sex differences that are manifested with Hg exposure, reproductive effects in male and female animals including humans, and Hg effects on the thyroid and adrenal systems. We concluded that there are five main endocrine-related mechanisms of Hg across these systems: (a) accumulation in the endocrine system; (b) specific cytotoxicity in endocrine tissues; (c) changes in hormone concentrations; (d) interactions with sex hormones; and (e) up-regulation or down-regulation of enzymes within the steroidogenesis pathway. Recommendations for key areas of research to better understand how the endocrine effects of Hg affect human and wildlife health were developed, and include increasing the amount of basic biological information available about Hg and wildlife species, exploring the role of Hg in the presence of other stressors and chemicals, understanding sublethal and indirect effects of Hg on adverse outcomes, developing better methods to extrapolate effects across species, and understanding the effects of Hg on multiple organ systems following exposure of an animal. Greater inclusion of endocrine endpoints in epidemiological and field studies on humans and wildlife will also advance the research in this area. PMID- 19280435 TI - Memory and executive function predict mobility rehabilitation outcome after lower limb amputation. AB - PURPOSE: Post-amputation rehabilitation is physically and cognitively demanding. Understanding which specific cognitive domains mediate outcome is critical to the development of interventions. METHOD: A cohort undergoing post-amputation rehabilitation was assessed before limb fitting and followed up at 6 months (n = 34). The average age was 60.69 years (SD = 13.98). 82.4% of the sample was male. 79.4% had amputations because of peripheral arterial disease. Memory, visuospatial function, executive function, praxis, emotion and language were assessed at Time 1 (first prosthetic clinic attendance). Time 1 data were also gathered on aetiology, level of amputation, comorbidities, pain and demographics. Six month outcomes were the locomotor capability index (LCI), the special interest group in amputee medicine (SIGAM) mobility grades and self reported hours of use. RESULTS: The LCI at 6 months was significantly predicted in regression analyses by a measure of visual memory (figure recall) (adjusted R2 = 24.8%, df = 32, zbeta = 0.52, p = 0.002. Hours of use were predicted by the verbal fluency test total (adjusted R2 = 17.1%, df= 26, zbeta = 0.45, p = 0.017). SIGAM mobility grades were predicted by a combination of immediate verbal memory (story recall), age, level of amputation and presence of pain (adjusted R2 = 58.2, df = 30, zbeta = 0.52, p = 0.000). CONCLUSIONS: Neuropsychological and clinical variables predict a large amount of 6 month outcome variance. Cognitive difficulties may be considered mediators of poor outcome. PMID- 19280436 TI - Factors explaining return to work for long-term sick workers in Norway. AB - PURPOSE: This study investigates factors that will affect the chances of long term sick workers returning to work. METHOD: The study is based on a representative sample of persons with mental illnesses and musculoskeletal disorders who received a rehabilitation allowance in Norway in 2002. Their occupational status in 2005 and experiences from the rehabilitation process were charted through their responses to a questionnaire in combination with national register data. Our analytical framework is based on three domains: the medical domain, the domain of authoritative bodies and the production domain (working life), as described by Lindqvist (Att satta granser: organisationer och reformer i arbetsrehabiliteringen. Umea: Borea; 2000). RESULTS: Our main findings are that earlier work experience, age and other members in household, in addition, to influence over one's own rehabilitation process explains whether workers on long term sick leave return to work. Furthermore, individual factors such as gender and diagnosis (i.e. musculoskeletal disorder and mental illness) do not explain the probability to return to work. Experience and contact with representatives of the medical, the authoritative bodies and the production domain only explain to a small extent the probability to return to work. It is even interesting to note that regular contact with the social insurance office (SIO) has a negative effect on the probability to return to work. This may indicate that contact with the SIO subjects sick workers to a risk of developing a 'social insurance career' to secure their income. CONCLUSIONS: The findings show that the medical, the authoritative bodies and the production domain might represent different logics that can pull a sick worker in different directions during the rehabilitation process. However, these different logics do not fully explain, which outcome a rehabilitation process takes. It is important to extend the analysis to include how the individuals respond to these logics during the rehabilitation process. PMID- 19280437 TI - Meaning perspective transformation following stroke: the process of change. AB - PURPOSE: Transformative Learning in an educational theory that posits that individuals learn and grow when their meaning perspectives (frames of reference for interpreting an experience based on knowledge, feelings, values and beliefs) are reformulated following a critical event. This theory has become quite influential in the exploration of adaptation to chronic illness. This study explored whether the change that occurs following stroke follows a process similar to transformative learning. METHOD: Grounded Theory approach was used to explore changes in meaning perspective among 12 people who were members of stroke support organisations, had a stroke at least 1 year prior to the study and described themselves as viewing life positively following stroke. Constant comparison analysis of interviews with these individuals was used to explore their experience following stroke. RESULTS: Meaning perspective transformation occurred with four factors contributing to transformation: triggers, support, knowledge and choices to action. A substantive grounded theory of the process of meaning perspective transformation following stroke is presented, which illustrates the interaction of these contributing factors in initiating and facilitating the transformation process. CONCLUSION: Transformative learning can offer insight into how people who have experienced stroke learn, rebuild competence and re-engage in valued activities. PMID- 19280438 TI - Encountering staff in the home: three older adults' experience during six months of home-based rehabilitation. AB - PURPOSE: The purpose of the present study was to explore and describe how older adults who received home-based rehabilitation perceived the staff during a period of 6 months when they received rehabilitation. Specifically, the study focused on how the participants collaborated with and made use of the services from the staff. METHOD: In this case-oriented study, three older adults were interviewed continuously during the 6-month period they received home-based rehabilitation. The interviews were analysed continuously using a grounded theory approach. RESULTS: Five different modes of perceiving the staff were identified among the participants: as small talk persons, as discussions partners, as instructors and advisors, as teachers and as persons who carry out tasks efficiently. The three conditions that most came to influence the way the participants perceived collaborated with the staff were as follows: 'experience and encounters with the staff', 'expectations for the future daily life', and 'the participants' needs and tasks related to their disability'. CONCLUSIONS: To achieve collaboration and user involvement, the staff have to encounter each client differently in accordance with the various tasks that must be carried out during rehabilitation. Our findings indicate that this is particularly important for older adults during home-based rehabilitation, since older adults often have changing needs because of comorbidity. PMID- 19280439 TI - Long-term ophthalmic health care in Usher syndrome type I from an ICF perspective. AB - PURPOSE: The aim was to explore ophthalmic health care in female patients with Usher Syndrome type I (USH I) over 20 years and to evaluate the relationship between the ophthalmic health care and the health state of the patients from a health perspective. METHODS: A retrospective study of records from ophthalmology departments (OD) and low vision clinics (LVC) from 1985 to 2004. Assessment of the reports was performed based on the International Classification of Functioning, Disability and Health (ICF). Findings were analysed by manifest content analysis with ICF as a framework and using four themes: health care system, procedure examinations, patient's functioning and disability and procedure actions. RESULTS: The records of nine female patients (aged 25-39 years, 1985) with USH I were selected from the national database of USH. A great number of notes were collected (OD 344 and LVC 566). Procedure examinations were exclusively oriented towards body structure and function. All patients showed aggravated visual impairment over and above the hearing and vestibular impairment. Procedure actions were oriented towards environmental factors. No correlation was found between procedures performed and patient's experience of disability. CONCLUSIONS: The high degree of resource allocation was not correlated to the patients' impairment. The study indicates that the ophthalmic health care was characterised by inefficiency. This conclusion is very serious because patients very likely face severe disability and emotional difficulties. ICF is ought to be incorporated in ophthalmic health care strategy to improve the health care. PMID- 19280440 TI - Validity, reliability and predictive accuracy of the Driving Awareness Questionnaire. AB - PURPOSE: To examine the psychometric properties of the Driving Awareness Questionnaire (DriveAware) and to compare this rating of awareness of driving ability with the rating made in the context of an on-road assessment. METHOD: A prospective design was employed to examine the psychometric properties of DriveAware using Rasch analysis. Participants (n = 91) with varying diagnoses were recruited from two driving rehabilitation centres in Sydney, Australia. They completed DriveAware in the context of a standard driving assessment. Awareness of driving ability determined by DriveAware was compared with awareness determined in the on-road assessment. RESULTS: Rasch analysis provided good evidence for construct validity and inter-rater reliability and some evidence for internal reliability of DriveAware. The item hierarchy was logical and goodness of-fit statistics for four of five items were within an acceptable range. The test had a moderate reliability index (0.73). Measurement precision could be improved by including more items to assess drivers with little awareness. When compared with the on-road measure of awareness, the best DriveAware cut-off score yielded sensitivity of 84% and specificity of 94%. CONCLUSION: This short and easy-to-administer test could provide a useful tool with sound psychometric properties to measure awareness of driving ability. PMID- 19280441 TI - A comparison of motor abilities and perceived self-efficacy between children with hearing impairments and normal hearing children. AB - PURPOSE: The present study evaluates the differences in motor abilities and in self-efficacy between children with hearing impairments and children with normal hearing. METHOD: Participants were 48 children, aged 5 - 9 years, including 22 with hearing impairments and 26 with normal hearing. Motor abilities were evaluated by the Children Activity Scales for Teachers (ChAS-T) and the Movement Assessment Battery for Children (MABC). Self-efficacy was evaluated using the Perceived Efficacy and Goal Setting System (PEGS). RESULTS: Children with hearing impairments showed significantly lower motor abilities, according to the ChAS-T and the MABC. No significant differences were found between the groups in regard to the self-efficacy level according to most evaluated scales. Results for the study group revealed a significant correlation between the ChAS-T and the static balance score of the MABC. In both groups, significant correlations were found between motor abilities and child's age. CONCLUSIONS: While children with hearing impairments may show lower motor abilities, as expressed by the MABC and the ChAS T, their self-efficacy is similar to that of their normal hearing peers. PMID- 19280442 TI - Statement of retraction. PMID- 19280443 TI - Why many visible minority women in Canada do not participate in cervical cancer screening. AB - OBJECTIVE: To determine a high-risk group of visible minority women in Canada who do not participate in cervical cancer screening and the reasons why they do not participate. DESIGN: We combined two cycles of a large Canadian health survey, Canadian Community Health Survey (CCHS), to obtain a large sample size of visible minority women. Proportions of 'never having a Papanicalaou (Pap) test' and 'not having a Pap test within the last three years' were then calculated for different ethnic groups using sampling weights advised by Statistics Canada to account for the complex sampling procedure used in CCHS. A logistic regression model was developed to test the association between demographic and health-related variables and not having a Pap test. To identify visible minority women who were at a high risk of not having a Pap test, we stratified these women simultaneously on three variables that were significant in the logistic regression model. RESULTS: Visible minority women were more than twice as likely never to have had a Pap test. Among visible minority women, those who recently immigrated to Canada and did not have a regular physician had the highest risk for not having a Pap test. Common reasons reported for not having a Pap test included believing it was not necessary and simply not getting around to it. CONCLUSION: Visible minority women in Canada may not be participating in regular Pap testing because of cultural beliefs and a lack of an understanding of the importance of Pap testing. A culturally appropriate cervical cancer screening intervention program that involves members of visible minority communities may increase participation of this subgroup of Canadian women. This study provides preliminary information on why visible minority women in Canada do not participate in cervical cancer screening. However, the lumping together of all visible minority may obscure differences between different ethnic groups. Therefore, further research on each ethnic group is required to develop tailored culturally appropriate intervention. PMID- 19280444 TI - Treatment-seeking behaviour among the Nasioi people of Bougainville: choosing between traditional and western medicine. AB - BACKGROUND: In Papua New Guinea (PNG) there continues to be considerable interest in developing a health system that incorporates both traditional and western medicine. A policy on traditional medicine has recently been endorsed. Simultaneously, there is limited information about the traditional beliefs and practices that influence treatment-seeking behaviour. AIM OF THE STUDY: A case study among the Nasioi people of Bougainville was conducted to gather information that could help to inform the implementation of the National Policy on Traditional Medicine for PNG. RESEARCH OBJECTIVES: The main objective of the case study was to describe how health knowledge and belief systems influence treatment seeking behaviour, specifically in relation to the use of traditional and western health care systems. The study also sought to develop an explanatory model for decision-making responses to febrile illnesses and skin conditions. METHODOLOGY: By using a non-experimental, cross-sectional study design and focused ethnographic approach, a sample of 200 Nasioi community members were interviewed by Nasioi-speaking research assistants. RESULTS: The study found that people in the sample group subscribe to both traditional and western medical paradigms. Western medical concepts have been assimilated but have not displaced traditional understanding of illness. There was congruence between beliefs about causes of illness, treatment-seeking responses to illness and stated or hypothetical preferences for traditional or western medicine. Data obtained in each of these domains reflect concepts of illness derived from both medical paradigms and demonstrate participants' confidence in the efficacy of both traditional and western medicine. CONCLUSIONS: It is proposed that a health system that incorporates traditional medicine may be better aligned with people's concepts of illness than the current system. Because it is more consistent with Nasioi concepts of illness, an incorporated health system may lead to more appropriate health service utilisation and, ultimately, to improvements in population health status. PMID- 19280445 TI - Moderators of and mechanisms underlying stereotype threat effects on older adults' memory performance. AB - Recent research has suggested that negative stereotypes about aging may have a detrimental influence on older adults' memory performance. This study sought to determine whether stereotype-based influences were moderated by age, education, and concerns about being stigmatized. Possible mechanisms underlying these influences on memory performance were also explored. The memory performance of adults aged 60 to 70 years and 71 to 82 years was examined under conditions designed to induce or eliminate stereotype threat. Threat was found to have a greater impact on performance in the young-old than in the old-old group, whereas the opposite was observed for the effects of stigma consciousness. In both cases, the effects were strongest for those with higher levels of education. Further analyses found little evidence in support of the mediating roles of affective responses or working memory. The only evidence of mediation was found with respect to recall predictions, suggesting a motivational basis of threat effects on performance. These findings highlight the specificity of stereotype threat effects in later adulthood as well as possible mechanisms underlying such effects. PMID- 19280447 TI - Audiovisual speech in older and younger adults: integrating a distorted visual signal with speech in noise. AB - Age-related changes to sensation and cognition may contribute to the differences found between younger and older adults using audiovisual speech. To evaluate this issue, younger and older adults repeated the final word of audio or audiovisual sentences in background noise. Although younger adults performed better than older adults, both age groups demonstrated the same level of visual enhancement in normal audiovisual conditions. However, when the visual speech was blurred, older adults had almost no audiovisual enhancement, whereas younger adults' enhancement was unaffected. These findings are discussed in terms of their implications for audiovisual integration over the course of the life span. PMID- 19280446 TI - Genetic and environmental mediation of the associations between self-rated health and cognitive abilities. AB - This study examined the relationship between self-rated health and cognitive abilities. First, the authors investigated whether common complex diseases mediate the associations between self-rated health and cognitive abilities. Slight evidence was found that associations between self-rated health and cognitive test scores were mediated by chronic diseases. Second, the authors explored the genetic and environmental mediation of these relationships using bivariate quantitative genetic analyses. In the younger age group (<67 years), associations between self-rated health and spatial reasoning and perceptual speed were mediated by both genetic and nonshared environmental factors. In the oldest age group (>or=67 years), associations between self-rated health and spatial reasoning, perceptual speed, and visual memory were entirely due to genetic factors. PMID- 19280448 TI - The effects of chemotherapy on Useful Field of View (UFOV) in younger and older breast cancer patients. AB - Chemotherapy is thought to cause cognitive deficits in some breast cancer patients, but the relative effects on older and younger breast cancer patients are unknown. The effects of chemotherapy on everyday cognitive tasks have not been examined. Thirty-eight female breast cancer survivors (3 to 45 months post chemotherapy) were compared to 55 age-matched control participants. Participants completed the Useful Field of View (UFOV), a computerized test of visual information processing that has been shown to decline with age, and which has been used to predict older adults' driving performance. Older chemotherapy patients performed more poorly than controls on the UFOV speed of processing, but not on the other two components. They also performed more poorly than younger chemotherapy patients. On the divided attention and selective attention components of the UFOV, older participants performed more poorly than younger participants, but there were no significant differences between chemotherapy patients and controls. These findings are explained in terms of brain changes thought to be caused by chemotherapy, which might have the most impact on older adults, already at risk for behavioral slowing. PMID- 19280449 TI - Older adults' responses to emotional stimuli: a cautionary note. AB - Thirty younger (mean age = 21 years) and 30 older (mean age = 73 years) adults were compared to determine if they had similar affective experiences to eight emotion films previously validated with young adults (Gross & Levenson, 1995). Participants rated their emotions, and heart rate was collected during two films of each emotion: amusement, anger, sadness, and fear. Older and younger adults were generally similar in their physiological and subjective responses to films, but with a few exceptions. Older adults reported more intense anger in response to one film and more intense nontarget negative emotions for both anger films compared with young adults. Some older adults also reported a negative response compared with young adults to one of the films designed to elicit amusement. Thus it is inappropriate to assume that emotional stimuli produce the same response across the adult life span. PMID- 19280450 TI - Factor structure and invariance of the Quality of Life in Alzheimer's Disease (QoL-AD) Scale. AB - The factor structure and factorial invariance of the Quality of Life in Alzheimer's Disease (QoL-AD) Scale was investigated in a sample of 653 nondemented, community-dwelling older adults, ages 57 to 95 years (M = 71.62, SD = 8.86), from the Seattle Longitudinal Study. The total sample was split into two random halves to explore and confirm the structure of the QoL-AD. Confirmatory factor analyses indicated better fit for a three-factor solution than one- or two factor solutions. Weak factorial invariance was found for the three-factor solution (Physical, Social, and Psychological Well-being) across age group and gender. These findings may help to establish a baseline quality of life before the onset of any noticeable AD symptoms. PMID- 19280451 TI - Life span adult faces: norms for age, familiarity, memorability, mood, and picture quality. AB - Because faces are frequently used as stimuli in cognitive aging behavioral and neuroimaging studies, there is a need for a normed set of pictures reflecting age and ethnic diversity. Minear and Park (2004, Behavior Research Methods, Instruments, & Computers, 36, 630-633) provided a large database of face pictures from average, ethnically diverse individuals across the adult life span. The current study evaluated 180 pictures from that database. One hundred and eight participants (mean age 23) rated each picture on specific attributes: perceived age, familiarity of appearance, mood, memorability, and image quality. Because the pictures did not differ in any of the measured attributes, the authors provide three balanced sets of face stimuli to researchers desiring normed face stimuli free from age confounds. PMID- 19280452 TI - Ethanol-mediated expression of connective tissue growth factor (CCN2) in mouse pancreatic stellate cells. AB - Activated pancreatic stellate cells (PSC) play a central role in the pathogenesis of pancreatic fibrosis, a common feature of chronic pancreatitis which is often caused by excessive alcohol consumption. In view of the central role of connective tissue growth factor (CCN2) in fibrosis, we investigated the mechanisms by which CCN2 is regulated in PSC following their exposure to ethanol or acetaldehyde. Primary cultures of PSC from Balb/c mice were treated with 0-50 mM ethanol or 0-200 microM acetaldehyde in the presence or absence of 4 methylpyrazole (4MP; an inhibitor of alcohol dehydrogenase), diallyl sulfide (DAS; an inhibitor of cytochrome P4502E1) or anti-oxidant catalase or vitamin D. CCN2 production, assessed by reverse-transcriptase polymerase chain reaction to measure CCN2 mRNA levels or by fluorescence activated cell sorting to assess CCN2 protein, was enhanced in a dose-dependent manner by ethanol or acetaldehyde. In the presence of 4MP, DAS, or the anti-oxidants vitamin D or catalase, there was a substantial decrease in the ability of ethanol to stimulate CCN2 mRNA expression and a concomitant decrease in CCN2-positive PSC. Accumulation of reactive oxygen species in PSC after exposure to ethanol was verified by loading the cells with dichlorofluorescin diacetate and showing that there was a stimulation of its oxidized fluorescent product, the latter of which was diminished in the presence of catalase or vitamin D. These results show the production of acetaldehyde and oxidant stress in mouse PSC are the cause of increased CCN2 mRNA and protein production after exposure of the cells to ethanol. The potential therapeutic effects of inhibitors of ethanol metabolism or anti-oxidants in alcoholic pancreatitis may arise in part through their ability to attenuate CCN2 production by PSC. PMID- 19280453 TI - Investigating the effects of cancer risk and efficacy perceptions on cancer prevention adherence and intentions. AB - This study examined the interaction between comparative cancer risk and efficacy perceptions on individuals' adherence for colon, prostate, and breast cancer screenings, intentions to get these screenings in the future, and intentions to adopt health lifestyle behaviors in the next year. A national probability sample of 2,226 adults ages 40 to 70 was surveyed. Overall, a positive interaction effect was found between comparative risk and efficacy on several outcomes. There were some methodological limitations worth noting, but the findings do have implications for health campaigns, particularly the need to increase efficacy beliefs about reducing cancer risks within the general population. PMID- 19280454 TI - Investigating the effect of nurse-team communication on nurse turnover: relationships among communication processes, identification, and intent to leave. AB - Enhanced team communication may strengthen nurses' attachment to their organizations and teams and improve nurse retention. This study examines the relationships among nurse-team communication, identification (organizational and team), and intent to leave. Hospital nurses (N = 201) completed surveys measuring 3 nurse-team communication processes: promoting team synergy, ensuring quality decisions, and individualizing communication. Hierarchical regression analyses revealed that promoting team synergy was a significant predictor of intent to leave, whereas ensuring quality decisions and individualizing communication did not account for significant additional variance in intent to leave. Separate analyses showed that the relationship between promoting team synergy and intent to leave was partially mediated by team identification or by organizational identification. Further analyses were conducted on the 7 communication practices for promoting team synergy. Mentoring emerged as the only significant predictor of intent to leave; however, its relationship to intent to leave was fully mediated by organizational identification or partially mediated by team identification. Pragmatic suggestions are offered to improve nurse identification and reduce turnover. PMID- 19280455 TI - Design and evaluation of a computer game to promote a healthy diet for young adults. AB - This article reports the development and evaluation of a computer game (RightWay Cafe) as a special medium to promote a healthy diet for young adults. Structural features of computer games, such as interactive tailoring, role playing, the element of fun, and narrative, were operationalized in the RightWay Cafe game to afford behavior rehearsal in a safe and entertaining way. Theories such as the health belief model, social cognitive theory, and theory of reasoned action guided the content design of the game to influence mediators of behavior change, including self-efficacy, perceived benefits, perceived barriers, and behavior change intention. A randomized controlled evaluation study with pretest, posttest, and follow-up design demonstrated that this game was effective in teaching nutrition and weight management knowledge and increasing people's self efficacy and perceived benefits of healthy eating, as well as their intention to be on a healthy diet. Limited long-term effects were also found: participants in the game-playing group had greater self-efficacy than participants in the control group after 1 month. This study validates the computer game-based approach to health promotion for young adults. Limitations and implications are also discussed. PMID- 19280456 TI - Humorous communication within the lives of older adults: the relationships among humor, coping efficacy, age, and life satisfaction. AB - This study explores humorous communication within the lives of older adults by testing the relationships among humor, coping efficacy, age, and life satisfaction. Results overall indicate these four variables function in communicatively complex ways for older adults. As predicted, humor-oriented individuals were more likely to use humor as a coping mechanism and reported greater coping efficacy. In a comparison of individuals who varied in age (under 50, 50-74, and 75 + ), differences in humor orientation scores emerged among the 3 groups. As predicted by Folkman and Lazarus's transactional theory of coping and emotion, the relationship between self-reported humor orientation and life satisfaction was mediated by coping efficacy. In attempting to determine which variables best predicted life satisfaction, coping efficacy and health status explained unique variance in life satisfaction scores. Study implications and limitations are addressed. PMID- 19280457 TI - A 15-year review of ABC, CBS, and NBC news coverage of organ donation: implications for organ donation campaigns. AB - This content analysis represents news coverage of organ donation from January 1990 to December 2005. Specifically, ABC, CBS, and NBC news broadcasts were examined to gain a greater understanding of organ donation coverage on TV. Overall this investigation revealed that organ donation received modest coverage (N = 1,507). Although the majority of coverage was positive, attention to the need for organs and the process of becoming a potential organ donor received modest exposure. In addition, non-living donor and living-donor donations received approximately equal coverage. Results are discussed with a focus on message design for practitioners and advocates of organ donation. PMID- 19280458 TI - Medical students' interviews with older adults: an examination of their performance. AB - During a geriatrics/gerontology curriculum designed to develop positive attitudes toward older adults, promote understanding about the psychosocial aspects of normal aging, and provide experience with intergenerational communication, students in a combined BA/MD degree program participated in learning activities that focused on interacting with and interviewing older adults. This article describes medical students' performance during an intergenerational interview, examines differences between students' self-assessments and evaluators' ratings of students' performance, and presents statistical relationships among communication apprehension, situation anxiety, competence, and performance. Data analysis indicates that the majority of students demonstrated competent intergenerational interviewing skills during their standardized communication assessment. The discussion elaborates on the study findings, acknowledges limitations, and addresses practical implications of the study. PMID- 19280459 TI - Religiosity, anxiety, and discussions about organ donation: understanding a complex system of associations. AB - An increasingly large research base on religiosity has shown it to have a buffering effect on anxiety. In a separate vein, scholars interested in organ donation have suggested that both religiosity and anxiety play roles in individuals' willingness to seek information concerning their decisions about organ donations with their family-an event that greatly increases donation rates. This investigation presents 2 studies that examine the associations between religiosity and anxiety (variously measured), on the one hand, and anxiety and individual's information seeking behaviors with family members about organ donation on the other. The first study offers national samples and relies on self reports, whereas the second study is one of the few organ donation studies to provide observer ratings of interaction between family members on the issue. Results suggest a more complicated role of religiosity with regard to anxiety than previously believed and show a consistent and robust association between anxiety and communication behaviors regarding organ donation. Implications for campaigns are discussed. PMID- 19280460 TI - Online physical activity information: will typical users find quality information? AB - This study evaluated physical activity Web sites to determine quality, accuracy, and consistency with principles of the extended parallel process model (EPPM). Three keyword searches were conducted using 4 search engines to find a sample of N = 41 Web sites. Three raters evaluated the Web sites using the JAMA benchmarks to assess quality and American College of Sports Medicine and Centers for Disease Control and Prevention recommendations for physical activity to determine accuracy, as well as checking for inclusion of EPPM variables. Data were analyzed using descriptive statistics and analysis of variance with least squares means. Only 22% of the sites were high quality, none were highly accurate, and most were consistent with the EPPM. Quality ratings were weakly associated with accuracy. Educational and .net sites were rated significantly higher in quality and accuracy, and government sites were most consistent with the EPPM. Quality Web sites were more often found by using Yahoo and Google. "Exercise" yielded more accurate results, whereas "physical activity" and "fitness" produced more Web sites consistent with the EPPM. It is encouraging that most sites incorporated EPPM concepts; however, quality and accuracy were poor, leaving physical activity information seekers at risk for disease and injury. PMID- 19280461 TI - The importance of being authentic: persuasion, narration, and dialogue in health communication and education. AB - A topic gaining attention in the field of education has been that of "authenticity"-the creation of learning environments that provide learners with richer and more realistic contexts in which to apply knowledge and practice skills. The subject of authenticity has yet to attract much attention in the field of public health and specifically in health communication and education, although these fields have long experience with the problem of getting audiences to translate knowledge into action. This article reviews the rationale for an interest in authenticity as it relates to health communication and education and notes that "authenticity" does not inhere in information but is an appraisal made by a member of the public who is persuaded to view the information as especially relevant to his or her health behavior and consonant with his or her prior experiences. This article argues that a public health communicator or educator can encourage such appraisals by using narrative formats that provide rich contextualization. But contextualizing behavior change information in the form of stories is not enough; there is a dialogic dimension to persuasion that aids in the process of authentication. Creating opportunities for dialogue between behavior change narratives and their audiences has its own challenges, but nonetheless deserves to be a priority in public health. PMID- 19280463 TI - The role of apoptosis in radiation oncology. AB - PURPOSE: Apoptosis, as a mode of cell death in irradiated cell populations, has been the subject of literarily hundreds if not thousands of published reports over the past few years. However, in spite of the large body of knowledge related to this subject, the role of apoptosis in determining tumor response to radiotherapy has been and remains poorly understood and controversial. Indeed, some previous reviews have suggested that apoptosis may not be important in this context. The purpose of the present review is to provide some examples of recently reported laboratory investigations that indicate that there is a reasonable expectation that the radiation-induced apoptosis observed has contributed to the tumor response. CONCLUSIONS: We review reports in four areas of research: Molecularly targeted agents, in vivo imaging, Bcl-2 and cancer stem cells. Examples are provided in each of these areas that we believe justify a reassessment of the role that apoptosis plays in radiation oncology. PMID- 19280464 TI - Protection of mice against X-ray injuries by the post-irradiation administration of guanosine and inosine. AB - PURPOSE: To examine the radioprotective action of guanosine (Guo) and inosine (Ino) administered to mice after irradiation with X-rays. MATERIALS AND METHODS: Survival of mice exposed to lethal and sublethal doses of X-rays was studied. Peripheral blood cells were counted using a light microscope. The damage to bone marrow cells was assessed by micronucleus (MN) test. Damage and repair of DNA in blood leukocytes were estimated using the comet assay. RESULTS: Mice injected intraperitoneally (i.p.) with Guo or Ino ( approximately 30 microg g(-1), i.e., approximately 0.6 mg per 20-g mouse) 15 min after acute whole-body irradiation with 7 Gy recovered from X-ray injury. On the 30th day after irradiation, 50 and 40% of mice injected with Guo and Ino, respectively, remained alive. The dose reduction factor (DRF) was 1.23 for Guo and 1.15 for Ino. The protective effect gradually decreased as the time interval between the irradiation and injection was increased to 3, 5, 8 h. Guo and Ino facilitated the restoration of peripheral blood cell counts. These compounds protected bone marrow cells from damage and normalized erythropoiesis. Guo and Ino contributed to a more rapid and complete repair of DNA in mouse leukocytes irradiated both in vitro and in vivo. CONCLUSION: Guo and Ino introduced shortly after irradiation reduce leukopenia and thrombocytopenia and offer promise as therapeutic agents for treatment of radiation injuries. PMID- 19280465 TI - Radiobiological evaluation and correlation with the local effect model (LEM) of carbon ion radiation therapy and temozolomide in glioblastoma cell lines. AB - PURPOSE: To investigate the cytotoxic effect of high linear-energy transfer (LET) carbon irradiation on glioblastoma cells lines in combination with temozolomide (TMZ). METHODS AND MATERIALS: The cell lines U87-MG expressing wild-type p53 and LN229 expressing both mutant and wild-type p53 were irradiated with monoenergetic carbon ion beams (LET 172 keV/microm) or an extended Bragg peak (LET 103 keV/microm) after treatment with 10 microM or 20 microM TMZ. Cytotoxicity was measured by a clonogenic survival assay, and cell growth as well as cell cycle progression, were examined. RESULTS: The p53 mutant was more sensitive to X-ray irradiation than the p53 wild type cell line, which was also expressed in a shorter G2 block. High LET carbon ions show an increased biological effectiveness in both cell lines, which is consistent with the predictive calculations by the Local Effect Model (LEM) introduced by Scholz et al. The cell line LN229 was more sensitive to TMZ treatment than the U87MG cell line expressing wild-type p53 only. The combination of TMZ and irradiation showed an additive effect in both cell lines. CONCLUSION: High LET carbon ion irradiation is significantly more effective for glioblastoma cell lines compared to photon irradiation. An additional treatment with TMZ may offer a great chance especially for several tumor types. PMID- 19280466 TI - Radiation-induced matrix production of lung fibroblasts is regulated by interleukin-8. AB - PURPOSE: Lung fibrosis can be caused by radiation therapy during cancer treatment and therefore can be the limiting factor of the treatment. The factors that cause the actual fibrosis and the interaction between different cell types were investigated. MATERIALS AND METHODS: Epithelial lung cells and fibroblasts were irradiated and different cytokines were measured in the supernatant. Also effects of radiation on the matrix production of fibroblasts were investigated. RESULTS: Irradiation of isolated lung fibroblasts did not cause increased extracellular matrix production; however, the co-culturing of fibroblasts and irradiated lung epithelial cells or the treatment of fibroblasts with supernatants of irradiated epithelial cells did result in an increase. We were able to show that increased interleukin-8 (IL-8) levels led to increased matrix production. CONCLUSIONS: IL-8 is not only a proinflammatory cytokine but it also stimulates collagen synthesis and matrix production and therefore could be a possible drug target in preventing radiation damage during cancer therapy. PMID- 19280467 TI - Assessment of genetic damage in peripheral blood of human volunteers exposed (whole-body) to a 200 muT, 60 Hz magnetic field. AB - AIM: To investigate the extent of damage in nucleated cells in peripheral blood of healthy human volunteers exposed to a whole-body 60 Hz, 200 microT magnetic field. MATERIALS AND METHODS: In this study, 10 male and 10 female healthy human volunteers received a 4 h whole-body exposure to a 200 microT, 60 Hz magnetic field. In addition, five males and five females were treated in a similar fashion, but were exposed to sham conditions. For each subject, a blood sample was obtained prior to the exposure period and aliquots were used as negative- (pre-exposure) and positive- [1.5 Gray (Gy) (60)Cobalt ((60)Co) gamma irradiation] controls. At the end of the 4 h exposure period, a second blood sample was obtained. The extent of DNA damage was assessed in peripheral human blood leukocytes from all samples using the alkaline comet assay. To detect possible clastogenic effects, the incidence of micronuclei was assessed in phytohemagglutinin (PHA)-stimulated lymphocytes using the cytokinesis-block micronucleus assay. RESULTS: There was no evidence of either increased DNA damage, as indicated by the alkaline comet assay, or increased incidence of micronuclei (MN) in the magnetic field exposed group. However, an in vitro exposure of 1.5 Gy gamma-irradiation caused a significant increase in both DNA damage and MN induction. CONCLUSIONS: This study found no evidence that an acute, whole-body exposure to a 200 microT, 60 Hz magnetic field for 4 hours could cause DNA damage in human blood. PMID- 19280468 TI - Protective effects of insulin-like growth factor-I on the decrease in myogenic differentiation by ionizing radiation. AB - PURPOSE: The aim of the work is to evaluate the effects of insulin-like growth factor-1 (IGF-1) on the decrease in myotube formation induced by ionizing radiation. MATERIALS AND METHODS: We induced C2C12 cells to a myogenic linage following X-ray irradiation at 2 and 4 Gy. Myogenic differentiation was estimated using immnocytochemical staining with anti-myosin antibody, and the anti-myosin antibody positive areas, the total number of nuclei, the number of nuclei included in multinucleated myotubes per field, and the myotube formation ratio were analyzed. RESULTS: In the myogenic differentiation in the presence of IGF-1, the decrease in anti-myosin antibody positive areas, the nuclei included in myotubes, and the myotube formation ratio induced by X-ray irradiation at 2 Gy was restored to control levels. CONCLUSIONS: The addition of IGF-1 protected against the decrease myotube formation induced by X-ray irradiation at 2 Gy. Since X-ray irradiation at 2 Gy is usually used for multi-fractionated irradiation in radiotherapy, our findings suggest that IGF-1 could be useful to protect against impairment of muscle repair induced by therapeutic dose radiation. PMID- 19280469 TI - Evaluation of radioadaptive response induced in CHO-K1 cells in a non-traditional model. AB - PURPOSE: The present study was designed to evaluate the effects of sequential exposures to low doses of gamma-radiation that induce a radioadaptive response to a later high-dose of radiation in CHO-K1 cells. MATERIALS AND METHODS: Cells were cultured in four dilution cycles and grown to confluency. Radiation treatment was performed once per cycle with 0.1 Gy gamma-rays. After the last radiation period (chronic radiation) the culture was irradiated with a higher dose (1 Gy). Each cell culture was immediately divided into two fractions: one of them was used to carry out the comet assay and the other for the structural chromosome aberration test. In the first fraction, genotoxic damage was evaluated by degree of damage in 300 cells per experimental point. The second assay was performed with 400 cells per treatment. The statistical analysis was carried out using the chi(2) test. RESULTS: Results from these assays demonstrated a genotoxic effect for both the adaptive and acute treatments (p < 0.001). The comet assay showed a significant increase in damage for the combined treatment when compared with 1 Gy treatment (p < 0.001). The frequency of chromosomal aberrations (CA) was lower for the combined treatment than for that using the highest radiation dose. CONCLUSIONS: These results suggest the possible induction of a radioadaptive response after the sequential exposure to very low doses of radiation. The finding of decreased cytogenetic damage after one cell cycle and not immediately after radiation could indicate the eventual potentiation of repair mechanisms. PMID- 19280470 TI - 1-Methylxanthine enhances the radiosensitivity of tumor cells. AB - PURPOSE: To determine the efficacy of a caffeine derivative 1-methylxanthine (1 MTX) in increasing radiosensitivity of cancer cells and elucidate the underlying mechanisms in vitro. MATERIALS AND METHODS: RKO human colorectal cancer cells carrying wild type protein 53 kDa (p53) were incubated with 3 mM 1-MTX for 30 min, exposed to 4 Gy ionizing radiation, and further incubated with 1-MTX for three days. The clonogenic cell death was determined, and the cell cycle distribution and apoptosis were studied with flow cytometry at different times after irradiation. The DNA double strand break (DNA DSB) was examined using phosphorylated Histone2A (gamma-H2AX) foci formation, and the expression/activity of checkpoint 2 kinase (Chk2), cell division cycle 25 (Cdc25) phosphatase and cyclin B1/Cdc2 kinase were also investigated using western blotting and in vitro kinase assays. RESULTS: The treatment with 3 mM 1-MTX increased the radiation induced clonogenic and apoptotic cell death. The radiation-induced phosphorylation of Chk2 and Cdc25c and the radiation-induced increase in the cyclin B1/Cdc2 kinas activity were little affected by 1-MTX. The radiation induced G2/M arrest was only slightly shortened and the expression of radiation induced gamma-H2AX was markedly prolonged by 1-MTX. CONCLUSIONS: 1-MTX significantly increased the radiosensitivity of RKO human colorectal cancer cells carrying wild type p53 mainly by inhibiting the repair of radiation-induced DNA DSB without causing significant alteration in radiation-induced G2/M arrest. Such a radiosensitization occurred at 1-MTX concentrations almost non-toxic to the target tumor cells. PMID- 19280471 TI - Review of advances in the thin layer chromatography of pesticides: 2006-2008. AB - Applications of thin layer chromatography and high performance thin layer chromatography for the separation, detection, qualitative and quantitative determination, and preparatire isolation of pesticides and their metabolites are reviewed for the period from July 1, 2006 to November 1, 2008. Analyses are described for a variety of sample types and pesticide classes. In addition to references on residue and formulation analysis, studies such as pesticide structure relationships, identification and characterization of plant pesticides, metabolism, degradation, mobility, and lipophilicity are covered. Future prospects for pesticide thin layer chromatography are also considered. PMID- 19280472 TI - Evaluation of a multiresidue method for pesticides in cereals using supercritical fluid extraction and gas chromatographic detection. AB - Supercritical fluid extraction (SFE) was evaluated to be applied for residue analysis of 22 gas chromatography/electron capture detector-nitrogen phosphorus detector (GC/ECD-NPD) amenable pesticides in rice, wild rice and wheat. Samples were extracted with supercritical carbon dioxide at 200 atm pressure and 50 degrees C temperature, using methanol as a static modifier. Mean recoveries obtained with the proposed SFE method at two spiking levels with four replicates per level are compared with those obtained with an ethyl acetate-based solvent extraction/gel permeation chromatography (GPC) clean up method. Both methods gave consistent high recoveries for almost all the pesticides from all the commodities with overall mean recoveries higher than 70% with relative standard deviations lower than 20%. Remarkable exceptions were captafol and dimethoate, for which low and/or non-reproducible recoveries were obtained with the SFE method. Residue levels determined with both methods in nine different incurred samples of wheat, containing some of the studied pesticides, were very similar, but, in all cases, slightly higher levels were determined with the SFE method. PMID- 19280473 TI - Organo-mineral interactions mask the true sorption potential of biochars in soils. AB - The sorption of carbaryl (1-naphthyl methyl carbamate) and ethion [O,O,O',O' tetraethyl S,S'-methylene bis(phosphorodithioate)] was studied in whole soils as well as after treatment of soil with 2% hydrofluoric acid (HF) to remove paramagnetic materials and to oxidize most forms of labile carbon by photo oxidation with high energy (UV) on < 53 microm fractions. The sorption coefficient (K(d)) values for carbaryl and ethion in soils did not follow the order of their organic carbon (OC) content, and specially their char content However, the K(oc) values in < 53 microm fractions after hydrofluoric acid/photo oxidation with high energy (hydrofluoric acid/ultraviolet; HF/UV) treatment were found to be much higher than those in bulk untreated soils. The effect of organic matter chemistry was determined by correlating K(oc) values of contaminants in bulk soils or 53 microm fractions against sample aromaticity. A poor correlation of K(oc) in bulk soil and aromatic C values of both carbaryl and ethion was observed. However, the correlation between the K(oc) and the aromatic fraction of C after the HF/UV treatment improved significantly, reflecting the contribution of char fraction of carbon in soils towards sorption of pesticides. The increase in sorption after HF/UV treatment suggested that the sorption potential of biochars, which are expected to contribute significantly to contaminant sorption due to their high surface area, can remain masked by the organo-mineral interactions of char in whole soils. This has implications for the modification of surfaces of the freshly applied biochars in soils due to organo-mineral interactions. PMID- 19280474 TI - Removal of fenhexamid and pyrimethanil from aqueous solutions by clays and organoclays. AB - The ability of a sodium montmorillonite (CLONa) and two commercial available organoclays having interlayer organic cations possessing different functional groups (CLO20A and CLO30B) was investigated for adsorbing two pesticides namely fenexamid (FEX) and pyrimethanyl (PMT). The two organoclays displayed a higher affinity with the pesticides than the unmodified clay, but the improvement in adsorption capacity varied according to the characteristics of the pesticide and the interlayer organic cation. FEX was adsorbed to a greater extent than PMT by both organoclays, which may be due to the higher hydrophobicity of FEX thereby indicating considerable hydrophobic interaction between the adsorbent/adsorbate systems. Our findings may find application in the removal of water-soluble pesticides from aquifers. PMID- 19280475 TI - Effects of thiobencarb in combinations with molinate and chlorpyrifos on selected soil microbial processes. AB - The impact of pesticides, namely thiobencarb (TBC), molinate (MOL) and chlorpyrifos (CPF), on soil microbial processes was studied in two Australian soils. Substrate induced respiration (SIR), substrate induced nitrification (SIN) and phosphatases and chitinase enzymatic activities were assessed during a 30-day microcosm study. The pesticides were applied to soils at recommended rates either alone, or as binary mixtures with TBC. Soil samples were sampled at 5, 15 and 30 days after pesticide treatments. Substrate induced respiration was only transiently affected by pesticides in both soils. In contrast, the process of indigenous nitrification was affected by the presence of pesticides in both soils, especially when the pesticides were applied as binary mixtures. Substrate induced nitrification increased with pesticides in the Griffith soil (except with MOL+TBC after 5 days) whereas SIN values were non-significantly different to the control on the Coleambally soil. The binary mixtures of pesticides with TBC resulted in a decrease in SIN in both soils, but the effects disappeared within 30 days. The enzymatic activities were not consistently affected by pesticides, and varied with the soil and pesticides studied. This study showed that, when applied at recommended application rates, TBC, MOL, and CPF (individually or as binary mixtures), had little or only transitory effects on the functional endpoints studied. However, further investigations are needed to assess the effect on microbial densities and community structure despite the low disturbance to the functions noted in this work. PMID- 19280476 TI - Solubility and adsorption behaviors of chlorpyriphos-methyl in the presence of surfactants. AB - In the present work changes in the adsorption of the pesticide chlorpyrifos methyl (CLP-m) on soil colloids induced by application of surfactants were determined using a batch equilibrium method. The surfactants used were sodium dodecyl sulphate (SDS), Tween 20, and dihexadecyldimethylammonium bromide (DHAB). The adsorption isotherms of CLP-m in aqueous medium and in surfactant solutions at concentration equal to the critical micelle concentration (CMC) fitted the Freunlich adsorption equation generally with R(2) values greater than 0.96. While the addition of SDS and DHAB decreased the pesticide adsorption, the addition of Tween 20 increased the pesticide adsorption. The increases or decreases in the adsorption in the experiment revealed that the behavior of CLP-m in soil water systems mainly depends on the type of surfactant. Moreover water solubility of CLP-m changes by the three surfactants below and above their CMC were studied. While the solubility of CLP-m was enhanced by SDS both below and above the CMC, the solubility of the pesticide was enhanced by DHAB only above the CMC. Tween 20 did not influence the solubility of CLP-m. PMID- 19280477 TI - Degradation behavior of sulfadiazine in soils under different conditions. AB - This study investigated the degradation of sulfadiazine in three soils and also determined its sorption and hydrolysis behaviors as well. At the spike concentration of 10 mg/kg, the half-lives for sulfadiazine in the aerobic nonsterile soils ranged from 12 days to 18 days. Sulfadiazine was more persistent in the anoxic soils with the half-lives ranging between 57 days and 237 days and soil microorganisms played little role in the dissipation process under anoxic conditions. The decline in sulfadiazine concentrations was also observed in the sterile soils under aerobic conditions. Hydrolysis could not explain this phenomena as hydrolysis of sulfadiazine was pH dependent. Sulfadiazine only hydrolyzed to a very limited degree at acidic pH. Increased sorption was observed for sulfadiazine in soil 1 (pH 4.3) when the contact time increased to 14 days, but no significant increase in sorption was found for soil 2 (pH 7.2) and soil 3 (pH 8.5). PMID- 19280478 TI - Transport and deposition of organochlorine pesticides from farmland to estuary under tropical regime and their potential risk to aquatic biota. AB - In this study, residues of organochlorine pesticides (OCPs) in water, suspended particulate matter (SPM), sediment and oysters from the Chanthaburi estuary were monitored closely in the wet and dry seasons for determining potential environmental risks. In water samples, out of twenty OCPs measured, endosulfan and hexachlorocyclohexanes (HCHs) were the two predominant compounds ranging and concentrations between 0.6-13 ng/L and 1-12 ng/L, respectively. Both compounds were found in higher concentrations during the wet season. The two insecticides were significantly correlated with total organic carbon (TOC) in the water with r values of 0.571 (P < 0.05) and 0.440 (P < 0.1), for endosulfan and HCHs respectively suggesting that these chemicals are strongly bound and concentrated by water-soluble organic carbon. As expected, high concentrations of endosulfan and HCHs were associated with SPM and in the surface sediment (0-5 cm) with a concentration ranging from 25-1,219 microg/kg dry wt and 19-110 microg/kg dry wt for SPM, 4-70 microg/kg dry wt and 0.5-50 microg/kg dry wt for surface sediment, respectively. Sediment quality of the Chanthaburi estuary was also assessed in terms of risk to aquatic biota using concentration found in the sediment compared to published allowable threshold levels (U.S. EPA and New York guidelines). Results suggested that most of endosulfan and HCHs levels in sediment samples exceeded the threshold values. Thus potential and environmental risks may be anticipated from levels found in sediment. Oyster from selected farms cultured at the study area appeared to take up both endosulfan and HCHs in proportion to level presented in the water and SPM. The range of endosulfan and HCHs observed in oyster tissue were from 4-46 microg/kg wet wt, and from non-detectable to 8 microg/kg wet wt. respectively. Human risk from oyster consumption was considered by comparing the value measured with allowable threshold level of concerned chemical. The levels of endofulfan and HCHs measured in oyster showed that oysters from this estuary area were safe for consumption. PMID- 19280479 TI - Removal of mixed pesticides from drinking water system by photodegradation using suspended and immobilized TiO2. AB - Lindane (1alpha, 2alpha, 3beta, 4alpha, 5alpha, 6beta-hexachloro cyclohexane), methyl parathion (O,O-dimethyl-O-4-nitrophenyl phosphorothioate) and dichlorvos (2,2-dichlorovinyl-O-O-dimethyl phosphate) are removed from water individually and as a mixture by photo degradation using suspended and immobilized forms of TiO(2) (Degussa P-25). Studies were conducted to optimize the coating thickness of immobilized photo catalyst. The rate of degradation of pesticides was compared in both suspended and immobilized TiO(2) systems. Degradation studies of mixed pesticides were carried out with low concentrations (1.0 and 2.5 mg/L) of pesticides. Only three intermediate byproducts such as methyl paraoxon, O,O,O trimethyl phosphonic thionate and p-nitrophenol were observed during the methyl parathion degradation in suspended, immobilized TiO(2) systems and mixed pesticides degradation studies. At the end of the reaction methyl parathion and its by-products were completely degraded. During lindane degradation hexachloro cyclohexane, pentachloro cyclohexane, hexachloro benzene, 1-hydroxy 2,3,4,5,6 chlorocyclohexane, 1-hydroxy 2,3,4,5,6-chlorobenzene, pentachloro cyclopentadiene, 1,2,3,4,5-hydroxy cyclopentene and 1,2,3-hydroxy cyclobutane were identified in suspended and immobilized TiO(2) systems, whereas only hexachloro cyclohexane, pentachloro cyclohexane, hexachloro benzene and pentachloro cyclopentadiene were observed during mixed pesticides degradation. No intermediate by-product was observed during the photo degradation of dichlorvos. Langmuir-Hinshelwood pseudo first order kinetic equation showed that there was not much change in the rates of degradation in both suspended and immobilized TiO(2) systems irrespective of the pesticide. During mixed pesticides degradation, the degradation pattern was not similar to that of single pesticide. PMID- 19280480 TI - Pesticide exposure and blood endosulfan levels after first season spray amongst farm workers in the Western Cape, South Africa. AB - The study investigated serum endosulfan changes resulting from occupational exposure to the pesticide on farms. Eight applicators and 17 non-applicators were tested (serum endosulfan, anthropometry, short exposure questionnaire) before and after the first day of seasonal spraying. Task-based job exposure matrix (JEM) estimates were calculated. Mean baseline serum endosulfan (530 +/- 0.05 microg/L) was high. Increases in post-spraying endosulfan levels (IPSE) were higher in applicators (mean = 60 +/- 90 microg/L) than in non-applicators (mean = 3.5 x 10( 6)+/- 90.0 microg/L) adjusting for age (beta = 54.0, p = 0.162, R(2) = 0.22). There was a weak positive relationship between IPSE and JEM estimates. IPSE occurred in applicators and non-applicators and were higher in applicators. The validity of the JEM weightings and characterization of other routes of pesticide exposure require further investigation. PMID- 19280481 TI - Quantitative structure-activity relationship for the depuration rate constants of polychlorinated biphenyls in the freshwater mussel, Elliptio complanata. AB - Partial least squares (PLS) regression together with 16 theoretical molecular structural descriptors was successfully used to develop a quantitative structure activity relationship (QSAR) model on the depuration rate constants (k(d)) of polychlorinated biphenyls (PCBs) in freshwater mussel Elliptio complanata. The cross validated Q(2)(cum) and the correlation coefficient R for the model is 0.871 and 0.941, respectively. The achievement of satisfactory Q(2)(cum) and R values indicate significant high robustness and good predictive ability of the model. The model was tested as acceptable for prediction of logk(d) (the log of the depuration rate constants) by validation set. The significant descriptors governing logk(d) include polarizability (alpha), core core repulsion energy (CCR), molecular weight (M(w)), total energy (TE), and electronic energy (EE). The key descriptors in the model reflect that van der Waals interactions play a dominant role in the depuration of PCBs. The depuration of PCBs in Elliptio complanata may be mainly attributed to the biota-water phase partitioning processes. PMID- 19280482 TI - The effects of soil and Trifolium repens (white clover) on the fate of estrogen. AB - In this study, we investigated the behavior of estrogens in the rhizosphere of white clover (Trifolium repens, clover hereafter) with two different pot tests, using soil and agar as growth media. In a pot test using agar spiked with estrogen, the estrogen concentration in the agar with clover decreased to non detectable levels within one month, while in the agar without clover, 60% of initially added estrogen remained after one month. The half-lives of estrone (E1) and 17beta -estradiol (E2) in the agar with clover were 2.4-3.8 and 13.2 d, respectively. The dissipation of E1 followed first-order rate law, while that of E2 fitted a zero-order reaction, indicating that they had different mechanisms of dissipation from agar. In the soil pot test, the behavior of E1 and E2 was not influenced by clover. An initial rapid decrease in the amount of estrogen extracted by methanol/acetic acid was followed by persistence for 1-3 months, regardless of presence of clover. Moreover, in three weeks E1 and E2 were only partly degraded by microbes extracted from the soil used in the pot test. In this study, abiotic degradation of estrogens and sorption of estrogen to soil, rather than the effects of soil microbes and clover, contributed to the initial rapid dissipation of estrogens in the soil. However, the results of the agar pot test suggested that vegetation such as clover may significantly contribute to removal of estrogens when estrogens in aqueous phase are discharged with surface runoff and preferential flow after heavy rain in agricultural fields, or when present in soils with low estrogen sorptivity. PMID- 19280483 TI - Application of indoxacarb for managing shoot and fruit borer of eggplant (Solanum melongena L.) and its decontamination. AB - Indoxacarb was applied at 75 and 150 g a.i. ha(-1) for two years to an eggplant (Solanum melongena L.) crop grown in the field plots in order to evaluate its efficacy for management of the lepidopteron pest, shoot and fruit borer. The residues of the insecticide were quantified by high-pressure liquid chromatography (HPLC). The mean initial deposits of indoxacarb on eggplant fruits were found to be 2.60-2.634 mg kg(-1) and 3.64-3.68 mg kg(-1) from the two rates of applications, respectively. They declined with time and reached to non detectable (< 0.02 mg kg(-1)) after 15-20 d. Residues dissipated with a half-life of 3.0-3.8 d from both first and second-year application. A 3 d waiting period for harvest of fruits after insecticide application and processing resulted in the residue levels that were below the Codex maximum residue limit (MRL) of 0.5 mg kg(-1) thereby achieving a maximum safety and minimum risk to consumers. The best combination of chemicals for decontamination of indoxacarb was found to be by washing with a mixture of alkali and potassium permanganate (KMnO(4)) thereby resulting in the removal of 67.5% and 59.2 % residues for 5 and 10 microg g(-1) spiking doses, respectively. Major products formed on reaction of indoxacarb with alkali were identified by electron spray ionization chromatography/mass spectrometry (ESI/MS). The per cent reduction on the weight and number basis of treated eggplant plots were compared to those observed in control plots to demonstrate the effectiveness of indoxacarb treatment on shoot and fruit borer population. PMID- 19280484 TI - The use of naturally generated volatile fatty acids for herbicide removal via denitrification. AB - This research focuses on the removal of 2, 4-D via denitrification, with a particular emphasis on the effect of adding naturally generated volatile fatty acids (VFAs) as a carbon source. These VFAs had been produced from an acid-phase anaerobic digester (mean VFA concentration of 3153 +/- 801 mg/L [as acetic acid]). The first step involved developing 2, 4-D degrading bacteria in a sequencing batch reactor (SBR) fed with both sewage and 2, 4-D (30-100 mg/L). Subsequent denitrification batch tests demonstrated that the specific denitrification rate increased from 0.0119 +/- 0.0039 to 0.0192 +/- 0.0079 g NO(3)-N/g volatile suspended solids (VSS) per day, when using 2, 4-D alone versus 2, 4-D plus natural VFAs from the digester as a carbon source. Similarly, the specific 2, 4-D consumption rate increased from 0.0016 +/- 0.0009 to 0.0055 +/- 0.0021 g 2,4-D/g VSS per day, when using 2, 4-D alone as compared to using 2, 4-D plus natural VFAs. Finally, a parallel increase in the percent 2, 4-D removal was observed, rising from 28.33 +/- 11.88 using 2, 4-D alone to 54.17 +/- 21.89 using 2, 4-D plus natural VFAs. PMID- 19280485 TI - Screening Brassica species for glucosinolate content. AB - Glucosinolates (GSLs), a group of compounds found in Brassica plants, are toxic to some soil-borne plant pathogens because of the toxicity of their hydrolysis products, isothiocyanates. Other phytochemicals found in Brassica plants, such as phenols and ascorbic acid, may compliment the activity of GSLs. A survey of Brassica accessions from the national germplasm repository was conducted to identify potential cover crops that could be soil-incorporated for use as biofumigants. Ten Brassica accessions that demonstrated relative cold tolerance, rapid maturity, and superior biomass production were selected. The selected accessions were grown under three climatic conditions (fall greenhouse, winter high tunnel, and spring field) to investigate whether growing conditions affect their GSL, phenol, and ascorbic acid content. The selected accessions included seven accessions of Brassica juncea (Indian mustard), one of Brassica napus (oil seed rape), one of Brassica campestris (field mustard), and one of Eruca sativa (arugula). Separation of GSLs from the selected Brassica accessions was achieved using ion-exchange sephadex in disposable pipette tips. Quantification of total GSLs was based on inactivation of the endogenous thioglucosidase and liberation of the glucose moiety from the GSL molecule by addition of standardized thioglucosidase (myrosinase) and colorimetry. GSL concentration of greenhouse, high tunnel, and field-grown shoots (leaves and stems) averaged 24, 40 and 76 micromoles g(-1) fresh weight, respectively. Accessions of B. juncea generally had the highest GSL content. A comparison of accessions revealed that Ames 8887 of B. juncea contained the greatest GSL concentration, but had the lowest biomass yield and ascorbic acid concentration, in part because phytochemical concentration tended to be negatively correlated with biomass yield. More promising was B. juncea accession 'Pacific Gold' which coupled high biomass yield with above-average GSL production, but had low phenol and ascorbic acid concentration. We concluded that environmental stress on growing plants can increase the concentration of GSLs, ascorbic acid, and total phenols in Brassica shoots, but does not increase yields of these phytochemicals per unit area. PMID- 19280486 TI - Ammonia volatilization loss from surface applied livestock manure. AB - Ammonia (NH(3)) emission from livestock manures used in agriculture reduces N uptake by crops and negatively impacts air quality. This laboratory study was conducted to evaluate NH(3)emission from different livestock manures applied to two soils: Candler fins sand (CFS; light-textured soil, pH 6.8 and field capacity soil water content of 70 g kg(-1)) from Lake Alfred, Florida and Ogeechee loamy sand (OLS; medium-textured soil, pH 5.2 and field capacity soil water content of 140 g kg(-1)) from Savannah, Georgia. Poultry litter (PL) collected from a poultry farm near Douglas, Georgia, and fresh solid separate of swine manure (SM) collected from a farm near Clinton, North Carolina were used. Each of the soil was weighed in 100 g sub samples and amended with either PL or SM at rates equivalent to either 0, 2.24, 5.60, 11.20, or 22.40 Mg ha(-1) in 1L Mason jars and incubated in the laboratory at field capacity soil water content for 19 days to monitor NH(3) volatilization. Results indicated a greater NH(3) loss from soils amended with SM compared to that with PL. The cumulative NH(3)volatilization loss over 19 days ranged from 4 to 27% and 14 to 32% of total N applied as PL and SM, respectively. Volatilization of NH(3) was greater from light-textured CFS than that from medium-textured OLS. Volatilization loss increased with increasing rates of manure application. Ammonia volatilization was lower at night time than that during the day time. Differences in major factors such as soil water content, temperature, soil type and live stock manure type influenced the diurnal variation in volatilization loss of NH(3) from soils. A significant portion (> 50%) of cumulative NH(3) emission over 19 d occurred during the first 5-7 d following the application of livestock manures. Results of this study demonstrate that application of low rates of livestock manure (< or = 5.60 Mg ha(-1)) is recommended to minimize NH(3) emissions. PMID- 19280487 TI - Psychosomatic approaches to obstetrics, gynaecology and andrology--a review. AB - This review aims to clarify the scope and clinical importance of psychosomatic approaches to obstetrics, gynaecology and andrology. This gradually expanding sub specialty covers a wide domain of complex disease conditions that can be managed more effectively if the various biological, psychological and social aspects are recognised at the start and concurrent treatment initiated. The current need to practise biopsychosocial management of disease conditions is highlighted along with a description of what this would involve. The nine-field psychosomatic approach, which can be applied to everyday clinical encounters, has been illustrated. Clinical applications of the psychosomatic approach are discussed for various conditions including chronic pelvic pain, eating disorders, tokophobia, post-traumatic stress disorder, depression, menstrual disorders, infertility, bereavement and testicular cancer. Cultural considerations and the need for further research are also briefly discussed. PMID- 19280488 TI - The effectiveness of discontinuing iron-containing prenatal multivitamins on reducing the severity of nausea and vomiting of pregnancy. AB - Nausea and vomiting of pregnancy (NVP) is experienced by the majority of pregnant women, and can negatively affect a women's quality of life. It has been suggested in observational studies that iron-containing prenatal multivitamins may increase the severity of NVP. The objective of this study was to determine whether decreasing iron exposure can mitigate NVP symptoms. Data were collected from a prospective cohort at the Motherisk Program in Toronto. Women (n = 97) seeking advice on managing severe NVP were advised to discontinue prenatal multivitamin administration and switch to folic acid, an adult multivitamin or a children's chewable multivitamin. Two-thirds (63 out of 97) (p < 0.001) of those women qualitatively reported an improvement in NVP symptoms after discontinuation of iron-containing prenatal multivitamins. These findings were verified quantitatively using both the pregnancy-unique quantification of emesis and nausea (PUQE) (p < 0.001) and well-being (p < 0.001) scoring systems. This is the first interventional study showing that discontinuation of iron results in improvement of NVP symptoms. Our data suggest that avoiding iron-containing prenatal multivitamins in the first trimester is effective in improving NVP symptoms in the majority of pregnant women suffering from morning sickness. PMID- 19280489 TI - Demographics and outcome of elective cerclage in a multi-ethnic London district general hospital. AB - Elective cerclage is a rare procedure, but is reported to be relatively more common in developing countries. This variation in rate may be reflected in a multi-ethnic population as seen in London. Our study analysed the epidemiology and rate of elective cerclage performed in a London district general hospital. Factors contributing to the outcome of the procedures were also reviewed. A total of 41 elective cerclages were performed in the hospital between 2000 and 2007. Data from these were collected retrospectively, including maternal history, operative details, and gestational age at delivery. Fisher's exact test was used for statistical analysis. Of the 41 cases, 19 pregnancies were carried to term (>or=37 weeks' gestation), nine were pre-term (24-36 weeks' gestation) and seven miscarried (<24 weeks' gestation); six cases had not yet delivered. All of the patients were immigrants from developing countries but ethnicity did not affect the operative outcome (p = 0.89, Fisher's exact). The other factors studied were also noted to have no significant impact on success. These included cervical length at insertion (p = 1.00, Fisher's exact), type of suture (p = 0.90, Fisher's exact) and average gestation at insertion (p = 0.20, Fisher's exact). In conclusion, all patients requiring intervention in this study originated from developing countries. This is a disproportionately high figure relative to the demographic breakdown of the study population. Such a finding may be due to geographical variation of risk factors for cervical incompetence but may also be influenced by observer bias. Additional studies are needed to further investigate the influence of ethnicity on the rate of elective cerclage. None of the variables analysed in this study significantly affected the outcome of the procedure. PMID- 19280490 TI - The role of recombinant activated Factor VII in major obstetric haemorrhage: the Farnborough experience. AB - Major obstetric haemorrhage is one of the commonest causes of maternal mortality and morbidity worldwide. It may result in coagulopathy and diffuse pelvic or vaginal bleeding. Correction of coagulopathy when administering Factor VII may be crucial to the management of selected cases. We report the use of recombinant activated factor in three cases of massive obstetric haemorrhage. Prolonged international normalised ratio, activated partial thromboplastin time, and reduced fibrinogen were the trigger to use rFVIIa. It was effective to halt the process of coagulopathy, secure haemostasis and improve laboratory parameters in all three patients. We review the relevant literature and discuss its indications, the potential benefits and possible complications. Recombinant activated Factor VII is a potential haemostatic agent in massive obstetric haemorrhage. Its successful use has been reported in post-surgical bleeding and consumptive coagulopathy. It may abolish the need for hysterectomy, which has a devastating effect on the patient future fertility and psychological well-being. PMID- 19280491 TI - General versus neuraxial anaesthesia for caesarean section: impact on the duration of hospital stay. AB - We investigated retrospectively the duration of hospital stay of 1,619 women who received general (GA) (n = 582) or neuraxial anaesthesia (combined spinal epidural [CSEA] (n = 614), epidural [EA] (n = 423)) for caesarean delivery over the years 2002-2005. Hospital stay was also analysed for the different obstetricians involved. Overall duration of hospital stay differed between 2002 and 2005 (p < 0.0001) but not between CSEA and EA (p = 0.460). Overall duration of hospital stay differed between neuraxial and GA group (p < 0.001). Duration of hospital stay of the GA group showed a progressive decrease between 2002 and 2005 (p = 0.002). Duration of hospital stay after neuraxial anaesthesia differed between 2002 and 2005 (p = 0.013) and among different surgeons (p < 0.001). Discharge rates from the hospital were shorter after neuraxial anaesthesia versus GA for the 3rd and 4th postoperative days (p < 0.001 and p < 0.001, respectively). Neuraxial anaesthesia for caesarean section seems to be associated with shorter duration of hospital stay than GA. PMID- 19280492 TI - Solifenacin for overactive bladder in women unsuccessfully treated with immediate release oxybutynin: a pilot study. AB - The aim of the study was to determine the efficacy, tolerability and cost comparison of solifenacin for women with an overactive bladder (OAB) who failed to respond to immediate release oxybutynin (IR). A standard 3-day bladder diary, cost of incontinence pads used over 4 weeks and a validated OAB quality-of-life questionnaire were collected at baseline, at 4 weeks and at 12 weeks of commencing solifenacin treatment. Nine women were enrolled into the study. Eight of the women completed the 12-week study and one woman withdrew. The mean number of day-time micturitions was 11.4 +/- 2.7 at baseline and 7.3 +/- 3.5 at 12 weeks of solifenacin treatment (p = 0.0002). The mean number of nocturia was 2.8 +/- 1.4 at baseline and 0.9 +/- 0.9 at 12 weeks of solifenacin treatment (p = 0.0004). The total number of incontinence episodes per day was 4.9 +/- 4.6 at baseline and 1.9 +/- 2.7 at 12 weeks of solifenacin treatment (p = 0.02). The mean micturition volumes were 160 +/- 50 ml at baseline and 280 +/- 50 ml at 12 weeks of solifenacin treatment (p = 0.002). The symptom severity domain of the OAB-questionnaire (OAB-q) showed a value of 60.8 +/- 23.0% at baseline and 32.0 +/- 25.9 % at 12 weeks of solifenacin treatment (p = 0.001). The health-related quality-of-life (HRQL) domain of the OAB-q showed a value of 45.5 +/- 28.0% at baseline and 73.3 +/- 24.8% at 12 weeks of solifenacin treatment (p = 0.0006). This study shows a significant improvement in bladder diary and validated quality of-life parameters with solifenacin in women with urge incontinence who have previously failed to respond to or have been intolerant of oxybutynin IR. PMID- 19280493 TI - Extracorporeal magnetic energy stimulation of pelvic floor muscles for urodynamic stress incontinence of urine in women. AB - The aim of this study was to assess the efficacy, side effects and drop out rate of extra-corporeal magnetic energy stimulation of pelvic floor muscles for urodynamic stress incontinence of urine in women. It was a prospective non controlled study at 2 district general hospitals in South Wales. It included 48 female patients with urodynamic stress incontinence of urine, who had 16, twice weekly treatment sessions. Pad test was the primary outcome measure and continence diary, King's Health and EuroQol quality of life questionnaires, side effects and drop out were the secondary outcome measures. Assessment was made on recruitment, at the end of treatment sessions and at 3 months follow up. Thirty one patients completed treatment sessions and 27 attended for follow up at 3 months. There was no significant change in outcome measures at the end of treatment or at 3 months follow up. Side effects were encountered by 52.1% of patients and the drop out rate was 35.4%. PMID- 19280494 TI - Trans-obturator tape: a single centre experience. AB - AIM: To assess the clinical outcome of TOT tape for stress and mixed urinary incontinence in a single centre. METHODS: From March 2002 to October 2006, 82 patients completed the study, all were evaluated at 3 and 12 months by physical examination and validated questionnaires. Seventy nine patients had the procedure under epidural anaesthesia and all women received antibiotics starting before surgery. RESULTS: TOT was mostly performed as a day case surgery with short operative time of 22 minute (range 15-38 minute). A total of 62 (70.4%) patients were discharged from the hospital within a few hours (4.3 +/- 1.7 hours). CONCLUSION: The TOT tape can safely be performed as a day-case procedure, which has a continence cure rate of approximately 80%. This figure is comparable with the more established TVT, however the TOT tape has a significantly lower morbidity in our experience. PMID- 19280495 TI - Patient perception of cervical screening among women living with human immuno deficiency virus infection attending an antiretroviral therapy clinic in urban South Africa. AB - This study aims to ascertain the perception of cervical screening practices among HIV-positive women attending an ART clinic in urban South Africa. It is a prospective cross-sectional study of 100 randomly selected patients using semi structured interviews. Answers to fixed-response questions were recorded for statistical analysis and themes were identified from responses to open-ended questions. The study found that 59% of women surveyed reported ever having had a Papanicolau (Pap) smear and that 41% of these women had never been notified of the result. Many women surveyed lacked understanding of cervical screening; 78% had never heard of cervical cancer and around 40% had no correct knowledge about Pap smears. The findings suggest that cervical screening practices among HIV positive women living in urban South Africa do not comply with the recommendations that are based on evidence of increased risk for this population. Systematic cervical screening programmes should be offered to HIV-positive women attending ART clinics in South Africa. PMID- 19280496 TI - An alternative to conventional sanitary protection: would women use a menstrual cup? PMID- 19280497 TI - Renal cell carcinoma in pregnancy. PMID- 19280498 TI - Elevated bile acids associated with acute hepatitis A infection in the third trimester of pregnancy. PMID- 19280499 TI - Prenatal diagnosis of massive fetal tongue teratoma followed by successful, ex utero intra-partum treatment. PMID- 19280500 TI - Prenatal cortical hyperostosis (Caffey disease) with Down syndrome. PMID- 19280501 TI - Unusual case of congenital chylothorax. PMID- 19280502 TI - Transvaginal ultrasonographic assessment of cervical length following mid trimester loss of one twin. PMID- 19280503 TI - To push? or not to push?: Second stage management in a patient with Stickler syndrome at risk of retinal detachment. PMID- 19280504 TI - Childbirth following previous repair of an imperforate anus. PMID- 19280505 TI - A novel case of amniotic fluid embolism in multiple pregnancy. PMID- 19280506 TI - Two cases of significant postoperative morbidity following peritoneal non-closure at caesarean section. PMID- 19280507 TI - The Shanghai ring: two case reports and review of literature. PMID- 19280508 TI - Pelvic haematoma and severe gluteal bruising associated with Ehler-Danlos syndrome following posterior colpoperineorrhaphy. PMID- 19280509 TI - Spontaneous proximal tubal stump pregnancy following partial salpingectomy. PMID- 19280510 TI - An unusual case of abdominal actinomycosis. PMID- 19280511 TI - Squamous cell carcinoma arising in ovarian mature cystic teratoma with isolated metastasis to the appendix. PMID- 19280512 TI - Primary Ewing's sarcoma--primitive neuroectodermal tumour of uterus. PMID- 19280513 TI - Leiomyosarcoma following uterine artery embolisation. PMID- 19280514 TI - Re: Mahran MA, Sayed AT, Imoh-Ita F. Avoiding over diagnosis of shoulder dystocia. Journal of Obstetrics and Gynaecology 2008;28(2):173-176. PMID- 19280517 TI - Drug-drug interaction studies in preclinical species: should metabolite(s) kinetics be studied? AB - Drug-drug interaction studies are important building blocks in drug development to understand the perceived risk of a purported interaction due to the differing clinical pharmacology attributes of the co-administered drugs. Two case studies are presented that justify the importance of evaluating the metabolite kinetics data along with the parent in a preclinical model. Atorvastatin and verapamil have interesting clinical pharmacology attributes in that both agents are substrates and/or inhibitors of the dual cytochrome P450 (CYP) 3A4 and P glycoprotein (Pgp) efflux transporter interplay. As articulated by the two case studies, the presence of metabolite kinetic data (i.e., norverapamil) provided unequivocal evidence in order to tease out the actual pathway responsible for the interaction between atorvastatin and verapamil. Therefore, consideration for metabolite kinetics, wherever feasible, appears to be prudent in defining the interaction liability between the two agents in a preclinical model. PMID- 19280518 TI - 2-ABT-S-oxide detoxification by glutathione S-transferases A1-1, M1-1 and P1-1: implications for toxicity associated with zileuton. AB - Zileuton, an agent which targets the leukotriene pathway through inhibition of 5 lipoxygenase (5-LO), was approved for the treatment of asthma in 1997. Shortly after its release, its use was restricted due to the observation of hepatotoxicity in patients. Previous research from the authors' laboratory demonstrated the formation of the reactive metabolite, 2-ABT-S-oxide (M1) from zileuton, and has identified a mercapturate of 2-ABT, C1, in the urine of rats dosed with zileuton. The reaction between M1 and glutathione (GSH) has been established in vitro; however, the potential for catalysis by glutathione transferases (GSTs) was not addressed. The work presented here outlines a role for GSTs in the detoxification of M1. Non-enzymatic conjugation studies with M1 and GSH in control experiments led to a t(1/2) of 6.4 +/- 0.4 h at pH 6.5. This rate was accelerated in the presence of GSTA1-1, GSTM1-1 and GSTP1-1 providing t(1/2) values of 2.6 +/- 0.1, 0.53 +/- 0.02, and 0.3 +/- 0.04 h, respectively, at pH 6.5. The inhibition of various GST enzymes was also studied. Results show that M1 inhibits GSTM1-1 and GSTP1-1 to a greater extent as compared with GSTA1-1. In the case of GSTA1-1, the inhibition was observed to be reversible, whereas M1 inhibition of GSTM1-1 and GSTP1-1 was found to be irreversible under identical conditions. GSTM1-1 is present in liver and thus the finding of the alkylation and potential irreversible inactivation of this isoform in vivo could contribute to an understanding of the hepatotoxicity associated with zileuton. PMID- 19280519 TI - Characterization of increased drug metabolism activity in dimethyl sulfoxide (DMSO)-treated Huh7 hepatoma cells. AB - The objective of this study was to characterize Huh7 cells' baseline capacity to metabolize drugs and to investigate whether the drug metabolism was enhanced upon treatment with dimethyl sulfoxide (DMSO). The messenger RNA (mRNA) levels of major Phase I and Phase II enzymes were determined by quantitative real-time polymerase chain reaction (RT-PCR), and activities of major drug-metabolizing enzymes were examined using probe drugs by analysing relevant metabolite production rates. The expression levels of drug-metabolizing enzymes in control Huh7 cells were generally very low, but DMSO treatment dramatically increased the mRNA levels of most drug-metabolizing enzymes as well as other liver-specific proteins. Importantly, functionality assays confirmed concomitant increases in drug-metabolizing enzyme activity. Additionally, treatment of the Huh7 cells with 3-methylcholanthrene induced cytochrome P450 (CYP) 1A1 expression. The results indicate that DMSO treatment of Huh7 cells profoundly enhances their differentiation state, thus improving the usefulness of this common cell line as an in vitro hepatocyte model. PMID- 19280520 TI - Comparison of formation of thiolactones and active metabolites of prasugrel and clopidogrel in rats and dogs. AB - Prasugrel and clopidogrel are antiplatelet prodrugs that are converted to their respective active metabolites through thiolactone intermediates. Prasugrel is rapidly hydrolysed by esterases to its thiolactone intermediate, while clopidogrel is oxidized by cytochrome P450 (CYP) isoforms to its thiolactone. The conversion of both thiolactones to the active metabolites is CYP mediated. This study compared the efficiency, in vivo, of the formation of prasugrel and clopidogrel thiolactones and their active metabolites. The areas under the plasma concentration versus time curve (AUC) of the thiolactone intermediates in the portal vein plasma after an oral dose of prasugrel (1 mg kg(-1)) and clopidogrel (0.77 mg kg(-1)) were 15.8 +/- 15.9 ng h ml(-1) and 0.113 +/- 0.226 ng h ml(-1), respectively, in rats, and 454 +/- 104 ng h ml(-1) and 23.3 +/- 4.3 ng h ml(-1), respectively, in dogs, indicating efficient hydrolysis of prasugrel and little metabolism of clopidogrel to their thiolactones in the intestine. The relative bioavailability of the active metabolites of prasugrel and clopidogrel calculated by the ratio of active metabolite AUC (prodrug oral administration/active metabolite intravenous administration) were 25% and 7%, respectively, in rats, and 25% and 10%, respectively, in dogs. Single intraduodenal administration of prasugrel showed complete conversion of prasugrel, resulting in high concentrations of the thiolactone and active metabolite of prasugrel in rat portal vein plasma, which demonstrates that these products are generated in the intestine during the absorption process. In conclusion, the extent of in vivo formation of the thiolactone and the active metabolite of prasugrel was greater than for clopidogrel's thiolactone and active metabolite. PMID- 19280521 TI - Utilization of estimated physicochemical properties as an integrated part of predicting hepatic clearance in the early drug-discovery stage: Impact of plasma and microsomal binding. AB - Rapid prediction of hepatic clearance for drug candidates plays an important role for decision-making in the early drug-discovery stage. Although knowledge of protein binding in both plasma and microsomal components is needed in the prediction of metabolic clearance from metabolic stability studies, the capacity of protein binding assays are generally lower than those of metabolic stability assays. However, many in silico prediction methods for protein binding are now available and software packages such as ACDLabs, ADMET Predictor and SimCYP incorporate various aspects of in silico predictions relevant to estimating binding and clearance. This has facilitated the use of various estimated or measured physicochemical parameters, relevant to binding, to predict clearance. In this study, prediction of protein binding for 33 drugs was evaluated using various combinations of estimated physicochemical properties. Subsequently, the most accurate estimated protein binding values were used to predict hepatic clearance using the SimCYP software. For the drugs used herein, SimCYP provided the most accurate prediction for protein binding in both plasma and microsomes using physiochemical properties estimated with the ACDLabs software. In conclusion, the use of in silico methods as an integrated part of predicting hepatic clearance in early drug-discovery stage is recommended. PMID- 19280522 TI - o-Nitroanilines as major metabolic products of anti-Trypanosoma cruzi 5 phenylethenylbenzofuroxans in microsomal and cytosolic fractions of rat hepatocytes and in whole parasitic cells. AB - The metabolism of six anti-Trypanosoma cruzi 5-phenylethenylbenzofuroxans (PhEBfx) was studied in vitro using rat hepatic microsomal and cytosolic fractions as a mammalian model and whole cells of T. cruzi as a parasitic model. Some of the expected metabolites were synthesized to provide authentic chromatographic standards. The metabolites were identified using high-performance liquid chromatography (HPLC) in comparison with the authentic standards and their proportions were determined. Their structures were confirmed using mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. The behaviour of the six PhEBfx in the three different systems was similar. The main metabolites, formed by reductive processes, were the corresponding o nitroanilines. Two of the test compounds were studied for extended time periods in the rat liver preparations and their terminal metabolites were identified as o phenylendiamine derivatives. PMID- 19280523 TI - Effect of Schisandra chinensis extract and Ginkgo biloba extract on the pharmacokinetics of talinolol in healthy volunteers. AB - The authors investigated the effect of herbal medicine Schisandra chinensis extract (SchE) and Ginkgo biloba extract (GBE) on the oral pharmacokinetics of P glycoprotein substrate talinolol in humans. Twelve healthy male volunteers took a single 100-mg oral dose of talinolol either alone or after pretreatment with 300 mg SchE twice daily or with 120 mg GBE three times daily for 14 days. On day 14, a single 100-mg oral dose of talinolol was administered. Plasma concentrations of talinolol from zero to 24 h were measured by high-performance liquid chromatography. SchE increased the area under the curve (AUC)(0-24) of talinolol by 47% (90% confidence interval (CI), 18-84%; p = 0.010), and GBE by 21% (90% CI = 11-32%; p = 0.002). The C(max) of talinolol increased by 51% (90% CI = 21-89%; p = 0.007) with SchE treatment and by 33% (90% CI = 18-51%; p = 0.002) with GBE treatment, respectively. The t(1/2) of talinolol increased by 7% (90% CI = -4% to 19%; p = 0.320) with SchE treatment and by 11% (90% CI = -12% to 38%; p = 0.436) with GBE treatment, respectively. The results suggest that both SchE and GBE significantly inhibited P-glycoprotein in humans. Patients receiving either SchE or GBE may require dose adjustments when treated with drugs primarily transported by P-glycoprotein. PMID- 19280524 TI - Plasma pharmacokinetics and metabolism of the antitumour drug candidate 2' benzoyloxycinnamaldehyde in rats. AB - The pharmacokinetics and metabolism of 2'-benzoyloxycinnamaldehyde (BCA) was characterized in male Sprague-Dawley rats as part of the preclinical evaluations for developing this compound as an antitumour agent. BCA was not detected in the plasma following either intravenous or oral dose, whereas its putative metabolites 2'-hydroxycinnamaldehyde (HCA) and o-coumaric acid were present at considerable levels. In separate pharmacokinetics studies, HCA exhibited a high systemic clearance and a large volume of distribution, whereas both pharmacokinetic parameters were much lower for o-coumaric acid. The terminal half life of both metabolites was approximately 2 h. BCA was converted rapidly to HCA in rat serum, liver microsomes and cytosol in vitro; HCA was subsequently converted to o-coumaric acid in a quantitative manner only in the liver cytosol. In addition, the formation of o-coumaric acid was inhibited significantly by menadione, a specific inhibitor for aldehyde oxidase. Taken collectively, the results suggest that the rapid systemic clearance of HCA is likely due mainly to hepatic clearance occurring from aldehyde oxidase-catalysed biotransformation to o- coumaric acid. In conclusion, the present work demonstrates that the anticancer drug candidate BCA is highly likely to work as its active metabolite HCA in the body. PMID- 19280525 TI - Exploration of the African green monkey as a preclinical pharmacokinetic model: oral pharmacokinetic parameters and drug-drug interactions. AB - African green monkeys (vervets) have been proposed as an alternate species that might allow improved access and provide high-quality pharmacokinetic results comparable with other primates. However, no oral data are available in vervets to evaluate cross-species predictive performance. Therefore, this study was conducted to evaluate the use of the vervet to predict human oral pharmacokinetics and drug interactions. Oral pharmacokinetic studies were conducted in the vervet for eight compounds: phenytoin, moxifloxacin, erythromycin, lidocaine, propranolol, ciprofloxacin, metroprolol, and prednisolone. To assess drug-drug interactions, co-administration experiments were conducted with ketoconazole and either propranolol or erythromycin. In general, the vervet provided similar predictivity for human oral exposure as cynomolgus or rhesus monkeys. In all non-human primates, human exposure to phenytoin would be over-predicted, and erythromycin, lidocaine, and propranolol under-predicted, with good predictivity for the other compounds studied. Furthermore, in the vervet, ketoconazole co-administration resulted in a six-fold increase in exposure to erythromycin, demonstrating proof of concept for drug drug interaction screening. These data support further exploration of the vervet as an alternate primate species for use in preclinical pharmacokinetic screening. PMID- 19280526 TI - Pharmacokinetics, tissue distribution, and metabolism of novel DNA topoisomerase I inhibitor yuanhuacine in rabbit. AB - A liquid chromatography/tandem mass spectrometry method was developed and validated for the quantitative determination of yuanhuacine (YHC), a daphne diterpene ortho-ester anticancer agent, and identification of its metabolites. Pharmacokinetic behaviour, tissue distribution, and metabolism were investigated in rabbit. YHC plasma data best fitted to a two-compartment model and were characterized by an elimination half-life t(1/2)(beta) of 11.1 h following intravenous administration. Tissue distribution studies did not identify any tissues having a high affinity for YHC. The main metabolites are proposed to be M392I, M392II, and M390, resulting from the ortho-ester group and aromatic ester bond being cleaved off simultaneously during Phase I metabolism. This investigation contributes to an understanding of the metabolism of daphne diterpene ortho-esters. PMID- 19280528 TI - Selected polysaccharides at comparison for their mucoadhesiveness and effect on precorneal residence of different drugs in the rabbit model. AB - Mucoadhesive polysaccharides may prolong the residence of ophthalmic drugs in precorneal area. In this article, the mucoadhesiveness of arabinogalactan, tamarind seed polysaccharide, hyaluronan, hydroxyethylcellulose is compared in vivo, by the polymer residence time in rabbit tear fluid, and in vitro, by the polymer-induced increase of viscosity of a mucin dispersion. Polymer residence is prolonged by increased viscosity but shortened by reflex tearing caused by excessive viscosity. Tamarind seed polysaccharide is the most effective in prolonging the residence of ketotifen and diclofenac in precorneal area; hence, it is the optimal eyedrop additive as it is mucoadhesive while not increasing viscosity excessively. PMID- 19280529 TI - High prevalence of esophageal involvement in lichen planus: a study using magnification chromoendoscopy. AB - BACKGROUND AND STUDY AIMS: The first cases of squamous cell carcinoma in esophageal lichen planus were recently described. We performed a study to establish the prevalence of endoscopic and histopathologic abnormalities consistent with lichen planus and (pre-) malignancy in a cohort of patients with lichen planus. PATIENTS AND METHODS: A total of 24 patients with lichen planus were prospectively studied using high-magnification chromoendoscopy. Focal esophageal abnormalities were mapped, classified, and biopsied. Biopsies were also taken from normal-appearing esophageal mucosa at three levels (proximal, middle, and distal). The presence of a lymphohistiocytic interface inflammatory infiltrate and Civatte bodies (i. e. apoptotic basal keratinocytes) at histopathologic examination was considered diagnostic for esophageal lichen planus. Symptoms were assessed using validated questionnaires. RESULTS: A total of 38 focal abnormalities were biopsied. These consisted of: layers of mucosa peeling off, hyperemic lesions, papular lesions, submucosal plaques/papules, a flat polypoid lesion, and segments of cylindrical epithelium. No endoscopic signs of dysplasia were present. Esophagitis consistent with gastroesophageal reflux disease was noted in 12 / 24 patients. Histopathology showed chronic inflammation of the esophageal mucosa in the majority (18 / 24) of patients. In 50 % (12 / 24), the diagnosis of esophageal lichen planus was made. Dysplasia was not present. There were no differences in symptoms between patients with and without esophageal lichen planus. CONCLUSIONS: At screening endoscopy a high prevalence (50 %) of esophageal lichen planus was found in patients with orocutaneous lichen planus. No dysplasia was found. PMID- 19280530 TI - The spiral enteroscopy training initiative: results of a prospective study evaluating the Discovery SB overtube device during small bowel enteroscopy (with video). AB - BACKGROUND AND AIM: Indications for small-bowel enteroscopy are increasing, but advancing the endoscope to the ileum remains challenging, especially for less experienced operators. The aim was to evaluate the ease of use, safety, and efficacy of the Discovery SB overtube (Spirus Medical, Stoughton, Massachusetts, USA) during SB enteroscopy by physicians with no experience of the device. PATIENTS AND METHODS: Thirty-three "untrained" endoscopists performed spiral enteroscopy during one of four 2-day training modules. Data were prospectively collected. Patient demographics, depth and time to maximal insertion, total procedure time, and findings were recorded. Trauma was documented during scope withdrawal. Day 1 and day 2 results were compared. RESULTS: Ninety procedures were successfully performed in 95 patients (72.6 % women, age = 48.8 +/- 14.2 years). Endoscopists each performed a mean of five procedures. Mean time to maximal insertion was 20.9 +/- 6.4 minutes. Mean depth achieved was 262.0 +/- 57.4 cm. Total procedure time was 33.6 +/- 8.0 minutes. In 90.3 %, 94.6 %, and 83.9 % of patients, respectively, a trauma score less than 3 was recorded in the esophagus, stomach, and intestine (scale = 0 - 5). There were no perforations, nor significant associations between trauma score and patient age, body mass index, depth of insertion, time to maximal insertion, total procedure time, or day 1 vs. day 2 procedures. Depth of insertion was greater on day 2 than on day 1 (276.9 +/- 53.7 cm vs. 252.0 +/- 58.0 cm, P = 0.043). CONCLUSIONS: Discovery SB provides safe advancement of the enteroscope into the distal small bowel. Maximum depth of insertion appears comparable to that of balloon enteroscopy while taking less time. The device is easy to use and may be effectively operated in as few as five training cases. PMID- 19280531 TI - Appropriateness of colonoscopy in Europe (EPAGE II). Screening for colorectal cancer. AB - BACKGROUND AND STUDY AIMS: To summarize the published literature on assessment of appropriateness of colonoscopy for screening for colorectal cancer (CRC) in asymptomatic individuals without personal history of CRC or polyps, and report appropriateness criteria developed by an expert panel, the 2008 European Panel on the Appropriateness of Gastrointestinal Endoscopy, EPAGE II. METHODS: A systematic search of guidelines, systematic reviews, and primary studies regarding colonoscopy for screening for colorectal cancer was performed. The RAND/UCLA Appropriateness Method was applied to develop appropriateness criteria for colonoscopy in these circumstances. RESULTS: Available evidence for CRC screening comes from small case-controlled studies, with heterogeneous results, and from indirect evidence from randomized controlled trials (RCTs) on fecal occult blood test (FOBT) screening and studies on flexible sigmoidoscopy screening. Most guidelines recommend screening colonoscopy every 10 years starting at age 50 in average-risk individuals. In individuals with a higher risk of CRC due to family history, there is a consensus that it is appropriate to offer screening colonoscopy at < 50 years. EPAGE II considered screening colonoscopy appropriate above 50 years in average-risk individuals. Panelists deemed screening colonoscopy appropriate for younger patients, with shorter surveillance intervals, where family or personal risk of colorectal cancer is higher. A positive FOBT or the discovery of adenomas at sigmoidoscopy are considered appropriate indications. CONCLUSIONS: Despite the lack of evidence based on randomized controlled trials (RCTs), colonoscopy is recommended by most published guidelines and EPAGE II criteria available online (http://www.epage.ch), as a screening option for CRC in individuals at average risk of CRC, and undisputedly as the main screening tool for CRC in individuals at moderate and high risk of CRC. PMID- 19280532 TI - Appropriateness of colonoscopy in Europe (EPAGE II). Surveillance after polypectomy and after resection of colorectal cancer. AB - BACKGROUND AND STUDY AIMS: To summarize the published literature on assessment of appropriateness of colonoscopy for surveillance after polypectomy and after curative-intent resection of colorectal cancer (CRC), and report appropriateness criteria developed by an expert panel, the 2008 European Panel on the Appropriateness of Gastrointestinal Endoscopy, EPAGE II. METHODS: A systematic search of guidelines, systematic reviews and primary studies regarding the evaluation and management of surveillance colonoscopy after polypectomy and after resection of CRC was performed. The RAND/UCLA Appropriateness Method was applied to develop appropriateness criteria for colonoscopy for these conditions. RESULTS: Most CRCs arise from adenomatous polyps. The characteristics of removed polyps, especially the distinction between low-risk adenomas (1 or 2, small [< 1 cm], tubular, no high-grade dysplasia) vs. high-risk adenomas (large [> or = 1 cm], multiple [> 3], high-grade dysplasia or villous features), have an impact on advanced adenoma recurrence. Most guidelines recommend a 3-year follow-up colonoscopy for high-risk adenomas and a 5-year colonoscopy for low-risk adenomas. Despite the lack of evidence to support or refute any survival benefit for follow-up colonoscopy after curative-intent CRC resection, surveillance colonoscopy is recommended by most guidelines. The timing of the first surveillance colonoscopy differs. The expert panel considered that 56 % of the clinical indications for colonoscopy for surveillance after polypectomy were appropriate. For surveillance after CRC resection, it considered colonoscopy appropriate 1 year after resection. CONCLUSIONS: Colonoscopy is recommended as a first-choice procedure for surveillance after polypectomy by all published guidelines and by the EPAGE II criteria. Despite the limitations of the published studies, colonoscopy is also recommended by most of the guidelines and by EPAGE II criteria for surveillance after curative-intent CRC resection. PMID- 19280533 TI - Appropriateness of colonoscopy in Europe (EPAGE II). Chronic diarrhea and known inflammatory bowel disease. AB - BACKGROUND AND STUDY AIMS: To summarize the published literature on assessment of appropriateness of colonoscopy for investigation of chronic diarrhea, management of patients with known inflammatory bowel disease (IBD), and for colorectal cancer (CRC) surveillance in such patients, and to report report appropriateness criteria developed by an expert panel, the 2008 European Panel on the Appropriateness of Gastrointestinal Endoscopy, EPAGE II. METHODS: A systematic search of guidelines, systematic reviews, and primary studies regarding the evaluation of chronic diarrhea, the management of IBD, and colorectal cancer surveillance in IBD was performed. The RAND/UCLA Appropriateness Method was applied to develop appropriateness criteria for colonoscopy for these conditions. RESULTS: According to the literature, colonoscopic evaluation may be justified for patients aged > 50 years with recent-onset chronic diarrhea or with alarm symptoms. Surveillance colonoscopy for CRC should be offered to all patients with extensive ulcerative colitis or colonic Crohn's disease of 8 years' duration, and to all patients with less extensive disease of 15 years' duration. Intervals for surveillance colonoscopy depend on duration of evolution, initial diagnosis, and histological findings. The EPAGE II criteria also confirmed the appropriateness of diagnostic colonoscopy for diarrhea of > 4 weeks' duration. They also suggest that, in addition to assessing extent of IBD by colonoscopy, further colonoscopic examination is appropriate in the face of persistent or worsening symptoms. Surveillance colonoscopy in IBD patients was generally appropriate after a lapse of 2 years. In the presence of dysplasia at previous colonoscopy, it was not only appropriate but necessary. CONCLUSIONS: Despite or perhaps because of the limitations of the available published studies, the panel-based EPAGE II (http://www.epage.ch) criteria can help guide appropriate colonoscopy use in the absence of strong evidence from the literature. PMID- 19280534 TI - Appropriateness of colonoscopy in Europe (EPAGE II). Iron-deficiency anemia and hematochezia. AB - BACKGROUND AND STUDY AIMS: To summarize the published literature on assessment of appropriateness of colonoscopy for the investigation of iron-deficiency anemia (IDA) and hematochezia, and report appropriateness criteria developed by an expert panel, the 2008 European Panel on the Appropriateness of Gastrointestinal Endoscopy, EPAGE II. METHODS: A systematic search of guidelines, systematic reviews and primary studies regarding the evaluation and management of IDA and hematochezia was performed. The RAND/UCLA Appropriateness Method was applied to develop appropriateness criteria for colonoscopy for these conditions. RESULTS: IDA occurs in 2 %-5 % of adult men and postmenopausal women. Examination of both the upper and lower gastrointestinal tract is recommended in patients with iron deficiency. Colonoscopy for IDA yields one colorectal cancer (CRC) in every 9-13 colonoscopies. Hematochezia is a well-recognized alarm symptom and such patients are likely to be referred for colonoscopy. Colonoscopy is unanimously recommended in patients aged > or = 50. Diverticulosis, vascular ectasias, and ischemic colitis are common causes of acute lower gastrointestinal bleeding (LGIB); CRC is found in 0.2 %-11 % of the colonoscopies performed for LGIB. Most patients with scant hematochezia have an anorectal or a distal source of bleeding. The expert panel considered most clinical indications for colonoscopy as appropriate in the presence of IDA (58 %) or hematochezia (83 %). CONCLUSION: Despite the limitations of the published studies, guidelines unanimously recommend colonoscopy for the investigation of IDA and hematochezia in patients aged > or = 50 years. These indications were also considered appropriate by EPAGE II, as were indications in patients at low risk for CRC with no obvious cause of bleeding found during adequate previous investigations. PMID- 19280535 TI - Appropriateness of colonoscopy in Europe (EPAGE II). Functional bowel disorders: pain, constipation and bloating. AB - BACKGROUND AND STUDY AIMS: To summarize the published literature on assessment of appropriateness of colonoscopy for the investigation of functional bowel symptoms, and report appropriateness criteria developed by an expert panel, the 2008 European Panel on the Appropriateness of Gastrointestinal Endoscopy, EPAGE II. METHODS: A systematic search of guidelines, systematic reviews and primary studies regarding the evaluation and management of functional bowel symptoms was performed. The RAND/UCLA Appropriateness Method was applied to develop appropriateness criteria for colonoscopy for these conditions. RESULTS: Much of the evidence for use of colonoscopy in evaluation of chronic abdominal pain, and/or constipation and/or abdominal bloating is modest. Major limitations include small numbers of patients and lack of adequate characterization of these patients. Large community-based follow-up studies are needed to enable better definition of the natural history of patients with functional bowel disorders. Guidelines stress that alarm features ("red flags"), such as rectal bleeding, anemia, weight loss, nocturnal symptoms, family history of colon cancer, age of onset > 50 years, and recent onset of symptoms should all lead to careful evaluation before a diagnosis of functional bowel disorder is made. EPAGE II assessed these symptoms by means of 12 clinical scenarios, rating colonoscopy as appropriate, uncertain and inappropriate in 42 % (5/12), 25 % (3/12), and 33 % (4/12) of these, respectively. CONCLUSIONS: Evidence to support the use of colonoscopy in the evaluation of patients with functional bowel disorders and no alarm features is lacking. These patients have no increased risk of colon cancer and thus advice on screening for this is not different from that for the general population. EPAGE II criteria, available online (http://www.epage.ch), consider colonoscopy appropriate in patients of > 50 years with chronic or new-onset bowel disturbances, but not in patients with isolated chronic abdominal pain. PMID- 19280536 TI - Appropriateness of colonoscopy in Europe (EPAGE II). Presentation of methodology, general results, and analysis of complications. AB - BACKGROUND AND STUDY AIMS: Appropriate use of colonoscopy is a key component of quality management in gastrointestinal endoscopy. In an update of a 1998 publication, the 2008 European Panel on the Appropriateness of Gastrointestinal Endoscopy (EPAGE II) defined appropriateness criteria for various colonoscopy indications. This introductory paper therefore deals with methodology, general appropriateness, and a review of colonoscopy complications. METHODS: The RAND/UCLA Appropriateness Method was used to evaluate the appropriateness of various diagnostic colonoscopy indications, with 14 multidisciplinary experts using a scale from 1 (extremely inappropriate) to 9 (extremely appropriate). Evidence reported in a comprehensive updated literature review was used for these decisions. Consolidation of the ratings into three appropriateness categories (appropriate, uncertain, inappropriate) was based on the median and the heterogeneity of the votes. The experts then met to discuss areas of disagreement in the light of existing evidence, followed by a second rating round, with a subsequent third voting round on necessity criteria, using much more stringent criteria (i. e. colonoscopy is deemed mandatory). RESULTS: Overall, 463 indications were rated, with 55 %, 16 % and 29 % of them being judged appropriate, uncertain and inappropriate, respectively. Perforation and hemorrhage rates, as reported in 39 studies, were in general < 0.1 % and < 0.3 %, respectively CONCLUSIONS: The updated EPAGE II criteria constitute an aid to clinical decision-making but should in no way replace individual judgment. Detailed panel results are freely available on the internet (www.epage.ch) and will thus constitute a reference source of information for clinicians. PMID- 19280538 TI - Use of a prokinetic agent after double-balloon endoscopy reduces abdominal distension and discomfort, and shortens recovery time: potential adjunct or alternative to CO2. PMID- 19280540 TI - Capsule endoscopy interpretation by an endoscopy nurse - a comparative trial. AB - BACKGROUND: Capsule endoscopy is a common, pain-free diagnostic procedure for the small bowel. However, interpretation of the whole video recording is a time consuming and costly procedure that can take up to 2 hours. The aim of the present study is two-fold: first to study the accuracy of capsule endoscopy analysis between a trained endoscopy nurse and a physician and secondly to determine if pre-evaluation by nursing staff might be time-effective for capsule reading. This study is especially important given the increasing financial pressure on current health-care systems. METHODS: A long-standing experienced endoscopy nurse, who was trained to read capsule endoscopy, and a physician, both blinded to the patient diagnosis and the other clinical findings reviewed 48 consecutive capsule endoscopy videos. The analyses of both the nurse and the physician were re-evaluated by an independent doctor regarding the agreement of the marked findings. RESULTS: Total time to read capsule endoscopy was significantly longer for the nurse's interpretation (63 +/- 26 min) as compared to the physician's interpretation (54 +/- 18 min, p < 0.01). The endoscopy nurse marked 236 thumbnails, whereas the doctor only marked 132 thumbnails. The nurse overlooked 4 of 64 relevant lesions (6 %), which had been detected by the physician. These overlooked lesions were not single important lesions, they were overlooked only in patients with multiple angiectasias of the small intestine, and thus the misdiagnosis was without clinical relevance. The physician overlooked 6 of 68 lesions detected by the nurse (9 %), also in patients with multiple angiodysplastic lesions and therefore without clinical relevance. On post-hoc analysis of the capsule video recordings the time needed by the physician to interpret the thumbnails marked by the nurse was 10 +/- 12 min. While there was no difference with respect to the estimated gastric emptying time (nurse 27 +/- 13 min vs. physician 28 +/- 14 min, n. s.), the estimated time of capsule passage through the ileocaecal valve was longer when interpreted by the endoscopy nurse (nurse 347 +/- 89 min vs. physician 326 +/- 74 min, n. s.). Nevertheless, the total cost for capsule pre-evaluation by the nurse was lower (13.23 euro vs. physician 17.82 euro). CONCLUSION: The endoscopy nurse detected 94 % of the significant lesions seen by the physician and no clinically relevant findings were overlooked. A pre-evaluation of the capsule video by trained staff is an accurate method and might be time effective. PMID- 19280541 TI - Impact of pantoprazole on duodeno-gastro-esophageal reflux (DGER). AB - BACKGROUND: Duodeno-gastro-esophageal reflux (DGER) is considered as an independent risk factor for complicated reflux disease (GERD). Patients with Barrett's esophagus have significantly higher levels of DGER than patients with uncomplicated GERD. However, the clinical response to conventional high-dose PPI therapy in patients with uncomplicated GERD and DGER is largely unknown. METHODS: 30 patients with uncomplicated GERD and combined pathological reflux (acid and bile) were enrolled in the study. Clinical work-up included evaluation of clinical symptoms, esophageal manometry and upper endoscopy. After 6 - 8 weeks of treatment with Pantoprazole 80 mg/d pH measurement and Bilitec 2000 were repeated, and the pattern of symptoms was re-evaluated. RESULTS: Under treatment with Pantoprazole 80 mg/d acid reflux was normalised in 28 patients (93 %). Similarly the mean percentage of DGER (time with an absorption greater than 0.14) was significantly reduced from 19.6 % (+/- 13.7) to 5.7 % (+/- 7.7, p < 0.05). In 15 patients (50 %) an elevated DGER persisted under treatment with Pantoprazole (DGER-NR group) whereas in 15 cases (50 %) a normalisation could be achieved (DGER-R group). The DGER-NR group had significantly higher levels of bile reflux before (and under) treatment compared to the DGER-R group: 22.9 % (9.98 %) vs. 15.6 % (0.72 %), respectively. Overall, the median quality of life index (QLI) improved from 4.78 (+/- 0.86) before to 8.04 +/- 1.84) under therapy. The clinical response under treatment was marikedly reduced in the DGER-NR group compared to the DGER-R group: QLI 7.3 vs. 8.9. Particularly heartburn and nocturnal coughing persisted. CONCLUSIONS: Our data confirm that high-dose pantoprazole therapy effectively exerts acid suppression in GERD patients with combined pathological reflux. However, DGER could only normalised in 50 % of patients. High levels of DGER at diagnosis enhance the risk of persistent DGER under PPI therapy and are associated with a reduced clinical outcome. PMID- 19280542 TI - A rare cause of ulcerative colitis - diarrhoea and perianal bleeding due to posttransplant lymphoproliferative disorder (PTLD). AB - Post-transplant lymphoproliferative disorder (PTLD) is characterised by frequent extranodal manifestation, in 20 - 25 % including the gastrointestinal tract. This entity, which is more frequent after solid organ transplantation, rarely occurs after bone marrow transplantation (BMT). We report the case of a 43-year-old male presenting with a short history of rectal bleeding, diarrhoea and weight loss. He had received a bone marrow transplant two years previously for an acute lymphocytic leukaemia of B-cell origin. On sigmoidoscopy, deep ulcerations of the rectal and sigmoideal mucosa were found. Further investigations revealed a diffuse infiltration of the liver, spleen, both kidneys and lungs. Histologically, a monomorphic post-transplant lymphoproliferative disorder was diagnosed, the subtype was a high grade diffuse-large cell Non-Hodgkin's lymphoma of B-cell origin. The biopsies showed that a major proportion of cells expressed Epstein-Barr virus encoded proteins typical for latent as well as lytic EBV infection. This is a common feature of PTLD and possibly plays a critical role in its pathogenesis. The current therapeutic approach to the subtype of PTLD we saw in this patient is CHOP chemotherapy, comprising the anti-CD 20 antibody rituximab if CD 20-positivity is present. This patient had a fatal course of the disease and died a few days after the first chemotherapy cycle due to severe multiple organ failure. PMID- 19280543 TI - [Thrombotic microangiopathy under an effective treatment with gemcitabine]. AB - A 67-year-old male patient with a pancreatic carcinoma and hepatic metastases was admitted with progressive dyspnea and anuria. Previously he had received five cycles of a palliative chemotherapy with gemcitabine and responded well with a reduction of the tumor mass. The laboratory results showed a distinct anemia, thrombocytopenia, an increase in creatinine and lactate dehydrogenase levels. With an additional finding of 21 per mille fragmentocytes in the periphery blood smear, we diagnosed a thrombotic microangiopathy (TMA). Until now the literature lacks well-defined therapy standards for this known but rare condition of gemcitabine. In a few case reports addressing the related microangiopathy of thrombotic thrombocytopenic purpura (TTP), which is characterised by a significant reduction of the von Willebrand factor (vWF) cleaving serum protease ADAMTS-13, encouraging results have been achieved by an immediate plasma exchange. Though ADAMTS-13 activity was not relevantly reduced in our patient, we still observed a rapid improvement of the clinical features as well as of LDH, thrombocytes and fragmentocytes under plasma exchange. The patient was discharged after one month in good clinical condition. Interestingly, during follow-up for further 21 months we found a continued stable status of the pancreatic carcinoma without any cytostatic therapy. In summary, this case provides evidence that the use of plasma exchange therapy in gemcitabine-associated TMA is justified. PMID- 19280544 TI - Splenic rupture in systemic amyloidosis - case presentation and review of the literature. AB - INTRODUCTION: Splenic rupture and hemorrhage into the abdominal cavity is an extremely unusual and rare complication of internal disease. Hemopathies, inflammatory or infiltrative diseases affecting the spleen are possible causes for such a complication. Splenomegaly is a factor, which may significantly increase the risk of rupture. CASE REPORT: The authors present the case of a 52 year-old patient with established systemic amyloidosis. In the past he had received treatment for undefined hepatic disease and anemia, which was established during the treatment of pathological vertebral fractures. Unfortunately, during the initial treatment he did not consent to a liver biopsy, which would have determined the etiology of the disease. Systemic infiltrative disease, affecting the spleen and other organs, was confirmed posthumously. Described are those forms of amyloidosis which usually represent an unfavorable course of the disease. CONCLUSION: Presented is a patient with a rare form of primary amyloidosis and fatal complication, spleenic rupture. PMID- 19280545 TI - [Terlipressin for hepatorenal syndrome - what is the role of plasma expansion and hemodynamic monitoring?]. PMID- 19280546 TI - [Indications for CT and MR arthrography--recommendations of the Musculoskeletal Workgroup of the DRG]. AB - The ongoing discussion about CT and MR arthrography is at least in part due to the lack of definite guidelines. The intention of the musculoskeletal workgroup of the DRG (Deutsche Rontgengesellschaft) was the establishment of recommendations for general guidance. After review of the recent literature, the indications for arthrographic examinations were discussed during a consensus meeting. Since the published data are insufficient and partially contradictory, no precise statements could be extracted from the literature. Therefore, the proposed recommendations are mainly based on expert opinions. In this review the main statements of the published literature are summarized and the recommendations of the musculoskeletal workgroup of the DRG are presented. PMID- 19280547 TI - [Reproducibility of readings of ISO C 3D and CT lumbar pedicle screw scans]. AB - PURPOSE: This study was conducted to evaluate the reproducibility of the reading of lumbar pedicle screw scans using a C-arm-based imaging system in comparison to computed tomography. The influence of the technique and the experience of the rater should be determined. MATERIALS AND METHODS: The lumbar spines of 23 patients were stabilized using 102 pedicle screws. The position of the screws was controlled intraoperatively using an Arcadis Orbic 3D scanner. All scans were evaluated independently by three raters. The position of the implants in reference to the pedicle walls was described. Additionally, another 100 lumbar pedicle screws in 16 patients were evaluated postoperatively with a multirow CT. Kappa according to Fleiss was calculated for the reproducibility of the rater statements. Each rater repeated the analysis of 24 screws to assess the intraobserver variance. RESULTS: The reports of the CT scans showed significantly less variation. The consent of all 3 raters was achieved in 79.4 vs. 65.1 % of cases. The Kappa values were 0.56 and 0.29, respectively. Poor results were obtained especially for the medial pedicle wall (consent 70.0 vs. 50.0 %). The influence of the experience of the rater was not able to be verified. CONCLUSION: The image quality of the ISO C 3D is worse than that of multirow CT scans for the evaluation of lumbar pedicle screws. This causes greater variance among the rater reports. We stopped using the ISO C 3D technique intraoperatively for the implantation of lumbar pedicle screws. PMID- 19280548 TI - Peripheral intravenous power injection of iodinated contrast media through 22G and 20G cannulas: can high flow rates be achieved safely? A clinical feasibility study. AB - PURPOSE: Modern examination protocols for computed tomography (CT) often require high injection rates of iodinated contrast media (CM). The purpose of this study was to evaluate the maximum achievable flow rates and stability of different peripheral intravenous catheters (IVC) in vitro and to assess the feasibility of higher injection rates through small IVC in vivo. MATERIALS AND METHODS: For in vitro experiments flow measurements followed by high pressure testing of different types of IVC (22, 20, and 18 gauge [G]) were performed. For the in vitro study 91 patients with already inserted 22 or 20G IVC who had been referred for CT received Iopamidol (300 mg iodine/ml) at flow rates between 2 and 5 ml/sec. Complications were documented. RESULTS: The maximal achievable flow rate of the tested IVC in vitro ranged from 5 to 8 ml/sec. No damage was observed during in vitro testing. The initially targeted in vivo flow rate was dropped in 33 of 91 (36 %) patients because the IVC could not be flushed adequately with saline before CM injection. Extravasation of CM occurred in 2 cases. In the remaining 58 patients the standard CT protocol was performed with flow rates of 3 ml/sec through 22G IVC and 5 ml/sec through 20G IVC, respectively. In this group, the extravasation of CM was observed twice (p > 0.05). CONCLUSION: Even with highly viscous CM, high flow rates can be applied in vitro in 22, 20, and 18G IVC without risking material damage. In vivo power injection of iodinated CM through 22G and 20G IVC seems to be safely achievable in the majority of patients with flow rates of up to 3 ml/sec and 5 ml/sec. Extravasation rates do not differ significantly between patients with high-flow or low-flow injections. PMID- 19280549 TI - [X-ray-induced DNA double-strand breaks after angiographic examinations of different anatomic regions]. AB - PURPOSE: The aim of this study was to investigate DNA double-strand breaks (DSBs) in blood lymphocytes as markers of the biological radiation effects in angiography patients. MATERIALS AND METHODS: The method is based on the phosphorylation of the histone variant H 2AX (gamma-H2AX) after formation of DSBs. Blood samples were collected before and up to 24 hours after exposure of 31 patients undergoing angiographies of different body regions. Blood lymphocytes were isolated, fixed, and stained with a specific gamma-H2AX antibody. Distinct foci representing DSBs were enumerated using fluorescence microscopy. Additional in-vitro experiments (10 - 100 mGy) were performed for evaluation of DBS repair. RESULTS: 15 minutes after the end of fluoroscopy values between 0.01 and 1.50 DSBs per cell were obtained. The DNA damage level normalized to the dose area product was 0.099 (cardiac angiographies), 0.053 (abdominal angiographies), 0.023 (pelvic/leg angiographies) and 0.004 excess foci/cell/mGym (2) (cerebrovascular angiographies). A linear correlation was found between gamma-H2AX foci levels and the dose area product (abdomen: R (2) = 0.96; pelvis/legs: R 2 = 0.71). In-vivo on average 46 % of DSBs disappeared within 1 hour and 70 % within 2.5 hours. CONCLUSION: gamma-H2AX immunofluorescence microscopy is a sensitive and reliable method for the determination of X-ray-induced DSBs during angiography. The DNA damage level depends on the dose, the exposed anatomic region, and the duration/fractionation of the X-ray exposure. PMID- 19280550 TI - [Ruptured giant myelolipoma of the adrenal gland with acute retroperitoneal hemorrhage]. PMID- 19280551 TI - Spiking expression of mu-crystallin mRNA during treatment with methimazole in patients with graves' hyperthyroidism. AB - mu-Crystallin is an NADPH-dependent cytosolic T3-binding protein. A knockout study in mice showed that mu-crystallin has a physiological function as a reservoir of T3 in the cytoplasm in vivo. Patients with nonsyndromic deafness were reported to have point mutations in the mu-crystallin gene. The expression of mu-crystallin is regulated by multiple factors. The present study was performed to determine whether thyroid function is related to the expression of mu-crystallin mRNA in peripheral mononuclear cells. We examined 23 normal healthy male and female subjects and 15 patients with Graves' disease. mu-Crystallin protein expression was determined immunohistochemically in peripheral mononuclear cells. The expression of mu-crystallin mRNA was assessed by reverse transcription of total RNA from peripheral mononuclear cells followed by quantitative PCR. mu Crystallin protein was detected in peripheral mononuclear cells. The mRNA expression was negatively correlated with age in normal female subjects. The values in female subjects were significantly higher than those in males. The values were positively correlated with serum TSH concentration. The values of the thyrotoxic patients with Graves' disease were lower than those in healthy subjects. A transient increase in mu-crystallin expression was observed within 14 42 days after the initial treatment with antithyroid medication. Thyroid hormone inversely relates to the expression of mu-crystallin mRNA in euthyroid mononuclear cells. Abrupt suppression of thyroid function leads to overexpression of mu-crystallin mRNA in thyrotoxic mononuclear cells. Thyroid hormone-regulated mu-crystallin expression may control thyroid hormone action via the intracytoplasmic T (3) capacity. PMID- 19280552 TI - Is contrast-enhanced US alternative to spiral CT in the assessment of treatment outcome of radiofrequency ablation in hepatocellular carcinoma? AB - PURPOSE: The present study was conducted to assess the efficacy of contrast enhanced ultrasound with low mechanical index in evaluating the response of percutaneous radiofrequency ablation treatment of hepatocellular carcinoma by comparing it with 4-row spiral computed tomography. MATERIALS AND METHODS: 100 consecutive patients (65 men and 35 women; age range: 62 - 76 years) with solitary hepatocellular carcinomas (mean lesion diameter: 3.7 cm +/- 1.1 cm SD) underwent internally cooled radiofrequency ablation. Therapeutic response was evaluated at one month after the treatment with triple-phasic contrast-enhanced spiral CT and low-mechanical index contrast-enhanced ultrasound following bolus injection of 2.4 ml of Sonovue (Bracco, Milan). 60 out of 100 patients were followed up for another 3 months. Contrast-enhanced sonographic studies were reviewed by two blinded radiologists in consensus. Sensitivity, specificity, NPV and PPV of contrast-enhanced ultrasound examination were determined. RESULTS: After treatment, contrast-enhanced ultrasound identified persistent signal enhancement in 24 patients (24 %), whereas no intratumoral enhancement was detected in the remaining 76 patients (76 %). Using CT imaging as gold standard, the sensitivity, specificity, NPV, and PPV of contrast enhanced ultrasound were 92.3 % (95 % CI = 75.9 - 97.9 %), 100 % (95 % CI = 95.2 - 100 %), 97.4 % (95 % CI = 91.1 - 99.3 %), and 100 % (95 % CI = 86.2 - 100 %). CONCLUSION: Contrast enhanced ultrasound with low mechanical index using Sonovue is a feasible tool in evaluating the response of hepatocellular carcinoma to radiofrequency ablation. Accuracy is comparable to 4-row spiral CT. PMID- 19280553 TI - Validity of the sonographic measurement of the diameters of the ascending aorta in rats. AB - PURPOSE: The objective of this investigation was to compare transthoracic ultrasound (US) determinations of ascending aortic diameters in rats with video microscopy (VM), the current standard for measuring aortic diameters in rats. MATERIALS AND METHODS: The diameter of the ascending aorta was measured in 111 adult Lewis male rats, by VM and US, with a 9 MHz probe, before and after intervention for induction of experimental aneurysm of the ascending aorta. RESULTS: The Bland-Altman test showed a high degree of agreement between the two methods, with a bias of only 0.23 mm (95 % confidence limits - 0.86 - 0.39 mm). Also, the measurements obtained by US correlated highly (r = 0.83, p < 0.0001) with those obtained by VM. Rat ascending aortic diameters obtained both by VM and US correlated significantly with the weight (r = 0.62 and r = 0.39, respectively), and with the age of the animals (r = 0.74 and r = 0.49, respectively). CONCLUSION: This study demonstrates that noninvasive US ascending aortic measurements are a reliable supplement to VM for the development of an ascending aortic aneurysm model, and for monitoring the efficiency of novel therapeutic agents. PMID- 19280562 TI - [Revista de Neurologia. Presentation]. PMID- 19280563 TI - [Advances in the diagnosis and subtyping of attention deficit hyperactivity disorder: what may lie ahead for DSM-V]. AB - INTRODUCTION AND AIMS: A number of problems have been identified through research and clinical practice with the current DSM-IV criteria for the diagnosis of attention deficit hyperactivity disorder (ADHD). This paper reviews some of these issues along with possible solutions for consideration in the construction of the criteria for DSM-V. DEVELOPMENT: Issues related to the length of symptom lists and how best to conceptualize the neuropsychological constructs they represent, differing developmental thresholds for diagnosis for adults vs. children and teens, the criterion for age of onset, problems related to the current approach to subtyping, and the development of new items for the adult stage of the disorder are discussed along with other issues pertinent to the continuing effort to test and revise the DSM criteria for ADHD as a function of ongoing empirical research. CONCLUSIONS: The present paper has briefly raised a number of issues that require some attention by the various workgroups charged with creating the DSM-V diagnostic criteria for ADHD. PMID- 19280564 TI - [Social skills in attention deficit hyperactivity disorder]. AB - INTRODUCTION AND DEVELOPMENT: Attention deficit hyperactivity disorder (ADHD) is a heterogeneous disorder that is often accompanied by poor social functioning and competence. Sixty to seventy per cent of children with ADHD are socially rejected and stigmatised because of their provocative, aggressive or disruptive behaviour. These patients have difficulty in monitoring and interpreting social cues, resolving interpersonal conflicts and in creating practical solutions. This has repercussions on their appreciation and relationships with their peers and educators, from the earliest pre-school ages even up to adulthood. Different hypotheses exist regarding its origins, ranging from the dysfunctional involvement of areas of the brain that play a part in executive and emotional functioning to perceptive defects in recognising emotions and failure in the socio-cognitive skills needed to perform competent social activity. CONCLUSIONS: Early diagnosis and pharmacological and psycho-educational intervention are essential for improving the cardinal symptoms of ADHD, as well as the social problems associated with or originated by it. PMID- 19280565 TI - [Prematurity: longitudinal analysis of executive functions]. AB - INTRODUCTION: Understanding cognitive development requires an interdisciplinary and neuropsychological approach. Executive functions facilitates cognitive activity and they are related to progressive cerebral configuration during pregnancy and infancy. AIM: One of the aims of the actual neuropsychology is the ontogeny of executive functions and their capacity to explain differential and normative developmental trends, specially because of its consequences on mental flexibility, monitoring, planning and cognitive control; they are also essential for good performance at school. The incidence of developmental risk factors as prematurity could affect long-term executive functioning expressed in learning difficulties or behavioral control. SUBJECTS AND METHODS: We studied, comparatively and longitudinally, the individual activity on objects displayed by typical babies (n = 25), and preterm babies (n = 10) from 1.5 to 2 years-old. RESULTS AND CONCLUSION: Applying systematic observational methodology, spontaneous babies' activity is registered. Double intra and inter-group analysis compare the data from the resolution of a non-verbal task through a multifaceted design. Results obtained show us differential pattern of early executive functioning among the groups studied. The growth of executive functioning is showed, too, through the ages studied for every group. PMID- 19280566 TI - [Training of executive function in preschool children with combined attention deficit hyperactivity disorder: a prospective, controlled and randomized trial]. AB - INTRODUCTION: School-age children with attention deficit hyperactivity disorder, combined type (ADHD-C) have executive function (EF) alterations. ADHD-C and EF alterations improve with methylphenidate or after training of working memory. AIM: To determine EF training effect on ADHD-C and EF alterations in preschoolers. SUBJECTS AND METHODS: INCLUSION CRITERIA: randomly assigned, 2-4 years old with ADHD-C, from 7-1-2004 to 6-30-2005, normal neurological examination, no evidences of underlying chronic disorders, no taking long term medications and parents agree to train or no EF daily. Parents filled out DSM-IV diagnostic criteria form for ADHD-C and children were tested with the Standard Dimensional Change Card Sort Task at the first visit and once a year from 1 to 3 years. Inattention, hyperactivity/impulsivity and EF average scores were statistically analyzed by the t Student for significance. RESULTS: Of 25, 13 were trained. ADHD-C incidence went down to 16 (64%), 6 (24%) and 10 (40%) trained and untrained. Incidence was 50, 40 and 25% 100, and 66.66 y 100 after 1, 2 and 3 years with and without training. Inattention average score was 8.25, 8.4 and 7 before and 6, 5.8, and 5.5 before and after 1, 2 and 3 years of training. Hyperactivity/impulsivity average score was 8.5, 8.4 and 7.75 before and 5.75, 5.6, and 5.25 after 1, 2 and 3 years of training. EF average score was 5.2, 4.8, and 5.5 before and 8.5, 10 and 9.5 after 1, 2 and 3 years of training. Degree of improving of inattention, hyperactivity/impulsivity and EF were statistically significant for training of EF for 1, 2 and 3 years (p = 0.013, 0.002, 0.0249). CONCLUSIONS: Children with preschool ADHD-C and EF disorders should receive training of EF for at least 3 years from age of diagnoses to improve their condition. PMID- 19280567 TI - [Sleep in neurodevelopmental disorders]. AB - INTRODUCTION: Sleep disorders can affect daytime functioning in a variety of neurodevelopmental disabilities as autism, attention deficit hyperactivity disorder and Asperger syndrome. DEVELOPMENT: Researchers using behavioural and/or electrophysiological measures have identified differences in sleep architecture among people with autism, attention deficit hyperactivity disorder and other developmental disorders. Children with neurodevelopmental disabilities have significant sleep problems, most commonly disorders of initiating and maintaining sleep. Studies of sleep disturbance in children with academic and behavioural problems have underscored the role that primary sleep disorders play in the clinical presentation of symptoms of inattention and behavioural dysregulation. CONCLUSIONS: Identification and treatment of sleep disorders in children with neurodevelopmental disabilities may improve daytime behaviour in this patients. When establishing a treatment plan, it is imperative to understand the underlying etiology of the sleep problem, which in many cases is multifactorial. PMID- 19280568 TI - [Oppositional defiant disorder: aspects related to sex differences and informant]. AB - INTRODUCTION: Oppositional defiant disorder (ODD) is one of the most common externalizing disorders in childhood. ODD prevalence global rates vary from 2% to 16%. Along with conduct disorder and attention deficit disorder, ODD is one of the leading reasons for referral to neuropediatric and psychiatric services. Even though ODD has recognized clinical importance, key aspects of its conceptualization and prevalence in childhood and adolescence remain uncertain. DEVELOPMENT: We examine previous research findings of ODD prevalence and analyze sex differences and differences according to informants. CONCLUSIONS: ODD prevalence rates present high variability. A number of studies suggest that ODD is more common in boys than in girls. Nevertheless, some authors point that this sex differences may be due to methodological bias. We recommend the validation of an ODD scale that has into consideration the following aspects: level of the subject's development (age), gender and environment. PMID- 19280569 TI - [Neuropsychological aspects of acquired diffuse brain injury in paediatric childhood]. AB - INTRODUCTION AND DEVELOPMENT: Acquired brain injury is one of the most common causes of long-term disability in childhood and traumatisms are the most prevalent cause. Diffuse axonal injury is associated with poor neuropsychological outcome. In contrast to early focal lesions, young age confers no advantage in the outcome of severe diffuse brain injury. Plasticity is incomplete for structural and functional deficits in children. The traditional view of enhanced reorganization of function after localized brain injury it can't be applied in early severe diffuse brain injury. In the paediatric population where the central nervous system is in a rapid state of growth, developmental factors may mask the difficulties and deficits in certain skills may not be evident until that skill is expected to emerge at a certain age. CONCLUSION: Like in adults the neuropsychological domains more affected in childhood are psychomotor speed, memory, attention and executive functioning. In childhood neuropsychological deficits will affect learning and adaptive behaviour. Sometimes deficits will be cumulative over time. PMID- 19280570 TI - [Infantile autism and mirror neurons]. AB - INTRODUCTION: Infantile autism is a disorder that is characterised by alterations affecting reciprocal social interactions, abnormal verbal and non-verbal communication, poor imaginative activity and a restricted repertoire of activities and interests. The causes of autism remain unknown, but there are a number of different approaches that attempt to explain the neurobiological causes of the syndrome. A recent theory that has been considered is that of a dysfunction in the mirror neuron system (MNS). DEVELOPMENT: The MNS is a neuronal complex, originally described in monkeys and also found in humans, that is related with our movements and which offers specific responses to the movements and intended movements of other subjects. This system is believed to underlie processes of imitation and our capacity to learn by imitation. It is also thought to play a role in language acquisition, in expressing the emotions, in understanding what is happening to others and in empathy. Because these functions are altered in children with autism, it has been suggested that there is some dysfunction present in the MNS of those with autism. CONCLUSIONS: Dysfunction of the MNS could account for the symptoms that are observed in children with autism. PMID- 19280571 TI - [A study of autism spectrum disorders and language disorders using magnetoencephalography. The scientific contribution of Dr. Munoz Yunta]. AB - INTRODUCTION AND DEVELOPMENT: Autism spectrum disorders (ASD) are characterised by the alteration of three basic areas of behaviour, qualitative alterations in reciprocal social interaction, qualitative alterations in communication and patterns of behaviour, and stereotyped, repetitive and restrictive activities and interests. Specific language impairment (SLI) is a disorder in which language is slow and retarded with respect to the patient's chronological age, and which is not related to a sensory-auditory or motor deficit or to a pervasive developmental disorder. In both disorders, epilepsy has an important role. Dr. Munoz Yunta's researches were focused in studying ASD and SLI using magnetoencephalography in order to look for epileptiform activity in precise cerebral areas. CONCLUSIONS: Patients with ASD showed specific epileptiform activity predominantly distributed in the perisylvian areas. In patients with Asperger's syndrome, epileptiform activity was mostly found in the right hemisphere. In patients with autism, no lateralized epileptiform activity was observed, although there was predominant activity in the left hemisphere. Subclinical epileptiform activity was found in the perisylvian areas in patients with ASD. Patients with SLI displayed epileptiform activity in the frontal and middle temporal regions of both hemispheres. Magnetoencephalography can be used to distinguish between patients with ASD and patients with SLI. PMID- 19280572 TI - [Asperger's syndrome and high-functioning autism: comorbidity with anxiety and mood disorders]. AB - INTRODUCTION: The comorbidity between Asperger's syndrome and affective and anxiety disorders seems to be meaningful. Therefore, it is presented a revision of the most recent scientific literature to provide empiric data with the aim to set out a research prospective. DEVELOPMENT: Assessment and diagnosis of the psychosocial performance confirm a meaningful proportion of cases with affective and anxious symptomatology, being converted into disorders some of them. CONCLUSION: It is necessary a better diagnosis because the affective and behavioural disturbances can be hidden as Asperger's syndrome and high functioning autism associated symptomatology. Identify and recognize this comorbidity will improve the psychosocial capacity of these persons. PMID- 19280573 TI - [Autism, epilepsy and temporal lobe pathology]. AB - INTRODUCTION: The social, language, and behavioral problems that occur with autism suggest that the syndrome affects a functionally diverse and widely distributed set of neural systems. The temporal lobe is an important part of the social brain, and manifests morphological and functional alterations in autism spectrum disorders (ASD). AIM. To describe the relationship between autism, epilepsy and temporal lobe dysfunction. DEVELOPMENT: Psychopathology is common in children with temporal lobe epilepsy, with over-representation of ASD and unusual disruptive behaviour disorders. Behaviors associated with damage to the amygdala and related temporal lobe structures in humans and nonhuman primates are strikingly similar to those seen in autism. The anatomic alterations observed in patients with temporal lobe epilepsy involve those structures responsible for social brain functioning, mainly amygdala, hippocampus and superior temporal sulcus. This is supported by studies demonstrating associations among temporal lobe epilepsy, ASD, and neuroimaging alterations in the hippocampus and amygdala. CONCLUSIONS: The complex relationship between autism and epilepsy, as reflected in the autism-temporal lobe epilepsy phenotype, provides a bridge to further knowledge of shared neuronal networks that can account for both the autisms and the epilepsies. There is a critical early stage of brain maturation during which temporal lobe epilepsy perturbs the development of brain systems that underpin social intelligence and possibly other cognitive skills, disrupting normal cortical organization and circuitry, thereby inducing an ASD. It's difficult to know if these patients became autistic because of repetitive epileptic seizures and/or persistent epileptiform activity on the EEG or because of the epileptogenic and psychopathological effects of temporal lobe lesions during early development (infancy and early childhood). PMID- 19280574 TI - [Early attention and specific intervention programs with autism spectrum disorders]. AB - INTRODUCTION: Autism spectrum disorder (ASD) is a developmental disorder that is characterised by qualitative deficiencies in social interaction and in communication, behaviour that is characterised by repetitive stereotyped patterns, and a restricted repertoire of interests and activities. DEVELOPMENT AND CONCLUSIONS: Early detection and diagnosis of this disorder by means of neuropaediatric techniques and procedures plays a decisive role in being able to set up an interdisciplinary therapeutic approach, in which specific early intervention programmes are noted for their effectiveness. The aim of these programmes is to attenuate or eliminate the alterations produced by ASD from the earliest stages of childhood development. But, in addition to attending to the child, interdisciplinary early intervention teams also work with the family and the environment the child lives in. This enables them to implement a joint response that allows for substantial improvements in the competencies that the child can acquire, as well as in the welfare and quality of life of the child and his or her family. PMID- 19280575 TI - [Communication and language: bidirectionality in interventions in children with autism spectrum disorder]. AB - INTRODUCTION: The relations between formal language skills and social skills are altered in a specific way in persons with autistic spectrum disorder (ASD) in a dimension that is very different to what occurs in other pathologies that affect the acquisition of such abilities. This should have implications in the models of intervention in the language of children with ASD. DEVELOPMENT: It is argued that with these children reference to 'normal' development is not sufficient: it is necessary to establish alternative models of language acquisition that take into account both the peculiarities of the cognitive, perceptive and social profile of children with ASD and the nature of the compensation mechanisms, whether they are spontaneous or induced. CONCLUSIONS: The development of language skills depends on the aptitudes for social communication, but it seems possible to take advantage of specifically linguistic skills (which are sometimes well preserved in children with ASD) to help improve social skills: this is what is meant by the bidirectionality that exists between language and communication. PMID- 19280576 TI - [Assessment of pragmatics from verbal spoken data]. AB - INTRODUCTION: Pragmatic assessment is usually complex, long and sophisticated, especially for professionals who lack specific linguistic education and interact with impaired speakers. AIM: To design a quick method of assessment that will provide a quick general evaluation of the pragmatic effectiveness of neurologically affected speakers. This first filter will allow us to decide whether a detailed analysis of the altered categories should follow. DEVELOPMENT AND CONCLUSIONS: Our starting point was the PerLA (perception, language and aphasia) profile of pragmatic assessment designed for the comprehensive analysis of conversational data in clinical linguistics; this was then converted into a quick questionnaire. A quick protocol of pragmatic assessment is proposed and the results found in a group of children with attention deficit hyperactivity disorder are discussed. PMID- 19280577 TI - [Dyslexia: a disease, a disorder or something else?]. AB - INTRODUCTION AND DEVELOPMENT: Although dyslexia is widely diagnosed and affects a very important portion of the childhood population, a conceptual definition of the condition is still lacking. This paper analyses different contributions from molecular genetics, neurocognitive research and evolutionary psychology so as to provide a series of reflections that may further our understanding of the nature of dyslexia and make it easier to categorise. Genetics has shown that dyslexia is linked to quantitative, pleiotropic genes. Moreover, dyslexia is not linked to a single gene, but is polygenic. Another genetic characteristic is its heterogeneity, that is to say, dyslexia may be due to a variety of different genetic combinations. This model, which has recently been put forward to make sense of and lend coherence to the data provided by genetics and neurocognitive research, has been called the multiple deficit model. Writing, on the other hand, is seen as a cultural technique that was invented only recently, if we compare it with whole process of human evolution. In order to learn to read and write successfully, the brain has had to adapt structures that were designed by the process of natural selection to perform functions that are typical of our species and have absolutely nothing to do with reading. CONCLUSIONS: Taking the multiple deficit model as its starting point, this article analyses the therapeutic response to stimulant medication as a way of improving nuclear aspects of dyslexia, when dyslexia and attention deficit hyperactivity disorder coincide. PMID- 19280578 TI - [Detection and specific studies in procedural learning difficulties]. AB - INTRODUCTION: The main disabilities in non-verbal learning disorder (NLD) are: the acquisition and automating of motor and cognitive processes, visual spatial integration, motor coordination, executive functions, difficulty in comprehension of the context, and social skills. AIMS. To review the research to date on NLD, and to discuss whether the term 'procedural learning disorder' (PLD) would be more suitable to refer to NLD. DEVELOPMENT: A considerable amount of research suggests a neurological correlate of PLD with dysfunctions in the 'posterior' attention system, or the right hemisphere, or the cerebellum. Even if it is said to be difficult the delimitation between NLD and other disorders or syndromes like Asperger syndrome, certain characteristics contribute to differential diagnosis. Intervention strategies for the PLD must lead to the development of motor automatisms and problem solving strategies, including social skills. CONCLUSIONS: The basic dysfunction in NLD affects to implicit learning of routines, automating of motor skills and cognitive strategies that spare conscious resources in daily behaviours. These limitations are partly due to a dysfunction in non-declarative procedural memory. Various dimensions of language are also involved: context comprehension, processing of the spatial and emotional indicators of verbal language, language inferences, prosody, organization of the inner speech, use of language and non-verbal communication; this is why the diagnostic label 'PLD' would be more appropriate, avoiding the euphemistic adjective 'non-verbal'. PMID- 19280579 TI - [Non-verbal learning disorder: cognitive-behavioural characteristics and neuropsychological aspects]. AB - INTRODUCTION AND DEVELOPMENT: Non verbal learning disorder is a neurological condition which is considered to be a developmental disability. It is characterised by a specific dysfunction in motor, visuospatial and social skills in patients with a normal intellect and development of language. Warning signs in school are poor psychomotor coordination, arithmetic skills and drawing activities. Social judgment and social problem solving are also typically impaired. Furthermore, these patients seem to have increasing risk of emotional disorders. CONCLUSIONS: Current evidence and theories suggest that dysfunction of white matter in the right hemisphere could be the cause. The clinical characteristics and the diagnostic criteria have not fully agreed on. A comprehensive neuropsychological evaluation reveals specific and heterogeneous patterns of difficulties for each child that would make possible to address and accommodate an educational program. PMID- 19280580 TI - [Non-verbal learning disorders: a clinical study and pharmacological treatment]. AB - INTRODUCTION: Non-verbal learning disorder (NVLD) is the name given to a complex group of difficulties with a well-defined biological substrate. Their clinical manifestations affect the motor area, visuospatial organisation and the social competencies. AIM: To analyse the semiology that differentially identifies NVLD by means of a model of interpretation based on the experience gained in a non verbal learning disorder service (NVLDS) and in the neuropaediatric unit of a hospital. PATIENTS AND METHODS: The study included 65 cases: 22 from a NVLDS (20 boys/2 girls), 17 between 6-11 years old and 5 between 12-16 years of age; and 43 cases from a hospital unit, with a mean age of 9.7 years (range: 8-16 years), of whom 59 were males (91%) and 6 were females (9%). The phenotype analysis was designed using a clinical inventory divided into four blocks (social, graphomotor, perceptive and attentional). Hospital sample: deficits in attention, motor control and perception (DAMP) (58%), graphomotor (18%), attentional (16%) and social (8%). NVLDS sample: graphomotor (35.8%), DAMP (29.8%), social (21%) and attentional (12%). RESULTS AND CONCLUSIONS: Clinical experience in NVLD is scarce. The early findings of this research encourage us to defend this 'NVLD continuum' model, in which the therapeutic response to psychopharmaceuticals is very similar to that of other processes, such as attention deficit hyperactivity disorder, although it does possess its own particularities especially with regard to distractibility and a misinterpreted paradoxical effect of methylphenidate. PMID- 19280581 TI - [The advantages of neuropsychological examination batteries in attention deficit hyperactivity disorder]. AB - INTRODUCTION: The neuropsychological assessment of the functions of attention and the executive functions in patients with clinical features compatible with a diagnosis of attention deficit hyperactivity disorder (ADHD) is considered a priority study to be able to offer a basic study. It is also necessary to be able to carry out a short- and long-term developmental follow-up of patients. DEVELOPMENT: An algorithm for diagnosing ADHD is proposed that includes both traditional and computerised tests for assessing the functions of attention and the executive functions. The development of new neuropsychological batteries for this purpose involves the use of several tests in computerised form that offer technical and methodological advantages as regards their use and the later treatment of the data obtained. CONCLUSIONS: The advantages of computerised assessment include management of answer times, minimum expression of the effect exerted by the researcher, savings in time, accurate and fast scoring, statistical management of new scores, and greater proximity to research programmes. PMID- 19280582 TI - [Attention deficit hyperactivity disorder in adults]. AB - INTRODUCTION AND DEVELOPMENT: Attention deficit hyperactivity disorder (ADHD) may exist in up to 60% of adults, whose first symptoms appeared before they were seven years old. Since it is a neurobiological condition, its basic symptoms lack of attention, hyperactivity and impulsiveness- are similar at all ages, but in adults the clinical manifestations are specific to the subgroup with more frequent comorbidities than in childhood. Manifestations that characteristically appear in adults are difficulty in concentrating, poorer memory and short-term memory, disorganisation, difficulties with self-discipline, impulsiveness, low self-esteem, mental restlessness, frustration and limited social skills. CONCLUSIONS: Today, adult cases are underdiagnosed because for a long time ADHD was thought to be a condition that only affected children and teenagers; however, although its clinical manifestations in adults are essentially similar to those seen in children, they are different and have different forms of presentation. There are no biomedical tests that allow an objective diagnosis to be established, and thus care must be taken to draw up and evaluate a patient record that includes possible symptoms since childhood, typical signs and symptoms, and any relevant comorbidities. Self-evaluation scales are necessary as the preferred instrument for use as an aid to the patient record. The main pharmacological treatment is based on psychopharmaceuticals such as immediate-, controlled- or extended-release methylphenidate and on atomoxetine; cognitive-behavioural and psychosocial treatment should also be associated. PMID- 19280584 TI - A novel computer-assisted drill guide template for lumbar pedicle screw placement: a cadaveric and clinical study. AB - BACKGROUND: The great accuracy of computer-assisted operative systems for pedicle screw insertion makes them highly desirable for spinal surgeries. However, computer-assisted pedicle screw placement is expensive, and the learning curve for these techniques is significant. We have developed a novel method of spinal pedicle stereotaxy by reverse engineering (RE) and rapid prototyping (RP) and have validated the method's accuracy by cadaveric and clinical study. METHODS: A volumetric CT scan was performed on each desired lumbar vertebra and a three dimensional (3D) reconstruction model was generated with MIMICS 10.1, while the optimal screw size and orientation were determined using UG Imageware 12.1. A drill template was created using UG Imageware 12.1, with a surface that is the inverse of the vertebral surface. The drill template and its corresponding vertebra were manufactured using RP. The method was tested on six cadavers without any fluoroscopic control at surgery. Eventually, the technology was applied in six clinical cases. RESULTS: The accuracy of the drill template was confirmed by preoperatively drilling the screw trajectory into the vertebra biomodel. In the cadaveric experiment, 36 pedicle screws were inserted and no pedicle perforation was observed by postoperative CT scan. In the six clinical patients, the best fit for positioning the template was easily found manually during the operation. The required time between fixation of the template to the lamina and insertion of the pedicle screw into each segment (one or two vertebrae) was 1-2 min. In total, 22 screws were inserted into T12-L5, with two to four screws/patient. No misplacement occurred using the individual templates. Fluoroscopy was used only once after all the pedicle screws had been inserted. The method significantly reduces operation time and radiation exposure for the members of the surgical team. CONCLUSIONS: The authors have developed a novel computer-assisted drill template for lumbar pedicle screw placement. This method has shown its ability to customize the placement and size of each screw, based on the unique morphology of the lumbar vertebra. The potential use of drill templates to place lumbar pedicle screws is promising. PMID- 19280585 TI - Application of computer-aided three-dimensional skull model with rapid prototyping technique in repair of zygomatico-orbito-maxillary complex fracture. AB - BACKGROUND: With the advent of CAD/CAM and rapid prototyping (RP), a technical revolution in oral and maxillofacial trauma was promoted to benefit treatment, repair of maxillofacial fractures and reconstruction of maxillofacial defects. METHODS: For a patient with zygomatico-facial collapse deformity resulting from a zygomatico-orbito-maxillary complex (ZOMC) fracture, CT scan data were processed by using Mimics 10.0 for three-dimensional (3D) reconstruction. The reduction design was aided by 3D virtual imaging and the 3D skull model was reproduced using the RP technique. In line with the design by Mimics, presurgery was performed on the 3D skull model and the semi-coronal incision was taken for reduction of ZOMC fracture, based on the outcome from the presurgery. RESULTS: Postoperative CT and images revealed significantly modified zygomatic collapse and zygomatic arch rise and well-modified facial symmetry. CONCLUSIONS: The CAD/CAM and RP technique is a relatively useful tool that can assist surgeons with reconstruction of the maxillofacial skeleton, especially in repairs of ZOMC fracture. PMID- 19280588 TI - Postoperative reduced dose of cisplatin concomitant with radiation therapy in high- risk head and neck squamous cell carcinoma. AB - BACKGROUND: The role of low doses of cisplatin and concomitant postoperative radiotherapy in high risk head and neck squamous cell carcinoma has not yet been defined. METHODS: Patients treated with definitive surgery, who had histological evidence of involvement of more than 2 lymph nodes, extracapsular extension of disease, perineural and/or intravascular invasion, involved or close surgical margins, received postoperative radiotherapy plus 75 mg/m(2) of cisplatin every 3 weeks during the radiotherapy cycle. The primary endpoints were to evaluate treatment compliance and overall, cause-specific, and disease-free survival. RESULTS: A total of 142 patients were enrolled. With a median follow-up of 40 months, 5-year overall survival was 68%, cause-specific survival 78% and disease free survival 82%. At multivariate analysis surgical margins status and extracapsular lymph node invasion were the only statistically significant prognostic factors. Fifty-three percent of the patients developed severe mucositis and 14% hematologic toxicity of grade 3. The 3 planned concomitant chemotherapy cycles were delivered to 48% of the patients. CONCLUSIONS: Postoperative radiotherapy and concomitant low-dose cisplatin was an effective treatment in high risk head and neck patients. The total toxicity observed was lower compared with that reported with higher doses of cisplatin, although the delivery of all the 3 planned chemotherapy cycles was challenging. The distant failure rate was high, which was an unsatisfactory result. PMID- 19280589 TI - Negative Helicobacter pylori status is associated with poor prognosis in patients with gastric cancer. AB - BACKGROUND: Recent studies have suggested that Helicobacter pylori (H. pylori) infection may be related to better prognosis in patients with gastric cancer, but to the authors' knowledge, this finding has not yet been validated. In the current study, the association between H. pylori status and clinical outcome was investigated in a large cohort of patients. METHODS: Frozen non-neoplastic gastric mucosa and serum samples obtained from 297 patients who underwent surgery for primary gastric cancer between 1988 and 2004 were retrieved from the serum and tissue bank of the study department. H. pylori status was defined by means of polymerase chain reaction (PCR) analysis for the vacA gene in gastric mucosa and by serologic assay of H. pylori and CagA antibodies. Univariate and multivariate analyses were used for the association between clinicopathologic variables and long-term outcome. RESULTS: Positivity for H. pylori infection was observed in 256 of 297 patients (86%), whereas in 41 patients (14%), PCR for vacA and both serologic tests were negative. Negative H. pylori status was found to be significantly associated with cardia location, advanced pT classification, noncurative surgery, and a lower 5-year survival rate after R0 resection (24% vs 57%; P < .001). Multivariate survival analysis confirmed H. pylori status as a significant prognostic factor (hazards ratio, 2.47; 95% confidence interval, 1.40 4.35 [P = .002]). The influence of H. pylori status on long-term survival was observed in patients with early as well as advanced pT classifications. CONCLUSIONS: Negative H. pylori status appears to be an indicator of poor prognosis in patients with gastric cancer, and is independent of other well-known clinical and pathologic prognostic variables. PMID- 19280590 TI - Involution of latent endometrial precancers by hormonal and nonhormonal mechanisms. AB - BACKGROUND: Inactivation of the PTEN suppressor gene has been shown to occur in the majority of endometrial cancer cases. Somatic PTEN inactivation by deletion and/or mutation, the first detectible change of endometrial carcinogenesis, has been reported to occur at a high frequency in the endometrium of normal premenopausal women, although few of these cases progress to cancer. It was hypothesized that the 50% to 60% reduced cancer risk attributed to oral contraceptives (OCPs) and intrauterine devices (IUDs) occurred in part through their activity as negative selection factors for these subclinical mutated glands. METHODS: A total of 71 women with a history of OCP use and 80 with a history of IUD use were age matched with 191 and 119 controls, respectively. Endometrial biopsy specimens were immunostained for PTEN, and each was scored for the presence or absence of PTEN-null glands (latent precancer). RESULTS: The frequency of latent precancers was found to be significantly reduced in OCP exposed (13%; odds ratio [OR], 0.19 [P < .001]) and IUD-exposed (18%; OR, 0.42 [P = .015]) women compared with respective matched controls (43% and 34%). The presence or absence of endometritis did not appear to be significantly correlated with PTEN status within the IUD-exposed group (P = .24). CONCLUSIONS: Normal appearing PTEN mutated endometrial glands, which are highly prevalent in the normal population, may be targets of endometrial cancer risk-modulating exposures. Some exposures reported to diminish the incidence of endometrial cancer in epidemiologic outcome studies, including OCP and IUD use, are associated with a proportionate decline in the frequency of latent precancers. Involution of pre-existing endometrial latent precancers, as evaluated by PTEN analysis, may provide an accessible surrogate marker for long-term endometrial cancer risk. PMID- 19280591 TI - Bleeding diathesis in patients with chronic myelogenous leukemia receiving dasatinib therapy. AB - BACKGROUND: The most frequent nonhematologic side effects associated with dasatinib therapy in patients with chronic myeloid leukemia (CML) are gastrointestinal, rash, and fluid retention syndromes. However, bleeding has been observed in some patients receiving dasatinib. In the current study, the authors investigated the risk factors and management of bleeding associated with dasatinib therapy for CML after imatinib failure. METHODS: The bleeding episodes associated with dasatinib therapy in 138 patients with CML who were consecutively treated at the study institution in clinical trials were evaluated. RESULTS: Bleeding occurred in 32 (23%) patients (grade >or=3 in 9 [7%] patients [according to National Cancer Institute Common Toxicity Criteria]), including in 12% of patients treated in chronic phase, 31% of patients treated in accelerated phase (AP), and 35% of patients treated in blast phase (BP) (P = .02). The majority of episodes (81%) affected the gastrointestinal tract. Basic coagulation studies were normal in 97% of patients who developed bleeding complications. Although 37% of episodes occurred with platelet counts >100 x 10(9)/L, multivariate analysis identified thrombocytopenia and advanced phase CML as risk factors for bleeding. A trend toward an increased risk with a twice-daily schedule was observed (P = .17). Management included dasatinib interruption for a median of 17 days (range, 3-51 days) in 47%, of patients and transfusions in 72% of patients. CONCLUSIONS: Bleeding occurs during dasatinib therapy, particularly in patients with AP or BP disease and low platelet counts. Appropriate clinical monitoring and the timely interruption of dasatinib therapy are warranted in this subset of patients. PMID- 19280592 TI - [(90)Yttrium-DOTA]-TOC response is associated with survival benefit in iodine refractory thyroid cancer: long-term results of a phase 2 clinical trial. AB - BACKGROUND: The authors aimed to explore the efficacy of (90)Yttrium-1,4,7,10 tetra-azacyclododecane N,N',N'',N'''-tetraacetic acid [(90)Y-DOTA]-Tyr(3) octreotide (TOC) in advanced iodine-refractory thyroid cancer. METHODS: In a phase 2 trial, the authors investigated biochemical response (assessed by serum thyroglobulin levels), survival, and the long-term safety profile of systemic [(90)Y-DOTA]-TOC treatment in metastasized iodine-refractory thyroid cancer. Adverse events were assessed according to the National Cancer Institute criteria. Survival analyses were performed by using multiple regression models. RESULTS: A total of 24 patients were enrolled. A median cumulative activity of 13.0 GBq (range, 1.7-30.3 GBq) was administered. Response was found in 7 (29.2%) patients. Eight (33.3%) patients developed hematologic toxicity grade 1-3, and 4 (16.7%) patients developed renal toxicity grade 1-4. The median survival was 33.4 months (range, 3.6-126.8 months) from time of diagnosis and 16.8 months (range, 1.8-99.1 months) from time of first [(90)Y-DOTA]-TOC treatment. Response to treatment was associated with longer survival from time of diagnosis (hazard ratio [HR], 0.17; 95% confidence interval [CI], 0.03-0.92; P = .04) and from time of first [(90)Y DOTA]-TOC therapy (HR, 0.20; 95% CI, 0.04-0.94; P = .04). The visual grade of scintigraphic tumor uptake was not associated with treatment response (odds ratio [OR], 0.98; 95% CI, 0.26-3.14; P = 1.00). CONCLUSIONS: Response to [(90)Y-DOTA] TOC in metastasized iodine-refractory thyroid cancer was associated with longer survival. Upcoming trials should aim to increase the number of treatment cycles. PMID- 19280593 TI - Crops with target-site herbicide resistance for Orobanche and Striga control. AB - It is necessary to control root parasitic weeds before or as they attach to the crop. This can only be easily achieved chemically with herbicides that are systemic, or with herbicides that are active in soil. Long-term control can only be attained if the crops do not metabolise the herbicide, i.e. have target-site resistance. Such target-site resistances have allowed foliar applications of herbicides inhibiting enol-pyruvylshikimate phosphate synthase (EPSPS) (glyphosate), acetolactate synthase (ALS) (e.g. chlorsulfuron, imazapyr) and dihydropteroate synthase (asulam) for Orobanche control in experimental conditions with various crops. Large-scale use of imazapyr as a seed dressing of imidazolinone-resistant maize has been commercialised for Striga control. Crops with two target-site resistances will be more resilient to the evolution of resistance in the parasite, if well managed. PMID- 19280598 TI - Crystal structure of human eIF5A1: insight into functional similarity of human eIF5A1 and eIF5A2. PMID- 19280599 TI - Shelling the Voronoi interface of protein-protein complexes reveals patterns of residue conservation, dynamics, and composition. AB - The accurate description and analysis of protein-protein interfaces remains a challenging task. Traditional definitions, based on atomic contacts or changes in solvent accessibility, tend to over- or underpredict the interface itself and cannot discriminate active from less relevant parts. We here extend a fast, parameter-free and purely geometric definition of protein interfaces and introduce the shelling order of Voronoi facets as a novel measure for an atom's depth inside the interface. Our analysis of 54 protein-protein complexes reveals a strong correlation between Voronoi Shelling Order (VSO) and water dynamics. High Voronoi Shelling Orders coincide with residues that were found shielded from bulk water fluctuations in a recent molecular dynamics study. Yet, VSO predicts such "dry" residues without consideration of forcefields or dynamics at a dramatically reduced cost. The interface center is enriched in hydrophobic residues. Yet, this hydrophobic centering is not universal and does not mirror the far stronger geometric bias of water fluxes. The seemingly complex water dynamics at protein interfaces appears thus largely controlled by geometry. Sequence analysis supports the functional relevance of dry residues and residues with high VSO, both of which tend to be more conserved. On closer inspection, the spatial distribution of conservation argues against the arbitrary dissection into core or rim and thus refines previous results. Voronoi Shelling Order reveals clear geometric patterns in protein interface composition, function and dynamics and facilitates the comparative analysis of protein-protein interactions. PMID- 19280600 TI - Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim. AB - Dihydrofolate reductase (DHFR) is the enzyme responsible for the NADPH-dependent reduction of 5,6-dihydrofolate to 5,6,7,8-tetrahydrofolate, an essential cofactor in the synthesis of purines, thymidylate, methionine, and other key metabolites. Because of its importance in multiple cellular functions, DHFR has been the subject of much research targeting the enzyme with anticancer, antibacterial, and antimicrobial agents. Clinically used compounds targeting DHFR include methotrexate for the treatment of cancer and diaminopyrimidines (DAPs) such as trimethoprim (TMP) for the treatment of bacterial infections. DAP inhibitors of DHFR have been used clinically for >30 years and resistance to these agents has become widespread. Methicillin-resistant Staphylococcus aureus (MRSA), the causative agent of many serious nosocomial and community acquired infections, and other gram-positive organisms can show resistance to DAPs through mutation of the chromosomal gene or acquisition of an alternative DHFR termed "S1 DHFR." To develop new therapies for health threats such as MRSA, it is important to understand the molecular basis of DAP resistance. Here, we report the crystal structure of the wild-type chromosomal DHFR from S. aureus in complex with NADPH and TMP. We have also solved the structure of the exogenous, TMP resistant S1 DHFR, apo and in complex with TMP. The structural and thermodynamic data point to important molecular differences between the two enzymes that lead to dramatically reduced affinity of DAPs to S1 DHFR. These differences in enzyme binding affinity translate into reduced antibacterial activity against strains of S. aureus that express S1 DHFR. PMID- 19280602 TI - Normal-mode-based modeling of allosteric couplings that underlie cyclic conformational transition in F(1) ATPase. AB - F(1) ATPase, a rotary motor comprised of a central stalk (gamma subunit) enclosed by three alpha and beta subunits alternately arranged in a hexamer, features highly cooperative binding and hydrolysis of ATP. Despite steady progress in biophysical, biochemical, and computational studies of this fascinating motor, the structural basis for cooperative ATPase involving its three catalytic sites remains not fully understood. To illuminate this key mechanistic puzzle, we have employed a coarse-grained elastic network model to probe the allosteric couplings underlying the cyclic conformational transition in F(1) ATPase at a residue level of detail. We will elucidate how ATP binding and product (ADP and phosphate) release at two catalytic sites are coupled with the rotation of gamma subunit via various domain motions in alpha(3)beta(3) hexamer (including intrasubunit hinge bending motions in beta subunits and intersubunit rigid-body rotations between adjacent alpha and beta subunits). To this end, we have used a normal-mode-based correlation analysis to quantify the allosteric couplings of these domain motions to local motions at catalytic sites and the rotation of gamma subunit. We have then identified key amino acid residues involved in the above couplings, some of which have been validated against past studies of mutated and gamma-truncated F(1) ATPase. Our finding strongly supports a binding change mechanism where ATP binding to the empty catalytic site triggers a series of intra- and intersubunit domain motions leading to ATP hydrolysis and product release at the other two closed catalytic sites. PMID- 19280603 TI - Effect of D to E mutation of the RGD motif in rhodostomin on its activity, structure, and dynamics: importance of the interactions between the D residue and integrin. AB - Rhodostomin (Rho) is a snake venom protein containing an RGD motif that specifically inhibits the integrin-binding function. Rho produced in Pichia pastoris inhibits platelet aggregation with a K(I) of 78 nM as potent as native Rho. In contrast, its D51E mutant inhibits platelet aggregation with a K(I) of 49 muM. Structural analysis of Rho and its D51E mutant showed that they have the same tertiary fold with three two-stranded antiparallel beta-sheets. There are no structural backbone differences between the RG[D/E] loop which extends outward from the protein core and the RG[D/E] sequence at its apex in a four-residue RG[D/E]M type I turn. Two minor differences between Rho and its D51E mutant were only found from their backbone dynamics and 3D structures. The R(2) value of E51 is 13% higher than that of the D51 residue. A difference in the charge separation of 1.76 A was found between the sidechains of positive (R49) and negative residues (D51 or E51).The docking of Rho into integrin alphavbeta3 showed that the backbone amide and carbonyl groups of the D51 residue of Rho were formed hydrogen bonds with the integrin residues R216 and R214, respectively. In contrast, these hydrogen bonds were absent in the D51E mutant-integrin complex. Our findings suggest that the interactions between both the sidechain and backbone of the D residue of RGD-containing ligands and integrin are important for their binding. PMID- 19280601 TI - Order propensity of an intrinsically disordered protein, the cyclin-dependent kinase inhibitor Sic1. AB - Intrinsically disordered proteins (IDPs) carry out important biological functions and offer an instructive model system for folding and binding studies. However, their structural characterization in the absence of interactors is hindered by their highly dynamic conformation. The cyclin-dependent-kinase inhibitor (Cki) Sic1 from Saccharomyces cerevisiae is a key regulator of the yeast cell cycle, which controls entrance into S phase and coordination between cell growth and proliferation. Its last 70 out of 284 residues display functional and structural homology to the inhibitory domain of mammalian p21 and p27. Sic1 has escaped systematic structural characterization until now. Here, complementary biophysical methods are applied to the study of conformational properties of pure Sic1 in solution. Based on sequence analysis, gel filtration, circular dichroism (CD), electrospray-ionization mass spectrometry (ESI-MS), and limited proteolysis, it can be concluded that the whole molecule exists in a highly disordered state and can, therefore, be classified as an IDP. However, the results of these experiments indicate, at the same time, that the protein displays some content in secondary and tertiary structure, having properties similar to those of molten globules or premolten globules. Proteolysis-hypersensitive sites cluster at the N terminus and in the middle of the molecule, whereas the most structured region resides at the C-terminus, including part of the inhibitory domain and the casein kinase-2 (CK2) phosphorylation target S201. The mutations S201A and S201E, which are known to affect Sic1 function, do not have significant effects on the conformational properties of the pure protein. PMID- 19280604 TI - Multiscale characterization of protein conformational ensembles. AB - We propose a multiscale exploration method to characterize the conformational space populated by a protein at equilibrium. The method efficiently obtains a large set of equilibrium conformations in two stages: first exploring the entire space at a coarse-grained level of detail, then narrowing a refined exploration to selected low-energy regions. The coarse-grained exploration periodically adds all-atom detail to selected conformations to ensure that the search leads to regions which maintain low energies in all-atom detail. The second stage reconstructs selected low-energy coarse-grained conformations in all-atom detail. A low-dimensional energy landscape associated with all-atom conformations allows focusing the exploration to energy minima and their conformational ensembles. The lowest energy ensembles are enriched with additional all-atom conformations through further multiscale exploration. The lowest energy ensembles obtained from the application of the method to three different proteins correctly capture the known functional states of the considered systems. PMID- 19280605 TI - Electrical signals polarize neuronal organelles, direct neuron migration, and orient cell division. AB - During early brain development, the axis of division of neuronal precursor cells is regulated tightly and can determine whether neurons remain in the germinal layers or migrate away. Directed neuronal migration depends on the establishment of cell polarity, and cells are polarized dynamically in response to extracellular signals. Endogenous electric fields (EFs) orient cell division and direct migration of a variety of cell types. Here, we show that cell division of cultured hippocampal cells (neuron-like cells and glial-like cells) is oriented strikingly by an applied EF, which also directs neuronal migration. Directed migration involves polarization of the leading neurite, of the microtubule associated protein MAP-2 and of the Golgi apparatus and the centrosome, all of which reposition asymmetrically to face the cathode. Pharmacological inhibition of Rho-associated coiled-coil forming protein kinases (ROCK) and phosphoinositide 3-kinase decreased, leading neurite orientation and Golgi polarization in the neurons in response to an EF and in parallel decreased the directedness of EF guided neuronal migration. This work demonstrates that the axis of hippocampal cell division, the establishment of neuronal polarity, the polarization of intracellular structures, and the direction of neuronal migration are all regulated by an extracellular electrical cue. PMID- 19280606 TI - Beyond early development: Xenopus as an emerging model for the study of regenerative mechanisms. AB - While Xenopus is a well-known model system for early vertebrate development, in recent years, it has also emerged as a leading model for regeneration research. As an anuran amphibian, Xenopus laevis can regenerate the larval tail and limb by means of the formation of a proliferating blastema, the lens of the eye by transdifferentiation of nearby tissues, and also exhibits a partial regeneration of the postmetamorphic froglet forelimb. With the availability of inducible transgenic techniques for Xenopus, recent experiments are beginning to address the functional role of genes in the process of regeneration. The use of soluble inhibitors has also been very successful in this model. Using the more traditional advantages of Xenopus, others are providing important lineage data on the origin of the cells that make up the tissues of the regenerate. Finally, transcriptome analyses of regenerating tissues seek to identify the genes and cellular processes that enable successful regeneration. Developmental Dynamics 238:1226-1248, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19280607 TI - Acetone, butanone, pentanone, hexanone and heptanone in the headspace of aqueous solution and urine studied by selected ion flow tube mass spectrometry. AB - Urine is commonly analysed in clinical practice by a variety of liquid-phase techniques to check for excessive ketone bodies, proteins and salts to name just a few compounds. However, little work has been carried out to measure the volatile compounds emitted by urine since these do not yet have an established role in clinical diagnosis. There is, however, a growing body of evidence that these volatile compounds can be indicators of adverse physiological conditions and disease and with the advent of sensitive gas-phase analytical methods they can be quickly quantified in urine headspace and potentially provide valuable support for clinical diagnosis. Thus, we are developing selected ion flow tube mass spectrometry, SIFT-MS, for the real-time analysis of urine headspace, ultimately to support rapid diagnosis in the clinical environment. In this paper we focus on volatile ketones in the headspace of aqueous solutions and urine donated by three healthy volunteers. Using SIFT-MS, we have unambiguously quantified in urine headspace acetone, by far the most abundant ketone, butanone, pentanone, hexanone and heptanone using NO(+) precursor ions. Further to this, we have determined the Henry's Law coefficients, HLC, for these ketones in aqueous solution to allow the liquid-phase concentrations in urine to be estimated from headspace levels of their vapours. In addition, the influence of the addition of physiological amounts of dissolved urea, sodium chloride and hydrochloric acid on the partitioning of these ketones between the aqueous phase and gas phase has been investigated and found to be small, which gives greater credence to the use of the HLC obtained using aqueous solutions for the estimation of ketone concentrations in urine. Finally, parallel measurements of the levels of acetone in exhaled breath and urine headspace have been obtained and shown to be very similar, which gives support to the previous deduction from breath analysis that acetone is a truly systemic compound. PMID- 19280608 TI - Matrix-assisted laser desorption/ionization by using femtosecond laser pulses in the near-infrared wavelength regime. AB - We observe a substantial matrix-assisted laser desorption/ionization (MALDI) signal when irradiating femtosecond laser pulses in the near-infrared spectral range centered around 800 nm and using standard MALDI matrices with absorption bands in the ultraviolet (UV) regime. The laser pulse energy dependence of this novel phenomenon is investigated in comparison with MALDI with near-UV laser pulses. Our observations show that multiphoton absorption/ionization could be a major issue among the MALDI processes when the sample is irradiated with ultra short laser pulses. PMID- 19280609 TI - Mass spectrometry of nanodiamonds. AB - Detonation nanodiamonds (NDs) were studied by time-of-flight mass spectrometry (TOF MS). The formation of singly charged carbon clusters, C(n) (+), with groups of clusters at n = 1-35, n approximately 160-400 and clusters with n approximately 8000 was observed. On applying either high laser energy or ultrasound, the position and intensity of the maxima change and a new group of clusters at n approximately 70-80 is formed. High carbon clusters consist of an even number of carbons while the percentage of odd-numbered clusters is quite low (< or =5-10%). On increasing the laser energy, the maximum of ionization (at n approximately 200 carbons) is shifted towards the lower m/z values. It is suggested that this is mainly due to the disaggregation of the original NDs. However, the partial destruction of NDs is also possible. The carbon clusters (n approximately 2-35) are partially hydrogenated and the average value of the hydrogenation was 10-30%. Trace impurities in NDs like Li, B, Fe, and others were detected at high laser energy. Several matrices for ionizing NDs were examined and NDs themselves can also be used as a matrix for the ionization of various organic compounds. When NDs were used as a matrix for gold nanoparticles, the formation of various gold carbides Au(m)C(n) was detected and their stoichiometry was determined. It was demonstrated that TOF MS can be used advantageously to analyze NDs, characterize their size distribution, aggregation, presence of trace impurities and surface chemistry. PMID- 19280610 TI - New aspects in fragmentation of peptide nucleic acids: comparison of positive and negative ions by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. AB - The use of peptide nucleic acids (PNAs) is steadily increasing in biochemistry and diagnostics. So far, PNAs have mostly been investigated using cationic conditions in mass spectrometry. Furthermore, the use of fragmentation techniques developed for peptides and proteins like infrared multiphoton dissociation (IRMPD) and electron capture dissociation (ECD) has barely been examined. However, especially the fragmentation behavior of PNA oligomers in negative ion mode is of high importance, due to the ability to interact with nucleic acids which are almost exclusively analyzed in the negatively charged state. In the current study PNA fragmentations under cationic and anionic conditions were investigated and different fragmentation techniques like collision-induced dissociation (CID), IRMPD and ECD were applied. Especially when using CID and IRMPD, amide bonds were broken, whereas ECD resulted in the elimination of nucleobases. Differences were also observed between positive and negative ionization, while the sequence coverage for the negative ions was superior to positive ions. The fragmentation behavior using IRMPD led to almost complete sequence coverage. Additionally, in anions the interesting effect of multiple eliminations of HNCO was found. PMID- 19280611 TI - Systematic characterization of the covalent interactions between (-) epigallocatechin gallate and peptides under physiological conditions by mass spectrometry. AB - (-)-Epigallocatechin gallate (EGCG) is a major bioactive component in leaves of green tea, and has been widely investigated for its anti-tumor activity. The interaction between EGCG and the key peptides or proteins (e.g. glutathione, enzymes) in vivo is thought to be involved in the toxicity and anti-cancer mechanism of EGCG. However, the true anti-tumor mechanism of EGCG is not clear, and few studies have focused on the reactivity of EGCG toward peptides or proteins under physiological conditions (pH 7.4, 37 degrees C). In this work, the covalent interactions between EGCG and model peptides containing one or more nucleophilic residues (i.e. Arg, Cys, Met, and alpha-NH(2) of the N-terminus of peptides) under physiological condition were fully characterized using mass spectrometry. It was found that EGCG can react with the thiol groups of peptides to form adducts under physiological conditions (pH 7.4, 37 degrees C), even in the absence of the peroxidase/hydrogen peroxide system. Besides the thiol groups of peptides, it is firstly reported that EGCG also reacts with alpha-NH(2) of the N-terminus or arginine residues of model peptides to form Schiff base adducts, and the methionine residues of model peptides can be easily oxidized by hydrogen peroxide (H(2)O(2)) generated during the process of EGCG auto-oxidation to form methionine sulfoxide products. The preference for the reaction of nucleophlic residues of peptides with EGCG was determined to have the following order: Cys > alpha-NH(2) of the N-terminus > Arg. The neutral loss ions of [M+H-170](+) and [M+H-138](+) were detected in all tandem mass spectra of the EGCG adducts of peptides, which indicates that these two neutral loss ions can be considered as the characteristic neutral loss ions of peptides modified by EGCG. Results of the present research provide insights into the toxicology and anti-tumor mechanism of EGCG in vivo. PMID- 19280612 TI - Doping control analysis of emerging drugs in human plasma - identification of GW501516, S-107, JTV-519, and S-40503. AB - An important aspect of preventive doping research is the rapid implementation of tests for emerging drugs with potential for misuse into routine doping control assays. New therapeutics of different classes such as PPARdelta-agonists (e.g. GW501516), ryanodine-calstabin-complex stabilizers (e.g. S-107 and JTV-519), and selective androgen receptor modulators (SARMs, e.g. S-40503) are currently used for the treatment of particular medical conditions such as metabolic syndrome, cardiac arrhythmia, debilitating diseases and osteoporosis, respectively. Due to their being at an early stage of clinical trials and the limited availability of data on the metabolism and possible renal elimination of the active drugs, the development of protocols for doping control analyses of plasma specimens could be an option for the detection of the circulating agents. The mass spectrometric fragmentation of four emerging drug candidates (GW501516, S-107, JTV-519, and S 40503) was elucidated by positive electrospray ionization and collision-induced dissociation using a high resolution/high accuracy mass spectrometer. A screening and confirmation procedure was established based on liquid chromatography/tandem mass spectrometry requiring a volume of 100 microL of plasma. Proteins were precipitated using acetonitrile, the specimens were centrifuged and the supernatant analyzed using a triple-quadrupole mass spectrometer employing multiple reaction monitoring of diagnostic ion transitions. The method was validated with regard to specificity, limits of detection (0.4-8.3 ng/mL), recoveries (72-98%), intraday and interday precisions (12-21%), and ion suppression/enhancement effects. PMID- 19280613 TI - From metacyclophanes to cyclacenes: synthesis and properties of [6.8]3cyclacene. AB - In this article we show synthetic pathways to [6.8](n)cyclacenes demonstrated by the de novo synthesis of [6.8](3)cyclacene as the first purely hydrocarbon cyclacene and of precursors for [6.8](4)cyclacene. The design of the de novo synthesis by exploring alternative pathways is discussed and various precursors are shown. Crucial to the synthesis of [6.8](3)cyclacene were two cyclization steps. The first is a Wittig trimerization reaction which yielded the hexamethyl substituted all-cis-[2(3)]metacyclophanetriene. For the second cyclization step the methyl groups were converted to aldehyde functionalities by two subsequent oxidation steps of N-bromosuccinimide (NBS) bromination and oxidation with 2 iodoxybenzoic acid (IBX). The final cyclization of the second set of double bonds was achieved by a McMurry-coupling reaction. Towards the synthesis of [6.8](4)cyclacene different synthetic pathways to methyl substituted all-cis [2(4)]metacyclophanetetraenes were explored. Insights into the structures of [2(3)]metacyclophanetri- and [2(4)]metacyclophanetetraenes were gained by X-ray crystallographic investigations on various intermediates. A crystallographic analysis of [6.8](3)cyclacene revealed a D(3h) symmetrical structure with planar benzene rings and a formation of tubular structures in the solid state. PMID- 19280614 TI - Synthesis, properties and formation of (RCp)Co- and (RCp)Rh stabilized[4.8]3cyclacene derivatives. AB - Metal-stabilized belts: A torus, 3, consisting of three four- and three eight membered conjugated rings and stabilized by (RCp)Co- and (RCp)Rh- units, was generated by irradiation of [(RCp)Co(CO)(2)] and [(RCp)Rh(C(2)H(4))(2)], respectively, and 1.Eleven metal stabilized [4.8](3)cyclacene derivatives were synthesized. The substances were prepared in one-pot reactions by irradiation of a solution of 5,6,11,12-tetradehydro-dibenzo[a,e]cyclooctatetraene and the corresponding cobalt reagents or rhodium compounds. The resulting cyclacene derivatives reveal D(3h) symmetry in solution. In the solid state the hoop shaped systems crystallize in layers, which are intercalated with solvent layers. To unravel the mechanism of the one-pot reaction we isolated an intermediate, which shows almost planar cyclooctatetraene rings. PMID- 19280615 TI - The intramolecular edge-to-face interactions of an aryl C-H bond and of a pyridine nitrogen lone-pair with aromatic and fluoroaromatic systems in some [3,3]metaparacyclophanes: a combined computational and NMR study. AB - Simple model systems based on the [3,3]metaparacyclophane skeleton have been designed to study the effect of fluorination of the "face" ring on the edge-to face (EtF) interactions with the C(Ar)-H bond of a phenyl ring or the nitrogen lone-pair of pyridine. Calculations established that in their more stable conformation the model systems adopt a tilted EtF disposition with the rim of the meta-substituted ring pointing towards the face of the para-substituted ring. Topomerization occurs by flipping of the meta-substituted ring, a process that involves the formation of an intermediate featuring an orthogonal EtF disposition of the arenes, which is less stable than the tilted arrangement. The energy barriers to the isomerization were determined by variable-temperature NMR spectroscopy and were well reproduced by DFT calculations. The variation in the energy barrier as a function of the substitution of the para-substituted ring could be rationalized well by a polar interpretation of the EtF interaction in the cases of model systems presenting the PyN...pi interaction but not in the cases of models featuring the C(Ar)-H...pi interaction. PMID- 19280616 TI - Allosteric switches: remote controls for proteins. AB - Long-distance service: Finding a selective small-molecule modulator for every domain of every human protein is a Herculean task. Wouldn't it be much easier to control the activity of any protein of choice by simply fusing it with a generally adaptable switch protein and controlling protein activity indirectly? PMID- 19280617 TI - Iron-mediated cleavage of C-C bonds in vicinal tricarbonyl compounds in water. AB - Three of a kind: Vicinal tricarbonyl compounds undergo C-C cleavage mediated by ferric ions (see scheme). The observed cleavage of ninhydrin and dehydroascorbic acid has relevance for amino acid detection and the metabolism of vitamin C. PMID- 19280618 TI - C(aryl)-O activation of aryl carboxylates in nickel-catalyzed biaryl syntheses. AB - Crucial breakthroughs in the activation of the C(aryl)-O bond of phenol derivatives were achieved almost simultaneously by two research groups (see scheme; Cy = cyclohexyl). Garg et al. coupled a range of aryl pivalates with arylboronic acids to give unsymmetrical biaryls. Shi et al. achieved this through C(aryl)-O activation of aryl carboxylates; the best results for the coupling of aryl boroxines were again obtained with aryl pivalates. PMID- 19280620 TI - Palladium-catalyzed three-component cascade cyclization reaction of bisallenes with propargylic carbonates and organoboronic acids: efficient construction of cis-fused bicyclo[4.3.0]nonenes. AB - A bicycle built for two: The title reaction affords cis-fused bicyclo[4.3.0]nonenes from readily available 1,5-bisallenes with structurally diverse propargylic carbonates and arylboronic acids (see scheme; X = NTs, C(E(1))(2) with E(1) = CO(2)Bn, SO(2)Ph, dba = trans,trans-dibenzylidenacetone). The reaction may involve a sequential oxidative addition, two different types of three carbopalladations, and a Suzuki-type coupling. PMID- 19280619 TI - Total synthesis of hopeahainol A and hopeanol. AB - Similar, but different: Possessing almost the same but not identical structures, the recently discovered natural products hopeahainol A (1) and hopeanol (2) exhibit important but differing biological properties (see structures). Their first total synthesis has now been achieved through a series of novel cascade reactions and skeletal rearrangements. PMID- 19280621 TI - Thoughts on the end of the article: the implications for nursing practice. PMID- 19280622 TI - Long-term monitoring shows hepatitis B virus resistance to entecavir in nucleoside-naive patients is rare through 5 years of therapy. AB - Patients with chronic hepatitis B virus (HBV) infection who develop antiviral resistance lose benefits of therapy and may be predisposed to further resistance. Entecavir (ETV) resistance (ETVr) results from HBV reverse transcriptase substitutions at positions T184, S202, or M250, which emerge in the presence of lamivudine (LVD) resistance substitutions M204I/V +/- L180M. Here, we summarize results from comprehensive resistance monitoring of patients with HBV who were continuously treated with ETV for up to 5 years. Monitoring included genotypic analysis of isolates from all patients at baseline and when HBV DNA was detectable by polymerase chain reaction (> or = 300 copies/mL) from Years 1 through 5. In addition, genotyping was performed on isolates from patients experiencing virologic breakthrough (> or = 1 log(10) rise in HBV DNA). In vitro phenotypic ETV susceptibility was determined for virologic breakthrough isolates, and for HBV containing novel substitutions emerging during treatment. The results over 5 years of therapy showed that in nucleoside-naive patients, the cumulative probability of genotypic ETVr and genotypic ETVr associated with virologic breakthrough was 1.2% and 0.8%, respectively. In contrast, a reduced barrier to resistance was observed in LVD-refractory patients, as the LVD resistance substitutions, a partial requirement for ETVr, preexist, resulting in a 5-year cumulative probability of genotypic ETVr and genotypic ETVr associated with breakthrough of 51% and 43%, respectively. Importantly, only four patients who achieved < 300 copies/mL HBV DNA subsequently developed ETVr. CONCLUSION: Long term monitoring showed low rates of resistance in nucleoside-naive patients during 5 years of ETV therapy, corresponding with potent viral suppression and a high genetic barrier to resistance. These findings support ETV as a primary therapy that enables prolonged treatment with potent viral suppression and minimal resistance. PMID- 19280623 TI - Dynamics and stability of amyloid-like steric zipper assemblies with hydrophobic dry interfaces. AB - Recent seminal investigations have suggested that the basic structural motif of amyloid fibers may be constituted by a tight association of two facing beta sheets (steric zipper). Although this model has been derived from crystal structures of small peptide models, several theoretical investigations, essentially focused on steric zipper interface containing large polar and/or aromatic side chains, have confirmed the stability of this motif in a crystal free context. To analyze the general validity of these findings, we carried out molecular dynamics (MD) simulations on aggregates stabilized by steric zipper interfaces made also of small or hydrophobic residues. In particular, we here characterized assemblies formed by the peptides SSTSAA and VQIVYK, whose structures have been recently solved at high resolution. In contrast to previous results obtained for polar/aromatic aggregates of the same size and with similar interface area, steric zipper assemblies composed of a pair of 10-stranded beta sheets show high fluctuations and significant distortions in the simulation timescales (40-60 ns). Taking into account the crystal packing, the effect of the addition of an extra sheet to the assemblies was also evaluated. The MD results indicate that this addition does not provide extra-stabilization to the pair of sheet models. Although present data do not preclude the possibility that the steric zipper association identified in the crystal structure is the basic motif of SSTSAA and VQIVYK fibers, our findings highlight the importance of the nature of residues directly involved in the motif. Indeed, polar and aromatic residues that may form intrasheet and intersheet interactions likely provide a strong contribution to the steric zipper motif stability. Along this line, assemblies endowed with hydrophobic residues presumably require larger interfaces. In line with this suggestion, MD analysis of the HET-s(218-289) prion models composed of a similar number of strands shows that the assembly is endowed with a remarkable stability. PMID- 19280624 TI - Effects of realism on extended violent and nonviolent video game play on aggressive thoughts, feelings, and physiological arousal. AB - Previous research has shown that playing violent video game exposure can increase aggressive thoughts, aggressive feelings, and physiological arousal. This study compared the effects that playing a realistic violent, unrealistic violent, or nonviolent video game for 45 min has on such variables. For the purpose of this study, realism was defined as the probability of seeing an event in real life. Participants (N=74; 39 male, 35 female) played either a realistic violent, unrealistic violent, or nonviolent video game for 45 min. Aggressive thoughts and aggressive feelings were measured four times (every 15 min), whereas arousal was measured continuously. The results showed that, though playing any violent game stimulated aggressive thoughts, playing a more realistic violent game stimulated significantly more aggressive feelings and arousal over the course of play. PMID- 19280625 TI - Perception of screening and risk reduction surgeries in patients tested for a BRCA deleterious mutation. AB - BACKGROUND: Women at a high risk for breast cancer are offered choices for screening or prophylactic surgeries. The aim of this study was to evaluate opinions regarding screening and surgical strategies in high-risk women. METHODS: Women at the authors' institution who received BRCA1 of 2 testing before July 2005 were sent a follow-up patient survey. The authors compared responses of women who tested positive for a deleterious mutation with those who tested negative. For those who expressed an opinion (agree vs disagree), a 2-sided Fisher exact test was used to compare responses. RESULTS: A total of 540 surveys were sent, and 312 were returned (58%). Of these, 217 had breast cancer, and 86 women tested positive for a mutation. No BRCA+ women felt mammograms were difficult to get because of discomfort, whereas 5.4% of the BRCA- group did (P = .039). Seventy percent of BRCA+ women agreed that prophylactic mastectomy (PM) is the most effective means for reducing risk, compared with 40% of BRCA- women (P < .001). PM was felt to be the only way to reduce worry in 64.7% of BRCA+ and in 34.4% of BRCA- women (P < .001). PM was felt to be too drastic for 36.1% of BRCA+ and 40.5% of BRCA- women (P = .562). Difficulty in deciding between screening and PM occurred in 23.9% of BRCA+ and 12.5% of BRCA- women (P = .046). After excluding women with bilateral breast cancers, 81.0% of women who agreed that PM was best to reduce risk underwent a PM versus 19.1% of those who disagreed (P < .001). Of women who felt PM was the only way to reduce worry, 84.2% underwent PM. Only 15.8% of women who did not believe that it was the only way to decrease worry underwent PM (P < .001). CONCLUSIONS: BRCA mutation carriers were more likely to believe PM to be the best way to reduce both risk and worry of breast cancer. High-risk women who agreed that PM was more likely to reduce risk and worry of breast cancer were more likely to proceed with this intervention. PMID- 19280626 TI - Triggered release from peptide-proteinoid microspheres. AB - Proteinoid microspheres (PM) are unusual polymers formed by the thermal condensation of amino acids. Although they have been studied for over 60 years, they are only now beginning to garner interest as controlled release agents. Although they are very biocompatible, it has been problematic to design useful triggers that release small molecules from PM interiors. This has severely limited their usefulness. In the present study, short peptides have been successfully incorporated into PMs during their formation. The resulting hybrid peptide-PMs can release their interior content when hydrolyzed by a proteinase. Specifically, if a matrix metalloproteinase (MMP) cleavage site peptide is incorporated into a PM, the peptide-PM will release interior contents only in the presence of the MMP recognizing the cleavage peptide. The release rate can be determined by the concentration of the peptide in the PM synthesis mixture. This potentially makes peptide-PMs useful for delivering inhibitors or drugs into acute and chronic wounds, periodontal disease sites, and other disease states involving the fine-tuned regulation of proteinases. PMID- 19280627 TI - Culture of human mesenchymal stem cells using electrosprayed porous chitosan microbeads. AB - The aim of this study was to fabricate porous chitosan microbeads using an electrospraying method into liquid nitrogen, then thawing and refreezing. The microbeads were then used to evaluate their potential for tissue engineering of human mesenchymal stem cells (hMSCs). Scanning electron microscopy (SEM) and a mercury porosimeter were used to show the morphology of the scaffolds formed and to determine their pore size and porosity. As the chitosan concentration increased (0.5, 1, 1.5, and 2 wt %), the diameter of the porous microbead increased from 350 to 890 microm, and the average pore size and the porosity decreased from 65 to 21 microm and 95 to 38%, respectively. The hMSCs were cultured onto the porous microbeads in a spinner flask. SEM images and methyl tetrazolium salt assays at 3, 7, 14, and 21 days of culture revealed that hMSCs had successfully attached and proliferated inside the porous microbeads. This study demonstrated that electrosprayed porous chitosan microbeads can be used as three-dimensional scaffolds for tissue engineering. PMID- 19280629 TI - Rheological blends for drug delivery. II. Prolongation of nerve blockade, biocompatibility, and in vitro-in vivo correlations. AB - Rheological polymer blends of hyaluronic acid (HA) and hydroxypropylmethyl cellulose (HPMC) were evaluated as prolonged duration delivery vehicles for local anesthetics using a rat sciatic nerve blockade model. HA-HPMC blends extended the duration of sensory block approximately threefold compared to that achieved using a bupivacaine solution. Blending HA and HPMC facilitated the injection of higher polymer concentration delivery vehicles and reduced the rate of polymer hydration compared to HA solutions, enabling prolonged drug release. The duration of effective nerve block was correlated with each of the zero shear viscosity, polymer concentration, yield stress, and gel point frequency of the blends, while a two-parameter model correlating duration of nerve block with zero shear viscosity and humectancy provided improved fits to the in vivo data compared to any single variable alone. The blends exhibited no cytotoxicity and induced only a mild short-term inflammatory reaction in vivo at the site of injection, with all blends largely resorbed 4 days postinjection. PMID- 19280630 TI - Recovery process of sciatic nerve defect with novel bioabsorbable collagen tubes packed with collagen filaments in dogs. AB - Autologous nerve graft is the most commonly applied treatment for the patients with peripheral nerve defect, while application is limited because of tissue availability and unfavorable donor site morbidity. To overcome this problem, peripheral nerve regeneration using a nerve conduit has been studied. Especially, nerve conduit using biodegradable materials has been considered promising. In this study, a potential of collagen nerve conduit has been studied with special reference to the regenerating process of a peripheral nerve. Twelve adult female Beagle dogs weighting 10-12 kg were used. The peroneal nerve was cut to make a 30 mm defect. The nerve defect was bridged by the collagen artificial nerve conduit. Comprehensive functional, electrophysiological, morphometrical, and histological analyses were performed until one year after operation. The wet weight of tibialis anterior muscles was only 32.4% of the healthy side at 24 weeks, which was recovered to 77.4% at 52 weeks after denervation. Electrophysiological evaluation of tibialis anterior muscle belly showed polyphasic wave at 52 weeks after implantation, which was almost half amplitude as compared with that of control. The diameters of myelinated nerve fibers thickened day by day, and the average diameter was 5.16 microm at PFN, 3.91 microm at CG, and 3.75 microm at DFN, and average thickness of myelin sheath was 0.94 microm at PFN, 0.46 microm at CG, and 0.55 microm at DFN after 52 weeks. The distribution of myelinated nerve fiber size in the 52 weeks group was distinctly bimodal with the major peak at approximately 2-4 microm and the minor peak at 10-12 microm. These findings were consistent with the distribution of the normal nerve fiber. This study proves the feasibility of the collagen artificial nerve conduit for promoting nerve regeneration, raises new possibilities of seeking alternatives to autograft for nerve repair. The results from this study showed detailed process of morphological, electrophysiological, and functional recovery of the regenerated nerve, which would provide scientific background for this novel therapy. PMID- 19280628 TI - Genetic polymorphisms in DNA repair genes as modulators of Hodgkin disease risk. AB - BACKGROUND: Although the pathogenesis of Hodgkin disease (HD) remains unknown, the results of epidemiologic studies suggest that heritable factors are important in terms of susceptibility. Polymorphisms in DNA repair genes may contribute to individual susceptibility for development of different cancers. However, to the authors' knowledge, few studies to date have investigated the role of such polymorphisms as risk factors for development of HD. METHODS: The authors evaluated the relation between polymorphisms in 3 nucleotide excision repair pathway genes (XPD [Lys751Gln], XPC [Lys939Gln], and XPG [Asp1104His]), the base excision repair XRCC1 (Arg399Gln), and double-strand break repair XRCC3 (Thr241Met) in a population of 200 HD cases and 220 matched controls. Variants were investigated independently and in combination; odd ratios (OR) were calculated. RESULTS: A positive association was found for XRCC1 gene polymorphism Arg399Gln (OR, 1.77; 95% confidence interval [95% CI], 1.16-2.71) and risk of HD. The combined analysis demonstrated that XRCC1/XRCC3 and XRCC1/XPC polymorphisms were associated with a significant increase in HD risk. XRCC1 Arg/Arg and XRCC3 Thr/Met genotypes combined were associated with an OR of 2.38 (95% CI, 1.24 4.55). The XRCC1 Arg/Gln and XRCC3 Thr/Thr, Thr/Met, and Met/Met genotypes had ORs of 1.88 (95% CI, 1.02-4.10), 1.97 (95% CI, 1.05-3.73), and 4.13 (95% CI, 1.50 11.33), respectively. XRCC1 Gln/Gln and XRCC3 Thr/Thr variant led to a significant increase in risk, with ORs of 3.00 (95% CI, 1.15-7.80). Similarly, XRCC1 Arg/Gln together with XPC Lys/Lys was found to significantly increase the risk of HD (OR, 2.14; 95% CI, 1.09-4.23). CONCLUSIONS: These data suggest that genetic polymorphisms in DNA repair genes may modify the risk of HD, especially when interactions between the pathways are considered. PMID- 19280631 TI - Long term in vivo degradation and tissue response to photo-cross-linked elastomers prepared from star-shaped prepolymers of poly(epsilon-caprolactone-co D,L-lactide). AB - Long term in vivo degradation, and tissue response to, cylindrical elastomers made of photo-cross-linked star-poly(epsilon-caprolactone-co-D,L-lactide) triacrylate were investigated through subcutaneous implantation in rats. The elastomers were prepared via UV initiated crosslinking of prepolymers of equimolar amounts of monomers; a high crosslink density elastomer (ELAST 1250) was prepared from a prepolymer of 1250 Da and a low crosslink density elastomer (ELAST 7800) was prepared from a prepolymer of 7800 Da. The elastomers were characterized using cross-polarization magic angle spinning solid state (13)C NMR and attenuated total reflectance fourier transform infrared spectroscopy. The progress of the in vivo degradation process was followed by employing SEM, uniaxial tensile, mass loss, water uptake, and sol content measurements. The rate of in vivo degradation was faster than the rate of in vitro degradation for both ELAST 1250 and ELAST 7800. Long term in vivo degradation studies indicated that both elastomers undergo bulk hydrolysis along with surface erosion occurring due to the physiological environment. In the case of low cross-link density elastomers, the onset of mass loss was accompanied with an increase in both water uptake and sol content, whereas, in the case of high crosslink density elastomers, only the water uptake increased. This degradation pattern was due to crazing of the high crosslink density elastomers. ELAST 7800 cylinders were totally degraded, and ELAST 1250 cylinders had lost 80% of their mass, within 30 weeks. Aminor host reaction with minimal vascularity and inflammation was invoked, with a milder tissue response observed with more highly crosslinked cylinders. PMID- 19280632 TI - Formulation and immunological evaluation of novel vaccine delivery systems based on bioresorbable poly(ethylene glycol)-block-poly(lactide-co-epsilon caprolactone). AB - Novel emulsion-type vaccine delivery systems based on the amphiphilic bioresorbable polymer poly(ethylene glycol)-block-poly(lactide-co-epsilon caprolactone) (PEG-b-PLACL) and selected oils were developed here. Physicochemical characterizations such as stability, a droplet test, microscopic aspects, and in vitro release showed that PEG-b-PLACL-emulsified formulations have several advantages over traditional vaccine adjuvants in that they are stable, reproducible, and homogeneous fine particles with an appropriate size to facilitate the induction of potent immune responses. Different dispersion-type emulsions have provided different release profiles using ovalbumin in model studies. Immunogenicity studies in mice have shown that antigen-specific antibody titers and T-cell proliferative responses, as well as the secretion of IFN-gamma, were significantly enhanced for ovalbumin after formulation with PEG-b-PLACL based emulsions. These features are of great interest for applications in delivery systems of prophylactic and therapeutic vaccine candidates. PMID- 19280633 TI - Bone responses to rough titanium implants coated with biomimetic Ca-P in rabbit tibia. AB - A new technique to biomimetic deposition calcium phosphate (BDCaP) coating onto rough titanium substrates has been developed recently. This biomimetic deposition technique seems to be promising. It appears to have some advantages such as an inexpensive and simple set-up, and the possibility to synthesize layers with a defined surface morphology. The aim of this study was to examine the bone responses to BDCaP-coated implants in a rabbit model. Thirty one implants (16 BDCaP and 15 rough) were inserted into both tibia of 15 rabbits. After 2, 4, and 8 weeks following the implantation, the tibias were retrieved and prepared for histological evaluation. After 2 weeks, BDCaP-coated implant showed more bone ingrowth inside threads than the rough implants in medullary region (31.43% vs. 24.38%). Histological and quantitative histomorphometrical measurements demonstrated no more bone ingrowth and bone-implant contact for coated implant as compared with uncoated implant in cortical region at all experiment periods. From the histological viewpoint, the BDCaP coating did not have any positive effect on new bone formation. PMID- 19280634 TI - Effects of mechanical stimuli on the synthesis of superficial zone protein in chondrocytes. AB - Superficial zone protein (SZP) has been demonstrated to contribute to the boundary lubrication in synovial joints. This study was designed to clarify the modulation of SZP expression by mechanical stress in articular chondrocytes. Cyclic tensile strains of 7 and 21% cell elongation were applied to cultured chondrocytes obtained from porcine mandibular condylar cartilage. The mRNA levels of SZP, IL-1 beta, and TGF-beta1 were examined by a quantitative real-time PCR analysis. Protein level of SZP was examined by Western blotting. The SZP mRNA level was significantly upregulated after 12, 24, and 48 h by 7% elongation. Although SZP mRNA level was upregulated by 21% elongation after 12 h, it decreased to a lower level than the control after 48 h. The TGF-beta1 mRNA level exhibited an almost similar change to SZP. The IL-1 beta mRNA level was not changed markedly by 7% elongation. However, the IL-1 beta mRNA level was significantly increased by a 12-h application of 21% elongation. Western blot analysis revealed that the SZP expression was increased by 7% elongation, but decreased remarkably by 21% elongation. It is suggested from these findings that the SZP expression level in the chondrocytes is enhanced by optimal mechanical stimuli, but inhibited by excessive loading partly affected by TGF-beta1 and IL-1 beta, leading to the deterioration of joint lubrication. PMID- 19280635 TI - Proliferation and osteogenesis of immortalized bone marrow-derived mesenchymal stem cells in porous polylactic glycolic acid scaffolds under perfusion culture. AB - Human bone marrow mesenchymal stem cells (hMSCs) are promising candidates for cell therapy and tissue engineering. However, the life span of hMSCs during in vitro culture is limited. Human telomerase catalytic subunit (hTERT) gene transduction could prolong the life span of hMSCs and maintain their potential of osteogenic differentiation. Therefore, hMSCs transduced with hTERT (hTERT-hMSCs) could be used as a cell model for in vitro tissue engineering experiment because of its prolonged life span and normal cellular properties. A perfusion culture system for proliferation and osteogenesis of hTERT-hMSCs or primary hMSCs in porous polylactic glycolic acid (PLGA) scaffolds is described here. A cell suspension of hTERT-hMSCs or primary hMSCs (5 x 10(5) cells/250 microL) was seeded and then cultured for 12 days in porous PLGA scaffolds (10 mm in diameter, 3 mm in height) under both static and perfusion culture systems. The seeding efficiency, proliferation, distribution and viability, and osteogenesis of cells in scaffolds were evaluated. The perfusion method generated higher scaffold cellularity and proliferation of cells in scaffolds, and hTERT-hMSCs showed the higher proliferation potential than primary hMSCs. Results from fluorescein diacetate (FDA) staining and scanning electron microscopy (SEM) demonstrated homogeneous seeding, proliferation, and viability of hTERT-hMSCs throughout the scaffolds in the perfusion culture system. On the contrary, the static culture yielded polarized proliferation favoring the outer and upper scaffold surfaces, and resulted in decreasing of cells in the central section of the scaffolds. A flow rate of 0.5 mL/min had an effect on osteogenic differentiation of cells in scaffolds. However, the osteogenic medium promoted the osteogenic efficiency of cells. Scaffolds with hTERT-hMSCs had the higher osteogenesis than scaffolds with primary hMSCs. Thus, these results suggest that the flow condition not only allow a better seeding efficiency and homogeneity but also facilitate uniform proliferation and osteogenic differentiation of hTERT-hMSCs in scaffolds. hTERT hMSCs could be used as stem cell candidates for bone tissue engineering experiments. PMID- 19280636 TI - Chondrogenic differentiation of mesenchymal stem cells embedded in a scaffold by long-term release of TGF-beta 3 complexed with chondroitin sulfate. AB - In this study, mesenchymal stem cells (MSCs) embedded in biodegradable and water swollen, elastic block copolymer scaffolds were assessed for MSC chondrogenesis. To determine the optimal conditions for chondrogenesis of the embedded rMSCs, transforming growth factor-beta 3 (TGF-beta 3) was physically conjugated with chondroitin sulfate (CS) and mixed into scaffolds, which were subsequently evaluated for the differentiation of transplanted rMSCs. In determination of CS bound growth factors for chondrogenesis, scaffold mixed with rMSCs and TGF-beta 3 was then tested by growth factor release profiles, confocal laser microscopy, RT PCR analysis, real time-QPCR, and histology. The results of several different analyses of the transplanted rMSCs embedded in the scaffolds showed that rMSCs coupled with a CS-bound TGF-beta 3 encapsulated scaffold evidenced superior cartilage tissue formation as measured by an assay of specific gene and protein expression. Moreover, the scaffold exhibited more rapid and more distinct morphology of differentiated rMSCs than was observed with other scaffolds, as determined by histology and immunochemical histology analysis. These results indicate that the elastic block copolymer scaffolds combined with a CS-bound TGF beta 3 should prove very suitable matrix for cell-based cartilage tissue engineering. PMID- 19280637 TI - Can low frequency electromagnetic field help cartilage tissue engineering? AB - To understand whether a low-frequency pulsed electromagnetic field (EMF) could help cartilage tissue repair in the scope of tissue engineering, we tested how EMF affected collagen gel properties and the behaviors of chondrocyte cells embedded in collagen constructs. Collagen gel and primary chondrocytes embedded in collagen were exposed to EMF for 24 h. Gel and cells that were not exposed to EMF served as controls. Collagen gel exposed to EMF was more hydrophobic and less susceptible to enzymatic degradation (both p < 0.05) than control. Three weeks after EMF exposure, chondrocytes showed higher proliferation and lower glycosaminoglycan (GAG) production (both p < 0.05) than control. By the end of the third week, aggrecan, type I, II, and X collagen mRNA expressions in the EMF group were 1.8 times higher (p < 0.05), except for type II collagen) than control. The increase in gene expression did not show up in aggrecan histological staining and type II and type X collagen immunohistochemical staining. Cells from both groups kept a normal polygonal shape through out the test period. Our results suggested that one-time EMF exposure could promote collagen-embedded chondrocytes proliferation and their gene expressions. It also promoted short term (week 1) GAG production and lacuna formation. No apparent GAG and type II collagen production was seen in histological staining three weeks after the EMF exposure. PMID- 19280638 TI - Dermis-derived hydrogels support adipogenesis in vivo. AB - Biomaterials that support adipogenesis could contribute to tissue engineering therapies to be used as alternatives to traditional methods of tissue reconstruction and regeneration. We have recently shown that hydrogels comprised of urea soluble proteins and polysaccharides extracted from adipose tissue promote preadipocyte differentiation in vitro and adipogenesis in vivo. However, it is not clear if these findings result from the adipose tissue source of the extracts or if the technique isolates adipogenic factors from other tissues. The present study investigates whether the application of this technique to dermis samples would provide adipogenic hydrogels. Extracts from dermis assembled into hydrogels by either temperature or pH mechanisms. Both formulations supported preadipocyte differentiation in vitro and vascularized adipose formation in vivo. The temperature formulation of the gels induced more rapid adipose formation than the pH formulation in vivo. Interestingly, in comparison to our previous studies the dermis derived hydrogels had comparable adipogenic properties to adipose gels in vivo but not in vitro. Further study of these materials could lead to insight of the role of specific matrix properties on adipogenesis. PMID- 19280639 TI - Toward an antitumor form of bovine pancreatic ribonuclease: the crystal structure of three noncovalent dimeric mutants. AB - The cytotoxic action of bovine seminal ribonuclease (BS-RNase) depends on its noncovalent swapped dimeric form (NCD-BS), which presents a compact structure that allows the molecule to escape ribonuclease inhibitor (RI). A key role in the acquisition of this structure has been attributed to the concomitant presence of a proline in position 19 and a leucine in position 28. The introduction of Leu28, Cys31, and Cys32 and, in addition, of Pro19 in the sequence of bovine pancreatic ribonuclease (RNase A) has produced two dimeric variants LCC and PLCC, which do exhibit a cytotoxic activity, though at a much lower level than BS-RNase. The crystal structure analysis of the noncovalent swapped form (NCD) of LCC and PLCC, complexed with the substrate analogue 2 '-deoxycytidylyl(3 ',5 ')-2 ' deoxyguanosine, has revealed that, differently from NCD-BS, the dimers adopt an opened quaternary structure, with the two Leu residues fully exposed to the solvent, that does not hinder the binding of RI. Similar results have been obtained for a third mutant of the pancreatic enzyme, engineered with the hinge peptide sequence of the seminal enzyme (residues 16-22) and the two cysteines in position 31 and 32, but lacking the hydrophobic Leu residue in position 28. The comparison of these three structures with those previously reported for other ribonuclease swapped dimers strongly suggests that, in addition to Pro19 and Leu28, the presence of a glycine at the N-terminal end of the hinge peptide is also important to push the swapped form of RNase A dimer into the compact quaternary organization observed for NCD-BS. PMID- 19280640 TI - Keratin-lipid structural organization in the corneous layer of snake. AB - The shed epidermis (molt) of snakes comprises four distinct layers. The upper two layers, here considered as beta-layer, contain essentially beta-keratin. The following layer, known as mesos-layer, is similar to the human stratum corneum, and is formed by thin cells surrounded by intercellular lipids. The latter layer mainly contains alpha-keratin. In this study, the molecular assemblies of proteins and lipids contained in these layers have been analyzed in the scale of two species of snakes, the elapid Tiger snake (TS, Notechis scutatus) and the viperid Gabon viper (GV, Bitis gabonica). Scanning X-ray micro-diffraction, FTIR and Raman spectroscopies, thermal analysis, and scanning electron microscopy experiments confirm the presence of the three layers in the GV skin scale. Conversely, in the TS molt a typical alpha-keratin layer appears to be absent. In the latter, experimental data suggest the presence of two domains similar to those found in the lipid intercellular matrix of stratum corneum. X-ray diffraction data also allow to determine the relative orientation of keratins and lipids. The keratin fibrils are randomly oriented inside the layers parallel to the surface of scales while the lipids are organized in lamellar structures having aliphatic chains normal to the scale surface. The high ordered lipid organization in the mature mesos layer probably increases its effectiveness in limiting water-loss. PMID- 19280641 TI - Mg2+ ions bind at the C-terminal region of skeletal muscle alpha-tropomyosin. AB - Tropomyosin (Tm) is a dimeric coiled-coil protein that polymerizes through head to-tail interactions. These polymers bind along actin filaments and play an important role in the regulation of muscle contraction. Analysis of its primary structure shows that Tm is rich in acidic residues, which are clustered along the molecule and may form sites for divalent cation binding. In a previous study, we showed that the Mg(2+)-induced increase in stability of the C-terminal half of Tm is sensitive to mutations near the C-terminus. In the present report, we study the interaction between Mg(2+) and full-length Tm and smaller fragments corresponding to the last 65 and 26 Tm residues. Although the smaller Tm peptide (Tm(259-284(W269))) is flexible and to large extent unstructured, the larger Tm(220-284(W269)) fragment forms a coiled coil in solution whose stability increases significantly in the presence of Mg(2+). NMR analysis shows that Mg(2+) induces chemical shift perturbations in both Tm(220-284(W269)) and Tm(259 284(W269)) in the vicinity of His276, in which are located several negatively charged residues. PMID- 19280642 TI - Exploring the interactions between signal sequences and E. coli SRP by two distinct and complementary crosslinking methods. AB - Photoaffinity crosslinking comprises a group of invaluable techniques used to investigate in detail a binding interaction between two polypeptides. As the diverse photo crosslinking techniques available display inherent differences, the method of choice will provide specific information about a particular system under study. We used two complementary crosslinking approaches: photo-induced crosslinking of unmodified proteins (PICUP) and benzophenone-mediated (BPM) crosslinking to extensively examine the interaction between the signal recognition particle (SRP) and signal sequences. Signal peptide binding by SRP presents a central puzzle in the protein targeting process because signal sequences must be recognized with fidelity but lack strict primary structural homology. The concurrent use of PICUP and BPM crosslinking to link signal peptides to E. coli SRP allowed us to explore the crosslinking pattern resulting from using different crosslinking chemistries, varying the position of the photoprobe in the hydrophobic core of the signal sequence, and shifting the crosslinking reactive group away from the signal peptide backbone. By PICUP, signal peptides crosslinked exclusively to the NG domain of the SRP protein Ffh, regardless of the position of the reactive residue. Benzophenone-modified amino acids preferentially crosslinked the signal peptide to the C-terminal (M) domain of Ffh. We conclude that signal peptide binding is largely mediated by the M domain. Importantly, our data also indicate intimate, at least transient, contacts between the hydrophobic core of the signal peptide and the NG domain. These results reopen the possibility of a direct involvement of the NG domain in signal sequence recognition. PMID- 19280643 TI - Evaluation of self-expandable, FK506-coated, covered stents in canine animal model. AB - This study aimed to evaluate whether drug coating of the recently developed covered SENDAI stents--self-expandable stents covered with segmented polyurethane (SPU) films--reduces neointimal thickening in animal model. FK506, which is one of the most effective immunosuppressants, was used. Bare stents; non-coated, covered stents; and FK506-coated, covered stents were placed bilaterally in the external iliac arteries of beagle dogs. After 1-month observation period, angiography did not show significant stent-induced stenosis. Histological evaluation revealed a completely endothelialized intravascular lumen and the absence of thrombus formation. The area of the intimal thickening induced by the FK506-coated stents was significantly smaller than that induced by the non-coated stents, whereas it was larger in the case of both the covered stents than that in the case of the bare stent. In conclusion, FK506 treatment of the self expandable, covered stents was confirmed to effectively inhibit intimal thickening, although the SPU film used for covering functioned as a drug carrier in addition to a scaffold for intimal formation. PMID- 19280644 TI - NMP-modified PMMA bone cement with adapted mechanical and hardening properties for the use in cancellous bone augmentation. AB - The use of polymethylmethacrylate (PMMA) to reinforce vertebral bodies (Vertebroplasty) leads to an increase in the Young's modulus of the augmented vertebral body. Fractures in the adjacent vertebrae may be the consequence thereof. Hence, PMMA with a reduced Young's modulus may be suitable for vertebroplasty. The goal of this study was to produce and characterize stiffness adapted PMMA cements. Modified PMMA bone cements were produced by adding N-methyl pyrrolidone (NMP). Young's modulus, yield strength, polymerization temperature, setting time, and hardening behavior of different cements were analyzed. Focus was on the mechanical properties of the material after different storage conditions (in air at room temperature and in PBS at 37 degrees C). The Young's modulus decreased from 2670 MPa (air)/2384 MPa (PBS) for the regular cement to 76 MPa (air)/320 MPa (PBS) for a material composition with 60% of the MMA substituted by NMP. Yield strength decreased from 85 MPa (air)/78 MPa (PBS) to 2 MPa (air)/24 MPa (PBS) between the regular cement and the 60% composition. Polymerization temperature decreased from 70 degrees C (regular cement) to 48 degrees C for the 30% composition. The hardening behavior exhibited an extension in handling time up to 200% by the modification presented. Modification of PMMA cement using NMP seems to be a promising method to make the PMMA cement more compliant for the use in cancellous bone augmentation in osteoporotic patients: adjustment of its mechanical properties close to those of cancellous bone, lower polymerization temperature, and extended handling time. PMID- 19280645 TI - Antibacterial activity and bonding ability of an adhesive incorporating an antibacterial monomer DMAE-CB. AB - This study evaluated the antibacterial effect and microtensile bond strength of a resin-based adhesive containing an antibacterial monomer DMAE-CB (methacryloxylethyl cetyl dimethyl ammonium chloride). Cured specimens of 1, 2, and 3% DMAE-CB-containing Single Bond 2 (crosslinking monomer: Bis-GMA, dimethacrylates; functional monomer: HEMA) were prepared, and their antibacterial effects on Streptococcus mutans ATCC 25175 were investigated. Antibacterial property after 0, 30, 90, and 180 days of aging was also tested. Bonding ability of the experimental adhesive incorporating 3% DMAE-CB was evaluated by microtensile bond strength test. The cured experimental adhesive exhibited an inhibitory effect on S. mutans growth, and the adhesive containing 3% DMAE-CB showed higher antibacterial efficiency compared with those incorporating 1 or 2% anibacterial monomer. Antibacterial activities of the specimens lasted for at least 180 days. Microtensile bond strength test revealed that the bonding ability of the experimental adhesive was not significantly adversely affected by the incorporation of DMAE-CB. Therefore, dental adhesives with strong and long lasting bacteriostatic property could be achieved by incorporating DMAE-CB without negatively influencing bonding ability. PMID- 19280647 TI - The dependence on gp130 cytokines of axotomy induced neuropeptide expression in adult sympathetic neurons. AB - Adult peripheral neurons exhibit dramatic changes in gene expression after axonal injury, including changes in neuropeptide phenotype. For example, sympathetic neurons in the superior cervical ganglion (SCG) begin to express vasoactive intestinal peptide (VIP), galanin, pituitary adenylate cyclase activating polypeptide (PACAP), and cholecystokinin after axotomy. Before these changes, nonneuronal cells in the SCG begin to express leukemia inhibitory factor (LIF). When the effects of axotomy were compared in LIF-/- and wild-type mice, the increases in VIP and galanin expression were less in the former, though significant increases still occurred. LIF belongs to a family of cytokines with overlapping physiological effects and multimeric receptors containing the subunit gp130. Real-time PCR revealed large increases in the SCG after axotomy in mRNA for three members of this cytokine family, interleukin (IL)-6, IL-11, and LIF, with modest increases in oncostatin M, no changes in ciliary neurotrophic factor, and decreases in cardiotrophin-1. To explore the role of these cytokines, animals with selective elimination of the gp130 receptor in noradrenergic neurons were studied. No significant changes in mRNA levels for VIP, galanin, and PACAP were seen in axotomized ganglia from these mutant mice, while the increase in cholecystokinin was as large as that seen in wild-type mice. The data indicate that the inductions of VIP, galanin, and PACAP after axotomy are completely dependent on gp130 cytokines and that a second cytokine, in addition to LIF, is involved. The increase in cholecystokinin after axotomy, however, does not require the action of these cytokines. PMID- 19280649 TI - Assessing high-resolution melt curve analysis for accurate detection of gene variants in complex DNA fragments. AB - Mutation detection has, until recently, relied heavily on the use of gel-based methods that can be both time consuming and difficult to design. Nongel-based systems are therefore important to increase simplicity and improve turn around time without compromising assay sensitivity and accuracy, especially in the diagnostic/clinical setting. In this study, we assessed the latest of the nongel based methods, namely high-resolution melt (HRM) curve analysis. HRM is a closed tube method that incorporates a saturating dye during DNA amplification followed by a monitoring of the change in fluorescence as the DNA duplex is denatured by an increasing temperature. We assessed 10 amplicons derived from eight genes, namely SERPINA1, CXCR7, MBL, VDR, NKX3A, NPY, TP53, and HRAS using two platforms, the LightScanner System using LC Green PLUS DNA binding dye (Idaho Technology, Salt Lake City, UT, USA) and the LightCycler 480 using the HRM Master dye (Roche Diagnostics, Indianapolis, IN, USA). DNA variants (mutations or polymorphims) were previously identified using denaturing gradient gel electrophoresis (DGGE) a method, similarly to HRM, based upon the different melting properties of double stranded DNA. Fragments were selected based on variant and fragment complexity. This included the presence of multiple sequence variants, variants in alternate orientations, and single or multiple variants (constitutional or somatic) in GC rich fragments. We demonstrate current limitations of the HRM method for the analysis of complex DNA regions and call for caution when using HRM as the sole method to make a clinical diagnosis based on genetic analysis. PMID- 19280648 TI - Hippocampal lesions impair spatial memory performance, but not song--a developmental study of independent memory systems in the zebra finch. AB - Songbirds demonstrate song- and spatial-learning, forms of memory that appear distinct in formal characteristics and fitting the descriptions and criteria of procedural and episodic-like memory function, respectively. As in other vertebrates, the neural pathways underlying these forms of memory may also be dissociable, and include the corresponding song circuit and hippocampus (HP). Whether (or not) these two memory systems interact is unknown. Interestingly, the HP distinguishes itself as a site of immediate early gene expression in response to song and as a site of estrogen synthesis, a steroid involved in song learning. Thus, an interaction between these memory systems and their anatomical substrates appears reasonable to hypothesize, particularly during development. To test this idea, juvenile male or female zebra finches received chemical lesions of the HP at various points during song learning, as did adults. Song structure, singing behavior, song preference, and spatial memory were tested in adulthood. Although lesions of the HP severely compromised HP-dependent spatial memory function across all ages and in both sexes, we were unable to detect any effects of HP lesions on song learning, singing, or song structure in males. Interestingly, females lesioned as adults, but not as juveniles, did lose the characteristic preference for their father's song. Since compromise of the neural circuits that subserve episodic-like memory does very little (if anything) to affect procedural like (song learning) memory, we conclude that these memory systems and their anatomical substrates are well dissociated in the developing male zebra finch. PMID- 19280650 TI - Disease-causing mutations improving the branch site and polypyrimidine tract: pseudoexon activation of LINE-2 and antisense Alu lacking the poly(T)-tail. AB - Cryptic exons or pseudoexons are typically activated by point mutations that create GT or AG dinucleotides of new 5' or 3' splice sites in introns, often in repetitive elements. Here we describe two cases of tetrahydrobiopterin deficiency caused by mutations improving the branch point sequence and polypyrimidine tracts of repeat-containing pseudoexons in the PTS gene. In the first case, we demonstrate a novel pathway of antisense Alu exonization, resulting from an intronic deletion that removed the poly(T)-tail of antisense AluSq. The deletion brought a favorable branch point sequence within proximity of the pseudoexon 3' splice site and removed an upstream AG dinucleotide required for the 3' splice site repression on normal alleles. New Alu exons can thus arise in the absence of poly(T)-tails that facilitated inclusion of most transposed elements in mRNAs by serving as polypyrimidine tracts, highlighting extraordinary flexibility of Alu repeats in shaping intron-exon structure. In the other case, a PTS pseudoexon was activated by an A>T substitution 9 nt upstream of its 3' splice site in a LINE-2 sequence, providing the first example of a disease-causing exonization of the most ancient interspersed repeat. These observations expand the spectrum of mutational mechanisms that introduce repetitive sequences in mature transcripts and illustrate the importance of intronic mutations in alternative splicing and phenotypic variability of hereditary disorders. PMID- 19280651 TI - Alu-Alu recombination underlies the vast majority of large VHL germline deletions: Molecular characterization and genotype-phenotype correlations in VHL patients. AB - Von Hippel-Lindau disease (VHL) is an autosomal dominant cancer syndrome. Affected individuals are predisposed to multiple tumors, primarily of the central nervous system (CNS), eyes, adrenals, and kidneys. The VHL tumor suppressor gene on chromosome 3p26-25 is partially or completely deleted in 20 to 30% of families with VHL. We identified deletions ranging from 0.5 kb to 250 kb affecting part of or the entire VHL and flanking genes in 54 families. In 33 of the index patients, the breakpoints were precisely characterized by DNA sequencing. Of the 66 breakpoints, 90% were located in Alu elements, revealing Alu-mediated recombination as the major mechanism for large germline deletions of the VHL gene, which lies in a region of high Alu density. Interestingly, an AluYa5 element in VHL intron 2, the evolutionarily youngest Alu element and the only such element in the entire region, was found to be the most recombinogenic, involved in 7 out of the 33 deletions. In comparison to VHL patients in general, the 54 index cases and their affected relatives showed a higher occurrence of renal cell carcinomas (RCC) and of CNS hemangioblastomas. We not only noted the association of RCC with retention of the HSPC300 gene, but also observed a significant correlation between retention of HSPC300 and the development of retinal angiomas (AR). This study reveals that germline VHL deletions provide a particularly rich source for the study of Alu-mediated unequal crossover events, and provides evidence for a protective role of the loss of the actin-regulator gene HSPC300 for the development of both RCC and AR. PMID- 19280652 TI - The prevalence of cervical ribs in a London population. AB - Cervical ribs are an important cause of neurovascular compression at the thoracic outlet. Previous studies have shown the prevalence of cervical ribs to be between 0.05 and 3%, depending on the sex and race of the population studied. We examined 1,352 chest radiographs to determine the prevalence of cervical ribs in a London population of mixed sex and ethnicity. Our study found that the overall prevalence of cervical ribs was 0.74% with a higher rate in females compared with males (1.09 and 0.42%, respectively). Of the 10 individuals with a cervical rib, five were on the left, three were on the right and two were bilateral. The presence of elongated C7 transverse processes (transverse apophysomegaly) was also noted. We found a total of 30 elongated transverse processes with an overall prevalence of 2.21%. They were also more common in females (3.43%) than males (1.13%). PMID- 19280653 TI - Henry Gray's Anatomy. AB - Little is generally known of Henry Gray, the author of Gray's Anatomy, and even less of his colleague Henry Vandyke Carter, who played a vital role in the dissections and illustrations leading to the production of the first volume in 1859. This essay attempts to sketch briefly the salient, know aspects of these two men and their divergent careers. It traces succinctly the subsequent fate of the unique anatomy book that has influenced and instructed almost every student of medicine. PMID- 19280654 TI - Attitudes to anatomy dissection in an Irish medical school. AB - Many studies around the world have looked at the stresses placed on medical students by cadaveric dissection. Although these studies have linked the use of cadavers in medical teaching to stress, some investigations have suggested an association with severe psychological stress and even post-traumatic stress disorder. This study assessed the attitudes of medical and biomedical sciences students in an Irish medical school towards cadaveric dissection by recording, through a questionnaire, their perceptions and experience before initial exposure to dissection and subsequently examining their attitudes after the first dissection and after 9 weeks. Student attitudes towards the dissecting room remained consistently positive for the duration of the study with only a minority of respondents reporting negative symptoms. Pre-existing attitudes to the idea of dissection were unaffected by exposure and subsequent continuous experience of dissection. The majority of students in this study did not find the dissecting room experience stressful, and considered time spent in the dissecting room valuable. However, the proportion of students with negative experiences in the dissecting room was higher than has been reported in previous studies. Many respondents felt they could be better prepared for the dissecting room experience, indicating an increasing requirement for effective preparatory programmes. PMID- 19280655 TI - Anatomy of the posterior inferior cerebellar artery: relevance for C1-C2 puncture procedures. AB - Lateral C1-C2 puncture is associated with a number of complications including damage of aberrant posterior inferior cerebellar arteries (PICA). We propose to determine the frequency of cases in which the PICA originates or descends below C1, thus posing a risk to the patient undergoing lateral C1-C2 puncture. Two hundred and eleven consecutive patients who received bilateral or unilateral vertebral angiography for evaluation of cerebrovascular lesions were included in this study. In total, 346 PICAs were studied. Lateral vertebral angiograms were analyzed for the location of the origin of the PICA and the inferior-most portion of the PICA relative to three anatomical regions. Region 1 was defined as the region above the foramen magnum. Region 2 was defined as the region below the foramen magnum but above the inferior border of the posterior arch of C1. Region 3 was defined as the region below the inferior border of the posterior arch of C1. In 2 of 346 angiograms (0.6%) and 2 of 211 patients (0.9%), the caudal loop of the PICA descended to Region 3. The PICA did not originate in Region 3 in any cases. In approximately 1% of patients in our study, the caudal loop of the PICA descended into a region that may have placed it at risk for damage during C1-C2 puncture. Our study suggests that it may be important for radiologists to assess the arterial anatomy of the C1-C2 region before performing C1-C2 puncture. PMID- 19280656 TI - Distal biceps brachii tendon anatomy revisited from a surgical perspective. AB - The distal biceps brachii tendon is commonly susceptible to traumatic injury. This study aimed to describe the morphology of the distal biceps brachii tendon in relation to the commonly used endobutton repair of tendon rupture. The results suggested that the distal tendon is a series of distinct bands of variable number. These bands are obscured surgically by the tendon sheath. Upon opening this sheath, blunt dissection of the tendon released fibrous connections between the tendon bands. Adjacent bands were variably connected via small oblique bands. The separations between bands were continuous onto the radius. They were therefore considered as separate force-conducting units. This notion is of high relevance to endobutton repairs, as the sutures are typically only passed through the margins of the tendon. Where few connections exist between tendinous bands, this represents a potential weakness, as central bands are therefore free to be pulled proximally. This is of primary concern in the early rehabilitative stages of postoperative care. It may be suggested that sutures that cross the width of the tendon will eliminate the give of central bands, improving postoperative results, reducing revision numbers, and potentially reducing rehabilitation time. PMID- 19280657 TI - Detection of mosaic RB1 mutations in families with retinoblastoma. AB - The RB1 gene mutation detection rate in 1,020 retinoblastoma families was increased by the use of highly sensitive allele specific-PCR (AS-PCR) to detect low-level mosaicism for 11 recurrent RB1 CGA>TGA nonsense mutations. For bilaterally affected probands, AS-PCR increased the RB1 mutation detection sensitivity from 92.6% to 94.8%. Both RB1 oncogenic changes were detected in 92.7% of sporadic unilateral tumors (357/385); 14.6% (52/357) of unilateral probands with both tumor mutations identified carried one of the tumor mutations in blood. Mosaicism was evident in 5.5% of bilateral probands (23 of 421), in 3.8% of unilateral probands (22 of 572), and in one unaffected mother of a unilateral proband. Half of the mosaic mutations were only detectable by AS-PCR for the 11 recurrent CGA>TGA mutations, and not by standard sequencing. This suggests that significant numbers of low-level mosaics with other classes of RB1 mutations remain unidentified by current technology. We show that the use of linkage analysis in a two-generation retinoblastoma family resulted in the erroneous conclusion that a child carried the parental mutation, because the founder parent was mosaic for the RB1 mutation. Of 142 unaffected parental pairs tested, only one unaffected parent of a proband (0.7%) showed somatic mosaicism for the proband's mutation, in contrast to an overall 4.5% somatic mosaicism rate for retinoblastoma probands, suggesting that mosaicism for an RB1 mutation is highly likely to manifest as retinoblastoma. PMID- 19280658 TI - Comparison of B-scan sonographic measurements of optic cup with fundus photographic measurements. AB - PURPOSE: To compare the measurements of the optic cup diameter with B-scan sonography with fundus photography in patients with clear ocular media and to propose a solution for the clinical problem of determining the cup-disc ratio in eyes with opaque ocular media. METHODS: We studied 122 patients (glaucoma patients and non-glaucoma controls) with clear ocular media. Measurements of optic cup diameters using Quantel 10 MHz B-scan sonography were compared with magnification-corrected measurements of the same area using Zeiss FF450 Plus fundus camera. Paired-sample t-test was used to compare the measurements. The correlation between measurements of optic cup diameters using the 2 methods was established. In addition, the optic disc diameters of both eyes were measured on fundus photographs and compared with each other. RESULTS: There was no significant difference between the fundus photographic and sonographic measurements of optic cup diameters. There was no difference in the fundus photographic measurements of optic disc diameters between the 2 eyes. CONCLUSION: In an eye with opaque ocular media, the cup-disc ratio can be calculated by using the optic cup diameter of this eye with B-scan sonography and the optic disc diameter of the fellow eye with clear media measured by fundus photography. PMID- 19280659 TI - Biliary dilatation secondary to lithiasis in a child affected by Langerhans' cell histiocytosis. AB - Langerhans' cell histiocytosis (LCH) is a disease of unknown pathogenesis, caused by clonal proliferation of Langerhans' cells. Liver involvement results in a cholangiopathy, which has the radiologic appearance of sclerosing cholangitis. Only 1 case of obstructive jaundice due to common bile duct stone in a patient with LCH has been described. We present a case of a 31-month-old child with LCH and liver involvement on the waiting list for liver transplantation. During the follow-up, there was a rapid onset of jaundice due to sludge and lithiasis. The patient was treated first with an endoscopic biliary plastic stent and then with percutaneous biliary drainage and bilioenteric anastomosis. PMID- 19280660 TI - Genetic ablation of tenascin-C expression leads to abnormal hippocampal CA1 structure and electrical activity in vivo. AB - Despite evidence that the extracellular matrix glycoprotein tenascin-C (TNC) is implicated in brain development and plasticity, its roles in the intact adult brain are unknown. Here we report that spontaneous local field potential (LFP) activity in freely moving adult TNC-deficient mice is abnormal. The power of cortical and hippocampal theta and gamma oscillations was enhanced in comparison to wild-type mice. The alteration in hippocampal gamma rhythm was subfield specific, such that CA1 gamma was accentuated while dentate gyrus gamma was normal. Similar to LFP, synaptic transmission and plasticity at perforant path synapses in the dentate gyrus were unaffected by the mutation. Morphological analyses revealed a subfield-specific reduction in the CA1 volume and a reduction in the numbers of somatostatin-positive interneurons in the hippocampus as potential structural substrates of the observed functional aberrations. These findings indicate a role for tenascin-C in structural organization of the CA1 hippocampal subfield and in shaping neural activity. PMID- 19280661 TI - Voluntary exercise and caloric restriction enhance hippocampal dendritic spine density and BDNF levels in diabetic mice. AB - Diabetes may adversely affect cognitive function, but the underlying mechanisms are unknown. To investigate whether manipulations that enhance neurotrophin levels will also restore neuronal structure and function in diabetes, we examined the effects of wheel running and dietary energy restriction on hippocampal neuron morphology and brain-derived neurotrophic factor (BDNF) levels in db/db mice, a model of insulin resistant diabetes. Running wheel activity, caloric restriction, or the combination of the two treatments increased levels of BDNF in the hippocampus of db/db mice. Enhancement of hippocampal BDNF was accompanied by increases in dendritic spine density on the secondary and tertiary dendrites of dentate granule neurons. These studies suggest that diabetes exerts detrimental effects on hippocampal structure, and that this state can be attenuated by increasing energy expenditure and decreasing energy intake. PMID- 19280662 TI - Hippocampal, retrosplenial, and prefrontal hypoactivity in a model of diencephalic amnesia: Evidence towards an interdependent subcortical-cortical memory network. AB - The medial diencephalon is vital for memory, but it is not known why. The present study tested between the predictions of current hypotheses as to why this region is critical for memory. Lesions were made in the rat mammillothalamic tract, the only diencephalic structure consistently associated with amnesia in humans after ischemia. Decreased activity, as measured by immediate-early gene expression (c fos), was found in three key sites associated with memory function: the hippocampus, the prefrontal cortex, and the retrosplenial cortex. The specificity of these changes was confirmed by the qualitatively different patterns of immediately-early gene changes seen after amygdala lesions, e.g., hypoactivity in the hippocampus and retrosplenial cortex following mammillothalamic tract lesions but not following amygdala lesions. The mammillothalamic lesion results unify substrates linked to diencephalic and temporal lobe amnesia, and thereby support a new account of diencephalic amnesia that emphasizes multiple dysfunctions across hippocampal, retrosplenial, and prefrontal areas. This account suggests a role for the mammillary bodies that is independent of their hippocampal inputs. PMID- 19280663 TI - Buried anterolateral thigh flap for pharyngoesophageal reconstruction: our method for monitoring. AB - BACKGROUND: A noninvasive method for monitoring in cases of pharyngoesophageal defects reconstruction with the anterolateral thigh flap is presented. METHODS: Seventeen patients underwent reconstruction from 2005 to 2007. In 16 patients, the distal stump of the descending branch of lateral circumflex femoral artery (LCFA) was left on the skin surface covered with a transparent film dressing and monitoring was performed by direct observation of the pulsation. In the 17th patient, the pedicle emerged from the transverse branch of the LCFA and dissection of the distal stump for monitoring was impossible; therefore, he was excluded from the study. RESULTS: Cessation of the pulsation of the stump was noticed immediately postoperatively in 1 patient because of occlusion of the arterial anastomosis and in a second case 15 hours postoperatively due to a neck hematoma that caused collapse of the vein of the pedicle. Both flaps were salvaged. CONCLUSION: The proposed method of monitoring is simple, reliable, costless, and easily interpreted by the nursing staff. PMID- 19280664 TI - Impact of an integrated care program for patients with head and neck cancer on the quality of care. AB - BACKGROUND: The management of patients with head and neck cancer is complex, and implementation of an integrated care program might improve the quality of care. METHODS: A prospective before-after study was performed in 1 clinic for head and neck oncology on 311 adults with head and neck cancer to evaluate an integrated care program. RESULTS: Scores on the integrated care indicators showed that the implementation of the integrated care program led to relevant improvements, eg, waiting time for diagnostic procedures less than 10 days (improvement of 37%), support for stopping smoking (+37%), nutrition support (+44%), assessment of CT and MRI scans by a an expert radiologist (+23%), and number of patients in contact with the specialist nurses (+37%). The program had no relevant effects on the outcome indicators. CONCLUSION: An integrated care program can improve several aspects of the management of patients with head and neck cancer. PMID- 19280665 TI - Quality of life in patients with maxillectomy prostheses. AB - BACKGROUND: This cross-sectional study sought to determine patient quality of life and function after prosthetic rehabilitation for maxillary and palate defects following cancer resection. METHODS: Sixty-nine patients were identified, 42 participated in the study (61%). The Obturator Functioning Scale (OFS) and 4 general quality of life measures (Mental Health Inventory [MHI], Impact of Events Scale [IES], Illness Intrusiveness Ratings Scale [IIRS], and Centre for Epidemiologic Studies Depression Scale [CES-D]) were correlated with clinical parameters. RESULTS: Leakage when swallowing foods was the most frequently reported problem with the obturator (29%). Positive correlation was noted between the OFS and both the IES subscales (p < .01) and CES-D (p < .001). Difficulty with speech and eating was associated with increased avoidance of social situations. The surgical approach had a significant effect on the OFS, IES, and MHI subscales (p < .01). CONCLUSION: These results support the findings that good obturator function is associated with a better quality of life. PMID- 19280666 TI - COX-2 polymorphisms and the risk for head and neck cancer in white patients. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) is an enzyme involved in the synthesis of prostaglandins and thromboxanes, which are regulators of processes such as inflammation, cell proliferation, and angiogenesis, all relevant for cancer development. We investigated whether functional genetic polymorphisms in COX-2 may have a risk-modifying effect on head and neck carcinogenesis. METHODS: Blood from 431 white patients with oral, pharyngeal, or laryngeal carcinoma and 438 white healthy controls was investigated for the presence of 2 functional promoter region polymorphisms (-1195A-->G and -765G-->C) in COX-2. RESULTS: Logistic regression analysis did not show differences in COX-2 genotype distributions between patients and controls. Also no differences were found when stratified according to tumor localization, sex, or tobacco consumption. CONCLUSION: In contrast to earlier reports on the role of these COX-2 polymorphisms in mediating susceptibility to squamous esophageal carcinoma in a Chinese population, we could not demonstrate a risk-modifying effect in head and neck carcinogenesis in whites. PMID- 19280667 TI - Modeling a granule-based anaerobic ammonium oxidizing (ANAMMOX) process. AB - A mathematical model was developed to describe the anaerobic ammonium oxidation (ANAMMOX) process in a granular upflow anaerobic sludge blanket (UASB) reactor. ANAMMOX granules were cultivated in the UASB reactor by seeding aerobic granules. The granule-based reactor had a great N-loading resistant capacity. The model simulation results on the 1-year reactor performance matched the experimental data well. The yield coefficient for the growth and the decay rate coefficient of the ANAMMOX granules were estimated to be 0.164 g COD g(-1) N and 0.00016 h(-1), respectively. With this model, the effects of process parameters on the reactor performance were evaluated. Results showed that the optimum granule diameter for the maximum N-removal should be between 1.0 and 1.3 mm and that the optimum N loading rate should be 0.8 kg N m(-3) d(-1). In addition, the substrate micro profiles in the ANAMMOX granules were measured with a microelectrode to explore the diffusion dynamics within the granules, and the measured profiles matched the predicted results well. PMID- 19280668 TI - Development and characterization of a tissue engineered pancreatic substitute based on recombinant intestinal endocrine L-cells. AB - A tissue engineered pancreatic substitute (TEPS) consisting of insulin-producing cells appropriately designed and encapsulated to support cellular function and prevent interaction with the host may provide physiological blood glucose regulation for the treatment of insulin dependent diabetes (IDD). The performance of agarose-based constructs which contained either a single cell suspension of GLUTag-INS cells, a suspension of pre-aggregated GLUTag-INS spheroids, or GLUTag INS cells on small intestinal submucosa (SIS), was evaluated in vitro for total cell number, weekly glucose consumption and insulin secretion rates (GCR and ISR), and induced insulin secretion function. The three types of TEPS studied displayed similar number of cells, GCR, and ISR throughout 4 weeks of culture. However, the TEPS, which incorporated SIS as a substrate for the GLUTag-INS cells, was the only type of TEPS tested which was able to retain the induced insulin secretion function of non-encapsulated GLUTag-INS cells. Though improvements in the expression level of GLUTag-INS cells and/or the number of viable cells contained within the TEPS are needed for successful treatment of a murine model of IDD, this study has revealed a potential method for promoting proper cellular function of recombinant L-cells upon incorporation into an implantable three-dimensional TEPS. PMID- 19280669 TI - Scale-up analysis for a CHO cell culture process in large-scale bioreactors. AB - Bioprocess scale-up is a fundamental component of process development in the biotechnology industry. When scaling up a mammalian cell culture process, it is important to consider factors such as mixing time, oxygen transfer, and carbon dioxide removal. In this study, cell-free mixing studies were performed in production scale 5,000-L bioreactors to evaluate scale-up issues. Using the current bioreactor configuration, the 5,000-L bioreactor had a lower oxygen transfer coefficient, longer mixing time, and lower carbon dioxide removal rate than that was observed in bench scale 5- and 20-L bioreactors. The oxygen transfer threshold analysis indicates that the current 5,000-L configuration can only support a maximum viable cell density of 7 x 10(6) cells mL(-1). Moreover, experiments using a dual probe technique demonstrated that pH and dissolved oxygen gradients may exist in 5,000-L bioreactors using the current configuration. Empirical equations were developed to predict mixing time, oxygen transfer coefficient, and carbon dioxide removal rate under different mixing related engineering parameters in the 5,000-L bioreactors. These equations indicate that increasing bottom air sparging rate is more efficient than increasing power input in improving oxygen transfer and carbon dioxide removal. Furthermore, as the liquid volume increases in a production bioreactor operated in fed-batch mode, bulk mixing becomes a challenge. The mixing studies suggest that the engineering parameters related to bulk mixing and carbon dioxide removal in the 5,000-L bioreactors may need optimizing to mitigate the risk of different performance upon process scale-up. PMID- 19280670 TI - Test of the relationship between sutural ossicles and cultural cranial deformation: results from Hawikuh, New Mexico. AB - A number of researchers have hypothesized that the biomechanical forces associated with cultural cranial deformation can influence the formation of sutural ossicles. However, it is still difficult to make definitive conclusions about this relationship because the effects appear to be quite weak, and contradictory results have been obtained when specific sutures and deformation types are compared across studies. This research retests the hypothesis using a single archeological sample of lamdoidally deformed, occipitally deformed, and undeformed crania from Hawikuh, New Mexico (AD 1300-1680). Our results show no significant difference in either the prevalence or number of ossicles between deformed and undeformed crania, suggesting that the abnormal strains generated by cranial shape modification during infancy are not a factor in ossicle development for this population. One significant relationship was detected at the right lambdoid suture in crania with asymmetrical occipital deformation. Crania that were more deformed on the left side showed greater numbers of ossicles on the right side, but the effect was small. Furthermore, the relationship may well reflect a sampling error, due to the small number of crania with greater left side deformation and scorable right side lambdoid ossicles (n = 11). Although it is possible that forms of cranial deformation other than the posterior tabular types examined here may affect ossicle expression, our review of the literature suggests that the relationship in humans is complex and incompletely understood at this time. PMID- 19280671 TI - Duetting--a mechanism to strengthen pair bonds in a dispersed pair-living primate (Lepilemur edwardsi)? AB - Duetting is defined as an interactively organized pair display in which one pair partner coordinates its vocalizations in time with those of the other. It is widespread among tropical birds and cohesive pair-living primates, in which it is suggested to strengthen pair bonds. We know very little about the presence and function of duetting in dispersed pair-living mammals. We studied duetting behavior in a solitary foraging, but pair-sleeping, primate, the Milne Edwards' sportive lemur, in a dry deciduous forest of north-western Madagascar. We radio tracked six pairs throughout 1 year and recorded their sleeping sites and associations, home-range use, and vocal and behavioral interactions. Three different periods were covered (mating, pregnancy, and offspring care). Sleeping partners form long-term pair bonds, indicated by an almost exclusive pair specific usage of sleeping sites and home-ranges across periods. We explored three functional hypothesis of duetting: mate reunion, pair reunion, and joint territorial defense. Pairs regularly engaged in duet calling. Duetting increased significantly during the offspring care period. Duetting occurred significantly more often at feeding sites than at sleeping sites. Pair partners synchronized behavioral activities after duetting. The activity most often synchronized was locomotion. Pair partners played an equal role in duetting with no difference between sexes in starting or terminating duetting. Altogether, our results provide support for the hypothesis that in dispersed pair-living primates, duetting evolved as a mechanism to coordinate activities between pair partners dispersed in space, to strengthen pair bonds, and, perhaps, to limit infanticide and nutritional stress in lactating females. PMID- 19280672 TI - Stable isotopic evidence for diet at the Imperial Roman coastal site of Velia (1st and 2nd centuries AD) in Southern Italy. AB - Here we report on a stable isotope palaeodietary study of a Imperial Roman population interred near the port of Velia in Southern Italy during the 1st and 2nd centuries AD. Carbon and nitrogen stable isotope analyses were performed on collagen extracted from 117 adult humans as well as a range of fauna to reconstruct individual dietary histories. For the majority of individuals, we found that stable isotope data were consistent with a diet high in cereals, with relatively modest contributions of meat and only minor contributions of marine fish. However, substantial isotopic variation was found within the population, indicating that diets were not uniform. We suggest that a number of individuals, mainly but not exclusively males, had greater access to marine resources, especially high trophic level fish. However, the observed dietary variation did not correlate with burial type, number of grave goods, nor age at death. Also, individuals buried at the necropolis at Velia ate much less fish overall compared with the contemporaneous population from the necropolis of Portus at Isola Sacra, located on the coast close to Rome. Marine and riverine transport and commerce dominated the economy of Portus, and its people were in a position to supplement their own stocks of fish with imported goods in transit to Rome, whereas at Velia marine exploitation existed side-by-side with land-based economic activities. PMID- 19280673 TI - Brief communication: Rethinking the impact of the 1918 influenza pandemic on sex differentials in mortality. AB - This study will assess the general impact of the 1918 influenza on overall mortality and its impact on mortality attributable to pulmonary tuberculosis in a small-scale population. Using life table and decomposition methodologies, changes in mortality in Gibraltar used a scheme that identified a pre-epidemic period (1904-1917), the epidemic year (1918), and the post-epidemic period (1919-1927). Overall health in both sexes fell significantly in 1918 with a drop in life expectancy at birth, however, health quickly rebounded in the post-epidemic period. In the case of women, there was a significant increase in life expectancy at birth after the epidemic. The impact of influenza on the magnitude of sex differentials in the life expectancy at birth fell during epidemic year but returned to a level comparable to that of the pre-epidemic period. With respect to respiratory tuberculosis deaths, the immediate impact of influenza was restricted to only a significant increase in the rate among women (aged 15-54). In the post-epidemic period, tuberculosis mortality rates returned to the pre epidemic state in both sexes. The findings from Gibraltar stand in contrast opposition to results reported for experience in the United States during the 1918 flu. PMID- 19280674 TI - Sex differences in humeral bilateral asymmetry in two hunter-gatherer populations: California Amerinds and British Columbian Amerinds. AB - This study uses two prehistoric Amerindian populations of hunter-gatherer subsistence patterns to determine whether levels of sexual dimorphism in humeral bilateral cross-sectional asymmetry are related to sex-specific differences in activities among these populations. Results confirmed that males of the California Amerind population who engaged in the more unimanual activities of spear hunting and warfare were more asymmetrical than were their female counterparts who engaged in the more bimanual activities of grinding acorns. California Amerind males were also more asymmetrical than British Columbian Amerind males who rowed (using both arms) extensively. Sex differences within British Columbian Amerinds were not statistically significant, nor were female differences between populations. In general, levels of humeral asymmetry appear to be more dependent on sex and population-specific behaviors rather than broad subsistence patterns. PMID- 19280675 TI - The causes of porotic hyperostosis and cribra orbitalia: a reappraisal of the iron-deficiency-anemia hypothesis. AB - Porosities in the outer table of the cranial vault (porotic hyperostosis) and orbital roof (cribra orbitalia) are among the most frequent pathological lesions seen in ancient human skeletal collections. Since the 1950s, chronic iron deficiency anemia has been widely accepted as the probable cause of both conditions. Based on this proposed etiology, bioarchaeologists use the prevalence of these conditions to infer living conditions conducive to dietary iron deficiency, iron malabsorption, and iron loss from both diarrheal disease and intestinal parasites in earlier human populations. This iron-deficiency-anemia hypothesis is inconsistent with recent hematological research that shows iron deficiency per se cannot sustain the massive red blood cell production that causes the marrow expansion responsible for these lesions. Several lines of evidence suggest that the accelerated loss and compensatory over-production of red blood cells seen in hemolytic and megaloblastic anemias is the most likely proximate cause of porotic hyperostosis. Although cranial vault and orbital roof porosities are sometimes conflated under the term porotic hyperostosis, paleopathological and clinical evidence suggests they often have different etiologies. Reconsidering the etiology of these skeletal conditions has important implications for current interpretations of malnutrition and infectious disease in earlier human populations. PMID- 19280676 TI - A new high-resolution computed tomography (CT) segmentation method for trabecular bone architectural analysis. AB - In the last decade, high-resolution computed tomography (CT) and microcomputed tomography (micro-CT) have been increasingly used in anthropological studies and as a complement to traditional histological techniques. This is due in large part to the ability of CT techniques to nondestructively extract three-dimensional representations of bone structures. Despite prior studies employing CT techniques, no completely reliable method of bone segmentation has been established. Accurate preprocessing of digital data is crucial for measurement accuracy, especially when subtle structures such as trabecular bone are investigated. The research presented here is a new, reproducible, accurate, and fully automated computerized segmentation method for high-resolution CT datasets of fossil and recent cancellous bone: the Ray Casting Algorithm (RCA). We compare this technique with commonly used methods of image thresholding (i.e., the half maximum height protocol and the automatic, adaptive iterative thresholding procedure). While the quality of the input images is crucial for conventional image segmentation, the RCA method is robust regarding the signal to noise ratio, beam hardening, ring artifacts, and blurriness. Tests with data of extant and fossil material demonstrate the superior quality of RCA compared with conventional thresholding procedures, and emphasize the need for careful consideration of optimal CT scanning parameters. PMID- 19280677 TI - A revised radiocarbon date for a case of treponemal disease from Safed, Israel, from the 15th Century AD. PMID- 19280678 TI - Predictors of neuropsychiatric symptoms in nursing home patients: influence of gender and dementia severity. AB - INTRODUCTION: The aim of the study was to assess the influence of dementia severity and gender on neuropsychiatric symptoms in demented nursing home patients. METHODS: Neuropsychiatric symptoms were assessed in a large sample of 1319 Dutch nursing home patients using the Cohen-Mansfield Agitation Inventory (CMAI) and the Neuropsychiatric Inventory-Nursing Home version (NPI-NH). The influence of gender and severity of cognitive decline, assessed with the Global Deterioration Scale (GDS), was investigated using logistic regression analysis and subsequently corrected for possible confounders, such as age, duration of institutionalization and psychoactive medication use. RESULTS: While physically aggressive behaviour was more common in patients with very severe cognitive deterioration (GDS stage 7), disinhibition, irritability, physically non aggressive and verbally agitated behaviour were more common in patients in GDS stage 5 or 6. Physically aggressive behaviour was more common in men, whereas female patients demonstrated more verbally agitated behaviour. With respect to other neuropsychiatric symptoms, delusions and depression were also more common in patients in GDS stage 5 and 6, while prevalences of anxiety and apathy further increased in severely demented patients (GDS stage 7). Apathy was more prevalent in male patients, while depression and anxiety were more common in females. CONCLUSION: In nursing home patients, neuropsychiatric symptoms were associated with the severity of dementia, with most symptoms occurring in patients showing (moderately) severe cognitive decline. Only physical aggression, anxiety and apathy were more common in patients with very severe cognitive decline. Dementia severity and gender were important predictors of neuropsychiatric symptoms in this patient cohort. PMID- 19280679 TI - Verbal cued recall as a predictor of conversion to Alzheimer's disease in Mild Cognitive Impairment. AB - OBJECTIVE: This study was set up to investigate whether neuropsychological tests are able to predict conversion to AD among Mild Cognitive Impairment (MCI) patients. METHODS: At baseline the cognitive part of the Cambridge Examination for Mental Disorders of the Elderly (CAMCOG), the Mini Mental Status Examination (MMSE), the Geriatric Depression Scale (GDS), a Dutch variation of Rey's Auditory Verbal Learning Test, the Memory Impairment Screen plus (MISplus) and the Visual Association Test (VAT) were administered to 40 patients diagnosed with MCI. After 18 months, MCI-patients were reassessed and a follow-up diagnosis was established. Of those who were seen for follow-up (n = 31), seven fulfilled (NINCDS-ADRDA) criteria of probable AD, while 24 did not convert. RESULTS: A binary logistic regression analysis showed that the MISplus contributed most to the prediction of conversion (OR = 0.28, 95% CI 0.099-0.790). With a cut-off of 2 out of 6, a positive predictive value of 71.5%, a negative predictive value of 91.5% and an overall diagnostic accuracy of 87.0% were achieved. CONCLUSIONS: This prospective, longitudinal study shows that a score of 0 or 1 out of 6 on the MISplus may be a good indicator of future (within 18 months) progression to AD among MCI-patients. PMID- 19280680 TI - Use of physical restraints and antipsychotic medications in nursing homes: a cross-national study. AB - OBJECTIVES: This study compares inter- and intra-country differences in the prevalence of physical restraints and antipsychotic medications in nursing homes, and examines aggregated resident conditions and organizational characteristics correlated with these treatments. METHODS: Population-based, cross-sectional data were collected using a standardized Resident Assessment Instrument (RAI) from 14,504 long-term care facilities providing nursing home level services in five countries participating in the interRAI consortium, including Canada, Finland, Hong Kong (Special Administrative Region, China), Switzerland, and the United States. Facility-level prevalence rates of physical restraints and antipsychotic use were examined both between and within the study countries. RESULTS: The prevalence of physical restraint use varied more than five-fold across the study countries, from an average 6% in Switzerland, 9% in the US, 20% in Hong Kong, 28% in Finland, and over 31% in Canada. The prevalence of antipsychotic use ranged from 11% in Hong Kong, between 26-27% in Canada and the US, 34% in Switzerland, and nearly 38% in Finland. Within each country, substantial variations existed across facilities in both physical restraint and antipsychotic use rates. In all countries, neither facility case mix nor organizational characteristics were particularly predictive of the prevalence of either treatment. CONCLUSIONS: There exists large, unexplained variability in the prevalence of physical restraint and antipsychotic use in nursing home facilities both between and within countries. Since restraints and antipsychotics are associated with adverse outcomes, it is important to understand the idiosyncratic factors specific to each country that contribute to variation in use rates. PMID- 19280681 TI - Depression status as a reliable predictor of functional decline among Japanese community-dwelling older adults: a 12-year population-based prospective cohort study. AB - OBJECTIVE: This study aimed to examine a longitudinal relationship between depression status and functional decline among Japanese community-dwelling older adults, using a 12-year population-based, prospective cohort study design. METHOD: A total of 710 men and women, aged 65 years and over, participated in the study. Katz's Index and the Tokyo Metropolitan Institute of Gerontology Index of Competence were used to measure the functional capacities of basic activities of daily living (BADL) and higher-level competence, respectively. For the purpose of analysis, a decline in each subscale of functional capacity during the follow-up period were used as outcome variables; depression status assessed by the Japanese version of the 30-item Geriatric Depression Scale (GDS), with a cut-off of 11, was used as an independent variable; and age, gender, education level, history of chronic disease, hospitalization, smoking, physical activity, living alone, hearing problems, physical pain, dietary habits, and usual walking speed at baseline were used as covariates. RESULTS: Use of the multivariate Cox proportional hazards model adjusted for potential confounders showed that depression status was significantly and independently associated with BADL decline (risk ratio (RR) = 1.46, 95% confidence interval (CI): 1.13-1.89) and with higher-level competence decline (RR = 1.56, 95% CI: 1.18-2.04). CONCLUSION: Our study found an independent relationship between depression status and longitudinal change in functional capacity among community-dwelling older individuals, suggesting that depression status is a reliable predictor of functional decline (both of BADL decline and higher-level competence decline) in older adults. PMID- 19280682 TI - Separation with zwitterionic hydrophilic interaction liquid chromatography improves protein identification by matrix-assisted laser desorption/ionization based proteomic analysis. AB - Comprehensive proteomic analyses necessitate efficient separation of peptide mixtures for the subsequent identification of proteins by mass spectrometry (MS). However, digestion of proteins extracted from cells and tissues often yields complex peptide mixtures that confound direct comprehensive MS analysis. This study investigated a zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) technique for the peptide separation step, which was verified by subsequent MS analysis. Human serum albumin (HSA) was the model protein used for this analysis. HSA was digested with trypsin and resolved by ZIC-HILIC or conventional strong cation exchange (SCX) prior to MS analysis for peptide identification. Separation with ZIC-HILIC significantly improved the identification of HSA peptides over SCX chromatography. Detailed analyses of the identified peptides revealed that the ZIC-HILIC has better peptide fractionation ability. We further demonstrated that ZIC-HILIC is useful for quantitatively surveying cell surface markers specifically expressed in undifferentiated embryonic stem cells. These results suggested the value of ZIC-HILIC as a novel and efficient separation method for comprehensive and quantitative proteomic analyses. PMID- 19280683 TI - Direct detection of alkylphosphonic acids in environmental matrices by proton coupled phosphorus NMR. AB - A simple, convenient, and direct one-dimensional (1D) (31)P NMR technique is demonstrated for the detection of alkylphosphonic acids (marker of nerve agents). The results of detection were validated after conducting various in-house exercises. The confidence generated by this study was found very useful in detection of different alkylphosphonic acids spiked in various official interlaboratory proficiency tests conducted by Organisation for the Prohibition of Chemical Weapons (OPCW). PMID- 19280684 TI - Is hepatic resection justified after chemotherapy in patients with colorectal liver metastases and lymph node involvement? PMID- 19280687 TI - Passive competition between pairs of self-ordering proto-replicators and the effect of primitive membranes. AB - Self-ordering proto-replicators provide an abstract, simulation based investigation of a possible intermediary step in the development of informational replicators. They involve the dynamic self-ordering of generalised subunits of at least two types into coherent and mobile structures, and do not require the preexistence of templates, replicators or functional catalysts. The following discussion naturally extends this approach to include pairs of such systems linked via a probabilistic, inter-system movement of structures. Initially for two specific forms of self-ordering proto-replicator, various types of probabilistic movement were studied, both with and without a simple model of a primitive membrane. Then for a specific probabilistic movement, which was again considered with and without membranes, different pairings of twelve forms of self ordering proto-replicators were studied. The results show that the systems exhibited a form of passive competition for subunits and that this produced a new level of multi-system self-ordering that depended upon the relative self-ordering capabilities of the individual systems. This effect was also found to be significantly more pronounced for those systems that included the primitive membranes. The implications for the origins of life were then considered. PMID- 19280686 TI - Alzheimer's disease neuroimaging initiative: a one-year follow up study using tensor-based morphometry correlating degenerative rates, biomarkers and cognition. AB - Tensor-based morphometry can recover three-dimensional longitudinal brain changes over time by nonlinearly registering baseline to follow-up MRI scans of the same subject. Here, we compared the anatomical distribution of longitudinal brain structural changes, over 12 months, using a subset of the ADNI dataset consisting of 20 patients with Alzheimer's disease (AD), 40 healthy elderly controls, and 40 individuals with mild cognitive impairment (MCI). Each individual longitudinal change map (Jacobian map) was created using an unbiased registration technique, and spatially normalized to a geometrically-centered average image based on healthy controls. Voxelwise statistical analyses revealed regional differences in atrophy rates, and these differences were correlated with clinical measures and biomarkers. Consistent with prior studies, we detected widespread cerebral atrophy in AD, and a more restricted atrophic pattern in MCI. In MCI, temporal lobe atrophy rates were correlated with changes in mini-mental state exam (MMSE) scores, clinical dementia rating (CDR), and logical/verbal learning memory scores. In AD, temporal atrophy rates were correlated with several biomarker indices, including a higher CSF level of p-tau protein, and a greater CSF tau/beta amyloid 1-42 (ABeta42) ratio. Temporal lobe atrophy was significantly faster in MCI subjects who converted to AD than in non-converters. Serial MRI scans can therefore be analyzed with nonlinear image registration to relate ongoing neurodegeneration to a variety of pathological biomarkers, cognitive changes, and conversion from MCI to AD, tracking disease progression in 3 dimensional detail. PMID- 19280688 TI - Regional shape abnormalities in mild cognitive impairment and Alzheimer's disease. AB - Magnetic resonance (MR) based shape analysis provides an opportunity to detect regional specificity of volumetric changes that may distinguish mild cognitive impairment (MCI) and Alzheimer's disease (AD) from healthy elderly controls (CON), and predict future conversion to AD. We assessed the surface deformation of seven structures (amygdala, hippocampus, thalamus, caudate, putamen, globus pallidus, body and temporal horn of the lateral ventricles) in 383 MRI volumes, based on data shared through the publicly available Alzheimer's Disease Neuroimaging Initiative (ADNI), to identify regionally-specific shape abnormalities in MCI and AD. Large deformation diffeomorphic metric mapping (LDDMM) was used to generate the shapes of seven structures based on template shapes injected into segmented subcortical volumes. LDDMM then constructed the surface deformation maps encoding the local shape variation of each subject relative to the template. Hierarchical models were developed to detect differences in local shape in MCI and AD relative to CON. Our findings revealed that surface inward-deformation in MCI and AD is most prominent in the anterior hippocampal segment and the basolateral complex of the amygdala. Most pronounced surface outward-deformation in MCI and AD occurs in the lateral ventricles. Mild surface inward-deformation in MCI and AD occurs in the anterior-lateral and ventral-lateral aspects of the thalamus, with no evidence of regionally-specific deformation in the putamen or globus pallidus. Although the locations of the shape abnormalities in MCI and AD are primarily within the mesial temporal region, analyses support distinct components of correlated shape variation that may help predict future MCI conversion. PMID- 19280689 TI - Nox4 overexpression activates reactive oxygen species and p38 MAPK in human endothelial cells. AB - Nicotine adenine dinucleotide phosphate (NADPH) oxidase (Nox) complexes are the main sources of reactive oxygen species (ROS) formation in the vessel wall. We have used DNA microarray, real-time PCR and Western blot to demonstrate that the subunit Nox4 is the major Nox isoform in primary human endothelial cells; we also found high levels of NADPH oxidase subunit p22(phox) expression. Nox4 was localized by laser scanning confocal microscopy within the cytoplasm of endothelial cells. Endothelial Nox4 overexpression enhanced superoxide anion formation and phosphorylation of p38 MAPK. Nox4 down-regulation by shRNA has in contrast to TGF-beta no effect on p38 MAPK phosphorylation. We conclude that Nox4 is the major Nox isoform in human endothelial cells, and forms an active complex with p22(phox). The Nox4-containing complex mediates formation of reactive oxygen species and p38 MAPK activation. This is a novel mechanism of redox-sensitive signaling in human endothelial cells. PMID- 19280690 TI - Campylobacter jejuni fatty acid synthase II: structural and functional analysis of beta-hydroxyacyl-ACP dehydratase (FabZ). AB - Fatty acid biosynthesis is crucial for all living cells. In contrast to higher organisms, bacteria use a type II fatty acid synthase (FAS II) composed of a series of individual proteins, making FAS II enzymes excellent targets for antibiotics discovery. The beta-hydroxyacyl-ACP dehydratase (FabZ) catalyzes an essential step in the FAS II pathway. Here, we report the structure of Campylobacter jejuni FabZ (CjFabZ), showing a hexamer both in crystals and solution, with each protomer adopting the characteristic hot dog fold. Together with biochemical analysis of CjFabZ, we define the first functional FAS II enzyme from this pathogen, and provide a framework for investigation on roles of FAS II in C. jejuni virulence. PMID- 19280691 TI - The underlying anatomical correlates of long-term meditation: larger hippocampal and frontal volumes of gray matter. AB - Although the systematic study of meditation is still in its infancy, research has provided evidence for meditation-induced improvements in psychological and physiological well-being. Moreover, meditation practice has been shown not only to benefit higher-order cognitive functions but also to alter brain activity. Nevertheless, little is known about possible links to brain structure. Using high resolution MRI data of 44 subjects, we set out to examine the underlying anatomical correlates of long-term meditation with different regional specificity (i.e., global, regional, and local). For this purpose, we applied voxel-based morphometry in association with a recently validated automated parcellation approach. We detected significantly larger gray matter volumes in meditators in the right orbito-frontal cortex (as well as in the right thalamus and left inferior temporal gyrus when co-varying for age and/or lowering applied statistical thresholds). In addition, meditators showed significantly larger volumes of the right hippocampus. Both orbito-frontal and hippocampal regions have been implicated in emotional regulation and response control. Thus, larger volumes in these regions might account for meditators' singular abilities and habits to cultivate positive emotions, retain emotional stability, and engage in mindful behavior. We further suggest that these regional alterations in brain structures constitute part of the underlying neurological correlate of long-term meditation independent of a specific style and practice. Future longitudinal analyses are necessary to establish the presence and direction of a causal link between meditation practice and brain anatomy. PMID- 19280692 TI - Collagen type V modulates fibroblast behavior dependent on substrate stiffness. AB - Collagen type V is highly expressed during tissue development and wound repair, but its exact function remains unclear. Cell binding to collagen V affects various basic cell functions and increased collagen V levels alter the structural organization and the stiffness of the ECM. We studied the combined effects of collagen V and substrate stiffness on the morphology, focal adhesion formation, and actin organization of fibroblasts. We found that a hybrid collagen I/V coating impairs fibroblast spreading on soft substrates (<10 kPa), but not on stiffer substrates (68 kPa or glass). In sharp contrast, a pure collagen I coating does not impair cell spreading on soft substrates. The impairment of cell spreading by collagen V is accompanied by diffuse actin staining patterns and small focal adhesions. These observations suggest that collagen V plays an essential role in modifying cell behavior during development and remodeling, when very soft tissues are present. PMID- 19280693 TI - Mapping functional connectivity based on synchronized CMRO2 fluctuations during the resting state. AB - Synchronized low-frequency fluctuations in the resting state functional MRI (fMRI) signal have been suggested to be associated with functional connectivity in brain networks. However, the underlying mechanism of this connectivity is still poorly understood, with the synchronized fluctuations could either originate from hemodynamic oscillations or represent true neuronal signaling. To better interpret the resting signal, in the current work, we examined spontaneous fluctuations at the level of cerebral metabolic rate of oxygenation (CMRO2), an index reflecting regional oxygen consumption and metabolism, and thus less sensitive to vascular dynamics. The CMRO2 signal was obtained based on a biophysical model with data acquired from simultaneous blood oxygenation level dependent (BOLD) and perfusion signals. CMRO2-based functional connectivity maps were generated in three brain networks: visual, default-mode, and hippocampus. Experiments were performed on twelve healthy participants during 'resting state' and as a comparison, with a visual task. CMRO2 signals in each of the above mentioned brain networks showed significant correlations. Functional connectivity maps from the CMRO2 signal are, in general, similar to those from BOLD and perfusion. In addition, we demonstrated that the three parameters (M, alpha and beta) in the biophysical model for calculating CMRO2 have negligible effects on the determination of the CMRO2-based connectivity strength. This study provides evidence that the spontaneous fluctuations in fMRI at rest likely originate from dynamic changes of cerebral metabolism reflecting neuronal activity. PMID- 19280694 TI - Modelling and analysis of time-variant directed interrelations between brain regions based on BOLD-signals. AB - Time-variant Granger Causality Index (tvGCI) was applied to simulated and measured BOLD signals to investigate the reliability of time-variant analysis approaches for the identification of directed interrelations between brain areas on the basis of fMRI data. Single-shot fMRI data of a single image slice with short repetition times (200 ms, 16000 frames/subject, 64x64 voxels) were acquired from 5 healthy subjects during an externally-driven, self-paced finger-tapping paradigm (57-59 single taps for each subject). BOLD signals were derived from the pre-supplementary motor area (preSMA), the supplementary motor area (SMA), and the primary motor cortex (M1). The simulations were carried out by means of a Dynamic Causal Modelling (DCM) approach. The tvGCI as well as time-variant Partial Directed Coherence (tvPDC) were used to identify the modelled connectivity network (connectivity structure - CS - of the DCM). Different CSs were applied by using dynamic systems (Generalized Dynamic Neural Network - GDNN) and trivariate autoregressive (AR) processes. The influence of the low-pass characteristics of the simulated hemodynamic response (Balloon model) and of the measuring noise was tested. Additionally, our modelling strategy considered "spontaneous" BOLD fluctuations before, during, and after the appearance of the event-related BOLD component. Couplings which were extracted from the simulated signals were statistically evaluated (tvGCI for shuffled data, confidence tubes for tvGCI courses). We demonstrate that connections of our CS models can be correctly identified during the event-related BOLD component and with signal-to noise-ratios corresponding to those of the measured data. The results based on simulations can be used to examine the reliability of connectivity identification based on BOLD signals by means of time-variant as well as time-invariant connectivity measures and enable a better interpretation of the analysis results using fMRI data. A readiness-BOLD response was only detected in one subject. However, in two subjects a strong time-variant connection (tvGCI) from preSMA to SMA was observed 3 s before the tapping was executed. This connection was accompanied by a weaker rise of the tvGCI from preSMA to M1. These preceding interrelations were confirmed in the other subjects by the dynamics of tvGCI courses. Based on the results of tvGCI analysis, the time-evolution of an individual connectivity network is shown for each subject. PMID- 19280695 TI - Genome-wide screening and characterization of transposable elements and their distribution analysis in the silkworm, Bombyx mori. AB - To elucidate the contribution of transposable elements (TEs) to the silkworm genome structure and evolution, we have conducted genome-wide analysis of TEs using the newly released genome assembly. The TEs made up 35% of the genome and contributed greatly to the genome size. Non-long terminal repeat retrotransposons (non-LTRs) and short interspersed nuclear elements (SINEs) were the predominant TE classes. From characterization of the TE distribution in the genome, it was revealed that non-LTRs, especially R1 clade elements, are frequently inserted into GC-rich regions. The GC content of non-LTRs themselves was over 40%, which indicate their contribution to the GC content of the insertion region. TEs accumulated in regions with low gene density, and there were relatively strong positive correlations between TE density and chromosomal recombination rate. We also characterized the clade distribution of the non-LTRs. The silkworm non-LTRs represented 10 of the 16 previously defined clades, which had the most variety than that reported for other genomes. Two partial CRE clade elements were found, which is one of the most ancient lineages of non-LTRs, and have been only found in Trypanosoma and fungi before. This analysis suggests that Bombyx genome is influenced by numerous amounts and variety of TEs. PMID- 19280696 TI - Chronic effects of flunitrazepam on shock-induced behavioral inhibition in adult male rats. AB - Flunitrazepam has been reported to trigger cases of extreme violence in man and recently it was shown to heighten aggression in experimental rats. The underlying mechanisms may involve disinhibited/impulsive behavior. Here, tolerance developed to the behavioral disinhibiting effect of flunitrazepam in Vogel's drinking conflict model, both in rats treated with flunitrazepam (0.32 mg/kg/day) for three weeks, as well as in rats treated for six weeks (0.32 mg/kg/day three weeks + 0.56 mg/kg/day three weeks). The results suggest that behavioral disinhibition is of limited importance in the mechanisms underlying the pro-aggressive effect of chronic flunitrazepam administration. PMID- 19280697 TI - Prochlorperazine--treatment for acute confusional migraine. AB - Acute confusional migraine is a rare migraine variant primarily seen in childhood that lacks standardized diagnostic criteria. Acute symptomatic treatment for this disorder has not been established. We report 2 patients having a total of 6 episodes of acute confusional migraine where the symptoms resolved with prochlorperazine with variable success. Intravenous prochlorperazine was highly effective in 3 out of 4 episodes. PMID- 19280698 TI - Memantine in the preventive treatment for migraine and refractory migraine. PMID- 19280699 TI - Genome-wide survey for baculoviral host homologs using the Bombyx genome sequence. AB - The Baculoviridae is a large family of pathogens that are infectious for arthropods, particularly insects of the Lepidoptera. Nucleopolyhedroviruses (NPVs), a genus of Baculoviridae, have a large circular, supercoiled, and double stranded DNA genome packaged into rod-shaped virions. The Bombyx mori NPV (BmNPV), an NPV pathogenic for B. mori, is known to potentially encode 136 proteins. Using the B. mori genome information, we found that 15 of 136 BmNPV proteins (11%) show significant similarity to the B. mori proteins. Among them, genes encoding nine proteins can be deleted in B. mori cultured cell line BmN by homologous recombination, indicating that these genes are dispensable for normal virus production. Interestingly, most of non-essential auxiliary genes encode proteins controlling host physiology at cellular and/or organismal levels: ecdysteroid UDP-glucosyltransferase inactivates an insect molting hormone ecdysone, protein tyrosine phosphatase is involved in wandering behavior at the late stage of infection, fibroblast growth factor induces host cell chemotaxis, and chitinase and cathepsin are required for postmortem host liquefaction. Deletion analysis of other non-essential genes also showed that three of them are viral pathogenicity factors for B. mori. These findings suggest that the modern lepidopteran baculovirus may have acquired auxiliary genes from an ancestral host insect to control host physiology and to increase the efficiency of virus transmission in nature. PMID- 19280700 TI - A comparison of morphometric techniques for studying the shape of the corpus callosum in adolescent idiopathic scoliosis. AB - The purpose of this paper is to compare volume- and boundary-based morphometry methods by applying them in the statistical analysis of 2-D shapes. The methods discussed in the first category include voxel-, deformation-, and tensor-based morphometry. The active shape model is demonstrated as an example of the second category of methods. The test data are 2-D shapes of the corpus callosum (CC) obtained in patients with left-thoracic adolescent idiopathic scoliosis (AIS), as well as age and sex matched healthy participants. The features of both categories of methods and the complementarily of them are demonstrated, which may provide guidelines for their applications in medical-image analysis. The morphometric abnormality in the splenium of the CC cross-validated by different methods has potential value in the prognosis and curve prediction of the left-thoracic AIS. PMID- 19280701 TI - A genomewide survey of homeobox genes and identification of novel structure of the Hox cluster in the silkworm, Bombyx mori. AB - Homeobox genes encode transcriptional factors that play crucial roles in a variety of developmental pathways from unicellular to multicellular eukaryotes. We have identified 102 homeobox genes in the typical insect of Lepidoptera, Bombyx mori, based on the newly assembled genome sequence with 9X coverage. These identified homeobox genes were categorized into nine classes including at least 74 families. The available ESTs and microarray data at present confirmed that more than half of them were expressed during silkworm developmental processes. Orthologs of pb, zen and ftz were newly identified in the Bombyx Hox cluster on chromosome 6. Interestingly, a special group of 12 tandemly duplicated homeobox genes was found located between Bmpb and Bmzen in the Bombyx Hox cluster, suggesting that Hox cluster might have experienced a lineage-specific expansion in the silkworm. A detailed analysis on genome data reveals that a split exists between Bmlab and Bmpb. Our data provide valuable information for future research on the development and evolution of silkworm. PMID- 19280702 TI - Salivary DNA, lipid, and protein oxidation in nonsmokers with periodontal disease. AB - Reactive oxygen species (ROS) are implicated in the destruction of the periodontium during periodontitis. The imbalance in oxidant activity may be a key factor. The aim of this paper is to determine whether periodontitis is associated with increased oxidative damage to DNA, lipids, and proteins and modification of total antioxidant capacity (TAC) in saliva. Saliva was collected from 58 periodontitis patients and 234 healthy controls, all nonsmokers. Periodontal disease status was characterized using the Community Periodontal Index of Treatment Needs (CPITN). Assays for 8-OHdG (ELISA), 8-epi-PGF2alpha (ELISA), and total protein carbonyls (ELISA), and oxy-blotting (Western)/mass spectrometry were performed to quantify oxidative damage to nucleic acids, lipids, total and individual proteins, respectively, in whole nonstimulated saliva. Salivary TAC was measured by inhibition of ABTS oxidation by metmyoglobin. We observed (i) significantly higher levels of 8-OHdG, 8-epi-PGF2alpha, and carbonylated proteins in saliva of periodontal patients as compared with controls (P=0.0003, <0.0001 and <0.0001); (ii) 8-OHdG, 8-epi-PGF2alpha, and carbonylated proteins were independently negatively associated with CPITN (P=0.004, 0.02, and <0.0001); (iii) a positive correlation between salivary TAC and periodontal disease status in the study group (P<0.0001); and (iv) specific oxidation of transferrin, human IgG1 heavy chain fragment, and salivary amylase in periodontitis. Periodontal disease is associated with increased oxidative modification of salivary DNA, lipids, and proteins. Augmented salivary total antioxidant capacity may represent an adaptive response to oxidative stress. Salivary amylase, transferrin, and human IgG1 heavy chain fragments are particularly prone to enhanced oxidation in periodontitis. PMID- 19280703 TI - Correction of 3D rigid body motion in fMRI time series by independent estimation of rotational and translational effects in k-space. AB - In functional magnetic resonance imaging (fMRI), even subvoxel motion dramatically corrupts the blood oxygenation level-dependent (BOLD) signal, invalidating the assumption that intensity variation in time is primarily due to neuronal activity. Thus, correction of the subject's head movements is a fundamental step to be performed prior to data analysis. Most motion correction techniques register a series of volumes assuming that rigid body motion, characterized by rotational and translational parameters, occurs. Unlike the most widely used applications for fMRI data processing, which correct motion in the image domain by numerically estimating rotational and translational components simultaneously, the algorithm presented here operates in a three-dimensional k space, to decouple and correct rotations and translations independently, offering new ways and more flexible procedures to estimate the parameters of interest. We developed an implementation of this method in MATLAB, and tested it on both simulated and experimental data. Its performance was quantified in terms of square differences and center of mass stability across time. Our data show that the algorithm proposed here successfully corrects for rigid-body motion, and its employment in future fMRI studies is feasible and promising. PMID- 19280704 TI - Genome-wide identification of cuticular protein genes in the silkworm, Bombyx mori. AB - Many kinds of cuticular proteins are found in a single insect species and their numbers and features are diversified among insects. Because there are so many cuticular proteins and so much sequence variation among them, an overview of cuticular protein gene is needed. Recently, a complete silkworm genome sequence was obtained through the integration of data from two whole genome sequence projects performed independently in 2004. To identify cuticular protein genes in the silkworm Bombyx mori exhaustively, we searched both the Bombyx whole genome sequence as well as various EST libraries, and found 220 putative cuticular protein genes. We also revised the annotation of the gene model, and named each identified cuticular protein based on its motif. The phylogenetic tree of cuticular protein genes among B. mori, Drosophila melanogaster, and Apis mellifera revealed that duplicate cuticular protein clusters have evolved independently among insects. Comparison of EST libraries and northern blot analyses showed that the tissue- and stage-specific expression of each gene was intricately regulated, even between adjacent genes in the same gene cluster. This study reveals many novel cuticular protein genes as well as insights into cuticular protein gene regulation. PMID- 19280705 TI - Determinants of platelet activation in hypertensives with microalbuminuria. AB - Microalbuminuria is a predictor of adverse outcome in hypertension.We evaluated in vivo platelet activation, by urinary 11-dehydrothromboxane (TX)B2 and plasma P selectin, in hypertensives with or without microalbuminuria, and its possible association with oxidative stress, by urinary 8-iso-prostaglandin (PG)F2alpha and endothelial dysfunction. Sixty essential hypertensive patients, with (n=30) or without (n=30) microalbuminuria, and 30 controls were studied. Endothelial function was assessed by nitric oxide products, intercellular adhesion molecule (ICAM)-1, and asymmetric dimethylarginine (ADMA) levels. Urinary 11-dehydro-TXB2 excretion was higher in microalbuminuric (median 805 pg/mg creatinine) compared to nonmicroalbuminuric patients or controls (414 and 291 pg/mg, respectively; P<0.0001). Plasma P-selectinwas significantly higher in patients with microalbuminuria (median 136 ng/ml) as compared to those without microalbuminuria or controls (85 and 65 ng/ml; P<0.0001). Urinary 8-iso-PGF2alpha excretion was also enhanced in microalbuminuric (median 279 pg/mg creatinine) compared to nonmicroalbuminuric patients or controls (157 and 146 pg/mg, respectively; P<0.0001). A significant impairment in endothelial function was found in microalbuminuric patients, with decreased nitric oxide and increased ICAM-1 and ADMA levels. Multivariate regression analysis showed that urinary 8-iso-PGF2alpha excretion (beta=0.49; P<0.0001) and microalbuminuria (beta=0.36; P<0.001) were independently related to 11-dehydro-TXB2 in hypertensives. Vitamin E supplementation (900 mg daily for 1 month) in 10 hypertensives with microalbuminuria was associated with normalization in median 11-dehydro-TXB2 and 8-iso-PGF2alpha. We conclude that lipid peroxidation is a major determinant of persistent platelet activation in hypertensive patients with microalbuminuria. PMID- 19280706 TI - BOLD correlates of EEG alpha phase-locking and the fMRI default mode network. AB - Phase locking or synchronization of brain areas is a key concept of information processing in the brain. Synchronous oscillations have been observed and investigated extensively in EEG during the past decades. EEG oscillations occur over a wide frequency range. In EEG, a prominent type of oscillations is alpha band activity, present typically when a subject is awake, but at rest with closed eyes. The spectral power of alpha rhythms has recently been investigated in simultaneous EEG/fMRI recordings, establishing a wide-range cortico-thalamic network. However, spectral power and synchronization are different measures and little is known about the correlations between BOLD effects and EEG synchronization. Interestingly, the fMRI BOLD signal also displays synchronous oscillations across different brain regions. These oscillations delineate so called resting state networks (RSNs) that resemble the correlation patterns of simultaneous EEG/fMRI recordings. However, the nature of these BOLD oscillations and their relations to EEG activity is still poorly understood. One hypothesis is that the subunits constituting a specific RSN may be coordinated by different EEG rhythms. In this study we report on evidence for this hypothesis. The BOLD correlates of global EEG synchronization (GFS) in the alpha frequency band are located in brain areas involved in specific RSNs, e.g. the 'default mode network'. Furthermore, our results confirm the hypothesis that specific RSNs are organized by long-range synchronization at least in the alpha frequency band. Finally, we could localize specific areas where the GFS BOLD correlates and the associated RSN overlap. Thus, we claim that not only the spectral dynamics of EEG are important, but also their spatio-temporal organization. PMID- 19280707 TI - The unique evolution of neuropeptide genes in the silkworm Bombyx mori. AB - Cloning-based approach combining homology search in the Bombyx genome sequence and Rapid Amplification of cDNA Ends (RACE) resulted in annotation of 23 neuropeptide genes and different splicing variants of three genes. In total 37 neuropeptide genes in addition to bombyxin gene family have been identified in Bombyx. Comparison of available insect neuropeptide genes revealed that the silkworm genome contains most conserved neuropeptide genes except those encoding proctolin, vasopressin-like peptide and neuropeptide-like precursor 2. In addition, we identified several paralogous neuropeptide genes which have not been found in other insects. The Bombyx genome contains a triplet of paralogous genes encoding adipokinetic hormones (AKH), two genes encoding different neuropeptide Fs (NPFs) and a tandem of related SIFamide and IMFamide genes. A novel gene coding for CCHamide was cloned and its expression in the CNS and midgut was demonstrated. Differential splicing was observed for the first time in transcripts for diuretic hormones and cardioacceleratory peptides 2b. Most paralogous genes or splicing variants of the same gene showed different expression patterns in the central nervous system (CNS). These results suggest that unique duplication and differential expression of several neuropeptide genes occurred during the evolution in Bombyx. This may be an effective mechanism for functional diversification of conserved neuropeptides. PMID- 19280708 TI - Common and distinct neural correlates of perceptual and memorial selection. AB - A critical aspect of cognitive control is the ability to select goal-relevant information in the face of competing distraction. A popular account is that common top-down selection processes underlie the ability to select among competing percepts and memories. We test the degree to which selective attention and memorial selection recruit the same neural resources. We demonstrate that both functions elicit largely overlapping networks within the dorsolateral prefrontal cortex (DLPFC), frontal eye fields (FEF), premotor cortex, and superior parietal lobule (SPL). Despite the close commonalities of selective attention and memorial selection, our results demonstrate that the SPL and FEF show preferential involvement in selective attention, whereas left ventrolateral prefrontal cortex (VLPFC) is uniquely associated with memorial selection. Thus, the two sorts of selection are not identical. We show further that variations in shared selection circuits are associated with differences in behavioral performance, suggesting that economy of control is beneficial to performance. PMID- 19280709 TI - Ethanol and tobacco smoke increase hepatic steatosis and hypoxia in the hypercholesterolemic apoE(-/-) mouse: implications for a "multihit" hypothesis of fatty liver disease. AB - Although epidemiologic studies indicate that combined exposure to cigarette smoke and alcohol increase the risk and severity of liver diseases, the molecular mechanisms responsible for hepatotoxicity are unknown. Similarly, emerging evidence indicates a linkage among hepatic steatosis and cardiovascular disease. Herein, we hypothesize that combined exposure to alcohol and environmental tobacco smoke (ETS) on a hypercholesterolemic background increases liver injury through oxidative/nitrative stress, hypoxia, and mitochondrial damage. To test this, male apoE(-/-) mice were exposed to an ethanol-containing diet, ETS alone, or a combination of the two, and histology and functional endpoints were compared to filtered-air-exposed, ethanol-naive controls.Whereas ethanol consumption induced a mild steatosis, combined exposure to ethanol + ETS resulted in increased hepatic steatosis, inflammation, alpha-smooth muscle actin, and collagen. Exposure to ethanol + ETS induced the largest increase in CYP2E1 and iNOS protein, as well as increased 3-nitrotyrosine, mtDNA damage, and decreased cytochrome c oxidase protein, compared to all other groups. Similarly, the largest increase in HIF1alpha expression was observed in the ethanol + ETS group, indicating enhanced hypoxia. These studies demonstrate that ETS increases alcohol dependent steatosis and hypoxic stress. Therefore, ETS may be a key environmental "hit" that accelerates and exacerbates alcoholic liver disease in hypercholesterolemic apoE(-/-) mice. PMID- 19280710 TI - Identification, genomic organization and expression pattern of glutathione S transferase in the silkworm, Bombyx mori. AB - Glutathione S-transferases (GSTs) are a multifunctional supergene family and some play an important role in insecticide resistance. We have identified 23 putative cytosolic GSTs by searching the new assembly of the Bombyx mori genome sequence. Phylogenetic analyses on the amino acid sequences reveal that 21 of the B. mori GSTs fall into six classes represented in other insects, the other two being unclassified. The majority of the silkworm GSTs belong to the Delta, Epsilon, and Omega classes. Most members of each class are tandemly arranged in the genome, except for the Epsilon GSTs. Expressed sequence tags (ESTs) corresponding to 19 of the 23 GSTs were found in available databases. Furthermore RT-PCR experiments detected expression of all the GSTs in multiple tissues on day 3 of fifth instar larvae. Surprisingly, we found little or no expression of most Delta and Epsilon GSTs in the fat body, which is thought to be the main detoxification organ. This may explain the sensitivity of the silkworm to certain insecticides. Our data provide some insights into the evolution of the B. mori GST family and the functions of individual GST enzymes. PMID- 19280711 TI - Anterior insular cortex anticipates impending stimulus significance. AB - Touch is a fundamental, but complex, element of everyday interaction that impacts one's sensory and affective experience via interoceptive processing. The insular cortex is an integral component of the neural processes involved in interoception, i.e. the generation of an "emotional moment in time" through the sensing of the internal body state (Craig, A.D., 2002. How do you feel? Interoception: the sense of the physiological condition of the body. Nat. Rev. Neurosci 3, 655-666.). Here, we examine the contribution of different parts of the insular cortex in the representation of both affective and sensory aspects of touch. To that end, subjects were administered a cued application of touch during functional MRI. We find that stimulus-related activation occurs in the mid-to posterior insula, whereas anticipatory related activation is seen mostly in anterior insula. Moreover, the degree of activation in anterior insula during anticipation is correlated with the degree of activation in the posterior insula and caudate during stimulus processing. Finally, the degree of activation in the anterior insula during anticipation is also correlated with experienced intensity of the touch. Taken together, these results are consistent with the hypothesis that the anterior insula is preparing for the sensory and affective impact of touch. This preparatory function has important implications for the understanding of both anxiety and addictive disorders because dysfunctions in anticipatory processing are a fundamental part of the psychopathology. PMID- 19280712 TI - The tolerogenic peptide hCDR1 downregulates pathogenic cytokines and apoptosis and upregulates immunosuppressive molecules and regulatory T cells in peripheral blood mononuclear cells of lupus patients. AB - A tolerogenic peptide, hCDR1, ameliorated murine lupus via the upregulation of functional regulatory cells and by immunomodulating cytokine production. In the present study we analyzed the ability of hCDR1 to similarly affect gene expression and regulatory T cells when incubated with peripheral blood mononuclear cells (PBMC) of lupus patients. To this end, peripheral blood mononuclear cells (PBMC) of 11 lupus patients and five gender- and age-matched healthy controls were cultured with hCDR1 or a control peptide. Gene expression and regulatory T-cells were assessed. hCDR1 significantly downregulated interleukin (IL)-1beta, interferon (IFN)-gamma, and IL-10 gene expression. Furthermore, hCDR1 upregulated the expression of the anti-apoptotic Bcl-xL molecule and downregulated the pro-apoptotic caspase-3, resulting in reduced rates of apoptosis. hCDR1 increased the expression of transforming growth factor (TGF)-beta, FoxP3 and the negative regulators Foxj1 and Foxo3a. No significant effects were observed using a control peptide or when PBMC of healthy donors were incubated with hCDR1. The elevated gene expression of FoxP3 was due to hCDR1 induced upregulation of TGF-beta, resulting in an increase of CD4+CD25+FoxP3+ functional, regulatory cells. The ability of the regulatory cells to diminish IFN gamma expression and to upregulate TGF-beta was abrogated after the addition of a neutralizing anti-CD25 antibody, confirming their role in the beneficial effects of hCDR1. PMID- 19280713 TI - Reactive oxygen species regulation by AIF- and complex I-depleted brain mitochondria. AB - Apoptosis-inducing factor (AIF)-deficient harlequin (Hq) mice undergo neurodegeneration associated with a 40-50% reduction in complex I level and activity. We tested the hypothesis that AIF and complex I regulate reactive oxygen species (ROS) production by brain mitochondria. Isolated Hq brain mitochondria oxidizing complex I substrates displayed no difference compared to wild type (WT) in basal ROS production, H2O2 removal, or ROS production stimulated by complex I inhibitors rotenone or 1-methyl-4-phenylpyridinium. In contrast, ROS production caused by reverse electron transfer to complex I was attenuated by approximately 50% in Hq mitochondria oxidizing the complex II substrate succinate. Basal and rotenone-stimulated rates of H2O2 release from in situ mitochondria did not differ between Hq and WT synaptosomes metabolizing glucose, nor did the level of in vivo oxidative protein carbonyl modifications detected in synaptosomes, brain mitochondria, or homogenates. Our results suggest that AIF does not directly modulate ROS release from brain mitochondria. In addition, they demonstrate that in contrast to ROS produced by mitochondria oxidizing succinate, ROS release from in situ synaptosomal mitochondria or from isolated brain mitochondria oxidizing complex I substrates is not proportional to the amount of complex I. These findings raise the important possibility that complex I contributes less to physiological ROS production by brain mitochondria than previously suggested. PMID- 19280714 TI - Cigarette smoke extract induces cytosolic phospholipase A2 expression via NADPH oxidase, MAPKs, AP-1, and NF-kappaB in human tracheal smooth muscle cells. AB - Up-regulation of cytosolic phospholipase A2 (cPLA2) by cigarette smoke extract (CSE) may play a critical role in airway inflammatory diseases. However, the mechanisms underlying CSE-induced cPLA2 expression in human tracheal smooth muscle cells (HTSMCs) remain unknown. CSE induced cPLA2 protein and mRNA expression, and ROS generation was attenuated by pretreatment with a reactive oxygen species (ROS) scavenger (N-acetylcysteine), or inhibitors of NADPH oxidase (diphenyleneiodonium chloride, apocynin) and transfection with p47phox siRNA, suggesting that CSE-induced cPLA2 expression was mediated through NADPH oxidase activation and ROS production in HTSMCs. Furthermore, CSE-induced cPLA2 expression was attenuated by pretreatment with the inhibitors of MEK1/2 (U0126), p38 MAPK (SB202190), and JNK (SP600125), which were further confirmed by transfection with siRNAs of JNK1, p42, and p38 to down-regulate the expression of respective proteins and reduce cPLA2 expression. Induction of cPLA2 by CSE was attenuated by selective inhibitors of NF-kappaB (helenalin) and AP-1 (curcumin). Moreover, promoter assays revealed that increases of cPLA2, NF-kappaB, and AP-1 luciferase activities stimulated by CSE were attenuated by these inhibitors. These results suggest that in HTSMCs, CSE induced NADPH oxidase activation leading to phosphorylation of p42/p44 MAPK, p38 MAPK, and JNK. These reactions induced nuclear transcription NF-kappaB and AP-1 activities which were essential for CSE-induced cPLA2 gene expression. PMID- 19280715 TI - Predisposition to idiopathic thrombocytopenic purpura maps close to the major histocompatibility complex class I chain-related gene A. AB - Idiopathic thrombocytopenic purpura (ITP) is an autoimmune condition with poorly known etiology, characterized by platelet destruction. Genetic association studies of this disease are scarce, discrepant, and restricted to major histocompatibility complex (MHC) polymorphisms. Hence, a case-control study was conducted with an aim to map the MHC to IPT susceptibility using HLA-B and nine microsatellite loci encompassing MHC class I, II, and III regions. We compared the allelic frequencies in samples of unrelated healthy controls and ITP patients. After correction for multiple tests, only allele MICA*183, also known as A5.1, demonstrated an association, resulting in the identification of a major predisposing region close to STR-MICA. This result may highlight the putative functional role of MICA in the immune response to ITP. PMID- 19280716 TI - The IL6-174G/C polymorphism is associated with celiac disease susceptibility in girls. AB - The aim of this paper was to study the role of IL6 and IL6R polymorphisms in celiac disease (CD) susceptibility. Because previous literature describes IL-6 related gender differences, sex-stratified analyses were performed. We undertook a case-control study with 374 pediatric CD patients and 853 healthy controls, all white Spaniards, and a family study using an independent sample including 303 trios for replication purposes. Three single-nucleotide polymorphisms tagging most of the variability of the IL6 gene (rs2069827, rs1800795 [-174G/C], and rs2069840) and one functional polymorphism in IL6R (rs8192284, Asp358Ala) were genotyped using TaqMan technology. Case-control comparisons were performed with the chi2 test and family data were analyzed with the transmission disequilibrium test. No association was observed between any tested polymorphism and overall CD. However, after sex stratification, we found that the IL6 promoter variant -174C increases the risk of developing CD specifically in female patients. This effect was observed both in the case-control and in the family studies (considering girls included in both studies vs boys: p = 0.021, odds ratio [OR] = 1.31, 95% confidence interval [CI] 1.03-1.66; and vs controls: p = 0.003, OR = 1.30, 95% CI 1.09-1.55). The functional -174G/C IL6 polymorphism seems to influence CD susceptibility in girls. The gender-specific role of IL-6 in this pathology must be further investigated. PMID- 19280717 TI - Microsatellite polymorphism in intron 2 of human Toll-like receptor 2 gene is associated with susceptibility to acute pancreatitis in Japan. AB - This study evaluated the association of the polymorphisms in the Toll-like receptor (TLR)2 and TLR4 genes with acute pancreatitis (AP) in Japan. The numbers of guanine-thymine [(GT)n] repeats in intron 2 of the TLR2 gene were counted in 202 unrelated patients with AP (80 with severe and 122 with mild disease) and in 286 healthy controls, using polymerase chain reaction and Genescan analysis. The alleles were divided into three subclasses: (GT)16 or less as the S allele; between (GT)17 and (GT)22 as the M allele; and (GT)23 or more as the L allele. Asp299Gly and Thr399Ile polymorphisms in the TLR4 gene were examined by polymerase chain reaction-restriction fragment length polymorphism analysis. Patients with AP had more S alleles (p < 0.001; odds ratio = 2.37; 95% confidence interval = 1.78-3.17) and fewer M alleles (p < 0.001; odds ratio = 0.40; 95% confidence interval 0.31-0.52) than did healthy controls. Genotypes SS and SL were more common, whereas MM and ML were less common in patients with AP. In subgroup analyses, the genotypes including S alleles were more common in patients with severe AP than in controls. No Asp299Gly and Thr399Ile polymorphisms were detected. In conclusion, microsatellite polymorphism in intron 2 of the TLR2 gene was associated with susceptibility to AP and its severity in Japan. PMID- 19280718 TI - Leukotriene C4 synthase and ischemic cardiovascular disease and obstructive pulmonary disease in 13,000 individuals. AB - Ischemic cardiovascular disease and obstructive pulmonary disease involve inflammation. Leukotrienes may be important pro-inflammatory mediators. We tested the hypothesis that the (-1072)G > A and (-444)A > C promoter polymorphisms of leukotriene C4 synthase confer risk of transient ischemic attack (TIA), ischemic stroke, ischemic heart disease (IHD), asthma, and chronic obstructive pulmonary disease (COPD). We genotyped individuals from the Danish general population, the Copenhagen City Heart Study, and Danish patients with IHD/coronary atherosclerosis, the Copenhagen Ischemic Heart Disease Study. We used prospective (n = 10,386), cross-sectional (n = 10,386), and case-control (n = 2392 + 5012) designs. Allele frequency was 0.07 for (-1072)A and 0.29 for (-444)C. Cumulative incidence for TIA was higher for (-1072)AA versus GG genotype (log-rank: p < 0.001), and lower for (-444)CC versus AA genotype (log-rank: p = 0.03). Cumulative incidence for ischemic stroke was also lower for (-444)CC versus AA genotype (log-rank: p = 0.04). Multifactorially adjusted hazard ratios for TIA were 5.2(95% CI:1.9-14) for (-1072)AA versus GG genotype, and 0.4(0.2-1.0) for ( 444)CC versus AA genotype. Corresponding values were 1.9 (0.7-5.2) and 0.7 (0.5 1.0) for ischemic stroke, and 0.8 (0.4-1.6) and 1.0 (0.9-1.2) for IHD. In the case-control study, corresponding multifactorially adjusted odds ratios for IHD/coronary atherosclerosis were 0.5 (0.2-1.3) and 1.2 (1.0-1.5). These genotypes were not associated with risk of asthma or COPD. Leukotriene C4 synthase promoter genotypes influence risk of TIA and ischemic stroke, but not risk of IHD/coronary atherosclerosis, asthma, or COPD. PMID- 19280719 TI - Intestinal absorption of long-chain fatty acids: evidence and uncertainties. AB - Over the two last decades, cloning of proteins responsible for trafficking and metabolic fate of long-chain fatty acids (LCFA) in gut has provided new insights on cellular and molecular mechanisms involved in fat absorption. To this systematic cloning period, functional genomics has succeeded in providing a new set of surprises. Disruption of several genes, thought to play a crucial role in LCFA absorption, did not lead to clear phenotypes. This observation raises the question of the real physiological role of lipid-binding proteins and lipid metabolizing enzymes expressed in enterocytes. The goal of this review is to analyze present knowledge concerning the main steps of intestinal fat absorption from LCFA uptake to lipoprotein release and to assess their impact on health. PMID- 19280720 TI - A sentinel event for Ontario hospitals. PMID- 19280721 TI - Advocate or abdicate: the responsible choice for today's emergency medicine resident. PMID- 19280722 TI - Does fluoxetine restore plasticity in the rat retina? PMID- 19280723 TI - Simply operational. Surgical productivity at Lahey Clinic gets a boost from business activity monitoring and role-based dashboards. PMID- 19280724 TI - [Results and prospects of scientific research on sensory conflict under exposure to noise and vibration at work]. AB - The article presents results of longstanding research concerning influence of physical occupational hazards on workers' health. The authors justify a concept of sensory conflict developing under exposure to physical hazards and being a key chain in occupational diseases formation. New approaches are suggested to prevent occupational diseases caused by physical factors. PMID- 19280725 TI - [Topographic mapping in comparative evaluation of brain electric activity in individuals with occupational vs. somatic diseases]. AB - The authors presented results of topographic mapping and comparative evaluation of EEC patterns in patients with occupational and non-occupational diseases, apparently healthy individuals. Findings are unidirectional changes in EEC parameters among occupational diseases patients. PMID- 19280726 TI - [Relations of level of engineer's health with professional creative activity]. AB - Risks of the main pathologic syndromes, social-psychological characteristics, value-motivational sphere and manifestation of burnout in engineers-designers of flying devices and electronics with a different degree of expressiveness of professional creative activity are investigated. It is shown, that external social-psychological are factors as determinants of burnout and risks of health deterioration, prevalence of symptoms of burnout depend on a degree of creative activity. PMID- 19280727 TI - [Morphologic and neurochemical effects in long-term period of mercurial intoxication (experimental data)]. AB - The article deals with influence of longstanding inhalation of mercurial vapors on behavior of white rats, on morphologic and functional disorders in nervous system, on peculiarities of neuromediators metabolism in brain homogenate over the post-contact period. PMID- 19280728 TI - Nourishing inmates in Malagasy prisons. PMID- 19280729 TI - Standards of neuroscience nursing practice. PMID- 19280730 TI - Dengue in Africa: emergence of DENV-3, Cote d'Ivoire, 2008. PMID- 19280731 TI - Meeting of the International Task Force for Disease Eradication--29 October 2008. PMID- 19280732 TI - Antimalarial drug resistance, Thai-Cambodian border. PMID- 19280733 TI - Why "race" is a central idea in Australia's construction of the idea of a nation. PMID- 19280734 TI - Case of the month. Squamous cell carcinoma, keratoacanthoma type. PMID- 19280735 TI - Reaching beyond the rainbow to provide healthy smiles. PMID- 19280736 TI - Bringing dental literacy to the classroom. PMID- 19280737 TI - The "Smitherman touch". PMID- 19280738 TI - "With a little bit of luck": coping with adjustment in urban Ghana, 1975-90. PMID- 19280739 TI - Women and culture in Poland in early modern times. PMID- 19280740 TI - We mourn our great loss. Paying homage to Martin Delaney. 1945-2009. PMID- 19280741 TI - HIV drugs in the pipeline. A concise summary of drugs further along in development. PMID- 19280742 TI - [From the history of medical students' dances]. PMID- 19280743 TI - HIV drug guide introduction. A brief description of the drug classes and how they work. PMID- 19280744 TI - Patient assistance and drug co-pay programs. PMID- 19280745 TI - 13th annual HIV drug guide. PMID- 19280746 TI - Drug interactions chart. An abbreviated, at-a-glance guide to HIV drug interactions. PMID- 19280747 TI - Side effects chart. An abbreviated, at-a-glance guide to potential HIV drug side effects. PMID- 19280748 TI - Current DHHS treatment guidelines for first time therapy. PMID- 19280749 TI - Borgu in the Atlantic slave trade. PMID- 19280750 TI - Chinese-Thai transmigrants: reworking identities and gender relations in Thailand and the United States. PMID- 19280751 TI - Collectivization in the 1970s and 1980s in Zamagurie, Slovakia. PMID- 19280752 TI - Culture, kinship, and power: the evolution of royal slavery in nineteenth-century Kano. PMID- 19280753 TI - Exorcising witchcraft: the return of the gods in new religious movements on Jos Plateau and the Benue regions of Nigeria. PMID- 19280754 TI - Evolutionary genetics: 150 years of natural selection. Preface. PMID- 19280755 TI - Lung fibrosis and lung cancer incidence in beagle dogs that inhaled 238PuO2 or 239PuO2. AB - Determination of radiation protection guidelines for persons working with plutonium has been complicated by limited human data on the biological behavior and subsequent health effects from internally deposited plutonium. One solution has been the use of animal models to predict likely health effects in humans. To compare the relationships between plutonium inhalation and lung fibrosis and lung cancer, data from life-span studies of beagle dogs given a single exposure to either plutonium-238 dioxide (238PuO2) or plutonium-239 dioxide (239PuO2) were analyzed. Estimates of the cumulative hazard of lung fibrosis and lung cancer after exposure to either were generated. The hazard of lung fibrosis was not consistent with a linear no-threshold model, although the magnitude of the threshold differed by radionuclide. In dogs given 239PuO2, the best model of lung fibrosis incorporated a linear dose-response function; a linear-quadratic dose response function fit the data better in dogs given 238PuO2. At any given cumulative dose, the lung fibrosis hazard was greater for dogs given 238PuO2. In dogs given 238PuO2, with or without covariates, a quadratic dose-response function for lung cancer hazard fit better than a linear no-threshold model. In dogs given 239PuO2, models of lung cancer with the dose-response function as the sole predictor variable were consistent with a linear no-threshold model; however, a quadratic dose-response function with a cell-killing term fit better. These findings have implications for radiation protection because, while lung cancer hazard was dependent on cumulative dose, regardless of isotope, the lung fibrosis hazard depended on both cumulative dose and isotope. PMID- 19280757 TI - From petty traders to international merchants: a historical account of three Igbo women of Nigeria in trade and commerce, 1886 to 1870. PMID- 19280758 TI - Gender, medicine, and consumer culture in Victorian England: creating the kleptomaniac. PMID- 19280759 TI - Green dots, pink hearts: displacing politics from the Malaysian rain forest. PMID- 19280760 TI - America's "other" drug problem. PMID- 19280761 TI - [The salon of Koidula: a living political friendship circle]. PMID- 19280762 TI - Screening for obstructive sleep apnea during commercial driver medical examinations. AB - OBJECTIVE: To evaluate consensus criteria for screening commercial drivers for obstructive sleep apnea (OSA). METHODS: Consecutive drivers underwent OSA screening using Joint Task Force consensus criteria at commercial driver medical examinations. Outcomes included: clinical yield of screening; and drivers' compliance with polysomnography (PSG) referrals and OSA treatment. RESULTS: Among 456 drivers, 53 (12%) were referred for PSG, and 20/20 were confirmed to have OSA, supporting a high positive predictive value. The other 33 drivers referred for PSG were lost to follow-up but demonstrated no significant differences from those with confirmed OSA. After diagnosis, only one of 20 drivers with confirmed OSA demonstrated treatment compliance. CONCLUSIONS: Drivers identified by the consensus criteria have a high likelihood of OSA. Drivers' poor compliance with PSGs and OSA treatment support federally mandated screening of commercial drivers. PMID- 19280763 TI - Seasonal influenza prevention in health care workers. PMID- 19280764 TI - Hypoalphalipoproteinemia in populations of Native American ancestry: an opportunity to assess the interaction of genes and the environment. AB - PURPOSE OF THIS REVIEW: Our aim is to review the environmental and genetic factors associated with hypoalphalipoproteinemia in populations of Native American ancestry. We examine the strength of the association and outline the population-specific genetic factors that lead to a higher susceptibilty for this condition. RECENT FINDINGS: Low HDL is the most common lipid abnormality in populations of Native American ancestry. Population-based surveys carried out in Latin America and in Mexican Americans shows that 40-60% of adults have hypoalphalipoproteinemia. The contribution of this trait to the metabolic syndrome is greater in individuals with Native American ancestry than in other ethnic groups. Several environmental factors have contributed to this phenomenon (i.e. high dietary content of carbohydrates and fat due to cultural factors and a growing incidence of obesity). In addition, results from recent genetic studies show that certain hypoalphalipoproteinemia susceptibility alleles are ethnic specific for Native Americans. The variant R230C of the ATP-binding cassette transporter subfamily A member 1 gene (ABC-A1) is common among mestizos (10.9% in Mexican mestizos) and its presence has a significant negative effect on HDL cholesterol levels (-4.2%). An additional noteworthy finding is that the R230C variant appears to be specific for the Amerindian populations. Its allele frequency is 0.28 in Mayans, 0.214 in Purepechas, 0.203 in Yaquis and 0.179 among Teenek. In contrast, the C230 allele has not been found in African, European, Chinese or South Asian populations. SUMMARY: The assessment of the genetic and environmental determinants of hypoalphalipoproteinemia in populations of Native American origin provides an opportunity to assess the population-specific interactions between genes and the environment PMID- 19280765 TI - New estimates of land productivity in Belgium, 1750-1850. PMID- 19280766 TI - Hepatic nuclear factor 1-alpha: inflammation, genetics, and atherosclerosis. AB - PURPOSE OF REVIEW: Increased plasma levels of C-reactive protein (CRP), a hepatic acute phase reactant, predict risk for coronary heart disease. There has been interest in identifying genetic determinants of CRP as a means of better understanding its regulation and its relation to coronary heart disease. We here review recent findings that have linked plasma CRP levels to single nucleotide polymorphisms in hepatic nuclear factor (HNF) 1-alpha, a transcription factor with a wide range of functions, including many involved in cholesterol, bile acid, and lipoprotein metabolism. RECENT FINDINGS: Two genome-wide association studies have identified single nucleotide polymorphisms in several genes that are strongly related to plasma CRP levels, including several on chromosome 12 in the vicinity of the HNF1A gene. The CRP gene promoter has two HNF1-alpha-binding sites. Recently, it has been demonstrated that HNF1-alpha is required for cytokine-driven CRP expression and that this involves formation of a complex with STAT3 and c-Fos. SUMMARY: Based on the recent genetic findings as well as delineation of the role of HNF1-alpha in regulating the expression of the CRP gene, it appears that this transcription factor may play a key role in linking metabolic and inflammatory pathways underlying the pathogenesis of coronary heart disease. PMID- 19280767 TI - "Real Belizean food": building local identity in the transnational Caribbean. PMID- 19280768 TI - Sensational stories, endangered bodies: women's work and the new journalism in England in the 1890s. PMID- 19280769 TI - Atypical chest pain in athletes. AB - Initial management of chest pain in athletes always should involve assessment for serious, life-threatening causes, such as myocardial infarction. However, atypical chest pain, or chest pain not due to myocardial ischaemia, is a common presentation in the athletic population. This review looks at the possible causes of atypical chest pain in athletes, focusing upon conditions that are more common in athletes than the general population or that have a link to exercise. Causes can be grouped due to the system involved (musculoskeletal, gastrointestinal, respiratory, cardiac) with more common causes including rib stress fractures, costochondritis, muscle strain, gastroesophageal reflux, and exercise-induced asthma. Psychogenic causes can be common in children/adolescents. Return to play is discussed, with some conditions such as myocarditis warranting a long (at least 6 months) absence from training, whereas others such as precordial catch require nothing more than reassurance. PMID- 19280770 TI - [Gastrointestinal oncology. Is endoscopic follow-up required for the early detection of primary gastric cancer in patients with gastrectomy due to benign disease?]. PMID- 19280771 TI - [Functional gastrointestinal disorders. Patient with pyrosis showing no improvement with a high-dose proton pump inhibitor. What is the diagnosis and treatment?]. PMID- 19280772 TI - [Pancreas and biliary tree. Should parenteral, nasogastric enteral or nasojejunal enteral nutrition be used in severe acute pancreatitis?]. PMID- 19280773 TI - [Inflammatory bowel disease. If a patient with Crohn's disease remains in remission for 4 years with azathioprine (or mercaptopurine), should the drug be withdrawn?]. PMID- 19280774 TI - [Gastrointestinal endoscopy. In patients with jaundice secondary to unresectable carcinoma of the pancreatic head, what is the endoscopic treatment of choice to alleviate the jaundice?]. PMID- 19280775 TI - [Acid-related diseases. How useful is impedancometry in the diagnosis of gastroesophageal reflux disease?]. PMID- 19280776 TI - [Malnutrition in children with DAMP?]. PMID- 19280779 TI - Laser dentistry: an introduction. PMID- 19280780 TI - Lasers in pediatric practice. PMID- 19280781 TI - Laser dentistry. Modern technology or newest fad? PMID- 19280782 TI - Mission dentistry. Know the drill! PMID- 19280783 TI - The time of determination of gynoparae and males in the bird cherry-oat aphid, Rhopalosiphum padi. AB - Apterous exules of Rhopalosiphum padi which experienced short photoperiods only during the first half of their nymphal life all produced gynoparae which made up 57% of the offspring produced in the first 7 days of the mother's reproductive life. Short photoperiods during the second half of a mother's nymphal life did not induce the production of gynoparae. However, when short photoperiods were experienced throughout a mother's nymphal life significantly more gynoparae (82%) were produced. Ten per cent of the offspring of mothers that experienced short photoperiods only during their adult life developed into gynoparae. Of the offspring reared in short photoperiods, but born to mothers reared and kept in long photoperiods, 30% developed into gynoparae. When mothers were exposed to short photoperiods in their adult life and their offspring so treated, then 90% of the offspring developed into gynoparae. Males only occurred amongst the last offspring of mothers that experienced short photoperiods in either the first or the second half of their nymphal life. PMID- 19280784 TI - Susceptibility of plants to attack by wireworms (Agriotes spp.). AB - In laboratory tests, larvae of Agriotes spp. were presented with a choice between seedlings of test plants, and of wheat, which is known to be susceptible. Some plants, e.g. onion, were as susceptible as wheat to wireworm attack whilst others (mustard, cabbage, French marigold, clover and flax) were less often attacked, partly because they are small and are shallow sown. All pea and bean plants exposed to wireworms were attacked but most tolerated attack and continued to grow. PMID- 19280785 TI - Intraspecific variation of Myzus persicae on sugar beet (Beta vulgaris). AB - Differences in inherited resistance among seven sugar-beet stocks had similar effects on Myzus persicae clones representing the range of variation in aphid response to resistant and susceptible sugar beet observed in fifty-eight clones collected between 1969 and 1971. Three sugar-beet stocks were consistently resistant. Statistically significant interactions between beet stocks and aphid clones did not indicate the existence of biotypes with specific abilities to overcome resistance. M. persicae clones differed in their vigour of colonizing sugar beet, irrespective of the differences between beet stocks. The readiness of adult aphids to settle determined the size of aphid population produced and included a component related to the response of the aphid clone to sugar beet as a host, and a component related to the resistance ranking of the beet stock. Breeding sugar beet with resistance to aphids will be simplified, as the results indicate that, at present, differences between aphid biotypes need not be considered a problem. PMID- 19280786 TI - Supercooling and the low-temperature survival of the green spruce aphid Elatobium abietinum. AB - Laboratory investigations into the low-temperature tolerance of the green spruce aphid, Elatobium abietinum, revealed that the insect was killed by freezing. Aphids and host Sitka spruce needles showed similar seasonal changes in supercooling ability. A noticeable increase in this ability occurred between June and October. Aphids were more susceptible to low temperatures when attached to the plant. It is suggested that mortality resulted from ice which formed in the sap of the host needles and spread into the feeding aphids via their mouthparts. Neither the chlorotic banding of needles, caused by aphid feeding, nor needle length affected needle supercooling. Increased duration of exposure increased the probability of freezing of supercooled needles at low temperatures. A small percentage of first-instar nymphs supercooled to much lower temperatures than the remainder of the population. These were newly born nymphs whose high supercooling ability markedly decreased when they began to feed. PMID- 19280787 TI - The effect of store conditions on the rotting of apples, cv. Bramley's Seedling, by Nectria galligena. AB - The log of the time interval between inoculation with Nectria galligena in October and the onset of rotting in apples held in air was proportional to the deficit between the temperature of incubation and 25 degrees C, but temperature did not affect the rate of subsequent rot expansion. Rots expanded equally fast whether apples were held in dry or moist air. The quantity of rotted tissue obtained after incubating inoculated apples in atmospheres containing up to 12.5% CO2 increased with increasing concentrations of CO2 greater than 2.5%. The quantity of rotted tissue obtained in apples incubated in 10% CO2 was three times as great as that obtained after incubation in air. The incidence of natural rots was lower in apples stored at 4% CO2 than in those stored in air and rotting increased with increasing concentrations of CO2 higher than 4%. Colonies of N. galligena grew faster on malt agar plates incubated in 5% CO2 than in air, but growth was slower in 10% CO2 than in air. The quantity of benzoic acid per mg hyphae accumulated in developing lesions was similarly related to the CO2 concentrations up to 2.5% but decreased at higher concentrations, and the quantities found in apples stored in CO2 concentrations > 5.0% CO2 were less than in those stored in air. PMID- 19280788 TI - Spiroplasmas are the causal agents of citrus little-leaf disease. AB - A spiroplasma isolated from citrus with little-leaf disease was grown in a cell free medium and injected into leafhoppers (Euscelis plebejus). Injected leafhoppers, but not those fed on infected plants, transmitted the spiroplasma to white clover (Trifolium repens cv. S100) and sweet orange (Citrus sinensis cv. Valencia). Infected clover plants were severely stunted; infected sweet orange plants showed typical symptoms of citrus little-leaf disease. The spiroplasma was detected in clover and sweet orange plants by electron microscopy; the helical morphology of the organisms was most easily recognizable in sections 150-200 nm thick. The organism was re-isolated in cell-free media both from infected plants and from injected E. plebejus. The original isolate and those re-isolated from experimentally infected clover and sweet orange appeared by morphological, cultural, biochemical and serological criteria to be identical to each other and to the R8-A2 (type) and C-189 strains of Spiroplasma citri. Serological tests and electrophoretic analysis of protein preparations indicated no relationship to Acholeplasma laidlawii, although this organism survived for at least 10 wk after injection into E. plebejus. Our results show that the causal agent of little-leaf disease is related to S. citri. PMID- 19280789 TI - Investigations on fungicides. XVI. Natural resistance of plant roots to fungal pathogens. AB - Uninoculated roots of pea (Pisum sativum) and French bean (Phaseolus vulgaris) have been shown to exude a number of antifungal compounds when grown in a non sterile aqueous aerated medium. These have been identified and their possible importance in relation to disease resistance is discussed. PMID- 19280790 TI - Sugar, organic acid, phenolic acid and plant growth regulator content of extracts of honeydew of the aphid Myzus persicae and of its host plant, Raphanus sativus. AB - The honeydew of Myzus persicae feeding on radish seedlings contains glucose, fructose, trehalose, melezitose and sucrose, of which glucose and fructose are present in radish seedlings. In addition, the honeydew contains eight organic acids, of which seven are present in uninfested seedlings and six are present in infested radish seedlings: of the eighteen phenolic acids in the honeydew, five are present in uninfested radish seedlings. Auxins, gibberellins, growth inhibitors and cytokinins are present in the honeydew. The presence of carbonate and bicarbonate previously recorded in honeydew of M. persicae could not be confirmed, but ammonia is present in fresh honeydew and accounts for its high pH. PMID- 19280791 TI - The influence of expanding leaves and the reproductive stem apex on apical dominance in Lolium multiflorum. AB - In vegetative plants of Lolium multiflorum removal of the two youngest emerging leaves resulted in increased expansion of basal tiller buds. A similar release of inhibition of tiller buds took place if the floriferous apex was removed. The surgical procedures did not affect the response. Under conditions of N-deficiency total tiller number was reduced but on removal of the apex the deficient plants showed an increased initial rate of tiller bud expansion. Apical dominance during the vegetative stage of growth in this grass was apparently due to the expanding leaves in the vegetative apex, but in the flowering plant the control was exerted by the inflorescence or the elongating stem. PMID- 19280792 TI - A model for the mechanism of stimulation of latex flow in Hevea brasiliensis by ethylene. AB - A model is proposed for ethylene stimulation of latex flow from tapping cuts of Hevea brasiliensis, based on the predicted effects of ethylene on the plasticity and structure of cell walls of the latex vessels. Evidence for the model comes from the effects of ethylene on etiolated shoots of Pisum sativum. PMID- 19280793 TI - Resistance to bacterial blight of cotton varieties homozygous for combinations of B resistance genes. AB - In the Sudan, inoculation experiments were done on a wide range of Egyptian cotton lines homozygous for single genes and for digenic and trigenic combinations of Knight's B genes for resistance to bacterial blight. Although leaf inoculation was successful, stem inoculation was only partially so and boll inoculation, using two different techniques, failed to produce measurable disease symptoms. There was a good general relationship between leaf and stem resistance, and a close association between resistance to natural attack in the field and leaf resistance to artificial inoculation. Confirmation was obtained of the strong resistance conferred by B2B9K, which was as effective as B2B6 or B2B3B6. No other combination was as effective when inoculated artificially. Nevertheless, in a natural field attack, only mild symptoms were found in lines homozygous for B1B9K, and for B4B6. Under the same conditions lines with B2B6 showed no symptoms but those with B2B3 were severely attacked. Reasons for continuing to use major genes in the breeding of blight-resistant cotton in the Sudan are discussed. PMID- 19280794 TI - The diagnostic value of ultrasonography in patients with electrophysiologicaly confirmed carpal tunnel syndrome. AB - OBJECTIVE: To evaluate the diagnostic value of ultrasonography in patients with electrophysiologically confirmed carpal tunnel syndrome. DESIGN: A prospective ultrasonographic study of 70 wrists with electrophysiologically confirmed carpal tunnel syndrome and of 80 normal wrists. Receiver-operating-characteristics curves for the ultrasonographic measurements of median nerve were plotted to identify the most optimal cutoff values. RESULTS: The ultrasonographic measurements of median nerves were found to be increased significantly in patients with carpal tunnel syndrome when compared with controls, particularly in terms of cross-sectional area (P <0.001). According to receiver-operating characteristics curve results, the most optimal cutoff value for the cross sectional area of the median nerve was obtained at the level of middle carpal tunnel, which was 9.3 mm2, with a sensitivity of 80% and specificity of 77.5%. CONCLUSION: Ultrasonographic examination of the median nerve seems to be a promising method in the diagnosis of carpal tunnel syndrome, evaluating the morphologic changes of the median nerve in patients with clinical signs and symptoms. Further studies with wider series are needed to confirm our preliminary results. PMID- 19280795 TI - Patellofemoral pain and the mechanomyographic responses of the vastus lateralis and vastus medialis muscles. AB - The purpose of this investigation was to examine the influence of patellofemoral pain (PFP) on the amplitude of the mechanomyographic (MMG) and electromyographic (EMG) signals from the vastus lateralis and vastus medialis muscles of the quadriceps femoris. Nine females reporting current signs and symptoms of PFP and 8 healthy females service as the control (CTL) group volunteered to participate in this study. Participants completed maximal and submaximal (25, 50, 75% MVC) isometric muscle actions of the quadriceps femoris at a leg flexion angle of 45 degrees below the horizontal plane of the lever arm. The involved limb for the PFP group and the dominant limb for the CTL group were selected for testing and all submaximal force levels were randomized. There was no (p > 0.05) group difference in EMG amplitude response for any muscle at any % MVC level. For the MMG amplitude, however, there was a main effect (p < 0.05) for group where the control group demonstrated greater MMG amplitude for each muscle. These findings suggest that the presence of PFP influenced mechanical aspect of muscle function as measured by MMG, but not the electrical properties (EMG). MMG may provide unique insight into the intrinsic effects on muscle function due to PFP. PMID- 19280796 TI - Blink reflex recovery in central and peripherally originated movement disorders of the cranio-cervical area: a comparative study. AB - BACKGROUND: Previous studies have shown that enhancement of blink reflex (BR) excitability exists in various movement disorders, notably those involving the cranio-cervical area. In this study the BR recovery curve of the late component (R2) and R2 area was evaluated to determine what changes occur under different conditions and the significance of these changes. METHODS: The recovery curve of the blink reflex (BR) was studied in patients with hemifacial spasm (HFS), post facial syndrome (PFS), blepharospasm (BS), segmental cranio-cervical dystonia (SD), cervical dystonia without BS (CD), and healthy control subjects. Following initial comparisons between six groups where the increased excitability observed in BS, SD and CD were assumed to be of central origin and that of HFS and PFS of peripheral origin; two additional groups were established and compared in terms of recovery times. RESULTS: Recovery values of 200, 400, 600, 800, 1000 ms and grades for all groups were significantly different when compared to the control group. The most significant difference observed between the groups was at 200 msn and the smallest at 800 msn interstimuli intervals. The subjects within the group referred to as the 'central origined group' recovered faster than those in the 'peripherally origined group' although no statistical difference was found between the two groups at stimulus interval. CONCLUSION: Our findings suggest that the excitability changes observed among the groups can be attributed to a number of factors. Although the nature and anatomical substrate of the primary pathology involving the reflex cycle leads - to some extent - to differently enhanced excitability patterns, such differences were not considered statistically significant. PMID- 19280797 TI - Cognitive impairment in encephalitis: P3 and MRI correlation. AB - OBJECTIVE: In view of paucity of studies on P3 in encephalitis, we report clinical, MRI and P3 changes in encephalitis patients. DESIGN: The diagnosis of encephalitis was based on clinical, MRI, ELISA and/or PCR and categorized into Japanese encephalitis (JE), dengue, herpes (HSE) and nonspecific group. Cognitive functions were evaluated 1 month after the illness when patient was able to cooperate for Mini Mental State examination (MMSE). P3 was carried out by odd ball auditory paradigm recording from Cz, Fz and Pz referred to linked mastoids. RESULTS: 30 encephalitis patients including 16 JE, 2 HSE and 12 nonspecific whose median age was 24 (13-53) years were prospectively evaluated. P3 was studied after a median duration of 3 months. MMSE was abnormal in 19 patients. CzP3 was abnormal in 6; 5 of whom had abnormal MMSE. MRI was abnormal in 18 showing thalamic lesion in 13, basal ganglia in 3 and cortical in 7 patients. MMSE was abnormal in all the patients with HSE, 81% with JE and 33% with nonspecific encephalitis. P3 was more frequently abnormal in patients with abnormal MMSE and MRI. CONCLUSION: Cognitive impairment is common in HSE and JE; abnormal MMSE and MRI are associated with P3 abnormality. PMID- 19280798 TI - An examination of the frequency-specific behavior of the mechanomyographic amplitude versus isometric torque relationship. AB - The purpose of this study was to examine the patterns of responses for mechanomyographic (MMG) amplitude versus isometric torque in different frequency bands for the vastus lateralis (VL), rectus femoris (RF), and vastus medialis (VM) muscles. Eleven men (mean +/- SD age = 20.1 +/- 1.1 yrs) performed submaximal to maximal isometric step muscle actions of the dominant leg extensors from 10% to 100% of the maximum voluntary contraction (MVC). During each muscle action, three separate surface mechanomyographic (MMG) signals were detected from the VL, RF and VM. Each MMG signal was decomposed into 9 different frequency bands (5-15, 15-25, 25-35, 35-45, 45-55, 55-65, 65-75, 75-85, and 85-95 Hz), and the root-mean-square amplitude of the signal in each frequency band was calculated. The results showed that for the VL and RF muscles, MMG amplitude plateaued from 80-100% MVC in the 15-25 and 25-35 Hz frequency bands. For the VM, however, the plateau in MMG amplitude from 80-100% MVC occurred in the 5-15 and 15-25 Hz bands. These findings indicated that there were both muscle- and frequency-specific discrepancies in the MMG amplitude versus isometric torque relationship that could be due to differences in muscle architecture and/or fiber type composition. PMID- 19280799 TI - Neuromuscular excitability changes in the vastus medialis following anterior cruciate ligament reconstruction. AB - PURPOSE: Quadriceps weakness following anterior cruciate ligament reconstruction (ACLR) is prevalent despite intensive rehabilitation. Diminished neuromuscular excitability is one potential factor that may limit muscular recovery following injury or surgery. The H-reflex provides a measure of alpha motorneuron (neuromuscular) excitability in the sensory-motor pathway of the respective muscle and nerve. To date the vastus medialis (VM) and soleus (SOL) H-reflexes have been examined primarily in control subjects with induced knee joint effusion. This prospective, randomized clinical trial evaluated the affect of ACLR, utilizing hamsting (HS) or bone-patellar tendon-bone (BTB) autograft, on VM and SOL H-reflex latency and amplitude in twenty subjects. METHODS: Preoperatively bilateral VM and SOL H-reflex tests were conducted. VM and SOL H reflexes were subsequently conducted on the involved lower extremity at 1 and 3 months post surgery. At each test session subjects completed visual analog scales and knee girth was measured. RESULTS: The VM H-reflex amplitude increased in the HS group at 3 months compared to 1-month post surgery (p<.05). Significant changes over time were also noted in the visual analog pain and functional scales and the mid-patella girth. CONCLUSIONS: The increased VM H-reflex amplitude at 3 months following HS autograft ACLR demonstrates an increase in VM neuromuscular excitability. Increased VM neuromuscular excitability was not evident in patients following BTB reconstruction. The increased neuromuscular excitability, observed only in the HS group, warrants consideration when selecting graft type for patients with extensive preoperative quadriceps dysfunction. PMID- 19280800 TI - The relationship between symptoms, clinical tests and nerve conduction study findings in carpal tunnel syndrome. AB - Carpal tunnel syndrome (CTS) is the most common entrapment neuropathy. The aim of the present study was to determine the relationship between patients reported symptoms and clinical tests with electrodiagnostic findings. Three hundred and nine patients with a mean age of 48.35 +/- 12.26 (range = 19-81 years) participated. Patients were assessed clinically and electrophysiologically. The main outcome measures were CTS related symptoms of pain and paraesthesia, Tinel sign, Phalen's test, distal sensory latency, and distal motor latency. The symptoms of presence of pain and diurnal paraesthesia showed a statistical relationship with the distal sensory latency. No relationship was found between the Tinel sign and either the distal sensory latency or the distal motor latency. Furtheremore, no relationship could be shown between Phalen 60 s, Phalen 45 s and distal sensory and motor latency. The Phalen 30 s had a significant relationship with both sensory and motor distal latency. The findings of this study indicate that both CTS related symptoms of pain, diurnal paraesthesia, and Phalen 30 s are associated with electrodiagnostic tests. PMID- 19280801 TI - Orbicularis oculi reflex abnormalities in patients with multiple sclerosis: a clinical, EMG, and MRI investigation. AB - Examination of the orbicularis oculi reflex (OOR) has been used for many years in the diagnostic algorithm of multiple sclerosis (MS) in a few aspects: to define existence and specific location of the lesions and to follow-up dynamic changes in pontine and supranuclear structures of the CNS. Correlation between MRI and electrophysiological data from OOR has not been widely studied in the patients with MS. The aim of the present study is to assess the changes of OOR in patients with MS and their relation to clinical and MRI brainstem findings. Forty-six patients were examined by complex EMG, MRI, and clinical protocol, aged from 21 to 55. Two groups were formed: first group of 40 patients with relapsing remitting MS (RRMS) and second group of six patients with secondary progressive (SPMS). About one third of the patients had neither clinical nor OOR or MRI brainstem abnormal changes. In 22.5% of the cases a clinical, EMG and MRI correlation, showing brainstem lesions, was found We observed different changes in OOR responses in about three fourths of all patients, respectively in 75% of patients with RRMS and in 83.3% of those with SPMS. OOR examination gives us evidence of additional brain lesions in 15% of our patients, whose MRI and clinical findings were unremarkable. Prolonged latency of the first and second component of OOR, additionally with asymmetry or lack of response, was more commonly related to the clinical manifestation. PMID- 19280802 TI - Complications of bariatric surgery: implications for the covering physician. AB - Bariatric surgery is the only effective option for sustained weight loss for morbidly obese patients. The increasing prevalence of obesity in America and the application of a laparoscopic approach to bariatric surgery have combined to dramatically increase the number of patients undergoing these types of operations. The number of bariatric surgeons and centers devoted to surgery of the morbidly obese is also rising. These facts lead to the assumption that there will be more patients with complications specific to bariatric surgery that must be cared for by general surgeons in the immediate future. Covering surgeons and those without expertise in bariatric surgery need to know how to diagnose and manage these potential complications in emergent and outpatient settings. This paper reviews some of the more common bariatric operations, complications, and conservative treatment options. PMID- 19280803 TI - The University of Cincinnati College of Medicine Department of Surgery. AB - The University of Cincinnati College of Medicine Department of Surgery has a legacy of discovery, transformation, and education that began more than 200 years ago. Through collaborations with related disciplines at the University of Cincinnati, industry, the government and military, as well as our partners in the community, we look forward to new innovations to continue the legacy and our ongoing mission to provide the best possible surgical care, research, and education of our future surgeon leaders. Our mission remains: 1) To provide comprehensive surgical services for Cincinnati and the surrounding region; 2) To train the next generation of surgeon leaders; 3) To advance the state-of-the-art and scientific basis of the discipline of surgery; and 4) To provide leadership in ensuring surgical health care for all members of the Cincinnati community. These principles have always been part of our legacy and ongoing challenges as an academic department of surgery. It is a proud tradition that we embrace and will carry into the future. PMID- 19280804 TI - Surgical considerations for the familial cancer syndrome, neurofibromatosis 1: a comprehensive review. AB - Neurofibromatosis 1 is one of the more common inheritable disorders that surgeons may encounter. A plethora of systemic associations, both benign and malignant, can affect these patients, and an acute awareness of these associations is essential for proper surgical care. A complete review of this disorder from the surgical perspective follows, highlighting the importance of this awareness. A brief review on the management and follow-up of surgical malignancies associated with this disorder is included. PMID- 19280805 TI - Laparoscopic gastrojejunostomy in patients with obstruction of the gastric outlet secondary to advanced malignancies. AB - Surgical palliation is an important therapeutic goal in patients with gastric outlet obstruction from cancer. The use of laparoscopic approaches for this condition has not been well studied. Our objective is to compare surgical outcomes of laparoscopic and open gastrojejunostomies in patients with gastric outlet obstruction secondary to advanced malignancies. We did a retrospective review of 20 patients who underwent a palliative gastrojejunostomy as their primary surgical procedure. There were 10 patients in the laparoscopic group and 10 patients in the open one. We identified no significant difference between groups in mean surgery time (116 vs 116 minutes) (P = 0.99), blood loss (23 vs 142 mL) (P = 0.19), or length of stay (8 vs 14 days) (P = 0.14). We also identified no difference in median time to tolerate a regular diet (7 vs 8 days) (P = 0.49) and median survival (11.2 vs 9.0 months) (P = 0.83). Delayed gastric emptying was the most common complication occurring in four patients. There is no detectable difference in surgical outcomes between laparoscopic and open gastrojejunostomies in the management of patients with obstruction of the gastric outlet secondary to cancer. Laparoscopic gastrojejunostomy is a safe and feasible operation in this setting. PMID- 19280806 TI - Computed tomography grading systems poorly predict the need for intervention after spleen and liver injuries. AB - Computed tomography (CT) grading systems are often used clinically to forecast the need for interventions after abdominal trauma with solid organ injuries. We compared spleen and liver CT grading methods to determine their utility in predicting the need for operative intervention or angiographic embolization. Abdominal CT scans of 300 patients with spleen injuries, liver injuries, or both were evaluated by five trauma faculty members blinded to clinical outcomes. Studies were graded by American Association for the Surgery of Trauma criteria, a novel splenic injury CT grading system, and a novel liver injury grading system. The sensitivity and specificity of each methodology in predicting the need for intervention were calculated. The kappa statistic was used to determine interrater variability. Twenty-one per cent (39/189) of patients with splenic injuries visible on CT scans required interventions, whereas 14 per cent (21/154) of patients with liver injuries visible on CT required interventions. The overall sensitivity of all grading systems in predicting the need for surgery or angioembolization of the spleen or liver was poor; the specificity seemed to be fairly good. When evaluators were compared, the strength of agreement for the various scoring systems was only moderate. Anatomic CT grading systems are ineffective screening tools for excluding the need for operation or embolization after splenic or hepatic trauma. Although insensitive, CT is a good predictor (highly specific) of the need for intervention if certain definitive abnormalities are identified. Considerable inconsistency exists in interpretation of abdominal CT scans after trauma, even among experienced clinicians. PMID- 19280807 TI - Effect of prior abdominal surgery, peritonitis, and adhesions on catheter function and long-term outcome on peritoneal dialysis. AB - Adhesions from previous surgery and peritonitis can lead to peritoneal dialysis catheter insertion failure, visceral injury, and poor dialysis function. Employing a laparoscopic approach, the effect of adhesiolysis on mechanical catheter complications and long-term catheter survival was prospectively examined in 436 catheter placement procedures having a 57.8 per cent prevalence rate of previous surgery. Adhesiolysis was required in 31.8 per cent of cases with prior surgery and in 3.3 per cent of procedures without previous operations. The incidence of adhesiolysis was directly related to the number of prior surgeries (P < 0.0001). The incidence of catheter insertion failure from extensive adhesions was 1.8 per cent. Survival probability free from catheter obstruction was lower in patients requiring adhesiolysis compared with subjects with prior surgery not requiring adhesiolysis (P = 0.01). Laparoscopic rescue procedures limited catheter losses from flow obstruction to only 0.7 per cent. As a result, long-term catheter survival was not different among patients regardless of prior surgery and/or adhesion status (P = 0.2). Scars on the abdomen and prior peritonitis do not predict the extent of adhesions and should not be used to judge eligibility for peritoneal dialysis. Presently, laparoscopy is the only practical way to provide optimal peritoneal access in patients with a history of surgery and peritonitis. PMID- 19280808 TI - Interim results of nutritional support through a button-type jejunostomy after total gastrectomy. AB - Nutritional deficiency is an inevitable consequence after total gastrectomy. We have recently developed a simple technique for nutritional support after total gastrectomy, in which the button jejunostomy is conducted on the afferent limb for long-term jejunal feeding. The purpose of this study was to evaluate the safety and efficacy of the low-profile button jejunostomy (LBJ) in patients with gastric malignancy. The records of 55 consecutive patients who were treated by total gastrectomy along with LBJ-making for gastric malignancy were prospectively reviewed and analyzed. Outcome parameters such as body weight, per cent ideal body weight (IBW), and nutritional parameters were regularly measured preoperatively and postoperatively. There were no complications related to LBJ. Among the patients, 53 who had survived at least 6 months after surgery were evaluated. The median caloric supplement by enteral feeding at home was 800 kcal per day. The median weight loss was 10.7 per cent of initial body weight. More than half of the patients achieved IBW and 40 patients (75%) were classified as low or no risk of malnutrition. This technique is easy to perform with minimal discomfort, and we believe that LBJ is useful for supporting dietary intake and for preventing malnutrition after total gastrectomy. PMID- 19280809 TI - Open thoracic window: a useful alternative for retained infected pleural collections in critically ill trauma patients. AB - Historically, tube thoracostomy, image-guided drainage, or an open thoracotomy has been indicated as the standard procedure for the management of patients with retained infected pleural collections (RIPC). These infections can be a debilitating and potentially lethal complication in already critically ill trauma patients. The purpose of this review was to evaluate the usefulness of an open thoracic window (OTW) as definitive therapy for the management of RIPC refractory to conventional therapies. The medical records of patients who underwent an OTW for RIPC were reviewed for the following: demographic data, primary diagnosis, clinical findings that explained the failure of the conventional management, bacteriology of the retained collection, and final outcome. Over a 3-year period, eight critically ill trauma patients who had sustained multiple system trauma and developed a RIPC were identified (six males and two females; average age, 47 years). Of the eight patients identified, six collections were in the right and two in the left pleural cavity. Staphylococcus aureus and Acinetobacter were the two most common bacterial isolates from these collections. All patients had undergone multiple and unsuccessful drainage attempts by thoracostomy tubes. Additionally, two of the patients also underwent image-guided drainage procedures, which proved to be unsuccessful. After creation of the OTW, all patients had complete resolution of the RIPC, and all were discharged alive from the hospital. During outpatient follow up, the OTW was found to have completely healed and required no further surgical intervention. The creation of long-term pleural drainage, with an OTW, facilitates and expedites the resolution of persistent infected pleural collections by providing more efficient surgical drainage and mechanical debridement. Our experience also shows this uncommon operation to be an effective alternative when conventional measures have failed. PMID- 19280810 TI - The impact of open lung biopsy on diffuse pulmonary infiltrates in patients with AIDS. AB - The evidence concerning open lung biopsy (OLB) for diffuse pulmonary infiltrates in patients with AIDS is limited. This study retrospectively evaluated the diagnostic and therapeutic yields of OLB compared with bronchoscopy for patients with AIDS with diffuse pulmonary infiltrate treated in the National Taiwan University Hospital from 1997 to 2004. There were 15 and 46 patients enrolled in the OLB and bronchoscopic groups in this study, respectively. As compared with the bronchoscopic group, patients from the OLB group had a higher chance of acquiring a specific diagnosis and possible change of therapy (73 vs 32.6% and 60 vs 21.7%, respectively; P < 0.05). There were two (13.3%) patients with prolonged air leak (greater than 7 days) in the OLB group, whereas four patients (8.7%) had procedure-related complications in the bronchoscopic group. There was no procedure-related mortality in the OLB group. This study demonstrated that OLB can be safely performed in select patients and provide a superior diagnostic and therapeutic benefit compared with bronchoscopy for diffuse pulmonary infiltrates in patients with AIDS. This procedure should be performed early in the clinical course to avoid irreversible clinical deterioration of the patients with severe illness. PMID- 19280811 TI - Is survival reduced for patients with anal cancer requiring surgery after failure of radiation? Analysis from a population study over two decades. AB - Chemoradiotherapy is the standard treatment for anal cancer. Surgery is reserved for failure of therapy, but there are limited data examining outcomes after surgery. From a prospective population-based database on radiation and surgical therapy, we compare outcomes for patients with anal cancer undergoing rectal resection after radiation with patients undergoing radiation alone. Patients undergoing surgical resection of the rectum after initial radiation (SRT) for squamous cell carcinoma of the anus, anal canal, cloacogenic zone, and overlapping lesions of the rectum and anal canal from 1983 to 2002 were identified from the Surveillance, Epidemiology and End Results database. Patient and tumor characteristics of SRT were compared with those of patients who underwent radiation alone (RT). Survival was calculated by the Kaplan-Meier test. There were 1202 patients undergoing RT and 48 patients undergoing SRT. RT and SRT had similar median age, gender, and grade of tumor. SRT had more patients with regional stage of disease (66.7 vs 42.4%, P = 0.001). Mean survival for SRT was, however, similar to RT (103 vs 96 months, P = 0.8). For patients with localized stage, survival for SRT and RT was similar (105 vs 98 months, P = 0.7). For patients with regional stage, survival for SRT and RT was similar (95 vs 83 months, P = 0.6). The presence of regional disease appears to be associated with surgical resection after radiotherapy. Mean survival for such patients is comparable to that of patients undergoing radiation alone. Because radiation is combined with chemotherapy, this suggests that salvage surgery after failure of therapy results in outcomes comparable to combination therapy alone. PMID- 19280812 TI - The effect of abdominal binders on postoperative pulmonary function. AB - Abdominal binders are ordered by some surgeons postoperatively for patient comfort and to prevent wound complications. There has been some question as to the compressive effect that an abdominal binder has on pulmonary function. We prospectively randomized 54 patients undergoing a midline laparotomy incision to two groups: a "binder" group and a "no binder" group. Preoperative pulmonary function tests (vital capacity and incentive spirometry) were measured. Postoperatively, pulmonary function tests, pulse oximetry, oxygen requirement, pulmonary and wound complications, pain control, time to ambulation, and hospital length of stay were examined. Vital capacity as a per cent of preoperative values on postoperative Day 1 for the binder and nonbinder groups were 64.7 and 54.6 per cent, respectively, but this was not statistically significant. Average level of pain using the visual analog pain scale on postoperative Days 1 through 3 in the binder versus nonbinder groups was 4 versus 8, 3 vs 6, and 3 versus 7, respectively. Time to ambulation was 18.6 hours in the binder group and 16.7 hours in the nonbinder group. Hospital length of stay in the binder and nonbinder groups was 3.9 days and 3.7 days, respectively. We conclude that abdominal binders in our patients with midline abdominal incisions had no significant effect on postoperative pulmonary function, but seemed to help with pain control. PMID- 19280813 TI - Preformed silastic silos in the management of necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is sometimes complicated by abdominal compartment syndrome, a clinical syndrome characterized by multiple organ dysfunction that arises as a consequence of increased intra-abdominal pressure. The evolving clinical picture of NEC sometimes requires "second-look" operations done after initial abdominal exploration to more accurately gauge the optimal extent of surgery. Placing intestines in a preformed, spring-loaded, transparent Silastic silo, traditionally used in the staged treatment of gastroschisis, addresses both situations: decompression of the abdomen and allowing periodic inspection of the intestines. Standard silos were used in three infants with advanced (Bell Class 3) NEC without perforation before definitive surgery. Clinical indices and laboratory values were recorded during the patients' hospital courses. All three infants had extensive areas of intestinal ischemia and necrosis. FiO2, acidosis, and urinary output remained stable or improved in two patients. Silo placement corrected abdominal compartment syndrome in the third patient. Intestinal resection was required in all infants, each achieving surgical resolution of NEC. Two patients ultimately died from respiratory and neurologic complications. Application of a silo addresses abdominal compartment syndrome as a complication of NEC and allows continual inspection of the intestines. Physiological indices may improve the patient's overall clinical status. PMID- 19280814 TI - The modified Fontan procedure and prolonged pleural effusions. AB - The modified Fontan procedure may be complicated by prolonged pleural drainage. Predisposing factors are not fully understood. This study examines perioperative variables associated with prolonged effusions. We examined the Fontan procedure in 41 patients. Mean age was 45 months (range, 9 to 113 months). Mean weight was 14.7 kg (range, 6.9 to 30.4 kg). Diagnoses included tricuspid atresia in 29 per cent, pulmonary atresia in 12 per cent, and double-outlet right ventricle in 12 per cent. Fontan revisions were excluded. The extracardiac Fontan was performed in 22 patients (54%) and the lateral tunnel (LT) in 18 (44%). Fenestration was performed in 56 per cent. Ten patients (24%) had prolonged pleural effusions. Mean chest tube output (24.2 vs 14.1 mL/kg/d, P < 0.01), days with chest tubes (10.2 vs 5.8 days, P < 0.01), and length of stay (20.8 vs 8.13 days, P < 0.01) were significantly greater in those with effusions. Preoperative mean pulmonary artery pressure was higher in patients with prolonged effusions (12.7 vs 9.90 mmHg, P = 0.001). No other factors were identified as risk factors. Prolonged pleural effusions (greater than 14 days) are common after the modified Fontan procedure. A higher preoperative mean pulmonary artery pressure may be predictive of these effusions. Fenestration, type of Fontan reconstruction, and size of extracardiac conduit did not predispose to postoperative effusions. PMID- 19280815 TI - Laparoscopic diagnosis and management of jejunal perforation resulting from accidental toothpick ingestion. PMID- 19280816 TI - An unusual case of intensive care unit ileus: a case of a posttraumatic pericecal herniation. PMID- 19280817 TI - Shotgun pellet embolus to the right heart from forearm entrance. PMID- 19280818 TI - Successful early fascial closure of an open abdomen during pregnancy. PMID- 19280819 TI - Endoscopic management of iatrogenic colon perforation during colonoscopy. PMID- 19280820 TI - A complex renal artery aneurysm successfully treated with ex vivo polytetrafluoroethylene tube graft reconstruction. PMID- 19280821 TI - Complete small bowel obstruction secondary to internal transomental herniation: report of an unusual 'swiss cheese' omentum. PMID- 19280822 TI - Intestinal occlusion resulting from a small bowel adenocarcinoma as first onset of Crohn's disease in a healthy patient. PMID- 19280823 TI - The torch is passed. Building on a solid foundation. PMID- 19280824 TI - Federal Trade Commission Red Flag Rules raise compliance concerns for dentists. PMID- 19280825 TI - 2009 NYSDA President Michael R. Breault. Consensus personality ready to lead Association in transition. PMID- 19280826 TI - Dental health certificate program elicits more questions and answers. PMID- 19280827 TI - Rationale and technique for achieving occlusal harmony. AB - For a patient to function normally with natural or restored dentition, the occlusal contacts should be in harmony with the musculature and the temporomandibular joint. Many times, this harmonious relationship is altered by restorative treatment, periodontal disease or tooth loss. If the adaptive capacity of the patient is less than the forces placed upon the tooth or restoration, failure in one form or another often results. This article presents a rational method the dentist can use to recognize and understand the markings made with articulating paper so that harmonious contacts can be detected and contacts that disturb occlusal harmony eliminated. PMID- 19280828 TI - Is bronchial asthma a risk factor for gingival diseases? A control study. AB - Asthma is a serious global health problem. People of all ages in countries throughout the world are affected by this chronic airway disorder, which can be severe and sometimes fatal. The prevalence of asthma is increasing everywhere, especially among children. Several oral health conditions are documented among asthmatic patients, such as an increased rate of caries development and reduced salivary flow; an increased prevalence of oral mucosal changes, like oropharyngeal candidiasis and gingivitis; and orofacial abnormalities. The study presented here was conducted to find a relationship between increased levels of plaque and gingivitis and bronchial asthma. Around 80 asthmatic patients were examined for their plaque and gingival status. Their scores were compared with a control group matched for age, sex and socioeconomic status. Results showed a significant increase in plaque and gingival scores among asthmatics as compared to the control group. Hence, there is a need to educate this group of patients about their increased risk of gingival disease and the importance of proper plaque control. PMID- 19280829 TI - Early orthodontic diagnosis and correction of transverse skeletal problems. AB - Rapid maxillary expansion (RME) is an orthodontic treatment commonly used in children to correct skeletal transverse problems of the maxilla. This clinical report introduces an effective way to diagnose and treat skeletal transverse problems achieving an orthopedic response, thereby reducing dental tipping and the need to overcorrect. Two clinical cases are used to illustrate an accurate skeletal diagnosis of the transverse dimension, orthopedic treatment using RME and a rationale for early orthodontic intervention. PMID- 19280830 TI - A multi-disciplinary approach to congenitally missing anterior teeth. AB - Congenitally missing teeth are not an unusual occurrence. What makes this case study unique is the combination of restorative challenges offered. The patient presented with the permanent upper right cuspid and lower anterior right and left central incisors congenitally missing, and with an underdeveloped upper right lateral incisor. Distinctive existing occlusion and tooth alignment difficulties precluded a simple approach of merely extracting the residual primary teeth and replacing the missing teeth prosthetically. Using orthodontics to create ideal interocclusal and interarch space was critical in preparing this case for prosthetic restoration. PMID- 19280831 TI - Treatment of gingival recessions by guided tissue regeneration and coronally advanced flap. AB - Gingival recession refers to the denudation of root surface caused by apical migration of the gingival margin as a result of destruction of the covering gingival tissue of the affected area. It is among the most frequent problems presented by periodontal patients and may have different etiologies and sequels. So far, several techniques have been devised and tested to treat gingival recession. The aim of this study was to assess the effectiveness of using a GTR resorbable collagen membrane in conjunction with coronally advanced flap (CAF) as compared to CAF alone in the treatment of Miller's Class I & II gingival recessions. Seven patients took part in the study, each providing either two or four facial recessions of 3 mm. to 6 mm., totaling 11 pairs of gingival recessions. The two paired sites within each patient were randomly assigned to one of the two treatments mentioned above. Prior to and six months after treatments, the following clinical parameters were measured and recorded: recession depth; probing pocket depth; clinical attachment level; width of keratinized gingiva; and width of recession. After six months, recession depth showed a mean reduction of 67.88% and 57.42% in the "GTR + CAF" and "CAF alone" groups, respectively. The mean difference between the groups was 1+/-0.33 mm (P=0.03). The results of this study indicate that Miller's Class I & II gingival recessions are amenable to treatment using the GTR technique with satisfactory outcome. PMID- 19280832 TI - Prosthetic treatment for severe open bite malocclusion and loss of several teeth. AB - This report describes a prosthetic approach that uses fixed partial dentures to establish stable posterior occlusion. Four fixed partial dentures were made of a silver-palladium-copper-gold alloy; the pontics were veneered with a light-curing indirect composite material. They were prepared using a thiouracil metal conditioner and seated with an adhesive luting cement. The clinical follow-up indicated that the prosthetic procedure reported here is applicable for severe anterior and posterior open bite malocclusion. PMID- 19280833 TI - ["Aut Viem Inveniam, aut facem". The need to interrelate the scientific periodic magazines in Mexico]. PMID- 19280834 TI - [Order of appearance of ossification centers in the elbow of Mexican children between 6 months and 12 years old]. AB - INTRODUCTION: Existing data of the order of appearance of ossification centers is from Anglo-Saxon literature. This literature describes the following order of appearance of the ossification centers of the distal humerus as follows: lateral condyle appears before one year of age, but it might delay up to 18-24 months; medial epicondyle between 5 to 6 years of age; the medal condyle between 9 to 10 years of age; the lateral epicondyle is the last one to ossify, between 10 to 12 years of age. Proximal radius ossifies between 3.8 and 4.5 years; olecranon at 6 to 8 years of age. METHODS: We studied 240 elbows and classified them in 6 groups according to the age of appearance of ossification centers. Anteroposterior and lateral radiographs were taken, and analyzed. RESULTS: We found a delay in the manifestation of the olecranon and the medial epicondyle. In base of our clinical observations, the ossification centers present later in Mexican children. PMID- 19280835 TI - [Survival of the T28 femoral stem. Comparison of polished versus roughed-surface finish]. AB - INTRODUCTION: Femoral stem loosening in total hip arthroplasty has a multifactorial etiology. T28 femoral stems have been made both roughed and polished-finish types, and differences in design are of interest in their effect on survival. MATERIALS AND METHODS: One hundred and sixty-seven stems (84 polished and 83 roughed-finish) placed between 1975 and 1982 were evaluated. Gruen zones were used to determine loosening and survival was determined by the Kaplan-Meier method. RESULTS: Revision was made in 24.8% of the cases. Twenty year survival rates were of 85.2% for roughed-finish stems and of 64.2% for polished-finish stems. CONCLUSIONS: We were able to confirm our hypothesis concerning the longer survival rate of the roughed-finish implant which was of 87.95% at 17 years of follow-up. The survival-time of the implant was independent of diagnosis, cementation, age and positioning. The survival-time of the T28 cemented implant does not only depend on its texture but it has a multifactorial origin. PMID- 19280836 TI - [Bone tumor frequency in adults and elderly]. AB - INTRODUCTION: We consider of great importance knowing the types of bone tumors and tumor-like lesions in adults and the elderly, which are increasing in proportion in our country. OBJECTIVES: (1) To analyze the frequency of bone tumors and tumor-like lesions in adults and elderly; (2) to determine any difference between these two groups; (3) and to analyze the most frequent sites of involvement. MATERIAL AND METHODS: We conducted a cross-sectional descriptive study that consisted in a retrolective review of all cases classified as a bone tumor or a tumorlike condition from year 2000-2006. We included all patients that presented with a bone tumor or tumor-like condition above 40 years-old. RESULTS: The most frequent diagnoses in patients between 40 and 60 years-old were plasmocytoma/myeloma (19.4%); giant cell tumor (14.1%) and metastases (12.3%). In patients older than 60 years, prevalence of bone tumor and tumor-like lesion was respectively: metastases (37.2%); plasmocytoma/myeloma (11.8%) and malignant fibrous histiocytoma (6.7%). Several differences were observed between these two groups. PMID- 19280837 TI - [Basic physical principles of ultrasonography, anatomy of the musculoskeletal system and ecographic artifacts]. AB - Ultrasound is a simple, non invasive and accessible imaging technique that allows the evaluation of the musculoskeletal system in real time, with the advantage of examining the joints in a dynamic way and with the aid of Doppler power, local inflammatory phenomena can be evaluated, without submitting the patient to radiation. It has the disadvantage of being operator dependant, which has to have a wide knowledge of the physical principles of ultrasonography, has to be familiar with the sonographic anatomy and musculoskeletal anatomy, but even more it is important that he has the ability to recognize the sonographic phenomena that can present as decoys, nominated artifacts. This knowledge is desirable to the specialist in musculoskeletal conditions, so that ecographic studies can be interpreted and recognizing artifacts and avoiding wrong diagnosis and unnecessary procedures. In the present review, the basic principles of ecography are analyzed, the normal sonographic anatomy of the musculoskeletal system ant the more common artifacts encountered in ecography. PMID- 19280838 TI - [Neurologic changes in patients with traumatic cord lesion based in the analysis of the sequential clinical history]. AB - OBJECTIVE: To determine changes in neurological status in patients with traumatic cord lesions before and after surgical treatment. MATERIAL AND METHODS: We conducted a descriptive cohort study, and measured 61 points of neurologic evaluation (sensory, motor, reflexes) of the Sequential Clinical History. RESULTS: We studied 28 patients with complete cord lesion (ASIA A) and 16 with incomplete cord lesions (9 ASIA B, 4 ASIA C, and 3 ASIA D). In the patients classified as ASIA-A, twenty-six did not change, and 2 had improvement in one neurologic level (sensory). In ASIA-B patients, four remained with no changes, and in 5 the lesion improved. The four patients with type C lesion, improved. All patients with ASIA-D, improved to ASIA-E. CONCLUSION: In patients with complete cord lesions (ASIA-A) and the worst prognosis, we did not find improvement in neurologic lesion. In incomplete cord injuries, improvement was variable, and it was more prevalent in preoperative period. PMID- 19280839 TI - [Continuous skeletal traction in multifragmentary fractures of radius and ulna combined with cast]. AB - INTRODUCTION: Fractures of the distal third of the radius are frequently encountered in emergency services. Until recently considered as benign, however, they represent a group of numerous therapeutic problems, as debated in anatomical, radiological and clinical studies. OBJECTIVE: To emphasize the importance of recovering the radial distance and the articular congruence in fractures of radius treated with a cast. HYPOTHESIS: To demonstrate that ligamentotaxis obtained with the application of the Cole-Obletz technique is a good method for treatment of distal radius fractures that have a component of impaction, extra or intra-articular lesion, a combination of both. MATERIAL AND METHODS: A retrospective study was realized on 10 patients with fractures of the distal radius, between February 2005 and February 2006, including adult patients of both genders. All patients received the Cole-Obletz method and were evaluated by antero-posterior and lateral wrist simple X-rays. According to Frykman classification, we compaired results in the immediate post-reduction X-rays with the results obtained at one month of treatment. Three different radiographic measures were evaluated: radius distance, inclination and volar tilt of the radius. RESULTS: We evaluated a group of 10 patients (80% female), with mean age of 52 years-old (24-79). Thirty percent of the patients (30%) presented a type VIII fracture, fourty percent a type VII fracture, and one patient for types V, IV, and II fractures, respectively. Articular congruence was achieved in the 80% of patients until full consolidation, in 10% of the patients only a 50% of radius distance was completed. In a patient anatomic reduction was lost on day eight. DISCUSSION: Treatment of complex distal radius fractures with plates and screws for maintaining anatomic reduction are not applicable in our socioeconomic context. This study was aimed to prove that the Cole-Obletz method permits to maintain adequate reduction, it is reproducible, and is achievable at low costs. PMID- 19280840 TI - [Musculoskeletal tumor evaluation with 99mTc-Tetrofosmin]. AB - INTRODUCTION: (99m)Tc-tetrofosmin is an efficient agent as a tumor marker. Several studies have proven its efficiency in detection and localization of tumors of the breast, larynx, thyroid, parathyroid glands, lung, brain, skin, lymphatic and musculoskeletal tissues with a sensitivity and specificity of 95% to 100%. It is used as a predictor of therapy efficacy and to localize remaining and recurrent tissue as well as local and distant extension. OBJECTIVE: To determine the sensitivity, specificity and predictive values of the three-phase 99mTc-Tetrofosmin scan in detection of malignant lesions of the musculoskeletal tissue. METHODS: Seventy-three patients of both genders with suspected malignant musculoskeletal lesions were studied by three-phase (vascular, tissue and delayed images) 99mTc-tectrofosmin scans. RESULTS: One-hundred-percent sensitivity, 67.3% specificity, 55% positive predictive value, and 100% negative predictive value. CONCLUSION: Three-phase 99mTc-tectrofosmin scan is useful in detection of malignant lesions of the musculoskeletal system due to its high sensitivity and positive predictive value. Nevertheless, benign pathology such as active inflammation is a source of false positives and the attending physician must consider the aid of complementary studies such as histopathology. PMID- 19280841 TI - [Femur overgrowth as presentation of an osteoid osteoma]. AB - INTRODUCTION: Osteoid osteoma is a benign tumor that typically produces night pain that improves with salicilates. This condition, rarely affects children younger than 5 years-old. CASE REPORT: We present a 2-year-old boy with lower limb discrepancy due to overgrowth of the femur as first manifestation of an osteoid osteoma. The patient did not complain of pain. The tumor localization was intramedular, and diagnosis was confirmed by pathology after broad resection of the tumor. DISCUSSION: This case report shows an unusual presentation of an osteoid osteoma in a young patient, with lower limb discrepancy caused by overgrowth of the femur in absence of pain. PMID- 19280842 TI - [Arthroscopy of the trapeziometacarpal joint: anatomical study on cadavers]. AB - INTRODUCTION: Osteoarthritis of the trapeziometacarpal joint is one of the most common degenerative diseases of the hand for which a variety of surgical procedures have been developed. Arthroscopic techniques offer good results, although they have not been standardized. OBJECTIVE: To identify the main anatomical structures in relation to arthroscopy portals from the trapeziometacarpal joint and to determine their security area. MATERIAL AND METHODS: We conducted a transversal and descriptive study in which the anatomy was analyzed in seven cadaveric specimens. Fourteen thumbs were dissected after the arthroscopic portals were made. In all of the specimens the distance between the radial artery, the articular line and the arthroscopy portals was measured. RESULTS: The radial artery was found dorsal to the Extensor Pollicis Longus (EPL), 4-5 mm proximal to the dorsal arthroscopic portal, 4-8 mm to the articular line and 8-11 mm to the Extensor Pollicis Brevis (EPB). In two cases sensitive branches of the radial nerve were found through the dorsal portal. CONCLUSION: The anatomical evaluation defined a secure area for the dorsal portal between the EPL and the EPB. The proximal radial zone to the EPL must be avoided so that the dorsal branch of the radial nerve is not damaged. PMID- 19280843 TI - [Jean Lelievre (1914-1969)]. PMID- 19280844 TI - Shiga-toxin producing Escherichia coli and the hemolytic uremic syndrome: what have we learned in the past 25 years? PMID- 19280845 TI - Global epidemiology of pneumococcal disease--new prospects for vaccine control. PMID- 19280846 TI - Current concepts of the pathogenesis of RSV bronchiolitis. PMID- 19280847 TI - How can we eradicate pertussis. PMID- 19280848 TI - Recognition and management of encephalitis in children. PMID- 19280849 TI - Microbiology and management of pleural empyema. AB - Empyema is apparently becoming more common, with pneumococcus being the most common pathogen detected in Europe and the USA. However, group A streptococcus and S. aureus pneumonia are individually more likely to progress to empyema. Serotype 1 pneumococcus is frequently implicated and the reasons for an apparent increase in incidence remain unclear. Management requires antibiotics and removal of pus either by fibrinolysis or primary drainage. PMID- 19280850 TI - Pneumococcal conjugate vaccines probe studies: the solution points to the problem. PMID- 19280851 TI - New findings in primary immunodeficiency. PMID- 19280852 TI - Routine use of influenza vaccine. PMID- 19280853 TI - Challenges in the evaluation and management of bone and joint infections and the role of new antibiotics for gram positive infections. AB - CA-MRSA isolates are increasingly common pathogens in children with acute, hematogenously acquired osteomyelitis and clinicians have to consider this pathogen in the selection of initial empiric treatment. With the emergence of pvl+ CA-MRSA isolates, unusual manifestations of acute osteomyelitis have become common including venous thromboses and multiple sites of infection. MRI is currently the imaging modality of choice for osteomyelitis. Drainage of purulent collections is also more common in association with the pvl+ strains. The role of newer antibiotics such as linezolid and daptomycin for the treatment of CA-MRSA musculoskeletal infections in children needs further study. PMID- 19280854 TI - The significance of serotype replacement for pneumococcal disease and antibiotic resistance. PMID- 19280855 TI - Childhood intra-thoracic tuberculosis. PMID- 19280856 TI - The antibiotic pipeline in pediatrics. PMID- 19280857 TI - Paediatric drug development and clinical trials. PMID- 19280858 TI - Influenza in pregnancy: the case for prevention. PMID- 19280859 TI - Febrile neutropenia in children with cancer. AB - There is a need for increased consensus in the definition of fever and neutropenia, the approach to risk stratification (including outpatient therapy and early discharge) and choices of empiric antimicrobial therapy in children. There has been an increased incidence of Gram positive infection in FN patients, in particular with VGS in patient with AML. However, Gram negative bacteria are still responsible for most of the mortality associated with FN. Piperacillin/tazobactam, cefipime, or meropenem are all effective first-choice antimicrobial monotherapy in FN. There is no good evidence for adding an aminoglycoside compound to the initial empiric therapy regimen. Following local microbiological data is of utmost importance in choosing the right empiric antimicrobial regimen for a particular institution. Outpatient management of a well-defined subset of low-risk patient for bacterial invasive infection with intravenous ceftriaxone or oral ciprofloxacin and daily re-evaluation is possible. Early CT of the chest (after 5-7 days of FN) in high-risk patients is essential to make a prompt diagnosis of pulmonary aspergillosis and improve outcome. PMID- 19280860 TI - The PANDAS syndrome. PMID- 19280861 TI - Fever in the returned paediatric traveller. PMID- 19280862 TI - Molecular diagnostics of primary immunodeficiencies: benefits and future challenges. PMID- 19280863 TI - Human genetic resistance to malaria. AB - This brief chapter highlights the need for caution when designing and interpreting studies aimed at seeking new genes that may be associated with malaria protection, or investigating the potential mechanisms for protection in promising candidates. Judging genetic effects on the basis of the wrong clinical phenotype and missing true protective genes because their protective effects are masked by unpredictable epistatic effects are major potential pitfalls. These issues are by no means unique to malaria: in recent years, the importance of larger sample sizes and careful phenotypic definitions have become appreciated increasingly, particularly for genome-wide studies of complex diseases (Cordell and Clayton, 2005; Burton, Tobin and Hopper, 2005). Until recently, research in the field of malaria genetics has not enjoyed the sort of funding afforded to similar work investigating diseases of importance to the developed world. However, in the last few years, coupled with advances in genetic diagnostics that have led to massive automation and falling costs per gene explored, momentum has grown towards more generous funding that brings with it the opportunity for much larger, multisite cohesive studies. The stage is set for a giant leap forward in the coming years. PMID- 19280864 TI - Dystonia: a clinical approach. AB - Dystonia is a common movement disorder characterised by abnormal postures of the affected body part. It has a very varied presentation and numerous causes, and this can create difficulties with diagnosis and appropriate investigation. This article aims to provide a clinical approach to patients with dystonia, focussing on how to create a differential diagnosis and to plan rational testing. PMID- 19280865 TI - Prognostic value of electrocardiography and electroencephalography in patients with ischemic stroke. AB - PURPOSE: Stroke is the most common disease among neurological disorders, often resulting in handicap, complications and mortality. Recent studies revealed that electrocardiography (ECG) and electroencephalography (EEG) were noninvasive but effective methods to assess the prognosis of stroke patients. The purpose of this study is to evaluate the use of EEG and ECG for the prognosis of ischemic stroke. METHODS: Thirty five patients with supratentorial ischemic stroke and muscle weakness were included in the study carried out at the Ali-EbneAbitaleb Hospital, Rafsanjan, Iran. At admission, muscle power was recorded for every patient. EEG and ECG were performed on patients within 12 hours of admission. The muscle power was re-evaluated for all study cases at 3 months. The data were analyzed using SPSS software; Mann-Whitney and Kruskal-Wallis tests. RESULTS: Opium addiction (45.7%), hypertension (40%), and smoking (28.6%) were the most common risk factors. ECG change was seen in 71.4% of patients (ST-T change: 51.4%, QT: 22.9%). No significant correlation was seen between ECG and muscle power changes at three months among stroke patients. 57.1% of patients had mild to severe abnormality in EEG and a significant correlation was seen between EEG and muscle power changes at three months (P= 0.01). CONCLUSION: This study showed that EEG abnormality was positively correlated with poor prognosis in our patients with ischemic stroke. This study did not show any significance for ECG, which may be due to a small patient number, and further study is needed. PMID- 19280866 TI - Uncoupling of protein C and antithrombin III activity in cerebral ischemia patients associated with cutis marmorata. AB - PURPOSE: Cutis marmorata is a cutaneous livedoid disorder which can be differentiated from livedo reticularis in both clinical and pathological presentations. Unlike Sneddon syndrome, a detailed immunocoagulation profile has not yet been delineated for cutis marmorata in patients with cerebral ischemia. METHODS: To analyze the immunocoagulation profile in cutis marmorata patients associated with cerebral ischemia (CMCI) in a series of 135 cerebral ischemia patients. RESULTS: A total of 32 patients were found to have cutis marmorata. The blood protein C activity, protein S activity, antithrombin III activity, platelet count, fibrinogen and frequency of abnormal antiphospholipid antibody level were similar among 32 CMCI patients, 103 cerebral ischemia patients without cutis marmorata, and 35 healthy subjects. However, uncoupling of protein C and anti thrombin III was observed in CMCI patients. Serum antinuclear antibody and Venereal Disease Research Laboratory were not detected in these patients. CONCLUSION: Cutis marmorata is not uncommon in our ischemic stroke patient population, and is characterized by uncoupling of protein C and antithrombin III with altered thrombin hemostasis. Our findings raise the need for a careful cutaneous examination in patients with ischemic stroke. Abnormal immunocoagulating profile should alert physicians to the risk for cerebral ischemia even in the absence of other cardiovascular risk factors. PMID- 19280867 TI - Lateral medullary infarction presenting contralateral palatal paresis. AB - Palatopharyngeal paresis has never been reported to be contralateral in the lateral medullary infarction (LMI). A 65-year-old lady with acute dorsolateral infarction in the left medulla presented mild hoarseness, mild dysphagia, mild gait ataxia along with marked hypalgesia and thermal anesthesia on the right limbs. To our surprise, palatal weakness was on the right side, instead of being on the left side as expected in typical Wallenberg syndrome. The palatal paresis was noted during voluntary phonation but turned into normal movement while gag reflex was induced. An involvement of corticobulbar fibers before synapsing into the ipsilateral caudal medulla or a selective involvement of the peri-ambigual reticular formation and/or its post-synaptic connection fiber destined to the contralateral swallowing center presumably underlies such a rare contralateral palatal paresis. PMID- 19280868 TI - Dissection of the posterior inferior cerebellar artery in a young adult with cerebellar infarct. AB - The posterior inferior cerebellar artery (PICA) is frequently involved in dissection of the vertebral artery (VA); however, isolated PICA dissection has rarely been reported. A 37-year-old man experienced acute and progressive drowsiness, vertigo, occipital headache, vomiting, and ataxia. There was no precedent trauma or chiropractic manipulation. Neurologically, he had dysmetria of the left extremities. His NIHSS score was 3. Brain magnetic resonance imaging showed an acute cerebellar infarct in the left PICA territory. Magnetic resonance angiography showed a faint signal adjacent to the junction of the left VA and PICA, suggesting a vascular shadow. Catheter angiography showed focal stenosis with a post-stenotic fusiform aneurysmal dilatation of the left proximal PICA that was highly suggestive of dissection with pseudoaneurysm formation. He was treated with clopidogrel and was free of neurological symptoms 3 months after the stroke event. Isolated PICA dissection may be considered in patients with PICA territory infarct or subarachnoid hemorrhage. Treatment depends on the manifestations; ruptured dissecting aneurysms are often treated with surgery or embolization, and infarcts are usually treated with antithrombotic agents. PMID- 19280869 TI - Capsular warning syndrome: report of a case. AB - The "capsular warning syndrome" (CWS) is characterized by recurrent stereotypical episodes of motor and/or sensory dysfunction without cortical signs. CWS is a clinically well recognized entity, and carries a significant risk of capsular infarct. The ischemia is most likely ascribable to hemodynamic changes in diseased small penetrating vessels. Treatment remains controversial and none has been proven effective. We described a 66-year-old man having 15 episodes of stereotypical transient ischemic attack within four days. The findings of the diffusion-weighted image showed abnormalities confined concurrently to the left lateral thalamus, posterior globus pallidus, and posterior corona radiata while the internal capsule was spared. Theses findings suggest involvement of the territory of anterior choroidal artery. We also documented changes in the pattern of attacks after initiation of intravenous urokinase. PMID- 19280870 TI - Behcet's disease with involvement of major arteries: a case report. AB - Vascular involvement is not infrequent in Behcet's disease (BD). It is generally seen in the form of superficial thrombophlebitis or occlusion of major veins. In rare instances, arterial occlusion and aneurysm formation may be seen in BD. We reported a young male with BD, diagnosed at the age of twenty for relapsing and remitting oral ulceration, skin rash, arthralgia and ocular painful redness for three years. At the age of 21, he had recurrent abdominal aortic aneurysm and inconspicuous neurological manifestations including dizziness, tinnitus and transients of blurred vision. The carotid angiography disclosed the occlusion of bilateral common carotid arteries (CCA). A carotid endarterectomy was subsequently performed to reduce the risk of stroke. The pathological examination of the occluded segment of CCA revealed chronic inflammation, which was attributable to BD. There was no atherosclerotic change. To the best of our knowledge, this is the first case report of concurrent bilateral CCA occlusion and relapsing abdominal aortic aneurysm. Even in the absence of specific neurological symptoms, we suggest that cerebrovascular investigation need to take into consideration in BD patients with unexplained cranial symptoms. PMID- 19280871 TI - Reversible posterior leukoencephalopathy syndrome caused by blood transfusion: a case report. AB - This is a case report of a 32-year-old woman with chronic severe anemia who developed headaches and seizures 5 days after receiving a blood transfusion of eight units (1600 ml) of packed red blood cells. Magnetic resonance imaging indicated vasogenic edematous lesions bilaterally over the occipital lobes that were consistent with reversible posterior leukoencephalopathy syndrome (RPLS). Her blood pressure was normal, and no other contributing factors for RPLS were found. It is likely that the initiator was the large volume of transfused blood, which disrupted cerebral autoregulation and damaged the vasculoendothelial system. Similar cases of RPLS following transfusion have been reported, and all reports involved middle-aged females with chronic severe anemia who received large volumes of transfused blood within a short period of time. Although blood transfusion is a common procedure with rare neurological complications, great caution should be taken with chronic severely anemic patients because a rapid elevation in hemoglobin may precipitate RPLS. PMID- 19280872 TI - Very early hemorrhagic transformation of a subcortical infarction. AB - Hemorrhagic transformation (HT) of an ischemic stroke within 6 hours has never been documented. We reported a case of 65-year-old female experiencing sudden onset of slurred speech and right side weakness. Her past history disclosed rheumatic heart disease and atrial fibrillation. The National Institutes of Health stroke scale (NIHSS) score was 20. The brain computed tomography one hour after symptom onset revealed a faint hypodense lesion in the left striatum. The lesion, however, was transformed spontaneously into a large hematoma within 3 hours of symptom onset. This case thus developed a very early HT of a subcortical infarct of possible cardioembolic origin and high initial NIHSS score. Though rare, very early spontaneous HT does happen. PMID- 19280873 TI - [Topographical disorientation: viewpoints from behavioral neurology]. AB - To navigate freely and safely in familiar environments is essential for humans in everyday life. This ability, however, can be deprived of after a focal cerebral damage or degenerative disease, such as Alzheimer's disease. In this article, the author introduces the concept of way finding behavior from animals to humans, and its dysfunction or topographical disorientation (TD) in humans which develops after focal cerebral damage and dementing illnesses. The author also links the neuro-anatomical structures, the cognitive functions related to way finding and the clinical manifestations of TD, with an attempt to clarify the brain behavioral relationship. For neurologists, the awareness of TD and its knowledge is helpful to make a correct diagnosis and to improve the quality of life of their patients. PMID- 19280874 TI - [Guidelines for the general management of patients with acute ischemic stroke]. AB - The content of the second edition of "Guideline for General Management of Patients with Acute Ischemic Stroke" was amended from the first edition of that of the Taiwan Stroke Society in 2002. The format of the guideline followed the common unified instruction for the project of "The establishment of clinical guidelines for the top 10 payments diseases of the National Health Insurance at the departments of inpatients, emergency and outpatients" as recommended by the National Health Research Institutes (NHRI). The guideline was revised after several official meetings of local experts, as well as citation from the latest updated guidelines of the United States and the European Stroke academic groups. Before editing notice, the final evaluation was performed by the review team of the NHRI. Application of the guideline is dedicated or designated to the patients with acute ischemic stroke, and which is applied only limited to the general management. Guidelines for subacute or chronic phase, or the specific treatment for ischemic stroke patients will be published in separated articles. Management of most of the needs for patients with acute ischemic stroke must be completed in a very short period of time. It is recommended that hospitals providing stroke service to set up stroke unit, and to organize an integrated stroke team consisting of specialists from multiple disciplines. Upon arrival to the hospitals, patients should undergo the brain computed tomography, and related examinations and assessment as soon as possible to guide the choice of treatment reference for acute intervention. Intravenous recombinant tissue plasminogen activator treatment within three hours is effective in reducing disability for patients with acute ischemic stroke. Ischemic stroke patients with or without persistent symptoms should start antiplatelet therapy immediately, generally aspirin. Dose-adjusted warfarin (INR range of 2.0-3.0) is recommended for ischemic stroke patients with persistent or paroxysmal atrial fibrillation to prevent secondary embolism. The routine use ofheparin and drugs theoretically preventing further brain injury, including steroids, neuroprotectants, plasma volume expanders, barbiturates, and streptokinase, has not been proven benefits for recommendation. PMID- 19280875 TI - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy. PMID- 19280876 TI - Effects of alendronate and alfacalcidol on the femoral bone mass and bone strength in orchidectomized rats. AB - The purpose of the present study was to compare the effects of alendronate (ALN) and alfacalcidol (ALF) on the femoral bone mass and bone strength in orchidectomized rats and to clarify the skeletal benefits of combined administration of ALN and ALF. Fifty male Sprague-Dawley rats, 3 months of age, were randomized by the stratified weight method into five groups: the age-matched control (CON), orchidectomy (ORX), ORX + ALN (2.5 microg/kg, s.c., 5 times a week), ORX + ALF (0.1 microg/kg, p.o., 5 times a week), and ORX + ALN + ALF groups. After 12 weeks of feeding, the femoral distal metaphysis and mid diaphysis were processed for peripheral quantitative tomographic analysis and biomechanical testing. In the femoral distal metaphysis, ALN prevented the ORX induced reduction in the trabecular volumetric bone mineral density (vBMD) and breaking energy, and ALF prevented the ORX-induced reduction in the trabecular vBMD and increased the breaking energy to values above those observed in the CON group. Both ALN and ALF increased the maximum load to values above those observed in the ORX group. The improvements in parameters described above were more pronounced when ALN and ALF were administered in combination. In the femoral mid diaphysis, on the other hand, ALN did not significantly affect the cortical bone parameters, whereas ALF increased the cortical area and maximum load to values above those observed in the ORX group. Furthermore, no apparent benefit of combined administration of ALN and ALF was observed. These findings suggest differential effects of ALN and ALF on femoral bone mass and the beneficial effects of combined administration of ALN and ALF on the trabecular bone of the femur in ORX rats. PMID- 19280877 TI - Alteration of serotonin 2C receptor expression in the aorta and the pulmonary artery in rats exposed to hypoxia. AB - In the present study we reported the expression of the 5-HT2C receptor (5-HT2C-R) in the aorta and the pulmonary artery in rats exposed to hypoxia, and examined its role in vascular smooth muscle cells. Immunochemistry, quantitative RT-PCR, and western blot were used to examine 5-HT2C-R expression. Small interfering RNA (siRNA) was used to explore the role of 5-HT2C-R in vascular smooth muscle cells proliferation and contraction induced by 5-HT.5-HT2C-R immunoreactivity was confined predominantly to the plasma membrane in the rat aortic smooth muscle cells (ASMCs) and the pulmonary arterial smooth muscle cells (PASMCs). 5-HT2C-R mRNA expression in the aorta of hypoxic pulmonary hypertension (HPH) rats was more than that in the pulmonary artery. Hypoxia-induced 5-HT2C-R protein expression in the aorta was less than that in the pulmonary artery, indicating that 5-HT2C-R might mediate different responses of the aorta and the pulmonary artery to 5-HT in HPH. We also demonstrated that RNA editing may not be involved in the different expression levels of 5-HT2C-R in the aorta and the pulmonary artery. The higher expression of 5-HT2C-R in the pulmonary artery versus the aorta of HPH rats was partly due to the increased protein stability. siRNA was used to knock down the expression of 5-HT2C-R. 5-HT2C-R-specific siRNA reduced the proliferation and contraction of vascular smooth muscle cells induced by 5 HT. In conclusion, the different expression patterns of 5-HT2C-R protein in the aorta and the pulmonary artery suggest that this 5-HT receptor may be involved, at least partly, in modulating different responses of systemic vessels and pulmonary vasculature to 5-HT in HPH. PMID- 19280878 TI - Reversion of P-glycoprotein-mediated multidrug resistance in human leukemic cell line by carnosic acid. AB - One of the common hindrances to successful chemotherapy is the development of multidrug resistance (MDR) by tumor cells to multiple chemotherapeutic agents. In this regard, P-glycoprotein (P-gp) acts as an energized drug pump that reduces the intracellular concentration of drugs, even of structurally unrelated ones. The modulators of P-gp function can restore the sensitivity of MDR cells to anticancer drugs. Therefore, to develop effective drug-resistance-reversing agents, we evaluated the P-gp modulating potential of carnosic acid (CA) in multidrug-resistant K562/AO2 cells in the present study. The reversing effect of CA was evaluated by determining the inhibition rates of cell viability with 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assays. The intracellular adriamycin fluorescence intensity and the expression of P-gp were measured by flow cytometry (FCM). Meanwhile, the subcellular distribution of adriamycin was detected via Laser Scanning Confocal Microscopy (LSCM). The mRNA expression of mdrlwas then detected via semiquantitative reverse transcription polymerase chain reaction (RT-PCR). The findings showed that CA decreased apparently the Inhibition Concentration 50% (IC50) of adriamycin by increasing its intracellular concentration and thus enhancing the sensitivity of K562/AO2 cells. Adriamycin was distributed evenly in the cytoplasm when the cells were treated with CA. The expression of mdrl was decreased. Overall, the results indicated that CA can serve as a novel, non-toxic modulator of MDR, and it can reverse the MDR of K562/AO2 cells in vitro by increasing intracellular adriamycin concentration, down-regulating the expression of mdrl, and inhibiting the function of P-gp. PMID- 19280879 TI - Down-regulation of the expression of angiotensin II type 1 receptor in neonatal rat cardiac fibroblast by activation of PPARgamma signal pathway. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) is one of the hormone nuclear receptors. Recent data have shown that activation of PPARgamma signal pathway has many positive effects on cardiovascular system. The goals of this study were to determine whether PPARgamma activator affects cardiac fibrosis and the possible mechanisms. Cardiac fibroblasts (CFs) of SD neonate rats were used in the study. Cells were divided into 4 groups: I--control group; II- pioglitazone group (Piog--PPARgamma agonist); III--angiotensin II (Ang II) group; and IV--Piog + Ang II group (Piog plus angiotensin II). mRNA and protein expression of collagen type I, III and angiotensin II type 1 receptor (AT1-R) were tested by reverse transcription--polymerase chain reaction and Western blotting. With the inhibition of actinomycin D, we investigated the impacts of Piog on the stability of AT1-RmRNA. Compared with group I, the mRNA and protein expression of collagen type I, III and AT1-R were up-regulated in group III (P < 0.05). However with the effects of Piog in group IV, the expressions mentioned above were attenuated significantly (P < 0.05). With the effects of actinomycin D, AT1-RmRNA was reduced at the same degree in control and Piog groups at the same time points. These results indicated that treatment with Piog can attenuate Ang II-induced collagen synthesis in CFs through down-regulation of the AT1-R expression. With the intervention of actinomycin D, we suggested that PPARgamma agonist didn't affect the stability of AT1-RmRNA. PMID- 19280880 TI - Attenuation of lung inflammation and pro-inflammatory cytokine production by resveratrol following trauma-hemorrhage. AB - Although studies have demonstrated that resveratrol administration following adverse circulatory conditions is known to be protective, the mechanism by which resveratrol produces the salutary effects remains unknown. We hypothesized that resveratrol administration in males following trauma-hemorrhage decreases cytokine production and protects against lung injury. Male Sprague-Dawley rats underwent trauma-hemorrhage (mean blood pressure 40 mmHg for 90 min, then resuscitation). A single dose of resveratrol (30 mg/kg of body weight) or vehicle was administered intravenously during resuscitation. Twenty-four hours thereafter, tissue myeloperoxidase activity (a marker of neutrophil sequestration), cytokine-induced neutrophil chemoattractant (CINC)-1, CINC-3, intercellular adhesion molecule (ICAM)-1, and interleukin (IL)-6 levels in the lung and protein concentrations in bronchoalveolar lavage fluid were measured (n = 6 rats/group). One-way ANOVA and Tukey's test were used for statistical analysis. Trauma-hemorrhage increased lung myeloperoxidase activity, CINC-1, CINC 3, ICAM-1, and IL-6 levels and protein concentrations in bronchoalveolar lavage fluid. These parameters were significantly improved in the resveratrol-treated rats subjected to trauma-hemorrhage. The salutary effects of resveratrol administration on attenuation of lung injury following trauma-hemorrhage are likely due to reduction of pro-inflammatory mediators. PMID- 19280881 TI - Characterization of a stable steroidogenic caprine luteal cell line transformed by a temperature-sensitive simian virus 40. AB - A caprine luteal cell line (tsCLC-D) that synthesizes progesterone (P4) was established following by transformation with a temperature-sensitive A209 (tsA209) mutant of simian virus 40 (SV40). The transformed cells have temperature sensitive for morphology, cell propagation and progesterone steroidogenesis. At the permissive temperature of 34 degrees C, these cells were spindle-shaped and grew with a similar rapidity as tumor cells. However, at the nonpermissive temperature of 40 degrees C, the cells have exhibited a round shape and ceased to proliferate because the gene for maintenance of transformation was not expressed. The tsCLC-D cell line responds to 8-Br-cyclic AMP, 22-hydroxycholesterol and pregnenolone treatment with an increase in progesterone biosynthesis. This cell line still express StAR protein, 3beta-HSD and P450scc enzyme of three kinds of steroidogenic protein and enzymes, this characteristic is similar to normal luteal cell. However, the addition of any doses oLH did not increase progesterone secretion. We speculate that tsCLC-D might lose the responsiveness to gonadotropins during the immortalization process, while retaining steroidogenic enzyme activity and progesterone production. To our knowledge, this is the first report of a stable cell line derived from corpus luteum of ruminant. The tsCLC-D retains steroidogenic capacity, which will make this cell line useful for the studies of regulation of steroidogenesis. PMID- 19280882 TI - Synergic co-activation of muscles in elbow flexion via fractional Brownian motion. AB - In reflex and volitional actions, co-activations of agonist and antagonist muscles are believed to be present. Recent studies indicate that such co activations can be either synergic or dyssynergic. The aim of this paper is to investigate if the co-activations of biceps brachii, brachialis, and triceps brachii during volitional elbow flexion are in the synergic or dyssynergic state. In this study, two groups with each containing six healthy male volunteers participated. Each person of the first group performed 30 trials of volitional elbow flexion while each of the second group performed 30 trials of passive elbow flexion as control experiments. Based on the model of fractional Brownian motion, the intensity and frequency information of the surface electromyograms (EMGs) could be extracted simultaneously. No statistically significant changes were found in the control group. As to the other group, results indicated that the surface EMGs of all five muscle groups were temporally synchronized in frequencies with persistent intensities during each elbow flexion. In addition, the mean values of fractal dimensions for rest and volitional flexion states revealed significant differences with P < 0.01. The obtained positive results suggest that these muscle groups work together synergically to facilitate elbow flexion during the co-activations. PMID- 19280883 TI - Lower side effects of milnacipran than paroxetine in the treatment of major depression disorder among Han Chinese in Taiwan. AB - Milnacipran is a dual-action antidepressant which inhibits both serotonin and norepinephrine reuptake. To our knowledge, it has limited affinity for most monoamine neurotransmitter receptors. With limited pharmacokinetic interaction with the cytochrome 450 system, milnacipran may have a low risk in drug interaction. The present study compares milnacipran with paroxetine, a selective serotonin reuptake inhibitor (SSRI) and which has been used clinically for years to evaluate the efficacy, patient tolerance, and side effects in the treatment of major depression. The study took place in two medical centers located in Northern and Southern Taiwan. Six-three participant who met the Diagnostic and Statistical Manual of Mental Disorder 4th edition (DSM-IV) criteria for a major depressive disorder and a total Hamilton Depression Rating Scale (HAM-D) score > or = 16 on the 17 item scale, were recruited. Participants first received either 100 mg/day of milnacipran (33 participants) or 20 mg/day of paroxetine (30 participants), and were then assessed with HAM-D and clinical global impression scale (CGI) for severity of the illness and global improvement, at the beginning and the end of the first, second, fourth, and eighth weeks of the drug treatment. Thirty-eight patients with major depressive disorder completed the study. No statistically significant differences were observed between the two groups in the reduction of HAM-D and CGI scores. However, side effects such as headache and tremor in the first week, psychomotor retardation and difficulty in concentration in the fourth week, and psychomotor retardation in the eighth week of treatment were significantly lower in the milnacipran group, as compared to that of the paroxetine group. We concluded that milnacipran and paroxetine had similar clinical effectiveness during the eight-week treatment of major depressive disorder. Further investigation is needed to examine the clinical suitability of this drug for patients with liver impairments and for elderly patients suffering from major depression. PMID- 19280884 TI - Differential effects of lesions in the subareas of medial prefrontal cortex on the development of behavioral sensitization to amphetamine: the role of environmental context. AB - Based on the notion of the heterogeneity of the medial prefrontal cortex (mPFC), the effects of lesions of the dorsal and ventral subareas of mPFC were examined on the development of behavioral sensitization induced by d-amphetamine (AMP). The first part of this study determined behavioral sensitization to AMP by the use of the rat with intermittent repeated AMP administrations in three types of environmental context including an infrared locomotor activity test-box, the home cage, and a novel third place. In experimental groups, each subject was initially injected with AMP of 0.5 mg/kg as the pretest treatment and then received seven injections of AMP (1 mg/kg) every other day in the context where it was assigned. Following two days of withdrawal, the subject was challenged by AMP of 0.5 mg/kg as the post-test treatment. Behavioral sensitization to AMP on the locomotor activity was determined by the difference between pre- and post-test. The results showed that the most profound locomotor sensitization to AMP appeared in the test box group. A less but significant degree of locomotor sensitization to AMP was observed for the home cage group. A trend of locomotor sensitization to AMP observed in the novel third place group was not statistically confirmed. In the second part of experiment, we then investigated the effects of lesions in the dorsal and ventral mPFC subareas on the development of locomotor sensitization to AMP in the test box and home cage. Results showed that locomotor sensitization was significantly appeared in every sham-operated control group tested in either test box or home cage. Lesions of ventral mPFC significantly inhibited the development of locomotor sensitization to AMP in test box, but not in home cage. Lesions of dorsal mPFC failed to affect AMP locomotor sensitization developed in either test box or home cage. These data indicate that the heterogeneous functions of mPFC subareas involved in the development of behavioral sensitization to AMP are dependent on different contexts applied for repeated intermittent drug administration. PMID- 19280885 TI - Aging effects on glycemic control and inflammation for politicians in Taiwan. AB - Insulin sensitivity is deteriorating with age leading to many metabolic complications, yet fasting glucose is the common metabolic predictor in preventive medicine. In this study we compared the differences in fasting glucose, glucose tolerance, and inflammatory markers between two generations in politically active families. Their physical activity levels and dietary intake amounts were also evaluated. Eight elected councilors and their first order descendents participated in this study. Oral glucose tolerance test (OGTT), insulin, triglyceride, cholesterol, and inflammatory markers including C-reactive protein (CRP) and interleukin-6 (IL-6) were determined. Fasting glucose concentration in politicians was smaller than 100 mg/dL (considered clinically normal), and only approximately 14% concentration difference was observed between two generations. However, all politicians were substantially insulin resistant, compared with their young descendents, evidenced by exaggerated glucose and insulin responses (>100% greater area under curves above baseline) under oral glucose challenged condition. Their waist circumference, diastolic blood pressure, and cholesterol levels were significantly greater than controls. Furthermore, CRP of the politicians was approximately 2.3 folds of the control value suggesting a low grade inflammation. The levels of physical activity and dietary intake were not different between groups. However, the weekly walking energy expenditure for the politician group was approximately 3 times greater than that of the control. CONCLUSION: To reflect the age-dependent metabolic deterioration for the purpose of prevention, OGTT and CRP are far more sensitive measures than fasting glucose value. Greater walking activity in politicians was not sufficient to counterbalance the age-dependent changes. PMID- 19280886 TI - Effects of alcohol on the mouse-killing behavior of olfactory bulbectomized rats. AB - The purpose of the current study was to evaluate the effects of chronic administration of alcohol on the olfactory bulbectomy (OBX)-induced mouse-killing behavior (MKB), an animal model for screening antidepressants. The rats were divided into three groups, which were given alcohol (0, 0.5, or 1 g/kg/day) orally for 28 days. MKB was analyzed before and at the end of each week of the alcohol treatment. The results showed that chronic alcohol treatment produced a significant increase in the latency of MKB, implying that alcohol may have an antidepressant-like activity. This suggests that alcohol dependence or abuse in depressed patients may result from "self-medication". Since it has been reported that OBX causes a decrease in the density of N-methyl-D-aspartate (NMDA) receptors in the brain and that alcohol is a potent and selective inhibitor of NMDA receptors, the possible role of NMDA receptors in this effect is discussed. PMID- 19280887 TI - Acute respiratory distress syndrome associated with hypercalcemia without parathyroid disorders. AB - Acute lung injury (ALI) can be induced by various causes. The occurrence of ALI associated with hypercalcemia has rarely been reported and the mechanisms are unknown. In the present study, we reported the clinical manifestation and pathological findings in patients with hypercalcemia and metastatic calcification. In addition, we addressed the possible mechanism and the preventive strategy for the acute episode of ALI due to hypercalcemic crisis. We encountered five patients with long-term malignancy of various origins. They displayed hypercalcemia and metastatic calcification in the kidney and stomach. One case with transitional cell carcinoma of the urinary bladder developed acute episode of acute respiratory distress syndrome (ARDS). The plasma calcium was elevated to above 5 mM. Simultaneously, He manifested ARDS followed by ALI. The pathological examination revealed severe alveolar edema with multiple calcification. In the other three cases, the plasma calcium level ranged from 3.1 to 4.4 mM and ARDS or ALI did not occur. One patient with esophageal squamous cell carcinoma experienced an acute hypercalcemia (plasma calcium 4.8-5.1 mM) accompanied by ARDS. Corticosteroid and calcitonin were prescribed to reduce the plasma calcium. The symptoms of ARDS also subsided and ALI did not occur. Chronic hypercalcemia results in severe metastatic calcification. The kidney and stomach are the most vulnerable organs. An increase in plasma calcium above 5 mM is a risk factor for developing ARDS and ALI. Our recent experiment in conscious rats and isolated rat's lungs supported this contention. In addition, corticosteroid and calcitonin were able to reduce the plasma calcium and to prevent the occurrence of ARDS and ALI. PMID- 19280888 TI - [Patterns of tooth agenesis. A numerical code for describing dental agenesis]. AB - A genetic or developmental disorder can be the basis for the failure of a normal dental development, resulting in the absence of 1 or more teeth. For genetic research or the development of a plan of treatment, it is important to determine whether certain patterns of dental agenesis are more common than others. In the present article a new method is described where unique values are assigned to certain patterns of tooth agenesis. Using the idea of binary arithmetic, the absence or presence of teeth are represented by 1 and 0, and translated into corresponding unique values, the 'tooth agenesis code' (TAC). This procedure has advantages over existing methods in that it allows for easier data analysis, can contribute to genetic research into the aetiology of tooth agenesis and enables researchers to communicate unequivocally the phenotypes of their studied case. It can also be used in other areas of oral pathology that require pattern recognition. PMID- 19280889 TI - [Prevalence of carotid artery calcification on panoramic radiographs]. AB - In a retrospective study, the prevalence of carotid artery calcifications on routine screening panoramic radiographs of a Dutch Oral & Maxillofacial Surgery department was investigated. Individuals older than 45 without a history of CVA, TIA or head and neck radiotherapy were included.Technically useful radiographs were available for 145 males (mean age 59.6 years) and 303 females (mean age 58.5 years). Both a dentist and an oral maxillofacial surgeon examined these radiographs for radio-opaque lesions adjacent or below the C3-C4 intervertebral space. Forty-two individuals with asymptomatic carotid artery calcifications were identified (9.4% of the study population; mean age 68.2 years): 18 men (12.4%; mean age 66.2 years) and 24 women (7.9%; mean age 69.7 years). In 90% of cases the calcifications were bilateral. These results indicate that panoramic radiographs obtained during the course of routine dental treatment may identify calcified carotid atheromas in asymptomatic subjects. PMID- 19280890 TI - [Treatment of the obstructive sleep apnea syndrome. Side effects of a mandibular advancement device]. AB - The obstructive sleep apnea syndrome is characterised by repeatedly occurring complete or partial obstructions of the upper airway during sleep, which can be accompanied by serious oxygen desaturations. This can result in cardiovascular co morbidity and excessive daytime sleepiness, with an increased chance of motor vehicle accidents and diminished performance at work. The use of a mandibular advancement device appears to be an effective therapy. In the long term, however, the possibility of dental side effects should be taken into consideration. Development of a relative mesio-occlusion has frequently been observed. Side effects are usually mild and transient. To objectively evaluate whether the side effects are stable or progressive, a thorough follow-up is needed. It is therefore desirable that treatments with a mandibular advancement device are carried out by dentists or specialists with experience and special expertise in this area. PMID- 19280891 TI - [Dementia and oral health]. AB - The first part of this article is a translation of an editorial which appeared in the journal Gerodontology. The author warns that a great increase is expected in the number of dementia patients in the United Kingdom and he argues that care for these patients be given a high place on the national agenda. Dementia was also a major issue at the meeting of the International Association for Dental Research in March 2007. Several international studies presented there indicated that elderly people with dementia constitute a group at risk with respect to oral health. In the evaluation of the editorial, the situation in The Netherlands is described. There is also serious concern in The Netherlands about the statistics with respect to dementia. Due to the growing number of frail elderly people having a natural dentition, the need for professional oral care will increase. General practitioners have the important task of providing adequate oral health care for elderly people suffering from dementia who are still living at home. Guidelines for Oral Care, having to do with the improvement of oral care in institutions, appeared recently. With the guidelines, a good basis for developing adequate oral health care of frail elderly people is available. However, the implementation of these guidelines will require some attention. PMID- 19280892 TI - [Delayed wound healing post molar extraction]. AB - One month post extraction of the second left maxillary molar the alveolar extraction site showed no signs of healing and was painful. The patient had been using an oral bisphosphonate during 3 years. Therefore, the lesion was diagnosed as bisphosphonate-induced maxillary osteonecrosis. Treatment was conservative. Since one month later the pain had increased and the wound healing had decreased, a biopsy was carried out. Histopathologic examination revealed a non-Hodgkin lymphoma. PMID- 19280893 TI - [Carpal tunnel syndrome: diagnosis, treatment, prevention and its relevance to dentistry]. AB - Nocturnal pins and needles and other sensory disturbances in the median nerve innervated fingers are caused by local pressure on this nerve in the carpal tunnel. Carpal tunnel syndrome is the most frequently encountered peripheral nerve entrapment. In The Netherlands, the prevalence of carpal tunnel syndrome is estimated 9% among adult women and 0.6% among adult men. Several risk factors have been identified. For dental professionals, the most relevant seem forceful use of the hand during scaling and extractions, use of vibrating ultrasonic equipment and frequent working with the wrist in flexion or in extension. The diagnosis of carpal tunnel syndrome is based on the characteristic complaints, confirmed preferably by abnormal electrophysiological tests. Depending on the degree of impact on daily functioning, treatment for carpal tunnel syndrome may be expectative, conservative or surgical. Adjustment of the working conditions may prevent the development of a carpal tunnel syndrome. PMID- 19280894 TI - The role of bulking agent in pile methane and carbon dioxide concentration during wastewater sludge windrow composting. AB - Wastewater sludge and wood chips were used as feedstock for the construction of two piles, Pile I ("PI") and Pile II ("PII"), at a ratio of 1:1 and 1:2 v/v, respectively. Each pile was originally 1.3-m high, 2.0-m wide, and approximately 9.0-m long. A mechanical turner was used to turn the two windrows every 1 to 2 weeks. Three 500-mL-volume glass funnels were inverted and introduced into each pile: one in the core (named, respectively, "PIC" and "PIIC"), one at the top ("PIT" and "PIIT"), and one at the side ("PIS" and "PIIS"). Every 2 to 3 days, gas samples were collected using gas-tight syringes and analyzed in a gas chromatograph determining carbon dioxide (CO2) and methane (CH4) concentrations. An average gas concentration value between turnings was calculated and a two-way analysis of variance test was used to determine the significance of the differences between piles and pile location, followed by a Post Hoc Tukey test. During the thermophilic period, the mean CO2 concentration in PIC was 103 mL/L, 65 mL/L in PIT, and 24 mL/L in PIS, whereas, for PII, these values were 102mL/L, 59 mL/L, and 24 mL/L, respectively. The mean CH4 concentration between turnings in PIC was 9.2 mL/L, 1.9 mL/L in PIT, and 0.9 mL/L in PIS, whereas, for PII, the corresponding values were 6.4 mL/L, 0.4 mL/L, and 0.1 mL/L. For methane, there were no significant differences between these mean values, not only between the same placement in different piles, but also between different placements and different piles. This is probably due to the relatively frequent turnings (10 turnings during a period of 100 days), which did not allow the development of more anaerobic pockets in PI than in PII, indicating that both piles had similar greenhouse gas impacts. Results for carbon dioxide were similar in both piles, with some differentiation appearing between the core and top placements compared to the side placement. Reduction of the decomposition rate further from the core and a typical windrow chimney effect (gases from the core flowing through the top) explain this similarity between placements. The similarity between piles can be explained by the similar amounts of easily decomposable organic matter found in both piles, indicating that the effect of the bulking agent ratio on the concentration of gases within the pile was not significant. PMID- 19280895 TI - Improved bioconversion of volatile fatty acids from waste activated sludge by pretreatment. AB - Batch tests were conducted to analyze the influence of various pretreatment methods including thermo-alkaline, thermo-acid, ultrasonic-alkaline, and ultrasonic-acid on the bioconversion of volatile fatty acids (VFAs) from waste activated sludge. Experimental results showed that total VFAs (TVFAs) increased by 68.2% for ultrasonic-alkaline and 59.1% for thermo-alkaline. The TVFAs bioconversion in the case of the thermo-acid or ultrasonic-acid pretreatment, however, was lower than that without pretreatment. The results of VFA distribution showed that acetic acid was the most prevalent product, with a fraction of 35.5 to 57.2% for all cases. The mechanism of VFA production was then investigated. Results revealed that, in the two alkaline-pretreated slurries, soluble COD and soluble BOD/soluble COD significantly increased to yield more soluble substrate for the subsequent fermentation. Soluble proteins comprised the main substance provided that was converted to VFAs. A further investigation indicated that volatile (organic) solids, including insoluble proteins, in the solid phase of the samples pretreated with these two methods were hardly consumed during the fermentation. PMID- 19280896 TI - Partitioning and fate of particle-associated E. coli in river waters. AB - Attachment of fecal bacteria to suspended matter in the water column has important implications for its fate in rivers. We examined the part of Escherichia coli (E. coli) associated with suspended matter in natural river water samples, using a combination of 5-microm filtration and beta-D glucuronidase (GLUase) assay to estimate the E. coli abundance and attachment. We observed that the fraction of particle-associated E. coli was positively correlated with suspended matter concentration. The settling rate of particle associated E. coli was found to be positively correlated with suspended matter concentration for samples with suspended matter content lower than 50 mg/L. For samples with higher suspended matter concentration, the settling rate was quite constant (0.066 m/h, on average). In batch experiments using river waters, we observed that free E. coli had a decay rate approximately 2 times higher than particle-associated E. coli. This information can be used to improve the models on the fate of E. coli in rivers. PMID- 19280897 TI - Kinetics of carbaryl degradation by anodic Fenton treatment in a humic-acid amended artificial soil slurry. AB - A Fenton-based indirect electrochemical method, anodic Fenton treatment (AFT), developed for destroying and detoxifying pesticides in the aqueous environment, was evaluated for the degradation of a widely used pesticide, carbaryl, in an artificial soil slurry. More than 90% of carbaryl was removed in less than 20 minutes under given experimental conditions. The effect of initial slurry pH, humic acid content, initial carbaryl concentration, Fenton reagent delivery ratio, and soil/water ratio (w/v) were investigated. The results indicate that humic acid content is the key factor that slows down pesticide degradation, most probably because of its pH buffering and adsorption capacity. A kinetic model, which was shown to fit the experimental data quite well (R2 > 0.99), was developed to describe the carbaryl degradation in the soil slurry during the AFT process. In the presence of humic acid, carbaryl degradation kinetics was found to shift to a pseudo-first-order reaction after an "initiation" stage. PMID- 19280898 TI - Temporal and spatial variations of copper, cadmium, lead, and zinc in Ten Mile Creek in South Florida, USA. AB - Lead (Pb), zinc (Zn), copper (Cu), and cadmium (Cd) often seriously deteriorate water quality. Spatial and temporal fluctuations of the metal concentrations in the Ten Mile Creek (Florida) (TMC) were monitored on a weekly basis at 7 sampling sites, from June 2005 to September 2007. River sediment samples were also collected from these sites in April, June, and October 2006 and January 2007, and analyzed for water, Mehlich 1 (M1), and Mehlich 3 (M3)-extractable metals (Mehlich, 1953, 1984), to examine the role of sediments as sources or sinks of the metals. The concentrations of lead, zinc, copper, and cadmium in the water samples were triethylamine > tributylamine. The intermediates and reaction products were identified by a purge-and-trap gas chromatography mass spectrometer system. The photosensitized dechlorination method degraded PCE via an electron transfer-relay mechanism. Degradation products identified were trichloroethylene, dichloroethylenes, and chloroethylene. It seems a sequential dechlorination pathway was followed. The PCE dechlorination in a natural sunlight irradiation test was shown to be more effective than any of the simulated visible light sources. The result supports the feasibility of future development of solar powered dechlorination remediation systems with the use of sunlight, nontoxic dyes, and electron donors. PMID- 19280903 TI - Occurrence of ciprofloxacin-, trimethoprim-sulfamethoxazole-, and vancomycin resistant bacteria in a municipal wastewater treatment plant. AB - The occurrence of antibiotic-resistant bacteria was evaluated in aqueous samples obtained from a municipal wastewater treatment plant. Samples collected from the influent, clarifier effluent, and disinfected effluent were assayed for fecal coliforms, E. coli, and enterococci exhibiting resistance to ciprofloxacin, trimethoprim-sulfamethoxazole, and vancomycin. Membrane filtration of samples was followed by plating on growth media containing various concentrations of antibiotic. Bacterial colonies on plates with antibiotic exposures greater than the clinical minimum inhibitory concentrations were counted and considered resistant. The numbers of drug-resistant organisms in influent ranged from nondetectable to 7 x 10(5) colony-forming units (CFU)/100 mL for fecal coliforms, nondetectable to 5 x 10(4) CFU/100 mL for E. coli, and nondetectable to 6 x 10(5) CFU/100 mL for enterococci. Fecal coliforms, E. coli, and enterococci with reduced susceptibility to antibiotics were also detected in influent and clarifier effluent; however, the disinfected effluent did not contain resistant bacteria. Species-level identification of enterococci revealed that resistant enterococci were predominantly E. faecalis. PMID- 19280904 TI - Kinetics of p-nitrophenol adsorption by layered double oxides during its hydration. AB - Kinetics of p-nitrophenol (PNP) adsorption onto layered double oxides (LDO) during its hydration into layered double hydroxides (LDH) was studied. Results indicate that the adsorption of PNP onto LDO is a spontaneous endothermic process. The adsorption isotherms correlate well with the Freundlich type model. Results suggest that the adsorption of PNP onto LDO is an entropy-increasing process, and it appears to be in agreement with pseudo-second-order kinetics. Intra-particle diffusion was found to take part in the adsorption processes, and it might be the primary rate-limiting step for the sorbing capacity of LDO to PNP. Results from X-ray diffraction and Fourier transform infrared indicate that PNP molecules are probably taken into the interlayer of the structure during the hydration of LDO into the LDH. Activated carbon was used as a benchmark material in evaluating the sorbing capacity of LDO to PNP. The sorbing capacity of LDO to PNP (32 mg PNP/g LDO) was well below activated carbon (659 mg PNP/g carbon); however, the sorbing process of LDO to PNP is also a unique synthetic process for LDH, which has been used in removing specific anionic species, such as bioagents and pharmaceutical intermediates from waters. PMID- 19280905 TI - How about some teeth for the State Board? PMID- 19280906 TI - Water fluoride concentrations in and around the Greater Houston metropolitan area. PMID- 19280907 TI - Shedding new light on early caries detection. PMID- 19280908 TI - A string of pearls. AB - Most complex interdisciplinary rehabilitative dental cases are associated with tooth eruption due to excessive wear or missing occlusal contacts. As the teeth erupt, the osseous tissues and dentogingival complex extrude with the tooth. Historically, prosthetic treatment demanded placing crowns on short teeth, functional crown lengthening onto root surfaces, or orthognathic surgery to impact teeth. This article will present a more current approach to the treatment of dentoalveolar extrusion. PMID- 19280909 TI - Case of the month. Inflammatory fibrous hyperplasia. PMID- 19280910 TI - Automated external defibrillators in dental offices. PMID- 19280911 TI - [Study on clinico-pathological features of active pulmonary tuberculosis found at autopsy in a general hospital]. AB - OBJECTIVES: To clarify clinico-pathological features of tuberculosis found at autopsy. METHODS: This study investigates 18 (3.7%) of active pulmonary tuberculosis out of 489 autopsy in Tachikawa Sougo Hospital during the period from 1992 to 2005. RESULTS: There were 11 men and 7 women, with a median age of 69.5 years. Tubercle bacilli were proved from sputum in 6, which consisted of 3 with positivity on sputum smear microscopy and culture, and 3 with positivity only on sputum culture. Two were examined, but not diagnosed before death. Three didn't show any positive result despite of repeated sputum tests. The features of the chest radiological findings were: (1) Shadows that present prior tuberculosis (ex: nodules, fibrotic lesion) were found in 9 and ground-glass-opacity in 5. (2) In 6, radiological findings consistent with tuberculosis were not pointed out because shadows such as fibrosis, pleural effusion, or cancer were mixed in the same lung. (3) In 11, main radiological findings were found in atypical segments, when there were some underlying conditions such as the use of corticosteroidal therapy or diabetes mellitus. Four were diagnosed correctly, and treated with anti-tuberculosis drugs. Other 14 were not diagnosed before death and diagnosed wrongly as pneumonia, cancer, or other diseases. Encapsulated caseous nodules were seen in 7, and autopsy confirmed that 12 including these 7 were caused by endogenous reactivation. Miliary tuberculosis was found in 5, caseous pneumonia/bronchitis in 6. One had tuberculous empyema. As to underlying diseases, 8 had malignant disease, 6 had diabetes mellitus and 6 were treated with corticosteroids. CONCLUSION: This study suggests that sputum culture or radiological findings are not sufficient enough to diagnose tuberculosis, especially in compromised host. We emphasize the vital role of treatment for latent tuberculosis for cases with high risk of endogenous reactivation, and it's necessary to make the guideline for the treatment of such latent tuberculosis. PMID- 19280912 TI - [Situation analysis of the monitoring of tuberculosis infection among hospital staff]. AB - PURPOSE: To investigate monitoring of new tuberculosis infection among staff in hospitals running the model-beds for tuberculosis patient care. OBJECT & METHOD: Questionnaires were sent to the hospitals and the responses were analyzed. RESULT: Valid answer sheets were recovered in 51% (38/75). Forty percent of hospitals performed periodic monitoring of new tuberculosis infection among staff. Fifty percent performed only one test for tuberculosis infection on employment but did no perform further periodic tests. Ten percent did not perform specific tests for tuberculosis infection at all. There were very few persons who were prescribed preventive therapy based on test results for tuberculosis infection. CONCLUSION: Less than half of the hospitals performed periodic monitoring of new tuberculosis infection, but there were few tuberculosis cases among hospital staff. It is necessary to investigate further the efficacy and necessity of periodic monitoring of new tuberculosis infection among hospital staff. PMID- 19280913 TI - [Findings and ongoing research in the molecular epidemiology of tuberculosis]. AB - SETTINGS: While in The Netherlands the organization of tuberculosis (TB) control in the era of elimination is being discussed, worldwide TB is still a major problem. OBJECTIVES: To highlight findings and ongoing research in the molecular epidemiology of tuberculosis. FINDINGS: In the past decade, many studies have pointed out the significance of the Beijing strains in the worldwide tuberculosis epidemic. In South-East Asia, as well as the former USSR Republics, the Baltic States and South Africa, Beijing strains are emerging and associated with (multi drug) resistance. Recent research in China has pointed out that the average proportion of Beijing genotype strains in 10 provinces amounts 73%, but varies significantly by region. The highest density was observed in the Beijing region; 93%. Currently, research is focused on the evolutionary development of Beijing strains. Two evolutionary lineages were distinguished and the "modern" lineage seems associated with spread, despite BCG vaccination in Vietnam. A project of the European Union with 11 international partners is trying to disclose the evolutionary adaptation of M. tuberculosis in response to vaccination and exposure to anti-tuberculosis drugs. Also in Europe the interest in the Beijing genotype has recently been strongly awakened. While 6-7% of the susceptible isolates are of this genotype, 85% of the MDR-TB transmission in Europe can be attributed to the spread of Beijing strains. Moreover, one cluster of 174 MDR-TB cases has been detected and this re-emphasizes that particular genotypes are capable of gaining poly-resistance without losing fitness. It seems, therefore, important to have more studies on the dynamics in the population structure of M. tuberculosis worldwide. In the presentation examples of such studies will be discussed. Since 1993, IS6110 restriction fragment length polymorphism typing has gained recognition as the gold standard in the molecular epidemiology of tuberculosis. However, although the application of this method has brought significant new insights into the transmission of tuberculosis, it remains technically demanding and time-consuming. Therefore, recently a new standard typing method, the variable numbers of tandem repeat (VNTR) typing, has been proposed in a publication in the J. Clin. Microbiol. In the Netherlands, a large part of the collection of nearly 15,000 M. tuberculosis isolates are currently being retyped with VNTR typing. This is not only meant to evaluate the utility of the new 24 loci VNTR typing, but also to conserve the epidemiological trace of tuberculosis transmission in the past. The first results will be discussed. The role of non-tuberculous mycobacteria (NTM) in the diagnosis and treatment of tuberculosis-like diseases becomes increasingly important in the era of the HIV/AIDS pandemic. In Zambia it was recently shown that only in 50% of the cases diagnosed by Ziehl-Neelsen microscopy and X-ray, M. tuberculosis was isolated in a liquid culture system. In the other cases often only NTM can be cultured. In The Netherlands, particular NTM like M. avium, frequently cause lymphadenitis in children. This infection is normally indicated as "bird tuberculosis". However, with IS1245 RFLP typing it was pointed out that the M. avium isolates from lymphadenitis cases do not represent the bird type M. avium strains, but the M. avium strains typically found in humans and slaughtered pigs: Mycobacterium avium hominissuis. PMID- 19280914 TI - [The cause of autoimmunity]. PMID- 19280915 TI - [Historical review of rheumatology and future directions]. AB - Rheumatic diseases are suggested to be recognized 2400 years ago, but little progress had been made in distinguishing specific diseases until the 17th century. After the two decades, the concept of rheumatic diseases was established and followed by the new concept of autoimmune diseases in the 20th century. Many key drugs for rheumatic diseases, such as glucocorticoids and immunosuppressants, were developed in the 1940s and 1950s. Their effect, toxicity and limitations have been investigated for fifty years. On the other hand, new treatment strategy was developed. Advances in genetics and molecular cell biology in the 1980s brought us the discovery of many cytokines and chemokines and elucidated the role of these molecules in autoimmune diseases. These progresses have led to the development of a number of biological agents for the rheumatic diseases during past decade. Among these drugs, TNF blockade showed the remarkable effectiveness in rheumatoid arthritis and this success indicated the new treatment era. However, compared to the simplicity of target molecules in monoclonal antibody therapy, biological effects are more complex. Therefore, we must keep in mind the possibility of the appearance of unexpected adverse effects such as TGN1412. This review presents a history of rheumatology and discusses future directions of basic and clinical research for rheumatic diseases. PMID- 19280916 TI - [Epidemiology of collagen diseases]. AB - Collagen diseases have been generally thought to be rare, but the number of patients with rheumatoid arthritis(RA) becomes over 700,000 in recent days. In addition, the survival rate has markedly increased, but many lupus patients are still suffering from so called intractable organ involvements such as severe lupus nephritis and CNS lupus, etc. The main purposes of the treatment for RA are to prevent bone/cartilage destruction, to improve the quality of life(QOL) of patients, and to improve prognosis. For these purposes, new type of drugs such as biologic agents are introduced, and showing remarkable effects comparing with conventional DMARDs. But the side effects and cost of these agents are becoming problems. To overcome these problems, the epidemiology that can clearly show the benefits of these agents on daily life of patients is becoming much more important issue. PMID- 19280917 TI - [Pathogenesis of autoimmunity]. PMID- 19280918 TI - [Genome-wide association studies: recent advances and future directions]. AB - Information about human genetic variation collected by the HapMap project and recent advances in high-throughput genotyping have made genome-wide association studies (GWAS) technically feasible. In the past two years, through GWAS using commercially available SNP-typing platforms, numerous loci contributing to disease risk have been identified for common diseases including rheumatoid arthritis and systemic lupus erythematosus. From such successful studies, human geneticists have gained not only broad consensuses in methodological and technical issues, but also clear recognition about the current limitations in the common SNP-based GWAS strategy. Large-scale projects are ongoing to comprehensively catalogue genome structural variants and rare SNPs in multiple human populations. Such a deeper catalogue of genetic variation will accelerate the identification of genetic risk variants associated with common diseases. PMID- 19280919 TI - [B lymphocyte abnormalities in connective tissue disorders]. AB - In addition to the precursors of Ab-secreting cells, B lymphocytes have more essential functions in regulating immune responses than had previously been appreciated. B cell functions include antigen-presenting cells, production of various cytokines, and differentiation of T effector cells. Abnormalities of these B cell functions could contribute to the induction or development of autoimmunity in systemic autoimmune disorders. In this regard, the importance of B cells in systemic autoimmunity and disease expression has been emphasized; therefore, various B cell targeting therapies will be applied to connective tissue disorders more intensively. PMID- 19280920 TI - [Contribution of Th-1, Th-2, Th-17 or regulatory T cells to connective tissue diseases]. AB - Connective tissue diseases (or systemic autoimmune diseases), such as rheumatoid arthritis or systemic lupus erythematosus, are characterized by B cell hyperactivity and production of various autoantibodies. T cells help B cell activation in the germinal center by enhancing somatic hypermutation and generation of high affinity pathogenic autoantibodies. Previously helper T cells were divided into Th-1 and Th-2 cells. Recently, Th-17 cells and regulatory T cells have been identified as distinct T cell lineages and their roles in inflammation or immune regulation are under intensive investigation. In this review, we discuss the contribution of each T cell subset to autoantibody production and systemic autoimmune diseases. PMID- 19280921 TI - [Role of TLR-dependent and independent pathways in autoimmunity]. AB - Innate immune system senses invading pathogens through their molecular patterns and elicits pleiotropic immune responses including production of proinflammatory cytokines and antigen presentation. However, the system could sense host endogenous molecules and elicits immune response against self resulting into autoimmunity. In this regard, Toll-like receptor (TLR) is involved in several autoimmune diseases such as systemic lupus erythematosus. Recently identified TLR independent pathways are revealed also to have roles in autoimmunity as well as TLR. Innate recognition of intracellular DNA, of which receptor is still unknown, is involved in several autoimmune diseases through its endogenous ligand. Moreover, Nod-like receptors play important role in chronic inflammatory diseases like gout and Crohn's disease. In this review, the authors described the role of innate recognition system in autoimmunity, emphasizing on their molecular mechanism. PMID- 19280922 TI - [Rheumatoid arthritis]. AB - Pathogenesis of rheumatoid arthritis is likely to implicate anti-citrullinated protein/ peptide antibody(ACPA) and an immunodistortion including abnormal T cell subpopulation. Based on above and other recent findings, new biological agents targeted to inflammatory cytokines such as tocilizumab, activated T cells (abatacept) or B cells (ocrelizumab), as well as new small molecule drugs such as JAK3 inhibitor, are sure to further facilitate remission without impaired activity of daily life in patients with RA. The contribution of Japanese physician-scientists to the progress in rheumatology has been significant as described in this review, and it must be increasingly greater in the near future. PMID- 19280923 TI - [Recent investigations on the basis of pathogenesis of SLE and new therapeutic approaches]. AB - Systemic lupus erythematosus (SLE) is a multisystem, autoimmune, connective tissue disorder with a broad range of clinical presentations. SLE predominantly affects women, especially from ethnic groups with African and Asian ancestry. This disorder is a chronic illness that can be life threatening when major organs are affected, but more commonly results in debilitating condition and affects employment and fertility. This article summarizes the recent advances in our understanding of the genetics, epidemiology and pathogenesis of SLE. In addition, progress in the assessment and management of serious complications such as lupus nephritis and CNS lupus is reviewed. New therapeutic approaches, such as low-dose cyclophosphamide regimens, mycophenolate mofetil and biological agents are also discussed. PMID- 19280924 TI - [Vasculitis syndrome]. AB - The recent development of biologic therapies capable of selectively targeting components of the immune system has revolutionised the treatment of inflammatory arthritides. The increase in use of biologic agents coupled with the expansion in the knowledge of the pathogenesis of vascular inflammation has led to their application in the treatment of primary systemic vasculitis. Biological therapies appear to have a place in the therapeutic strategy for ANCA-associated systemic vasculitides, at least for patients whose disease is refractory to conventional therapy. The use of biologics as targeted therapies has also, in reverse, improved our understanding of the pathophysiology of vascular inflammation. However, the precise indications for TNFalpha inhibitors or anti-CD20 monoclonal antibodies have not yet been defined. These biologics must be prescribed extremely cautiously and only in trial settings, especially in view of the adverse effects. PMID- 19280925 TI - [Sjogren's syndrome]. AB - Sjogren's syndrome (SS) is chronic autoimmune disease characterized by destructive lymphocyte infiltration of salivary and lacrimal glands, which results in dry eyes and dry mouth. Despite extensive study of the underlying cause of SS, the pathogenesis remains obscure. The Sjogren's International Collaborative Clinical Alliance (SICCA) by five Research Groups located in Argentina, China, Denmark, the United States and Japan, is the first registry and clinical data-specimen repository to establish the International Standard Criteria for diagnosing SS. Patients with SS have a relative increased risk for development of B cell lymphoma compared with other autoimmune rheumatic diseases. We discuss the possible mechanisms for lymphoma development. The topics concerning SS is IgG4-related lymphoproliferative diseases, such as Mikulicz's disease and autoimmune pancreatitis. Based on the analysis of patients registered from all over Japan, we propose a new clinical entity IgG4-positive multi-organ lymphoproliferative syndrome (IgG4+ MOLPS). PMID- 19280926 TI - [Systemic sclerosis]. AB - Systemic sclerosis (SSc), the focus of this review, is a generalized connective tissue disease that involves sclerotic changes in the skin and sometimes various other organ systems. Clinical outcomes have improved probably due to better management of the complications, but SSc is still considered to be incurable and diffuse cutaneous SSc carries high risk of fatality. Although the pathogenesis of SSc is still unknown, the basic mechanism appears to involve endothelial cell injury, overproduction of extracellular matrix (ECM), and aberrant immune activation. This review discusses recent studies that investigated the cellular and molecular mechanisms in the pathogenesis of SSc. PMID- 19280927 TI - [Recent advance in polymyositis and dermatomyositis research]. AB - Past paradigm that the muscle tissue injury is driven by CD4 T cells and associated humoral immunity in dermatomyositis (DM), and by cytotoxic T cells in polymyositis (PM) is now under challenge. Although pathogenic autoantigens are to be identified, skeletal muscle C-protein was an excellent immunogen to provoke experimental myositis mimicking human PM. Serum antibodies against aminoacyl tRNA synthases appear in PM/DM patients, but more often in interstitial pneumonitis patients. Inflammatory cytokines are obviously involved in the pathogenesis. Animal studies showed that autoimmune myositis occurs without tumor necrosis factor alpha. Indeed, its blockade has yielded inconsistent outcome. Most crucial ones will be therapeutic targets in the future. PMID- 19280928 TI - [Neuropsychiatric lupus (CNS lupus and PNS lupus)]. AB - Patients with SLE show in a variety of neuropsychiatric symptoms, although we could not use standardized methods for evaluating and making diagnosis of the syndromes. ACR felt to develop objective and valuable tools for the diagnosis and classification of neuropsychiatric lupus, therefore, they proposed a new tentative set of nomenclatures describing neuropsychiatric lupus syndromes in accordance with 4th edition of Diagnostic and Statistical Manual of Mental Disorders provided by American Psychiatric Association. For this purpose, The Ad Hoc Committee collected and evaluated 108 case presentations of neuropsychiatric lupus from USA, Canada, and UK. Re-evaluation of the tentative nomenclatures selected 19 neuropsychiatric syndrome to facilitate and enhance clinical research. They grouped into peripheral and central nervous system lupus, and central nervous system lupus was divided into neurologic syndromes and diffuse psychiatric/ neuropsychological syndromes. Instead of organic brain syndrome, a term of acute confusional state was introduced. PMID- 19280929 TI - [Pulmonary involvements in patients with collagen disease]. AB - Pulmonary involvement is a common future in patient with collagen diseases. Some of the pulmonary involvements are a resistant of current available treatment and a fetal condition. These conditions include interstitial pneumonitis in scleroderma, acute diffuse alveolar damage (DAD) in polymyositis/dermatomyositis (PM/DM), alveolar hemorrhage, and pulmonary hypertension in collagen disease. Intensive immunosuppressive treatment in early stage of interstitial pneumonitis may be effective for an intractable condition. Oral cyclophosphamide therapy is effective for non specific interstitial pneumonitis of scleroderma in a randomized control trial. Acute interstitial pneumonitis, so called DAD in patients with PM/DM is a resistant of steroid therapy and a poor prognostic complication. Combination therapy with high dose corticosteroids and immunosuppressive drugs (cyclosporin A, cyclophosphamide) is sometimes available therapy. Diffuse alveolar hemorrhage sometimes occurred in collagen disease including antineutrophilic cytoplasmic antibody (ANCA) associated vasculitis, systemic lupus erythematosus. Intensive treatment need to inhibit hemorrhage. PMID- 19280930 TI - [Cardio-pulmonary disorders associated with collagen vascular disease]. AB - Among the cardio-pulmonary disorders associated with collagen vascular disease (CVD), pulmonary arterial hypertension (PAH) is recognized as most serious problem. Because PAH is difficult to manage and its prognosis is terribly poor. Thus, early diagnosis of PAH in CVD is important. Since PAH patients associated with CVD are often asymptomatic in their early stage, screening with echocardiography is needed. The degree of inflammation causing PAH through endothelial injury is closely associated with CVD, and thus, the improvement of PAH may be achieved by immunosuppressant, especially in its early stage. Once the diagnosis of PAH is confirmed, treatment with oral vasodilators should be started as soon as possible. If oral vasodilators prove inadequate, intravenous epoprostenol should be administered without hesitating. PMID- 19280931 TI - [Renal involvement in connective tissue diseases]. AB - Prognosis of the renal involvement of connective tissue diseases such as systemic lupus erythematosus or systemic sclerosis is getting better due to the induction of the new immunosuppressant such as tacrolimus and hypotonicas which reduce glomerular hypertension such as angiotensin converting enzyme inhibitor or angiotensin II receptor antagonist. Since patients with myeloperoxidase-anti neutrophil cytoplasmic antibody related glomerulonephritis are older than those with Wegener's granulomatosis, the strong immunosuppressive treatment recommended in Western countries should be avoided. The treatment guideline issued by the Japanese Society of Nephrology is suitable for elderly Japanese patients. Recently, there have been some reports of the use of mizoribine, which is a mild immunosuppressant drug. PMID- 19280932 TI - [Evidence-based laboratory tests for the diagnosis and evaluation of connective tissue diseases]. AB - For the proper application of the laboratory tests, it is essential to select the best suitable ones based on the inquiry and the physical examination of the patients and to interpret the test results in accordance with the evidence-based manner. Here described is the way to make use of the clinical laboratoy tests, not only the immunological ones such as rheumatoid factor and anti-nuclear antibodies but the general screening tests such as complete blood cell counts, focusing mainly on the diagnoses of the connective tissue diseases. PMID- 19280933 TI - [Diagnostic imaging of systemic rheumatic diseases]. AB - In patients with systemic rheumatic diseases, imaging often plays an important role in diagnosis. Erosions of cartilage and bone are among the cardinal features of RA. Plain radiography is used most often to assess for the presence of joint damage, however, other imaging techniques such as contrast-enhanced magnetic resonance imaging (MRI) and ultrasonography are valuable to detect early inflammatory change of joint. In addition, chest CT is essential to detect pulmonary involvement which sometimes can be a main prognostic factor. Finally, arteriograms are helpful in identifying and characterizing a vasculitis of large and medium-sized arteries, in whom an obvious area for biopsy is absent. The present article reviews these imaging techniques. PMID- 19280934 TI - [How to find a collagen-vascular disease patient in an early stage and when to let her/him visit a rheumatologist]. AB - Collagen-vascular diseases could be defined as 'chronic', 'inflammatory', 'multiorgan diseases' 'with immunological derangement'. It is important to know that these diseases frequently involve locomotorium, skin, and vessels, of which symptoms or signs are easily sensed by patient herself or observed by herself or a doctor from her body surface. Even among routine laboratory findings, cytopenia, lymphocytopenia, inflammatory reactant level increase, serum total protein level increase could be indicative of these diseases. When considering a substantial number of patients with these diseases or with their subclinical status, routine medical check for rheumatoid factor or anti-nuclear antibody would be better recommended. In case that differential diagnosis is difficult or a strong intervention may be necessary, visiting a rheumatologist at least once would be a choice for early diagnosis or to avoid iatrogenic adverse events induced by irrelevant steroid, disease-modifying anti-rheumatic drug or immunosuppressant. Socio-economically, the early diagnosis and early treatment for these diseases would be very effective showing a good cost-performance. PMID- 19280935 TI - [Nonsteroidal anti-inflammatory drugs and glucocorticoids for treatment of connective tissue diseases]. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) and glucocorticoids (GCs) are highly valuable medication available to a physician. The clinical usefulness of NSAIDs and GCs in the management of connective tissue diseases were discussed in this review. We summarized serious adverse reactions of these drugs. In addition, recent informations of these drugs in clinical use are also introduced. PMID- 19280936 TI - [Immunosuppressants]. AB - In recent years, novel immunosuppressive drugs applies to various autoimmune diseases and the evidences increase. Since immunosuppressive drugs inhibit nonselectively the whole immune system, there are possibilities that infection may be induced by decreasing the immunity against pathological agents and malignancy may develop by suppressing immune surveillance. In order to use the immunosuppressants safely and effectively, we should obey the indication and dosage, and be familiar with the side effects. It is also important to take informed consent from patients after explaining the benefit and risk. Therefore, the doctors who are going to use immunosuppressive drugs must know well about the indication, the side effects, the metabolism and the action mechanisms of those drugs. PMID- 19280937 TI - [Targeted therapies including monoclonal antibodies for connective tissue diseases]. AB - Recent advance of targeted therapies including monoclonal antibodies and fusion proteins has allowed effective strategies in the treatment of rheumatoid arthritis. And now, TNF inhibitors are broadly used for rheumatoid arthritis and prevent the disease progression. Meanwhile, B cell targeted therapies and anti interleukin-6 receptor antibody treatment are not only used for second line biological agents for rheumatoid arthritis, but also expected for the treatments of various autoimmune diseases. Recent year, some of novel small molecules, which inhibit the signal transduction of various surface receptors of immune cells, are in clinical trials. These drugs will be a breakthrough for the treatment of some autoimmune disorders. PMID- 19280938 TI - [Biological agents]. AB - There are two types of biological agents for the treatment of rheumatoid arthritis (RA); monoclonal antibodies and recombinant proteins. Among the latter, etanercept, a recombinant fusion protein of soluble TNF receptor and IgG was approved in 2005 in Japan. The post-marketing surveillance of 13,894 RA patients revealed the efficacy and safety profiles of etanercept in the Japanese population, as well as overseas studies. Abatacept, a recombinant fusion protein of CTLA4 and IgG, is another biological agent for RA. Two clinical trials disclosed the efficacy of abatacept for difficult-to-treat patients: the AIM for MTX-resistant cases and the ATTAIN for patients who are resistant to anti-TNF. The ATTEST trial suggested abatacept might have more acceptable safety profile than infliximab. These biologics are also promising for the treatment of RA for not only relieving clinical symptoms and signs but retarding structural damage. PMID- 19280939 TI - [Intravenous immunoglobulin (IVIG)]. AB - Intravenous immunoglobulin (VIG) was given by Imbach in 1981 to idiopathic thrombocytopenic purpura (ITP). The cure of many of the patients in response to the IVIG expanded the use of IVIG to other autoimmune condition. The diseases proven the efficacy of VIG by controlled trial were ITP, Guillain-Barre syndrome, chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), multiple motor neuropathy, multiple sclerosis, myasthenia gravis, ANCA related vasculitis, Kawasaki disease, autoimmune uveitis and dermatomyositis. There are several uncontrolled trials and case reports about the efficacy of IVIG on the connective tissue diseases, such as systemic lupus erythematosus, systemic sclerosis, Sjogren syndrome, antiphospholipid antibody syndrome and Still's disease. In Japan, only ITP, Kawasaki disease and CIDP are allowed to be use IVIG by health insurance. I would like to summarize the application of VIG on collagen diseases and refer both novel mechanisms as well as to complications of this therapy. PMID- 19280940 TI - [Therapeutic apheresis for rheumatic diseases]. AB - The main stream of treatment for rheumatic disease is pharmacotherapy. Corticosteroids, DMARDs, the immunosuppressive agents and biologics are the gold standard of the treatment. However, some cases showing resistance to these treatments exist, and the potential for side effects exists in all of these pharmacotherapies. Furthermore, there are cases in which pharmacotherapy cannot be effectively used due to organ derangement. Apheresis is totally different from pharmacotherapy as a concept, and it is a treatment to control disease activity by removing the factors which contribute to the pathogenicity. The synchronized therapy of apheresis and pharmacotherapy is the treatment that may result in a more beneficial effect of treatment. This review introduces the experiences as an overall approach to better, safer management for rheumatic diseases. PMID- 19280941 TI - [Vascular regeneration therapy for intractable vasculitis]. AB - The field of cell-based transplantation has expanded considerably and is poised to become an established cardiovascular therapy in the near future. In this review, we will focus on endothelial progenitor cells (EPCs), which are immature cells capable of differentiating into mature endothelial cells (ECs). The major source of EPCs is the bone marrow (BM). BM-derived EPCs are mobilized into peripheral blood and recruited to the foci of pathophysiological neovascularization and reendothelialization, thereby contributing to vascular regeneration. Severe EPC dysfunction is an indicator of poor prognosis and severe endothelial dysfunction. Indeed, number of circulating EPCs and their migratory activity are reduced in patients with diabetes, coronary artery disease (CAD) or subjects with multiple coronary risk factors. Effective neovascularization induced by EPC transplantation for hindlimb, myocardial and cerebral ischemia has been demonstrated in many preclinical studies, and early clinical trials of EPC transplantation in chronic and acute CAD as well as Buerger disease indicate safety and feasibility of the cell-based therapies. Further development in cell processing technology for efficient isolation, expansion, mobilization, recruitment and transplantation of EPCs into target tissues are underway and expected to be tested in clinical trials in the near future. PMID- 19280942 TI - [Recent progress in clinical trials for rheumatic diseases]. AB - Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disease that causes significant morbidity and mortality. RA patients should be started with DMARD represented by methotrexate (MTX) as early as possible. However, even use of MTX often fails to control disease activity and to prevent structural damage and, thereby, more effective treatment strategies are needed. Since TNF-alpha and IL-6 play a pivotal role in the pathological processes of RA, biologics targeting these cytokines with MTX, have revolutionized the treatment of RA, producing significant improvement in clinical, radiographic and functional outcomes not seen previously. However, even with these drugs the frequency and degree of responses are restricted. Therefore, new agents targeting cell surface molecules which are involved in cellular interaction and/or signaling on immune cells have been emerging, in order to increase response rates and to achieve high frequencies of remission or even cure. Two biologics abatacept, a CTLA4-Ig fusion protein, and rituximab, an anti-CD20 antibody, were launched in US and EU for the treatment of RA and many biologics are under the clinical trials from the similar concept. Thus, certain biologics have brought about paradigm shift in the treatment of rheumatic diseases, but an economical issue remains unsolved. In order to overcome the concern, low molecular weight chemical products have been rethought. Not a few agents targeting intracellular activation signaling in immune cells such as Jak and Syk are under clinical examinations and some of them appear to show wonderful results, which are comparable to biologics in the context of the treatment of rheumatic diseases. The prospects are here. PMID- 19280943 TI - [Autoinflammatory syndrome]. AB - The autoinflammatory syndromes include a group of inherited diseases that are characterized by 1) seemingly unprovoked episodes of systemic inflammations, 2) absence of high titer of autoantibody or auto-reactive T cell, and 3) inborn error of innate immunity. In this article, we will focus on the clinical features, the pathogenesis related the genetic defects, and the therapeutic strategies in the representative disorders including familial Mediterranean fever (FMF), TNF receptor associated periodic syndrome (TRAPS), cryopyrin-associated periodic syndrome (CAPS), hyper-IgD with periodic fever syndrome (HIDS), syndrome of pyogenic arthritis with pyoderma gangrenosum and acne (PAPA), and Blau syndrome. Recent advances in genetics and molecular biology have proceeded our understanding of the pathogenesis of autoinflammatory syndromes. PMID- 19280944 TI - [Appendicular thermal therapy for heart failure]. AB - We preliminarily assessed the utility of a steam foot bath in 4 male patients with dilated cardiomyopathy with refractory chronic congestive heart failure awaiting heart transplantation. Sublingual temperature significantly increased from 36.1 +/- 0.3 to 36.7 +/- 0.2 degrees C (p = 0.01). Blood pressure, heart rate and numbers of ventricular arrhythmia did not differ before and after two weeks of the therapy. Grade of mitral regurgitation tended to decrease (2.0 +/- 1.2 to 1.3 +/- 1.0, p = 0.06), and endothelial function measured by arterial tonometry significantly increased from 1.35 +/- 0.26 to 2.13 +/- 0.26 (p = 0.02). Thus, steam foot bath therapy may be safe and beneficial for the patients with endstage heart failure awaiting heart transplantation. PMID- 19280945 TI - [Assessment of proximal aortic anastomosis device in coronary artery bypass grafting]. AB - The aim of this study was to assess the feasibility and safety of a new proximal anastomotic device (PAD) "Enclose II" in coronary artery bypass grafting (CABG). PAD enables the construction of a proximal aortic anastomosis without the use of partial clamp of the ascending aorta, thus reduces the incidence of adverse perioperative neurologic injury related to atheroembolic events. This device was used in 41 off-pump CABG and 11 on-pump beating heart CABG patients for performing 46 radial artery (RA) and 9 vein anastomoses to the aorta. The subjects were 43 males and 9 females, with a mean age of 63.6 years. Thirteen (25%) patients had severe atherosclerotic cerebrovascular lesions preoperatively. The mean flow in the RA graft was 52.4 +/- 26.9 ml/min and that of saphenous vein graft (SVG) was 61.1 +/- 31.9 ml/min. Angiography showed all grafts patent. There was no procedure-related adverse events or cerebrovascular complication. Enclose II device can be a valuable tool to perform RA and vein anastomoses in CABG. PMID- 19280946 TI - [Video-assisted thoracoscopic lobectomy for congenital pulmonary airway malformations (type II congenital pulmonary airway malformations) in a 1-year-old girl]. AB - Video-assisted thoracic surgery (VATS) is increasingly being used for congenital cystic lung diseases but there are very few reports of VATS lobectomy because of its technical difficulties. Congenital pulmonary airway malformations (CPAM) is a rare congenital developmental abnormality; an immediate and proper surgical resection must be performed for prevention of malignant transformation and recurrent pulmonary infection. Here, we report a case of a 1-year-old girl with type II-CPAM who was successfully treated with VATS lobectomy. Under single lung ventilation, the child was placed in the lateral decubitus position. A skin lateral incision approximately 3 cm long was made and a 5th intercostal thoracotomy from the mid-axillary line was performed. A 5 mm thoracoscope was introduced through the 7th intercostal space at the midaxillary line. Naruke's forceps were used for the ligation of the major vessels and the basal bronchus was closed with continuous sutures. JMS swabs were effective for the stripping of pulmonary vessels sheaths. An ultrasonic cutting and coagulating surgical device was used to incise the incomplete fissure. Histological examination showed type II-CPAM of the right lower lobe accompanied with lung abscess and S10 hypoplasia. For a certain category of patients, surgical lobectomy using the thoracoscopic approach can be a safe and effective treatment for infants with CPAM. PMID- 19280947 TI - [Clinical analysis of resected pulmonary pleomorphic carcinoma]. AB - OBJECTIVE: We studied the clinical feature of pulmonary pleomorphic carcinoma. PATIENTS: Of 1,073 patients underwent the resection of lung cancer in our hospital, 15 patients (1.4%) had primary pulmonary pleomorphic carcinoma. RESULTS: All patients were male, and the age was ranged from 57 to 83, mean 70.3 years old. The mean Brinkman Index was 1,028.7. Thirteen patients underwent the lobectomy with lymph node dissection, 1 patient underwent the bilobectomy with lymph node dissection, and the wedge resection was done for the remaining one. Four patients were in stage IA, 5 were in stage IB, 2 were in stage IIB, 1 was in stage IIIA, 2 were in stage IIIB, and 1 was in stage IV. Histologically, the vascular invasion was showed in 8 of 15 patients. The recurrence was occurred on 9 patients, from 1 to 20 months after surgery. The site of recurrence was lung in 3 patients, brain in 3, and abdominal lymphnodes, subcutaneous soft tissue of shoulder, carcinomatous pleurisy was observed in one patient each. The 1-year survival was 62.5% , and the 3-year survival was 46.9%. Conclusion : The prognosis for pulmonary pleomorphic carcinoma is poor. More meticulous follow up is necessary for patients with pleomorphic carcinoma than those with common non small cell carcinoma. PMID- 19280948 TI - [Extended septoplasty for left ventricular outflow tract obstruction after the intracardiac repair of double outlet right ventricle]. AB - A 14-year-old boy with a left ventrivular outflow tract obstruction (LVOTO) after the repair of double outlet right ventricle (DORV) at the age of 7 months was successfully treated with an extended septoplasty. A significant pressure gradient occurred between the left ventricle and the ascending aorta of 70 mmHg in spite of the previous surgery for LVOTO with muscle resection and the ventricular septal defect (VSD) patch enlargement at the age of 5. An extended septoplasty was performed including extensive resection of the conus muscle and the previously placed patch, long incision on the interventricular septum toward the apex, and reconstruction of the outflow tract using a Hemashield patch. The cathterization 10 months after the operation revealed an effective relief of LVOTO with left ventricle (LV) and aorta (Ao) pressure of 115 and 103 mmHg, respectively. An extended septoplasy is effective for the relief of LVOTO, but further follow-up is mandatory of both the left ventricular function and mitral valve function. PMID- 19280949 TI - [Multiple recurrent cardiac myxomas]. AB - A 49-year-old male had undergone resection of biatrial multiple myxomas 4 years previously in another hospital. Echocardiography revealed new masses in the left atrium and the right ventricle. Both tumors were removed surgically and subsequently treated with adjuvant chemotherapy. There was no further recurrence during the following 4 years. It was considered that the multiple recurrences in our case were due to multicentricity of the tumor. PMID- 19280950 TI - [Surgically treated pleomorphic carcinoma of the lung]. AB - We experienced 3 resected cases of pleomorphic carcinoma of the lung. Each cases were 74-year-old man (case 1), 74-year-old woman (case 2) and 69-year-old man (case 3). Two patients (case 1 and 2) were histologically diagnosed as pleomorphic carcinoma composed of spindle cell carcinoma with giant cell carcinoma. One patient (case 3) was similarly diagnosed as pleomorphic carcinoma composed of spindle cell carcinoma with adenocarcinoma and squamous cell carcinoma. Although lymph nodes metastasis were not recognized in all patients, invasion to vessels were recognized in 2 patients (case 1 and 3). In one patient (case 1), recurrence was recognized at contralateral side 1 month after surgery and he died of other disease 2 months after surgery. The other 2 patients were alive without recurrence 24 and 5 months after surgery. Recently it is reported that recurrence is recognized at early phase after surgery and prognosis is poor in a case with vessel invasions in spite of pathological NO state. Since one patient (case 3) had nonmetastatic lymph nodes with vessel invasions, careful observation is considered to be necessary. PMID- 19280951 TI - [Gastrointestinal hemorrhage induced by percussive ventilator in an infant with chronic lung disease complicated after surgery for corrected transposition of the great arteries]. AB - A newborn patient (birth weight 2,332 g) with corrected transposition of the great arteries developed chronic lung disease due to a severe heart failure and post operative several complications. We applied intrapulmonary percussive ventilation (IPV) to the patient. IPV improved oxygenation concomitant with the improvement of respiratory condition and chest X-ray finding. However, the patient suffered from upper gastrointestinal bleeding 15 days after initiation of IPV therapy. The bleeding was healed several days after temporal termination of IPV, but recurred with resuming IPV therapy. The patient was irritable throughout the IPV therapy, and thus gastrointestinal bleeding of the patient could be due to stress induced by IPV therapy. IPV may be useful for the management of respiratory disturbance, often observed in low birth weight patients with congenital heart defects. However, gastrointestinal bleeding may occur and should be considered as a possible complication associated with IPV therapy. PMID- 19280952 TI - [Sweet's syndrome after off-pump coronary artery bypass grafting]. AB - We report a 73-year-old man with myelodysplastic syndrome (MDS) who developed Sweet's disease after off-pump coronary artery bypass grafting (OPCAB). Coronary angiography showed severe 3 vessel disease. We conducted OPCAB on triple vessels using bilateral internal thoracic artery and saphenous vein grafts. He had a high fever and rubor around the median skin incision on 5 postoperative day. Skin biopsy demonstrated acute febrile neutrophilic dermatitis (Sweet's disease). The cutaneous lesions improved gradually by corticosteroids. A careful attention should be paid postoperatively in a patient with hematological disorders such as MDS, aplastic anemia, and malignancy. PMID- 19280953 TI - [Candida mediastinitis after double closure technique for repairing ventricular septal perforation]. AB - A 69-year-old man was referred to our hospital due to acute myocardial infarction. Systolic heart murmur was first noted on the 23rd day after the onset, but no cardiac shunt flow was detected by echocardiography at that time. Six days later, cardiac function deteriorated rapidly, followed by oliguria and shock. Re-do echocardiography showed ventricular septal perforation. Emergency operation was performed, and septal perforation was seen on the anterior portion of the septum. In addition to infarct-exclusion-technique (Komeda-David method) with the equine pericardial patch, direct closure of the septal defect was performed (double closure technique). Fibrin glue was applied between the ventricular septum and the patch. After surgery, he suffered from Candida mediastinitis and received omentum plombage. Furthermore tracheotomy was performed for pneumonia. He recovered gradually, and was discharged about 3 months after surgery. Echocardiography showed no residual shunt. PMID- 19280954 TI - [Aortic valve repair and patch closure for unruptured aneurysm of non-coronary sinus of valsalva with aortic regurgitation]. AB - The patient was a 53-year-old woman who complained of chest pain. Echocardiography and angiography revealed mild aortic regurgitation (AR) with an eccentric jet and an unruptured aneurysm of the non-coronary sinus of Valsalva which protruded into the left atrium. Operative findings showed that tethering due to elongation of the circumference of the aortic wall at the level of the non coronary sinus commissures caused AR. Then patch closure and partial sino-tubular (ST) junction plication were carried out. Postoperative echocardiography showed decrease of AR and complete repair of the aneurysm of Valsalva sinus. PMID- 19280955 TI - [Long term follow-up of a case of pulmonary epithelioid hemangioendothelioma]. AB - A 54-year-old woman was referred to our hospital for bilateral multiple nodular shadows on a medical checkup chest X-ray film. Computed tomography also showed multiple small nodules in bilateral lung fields. Thoracoscopic lung biopsy was performed. The tumor cells showed immunohistochemically positive staining for factor VIII-related antigen, CD 34, and vimentin. The tumor therefore was diagnosed as pulmonary epithelioid hemangioendothelioma. The patient had been followed without any treatment, and died 12-years and 4 months after the diagnosis. PMID- 19280956 TI - [Multilocular thymic cyst]. AB - We describe a case of multilocular thymic cyst with severe acute inflammation. A 23-year-old man was admitted to our hospital with a sudden onset of chest pain and high fever. A computed tomography scan showed multilocular cystic lesion at anterior mediastinum. We resected the tumor with the thymus by median sternotomy. Macroscopically, the mediastinal mass showed thick-walled multiloculated cavities filled with turbid yellow fluid. Histological examination revealed that the tumor was a multilocular thymic cyst which is reported by Suster. PMID- 19280957 TI - [Bilateral pneumothorax of an amyotrophic lateral sclerosis patient under mechanical ventilation]. AB - Amyotrophic lateral sclerosis (ALS) is a progressive and fatal disease. To support breathing of some patients with ALS in its severe condition, mechanical ventilation is indispensable. However, mechanical ventilation has been known to induce pneumothorax by the damage of lung cells in response to mechanical stretch. An ALS 50-year-old male on mechanical ventilation was referred to our department for left pneumothorax. After an unsuccessful drainage for a couple of weeks, he underwent a partial resection of the left lung. On 3rd postoperative day (POD) the left lung collapsed again. Moreover, on 6th POD, the right pneumothorax occurred. Regarding the right pneumothorax, drainage was effective with a continuous pressure of -10 cm H2O, and the chest tube was removed soon. An air leak from the left chest tube persisted, and the left lung expansion was not enough with its apex line around the clavicle. On 42nd POD, a drainage pressure was increased up to -15 cm H2O. Then an air leak disappeared, and the lung expansion was obtained. The adjustment of a chest tube drainage pressure seems to be important, especially when a pneumothorax patient on mechanical ventilation is treated. PMID- 19280958 TI - [Surgical removal of native aortic valve thrombosis associated with acute myocardial infarction and protein C deficiency; report of a case]. AB - Native aortic valve thrombosis is an uncommon event. We describe the case of a 76 year-old man who suffered acute myocardial infarction associated with native aortic valve thrombosis diagnosed by cardiac catheterization. Since the thrombus was localized on the right coronary cusp and occluded right coronary artery, rescue revascularization was performed using perfusion catheter, which was continuously engaged to the right coronary artery. Operation was immediately performed under cardiopulmonary bypass. After incision of ascending aorta, thrombus was removed easily and aortic valve was preserved without degenerative change. Histological study showed a typical thrombus without any specific findings. He had a good clinical course and discharged 9 days after the operation. He had no history of heart valve disease, left heart catheterization or bacterial endocarditis. Since laboratory data showed 41% in protein C antigen and 32% of protein C activity, he was diagnosed of protein C deficiency. Our report emphasize that this thrombus formation may be caused by protein C deficiency. PMID- 19280959 TI - [Perforation due to new myocardial infarction long time; report of a case]. AB - A 65-year-old man was admitted to the hospital because of effort chest pain, 8 years after he received coronary artery bypass grafting [CABG: left internal thoracic artery (LITA) to left anterior descending artery (LAD), saphenous vein graft (SVG) to first diagonal branch (D1) and SVG to postero-lateral branch (PL)]. Emergent coronary angiography revealed right coronary artery occlusion with well patent bypass grafts. Percutaneus coronary intervention (PCI) was performed successfully, but 9 hours later, ventricular septal perforation (VSP) was occurred. Swan-Ganz catheter revealed that pulmonary to systemic blood flow ratio (Qp/Qs) was 2.6. In spite of intensive medical care, his hemodynamics was gradually exacerbated. Subsequent intracavitary repair with equine pericardial patch, sutured using interrupted mattress sutures with felt pledgets, was performed. He had an uneventful recovery thereafter, extubated and weaned from an intra-aortic balloon pumping at the 1st day. He was discharged from hospital on the 27th postoperative day. PMID- 19280960 TI - [Aortoesophageal fistula due to thoracic aortic aneurysm; report of a case]. AB - We report a 74-year-old man with aortoesophageal fistula due to aotic aneurysm. He underwent 2 stage operations. At the 1st operation the graft replacement of thoracic aorta and esophagectomy were performed. Inflammatory reactions improved with systemic administration of antibiotics and continuous irrigation of the thoracic cavity. On the 21st postoperative day, the esophagus was reconstructed by gastrointestinal interposition technique via ante-thoracic route. On the 58th post operative day he was discharged. PMID- 19280961 TI - [Surgical experience of idiopathic peripheral pulmonary arterial aneurysm; report of a case]. AB - Peripheral pulmonary arterial aneurysm has rarely been reported, and although most cases show symptoms of hemoptysis. We experienced a case of idiopathic peripheral pulmonary arterial aneurysm in the asymptomatic period noted on medical examination. The patient was a 49-year-old man with coin lesion which was 15 mm in diameter in the right lower lung field on chest X-ray film at the 1st visit. It was strongly stained on enhanced computed tomography in the right S4 and continuity with the pulmonary arteries was suggested. The right middle lobectomy was performed under the diagnosis of peripheral pulmonary arterial aneurysm. Histological examination of the resected specimen revealed that the prominent thinning in the arterial wall, however, there was no evidence of specific chronic inflammation. The postoperative course was good. The patient has been well without new pulmonary arterial aneurysm for 2 years after the surgery. PMID- 19280962 TI - [Mediastinal lymph node carcinoma of unknown primary site; report of a case]. AB - A 44-year-old woman was admitted to our hospital because of mediastinal mass. Serum levels of carcinoembryonic antigen (CEA) were found to be elevated. No preoperative examination could detect the primary lesion. The tumor was resected through right thoracotomy. Histological examination revealed poorly differentiated adenocarcinoma. She was diagonosed as metastatic mediastinal lymph node carcinoma of unknown primary site. She received radiotherapy and chemotherapy and is desease free 29 months after operation. Good results may be obtained by multimodality therapies for cancer in mediastinal lymph node of unknown primary site. PMID- 19280963 TI - Next generation calcium phosphate-based biomaterials. AB - It has been close to a century since calcium phosphate materials were first used as bone graft substitutes. Numerous studies conducted in the last two decades have produced a wealth of information on the chemistry, in vitro properties, and biological characteristics of granular calcium phosphates and calcium phosphate cement biomaterials. An in depth analysis of several key areas of calcium phosphate cement properties is presented with the aim of developing strategies that could lead to break-through improvements in the functional efficacies of these materials. PMID- 19280964 TI - Bio-active restorative materials with antibacterial effects: new dimension of innovation in restorative dentistry. AB - Restorative materials in the new era should be "bio-active", and antibacterial effects are highlighted as one of the important properties. In order to achieve resin-based restoratives with antibacterial effects, an antibacterial monomer MDPB has been developed. The primer incorporating MDPB demonstrated cavity disinfecting effects, and the world's first antibacterial adhesive system employing the MDPB-containing primer was successfully commercialized. MDPB is potentially applicable to various restoratives since immobilization of the antibacterial component by polymerization of MDPB enables no deterioration in mechanical properties of cured resins and exhibition of inhibitory effects against bacterial growth on their surfaces. For glass-ionomer cements used for atraumatic restorative treatment, the approach to provide antibacterial activity has been attempted by addition of chlorhexidine. Incorporation of 1% chlorhexidine diacetate was found to be optimal to give appropriate antibacterial and physical properties, being effective to reduce the bacteria in affected and infected dentin in vivo. PMID- 19280965 TI - Electrochemical surface modification of titanium in dentistry. AB - Titanium and its alloys have good biocompatibility with body cells and tissues and are widely used for implant applications. However, clinical procedures place more stringent and tough requirements on the titanium surface necessitating artificial surface treatments. Among the many methods of titanium surface modification, electrochemical techniques are simple and cheap. Anodic oxidation is the anodic electrochemical technique while electrophoretic and cathodic depositions are the cathodic electrochemical techniques. By anodic oxidation it is possible to obtain desired roughness, porosity and chemical composition of the oxide. Anodic oxidation at high voltages can improve the crystallinity of the oxide. The chief advantage of this technique is doping of the coating of the bath constituents and incorporation of these elements improves the properties of the oxide. Electrophoretic deposition uses hydroxyapatite (HA) powders dispersed in a suitable solvent at a particular pH. Under these operating conditions these particles acquire positive charge and coatings are obtained on the cathodic titanium by applying an external electric field. These coatings require a post sintering treatment to improve the coating properties. Cathodic deposition is another type of electrochemical method where HA is formed in situ from an electrolyte containing calcium and phosphate ions. It is also possible to alter structure and/or chemistry of the obtained deposit. Nano-grained HA has higher surface energy and greater biological activity and therefore emphasis is being laid to produce these coatings by cathodic deposition. PMID- 19280966 TI - Smart materials in dentistry--future prospects. PMID- 19280967 TI - A review of dental CAD/CAM: current status and future perspectives from 20 years of experience. AB - In this article, we review the recent history of the development of dental CAD/CAM systems for the fabrication of crowns and fixed partial dentures (FPDs), based on our 20 years of experience in this field. The current status of commercial dental CAD/CAM systems developed around the world is evaluated, with particular focus on the field of ceramic crowns and FPDs. Finally, we discuss the future perspectives applicable to dental CAD/CAM. The use of dental CAD/CAM systems is promising not only in the field of crowns and FPDs but also in other fields of dentistry, even if the contribution is presently limited. CAD/CAM technology will contribute to patients' health and QOL in the aging society. PMID- 19280968 TI - International standards: why do we need them? PMID- 19280969 TI - Shear bond strengths of different adhesive systems to white mineral trioxide aggregate. AB - This study was conducted to compare the shear bond strengths of different adhesive systems to White Mineral Trioxide Aggregate (WMTA). To this end, 40 cylindrical acrylic blocks with a hole in the middle measuring 4 mm diameter x 2 mm height were prepared. The hole was filled with WMTA, and the specimens were allocated into four groups: Group 1--Prime & Bond NT; Group 2--AdheSE; Group 3- Xeno III; and Group 4--Adper Prompt L-Pop. In each group, a different adhesive system and a compomer (Dyract AP) were applied over WMTA. Shear bond strengths were measured using a universal testing machine, and then the data were subjected to one-way ANOVA and Scheffe's post hoc test (p<0.05). Significantly lowest shear bond strength value was obtained with Adper Prompt L-Pop, whereas Prime & Bond NT revealed higher shear bond strength than the others (p<0.05). In conclusion, an etch-and-rinse adhesive system--which exhibited significantly higher shear bond strength than self-etch adhesive systems--could be a preferred choice when placing compomer materials upon WMTA. PMID- 19280970 TI - Chemical mechanical polishing of titanium with colloidal silica containing hydrogen peroxide--mirror polishing and surface properties. AB - Chemical mechanical polishing (CMP) of cpTi (Ti) was carried out using two types of slurries, acidic and basic colloidal silica containing H2O2 up to 3 wt%, to obtain flat and mirror-polished surfaces without any contaminated and reacted layers. Polishing behavior and surface properties were investigated using AFM, EPMA, and XPS. Weight loss of Ti polished by CMP using the basic slurry was larger than that using the acidic one, and surface roughness was less than 2 nm RMS when basic slurry containing 3 wt% H2O2 was used. Moreover, three kinds of chemical species, OH-, O2-, and H2O, were detected on the Ti surfaces polished by CMP using these slurries. Results of this study showed that CMP using colloidal silica containing H2O2 successfully created a mirror-polished surface without contaminated and reacted layers. PMID- 19280971 TI - Application of haptic device to implant dentistry--accuracy verification of drilling into a pig bone. AB - To enable accurate implant placement and precise drilling following preoperative simulation, we developed the BoneNavi system. To realize more precise drilling when the holes are upsized, two methods of surgical guiding were attempted in the present study. One involved using multiple surgical guides with titanium tubes of different diameters, and the other involved using a single surgical guide but employing titanium drill guide tubes with different diameters. Drilling accuracy of the two newly developed methods was examined and compared with the results of drilling into a pig bone using only the initial surgical guide. Deviations of the position and angle with the two novel methods were similar: 0.17 mm and 1 degree respectively. As for the control group whereby drilling was done using only the initial surgical guide, the deviations were 0.25 mm and 3.50 degrees--which were significantly larger than those achieved with the two novel methods. In light of the results obtained, our newly developed BoneNavi system is especially applicable for severe clinical cases that require precise implant placement. PMID- 19280972 TI - Development of a multiwalled carbon nanotube coated collagen dish. AB - Carbon nanotubes (CNTs) are one of the most interesting nanomaterials because of their excellent characteristics. In this study, a transparent CNTs coating for cell culture dishes was developed and its properties for cell culture were estimated. Carboxylated multiwalled carbon nanotubes (MWCNTs) were dispersed in aqueous sodium cholate solution and applied on a collagen type I-coated cell culture dish (cover glass). The dish surface was homogeneously covered by MWCNTs without aggregation. The MWCNT-coated dish was slightly gray and had good transparency, so conventional optical microscopic observation of the cells on the MWCNT-coated dish was possible. Rat osteoblast-like cells cultured on the MWCNT coated dish showed slightly lower viability and proliferation compared to the collagen-coated dish. The cell adhesion on the MWCNT-coated dish was much higher than that on the collagen-coated dish. Therefore, MWCNT-coating for dishes will be a useful new material for cell culture. PMID- 19280973 TI - Effects of application time and agitation for bonding orthodontic brackets with two self-etching primer systems. AB - The purpose of this study was to evaluate the shear bond strength, adhesive remnant index (ARI) and etching effect of two self-etching systems (Transbond Plus, Beauty Ortho Bond) at different application times (3, 10, 30 seconds) with and without agitation. Share bond strengths were examined and the ARI score was estimated (n=15). Enamel surfaces after priming were examined by SEM. Increasing application time and applying agitation of self-etching primers produced slightly increased etching efficacy. Increasing application time did not significantly increase the shear bond strength. Also, agitation did not significantly increase the shear bond strength except for specimen bonded with the Beauty Ortho Bond with 10 seconds application time. No significant difference was found in bond failure sites among 10 groups. Although increasing application time and applying agitation of both self-etching primers did not affect the shear bond strength, increasing application time in indirect bonding technique should not cause remarkable adverse effect. PMID- 19280974 TI - Effectiveness of surface protection for glass-ionomer, resin-modified glass ionomer and polyacid-modified composite resins. AB - The purpose of this study was to evaluate the effectiveness of several surface protectors for a glass-ionomer, a resin-modified glass-ionomer, and a polyacid modified resin cement by determining dye uptake spectrophotometrically. 378 samples, made up of Ionofil U, Vitremer, and Dyract, were prepared and divided into groups of seven each. Positive and negative control specimens remained unprotected while the experimental specimens were protected with Finishing Gloss, Protect-It, LC Varnish, Adper Single Bond, or a nail varnish. The experimental groups and positive controls were immersed in 0.05% methylene blue solution, while the negative controls were immersed in deionized water. Results were evaluated using variance analysis. Of the Ionofil U group, Adper Single Bond exhibited the least effective surface coating among the materials tested, while the best surface protection was obtained with LC Varnish in the Dyract group. However, no statistically significant differences were observed in the Vitremer group. PMID- 19280975 TI - Examination of composite resins with electron microscopy, microhardness tester and energy dispersive X-ray microanalyzer. AB - This study was conducted to examine the ultrastructures of eight recently improved light-cure restorative composite resins with scanning and transmission electron microscopes (SEM and TEM). Additionally, Vickers hardness, volume/weight fraction of filler, and chemical composition were analyzed. Composite resins selected for evaluation were Beautifil II, Clearfil AP-X, Clearfil Majesty, Estelite sigma, Filtek Supreme, Filtek Z250, Solare, and Synergy. SEM and TEM images revealed a great diversity in ultrastructure, and Vickers hardness test showed significant differences amongst all the composite resins (except between Clearfil Majesty and Estelite sigma, and between Filtek Supreme and Filtek Z250). By means of EDX, similar elements such as C, O, and Si were detected, but the concentration was different in every composite resin. Results obtained in this study served to validate that the methods employed in this study SEM and TEM at high magnification--were useful in examining the ultrastructures of composite resins. It was also found that the ultrastructure, size of filler particles, volume/weight fraction of filler, and chemical composition of the composite resins had an effect on Vickers hardness. Given the great diversity of ultrastructures amongst the composite resins, which stemmed from the different revolutionary technologies used to manufacture them, further studies are warranted in the search of clinical applications that optimally match the differing properties of these materials. PMID- 19280976 TI - A structure-activity relationship study on the mechanisms of methacrylate-induced toxicity using NMR chemical shift of beta-carbon, RP-HPLC log P and semiempirical molecular descriptor. AB - To clarify the mechanism of methacrylate-induced toxicity, a total of 24 acrylates, methacrylates, and dimethacrylates were chosen for a structure activity relationship (SAR) study in terms of NMR chemical shifts, semiempirical molecular descriptors, and reverse phase (RP)-HPLC log P. Molecular descriptors as well as bulk, electronic, and energy descriptors were calculated using the PM3/CONFLEX method. A significant multiple linear regression equation for methacrylates in mice was denoted as log 1/LD50 (which was function [ (E(HOMO)+E(LUMO))/2, log P]). Besides, significant linear regression equations for methacrylates were denoted as log 1/ED50 in HeLa S3 and in HGF cells as function [E(HOMO) and/or log P]. Results showed that the 13C NMR chemical shift of beta-carbon for methacrylates was correlated with their E(HOMO). Findings of this study thus suggested that it might be possible to predict methacrylate induced toxicity using physicochemical properties. PMID- 19280977 TI - Effect of bending on the mechanical properties of gold wrought-wire clasps: a non linear finite element analysis. AB - The purpose of this study was to assess the effect of bending of dental gold alloy wires on the mechanical characteristics of wrought-wire clasps. We conducted a simulation of large deformation in straight wires by means of non linear finite element (FE) analysis. A bending force increased the principal tensile stress on the outer surface of the bending corner and the compressive stress on the inner surface of the bending corner to their maximum values. After unloading with springback, a residual tensile stress was produced on the inner surface. A gold alloy wire clasp exhibited a relatively greater flexibility with small permanent deformation after the clasp tip deflection as compared to previously reported data for Co-Cr wires; this suggests that it is suitable for periodontally compromised teeth. Wire clasps are more susceptible to failure as compared to straight wrought wires because of the residual stress produced during the bending process. PMID- 19280978 TI - [Philosophical basis of modern public health]. PMID- 19280979 TI - [Issues associated with evaluating the effectiveness of influenza vaccination by observational studies]. AB - Vaccination against influenza is recommended for subjects at high risk of severe complications if infected with the influenza virus. The effectiveness of influenza vaccination among the elderly has been evaluated in many observational studies but only in a few randomized control trials. However, observational studies have various issues. First, the outcomes should be of high specificity with low nondifferential misclassification, such as laboratory-confirmed influenza or high fever. Differential misclassification between vaccinated and unvaccinated participants may also be a complicating factor due to behavioral differences in the habits of participants with regard to visiting physicians or in the diagnostic processes of physicians. Therefore, preventing differential misclassification by equally precise observation in compared groups seems to be more important than the specificity of outcomes, even if this can be achieved only by techniques with low-specificity outcomes. In observational studies of the elderly, vaccinated elderly individuals may be at low risk of influenza infection or death because they are health conscious. Such confounding may lead to an overestimation of the effectiveness of influenza vaccination. To minimize bias and to adjust for confounders, the effectiveness of influenza vaccination should be compared taking into account the following groups/indicators: influenza season and pre-season, well-matched season and mismatched season between vaccine and circulating viruses, severe season and mild season, high-specificity outcomes and low-specificity outcomes. Low effectiveness is to be expected with the latter of each pair. If no difference in effectiveness is observed between the two conditions in each pair, residual confounding may be present and the true underlying risk factors should be sought. For example, elderly individuals with impaired functional status in addition to chronic diseases may be less able to access vaccination. Hence it is necessary to evaluate the effectiveness of vaccination among such elderly individuals and promote vaccination in those people. PMID- 19280980 TI - [Prevalence and related factors for allergic diseases in Saitama]. PMID- 19280981 TI - [Vaccination coverage of foreign children and their parents' views on immunization services in Gunma Prefecture, Japan]. AB - OBJECTIVES: The aim was to evaluate vaccination coverage among foreign children living in an urban area of Gunma Prefecture, Japan and to examine their parents' views concerning the local immunization services. METHODS: A total of 321 foreign children aged 6 to 18 years in five international schools participated in school health examinations and provided vaccination information. Among the parents, 304 completed a questionnaire on their views about the immunization services. Another questionnaire survey was conducted in nursery schools for parents of 4629 Japanese children aged 0 to 6 years. Of the total, 3811 (82.3%) responded, and 2911 questionnaires answered by the parents who had children aged 3 years and older were eligible for the analysis. RESULTS: The study found a vaccination coverage of 86.2% for diphtheria and tetanus toxoids and pertussis (DTP), 86.5% for poliovirus vaccine, and 87.7% for BCG among the foreign children. Of their parents, 84% were born in Brazil. One third of the foreign children vaccinated for DTP, poliovirus and/or BCG had received each vaccine in Japan, while the others children had been vaccinated in their home countries. Among 162 parents with children immunized in Japan, 77% received the necessary information about immunization services from the local municipal office, and 80% had no major problems. However, 15% felt the language barrier. Among the Japanese children, non-vaccination rates for DTP, poliovirus vaccine, and BCG were 18.5%, 9.9%, and 3.5%, respectively. Of the Japanese parents, 85% knew immunization schedules from the municipal office, and 51% asked for night-time and holiday vaccination sites. CONCLUSION: These results suggest that vaccination coverage for DTP, poliovirus vaccine or BCG is relatively high among foreign children living in Japan. To promote higher rates of vaccination for those residents, however, accessibility of the municipal consultation services in foreign languages should be im-proved. PMID- 19280982 TI - [Syndrome of intracavitary hypertension in a surgical clinic]. AB - The problem of intracavitary hypertension syndromes characterized by similar pathophysiological abnormalities and different etiological factors is investigated and discussed. Compartment syndrome is believed to be the extreme variant of the disease. Specific markers and methods of correction (including surgical intervention) of intracavitory hypertension in critical situations are considered. PMID- 19280983 TI - [On pathogenetic significance of activation of lipid peroxidation in the mechanisms of disturbance of blood rheologic properties in experimental intoxication induced by fraction FII of vaccinal EV strain of Y. pestis]. AB - Changes of blood viscosity were induced by parenteral administration of fraction II of vaccinal EV strain of Y. pestis ("murine" toxin) to mice at doses equivalent to LD50 and LD25. The study revealed correlation between lipid peroxidation activity, severity of autointoxication, and integrated indices of blood theologic properties. PMID- 19280984 TI - [Endoscopic subfascial dissection of perforating veins of lower legs in patients with venous ulcers]. AB - Prospective analysis of the results of endoscopic subfascial dissection of perforating veins of lower legs in 68 patients with venous ulcers is presented. Four patients developed postoperative wound infection. Mean duration of hospitalization was 4.5 days. The patients were able to resume routine daily activity within 8 days after surgery. Mean length of ulcer healing was 7.0 months. Complete healing required 3 months in 22.1% of the patients, 6 months in 38.2%, 9 months in 66.2%, and 1 year in 97.1%. Ulcers recurred in 2 patients. Mean clinical and disability scores 8 months after surgery were on an average very low. It is concluded that subfascial dissection of perforating veins of lower legs in patients with venous ulcers improves clinical symptoms, promotes healing of ulcers, produces minimal postoperative complications, and plays an important role in correction of concomitant pathology. PMID- 19280985 TI - [Nanosystems and targeted transport of medicinal preparations to the brain]. AB - Intense efforts in the last decades have resulted in numerous inventions to improve drug delivery to CNS. Some of them have significant potential for clinical applications. At the same time, the variety of these strategies is in itself indicative of great difficulties inherent in the transfer of therapeutic and imaging agents across the blood-brain barrier. Combination of several approaches, such as encapsulation of drugs in nanocontainers, incorporation in micellar nanostructures, the use of Pluronic "unimers" for inhibition of drug efflux in endothelial cells of brain capillaries, appears to be the most promising modality. The same refers to the methods of intracellular transport of drug suspensions developed recently. All these achievements provide a solid basis for the further improvement of drug pharmacokinetic properties, reduction of systemic toxicity, increase of therapeutic efficacy, and earlier diagnosis of CNS pathology. PMID- 19280986 TI - [Nitric oxide and coronary heart disease]. AB - The NO system plays an important role in pathogenesis of coronary heart disease. Disturbances of reperfusion associated with myocardial infarction and post infarction heart failure are shown to be related to reduced synthesis and biological availability of NO. Variations of activity of three NOS isoforms (eNOS, nNOS, and iNOS) in patients with ischemic heart disease are linked to altered expression of the respective genes. They correlate with the character and severity of myocardial dysfunction. Cardiotropic effects of the above isoenzymes depend on their subcellular localization, interaction between themselves and between NO and reactive oxygen species, disturbed NO/redox balance, and other factors. Enhanced NOS activity and NO level at the premorbid stage of coronary heart disease have important implications for the prevention and treatment of this condition. PMID- 19280987 TI - [Cloning and characterization of a large metagenomic DNA fragment containing glycosyl-hydrolase genes]. AB - The problem of search for and characterization of enzymes synthesized by non cultivated microorganisms is presently being settled by creating metagenomic libraries. A 6000-clone library with the average size of its inserts amounting to 15 bp has been constructed on the basis of total DNA isolated from cow rumen microorganisms. As the result of the screening of the library on plates with different substrates, a clone was selected that efficiently hydrolyzed lichenan and carboxymethylcellulose. The clone contained the recombinant plasmid pBlue-13 bearing a 12071 bp.-long metagenomic fragment carrying ten open reading frames, two of them being identified as glycosyl hydrolase genes. No homology of the metagenomic DNA with any known microorganism genomes was revealed. The amino acid sequence, deduced on the basis of frame 4 and denoted by Xyl3A, bears resemblance with beta-xylosidases of glycosyl hydrolase Family 3. Frame 6 encodes polypeptide Cel5A homologous to cellulases of glycosyl hydrolase Family 5. The amino acid sequences deduced on the basis of seven out of ten open reading frames were homologous to proteins of microorganisms belonging to the Bacteroides sp. family, the bacteria inhabiting mammalian intestines. PMID- 19280988 TI - [Inverse correlation between GC-content of bacterial genomes and the level of preterminal codons usage in them]. AB - The purpose of this work was to find the character of the dependence between the GC-content of the gene and the level of preterminal codons usage inside it. 84 codon usage tables were used as the material (each of them contains average frequencies of codon usage for all encoding districts belonging to one bacterial specie). We also used nucleotide sequences encoding for active centers of bacterial adenylate cyclases. Inverse correlation between the GC-content (G+C) and the total level of preterminal codons usage (PCU) was observed (R = - 0.97). For nucleotide sequences encoding for active centers from adenylate cyclases class I the coefficient of correlation between G+C and PCU is -0.75. For sequences encoding for active centers from adenylate cyclases class III the coefficient of correlation between G+C and PCU is -0.91. Preterminal codons are mostly GC-deficient relative to their synonymous nonpreterminal codons and in absolute terms. The cause of the inverse correlation between G+C and the level of preterminal codons usage is an increase in their frequencies of usage due to mutational AT-pressure and the decrease of their frequencies of usage due to mutational GC-pressure. The evidence of the frequent fixation of nonsense mutations in GC-deficient bacteria was found. The practical conclusion is the following: the higher is the GC-content of gene or genome, the lower is the probability of nonsense mutation inside it. PMID- 19280989 TI - [Research into the immunogenic properties of the recombinant cellulose-binding domain of Anaerocellum thermophilum in vivo]. AB - Development of new technology allows different antigens of a necessary degree of cleanliness to be obtained. This development is a major problem of modern medical biotechnology. A promising approach to this problem includes use of the affinity domains (tags) incorporated in structure of a recombinant antigen and capable to bind to corresponding sorbents. The method of preparation of ready-for-use injections containing complexes formed by soluble antigens on insoluble cellulose immunosorbent (not chemical conjugates) in one stage is based on the fusion protein technology. This approach includes preparation of two-component recombinant proteins containing an antigen of interest and the cellulose-binding domain (CBD), which spontaneously binds to cellulose containing sorbents with high binding constant. Research into the immunogenic properties of the CBD in the complex with cellulose and in the preparation of recombinant CBD in a rat model was performed. The titers of specific antibodies in rat serum induced by recombinant CBD and CBD in the complex with cellulose was evaluated. The CBD in the complex with cellulose was more immunogenic in comparison with CBD alone. The spectrum and levels of cytokines in collected rat serum induced by developed preparations was also measured using the microsphere-based Luminex Flowmetrix system (BioPlex). It was found that the amorphous cellulose was not an immunotolerant sorbent, because it induced the expression of the proinfammatory cytokines in vivo. PMID- 19280990 TI - [Recombinant pseudoadenovirus nanostructure with human lactoferrin gene: production and study of lactoferrin expression and properties in vivo usage of the construction]. AB - The Ad5-Lf pseudoadenovirus nanostructure (RPAN) was produced using homologous recombination in E. coli cells. This construction provided efficient expression of the Lf gene in permissive cell culture with high production rate of recombinant protein similar to native human Lf in some physical, chemical, and biological properties. Single intravenous injection of the construction into mice and rats was effective for prolonged production and circulation of recombinant human Lf in blood of experimental animals without toxic effects. The produced construction is promising for providing prolonged production of recombinant human Lf in the human body. PMID- 19280991 TI - [The study of the nonpathogenic influenza virus A/gull/Moscow/3100/2006 (H6N2) isolated in Moscow]. AB - The influenza virus A/gull/Moscow/3100/2006 (H6N2) was isolated from gull feces within the precincts of Moscow in autumn 2006. The nucleotide sequence of the complete genome (GenBank, EU152234-EU152241) and genotype (K, G, D, 6B, F, 2D, F, 1E) for this virus were determined. Phylogenetic analysis suggests that the H6N2 virus derived by numerous reassortment between viruses that have been circulating among different birds in Europe since 1999 and in South-East Asia (NA gene) for last years. Migratory birds probably introduced some of these viruses from South East Asia earlier. The strain A/gull/Moscow/3100/2006 is nonpathogenic for chicken embryos and mice and induces specific antibody production in mice. Similar to all avian influenza viruses A/gull/Moscow/3100/ 2006 it binds to Neu5Ac(2-3Gal receptors, but reveals higher affinity for fucosylated sialosugars (SLex) in contrast to the duck viruses, as was shown in receptor specificity assay and clarified due to modeling the accommodation of SLex into receptor binding site of duck and gull influenza virus hemagglutinin. PMID- 19280992 TI - Linkage of the Third National Health and Nutrition Examination Survey to air quality data. AB - OBJECTIVES: This report describes the linked data file obtained as a result of combining air pollution data and National Health and Nutrition Examination Survey (NHANES) III data. METHODS: Average annual air pollution exposures to particulate matter consisting of particles smaller than 10 micrometers in diameter (PM10), sulfur dioxide (SO2), nitrogen dioxide (NO2), and carbon monoxide (CO) were created for NHANES III examined persons by averaging values from monitors within a 5-, 10-, 15-, and 20-mile radius from the block-group centroid of their residence and in the county of their residence. Percentage records geocoded to block-group level, percentage records linked to air pollution, and distributions of exposure values were estimated for the total sample and various demographic groups. RESULTS: The percentages of respondents who were assigned countywide air pollution values ranges from a low of 43 percent in the case of NO2 data to a high of 68 percent in the case of PM10 data. Among the pollutants considered, PM10 data provides the best coverage. Of all the metrics created, the highest coverage is achieved by averaging readings of monitors located within a 20-mile distance from the centroid of respondents' block groups. Among the demographic variables analyzed, differences in air pollution coverage and exposure levels occur most often among groups defined by race and Hispanic origin, region, and county level of urbanization. However, differences among groups depend on the pollutant and geographic linkage method. The linked dataset provides researchers with opportunities to investigate the relationship between air pollution and various health outcomes. PMID- 19280994 TI - Metabolic syndrome: an entity that lacks a clear definition but which is useful to identify in clinical practice. PMID- 19280995 TI - Human paraoxonase gene polymorphisms and coronary artery disease risk. AB - BACKGROUND: Complex diseases such as coronary artery disease (CAD), hypertension and diabetes are usually caused by individual susceptibility to multiple genes, environmental factors, and the interaction between them. The paraoxonase 1 (PON1) enzyme has been implicated in the pathogenesis of atherosclerosis and CAD. Two common polymorphisms in the coding region of the PON1 gene, which lead to a glutamine (Q)/arginine (R) substitution at position 192 and a leucine (L)/methionine (M) substitution at position 55, influence PON1 activity. Studies have investigated the association between these polymorphisms and CAD, but with conflicting results. AIMS: 1) To evaluate the association between PON1 polymorphisms and CAD risk; and 2) to study the interaction between PON1 polymorphisms and others in different candidate genes. METHODS: We evaluated the risk of CAD associated with PON1 Q192R and L55M polymorphisms in 298 CAD patients and 298 healthy individuals. We then evaluated the risk associated with the interaction of the PON1 polymorphisms with ACE DD, ACE 8 GG and MTHFR 1298AA. Finally, using a logistic regression model, we evaluated which variables (genetic, biochemical and environmental) were linked significantly and independently with CAD. RESULTS: We found that the PON1 55MM genotype was more common in the CAD population, but this did not reach statistical significance as a risk factor for CAD, while PON1 192RR presented an 80% higher relative risk compared to the population without this polymorphism. The interaction between PON1 192RR and MTHFR 1298AA, sited in different genes, increased the risk for CAD, compared with the polymorphisms in isolation (OR=2.76; 95% CI=1.20-6.47; p=0.009), as did the association of PON1 192RR with ACE DD, which presented a 337% higher risk compared to the population without this polymorphic association (OR=4.37; 95% CI=1.47-13.87; p=0.002). Similarly, the association between PON1 192RR and ACE 8 GG was linked to an even higher risk (OR=6.23; 95% CI=1.67-27.37; p<0.001). After logistic regression, smoking, family history, fibrinogen, diabetes, Lp(a) and the association of PON1 192RR + ACE 8 GG remained in the regression model and proved to be significant and independent risk factors for CAD. In the regression model the latter association had OR=14.113; p=0.018. CONCLUSION: When analyzed separately, the PON1 192RR genotype presented a relative risk for CAD 80% higher than in the population without this genotype. Its association with other genetic polymorphisms sited in different genes, coding for different enzymes and belonging to different physiological systems, always increased the risk for CAD. After correction for other conventional and biochemical risk factors, the PON1 192RR + ACE 8 GG association remained a significant and independent risk factor for CAD. PMID- 19280996 TI - Genetic factors and cardiovascular disease. PMID- 19280997 TI - Stable angina and ST-segment depression: diagnostic value in patients undergoing coronary angiography. AB - BACKGROUND: Clinical history and exercise testing (ET) are the main methods used to screen for coronary artery disease (CAD). Correct interpretation of such data is essential in determining diagnostic and treatment strategies. OBJECTIVE: To assess the diagnostic value of stable angina (SA) and ET in patients with suspected CAD undergoing coronary angiography and to compare results between the sexes. METHODS: We assessed 62 patients undergoing coronary angiography; ET was performed in 54 of them, being considered positive for ischemia in cases of ST segment depression of > or =1 mm. The coronary angiograms were analyzed by two observers and patients were divided into two groups depending on the severity of CAD: Group 1: > or =70% stenosis (n=26), and Group 2: <70% stenosis (n=36). RESULTS: The patients' mean age was 58+/-9 years, and 55% were male. SA was found in 24% of cases and ET was positive for ischemia in 53%. SA showed a sensitivity of 46.1%, specificity 91.7%, positive predictive value (PPV) 80%, negative predictive value (NPV) 70%, positive likelihood ratio (LR+) 5.1 and negative likelihood ratio (LR-) 0.6, while the corresponding results for ET were sensitivity of 70.8%, specificity 60%, PPV 59%, NPV 72%, LR+ 1.75 and LR- 0.5. SA was shown to have a greater NPV in women, and the LR+ was also significantly higher in women. CONCLUSION: SA was shown to be a good diagnostic marker, with high specificity and PPV. The greater NPV and LR+ in women was shown to be superior for post-test prediction of angina. ET results were similar for both sexes and showed similar sensitivity and specificity to that reported in the literature. PMID- 19280998 TI - Some reflections on prehypertension. PMID- 19280999 TI - Transcatheter closure of a fenestration in intracardiac Fontan circulation. AB - We present a case of tricuspid atresia and the treatment that was used in a child now aged 12. The malformation was corrected at the age of five through intracardiac Fontan circulation. Later, the child developed a right-to-left shunt due to dehiscence in the intra-atrial conduit, causing significant desaturation. This was treated by transcatheter closure with a 10 mm Amplatzer device at 8 years of age. Currently the child is asymptomatic with no residual shunts, and the position of the device is normal. The percutaneous closure of Fontan fenestrations using the Amplatzer device is a therapeutic approach that provides good results. It results in normal oxygen saturation and has low morbidity. The technique is simple, available to most cardiac catheterization laboratories, safe, and offers a reasonable cost-benefit ratio. In addition, the design of the device is suitable for this type of defect. PMID- 19281000 TI - Aortopulmonary window. Experience of eleven cases. AB - Aortopulmonary (AP) window is a communication between the ascending aorta and the main pulmonary artery, in the presence of two separate arterial valves arising from separate subarterial ventricular outflow tracts. It is a rare anomaly that accounts for approximately 0.1% to 0.2% of all congenital heart disease. The purpose of this study was to assess clinical features, surgical treatment and outcome of patients with aortopulmonary window referred to a tertiary pediatric cardiac center over a 30-year period. Eleven patients were diagnosed with AP window, ten with the proximal type and one with the distal type defect. Age at first evaluation ranged from three days to 13 years (mean 44.5+/-63.3 months; median three months). Echocardiography enabled correct diagnosis in the four most recent cases. Seven patients underwent closure of the AP window through a transaortic approach and three patients underwent ligation. Simultaneously, significant associated cardiac anomalies were corrected in three patients: correction of interrupted aortic arch in two patients and closure of ventricular septal defect and Dacron patch enlargement of the right ventricular outflow tract in one patient. One patient was not operated because of fixed high pulmonary vascular resistance. Operative mortality was 10% (1/10) and there were no late deaths. Mean follow-up was 10+/-4.9 years. All nine surviving operated patients are asymptomatic, without medication, with no residual defects and without pulmonary hypertension. In conclusion, advances in diagnostic and surgical approaches to AP window in recent years have enabled earlier intervention with good outcome. PMID- 19281001 TI - Ablation of pulmonary veins with a single diagnostic and ablation catheter. PMID- 19281002 TI - Delayed enhancement in nonischemic cardiomyopathies [116]. PMID- 19281003 TI - [Strategy of prevention and management of intraoperative fractures of acetabulum during primary total hip arthroplasty]. AB - OBJECTIVE: To analyse the causes and resultes of intraoperative occurrence of an acetabular fracture in primary total hip arthroplasty and discuss propriate prevention and treatment of this problem. METHODS: Between July 2004 and December 2006, 326 primary total hip arthroplasties were retrospectively reviewed and found that eleven patients (eleven hips) had sustained an intraoperative acetabular fracture involving 7 male and 4 female,aged from 37 to 71 years (mean 54 years). Analyzed the causes of fractures, the anatomic location, treatment and outcome of the fractures. Acetabular component designs were categorized and each design was analyzed for fracture risk. RESULTS: Among 11 cases, the fractures occurrenced as setting the prosthesis in 6 cases, grinding and drilling in 3 cases, drawning hook in 2 cases. There were 5 cases of central type fracture, 3 of posterior-wall, 2 of anterior, 1 of posterior-upper. Nine of these patients (nine hips) had been followed-up for 12 to 29 months(mean 18 months). There were no heterotopic ossification, no dislocation, no deep venous embolism. All fractures were healing. CONCLUSION: Acetabular fracture during primary total hip arthroplasty is a complication of acetabular fixation without cement, which should be think highly of. In the present series, retention of a stable cup is associated with uneventful osseous in growth and excellent early-term outcomes. PMID- 19281004 TI - [Clinical observations on 222 patients with cervical periarthritis of shoulder]. PMID- 19281005 TI - [Intramuscular ganglion: a report of 2 cases]. PMID- 19281006 TI - [Clinical study on the early reconstruction of complex acetabular fractures]. AB - OBJECTIVE: To explore the best opportunity of structure reconstruction for complex acetabular fracture in order to provide the accurate time for clinical operative treatment. METHODS: Complex acetabular fracture patients were divided into experimental group (93 cases, 96 hips) and control group (98 cases, 101 hips) randomly according to the operative time. The operation of patients were done respectively at the 7th, 10th day after injury in experimental group and control group. The operative time, the excellent and good rate of reduction,the postoperative complications,the joint function (ache to walk,joint activity),the SF-36 were evaluated. RESULTS: (1) The operative time of experimental group was obviously shorter than control group according to different fracture classification. (2) According to the standard of Matta' joint function and X-ray, the experimental group was better than control group. (3) The excellent and good rate of reduction in experimental group was obviously higher than control group, according to CT scan before operation and after operation. CONCLUSION: The early structure reconstruction of complex acetabular fracture can obviously decrease operative time and complications, increase the rate of operative reduction. PMID- 19280993 TI - Metabolic syndrome in Portugal: prevalence and implications for cardiovascular risk--results from the VALSIM Study. AB - INTRODUCTION: The metabolic syndrome (MS) is a constellation of risk factors of metabolic origin that is associated with increased risk of type 2 diabetes mellitus (DM) and cardiovascular disease (CVD). Several regional studies have been conducted to determine its prevalence, but they are insufficient to determine the situation nationally or to characterize overall cardiovascular risk in Portugal. OBJECTIVE: To determine the prevalence of MS and each of its components in adult primary health care users in Portugal. METHODS: The VALSIM Study, involving 719 general practitioners (GPs), was performed in a primary care setting, based on stratified distribution and proportional to the population density of each region of mainland Portugal and the islands of Madeira and the Azores. The first two adult patients scheduled for an appointment on a given day were invited to participate, irrespective of the reason for the consultation. After informed consent was obtained, a questionnaire on sociodemographic, clinical and laboratory data was completed by the GP. A previous diagnosis of coronary artery disease (CAD), stroke or DM was identified by the GP based on medical records. A diagnosis of MS was defined according to the NCEP ATP III criteria. Logistic regression multivariate analysis was used to assess the risk of MS according to age, body mass index (BMI), waist circumference (WC) and region of residence for each gender, and to determine the association of CAD, stroke and DM with gender, age, BMI, WC, blood pressure and previous diagnosis of hypertension (HT), fasting glucose and previous diagnosis of DM. RESULTS: The study included 16,856 individuals (mean age 58.1+/-15.1 years, 18-96 years; 61.62% women). The prevalence of MS adjusted for gender, age and size of region was 27.5% and showed regional variations, being highest in the Alentejo (30.99%) and lowest in the Algarve (24.42%). MS was more common among women and increased with age, BMI and WC. Independent protective factors were residence in the Algarve (odds ratio [OR]: 0.78; 95% confidence interval [CI] 0.66-0.92 p=0.002) or in Lisbon and Tagus Valley (OR: 0.83, 95% CI 0.77-0.91, p<0.001), while residence in the Northern (OR: 1.11; 95% CI 1.01-1.21, p=0.03) or Central regions (OR: 1.08; 95% CI 1.002-1.16, p=0.045) was an independent risk factor after correction for gender and age. MS was linked to increased prevalence of HT (OR: 3.88; 95% CI 3.61-4.18, p<0.001), and high blood pressure was the most frequent MS component, particularly in men (93.7%). MS was the most powerful factor associated with a diagnosis of DM, particularly in women (OR 7.23; 95% CI 6.22 8.40, p<0.001). Although there was a strong association between MS and CAD (OR: 1.16; 95% CI 1.01-1.34, p=0.043), the most potent risk factor associated with CVD was HT. CONCLUSIONS: The prevalence of MS in Portugal is high (27.5%) and is strongly linked to the occurrence of CVD, and in particular to DM. These results highlight the need to implement preventive strategies for reducing overall cardiovascular risk in the Portuguese population. PMID- 19281007 TI - [Antibiotic-PMMA beads combined with external fixator for treating the infected fracture nonunion]. AB - OBJECTIVE: To investigate the effects of antibiotic-PMMA (polymethyl methacrylate) beads combined with external fixator in treatment of infected fracture nonunion. METHODS: Twenty-two cases of infected fracture-nonunions were reviewed involving 20 male and 2 female with an average age of 34.68 years (ranging 21 to 74 years). The data consisted of 9 cases of tibial fractures, 2 distal fractures of the femur, 6 femoral shaft fractures, 3 intertrochanteric fracture of the femur and 2 humeral shaft fractures. The procedure included thorough debridement to wipe out dead bone and granulation tissue, then antibiotic-PMMA bead chains imbedded into the dead space. One week later, secondary debridement was performed, antibiotic-PMMA bead chains were changed according to result of bacterial culture and susceptibility test, and fractures were stabilized with external fixator. Three months after debridement, antibiotic PMMA bead chains were taken out and bone graft with autogenous iliac cancellous bone chips was performed. RESULTS: The mean follow-up period was 19.98 months (ranging 15 to 28 months). Infection was controlled in 20 cases. One tibial fracture and 1 intertrochanteric fracture of the femur needed repeated debridement 2 and 3 months after bone grafting respectively,because of infection recurrence and sinus formation. All 22 cases achieved bony union averaged 15.09 weeks after bone grafting with a range of 8 to 24 weeks. CONCLUSION: Thorough debridement, imbedding antibiotic-PMMA bead chains combined with external fixator and staged bone grafting has proven to be effective and simple for treatment of infected fracture nonunion. The antibiotic bead delivers high tissue levels,obliterates dead space, aids bone repair. PMID- 19281008 TI - [Methods selection for femoral neck fractures in old people]. PMID- 19281009 TI - [Some concentrated items in manipulative reduction dealing with the unsteady of atlantoaxial joint]. PMID- 19281010 TI - [Postoperative complications of acromioclavicular joint dislocation of Tossy III]. AB - OBJECTIVE: To analyze retrospectively the postoperative complications of acromioclavicular joint dislocation (Tossy III) and explore the preventative methods for the complications. METHODS: Forty-eight cases of acromioclavicular joint dislocation (Tossy III) were reviewed, 14 cases treated with open reduction and Kirschner wire combined with steel wire fixation (group A), 11 cases treated with open reduction and lag screw or steel wires fixation (group B), 23 cases treated with open reduction and AO clavicular hook plate fixation (group C). The acromioclavicular ligament, articular capsule, coracoclavicular ligament and coracoacromial ligament were repaired in all patients. The reasons of postoperative complications were analyzed. RESULTS: Forty-eight patients got average follow-up of 18 months. In group A, 8 patients obtained excellent results, 4 good and 2 poor; in group B, 7, 3 and 1, respectively; in group C, 21, 1 and 1, respectively. The excellence rate showed statistical difference between group A and C (P < 0.05). In group A, 4 cases with postoperative pain, 3 cases with periarthritis of shoulder, 3 cases with arthritides of acromioclavicular joint, 4 cases with internal fixation failure and 2 cases with recurrence of joint dislocation; in group B, 3, 2, 1, 3 and 1, respectively; in group C, 22, 2, 1, 2 and 1, respectively. There was no significant difference in postoperative complications in three groups (P > 0.05). CONCLUSION: Selection of the suitable internal fixation and reconstruction of coracoclavicular and coracoacromia ligaments is the basic operation. Acromioclavicular space debridement, ligamentous reconstruction, rigid internal fixation are effective methods to reduce postoperative complications for the treatment of acromioclavicular joint dislocation. PMID- 19281011 TI - [Analysis on complications of elastic nail treating children's long bone fractures]. AB - OBJECTIVE: To analyze the reasons on complications of treatment with elastic nail in children's long bone fracture. METHODS: Sixty-six cases (75 parts of long bone fratures) were treated by elastic nail including 49 male and 17 female. The age ranged from 3 to 17 years, mean 7.8 years. There were 35 femur fractures (2 cases were hibateral), 20 tibia and fibula fractures (12 cases were tibia fractures), 8 radial fractures (1 case was ulna fracture) and 3 humerus fractures. The cases included 4 open fractures and 62 closed fractures. All cases were fresh fractures, no multi-segmental fractures. Three cases associated with brain and chest injuries. These cases were treated by open or closed reduction and internal fixaion with elastic nail. A cast or brace had been used after operation for a month. Following-up included the function of the joint,the bottom of the nail and the callus. Complications were timely recorded. RESULTS: All the patients were followed-up for 12 to 29 months, averaged 17 months. The cases occurrenced compilications including 2 cases of nonunion, 2 of new fracture, 1 of displacment, 4 of joint dysfunction, 3 of irritation of the bottom of the nail and 1 malunion. CONCLUSION: Strict indication, well design,canonical operation is a good way to avoid compliacations. At the same time,early treatment can reduce the sequela. PMID- 19281012 TI - [Clinical observation on external fixator for treating calcaneal fractures]. PMID- 19281013 TI - [Clinical observation on external humeral epicondylitis treated with back rotation traction]. AB - OBJECTIVE: To evaluate the clinical effect of manipulation on external humeral epicondylitis, and to explore the functional mechanism and ideal treatment. METHODS: Eighty-six patients who had been treated with acupuncture, obturation and needle-knife were divided into routine group and treatment group randomly. In routine group, there were 42 cases (male 13, female 29, means 40.8 years); and in treatment group there were 44 cases (male 16, female 28, means 41.2 years). There's no further treatment for the routine group after the therapy above, while the treatment group was added with back-rotation traction manipulation. RESULTS: Taking Verhaar therapy effect appraisal system of tennis-ball elbow to evaluate elbow function. After 7 days of therapy, the results were excellent in 13 cases, good in 16, fair in 4, poor in 9 in the routine group; and excellent in 38, good in 4 and fair in 2 in treatment group; and the effect in the treatment group were better than that of the routine group (P < 0.010). Half a year later, in the routine group 38 cases recurrenced and in the treatment group 10 cases recurrenced. CONCLUSION: Making manipulation after routine acupuncture, local obturation and needle-knife has active meaning to remove trauma inflammation, prevent re-conglutination, promote recovery and prevent recurrence. PMID- 19281014 TI - [Effects of supracondylar fractures of humerus treated by three different surgical methods]. AB - OBJECTIVES: To research the effects of supracondylar fractures of humerus treated by three different surgical methods. METHODS: There were 103 patients with supracondylar fractures of humerus including 58 male and 45 female. The age was from 3 to 12 years old with the average of 7.4 years. To divide the cases into three groups by the operative approach in the humerus, the inside approach were in 37 cases (group A), the outside in 35 cases (group B) and the rear in 31 cases (group C). Comparing the reduction of the stretching and bending function of elbow joint and incidence of elbow varus. RESULTS: All patients were followed-up for 1.5 to 8 years after operation with the average of 32 months. The reduction of stretching and bending function of elbow joint occurrenced in 6 cases in group A, 6 cases in group B and 25 cases in group C, the incidenc of group C were obvious more than group A and B (P < 0.01). The elbow varus occurred in 8 cases in group A, 17 cases in group B and 7 cases in group C, the incidence of group B were obviously more than group A and C (P < 0.05). CONCLUSION: There are different effects between three kinds of operative approaches in the humerus. The incidence of outside group is higher on the varus of elbow. The incidence of rear group is higher on the ability of elbow joint stretching and bending. The effect of inside group is sure on the operative approach in the humerus. PMID- 19281015 TI - [Design and application of the instruments for implantation of transarticular screws in low cervical spine]. AB - OBJECTIVE: To design and apply the instruments for implantation of transarticular screws in low cervical spine and to evaluate the value. METHODS: Two cadavers were used to explore the value of the instruments. Seventeen patients who suffered from cervical diseases were also included in the group,sixty-eight transarticular screws inserted by use of the instruments. RESULTS: All transarticular screws were implanted ease in the low cervical spine without split of facets,guided by the instruments. All patients were followed up three to twelve months. Neither neurologic deficit nor failure of internal fixation was found due to implantation of transarticular screws. CONCLUSION: Transarticular screws can be inserted safely and smoothly guided by the instruments, which have values to be introduced widely. PMID- 19281016 TI - [Skin flap with a branch of the tibial posterior artery for repairing the skin defect of foot and ankle]. PMID- 19281017 TI - [Modified Kocher procedure for the treatment of shoulder dislocation]. PMID- 19281018 TI - [Research on the role of metabolism of fatty substance and osteoclast activity during the development of steroid-induced necrosis of femoral head]. AB - OBJECTIVE: To explore the role of metabolism of fatty substance and osteoclast activity during the process of steroid-induced necrosis of femoral head through mice model inducing and model index measurement. METHODS: Forty SD male mice were divided into 2 groups randomly, the control group and the experiment group. After the gluteal injection of colibacillus endotoxin,the experiment group was given gluteal injection of prednisolone acetate 35.5 mg/kg per week, and 2 ml of normal saline to the control group. The mice were killed 12 weeks later and tested the content of Trap-5b, TC and TG of the blood serum. Vitodynamics, bone density were measured and sections of HE staining, Ca2+ and TRACP staining was made then statistic analysis was performanced. RESULTS: The content of TC, TG and Trap-5b increased apparently (P < 0.01). Large amount of osteoclasts were found in local medullary cavity. There was severe bone loss and decrease of vitodynamics in subchondral bone (P < 0.01) in experiment group. CONCLUSION: Metabolic disorder of fatty substance is the key pathogenesis of steroid-induced avascular necrosis of femoral head. Decrease of vitodynamics in subchondral bone due to hyperactivity and increase of osteoclast lead to collapse of femoral head directly. PMID- 19281019 TI - [Treatment of ankle lateral collateral ligament injury and fractures of the bottom of the fifth metatarsal with herbal compress and the reform of the U shaped gypsum]. PMID- 19281020 TI - [Effects of yougui drink on the osteogenic differentiation of mesenchymal stem cells in the proximal femur in steroid-induced avascular necrosis of femoral head]. AB - OBJECTIVE: To investigate the effects of Yougui drink on osteogenic differentiation of bone mesenchymal stem cells (BMSCs) in the proximal femur in steroid-induced avascular necrosis of femoral head in vitro. METHODS: The BMSCs were induced to osteoblasts in vitro with a conditional medium. The cells cultured with Yougui drink serum were compared with the ordinary one, which both were examined by microscopy, MTU method, ALP and calcium node. RESULTS: Microscopy and MTU studies showed that Yougui drink could improve proliferation of the osteoblasts. ALP and calcium node studies showed that Yougui drink could improve the activity of the osteoblasts (P < 0.01). CONCLUSION: Yougui drink has positive effects on osteogenic differentiation of mesenchymal stem cells in the proximal femur in steroid-induced avascular necrosis of femoral head in vitro. PMID- 19281021 TI - [Diagnosis and treatment of traumatic subscapularis tendon ruptur]. PMID- 19281022 TI - [Effects of Gukang on bone-source alkaline phosphatase (BALP) and insulin-like growth factor-1 (IGF-1) in serum of spaying rats]. AB - OBJECTIVE: To investigate the effects of Gukang on bone-source alkaline phosphatase (BALP) and insulin-like growth factor 1 (IGF-1) in serum of spaying rats and the mechanism of curative effect of Gukang on osteoporosis. METHODS: Sixty-eight 6-month-old SD rats were chosen and randomly divided into blank control group (22 rats with sham operation) and operation group (46 rats with spaying operation). Three months after operation, 10 rats were randomly chosen from each group and tested with bone mineral density in order to determine models of osteoporosis made. After modeling, operation group was divided into 3 sub groups: operation model group, estrogen group and Gukang group, 12 rars in each group. Twelve rats remained in blank control group. Every group were treated through intragastric administration therapy (volume 10 ml/kg). Blank control group and operation model group were irrigated with distilled water,estrogen group with estrogen and Gukang group with Gukang. Three months after treatment, serum of all groups were collected and tested for E2, BALP and IGF-1 with ELISA. RESULTS: The concentration of serum E2, BALP in estrogen group and Gukang group were higher than operation model group, there were significant difference (P < 0.05), but no significant difference in serum E2 between estrogen group and Gukang group (P > 0.05). The concentration of serum IGF-1 in Gukang group was higher than operation model group and blank control group, there were significant difference (P < 0.05). CONCLUSION: Gukang can increase the level of E2, BALP and IGF-1 in serum of spaying rats. Thus, it can indirectly promote reproduction of osteoblasts, inhibit activity of osteoclasts and promote bone formation. PMID- 19281023 TI - [Protective effect of glial cell line-derived neurotrophic factor infused into the tube setted into cavitas subarachnoidealis on spinal front corner motor neurons]. AB - OBJECTIVE: To investigate the effect of exogenous glial cell line-derived neurotrophic factor (GDNF) infused into the cavitas subarachnoidealis on cornu anterius medullae spinalis motor neurons after sciatic nerve axotomy. METHODS: Forty-eight healthy SD rats were divided into 2 groups randomly: GDNF group and NS group. The left sciatic nerve in rats were cut off. And then 0.9% saline (6 microl) and GDNF solution (6 microl) were injected into cavitas subarachnoidealis at L4-L6 in NS group and GDNF group,respectively. The rats were sacrificed on postoperative 1, 2, 4 and 8 weeks respectively. Their specimen of L4-L6 spinal cord were taken at different time and sectioned. The HE staining, Nissl staining and cholinesterase (ChE) staining in motor neurons were used for counting of motor neurons. RESULTS: In GDNF group the number of motor neurons in cornu anterius medullae spinalis and the ChE activity were higher than that of NS group. CONCLUSION: The exogenous GDNF infused into the cavitas subarachnoidealis are supposed to protect the degenerated spinal motor neuron from death after sciatic nerve injury. PMID- 19281024 TI - [Clinical application of spiral CT three-dimensional (3D) reconstruction in early diagnosis of fracture nonunion]. PMID- 19281025 TI - [Study the degree of cervical spinal canal stenosis by MRI in flexion and extension of the cervical vertebrae]. AB - OBJECTIVE: To study the degree and changes of cervical spinal canal stenosis by MRI scans in flexion and extension of the cervical vertebrae. METHODS: Thirty cases of cervical stenosis included 13 male and 17 female with an average age of 39 years ranging from 28 to 66 years. The sagittal diameter of cervical spinal canal were below 10 mm (absolute stenosis) in 12 cases,within 10 to 12 mm (correspondence stenosis) in 18 cases. MRI scans in neutrality, flexion, extension performanced and the degree of cervical spinal canal stenosis and the changes of spinal cord compression were evaluated after MRI scans obtained. RESULTS: Nineteen patients of extension occurrenced stenosis more serious, 8 patients of flexion occurrenced (P < 0.05). CONCLUSION: For the cervical stenosis imaging diagnostic, flexion and extension of cervical MRI scan can be used to supplement conventional MRI examination neutral position, and the extension of MRI is more sensitivity than neutral position and flexion bit. PMID- 19281026 TI - [Treatment of conmminuted patellar fractures with internal Ni-Ti patellar concentrator and tension bind wire fixation]. AB - OBJECTIVE: To study the clinical efficacy of the treatment of comminuted patellar fractures with internal NiTi-Patellar concentrator and tension bind wire fixation. METHODS: From March 2004 to June 2007, 38 cases of fresh comminuted patellar fractures were treated with internal NiTi-Patellar concentrator and tension bind wire fixation. There were 25 males and 13 females,ranging from 21 to 64 years (mean 42.5 years). All were comminuted fractures with displacement, 16 cases were 3 fragments, 14 cases were 4 fragments, 8 cases were 5 fragments. There were other fractures in 8 cases. During followed-up, knee function and complications were evaluated. RESULTS: All patients were followed up for 8 to 24 months (mean 15 months) and obtained complete bone union. No case of implant was loosening and fragment displacement, traumatic arthritis occured in 2 cases. Under Lysholm & Gillquist score, the results were excellent in 17 cases, good in 19, fair in 2. CONCLUSION: Internal Ni-Ti-Patellar concentrator and tension bind wire fixation is one of the ideal methods for the treatment of comminuted patellar fracture, which could provide satisfied reduction, reliable fixation and good functional recovery. PMID- 19281027 TI - [Clinical treatment of acute traumatic central cervical spinal cord syndrome without fracture and dislocation]. AB - OBJECTIVE: To analyze clinical characteristics and therapeutic efficacy of acute traumatic central cervical spinal-cord syndrome without fracture and dislocation,and to investigate its treatment methods. METHODS: Clinical data of 47 cases with acute traumatic central cervical spinal-cord syndrome without fracture and dislocation were retrospectively analyzed. There were 3 female and 44 male,with a mean age of 56.2 years (ranging from 46 to 73 years). According to Frankel classification, 30 cases were grade B, 14 cases were grade C and 3 cases were grade D. Forty-two cases were treated without operation, and 5 cases which had significant compression factors or segmental instability were treated with surgical treatment. Among them, 3 cases were treated through anterior approach and 2 cases were treated through posterior approach. Causes of injuries, mechanisms of injuries,findings of imaging, functional impairment of nerve,methods of treatment,and therapeutic efficacy were analyzed. RESULTS: The average followed-up time was 12 months (ranging form 3 to 84 months), 2 cases of non-operation were dead and 6 were loss. Three months after treatment, some complications occurrenced on 7 patients of non-operation. The majority bequeathed different degree impairment of intrinsic muscles of hands. CONCLUSION: Reduction and fixation through the pedicle of fractured vertebra is an effective therapeutic options to treat the thoracolumbar burst fractures and desearved clinical study and expansion. PMID- 19281028 TI - [Clinical study of transforaminal instrumented lumbar interbody fusion]. AB - OBJECTIVE: To study the effect and complications of transforaminal lumbar interbody fusion technique. METHODS: The medical records and radiographs of 40 patients undergone transforaminal lumbar interbody fusion between 2005 and 2007 were retrospectively reviewed. There were 49 segments with fusion. Preoperative and 1 year postoperative functional evaluation were graded with ODI and VAS scoring system. The height and angle of the intervertebral space and the fusion status were measured as well. RESULTS: All patients were followed-up for 12 to 24 months with the average of 18 mouths. There were no severity postoperative complications. The operation time averaged 160 min and average blood loss 510 ml. The effect results were excellent in 28 cases, good in 7 and fair in 5. The fusion rate was 100%. One year after operation, the pain relief in the VAS and the reduction of the ODI were significant (P < 0.01), the height and angle of the intervertebral space increased obviously (P < 0.05). Fifteen patients complained low back pain to some extent untill the last follow-up. CONCLUSION: Transforaminal lumbar interbody fusion can achieve satisfactory clinical and radiographic results especially for the failed back surgery syndrome. PMID- 19281029 TI - [Operative treatment of III degree injuries without fracture of ankle joint ligaments]. PMID- 19281030 TI - [Diagnosis and treatment for distal tibiofibular diastasis in the malleolus joint injuries]. PMID- 19281031 TI - [Treatment of trimalleolar fracture by fibular approach for exposing post malleolus]. PMID- 19281032 TI - [Clinical observation of treatment of talus fracture accompany dislocation emergency]. PMID- 19281033 TI - [Treatment of floating elbow injury with internal fixation of minimally invasive nailing]. PMID- 19281034 TI - [Clinical outcome of treatment for cervical disease through anterior cervical expansive decompression by the intervertebral space]. PMID- 19281035 TI - [Percutaneous kyphoplasty by sacculus expanding for treating vertebral compression fracture of osteoporosis]. PMID- 19281036 TI - [Thumb reversed neurocutaneous flap on dorsal-ulnar side to repair thumb soft tissue defects]. PMID- 19281037 TI - [Modified Ponseti method for the treatment of congenital clubfoot at early stage]. PMID- 19281038 TI - [Treatment of tibial avulsion fractures of the posterior cruciate ligament]. PMID- 19281039 TI - [Application of solis cage in anterior approach of cervical vertebrae]. PMID- 19281040 TI - [Effect of exogenous sodium hyaluronate on the level of MMP-9 in serum and joint synovia of patients with osteoarthritis]. PMID- 19281041 TI - [Study advancement of anatomy and biomechanics of posterior three type screw fixation techniques of axial]. AB - To explain the possibility of the placement of posterior three types screw instrument, and to compare the morphology and biomechanics difference among them. It is feasible and reliable for posterior three types screw instrument in anatomy and biomechanics. Axial pedicle screw would be best choice among posterior three types screw instrument. PMID- 19281042 TI - [Expression and regulation of matrix metalloproteases in osteoarthritic cartilage]. AB - The degradation of arthrodial cartilage is a typical characteristic in the pathogenesis of osteoarthritis. matrix metalloproteinases (MMPs) are the primary enzymes involved in extracellular matrix degradation of cartilage. The mechanism of MMPs in extracellular matrix degradation of cartilage is becoming clear with the in-depth study about MMPs, such as activation, activity regulation, related signal transduction pathways and transcription factors. This artice reviewed the activation, expression and regulation of MMPs in the related theory and empirical study of osteoarthritis cartilage. PMID- 19281043 TI - [Characteristics of human lipoaspirate-isolated mesenchymal stromal cells cultivated under a lower oxygen tension]. AB - In the present study we investigated the effect of low oxygen concentration in the cultivation medium on proliferation, viability and immunophenotype of human mesenchymal stromal cells isolated from lipoaspirate (lMSC). It was shown that proliferation activity of the cells under hypoxic conditions was, on average, 2.9 times higher compared to that under commonly accepted normoxic conditions. Redused oxygen level in the culture medium did not cause any change in IMSC viability and immunophenotype. Thus, permanent culture of lMSC in the medium with a lower oxygen tension can prove an efficient approach to obtain a higher mass of cells which maintain their characteristics in a shorter period of time, which can be of demand for regenerative medicine. PMID- 19281044 TI - [Comparison of fibroblasts-like cell differentiation capacities of human bone marrow, adipose tissue, hair papilla and dermal fibroblasts]. AB - We compared the morphology and differentiation characteristics of the human cells from bone marrow, adipose tissue, hair papilla and skin dermis. All cell types showed fibroblastic morphology. Immunofluorescent analysis showed that adipose tissue derived stem cells (ADAS) and hair papilla cells (HPC) expressed CD105, CD49d and STRO-1, bone marrow mesenchymal stem cells (BMSC) were absent for CD49, dermal fibroblasts (DFb) expressed CD49 and STRO-1 at low level. Populations of ADAS, BMSC and HPC had similar capacities to lipid and bone differentiation. Following exposure to appropriate induction stimuli, these cells changed phenotype and expressed specific cell markers. However, the rate and extent of HPC differentiation were lower in comparison with populations of ADAS and BMSC. We propose that all investigated cell populations contain primitive progenitor cells with mesenchymal stem cell properties. PMID- 19281045 TI - [Multiparametric study and cluster-analysis of the transformed cultures of human cells]. AB - The cells of nepatocarcinoma (HEp-G2), adenocarcinoma of large intestine (Caco 2), embryonal kidney (HEK-293), neuroblastoma (SH-SY5Y), rabdomyosarcoma (RD), and larynx cancer (Hep-2) were studied by the methods of scanning cytophotometry, cytochemistry and cytomorphometry during 96 h of cultivation. The density of monolayers, proliferation activity, the number of dead cells, DNA content in the nuclei and distribution of the cells in the population by this parameter, total DNA content in the nucleoli (circumnucleolar chromatin), the number of nucleoli in the nuclei, distribution of cells by their number, the volume and area of the nucleus surface, total volume and area of the nucleoli surface were determined. The data obtained were used in the treelike cluster analysis of the cultures by Pierson correlation. As a result, the SH-SY5Y culture was put in a separate cluster, while Caco-2, HEp-G2, HEK-293, Hep-2 and RD cultures were placed in the tree of another cluster. The least transformed culture of neuroblastoma (SH-SY5Y) had no relationship with other cultures, which showed various rate of similarity. The cultured HEK-293, Hep-2 and RD appeared to be close to each other by all parameters. The parameters studied are of different significance for the formation of general pattern of the cell cultures. The greatest "weight" is carried by the parameters, which characterize the population as whole: the density of the monolayer, mitotic coefficient and the number of dead cells. They are followed by the content of DNA in the nuclei, the total area of the nucleoli surface, and ratios of DNA content in the nucleoli to DNA content in the nucleus and of total surface of the nucleoli to the surface of the nuclei. Other parameters are not so significant. PMID- 19281046 TI - [The antigen of the differentiated basal disc cells of Hydra]. AB - A new antigenic marker of the differentiated basal disk cells of Hydra was characterized. An antigen named 3G11 was revealed by monoclonal antibody in granules of the basal disk gland cells of the ectoderm. The antigen appearance during budding, regeneration and ectopic foot formation evidences for the differentiation of the body column epithelial cells into basal disk gland cells. Antigen 3G11 is species-specific: among six hydra species investigated, the antigen was observed exclusively in polyps of vulgaris group which is a special taxon of the filum Hydra. Cell and tissue localization of the antigen 3G11 was similar to that of the well-established biochemical hydra marker, foot specific peroxidase, reported formerly. However, ELISA data suggest that the molecule bearing antigen 3G11 does not possess any peroxidase activity. Thus the new hydra antigenic marker 3G11 extends the number of previously used markers of differentiation and allows to improve the technique of the basal disk differentiated tissue identification. PMID- 19281047 TI - [Mechanisms of hepatocyte multinucleation in rats exposed to N nitrosodimethylamine]. AB - Mechanisms of hepatocyte multinucleation were investigated in rats exposed to N nitrosodimethylamine (NDMA).Using immunohistochemical reaction to y-tubulin it was established that the number of cells containing three and more centrosomes increased in 48 h after NDMA injection. It was shown that formation of extra centrosomes in hepatocytes was enhanced by oxidative stress induced by cytochromes P450 superfamily in the course of NDMA metabolism. NDMA administration led to a sharp increase in cytochrome P450 content in the liver, especially in 24 and 48 h (3.3 and 2.8 times respectively) after NDMA injection. Extensive staining of cytoplasm in the centrolobular hepatocytes was revealed by immunohistochemical reaction to cytochrome P450 2E1 in 24 and 48 h after the NDMA injection. Malone dialdehyde (the derivative of lipid peroxidation) was shown to increase 1.1-2.0 times, whereas catalase activity as of the antioxidative agent reduced to 1.1-1.3 times in that time. In 72-120 h after NDMA treatment, the number of cells with three or more centrosomes, the intensity of cytoplasmic staining, cytochrome P450 and malone dialdehyde contents in the liver were shown to decrease, whereas catalase activity increased. In 48 h after treatment, binucleated hepatocytes with various 3H-thymidine distribution in nuclei appeared in NDMA-treated cell populations evidencing of asynchronous DNA synthesis. Immunohistochemical reaction against Ki-67 proliferation marker revealed asynchronous nuclear proliferation activity in binucleated cells spreading not only to S-phase, but also to other phases of cell cycle, and namely G1, G2 and M. Thus, main mechanisms of hepatocyte multinucleation under NDMA exposure are accounted for hyperamplification of centrosomes as a consequence of oxidative stress and for asynchronous DNA synthesis in the nuclei of binucleate hepatocyte followed by asynchronous acytokinetic mitosis. PMID- 19281048 TI - [Influence of salicylic and succinic acids on the cytophysiological reactions of wheat infected by brown rust]. AB - The influence of the salicylic and succinic acids on the cytophysiological reactions of the plants and cellular structures of the fungus were investigated using the model of common wheat Triticum aestivum L. infected by brown rust. The experiments were performed on seedlings of the isogenic line of var. Thatcher with resistant gene Lr19. The salicylic and succinic acids accelerated and enhanced to a different extent the generation of active oxygen species and synthesis of callose and phenylpropanoids by the plant cells contacting with cellular structures of avirulent and virulent fungal clones. PMID- 19281049 TI - [Distribution of legume arabinogalactanprotein-extensin (AGPE) glycoproteins in symbiotically defective pea mutants with abnormal infection threads]. AB - The interface between the host cell and the microsymbiont is an important zone for development and differentiation during successive stages of the rhizobium legume symbiosis. Legume root nodule extensins, otherwise known as arabinogalactanprotein-extensin (AGPE) are abundant components of the infection thread matrix. We have characterized the origin and distribution of these glycoproteins at the symbiotic interface of root nodules of symbiotically defective mutants of pea (Pisum sativum L.) using immunogold localization with MAC265 an anti-AGPE monoclonal antibody. For mutants with defective growth of infection threads, the AGPE epitope was abundant in the extracellular matrix surrounding infected host cells in the central infected tissue of the nodule as well as being present in the lumen of Rhizobium-induced infection threads. This suggests a mis-targetting of AGPE as a consequence of abnormal growth of the infection threads. Furthermore, mutants in gene sym33 showed reduced labelling with MAC265, and in infection threads and droplets label was completely absent, a phenomenon not observed in wild-type nodules. This suggests an alteration in the composition of the infection thread matrix for sym33 mutants which may be correlated with the absence of endocytosis of rhizobia into the host cytoplasm. PMID- 19281050 TI - [Tyrosine hydroxylase in telencephalon and diencephalon of Rhodeus sericeus (Cyprinidae)]. AB - Immunohistochemical labeling of tyrosine hydroxylase was used to demonstrate catecholaminergic neuronal populations in the telencephalon and diencephalonof adult cypryniform fish Rhodeus sericeus. Various immunoreactive cell populations have been found in the telencephalon (ventral, central and lateral nuclei of ventral telencephalic area). Immunoreactive cells and fibers were discovered in dorsal nucleus of ventral telencephalic area and supracomissural nucleus in the caudal part of the telencephalon. In the diencephalon, periventricular nuclei (preoptic, periventricular nucleus of posterior tuberculum and periventricular organ) contained considerable TH-ergic cells. High activity of tyrosine hydroxylase was revealed in the pretectal, ventro-medial, ventro-lateral and suprachiasmatic nuclei. Periventricular hypothalamic nuclei also displayed high activity of tyrosine hydroxylase. Pseudounipolar neurons prevailed in all TH immunereactive structures of the telencephalon and diencephalon: numerous bipolar liquor-contacting cells were discovered in the periventricular nuclei. Large pear shaped cells and bipolar TH-ergic cells were found in posterior tuberculum. These cells may be functionally related to the dopamine-acquiring system. PMID- 19281051 TI - [Adaptive response in the different mitotic cycles after irradiation]. AB - The frequency of cells with chromosome aberrations and the number of aberrations per cell by metaphase analysis have been studied in the nonirradiated progeny of irradiated human blood lymphocytes. The DNA fragmentation (DNA double strand breaks) have simultaneously been investigated by the DNA comet assay. PHA stimulated lymphocytes have been irradiated in the adaptive dose 0.05 Gy 24 h and in the challenge dose 1 Gy 48 h after stimulation to study the adaptive response (AR). 5-bromodeoxyuridine have been added for the identification the first--the fourth mitoses. It has been discovered that the frequency of chromosome aberrations is increased is all mitotic cycles after challenge irradiation, the level of double strand breaks is increased too. The adaptive response in induced by the adaptive and challenge irradiation in the first and the second mitotic cycles (fixation 48 and 72 h after stimulation) for the most parts of individuals, but it is absent in the third and the fourth mitosis. Only chromatid aberrations are observed in the first mitosis, but chromosome aberrations--in the following mitosis. Investigation by the DNA comet assay have showed the adaptive response is noticed 48-72 h after stimulation but it is insignificant 96 h. The conclusion is that the genomic instability is observed in nonirradiated progeny irradiated lymphocytes; the adaptive response is manifested up to third mitosis and is explained by the decreasing of the number of the chromatid and chromosome aberrations and DNA fragmentation. We can suppose that double strand DNA breaks can be signaling damage for the adaptive response induction. PMID- 19281052 TI - [Change of mating type induced by actinomycin D in the ciliate Dileptus anser]. AB - Effect of actinomycin D on the expression and inheritance of mating types (MTs) in mature laboratory clones of the ciliate Dileptus anser was studied. Each mature clone of these ciliates isolated from natural population appears to belong to one of the three complementary MTs - MT I, MT II or MT III. Its MT does not vary in the course of further laboratory cultivation of the clone in a series of vegetative generations. However, after treatment with actinomycin D (15 microg/ml, 3 days), such clones became hereditarily unstable for their MTs. At weekly testing for MT over the course of at least 15 weeks after treatment (which corresponds to more than 100 cellular divisions), many subclones of the treated clone reversibly changed their MT for another, and (or) showed temporary state of immaturity or partial maturity. These data testify in favour of our hypothesis of epigenetic MT determination in D. anser. Considering that actinomycin D induces heritable destabilization of some characters in amebas Amoeba proteus which is obviously of epigenetic nature, this antibiotic can possibly be regarded as "epimutagen". PMID- 19281053 TI - Managing the risk of invasive breast cancer in women at risk for breast cancer and osteoporosis: the role of raloxifene. AB - Raloxifene hydrochloride is a selective estrogen receptor modulator (SERM) that has antiestrogenic effects on breast and endometrial tissue and estrogenic effects on bone, lipid metabolism, and blood clotting. Raloxifene significantly improves serum lipids and serum markers of cardiovascular disease risk, but it has no significant effect on the risk of primary coronary events. A meta-analysis of randomized, double-blind, placebo-controlled trials of raloxifene for osteoporosis showed the odds of fracture risk were 0.60 (95% confidence interval [CI] = 0.49-0.74) for raloxifene 60 mg/day compared with placebo. During 8 years of follow-up in an osteoporosis trial, the raloxifene group had a 76% reduction in the incidence of invasive ER-positive breast cancer compared with the placebo group. In the STAR trial, the incidence of invasive breast cancer was 4.30 per 1000 women-years with raloxifene and 4.41 per 1000 with tamoxifen; RR = 1.02; 95% CI, 0.82-1.28. The effect of raloxifene on invasive breast cancer was, therefore, equivalent to that of tamoxifen with more favorable rates of adverse effects including uterine malignancy and clotting events. Millions of postmenopausal women could derive net benefit from raloxifene through reduced rates of fracture and invasive breast cancer. PMID- 19281054 TI - Optimizing dosing frequencies for bisphosphonates in the management of postmenopausal osteoporosis: patient considerations. AB - Postmenopausal osteoporosis is common and underrecognized among elderly women. Osteoporotic fractures cause disability and disfigurement and threaten patients' mobility, independence, and survival. Care for incident fractures in this age group must go beyond orthopedic repair, to assessment and treatment of the underlying bone fragility. Fracture risk can be reduced by vitamin D and calcium supplementation along with antiresorptive drug treatment. First-line osteoporosis pharmacotherapy employs nitrogen-containing bisphosphonates. The inconvenience of daily oral treatment has motivated development of weekly, monthly, and intermittent oral regimens, as well as quarterly and yearly intravenous (i.v.) regimens. Ibandronate is the first bisphosphonate to have shown direct anti fracture efficacy with a non-daily regimen; it was approved for once-monthly oral dosing in 2005 and for quarterly i.v. dosing in 2006. Intermittent oral risedronate and yearly i.v. zoledronic acid were approved in 2007. Newly available regimens with extended dosing intervals reduce the inconvenience of bisphosphonate therapy and provide patients with a range of options from which to select a maximally sustainable course of treatment. This review discusses the development, efficacy, safety, and tolerability of extended-interval bisphosphonate regimens and examines their potential to improve patient acceptance and long-term success of osteoporosis treatment. PMID- 19281055 TI - Hyaluronic acid gel (Juvederm) preparations in the treatment of facial wrinkles and folds. AB - Soft tissue augmentation with temporary dermal fillers is a continuously growing field, supported by the ongoing development and advances in technology and biocompatibility of the products marketed. The longer lasting, less immunogenic and thus more convenient hyaluronic acid (HA) fillers are encompassing by far the biggest share of the temporary dermal filler market. Since the approval of the first HA filler, Restylane, there are at least 10 HA fillers that have been approved by the FDA. Not all of the approved HA fillers are available on the market, and many more are coming. The Juvederm product line (Allergan, Irvine, CA), consisting of Juvederm Plus and Juvederm Ultra Plus, was approved by the FDA in 2006. Juvederm is a bacterium-derived nonanimal stabilized HA. Juvederm Ultra and Ultra Plus are smooth, malleable gels with a homologous consistency that use a new technology called "Hylacross technology". They have a high concentration of cross-linked HAs, which accounts for its longevity. Juvederm Ultra Plus is used for volumizing and correcting deeper folds, whereas Juvederm Ultra is best for contouring and volumizing medium depth facial wrinkles and lip augmentation. Various studies have shown the superiority of the HA filler products compared with collagen fillers for duration, volume needed, and patient satisfaction. Restylane, Perlane, and Juvederm are currently the most popular dermal fillers used in the United States. PMID- 19281056 TI - Osteoporosis in men: its pathophysiology and the role of teriparatide in its treatment. AB - As the population ages, the burden of osteoporosis in men is expected to rise. Implementation of preventive measures such as falls prevention strategies, exercise and adequate calcium and vitamin D intake is recommended. However, when the diagnosis of osteoporosis is made, effective treatments need to be initiated to prevent fractures. As opposed to postmenopausal women, reduced bone formation is the predominant mechanism of age-related bone loss in men, making anabolic agents a logical treatment option for men with osteoporosis. Teriparatide is the only anabolic agent currently approved for treatment of osteoporosis in men. This paper summarizes the mechanism of action of teriparatide, as well as its tolerability and safety. Furthermore, the evidence supporting the efficacy of teriparatide treatment in men with osteoporosis is reviewed and its current role in the management of osteoporosis in men is discussed. PMID- 19281057 TI - Aromatase inhibitors, efficacy and metabolic risk in the treatment of postmenopausal women with early breast cancer. AB - The third-generation aromatase inhibitors (AIs), letrozole, anastrozole and exemestane, are becoming the first choice endocrine drugs for post-menopausal women with breast cancer, since they present greater efficacy when compared with tamoxifen in both adjuvant and metastatic setting. In particular, several large and well designed trials have suggested an important role for AIs in the adjuvant treatment of postmenopausal women with estrogen-receptor positive breast cancer either in the upfront, sequential or extended adjuvant mode. Overall, AIs are associated with a small but significant improvement in disease free survival. The expanding use of AIs in the treatment of early breast cancer means that individual patients will be exposed to the agents for longer durations, making it increasingly important to establish their long-term safety. This review focused on the effects of AIs on bone metabolism, serum lipids and cardiovascular risk. AIs have adverse effects on bone turnover with a reduction of bone mineral density and an increase in the rate of fragility fractures. With respect to tamoxifen AIs present lower thrombotic risk and a less favorable impact on lipid profile, whereas the true effects on cardiovascular risk still remain to be clarified. An adequate monitoring of bone mineral density (BMD) and lipid profile could be recommended for post-menopausal women candidate to AIs. PMID- 19281058 TI - Growth hormone and aging: a challenging controversy. AB - Although advanced age or symptoms of aging are not among approved indications for growth hormone (GH) therapy, recombinant human GH (rhGH) and various GH-related products are aggressively promoted as anti-aging therapies. Well-controlled studies of the effects of rhGH treatment in endocrinologically normal elderly subjects report some improvements in body composition and a number of undesirable side effects in sharp contrast to major benefits of GH therapy in patients with GH deficiency. Controversies surrounding the potential utility of GH in treatment of a geriatric patient are fueled by increasing evidence linking GH and cancer and by remarkably increased lifespan of GH-resistant and GH-deficient mice. Conservation of cellular signaling mechanisms that influence aging in organisms ranging from worms to mammals suggests that at least some of the results obtained in mutant mice are applicable to the human. We suggest that the normal, physiological functions of GH in promoting growth, sexual maturation and fecundity involve significant costs in terms of aging and life expectancy. Natural decline in GH levels during aging likely contributes to concomitant alterations in body composition and vigor but also may be offering important protection from cancer and other age-associated diseases. PMID- 19281059 TI - Backward disequilibrium in elderly subjects. AB - Backward disequilibrium is observed frequently in daily clinical practice. However, there are no epidemiological data concerning this postural disorder. Defined by a posterior position of the centre of mass with respect to the base of support, backward disequilibrium is abnormal postural behavior, usually characterized by a posterior trunk tilt in standing and sitting positions, which predisposes subjects to backward falls. Many afflictions whether they are somatic (degenerative, ischemic and traumatic brain lesions), psychosomatic (psychomotor disadaptation syndrome, confinement to bed, nonuse situations) or psychological (depression) can cause backward disequilibrium. A vicious circle of falls, and loss of autonomy can arise and this is the main consequence of backward disequilibrium. Thus, in this paper, we review backward disequilibrium in elderly subjects with regard to the causes, consequences, assessment, and management. PMID- 19281060 TI - Improvement of sensorimotor functions in old age by passive sensory stimulation. AB - Sensorimotor functions decrease in old age. The well-documented loss of tactile acuity in elderly is accompanied by deterioration ofhaptic performance and fine manipulative movements. Physical training and exercise can maintain sensorimotor fitness into high age. However, regular schedules of training require discipline and physical fitness. We here present an alternative interventional paradigm to enhance tactile, haptic, and fine motor performance based on passive, sensory stimulation by means of tactile coactivation. This approach is based on patterned, synchronous tactile stimulation applied to the fingertips for 3 hours. The stimulation drives plastic reorganizational changes in somatosensory cortex that affect perception and behavior: We demonstrate that following 3 hours of coactivation tactile acuity as well as haptic object exploration and fine motor performance are improved for at least 96 hours. Because this kind of intervention does not require active participation or attention of the subjects, we anticipate that coactivation is a prime candidate for future therapeutic interventions in patients with impaired sensorimotor abilities. It can be assumed that the maintenance and restoration of sensorimotor functions can ensure and preserve independence of daily living. Further optimizing of the stimulation protocol can be assumed to strengthen both the range and durability of its efficacy. PMID- 19281061 TI - Rare express saccades in elderly fallers. AB - OBJECTIVE: To examine horizontal saccades in elderly subjects with falling history; prior extensive screening was done to recruit subjects with falling history in the absence of pathology. METHODS: Twelve elderly with falling history were tested. Two testing conditions were used: the gap (fixation target extinguishes prior to target onset) and the overlap (fixation stays on after target onset) paradigms. Each condition was run at three viewing distances--20 cm, 40 cm, and 150 cm, corresponding to convergence angle at 17.1 degrees, 8.6 degrees, and 2.3 degrees, respectively. Eye movements were recorded with the photoelectric IRIS (Skalar medical). RESULTS: (i) like in healthy elderly subjects, elderly with falling history produce shorter latencies in the gap paradigm than in the overlap paradigm; (ii) their latencies are shorter at near distances (20 and 40 cm) relative to 150 cm for both paradigms; (iii) the novel result is that they fail to produce express latencies even in the conditions (near viewing distance and the gap task) known to promote high rates of express in adults (25%) or in healthy elderly (20%). Seven from the 10 healthy elderly produced express saccades at rates >12%, while 9 of the 12 older subjects with falling history showed no express saccades at all; the remaining 3 subjects showed low rates <12%. CONCLUSION: The quasi paucity of express saccades could be due to the disequilibrium of complex cortical/subcortical networks needed for making express saccades. The results support models suggesting specific network for express saccades; missing of such optomotor reflex may go along with missing other reflexes as well increasing the chances of falling. PMID- 19281062 TI - Correlation between myocardial enzyme serum levels and markers of inflammation with severity of coronary artery disease and Gensini score: a hospital-based, prospective study in Greek patients. AB - BACKGROUND: Our objective was to associate serum levels of myocardial enzymes and inflammatory biomarkers with severity of coronary artery disease (CAD). PATIENTS AND METHODS: 123 patients participated in our study, including 65 cases of acute myocardial infarction (MI), 27 cases of newly diagnosed CAD--without MI--and 31 controls. In all subjects, myocardial serum enzyme levels (creatine phosphokinase, aspartate aminotransferase, lactate dehydrogenase) and inflammatory indices (C-reactive protein, fibrinogen, white blood cells, and erythrocyte sedimentation rate) were measured. Patients were all submitted to coronary angiography and CAD severity was evaluated by Gensini score. RESULTS: Significant differences concerning enzyme serum levels and inflammatory indices were found to exist between the three study groups, being highest among patients with acute MI (p < 0.001). A significant association was demonstrated between Gensini score and serum enzyme levels as well as inflammatory biomarkers. CONCLUSIONS: Our findings suggest that serum levels of myocardial enzymes and inflammatory indices correlate with CAD severity in Greek patients. PMID- 19281063 TI - Food matrix and isoflavones bioavailability in early post menopausal women: a European clinical study. AB - The estrogenic effects of soy isoflavones (IF) on symptoms of menopause are of particular interest. The aim of the present study was to improve compliance of IF in two IF-enriched foods providing the same IF circulating levels in postmenopausal women. Forty-two healthy postmenopausal women (mean age: 53.28 years) were recruited for a randomized, crossover, multicenter trial conducted in the Netherlands, Italy and France. Over 18 days, volunteers were assigned to two groups and supplemented with two different IF-enriched foods (100 mg IF aglycones/two servings). The first group had to eat two biscuits daily for three days. After a wash-out period (11 d), they received cereal bars for three days. The second group started with the cereal bars and finished with biscuits. After IF intake, plasma and urinary levels of genistein, daidzein, O desmethyl angolensin and equol significantly increased and returned to baseline level after the washout period. There was no difference between biscuits and cereals bars intake, as shown by group values at each end of experimental period (day 4 or day 18). Both matrixes are comparable in terms of IF-circulating levels and could be used independently. PMID- 19281064 TI - Impact of cholinesterase inhibitors on behavioral and psychological symptoms of Alzheimer's disease: a meta-analysis. AB - OBJECTIVE: To determine the efficacy of cholinesterase inhibitors (ChEIs) in improving the behavioral and psychological symptoms of dementia (BPSD) in patients with Alzheimer's disease (AD). DATA SOURCES: We searched MEDLINE, Cochrane Registry, and the Cumulative Index to Nursing and Allied Health Literature (CINAHL) from 1966 to 2007. We limited our search to English Language, full text, published articles and human studies. DATA EXTRACTION: We included randomized, double-blind, placebo-controlled trials evaluating the efficacy of donepezil, rivastigmine, or galantamine in managing BPSD displayed by AD patients. Using the United States Preventive Services Task Force (USPSTF) guidelines, we critically appraised all studies and included only those with an attrition rate of less than 40%, concealed measurement of the outcomes, and intention to treat analysis of the collected data. All data were imputed into pre defined evidence based tables and were pooled using the Review Manager 4.2.1 software for data synthesis. RESULTS: We found 12 studies that met our inclusion criteria but only nine of them provided sufficient data for the meta-analysis. Among patients with mild to severe AD and in comparison to placebo, ChEIs as a class had a beneficial effects on reducing BPSD with a standard mean difference (SMD) of -0.10 (95% confidence interval [CI]; -0.18, -0.01) and a weighted mean difference (WMD) of-1.38 neuropsychiatry inventory point (95% CI; -2.30, -0.46). In studies with mild AD patients, the WMD was -1.92 (95% CI; -3.18, -0.66); and in studies with severe AD patients, the WMD was -0.06 (95% CI; -2.12, +0.57). CONCLUSION: Cholinesterase inhibitors lead to a statistical significant reduction in BPSD among patients with AD, yet the clinical relevance of this effect remains unclear. PMID- 19281065 TI - Purple urine bag syndrome in nursing homes: ten elderly case reports and a literature review. AB - Purple urine bag syndrome (PUBS) is a rare occurrence, in which the patient has a purple-colored urine bag following urinary catheterization for hours to days. Most of authors believe it is a mixture of indigo (blue) and indirubin (red) that becomes purple. Previous study showed that PUBS occurred predominantly in chronically catheterized, constipated women. We collected 10 elderly patients with PUBS in two nursing homes. The first two cases were identified by chart review in 1987 and 2003, and then later eight cases (42.1%) were collected among 19 urinary catheterized elderly in the period between January 2007 and June 2007. In the present report, PUBS probably can occur in any patients with the right elements, namely urinary tract infection (UTI) with bacteria possessing these enzymes, diet with enough tryptophan, and being catheterized. Associations with bed-bound state, Alzheimer's, or dementia from other causes are reflections of the state of such patients who are at higher risk for UTI, and hence PUBS occurred. Although we presented PUBS as a harmless problem, prevention and control of the nosocomial catheter-associated UTIs (CAUTIs) has become very important in the new patient-centered medical era. Thus, we should decrease the duration of catheterization, improve catheter care, and deploy technological advances designed for prevention, especially in the elderly cared for in nursing homes. PMID- 19281066 TI - Improvements in tongue strength and pressure-generation precision following a tongue-pressure training protocol in older individuals with dysphagia: three case reports. AB - Dysphagia, or difficulty swallowing, often occurs secondary to conditions such as stroke, head injury or progressive disease, many of which increase in frequency with advancing age. Sarcopenia, the gradual loss of muscle bulk and strength, can place older individuals at greater risk for dysphagia. Data are reported for three older participants in a pilot trial of a tongue-pressure training therapy. During the experimental therapy protocol, participants performed isometric strength exercises for the tongue as well as tongue pressure accuracy tasks. Biofeedback was provided using the Iowa Oral Performance Instrument (IOPI), an instrument that measures tongue pressure. Treatment outcome measures show increased isometric tongue strength, improved tongue pressure generation accuracy, improved bolus control on videofluoroscopy, and improved functional dietary intake by mouth. These preliminary results indicate that, for these three adults with dysphagia, tongue-pressure training was beneficial for improving both instrumental and functional aspects of swallowing. The experimental treatment protocol holds promise as a rehabilitative tool for various dysphagia populations. PMID- 19281067 TI - Chronomics, human time estimation, and aging. AB - BACKGROUND: Circadian rhythm stage affects many outcomes, including those of mental aging. METHODS: Estimations of 1 minute approximately 5 times/day for a year, 25 years apart, by a healthy male biomedical scientist (RBS), are analyzed by the extended cosinor. RESULTS: Cycles of a half-week, a week, approximately 30 days, a half-year and a year, in self-assessed 1-minute estimation by RBS between 25 and 60 years of age in health, are mapped for the first time, compared and opposite effects are found. For RBS at 60 vs at 25 years of age, it takes less time in the morning around 10:30 (P < 0.001), but not in the evening around 19:30 (P = 0.956), to estimate 1 minute. DISCUSSION: During the intervening decades, the time of estimating 1 minute differed greatly, dependent on circadian stage, being a linear decrease in the morning and increase in the evening, the latter modulated by a -33.6-year cycle. CONCLUSION: Circadian and infradian rhythm mapping is essential for a scrutiny of effects of aging. A approximately 30-day and a circannual component apparent at 25 years of age are not found later; cycles longer than a year are detected. Rhythm stages await tests as markers for timing therapy in disease. PMID- 19281068 TI - Health literacy in COPD. AB - If patients are to participate fully in their care and in the management of a long-term condition such as chronic obstructive pulmonary disease, good communication is essential. However, not all patients are able to use the written word and we need to be aware of the size of this problem and its implications for the way in which we give information and conduct medical consultations. The impact of health literacy on outcomes can be considerable and improvements can be made by being aware of the problem, offering information in several different forms, and by reinforcing the spoken word with pictorial images. PMID- 19281069 TI - Tissue regeneration as next-generation therapy for COPD--potential applications. AB - COPD is a major cause of chronic morbidity and mortality worldwide, and there is a need to develop more effective therapeutic strategies to replace specialized treatment such as lung transplantation. Recent studies suggest that recognition of apoptotic lung epithelial or endothelial cells may result in growth factors to stimulate cell replacement, and defects in these processes may contribute to the pathogenesis of COPD. Furthermore, recent animal and human studies have revealed that tissue-specific stem cells and bone marrow-derived cells contribute to lung tissue regeneration and protection, and thus administration of exogenous stem/progenitor cells or humoral factors responsible for activation of endogenous stem/progenitor cells may be a potent next-generation therapy for COPD. PMID- 19281070 TI - Use of azithromycin in the treatment of acute exacerbations of COPD. AB - Chronic bronchitis is a relatively common entity among patients with underlying chronic obstructive lung disease. Typical treatment includes pulmonary hygiene, bronchodilators, and antimicrobial therapy. In recent years, the duration of antimicrobial therapy in acute exacerbations of COPD has become shorter and shorter. This review summarizes the data on the use of the drug azithromycin for this particular patient population with a focus on 3-day and single-day therapy. PMID- 19281071 TI - Long-acting beta2-agonists (LABA) in chronic obstructive pulmonary disease: efficacy and safety. AB - COPD is characterized by progressive airflow obstruction which does not fully reverse to inhaled or oral pharmacotherapy. The management of patients with COPD has taken a totally new direction over the past 20 years, thank to the use of novel therapies aimed to improve and modify the natural history of COPD. Long acting bronchodilators, including long-acting beta2-agonists (LABAs), were introduced several years ago in order to enhance improvements in lung function, health status related quality of life, and reduce the rate of exacerbations. These effects can be boosted by the combination of LABAs with long-acting anticholinergic, and/or with inhaled corticosteroids. Inhaled LABAs are commonly well tolerated although adverse effects such as tremor and palpitations are occasionally troublesome. PMID- 19281072 TI - Role of T-lymphocytes and pro-inflammatory mediators in the pathogenesis of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the US and a major worldwide healthcare problem. The pathophysiologic mechanisms that drive development and progression of this disease are complex and only poorly understood. While tobacco smoking is the primary risk factor, other disease processes also appear to play a role. Components of the innate immune system (eg, macrophages and neutrophils) have long been believed to be important in the development of COPD. More recent evidence also suggests involvement of the adaptive immune system in pathogenesis of this disease. Here we will review the literature supporting the participation of T-cells in the development of COPD, and comment on the potential antigenic stimuli that may account for these responses. We will further explore the prospective contributions of T-cell derived mediators that could contribute to the inflammation, alveolar wall destruction, and small airway fibrosis of advanced COPD. A better understanding of these complex immune processes will lead to new insights that could result in improved preventative and/or treatment strategies. PMID- 19281074 TI - Treatment of mild chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is an epidemic in many parts of the world. Most patients with COPD demonstrate mild disease. The cornerstone of management of mild disease is smoking cessation, which is the only proven intervention to relieve symptoms, modify its natural history and reduce mortality. For asymptomatic patients, it is the only required therapy. Short acting bronchodilators can be added on an as needed basis for those with intermittent symptoms and regularly for those with persistent symptoms. Long acting bronchodilators can be substituted for those who remain symptomatic despite regular use of short-acting bronchodilators. Inhaled corticosteroids do not modify the natural history of COPD and as such cannot be recommended as standalone therapy for mild COPD. However, for patients with refractory and intractable symptoms, they may be used in combination with long-acting beta-2 agonists. Influenza and pneumococcal vaccination and pulmonary rehabilitation are other therapies that may be considered for select patients with mild disease. In this paper, we summarize the current standard of care for patients with mild COPD. PMID- 19281073 TI - ABCD of the phosphodiesterase family: interaction and differential activity in COPD. AB - Phosphodiesterases (PDEs) are important enzymes that hydrolyze the cyclic nucleotides adenosine 3'5'-cyclic monophosphate (cAMP) and guanosine 3'5'-cyclic monophosphate (cGMP) to their inactive 5' monophosphates. They are highly conserved across species and as well as their role in signal termination, they also have a vital role in intra-cellular localization of cyclic nucleotide signaling and integration of the cyclic nucleotide pathways with other signaling pathways. Because of their pivotal role in intracellular signaling, they are now of considerable interest as therapeutic targets in a wide variety diseases, including COPD where PDE inhibitors may have bronchodilator, anti-inflammatory and pulmonary vasodilator actions. This review examines the diversity and cellular localization of the isoforms of PDE, the known and speculative relevance of this to the treatment of COPD, and the range of PDE inhibitors in development together with a discussion of their possible role in treating COPD. PMID- 19281075 TI - Safety, tolerability and risk benefit analysis of tiotropium in COPD. AB - COPD is a chronic disease and, like many other chronic diseases, there is no treatment to reverse the severity of the disease except for lung transplant. To date, no inhaled medications have been shown to improve survival. Tiotropium bromide is a long-acting inhaled anticholinergic drug for the treatment of COPD that can improve lung function, reduce symptoms and exacerbations, and improve quality of life with once-daily dosing. It was initially approved and marketed in several countries in Europe in 2002 and then approved in the US in 2004. Tiotropium is generally well tolerated with dry mouth being the main adverse effect. Other adverse effects include constipation, tachycardia, blurred vision, urinary retention and increased intraocular pressure. Despite the recently raised concerns about an excess risk of cardiovascular adverse events with inhaled anticholinergic agents, the risk/benefit ratio of tiotropium appears still favorable given the favorable safety profile demonstrated in the UPLIFT study. However, caution should be advised in patients at high risk for cardiovascular disease given the paucity of data in such patients. PMID- 19281076 TI - Airway biomarkers of the oxidant burden in asthma and chronic obstructive pulmonary disease: current and future perspectives. AB - The pathogenesis of asthma and chronic obstructive pulmonary disease (COPD) has been claimed to be attributable to increased systemic and local oxidative stress. Detection of the oxidant burden and evaluation of their progression and phenotypes by oxidant biomarkers have proved challenging and difficult. A large number of asthmatics are cigarette smokers and smoke itself contains oxidants complicating further the use of oxidant biomarkers. One of the most widely used oxidant markers in asthma is exhaled nitric oxide (NO), which plays an important role in the pathogenesis of asthma and disease monitoring. Another oxidant marker that has been widely investigated in COPD is 8-isoprostane, but it is probably not capable of differentiating asthma from COPD, or even sensitive in the early assessment of these diseases. None of the current biomarkers have been shown to be better than exhaled NO in asthma. There is a need to identify new biomarkers for obstructive airway diseases, especially their differential diagnosis. A comprehensive evaluation of oxidant markers and their combinations will be presented in this review. In brief, it seems that additional analyses utilizing powerful tools such as genomics, metabolomics, lipidomics, and proteomics will be required to improve the specificity and sensitivity of the next generation of biomarkers. PMID- 19281077 TI - Treatment of respiratory failure in COPD. AB - Patients with advanced COPD and acute or chronic respiratory failure are at high risk for death. Beyond pharmacological treatment, supplemental oxygen and mechanical ventilation are major treatment options. This review describes the physiological concepts underlying respiratory failure and its therapy, as well as important treatment outcomes. The rationale for the controlled supply of oxygen in acute hypoxic respiratory failure is undisputed. There is also a clear survival benefit from long-term oxygen therapy in patients withchronic hypoxia, while in mild, nocturnal, or exercise-induced hypoxemia such long-term benefits appear questionable. Furthermore, much evidence supports the use of non-invasive positivepressure ventilation in acute hypercapnic respiratory failure. It application reduces intubation and mortality rates, and the duration of intensive care unit or hospital stays, particularly in the presence of mild to moderate respiratory acidosis. COPD with chronic hypercapnic respiratory failurebecame a major indication for domiciliary mechanical ventilation, based on pathophysiological reasoning and on data regarding symptoms and quality of life. Still, however, its relevance for long-term survival has to be substantiated in prospective controlled studies. Such studies might preferentially recruit patients with repeated hypercapnic decompensation or a high risk for death, while ensuring effective ventilation and the patients' adherence to therapy. PMID- 19281078 TI - Pathophysiological roles of microvascular alterations in pulmonary inflammatory diseases: possible implications of tumor necrosis factor-alpha and CXC chemokines. AB - Chronic obstructive pulmonary disease (COPD) and bronchial asthma are common respiratory diseases that are caused by chronic inflammation of the airways. Although these diseases are mediated by substantially distinct immunological reactions, especially in mild cases, they both show increased numbers of neutrophils, increased production of tumor necrosis factor-alpha (TNF-alpha) and poor responses to corticosteroids, particularly in patients with severe diseases. These immunological alterations may contribute strongly to airway structural changes, commonly referred to as airway remodeling. Microvascular alterations, a component of airway remodeling and caused by chronic inflammation, are observed and appear to be clinically involved in both diseases. It has been well established that vascular endothelial growth factor (VEGF) plays important roles in the airway microvascular alterations in mild and moderate cases of both diseases, but any role that VEGF might play in severe cases of these diseases remains unclear. Here, we review recent research findings, including our own data, and discuss the possibility that TNF-alpha and its associated CXC chemokines play roles in microvascular alterations that are even more crucial than those of VEGF in patients with severe COPD or asthma. PMID- 19281079 TI - Lung volume reduction surgery and lung transplantation in chronic obstructive pulmonary disease. AB - Medical treatment of emphysema does not alter the natural progression of the disease. Surgical techniques are an attractive conceptual approach to treat hyperinflation in these patients. Lung volume reduction surgery and lung transplantation are appropriate therapeutic options for a selected population with emphysema. We will review the available evidence to support these approaches. PMID- 19281081 TI - The role for S-carboxymethylcysteine (carbocisteine) in the management of chronic obstructive pulmonary disease. AB - Prescription of mucoactive drugs for chronic obstructive pulmonary disease (COPD) is increasing. This development in clinical practice arises, at least in part, from a growing understanding of the important role that exacerbation frequency, systemic inflammation and oxidative stress play in the pathogenesis of respiratory disease. S-carboxymethylcysteine (carbocisteine) is the most frequently prescribed mucoactive agent for long-term COPD use in the UK. In addition to its mucoregulatory activity, carbocisteine exhibits free-radical scavenging and anti-inflammatory properties. These characteristics have stimulated interest in the potential that this and other mucoactive drugs may offer for modification of the disease processes present in COPD. This article reviews the pharmacology, in vivo and in vitro properties, and clinical trial evidence for carbocisteine in the context of guidelines for its use and the current understanding of the pathogenic processes that underlie COPD. PMID- 19281080 TI - Skeletal muscle dysfunction in patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a debilitating disease characterized by inflammation-induced airflow limitation and parenchymal destruction. In addition to pulmonary manifestations, patients with COPD develop systemic problems, including skeletal muscle and other organ-specific dysfunctions, nutritional abnormalities, weight loss, and adverse psychological responses. Patients with COPD often complain of dyspnea on exertion, reduced exercise capacity, and develop a progressive decline in lung function with increasing age. These symptoms have been attributed to increases in the work of breathing and in impairments in gas exchange that result from airflow limitation and dynamic hyperinflation. However, there is mounting evidence to suggest that skeletal muscle dysfunction, independent of lung function, contributes significantly to reduced exercise capacity and poor quality of life in these patients. Limb and ventilatory skeletal muscle dysfunction in COPD patients has been attributed to a myriad of factors, including the presence of low grade systemic inflammatory processes, nutritional depletion, corticosteroid medications, chronic inactivity, age, hypoxemia, smoking, oxidative and nitrosative stresses, protein degradation and changes in vascular density. This review briefly summarizes the contribution of these factors to overall skeletal muscle dysfunction in patients with COPD, with particular attention paid to the latest advances in the field. PMID- 19281082 TI - Association of chronic obstructive pulmonary disease and obstructive sleep apnea consequences. AB - Obstructive sleep apnea syndrome (OSAS) and chronic obstructive pulmonary disease (COPD) are two diseases that often coexist within an individual. This coexistence is known as overlap syndrome and is the result of chance rather than a pathophysiological link. Although there are claims of a very high incidence of OSAS in COPD patients, recent studies report that it is similar to the general population. Overlap patients present sleep-disordered breathing associated to upper and lower airway obstruction and a reduction in respiratory drive. These patients present unique characteristics, which set them apart from either COPD or OSAS patients. COPD and OSAS are independent risk factors for cardiovascular events and their coexistence in overlap syndrome probably increases this risk. The mechanisms underlying cardiovascular risk are still unclear, but may involve systemic inflammation, endothelial dysfunction, and tonic elevation of sympathetic neural activity. The treatment of choice for overlap syndrome in stable patients is CPAP with supplemental oxygen for correction of upper airway obstructive episodes and hypoxemia during sleep. PMID- 19281083 TI - Diagnostic value of post-bronchodilator pulmonary function testing to distinguish between stable, moderate to severe COPD and asthma. AB - OBJECTIVE: The GOLD guidelines suggest that the presence of a post-bronchodilator forced expiratory volume in one second (FEV1) < 80% of the predicted value in combination with a FEV1/forced vital capacity (FVC) < 70% confirms the diagnosis of COPD. Limited data exist regarding the accuracy of these criteria to distinguish between COPD and asthma. The aim of this study therefore was to investigate the diagnostic value of post-bronchodilator lung function parameters in obstructive lung disease. METHODS: The pulmonary function tests of 43 (22 = COPD, 21 = asthma) patients with similar baseline characteristics were evaluated (baseline FEV were 55.7% +/- 7.6%, and 59.3% +/- 8.4% predicted for COPD and asthma, respectively). Bronchodilator responsiveness (BDR) was calculated according to three recognized pulmonary function test criteria. RESULTS: The first criteria, post-bronchodilator FEV1 < 80% of the predicted value in combination with a post-bronchodilator FEV1/FVC ratio of < 70%, had an accuracy of 70% to diagnose COPD. This combination was very sensitive (100%) in diagnosing COPD, but it was not specific (38%). The second BDR criteria, defined as an increase of < 12% and 200 mL of initial FEV1 and criterion number 3, an increase of < 9% of predicted FEV1, were less sensitive (55% and 59%, respectively), but more specific (81% and 76% respectively) to diagnose COPD. Our findings suggest that the current recommended spirometric indices are not optimal in differentiating between COPD and asthma. PMID- 19281084 TI - Determinants and predictors of the cost of COPD in primary care: a Spanish perspective. AB - OBJECTIVES: 1) To estimate the annual cost of patients with stable chronic obstructive pulmonary disease (COPD) followed in primary care in Spain; 2) To analyze the possible cost predictor variables. PATIENTS AND METHODS: A multicenter, epidemiological, observational, descriptive study. Sociodemographic data, severity of disease, associated comorbidity, treatment followed by patients, quality of life (SF-12 questionnaire), health care resource utilization in the previous 12 months and duration of working disability due to COPD were collected. RESULTS: A total of 10,711 patients (75.6% men; 24.4% women) with a mean age of 67.1 +/- 9.66 years were evaluated. The mean forced expiratory volume in one second (FEV1) value was 57.4 +/- 13.4%. The total cost per patient per year was Euro1,922.60 +/- 2,306.44. The largest component of this cost was hospitalization (Euro788.72 +/- 1,766.65), followed by cost of drugs (Euro492.87 +/- 412.15) and visits to emergency rooms (Euro134.32 +/- 195.44). Linear regression analysis found associated heart disease, FEV1, physical component of quality of life, number of medical visits (primary care physician, pneumologist and emergency room), hospital admissions (frequency and duration of stay) and duration of working disability to be significant predictors of the total annual cost. CONCLUSIONS: The total annual cost of a COPD patient followed in primary care in Spain was considered high in this study. The presence of associated heart disease, severity of airflow obstruction, physical component of quality of life, health care resource utilization and duration of work disability were found to be predictor of cost. PMID- 19281085 TI - Impact of social support and self-efficacy on functioning in depressed older adults with chronic obstructive pulmonary disease. AB - OBJECTIVE: The objective of this study was to examine the association between social support, self-efficacy, and functioning among a sample of depressed older adults with chronic obstructive pulmonary disease (COPD). METHODS: Participants were recruited immediately following admission to an acute pulmonary rehabilitation unit of a rehabilitation hospital. One hundred and fifty-six subjects completed assessments of depression, functioning, social support, and self-efficacy at admission to the rehabilitation unit. Regression analyses were conducted to evaluate the impact of different aspects of social support and self efficacy on overall functioning at admission. RESULTS: Controlling for depression, COPD severity, and age, subjective social support (p = 0.05) and self efficacy (p < 0.01) were associated with overall functioning. CONCLUSION: The perception of social support as well as self-efficacy are important constructs related to overall functioning among depressed older adults with COPD. Attention to these psychosocial variables in health management interventions may help maintain or improve the overall functioning of depressed COPD patients. PMID- 19281086 TI - Pulmonary and systemic hepatocyte and keratinocyte growth factors in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: The potential role of growth factors in chronic obstructive pulmonary disease (COPD) has begun to be addressed only recently and is still poorly understood. For this study, we investigated potential abnormalities of hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) in patients with COPD. METHODS: To this end, we compared the levels of HGF and KGF, measured by enzyme linked immunosorbent assay (ELISA), in bronchoalveolar lavage (BAL) fluid and in serum in 18 patients with COPD (62 +/- 9 yrs, forced expiratory volume in one second [FEV1] 57 +/- 12% ref, X +/- standard deviation of mean), 18 smokers with normal lung function (58 +/- 8 yrs, FEV1 90 +/- 6% ref) and 8 never smokers (67 +/- 9 yrs, 94 +/- 14% ref). RESULTS: We found that in BAL, HGF levels were higher in patients with COPD than in the other two groups whereas, in serum, HGF concentration was highest in smokers with normal lung function (p < 0.01). KGF levels were not significantly different between groups, neither in blood nor in BAL (most values were below the detection limit). CONCLUSIONS: These results highlight a different response of HGF in BAL and serum in smokers with and without COPD that may be relevant for tissue repair in COPD. PMID- 19281087 TI - Mental disorders and quality of life in COPD patients and their spouses. AB - In the current study, the prevalence of the most common psychological disorders in COPD patients and their spouses was assessed cross-sectionally. The influence of COPD patients' and their spouses' psychopathology on patient health-related quality of life was also examined. The following measurements were employed: Forced expiratory volume in 1 second expressed in percentage predicted (FEV1%), Shuttle-Walking-Test (SWT), International Diagnostic Checklists for ICD-10 (IDCL), questionnaires on generic and disease-specific health-related quality of life (St. George's Respiratory Questionnaire (SGRQ), European Quality of Life Questionnaire (EuroQol), a modified version of a Disability-Index (CDI)), and a screening questionnaire for a broad range of psychological problems and symptoms of psychopathology (Symptom-Checklist-90-R (SCL-90-R)). One hundred and forty three stable COPD outpatients with a severity grade between 2 and 4 (according to the GOLD criteria) as well as 105 spouses took part in the study. The prevalence of anxiety and depression diagnoses was increased both in COPD patients and their spouses. In contrast, substance-related disorders were explicitly more frequent in COPD patients. Multiple linear regression analyses indicated that depression (SCL-90-R), walking distance (SWT), somatization (SCL-90-R), male gender, FEV1%, and heart disease were independent predictors of COPD patients' health-related quality of life. After including anxiousness of the spouses in the regression, medical variables (FEV1% and heart disease) no longer explained disability, thus highlighting the relevance of spouses' well-being. The results underline the importance of depression and anxiousness for health-related quality of life in COPD patients and their spouses. Of special interest is the fact that the relation between emotional distress and quality of life is interactive within a couple. PMID- 19281088 TI - Noninvasive positive pressure ventilation in acute respiratory failure due to COPD vs other causes: effectiveness and predictors of failure in a respiratory ICU in North India. AB - OBJECTIVES: To determine the effectiveness of noninvasive positive pressure ventilation (NIPPV), and the factors predicting failure of NIPPV in acute respiratory failure (ARF) due to chronic obstructive pulmonary disease (COPD) versus other causes of ARF. PATIENTS AND METHODS: This was a prospective observational study and all patients with ARF requiring NIPPV over a one-and-a half year period were enrolled in the study. We recorded the etiology of ARF and prospectively collected the data for heart rate, respiratory rate, arterial blood gases (pH, partial pressure of oxygen in the arterial blood [PaO2], partial pressure of carbon dioxide in arterial blood [PaCO2]) at baseline, one and four hours. The patients were further classified into two groups based on the etiology of ARF as COPD-ARF and ARF due to other causes. The primary outcome was the need for endotracheal intubation during the intensive care unit (ICU) stay. RESULTS: During the study period, 248 patients were admitted in the ICU and of these 63 (25.4%; 24, COPD-ARF, 39, ARF due to other causes; 40 male and 23 female patients; mean [standard deviation] age of 45.7 [16.6] years) patients were initiated on NIPPV. Patients with ARF secondary to COPD were older, had higher APACHE II scores, lower respiratory rates, lower pH and higher PaCO2 levels compared to other causes of ARF. After one hour there was a significant decrease in respiratory rate and heart rate and decline in PaCO2 levels with increase in pH and PaO2 levels in patients successfully managed with NIPPV. However, there was no difference in improvement of clinical and blood gas parameters between the two groups except the rate of decline of pH at one and four hours and PaCO2 at one hour which was significantly faster in the COPD group. NIPPV failures were significantly higher in ARF due to other causes (15/39) than in ARF-COPD (3/24) (p = 0.03). The mean ICU and hospital stay and the hospital mortality were similar in the two groups. In the multivariate logistic regression model (after adjusting for gender, APACHE II scores and improvement in respiratory rate, pH, PaO2 and PaCO2 at one hour) only the etiology of ARF, ie, ARF-COPD, was associated with a decreased risk of NIPPV failure (odds ratio 0.23; 95% confidence interval, 0.58-0.9). CONCLUSIONS: NIPPV is more effective in preventing endotracheal intubation in ARF due to COPD than other causes, and the etiology of ARF is an important predictor of NIPPV failure. PMID- 19281089 TI - C-reactive protein correlates with tissue oxygen availability in patients with stable COPD. AB - BACKGROUND: Arterial oxygen tension, oxygen delivery to tissue, and systemic inflammation are recognized as pivotal factors in the progression of chronic obstructive pulmonary disease (COPD). However, interconnections between systemic inflammation and tissue oxygen availability are scantly investigated. Tissue oxygen availability depends on arterial PaO2, oxygen concentration, hemoglobin oxygen affinity (P50), and hemoglobin oxygen binding capacity (ceHb). As the integrated changes of those indices are summarized by oxygen extraction tension (PaO2x), the objective of this study was to explore the association between C reactive protein (CRP) blood levels and either PaO2x or each of its determinants, in stable COPD. MATERIALS AND METHODS: Blood CRP and oxygen status of arterial blood were measured at rest while breathing room air in 44 moderate to severe stable COPD patients. PaO2x was calculated along the shape of oxygen binding curve as the oxygen tension resulting from removal of 2.3 mmol of oxygen per liter of blood. Multiple linear regression analysis was performed with PaO2, ceHb, and P50 as independent variables, and CRP as the dependent variable, adjusting for age and sex. The analysis was repeated using PaO2x as a sole independent variable. RESULTS: Multiple linear regression analysis indicated that ceHb, PaO2, and P50, were significant and independent predictors of CRP (R2 = 0.52, p < 0.0001). PaO2x alone was an even stronger predictor of CRP (R2 = 0.62, p < 0.0001). CONCLUSIONS: These findings indicate that physiological determinants of tissue oxygen availability are independently associated with CRP blood levels. Thus, improvement of tissue oxygen availability is a central therapeutic option to modulate the severity of systemic inflammatory processes in patients with COPD. PMID- 19281090 TI - Caregivers for people with end-stage lung disease: characteristics and unmet needs in the whole population. AB - INTRODUCTION: End-stage lung disease (ESLD) (predominantly caused by chronic obstructive pulmonary disease and restrictive lung disease) is a significant cause of death. Little is known about community care for people with ESLD especially in the period leading to death. This paper describes demographic characteristics of caregivers, and key characteristics of the deceased irrespective of specialist service utilization. METHODS: The South Australian Health Omnibus is an annual, random, face-to-face, cross-sectional survey conducted statewide. For the last eight years questions about end of life have been asked of 3000 respondents annually (participation rate 77.9%). Directly standardized to the whole population, this study describes people who cared for someone with ESLD until death. RESULTS: One third (6370/18267) had someone die in the last five years from a terminal illness, 644 from ESLD (3.5% of respondents; 10.2% of deaths). One in five (20.8%) provided physical care: 43 respondents provided day-to-day and 63 provided intermittent hands-on care for an average of 40.1 months (SD 56.9). Caregivers were on average 51.2 years old (range 17-85; SD 16.5) and one in five was a spouse. Additional support to provide physical care was an unmet need by 17% of caregivers. The deceased were an average of 73.9 years old (range 47-92; SD 10.4). Only 31.1% were assessed as 'comfortable' or 'very comfortable' in the last fortnight of life. DISCUSSION: Given the health consequences of caregiving, caregivers of people with ESLD would benefit from prospectively defining their needs given the time for which intense caregiving is provided. PMID- 19281091 TI - Higher lung deposition with Respimat Soft Mist inhaler than HFA-MDI in COPD patients with poor technique. AB - Aerosols delivered by Respimat Soft Mist Inhaler (SMI) are slower-moving and longer-lasting than those from pressurized metered-dose inhalers (pMDIs), improving the efficiency of pulmonary drug delivery to patients. In this four-way cross-over study, adults with chronic obstructive pulmonary disease (COPD) and with poor pMDI technique received radiolabelled Berodual (fenoterol hydrobromide 50 microg/ipratropium bromide 20 microg) via Respimat SMI or hydrofluoroalkane (HFA)-MDI (randomized order) on test days 1 and 2, with no inhaler technique training. The procedure was repeated on test days 3 and 4 after training. Deposition was measured by gamma scintigraphy. All 13 patients entered (9 males, mean age 62 years; FEV1 46% of predicted) inhaled too fast at screening (peak inspiratory flow rate [IF]: 69-161 L/min). Whole lung deposition was higher with Respimat SMI than with pMDI for untrained (37% of delivered dose vs 21% of metered dose) and trained patients (53% of delivered vs 21% of metered dose) (P(Sign-Test) = 0.15; P(ANOVA) < 0.05). Training also improved inhalation profiles (slower average and peak IF as well as longer breath-hold time). Drug delivery to the lungs with Respimat SMI is more efficient than with pMDI, even with poor inhaler technique. Teaching patients to hold their breath as well as to inhale slowly and deeply increased further lung deposition using Respimat SMI. PMID- 19281092 TI - Tiotropium and exercise training in COPD patients: effects on dyspnea and exercise tolerance. AB - BACKGROUND: Exercise training improves exercise tolerance in chronic obstructive pulmonary disease (COPD). Tiotropium 18 microg once daily induces sustained bronchodilation throughout the day and reduces hyperinflation, one of the pathophysiological factors contributing to exertional dyspnea in COPD patients. AIM: To determine whether tiotropium enhances the effects of exercise training in patients with COPD. DESIGN: Multicenter, 25 week randomized, double-blind, placebo-controlled, parallel-group study. SETTING: Twelve Italian Pulmonary Units practicing pulmonary rehabilitation. PATIENTS AND INTERVENTION: Two hundred thirty four COPD patients (196 males; mean age: 67.4 +/- 7.6; forced expiratory volume at 1 second (FEV1): 41.4 +/- 13.0% predicted) were randomised to tiotropium 18 microg or placebo inhalation capsules taken once daily. Both groups underwent a 8 week pulmonary rehabilitation program (PR) consisting of 3 exercise training session per week. MEASUREMENTS: Baseline, at the end of PR and after 12 weeks, patients completed pulmonary function testing, six minute walking test (6MWT), the Baseline and Transition Dyspnea Index (BDI and TDI), and the St. George's Respiratory Questionnaire (SGRQ). RESULTS: Relative to placebo, tiotropium had larger trough and post-study drug FEV1 responses on all test days. At the end of and 12 weeks following PR, patients on tiotropium showed no statistically significant differences in 6MWT compared to patients on placebo. Compared to the period immediately prior to PR, the mean improvement in 6MWT was only 29.7 meters (7.1%) for the combined cohort. Mean TDI focal scores at the end of PR were 3.60 for tiotropium and 2.25 for placebo (p < 0.01). At 12 weeks after PR, TDI focal scores were 2.71 for tiotropium and 2.11 for placebo (p = 0.16). Reduction in all four SGRQ component scores, indicating an improvement in health related quality of life, was observed for the tiotropium group over the duration of the study compared to placebo but the differences were not statistically significant. During the study period, there were fewer exacerbations and exacerbation days in the tiotropium group. CONCLUSION: Although significant improvements were observed with perceived dyspnea, compared to placebo, the addition of tiotropium to pulmonary rehabilitation did not improve the 6MWT. PMID- 19281093 TI - Suppressive activity of tiotropium bromide on matrix metalloproteinase production from lung fibroblasts in vitro. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is characterized by airway remodeling with an accumulation of inflammatory cells. There is also increasing evidence that metalloproteinases (MMPs) may contribute to the pathogenesis of COPD, but the influence of agents that used for the treatment of COPD is not well understood. OBJECTIVE: We evaluated whether tiotropium bromide hydrate (TBH), a M3 muscarinic receptor antagonist, could inhibit MMP production from lung fibroblasts (LFs) in response to tumor necrosis factor (TNF)-alpha stimulation. METHODS: LFs were established from normal lung tissues taken from patients with lung tumors. LFs (5 x 10(5) cells/ml) were stimulated with TNF alpha in the presence of various concentrations of TBH. After 24 h, culture supernatants were obtained and assayed for the levels of MMPs and tissue inhibitor of metalloproteinases (TIMPs) by ELISA. The influence of TBH on mRNA expression of MMPs and TIMPs in 4h-cultured cells was also examined by real-time RT-PCR. Furthermore, nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) in LFs treated with TBH for 4h was examined by ELISA. RESULTS: TBH at more than 15 pg/ml inhibited the production of MMP-2 from LFs after TNF-alpha stimulation, whereas TIMP-1 and TIMP-2 production was scarcely affected by TBH through the suppression of both mRNA expression and transcription factor, NF kappaB, activation in LFs induced by TNF-alpha stimulation. CONCLUSION: These results suggest that the attenuating effect of TBH on MMP-2 production from LFs induced by inflammatory stimulation may be additional beneficial therapeutic effects not directly relating to its bronchodilatory effects. PMID- 19281094 TI - Self efficacy measurement and goal attainment after pulmonary rehabilitation. AB - BACKGROUND: Little is known about self-efficacy in COPD and its role as a mediator of goal attainment after rehabilitation. We hypothesized that higher baseline self efficacy may facilitate goal attainment. METHODS: 48 COPD patients completed pulmonary rehabilitation, self-efficacy was measured using the COPD Self Efficacy Scale (CSES). Personal goals were used as self reported outcome. Relationships were evaluated between CSES and St George's Hospital Respiratory Questionnaire (SGRQ); depression; using Brief Assessment Schedule Cards (BASDEC), London Chest Activity of Daily Living Scale (LCADL) and exercise tolerance; using Six-Minute Walking Distance (6MWD) and muscle strength. RESULTS: 74 Stable COPD patients, mean FEV1 1.2 (0.6) 1, age 68.1 (10.2) years were recruited. 51 patients completed rehabilitation and 48 of those CSES and reported goal attainment. 94 goals were documented. Baseline self efficacy did not differ according to whether goal was achieved or not. Relationships were evident between CSES and SGRQ (r = -0.53), 6MWD (r = 0.36), BASDEC (r = -0.31), LCADL (r = -0.33) (all p < or = 0.01), but not FEV1, pack years or muscle strength. There was a significant improvement in CSES scores pre to post rehabilitation, mean difference (95% CI) 0.27 (0.04 to 0.51). CONCLUSIONS: Self efficacy, using the CSES, improves with rehabilitation but baseline self efficacy does not appear to influence goal attainment. PMID- 19281095 TI - Effect of noninvasive mechanical ventilation in elderly patients with hypercapnic acute-on-chronic respiratory failure and a do-not-intubate order. AB - Noninvasive mechanical ventilation (NIMV) is effective in the treatment of patients with acute respiratory failure (ARF). It proved to reduce the need of endotracheal intubation (ETI), the incidence of ETI-associated pneumonia, and mortality compared to nonventilated patients. A particular aspect concerns the outcome of NIMV in patients referring to an emergency room (ER) for ARF, and with a do-not-intubate (DNI) status due to advanced age or critical conditions. The aim of our study is to assess the outcome of NIMV in a group of elderly patients with acute hypercapnic ARF who had a DNI status. An overall number of 62 subjects (30 males, 32 females, mean age 81 +/- 4.8 years, range 79-91 years) referred to our semi-intensive respiratory department were enrolled in the study. The underlying diseases were severe chronic obstructive pulmonary disease (COPD) in 50/62 subjects, restrictive thoracic disorders in 7/62 subjects, and multiorgan failure in 5/62 subjects. Fifty-four/62 patients were successfully treated with NIMV while 2/62 did not respond to NIMV and were therefore submitted to ETI (one survived). Among NIMV-treated patients, death occurred in 6 patients after a mean of 9.9 days; the overall rate of NIMV failure was 12.9%. Negative prognostic factors for NIMV response proved to be: an older age, a low Glasgow Coma Score, a high APACHE score at admission, a high PaCO2 after 12 hours and a low pH both after 1 and 12 hours of NIMV. We conclude that elderly patients with acute hypercapnic ARF with a DNI status can be successfully treated by NIMV. PMID- 19281096 TI - Geographic differences in clinical characteristics and management of COPD: the EPOCA study. AB - AIMS: Data on differences in clinical characteristics and management of COPD in different countries and settings are limited. We aimed to characterize the profile of patients with COPD in a number of countries and their treatment in order to evaluate adherence to recommendations of international guidelines. METHOD: This was an observational, international, cross-sectional study on patients with physician-diagnosed COPD. Demographic and clinical characteristics, risk factors, and treatment were collected by their physician via an internet web based questionnaire developed for the study. RESULTS: A total of 77 investigators from 17 countries provided data on 833 patients. The countries with the highest number of patients included were: Argentina (128), Ecuador (134), Spain (162), and Hong Kong (153). Overall, 79.3% were men and 81% former smokers, with a mean FEV1 = 42.7%, ranging from 34.3% in Hong Kong to 58.8% in Ecuador. Patients reported a mean of 1.6 exacerbations the previous year, with this frequency being significantly and negatively correlated with FEV1 (%) (r = -0.256; p < 0.0001). Treatment with short-acting bronchodilators and theophyllines was more frequent in Ecuador and Hong Kong compared with Spain and Argentina, and in patients belonging to lower socioeconomic levels (p < 0.0001 for all comparisons). Inadequacy of treatment with inhaled corticosteroids and theophyllines was high, with significant differences among countries. CONCLUSIONS: Differences in the clinical characteristics and management of COPD were significant across countries. Adherence to international guidelines appears to be low. Efforts should be made to disseminate and adapt guidelines to the socioeconomic reality of different settings. PMID- 19281097 TI - 'Long time coming': are we on the brink of universal health care coverage? PMID- 19281098 TI - From hospital to nursing facility: factors influencing decisions. AB - This study addresses the factors influencing decisions to send medicine-surgical (med-surg) patients home or to nursing facilities (NFs). The sample (n = 7,852) was taken from a large, urban, teaching, med-surg unit where discharges were documented and data collected over a two-and-a-half-year period. Using logistical regression, the factors found to most influence the decision were age (z = 26.99, p = .000; odds = 1.06); patients diagnosed with "musculoskeletal system" problems (z = 11.07, p = .000; odds = 5.36); and needing skilled professional care (z = 15.03, p = .000; odds = .21) or nonprofessional personal care (z = 6.62,p = .000; odds = 2.32). Having less effect, but important information for discharge planners, was being an African American (z = 3.82, p = .000; odds = .76) or Latino (z = -3.96, p = .000; odds = .54). A review of the literature found limited knowledge of the factors that influence hospital patients, family members, and professionals, including social workers, to make the decision to recommend home care or NF care. PMID- 19281099 TI - Education: a complex and empowering social work intervention at the end of life. AB - Education is a frequently used social work intervention. Yet it seems to be an underappreciated and a deceptively complex intervention that social workers may not be adequately prepared to use. Reliable, accessible information is essential as it helps prevent unnecessary crises, facilitates coping, and promotes self determination. This article conceptualizes education as a fundamental social work intervention and discusses the role social workers play in providing information that is both empowering and culturally sensitive. In particular, this article focuses on social workers working with patients and families facing life threatening situations, including those in hospice and other end-of-life care settings. After reviewing the relevant literature and theory and exploring the inherent complexities of educational interventions, the authors recommend strategies for more effectively helping patients and families access the information they need. PMID- 19281100 TI - The influence of community-based services on the burden of spouses caring for their partners with dementia. AB - Despite the vast literature on caregiver stress, few studies have explored how community services affect the stress process for spousal caregivers. The current study explores the differential effects of emotional and tangible support provided by family and friends and by formal services, and caregivers' perceptions of community services on spousal caregivers' burden. The study used a survey design with a sample of 85 spousal caregivers caring for their partners with dementia in the community. The study found that in-home services, as they are currently offered, do little to reduce the burden of spouses caring for their partners with dementia. Rather the most effective service is the provision of adult day programs, which provide not only respite for the spousal caregiver, but also opportunities for social interaction for their partners with dementia. This study further found that spousal caregivers experience a relatively high level of service-related stress. However, when examined alongside care recipient behavioral challenges and frequency of day program use, caregivers' perceptions of and experiences with the service system did not uniquely explain their burden. Implications of the findings for policy, research, and practice are discussed. PMID- 19281101 TI - Roles, responsibilities, and relationships among older husbands caring for wives with progressive dementia and other chronic conditions. AB - Men are playing greater roles in the provision of care for older adults with chronic health conditions. Husbands, in particular, encounter many role transformations as they witness their wives grow in levels of dependence as a result of their illnesses. This qualitative study examines the changes that occurred in the roles, responsibilities, and relationships of 17 husbands who were providing care for their wives with memory loss and other chronic health conditions. The results suggest that husbands experience changes in the ways that they adapt their marital roles to the new roles they assume as caregivers. It was found that the husbands had to learn how to form new types of relationships with their ailing wives. Implications for geriatric social workers and other health care providers are discussed. PMID- 19281102 TI - The changing face of opioid addiction: prescription pain pill dependence and treatment. PMID- 19281103 TI - Medicare Prescription Drug, Improvement, and Modernization Act of 2003: implications for the future of health care. PMID- 19281104 TI - Building capacity for evidence we can believe in: the argument for social change as an evidence-based practice agenda. PMID- 19281105 TI - The global increase in dental caries. A pending public health crisis. AB - A current review of the available epidemiological data from many countries clearly indicates that there is a marked increase in the prevalence of dental caries. This global increase in dental caries prevalence affects children as well as adults, primary as well as permanent teeth, and coronal as well as root surfaces. This increase in dental caries signals a pending public health crisis. Although there are differences of opinion regarding the cause of this global dental caries increase, the remedy is well known: a return to the public health strategies that were so successful in the past, a renewed campaign for water fluoridation, topical fluoride application, the use of fluoride rinses, a return to school oral health educational programs, an emphasis on proper tooth brushing with a fluoride dentifrice, as well as flossing, a proper diet and regular dental office visits. If these remedies are not initiated, there could be a serious negative impact upon the future oral health (and systemic health) of the global community, as well as a strain on the dental profession along with a major increase in the cost of dental services. PMID- 19281106 TI - Fracture strength of endodontically treated molars transfixed horizontally by a fiber glass post. AB - PURPOSE: To assess the effect of a horizontally transfixed fiber glass post placed between buccal and palatal surfaces, on the fracture strength of endodontically treated molar teeth with MOD cavities, either restored with resin based composite, or not. METHODS: 75 sound maxillary human third molars were extracted, embedded in acrylic resin blocks and randomly assigned to five groups (n=15). Group A (sound teeth), (control) and Groups B, C, D and E, which were subjected to the following procedures after endodontic treatment: GB--(MOD+Endo), GC--(MOD+Endo+Post), GD--MOD and composite restoration (MOD+Endo+CR), GE- (MOD+Endo+Post+CR). The specimens were stored in distilled water at 37 degrees C for 24 hours. Later, a compressive force was applied by means of a universal testing machine at 1 mm/minute speed, parallel to the long axis of the teeth until fracture occurred. RESULTS: The means of the results (N) followed by the same letter represent no statistical difference by ANOVA and Tukey (P<0.05): GA = 4289.8 (+/- 1128.9)a, GB = 549.6 (+/- 120.7)b, GC = 1474.8 (+/- 338.1)c, GD = 1224.7 (+/- 236.0)c, GE = 2645.4 (+/- 675.1)d. In the analysis of qualitative variables, there was a tendency to cusp fracture in all groups except for Group C. The fiber glass post transfixed horizontally in a MOD cavity significantly increased the fracture resistance of the teeth restored with resin composite. PMID- 19281107 TI - Efficacy of calcium hydroxide, Er:YAG laser or gaseous ozone against Enterococcus faecalis in root canals. AB - PURPOSE: To evaluate the efficacy of Ca(OH)2, Er:YAG laser or gaseous ozone (either alone or combined with instrumentation and various irrigants) against Enterococcus faecalis in root canals. METHODS: 180 extracted, human, single rooted teeth were divided into four groups of 45 teeth each. In Group 1 root canal enlargement up to ISO-size 60 (MAF) was performed, whereas only initial shaping (MAF ISO-size 40) was carried out in Groups 2 to 4. After sterilization all teeth were inoculated with E. faecalis and incubated for 3 days, followed by evaluation of CFU. Subsequently, root canal enlargement up to ISO-size 60 was performed in Groups 2 to 4 using NaCl solution (0.9%) in Group 2, NaOCl (1%) in Group 3 and CHX (0.2%) in Group 4. Finally, each group of 45 teeth was subdivided into three groups (n= 15 each) either applying Ca(OH)2 for 7 days, using Er:YAG laser radiation for 30 seconds or gaseous ozone for 120 seconds, followed by final evaluation of CFU. RESULTS: Both in Groups 1 and 2 the median reduction of bacteria after application of Ca(OH)2 (factor 10(4) each) and ozone (in Group 1: factor 5 x 10(3); in Group 2: factor 5 x 10(4)), respectively, was significantly higher than after Er:YAG laser treatment (factor 102 each, Mann-Whitney test). The antibacterial efficacy was significantly increased by the additional use of NaOCl or CHX as irrigants in all subgroups (Groups 3 and 4) compared to corresponding subgroups of Group 1 (Mann-Whitney test). PMID- 19281108 TI - Spectrophotometric evaluation of color match to VITA classical shade guide of four different veneering porcelain systems for metal ceramic restorations. AB - PURPOSE: To determine by a spectrophotometric analysis the variations in color between the fabricated shade of four different porcelain systems and the intended shade when applied in a set thickness. METHODS: Four porcelain systems (Duceram Kiss, VITA Omega, Wieland Reflex, Ivoclar IPS d.SIGN) for metal ceramic restorations were selected. Three disk-shaped (15 mm diameter, 0.3 mm thickness) specimens per group were made for three different shades of the A color scale of the Vitapan classical shade guide. On a Cr-Co alloy, 0.15 mm of porcelain opaque and 1.0 mm of translucent porcelain was applied. The porcelain stratification was executed according to the manufacturers' indications. The color measures were made by the clinical spectrophotometer Easyshade. RESULTS: No statistically significant differences were found among the ceramic systems examined. The recorded differences in color between A2 porcelain disks and corresponding VITA shade tabs were closer (mean delta E = 2.50) than those recorded for shade A3 (mean delta E = 3.84) and shade A3.5 (mean delta E= 3.94) (P<0.05). PMID- 19281109 TI - Whitening effect and morphological evaluation of hydroxyapatite materials. AB - PURPOSE: To measure the efficacy of the whitening effect of non-oxidizing and non acidic nano- and micro-hydroxyapatite materials on the enamel surface and to evaluate the surface changes after treatment. METHODS: Three hydroxyapatite suspensions and two hydroxyapatite mixtures in dissolvable polymer films were applied to a total of 30 extracted caries-free human premolars. After the last material application, a hydrodynamic shear force was generated and applied for 2 minutes to all teeth to simulate mechanical loading of the surface. The tooth color was measured with a dental spectrophotometer. The mean changes of the L*a*b* values between different measurements in each group were expressed as delta E and were analyzed with ANOVA and the Tukey's post-hoc test. RESULTS: The groups of the nano-hydroxyapatite, the hydroxyapatite-nanocrystals and the tricalcium-phosphate exhibited significant delta E values between baseline and after hydrodynamic shear force application (P<0.05). In conclusion, the materials used in the study are very promising alternatives to oxidizing bleaching agents. PMID- 19281110 TI - Effects of acidic primer/adhesives on primary and permanent dentin. AB - PURPOSE: To evaluate the quality of primary and permanent dentin by Fourier transformed Raman spectroscopy (FT-Raman), and scanning electron microscopy/energy-dispersive spectroscopy (SEM/EDS). METHODS: The middle dentin of crowns was reached by carbide bur abrading providing a uniform smear layer. Self-etching primers were applied in order to simulate the etching of self etching adhesive systems. The groups were (n = 6): G1 (primary dentin smear layer); G2 (primary dentin etched by primer of Clearfil Protect Bond); G3 (primary dentin etched by Adper Prompt); G4 (permanent dentin smear layer); G5 (permanent dentin etched by primer of Clearfil Protect Bond); G6 (permanent dentin etched by Adper Prompt). SEM/EDS were made in order to obtain additional elemental data to complement FT-Raman. FT-Raman data were submitted to cluster analysis. RESULTS: Overall, FT-Raman showed differences between primary and permanent dentin concerning organic content, but not for inorganic content. FT Raman showed differences in the organic content between primary and permanent dentin after self-etching primer use. HEMA usage caused molecular changes in the organic content, while phosphoric acidic ester caused molecular changes in the inorganic content of primary and permanent dentin. The SEM/EDS identified C, O, P, and Ca, which could not replace ions to change mineral molecular arrangement. Both organic and mineral content arrangements were similar after self-etching primers action. The organic content of dentin was modified by both Clearfil PB primer and Adper Prompt for primary dentin but only by Clearfil PB for permanent dentin. The inorganic content of primary middle dentin was similar to the inorganic content of permanent middle dentin, even when Adper Prompt is used. PMID- 19281111 TI - Effect of acid etching time on the degradation of resin-dentin bonds in primary teeth. AB - PURPOSE: To investigate the influence of etching time on the degradation of resin dentin bonds produced in primary teeth. METHODS: 40 primary molars were randomly divided into four groups according to the adhesive system, Single Bond (SB) and Clearfil SE Bond (CSEB), and acid etching time. SB was applied to dentin after phosphoric acid etching for 15 or 7 seconds, whereas CSEB was applied after the application of SE Primer for 20 or 10 seconds. Resin composite crowns were built up followed by the production of specimens with a cross-sectional area of 0.49 mm2, which were further divided according to the storage condition, 24 hours, 6 and 12 months in water. Data were submitted to ANOVA and Tukey tests (alpha=0.05). RESULTS: After 24 hours there was no significant difference between bond strengths produced by the adhesive systems, irrespective of the acid etching time. Water storage for 6 and 12 months significantly reduced bond strengths of SB, especially when the dentin was acid etched for 15 seconds. For CSEB, no significant alteration in bond strength was seen up to the storage period of 12 months for both etching times. PMID- 19281112 TI - Influence of curing mode intensities on cell culture cytotoxicity/genotoxicity. AB - PURPOSE: This study determined the cytotoxic/genotoxic effect of different curing modes on cell culture. METHODS: A thin layer of lymphocyte cultures was cured applying three different curing modes of Bluephase C8 LED curing unit. Cultures were exposed to light directly or through a layer (2 mm) of polymerized resin composite sample. Cells were analyzed using trypan blue exclusion test, acridine orange/ethidium bromide dyeing technique, and alkaline comet assay. RESULTS: Only low intensity mode after direct exposure significantly increased the number of nonviable lymphocytes detected using trypan blue. All curing procedures significantly increased the number of apoptotic lymphocytes regardless whether the exposure occurred directly or through the composite. Low intensity mode in direct exposure significantly elevated DNA migration compared to other curing modes. 1 hour after exposure significant increase in tail length and intensity for all modes and procedures was detected. However, DNA damage measured for cultures cured by low intensity mode was higher compared to the other two modes; thus, despite of curing light intensity, longer curing time leads to greater cytotoxicity/genotoxicity in cell culture. PMID- 19281113 TI - Association between staining by caries detector dye and the corresponding mineral density in dentin caries. AB - PURPOSE: To examine an association between coloration (red, pink) resulting from staining with Caries Detector Dye (CDD) and the corresponding mineral density in dentin caries lesions determined by transverse microradiography (TMR). METHODS: CDD coloration of the lesion sections (approx. 190 microm) prepared from extracted caries teeth was photographed, and the corresponding relative mineral densities (RMD: relative values to the sound dentin) were obtained by TMR. A parallel study was performed using artificially demineralized and then remineralized dentin lesions. RESULTS: The mean RMD values in the naturally black pigmented, red- and pink-stained portions were 46 +/- 26.7%, 64 +/- 24.5%, and 80 +/- 15.1%, respectively. There were statistical differences in the RMD values among the three portions, as well as a wider range of RMD value distributions in the red and black-pigmented portions than in the pink portion.Even among the black-pigmented and red portions, much higher RMD values more than 90% were observed in several lesions, which were close to the mineral density of the sound (unaffected) dentin tissue. On the other hand, the remineralized surface layer of artificially demineralized lesions did not show the red coloration, and there seemed a threshold value of mineral density (approx. 21%), beyond which the red coloration was not observed. Similar threshold value was noted in the remineralized lesion body. This study showed a remarkable discrepancy regarding the RMD value for the red staining behavior between the naturally occurring caries and artificial carious lesion. PMID- 19281114 TI - IPS Empress inlays luted with a self-adhesive resin cement after 1 year. AB - PURPOSE: To prospectively compare the clinical performance of two different resin composites for luting IPS Empress inlays and onlays. METHODS: 83 IPS Empress restorations were placed in 30 subjects. All restorations were inserted under rubber dam. 43 inlays/onlays were luted with a self-adhesive resin cement [RelyX Unicem (RX)]. A multistep adhesive (Syntac) was used with Variolink II low viscosity (SV) and served as control (n=40). The restorations were evaluated after 2 weeks: Baseline = 1st recall (R1), after 6 months (R2) and after 1 year (R3) by two calibrated examiners using the modified USPHS criteria. RESULTS: From R1 to R3, one failure was noticed in the SV group (R2) due to marginal enamel chipping. After 1 year of clinical service, SV revealed significantly better results regarding color match and integrity inlay (Mann-Whitney U-test, P< 0.05). No statistically significant differences were observed between SV and RX for the remaining criteria (Mann-Whitney U-test, P>0.05). PMID- 19281115 TI - Effect of nanofillers' size on surface properties after toothbrush abrasion. AB - PURPOSE: To investigate the effect of filler-particle size of experimental and commercial resin composites, undergoing toothbrush abrasion, on three surface properties: surface roughness (SR), surface gloss (G) and color stability (CS). METHODS: Four model (Ivoclar/Vivadent) and one commercial resin composite (Tokuyama) with varying filler-size from 100-1000 nm were examined. Six discs (10 mm x 2 mm) from each product were prepared and mechanically polished. The samples were then submitted to 20,000 brushing strokes in a toothbrush abrasion machine. SR parameters (Ra, Rt and RSm), G, and CS were measured before and after toothbrush abrasion. Changes in SR and G were analyzed by 2-way ANOVA, with Bonferroni post hoc test. CS values were submitted to one-way ANOVA and Bonferroni post hoc test (alpha=0.05). RESULTS: Initial G values ranged between 73-87 gloss units (GU) and were reduced after toothbrush abrasion to a range of 8 64 GU. Toothbrush abrasion resulted in significant modifications in SR and G amongst the materials tested, attributed to filler sizes. There was statistically significant difference in color (delta E* ranged from 0.38-0.88). Filler size did not affect color stability. Toothbrush abrasion resulted in rougher and matte surfaces for all materials tested. Although the individual differences in surface roughness among filler sizes were not always significant, the correlation showed a trend that larger filler sizes resulted in higher surface roughness after abrasion for the SR parameters Ra and Rt (r = 0.95; r = 0.93, respectively). RSm showed an increase after toothbrush abrasion for all resin composites, however no significant correlation was detected (r = 0.21).There was a significant correlation between G and Ra ratios (r = - 0.95). PMID- 19281116 TI - NPSG compliance problems hospitals face now; where do you stack up? PMID- 19281117 TI - Study: safety checklist reduces mortality. PMID- 19281118 TI - Accountability, vigilance make this hospital grade A. PMID- 19281119 TI - How MN's reporting laws have changed the state. PMID- 19281120 TI - [Anatomic and histological characteristics of vagina reconstructed by McIndoe method]. AB - BACKGROUND/AIM: Congenital absence of vagina is known from ancient times of Greek. According to the literature data, incidence is 1/4 000 to 1/20 000. Treatment of this anomaly includes non-operative and operative procedures. McIndoe procedure uses split skin graft by Thiersch. The aim of this study was to establish anatomic and histological characteristics of vagina reconstructed by McIndoe method in Mayer Kuster-Rockitansky Hauser (MKRH) syndrome and compare them with normal vagina. METHODS: The study included 21 patients of 18 and more years with congenital anomaly known as aplasio vaginae within the Mayer Kuster Rockitansky Hauser syndrome. The patients were operated on by the plastic surgeon using the McIndoe method. The study was a retrospective review of the data from the history of the disease, objective and gynecological examination and cytological analysis of native preparations of vaginal stain (Papanicolau). Comparatively, 21 females of 18 and more years with normal vaginas were also studed. All the subjects were divided into the groups R (reconstructed) and C (control) and the subgroups according to age up to 30 years (1 R, 1C), from 30 to 50 (2R, 2C), and over 50 (3R, 3C). Statistical data processing was performed by using the Student's t-test and Mann-Writney U-test. A value of p < 0.05 was considered statistically significant. RESULTS: The results show that there are differences in the depth and the wideness of reconstructed vagina, but the obtained values are still in the range of normal ones. Cytological differences between a reconstructed and the normal vagina were found. CONCLUSION: A reconstructed vagina is smaller than the normal one regarding depth and width, but within the range of normal values. A split skin graft used in the reconstruction, keeps its own cytological, i.e. histological and, so, biological characteristics. PMID- 19281121 TI - [Quantitative analysis of lamellar bodies in amniotic fluid as fetal pulmonary maturity indicator]. AB - BACKGROUND/AIM: Although lamellar bodies have been the center of interest over the last years, the published results of fetal pulmonary maturity determination according to their concentration in amniotic fluid are controversial. The aim of this study was to determine the significance of lamellar bodies, as well as the ratio lecithin/sphingomyelin (L/S) in amniotic fluid for the assessment of fetal pulmonary maturity. METHODS: This prospective 2-year study included 102 female examinees, ranging from 17 to 44 years of age, in whom lamellar bodies concentrations in amniotic fluid were determined to check the efficacy of the applied therapy for obtaining arteficial fetal pulmonary maturity. The shake test was applied as a comparative test for determining a quantitative L/S ratio. To determine a fetus maturity and development stage we followed up biparietal diameter, abdominal circumference, femure length, ponderal index at birth and body mass. RESULTS: Out of a total of 102 amniocenteses within a period from 26th to 40th gestation week only 70 results were considered due to 32 unknown neonatal outcomes. Biparietal diameter was 224-362 mm, femur length 56 - 78 mm, ponderal index 1.22-2.84, fetus body mass 1300-4 350 g. There was found a significant relation between gestation age and lamellar bodies concentration (R = 0.396398, p < 0.01), as well as between gestation age and the ratio L/S (R = 0.691297, p < 0.01). Also, there was a significant correlation of lamellar bodies concentration to the ratio L/S determined (R = 0.493609, p < 0.01). CONCLUSION: Determination of lamellar bodies concentration values is a reliable method to confirm fetal pulmonary maturity. PMID- 19281122 TI - Our first experiences in applying an original method for removal of ABO isoagglutinins in ABO-incompatible kidney recipients. AB - BACKGROUND/AIM: Due to improved methods for removal of ABO isoagglutinins and novel immunosuppressive protocols, short and long-term outcome in blood group incompatible is similar to blood group compatible kidney transplantation. The aim of this study was to determine the efficacy of our original method for removal of ABO isoagglutinins from the blood in ABO-incompatible kidney allograft recipients. METHOD: Between 2006 and 2008 twelve patients were transplanted from ABO incompatible living donors. Titers of ABO isoagglutinins were 4-128 (IgG). Immunosuppressive therapy started 14 days before kidney transplantation with rituximab, followed by a triple therapy (prednisone + tacrolimus + mycophenolate mofetil) and the first plasma exchange (PE) procedure, in which one plasma volume was substituted with albumin and saline on day 7 before transplantation. For selective extracorporeal immunoadsorption, the removed plasma was mixed with donor blood type filtered red blood cells, centrifuged and the supernatant separated and preserved. In the next PE procedure, the removed plasma was replaced with immunoadsorbed plasma, and so on. Titers of ABO agglutinins, renal allograft function and survival were followed-up. RESULTS: The pre-transplant treatment consisting of 1-5 PE procedures and immunosuppressive therapy resulted in target ABO agglutinins titers below 4. During a 10-24 month follow-up three patients had an early acute rejection, one patient acute rejection and hemolytic anemia, two patients surgical complications and one of them lost his graft. In the post-transplant period, the titers of ABO antibodies remained below 4. All the patients had stable kidney allograft function with mean serum creatinine +/- SD of 129 +/- 45 micromol/l at the end of the study. CONCLUSION: Our method for removal of ABO antibodies was effective in a limited series of patients and short term follow-up. PMID- 19281123 TI - [Clinical efficacy of ampicillin in treatment of acute odontogenic abscess]. AB - BACKGROUND/AIM: Antibiotics choice and the duration of their application in the therapy of acute odontogenic abscess is considered to be controversial. The aim of this study was to investigate the clinical efficacy of ampicillin in treatment of acute odontogenic abscess and to assess the antimicrobial susceptibility of the isolated bacteria in early phase of abscess development. METHODS: This study included 60 patients with acute odontogenic abscess who were surgically treated (extraction of teeth and/or abscess incision) divided into two groups, ampicillin group and surgical group (without antibiotic treatment). RESULTS: In the ampicillin group of patients treatment lasted on the average 4.67 days, while in the surgical group 6.17 days. A total of 78 bacterial strains were isolated from 60 patients. The most often bacteria were found to be Gram-positive facultative anaerobs (68/78). The most common bacteria isolated were Viridans streptococci (43/78). Susceptibility of isolated bacteria to ampicillin were 70.5%. CONCLUSION: Peroral use of ampicillin, after surgicel treatment in an early phase of dentoalveolar abscess development, statistically significantly reduced the time of clinical symptoms of acute odontogenic abscess in comparison to surgical treatment only. The isolated bacterial strains in an early phase of dentoalveolar abscess development showed a high sensitivity to ampicillin. PMID- 19281124 TI - Do stone size and impaction influence therapeutic approach to proximal ureteral stones? AB - BACKGROUND/AIM: Primary therapeutic approach to lumbar ureteral stones is still contraversial. The aim of the study was to investigate the influence of stone impaction and size on the effectiveness of proximal ureteral stone lithotripsy. METHODS: A total of 123 patients with proximal ureteral stones were investigated in this prospective study performed in a 10-month period. The patients were divided into the group I--86 patients treated with extracorporeal shock wave lithotripsy (ESWL) and the group II--37 patients treated with "Swiss" Lithoclast. In the group I, 49 stones (57%) were classified as impacted, while 20 stones (23.3%) were larger than 100 mm2. In the group II, 26 stones (70.3%) were impacted, and 11 stones (29.7%) were larger than 100 mm2. Stones were defined as impacted by the radiographic, echosonographic as well as endoscopic findings in the group II of patients. Stone size was presented in mm2. Chemical composition of stones were almost the same in both groups of the patients. RESULTS: Generally, there was no statistically significant difference in the treatment success between the groups. However, stones larger than 100 mm2 were statistically more successfully treated endoscopically, while there was no statistical difference in the treatment success of impacted stones between these two groups. CONCLUSION: ESWL can by considered as primary first therapeutic approach in treatment of all proximal ureteral stones except for stones larger than 100 mm2 that should primarily be treated endoscopically. PMID- 19281125 TI - [Implication of late diagnosis for survival of patients with colorectal carcinoma]. AB - BACKGROUND/AIM: Colorectal cancer (CRC) is one of the most frequent diseases and early diagnosis has a potential role to improve survival. The aim of this study was to analyze influence of delay in diagnosis on survival in patiens with colorectal cancer. METHODS: A total of 119 patients with pathohystological diagnosis of CRC were included in the study. They were operated at our Department for Surgery from 2000 to 2002. They were divided into two groups according to the duration of symptoms: early operated patients - EOP (symptoms were presented for 3 months) and late operated patients - LOP (duration of symptoms was more than 3 months). Follow-up period was 5 year. RESULTS: Weight loss, intermittent abdominal pain and anorexia were more frequent in LOP (p < 0.01). Young age, blood in stool, and tumor localized in rectum were dominant characteristics in EOP (p < 0.05). Overall delay in diagnosis was 2.19 +/- 0.79 months in EOP and 11.37 +/- 5.68 months in LOP. There was highly statistically significant difference between these two groups (p < 0.01). Overall survival was 44.75%. Five years survival was 65.9% in the group of EOP and 26.5% in the group of LOP (chi2 = 28.16, p < 0.01) Weight loss was dominant characteristics in the patients who did not survive five years (chi2 = 14.26, p < 0.01). A period of 2 months in delay in diagnosis is "cut-off' value in prediction of death (sensitivity of 75.5% and specificity of 90.3%). CONCLUSION: A delay in diagnosis and stage of the disease are highly significant factors of patients with CRC survival. In everyday medical practice higher importance should be put on weight loss, intermittent abdominal pain, change in bowel habits, as well as on syderopenic anaemia. PMID- 19281126 TI - [Pharmacodynamic and pharmacokinetic effects of flumazenil and theophylline application in rats acutely intoxicated by diazepam]. AB - BACKGROUND/AIM: The majority of symptoms and signs of acute diazepam poisoning are the consequence of its sedative effect on the CNS affecting selectively poli synaptic routes by stimulating inhibitory action of GABA. The aim of the present study was to examine the effects of combined application of theophylline and flumazenil on sedation and impaired motor function activity in acute diazepam poisoning in rats. METHODS: Male Wistar rats were divided in four main groups and treated as follows: group I--with increasing doses of diazepam in order to produce the highest level of sedation and motor activity impairment; group II- diazepam + different doses of flumazenil; group III--diazepam + different doses of theophylline; group IV--diazepam + combined application of theophylline and flumazenil. Concentrations of diazepam and its metabolites were measured with LC MS. The experiment was performed on a commercial apparatus for spontaneous motor activity registration (LKB-Farad, Sweden). Assessment of diazepam-induced neurotoxic effects and effects after theophylline and flumazenil application was performed with rotarod test on a commercial apparatus (Automatic treadmill for rats, Ugo Basile, Italy). RESULTS: Diazepam in doses of 10 mg/kg and 15 mg/kg produced long-time and reproducible pharmacodynamic effects. Single application of flumazenil or theophylline antagonized effects of diazepam, but not completely. Combined application of flumazenile and theophylline resulted in best effects on diazepam-induced impairment of motoric activity and sedation. As a result of theopylline application there was better elimination of diazepam and its metabolites. CONCLUSION: Combined application of flumazenil and theophylline resulted in the best antidotal effects in the treatment of diazepam poisoned rats. These effects are a result of different mechanisms of their action, longer half-life of theophylline in relation to that of flumezenil and presumably the diuretic effect of theophylline. PMID- 19281127 TI - [K-ras mutation predictive significance in platinum based chemotherapeutic protocols in patients with advanced non-small cell lung cancer]. AB - BACKGROUND/AIM: K-ras oncogene is mutated in about 20% of lung cancer. The purpose of this study was to investigate the predictive significance for therapeutic response of K-ras mutations in advanced non-small cell lung cancer (NSCLC) patients. METHODS: Bronchial aspirate samples were assessed prior to platinum-based chemotherapy start in 39 patients with stage IIIb or IV NSCLC. K ras mutations at codons 12 and 13 were analyzed by single strand conformation polymorphisam (SSCP) and allele specific oligonucleozide hybridisation of polymerase chain reaction (PCR) of the patient's DNA present in bronchial aspirate. After two cycles of chemotherapy the patients were subjected to response evaluation. RESULTS: Of 39 patients 10 (25.5%) demonstrated K-ras mutations, while 29 (74.4%) patients had not. There were no significant differences between these two groups of patients with respect to baseline patient caracteristics. Partial response to the therapy had 16 (41%), no changes 14 (36%), and progressive disease 9 (23%) patients. There was a tendency to higher response rate for patients without K-ras mutations versus those with mutations, but not statistically significant (p = 0.14). CONCLUSION: There was no significant predictive value for therapeutic response of K-ras mutations for advanced non-small cell lung cancer. PMID- 19281128 TI - Assessment of quality of care in acute postoperative pain management. AB - BACKGROUND/AIM: Managing of acute postoperative pain should be of great interest for all hospital institutions, as one of the key components of patients satisfaction, which indicates quality, as well as the outcome of treatment. The aim of this study was to assess the quality of nursing care in managing acute postoperative pain and to establish factors which influence patients assessment of the same. METHOD: The investigation was conducted on the sample of 135 patients hospitalized in surgical clinics of the Clinical Centre of Vojvodina in Novi Sad in the form of cross-sectional study, by interviewing patients during the second postoperative day and collecting sociodemographic variables, type of surgical procedure and applied analgesic therapy which were taken from their medical documentation. The modified questionnaire of the Strategic and Clinical Quality Indicators in Postoperative Pain Management (SCQIPP) was used as the instrument of the investigation. The data were processed with suitable mathematical statistics methods such as multivariate analyses of variance (MANOVA), discriminative and other parametric procedures and methods. Roy's test, Pearson's coefficient contingency (X), multiple correlation coefficient (R) were conducted amongst other univariant procedures. RESULTS: The mean score for the individual items of SCQIPP questionnaire was between 2.0 and 4.7 (scale range 1 5) and the percentage of patients answers "strongly agree" ranged from 4.4 to 77%. The smallest number of positive answers were given by the patients for the item "In order to assess pain intensity, some of the staff asked me at least once in the morning, in the afternoon and in the evening to show the number from 0 10". Most of the patients (57%) evaluated severe pain during the previous 24 hours, as moderate pain, which represents significantly greater number of patients which complain of severe pain and mild pain (p < 0.001). The analysis of patients evaluation (MANOVA p < 0.05 and discriminative p < 0.05) indicates the existence of significant difference between the assessment of nursing care quality in managing acute postoperative pain as regards to clinics as the place for pain management, patient's sex and his expectations. Evaluation from "communication" subscale gives the greatest contribution (24.9%) to the difference between the observed clinics, and the greatest contribution (25.7%) to the difference in evaluation of nursing care quality as regards to patients' sex has the evaluations from "procedure" subscale. CONCLUSION: The results of this study show a useful evidence and identify aspects of nursing care in postoperative management of acute pain which are still to be improved. According to the patients' answers the priority should be given to a regular assessment of the intensity of postoperative pain and evaluation of the effects of analgesic therapy. PMID- 19281129 TI - [Multiple gigantic renal cysts]. AB - INTRODUCTION: Cystic renal lesions are very heterogeneous lesions which differ in ethiopathogenesis, morphological and clinical manifestations, and also in evolution and therapy. Classification of cystic lesions is complex, symptomatology is poor, and diagnosis is based on complete radiological diagnostic procedures. CASE REPORT: We presented a 20-year old patient with mild subjective symptoms. Objectively, he was without positive clinical signs and changes in biochemistry of blood. Using ultrasonography (US) multiple serous simple cysts were found in both kidneys. Using computed tomography (CT) multiple serous cysts were found, without changes in cystic walls, with preserved renal parenchyma and without cystic changes on other parenchymatous organs. CONCLUSION: Although renal cystic lesions are frequent in adult population, this is a rare example of a young adult man with simple, gigantic, serous cysts which do not produce clinical manifestations nor functional renal difficulty so far. PMID- 19281130 TI - [Anterior pituitary lobe atrophy as late complication of hemorrhagic fever with renal syndrome]. AB - INTRODUCTION: Hemorrhagic fever with renal syndrome (HFRS) is acute infective multisystemic disease followed by febrility, hemorrhages and acute renal insufficiency. Bleeding in the anterior pituitary lobe leading to tissue necrosis occurs in acute stage of severe clinical forms of HFRS, while atrophy of the anterior pituitary lobe with diminution of the gland function occurs after recovery stage. CASE REPORT: We presented a patient with the development of chronic renal insufficiency and hypopituitarism as complication that had been diagnosed six years after Hantavirus infection. Magnetic resonance of the pituitary gland revealed atrophy and empty sella turcica. CONCLUSION: Regarding frequency of this viral infection and its endemic character in some parts of our country partial and/or complete loss of pituitary function should be considered during the late stage of HFRS. PMID- 19281131 TI - [Two centuries of urologic endoscopy]. PMID- 19281132 TI - Difference in phototoxicity of cyclodextrin complexed fullerene [(gamma CyD)2/C60] and its aggregated derivatives toward human lens epithelial cells. AB - The water-soluble fullerene derivative gamma-cyclodextrin bicapped C(60) [(gamma CyD)(2)/C(60), CDF0] has several clinical applications, including use as a drug carrier to bypass the blood ocular barriers or a photosensitizer to treat tumors in photodynamic therapy. We have assessed the potential ocular toxicity of (gamma CyD)(2)/C(60) and its aggregated derivatives induced by UVA and visible light in vitro in human lens epithelial cells (HLE B-3). Cell viability using the MTS assay demonstrated that 2 microM (gamma-CyD)(2)/C(60) was highly phototoxic to HLE B-3 cells with UVA irradiation, while no effect was observed in the presence of visible light or when maintained in the dark. In contrast, the aggregated derivative (CDF150) showed neither cytotoxicity nor any phototoxic effect even at 30 microM with either UVA or visible light irradiation. In lens cells treated with (gamma-CyD)(2)/C(60), phototoxicity was manifested as apoptosis. Singlet oxygen production measurement using the EPR/TEMP trapping technique determined that (gamma-CyD)(2)/C(60) (CDF0) efficiently produced singlet oxygen. The rate of singlet oxygen production decreased with increased aggregation, with no production by the fully aggregated sample formed after 150 min of heating (CDF150). UVA irradiation of HLE B-3 in the presence of (gamma-CyD)(2)/C(60) resulted in a significant rise in intracellular protein-derived peroxides. The singlet oxygen quenchers sodium azide and histidine each significantly protected lens cells against (gamma-CyD)(2)/C(60) photodamage, but lutein and Trolox (vitamin E) did not. Clearly, singlet oxygen is an important intermediate in the phototoxicity of monomeric (gamma-CyD)(2)/fullerene. Our results also demonstrate that UVA-blocking sunglasses can limit the ocular phototoxicity of this nanomaterial, while nontoxic endogenous antioxidants like lutein or Trolox cannot provide adequate protection. PMID- 19281133 TI - Highly enantioselective direct Michael addition of nitroalkanes to nitroalkenes catalyzed by amine-thiourea bearing multiple hydrogen-bonding donors. AB - A highly diastereoselective and enantioselective Michael addition of nitroalkanes to nitroalkenes has been achieved by chiral bifunctional amine-thiourea catalyst bearing multiple hydrogen-bonding donors. This catalytic system performs well over a broad scope of substrates, furnishing various 1,3-dinitro compounds in high diastereoselectivity (up to 98:2) and excellent enantioselectivity (up to 99% ee) under mild conditions. Multiple hydrogen bonding donors play a significant role in accelerating reactions, improving diastereoselectivities and enantioselectivities. PMID- 19281134 TI - Isolation of antipodal (-)-versicolamide B and notoamides L-N from a marine derived Aspergillus sp. AB - Antipodal (-)-versicolamide B and notoamides L-N were isolated from a marine derived Aspergillus sp. The possible biosynthetic pathway of enantiomeric pairs of notoamide B and versicolamide B are proposed. Notoamide L is the first metabolite containing 25 carbons in the related prenylated indole alkaloids. Notoamide M is potentially a precursor to the proposed azadiene species involved in the putative intramolecular Diels-Alder reaction in the biogenesis of the bicyclo[2.2.2]diazaoctane ring system. PMID- 19281135 TI - Beta-glycosidation of sterically hindered alcohols. AB - The 2-chloro-2-methylpropanoic ester serves as a steering group in the Schmidt glycosidation reaction. Rapid and efficient glycosidation of a range of sterically hindered alcohols takes place under mild, acidic conditions to afford the glycoside products in high yield and beta-selectivity and without formation of orthoester side products. The 2-chloro-2-methylpropanoic ester is readily cleaved under mild, basic conditions. PMID- 19281136 TI - Efficient Rh(I)-catalyzed direct arylation and alkenylation of arene C-H bonds via decarbonylation of benzoic and cinnamic anhydrides. AB - Efficient rhodium(I)-catalyzed regioselective direct arylation and alkenylation of aromatic C-H bonds has been realized with aromatic carboxylic and cinnamic anhydrides as the coupling partners via decarbonylation and C-H activation under phosphine-free conditions. PMID- 19281137 TI - Synthetic studies on siphonariid polypropionates: synthesis and isomerization of the caloundrin B trioxaadamantane ring system. AB - (1R,3R,5R,7R,8S,9R,10S)-3,5-Diethyl-8,9,10-trimethyl-2,4,6 trioxatricyclo[3.3.1.1(3,7)]decan-1-ol (II), a model of the trioxaadamantane ring system embedded in caloundrin B, was prepared by isomerization of the thermodynamically unstable (9S)-diastereomer (I) in the presence of imidazole. Alternatively, isomerization of I with HF.py or DBU gave the hemiacetal III or its retro-Claisen ester IV, respectively, which represent structural motifs present in the closely related siphonariid polypropionates siphonarin B and baconipyrone C. PMID- 19281138 TI - Synthesis and reaction of DNA oligomers containing modified cytosines related to bisulfite sequencing. AB - The synthesis of DNA oligomers containing N(4)-hydroxy-5,6-dihydrocytosine-6 sulfonate by using ligand-induced base flipping of cytosine followed by the simultaneous addition of bisulfite and hydroxylamine is reported. In contrast to C, the flipped-out 5-methylcytosine was selectively oxidized over thymines and cytosines in the duplex by potassium permanganate. Ligand-induced base flipping is a convenient and powerful strategy for the synthesis of modified cytosines and 5-methylcytosines related to bisulfite sequencing at the predetermined site of DNA. PMID- 19281140 TI - Metal speciation dynamics in soft colloidal ligand suspensions. Electrostatic and site distribution aspects. AB - The theory for metal speciation dynamics in dilute, monodisperse suspensions of spherical core-shell colloidal ligand particles is extended with the impact of the electric double layer (EDL) field and inhomogeneous site distribution inside the particle. The latter is defined by a diffuse, radial distribution for the density of charged polymer segments supporting the ligands L. The site distribution at the scale of the particle suspension and within the colloidal shell results in association/dissociation rate constants (denoted as k(a)* and k(d)*, respectively) that may significantly differ from their homogeneous solution counterparts (k(a) and k(d)). The differences arise from intertwined kinetics of metal-ligand (ML) complex formation/dissociation in the particle shell and diffusive transport of free metal ions M within/outside the shell in the electric field set up by the EDL at the core-shell/electrolyte interphase. The relationship between k(a,d)* and k(a,d) is derived from the numerical evaluation of the spatial, time-dependent distributions of free and bound metal as governed by coupled Nernst-Planck equations corrected by appropriate chemical source term and written in a Kuwabara cell geometry. The average interphasial electrostatic field stemming from the formation of the EDL at the complexing colloidal interphase is obtained from the solution of the nonlinear Poisson Boltzmann equation. The EDL composition is exclusively governed by ions from indifferent background electrolyte present in large excess over free metal species M. The dependences of k(a,d)* on rate constants k(a,d), geometrical details of particle, particle charge, concentration of indifferent background electrolyte, and ligand distribution within the shell are thoroughly discussed within the context of dynamic features for colloidal complex systems. Examination of the chemical equilibrium regime allows addressing explicitly the impact of electrostatics on colloidal complex stability (polyelectrolyte effect). The numerical study is further supported by an approximate analytical expression based on Donnan partitioning and valid under the quasi-steady-state approximation (nonequilibrium chemical regime). The analysis covers the limiting cases of charged rigid particles where binding sites are located at the very surface of the core (e.g., functionalized latex colloids) and polyelectrolyte particles devoid of a hard core (e.g., polysaccharide macromolecules, gel particles). PMID- 19281141 TI - Ionization energies of niobium carbide clusters Nb(n)C(m) (n = 3-10, m = 0-7). AB - We prepared niobium carbide clusters, Nb(n)C(m), in the gas phase by a double laser ablation technique. A photoionization efficiency was measured as a function of the wavelength of an ionization laser to determine ionization energies (E(i)'s) of Nb(n)C(m) (n = 3-10, m = 0-7). The E(i)'s of Nb(4)C(4) and Nb(5)C(3) are found to be the lowest in the clusters studied. When the experimental E(i)'s are compared with the E(i)'s estimated by the density functional calculation previously reported (Harris, H.; Dance, I. J. Phys. Chem. A 2001, 105, 3340 3358), we determined Nb(5)C(3) to have a similar 2 x 2 x 2 cubic structure as Nb(4)C(4). We also found that the E(i)'s of carbon-rich clusters, Nb(n)C(m) (n < or = m), tend to be higher (> 5 eV) than the E(i)'s of the niobium-rich clusters. The high E(i)'s are due to the structure of the carbon-rich clusters: carbon carbon bonding is preferred when the number of carbon atoms exceeds the number of metal atoms. The structure was also discussed in relation to the reactivity of cluster ions with a hydrogen molecule. PMID- 19281142 TI - State-to-state quantum dynamical study of the N + OH --> NO + H reaction. AB - We have studied the quantum dynamics of the N + OH --> NO + H reaction for collision energies up to 0.7 eV. The hyperspherical method has been used in a time-independent formalism. State-to-state reaction probabilities for a total angular momentum J = 0 have been computed. The results show a high reactivity below 0.45 eV and a very small one above this collision energy. Rotational and vibrational product distributions are presented for three collision energies (0.05, 0.1, and 0.5 eV). The vibrational distributions are found to be noninverted at 0.1 eV and inverted peaking at other energies. Rotational distributions are rather hot even if some low rotational states are strongly populated. These features are consistent with both direct and indirect reaction mechanisms. PMID- 19281143 TI - Proton switch correlated with the morphological development of the hydrogen-bond network in H+ (MeOH)m(H2O)1 (m = 1-9): a theoretical and infrared spectroscopic study. AB - A surprising switch of the protonated site from methanol to water in protonated methanol-water mixed clusters, H(+)(MeOH)(m)(H(2)O)(1) (m = 1-9), was investigated by a joint theoretical and vibrational spectroscopic study. Extensive density functional calculations on all possible structural isomers revealed that the switch of the ion core is correlated with the size dependence and structural development of the hydrogen-bond network: (1) the CH(3)OH(2)(+) ion core is preferred for the small-sized clusters of m = 1 and 2, (2) coexistence of the H(3)O(+) and CH(3)OH(2)(+) ion cores is highly plausible for 3 < or = m < or = 7 clusters, and (3) obvious preference of the H(3)O(+) ion core appears from m > or = 8 with the appearance of the characteristic "tricyclic" structure of the hydrogen-bond network. The ion core switch at m approximately 8 is experimentally supported by the infrared photodissociation spectra of the size selected clusters and the size dependence of the fragmentation channel following vibrational excitation. PMID- 19281144 TI - (12)C2H2-Ar van der Waals complex. AB - New theoretical and experimental results on the acetylene-Ar van der Waals complex are presented and the literature is reviewed. New ab initio calculations at the MP2 level were performed using large basis sets with diffuse functions and taking into account the basis set superposition error. It was found that the structure of acetylene is not significantly altered by the complexation and that its vibrational frequencies are only slightly lowered. Finally, it was observed that the calculated properties of the complex (structure, vibrational spectrum, bond dissociation energy) are not sensitive to the structure imposed on acetylene. Experimentally, acetylene-Ar was produced in a supersonic expansion under experimental conditions corresponding to 9 K rotational temperature. Thanks to the performances of CW-CRDS detection, the K(a) = 0 <-- 1, 1 <-- 0, and 2 <-- 1 sub-bands of the nu(1) + nu(3) band could be recorded and resolved and most of their lines assigned. Upper-state rotational constants were fitted, however not including the upper K(a) = 2 state, which shows K-doubling the opposite of the expected. The Lorentzian width of most line profiles sets the mean lifetime to some 7.5 ns. Local perturbations affecting line positions and/or line widths are demonstrated. Additional series of lines tentatively attributed to acetylene-Ar are discussed. PMID- 19281145 TI - Binding motifs for lanthanide hydrides: a combined experimental and theoretical study of the MH(x)(H2)(y) species (M = La-Gd; x = 1-4; y = 0-6). AB - The results of a combined spectroscopic and computational study of lanthanide hydrides with the general formula MH(x)(H(2))(y), where M = La, Ce, Pr, Nd, Sm, Eu, and Gd, x = 1-4, and y = 0-6 are reported. To understand the nature of the dihydrogen complexes formed with lanthanide metal hydride molecules, we have first identified the binary MH(x) species formed in the ablation/deposition process and then analyzed the dihydrogen supercomplexes, MH(x)(H(2))(y). Our investigation shows that the trihydrides bind dihydrogen more weakly than the dihydrides and that the interaction between the central lanthanide and the H(2) molecules occurs via a 6s electron transfer from the lanthanide to the H(2) molecules. Evidence is also presented for the SmH and EuH diatomic molecules and the tetrahydride anions in solid hydrogen. PMID- 19281146 TI - Kinetic barriers of H-atom transfer reactions in alkyl, allylic, and oxoallylic radicals as calculated by composite ab initio methods. AB - Composite ab initio and density functional theory (DFT) methods were used to explore internal hydrogen-atom transfers in a variety of primary, secondary, and tertiary alkyl and functionalized radicals with implications for combustion environments. The composite ab initio method G3MP2B3 was found to achieve the most reasonable balance between accuracy and economy in modeling the energetics of these reactions. Increased alkyl substitution reduced barriers to isomerization by about 10 and 20 kJ mol(-1) for secondary and tertiary radical formation, respectively, relative to primary radical reactions and was relatively insensitive to the transition-state ring size (extent of H-atom internal shift). Reactions involving alkenyl and alkanoyl radicals were also explored. Hydrogen atom transfers involving allylic radical formation demonstrated barrier heights that were 15-20 kJ mol(-1) lower than those in corresponding alkyl radicals, whereas those involving oxoallylic species (alpha-site radicals of aldehydes and ketones) were 20-40 kJ mol(-1) lower. In the cases of the alkyl radicals, enthalpies of activation were seen to scale with enthalpies of reaction. This correlation was not seen, however, in the cases of the allylic and oxoallylic radicals; this fact has significant implications in combustion chemistry and mechanism development, considering that such Evans-Polanyi correlations are widely used in estimating barrier heights for rate expressions. PMID- 19281147 TI - Density functional theory study on the semiconducting properties of metal phthalocyanine compounds: effect of axially coordinated ligand. AB - To investigate the effect of axially coordinated ligand(s) on the semidconducting properties of metal phthalocyanine complexes, density functional theory (DFT) calculations were carried out in terms of the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energy, ionization energy (IE), electronic affinity (EA), and reorganization energy (lambda) of F(2)SnPc, Cl(2)SnPc, I(2)SnPc, OSnPc, OVPc, and Cl(2)TiPc. For the purpose of comparative studies, calculation on SnPc without axially coordinated ligand has also been conducted. The electronic couplings (V) and the charge transfer mobilities for the electron of metal phthalocyanine compounds with reported single crystal structures for Cl(2)SnPc, I(2)SnPc, and Cl(2)TiPc are also calculated. Comparison of the calculated results of SnPc with F(2)SnPc, Cl(2)SnPc, I(2)SnPc, and OSnPc indicates that introduction of axially coordinated ligand(s) obviously lowers the HOMO and LUMO energies of metal phthalocyanine complexes but does not change their energy difference, which results in an increase in their electronic affinity and ionization energy for metal phthalocyanine complexes containing axially coordinated ligand(s). This result is responsible for the decrease in the electron injection barrier and increase in the hole injection barrier of metal phthalocyanine complexes containing axially coordinated ligand(s) in comparison with metal phthalocyanine complexes without axially coordinated ligand, leading to the change in the nature of semiconductivity from p-type for SnPc to n-type for F(2)SnPc, Cl(2)SnPc, I(2)SnPc, and OSnPc. Because of the smaller electronegativity of V(IV) than that of Sn(IV), OVPc is revealed to display p type semiconductivity in terms of electronic affinity (EA(v)). In contrast, Cl(2)TiPc is revealed to show n-type semiconductivity because of its large electronic affinity (EA(v)). The present work, representing the first theoretical effort toward understanding the effect of axially coordinated ligand(s) on the semiconducting properties of metal phthalocyanine complexes, will be helpful for designing and preparing novel phthalocyanine semidconductors with good organic field effect transistor (OFET) performance. PMID- 19281148 TI - Improper hydrogen-bonding CH x Y interactions in binary methanol systems as studied by FTIR and Raman spectroscopy. AB - Fourier Transform infrared spectroscopy and Raman spectroscopy have been used to investigate hydrogen bonding of methanol in different solvents with an aim to explore potential experimental evidence for improper hydrogen bonding involving the methyl group of methanol as suggested by various computational studies. Pure methanol and solutions of methanol in water, acetonitrile, carbon tetrachloride, deuterium oxide, and deuterated acetonitrile have been studied over a range of concentrations. Wavenumber shifts of the CH stretching vibrations were examined to determine if the CH from methanol participates in hydrogen bonding. New concepts of the vibrational wavenumber and integrated intensity at infinite dilution are proposed and given the respective symbols nu(CH(o)) and C(j,CH)*(o). Using the results obtained for methanol in carbon tetrachloride as a reference, shifts in nu(CH(o)) of methanol to higher wavenumbers (blue shifts) were observed in each of the other solvents studied, with the shifts being greatest for the methanol-water interactions. The shifts in vibrational wavenumber suggest possible improper hydrogen bonding, although at this stage a definitive conclusion is not possible. The C(j,CH)*(o) results show that there is no distinguishable change in the methanol CH stretch integrated intensity in carbon tetrachloride and acetonitrile, while there is a significant decrease in the methanol CH stretch integrated intensity in the water solutions. PMID- 19281149 TI - PEGylated viral nanoparticles for biomedicine: the impact of PEG chain length on VNP cell interactions in vitro and ex vivo. AB - PEGylation is an effective strategy for reducing biospecific interactions for pharmaceuticals. The plant virus Cowpea mosaic virus (CPMV) has been studied for potential nanobiomedical applications by virtue of its natural interactions with mammalian endothelial cells. To investigate the degree of PEGylation required to retarget CPMV-based formulations to other destinations, two CPMV-PEG formulations, CPMV-PEG1000 (P1) and CPMV-PEG2000 (P2) were tested. Modeling suggested that the PEG chains were displayed as flattened mushrooms on the particle with an estimated surface grafting area of 0.53% for P1 and 0.83% for P2. Only the P2 formulation effectively shielded the particles from interacting with cells or tissues, suggesting that either key interacting regions on the particle surface were blocked or that a sufficient hydration shell had been generated to inhibit cellular interactions. The large CPMV surface area available after PEGylation allows further attachment of imaging and therapeutic molecules to the particle to generate multifunctionality. PMID- 19281150 TI - Poly(ortho ester amides): acid-labile temperature-responsive copolymers for potential biomedical applications. AB - A new, convenient pathway is developed to synthesize highly hydrolytically labile poly(ortho ester amide) (POEA) copolymers that overcomes some of the major weaknesses of the traditional methods of synthesizing poly(ortho esters) and their derivatives. A diamine monomer containing a built-in, stabilized ortho ester group was synthesized and was used for polycondensation with diacid esters, giving rise to a series of POEA copolymers with unique stimuli-responsive properties. The POEA undergoes temperature-responsive, reversible sol-gel phase transition in water. Phase diagrams of the POEA/H(2)O mixture reveal the concentration-dependent existence of different phases, including hydrogel and opaque or clear solution. Such behavior may be attributed to the temperature dependent hydrogen-bonding involving the amide groups in the POEA backbone and hydrophobic interactions between POEA chains, and it is tunable by selecting diacid monomers with different chemical structures. The kinetics of POEA mass loss in physiological aqueous buffers and release of a model macromolecular drug, fluorescently labeled dextran, are nearly zero-order, suggesting predominantly surface-restricted polymer erosion. The rates of polymer erosion and drug release are much faster at pH 5.0 than pH 7.4. No cytotoxicity was found for the polymer extracts and the polymer degradation products at concentrations as high as 1 mg/mL. The normal morphology of fibroblasts cultured directly in contact with POEA films was not altered. These novel acid-labile temperature-responsive POEA copolymers may be potentially useful for a wide range of biomedical applications such as minimal invasive delivery of controlled-release drug formulations that respond to biological temperature and acidic-pH environments in cells and tissues. PMID- 19281151 TI - Asymmetric poly(ethylene glycol) star polymers with a cholic acid core and their aggregation properties. AB - Poly(ethylene glycol) (PEG) arms are grafted onto a cholic acid core via anionic polymerization, yielding star-shaped polymers with a unique asymmetric structure with facial amphiphilicity. Well-defined cholic acid-PEG(4) stars (polydispersity index, ca. 1.05) with tunable molar masses (ca. 1000-13,000) were obtained and characterized by the use of size exclusion chromatography, MALDI-TOF mass spectrometry, NMR spectroscopy, and thermal analysis. The asymmetric star polymers were found to aggregate differently from cholic acid salt. The critical aggregation concentrations of the star polymers were determined by surface tension measurements, and spherical aggregates of the polymers with different PEG chain lengths were observed by transmission electron microscopy using the freeze fracture etching technique. The elongated aggregates formed by the sodium salt of cholic acid were also observed. The hydrodynamic diameters of the aggregates were also measured using dynamic light scattering technique. The formation of aggregates makes them interesting systems as potential drug carriers. PMID- 19281152 TI - Fatty acid-derived diisocyanate and biobased polyurethane produced from vegetable oil: synthesis, polymerization, and characterization. AB - A new linear saturated terminal diisocyanate was synthesized from oleic acid via Curtius rearrangement, and its chemical structure was identified by FTIR, (1)H and (13)C NMR, and MS. The feasibility of utilizing this new diisocyanate for the production of polyurethanes (PUs) was demonstrated by reacting it with commercial petroleum-derived polyols and canola oil-derived polyols, respectively. The physical properties of the PUs prepared from fatty acid-derived diisocyanate were compared to those prepared from the same polyols with a similar but petroleum derived commercially available diisocyanate: 1,6-hexamethylene diisocyanate. It was found that the fatty acid-derived diisocyanate was capable of producing PUs with comparable properties within acceptable tolerances. This work is the first that establishes the production of linear saturated terminal diisocyanate derived from fatty acids and corresponding PUs mostly from lipid feedstock. PMID- 19281153 TI - Multiresponsive hybrid microgels and hollow capsules with a layered structure. AB - Various stimuli-responsive composite particles with a high control of their internal structure and their corresponding hollow capsules are synthesized and characterized by photon correlation spectroscopy, TEM, and AFM. Core-shell particles with a silica core and a thermoresponsive shell are obtained by polymerization of N-isopropylacrylamide (NIPAM) in the presence of silica seeds grafted with a high density of gamma-methacryloxypropyltrimethoxysilane (MPS). The influence of the synthesis conditions is studied. The shell thickness increases when the monomer concentration increases in a limited range where uniform composite particles with a single core are obtained. At constant monomer concentration, the shell thickness does not depend on the size of the silica seeds, but the presence of free unbound microgels is observed when the silica surface area decreases. A range of particle diameters and shell thicknesses is thus obtained, which can lead to the corresponding hollow capsules by exposure to hydrofluoric acid solution. The volume phase transition temperature of these materials can be easily tuned by replacing the NIPAM monomer by another N alkylacrylamide derivative. However, the incorporation of comonomers such as acrylic acid (AA) and a phenylboronic acid (PBA) derivative inhibits the formation of core-shell structures. In order to get pH or glucose responsiveness, these functional groups can be incorporated in the outer shell of a core-double shell structure, with pNIPAM as intermediate shell. pH-responsive and glucose responsive composite particles are obtained by this method with a high control of their internal structure. PMID- 19281155 TI - Controlling oligonucleotide surface density in light-directed DNA array fabrication. AB - Over the past two decades high-density DNA arrays have developed into a central technology for nucleic acid analyses. Important application areas include whole genome gene expression studies, high throughput analyses of single nucleotide polymorphisms, and, most recently, the determination of binding site specificities for transcription factors and other critical elements involved in gene regulation. A key parameter in the performance of DNA arrays is the density of the surface-bound oligonucleotides, which strongly affects both thermodynamic and kinetic aspects of DNA hybridization. In this report, we describe an approach for the control of oligonucleotide density in photolithographically fabricated DNA arrays, based upon a controlled UV light deprotection procedure. Modulation of the UV exposure permits a desired degree of deprotection of surface synthesis sites; a subsequent capping reaction to inactivate the exposed sites leaves only a desired fraction of active sites remaining for synthesis, corresponding to a lower oligonucleotide density. It is shown that the procedure is reasonably general, in that it is readily transferable to alternative substrate materials with similar results. PMID- 19281156 TI - Using light to guide the self-sustained motion of active gels. AB - We undertake the first computational study to determine the effect of light on polymer gels undergoing the Belousov-Zhabotinsky (BZ) reaction. The BZ gels are unique materials because they can undergo rhythmic mechanical oscillations in the absence of external stimuli. The BZ reaction, however, is photosensitive. Via simulations, we demonstrate that the interplay between the chemoresponsive gels and the photosensitive reaction can cause millimeter sized BZ gels to exhibit autonomous, directed motion or reorientation away from 4 the light. In effect, we show that these synthetic BZ "worms" display a fundamental biomimetic behavior: movement away from an adverse environmental condition, which in the context of the BZ reaction is the presence of light. PMID- 19281157 TI - Superhydrophobic conductive carbon nanotube coatings for steel. AB - We report the synthesis of superhydrophobic coatings for steel using carbon nanotube (CNT)-mesh structures. The CNT coating maintains its structural integrity and superhydrophobicity even after exposure to extreme thermal stresses and has excellent thermal and electrical properties. The coating can also be reinforced by optimally impregnating the CNT-mesh structure with cross-linked polymers without significantly compromising on superhydrophobicity and electrical conductivity. These superhydrophobic conductive coatings on steel, which is an important structural material, open up possibilities for many new applications in the areas of heat transfer, solar panels, transport of fluids, nonwetting and nonfouling surfaces, temperature resilient coatings, composites, water-walking robots, and naval applications. PMID- 19281158 TI - From spherical to polymorphous dispersed phase transition in water/oil emulsions. AB - Optical scanning tomography is used to characterize bulk properties of transparent water-in-paraffin oil emulsions stabilized with hexadecyl trimethylammonium bromide (CTAB) and silica nanoparticles. A flow of 500 hundred images is used to analyze each scanning shot with a precision of about 1 microm. The role of silica particles in the shape of the water droplets is investigated. Depending on the concentration of CTAB and silica nanoparticles, a transition occurs in their geometry that changes from spherical to polymorphous. This transition is controlled by the ratio R=[CTAB]/[SiO2] and is described using an identification procedure of the topology of the gray level contours of the tomographic images. The transition occurs for Rcrit approximately 3x10(-2) and is shown to correspond to a pH of the dispersed phase of 8.5. PMID- 19281159 TI - Proton absorption in as-synthesized mesoporous silica nanoparticles as a structure-function relationship probing mechanism. AB - A new method to investigate the effect of pore geometry on diffusion processes in mesoporous silica nanoparticles and other types of micro- and mesoporous structures is put forward. The method is based on the study of proton diffusion from a liquid surrounding the mesoporous particles into the particle pore system. The proton diffusion properties are assessed for a variety of as-synthesized mesoporous nano- and microparticles with two-dimensional and three-dimensional connectivity. Results show that the diffusion coefficients are higher for the proton absorption process than for the release of surfactant template molecules, and that they overall follow the same trend with the more complex three dimensional mesocaged particles showing the highest diffusion coefficients. The pore geometry (cylindrical pores versus cage-type pores) and structure connectivity are found to play a key role for the effects observed. The results put forward in the present work should offer a valuable tool in the development of porous nanomaterials in a range of applications including the use as catalysis and separation enhancers in the petrochemical industry, as scaffolds for hydrogen storage, and as drug delivery vehicles for sustained release and gene transfection. PMID- 19281160 TI - Ring-opening polymerization-mediated controlled formulation of polylactide-drug nanoparticles. AB - We report here a unique method for formulating doxorubicin-polylactide (Doxo-PLA) conjugate nanoparticles, known as nanoconjugates (NCs), through Doxo/(BDI)ZnN(TMS)(2)-mediated [(BDI) = 2-((2,6-diisopropylphenyl)amido)-4-((2,6 diisopropylphenyl)-imino)-2-pentene], chemo- and regioselective polymerizations of lactide (LA) followed by nanoprecipitation. When Doxo/(BDI)ZnN(TMS)(2) was mixed with 1-pyrenemethanol (Pyr-OH) and 1-pyrenemethylamine (Pyr-NH(2)) and the mixture was utilized for the polymerization of LA, remarkable chemoselectivity was observed. Pyr-OH was completely consumed and covalently linked to the terminus of the PLA, whereas the Pyr-NH(2) remained intact in the polymerization solution. When Doxo was used as the initiator to polymerize LA in the presence of (BDI)ZnN(TMS)(2), the polymerization was complete within hours, with nearly 100% Doxo-loading efficiency and 100% LA conversion. Doxo loading as high as 27% could be achieved at a LA/Doxo ratio of 10. Both the steric bulk of the chelating ligand and the metal catalyst had dramatic effects on the regioselectivity during the initiation step. When Doxo/(BDI)ZnN(TMS)(2) was mixed with succinic anhydride (SA) to mimic the initiation of Doxo/(BDI)ZnN(TMS)(2)-mediated LA polymerization, Doxo-14-succinic ester (Doxo-SE) was the predominate product. When the steric bulk of BDI was reduced or when the BDI ligand was removed, significant amounts of Doxo-4',14-bis-succinic ester (Doxo-2SE) and Doxo-4',9,14-trisuccinic ester (Doxo-3SE) were formed. The use of (BDI)MgN(TMS)(2) in such a reaction also resulted in reduced regioselectivity and formation of both Doxo-SE and Doxo-2SE. Doxo/(BDI)ZnN(TMS)(2)-mediated LA polymerizations yielded Doxo-PLA conjugates with well-controlled molecular weights and polydispersities (as low as 1.02). The nanoprecipitation of Doxo-PLA formed NCs less than 150 nm in size with narrow particle size distributions. The sustained release of Doxo from Doxo-PLA NCs was achieved without a burst release. This method may have widespread utility for controlled conjugation of hydroxyl-containing agents to polyesters and formation of corresponding nanoparticles. PMID- 19281161 TI - Inter- and intramolecular [4 + 3] cycloadditions using epoxy enol silanes as functionalized oxyallyl cation precursors. AB - Using epoxy enol triethylsilanes as oxyallyl cation precursors, [4 + 3] cycloadditions with various dienes occur under catalysis by silyl triflates and acids in good yields. The intramolecular [4 + 3] cycloaddition proceeds under mild conditions and generate hydroxylated cycloadducts with high diastereoselectivity and yields. Enantiomerically pure epoxy enol silanes have been shown to give excellent yields of the optically pure cycloadduct bearing multiple stereocenters. PMID- 19281162 TI - Intramolecular rhodium-catalyzed [2+2+2] cyclizations of diynes with enones. AB - The Rh(I)-catalyzed inter- and intramolecular [2+2+2] cyclization of diynes with alpha,beta-unsaturated enones proceeds with microwave promotion in good yields. This chemistry was applied to the synthesis of (-)-alcyopterosin I. PMID- 19281164 TI - Demonstration of a peroxide shunt in the tetrahydropterin-dependent aromatic amino acid monooxygenases. AB - The nonheme iron enzyme phenylalanine hydroxylase, tyrosine hydroxylase, and tryptophan hydroxylase catalyze the hydroxylation of their aromatic amino acid substrates using a tetrahydropterin as the source of electrons. The hydroxylating intermediate is proposed to be an Fe(IV)O species. We report here that all three enzymes will catalyze hydroxylation reactions using H(2)O(2) in place of tetrahydropterin and oxygen, forming tyrosine and 3-hydroxyphenylalanine from phenylalanine, 4-HOCH(2)-phenylalanine from 4-CH(3)-phenylalanine, and hydroxycyclohexylalanine from 3-cyclohexylalanine. No peroxide-dependent reaction is seen with active site mutants of TyrH and PheH in which the stability or reactivity of the iron center is compromised. These results provide further support for an Fe(IV)O hydroxylating intermediate. PMID- 19281165 TI - Molecular orientation and field-effect transistors of a rigid rod conjugated polymer thin films. AB - Molecular orientation in thin films of a rigid rod conjugated polymer, a derivative of poly(para-phenylene ethynylene)s with linear side chains and thioacetyl end groups, was investigated by reflection-absorption infrared spectroscopy and X-ray diffraction technique. The results indicated that TA-PPE molecules tended to align with their backbone planes perpendicular to substrates, that is, with an "edge-on" molecular orientation in the films. Such molecular orientation is favorable for the efficient carrier transport in two-dimensional direction in the polymer films (i.e., via both the intrachain and interchain), so that high performance organic field-effect transistors were fabricated with hole mobility at around approximately 4.3 x 10(-3) cm(2)/Vs. PMID- 19281166 TI - Assembly and stabilization of multi-amino acid zwitterions by the Zn(II) ion: a computational exploration. AB - Density functional calculations were employed to stabilize multi-amino acid zwitterions by metal ions as well as to understand the stabilizing mechanism. This work distinguishes from the previous metalations and stabilizations of a single amino acid zwitterion, where the metal ions are coordination-unsaturated. It was found that up to three Gly zwitterions can be stabilized by the Zn(2+) ion at the same time. The addition of the fourth Gly zwitterion to the Zn(Gly)(3) system causes the spontaneous transformation of one zwitterion to the neutral isomer. The single mutations of Gly in Zn(Gly)(4) with Arg or Asp render the Zn(2+)-combined four amino acid zwitterions geometrically stable. The double mutation of Gly with Arg further stabilizes the structure with four amino acid zwitterions, with its energy being lower than the one containing one neutral and three zwitterionic isomers. The systematic studies on a series of M-combined amino acid systems (M = Zn(2+), Zn(+), Li(+), Na(+)) indicated that enough charge transfers from the metal ions are essential to stabilize the amino acid zwitterions. There exist critical values of charge transfers, and for Zn(2+) combined amino acid systems, an estimation of 0.20 |e| is required to stabilize each Gly zwitterion. Further analysis revealed that the charge transfers rather than the formations of direct M-O bonds play a more important role in the stabilizations of amino acid zwitterions. PMID- 19281167 TI - Hybrid coarse-graining approach for lipid bilayers at large length and time scales. AB - A hybrid analytic-systematic (HAS) coarse-grained (CG) lipid model is developed and employed in a large-scale simulation of a liposome. The methodology is termed hybrid analytic-systematic because one component of the interaction between CG sites is variationally determined from the multiscale coarse-graining (MS-CG) methodology, whereas the remaining component utilizes an analytic potential. The systematic component models the in-plane center-of-mass interaction of the lipids as determined from an atomistic-level MD simulation of a bilayer. The analytic component is based on the well-known Gay-Berne ellipsoid-of-revolution liquid crystal model and is designed to model the highly anisotropic interactions at a highly coarse-grained level. The HAS CG approach is the first step in an "aggressive" CG methodology designed to model multicomponent biological membranes at very large length and time scales. PMID- 19281168 TI - How is the reactivity of cytochrome P450cam affected by Thr252X mutation? A QM/MM study for X = serine, valine, alanine, glycine. AB - Proton transfer reactions play a vital role in the catalytic cycle of cytochrome P450cam and are responsible for the formation of the iron-oxo species called Compound I (Cpd I) that is supposed to be the active oxidant. Depending on the course of the proton transfer, protonation of the last observable intermediate (ferric hydroperoxo complex, Cpd 0) can lead to either the formation of Cpd I (coupling reaction) or the ferric resting state (uncoupling reaction). The ratio of these two processes is drastically affected by mutation of the Thr252 residue. In this work, we study the effect of Thr252X (X = serine, valine, alanine, glycine) mutations on the formation of Cpd I by means of hybrid quantum mechanical/molecular mechanical (QM/MM) calculations and classical simulations. In the wild-type enzyme, the coupling reaction is favored since its rate-limiting barrier is 13 kcal/mol lower than that for uncoupling. This difference is reduced to 7 kcal/mol in the serine mutant. In the case of valine, alanine, and glycine mutants, an additional water molecule enters the active site and lowers the activation energy of the uncoupling reaction significantly. With the additional water molecule, coupling and uncoupling have similar barriers in the valine mutant, and the uncoupling reaction becomes favored in the alanine and glycine mutants. These findings agree very well with experimental results and thus confirm the assumption that uncontrolled proton delivery by solvent water networks is responsible for the uncoupling reaction. The present study provides a detailed mechanistic understanding of the role of the Thr252 residue. PMID- 19281169 TI - Explaining the differential solubility of flue gas components in ionic liquids from first-principle calculations. AB - Flue gas is greatly responsible for acid rain formation and global warming. New generation ionic liquids (ILs) have potential in controlling the flue gas emissions, as they acquire high absorptivity for the component gases SO(2), CO(2), etc. The association of the IL-gas interactions to the absorptivity of gas molecules in ILs is, however, poorly understood. In this paper, we present a molecular level description of the interactions of ILs with SO(2), CO(2), and N(2) and show its implications to the differential gas solubility. Our results indicate that the IL anion-gas interactions play a key role in deciding the gas solubility in ILs, particularly for polar gases such as SO(2). On the other hand, regular solution assumption applies to N(2) solubility. In accordance with the previous theoretical and experimental findings, our results also imply that the IL anions dominate the interactions with gas molecules while the cations play a secondary role and the underlying fluid structures of the ILs remain unperturbed by the addition of gas molecules. PMID- 19281170 TI - Matrix-assisted laser desorption ionization imaging mass spectrometry detection of a magnetic resonance imaging contrast agent in mouse liver. AB - The present paper describes the detection of a magnetic resonance imaging (MRI) contrast agent by matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS). The contrast agent was analyzed in both frozen and paraformaldehyde-fixed mouse livers explanted after its in vivo administration, and its identity was confirmed by fragmentation experiments. Moreover, a semiquantitative analysis was performed, evaluating its content in livers from mice sacrificed at different postadministration times. To the best of our knowledge, this is the first description of a MALDI-IMS analysis of MRI contrast agents and the first time that results obtained by MALDI-IMS are validated by both an in vivo (MRI) and an ex vivo (inductively coupled plasma atomic emission spectroscopy, ICP-AES) technique. Results shown in the present paper demonstrate the possibility of using MALDI-IMS for drug biodistribution analysis. Obviously, this application is particularly interesting in the case of unlabeled compounds, which cannot be detected by any of the other imaging techniques. PMID- 19281171 TI - Structural analysis of an open active site conformation of nonheme iron halogenase CytC3. AB - CytC3, a member of the recently discovered class of nonheme Fe(II) and alpha ketoglutarate (alphaKG)-dependent halogenases, catalyzes the double chlorination of L-2-aminobutyric acid (Aba) to produce a known Streptomyces antibiotic, gamma,gamma-dichloroaminobutyrate. Unlike the majority of the Fe(II)-alphaKG dependent enzymes that catalyze hydroxylation reactions, halogenases catalyze a transfer of halides. To examine the important enzymatic features that discriminate between chlorination and hydroxylation, the crystal structures of CytC3 both with and without alphaKG/Fe(II) have been solved to 2.2 A resolution. These structures capture CytC3 in an open active site conformation, in which no chloride is bound to iron. Comparison of the open conformation of CytC3 with the closed conformation of another nonheme iron halogenase, SyrB2, suggests two important criteria for creating an enzyme-bound Fe-Cl catalyst: (1) the presence of a hydrogen-bonding network between the chloride and surrounding residues, and (2) the presence of a hydrophobic pocket in which the chloride resides. PMID- 19281172 TI - Oxygen ordering and mobility in YBaCo(4)O(7+delta). AB - Extraordinary oxygen ordering and mobility have been found in an oxygen nonstoichiometric mixed-valence cobalt oxide, YBaCo(4)O(8.5). The excess oxygen atoms appear to be incorporated in an orderly way into the YBaCo(4)O(7) parent lattice with different configurations as the oxygen content varies. Intense electron-beam irradiation was used to observe recurrent oxygen migration in the lattice, causing reversible structural modulation transitions. The oxygen migration can be attributed mainly to the electron-beam heating effect. This study highlights the high degree of freedom of surplus oxygen within the investigated structure and advances our understanding of the oxygen diffusion process in related transition-metal oxide systems. PMID- 19281173 TI - Inequivalent contribution of the five tryptophan residues in the C-lobe of human serum transferrin to the fluorescence increase when iron is released. AB - Human serum transferrin (hTF), with two Fe3+ binding lobes, transports iron into cells. Diferric hTF preferentially binds to a specific receptor (TFR) on the surface of cells, and the complex undergoes clathrin dependent receptor-mediated endocytosis. The clathrin-coated vesicle fuses with an endosome where the pH is lowered, facilitating iron release from hTF. On a biologically relevant time scale (2-3 min), the factors critical to iron release include pH, anions, a chelator, and the interaction of hTF with the TFR. Previous work, in which the increase in the intrinsic fluorescence signal was used to monitor iron release from the hTF/TFR complex, established that the TFR significantly enhances the rate of iron release from the C-lobe of hTF. In the current study, the role of the five C-lobe Trp residues in reporting the fluorescence change has been evaluated (+/-sTFR). Only four of the five recombinant Trp --> Phe mutants produced well. A single slow rate constant for iron release is found for the monoferric C-lobe (FeC hTF) and the four Trp mutants in the FeC hTF background. The three Trp residues equivalent to those in the N-lobe differed from the N-lobe and each other in their contributions to the fluorescent signal. Two rate constants are observed for the FeC hTF control and the four Trp mutants in complex with the TFR: k(obsC1) reports conformational changes in the C-lobe initiated by the TFR, and k(obsC2) is ascribed to iron release. Excitation at 295 nm (Trp only) and at 280 nm (Trp and Tyr) reveals interesting and significant differences in the rate constants for the complex. PMID- 19281174 TI - Activation of an alkyl C-H bond geminal to an agostic interaction: an unusual mode of base-induced C-H activation. AB - Deuterium labeling studies indicate that base-induced intramolecular C-H activation in the agostic complex 2-D proceeds with exclusive removal of a proton from the methyl arm of an (i)Pr substituent on the N-heterocyclic carbene (NHC) ligand. Computational studies show that this alkyl C-H bond activation reaction involves deprotonation of one of the C-H bonds that is geminal to the agostic interaction, rather than the agostic C-H bond itself. The reaction is readily accessible at room temperature, and a computed activation barrier of DeltaE (double dagger)(calcd) = +11.8 kcal/mol is found when the NHC 1,3-diisopropyl-4,5 dimethylimidazol-2-ylidene is employed as the external base. Charge analysis reveals that the geminal hydrogens are in fact more acidic than the agostic proton, consistent with their more facile deprotonation. PMID- 19281175 TI - Complementary multianalytical approach to study the distinctive structural features of the main humic fractions in solution: gray humic acid, brown humic acid, and fulvic acid. AB - Previous studies have indicated that the main fractions of humic substances (HS), gray humic acid (GHA), brown humic acid (BHA), and fulvic acid (FA), present different molecular patterns in water solution that are probably associated with specific structural features. However, the techniques used in these previous studies did not permit clarification of the principal qualitative characteristics of these structures. To study more in depth this subject several GHA, BHA, and FA have been analyzed through the complementary use of UV-visible and FTIR spectroscopy, (13)C NMR, thermogravimetry, and pyrolysis GC-MS. The results indicate that the studied humic fractions have different and distinctive structural features. Thus, large and nonpolar structural units (paraffins, olefins, terpenes) and aliphatic structures seem to accumulate in the gray fraction, whereas the smallest and more polar (furfural, phenols) and simpler structural units (sugar- and amino acid-related structures) are present in the fulvic one. BHA has a higher content in polycyclic aromatic moieties, S containing compounds and aromatic structures, thus suggesting the presence of more condensed aromatic rings. Likewise, differences in both the presence of polar groups and the apparent molecular size explain the pattern of solubility as a function of pH and ionic strength (I) that defines each HS fraction. These results also indicate that the structural differences among the HS fractions are not only quantitative (the presence of the same type of structures differing in size and the concentration of functional groups) but also qualitative, because each fraction presented different and distinctive structural domains. These structural domains explain the molecular patterns associated with each HS fraction. Thus, the presence of smaller and more O-functionalized structural units including aromatic domains in FA explain their tendency to form molecular aggregates (hydrogen bridges, metal bridges, and hydrophobic interactions) in solution. This fact could also explain the presence of molecular aggregates in BHA, although to a lesser extent than in FA. Finally, the dominant aliphatic and less functionalized character of GHA may justify its lower tendency to form aggregates in solution at neutral and alkaline pH. Likewise, the results also indicate that the different structural domains associated with these fractions may be the consequence of diverse biosynthetic pathways involving different precursors. PMID- 19281176 TI - Analysis of sparkling wine lees surface volatiles by optimized headspace solid phase microextraction. AB - During sparkling wine aging, a narrow interaction is established between wine and lees of second fermentation, which remain in contact during long periods. In order to contribute to the knowledge on this interaction, volatile compounds retained by lees were described in this study. With this aim, solid phase microextraction (SPME) conditions were optimized in order to increase the sensibility for the analysis of volatiles sorbed by lees. This allowed proving the capacity of sparkling wine lees of second fermentation to retain several volatile compounds related with wine aroma: esters, aldehydes, norisoprenoids and terpenes known for their positive flavor impact were found in lees headspace. Most of them were previously described in "Cava" sparkling wine, while some compounds, such as the tentatively identified trimethyl tetrahydronaphthalenes, were not previously identified in yeasts or wine. PMID- 19281177 TI - Computational studies of load-dependent guest dynamics and free energies of inclusion for CO2 in low-density p-tert-butylcalix[4]arene at loadings up to 2:1. AB - The structure, dynamics, and free energies of absorption of CO(2) by a low density structure (P4/n) of calixarene p-tert-butylalix[4]arene (TBC4) at loadings up to 2:1 CO(2):TBC4 have been studied by using molecular dynamics simulations with two sources of initial TBC4 structures (TBC4-T and TBC4-U). The CO(2)/TBC4 complex structure is very sensitive to the initial lattice spacing of TBC4. From the computed radial distribution functions of CO(2) molecules, a CO(2) dimer is observed for TBC4-T and a cage-interstitial CO(2) structure is suggested for TBC4-U. The dynamics of the CO(2) molecules show little initial TBC4 structural dependency. The free energy of inclusion for a single CO(2) in this TBC4 structure for various loadings is -4.0 kcal/mol at 300 K and -1.8 kcal/mol at 450 K, showing that CO(2) inclusion is favored. The fully loaded 1:1 CO(2):TBC4 system is slightly less favorable at -3.9 and -1.2 kcal/mol at 300 and 450 K, respectively. The first CO(2) added beyond 1:1 loading shows a significant drop in absorption energy to -1.9 and +1.9 kcal/mol at 300 and 450 K. These data are consistent with experimental results showing that low-density structures of TBC4 are able to absorb CO(2) at loadings greater than 1:1 but retention is lower than for 1:1 loaded systems indicating the free energy of inclusion for addition of the CO(2) above 1:1 is less favorable. PMID- 19281178 TI - Binding of Neisseria meningitidis pili to berry polyphenolic fractions. AB - Blocking bacterial adhesion to host surfaces provides novel potential to control infections. The present study was directed to binding and inhibitory activity of different fresh berries and berry and fruit juices against Neisseria meningitidis . Berries and juices were fractionated according to their molecular size into three fractions. A microtiter well assay for binding of N. meningitidis pili to berry and juice fractions was constructed. In addition, adhesion inhibition to human epithelial cells (HEC-1B) was tested. The active fractions were then subfractionated by employing solid-phase extraction. Subfractions were characterized by RP-HPLC-DAD, and the pili binding was evaluated by using microtiter well binding assay. Binding and inhibitory activity were detected to bilberry, cranberry, lingonberry, and crowberry fractions, which contained anthocyanins or a mixture of proanthocyanidins and flavonols. Thus, the findings identify several previously unknown binding and inhibitory activities and may suggest Vaccinium berries and crowberry as promising sources against meningococcal adherence. PMID- 19281179 TI - Calculation of the positron annihilation rate in PsH with the positronic extension of the explicitly correlated nuclear-electronic orbital method. AB - The nuclear-electronic orbital explicitly correlated Hartree-Fock (NEO-XCHF) method is modified and extended to study electron-positron quantum systems. The NEO-XCHF method is more computationally efficient than the explicitly correlated methods previously applied to positron systems because only the electron-positron dynamical correlation is treated explicitly in NEO-XCHF. As a result, the form of the wave function is much simpler with fewer parameters, and the variational optimization of the molecular orbital parameters is performed through an iterative scheme rather than a stochastic optimization. The NEO-XCHF approach is used to calculate the positron annihilation rate for positronium hydride (PsH). The resulting annihilation rate for PsH is within 20% of the most accurate values available and is calculated at a fraction of the computational cost. These results suggest that qualitatively accurate positron annihilation rates can be calculated treating only electron-positron correlation explicitly, leading to significant computational savings by neglecting electron-electron dynamical correlation. Thus, the NEO-XCHF approach could potentially enable the calculation of qualitatively accurate positron annihilation rates for larger positron systems. PMID- 19281180 TI - Degree of initial hole localization/delocalization in ionized water clusters. AB - The electronic structure of ionized bulk liquid water presents a number of theoretical challenges. Not the least of these is the realization that the detailed geometry of the hydrogen bonding network is expected to have a strong effect on the electronic couplings between water molecules and thus the degree of delocalization of the initially ionized system. This problem is approached from a cluster perspective where a high-level coupled cluster description of the electronic structure is still possible. Building on the work and methodology developed for the water dimer cation [J. Phys. Chem. A 2008, 112, 6159], the character and spectrum of electronic states of the water hole and their evolution from the dimer into higher clusters is presented. As the time evolution of the initially formed hole can in principle be followed by the system's transient absorption spectrum, the state spacings and transition strengths are computed. An analysis involving Dyson orbitals is applied and shows a partially delocalized nature of states. The issue of conformation disorder in the hydrogen bonding geometry is addressed for the water dimer cation. PMID- 19281181 TI - Thermally induced interconversions of metal-pyrimidine-4,6-dicarboxylate polymers: a structural, spectroscopic, and magnetic study. AB - Continuing our work on the structural and magnetic aspects of the one-dimensional (1-D) coordination polymers of the [M(pmdc)(H(2)O)(2)] x H(2)O kind (M = Fe, Co, Ni, Cu, Zn; pmdc = pyrimidine-4,6-dicarboxylate), we have combined ab initio X ray powder diffraction methods with in situ thermodiffractometry and thermal analyses to characterize the selective and reversible transformation of the [M(pmdc)(H(2)O)(2)] x H(2)O compounds (M = Fe, Co, Ni, Cu) into the bis-hydrated [M(pmdc)(H(2)O)(2)] counterparts by moderate heating, which is followed by an irreversible transformation into two-dimensional (2-D) anhydrous species. The structural features of the transient bis-hydrated species and of the completely dehydrated one are described for M = Cu. Remarkably, the first dehydration process does not alter the 1-D nature of the [M(pmdc)(H(2)O)(2)] chains; on the contrary, the second dehydration gives rise to the loss of the axially coordinated water molecules with a concomitant condensation of the 1-D chains into 2-D layers through ancillary carboxylate bridging groups. The magnetic properties of the anhydrous [M(pmdc)] species (M = Co, Ni, Cu) have been investigated, showing that these phases behave as 1-D antiferromagnets with interchain interactions. Notably, in the case of the [Ni(pmdc)] system, a weak ferromagnetic ordering, arising from a spin canting phenomenon with a blocking temperature of 13 K, is observed. PMID- 19281182 TI - Theoretical studies on metal-metal interaction, excited states, and spectroscopic properties of binuclear Au-Au, Au-Rh, and Rh-Rh complexes with diphosphine ligands: buildup of complexity from monomers to dimers. AB - To understand their photocatalytic activity and application in luminescent materials, a series of gold and rhodium phosphine complexes (mononuclear [Au(I)(PH(3))(2)](+) (1) and [Rh(I)(CNH)(2)(PH(3))(2)](+) (2); homobinuclear [Au(I)(2)(PH(2)CH(2)PH(2))(2)](2+) (3) and [Rh(I)(2)(CNH)(4)(PH(2)CH(2)PH(2))(2)](2+) (4); heterobinuclear [Au(I)Rh(I)(CNH)(2)(PH(2)CH(2)PH(2))(2)](2+) (5), [Au(I)Rh(I)(CNH)(2)(PH(2)NHPH(2))(2)Cl(2)] (6), and [Au(I)Rh(I)(CNH)(2)(PH(2)NHPH(2))(2)](2+) (7); and oxidized derivatives [Au(II)Rh(II)(CNH)(2)(PH(2)CH(2)PH(2))(2)](4+) (8), [Au(II)Rh(II)(CNH)(2)(PH(2)NHPH(2))(2)Cl(3)](+) (9), and [Au(II)Rh(II)(CNH)(2)(PH(2)NHPH(2))(2)](4+) (10)) were investigated using ab initio methods and density functional theory. With the use of the MP2 method, the M-M' distances in 3-7 were estimated to be in the range of 2.76-3.02 A, implying the existence of weak metal-metal interaction. This is further evident in the stretching frequencies and bond orders of M-M'. The two-electron oxidation from 5 7 to their respective partners 8-10 was shown to mainly occur in the gold-rhodium centers. Experimental absorption spectra were well reproduced by our time dependent density functional theory calculations. The metal-metal interaction results in a large shift of d(z(2)) --> p(z) transition absorptions in binuclear complexes relative to mononuclear analogues and concomitantly produces a low lying excited state that is responsible for increasing visible-light photocatalytic activities. Upon excitation, the metal-centered transition and the metal-to-metal charge transfer strengthen the metal-metal interaction in triplet excited states for 3-6, while the promotion of electrons into the sigma*(d(z(2))) orbital weakens the interaction in 9. PMID- 19281183 TI - Functional identification of incorrectly annotated prolidases from the amidohydrolase superfamily of enzymes. AB - The substrate profiles for two proteins from Caulobacter crescentus CB15 (Cc2672 and Cc3125) and one protein (Sgx9359b) derived from a DNA sequence ( gi|44368820 ) isolated from the Sargasso Sea were determined using combinatorial libraries of dipeptides and N-acyl derivatives of amino acids. These proteins are members of the amidohydrolase superfamily and are currently misannotated in NCBI as catalyzing the hydrolysis of l-Xaa-l-Pro dipeptides. Cc2672 was shown to catalyze the hydrolysis of l-Xaa-l-Arg/Lys dipeptides and the N-acetyl and N-formyl derivatives of lysine and arginine. This enzyme will also hydrolyze longer peptides that terminate in either lysine or arginine. The N-methyl phosphonate derivative of l-lysine was a potent competitive inhibitor of Cc2672 with a K(i) value of 120 nM. Cc3125 was shown to catalyze the hydrolysis of l-Xaa-l-Arg/Lys dipeptides but will not hydrolyze tripeptides or the N-formyl and N-acetyl derivatives of lysine or arginine. The substrate profile for Sgx9359b is similar to that of Cc2672 except that compounds with a C-terminal lysine are not recognized as substrates. The X-ray structure of Sgx9359b was determined to a resolution of 2.3 A. The protein folds as a (beta/alpha)(8)-barrel and self associates to form a homooctamer. The active site is composed of a binuclear metal center similar to that found in phosphotriesterase and dihydroorotase. In one crystal form, arginine was bound adventitiously to the eight active sites within the octamer. The orientation of the arginine in the active site identified the structural determinants for recognition of the alpha-carboxylate and the positively charged side chains of arginine-containing substrates. This information was used to identify 18 other bacterial sequences that possess identical or similar substrate profiles. PMID- 19281184 TI - Self-assembling peptide amphiphile-based nanofiber gel for bioresponsive cisplatin delivery. AB - The aim of this study is to develop a bioresponsive cisplatin (CDDP) delivery system with a self-assembling peptide amphiphile (PA) comprising a cell-adhesive matrix metalloproteinase-2 (MMP-2)-sensitive GTAGLIGQRGDS and a fatty acid. A biomimetic CDDP-PA gel was spontaneously formed upon incubating a mixture of CDDP and the PA for 5 h at 37 degrees C. CDDP-PA gel formation was confirmed by rheological analysis. The structure of self-assembled CDDP-PA nanofibers inside the gel was determined by transmission electron microscopy (TEM). Bioresponsive drug release from the biomimetic gel was demonstrated by in vitro MMP-2-triggered CDDP release. The MMP-2-sensitive CDDP release was dependent on the enzyme concentration in the medium. Enzymatic degradation of the CDDP-PA gel was confirmed by TEM images of the gel degraded in an MMP-2 containing medium. The MMP-2-triggered CDDP release as well as the presentation of RGDS in the gel would potentially provide a spatially and temporally controlled delivery system for targeted anticancer drug delivery. PMID- 19281185 TI - Nanoparticle processing in the solid state dramatically increases the cell membrane permeation of a cholesterol-lowering drug, probucol. AB - High cholesterol levels (or hypercholesterolemia) are linked with many diseases, particularly with the risk of coronary heart diseases. Probucol is commonly used to reduce cholesterol in blood. While the effectiveness of this drug highly depends on its solubility, unfortunately, it is nearly insoluble (solubility is 5 ng/mL in water). Therefore, it is essential to develop approaches to increase its solubility and bioavailability and to enhance the efficiency of the drug. Here we show that a new method increases the solubility of probucol in water and its ability to permeate cell membranes. This new method of processing the drug in a nanoparticle utilizes the grinding of PBC probucol together with sodium dodecylsulfate and methacrylic copolymer. Solid-state NMR experiments reveal the polymorphic state of probucol and the conversion of this drug from crystalline to the amorphous state, and determine its nearness to the copolymer due to the grinding process that enables the formation of nanoparticles. PMID- 19281186 TI - Alignment-free prediction of a drug-target complex network based on parameters of drug connectivity and protein sequence of receptors. AB - There are many drugs described with very different affinity to a large number of receptors. In this work, we selected drug-receptor pairs (DRPs) of affinity/nonaffinity drugs to similar/dissimilar receptors and we represented them as a large network, which may be used to identify drugs that can act on a receptor. Computational chemistry prediction of the biological activity based on quantitative structure-activity relationships (QSAR) substantially increases the potentialities of this kind of networks avoiding time- and resource-consuming experiments. Unfortunately, most QSAR models are unspecific or predict activity against only one receptor. To solve this problem, we developed here a multitarget QSAR (mt-QSAR) classification model. Overall model classification accuracy was 72.25% (1390/1924 compounds) in training, 72.28% (459/635) in cross-validation. Outputs of this mt-QSAR model were used as inputs to construct a network. The observed network has 1735 nodes (DRPs), 1754 edges or pairs of DRPs with similar drug-target affinity (sPDRPs), and low coverage density d = 0.12%. The predicted network has 1735 DRPs, 1857 sPDRPs, and also low coverage density d = 0.12%. After an edge-to-edge comparison (chi-square = 9420.3; p < 0.005), we have demonstrated that the predicted network is significantly similar to the one observed and both have a distribution closer to exponential than to normal. PMID- 19281187 TI - Cryogenic neon matrix-isolation FTIR spectroscopy of evaporated ionic liquids: geometrical structure of cation-anion 1:1 pair in the gas phase. AB - Low-temperature infrared spectra of thermally evaporated ionic liquids, 1-ethyl- and 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide and bis(trifluoromethanesulfonyl)amide have been measured in a cryogenic Ne matrix. The experimental IR spectrum of bis(trifluoromethanesulfonyl)amide can be reproduced theoretically by not B3LYP/6-31G* but MP2/6-31G* calculation, which suggests that the vibrational analysis for ionic liquids composed of bis(trifluoromethanesulfonyl)imide anion would be more successfully performed using the MP2 calculation. By comparison of the matrix-isolation spectra of the ionic liquids with the MP2 calculation, their geometrical structures in the gas phase are determined to be of C(2-position)-H(+)...N(-) interaction structure, which corresponds to the geometry of the energetically second-lowest ion-pair structure. The present study may provide a valuable clue to understand a vaporization mechanism of ionic liquid. PMID- 19281188 TI - Affinity of drugs and small biologically active molecules to carbon nanotubes: a pharmacodynamics and nanotoxicity factor? AB - The MM-PBSA MD method was used to estimate the affinity, as represented by log k(b), of each of a variety of biologically active molecules to a carbon nanotube in an aqueous environment. These ligand-receptor binding simulations were calibrated by first estimating the log k(b) values for eight ligands to human serum albumin, HSA, whose log k(b) values have been observed. A validation linear correlation equation was established [R(2) = 0.888, Q(2) = 0.603] between the observed and estimated log k(b) values to HSA. This correlation equation was then used to rescale all MM-PBSA MD log k(b) values using a carbon nanotube as the receptor. The log k(b) of the eight HSA ligands, nine polar and/or rigid ligands and six nonpolar and/or flexible ligands to a carbon nanotube were estimated. The range in rescaled log k(b) values across this set of 23 ligands is 0.25 to 7.14, essentially 7 orders of magnitude. Some ligands, like PGI2, bind in the log k(b) = 7 range which corresponds to the lower limits of known drugs. Thus, such significant levels of binding of biologically relevant compounds to carbon nanotubes might lead to alterations in the normal pharmacodynamic profiles of these compounds and be a source of toxicity. Ligand binding potency to a carbon nanotube is largely controlled by the shape, polarity/nonpolarity distribution and flexibility of the ligand. HSA ligands exhibit the most limited binding to a carbon nanotube, and they are relatively rigid and of generally spherical shape. Polar and/or rigid ligands bind less strongly to the carbon nanotube, on average, than nonpolar and/or flexible ligands even though the chosen members of both classes of ligands in this study have chainlike shapes that facilitate binding. The introduction of only a few strategically spaced single bonds in the polar and/or rigid ligands markedly increases their binding to a carbon nanotube. PMID- 19281189 TI - Biofunctionalized magnetic nanoparticle integrated mid-infrared pathogen sensor for food matrixes. AB - Magnetic nanoparticles functionalized with anti-Escherichia coli O157:H7 or anti Salmonella typhimurium antibodies that can specifically bind to their target organisms were used to isolate E. coli O157:H7 and S. typhimurium separately from a cocktail of bacteria and from food matrixes. The pathogens were then detected using label-free IR fingerprinting. The binding and detection protocol was first validated using a benchtop FT-IR spectrometer and then applied to a portable mid IR spectrometer to enable this approach as a point-of-detection technology. Highly selective detection was achieved in less than 30 min at both species (E. coli O157:H7 vs S. typhimurium ) and strain (E. coli O157:H7 vs E. coli K12) levels in complex food matrixes (2% milk, spinach extract) with a detection limit of 10(4)-10(5) CFU/mL. The combined approach of functionalized magnetic nanoparticles and IR spectroscopy imparts specificity through spectroscopic fingerprinting and selectivity through species-specific antibodies with an in built sample extraction step and could be applied in the field for on-site food borne pathogen monitoring. PMID- 19281190 TI - Handheld miniature ion trap mass spectrometers. AB - For field applications, "miniature" and "rapid" have become almost synonymous, yet these small mass spectrometers are not useful if performance is too severely compromised. (To listen to a podcast about this feature, please go to the Analytical Chemistry website at pubs.acs.org/journal/ancham .). PMID- 19281192 TI - Cationic liposomes target sites of acute neuroinflammation in experimental autoimmune encephalomyelitis. AB - The binding selectivity of charged liposomes to the spinal cord of rats affected by experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis, was investigated. Positively and negatively charged liposomes were injected into the tail vein of rats, and blood/brain barrier (BBB) targeting was determined by confocal microscopy as a function of the temporal evolution of the inflammatory response. Accumulation in spinal cord endoneural vessels was observed for cationic, but not for anionic, liposomes, and only in EAE but not in healthy rats. The overall binding efficacy paralleled the severity of the clinical score, but targeting was observed already before clinical manifestation of inflammation. Preferential binding of positively charged liposomes in the course of acute EAE can be ascribed to subtle changes of BBB morphology and charge distribution in a similar way as for the binding of cationic particles to proliferating vasculature in chronic inflammation and angiogenesis. Our findings suggest that vascular changes related to increased binding affinity for cationic particles are very early events within the inflammatory reaction in acute EAE. Investigation of cationic vascular targeting can help to shed further light on these occurrences, and, potentially, new diagnostic and therapeutic options may become available. In neuroinflammatory diseases, cationic colloidal carrier particles may enable intervention at affected BBB by an approach which is independent from permeability increase. PMID- 19281193 TI - Crystal and molecular structure of manganese(II) lapacholate, a novel polymeric species undergoing temperature-reversible metal to ligand electron transfer. AB - Lapachol (2-hydroxy-3-(3-methyl-2-butenyl)-1,4-naphtoquinone) (HLap, C(15)H(14)O(3)) reacts with Mn(2+) producing a novel polymeric complex with formula: [Mn(Lap)(2)](n). Two ligands chelate the metal through their ortho oxygen (O1, O2) moiety while two para oxygens, from other Lap ligands, complete the octahedral coordination sphere. Thus far, all reported Lap metal complexes are mononuclear, lack the metal-trans-quinonic (para) oxygen binding and have Lap as a bidentate ligand. Synthesis, X-ray diffraction, IR, and UV-visible spectroscopic properties, thermogravimetric analysis, and differential thermal analysis of this complex are reported along with a density functional theory study describing electron transfer from the Mn to the Lap ligand at low temperature. X-ray structure determinations at 125, 197, and 300 K describe the progressive trend of a Mn contribution to the Mn-O1 bond length as a function of T. The Mn-O1 bond distance increases with temperature and may be therefore associated with a semiquinonate action at low T by the carbonyl O1 donor (and corresponding to Mn(III)). It transforms to a more classical coordinative bond at room T and stabilizes a Mn(II) species; this is a reversible phenomenon involving Mn(II)-Mn(III) oxidation states. PMID- 19281194 TI - Kinetics of bromine-magnesium exchange reactions in substituted bromobenzenes. AB - Competition experiments have been performed to determine the relative reactivities of substituted bromobenzenes, bromonaphthalenes, and 9 bromoanthracene toward i-PrMgCl x LiCl in THF at 0 degrees C. The rates of the bromine-magnesium exchange reactions are accelerated by electron-acceptor substituents, the activating efficiency of which increases in the order para < meta << ortho. The activation free enthalpies of the bromine-magnesium exchange reactions correlate fairly (r(2) = 0.83) with the proton affinities of analogously substituted aryllithiums (slope 0.8). The kinetics of two representative bromoarenes with i-PrMgCl x LiCl were found to be first-order in both bromoarene and i-PrMgCl x LiCl. Combination of the resulting second-order rate constants with the k(rel) values from competition experiments allowed us to calculate reaction times for the bromine-magnesium exchange reactions of a large variety of bromoarenes. PMID- 19281195 TI - Synthesis of super stable triangulenium dye. AB - The super stabilized tris(dialkylamino)-azadioxatriangulenium carbenium ion (A ADOTA(+)) 4 has been synthesized and characterized. As a result of extended delocalization of the positive charge in the planar triangulenium system and the presence of several strongly electron-donating groups, the A-ATOTA(+) ion displays extreme cation stability with a pK(R+) value close to 25. Furthermore the A-ATOTA(+) ion shows excellent dye properties with intense absorption at 458 nm and high fluorescence quantum yield. PMID- 19281196 TI - High-pressure hetero-Diels-Alder route to (+/-)-6,6,6-trifluoro-beta-C-naphthyl glycosides. AB - The first de novo synthesis of a beta-C-naphthyl glycoside displaying a convenient functionality for subsequent transformations into complex C-aryl glycosides is reported. The synthesis of this (+/-)-beta-C-1,5 dibenzyloxynaphthyl 6,6,6-trifluoro-3-amino glycoside relies on a hyperbaric HDA reaction involving a new 2-vinylnaphthalenic dienophile. PMID- 19281197 TI - Structural study of Ga(III), In(III), and Fe(III) complexes of triaza-macrocycle based ligands with N3S3 donor set. AB - Two new ligands, 1,4,7-tris(2-mercaptoethyl)-1,4,7-triazacyclodecane (L2) and 9 methyl-1,4,7-tris(2-mercaptoethyl)-1,4,7-triazacyclodecane (L3), have been synthesized by reaction of the free triazacycloalkanes with ethylene sulfide. Complexes of L2 with Ga(III), In(III), and Fe(III), as well as Ga(III) and In(III) complexes of L3, have been characterized by single crystal X-ray diffraction. In all cases, the metal ions are coordinated in a trigonal antiprismatic environment with varying degrees of distortion. L2 complexes are isostructural and exhibit the chair-Delta(lambda lambda)(delta delta delta) conformation, whereas GaL3 and InL3 adopt twist-boat-Delta(lambda lambda)(delta delta delta) and twist-boat-Delta(lambda lambda)(lambda delta delta) configurational isomers and their enantiomers, respectively. In addition, we report the crystal structure of the Fe(III) complex of the known ligand 1,4,7 tris(2-mercaptoethyl)-1,4,7-triazacyclononane (TS-TACN, L1), which is isostructural to its Ga(III) and In(III) complexes. All aforementioned complexes are compared (including literature data of Ga(TS-TACN) and In(TS-TACN)), and the influence of ring size and backbone substitution on coordination geometries is discussed. Furthermore, the solution structures of Ga(III) and In(III) complexes of the new ligands have been investigated employing NMR spectroscopy and density functional theory (DFT) calculation. The latter revealed that for all compounds, the most stable structures are those where the six-membered chelate ring adopts chair conformation. NMR measurements confirmed this proposal for the L2 complexes. In contrast, the GaL3 complex is present as a mixture of chair and twist-boat isomers. This implies that for this complex, interconversion between both forms is occurring, which necessarily includes intermediate partial decoordination of the metal ion. InL3 occurs as a single species, but an unambiguous determination of its conformation was not possible. PMID- 19281198 TI - The SAMP1 solvation challenge: further lessons regarding the pitfalls of parametrization. AB - This paper describes an investigation into the performance of different charges and radii with the Poisson-Boltzmann method for the calculation of vacuum-water transfer energies. The test set consisted of 63 drug-like molecules used in a blind-test challenge. The results are consistent with earlier reports, namely, that more physical charges perform better and that radii parametrization can both improve and also dramatically worsen results, with the latter suggesting a failure to capture all of the basic physics of solvation. PMID- 19281199 TI - Concerning the reactivity of PTAD with isomeric dienes: the mechanism of the Diels-Alder cycloaddition. AB - Cyclopropyl substituted dienes are employed as mechanistic probes in the triazolinedione Diels-Alder (DA) reaction. In aprotic and protic solvents, apart from the DA adducts that bear an intact cyclopropyl group, complicated and rearranged products are also obtained. These results provide solid evidence for the involvement of an open intermediate with a lifetime greater than 2 x 10(-12) s. PMID- 19281200 TI - Efficient approach to determine the pK(a) of the proton release complex in the photocycle of retinal proteins. AB - This work utilizes a photoelectrochemical approach to study the pH dependence of proton release and uptake in the photocycles of two retinal proteins, bacteriorhodopsin (BR) and archaerhodopsin 4 (AR4). By detecting photoinduced potentials that originate from the proton concentration changes (DeltaH(+)) generated by proteins near the indium tin oxide (ITO) electrode, we show that the kinetics of release and uptake can be followed in a broad pH range, and the pK(a) of the proton release complex (PRC) can be easily determined under different conditions. Nonoriented protein films were deposited on the electrode, and photovoltage in an electrochemical cell was detected after illumination with a green flash. The kinetics of proton release and uptake could be measured as light induced decreases and increases of the photopotential. A kinetic analysis was performed, and a formula describing proton fluxes of wild-type BR and AR4 and D96N mutant of BR was derived. Three componentsfast proton release, slow proton release, and proton uptakewere found in the wild-type retinal proteins; two components, fast and slow proton releases, were found in the D96N mutant. The pH dependence of the fraction of fast release over the whole release was used to determine the pK(a) for proton release in the photocycles of these retinal proteins. Measurements were also performed in conventional buffer solutions and crown ether. The presence of buffer in 10-50 mM concentration did not abolish the light-induced signals, indicating that the electrode response is much less sensitive to buffers than pH-sensitive dyes in a suspension due to a higher protein/buffer ratio near the electrode. This feature enables us to study effects of chemicals with high buffer capacity, and significant effects of buffers and crown ether on proton pumping behaviors of retinal proteins were revealed. In comparison with the classic pH-sensitive dye approach, the photoelectrochemical approach is convenient and efficient for measurements of transient proton concentration changes (DeltaH(+)) generated by a proton pump and thus might be utilized as a powerful tool for the investigation of light-driven proton pumping mechanisms in a wide pH range. PMID- 19281201 TI - Raman spectroscopic study, DFT calculations and MD simulations on the conformational isomerism of N-alkyl-N-methylpyrrolidinium bis (trifluoromethanesulfonyl) amide ionic liquids. AB - The conformational behaviors of N-alkyl-N-methylpyrrolidinium bis (trifluoromethanesulfonyl) amide ionic liquids (alkyl; propyl and butyl, [P(1n)][TFSA]; n = 3 and 4) were studied by Raman spectroscopy in the frequency range of 200-1700 cm(-1) at different temperatures. Observed Raman spectra in the frequency range 870-960 cm(-1) for [P(13)][TFSA] and at 860-950 cm(-1) for [P(14)][TFSA] depend on the temperature, indicating that pseudo rotational isomerization of the pyrrolidinium ring exists in the ionic liquids. DFT calculations revealed that the pseudo rotational potential energy surfaces for P(13)(+) and P(14)(+) ions were similar to each other, i.e., the e6 isomer is the global minimum, whereas the three other isomers e1, e4, and e5 are ca. 3 kJ mol( 1) higher in energy. Optimized geometries with no imaginary frequency were successfully obtained for the e6, e1, and e4 isomers. For both cations, the theoretical Raman spectra of the e6 isomers reproduce well the observed data. To explain their observed Raman spectra in a reasonable way, it is necessary to consider one or more species as predicted by DFT calculations, i.e., the e4 isomer of P(13)(+) rather than the e1, or the e1 isomer of P(14)(+) rather than the e4. In addition, the torsion energy potentials of the alkyl chains of the cations were scanned by DFT calculations. It turns out that the alkyl chains of the cations prefer all trans conformations. It should be emphasized that the alkyl chains of the pyrrolidinium cations show remarkably different conformational behaviors comparing with those of the imidazolium. The isomerization enthalpies Delta(iso)H degrees from the e6 to the e4 isomer of P(13)(+) and to e1 of P(14)(+) were reasonably estimated from the temperature dependence of Raman spectra based on our proposed assignments to be 2.9 kJ mol( 1) for P(13)(+) and 4.2 kJ mol(-1) for P(14)(+), respectively. Thus evaluated experimental Delta(iso)H degrees values, which may contain some uncertainties, are in agreement with those predicted by DFT calculations and MD simulations suggesting that pseudo rotational isomerization equilibria are established in the examined N-alkyl-N-methylpyrrolidinium ionic liquids. The conformational behavior of TFSA(-) was also investigated. The Delta(iso)H degrees from the trans (trifluoromethyl groups on opposite sides of the S-N-S plane) to the cis isomer were evaluated to be 4.2 kJ mol(-1) for [P(13)][TFSA] and 3.5 kJ mol(-1) for [P(14)][TFSA], respectively, which are similar to that for the 1-ethyl 3methylimidazolium ionic liquid. PMID- 19281202 TI - Effect of aluminum on the local structure of silicon in zeolites as studied by Si K edge X-ray absorption near-edge fine structure: spectra simulation with a non muffin tin atomic background. AB - Experimental Si K edge X-ray absorption near-edge fine structure (XANES) of zeolite faujasite, mordenite, and beta are interpreted by means of the FEFF8 code, replacing the theoretical atomic background mu(0) by a background that was extracted from an experimental spectrum. To some extent, this diminished the effect of the inaccuracy introduced by the MT potential and accounted for the intrinsic loss of photoelectrons. The agreement of the theoretical and experimental spectra at energies above the white lines enabled us to identify structural distortion around silicon, which occurs with increasing aluminum content. The Si K edge XANES spectra are very sensitive to slight distortions in the silicon coordination. Placing an aluminum atom on a nearest neighboring T site causes a distortion in the silicon tetrahedron, shortening one of the silicon-oxygen bonds relative to the other three. PMID- 19281203 TI - Synthesis, reactivity, and kinetics of substitution in W3PdSe4 cuboidal clusters. A reexamination of the kinetics of substitution of the related W3S4 cluster with thiocyanate. AB - The reaction of Pd(dba)(2) (dba = dibenzylideneacetone) with [W(3)Se(4)(H(2)O)(9)](4+) in 2 M HCl gives the cuboidal cluster [W(3)(PdCl)Se(4)(H(2)O)(9)](3+), which undergoes edge-to-edge condensation and crystallizes from Hpts solutions as edge-linked double-cubane cluster [{W(3)PdSe(4)(H(2)O)(9)}(2)](pts)(8) x 18 H(2)O (pts(-) = p-toluenesulfonate). The substitution of Cl(-) by different ligands, including phenylsulfinate PhSO(2)(-), was explored. The phenylsulfinate complex was crystallized as a 2:1 adduct with cucurbit[6]uril (C(36)H(36)N(24)O(12)), [W(3)(Pd(PhSO(2))Se(4)(H(2)O)(8.58)Cl(0.42)](2)(C(36)H(36)N(24)O(12))Cl(5.16) x 16.83 H(2)O, and its structure was determined by X-ray diffraction. Solution studies indicate that the Pd atom is able to stabilize the pyramidal tautomer of hypophosphorous and phosphorous acid: HP(OH)(2) and P(OH)(3). Kinetic studies were carried out on the reactions with H(3)PO(2) and thiocyanate, which were found to proceed in two and three kinetically resolvable steps, respectively. The kinetic results are discussed in terms of the mechanistic proposals put forward in the literature for related complexes. To gain insight into the details of the substitution kinetics in these kinds of clusters, the reaction of the related [W(3)S(4)(H(2)O)(9)](4+) complex with NCS(-) has been reexamined, and the results obtained provide for the first time information about the rates of substitution of the whole set of nine-coordinated water molecules. PMID- 19281204 TI - Time-resolved resonance Raman and density functional theory investigation of the photoreactions of benzophenone in aqueous solution. AB - A time-resolved resonance Raman study of the photoreactions of benzophenone (BP) in neutral, alkaline, and acidic aqueous solutions is reported. Under neutral and alkaline conditions, a hydrogen abstraction reaction is observed to form the corresponding diphenyl ketyl (DPK) radical with different reaction rates while under acidic condition the protonation of triplet benzophenone occurs simultaneously, which is followed by a faster hydration reaction to produce the corresponding photohydration short-lived intermediates that were observed during experiments and that were tentatively assigned to the m-(3)BP x H(2)O and the o (1)BP x H(2)O species. The dynamics, structures, and properties of these species are briefly discussed. PMID- 19281205 TI - Palladium N(CH2CH2P(i)Pr2)2-dialkylamides: synthesis, structural characterization, and reactivity. AB - Palladium(II) aminodiphosphine PNP pincer complexes [PdR(PNP(H))]PF(6) (1(R); R = Cl Me, Ph; PNP(H) = HN(CH(2)CH(2)P(i)Pr(2))(2)) were prepared. Deprotonation with KO(t)Bu affords dialkylamides [PdR(PNP)] (2(R); R = Cl Me, Ph; PNP = (NCH(2)CH(2)P(i)Pr(2))(2)) in high yield which are stable toward beta-H elimination. While AgPF(6) oxidizes the amides, cationic amido complexes [PdL(PNP)]PF(6) (3(L); L = CN(t)Bu, PMe(3)) were obtained upon chloride abstraction from 1(Cl) with TlPF(6). The reaction of amide 2(Cl) with MeOTf results in N-methylation yielding [PdCl(PNP(Me))]OTf (5) quantitatively. N-H acidities of the amino complexes 1(Me) (pK(a) = 24.2(1)) and 1(Ph) (pK(a) = 23.2(1)) were determined in dmso. Complexes 1(Cl), 1(Me), 2(Cl) 2(Me), 3(CNtBu), and 5 were structurally characterized by single crystal X-ray diffraction. The amido complexes feature pyramidal nitrogen atoms in the solid state. The molecular structures, high N-basicity, and reactivity of the amido complexes can be explained with Pd-N(amido) bonding that is characterized by strong N-->Pd sigma-donation and repulsive d(pi)-p(pi) pi-interactions. This interpretation was confirmed by density functional theory (DFT) calculations of 2(Cl). PMID- 19281206 TI - Hydrogen bonds between zwitterions: intermediate between classical and charge assisted ones. A case study. AB - Detailed investigation of the electron density distribution function of N succinopyridine in the crystal and in isolated state within Bader's Atoms in Molecules theory has been carried out to analyze the charge density and energetical aspects of zwitterionic H-bonding. By means of comparative analysis of H-bonds formed between molecular, zwitterionic, and likely charged species, it was shown that, according to the criteria of H-bonding, all these interactions do not differ from each other. PMID- 19281207 TI - Experimental and theoretical study of the Br...N halogen bond in complexes of 1,4 dibromotetrafluorobenzene with dipyridyl derivatives. AB - The electron density distributions of two halogen-bonded complexes, that is, (E) 1,2-bis(4-pyridyl)ethylene (bpe) or 4,4'-dipyridyl (dp) with 1,4 dibromotetrafluorobenzene (C(6)F(4)Br(2)), have been obtained from accurate single-crystal X-ray diffracted intensities collected at 90 K and analyzed through the Bader's quantum theory of atoms in molecules. The experimental results have been compared with theoretical densities resulting from DFT calculations on both gas-phase isolated complexes and periodic crystal structures. The topological features and the energetics of the underlying Br...N intermolecular halogen bonding connecting bpe and dp with C(6)F(4)Br(2) molecules into 1D infinite chains have been investigated and compared with the previously analyzed I...N halogen bond. The analysis provides a quantitative evaluation of the differences observed between the involved halogen species, in addition to pointing out the basic features shared by the investigated halogen bond interactions. PMID- 19281208 TI - Cationic clathrate I Si(46-x)P(x)Te(y) (6.6(1) < or = y < or = 7.5(1), x < or = 2y): crystal structure, homogeneity range, and physical properties. AB - A new cationic clathrate I Si(46-x)P(x)Te(y) (6.6(1) < or = y < or = 7.5(1), x < or = 2y at 1375 K) was synthesized from the elements and characterized by X-ray powder diffraction, thermal analysis, scanning electron microscopy, wavelength dispersive X-ray spectroscopy (WDXS), neutron powder diffraction, and (31)P NMR spectroscopy. The thermal behaviors of the magnetic susceptibility and resistivity were investigated as well. Si(46-x)P(x)Te(y) reveals a wide homogeneity range due to the presence of vacancies in the tellurium guest positions inside the smaller cage of the clathrate I structure. The vacancy ordering in the structure of Si(46-x)P(x)Te(y) causes the change of space group from Pm3n (ideal clathrate I) to Pm3 accompanied by the redistribution of P and Si atoms over different framework positions. Neutron powder diffraction confirmed that P atoms preferably form a cage around the vacancy-containing tellurium guest position. Additionally, (31)P NMR spin-spin relaxation experiments revealed the presence of sites with different coordination of phosphorus atoms. Precise determination of the composition of Si(46-x)P(x)Te(y) by WDXS showed slight but noticeable deviation (x < or = 2y) of phosphorus content from the Zintl counting scheme (x = 2y). The compound is diamagnetic while resistivity measurements show activated behavior or that of heavily doped semiconductors. Thermal analysis revealed high stability of the investigated clathrate: Si(46-x)P(x)Te(y) melts incongruently at approximately 1460 K in vacuum and is stable in air against oxidation up to 1295 K. PMID- 19281209 TI - Infrared spectra and density functional theory calculations of group 10 transition metal sulfide molecules and complexes. AB - Laser-ablated Ni, Pd, and Pt atoms were reacted with sulfur molecules emerging from a microwave discharge in argon during condensation at 7 K. Reaction products were identified from matrix infrared spectra, sulfur isotopic shifts, spectra of sulfur isotopic mixtures, and frequencies from density functional calculations. The strongest absorptions are observed at 597.9, 596.1, and 583.6 cm(-1), respectively, for the group 10 metals. These absorptions show large sulfur-34 shifts and 32/34 isotopic frequency ratios (1.0282, 1.0285, 1.0298) that are appropriate for S-S stretching modes. Of most importance, mixed 32/34 isotopic 1/4/4/2/4/1 sextets identify this product with two equivalent S(2) molecules containing equivalent atomic positions as the bisdisulfur pi complexes M(S(2))(2). Our DFT calculations find stable D(2h) structures with B(1u) ground states and intense b(1u) infrared active modes a few wavenumbers higher than the observed values. A minor Ni product at 505.8, 502.7 cm(-1) shows the proper sulfur-34 shift for assignment to (58)NiS, (60)NiS. Another major product with Pt at 512.2 cm(-1) reveals an asymmetric triplet absorption with mixed sulfur 32/34, which is appropriate for assignment to the SPtS disulfide molecule. A weak 491.7 cm(-1) peak exhibits the sulfur-34 shift expected for PtS, and this assignment follows. PMID- 19281210 TI - Where is electronic energy stored in adenosine triphosphate? AB - The gas-phase electronic energy of the hydrolysis of methyl triphosphate, a model of adenosine 5'-triphosphate (ATP), is partitioned into local (atomic and group) contributions. A modified definition of Lipmann's "group transfer potential" is proposed on the basis of the partitioning of the total electronic energy into atomic contributions within the framework of the quantum theory of atoms in molecules (QTAIM). The group transfer potential is defined here as the sum of the atomic energies forming the group in ATP minus the sum of the energies of the same atoms in inorganic phosphate. It is found that the transfer potential of the terminal phosphate group in ATP is significantly reduced, from +241.7 to +73.1 kcal/mol, as a result of complexation with magnesium. This is accompanied by a concomitant change in the energy of reaction from -168.6 to -24.9 kcal/mol. Regions within ATP where the electronic energy changes the most upon hydrolysis are identified. The study is conducted at the DFT/B3LYP/6-31+G(d,p) level of theory. PMID- 19281211 TI - Strong conductance variation in conformationally constrained oligosilane tunnel junctions. AB - The effects of molecular conformation on conductance in oligosilane-bridged metal molecule-metal junctions are studied theoretically using density functional theory combined with a nonequilibrium Green's function approach. Varying the internal SiSiSiSi dihedral angles in hexasilane diamine chains changes the conductance by up to 3 orders of magnitude. This conformational dependence is due to the effects of sigma-delocalization on the positions of the highest occupied molecular orbital (HOMO) energies. The conductance values for the different conformations are related to electron transfer rates in donor-bridge-acceptor systems, and the effect of shifting the injection energy is examined. The transport properties are found to be extremely sensitive to the alignment between the HOMO energies and Fermi level of the gold electrodes. PMID- 19281212 TI - B2C graphene, nanotubes, and nanoribbons. AB - We report a first-principles prediction of a new two-dimensional inorganic material, namely, the B(2)C graphene in which the boron and carbon atoms are packed into a mosaic of hexagons and rhombuses. In the B(2)C graphene, each carbon atom is bonded with four boron atoms, forming a planar-tetracoordinate carbon (ptC) moiety, a notion first conceived by Hoffmann et al. The B(2)C graphene is possibly a metal with a small overlap in the energy of conduction and valence bands. Like the carbon graphene and nanotubes, a B(2)C graphene sheet can be rolled into various forms of B(2)C nanotubes as well. Depending on the roll-up vector, the B(2)C nanotubes may become either a metal or a semiconductor. All B(2)C graphene nanoribbons are predicted to be uniformly metallic, regardless of their width and edge structure. PMID- 19281213 TI - Noncovalent interactions of Ni+ with N-donor ligands (pyridine, 4,4'-dipyridyl, 2,2'-dipyridyl, and 1,10-phenanthroline): collision-induced dissociation and theoretical studies. AB - Kinetic-energy-dependent collision-induced dissociation (CID) of complexes of a variety of N-donor ligands (N-L) with Ni(+), Ni(+)(N-L)(x), is studied using guided ion beam tandem mass spectrometry. The N-donor ligands investigated include: pyridine, 4,4'-dipyridyl, 2,2'-dipyridyl, and 1,10-phenanthroline. For most of the Ni(+)(N-L)(x) complexes, CID results in endothermic loss of a single neutral N-L ligand as the primary dissociation pathway. Sequential dissociation of additional N-L ligands is observed at elevated energies for the pyridine and 4,4'-dipyridyl complexes containing more than one ligand. The cross-section thresholds for the primary dissociation pathways are interpreted to yield 0 and 298 K bond dissociation energies (BDEs) of the Ni(+)(N-L)(x) complexes after accounting for the effects of multiple ion-neutral collisions, the kinetic and internal energy distributions of the reactants, and their lifetimes for dissociation. Density functional theory calculations at the B3LYP/6 311+G(2d,2p)//B3LYP/6-31G* level are performed to obtain model structures, molecular parameters, and energetics for the neutral N-L ligands and the Ni(+)(N L)(x) complexes. In general, theory is found to overestimate the strength of binding to the first N-L ligand, and underestimate the strength of binding to additional ligands. Trends in the sequential BDEs of the Ni(+)(N-L)(x) complexes are examined and compared to complexes of Ni(+), to several other ligands previously investigated. The trends in the sequential BDEs are primarily determined by the valence electronic configuration and the effects of sd hybridization of Ni(+) but are also influenced by repulsive ligand-ligand interactions. Natural bond orbital analyses indicate that the binding in these complexes is primarily noncovalent. PMID- 19281214 TI - Electron tomography on micrometer-thick specimens with nanometer resolution. AB - Transmission electron microscopy (TEM) is a well-established technique to explore matter down to the atomic scale. TEM tomography methods have been developed to obtain volume information at the mesoscopic dimensions of devices or complex mixtures of multiphase objects with nanometer resolution, but these methods are in general only applicable to relatively thin specimens with a few hundred nanometer thickness at most. Here we introduce an approach based on scanning TEM (STEM) tomography that pushes the resolution in three dimensions down to a few nanometers for several micrometer ultrathick specimens using a conventional TEM with 300 kV accelerating voltage, and we demonstrate its versatility for materials research and nanotechnology. PMID- 19281215 TI - Adaptive logic circuits with doping-free ambipolar carbon nanotube transistors. AB - A CMOS-like inverter was integrated by using ambipolar carbon nanotube (CNT) transistors without doping. The ambipolar CNT transistors automatically configure themselves to play a role as an n-type or p-type transistor in a logic circuit depending on the supply voltage (V(DD)) and ground. A NOR (NAND) gate is adaptively converted to a NAND (NOR) gate. This adaptiveness of logic gates exhibiting two logic gate functions in a single logic circuit offers a new opportunity for designing logic circuits with high integration density for next generation applications. PMID- 19281216 TI - Sensing mechanisms for carbon nanotube based NH3 gas detection. AB - There has been an argument on carbon nanotube (CNT) based gas detectors with a field-effect transistor (FET) geometry: do the response signals result from charge transfer between adsorbed gas molecules and the CNT channel and/or from the gas species induced Schottky barrier modulation at the CNT/metal contacts? To differentiate the sensing mechanisms, we employed three CNTFET structures, i.e., (1) the entire CNT channel and CNT/electrode contacts are accessible to NH(3) gas; (2) the CNT/electrode contacts are passivated with a Si(3)N(4) thin film, leaving the CNT channel open to the gas and, in contrast, (3) the CNT channel is covered with the film, while the contacts are open to the gas. We suggest that the Schottky barrier modulation at the contacts is the dominant mechanism from room temperature to 150 degrees C. At higher temperatures, the charge transfer process contributes to the response signals. There is a clear evidence that the adsorption of NH(3) on the CNT channel is facilitated by environmental oxygen. PMID- 19281217 TI - Highly efficient vertical growth of wall-number-selected, N-doped carbon nanotube arrays. AB - We demonstrate a straightforward approach for rapid growth of wall-number selected, N-doped CNT arrays. Highly uniform nanopatterned iron catalyst arrays were prepared by tilted deposition through block copolymer nanotemplates. PECVD growth of CNTs from the nanopatterned catalysts in an NH(3) environment generated vertical N-doped CNTs with a fine-tunability of their carbon wall numbers. The optimized growth conditions produced 52 microm long N-doped CNTs within 1 min. Owing to N-doping, the wall-number selected CNTs including DWNTs and TWNTs demonstrated enhanced electro-conductivity and chemical functionality. This remarkably fast growth of highly uniform N-doped CNTs, whose material properties and chemical functionalizability are reinforced by N-doping, offers a new area of large-scale nanofabrication, potentially useful for diverse nanodevices. PMID- 19281218 TI - Functional characterization of ttmM unveils new tautomycin analogs and insight into tautomycin biosynthesis and activity. AB - The biosynthetic gene cluster for tautomycin (TTM), a potent protein phosphatase (PP) inhibitor has recently been characterized. Inactivation of ttmM, which encodes a putative C3' hydroxylase, afforded mutant SB6005 which accumulated three new 3'-deshydroxy TTM analogs, supporting the function of TtmM and the previously proposed linear pathway for TTM biosynthesis. Bioassays reveal the importance of the C3' OH moiety in PP inhibition and that PP inhibition is not the exclusive mechanism driving TTM-induced cell death. PMID- 19281219 TI - Enantioselective total synthesis of (-)-pironetin: iterative aldol reactions of thiazolidinethiones. AB - The enantioselective total synthesis of pironetin has been achieved in 11 steps from known aldehyde 2. The synthesis relies on the formation of 5 out of 6 stereocenters through titanium mediated iterative aldol reactions. Key steps in this synthesis include an acetal aldol reaction to establish the stereochemistry at C8 and C9, an acetate aldol reaction, and "Evans" syn aldol reaction. PMID- 19281220 TI - Imidazolylsulfonates: electrophilic partners in cross-coupling reactions. AB - Aryl imidazolylsulfonates participate as electrophilic coupling partners in palladium-mediated cross-coupling reactions. The aryl imidazolylsulfonates display good stability while maintaining good reactivity in a variety of palladium-catalyzed coupling reactions. Imidazolylsulfonates are a practical and economic alternative to triflates. PMID- 19281221 TI - In vitro skin diffusion study of pure forskolin versus a forskolin-containing Plectranthus barbatus root extract. AB - An in vitro skin diffusion study of pure forskolin (1) versus a 1-containing Plectranthus barbatus root extract (P. barbatus extract) in hairless guinea pig skin and human skin in a flow-through diffusion cell system was conducted and is being reported for the first time. Both topical agents were formulated in a solution of 70% ethanol and 30% propylene glycol (v/v). The results showed that forskolin can be delivered through the stratum corneum and that the flux of this compound was enhanced when 1 was delivered as a constituent of the P. barbatus extract as compared to an equivalent amount in pure form. These results suggest that the P. barbatus extract used contains permeation enhancement activity from other compound(s) contained in the crude root extract. It is possible that P. barbatus root extract may be used as an economical source of 1 to perform topical chemical manipulation of pigmentation in high-risk populations. PMID- 19281222 TI - Promiscuous aggregate-based inhibitors promote enzyme unfolding. AB - One of the leading sources of false positives in early drug discovery is the formation of organic small molecule aggregates, which inhibit enzymes nonspecifically at micromolar concentrations in aqueous solution. The molecular basis for this widespread problem remains hazy. To investigate the mechanism of inhibition at a molecular level, we determined changes in solvent accessibility that occur when an enzyme binds to an aggregate using hydrogen-deuterium exchange mass spectrometry. For AmpC beta-lactamase, binding to aggregates of the small molecule rottlerin increased the deuterium exchange of all 10 reproducibly detectable peptides, which covered 41% of the sequence of beta-lactamase. This suggested a global increase in proton accessibility upon aggregate binding, consistent with denaturation. We then investigated whether enzyme-aggregate complexes were more susceptible to proteolysis than uninhibited enzyme. For five aggregators, trypsin degradation of beta-lactamase increased substantially when beta-lactamase was inhibited by aggregates, whereas uninhibited enzyme was generally stable to digestion. Combined, these results suggest that the mechanism of action of aggregate-based inhibitors proceeds via partial protein unfolding when bound to an aggregate particle. PMID- 19281223 TI - Synthesis and antibacterial activities of N-glycosylated derivatives of tyrocidine A, a macrocyclic peptide antibiotic. AB - An efficient and practical method for macrocyclic glycopeptide synthesis was developed and utilized to synthesize tyrocidine A and its glycosylated derivatives. The method is based on solid-phase peptide synthesis using 2 chlorotrityl resin as the solid-phase support and glycosyl amino acids as building blocks. After glycopeptides with fully protected glycans and side chains were released from the acid-labile resin, their C- and N-termini were intramolecularly coupled in solution to afford cyclic glycopeptides in quantitative yields. This synthetic method should be generally applicable to various macrocyclic glycopeptides. Biological studies of the synthetic tyrocidine A derivatives showed that linking glycans directly to the Asn residue of tyrocidine A diminished its antibacterial activity, but linking glycans to Asn via a simple spacer did not. These results revealed the important impact of glycans on the activities, and probably the structures, of glycopeptide antibiotics. PMID- 19281224 TI - Electrochemical atomic-force microscopy using a tip-attached redox mediator. Proof-of-concept and perspectives for functional probing of nanosystems. AB - This paper presents the first steps toward the development of a new type of high resolution AFM-SECM microscopy which relies on the use of tip-attached redox labeled polymer chains as mediators to probe the local electrochemical reactivity of a planar substrate at the nanoscale. Submicrometer-sized combined gold AFM SECM probes were functionalized by linear, nanometer-sized, flexible, PEG3400 chains bearing a ferrocene (Fc) redox label at their free end. Analysis of the force and current approach curves recorded when such Fc-PEGylated probes (tips) were approached to a bare gold substrate allowed the presence of the Fc-PEG chains at the very tip end of the combined probes to be specifically demonstrated. It also allowed the chain coverage, configuration, and dynamics to be determined. When the Fc-PEGylated probe is positioned some approximately 5 nm above the substrate, only a few hundred chains are actually electrochemically contacting the surface, thus reducing the size of the tip-substrate interaction area to 20-40 nm. Most importantly, we have shown that the tip-borne PEG chains are flexible enough to allow their Fc heads to efficiently "sense" locally the electrochemical reactivity of the substrate, thus validating the working principle of the new AFM-SECM microscopy we propose. This innovative microscopy, we label Tarm (for tip-attached redox mediator)/AFM-SECM, should be particularly suitable for probing the activity of slowly functioning nanometer-sized active sites on surfaces, such as individual enzyme molecules, because it is, by design, free of the diffusional constraints which hamper the characterization of such nanosystems by classical SECM. PMID- 19281225 TI - Indolylarylsulfones bearing natural and unnatural amino acids. Discovery of potent inhibitors of HIV-1 non-nucleoside wild type and resistant mutant strains reverse transcriptase and coxsackie B4 virus. AB - New potent indolylarylsulfone (IAS) HIV-1 NNRTIs were obtained by coupling natural and unnatural amino acids to the 2-carboxamide and introducing different electron-withdrawing substituents at position 4 and 5 of the indole nucleus. The new IASs inhibited the HIV-1 replication in human T-lymphocyte (CEM) cells at low/subnanomolar concentration and were weakly cytostatic. Against the mutant L100I, K103N, and Y181C RT HIV-1 strains in CEM cells, sulfones 3, 4, 19, 27, and 31 were comparable to EFV. The new IASs were inhibitors to Coxsackie B4 virus at low micromolar (2-9 microM) concentrations. Superimposition of PLANTS docked conformations of IASs 19 and 9 revealed different hydrophobic interactions of the 3,5-dimethylphenyl group, for which a staking interaction with Tyr181 aromatic side chain was observed. The binding mode of 19 was not affected by the L100I mutation and was consistent with the interactions reported for the WT strain. PMID- 19281226 TI - Lipophilic lysine-spermine conjugates are potent polyamine transport inhibitors for use in combination with a polyamine biosynthesis inhibitor. AB - Cancer cells can overcome the ability of polyamine biosynthesis inhibitors to completely deplete their internal polyamines by the importation of polyamines from external sources. This paper discusses the development of a group of lipophilic polyamine analogues that potently inhibit the cellular polyamine uptake system and greatly increase the effectiveness of polyamine depletion when used in combination with DFMO, a well-studied polyamine biosynthesis inhibitor. The attachment of a length-optimized C(16) lipophilic substituent to the epsilon nitrogen atom of an earlier lead compound, D-Lys-Spm (5), has produced an analogue, D-Lys(C(16)acyl)-Spm (11) with several orders of magnitude more potent cell growth inhibition on a variety of cultured cancer cell types including breast (MDA-MB-231), prostate (PC-3), melanoma (A375), and ovarian (SK-OV-3), among others. These results are discussed in the context of a possible membrane catalyzed interaction with the extracellular polyamine transport apparatus. The resulting novel two-drug combination therapy targeting cellular polyamine metabolism has shown exceptional efficacy against cutaneous squamous cell carcinomas (SCC) in a transgenic ornithine decarboxylase (ODC) mouse model of skin cancer. A majority (88%) of large, aggressive SCCs exhibited complete or nearly complete remission to this combination therapy, whereas responses to each agent alone were poor. The availability of a potent polyamine transport inhibitor allows, for the first time, for a real test of the hypothesis that starving cells of polyamines will lead to objective clinical response. PMID- 19281227 TI - Dissecting molecular mechanisms in the living brain of dementia patients. AB - Understanding the molecular mechanisms associated with the development of dementia is essential for designing successful interventions. Dementia, like cancer and cardiovascular disease, requires early detection to potentially arrest or prevent further disease progression. By the time a neurologist begins to manage clinical symptoms, the disease has often damaged the brain significantly. Because successful treatment is the logical goal, detecting the disease when brain damage is still limited is of the essence. The role of chemistry in this discovery process is critical. With the advent of molecular imaging, the understanding of molecular mechanisms in human neurodegenerative diseases has exploded. Traditionally, knowledge of enzyme and neurotransmitter function in humans has been extrapolated from animal studies, but now we can acquire data directly from both healthy and diseased human subjects. In this Account, we describe the use of molecular imaging probes to elucidate the biochemical and cellular bases of dementia (e.g., Alzheimer's disease) and the application of these discoveries to the design of successful therapeutic interventions. Molecular imaging permits observation and evaluation of the basic molecular mechanisms of disease progression in the living brains of patients. 2-Deoxy-2 [(18)F]fluoro-d-glucose is used to assess the effect of Alzheimer's disease progression on neuronal circuits projecting from and to the temporal lobe (one of the earliest metabolic signs of the disease). Recently, we have developed imaging probes for detection of amyloid neuropathology (both tau and beta-amyloid peptide deposits) and neuronal losses. These probes allow us to visualize the development of pathology in the living brain of dementia patients and its consequences, such as losses of critical neurons associated with memory deficits and other neuropsychiatric impairments. Because inflammatory processes are tightly connected to the brain degenerative processes, inflammation is now emerging as an important target for new molecular imaging probes. The combination of molecular probes targeting various processes of dementia is a useful tool for detailed monitoring of disease mechanism, progression, and diagnosis, as well as for the development of rational strategies for promising therapeutic interventions. PMID- 19281229 TI - Helical wrapping of single-walled carbon nanotubes by water soluble poly(p phenyleneethynylene). AB - Amphiphilic, linear conjugated poly[p-{2,5-bis(3 propoxysulfonicacidsodiumsalt)}phenylene]ethynylene (PPES) efficiently disperses single-walled carbon nanotubes (SWNTs) under ultrasonication conditions into the aqueous phase. Vis-NIR absorption spectroscopy, atomic force microscopy (AFM), and transmission electron microscopy (TEM) demonstrate that these solubilized SWNTs are highly individualized. AFM and TEM data reveal that the interaction of PPES with SWNTs gives rise to a self-assembled superstructure in which a polymer monolayer helically wraps the nanotube surface; the observed PPES pitch length (13 +/- 2 nm) confirms structural predictions made via molecular dynamics simulations. This work underscores design elements important for engineering well defined nanotube-semiconducting polymer hybrid structures. PMID- 19281230 TI - Alterations in actin cytoskeletal assembly and junctional protein complexes in human endothelial cells induced by dengue virus infection and mimicry of leukocyte transendothelial migration. AB - Vascular leakage is a hallmark of severe dengue infection. Although extensive studies have been conducted during the past several decades, the molecular mechanisms underlying vascular leakage in dengue shock syndrome (DSS) remain unclear. We thus performed a proteomics study to characterize responses in human endothelial cells (EA.hy926) after DEN-2 virus infection (MOI=10). Comparative 2 D PAGE analysis revealed significantly altered abundance levels of 15 proteins, which were successfully identified by quadrupole time-of-flight mass spectrometry (MS) and/or tandem MS (MS/MS). These altered proteins were involved in several biological processes, for example, mRNA stability/processing, transcription and translation regulation, molecular chaperoning, oxidative stress response/regulation, cytoskeletal assembly, protein degradation, and cellular metabolisms. We also performed functional analyses of alterations in actin cytoskeletal assembly and endothelial integrity focusing on adherens junction (VE cadherin), tight junction (ZO-1) and adhesive molecule (PECAM-1) after 24-h of DEN-2 infection and simulation of transendothelial migration by PECAM-1 cross linking. Decreased expression and disorganization of the actin-cytoskeleton were observed in the infected cells, whereas the increase in actin stress fibers was found in adjacent noninfected cells. Additionally, a decrease in adhesive protein PECAM-1 was observed. Furthermore, DEN-2 infection caused decreased expression and redistribution of both VE-cadherin and ZO-1, whose changes were enhanced by PECAM-1 engagement. These alterations may potentially be a molecular basis explaining increased endothelial permeability or vascular leakage in DSS. PMID- 19281231 TI - Acyl transfer catalysis with 1,2,4-triazole anion. AB - 1,2,4-Triazole anion has been identified as an active acyl transfer catalyst suitable for the aminolysis and transesterification of esters. PMID- 19281232 TI - Characterization of salt bridges to lysines in the protein G B1 domain. AB - NMR investigations have been carried out on the B1 domain of protein G. This protein has six lysine residues, of which three are consistently found to form surface-exposed salt bridges in crystal structures, while the other three are not. The Nzeta and Hzeta chemical shifts of all six lysines are similar and are not affected significantly by pH titration of the carboxylate groups in the protein, except for a relatively small titration of K39 Nzeta. Deuterium isotope effects on nitrogen and proton are of the size expected for a simple hydrated amine (a result supported by density functional theory calculations), and also do not titrate with the carboxylates. The line shapes of the J-coupled (15)N signals suggest rapid internal reorientation of all NH(3)(+) groups. pK(a) values have been measured for all charged side chains except Glu50 and do not show the perturbations expected for salt bridge formation, except that E35 has a Hill coefficient of 0.84. The main differential effect seen is that the lysines that are involved in salt bridges in the crystal display faster exchange of the amine protons with the solvent, an effect attributed to general base catalysis by the carboxylates. This explanation is supported by varying buffer composition, which demonstrates reduced electrostatic shielding at low concentration. In conclusion, the study demonstrates that the six surface-exposed lysines in protein G are not involved in significant salt bridge interactions, even though such interactions are found consistently in crystal structures. However, the intrahelical E35-K39 (i,i+4) interaction is partially present. PMID- 19281233 TI - Phase transformation of biphasic Cu2S-CuInS2 to monophasic CuInS2 nanorods. AB - We synthesized wurtzite CuInS(2) nanorods (NRs) by colloidal solution-phase growth. We discovered that the growth process starts with nucleation of Cu(2)S nanodisks, followed by epitaxial overgrowth of CuInS(2) NRs onto only one face of Cu(2)S nanodisks, resulting in biphasic Cu(2)S-CISu heterostructured NRs. The phase transformation of biphasic Cu(2)S-CuInS(2) into monophasic CuInS(2) NRs occurred with growth progression. The observed epitaxial overgrowth and phase transformation is facile for three reasons. First, the sharing of the sulfur sublattice by the hexagonal chalcocite Cu(2)S and wurtzite CuInS(2) minimizes the lattice distortion. Second, Cu(2)S is in a superionic conducting state at the growth temperature of 250 degrees C wherein the copper ions move fluidly. Third, the size of the Cu(2)S nanodisks is small, resulting in fast phase transformation. Our results provide valuable insight into the controlled solution growth of ternary chalcogenide nanoparticles and will aid in the development of solar cells using ternary I-III-VI(2) semiconductors. PMID- 19281234 TI - Zinc incorporation via the vapor-liquid-solid mechanism into InP nanowires. AB - We report the incorporation of zinc atoms into vapor-liquid-solid grown indium phosphide nanowires via a gold catalyst particle. We demonstrate this by synthesizing axial pn-junctions, chemically etching them, and fabricating electrical contacts in a vertical configuration. Electrical measurements show clear diode behavior. Control of dopant incorporation is crucial for future applications and will eventually lead to full freedom of design. PMID- 19281235 TI - Hairpin structure of a biarsenical-tetracysteine motif determined by NMR spectroscopy. AB - The biarsenical-tetracysteine motif is a useful tag for genetic labeling of proteins with small molecules in living cells. The present study concerns the structure of a 12 amino acid peptide FLNCCPGCCMEP bound to the fluorophore ReAsH based on resorufin. (1)H NMR spectroscopy was used to determine the solution structure of the complex formed between the peptide and the ReAsH moiety. Structure calculations based on the NMR results showed that the backbone structure of the peptide is fairly well defined, with a hairpinlike turn, similar to a type-II beta-turn, formed by the central CPGC segment. The most stable complex was formed when As2 was bonded to C4 and C5 and As1 to C8 and C9. Two clear NOESY cross-peaks between the Phe1 side chain and ReAsH confirmed the close positioning of the phenyl ring of Phe1 and ReAsH. Phe1 was found to have an edge face geometry relative to ReAsH. The close interaction between Phe1 and ReAsH may be highly significant for the fluorescence properties of the ReAsH complex. PMID- 19281236 TI - Oleylamine-mediated synthesis of Pd nanoparticles for catalytic formic acid oxidation. AB - We report a facile synthesis of monodisperse Pd nanoparticles by the reduction of Pd(acac)(2) with oleylamine and borane tributylamine complex. The oleylamine coated Pd nanoparticles are readily "cleaned" with a 99% acetic acid wash, and the Pd particles supported on Ketjen carbon are catalytically active for formic acid oxidation in HClO(4) solution. The catalyst shows no obvious activity degradation after 1500 cyclic voltammetry cycles under ambient conditions. These Pd particles hold promise as a highly active non-Pt catalyst for fuel cell applications. PMID- 19281237 TI - Using a hydrazone-protected benzenediazonium salt to introduce a near-monolayer of benzaldehyde on glassy carbon surfaces. AB - A methodology is described for introducing a thin layer of covalently attached benzaldehyde on glassy carbon surfaces using aryl diazonium chemistry. Usually the electroreduction of aryl diazonium salts leads to the formation of an ill defined multilayer because of the involvement of highly reactive aryl radicals that can add to already-grafted aryl groups. However, in this study we used a two step "formation-degradation" procedure to solve this problem with the first step consisting of an electrografting of an aryl diazonium salt of a long-chain and bulky alkyl hydrazone onto a glassy carbon surface. The design of the hydrazone group serves to minimize multilayer formation by greatly diminishing the grafting rate after the first-layer formation and at the same time preventing radical additions from taking place at the inner aryl ring. Another valuable property of the hydrazone group is that it easily can be deprotected to the corresponding aldehyde by acid hydrolysis (i.e., the degradation step). In this manner, a thin and well-defined film of covalently attached benzaldehyde with an estimated coverage of 4 x 10(-10) mol cm(-2) was formed. The electrochemical responses of benzaldehyde were highly reproducible and largely independent of grafting medium (water or DMSO) and along with that also the thickness of the initially grafted film. AFM and contact angle measurements support the findings. The "formation degradation" approach thus lays the foundation for carrying out further functionalization reactions in a controlled manner. PMID- 19281238 TI - Effect of glucuronosylation on anthocyanin color stability. AB - The effect of glucuronosylation on the color stability of anthocyanins was investigated using glucuronosylated anthocyanins isolated from the flower petals of the red daisy (Bellis perennis) or obtained by enzymatic in vitro synthesis using heterologously expressed red daisy glucuronosyltransferase BpUGT94B1. Color stability toward light and heat stress was assessed by monitoring CIELAB color coordinates and stability at pH 7.0 by A(550). Cyanidin-3-O-2''-O glucuronosylglucoside showed improved color stability in response to light compared to both cyanidin 3-O-glucoside and cyanidin 3-O-2''-O-diglucoside. A similar increase in color stability was not observed following heat treatment. Glucuronosylation did not increase the stability of anthocyanins at pH 7.0 as determined by A(550). To test for a possible effect of glucuronosylation on the color stability of anthocyanins in plant extracts used for food coloration, an elderberry (Sambucus nigra) extract was glucuronosylated in vitro. Glucuronosylation of approximately 50% of total anthocyanins proceeded fast and resulted in increased color stability in response to both heat and light. The data show that glucuronosylation may be used to stabilize industrially used extracts of natural colorants. PMID- 19281239 TI - Direct asymmetric Michael addition to nitroalkenes: vinylogous nucleophilicity under dinuclear zinc catalysis. AB - Under dinuclear catalysis, the direct conjugate addition of 2(5H)-furanone to nitroalkenes involves the gamma-position of the nucleophile. The synthetically versatile Michael adducts are prepared in good yields, with high levels of diastereo- and enantioselectivity. A model is presented to rationalize the observed stereoselectivity. PMID- 19281240 TI - Cerebrospinal fluid proteomics reveals potential pathogenic changes in the brains of SIV-infected monkeys. AB - The HIV-1-associated neurocognitive disorder occurs in approximately one-third of infected individuals. It has persisted in the current era of antiretroviral therapy, and its study is complicated by the lack of biomarkers for this condition. Since the cerebrospinal fluid is the most proximal biofluid to the site of pathology, we studied the cerebrospinal fluid in a nonhuman primate model for HIV-1-associated neurocognitive disorder. Here we present a simple and efficient liquid chromatography-coupled mass spectrometry-based proteomics approach that utilizes small amounts of cerebrospinal fluid. First, we demonstrate the validity of the methodology using human cerebrospinal fluid. Next, using the simian immunodeficiency virus-infected monkey model, we show its efficacy in identifying proteins such as alpha-1-antitrypsin, complement C3, hemopexin, IgM heavy chain, and plasminogen, whose increased expression is linked to disease. Finally, we find that the increase in cerebrospinal fluid proteins is linked to increased expression of their genes in the brain parenchyma, revealing that the cerebrospinal fluid alterations identified reflect changes in the brain itself and not merely leakage of the blood-brain or blood-cerebrospinal fluid barriers. This study reveals new central nervous system alterations in lentivirus induced neurological disease, and this technique can be applied to other systems in which limited amounts of biofluids can be obtained. PMID- 19281241 TI - Listeria monocytogenes phosphatidylinositol-specific phospholipase C: Kinetic activation and homing in on different interfaces. AB - The phosphatidylinositol-specific phospholipase C (PI-PLC) from Listeria monocytogenes forms aggregates with anionic lipids leading to low activity. The specific activity of the enzyme can be enhanced by dilution of the protein or by addition of both zwitterionic and neutral amphiphiles (e.g., diheptanoylphosphatidylcholine or Triton X-100) or 0.1-0.2 M inorganic salts. Activation by amphiphiles occurs with both micellar (phosphatidylinositol dispersed in detergents) and monomeric [dibutroylphosphatidylinositol (diC(4)PI)] phosphotransferase substrates and inositol 1,2-(cyclic)-phosphate (cIP), the phosphodiesterase substrate. The presence of zwitterionic and neutral amphiphiles (to which the protein binds weakly) dilutes the surface concentration of the interfacial anionic substrate and thereby reduces the level of enzyme phospholipid particle aggregation. Zwitterionic amphiphiles also can bind directly to the protein and enhance catalysis since they enhance both diC(4)PI and cIP hydrolysis. In contrast to activation by amphiphiles, the rate enhancement by salt occurs for only the phosphotransferase step of the reaction. Added salt has a synergistic effect with zwitterionic phospholipids, leading to high specific activities for PI cleavage with only moderate dilution of the anionic substrate in the interface. This kinetic activation correlates with weakening of strong PI-PLC hydrophobic interactions with the interface as monitored by a decrease in the maximum monolayer surface pressure for insertion of the protein. Several point mutations of surface hydrophobic residues (W49A, L51A, L235A, and F237W) can dramatically alter the unusual kinetics of this secreted enzyme. The high affinity of PI-PLC for anionic phospholipids along with a strong hydrophobic interaction, which gives rise to the unusual kinetic behavior, is considered in terms of how it might contribute to the role of this phospholipase in L. monocytogenes infectivity. PMID- 19281242 TI - Anti-amyloid activity of the C-terminal domain of proSP-C against amyloid beta peptide and medin. AB - Amyloid fibrils are found in approximately 25 different diseases, including Alzheimer's disease. Lung surfactant protein C (SP-C) forms fibrils in association with pulmonary disease. It was recently found that the C-terminal domain of proSP-C (CTC), which is localized to the endoplasmic reticulum (ER) lumen, protects the transmembrane (TM) part of (pro)SP-C from aggregation into amyloid until it has a folded into an alpha-helix. CTC appears to have a more general anti-amyloid effect by also acting on TM regions of other proteins. Here we investigate interactions of CTC with the amyloid beta-peptide (Abeta) associated with Alzheimer's disease and medin, a peptide that forms fibrils in the most common form of human amyloid. CTC prevents fibril formation in Abeta and medin and forms a complex with Abeta oligomers, as judged by size-exclusion chromatography and electrospray ionization mass spectrometry. These data suggest that CTC functions as a chaperone that acts preferentially against unfolded TM segments and structural motifs found during amyloid fibril formation, a mechanism that may be exploited in forming a basis for future anti-amyloid therapy. PMID- 19281243 TI - Method revealing bacterial cell-wall architecture by time-dependent isotope labeling and quantitative liquid chromatography/mass spectrometry. AB - The molecular details of the biosynthesis and resulting architecture of the bacterial cell wall remain unclear but are essential to understanding the activity of glycopeptide antibiotics, the recognition of pathogens by hosts, and the processes of bacterial growth and division. Here we report a new strategy to elucidate bacterial cell-wall architecture based on time-dependent isotope labeling of bacterial cells quantified by liquid chromatography/accurate mass measurement mass spectrometry. The results allow us to track the fate of cell wall precursors (which contain the vancomycin-binding site) in Enterococcus faecium, a leading antibiotic-resistant pathogen. By comparing isotopic enrichments of postinsertionally modified cell-wall precursors, we find that tripeptides and species without aspartic acid/asparagine (Asp/Asn, Asx) bridges are specific to mature cell wall. Additionally, we find that the sequence of cell wall maturation varies throughout a cell cycle. We suggest that actively dividing E. faecium cells have three zones of unique peptidoglycan processing. Our results reveal new organizational characteristics of the bacterial cell wall that are important to understanding tertiary structure and designing novel drugs for antibiotic-resistant pathogens. PMID- 19281244 TI - Robust fluorescent detection of protein fatty-acylation with chemical reporters. AB - Fatty-acylation of proteins in eukaryotes is associated with many fundamental cellular processes but has been challenging to study due to limited tools for rapid and robust detection of protein fatty-acylation in cells. The development of azido-fatty acids enabled the nonradioactive detection of fatty-acylated proteins in mammalian cells using the Staudinger ligation and biotinylated phosphine reagents. However, the visualization of protein fatty-acylation with streptavidin blotting is highly variable and not ideal for robust detection of fatty-acylated proteins. Here we report the development of alkynyl-fatty acid chemical reporters and improved bioorthogonal labeling conditions using the Cu(I) catalyzed Huisgen [3 + 2] cycloaddition that enables specific and sensitive fluorescence detection of fatty-acylated proteins in mammalian cells. These improvements allow the rapid and robust biochemical analysis of fatty-acylated proteins expressed at endogenous levels in mammalian cells by in-gel fluorescence scanning. In addition, alkynyl-fatty acid chemical reporters enable the visualization of fatty-acylated proteins in cells by fluorescence microscopy and flow cytometry. The ability to rapidly visualize protein fatty-acylation in cells using fluorescence detection methods therefore provides new opportunities to interrogate the functions and regulatory mechanisms of fatty-acylated proteins in physiology and disease. PMID- 19281245 TI - Nanoscale imaging of surface topography and reactivity with the scanning electrochemical microscope. AB - Over the last 2 decades, scanning electrochemical microscopy (SECM) has been extensively employed for topographic imaging and mapping surface reactivity on the micrometer scale. We used flat, polished nanoelectrodes as SECM tips to carry out feedback mode imaging of various substrates with nanoscale resolution. Constant-height and constant-current images of plastic and Au compact disc surfaces and more complicated objects (computer chips and wafers) were obtained. The possibility of simultaneous imaging of surface topography and electrochemical reactivity was demonstrated. Very fast mass transfer at nanoelectrodes allowed us to obtain high-quality electrochemical images in viscous media under steady-state conditions, e.g., in 1-methyl-3-octylimidazolium bis(tetrafluoromethylsulfonyl)imide (C(8)mimC(1)C(1)N) ionic liquid. Ion-transfer based imaging was also performed using nanopipets as SECM tips. PMID- 19281246 TI - A crystalline imine-linked 3-D porous covalent organic framework. AB - A new crystalline porous three-dimensional covalent organic framework, termed COF 300, has been synthesized and structurally characterized. Tetrahedral tetra-(4 anilyl)-methane and linear terephthaldehyde building blocks were condensed to form imine linkages in a material whose X-ray crystal structure shows five independent diamond frameworks. Despite the interpenetration, the structure has pores of 7.2 A diameter. Thus, COF-300 shows thermal stability up to 490 degrees C and permanent porosity with a surface area of 1360 m(2) g(-1). PMID- 19281247 TI - New one-pot four-component synthesis of disubstituted pyrido[2,3-d]pyrimidine-6 carboxamide derivatives. AB - In this work, 1,2,3,4,5,8-hexahydro-1,3,7-trimethyl-2,4-dioxopyrido[2,3 d]pyrimidine-6-carboxamide derivatives were synthesized in a simple and efficient method from the four-component condensation reaction of diketene, an aliphatic or aromatic amine, an aromatic aldehyde, and 6-amino-1,3-dimethyluracil in the presence of a catalytic amount of p-toluenesulfonic acid under mild conditions at ambient temperature in high yields. PMID- 19281248 TI - A lysine-tyrosine pair carries out acid-base chemistry in the metal ion-dependent pyridine dinucleotide-linked beta-hydroxyacid oxidative decarboxylases. AB - This work reviews published structural and kinetic data on the pyridine nucleotide-linked beta-hydroxyacid oxidative decarboxylases. The family of metal ion-dependent pyridine nucleotide-linked beta-hydroxyacid oxidative decarboxylases can be divided into two structural families with the malic enzyme, which has an (S)-hydroxyacid substrate, comprising one subfamily and isocitrate dehydrogenase, isopropylmalate dehydrogenase, homoisocitrate dehydrogenase, and tartrate dehydrogenase, which have an (R)-hydroxyacid substrate, comprising the second subclass. Multiple-sequence alignment of the members of the (R) hydroxyacid family indicates a high degree of sequence identity with most of the active site residues conserved. The three-dimensional structures of the members of the (R)-hydroxyacid family with structures available superimpose on one another, and the active site structures of the enzymes have a similar overall geometry of residues in the substrate and metal ion binding sites. In addition, a number of residues in the malic enzyme active site are also conserved, and the arrangement of these residues has a similar geometry, although the (R) hydroxyacid and (S)-hydroxyacid family sites are geometrically mirror images of one another. The active sites of the (R)-hydroxyacid family have a higher positive charge density when compared to those of the (S)-hydroxyacid family, largely due to the number of arginine residues in the vicinity of the substrate alpha-carboxylate and one fewer carboxylate ligand to the divalent metal ion. Data available for all of the enzymes in the family have been considered, and a general mechanism that makes use of a lysine (general base)-tyrosine (general acid) pair is proposed. Differences exist in the mechanism for generating the neutral form of lysine so that it can act as a base. PMID- 19281249 TI - Prostate-specific antigen is a "chymotrypsin-like" serine protease with unique P1 substrate specificity. AB - Prostate-specific antigen (PSA), a serine protease belonging to the human kallikrein family, is best known as a prostate cancer biomarker. Emerging evidence suggests that PSA may also play a salient role in prostate cancer development and progression. With large amounts of enzymatically active PSA continuously and selectively produced by all stages of prostate cancer, PSA is an attractive target. PSA inhibitors, therefore, may represent a promising class of therapeutics and/or imaging agents. PSA displays chymotrypsin-like specificity, cleaving after hydrophobic residues, in addition to possessing a unique ability to cleave after glutamine in the P1 position. In this study, we investigated the structural motifs of the PSA S1 pocket that give it a distinct architecture and specificity when compared to the S1 pocket of chymotrypsin. Using the previously described PSA substrate Ser-Ser-Lys-Leu-Gln (SSKLQ) as a template, peptide aldehyde based inhibitors containing novel P1 aldehydes were made and tested against both proteases. Glutamine derivative aldehydes were highly specific for PSA while inhibitors with hydrophobic P1 aldehydes were potent inhibitors of both proteases with K(i) values <500 nM. The crystal structure of PSA was used to generate a model that allowed GOLD docking studies to be performed to further understand the critical interactions required for inhibitor binding to the S1 pockets of PSA and chymotrypsin. In conclusion, these results provide experimental and structural evidence that the S1 specificity pocket of PSA is distinctly different from that of chymotrypsin and that the development of highly specific PSA inhibitors is feasible. PMID- 19281250 TI - C-F bond-cleavage reactions of fluoroalkanes with magnesium reagents and without metal catalysts. AB - An unexpected C-F bond-cleavage reaction of unactivated fluoroalkanes with the well-known Grignard reagents without using metal catalysts has been discovered. For example, a reaction between 1-fluorooctane and phenyl magnesium chloride gave n-octylbenzene in moderate yield. This coupling reaction via the activation of an unactivated alkyl carbon-fluorine bond proceeds with phenylmagnesium chloride, whereas methylmagnesium chloride did not give the C-C cross-coupling product but rather a halogen exchange product. PMID- 19281251 TI - Highly enantioselective construction of axial chirality by palladium-catalyzed cycloisomerization of N-alkenyl arylethynylamides. AB - A cationic palladium(II)/(S)-xyl-Segphos complex catalyzes enantioselective cycloisomerizations of N-alkenyl arylethynylamides leading to axially chiral 4 aryl-2-pyridones in high yields with high ee values. The present catalysis represents the first enantioselective construction of axial chirality by the transition-metal-catalyzed cycloisomerization. PMID- 19281252 TI - Mass thickness analysis of gold thin films using room temperature gas-phase chlorination. AB - Chemical interaction of ultrathin gold films with gaseous chlorine at room temperature results in quantitative metal chlorination and formation of Au(III) chloride. Combination of gas-phase chlorination of Au nanostructures with dissolution of the chloride product and spectrophotometric determination of the chloroaurate concentration presents a simple procedure for quantitative determination of the mass thickness of Au nanostructures. The method is demonstrated by mass thickness analysis of the condensation coefficient of Au adatoms on solid substrates, the extinction coefficient of Au nanoparticles, as well as study of the integrity of Au island films upon interaction with solvents. PMID- 19281253 TI - Silicon nanowire-based solar cells on glass: synthesis, optical properties, and cell parameters. AB - Silicon nanowire (SiNW)-based solar cells on glass substrates have been fabricated by wet electroless chemical etching (using silver nitrate and hydrofluoric acid) of 2.7 microm multicrystalline p(+)nn(+) doped silicon layers thereby creating the nanowire structure. Low reflectance (<10%, at 300-800 nm) and a strong broadband optical absorption (>90% at 500 nm) have been measured. The highest open-circuit voltage (V(oc)) and short-circuit current density (J(sc)) for AM1.5 illumination were 450 mV and 40 mA/cm(2), respectively at a maximum power conversion efficiency of 4.4%. PMID- 19281254 TI - Fano resonances in individual coherent plasmonic nanocavities. AB - We observe the appearance of Fano resonances in the optical response of plasmonic nanocavities due to the coherent coupling between their superradiant and subradiant plasmon modes. Two reduced-symmetry nanostructures probed via confocal spectroscopy, a dolmen-style slab arrangement and a ring/disk dimer, clearly exhibit the strong polarization and geometry dependence expected for this behavior at the individual nanostructure level, confirmed by full-field electrodynamic analysis of each structure. In each case, multiple Fano resonances occur as structure size is increased. PMID- 19281255 TI - Self-assembled monolayers of metal-assembling dendron thiolate formed from dendrimers with a disulfide core. AB - Novel phenylazomethine dendrimers with a disulfide core (SS-DPA G1-4) were synthesized in nearly quantitative yields. Although the disulfide core is shielded by the rigid dendron shell, direct formation of the self-assembled monolayers of metal-assembling dendron thiolate was observed by XPS and electrochemical reduction of the self-assembled monolayer substrates. The dendrimers showed a similar metal-assembling manner with other derivatives. The metal assembly to the self-assembled monolayers of metal-assembling dendron thiolate was also confirmed. PMID- 19281256 TI - Energetics of C-H bonds formed at single-walled carbon nanotubes. AB - We have calculated the adsorption energies for different hydrogen dimers adsorbed at the surface of zigzag single-walled carbon nanotubes (SWCNs) (n,0) (for n = 6 28) to identify the range of nanotube structural parameters that provide optimal adsorption energetics. We determined that, for H(2) gas in equilibrium with adsorbed hydrogen dimers, under normal conditions, carbon nanotubes with diameters in the range of 0.77 +/- 0.19 nm have a minimum energy overhead for hydrogen release and uptake in the hydrogen storage process. Most interestingly, we also found that the adsorption energies of hydrogen are correlated to the modulations of the nanotube density of states, because of the quantum confinement of electrons along the circumference of the SWCN. This effect is discussed from the perspective of chemical bond formation and is related to the valence to conduction band excitation energy. PMID- 19281257 TI - Proton-transfer and H2-elimination reactions of trimethylamine alane: role of dihydrogen bonding and Lewis acid-base interactions. AB - Proton-transfer and H(2)-elimination reactions of aluminum hydride AlH(3)(NMe(3)) (TMAA) with XH acids were studied by means of IR and NMR spectroscopy and DFT calculations. The dihydrogen-bonded (DHB) intermediates in the interaction of the TMAA with XH acids (CH(3)OH, (i)PrOH, CF(3)CH(2)OH, adamantyl acetylene, indole, 2,3,4,5,6-pentafluoroaniline, and 2,3,5,6-tetrachloroaniline) were examined experimentally at low temperatures, and the spectroscopic characteristics, dihydrogen bond strength and structures, and the electronic and energetic characteristics of these complexes were determined by combining experimental and theoretical approaches. The possibility of two different types of DHB complexes with polydentate proton donors (typical monodentate and bidentate coordination with the formation of a symmetrical chelate structure) was shown by DFT calculations and was experimentally proven in solution. The DHB complexes are intermediates of proton-transfer and H(2)-elimination reactions. The extent of this reaction is very dependent on the acid strength and temperature. With temperature increases the elimination of H(2) was observed for OH and NH acids, yielding the reaction products with Al-O and Al-N bonds. The reaction mechanism was computationally studied. Besides the DHB pathway for proton transfer, another pathway starting from a Lewis complex was discovered. Preference for one of the pathways is related to the acid strength and the nucleophilicity of the proton donor. As a consequence of the dual Lewis acid-base nature of neutral aluminum hydride, participation of a second ROH molecule acting as a bifunctional catalyst forming a six-member cycle connecting aluminum and hydride sites notably reduces the reaction barrier. This mechanism could operate for proton transfer from weak OH acids to TMAA in the presence of an excess of proton donor. PMID- 19281258 TI - Physiologically relevant online electrochemical method for continuous and simultaneous monitoring of striatum glucose and lactate following global cerebral ischemia/reperfusion. AB - This study demonstrates a new electroanalytical method with a high physiological relevance for simultaneous online monitoring of glucose and lactate in the striatum of the rat brain following global cerebral ischemia/reperfusion. The online analytical method is based on the efficient integration of in vivo microdialysis sampling with an online selective electrochemical detection with the electrochemical biosensors with dehydrogenases, i.e., glucose and lactate dehydrogenases, as recognition elements. The dehydrogenase-based electrochemical biosensors are developed onto the dual split-disk plastic carbon film (SPCF) electrodes with methylene green (MG) adsorbed onto single-walled carbon nanotubes (SWNTs) as the electrocatalyst for the oxidation of dihydronicotiamide adenine dinucleotide (NADH) at a low potential of 0.0 V (vs Ag/AgCl). Artificial cerebrospinal fluid (aCSF) containing NAD(+) is externally perfused from a second pump and online mixed with the brain microdialysates to minimize the variation of pH that occurred following the cerebral ischemia/reperfusion and to supply NAD(+) cofactor and O(2) for the enzymatic reactions of dehydrogenases and ascorbate oxidase, respectively. As a result, the developed online electroanalytical method exhibits a high selectivity against the electrochemically active species endogenously existing in the cerebral systems and a high tolerance against the variation of pH and O(2) following cerebral ischemia/reperfusion. This property, along with the good linearity and a high stability toward glucose and lactate as well as little cross-talk between two biosensors, substantially makes this method possible for the continuous, simultaneous, and online monitoring of glucose and lactate in the rat brain following global cerebral ischemia/reperfusion. This study establishes a new and effective platform for the investigation of the energy metabolism in physiological and pathological processes. PMID- 19281259 TI - Simultaneous transmission mode collision-induced dissociation and ion/ion reactions for top-down protein identification/characterization using a quadrupole/time-of-flight tandem mass spectrometer. AB - Simultaneous transmission mode collision-induced dissociation (CID) and ion/ion proton transfer reactions have been implemented on a quadrupole/time-of-flight (TOF) tandem mass spectrometer. Reagent anions were trapped in a pressurized quadrupole collision cell by applying appropriate dc voltages while multiply protonated protein precursor ions were injected into the collision cell at energies sufficient to give rise to CID. Intact precursor ions as well as fragment ions underwent ion/ion proton transfer reactions during their passage through the collision cell and on to an orthogonal acceleration TOF mass analyzer. The resulting product ion spectrum was then submitted to deconvolution to yield a "zero-charge" spectrum, which was then matched against in silico produced spectra derived from a protein database. Dramatic improvements in the scores associated with correct matches were obtained relative to CID data without the benefit of ion/ion reactions for proteins as large as carbonic anhydrase (29 kDa). The parameters that most affect the extent of ion/ion proton transfer during transmission through the instrument include the number of anions stored in the collision cell, the amplitude of the radio frequency trapping voltage, the voltage of the LINAC potential associated with the collision cell, and the collision gas pressure. This work demonstrates that it is possible to effect whole protein tandem mass spectrometry with simultaneous CID, ion/ion reactions, and mass analysis for high duty cycle top-down protein characterization. PMID- 19281260 TI - Single conducting polymer nanowire chemiresistive label-free immunosensor for cancer biomarker. AB - A simple and cost-effective, all-electrochemical method to fabricate and assemble single conducting polymer nanowire based biosensors was developed. Polypyrrole (Ppy) nanowires were synthesized by electrochemical polymerization using an alumina template. The single-nanowire chemoresistive sensor device was assembled using ac dielectrophoretic alignment followed by maskless anchoring on a pair of gold electrodes separated by 3 microm. To establish an efficient covalent surface biofunctionalization route, glutaraldehyde (GA) and N-(3-dimethylaminopropyl)-N' ethylcarbodiimide hydrochloride (EDC) chemistries were compared. EDC was established to be the most effective chemistry and was used to surface functionalize a single Ppy nanowire with cancer antigen (CA 125) antibody to fabricate a nanoimmunosensor for CA 125 biomarker detection and quantification. The immunosensor had excellent sensitivity with a lower detection limit of 1 U/mL CA 125 and dynamic range up to 1000 U/mL in 10 mM phosphate buffer. Furthermore, there was no loss of performance upon exposure to CA 125 in spiked human blood plasma. This demonstrates the clinical importance of these sensors for cancer marker detection with cost benefits and great portability for diagnosis of patients at the point of care. PMID- 19281261 TI - Arrays of dual nanomechanical resonators for selective biological detection. AB - Arrays of small nanomechanical resonators with dual geometry have been fabricated for sensitive biological detection. The arrays consist of silicon nitride resonating 100 nm thick cantilevers with sensing gold areas alternately placed on the free and fixed cantilever ends. The Au areas act as sensing regions as can be functionalized by means of thiol chemistry. The nanomechanical arrays provide a double flavor of the adsorbed molecules: the added mass reported by the cantilevers with the Au area at the tip and the nanoscale elasticity reported by the cantilevers with the Au area at the clamp. The devices were applied for DNA detection based on Watson-Crick pairing rules. The proposed design for nanomechanical resonators provides higher specificity for DNA sensing in comparison with conventional single cantilevers. The nanoscale elasticity induced by the DNA hybridization arises from the intermolecular interactions between the adsorbates bound to the cantilever and the surface stress. PMID- 19281262 TI - Developing repeatable measurements for reliable analysis of molecules at surfaces using desorption electrospray ionization. AB - Desorption electrospray ionization (DESI) is a powerful ambient ionization technique that can provide high-sensitivity mass spectrometry information directly from surfaces at ambient pressure. Although a growing amount of research has been devoted to exploring different applications, there are few studies investigating the basic parameters and underpinning metrology. An understanding of these is crucial to develop DESI as the robust and reliable technique required for significant uptake by industry. In this work, we begin with a systematic study of the parameters affecting the repeatability, sensitivity, and rate of consumption of material with DESI. To do this we have developed a model sample consisting of a thin uniform film of controlled thickness of Rhodamine B on glass. This model sample allowed assessment of optimal sensitivity and spot shape under different conditions. In addition, it allowed us to study the surface in more detail to understand why and how each parameter affects these. Using the model sample to optimize the instrument parameters for DESI led to an absolute intensity repeatability of better than 15%, achieved over a period of 1 day. This model sample provides valuable insight into the electrospray-sample interaction and the desorption mechanism. Confocal microscopy of areas analyzed by DESI allow droplet distribution, material utilization, and spot size to be determined. Studying surface erosion also gives the erosion rate of material, analogous to the sputtering yield in secondary ion mass spectrometry. The results of the study provide a clear description that explains the differences observed with changing electrospray parameters allowing optimization of the technique, for both spatial resolution and sensitivity. PMID- 19281264 TI - Mimicking the structure of the V3 epitope bound to HIV-1 neutralizing antibodies. AB - The third variable region (V3) of the HIV-1 envelope glycoprotein gp120 is a target for virus neutralizing antibodies. The V3 sequence determines whether the virus will manifest R5 or X4 phenotypes and use the CCR5 or CXCR4 chemokine coreceptor, respectively. Previous NMR studies revealed that both R5- and X4-V3 peptides bound to antibodies 0.5beta and 447-52D form beta-hairpin conformations with the GPGR segment at the turn. In contrast, in their free form, linear V3 peptides and a cyclic peptide consisting of the entire 35-residue V3 loop were highly unstructured in aqueous solution. Herein we evaluated a series of synthetic disulfide constrained V3-peptides in which the position of the disulfide bonds, and therefore the ring size, was systematically varied. NMR structures determined for singly and doubly disulfide constrained V3-peptides in aqueous solution were compared with those found for unconstrained V3(JRFL) and V3(IIIB) peptides bound to 447-52D and to 0.5beta, respectively. Our study indicated that cyclic V3 peptides manifested significantly reduced conformational space compared to their linear homologues and that in all cases cyclic peptides exhibited cross-strand interactions suggestive of beta-hairpin-like structures. Nevertheless, the singly constrained V3-peptides retained significant flexibility and did not form an idealized beta-hairpin. Incorporation of a second disulfide bond results in significant overall rigidity, and in one case, a structure close to that of V3(MN) peptide bound to 447-52D Fab was assumed and in another case a structure close to that formed by the linear V3(IIIB) peptide bound to antibody 0.5beta was assumed. PMID- 19281265 TI - Why amantadine loses its function in influenza m2 mutants: MD simulations. AB - Molecular dynamics simulations of the drug-resistant M2 mutants, A30T, S31N, and L26I, were carried out to investigate the inhibition of M2 activity using amantadine (AMT). The closed and open channel conformations were examined via non and triply protonated H37. For the nonprotonated state, these mutants exhibited zero water density in the conducting region, and AMT was still bound to the channel pore. Thus, water transport is totally suppressed, similar to the wild type channel. In contrast, the triply protonated states of the mutants exhibited a different water density and AMT position. A30T and L26I both have a greater water density compared to the wild-type M2, while for the A30T system, AMT is no longer inside the pore. Hydrogen bonding between AMT and H37 crucial for the bioactivity is entirely lost in the open conformation. The elimination of this important interaction of these mutations is responsible for the lost of AMT's function in influenza A M2. This is different for the S31N mutant in which AMT was observed to locate at the pore opening region and bond with V27 instead of S31. PMID- 19281266 TI - Gene expression profiling of rat hippocampus following exposure to the acetylcholinesterase inhibitor soman. AB - Soman (O-pinacolyl methylphosphonofluoridate) is a potent neurotoxicant. Acute exposure to soman causes acetylcholinesterase inhibition, resulting in excessive levels of acetylcholine. Excessive acetylcholine levels cause convulsions, seizures, and respiratory distress. The initial cholinergic crisis can be overcome by rapid anticholinergic therapeutic intervention, resulting in increased survival. However, conventional treatments do not protect the brain from seizure-related damage, and thus, neurodegeneration of soman-sensitive brain areas is a potential postexposure outcome. We performed gene expression profiling of the rat hippocampus following soman exposure to gain greater insight into the molecular pathogenesis of soman-induced neurodegeneration. Male Sprague-Dawley rats were pretreated with the oxime HI-6 (l-(((4 aminocarbonyl)pyridinio)methoxyl)methyl)-2-((hydroxyimino)methyl)-pyridinium dichloride; 125 mg/kg, ip) 30 min prior to challenge with soman (180 microg/kg, sc). One minute after soman challenge, animals were treated with atropine methyl nitrate (2.0 mg/kg, im). Hippocampi were harvested 1, 3, 6, 12, 24, 48, 72, 96, and 168 h after soman exposure and RNA extracted to generate microarray probes for gene expression profiling. Principal component analysis of the microarray data revealed a progressive alteration in gene expression profiles beginning 1 h postexposure and continuing through 24 h postexposure. At 48 h to 168 h postexposure, the gene expression profiles clustered nearer to controls but did not completely return to control profiles. On the basis of the principal component analysis, analysis of variance was used to identify the genes most significantly changed as a result of soman at each postexposure time point. To gain insight into the biological relevance of these gene expression changes, genes were rank ordered by p-value and categorized using gene ontology-based algorithms into biological functions, canonical pathways, and gene networks significantly affected by soman. Numerous signaling and inflammatory pathways were identified as perturbed by soman. These data provide important insights into the molecular pathways involved in soman-induced neuropathology and a basis for generating hypotheses about the mechanism of soman-induced neurodegeneration. PMID- 19281267 TI - Diverse self-assembly in soluble oligoazaacenes: a microscopy study. AB - The synthesis and morphologies of self-assembled aggregates of novel oligoazapentacene 2 and oligoazaheptacene 3 derivatives are reported. Double nucleophilic substitution on 2,3-dicyano-[h,j]-dibenzo-1,4,5,10-tetrazaanthracene 4 gives the corresponding dihydro-oligoacene derivatives, which were then N alkylated using n-dodecyl bromide to yield self-assembling acene molecules. 2 and 3 self-assemble in solution, leading to a variety of aggregated structures including rolled-up sheets, foams, and fibrous structures reminiscent of organogels. These structures are of substantial interest because of their potential electronic properties and because individual fibers can be "exfoliated". Structures of the aggregates are discussed. Additionally, the crystal structure of precursor 4 is reported because it gives information regarding the intermolecular interactions (hydrogen bonding and intermolecular stacking) in similar compounds. Crystal data for 4: space group P2(1)/n, a = 9.3164(17) angstroms, b = 7.0649(13) angstroms, c = 23.684(4) angstroms, alpha = 90.00 degrees, beta = 99.945(3) degrees, gamma = 90.00 degrees, and V = 1535.4(5) angstroms3. PMID- 19281268 TI - Panorganismal gut microbiome-host metabolic crosstalk. AB - Coevolution shapes interorganismal crosstalk leading to profound and diverse cellular and metabolic changes as observed in gut dysbiosis in human diseases. Here, we modulated a simplified gut microbiota using pro-, pre-, and synbiotics to assess the depth of systemic metabolic exchanges in mice, using a multicompartmental modeling approach with metabolic signatures from 10 tissue/fluid compartments. The nutritionally induced microbial changes modulated host lipid, carbohydrate, and amino acid metabolism at a panorganismal scale. Galactosyl-oligosaccharides reduced lipogenesis, triacylglycerol incorporation into lipoproteins and triglyceride concentration in the liver and the kidney. Those changes were not correlated with decreased plasma lipoproteins that were specifically induced by L. rhamnosus supplementation. Additional alteration of transmethylation metabolic pathways (homocysteine-betaine) was observed in the liver and the pancreas following pre- and synbiotic microbial modulation, which may be of interest for control of glucose metabolism and insulin sensitivity. Probiotics also reduced hepatic glycogen and glutamine and adrenal ascorbate with inferred effects on energy homeostasis, antioxidation, and steroidogenesis. These studies show the breadth and the depth of gut microbiome modulations of host biochemistry and reveal that major mammalian metabolic processes are under symbiotic homeostatic control. PMID- 19281269 TI - Particle deposition onto charge-heterogeneous substrates. AB - The deposition of model colloidal particles onto striped charge-heterogeneous surfaces was studied to determine the influence of surface chemical heterogeneity on the deposit morphology. The charge heterogeneity was created employing self assembled monolayers of carboxyl- and amine-terminated alkanethiols using a soft lithographic technique. Polystyrene sulfate microspheres and fluorescent polystyrene nanoparticles were sequentially deposited onto the patterned substrate under no flow (quiescent) condition. The deposited structures and the micropatterns were imaged using a combination of phase contrast and fluorescence microscopy. The experimental particle deposition behavior was compared to predictions based on random sequential adsorption (RSA) employing a Monte Carlo technique. Comparison of radial distribution obtained from experimental data was made with the theoretical results and found to be in good agreement despite the use of a simple binary probabilistic model in the simulations. The primary conclusion from the study is that particles tend to preferentially deposit at the edges of the favorable stripes. However, the extent of this bias can be controlled by the proximity of consecutive favorable stripes (or width of the intervening unfavorable stripes) as well as the particle size relative to the stripe width. Second, a simple binary probability distribution-based Monte Carlo RSA deposition model adequately predicts the deposit structure, particularly the periodicity of the underlying patterns on the substrate. These observations suggest that the patterns could be encrypted by the deposited particles, which can subsequently be decoded, given the proper "key" or information that is based on analyzing the deposit morphology. PMID- 19281270 TI - 13C NMR spectroscopy of "Arduengo-type" carbenes and their derivatives. PMID- 19281272 TI - Chemical and biological characterization of thiol SAMs for neuronal cell attachment. AB - Cellular adhesion and growth on solid-state surfaces is the central theme in the development of cell-based biosensors and implantable medical devices. Suitable interface techniques must be applied to construct stable and well-organized thin films of biologically active molecules that would control the development of neuronal cells on chips. Peptides such as RGD fragments, poly-L-lysine (PLL), or basal lamina proteins, such as laminin or fibronectin, are often used in order to promote cellular adhesion on surfaces. In this paper we describe the characterization of several self-assembled monolayers (SAMs) for their ability to anchor a laminin-derived synthetic peptide, PA22-2, a peptide known to promote neuronal attachment and stimulate neurite outgrowth. We have evaluated the immobilization of PA22-2 onto 16-mercaptohexadecanoic acid, 4-maleimide-N-(11 undecyldithio)butanamide, and 2-(maleimide)ethyl-N-(11-hexaethylene oxide undecyldithio)acetamide SAM functionalized Au substrates. The neuronal attachment and outgrowth have been evaluated in embryonic mouse hippocampal neuron cultures up to 14 days in vitro. Our results show that differences in the cell morphologies were observed on the surfaces modified with various SAMs, despite the minor differences in chemical composition identified using standard characterization tools. These different cell morphologies can most probably be explained when investigating the effect of a given SAM layer on the adsorption of proteins present in the culture medium. More likely, it is the ratio between the specific PA22-2 adsorption and nonspecific medium protein adsorption that controls the cellular morphology. Large amounts of adsorbed medium proteins could screen the PA22-2 sites required for cellular attachment. PMID- 19281271 TI - Ubiquitin in NF-kappaB signaling. PMID- 19281273 TI - Optimization of the film-forming and storage conditions of chitosan as an antimicrobial agent. AB - The aim of this work was to assess the antimicrobial capacity of chitosan-based films obtained by a dissolution and solvent evaporation (solvent casting) method at various temperatures (i.e., 37, 80, and 120 degrees C) on the growth of Staphylococcus aureus and Salmonella spp. bacteria. The effect of temperature (4, 23, 37 degrees C) and relative humidity (RH; 0, 75%) during storage on the biocide performance was also investigated. Color parameters and ATR-FTIR spectra were analyzed for each sample to investigate the relationship between structural and/or chemical alterations in the films during storage and biocide performance. The results indicated that films formed at 37 and 80 degrees C presented a significant inhibitory effect for both types of bacteria; however, when cast at 120 degrees C, the films ceased to exhibit antimicrobial properties. Curiously, chitosonium acetate films were seen to diminish to a large extent their biocide properties when stored at 23 degrees C and 75% RH for 2 months or alternatively when stored and 37 degrees C and 0% RH over the same period of time. In good agreement with this behavior the FTIR results indicated that under the previous conditions a significant fraction of the biocide carboxylate chemistry remained in the polymer after contact with the bacterial solution due to a strong reduction in cast film solubility. Because biopolymer active species migration from the film to the culture media is needed for the biomaterial to exhibit measurable antimicrobial effect, proper control of temperature and humidity during film formation and storage is necessary to design the optimum performance of chitosan as a biocide. PMID- 19281274 TI - Surface characteristics of nanocrystalline apatites: effect of mg surface enrichment on morphology, surface hydration species, and cationic environments. AB - The incorporation of foreign ions, such as Mg2+, exhibiting a biological activity for bone regeneration is presently considered as a promising route for increasing the bioactivity of bone-engineering scaffolds. In this work, the morphology, structure, and surface hydration of biomimetic nanocrystalline apatites were investigated before and after surface exchange with such Mg2+ ions, by combining chemical alterations (ion exchange, H2O-D2O exchanges) and physical examinations (Fourier transform infrared spectroscopy (FTIR) and high-resolution transmission electron microscopy (HRTEM)). HRTEM data suggested that the Mg2+/Ca2+ exchange process did not affect the morphology and surface topology of the apatite nanocrystals significantly, while a new phase, likely a hydrated calcium and/or magnesium phosphate, was formed in small amount for high Mg concentrations. Near infrared (NIR) and medium-infrared (MIR) spectroscopies indicated that the samples enriched with Mg2+ were found to retain more water at their surface than the Mg-free sample, both at the level of H2O coordinated to cations and adsorbed in the form of multilayers. Additionally, the H-bonding network in defective subsurface layers was also noticeably modified, indicating that the Mg2+/Ca2+ exchange involved was not limited to the surface. This work is intended to widen the present knowledge on Mg-enriched calcium phosphate-based bioactive materials intended for bone repair applications. PMID- 19281275 TI - Characterization of major radical scavenger species in bovine milk through size exclusion chromatography and functional assays. AB - Radical scavenging activities of bovine milk components were quantified following size exclusion chromatography (SEC) with postcolumn characterization of fractions using the scavenging of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals (ABTS*(+)) in the Trolox equivalent antioxidant capacity (TEAC) assay and peroxyl radicals formed from cleavage of 2,2'-azobis(2-amidinopropane) (AAPH) in the oxygen radical absorbance capacity (ORAC) fluorometric assay. Caseins were quantitatively the major radical scavenger species in both assays, whereas beta lactoglobulin (beta-lg) and alpha-lactalbumin (alpha-la) were much less active and only in the peroxyl radical assay. The radical scavenging activity of the caseins could be quantitatively accounted for by their constituent amino acids, as there were no effects of denaturing agents or complete digestion with proteases. In contrast, the activities of the whey proteins were dependent on denaturation or partial hydrolysis and dominated by the free thiol in beta-lg. A component in milk serum with a molecular mass of approximately 100 kDa contributed significantly to both ABTS*(+) and peroxyl radical scavenging but was absent in whey. This radical scavenger was identified as beta-casein. The only significant low molecular weight radical scavenger species were identified as ascorbate and urate in both assays. PMID- 19281276 TI - Vitamin D2 formation and bioavailability from Agaricus bisporus button mushrooms treated with ultraviolet irradiation. AB - Agaricus bisporus mushrooms contain an abundance of ergosterol, which on exposure to UV irradiation is converted to vitamin D2. The present study evaluated the effects UV-C irradiation on vitamin D2 formation and its bioavailability in rats. Fresh button mushrooms were exposed to UV-C irradiation at mean intensities of 0.403, 0.316, and 0.256 mW/cm(2) from respective distances of 30, 40, and 50 cm for periods ranging from 2.5 to 60 min. Vitamin D2 and ergosterol were measured by HPLC-MS/MS. The stability and retention of vitamin D2 were assessed including the extent of discoloration during storage at 4 degrees C or at room temperature. Exposure to UV-C irradiation at 0.403 mW/cm(2) intensity from 30 cm distance resulted in a time-dependent increase in vitamin D2 concentrations that was significantly higher than those produced at intensities of 0.316 and 0.256 mW/cm(2) from distances of 40 and 50 cm, respectively. Furthermore, the concentrations of vitamin D2 produced after exposure to UV-C irradiation doses of 0.125 and 0.25 J/cm(2) for, 2.5, 5, and 10 min were 6.6, 15.6, and 23.1 microg/g solids, equivalent to 40.6, 95.4, and 141 microg/serving, respectively. The data showed a high rate of conversion from ergosterol to vitamin D2 at short treatment time, which is required by the mushroom industry. The stability of vitamin D2 remained unchanged during storage at 4 degrees C and at room temperature over 8 days (P = 0.36), indicating no degradation of vitamin D2. By visual assessment or using a chromometer, no significant discoloration of irradiated mushrooms, as measured by the degree of "whiteness", was observed when stored at 4 degrees C compared to that observed with mushrooms stored at room temperature over an 8 day period (P < 0.007). Vitamin D2 was well absorbed and metabolized as evidenced by the serum response of 25-hydroxyvitamin D in rats fed the irradiated mushrooms. Taken together, the data suggest that commercial production of button mushrooms enriched with vitamin D2 for improving consumer health may be practical. PMID- 19281277 TI - Feather mites of the genus Proterothrix Gaud (Astigmata: Proctophyllodidae) from parrotbills (Passeriformes: Paradoxornithidae) in China. AB - Three new species of the feather mite genus Proterothrix (Proctophyllodidae: Pterodectinae) are reported from parrotbills (Passeriformes: Paradoxornithidae) in China. We describe Proterothrix paradoxornis n. sp. from Paradoxornis webbianus (type host) and Par. alphonsianus, P. longicaula n. sp. from Par. gularis, and P. sarahbushae n. sp. from Par. verreauxi (type host) and Par. alphonsianus. These are the first records of mites of the Pterodectinae from the Paradoxornithidae. Within Proterothrix, the new species constitutes a distinct complex, characterized by the presence of a long, whip-like aedeagus and spindle shaped setae e on tarsi I in males. PMID- 19281278 TI - A laparoscopic endovascular aortobifemoral conduit that can be retained as a long term bypass: a solution for patients with inadequate iliac access. AB - PURPOSE: To present a laparoscopic technique for placing a partially stented aortobifemoral (ABF) conduit that can be used for more proximal endovascular manipulations and then be retained as a permanent bypass of occlusive iliac disease. TECHNIQUE: Ethical approval was obtained to use a fresh frozen cadaver. The left common iliac artery, distal aorta, and proximal right common iliac artery were dissected laparoscopically. A curved hollow needle was inserted into the distal aorta, and wire access was obtained. A partially stented bifurcated Dacron bypass graft was deployed under fluoroscopic guidance into the distal aorta. The limbs of the bypass were then used as conduits for endovascular access before being tunneled behind the ureters and anastomosed to the femoral arteries in the usual way, retaining the stented graft as an ABF bypass. CONCLUSION: This novel technique combines laparoscopic access with endovascular manipulation to place an ABF conduit, which can be retained as a permanent bypass without the need for an abdominal incision. This technique could provide a minimally invasive solution for pelvic occlusive disease that hinders endovascular repairs, as well as a minimally invasive means of securing endoluminal access in patients with iliac arteries of inadequate caliber. PMID- 19281279 TI - Aortic remodeling in type B aortic dissection: effects of endovascular stent graft repair and medical treatment on true and false lumen volumes. AB - PURPOSE: To analyze aortic remodeling processes in terms of true (TL) and false lumen (FL) volumes in patients with type B aortic dissection undergoing thoracic endovascular aortic repair (TEVAR) versus patients treated medically. METHODS: Serial contrast-enhanced computed tomography (CT) scans of 27 type B dissection patients (24 men; mean age 60+/-13 years) who underwent TEVAR (n = 17) or medical therapy only (n = 10) were analyzed. TL and FL volumes over the entire descending aorta at baseline and at follow-up were quantified by 3-dimensional reconstruction. RESULTS: Follow-up in the 27 patients was a mean 14+/-6 months. TEVAR resulted in a continued gain in total TL volume (132+/-56 mL at baseline, 164+/-51 mL early after TEVAR, and 220+/-68 mL at follow-up, p<0.001), whereas TL volume was almost unchanged in the medical therapy group (113+/-34 to 120+/-41 mL, p = 0.195). Total FL volume decreased significantly in TEVAR patients during follow-up (257+/-147 mL at baseline to 178+/-140 mL, p<0.001), whereas there was no significant change in FL volume in the medical therapy patients. The increase of TL and the decrease of FL volume in the TEVAR group were mainly observed in the descending thoracic aorta. CONCLUSION: TEVAR for type B aortic dissection results in a significant increase in TL and a decrease in FL volumes, not only acutely but also over time due to continued remodeling processes primarily in the thoracic aorta, with little impact on abdominal aortic volumes. Our data provide insight into the mechanism of a potential therapeutic benefit of TEVAR over medical therapy in type B dissection, which remains to be confirmed in a randomized clinical trial. PMID- 19281280 TI - Diameter or volume? The measure of success after endovascular repair of thoracic aortic dissections. PMID- 19281281 TI - Effect of stent-graft compliance on endotension after EVAR. AB - PURPOSE: To investigate whether, and to which extent, the type of graft has influence on the pressure in the aneurysm sac. METHODS: The study consisted of a dynamic and a static experiment. In the dynamic experiment, a latex aneurysm was inserted into an in-vitro circulation model, with variable (80-180 mmHg) systemic systolic pressure and a pulse pressure of 40 mmHg. Seven types of devices were used to exclude the aneurysm: a latex reference graft, 3 stentless grafts made of different fabrics, and 3 different types of commercially produced stented grafts used for endovascular aneurysm repair (EVAR). The systolic and diastolic intra aneurysm pressures (IAP) were measured, along with the pulse pressure. The mean IAP and pulse pressures were compared for each category of graft (stented/stentless) and for each graft. In the static experiments, the compliance of each graft and the latex aneurysm were determined by measuring the change in volume necessary to obtain a change in pressure. Furthermore, the IAP with each graft was measured in a nonpulsatile situation under systolic pressures varying between 60 and 140 mmHg. RESULTS: The experiments showed that, with increasing systemic pressures, there was a small pressure (<5 mmHg) increase in the aneurysm, but there was no significant difference among grafts in the dynamic or the static experiments (p = 0.12 and p = 0.17, respectively). With the 3 stented EVAR grafts, there was less pressure transmission than with the 3 stentless grafts, but this difference was not significant (p>0.05). CONCLUSION: This in vitro study demonstrated that there is a clinically irrelevant pressure transmission through the graft wall and no significant difference in the pressure transmission between stented and stentless grafts. PMID- 19281282 TI - Ultrasonographic surveillance with selective CTA after endovascular repair of abdominal aortic aneurysm. AB - PURPOSE: To evaluate the agreement between color-coded duplex ultrasound (US) and computed tomographic angiography (CTA) in monitoring aneurysm diameter and detecting endoleaks after endovascular aneurysm repair (EVAR). METHODS: From November 1998 to January 2007, 196 patients (191 men; mean age 72.4 years, range 52-88) underwent EVAR and were followed by CTA and US over a mean 3.9+/-2.4 years (range 0-8.9, median 3.4). Annual paired CTA and US examinations were reviewed to assess agreement in measuring maximum aneurysm transverse diameter and to evaluate diagnostic accuracy of US in detecting endoleak. RESULTS: The 5-year cumulative endoleak incidence was 43.8% (72 patients). At first diagnosis, US detected 55/72 (76.4%) endoleaks; of the remaining 17, only 3 (4.3%) were clinically significant in terms of aneurysm enlargement. Pairing 709 annual CTA and US examinations from 184 patients showed a high agreement (k = 0.96) between examinations in measuring maximum transverse diameter, with a mean difference between US and CTA of -2.5 mm. CONCLUSION: After the first year of follow-up, EVAR surveillance costs can be reduced by performing annual US examinations only. Keeping in mind that US underestimates diameter measurements, CTA can be reserved for patients with increasing or persistently stable aneurysm diameters. PMID- 19281283 TI - Safety and performance of targeted renal therapy: the Be-RITe! Registry. AB - PURPOSE: To evaluate the safety and patterns of use of targeted renal therapy (TRT) with the Benephit system. TRT, the delivery of therapeutic agents directly to the kidneys by renal arterial infusion, has the advantage of providing a higher local effective dose with potentially greater renal effects, while limiting systemic adverse effects due to renal first-pass elimination. METHODS: The Benephit System Renal Infusion Therapy (Be-RITe!) Multicenter Registry was a post-market registry following patients treated using the Benephit systems for TRT. The registry enrolled 501 patients (332 men; mean age 72.2+/-9.5 years) at high risk for contrast-induced nephropathy (CIN) during coronary or peripheral angiography/intervention or cardiovascular surgery. The Mehran score was used to compare the actual to predicted incidence of CIN within 48 hours post procedure. RESULTS: Bilateral renal artery cannulation was successful in 94.2%, with a mean cannulation time of 2.0 minutes. Either fenoldopam mesylate, sodium bicarbonate, alprostadil, or B-type natriuretic peptide (BNP) was infused for 184+/-212 minutes. Mean creatinine levels did not change significantly (baseline, 24, and 48 hours post procedure: 1.95, 1.99, and 1.98 mg/dL, respectively; p = NS). In 285 patients who received TRT with fenoldopam and were followed for at least 48 hours, the incidence of CIN was 71% lower than predicted (8.1% actual CIN versus 28.0% predicted; p<0.0001). Only 4 (1.4%) patients required dialysis (versus the 2.6% predicted rate, p = NS). CONCLUSION: The Benephit system and TRT during coronary and endovascular procedures in patients at high risk for renal failure is simple to use and safe. With the infusion of intrarenal fenoldopam, the incidence of CIN was significantly lower than predicted by risk score calculations. PMID- 19281284 TI - Targeted renal therapy: a potential weapon in the battle against CIN. PMID- 19281285 TI - Intraluminal recanalization of long infrainguinal chronic total occlusions using the Crosser system. AB - PURPOSE: To assess the safety and efficacy of a device for vibrational angioplasty in the percutaneous intraluminal recanalization of long infrainguinal chronic total occlusions (CTO). TECHNIQUE: The Crosser CTO Recanalization System is a mechanical recanalization device that uses high-frequency vibrational energy to disrupt and channel through fibrocalcific plaque without harming the vessel wall, thus assisting in the recanalization of an occluded artery. In 12 diabetic patients (7 men; median age 71 years, range 58-80) with critical limb ischemia owing to long (median length 26 cm, range 21-32) infrainguinal CTOs resistant to conventional guidewire techniques, the Crosser CTO Recanalization System was successful in intraluminally crossing the occlusion in 9 (75%) patients in <5 minutes (mean 4:03 minutes). The safety endpoint (distal lumen guidewire position with no vessel perforation or dissection) was achieved in all successful cases. CONCLUSION: In our preliminary experience, the Crosser CTO Recanalization Catheter decreased crossing time, was safe, and achieved a high rate of intraluminal recanalization of long infrainguinal CTOs. PMID- 19281286 TI - Endovascular management of chronic infrarenal aortic occlusion. AB - PURPOSE: To review our experience with the endovascular treatment of chronic infrarenal aortic occlusion with regard to technical success and midterm patency, as well as perioperative mortality and morbidity. METHODS: A retrospective review was performed of patients who presented from January 1, 2000, to December 31, 2005, with a diagnosis of chronic infrarenal aortic occlusion (TASC D) treated with endovascular techniques. In this time period, 31 patients (22 women; mean age 63 years) underwent attempted recanalization of the occluded aorta and iliac arteries. Claudication was the most common presenting symptom (14, 45%). Patients were treated solely with angioplasty and stenting or thrombolysis followed by angioplasty/stenting based on surgeon preference. RESULTS: Technical success was 93%. The 2 failures were individual cases of wire-induced iliac artery perforation and failed access; both patients were treated with bypass grafting. Nine (29%) patients had thrombolysis prior to angioplasty. There were no perioperative deaths. Postoperative ankle-brachial indexes increased significantly from preoperative values (p<0.0001). There were 3 technical complications: 1 (3%) iatrogenic iliac artery injury and 2 (6%) perioperative limb thromboses requiring intervention. Other complications included 6 (19%) access site events and 5 (16%) episodes of acute renal dysfunction, 2 requiring permanent dialysis. Over a mean follow-up of 12 months, there was no limb loss. At 1 and 3 years, the primary/secondary patency rates were 85%/100% and 66%/90%, respectively. CONCLUSION: Endovascular therapy for chronic infrarenal aortic occlusion has a high technical success rate, with good midterm primary and secondary patency rates. However, renal dysfunction can occur; the etiology is likely multifactorial from contrast volumes, embolization, and/or renal arterial disease. PMID- 19281287 TI - Incidence, anatomical location, and clinical significance of compressions and fractures in infrapopliteal balloon-expandable metal stents. AB - PURPOSE: To investigate the incidence, anatomical location, and clinical impact of fractures and/or compression of infrapopliteal balloon-expandable metal stents implanted for critical limb ischemia (CLI) treatment. METHODS: This prospective study included 63 CLI patients (45 men; mean age 71.3+/-9.5 years) who had been treated with infrapopliteal angioplasty and stent placement for 191 lesions in 84 limbs. In all, 369 stents (296 stainless steel and 73 cobalt-chromium alloy) were implanted; 239 were placed overlapping in tandem lesions. Mean length of the overall stented segment was 4.4+/-6.3 cm (range 1.6-14.0). Stents were located in the tibioperoneal (n = 34), anterior tibial (n = 195), posterior tibial (n = 63), and peroneal (n = 77) arteries. Follow-up consisted of digital subtraction angiography and infrapopliteal radiography imaging at 2 different angles. Evaluation of stents for the presence of fracture and/or compression was done after digital processing at the highest possible magnification. Stent fractures were defined according to published standards, whereas compression was classified as severe shape alteration and/or collapse of the stent mesh. Angiographic restenosis was based on a 50% threshold. RESULTS: Mean follow-up was 15+/-11 months (range 6-60). Image analysis detected 1 (0.3%) severe stent fracture (complete separation and misalignment of stent struts) and 11 (3.0%) stent compressions. Infrapopliteal stent fracture and compressions were associated with increased artery restenosis [100% (12/12) versus 47.3% (169/357), p<0.001] and an increased rate of clinical deterioration and clinically-driven reinterventions [41.7% (5/12 limbs) versus 19.4% (14/72 limbs), p = 0.04]. The single fracture and most of the compressions were located in the distal third of the anterior tibial artery. CONCLUSION: Stent fractures and compressions of infrapopliteal balloon-expandable metal stents are infrequent. However, they may be related to increased restenosis. PMID- 19281288 TI - Intentional overstenting of the celiac trunk during thoracic endovascular aortic repair: preoperative role of multislice CT angiography. AB - PURPOSE: To report initial experience with intentional coverage of the celiac trunk to expand the distal landing zone in thoracic endovascular aortic repair (TEVAR) and to analyze preprocedural visualization of collateral blood flow by noninvasive computed tomographic angiography (CTA). METHODS: Between January 1997 and April 2008, 202 patients with thoracic aortic pathologies were treated by TEVAR. In 5 high-risk patients (3 women; mean age 73 years, range 62-88), intentional overstenting of the celiac trunk was performed when treating 2 ruptured aneurysms, 1 penetrating aortic ulcer, 1 type B dissection, and 1 distal type I endoleak. Multislice CTA (MSCTA) with multiplanar reconstruction was used to visualize the visceral collaterals; no angiography was performed. RESULTS: MSCTA was able to visualize the patent pancreaticoduodenal artery and other collaterals and to verify sufficient collateral blood flow. All celiac arteries were patent preoperatively; only 1 severe stenosis was demonstrated. After successful TEVAR in all cases, 1 of the 5 patients developed visceral malperfusion and died of multiorgan failure 1 day after TEVAR of a ruptured thoracic aneurysm. A second patient died 6 weeks postoperatively due to cardiopulmonary failure (non-procedure-related). There were no late complications or reinterventions. CONCLUSION: Overstenting of the celiac trunk is feasible, with acceptable risk in emergency cases and high-risk patients if MSCTA documents collateral blood flow. Noninvasive MSCTA is sufficient and may obviate pre- and intraoperative selective angiography with or without a balloon occlusion test. PMID- 19281289 TI - Imaging to estimate the safety of intentional celiac trunk coverage in TEVAR: multislice CTA cannot replace angiography at present. PMID- 19281290 TI - Treating the thoracic aorta in Marfan syndrome: surgery or TEVAR? AB - Marfan syndrome (MFS) is an inherited disorder of connective tissue that is historically associated with high mortality due to disorders of the cardiovascular system. Over the past 40 years, surgery to the aortic root and thoracoabdominal aorta has resulted in a significantly prolonged life expectancy. More recently, techniques for endovascular repair of the aortic complications of MFS have been described. In this article we review the relative merits of open and endovascular approaches to thoracic aortic complications in MFS, comparing outcomes from contemporary literature. A reasonably large body of evidence has developed concerning the surgical treatment of the aortic root, ascending thoracic aorta, and arch in MFS. It is clear from large series that pathology, such as acute type A dissections and aneurysmal dilatation, can be successfully repaired with low mortality. Recently published series, although reporting smaller numbers, have demonstrated that the same is true for open surgical replacement of descending thoracic and thoracoabdominal aortic aneurysms. The evidence for thoracic endovascular aortic repair in MFS is much more limited. Small series and registry or case reports describe a heterogeneous group of patients with only short-term follow-up. Outcomes of endovascular repair are mixed, with questionable longevity. Reoperation is, however, common in MFS, and minimally invasive techniques may provide a bridging role or alternative solution when revisiting the hostile surgical field. PMID- 19281291 TI - Intermediate to long-term outcomes of endoluminal stent-graft repair in patients with chronic type B aortic dissection. AB - PURPOSE: To assess mid- to long-term clinical results of stent-graft repair in patients with type B aortic dissection. METHODS: Retrospective analysis was done on 72 patients (47 men; mean age 55+/-12 years) who were diagnosed with chronic type B aortic dissection and underwent stent-graft repair from June 1994 to December 2007. Two types of stent-grafts were employed. Prior to 2000, a custom designed stent-graft composed of self-expanding Z-type stainless steel stents covered with woven polytetrafluoroethylene material was used. Thereafter, a separate-type (modular) manufactured stent-graft was used, consisting of proximal and distal nitinol stents with an unsupported Dacron tube graft in between; the unsupported graft was stented after deployment. RESULTS: Procedural success (completion of the stent-graft deployment at the target area without device failure) was obtained in 97% (70/72). The 2 failures were due to stent migration and tortuous anatomy, respectively. There were persistent type I endoleaks in 6 cases for a clinical success (entry tear exclusion) of 88% (64/72). There was no immediate postprocedural mortality or paraplegia. There was unintended partial left subclavian artery occlusion in 1 case, as well as postprocedural transient renal failure in 1 patient and 2 access-site pseudoaneurysms. Median follow-up was 43 months (range 5-97, mean 64.4+/-38.8) in 61 patients (3 patients lost to follow-up). Five patients died, 1 of probable aortic rupture in the setting of residual type I endoleak. Overall clinical success (no death, conversion, or endoleak) was achieved in 84% (47/56); of these, 35 (74%) showed complete resolution of the thoracic false lumen, while the other 12 (26%) had a decrease in false lumen diameter. CONCLUSIONS: Intermediate to long-term results of stent graft implantation in patients with chronic type B aortic dissection seem acceptable and may justify the use of this minimally invasive approach as first line therapy in these patients. PMID- 19281292 TI - Endovascular abdominal aortic aneurysm repair in a patient with severe hemophilia B. AB - PURPOSE: To report a case of endovascular aneurysm repair (EVAR) in a patient with severe hemophilia B. CASE REPORT: A 63-year-old man with severe hemophilia B was referred for the treatment of a symptomatic infrarenal aortic aneurysm that had enlarged from 3.1 to 4.2 cm in diameter in the last 6 months. The patient was successfully treated with EVAR, associated with perioperative administration of titrated doses of high-purity factor IX concentrate. There were no perioperative complications, but a small type II endoleak was found on the initial postoperative computed tomographic scan; on the 6 month scan, the endoleak had not changed, and it is under intensive surveillance. CONCLUSION: This case demonstrates that endovascular abdominal aortic repair is safe and feasible in severe hemophilic patients managed perioperatively with deficient coagulant protein replacement. PMID- 19281293 TI - Dissection of the abdominal aorta. Current evidence and implications for treatment strategies: a review and meta-analysis of 92 patients. AB - PURPOSE: To report meta-analyses of published clinical experiences with abdominal aortic dissection (AAD), a rare event that is accountable for up to 4% of all aortic dissections. METHODS: All English-language articles regarding abdominal aortic dissection were identified using MEDLINE, Cochrane Library Central, and EMBASE databases. All identified articles were critically appraised for relevance and validity before data extraction and the meta-analyses were performed. Original data were identified on 92 AAD patients (62 men; mean age 59+/-16 years). RESULTS: Of all AADs, 73 (79%) were spontaneous, 13 (14%) traumatic, and 6 (7%) iatrogenic; the majority (68, 74%) were acute presentations. The mean dissection length was 59+/-16 mm. A pre-existing abdominal aortic aneurysm was present in 39 (42%). Hypertension was more frequently present in patients with spontaneous dissections compared to other dissection types (p = 0.001). Concurrent aortic aneurysms were more often associated with spontaneous dissections (p = 0.002). Aortic rupture occurred in 9 (10%) AADs. Open surgical repair was performed in 46 (50%), endovascular repair in 19 (21%), and conservative medical treatment in 27 (29%). In-hospital mortality was 4% overall [1 (2%) in the open repair group, 0 endovascular, and 2 (8%) conservative treatment]. Major complications occurred in 9% [6 (13%) in the open repair group, 1 (5%) in the endovascular group, and 1 (4%) in the conservative treatment cohort]. CONCLUSION: AAD is a rare event that appears to be associated with hypertension and pre-existing aneurysmal degeneration of the abdominal aorta. AAD patients are at considerable risk of in-hospital mortality and complications. Endovascular therapy appears to be associated with a relatively low risk of mortality or major complications compared to open repair and conservative treatment. PMID- 19281294 TI - Commentary: a better understanding of the diagnosis and treatment of isolated abdominal aortic dissections. PMID- 19281295 TI - Reptile infection with Anaplasma phagocytophilum, the causative agent of granulocytic anaplasmosis. AB - Granulocytic anaplasmosis (GA) is a potentially fatal tick-borne rickettsial disease that occurs sporadically in the far western United States. We evaluated the prevalence of Anaplasma phagocytophilum in multiple species of lizards and snakes from enzootic sites in northern California, described the infestation prevalence of its tick vector Ixodes pacificus on reptiles, and conducted an experimental challenge of western fence lizards (Sceloporus occidentalis) and Pacific gopher snakes (Pituophis catenifer) with A. phagocytophilum delivered via needle inoculation or tick bite. Both serologically and polymerase-chain reaction (PCR)-positive lizards (seroprevalence = 10.8%, PCR prevalence = 10.2%) and snakes (seroprevalence = 5.8%, PCR prevalence = 11.7%) were detected among wild caught animals. A DNA sequence of the A. phagocytophilum groESL gene from a PCR positive snake was 100% homologous to that of the human-derived A. phagocytophilum. Experimental attempts to infect naive animals were unsuccessful for snakes (n = 2), but 1 of 12 lizards became infected for 1 wk only by tick bite. Xenodiagnostic I. pacificus larvae that fed on a PCR-positive lizard did not acquire or transmit rickettsiae. Our findings suggest that lizards and snakes are exposed to A. phagocytophilum by infected ticks, but that they do not serve as primary reservoir hosts of this rickettsia. PMID- 19281296 TI - Nematodes collected by gastric lavage from live American alligators, Alligator mississippiensis, in Florida. AB - Stomach nematodes were collected from 151 live American alligators, Alligator mississippiensis, from 3 lakes (Apopka, Griffin, and Woodruff) in north-central Florida using a gastric lavage technique. Four species were identified: Dujardinascaris waltoni, Ortleppascaris antipini, Brevimulticaecum tenuicolle, and larvae of Contracaecum sp. Of these, D. waltoni was the most prevalent species in all 3 lakes and was more prevalent in Lake Apopka than in the other 2 lakes. This is the first record of Contracaecum larvae in the American alligator and the second record of O. antipini. PMID- 19281297 TI - Is Gnathostoma turgidum an annual parasite of opossums? Drastic seasonal changes of infection in Didelphis virginiana in Mexico. AB - Gnathostoma turgidum is a nematode that parasitizes the stomach of opossums, Didelphis virginiana. Despite its wide distribution in the Americas, its natural life cycle is poorly understood. Recently, we found an endemic area for G. turgidum infection in Sinaloa, Mexico (Diaz-Camacho et al., 2009). Based on sporadic surveys for several years, the prevalence was apparently high in summer and extremely low in winter. To confirm that this is really a seasonal variance, we conducted a longitudinal survey on G. turgidum infection in opossums from November 2007 to November 2008. The results showed amazing seasonal changes in the prevalence, with synchronized migration and maturation of worms in opossums. Between February and March, many juvenile worms, with occasional AL3, were found in the liver, but no worms were found in the stomach. Mature adult worms began to appear in the stomach around April and rapidly increased in number toward July, when all worms resided in the stomach. Then, the worms disappeared almost completely by November. These results suggest that G. turgidum is an annual parasite of the opossum, D. virginiana, in Mexico. PMID- 19281299 TI - Cross-talk between mitochondria and NADPH oxidase: effects of mild mitochondrial dysfunction on angiotensin II-mediated increase in Nox isoform expression and activity in vascular smooth muscle cells. AB - Mitochondria and NADPH oxidase activation are concomitantly involved in pathogenesis of many vascular diseases. However, possible cross-talk between those ROS-generating systems is unclear. We induced mild mitochondrial dysfunction due to mitochondrial DNA damage after 24 h incubation of rabbit aortic smooth muscle (VSMC) with 250 ng/mL ethidium bromide (EtBr). VSMC remained viable and had 29% less oxygen consumption, 16% greater baseline hydrogen peroxide, and unchanged glutathione levels. Serum-stimulated proliferation was unaltered at 24 h. Although PCR amplification of several mtDNA sequences was preserved, D-Loop mtDNA region showed distinct amplification of shorter products after EtBr. Such evidence for DNA damage was further enhanced after angiotensin II (AngII) incubation. Remarkably, the normally observed increase in VSMC membrane fraction NADPH oxidase activity after AngII was completely abrogated after EtBr, together with failure to upregulate Nox1 mRNA expression. Conversely, basal Nox4 mRNA expression increased 1.6-fold, while being unresponsive to AngII. Similar loss in AngII redox response occurred after 24 h antimycin-A incubation. Enhanced Nox4 expression was unassociated with endoplasmic reticulum stress markers. Protein disulfide isomerase, an NADPH oxidase regulator, exhibited increased expression and inverted pattern of migration to membrane fraction after EtBr. These results unravel functionally relevant cross-talk between mitochondria and NADPH oxidase, which markedly affects redox responses to AngII. PMID- 19281300 TI - Antitumor therapy based on cellular competition. AB - A major obstacle for the efficacy of cancer gene therapy is the need to transduce a high proportion of tumor cells with genes that directly or indirectly cause their death. During the formation of certain organs, cells compete among themselves to colonize the whole tissue. We reasoned that cell competition could be used to increase the proportion of cells that become transfected in a tumor. For this, a transgene that provides a selective advantage to the transfected cells should be used. If the same gene conferred a suicide mechanism the tumor could be eradicated after a period of selection. Bystander effect of transfected cells over neighboring nonmodified cells may eliminate tumors even with incomplete replacement of tumor cells. To test this strategy a competitive advantage was provided to colon cancer cells, using a gene encoding a fusion protein of dihydrofolate reductase (DHFR) and thymidine kinase (TK). DHFR confers resistance to methotrexate (MTX) and TK confers sensitivity to ganciclovir (GCV). Modified cells were also transduced with green fluorescent protein and parental cells with red fluorescent protein. In vitro and in vivo experiments were performed, using various proportions of modified cells and applying positive selection with MTX followed by negative selection with GCV. In vitro, cell competition was evident. Under MTX treatment, tumor cells transfected with the DHFR-TK fusion gene efficiently replaced the parental cells (from 0.1 to 90% in 35 days). After this positive selection period, negative selection with GCV eliminated the transfected cells. In vivo, positive selection was also achieved and resulted in a statistically significant therapeutic effect. PMID- 19281301 TI - Changing challenges of bacterial enteric infection in the United States. PMID- 19281302 TI - Tri-county comprehensive assessment of risk factors for sporadic reportable bacterial enteric infection in children. AB - BACKGROUND: The aim of this study was to determine risk factors for childhood sporadic reportable enteric infection (REI) caused by bacteria, specifically Campylobacter, Salmonella, Escherichia coli O157, or Shigella (REI-B). METHODS: Matched case-control study. Case patients aged <19 years who were reported to 3 Washington State county health departments and matched control subjects were interviewed from November 2003-November 2005. Matched odds ratios (ORs) were calculated by using conditional logistic regression. Population attributable risk percentages were calculated for exposures associated with infection. RESULTS: Two hundred ninety-six case patients were matched to 580 control subjects. Aquatic recreation was the most important factor associated with all REI-Bs studied (beach water exposure [OR for Salmonella infection, 28.3 {CI, 7.2-112.2}; OR for Shigella infection, 14.5 {CI 1.5-141.0} or any recreational water exposure [OR for Campylobacter infection, 2.7 {CI, 1.5-4.8}; OR for Escherichia coli O157 infection, 7.4 {CI, 2.1-26.1}]). Suboptimal kitchen hygiene after preparation of raw meat or chicken (OR, 7.1 [CI, 2.1-24.1]) and consumption of food from restaurants were additional risks for Campylobacter infection. Infection with Salmonella was associated with the use of private wells as sources of drinking water (OR, 6.5 [CI, 1.4-29.7]), and the use of residential septic systems was a risk for both Salmonella (OR, 3.2 [CI, 1.3-7.8]) and E. coli (OR, 5.7 [CI, 1.2 27.2]) O157 infection. CONCLUSIONS: Overall, non-food exposures were as important as food-related exposures with regard to their contributions to the proportion of cases. Infection prevention efforts should address kitchen hygiene practices and non-food exposures, such as recreational water exposure, in addition to food consumption risks. PMID- 19281304 TI - Ability to replicate in the cytoplasm predicts zoonotic transmission of livestock viruses. AB - Understanding viral factors that promote cross-species transmission is important for evaluating the risk of zoonotic emergence. We constructed a database of viruses of domestic artiodactyls and examined the correlation between traits linked in the literature to cross-species transmission and the ability of viruses to infect humans. Among these traits--genomic material, genome segmentation, and replication without nuclear entry--the last is the strongest predictor of cross species transmission. This finding highlights nuclear entry as a barrier to transmission and suggests that the ability to complete replication in the cytoplasm may prove to be a useful indicator of the threat of cross-species transmission. PMID- 19281303 TI - CD73 is expressed by human regulatory T helper cells and suppresses proinflammatory cytokine production and Helicobacter felis-induced gastritis in mice. AB - BACKGROUND: Regulatory T cells (known as "Treg") express apyrases (CD39) and ecto 5'-nucleotidase (CD73) and contribute to their inhibitory function by generating adenosine. We investigated the expression of CD39 and CD73 on human T helper (Th) cells and the role of CD73 in regulating Helicobacter felis-induced gastritis and colonization. METHODS: Human CD4+ Th cells, gastric T cells, or Treg subsets were stimulated and assayed for the expression of CD39 and CD73 by means of reverse transcriptase polymerase chain reaction and flow cytometry. The effect of CD73 on proliferation and cytokine production was assessed, and the presence of gastritis, proinflammatory cytokine expression, or colonization of H. felis was evaluated in CD73-deficient (CD73-/-) mice or recipient mice given control or CD73-/- Treg. RESULTS: CD4+ T cells expressed CD39 and CD73, particularly in CD25+Foxp3+ Treg from peripheral blood or gastric mucosa. Activation significantly increased CD73 expression on all Th cells. Inhibition of CD73 enhanced production of interferon-gamma. Gastritis in H. felis-infected CD73-/- mice was significantly worse than that in wild-type mice and was accompanied by increased levels of proinflammatory cytokines and reduced bacterial colonization, whereas Treg from CD73-/- mice did not inhibit gastritis. CONCLUSION: CD39 and CD73 expressed by Th cells contribute to local accumulation of adenosine and attenuation of gastritis, which may favor persistent infection. PMID- 19281305 TI - Tumor necrosis factor and lymphotoxin-alpha polymorphisms and severe malaria in African populations. AB - The tumor necrosis factor gene (TNF) and lymphotoxin-alpha gene (LTA) have long attracted attention as candidate genes for susceptibility traits for malaria, and several of their polymorphisms have been found to be associated with severe malaria (SM) phenotypes. In a large study involving >10,000 individuals and encompassing 3 African populations, we found evidence to support the reported associations between the TNF -238 polymorphism and SM in The Gambia. However, no TNF/LTA polymorphisms were found to be associated with SM in cohorts in Kenya and Malawi. It has been suggested that the causal polymorphisms regulating the TNF and LTA responses may be located some distance from the genes. Therefore, more detailed mapping of variants across TNF/LTA genes and their flanking regions in the Gambian and allied populations may need to be undertaken to find any causal polymorphisms. PMID- 19281306 TI - Reduced Fc receptor-mediated clearance as explanation of increased antiplasmodial antibody levels in cerebral malaria. PMID- 19281308 TI - Response to "Case of yellow fever vaccine-associated viscerotropic disease with prolonged viremia, robust adaptive immune responses, and polymorphisms in CCR5 and RANTES genes". PMID- 19281309 TI - Nontyphoidal Salmonella serovars cause different degrees of invasive disease globally. PMID- 19281310 TI - The TRPC class of ion channels: a critical review of their roles in slow, sustained increases in intracellular Ca(2+) concentrations. AB - The realization that there exists a multimembered family of cation channels with structural similarity to Drosophila's Trp channel emerged during the second half of the 1990s. In mammals, depending on the species, the TRP family counts 29 or 30 members which has been subdivided into 6 subfamilies on the basis of sequence similarity. TRP channels are nonselective monovalent cation channels, most of which also allow passage of Ca(2+). Many members of each of these families, but not all, are involved in sensory signal transduction. The C-type (for canonical or classical) subfamily, differs from the other TRP subfamilies in that it fulfills two different types of function: membrane depolarization, resembling sensory transduction TRPs, and mediation of sustained increases in intracellular Ca(2+). The mechanism(s) by which the C-class of TRP channels-the TRPCs-are activated is poorly understood and their role in mediating intracellular Ca(2+) increases is being questioned. Both of these questions-mechanism of activation and participation in Ca(2+) entry-are the topics of this review. PMID- 19281311 TI - Mycobacterial subversion of chemotherapeutic reagents and host defense tactics: challenges in tuberculosis drug development. AB - Recent worldwide emergence of multidrug-resistant and extensively drug-resistant tuberculosis is threatening to destabilize tuberculosis control programs and urging global attention to the development of alternative tuberculosis therapies. Major roadblocks limiting the development and effectiveness of new drugs to combat tuberculosis are the profound innate resistance of Mycobacterium tuberculosis to host defense mechanisms as well as its intrinsic tolerance to chemotherapeutic reagents. The triangle of interactions among the pathogen, the host responses, and the drugs used to cure the disease are critical for the outcome of tuberculosis. We must better understand this three-way interaction in order to develop drugs that are able to kill the bacillus in the most effective way and minimize the emergence of drug resistance. Here we review our recent understanding of the molecular basis underlying intrinsic antibiotic resistance and survival tactics of M. tuberculosis. This knowledge may help to reveal current targets for the development of novel antituberculosis drugs. PMID- 19281313 TI - AMWA position statement on abortion and reproductive rights. PMID- 19281314 TI - Ovarian preservation by GnRH agonists during chemotherapy: a meta-analysis. AB - PURPOSE: Treatment with cyclophosphamide (CYC) confers up to a 40% risk of ovarian failure in women of reproductive age. The use of GnRH agonists (GnRHa) to preserve ovarian function has been investigated in several small studies. We performed a systematic review of studies examining whether a GnRHa administered during chemotherapy is protective of ovarian function and fertility. METHODS: We searched the English-language literature (1966-April 2007) using MEDLINE and meeting abstracts and included studies that reported an association between GnRHa and ovarian preservation in women receiving chemotherapy. Studies without a control group were excluded. Ovarian preservation was defined as the resumption of menstrual cycles and a premenopausal follicle-stimulating hormone (FSH) after chemotherapy. Fertility was determined by a woman's ability to become pregnant. We estimated the summary relative risk (RR) and associated 95% confidence intervals (95% CI) using a random-effects model. RESULTS: Nine studies included 366 women. Three studies included women with autoimmune disease receiving CYC; six included women with hematologic malignancy receiving combination chemotherapy. In total, 178 women were treated with GnRHa during chemotherapy, 93% of whom maintained ovarian function. Of the 188 women not treated with GnRHa, 48% maintained ovarian function. The use of a GnRHa during chemotherapy was associated with a 68% increase in the rate of preserved ovarian function compared with women not receiving a GnRHa (summary RR = 1.68, 95% CI 1.34-2.1). Among the GnRHa-treated women, 22% achieved pregnancy following treatment compared with 14% of women without GnRHa therapy (summary RR = 1.65, CI 1.03-2.6). CONCLUSIONS: Based on the available studies, GnRHa appear to improve ovarian function and the ability to achieve pregnancy following chemotherapy. Several randomized trials are underway to define the role and mechanism of GnRHa in ovarian function preservation. In the meantime, premenopausal women facing chemotherapy should be counseled about ovarian preservation options, including the use of GnRHa therapy. PMID- 19281315 TI - Hospitalized women experiencing an episode of excessive oral anticoagulation had a higher bleeding risk than men. AB - OBJECTIVE: Overanticoagulated medical inpatients may be particularly prone to bleeding complications. Among medical inpatients with excessive oral anticoagulation (AC), we sought to identify patient and treatment factors associated with bleeding. METHODS: We prospectively identified consecutive patients receiving oral AC admitted to the medical ward of a university hospital (February-July 2006) who had at least one international normalized ratio (INR) value >3.0 during the hospital stay. We recorded patient characteristics, AC related factors, and concomitant treatments (e.g., platelet inhibitors) that increase the bleeding risk. The outcome was overall bleeding, defined as the occurrence of major or minor bleeding during the hospital stay. We used logistic regression to explore patient and treatment factors associated with bleeding. RESULTS: Overall, 145 inpatients with excessive oral AC comprised our study sample. Atrial fibrillation (59%) and venous thromboembolism (28%) were the most common indications for AC. Twelve patients (8.3%) experienced a bleeding event. Of these, 8 had major bleeding. Women had a somewhat higher risk of major bleeding than men (12.5% vs 4.1%, p = 0.08). Multivariable analysis demonstrated that female gender was independently associated with bleeding (odds ratio [OR] 4.3, 95% confidence interval [95% C1] 1.1-17.8). Age, history of major bleeding, value of the index INR, and concomitant treatment with platelet inhibitors were not independent predictors of bleeding. CONCLUSIONS: We found that hospitalized women experiencing an episode of excessive oral AC have a 4-fold increased risk of bleeding compared with men. Whether overanticoagulated women require more aggressive measures of AC reversal must be examined in further studies. PMID- 19281316 TI - Reproductive and menstrual factors and thyroid cancer among Japanese women: the Japan Collaborative Cohort Study. AB - AIM: Thyroid cancer is more frequent in women than in men, suggesting the potential role of reproductive and menstrual factors in this cancer. To investigate the association with these factors, we examined 37,986 women involved in the Japan Collaborative Cohort (JACC) Study from 1988 to 1997. METHODS: Reproductive and menstrual factors were assessed with a self-administered questionnaire at baseline. Hazard ratios (HR) and 95% confidence intervals (95% CI) of thyroid cancer incidence were estimated using Cox proportional hazards regression. RESULTS: Eighty-six new cases of thyroid cancer were recorded during 379,281 person-years of follow-up. Overall incidence rate was 22.7 per 100,000 person-years, with a diagnosed peak of 38.2 per 100,000 at 55-59 years old. Multivariate HRs of 0.56 (95% CI 0.25-1.24) and 0.52 (95% CI 0.24-1.16) were observed for women who had experienced pregnancy or a live birth, respectively, but without statistical significance. Further, we saw no associations with other factors, such as age at menarche, age at menopause, age at first birth, or hormone use. CONCLUSIONS: There was no significant association between thyroid cancer and reproductive and menstrual factors in the present study. Additional cohort studies should further examine this possible relationship among Japanese women. PMID- 19281317 TI - Domestic violence against women during pregnancy: the case of Palestinian refugees attending an antenatal clinic in Lebanon. AB - OBJECTIVES: To determine the factors associated with domestic violence against pregnant Palestinian refugee women residing in Lebanon and currently using the United Nation Relief and Work Agency's (UNRWA) primary healthcare services. METHODS: This was a cross-sectional study conducted at a polyclinic of primary healthcare of the UNRWA in South Lebanon during the years 2005-2006. The sample was 351 pregnant women who were 15-42 years of age and not accompanied by their husbands or relatives. All women were invited by the midwife to participate in the study during their visit to the clinic for their first checkup or during a follow-up visit. The Abuse Assessment Screen instrument was used to screen for past and recent history of physical and emotional abuse among the participants. RESULTS: Domestic violence was significantly associated with education, gestational age, fear of husband or someone else in the house, and unintended pregnancy. The odds of abuse for women with an elementary or lower education were 6.86 (95% CI 1.2-38.1) and for women with an intermediate or secondary education 6.84 (95% CI 1.4-33.3) compared with women with a university education. The odds of abuse during pregnancy for women whose husbands did not desire their pregnancy were 3.80 (95% CI 1.5-9.7) compared with other women. CONCLUSIONS: Domestic violence against women in Lebanon was associated with educational level, gestational age, fear of husband or someone else in the house, and unintended pregnancy. PMID- 19281318 TI - Adherence to recent screening mammography among Latinas: findings from the California Women's Health Survey. AB - OBJECTIVES: Latinas are less likely than other racial/ethnic groups to engage in mammography rescreening, but little research has examined why Latinas overdue for screening have not had a mammogram recently. This study investigates the factors that affect adherence to screening mammography guidelines among previously screened Latinas. METHODS: Data are from a population-based, random-digit-dial annual probability survey of California women. Participants included 1298 Latinas aged > or =40 who ever had a mammogram. Multivariate logistic regression was used to examine what factors differentiate adherence to recent screening (having a mammogram < or =2 years ago) from being overdue for screening (having a last mammogram >2 years ago). Perceived barriers to screening were also examined among women overdue for a mammogram. Relationships between self-reported perceived barriers to screening and the identified differentiating factors were assessed using chi-square tests. RESULTS: Adherence to recent mammography was reported by 83.9% of previously screened Latinas. When controlling for significant covariates, factors associated with adherence to recent screening included being aged 50-64, having a college education, preferring to interview in Spanish, having health insurance, and reporting a usual source of care (p < 0.05). Lack of affordability was the most frequently cited perceived barrier to screening among Latinas overdue for screening. Age, education, health insurance, and usual source of care were significantly associated with specific perceived barriers to screening mammography. CONCLUSIONS: Results underscore the impact that affordability of, knowledge about, and convenience in obtaining mammograms have on adherence to recent screening. Strategies to improve mammography adherence among Latinas may need to consider that solely improving access to insurance or a regular source of healthcare, although important, may not be sufficient to improve rates of Latinas' screening mammography adherence. PMID- 19281319 TI - Sexual orientation and weight, body image, and weight control practices among young Australian women. AB - OBJECTIVES: We compare weight, body image, and weight control practices of young adult Australian women according to sexual orientation. METHODS: Cross-sectional analyses of the second survey of 9683 young adult women in the Australian Longitudinal Study on Women's Health (ALSWH); the weight, weight control practices, and body image of exclusively heterosexual, mainly heterosexual, bisexual, and lesbian women were compared. RESULTS: Lesbians were less likely to be dissatisfied with their body image (body weight: beta -0.64, 95% CI -1.10- 0.18; body shape: beta -0.83, 95% CI -1.27- -0.40; overall: beta -0.74, 95% CI 1.14- -0.32), to cut down on fats and sugars (OR 0.53, 95% CI 0.34-0.85), and to engage in healthy weight control practices overall (OR 0.48, 95% CI 0.29-0.81) compared with exclusively heterosexual women. Compared with exclusively heterosexual women, bisexual women were more likely to weight cycle (OR 2.22, 95% CI 1.22-4.03). Compared with exclusively heterosexual women, mainly heterosexual and bisexual women were more likely to engage in unhealthy weight control practices overall (mainly heterosexual: OR 1.76, 95% CI 1.42-2.17; bisexuals: OR 2.89, 95% CI 1.57-5.33), such as smoking (mainly heterosexuals: OR 1.83, 95% CI 1.38-2.44; bisexuals: OR 3.80, 95% CI 1.94-7.44) and cutting meals (mainly heterosexuals: OR 1.58, 95% CI 1.23-2.02; bisexual women: OR 3.45, 95% CI 1.82 6.54). Mainly heterosexual women were more likely to vomit (mainly heterosexuals: OR 2.41, 95% CI 1.73-3.36) and use laxatives (mainly heterosexuals: OR 1.56, 95% CI 1.12-2.19). CONCLUSIONS: Future research should explore why bisexual and mainly heterosexual women are at higher risk of disordered eating behaviours. Understanding why lesbians have a healthier body image would also provide insights into how to improve the body image of other groups. It is critical that public health policy and practice address less healthy weight control practices of sexual minority groups. PMID- 19281320 TI - Perceived need for care among low-income immigrant and U.S.-born black and Latina women with depression. AB - PURPOSE: To examine perceived need for care for mental health problems as a possible contributor to ethnic disparities in receiving care among low-income depressed women. METHODS: The role of ethnicity, somatization, and stigma as they relate to perceived need for care is examined. Participants were 1577 low-income women who met criteria for depression. RESULTS: Compared with U.S.-born depressed white women, most depressed ethnic minority women were less likely to perceive a need for mental health care (black immigrants: OR 0.30, p < 0.001; U.S.-born blacks: OR 0.43, p < 0.001; immigrant Latinas: OR 0.52, p < 0.01). Stigma-related concerns decreased the likelihood of perceiving a need for mental health care (OR 0.80, p < 0.05). Having multiple somatic symptoms (OR 1.57, p < 0.001) increased the likelihood of endorsing perceived need. CONCLUSIONS: Findings suggest that there are ethnic differences in perceived need for mental healthcare that may partially account for the low rates of care for depression among low-income and minority women. The relations among stigma, somatization, and perceived need were strikingly similar across ethnic groups. PMID- 19281321 TI - Randomized trial of circular muscle versus pelvic floor training for stress urinary incontinence in women. AB - BACKGROUND: Conservative management, such as pelvic floor muscle training (PMFT), is commonly recommended as first-line therapy for women with stress urinary incontinence (SUI). METHODS: We randomly assigned 245 women with SUI to 12 weeks of circular muscle exercises (Paula method) or PMFT in order to assess whether these approaches are equivalent. End points after 12 weeks included urinary leak as measured by a 1-hour pad test, subjective assessment of incontinence, and quality of life (QOL). Cure was defined as urinary leakage of <1 g. RESULTS: The mean decrease in urinary leakage was 7.9 g (SD 12.1) among women in the Paula group and 8.9 g (SD 18.2) in the PFMT group (90% confidence interval [CI] of between-group difference was -4.68 g to 3.0 g). This did not meet the prespecified criterion for equivalence. There were 15.2% (p = 0.04) more cures in those randomized to the Paula method. Improvement in subjective urinary complaints and QOL was observed in both groups. The study was limited by a dropout rate of 26.6%. CONCLUSIONS: Both methods are efficacious in women with SUI. The results suggest superiority of the Paula method in terms of cure rate. PMID- 19281322 TI - Current contraceptive awareness and use in Greek dental school students. AB - AIMS: We conducted a pilot study in female dental school students in Northern Greece in order to assess their awareness and practice of contraception. METHODS: The study population consisted of 88 female graduating students of the dental school. A self-administered, anonymous questionnaire was designed to explore students' awareness and use of contraceptive methods. RESULTS: Condoms were the most widely used contraceptive method (they were used by 52.3% of the students); 20.5% of the students were using condoms in alternation or together with coitus interruptus, and 6.8% were using only coitus interruptus. The oral contraceptive pill (OC) was used as the only contraceptive method by 4.5% of the students and in combination with condoms by 9.1% of them. The majority (53.4%) considered condoms as the most effective contraceptive method; 9.1% of the students answered "sterilization" and the same percentage stated "intrauterine contraceptive device (IUD)." Overall, 20.5% of the students believed that the OC increases the risk of cancer, and 36.4% did not know if there is an association between OC use and risk of cancer. Among students not using OC, the respective percentages were 23.7% and 42.1%. among the students, 59.1% had asked their gynecologist about contraception. The contraceptive method used and the perception regarding the most effective contraceptive method did not differ significantly between those who had consulted their gynecologist and those who had not. CONCLUSIONS: Dental school students in Greece appear to have inaccurate knowledge on important contraceptive issues, and this is reflected in their contraceptive practices. There is a pressing need to provide scientifically based sexual education if we are to avoid unwanted pregnancies. PMID- 19281323 TI - The HPV vaccine: a content analysis of online news stories. AB - PURPOSE: Approximately 73 million adults in the United States report using the Internet as a source for health information. This study examines the quality, content, and scope of human papillomavirus (HPV) vaccine Internet news coverage starting on the day of its licensure. Information about the HPV vaccine in the media may influence personal attitudes and vaccine uptake. METHODS: Using four search engines and six search terms, a sample of 250 Internet articles on the HPV vaccine were identified between June 8, 2006, and September 26, 2006. The coding instrument captured how the headline was depicted and how the vaccine was labeled in addition to information about HPV, cervical cancer, the HPV vaccine, and current social issues and concerns about the vaccine. RESULTS: Analysis revealed balanced Internet news coverage; 52.4% of Internet news stories were coded as neutral toward the vaccine. Eighty-eight percent of articles labeled the vaccine as a cervical cancer vaccine; 73.5% explained the link between HPV and cervical cancer, although without providing background information on HPV or cervical cancer. Vaccine affordability was the most cited social concern (49.2%). Information about vaccine safety and side effects, duration of vaccine protection, and availability of the catchup vaccine for females aged 13-26 was repeatedly missing. CONCLUSIONS: The HPV vaccine is being marketed as a vaccine to prevent cervical cancer. Comprehensive information on the vaccine, HPV, and cervical cancer continues to be missing from media coverage. Public health educators should monitor online media in an effort to respond to inaccurate information. Barriers to vaccine cost and funding mechanisms need to be addressed more effectively by states. Knowledge of particular media messages could provide a starting point for tackling opposition and uptake issues for future sexually transmitted infection (STI) vaccines. PMID- 19281324 TI - A lifestyle approach to reducing cardiovascular risk factors in underserved women: design and methods of the Illinois WISEWOMAN Program. AB - BACKGROUND: Few lifestyle intervention programs address the needs of financially disadvantaged, low literacy populations. The overall goal of the Illinois WISEWOMAN Program (IWP) was to design such a program and test its effectiveness in reducing cardiovascular disease (CVD) risk, specifically physical activity and nutrition factors. The purpose of this paper is to describe the IWP study design and methods, development of the evidence-based curriculum appropriate for a low socioeconomic status (SES) population, and baseline characteristics of IWP participants. METHODS: The Cooper Institute, in collaboration with the Illinois Department of Public Health and the University of Illinois at Chicago, adapted evidence-based interventions for financially disadvantaged, low literacy populations. The study used a randomized, two-group, experimental design. In total, 1021 women were recruited from the Illinois Breast and Cervical Cancer Program, which serves uninsured and underinsured women, aged 40-64, at or below 200% of poverty. The women were randomized to either a minimum intervention (MI) or an enhanced intervention (EI) group. Both groups received CVD risk factor screening and educational materials. Additionally, the EI group received a 12 week lifestyle intervention. RESULTS: Baseline comparisons show equivalent groups. IWP participants had a higher prevalence of obesity and smoking than similar national samples. CONCLUSIONS: IWP addressed many of the cultural and implementation barriers in programs that seek to improve the health of financially disadvantaged, low literacy populations. Because of the high burden of disease, the unique study population, and the sound design, we anticipate that our future results will contribute to the translation literature, which has largely ignored significant health disparities. PMID- 19281326 TI - Pluripotent stem cells derived from adult human testes. AB - Recent reports have demonstrated that adult tissue cells can be induced to pluripotency, the iPS cells, mostly with the addition of genes delivered using viruses. Also, several publications both in mouse and in human have demonstrated that spermatogonial stem cells (SSCs) from testes can convert back to embryonic stem (ES)-like cells without the addition of genes. Furthermore, these pluripotent ES-like cells can differentiate into all three germ layers and organ lineages. Thus, SSCs have great potential for cell-based, autologous organ regeneration therapy for various diseases. We obtained testes from organ donors and using 1 g pieces of tissue (biopsy size) we demonstrate that testis germ cells (putative SSCs and/or their progenitors) reprogram to pluripotency when removed from their stem cell niche and when appropriate growth factors and reagents in embryonic stem cell medium are added. In addition, our method of obtaining pluripotent ES-like cells from germ cells is simpler than the described methods and may be more suitable if this procedure is developed for the clinic to obtain pluripotent cells to cure disease. PMID- 19281327 TI - Empirical versus preemptive antifungal therapy for high-risk, febrile, neutropenic patients: a randomized, controlled trial. AB - BACKGROUND: Empirical antifungal therapy is the standard of care for neutropenic patients with hematological malignancies who remain febrile despite broad spectrum antibacterial treatment. Recent diagnostic improvements may ensure the early diagnosis of potentially invasive fungal disease. Reserving antifungals for this stage may achieve similar survival rates and reduce treatment toxicity and costs. METHODS: In this multicenter, open-label, randomized noninferiority trial, we compared an empirical antifungal strategy with a preemptive one. Empirical treatment was defined as antibacterial treatment of patients who have persistent or recurrent fever. Preemptive treatment was defined as treatment of patients who have clinical, imaging, or galactomannan-antigen-assay evidence suggesting fungal disease. First-line antifungal treatment was amphotericin B deoxycholate (1 mg/kg/day) or liposomal amphotericin (3 mg/kg/day), depending on daily renal function. The primary efficacy outcome was the proportion of patients alive at 14 days after recovery from neutropenia. RESULTS: The median duration of neutropenia (neutrophil count, <500 cells/mm3) for the 293 patients enrolled was 18 days (range, 5-69 days). By intention-to-treat analysis, survival was 97.3% with empirical treatment and 95.1% with preemptive treatment. The lower 95% confidence limit for the difference in mortality was -5.9%, which was within the noninferiority margin of -8%. Probable or proven invasive fungal infections were more common among patients who received preemptive treatment than among patients who received empirical treatment (13 of 143 vs. 4 of 150; P < .05), and most infections occurred during induction therapy (12 of 73 patients in the preemptive treatment group vs. 3 of 78 patients in the empirical treatment group were infected during induction therapy; P < .01). Preemptive treatment did not decrease nephrotoxicity but decreased costs of antifungal therapy by 35%. CONCLUSIONS: Preemptive treatment increased the incidence of invasive fungal disease, without increasing mortality, and decreased the costs of antifungal drugs. Empirical treatment may provide better survival rates for patients receiving induction chemotherapy. PMID- 19281328 TI - Multistate outbreak of Salmonella Typhimurium and Saintpaul infections associated with unpasteurized orange juice--United States, 2005. AB - BACKGROUND: Infection due to Salmonella species causes an estimated 1.4 million illnesses and 400 deaths annually in the United States. Orange juice is a known vehicle of salmonellosis, for which regulatory controls have recently been implemented. We investigated a nationwide outbreak of Salmonella infection to determine the magnitude of the outbreak and to identify risk factors for infection. METHODS: We identified cases through national laboratory-based surveillance. In a case-control study, we defined a case as infection with Salmonella serotype Typhimurium that demonstrated the outbreak pulsed-field gel electrophoresis pattern in a person with illness onset from 1 May through 31 July 2005; control subjects were identified through random digit dialing. RESULTS: We identified 152 cases in 23 states. Detailed information was available for 95 cases. The median age of patients was 23 years; 46 (48%) of the 95 patients were female. For 38 patients and 53 age-group matched control subjects in 5 states, illness was associated with consuming orange juice (90% vs. 43%; odds ratio, 22.2; 95% confidence interval, 3.5-927.5). In a conditional logistic regression model, illness was associated with consuming unpasteurized orange juice from company X (53% vs. 0%; odds ratio, 38.0; 95% confidence interval, 6.5-infinity). The US Food and Drug Administration found that company X was noncompliant with the juice Hazard Analysis and Critical Control Point regulation and isolated Salmonella serotype Saintpaul from company X's orange juice. CONCLUSIONS: Unpasteurized orange juice from company X was the vehicle of a widespread outbreak of salmonellosis. Although the route of contamination is unknown, noncompliance with the juice Hazard Analysis and Critical Control Point regulation likely contributed to this outbreak. Pasteurization or other reliable treatment of orange juice could prevent similar outbreaks. PMID- 19281329 TI - Timely intervention for invasive fungal disease: should the road now lead to the laboratory instead of the pharmacy? PMID- 19281330 TI - Immunogenicity, safety, and cross-reactivity of an inactivated, adjuvanted, prototype pandemic influenza (H5N1) vaccine: a phase II, double-blind, randomized trial. AB - BACKGROUND: Avian influenza A virus H5N1 has the potential to cause a pandemic. Adjuvants and whole-virion vaccines are regarded as antigen sparing for pandemic vaccines. METHODS: A double-blind, randomized trial was performed from 28 August to 22 December 2007 in 402 adults; 301 adults were randomly assigned to receive 2 doses of an inactivated, aluminum-adjuvanted, whole-virion H5N1 vaccine containing 5, 10, or 15 microg of hemagglutinin per dose 28 days apart, and 101 of them received 2 doses of 10 microg of vaccine 14 days apart. The vaccine was manufactured from the recombinant A/Vietman/1194/2004 (NIBRG14) strain. Blood samples were collected for hemagglutination inhibition and microneutralization assays. RESULTS: All formulations were well tolerated, with no serious adverse events. Most local and systemic reactions were mild or moderate. Immune responses were induced after 1 dose in all vaccination groups. The highest immune response was seen after 2 doses of 15 microg of vaccine, with 90% and 100% seroconversion rates and 90% and 100% of participants having a titer of > or = 1:40 for hemagglutination inhibition and microneutralization assays, respectively. Both the 10- and 15-microg doses met or exceeded European Union licensure criteria. Generally, higher immune responses were elicited in participants vaccinated 28 days apart than those vaccinated 14 days apart. Cross-reaction assays showed that after 2 doses of 10 microg of vaccine, 98% and 87% of participants had a microneutralization titer of > or = 1:40 against heterologous Indonesia and Anhui strains, respectively. CONCLUSIONS: The inactivated, aluminum-adjuvanted, whole virion H5N1 vaccine not only showed good immunogenicity and safety but also elicited significant cross-reactivity against heterologous H5N1 strains in clade 2. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT00535665. PMID- 19281333 TI - A novel antibody to oxidized phosphatidylethanolamine that is specific for amino groups modified by lipid peroxides. AB - ABSTRACT In this study, a novel antibody to oxidized phosphatidylethanolamine (PtdEtn) was generated and characterized. Oxidized PtdEtn antiserum recognized PtdEtn and not unoxidized. PtdEtn by ELISA. The antibody also immunstained cells previously incubated with lipid peroxides (LOOH). This antibody recognized only high-molecular-weight epitopes in plasma. Because this antibody recognizes a specific amino group modification without the accompanying alterations in amino acids that are usually encountered when proteins are incubated with LOOH, it might be useful for the selective detection of such epitopes in cells, tissues, and lipoproteins. PMID- 19281331 TI - Seasonal influenza in adults and children--diagnosis, treatment, chemoprophylaxis, and institutional outbreak management: clinical practice guidelines of the Infectious Diseases Society of America. AB - Guidelines for the treatment of persons with influenza virus infection were prepared by an Expert Panel of the Infectious Diseases Society of America. The evidence-based guidelines encompass diagnostic issues, treatment and chemoprophylaxis with antiviral medications, and issues related to institutional outbreak management for seasonal (interpandemic) influenza. They are intended for use by physicians in all medical specialties with direct patient care, because influenza virus infection is common in communities during influenza season and may be encountered by practitioners caring for a wide variety of patients. PMID- 19281334 TI - Effect of spirulina on the secretion of cytokines from peripheral blood mononuclear cells. AB - ABSTRACT The purpose of this study was to evaluate the immunomodulatory activity of Spirulina, a bluegreen alga used as a food supplement. The effects of Spirulina on the secretion of three cytokines from unstimulated and stimulated human peripheral blood mononuclear cells (PBMC) were examined. In resting PBMC, Spirulina stimulated secretion of interleukin (IL)-1beta, IL-4, and interferon (IFN)-gamma to nearly 2.0, 3.3, and 13.6 times basal levels, respectively. Spirulina induced levels of IFN-gamma (229 +/- 104 pg/ml) that were comparable to those seen after phytohemagglutinin (PHA) stimulation (476 +/- 121 pg/ml). However, it was much less mitogenic than PHA (13.1 +/- 6.9 pg/ml) with respect to the induction of IL-4 secretion (0.34 +/- 0.1 pg/ml). In PHA-stimulated cells, Spirulina enhanced secretion of IL-1beta, IL-4, and IFN-beta by 2.9, 4.0., and 1.6 times, respectively. Although Spirulina stimulates several cytokines, it is clearly more effective in the generation of a Thl-type response. This in vitro study offers additional data for consideration of the potential therapeutic benefits of Spirulina. PMID- 19281335 TI - Cranberry juice induces nitric oxide-dependent vasodilation in vitro and its infusion transiently reduces blood pressure in anesthetized rats. AB - ABSTRACT Red wine vasodilates rat aortae, an effect attributed to polyphenolic compounds. Cranberry juice (CBJ) is also rich in polyphenols. We determined that CBJ has vasorelaxing properties similar to those of red wine. Rat aortic rings cleaned in Krebs buffer, pH 7.4, bubbled with 95% O(2) and 5% CO(2) were recovered for 30 minutes at 37 degrees C under 2.0 g tension. After phenylephrine (PE, 100 mumol/L) contraction, acetylcholine (3 mumol/L)-induced relaxation of intact vessel was significantly higher than in denuded vessels (59.1 +/- 0.27% versus 10.1 +/- 0.09% of the maximal PE contraction; P <.003). After a second PE contraction, a 1:100 dilution of CBJ was added. Intact rings were vasodilated by CBJ with 56.7 +/- 0.26% relaxation, compared to denuded rings with 8.9 +/- 0.06% relaxation (P <.002). Addition of L-NAME reversed CBJ-induced vasorelaxation in intact vessels with 0.54 +/- 0.34 g compared to 0.04 +/- 0.04 g in denuded vessels (P <.007). Subsequent addition of L-arginine resulted in a return of vasodilation in intact vessels. Additionally, CBJ infusion at a 1:100 dilution of estimated blood volume resulted in a 16% reduction of mean arterial blood pressure in anesthetized rats. This study suggests that, like red wine, CBJ has the capacity to exert in vitro and in vivo vasodilatory effects. PMID- 19281337 TI - Wild lettuce: a bitter herb of biblical proportions. PMID- 19281336 TI - Phloretin glycosides: bioactive compounds in apple fruit, purees, and juices. AB - ABSTRACT Phloretin glycosides are bioactive compounds naturally present in apple fruits. The highest amount of phloretin glycosides was significantly found in apple pureees and commercial juices as a consequence of the processing conditions. PMID- 19281338 TI - GMOs: The Court and Our Health. PMID- 19281339 TI - Whey protein concentrates with and without immunoglobulins: a review. AB - ABSTRACT Whey protein, a high-biological-value protein from milk, is available with and without immunoglobulins. The possibility of providing passive immunization through food is unique to mammals, who receive it through colostrum at birth. Colostrum contains both immunostimulatory and immunosuppressive immunoglobulins. Later, the mammals are able to make their own immunoglobulins, but in time of illness that ability may become impaired. Some of the whey proteins contain bovine immunoglobulins, which are similar to human immunoglobulins. This type of whey may be of clinical importance to patients who are immunocompromised. However, all whey proteins are rich in cysteine and glutamate. These serve as precursors of glutathione, a potent antioxidant, which is needed in greater quantities during stress. In this review, the benefits of consuming whey protein with and without immunoglobulins are examined. Characteristics of these products are discussed, and an account is given of supportive studies using the whey containing immunoglobulins in the clinical setting. Patients with acquired immunodeficiency syndrome and cancer are used as examples. Finally, a guide on use of whey protein is provided. PMID- 19281340 TI - Bioabsorption and in vivo antioxidant properties of grape extract biovin((r)): a human intervention study. AB - ABSTRACT Recent epidemiological studies have indicated that intake of grapes and other food products such as red wine derived from grapes may lower incidence of cancer and coronary heart diseases. Grapes and grape extracts, in addition to the traditional nutrients such as sugars and vitamins, are also rich sources of polyphenolic antioxidants. A study was undertaken with the overall objective of evaluating the antioxidant potential of the grape extract Biovin((R)) (Cyvex Nutrition, Inc., Irvine, CA) both in vitro and in healthy human subjects. A dose response relationship between the grape extract and its antioxidant potential was observed in the in vitro and in liposome system. Fourteen subjects (7 men and 7 women) were orally given Biovin, 375 mg/day for 4 weeks, followed by a 2-week washout period during which consumption of Biovin was discontinued. Fasting blood samples were collected at the beginning and end of the treatment period and at the end of the posttreatment washout period. Serum polyphenols, antioxidants, and biomarkers of oxidation including lipid, protein, and low-density lipoprotein (LDL) oxidation were measured. Results showed that Biovin polyphenols were well tolerated by the subjects and were absorbed readily. On discontinuation of consumption of the Biovin capsules, serum levels of the polyphenols were depleted rapidly. Consumption of Biovin was also associated with increased serum antioxidant potential and lower protein and LDL oxidation. Levels of other antioxidants measured did not differ with the treatment. Results from this study strongly suggest an important beneficial role for Biovin polyphenols in enhancing the antioxidant capacity of the serum and protecting proteins and LDL cholesterol against oxidative damage. These outcomes can significantly reduce the risk of chronic diseases such as cancer and coronary heart diseases. PMID- 19281341 TI - Regulatory issues relating to herbal products-part 1: legislation in the European union, north america, and australia. AB - ABSTRACT The increased public awareness about herbal products has led to a growth of the botanicals market and attracted the interest of the pharmaceutical industry. The laws that regulate the sale of manufactured herbal products differ between regions and countries of the Western world. The European Union favors the classification of the remedies as medicinal products. However, differences in the interpretation of European law by the Member States have meant that herbal products have been assigned to various sale categories across the European market. The European Agency for the Evaluation of Medicinal Products is working to introduce changes to achieve harmonization within the market. Canada and Australia have embarked on similar reforms to improve product regulation without imposing undue restrictions on marketing. In these countries, herbal products have been assigned a status that respects their therapeutic role. Changes introduced in the United States of America in 1994 led to a deregulation of the market with herbal products being classified as dietary supplements and excluded from the stricter regulatory requirements applicable to drugs and foods. However, a United States Presidential Commission has advised that a study is needed regarding an alternative system for botanical products that are not used solely as dietary supplements but that cannot meet the requirements applicable for conventional over-the-counter drugs. PMID- 19281342 TI - Regulatory issues relating to herbal products-part 2: safety and toxicity. AB - ABSTRACT A significant proportion of the general public has the misconception that herbal products are safe because they are of natural origin. Toxicity may arise from inherent properties of the herbal ingredients that result in adverse reactions, which may be serious, or from interactions with conventional drug substances. Toxicity may also be caused by misuse, abuse, and overuse of products and by adulteration of products or misidentification of plants. Regulatory authorities are undertaking to provide a reasonable assurance of efficacy and improved safety of herbal products through monitoring programs and control of herbs that are known to be toxic. Action is taken by the authorities to protect public health when herbal preparations are shown or suspected to cause harm. Case reports of toxicity due to herbal products and actions taken by regulatory authorities are summarized in this article. In the European Union, applicants for marketing authorizations may be required to provide scientific proof of safety and efficacy by reference to bibliographic data, where available. The relevance and credibility of published data needs to be assessed against recommended criteria in order to avoid unnecessary repetition of tests on animals or humans. PMID- 19281343 TI - Regulatory issues relating to herbal products-part 3: quality and its determination. AB - ABSTRACT The concept of quality in the manufacture of herbal products is directed toward ensuring that the required standard of product is attained through processes that involve good manufacturing practice and quality control. This article reviews the need for quality assurance of herbal products, approaches taken by regulatory authorities in Europe to address this issue, and analytical methods applicable to the evaluation of herbal products. The natural ingredients that are used to prepare herbal products give rise to problems that are not encountered with conventional medicinal products. Quality standards are applied to both starting materials and finished products. Pharmacopoeial monographs, even when not forming part of official compendia, lay down criteria for analytical testing, including materials and methods, test limits, and acceptance ranges for results. Analytical tests are employed to establish the identity of the plant material or of the active principles, the content of active principles or characteristic marker compounds by assay, and the purity of the product by exclusion of specific adulterants or contaminants. Advances in instrumentation and test methods provide the analyst with powerful tools to determine the level of quality. Nevertheless, there still exist limitations to testing because of the complex and variable nature of the herbal ingredients. The standards and requirements for quality of herbal products as applied in the European Union are also intended to provide further assurances on the efficacy and safety of products. PMID- 19281345 TI - A routine HIV screening program in a South Carolina community health center in an area of low HIV prevalence. AB - In 2006, the Centers for Disease Control and Prevention published guidelines for routine HIV screening in healthcare settings. Feasibility studies have demonstrated that screening is effective in high-volume, urban settings, but there are no data for smaller, more rural settings. The main objective of this study was to describe a routine HIV screening program at a community health center in South Carolina serving both urban and rural populations. Margaret J. Weston Community Health Center implemented routine HIV screening using rapid tests at its three locations on December 1, 2006. All individuals utilizing this center over the age of 13 years were screened for HIV unless they opted out. Nurses completed a survey about their experiences with the program. chi(2) tests and logistic regression models were used to analyze the data. In the first 8 months, among 985 eligible visits, 574 (58%) resulted in the patient being screened. The most common reason for refusal was "doesn't think s/he is at risk." Acceptance rates differed significantly by location (p = 0.01), from 62% in the urban site to 47% in the rural site. Other significant predictors of accepting HIV testing were race/ethnicity, age, and method of payment. Three hundred twenty four (58%) individuals who were tested reported no history of being previously tested for HIV infection. Participation in the screening program was perceived favorably by nurses. This pilot project in a South Carolina community health center demonstrates that implementation of routine HIV screening is acceptable in small healthcare settings and in smaller cities and rural communities in the South. PMID- 19281346 TI - Chronicity and remission of fatigue in patients with established HIV infection. AB - Fatigue is one of the most common and debilitating complaints of HIV-positive individuals, potentially leading to important functional limitations. We recruited 128 HIV-positive individuals (fatigued and nonfatigued) between March 2005 and May 2006; 66% were male, 66% were African American, 45% had greater than a high school education, 67% were unemployed, and ages ranged from 26-66 (median, 44). Every 3 months for 15 months, participants completed a 56-item self-report fatigue scale developed and validated by the authors. Participants were classified as fatigued or not fatigued at each assessment and received scores for fatigue intensity and impact of fatigue on functioning. We used linear mixed effects models to assess longitudinal variation in fatigue scores and generalized estimating equations for binary outcomes to model predictors of fatigue remission among those fatigued at baseline. At baseline, 88% of the sample was fatigued. Fatigue measures were highly correlated across time points (rho 0.63-0.85 [intensity], 0.63-0.80 [functioning]) and showed no evidence of overall improvement, deterioration, or convergence over time. Predictors of lower fatigue scores included higher income, employment, longer time since HIV diagnosis, and antiretroviral therapy use. Those employed at baseline were likely to show improvements in fatigue while those unemployed were not. Of those fatigued at baseline, 11% experienced remission during follow-up; remission was associated with Caucasian race and employment. In summary, fatigue intensity and related functional limitations were persistent, stable, and unlikely to remit over 15 months of follow-up in this sample of patients with established HIV infection. PMID- 19281347 TI - Chemical and Genetic Evaluation of Somaclonal Variants of Egyptian Garlic (Allium sativum L.). AB - Garlic (Allium sativum L.) is used in the household and as an ingredient in many pharmaceutical products. Tissue culture technique provides an excellent source for induction of both chemical and genetic variation in garlic. A callus was induced on root meristem cultured on Murashige and Skoog (MS) medium in the presence of kinetin, indole acetic acid, and 2,4-dichlorophenoxyacetic acid. Shoots with a small bulb were produced on medium containing MS salts, B vitamins, and naphthalene acetic acid. Regenerated plants were transplanted into soil, and a nondivided bulb was formed in the first somaclonal generation (SCI). Plants were normal in their phenotypes in SC2. After four cycles of field cultivation, the selected somaclones (variants) in the fourth generation showed significant differences in bulb character compared with the original plants. Mitotic division and chromosomal abnormalities were investigated in meristimic root tip cells of regenerated plants for the first and fourth regeneration and for control plants. Somaclonal variant metaphase cells had the same chromosome number (2n = 16) as those of the controls. Allicin was measured quantitatively in the regenerated clones by high-performance liquid chromatography. The results showed that some clones contained as much as 14.50 mg/g allicin, compared with 3.80 mg/g in the control plant. This finding suggests that this technique may be useful to improve the allicin content of Egyptian garlic, which could be utilized as a good source for garlic-containing pharmaceutical preparations. PMID- 19281348 TI - A Comparative Study of the Constituents of Aesculus hippocastanum and Aesculus indica. AB - This work compared the phytochemical composition of two species, of Aesculus hippocastanum (horse chestnut) and Aesculus indica grown under comparable conditions. Thin-layer chromatography zone profiles of the constituents in methanol extracts from leaves, seeds, and husks indicated differences within and between the two species. Similar profiles were observed for seeds and leaves from the two species but not for the husks. Aesculin and its aglycone, aesculetin, were not detected in leaves, husks, or seeds of the two species. The content of aescin found in A. indica seeds (13.4% weight/weight) was higher than in A. hippocastanum (9.5%). Laurie acid, palmitic acid, myristic acid, stearic acid, arachidic acid, and oleic acid were detected, by capillary gas chromatography, in the saponified extracts of the seeds, leaves, and husks of the two species. PMID- 19281349 TI - Pilot study on the effect of hyperimmune egg protein on elevated cholesterol levels and cardiovascular risk factors. AB - Coronary heart disease is the leading cause of death in the United States, accounting for almost half of all deaths. Animal studies have suggested that the daily addition of "hyperimmune egg" to one's diet might reduce the risk of cardiovascular disease. Military personnel with initial total cholesterol levels higher than 180 mg/dl were randomly enrolled in a 26-week double-blind study of a drink containing a hyperimmune egg protein. Subjects were randomly assigned to three groups: control (no drink); placebo (drink without egg); and active (drink combined with hyperimmune egg). Throughout the study this physically fit group maintained a program of strenuous exercise and participated in a dietary education program intended to reduce fat and cholesterol intake. At the end of the trial, total cholesterol levels of the control and placebo groups had increased, whereas the group that consumed the drink with hyperimmune egg showed no significant change in total cholesterol. The ratio of total cholesterol to high-density lipoproteins and the apolipoprotein B level increased in both control and placebo groups but remained essentially unchanged in the group consuming hyperimmune egg. Triglyceride and apolipoprotein A-I values did not change significantly in any of the groups. These findings suggest that hyperimmune egg may beneficially modify the regulation of serum lipoprotein levels and thereby reduce the possibility of cardiovascular diseases. PMID- 19281350 TI - Availability of magnesium from various commercially available oral preparations in a variety of dosage forms: comparative in vitro study. AB - The relative dialysability of magnesium in a number of different inorganic and organic magnesium-containing compounds from ten commercially available products was investigated using an in vitro method. Reference values were provided by tests carried out in parallel using comparable quantities of pure magnesium compounds, as contained in the products. The results demonstrated that the excipients generally had a positive effect on the relative availability of magnesium. Furthermore, the dialysed magnesium levels were lower in capsules and coated tablets than in granulates, chewable tablets, and effervescent tablets. It can be concluded that substances such as citric acid, lactose, and sucrose have a positive effect, whereas gel-based excipients and coatings have a negative effect on the availability of magnesium. PMID- 19281352 TI - Newswatch. PMID- 19281351 TI - The Herbal Sling vs. the Magic Bullet. PMID- 19281354 TI - A crisis in the soy fields. PMID- 19281355 TI - Symposium on phytoestrogen research methods: chemistry, analysis, and biological properties. PMID- 19281356 TI - Biosynthesis and distribution of phytoestrogens and their roles in plant defense, signal transduction, and cell-to-cell signaling. AB - Phytoestrogens are natural products from plants that function as estrogen agonists. Here we briefly review the biochemistry, distribution, and proposed roles of phytoestrogens in plants. A primary role appears to be in host-microbe interactions. For example, results from our laboratory suggest that genistein plays multiple roles in soybean defense, including the initiation of hypersensitive (apoptotic) cell death and the activation or potentiation of the defense competency of soybean cells. Recent results from our laboratory suggest that mammalian estrogens and estrogen agonists (including other phytoestrogens and fungal estrogen agonists) and certain specific peroxisome proliferators can also potentiate defense competency in soybean. These findings suggest that some overlap in nuclear receptor or other lipid/steroid signaling pathways may exist between plants and animals. PMID- 19281357 TI - Chemical synthesis of [(13)c]daidzein. AB - Pharmacokinetic and metabolic studies of phytoestrogens of the isoflavone class have been hampered by the lack of suitable stable-isotope-labeled analogs. A method for preparation of a [(13)C]-labeled analog of daidzein is described. [2 (13)C]Daidzein was synthesized by reaction of [(13)C]diethoxydimethylaminomethane with 2,4-dihydroxybenzoin. The final product was purified to more than 99% by reverse-phase high-performance liquid chromatography and structural analysis confirmed by nuclear magnetic resonance spectroscopy and mass spectrometry. Because [2-(13)C]daidzein is analytically and metabolically stable, it is a suitable analog for use as an internal standard for quantifying daidzein in biological fluids using isotope dilution mass spectrometry. This nonradioactive tracer is also ideal for investigating the pharmacokinetics of daidzein in humans because it is biologically indistinguishable from the unlabeled form. PMID- 19281358 TI - Synthesis of deuterated plant lignans for gas chromatography-mass spectrometry analysis. AB - Stabile deuterated plant lignans D(6)-matairesinol, D(8)-secoisolariciresinol, and D(8)-anhydrosecoisolariciresinol are synthesized to be used as internal standards in isotope dilution technique of gas chromatography-mass spectrometry in the selected ion monitoring mode. PMID- 19281359 TI - Phytoestrogens in foods and plants: extraction and isolation. PMID- 19281360 TI - Liquid chromatography: mass spectrometry of isoflavones. AB - High-performance liquid chromatography (HPLC) or capillary electrophoresis (CE) is particularly suited to the separation and analysis of a wide range of biological compounds. When investigating such compounds in complex matrices (e.g., blood, tissue, urine, foods), specificity of the detection method becomes an important issue. This can be overcome by extensive purification procedures before HPLC or CE analysis, but at the cost of a lot of effort and often unknown losses during the extraction and purification procedures. Such losses can be overcome by the addition of isotopically labeled internal standards, subject to their availability. The coupling of HPLC or CE with mass spectrometry provides a very specific method of detection. Compounds are transferred as ions to the gas phase by two types of spraying technologies, electrospray ionization (ESI) and heated nebulizer-atmospheric pressure chemical ionization. Although the mass-to charge ratios of the molecular ions can be used to provisionally identify a compound, full confirmation of structure requires collision-induced dissociation and analysis of the resulting fragment ions. Combination of specific parent ion daughter ion pairs allows for the quantitative measurement of isoflavones in the 1-5 pmol range with coefficients of variation for duplicate samples in the range of 5-9%. CE places greater demands on the mass spectrometer than does HPLC, because it generates narrow peak widths. A mass spectrometer with a scanning quadrupole analyzer does not enable full exploitation of the power of CE; in this context, an instrument with an ESI interface and a time-of-flight analyzer is ideal. PMID- 19281361 TI - Application of HPLC coupled with ultraviolet-photodiode array detection for the analysis of phytoestrogens in biological samples. AB - HPLC/PDA methodology was developed for phytoestrogen analysis in human blood, urine, and breast milk. Applications of this technique in four different observational studies showed that urinary excretion rates of isoflavonoids reflects soy consumption reliably. A case-control study in Shanghai showed a decrease in risk to develop breast cancer for women with high urinary isoflavone excretion. These findings support a potential breast cancer preventive effect achieved by soy consumption in populations that eat soy foods habitually. PMID- 19281362 TI - HPLC-electrochemical detection for phytoestrogen analysis: isoflavones and lignans in human serum and animal tissue. PMID- 19281363 TI - Immunoassay of phytoestrogens in human plasma. AB - Highly sensitive plasma immunoassay methods based on time-resolved fluorometry were developed for plasma enterolactone, genistein, and daidzein. For daidzein and genistein three types of methods and for enterolactone two different methods were developed and validated and the results compared with our gas chromatographic-mass spectrometric reference method. All three compounds may be determined in duplicate in a 2-300 mu plasma sample, even in subjects with low phytoestrogen values. PMID- 19281364 TI - Protein binding interactions: approaches to quantifying "free" and "bound" fractions of genistein and other xenobiotic estrogens in serum. PMID- 19281365 TI - Methods to screen estrogen-agonists and antagonists. PMID- 19281366 TI - Use of ERE and reporter gene constructs to assess putative estrogenic activity. AB - Estrogens primarily function through the activation of their receptors, which subsequently function as nuclear transcription factors. There are two estrogen receptor (ER) genes, now designated ERa (the classic ER gene) and ER/3. The key consequence of the activation of either gene product is the regulation of gene transcription. The extent and nature of transcription appear to be regulated by a series of coregulator proteins. One of the most sensitive assays for detection of potential estrogenic activity is measurement of the ability of a test compound to influence the transcription of reporter genes. In this regard, many investigators use promoter-reporter constructs. To assess putative estrogenic activity, an estrogen-responsive promoter is generally placed upstream of a reporter gene and transiently transfected into a target cell. When exposed to an estrogenic compound, expression of the reporter gene would normally be induced. We briefly discuss several issues pertinent to the use of these assays and the interpretation of resulting data, including estrogen-responsive, promoter reporter constructs, reporter genes and measurements of activity, choice of target cell or cell line, transient introduction of promoter-reporter constructs into cells, basic statistical approaches to data analysis, and definitions of agonist, partial agonist, and antagonist. PMID- 19281367 TI - Genistein: breast cancer protection and in vivo mechanisms of action. AB - We have hypothesized that early exposure to soy confers lifetime protection against breast cancer. This was investigated in Sprague Dawley CD rats exposed neonatally and prepubertally to injections of pharmacological doses of genistein and perinatally to physiological doses of genistein in the diet. These treatments resulted in a reduced number of dimethylbenz[a]anthracene (DMBA)-induced mammary tumors in the adults. The initial effect of early exposure to genistein was to upregulate the epidermal growth factor (EGF) signaling pathway and to downregulate transforming growth factor-beta (TGF-beta) expression in young animals. These actions enhanced cell differentiation and resulted in reduced EGF receptor expression in terminal end buds of adults. These differentiated cells are less proliferative and less susceptible to carcinogenesis. Bioavailability studies revealed that 25 and 250 mg genistein per kilogram of body weight diet resulted in total genistein concentrations of 54 and 1810 pmol/ml in serum of prepubertal rats. Genistein is demonstrated to be bioavailable to the mammary gland in postnatal rats. Prenatal genistein in the diet (250 mg/kg) did not protect against DMBA-induced mammary cancer or result in significant toxicity to the F(1) female reproductive tract. Prenatal exposure to genistein via the diet yielded only 43 pmol total genistein per milliliter of blood in the fetus. We conclude that in utero exposure to genistein via the diet does not protect against mammary cancer or cause toxicity because conjugated genistein cannot cross the placental barrier. Our results demonstrate in the rat that postnatal exposure to physiological concentrations of genistein in the diet is a safe and effective means of programming against chemically induced mammary cancer. PMID- 19281368 TI - The estrogen receptor-alpha knockout mouse and effects of phytoestrogens. PMID- 19281369 TI - Estrogen receptor beta and its interaction with phytoestrogens. PMID- 19281370 TI - Antioxidant properties of phytoestrogens. AB - The oxidation of lipids is an autocatalytic process consisting of a number of well-defined interrelated chemical reactions. Its importance has long been recognized in the food and polymer industry, and recent advances in the understanding of vascular diseases have shown that lipid peroxidation also contributes to human disease. The various chemical stages of the reaction offer several therapeutic targets for inhibition, and from the structural characteristics of phytoestrogens it is anticipated that they should exhibit antioxidant properties. Alone, it is not sufficient for compounds such as the phytoestrogens to exhibit biological activity as antioxidants; the criteria that should be satisfied for this mechanism to be relevant biologically are discussed. PMID- 19281371 TI - Application of topoisomerase assays in the evaluation of natural products as antitumor agents. AB - Initially, DNA topoisomerase (topo) inhibitors found clinical applications as antibiotics and cancer chemotherapeutic agents. Recently, we demonstrated that plant flavonoids that inhibit mammalian topo I or topo II might be useful as cancer chemopreventive agents (Constantinou et al., 1995b). Phytochemicals can inhibit DNA topoisomerases in different ways; depending on the mode and the type of enzyme, these can be classified as topo I poisons, topo II poisons, topo I antagonists, or topo II antagonists. Correctly classifying topo inhibitors is critical because it provides an important lead as to whether the plant agent can be useful in chemoprevention or in chemotherapy. We outline below a strategy that was designed to identify and classify topo I and II inhibitors. PMID- 19281372 TI - In vivo toxicity, pharmacokinetic features, and immunogenicity of B43(Anti-CD19) genistein immunoconjugate in mice and non-human primates. PMID- 19281373 TI - Multiple intracellular signaling pathways cross-talk with the estrogen receptor. PMID- 19281374 TI - Isoflavone-mediated inhibition of tyrosine kinase: a novel antiinflammatory approach. AB - Tyrosine kinase (TK)-mediated phosphorylation regulates signal transduction pathways resulting in the expression of a variety of inflammatory genes. Inhibition of TK activity in vivo has been shown to increase survival in a lethal model of murine endotoxemia, suggesting a novel therapeutic approach to inflammation and circulatory shock. We examined the role of TK activity on the expression of the inducible nitric oxide (NO) synthase (iNOS). Under resting conditions, iNOS is not expressed in human cells. In response to various proinflammatory stimuli, however, iNOS expression is upregulated, resulting in high-output NO synthesis. iNOS-derived NO plays a critical role as a cytotoxic effector species and has been implicated in the pathogenesis of many clinical inflammatory conditions, including inflammatory bowel disease, arthritis, transplant rejection, diabetes, and sepsis. We examined the signaling pathways governing iNOS expression in monolayers of DLD-1 cells, a human epithelial cell line derived from an intestinal adenocarcinoma. Induction of iNOS transcription in interferon-gamma-primed cells by treatment with lipopolysaccharide, Salmonella sp., or interleukin-1beta was potently inhibited by pretreatment with genistein, an isoflavone derived from the soy species genistin. Other isoflavones, such as genistin, daidzein, and daidzin, were not inhibitory. TK inhibition by genistein had no effect on the expression or nuclear translocation of the transcription factors interferon regulatory factor-1 and nuclear factor-KB, respectively, both of which have been implicated in transcriptional regulation of the human iNOS gene. Nuclear run-on analysis demonstrated that the effect of genistein on iNOS messenger RNA expression was not at the level of transcription, suggesting that posttranscriptional regulation of iNOS messenger RNA might be TK dependent. Isoflavones, such as genistein, are useful tools to dissect regulatory pathways in vitro and in vivo and may have potential use as novel antiinflammatory therapeutic agents. PMID- 19281375 TI - Analytical method for isoflavones in food. PMID- 19281376 TI - DNA markers associated with Loci underlying seed phytoestrogen content in soybeans. PMID- 19281377 TI - Isoflavone content in seeds of soybean cultivars grown in different areas of Brazil. PMID- 19281378 TI - Influence of variety, location, and growth year on phytoestrogen levels in ontario soybeans and processed soy food products. PMID- 19281379 TI - Method for measurement of dietary secoisolariciresinol using HPLC with multichannel electrochemical detection. AB - We describe a sensitive and specific assay for the determination of the plant lignan secoisolariciresinol, one of the main dietary precursors to the mammalian derived lignans, enterodiol and enterolactone. Quantification of secoisolariciresinol aglycone is achieved by reversephase high-performance liquid chromatography with multichannel electrochemical detection after hydrolysis of the glycoside moieties. This approach affords greater specificity than conventional ultraviolet detection and has a detection limit of 2.8 pmol. The method is ideally suited to the determination of secoisolariciresinol in processed flaxseed samples and can be used to assess the level of incorporation of flaxseed in fortified foods. PMID- 19281380 TI - Lignans and isoflavonoid polyphenols in tea and coffee. PMID- 19281381 TI - Development of a liquid chromatography-mass spectrometry procedure for quantitation of free and conjugated phytoestrogens in human urine: application in pharmacokinetic studies after soy consumption. PMID- 19281382 TI - Radioimmunoassay of phytoestrogens of isoflavone series. PMID- 19281383 TI - Method for the detection of nucleosides, bases, and hydroxylated adducts using gradient HPLC with coulometric array and ultraviolet detection. AB - Free radicals are compounds with an unpaired electron capable of independent existence. These highly reactive species have been implicated in many disease states and can react with cell membranes, lipids, proteins, and DNA. When an oxygen radical reacts with DNA, base damage, cross-linking (e.g., DNA-DNA or DNA protein), or DNA backbone damage (e.g., single- or double-strand breaks) can occur and often result in cell death. The field of oxidative metabolism as it relates to DNA damage has grown tremendously, with more DNA adducts being identified as biomarkers. These biomarkers are indicative of DNA damage. Measurement of these biomarkers has proved to be a challenge because of their relatively low occurrence (1 per 10(5)-10(6) bases). Methodologies for the measurement of DNA damage include thin-layer chromatography, enzyme-linked immunosorbent assay, gas chromatography-mass spectrometry, DNA sequencing, high performance liquid chromatography (HPLC)-ultraviolet, and HPLC-ECD. HPLC-ECD (electrochemical detection) is a powerful technique that is both sensitive and selective. However, HPLC-ECD is generally not amenable to gradient analyses, so its utility is restricted. In addition, many of the bases and nucleosides are not electrochemically active. Gradient HPLC separation coupled to both a coulometric electrochemical array detector and an ultraviolet detector overcomes these limitations. Presented here is a gradient HPLC method that measures a wide variety of nucleosides, bases, and hydroxylated adducts using the inherent stability, sensitivity, and wide dynamic range of a coulometric electrochemical array detector and the universal detection qualities of an ultraviolet detector. Linear ranges, limits of detection, and detailed methods development are presented. PMID- 19281384 TI - Assessing normal levels of phytoestrogens in a general population survey. PMID- 19281385 TI - Serum and urinary isoflavonoids and their metabolites in elderly men on diets supplemented with beverages containing untreated and alcohol-extracted soy protein. PMID- 19281386 TI - Development of a rat total enteral nutrition model for delivery of high levels of dietary phytoestrogens using soy protein isolate. PMID- 19281387 TI - In vitro and in vivo estrogenic activity of dietary flavonoids: importance of bioavailability and metabolism. PMID- 19281388 TI - Influence of human infant formulas containing phytoestrogens on brain aromatase activity in adult rats. AB - The biosynthesis of estrogens from androgen precursors is regulated by the aromatase enzyme, and estrogenic molecules are known to be significant factors in the formation of sexually dimorphic brain structures in the anterior hypothalamus. Phytoestrogens have been shown to inhibit aromatase enzyme activity in peripheral tissues. In this study, phytoestrogen-containing human infant formulas were examined to determine whether these estrogen mimics influence brain aromatase levels in adult rats. PMID- 19281389 TI - Inhibition of 17beta-Estradiol Formation by Isoflavonoids and Flavonoids in Cultured JEG-3 Cells: Search for Aromatase-Targeting Dietary Compounds. PMID- 19281390 TI - Effect of genistein and genistein analogs on growth of human prostate tissue in vitro. PMID- 19281391 TI - Inhibition of growth of primary human prostate cells by phytoestrogens. PMID- 19281392 TI - Dietary soy protein and soy isoflavones: histological examination of reproductive tissues in female rats. PMID- 19281393 TI - Antioxidant potential of phytoestrogens. PMID- 19281394 TI - Antioxidant intracellular activity of genistein and equol. AB - Antioxidant effects of isoflavonoids have recently been described. To learn whether the isoflavonoids genistein and equol have actions on the intracellular free radicals, human neutrophils and J774 monocyte-macrophage cell line were used to measure the intracellular production of O(2) (superoxide anion) and H(2)O(2) (hydrogen peroxide) by flow cytometry. The results shown significatives decrease in O(2) and H(2)O(2) production after 1 hour of incubation with equol and genistein. The phagcytic oxidant production decreased owing to the effects of both isoflavonoids in a concentration-dependent manner. PMID- 19281395 TI - Natriuretic effect of equol. AB - A furosemide-sensitive Na-K-Cl cotransporter (NKCC2 isoform) accounts for almost all luminal NaCl reabsorption in the thick ascending limb of Henle's loop (TALH). The activity of this transport protein is regulated by humoral factors known as cotransport inhibitory factors. One family of these compounds is represented by the urinary phytoestrogens equol and genistein, which inhibit cotransport fluxes at concentrations similar to furosemide. Moreover, they possess salidiuretic potency similar to furosemide in the isolated perfused rat kidney, but are less potent than furosemide (in vivo). Thus, dietary phytoestrogens can be responsible, at least in part, for the low blood pressure of vegetarians. PMID- 19281396 TI - Effects of soy protein and isoflavones on plasma lipid profiles in patients with cardiovascular disease. PMID- 19281397 TI - Association between plasma isoflavone and plasma lipoprotein concentrations. PMID- 19281398 TI - Phytoestrogen and intact soy protein consumption differ in their effects on plasma lipids and lipoproteins. PMID- 19281399 TI - Effects of soy protein and soy phytoestrogens on symptoms associated with cardiovascular disease in rats. PMID- 19281400 TI - Government-funded phytoestrogen research in the United kingdom. PMID- 19281402 TI - Response to letter to the editor. PMID- 19281401 TI - Strain-independent variation of phytoestrogen content in soybeans mitigates significance of variation in genetically modified varieties. PMID- 19281403 TI - Newswatch. PMID- 19281406 TI - Antioxidant capacity of edible plants: extraction protocol and direct evaluation by cyclic voltammetry. AB - Reactive oxygen-derived species are produced in cells under physiological conditions and in response to stress. Among the various antioxidant systems responsible for protection against these species, the low-molecular-weight antioxidants (LMWA), such as ascorbate, play an important role. Cyclic voltammetry (CV) has been proposed as a tool for quantitation of the total antioxidant capacity of plasma. It has also been shown that biological oxidation potentials, as determined from the anodic current waves of the CV tracings, are specific characteristics of the various LMWA components, and that the amplitude of each wave can be used for quantitation of the specific component. The adaptation of CV for evaluation of the total antioxidant capacity of edible plants is demonstrated here. The area under the anodic current wave is proposed as a better indicator for the content of LMWA, compared with the amplitude. This distinction could prove valuable when more than a single molecule contributes toward a specific anodic wave and when the identities of the components of a wave are not known. Vegetables and fruits that are commonly consumed in the U.S. diet were used. They were extracted with either water, aqueous acetic acid (30%), or a mixture of water, acetic acid, and acetonitrile (40:30:30). The LMWA contents were evaluated by CV. In three to five steps the LMWAs were completely extracted from the edible foods, and their amounts were translated into equivalents of ascorbate. PMID- 19281407 TI - Marketing Functional Foods: What Have We Learned? An Examination of the Metamucil, Benefit, and Heartwise Introductions as Cholesterol-Reducing Ready-to Eat Cereals. AB - Health claims, U.S. Food and Drug Administration regulatory actions, and marketing practices are examined in the environment precipitating the introduction of the Nutrition and Labeling Education Act (NLEA) in 1990. The introduction of three psyllium-based products in 1989 and 1990, Procter & Gamble's Metamucil wafers, General Mills' Benefit cereal, and Kellogg's Heartwise cereal, are examined for their use of health claims and marketing tactics in a changing regulatory environment. Inconsistent approval of psyllium-based health claims for drug and food products created a confusing environment in the pre- and early post-NLEA era. This was exacerbated by marketing issues regarding consumer communication and product positioning. Suggestions for marketing products with health positionings in the new NLEA environment are proposed. PMID- 19281408 TI - Determination of the Fatty Acid content of pumpkin seed, pygeum, and saw palmetto. AB - Fatty acids are major components of many plants, foods and medicines, including pumpkin seeds (Cucubita pepo), pygeum bark (Prunus africana) and saw palmetto (Serenoa repens). With the gas chromatography methods reported here, free fatty acids of these species can be quantified as their trimethylsilyl derivatives. Because of their different fatty acid contents and composition, the gas chromatography method can distinguish which of three plant species was extracted, and, in the case of S. repens, the method of extraction. Although phytosterols can be separated by this method, their content is too low to be assigned directly. The total fatty acid content can be determined through formation of the methyl esters. This is helpful for estimation of the kind and percentage of fatty acids that are present as triglyceride esters in the plant material and for standardization of the products. PMID- 19281409 TI - My folate valentine. PMID- 19281410 TI - Newswatch. PMID- 19281412 TI - Characterization of a novel relapsing fever spirochete in the midgut, coxal fluid, and salivary glands of the bat tick Carios kelleyi. AB - Bat ticks, Carios kelleyi, from Iowa were examined for the presence of relapsing fever group borreliae. A novel spirochete was characterized by DNA sequence analysis of polymerase chain reaction amplicons for the 16S rRNA, flaB, and glpQ genes in either triturated tick pools or single ticks. All loci and the concatenated DNA sequence of 3,289 bases identified the Carios bacterium as a relapsing fever spirochete most closely related to, but distinct from, Borrelia turicatae. Spirochetes reactive with a Borrelia-specific monoclonal antibody were observed microscopically in the coxal fluid and salivary glands from one tick. These data confirm the presence of a novel species of relapsing fever spirochete in bat ticks and the potential for new enzootic foci for endemic relapsing fever that warrants further investigation. The name Borrelia johnsonii is proposed for this novel spirochete in honor of Dr. Russell C. Johnson. PMID- 19281413 TI - Clinical evaluation of Nd:YAG and 685-nm diode laser therapy for desensitization of teeth with gingival recession. AB - OBJECTIVES: The aim of this study was to evaluate the effectiveness of two types of lasers, the Nd:YAG laser and the 685-nm diode laser, as dentin desensitizers as well as both the immediate and late therapeutic effects on teeth with gingival recession. MATERIALS AND METHODS: The study was conducted on 56 teeth in 14 patients with Miller's class 1 and 2 gingival recession with clinically elicitable dentin hypersensitivity (DH). The patients were divided into two groups: a Nd:YAG-laser-treated group and a 685-nm diode laser-treated group. DH was assessed by means of an air stimulus, and a visual analog scale (VAS) was used to measure DH. The selected teeth in the two groups received laser therapy for three sessions. Teeth subjected to Nd:YAG-laser treatment were irradiated at 1 W and 10 Hz for 60 sec at 1064 nm, and those receiving 685-nm diode laser treatment were irradiated at 25 mW and 9 Hz for 100 sec. RESULTS: Significant reductions in DH occurred at all time points measured during the three treatment sessions in both treatment groups. Comparing the means of the responses in the three treatment sessions for the two groups revealed that the Nd:YAG laser group had a higher degree of desensitization compared to the other group (p<0.01). The immediate and late therapeutic effects of the Nd:YAG laser were more evident than those of the 685-nm diode laser. CONCLUSIONS: Both of these lasers can be used to reduce DH without adverse effects. Desensitization of teeth with gingival recession with the Nd:YAG laser was more effective than with the diode laser. The Nd:YAG laser appears to be a promising new tool for successfully reducing DH. PMID- 19281414 TI - Cerebral autoregulatory response depends on the direction of change in perfusion pressure. AB - The purpose of cerebral autoregulation is to keep cerebral blood flow constant during variations of cerebral perfusion pressure (CPP). Recently, the autoregulatory response was reported to be greater during arterial blood pressure (ABP) increase than during decrease following repeated induced changes in ABP in 14 brain-injured subjects. The goal of this study was to further investigate the asymmetry of autoregulation during spontaneous increases and decreases of CPP in a larger group of brain injury patients. Data recordings (N=727) of CPP and cerebral blood flow velocity (CBFV) in 210 subjects with traumatic brain injury (TBI) were studied. Autoregulation was assessed using moving correlation indices (Mx) between CPP and CBFV. Periods of increasing and decreasing CPP were separately correlated to corresponding CBFV in order to assess autoregulatory responses to upward (upMx) and downward (downMx) changes of CPP. These correlation indices range from -1 to +1; negative or zero values indicate intact autoregulation, whereas positive values indicate impaired autoregulation. Only data with defined strong CPP variations were evaluated. Strong CPP variations were found in 84 recordings of 53 patients. On average (+/-SD) upMx was significantly lower than downMx (0.05+/-0.49 versus 0.14+/-0.54; p < 0.005). Despite this difference, upMx and downMx were strongly correlated with each other (R=0.82; p < 0.001). In conclusion, the autoregulatory response was significantly greater during increase than during decrease in CPP. The results may indicate non linear behavior of cerebral autoregulation. PMID- 19281415 TI - Neonatal encephalopathy: treatment with hypothermia. AB - In this article, the role of hypothermia and neuroprotection for neonatal encephalopathy will be discussed. The incidence of encephalopathy due to hypoxia ischemia as well as the pathophysiology will be presented. The diagnosis of encephalopathy in full-term neonates will be discussed. The current management of brain injury that occurs with hypoxia ischemia and the role of hypothermia in preventing brain injury in fetal and neonatal animal models will be reviewed. The current data from randomized control trials of hypothermia as neuroprotection for full-term infants will be presented along with the results of meta-analyses of these trials. Lastly, the status of ongoing neonatal hypothermia trials will be summarized. PMID- 19281416 TI - Should intraoperative hypercapnea or hypercarbia raise concern in neonates undergoing thoracoscopic repair of diaphragmatic hernia of Bochdalek? AB - BACKGROUND: Better understanding of cardiopulmonary physiology in children with congenital diaphragmatic hernia of Bochdalek (CDH) has facilitated improved survival. In addition, it has allowed surgeons to employ minimally invasive techniques to their repair under conditions that might result in hypercarbia and/or recurrent pulmonary hypertension. MATERIALS AND METHODS: Under institutional review board approval, the records of all neonates at a single institution who underwent thoracoscopic CDH (T-CDH) were reviewed with attention to the incidence of intraoperative hypercapnea (elevated end tidal CO(2)) or hypercarbia (increased PCO(2) by blood gas measurement) and any associated complications. RESULTS: From 2004 through 2007, 31 consecutive neonates, including those who had undergone extracorporeal membrane oxygenation, had T-CDH. CO(2) insufflation to 3 mm Hg was used until the viscera were reduced within the abdominal cavity. The operative technique and outcomes are described elsewhere. Preoperative analyses revealed a mean arterial PCO(2) of 53 +/- 11 torr and SaO(2) of 95 +/- 5%. The mean highest recorded intraoperative end-tidal CO(2) level was 64 +/- 13 and correlated poorly with the highest arterial PCO(2) (mean, 78 +/- 29 torr; range, 29-130). The mean lowest recorded intraoperative SaO(2) was 92 +/- 8% with only two values less than 88%. The average lowest intraoperative mean arterial blood pressure was 47 +/- 8 mm Hg (range, 34-70 mm Hg). No neonate received inhaled nitric oxide, intravenous buffer administration, or escalation of inotrope administration during the procedures nor did any experience recurrent pulmonary hypertension postoperatively. SUMMARY: Hypercapnea and hypercarbia are common phenomena during T-CDH but do not appear to correlate with one another nor result in clinically evident recurrent pulmonary hypertension, hypoxemia, hypotension, need for support with vasoactive medications, inhaled nitric oxide, or buffering agents. PMID- 19281417 TI - Laparoscopic gastropexy for the treatment of gastric volvulus associated with wandering spleen. AB - A 2.5-year-old boy was referred to the emergency room for a sudden onset of diffuse and increasing abdominal pain with lethargy, abdominal distension, and vomiting, all in the past 24 hours. A plain abdominal X-ray showed gastric distension. Two liters of gastric contents were evacuated by suction. The abdominal sonogram showed an unusual position of the spleen in the left-lower quadrant, with no splenic ischemia. The diagnosis of gastric volvulus associated with a wandering spleen was then evoked. Laparoscopic exploration revealed a nonischemic spleen, absence of normal supporting ligaments for the spleen, and gastric distension with flaccid gastric walls. The spleen was then easily moved in the left-under quadrant. A parietal peritoneal posterolateral incision was made, opposite the large gastric curve, up to the diaphragm (7 cm). This delimitated a sharp demarcation zone between the two edges of the incised peritoneum. The stomach was fixed to the peritoneal incision, covering and anchoring the spleen in a good position. Recovery was uneventful, and an abdominal sonogram performed 4 years after the surgery shows a viable spleen in its correct location. The rarity of gastric volvulus associated with a wandering spleen and its fast clinical improvement with medical treatment often delays the diagnosis and the surgical treatment. Laparoscopy in this case has a dual relevance: diagnosis and therapeutic management (splenectomy or gastropexy). Laparoscopic gastropexy for the treatment of gastric volvulus associated with a wandering spleen is an easy procedure and combines the advantages of all the surgical techniques previously described. PMID- 19281418 TI - Technique of laparoscopic transperitoneal lower pole heminephroureterectomy. AB - BACKGROUND: Laparoscopic transperitoneal nephrectomy is technically feasible in most cases of benign renal disease. To date, there have been isolated reports of laparoscopic transperitoneal heminephroureterectomy of principally the upper moiety of a duplex system. Rarely reported are lower moiety nephroureterectomies. AIM: The aim of this IPEG submission is to present a 3-minute video demonstrating the technical details of a transperitoneal laparoscopic, right, lower moiety, heminephroureterectomy in a 3-year-old. METHODS/TECHNIQUES: A child with marginally functioning (<5%) lower moiety of a right duplex system was subjected to a transperitoneal heminephroureterectomy. The patient was placed in a left lateral position and an umbilical camera and two working ports (right iliac fossa, epigastric), all 5 mm, were utilized. The ascending colon was reflected to the left and the underlying lower moiety ureter identified and isolated. With traction on the ureter, the pelvis was brought into view and control of blood supply to the lower moiety was achieved by using a combination of ultrasonic scalpel and hook diathermy. A critical step at this stage was definition of the superior limit of the pelvis, which corresponded to the level of vascular demarcation. The renal parenchyma was transected at this point by using the ultrasonic scalpel, which ensured reasonable hemostasis. Complete hemostasis was assured by approximating the divided kidney with a series of three interrupted intracorporeal sutures. The subtending ureter was dissected close to the bladder base, where it was ligated and divided. A drain was placed percutaneously in the renal bed and the specimen retrieved via an extended umbilical incision. RESULTS: The patient was started on fluids on recovery with oral feeds introduced the next morning. The drain was removed prior to discharge on day 2. At 6 months post resection, the patient remains well. CONCLUSION: Laparoscopic transperitoneal lower pole heminephroureterectomy is technically feasible for benign renal disease in children. The combination of ultrasonic scalpel and intracorporeal suturing is adequate to control bleeding of the transected kidney. Dissection of the ureter distally to bladder neck is easily achieved without change in port position. Recovery is robust with minimal requirement for analgesia. The wider space, better view, and ability to access all of the urinary tract make this a tenable alternative to other laparoscopic approaches. PMID- 19281419 TI - Long-term functional evaluation of fecal continence after laparoscopic-assisted pull-through for high anorectal malformations. AB - BACKGROUND: The use of laparoscopy while performing an abdominal dissection for the mobilization of rectovesical fistula should have an impact on anorectal function, as compared to the original posterior sagittal anorectoplasty, where muscle complex was not cut and the rectum was pulled in a way similar to the old techniques for the abdominoperineal pull-through. This necessitates a functional reevaluation. MATERIALS AND METHODS: A prospective case study included 15 patients treated with a laparoscopic-assisted pull-through for high anorectal malformation. Laparoscopy was used for abdominal dissection and ligation of the fistula, with the pull-through completed by a small perineal incision centered over the external sphincter. Nine of them are now older than 3 years for fecal continence evaluation. After the approval of the Ethical Committee for Medical Research in the Department of Surgery at Ain Sams University (Cairo, Egypt) and obtaining an informed consent from the parents, they were subjected to a full clinical history and a checklist about motions and soiling to be filled in over 1 month, a barium enema to check for any dilatation, anorectal manometry to evaluate resting pressure, maximum squeezing pressure, and sphincter relaxation, and MRI (magnetic resonance imaging) to evaluate the central situation of the rectum within the sphincter and the degree of development of the sphincter. Their degree of continence was graded according to the Kelly score. RESULTS: Six of 9 patients are clean without any attacks of fecal soiling or incontinence, and they evacuate spontaneously but need the application of a rectal suppository for evacuation from time to time. The remaining 3 patients had variable degrees of fecal incontinence. One of them had mucosal prolapse and was excised with good cleanliness postoperatively. The remaining 2 patients are managed by medical control and they are clean with minimal soiling when stools are loose. MRI and barium enema showed a centrally placed rectum in the muscle complex without dilatation in all cases. Manometry showed a high resting pressure that decreased on straining in the 7 clean patients and low in 2. The resting pressure did not increase on squeezing and all showed weak rectoanal inhibitory reflex (RAIR). One patient developed dysurea and constipation 1 year after surgery, as diagnosed by VCUG (voiding cystourethrogram) to have a diverticulum at the site of excised fistula causing rectal and urethral obstruction treated by a transabdominal excision with a good functional result. CONCLUSION: The state of continence with the laparoscopic technique in high anorectal malformations in this study showed acceptable results but needs bigger series with longer follow-up for a proper evaluation of this technique. PMID- 19281420 TI - Effect of elevated intra-abdominal pressure on portal vein and superior mesenteric artery blood flow in a rat. AB - AIM: Recent clinical experience suggests that minimal access portoenterostomy (the Kasai procedure) for biliary atresia leads to transplantation sooner, compared to the traditional open approach. It should be emphasized that elevated intra-abdominal pressure (IAP) may reduce hepatic and portal blood flow and thus may cause histologic liver damage. The aim of the present study was to evaluate the effects of IAP on blood flow in the portal vein (PV), compared to the superior mesenteric artery (SMA), and on the systemic mean arterial blood pressure (MABP). MATERIALS AND METHODS: Male Sprague-Dawley rats were anesthetized with intraperitoneal ketamine (90 mg per kg) and xylasine (13 mg per kg). Polyethylene catheters (PE-50) were introduced into the right carotid artery for the measurement of MABP. After a midline laparotomy, the SMA and PV were isolated. Ultrasonic blood-flow probes were placed on the vessels for the continuous measurement of regional blood flow. Two large-caliber percutaneous peripheral intravenous catheters were introduced into the peritoneal cavity for inflation of air and for the measurement of IAP. The time course of MABP and SMA and PV flow as well as the relationship between IAP and SMA and PV flow were determined. RESULTS: Although all three hemodynamic parameters decreased with the increase in the IAP, the most significant changes were observed in PV blood flow. IAP at 3 mm Hg resulted in a 26% decrease in PV flow (P < 0.05), a 19% decrease in SMA flow (P < 0.05), and an 11% decrease in MABP (P < 0.05). IAP at 6 mm Hg caused a two-fold decrease in PV flow (P < 0.05), a 30% decrease in SMA flow (P < 0.05), and a 19% decrease in MABP (P < 0.05). There were no changes in the time course of MABP and PV and SMA flow. PV and SMA flow returned to normal values immediately after abdominal deflation. CONCLUSIONS: Persistent IAP decreased MABP, SMA, and, especially, PV flow by 50%. We speculate that in biliary atresia patients with already present liver dysfunction, decrease in SMA flow and even a greater decrease in PV flow from increased IAP, which occurs during a laparoscopic Kasai procedure, may further compromise liver function. This may be one of the explanations for the progression to earlier transplantation in infants undergoing a laparoscopic Kasai procedure. PMID- 19281421 TI - Review of laparoscopic training in pediatric surgery in the United Kingdom. AB - AIM: To review the exposure pediatric surgery trainees have to laparoscopic surgery in the United Kingdom (UK). METHODS: A confidential postal questionnaire was sent to all trainees working at registrar level in centers responsible for pediatric surgical training in the UK. Questions assessed the number of consultants with an interest in laparoscopic surgery, types of cases performed laparoscopically, and trainees' role in laparoscopic appendicectomy (LA). RESULTS: Questionnaires were sent to 112 trainees with a 55% response rate (62 replies). At least one response was received from each unit. Based on responses, 49 to 67 consultants in 21 training centers have an interest in laparoscopic surgery (0%-100% of consultants per unit). LA was offered in 20 out of 21 training centers. There was no significant difference in the proportion of appendicectomies performed laparoscopically by junior (years 1-3) and senior (years 4-6) trainees. A significantly higher proportion of junior trainees had not performed any LAs (P = 0.02). Seventy-three percent of trainees were the principal operator. For trainees who were principal operators, the cameraperson was a consultant in 52% and a junior trainee in 17%. The time of day affected the likelihood of a procedure being carried out laparoscopically in 43 (81%) responses. CONCLUSIONS: The majority of trainees' exposure to laparoscopic surgery could be viewed as suboptimal; however, the exposure gained varies significantly between different units throughout the UK. In an age moving in favor of minimal access surgery, all units must be in a position to offer pediatric laparoscopic surgical training. PMID- 19281422 TI - Laparoscopic treatment of pancreatic pseudocysts in children. AB - BACKGROUND: Pancreatic pseudocysts are problematic sequelae of pancreatitis or pancreatic trauma causing persistent abdominal pain, nausea, and gastric outlet obstruction. Due to the low volume of disease in children, there is scant information in the literature on the operative management of pseudocysts with minimally invasive techniques. We conducted a multi-institutional review to illustrate several technical variations utilized in achieving laparoscopic cystgastrostomy in the pediatric population. METHODS: A retrospective review was conducted of all patients who underwent laparoscopic cystgastrostomy in five institutions. Patient data, operative techniques, and postoperative course were analyzed. RESULTS: There were 13 patients with a mean age of 10.4 years and mean weight of 52.1 kg. The etiologies of pancreatitis included: trauma (4), gallstones (3), chemotherapy (2), hereditary (1), and idiopathic (3). Preoperative radiographic measurements of the maximal cyst diameter averaged 11.7 cm. Cystgastrostomy was approached by using transgastric exposure in 5 cases and intragastric ports in 8 cases. An average of four ports were used to complete these operations. Mean operative time was 113 minutes. There were no conversions in this series. Cystgastrostomy was performed by using an endoscopic stapler (average 3.8 loads) in 6 cases, sutures in 6 cases, and 1 was formed solely with the Harmonic Scalpel (Johnson and Johnson). Gastrotomy sites were closed by using a stapler in 4 cases and suture techniques in 9. Mean time to initial and goal feeds was 3 and 4 days, respectively. Postoperative imaging revealed persistent pseudocyst in 1 patient, who was treated with a distal pancreatectomy. Therefore, 92% required no further operative intervention and remained asymptomatic upon recovery from their pancreatitis. CONCLUSION: A laparoscopic approach to pancreatic cystgastrostomy for chronic pseudocyst proved to be safe and effective in this five-institution survey. Techniques varied, but 92% had complete resolution with minimal morbidity and rapid recovery. Laparoscopic cystgastrostomy should be considered as an appropriate first-line treatment for chronic pseudocysts in children. PMID- 19281423 TI - Endoscopic repair of late-presenting Morgagni and Bochdalek hernia in children: case report and review of the literature. AB - BACKGROUND: Delayed presentation of a congenital diaphragmatic hernia (CDH) is not usual. Primary laparoscopic repair is becoming the standard in many centers. Different approaches and techniques have been proposed. There is not enough evidence in the literature to recommend one technique over another. PATIENTS AND METHODS: In this paper, we report one case of Morgagni hernia (MH) and one case of Bochdalek hernia (BH), 2 and 6 years old, respectively, operated on in our hospital. In the first case, the diaphragmatic defect was directly sutured with extracorporeal interrupted nonabsorbable sutures, passed through the abdominal wall with a Reverdin needle. In the second one, intracorporeal nonabsorbable stitches were placed. A search of the literature was made using PubMed and the following words: congenital diaphragmatic hernia, laparoscopy or thoracoscopy, and children. The neonatal Bochdalek hernias were discarded. Data about surgical approach, suturing technique, management of the hernia sac, complications, and recurrence were summarized for both pathologies. RESULTS: Both patients were discharged 48 hours after surgery. There were no complications. No recurrence was evident after 6 months. Eleven articles on the treatment of MHs were found, with a total of 30 patients. For BHs, 10 articles met the inclusion criteria, with a total of 54 patients. In both groups, all the papers were case reports or retrospective reviews of case series. The MH is best approached through laparoscopy, and the BH can be treated through thoracoscopy or laparoscopy. Most researchers prefer direct suture with nonabsorbable material. Both complication and recurrence rates are very low. CONCLUSIONS: The endoscopic approach of late presenting Morgagni and Bochdalek CDH is a safe technique. It offers all the advantages of minimally invasive surgery (MIS), and laparoscopy also permits the diagnosis and treatment of other associated conditions. There are a short number of cases reported and no prospective study comparing open with the MIS approach for the treatment of diaphragmatic malformations. PMID- 19281425 TI - Stress response and the value of reproduction: are birds prudent parents? AB - In vertebrates, stressors such as starvation or predator attacks stimulate the rapid elevation of circulating glucocorticoid hormones, triggering physiological and behavioral responses that aid immediate survival but simultaneously inhibit reproduction. This stress response has been proposed to serve as a physiological mediator of life-history trade-offs: when the value of current reproduction is high relative to the value of future reproduction and survival, a mitigated stress response is expected to enable successful breeding and maximize fitness. Using phylogenetic comparative analyses, we investigated baseline and peak stress induced plasma corticosterone levels during parental care in 64 bird species. We found that (1) species with a higher value of the current brood relative to future breeding mounted weaker corticosterone responses during acute stress, and (2) females in species with more female-biased parental care had weaker corticosterone responses. These results support the brood value hypothesis, suggesting that the stress response evolves as an adaptive basis for life-history strategies. Further, we found that (3) baseline corticosterone correlated positively with brood value and negatively with body mass, and (4) peak corticosterone was greater in species breeding at higher latitudes. The latter findings suggest that circulating corticosterone concentrations might be matched to the anticipated demands and risks during nesting. PMID- 19281426 TI - Once-daily insulin detemir in a cohort of insulin-naive patients with type 2 diabetes: a sub-analysis from the PREDICTIVE study. AB - OBJECTIVE: PREDICTIVE is a large, observational study of the empirical use of insulin detemir in patients with type 1 or type 2 diabetes (T1DM/T2DM). This post hoc analysis evaluates insulin-naive patients with T2DM uncontrolled on oral antidiabetic drugs (OADs) who were initiated and remained on once-daily insulin detemir for 12 weeks. RESEARCH DESIGN AND METHODS: This observational, multinational, multi-center, open-label prospective study evaluated the efficacy and safety of insulin detemir in 1653 insulin-naive patients with T2DM (mean age 60.8 +/- 10.9 years, BMI 29.8 +/- 4.8 kg/m(2), and HbA(1C) 8.82 +/- 1.50%). Statistical comparisons were made between baseline and 12-week follow up data. Our study was subject to the usual limitations of observational studies. MAIN OUTCOME MEASURES: Endpoints were: incidence of serious adverse drug reactions, including number of hypoglycemic events (total, major, and nocturnal), glycemic parameters, and weight change. RESULTS: Following insulin initiation, no significant change occurred in the number of nocturnal hypoglycemic events or total hypoglycemic events (p = 0.4513), and no serious adverse drug reactions were observed during the 12 weeks of treatment. HbA(1C) decreased by a mean 1.25% (SD +/- 1.25%; p < 0.0001), with 30% of patients (n = 383) achieving HbA(1C) <7% at 12 weeks. Mean changes in fasting blood glucose and fasting blood glucose variability were -3.62 mmol/L (SD +/- 2.93; p < 0.0001) and -0.48 mmol/L (SD +/- 1.03; p < 0.0001), respectively. Body weight decreased by a mean 0.5 kg (SD +/- 3.3; p < 0.0001), with weight loss or no weight change occurring in a substantial percentage of patients in each BMI category (<25, 25-30, 30-35, and >35 kg/m(2)). Patients with higher baseline BMI lost the most weight, with the greatest weight loss (-1.20 kg) reported in those with BMI >35 kg/m(2). CONCLUSIONS: Empirical use of insulin detemir as an insulin initiation strategy can improve glycemic control with good tolerability, including a low risk of hypoglycemia and a weight benefit, in a majority of insulin-naive patients uncontrolled on OADs. PMID- 19281427 TI - Anti-tumoral effect of a celecoxib low dose on a model of human medullary thyroid cancer in nude mice. AB - BACKGROUND: Medullary thyroid carcinoma (MTC) is a C cell neoplasm secreting calcitonin (CT). Surgery remains the only treatment as MTC is resistant to radio- and chemotherapies. Anti-tumoral effects of nonsteroidal anti-inflammatory drugs have been observed in various cancers. Thus, we tested the anti-tumoral action of an nonsteroidal anti-inflammatory drug, celecoxib, on MTC development. METHODS: We studied the expression of prostaglandin (PG) metabolism enzymes in our in vitro (TT cells) and in vivo (TT tumors) models and in human MTCs by Western blot. We checked the effect of celecoxib on xenografted subcutaneous tumors in nude mice. Celecoxib was administrated in powder food during 9 weeks from day 1 after TT cell injection. At the end of the experiment plasma CT was measured by radioimmunoassay, the number of proliferating cells in tumor tissues was detected by Ki67 immunocytochemistry and apoptotic nuclei by caspase 3 ad Bcl-2 expression and terminal-deoxynucleotidyl-transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) method. PGE(2) concentrations in TT cell medium were evaluated by an enzyme immunoassay kit. RESULTS: Our in vitro and in vivo models were validated: the status of PG metabolism enzymes was comparable in these models and in human MTCs. A very low dose of celecoxib, 120 ppm in food, inhibited tumor volume by 71% and reduced plasma CT level. Although no proapoptotic effect was detectable in tumors, a decrease of proliferating cells was revealed. The inducible PG synthesis enzyme, cyclooxygenase 2, was only detectable in rare stromal cells. The expression of the constitutive PG synthesis enzyme, cyclooxygenase 1, was diminished, while the level of the catabolism enzyme, 15-PG dehydrogenase, was decreased. In vitro, TT cells treated for 12 days with 25 muM celecoxib reproduced these changes, and PGE(2) secretion was not significantly modified by the treatment, in these conditions. CONCLUSION: Celecoxib has a good therapeutic potential for MTC to prevent metastasis growth, and its anti-tumoral effect is, at least in part, independent of PGE(2). PMID- 19281428 TI - The utility of radioiodine scans prior to iodine 131 ablation in patients with well-differentiated thyroid cancer. AB - BACKGROUND: The utility of radioiodine (RAI) scans prior to (131)I ablation is controversial. The objective of this study was to evaluate the utility of RAI scans prior to (131)I ablation in patient with well-differentiated thyroid cancer. METHOD: All RAI scans performed prior to (131)I ablation from July 2000 to November 2006 at Washington Hospital Center were reviewed retrospectively. Patients were excluded who were suspected of having 1) loco-regional disease, 2) distant metastases, and/or 3) physiological uptake that might alter management prior to the pre-ablation RAI scans. RAI scans were performed either 24 hours after dosing with 37-148 MBq of (123)I or 48 hours after dosing with 37-148 MBq of (131)I with imaging of the whole body, the thyroid bed/neck with a pinhole collimator, and the neck and chest with a parallel-hole collimator. One reviewer blindly evaluated each set of scans using six criteria, and for the purpose of this study, the thresholds for each criterion for which the patient's management may have been altered prior to (131)I ablation are noted in parentheses: 1) the number of foci of RAI uptake in thyroid bed/neck (0 or > or =6), 2) the location(s) of these foci in the thyroid bed/neck (outside the thyroid bed), 3) the size of the largest foci in thyroid bed/neck (> or =1 lobe), 4) the percent uptake in the thyroid bed/neck (> or =15%), 5) uptake suggestive of distant metastases, and 6) significant altered biodistribution (e.g., any breast, marked salivary gland, or marked gastrointestinal uptake). RESULTS: Of 355 sets of scans reviewed, 53% of patients had findings on the RAI scans that might have altered the patient's management prior to their (131)I ablation. The data grouped by the criteria noted above were 1) 12% with six or more foci suggesting local metastases and 6% (22) with no focal uptake, 2) 14% with suggestion of lymph node metastases, 3) 1.1% with at least one focus > or =1 lobe, 4) 8% with > or =15% uptake, 5) 4% with distant metastases, 6) 16% demonstrating altered distribution with 6% breast, 3% salivary, 10% GI, and 0.3% urinary bladder. CONCLUSION: Pre ablation RAI scans demonstrate a significant number of findings that may alter the management of patients with well-differentiated thyroid cancer prior to (131)I ablation. PMID- 19281429 TI - Second primary malignancy risk after radioactive iodine treatment for thyroid cancer: a systematic review and meta-analysis. AB - BACKGROUND: The risk of second primary malignancies (SPMs) associated with cancer therapies is an important concern of thyroid cancer survivors and physicians. Our objective was to determine if the risk of SPMs is increased in individuals with thyroid cancer treated with radioactive iodine (RAI), compared to those not treated with RAI. METHODS: We performed a systematic review of the literature and meta-analysis. Two independent reviewers screened citations and reviewed full text papers. If not reported by the primary authors, the relative risk (RR) of SPMs was calculated by dividing the standardized incidence ratio of SPM in individuals with thyroid cancer treated with RAI compared to those not treated with RAI (with associated 95% confidence intervals [CI]). The natural logarithms of RRs of respective SPMs, weighted by the inverse of the variance, were pooled using fixed effects models and the exponential of the results was reported. RESULTS: Two multi-center studies (one from Europe and the other from North America) were included in this review. The RR of SPMs in thyroid cancer survivors treated with RAI was significantly increased at 1.19 (95% confidence interval [CI] 1.04, 1.36, p = 0.010), relative to thyroid cancer survivors not treated with RAI (data from 16,502 individuals), using a minimum latency period of 2 to 3 years after thyroid cancer diagnosis. The RR of leukemia was also significantly increased in thyroid cancer survivors treated with RAI, with an RR of 2.5 (95% CI 1.13, 5.53, p = 0.024). We did not observe a significantly increased risk of the following cancers related to prior RAI treatment: bladder, breast, central nervous system, colon and rectum, digestive tract, stomach, pancreas, kidney (and renal pelvis), lung, or melanoma of skin. CONCLUSIONS: The risk of SPMs in thyroid cancer survivors treated with RAI is slightly increased compared to thyroid cancer survivors not treated with RAI. PMID- 19281430 TI - Cardiac diastolic dysfunction is prevalent in HIV-infected patients. AB - Combination antiretroviral therapy (ART) has markedly improved survival in HIV infected patients, but not without significant adverse effects including ART associated dyslipdemia and insulin resistance, which may in part contribute to an increased risk of cardiovascular events. Other contributing factors to cardiovascular risk may include uncontrolled HIV replication, the effects of HIV and ART on vascular endothelium and inflammatory cytokines. Diastolic dysfunction may be an early sign of cardiovascular disease. Our objective was to determine the prevalence of diastolic dysfunction in HIV-infected patients without cardiovascular symptoms. We enrolled 91 subjects in a cross-sectional study of HIV-infected patients without cardiovascular symptoms between September 2004 and August 2005, to assess whether demographics, HIV-related factors, cardiac risk factors, and ART were associated with diastolic dysfunction. All subjects underwent two-dimensional transthoracic echocardiography with tissue Doppler imaging. Subjects were predominately male with a median age of 38 (interquartile range [IQR]: 33, 42) years and median ART duration 6.15 (IQR: 2.1, 8.4) years. Subjects had low Framingham risk scores. Diastolic dysfunction was observed in 34 patients (37%; 95% confidence interval [CI] 27.4, 48.1). Cardiac risk factors or poor prognostic indicators of AIDS progression were uncommon with no difference between subjects with or without diastolic dysfunction. A nonstatistically significant trend in increased rate of diastolic dysfunction was observed in patients receiving protease inhibitors 1 year or more, 44% versus 28%, respectively (univariate odds ratio 2.02, 95% CI 0.83 to 4.90). This was not observed with prolonged use of either non-nucleoside or nucleoside reverse transcriptase inhibitors. A high prevalence of diastolic dysfunction (37%) in a cohort of HIV-infected patients on ART at low risk for AIDS and cardiovascular disease was demonstrated. PMID- 19281431 TI - AIDS orphanages in China: reality and challenges. AB - With the increasing number of AIDS orphans in China, the government has been building AIDS orphanages since 2004 to accommodate some of those children who have lost both parents to AIDS. However, no data are available regarding the quality of this model of institutional care of AIDS orphans in China. This study, based on qualitative data from children and workers in AIDS orphanages, examines the daily lives, needs, and feelings of orphans and explores the advantages and disadvantages of institutionalized care of AIDS orphans in China. The current study was conducted in 2006-2007 in two rural counties of central China. Data in the current study included individual in-depth interviews with 23 children who lost both of their parents to HIV/AIDS (ages 8 to 17 years) living in AIDS orphanages and 5 AIDS orphanage workers. Findings in this study reveal that children living in orphanages mostly felt that the living conditions were better than the families they lived with after the death of their parents. However, according to the children and orphanage workers, the institutional care has some disadvantages, such as administrative restraints, limited psychological guidance, stigma, lack of education on AIDS, and financial burdens of the operation. Implications for intervention programs include continuing support from the government and nongovernmental organizations, improvements in administrative styles, and the need of incorporating psychological support within the institutions. PMID- 19281432 TI - Preloading potential of retroviral vectors is packaging cell clone dependent and centrifugation onto CH-296 ensures highest transduction efficiency. AB - Retroviral vector-mediated gene transfer has been used successfully in clinical gene therapy. Cells of the hematopoietic lineages, however, remain difficult to transduce, although precoating of culture vessels with the fibronectin fragment CH-296 may improve transduction efficiency. Alternatively, low-speed centrifugation of vector-containing supernatant onto culture vessels may improve transduction efficiency in the absence of CH-296 preloading. Using the NIH/3T3 derived Moloney murine leukemia virus-based packaging cell lines PG13, PA317, and PT67, we here show that preloading by low-speed centrifugation improves transduction efficiency in a packaging cell subclone-dependent manner. Preloading by centrifugation, however, cannot generally replace CH-296 and we obtained the overall highest transduction levels when combining centrifugation and CH-296 precoating. We found, moreover, that the factor responsible for high susceptibility to preloading in our PG13-derived vector supernatant was transferable to a PA317-derived vector supernatant with low susceptibility to preloading. Furthermore, our PA317, PG13, and PT67 subclones shed into their supernatants variable amounts of fibronectin. This soluble fibronectin formed aggregates of various sizes and generated complexes with vector particles. The fibronectin-vector complexes readily sedimented onto culture vessels and copurified after fibronectin-specific affinity purification of vector-containing supernatants. Finally, vector supernatant from 293T cells, which barely produce fibronectin, was not susceptible to preloading. The susceptibility to preloading by centrifugation thus appears to be dependent both on the specific packaging cell line and on the association of vector particles and packaging cell-produced fibronectin. Rigorous screening of individual vector-containing supernatants is therefore required to identify optimal transduction conditions for retroviral gene transfer. PMID- 19281433 TI - West Nile virus activity in mosquitoes and domestic animals in Chiapas, Mexico. AB - Prior to 2006, West Nile virus (WNV) had not been definitively detected in Chiapas, the southernmost state of Mexico, although it circulates elsewhere in Mexico and Central America. We collected over 30,000 mosquitoes and blood-sampled 351 domestic animals in Chiapas in search for evidence of current or recent transmission of WNV. Two mosquito pools tested positive for WNV RNA and 17 domestic animals tested positive for specific WNV-neutralizing antibodies, including young animals (<1 year old) in four of five sampled locations. The two WNV-positive mosquito pools were collected on the Pacific coastal plain of Chiapas in June, 2006, and included a pool of Culex nigripalpus, a suspected vector of WNV, and a pool of Cx. interrogator. The sequence of a 537-nucleotide portion of a cDNA amplicon derived from the WNV NS5 gene from the Cx. interrogator pool contained a single silent nucleotide substitution when compared to WNV strain NY99. PMID- 19281434 TI - Genetic variation associated with mammalian feeding in Culex pipiens from a West Nile virus epidemic region in Chicago, Illinois. AB - Mosquitoes of the Culex pipiens complex are important vectors of West Nile virus in the United States. We examined the genetic variations of Cx. pipiens mosquitoes from Chicago, Illinois that were determined to be principally ornithophilic but exhibited a relatively higher inclination for mammalian hosts including humans. Microsatellite analysis of 10 polymorphic markers was performed on 346 engorged Cx. pipiens specimens with identified avian or mammalian blood meals. Our results indicated that there were no significant differences in allelic richness, the pattern of conformity to Hardy-Weinberg equilibrium, and linkage disequilibrium, nor was there overall genetic differentiation between specimens with avian- and mammalian-derived blood meals. However, Cx. pipiens form pipiens with mammalian- (including human-) derived blood meals had significantly higher ancestry (p < 0.001) and proportion of hybrids (p < 0.01) from the Cx. pipiens form molestus (population from New York City) than did those with avian-derived blood meals. By contrast, there were no significant differences in the ancestry (p > 0.05) and the proportion of hybrids (p > 0.05) from Cx. quinquefasciatus (population from Harris Country, Texas). No temporal genetic variation was detected in accordance with the observation that there was no shift in blood feeding from birds to mammals. The results of this study in conjunction with regional host-feeding behavior suggest that the probability of genetic ancestry from Cx. pipiens f. molestus may predispose mosquitoes to feed more readily on mammals; however, the genetic mechanisms are unknown. PMID- 19281435 TI - Comparative analysis of Trypanosoma rangeli histone H2A gene intergenic region with distinct intraspecific lineage markers. AB - This study shows the characterization of the histone H2A intergenic region sequences (H2A IR) from Trypanosoma rangeli KP1(+) and KP1(-) strains isolated from distinct hosts and geographic regions. Also, a comparative unweighted pair group method using arithmetic averages (UPMGA) analysis with polymerase chain reaction profiles of the 24Salpha rDNA and the miniexon genes was performed. Detailed H2A IR sequence analysis revealed a discrete size polymorphism among T. rangeli strains and the presence of single-nucleotide polymorphisms and minisatellite repeats, exclusively allowing an interspecific differentiation from T. cruzi strains representing the main parasite lineages. Differently from the H2A IR, UPMGA analysis of the 24Salpha rDNA and the miniexon genes profiles clearly branched T. rangeli strains into KP1(-) and KP1(+) lineages, clustering separately the Brazilian and Colombian KP1(-) strains. The evolutionary implications of these findings are discussed. PMID- 19281436 TI - Tracheostomy teams - filling a void. PMID- 19281437 TI - Why publish statistical analysis plans? PMID- 19281438 TI - The relationship between blood glucose level and QTc duration in the critically ill. AB - OBJECTIVE: To determine whether hyperglycaemia is associated with prolongation of the corrected QT (QTc) interval on the electrocardiogram (ECG) in critically ill patients. DESIGN: Single-centre, prospective observational study. PARTICIPANTS AND SETTING: 197 consecutive patients admitted to the adult intensive care unit of a 570-bed teaching hospital over 10 weeks from November 2004. MAIN OUTCOME MEASURES: Correlation between QT interval (on standard 12-lead ECG taken on ICU admission, corrected with Bazett's formula) and serum glucose level (BGL) in blood collected at time of ECG; comparison of variables, including BGL, by QTc category (< or = 0.44 s or > 0.44 s); explained variance (R(2)) of QTc, determined by multivariate regression analysis. RESULTS: Mean patient age was 53.4 years. A moderate, positive correlation was found between QTc and BGL (Pearson's correlation coefficient, r = 0.277, P < 0.001). A standard multivariate regression model explained 32.9% (R(2)) of QTc variance, and revealed four significant, independent predictors of QTc duration: heart rate (explaining 11.4% of QTc variance), use of inotropes (10.1%), BGL (7.3%) and serum magnesium level (4.6%). In the cohort with QTc > 0.44 s, BGL was significantly higher, as were the need for inotropes, APACHE II scores and mortality. QTc was significantly longer in patients with BGL > 8 mmol/L than in those with lower BGL (0.471 v 0.442 s, P < 0.001). The only independent predictors of mortality were APACHE II score and mean arterial pressure. CONCLUSIONS: There was a moderate, significant correlation between QTc and BGL. Patients with a QTc > 0.44 s had higher BGL, APACHE II score and mortality. BGL was an independent predictor of QTc duration, but neither BGL nor QTc were independent predictors of mortality in this study. PMID- 19281439 TI - Outcomes of patients with spinal cord injury before and after introduction of an interdisciplinary tracheostomy team. AB - OBJECTIVES: To assess outcomes in patients with spinal cord injury (SCI) and a tracheostomy tube (TT), before and after the introduction of a tracheostomy review and management service (TRAMS) for ward-based patients. DESIGN: Matched pairs design with two cohorts, before and after the intervention. SETTING: 900 bed tertiary hospital in Melbourne, Victoria. PARTICIPANTS: SCI patients with a TT that was removed: 34 patients in the post-TRAMS period (September 2003 to September 2006) were matched to 34 from the pre-TRAMS period (September 1999 to December 2001). INTERVENTION: TRAMS was introduced as a consultative team of specialist physicians, clinical nurse consultants, physiotherapists and speech pathologists. The team coordinated tracheostomy care, conducted twice-weekly rounds, and provided policy, education, and support. MAIN OUTCOME MEASURES: Comparison of length of stay (LOS), duration of cannulation (DOC), improved communication through use of a one-way valve, number of adverse events and related costs. RESULTS: Median patient LOS decreased from 60 days (interquartile range [IQR], 38-106) to 41.5 days (IQR, 29- 62) (P = 0.03). The pre-TRAMS median DOC decreased from 22.5 days (IQR, 17-58) to 16.5 days (IQR, 12-25) (P = 0.08). Speaking-valve use increased from 35% (12/34) to 82% (28/34) (P < 0.01). Median time to a valve trial decreased from 22 days (IQR, 13-44) to 6 days (IQR, 4-10) after TT insertion (P < 0.01). There were two tracheostomy-related medical emergency calls pre-TRAMS and none post-TRAMS. There were no tracheostomy-related deaths in either group. The annual cost savings from implementing TRAMS were about eight times greater than the cost of service provision. CONCLUSION: Implementing a tracheostomy review and management service improved outcomes for SCI patients: they left acute care sooner, spoke sooner, and the TT was removed earlier, with associated cost savings. PMID- 19281440 TI - Operator anaesthesiology training and complications after endotracheal intubation in the intensive care unit: a 3-year, prospective, observational study. AB - OBJECTIVE: To determine whether operators with less than 3 months' formal anaesthesiology training have higher rates of complications when performing endotracheal intubation (ETI) in the intensive care unit than operators with longer formal anaesthesiology training. DESIGN AND SETTING: Prospective, single centre, observational study of consecutive ETIs performed in a general, urban, tertiary ICU between May 2005 and May 2008. Data were collected by self-reported, written questionnaire. PARTICIPANTS: The two pre-defined study cohorts were ETIs performed where the initial operator had less than 3 months' formal training in anaesthesiology, and those where the initial operator had 3 months' or longer training. MAIN OUTCOME MEASURES: The primary outcome measure was the number of ETIs where one or more pre-defined complications occurred as a result of the ETI. Secondary outcome measures were the number of ETIs where one or more respiratory, cardiovascular or trauma complications occurred as a result of the ETI, and the number where the airway was deemed difficult by the operator. RESULTS: Data were collected on 276 ETIs. There were no significant differences in primary or secondary outcome measures between the two main study groups. Operators with less than 3 months' formal training in anaesthesiology had a higher level of medical supervision or assistance (75% v 29%, P<0.001), more favourable patient pre intubation oxygen saturation on pulse oximetry (SpO(2)) (76% v 65% had SpO(2)>89%, P=0.05), and easier resultant grade of intubation (70% v 56% of intubations were Grade I, P= 0.04), but required more operators (19% v 3% required two operators, P<0.001), and more attempts before ETI was successful (62% v 82% of intubations were successful on first attempt, P<0.001). CONCLUSION: ETIs performed in the ICU where the initial operator has less than 3 months' formal training in anaesthesiology appear not to be associated with more complications. However, this may be attributable to less experienced operators having more assistance and supervision, and to patient selection. PMID- 19281441 TI - Effect of implementation of a weekly multidisciplinary team meeting in a general intensive care unit. AB - OBJECTIVE: To determine whether the introduction of a weekly multidisciplinary team meeting (MDTM) to a general intensive care unit improved selected clinical indicators of patient outcome, and staff satisfaction with patient care. DESIGN: A single-centre, observational, before-and-after study. SETTING: A 14-bed general ICU in an urban, tertiary teaching hospital. STUDY POPULATION: All patients admitted to the ICU during June-December 2006 (before the intervention) and June- December 2007 (after the intervention), and staff employed in the ICU in December 2006 and December 2007. INTERVENTION: Introduction of a weekly MDTM to the ICU. MAIN OUTCOME MEASURES: The primary outcome was the number of patients who stayed in the ICU longer than 5 days. Secondary outcomes included nurses' scores for satisfaction with patient care on a questionnaire; ICU and hospital mortality; duration of mechanical ventilation; readmissions to the ICU within 72 hours of discharge; and after-hours discharges. RESULTS: There were 376 ICU admissions in the "before" period and 432 in the "after" period. Baseline characteristics of the two groups were similar except for a lower proportion of patients admitted directly to the ICU from the operating theatres in the after period (34.2% v 45.2%, P = 0.002). There were no significant differences in any of the primary or secondary outcomes, with the exception of one questionnaire score: a fall in the score nursing staff gave for value of all meetings held in the ICU following the introduction of the MDTM (from 6.6 to 3.9 on a scale of 0-10, P = 0.001). CONCLUSION: The introduction of a weekly MDTM to a general ICU did not improve selected clinical indicators of patient outcome or staff satisfaction with patient care. PMID- 19281442 TI - Femoral-radial arterial pressure gradients in critically ill patients. AB - OBJECTIVE: To investigate the presence and determinants of femoral-radial gradients in mean arterial pressure (MAP) in a critically ill population. DESIGN: Prospective observational study. SETTING AND PARTICIPANTS: Critically ill patients who were undergoing simultaneous monitoring of arterial pressure by radial arterial catheterisation and transpulmonary thermodilution (via femoral arterial access) in a Level 3, mixed medical-surgical intensive care unit, December 2007 to May 2008. MAIN OUTCOME MEASURES: Level of agreement between simultaneous measurements of MAP via the femoral and radial arteries, determined by Bland-Altman analysis; haemodynamic and demographic factors associated with a MAP gradient, assessed by multiple linear regression. RESULTS: 131 observations were made in 24 patients. Mean age of patients was 56 (SD, 18) years, and mean APACHE II score was 27 (SD, 8). Overall mean bias between radial and femoral MAP measurements was 4.27 mmHg (limits of agreement, -3.41 to 11.94 mmHg). Fifteen patients (62%) had maximum MAP gradients > 5mmHg, and seven of these (29% of the total) had maximum gradients > 10 mmHg. The largest discrepancy in MAP was 18 mmHg in a patient with septic shock resistant to high-dose catecholamine infusion. Regression analysis failed to identify any statistically significant associations between patient factors and MAP gradient. CONCLUSION: A systematic difference in MAP measured at the radial and femoral sites was demonstrated. In some critically ill patients, the femoral artery may be the preferred site for systemic arterial pressure monitoring. PMID- 19281443 TI - Post-hypoxic myoclonic status: the prognosis is not always hopeless. AB - A 67-year-old woman abruptly developed acute pulmonary oedema, severe bradycardia and then cardiac arrest while in hospital 6 days after an elective hernia repair. She was resuscitated, intubated and transferred to the intensive care unit. Within 24 hours, she began to display repetitive, generalised myoclonic jerks that failed to respond to therapy with conventional anticonvulsants; an electroencephalogram confirmed myoclonic status. After administration of levetiracetam was begun on Day 3, myoclonic jerks reduced, and there was gradual clinical improvement. By Day 6 after the arrest, the patient was alert and oriented (Glasgow Coma Score, 15/15). Although she died on Day 11 after massive haemoptysis and cardiac arrest, this patient demonstrates the possibility of reasonable neurological recovery despite early onset of myoclonic status. PMID- 19281444 TI - Survival after a massive overdose of arsenic trioxide. AB - Arsenic poisoning remains a therapeutic challenge, and outcomes are often poor. An 18-year-old man deliberately ingested termiticide containing a massive dose of arsenic trioxide. Arsenic concentration was 6.3 micromol/L in serum on ICU Day 1, and 253 micromol/L in the first 24-hour urine sample, with a urinary arsenic/creatinine ratio of 84 200 micromol/mol. He was treated with the chelating agent meso-2,3-dimercaptosuccinic acid (DMSA) (replaced by dimercaprol on Days 2-5) and required intensive support for multisystem organ failure, but recovered slowly. Nine weeks after the ingestion the only ongoing clinical issue was persistent but slowly improving peripheral neuropathy. PMID- 19281445 TI - The NICE-SUGAR (Normoglycaemia in Intensive Care Evaluation and Survival Using Glucose Algorithm Regulation) Study: statistical analysis plan. AB - BACKGROUND: The Normoglycaemia in Intensive Care Evaluation and Survival Using Glucose Algorithm Regulation (NICE-SUGAR) Study is the largest study to date of glycaemic control in critically ill patients. OBJECTIVE: To describe in detail and make public the study's pre-determined statistical analysis plan, which was finalised while data collection was still ongoing, and to which the investigators will adhere in analysing the data from the trial. METHODS: The data collected by researchers as part of the trial protocol were reviewed and formally assessed. Information relevant to baseline characteristics was selected and, for each item, statistically relevant descriptive elements were described. Information relevant to the process of care and delivery of prescribed trial therapy was similarly classified and, for each item, appropriate descriptive statistical analysis was planned with appropriate comparison between groups. Finally, trial outcomes were classified as primary, secondary or tertiary, and an appropriate statistical comparison between groups was planned and described. RESULTS: A standard analysis plan was developed for the results of the NICE-SUGAR Study. This plan allows a comprehensive description of baseline characteristics, features of the process of care, and trial treatment delivery, along with pre-determined statistical assessment of relevant outcome measures in a way that is transparent, available to the public, verifiable and pre-determined before completion of data collection. CONCLUSION: We have developed a pre-determined statistical analysis plan for the NICE-SUGAR Study. This plan will be followed to avoid analysis bias arising from prior knowledge of the study findings. PMID- 19281446 TI - The RENAL (Randomised Evaluation of Normal vs. Augmented Level of Replacement Therapy) study: statistical analysis plan. AB - BACKGROUND: The Randomised Evaluation of Normal vs. Augmented Level of Replacement Therapy (RENAL) study is the largest interventional trial ever conducted in patients with acute renal failure. OBJECTIVE: To develop and report a pre-determined statistical analysis plan which the investigators will adhere to in analysing the data from the trial. METHODS: The data collected by the researchers as part of the trial protocol was reviewed and formally assessed. Information relevant to baseline characteristics was selected and, for each item, statistically relevant descriptive elements were described. Information relevant to the process of care and delivery of prescribed trial therapy was similarly classified and, for each item, appropriate descriptive statistical analysis was planned with appropriate comparison between groups. Finally, trial outcomes were selected, and an appropriate statistical comparison between groups was planned and described. RESULTS: A standard analysis plan for the RENAL trial results was developed, which allows a comprehensive description of baseline characteristics, features of the process of care and trial treatment delivery, and pre-determined statistical assessment of relevant outcome measures in a way that is transparent, available to the public, verifiable and predetermined before the actual analysis of data. CONCLUSION: We have developed a pre-determined statistical analysis plan for the RENAL trial. This plan will be adhered to in order to avoid introducing any analysis bias associated with prior knowledge of study findings. PMID- 19281447 TI - Albumin is a blood product too - is it safe for all patients? AB - Albumin has been used for volume resuscitation and supplementation in critically ill patients for over 50 years. While regarded as a "gold standard" colloid solution, albumin is associated with substantial cost, and questions have been raised about its safety and efficacy. A large-scale randomised controlled trial (the Saline vs. Albumin Fluid Evaluation [SAFE] study) demonstrated that albumin and saline were clinically equivalent treatments for intravascular volume resuscitation in a heterogenous population of critically ill patients. However, in patients with traumatic brain injury, albumin was associated with a significantly higher mortality and cannot be recommended for acute resuscitation of such patients. A potential beneficial role of albumin in patients with severe sepsis, particularly malaria, requires further study. Extrapolation of the results of the SAFE study to other, synthetic, colloid solutions requires caution, and a randomised controlled trial comparing albumin, starch and crystalloids in patients with severe sepsis is warranted. The safety of synthetic colloids in patients with traumatic brain injury should not be assumed. Although hypoalbuminaemia is associated with increased mortality, use of albumin for volume resuscitation of critically ill patients with a serum albumin concentration < or =25 g/L is not associated with reductions in mortality, duration of ICU stay or mechanical ventilation, or in use of renal replacement therapy. Similarly, there is no substantive evidence to justify the use of hyperoncotic albumin solutions for resuscitation or supplementation in critically ill patients. Albumin is a safe and effective resuscitation solution in critically ill patients without traumatic brain injury. However, the acquisition costs of albumin and synthetic colloids are more than those of crystalloids, and, as yet, colloids have not been proven to confer substantive benefits over crystalloids such as saline. PMID- 19281448 TI - To transfuse, or not to transfuse: that is the question. AB - Transfusion practice, like medical practice in its entirety, has evolved rapidly over the past few decades. Recent research on clinical outcomes has examined the impact of blood transfusion on critically ill patients, patients with trauma, those undergoing cardiac surgery, those experiencing acute coronary syndromes, oncology patients and others. Evidence is mounting of adverse outcomes associated with blood transfusion in a wide variety of clinical contexts. Here, we highlight the deficit in the current literature guiding transfusion practice, and call for an Australasian study to fill this deficit. PMID- 19281449 TI - 19th century pioneering of intensive therapy in North America. Part 3: the Fell O'Dwyer apparatus and William P Northrup. AB - Two previous articles in this series have described the reintroduction of forced respiration for ventilatory difficulties, particularly in opiate poisoning (by George Fell), and successful use of intralaryngeal tubes designed for treating airway obstruction in diphtheritic acute laryngitis (by Joseph O'Dwyer). In 1891, O'Dwyer extended the applications of Fell's system, introducing a longer orolaryngeal tube, replacing Fell's methods of inflating the lungs, which had been with a somewhat unsatisfactory facemask or through a tracheotomy tube. The combined system became known as the "Fell-O'Dwyer apparatus". Use of the apparatus widened, to include treating apnoea from intracranial disasters (by William P Northrup, especially) and, on the initiative of Rudolph Matas, in delivering anaesthesia and maintaining lung inflation to enable intrathoracic surgery (by FW Parham). Although the apparatus was used beyond New York (eg, in New Orleans by J D Bloom, especially for neonatal apnoea), it is difficult to find other than nonspecific references. Matas and Bloom improved O'Dwyer's original system, but after the clinical success of Charles Elsberg's continuous insufflation anaesthesia for thoracic surgery, 1909, American anaesthetists came to prefer that. PMID- 19281450 TI - Production and metabolic engineering of terpenoid indole alkaloids in cell cultures of the medicinal plant Catharanthus roseus (L.) G. Don (Madagascar periwinkle). AB - The Madagascar periwinkle [Catharanthus roseus (L.) G. Don] is a plant species known for its production of TIAs (terpenoid indole alkaloids), many of which are pharmaceutically important. Ajmalicine and serpentine are prescribed for the treatment of hypertension, whereas the bisindoles vinblastine, vincristine and 3',4'-anhydrovinblastine are used for their antineoplastic activity in the treatment of many cancers. However, TIAs are produced in small yields in C. roseus, which make them expensive. Cell and metabolic engineering has focused on increasing flux through the TIA pathway by various means, including optimization of medium composition, elicitation, construction of noval culture systems and introduction of genes encoding specific metabolic enzymes into the C. roseus genome. The present review will attempt to present the state-of-the-art of research in this area and provide an update on the cell and metabolic engineering of TIAs in C. roseus. We hope that this will contribute to a better understanding of the ways in which TIA production can be achieved in different C. roseus culture systems. PMID- 19281451 TI - Sympatho-vagal responses in patients with sleep and typical vasovagal syncope. AB - Sleep syncope is a recently described form of vasovagal syncope that interrupts sleep. The pathophysiology of this condition is uncertain but a 'central' non baroreflex-mediated trigger has been suggested. In the present study, we tested the hypothesis that patients with sleep syncope have abnormal sympatho-vagal responses to non-baroreflex, but normal responses to baroreflex stimuli. We collected historical data from SS patients (patients with vasovagal syncope with sleep syncope; n=16) and NSS patients (patients with vasovagal syncope without sleep syncope; n=35), including demography, and triggers and symptoms during syncope. MBP (mean blood pressure), HR (heart rate) and MSNA (muscle sympathetic nerve activity) in SS patients were compared with NSS patients and matched controls (n=16) during HG (handgrip), CPTs (cold pressor tests), HUT (head-up tilting) and tilt-induced pre-syncope. Patients and controls were of similar age and gender distribution [SS patients, age 46.0+/-4 years (69% female); NSS patients, 47.3+/-4 years (63% female); controls, 43.7+/-5 years (69% female)]. Compared with NSS patients, SS patients reported more fainting episodes: (i) triggered by phobias (75 compared with 37%; P=0.001); (ii) while in the horizontal position (44 compared with 6%; P=0.001); and (iii) associated with abdominal symptoms (69 compared with 9%; P=0.001). Compared with controls, the MBP response to HG was attenuated in SS patients (P=0.016), and MSNA (burst frequency and incidence) responses to CPT were attenuated in both syncope groups (SS, P=0.011 and 0.003 respectively; NSS, P=0.021 and 0.049 respectively). MSNA responses to HUT did not differ. For both non-baroreflex and baroreflex responses, there were no differences in any of the MSNA indices between the syncope groups. Patients with vasovagal syncope, with or without sleep syncope, have very similar sympatho-vagal responses to both non-baroreflex and baroreflex stimuli. This is consistent with sleep syncope being a subform of vasovagal syncope. Attenuation of sympathetic responses to non-baroreflex pathways may be important in the mechanism of vasovagal syncope. PMID- 19281452 TI - Proton transfer in the mechanism of polyadenylate polymerase. AB - PAP (polyadenylate polymerase) is the template-independent RNA polymerase responsible for synthesis of the 3' poly(A) tails of mRNA. To investigate the role of proton transfer in the catalytic mechanism of PAP, the pH dependence of the steady-state kinetic parameters of yeast PAP were determined for the forward (adenyl transfer) and reverse (pyrophosphorolysis) reactions. The results indicate that productive formation of an enzyme-RNA-MgATP complex is pH independent over a broad pH range, but that formation of an active enzyme-RNA MgPPi complex is strongly pH dependent, consistent with the production of a proton on the enzyme in the forward reaction. The pH dependence of the maximum velocity of the forward reaction suggests two protonic species are involved in enzyme catalysis. Optimal enzyme activity requires one species to be protonated and the other deprotonated. The deuterium solvent isotope effect on Vmax is also consistent with proton transfer involved in catalysis of a rate-determining step. Finally, pKa calculations of PAP were performed by the MCCE (multiconformational continuum electrostatic) method. Together, the data support that the protonation of residues Lys215 and Tyr224 exhibit co-operativity that is important for MgATP2 and MgPPi2- binding/dissociation, and suggest these residues function in electrostatic, but not in general acid, catalysis. PMID- 19281453 TI - Cross-talk between the VEGF-A and HGF signalling pathways in endothelial cells. AB - BACKGROUND INFORMATION: Endothelial cells play a major role in angiogenesis, the process by which new blood vessels arise from a pre-existing vascular bed. VEGF-A (vascular endothelial growth factor-A) is a key regulator of angiogenesis during both development and in adults. HGF (hepatocyte growth factor) is a pleiotropic cytokine that may promote VEGF-A-driven angiogenesis, although the signalling mechanisms underlying this co-operation are not completely understood. RESULTS: We analysed the effects of the combination of VEGF-A and HGF on the activation of VEGFR-2 (VEGF receptor-2) and c-met receptors, and on the stimulation of downstream signalling pathways in endothelial cells. We found that VEGFR-2 and c met do not physically associate and do not transphosphorylate each other, suggesting that co-operation involves signalling events more distal from receptor activation. We demonstrate that the VEGF isoform VEGF-A(165) and HGF stimulate a similar set of MAPKs (mitogen-activated protein kinases), although the kinetics and strengths of the activation differ depending on the growth factor and pathway. An enhanced activation of the signalling was observed when endothelial cells were stimulated by the combination of VEGF-A(165) and HGF. Moreover, the combination of VEGF-A and HGF results in a statistically significant synergistic activation of ERK1/2 (extracellular-signal-regulated kinase 1/2) and p38 kinases. We demonstrated that VEGF-A(165) and HGF activate FAK (focal adhesion kinase) with different kinetics and stimulate the recruitment of phosphorylated FAK to different subsets of focal adhesions. VEGF-A(165) and HGF regulate distinct morphogenic aspects of the cytoskeletal remodelling that are associated with the preferential activation of Rho or Rac respectively, and induce structurally distinct vascular-like patterns in vitro in a Rho- or Rac-dependent manner. CONCLUSIONS: Under angiogenic conditions, combining VEGF-A with HGF can promote neovascularization by enhancing intracellular signalling and allowing more finely regulated control of the signalling molecules involved in the regulation of the cytoskeleton and cellular migration and morphogenesis. PMID- 19281454 TI - Molecular detection and identification of Rickettsia endosymbiont in different biotypes of Bemisia tabaci. PMID- 19281455 TI - Potential vectors and hosts of rickettsia spp: epidemiological studies in the Vale do Paraiba, state of Rio de Janeiro/Brazil. PMID- 19281456 TI - Uveitis in the course of Q-fever. PMID- 19281457 TI - DNA microarray-chip based diagnosis of Q-fever (Coxiella burnetii). PMID- 19281458 TI - Serological and molecular characterization of AdaA: a potential marker of Q-fever abortion in goats? PMID- 19281459 TI - Role of specific antibodies in Coxiella burnetii infection of macrophages. PMID- 19281460 TI - Carriage of Rickettsia spp., Coxiella burnetii and Anaplasma spp. by endemic and migratory wild birds and their ectoparasites in Cyprus. PMID- 19281461 TI - A systematic approach to evaluate humoral and cellular immune responses to Coxiella burnetii immunoreactive antigens. PMID- 19281462 TI - Comparing two types of botulinum-A toxin detrusor injections in patients with severe neurogenic detrusor overactivity: a case-control study. AB - OBJECTIVE: To compare the efficacy of two types of botulinum toxin type A (BTX-A; Dysport, Ipsen Ltd, Slough, UK) or Botox (Allergan Inc., Irvine, CA, USA) and examine the possible dose-effect relation for Dysport in those patients, as multifocal detrusor injections with BTX-A are effective for severe neurogenic detrusor overactivity in adults. PATIENTS AND METHODS: This was an open-label, observational case-control study comparing Dysport and Botox, and the dose-effect relation for Dysport. The patients included were treated with either Dysport (cases; 500, 750, or 1000 IU), or with Botox (controls: 300 IU, and exceptionally 200 or 400 IU). The continence volume (primary), mean and maximum catheterized volume, and antimuscarinic use were assessed, and cystometric variables measured were overactivity volume (primary), detrusor compliance, and cystometric capacity. All variables were assessed at baseline, 3 and 9 months after treatment, and results analysed using analysis of variance (paired) t-tests, chi square tests and regression analysis. RESULTS: There were 28 cases and 28 matched controls; their demographic characteristics, bladder management, and baseline values were comparable between the groups. At 3 months there was a significant improvement in cystometric variables and continence volume in both groups, but in mean catheterized volume and reduced use of antimuscarinics in cases only. At 9 months there was no significant improvement over baseline except for the continence volume in the cases. There were no significant differences in results between the groups except for the continence volume at 3 months (459 mL after Dysport and 396 mL after Botox; P=0.015). There was no dose-related response for Dysport at 3.8 months of follow-up. The interval between injections (9.5, 14.5 and 16.1 months for Dysport 500, 750, and 1000 IU; 10.1 months for Botox) was not significant. There were nine non-responders in the Dysport group and seven in the Botox group; the patient characteristics and baseline data were comparable to those of the responders. There was transient hypoasthenia in one of the responders (750 IU Dysport). CONCLUSIONS: A single treatment session with either Dysport or Botox in a setting combined with antimuscarinics might improve the patient's condition for up to a year. There was no clear dose-related effect for Dysport in adults. PMID- 19281463 TI - Long-term effects of intermittent androgen suppression on testosterone recovery and bone mineral density: results of a 33-month observational study. AB - OBJECTIVE: To investigate changes in bone mineral density (BMD) and osteoporosis, over 3 years of intermittent androgen-suppression therapy (IAST). PATIENTS AND METHODS: This was a Phase II individual cohort study of 72 patients with prostate cancer without metastatic bone disease, enrolled between 1999 and 2002. Patients had 9 months flutamide (250 mg, three times daily) and leuprolide (22.5 mg, 3 monthly depot) after which, patients ceased therapy providing that their PSA levels were <4 ng/mL. AST re-commenced when the PSA level exceeded the pretreatment level or was >20 ng/mL. BMD for hip and spine was the primary endpoint; assessed at baseline; completion of initial treatment period; and at 1 and 2 years after initial treatment (POST period). RESULTS: Osteoporosis increased from 7% at baseline to 10% at 3 years. The BMD declined after 9 months treatment, at -1.9% and -3.3% at hip and spine, respectively (P < 0.001). Subsequent BMD decline in the POST period was attenuated; at 1 years and 2 years later, hip -0.6% (not significant), and -0.8% (P < 0.014), and spine +1.0% and +0.2% (not significant). The BMD change in those remaining 'off' therapy for 2 years (n = 20) was strongly associated with the level of testosterone recovery; a peak testosterone level of <5 nmol/L associated with a greater then normal physiological loss. Testosterone recovery was less likely in older men. CONCLUSION: The attenuation of spine and hip BMD decline after 3-year IAST compared with those reported for continuous AST appears to be due to testosterone driven BMD recovery in the POST period. Failure of testosterone recovery was associated with worse final BMD. By reducing the potential risk for adverse bone complications, intermittent therapy may become an important consideration when the therapeutic ratio is narrow. PMID- 19281464 TI - Description of magnetic resonance imaging-derived enhancement variables in pathologically confirmed prostate cancer and normal peripheral zone regions. AB - OBJECTIVE: To assess the use of a semiquantitative analysis of dynamic contrast enhanced (DCE) magnetic resonance imaging (MRI) to produce indices for enhancement curves that might enable differentiation between malignant prostatic lesions and normal peripheral zone (PZ). PATIENTS AND METHODS: Fifty-two patients scheduled for radical prostatectomy underwent DCE-MRI before surgery using a 3 T scanner. The DCE images were used to generate variables from changes in signal intensity for pathologically confirmed malignant areas and the normal PZ, using whole-mounted pathology specimens as a reference to delineate regions of interest (ROI). These variables included maximum enhancement index (MaxEI), time to MaxEI at 30 s, the initial and final slopes of signal intensity change, and the area under curve. A threshold value for each DCE variable was identified, and the sensitivity and specificity were obtained. RESULTS: Malignant lesions had a 56% higher MaxEI than normal PZ and took half the time to reach MaxEI (P<0.001). Hence, at 30 s, cancer lesions have double the mean (sd) EI than normal PZ, of 2.22 (1.04) vs 1.04 (0.51), respectively. Tumours showed significant washout of contrast medium, which was reflected in the final slope of the curve being negative, as opposed to positive for normal PZ. The combined data of DCE variables, using a logistic regression test, gave a mean (95% confidence interval) sensitivity and specificity of 89 (81-96)% and 90 (83-97)%, respectively. CONCLUSION: This technique provides good discrimination of malignant lesions that might enable accurate localisation of the lesion. It is a simple, semiquantitive, noninvasive method that reflects the unusual vascular characteristics of newly formed microvessels and the changes in the interstitium that occur in prostate cancer. PMID- 19281465 TI - Prospective evaluation of hybrid 18F-fluorodeoxyglucose positron emission tomography/computed tomography in staging clinically node-negative patients with penile carcinoma. AB - OBJECTIVE: To prospectively evaluate the performance of 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) to detect occult metastasis in patients with clinically node-negative (cN0) penile carcinoma, as there is little information on the use of (18)F-FDG-PET/CT in penile carcinoma. PATIENTS AND METHODS: In 24 patients, scheduled to undergo dynamic sentinel-node biopsy, hybrid PET/CT was used before surgery to assess the nodal status of the cN0-groins. Six of the 24 patients were unilaterally cN0. Thus, 42 cN0-groins were evaluated for occult metastasis using PET/CT. All scans were assessed by two experienced nuclear physicians. The histopathological tumour status of the removed sentinel node was used as the standard of care to evaluate the PET/CT-results. RESULTS: Histopathology was tumour-positive in five of the 42 (12%) evaluated cN0-groins, two of which contained only micrometastases (<2 mm). One of the five tumour-positive cN0-groins was correctly predicted on the PET/CT images. All false-negative PET/CT scans contained metastasis of or =70% of participants except for terminal dribble in men and split stream in women. CONCLUSION: In this large population study of three countries, LUTS are highly prevalent among men and women aged >40 years. In general, LUTS experienced 'often' or more are bothersome to most people. PMID- 19281468 TI - A prospective, non-randomized trial comparing robot-assisted laparoscopic and retropubic radical prostatectomy in one European institution. AB - OBJECTIVE: To compare the functional results of two contemporary series of patients with clinically localized prostate cancer treated by robot-assisted laparoscopic prostatectomy (RALP) or retropubic radical prostatectomy (RRP). PATIENTS AND METHODS: This was a non-randomized prospective comparative study of all patients undergoing RALP or RRP for clinically localized prostate cancer at our institution from February 2006 to April 2007. RESULTS: We enrolled 105 patients in the RRP and 103 in the RALP group; the two groups were comparable for all clinical and pathological variables, except median age. For RRP and RALP the respective median operative duration was 135 and 185 min (P < 0.001), the intraoperative blood loss 500 and 300 mL (P < 0.001) and postoperative transfusion rates 14% and 1.9% (P < 0.01). There were complications in 9.7% and 10.4% of the patients (P = 0.854) after RRP and RALP, respectively; the positive surgical margin rates in pT2 cancers were 12.2% and 11.7% (P = 0.70). For urinary continence, 41% of patients having RRP and 68.9% of those having RALP were continent at catheter removal (P < 0.001). The 12-month continence rates were 88% after RRP and 97% after RALP (P = 0.01), with the mean time to continence being 75 and 25 days (P < 0.001), respectively. At the 12-month follow-up, 20 of 41 patients having bilateral nerve-sparing RRP (49%) and 52 of 64 having bilateral nerve-sparing RALP (81%) (P < 0.001) had recovery of erectile function. CONCLUSIONS: RALP offers better results than RRP in terms of urinary continence and erectile function recovery, with similar positive surgical margin rates. PMID- 19281469 TI - Does urodynamic verification of overactive bladder determine treatment success? Results from a randomized placebo-controlled study. AB - OBJECTIVE: To determine whether subjects with or with no detrusor overactivity (DO) determined by urodynamic assessment respond differently to treatment with the antimuscarinic agent tolterodine (extended release formulation, ER). SUBJECTS AND METHODS: Adult subjects with urinary frequency (average >or=8 voids/24 h) and urgency with or without urgency urinary incontinence (UUI) underwent urodynamic assessment and were stratified according to whether they had DO (positive urodynamics) or not (negative urodynamics). Subjects in each urodynamic stratum were randomized to receive tolterodine-ER (4 mg once daily) or placebo for 12 weeks. Diary cards were completed for 7 days before each study visit (at baseline, week 4, and week 12). The volume per void was recorded for 3 of the 7 days. RESULTS: The difference between the positive and negative urodynamic groups in mean change in volume voided between baseline and 12 weeks was 5.38 mL (95% CI, -93 mL to +15.71 mL). This difference is within the pre-stipulated range defined for equivalence (+/-20 mL, P = 0.31). There was also no significant difference in the change from baseline to 12 weeks between the urodynamics groups in mean number of voids per day or UUI episodes. However, there was significant improvement in the treatment group compared with the placebo group, in the number of voids per 24 h (P = 0.003) and in the mean change in volume voided (P = 0.03), from baseline to 12 weeks, but not in UUI episodes (P = 0.35). CONCLUSIONS: Urodynamics status could not predict treatment outcomes between patients treated with tolterodine-ER or placebo. The results add support to evidence suggesting that urodynamic assessment is not a prerequisite for the treatment of overactive bladder (OAB). Therefore, we recommend that anticholinergic treatment may be initiated to patients with OAB symptoms without the need for urodynamics studies. PMID- 19281470 TI - More pro-activity is required in the battle against obesity in men. PMID- 19281471 TI - Using phylochronology to reveal cryptic population histories: review and synthesis of 29 ancient DNA studies. AB - The evolutionary history of a population involves changes in size, movements and selection pressures through time. Reconstruction of population history based on modern genetic data tends to be averaged over time or to be biased by generally reflecting only recent or extreme events, leaving many population historic processes undetected. Temporal genetic data present opportunities to reveal more complex population histories and provide important insights into what processes have influenced modern genetic diversity. Here we provide a synopsis of methods available for the analysis of ancient genetic data. We review 29 ancient DNA studies, summarizing the analytical methods and general conclusions for each study. Using the serial coalescent and a model-testing approach, we then re analyse data from two species represented by these data sets in a common interpretive framework. Our analyses show that phylochronologic data can reveal more about population history than modern data alone, thus revealing 'cryptic' population processes, and enable us to determine whether simple or complex models best explain the data. Our re-analyses point to the need for novel methods that consider gene flow, multiple populations and population size in reconstruction of population history. We conclude that population genetic samples over large temporal and geographical scales, when analysed using more complex models and the serial coalescent, are critical to understand past population dynamics and provide important tools for reconstructing the evolutionary process. PMID- 19281472 TI - Activation of the brainstem but not of the hypothalamus in hemicrania continua without autonomic symptoms. AB - A 64-year-old woman presented with a 6-month history of right-sided continuous headache, without autonomic symptoms and complete response to indomethacin. Clinical examination and structural brain imaging were normal. A diagnosis of hemicrania continua (HC) was made. We sought to determine the brain structures active during the pain in a patient who met all of the diagnostic criteria for HC with the exception of autonomic symptoms. A brain positron emission tomography study was performed during pain, and completely pain-free after indomethacin administration. Comparing the pain with pain-free states, the region of the dorsal pons was significantly activated. There was no activation in the hypothalamus, as previously reported in HC with autonomic symptoms. Although definitive conclusions can not be drawn from a single observation, the lack of autonomic symptoms along with the absence of hypothalamic activation suggests that the clinical presentation may predict the pattern of brain activation in primary headache syndromes. PMID- 19281473 TI - Cluster headache in 2-year-old Polish girl. PMID- 19281474 TI - Analysis of the relationship between age and treatment response in migraine. AB - In migraine, headache severity varies with age. As a consequence, the effectiveness of medication may also depend on a patient's age. The purpose of this study was to assess the combined effect of age and drug treatment on headache characteristics. Using data from clinical trials of sumatriptan in adolescents and adults, we show how the interaction between age and drug exposure can be parameterised as a covariate on a Markov model that describes the decline of headache severity over three clinically defined stages (no relief, relief and pain-free status). The model explains important clinical observations: (i) the rates at which the pain relief and pain-free status were attained were found to be inversely related to age; (ii) in placebo-treated patients, the mean transit time from 'no relief' to 'relief' is 3 h for young adolescents and increases to 6 h for patients aged >or= 30 years; and (iii) sumatriptan reduces the transit time to 2 h, irrespective of age. These findings indicate that the therapeutic gain over placebo increases with age. Prospective studies of antimigraine drugs should take this relationship into account when extrapolating efficacy data from adults to adolescents. PMID- 19281475 TI - Promiscuous mitochondria in Cryptococcus gattii. AB - Cryptococcus gattii is a primary pathogenic basidiomycetous yeast comprising four genotypic groups. Here we present data on two mitochondrial loci (MtLrRNA and ATP6). Two of the genotypic groups, namely amplified fragment length polymorphism (AFLP)5/VGIII and AFLP6/VGII, formed monophyletic lineages. The AFLP4/VGI genotypic group, however, possessed five different mitochondrial genotypes that did not form a monophyletic lineage. The majority of these isolates contained mitochondrial genomes that are partially identical to those found in isolates belonging to AFLP6/VGII, which is causing the ongoing and expanding Vancouver Island outbreak. Two out of four AFLP7/VGIV isolates contained an AFLP4/VGI allele of MtLrRNA. These observations are best explained by assuming a process of mitochondrial recombination. If this is true, mitochondrial recombination seems possible between cells belonging to different genotypic groups of C. gattii, especially between AFLP6/VGII or AFLP7/VGIV and AFLP4/VGI. We also have to assume that mitochondria, most likely, were transferred from cells belonging to AFLP6/VGII to AFLP4/VGI. As such a process of mitochondrial recombination is only possible after cell-cell conjugation, this may also allow the further exchange of genetic material, for example nuclear or plasmid in nature, between different genotypes of C. gattii. This may be relevant as it may provide a possible mechanism contributing to the modulation of virulence attributes of isolates, such as has been observed in the ongoing Vancouver Island outbreak of C. gattii. PMID- 19281476 TI - Direct spectroscopic (FTIR) detection of intraspecific binary contaminations in yeast cultures. AB - Fourier transform infrared spectroscopy has proved to be a good method to identify and characterize microorganisms. This technique has been proposed as a tool to determine the level of contamination in binary mixtures of strains belonging to different species and even to diverse kingdoms, showing a good linear relationship between spectral outputs and contamination levels. The monitoring of intraspecific contamination is a critical point in both laboratory practice and industrial monitoring, but it is challenged by the difficulty to discriminate between very similar cultures belonging to the same species. In this paper we considered binary intraspecific mixtures of strains belonging to three species (Saccharomyces cerevisiae, Debaryomyces hansenii and Rhodotorula minuta). Results showed that contaminated and pure cultures can be discriminated on the basis of their infrared spectra and that different spectral areas respond to the contamination according to the species under test. Moreover, some spectral areas change linearly with the increase of contaminants, giving the possibility of using this procedure for preliminary estimations of the contamination in addition to the even more important opportunity to indicate the presence of contaminants of the same species at low levels in fermentation cultures. PMID- 19281477 TI - Nonmicrobial aerobic methane emission from poplar shoot cultures under low-light conditions. AB - The aerobic formation of methane in plants has been reported previously, but has been questioned by a number of researchers. Recently, isotopic evidence demonstrated that ultraviolet irradiation and heating lead to photochemical or thermal aerobic methane formation mainly from plant pectin in the absence of microbial methane production. However, the origin of aerobic methane formation from plant material observed under low temperature and low-light/dark conditions is still unclear. Here we show that Grey poplar (Populus * canescens, syn. Populus tremula * Populus alba) plants derived from cell cultures under sterile conditions released 13C-labeled methane under low-light conditions after feeding the plants with 13CO2. Molecular biological analysis proved the absence of any microbial contamination with known methanogenic microorganisms and ruled out the possibility that methane emission from our poplar shoot cultures under aerobic low-light/dark and ambient temperature conditions could be of microbial origin. The CH4 release rates in our experiment were in the range of 0.16-0.7 ng g-1 DW h 1, adding evidence to the growing opinion that the quantitative role of aerobic methane emissions from plants in the global methane budget, at least from cold temperate or boreal regions, is only of minor importance. PMID- 19281478 TI - Increased proline loading to phloem and its effects on nitrogen uptake and assimilation in water-stressed white clover (Trifolium repens). AB - The aim of this study was to investigate the physiological significance of increased proline loading to phloem caused by water-deficit stress in relation to nitrogen (N) uptake and assimilation. N uptake and N assimilation were quantified by 15N tracing in well-watered (control) and water deficit-stressed white clover (Trifolium repens). De novo proline synthesis and proline loading to the phloem were also compared between treatments. The relationships among proline concentrations in phloem exudates, N uptake, and assimilation of newly absorbed N were assessed. The newly synthesized proline in the phloem exudates increased rapidly after 3 d of water deficit. The water-deficit treatment significantly reduced the maximum nitrate reductase activity (NRA), and also attenuated de novo synthesis of amino acids and proteins in the roots. The increase in proline concentrations in phloem exudates was closely related to reductions in NRA in the roots, N uptake, and the assimilation of newly absorbed N. The accumulation of proline induced in roots by exogenous proline and NH4Cl treatments was closely associated with the decrease in NRA. These results indicate that increased proline transport to roots via phloem caused by water deficit has a significant influence on the down-regulation of N uptake and the assimilation of newly absorbed N. PMID- 19281479 TI - Impact of laryngeal mask airway cuff pressures on the incidence of sore throat in children. AB - BACKGROUND: Hyperinflation of laryngeal mask airway cuffs can cause harm to the upper airway mainly by exerting high pressures on pharyngeal and laryngeal structures thus impairing mucosal perfusion. Although cuff manometers can be used to guide the monitoring of cuff pressures, their use is not routine in many institutions. In a prospective audit, we assessed the incidence of sore throat following day-case-surgery in relation to the intracuff pressure within the laryngeal mask airway. METHODS: Four hundred children (3-21 years) were consecutively included in this study. The laryngeal mask airway was inflated as deemed necessary by the attending anesthetist. Cuff pressures were measured using a calibrated cuff manometer (Portex Limited, Hythe, Kent, UK, 0-120 cm H2O, pressures exceeding the measurement range were set at 140 cm H2O for statistical purposes) at induction of anesthesia. RESULTS: Forty-five children (11.25%) developed sore throat, 32 (8%) sore neck and 17 (4.25%) sore jaw. Of those that developed sore throat, 56.5% had cuff pressures exceeding >100 cm H2O. In contrast, when cuff pressures were <40 cm H2O, there were no episodes of sore throat, whilst there was only a 4.6% occurrence of sore throat if cuff pressures were between 40-60 cm H2O. CONCLUSION: We have demonstrated that intra cuff pressure in laryngeal mask airways is closely related to the development of sore throat with higher pressures increasing its likelihood. Hence, cuff pressures should be measured routinely using a manometer to minimize the incidence of sore throat. PMID- 19281480 TI - Pulse oximetric thresholds for tonsillectomy and adenotomy in children: significance of 1-2% decline in oxyhemoglobin saturation. AB - OBJECTIVES: We aimed to establish optimal overnight pulse oximetric thresholds for determining the indication of tonsillectomy and adenotomy (TA) in children by revising the definition of 'desaturation'. METHODS: One hundred and thirty four children scheduled for TA (TA group, 5.3 +/- 1.4 years old) and 112 otherwise healthy children scheduled for elective minor surgery (control group, 5.4 +/- 1.5 years old) were enrolled into this prospective study. Data were recorded and stored every 10 s using Nellcor N-395. Desaturation/resaturation events were defined as x% change (x = 1-4) of SpO2 (oxyhemoglobin saturation by pulse oximetry) in 10 s. The desaturation/resaturation indices were calculated as events per hour of total sleeping time. For each index, a wide range of temporary thresholds was set. The optimal thresholds for TA were the ones that maximized the weighted average for sensitivity, specificity (based on whether the index improved or not after TA), and the percentage of the control children whose indices were below the threshold. RESULTS: For all the indices, the optimal thresholds that fulfilled the above condition were determined. Compared with the x = 3-4% results, the application of x = 1-2% approximately doubled the TA patients whose preoperative 'positive' indices improved after TA, with the weighted averages of 84.3-92.3% as described above. CONCLUSIONS: By defining desaturation/resaturation as a 1-2% change in SpO2 from the preceding value, children with adenotonsillar hypertrophy whose pulse oximetric indices are expected to improve after TA can be detected by pulse oximetry with relatively high sensitivity and specificity. PMID- 19281481 TI - Endometrial and cervical polyps in 22 baboons (Papio sp.), 5 cynomolgus macaques (Macaca fascicularis) and one marmoset (Callithrix jacchus). AB - BACKGROUND: Endometrial and cervical polyps are masses of endometrium or cervical epithelium that bulge into the uterine or cervical lumen. The physiopathology and contributing factors of endometrial polyps development are still unknown. METHODS: Clinical and pathology records of 28 non-human primates with histologically confirmed endometrial and cervical polyps were reviewed. Twenty one baboons with endometrial polyps were evaluated for age at diagnosis, body weight, menstrual cycle length, presence of endometriosis and adenomyosis and number of offspring, cesarean sections, and stillbirths. RESULTS: Endometrial polyps in baboons were associated with increased age, decreased menstrual cycle lengths, endometriosis, and decreased parity. No differences were found for weight, adenomyosis, or number of cesarean sections or stillbirths. CONCLUSIONS: Baboons are a promising model for the study of endometrial polyps because of their similarity to humans in both the development of endometrial polyps and association of many of the same risk factors. PMID- 19281483 TI - High Definition Oscillometry: a novel technique for non-invasive blood pressure monitoring in the cynomolgus monkey (Macaca fascicularis). AB - BACKGROUND: Current approaches for accurate blood pressure determination rely predominantly on invasive techniques. High Definition Oscillometry (HDO) was evaluated as a potential non-invasive approach for accurate blood pressure recordings in cynomolgus monkeys. METHODS: In conscious animals, systolic, diastolic, mean arterial blood pressure (MAP) and pulse/minute were determined 15 times within approx. 9 minutes per individual. This session was performed during 3 consecutive days. Anaesthesia induced hypotension was controlled simultaneously with HDO and telemetry as reference. RESULTS: Repeated measurements were highly reproducible. After procedural habituation, mean MAP was 96.2 +/- 13.7 mmHg in males and 86.9 +/- 4.3 mmHg in females. Mean intraindividual coefficients of variation ranged between 10.8% and 2.4% depending on the session and parameter. Values determined by HDO corresponded to those reported for invasive techniques. CONCLUSION: Our results demonstrate, using telemetry as reference, the accuracy of HDO-based non-invasive blood pressure measurements in macaques to detect drug related cardiovascular changes. PMID- 19281482 TI - Fatal acute Chagas disease in a chimpanzee. AB - BACKGROUND: Chagas disease (CD) or American trypanosomiasis is caused by a hemoflagellate protozoan, Trypanosoma cruzi. This organism has been isolated from more than 100 mammalian species and several insect vectors demonstrating a wide host distribution and low host specificity. METHODS: A 23-year-old male chimpanzee died acutely and a complete necropsy was performed to evaluate gross and microscopic pathologic changes. After observation of trypanosomal amastigotes in the myocardium, PCR and immunohistochemistry was employed to confirm the diagnosis of T. cruzi. RESULTS: Gross findings were consistent with mild congestive heart failure. Microscopic findings included multifocal myocardial necrosis associated with severe lymphocytic to mixed inflammatory infiltrates, edema, and mild chronic interstitial fibrosis. Multifocal intracytoplasmic amastigotes morphologically consistent with T. cruzi were observed in cardiac myofibers. Trypanosoma cruzi was confirmed by PCR and immunohistochemistry. CONCLUSION: We report, to the best of our knowledge, the first fatal spontaneous case of T. cruzi infection in a chimpanzee. PMID- 19281484 TI - Adipose tissue remodeling in lipedema: adipocyte death and concurrent regeneration. AB - Lipedema is a disease with unknown etiology presenting as bilateral and symmetric enlargement of the lower extremities due to subcutaneous deposition of the adipose tissue. Here we describe the histopathological features of the lipedema tissue and nonaffected adipose tissue obtained from a typical patient with severe lipedema. Immunohistochemical analyses indicated degenerative and regenerative changes of the lipedema tissue, characterized by crown-like structures (necrotizing adipocytes surrounded by infiltrating CD68+ macrophages; a feature commonly seen in obese adipose tissue) and proliferation of adipose-derived stem/progenitor/stromal cells (Ki67+CD34+ cells), respectively. These findings suggested increased adipogenesis in the lipedema tissue, which may further lead to hypoxia similar to that seen in obesity, resulting in adipocyte necrosis and macrophage recruitment. The confinement to the lower extremities and the difference from systemic obesity warrants further elucidation in future studies. PMID- 19281485 TI - Multinucleate giant cells in neurofibromas: a clue to the diagnosis of neurofibromatosis. AB - BACKGROUND: A 54-year-old African-American male patient underwent removal of several cutaneous neurofibromas. Histopathologic examination revealed a nonencapsulated, haphazardly arranged proliferation of slender spindle cells in a myxoid stroma consistent with neurofibroma. Interestingly, each specimen exhibited numerous, large, multinucleate giant cells with nuclei arranged in a wreath-like or linear pattern. Immunoperoxidase staining revealed these cells to be negative with S-100 and CD-34. It was subsequently learned that this patient has neurofibromatosis type 1 (NF1). METHODS: A retrospective review of all lesions coded as neurofibroma from our institution between 1 June 2006 and 28 August 2006 was performed. RESULTS: Biopsies of 53 cutaneous neurofibromas from 51 patients were reviewed. In these, multinucleate giant cells were present in only three (5.7%), all in patients with single lesions. CONCLUSIONS: Although the incidence of floret-like giant cells in neurofibromas of patients with NF1 is unknown, our findings suggest that the presence of these cells might be a clue to the presence of NF1. PMID- 19281486 TI - Two cutaneous malignant melanomas at the same anatomic site: a case report with molecular evaluation. AB - Patients who have had malignant melanoma are at an increased risk of developing a second primary melanoma compared with the general population risk of developing a first melanoma. Many of these second primary melanomas occur at a similar anatomic site as the first lesion. Determining whether a second lesion is indeed a separate primary vs. a metastasis or locoregional recurrence can be very difficult histologically. We report the case of a patient who developed a second melanoma, 2 years after the initial diagnosis, within 3 cm of the site of the original lesion. Because of distinct histomorphologic features, the second lesion was favored to be a separate primary. However, because of the nearly identical anatomic location, molecular testing for loss of heterozygosity and BRAF mutation was performed to help further make this distinction. The first lesion was found to have loss of heterozygosity and a BRAF mutation that were not present in the second lesion. While these tests cannot elucidate the true molecular origin of these lesions, they provide a useful clinical tool to assess whether a second lesion should be treated as a recurrence or as a separate lesion with unique biologic potential. PMID- 19281487 TI - Sequential processing of hepatitis C virus core protein by host cell signal peptidase and signal peptide peptidase: a reassessment. AB - Hepatitis C virus (HCV) core protein is believed to play critical roles in the virus morphogenesis and pathogenesis. In HCV polyprotein, core protein terminates with a signal peptide followed by E1 envelope protein. It has remained unclear whether cleavage by host cell signal peptidase (SP) at the core-E1 junction to generate the complete form of core protein, which is anchored in the endoplasmic reticulum membrane, is absolutely required for cleavage within the signal peptide by host cell signal peptide peptidase (SPP) to liberate the mature form of core protein, which is then free for trafficking to lipid droplets. In this study, the possible sources of disagreement in published reports have been examined, and we conclude that a product generated upon inhibition of SP-catalysed cleavage at the core-E1 junction in heterologous expression systems was incorrectly identified as mature core protein. Moreover, inhibition of this cleavage in the most relevant model of human hepatoma cells replicating a full-length HCV genome was shown to abolish interaction of core protein with lipid droplets and production of infectious progeny virus. These results firmly establish that SPP-catalysed liberation of mature core protein is absolutely dependent on prior cleavage by SP at the correct core-E1 site to generate the complete form of core protein, consistent with this obligatory order of processing playing a role in HCV infectious cycle. PMID- 19281488 TI - Entire genome sequences of two new HCV subtypes, 6r and 6s, and characterization of unique HVR1 variation patterns within genotype 6. AB - Hepatitis C virus genotype 6 currently contains 21 recognized subtypes, 6a-6u, for which 6r and 6s lack complete genome sequences. In this study, we entirely sequenced variants QC245 and QC66 from Cambodian immigrants in Canada representing subtypes 6r and 6s, respectively. The two genomes shared 75.3% nucleotide similarities to each other and 72.0-82.9% to 21 reference sequences representing subtypes 6a-6q, 6t-6u and variants km41 and gz52557. QC66 and QC245 displayed genome lengths of 9473 and 9450 nt and each contained a single open reading frame of 9051 nt. In 10 protein encoding regions QC245 and QC66 shared common sizes with TV249/6t and 537796/6l isolates, respectively. Phylogenetic analyses demonstrated that QC245 was more closely related to subtype 6f, but both QC66 and QC245 were subtypically different from all other genotype 6 subtypes. Our full-length sequence data confirmed the status of subtype 6r and 6s within genotype 6. Analysis of partial sequences revealed seven 6t and two 6s isolates that were all isolated from Cambodian immigrants. Analysis of the hypervariable region 1 sequences of 81 genotype 6 variants revealed two unique patterns of variation. First, most variants showed an amino acid deletion at the 4th position and second, many contained a basic residue at the 7th position. Possible roles of these two variation patterns are further discussed. PMID- 19281489 TI - Role of RNA-binding proteins in mammalian spermatogenesis. AB - Spermatogenesis is a cell differentiation programme that allows a normally dividing diploid cell to become haploid and to acquire the morphological characteristics required to reach and to fertilize the female gamete. Many of the steps involved in this differentiation programme necessitate profound modifications of the genome, rendering it unable to play its template role for the synthesis of mRNAs. Therefore, de novo transcription is not a continuous process during germ cell differentiation and many mRNAs need to be synthesized and stored at specific times to be available during the transcriptionally inactive stages of spermatogenesis. Germ cells express high levels of RNA-binding proteins that assist these post-transcriptional events. The generation of mouse knockout models has highlighted the essential role played by many of these RNA binding proteins for the correct progress of spermatogenesis and for the formation of a fertile male gamete. Herein, we review the major findings on the role of RNA-binding proteins in mammalian spermatogenesis. PMID- 19281490 TI - Improvement of semen quality in an infertile man with 21-hydroxylase deficiency, suppressed serum gonadotropins and testicular adrenal rest tumours. AB - Here, we report improvement of semen quality in a 30-year-old man with congenital adrenal hyperplasia (CAH) because of 21-hydroxylase deficiency, bilateral testicular adrenal rest tumours (TART) and a 1.5-year infertility history. His adrenal substitution therapy was changed from hydrocortisone 10 mg once daily to 10 mg three times daily in combination with dexamethasone, 0.1 mg once daily. Upon this change, the testicular tumours showed regression and his sperm concentration increased from 0.1 to 98 mio/mL and total sperm count from 0.64 to 392 mio. sp., total number of motile spermatozoa increased from 0.23 to 258.72 mio. The total number of morphologically normal spermatozoa increased from 0.01 to 19.6 mio. sp. The patient reported improved well being and did not develop signs of overtreatment. A non-optimal medical substitution therapy of this CAH patient is the most likely explanation for the presence of TART and disturbed reproductive hormones levels, leading to impaired semen quality. Optimizing the medical treatment may at least in some cases improve fecundity. PMID- 19281491 TI - Semen quality, reproductive hormones and fertility of men operated for hypospadias. AB - The testicular function of men previously operated for hypospadias has been sparsely investigated. Therefore, we investigated semen quality and reproductive hormones of 92 men with isolated hypospadias (IH) and 20 with hypospadias and additional genital disorders (HAGD) and compared with similar results from young men from the general Danish population. All participants lived the Copenhagen area of Denmark. Additionally, fertility information on 1083 men registered as operated for hypospadias was retrieved from national registries. The semen quality of men with IH did not differ from controls, but was reduced in men with HAGD. Median values for IH and HAGD were, respectively: sperm concentration 52 and 32 million (mill)/mL (p = 0.02), total sperm counts 173 and 101 mill (p = 0.03), motile spermatozoa 70 and 58% (p = 0.007) and morphological normal spermatozoa 9 and 4% (p = 0.004). Men with IH had a slight increase in follicle stimulating hormone and luteinizing hormone levels, whereas men with HAGD had more pronounced disturbances. 24.0% of the 1083 men operated for hypospadias were registered as fathers to at least one child, whereas the corresponding number in the general age-matched population was 29.4% (p < 0.01). In conclusion, the majority of men with IH had normal semen quality, whereas it was reduced for men with HAGD. However, reproductive hormone levels indicated a subtle impairment of testicular function also in men with IH. An observed lower number of fathers among men with hypospadias may be because of psychosocial aspects, sexual dysfunction or reduced semen quality or a combination of these factors. Our results should be reassuring for patients with mild forms of IH and their relatives. They can be informed that hypospadias in such cases is not generally associated with poor semen quality. Particularly among patients with HAGD, several may, however, need fertility treatment to reproduce. PMID- 19281492 TI - alpha1-antitrypsin prevents polymorphonuclear leucocyte-elastase effects on spermatozoa quality. AB - Elevated levels of polymorphonuclear leucocyte (PMN)-derived elastase, which is suggested as marker for inflammations in the male genital tract, correlate well with spermatozoa deterioration. PMN elastase caused a time- and concentration dependent (up to a elastase concentration of 0.5 microg/mL) externalization of phosphatidylserine and intercalation of propidium iodide on human spermatozoa. There are apparently a limited number of target sites for elastase on spermatozoa surface, because the further enhancement of elastase amount did not fasten alterations in spermatozoa parameters. Analysis of flow cytometry data revealed that most spermatozoa were in a necrotic state after an exposure with elastase for 22 h. Some apoptotic cells were only detected at shorter incubation periods. Seminal plasma prevented in a concentration-dependent manner the PMN elastase mediated loss of vitality of spermatozoa. We detected by blotting techniques large amounts of alpha(1)-antitrypsin in seminal plasma. This antiproteinase is known to inactivate elastase at inflammatory sites. Increasing concentrations of alpha(1)-antitrypsin prevented gradually spermatozoa deterioration induced by elastase. Thus, alpha(1)-antitrypsin contributes to an efficient protease/antiproteinase balance in seminal plasma. A disturbed balance will promote the development of chronic inflammations which can also be the reason for male infertility problems. PMID- 19281493 TI - Identification of fall risk factors in older adult emergency department patients. AB - OBJECTIVES: Falls represent an increasingly frequent source of injury among older adults. Identification of fall risk factors in geriatric patients may permit the effective utilization of scarce preventative resources. The objective of this study was to identify independent risk factors associated with an increased 6 month fall risk in community-dwelling older adults discharged from the emergency department (ED). METHODS: This was a prospective observational study with a convenience sampling of noninstitutionalized elders presenting to an urban teaching hospital ED who did not require hospital admission. Interviews were conducted to determine the presence of fall risk factors previously described in non-ED populations. Subjects were followed monthly for 6 months through postcard or telephone contact to identify subsequent falls. Univariate and Cox regression analysis were used to determine the association of risk factors with 6-month fall incidence. RESULTS: A total of 263 patients completed the survey, and 161 (61%) completed the entire 6 months of follow-up. Among the 263 enrolled, 39% reported a fall in the preceding year, including 15% with more than one fall and 22% with injurious falls. Among those completing the 6 months of follow-up, 14% reported at least one fall. Cox regression analysis identified four factors associated with falls during the 6-month follow-up: nonhealing foot sores (hazard ratio [HR] = 3.71, 95% confidence interval [CI] = 1.73 to 7.95), a prior fall history (HR = 2.62, 95% CI = 1.32 to 5.18), inability to cut one's own toenails (HR = 2.04, 95% CI = 1.04 to 4.01), and self-reported depression (HR = 1.72, 95% CI = 0.83 to 3.55). CONCLUSIONS: Falls, recurrent falls, and injurious falls in community dwelling elder ED patients being evaluated for non-fall-related complaints occur at least as frequently as in previously described outpatient cohorts. Nonhealing foot sores, self-reported depression, not clipping one's own toenails, and previous falls are all associated with falls after ED discharge. PMID- 19281494 TI - Statin therapy is associated with decreased mortality in patients with infection. AB - OBJECTIVES: The objective was to investigate the association between statin therapy and mortality in emergency department (ED) patients with suspected infection. METHODS: A secondary analysis of a prospective, observational cohort study was conducted at an urban, academic ED with approximately 50,000 annual visits. Data were collected between December 2003 and September 2004. Inclusion criteria consisted of age > or = 18 years, clinical suspicion of infection, and hospital admission. Patients were divided by those receiving statin therapy and those not receiving statins while hospitalized. Medication data were collected from an inpatient pharmacy database. Comparisons were conducted with Fisher's exact test or Wilcoxon rank sum test. To adjust for baseline differences, multivariable logistic regression analysis controlling for gender, severity of illness (Mortality in Emergency Department Sepsis [MEDS] score), Charlson Comorbidity Index, and duration of statin therapy was performed. RESULTS: Of 2,132 patients with suspected infection, 2,036 (95%) had interpretable pharmacy data and were analyzed. The cohort had a median age of 61 years (interquartile range [IQR] = 46-78 years) and a mortality of 3.9% (95% confidence interval [CI] = 3.1% to 4.8%). Patients who received statins (n = 474) had a lower unadjusted crude mortality (1.9%; 95% CI = 0.6% to 3.3%) compared to those who did not (4.5%; 95% CI = 3.4% to 5.4%; p or =10 feet occurred in 29.3% of cases for which height of the fall/jump was recorded. Boys had significantly higher odds of falling or jumping from a height of > or =10 ft than girls, and children 10 to 19 years old also had significantly higher odds of falling or jumping from a height of > or =10 feet, compared to those 9 years old and younger. The odds of hospitalization were tripled if the patient fell or jumped from > or =10 feet and nearly tripled if the patient sustained a fracture. CONCLUSIONS: This study examined tree house related injuries on a national level. Tree house safety deserves special attention because of the potential for serious injury or death due to falls from great heights, as well as the absence of national or regional safety standards. The authors provide safety and prevention recommendations based on the successful standards developed for playground equipment. PMID- 19281498 TI - Aging, IGF-1, and diet. PMID- 19281499 TI - Demographic characteristics of patients with severe neuropathic pain secondary to failed back surgery syndrome. AB - BACKGROUND: Neuropathic pain commonly affects the back and legs and is associated with severe disability and psychological illness. It is unclear how patients with predominantly neuropathic pain due to failed back surgery syndrome (FBSS) compare with patients with other chronic pain conditions. AIMS: To present data on characteristics associated with FBSS patients compared with those with complex regional pain syndrome, rheumatoid and osteoarthritis, and fibromyalgia. METHODS: The PROCESS (Prospective Randomized Controlled Multicenter Trial of the Effectiveness of Spinal Cord Stimulation, ISRCTN 77527324) trial randomized 100 patients to spinal cord stimulation (n = 52) plus conventional medical management (CMM) or CMM alone (n = 48). Baseline patient parameters included age, sex, time since last surgery, employment status, pain location and severity (visual analogue scale), health-related quality of life (HRQoL), level of disability, medication, and nondrug therapies. Reference population data was drawn from the literature. RESULTS: At baseline, patients in the PROCESS study had a similar age and gender profile compared with other conditions. PROCESS patients suffered from greater leg pain and had lower HRQoL. PROCESS patients treatment cost was higher and they commonly took opioids, while antidepressants and nonsteroidal anti inflammatory drugs were more often used for other conditions. Prior to baseline, 87% of patients had tried at least 4 different treatment modalities. CONCLUSIONS: Patients suffering from chronic pain of neuropathic origin following FBSS often fail to obtain adequate relief with conventional therapies (eg, medication, nondrug therapies) and suffer greater pain and lower HRQoL compared with patients with other chronic pain conditions. Neuropathic FBSS patients may require alternative and possibly more (cost-) effective treatments, which should be considered earlier in their therapeutic management. PMID- 19281500 TI - Differences between smooth pursuit and optokinetic eye movements using limited lifetime dot stimulation: a functional magnetic resonance imaging study. AB - In this study, we examined possible differences in brain activation between smooth pursuit and optokinetic reflexive (OKR) eye movements using functional magnetic resonance imaging (fMRI). Eighteen healthy subjects performed two different eye movement paradigms. In the first paradigm, smooth pursuit eye movements were evoked by a single moving dot. In the second paradigm, optokinetic eye movements without a foveal smooth pursuit component were evoked by a moving pattern of multiple dots with a limited lifetime. As expected, the two eye movement systems show overlapping pathways, but the direct comparison of the activation patterns between the two experiments showed that the frontal eye field, MT/V5 and cerebellar area VI appear to be more activated during smooth pursuit than during optokinetic eye movements. These results showed that the smooth pursuit and optokinetic eye movement systems can be differentiated with fMRI using limited lifetime dots as an effective OKR stimulus. PMID- 19281501 TI - Assessment of infrazygomatic bone depth for mini-screw insertion. AB - OBJECTIVE: To investigate the bone depth at the infrazygomatic crest with regard to orthodontic mini-screw insertion. MATERIAL AND METHODS: Twenty-nine adult human dry skulls were imaged using CBCT technology, slice data were generated and multiple measurements were undertaken at three sites associated with the infrazygomatic crest and five different measurement levels. The data were analyzed using intraclass correlation and repeated measures ANOVA. RESULTS: The greatest bone depth was available at, on average, 11.48+/-1.92 mm apical from the cemento-enamel junction of the maxillary first molar and decreased rapidly further apically. Maximum bone depth (7.05+/-3.7 mm) was present at the lowest measurement level. However, here, insufficient clearance to the molar roots was present. Both the measurement site and the level at which the measurements were conducted had a significant impact on bone depth. CONCLUSIONS: When inserting orthodontic mini-screws (6 mm or longer) into the infrazygomatic crest while staying clear of the molar roots perforation of the maxillary sinus or the nasal cavity can be expected, but bone depth varies considerably between individuals. PMID- 19281502 TI - Effect of repeated torque/mechanical loading cycles on two different abutment types in implants with internal tapered connections: an in vitro study. AB - OBJECTIVES: Internal tapered connections were developed to improve biomechanical properties and to reduce mechanical problems found in other implant connection systems. The purpose of this study was to evaluate the effects of mechanical loading and repeated insertion/removal cycles on the torque loss of abutments with internal tapered connections. MATERIAL AND METHODS: Sixty-eight conical implants and 68 abutments of two types were used. They were divided into four groups: groups 1 and 3 received solid abutments, and groups 2 and 4 received two piece abutments. In groups 1 and 2, abutments were simply installed and uninstalled; torque-in and torque-out values were measured. In groups 3 and 4, abutments were installed, mechanically loaded and uninstalled; torque-in and torque-out values were measured. Under mechanical loading, two-piece abutments were frictionally locked into the implant; thus, data of group 4 were catalogued under two subgroups (4a: torque-out value necessary to loosen the fixation screw; 4b: torque-out value necessary to remove the abutment from the implant). Ten insertion/removal cycles were performed for every implant/abutment assembly. Data were analyzed with a mixed linear model (P< or =0.05). RESULTS: Torque loss was higher in groups 4a and 2 (over 30% loss), followed by group 1 (10.5% loss), group 3 (5.4% loss) and group 4b (39% torque gain). All the results were significantly different. As the number of insertion/removal cycles increased, removal torques tended to be lower. It was concluded that mechanical loading increased removal torque of loaded abutments in comparison with unloaded abutments, and removal torque values tended to decrease as the number of insertion/removal cycles increased. PMID- 19281503 TI - Soft tissues healing at immediate transmucosal implants placed into molar extraction sites with buccal self-contained dehiscences. A 12-month controlled clinical trial. AB - AIM: To assess soft tissues healing at immediate transmucosal implants placed into molar extraction sites with buccal self-contained dehiscences. MATERIAL AND METHODS: For this 12-month controlled clinical trial, 15 subjects received immediate transmucosal tapered-effect (TE) implants placed in molar extraction sockets displaying a buccal bone dehiscence (test sites) with a height and a width of > or =3 mm, respectively. Peri-implant marginal defects were treated according to the principles of Guided Bone Regeneration (GBR) by means of deproteinized bovine bone mineral particles in conjunction with a bioresorbable collagen membrane. Fifteen subjects received implants in healed molar sites (control sites) with intact buccal alveolar walls following tooth extraction. In total, 30 TE implants with an endosseous diameter of 4.8 mm and a shoulder diameter of 6.5 mm were used. Flaps were repositioned and sutured, allowing non submerged, transmucosal soft tissues healing. At the 12-month follow-up, pocket probing depths (PPD) and clinical attachment levels (CAL) were compared between implants placed in the test and the control sites, respectively. RESULTS: All subjects completed the 12-month follow-up period. All implants healed uneventfully, yielding a survival rate of 100%. After 12 months, statistically significantly higher (P<0.05) PPD and CAL values were recorded around implants placed in the test sites compared with those placed in the control sites. CONCLUSIONS: The findings of this controlled clinical trial showed that healing following immediate transmucosal implant installation in molar extraction sites with wide and shallow buccal dehiscences yielded less favorable outcomes compared with those of implants placed in healed sites, and resulted in lack of 'complete' osseointegration. PMID- 19281504 TI - Mechanical properties of the Schneiderian membrane in vitro. AB - OBJECTIVES: Perforation of the Schneiderian membrane (maxillary sinus mucosa) is a common complication of maxillary sinus graft procedures. Membrane perforation increases the chance of postoperative sinusitis and endangers graft as well as implant survival. The aim of the present study was to explore the mechanical properties of the Schneiderian membrane. MATERIAL AND METHODS: Three test methods were performed on sinus specimen of 20 fresh human cadavers: one- and two dimensional membrane elongation as far as perforation, as well as membrane detachment from the adherent bone. RESULTS: Perforation of the Schneiderian membrane (mean thickness: 90 mum) occurred at a mean tension of 7.3 N/mm(2). The membrane could be stretched to 132.6% of its original size in one-dimensional elongation, and to 124.7% in two-dimensional elongation. Thicker membranes demonstrated significantly higher load limits (P<0.001). The mean modulus of elasticity accounted 0.058 GPa, the mean adhesion force between sinus membrane and bone surface was 0.05 N/mm. CONCLUSIONS: Respecting the mechanical properties of the Schneiderian membrane may help reducing the complication rates and thus patient morbidity in minimally invasive maxillary sinus floor elevation. PMID- 19281508 TI - Teaching and assessment of implant dentistry in university education. Introduction. PMID- 19281509 TI - A survey on undergraduate implant dentistry education in Europe. AB - INTRODUCTION: The aim of the survey was to assess the status of implant dentistry education and addressed various aspects related to competence level, practical implementation and barriers for further development in the field. MATERIALS AND METHODS: An e-mail survey was performed amongst 73 opinion leaders from 18 European countries invited to the Association for Dental Education in Europe (ADEE) workshop on implant dentistry. RESULTS: Forty-nine surveys were returned (67%) and it was found that theoretical and pre-clinical courses to an average of 36 h are given to undergraduates; 70% reported that students assist or treat patients with prosthetics; 53% reported that students assist with surgery and only 5% is operating patients. In 23% of the schools optional undergraduate courses are available and 90% offer postgraduate training. Barriers for including prosthetics and surgery are lack of time, funding or staff. Partial restorations, including surgery, in the posterior regions may be provided by dentists after attendance at additional courses but complex treatments should be limited to specialists. CONCLUSION: This survey confirms that implant dentistry is part of the undergraduate curriculum, albeit with a disparity in time. Whereas implant dentistry is an important part of clinical practice, coverage in the curriculum is limited and when compared with 10 years ago, even stagnating. Priorities within the curriculum should be evaluated depending on demands and treatment needs of the population. To optimise education, learning guidelines should be developed, based on the expected competencies for practicing dentists. Undergraduate education may start the process that must continue through all levels of education, including the postgraduate level. PMID- 19281510 TI - Teaching and assessment of implant dentistry in undergraduate and postgraduate education: a European consensus. AB - This paper constitutes a summary of the consensus documents agreed at the First European Workshop on Implant Dentistry University Education held in Prague on 19 22 June 2008. Implant dentistry is becoming increasingly important treatment alternative for the restoration of missing teeth, as patients expectations and demands increase. Furthermore, implant related complications such as peri implantitis are presenting more frequently in the dental surgery. This consensus paper recommends that implant dentistry should be an integral part of the undergraduate curriculum. Whilst few schools will achieve student competence in the surgical placement of implants this should not preclude the inclusion of the fundamental principles of implant dentistry in the undergraduate curriculum such as the evidence base for their use, indications and contraindications and treatment of the complications that may arise. The consensus paper sets out the rationale for the introduction of implant dentistry in the dental curriculum and the knowledge base for an undergraduate programme in the subject. It lists the competencies that might be sought without expectations of surgical placement of implants at this stage and the assessment methods that might be employed. The paper also addresses the competencies and educational pathways for postgraduate education in implant dentistry. PMID- 19281511 TI - The rationale for the introduction of implant dentistry into the dental curriculum. AB - This paper provides arguments for the introduction of implant dentistry into the undergraduate curriculum. The survival of teeth is very high when disease is diagnosed and treated properly and maintenance is taken care of. Nevertheless, tooth replacements by fixed and removable prostheses are highly prevalent. It is expected that dentists will face a dramatically increased need to care for elderly patients and partially edentulous patients. Hence, the demand for implant reconstructions will be substantial and more appropriately trained and competent health professionals will be needed. Increasing demands of the patient regarding aesthetics and function will influence the demands for implant therapy. The improvement of oral function and subjective chewing comfort, the preservation of tooth structures or existing reconstructions and the replacement of missing, strategically important teeth are major indications for implant placement. From both a biological and an economical point of view, the single tooth replacement with an implant is the first choice in situations with no or minimally restored neighbouring teeth compared with conventional bridgework. Stability of full dentures represent a major problem especially for the mandible. It is well documented that placement of two implants supporting an overdenture substantially improve chewing capacity, increase quality of life and is a simple and cost effective treatment thus rendering such treatment a 'standard of care' procedure. There is no doubt that dental students should learn to incorporate the indication of oral implants in their overall treatment planning. Therefore, they will have to understand the basic aspects of healing and tissue integration, basic biomechanical and material science principles as well as surgical and prosthetic techniques. They will have to be able to monitor continuously the peri-implant tissues, render appropriate supportive therapy and cope with biological and technical complications. While it is evident that the surgical procedure per se may require additional competence, the remainder of the aspects mentioned should be taught in the dental curriculum. This should include the attribution of responsibility for maintenance of implants and handling of biological and technical complications. Moreover, it is desirable to include the surgical technique for implant placement for 'straightforward' cases into the dental curriculum. The levels and limitations to which the various aspects of implant dentistry and related skills are to be taught are determined by the academic community. Obviously, ethical and legal aspects of implant dentistry should not be forgotten. PMID- 19281512 TI - Theoretical knowledge in implant dentistry for undergraduate students. AB - Implant therapy has evolved into an important part of daily dental practice. Appropriate knowledge of diagnostic and therapeutic options with dental implant therapy is, therefore, mandatory for dental students. The present consensus paper describes the theoretical knowledge as a foundation to the teaching of implant dentistry at the undergraduate level. Students need a solid basic knowledge about biological prerequisites and clinical procedures leading to successful implant treatment and, in particular, an understanding of the importance of embedding implants into the overall treatment concept. Among others this includes aspects of bone and soft tissue integration of dental implants, as well as aspects of materials that are used in implant dentistry. The students should also be able to differentiate between low, medium and high-risk situations, which assumes that they have knowledge about a proper clinical examination. Furthermore, the students need to be able to inform the patient about the different treatment options and their advantages and disadvantages. Frequently, a choice has to be made between an FDP anchored on teeth and an implant-borne reconstruction. This is highly influenced by the long-term prognosis of the different treatment options. In order to perform implant placement in uncomplicated cases and to give appropriate patient information, adequate knowledge of surgical procedure and surgical complications is mandatory. Furthermore, the dentist needs to be competent in evaluating clinical situations and in advising patients about the suitability of the different options, e.g. removable or fixed reconstructions. It is possible that peri-implant tissue destruction may be a more common finding during long-term service of implant-borne reconstructions than was previously believed. The dentist needs knowledge about etiology and pathogenesis of peri implantitis and should know how to provide an effective maintenance care programme. In cases of peri-implantitis the student should be knowledgeable regarding suitable interventions. PMID- 19281513 TI - Competencies in implant therapy for the dental graduate: appropriate educational methods. AB - Modern dental education should have in on one hand the necessary education and training to make modern dentists capable of implementing population and individual preventive strategies, and on the other make them competent to satisfy current public demands to most patients. Amongst the restorative procedures most demanded by patients implant therapy has reached a clear pre-dominance. Although specialists have traditionally rendered this therapeutic procedure, in simple cases there is seldom need for extensive surgical procedures and therefore there is a clear need to introduce these studies in modern dental curricula. To facilitate the harmonisation and convergence of dental education in Europe, the Association for Dental Education in Europe and DentEd have defined a set of competencies for the graduating dentist in the EU. Using the same framework, we herein define the agreed competencies in implant dentistry, within the different domains, for both the undergraduate and the postgraduate student. Within a context of a competence-based curriculum implant dentistry should be taught as part of the major and supportive competencies described with a strong emphasis on inter-disciplinarity and integration. PMID- 19281514 TI - An outline of competencies and the appropriate postgraduate educational pathways in implant dentistry. AB - The use of dental implants has become a widely accepted and well-documented treatment option offering to both patients and dentists an alternative to traditional treatment modalities and at the same time opening a brand new area in dental postgraduate education. As such, it is necessary to define the competencies that the graduate student/dentist will need at the different levels of clinical practice in Implant Dentistry and the educational pathways that are required to convey those competencies in a structured manner. The present position paper provides an initial suggestion for the knowledge, skills and behaviour necessary for a graduate student to practice implant dentistry at the different levels of clinical complexity. An outline of the necessary competencies and structure of various levels of postgraduate university courses is provided together with different educational approaches to support them. The present paper should be evaluated as a platform for discussion for future development of postgraduate curricula in implant dentistry rather than a manual on how to design and operate such curricula. PMID- 19281515 TI - Assessment of knowledge and competencies related to implant dentistry in undergraduate and postgraduate university education. AB - Learning in academic settings is strongly related to the way the students are tested or examined. Assessment therefore must be integrated in the curriculum design, coordinated and should reflect the learning outcomes of the education. Assessment within the field of implant dentistry must fulfil four major objectives: complete and direct the learning process with feedback (formative), ensure that students are adequately prepared (summative), assess attitudes and skills such as critical thinking, reflection and self-assessment ability, and supply continuous feedback to teachers on curricular content and impact. Different assessment methods should be used to assess different levels of competencies throughout the curriculum. Various forms of written or oral assessment methodologies are applicable at earlier stages in the curriculum. At intermediate levels, interactive assessment methods, such as patient simulations (paper based or virtual) and more could encourage the necessary synthesis of several disciplines and aspects of the theoretical knowledge. At higher levels of competence, documentation of clinical proficiency by means of reflective portfolios and diaries is an appropriate assessment method with both formative and summative potential. The highest level of competence requires performance assessment using structured, objective, clinical criteria. The group strongly encourages the use of reflective forms of assessment methods which engage the students in a process of self-appraisal, identification of individual learning needs and self-directed learning. The ultimate goal of this would be to allow the student to develop a lifelong learning attitude. PMID- 19281516 TI - Gender differences among Canadian spousal caregivers at the end of life. AB - The purpose of this study was to examine gender differences in spousal caregiving at the end of life. The primary research question was to determine gender differences in caregiver strain among spousal caregivers. Secondary research questions investigated included (i) the presence of gender differences among spousal caregivers in the duration of care provided; (ii) gender differences among spousal caregivers in formal service use and unmet service needs; and (iii) whether support to care recipients in activities of daily living varied according to the gender of the spousal caregiver. The study was conducted over a 2-year period (2000-2002) in south-central Ontario, Canada. The study sample included 283 informal spousal caregivers (198 females, 85 males) each of whom were caring for a terminally ill spouse at the time they participated in a cross-sectional telephone survey. The analysis showed that females reported a significantly greater level of caregiving strain than males (t = -2.12, d.f. = 281, P = 0.035). When considering source of support in activities of daily living for the care recipient, differential assistance was noted on the basis of caregiver gender. Female caregivers had almost twice the odds of providing support in toileting related tasks than male caregivers (odds ratio (OR) = 1.98, 95% confidence interval (CI) = 1.01-3.85, P = 0.044), while male caregivers had approximately twice the odds of providing support in mobility-related tasks (OR = 0.41, 95% CI = 0.21-0.81, P = 0.011). Care recipients who had a female caregiver had lower odds of receiving support from family and friends in tasks associated with personal care (OR = 0.17, 95% CI = 0.05-0.53, P = 0.002). To address gender differences in caregiving, a realistic home-based palliative care approach must take into account the importance of informal caregivers. PMID- 19281517 TI - The effects of social support on maternal anxiety and depression after stillbirth. AB - While most births result in a live baby, stillbirth (the birth of a dead baby) occurs in nearly 1 in 110 pregnancies. This study examined whether levels of maternal anxiety and depression are lower amongst mothers who received social support after stillbirth. Using non-probability sampling, data were collected from 769 mothers residing within the USA who experienced a stillbirth within the past 18 months and for whom we have complete data. The study Maternal Observations and Memories of Stillbirth and the website http://www.momstudy.com containing the questionnaire were open in the period 8 February 2004-15 September 2005. Congruent with the family stress and coping theory, mothers of stillborn babies who perceived family support in the period after stillbirth experienced levels of anxiety and depression that were notably lower than those of their counterparts. Nurses, physicians and support groups also were important sources of support after a stillbirth; however, these sources of support alone were not statistically significant in reducing anxiety and depression in grieving mothers. Community interventions should focus on the grieving mother and her family system, including her partner and surviving children. PMID- 19281518 TI - Developing community in care homes through a relationship-centred approach. AB - Within the literature, the formation of therapeutic relationships between professionals, older people and others significant to them in their lives has been considered as central to current care philosophies. Furthermore, relationships between staff, residents and their families have emerged within the literature as fundamental to the experiences of life within the community of a care home. This paper reports part of a wider study that explored relationships between residents, families and staff. The aim of this paper is to contribute to an understanding of the factors that may be significant in the formation of relationships in care homes, and how this may support the development of community. Three case studies of care homes were undertaken using a constructivist approach. Constructivist methodology seeks to share multiple perceptions between participants with the aim of creating a joint construction. This process supported the development of shared meanings as views and ideas were shared between participants using interviews, participant observation and focus groups. The key factors influencing relationships that emerged were leadership, continuity of staff, personal philosophy of staff and contribution of residents and families. This paper suggests that considering how the style of leadership influences the organisation of care may be a useful starting point in developing community within care homes. PMID- 19281519 TI - Should invasive arteriography before carotid endarterectomy be mandatory? PMID- 19281520 TI - Strategies in infrapopliteal intervention: improving outcomes in challenging patients. PMID- 19281521 TI - Passive and active polymer coatings for intracoronary stents: novel devices to promote arterial healing. AB - Coronary stent implantation is the second great advance in the treatment of obstructive coronary artery disease since the introduction of balloon catheter angioplasty. However, in-stent restenosis (ISR) caused by neointimal hyperplasia has been a major limitation of stents, occurring in up to 30% of cases. Advances in coronary stent technology both in terms of stent design and function and especially drug-eluting stents (DES) have significantly improved the safety and efficacy of percutaneous coronary intervention (PCI) with stenting, including marked reduction in ISR. This has led to use of DES for increasingly challenging clinical and lesional subsets, with potential for increased risk of stent associated complications, especially late stent thrombosis (LST). Because restenosis and stent thrombosis are caused by multiple and often interrelated factors, ideal agents for stent coatings should inhibit thrombus formation, inflammatory reaction, and cellular proliferation, while supporting reendothelialization. To avoid undesirable effects of currently applied (durable) polymers, biocompatible, and bioabsorbable polymers as well as DES delivery systems that minimize polymer burden have been produced and tested. Bioabsorbable stents, both polymeric and metallic, have been developed to decrease potential late complications after stent implantation. Novel strategies to address some of these challenges are in various stages of research and development. In this article we outline developments in the field of passive and active stent coatings and evaluate the ongoing role of such coatings in the contemporary era of DES. PMID- 19281522 TI - Impact of bivalirudin use on outcomes in nonagenarians undergoing percutaneous coronary intervention. AB - BACKGROUND: With an aging population, nonagenarians constitute an increasing percentage of patients with coronary artery disease. The aim of this study was to determine the predictors and outcome of nonagenarians undergoing percutaneous coronary intervention (PCI) for symptomatic coronary artery disease. METHODS: From 2002 to 2007, a cohort of 171 consecutive nonagenarians underwent PCI and stent implantation in our center. Patients given bivalirudin (n = 79) during the procedure were compared to those given heparin (n = 92). In-hospital and 6-month rates of major adverse cardiovascular events (MACE) including death, myocardial infarction (MI), and target lesion revascularization were indexed. In-hospital bleedings were also indexed. RESULTS: The mean age was 92.5 +/- 2.5 years. The population was 52% female. Nearly 30% of patients had diabetes mellitus and >25% had renal failure. The mean left ventricular ejection fraction was 45%+/- 14%. The clinical presentation was an acute MI in 59% of the cases. The in-hospital rates of death and bleedings were 4.1% and 17.5%, respectively. Clinical presentations as MI or cardiogenic shock were associated with high rates of in hospital death: 19.3% and 30%, respectively. Bivalirudin use was associated with a 41.5% decrease in in-hospital bleedings. The 6-month incidence of MACE was 13.6% and was driven by death. Predictors of 6-month outcomes were clinical presentations as MI or cardiogenic shock, renal failure, and total stent length. CONCLUSION: Nonagenarians represent a particular population with a high percentage of females and a high incidence of comorbidities. In this study, we highlighted that nonagenarians have logically a worse prognosis than is reported in younger patients, with especially high rates of in-hospital bleedings. Bivalirudin use was associated with an important decrease in in-hospital bleedings; thus, it should be systematically considered in such patients to improve early outcome. PMID- 19281523 TI - The influence of morphological changes in amplatzer device on the atrial and aortic walls following transcatheter closure of atrial septal defects. AB - OBJECTIVE: This study prospectively investigated morphological changes in Amplatzer Septal Occluder (ASO) over time and the influences of these changes on the atrial and aortic walls after atrial septal defect (ASD) closure. METHODS: Between August 2005 and December 2007, 78 patients with ASD were treated with ASO devices and changes in the device shape, the device thickness, and relations of the discs to the atrial and aortic walls over time were evaluated by transesophageal echocardiography immediately and 3-12 months after deployment. RESULTS: The maximum unstretched ASD diameter was 16.2 +/- 4.8 mm and the device diameter selected was 20.6 +/- 5.5 mm. At the time of last follow-up, the device thickness decreased by 17-33%, 6 of 26 devices with a flare shape on the aortic side developed a closed shape, and the relations of the discs to the anterior atrial and aortic walls changed from touching to intermittent compression in 14 of the 78 cases. In these 14 cases, the aortic rim was significantly smaller, the number of flared device shapes on the aortic side/the number of closed shapes immediately after deployment was significantly larger, and the maximum device thickness at the middle part was significantly more decreased than those in other cases. CONCLUSION: As the device becomes thinner, loses its flexibility, and often changes from a flare-to-closed shape on the aortic side over time, the edges of ASO can start to compress the atrial and aortic walls. However, erosion was not recognized in these cases. PMID- 19281524 TI - Death of bamboo triggers regeneration of overstory tree in a southern beech forest. PMID- 19281525 TI - Kissing cousins: mycorrhizal fungi get together. PMID- 19281526 TI - Crassulacean acid metabolism under global climate change. PMID- 19281527 TI - Transport of TMV movement protein particles associated with the targeting of RNA to plasmodesmata. AB - The cell-to-cell movement of Tobacco mosaic virus through plasmodesmata (PD) requires virus-encoded movement protein (MP). The MP targets PD through the endoplasmic reticulum (ER)/actin network, whereas the intercellular movement of the viral RNA genome has been correlated with the association of the MP with mobile, microtubule-proximal particles in cells at the leading front of infection as well as the accumulation of the protein on the microtubule network during later infection stages. To understand how the associations of MP with ER and microtubules are functionally connected, we applied multiple marker three dimensional confocal and time-lapse video microscopies to Nicotiana benthamiana cells expressing fluorescent MP, fluorescent RNA and fluorescent cellular markers. We report the reconstitution of MP-dependent RNA transport to PD in a transient assay. We show that transiently expressed MP occurs in association with small particles as observed during infection. The same MP accumulates in PD and mediates the transport of its messenger RNA transcript to the pore. In the cellular cortex, the particles occur at microtubule-proximal sites and can undergo ER-associated and latrunculin-sensitive movements between such sites. These and other observations suggest that the microtubule network performs anchorage and release functions for controlling the assembly and intracellular movement of MP-containing RNA transport particles in association with the ER. PMID- 19281529 TI - Effect of Vibrio cholerae on chemokine gene expression in HT29 cells and its modulation by Lactobacillus GG. AB - Epithelial cells participate in the innate immune response to pathogenic bacteria by elaborating chemokines. This study examined the effect of Vibrio cholerae and Lactobacillus rhamnosus GG on inflammatory chemokine gene expression in the HT29 human intestinal epithelial cell line. HT29 cells were exposed to V. cholerae 0139, Lactobacillus or both for 2 h and cultured further thereafter for 4 h. RNA was extracted from the cells and expression of genes for chemokines and related molecules was quantitated by real time PCR using a pathway-focused PCR array. TLR4 was silenced using shRNA and output of interleukin-8 (IL-8) into the media quantitated with and without V. cholerae exposure. NFkappaB and p38 MAP kinase activation were determined by immunoblotting for IkappaBalpha and phosphorylated p38. Vibrio cholerae significantly upregulated gene expression for the neutrophil chemoattractant CXCL chemokines, IL-8, CXCL and CXCL in HT29 cells, while downregulating the expression of macrophage-attracting C-C chemokines. TLR4 silencing did not reduce IL-8 output from HT29 cells in response to V. cholerae. IkappaBalpha degradation was noted in the HT29 cells soon after exposure to V. cholerae and this recovered over time after removal of bacteria. p38 MAP kinase activation was not noted. Vibrio cholerae upregulated the expression of neutrophil attractant chemokines, most prominently IL-8, in HT29 cells, but downregulated macrophage-attracting chemokines. Probiotic lactobacilli modulated the IL-8, but not the other chemokine gene changes, in response to V. cholerae. PMID- 19281530 TI - Highly arthritis-susceptible DA rats express IL-1beta in articular cartilage. AB - There are genetically determined differences in susceptibility to arthritis among inbred rat strains. The aim of the present study was to elucidate phenotypical differences, by determining expression of TNF and IL-1beta, two pivotal mediators of arthritis, in the highly arthritis-prone Dark Agouti (DA) rat compared to that of two arthritis-resistant rat strains, the major histocompatibility complex homologous Piebald-Viral-Glaxo (PVG.1AV1) rat and the Brown Norway (BN) rat, assessed by immunohistochemistry. We demonstrate a distinct difference in articular cartilage, with chondrocytes expressing IL-1beta, not TNF, in the highly arthritis-prone DA rat as opposed to the two arthritis-resistant BN or PVG.1AV1 rat strains, where no cytokine expression was documented. The results were otherwise congruent among the rat strains. We observed TNF- and IL-1beta expressing cells within the synovial lining layer in all rat strains. Other tissues studied, auricular cartilage as well as muscle, lung, thyroid gland and kidney tissue, were devoid of cytokine expression. Constitutional expression of IL-1beta in chondrocytes might facilitate initiation and perpetuation of inflammation. This may offer one explanation of why erosive arthritides are so easily induced in the DA rat and also support the hypothesis that articular chondrocytes may themselves play a major role in cartilage matrix degradation. PMID- 19281531 TI - IC31, a two-component novel adjuvant mixed with a conjugate vaccine enhances protective immunity against pneumococcal disease in neonatal mice. AB - IC31 is a novel adjuvant which combines the immunostimulatory effects of an 11 mer antibacterial peptide (KLKL(5)KLK) and a synthetic oligodeoxynucleotide (ODN1a) which is a Toll-like receptor 9 agonist without containing cytosine phosphate guanine (CpG) motifs. The effects of IC31 on neonatal immune response to vaccination have not been reported. Neonatal mice were immunized once or twice with a Streptococcus pneumoniae serotype 1 polysaccharide conjugate containing Tetanus Toxoid (Pnc1-TT) carrier protein, with or without IC31 or CpG-ODN. IC31 significantly enhanced IgG1, IgG2a and IgG2b antibodies (Ab) to the serotype 1 polysaccharide. One dose of Pnc1-TT and low dose IC31 elicited high Ab levels that protected the neonatal mice completely from bacteraemia and significantly reduced lung infection following i.n. challenge with serotype 1 pneumococcal strain. One-sixth of an adult murine dose of IC31 was sufficient and optimal for induction of protective immunity in neonatal mice. Two doses of Pnc1-TT with or without adjuvants protected the neonatal mice completely, but more rapid Ab response was observed when IC31 was given with the Pnc1-TT. IC31 is a promising new adjuvant for neonatal vaccinations, rapidly enhancing protective humoral responses when combined with Pnc1-TT. PMID- 19281532 TI - Modified human beta 2-microglobulin (desLys(58)) displays decreased affinity for the heavy chain of MHC class I and induces nitric oxide production and apoptosis. AB - Beta2-microglobulin (beta2m) is the light chain of major histocompatibility complex class I (MHC-I) molecules, and is a prerequisite for the binding of peptides to the heavy chain and their presentation to CD8+ T cells. beta2m can be modified in vivo and in vitro by proteolytic cleavage by complement C1 and subsequent carboxypeptidase B-like activity--processes that lead to the generation of desLys(58) beta2m (dbeta2m). This work aims to study the effect of dbeta2m on peptide binding to MHC-I, the influence of dbeta2m on the binding of beta2m to the MHC-I heavy chain and the biological activity of dbeta2m. Both beta2m and dbeta2m are able to support the generation of MHC-I/peptide complexes at 18 degrees C, but complexes formed in the presence of dbeta2m destabilize at 37 degrees C. Moreover, a 250 times higher concentration of dbeta2m than of beta2m is needed to displace MHC-I associated beta2m from the cell surface. In addition, only beta2m is able to restore MHC-I/peptide complex formation on acid treated cells whereas dbeta2m appears to bind preferentially to denatured MHC-I heavy chains. In cell cultures, exogenously added dbeta2m, but not beta2m, induces apoptotic cell death in monocytic leukaemic cell lines but spares other kinds of leukaemic cells. Additionally, the presence of dbeta2m, and to a lesser extent beta2m, enhances IFN-gamma-induced NO production by monocytic leukaemic cells. In conclusion, these data show that dbeta2m is not able to support the formation of a stable tri-molecular MHC-I complex at physiological temperature and that dbeta2m exerts other biological functions compared to beta2m when bound to cells. PMID- 19281533 TI - Th1 cell reactivity and HLA-DR binding prediction for promiscuous recognition of MPT63 (Rv1926c), a major secreted protein of Mycobacterium tuberculosis. AB - MPT63 (Rv1926c), a major secreted protein of Mycobacterium tuberculosis, is immunoreactive in antibody assays in humans and animals and provides protection as a combined DNA vaccine in mice. This study was undertaken to determine the reactivity of MPT63 in T helper 1 (Th1) cell assays, i.e. antigen-induced proliferation and interferon-gamma secretion, using peripheral blood mononuclear cells (PBMCs) obtained from 72 Mycobacterium bovis Bacille Calmette-Guerin vaccinated healthy subjects. PBMCs were tested with complex mycobacterial antigens and pools of synthetic peptides corresponding to MPT63, MPB70, MT24, PPE68, CFP10 and ESAT-6. The results showed that MPT63 induced moderate Th1 cell reactivity which was equivalent to the reactivity induced by other secreted antigens of M. tuberculosis, i.e. MT24 and MPB70. Furthermore, human leucocyte antigen (HLA) heterogeneity of the responding donors suggested that MPT63 was presented to Th1 cells promiscuously. Analysis of the MPT63 sequence and its peptides for binding to 51 alleles of 9 serologically defined HLA-DR molecules, using a virtual matrix-based prediction program (ProPred) showed that MPT63 sequence could bind to all the 51 alleles, whereas 9 of the 10 peptides of MPT63 were also predicted to bind promiscuously. When tested with PBMCs of HLA-DR heterogeneous donors that responded to MPT63 in interferon-gamma assays, at least 9 of the 10 peptides of MPT63 were recognized by PBMCs from HLA heterogeneous donors. These results suggested that promiscuous Th1 cell reactive epitopes are scattered throughout the sequence of MPT63, and further support the inclusion of this protein in an antigen cocktail to develop a new anti-tuberculosis vaccine. PMID- 19281534 TI - Effects of human fecal flora on intestinal morphology and mucosal immunity in human flora-associated piglet. AB - Human flora-associated (HFA) piglet model was established to examine the effects of gut microbes from a different donor species on the intestinal morphology and mucosal immunity. Newborn germ-free piglets, obtained by caesarean section, were orally inoculated with a human and a porcine faecal suspension, and artificially fed to establish a HFA group (n = 7) and pig flora-associated (PFA) group (n = 7), respectively. All pigs were killed 6 weeks later. Tissue samples from duodenum, jejunum, ileum and colon were collected and studied by histochemistry and immunohistochemistry methods for intestinal morphological analyses and detection of immunocompetent cells. In summary, both groups of pigs performed well but HFA pigs had a somewhat better daily weight gain, and their jejunal villus height and crypt depth were significantly higher. In comparison with PFA pigs, the number of intraepithelial lymphocytes in jejunum was lower but the number of goblet cells containing neutral mucins was significantly increased in HFA pigs. No difference was observed in the number of mast cells. The areas of IgA producing cells and CD4(+) T cells in the jejunum and IgG producing cells in the small intestine were significantly higher in HFA pigs. However, the areas of MHC class II expressing cells were significantly increased in the duodenum and colon. Additionally, the amount of Bifidobacteria spp. was significantly higher in HFA pigs. This study confirms that the composition of gut microbes differentially affects the host intestinal mucosal immunity and suggests that commensal bacteria have great effects on intestinal health and development. PMID- 19281535 TI - Effects of haemofiltration and mannitol treatment on cardiopulmonary-bypass induced immunosuppression. AB - Cardiac surgery using cardiopulmonary bypass (CPB) causes a systemic inflammatory response. Additionally, an impairment of the responsiveness of peripheral blood mononuclear cells (PBMC) to further immunological stimuli has been observed. The aim of our present study was to evaluate the ability of antioxidant therapy with mannitol or haemofiltration during CPB to modulate this immunosuppression after CPB. Forty-five patients undergoing elective heart-surgery were prospectively enrolled and randomized into three groups (control, mannitol, haemofiltration). Blood samples were taken after induction of anaesthesia (T1), 20 min after CPB (T2) and 24 h post-operatively (T3). Expression density of the monocytic surface receptor CD14, HLA-DR expression and cytokine release (TNF-alpha and IL10) after lipopolysaccharide-stimulation were evaluated. At T2, the CD14(dim) cell population was maintained in both intervention groups while in the control group there was a decrease of this proinflammatory monocytic phenotype. No significant differences regarding HLA-DR expression or cytokine release could be demonstrated. This study shows that the suppression of the stimulated immune response after CPB can potentially be alleviated by mannitol or haemofiltration in an experimental in-vitro setting. In the light of data showing that this depression of the immune response might affect the post-operative course of patients, these results could have a potential clinical relevance. PMID- 19281536 TI - Effect of an extract based on the medicinal mushroom Agaricus blazei murill on release of cytokines, chemokines and leukocyte growth factors in human blood ex vivo and in vivo. AB - An immunostimulatory extract based on the medicinal mushroom Agaricus blazei Murill (AbM) has been shown to stimulate mononuclear phagocytes in vitro to produce pro-inflammatory cytokines, and to protect against lethal peritonitis in mice. The present aim was to study the effect of AbM on release of several cytokines in human whole blood both after stimulation ex vivo and in vivo after oral intake over several days in healthy volunteers. The 17 signal substances examined were; T helper 1 (Th1) cytokines [interleukin (IL)-2, interferon (IFN) gamma and IL-12], T helper 2 cytokines (IL-4, IL-5 and IL-13), pleiotropic (IL-7, IL-17), pro-inflammatory [IL-1beta, IL-6, tumour necrosis factor (TNF)-alpha (mainly produced by Th1 cells)]--and anti-inflammatory (IL-10) cytokines, chemokines [IL-8, macrophage inhibitory protein (MIP)-1beta and monocyte chemoattractant protein (MCP)-1] and leukocyte growth factors [granulocyte colony stimulating factor (G-CSF), granulocyte/macrophage colony stimulating factor]. After stimulation of whole blood ex vivo with 0.5-5.0% of a mushroom extract, AndoSan mainly containing AbM, there was a dose-dependent increase in all the cytokines studied, ranging from two to 399-fold (TNF-alpha). However, in vivo in the eight volunteers who completed the daily intake (60 ml) of this AbM extract for 12 days, a significant reduction was observed in levels of IL-1beta (97%), TNF-alpha (84%), IL-17 (50%) and IL-2 (46%). Although not significant, there was a trend towards reduced levels for IL-8, IFN-gamma and G-CSF, whilst those of the remaining nine cytokines tested, were unaltered. The discrepant results on cytokine release ex vivo and in vivo may partly be explained by the antioxidant activity of AbM in vivo and limited absorption of its large, complex and bioactive beta-glucans across the intestinal mucosa to the reticuloendothelial system and blood. PMID- 19281537 TI - An alkaloid extract of Evanta, traditionally used as anti-Leishmania agent in Bolivia, inhibits cellular proliferation and interferon-gamma production in polyclonally activated cells. AB - Traditional medicine and scientific studies have shown that the raw extract of Evanta [Galipea longiflora, Angostura longiflora (Krause) Kallunki] exhibits anti leishmanial activity. We hypothesized that the healing observed when using this plant might not only be due to the direct action on the parasite, but possibly to a parallel effect on the host immune response to the parasite involved in the healing process. We show here that an alkaloid extract of Evanta (AEE) directly killed the parasite already at a dose of 10 microg/ml, but at this low concentration, AEE did not have a major effect on viability and proliferation of eukaryotic cells. The whole extract was also found to be stronger than 2 phenylquinoline, the most prominent alkaloid in AEE. AEE was not directly stimulating B or T cells or J774 macrophages. However, it interfered with the activation of both mouse and human T cells, as revealed by a reduction of in vitro cellular proliferation and interferon-gamma (IFN-gamma) production. The effect was more evident when the cells were pretreated with AEE and subsequently stimulated with the polyclonal T-cell activators Concanavalin A and anti-CD3. Taken together, our results suggest that Evanta have a direct leishmanicidal effect and due to the effect on IFN-gamma production it might contribute to control the chronic inflammatory reaction that characterize Leishmania infection pathology, but in vivo studies are necessary to corroborate this finding. PMID- 19281538 TI - Relationship between circulating levels of IFN-gamma, IL-10, CXCL9 and CCL2 in pulmonary and extrapulmonary tuberculosis is dependent on disease severity. AB - Protection against Mycobacterium tuberculosis infection is dependent on T cell and macrophage activation regulated by cytokines. Cytokines and chemokines produced at disease sites may be released into circulation. Data available on circulating cytokines in tuberculosis (TB) is mostly on pulmonary TB (PTB) with limited information on extrapulmonary disease (EPul-TB). We measured interferon gamma (IFN-gamma), interkeukin-10 (IL-10), CXCL9 and CCL2 in sera of patients (n = 80) including; PTB (n = 42), EPul-TB (n = 38) and BCG vaccinated healthy endemic controls (EC, n = 42). EPul-TB patients comprised those with less severe (LNTB) or severe (SevTB) disease. Serum IFN-gamma, IL-10 and CXCL9 levels were significantly greater while CCL2 was reduced in TB patients as compared with EC. IFN-gamma was significantly greater in PTB as compared with LNTB (P = 0.002) and SevTB (P = 0.029). CXCL9 was greater in PTB as compared with LNTB (P = 0.009). In contrast, CCL2 levels were reduced in PTB as compared with LNTB (P = 0.021) and SevTB (P = 0.024). A Spearman's rank correlation analysis determined a positive association between IFN-gamma and IL-10 (rho = 0.473, P = 0.002) and IFN-gamma and CXCL9 (rho = 0.403, P = 0.008) in the PTB group. However, in SevTB, only IFN gamma and CXCL9 were positively associated (rho = 0.529, P = 0.016). Systemic levels of cytokines are reflective of local responses at disease sites. Therefore, our data suggests that in PTB increased IFN-gamma and CXCL9 balanced by IL-10 may result in a more effective cell mediated response in the host. However, elevated inflammatory chemokines CXCL9 and CCL2 in severe EPul-TB without concomitant down modulatory cytokines may exacerbate disease related pathology and hamper restriction of M. tuberculosis infection. PMID- 19281539 TI - Bronchoalveolar lavage fluid cellular characteristics, functional parameters and cytokine and chemokine levels in interstitial lung diseases. AB - Idiopathic pulmonary fibrosis (IPF), hypersensitivity pneumonitis (HP) and sarcoidosis belong to interstitial lung diseases (ILD) where an imbalance of regulatory, profibrotic and antifibrotic cytokines is hypothesized. The relationship of bronchoalveolar lavage (BAL) fluid (BALF) cytokines, BALF cell profile and ILD course is supposed. The aim of our study was to correlate BALF cytokine and chemokine levels with BALF cellular characteristics and lung function parameters in different ILD. Twenty-two sarcoidosis, seven IPF and 11 HP patients underwent lung function tests and BAL. The BALF differential cell counts and superficial cell markers were characterized, and MCP-1, MIP-1alpha, MIP 1beta, RANTES, epithelial neutrophil-activating protein (ENA)-78, FGF, G-CSF, GM CSF, IFN-gamma, interleukin (IL)-1alpha, IL-1RA, IL-1beta, -2beta, -4beta, 5beta, -6beta, -8beta, -10beta, -17beta, tumour necrosis factor (TNF)-alpha, thromobopoietin (Tpo) and vascular endothelial growth factor (VEGF) values measured. The BALF VEGF values were highest in sarcoidosis (P = 0.0526). IL-1RA values were higher in IPF and HP compared with sarcoidosis (P = 0.0334). IL-8/ENA 78 ratio positively correlated with BALF neutrophil counts in IPF (r = 0.89, P = 0.04). Vital capacity and TL(CO) values positively correlated with VEGF and negatively with IL-8 BALF levels in all ILDs but the correlations were most significant in sarcoidosis group. We suppose that VEGF plays a role in ILDs' early phases and has rather angiogenic than profibrotic effect. On the contrary, IL-8 is probably upregulated in advanced ILDs with prominent fibrosis and marked lung functions decline. We state that BALF VEGF, IL-8 and ENA-78 levels and IL 8/ENA-78 ratio could become useful markers of ILDs' phase, activity and prognosis. They might also be helpful in treatment modality choice. PMID- 19281540 TI - Natural immunity has significant impact on immune responses against cancer. AB - The immune system defends the host against pathogenic attacks by micro-organisms and their products. It does not react against self-components due to the relatively efficient negative selection of developing T lymphocytes in the thymus. This process does permit T cells with low avidity against self to be present in the T cell repertoire. Such cells play an important physiological role as the host needs so-called autoimmune reactions in order to eliminate dying cells or transformed tumour cells. One of the mysteries in immunology is how the host maintains beneficial autoimmune reactions and avoids pathogenic autoimmune reactions. Activation of the adaptive T lymphocytes is mediated by the low avidity interaction between T-cell antigen receptors and antigenic peptides associated with major histocompatibility complex class I or class II molecules. This interaction is strengthened by T-cell co-receptors such as CD2, CD4, CD8, CD28 and CD154, which react with ligands expressed by cells of the innate immune system. In recent years, the importance of pre-activation of the innate immune system for initiation of adaptive T-cell immune responses has been appreciated. In the present review, I will summarize our work on how natural immunity plays an important role in determining the level of beneficial autoimmune reactions against cancer. PMID- 19281541 TI - Associations between ward atmosphere, patient satisfaction and outcome. AB - Previous studies have found important associations between the ward atmosphere and patient satisfaction. However, fewer studies have examined the relationship between ward atmosphere and outcome of treatment. The aim of the study was to examine whether or not differences in ward atmosphere were associated with differences in satisfaction and outcome. Eighty patients at three different ward units responded to a questionnaire at admission and by the time of discharge from the hospital. The questionnaire comprised the ward atmosphere scale, a five-item index of patient satisfaction, the Generalized Self-Efficacy Scale, an index of life satisfaction and the symptom checklist SCL-90R. The results showed that differences in the treatment environment between the ward units were associated with differences in patient satisfaction. There was mixed evidence for associations between ward atmosphere and outcome, while no associations were found between ward atmosphere and self-efficacy and life satisfaction. The results may suggest that the relationship between ward atmosphere and outcomes of treatment may be of a more indirect character than the relationship between ward atmosphere and satisfaction. PMID- 19281542 TI - Use of a think-aloud procedure to explore the relationship between clinical reasoning and solution-focused training in self-harm for emergency nurses. AB - Self-harm is a risk factor for further episodes of self-harm and suicide. The most common service used by self-injurers is the emergency department. However, very often, nurses have received no special training to identify and address the needs of these patients. In addition this care context is typically biomedical and without psychosocial skills, nurses can tend to feel unprepared and lacking in confidence, particularly on the issue of self-harm. In a study that aimed to improve understanding and teach solution-focused skills to emergency nurses so that they may be more helpful with patients who self-harm, several outcome measures were considered, including knowledge, professional identity and clinical reasoning. The think-aloud procedure was used as a way of exploring and improving the solution-focused nature of nurses' clinical reasoning in a range of self-harm scenarios. A total of 28 emergency nurses completed the activity. Data were audiotaped, transcribed and analysed. The results indicated that significant improvements were noted in nurses' ability to consider the patients' psychosocial needs following the intervention. Thus this study has shown that interactive education not only improves attitude and confidence but enlarges nurses' reasoning skills to include psychosocial needs. This is likely to improve the quality of care provided to patients with mental health problems who present to emergency settings, reducing stigma for patients and providing the important first steps to enduring change - acknowledgment and respect. PMID- 19281543 TI - Evaluation of a community-based mental health drug and alcohol nurse in the care of people living with HIV/AIDS. AB - There is a growing need for advanced practice mental health and drug and alcohol nursing roles in the care of people living with HIV/AIDS; however, limited publications address these domains. This study evaluated a community-based mental health drug and alcohol nurse role caring for people living with HIV/AIDS (Mental Health D&A Nurse) in a large not-for-profit district nursing organization providing care to people living with HIV/AIDS in an Australian city. Outcomes from a client assessment and 6-8-week follow-up by the Mental Health D&A Nurse are presented as captured by the Depression Anxiety Stress Scales (DASS 21), Alcohol, Smoking and Substance Involvement Screening Test (ASSIST), Health of the Nation Outcome Scales (HONOS) and WHOQoL BREF. Mean scores and caseness were analysed, and significant differences were found on the 'impairment' and 'social problems' subscales of the HONOS. Results of semi-structured interviews with clients describe effective and supportive mental health care and health-promoting education following visits by the Mental Health D&A Nurse. These positive findings support continuing implementation of the role within this community setting and indicate that even greater benefits will ensue as the role develops further. Findings are of interest to clinicians and policy makers seeking to implement similar roles in community-based HIV/AIDS care. PMID- 19281544 TI - Resilience revisited. AB - Contemporary reports of individuals experiencing severe adversity have led to renewed debate regarding the processes that mediate a person's ability to withstand extremes of both psychological and physical abuse. The capacity to recover from extremes of trauma and stress is termed resilience. This important concept is of key relevance to health professionals as resilience and also defines a more general concept of responding to challenges that affect the individual in terms of their health behaviour. The evidence base in this area is noteworthy in that much research is based on historical case studies that, although rich in both content and appeal, also fall short of contemporary rigour in terms of validity. This review will describe the evidence for the salience of the concept of the resilience to health and mental health and the implications of the domain to the healthcare professional. PMID- 19281545 TI - The competence of certified nurse assistants caring for persons with dementia diseases in residential facilities. AB - In Sweden, a majority of elderly persons living in residential facilities have some form of dementia, and caring for them is a demanding and complex task. The formal education level of personnel in residential facilities is low. The aim of this paper is to describe certified nurse assistants' (CNAs) content of competence caring for persons with dementia diseases in residential facilities. Information from 22 diary notes was obtained and 12 CNAs in four residential facilities were interviewed. The diary notes and interviews were transcribed and analysed using conventional content analysis. Three main categories were generated from the diaries: Social and Health Care, Health Care Targeted towards Medicine and, finally, Organization. The same categories were generated from the interviews but with the addition of Knowledge - its use, growth and deficiencies. The results show that competence is mainly based on personal, experienced-based knowledge and 'standard routines'. In general, the nurse assistants' interest in developing knowledge is weak. Furthermore, CNAs lack competence in essential theoretical knowledge about dementia diseases. It is notable that CNAs, caring for people with serious dementia, know little about the reasons for the residents' divergent behaviour. Consequently, knowledge related to dementia and mental diseases must be highlighted. Therefore, formal education focused on these diseases is crucial. PMID- 19281546 TI - Epidemiology of absconding from an Iranian psychiatric centre. AB - Absconding by psychiatric patients from acute psychiatric wards has been a significant problem for some time. This study was conducted to identify the incidence rate of absconding in a psychiatric hospital and some of the characteristics of patients who had escaped from hospital in year 2006. In this descriptive cross-sectional study, all of absconds in our centre have been registered in the period of 9 months. We assessed patients' age, gender, marital status, hospital stay, substance abuse, psychiatric diagnosis and site of absconding. A total of 43 cases of absconding have been registered in this period, attempted by 35 patients, with the incidence rate of 3% in our centre. The mean age of absconders was 31.7 years; 95.3% of absconds were made by men; 67.4% were single; 37.4% absconded through the main door; 62.8% of our absconders had bipolar mood disorder or schizophrenia; mean in hospital stay before absconding was 18.4 days; and 53.5% of absconds were attempted by substance users. Absconders utilize a significant portion of hospital resources and have an adverse effect on ward milieu. It seems that the patients who are more prone to abscond are male, young, single, addict, with the diagnosis of bipolar mood disorder or schizophrenia and patients who are admitted not long ago. PMID- 19281547 TI - Binary constructs of forensic psychiatric nursing: a pilot study. AB - The aim was to develop an Information Gathering Schedule (IGS) relevant to forensic psychiatric nursing in order to establish the perceived differences in the three levels of security, high, medium and low. Perceived differences in the role constructs of forensic psychiatric nursing is said to exist but the evidence is qualitative or anecdotal. This paper sets out a pilot study beginning in 2004 relating to the development of two rating scales for inclusion into an IGS to acquire data on the role constructs of nurses working in these environments. Following a thematic analysis from the literature two sets of binary frameworks were constructed and a number of questions/statements relating to them were tested. The Thurstone Scaling test was applied to compute medians resulting in a reduction to 48 and 20 items for each respective framework. Two 7-point Likert scales were constructed and test-retest procedures were applied on a sample population of forensic psychiatric nurses. Student's t-test was conducted on the data and the results suggest that the IGS is now suitable for application on a larger study. The IGS was piloted on a small sample of forensic psychiatric nurses. The two scales were validated to coefficient values ranging from 0.7 to 0.9. Amendments were made and the IGS was considered acceptable. PMID- 19281548 TI - An exploration of the social identity of mental health inpatient service users. AB - Social identity theory proposes that a threatened social identity can impact on self-concept and well-being. As a low-status minority group, mental health service users face the possibility of a threatened social identity compounding existing mental health problems. This may be further complicated by an inpatient admission where the context in which social identity is maintained has changed from a community setting. We use a qualitative approach to explore the salient social identity of mental health inpatient service users using semi-structured interviews. Analysis showed that the salient in-groups and out-groups mainly centred around the inpatient context but were somewhat influenced by non-mental health service group membership. Participants also talked of their unique characteristics as group members. How these findings relate to social identity theory and, particularly how they relate to mental health service users within changing contexts is explored. PMID- 19281549 TI - Community treatment orders: the ethical balancing act in community mental health. AB - Community treatment orders (CTOs) are legal mechanisms by which an individual with a mental illness and a history of non-compliance and potential for violence can be mandated (against their will) to undergo psychiatric treatment in an outpatient setting. Although CTOs are increasingly being adopted by governments as a means of protecting both mentally ill persons and society itself, their use continues to stimulate considerable debate. While there is some evidence of their potential benefits in promoting treatment compliance and reducing hospital stays, there is concern that they infringe on the mental health client's human rights and freedoms. Consideration of the ethical and practical implications of the use of CTOs must continue. In this paper, some of the most pressing issues are identified and discussed. PMID- 19281550 TI - Advocacy in mental health nursing: an integrative review of the literature. AB - The term 'advocacy' has taken on a meaning beyond its legal origins and is now of importance as a concept in health and social care. Within nursing, the role of advocate has been accepted as an important one, although there are arguments against nurses taking on such a role. The majority of papers on this topic relate to nursing generically rather than being specifically mental health oriented. Nurses need to be made aware of the legal framework within which they practice, in terms of duty of care within their role of nurse advocate, maintaining standards of advocacy acceptable to their professional body, accountability relating to action and omission of actions, guidance on guarding against stepping beyond the boundaries of their professional practice of advocacy, and to have adequate knowledge of the law. This paper offers a critique of nursing advocacy models, raising a number of mental health nursing issues and identifying some areas for further research. PMID- 19281551 TI - Burnard (2007): autoethnography or a realist account? PMID- 19281552 TI - Misplaced epistemological certainty and pharmaco-centrism in mental health nursing. PMID- 19281553 TI - Why cause matters. AB - Recently, some commentators have suggested, for either pragmatic or philosophical reasons, that the cause of mental distress is irrelevant. In this paper it is suggested that downplaying the role of cause leads to ignoring the link between abuse and mental health difficulties which is disempowering for individuals, short-sighted for service provision and fails to challenge the social structures, inequalities, conditions, practices, values and attitudes that allow abuse to happen. Acknowledging the causal link between abuse and mental health difficulties implies that in addition to building recovery focused therapeutic relationships with individuals, as mental health nurses we need to take seriously our role in mental health promotion from infancy onwards and to acknowledge the importance of being politically aware and involved. PMID- 19281554 TI - A retrospective analysis of absconding behaviours by psychiatric inpatients in one psychiatric hospital campus in Australia. PMID- 19281563 TI - Divergence without difference: phylogenetics and taxonomy of Chlamydia resolved. AB - Members of Chlamydiaceae have been extensively characterized by complete genome sequencing. This information provides new understanding concerning their natural evolutionary history. Comparative genome analysis is consistent with the conclusion that host-divergent strains of Chlamydiae are closely related biologically and ecologically. The previous taxonomic separation of the genus based on ribosomal sequences is neither consistent with the natural history of the organism revealed by genome comparisons, nor widely used by the Chlamydia research community 8 years after its introduction; thus, it is proposed to reunite the Chlamydiaceae into a single genus, Chlamydia. PMID- 19281564 TI - Typing Chlamydia trachomatis: from egg yolk to nanotechnology. AB - A historical review is provided of the various methods used for half a century to differentiate and type Chlamydia trachomatis strains. Typing of C. trachomatis is an important tool for revealing transmission patterns in sexual networks, and enabling association with clinical manifestations and pathogenicity. Serotyping using the major outer membrane protein (MOMP) has been the mainstay of epidemiological work for several decades. However, the development of nucleic acid amplification techniques (NAAT) and easy access to sequencing have shifted the focus from MOMP serotypes to omp1 genotypes. However, insufficient epidemiological resolution is achieved by characterization of both MOMP and omp1. This calls for new high-resolution genotyping methods applying for example a multilocus variable number tandem repeat assay (MLVA) or multilocus sequence typing (MLST). The futuristic nanotechnology already seems at hand to further simplify and automate the high-resolution genotyping method based on NAAT and sequencing of various targets in the C. trachomatis genome. Thereby, a high throughput can be achieved and more epidemiological information can be obtained. However, it is important to realize that culture of C. trachomatis may still be needed to detect and characterize new variants of C. trachomatis. PMID- 19281565 TI - Chlamydia pneumoniae-induced pathological signaling in the vasculature. AB - Since its description in 1986, Chlamydia pneumoniae has remained one of the most enigmatic pathogens. This intracellular bacterium is highly seroprevalent, but rarely recovered from cell culture, it can genetically switch between a proliferative and a nonreplicative state and has been linked to a vast number of chronic diseases, most notably to atherosclerosis, as it can be found in the plaques. It has become quite clear that persistent bacteria in atherosclerotic lesions cannot be eradicated by currently available antibiotic treatments and that attempts to do so without a better understanding of the pathobiology of chlamydial persistence are futile. However, there is growing knowledge on how vascular chlamydial infection may lead to the pathological reprogramming of the host cell signaling pathways. Chlamydia pneumoniae is now well known to induce, at least in vitro, the two pathogenetic main events that define atherosclerosis: angiogenesis and inflammation. In vivo a contribution of chlamydial infection to the progression of atherosclerosis remains unproven. This minireview provides a brief overview on the proproliferative and proinflammatory effects of vascular C. pneumoniae infection and their potential link to atherogenesis. PMID- 19281566 TI - Apoptosis resistance in Chlamydia-infected cells: a fate worse than death? AB - Chlamydia has long been studied as an intracellular pathogen causing widespread diseases. In the last three decades, the field of apoptosis has rapidly emerged, and as a consequence, research on infectious diseases in general and on Chlamydia host interaction in particular shifted to apoptosis modulation. Ten years ago, the first paper describing the drastic inhibition of apoptosis in Chlamydia infected cells was published. In a reversal of roles, here was a pathogen that was strongly protecting cells in an organism against destruction by the organism's immune system. Since then, numerous studies have described apoptosis inhibition by Chlamydia and the mechanisms involved, but still there is a lack of general consensus on the subject. With a section of studies even reporting the induction of cell death by Chlamydia and not its inhibition, the field became even more diverse and complicated. In this review, an attempt is made to discuss the recent findings on apoptosis modulation by chlamydial species. PMID- 19281567 TI - In vitro neutralization of tumor necrosis factor-alpha during Chlamydia pneumoniae infection impairs dendritic cells maturation/function and increases chlamydial progeny. AB - Dendritic cells (DCs) produce tumor necrosis factor (TNF)-alpha upon infection and contribute in various ways to defense against pathogenic agents. Several biological agents have been designed to inhibit TNF-alpha activity. However, the use of these inhibitors has been associated with an increased rate of certain opportunistic infections. To study the effect of TNF-alpha inhibition, human monocyte-derived DCs were infected with Chlamydia pneumoniae. TNF-alpha was neutralized by adalimumab, a human anti-TNF-alpha monoclonal antibody. Chlamydiae induced the maturation of DC as determined by flow cytometry and quantitative real-time PCR. However, DC maturation was impaired in the presence of adalimumab. Moreover, neutralization of TNF-alpha resulted in a significant increase of infectious progeny, 16S rRNA gene copy number and development of larger inclusions consisting of different stages of chlamydial development. Additionally, chlamydial infection induced secretion of cytokines/chemokines, which were downregulated by adalimumab treatment. Our data reveal an indirect effect on maturation of DC by C. pneumoniae and that maturation is crucial for the restriction of chlamydial development. The results also demonstrate an increase in infectious progeny after TNF-alpha inhibition, suggesting a contribution of TNF-alpha produced by DCs to chlamydial growth arrest. These data suggest a possible mechanism by which TNF-alpha inhibition enhances the risk of intracellular infections. PMID- 19281568 TI - Identification and characterization of novel recombinant vaccine antigens for immunization against genital Chlamydia trachomatis. AB - Chlamydia trachomatis infection is the most common sexually transmitted bacterial infection worldwide, with over 91 million cases estimated annually. An effective subunit vaccine against Chlamydia may require a multivalent subunit cocktail of antigens in a single formulation for broad coverage of a heterogeneous major histocompatibility complex population. Herein, we describe the identification of novel C. trachomatis antigens by CD4+ and CD8+ T-cell expression cloning, serological expression cloning, and an in silico analysis of the C. trachomatis genome. These antigens elicited human CD4+ T-cell responses, and a subset proved to be immunogenic and protective when administered as immunoprophylactic vaccines against C. trachomatis challenge. Candidate vaccines consisting of the prioritized C. trachomatis antigens adjuvanted in a GlaxoSmithKline proprietary AS01B adjuvant were prioritized based on induction of solid protection against challenge in C57BL/6 and BALB/c mice with C. trachomatis. Some of the vaccines prevented bacterial shedding and colonization of the upper genital tract to varying degrees by mechanisms that may include CD4+ T cells. PMID- 19281569 TI - A limited role for antibody in protective immunity induced by rCPAF and CpG vaccination against primary genital Chlamydia muridarum challenge. AB - Mice deficient in B cells (micromT mice) were used to evaluate the role of antibody in enhanced chlamydial clearance and reduction of pathology afforded by vaccination with recombinant chlamydial protease-like activity factor (rCPAF). Enhanced, but comparable, chlamydial clearance was observed in micromT and wild type (WT) mice after rCPAF+CpG vaccination. Chlamydia-induced pathology was present in mock-immunized animals, but at significantly greater levels in micromT than WT mice, whereas vaccinated micromT and WT mice exhibited similar reductions in pathology. Thus, antibodies may play a role in protection against chlamydial pathology after primary infection, but were largely dispensable in rCPAF+CpG induced chlamydial clearance and reduction in pathology. PMID- 19281570 TI - As good as it gets in fetal therapy? PMID- 19281571 TI - Hereditary angioedema and pregnancy. AB - BACKGROUND: Hereditary angioedema (HAE) is an autosomal dominant disease caused by a quantitative or functional defect in C1-esterase inhibitor (C1-INH). Patients with this deficiency present with episodes of angioedema which can be life-threatening. Studies examining HAE and pregnancy are scarce with little known about the interrelationship between the two. OBJECTIVE: To examine the effect, and evaluate the clinical manifestations of HAE in pregnancy using retrospective interviews of affected women. METHODS: Women with HAE who have undergone one of more pregnancies were identified throughout Australia using the national Australasian Society of Clinical Immunology and Allergy immunodeficiency database. Following informed consent, identified women were interviewed regarding their HAE status during pregnancy and the perinatal period using a questionnaire. RESULTS: Seven women with a total of 16 pregnancies were identified. During the first trimester of pregnancy, more than ten attacks of angioedema were experienced in six of 16 pregnancies. During the second trimester only in three of 16 pregnancies did women experience greater than ten attacks. During the post partum period, four of seven women experienced increased frequency and severity of attacks as compared to the pre-pregnancy state. For two of four patients, this impacted on their breast-feeding routine. CONCLUSION: Our study showed that women with HAE have greatly reduced or absent attacks in the last two trimesters of pregnancy, although, during the post-partum period, the majority of women experienced increased frequency and severity of attacks. PMID- 19281572 TI - An overview of the literature on congenital lower urinary tract obstruction and introduction to the PLUTO trial: percutaneous shunting in lower urinary tract obstruction. AB - Congenital lower urinary tract obstruction (LUTO) comprises a heterogeneous group of pathologies causing obstruction to the urethra, the most common being posterior urethral valves. Such pathology is often associated with high perinatal mortality and varying degrees of perinatal and infant morbidity. A high proportion of LUTO may be visualised during routine second trimester (and first trimester) ultrasound giving rise to the possibility of determining individual fetal prognosis and treatments such as vesico-amniotic shunting, with a view to altering pathogenesis. The aims of the percutaneous shunting in low urinary tract obstruction (PLUTO) trial are to determine the effectiveness of these treatments and accuracy of the investigations with the primary outcome measures being perinatal mortality and postnatal renal function. PMID- 19281573 TI - Association of maternal pre-pregnancy weight with birth defects: evidence from a case-control study in Western Australia. AB - BACKGROUND: Maternal obesity confers increased risks of poor pregnancy outcomes. There are limited Australian data on the risk of birth defects associated with maternal pre-pregnancy obesity. METHODS: Population-based case-control study of 418 controls, 111 cases with heart defects (and of these, 38 had conotruncal heart defects), 27 with neural tube defects, 86 cases with urinary tract defects, 48 cases with orofacial clefts, and 20 with limb reduction defects. Maternal pre pregnancy weight and height were self-reported. RESULTS: Women with pre-pregnancy obesity (body mass index 30+) had a twofold increased odds of having an infant with neural tube defects, conotruncal heart defects, orofacial clefts and limb reduction defects and 30-40% increase in heart defects generally and urinary tract defects. None of the estimates was statistically significant. CONCLUSIONS: Our findings were consistent with similar, statistically significant studies in the literature. Weight reduction prior to pregnancy in obese women may be a means of primary prevention of birth defects. PMID- 19281574 TI - Clinical predictive factors for preterm birth in women with threatened preterm labour or preterm premature ruptured membranes? AB - BACKGROUND: Independent predictive factors of preterm delivery were evaluated using clinical data at hospitalisation by multivariate analysis. AIM: The aim of this study was to clarify independent predictive factors related to preterm delivery by multivariate analysis of clinical data at hospitalisation of patients with threatened preterm delivery or premature rupture of membranes (PROM), and to realise the early and highly reliable prediction of preterm delivery in pregnant women at risk. METHODS: The subjects were 200 patients, which diagnosed with threatened preterm delivery or PROM and admitted at gestational ages of 22-35 weeks. Univariate and multivariate analyses were performed; 20 factors were evaluated concerning clinical data, and we extracted prognostic factors using logistic regression analysis. RESULTS: The mean age of the patients was 30.5 years, and the mean gestational age at admission was 30.0 weeks. Preterm delivery was observed in 55 (27.5%), and term delivery in 145 (72.5%). On multivariate analysis, haemorrhage, prepregnancy body mass index, fetal fibronectin and cervical length were extracted as independent predictive factors related to preterm delivery. CONCLUSIONS: If the reliable and reproducible prediction of preterm delivery becomes possible using the four factors extracted in this study, further evaluation of these factors may lead to clarification of the mechanism of preterm delivery. PMID- 19281575 TI - Selective fetoscopic laser ablation in 100 consecutive pregnancies with severe twin-twin transfusion syndrome. AB - AIMS: To report the perinatal outcomes of a large series of twin pregnancies with severe twin-twin transfusion syndrome (TTTS) managed with laser ablation surgery in an Australian tertiary perinatal centre and to compare the outcome with other large cohorts. METHODS: The outcomes of 100 consecutive pregnancies with severe TTTS managed with selective fetoscopic laser ablation from March 2002 to June 2007 were examined. Survival and neonatal morbidity were analysed. Comparisons were made with the results from other studies of laser surgery with at least 100 pregnancies. RESULTS: There were 100 women with TTTS treated with laser ablation; 34 stage II, 44 stage III and 22 at stage IV. Median gestation at time of laser was 21 weeks (range 18-28) and median gestation at delivery was 31 weeks (range 20-39). Overall perinatal survival rate was 151 of 200 (75.5%). Eighty five per cent had one or more surviving twins. The survival rate for stage IV TTTS was 88.6%, significantly better than for stage II (69.1%) and stage III (73.9%) pregnancies. The perinatal mortality rate for donors (30%) was not significantly different from recipients (19%), but the fetal death rate for donors was significantly greater than that for recipients (P = 0.03). Severe cerebral abnormalities were present in only 2.8% of newborns. The overall survival rate was comparable to other large series. CONCLUSIONS: These results for the management of severe TTTS are comparable to the best reported international series. Long-term follow-up is required and more research needs to be undertaken to further improve these results. PMID- 19281576 TI - The selective use of rapid aneuploidy screening in prenatal diagnosis. AB - AIMS: To evaluate the diagnostic utility and costing of the selective use of rapid aneuploidy screening (RAS) for chorion villus sampling (CVS) and amniocentesis specimens. METHODS: CVS and amniocenteses performed between 2000 and 2006 were identified. Cases were subdivided into two groups: (i) RAS in addition to long-term culture and (ii) long-term chromosome culture alone. The frequency of RAS, the proportion of abnormal results and the cytogenetic costings were reviewed. RESULTS: A total of 3315 procedures were performed: 730 CVS and 2585 amniocenteses. An abnormal karyotype culture was present in 366 of 3315 (11%). For CVS an abnormal culture was present in 164 (22.5%). RAS (short-term culture/direct preparation) was selectively used in 399 cases (54.6%) with an abnormal result in 128 (32% of RAS). For amniocentesis, 206 chromosome abnormalities were present (8.0% of specimens). RAS (interphase FISH) was selectively used in 580 amniocenteses (22.4%). FISH was requested in 95 (66.4%) of the 143 abnormal cases potentially detectable with standard probes. There was a progressive increase in utilisation of RAS for amniocentesis (8.9% in 2000 to 43.3% of cases in 2006, P < 0.001). CVS RAS was stable. This liberalisation resulted in a fourfold increase in expenditure for FISH and cost/abnormality detected ($A970 per abnormal result in 2000 to $A4015 per abnormal result in 2006). CONCLUSION: The selective use of prenatal RAS results in a reasonably high detection rate for chromosomal anomalies. Liberalisation of RAS, however, is an expensive cytogenetic model. An approach based on some predictive level of risk combined with resource funding levels may be a more pragmatic approach. PMID- 19281577 TI - Body fat composition and weight changes during pregnancy and 6-8 months post partum in primiparous and multiparous women. AB - OBJECTIVE: To compare changes in maternal weight and body fat composition from early to late pregnancy and 6-8 months postnatally between primiparous and multiparous patients. METHODS: Maternal weight and body fat percentage were assessed in a cohort of low-risk uncomplicated women in a general antenatal clinic at 14-20 weeks gestation, after 36 weeks, and around six to eight months after delivery using a Tanita TBF 105 Fat Analyser. Maternal epidemiological and anthropometric data, as well as pregnancy characteristics and perinatal outcome, were derived from standard antenatal records after delivery. The cohort was stratified into primiparous and multiparous women for comparison. RESULTS: In a cohort of 104 women, 55 (52.8%) were primiparous and 49 (47.1%) were multiparous. A relatively good overall correlation between body fat percentage gain and weight gain was observed (correlation coefficient 0.33) from early to late pregnancy. Primiparous women had higher weight gain (12 kg) and higher body fat gain (7.7%) during the pregnancy compared to multiparous women (10.8 kg and 6%, respectively), and they also retained more of the fat accumulated during pregnancy (1.92% vs - 0.44%, P < 0.001) when assessed over six months after their delivery. CONCLUSION: The findings could represent more exaggerated physiological responses to the pregnant state in the primiparous woman as compared to multiparous women. PMID- 19281578 TI - Antenatal screening practice for infectious diseases by general practitioners in Australia. AB - INTRODUCTION: This study aimed to assess self-reported screening practice in the antenatal setting, factors associated with screening, barriers to universal testing for HIV and follow-up for infants born to hepatitis C virus (HCV) infected women. METHODS: A total of 3100 general practitioners (GPs) were mailed the survey. The half from Victoria was randomised to receive their questionnaire by registered post or regular post. All GPs from New South Wales (NSW) received their questionnaire via regular post. RESULTS: The overall response rate was 70%. Registered post resulted in a higher cumulative response rate compared with regular post (86% vs. 67%P < 0.001). Greater than 90% of respondents always screened for syphilis, rubella and hepatitis B virus. Testing for HIV and HCV approached 66% in NSW. In Victoria more respondents always screen for HCV (72%) compared with HIV-1 (64%). Respondents from NSW were less likely to screen for toxoplasmosis (adjusted odds ratio (AOR) 0.64 (0.43, 0.94) P = 0.02) or HCV (AOR 0.75 (0.61, 0.92) P = 0.005) compared with Victoria. Older respondents were more likely to screen for toxoplasmosis (AOR 1.54 (1.05, 2.27) P = 0.03), cytomegalovirus (OR 1.5 (1.0, 2.1) P = 0.05) and chlamydia (AOR 1.88 (1.27, 2.77) P = 0.002). Of respondents who have managed a pregnant woman with HCV 25% inappropriately test infants for infection before one month of age. CONCLUSION: This study highlights the need for more education and resources to increase HIV testing rates and to improve follow-up of an HCV-exposed infant. PMID- 19281579 TI - Early onset, severe fetal growth restriction with absent or reversed end diastolic flow velocity waveform in the umbilical artery: perinatal and long-term outcomes. AB - OBJECTIVE: To assess perinatal and long-term outcomes for pregnancies complicated by early onset, severe fetal growth restriction with absent or reverse end diastolic flow velocity waveform (AREDF) in the umbilical artery. METHODS: A retrospective cohort study of 36 singleton pregnancies with AREDF when the estimated fetal weight (EFW) is less than 501 g at presentation. RESULTS: At presentation, the median gestational age and EFW were 24 (18-29) weeks and 364 (167-496) g, respectively. The median interval between presentation and live birth or diagnosis of intrauterine fetal death (IUFD) was 13 (0-60) days. Delivery was for IUFD in 19 cases (53%), fetal indications in 13 cases (36%) and maternal indications in four cases (11%). Caesarean section (CS) was performed for the 17 live births of which 10 (59%) were by classical CS. Of the total cohort, five infants survived to hospital discharge giving an overall perinatal survival rate of 14%. All survivors had short-term morbidity. The cognitive function in four children was assessed as normal at two years of age. One survivor had developmental delay. None of the surviving children had any evidence of cerebral palsy. CONCLUSION: The overall perinatal survival rate for pregnancies complicated by early onset, severe growth restriction with an EFW of < 501 g and AREDF is low. When delivery occurs for fetal indications, the majority of these women require classical CS. Short-term neonatal morbidity is high though none of the survivors had cerebral palsy. PMID- 19281580 TI - Small for gestational age preterm infants and relationship of abnormal umbilical artery Doppler blood flow to perinatal mortality and neurodevelopmental outcomes. AB - AIM: To determine the outcomes of preterm small for gestational age (SGA) infants with abnormal umbilical artery (UA) Doppler studies. METHODS: A retrospective cohort study of SGA singleton infants delivered between 24 and 32 weeks gestation at King Edward Memorial Hospital, Perth, who had UA Doppler studies performed within seven days of birth. Main outcomes assessed were perinatal mortality and morbidity, and neurodevelopmental outcomes at >or= 1 year of age. Outcomes were compared by normality of UA blood flow. RESULTS: There were 119 infants in the study: 49 (41%) had normal UA Doppler studies, 31 (26%) had an increased systolic diastolic ratio >or= 95th centile, 19 (16%) had absent end diastolic blood flow (AEDF) and 20 (17%) had reversed end-diastolic flow (REDF). Infants in the AEDF and REDF groups were delivered significantly more preterm (P = 0.006) and had lower birthweights (P < 0.001). Ninety four per cent (110 of 117) of live born infants survived. Neurodevelopmental follow-up at 12 months of age or more (median 24 months) was available on 87 of 108 (81%) of live children. Twenty eight per cent (11 of 39) of fetuses who had had AEDF or REDF died or were classified with moderate or severe disability. There was no significant association between abnormality of UA blood flow, perinatal morbidity, perinatal mortality and neurodevelopmental disability after correction for gestational age. CONCLUSION: Fetuses that are SGA with abnormal UA Doppler studies remain at significant risk of perinatal death, perinatal morbidity and long-term neurodevelopmental disability, associated with their increased risk of preterm birth. PMID- 19281581 TI - A comparison of tramadol and pethidine analgesia on the duration of labour: a randomised clinical trial. AB - BACKGROUND: The ideal obstetric analgesia should provide analgesic efficacy without attenuation of uterine contractions. AIMS: To compare the outcome of intramuscular administration of pethidine and tramadol in labour analgesia. METHODS: One hundred and sixty full-term parturients were randomly assigned to two equal groups in active labour. Group P received 50 mg pethidine; and group T, 100 mg tramadol intramuscularly. Primary outcome measure was the duration of the labour. The analgesic efficacy, maternal side-effects, mode of delivery, maternal satisfaction and Apgar score as the secondary outcome were assessed. RESULTS: The duration of labour was shorter in group T, for first stage (190 vs 140 min; P < 0.0001) and for second stage (33 vs 25 min; P = 0.001). There were no differences in Groups P and T with respect to median (7 vs 8) and maximum (7.5 vs 8) visual analog scores (VAS) for pain at 10 min and one hour after drug administration. Women in group P had lower VAS pain scores than those in group T in the second stage of labour (8 vs 9; P = 0/009). There was a significantly higher incidence of nausea and vomiting (35% vs 15%; P = 0.003) and drowsiness (80% vs 29%; P < 0.0001) in group P. CONCLUSION: Both 100 mg tramadol and 50 mg pethidine provide moderate analgesia in first stage of labour. Tramadol seems to cause a shorter duration of labour and lower incidence of maternal side-effects. However, its analgesic efficacy was not found to be as effective as pethidine, especially in the second stage of labour. PMID- 19281582 TI - Obstetric implications of current Australian product information for thyroid related medications. PMID- 19281583 TI - Uterine compression sutures for post-partum bleeding with atony; modification of the B-Lynch suture. AB - We have described a modification of the B-Lynch uterine compression suture which is simple and effective. This technique apposes the anterior and posterior uterine walls, obliterating the cavity and thereby provides effective control of post-partum haemorrhage as a result of uterine atony. It comprises of vertical compression sutures and is distinct from B-Lynch and Haymen's suture by having an additional firm puncture just below the uterine fundus. This means that the suture is transfixed at the uterine fundus, thus eliminating the risk of the sutures sliding off at the uterine fundus. PMID- 19281584 TI - Gynaecologists blaze the trail in primary studies and systematic reviews of diagnostic test accuracy. AB - As the Cochrane Collaboration is poised to begin publishing systematic reviews of diagnostic test accuracy studies in addition to its traditional systematic reviews of treatment effectiveness, we are likely to see a major expansion in the number of primary studies and systematic reviews of diagnostic test accuracy in the medical literature. Obstetricians and gynaecologists have played an important role in initiating this newer area of research. However, the methodology for such studies is challenging and the published literature is riddled with pitfalls. This editorial seeks to simplify the concepts involved in diagnostic test accuracy studies and systematic reviews, to reflect on the early development of this research in our specialty and to envision the future pathway for screening and diagnostic research. PMID- 19281585 TI - Low-dose mifepristone in treatment of uterine leiomyoma: a randomised double blind placebo-controlled clinical trial. AB - AIMS: To evaluate the effect of low-dose mifepristone on leiomyoma-related symptoms, uterine and leiomyoma in women with symptomatic leiomyomata. METHODS: In a double-blind placebo-controlled trial, 40 patients with symptomatic leiomyoma and normal endometrial histology were randomised to receive 10 mg mifepristone (group 1) or placebo (group 2) daily for three months. Leiomyoma related symptoms, uterine, leiomyoma and largest leiomyoma volumes were assessed at baseline and every month for three months. Endometrial biopsy was repeated at the end of therapy. RESULTS: Significant change was noticed between the two groups for mean menstrual blood loss (MBL) by first month. Menstrual blood loss declined by 94.8% in group 1 at three months and 84.2% patients attained amenorrhoea in this group. In group 1 complete relief of dysmenorrhoea occurred in significant number of women (80%) but only 33% patients got rid of pelvic pain. There was no change in these symptoms in group 1 Backache, urinary complaints and dyspareunia were not relieved in either group. Uterine, leiomyoma and largest leiomyoma volume declined by 26-32% in group 1 as compared to none in group 2, and this difference was statistically significant only by the end of the third month of therapy. Mean haemoglobin increased from 9.5 to 11.2 g/dL in group 1. In group 1, at the end of therapy, 63.1% of patients had endometrial hyperplasia without atypia. CONCLUSIONS: Ten milligrams mifepristone for three months is effective in reducing MBL, increasing haemoglobin and reducing uterine and leiomyoma volume with side-effect of endometrial hyperplasia. PMID- 19281586 TI - Survey of surgical skills of RANZCOG trainees. AB - BACKGROUND: In Australia, the Integrated Training Program (ITP) of the Royal Australian and New Zealand College of Obstetricians and Gynaecologists (RANZCOG) offers training in obstetrics and gynaecology. There is anecdotal concern among trainees and Fellows that the surgical component of training is inadequate, with new specialists lacking the confidence and competence to perform many 'standard' surgical procedures. These concerns have not previously been quantified in Australia and New Zealand. AIM: To determine trainees' subjective competence and confidence with surgical procedures and trainees' satisfaction with their surgical training. METHODS: All 430 active RANZCOG trainees and 108 Fellows elevated within the previous two years were invited to complete a self administered questionnaire (65% response rate), which assessed details of procedures performed and confidence to perform them; satisfaction with the surgical training; and perceived teaching ability of the supervising consultants. RESULTS: Those in ITP year 6 rated their confidence high (>or= 4 of 5) for procedures performed very frequently, but lower for other procedures. No procedure regarding the management of complications reached a confidence score of >or= 4. Teaching abilities were rated best for obstetric procedures, with 54% rating their consultants' teaching ability as 'excellent'; but for laparoscopic procedures and procedures dealing with complications, 21.2% and 23.4% of respondents rated their consultants' teaching ability as 'poor', respectively. CONCLUSIONS: Advanced trainees lacked confidence in a range of surgical procedures; and possible weaker areas were identified in the teaching experience of trainers. These limitations must be addressed by medical educators and training program coordinators. PMID- 19281587 TI - HuR expression in the nucleus correlates with high histological grade and poor disease-free survival in ovarian cancer. AB - BACKGROUND: HuR, a nucleo-cytoplasmic shuttling protein, plays an important role in mRNA stability as well as cellular differentiation. Recently, HuR expression, particularly in the cytoplasm, was thought to be associated with the prognosis of several cancers including ovarian cancer. AIMS: To study the clinical significance of nuclear HuR expression in ovarian cancer. METHODS: Primary epithelial ovarian carcinomas (102) and ovarian low malignant potential tumours (11) were assessed for HuR protein expression by immunohistochemistry. HuR scoring accounted for both intensity and percentage of cells stained, and ranged from 0 to 300. RESULTS: HuR was found to be present predominantly in the nucleus, where it was expressed in 85.8% of cases. Nuclear HuR was associated with the invasive cancers (P = 0.004), high grade (P < 0.0001), large residual disease (P = 0.045) and poor disease-free survival (P = 0.0009). Among those 91 specimens with high grade, 76.9% had a high nuclear HuR score, while in those 22 cases with low grade, only 31.8% had a high HuR score (P < 0.0001). Multivariate analysis showed that nuclear HuR intensity was an independent prognostic factor for poor disease-free survival (P = 0.0484). When the invasive cancers were analysed separately, only the association between nuclear HuR and high grade remained (P = 0.0089). CONCLUSIONS: Our results support the clinical significance of nuclear HuR in ovarian carcinoma and suggest that nuclear HuR may also play a role in the biology of ovarian cancer. These data suggest a more complex model for HuR in ovarian cancer than one limited to cytoplasmic localisation. PMID- 19281588 TI - Comparison of TVT and TVT-O in patients with stress urinary incontinence: short term cure rates and factors influencing the outcome. A prospective randomised study. AB - BACKGROUND: Recently, mid-urethral slings have been commonly used in treatment of patients with stress urinary incontinence (SUI). AIMS: To investigate tension free vaginal tape (TVT) and tension-free obturator tape (TVT-O) for surgical treatment of SUI for cure rates (primary endpoint), complications and factors influencing cure rate (secondary endpoints). METHODS: One-hundred and sixty-four patients were included in the study (n = 81 for TVT, n = 83 for TVT-O). The cure rates, complications, preoperative and postoperative urodynamic evaluation, Q-tip test, the Turkish version of Incontinence Impact Questionnaire (IIQ-7) and Urogenital Distress Inventory (UDI-6) scores were recorded. At three and 12 months, the patients were evaluated regarding outcome measures. RESULTS: The cure rates were similar in TVT and TVT-O groups, 88.9% versus 86.7% respectively. Mean operative time was significantly shorter in TVT-O group (P = 0.001). The cure rate was significantly higher in both groups in patients with urethral hypermobility when compared with those with no hypermobility (P = 0.001). CONCLUSIONS: The TVT and TVT-O procedures appear to be equally effective for the treatment of SUI. Also, urethral hypermobility seems to be a factor influencing cure rate of mid-urethral slings. PMID- 19281589 TI - Prevalence of menopausal symptoms and quality of life after menopause in women from South India. AB - OBJECTIVES: This study was carried out to establish the age at onset of menopause and the prevalence of menopause and menopausal symptoms in South Indian women. MATERIALS AND METHODS: Three hundred and fifty-two postmenopausal women attending the outpatient clinics of obstetrics and gynaecology department of Dr TMA Pai Hospital, a tertiary care Hospital in South India, were included in the study. The Menopause-Specific Quality of Life (MENQOL) questionnaire was used in the study. Data were presented as percentages for qualitative variable. RESULTS: The mean age at menopause was 48.7 years. Most frequent menopausal symptoms were aching in muscle and joints, feeling tired, poor memory, lower backache and difficulty in sleeping. The vasomotor and sexual domains were less frequently complained when compared to physical and psychological domains. CONCLUSION: The age at onset of menopause in southern Karnataka (India) is 48.7 years which is four years more than the mean menopause age for Indian women. This could be attributed to better socioeconomic and health-care facility in this region. PMID- 19281590 TI - The early pregnancy assessment project: the effect of cooperative care in the emergency department for management of early pregnancy complications. AB - BACKGROUND: Early pregnancy assessment clinics (EPAC) have been introduced and accepted as the gold standard for management of early pregnancy problems (EPP). However, EPAC are not universally available and management of EPP within the emergency department (ED) can result in prolonged waiting times, inappropriate use of resources and no clear treatment or follow-up plan being implemented. AIM: To assess the effect of an early pregnancy assessment protocol (EPAP) in the ED, designed to create a cultural change among doctors in relation to EPP in order to minimise use of resources, improve treatment times for patients and establish a clear management plan where dedicated EPAC services are not available. METHODS: An intervention, the EPAP was introduced to the ED and retrospective and prospective audits of the patients were carried out to assess the effect. RESULTS: Implementation of the EPAP decreased treatment time by 55%, representations by 48%, pathology blood tests by 56% and formal imaging services by 85%. Gynaecological consultation increased by 37% for each patient visit to the ED and by 9% for each EPP. Total direct cost saving was 63% per patient and no adverse outcomes were recorded. CONCLUSION: Introduction of the EPAP was successful in creating cultural change and delivering clinical and financial benefits to the hospital, patients and staff. Early gynaecological consultation and bedside ultrasound scanning within the ED were key factors. Similar benefits could be reproduced in other institutions and for other clinical scenarios where a need has been identified. PMID- 19281591 TI - Labial fusion causing acute urinary retention in a young adult: a case report. PMID- 19281592 TI - Bioidentical testosterone cream: a rare cause of postmenopausal virilisation. PMID- 19281593 TI - Effect of donor age on the developmental competence of bovine oocytes retrieved by ovum pick up. AB - To study the effect of donor age on oocyte developmental competence and steroid profiles, the crossbred cow (Murray Grey * Brahman) in Yunnan province of China were selected and divided into three groups according to its age. The three groups were young cows (n = 12; 12 months old), middle-aged cows (n = 15; parity: <=3 calvings; age: 7-8 years old) and old cows (n = 10; parity: >=8 calvings; age: >=15 years old). Cumulus-oocyte complexes (COCs) were collected by 10 consecutive ovum pick up (OPU) sessions with a 4-day interval between each session, followed by in vitro maturation, fertilization and embryo development. Results showed that cleavage rates (CR) and blastocyst rates (BR) were higher in the young cows than those in the middle-aged and old cows (p < 0.05). CR and BR from COCs of the first and the fourth OPU sessions were lower than those from other sessions in the young cows and the middle-aged cows (p < 0.05), whereas the similar phenomenon was not observed in the old cows. Plasma concentrations of oestradiol were higher, and plasma concentrations of progesterone were lower before and during OPU sessions in the young cows compared with those in the same period in the middle-aged cows or the old cows (p < 0.01). In conclusion, donor age of oocytes could affect developmental competence of oocytes recovered by OPU through the action of steroid hormonal balance on follicle development. PMID- 19281594 TI - Influence of ovulation status, seasonality and embryo transfer method on development of cloned porcine embryos. AB - To improve pig cloning efficiency, the present study evaluated the effect of ovulation status, seasonality and embryo transfer (ET) method on in vivo development of cloned porcine embryos. Cloned embryos were transferred to surrogate mothers on the same day of somatic cell nuclear transfer. In pre ovulation stage (PO), pregnancy rate (PR) and delivery rate (DR) were 36.3% and 9.4%, respectively. In post-ovulation stage, 22.7% PR and 2.1% DR were recorded (both PR and DR are significantly higher in PO). When ET was performed during winter (December-February), spring (March-May), summer (June-August) and autumn (September-November), the PRs were 13.4%, 37.3%, 24.6% and 51.0%, while DRs were 0%, 12.7%, 4.3% and 7.8%, respectively. The highest PRs were recorded in autumn groups. However, DRs were significantly lower in autumn (7.8%) group compared with spring (12.7%) group. The PR was the lowest and no piglets were born in winter group, which might be because of the effect of low temperature during ET. To overcome the low PR in winter group, 0.25 ml straws were used for ET to minimize exposure time of embryos to ambient temperature. The straw ET group showed significantly higher PR in the winter group (23. 9%) compared with the conventional catheter-loading group (7.7%). We suggest that using PO recipient and ET in spring is the best condition for pig cloning. In addition, alternative method to reduce cold shock during ET in winter is necessary. PMID- 19281595 TI - Morpho-physical recording of bovine conceptus (Bos indicus) and placenta from days 20 to 70 of pregnancy. AB - The study is based on 141 pregnant Bos indicus cows, from days 20 to 70 post insemination. First, special attention was given to the macroscopically observable phenomena of attachment of the conceptus to the uterus, i.e. the implantation, from about days 20 to 30 post-insemination up to day 70, and placentome development by growth, vascularization and increase in the number of cotyledons opposite to the endometrial caruncles. Secondly, as for the conceptuses, semiquantitative, statistical analyses were performed of the lengths of chorio-allantois, amnion and yolk sac; and the different parts of the centre and two extremes of the yolk sacs were also analysed. Thirdly, the embryos/foetuses corresponding to their membranes were measured by their greatest length and by weight, and described by the appearance of external developmental phenomena during the investigated period like neurulation, somites, branchial arcs, brain vesicles, limb buds, C-form, pigmented eye and facial grooves. In conclusion, all the data collected in this study from days 20 to 70 of bovine pregnancy were compared extensively with corresponding data of the literature. This resulted in an 'embryo/foetal age-scale', which has extended the data in the literature by covering the first 8 to 70 days of pregnancy. This age-scale of early bovine intrauterine development provides model for studies, even when using slaughtered cows without distinct knowledge of insemination or fertilization time, through macroscopic techniques. This distinctly facilitates research into the cow, which is now being widely used as 'an experimental animal' for testing new techniques of reproduction like in vitro fertilization, embryo transfer and cloning. PMID- 19281596 TI - Effect of complex vertebral malformation on luteal function in Holstein cows during oestrous cycle and early pregnancy. AB - The reason why cows carrying the mutation of complex vertebral malformation (CVM) show poor reproductive capability although they carry only one mutant allele is still not fully understood. Monitoring the progesterone profiles during oestrous cycle and early pregnancy in carrier cows might help explain their lowered reproductive capability. Progesterone concentration was measured in 19 CVM carrier cows and 21 control cows during oestrous cycle and early pregnancy. Milk samples were collected from all cows starting on the day of artificial insemination until day 45 post-AI. Progesterone was measured in skim milk using enzyme-linked immunosorbent assay (ELISA). Progesterone concentration was significantly reduced on day 7 (p < 0.05) and day 9 (p < 0.01) post-insemination in conceived CVM carrier cows when compared with that in control conceived cows. The mean progesterone concentration during early pregnancy was significantly lower (p < 0.05) in conceived cows with CVM than that of control cows in the same period. However, the mean progesterone concentration did not differ significantly (p = 0.072) in CVM cows that showed fertilization failure or embryonic death than that of control cows. Additionally, of 13 conceived control cows, eight cows (61.5%) showed normal luteal function. In contrast, of nine conceived CVM cows, only four cows (44.4%) showed normal luteal function. The conception rate was 47.4% in CVM carrier cows and 61.9% in control cows, but this difference did not reach significance. In conclusion, progesterone concentration might be lowered during early pregnancy in conceived CVM cows compared with that in control cows. PMID- 19281597 TI - Undernutrition and exogenous melatonin can affect the in vitro developmental competence of ovine oocytes on a seasonal basis. AB - This study evaluated the effects of exogenous melatonin and level of nutrition on oocyte competence, in vitro fertilization (IVF), and early embryonic development in sheep during seasonal anoestrus (SA) and the reproductive season (RS). Adult Rasa Aragonesa ewes were assigned randomly to one of four treatment groups in two experiments based on a 2 x 2 x 2 factorial design. Individuals were treated (+MEL) or not treated (-MEL) with a subcutaneous implant of melatonin for 42 days and then were fed 1.5 (Control, C) or 0.5 (Low, L) times the daily maintenance requirements for 20 days. Ewes were synchronized and mated at oestrus (Day = 0). On Day 5, ovaries were collected and oocytes were used for IVF. Season had a significant (p < 0.01) effect on the number of oocytes recovered (RS: 19.6 +/- 1.0; SA: 14.5 +/- 1.0) and the number of healthy oocytes (RS: 13.9 +/- 0.7; SA: 9.0 +/- 0.7). In the RS, neither nutrition nor melatonin had a significant effect on the evaluated oocytes quality parameters although melatonin implants appeared to reduce the number of unhealthy oocytes in the undernourished group (p < 0.05). During SA, in undernourished ewes exogenous melatonin tended to increase the number of healthy (L+MEL: 9.4 +/- 1.0, L-MEL: 7.6 +/- 1.4; p < 0.1), and significantly improved both cleaved oocytes (L+MEL: 7.0 +/- 0.7, L-MEL: 4.1 +/- 0.9; p < 0.05) and blastocyst rate (L+MEL: 37.2, L-MEL: 21.9%; p < 0.05). In conclusion, oocyte competence in ewes was affected by season, and melatonin implants appeared to improve developmental competence in the seasonal anoestrous period, particularly in experimentally undernourished ewes. PMID- 19281598 TI - Undernutrition during foetal and post-natal life affects testicular structure and reduces the number of Sertoli cells in the adult rat. AB - To test whether undernutrition during foetal to pre-pubertal life would have long lasting effects on testicular histology in adult male offspring, eleven adult Sprague-Dawley pregnant rats were divided into two groups: Control group, n = 4, fed ad libitum, during gestation and lactation (until 25 day post-partum). Underfed group pregnant females (n = 7) were kept in cages where only dams had access to food (standard rat chow, 33.5% of ad libitum intake of Control group pregnant dams). After parturition, litters were adjusted to either 14 (Underfed group) or eight (Control group) pups. Mothers were weighed weekly. At 25 day of age pups were weaned, housed at four animals per cage, fed ad libitum and weighed weekly until euthanized at 100 day of age. Testes were processed for standard histology and morphometrical evaluation. At weaning, mother weight was lower in underfed than in Control group (mean +/- SD): 214.1 +/- 26.2 g vs 361.9 +/- 33.1 g. Body weight at 100 days of age (254 +/- 26.9 g vs 342.4 +/- 10.2 g, p or=1.5 Log10 neutralization index) antibody responses. PMID- 19281605 TI - Capsaicin vs. clobetasol for the treatment of localized alopecia areata. PMID- 19281606 TI - Umbilical sore. PMID- 19281607 TI - Trichodynia is a distinguishing symptom of telogen effluvium. PMID- 19281613 TI - Water vapour permeability of skin care products in relation to molecular and environmental influences. AB - Synopsis An in vitro method for characterization of water retention behaviour of emollients was developed. The human skin is imitated with the aid of a polytetrafluoro ethylene membrane and the degree of water vapour evaporation after impregnation with different emollients is determined. Results obtained from different studies under changing conditions indicate that this is a precise and efficient method which is easy to perform with very simple equipment. It allows the generation of data for the characterization of emollients as an aid for skin care product formulation. Additionally, basic studies on emollient water retention due to exogenous influences are possible. Resume Une methode in vitro pour caracteroen l'aptitude a'la retention d'eau d'un emolliant a ete mise au point. La peau humaine est simulee avec l'aide d'une membrane en polytetrafluoro ethylene (PTFE), et l'evaporation de la voyeur d'eau apres impregnation avec divers emolliants est mesuree. Apres differentes etudes dans diverses conditions experimentales, les resultats obtenus montrent qu'une methode precise et efficaee a ete developpee, facile a methe en ocuore avec un equipement simple. Elle permet d'obtenin les donnecs caracteristiques d'emolliants, facilitant le formulation de produits de soin pour la peau. Des etudes supplementaires concernant l'evolution de la capacitea la retention d'eau d'un emolliant en fonction de facteurs exterieurs sont envisageables. PMID- 19281614 TI - Human stratum corneum as a substrate for in vitro sunscreen testing. AB - Synopsis In this study we have described the construction of a dedicated, inexpensive and portable instrument designed to evaluate sunscreens throughout the UVB and UVA range (290-400 nm). Both Transpore(TM) and an alternative substrate of readily obtainable human stratum corneum have been used as substrates on to which to spread the test products. The transmission of UVR through the substrate of stratum corneum was greater than through Transpore(TM) tape. Both substrates demonstrated a good correlation with in vivo or expected SPF results. However, in 10 out of the 11 sunscreens studied, the comparison of the two substrates demonstrated that the predicted SPF using Transpore(TM) tape was consistently higher than that using human stratum corneum. One sunscreen tested was alcohol based and as such was not suitable to test on Transpore(TM) tape. Predictions of SPF from products with SPF values less than 20 were not significantly different between substrates. However, product SPF values of greater than 20 demonstrated that Transpore(TM) tape overestimated the 'true'SPF. It is postulated that use of human stratum corneum in in vitro SPF testing systems more closely resembles that of human skin in vivo than does Transpore(TM) tape with regard to spreading and absorption of the potential sunscreen product. Resume Cette etude decrit la construction d'un appareil portable bon marche servant aevaluer des ecrans solaires sur les rayons UVA et UVB entre 290 et 400 nm. Le Transpore(TM) et le substrat alternatif d'un stratum corneum ont ete utilises comme substrats sur lesquels on a applique les produits tests. La transmission des rayons UVR a travers le substrat du stratum corneum etait plus importante qu'a travers le Transpores(TM). Chaque substrat a montre une bonne correlation avec les reultats des facteurs de protection solaire prevus ou in vivo. Cependant sur 10 des 11 ecrans solaires etudies, la comparaison des deux substrats a montre que les resultats prevus de facteurs de protection solaires obtenus avec le ruban Transpore(TM)etaient considerablement plus eleves qu'avec le stratum corneum. Un des ecrans solaires teste contenait de l'alcool et ne pouvait etre teste sur le ruban Transpore(TM). Les previsions de SPF avec des produits dont les valeurs SPF etaient inferieures a 20 n'ont pas montre de differences notables entre les substrats. Cependant les valeurs SPF superieures a 20 ont montre que le Transpore(TM) surestimait le SPF reel. Il semble que l'utilisation de stratum corneum humain pour des tests de SPF in vitro corresponde mieux au comportement de la peau humaine in vivo que le Transpore pour l'application et l'absorption d'un ecran solaire potentiel. PMID- 19281615 TI - Optimization of parameters in the emulsification process by two different methods. AB - Synopsis An O/W emulsion with a lipopeptidic emulsifier was optimized by means of two different methods, specifically by experimental design and by selective variation of the parameters. Two optimized emulsions were obtained; they had similar values of emulsification (time and rate of homogenization) and amounts of components. Resume Une emulsion O/E, avec un lipopeptide comme tensioactif, a ete optimisee par deux methodes d'optimisation differentes, notamment par l'experimental design et par la variation selective de certain parametres. Deux emulsions optimisees ont ete ainsi obtenues: ces emulsions ont des valeurs d'emulsification (temp et vitesse d'homogeneisation) et des quantites des composantes tres proches. PMID- 19281616 TI - The effects of surfactants on penetration across the skin*. AB - Synopsis Many of the properties of surfactants can be related to their ability to concentrate at phase interfaces, leading to a reduction in interracial tension. In biological systems the effects of surfactants are complex, particularly their effect on cell and other membranes, and this can lead to alterations in permeability characteristics. This is of particular relevance when considering the stratum corneum which has long been recognized as the major barrier to skin permeation. The magnitude of skin barrier function alteration is dependent on surfactant structure, both the hydrophobic alkyl chain and the hydropnilic ethylene oxide chain demonstrating some structure-activity behaviour. In many biological systems, including skin, surfactants with a similar hydrophilic group will show maximum membrane activity if they possess a decyl or dodecyl alkyl chain. It is difficult to rationalise this phenomenon, given that such solution properties as partition coefficients and CMCs do not show maxima or minima at these chain lengths. It may be that the physical parameters and molecular dimensions of the decyl/dodecyl chain provide the optimal ability to intercalate with the lipid bilayer structure. There is little doubt that once the surfactant has intercalated with the lipid bilayers in the lamellar liquid crystals of the stratum corneum, fluidity in the hydrophobic regions is increased. Effectively, this leads to a looser, more permeable structure. The significance of data obtained using commercially available surfactants, however, can be questioned on the grounds of purity. The purpose of this review is to describe some of the methods used to evaluate the effects of surfactants on the skin barrier and to discuss recent attempts to predict surfactant action on the skin using various biological and physical techniques. Resume La plupart des proprietes des surfactants depend de leur facilitea se concentrer aux interfaces, menant ainsi a la reduction de la tension interfaciale. Dans le cas d'un systeme biologique les effets des surfactants sont complexes, particulierement leur action sur les cellules et les membranes et ceci mene a l'alteration des caracteristiques de la permeabilite. Ce point est tres pertinent pour le stratum corneum qui a ete longtemps considere comme la principale barriere a la penetration cutanee. L'importance de l'alteration de cette function barriere est en relation avec la structure du surfactant. L'importance de son alteration est liee a la fois a la chaine alkyle hydrophobe et a la chaine d'oxyde d'ethylene hydrophile ayant une activite due a sa structure. Dans de nombreux systemes biologiques, incluant la peau, les surfactants avec une chaine hydrophile identique montreront une activite maximum sur les membranes si ils comprennent une chaine alkyle decylique ou dodecylique. Il est difficile de demontrer d'une maniere rationnelle ce phenomene etant donne que les coefficients de partage de telles solutions et les CMC ne permettent pas de determiner des longueurs de chaines. II se peut que les parametres physiques et les dimensions moleculaires des chaines decylique et dodecylique fournissent la capacitt optimale il s'intercaler dans la structure lipidique bicouche. II n'y a pas de doute qu'une fois que le surfactant s'est intercale dans cette bicouche liquide de cristaux lamellaires du stratum corneum, la fluiditt dam les regions hydrophobes est accrue. En effet cela mbne a une structure rellchte plus permtable. La valeur des rtsultats obtenus en utilisant des tensio actifs disponibles sur le march6 peut 6tre remise en question en fonction de leur purete. Le but de cet article est de commenter quelques methodes appliqutes pour cvaluer les effets des surfactants sur la barriere cutante et de discuter differents essais recents pour predire I'action du surfactant sur la peau en utilisant diverses techniques physiques et biologiques. PMID- 19281617 TI - Unnecessary ingredients. PMID- 19281621 TI - Erratum. PMID- 19281618 TI - Influence of zinc oxide in the closure of open skin wounds. PMID- 19281623 TI - Abstracts of the Third Annual London Trauma Conference. London, United Kingdom. November 12-14, 2008. PMID- 19281622 TI - Corrigendum. PMID- 19281624 TI - 12th Joint Meeting of the Signal Transduction Society (STS). Signal Transduction: Receptors, Mediators and Genes Weimar, Germany. 29-31 October 2008. Abstracts. PMID- 19281625 TI - Diet selection and milk production and composition in Girgentana goats with different alpha s1-casein genotype. AB - In goats, alpha s1-casein polymorphism is related to different rates of protein synthesis. Two genetic variants, A and F, have been identified as strong and weak alleles based on a production of 3.5 and 0.45 g/l of alpha s1-casein per allele. The aim of the trial was to test whether goats can select their diet as a function of their genetic aptitude to produce milk at different casein levels and whether this selection can influence milk production or composition. Two groups of 8 animals, homozygous for strong (AA) or weak (FF) alleles were housed in individual pens. Using a manger subdivided into five separate containers, the goats were offered daily for 3 weeks: 1.5 kg of alfalfa pelleted hay, 0.7 kg of whole barley, 0.7 kg of whole maize, 0.7 kg of whole faba bean and 0.7 kg of pelleted sunflower cake. Total dry matter intake was similar between groups and resulted in nutrient inputs much higher than requirements. On average, goats selected 86% of maize plus barley and only 46% of faba bean plus sunflower. Indeed, AA goats selected less faba bean compared with FF goats (37.2 v. 56.7% of the available amount; P=0.01); during week 2 and week 3 they significantly increased maize selection (respectively for week 2 and week 3: 94.9 and 99.1% v. 85.3 and 87.3%) thus increasing the ratio between the high-energy feeds and the high-protein feeds (2.41 v. 1.81, P=0.023). As for true protein, the high soluble fraction (B1) and the indigestible fraction (C) were lower in the diet selected by AA goats (respectively in AA and FF groups: B1, 7.85 v. 9.23% CP, P<0.01; C, 6.07 v. 6.30% CP, P<0.001); these diet characteristics can be associated with lower losses of protein. Milk production, being similar in AA and FF groups when goats were fed with a mixed diet, significantly increased in AA group, when free choice feeding was given (mean productions: 1198 v. 800 g/d, P<0.01). Casein content was higher in AA group than in FF group (2.70 v. 2.40%, P<0.01) whereas milk urea was higher in FF group (59.7 v. 48.8 mg/dl, P<0.01). In conclusion, when the animals were free to select their diet, their higher genetic aptitude to produce casein seemed to adjust their energy and protein dietary input in qualitative terms, thus leading to an increase in milk production and a decrease in milk urea. These results seem to demonstrate that interactions probably occurred between genetic polymorphism at the alpha s1-casein locus, diet selection and the efficiency of nutrient transformation into milk. PMID- 19281626 TI - Relationship between somatic cell count, polymorphonuclear leucocyte count and quality parameters in bovine bulk tank milk. AB - The somatic cell count (SCC) in bovine bulk tank milk is presently used as an indicator of raw milk quality, reflecting the udder health status of the herd. During mastitis, SCC increases, mostly owing to an influx of polymorphonuclear leucocytes (PMN) from blood into milk, with a concomitant change in milk composition. Bulk tank milk samples were categorized according to their SCC, as well as polymorphonuclear leucocyte count (PMNC), to study relationships between SCC, PMNC and various raw milk quality traits, i.e. contents of total protein, whey protein, casein, fat and lactose, casein number, proteolysis and rheological properties. The proportion of PMN, obtained by direct microscopy, was significantly higher in samples with high SCC compared with low SCC samples. SCC and PMNC were strongly correlated, yielding a correlation coefficient of 0.85. High SCC samples had lower lactose and casein contents, lower casein number and more proteolysis than low SCC samples. Samples with high PMNC had a lower casein number than low PMNC samples. Samples with high and low SCC or PMNC did not differ in respect to rheological properties. Our results do not indicate that PMNC is a better biomarker than SCC for raw bulk tank milk quality, as previously proposed. PMID- 19281627 TI - Milk ejection disorders in Swiss dairy cows: a field study. AB - A questionnaire was sent to 2099 dairy farms to investigate the occurrence of poor milkability. Based on that, the frequency of poor milkability in Swiss dairy cows was 4% and the percentage of cows treated with oxytocin (OT) was 2%. In addition, 270 dairy farms that had reported cases of animals with poor milkability were contacted for an interview to classify the disorders. Farmers suspected disturbed milk ejection in 52%, anatomical dysfunction of the teat and/or the udder in 16% and milk ejection disorder or impaired milkability caused by discernable environmental factors in 32% of the cases. Forty-eight animals from 18 farms with suspected milk ejection disorders were selected for an experimental field study which included milk flow recording and OT administration to induce milk ejection. After cessation of the spontaneous milk flow, a low dose of OT (0.2, 0.5 or 1 i.u.) was injected i.v. to test the responsiveness of the udder to OT at a physiological level. When milk flow ceased again, 10 i.u. OT was injected i.v. (supraphysiological) to ensure complete udder emptying and to determine the residual milk. Milk ejection disorder could be confirmed in 69% of the cases, i.e. if residual milk was >20% of the total milk. Because in 27% of the animals milk ejection disorder was not confirmed on the basis of elevated residual milk, an anatomical disorder of the teat and/or the udder was suspected. Milk ejection disorder could be confirmed in 69% of the cases whereas in 27% of the suspected cases an anatomical disorder of the teat and/or the udder was suspected. An increased cortisol production in cows with milk ejection disorder was not obvious because faecal concentrations of cortisol metabolites with a 5 beta-androstane-3 alpha,11 oxo-structure were not augmented in animals with disturbed milk ejection. PMID- 19281628 TI - Impact of policy-induced structural change on milk quality: evidence from the Flemish dairy sector. AB - This paper uses a Markov chain model to analyse the dynamics in farm-size distribution among the Flemish dairy sector and the impact of quota policy regulation on such changes. The model predicts a decline of 24% in number of farms in 2014 compared with the current situation with a more liberal exchange policy and a decline of 18% with a restricted quota exchange policy. From these Markov chain model results, we analysed the impact of farm-size distribution on eight different milk quality parameters (total bacterial count, somatic cell count, coliform count, freezing point, urea-N, fat content and protein content and penalty-points). In general, larger farms produce higher quality milk than smaller farms, especially with respect to the microbiological parameters (total bacterial count, somatic cell count and coliform count). The change in farm-size distribution from a liberal quota exchange policy would decrease the average total bacterial count by 18.0%, the somatic cell count by 2.1% and the coliform count by 11.0%. The aggregate performance of the other parameters are smaller with improvements in all cases of <1%. PMID- 19281629 TI - Effect of sunflower-seed oil or linseed oil on milk fatty acid secretion and lipogenic gene expression in goats fed hay-based diets. AB - Plant oils in the diet are known to alter milk fat composition owing to changes in the supply of fatty acid precursors and/or activity of lipogenic enzymes in the mammary gland. Thirteen mid-lactating Alpine goats were used in a 3 x 3 Latin square design with 28-d periods to evaluate possible mechanisms regulating milk fat synthesis and fatty acid composition on grass hay-based diets containing none (H) or 55 g/kg diet dry matter of sunflower-seed oil (HSO) or linseed oil (HLO). Inclusion of oils in the diet had no effect on milk yield but enhanced (P<0.05) milk fat secretion. Compared with the control, HLO and HSO decreased (P<0.05) C10 C16 secretion and increased (P<0.05) C18 output in milk, responses that were accompanied by reductions in milk fat cis-9 14:1/14:0, cis-9 18:1/18:0 and cis-9, trans-11 18:2/cis-9 18:1 concentration ratios. Plant oil supplements decreased (P<0.05) mammary stearoyl-CoA desaturase (SCD) activity but had no effect on SCD mRNA. Treatments had no effect on glucose-6-phosphate dehydrogenase, malic enzyme and glycerol-3-phosphate dehydrogenase activity, or mRNA abundance and/or activity of lipoprotein lipase, acetyl-CoA carboxylase and fatty acid synthase in mammary, hepatic or adipose tissue. The results provided little support for milk fatty acid secretion responses to HLO and HSO being mediated via changes in mammary, hepatic or adipose mRNA abundance or in the activity of key lipogenic enzymes. In conclusion, plant oils in the diet enhance milk fat synthesis, alter milk fatty acid composition and specifically inhibit mammary SCD activity in the goat. Furthermore, the results suggest that the regulation of mammary lipogenesis in response to plant oils appears related to factors other than altered mammary gene expression or potential lipogenic enzyme activity. PMID- 19281630 TI - A compartmental model describing changes in progesterone concentrations during the oestrous cycle. AB - The objective of this study was to develop a mathematical model that accurately describes the rise and decline in plasma progesterone concentrations, and is able to define parameters describing progesterone appearance and disappearance during the bovine oestrous cycle. Daily plasma progesterone data from 27 cows were used to develop a compartmental model consisting of an appearance function and an appearance modulating function. Model outputs included an apparent appearance or secretion duration, appearance rate and an average disappearance rate (expressed as arbitrary units per day; units/d). Shape-based clustering identified three common shape-based groups (or clusters) of progesterone profiles defined as either 'peaked' profile, with the profile reaching a distinguishable peak, 'structured', with the profile exhibiting a wave-like pattern, or 'flat top', with the profile reaching a plateau. Differences in the model parameters for the three different shapes of progesterone profiles were examined: peaked (n=13), flat top (n=7) and structured (n=7). The mean duration of apparent appearance was 11.49 (SD 0.17 d) for all 27 profiles. The model estimates for total appearance of progesterone (area under the curve; ng/ml per cycle), mean appearance rate and maximum appearance rate were 69.04 ng/ml per cycle (SD 15.2 ng/ml per cycle), 3.19 ng/ml per cycle (sd 0.7 ng/ml per d) and 6.70 ng/ml (SD 1.31 ng/ml), respectively. The average disappearance rate was 1.0 units/d (SD 0.04 units/d). The apparent appearance duration was greatest (P<0.01) in the flat top profiles (12.54, SD 0.41 d) followed by the structured (11.77, SD 0.66 d) and the peaked (10.80, SD 0.30 d) profiles. Total and mean progesterone appearance, maximum progesterone appearance rate, and the progesterone disappearance rates were not different between the profiles. The model successfully simulated all components of the progesterone profile and was able to define specific parameters of different shaped progesterone profiles. A simple model able to estimate parameters describing progesterone appearance and disappearance can be used to explore the relationships between profile shapes and reproductive outcomes. PMID- 19281631 TI - Safety assessment of Lactobacillus fermentum CECT5716, a probiotic strain isolated from human milk. AB - Lactobacillus fermentum CECT5716, a probiotic strain isolated from human milk, was characterized in a previous study. The objective of this study was to evaluate its sensitivity to antibiotics and its potential toxicity and translocation ability after oral administration to mice. For this purpose, 40 Balb/C mice were divided in two groups (n=20 per group). One group was treated orally with 10(10) colony forming units (cfu)/mouse/day of Lb. fermentum CECT5716 during 28 d. The other group only received the excipient and was used as control. Food intake, body weight, bacterial translocation and different biochemical and haematological parameters were analysed. Oral administration of Lb. fermentum CECT5716 to mice had no adverse effects on mice. There were no significant differences in body weight or food intake between control and probiotic-treated mice. No bacteraemia was observed and there was no treatment-associated bacterial translocation to liver or spleen. Stress oxidative markers were not different in control and probiotic-treated mice. These results suggest that the strain Lb. fermentum CECT5716 is non-pathogenic for mice even in doses 10,000 times higher (expressed per kg of body weight) than those normally consumed by humans. PMID- 19281632 TI - Application of a multiplex PCR assay for the detection of Shigella, Escherichia coli and Shiga toxin-producing Esch. coli in milk. AB - A multiplex PCR (mPCR) assay using previously known genetic markers of Shigella, Escherichia coli and Shiga-toxic Esch. coli was standardized. uidA gene was targeted for the common detection of Esch. coli and Shigella, whereas ipaH and stx1 genes were used as markers for the detection of Shigella and shiga-toxin producing strains, respectively. The standardized assays detected the target organism specifically and selectively. The mPCR developed by combining all the three reactions generated specific products. The inclusivity and exclusivity tests depicted the precise specificity of the mPCR assay. Results were interpreted on the basis of the pattern of amplicons generated: amplifications of the ipaH and uidA gene fragments indicated the presence of Shigella spp., amplification of uidA alone revealed the presence of Esch. coli and additional presence of verotoxin gene amplicon indicated verotoxinogenic nature of the strain. Specific patterns of bands were obtained when different strains of Esch. coli and Shigella spp. were subjected to this assay. The reactions, individually as well as in the mPCR, could detect approximately 1 cell per 20-microl PCR assay. The protocols were validated by analyzing the coded samples of full fat milk spiked with different pathogens. In naturally contaminated raw milk samples (n=100), Esch. coli were detected in all samples and verotoxinogenic Esch. coli in 15 samples. Shigella, however, was not detected in any of the samples. When DNA purified from the samples found positive for Shiga-toxic Esch. coli was directly used as template for the mPCR, the results showed agreement with the enrichment based detection. The mPCR assay, standardized in this study, may be used for rapid microbiological evaluation of milk samples. Further, the study emphasizes the need for better hygienic conditions in dairies. PMID- 19281633 TI - Effect of somatic cell counts on ewes' milk protein profile and cheese-making properties in different sheep breeds reared in Spain. AB - Bulk tank ewe's milks from Assaf, Castellana and Churra breeds categorized within three different Somatic Cell Count (SCC) groups (LSCC: <500,000; MSCC: 1,000,000 to 1,500,000; and HSCC: 2,500,000 to 3,000,000 cells ml-1) were used to investigate changes in capillary electrophoresis protein profiles and cheese making properties. The results do not reveal a significant effect of SCC on total casein contents, because the sum of beta-caseins decreased as SCC increased; no statistically significant differences were observed for the sum of alpha-caseins, and the values of kappa-casein were higher in the HSCC milk. However, the soluble proteins other than alpha-lactalbumin and beta-lactoglobulin increased with SCC. Regarding the effect of breed, the Assaf breed had the lowest contents of kappa CN, alpha s1-I-CN, alpha s1-II-CN 1-CN, alpha s1-III-CN, beta1-CN and beta2-CN. The protein profile was significantly correlated with curd textural properties. alpha s1-I-CN was the most influential variant because it was positively correlated with a large number of textural parameters. Cheese yield was positively correlated with all casein variants except alpha s1-III-CN, showing that the milk from local breeds were more suitable for cheese-making due to their higher contents of all the casein variants. Regarding curd texture properties LSCC milk curds showed more cohesiveness, associated with its lower content of alpha s1-III-CN and Castellana milk curds showed the highest values for firmness owing to their higher content of alpha s1-I-CN. PMID- 19281634 TI - Enumeration of starter cultures during yogurt production using Petrifilm AC plates associated with acidified MRS and M17 broths. AB - The efficiency of Petrifilm AC (3M Microbiology, St. Paul, MN, USA) associated with the broths M17 and de Mann-Rogosa-Sharpe (MRS) at pH 5.4 was evaluated to enumerate Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus during the yogurt production. Commercial and reference strains of these microorganisms were experimentally inoculated in nonfat milk and incubated at 42 degrees C for 4 h for yogurt production. At the moment of inoculation and after incubation, aliquots were collected, submitted to dilution using the broths M17 and MRS at pH 5.4, and plated for Strep. salivarius and Lb. bulgaricus enumeration according ISO 9232 and at Petrifilm AC plates, respectively. M17 plates were incubated at 42 degrees C, and MRS plates were incubated at 35 degrees C under anaerobiosis. After 48 h, the formed colonies were enumerated and the counts were compared by correlation and analysis of variance (P<0.05). In addition, colonies were randomly selected from all plates and characterized according to Gram staining and morphology. The obtained results indicated that Petrifilm AC plates associated to M17 and MRS at pH 5.4 can be considered as a suitable alternative for Strep. salivarius and Lb. bulgaricus enumeration during yogurt production, with slight interferences due to the acidity of MRS at the moment of inoculation, and due to the acidity of yogurt at the end of fermentation process. It was also observed that the MRS at pH 5.4 was not sufficiently selective for Lb. delbrueckii enumeration, despite it is indicated by the official protocol from ISO 9232. PMID- 19281635 TI - Twelve-month-olds learn novel word-object pairings differing only in stress pattern. AB - Infants at 1 ; 2 demonstrate difficulty in accessing subtle phonetic information about newly learned word-object pairings (Stager & Werker, 1997). In this study, we examined whether or not infants can access subtle prosodic information such as lexical stress in a word learning task. We tested infants younger than 1 ; 2 to see if they could learn two new word-object associations that differ only in stress pattern (Sww versus wSw). Our results are the first to demonstrate that, even without contextual support, infants at 1 ; 0 succeed at this task, suggesting that the salient acoustic properties associated with lexical stress facilitate word-object associative learning. PMID- 19281636 TI - The effect of single dose versus two doses of praziquantel on Schistosoma haematobium infection and pathology among school-aged children in Mali. AB - The aim of this study was to assess the effect of two doses of 40 mg/kg praziquantel with 2 weeks interval versus a standard single dose of 40 mg/kg on cure rates, egg reduction, intensity of infection, and micro-haematuria in Schistosoma haematobium infections. A randomised controlled intervention study was carried out among school-aged children in two different endemic settings with follow-up at 3, 6 and 18 months following drug administration. Differences in cure rates between the two treatment regimens were not significant. However, in high transmission areas, the double treatment regimen was more effective in egg reduction than single treatment regimen and the difference in egg reduction between the two treatments was significant at 3 months (P<0.005), 6 months (P<0.0001) and 18 months (P<0.003) after treatment. There was a significant difference in the effect of the two treatments on prevalence of micro-haematuria at 18-month follow-up in both Koulikoro (P<0.001) and Selingue (P<0.003). The study shows that although no significant difference could be observed in the overall cure-rates between the two treatment regimens, the effect of double treatment was a significant reduction in infection intensity as well as micro haematuria which may have a great impact in reducing subtle morbidity. PMID- 19281637 TI - Praziquantel: its use in control of schistosomiasis in sub-Saharan Africa and current research needs. AB - Treatment with praziquantel (PZQ) has become virtually the sole basis of schistosomiasis control in sub-Saharan Africa and elsewhere, and the drug is reviewed here in the context of the increasing rate that it is being used for this purpose. Attention is drawn to our relative lack of knowledge about the mechanisms of action of PZQ at the molecular level, the need for more work to be done on schistosome isolates that have been collected recently from endemic areas rather than those maintained in laboratory conditions for long periods, and our reliance for experimental work mainly on Schistosoma mansoni, little work having been done on S. haematobium. There is no evidence that resistance to PZQ has been induced in African schistosomes as a result of its large-scale use on that continent to date, but there is also no assurance that PZQ and/or schistosomes are in any way unique and that resistant organisms will not be selected as a result of widespread drug usage. The failure of PZQ to produce complete cures in populations given a routine treatment should therefore solicit considerable concern. With few alternatives to PZQ currently available and/or on the horizon, methods to monitor drug-susceptibility in African schistosomes need to be devised and used to help ensure that this drug remains effective for as long a time as possible. PMID- 19281638 TI - Naphthoimidazoles promote different death phenotypes in Trypanosoma cruzi. AB - SUMMARY: In a screening of 65 derivatives of natural quinones using bloodstream trypomastigotes of Trypanosoma cruzi, the 3 naphthoimidazoles derived from beta lapachone - N1, N2 and N3--were selected as the most active. Investigation of their mode of action led to the characterization of mitochondrion, reservosomes and DNA as their main targets, and stimulated further studies on death pathways. Ultrastructural analysis revealed both autophagic (autophagosomes) and apoptotic like (membrane blebbing) phenotypes. Flow cytometry analysis showed, in N2 treated trypomastigotes, a small increase of phosphatidylserine exposure, and a large increase in the percentage of necrosis, caused by N1 or N2. These death phenotypes were not detected in treated epimastigotes. The strong increase in labelling of monodansyl cadaverine, the inhibition of the death process by wortmannin or 3-methyladenine, the overexpression of ATG genes in treated epimastigotes, together with ultrastructural evidence point to autophagy as the predominant phenotype induced by the naphthoimidazoles. However, there are other pathways occurring concomitantly with variable intensities, justifying the need to detail the molecular features involved. PMID- 19281639 TI - Schistosoma mansoni TOR is a tetraspanning orphan receptor on the parasite surface. AB - SUMMARY: A trispanning orphan receptor (TOR) has been described in Schistosoma haematobium and S. mansoni. Here we report the complete molecular organization of the S. mansoni TOR gene, also known as SmCRIT (complement C2 receptor inhibitor trispanning). The SmTOR gene consists of 4 exons and 3 introns as shown by cloning the single exons from S. mansoni genomic DNA and the corresponding cDNA from the larval stage (cercaria) and the adult worm. The SmTOR ORF consists of 1260 bp and is longer than previously reported, with a fourth trans-membrane domain (proposed new name: Tetraspanning Orphan Receptor) and with, however, an unchanged C2-binding domain on the extracellular domain 1 (ed1). This domain differs in S. japonicum. A protein at the approximate expected molecular weight (55 kDa) was detected in adult worm extracts with polyclonal and monoclonal antibodies, and was found to be expressed on the tegumental surface of cercariae. PMID- 19281641 TI - Systematic method for the kinetic modeling of temporally resolved hyperspectral microscope images of fluorescently labeled cells. AB - In this paper we report the application of a novel method for fitting kinetic models to temporally resolved hyperspectral images of fluorescently labeled cells to mathematically resolve pure-component spatial images, pure-component spectra, and pure-component reaction profiles. The method is demonstrated on one simulated image and two experimental cell images, including human embryonic kidney cells (HEK 293) and human A549 pulmonary type II epithelial cells. In both cell images, inhibitor kappa B kinase alpha (IKK(alpha)) and mitochondrial antiviral signaling protein (MAVS) were labeled with green and yellow fluorescent protein, respectively. Kinetic modeling was performed on the compressed images by using a separable least squares method. A combination of several first-order decays were needed to adequately model the photobleaching processes for each fluorophore observed in these images, consistent with the hypothesis that each fluorophore was found in several different environments within the cells. Numerous plausible mechanisms for kinetic modeling of the photobleaching processes in these images were tested and a method for selecting the most parsimonious and statistically sufficient model was used to prepare spatial maps of each fluorophore. PMID- 19281642 TI - Hyperspectral confocal fluorescence imaging: exploring alternative multivariate curve resolution approaches. AB - Hyperspectral confocal fluorescence microscopy, when combined with multivariate curve resolution (MCR), provides a powerful new tool for improved quantitative imaging of multi-fluorophore samples. Generally, fully non-negatively constrained models are used in the constrained alternating least squares MCR analyses of hyperspectral images since real emission components are expected to have non negative pure emission spectra and concentrations. However, in this paper, we demonstrate four separate cases in which partially constrained models are preferred over the fully constrained MCR models. These partially constrained MCR models can sometimes be preferred when system artifacts are present in the data or where small perturbations of the major emission components are present due to environmental effects or small geometric changes in the fluorescing species. Here we demonstrate that in the cases of hyperspectral images obtained from multicomponent spherical beads, autofluorescence from fixed lung epithelial cells, fluorescence of quantum dots in aqueous solutions, and images of mercurochrome-stained endosperm portions of a wild-type corn seed, these alternative, partially constrained MCR analyses provide improved interpretability of the MCR solutions. Often the system artifacts or environmental effects are more readily described as first and/or second derivatives of the main emission components in these alternative MCR solutions since they indicate spectral shifts and/or spectral broadening or narrowing of the emission bands, respectively. Thus, this paper serves to demonstrate the need to test alternative partially constrained models when analyzing hyperspectral images with MCR methods. PMID- 19281643 TI - Third-harmonic generation and multi-photon excitation fluorescence imaging microscopy techniques for online art conservation diagnosis. AB - We present an appropriate methodology and results for using third-harmonic generation (THG) modality for nondestructive high resolution imaging measurements of varnished structures in model painted artifacts. Detection takes place in the reflection mode, demonstrating the ability of the technique to be applied to the evaluation of original artworks. Furthermore, multi-photon excitation fluorescence images were obtained, providing complementary information related to the identification of the chemical composition of the artifacts. PMID- 19281644 TI - Transcutaneous Raman spectroscopy of murine bone in vivo. AB - Raman spectroscopy can provide valuable information about bone tissue composition in studies of bone development, biomechanics, and health. In order to study the Raman spectra of bone in vivo, instrumentation that enhances the recovery of subsurface spectra must be developed and validated. Five fiber-optic probe configurations were considered for transcutaneous bone Raman spectroscopy of small animals. Measurements were obtained from the tibia of sacrificed mice, and the bone Raman signal was recovered for each probe configuration. The configuration with the optimal combination of bone signal intensity, signal variance, and power distribution was then evaluated under in vivo conditions. Multiple in vivo transcutaneous measurements were obtained from the left tibia of 32 anesthetized mice. After collecting the transcutaneous Raman signal, exposed bone measurements were collected and used as a validation reference. Multivariate analysis was used to recover bone spectra from transcutaneous measurements. To assess the validity of the transcutaneous bone measurements cross-correlations were calculated between standardized spectra from the recovered bone signal and the exposed bone measurements. Additionally, the carbonate-to-phosphate height ratios of the recovered bone signals were compared to the reference exposed bone measurements. The mean cross-correlation coefficient between the recovered and exposed measurements was 0.96, and the carbonate-to-phosphate ratios did not differ significantly between the two sets of spectra (p > 0.05). During these first systematic in vivo Raman measurements, we discovered that probe alignment and animal coat color influenced the results and thus should be considered in future probe and study designs. Nevertheless, our noninvasive Raman spectroscopic probe accurately assessed bone tissue composition through the skin in live mice. PMID- 19281645 TI - Reducing inter-replicate variation in fourier transform infrared spectroscopy by extended multiplicative signal correction. AB - Fourier transform infrared (FT-IR) spectroscopy is a powerful tool for characterizing biological tissues and organisms, but it is plagued by replicate variation of various sources. Here, a method for estimating and correcting unwanted replicate variation in multivariate measurement signals, based on extended multiplicative signal correction (EMSC), is presented. Systematic patterns of unwanted methodological variations are estimated from replicate spectra, modeled by a linear subspace model, and implemented into EMSC. The method is applied to FT-IR spectra of two different sets of microorganisms (different double gene knockout strains of Saccharomyces cerevisiae and different species of Listeria) and compared to other preprocessing methods used in FT-IR absorption spectroscopy of microorganisms. The EMSC replicate correction turns out to perform best among the compared methods. PMID- 19281646 TI - Application of time-of-flight near-infrared spectroscopy to fruits: analysis of absorption and scattering conditions of near-infrared radiation using cross correlation of the time-resolved profile. AB - The absorption and scattering conditions of near-infrared radiation in a grapefruit, a popular thick-peeled fruit, were investigated by time-of-flight near-infrared spectroscopy (TOF-NIRS). The cross-correlation function was introduced to obtain fine spectroscopic information from the time-resolved profile. Variation of the optical parameters in both the time-resolved profile and the cross-correlation function showed that the NIR radiation was largely absorbed in the peel and considerably scattered in the flesh of the fruit. It also reflected the straightness of the input pulsed laser. The substantial optical path length of the grapefruit estimated from the cross-correlation function was approximately 4 to 5 times as long as the nominal optical path length (NOPL). The cross-correlation function was an effective tool to analyze the absorption/scattering conditions of NIR radiation in a sample where an unstable light source such as a Nd:YAG laser with high output energy was employed. PMID- 19281647 TI - Preliminary investigations into macroscopic attenuated total reflection-fourier transform infrared imaging of intact spherical domains: spatial resolution and image distortion. AB - This paper describes preliminary investigations into the spatial resolution of macro attenuated total reflection (ATR) Fourier transform infrared (FT-IR) imaging and the distortions that arise when imaging intact, convex domains, using spheres as an extreme example. The competing effects of shallow evanescent wave penetration and blurring due to finite spatial resolution meant that spheres within the range 20-140 microm all appeared to be approximately the same size ( approximately 30-35 microm) when imaged with a numerical aperture (NA) of approximately 0.2. A very simple model was developed that predicted this extreme insensitivity to particle size. On the basis of these studies, it is anticipated that ATR imaging at this NA will be insensitive to the size of intact highly convex objects. A higher numerical aperture device should give a better estimate of the size of small spheres, owing to superior spatial resolution, but large spheres should still appear undersized due to the shallow sampling depth. An estimate of the point spread function (PSF) was required in order to develop and apply the model. The PSF was measured by imaging a sharp interface; assuming an Airy profile, the PSF width (distance from central maximum to first minimum) was estimated to be approximately 20 and 30 microm for IR bands at 1600 and 1000 cm( 1), respectively. This work has two significant limitations. First, underestimation of domain size only arises when imaging intact convex objects; if surfaces are prepared that randomly and representatively section through domains, the images can be analyzed to calculate parameters such as domain size, area, and volume. Second, the model ignores reflection and refraction and assumes weak absorption; hence, the predicted intensity profiles are not expected to be accurate; they merely give a rough estimate of the apparent sphere size. Much further work is required to place the field of quantitative ATR-FT-IR imaging on a sound basis. PMID- 19281648 TI - Cotton fiber quality characterization with light scattering and fourier transform infrared techniques. AB - Fiber quality measurement is critical to assessing the value of a bale of cotton for various textile purposes. An instrument that could measure numerous cotton quality properties by optical means could be made simpler and faster than current fiber quality measurement instruments, and it might be more amenable to on-line measurement at processing facilities. To that end, a laser system was used to investigate cotton fiber samples with respect to electromagnetic scattering at various wavelengths, polarization angles, and scattering angles. A Fourier transform infrared (FT-IR) instrument was also used to investigate the transmission of electromagnetic energy at various mid-infrared wavelengths. Cotton samples were selected to represent a wide range of micronaire values. Varying the wavelength of the laser at a fixed polarization resulted in little variation in scattered light among the cotton samples. However, varying the polarization at a fixed wavelength produced notable variation, indicating that polarization might be used to differentiate among cotton samples with respect to certain fiber properties. The FT-IR data in the 12 to 22 microm range produced relatively large differences in the amount of scattered light among all samples, and FT-IR data at certain combinations of fixed wavelengths were highly linearly related to certain measures of cotton quality including micronaire. PMID- 19281649 TI - Mid-infrared trace gas analysis with single-pass fourier transform infrared hollow waveguide gas sensors. AB - A hollow core optical fiber gas sensor has been developed in combination with a Fourier transform infrared (FT-IR) spectrometer operating in the spectral range of 4000-500 cm(-1), enabling continuous detection of small volume gas-phase analytes such as CH(4), CO(2), C(2)H(5)Cl, or their mixtures at trace levels. Ag/Ag-halide hollow core optical fibers simultaneously serve as an optical waveguide for broad-band mid-infrared radiation and as a miniaturized absorption gas cell. Specifically, carbon dioxide, methane, and ethyl chloride as well as binary mixtures in a carrier gas were analyzed during exponential dilution experiments. In the studies reported here, the integration of an optical gas sensor with FT-IR spectroscopy provides excellent detection limits for small gas volumes ( approximately 1.5 mL) of individual analytes at a few tens of parts per billion (ppb, vol/vol) for carbon dioxide and a few hundreds of ppb (vol/vol) for methane. Furthermore, the broad-band nature of the radiation source and of the hollow core optical waveguide provides the capability of multi-constituent analysis in mixtures. PMID- 19281650 TI - Improved algorithm for quantitative analyses of infrared spectra of multicomponent gas mixtures with unknown compositions. AB - We present a major improvement of an algorithm based on a spectral library search for the quantitative analysis of multicomponent gas samples with unknown compositions. A quantitative spectral database of infrared spectra is used as a training set to compute regression coefficients. Concentrations are computed in the principal component space via principal component regression (PCR). In addition to previous algorithms, we introduce a rating for each candidate substance depending on the concentration found with PCR and a filter that removes candidates that are predicted with negative concentrations if their rating is below a certain threshold. Negative concentrations arise when the measured spectrum contains components that are not contained in the database. The PCR is recomputed until all candidates have a rating above the threshold. Then an adaptive filter "subtracts" the substance with the highest rating from both the measured spectrum and the library and appends it to a hit list. The iteration of this procedure directly produces a list of substances in order of descending importance (i.e., contribution to the measured absorbance) with their corresponding concentrations. The algorithm is tested on spectra of multicomponent surgical smoke samples. The four main components (water, methane, ethane, and ethene) are identified correctly (within the top 5 of the hit list) for an appropriate choice of the rating threshold. The algorithm describes the composition of the smoke sample correctly despite the presence of features in the spectrum that cannot be explained by the spectrum of any single substance present in the database. PMID- 19281651 TI - An insight into sequential order in two-dimensional correlation spectroscopy. AB - Determination of the sequential order of events is a very important feature of generalized two-dimensional correlation spectroscopy (2D-COS). Recently, queries have been put forward on the actual effectiveness of this method when changes are non-periodic, particularly in the presence of local sequential order. Consequently, it brings an urgent necessity to understand the true connotation of the sequential order parameter derived from 2D-COS analysis. This article presents an attempt to address these questions based on the analysis of simulated spectra by assuming the band intensity changes in a logarithmic, exponential, hyperbolic, or polynomial manner. It is concluded that for two events changing monotonically and without local sequential orders, one event occurring prior to the other as determined using 2D-COS means in most cases that the former has both a shorter half-time and a greater half-intensity. As a rule of thumb, intensity versus the perturbation factor should be plotted before performing the 2D-COS analysis to determine the sequential order of the involved events. In the presence of obvious local sequential order, 2D-COS analysis is unnecessary. Otherwise, sequential order can be determined quite reliably based on 2D-COS analysis. PMID- 19281652 TI - Near-infrared spectroscopy as a useful tool for analysis in solution in common organic solvents. AB - In the present paper we report that near-infrared spectroscopy is a useful tool for analyzing solutes in solution in common organic solvents. This is because the near-infrared absorptions of the solvent are not so strong as to disturb the separation of the characteristic near-infrared bands, by subtraction, of the solute. To demonstrate this capability, we first showed that the near-infrared absorptions of heptane and toluene, each of which represents aliphatic or aromatic solvents, do not significantly affect the noise level of the difference spectrum, in which the near-infrared spectrum of a solute is to be separated by subtraction. Second, we showed that the characteristic near-infrared absorptions of 1-heptene as a solute were well separated from the spectrum of the solution in heptane and toluene. Four of the five indicator bands of 1-heptene were recognizable and sufficiently detected at the almost limiting concentration of 0.1% (v/v) in both solvents. The minimum magnitude of the detectable signal is discussed in terms of the signal-to-noise ratios of the indicator bands. As an application, we investigated the interaction properties of a C identical with CH group of 1-heptyne from the CH stretching fundamental, combination, and overtone bands in the pure liquid and in solution in three solvents at various concentrations. PMID- 19281653 TI - Collisional quenching of erythrosine B as a potential reference dye for impulse response function evaluation. AB - We studied the collisional quenching of the erythrosine B fluorophore by potassium iodide. The quenching follows a Stern-Volmer dependence up to the highest quencher concentration. The lifetime of erythrosine B decreases to 24 ps in 5.02 M of potassium iodide. The quantum yield of erythrosine B in the presence of 5.02 M KI is 0.0035. The relatively high brightness makes this compound attractive as an ultrashort reference in time-resolved measurements. In both frequency- and time-domain fluorescence techniques, there is a need for lifetime standards with extremely short decay times. Mimicking the instantaneous scattering at longer wavelengths allows color-effect-free measurements in the emission region. Another motivation is the problem of obtaining the impulse response function in the case of two-photon excitation. Time-resolved microscopy also benefits from fast-decaying dyes because the impulse response function can be evaluated at the emission wavelength of the investigated specimen without changing filters. We demonstrated that impulse response functions for commonly used detectors are practically the same for scattering as for quenched erythrosine B emission. We also analyzed a complex fluorescence decay using both elastic scattering and quenched erythrosine B emission as a response function. PMID- 19281654 TI - A new method for performing polarization modulation infrared reflection adsorption spectroscopy of surfaces. AB - A new and relatively simple polarization modulation technique is presented and tested that enables the whole spectral range to be detected between 400 and 4000 cm(-1). This experiment is conventionally carried out using a photoelastic modulator that modulates incident plane polarized light through 90 degrees . This suffers from the drawback that it enables spectra to be collected only over a relatively narrow spectral range. As an alternative, a polarizer is placed in the beam and oriented at 45 degrees to the sample normal. This produces incident radiation fluxes with identical intensities for both s- and p-polarized light. A second polarizer is then modulated through 90 degrees and the surface spectrum is then extracted in the usual manner from the difference between these signals, normalized to their sum. The method is demonstrated for a self-assembled monolayer of 11-mercapto-undecanoicacid (11-MUA) on gold on mica, and it is shown that, while the resulting spectra are extremely sensitive to optical alignment, the method yields spectra that are in excellent agreement with published data. PMID- 19281655 TI - Novel micro-cavity substrates for improving the Raman signal from submicrometer size materials. AB - A novel and simple method for improving the detection limit of conventional Raman spectra using a micro-Raman system and picoliter volumes is presented. A micro cavity in a reflecting metal substrate uses various mechanisms that collectively improve the entire Raman spectrum from the sample. A micro-cavity with a radius of several micrometers acts as a very effective device that provides multiple excitation of the sample with the laser and couples the forward-scattered Raman photons toward the collection optics in the back-scattered Raman geometry. One of the important features of the micro-cavity substrate is that it enhances the entire Raman spectrum of the molecules under investigation and maintains the relative intensity ratios of the various Raman bands. This feature of maintaining the overall integrity of the Raman features during signal enhancement makes the micro-cavity substrate ideal for forensic science applications for chemical detection of residual traces and other applications requiring low sample concentrations. The spectra measured in these cavities are also observed to be highly reproducible and reliable. A simple method for fabricating micro-cavity substrates with precise sizes and shapes is described. It is further shown that micro-cavities coated with nanofilms of gold take advantage of both surface enhanced Raman scattering (SERS) and micro-cavity methods and also significantly improve sample detection limits. PMID- 19281656 TI - Focal length determination of Raman immersion ball probes in diverse media. PMID- 19281657 TI - Ethics, law and moral philosophy of reproductive biomedicine. Foreword. PMID- 19281658 TI - Polar body biopsy: a viable alternative to preimplantation genetic diagnosis and screening. AB - Polar body diagnosis (PBD) is a diagnostic method for the indirect genetic analysis of oocytes. Polar bodies are by-products of the meiotic cell cycle, which have no influence on further embryo development. The biopsy of polar bodies can be accomplished either by zona drilling or laser drilling within a very short time period. However, the paternal contribution to the genetic constitution of the developing embryo cannot be diagnosed by PBD. The major application of PBD is the detection of maternally derived chromosomal aneuploidies and translocations in oocytes. For these indications, PBD may offer a viable alternative to blastomere biopsy as the embryo's integrity remains unaffected, in contrast to preimplantation genetic diagnosis (PGD) by blastomere biopsy. The rapid pace of developments in the field of molecular diagnostics will also influence the advantages of PBD, and probably allow more general diagnostic applications in the future. PMID- 19281659 TI - Embryonic stem cell research and the argument of complicity. AB - While the argument of complicity is only rarely discussed in bioethics, it is of obvious relevance to the issue of imported embryonic stem cells in countries in which the derivation of stem cells from early human embryos is legally prohibited and/or morally rejected. Complicity means that making use of the results or products of an illegal or morally problematic activity is itself morally problematic, although generally to a lesser degree than the original activity. The question arises as to which conditions make the argument of complicity plausible, thus supporting attacks against legislation that aims to promote research based on 'fruits of a forbidden tree'. This paper distinguishes a number of different variants of complicity, proposes that they deserve different kinds of moral valuation and applies the results to the ongoing debate about the German stem cell research law. PMID- 19281660 TI - Stem cells derived from amniotic fluid: new potentials in regenerative medicine. AB - Human amniotic fluid cells have been used as a diagnostic tool for the prenatal diagnosis of fetal genetic anomalies for more than 50 years. Evidence provided in the last 5 years, however, suggests that they can also harbour a therapeutic potential for human diseases, as different populations of fetal-derived stem cells have been isolated from amniotic fluid. Mesenchymal stem cells were the first to be described, which possess the higher proliferation and differentiation plasticity of adult mesenchymal stem cells and are able to differentiate towards mesodermal lineages. Amniotic fluid stem cells have more recently been isolated. They represent a novel class of pluripotent stem cells with intermediate characteristics between embryonic and adult stem cells, as they are able to differentiate into lineages representative of all three germ layers but do not form tumours when injected in vivo. These characteristics, together with the absence of ethical issues concerning their employment, suggest that stem cells present in the amniotic fluid might be promising candidates for tissue engineering and stem cell therapy of several human disorders. PMID- 19281661 TI - Emergency contraception: the state of the art. AB - Emergency contraception, otherwise known as post-coital contraception, is part of the continuum of contraceptive methods that women and couples can use for pregnancy prevention. Although emergency contraception should not be used as a regular, plan-ahead contraceptive method, it gives a woman one last-ditch effort to prevent unwanted pregnancy. This paper reviews the history of emergency contraception, the need for further studies, and results of studies conducted at the World Health Organization. Various methods used for emergency contraception are discussed, as well as their efficacies and side effects. PMID- 19281662 TI - Emergency contraception: how does it work? AB - Emergency (or post-coital) contraception is any substance or device that is used to prevent pregnancy after unprotected intercourse. Currently used hormonal methods of emergency contraception (high-dose combined oral contraceptive pill or levonorgestrel) prevent about 50-80% of pregnancies. Research has demonstrated that these methods inhibit the midcycle surge of LH from the pituitary and, if given at least 2 days before ovulation, ovulation is delayed or prevented. Ovulation still occurs if administration is delayed until ovulation is imminent. Biological data that suggest that the most likely mode of action is by preventing fertilization are supported by the clinical observation that the greater the interval between coitus and administration the greater the chance of pregnancy. There are no data supporting the view that levonorgestrel can impair the development of the embryo or prevent implantation. In contrast, other very effective methods of emergency contraception, such as mifepristone and intrauterine devices, can also inhibit implantation. PMID- 19281663 TI - Is emergency contraception murder? AB - Hormonal emergency contraception (EC) is engendering fierce moral disagreement that is bleeding over into politics and policy. This paper considers Catholic positions on this issue, as they are the fullest and best developed. Its most extreme opponents, such as representatives of the Vatican, hold that EC is an abortifacient that should be banned. Moderates like Sulmasy believe that it should be available to women who have been raped when a negative pregnancy test suggests that fertilization has not yet taken place, and liberals, like Catholics for Free Choice, believe that it should be available to all women regardless of its mode of action. These positions depend in part on underlying philosophical presuppositions about when valuable life begins and scientific assumptions about how EC works. I argue that there are good reasons for rejecting the criterion of fertilization, and that the best current evidence strongly suggests that EC has no post-fertilization effects. These points by themselves undermine key objections to EC. I also show that none of the remaining considerations are sufficiently compelling to warrant overriding women's right to exercise religious, moral, and political agency in preventing undesired pregnancies. PMID- 19281664 TI - Choosing disabilities and enhancements in children: a choice too far? AB - Some parents have taken steps to ensure that they have deaf children, a choice that contrasts with the interest that other parents have in enhancing the traits of their children. Julian Savulescu has argued that, morally speaking, parents have a duty to use assisted reproductive technologies to give their children the best opportunity of the best life. This view extends beyond that which is actually required of parents, which is only that they give children reasonable opportunities to form and act on a conception of a life that is good for them. Does the selection of deaf children violate that responsibility? Morally speaking, parents should refrain from using assisted reproductive treatments or prenatal interventions in order to have a child with a disability. Deafness and other disabilities represent intrinsic disadvantages that cannot be offset by other advantages that families and society can offer to people. By the same token, neither should parents seek enhancements of intelligence or physical traits that would undercut intrinsic goods of human life in similar ways. These moral arguments do not, however, sustain the judgment that the law should necessarily interfere with parents' decisions in these matters, even if those choices are morally unwise. PMID- 19281665 TI - The origins of human sexuality: procreation or recreation? AB - Human sexuality has multiple meanings, although reproduction is considered the focus of sexual activity. In spite of this, human sexuality began to lose its exclusive reproductive meaning very early in the evolution of the genus Homo and, with a concealed ovulation and a female accessible to the male during the entire menstrual cycle, the need became that of avoiding, rather than seeking conception during intercourse. The 'contraceptive revolutions' of the 20th century (sex without reproduction, reproduction without sex, reproduction in menopause and, one day, reproduction without gametes) are having a major impact on the lives of individual couples and women. At the same time, this tumultuous progress drew the attention of religious moralists, since ethics have always focused on sexuality and its moral regulation. Catholic ethicists have been at the forefront of the battle against 'dehumanizing' the reproductive process, whereas Judaism took a much more open position. Early Christian teaching on sexuality, focused on abstinence; this is because Christ himself defined celibacy as a better life choice for human beings. Drawing on this basis, early Church fathers developed the concept, upheld until the 20th century, that intercourse is totally justifiable only in order to procreate. Today, some cautious overtures are being made and the Church has recognized that sexuality can be expression of conjugal love independent from procreation. PMID- 19281666 TI - Ethics and the prohibition of donor gametes in fertility medicine. AB - Some countries and societies ban the use of donor gametes in IVF and other assisted reproductive treatments. Supporters of these bans maintain that third party gametes damage marriage, pose dangers to children and families, put donors at risk, and endanger society at large, but these views are open to moral dispute. In particular, secular moral philosophy does not require couples to rely on their own gametes only to have children. Families can thrive even if children vary in their genetic relationships with parents and siblings. The use of donor gametes merits close attention in matters of safety and oversight, but there is no effect that is so damaging to children, donors or society that justifies closing off their use altogether. Moreover, bans on donor gametes can sometimes be evaded through reproductive tourism, which raises questions of justice for those unable to afford that option. According to one method of measuring moral progress - a method that relies on principles of humaneness and humanity - providing access to donor gametes makes important contributions to happiness and equality. These arguments suggest that the bans against donor gametes in fertility medicine should be reconsidered. PMID- 19281667 TI - What is to be done with surplus embryos? Attitude formation with ambivalence in German fertility patients. AB - Improved pregnancy rates in IVF have led to increasing numbers of surplus embryos which can potentially be used for purposes like donation to another infertile couple or further research. Individuals report high levels of ambivalence concerning the donation of surplus embryos. This study examined which strategies infertile patients use to deal with this ambivalence when asked to evaluate potential dispositions of surplus embryos created during IVF. Guideline-based interviews with fertility patients were audio-recorded and transcribed verbatim. Following the principle of theoretical sampling, eight interviews were analysed by use of Grounded Theory. Analyses focused on processes of individual attitude formation. Strategies for handling ambivalence during attitude formation were identified: the six strategies comprise cognitive and communicative strategies, and were integrated into a model of attitude formation under ambivalence. As ambivalence is a relevant phenomenon in attitude formation within IVF treatment, assessment of ambivalence is strongly recommended in social science studies investigating ethical problems in patient care. In the context of informed consent, there is a need for individual counselling which draws attention to the conflicting values during attitude formation. Counsellors should be aware of the signs of and the strategies to deal with ambivalence. PMID- 19281668 TI - Healthcare innovation and market forces. PMID- 19281669 TI - Contributions from the lifeworld: quality, caring and the general practice nurse. AB - INTRODUCTION: Enhancing quality and safety in primary health systems is of central importance to funders, practitioners, policy makers and consumers. In this paper we explore the roles of general practice nurses in relation to quality and safety. METHOD: Cross-sectional multimethod study of 25 Australian general practices. Using rapid appraisal we collected data for each practice from interviews with practice nurses, general practitioners and practice managers; photographs of nurse-identified 'key workspaces'; structured observation of nurses for two one-hour sessions; and floor plans. RESULTS: Quality was articulated in two domains, reflecting both external and intrinsic determinants. External determinants included a large number of essentially structural, procedural or regulatory processes, the most marked of these being practice accreditation and occupational health and safety; these corresponded to the Habermasian idea of system. Intrinsic determinants related mostly to nurse perception of their own quality behaviour, and consisted of ways and means to improve or optimise patient care; these correspond to Habermas' notion of the lifeworld. DISCUSSION: Nurses describe a productive tension between the regulatory roles that they play in general practices, and patient-focused care, contrary to Habermas' suggestion that system subsumes lifeworld. Current funding systems often fail to recognise the importance of the particular elements of nurse contributions to quality and safety in primary care. PMID- 19281670 TI - Reducing the cost of proton pump inhibitors by adopting best practice. AB - OBJECTIVES: To evaluate the appropriateness of proton pump inhibitor (PPI) use by assessing the level of compliance of PPI prescribing practices with published guidelines and to assess the potential cost avoidance through inappropriate prescribing. METHOD: A six-week observational study of PPI prescriptions was undertaken between April and June 2005, involving hospital inpatients who were taking a PPI prior to admission. The patients were evaluated using a standardised questionnaire to obtain information regarding their PPI use and efficacy. RESULTS: Among the 679 patients admitted during the study period, 133 were receiving a PPI, and of these 97 (50 men and 47 women) were enrolled into the study. The commonest indication for PPI use was gastro-oesophageal reflux disease (GORD, n = 71; 73.2%). In this cohort, more than one-quarter of patients (26.6%) were using greater than the standard PPI dose. Over half of the patients had at least one risk factor known to exacerbate GORD (51.5% were overweight, 46.4% alcohol consumers and 14% current smokers), and 71.1% were receiving medications known to cause or worsen reflux symptoms. Of those patients who reported alarm symptoms, 84% had undergone endoscopy. The overall compliance with the Pharmaceutical Benefits Scheme (PBS) prescribing guidelines was 78.4%, with the major reason for non-compliance being use for non-PBS indications. Estimated cost savings through adoption of recommended prescribing practices and the implementation of step-down therapy for GORD patients were up to AUD 90,866 and AUD 118,456 per 100 patient-treatment-years, respectively. CONCLUSION: PPIs continue to be prescribed outside the treatment guidelines. As a result, opportunities exist to reduce the cost of PPI use through management of contributing factors, adherence to recommended dosage schedules and use of step down therapy in asymptomatic patients where appropriate. PMID- 19281671 TI - Advice to consult a general medical practitioner in Western Australia: could it be cancer? AB - BACKGROUND: Many people will consult a medical practitioner about lower bowel symptoms, and the demand for access to general practitioners (GPs) is growing. We do not know if people recognise the symptoms of lower bowel cancer when advising others about the need to consult a doctor. A structured vignette survey was conducted in Western Australia. METHOD: Participants were recruited from the waiting rooms at five general practices. Respondents were invited to complete self-administered questionnaires containing nine vignettes chosen at random from a pool of 64 based on six clinical variables. Twenty-seven vignettes described high-risk bowel cancer scenarios. Respondents were asked if they would recommend a medical consultation for the case described and whether they believed the scenario was a cancer presentation. Logistic regression was used to estimate the independent effects of each variable on the respondent's judgement. Two-hundred and sixty-eight completed responses were collected over eight weeks. RESULTS: The majority (61%) of respondents were female, aged 40 years and older. A history of rectal bleeding, six weeks of symptoms, and weight loss independently increased the odds of recommending a consultation with a medical practitioner by a factor of 7.64, 4.11 and 1.86, respectively. Most cases that were identified as cancer (75.2%) would not be classified as such on current research evidence. Factors that predict recognition of cancer presentations include rectal bleeding, weight loss and diarrhoea. CONCLUSION: Within the limitation of this study, respondents recommended that most symptomatic people present to their GP. However, we report no evidence that they recognised a cancer presentation, and duration of symptoms was not a significant variable in this regard. Cases that were identified as 'cancer' could not be classified as high risk on the available evidence. PMID- 19281672 TI - What is the importance of the referral letter in the patient journey? A pilot survey in Western Australia. AB - BACKGROUND: Access to specialists is mediated by general practitioners in many countries. In these settings, specialists rely on information in referral letters when deciding which cases to schedule for their clinics. METHOD: Two-hundred and seven consecutive referral letters to gastroenterologists were scored for the amount of information relayed to the specialist, using a published schedule. The 'quality' scores for these referral letters were compared for four groups of patients: patients diagnosed with histological lesion, those with no histological lesion, those who failed to attend clinic, or those who had a diagnosis unknown. Forty-two referral letters were generated with a range of quality scores. Four gastroenterologists were asked to identify which letters described patients 'likely' to have a significant or benign colorectal condition, and whether they could triage the cases for their clinic given only the information in the letters. RESULTS: It was not possible to differentiate which letters related to patients in each of the four categories (P = 0.6). Patients who failed to attend were more symptomatic than those with a histological lesion (35.4 versus 28.2, mean difference 7.14, 95% confidence interval (CI) 14.1 to 0.15, P = 0.045). Patients referred 'urgently' were not, on the basis of the referral letters, the most symptomatic group (29.7 versus 27, mean difference 2.7, 95% CI -3.4 to 8.8, P = 0.38). The specialists failed to agree on the proportion of cases that could be triaged for their clinics. The cases that could be triaged contained more information (mean 66.38 versus 49.86, mean difference 16, 95% CI 1.3-31.7, P < 0.001). CONCLUSION: There was no evidence for an association between the amount of information relayed and the diagnosis of a histological lesion. However, more information was helpful when deciding which patients to schedule first. By corollary, patients referred with lesser documentation of their clinical presentation may be denied 'urgent' access to the gastroenterology clinic. PMID- 19281673 TI - A unifying theory of clinical practice: Relationship, Diagnostics, Management and professionalism (RDM-p). AB - This discussion paper puts forward a unifying theory of clinical practice using three core performance areas of relationship, diagnostics and management, underpinned by professionalism. The model is described, explained and applied to current frameworks for selection, curriculum and assessment. The model is currently being used as a diagnostic tool for defining, exploring and explaining clinically related behaviour in general practice and across a range of hospital specialties, in particular with doctors whose performance has been a cause for concern. There is potential to further apply, develop and research the model in other clinical areas, in non-clinical groups working in health care or other client-based environments. PMID- 19281674 TI - The relevance of equity in health care for primary care: creating and sustaining a 'fair go, for a fair innings'. AB - This paper provides an 'equity lens' through which practitioners, policy makers and researchers can measure and monitor the equity of healthcare services provided. An argument is put forward which shows that services need to be assessed in terms of inequities, as opposed to the more often used terms of inequalities or disparities. This is not just a semantic argument, rather it is based on the foundations that service provision should be socially just. The paper then outlines some of the key domains involved in measuring the equity of health care--access, need and utilisation. The paper concludes with some of the reasons for the current equity problems in healthcare services and suggestions for ways forward in terms of developing more equitable healthcare services. PMID- 19281675 TI - Managing multiple sclerosis in primary care: are we forgetting something? AB - Multiple sclerosis (MS) is the commonest debilitating, progressive neurological disorder in most Western countries. It is important for many reasons, including the personal costs, levels of disability produced, age group affected and the resultant economic burden placed on individuals, families and the community. Although it is thought to be an autoimmune condition, in general little is understood about the causation of MS and the factors that trigger or contribute to exacerbations and deterioration. This overview of the literature will consider some important studies examining the relationship between lifestyle and psychosocial factors and MS progression. These studies suggest that nutrition, sunlight, exercise, stress and social factors can all modulate the rate of progression of MS and the level of disability. Although appearing in respected journals, this information tends to be little known or discussed by clinician and patient alike. If lifestyle approaches do offer potential avenues for therapy, this raises important questions regarding the management of MS in primary care. More widely prescribed conventional medicines have been studied in more detail but are only modestly effective and may have significant side-effects. Are we presently neglecting the most effective approach of combining the non-drug or holistic approach with the best of conventional pharmaceutical therapies, and if so what are the implications of this omission? PMID- 19281676 TI - One service, many voices: enhancing consumer participation in a primary health service for multicultural women. AB - BACKGROUND: Consumer participation in primary health care is important in providing quality consumer-focused care, but challenging when working with disadvantaged groups of diverse cultural and linguistic backgrounds. Women's Health Services (WHS) works with women from over 60 different nationalities, including many newly arrived migrants and refugees. New arrivals access a wide range of WHS programmes including medical services, counselling, information, community talks and workshops, referral, and outreach, but few ethnic women attended the alcohol and other drug (AOD) services offered by the organisation. AIM: To establish an active consumer reference group to assist understanding and reducing the barriers to AOD services for a heterogeneous disadvantaged group that includes individuals from different cultural, language and educational backgrounds. RESULTS: Leaning heavily on experiences from the mental health field, WHS overcame many practical and philosophical considerations which included: agreeing upon the purpose of the group and how it would operate within the structure of the organisation; the level of English language required by participants for the group to function; issues of resourcing the group; and ensuring an appropriate, workable demographic mix in terms of age, language, and migration experiences. CONCLUSION: The process and the outcome of establishing a consumer reference group (CRG) in a primary healthcare setting has been valuable for consumers' and health service providers within the organisation. PMID- 19281677 TI - Learning from four years of collaborative access work in Australia. AB - BACKGROUND: Through the three years of the first phase of the Australian Primary Care Collaborative there was considerable adaptation of the work to improve access translated from the UK to the Australian environment. Changes in four areas are described. METHODS AND RESULTS: MEASURES: 'third available appointment' was retained as a measure of delay. A patient satisfaction survey was revised and a new measure added looking at unmet demand. Team: requests from practices resulted in the production of a set of 'team principles' designed to help practices build capacity for improvement in their teams. Name: the name of the topic seemed to be a barrier for some practices. After much thought, the name of the improvement topic was changed from 'Better Access' to 'Access and Care Redesign'. The product: the content of the access topic was revised. Change ideas were divided into 'foundation work', which all practices were expected to do to improve access to care for patients. Once this was completed, practices were encouraged to select a 'pathway' which best suited their situation. CONCLUSION: Four possible changes are offered for consideration to those planning to do access work with general practices based on the learning from the Australian Primary Care Collaborative. PMID- 19281678 TI - Co-ordination and management of chronic conditions in Europe: the role of primary care--position paper of the European Forum for Primary Care. AB - Healthcare systems in Europe struggle with inadequate co-ordination of care for people with chronic conditions. Moreover, there is a considerable evidence gap in the treatment of chronic conditions, lack of self-management, variation in quality of care, lack of preventive care, increasing costs for chronic care, and inefficient use of resources. In order to overcome these problems, several approaches to improve the management and co-ordination of chronic conditions have been developed in European healthcare systems. These approaches endeavour to improve self-management support for patients, develop clinical information systems and change the organisation of health care. Changes in the delivery system design and the development of decision support systems are less common. Almost as a rule, the link between healthcare services and community resources and policies is missing. Most importantly, the integration between the six components of the chronic care model remains an important challenge for the future. We find that the position of primary care in healthcare systems is an important factor for the development and implementation of new approaches to manage and coordinate chronic conditions. Our analysis supports the notion that countries with a strong primary care system tend to develop more comprehensive models to manage and co-ordinate chronic conditions. PMID- 19281690 TI - Health outcomes in low-income children with current asthma in Canada. AB - Data collected from the Canadian National Longitudinal Survey of Children and Youth (NLSCY) in 1994/95 and 1996/97 were used to measure longitudinal health outcomes among children with asthma. Over 10 000 children aged 1 to 11 years with complete data on asthma status in both years were included. Outcomes included hospitalizations and health services use (HSU). Current asthma was defined as children diagnosed with asthma by a physician and who took prescribed inhalants regularly, had wheezing or an attack in the previous year, or had their activities limited by asthma. Children having asthma significantly increased their odds of hospitalization (OR = 2.52; 95% CI: 1.71, 3.70) and health services use (OR = 3.80; 95% CI: 2.69, 5.37). Low-income adequacy (LIA) in 1994/ 95 significantly predicts hospitalization and HSU in 1996/97 (OR = 2.68; 95% CI: 1.29, 5.59 and OR = 0.67; 95% CI: 0.45, 0.99, respectively). Our results confirmed that both having current asthma and living in low-income families had a significant impact on the health status of children in Canada. Programs seeking to decrease the economic burden of pediatric hospitalizations need to focus on asthma and low-income populations. PMID- 19281691 TI - Deprivation and unintentional injury hospitalization in Quebec children. AB - Injuries disproportionately affect children from deprived areas. This study examines the links between the material and social dimensions of deprivation and injury hospitalizations in children aged 14 years or under from 2000 to 2004. Hospitalization data are from the Quebec hospital administrative data system, whereas socio-economic characteristics of individuals were estimated based on the smallest geographic areas for which Canadian census data were disseminated. The Poisson regression model was used to calculate the relative risks of hospitalization for seven categories of unintentional injury. A total of 24 540 injury hospitalizations were examined. Hospitalization in children is associated with both dimensions of deprivation. Injuries to pedestrians and motor vehicle occupants and injuries related to burns and poisonings are clearly associated with both dimensions of deprivation. These inequalities should be considered in the development of preventive measures. PMID- 19281692 TI - Injury data in British Columbia: policy maker perspectives on knowledge transfer. AB - Provincial and regional decision makers in the injury prevention field were interviewed in British Columbia (B.C.) to obtain their views about best processes for the transfer or dissemination of relevant data. These decision makers (n = 13) indicated that data should provide them with a holistic and comprehensive picture to support their decision processes. In addition, they felt information about injury types and rates should be linked backward to determinants or causes and forward to consequences or outcomes. This complete chain of data is needed for planning and evaluating health promotion interventions. It was also felt that data providers needed to devote more effort to fostering effective receptor capacity, so that injury prevention professionals will be better able to understand, interpret and apply the data. These findings can likely be generalized to other jurisdictions and policy areas, and offer additional insight into the practicalities of knowledge transfer and exchange in researcher/decision maker partnerships. PMID- 19281693 TI - Complementary therapies for cancer patients: assessing information use and needs. AB - Many cancer patients seek complementary therapies (CTs) for cancer management; however, relatively little is known about patients' CT information seeking behaviour. Therefore, we assessed: 1) cancer patients' use of the types and sources of CT information; 2) their information preferences; and 3) their understanding of the phrase "scientific evidence or proof that a therapy works." We collected data from 404 patients attending the Tom Baker Cancer Centre (TBCC) in Calgary and 303 patients calling the Cancer Information Service (CIS) helpline. In most cases, patients wanted information on the safety of CTs, how CTs work and their potential side effects. Physicians and conventional cancer centres were the most desired sources of CT information, but relatively few patients obtained information via these sources. Although patients were aware of the meaning of scientific evidence, they often used information based on non scientific evidence, such as patient testimonials. The creation of a supportive care environment in conventional cancer treatment centres, by providing CT information, may help address cancer patients' concerns and alleviate some of the stress that may have been caused by the cancer diagnosis. PMID- 19281694 TI - Antioxidants for chronic pancreatitis. PMID- 19281695 TI - Spare the steroids, miss the diagnosis? PMID- 19281696 TI - The changing character of acute pancreatitis: epidemiology, etiology, and prognosis. AB - Acute pancreatitis continues to be a diagnostic and therapeutic challenge for physicians and surgeons. It ranks third in the list of hospital discharges for gastro-intestinal diseases. In most patients the cause is either gallstones or alcoholism. The overall mortality is less than 5%, but severe acute pancreatitis leads to prolonged hospitalization and much higher mortality. There are important differences in disease susceptibility and case fatality rates: the incidence is higher in blacks than in whites, and mortality is higher in older patients than in younger patients. Reports from various countries reveal that the frequency of acute pancreatitis is increasing, perhaps in relation to rising obesity rates, which would increase the likelihood of gallstone pancreatitis. Conversely, mortality rates for acute pancreatitis are declining in many, but not all, reports. PMID- 19281697 TI - Prevention, detection, and management of infected necrosis in severe acute pancreatitis. AB - The management of infected peripancreatic or pancreatic necrosis in patients with severe pancreatitis has changed considerably in recent years. This review discusses the recent literature on prevention, detection, and management of infected necrosis. Though antibiotics, probiotics, and enteral nutrition have been tried to prevent infected necrosis, only enteral nutrition has consistently proven to be effective. Antibiotics and probiotics have not shown a consistent beneficial effect on outcome. Enteral nutrition reduced infectious complications and mortality in severe pancreatitis, compared with parenteral nutrition. The detection of infection of pancreatic necrosis is important for clinical decision making. Fine-needle aspiration may be used to confirm suspected infection, but if its results will not change clinical decisions, it should be omitted, as it may even introduce infection. Minimally invasive surgical, radiologic, or endoscopic intervention is increasingly being applied. In the absence of level 1 evidence, local expertise dictates which type of intervention is applied. PMID- 19281698 TI - Endoscopic therapy for chronic pancreatitis: technical success, clinical outcomes, and complications. AB - Chronic pancreatitis (CP) can cause failure of both the exocrine and endocrine portions of the gland. Pain is the most recalcitrant clinical complaint in CP. Generally, conservative measures are first attempted to manage pain. These include cessation of alcohol use and smoking, enzyme replacement therapy, and finally, opioid analgesia. Endoscopy can be employed to treat the pain and complications due to CP. The results of the only two prospective randomized controlled trials suggest that surgery has a more durable effect than endoscopic therapy in controlling pain. Both trials suffer from severe limitations, however, and endoscopy remains the preferred approach for many patients because of its minimally invasive nature. Endoscopic ultrasound celiac plexus block has limited value in helping to control pain. More randomized trials are needed, along with further technologic innovation to improve the current treatment modalities. When considering interventional therapy for a patient with CP, a tailored and multidisciplinary therapeutic approach should be taken. PMID- 19281699 TI - Neuroendocrine tumors of the pancreas. AB - Pancreatic endocrine tumors are rare neoplasms accounting for less than 5% of pancreatic malignancies. They are broadly classified into either functioning tumors (insulinomas, gastrinomas, glucagonomas, VIPomas, and somatostatinomas) or nonfunctioning tumors. The diagnosis of these tumors is difficult and requires a careful history and examination combined with laboratory tests and radiologic imaging. Signs and symptoms are usually related to hormone hypersecretion in the case of functioning tumors and to tumor size or metastases with nonfunctioning tumors. Surgical resection remains the treatment of choice even in the face of metastatic disease. Further development of novel diagnostic and treatment modalities offers potential to greatly improve quality of life and prolong disease-free survival for patients with pancreatic endocrine tumors. PMID- 19281700 TI - Laparoscopic pancreatic surgery: what now and what next? AB - In recent years, improved laparoscopic skill sets have expanded surgical management of pancreatic disease to encompass pancreatic resection, tumor enucleation, debridement, and drainage. With the aid of radiologically guided drainage catheters, necrosectomy for acute pancreatitis can be delayed and accomplished laparoscopically in a select patient population. Pancreatic pseudocysts from chronic pancreatitis can now be approached via minimally invasive strategies, including emerging combined laparoscopic procedures and natural orifice transluminal endoscopic surgery. It is clear that laparoscopic pancreaticoduodenectomy is possible in experienced hands; pancreatic neoplasms in the body and tail are more suitable for laparoscopic procedures because distal pancreatic resection requires no reconstruction of the biliary or enteric tract. Laparoscopic staging of pancreatic tumors has decreased as preoperative radiographic imaging becomes more sensitive. Similarly, laparoscopic palliative procedures have decreased because of the emergence of other minimally invasive options for relieving gastric outlet obstruction and biliary obstruction. Nonetheless, major advances in minimally invasive pancreatic surgery will continue as technology and skill sets advance. PMID- 19281701 TI - Minimizing complications in pancreaticobiliary endoscopy. AB - Endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic ultrasound are invaluable tools in the diagnostic and therapeutic evaluation and management of a variety of pancreatobiliary disorders. Along with a significant refinement in the equipment and techniques used has come a recent trend toward aggressive therapeutic interventions. Because of the technical nature of these procedures and the characteristics of the patients, post-procedural complications may occur, ranging from minor (requiring brief hospitalization) to severe (causing permanent disability or death). This review summarizes these complications and outlines strategies to minimize them. PMID- 19281702 TI - Preoperative biliary drainage in malignant obstruction: indications, techniques, and the debate over risk. AB - In patients with malignant biliary obstruction considered suitable for surgical resection, preoperative drainage might be expected to improve surgical outcomes by restoring liver function and improving nutritional status. In practice, however, the benefits of preoperative drainage are far from clear. Studies to date have reported differing outcomes, and some have suggested that morbidity and mortality are greater in patients undergoing drainage than in those proceeding directly to surgery. The development of clinical guidelines has been limited by the lack of convincing randomized data, which in turn has led to variations in practice. This article examines the arguments for and against preoperative biliary drainage, the conflicting data on the subject, and the techniques used. PMID- 19281703 TI - Endoscopic ultrasound in the evaluation of radiologic abnormalities of the liver and biliary tree. AB - Gastroenterologists increasingly find themselves in the un-enviable position of having to choose the optimal radiographic test to visualize the biliary tree and the liver. This dilemma is compounded by the rapid evolution of the available technologies and their ever-increasing resolution capabilities. Endoscopic ultrasound (EUS) has shown itself to be equally capable of providing detailed imaging of the biliary tree and of surrounding structures; its potential for evaluating liver lesions is just now being realized. EUS-fine needle aspiration (FNA) may supplant some of the existing technologies as it offers a way to sample masses, strictures, lymph nodes, and now certain segments of the liver and periportal regions. This review takes an in-depth look at the role of EUS in evaluating bile duct dilatation, suspected choledocholithiasis, biliary strictures, and polyps and masses of the gallbladder. It also reviews the literature on the exciting and rapidly evolving role of EUS-FNA evaluation of occult, metastatic, and primary hepato-cellular malignancies, as well as periportal lesions. Technologies such as EUS elastography may completely obviate FNA of liver lesions. PMID- 19281704 TI - The investigation of unexplained biliary dilatation. AB - The investigation of biliary dilatation forms a routine part of gastroenterology practice. In developed countries, biliary dilatation is usually the result of obstruction of bile flow by either stones or mitotic lesions of the pancreas or biliary tree, and standard radiologic and endoscopic techniques are used to identify and relieve the obstruction. In the absence of an obvious cause, however, the investigation and management of biliary dilatation can prove challenging, particularly while trying to minimize invasive studies. This review examines factors thought to influence bile duct size in the absence of obvious obstructing pathology and looks at some causes of biliary dilatation that are unusual and potentially difficult to diagnose. PMID- 19281705 TI - ERCP in the management of biliary complications after cholecystectomy. AB - Open cholecystectomy has been associated historically with 0.2% to 0.5% risk of postoperative biliary injury. Laparoscopic cholecystectomy, which has become the first-line surgical treatment of calculous gallbladder disease, has been associated with a 2.5-fold to fourfold increase in the incidence of postoperative bile duct injury. The biliary endoscopist can expect to see a varied spectrum of complications after cholecystectomy by either technique, including postoperative biliary strictures, bile leaks, and retained calculi in the biliary tree. Proper diagnosis and treatment are paramount in ensuring a satisfactory outcome after bile duct injury. Endoscopic retrograde cholangiopancreatography (ERCP) has become the primary modality for treatment and effectively manages most bile duct injuries. PMID- 19281706 TI - Epidermodysplasia verruciform-like Lesions in an HIV patient. AB - Epidermodysplasia verruciformis (EV) is a rare disorder involving widespread infection with specific human papillomavirus types and characteristic clinical lesions that may resemble verruca plana, tinea versicolor, psoriasis, or seborrheic keratoses. The most common HPV types found in EV are 5, 8, 17, and 20. Histopathologically, lesions demonstrate stereotypical enlarged keratinocytes in the upper epidermis with gray-blue cytoplasm, enlarged round nuclei with pale chromatin, and one or multiple nucleoli. Epidermodysplasia verruciformis may occur in either a classical form (often familial, early onset, and complicated by squamous cell carcinoma) or in association with various hereditary or acquired immunodeficiencies, particularly HIV. Fewer than 20 cases of HIV-associated epidermodysplasia verruciformis have been reported. We describe a 42-year-old HIV positive man who presented with hypo- and hyperpigmented papules and plaques on the upper trunk, head, and neck, with histopathologic findings of epidermodysplasia verruciformis. PMID- 19281707 TI - Multicentric reticulohistiocytosis in a Malaysian Chinese lady: a case report and review of literature . AB - A 36-year-old Malaysian woman with erosive arthritis presented with multiple asymptomatic skin colored papulonodules on the hands, elbows, knees, ears, and feet; many macules, papules, and plaques were photodistributed. Progressive, erosive arthritis began about 18 months after the onset of skin involvement. Initially the patient was diagnosed to have dermatomyositis due to her skin findings, but later proved to have multicentric reticulohistiocytosis based on the typical histopathological features of histiocytic multinucleated giant cells with eosinophilic ground glass cytoplasm on skin biopsy. Workup for associated diseases including malignancies was negative. Her skin lesions and joint pain responded to treatment with methotrexate and prednisolone. PMID- 19281708 TI - Fluoroscopy-induced chronic radiation dermatitis: a report of three cases. AB - We report these cases to emphasize the importance of recognizing fluoroscopy as a cause of radiation dermatitis. The diagnosis of fluoroscopy-induced chronic radiation dermatitis should be raised when patients present with morpheaform, telangiectatic, or ulcerative skin findings in the characteristic locations. PMID- 19281709 TI - A sticktight flea removed from the cheek of a two-year-old boy from Los Angeles. AB - Species-level identification of an ectoparasite often requires outside expertise. Patients are increasingly turning to the internet for medical information, in part to gain access to experts. We present a case of a flea identified after contact was made between a patient's father and the authors via an insect-related forum on the internet. The flea had been attached for 12 hours without inducing symptoms before it was removed. The specimen was identified as a sticktight flea Ectoparasites in the differential diagnosis for this case are discussed. PMID- 19281710 TI - Cheilitis glandularis and actinic cheilitis: differential diagnoses - report of three unusual cases. AB - A variety of pathological conditions of diverse etiologies may involve the lips. Cheilitis glandularis (CGL) is a rare disorder of unknown etiology characterized by inflammation of the minor salivary glands of the lower lip. The aim of this paper is to compare the diagnosis and treatment of two cases of CGL at an early age (one in a child and the other in a young adult with mental retardation) with the diagnosis and treatment of actinic cheilitis, which is much more prevalent than CGL in tropical countries. PMID- 19281711 TI - Extragenital bullous lichen sclerosus atrophicus. AB - Lichen sclerosus et atrophicus (LSA) is a rare, chronic, mucocutaneous disease of unknown etiology. Onset can occur in subjects of any age, but the condition is more prevalent in adult females around the time of menopause. In both sexes, the most common site of the lesions is the anogenital area. Bullous lichen sclerosus et atrophicus is an unusual form of the disease and the exact prevalence is not known. We describe a case of nongenital bullous LSA in a man. PMID- 19281712 TI - Pseudoxanthoma elasticum. AB - A 55-year-old man with progressive loss of vision was referred for dermatology consultation for the evaluation of his skin lesions. The cutaneous examination of the patient revealed multiple small yellow papules, coalescing into plaques, on his neck, axillae and periumbilical regions. He also had redundant skin folds on the axillae. His peripheral pulses and blood pressure were normal. Angioid streaks were found in the ocular fundi. A skin biopsy specimen of the papule showed fragmentation of elastic fibers as well as calcification in the dermis. Given the clinical manifestations and the histopathologic findings, the patient's illness was diagnosed as pseudoxanthoma elasticum. The patient was then sent to undergo a thorough cardiovascular evaluation. PMID- 19281713 TI - A case of acquired digital fibrokeratoma accompanied by pyogenic granuloma. AB - A 35-year-old male presented with a slowly growing keratotic tumor on the plantar aspect of his left second toe. Over a ten-year period, the projecting tumor had grown to the size of 7 x 18 mm without causing any symptoms. Three months prior to presentation, a red and polypoid papule of 5 x 5 mm-size began to develop within the original tumor. The tumor was diagnosed as acquired digital fibrokeratoma accompanied by pyogenic granoloma. Hereby, we report this unusual case. PMID- 19281714 TI - White fibrous papulosis of the neck: the first South American case. AB - A 69-year-old woman presented with white papules on her neck. These papules were multiple, white, discrete, asymptomatic, non-follicular, 2 to 3 mm in diameter, and located on both sides of the neck. On histopathological examination, there was a thickened and abnormal organization of collagen bundles in the papillary to mid-dermis and a decrease in the number of elastic fibers. This clinical and histological picture suggested the diagnosis of white fibrous papulosis of the neck. No treatment was given because of the benign nature of the disease. This is the first case report of this condition in South America. PMID- 19281715 TI - Pairing and comparing nine diseases with Degos Disease (Malignant Atrophic Papulosis): an attempt to illustrate our understanding and direct future inquiry. AB - Degos disease is a poorly defined condition that encompasses cutaneous and systemic findings that overlap with a large number of other rheumatologic and coagulation disorders. Comparisons are made herein with 9 conditions that share findings and presentations. PMID- 19281716 TI - Keratolytics for psoriasis: are they necessary? AB - It is a common belief that keratolytic agents are required to enhance the penetration of topical medications into thick psoriatic plaques. However, is this belief evidence-based? PMID- 19281717 TI - Parable: the broken reed. PMID- 19281718 TI - A neglected issue in interpretation of results of randomized controlled trials: informative censoring. AB - The presence of informative censoring is a commonly neglected issue that can lead to inaccurate results in randomized controlled trials. It is mandatory to perform an intention to treat analysis to minimize this source of error. PMID- 19281719 TI - Comparison of podophyllotoxin and imiquimod as anal condyloma acuminata therapy. AB - Herein we revisit the published evidence for comparison of the efficacy of imiquimod and podophyllotoxin for anal condyloma acuminata. PMID- 19281720 TI - Allergic contact dermatitis to pure henna. AB - Henna is a naturally occurring brown dye made from the leaves of the tree Lawsonia inermis. The active ingredient of henna is lawsone (2-hydroxy-1, 4 naphthoquinone). It is traditionally used in Islamic and Hindu cultures as a hair coloring and as a dye for decorating the nails or making temporary skin tattoos. Actually, henna has a very low allergic potential. In most cases, allergic reactions not caused by henna, but by the chemical coloring additives that are added to henna mixtures. These additives include agents such as daiminotoluenes and diaminobenzenes. In this article, we report a case of allergic contact dermatitis from pure henna that is also used for the relief of rheumatic pain. PMID- 19281721 TI - Risk factors of recurrent furunculosis. AB - This is a case control study of recurrent versus non-recurrent furunculosis. The aim was to identify the risk factors and causative micro-organisms associated with recurrence of furunculosis. Data was collected through an anonymous questionnaire. Weight and height were measured to calculate body mass index and a specimen was drawn from a lesion for culture. A nasal swab was taken to identify nasal Staphylococcus aureus carriers. Staphylococcus aureus was the causative organism in 89 percent and 100 percent of recurrent and non-recurrent furuncles, respectively. No significant difference was detected in resistance to the commonly used antibiotics. Logistic regression analysis revealed that the most important independent predictor of recurrence was a positive family history. The other independent predictors were anemia, previous antibiotic therapy, diabetes mellitus, previous hospitalization, multiplicity of lesions, personal hygiene and associated skin diseases. Nearly all these risk factors are modifiable, preventable, and/or controllable by simple measures such as sound personal hygiene and a balanced diet. Further research is needed to study if the same organism is implicated in recurrent cases and to test the feasibility as well as cost-benefit of prophylactic use of antibiotics. It is not advisable to use antibiotics to clear nasal carrier of S. aureus in our culture with the large number of possible sources of re-infection other than nasal nares. PMID- 19281722 TI - Pharmacotherapy guidelines for the aged by family doctors for the use of family doctors: part C--Special pharmacology. AB - The part "Special pharmacology of the aged" of this guideline contains recommendations for typical conditions in the family doctors practice: in the January issue 2009 dementia and Morbus Parkinson, in this issue osteoporosis and urinary incontinence and in the next issue rectal incontinence and obstipation. This issue of the IJCPT contains the third part of the Pharmacotherapy guidelines for the aged by family doctors for family doctors. Part 3: Osteoporosis and urinary incontinence. Osteoporosis is a systematic disease characterized by low bone mass and declining bone structure. Exercise, adequate diet, nicotine abstinence as well as reduction of alcohol consumption may counteract the progression of the disease. Osteoporosis manifests in bone fractures with minimal trauma. Attention must be given to the risk of falling, e.g., by avoiding drugs that increase the risk of falling: e.g., psychotropic agents, analgesic drugs and antiarrhythmic agents. Specific osteoporosis medication e.g. calcium, vitamin D, biphosphonates and SERM (selective estrogen receptor modulators) is evaluated by family doctors according to indication, dosage, contraindications, long-term therapy and nature of any fracture. Duration of therapy is at least 3 - max. 5 years followed by reassessment of indication. There are 3 types of urine incontinence (urge-, stress-, and overflow-incontinence). Another standardization of urinary incontinence follows dysfunctions of the pelvic floor: detrusor muscle dependent, due to sphincter spasm, prostate gland dependent. Urge incontinence with a dysfunction of the detrusor muscle is the most common type. Mixed types are frequent. Non-drug measures (e.g. pelvic muscle training, bladder training, toilet training are first choice treatments. Drug therapy (estrogen, imipramine) are without proven effect. PMID- 19281723 TI - Accuracy assessments of quantitative diagnostic tests for clinical research. AB - BACKGROUND: Clinical research is impossible without accurate diagnostic tests. The methods for assessing accuracy of quantitative diagnostic tests are not routinely used by the scientific community. OBJECTIVE AND METHODS: To review the advantages and disadvantages of methods that could be used for that purpose. Using real data examples we review seven possible methods. RESULTS AND CONCLUSIONS: Simple linear regression testing the presence of a significant correlation between the new test data (x-axis data) and the control test data (y axis data) is not accurate for testing the validity of a novel quantitative diagnostic test. Accurate methods using linear regression include the following. First, from y = a + b x, test the hypothesis that b is statistically significantly larger than zero, than test the hypothesis that b = 1.000 and a = 0.000. Second, if "the b = 1.000 and a = 0.000 hypothesis" cannot be confirmed, then use as criterion for validation a squared correlation-coefficient r2 or intraclass correlation of > 95%, or a relative residual variance of < 5%. If the new test is validated this way, then the predicted control-test-values are calculated from the equation y = a + bx. The above three methods assume uncertainty of the new test data, but not of the control test data. Deming regression, Passing-Bablok regression, paired Student's t-tests, and Altman-Bland plots assume uncertainty of both the new test and the control test. This is rarely a condition for validation, and carries the risk of unneeded loss of sensitivity of testing. However, if the control test is not the gold standard test and it is decided to account the uncertainty of the control test, then Passing-Bablok regression is the only method that adjusts for non-normal data as frequently observed in practice. PMID- 19281724 TI - Drug exposure and perceived adverse drug events reported by liver-transplant patients. AB - INTRODUCTION: Posttransplant patient outcome and quality of life are affected by different factors, such as post-graft context, psychological state, and polymedication. Many surveys have been carried out to study immunosuppressant ADRs, and have mainly used a questionnaire completed by patients, but few have asked patients about their drug exposure. The aim of this study is to describe drug exposure and adverse drug events (ADEs) reported by liver-transplant patients (LTP). METHODS: This observational, retrospective study assessed questionnaires from LTPs concerning demographic data, drug exposure, and ADEs. RESULTS: 118 LTPs exposed to 5.9 (+/- 2.8) drugs with immunosuppressive regimens, consisting mainly of tacrolimus (79.3%), cyclosporine (18.1%), or sirolimus (2.6%), were also exposed to antihypertensive drugs (43.2%), protonpump inhibitors (30.5%), statins (28.8%), drugs acting on bile composition (26.3%), and diuretics (19.5%). 1,389 ADEs were reported: 30.1% neurological, 13.4% cutaneous, 12.4% hematological, 11.1% digestive, 10.1% osteomuscular, 6.6% cardiovascular, and 16.3% others. Significantly more ADEs were reported by patients exposed to cyclosporine than those receiving tacrolimus (p < 0.05). Patients with a transplant for < 18 months had more tremors and those with a transplant for > 79 months reported more hirsutism, gingival hypertrophia, and arterial hypertension. CONCLUSIONS: This study shows the value of patient reporting via structured interviews for both drug exposure and ADEs, and the importance of this approach to complement total data collection. PMID- 19281725 TI - Ciprofloxacin-related acute severe myalgia necessitating emergency care treatment: a case report and review of the literature. AB - OBJECTIVE: To report a case of an uncommon and up to date unpublished peracute and overwhelming muscle pain following administration of ciprofloxacin. CASE SUMMARY: A 58-year-old male developed fulminating musculoskeletal pain associated with third-time exposure to ciprofloxacin administered for treatment of chronic otitis media. Symptoms resolved slowly after intense combined analgetic therapy and cessation of ciprofloxacin therapy. 24 h after ciprofloxacin discontinuation the symptoms had completely disappeared and no more analgetic treatment was needed. Laboratory values, especially muscle enzymes, did not reveal any pathological pattern. The patient's past medical history highlighted reproducible side effects with both systemic and local administration of ciprofloxacin including milder symptoms of the musculoskeletal system. DISCUSSION: Common side effects of fluoroquinolones include gastrointestinal, central nervous and allergic reactions, but also more uncommon reactions such as tendonitis and rhabdomyolysis. In our case, there had been no signs of rhabdomyolysis. Besides an elevated IgE level no clinical signs of a true anaphylactic reaction associated with release of mast cell mediators had been observed. A pharmacokinetic interaction between ciprofloxacin and the patient's comedication carbamazepine is unlikely to be the responsible mechanism, since fluoroquinolones inhibit cytochrome P450 isoenzyme CYP1A2 but not CYP3A4 which metabolizes carbamazepine. CONCLUSION: To our knowledge, this is the first report describing fulminating musculoskeletal pain following administration of ciprofloxacin without any signs of rhabdomyolysis. Physicians should notice that there is a variety of adverse reactions of this usually well-tolerated agent and they should be aware of unusual complaints of their patients who receive fluoroquinolone treatment. PMID- 19281726 TI - In vitro and in vivo pharmacokinetic characteristics of clazosentan, an intravenous endothelin receptor antagonist, in humans. AB - OBJECTIVE: In this study, the distribution, metabolism and excretion of the endothelin receptor antagonist clazosentan were investigated. SUBJECTS AND METHODS: 4 healthy male subjects received an intravenous 3-h infusion at a rate of 0.2 mg/kg/h of 14C-labeled clazosentan and blood, urine and feces samples were collected for a period of 8 days. Experiments were performed to investigate the plasma protein binding, the binding to red blood cells and the inhibition potential of cytochrome P450 isoenzymes of clazosentan. RESULTS: Clazosentan was mainly excreted unchanged into feces whereas about 15% of the radioactive dose was recovered in urine. No metabolites representing more than 5% of total radioactivity were identified. No relevant inhibition of the human cytochrome P450 isoenzymes, 1A2, 2A6, 2B6, 2C9, 2C19, 2D6, 2E1 and 3A4, was observed in vitro at clazosentan concentrations largely exceeding those observed in clinical trials. In human blood, clazosentan was highly bound to plasma proteins and did hardly penetrate into red blood cells. CONCLUSION: The primary route of excretion of clazosentan was via the feces, mainly as unchanged drug. PMID- 19281727 TI - Daptomycin elimination by CVVH in vitro: evaluation of factors influencing sieving and membrane adsorption. AB - OBJECTIVE: Knowledge on the elimination of antibiotics by extracorporeal hemofiltration is a prerequisite for appropriate antimicrobial dosing in patients with renal failure. The present study set out to determine the clearance of the novel lipopetide antibiotic daptomycin from human whole blood by continuous venovenous hemofiltration (CVVH) in vitro. In addition, factors influencing daptomycin sieving and membrane adsorption were investigated. METHODS: A recirculation model using different solvent media was established and daptomycin was added to the simulated blood circuit at varying concentrations. The concentration of daptomycin over time in the modelled blood compartment and the ultrafiltrate was measured by high performance liquid chromatography (HPLC). RESULTS: Mean Sieving coefficients (SCs) of daptomycin over time were calculated to 0.98 +/- 0.05, 0.33 +/- 0.02 and 0.40 +/- 0.03 at a baseline concentration of 60 microg/ml in Ringer lactate, Ringer lactate containing human albumin and in human whole blood, respectively. SCs of daptomycin in protein-containing media were higher than the free fraction in plasma of approximately 10%. Neither concentration of daptomycin nor addition of a second antibiotic showed significant impact on the SC. Adsorption of daptomycin to synthetic surfaces proved moderate and saturable, resulting in loss of around 20% of the amount initially added to the artificial blood circuit. CONCLUSION: In our in vitro setting the calculated clearance of daptomycin from whole blood exceeded the physiological clearance described for individuals with normal renal function. Investigation of clearance by CVVH in vivo seems necessary. Until sufficient clinical data are available for patients undergoing CVVH, monitoring of daptomycin concentrations in this population might be recommended in order to avoid sub-therapeutic exposure to daptomycin. PMID- 19281728 TI - Single-dose randomized, open-label, 2-way crossover bioequivalence study of clopidogrel 75 mg tablet in healthy volunteers under fasting conditions. AB - AIM: This study aimed to assess the bioequivalence of 2 formulations of 75 mg clopidogrel hydrogen sulphate film-coated tablet, under fasting conditions. SUBJECTS AND METHODS: 64 healthy subjects, age ranging from 19 to 55 years, were enrolled in a single-centre, randomized, single-dose, open-label, 2-way crossover study, with a minimum washout period of 7 days. Plasma samples were collected up to 24 h post dosing. Clopidogrel and clopidogrel carboxylic acid levels were determined by reverse-phase high-performance chromatography coupled to tandem mass spectrometry detection, LC-MS-MS method. Pharmacokinetic parameters used for bioequivalence assessment were the AUClast (area under the concentration-time curve from time zero to time of last observed nonzero concentration) and the Cmax (maximum observed concentration). These parameters were determined from the clopidogrel concentration data using non-compartmental analysis as well for clopidogrel carboxylic acid concentration data. RESULTS: The 90% CI (90% confidence intervals), obtained by analysis of variance (ANOVA) were within the predefined ranges (80.00 - 125.00%) for both analytes. CONCLUSION: Bioequivalence between test and formulations, under fasting conditions, was concluded both in terms of rate and extent of absorption. PMID- 19281729 TI - Bioequivalence study of a new oral topotecan formulation, relative to the current topotecan formulation, in patients with advanced solid tumors. AB - OBJECTIVE: The aims of this study were to investigate the bioequivalence of a new oral topotecan formulation (i.e., proposed commercial formulation) relative to the current oral formulation (formulation used in previous clinical trials), the effect of food on the absorption and disposition of the new oral topotecan and its safety and tolerability in patients with advanced solid tumors. PATIENTS AND METHODS: This was a multi-center, pharmacological Phase I, multiple-dose, randomized, open-label, cross-over bioequivalence study. In the bioequivalence part, 85 patients were randomized to receive either a 4 mg (4 x 1 mg) dose of the new or current formulation on Days 1 or 8. In the food-effect part, 23 patients received a 4 mg (4 x 1 mg) dose of the new formulation in a fasted and fed state. Total topotecan and topotecan lactone were determined and pharmacokinetic data were analyzed by non-compartmental method. RESULTS: Bioequivalence was demonstrated as the 90% confidence intervals of the ratio of the new to current formulation for both the area under the plasma concentration-time curve (AUC) and the maximal drug concentration (Cmax) for topotecan lactone were contained within the 0.8 - 1.25 boundary. The AUC and Cmax were similar in the fed and fasted state whilst food delayed the tmax for topotecan lactone and total topotecan. Safety data were collected on all subjects enrolled (n = 108) and were consistent with observations from previous studies of oral topotecan. All subjects experienced at least one adverse event, the majority of which were graded as mild to moderate in severity. CONCLUSION: The new oral topotecan formulation demonstrated bioequivalence to the current formulation and demonstrated it can be administered to patients with solid tumors in the fed or fasted state with similar systemic exposure. PMID- 19281730 TI - Comparison of bioavailability of two ubidecarenone products in healthy Korean volunteers. AB - OBJECTIVE: This study aimed to evaluate the bioavailability of two pharmaceutical products of ubidecarenone (coenzyme Q10, CoQ10). MATERIALS: Two brands (brand A and brand B) of commercial CoQ10 hard capsules. METHODS: Two brands of CoQ10 capsules were administered at 100 mg dose to two groups of healthy volunteers, respectively, and blood samples were withdrawn at predetermined time intervals and assayed by a validated HPLC method with an electrochemical detector. RESULTS AND CONCLUSIONS: Intra- and inter-day precision and inter-day accuracy were acceptable for all quality control samples including the lower limit of quantitation of 50 ng/ml. Recovery of CoQ10 from human plasma was greater than 98.2%. CoQ10 was stable in human plasma under various storage conditions. This method was applied to a pharmacokinetic study after oral administration of CoQ10 hard capsules to healthy volunteers. The intrinsic CoQ10 concentrations were measured for three consecutive days before drug administration, which were ranged between 0.68 and 0.79 microg/ml, and there was no statistically significant difference between groups. In brand A, the plasma concentration after administration of CoQ10 was not higher than the intrinsic level, indicating that no significant drug absorption occurred, whereas considerably higher concentrations were obtained with brand B. The dissolution rates of brand A and B after 3 h were 0.35 +/- 0.09 and 1.27 +/- 0.16%, respectively. From the adjusted concentration-time curve, the AUC and t1/2 of brand B were calculated to be 11.51 +/- 5.76 microg x h/ml and 21.7 h, respectively. A mean Cmax of 0.32 +/- 0.1 microg/ml was obtained at 7.9 h. In conclusion, it was found that bioavailability of CoQ10 was significantly different depending on the formulations, and dissolution could be one of the important factors affecting CoQ10 absorption. PMID- 19281731 TI - Relative bioavailability and pharmacokinetic study of omeprazole 20 mg enteric coated tablet in healthy Bangladeshi volunteers. AB - OBJECTIVE: Introduction of omeprazole constituted a break through in the management of acid-related gastric disorders. Omeprazole effectively suppresses the gastric acid secretion in the parietal cells of the stomach. It is a widely prescribed proton pump inhibitor in Bangladesh. The increasing number of omeprazole containing products available in the market raises questions of therapeutic equivalence and/or generic substitution which are yet to be conducted on the Bangladeshi population. The aim of the study is to assess the relative bioavailability and pharmacokinetic properties of two oral formulations of 20 mg omeprazole tablet, namely LOSEC(R) as reference product and Losectil DR as test product using serum data. MATERIALS AND METHODS: The randomized, two-way crossover study was conducted on 24 healthy male subjects in compliance with the Declaration of Helsinki and ICH Guidelines. Subjects were assigned to receive Losectil DR (Test) and LOSEC (Reference) as a single dose of 20 mg tablet under fasting conditions, following a washout period of 1 week. After oral administration, blood samples were collected at various time intervals and analyzed for omeprazole concentrations using a validated HPLC method. The pharmacokinetic parameters were determined by a non-compartmental method. RESULTS: From serum data, the obtained values for test and reference products were 593.05 +/- 84.85 and 607.92 +/- 67.07 ng/ ml for Cmax; 1756.71 +/- 287.29 and 1786.90 +/- 280.17 ng-h/ml for AUC0-24; 1889.26 +/- 286.46 and 1929.18 +/- 284.33 ng-h/ml for AUC0- yen, respectively. No statistically significant differences were observed between two formulations by analyzing different pharmacokinetic parameters in terms of period, sequence and formulation. From the paired t-test, no significant differences between two formulations were observed (p > 0.05). The 90% CIs of Cmax, AUC0-24 and AUC0- yen were found to be 91.59 - 122.60%, 101.86 - 116.78% and 102.77 - 116.68%, respectively, which are within the FDA accepted limits for bioequivalence (80 - 125%). CONCLUSION: Finally it can be concluded that both products are bioequivalent in terms of rate and extent of drug absorption and therefore interchangeable. PMID- 19281733 TI - The message for World Kidney Day 2009: hypertension and kidney disease--a marriage that should be prevented. PMID- 19281734 TI - Prevalence of chronic kidney disease: a systematic review and meta-analysis. AB - AIMS: Many studies have estimated the prevalence of chronic kidney disease (CKD) but results have varied due partly to the type of equation used to estimate GFR, type of subjects, and ethnicity. This review aimed to estimate the prevalence of CKD Stage III, accounting for these factors. METHODS: 403 studies were identified from Medline using the PubMed search engine, of which 34 studies were eligible. Data were independently extracted by two reviewers, and heterogeneity was assessed using metaregression. RESULTS: The pooled prevalence was estimated using a random effects model. In the general population, the prevalences of CKD Stage III using MDRD equation were 3.6% (95% CI: 2.5, 4.8), 10.7% (95% CI: 4.5 - 16.9%), and 16.3% (95% CI: 2.1 - 30.5%) for age groups 60 years. The prevalence was about double using the Cockcroft-Gault equations, i.e. 7.5% (95% CI: 6.9 - 8.2%) and 34.9 (95% CI: 25.9 - 44.8%) in age 50 years, respectively. The prevalence was similar in Caucasians and Asians aged. < or = 60, i.e. 9.9 versus 9.3%. The prevalence was also higher in the diabetic population than in the general population, i.e. 18.2% versus 10.6%. CONCLUSIONS: The pooled prevalence of CKD in the general population varied according to age groups. The prevalence is similar in Caucasians and Asians within age 60 years or younger but other age groups need more studies in order to pool. Individual patient meta-analysis would be appropriate to resolve the causes of heterogeneity. PMID- 19281735 TI - Metabolic syndrome attenuates effect of chronic kidney disease on prevalence of coronary disease in men referred for stress imaging study. AB - BACKGROUND: Metabolic syndrome (MS) and chronic kidney disease (CKD) are both strongly associated with coronary artery disease (CAD). Components of MS also cause CKD. The incremental effect of CKD on CAD prevalence in MS patients referred for stress imaging studies is unknown. METHODS: From January to December 2005, consecutive subjects referred for a stress imaging study were prospectively enrolled. CAD was defined as fixed or reversible defects on nuclear imaging and as resting or stress-induced wall motion abnormalities on echocardiography. MS was defined using NCEP-ATP III criteria. CKD definition was based on calculated glomerular filtration rate. The independent effect of CKD on stress results was assessed using multiple variable logistic regression. Stepwise model selection was used for variable reduction, and areas under the receiver operating characteristic curves (ROC) were calculated. RESULTS: Of 1,122 patients enrolled (mean age 61.4 years, 97% male), 535 (47%) had MS. Among MS patients, 156/535 (29%) had CKD while 116/587 (19%) subjects without MS had CKD. Subjects with CKD were older (p < 0.001) in subjects with and without MS. The presence of CKD affected prevalence of CAD in the non-MS group only, almost doubling it (20% vs. 38%, p < 0.001). Further, using the ordered nature of the 5 CKD stages, worsening severity of CKD had greater prevalence of CAD, in non-MS subjects only (p < 0.001). CONCLUSIONS: MS attenuates the effect of CKD on CAD prevalence, regardless of CKD severity. CKD almost doubles the prevalence of CAD in non-MS subjects. CKD severity is associated with greater CAD burden in the non-MS group. PMID- 19281736 TI - Prevalence of heparin-induced antibody in African-American hemodialysis patients- comparison to non-dialysis patients. AB - AIMS: Heparin-induced antibody formation and thrombocytopenia are well established complications of heparin use and may be associated with thromboembolic events. Our study aimed to establish the prevalence of heparin induced antibodies (HIA), otherwise known as platelet factor 4 (PF4) antibodies, and their relationship to thrombocytopenia and thrombotic events in a group of predominantly African-American hemodialysis patients over 12 months. For comparison, the prevalence of HIA in thrombocytopenic patients with serum creatinine of less than 2.0 mg/dl tested for the antibody in the same period was evaluated. METHODS: 36 hemodialysis patients on thrice-weekly, in-center treatments receiving heparin on dialysis were selected to have blood drawn predialysis for PF4 heparin antibodies (HIA). Complete blood counts (CBC) were also checked that day and monthly for the next 3 months. Outpatient records were reviewed for thromboembolic events, including thrombosis of dialysis access lines, grafts and fistulas. The HIA results for the dialysis patients were compared with HIA performed at the same lab for non-dialysis thrombocytopenic patients during the same period. RESULTS: 17 out of 36 (47%) hemodialysis patients had detectable HIA. There were no differences in gender, ethnicity, disease states, or duration of hemodialysis. None of the patients had thrombocytopenia, and the incidence of thromboembolic events was statistically not different from patients without HIA. The non-CKD thrombocytopenic group had a lower incidence, 16/88 (18%) of positive HIA. CONCLUSIONS: Heparin-induced antibodies are relatively common in patients receiving hemodialysis. However, their presence does not appear to be associated with clinical consequences. Therefore, testing for HIA should be done only if a clinical indication is present. PMID- 19281737 TI - Absence of adynamic bone disease in African-Americans with CKD stage 5 after 3 years of vitamin D therapy guided by iPTH and the PTH-(1-84)/N-terminally truncated PTH fragments ratio. AB - BACKGROUND: Secondary hyperparathyroidism is a frequent complication of chronic kidney disease (CKD). The goal of treatment is to achieve circulating levels of parathyroid hormone (PTH) associated without oversuppression of bone turnover. This is commonly achieved by treatment with vitamin D analogs. Doses of vitamin D compounds are usually monitored by measurement of circulating levels of PTH. STUDY DESIGN: To prospectively assess the effects on bone histology of two different protocols for dosing vitamin D. SETTING AND PARTICIPANTS: African American patients from the same geographic area, managed by the same team of physicians in three dialysis clinics were studied. Patients were treated with vitamin D for 3 years and underwent bone biopsies for assessment of bone turnover. Dosing of vitamin D during the 3 years prior to the biopsy was done following two different guidelines. One group was treated following K/DOQI guidelines adapted to the bio-intact PTH assay (Protocol A), the other group was managed (Protocol B) following K/DOQI guidelines for intact PTH and/or the ratio of PTH-(1-84)/N-terminally truncated fragments (PTH ratio). PREDICTOR: Levels of circulating PTH and/or PTH ratio. OUTCOME: Prevalence of low bone turnover. MEASUREMENTS: Qualitative and quantitative assessment of bone histology after tetracycline labeling. RESULTS: 7 out of 22 patients managed following Protocol A were found to have low bone turnover (32%) by bone histology. None of the 21 patients managed by Protocol B for guidance of vitamin D therapy, had low bone turnover. LIMITATIONS: Lack of bone biopsy at the beginning of study. CONCLUSIONS: This report indicates that the additional information provided by the PTH ratio represents a distinct advantage in avoiding low bone turnover over the use of a single PTH assay to guide vitamin D dosing in African-American patients with CKD Stage 5 on dialysis. PMID- 19281738 TI - Comparison of intermittent intravenous and oral calcitriol in the treatment of secondary hyperparathyroidism in chronic hemodialysis patients: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Controversy exists regarding the optimal route and mode of calcitriol administration in the treatment of secondary hyperparathyroidism (SHPT). The aim of this study is to compare the efficacy and adverse reaction of calcitriol in the same mode (intermittent) but different route (intravenous versus oral) in the treatment of SHPT. METHODS: Electronic databases including Medline, Embase, Cochrane-controlled trials register, China Biological Medicine Disk (updated February 2008), and manual bibliographical searches were conducted. A meta analysis of all randomized controlled trials (RCTs) comparing intermittent intravenous with intermittent oral calcitriol in the treatment of SHPT was performed. RESULTS: 6 studies from 5 countries were finally considered for the meta-analysis, including a multicenter trial. The results of meta-analyses showed that there were no significant differences between the two routes in suppressing iPTH, AKP and rising Ca, P. In sensitivity analyses (baseline iPTH > or = 600 pg/ml, baseline iPTH < 600 pg/ml, and excluding the multicenter trial), the results were similar to those of the overall analyses. Results of the descriptive analysis showed that there were no significant differences between the two routes in the incidence of hypercalcemia and hyperphosphatemia. CONCLUSIONS: Our analysis indicated that there were no significant differences between intermittent intravenous and oral calcitriol in the treatment of SHPT, both in their efficacy and adverse reaction. Since the present studies had several limitations in methodological design and sample size, largescale and long-term comparisons of intermittent intravenous and oral treatment need to be done in a randomized, double-blind, placebo-controlled design to accurately evaluate their relative efficacy and safety. PMID- 19281740 TI - Use of two calcium concentrations in hemodialysis--report of a 20-year clinical experience. AB - Over the past almost 50 years several calcium concentrations in the dialysate (CaD) have been used to balance calcium in hemodialysis (HD) patients but a consensus as to which is most appropriate has not been established. Moreover, since the late 1980s, further confusion has been caused following the use of calcium salts as intestinal phosphate binders. This paper reports results of 387 chronic HD patients with respect to secondary hyperparathyroidism (sHPT) and renal osteodystrophy (ROD) of a single center over 20 years. The most important therapeutic measures applied were use of only 2 CaD, 1.5 and 1.75 mmol/l, with very few exceptions, administration of either calcium-containing or calcium magnesium-containing and/or calcium-free phosphate binders, no dietary restrictions and continuous compensation of uremic acidosis via dialysate and oral supplements of bicarbonate. Using one of the two CaD and selective administration of different phosphate binders for fine adjustment of serum calcium through this combination, we were able to maintain in the long term almost physiological conditions. With exception of the phosphate metabolism, most physiological functions with regard to sHPT and ROD returned close to normal. As a result, the incidence of hypercalcemia, hypocalcemia, extraosseous, extravascular calcification, bone pain and spontaneous bone fractures was extremely low. We conclude that the clinical advantages of the therapeutic measures, above all precise balance of calcium homeostasis, in our investigation were demonstrated by high survival rates (92% after the first year on HD, 82% after 2, and 55% after 5 years), low incidence of cardiovascular fatalities (about 25%), and very low incidence of sHPT (mostly normal parathyroid hormone levels, 1 parathyrdoidectomy within 20 years). PMID- 19281741 TI - Churg-Strauss syndrome in a child with IgA nephropathy. AB - Churg-Strauss syndrome (CSS) is a rare systemic vasculitis associated with asthma and eosinophilia. Withdrawal of corticosteroids in asthmatic patients is known to be a powerful trigger for the development of CSS. Renal involvement in patients with CSS is commonly manifested as antineutrophil cytoplasm antibody-associated necrotizing crescentic glomerulonephritis, however, concomitant CSS and the nephrotic syndrome or IgA nephropathy are rare. We report a 12-year-old boy with CSS associated with IgA nephropathy that developed while tapering oral steroids. The patient had a history of the nephrotic syndrome and asthma. PMID- 19281739 TI - Lanthanum carbonate treatment, for up to 6 years, is not associated with adverse effects on the liver in patients with chronic kidney disease Stage 5 receiving hemodialysis. AB - AIMS: The efficacy and tolerability of the phosphate binder, lanthanum carbonate, have been evaluated in long-term comparative studies and subsequent open-label extensions. Animal studies show that lanthanum has a very low bioavailability and absorbed lanthanum is primarily excreted in bile. A specified subset of data from four Phase III clinical trials and subsequent extension studies is presented, in order to assess the effects of lanthanum carbonate on the liver. METHODS AND MATERIALS: Hepatic biochemical tests for alanine transaminase, aspartate aminotransferase, alkaline phosphatase and bilirubin were performed. Adverse events classified as "liver and biliary system events" were recorded. RESULTS: In the four initial clinical trials, lanthanum carbonate was not associated with any adverse changes in transaminases or bilirubin. The incidence and nature of adverse events associated with the liver during lanthanum carbonate treatment was similar to that in the comparator groups. For patients who enrolled into the subsequent long-term follow-up study (up to 6 years of treatment), changes in transaminases were not clinically relevant and mean values were similar to those observed in the earlier trials. Overall, there was no increase in the incidence of adverse events associated with the liver reported after up to 6 years of treatment when compared with the results of the initial studies. CONCLUSIONS: There was no evidence of adverse effects of lanthanum carbonate on the liver in patients who received treatment for up to 6 years. PMID- 19281742 TI - Crescentic glomerulonephritis associated with miliary tuberculosis. AB - We report a 70-year-old man who presented with cervical lymphadenopathy, hypercalcemia, and acute renal failure. Lymph node biopsy revealed caseating granulomas. Renal biopsy showed immune complex-mediated diffuse proliferative glomerulonephritis with crescentic formation. Ultrastructural features of the glomerular disease were compatible with the description in infectious glomerulonephritis. Disseminated tuberculosis was diagnosed when sputum, urine, and bone marrow cultures confirmed the presence of Mycobacterium tuberculosis. Successful treatment with antituberculous agents accompanied by recovery of renal function. As far as we know, the association of tuberculosis with glomerulonephritis appears to be exceedingly rare. In addition to describing this case, we also review cases of tuberculosis-associated glomerulonephritis retrieved from the literature. PMID- 19281743 TI - Progressive renal failure as the first manifestation of monoclonal light-chain deposition disease with rapid multiple organ involvement. AB - Light-chain deposition disease (LCDD) is an uncommon condition in which monoclonal light chains are deposited in different organs causing varying degrees of tissue damage. We report the case of a 56-year-old male with progressive renal failure as the first manifestation of LCDD without initial evidence of monoclonal immunoglobulin protein in either serum or urine. The patient later developed severe systemic disease with rapid multiple organ involvement. Finally, he died 2 months after diagnosis, despite steroid and cyclophosphamide therapy. PMID- 19281744 TI - Pneumococcal endoprosthetic pulmonary valve endocarditis with crescentic glomerulonephritis and acute renal failure. AB - We describe a case of acute renal failure with crescentic glomerulonephritis, due to pneumococcal infective endocarditis on an endoprosthetic pulmonary valve. The patient's renal insufficiency subsequently improved following eradication of the microbe with antibiotics alone. Moreover, this is the first description of pneumococcal PVE leading to a crescentic glomerulonephritis. PMID- 19281745 TI - Atypical Alport syndrome associated with a novel COL4A5 mutation. AB - Alport syndrome is a progressive hereditary renal disease. Mutations in the genes encoding for three members of the type IV collagen protein family have been found to be the cause of the disease. Alport syndrome is often associated with sensorineural hearing loss and ocular abnormalities, and patients suffering from typical Alport syndrome usually develop end stage renal disease during adolescence or young adulthood. Here we report on a family with atypical Alport disease initially presenting as hereditary focal and segmental glomerulosclerosis. Genetic testing identified a previously undescribed COL4A5 mutation as cause of the disease. PMID- 19281746 TI - Aliskiren-associated acute renal failure with hyperkalemia. AB - We report the first case of acute renal failure with hyperkalemia associated with the recently marketed direct renin inhibitor aliskiren. To optimize blood pressure control, the antihypertensive medication of a 76-year-old hypertensive female patient was changed from the angiotensin II receptor antagonist irbesartan to aliskiren. Spironolactone was continued, as serum creatinine and potassium levels were initially normal. Two weeks later the patient presented with acute oliguric renal failure, symptomatic hyperkalemia and metabolic acidosis, necessitating emergency dialytic treatment. Unrecognized pre-existing renal insufficiency (CKD Stage 2 - 3) and the continuation of spironolactone were identified as predisposing risk factors. PMID- 19281747 TI - A case of acute renal failure due to hydroxyethyl starch. AB - A 73-year-old nephrotic female developed acute renal failure (ARF) with serum creatinine to 586 umol/l after 4 days of therapy with hydroxyethyl starch (HES). Renal biopsy demonstrated that the histopathological appearance was mesangioproliferative glomerulonephritis with tubulointerstitial changes resembling acute tubulointerstitial nephritis. "Pulse" therapy with methylprednisolone, hemodialyses and other symptomatic treatment were performed in the patient during the oliguric phase of the disease. There was no worsening of her renal function, and subsequently it recovered fully. After 6 months of follow-up she had a serum creatinine of 112 umol/l, proteinuria had declined to 0.18 g/day and she is now doing well. Our findings suggest that an immune disease due to a hapten induced by HES may be a possible factor in the pathogenesis of ARF. Therefore, it is important for clinicians to bear in mind the possibility of ARF due to HES and it has been proposed that, whenever possible, HES should not be used in idiopathic nephritic syndrome patients to avoid severe complications. PMID- 19281748 TI - Sequential occurrence of life-threatening hypokalemia and rebound hyperkalemia associated with barbiturate coma therapy. AB - OBJECTIVE: To report a sequential occurrence of life-threatening hypokalemia and rebound hyperkalemia following barbiturate coma therapy. CASE HISTORY: A 53-year old man was admitted to the division of nephrology due to sudden development of severe hypokalemia. The patient had been treated following a clinical diagnosis of traumatic subarachnoid hemorrhage and subdural hematoma. Barbiturate coma therapy had been performed on this patient. He developed fatal hypokalemia 10 hours after the start of thiopental administration which did not respond to potassium supplementation. The lowest potassium level following barbiturate coma therapy was 1.0 mmol/l. Severe bradycardia and cardiac arrest developed, which necessitated cardiac massage and treatment with epinephrine and atropine. He recovered from cardiac arrest. When thiopental infusion was suddenly stopped, the potassium level increased to 8.9 mmol/l, which required quick administration of calcium gluconate and infusion of glucose solution mixed with regular insulin. Despite such management, he developed asystole. After direct current cardioversion and emergency hemodialysis, he recovered from cardiac arrest and his serum potassium level was stabilized. CONCLUSION: We recommend that clinicians must be aware of the potential occurrence of severe hypokalemia, which is rare but fatal, following barbiturate coma therapy. Rebound hyperkalemia, which is fatal, may also occur following cessation of thiopental infusion. Clinicians should also be aware of this potential complication. Further studies are needed to elucidate the precise mechanism of this clinical event. PMID- 19281749 TI - "Nearly a stairway to heaven"--severe dichromate intoxication in a young man. AB - A 22-year-old man presented with dichromate intoxication in a suicidal attempt. He exhibited signs of liver and renal toxicity and very high serum chromium levels. Since it has been reported in the literature that hemodialysis and hemoperfusion are not sufficient to remove chromium, we tried plasmapheresis considering the fact that chromium salts bind to protein. Five plasmapheresis treatment sessions significantly lowered his serum and urinary chromium concentrations. The patient survived without organ damage despite ingestion of a lethal dichromate dose and high serum chromium concentration. Thus, plasmapheresis should be considered as a potential therapeutic option to reduce chromium concentrations. PMID- 19281750 TI - Wunderlich syndrome during antiplatelet drug therapy. AB - AIMS: Spontaneous hemorrhage in the kidney, also known as Wunderlich Syndrome, is a rare clinical problem. The most common cause of spontaneous renal hemorrhage is tumor. Other causes are rupture of a renal cyst, vasculitis, hydronephrosis, preeclampsia, and kidney infections. CASE HISTORY: A 67-year-old man was admitted complaining left flank colic pain. A contrast CT scan showed the presence of a subcapsular hematoma of the left kidney extending from the upper to the lower pole. He had no history of trauma and immunological screening tests for vasculitis were normal. His current therapy included acetylsalicylic acid (100 mg/daily) and lisinopril (20 mg/ daily). The patient was hospitalized for 4 days and his circulatory state remained stable. Nine months later an ultrasound examination showed complete resolution of the hematoma. One year later the patient was admitted again because of a spontaneous right calf hematoma and a thoroughly investigation of his coagulation pattern was carried out. Laboratory finding revealed a platelet defect, as the number of adenine nucleotides and other marker related to platelet activation were increased: ADP 1 mcM lack 2 masculine wave, ADP 2 mcM lack 2 masculine wave, Adrenalina 5 mcM lack 2 masculine wave, Adrenalina 10 mcM lack 2 masculine wave. Platelet activation markers: Gp53 in lysosomal membrane 0.48 Dpar (0 - 0.26). CONCLUSION: Our case describes the recurrence of spontaneous hemorrhages (perirenal and intramuscular hematoma) as a result of an underlying platelet aggregation defect worsened by administration of acetylsalicylic acid. In patients on antiplatelet treatment with a history of excessive bleeding a thorough investigation of coagulation status appears beneficial. PMID- 19281751 TI - A patient with pregnancy-related acute abdomen after hemodialysis for over 18 years. AB - Because pregnancy is rare in women with end-stage renal disease, dialysis patients have not been reported to present with acute abdominal symptoms related to pregnancy including ectopic pregnancy. A 41-year-old woman treated with hemodialysis for over 18 years was brought to the emergency room at our institution because of acute abdominal pain. Ultrasonography detected an abdominal fluid collection, and her anemia had worsened (hematocrit 18%). Emergency laparoscopic exploration disclosed a hemorrhagic corpus luteum of pregnancy, causing ovarian bleeding on the left. Coagulation of bleeding points was carried out. At this time, pregnancy at 7 weeks of gestation was discovered. After the procedures, hemodialysis frequency was increased to 5 times weekly, and an erythropoietin derivative was administered to maintain a hematocrit above 30%. The patient developed no hypertension. At 33 weeks of gestation, cesarean section was performed because of a decrease in amniotic fluid and frequent late deceleration of the fetal heart rate. A live baby girl weighing 1,422 g was born. The successful pregnancy reflects remarkable progress in dialysis technology. Pregnancy, then, can underlie an acute abdomen in childbearing-age women (14 - 44 years old) undergoing long-term dialysis. PMID- 19281752 TI - Achromobacter xylosoxidans, an emerging pathogen in catheter-related infection in dialysis population causing prosthetic valve endocarditis: a case report and review of literature. AB - INTRODUCTION: Dialysis catheter-related infection is a major cause of morbidity and mortality in patients on dialysis. In recent years, there have been reported cases of infections with opportunistic environmental organism, Achromobacter xylosoxidans (AX) causing bacteremia in patients on dialysis. However, no previous such reports on prosthetic valve endocarditis in a dialysis patient with Achromobacter xylosoxidans were found after a Medline search. We report such a case and review the literature. CASE: A 69-year-old diabetic man with bioprosthetic aortic valve replacement developed end-stage renal disease following infective endocarditis with Staphylococcus epidermidis. Even though he was treated successfully for his endocarditis, he developed further bacteremia with AX from his peripherally inserted central catheter (PICC) and the line was removed. He had further episodes of bacteremia with AX while having dialysis with tunneled line and the line was also removed. He was re-admitted with pyrexia and vegetations both in mitral and prosthetic aortic valve confirmed with transesophageal echo. His antimicrobial therapy with etrapenum, tigecycline and cotrimoxazole failed. He had both mitral and prosthetic aortic valve replacements but postoperatively developed multiorgan failure and died despite the intensive support. DISCUSSION: Achromobacter xylosoxidans is an aerobic, Gram-negative bacillus and considered to be an opportunistic pathogen with low virulence. Infective endocarditis is a potentially lethal complication of bacteremia. The choice of appropriate antibiotic is crucial in these cases. AX strains are highly resistant to antibiotics. The organism is usually susceptible to antipseudomonal penicillins, carbapenems and trimethoprim-sulfamethoxazole. CONCLUSION: AX is an emerging pathogen in catheter-related infection in the dialysis population and, therefore, needs vigilance and prompt treatment. Antimicrobial treatment should include susceptibility and synergy testing. Removal of central intravenous catheter should also be considered at the time of early presentation in patients at high risks of developing infective endocarditis. PMID- 19281753 TI - Successful treatment with voriconazole of prolonged Paecilomyces lilacinus fungemia in a chronic hemodialyzed patient. AB - Paecilomyces lilacinus is a saprophytic mould which rarely causes infection in humans. We report a case of Paecilomyces lilacinus catheter-related fungemia in a chronic hemodialyzed patient. Blood cultures remained positive for 8 weeks. The infection was cured after eventual acceptance by the patient of oral voriconazole treatment for 6 weeks and removal of the tunneled catheter. The literature on Paecilomyces fungemia in humans is reviewed. PMID- 19281754 TI - Management of abdominal aortic and iliac artery aneurysms by stent-graft implantation in a patient on CAPD. AB - We present the case of a 76-year-old male patient, who - after 2.5 years of CAPD treatment - underwent aorto-biiliac aneurysm reconstruction for aorto-biiliac aneurysm by bifurcational stent-graft implantation. To our knowledge this is the first case presentation of a stent-graft implantation and uninterrupted continuation of CAPD treatment in a patient on peritoneal dialysis. PMID- 19281755 TI - Influence of a supplementation with vitamins on cardiovascular morbidity and mortality in patients with end-stage renal disease: design and baseline data of a randomized clinical trial. PMID- 19281756 TI - Erythropoietin, heme oxygenase-1 and vascular tissue protective effects. PMID- 19281759 TI - Active DNA demethylation and DNA repair. AB - DNA methylation on cytosine is an epigenetic modification and is essential for gene regulation and genome stability in vertebrates. Traditionally DNA methylation was considered as the most stable of all heritable epigenetic marks. However, it has become clear that DNA methylation is reversible by enzymatic "active" DNA demethylation, with examples in plant cells, animal development and immune cells. It emerges that "pruning" of methylated cytosines by active DNA demethylation is an important determinant for the DNA methylation signature of a cell. Work in plants and animals shows that demethylation occurs by base excision and nucleotide excision repair. Far from merely protecting genomic integrity from environmental insult, DNA repair is therefore at the heart of an epigenetic activation process. PMID- 19281760 TI - The contribution of different cell lineages to bone repair: exploring a role for muscle stem cells. AB - An anabolic response driven by osteoblasts is critical for the process of bone healing. Current evidence suggests that these osteoblasts may arise from multiple tissue types and cell lineages. Stem cells present in the bone marrow, periosteum, local soft tissues, vasculature, and/or circulation have been shown to have osteogenic potential. Transplanted cells from these sources have also been shown to incorporate into induced ectopic bone or repaired bone. While these experiments demonstrate the latent capacity of different lineages to assume an osteoblastic phenotype under pro-osteogenic conditions, the actual contribution of the different lineages to various repair situations in vivo remains unclear. This review explores the data arising from different bone formation and repair models. We propose a model suggesting that cells arising from the local tissues, particularly muscle cells, may play an important role in fracture repair under situations where the periosteal and/or bone marrow progenitor populations are depleted. PMID- 19281761 TI - NRSF downregulation induces neuronal differentiation in mouse embryonic stem cells. AB - Differentiation of embryonic stem (ES) cells into neurons is accompanied by global changes in transcriptional programs. One transcription factor that has been shown to be involved in neuronal differentiation is neuron restrictive silencing factor/RE1-silencing transcription factor (NRSF/REST). NRSF is a transcriptional repressor that silences the transcription of a large number of neuronal genes by binding to a 21-bp consensus DNA sequence, the RE1 binding site/neuron-restrictive silencer elements (RE1/NRSE), present in the regulatory regions of neuronal genes. The goal of the current study was to examine the role of NRSF during differentiation of ES cells into neurons. To do this, ShRNA construct was used to downregulate NRSF in undifferentiated ES cells. Our results show that although control ES cells required induction by retinoic acid (RA) to differentiate efficiently into neurons, downregulation of NRSF was sufficient to drive the ES cells down the neuronal lineage even in the absence of RA. This downregulation also led to increased expression of mature neuronal markers, and concomitantly decreased glial fibrillary acidic protein (GFAP) expression. The results suggest that NRSF downregulation increases the population of mature neurons at the expense of GFAP-positive cells. PMID- 19281762 TI - Differentiation and regenerative capacities of human odontoma-derived mesenchymal cells. AB - Regenerating human tooth ex vivo and biological repair of dental caries are hampered by non-viable odontogenic stem cells that can regenerate different tooth components. Odontoma is a developmental dental anomaly that may contain putative post-natal stem cells with the ability to differentiate and regenerate in vivo new dental structures that may include enamel, dentin, cementum and pulp tissues. We evaluated odontoma tissues from 14 patients and further isolated and characterized human odontoma-derived mesenchymal cells (HODCs) with neural stem cell and hard tissue regenerative properties from a group of complex odontoma tissues from 1 of 14 patients. Complex odontoma was more common (9 of 14) than compound type and females (9 of 14) were more affected than males in our set of patients. HODCs were highly proliferative like dental pulp stem cells (DPSCs) but demonstrated stronger neural immunophenotype than both DPSCs and mandible bone marrow stromal cells (BMSCs) by expressing higher levels of nestin, Sox 2 and betaIII-tubulin. When transplanted with hydroxyapatite/tricalcium phosphate into immunocompromised mice, HODCs differentiated and regenerated calcified hard tissues in vivo that were morphologically and quantitatively comparable to those generated by DPSCs and BMSCs. When transplanted with polycaprolactone (biodegradable carrier), HODCs differentiated to form new predentin on the surface of a dentin platform. Newly formed predentin contained numerous distinct dentinal tubules and an apparent dentin-pulp arrangement. HODCs represent unique odontogenic progenitors that readily commit to formation of dental hard tissues. PMID- 19281763 TI - AFAP120 regulates actin organization during neuronal differentiation. AB - During development, dynamic changes in the actin cytoskeleton determine both cell motility and morphological differentiation. In most mature tissues, cells are generally minimally motile and have morphologies specialized to their functions. In metastatic cancer, cells generally lose their specialized morphology and become motile. Therefore, proteins that regulate the transition between the motile and morphologically differentiated states can play important roles in determining cancer outcomes. AFAP120 is a neuronal-specific protein that binds Src kinase and protein kinase C (PKC) and cross-links actin filaments. Here we report that expression and tyrosine phosphorylation of AFAP120 are developmentally regulated in the cerebellum. In cerebellar cultures, PKC activation induces Src kinase-dependent phosphorylation of AFAP120, indicating that AFAP120 may be a downstream effector of Src. In neuroblastoma cells induced to differentiate by treatment with a PKC activator, tyrosine phosphorylation of AFAP120 appears to regulate the formation of the lamellar actin structures and subsequent neurite initiation. Together, these results indicate that AFAP120 plays a role in organizing dynamic actin structures during neuronal differentiation and suggest that AFAP120 may help regulate the transition from motile precursor to morphologically differentiated neurons. PMID- 19281764 TI - Endocytotic activity of bladder superficial urothelial cells is inversely related to their differentiation stage. AB - The composition of the apical plasma membrane of bladder superficial urothelial cells is dramatically modified during cell differentiation, which is accompanied by the change in the dynamics of endocytosis. We studied the expression of urothelial differentiation-related proteins uroplakins and consequently the apical plasma membrane molecular composition in relation to the membrane-bound and fluid-phase endocytosis in bladder superficial urothelial cells. By using primary urothelial cultures in the environment without mechanical stimuli, we studied the constitutive endocytosis. Four new findings emerge from our study. First, in highly differentiated superficial urothelial cells with strong uroplakin expression, the endocytosis of fluid-phase endocytotic markers was 43% lower and the endocytosis of membrane-bound markers was 86% lower compared to partially differentiated cells with weak uroplakin expression. Second, superficial urothelial cells have 5-15-times lower endocytotic activity than MDCK cells. Third, in superficial urothelial cells the membrane-bound markers are delivered to lysosomes, while fluid-phase markers are seen only in early endocytotic compartments, suggesting their kiss-and-run recycling. Finally, we provide the first evidence that in highly differentiated cells the uroplakin positive membrane regions are excluded from internalization, suggesting that uroplakins hinder endocytosis from the apical plasma membrane in superficial urothelial cells and thus maintain optimal permeability barrier function. PMID- 19281765 TI - Spheroids of granulosa cells provide an in vitro model for programmed cell death coupled to steroidogenesis. AB - We describe the use of rotary cultures (72 rpm) as an excellent method for generating spheroids from dispersed bovine granulosa cells (GC). The GC spheroids were symmetrical (diameter between 100 and 200 microm), easily accessible, and could be obtained at high yields. On day one, the spheroids showed a two-layered outer zone of cells that stained lighter than the inner zone in semi-thin sections. Bromodeoxyuridine (BrdU) uptake was frequent and randomly distributed. By day two, a striking decrease in BrdU uptake was noted. Apoptotic bodies appeared up to day four, as did TUNEL and propidium iodide labelled dead cells. At that time, the inner zone contained cells with large-sized vacuoles and the core was amorphous. The large-sized vacuoles were identified at the ultrastructural level and represented autophagosomes and autophagolysosomes that were in different stages of development. Surprisingly, conspicuous signs of cell death were accompanied by an increase in spontaneous luteinization compared to conventional stationary cultures. We detected high levels of progesterone (immunoassay) accompanied by high levels of the proteins and enzymes relevant for steroidogenesis (StAR, P450scc, 3beta-HSD by immunoblot and immunohistochemistry, respectively). CONCLUSIONS: Concomitant to cell death, GC spheroids augment progesterone synthesis. The GC spheroids provide an ideal model for studying steroidogenesis coupled to programmed cell death at the level of the mitochondria. PMID- 19281766 TI - Differentiation-dependent modification and subcellular distribution of aquaporin 0 suggests multiple functional roles in the rat lens. AB - Using immunohistochemistry and mass spectrometry, differentiation-dependent changes in the subcellular distribution and processing of aquaporin-0 (AQP0) have been mapped in the rat lens. Sections labelled with C-terminal tail AQP0 antibodies yielded two concentric rings of labelling with minimal signal in the lens core. The rings were separated by a transient zone of decreased labelling located prior to the transition of differentiating fiber (DF) cells into mature denucleated fiber (MF) cells. Mass spectrometry showed that the loss of core labelling was due to AQP0 cleavage, while the transient loss of labelling was more likely caused by masking of the antibody epitope. AQP0 subcellular distribution changed with radial distance into the lens. In peripheral DF cells, AQP0 was found throughout both broad and narrow side membranes. In deeper-lying DF cells, AQP0 aggregated into plaque-like structures located on the broad sides. This shift occurred prior to the transient loss of AQP0 signal, and coincided with formation of broad-side membrane invaginations between adjacent fiber cells to which filensin, a known binding partner of AQP0, was also localized. After nuclei loss, AQP0 was once again distributed throughout MF cell membranes. In the absence of protein synthesis, the observed subcellular redistribution of AQP0 in DF and subsequent cleavage of AQP0 in MF are suggestive of a switch in the function of AQP0 from a water channel to a junctional protein. PMID- 19281767 TI - Differences in goblet cell differentiation between Crohn's disease and ulcerative colitis. AB - Goblet cells are mucin-secreting intestinal cells forming the mucus layer that protects the mucosal surface. Ulcerative colitis (UC) has been associated with a defective colonic mucus layer and a reduced number of goblet cells. In experimental animals, colonic goblet cell differentiation is regulated by interacting transcription factors Hath1, KLF4 and the Notch, as well as Wnt pathways, whereas data in humans are limited. We investigated goblet cell differentiation factors and mucins in controls and in inflammatory bowel diseases (IBDs). We performed real-time PCR for Hath1, KLF4, several ligands, receptors and target genes of the Notch and Wnt pathways, as well as several mucins in biopsies from the sigmoid colon of controls (n=21), Crohn's disease (CD, n=48) and UC (n=40). In addition, Hath1 protein was quantitated with Western blot and localized with immunohistochemistry. Notably, the degree of inflammation as measured by IL-8 and histology was similar in both disease entities. The proportion of goblet cells was lowered in both IBDs, but specifically diminished in the upper third of the crypt in UC. Comparable levels of inflammation induced both Hath1 (2.0-fold, p<0.001) and KLF4 (1.8-fold for KLF4, p=0.031) mRNA expression in CD but not in UC (0.8-0.9-fold, ns). The differential induction was confirmed for Hath1 protein using Western blot. Hath1 immunostaining was found mostly in the lower half of the colonic crypts. Hath1, KLF4 and the Notch target gene Hes1 were significantly (p<0.001) and positively correlated. Moreover, both Hath1 and KLF4 were correlated (p<0.001) with MUC1, MUC2 as well as MUC4 in all control and IBD cohorts. The results indicate that both transcription factors are key regulators of goblet cell differentiation and mucin formation in the human colon. Conspicuously, inflammation is associated with an enhanced goblet cell differentiation in CD but not in UC, a defect possibly of pathogenic importance. PMID- 19281768 TI - Urothelial transdifferentiation to prostate epithelia is mediated by paracrine TGF-beta signaling. AB - The embryonic urogenital sinus mesenchyme (UGM) induces prostate epithelial morphogenesis in development. The molecular signals that drive UGM-mediated prostatic induction have not been defined. We hypothesized that the TGF-beta signaling directed the prostatic induction. UGM from TGF-beta type II receptor stromal conditional knockout mice (Tgfbr2(fspKO)) or control mice (Tgfbr2(floxE2/floxE2)) was recombined with wild-type adult mice bladder urothelial cells. The resulting urothelium associated with Tgfbr2(floxE2/floxE2) UGM was instructively differentiated into prostatic epithelium, as expected. In contrast, the urothelium associated with Tgfbr2(fspKO) UGM permissively maintained the phenotype of bladder epithelial cells. Microarray analysis of UGM tissues suggested the down-regulation of multiple Wnt ligands and the up regulation of the Wnt antagonist, Wif 1, by the Tgfbr2(fspKO) UGM compared with Tgfbr2(floxE2/floxE2) UGM. The overexpression of Wif-1 by wild-type UGM resulted in the inhibition of prostatic induction. These data suggest that the stromal TGF beta activity mediated by paracrine Wnt is necessary for the induction of prostatic differentiation. As Wnt ligands mediate differentiation and maintain the stem cell phenotype, the contribution of mouse stem cells and somatic cells to prostatic epithelium in the tissue recombination models was tested. The directed differentiation of mouse embryonic stem cells by UGM is suggested by a threshold number of mouse stem cells required in prostatic differentiation. To determine the contribution of somatic cells, the adult bladder epithelial compartment was labeled with green-fluorescent vital dye (CMFDA) and the stem like cells marked by bromodeoxyuridine (BrdU) label-retention. The resulting prostatic epithelia of the tissue recombinants maintained the CMFDA dye, suggesting minimal cell division. Thus, the UGM can induce endoderm-derived epithelia and stem cells to form prostate through a transdifferentiation mechanism that requires stromal TGF-beta signaling to mediate epithelial Wnt activity. PMID- 19281769 TI - Simultaneous haploinsufficiency of Pten and Trp53 tumor suppressor genes accelerates tumorigenesis in a mouse model of prostate cancer. AB - Tumor suppressor gene PTEN is important in the initiation and progression of human prostate carcinoma, whereas the role of TP53 remains controversial. Since Pten/Trp53 double conditional knockout mice show earlier onset and fast progression of prostate cancer when compared to Pten knockout mice, we asked whether heterozygosity of these two tumor suppressor genes was sufficient to accelerate prostatic tumorigenesis. To answer this question we examined prostatic lesion progression of Pten/Trp53 double heterozygous mice and a series of controls such as Pten heterozygous, Pten conditional knockout, Trp53 heterozygous and Trp53 knockout mice. Tissue recombination of adult prostatic epithelium coupled with embryonic rat seminal vesicle mesenchyme was used as a tool to stimulate prostatic epithelial proliferation. In our study, high-grade prostatic intraepithelial neoplasia (PIN) was found with high frequency at 8 weeks post tissue recombination transplantation. PIN lesions in Pten/Trp53 double heterozygous mice were more severe than those seen in Pten heterozygous alone. Furthermore, morphologic features attributable to Pten or Trp53 loss appeared to be enhanced in double heterozygous tissues. LOH analysis of Pten and Trp53 in genomic DNA collected from high-grade PIN lesions in Pten heterozygous and Pten/Trp53 double heterozygous mice showed an intact wild-type allele for both genes in all samples examined. In conclusion, simultaneous heterozygosity of Pten and Trp53 accelerates prostatic tumorigenesis in this mouse model of prostate cancer independently of loss of heterozygosity of either gene. PMID- 19281770 TI - New aspects of adipogenesis: radicals and oxidative stress. AB - Preadipocytes are multipotent adipogenic precursor cells that can be isolated from mature adipose tissue. They have been receiving increasing attention in the context of obesity, type 2 diabetes, and other nutrition-associated diseases. Understanding the physiological and pathophysiological processes in fat neo formation, energy homeostasis, and adipose tissue physiology is the basis for research on metabolic diseases and the respective pharmaceutical intervention. While the hormonal influence on intracellular signaling in adipogenesis has been intensively investigated, the effects of free radical formation and oxidative stress have just started to gain scientific attention. This review summarizes the present knowledge on the main molecular pathways in preadipocyte maturation and focuses on recent findings indicating that besides hormonal stimuli reactive oxygen species (ROS) and free radicals may also interact with preadipocyte differentiation. PMID- 19281771 TI - Emerging cues mediating astroglia lineage restriction of progenitor cells in the injured/diseased adult CNS. AB - Other than specific neurogenic regions, the adult central nervous system (CNS) is not conducive for neuronal regeneration and neurogenesis, particularly at sites of injury or neurodegeneration. Engraftment of neural stem/progenitor cells into non-neurogenic regions or sites of injury/disease invariably results mainly in astroglia differentiation. The reasons for such a lineage restriction have not been well defined. Recent findings have brought to light some underlying novel mechanistic basis for this preferential differentiation into astroglia. The more oxidized state of pathological brain tissue leads to upregulation of the protein deacetylase sirtuin 1 (Sirt1). Sirt1 appears to stabilize a co-repressor complex of Hairy/enhancer of split (Hes)1, thereby suppressing expression of the proneuronal transcription factor Mash1, and directs progenitor cell differentiation towards the glia lineage. Sirt1 upregulated by CNS inflammation may also inhibit neuronal differentiation. Myelin-associated inhibitors such as Nogo, acting through the Nogo-66 receptor (NgR), also appear to promote neural stem/progenitor cell differentiation into astrocytes. Understanding the molecular basis of glia lineage restriction of neural progenitors in the injured or diseased CNS would provide handles to improving the success of stem cell-based transplantation therapy. PMID- 19281772 TI - Generation of living color transgenic zebrafish to trace somatostatin-expressing cells and endocrine pancreas organization. AB - In the present study, both gfp and rfp transgenic zebrafish lines using a 2.5-kb zebrafish somatostatin2 (sst2) promoter were generated. During embryonic development, expression of GFP/RFP in the endocrine pancreas of transgenic embryos was initiated at approximately 20 hpf and the number of GFP/RFP positive cells in the pancreas increased in subsequent stages; thus, our newly generated Tg(sst2:gfp) and Tg(sst2:rfp) lines faithfully recapitulated sst2 expression in endocrine pancreatic cells and provided a useful tool in analyzing the development of Sst2-producing delta-cells in the pancreas. By crossing these new transgenic lines with previously available transgenic lines targeted in insulin (Ins)-producing beta-cells, Tg(ins:gfp) and Tg(ins:rfp), in combination with immunodetection of glucagon (Gcg)-producing alpha-cells and pancreatic polypeptide (PP)-producing PP-cells, the organization and composition of endocrine islets were investigated in both embryonic and adult pancreas. We found that there was always a big cluster of endocrine cells (principal islet) in the anterior-dorsal pancreas, followed by numerous smaller clusters (variable in size) of endocrine cells (secondary islets) along the anterior-posterior axis of the pancreas. All four types of endocrine cells were found in the principal islet, but secondary islets may or may not contain PP-cells. In addition, there were also discrete endocrine cells throughout the pancreas. In all co localization experiments, we did not find any endocrine cells positive for more than one hormone markers, suggesting that these endocrine cells produce only a single hormone. In both principal and secondary islets, we found that beta-cells were generally located in the center and non-beta cells in the periphery; reminiscent of the "mantel-core" organization of islets of Langerhans in mammals where beta-cells form the core and non-beta-cells the mantel. In zebrafish primary islet, beta-cells constitute most of the mass (approximately 50%), followed by delta-cells and alpha-cells (20-25% each), and PP-cells (1-2%); this is also similar to the composition of mammalian islets. PMID- 19281773 TI - Developmental pathways during in vitro progression of human islet neogenesis. AB - Islet neogenesis, or the differentiation of islet cells from precursor cells, is seen in vitro and in vivo both embryonically and after birth. However, little is known about the differentiation pathways during embryonic development for human pancreas. Our previously reported in vitro generation of islets from human pancreatic tissue provides a unique system to identify potential markers of neogenesis and to determine the molecular mechanisms underlying this process. To this end, we analyzed the gene expression profiles of three different stages during in vitro islet generation: the Initially Adherent, Expanded, and Differentiated stages. Samples from four human pancreases were hybridized to Affymetrix U95A GeneChips, and data analyzed using GeneSpring 7.0/9.0 software. Using scatter plots we selected genes with a 2-fold or greater differential expression. Of the 12,000 genes/ESTs present on these arrays, 295 genes including 38 acinar-enriched genes were selectively lost during the progression from the Initially Adherent stage to the Expanded stage; 468 genes were increased in this progression to Expanded tissue; and 529 genes had a two-fold greater expression in the Differentiated stage than in the Expanded tissue. Besides the expected increases in insulin, glucagon, and duct markers (mucin 6, aquaporin 1 and 5), the beta cell auto-antigen IA-2/phogrin was increased 5-fold in Differentiated. In addition, developmentally important pathways, including notch/jagged, Wnt/frizzled, TGFbeta superfamily (follistatin, BMPs, and SMADs), and retinoic acid (COUP-TFI, CRABP1, 2, and RAIG1) were differentially regulated during the expansion/differentiation. Two putative markers for islet precursor cells, UCHL1/PGP9.5 and DMBT1, were enhanced during the progression to differentiated cells, but only the latter could be a marker of islet precursor cells. We suggest that appropriate manipulation of these differentiation-associated pathways will enhance the efficiency of differentiation of insulin-producing beta-cells in this in vitro model. PMID- 19281774 TI - Role of NEDD8 in HIV-associated lipodystrophy. AB - The pathogenetic bases of HAART-associated lipodystrophy are still poorly known, even if it is clear that adipose tissue and its metabolism are sensitive to antiretroviral therapy alone and/or in combination with HIV infection. The NEDD8 system is essential for the regulation of protein degradation pathways involved in cell cycle progression, morphogenesis and tumorigenesis. We investigated the possible involvement of NEED8 in adipogenesis and, consequently, in HIV-related lipodystrophy. One hundred HIV-1-infected patients were included in the study. Using an in vitro model of adipogenesis we evaluated the effects on adipogenesis of the forced expression of NEDD8 together with efavirenz, stavudine, saquinavir, amprenavir and indinavir, belonging to the three main classes of anti-HIV medications. We showed that NEDD8 expression level is higher in the peripheral blood of HIV patients developing lipodystrophy. Coherently, forced expression of NEDD8 in an in vitro model of adipogenesis was able to perturb expression of some key proteins involved in adipogenesis, such as C/EBPalpha and PPARgamma, possibly acting throughout the NEDD8/p27/beta-catenin pathway. Moreover, three out of five evaluated drugs were able to affect adipocyte differentiation: efavirenz, stavudine and saquinavir. Finally, we have shown that NEDD8 was expressed in the fat tissue of lipodystrophic patients, being significantly higher in the lipodystrophic patients with respect to the controls, thus further confirming the altered NEDD8 expression in the fat tissue of HIV-infected patients affected by lipodystrophy. Taken together, our data support the hypothesis of an implication of NEDD8 through p27 and beta-catenin pathways in the disruption of adipogenesis and consequent lipodystrophy in patients affected by HIV infection under HAART therapy with qualitative and quantitative differences according to diverse antiretroviral treatments. These evidences indicate the NEDD8/beta-catenin/p27 pathway as a possible molecular target for prevention of lipodystrophy development in patients under HAART therapy. PMID- 19281775 TI - RhoA promotes differentiation of WB-F344 cells into the biliary lineage. AB - When cultured on Matrigel, liver precursor epithelium WB-F344 cells could be induced to differentiate into biliary cells in which RhoA expression was upregulated. To further investigate the role of RhoA in WB cell differentiation initiated by Matrigel treatment, we constructed constitutively active RhoA expressing vectors and stably transfected them into WB-F344 cells. Accompanying upregulation of biliary lineage markers and morphological changes, cells with ectopically active RhoA expression were found to form bile-duct-like structures even without Matrigel treatment. Besides, ROCK inhibitor Y27632 treatment eliminated luminal morphogenesis. F-actin cytoplasmic staining further verified that the RhoA-ROCK signal pathway was involved in differentiation of WB cells into the biliary lineage. In conclusion, our results suggested that the RhoA-ROCK stress fibre system plays an obligatory role in Matrigel-induced WB-F344 cell luminal morphogenesis and further differentiation. PMID- 19281776 TI - Expression and function of HOXA genes in normal and neoplastic ovarian epithelial cells. AB - We studied the roles of three HOXA genes in cultured normal ovarian surface epithelial (OSE) cells and ovarian cancer cells. They included HOXA4 and HOXA7 because, by cDNA microarray analysis, these were more highly expressed in invasive ovarian carcinomas than in benign or borderline (noninvasive) ovarian tumors, and HOXA9 because it characterizes normal oviductal epithelium, which resembles ovarian serous adenocarcinomas. The three HOXA genes were more highly expressed when OSE cells were dividing and motile than when they were confluent and stationary, and also when they dispersed in response to EGF treatment or to reduced calcium concentrations in culture media. The expression of the HOXA genes varied among ovarian cancer cell lines, but was highest in lines with compact epithelial morphologies. We focused on HOXA4 as the most highly expressed in the ovarian carcinoma array. HOXA4 expression did not parallel proliferative activities of either OSE or ovarian cancer lines. Moreover, modifying HOXA4 expression in ovarian cancer cell lines did not alter either E-cadherin expression or CA125 secretion. However, HOXA4 downregulation enhanced EGFR phosphorylation and migration in serum-starved OSE and ovarian cancer cells in response to EGF, and enhanced migration of all ovarian cancer lines in 5% serum even without EGF treatment. Thus, HOXA4 expression does not correlate with proliferation or with epithelial differentiation, but it increases in response to OSE cell dispersion and negatively regulates EGFR activation and the motility of OSE and of ovarian cancer cells. HOXA4 expression was highest in cancer lines with compact epithelial growth patterns, suggesting, again, an anti-dispersion function. In summary, increased HOXA4 expression in ovarian cancer appears to constitute a tumor-suppressive, homeostatic response to aberrant cell behavior, and, in particular, to cell dispersion and migration. PMID- 19281777 TI - Identification of an aberrant cell line among human adipose tissue-derived stem cell isolates. AB - Adipose tissue-derived stem cells (ADSC) are isolated from the stromal vascular fraction (SVF) of adipose tissue and considered an excellent cell source for regenerative medicine. During the isolation and propagation of several human ADSC cell lines, we observed the emergence of an unusual cell line designated HADSC-6. Although initially fibroblast-like as typical ADSC are, HADSC-6 cells became homogeneously cuboid in shape, had very little cytoplasm, and formed aggregates with capsule-like boundary. Proliferation assay showed that HADSC-6 grew much faster than typical HADSC cell lines, such as HADSC-20. Immunocytochemistry showed that HADSC-6 did not express endothelial markers CD31 and vWF, and matrigel tube formation assay showed that it was unable to form endothelial-like tube structures. However, LDL uptake, a reliable endothelial marker, was positively identified. Chromosomal analysis showed that HADSC-6 cells were hypertriploid, and soft agar colony formation assay showed that they were able to proliferate and form large colonies in an anchorage-independent manner. However, tumorigenicity test showed that HADSC-6 was unable to form tumors in athymic mice. RT-PCR analysis showed that both HADSC-6 and HADSC-20 expressed VEGF-A, VEGF-B, VEGF-D, and VEGFR1 but not VEGFR2 or VEGFR3. VEGF-C, however, was expressed at a high level in HADSC-20 but undetectable in HADSC-6. In the IGF system, IGF-1 was abundantly expressed in HADSC-20 but marginally detectable in HADSC-6, and IGF-1R was abundantly expressed in HADSC-6 but not detectable in HADSC-20. In the FGF system, bFGF was abundantly expressed in HADSC-20 but marginally detectable in HADSC-6, and FGFR1 was abundantly expressed in both. Taken together, these results suggested that HADSC-6 cells were spontaneously transformed from the endothelium; therefore, they were further compared to previously published data of four naturally occurring human angiosarcoma cell lines. The results showed that the established angiosarcoma cell lines exhibit considerable variations among themselves and HADSC-6 displayed most of these variable characteristics. PMID- 19281778 TI - A microRNA, miR-101a, controls mammary gland development by regulating cyclooxygenase-2 expression. AB - Mammary glands exhibit a series of developmental states that are typified by proliferation, differentiation, and involution. Here, we demonstrate that a microRNA (miRNA), miR-101a, plays an important role in the process of mammary gland development. We used miRNA microarray analysis to show that some miRNAs exhibit changes in their expression during mouse mammary gland epithelial cell (HC11) differentiation, which corresponds to the time when these cells acquire the milk-producing phenotype. In particular, we observed an increase of miR-101a expression throughout differentiation and involution in mammary gland tissue, as well as in HC11 cells. Overexpression experiments revealed that miR-101a suppressed the expression of beta-casein mRNA, a milk protein, and marker of cell differentiation, but its suppression was not mediated by transcriptional or direct post-transcriptional regulation of beta-casein mRNA. Overexpression of miR 101a also inhibited HC11 cell proliferation that could influence the differentiation state of the mammary gland. We speculate that a direct target of miR-101a is cyclooxygenase-2 (Cox-2) mRNA because there was an inverse relationship between these two genes during mammary gland development. Indeed, Cox-2 protein expression was suppressed by the overexpression of miR-101a, and the luciferase activity of reporter constructs containing the Cox-2 3'UTR was also suppressed by miR-101a overexpression. As Cox-2 has been shown to mediate cell proliferation, it is possible that the inhibition of HC11 cell proliferation by miR-101a might be mediated by Cox-2. Taken together, these results suggest that miR-101a regulates cell proliferation via altering Cox-2 expression, which is critical for controlling mammary gland development. PMID- 19281779 TI - Neurotransmitter responsiveness during early maturation of neural progenitor cells. AB - Neurotransmitters are potential regulators of proliferation and differentiation of neural progenitor cells (NPC). To gain insight into the dynamics of neurotransmitter responsiveness, neurospheres were prepared from the lateral ventricles of postnatal day 6/7 mice. Individual NPCs migrating out from spheres were simultaneously monitored using Ca(2+) imaging, during the initial 8 days of differentiation, at an area between the inner edge of the sphere and outer periphery of the area of migration. At the first day of differentiation most cells showed metabotropic responses (Ca(2+) discharge from stores) to glutamate (pharmacologically identified as metabotropic glutamate receptor 5, mGluR 5), norepinephrine (NE), acetylcholine (Ach) and ATP, and a smaller proportion of cells also responded to substance P (SP). When outside the neurosphere, many of mGluR5 responding cells gained immunostaining for markers of neuronal lineage (Tuj-1 and NeuN). The number of cells responding through mGluR5 (and responses to Ach, NE and SP) showed during subsequent days of differentiation (day 2-3 onwards) a decline with time and progressively disappeared at the outer periphery of the area of migration. Conversely the number ionotropic glutamate responses as well as responses to depolarization increased in this area. After 5-8 days of differentiation mGluR5 responses could only be observed at the very inner edge of the neurosphere. At 8 days the migrated cells showed very robust ionotropic responses to glutamate, NMDA and depolarization comparable to mature neurons. Taken together, the data presented here suggest that differentiation of NPCs is a dynamic process triggered by cell migration, which leads to a loss of regulatory influences imposed by the inner milieu of the neurosphere. The subsequent switch or loss of metabotropic responses to glutamate, SP, NE, Ach and ATP with the gain of excitable characteristics such as ionotropic responses appears to be a key event in the final differentiation process. PMID- 19281780 TI - Lumbo-sacral neural crest derivatives fate mapped with the aid of Wnt-1 promoter integrate but are not essential to kidney development. AB - Neural crest (NC) cells may be involved in kidney organogenesis by providing inductive signals and contributing to cells of the renal stroma. We show here that the lumbo-sacral NC cells fate mapped with the aid of Wnt-1 promoter in the mouse migrate close to the metanephros at the initiation of organogenesis but these cells remain superficial to the condensed Pax2-expressing mesenchymal cells. NC-derived cells enter later into the kidney proper from the midline region. The NC cells contribute also to development of the extra-adrenal para aortic bodies, Zuckerkandl's bodies and the nerve cord of the sympathetic nervous system. Splotch (Sp(2H)/Sp(2H)) embryos, having a NC defect in the lumbo-sacral region, develop a normal metanephros even though the kidney does not express the NC markers Sox10, Phox2b and tyrosine hydroxylase. Consistent with the histological findings, the kidneys of Sp(2H)/Sp(2H) embryos also express the stromal genes Foxd1, Hoxa10 and RARbeta normally. Wnt-1 promoter-marked wild-type LacZ NC cells migrate intensely from the heterologous inducer tissue of the embryonic dorsal spinal cord (SPC) to the kidney mesenchyme, but tubule induction does not depend on NC migration, since the Sp(2H)/Sp(2H) SPC also induces tubulogenesis. The Sp(2H)/Sp(2H) mesenchyme also remains competent for tubulogenesis. We conclude that the NC cells fate mapped with the aid of Wnt-1 promoter migrate to the close to the metanephros and form later derivatives integrating with the kidney, but they may not be essential to the development of the stromal cells nor they may provide critical morphogenetic signals to regulate early kidney development in vivo. PMID- 19281781 TI - Interactions between TGF-beta1 and TGF-beta3 and their role in medial edge epithelium cell death and palatal fusion in vitro. AB - In recent decades, studies have shown that both TGF-beta(1) and TGF-beta(3) play an important role in the induction of medial edge epithelium (MEE) cell death and palatal fusion. Many of these experiments involved the addition or blockage of one of these growth factors in wild-type (WT) mouse palate cultures, where both TGF-beta(1) and TGF-beta(3) are present. Few studies have addressed the existence of interactions between TGF-beta(1) and TGF-beta(3), which could modify their individual roles in MEE cell death during palatal fusion. We carried out several experiments to test this possibility, and to investigate how this could influence TGF-beta(1) and TGF-beta(3) actions on MEE cell death and palatal shelf fusion. We double-immunolabelled developing mouse palates with anti-TGF-beta(1) or anti TGF-beta(3) antibodies and TUNEL, added rhTGF-beta(1) or rhTGF-beta(3) or blocked the TGF-beta(1) and TGF-beta(3) action at different concentrations to WT or Tgf beta(3) null mutant palate cultures, performed in situ hybridizations with Tgf beta(1) or Tgf-beta(3) riboprobes, and measured the presence of TUNEL-positive midline epithelial seam (MES) cells and MES disappearance (palatal shelf fusion) in the different in vitro conditions. By combining all these experiments, we demonstrate great interaction between TGF-beta(1) and TGF-beta(3) in the developing palate and confirm that TGF-beta(3) has a more active role in MES cell death than TGF-beta(1), although both are major inductors of MES disappearance. Finally, the co-localization of TGF-beta(1), but not TGF-beta(3), with TUNEL in the MES allows us to suggest a possible role for TGF-beta(1) in MES apoptotic clearance. PMID- 19281782 TI - An epigenetic mechanism regulates germ cell-specific expression of the porcine Deleted in Azoospermia-Like (DAZL) gene. AB - The Deleted in Azoospermia-Like (DAZL) gene is specifically expressed in fetal and adult gonads. While DAZL is known to play a role during gametogenesis, the mechanisms governing its germ cell-specific expression remain unclear. We identified the 5' untranslated region (UTR) of the porcine DAZL gene and cloned and characterized 2 kilobase pairs of its TATA-less 5' flanking region, identifying CpG-rich regions within the proximal promoter. Nine of 18 CpG sites in proximity to one region were largely unmethylated in germ cells but hypermethylated in somatic cells, suggesting that DNA methylation may regulate DAZL promoter activity. Furthermore, DAZL expression was induced in fibroblasts treated with a demethylating agent. Deletion analyses revealed that the minimal 149 base pair promoter region was sufficient to activate transcription. In vitro methylation of a reporter construct corresponding to these 149 base pairs resulted in complete suppression of DAZL promoter activity in primordial germ cells, further supporting a role for methylation in regulating DAZL expression. Interestingly, the differentially methylated region was shown to harbor several putative Sp1-binding sites. Mutation of only the most highly conserved site significantly reduced promoter activity in a reporter assay. Furthermore, gel shift assays revealed that Sp1 was able to specifically bind to this site, and that complex formation was inhibited when CpG dinucleotides within this region were methylated. Chromatin immunoprecipitation (ChIP) assays revealed that in vivo Sp1 binding to the core DAZL promoter region was enriched in germ cells but not in fibroblasts. Our data suggests that DNA methylation may suppress DAZL expression in somatic cells by interfering with Sp1 binding. This study provides insights into the potential mechanisms underlying the regulation of germ cell specific gene expression. PMID- 19281783 TI - Degenerative muscle fiber accelerates adipogenesis of intramuscular cells via RhoA signaling pathway. AB - In some pathological conditions such as Duchenne muscular dystrophy, it has been known that a fatty infiltration in skeletal muscle is often observed and that is also one of primary factors to induce marked decline of muscular strength. However, the mechanism of fatty infiltration, cellular origin of accumulated adipocytes and its significance are not fully understood. The fact that persistent degenerative muscle fibers are present on dystrophic muscle leads us to hypothesize that muscle fiber condition affects fatty infiltration in skeletal muscle. We employed a single fiber culture system to determine whether fiber condition affects an appearance of adipocytes on the fibers. Artificially hyper contracted muscle fibers (HCF), generated from isolated intact fibers (IF) of rat extensor digitrum longus muscle, were maintained as non-adherent cultures for 5-7 days. Interestingly, there appeared to be considerable numbers of mature adipocytes on HCF, whereas no adipocytes were seen on IF, indicating that cells on HCF spontaneously differentiated into mature adipocytes. Activation of RhoA signaling by the addition of thrombin decreased the number of adipocytes on HCF in a dose-dependent manner, whereas the number of MyoD-positive myoblasts increased. In contrast, Y-27632, a specific inhibitor of Rho kinases (ROCK), induced adipogenic differentiation of cells derived from IF. In addition, administration of Y-27632 into mouse regenerating muscle resulted in fat accumulation in the muscle. Taken together, the present studies clearly demonstrated that muscle fiber condition affects fat accumulation in skeletal muscle and that is possibly mediated by the RhoA signaling pathway. PMID- 19281784 TI - Alpha actin isoforms expression in human and rat adult cardiac conduction system. AB - In the adult heart, cardiac muscle comprises the working myocardium and the conduction system (CS). The latter includes the sinoatrial node (SAN), the internodal tract or bundle (IB), the atrioventricular node (AVN), the atrioventricular bundle (AVB), the bundle branches (BB) and the peripheral Purkinje fibers (PF). Most of the information concerning the phenotypic features of CS tissue derives from the characterization of avian and rodent developing hearts; data concerning the expression of actin isoforms in adult CS cardiomyocytes are scarce. Using specific antibodies, we investigated the distribution of alpha-skeletal (alpha-SKA), alpha-cardiac (alpha-CA), alpha smooth muscle (alpha-SMA) actin isoforms and other muscle-typical proteins in the CS of human and rat hearts at different ages. SAN and IB cardiomyocytes were characterized by the presence of alpha-SMA, alpha-CA, calponin and caldesmon, whereas alpha-SKA and vimentin were absent. Double immunofluorescence demonstrated the co-localisation of alpha-SMA and alpha-CA in I-bands of SAN cardiomyocytes. AVN, AVB, BB and PF cardiomyocytes were alpha-SMA, calponin, caldesmon and vimentin negative, and alpha-CA and alpha-SKA positive. No substantial differences in actin isoform distribution were observed in human and rat hearts, except for the presence of isolated subendocardial alpha-SMA positive cardiomyocytes co-expressing alpha-CA in the ventricular septum of the rat. Aging did not influence CS cardiomyocyte actin isoform expression profile. These findings support the concept that cardiomyocytes of SAN retain the phenotype of a developing myogenic cell throughout the entire life span. PMID- 19281785 TI - TRAIL pathway components and their putative role in granulosa cell apoptosis in the human ovary. AB - Extensive apoptotic oocyte reduction occurs during fetal ovarian development. The regulatory pathways responsible for oocyte selection to programmed cell death are, however, poorly understood. The aim of this study was to investigate the potential involvement of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its death receptors TRAIL-R1/DR4 and TRAIL-R2/DR5 and decoy receptors TRAIL-R3/DcR1 and TRAIL-R4/DcR2 in the apoptotic process characterizing human fetal and adult ovaries. For this purpose, in situ hybridization and immunohistochemistry were applied to human fetal and adult ovarian samples to study the mRNA and protein expression of TRAIL pathway components, and a human granulosa cell tumor-derived cell line (KGN) was used to elucidate functional effects of TRAIL on apoptosis. TRAIL was expressed in human fetal ovary from the 11th week until term. The pro-apoptotic TRAIL-R2/DR5 and the anti-apoptotic TRAIL R4/DcR2 were also expressed in human ovaries throughout the fetal period. Among the different ovarian cell types, these TRAIL pathway components were mainly localized in the oocytes, and their expression increased towards term. Expression of TRAIL-R1/DR4 and TRAIL-R3/DcR1 was negligible in all of the fetal ovaries studied. Adult ovaries expressed TRAIL, TRAIL-R2/DR5, TRAIL-R3/DcR1 and TRAIL R4/DcR2 in granulosa cells and oocytes of small primary/secondary follicles as well as in granulosa and theca cells of more developed antral follicles. In KGN cells, TRAIL efficiently induced apoptosis in a dose-dependent manner, and this was blocked by a caspase inhibitor. The results indicate a role of the TRAIL pathway components in the regulation of granulosa cell apoptosis in in vitro and suggest that these factors may have a role in regulating ovarian apoptosis also in vivo. PMID- 19281786 TI - Postnatal expression of V2 vasopressin receptor splice variants in the rat cerebellum. AB - The V(2) vasopressin receptor gene contains an alternative splice site in exon-3, which leads to the generation of two splice variants (V(2a) and V(2b)) first identified in the kidney. The open reading frame of the alternatively spliced V(2b) transcript encodes a truncated receptor, showing the same amino acid sequence as the canonical V(2a) receptor up to the sixth transmembrane segment, but displaying a distinct sequence to the corresponding seventh transmembrane segment and C-terminal domain relative to the V(2a) receptor. Here, we demonstrate the postnatal expression of V(2a) and V(2b) variants in the rat cerebellum. Most importantly, we showed by in situ hybridization and immunocytochemistry that both V(2) splice variants were preferentially expressed in Purkinje cells, from early to late postnatal development. In addition, both variants were transiently expressed in the neuroblastic external granule cells and Bergmann fibers. These results indicate that the cellular distributions of both splice variants are developmentally regulated, and suggest that the transient expression of the V(2) receptor is involved in the mechanisms of cerebellar cytodifferentiation by AVP. Finally, transfected CHO-K1 expressing similar amounts of both V(2) splice variants, as that found in the cerebellum, showed a significant reduction in the surface expression of V(2a) receptors, suggesting that the differential expression of the V(2) splice variants regulates the vasopressin signaling in the cerebellum. PMID- 19281788 TI - Eupatilin exhibits a novel anti-tumor activity through the induction of cell cycle arrest and differentiation of gastric carcinoma AGS cells. AB - In many cases, the process of cancer cell differentiation is associated with the programmed cell death. In the present study, interestingly, we found that eupatilin, one of the pharmacologically active ingredients of Artemisia asiatica that has been reported to induce apoptosis in human gastric cancer AGS cells, also triggers differentiation of these cells. Treatment of AGS cells with eupatilin induced cell cycle arrest at the G(1) phase with the concomitant induction of p21(cip1), a cell cycle inhibitor. This led us to test whether eupatilin may trigger AGS cells to differentiate into the matured phenotypes of epithelial cells and this phenomenon may be coupled to the apoptosis. Eupatilin induced changes of AGS cells to a more flattened morphology with increased cell size, granularity, and mitochondrial mass. It also markedly induced trefoil factor 1 (TFF1), a gene responsible for the gastrointestinal cell differentiation. Eupatilin dramatically induced redistribution of tight junction proteins such as occludin and ZO-1, and F-actin at the junctional region between cells. It also induced phosphorylation of extracellular signal-regulated kinase 2 and p38 kinase. Blockade of ERK signaling by PD098059 or the dominant-negative ERK2 significantly reduced eupatilin-induced TFF1 and p21 expression as well as ZO-1 redistribution, indicating that ERK cascades may mediate eupatilin-induced AGS cell differentiation. Collectively, our results suggest that eupatilin acts as a novel anti-tumor agent by inducing differentiation of gastrointestinal cancer cells rather than its direct role in inducing apoptotic cell death. PMID- 19281787 TI - The chemokine SDF-1/CXCL12 regulates the migration of melanocyte progenitors in mouse hair follicles. AB - Mouse skin melanocytes originate from the neural crest and subsequently invade the epidermis and migrate into the hair follicles (HF) where they proliferate and differentiate. Here we demonstrate a role for the chemokine SDF-1/CXCL12 and its receptor CXCR4 in regulating the migration and positioning of melanoblasts during HF formation and cycling. CXCR4 expression by melanoblasts was upregulated during the anagen phase of the HF cycle. CXCR4-expressing cells in the HF also expressed the stem cell markers nestin and LEX, the neural crest marker SOX10 and the cell proliferation marker PCNA. SDF-1 was widely expressed along the path taken by migrating CXCR4-expressing cells in the outer root sheath (ORS), suggesting that SDF-1-mediated signaling might be required for the migration of CXCR4 cells. Skin sections from CXCR4-deficient mice, and skin explants treated with the CXCR4 antagonist AMD3100, contained melanoblasts abnormally concentrated in the epidermis, consistent with a defect in their migration. SDF-1 acted as a chemoattractant for FACS-sorted cells isolated from the anagen skin of CXCR4-EGFP transgenic mice in vitro, and AMD3100 inhibited the SDF-1-induced migratory response. Together, these data demonstrate an important role for SDF-1/CXCR4 signaling in directing the migration and positioning of melanoblasts in the HF. PMID- 19281789 TI - Inheritance of susceptibility to induction of nephroblastomas in the Noble rat. AB - Noble (Nb) strain rats are susceptible to nephroblastoma induction with transplacental exposure to direct-acting alkylating agent N-nitrosoethylurea (ENU), while F344 strain rats are highly resistant. To study the inheritance of susceptibility to induction of these embryonal renal tumors, fetal Nb and F344 rats and F1, F2 and reciprocal backcross hybrids were exposed transplacentally to ENU once on day 18 of gestation. Nephroblastomas developed in 53% of Nb offspring with no apparent gender difference, while no nephroblastomas developed in inbred F344 offspring. F1 and F2 hybrid offspring had intermediate responses, 28% and 30%, respectively. Nephroblastoma incidence in the offspring of F1 hybrids backcrossed to the susceptible strain Nb was 46%, while that in F1 hybrids backcrossed to resistant strain F344 was much lower (16%). Carcinogenic susceptibility is therefore consistent with the involvement of one major autosomal locus; the operation of a gene dosage effect; and a lack of simple Mendelian dominance for either susceptibility or resistance. Since established Wilms tumor-associated suppressor genes, Wt1 and Wtx, were not mutated in normal or neoplastic tissues, genomic profiling was performed on isolated Nb and F344 metanephric progenitors to identify possible predisposing factors to nephroblastoma induction. Genes preferentially elevated in expression in Nb rat progenitors included Wnt target genes Epidermal growth factor receptor, Inhibitor of DNA binding 2, and Jagged1, which were further increased in nephroblastomas. These studies demonstrate the value of this model for genetic analysis of nephroblastoma development and implicate both the Wnt and Notch pathways in its pathogenesis. PMID- 19281790 TI - An analytical system for the characterization of highly heterogeneous mixtures of N-linked oligosaccharides. AB - A novel system for characterizing complex N-linked oligosaccharide mixtures that uses a combination of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), capillary electrophoresis (CE), and high performance liquid chromatography (HPLC) has been developed. In this study, oligosaccharides released from recombinant TNK-tPA (tissue plasminogen activator) were derivatized with 5-amino-2-naphthalenesulfonic acid (ANSA). The negative charge imparted by the ANSA label facilitated the analysis of the oligosaccharides by MALDI-TOF MS by allowing the observation of both neutral and sialylated oligosaccharides in a single negative ion mode spectrum. Labeling with ANSA was also determined to be advantageous in the characterization of oligosaccharides by both HPLC and CE. The ANSA label was demonstrated to provide superior resolution over the commonly used label 8-aminopyrene-1,3,6-trisulfonic acid (APTS) in both the CE and HPLC analysis of oligosaccharides. To date, no other labels that enable the analysis of complex oligosaccharide mixtures in a single mass spectral mode, while also enabling high-resolution chromatographic and electrophoretic separation of the oligosaccharides, have been reported. By integrating the structural information obtained by MALDI-TOF MS analysis with the ability of CE and HPLC to discriminate between structural isomers, the complete characterization of complex oligosaccharide mixtures is possible. PMID- 19281791 TI - A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptides. AB - Fluorescence resonance energy transfer (FRET) is a distance-dependent interaction between the electronic excited states of two dye molecules. Here we introduce a novel FRET system for the detection of phosphopeptides using a phosphate-binding tag molecule, Zn(2+)-Phos-tag (1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2 olato dizinc(II) complex) attached with a 7-amino-4-methylcoumarin-3-acetic acid (AMCA). Carboxyfluorescein (FAM)-labeled phospho- and nonphosphopeptides were prepared as the target molecules for the FRET system. A set of FAM (a fluorescent acceptor, lambda(em) 520nm) and AMCA (a fluorescent donor, lambda(ex) 345nm) is frequently used for a FRET system. The AMCA-labeled Zn(2+)-Phos-tag specifically captured the FAM-labeled phosphopeptide to form a stable 1:1 complex, resulting in efficient FRET. After the FAM-labeled phosphopeptide was dephosphorylated with alkaline phosphatase, the FRET disappeared. Using this FRET system, we demonstrated the detection of the time-dependent dephosphorylation of the FAM labeled protein-tyrosine phosphatase 1B substrate. PMID- 19281792 TI - From protons to OXPHOS supercomplexes and Alzheimer's disease: structure-dynamics function relationships of energy-transducing membranes. AB - By the elucidation of high-resolution structures the view of the bioenergetic processes has become more precise. But in the face of these fundamental advances, many problems are still unresolved. We have examined a variety of aspects of energy-transducing membranes from large protein complexes down to the level of protons and functional relevant picosecond protein dynamics. Based on the central role of the ATP synthase for supplying the biological fuel ATP, one main emphasis was put on this protein complex from both chloroplast and mitochondria. In particular the stoichiometry of protons required for the synthesis of one ATP molecule and the supramolecular organisation of ATP synthases were examined. Since formation of supercomplexes also concerns other complexes of the respiratory chain, our work was directed to unravel this kind of organisation, e.g. of the OXPHOS supercomplex I(1)III(2)IV(1), in terms of structure and function. Not only the large protein complexes or supercomplexes work as key players for biological energy conversion, but also small components as quinones which facilitate the transfer of electrons and protons. Therefore, their location in the membrane profile was determined by neutron diffraction. Physico-chemical features of the path of protons from the generators of the electrochemical gradient to the ATP synthase, as well as of their interaction with the membrane surface, could be elucidated by time-resolved absorption spectroscopy in combination with optical pH indicators. Diseases such as Alzheimer's dementia (AD) are triggered by perturbation of membranes and bioenergetics as demonstrated by our neutron scattering studies. PMID- 19281793 TI - Mouse CD4+ CD25+ T regulatory cells are protected from autologous complement mediated injury by Crry and CD59. AB - Self cells depend on surface complement regulators to protect them from autologous complement mediated attack. CD4(+)CD25(+)foxp3(+) T regulatory (Treg) cells are critical in maintaining immune homeostasis, however, which complement regulators are expressed on them and how they are protected from autologous complement attack remains unknown. We report here that mouse Treg cells express virtually no DAF or CR1. Instead, all of them express Crry and approximately half of them express CD59. Both Crry(-/-) and CD59(-/-) Treg cells exhibit greater complement mediated injury than WT Treg cells. These results clarify the status of cell surface complement regulators on mouse Treg cells and indicate that both Crry and CD59 are required to protect Treg cells from autologous complement mediated injury. Additionally, these data also argue that different from previous assumption, at least in mice, CD4(+)CD25(+)foxp3(+) Treg cells are not homogenous and could be further divided into subgroups based on CD59 expression. PMID- 19281794 TI - PU.1, a novel caspase-3 substrate, partially contributes to chemotherapeutic agents-induced apoptosis in leukemic cells. AB - PU.1 is one of key regulators of hematopoietic cell development, a tightly regulated lineage-specific process. Here we provide the first evidence that PU.1 protein is cleaved into two fragments of 24 kDa and 16 kDa during apoptosis progression in leukemic cell lines and primary leukemic cells. Further experiments with specific capase-3 inhibitor Z-DEVD-fmk and the in vitro proteolytic system confirmed that PU.1 is a direct target of caspase-3. Using site-directed mutagenesis analyses, the aspartic acid residues at positions 97 and 151 of PU.1 protein were identified as capsase-3 target sites. More intriguingly, the suppression of PU.1 expression by small interfering RNAs (siRNAs) significantly inhibits DNA-damaging agents NSC606985 and etoposide induced apoptosis in leukemic cells, together with the up-regulated expression of anti-apoptotic bcl-2 gene. These results would provide new insights for understanding the mechanism of PU.1 protein in hematopoiesis and leukemogenesis. PMID- 19281795 TI - A Jak2 inhibitor, AG490, reverses lipin-1 suppression by TNF-alpha in 3T3-L1 adipocytes. AB - Lipin-1 is a multifunctional metabolic regulator, involving in triacylglycerol and bioactive glycerolipids synthesis as an enzyme, transcriptional regulation as a coactivator, and adipogenesis. In obesity, adipose lipin-1 expression is decreased. Although lipin-1 is implicated in the pathogenesis of obesity, the mechanism is still not clear. Since TNF-alpha is deeply involved in the pathogenesis of obesity, insulin resistance, and diabetes, here we investigated the role of TNF-alpha on lipin-1 expression in adipocytes. Quantitative PCR studies showed that TNF-alpha suppressed both lipin-1A and -1B isoform expression in time- and dose-dependent manners in mature 3T3-L1 adpocytes. A Jak2 inhibitor, AG490, reversed the suppressive effect of TNF-alpha on both lipin-1A and -1B. In contrast, NF-kappaB, MAPKs, ceramide, and beta-catenin pathway tested were not involved in the mechanism. These results suggest that TNF-alpha could be involved in obesity-induced lipin-1 suppression in adipocytes and Jak2 may play an important role in the mechanism. PMID- 19281796 TI - Regulation of FOXO1-mediated transcription and cell proliferation by PARP-1. AB - Forkhead box O (FOXO) transcription factors play an important role in a wide range of biological processes, including cell cycle control, apoptosis, detoxification of reactive oxygen species, and gluconeogenesis through regulation of gene expression. In this study, we demonstrated that PARP-1 functions as a negative regulator of FOXO1. We showed that PARP-1 directly binds to and poly(ADP ribosyl)ates FOXO1 protein. PARP-1 represses FOXO1-mediated expression of cell cycle inhibitor p27(Kip1) gene. Notably, poly(ADP-ribosyl)ation activity was not required for the repressive effect of PARP-1 on FOXO1 function. Furthermore, knockdown of PARP-1 led to a decrease in cell proliferation in a manner dependent on FOXO1 function. Chromatin immunoprecipitation experiments confirmed that PARP 1 is recruited to the p27(Kip1) gene promoter through a binding to FOXO1. These results suggest that PARP-1 acts as a corepressor for FOXO1, which could play an important role in proper cell proliferation by regulating p27(Kip1) gene expression. PMID- 19281797 TI - Feasibility and efficacy of bone tissue engineering using human bone marrow stromal cells cultivated in serum-free conditions. AB - Current standard techniques for bone tissue engineering utilize ex vivo expanded osteogenic cells. However, ex vivo expansion requires serum, which may hinder clinical applications. Here, we report the feasibility and efficacy of bone tissue engineering with human bone marrow stromal cells (BMSCs) expanded in serum free conditions. Bone marrow was aspirated from 4 healthy donors and adherent cells were cultured in either serum-free medium (STEMPRO((R)) MSC SFM) or conventional serum-containing medium (alpha-MEM supplemented with 10% serum). Efficacy of expansion was greater in serum-free medium. Phenotypically, serum free expanded BMSCs were smaller in cell-size and showed expression of CD105(++) and CD146(dim). After osteogenic induction, serum-free expanded BMSCs showed lower alkaline phosphatase activity. However, they showed higher responsiveness to induction. In vivo bone-forming ability was also confirmed. In conclusion, bone tissue engineering with serum-free expanded BMSCs is feasible and as efficient as that obtained with BMSCs expanded in conventional serum-containing medium. PMID- 19281798 TI - Cleavage of chemokines CCL2 and CXCL10 by matrix metalloproteinases-2 and -9: implications for chemotaxis. AB - Proteolytic processing of chemokines is a complex process that can result in dramatic effects on their chemotactic activity. Results from gel electrophoresis and mass spectrometry using recombinant CCL2 and CXCL10, incubated with either MMP-2 or -9, indicate that both chemokines are cleaved by the enzymes. N-terminal truncation of four amino acids from CCL2, and four or five residues from CXCL10 occurred, but removal of four residues from the C-terminus of CXCL10 was also observed with both MMPs. The speed of the reaction was chemokine-dependent, with N-terminal processing of CCL2 being complete within 3h, whereas activity of the MMPs on CXCL10 remained incomplete at 48h. The effect on the chemotactic potential of N-terminal truncation of CCL2 by MMPs-2 and -9 was investigated using in vitro migration assays. Monocytic cells exhibited a 2-fold reduction in migration to MMP-cleaved CCL2 variants, compared to intact CCL2. PMID- 19281799 TI - Divergent effects of estradiol and the estrogen receptor-alpha agonist PPT on eating and activation of PVN CRH neurons in ovariectomized rats and mice. AB - Eating is modulated by estradiol in females of many species and in women. To further investigate the estrogen receptor mechanism mediating this effect, ovariectomized rats and mice were treated with estradiol benzoate or the estrogen receptor-alpha (ER-alpha)-selective agonist PPT. PPT inhibited eating in rats much more rapidly than estradiol (approximately 2-6 h versus >24 h). In contrast, the latencies to vaginal estrus after PPT and estradiol were similar (>24 h). PPT also inhibited eating within a few hours in wild-type mice, but failed to inhibit eating in transgenic mice deficient in ER-alpha (ERalphaKO mice). PPT, but not estradiol, induced the expression of c-Fos in corticotrophin-releasing hormone (CRH)-expressing cells of the paraventricular nucleus (PVN) of the hypothalamus within 90-180 min in rats. Both PPT and estradiol reduced c-Fos expression in an ER-alpha-containing area of the nucleus of the solitary tract. The anomalously rapid eating-inhibitory effect of PPT suggests that PPT's neuropharmacological effect differs from estradiol's, perhaps because PPT differentially activates membrane versus nuclear ER-alpha or because PPT activates non-ER-alpha membrane estrogen receptors in addition to ER-alpha. The failure of PPT to inhibit eating in ERalphaKO mice, however, indicates that ER-alpha is necessary for PPT's eating inhibitory action and that any PPT-induced activation of non-ER-alpha estrogen receptors is not sufficient to inhibit eating. Finally, the rapid induction of c Fos in CRH-expressing cells in the PVN by PPT suggests that PPT elicits a neural response that is similar to that elicited by stress or aversive emotional stimuli. PMID- 19281800 TI - The precerebellar linear nucleus in the mouse defined by connections, immunohistochemistry, and gene expression. AB - The linear nucleus (Li) is a prominent cell group in the caudal hindbrain, which was first described in a study of cerebellar afferents in the rat by [Watson, C.R.R., Switzer, R.C. III, 1978. Trigeminal projections to cerebellar tactile areas in the rat origin mainly from N. interpolaris and N. principalis. Neurosci. Lett. 10, 77-82.]. It was named for its elongated appearance in transverse sections. Since this original description in the rat, reference to the nucleus seems to have been largely absent from experimental studies of mammalian precerebellar nuclei. We therefore set out to define the cytoarchitecture, cerebellar connections, and molecular characteristics of Li in the mouse. In coronal Nissl sections at the level of the rostral inferior olive, it consists of two parallel bands of cells joined at their dorsal apex by a further band of cells, making the shape of the Greek capital letter pi. Our three-dimensional reconstruction demonstrated that the nucleus is continuous with the lateral reticular nucleus (LRt) and that the ambiguus nucleus sits inside the arch of Li. Cerebellar horseradish peroxidase injections confirmed that the cells of Li project to cerebellum. We have shown that Li cells express Atoh1 and Wnt1 lineage markers that are known to label the rhombic lip derived precerebellar nuclei. We have examined the relationship of Li cells to a number of molecular markers, and have found that many of the cells express a nonphosphorylated epitope in neurofilament H (SMI 32), a feature they share with the LRt. The mouse Li therefore appears to be a rostrodorsal extension of the LRt. PMID- 19281801 TI - A lack of default network suppression is linked to increased distractibility in ADHD. AB - Heightened distractibility in participants with ADHD as indexed by increased reaction time (RT) variability has been hypothesized to be due to a failure to sufficiently suppress activation in the default attention network during cognitively demanding situations. The present study utilized fMRI to examine the relationship between intra-individual variability (IIV) in task RT and suppression of BOLD response in regions of the default network, using a working memory paradigm and two levels of control tasks. IIV was calculated separately for thirteen healthy control and twelve children with ADHD, Combined Type. Children with ADHD displayed significantly more RT variability than controls. Neural measures showed that although both groups displayed a pattern of increasing deactivation of the medial prefrontal cortex (PFC) with increasing task difficulty, the ADHD group was significantly less deactive than controls. Correlations between IIV and brain activation suggested that greater variability was associated with a failure to deactivate ventromedial PFC with increasing task difficulty. T-tests on brain activation between participants with ADHD with low versus high IIV implicated a similar region so that high variability was associated with greater activity in this region. These data provide support for the theory that increased distractibility in at least some participants with ADHD may be due to an inability to sufficiently suppress activity in the default attention network in response to increasing task difficulty. PMID- 19281802 TI - Influence of the 5-HTTLPR polymorphism and environmental risk factors in a Brazilian sample of patients with autism spectrum disorders. AB - The 5-HTTLPR polymorphism of serotonin transporter gene is widely investigated in association studies in autism spectrum disorders (ASD). The results of such studies, however, remain controversial possibly due to the great genetic heterogeneity related to ASD and the lack of evaluation of the triallelic functional structure of 5-HTTLPR. This study tested for association between the 5 HTTLPR and ASD in a Brazilian sample by case-control and family-based association test (FBAT) methods, considering the biallelic and triallelic structures of this polymorphism. In addition, we performed an exploratory analysis of associations between specific clinical outcomes of ASD patients and 5-HTTLPR as well as several prenatal environmental factors. Genotyping was achieved in 151 ASD patients, 179 unrelated controls and 105 complete trios. There was no evidence of association between the 5-HTTLPR with ASD in both case-control and FBAT tests, but the LaLa 5-HTTLPR genotype was associated with mood instability in patients (P=0.006). The prenatal exposure to potential neuroteratogenic drugs was associated with epilepsy (P<0.001). Our findings suggest that the 5-HTTLPR is not associated with ASD in the Brazilian population, even considering the triallelic structure. Additionally, this study suggested a role of the 5-HTTLPR and environmental factors in the clinical expression of ASD. PMID- 19281803 TI - A family-based study of the IL3RA gene on susceptibility to schizophrenia in a Chinese Han population. AB - Schizophrenia has been observed to be associated with various abnormalities in cytokines and cytokine receptors that have been one of the recent focal points of immunological research in schizophrenia. Recent reports have showed that IL-3 gene, colony stimulating factor 2 receptor alpha (CSF2RA) and IL-3 receptor alpha (IL3RA) are associated with schizophrenia. The aim of this study was to investigate IL3RA gene variants in schizophrenia among a Chinese population by using a family-based association approach. Our sample included 101 Chinese parent offspring trios of Han descent. All subjects were genotype for IL3RA-rs6603272 and -rs6645249 using PCR techniques. Single marker analysis showed a significant association for rs6603272 (X(2)=5.15, df=1, P=0.023), but not for the rs6645249. However, there was a significant genotypic association of both the polymorphisms with schizophrenia (for rs6603272, X(2)=6.15, df=2, P=0.046; for rs6645249, X(2)=21.79, df=2, P=1.85e-005). Haplotype TDT was statistically significant (X(2)=5.14, df=1, P=0.023), with the rs6603272(T)-rs6645249(G) haplotype significantly associated with schizophrenia (OR=1.66; 95% CI=1.08-2.55). In conclusion, our family-based association study also revealed a small but significant contribution of the IL3RA variants to susceptibility to schizophrenia in a Chinese population. PMID- 19281804 TI - Optimizing the yield and utility of circulating cell-free DNA from plasma and serum. AB - BACKGROUND: Cell-free DNA (CFDNA) in the plasma/serum of patients with cancer demonstrates tumour-associated genetic alterations, offering possibilities for diagnosis, prognostication and disease monitoring. There is wide variation in the reported levels of CFDNA, associated with different methods used to collect, process and analyze blood samples. We therefore evaluated different aspects of laboratory protocols for the processing and purification of CFDNA in clinical studies. METHODS: We evaluated and compared the QIAamp kit and a Triton/Heat/Phenol protocol (THP) for CFDNA purification. Total CFDNA was quantified by PicoGreen assay and SYBR-Green real-time PCR assay was used to amplify specific genes to estimate the efficiency of different protocols. RESULTS: The efficiency of DNA extraction was 18.6% using the standard QIAamp protocol and 38.7% using the THP method (p < 0.0001, unpaired t-test). A modified QIAamp protocol that included a proteinase incubation stage and elution volumes up to 300 microl increased DNA yields, but was not as good as the THP method. CONCLUSIONS: Blood samples should be kept at/or below room temperature (18 degrees C-22 degrees C) for no more than 2 h before plasma separation by double spin. Because of its higher efficiency, low-cost and good-quality products, the THP protocol is preferred for extraction of CFDNA. PMID- 19281805 TI - Evaluation of serum percent trisialotransferrin as potential predictive biomarker of hepatocellular dedifferentiation in chronic liver disease. AB - BACKGROUND AND AIM: Chronic hepatitis induced liver fibrogenesis is characterized by epithelial-to-mesenchymal transition of liver parenchymal cells (hepatocytes) to fibroblast (-like cells), i.e. increasing hepatocellular dedifferentiation, ultimatively leading to the development of hepatocellular carcinoma (HCC). Up to now the spectrum of valid serum biomarkers for this process of hepatocellular dedifferentiation is very limited. We therefore investigated the dynamics of alterations in the serum transferrin isoform pattern in the pathogenetic sequence from liver fibrosis to hepatocellular carcinoma, to evaluate the suitability of one of the isoforms as potential biomarker for hepatocellular dedifferentiation in chronic liver disease. RESULTS: Our data on 252 patients with hepatitis C virus (HCV) induced fibrogenic liver disease and on 43 patients with HCV induced HCC demonstrate a dynamic alteration of serum % trisialotransferrin levels in the pathogenetic sequence from early stage hepatic fibrosis to fully developed hepatocellular carcinoma, whereas serum % di- and pentasialotransferrin values seem not to be affected. We show that patients with early stage fibrosis (METAVIR stage F1) and weak fibrogenic activitiy (METAVIR grade A1) display significantly lower values of serum % trisialotransferrin compared to healthy controls, and that serum % trisialotransferrin values increased steadily parallel to an increase of fibrotic stage and grade, respectively, while finally exceeding normal values in those patients with hepatocellular carcinoma. CONCLUSION: These findings propose a possible diagnostic value of serum % trisialotransferrin concentrations in the pathogenesis of hepatocellular dedifferentiation and the use of this parameter as possible predictive tumor marker in patients with chronic liver disease. Monitoring the pattern of transferrin bound sialic acid residues may thus be a helpful tool in assessing the risk of malignant degeneration in patients with chronic fibrogenic liver disease. PMID- 19281806 TI - Altered amino acid homeostasis in subjects affected by fibromyalgia. AB - OBJECTIVES: To evaluate plasma amino acid (AA) concentrations in patients affected by fibromyalgia (FM) and to study the relationships between their levels and FM clinical parameters. DESIGN AND METHODS: 20 AAs were assessed in 34 FM patients and in 18 healthy volunteers by means of a modified version of the Waters picotag method. RESULTS: Significant lower plasma taurine, alanine, tyrosine (Tyr), valine, methionine, phenylalanine and threonine concentrations, and the sum of essential AAs were observed in FM patients vs healthy controls (P<0.05). Tyr CAA' ratio and the sum of AAs competing with tryptophan for brain uptake were significantly reduced in FM (P<0.05). A significant correlation was found between FM clinical parameters and certain AAs. CONCLUSIONS: Our results suggest probable defects of gut malabsorption of certain AAs in FM patients. Moreover, given the reduced Tyr CAA' ratio in FM patients, a possible impairment of the cathecolaminergic system in the FM syndrome may be suggested. PMID- 19281807 TI - Performance evaluation of a particle-enhanced turbidimetric cystatin C assay on the Abbott ci8200 analyzer. AB - OBJECTIVES: Glomerular filtration rate (GFR) is widely accepted as the best overall measure of kidney function. Cystatin C is a novel endogenous GFR marker that has been shown to be superior to creatinine for estimation of GFR in several studies. There is a need for cystatin C assays adapted to routine chemistry instrument to minimize turnaround times and allowing 24 h/day availability. MATERIALS AND METHODS: We have evaluated a new cystatin C assay developed for Architect cSystem (Abbott Laboratories, Abbott Park, IL, USA). RESULTS: The cystatin C assay showed good agreement with the corresponding assay from Dade Behring (Deerfield, IL, USA). The assay has a very low total imprecision and a good linearity. CONCLUSIONS: The new cystatin C assay is an interesting alternative to current cystatin C assays. On an Architect cSystem the assay can be performed with the same turnaround times and availability as creatinine. PMID- 19281808 TI - Involvement of glucocorticoid receptor and peroxisome proliferator activated receptor-gamma in pioglitazone mediated chronic gastric ulcer healing in rats. AB - Evidences suggest Peroxisome Proliferator Activated Receptor-gamma (PPAR-gamma) ligand, pioglitazone results in the attenuation of gastric mucosal injury. But the molecular mechanism through which these agonists actually elicit gastroprotection through modulating inflammatory responses has not yet been established. Chronic gastric ulcer induced in rats by intraluminal application of acetic acid resulted in elevation of proinflammatory cytokines gene expression, such as, TNF-alpha (Tumor Necrosis Factor-alpha), IL-1beta (Interleukin-1beta) and the protein levels of nuclear p65 subunit of NF-kappaB (Nuclear Factor kappaB) but decreased levels of PPAR-gamma gene expression. Pioglitazone treatment reduced the severity of ulceration, repressed levels of TNF-alpha, IL 1beta and nuclear p65 subunit as well as increased the abundance of PPAR-gamma in gastric mucosa. Moreover, it significantly upregulated protein levels of glucocorticoid receptor demonstrating its possible involvement in pioglitazone mediated ulcer healing along with PPAR-gamma. Administration of pioglitazone reverted back the decreased levels of both PPAR-gamma and glucocorticoid receptor, resulting in their redistribution to the nucleus from the cytosol in course of ulcer healing. Moreover, pharmacological inhibition of glucocorticoid receptor function by its antagonist (RU486) inhibited pioglitazone mediated downregulation of TNF-alpha and IL-1beta gene expression confirming involvement of glucocorticoid receptor in pioglitazone mediated ulcer healing. Co immunoprecipitation studies further established association of PPAR-gamma with glucocorticoid receptor during ulcer healing which was enhanced following pioglitazone administration. Thus, the present study is first of its kind bearing direct relevance to the participation of both PPAR-gamma and glucocorticoid receptor and their physical association in influencing amelioration of inflammatory responses during pioglitazone mediated gastric ulcer healing. PMID- 19281809 TI - Sergliflozin etabonate, a selective SGLT2 inhibitor, improves glycemic control in streptozotocin-induced diabetic rats and Zucker fatty rats. AB - The low-affinity sodium glucose cotransporter (SGLT2) is responsible for most of the glucose reabsorption in the kidney and has been highlighted as a novel therapeutic target for the treatment of diabetes. We discovered sergliflozin etabonate, a novel selective SGLT2 inhibitor, and found that selective inhibition of SGLT2 increased urinary glucose excretion and consequently decreased plasma glucose levels. In this report, we examined the antihyperglycemic effects of sergliflozin etabonate in normal and diabetic rats in comparison with those of a sulfonylurea (gliclazide) and an alpha-glucosidase inhibitor (voglibose). Sergliflozin etabonate increased urinary glucose excretion in a dose-dependent manner, and inhibited the increase in plasma glucose after sucrose loading independently of insulin secretion in normal rats. Sergliflozin etabonate also improved postprandial hyperglycemia in neonatal streptozotocin-induced diabetic rats; whereas gliclazide did not improve it. In rats with mild or moderate streptozotocin-induced diabetes, the degree of the antihyperglycemic effects of sergliflozin etabonate correlated with the severity of the diabetic condition. Sergliflozin etabonate did not affect the plasma glucose level of normal rats as seen with gliclazide. Chronic treatment with sergliflozin etabonate reduced the levels of glycated hemoglobin and fasting plasma glucose, and improved the glycemic response after glucose loading in Zucker fatty rats. In addition, sergliflozin etabonate did not affect the body weight or food intake. These data indicate that sergliflozin etabonate could improve glycemic control without its use resulting in insulin secretion, hypoglycemia, and body weight gain, and may provide a unique approach to the treatment of diabetes. PMID- 19281810 TI - Angiotensin II-induced hypertension regulates AT1 receptor subtypes and extracellular matrix turnover in mouse retinal pigment epithelium. AB - Accumulation of specific deposits and extracellular molecules under the retinal pigment epithelium (RPE) has been previously observed in eyes with age-related macular degeneration (AMD) and may play a role in the pathogenesis of AMD. Even though age is the major determinant for developing AMD, clinical studies have revealed hypertension (HTN) as another systemic risk factor. Angiotensin II (Ang II) is considered the most important hormone associated with HTN. To evaluate the relationship of Ang II to AMD, we studied whether mouse RPE expresses functional Ang II receptor subtypes and whether HTN-induced Ang II regulates expression of these receptors as well as critical ECM molecules (MMP-2 and type IV collagen) involved in ECM turnover in RPE. We used 9-month-old C57BL/6 male mice infused with Ang II alone or Ang II in combination with the AT1 receptor antagonist candesartan or the AT2 receptor antagonist PD123319 for 4 weeks to determine whether HTN-associated Ang II was important for ECM regulation in RPE. We found that mouse RPE expressed both Ang II receptor subtypes at the mRNA and protein levels. Infusion with Ang II induced HTN and elevated plasma and ocular Ang II levels. Ang II also regulated AT1a and AT1b receptor mRNA expression, the intracellular concentration of calcium [Ca(2+)](i), MMP-2 activity, and type IV collagen accumulation. Concurrent administration of Ang II with the AT1 receptor blocker prevented the increase in blood pressure and rise in ocular Ang II levels, as well as the calcium and MMP-2 responses. In contrast, the type IV collagen response to Ang II was prevented by blockade of AT2 receptors, but not AT1 receptors. Plasma Ang II levels were not modified by the AT1 or AT2 receptor blockade. Since the effects of Ang II on MMP-2 and type IV collagen require inhibition of both Ang II receptor subtypes, these receptors may play a role as a potential therapeutic targets to prevent ECM turnover dysregulation in the RPE basement membrane, suggesting a pathogenic mechanism to explain the link between HTN and AMD. PMID- 19281811 TI - Acute corticosterone elevation enhances antipredator behaviors in male tree lizard morphs. AB - In response to stressful events, most vertebrates rapidly elevate plasma glucocorticoid levels. Corticosterone release stimulates physiological and behavioral responses that can promote survival while suppressing behaviors that are not crucial to immediate survival. Corticosterone also has preparatory effects for subsequent stressors. Using male tree lizards (Urosaurus ornatus), we tested our prediction that elevated corticosterone is important for mediating and enhancing antipredator behaviors. Male tree lizards express developmentally fixed polymorphisms that are mediated by early organizational actions of steroid hormones, and thus we also tested the hypothesis that morph-specific differences in antipredator behaviors of adults are independent of circulating corticosterone levels. Plasma corticosterone levels were elevated exogenously for 12-16 h using non-invasive dermal patches, and we then compared the behavioral responses of these corticosterone-patched males to control-patched males during a simulated encounter with a caged predator (collared lizard, Crotaphytus nebrius) in outdoor enclosures. Elevating corticosterone did not alter the antipredator behavioral repertoire of each male morph, but did enhance their responses during the predator encounter: all corticosterone-patched males responded more quickly, hid longer, and displayed more toward the predator than control-patched males. With the corticosterone patch, the non-territorial and wary orange morph was still behaviorally the most wary morph, responding more quickly and hiding longer than either the bolder orange-blue or mottled morphs. Smaller males were generally warier than larger males, regardless of the endocrine treatment or color morph type. In sum, elevated circulating corticosterone enhances antipredator responses for all male tree lizard morphs, without altering morph-specific or size-specific differences in their behavioral responses. PMID- 19281812 TI - Hippocampal damage abolishes the cortisol response to psychosocial stress in humans. AB - The hippocampus (HC) is necessary for learning and memory, but it also plays a role in other behaviors such as those related to stress and anxiety. In support of the latter idea, we show here that bilateral HC damage abolishes the cortisol response to psychosocial stress. We collected salivary cortisol, heart rate, and affective responses to the Trier Social Stress Test (TSST) from 7 participants with bilateral HC lesions, 12 participants with damage outside the HC, and 28 healthy normal comparison participants matched to the HC participants on age and sex. HC participants showed elevated pre-stress cortisol, but no cortisol response to the TSST. Heart rate and affective responses in the HC group were similar to those of the comparison groups. Participants with brain damage outside the HC showed stress responses that were comparable to those of the healthy comparison group. These findings support the idea that the functions of the human HC extend beyond learning and memory, and suggest that the HC is necessary for producing the cortisol response to psychosocial stress. PMID- 19281813 TI - Lab and field experiments: are they the same animal? AB - To advance our understanding of biological processes we often plan our experiments based on published data. This can be confusing though, as data from experiments performed in a laboratory environment are sometimes different from, or completely opposite to, findings from similar experiments performed in the "real world". In this mini-review, we discuss instances where results from laboratory experiments differ as a result of laboratory housing conditions, and where they differ from results gathered in the field environment. Experiments involving endocrinology and behavior appear to be particularly susceptible to influence from the environment in which they are performed. As such, we have attempted to promote discussion of the influence of housing environment on the reproductive axis, circadian biology and behavior, immune function, stress biology, neuroplasticity and photoperiodism. For example, why should a rodent species be diurnal in one housing environment yet nocturnal in another? Are data that are gathered from experiments in the laboratory applicable to the field environment, and vice-versa? We hope not only to highlight the need for experiments in both lab and field when looking at complex biological systems, but also to promote frank discussion of discordant data. Perhaps, just as study of individual variation has been gaining momentum in recent years, data from variation between experimental arenas can provide us with novel lines of research. PMID- 19281814 TI - Redox signalling and miRNA function in cardiomyocytes. PMID- 19281815 TI - Ventricular K+ currents are reduced in mice with elevated levels of serum TNFalpha. AB - In the present study mice were treated with tumor necrosis factor alpha (TNFalpha) for 6 weeks to determine if chronic TNFalpha treatment could produce serum levels of TNFalpha similar to what has been observed in disease states (heart failure, HIV) and to determine if these levels of TNFalpha alter ventricular K(+) currents. Mice chronically treated with TNFalpha and sham treated mice were utilized for experiments. Serum levels were measured with a Searchlight protein array. Patch-clamp techniques, real-time PCR and Western blot analysis were used to study K(+) current densities and K(+) channel expression. Results showed that serum concentrations of TNFalpha were significantly higher in TNFalpha treated mice compared to controls (control: 9.5+/-1.5 pg/ml, TNFalpha: 27.4+/-5.0 pg/ml; p<0.05) and comparable to serum TNFalpha levels observed in heart failure and HIV models. In ventricular myocytes from TNFalpha treated mice the outward K(+) currents I(to) and I(Kur) were significantly reduced (at +30 mV: I(to): control: 45.0+/-2.9 pA/pF, TNFalpha: 34.5+/-2.9 pA/pF; p<0.05; I(Kur): control 34.1+/-2.7 pA/pF, TNFalpha: 25.0+/-2.2 pA/pF; p<0.05). Expression studies revealed that ventricular mRNA and protein expression for the channels underlying I(to) and I(Kur) did not differ between the two groups. However, the recovery from inactivation for I(Kur) was significantly longer in TNFalpha treated mice. Overall, this study shows that pathologically relevant levels of serum TNFalpha modulate K(+) currents in mouse ventricle. These findings could help to explain the role of TNFalpha in the pathogenesis of cardiac arrhythmia. PMID- 19281817 TI - Metabolic pathfinding using RPAIR annotation. AB - Metabolic databases contain information about thousands of small molecules and reactions, which can be represented as networks. In the context of metabolic reconstruction, pathways can be inferred by searching optimal paths in such networks. A recurrent problem is the presence of pool metabolites (e.g., water, energy carriers, and cofactors), which are connected to hundreds of reactions, thus establishing irrelevant shortcuts between nodes of the network. One solution to this problem relies on weighted networks to penalize highly connected compounds. A more refined solution takes the chemical structure of reactants into account in order to differentiate between side and main compounds of a reaction. Thanks to an intensive annotation effort at KEGG, decompositions of reactions into reactant pairs (RPAIR) categorized by their role (main, trans, cofac, ligase, and leave) are now available. The goal of this article is to evaluate the impact of RPAIR data on pathfinding in metabolic networks. To this end, we measure the impact of different parameters concerning the construction of the metabolic network: mapping of reactions and reactant pairs onto a graph, use of selected categories of reactant pairs, weighting schemes for compounds and reactions, removal of highly connected metabolites, and reaction directionality. In total, we tested 104 combinations of parameters and identified their optimal values for pathfinding on the basis of 55 reference pathways from three organisms. The best-performing metabolic network combines the biochemical knowledge encoded by KEGG RPAIR with a weighting scheme penalizing highly connected compounds. With this network, we could recover reference pathways from Escherichia coli with an average accuracy of 93% (32 pathways), from Saccharomyces cerevisiae with an average accuracy of 66% (11 pathways), and from humans with an average accuracy of 70% (12 pathways). Our pathfinding approach is available as part of the Network Analysis Tools. PMID- 19281816 TI - Morphological dynamics of mitochondria--a special emphasis on cardiac muscle cells. AB - Mitochondria play a critical role in cellular energy metabolism, Ca(2+) homeostasis, reactive oxygen species generation, apoptosis, aging, and development. Many recent publications have shown that a continuous balance of fusion and fission of these organelles is important in maintaining their proper function. Therefore, there is a steep correlation between the form and function of mitochondria. Many major proteins involved in mitochondrial fusion and fission have been identified in different cell types, including heart. However, the functional role of mitochondrial dynamics in the heart remains, for the most part, unexplored. In this review we will cover the recent field of mitochondrial dynamics and its physiological and pathological implications, with a particular emphasis on the experimental and theoretical basis of mitochondrial dynamics in the heart. PMID- 19281818 TI - An efficient strategy for the determination of the three-dimensional architecture of ribonucleoprotein complexes by the combination of a few easily accessible NMR and biochemical data: intermolecular recognition in a U4 spliceosomal complex. AB - Ribonucleoprotein (RNP) complexes are involved in several cellular processes, including RNA processing, transcription and translation. RNP structures are often dynamic in nature, undergoing significant remodeling during the course of their function. Visualization of the three-dimensional arrangement of single components in the complex and characterization of the intermolecular interactions are essential for understanding the mechanisms of operation. Crystallization either is not always achievable for these highly dynamic RNP particles or requires trimming the complex to a stable, well-structured core that lacks the flexible, regulatory domains. Alternative techniques that can provide structural information for complexes in solution under native conditions, where they retain their natural dynamic properties, are needed. In this study, we explored the possibility of using a combination of NMR, biochemical data and molecular modeling to generate an accurate high-resolution model of RNP complexes. We applied this strategy to the ternary hPrp31 (human Prp31)-15.5K-U4 5'-SL (stem loop) spliceosomal complex, which, due to its large size and instability and because of the difficulty in obtaining isotopically labeled hPrp31, is not amenable to complete structure determination by NMR. We designed a protocol where the protein-protein interaction surface is defined for 15.5K by NMR data, while the relative orientations of the U4 RNA and the hPrp31 protein are described by mutational and cross-linking data. Using these data in a restrained ensemble docking protocol, we obtained a model for the ternary complex that reveals a novel rationale for the hierarchical assembly of the complex. Comparison of the docking model with the crystal structure recently obtained for a trimmed version of the complex reveals the high accuracy of the docking model, even down to an atomic level. This work shows that the architecture of large RNP complexes is within reach by NMR investigation in solution even for those cases where a traditional structural determination cannot be performed. PMID- 19281819 TI - Solution and crystal structures of mRNA exporter Dbp5p and its interaction with nucleotides. AB - DEAD-box protein 5 (Dbp5p) plays very important roles in RNA metabolism from transcription, to translation, to RNA decay. It is an RNA helicase and functions as an essential RNA export factor from nucleus. Here, we report the solution NMR structures of the N- and C-terminal domains (NTD and CTD, respectively) of Dbp5p from Saccharomyces cerevisiae (ScDbp5p) and X-ray crystal structure of Dbp5p from Schizosaccharomyces pombe (SpDbp5p) in the absence of nucleotides and RNA. The crystal structure clearly shows that SpDbp5p comprises two RecA-like domains that do not interact with each other. NMR results show that the N-terminal flanking region of ScDpbp5 (M1-E70) is intrinsically unstructured and the region Y71-R121 including the Q motif is highly dynamic on millisecond-microsecond timescales in solution. The C-terminal flanking region of ScDbp5p forms a short beta-strand and a long helix. This helix is unique for ScDbp5p and has not been observed in other DEAD-box proteins. Compared with other DEAD-box proteins, Dbp5p has an extra insert with six residues in the CTD. NMR structure reveals that the insert is located in a solvent-exposed loop capable of interacting with other proteins. ATP and ADP titration experiments show that both ADP and ATP bind to the consensus binding site in the NTD of ScDbp5p but do not interact with the CTD at all. Binding of ATP or ADP to NTD induces significant conformational rearrangement too. PMID- 19281820 TI - Desmin and vimentin intermediate filament networks: their viscoelastic properties investigated by mechanical rheometry. AB - We have investigated the viscoelastic properties of the cytoplasmic intermediate filament (IF) proteins desmin and vimentin. Mechanical measurements were supported by time-dependent electron microscopy studies of the assembly process under similar conditions. Network formation starts within 2 min, but it takes more than 30 min until equilibrium mechanical network strength is reached. Filament bundling is more pronounced for desmin than for vimentin. Desmin filaments (persistence length l(p) approximately 900 nm) are stiffer than vimentin filaments (l(p) approximately 400 nm), but both IFs are much more flexible than microfilaments. The concentration dependence of the plateau modulus G(0) approximately c(alpha) is much weaker than predicted theoretically for networks of semiflexible filaments. This is more pronounced for vimentin (alpha=0.47) than for desmin (alpha=0.70). Both networks exhibit strain stiffening at large shear deformations. At the transition from linear to nonlinear viscoelastic response, only desmin shows characteristics of nonaffine network deformation. Strain stiffening and the maximum modulus occur at strain amplitudes about an order of magnitude larger than those for microfilaments. This is probably attributable to axial slippage within the tetramer building blocks of the IFs. Network deformation beyond a critical strain gamma(max) results in irreversible damage. Strain stiffening sets in at lower concentrations, is more pronounced, and is less sensitive to ionic strength for desmin than for vimentin. Hence, desmin exhibits strain stiffening even at low-salt concentrations, which is not observed for vimentin, and we conclude that the strength of electrostatic repulsion compared to the strength of attractive interactions forming the network junctions is significantly weaker for desmin than for vimentin filaments. These findings indicate that both IFs exhibit distinct mechanical properties that are adapted to their respective cellular surroundings [i.e., myocytes (desmin) and fibroblasts (vimentin)]. PMID- 19281821 TI - PAC-1 activates procaspase-3 in vitro through relief of zinc-mediated inhibition. AB - The direct induction of apoptosis has emerged as a powerful anticancer strategy, and small molecules that either inhibit or activate certain proteins in the apoptotic pathway have great potential as novel chemotherapeutic agents. Central to apoptosis is the activation of the zymogen procaspase-3 to caspase-3. Caspase 3 is the key "executioner" caspase, catalyzing the hydrolysis of a multitude of protein substrates within the cell. Interestingly, procaspase-3 levels are often elevated in cancer cells, suggesting a compound that directly stimulates the activation of procaspase-3 to caspase-3 could selectively induce apoptosis in cancer cells. We recently reported the discovery of a compound, PAC-1, which enhances procaspase-3 activity in vitro and induces apoptotic death in cancer cells in culture and in mouse xenograft models. Described herein is the mechanism by which PAC-1 activates procaspase-3 in vitro. We show that zinc inhibits the enzymatic activity of procaspase-3 and that PAC-1 strongly activates procaspase-3 in buffers that contain zinc. PAC-1 and zinc form a tight complex with one another, with a dissociation constant of approximately 42 nM. The combined data indicate that PAC-1 activates procaspase-3 in vitro by sequestering inhibitory zinc ions, thus allowing procaspase-3 to autoactivate itself to caspase-3. The small-molecule-mediated activation of procaspases has great therapeutic potential and thus this discovery of the in vitro mechanism of action of PAC-1 is critical to the development and optimization of other procaspase-activating compounds. PMID- 19281822 TI - A deoxyribozyme, Sero1C, uses light and serotonin to repair diverse pyrimidine dimers in DNA. AB - An in vitro selection search for DNAs capable of catalyzing photochemistry yielded two distinctive deoxyribozymes (DNAzymes) with photolyase activity: UV1C, which repaired thymine dimers within DNA using a UV light of >300 nm wavelength and no extraneous cofactor, and Sero1C, which required the tryptophan metabolite serotonin as cofactor in addition to the UV light. Catalysis by Sero1C conformed to Michaelis-Menten kinetics, and analysis of the action spectrum of Sero1C confirmed that serotonin did indeed serve as a catalytic cofactor rather than as a structural cofactor. Sero1C and UV1C showed strikingly distinct wavelength optima for their respective photoreactivation catalyses. Although the rate enhancements characteristic of the two DNAzymes were similar, the cofactor requiring Sero1C repaired a substantially broader range of substrates compared to UV1C, including thymine, uracil, and a range of chimeric deoxypyrimidine and ribopyrimidine dimers. Similarities and differences in the properties of these two photolyase DNAzymes suggest, first, that the harnessing of less damaging UV light for the repair of photolesions may have been a primordial catalytic activity of nucleic acids, and, second, the broader substrate range of Sero1C may highlight an evolutionary advantage to coopting amino-acid-like cofactors by functionality-poor nucleic acid enzymes. PMID- 19281823 TI - Structure, dynamics and folding of an immunoglobulin domain of the gelation factor (ABP-120) from Dictyostelium discoideum. AB - We have carried out a detailed structural and dynamical characterisation of the isolated fifth repeat of the gelation factor (ABP-120) from Dictyostelium discoideum (ddFLN5) by NMR spectroscopy to provide a basis for studies of co translational folding on the ribosome of this immunoglobulin-like domain. The isolated ddFLN5 can fold autonomously in solution into a structure that resembles very closely the crystal structure of the domain in a construct in which the adjacent sixth repeat (ddFLN6) is covalently linked to its C-terminus in tandem but deviates locally from a second crystal structure in which ddFLN5 is flanked by ddFLN4 and ddFLN6 at both N- and C-termini. Conformational fluctuations were observed via (15)N relaxation methods and are primarily localised in the interstrand loops that encompass the C-terminal hemisphere. These fluctuations are distinct in location from the region where line broadening is observed in ddFLN5 when attached to the ribosome as part of a nascent chain. This observation supports the conclusion that the broadening is associated with interactions with the ribosome surface [Hsu, S. T. D., Fucini, P., Cabrita, L. D., Launay, H., Dobson, C. M. & Christodoulou, J. (2007). Structure and dynamics of a ribosome bound nascent chain by NMR spectroscopy. Proc. Natl. Acad. Sci. USA, 104, 16516 16521]. The unfolding of ddFLN5 induced by high concentrations of urea shows a low population of a folding intermediate, as inferred from an intensity-based analysis, a finding that differs from that of ddFLN5 as a ribosome-bound nascent chain. These results suggest that interesting differences in detail may exist between the structure of the domain in isolation and when linked to the ribosome and between protein folding in vitro and the folding of a nascent chain as it emerges from the ribosome. PMID- 19281824 TI - A relationship between mRNA expression levels and protein solubility in E. coli. AB - Each step in the process of gene expression, from the transcription of DNA into mRNA to the folding and posttranslational modification of proteins, is regulated by complex cellular mechanisms. At the same time, stringent conditions on the physicochemical properties of proteins, and hence on the nature of their amino acids, are imposed by the need to avoid aggregation at the concentrations required for optimal cellular function. A relationship is therefore expected to exist between mRNA expression levels and protein solubility in the cell. By investigating such a relationship, we formulate a method that enables the prediction of the maximal levels of mRNA expression in Escherichia coli with an accuracy of 83% and of the solubility of recombinant human proteins expressed in E. coli with an accuracy of 86%. PMID- 19281825 TI - Kinetic metabolic modelling for the control of plant cells cytoplasmic phosphate. AB - A previously developed kinetic metabolic model for plant metabolism was used in a context of identification and control of intracellular phosphate (Pi) dynamics. Experimental data from batch flask cultures of Eschscholtiza californica cells was used to calibrate the model parameters for the slow dynamics (growth, nutrition, anabolic pathways, etc.). Perturbation experiments were performed using a perfusion small-scale bioreactor monitored by in vivo(31)P NMR. Parameter identification for Pi metabolism was done by measuring the cells dynamic response to different inputs for extracellular Pi (two pulse-response experiments and a step-response experiment). The calibrated model can describe Pi translocation between the cellular pools (vacuole and cytoplasm). The effect of intracellular Pi management on ATP/ADP and phosphomonoesters concentrations is also described by the model. The calibrated model is then used to develop a control strategy on the cytoplasmic Pi pool. From the identification of the systems dynamics, a proportional-integral controller was designed and tuned. The closed-loop control was implemented in the small-scale NMR bioreactor and experimental results were in accordance with model predictions. Thus, the calibrated model is able to predict cellular behaviour for phosphate metabolism and it was demonstrated that it is possible to control the intracellular level of cytoplasmic Pi in plant cells. PMID- 19281826 TI - Oxidative stress and metabolic syndrome. AB - Metabolic syndrome is a collection of cardiometabolic risk factors that includes obesity, insulin resistance, hypertension and dyslipidemia. Although there has been significant debate regarding the criteria and concept of the syndrome, this clustering of risk factors is unequivocally linked to an increased risk of developing type 2 diabetes and cardiovascular disease. Metabolic syndrome is often characterized by oxidative stress, a condition in which an imbalance results between the production and inactivation of reactive oxygen species. Reactive oxygen species can best be described as double-edged swords; while they play an essential role in multiple physiological systems, under conditions of oxidative stress, they contribute to cellular dysfunction. Oxidative stress is thought to play a major role in the pathogenesis of a variety of human diseases, including atherosclerosis, diabetes, hypertension, aging, Alzheimer's disease, kidney disease and cancer. The purpose of this review is to discuss the role of oxidative stress in metabolic syndrome and its major clinical manifestations (namely coronary artery disease, hypertension and diabetes). It will also highlight the effects of lifestyle modification in ameliorating oxidative stress in metabolic syndrome. Discussion will be limited to human data. PMID- 19281827 TI - Use of plasmon waveguide resonance (PWR) spectroscopy for examining binding, signaling and lipid domain partitioning of membrane proteins. AB - AIMS: Due to their anisotropic properties and other factors, it has been difficult to determine the conformational and dynamic properties of integral membrane proteins such as G-protein coupled receptors (GPCRs), growth factor receptors, ion channels, etc. in response to ligands and subsequent signaling. Herein a novel methodology is presented that allows such studies to be performed while maintaining the receptors in a membrane environment. MAIN METHOD: Plasmon waveguide resonance (PWR) spectroscopy is a relatively new biophysical method which allows one to directly observe structural and dynamic changes which occur on interaction of GPCRs (and other integral membrane proteins) with ligands and signaling molecules. The delta opioid receptor (DOR) and its ligands serve as an excellent model system to illustrate the new insights into GPCR signaling that can be obtained by this method. KEY FINDINGS: Among our key findings are: 1) it is possible to obtain the following information directly and without any need for labels (radioactive, fluorescent, etc.): binding affinities, and the ability to distinguish between agonists, antagonists, inverse agonist, and partial agonists without a need for second messenger analysis; 2) it is possible to determine directly, again without a need for labels, G-protein binding to variously occupied or unoccupied DORs, and to determine which alpha-subtype is involved in allowing structurally different agonist ligands to have differential effects; 3) GTPgammaS binding can be examined directly; and 4) binding of the DOR with different ligands leads to differential segregation of the ligand-receptor complex into lipid rafts. SIGNIFICANCE: The implications of these discoveries suggest a need to modify our current views of GPCR-ligand interactions and signaling. PMID- 19281828 TI - On the interaction of fluorophore-encapsulating PEGylated lecithin liposomes with hamster and human platelets. AB - Polyethylene glycol (PEG)-grafted phosphatidylcholine liposomes are used as drug carriers due to their low immunogenicity and prolonged circulation time. The interaction between sterically stabilized lecithin liposomes and platelets has not been investigated before, and deserves to be subjected to scrutiny inasmuch as the uptake of liposomes by platelets could be detrimental for drug delivery and primary hemostasis. Consequently, the interaction between resting and convulxin-activated hamster and human platelets and calcein- or 5,6 carboxyfluorescein-encapsulating PEGylated liposomes composed of distearoyl- and dipalmitoyl phosphatidylcholine and PEG-derivatized distearoyl phosphatidylethanolamine was investigated by flow cytometry, confocal microscopy, and a glass capillary thrombosis model. Fluorescently labeled liposomes of the same composition were subsequently assayed in vivo after 15 and 45 min of systemic circulation. Neither resting nor activated hamster and human platelets interacted with liposomes at 0.70 mM lipid concentration. An absence of any interaction was corroborated in the in vivo experiments. Alternatively, flow cytometry assays evinced that human platelets interact with liposomes at lipid concentrations of >or=1.35 mM. These interactions were more profound for activated platelets than resting platelets. We conclude that the use of PEGylated lecithin liposomes at lipid concentrations of <1.35 mM has no detrimental impact on liposomal drug delivery based on PEGylated lecithin liposomes, but that these drug carriers may be associated with a reduced targeting efficacy or compromised primary hemostatic system when used at concentrations of >or=1.35 mM. In contrast, these drug carriers may become valuable in thrombosis- and drug delivery-related research and applications at concentrations of >or=1.35 mM. PMID- 19281829 TI - Photoreceptor degeneration and retinal inflammation induced by very low-density lipoprotein receptor deficiency. AB - Our previous studies have shown that very low-density lipoprotein receptor (VLDLR) is a negative regulator of the Wnt pathway. The present study showed that VLDLR gene knockout (Vldlr(-/-)) mice displayed impaired cone ERG responses at early ages. Immunostaining of mid-wavelength cones showed significantly decreased cone densities in the retina and shortened cone outer segments in Vldlr(-/-) mice. At older ages, Vldlr(-/-) mice displayed declined rod ERG responses, decreased layers of photoreceptor nuclei, reduced rhodopsin levels and decreased levels of 11-cis retinal, the chromophore of visual pigments. As shown by fluorescein angiography and permeability assay, Vldlr(-/-) mice had severe retinal vascular leakage. ZO-1, a tight junction protein, was down-regulated in Vldlr(-/-) mouse retinae, further supporting the impaired blood-retinal barrier. Double staining of pericytes and endothelial cells in retinal sections revealed that neovasculature in Vldlr(-/-) mice lacks pericyte coverage, suggesting impaired maturation of retinal vasculature in Vldlr(-/-) mice. Staining of adherent leukocytes in the retinal vasculature revealed significant leukostasis in Vldlr(-/-) mice. Moreover, Vldlr(-/-) mice displayed up-regulated expression of multiple pro-inflammatory factors and activated NF-kappaB and HIF-1 alpha, key regulators of inflammation. These findings suggest that deficiency of VLDLR leads to retinal degeneration and inflammation. PMID- 19281830 TI - IL-20 activates human lymphatic endothelial cells causing cell signalling and tube formation. AB - IL-20 is an arteriogenic cytokine that remodels collateral networks in vivo, and plays a role in cellular organization. Here, we investigate its role in lymphangiogenesis using a lymphatic endothelial cell line, hTERT-HDLEC, which expresses the lymphatic markers LYVE-1 and podoplanin. Upon stimulation of hTERT HDLEC with IL-20, we found an increase in the intracellular free calcium concentration, in Akt and eNOS phosphorylations as well as in perinuclear NO production. We found that eNOS phosphorylation and NO synthesis are highly dependent on the PI3K/Akt signalling pathway. We also found an IL-20 induced phosphorylation of Erk1/2 and mTOR, and using the MEK inhibitor PD98059 and mTOR complex inhibitor rapamycin we demonstrated the importance of these signalling pathways in IL-20-mediated proliferation. IL-20 triggered actin polymerization and morphological changes resulting in elongated cell structures, and in matrigels, IL-20 caused tube formations of hTERT-HDLEC in a PI3K- and mTOR dependent way. In a sprouting assay we found that IL-20 caused cell migration within 24 h at a rate comparable to VEGF-C, and this migration could be inhibited by wortmannin and rapamycin. These data show that IL-20 activates cell signalling resulting in lymphangiogenic processes including migration, proliferation and tube formation. Thus, IL-20 is a cytokine that has the potential of activating or modulating the formation of lymphatic vessels. PMID- 19281831 TI - Behavioral effects of ventilated micro-environment housing in three inbred mouse strains. AB - Animal facilities aim to combine animal welfare with cost-efficiency and limited care staff requirements, and individually ventilated cage (IVC) systems were developed towards these goals. While IVC have great sanitary advantages both for the animals but also for the care staff, these systems involve potentially deleterious features such as high levels of air renewal, noise, and subtle vibrations of the racks because of the air filtering system used, but also reduce the frequency of stressful cage changes. It is unknown in how far these conditions may influence the animals' behavior. This issue becomes critical as many facilities are switching to IVC systems, possibly complicating replication of data or biasing ongoing studies. We investigated the effects of IVC housing in mice on different behaviors including anxiety, exploration, and learning in males and females of three common and phenotypically distant strains. Results demonstrate robust effects of IVC in multiple behavioral tests with the direction of the effect strongly dependent on strain and sex. These data should serve to alert researchers that a switch to IVC housing during the course of an experiment has the potential to bias results in a serious manner. In addition, behavioral baseline data will have to be re-established once the switch has been completed. PMID- 19281832 TI - Activation of ROS/NF-kappaB and Ca2+/CaM kinase II are necessary for VCAM-1 induction in IL-1beta-treated human tracheal smooth muscle cells. AB - Histone acetylation regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs) plays a critical role in the expression of inflammatory genes, such as vascular cell adhesion molecule-1 (VCAM-1). Oxidative processes have been shown to induce VCAM-1 expression. Here, we investigated the mechanisms underlying IL-1beta-induced VCAM-1 expression in human tracheal smooth muscle cells (HTSMCs). Our results showed that IL-1beta enhanced HTSMCs-monocyte adhesion through up-regulation of VCAM-1, which was inhibited by pretreatment with selective inhibitors of PKCalpha (Go6976), c-Src (PP1), NADPH oxidase [diphenylene iodonium (DPI) and apocynin (APO)], intracellular calcium chelator (BAPTA/AM), PI-PLC (U73122), CaM (calmidazolium chloride), CaM kinase II (KN62), p300 (garcinol), NF-kappaB (Bay11-7082), HDAC (trichostatin A), and ROS scavenger [N-acetyl-L-cysteine (NAC)] or transfection with siRNAs of MyD88, PKCalpha, Src, p47(phox), p300, and HDAC4. Moreover, IL-1beta stimulated NF-kappaB and CaMKII phosphorylation through MyD88-dependent PI-PLC/PKCalpha/c-Src/ROS and PI PLC/Ca2+/CaM pathways, respectively. Activation of NF-kappaB and CaMKII may eventually lead to the acetylation of histone residues and phosphorylation of histone deacetylases. These findings suggested that IL-1beta induced VCAM-1 expression via these multiple signaling pathways in HTSMCs. Blockade of these pathways may reduce monocyte adhesion via VCAM-1 suppression and attenuation of the inflammatory responses in airway diseases. PMID- 19281833 TI - Embryotoxic potential of N-methyl-pyrrolidone (NMP) and three of its metabolites using the rat whole embryo culture system. AB - N-methyl-2-pyrrolidone (NMP), which undergoes extensive biotransformation, has been shown in vivo to cause developmental toxicity and, especially after oral treatment, malformations in rats and rabbits. Data are lacking as to whether the original compound or one of its main metabolites is responsible for the toxic effects observed. Therefore, the relative embryotoxicity of the parent compound and its metabolites was evaluated using rat whole embryo culture (WEC) and the balb/c 3T3 cytotoxicity test. The resulting data were evaluated using two strategies; namely, one based on using all endpoints determined in the WEC and the other including endpoints from both the WEC and the cytotoxicity test. On basis of the first analysis, the substance with the highest embryotoxic potential is NMP, followed by 5-hydroxy-N-methyl-pyrrolidone (5-HNMP), 2-hydroxy-N methylsuccinimide (2-HMSI) and N-methylsuccinimide (MSI). Specific dysmorphogeneses induced by NMP and 5-HNMP were aberrations in the head region of the embryos, abnormal development of the second visceral arches and open neural pores. The second evaluation strategy used only two endpoints of the WEC, i.e. the no observed adverse effect concentration (NOAEC(WEC)) and the lowest concentration leading to dysmorphogenesis in 100% of the cultured embryos (IC(Max WEC)). In addition to these WEC endpoints the IC(50 3T3) from the cytotoxicity test (balb/c 3T3 fibroblasts) was included in the evaluation scheme. These three endpoints were applied to a prediction model developed during a validation study of the European Centre for the Validation of Alternative Methods (ECVAM) allowing the classification of the embryotoxic potential of each compound into three classes (non-, weakly- and strongly embryotoxic). Consistent results from both evaluation strategies were observed, whereby NMP and its metabolites revealed a direct embryotoxic potential. Hereby, only NMP and 5-HNMP induced specific embryotoxic effects and were classified as weakly embryotoxic, whereas the other two metabolites, 2-HMSI and MSI, were determined to be non-embryotoxic. PMID- 19281834 TI - In vivo effects of cnidarian toxins and venoms. AB - Cnidarians (Coelenterates), a very old and diverse animal phylum, possess a wide variety of biologically active substances that can be considered as toxins. Anthozoan toxins can be classified into two chemically very different groups, namely polypeptide toxins isolated from sea anemones and diterpenes isolated from octocorals. Cubozoan and scyphozoan protein toxins have been the most elusive cnidarian toxins to investigate - despite a tremendous effort in the past few decades, very few of these large, relatively unstable protein toxins were isolated, but recently this has been achieved for cubozoan venoms. Hydrozoans mainly contain large proteins with physiological mechanisms of action similar to the sea anemone and jellyfish pore-forming toxins. This article will focus on the in vivo physiological effects of cnidarian toxins and venoms; their actions at the cellular level will only be considered to understand their actions at the organ and whole animal levels. An understanding of mechanisms underlying the in vivo toxic effects will facilitate the development of more effective treatments of cnidarian envenomations. PMID- 19281836 TI - Simulation analysis of bandpass filtering properties of a rod photoreceptor network. AB - The bandpass filtering properties of a rod network were studied via computer simulations. Sinusoidal current stimuli were applied to a single rod model to characterize its temporal filtering properties. The simulated frequency response revealed that a single rod behaves as a bandpass filter whose characteristics are affected by the stimulus strength and frequency. We analyzed the contribution of individual ionic currents to bandpass filtering and found that the filtering of small signals is largely regulated by the calcium-dependent currents I(K(Ca)) and I(Cl(Ca)), whereas the filtering of large signals is regulated by the hyperpolarization-activated current, I(h). Furthermore, rod network modeling by electrically interconnecting the single rod models revealed that the acceleration of signals that spread laterally through the rod network is attributed to I(K(Ca)) and not I(h). PMID- 19281835 TI - Actions of octocoral and tobacco cembranoids on nicotinic receptors. AB - Nicotinic acetylcholine receptors (AChRs) are pentameric proteins that form agonist-gated cation channels through the plasma membrane. AChR agonists and antagonists are potential candidates for the treatment of neurodegenerative diseases. Cembranoids are naturally occurring diterpenoids that contain a 14 carbon ring. These diterpenoids interact with AChRs in complex ways: as irreversible inhibitors at the agonist sites, as noncompetitive inhibitors, or as positive modulators, but no cembranoid was ever shown to have agonistic activity on AChRs. The cembranoid eupalmerin acetate displays positive modulation of agonist-induced currents in the muscle-type AChR and in the related gamma aminobutyric acid (GABA) type A receptor. Moreover, cembranoids display important biological effects, many of them mediated by nicotinic receptors. Cembranoids from tobacco are neuroprotective through a nicotinic anti-apoptotic mechanism preventing excitotoxic neuronal death which in part could result from anti inflammatory properties of cembranoids. Moreover, tobacco cembranoids also have anti-inflammatory properties which could enhance their neuroprotective properties. Cembranoids from tobacco affect nicotine-related behavior: they increase the transient initial ataxia caused by first nicotine injection into naive rats and inhibit the expression of locomotor sensitization to repeated injections of nicotine. In addition, cembranoids are known to act as anti-tumor compounds. In conclusion, cembranoids provide a promising source of lead drugs for many clinical areas, including neuroprotection, smoking-cessation, and anti cancer therapies. PMID- 19281837 TI - Stimulant-induced changes in smoking and caloric intake: influence of rate of onset. AB - Rate-of-onset modulates the subject-rated effects of stimulants. Results of two studies from our laboratory demonstrate that immediate-release methylphenidate increases smoking and decreases caloric intake. Whether rate-of-onset influences the effects of methylphenidate on smoking and eating is unknown. The present experiment examined the influence of a range of doses of immediate- (7.5-30 mg) and sustained-release (18-72 mg) methylphenidate as well as placebo on smoking and eating. Eight cigarette smokers participated. A double-dummy drug administration procedure was used to maintain the double blind because immediate release methylphenidate produces peak plasma concentrations 1.5-2 h and the sustained-release formulation produces peak plasma concentrations 6-8 h after oral administration. Smoking and eating were assessed for 4 h across the predicted peak effects of both methylphenidate formulations. Measures of smoking included total cigarettes, puffs, and carbon monoxide levels. Snacks and decaffeinated beverages were available ad libitum and caloric intake was monitored during the four-hour smoking session. Immediate- and sustained-release methylphenidate increased smoking and decreased caloric intake. The effects of methylphenidate generally did not vary as a function of formulation. The results of this study may have important implications for the treatment of disorders that require stimulant medications. Smoking should be monitored in patients that are prescribed stimulant medications, regardless of the formulation type. PMID- 19281838 TI - Ethanol-mediated aversive learning as a function of locomotor activity in a novel environment in infant Sprague-Dawley rats. AB - Unlike adult heterogeneous rats, infant rats are sensitive to ethanol's locomotor stimulating effects. Susceptibility to this ethanol effect varies as a function of baseline locomotor activity levels. Infant rats with higher baseline activity levels are more sensitive to ethanol's stimulating effects than those with lower baseline activity levels. The present study was designed to analyze susceptibility to ethanol-induced motivational learning in subpopulations of infant heterogeneous rats that differ in baseline activity in a novel environment. On postnatal day 11 (PD 11) baseline locomotor activity was registered and infants were divided into high and low responders (HR, LR). In Experiment 1, pups were trained in a procedure of conditioned taste aversion employing ethanol (0.0, 0.5 or 2.5 g/kg) as unconditioned stimulus (US) and saccharin as conditioned stimulus. In Experiment 2 the same procedure was employed with LiCl (0.0, 0.25 or 0.5% of body weight of a 0.3 M LiCl solution) as US. HR were more resistant to the aversive effects of ethanol than LR while magnitude of LiCl-induced conditioned taste aversion was similar in HR and LR. These results suggest the possibility of early detection of subpopulations of rats with differential sensitivity to ethanol's effects. PMID- 19281839 TI - Effect of lidocaine administration at the nucleus locus coeruleus level on lateral hypothalamus-induced antinociception in the rat. AB - Several lines of evidence have shown that stimulation or inactivation of lateral hypothalamus (LH) produces antinociception. In this study, we assessed the role of nucleus locus coeruleus (LC) in antinociceptive response induced by LH stimulation or inactivation in the rat. The cholinergic agonist carbachol (125 nmol/0.5 microl saline) or lidocaine (2%; 0.5 microl) was unilaterally microinjected into the LH with the LC inactivation concurrently. Antinociceptive responses were obtained by tail-flick test and represented as maximal possible effect (MPE) at 5, 10, 15, 20, 30 and 60 min after drug administration. The results showed that microinjection of carbachol into the LH significantly induced antinociception at 5 and 10 min (p<0.001). This effect was significantly blocked by microinjection of lidocaine into the LC. On the other hand, microinjection of lidocaine into LH-induced antinociception at 5 (p<0.01) and 10 (p<0.05) min after administration. However, inactivation of the LC following the LH inactivation increased MPE at 5 min after injection. These findings support the conclusion that antinociception produced by LH stimulation or inactivation involves two separate mechanisms. It seems that analgesic response induced by LH stimulation is mediated in part by the subsequent activation of spinally projecting noradrenergic neurons in the LC cell group. PMID- 19281840 TI - A strategy to verify the absence of the pgm locus in Yersinia pestis strain candidates for select agent exemption. AB - Yersinia pestis is a Department of Health and Human Services select agent as defined in federal regulations. Certain attenuated strains of Y. pestis, such as the pgm(-) strain, are exempt from these regulations. Herein we describe a strategy to verify the absence of the pgm locus in Y. pestis strains being considered as candidates for select agent exemption by PCR analysis of virulence associated genes. PMID- 19281841 TI - Identification of distinct ciprofloxacin susceptibility in Acinetobacter spp. by detection of the gyrA gene mutation using real-time PCR. AB - Ciprofloxacin-resistant (CipR) Acinetobacter spp. was associated with a mutation in quinolone resistance-determining region (QRDR) of gyrA gene from Ser83 to Leu83. A total of 54 Acinetobacter baumannii, 11 A. genospecies 3, and 17 A. genospecies 13TU clinical isolates were determined for ciprofloxacin susceptibility and their QRDR sequenced. Most of A. baumannii were CipR and had a mutated QRDR of gyrA gene, and all A. genospecies 3 and 13TU isolates were ciprofloxacin-susceptible (CipS) and had a wild-type QRDR of gyrA gene. A real time PCR assay was developed to rapidly differentiate the CipR and CipS Acinetobacter spp. This assay was based on probe hybridization followed by melting temperature (T(m)) analysis to discriminate the QRDR of gyrA gene. All CipR strains had a T(m) at 47 degrees C, and most of CipS strains had a higher T(m) at 51.5 degrees C. Four CipS A. genospecies 3 isolates with an A base at 264nt of the gyrA gene had a T(m) at 49.5 degrees C. The assay can rapidly and accurately identify the mutated QRDR of gyrA gene in CipR Acinetobacter spp., and potentially increase the rate of the appropriate therapy for Acinetobacter infections. PMID- 19281842 TI - Hydrogen peroxide impairs insulin-stimulated assembly of mTORC1. AB - Oxidants are well recognized for their capacity to reduce the phosphorylation of the mammalian target of rapamycin (mTOR) substrates, eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and p70 S6 kinase 1 (S6K1), thereby hindering mRNA translation at the level of initiation. mTOR functions to regulate mRNA translation by forming the signaling complex mTORC1 (mTOR, raptor, GbetaL). Insulin signaling to mTORC1 is dependent upon phosphorylation of Akt/PKB and the inhibition of the tuberous sclerosis complex (TSC1/2), thereby enhancing the phosphorylation of 4E-BP1 and S6K1. In this study we report the effect of H(2)O(2) on insulin-stimulated mTORC1 activity and assembly using A549 and bovine aortic smooth muscle cells. We show that insulin stimulated the phosphorylation of TSC2 leading to a reduction in raptor-mTOR binding and in the quantity of proline-rich Akt substrate 40 (PRAS40) precipitating with mTOR. Insulin also increased 4E-BP1 coprecipitating with mTOR and the phosphorylation of the mTORC1 substrates 4E-BP1 and S6K1. H(2)O(2), on the other hand, opposed the effects of insulin by increasing raptor-mTOR binding and the ratio of PRAS40/raptor derived from the mTOR immunoprecipitates in both cell types. These effects occurred in conjunction with a reduction in 4E-BP1 phosphorylation and the 4E-BP1/raptor ratio. siRNA-mediated knockdown of PRAS40 in A549 cells partially reversed the effect of H(2)O(2) on 4E-BP1 phosphorylation but not on S6K1. These findings are consistent with PRAS40 functioning as a negative regulator of insulin-stimulated mTORC1 activity during oxidant stress. PMID- 19281843 TI - 6-mercaptopurine (6-MP) induces p53-mediated apoptosis of neural progenitor cells in the developing fetal rodent brain. AB - 6-mercaptopurine (6-MP), a DNA-damaging agent, induces apoptosis of neural progenitor cells, and causes malformation in the fetal brain. The aim of the present study is to clarify the molecular pathway of 6-MP-induced apoptosis of neural progenitor cells in the fetal telencephalon of rats and mice. p53 protein is activated by DNA damage and induces apoptosis through either the intrinsic pathway involving the mitochondria or the extrinsic pathway triggered by death receptors. In this study, the expression of puma and cleaved caspase-9 proteins, which are specific intrinsic pathway factors, increased in the rat telencephalon after 6-MP treatment. 6-MP-induced apoptosis of neural progenitor cells was completely absent in p53-deficient mice. On the other hand, the expression of Fas protein, an extrinsic pathway factor, did not change throughout the experimental period in the rat telencephalon treated with 6-MP. The number of apoptotic neural progenitor cells was similar among Fas-mutated lpr/lpr and wild-type mice, suggesting that the Fas pathway does not play a significant role in 6-MP-induced apoptosis of neural progenitor cells. These results may suggest that the p53 mediated intrinsic pathway is essential for 6-MP-induced apoptosis of neural progenitor cells in the developing telencephalon of rats and mice. PMID- 19281844 TI - Cytoplasmic prion protein induces forebrain neurotoxicity. AB - The prion protein (PrP) is essential for the pathogenesis of prion disease. PrP has been detected in the cytosol of neurons and transgenic mice expressing PrP in the cytosol (cyPrP) under a pan-neuronal promoter developed rapid cerebellar granule neuron degeneration. Yet, it remains unclear whether cyPrP is capable to cause toxicity in other neuronal populations. Here, we report that transgenic mice expressing cyPrP in the forebrain neurons developed behavioral abnormalities including clasping and hyperactivity. These mice had reduced thickness in cortex and developed astrogliosis in hippocampal and cortical regions. Moreover, cyPrP in these mice was recognized by the A11 anti-oligomer antibody and was associated with the hydrophobic lipid core of membranes, indicating that cyPrP oligomer caused membrane perturbation contributes to cyPrP neurotoxicity. Together, our results clearly revealed that cyPrP is able to cause toxicity in different neuronal populations, supporting a role of cyPrP in PrP-mediated neurodegenerative disorders. PMID- 19281845 TI - The use of Eudragit RS 100/cyclodextrin nanoparticles for the transmucosal administration of glutathione. AB - The aim of this work was to develop and characterize new nanoparticle systems based on Eudragit RS 100 and cyclodextrins (CDs) for the transmucosal administration of glutathione (GSH). For this purpose, nanoparticles (NPs) with the mucoadhesive properties of Eudragit RS 100 and the penetration enhancing and peptide protective properties of CDs were prepared and evaluated. The quasi emulsion solvent diffusion technique was used to prepare the NPs with natural and chemically modified (HP-beta-CD and Me-beta-CD) CDs. The NPs prepared showed homogeneous size distribution, mean diameters between 99 and 156nm, a positive net charge and spherical morphology. Solid state FT-IR, thermal analysis (DSC), and X-ray diffraction studies suggest that the nanoencapsulation process produces a marked decrease in crystallinity of GSH. The encapsulation efficiency of the peptide was found to be between 14.8% and 24%. The results indicate that mean diameters, surface charges and drug-loaded NPs were not markedly affected by the CD, whereas the presence of the latter influences drug release and to some extent peptide stability and absorption. Finally, it has been shown that CD/Eudragit RS 100 NPs may be used for transmucosal absorption of GSH without any cytotoxicity using the epithelial human HaCaT and murine monocyte macrophage RAW264.7 cell lines. PMID- 19281846 TI - Phosphodiesterase 10 inhibition reduces striatal excitotoxicity in the quinolinic acid model of Huntington's disease. AB - Decreased activity of cAMP responsive element-binding protein (CREB) is thought to contribute to the death of striatal medium spiny neurons in Huntington's disease (HD). Therefore, therapies that increase levels of activated CREB, may be effective in fighting neurodegeneration in HD. In this study, we sought to determine whether the phosphodiesterase type 10 (PDE10A) inhibitor TP10 exerts a neuroprotective effect in an excitotoxic model of HD. Rats were surgically administered with quinolinic acid into striatum and subsequently treated with TP10 daily for two or eight weeks. After 2 weeks of TP10 treatment, striatal lesion size was 52% smaller and the surviving cell number was several times higher than in the vehicle-treated group. These beneficial effects of TP10 were maintained through 8 weeks. TP10 treatment also increased significantly the levels of activated CREB in the striatal spiny neurons, which is hypothesized to be a contributing mechanism for the neuroprotective effect. Our findings suggest PDE10A inhibition as a novel neuroprotective approach to the treatment of HD and confirm the importance of phosphodiesterase inhibition in fighting the disease. PMID- 19281847 TI - Mutations in amyloid precursor protein affect its interactions with presenilin/gamma-secretase. AB - Alzheimer's disease is characterized by accumulation of toxic beta-amyloid (Abeta) in the brain and neuronal death. Several mutations in presenilin (PS1) and beta-amyloid precursor protein (APP) associate with an increased Abeta(42/40) ratio. Abeta(42), a highly fibrillogenic species, is believed to drive Abeta aggregation. Factors shifting gamma-secretase cleavage of APP to produce Abeta(42) are unclear. We investigate the molecular mechanism underlying altered Abeta(42/40) ratios associated with APP mutations at codon 716 and 717. Using FRET-based fluorescence lifetime imaging to monitor APP-PS1 interactions, we show that I716F and V717I APP mutations increase the proportion of interacting molecules earlier in the secretory pathway, resulting in an increase in Abeta generation. A PS1 conformation assay reveals that, in the presence of mutant APP, PS1 adopts a conformation reminiscent of FAD-associated PS1 mutations, thus influencing APP binding to PS1/gamma-secretase. Mutant APP affects both intracellular location and efficiency of APP-PS1 interactions, thereby changing the Abeta(42/40) ratio. PMID- 19281848 TI - E. coli expression of a soluble, active single-chain antibody variable fragment containing a nuclear localization signal. AB - Single-chain antibody variable fragment (scFv) proteins consist of an antibody heavy chain variable sequence joined via a flexible linker to a light chain variable sequence. Prior work has shown that ScFv 18-2 binds the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and sensitizes cancer cells to radiation following nuclear microinjection. A potential clinical delivery strategy is based on modification of the scFv so that it can be taken up into cells and imported to the nucleus. This will require development of an expression system for a nuclear localization signal (NLS)-tagged scFv derivative. We found, however, that addition of the highly basic NLS severely compromised expression in the host-vector system used for the parental scFv. After testing a variety of host strains, fusion partners, and NLS sequences and placements, successful expression was obtained with a construct containing a stabilizing N-terminal maltose binding protein tag and a single, optimized, C-terminal NLS moiety. Amylose affinity-purified ScFv 18-2 NLS protein was stable to storage at 4 degrees C in the presence of glycerol or trehalose, bound selectively to an epitope peptide, and was cleavable at an engineered Factor Xa protease site. Following lipid-mediated uptake into cultured cells, NLS-tagged ScFv 18-2, unlike the parental ScFv 18-2, localized predominantly in the cell nucleus. PMID- 19281849 TI - Dissecting medial temporal lobe contributions to item and associative memory formation. AB - A fundamental and intensively discussed question is whether medial temporal lobe (MTL) processes that lead to non-associative item memories differ in their anatomical substrate from processes underlying associative memory formation. Using event-related functional magnetic resonance imaging, we implemented a novel design to dissociate brain activity related to item and associative memory formation not only by subsequent memory performance and anatomy but also in time, because the two constituents of each pair to be memorized were presented sequentially with an intra-pair delay of several seconds. Furthermore, the design enabled us to reduce potential differences in memory strength between item and associative memory by increasing task difficulty in the item recognition memory test. Confidence ratings for correct item recognition for both constituents did not differ between trials in which only item memory was correct and trials in which item and associative memory were correct. Specific subsequent memory analyses for item and associative memory formation revealed brain activity that appears selectively related to item memory formation in the posterior inferior temporal, posterior parahippocampal, and perirhinal cortices. In contrast, hippocampal and inferior prefrontal activity predicted successful retrieval of newly formed inter-item associations. Our findings therefore suggest that different MTL subregions indeed play distinct roles in the formation of item memory and inter-item associative memory as expected by several dual process models of the MTL memory system. PMID- 19281850 TI - 3 T MRI relaxometry detects T2 prolongation in the cerebral normal-appearing white matter in multiple sclerosis. AB - MRI at 3 T has increased sensitivity in detecting overt multiple sclerosis (MS) brain lesions; a growing body of data suggests clinically relevant damage occurs in the normal-appearing white matter (NAWM). We tested a novel pulse sequence to determine whether 3 T MRI spin-spin relaxometry detected damage in NAWM of MS patients (n=13) vs. age-matched normal controls [(NL) (n=11)]. Baseline characteristics of the MS group were: age (mean+/-SD) 42.5+/-5.4 (range 33-51 years), disease duration 9.0+/-6.4 (range 1-22 years), Expanded Disability Status Scale score 2.5+/-1.7 (range 1-6.5). Brain MRI measures, obtained at 3 T, included global and regional NAWM transverse relaxation rate [R2 (=1/T2)], derived from 3D fast spin-echo T2 prepared images, and global white matter volume fraction derived from SPGR images. The regional NAWM areas investigated were the frontal lobe, parietal lobe, and the genu and splenium of the corpus callosum. Mean NAWM R2 was lower (indicating T2 prolongation) in MS than NL in the whole brain (p=0.00047), frontal NAWM (p=0.00015), parietal NAWM (p=0.0069) and callosal genu (p=0.0019). Similarly, R2 histogram peak position was lower in NAWM in MS than NL in the whole brain (p=0.019). However, the normalized WM volume fractions were similar in both MS and NL (p>0.1). This pilot study suggests that a novel 3D fast spin-echo pulse sequence at 3 T, used to derive R2 relaxation maps, can detect tissue damage in the global and regional cerebral NAWM of MS patients that is missed by conventional lesion and atrophy measures. Such findings may represent demyelination, inflammation, glial proliferation and axonal loss. PMID- 19281851 TI - The impact of aging and gender on brain viscoelasticity. AB - Viscoelasticity is a sensitive measure of the microstructural constitution of soft biological tissue and is increasingly used as a diagnostic marker, e.g. in staging liver fibrosis or characterizing breast tumors. In this study, multifrequency magnetic resonance elastography was used to investigate the in vivo viscoelasticity of healthy human brain in 55 volunteers (23 females) ranging in age from 18 to 88 years. The application of four vibration frequencies in an acoustic range from 25 to 62.5 Hz revealed for the first time how physiological aging changes the global viscosity and elasticity of the brain. Using the rheological springpot model, viscosity and elasticity are combined in a parameter mu that describes the solid-fluid behavior of the tissue and a parameter alpha related to the tissue's microstructure. It is shown that the healthy adult brain undergoes steady parenchymal 'liquefaction' characterized by a continuous decline in mu of 0.8% per year (P<0.001), whereas alpha remains unchanged. Furthermore, significant sex differences were found with female brains being on average 9% more solid-like than their male counterparts rendering women more than a decade 'younger' than men with respect to brain mechanics (P=0.016). These results set the background for using cerebral multifrequency elastography in diagnosing subtle neurodegenerative processes not detectable by other diagnostic methods. PMID- 19281852 TI - Semantic information alters neural activation during transverse patterning performance. AB - Memory tasks can be performed using multiple cognitive strategies, which are mediated by different brain systems. The transverse patterning (TP) task is dependent upon the integrity of the hippocampal system, however, we previously demonstrated successful TP following hippocampal damage using meaningful stimuli and relations (Moses, S.N., Ostreicher, M.L., Rosenbaum, R.S., Ryan, J.D., 2008. Successful transverse patterning in amnesia using semantic knowledge. Hippocampus 18, 121-124). Here, we used magnetoencephalgraphy (MEG) to directly observe the neural underpinnings of TP, and the changes that occur as stimuli and relations become more meaningful. In order to optimize our ability to detect signal from deep, non-dominant, brain sources we implemented the event-related synthetic aperture magnetometry minimum-variance beamformer algorithm (ER-SAM; Cheyne, D., Bakhtazad, L., Gaetz, W., 2006. Spatiotemporal mapping of cortical activity accompanying voluntary movements using an event-related beamforming approach. Human Brain Mapping 27, 213-229) coupled with the partial least squares (PLS) multivariate statistical approach (McIntosh, A.R., Bookstein, F.L., Haxby, J.V., Grady, C.L., 1996. Spatial pattern analysis of function brain images using partial least squares. NeuroImage 3, 143-157; McIntosh, A.R., Lobaugh, N.J., 2004. Partial least squares analysis of neuroimaging data: Applications and advances. NeuroImage 23, S250-S263). We found that increased meaningfulness elicited reduced bilateral hippocampal activation, along with increased activation of left prefrontal and temporal cortical structures, including inferior frontal (IFG), as well as anterior temporal and perirhinal cortices. These activation patterns may represent a shift towards reliance upon existing semantic knowledge. This shift likely permits successful TP performance with meaningful stimuli and relations following hippocampal damage. PMID- 19281853 TI - Effects of methylphenidate hydrochloride on the cardiovascular system in vivo and in vitro: a safety pharmacology study. AB - INTRODUCTION: : We examined the effects of methylphenidate hydrochloride (MPH) on the cardiovascular system using in vivo and in vitro study methods in accordance with the ICH-S7B guideline. METHODS: MPH was orally administered at doses of 3, 10 and 30 mg/kg to unrestrained conscious dogs implanted with a telemetry transmitter and attached with body surface electrodes, and electrocardiogram (ECG) leads. The QTcF interval was determined while heart rate (HR), and blood pressure (BP) were measured. Action potentials in isolated guinea-pig papillary muscle and the rapid component of the delayed rectifier potassium current (I(Kr)) in HEK-293 cells stably transfected with hERG were also investigated at concentrations of 0.1, 0.3 and 1 microg/mL (0.37, 1.1 and 3.7 micromol/L) of MPH. RESULTS: No ECG changes were observed except for a shortening of the QT interval due to a shortening of the RR interval at the maximum dose tested, 30 mg/kg. The only observed change was an elevation of BP in dogs at the dose of 30 mg/kg, which is approximately 10 times higher than the maximum therapeutic dose for use in children with attention deficit hyperactivity disorder (ADHD). Neither APD prolongation nor I(Kr) inhibition was observed by MPH in the in vitro studies up to the maximum concentration tested, 1 microg/mL (3.7 micromol/L), which is approximately 34 times higher than the clinically attainable unbound plasma MPH concentrations in children with ADHD. DISCUSSION: These results suggest that it is unlikely that MPH affects ventricular repolarization processes at the therapeutically recommended dose levels in patients with ADHD. PMID- 19281854 TI - Relative expression of cell growth regulatory genes insulin-like growth factors (IGF-1 and IGF-2) and their receptors (IGF-1R and IGF-2R) in somatic cell nuclear transferred (SCNT) and in vitro fertilized (IVF) pre-implantation buffalo embryos. AB - Relative mRNA transcript expression of insulin-like growth factors, IGF-1, IGF-2 and their receptors, IGF-1R and IGF-2R, was studied in SCNT and IVF buffalo embryos at different developmental stages using SYBR green with real-time PCR. SCNT embryos were produced by enucleating IVM oocytes and transferring granulosa cells (passage 5) followed by the electrofusion and chemical activation method. IVF embryos were produced by culturing 15-20 COCs in BO capacitated sperms from frozen and thawed buffalo semen. SCNT embryo production rate was lower than IVF. IGF-1 mRNA expression was significantly upregulated at 2-cell, 16-cell, morula and blastocyst stages of SCNT embryos than in IVF embryos. IGF-1R expression declined from the 2-cell to the 16-cell stage, which increased in later stages and was highest in IVF blastocysts. Similar regulation was observed at different stages in SCNT embryos, except at the 4-cell stage where expression was higher. IGF-2 expression decreased up to the 8-cell stage and increased until the blastocyst stage, being higher in SCNT than IVF embryos. IGF-2R mRNA transcript expression was consistently lower in SCNT than in IVF embryos. Reprogramming of IGF-1, IGF-1R, IGF-2 and IGF-2R expression may have a significant role in cell proliferation in cloned embryos and is developmentally regulated. PMID- 19281855 TI - The polarisome component SPA-2 localizes at the apex of Neurospora crassa and partially colocalizes with the Spitzenkorper. AB - In fungal hyphae multiple protein complexes assemble at sites of apical growth to maintain cell polarity and promote nucleation of actin. Polarity allows the directional traffic of vesicles to the Spitzenkorper (Spk) prior to fusing with the plasma membrane to provide precursors and enzymes required for cell extension and nutrition. One of these complexes is the polarisome, which in Saccharomyces cerevisiae contains Spa2p, Pea2p, Bud6p/Aip3p and Bni1p. To investigate the localization and role of the polarisome during Spk establishment in Neurospora crassa we tagged SPA-2 with the green fluorescent protein (GFP) and examined growing cells by laser scanning confocal microscopy in elongating germ tubes and mature hyphae. SPA-2-GFP accumulated gradually at the apex of germ tubes, when a FM4-64 stained Spk was not still detectable. When the germlings reached about 40microm in length, a FM4-64 stained Spk started to be apparent and from this point on SPA-2-GFP was observed in the apical region of both germ tubes and mature hyphae, as a hand fan shape with a brighter spot at the base. Fusion of the N. crassa SPA-2-GFP strain with a N. crassa strain expressing chitin synthase 1 (CHS-1) labeled with mCherryFP indicated only partial colocalization of the polarisome and the Spk core. N. crassa SPA-2-GFP was also found at the apex of forming branches but not in septa, suggesting that it participates only in areas of tip growth. A Deltaspa-2 strain displayed hyphae with uneven constrictions, apices with an unstable Spk, reduced growth rate and higher number of branches than the wild type strain, indicating that SPA-2 is required for the stability, behavior and morphology of the Spk and maintenance of regular apical growth in hyphae of N. crassa, although not for polarity or Spk establishment. PMID- 19281856 TI - The effects of shockwave on bone healing and systemic concentrations of nitric oxide (NO), TGF-beta1, VEGF and BMP-2 in long bone non-unions. AB - This study investigated the effects of extracorporeal shockwave treatment (ESWT) on bone healing and the systemic concentrations of nitric oxide (NO), TGF-beta1, VEGF and BMP-2 in long bone non-unions. Forty-two patients with 42 established non-unions of the femur and tibia were enrolled in this study. Each long bone non union was treated with 6000 impulses of shockwave at 28 kV in a single session. Ten milliliters of peripheral blood were obtained for measurements of serum NO level and osteogenic growth factors including TGF-beta1, VEGF and BMP-2; serum levels of calcium, alkaline phosphatase, calcitonin and parathyroid hormone before treatment and at 1 day, 1, 3 and 6 months after treatment. The evaluations for bone healing included clinical assessments and serial radiographic examinations. At 6 months, bony union was radiographically confirmed in 78.6%, and persistent non-union in 21.4%. Patients with bony union showed significantly higher serum NO level, TGF-beta1, VEGF and BMP-2 at 1 month after treatment as compared to patients with persistent non-union. Shockwave-promoted bone healing was associated with significant increases in serum NO level and osteogenic growth factors. The elevations of systemic concentration of NO level and the osteogenic factors may reflect a local stimulation of shockwave in bone healing in long bone non-unions. PMID- 19281857 TI - Microbial conversion of glucose to a novel chemical building block, 2-pyrone-4,6 dicarboxylic acid. AB - 2-Pyrone-4,6-dicarboxylic acid (PDC) is a catabolic intermediate in Sphingobium sp. SYK-6 (previously characterized as Sphingomonas paucimobilis SYK-6), which is a degrader of lignin-derived aromatic compounds. Recently, PDC has been also characterized as a novel starting material for several potentially useful synthetic polymers. In a previous study, we constructed a biosynthetic system in which PDC was generated efficiently from a chemically synthesized compound, protocatechuate. In order to develop an alternative system for production of PDC, we tried to generate it from glucose, which is a low-cost sugar that can be obtained from abundant cellulosic wastes and biomass crops. We designed a metabolic bypass to PDC from the shikimate pathway in recombinant Escherichia coli cells. PDC accumulated in the medium of recombinant E. coli cells that had been transformed with genes isolated from Emericella niger, E. coli, Pseudomonas putida, and Sphingobium sp. SYK-6. The yield of PDC depended on the combination of genes that we introduced into the cells and on the specific of host strain. Under optimal conditions, the yield and titer of PDC were, respectively, 17.3% and 0.35 mg/l when the concentration of glucose was 2 g/l and the culture volume was 50 ml. Our results open up the possibility of novel utilization of biomass as the source of a useful chemical building block. PMID- 19281858 TI - Health-related quality of life in patients with Barrett's esophagus: a systematic review. AB - BACKGROUND & AIMS: Barrett's esophagus (BE) affects approximately 10% of patients with chronic gastroesophageal reflux disease (GERD). Patients with BE are at risk for reduced health-related quality of life (HRQoL) associated with GERD, in addition to the potential psychosocial stress of carrying a diagnosis of a premalignant condition with a risk of esophageal adenocarcinoma. We sought to systematically review the published literature on HRQoL of patients with BE. METHODS: We searched PubMed, PsycINFO, and Cumulative Index to Nursing and Allied Health Literature for relevant clinical trials using a defined search strategy. We also manually searched relevant scientific meeting abstracts and related article bibliographies. Eligible articles were case series, cohort studies, or clinical trials that included one or more measures of HRQoL and/or quantitatively assessed burden of disease in patients with BE. Effect sizes were calculated when possible. RESULTS: Our initial search identified 95 articles. After 2 physician reviews, 25 articles met inclusion criteria. Data show that BE is associated with a significant decrement in HRQoL as measured by both generic and disease-targeted instruments. In addition, patients with BE are at risk for psychological consequences such as depression, anxiety, and stress, which may be related to their increased risk of esophageal adenocarcinoma. Compared with subjects with GERD alone or the general population, a diagnosis of BE also leads to increased health care use and spending. CONCLUSIONS: BE compromises multiple facets of patients' quality of life. Physicians and researchers should incorporate patient reported outcomes data including HRQoL measures when treating or studying patients with BE. PMID- 19281859 TI - A very low-carbohydrate diet improves symptoms and quality of life in diarrhea predominant irritable bowel syndrome. AB - BACKGROUND & AIMS: Patients with diarrhea-predominant irritable bowel syndrome (IBS-D) anecdotally report symptom improvement after initiating a very low carbohydrate diet (VLCD). This study prospectively evaluated a VLCD in IBS-D. METHODS: Participants with moderate to severe IBS-D were provided a 2-week standard diet, then 4 weeks of a VLCD (20 g carbohydrates/d). A responder was defined as having adequate relief of gastrointestinal symptoms for 2 or more weeks during the VLCD. Changes in abdominal pain, stool habits, and quality of life also were measured. RESULTS: Of the 17 participants enrolled, 13 completed the study and all met the responder definition, with 10 (77%) reporting adequate relief for all 4 VLCD weeks. Stool frequency decreased (2.6 +/- 0.8/d to 1.4 +/- 0.6/d; P < .001). Stool consistency improved from diarrheal to normal form (Bristol Stool Score, 5.3 +/- 0.7 to 3.8 +/- 1.2; P < .001). Pain scores and quality-of-life measures significantly improved. Outcomes were independent of weight loss. CONCLUSIONS: A VLCD provides adequate relief, and improves abdominal pain, stool habits, and quality of life in IBS-D. PMID- 19281860 TI - Incidence, prevalence, and clinical significance of abnormal hematologic indices in compensated cirrhosis. AB - BACKGROUND & AIMS: Patients with cirrhosis develop abnormal hematologic indices (HI) from multiple factors, including hypersplenism. We aimed to analyze the sequence of events and determine whether abnormal HI has prognostic significance. METHODS: We analyzed a database of 213 subjects with compensated cirrhosis without esophageal varices. Subjects were followed for approximately 9 years until the development of varices or variceal bleeding or completion of the study; 84 subjects developed varices. Abnormal HI was defined as anemia at baseline (hemoglobin, < or =13.5 g/dL for men and 11.5 g/dL for women), leukopenia (white blood cell counts, < or =4000/mm3), or thrombocytopenia (platelet counts, < or =150,000/mm3). The primary end points were death or transplant surgery. RESULTS: Most subjects had thrombocytopenia at baseline. Kaplan-Meier analysis showed that leukopenia occurred by 30 months (95% confidence interval, 18.5-53.6), and anemia occurred by 39.6 months (95% confidence interval, 24.1-49.9). Baseline thrombocytopenia (P = .0191) and leukopenia (P = .0383) were predictors of death or transplant, after adjusting for baseline hepatic venous pressure gradient (HVPG), and Child-Pugh scores. After a median of 5 years, a significant difference in death or transplant, mortality, and clinical decompensation was observed in patients who had leukopenia combined with thrombocytopenia at baseline compared with patients with normal HI (P < .0001). HVPG correlated with hemoglobin and white blood cell count (hemoglobin, r = -0.35, P < .0001; white blood cell count, r = -0.31, P < .0001). CONCLUSIONS: Thrombocytopenia is the most common and first abnormal HI to occur in patients with cirrhosis, followed by leukopenia and anemia. A combination of leukopenia and thrombocytopenia at baseline predicted increased morbidity and mortality. PMID- 19281861 TI - Persistent nausea and abdominal pain in a patient with delayed gastric emptying: shall we think of celiac disease? PMID- 19281862 TI - Features of duodenal cancer in patients with familial adenomatous polyposis. AB - BACKGROUND & AIMS: Most patients with familial adenomatous polyposis (FAP) develop duodenal adenomas; duodenal cancer is a major cause of mortality in this patient group. We reviewed cases of duodenal cancer in patients with FAP to identify factors that determine long-term cancer risk. METHODS: Twenty FAP patients (12 male) were identified from a registry database search. Data from registry and medical notes and endoscopic and histopathologic reports were evaluated. RESULTS: Of the cancers that developed in these patients, 11 were ampullary and 9 were duodenal. The median age at cancer diagnosis was 53 years. Seventeen patients died (median age at death, 57 y; median survival from diagnosis, 11 mo); the cause of death was metastatic or duodenal/ampullary cancer in 14 patients. Fifteen patients presented symptomatically (including 3 interval cancers while on surveillance). Two were diagnosed at surveillance and 3 were diagnosed during surgery performed for endoscopic features of advanced benign disease. Duodenal cancers were associated with a significantly lower mean colonic polyp count than ampullary cancers (496 +/- 282 vs 1322 +/- 735; P = .025); there appeared to be familial clustering of this cancer. When endoscopic data were available (n = 11 of 20), all ampullary cancers arose from ampullas greater than 1 cm. The Spigelman stage did not predict risk of ampullary cancer but did predict duodenal cancer (median stage 2 vs stage 4 for duodenal cancer). CONCLUSIONS: Once cancer arises in patients with FAP, prognosis is poor, so cancer prevention should be the main goal. Surveillance intervals should reflect both Spigelman staging and ampullary disease. PMID- 19281863 TI - An unusual cause of rectal bleeding. PMID- 19281864 TI - From hyperkalemia to ischemic colitis: a resinous way. PMID- 19281865 TI - Advanced colon polypectomy. PMID- 19281866 TI - Efficacy of esophageal impedance/pH monitoring in patients with refractory gastroesophageal reflux disease, on and off therapy. AB - BACKGROUND & AIMS: Intraluminal impedance monitoring has given new dimensions to the diagnosis of reflux disease. However, there is no defined algorithm for evaluating refractory reflux symptoms. We studied whether combined impedance/pH monitoring in patients on therapy can predict acid reflux in patients off therapy and whether testing should be carried out when patients are on or off therapy. METHODS: Thirty-nine adults (mean age, 50 years; 24 female) with refractory reflux symptoms were evaluated by impedance/pH monitoring while on therapy, followed by wireless pH monitoring while off therapy. Non-acid reflux events in patients on therapy were correlated with acid reflux parameters studied off therapy. In addition, the likelihood of test abnormalities on and off therapy was determined. RESULTS: In 25 of 39 patients (64%) on therapy, impedance testing was normal, with a median of 69 events (interquartile, 63.0-78.0). The percentage of time at pH <4 was within the normal range for all patients who were on therapy. The pH test results were abnormal in 28 of 39 patients (72%) when studied off therapy. Ninety-three of patients with abnormal impedance on therapy also had abnormal acid reflux off therapy. When both groups were off therapy, the patients with abnormal impedance parameters on therapy had significantly higher median (interquartile) 2-day baseline levels of esophageal acid exposure (8.7%, 6.9% 12.5%), compared with those of patients with normal impedance parameters while on therapy (6.0%, 2.8%-9.4%; P = .026). CONCLUSIONS: Abnormal impedance in patients on therapy predicts acid reflux in patients off therapy. In patients with refractory reflux, combined impedance/pH monitoring might provide the single best strategy for evaluation of reflux symptoms. PMID- 19281867 TI - Intraductal papillary mucinous neoplasm of the pancreas. PMID- 19281868 TI - Meckel's diverticulum diagnosed by double-balloon enteroscopy in an adult presenting with obscure gastrointestinal bleeding. PMID- 19281869 TI - Liver transplantation in patients with nonalcoholic steatohepatitis-related hepatocellular carcinoma. AB - BACKGROUND & AIMS: The increasing incidence of hepatocellular carcinoma in the United States is only partially accounted for by hepatitis C virus (HCV) infections. The prevalence of hepatocellular carcinoma in patients with nonalcoholic steatohepatitis (NASH) is not known; guidelines from the American Association for the Study of Liver Diseases do not recommend surveillance imaging. We sought to determine the prevalence of hepatocellular carcinoma among patients undergoing liver transplantation for NASH-related cirrhosis and their outcome after surgery, compared with controls. METHODS: We reviewed the records of adult patients with NASH cirrhosis who underwent liver transplantation by using a prospectively collected database from a single center. Data from patients with NASH cirrhosis were compared with matched controls who received transplantation for primary biliary cirrhosis/primary sclerosing cholangitis, alcoholic liver disease, or HCV. RESULTS: Seventeen of 98 patients (17%) with NASH cirrhosis were diagnosed with hepatocellular carcinoma. The mean age was 63 years, and 70% were male. Six patients were diagnosed with hepatocellular carcinoma incidentally on explant. Survival after liver transplantation was 88% after mean follow-up of 2.5 years. The number of NASH patients known to have hepatocellular carcinoma before liver transplantation was greater than the number of patients with primary biliary cirrhosis/primary sclerosing cholangitis and comparable to the number of patients with alcoholic liver disease and HCV. CONCLUSIONS: Patients with NASH cirrhosis are at risk for developing hepatocellular carcinoma; patients with NASH cirrhosis, especially men older than 50 years, should undergo surveillance imaging. Patients with NASH and hepatocellular carcinoma have good outcomes after liver transplantation. PMID- 19281870 TI - Cecal ameboma. PMID- 19281871 TI - A randomized controlled trial of acupuncture and moxibustion to treat Bell's palsy according to different stages: design and protocol. AB - Acupuncture to treat Bell's palsy is one of the most commonly used methods in China. There are a variety of acupuncture treatment options to treat Bell's palsy in clinical practice. Since Bell's palsy has three different path-stages (acute stage, resting stage and restoration stage), so whether acupuncture is effective in the different path-stages and which acupuncture treatment is the best method are major issues in acupuncture clinical trials about Bell's palsy. In this article, we report the design and protocol of a large sample multi-center randomized controlled trial to treat Bell's palsy with acupuncture. There are five acupuncture groups, with four according to different path-stages and one not. In total, 900 patients with Bell's palsy are enrolled in this study. These patients are randomly assigned to receive one of the following four treatment groups according to different path-stages, i.e. 1) staging acupuncture group, 2) staging acupuncture and moxibustion group, 3) staging electro-acupuncture group, 4) staging acupuncture along yangming musculature group or non-staging acupuncture control group. The outcome measurements in this trial are the effect comparison achieved among these five groups in terms of House-Brackmann scale (Global Score and Regional Score), Facial Disability Index scale, Classification scale of Facial Paralysis, and WHOQOL-BREF scale before randomization (baseline phase) and after randomization. The result of this trial will certify the efficacy of using staging acupuncture and moxibustion to treat Bell's palsy, and to approach a best acupuncture treatment among these five different methods for treating Bell's palsy. PMID- 19281872 TI - The n-hexane and chloroform fractions of Piper betle L. trigger different arms of immune responses in BALB/c mice and exhibit antifilarial activity against human lymphatic filarid Brugia malayi. AB - Modulation of immune functions by using herbal plants and their products has become fundamental regime of therapeutic approach. Piper betle Linn. (Piperaceae) is a widely distributed plant in the tropical and subtropical regions of the world and has been attributed as traditional herbal remedy for many diseases. We have recently reported the antifilarial and antileishmanial efficacy in the leaf extract of Bangla Mahoba landrace of P. betle which is a female plant. The present report describes the in vivo immunomodulatory efficacy of the crude methanolic extract and its n-hexane, chloroform, n-butanol fractions of the female plant at various dose levels ranging between 0.3 and 500 mg/kg in BALB/c. Attempts were also made to observe antifilarial activity of the active extracts and correlate it with the antigen specific immune responses in another rodent Mastomys coucha infected with human lymphatic filarial parasite Brugia malayi. The crude methanol extract and n-hexane fraction were found to potentiate significant (p<0.001) enhancement of both humoral (plaque forming cells, hemagglutination titre) as well as cell-mediated (lymphoproliferation, macrophage activation, delayed type hypersensitivity) immune responses in mice. The flow cytometric analysis of splenocytes of treated mice indicated enhanced population of T-cells (CD4(+), CD8(+)) and B-cells (CD19(+)). The n-hexane fraction (3 mg/kg) was found to induce biased type 2 cytokine response as revealed by increased IL-4(+) and decreased IFN-gamma(+) T-cell population while the chloroform fraction (10 mg/kg) produced a predominant type 1 cytokines. Crude methanolic extract (100 mg/kg) demonstrated a mixed type 1 and type 2 cytokine responses thus suggesting a remarkable immunomodulatory property in this plant. The induction of differential T-helper cell immune response appears ideal to overcome immunosuppression as observed in case of lymphatic, filarial Brugia malayi infection which may also be extended to other infections as well. PMID- 19281873 TI - Role of electrostatic repulsion on colloidal stability of Bacillus halmapalus alpha-amylase. AB - The colloidal stability of charged particles in suspension is often controlled by electrostatic repulsion, which can be rationalized in a semi-quantitative way by the DLVO theory. In the current study, we investigate this approach towards understanding irreversible protein aggregation, using Bacillus halmapalus alpha amylase (BHA) as a model protein. Repulsive forces between partly unfolded monomers were shown to strongly affect aggregation. Adding salt, increasing valence of counter ions or decreasing pH in the direction of pI resulted in a shift in the rate-limiting step from association to unfolding as evidenced by a change in aggregation kinetics from second to first-order in protein concentration. Charge screening effects by salts resulted in increased average size of protein aggregates but only moderately affected the secondary structure of protein within the aggregates. Salt and pH effects could be explained within the DLVO framework, indicating that partially unfolded BHA monomers can be modelled realistically as colloids with a random charge distribution. PMID- 19281874 TI - Crystal structures of T. vivax nucleoside hydrolase in complex with new potent and specific inhibitors. AB - Diseases caused by parasitic protozoa remain a major health problem, mainly due to old toxic drugs and rising drug resistance. Nucleoside hydrolases are key enzymes of the purine salvage pathway of parasites from the Trypanosomatidae family and are considered as possible drug targets. N-Arylmethyl substituted iminoribitols have been developed as selective nanomolar affinity inhibitors against the purine-specific nucleoside hydrolase of Trypanosoma vivax. The current paper describes the crystal structures of the T. vivax nucleoside hydrolase in complex with two of these inhibitors, to 1.3 and 1.85 A resolution. These high resolution structures provide an accurate picture of the mode of binding of these inhibitors and their mechanism of transition-state mimicry, and are valuable tools to guide further inhibitor design. Comparison of the current structures with previously solved structures of the enzyme in complex with ground state and transition-state-analogue inhibitors also allows for the elucidation of a detailed molecular mechanism of active-site loop opening/closing. These loop movements can be coupled to the complex kinetic mechanism of the T. vivax nucleoside hydrolase. PMID- 19281875 TI - Study of a sterol esterase secreted by Ophiostoma piceae: sequence, model and biochemical properties. AB - An extracellular sterol esterase from Ophiostoma piceae efficiently hydrolyzes sterol esters, triglycerides and p-nitrophenol esters. cDNA was screened with a probe obtained by PCR using as primers oligonucleotides corresponding to the N terminal and internal mature enzyme sequences and complete sequence was obtained by 3' rapid amplification of cDNA end (RACE) and inverse PCR. The O. piceae esterase gene had a length of 1.8 kbp and lacked introns. A search for proteins with related amino acid sequences revealed around 40% identity with lipases from Candida rugosa and Geotrichum candidum. Modelling the O. piceae enzyme, using the crystal structures of Lip1 and Lip3 from C. rugosa as templates, revealed a similar substrate-binding site, but some changes affecting the flap zone and the aromatic region of the tunnel may be responsible for the wide substrate specificity of this interesting sterol esterase. The ability of the new fungal esterase to hydrolyze triglycerides and esters of p-nitrophenol and cholesterol was compared with those of commercial lipases and cholesterol esterases showing the new enzyme the highest efficiency hydrolyzing triglycerides and sterol esters in the conditions assayed (in presence of Genapol X-100). Finally, the O. piceae gene was successfully expressed in Pichia pastoris, as a model organism to express fungal enzymes, resulting in higher levels of esterase activity than those obtained in the O. piceae cultures. In spite of its higher glycosylation degree, the recombinant enzyme was able to hydrolyze more efficiently than native enzyme the assayed substrates. PMID- 19281876 TI - Cloning and characterization of Lotus japonicus formate dehydrogenase: a possible correlation with hypoxia. AB - Formate dehydrogenases (FDHs, EC 1.2.1.2) comprise a group of enzymes found in both prokaryotes and eukaryotes that catalyse the oxidation of formate to CO(2). FDH1 from the model legume Lotus japonicus (LjFDH1) was cloned and expressed in E. coli BL21(DE3) as soluble active protein. The enzyme was purified using affinity chromatography on Cibacron blue 3GA-Sepharose. The enzymatic properties of the recombinant enzyme were investigated and the kinetic parameters (K(m), k(cat)) for a number of substrates were determined. Molecular modelling studies were also employed to create a model of LjFDH1, based on the known structure of the Pseudomonas sp. 101 enzyme. The molecular model was used to help interpret biochemical data concerning substrate specificity and catalytic mechanism of the enzyme. The temporal expression pattern of LjFDH1 gene was studied by real-time RT-PCR in various plant organs and during the development of nitrogen-fixing nodules. Furthermore, the spatial transcript accumulation during nodule development and in young seedpods was determined by in situ RNA-RNA hybridization. These results considered together indicate a possible role of formate oxidation by LjFDH1 in plant tissues characterized by relative hypoxia. PMID- 19281877 TI - Emotional aspects of chronic orofacial pain and surgical treatment. AB - OBJECTIVE: The aim of this pilot study was to investigate the psychological factors of chronic orofacial pain patients regarding hospitalization for surgical treatment. METHODS: We evaluated 30 patients (15 with temporomandibular disorder and 15 with trigeminal neuralgia) of three groups: 10 were hospitalized for surgery, 10 were newly diagnosed, and 10 had been clinically treated. Data were collected using a semi-structured interview and the Hospital Anxiety Depression Scale. RESULTS: Eighty percent reported lack of family support, 90% had important limitations in daily activities, and social aspects were the most affected (34%). Patients who were hospitalized for surgery had the highest degree of anxiety and expectation (90%; p<0.05). CONCLUSION: Surgery for chronic pain generates great expectations especially because it is considered a hope of cure. Clinically treated patients also might understand the factors associated to surgery choices and participate at the process of choosing. In general, chronic treatment for facial pain needs psychological support to cope with it. PMID- 19281878 TI - Sources of cholinergic input to the inferior colliculus. AB - We combined retrograde tracing with immunohistochemistry for choline acetyltransferase to identify the source of cholinergic input to the inferior colliculus (IC) in guinea pigs. Injection of a retrograde tracer into one IC labeled cells in many brainstem nuclei. Retrogradely-labeled cells that were also immunoreactive for choline acetyltransferase were identified in two nuclei in the midbrain tegmentum: the pedunculopontine tegmental nucleus (PPT) and the laterodorsal tegmental nucleus (LDT). More PPT and LDT cells project ipsilaterally than contralaterally to the IC and, on both sides, there are more projecting cells in the PPT than in the LDT. Double-labeled cells were not found in any other brainstem nucleus. A common feature of cholinergic cells in PPT and LDT is collateral projections to multiple targets. We placed different retrograde tracers into each IC to identify cells in PPT and LDT that project to both ICs. In both PPT and LDT, a substantial proportion (up to 57%) of the immunoreactive cells that contained tracer from the contralateral IC also contained tracer from the ipsilateral IC. We conclude that acetylcholine in the IC originates from the midbrain tegmental cholinergic nuclei: PPT and LDT. These nuclei are known to participate in arousal, the sleep/wake cycle and prepulse inhibition of acoustic startle. It is likely that the cholinergic input to the IC is directly associated with these functions. PMID- 19281879 TI - Composition-function relationships during IL-1-induced cartilage degradation and recovery. AB - OBJECTIVE: To examine the relationships between biochemical composition and mechanical properties of articular cartilage explants during interleukin-1 (IL-1) induced degradation and post-exposure recovery. DESIGN: Bovine articular cartilage explants were cultured for up to 32 days with or without 20 ng/mL IL-1. The dynamic shear modulus |G*(dyn)| and equilibrium and dynamic unconfined compression moduli (E(equil) and |E*(dyn)|) were measured at intervals throughout the culture period. In a subsequent recovery study, explants were cultured for 4 days with or without 20ng/mL IL-1 and for an additional 16 days in control media. The dynamic moduli |E*(dyn)| and |G*(dyn)| were measured at intervals during degeneration and recovery. Conditioned media and explant digests were assayed for sulfated glycosaminoglycans (sGAG) and collagen content. RESULTS: Continuous IL-1 stimulation triggered progressive decreases in E(equil), |E*(dyn)|, and |G*(dyn)| concomitant with the sequential release of sGAG and collagen from the explants. Brief IL-1 exposure resulted in a short release of sGAG but not collagen, followed by a gradual and incomplete repopulation of sGAG. The temporary sGAG depletion was associated with decreases in both |E*(dyn)| and |G*(dyn)| which also recovered after removal of IL-1. During IL-1-induced degradation and post exposure recovery, explant mechanical properties correlated well with tissue sGAG concentration. CONCLUSIONS: As previously shown for developing cartilages and engineered cartilage constructs, cytokine-induced changes in sGAG concentration (i.e., fixed charge density) are coincident with changes in compressive and shear properties of articular cartilage. Further, recovery of cartilage mechanical properties can be achieved by relief from proinflammatory stimuli and subsequent restoration of tissue sGAG concentration. PMID- 19281880 TI - An anatomical view on macrophages in tolerance. AB - Macrophages are divided into several functional subtypes involved in host defence, wound healing and immune regulation. In an alternate view, increased understanding of macrophage biology can also be gained by including anatomical location as a parameter. The focus of this review lies on tolerogenic features of macrophages relevant for autoimmune disease. The importance of macrophages as janitors clearing apoptotic cells and debris is seen in systemic lupus erythematosus (SLE) where a clearance defect often is connected to disease induction and progression. The apoptotic debris can be encountered by the macrophage in multiple anatomical compartments including the circulation but also in areas of intense cell death such as in the thymus and germinal center were tight regulation is important. PMID- 19281881 TI - How we say no: norepinephrine, inferior frontal gyrus, and response inhibition. PMID- 19281883 TI - Surgical clinics of North America. Foreword. PMID- 19281884 TI - The problem of infection has been inextricably linked with surgical therapy. Preface. PMID- 19281885 TI - Common microbial pathogens in surgical practice. AB - Despite ongoing major advances in antisepsis and in the development of potent antimicrobial agents since the early twentieth century, human beings remain subject to bacterial and fungal infection through mechanisms of virulence that continue to evade the latest advents in the microbiologic field today. Infection persists in surgical patients and only via the procurement of an in-depth knowledge of microorganism evolution and progression and an intricate understanding of human immune defense mechanisms are surgeons able to tackle infection in a fashion synonymous to that which allowed historic legends to transform the mere concept of surgery into reality. This article broadly describes current microbial pathogens and related issues in surgical disease. PMID- 19281887 TI - Antimicrobial agents for surgical infections. AB - Antimicrobial drugs are useful for the empiric and definitive treatment of infections in surgical patients. They are also important agents for perioperative antimicrobial prophylaxis. The proper selection and use of these drugs is a critical skill for surgeons. Although these agents have many beneficial effects, they also possess occasional adverse effects and should not be used indiscriminately. PMID- 19281886 TI - The stressed host response to infection: the disruptive signals and rhythms of systemic inflammation. AB - The cognate signals from sterile or pathogen-induced sources converge on the same recognition or response pathways. In the surgical patient, a systemic response to infection most often occurs in the context of ongoing inflammatory stress. Such an inflammatory response is modulated initially by the magnitude of injury and by patient-specific (endogenous) factors, such as confounding illness, age, and genetic variation. Over an extended period of stress, treatmentrelated (exogenous) factors add unpredictability to host responses to subsequent challenges, such as acquired infection. The host response is discussed in the context of how existing sterile stressors may modify the response to acquired infection in surgical patients. PMID- 19281888 TI - Adjunctive measures for treating surgical infections and sepsis. AB - The history of adjunctive treatments for severe sepsis has been fraught with more failures than successes. To date, there have been few interventions that have been demonstrated to be efficacious by multiple large, well-designed, multicenter randomized clinical trials. However, recent research into treatment strategies using drotrecogin alfa (activated), effective blood glucose management, early goal-directed therapy, protocolization of care, and intensivist management has demonstrated positive results. Further research is being conducted to verify the success of these initial trials. This article summarizes some of the available adjunctive treatments for severe sepsis. PMID- 19281889 TI - Prevention of surgical site infection. AB - Surgical site infections are a frequent cause of morbidity following surgical procedures. Gram-positive cocci, particularly staphylococci, cause many of these infections, although gram-negative organisms are also frequently involved. The risk of developing a surgical site infection is associated with a number of factors, including aspects of the operative procedure itself, such as wound classification, and patient-related variables, such as preexisting medical conditions. Both nonpharmacologic measures and antimicrobial prophylaxis for selected procedures are used to prevent development of these infections. Compliance with these generally accepted preventive principles may lead to overall decreases in the incidence of these infections. PMID- 19281890 TI - Prosthetic infection: lessons from treatment of the infected vascular graft. AB - Surgical prosthetics provide unquestioned benefit to patients in maintenance of life and limb. However, complications associated with prosthetic devices continue to represent a significant source of morbidity and mortality. Even as the surgeon becomes more adept at management of infections, the bacterial characteristics change in favor of increased virulence and greater resistance to antimicrobials. Excision or retention of the prosthesis depends on the time of presentation, the microbial isolates recovered, and the extent of surrounding tissue destruction. Recent work shows improving results with in situ replacement. PMID- 19281891 TI - Skin and soft tissue infections. AB - Skin and soft tissue infections are a common cause of hospitalization and use of antibiotic therapy, and may result in significant disability. Infections managed by surgeons may vary from simple, noncomplicated cellulitis to severe necrotizing soft tissue infections. The differentiation of necrotizing infections from nonnecrotizing infections is critical to achieving adequate surgical therapy. An understanding of the changing epidemiology of all complicated skin and soft tissue infections is required for selection of appropriate empiric antibiotic therapy. PMID- 19281892 TI - Intra-abdominal infections. AB - Most intra-abdominal infections develop from a source in the gastrointestinal tract. They are usually caused by aerobic and anaerobic enteric bacteria. Management generally involves an invasive procedure to control the source of the infection and antimicrobial therapy directed against the causative microorganisms. In a few highly select patients, these infections may be treated without a definitive source control procedure. Antimicrobial therapy is tailored to the individual patient, with narrower spectrum agents used to treat community acquired intraabdominal infections, and broader spectrum agents used for hospital acquired infections. Overall, these infections remain associated with significant morbidity and mortality, particularly in higher-risk patients who have impaired host defenses. PMID- 19281893 TI - Hospital-acquired pneumonia: pathophysiology, diagnosis, and treatment. AB - Hospital-acquired pneumonia (HAP) is one of the most common causes of nosocomial infection, morbidity, and mortality in hospitalized patients. Many patient- and disease-specific factors contribute to the pathophysiology of HAP, particularly in the surgical population. Risk-factor modification and inpatient prevention strategies can have a significant impact on the incidence of HAP. While the best diagnostic strategy remains a subject of some debate, prompt and appropriate antimicrobial therapy in patients suspected of having HAP has been shown to significantly decrease mortality. Because the pathogens responsible for HAP are frequently more virulent and have greater resistance to commonly used antimicrobials than other pathogens, clinicians must have knowledge of the resistance patterns at their institutions to choose appropriate therapy. PMID- 19281894 TI - Catheter-related bloodstream infection. AB - Catheter-related bloodstream infections (CR-BSIs) are a common, frequently preventable complication of central venous catheterization. CR-BSIs can be prevented by strict attention to insertion and maintenance of central venous catheters and removing unneeded catheters as soon as possible. Antiseptic- or antibiotic-impregnated catheters are also an effective tool to prevent infections. The diagnosis of CR-BSI is made largely based on culture results. CR BSIs should always be treated with antibiotics, and except in rare circumstances the infected catheter needs to be removed. PMID- 19281895 TI - Nosocomial urinary tract infection. AB - Nosocomial urinary tract infection, a common complication in surgical patients, is primarily related to the use of indwelling urinary catheters. Discontinuation of catheter usage within 2 days, whenever possible, is the cornerstone to avoiding these infections. Patients with asymptomatic bacteriuria may be treated with catheter removal only, and do not necessarily require antibiotic therapy. Patients with symptomatic infections should receive effective antimicrobial therapy, but removal of the catheter is also fundamental to clearing the urinary tract of infection. Antibiotic therapy of urinary tract infections is facilitated by the renal concentration of many antibiotics, permitting very high antibiotic concentrations to be achieved in the urine. PMID- 19281896 TI - Clostridium difficile colitis. AB - Clostridium difficile is the most common cause of infectious diarrhea in hospitalized patients. Its effects are mediated by C difficile toxins A and B. Recent outbreaks of severe colitis have been associated with a new strain of the bacterium that produces large amounts of the toxins. Although oral metronidazole and oral vancomycin can be used to treat C difficile-associated disease, intraluminal vancomycin is preferable for more severe C difficile colitis. Early surgical intervention can improve outcomes with fulminant colitis, although overall mortality remains high. PMID- 19281897 TI - Preventing bacterial resistance in surgical patients. AB - The development of antimicrobial resistant pathogens in surgical patients is a significant problem, and infections caused by these organisms are associated with increased morbidity and mortality. Programs to prevent the spread of resistant organisms emphasize standard infection control practices and appropriate antibiotic prescribing practices. Antibiotic restriction and selective reporting of bacterial susceptibilities have had limited success in decreasing development of resistance, and are difficult to maintain effectively in the absence of widespread clinician acceptance. Potentially more promising are integrated decision support tools, which can support optimal antibiotic selection while preserving the sense of clinician autonomy. The use of antibiotic cycling programs for critically ill patients may be another approach to preserving the efficacy of the currently antimicrobial against the continued pressure of increasing bacterial resistance. PMID- 19281898 TI - A systems approach to the prevention of surgical infections. AB - Infection after surgery continues to be a major source of morbidity and expense despite extensive efforts with educational programs, guidelines, and hospital based policies and procedures. The public and the government are demanding better performance and greater accountability. Our system operations within our institutions have failed. We need to adopt a culture dedicated to quality control through better information technology and data-driven initiatives to achieve improved clinical outcomes from infectious complications in surgery. PMID- 19281899 TI - Future diagnostic and therapeutic approaches in surgical infections. AB - Despite ongoing efforts to standardize therapy and improve management, the morbidity and mortality associated with surgical infections remain high. Continued innovation is required to improve outcomes further, particularly in the face of the increasing prevalence of multidrug resistant organisms. Although they remain in the experimental stages, a number of recent advances have the potential to have significant impact on the management and outcomes of surgical infections. These include novel diagnostic strategies, antimicrobials targeting microbial virulence factors, novel vaccines, and risk stratification based on genetic profiling. PMID- 19281900 TI - Occupational asthma: current concepts in pathogenesis, diagnosis, and management. AB - Occupational asthma (OA) may account for 25% or more of de novo adult asthma. The nomenclature has now better defined categories of OA caused by sensitizing agents and irritants, the latter best typified by the reactive airways dysfunction syndrome. Selecting the most appropriate diagnostic testing and management is driven by assessing whether a sensitizer is involved, and if so, identifying whether the sensitizing agent is a high-molecular-weight agent such as a protein or a low-molecular-weight reactive chemical such as an isocyanate. Increased understanding of the pathogenesis of OA from reactive chemical sensitizers is leading to development of better diagnostic testing and also an understanding of why testing for sensitization to such agents can be problematic. Risk factors for OA including possible genetic factors are being delineated better. Recently published guidelines for the diagnosis and management of occupational asthma are summarized; these reflect an increasingly robust evidence basis for recommendations. The utility of diagnostic tests for OA is being better defined by evidence, including sputum analysis performed in relation to work exposure with suspected sensitizers. Preventive and management approaches are reviewed. Longitudinal studies of patients with OA continue to show that timely removal from exposure leads to the best prognosis. PMID- 19281901 TI - Mechanisms of occupational asthma. AB - Inhalation of agents in the workplace can induce asthma in a relatively small proportion of exposed workers. Like nonoccupational asthma, occupational asthma is probably the result of multiple genetic, environmental, and behavioral influences. It is important that occupational asthma be recognized clinically because it has serious medical and socioeconomic consequences. Environmental factors that can affect the initiation of occupational asthma include the intrinsic characteristics of causative agents as well as the influence of the level and route of exposure at the workplace. The identification of host factors, polymorphisms, and candidate genes associated with occupational asthma may improve our understanding of mechanisms involved in asthma. High-molecular-weight compounds from biological sources and low-molecular-weight chemicals cause occupational asthma after a latent period of exposure. Although the clinical, functional, and pathologic features of occupational asthma caused by low molecular-weight agents resemble those of allergic asthma, the failure to detect specific IgE antibodies against most low-molecular-weight agents has resulted in a search for alternative or complementary physiopathologic mechanisms leading to airway sensitization. Recent advances have been made in the characterization of the immune response to low-molecular-weight agents. In contrast, the mechanism of the type of occupational asthma that occurs without latency after high-level exposure to irritants remains undetermined. PMID- 19281902 TI - Management of work-related asthma. AB - The physician managing work-related asthma (WRA) assumes many roles. The first is to confirm an accurate diagnosis, recognizing that WRA has multiple phenotypes, including sensitizer-induced occupational asthma (OA) caused by high-molecular weight (HMW) proteins or low-molecular-weight (LMW) chemicals; irritant-induced asthma; and work-exacerbated asthma. Pharmacotherapy for WRA is identical to nonwork-related asthma and should be guided by current asthma guidelines emphasizing control of both asthma impairment and risk domains. It is well established that the majority of workers diagnosed with OA caused by sensitizers experience persistent asthma after leaving the workplace. However, the long-term risk of persistent unremitting asthma can be prevented in a minority of cases, particularly with OA caused by LMW sensitizers, by establishing an early diagnosis of OA and reducing or eliminating exposure. The physician consultant may advise employers on workplace interventions needed to minimize effectively an affected employee's exposure to a causative agent or condition, and what measures are required to prevent new cases of WRA (ie, primary prevention). Although allergen immunotherapy has a putative role in treating and preventing WRA caused by HMW sensitizers, further study is needed. PMID- 19281903 TI - Current perspectives in occupational asthma. PMID- 19281904 TI - Advances in mechanisms of asthma, allergy, and immunology in 2008. AB - This review summarizes selected articles appearing in 2008 in the Journal. Articles chosen include those improving our understanding of mechanisms of allergic diseases by focusing on human basophil, mast cell, and eosinophil biology; IgE and its high-affinity receptor on various cells; novel properties of omalizumab; airways remodeling; and genetics. Articles from other journals have been included to supplement the topics presented. PMID- 19281905 TI - Advances in environmental and occupational disorders in 2008. AB - Substantial progress in understanding the role of environmental factors in allergic disease and asthma has been made in the past year. A number of new allergens have been described, and the impact of exposure to indoor allergens in the development of allergic respiratory disease is further confirmed. Exposures to environmental pollutants, particularly tobacco smoke in children, have furthered our knowledge of the detrimental effects of these exposures. This review highlights key advances in environmental and occupational exposures that contribute to the burden of allergic respiratory disease. PMID- 19281907 TI - Platelets play important roles in the late phase of the immediate hypersensitivity reaction. AB - BACKGROUND: Recent studies have shown that platelets have a role in most inflammatory reactions, but involvement of platelets in the immediate hypersensitivity reaction (IHR) in skin has not been examined. OBJECTIVE: To investigate the role of platelets in a mouse model of IgE-mediated IHR. METHODS: Mice were sensitized by injecting ovalbumin intraperitoneally and challenged by injecting ovalbumin intradermally into ears, with or without platelet depletion. RESULTS: Sensitized mice developed biphasic responses characterized by early phase and late-phase reactions (LPRs). Degranulation of mast cells in skin did not differ between platelet-depleted mice and controls. The early phase reaction was not suppressed at 1 hour, but platelet depletion significantly reduced the LPR at 24 hours (P < .01). Flow cytometry showed that P-selectin expression on platelets and the number of platelet-leukocyte aggregates were both higher in the blood of ovalbumin-challenged mice compared with sham-sensitized mice at 24 hours (P < .05). In platelet-depleted mice, the LPR was restored by infusing platelets from normal mice (P < .01). This effect did not occur by infusing platelets from P-selectin-deficient mice or by pretreating platelets with anti-P-selectin antibody. Injection of activated platelet supernatant into ears led to increased leukocyte infiltration at 24 hours, and this effect was blocked by pretreating the supernatant with several antichemokine antibodies. Systemic administration of antiplatelet compounds also suppressed the LPR significantly. CONCLUSION: These results show that platelets play important roles in the LPR of the IHR in skin by forming platelet-leukocyte complexes via P-selectin in blood and secreting several chemokines that attract leukocytes to skin. PMID- 19281908 TI - Roles of arginase variants, atopy, and ozone in childhood asthma. AB - BACKGROUND: Arginases (encoded by ARG1 and ARG2 genes) might play an important role in asthma pathogenesis through effects on nitrosative stress. Arginase expression is upregulated in asthma and varies with T(H)2 cytokine levels and oxidative stress. OBJECTIVE: We aimed to examine whether variants in these genes are associated with asthma and whether atopy and exposures to smoking and air pollution influence the associations. METHODS: Among non-Hispanic and Hispanic white participants of the Children's Health Study (n = 2946), we characterized variation in each locus (including promoter region) with 6 tag single nucleotide polymorphisms for ARG1 and 10 for ARG2. Asthma was defined by parental report of physician-diagnosed asthma at study entry. RESULTS: Both ARG1 and ARG2 genetic loci were significantly associated with asthma (global locus level P = .02 and .04, respectively). Compared with the most common haplotype within each locus, 1 ARG1 haplotype was associated with reduced risk (odds ratio [OR] per haplotype copy, 0.55; 95% CI, 0.36-0.84), and 1 ARG2 haplotype was associated with increased risk (OR per haplotype copy, 1.35; 95% CI, 1.04-1.76) of asthma. The effect of the ARG1 haplotype that was significantly associated with asthma varied by the child's history of atopy and ambient ozone (P(interaction) = .04 and .02, respectively). Among atopic children living in high-ozone communities, those carrying the ARG1 haplotype had reduced asthma risk (OR per haplotype copy, 0.12; 95% CI, 0.04-0.43; P(heterogeneity) across atopy/ozone categories = .008). CONCLUSIONS: ARG1 and ARG2 loci are associated with childhood asthma. The association between ARG1 variation and asthma might depend on atopy and ambient ozone levels. PMID- 19281909 TI - The histamine H4 receptor is functionally expressed on T(H)2 cells. AB - BACKGROUND: Histamine influences T-cell reactions via histamine receptors 1 and 2. The histamine receptor 4 (H(4)R) is the most recently identified histamine receptor and is also expressed on human CD4(+) T cells; however, its regulation and function are unclear. OBJECTIVE: To investigate expression, regulation, and function of the H(4)R on human CD4(+) T cells. METHODS: Histamine receptor 4 expression was studied by real-time quantitative RT-PCR and by flow cytometry. Effects of H(4)R stimulation on induction of the signal transduction molecules activator protein 1 (AP-1) and nuclear factor-kappaB (NF-kappaB) were determined by electrophoretic mobility shift assay and on cytokine production by RT-PCR and ELISA. RESULTS: Histamine receptor 4 mRNA and protein were expressed by CD4(+) T cells and upregulated by IL-4. Its expression was higher on T(H)2 cells than T(H)1 cells and naive T-cells. H(4)R agonists (clobenpropit and 4 methylhistamine) induced AP-1 in T(H)2 cells but not in T(H)1 cells. This effect was blocked by the H(4)R antagonist JNJ7777120. H(4)R agonists upregulated IL-31 mRNA in PBMCs and T(H)2 cells, a cytokine that has been associated with T(H)2 cells and the induction of pruritus. IL-31 mRNA induction by H(4)R stimulation was pronounced in PBMCs from patients with atopic dermatitis. Expression of IL-4, IL-5, and IL-13 was not altered by the H(4)R. CONCLUSION: Human CD4(+) T cells express a functional H(4)R. The receptor is upregulated under T(H)2 conditions, and its stimulation leads to induction of AP-1 and IL-31. PMID- 19281910 TI - Lung function test results in normal infants: a COPSAC sequel. PMID- 19281911 TI - Quantitative assessment of allergic shiners in children with allergic rhinitis. AB - BACKGROUND: The knowledge on allergic shiners is extremely limited. A conceivable tool able to quantify allergic shiners has not been established. OBJECTIVES: We sought to determine the significance and changeability of allergic shiners through our newly developed computerized method. METHODS: We developed a novel computerized method to measure allergic shiners and enrolled a cohort of children with or without allergic rhinitis. Children with allergic rhinitis were prospectively assessed. A standardized digital photograph was taken during each visit, and a modified Pediatric Rhinoconjunctivitis Quality of Life Questionnaire was completed. Subject global assessment for nose symptoms and subject global assessment for eye symptoms (SGAE) were self-recorded daily. RESULTS: We included 126 children with allergic rhinitis and 123 healthy control subjects. One hundred three (82%) participants with allergic rhinitis completed at least 4 prospective assessments. Shiners were darker (P < .001) and larger (P < .001) in children with allergic rhinitis. Darkness and sizes of allergic shiners were paradoxically inversely correlated (P = .02). Darkness of allergic shiners positively correlated with the duration of allergic rhinitis, practical problem scores, and SGAE values (P = .02, P = .004, and P = .002, respectively), but sizes of allergic shiners did not. Shiners were found to be darker in children with scores of eye symptoms of greater than 6, scores of practical problems of greater than 5, and SGAE values of greater than 0 (P = .02, P < .001, and P = .003, respectively), whereas shiners were larger in children with scores of other symptoms of greater than 9 and activity limitations of greater than 4 (P = .02 and P = .002, respectively). CONCLUSION: Computer-analyzed allergic shiners correlate with the chronicity and severity of allergic rhinitis. PMID- 19281912 TI - Assessing anaphylactic risk? Consider mast cell clonality. PMID- 19281913 TI - Angiotensin II receptor blockers in the treatment of the cardiovascular disease continuum. AB - BACKGROUND: The cardiovascular disease (CVD) continuum is a chain of events that begins with a host of risk factors, including dyslipidemia, hypertension, diabetes, visceral obesity, and smoking. If left untreated, it might progress to atherosclerosis, left ventricular hypertrophy, coronary artery disease, myocardial infarction, left ventricular remodeling, left ventricular enlargement, and eventually endstage heart disease and death. Initiation of treatment, at any stage in its course, might prevent or delay its further progression. OBJECTIVE: This review discusses data from doubleblind, randomized controlled trials (RCTs) that have investigated the effects of angiotensin II receptor blockers (ARBs) on various stages of the CVD continuum. METHODS: PubMed/MEDLINE was searched for relevant English-language double-blind RCTs using the years 1995 to 2008 and the terms angiotensin type II receptor blocker, renin-angiotensin system, hypertension, heart failure, left ventricular hypertropby, renal disease, stroke, and cerebrovascular disease. RESULTS: A total of 13 studies were included in this review. The results suggest that ARB-based therapy has cardioprotective, cerebroprotective, and renoprotective effects, including regression of left ventricular hypertrophy, reduction in the risk of stroke, and slowing of the progression of renal disease. CONCLUSIONS: Treatment of CVD risk factors, including hypertension, is increasingly recognized as a way to interrupt the CVD continuum. Activation of the renin-angiotensin system (RAS) contributes to the development and progression of CVD, and RAS blockade has been reported to be beneficial at all stages of the CVD continuum. PMID- 19281914 TI - Benefits and risks of clopidogrel use in patients with coronary artery disease: evidence from randomized studies and registries. AB - BACKGROUND: Aggressive antiplatelet treatment is currently an established practice in patients with coronary artery disease. OBJECTIVE: This article discusses the evidence from clinical trials and registries concerning the benefits and risks of clopidogrel in relation to the dose and timing of treatment and its use in particular patient populations (stratified by risk factors for major adverse cardiovascular events). METHODS: Human clinical trials, registries, and in vitro human platelet studies were identified through searches of MEDLINE, Scopus, Google Scholar, and ClinicalTrials.gov (January 1980-July 2008) using the terms clopidogrel in coronary artery disease, clopidogrel loading dose, clopidogrel pretreatment, and antiplatelet therapy for coronary artery disease. For inclusion in the review, publications had to compare clopidogrel administered at different doses and times across the spectrum of patients with coronary artery disease; present outcome data on mortality, reinfarction, stroke, or their combination; and report safety data pertaining to bleeding risk. RESULTS: The results of randomized trials support the benefit of administering a 300-mg loading dose of clopidogrel as part of both conservative and invasive strategies for the management of acute coronary syndromes (ACS). In a small randomized trial in patients with non-ST-segment elevation ACS, pretreatment with clopidogrel 600 mg >12 hours before coronary stenting was associated with significantly fewer major adverse cardiovascular events compared with pretreatment with clopidogrel 300 mg (P = 0.020). A randomized trial in patients with ACS undergoing percutaneous coronary intervention (PCI) reported a significant reduction in ischemic risk with prasugrel (60-mg loading dose, 10-mg/d maintenance dose) compared with clopidogrel (300-mg loading dose, 75-mg/d maintenance dose) (P < 0.001); however, the clinical benefit with prasugrel was accompanied by a significant increase in the risk of life-threatening bleeding (P = 0.01). Pretreatment with a 300-mg clopidogrel loading dose administered at least 15 hours before elective PCI was associated with a significant reduction in periprocedural ischemic events compared with 75 mg given after PCI (P = 0.028). In another randomized trial in patients undergoing elective PCI, pretreatment with clopidogrel 600 mg before diagnostic catheterization was associated with a significantly increased risk of minor bleeding complications (P = 0.027) without a significant benefit in terms of the primary ischemic end point. Data from registries support the use of a 300-mg loading dose of clopidogrel for the standard care of patients with ACS and indicate no significant benefit for pretreatment with loading doses >300 mg. One study of registry data found that pretreatment with clopidogrel 300 mg before elective PCI was associated with a significant reduction in major adverse cardiovascular events compared with a loading dose given immediately before or after the intervention (P = 0.002). CONCLUSIONS: Suppression of platelet activity through the use of antithrombotic agents should be balanced against the risk of atherothrombotic events. In patients with ACS, inhibition of platelet activation beyond that produced by clopidogrel 600 mg has been associated with an increased bleeding risk. In patients undergoing elective PCI, pretreatment with clopidogrel 300 mg administered >15 hours before the procedure achieved the optimal risk-benefit ratio. In cases of unplanned PCI, administration of clopidogrel 600 mg immediately after diagnostic catheterization was well tolerated and effective. PMID- 19281915 TI - Cost-effectiveness of intensive atorvastatin treatment in high-risk patients compared with usual care in a postgeneric statin market: economic analysis of the Aggressive Lipid-lowering Initiation Abates New Cardiac Events (ALLIANCE) study. AB - BACKGROUND: Intensive lipid-lowering therapy reduces cardiovascular (CV) event rates more than moderate therapy. Currently available generic statins cost less but provide less risk reduction. OBJECTIVE: Our goal was to calculate the incremental cost-effectiveness ratio (ICER) comparing an intensive atorvastatin based regimen versus usual care, defined as prevalent statin therapy used at the time of the Aggressive Lipid-Lowering Initiation Abates New Cardiac Events (ALLIANCE) study-a trial of usual community practice that was conducted among patients enrolled in managed-care plans. METHODS: A 7-year cost model incorporated patient-level CV event rates and mortality data from the ALLIANCE study. To address the managed-care perspective, costs for clinical end points were estimated from administrative claims data for equivalent events. Direct medical costs included drugs, acute events, and a maximum of 36 months of follow up costs. Average wholesale pricing was used for drug costs. Differences in patient copayments were incorporated and a 3 % discount factor was applied. Scenario analyses explored 3- and 5-year time frames, varying discount rates, drug costs, and copayment assumptions. RESULTS: The baseline ICER using all events examined in the ALLIANCE study and a 7-year time frame was US $10,344 (year-2007 value) per event avoided. Over 7 years, the number needed to treat to avoid a clinical end point was 11 patients. Using a realworld economic model that considered atorvastatin becoming generic in 3 years, starting patients on branded and converting them to generic atorvastatin would be a cost-saving strategy compared with current generic statin use. CONCLUSIONS: Intensive lipid-lowering using atorvastatin increased prescription drug costs of treating CV disease patients but reduced CV events. Medical cost offsets associated with reduced events, based on data from the ALLIANCE study, resulted in ICERs within accepted cost-effectiveness ranges comparing atorvastatin with a generic statin. Atorvastatin-based regimens produced cost savings from a managed-care perspective when the anticipated impact of the generic availability of atorvastatin was modeled. PMID- 19281916 TI - Retrospective claims database analysis to determine relationship between renin angiotensin system agents, rehospitalization, and health care costs in patients with heart failure or myocardial infarction. AB - BACKGROUND: Heart failure (HF) and myocardial infarction (MI) cause considerable morbidity and mortality, but the outcomes and health care costs related to adherence to treatment guidelines for HF and MI are not fully understood. OBJECTIVES: The aims of this study were as follows: (1) to determine the proportion of patients discharged from the hospital with a primary diagnosis of HF or MI who subsequently received prescriptions for American Heart Association/American College of Cardiology-recommended angiotensin-converting enzyme inhibitors (ACEIs) or angiotensin receptor blockers (ARBs), also referred to as renin-angiotensin system agents; (2) to investigate the relationship between adherence to and persistence with ACEIs/ARBs and risk of rehospitalization; and (3) to assess the relationship between adherence to and persistence with ACEIs or ARBs, cardiovascular-related health care costs, and total health care costs. METHODS: Using the prescription and medical service records of a large national pharmacy-benefit database, we conducted a retrospective analysis of patients discharged from the hospital with a primary diagnosis of HF or MI between July 1, 2003, and June 30, 2006. Medication adherence, persistence, rehospitalization risk, and health care costs during 1 year follow-up were measured. Logistic regression models were used to estimate the likelihood of rehospitalization for different levels of adherence and persistence. Generalized linear models were used to investigate the impact of adherence and persistence on total health care costs and cardiovascular-related health care costs. RESULTS: A total of 799 HF and 696 MI patients were included in the analysis; 57.20% of HF patients and 59.20% of MI patients were prescribed an ACEI or an ARB after discharge from the hospital. The mean (SD) age was 65.7 (13.7) years in the HF group and 60.6 (10.7) years in the MI group. In both groups, men accounted for a greater proportion of the patients than women. In the HF group, adherence and persistence were associated with a lower likelihood of rehospitalization compared with nonadherence and nonpersistence (P < 0.042 and P < 0.005, respectively). In the MI group, there was no significant difference in rehospitalization risk between those who were adherent and those who were not. However, among these patients, persistence was associated with a lower risk of rehospitalization than nonpersistence (P < 0.036). Adherence and persistence were associated with lower health care costs (HF: P < 0.001 for all comparisons; MI: P < 0.019 for adherence and total cost, P = NS for persistence and total cost, P < 0.012 for adherence and cardiovascular cost, P < 0.031 for persistence and cardiovascular cost). CONCLUSION: Adherence to and persistence with ACEIs or ARBs may reduce the risk of rehospitalization in patients with HF or MI, thereby potentially reducing health care costs. PMID- 19281917 TI - Long-term persistence with statin therapy: a nationwide register study in Finland. AB - BACKGROUND: Preventive statin therapy is often recommended as lifelong treatment. OBJECTIVE: The aim of this study was to analyze persistence with statin therapy over a decade of use and to identify factors associated with its discontinuation. METHODS: Persistence with therapy among new users of statins in 1995 was followed up until December 31, 2005, in Finland using the nationwide drug reimbursement register. Cumulative persistence was analyzed using Kaplan-Meier analysis. A Cox regression model was applied to analyze associations of various baseline covariates with discontinuation. We further modeled the association of time specific covariates by stratifying the duration of therapy in years and using a logistic regression in which those continuing therapy until the end of follow-up (persistent users) formed the reference group. Adherence, defined as the proportion of days covered by statins, stratified by the timing of discontinuation, was computed for the respective groups. RESULTS: Of the 18,072 new statin users, 73.3% (n =13,254) were aged >54 years and 54.8% (n =9908) were men. Of this cohort, 43.9% (n = 7926) were using statins throughout and at the end of the tenth year. Sex was not associated with persistence at any point. In the Cox model, persons aged 45 to 74 years at initiation were more likely to continue statin use than younger or older age groups. Among those who still used statins after the fifth year of observation, the age difference was not observed in the logistic regression model. The use of 1, 2, 3, or > or =4 cardiovascular drugs before the initiation predicted continuation relative to no cardiovascular drug use (hazard ratio for discontinuation significantly <1.00 in all comparisons). Adherence was best (median 93.9%) among the persistent users. CONCLUSIONS: The 10-year persistence with statin use in this general population was approximately 44%. Persons aged 45 to 74 years at initiation and those with at least 1 prescription for another cardiovascular medication were the most likely to continue statin therapy up to the fifth year. PMID- 19281918 TI - Prescribing of beta-blockers after myocardial infarction: a preliminary study of physician motivations and barriers. AB - BACKGROUND: Clinical practice guidelines (CPGs) recommend beta-blocker medications in the management of post-myocardial infarction (MI) patients to prevent subsequent MI; evidence suggests that this therapy continues to be underutilized. The decision-making process that physicians use for treating post MI patients should be explored; behavior-change interventions have been proposed among researchers in this field. OBJECTIVE: The aim of this pilot study was to develop measures for examining this issue using the transtheoretical model (TTM) of change, including physician stage of readiness for change and physician salience (ie, decisional balance) for the pros (ie, facilitators) and cons (ie, barriers) of prescribing beta-blockers to post-MI patients. METHODS: In this cross-sectional study, a TTM-based questionnaire was developed from the literature and from interviews with a convenience sample of physicians practicing in cardiology; the questionnaire was subjected to expert panels for revision and was pretested. It was then mailed to all physicians practicing family medicine, internal medicine, or cardiology during the autumn of 2003 in northern West Virginia, a state with a high prevalence of cardiovascular disease and MI-related mortality. RESULTS: Among the 309 physicians who received the questionnaire, 103 responded; 55 responses were complete and eligible for inclusion in the analysis. Most (92.7%) responded to the TTM measures in the action and maintenance stages of change for prescribing beta-blockers for post-MI patients. Results also suggested they had significant salience (P < 0.05) for the cons of prescribing beta-blocker therapy to patients with relative contraindications that were indicated in the earlier versions of the CPGs, but not in the latest version. Other specific barriers to prescribing were also identified. CONCLUSIONS: Results of this preliminary study suggest that it would be relevant to increase physician knowledge of the updated CPGs regarding use of beta-blockers in the presence of conditions previously deemed relative contraindications. PMID- 19281919 TI - Heart failure with normal left ventricular ejection fraction. AB - It is estimated that approximately 50% of the heart failure population has a normal left ventricular ejection fraction, a complex broadly referred to as heart failure with normal left ventricular ejection fraction (HFNEF). While these patients have been considered in epidemiologic studies and clinical trials to represent a single pool of patients, limited more detailed studies indicate that HFNEF patients are a very heterogeneous group, with a number of key pathophysiologic mechanisms. This review summarizes and critically analyzes available data on the pathophysiology of HFNEF, placing it into context with a recently developed diagnostic algorithm. We evaluate the utility of commonly applied echocardiographic measures and biomarkers and integrate mechanistic observations into potential future therapeutic directions. PMID- 19281920 TI - A randomized controlled trial of a paclitaxel-eluting stent versus a similar bare metal stent in saphenous vein graft lesions the SOS (Stenting of Saphenous Vein Grafts) trial. AB - OBJECTIVES: The aim of this study was to compare the frequency of angiographic restenosis and clinical events between a paclitaxel-eluting stent (PES) and a similar bare-metal stent (BMS) in saphenous vein graft (SVG) lesions. BACKGROUND: There are conflicting and mostly retrospective data on outcomes after drug eluting stent implantation in SVGs. METHODS: Patients requiring SVG lesion stenting were randomized to BMS or PES. The primary study end point was binary in segment restenosis at 12-month follow-up quantitative coronary angiography. Secondary end points included death, myocardial infarction, ischemia-driven target vessel and lesion revascularization, and target vessel failure. RESULTS: Eighty patients with 112 lesions in 88 SVGs were randomized to a BMS (39 patients, 43 grafts, 55 lesions) or PES (41 patients, 45 grafts, 57 lesions). Binary angiographic restenosis occurred in 51% of the BMS-treated lesions versus 9% of the PES-treated lesions (relative risk: 0.18; 95% confidence interval [CI]: 0.07 to 0.48, p < 0.0001). During a median follow-up of 1.5 years the PES patients had less target lesion revascularization (28% vs. 5%, hazard ratio: 0.38; 95% CI: 0.15 to 0.74, p = 0.003) and target vessel failure (46% vs. 22%, hazard ratio: 0.65; 95% CI: 0.42 to 0.96, p = 0.03), a trend toward less target vessel revascularization (31% vs. 15%, hazard ratio: 0.66; 95% CI: 0.39 to 1.05, p = 0.08) and myocardial infarction (31% vs. 15%, hazard ratio: 0.67; 95% CI: 0.40 to 1.08, p = 0.10), and similar mortality (5% vs. 12%, hazard ratio: 1.56; 95% CI: 0.72 to 4.11, p = 0.27). CONCLUSIONS: In SVG lesions, PES are associated with lower rates of angiographic restenosis and target vessel failure than BMS. PMID- 19281921 TI - Drug-eluting stents for saphenous vein graft lesions: the limits of evidence. PMID- 19281922 TI - Prevalence of low low-density lipoprotein cholesterol with elevated high sensitivity C-reactive protein in the U.S.: implications of the JUPITER (Justification for the Use of Statins in Primary Prevention: An Intervention Trial Evaluating Rosuvastatin) study. AB - OBJECTIVES: We assessed the prevalence of low-density lipoprotein-cholesterol (LDL-C) <130 mg/dl with elevated high-sensitivity C-reactive protein (hsCRP) in the National Health And Nutrition Examination Survey (NHANES), weighted to be representative of the general U.S. population. BACKGROUND: Rosuvastatin therapy in the JUPITER (Justification for the Use of Statins in Primary Prevention: An Intervention Trial Evaluating Rosuvastatin) study reduced cardiovascular events among older adults with LDL-C <130 mg/dl and hsCRP > or =2 mg/l. METHODS: Using 1999 to 2004 NHANES data, we categorized men age > or =50 years and women age > or =60 years by fasting LDL-C and hsCRP levels, excluding individuals with prevalent coronary heart disease, coronary heart disease equivalent (including diabetes), and other JUPITER exclusions. RESULTS: A total of 3.9 million men age > or =50 years and 2.6 million women age > or =60 years meeting JUPITER eligibility criteria had fasting LDL-C <130 mg/dl and hsCRP > or =2 mg/l. In addition, 6.7 million older adults with elevated hsCRP > or =2 mg/l have LDL-C levels that exceed their National Cholesterol Education Program goals. CONCLUSIONS: Extrapolating JUPITER eligibility to NHANES, an estimated 6.5 million additional adults could be potential candidates to initiate statin therapy. PMID- 19281923 TI - The association of anger and hostility with future coronary heart disease: a meta analytic review of prospective evidence. AB - OBJECTIVES: This review aimed to evaluate the association between anger and hostility and coronary heart disease (CHD) in prospective cohort studies using quantitative methods. BACKGROUND: The harmful effect of anger and hostility on CHD has been widely asserted, but previous reviews have been inconclusive. METHODS: We searched general bibliographic databases: MEDLINE, PsycINFO, Web of Science, and PubMed up to November 2008. Two reviewers independently extracted data on study characteristics, quality, and estimates of associations. RESULTS: There were 25 studies (21 articles) investigating CHD outcomes in initially healthy populations and 19 studies (18 articles) of samples with existing CHD. Anger and hostility were associated with increased CHD events in the healthy population studies (combined hazard ratio [HR]: 1.19; 95% confidence interval [CI]: 1.05 to 1.35, p = 0.008) and with poor prognosis in the CHD population studies (HR: 1.24; 95% CI: 1.08 to 1.42, p = 0.002). There were indications of publication bias in these reports, although the fail-safe numbers were 2,020 and 750 for healthy and disease population studies, respectively. Intriguingly, the harmful effect of anger and hostility on CHD events in the healthy populations was greater in men than women. In studies of participants with CHD at baseline that controlled fully for basal disease status and treatment, the association of anger and hostility with poor prognosis persisted. CONCLUSIONS: The current review suggests that anger and hostility are associated with CHD outcomes both in healthy and CHD populations. Besides conventional physical and pharmacological interventions, this supports the use of psychological management focusing on anger and hostility in the prevention and treatment of CHD. PMID- 19281924 TI - Anger, depression, and anxiety in cardiac patients: the complexity of individual differences in psychological risk. PMID- 19281925 TI - Depression and risk of sudden cardiac death and coronary heart disease in women: results from the Nurses' Health Study. AB - OBJECTIVES: We assessed the association between depression and sudden cardiac death (SCD) and cardiac events among individuals without baseline coronary heart disease (CHD). BACKGROUND: Depression is a risk factor for cardiac events and mortality among those with CHD, possibly from arrhythmia. METHODS: We studied depressive symptoms and a proxy variable for clinical depression consisting of severe symptoms and/or antidepressant medication use and their relationship to cardiac events in the Nurses' Health Study. Questionnaires in 1992, 1996, and 2000 assessed symptoms with the Mental Health Index (MHI-5), and antidepressant use was assessed in 1996 and 2000. Primary end points included SCD, fatal CHD, and nonfatal myocardial infarction. RESULTS: Among 63,469 women without prior CHD/stroke in 1992, 7.9% had MHI-5 scores <53, previously found to predict clinical depression. Depressive symptoms were associated with CHD events, and the relationship was strongest for fatal CHD, where the association remained significant even after controlling for CHD risk factors (hazard ratio [HR]: 1.49; 95% confidence interval [CI]: 1.11 to 2.00 for MHI-5 score <53). In models from 1996 onward, our proxy variable for clinical depression was most associated with SCD in multivariable models (HR: 2.33, 95% CI: 1.47 to 3.70), and this risk was primarily due to a specific relationship between antidepressant use and SCD (HR: 3.34, 95% CI: 2.03 to 5.50). CONCLUSIONS: In this cohort of women without baseline CHD, depressive symptoms were associated with fatal CHD, and a measure of clinical depression including antidepressant use was specifically associated with SCD. Although antidepressant use might be a marker of worse depression, its specific association with SCD merits further study. PMID- 19281926 TI - Do depression or antidepressants increase cardiovascular mortality? The absence of proof might be more important than the proof of absence. PMID- 19281927 TI - Reconstituted high-density lipoprotein increases plasma high-density lipoprotein anti-inflammatory properties and cholesterol efflux capacity in patients with type 2 diabetes. AB - OBJECTIVES: Our aim was to investigate the effects of reconstituted high-density lipoprotein (rHDL) infusions on plasma high-density lipoprotein (HDL) anti inflammatory properties and ex vivo cholesterol efflux in patients with type 2 diabetes. BACKGROUND: The anti-inflammatory effects of HDL contribute to protection from cardiovascular events. Individuals with type 2 diabetes are at elevated risk for cardiovascular disease, and typically have low HDL with reduced anti-inflammatory properties. METHODS: Thirteen fasting male patients (mean age 52 years) with type 2 diabetes mellitus received both rHDL (80 mg/kg of apolipoprotein A-I) and a saline placebo on separate occasions in a randomized cross-over design study. Changes in the ability of isolated HDL to influence the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in human coronary artery endothelial cells was the main outcome measure. Other outcome measures included expression of the key integrin, CD11b on patient monocytes, adhesiveness of patient neutrophils to fibrinogen, and the ability of plasma to promote cholesterol efflux to THP-1 macrophages. RESULTS: Four and 72 h post-rHDL infusion, the anti-inflammatory properties of isolated HDL increased in parallel to their concentration in plasma (by up to 25%, p < 0.01). Participants' peripheral blood monocyte CD11b expression and neutrophil adhesion to a fibrinogen matrix was also reduced 72 h post-rHDL, compared with that seen in placebo (p = 0.02). rHDL increased the capacity of plasma to receive cholesterol from THP-1 macrophages by 1 h up to 72 h post-infusion (by 40% to 60%, p < 0.05). CONCLUSIONS: rHDL infusions have significant, potentially atheroprotective effects in individuals with diabetes, including suppression of inflammation and enhancement of cholesterol efflux. PMID- 19281928 TI - An armored heart in constrictive pericarditis. PMID- 19281929 TI - Comparative effectiveness research. PMID- 19281930 TI - AHA/ACCF/HRS recommendations for the standardization and interpretation of the electrocardiogram: part III: intraventricular conduction disturbances: a scientific statement from the American Heart Association Electrocardiography and Arrhythmias Committee, Council on Clinical Cardiology; the American College of Cardiology Foundation; and the Heart Rhythm Society. Endorsed by the International Society for Computerized Electrocardiology. PMID- 19281931 TI - AHA/ACCF/HRS recommendations for the standardization and interpretation of the electrocardiogram: part IV: the ST segment, T and U waves, and the QT interval: a scientific statement from the American Heart Association Electrocardiography and Arrhythmias Committee, Council on Clinical Cardiology; the American College of Cardiology Foundation; and the Heart Rhythm Society. Endorsed by the International Society for Computerized Electrocardiology. PMID- 19281932 TI - AHA/ACCF/HRS recommendations for the standardization and interpretation of the electrocardiogram: part V: electrocardiogram changes associated with cardiac chamber hypertrophy: a scientific statement from the American Heart Association Electrocardiography and Arrhythmias Committee, Council on Clinical Cardiology; the American College of Cardiology Foundation; and the Heart Rhythm Society. Endorsed by the International Society for Computerized Electrocardiology. PMID- 19281933 TI - AHA/ACCF/HRS recommendations for the standardization and interpretation of the electrocardiogram: part VI: acute ischemia/infarction: a scientific statement from the American Heart Association Electrocardiography and Arrhythmias Committee, Council on Clinical Cardiology; the American College of Cardiology Foundation; and the Heart Rhythm Society. Endorsed by the International Society for Computerized Electrocardiology. PMID- 19281934 TI - Intensive care or merely therapy? PMID- 19281935 TI - Efficacy and safety profile of a single dose of hydromorphone compared with morphine in older adults with acute, severe pain: a prospective, randomized, double-blind clinical trial. AB - BACKGROUND: Older adults (ie, those aged > or =65 years) are the fastest growing segment of the US population, with an estimated approximately 71 million expected by 2030. Over the past 10 years, there has been an 11% increase in the number of emergency department (ED) visits by older adults, and pain is their most common chief complaint. OBJECTIVE: The goal of this study was to compare weight-based IV hydromorphone and IV morphine in adults aged > or =65 years presenting to the ED with acute, severe pain. METHODS: This was a prospective, randomized, double blind clinical trial of older adults with acute, severe pain at an adult, urban academic ED. Patients were randomly allocated to receive a single dose of 0.0075 mg/kg IV hydromorphone or 0.05-mg/kg IV morphine. The primary outcome was the between-group difference in decrease in pain from baseline to 30 minutes after the medications were infused. Patients' degree of pain was measured on a numerical rating scale (NRS) where "0" was defined as "no pain" and "10" was defined as "the worst pain possible." Adverse effects, pain reduction at 10 minutes and 2 hours postbaseline, patient evaluations of satisfaction and pain relief at 30 minutes postbaseline, and use of additional analgesics and antiemetics were tracked as secondary outcomes. RESULTS: A total of 194 patients were randomized to treatment; 183 patients (hydromorphone group, n = 93; morphine group, n = 90 [overall mean (SD) age, 75 (8) years]) had sufficient data for analysis at the primary end point of 30 minutes postbaseline. The mean decrease in pain from baseline to 30 minutes in patients allocated to IV hydromorphone was 3.8 versus 3.3 NRS units in patients allocated to IV morphine. This difference of 0.5 NRS unit (95% CI, -0.2 to 1.3) was neither clinically nor statistically significant. A majority of patients in both groups (57.0% randomized to hydromorphone and 58.9% randomized to morphine) failed to achieve a > or =50% reduction in pain within 30 minutes of treatment. The incidence of adverse effects from baseline to 30 minutes was not statistically different in the 2 groups. CONCLUSIONS: A single dose of IV hydromorphone at 0.0075 mg/kg was neither clinically nor statistically different from IV morphine at 0.05 mg/kg for the treatment of acute, severe pain at 30 minutes postbaseline in these older adults in the ED. The incidence of adverse effects was not statistically different. Our data suggest that hydromorphone and morphine in the doses given had similar efficacy and safety profiles in these older adults. Neither regimen provided > or =50% pain relief for the majority of patients. Future investigations of acute pain management in older adults should examine the efficacy and safety of higher initial (loading) doses of opioids titrated at frequent intervals until adequate analgesia is achieved. PMID- 19281936 TI - Effect of a retrospective drug utilization review on potentially inappropriate prescribing in the elderly. AB - BACKGROUND: Use of potentially inappropriate medications or drugs to be avoided in the elderly (DAE) continues to be widespread. Although the literature suggests DAE are associated with negative health outcomes, educational interventions have had a positive impact on inappropriate prescribing. OBJECTIVES: The objectives of this study were to identify those members aged > or =65 years participating in a Medicare Part D Blue Cross and Blue Shield (BCBS) benefit plan who were receiving medications that may be inappropriate for use in older adults and, through a retrospective drug utilization review (RetroDUR), to notify their prcscribers of the possible safety concerns with continued use. METHODS: The analysis used retrospective administrative pharmacy claims data from 3 Medicare Part D BCBS plans across 4 states. Plan members aged > or =65 years who had a claim for > or =1 DAE during a 30-day review period (August 15, 2007-September 14, 2007) with a minimum supply of 7 days of medication were identified. The National Committee for Quality Assurance 2007 Healthcare Effectiveness Data and Information Set measures for Medicare were used to determine DAE. A packet of information was mailed to prescribers identifying patients who had a claim for > or =1 DAE. Members were then assessed for the presence of a drug in the same drug class 6 months after the initial analysis. RESULTS: Of a possible 328,000 eligible members, 16,973 (5.2%) had a claim for > or =1 DAE during the 30-day review period. A total of 7963 intervention prescriber letters were mailed, affecting 13,198 members with 14,267 DAE claims. The final analyzable intervention cohort consisted of 10,364 members with 11,062 DAE claims. Overall, 5403 claims (48.8%) for DAE were defined as discontinued after 6 months. The most common DAE in the study were estrogens, propoxyphene, muscle relaxants, anticholinergics, antihistamines, and nitrofurantoin, accounting for 9682 claims (87.5%). At the 6 month follow-up, reductions in claims for each of the top 6 drug/drug classes ranged from 31.3% to 66.7%. As a class, the anticholinergics had the highest rate of discontinuation. CONCLUSIONS: The DAE RetroDUR was associated with a possible reduction in the use of potentially inappropriate prescription medications in these older adults. Further research, using a control population, is needed to show the impact on health care utilization and costs, adverse drug events, and health care and quality-of-life outcomes. PMID- 19281937 TI - Impact of a geriatric nursing home palliative care service on unnecessary medication prescribing. AB - BACKGROUND: There is a lack of studies concerning improvement of medication use in palliative care patients in nursing homes. OBJECTIVE: This study was conducted to evaluate whether a geriatric palliative care team reduced unnecessary medication prescribing for elderly veterans residing in a nursing home. METHODS: This was a retrospective, descriptive study of patients who died while residing in a geriatric palliative care unit between August 1, 2005, and July 31, 2007. Prescribed medications were evaluated using the Unnecessary Drug Use Measure, which contains 3 items from the Medication Appropriateness Index concerning lack of indication, lack of effectiveness, and therapeutic duplication. This measure was applied at 2 time points: on transfer/admission to the palliative care unit and at the last 30-day pharmacist medication review before death. Paired t tests and McNemar tests were used to compare medication use at these 2 points. RESULTS: Eighty-nine patients were included in the study. The majority were male (97.8%) and white (78.7%), with a mean (SD) age of 79.7 (7.8) years. The median length of stay on the unit was 39.0 days, and the mean number of chronic medical conditions was 8.4 (4.3). At baseline, the mean number of scheduled medications was 9.7 (4.3). The number of unnecessary medications per patient decreased from a mean of 1.7 (1.5) at admission to 0.6 (0.8) at closeout (P = 0.003). The decrease was seen in all 3 categories of the Unnecessary Drug Use Measure. CONCLUSIONS: The geriatric palliative care team was associated with a reduction in the number of unnecessary medications prescribed for older veterans in this nursing home. Future studies should evaluate the impact of decreasing unnecessary prescribing on clinical outcomes such as adverse drug reactions. PMID- 19281938 TI - Effect of concomitant administration of meropenem and valproic acid in an elderly Chinese patient. AB - BACKGROUND: Meropenem is a carbapenem with a broad spectrum of activity against beta-lactamase-producing organisms. Valproic acid is widely used in the treatment of generalized tonic-clonic and partial seizures. Concomitant administration of meropenem and valproic acid reportedly leads to a rapid decline in serum concentrations of valproic acid, which is sometimes associated with seizures. CASE SUMMARY: This report describes an 85-year-old Chinese male inpatient who twice received concomitant administration of meropenem and valproic acid for the treatment of pneumonia and poststroke epilepsy, respectively. Rapid declines in valproic acid concentrations were observed both times after meropenem administration. No seizures occurred in the first treatment period; however, when the patient suffered pneumonia again 3 months later, the same concomitant therapy was prescribed, and seizures occurred. It is difficult to identify a single etiology of the seizures. Based on a score of 7 on the Naranjo adverse drug reaction probability scale, the seizures were considered to be probably related to the concomitant administration of meropenem and valproic acid. CONCLUSIONS: Various factors make the effect of concomitant administration of meropenem and valproic acid unpredictable, even in the same patient. Caution should be used when administering meropenem and valproic acid concomitantly, especially in elderly patients with central nervous system disorders, even if the patient has had a successful prior experience with these 2 drugs. If concomitant administration is essential, very close serum concentration monitoring and clinical observation are necessary. PMID- 19281942 TI - Darwin's theory revisited by today's biology. PMID- 19281943 TI - The evolution of creationists in the United States: where are they now, and where are they going? AB - The history of anti-evolutionism in the United States begins only in the early decades of the 20th century but has evolved considerably since then. Various versions of the movement ("equal time" for creationism, "creation science", "intelligent design") have developed over time, but they have made few positive contributions to serious discourse about science and religion. Their main goal has been to try to stop the teaching of evolution. The most recent version of creationism, "intelligent design" (ID), has little in common with William Paley's 18th-century version: ID posits an interventionist Deity who regularly interferes in natural processes to produce complex biological structures and functions. The 2005 "intelligent design" trial in Dover, Pennsylvania, destroyed any pretensions that the movement had to scientific integrity. However, anti-evolutionists continue to foment discord at local levels, where opposition to the teaching of evolution can be presented without strong resistance. Scientists can best demonstrate their concern by becoming involved in federal, state, and local administrative processes that determine curricula and develop and adopt textbooks and other instructional materials. PMID- 19281939 TI - Methylphenidate for the treatment of depressive symptoms, including fatigue and apathy, in medically ill older adults and terminally ill adults. AB - BACKGROUND: Depressive symptoms, fatigue, and apathy are common symptoms among medically ill older adults and patients with advanced disease, and have been associated with morbidity and mortality. Methylphenidate has been used to treat these symptoms because of its rapid effect. Despite the long history of methylphenidate use for the treatment of depressive symptoms, fatigue, and apathy, there is little definitive evidence to support its use. OBJECTIVE: The aim of this paper was to review the efficacy and tolerability of methylphenidate in the treatment of depressive symptoms, fatigue, and apathy in medically ill older adults and adults receiving palliative care. METHODS: English-language articles presenting systematic reviews, clinical trials, or case series describing the use of methylphenidate for the treatment of depressive symptoms, fatigue, or apathy in medically ill older adults or adults receiving palliative care were identified. The key words methylphenidate and either depressive, depression, fatigue, or apathy were used to search the Cochrane Database, MEDLINE, PsycINFO, and International Pharmaceutical Abstracts. Included articles addressed depressive symptoms, fatigue, or apathy in (1) older adults (generally, age > or =65 years), particularly those with comorbid medical illness; (2) adults receiving palliative care; and (3) adults with other chronic illnesses. I excluded articles regarding treatment of depression in healthy young adults; bipolar disorder and attention-deficit/hyperactivity disorder; and narcolepsy, chronic fatigue syndrome, and related disorders. RESULTS: A total of 19 controlled trials of methylphenidate in medically ill older adults or patients in palliative care were identified. Unfortunately, their conflicting results, small sample sizes, and poor methodologic quality limited the ability to draw inferences regarding the efficacy of methylphenidate, although evidence of tolerability was stronger. The available evidence suggests possible effectiveness of methylphenidate for depressive symptoms, fatigue, and apathy in various medically ill populations. CONCLUSION: In the absence of definitive evidence of effectiveness, trials of low-dose methylphenidate in medically ill adults with depression, fatigue, or apathy, with monitoring for response and adverse effects, are appropriate. PMID- 19281944 TI - [Haeckel: a German Darwinian?]. AB - German biologist Ernst Haeckel (1834-1919) is often considered the most renowned Darwinian in his country since, as early as 1862, he declared that he accepted the conclusions Darwin had reached three years before in On the Origin of Species, and afterwards, he continuously proclaimed himself a supporter of the English naturalist and championed the evolutionary theory. Nevertheless, if we examine carefully his books, in particular his General Morphology (1866), we can see that he carries on a tradition very far from Darwin's thoughts. In spite of his acceptance of the idea of natural selection, that he establishes as an argument for materialism, he adopts, indeed, a conception of evolution that is, in some respects, rather close to Lamarck's views. He is, thus, a good example of the ambiguities of the reception of Darwinism in Germany in the second part of the 19th century. PMID- 19281945 TI - Epiphytism in ferns: diversity and history. AB - As for other vascular plants, numerous adaptive strategies have been selected in epiphytic ferns in order to survive in a constraining and desiccating environment and thus to prevent dehydration and/or to access to water and nutrients. Here we present some of the specializations that allow ferns to survive in this particular habitat. Some of the most spectacular epiphytic specializations are observed in the Polypodiaceae family, involving humus-collectors which entrap humus in specialized organs, and ant-plant mutualism strategies. We then address the question of epiphytism in an evolutionary context. There is little fossil evidence of vascular epiphytes. Inferring the evolution of epiphytism in extant ferns shows that diversification of major living epiphytic groups mostly occurred in the Tertiary. Finally, we focus on the Hymenophyllaceae family which provides an original example of hygrophilous epiphytic strategy that is unique in vascular plants. PMID- 19281946 TI - The molecular evolutionary tree of lizards, snakes, and amphisbaenians. AB - Squamate reptiles (lizards, snakes, amphisbaenians) number approximately 8200 living species and are a major component of the world's terrestrial vertebrate diversity. Recent molecular phylogenies based on protein-coding nuclear genes have challenged the classical, morphology-based concept of squamate relationships, requiring new classifications, and drawing new evolutionary and biogeographic hypotheses. Even the key and long-held concept of a dichotomy between iguanians (approximately 1470 sp.) and scleroglossans (all other squamates) has been refuted because molecular trees place iguanians in a highly nested position. Together with snakes and anguimorphs, iguanians form a clade- Toxicofera--characterized by the presence of toxin secreting oral glands and demonstrating a single early origin of venom in squamates. Consequently, neither the varanid lizards nor burrowing lineages such as amphisbaenians or dibamid lizards are the closest relative of snakes. The squamate timetree shows that most major groups diversified in the Jurassic and Cretaceous, 200-66 million years (Myr) ago. In contrast, five of the six families of amphisbaenians arose during the early Cenozoic, approximately 60-40 Myr ago, and oceanic dispersal on floating islands apparently played a significant role in their distribution on both sides of the Atlantic Ocean. Among snakes, molecular data support the basic division between the small fossorial scolecophidians (approximately 370 sp.) and the alethinophidians (all other snakes, approximately 2700 sp.). They show that the alethinophidians were primitively macrostomatan and that this condition was secondarily lost by burrowing lineages. The diversification of alethinophidians resulted from a mid-Cretaceous vicariant event, the separation of South America from Africa, giving rise to Amerophidia (aniliids and tropidophiids) and Afrophidia (all other alethinophidians). Finally, molecular phylogenies have made it possible to draw a detailed evolutionary history of venom among advanced snakes (Caenophidia), a key functional innovation underlying their radiation (approximately 2500 sp.). PMID- 19281947 TI - Progress in systematics: from Siboglinidae to Pogonophora and Vestimentifera and back to Siboglinidae. AB - We review the taxonomic history of pogonophores (frenulates and vestimentiferans), from the species in first described 1914 to the recently described bone-eating worm Osedax. Previous systematists have referred both groups to the rank of phylum, and the animals have been treated as deuterostomes with a dorsal nerve cord. Further knowledge on their embryology, the discovery of the previously overlooked posterior, segmented part provided with chaetae, and access to molecular data, have completely changed earlier views on their affinities. They are now referred to as a single family of polychaete annelids, Siboglinidae. PMID- 19281948 TI - Lessons from parasitic flatworms about evolution and historical biogeography of their vertebrate hosts. AB - Cophylogenetic studies investigate the evolutionary trends within host-parasite associations. Examination of the different levels of fidelity between host and parasite phylogenies provides a powerful tool to inspect patterns and processes of parasite diversification over host evolution and geological times. Within the phylum Platyhelminthes, the monogeneans are mainly fish parasites. The Polystomatidae, however, are known from the sarcopterygian Australian lungfish and tetrapods such as amphibians, freshwater turtles, and the African hippopotamus. Cophylogenetic and biogeographic vicariance analyses, supplemented by molecular calibrations, showed that the Polystomatidae may track the evolutionary history of the first aquatic tetrapods in the Palaeozoic age. Evolutionary lines of the major polystome lineages would also be intimately related to the evolution of their hosts over hundreds of millions years. Since the Mesozoic, evolution of polystomes would have been shaped mainly by plate tectonics during the break-up of Gondwanaland and subsequent dispersal of ancestral neobatrachian host lineages. Therefore the Polystomatidae could serve as a novel model to improve cophylogenetic tools and to inspect a suite of questions about the evolution of vertebrate hosts. PMID- 19281949 TI - Diversity and evolution of marine phytoplankton. AB - Marine phytoplankton organisms account for more than 45% of the photosynthetic net primary production on Earth. They are distributed across many of the major clades of the tree of life and include prokaryotes, and eukaryotes that acquired photosynthesis through the process of endosymbiosis. If the number of extant described species is relatively low compared to the diversity of the terrestrial plants, recent insights into the genetic diversity of natural assemblages have revealed a large unsuspected diversity at different taxonomic levels. Wide infra specific diversity is also being discovered in many widespread and well known morphological species. This review summarizes data obtained in the fields of ecology, evolutionary biology, physiology and genomics that have improved our understanding of the biodiversity and evolution of marine phytoplankton. PMID- 19281950 TI - Molecular phylogeny: reconstructing the forest. AB - Phylogeny, be it morphological or molecular, has long tried to explain the extant biodiversity by the Tree of Species, which is a logical consequence of strict Darwinian evolutionary principles. Through constant improvement of both methods and data sets, some parts of this diversity have actually been demonstrated to be the result of a tree-like process. For some other parts, and especially for prokaryotes, different molecular markers have, however, produced different evolutionary trees, preventing the reconstruction of such a Tree. While technical artifacts could be blamed for these discrepancies, Lateral Gene Transfers are now largely held for responsible, and their existence requires an extension of the Darwinian framework, since genetic material is not always vertically inherited from parents to offspring. Through a variety of biological processes, sometimes large parts of DNA are exchanged between phylogenetically distant contemporary organisms, especially between those sharing the same environment. While mainly concerning prokaryotes, Lateral Gene Transfers have been also demonstrated to affect eukaryotes, and even multicellular ones, like plants or animals. Most of the time, these transfers allow important adaptations and the colonisation of new niches. The quantitative and qualitative importance of genetic transfers has thus severely challenged the very existence of a universal Tree of Species, since genetic connections, at least for microbes, seem more reticulated than tree-like. Even traditional biological concepts, like the concept of species, need to be re evaluated in the light of recent discoveries. In short, instead of focusing on a elusive universal tree, biologists are now considering the whole forest corresponding to the multiple processes of inheritance, both vertical and horizontal. This constitutes the major challenge of evolutionary biology for the years to come. PMID- 19281951 TI - Early evolution of symmetry and polarity in metazoan body plans. AB - The early diverging metazoan lineages have highly disparate adult body plan geometries, which can be characterised in terms of five major types of symmetry (asymmetrical, spherical, cylindrical, n-radial, bilateral). Patterns of evolutionary changes in symmetry types and the homology of body axes across lineages are discussed here by confronting evidence from comparative anatomy, phylogeny, genomics and evo-devo. The conventional scenario, postulating a graded complexification from asymmetry to radial and finally bilateral symmetry, is considered untenable. Cylindrical symmetry is likely to be the ancestral type from which derived all remaining types through multiple convergences. Recent proposals prompted by molecular data that the bilateral anatomies of many cnidarians and of the Bilateria are homologous are clearly not supported. The Hox based patterning system operating along the antero-posterior axis of the Bilateria does not seem to predate their divergence with the Cnidaria, but intercellular signalling systems, notably the Wnt pathway, could have been involved in generating the main body axis in the last common ancestor of the Metazoa. PMID- 19281952 TI - Towards a synthetic view of axis specification mechanisms in vertebrates: insights from the dogfish. AB - The genetic mechanisms, which control axis specification, apparently extensively diverge across vertebrates. In amphibians and teleosts, they are tightly linked to the establishment of an early dorso-ventral polarity. This polarity has no equivalent in amniotes, which unlike the former, retain a considerable plasticity for their site of axis formation until blastula stages and rely on signals secreted by extra-embryonic tissues for the establishment of their early rostro caudal pattern. In order to better understand the links between these seemingly highly divergent mechanisms, we have used an evo-devo approach, aimed at reconstructing the gnathostome ancestral state and focussed on a chondrichthyan, the dogfish Scyliorhinus canicula. A detailed molecular characterization of the dogfish embryo at blastula and gastrula stages highlights striking similarities with all vertebrate model organisms including amniotes. It suggests the presence in the dogfish of territories homologous to the hypoblast and extra-embryonic ectoderm of the latter, which may therefore reflect the primitive condition of jawed vertebrates. In the ancestral state, these territories are specified at opposite sides of an early axis of bilateral symmetry, homologous to the dorso ventral axis of amphibians and teleosts, and aligned with the later forming embryonic axis, from head to tail. Amniotes have diverged from this pattern through a posterior expansion of extra-embryonic ectoderm, resulting in an apparently radial symmetry at late blastula stages. These data delineate the broad outlines of the gnathostome ancestral pattern of axis specification and highlight an unexpected unity of mechanisms across jawed vertebrates. They illustrate the complementarity of comparative and genetic approaches for a comprehensive view of developmental mechanisms themselves. PMID- 19281953 TI - Evolution of floral symmetry: a state of the art. AB - The genetic determinants of the organisation and variation of the flower, a striking feature of the angiosperms, are only beginning to be deciphered. Floral symmetry has recurrently evolved among angiosperms, zygomorphy (monosymmetry) being a key innovation due to its role in the plant-pollination interaction. As such, it represents a case study for evo-devo. Phylogenetic comparative studies conducted in two eudicot clades, the Ranunculales and the Asteridae sensu APGII, have shown that the evolution of this trait is dependent upon the architectural context of the flower. Genetic and developmental bases of zygomorphy have been investigated in several unrelated model species. In all these species, zygomorphy appears to be controlled, at least partially, by genes belonging to the TCP gene family of transcription factors and named CYC-like genes. Exploring the molecular bases of zygomorphy in non-model species spanning the diversity of angiosperms, but also the developmental processes involved, are now essential to understand the evolution of floral symmetry. PMID- 19281954 TI - Fluidity of eukaryotic genomes. AB - The understanding the different kinds of sequences that make up a genome, as well as their proportions in genomes (obtained by the sequencing of the complete genome), has considerably changed our idea of evolution at the genomic level. The former view of a slowly evolving genome has given way to the idea of a genome that can undergo many transformations, on a large or small scale, depending on the evolution of the different types of sequences constituting it. Here we summarise the evolution of these sequences and the impact it can have on the genome. We have focused on micro-transformations, and especially on the impact of transposable elements on genomes. PMID- 19281955 TI - "Changing by doubling", the impact of Whole Genome Duplications in the evolution of eukaryotes. AB - Species are usually defined by reproductive isolation and are characterized by their gene repertoire. These two aspects are consequences of events fixed during evolution, including whole genome duplications and other polyploidizations. Thanks to the recent progress in genome sequencing, new light has been shed on these events. In this review, we will summarize these findings and discuss the methodology involved. Evolutionary traces of such events have been evidenced in various lineages in plants, animals, fungi and protozoa. Comparative analysis of synteny is a powerful approach to unveil evolutionary footprints of these events. According to expectations, these events would facilitate speciation since some of them are thought to be at the base of major radiations such as teleostei or eudicotyledons. After an initial amplification, the gene repertoire would be shaped by constraints such as expression level and functional interactions that would tend to maintain only a tiny fraction of the duplicates over the long term. Functional innovation from duplication may be a secondary effect, enabled by these duplicate retention mechanisms. PMID- 19281956 TI - A prominent role for segmental duplications in modeling eukaryotic genomes. AB - Segmental duplications (SDs) are a major element of eukaryotic genomes. Whereas their quantitative importance vary among lineages, SDs appear as a fundamental trait of the recent evolution of great-apes genomes. The chromosomal instability generated by these SDs has dramatic consequences both in generating a high level of polymorphisms among individuals and in originating numerous human pathogenic diseases. However, even though the importance of SDs has been increasingly recognized at the genomic level, some of the molecular pathways that lead to their formation remain obscure. Here we review recent evidences that the interplay between several mechanisms, some conservative, some based on replication, explains the complex SDs patterns observed in many genomes. Recent experimental studies have indeed partially unveiled some important aspects of these mechanisms, shedding interesting and unsuspected new lights on the dramatic plasticity of eukaryotic genomes. PMID- 19281957 TI - The molecular bases of cereal domestication and the history of rice. AB - In this review, we discuss the development of molecular genetics and genomics that has allowed one to identify and characterize some of the key genes involved in cereal domestication. The list is far from being complete, but the first conclusion that can be drawn from the published works is that only a few loci have been the target of human selection in the first stages of the domestication process at the late neolithic. Mutations at these few loci have led to dramatic changes in plant morphology and phenology, transforming a wild into a cultivated plant. We also show that in the case of rice, for which the complete genome sequence is available, the development of new molecular markers based on retrotransposon insertion polymorphisms helped to resolve some of the questions regarding the origin of the domestication of the crop in Asia. PMID- 19281958 TI - Structure and function evolution in the superfamily of globins. AB - The superfamily of globins has emerged some 4000 Myr from a common ancestor, which was among the basic protein components required for life. Globins are present in the three kingdoms of life. From a structure point of view, these molecules are defined by the presence of a characteristic protein fold, rich in alpha-helix, surrounding a heme group. Depending on the species or organs, they may be physiologically active as monomers, tetramers or large size polymers. Their function varies from the classical reversible binding of oxygen for transport and storage to cytoprotection against reactive oxygen species, NO scavenging, signaling in oxygen dependent metabolic pathways, or possibly other specific properties involving ligand or electron transfer. All these aspects are discussed in this review. PMID- 19281959 TI - Infection by Wolbachia: from passengers to residents. AB - Wolbachia are endosymbiotic alpha-proteobacteria harboured by terrestrial arthropods and filarial nematodes, where they are maternally transmitted through egg cytoplasm. According to the host group, Wolbachia have developed two contrasting symbiotic strategies. In arthropods, symbiosis is secondary (i.e. facultative), and Wolbachia insure their transmission as reproduction parasites. However, despite of the efficiency of the manipulation mechanisms used, Wolbachia are limited to the state of passenger because some factors can prevent the association between Wolbachia and their hosts to become permanent. On the contrary, symbiosis is primary (i.e. obligatory) in filarial nematodes where Wolbachia insure their transmission via a mutualistic relationship, leading them to become permanent residents of their hosts. However, a few examples show that in arthropods too some Wolbachia have started to present the first stages of a mutualistic behaviour, or are even truly indispensable to their host. Whatever its strategy, Wolbachia infection is a spectacular evolutionary success, this symbiotic bacterium representing one of the most important biomass of its kind. PMID- 19281960 TI - Symbioses between deep-sea mussels (Mytilidae: Bathymodiolinae) and chemosynthetic bacteria: diversity, function and evolution. AB - Mussels of the subfamily Bathymodiolinae thrive around chimneys emitting hot fluids at deep sea hydrothermal vents, as well as at cold seeps and on sunken organic debris (sunken wood, whale falls). Despite the absence of light-driven primary production in these deep-sea ecosystems, mussels succeed reaching high biomasses in these harsh conditions thanks to chemosynthetic, carbon-fixing bacterial symbionts located in their gill tissue. Since the discovery of mussel symbioses about three decades ago our knowledge has increased, yet new findings are published regularly regarding their diversity, role and evolution. This article attempts to summarize current knowledge about symbiosis in Bathymodiolinae, focusing on mussel species for which information is available regarding both hosts and symbionts. Moreover, new data obtained from small mussels inhabiting sunken woods around the Philippines are provided. Indeed, mussel species from organic falls remain poorly studied compared to their vent and seep relatives despite their importance for the understanding of the evolution of symbiosis in the subfamily Bathymodiolinae. PMID- 19281961 TI - Virulence strategies in parasitoid Hymenoptera as an example of adaptive diversity. AB - Parasitoids are mostly insects that develop at the expense of other arthropods, which will die as a result of the interaction. Their reproductive success thus totally depends on their ability to successfully infest their host whose reproductive success relies on its own ability to avoid or overcome parasitism. Such intense selective pressures have resulted in extremely diverse adaptations in parasitoid strategies that ensure parasitism success. For instance, wasp specific viruses (polydnaviruses) are injected into the host by parasitoid females to modulate its physiology and immunity. This article synthesizes available physiological and molecular data on parasitoid virulence strategies and discusses the evolutionary processes at work. PMID- 19281962 TI - When domestic cat (Felis silvestris catus) population structures interact with their viruses. AB - Many theoretical studies have proposed different causal mechanisms by which the structure of a host population could have important implications for life history traits of pathogens. However, little information is available from real systems to test these hypotheses. The domestic cat, Felis silvestris catus, whose populations exhibit a great variability in social and spatial structure, represent an ideal case study to assess this question. In the present article, we show how cat population structure may have influenced the evolution of feline viruses and, in return, how these viruses may have modified the genetic structure of cat populations. PMID- 19281963 TI - Effects of instantaneous object input and past experience on object-based attention. AB - Evidence for object-based attention typically comes from studies using displays with unchanged objects, and no consensus has yet been reached as to whether the object effect would be altered by changing object displays or having seen this change across-trials. We examined this by using modifications of the double rectangle cuing paradigm of Egly et al. [Egly, R., Driver, J., & Rafal, R. D. (1994). Shifting visual attention between objects and locations: Evidence from normal and parietal lesion subjects. Journal of Experimental Psychology: General, 123, 161-177], and our results, when the objects remained unchanging, replicated the original object effect. However, no object effect was found when the rectangles disappeared from view in the last (target) frame. This was true regardless of the likelihood of the rectangles disappearing, indicating the importance of instantaneous object inputs for object-based attention. The across trial experience of seeing a different object (boomerang), however, was found to influence the object effect when the cued rectangles persisted throughout the trial. Unlike previous studies, which emphasize one or the other, we demonstrate clearly that instantaneous object inputs and past experience interact to determine the way attention selects objects. PMID- 19281964 TI - [Brain-Derived Neurotrophic Factor (BDNF): role of this neurotrophin in cardiovascular physiopathology]. AB - Neurotrophic-factors research is dominated by neurotrophins (NT): a family of polypeptides which includes molecules such as Nerve Growth Factor (NGF) and the Brain-Derived Neurotrophic Factor (BDNF). They are homodimeric polypeptides. NTs interact with classes of receptors on responsive cells: protein-tyrosine kinase type receptors (Trk family). It is well established that the levels of NT determine the balance between cell survival and apoptosis during neural development. Recently, it has been shown that BDNF played a role in the etiology of some cardiovascular diseases: induction of angiogenesis in ischemic issues. Plasma BDNF was increased in the circulation in patients with unstable angina. BDNF was expressed in atheromatous intima and adventitia in human coronary artery. Our own studies suggest that BDNF serum levels in patients with acute myocardial infarction or under cardiopulmonary bypass could related to platelet activation, oxidative stress and inflammatory response. Thus, investigations of this new factor: BDNF will help to better understand vascular development and may lead to new therapeutic strategies for some cardiovascular diseases. PMID- 19281965 TI - Use of empirically supported interventions for psychopathology: can the participatory approach move us beyond the research-to-practice gap? AB - Dissemination, or distribution, of empirically supported interventions (ESIs) for psychopathology remains a significant challenge. This paper reviews the principles of community-partnership research (CPR) and explores why CPR might improve distribution of psychological ESIs. Benefits of CPR include building trust, pooling resources and knowledge, and better serving a community by directly involving its members in the design and implementation of research. In addition, after establishing a community's trust using CPR, researchers are likely to be better positioned to partner with communities in the further distribution of ESIs via community networks. This paper reviews the case of dissonance-based eating disorder prevention interventions to provide an example of how CPR can facilitate the adoption and distribution of an ESI by a community, in this case, sororities. CPR also presents a number of challenges, however, because it is time consuming and does not always align with funding mechanisms and research designs used in randomized controlled trials. Further, CPR does not necessarily solve the challenge of training providers, though it may help with problem solving. Ultimately, we suggest that the benefits of CPR far outweigh the challenges, and hope that more researchers will adopt these practices so that more individuals can benefit from empirically supported psychological interventions. PMID- 19281966 TI - How to handle anxiety: The effects of reappraisal, acceptance, and suppression strategies on anxious arousal. AB - It has been suggested that reappraisal strategies are more effective than suppression strategies for regulating emotions. Recently, proponents of the acceptance-based behavior therapy movement have further emphasized the importance of acceptance-based emotion regulation techniques. In order to directly compare these different emotion regulation strategies, 202 volunteers were asked to give an impromptu speech in front of a video camera. Participants were randomly assigned to one of three groups. The Reappraisal group was instructed to regulate their anxious arousal by reappraising the situation; the Suppression group was asked to suppress their anxious behaviors; and the Acceptance group was instructed to accept their anxiety. As expected, the Suppression group showed a greater increase in heart rate from baseline than the Reappraisal and Acceptance groups. Moreover, the Suppression group reported more anxiety than the Reappraisal group. However, the Acceptance and Suppression groups did not differ in their subjective anxiety response. These results suggest that both reappraising and accepting anxiety is more effective for moderating the physiological arousal than suppressing anxiety. However, reappraising is more effective for moderating the subjective feeling of anxiety than attempts to suppress or accept it. PMID- 19281967 TI - Preparative production of colominic acid oligomers via a facile microwave hydrolysis. AB - The hydrolysis of colominic acid via microwave irradiation was studied for the production of short-chain oligomers with a degree of polymerization (DP) of 1-5. This method was compared to the traditional acid hydrolytic method for the production of preparative quantities of short colominic acid oligomers. The oligomers were purified by size exclusion chromatography and characterized by (1)H NMR. Optimal conditions for producing the dimer were found to be 12 min at 10% power in a 1000-Watt domestic microwave. This method is advantageous over the traditional technique in that the hydrolysis can be completed in just a few minutes, rather than in a few hours, it is reproducible, and yields large quantities of the desirable short chain oligomers of colominic acid. PMID- 19281968 TI - DFT study of the interaction of cytidine and 2'-deoxycytidine with Li+, Na+, and K+: effects of metal cationization on sugar puckering and stability of the N glycosidic bond. AB - Density functional theory (DFT) calculations were performed at the B3LYP level with a 6-311++G(d,p) basis set to systematically explore the geometrical multiplicity and binding strength for complexes formed by Li(+), Na(+), and K(+) with cytidine and 2'-deoxycytidine. All computational studies indicate that the metal ion affinity (MIA) decreases from Li(+) to Na(+) and K(+) for cytosine nucleosides. For example, for cytidine the affinity for the above metal ions are 79.5, 55.2, and 41.8 and for 2'-deoxycytidine, 82.8, 57.4, and 42.2 kcal/mol, respectively. It is also interesting to mention that linear correlations between calculated MIA values and the atomic numbers (Z) of the above metal ions were found. The influence of metal cationization on the coordination modes and the strength of the N-glycosidic bond in cytosine nucleosides have been studied. In all cases, the N1-C1' bond distance changes upon introducing a positive charge in the nucleosides. It has been found that metal binding significantly changes the values of the phase angle of pseudorotation P in the sugar unit of these nucleosides. With respect to the sugar ring, metal binding changes the values of the glycosyl torsion angle and sugar ring conformation. The present calculations in the gas phase provide the first clues on the intrinsic chemistry of these systems and may be of value for studies of the influence of metal cations on the conformational behavior and function of nucleic acids. PMID- 19281969 TI - Synthesis of the starfish ganglioside LLG-3 tetrasaccharide. AB - The first synthesis of the ganglioside LLG-3 tetrasaccharide, which has attractive biological activities as well as a unique structure, is described. A C8-methoxy decorated sialic acid building block was initially prepared and a glycolic acid moiety was then introduced by sialylation. Amide condensation between the sialyl glycolic acid and an amino group at C5 on the sialyllactoside unit afforded the fully protected LLG-3 tetrasaccharide. Finally, the desired tetrasaccharide part of LLG-3 was obtained after careful global deprotection. [structure: see text]. PMID- 19281970 TI - [Resection of a giant hepatic lymph node]. PMID- 19281971 TI - Novel-view scene recognition relies on identifying spatial reference directions. AB - Five experiments investigated whether observer locomotion provides specialized information facilitating novel-view scene recognition. Participants detected a position change after briefly viewing a desktop scene when the table stayed stationary or was rotated and when the observer stayed stationary or locomoted. The results showed that 49 degrees novel-view scene recognition was more accurate when the novel view was caused by observer locomotion than when the novel view was caused by table rotation. However such superiority of observer locomotion disappeared when the to-be-tested viewpoint was indicated during the study phase, when the study viewing direction was indicated during the test phase, and when the novel test view was 98 degrees , and was even reversed when the study viewing direction was indicated during the test phase in the table rotation condition but not in the observer locomotion condition. These results suggest scene recognition relies on the identification of the spatial reference directions of the scene and accurately indicating the spatial reference direction can facilitate scene recognition. The facilitative effect of locomotion occurs because the spatial reference direction of the scene is tracked during locomotion and more accurately identified at test. PMID- 19281972 TI - On the optimality of serial and parallel processing in the psychological refractory period paradigm: effects of the distribution of stimulus onset asynchronies. AB - Within the context of the psychological refractory period (PRP) paradigm, we developed a general theoretical framework for deciding when it is more efficient to process two tasks in serial and when it is more efficient to process them in parallel. This analysis suggests that a serial mode is more efficient than a parallel mode under a wide variety of conditions and thereby suggests that ubiquitous evidence of serial processing in PRP tasks could result from performance optimization rather than from a structural bottleneck. The analysis further suggests that the experimenter-selected distribution of stimulus onset asynchronies (SOAs) influences the relative efficiency of the serial and parallel modes, with a preponderance of short SOAs favoring a parallel mode. Experiments varying the distribution of SOAs were conducted, and the results suggest that there is a shift from a more serial mode to a more parallel mode as the likelihood of short SOAs increases. PMID- 19281973 TI - An efficient sliding window strategy for accurate location of eukaryotic protein coding regions. AB - The sliding window is one of important factors that seriously affect the accuracy of coding region prediction and location for the methods based on power spectrum technique. It is very difficult to select the appropriate sliding step and the window length for different organisms. In this study, a novel sliding window strategy is proposed on the basis of power spectrum analysis for the accurate location of eukaryotic protein coding regions. The proposed sliding window strategy is very simple and the sliding step of window is changeable. Our tests show that the average location error for the novel method is 12 bases. Compared with the previous location error of 54 bases using the fixed sliding step, the novel sliding window strategy increased the location accuracy greatly. Further, the consumed CPU time to run the novel strategy is much shorter than the strategy of the fixed length sliding step. So, the computational complexity for the novel method is decreased greatly. PMID- 19281974 TI - On the use of pseudo-spectral method in model reduction and simulation of active dendrites. AB - Most of dendrites in the central nervous system are now known to have active channels. These active dendrites play important roles not only in signal summation but also in computation. For the simulation of these important active dendrites, the compartment model based on the finite volume or finite difference discretization was mainly adopted. In this paper, we employ the Chebychev pseudo spectral method well developed in computational physics, and demonstrate that it can achieve a higher precision with the same number of equations than the compartment model. Moreover, it is also shown that the Chebychev pseudo-spectral method converges faster to attain a given precision. Hence, for the simulations of active dendrites, the Chebychev pseudo-spectral method can be an attractive alternative to the compartment model since it leads to a low order model with higher precision or converges faster for a given precision. PMID- 19281975 TI - Asthma in humans and cats: is there a common sensitivity to aeroallegens in shared environments? AB - Cats spontaneously develop eosinophilic airway inflammation and airway hyperreactivity that is very similar to human allergic asthma. In addition, household cats share environmental exposures to aeroallergens with humans. We review the scientific literature concerning the pathophysiology of feline asthma, including similarities to human asthma and evidence regarding environmental aeroallergen triggers. Results of pathophysiological studies suggest important similarities between human and feline responses to inhaled allergens. Only a few studies were found that examined the development of disease in cats to environmental aeroallergens. Limited evidence suggests that some environmental allergens can cause disease in both cats and humans. It appears that there is a need for greater communication between human and animal health professionals regarding environmental causes of asthma. Specifically, additional research into linkages between human and feline asthma using both molecular techniques and clinical epidemiological approaches could lead to improved understanding of the environmental risks. Finally, there should be consideration of use of naturally affected and/or experimentally induced (using clinically relevant allergens) asthmatic cats in preclinical trials for novel therapeutic interventions. PMID- 19281976 TI - Re: Minimally invasive ilioinguinal approach to the acetabulum. PMID- 19281977 TI - Finite element analysis, mechanical assessment and material comparison of two volar slab constructs. AB - INTRODUCTION: In order to help prevent joint stiffness, the injured or postoperative hand is splinted in the intrinsic-plus position. We aimed to determine the strongest type of volar slab construct that would be appropriate in this treatment. METHODS: Two constructs were compared, a double-ridged and a non ridged slab. Two materials were used, plaster of Paris (POP) and resin. We performed finite element analysis (FEA) and mechanical assessment to establish which combination of construct and material resulted in the strongest volar slab. We were unable to form ridges on the resin slab, and therefore this construct was not tested. RESULTS: Finite element analysis showed that most stress occurred at the wrist region, where all the slabs failed. The double-ridged POP slab was found to be 5.3 times stronger than the non-ridged POP slab and 1.4 times stronger than the non-ridged resin slab (p<0.001). CONCLUSION: To improve strength of the conventional POP volar slab, we suggest forming two anterior ridges in the plaster. PMID- 19281978 TI - Early diagnosis of tibial compartment syndrome: continuous pressure measurement or not? PMID- 19281979 TI - Re: Development of a femoral non-union model in the mouse. PMID- 19281980 TI - Prevention of intra-operative contamination from surgical gown cuffs. PMID- 19281981 TI - The association between fatal vascular events and risk factors for carotid atherosclerosis in patients on maintenance hemodialysis: plaque number of dialytic atherosclerosis study. AB - Atherosclerotic vascular diseases are a major cause of morbidity and mortality for end-stage renal disease patients. We followed prospectively 226 hemodialysis patients by carotid ultrasonography to determine if ultrasonographic markers are predictive of the prognosis of these patients. The end-point was death or completion of the five-year follow-up period. Fatal cerebrovascular and cardiovascular events were the most common cause of death. By multivariate analysis, diabetes mellitus (DM) (P=0.005), plaque number (PN) by ultrasonography (P=0.023), age (P=0.001), calcium-phosphate product (P=0.049), and serum albumin (P=0.009) were extracted as independent risk factors. The five-year increase of PN was significantly greater for DM patients than for non-DM patients. Moreover, PN was an independent marker of a fatal event, irrespective of DM status. Our results suggest that PN may be a useful predictor of the long-term prognosis of hemodialysis patients. PMID- 19281982 TI - Insights into atherosclerosis from invasive and non-invasive imaging studies: Should we treat subclinical atherosclerosis? AB - Although atherosclerosis is associated with the elderly, young adults with hypercholesterolemia and other cardiovascular risk factors may have subclinical atherosclerotic disease. In many cases, when two or more risk factors are present, conventional risk assessment using the Framingham score, that was not designed to detect atherosclerotic plaques, may significantly underestimate the extent of atherosclerosis. Several non-invasive imaging technologies now make it possible to identify subclinical atherosclerosis before symptoms appear or major vascular events occur. These include B-mode ultrasound to measure carotid intima media thickness, computed tomography to measure coronary artery calcification, and high-resolution magnetic resonance imaging to evaluate plaque size and composition. On the basis of available evidence, assessment of subclinical atherosclerosis should be considered in persons judged to be at intermediate risk by Framingham score, because test results may influence risk stratification and, consequently, the intensity of therapeutic intervention. Patients with significant subclinical atherosclerosis are at high risk and, like other high risk individuals, should receive treatment designed to achieve aggressive low density lipoprotein cholesterol targets. Clinical studies show that statin therapy may delay atherosclerosis progression and that intensive therapy with rosuvastatin may actually reverse the atherosclerotic process. PMID- 19281984 TI - Subclinical atherosclerosis in association with elevated placental vascular resistance in early pregnancy. PMID- 19281985 TI - Osteopontin gene variation and cardio/cerebrovascular disease phenotypes. AB - We aimed at associating common osteopontin (OPN) gene variants with cardiovascular disease phenotypes.We scanned the OPN gene in 190 chromosomes from myocardial infarction (MI) patients and identified five variants in the promoter, three synonymous and one non-synonymous variant. All variants were investigated in case-control studies for MI (ECTIM: 990 cases, 900 controls) and brain infarction (BI) (GENIC: 466 cases, 444 controls). Promoter variants were functionally analyzed by bandshift assays, the coding D147D [T/C] by Western blot. Allele D147D C was independently and significantly associated with lower apoB levels (P=0.044 [ECTIM] P=0.03 [GENIC]), its allele frequency was significantly lower in patients with BI compared to controls (OR [95% CI] 0.39 [0.20-0.74], P=0.004), and C allele carriers had a significantly lower frequency of presence of carotid plaques (P=0.02). Bandshifts with HepG2 and Ea.hy926 nuclear proteins did not reveal any functionality of promoter variants, whereas the OPN-441C-containing construct resulted in reduced OPN protein expression in Western blots, complying with its potential protective effect on the phenotypes studied.We here provide evidence that a portion of the OPN locus is likely to associate with cardiovascular disease-related phenotypes. However, further experiments are warranted to clarify the functional role of OPN variants. PMID- 19281986 TI - Micro-inflammatory changes in asymptomatic healthy adults during bouts of respiratory tract infections in the community: potential triggers for atherothrombotic events. AB - OBJECTIVE: To explore the possibility that mild inflammatory changes exist in asymptomatic adults during bouts of acute respiratory tract infections in the general population, and may mediate atherothrombotic events. METHODS: An epidemiological study that enrolled 5315 males and 2795 females attending a routine screening health program between the years 2003 and 2007. We correlated the concentrations of high sensitivity C-reactive protein (hs-CRP) and quantitative fibrinogen in completely asymptomatic and non-inflamed adults to the weekly epidemiological data for the incidence of acute respiratory tract infections in the community. RESULTS: Significant seasonal variations in the inflammatory variables were found for both genders. The population's weekly rates of acute respiratory tract infection had a significant epidemiological influence on the inflammatory biomarkers in the asymptomatic cohort. The magnitude of this influence could reach as much as 12% (3-22%) in hs-CRP concentrations in women and 0.30 micromol/L (0.20-0.41) in fibrinogen concentrations in men, for the change between the mean August and the mean January population's respiratory illness burden. CONCLUSION: Increase in the concentrations of two inflammation sensitive biomarkers can occur in completely asymptomatic adults at times of increased burden of acute respiratory tract infection in the general population. The possibility exists that these inflammatory changes represent occult and asymptomatic infections that could by themselves trigger acute atherothrombotic events. PMID- 19281987 TI - Quantitative measures of femoral fracture repair in rats derived by micro computed tomography. AB - Although fracture healing is frequently studied in pre-clinical models of long bone fractures using rodents, there is a dearth of objective quantitative techniques to assess successful healing. Biomechanical testing is possibly the most quantitative and relevant to a successful clinical outcome, but it is a destructive technique providing little insight into the cellular mechanisms associated with healing. The advent of X-ray computed tomography (CT) has provided the opportunity to quantitatively and non-destructively assess bone structure and density, but it is unknown how measurements derived using this technology relate to successful healing. To examine possible relationships, we used a pre-clinical model to test for statistically significant correlations between quantitative characteristics of the callus by micro-CT (microCT) and the bending strength, stiffness, and energy-to-failure of the callus as assessed by three-point bending of excised bones. A closed, transverse fracture was generated in the mid-shaft of rat femurs by impact loading. Shortly thereafter, the rats received a one-time, local injection of either the vehicle or one of four doses of lovastatin. Following sacrifice after 4 weeks of healing, fractured femurs were extracted for microCT analysis and then three-point bending. Setting the region of interest to be 3.2 mm above and below the fracture line, we acquired standard and new microCT-derived measurements. The mineralized callus volume and the mineral density of the callus correlated positively with callus strength (rxy = -0.315, p = 0.016 and rxy = 0.444, p<0.0005, respectively) and stiffness (rxy = -0.271, p = 0.040 and rxy = 0.325, p = 0.013, respectively), but the fraction of the callus that mineralized and the moment of inertia of the callus did not. This fraction did correlate with energy-to-failure (rxy = -0.343, p = 0.0085). Of the microCT-derived measurements, quantifying defects within the outer bridging cortices of the callus produced the strongest correlation with both callus strength (rxy = 0.557, p<0.0001) and stiffness (rxy = 0.468, p = 0.0002). By both reducing structural defects and increasing mineralization, lovastatin appears to increase the callus strength. PMID- 19281988 TI - The role of the carotid sinus in the reduction of arterial wall stresses due to head movements--potential implications for cervical artery dissection. AB - Spontaneous dissection of the cervical internal carotid artery (sICAD) is a major cause of stroke in young adults. A tear in the inner part of the vessel wall triggers sICAD as it allows the blood to enter the wall and develop a transmural hematoma. The etiology of the tear is unknown but many patients with sICAD report an initiating trivial trauma. We thus hypothesised that the site of the tear might correspond with the location of maximal stress in the carotid wall. Carotid artery geometries segmented from magnetic resonance images of a healthy subject at different static head positions were used to define a path of motion and deformation of the right cervical internal carotid artery (ICA). Maximum head rotation to the left and rotation to the left combined with hyperextension of the neck were investigated using a structural finite element model. A role of the carotid sinus as a geometrically compliant feature accommodating extension of the artery is shown. At the extreme range of the movements, the geometrical compliance of the carotid sinus is limited and significant stress concentrations appear just distal to the sinus with peak stresses at the internal wall on the posterior side of the vessel following maximum head rotation and on the anteromedial portion of the vessel wall following rotation and hyperextension. Clinically, the location of sICAD initiation is 10-30 mm distal to the origin of the cervical ICA, which corresponds with the peak stress locations observed in the model, thus supporting trivial trauma from natural head movements as a possible initiating factor in sICAD. PMID- 19281989 TI - Reducing ground reaction forces in gymnastics' landings may increase internal loading. AB - The aim of this study was to use a subject-specific seven-link wobbling mass model of a gymnast, and a multi-layer model of a landing mat, to determine landing strategies that minimise ground reaction forces (GRF) and internal forces. Subject-specific strength parameters were determined that defined the maximum voluntary torque/angle/angular velocity relationship at each joint. These relationships were used to produce subject-specific 'lumped' linear muscle models for each joint. Muscle activation histories were optimised using a Simplex algorithm to minimise GRF or bone bending moments for forward and backward rotating vault landings. Optimising the landing strategy to minimise each of the GRF reduced the peak vertical and horizontal GRF by 9% for the backward rotating vault and by 8% and 48% for the forward rotating vault, compared to a matching simulation. However, most internal loading measures (bone bending moments, joint reaction forces and muscle forces) increased compared to the matching simulation. Optimising the landing strategy to minimise the peak bone bending moments resulted in reduced internal loading measures, and in most cases reduced GRF. Bone bending moments were reduced by 27% during the forward rotating vault and by 2% during the backward rotating vault landings when compared to the matching simulations. It is possible for a gymnast to modify their landing strategy in order to minimise internal forces and lower GRF. However, using a reduction in GRF, due to a change in landing strategy, as a basis for a reduction in injury potential in vaulting movements may not be appropriate since internal loading can increase. PMID- 19281990 TI - Letter to the Editor referring to the article 'Some basic relationship between density values in cancellous bone and cortical bone' published on Journal of Biomechanics (volume 41, Issue 9, Pages 1961-8). PMID- 19281991 TI - A frame-invariant formulation of Fung elasticity. AB - Fung elasticity refers to the hyperelasticity constitutive relation proposed by Fung and co-workers for describing the pseudo-elastic behavior of biological soft tissues undergoing finite deformation. A frame-invariant formulation of Fung elasticity is provided for material symmetries ranging from orthotropy to isotropy, which uses Lame-like material constants. In the orthotropic case, three orthonormal vectors are used to define mutually orthogonal planes of symmetry and associated texture tensors. The strain energy density is then formulated as an isotropic function of the Lagrangian strain and texture tensors. The cases of isotropy and transverse isotropy are derived from the orthotropic case. Formulations are provided for both material and spatial frames. These formulations are suitable for implementation into finite element codes. It is also shown that the strain energy function can be naturally uncoupled into a dilatational and a distortional part, to facilitate the computational implementation of incompressibility. PMID- 19281992 TI - In situ estimation of tendon material properties: differences between muscles of the feline hindlimb. AB - Recent experiments to characterize the short-range stiffness (SRS)-force relationship in several cat hindlimb muscles suggested that the there are differences in the tendon elastic moduli across muscles [Cui, L., Perreault, E.J., Maas, H., Sandercock, T.G., 2008. Modeling short-range stiffness of feline lower hindlimb muscles. J. Biomech. 41 (9), 1945-1952.]. Those conclusions were inferred from whole muscle experiments and a computational model of SRS. The present study sought to directly measure tendon elasticity, the material property most relevant to SRS, during physiological loading to confirm the previous modeling results. Measurements were made from the medial gastrocnemius (MG), tibialis anterior (TA) and extensor digitorum longus (EDL) muscles during loading. For the latter, the model indicated a substantially different elastic modulus than for MG and TA. For each muscle, the stress-strain relationship of the external tendon was measured in situ during the loading phase of isometric contractions conducted at optimum length. Young's moduli were assessed at equal strain levels (1%, 2% and 3%), as well as at peak strain. The stress-strain relationship was significantly different between EDL and MG/TA, but not between MG and TA. EDL had a more apparent toe region (i.e., lower Young's modulus at 1% strain), followed by a more rapid increase in the slope of the stress-strain curve (i.e., higher Young's modulus at 2% and 3% strain). Young's modulus at peak strain also was significantly higher in EDL compared to MG/TA, whereas no significant difference was found between MG and TA. These results indicate that during natural loading, tendon Young's moduli can vary considerably across muscles. This creates challenges to estimating muscle behavior in biomechanical models for which direct measures of tendon properties are not available. PMID- 19281993 TI - Preparative separation of di- and trisulfonated components of Quinoline Yellow using affinity-ligand pH-zone-refining counter-current chromatography. AB - Four positionally isomeric 2-(2-quinolinyl)-1H-indene-1,3(2H)-dionedisulfonic acids (SA) and one triSA, components of the color additive Quinoline Yellow (QY, Color Index No. 47005), were isolated from the dye mixture by affinity-ligand pH zone-refining counter-current chromatography (CCC) through complementary use of ion-exchange and ion-pair reagents as the ligand. The added ligands facilitated the partitioning of the very polar polysulfonated components into the organic stationary phase of the two-phase solvent systems that consisted of isoamyl alcohol-methyl tert-butyl ether-acetonitrile-water (3:5:1:7), (3:4:1:7) or (3:1:1:5). Thus, separation of a 5-g portion of QY using sulfuric acid as the retainer and dodecylamine as the ligand (an ion-exchange reagent, 20% in the stationary phase), resulted in 1.21g of 6',5-diSA and 1.69g of 6',8',5-triSA, both of over 99% purity. A minor component, 8',4-diSA, not previously reported was also obtained (4.8mg of over 94% purity) through a similar separation of a different batch of QY using hydrochloric acid as the retainer and 10% dodecylamine as the ligand in the stationary phase. Two components that co-eluted (0.55g) in the 5g separation were separated when trifluoroacetic acid was used as the retainer and tetrabutylammonium hydroxide (an ion-pair reagent) as the ligand. The separation resulted in 20.7mg of 6',4-diSA, not previously reported, and 111.8mg of 8',5-diSA, both of over 98% purity. The isolated compounds were characterized by high-resolution mass spectrometry and proton nuclear magnetic resonance with correlated spectroscopy assignments. PMID- 19281994 TI - A rapid and reliable direct method for quantifying meat acylglycerides with monomode microwave irradiation. AB - A rapid methodology for direct analysis of meat acylglycerides is proposed. A transesterification is carried out in a microwave reactor consisting of a monomode oven using chlorotrimethylsilane (CTMS) and methanol. High-temperature gas chromatography was used to check the absence of underivatized acylglycerides. Whereas transesterification is complete after 30s at 90 degrees C in the microwave method, the reference method needs 2h to complete this process. Moreover, the CTMS-microwave method shows higher recoveries of individual saturated, monounsaturated and polyunsaturated fatty acids. No influence of microwave irradiation on the composition of the fatty acids was observed. PMID- 19281995 TI - Rapid and high-throughput purification of salvianolic acid B from Salvia miltiorrhiza Bunge by high-performance counter-current chromatography. AB - A large-scale purification of salvianolic acid B from Salvia miltiorrhiza Bunge is presented. The method development began with selection of the solvent system, then optimization of the operating parameters and ended up with linear scale-up from an analytical to a preparative instrument. Three factors were used for method optimization and scale-up estimation: purity, process throughput and process efficiency. Preparation was achieved using a two-phase solvent system comprising hexane-ethyl acetate-methanol-acetic acid-water (1:5:1.5:0.00596:5, v/v). This preparation yielded 475 mg of salvianolic acid B with a purity of 96.1% from 1.5 g of crude extract. The process throughput of crude was 2.23 g/h while process efficiency per gram of target compound was 0.769 g/h. Two factors process environmental risk factor and process evaluation factor were used for evaluation of the separation process. PMID- 19281996 TI - Two-dimensional liquid chromatography-ion trap mass spectrometry for the simultaneous determination of ketorolac enantiomers and paracetamol in human plasma: application to a pharmacokinetic study. AB - A bioanalytical method was developed for the simultaneous determination of paracetamol and ketorolac enantiomers in human plasma using two-dimensional liquid chromatography-mass spectrometry. Separation was first achieved in a reversed-phase C18 column by using a gradient solvent system consisting of 0.1% aqueous formic acid and acetonitrile (ACN). The effluent between 8.9 and 9.9 min, corresponding to phenacetin and racemic ketorolac peaks, was transferred to a polysaccharide-based chiral column (ChiralPak AD-RH) by using a six-port switching valve. Ketorolac enantiomers were subsequently separated on the chiral column using an isocratic mobile phase composed of ACN/0.1% formic acid 50:50 (v/v). The total run-time was less than 18 min. This innovative strategy prolongs the lifetime of chiral columns by avoiding damages due to the sample matrix. The detection was carried out with an ion trap mass spectrometer equipped with an electrospray ionisation source. The tested ranges were 0.05-20 microg/ml for paracetamol and 0.005-2 microg/ml for each ketorolac enantiomer. This method was fully validated and showed good performances in terms of trueness (80-110%) and precision (6.7-13.2%). The mean extraction recoveries were 60%, 72% and 76% for paracetamol, R-ketorolac and S-ketorolac, respectively. Finally, this procedure was successfully applied to a pharmacokinetic study. PMID- 19281997 TI - Surface chemistry of K-montmorillonite: ionic strength, temperature dependence and dissolution kinetics. AB - The surface chemistry of K-montmorillonite was investigated by potentiometric titrations conducted at 25, 50 and 70 degrees C and at ionic strengths of 0.001, 0.01 and 0.1 M KNO(3). Proton adsorption decreases with electrolyte concentration at all pHs. The pH of zero net proton charge (PZNPC) decreases from 8.1 to 7.6 when the ionic strength increases from 0.001 to 0.1 M. Temperature has a very small effect on surface charge. A constant capacitance model that accounts for protonation/deprotonation of aluminol and silanol edge sites and basal plane H(+)/K(+) exchange is used to fit the experimental data. H(+) and OH(-) adsorption to specific surface sites appear to account for the pH-dependence of the K-montmorillonite dissolution. PMID- 19281998 TI - Working memory and individual differences in mathematics achievement: a longitudinal study from first grade to second grade. AB - This longitudinal study examined the relationship between working memory and individual differences in mathematics. Working memory measures, comprising the phonological loop, the visuospatial sketchpad, and the central executive, were administered at the start of first grade. Mathematics achievement was assessed 4 months later (at the middle of first grade) and 1 year later (at the start of second grade). Working memory was significantly related to mathematics achievement in both grades, showing that working memory clearly predicts later mathematics achievement. The central executive was a unique predictor of both first- and second-grade mathematics achievement. There were age-related differences with regard to the contribution of the slave systems to mathematics performance; the visuospatial sketchpad was a unique predictor of first-grade, but not second-grade, mathematics achievement, whereas the phonological loop emerged as a unique predictor of second-grade, but not first-grade, mathematics achievement. PMID- 19281999 TI - Spatial variability associated with shifting land use: water quality and sediment metals in La Parguera, Southwest Puerto Rico. AB - Development in southwest Puerto Rico, as in many areas of the Caribbean, is outpacing the ability of upland flora, salt flats, and mangroves to capture sediments and intercept and transform nutrients. A comparative study to examine the effects of development on near-shore water quality in La Parguera, Puerto Rico, was initiated in 1998. Total suspended solids were significantly higher in the vicinity of developing areas compared to reference areas. Chlorophyll-a measurements near of the wastewater treatment plant averaged two times the level of other areas. The overall average concentrations of copper, chromium, nickel, and zinc in sediments collected from salt flats exceeded values reported to cause impairment of biological systems. Marine sediments near more developed locations had the highest metal concentrations, suggesting a greater transport in this area. Natural resources are the primary attraction in this area; therefore, protection of near-shore resources should receive greater attention in land use planning. PMID- 19282000 TI - [Cannabis. Risk/benefit]. PMID- 19282001 TI - [Hyperalgesia in patients treated with opioids with cancer pain]. PMID- 19282002 TI - [Risk factors of new fractures after vertebroplasty]. AB - PURPOSE: To evaluate the risk factors of new fractures after vertebroplasty (VP). PATIENTS AND METHOD: Prospective, non-randomized study including patients with acute osteoporotic fractures treated with VP. Baseline visit included clinical and densitometric data. At 30, 90 and 180 days, changes in clinical data and side effects (cement leakage and new fractures) were recorded. To establish the predictive factors of a new fracture, differences between the group of patients with new fractures (R1) and those without fractures (R0) were evaluated. RESULTS: Vertebroplasty was performed in 43 patients (82 vertebrae). Cement leakage into a disc appeared in 11 cases (11,5%) and 12 new fractures occurred in 9 patients. No statistical differences were detected between groups R1 and R0 in the following variables: sex, age, vitamin D levels, T-score, kyphosis angle, primary/secondary osteoporosis, preexisting fractures, number of treated vertebrae and amount of cement injected. A positive, statistical significant correlation, was established between cement leakage into a disk and incidence of adjacent new fractures (p<0.001). CONCLUSIONS: Cement leakage into a disc increases the risk of adjacent new fractures after vertebroplasty. PMID- 19282003 TI - [Functional anatomy of the vestibular nerve]. AB - The vestibular system detects head movements such as angular rotation, translation, and head position relative to gravity. It acts to stabilize the eyes and posture through subcortical reflexes. Its signals are also integrated at the cortical level to participate in the elaboration of a body scheme, used for different functions such as spatial orientation and motor control. The vestibular nerve shows a resting discharge rate that is modulated up or down according to head motion or position. Central functioning depends on the detection of an asymmetry between signals coming from a pair of peripheral sensors, one on either side. In pathological cases, unilateral peripheral dysfunction is interpreted by the central system as an asymmetry resulting from a change in head position leading to nystagmus, postural disturbances, and vertigo. The dysfunction can be either a deficit, such as observed in vestibular neuronitis, or hyperactivity such as observed in neurovascular compression syndrome of the VIIIth nerve. Anatomically, the VIIIth nerve has a long Root Entry Zone (REZ) that extends over 10mm before entering the brainstem. The VIIIth nerve is also physiologically close to numerous vessels at the pontocerebellar angle and internal auditory meatus. Therefore, vestibular syndrome resulting from neurovascular compression syndrome of the VIIIth nerve may exist, but it is very difficult to prove using radiological imagery. PMID- 19282004 TI - [Clinical and functional assessment of the vestibular nerve]. AB - Clinical and functional assessment of the vestibular nerve is fundamental in demonstrating vestibular signs and searching for associated otological and neurological signs. This may help orient topographic diagnosis toward central or peripheral syndrome and etiologic diagnosis. PMID- 19282005 TI - Methyl jasmonate induced accumulation of kalopanaxsaponin I in Nigella sativa. AB - Hydroponically cultivated Nigella sativa L. plants treated with methyl jasmonate (MeJA) showed a twelve-fold increase in levels of the monodesmosidic triterpene saponins alpha-hederin and kalopanaxsaponin I (KsI) in the leaves. We will demonstrate that these two saponins accounted for approximately 10% of the dry plant matter, of which 93% was KsI and 7% alpha-hederin. To address the molecular basis of saponin induction by MeJA, we cloned and characterized the beta-amyrin synthase gene (NsbetaAS1) encoding one of the key enzymes in triterpene saponin biosynthesis. As expected, NsbetaAS1 transcription was induced by MeJA and led to the production of beta-amyrin when over-expressed in yeast. PMID- 19282006 TI - [Diagnostic imaging departments' responses to terrorist attempts with multiple victims.]. AB - OBJECTIVES: To analyze whether the radiological management of seriously injured victims from the March 11 terrorist attempt was affected by the large number of victims treated at two hospitals in Madrid. To evaluate the organization for providing imaging services, detect failings, and propose a protocol for diagnostic imaging departments. MATERIAL AND METHODS: Two hundred and fifty one patients arrived at hospital A and 36 at hospital B. Both centers have emergency imaging areas and protocols for the treatment of patients with multiple trauma. We compared organizational aspects (classification, identification), material resources, human resources, healthcare resources (number and type of examinations), as well as the initial radiological management with the usual protocol and with the recommendations for incidents with multiple victims. RESULTS: In hospital A, patients' injuries were classified as severe (175) or minor (76); in hospital B, injuries were classified as extremely severe (13), severe (4), or minor (19). Additional staff were assigned to the emergency imaging areas in both hospitals. In hospital A, 62 portable plain-film radiographs, 39 ultrasonographic examinations, 25 cranial CT examinations, 6 cervical CT examinations, 2 chest CT examinations, and 2 abdominopelvic CT examinations were performed. In hospital B, 19 portable plain-film radiographs (74 in total), 9 ultrasonographic examinations, 17 cranial-chest-abdominopelvic CT examinations, 2 cervical CT examinations, 2 orbital CT examinations, and 2 CT examinations of the sinuses were performed. CONCLUSION: In both hospitals, each victim was managed as if he or she were the only patient. The discrepancies between the two hospitals were due to differences in the usual protocol for multiple trauma patients. In light of the organizational errors discovered, we propose a plan of action based on the identification and progressive activation of material and human resources until sufficient levels are achieved. PMID- 19282007 TI - [MR angiography follow-up of embolized cerebral aneurysms: interobserver agreement]. AB - OBJECTIVE: To determine the interobserver agreement in the interpretation of MR angiography (MRA) studies for surveillance of embolized intracranial aneurysms. To determine whether contrast administration improves interobserver agreement. MATERIAL AND METHODS: Two experienced neuroradiologists independently reviewed all follow-up MRA studies performed between July 2004 and December 2006 of cerebral aneurysms embolized with coils. All MRA studies included both unenhanced 3D time-of-flight (3D TOF) and contrast-enhanced MRA (CE-MRA) images. Studies were classified as: a) not assessable; b) complete occlusion; c) residual aneurysm. Interobserver agreement for unenhanced and enhanced MRA studies was determined using the kappa statistic. Kappa values were considered insignificant when<0.2, low when between 0.21 and 0.4, and moderate when between 0.41-0.6; values >0.6 were considered good agreement and >0.8 excellent agreement. Significance was set at p<0.005. RESULTS: We reviewed a total of 200 MRA studies (100 3D TOF studies and 100 CE-MRA studies) performed in 48 patients (25 women, 23 men) at 6, 12, and/or 24 months after embolization. Interobserver agreement was good in both 3D TOF and CE-MRA studies, although it was better in CE-MRA studies (kappa=0.660, p<0.001 and kappa=0.779, p<0.001, respectively). CONCLUSIONS: Interobserver agreement is good for follow-up MRA studies of embolized intracranial aneurysms. Gadolinium administration improves interobserver agreement. PMID- 19282008 TI - [Calcified cardiac thrombus in a patient with antiphospholipid syndrome]. AB - Antiphospholipid syndrome is a rare clinical entity that manifests through repeated thrombotic events in young patients without cardiovascular risk factors. The most frequent clinical features are repeated fetal losses and acute cerebral ischemic events. Cardiac involvement is less frequent. We present the case of a calcified thrombus in the right ventricle with extension to the pulmonary artery in a 30-year-old woman with a history of recurrent miscarriage who was diagnosed with antiphospholipid syndrome. PMID- 19282009 TI - [Case 6. Pseudoangiomatous stromal hyperplasia of the breast.]. PMID- 19282010 TI - [Comparative study of sentinel node biopsy in patients with multifocal breast carcinoma versus in those with unifocal breast carcinoma.]. AB - OBJECTIVE: To evaluate the results for sentinel node biopsy (SNB) in patients with multifocal breast cancer (MBC) in comparison to in those with unifocal breast cancer (UBC). PATIENTS AND METHODS: A total of 1535 prospective SNB (174 on patients with MBC) were performed at 9 hospitals. In most patients, Tc-99m albumin colloids were injected intraparenchymally into each tumoral focus for SNB. RESULTS: The overall identification rate was 93.8%; no differences between groups were observed (94.8% in MBC vs 93.4% in UBC). The mean number of sentinel nodes detected was 1.46, being higher in the MBC group than in the UBC group (1.58 vs 1.45; p=0.036). Extra-axillary sentinel nodes were found in 19.6%; extra axillary sentinel nodes were more common in the MBC group (23.4% vs 18.9%, ns) and in the internal mammary chain and in level III axillary lymph nodes. The incidence of sentinel node metastasis was 27.3% (29.1% MBC vs 26.7% UBC, ns), and the mean number of positive sentinel nodes was 0.42 in the MBC group vs 0.32 in the UBC group (p=ns). Axillary dissection identified the same rate of positive additional nodes (29.7%) in both groups. CONCLUSIONS: The diagnostic yield of SNB seems similar in MBC and UBC. In MBC, there appears to be a specific pattern of lymphatic drainage, with a higher number of sentinel nodes detected and probably a higher number of extra-axillary sentinel nodes. PMID- 19282011 TI - [Vascular-interventional radiology for hemodialysis: much more than a series of techniques in our catalogue (or from subspecialty to hyperspecialty)]. PMID- 19282012 TI - [Dual-source CT coronary angiography: image quality and optimal reconstruction interval]. AB - OBJECTIVE: To evaluate the effect of average heart rate on the quality of images of the coronary arteries and to determine the optimal reconstruction interval in nonselected patients undergoing DSCT coronary imaging. MATERIALS AND METHODS: We studied 28 consecutive subjects (26 men, 2 women; mean age 60+/-12 years) undergoing contrast-enhanced DSCT (Somatom Definition, Siemens) coronary angiography. Data sets were reconstructed in 5% steps from 30% to 80% of the R-R interval. Two blinded independent readers assessed the image quality of the whole coronary tree and of each coronary artery using a four-point scale (1: impossible to evaluate, 4: excellent quality). RESULTS: Mean heart rate during scanning was 73.1+/-14.8 bpm (range, 49-107 bpm). In the best reconstruction interval, excellent diagnostic image quality (score 4) was achieved for 98.8% (83/84) of coronary arteries. Very good interobserver agreement was observed for image quality rating (kappa=0.77) and for selection of the optimal reconstruction interval (kappa=0.86). Mean image quality score for the whole coronary tree was 2.84+/-0.31. No significant correlation was found between the average heart rate and the mean quality scores (r=0.108). No significant differences were found in the quality of images of the coronary arteries in systolic and diastolic reconstructions in each patient. CONCLUSIONS: DSCT makes it possible to obtain excellent coronary angiograms independent of the heart rate. Reconstructions can be obtained in either the diastolic or systolic phase without significant differences in image quality, even at high heart rates. PMID- 19282013 TI - [Case 8. Cardiac amyloidosis in magnetic resonance imaging.]. PMID- 19282014 TI - [Stroke in neonates and children]. AB - The clinical presentation, risk factors, causes, vital or functional prognosis, and acute management options for stroke occurring in neonates and children are specific, differing from those observed in young adults. Compared with the adult population, less is known about the epidemiology of stroke in the under-18 population where the disease could become more frequent because of advances in both neonatal resuscitation techniques for cerebral disorders and neuroimaging techniques enabling the diagnosis of small lesions. Clinical features are often delayed, especially in neonates, and unlike epilepsy or dystonia of the affected limb, which are frequent complications, aphasia is rather rare. The most frequent causes of stroke at the beginning of life are cardiac embolism, for ischemic stroke, and arteriovenous malformations, for intracerebral hemorrhage. Acute management at this age is specific. This article reviews the literature on the epidemiological and clinical features, the main causes, and the acute management guidelines of stroke occurring in newborn infants and children and highlights the need for neurologists to have comprehensive knowledge of this disease. PMID- 19282015 TI - The 5'-UTR of Turnip yellow mosaic virus does not include a critical encapsidation signal. AB - Turnip yellow mosaic virus (TYMV) RNA has two hairpins in the 5' untranslated region (UTR) with internal CC and CA mismatches that become protonated and are able to base pair at a pH near 5. The protonatable hairpins have previously been implicated as playing an important role in RNA encapsidation. We have examined the role of the 5'-UTR in the amplification and packaging of TYMV RNA using agroinfiltration of Chinese cabbage leaves to express various TYMV constructs with mutations affecting the 5'-UTR and the two hairpins. Mutations affecting the protonatable centers of the two hairpins, as well as deletion of one or both hairpins and deletion or mutation of the 17-nucleotide region upstream of the hairpins decreased viral amplification to varying extents (c. 10- to 1000-fold). However, in all these cases, the viral RNAs present in non-denaturing leaf extracts were predominantly ribonuclease resistant, indicative of encapsidation. These results show that, while the 5' hairpins are necessary for efficient amplification of TYMV, there appears to be no essential role for the 5'-UTR or its protonatable hairpins in the packaging of TYMV RNA. In a second set of experiments, it was demonstrated that TYMV can efficiently amplify in plants held in the dark, and that the progeny RNAs are efficiently encapsidated. Together, these observations argue for a revision of the model for TYMV encapsidation in which packaging occurs in low pH conditions that are generated by proton gradients produced by photosynthetic activity in the light and RNA packaging is dependent on the protonatable 5' hairpins. PMID- 19282016 TI - Synergism and negative interference during co-infection of tomato and Nicotiana benthamiana with two bipartite begomoviruses. AB - In Brazil, at least eight begomoviruses including Tomato rugose mosaic virus (ToRMV) and Tomato yellow spot virus (ToYSV) infect tomatoes. ToYSV symptoms in tomato and Nicotiana benthamiana appear earlier and are more severe compared to those of ToRMV. We investigated the role of several factors in this differential adaptation. To analyze infection kinetics, a single leaf was inoculated and subsequently detached after different periods of time. Viral DNA accumulation was quantified in plants, viral replication was analyzed in protoplasts, and tissue tropism was determined by in situ hybridization. Results indicate that ToYSV establishes a systemic infection and reaches a higher concentration earlier than ToRMV in both hosts. ToRMV negatively interferes with ToYSV during the initial stages of infection, but once systemic infection is established this interference ceases. In N. benthamiana, ToYSV invades the mesophyll, while ToRMV is phloem restricted. During dual infection in this host, ToYSV releases ToRMV from the phloem. PMID- 19282017 TI - Oxidation of enrofloxacin with conductive-diamond electrochemical oxidation, ozonation and Fenton oxidation: a comparison. AB - The treatment of enrofloxacin synthetic wastewaters using conductive-diamond electrochemical oxidation (CDEO), ozonation and Fenton oxidation has been studied. Results show that the three technologies can reduce the organic content of enrofloxacin synthetic wastewaters but with different performances. CDEO was the most efficient technology in terms of mineralization but not on COD removal, which was more efficiently achieved by ozonation. This indicates that ozonation is efficient in the breakage of the complex molecules but not on the removal of final carboxylic acids. The high initial efficiency in terms of oxidant-use obtained by Fenton oxidation evidences that it is very efficient in the removal of the enrofloxacin, although it rapidly leads to the formation of refractory compounds to the treatment. This indicates the significance of other oxidation mechanisms (e.g. coagulation) that enhance the results obtained by the expected hydroxyl-mediated oxidation. Ammonium ions were the primary product species in CDEO and nitrate ions in ozonation, whereas Fenton effluents contained similar amounts of both nitrogen ionic species. PMID- 19282018 TI - Progress and perspectives of sludge ozonation as a powerful pretreatment method for minimization of excess sludge production. AB - The treatment and disposal of excess sludge represents a bottleneck in wastewater treatment plants (WWTP) worldwide, due to environmental, economic, social and legal factors. The ideal solution to the problem of sludge disposal is to combine sludge reduction with the removal of pollution at the source. This paper presents an overview of the potential of ozonation in sludge reduction. The full-scale application of ozonation in excess sludge reduction is presented. Improvements in the biodegradability of the ozonated sludge were confirmed. The introduction of ozonation into activated sludge did not significantly influence effluent quality but improved the settling properties of the sludge. An operation with a suitable sludge wasting ratio seems to be necessary to prevent accumulation of inorganic and inert particles for long-term operation. Sludge ozonation to reduce excess sludge production may be economical in WWTP which have high sludge disposal costs and operational problems such as sludge foaming and bulking. The recommended ozone dose ranges from 0.03 to 0.05 g O(3)/g TSS, which is appropriate to achieve a balance between sludge reduction efficiency and cost. An effort to design and optimize an economic sludge reduction process is necessary. PMID- 19282019 TI - The aqueous photodegradation of fenitrothion under various agricultural plastics: implications for pesticide longevity in agricultural 'micro-environments'. AB - Plastic cladding is increasingly used in agriculture to create micro-environments to improve crop yield/growth. Many of these plastics can alter the solar light spectrum by inhibiting or reducing the transmittance of certain parts of the solar spectrum. In this study, we investigated the aqueous photolysis of fenitrothion, under a selection of different plastic films commonly used in agriculture. Three different grades of polyethylene film were used: 'standard', 'transparent' and 'opaque'. The transmittance of light wavelengths in the UV region (290-400 nm), measured with a spectroradiometer, was found to decrease in the order of transparent>standard>opaque. Fenitrothion, an organophosphorothioate insecticide possesses molar absorptivity in the solar wavelength range of 290-400 nm. Aqueous first order degradation rate constants for fenitrothion determined over a 12 h period were found to be considerably less for those experiments conducted under the standard and opaque plastic films, compared to the transparent film and no-film control. The experiments were conducted in an Atlas Suntest solar simulator using a UV-filtered Xenon arc lamp to simulate sunlight. The first order half-life for fenitrothion was 100 and 250 h under the standard and opaque films, respectively, compared to approximately 10 h for the transparent film and no-film experiments. Our results suggest that pesticide longevity could be greatly extended within these plastic micro-environments, especially for those chemicals which may degrade/transform via photolytic or photochemical pathways. PMID- 19282020 TI - Sorption of naphthalene and 1-naphthol by biochars of orange peels with different pyrolytic temperatures. AB - Biochars, derived from biomass, are increasingly recognized as an environmental friendly sorbent to abate organic pollutants. Sorption variations of biochars with their pyrolytic temperatures are evaluated. Nine biochars of orange peels with different pyrolytic temperatures (150-700 degrees C, referred as OP150 OP700) were characterized via elemental analysis, BET-N(2) surface area, and Fourier transform infrared spectroscopy. Sorption of naphthalene and 1-naphthol by the biochars in water are compared. Sorption isotherms varied from linear to Freundlich with increasing pyrolytic temperature. The respective contributions of adsorption and partition to total sorption were correlated with biochars' structural parameters. For OP150-OP600, sorption of 1-naphthol was significantly larger than naphthalene due to the former owning additional specific interactions. For 1-naphthol with high concentrations, the OP200 exhibited the maximal sorption capacity due to its largest partition and high adsorption among nine biochars. For 1-naphthol with low concentrations and naphthalene, the OP700 displayed the maximal sorption capacity. These observations provide a reference to the use of biochars as engineered sorbents for environmental applications. PMID- 19282021 TI - Hydrolysis and H2O2-assisted UV photolysis of 3-chloro-1,2-propanediol. AB - 3-Chloro-1,2-propanediol (3-MCPD) is a chlorinated alcohol that is often formed as a by-product in the manufacturing of food products. In addition, 3-MCPD may be a disinfection by-product from wastewater treatment by chlorine and may be present in drinking waters from purification plants using epichlorohydrin-linked cationic polymer resins as flocculants. Due to concerns about the toxicity of 3 MCPD and its potential presence in water samples, the removal of 3-MCPD from water should be addressed and examined. For the first time a systematic examination of the removal of 3-MCPD via hydrolysis and photolysis processes is presented. 3-MCPD is shown to undergo hydrolysis at near neutral pH values, but at much slower rates than can be obtained by UV/H2O2 processes. 3-MCPD does not undergo rapid direct photolysis. Re-evaluation of temperature and pH dependent hydrolysis rate data indicates that hydrolysis is first order with respect to [OH(-)]. PMID- 19282022 TI - Dissection of soft-preserved cadavers in the training of gynaecological oncologists: report of the first UK workshop. AB - OBJECTIVE: To assess the surgical anatomy knowledge of gynaecological oncology (GO) trainees and to evaluate the impact of a cadaveric dissection course on postgraduate surgical training. METHODS: An intensive 3-day cadaveric dissection course with illustrated lectures and supervised dissection, with a multiple choice questionnaire (MCQ) on surgical anatomy at the beginning and end of the course was organised in the Anatomy Facility of a London Medical School. Each cadaver was embalmed with a mixture of alcohol, phenol and glycerol ("soft preserved") rather than fixed in formalin, to more closely preserve in vivo conditions of the body. There were ten dissecting delegates, two per cadaver. The delegates dissected the abdomen and pelvis with the emphasis on surgical approaches rather than the classical descriptive anatomy approaches. Delegates also completed a course evaluation. RESULTS: Without negative marking, the mean initial MCQ score was 57%, and final mean score 64%. With negative marking, the mean initial score was 43%, and mean final score 53%. Delegates rated the course highly, would recommend it to other trainees and considered that such a course should be incorporated into subspecialty training. CONCLUSION: The surgical anatomy knowledge of subspecialty trainees was weak but improved as a result of the dissection course. The most positive finding was the course evaluation. Postgraduate surgical training in GO would likely be enhanced by, and arguably requires, cadaveric dissection. "Soft-preserved" rather than formalin-fixed cadavers should be used. PMID- 19282023 TI - Postpartum anestrus in dairy cattle. AB - Fertility of the postpartum period is negatively influenced by the incidence of anestrus. The latter condition is characterized by the absence of estrous behavior, which may be an indication of suboptimal conditions (e.g., inadequate peripartum nutrition) or pathologic conditions (e.g., chronic debilitating diseases or uterine and ovarian diseases). Although initiation of ovarian follicular growth in the postpartum period is generally not affected, subsequent development (deviation) and the fate of the dominant follicle are the primary factors that affect reestablishment of ovarian cyclicity. Anestrus can be classified based on the three functional states of follicular development; that is, follicle emergence, deviation, and ovulation. Prevention of anestrus is preferable to treatment and can be achieved in part by maintaining a healthy periparturient period. To better understand the etiology of anestrus and its prevention, research is urgently needed in the following three areas: the role of peripartum disease conditions that influence reproduction, genes involved in ovulation, and the influence of proteins (e.g., leptin) that appear to be important links between metabolic signals and the neuroendocrine axis. PMID- 19282024 TI - Effect of glutamine on post-thaw motility of bull spermatozoa after association with LDL (low density lipoproteins) extender: preliminary results. AB - Glutamine has been used in the composition of semen extenders in several species, but never in the bull. The aim of our study is to demonstrate the cryoprotective role of glutamine for freezing bovine semen and to determine concentration of the latter to improve the motility and trajectory characteristics of spermatozoa. Three experiments were undertaken with 21 ejaculates from three different bulls. In the first experiment, glutamine was added to 40, 80, and 120 mM of basic medium (BM) which consisted of Tris+glycerol 6.4% (v/v). In the second experiment glutamine was added to the 8% low density lipoprotein (LDL) diluent at 40, 80, and 120 mM. In the third experiment, the best concentration of glutamine was determined; this was then added to the LDL extender at 10, 20, 30, and 40 mM. The semen was diluted then frozen in the different media. Motility parameters were assessed using an image analyser following thawing. Experiment 1 demonstrated that glutamine had a cryoprotective effect; at 40 mM it gave superior motility parameters to those obtained with the basic medium (p<0.05). Experiment 2 demonstrated that the combination of LDL-glutamine 40 mM and 80 mM did not improve motility and even deteriorated it in comparison with the glutamine-free LDL extender. Experiment 3 demonstrated that the addition of 10mM of glutamine to the LDL medium lead to a significant improvement (p<0.05) in the motility of bull spermatozoa and could be used to improve bovine semen extenders. PMID- 19282025 TI - The war on drugs: a devastating public-policy disaster. PMID- 19282027 TI - World Kidney Day: hypertension and chronic kidney disease. PMID- 19282026 TI - Graft-versus-host disease. AB - Haemopoietic-cell transplantation (HCT) is an intensive therapy used to treat high-risk haematological malignant disorders and other life-threatening haematological and genetic diseases. The main complication of HCT is graft-versus host disease (GVHD), an immunological disorder that affects many organ systems, including the gastrointestinal tract, liver, skin, and lungs. The number of patients with this complication continues to grow, and many return home from transplant centres after HCT requiring continued treatment with immunosuppressive drugs that increases their risks for serious infections and other complications. In this Seminar, we review our understanding of the risk factors and causes of GHVD, the cellular and cytokine networks implicated in its pathophysiology, and current strategies to prevent and treat the disease. We also summarise supportive care measures that are essential for management of this medically fragile population. PMID- 19282028 TI - The FPR2-specific ligand MMK-1 activates the neutrophil NADPH-oxidase, but triggers no unique pathway for opening of plasma membrane calcium channels. AB - Human neutrophils express formyl peptide receptor 1 and 2 (FPR1 and FPR2), two highly homologous G-protein-coupled cell surface receptors important for the cellular recognition of chemotactic peptides. They share many functional as well as signal transduction features, but some fundamental differences have been described. One such difference was recently presented when the FPR2-specific ligand MMK-1 was shown to trigger a unique signal in neutrophils [S. Partida Sanchez, P. Iribarren, M.E. Moreno-Garcia, et al., Chemotaxis and calcium responses of phagocytes to formyl peptide receptor ligands is differentially regulated by cyclic ADP ribose, J. Immunol. 172 (2004) 1896-1906]. This signal bypassed the emptying of the intracellular calcium stores, a route normally used to open the store-operated calcium channels present in the plasma membrane of neutrophils. Instead, the binding of MMK-1 to FPR2 was shown to trigger a direct opening of the plasma membrane channels. In this report, we add MMK-1 to a large number of FPR2 ligands that activate the neutrophil superoxide-generating NADPH oxidase. In contrast to earlier findings we show that the transient rise in intracellular free calcium induced by MMK-1 involves both a release of calcium from intracellular stores and an opening of channels in the plasma membrane. The same pattern was obtained with another characterized FPR2 ligand, WKYMVM, and it is also obvious that the two formyl peptide receptor family members trigger the same type of calcium response in human neutrophils. PMID- 19282029 TI - Estimating the prevalence of myelodysplastic syndromes in patients with unexplained cytopenias: a retrospective study of 322 bone marrows. AB - Although unexplained anemia is common in the elderly, the prevalence of MDS is poorly defined. We reviewed 2267 bone marrows reviewed at our center between the years 2002 and 2005. Of these, 322 met our criteria for inclusion (14%). Seventy three patients (22.6%) had a confirmed diagnosis of MDS and 32 (9.9%) had suspected MDS. Confirmed or suspected MDS was more likely in patients aged >65 (31.5% and 11%, respectively). Age, MCV, LDH and RDW were independently predictive of MDS. Extrapolation of our findings to the elderly anemic individuals in the community suggests MDS prevalence may be higher than previously postulated. PMID- 19282030 TI - ABL deletion without associated BCR-ABL in precursor B-cell acute lymphoblastic leukemia. PMID- 19282031 TI - Authors' response to Vieth (2008): legal and psychological support for the NICHD Interviewing Protocol. PMID- 19282032 TI - Phylogenetic analysis of human bocavirus (HBoV) detected from children with acute respiratory infection in Japan. PMID- 19282033 TI - How suicidal bipolar patients are depends on how suicidal ideation is defined. AB - BACKGROUND: Suicidal ideation indicates risk for suicidal acts. How different definitions and measures for suicidal ideation influence its prevalence, correlates and predictive validity among bipolar disorder (BD) patients is unknown. METHODS: Among the 191 BD patients in the Jorvi Bipolar Study (JoBS), suicidal ideation at baseline was measured using the Scale for Suicidal Ideation (SSI), Hamilton Depression Scale (HAM-D) item 3 and Beck Depression Inventory (BDI) item 9 and by asking whether patients had seriously considered suicide. The predictive value of different definitions of ideation on suicide attempts during a six-month follow-up was investigated. RESULTS: Altogether 74% of patients had suicidal ideation as defined in at least one of the above-mentioned ways, but only 29% met the criteria for all ways; agreement between definitions ranged from low to moderate (kappa coefficient 0.15 to 0.70). The correlates of suicidal ideation overlapped, but were not identical. Of the measures investigated, a baseline SSI score >or=8 had the best combination of sensitivity (0.81) and specificity (0.69) and a positive predictive value (PPV) of 32% for an attempted suicide during follow-up. LIMITATIONS: All plausible measures for suicidal ideation could not be investigated. CONCLUSIONS: Who is classified as having suicidal ideation depends strongly on the definition and means of measurement among BD patients. Different measures for ideation have the potential to cause inconsistency when correlates of suicidal ideation are investigated. For clinically predicting suicide attempts during the next few months, an SSI score >or=8 may best combine sensitivity and specificity. PMID- 19282034 TI - Suspiciousness and low self-esteem as predictors of misattributions of anger in schizophrenia spectrum disorders. AB - While it is widely recognized that many with schizophrenia have significant difficulties in correctly identifying the emotions of others, less is known about the causes and correlates of particular forms of misattribution, including mistakenly seeing anger in others. One possibility is that persons with high levels of suspiciousness and low levels of self-esteem are at risk to attribute their poor feelings about themselves to the malice of others. To explore this possibility, we identified 52 persons with a schizophrenia spectrum disorder who made significant numbers of errors on the Bell-Lysaker Emotional Recognition Test. We then performed a cluster analysis based on measures of suspiciousness from the Positive and Negative Syndrome Scale and self-esteem from the Rosenberg Self-Esteem Schedule, and found the following four groups: a) High Suspiciousness/High Self-Esteem; b) Mild Suspiciousness/High Self-Esteem; c) High Suspiciousness/Low Self-Esteem; and d) Minimal Suspiciousness/Low Self-Esteem. Comparisons between groups revealed that as predicted the High Suspiciousness/Low Self-Esteem group made significantly more misattributions of anger than other groups, even when levels of depression were controlled for statistically. Implications for addressing the misattributions of anger in schizophrenia are discussed. PMID- 19282035 TI - Evaluating the reliability of multiple assessments of PTSD symptomatology: multiple examiners, one patient. AB - The objective of this investigation was to assess the inter-examiner reliability of PTSD symptomatology by 12 clinical examiners who evaluated independently a single-case Vietnam-Era veteran, using videotaped clinician interviews with The Clinician Administered PTSD Scale-1 (CAPS-1). A second patient was utilized for cross-validation purposes. Data were analyzed using a specially designed Kappa statistic. In previous reliability assessments of the CAPS-1, a pair of examiners assessed multiple patients, and demonstrated evidence of high reliability and validity. As in previous reliability assessments, reliability was assessed both for frequency and intensity of PTSD symptomatology in both patients. Results indicated that the reliability levels of the CAPS-1 were consistently and almost exclusively in the excellent to perfect levels of inter-examiner agreement, as based upon both global assessments and on a symptom-by-symptom basis. The results of this investigation are interpreted in the broader framework of their applicability to assessing inter-examiner agreement in clinical trials or other large multi-site studies. PMID- 19282036 TI - Prevalence of malocclusion among mouth breathing children: do expectations meet reality? AB - OBJECTIVE: The aim of this study was to report epidemiological data on the prevalence of malocclusion among a group of children, consecutively admitted at a referral mouth breathing otorhinolaryngological (ENT) center. We assessed the association between the severity of the obstruction by adenoids/tonsils hyperplasia or the presence of allergic rhinitis and the prevalence of class II malocclusion, anterior open bite and posterior crossbite. METHODS: Cross sectional, descriptive study, carried out at an Outpatient Clinic for Mouth Breathers. Dental inter-arch relationship and nasal obstructive variables were diagnosed and the appropriate cross-tabulations were done. RESULTS: Four hundred and one patients were included. Mean age was 6 years and 6 months (S.D.: 2 years and 7 months), ranging from 2 to 12 years. All subjects were evaluated by otorhinolaryngologists to confirm mouth breathing. Adenoid/tonsil obstruction was detected in 71.8% of this sample, regardless of the presence of rhinitis. Allergic rhinitis alone was found in 18.7% of the children. Non-obstructive mouth breathing was diagnosed in 9.5% of this sample. Posterior crossbite was detected in almost 30% of the children during primary and mixed dentitions and 48% in permanent dentition. During mixed and permanent dentitions, anterior open bite and class II malocclusion were highly prevalent. More than 50% of the mouth breathing children carried a normal inter-arch relationship in the sagital, transversal and vertical planes. Univariate analysis showed no significant association between the type of the obstruction (adenoids/tonsils obstructive hyperplasia or the presence of allergic rhinitis) and malocclusions (class II, anterior open bite and posterior crossbite). CONCLUSIONS: The prevalence of posterior crossbite is higher in mouth breathing children than in the general population. During mixed and permanent dentitions, anterior open bite and class II malocclusion were more likely to be present in mouth breathers. Although more children showed these malocclusions, most mouth breathing children evaluated in this study did not match the expected "mouth breathing dental stereotype". In this population of mouth breathing children, the obstructive size of adenoids or tonsils and the presence of rhinitis were not risk factors to the development of class II malocclusion, anterior open bite or posterior crossbite. PMID- 19282037 TI - Trophic functions of nucleotides in the central nervous system. AB - In addition to short-term effects, one of the fundamental roles of extracellular nucleotides in the central nervous system involves long-term trophic effects. Physiological outcomes include neurogenesis, neuronal differentiation, glial proliferation, migration, growth arrest and apoptosis. Nucleotides exert these functions via P2-receptor-mediated mechanisms that can also interact with polypeptide-growth-factor-mediated or integrin-mediated signaling pathways. In addition, pathogenic roles for extracellular nucleotides in response to central nervous system injury including trauma and ischemia have been observed after the release of nucleotides by damaged and dying cells and in the development of neuropathic and inflammatory pain. Here, we illuminate the contribution of extracellular nucleotides to the development, growth, cellular plasticity and death of neural cells and the mechanisms regulating these trophic effects. PMID- 19282038 TI - Histological biomarkers in liver and gills of juvenile Solea senegalensis exposed to contaminated estuarine sediments: a weighted indices approach. AB - Young juvenile Solea senegalensis were exposed to three sediments with distinct contamination profiles collected from a Portuguese estuary subjected to anthropogenic sources of contamination (the Sado estuary, western Portugal). Sediments were surveyed for metals (cadmium, chromium, copper, nickel, lead and zinc), a metalloid (arsenic) and organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls and a pesticide, dichloro-diphenyl trichloroethane plus its metabolites), as well as total organic matter, redox potential and particle fine fraction. The fish were exposed to freshly collected sediments in a 28-day laboratorial assay and collected for histological analyses at days 0 (T(0)), 14 (T(14)) and 28 (T(28)). Individual weighted histopathological indices were obtained, based on presence/absence data of eight and nine liver and gill pathologies, respectively, and on their biological significance. Although livers sustained more severe lesions, the sediments essentially contaminated by organic substances caused more damage to both organs than the sediments contaminated by both metallic and organic contaminants, suggesting a possible synergistic effect. Correlation analyses showed that some alterations are linked, forming distinctive histopathological patterns that are in accordance with the severity of lesions and sediment characteristics. The presence of large eosinophilic bodies in liver and degeneration of mucous cells in gills (a first-time described alteration) were some of the most noticeable alterations observed and were related to sediment organic contaminants. Body size has been found to be negatively correlated with histopathological damage in livers following longer term exposures. It is concluded that histopathological indices provide reliable and discriminatory data even when biomonitoring as complex media as natural sediments. It is also concluded that the effects of contamination may result not only from toxicant concentrations but also from their interactions, relative potency and sediment characteristics that ultimately determine bioavailability. PMID- 19282039 TI - A brief clinical tool to assess physical function: the mini-physical performance test. AB - The aim was to develop a brief physical performance assessment tool that can be reliably used to detect physical impairment in older adults with and without mild dementia. Scores on the 9-item physical performance test (PPT) from non-demented participants were used to develop and validate the 4-item mini-PPT. The validated mini-PPT was then used to predict total PPT score and functional physical status in participants with mild dementia. Receiver operating curve (ROC) analyses were used to generate a cutoff score that classifies participants as functional vs. not functional. The setting was in the Alzheimer's Disease Research Center (Washington University). A total of 1199 participants met inclusion criteria: 574 non-demented participants, 436 with very mild dementia, measured by the clinical dementia rating (CDR)=0.5 and 189 with mild dementia (CDR=1). The mean age of the sample was 76.4 years, mean educational attainment was 14 years, 58% were women, and 11% were African American. A 4-item scale, the mini-PPT, was developed (based on the results of multiple regression analyses and clinical meaningfulness) that highly correlated with total PPT score (r=0.917, p<0.0001) in the non-demented sample. The correlation of the mini-PPT with total PPT was 0.90 among those with very mild, and 0.91 among those with mild dementia. Using the ROCs, a cutoff score of 12 correctly classified at least 85% of non-demented and demented persons. The 4-item mini-PPT is highly correlated with the 9-item PPT in non demented and mildly demented persons. This brief tool may be useful in detecting early physical impairment in the clinical setting. PMID- 19282040 TI - Hydrotherapy after total hip arthroplasty: a follow-up study. AB - The aim of the study was to evaluate the subjective functional outcome of total hip arthroplasty (THA) in patients who underwent hydrotherapy (HT) 6 months after discharge. A prospective randomized study was performed on 70 elderly inpatients with recent THA, who completed a rehabilitation program. After randomization, 33 of them were treated in conventional gyms (no-hydrotherapy group=NHTG) and 31 received HT (hydrotherapy group=HTG). Interviews with the Western-Ontario MacMasters Universities Osteoarthritis Index (WOMAC) were performed at admission, at discharge and 6 months later. Kruskal-Wallis, Mann-Whitney and Wilcoxon tests were applied for statistical analysis. Both groups improved. Pain, stiffness and function were all positively affected. Statistical analysis indicated that WOMAC sub-scales were significantly lower for all patients treated with HT. The benefits at discharge still remained after 6 months. We conclude that HT is recommended after THA in a geriatric population. PMID- 19282041 TI - Right atrial myxoma presenting as syncope due to pulmonary embolism in a previously asymptomatic patient. AB - Tumours of the heart are rare, and in the vast majority of cases are myxomas that have a benign profile. According to their location, their clinical manifestations differ but there are cases that are completely asymptomatic [M. Yuce, S. Dagdelen, M. Ergelen, N. Eren, N. Caglar, A huge obstructive myxoma located in the right heart without causing any symptom. Int J Cardiol 2007;114:405-406.], found only post-mortem. Herein we describe the case of a 45-year-old female patient previously asymptomatic who presented with syncope due to a large right atrial myxoma with extracardiac localization as embolic phenomenon in the pulmonary artery tree. PMID- 19282042 TI - Evidence-based prescribing and adherence to antiplatelet therapy--how much difference do they make to patients with atherothrombosis? AB - Antiplatelet therapy is vital for the secondary prevention of cardiovascular events in patients with atherothrombosis (acute coronary syndromes, cerebrovascular disease and peripheral arterial disease). Numerous international guidelines provide evidence-based recommendations about the optimal use of antiplatelet agents in these disease states. This article reviews the published literature in order to firstly quantify the extent to which guideline recommendations for antiplatelet agents are taken up by prescribing physicians, and the degree to which patients adhere to prescribed treatments. It then goes on to examine the consequences of non-adherence and explores ways in which adherence may be improved from the perspective of both the physician and the patient. PMID- 19282043 TI - Association between the use of non-antiarrhythmic drugs and postoperative atrial fibrillation. AB - PURPOSE: To evaluate the association between statins, N-acetylcysteine (NAC) and angiotensin converting enzyme inhibitors (ACEIs) or angiotensin receptor blockers (ARBs) and postoperative atrial fibrillation (AF). METHODS: A total of 590 patients undergoing coronary artery bypass and/or valve surgery were studied. An AF episode lasting >5 min during hospitalization was accepted as endpoint. RESULTS: AF rate was 18% (n=106). Multivariable positive predictors of AF included age >= 65, left atrial diameter >= 45 mm, cross clamp time; negative predictors included use of left internal mammarian artery, NAC, and ACEIs or ARBs. CONCLUSIONS: ACEIs or ARBs and NAC is associated with low incidence of postoperative AF, however, this association was not found with statin or combined use these agents. PMID- 19282044 TI - Exopolysaccharide production by lactose-hydrolyzing bacteria isolated from traditionally fermented milk. AB - With increasing consumer demands for safer, healthier and more natural products, bacterially produced exopolysaccharides (EPSs) are becoming a viable option as an additive in designer-type foods. Fresh milk samples from cattle and sheep were collected from informal settlements in South Africa. After a three day incubation period at 25 degrees C, 550 bacterial strains were isolated and evaluated for EPS production from lactose as sole carbon source. Strains producing EPS on lactose were identified to species level with 16S rRNA gene sequencing and encompass 11 Gram-positive and 6 Gram-negative bacteria. EPS production was assigned for the first time to members of the species Staphylococcus hominis and Enterococcus lactis, and also to apparently novel species of the genera Sphingomonas and Acinetobacter. The polymers consisted mainly out of galactose and glucose, while a few isolates also incorporated rhamnose. Isolates produced diverse biopolymers as seen by significant differences in monomer ratios. PMID- 19282045 TI - Autonomic nerve fiber function and bone mineral density in individuals with type 1 diabetes: a cross-sectional study. AB - AIMS: Experimental models support a role for the sympathetic nervous system in bone metabolism. Beta-adrenoreceptors have been demonstrated on osteoblast-like cells. Beta-blocker use is associated with higher bone mineral density (BMD) in some epidemiologic studies. The aim of this study was to determine if measures of the autonomic nervous system (ANS) were associated with reduced BMD in type 1 diabetes. METHODS: We studied 66 individuals with type 1 diabetes. Dual-energy X ray absorptiometry was used to measure BMD and bone mineral content (BMC). Measures of heart rate variability (HRV) (e.g., power spectral analysis, mean circular resultant) and circulating norepinephrine levels were used to evaluate autonomic function. RESULTS: BMD/BMC were examined by gender for individuals with a normal versus elevated low-frequency (LF) band (expressed in normalized units). No significant differences in BMD/BMC were shown for individuals with an elevated LF band. Direct-entry linear regression models with BMD/BMC as the dependent variable were performed. Duration, gender, BMI, HbA(1c), insulin dose/kg, activity energy expenditure, and measures of HRV were entered as potential independent variables. No measures of HRV, or norepinephrine, were independently associated with BMD/BMC. CONCLUSIONS: The ANS, as assessed by measures of HRV, does not appear to impact BMD/BMC in type 1 diabetes. PMID- 19282046 TI - Increased serum levels of ghrelin at diagnosis mediate body weight loss in non small cell lung cancer (NSCLC) patients. AB - OBJECTIVE: Ghrelin is an orexigenic peptide implicated in body weight regulation, while cachexia is a multifactorial effect of non-small cell lung cancer (NSCLC) presented in patients with advanced disease. The aim of this study was to detect the role of ghrelin in cachexia and systemic inflammation of advanced NSCLC patients as well as its role as a diagnostic and prognostic tool. METHODS: Ghrelin serum levels were measured in 101 inoperable NSCLC patients before receiving any therapy (75 patients with weight loss and 26 without weight loss) and 60 healthy control volunteers. Epidemiological, anthropometrical and laboratory data were assessed for all participants (patients and healthy volunteers). RESULTS: NSCLC patients presented significantly higher ghrelin serum levels than healthy individuals, adjusted for sex, age and BMI (0.5+/-0.4 ng/ml vs. 0.4+/-0.3 ng/ml, P<0.001). NSCLC patients with weight loss presented significantly increased ghrelin serum levels (0.56+/-0.24 ng/ml vs. 0.52+/-0.44 ng/ml, P=0.017), compared to NSCLC patients without weight loss. CONCLUSIONS: Ghrelin serum levels are significantly increased in NSCLC patients, mainly in the subgroup of patients diagnosed with cachexia, indicating a possible implication in the pathogenesis of lung cancer. Further studies are needed to determine its potential role as predictive and prognostic marker. PMID- 19282047 TI - Signs of sex: what we know and how we know it. AB - The predominance of sexual reproduction indicates that it must confer profound benefits, considering its significant costs relative to asexuality. However, definitively determining whether a lineage engages in sex is often complicated by the potential for cryptic sex, especially among unfamiliar organisms. Here we consider the strengths and weaknesses of various molecular- and organismal-based approaches for recognizing signs of sex and describe their applications and relevance to evolutionary biology. We review recent studies that use these methods; some analyses even dispute several 'ancient' asexual taxa, and suggest they are recently derived or might be covertly sexual. More broadly, a better understanding of which organisms have sex and how they do it will deepen our understanding of the distribution, maintenance and evolution of sexual reproduction. PMID- 19282048 TI - Does patient-specific risk adjustment lead to different conclusions on the occurrence of surgical wound infections? PMID- 19282049 TI - Patients and the public: knowledge, sources of information and perceptions about healthcare-associated infection. AB - Statutory bodies provide information about healthcare-associated infection (HCAI) in the UK. Information is also available on National Health Service trust websites. Opinion polls demonstrate that fear of developing HCAI, especially methicillin-resistant Staphylococcus aureus, is the single greatest concern of people contemplating healthcare. We undertook a literature review to determine lay knowledge of HCAI, sources of information and perceptions of the risks. Twenty-two studies met the inclusion criteria. Of these, nine explored knowledge and perceptions as the primary research aim. The remainder consisted of a heterogeneous assortment of works comparing the knowledge and perceptions of different groups, their experiences of being infected or colonised and/or isolated. In all accounts, lay people expressed anxiety about the risks and consequences of HCAI. The most frequently reported source was the media, which has been blamed for sensationalist and inaccurate accounts. Lay people do not appear to access credible sources of information, or, if they do access them, are unable to understand their messages. Organisations that provide patient-focused information about HCAI are generic in scope, so that obtaining specific information may take time and effort to locate. Research is necessary to explore the acceptability, comprehensibility and accessibility of lay sources of information about HCAI and to find ways of readjusting risk perceptions to realistic levels in order to provide sensible levels of reassurance to those about to undergo healthcare. PMID- 19282050 TI - Assessing staff knowledge about Clostridium difficile diarrhoea. PMID- 19282051 TI - Epidemiological investigation after hospitalising a case with pandrug-resistant Acinetobacter baumannii infection. AB - Pandrug-resistant Acinetobacter baumannii (PDRAB) emerged in Taiwan in the early 2000s but was not identified in the Children's Hospital (Hospital A) until March 2005 when a patient was transferred from a respiratory care hospital (Hospital B). PDRAB was recovered from an eye swab taken on admission; once aware of the culture result, in addition to implementing infection control precautions, an epidemiological investigation was conducted in both hospitals. A total of 212 specimens were taken from 30 hospital inpatients (seven in Hospital A, 23 in Hospital B), clinical equipment and ward environment. Thirteen (15.5%) of 84 specimens obtained from Hospital A and 23 (18%) of 128 specimens obtained from Hospital B were positive for A. baumannii; of these, six isolates from two patients and clinical equipment in Hospital A and five from three patients in Hospital B were PDRAB. One patient stayed in both hospitals, and had one PDRAB isolate detected at each. Of the 36 A. baumannii isolates, there were nine IRS PCR (infrequent restriction site-polymerase chain reaction) patterns, 12 PFGE (pulsed-field gel electrophoresis) patterns and six antibiogram patterns identified. Twenty-five isolates belonged to a major IRS-PCR type (four PFGE patterns) and presented with either pandrug resistance (all 11 PDRAB isolates clustered in this type) or multidrug resistance (only susceptible to imipenem). A. baumannii is an ubiquitous organism that can be isolated from patients and their equipment. A clone of A. baumannii with multi- or pandrug resistance was circulating in both hospitals. PMID- 19282052 TI - Development of a method to measure bacterial transfer from hands. AB - A method was developed to investigate the transfer of bacteria from the hands of healthcare workers (HCWs). The method involved standardised hand contact between the HCW and a recipient wearing sterile gloves, followed by sampling of the bare hands of the HCW and the gloved hands of the recipient by the glove juice method. The duration of contact, degree of friction and dryness of the hands could be varied. We investigated the applicability of the method for measuring transfer from hands artificially contaminated with Escherichia coli as well as from naturally contaminated hands following a 30s contact time with moderate friction and dry hands. Only a small proportion of bacteria on donor hands was recovered from the recipient: 0.15% for E. coli and 0.07% for natural hand flora. A smaller proportion of E. coli was recovered from bare skin compared with gloves, suggesting reduced survival of bacteria as a result of contact with natural skin. We suggest that these data are clinically relevant, and may indicate low transfer of bacteria during short contact with dry hands. This method is suitable to investigate the effect of potential risk factors for ineffective hand hygiene and the effect of hand hygiene procedures on contact transmission in clinical studies with large numbers of HCWs. PMID- 19282053 TI - Can hospital computers be disinfected using a hand-held UV light source? PMID- 19282054 TI - Public perception of Clostridium difficile. PMID- 19282055 TI - Varicella zoster exposure on paediatric wards between 2000 and 2007: safe and effective post-exposure prophylaxis with oral aciclovir. AB - Varicella zoster virus is highly contagious and can cause serious complications in immunocompromised patients. To prevent people exposed to the virus from developing secondary varicella we have used oral aciclovir as post-exposure prophylaxis (PEP) since 2000. Between 2000 and 2007, there were 11 unexpected occurrences of varicella and 11 unexpected occurrences of zoster in our paediatric wards. There were 174 contacts, 131 exposed to varicella and 43 exposed to zoster. A total of 163 (94%) received PEP and 11 (6%) did not. The rates of secondary infection among contacts given prophylaxis with aciclovir only were 2.1% (3/141) for all contacts and 1.3% (1/76) for immunocompetent contacts. The rate of secondary infection among contacts not given PEP was significantly higher (18%, 2/11) (P<0.05). No adverse events due to PEP were reported. We conclude that oral aciclovir PEP following exposure to VZV on paediatric wards is both safe and effective. PMID- 19282056 TI - Role of contaminated aspiration tubes in nosocomial outbreak of Klebsiella pneumoniae producing SHV-2 and CTX-M-15 extended-spectrum beta-lactamases. AB - Klebsiella pneumoniae resistant to ceftazidime was isolated from ten neonates hospitalised between February and March 2006 in two Antananarivo hospitals, Madagascar. The main environmental source, for one hospital in particular, was the liquid used to rinse aspiration tubes in the paediatric wards. The risk of contamination from aspiration tubes is very high in the hospitals of Antananarivo since tap water used to rinse the tubes is not regularly changed. Phenotypical (biotyping and antibiotyping) and genotypical (pulsed-field gel electrophoresis) analysis of all the clinical isolates indicated that nine cases were due to a single clone. This clone carried the genes encoding SHV-2 and CTX-M-15 beta lactamases. This is the first description of an epidemic due to an ESBL-producing member of the family Enterobacteriaceae in Malagasy hospitals. PMID- 19282057 TI - Decolonisation of patients with wounds colonised by MRSA. PMID- 19282058 TI - The power of vivid experience in hand hygiene compliance. AB - In recent years, explicit behavioural theories have been used in some research into hand hygiene behaviour. One of the most prominent of these has been the theory of planned behaviour (TPB). In this qualitative study aimed at increasing understanding of infection prevention practice in the acute care setting, TPB was identified as a suitable framework for the emergence of new insights that have the potential to improve the power of existing education and training. The theory emerging from the research was based on a finding that individual experience is of greater import than formal education in explaining hand hygiene behaviour. This indicated that exposure to vivid vicarious experience is a potential means to improving the power of existing training methods and increasing the propensity for instilling sustainable adequate hand hygiene habits. PMID- 19282059 TI - Don't rush to judge: drawing conclusions about treatment based on observational studies. PMID- 19282060 TI - Patient satisfaction as a function of emergency department previsit expectations. AB - STUDY OBJECTIVES: This study measures the effect of meeting emergency department (ED) patients' expectations for diagnostic and therapeutic interventions on patient satisfaction. METHODS: This was a cross-sectional study of consecutive patients during block enrollment periods surveyed at the beginning and end of their ED visits. On arrival patients or their surrogates were surveyed about the specific interventions they expected during their visit. After completion of ED care, they were surveyed about their level of satisfaction with the entire encounter, assessment of their provider's interpersonal skills, impression of time spent waiting in the ED, and perceived waiting time. Satisfaction was assessed with categorical responses. The degree of concordance of interventions expected and interventions provided was analyzed to determine their effect on overall ED visit satisfaction. RESULTS: Nine hundred eighty-seven patients presented during enrollment periods, 821 met inclusion criteria, and complete data were collected on 504 patient encounters. Twenty-nine percent had no previsit expectations of diagnostic or therapeutic interventions, 24% had a single reported expectation, 47% had multiple intervention expectations. After adjusting for potential confounders, we could not demonstrate a relationship between fulfillment of expectations and satisfaction. We did find a very strong relationship between highly ranked provider interpersonal skills and ED satisfaction (probability ratio of being "very satisfied" 8.6; 95% confidence interval 4.7 to 15.6). Other factors associated with high ED encounter satisfaction were adequate explanations for waiting times and perception of total time in the ED. CONCLUSION: Overall satisfaction was strongly correlated with patient's assessment of the physician's interpersonal skills and was not correlated with whether the physician had met expectations about diagnostic and therapeutic interventions. PMID- 19282061 TI - Prevalence of past year assault among inner-city emergency department patients. AB - STUDY OBJECTIVE: We determine the rates of past year nonpartner violent assault, both victimization and aggression, and assess variables associated with nonpartner violent assault, particularly with regard to substance use. METHODS: A cross-sectional computerized standardized survey study was conducted to assess nonpartner violent assault, physical and mental health, and substance use among patients presenting to an inner-city ED during 2 years. Patients (aged 19 to 60 years) with normal vital signs in an urban emergency department (ED) from 9 am to 11 pm were eligible; pregnant patients and those with a chief complaint of psychiatric evaluation were excluded. Logistic regression analyses were conducted to predict any nonpartner violent assault. RESULTS: Ten thousand seven hundred forty-four patients were enrolled (80% response rate); 14% of the sample reported any past year nonpartner violent assault (9% perpetration; 11% victimization). Findings from regression analyses found that participants with any past year nonpartner violent assault (victimization or aggression) were more likely than their counterparts to be men (2.2), to be single (1.5), to be unemployed (1.1), to present to the ED for injury (1.9), and to report poor physical health (1.3) or poor mental health (1.9). They were less likely to be black (0.8), or older (0.95). Alcohol use (1.7), marijuana use (2.4), cocaine use (3.1), prescription drug use (1.4), and past treatment (1.7) were associated with experiencing past year nonpartner violent assault. CONCLUSION: Fourteen percent of patients seeking care in this inner-city ED experience violence with a nonpartner. Substance use specifically cocaine-was the strongest predictor of any nonpartner violent assault. PMID- 19282062 TI - An analysis of the Association of Society of Chest Pain Centers Accreditation to American College of Cardiology/American Heart Association non-ST-segment elevation myocardial infarction guideline adherence. AB - STUDY OBJECTIVE: Since 2003, the Society of Chest Pain Centers (SCPC) has provided hospital accreditation for acute coronary syndrome care processes. Our objective is to evaluate the association between SCPC accreditation and adherence to the American College of Cardiology/American Heart Association (ACC/AHA) evidence-based guidelines for non-ST-segment elevation myocardial infarction (NSTEMI). The secondary objective is to describe the clinical outcomes and the association with accreditation. METHODS: We conducted a secondary analysis of data from patients with NSTEMI enrolled in the Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes With Early Implementation of the ACC/AHA Guidelines (CRUSADE) quality improvement initiative in 2005. The analysis explored differences between SCPC-accredited and nonaccredited hospitals in evidence-based therapy given within the first 24 hours (including aspirin, beta-blocker, glycoprotein IIb/IIIa inhibitors, heparin, and ECG within 10 minutes). RESULTS: Of 33,238 patients treated at 21 accredited hospitals and 323 nonaccredited hospitals, those at SCPC-accredited centers (n=3,059) were more likely to receive aspirin (98.1% versus 95.8%; odds ratio [OR] 1.73; 95% confidence interval [CI] 1.06 to 2.83) and beta-blockers (93.4% versus 90.6%; OR 1.68; 95% CI 1.04 to 2.70) within 24 hours than patients at non-SCPC-accredited centers (n=30,179). No difference was observed in obtaining a timely ECG (40.4% versus 35.2%; OR 1.28; 95% CI 0.98 to 1.67), administering a glycoprotein IIb/IIIa inhibitor (OR 1.30; 95% CI 0.93 to 1.80), or administering heparin (OR 1.12; 95% CI 0.74 to 1.70). Also, there was no significant difference in risk adjusted mortality for patients treated at SCPC hospitals versus nonaccredited hospitals (3.4% versus 3.5%; adjusted OR 1.17; 95% CI 0.88 to 1.55). CONCLUSION: SCPC-accredited hospitals had higher NSTEMI ACC/AHA evidence-based guideline adherence in the first 24 hours of care on 2 of the 5 measures. No difference in outcomes was observed. Further studies are needed to better understand the association between SCPC accreditation and improved care for patients with acute coronary syndrome. PMID- 19282063 TI - Misdiagnosis of long QT syndrome as epilepsy at first presentation. AB - STUDY OBJECTIVE: Long QT syndrome has significant mortality, which is reduced with appropriate management. It is known that long QT syndrome masquerades as other conditions, including seizure disorders. We aim to evaluate a series of patients with genetically confirmed long QT syndrome to establish the frequency of delayed recognition. We also examine causes and potential consequences of diagnostic delay. METHODS: A consecutive case series of patients with long QT syndrome was identified through the Cardiac Inherited Disease Registry in New Zealand between 2000 and 2005. Detailed retrospective review of 31 cases was undertaken. The primary outcome was the time from first presentation with sudden loss of consciousness to a diagnosis of long QT syndrome. If the diagnosis was not made at the initial presentation, it was considered delayed. For the patients with a delayed diagnosis, the median duration of delay was compared between the subgroup of patients initially misdiagnosed with epilepsy and the others. RESULTS: Genetic mutations in 31 probands were consistent with long QT type 1 in 18 (58%) patients, long QT type 2 in 10 (32%) and long QT type 3 in 3 (10%). Median age at diagnosis was 21 years (1 day to 54 years). Thirteen patients (39%) experienced diagnostic delay after presentation with syncope or seizure: median delay 2.4 years (2 months to 23 years). Electroencephalograms were obtained in 10 patients; 5 were diagnosed with epilepsy. For those labeled epileptic, diagnostic delay was significantly longer than with other misdiagnoses: estimated median difference 9.75 years (95% confidence interval 7.6 to 20.7 years). During the delay period, 4 sudden unexplained deaths reportedly occurred in young relatives. Ten of the 13 had an ECG before diagnosis, with unrecognized pulse rate-corrected QT interval prolongation in 8 cases (range 0.47 to 0.65 seconds). CONCLUSION: Delayed diagnosis of long QT syndrome is frequent. Symptoms are often attributed to alternative diagnoses, most commonly seizure disorder. Patients labeled as epileptic experience a particularly long diagnostic delay. ECGs were frequently requested, but interpretation errors were common. Given the potentially preventable mortality of long QT syndrome, emergency physicians investigating syncope and seizure should maintain a high index of suspicion. PMID- 19282064 TI - Evaluation of an asynchronous physician voicemail sign-out for emergency department admissions. AB - STUDY OBJECTIVE: Communication failures contribute to errors in the transfer of patients from the emergency department (ED) to inpatient medicine units. Oral (synchronous) communication has numerous benefits but is costly and time consuming. Taped (asynchronous) communication may be more reliable and efficient but lacks interaction. We evaluate a new asynchronous physician-physician sign out compared with the traditional synchronous sign-out. METHODS: A voicemail based, semistructured sign-out for routine ED admissions to internal medicine was implemented in October 2007 at an urban, academic medical center. Outcomes were obtained by pre- and postintervention surveys of ED and internal medicine house staff, physician assistants, and hospitalist attending physicians and by examination of access logs and administrative data. Outcome measures included utilization; physician perceptions of ease, accuracy, content, interaction, and errors; and rate of transfers to the ICU from the floor within 24 hours of ED admission. Results were analyzed both quantitatively and qualitatively with standard qualitative analytic techniques. RESULTS: During September to October 2008 (1 year postintervention), voicemails were recorded about 90.3% of medicine admissions; 69.7% of these were accessed at least once by admitting physicians. The median length of each sign-out was 2.6 minutes (interquartile range 1.9 to 3.5). We received 117 of 197 responses (59%) to the preintervention survey and 113 of 206 responses (55%) to the postintervention survey. A total of 73 of 101 (72%) respondents reported dictated sign-out was easier than oral sign-out and 43 of 101 (43%) reported it was more accurate. However, 70 of 101 (69%) reported that interaction among participants was worse. There was no change in the rate of ICU transfer within 24 hours of admission from the ED in April to June 2007 (65/6,147; 1.1%) versus April to June 2008 (70/6,263; 1.1%); difference of 0%, 95% confidence interval -0.4% to 0.3%. The proportion of internists reporting at least 1 perceived adverse event relating to transfer from the ED decreased a nonsignificant 10% after the intervention (95% confidence interval -27% to 6%), from 44% preintervention (32/72) to 34% postintervention (23/67). CONCLUSION: Voicemail sign-out for ED-internal medicine communication was easier than oral sign-out without any change in early ICU transfers or the perception of major adverse events. However, interaction among participants was reduced. Voicemail sign-out may be an efficient means of improving sign-out communication for stable ED admissions. PMID- 19282065 TI - Evidence-based emergency medicine/systematic review abstract. Does this patient have irritable bowel syndrome? PMID- 19282066 TI - Age-related gray matter volume changes in the brain during non-elderly adulthood. AB - Previous magnetic resonance imaging (MRI) studies described consistent age related gray matter (GM) reductions in the fronto-parietal neocortex, insula and cerebellum in elderly subjects, but not as frequently in limbic/paralimbic structures. However, it is unclear whether such features are already present during earlier stages of adulthood, and if age-related GM changes may follow non linear patterns at such age range. This voxel-based morphometry study investigated the relationship between GM volumes and age specifically during non elderly life (18-50 years) in 89 healthy individuals (48 males and 41 females). Voxelwise analyses showed significant (p<0.05, corrected) negative correlations in the right prefrontal cortex and left cerebellum, and positive correlations (indicating lack of GM loss) in the medial temporal region, cingulate gyrus, insula and temporal neocortex. Analyses using ROI masks showed that age-related dorsolateral prefrontal volume decrements followed non-linear patterns, and were less prominent in females compared to males at this age range. These findings further support for the notion of a heterogeneous and asynchronous pattern of age related brain morphometric changes, with region-specific non-linear features. PMID- 19282067 TI - Antiphospholipid syndrome induction exacerbates a transgenic Alzheimer disease model on a female background. AB - The antiphospholipid syndrome (APS) is characterized by antiphospholipid antibodies (aPL) and vascular brain disease which is often associated with dementia. We examined the neurodegenerative pathological processes underlying APS by inducing APS in a transgenic animal model of Alzheimer's disease. Female C57B6/SJL mice carrying the APP(695)SWE mutation (Tg2576) and wild-type (wt) controls were immunized with beta2-glycoprotein-I (APS mice) or adjuvant alone (controls) at 4 months of age. At the age of 8 months the APP-APS mice developed high levels of aPL associated with motor hypoactivity in a staircase test (p<0.03 by t-test) and impaired performance in the cognitive T-maze (p<0.02 for main effect of treatment by repeated measures ANOVA) relative to APP-CFA mice and controls. wt-APS and wt-control did not differ significantly in their behavior or cognition. Histological studies revealed mature plaques only in the APP-APS group which also had higher amyloid load and number of activated microglia compared to all other groups. The results indicate a significant interaction between APP genotype and the induction of APS on a female background. The mechanisms involved may also be important in human APS-AD co-morbidity. PMID- 19282068 TI - [Information needs of patients in primary care from the perspective of health professionals. A Delphi study]. AB - OBJECTIVE: To determine the information needed by patients in relation to the supply of primary care services from the perspective of health professionals. METHODS: The Delphi technique was used as a consensus method. Seventy experts were asked to participate: 35 primary care physicians, 32 nurse practitioners, and three pediatricians in rural and non-rural health centers in the Health Area of Salamanca between May and December 2006. The questionnaires used were structured into six sections: patients' knowledge of their health, services in their health centers, administrative topics, the media, information expectations, and communication barriers. In the third questionnaire, prioritized answers were scored between 5 points and 1 point according to their importance. RESULTS: All three questionnaires were completed by 44 experts (62.8%). The information needs with the highest scores were information on patients' diseases (4.0 points), subjects related to patients' responsibility for their health (3.8) and preventive advice (3.6), followed by subjects relating to access (4.2), emergencies (3.4), physicians' and nurse practitioners' services (3.6), and places to go to get information (4.0). The lack of coordination between primary and hospital care (4.1) and excessive bureaucracy (3.5) were considered to limit communication and information. CONCLUSIONS: The factors identified focus on contents that allow patients to get involved in their own process and to take active part in the decision-making that concerns them. Other important factors were information about services and the access to them. PMID- 19282069 TI - [Impact of immigration on hospital care: utilization, case-mix, and economic effects]. AB - OBJECTIVES: To identify the impact of immigration on health services by comparing hospital discharges, case-mix, and economic effects between immigrants and the native population. METHODS: We performed a retrospective longitudinal study of acute-care hospital admissions in Murcia (Spain) registered in the Minimum Data Set from 2004-2005. The groups to be compared, established on the basis of country of birth, were <>, <> and <>. Diagnoses were codified using the ICE-9-CM and were grouped by means of the All Patient-Diagnosis Related Groups (AP-DRG) version 18. Utilization rates were calculated by the time of medical insurance. Economic effects were calculated through DRG weights for 2004. RESULTS: There were 196,275 discharges, with 2,590,376 person-years of insurance. The frequency of discharges was 75.8 per thousand among Spaniards, 64.3 per thousand among immigrants from Europe-25 and 73.8 per thousand among immigrants from the remaining countries. The most frequent causes of admission were related to pregnancy, childbirth and the puerperium. Cost per admission was 3,529 euro in Spaniards, 3,231 euro in persons from Europe-25 and 2,423 euro in persons from the remaining countries. The average cost per year of insurance was 263 euro for Spaniards, 217 euro for immigrants from Europe-25 and 219 euro for those from the remaining countries. CONCLUSIONS: Hospital utilization and costs per admission and for person-year of insurance are higher in Spaniards than in immigrants, especially the group from <>. In this group, case-mix is also different. PMID- 19282070 TI - [Non necrotizing bacterial cellulitis and bacteriemia due to Shewanella putrefaciens]. AB - Shewanella putrefaciens is a Gram negative opportunistic pathogen which causes skin and soft tissue infections and bacteriemia in immunocompromized patients. We report a 86-year-old man, who presented with an infectious cellulitis of the leg associated with Shewanella putrefaciens bacteriemia. This patient was treated by mycophenolate mofetil for a bullous pemphigoid resistant to corticotherapy. PMID- 19282071 TI - [Magnetic resonance imaging and claustrophobia]. PMID- 19282072 TI - Craniomaxillofacial fibrous dysplasia: a 10-year database 1996-2006. AB - Fibrous dysplasia is a rare bone disease caused by an abnormal proliferation of fibrous tissue in bone. We retrospectively evaluated eight patients (female to male ratio 3:1, mean age 22.5 years, range 10-32) with a monostotic form who were treated between 1996 and 2006. Two each were affected in the lower jaw, the upper jaw, the midface, and the frontoparietal region. Most patients were referred because of a painless swelling. Biopsy specimens from two patients were examined, six patients had modelling osteotomies, two of whom had further operations because of progressive enlargement. There was no visual impairment or malignant transformation. Fibrous dysplasia should be treated as conservatively as possible, but in cases of functional disturbance that results from malignant transformation, or from the involvement of the optic foramen or the foramen magnum, an immediate operation is needed. Disfigurement can be another reason for operation. When there is a risk of malignant transformation, follow-up of patients is recommended. PMID- 19282073 TI - Umbilical cord blood transplantation: pros, cons and beyond. AB - Large body of clinical and scientific data has been generated since the first cord blood transplantation (CBT) was performed in 1989. Superior immune plasticity of CB grafts, that allows for less stringent HLA matching, is especially valuable in the face of a persistently growing need for unrelated donor (UD) transplants. Limited cell dose remains the main setback of CBT, particularly in adult population. New strategies, such as transplantation with two cord blood units or using non-myeloablative conditioning, have remarkably expanded the availability of CB transplants in adults with hematological malignancies. Clinical trials with in vitro expanded CB-derived stem cells are under way. Currently cord blood is considered a second best choice after matched bone marrow. However, results of recent international studies indicate that in particular clinical settings, such as in children with leukemia, CB may become a frontline hematopoietic stem cell (HSC) source for transplantation. Recent advances in understanding the unique biology of cord blood will further expand indications for its use in different settings, including those beyond hematopoietic stem cells transplantation (HSCT). PMID- 19282074 TI - Self-organizing feature map (neural networks) as a tool to select the best indicator of road traffic pollution (soil, leaves or bark of Robinia pseudoacacia L.). AB - Concentrations of the elements Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn were measured in the leaves and bark of Robinia pseudoacacia and the soil in which it grew, in the town of Olesnica (SW Poland) and at a control site. We selected this town because emission from motor vehicles is practically the only source of air pollution, and it seemed interesting to evaluate its influence on soil and plants. The self-organizing feature map (SOFM) yielded distinct groups of soils and R. pseudoacacia leaves and bark, depending on traffic intensity. Only the map classifying bark samples identified an additional group of highly polluted sites along the main highway from Wroclaw to Warszawa. The bark of R. pseudoacacia seems to be a better bioindicator of long-term cumulative traffic pollution in the investigated area, while leaves are good indicators of short-term seasonal accumulation trends. PMID- 19282075 TI - Analysis of phytochelatin complexes in the lead tolerant vetiver grass [Vetiveria zizanioides (L.)] using liquid chromatography and mass spectrometry. AB - Ethylenediamene tetraacetic acid (EDTA) has been used to mobilize soil lead (Pb) and enhance plant uptake for phytoremediation. Chelant bound Pb is considered less toxic compared to free Pb ions and hence might induce less stress on plants. Characterization of possible Pb complexes with phytochelatins (PCn, metal-binding peptides) and EDTA in plant tissues will enhance our understanding of Pb tolerance mechanisms. In a previous study, we showed that vetiver grass (Vetiveria zizanioides L.) can accumulate up to 19,800 and 3350 mg Pb kg(-1) dry weight in root and shoot tissues, respectively; in a hydroponics set-up. Following the basic incubation study, a greenhouse experiment was conducted to elucidate the efficiency of vetiver grass (with or without EDTA) in remediating Pb-contaminated soils from actual residential sites where Pb-based paints were used. The levels of total thiols, PCn, and catalase (an antioxidant enzyme) were measured in vetiver root and shoot following chelant-assisted phytostabilization. In the presence of 15 mM kg (-1) EDTA, vetiver accumulated 4460 and 480 mg Pb kg( 1) dry root and shoot tissue, respectively; that are 15- and 24-fold higher compared to those in untreated controls. Despite higher Pb concentrations in the plant tissues, the amount of total thiols and catalase activity in EDTA treated vetiver tissues was comparable to chelant unamended controls, indicating lowered Pb toxicity by chelation with EDTA. The identification of glutathione (referred as PC1) (m/z 308.2), along with chelated complexes like Pb-EDTA (m/z 498.8) and PC(1)-Pb-EDTA (m/z 805.3) in vetiver root tissue using electrospray tandem mass spectrometry (ES-MS) highlights the possible role of such species towards Pb tolerance in vetiver grass. PMID- 19282076 TI - Association of single-nucleotide polymorphisms in JAK3, STAT4, and STAT6 with new cardiovascular events in incident dialysis patients. AB - BACKGROUND: Increasing evidence supports a role for cell-mediated immunity in the pathogenesis of cardiovascular disease. Single-nucleotide polymorphisms (SNPs) in JAK3, STAT4, and STAT6 of the Janus kinase-signal transducer and activator of transcription (Jak-Stat) signal transduction pathway were examined for association with time to new cardiovascular events in incident dialysis patients from the Choices for Healthy Outcomes in Caring for End-Stage Renal Disease Study. STUDY DESIGN: Prospective cohort study. SETTING & PARTICIPANTS: 764 white (n = 518) and black (n = 246) participants from 79 dialysis centers. PREDICTOR: SNPs in JAK3, STAT4, and STAT6 selected using a pairwise approach to identify a maximally informative set of tag SNPs for populations of European and African descent. OUTCOMES & MEASUREMENTS: Cox proportional hazards models were used to estimate unadjusted and multivariable-adjusted hazard ratios (HRs) for incident cardiovascular disease events after dialysis therapy initiation associated with each race-specific SNP. RESULTS: 2 European tag SNPs (rs3212780 and rs3213409) in JAK3 were associated with new cardiovascular disease events in white patients with unadjusted HRs of 1.92 (P < 0.001) and 1.82 (P = 0.07), respectively. One dual-tag SNP (rs3212752) in JAK3 was associated with new cardiovascular events in white patients with an unadjusted HR of 2.09 (P < 0.001) and in black patients with an HR of 2.07 (P = 0.007). SNP rs3213409 codes for a valine to isoleucine change at amino acid 722, a potentially functional mutation. SNPs in STAT4 and STAT6 were not associated with cardiovascular events after the initiation of dialysis therapy. LIMITATIONS: This study does not provide direct evidence for the mechanism of increased risk. Replication in independent cohorts is necessary. CONCLUSIONS: Genetic polymorphisms in the Jak-Stat signaling pathway are associated with an increased risk of new cardiovascular events in incident dialysis patients. PMID- 19282077 TI - Partner- and partnership-related risk factors for preterm birth among low-income women in Lima, Peru. AB - A woman's partner and the characteristics of their partnership can play an important role in the health of her pregnancy. Yet, with the notable exception of intimate partner violence, there has been little previous research addressing the associations between partner- or partnership-related factors and birth outcomes. This analysis tested the hypothesis that risk factors related specifically to partner or partnership characteristics increased the risk for preterm birth. Between 2003 and 2005, a total of 580 preterm cases (20-36 weeks gestational age at delivery) and 633 term controls (> or =37 weeks) were selected from women delivering at an obstetric hospital in Lima, Peru. Each woman completed a confidential, structured interview and provided biological specimens within 48 h after delivery. Multivariable logistic regression was used to assess associations between partner and partnership characteristics and preterm birth. After adjustment for behavioral, demographic, and obstetric risk factors, ever having had a partner with a history of drug use (aOR = 1.91, 95% CI 1.22-2.99), ever having had anal sex (aOR = 1.40, 95% CI 1.07-1.84), having a current partner with a history of visiting prostitutes (aOR = 1.69, 95% CI 1.22-2.33), and perceiving one's current partner as a "womanizer" (aOR = 1.34, 95% CI 1.02-1.77) were significantly associated with an elevated risk of preterm birth when tested in separate models. These four factors were then used to create a composite partnership risk score, which showed an increasing dose-response relationship with preterm birth risk (per additional partner risk factor: aOR = 1.31, 95% CI 1.16-1.49). These results highlight the importance of considering a broader set of risk factors for preterm birth, specifically those related to a woman's partner and partnership characteristics. Further research could clarify the specific mechanisms through which these partner and partnership characteristics may increase the risk of preterm birth. PMID- 19282078 TI - Tobacco control policies are egalitarian: a vulnerabilities perspective on clean indoor air laws, cigarette prices, and tobacco use disparities. AB - This study models independent associations of state or local strong clean indoor air laws and cigarette prices with current smoker status and consumption in a multilevel framework, including interactions with educational attainment, household income and race/ethnicity and the relationships of these policies to vulnerabilities in smoking behavior. Cross sectional survey data are employed from the February 2002 panel of the Tobacco Use Supplement of the Current Population Survey (54,024 individuals representing the US population aged 15-80). Non-linear relationships between both outcome variables and the predictors were modeled. Independent associations of strong clean indoor air laws were found for current smoker status (OR 0.66), and consumption among current smokers (-2.36 cigarettes/day). Cigarette price was found to have independent associations with both outcomes, an effect that saturated at higher prices. The odds ratio for smoking for the highest versus lowest price over the range where there was a price effect was 0.83. Average consumption declined (-1.16 cigarettes/day) over the range of effect of price on consumption. Neither policy varied in its effect by educational attainment, or household income. The association of cigarette price with reduced smoking participation and consumption was not found to vary with race/ethnicity. Population vulnerability in consumption appears to be structured by non-white race categories, but not at the state and county levels at which the policies we studied were enacted. Clean indoor air laws and price increases appear to benefit all socio-economic and race/ethnic groups in our study equally in terms of reducing smoking participation and consumption. PMID- 19282079 TI - Combining education and video-based contact to reduce stigma of mental illness: "The Same or Not the Same" anti-stigma program for secondary schools in Hong Kong. AB - This study examined the effects of three versions of school-based stigma reduction programs against mental illness - education, education followed by video-based contact (education-video), and video-based contact followed by education (video-education). The participants, 255 students from three secondary schools in Hong Kong, completed measures of stigmatizing attitudes (Public Stigma Scale), social distance (Social Distance Scale), and knowledge about schizophrenia (Knowledge Test) at pre-test, post-test, and 1-month follow-up. Results suggested that adding video-based contact to education could significantly improve program effectiveness only when video-based contact was presented after but not prior to education. In comparison with the education condition, the education-video condition showed larger improvements in stigmatizing attitudes at post-test, in social distance at both post-test and follow-up, and in knowledge at follow-up. However, such differences were not observed when the education condition was compared with the video-education condition. Implications of these findings for future research are discussed. PMID- 19282080 TI - Health interventions and the persistence of rumour: the circulation of sterility stories in African public health campaigns. AB - Public health programmes have done enormous good in Africa and elsewhere in the global south, but have also been met with skepticism. This skepticism often takes the form of rumours about the motives or the results of the public health intervention. One recurrent theme in such rumours is the centrality of reproductive bodies (both male and female), and the perception that these bodies are being rendered sterile by toxic compounds given under the guise of improving health. Public health operations research has identified these rumours as significant obstacles to programme delivery, but they have been treated primarily as failures in communication, to be rectified by the provision of more accurate information. Using reports of such rumours from public health interventions in Africa, with emphasis on vaccines, I argue that these rumours are more than simply stories which are not true. The widespread rumour of sterility is a way of articulating broadly shared understandings about reproductive bodies, collective survival, and global asymmetries of power. I use Foucault's notion of biopolitics to theorize international public health programmes, and introduce the concept of counter-epistemic convergence to account for the ubiquity and persistence of sterility rumours. PMID- 19282081 TI - Experiencing and regulating sadness: physiological and cognitive effects. AB - No prior study has examined the two most prominent response-focused regulation strategies (suppression and exaggeration) using a within-subjects design. Utilizing this design allows for a direct comparison of physiological patterns and cognitive impairment associated with such efforts. One hundred and nine participants were asked to view a series of three films, each preceded by a 10 second instructional slide which indicated the regulation strategy they were to perform (natural-watch, exaggerate, or suppress). Exaggeration was associated with increased sympathetic activation as indicated by an increase in galvanic skin conductance level (GSL) and shortened pre-ejection period (PEP). Suppression, much like the natural-watch condition, was associated with greater sympathetic withdrawal (i.e., decreased GSL, longer PEP). Both suppression and exaggeration led to reduced memory for the emotional movie, with exaggeration causing more impairment than suppression. Results suggest that exaggeration and suppression not only have very different behavioral manifestations, but physiological outcomes as well when utilized during a sad context. PMID- 19282082 TI - Timing dysfunctions in schizophrenia span from millisecond to several-second durations. AB - Schizophrenia may be associated with a fundamental disturbance in the temporal coordination of information processing in the brain, leading to classic symptoms of schizophrenia such as thought disorder and disorganized and contextually inappropriate behavior. However, the majority of studies that have examined timing behavior in schizophrenia have employed temporal durations in the range of several seconds, which requires higher cognitive processes beyond initial sensory registration for temporal encoding. Accordingly, the present study assessed both millisecond and several-second duration estimates in schizophrenia using a well established task of time perception. Twenty-eight individuals with schizophrenia and 31 non-psychiatric control participants completed two temporal bisection tasks, which required participants to make temporal judgments about auditory durations ranging from either 300 to 600 ms or 3000 to 6000 ms. Participants with schizophrenia displayed significantly greater timing variability under both millisecond and several-second timing conditions than the non-psychiatric group. These findings were consistent with parameter estimates obtained from a quantitative model of time estimation, and provide evidence for a fundamental timing deficit in schizophrenia that may be independent of the length of the to be-timed duration. PMID- 19282083 TI - The effect of preparation designs on the marginal and internal gaps in Cerec3 partial ceramic crowns. AB - OBJECTIVES: The purpose of this study was to evaluate the marginal and internal gaps in Cerec3 partial ceramic crowns (PCCs) of three different preparation designs in vitro using microcomputed tomography (microCT). METHODS: Cerec3 PCCs of three different preparation designs (n=20) were fabricated according to the following: Group I-conventional functional cusp capping/shoulder preparation, Group II-horizontal reduction of cusps and Group III-complete reduction of cusps/shoulder preparation. After fixation of PCCs, the microCT scanning was performed. For obtaining the average internal gap (AIG), the microCT sections were reconstructed 3-dimensionally, and then the total volume of the internal gap was divided by the contact surface area. The 2-dimensional (2D) microCT views were used to investigate the gaps at predetermined key positions in seven bucco lingual sections and three mesio-distal cross sections. The gaps were measured using the microCT at each reference point. Statistical analysis was performed using one-way ANOVA and Tukey's test. RESULTS: For the 3D reconstruction technique, the AIGs were as followed: Group I 197.3+/-48.2microm, Group II 171.2+/-45.1microm, and Group III 152.7+/-27.1microm. For the 2D microCT views, the gaps of each group were the smallest on the margins ranging from 35.4+/-32.2 to 128.4+/-69.5microm, and the largest on the horizontal or angle walls ranging from 184.5+/-41.2 to 406.5+/-176.1microm. According to the results, Group I showed larger marginal and internal gaps compared with the other groups. CONCLUSIONS: For the PCCs, the simplified designs (Groups II and III) demonstrated superior results compared to the traditional cusp capping design (Group I). The marginal gaps were smaller than the internal gaps in all groups. PMID- 19282084 TI - Life after survival: long-term daily functioning and quality of life after an out of-hospital cardiac arrest. AB - BACKGROUND: Information about long-term consequences of cardiac arrest is sparse. Because the survival rate is expected to increase, better knowledge of long-term functioning and quality of survival is essential. OBJECTIVES: To determine the level of functioning of out-of-hospital cardiac arrest survivors 1-6 years later, and to evaluate the predictive value of medical variables on long-term functioning. METHODS: A retrospective cohort study including 63 survivors of an out-of-hospital cardiac arrest, admitted to a Dutch University hospital between 2001 and 2006. Participants received a questionnaire by post. Primary outcome measures were: participation in society (Community Integration Questionnaire) and quality of life (SF-36). Secondary outcome measures were: physical, cognitive and emotional impairment, daily functioning and caregiver strain. Statistical analyses included multiple regression analyses. RESULTS: On average 3 years post cardiac arrest, 74% of the patients experienced a low participation level in society compared with the general population. Over 50% reported severe fatigue, 38% feelings of anxiety and/or depression and 24% a decreased quality of life. Caregivers reported stress related responses, feelings of anxiety and lower quality of life. Seventeen percent of the caregivers reported high caregiver strain, which was associated with the patient's level of functioning. Gender, age, percutaneous coronary intervention (PCI) and therapeutic hypothermia contributed to outcome on at least one domain of long-term functioning. CONCLUSIONS: After surviving an out-of-hospital cardiac arrest, many patients and partners encounter extensive impairments in their level of functioning and quality of life. Gender, age, PCI and therapeutic hypothermia are associated with differences in long-term functioning of patients. PMID- 19282085 TI - Intravenous fat emulsion therapy for intentional sustained-release verapamil overdose. AB - We report the first case of sustained-release verapamil toxicity treated with Intralipid fat emulsion (IFE). Toxicity was confirmed by elevated serial serum verapamil and metabolite, norverapamil, levels. Most previously reported cases of IFE therapy involve local anaesthetic toxicity and cardiac arrest. Our patient was in shock despite standard therapy. No adverse events were noted and the patient fully recovered. PMID- 19282086 TI - Temporal and spatial variations in spontaneous Ca events and mechanical activity in pregnant rat myometrium. AB - OBJECTIVES: The aim of this study was to investigate the temporal and spatial characteristics of spontaneous Ca signals in pregnant rat myometrium. STUDY DESIGN: Confocal imaging of longitudinal strips of 21-day pregnant rats loaded with the Ca sensitive indicator Fluo-4, was combined with measurements of mechanical activity in uterine smooth muscle cells, in situ and freshly isolated. RESULTS: Our results show that the Ca transients in pregnant uterine tissue are composed of Ca spikes, which are associated with the spike-like action potentials. There is large variation in the pattern of spontaneous activity in myometrium, ranging from non-propagating Ca spikes confined to individual smooth muscle cells, through to regional and global propagating Ca spikes. Irrespective of the pattern of activity displayed, the Ca signals were always in the form of Ca spikes, singularly or in bursts. These Ca spikes did not show fixed initiations sites, propagated in longitudinal and transverse directions from the initiation regions, and had a variable pattern of propagation in preparations which were not synchronously active. In preparations which showed synchronous activity, Ca spikes singularly or bursts propagated mainly in the transverse direction from the initiation regions. The amplitude of force generated by single spikes was dependent on the number of bundles recruited by the propagating Ca spike within the strip, and was about 30-40% of the maximal force produced by carbachol or high-K stimulation. If Ca spikes appeared in the form of bursts they generated longer lasting fused contractions, the amplitudes of which were dependent on the number and the frequency of Ca spikes in the burst. CONCLUSIONS: Longitudinal myometrium from pregnant rats generates spontaneous Ca spikes which vary in their initiation sites, spatial spread and frequency and are associated with the spike-like action potentials. They are sensitive to the L-type Ca channel blocker, nifedipine. Contractile activity was dependent on the spatial spread of individual Ca spikes and when fully synchronized, produced single submaximal phasic contraction. The number and frequency of bursts of Ca spikes controlled the amplitude and duration of contraction. PMID- 19282087 TI - Extrarenal Wilms' tumor of the uterus with ovarian dermoid cyst. PMID- 19282088 TI - Reproductive Bioengineering 2008. Introduction. PMID- 19282089 TI - Phase-plot analysis of the oxytocin effect on human myometrial contractility. AB - OBJECTIVE: Analysis of uterine contractility in vitro is usually confined to measuring a few traditional parameters of uterine contractility, such as contraction amplitude, frequency and area under the curve. In this paper, we describe parameters that provide additional information obtained from the traces of force and its first derivative. We propose an improved contractility index which is less dependent on variability between samples. STUDY DESIGN: Standard organ bath recording of myometrial contractions in the presence or absence of oxytocin on samples of human myometrium obtained from 26 patients at Caesarean section. The parameters were obtained from the plots of first derivative vs. contractile force (phase portrait plot). RESULTS AND CONCLUSIONS: Oxytocin (1nM) significantly increased the contraction amplitude (Fmax), area under the curve, maximum rate of contraction (CVmax), decreased the maximum rate of relaxation (RVmax) and had no statistically significant effect on the duration of contraction (measured as full width at half amplitude, W50). In addition to the above effects, 10nM oxytocin increased the contraction duration (P=0.0036, n=24). The fraction of force developed at the time of CVmax showed no change at any concentration of oxytocin, while the fraction of force remaining at RVmax was decreased in a dose dependent manner. The least variable (i.e. showing lowest P values in paired Student's t-Test) parameters were the Fmax and CVmax/RVmax. When non-paired t-Test was applied, P value of the CVmax/RVmax remained low, while the variability of Fmax increased reflecting the sample-to-sample variations. The product of the Fmax and CVmax/RVmax, which we propose as uterine contractility index (CI) showed low P values in both paired and non-paired t-Tests. We conclude that the phase plot analysis provides useful additional information on contraction/relaxation properties of human myometrium and the CI is suitable for characterising the contractility of uterine samples with different connective tissue content. PMID- 19282090 TI - Uterine contraction signals--application of the linear synchronization measures. AB - OBJECTIVE: In physiological research, there are not too many studies on multivariate data sets, containing two or more simultaneously recorded time series. It is important to examine synchronization in these kinds of signals. The aim of this study is to present the linear measures: the cross-correlation function, the coherence function, the wavelet cross-correlation and the wavelet coherence to assess synchronization between contractions in different topographic regions of the uterus. STUDY DESIGN: Spontaneous uterine activity was recorded directly by a dual micro-tip catheter (Millar Instruments, Inc., USA). The device consisted of two ultra-miniature pressure sensors. One sensor was placed in the fundus, the other one in the cervix. For this analysis, a healthy patient with normal contractions, a patient with dysmenorrhea, a patient with fibromyomas in the follicular phase, and the patient with endometriosis were selected. RESULTS: For each method the values of synchronization parameters for normal contractions were higher than the values of these parameters for other pairs of signals. The differences between these four groups of the uterine contraction signals were clear. The lowest values of the synchronization measures were in the case of dysmenorrheic patient. CONCLUSION: The analysis of synchronization of the uterine contractions signals may have a diagnostic value. For intrauterine pressure signals results obtained by means of different synchronization methods are different, but consistent. PMID- 19282091 TI - Relation between mechanical properties and microstructure of human fetal membranes: an attempt towards a quantitative analysis. AB - OBJECTIVE: We sought to measure the mechanical baseline behavior of fetal membranes in order to determine constitutive mechanical model parameters for fetal membranes, and to examine their relation to molecular correlates for mechanical function, i.e. collagen and elastin. STUDY DESIGN: The uniaxial stress strain response of nine human term fetal membranes was measured. Methods of nonlinear continuum mechanics were applied for the analysis of the stress-strain curves. Thickness of amnion and chorion were determined from histologic sections for each fetal membrane sample. Complementary biochemical analysis was performed to quantify the soluble collagen and soluble elastin components for each sample. RESULTS: We report a straightforward histologic modality for measurements of amnion and chorion thickness. Average thickness of the amnion and chorion layers were 111+/-78 microm, and 431+/-113 microm, respectively, which are about twice larger than previously reported. The average content of acid-soluble elastin was 2.1% of wet weight and the one of pepsin/acetic acid-soluble collagen was 10.5% of dry weight. Our data show an inverse proportionality between soluble elastin and soluble collagen content. The low strain elastic modulus ranged between 10 and 25 kPa. Correlations were found between biochemical data and mechanical parameters: there is clearly a direct proportionality between small strain elastic modulus and elastin content. Further, a (less pronounced) direct correlation was observed also between soluble collagen content and the parameter governing the increase in stiffness at larger strains in the nonlinear mechanical model. The mechanical tests revealed a relatively low variability for samples from the same membrane but a large variation between donors. The proposed nonlinear model provides a good fit of the experimental data, with a coefficient of determination, R(2), typically in the range of 0.94. Membranes failure originated at the clamping points thus impairing the quantification of ultimate stress and strain. Thus, no correlation was found between maximum stress and collagen or elastin content. CONCLUSIONS: This study provides a starting point for comprehensive quantitative analysis of the relationship between fetal membranes microstructure and their nonlinear deformation behavior. These insights could become useful in identifying potential medical interventions to prevent membranes rupture. PMID- 19282092 TI - Molecular modeling, dynamics and docking studies of purine nucleoside phosphorylase from Streptococcus pyogenes. AB - Purine Nucleoside Phosphorylase (PNP) catalyzes the reversible phosphorolysis of N-glycosidic bonds of purine nucleosides and deoxynucleosides, except for adenosine, to generate ribose 1-phosphate and the purine base. PNP has been submitted to intensive structural studies. This work describes for the first time a structural model of PNP from Streptococcus pyogenes (SpPNP). We modeled the complexes of SpPNP with six different ligands in order to determine the structural basis for specificity of these ligands against SpPNP. Molecular dynamics (MD) simulations were performed in order to evaluate the overall stability of SpPNP model. The analysis of the MD simulation was assessed mainly by principal component analysis (PCA) to explore the trimeric structure behavior. Structural comparison, between SpPNP and human PNP, was able to identify the main features responsible for differences in ligand-binding affinities, such as mutation in the purine-binding site and in the second phosphate-binding site. The PCA analysis suggests a different behavior for each subunit in the trimer structure. PMID- 19282093 TI - Increasing exfiltration from pervious concrete and temperature monitoring. AB - Pervious concrete typically has an infiltration rate far exceeding any expectation of precipitation rate. The limiting factor of a retention based pervious concrete system is often defined by how quickly the underlying soil subgrade will infiltrate the water temporarily stored within the concrete and/or aggregate base. This issue is of particular importance when placing a pervious concrete system on compacted fine textured soils. This research describes the exfiltration from twelve pervious concrete plots constructed on a compacted clay soil in eastern Tennessee, USA. Several types of treatments were applied to the clay soil prior to placement of the stone aggregate base and pervious concrete in an attempt to increase the exfiltration rate, including: 1) control--no treatment; 2) trenched--soil trenched and backfilled with stone aggregate; 3) ripped--soil ripped with a subsoiler; and 4) boreholes--placement of shallow boreholes backfilled with sand. The average exfiltration rates were 0.8 cm d(-1) (control), 4.6 cm d(-1) (borehole), 10.0 cm d(-1) (ripped), and 25.8 cm d(-1) (trenched). The trenched treatment exfiltrated fastest, followed by the ripped and then the borehole treatments, although the ripped and borehole treatments were not different from one another at the 5% level of significance. The internal temperature of the pervious concrete and aggregate base was monitored throughout the winter of 2006-2007. Although the temperature of the pervious concrete dropped below freezing 24 times, freezing concrete temperatures never coincided with free water being present in the large pervious concrete pores. The coldest recorded air temperature was -9.9 degrees C, and the corresponding coldest recorded pervious concrete temperature was -7.1 degrees C. The temperature of the pervious concrete lagged diurnal air temperature changes and was generally buffered in amplitude, particularly when free water was present since the addition of water increases the thermal capacity of the pervious concrete greatly. The temperature of the aggregate base was further buffered to diurnal changes, and no freezing temperatures were recorded. PMID- 19282094 TI - Performance evaluation of methods for two-dimensional displacement and strain estimation using ultrasound radio frequency data. AB - In elastography, several methods for 2-D strain imaging have been introduced, based on both raw frequency (RF) data and speckle-tracking. Although the precision and lesion detectability of axial strain imaging in terms of elastographic signal-to-noise ratio (SNRe) and elastographic contrast-to-noise ratio (CNRe) have been reported extensively, analysis of lateral precision is still lacking. In this paper, the performance of different 2-D correlation RF- and envelope-based strain estimation methods was evaluated using simulation data and phantom experiments. Besides window size and interpolation methods for subsample displacement estimation, the influence of recorrelation techniques was examined. Precision and contrast of the measured displacements and strains were assessed using the difference between modeled and measured displacements, SNRe and CNRe. In general, a 2-D coarse-to-fine displacement estimation method is favored, using envelope data for window sizes exceeding the theoretical upper bound for strain estimation. Using 2-D windows of RF data resulted in better displacement estimates for both the axial and lateral direction than 1-D RF-based or envelope-based techniques. Obtaining subsample lateral displacement estimates by fitting a predefined shape through the cross-correlation function (CCF) yielded results similar to those obtained with up-sampling of RF data in the lateral direction. Using a CCF model was favored because of the decreased computation time. Local aligning and stretching of the windows (recorrelation) resulted in an increase of 2-17 and 6-7 dB in SNRe for axial and lateral strain estimates, respectively, over a range of strains (0.5 to 5.0%). For a simulated inhomogeneous phantom (2.0% applied strain), the measured axial and lateral SNRes were 29.2 and 20.2 dB, whereas the CNRes were 50.2 dB and 31.5 dB, respectively. For the experimental data, lower SNRe (axial: 28.5 dB; lateral: 17.5 dB) and CNRe (axial: 39.3 dB; lateral: 31 dB) were found. In conclusion, a coarse-to-fine approach is favored using RF data on a fine scale. The use of 2D parabolic interpolation is favored to obtain subsample displacement estimates. Recorrelation techniques, such as local aligning and stretching, increase SNRe and CNRe in both directions. PMID- 19282096 TI - Enhancement of microbubble mediated gene delivery by simultaneous exposure to ultrasonic and magnetic fields. AB - It has been shown in previous studies that gene delivery can be enhanced by a variety of minimally-invasive techniques including: (1) exposure of cells to ultrasound in the presence of DNA and gas microbubbles and (2) exposure of cells to a magnetic field in the presence of DNA conjugated to magnetic nanoparticles. The aim of this work was to investigate whether it was possible to combine the advantages of both these techniques. It was found that transfection of Chinese hamster ovary cells by naked plasmid DNA was enhanced by combined exposure of the cells to ultrasound (10 s at 1 kHz pulse repetition frequency with 40 cycle 1 MHz sinusoidal pulses, 1 MPa peak to peak pressure) and a magnetic field (provided by five square cross-section N52 grade NdFeB magnets 25 x 10 x 10 mm with transversal magnetisation Br = 1.50 T arranged in a Halbach array), in the presence of one of two different microbubble/nanoparticle preparations. The first preparation consisted of phospholipid coated microbubbles mixed with micelles containing magnetic nanoparticles. The second consisted of microbubbles which were themselves magnetically active. These preparations were found to be more effective than either magnetic micelles or phospholipid coated microbubbles alone by a factor of 2.8 (total flux approximately 4 versus 1.4 x 10(6) photon/s) and the results were found to be statistically significant (p < 0.01). Two mechanisms are proposed to explain these observations: firstly, that the magnetic field facilitates close proximity between the cells and the microbubbles and hence increases the likelihood of transfection; second, that there is sensitisation of the cells, as a result of exposure to the magnetic field in the presence of the micelles, which increases their ability to be transfected upon exposure to ultrasound. Further work is in progress to determine which of these mechanisms is the most significant and the potential for other therapeutic applications. PMID- 19282097 TI - Serum proteomic profiling in patients with bladder cancer. AB - BACKGROUND: Despite continuing research for accurate bladder cancer biomarkers, the analytes suffer from lack of sensitivity and specificity. OBJECTIVE: To search for discriminating protein patterns in serum, we used magnetic bead-based separation followed by matrix-assisted laser desorption ionization (MALDI) time of-flight (TOF) mass spectrometry (MS) to identify patients with bladder cancer. DESIGN, SETTING AND PARTICIPANTS: In total, serum samples from 105 patients with bladder cancer, 98 healthy controls, and 45 prostate cancer patients were included in this study. MEASUREMENTS: Serum samples were fractionated by means of surface-activated magnetic beads and were subsequently analyzed with MALDI-TOF MS. Multidimensional data analysis was done to generate algorithms capable of distinguishing between cancer patients and healthy individuals. The algorithms were trained using a training set of 41 bladder cancer patients and 39 healthy controls and were validated with an independent test set of 64 bladder cancer patients and 59 healthy controls. Additionally, 45 prostate cancer samples were used as a third test set. RESULTS AND LIMITATIONS: In the training set, patients with bladder cancer could be identified with an overall sensitivity of 94.1% and specificity of 89.2%. Similar results could be achieved for the test set, showing 96.4% sensitivity and 86.5% specificity. Even the presence of low-stage tumors could be predicted with 96% sensitivity and could be distinguished from higher stage or grade tumors with a sensitivity of 77.3%. Distinction between other tumor stages, however, resulted in lower sensitivity values. CONCLUSIONS: These findings demonstrate that screening for serum protein patterns using MALDI-TOF MS shows high sensitivity and specificity in identifying patients with bladder cancer, regardless of tumor stage. Due to high-throughput capability, the identified differential protein panel may improve the detection of bladder cancer. PMID- 19282098 TI - A randomized phase 1 study of testosterone replacement for patients with low-risk castration-resistant prostate cancer. AB - BACKGROUND: Even in castration-resistant prostate cancer (CRPC), the androgen pathway remains biologically relevant. In preclinical models, androgen therapy for CRPC leads to growth arrest, apoptosis, and tumor shrinkage. OBJECTIVE: This study sought to determine the toxicity and feasibility of a testosterone therapy in early CRPC. DESIGN, SETTING, AND PARTICIPANTS: Prostate cancer patients with progressive disease following androgen ablation, antiandrogen therapy, and withdrawal and no to minimal metastatic disease who were followed at the University of Chicago were randomized to treatment with three doses of transdermal testosterone. INTERVENTION: Patients were treated with transdermal testosterone at 2.5, 5.0, or 7.5 mg/day. MEASUREMENTS: Toxicity, prostate specific antigen (PSA), imaging, quality of life (QoL), and strength were monitored. Treatment was discontinued for significant toxicity, clinical progression, or a 3-fold increase in PSA. RESULTS AND LIMITATIONS: Fifteen men with a median age of 73 yr (range: 62-92) and a median PSA of 11.1 ng/ml (range: 5.2-63.6) were treated. Testosterone increased from castrate to median concentrations of 305 ng/dl, 308 ng/dl, and 297 ng/dl for dosages of 2.5 mg/day (n=4), 5.0 mg/day (n=5), and 7.5 mg/day (n=5), respectively. One patient was taken off of the study at 53 wk due to grade 4 cardiac toxicity. There were no other grade 3 or 4 toxicities related to the study medication, and the grade 2 toxicities were minimal. Only one patient experienced symptomatic progression, and three (20%) patients demonstrated a decrease in PSA (largest was 43%). Median time to progression was 9 wk (range: 2-96), with no detectable difference in the three dose cohorts. There was no significant improvement in QoL, and there was a borderline statistically significant improvement in hand-grip strength with treatment. The study was limited by sample size, single arm, and variability of baseline patient characteristics. CONCLUSIONS: Testosterone is a feasible and reasonably well-tolerated therapy for men with early CRPC. A larger, randomized trial is under way to further characterize efficacy and impact on QoL measures. PMID- 19282099 TI - Editorial comment on: Surgical treatment of 31 complex traumatic posterior urethral strictures associated with urethrorectal fistulas. PMID- 19282100 TI - Surgical treatment of 31 complex traumatic posterior urethral strictures associated with urethrorectal fistulas. AB - BACKGROUND: Urethrorectal fistulas (URF) in patients with complex posterior urethral strictures are rare and difficult to repair surgically. There is no widely accepted standard approach described in the published literature. OBJECTIVE: The aim of this study was to describe the outcomes of various operative approaches for the repair of URFs in patients with complex posterior urethral strictures. DESIGN, SETTING, AND PARTICIPANTS: From January 1985 to December 2007, 31 patients (age: 6-61 yr; mean: 28.4) with URFs secondary to posterior urethral strictures were treated using a perineal or combined abdominal transpubic-perineal approach. INTERVENTIONS: A simple perineal approach was used in 4 patients; a transperineal inferior pubectomy approach was used in 18 patients; and a combined transpubic-perineal approach was used in 9 patients. A bulbospongiosus muscle and subcutaneous dartos pedicle flaps were interposed between the repaired rectum and urethra in 22 patients. The combined transpubic perineal approach used either a gracilis muscle flap (one patient) or a rectus muscle flap (eight patients). MEASUREMENTS: Suprapubic catheterisation was used for bladder drainage, and a urethral silicone stent was left indwelling for 4 wk. RESULTS AND LIMITATIONS: One-stage repair was successful in 4 patients (100%) using the perineal approach, in 16 of 18 patients (88.9%) using the transperineal inferior pubectomy approach, and in 7 of 9 patients (77.8%) using the transpubic perineal approach. Recurrent urethral strictures developed in two cases; one patient required regular dilation, and the other patient was treated successfully with tubed perineoscrotal flap urethroplasty. Recurrent URFs developed in two additional patients. CONCLUSIONS: Surgical approaches for the treatment of URFs associated with complex urethral strictures should be based on a number of considerations including the location of the URF, its aetiology, the length of the urethral strictures, and a history of previous unsuccessful repairs. These results demonstrate that the transperineal-inferior pubic approach may be appropriate as a first-line procedure. PMID- 19282101 TI - Editorial comment on: Surgical treatment of 31 complex traumatic posterior urethral strictures associated with urethrorectal fistulas. PMID- 19282102 TI - Editorial comment on: Serum proteomic profiling in patients with bladder cancer. PMID- 19282103 TI - Editorial comment on: A randomized phase 1 study of testosterone replacement for patients with low-risk castration-resistant prostate cancer. PMID- 19282104 TI - Lack of KRAS and BRAF mutation in renal cell carcinoma. PMID- 19282105 TI - Visible light absorption ability and photocatalytic oxidation activity of various interstitial N-doped TiO2 prepared from different nitrogen dopants. AB - Nitrogen-doped TiO(2) was developed to enable photocatalytic reactions using the visible range of the solar spectrum. This work reports on the synthesis, characterisation and kinetic study of interstitial N-doped TiO(2) prepared by the sol-gel method using three different types of nitrogen dopants: diethanolamine, triethylamine and urea. X-ray diffraction, scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy and UV-visible spectroscopy were used to analyse the titania. Different interstitial N-doped TiO(2) properties, such as absorption ability in the UV-visible light region, redshift in adsorption edge, good crystallisation and composition ratio of titania structures (anatase and rutile) could be obtained from different nitrogen dopants. Amongst investigated nitrogen precursors, diethanolamine provided the highest visible light absorption ability of interstitial N-doped TiO(2) with the smallest energy bandgap and the smallest anatase crystal size, resulting in the highest efficiency in 2-chlorophenol degradation. The photocatalytic activity of all N-doped TiO(2) can be arranged in the following order: TiO(2)/diethanolamine>TiO(2)/triethylamine>TiO(2)/urea>un-doped TiO(2). The initial rate of 2-chlorophenol degradation using the interstitial N-doped TiO(2) with diethanolamine was 0.59 mg/L-min and the kinetic constant was 2.34 x 10( 2)min(-1) with a half-life of 98 min. In all cases, hydroquinone was detected as a major intermediate in the degradation of 2-chlorophenol. PMID- 19282106 TI - Phenol sorption on surfactant-modified Mexican zeolitic-rich tuff in batch and continuous systems. AB - Surfactant modified clinoptilolite-rich tuff was used for the removal of phenol from aqueous solutions. The zeolitic rock from Oaxaca (Mexico) was treated with sodium chloride and then modified with hexadecyltrimethylammonium chloride or bencylcetildimethylammonium chloride in different experimental conditions. Phenol sorption isotherms and column experiments were performed; in both cases, phenol was determined in the aqueous solutions by UV-vis spectroscopy. The results showed that the sorption of phenol depends on the quantities of surfactant in the zeolitic rocks, the kind of surfactant, modification conditions and pH of the solutions. When the initial concentration of phenol increased, the adsorption of phenol in the surfactant-modified zeolite increased and the experimental data was best adjusted to the Langmuir model. The saturation of the columns rapidly reached high percentages. PMID- 19282107 TI - An investigation on the distribution of eight hazardous heavy metals in the suburban farmland of China. AB - Understanding spatial variability of hazardous soil heavy metals is an important precondition for suitably monitoring and evaluating eco-environment quality in the primary agricultural production zone. One hundred topsoils were sampled from the urban-rural transition zone in Taihang Piedmont Plain, China. The contents of eight heavy metals Cu, Zn, Cr, Ni, Pb, Cd, Hg and As were tested for each soil sample, and their spatial patterns were analyzed by the semivariogram approach of geostatistics and geographical information system (GIS) technology. Results showed that Cd concentration exceeded its background level. The local pollution from Cd attributed to the anthropogenic influence. The concentrations of eight hazardous heavy metals are relatively lower than the critical values of the national soil quality standard. The correlation distance of soil heavy metals ranged from 3.28 to 11.63 km, with the eight heavy metals having moderate spatial dependence. Cu, Cr, Ni, Pb and As are associated with and controlled by parent material. The results are helpful for improving agricultural and forest ecosystem in the arid and semiarid region. PMID- 19282108 TI - An evaluation of the efficacy of compound alpha and triclabendazole against two isolates of Fasciola hepatica. AB - Seventy indoor-reared sheep were divided into 10 groups to test the efficacy of the experimental fasciolicide, compound alpha (15mg/kg) against triclabendazole (TCBZ)-resistant and TCBZ-susceptible F. hepatica infections. Activity against the Sligo TCBZ-resistant isolate was tested at three time points post-infection (p.i.): 3 days, 4 weeks and 12 weeks (Groups 1-3, respectively). A parallel trial was carried out using TCBZ (10mg/kg) (Groups 5-7): this provided a direct comparison between the efficacies of the two drugs. Group 4 served as an untreated Sligo control. Groups 8 and 9 were setup to test the efficacy of TCBZ and compound alpha against 12-week-old and 4-week-old TCBZ-susceptible, Cullompton infections, respectively. Group 10 served as an untreated Cullompton control. Sheep were sacrificed at 16 weeks p.i. and efficacies were determined. All remaining flukes were collected and measured, before being processed for whole-mount staining to assess the condition of their reproductive structures (testis, vitellaria, ovary and uterus). A second study was carried out to test the activity of compound alpha (15mg/kg) against mature 12-week-old TCBZ susceptible F. hepatica infections in sheep. Eighteen sheep were divided into two groups, A and B. Group A was treated and Group B served as an untreated control group. Efficacy was determined by reduction in faecal egg counts. The results showed that, whilst compound alpha was very active against adult TCBZ-susceptible flukes, producing a 100% reduction in faecal egg counts, it only caused a 62.5% reduction in fluke burden against juvenile flukes. Moreover, compound alpha was not effective against any stage of infection with TCBZ-resistant F. hepatica in sheep. Data from the trial also revealed biological differences between the two isolates. Thus, Sligo flukes were smaller in size and produced fewer eggs than the Cullompton flukes and their cysts were less infective to sheep. However, they reached the bile ducts more quickly and their eggs appeared in the faeces >2 weeks earlier. PMID- 19282109 TI - The schizotaxic self: phenotyping the silent predisposition to schizophrenia spectrum disorders. PMID- 19282110 TI - GNAI2 and regulators of G protein signaling as a potential Noonan syndrome mechanism. AB - Noonan syndrome (NS OMIM 163950) is a relatively common autosomal dominant developmental disorder characterized by short stature, specific facial features, and congenital cardiac anomalies. Approximately 50-66% of cases have defined mutations in the K-ras/Raf/MEK/ERK pathway that lead to constitutive signaling, but a significant number remain unexplained. We hypothesize that enhanced signaling through Galpha(i2) (from the GNAI2 gene) may also produce a NS-like phenotype. This is based on a recently described mouse model in which RGS mediated inhibition of Galpha(i2) is prevented by a knock-in mutation (G184S) that blocks RGS binding [Huang et al., Mol. Cell. Biol. 2006;26:6870-9]. The mice have short body length, cardiac hypertrophy, a triangular face with wide-set eyes and ears, and hematologic alterations. There is a slight increase in ERK activation and a pronounced enhancement of PI3K/Akt phosphorylation in MEFs from these mice suggesting that abnormal increases in Galpha(i2) signaling could represent a novel upstream mechanism for NS. This suggests a novel set of candidate genes for NS (GNAI2 and RGS proteins) and if validated could have important implications for therapy as well. PMID- 19282111 TI - Haplotype-tagging single nucleotide polymorphisms in the GSTP1 gene promoter and susceptibility to lung cancer. AB - BACKGROUND: Glutathione S-transferase (GST) P1 is a major phase II xenobiotic metabolizing enzyme in the human lung. Our laboratory had previously identified nine single nucleotide polymorphisms (SNPs) in the GSTP1 gene promoter, which were then grouped into three main haplotypes (Hap1, Hap2, and Hap3) based on statistical inference. Hap3 was found to display a high expression phenotype. The main objective of the current study was to test the association between GSTP1 promoter haplotypes with the risk of lung cancer after determining the promoter haplotypes experimentally through cloning and sequencing. METHODS: We conducted a case-control analysis of 150 subjects with lung cancer and 329 controls with no personal history of the disease. The three statistically inferred GSTP1 promoter haplotypes were confirmed experimentally through cloning and sequencing. Haplotype-tagging SNPs were selected and GSTP1 haplotypes were tested for genetic association to lung cancer using unconditional logistic regression after adjusting for confounders. Statistical interaction between GSTP1 promoter haplotypes with either cigarette smoking or dietary fruit and vegetable intake were tested using the likelihood ratio test. RESULTS: We did not find protective effects of Hap3 against lung cancer, despite an adequately powered design for this main effect. Homozygous variants of tagSNPs -1738 T>A and -354 G>T, which tag Hap2, showed an increased (but statistically non-significant) risk of lung cancer among all subjects as well as among individuals with low fruit and vegetable intake, compared to homozygous wildtypes for these SNPs. We did not find significant interactions between Hap2 and dietary intake of fruits and vegetables. CONCLUSIONS: Our results do not support significant main and modifying effects for GSTP1 promoter haplotypes on susceptibility to lung cancer in this population, but reinforce the protective effects of dietary intake of fruits and vegetables. PMID- 19282112 TI - [Fellowship in Canada: instructions]. AB - In North America, postdoctoral fellowships are proposed to physicians and surgeons after their residency to obtain an expertise in a specific domain of their speciality. In obstetrics and gynecology, three fellowship programs are accredited by the Royal College of Physicians and Surgeons of Canada: maternal fetal medicine, gynaecological oncology and reproductive endocrinology and infertility. A two-year fellowship in Canada provides a great professional and personal experience. We present here the organization of these programs and the conditions to be admitted in a fellowship program in Canada. PMID- 19282113 TI - In utero transmission of porcine torque teno viruses. AB - Sera and selected tissue homogenates collected from gnotobiotic swine never exposed to the environment or other swine tissues were tested for the presence of porcine torque teno virus (TTV) DNAs by nested and non-nested polymerase chain reactions (PCR) using primers specific for the untranslated region of porcine genogroups (g) 1 and 2. Twenty-three of 105 (21.9%) gnotobiotic piglets were g1- and/or g2-TTV DNA positive. Twenty-three of 27 (85.2%) sow sera, collected at the time of Caesarian derivation of the litters contained either or both TTV genogroup DNAs. These data demonstrate that porcine TTV may be transmitted to piglets by the in utero route and that the incidence of fetal infection is high. PMID- 19282114 TI - Hypomethylation trends in the intergenic region of the imprinted IGF2 and H19 genes in cloned cattle. AB - Bos taurus is a good model for embryo biotechnologies such as nuclear transfer. However, animals produced from these technologies often suffer from large calf syndrome, suggesting fetal growth dysregulation. The imprinted fetal mitogen IGF2 is clustered with H19 and the two genes are co-regulated in humans and mice. Although the allelic expression pattern of IGF2/H19 has been elucidated in agricultural species such as sheep and cattle, the underlying mechanism of their imprinting regulation has not been characterized. Using bisulfite sequencing the methylation status of 44 CpG sites in a CpG rich intergenic region of IGF2/H19 in the liver, brain, lung, kidney and placenta of control calves (produced by conventional breeding). One fragment containing 16 CpG sites was differentially methylated region (DMR), and thus may be important in regulating IGF2/H19 allelic expression. The DMR in tissues from cloned term calves that either died immediately after birth or were sacrificed due to complications shortly thereafter were examined. There were significant variations in the methylation of this DMR in some of the cloned animals compared to the controls. Most of the observed variations tended toward hypomethylation. The hypomethylation of this DMR in the liver and placenta of clones correlates with the previous observation of abnormal, biallelic expression of the H19 allele in those clones [Zhang, S., Kubota, C., Yang, L., Zhang, Y., Page, R., O'Neill, M., Yang, X., Tian, X.C., 2004. Genomic imprinting of H19 in naturally reproduced and cloned cattle. Biol. Reprod.] but not with allelic expression of IGF2 (as determined in this study). These data suggest that this DMR is involved in H19 allelic expression, but that other mechanisms probably regulate the expression of IGF2/H19. Contrary to global hypermethylation observed in cloned embryos, putative imprinting control regions can display hypomethylation trends in specific organs of cloned calves. PMID- 19282115 TI - Efficacy of a commercially available post-thaw bovine semen sexing kit in both single-ovulating and hyperstimulated cows. AB - Currently, there are few inexpensive, reliable, effective methods for commercially separating X- and Y-chromosome bearing fresh and frozen bovine sperm. The objective of these experiments was to determine the efficacy of a commercially available post-thaw bovine semen sexing kit, HeiferPlus (HP) which claims to alter the sex ratio in favor of female calves following artificial insemination. Three trials included the insemination of hyperstimulated cows with Control or HP-treated semen, non-surgical embryo collection on Day 7, and a combined PCR/dot blot assay to determine embryo sex. Chi-square analysis showed that the Control group produced a greater proportion (p<0.0005) of female embryos than the HP group. There were no differences in the proportions of transferable compared with degenerate embryos or in number of ovulations, embryos, and unfertilized ova collected from Control compared with HP groups. When treatments were combined, one of the two bulls used in the hyperstimulation studies produced an overall greater proportion of females (p<0.05), suggesting a bull effect. Another trial involved the insemination of cows synchronized via OvSynch((R)) with fetal sexing via ultrasonography. Results of these studies indicated that HP semen sexing kit did not alter the sex ratio in favor of females in either hyperstimulated or single-ovulating cows; however, potential bull effects may be further evaluated to understand the capacity of HP with semen from specific bulls. Additionally, perhaps the sex of the surviving embryo can be manipulated by the maternal side, through ovarian, hormonal, oviductal, or uterine influences. PMID- 19282116 TI - Changing course in ageing research: The healthy ageing phenotype. AB - Ageing is often associated with the aged and the diseased, nevertheless ageing is a process that starts in-uterus and is characterised by a progressive functional loss but not necessarily by the presence of disease and poor quality of life. How to meander through life without crossing the confines of major chronic disease and cognitive and physical impairment remains one of the most relevant challenges for science and humankind. Delimiting that 'immaculate' trajectory - that we dub as the 'Healthy Ageing Phenotype' - and exploring solutions to help the population to stay or return to this trajectory should constitute the core focus of scientific research. Nevertheless, current efforts on ageing research are mainly focused on developing animal models to disentangle the human ageing process, and on age-related disorders often providing merely palliative solutions. Therefore, to identify alternative perspectives in ageing research, Unilever and the Medical Research Council (MRC) UK convened a Spark workshop entitled 'The Healthy Ageing Phenotype'. In this meeting, international specialists from complementary areas related to ageing research, gathered to find clear attributes and definitions of the 'Healthy Ageing Phenotype', to identify potential mechanisms and interventions to improve healthy life expectancy of the population; and to highlight areas within ageing research that should be prioritised in the future. General agreement was reached in recognising ageing research as a disaggregated field with little communication between basic, epidemiological and clinical areas of research and limited translation to society. A more holistic, multi-disciplinary approach emanating from a better understanding of healthy ageing trajectories and centred along human biological resilience, its maintenance and the reversibility from early deviations into pathological trajectories, is urgently required. Future research should concentrate on understanding the mechanisms that permit individuals to maintain optimal health when facing pathological hazards and on developing and assessing potential interventions that could aid to re-establish resilience when lost or guarantee its integrity if present. Furthermore it is fundamental that scientific findings are translated incessantly into clear messages delivered to governmental institutions, the industry and society in general. PMID- 19282117 TI - Pre-analytical stability of selected benzodiazepines on a polymeric oral fluid sampling device. AB - Oral fluid field tests are designed to provide preliminary results with a high grade of reliability in order to meet analytical and forensic standards. Some test systems additionally offer the possibility of an independent confirmatory analysis of a test sample. The pre-analytical stability of 11 frequently abused benzodiazepines on an oral fluid collecting device (Drager DCD 5000) has been investigated. The collection device was designed to complement a special mobile testing system (Drager DrugTest 5000) to be sent to a laboratory for further confirmatory analysis. Blank oral fluid pool was spiked with a mixture of eleven frequently abused benzodiazepines and given onto a collection device. To simulate possible sample shipping, the collection devices were stored in the dark up to 14 days at ambient temperature in a plastic tube. The collection device was simultaneously stored without further treatment after oral fluid collection ('native') and with addition of 950muL of methanol, respectively. At different storage intervals repeat determination was carried out for every sample using a modified version of our standard LC-MS/MS method for the detection of benzodiazepines in serum. Different recoveries of benzodiazepines due to degradation and/or adsorption to the collection device during the 14 days of 'native' storage were found. Major loss of analytes was found for benzodiazepines containing a nitro-group such as flunitrazepam and clonazepam. This could be prevented almost completely by methanolic storage of the collection device after sampling. Therefore, we recommend the centrifugation of the collection device and separation from the polymer unit prior to sample shipping. If this should not be possible, addition of methanol immediately after sample collection can be used to avoid degradation of benzodiazepines during shipment. PMID- 19282118 TI - Chondrosarcoma of the nasal septum: A case report. AB - Chondrosarcoma make up only 10-20% of malignant primary bone tumors, with 5-10% located in the head and neck (Downey TJ, Clark SK, Moore DW. Chondrosarcoma of the nasal septum. Otolaryngol Head Neck Surg 2001;125:98-100), and nasal septal chondrosarcoma is extremely rare. Surgical excision is the only curative treatment and radiation and chemotherapy have a limited role for palliation. We used a navigation system in endoscopic surgery without complications for a case of chondrosarcoma of the nasal septum by means of a midfacial degloving approach at primary operation and an external incision approach at salvage operation for local recurrence. To our knowledge, this is the first report of the use of such a system for this surgical approach along with a salvage operation. We discuss the clinical presentation, diagnosis, and treatment of this case as well as present a review of the literature. PMID- 19282119 TI - [Immune reconstitution syndrome in the course of antiretroviral treatment in Senegal]. PMID- 19282120 TI - MR microscopy of micron scale structures. AB - Magnetic resonance (MR) microscopy of up to 1-5-microm resolutions have been reported previously. The tested phantom structures, however, had widths one order of magnitude bigger than the reported resolutions, e.g., spherical beads or capillary tubes of tens-of-micron diameters or wall thicknesses have been imaged. In this study, we fabricated structures having a few micron widths on a silicon wafer and imaged them using our 1-microm-resolution MR microscopy at 14.1 T. Micron scale width structures were, for the first time, resolved by MR microscopy. PMID- 19282121 TI - Assessment of the internal craniocervical ligaments with a new magnetic resonance imaging sequence: three-dimensional turbo spin echo with variable flip-angle distribution (SPACE). AB - PURPOSE: Lesions close to the internal craniocervical ligaments are a common problem in patients with whiplash injuries. The aim of this study was to evaluate the morphology and visibility of these ligamentous structures with a new isotropic three-dimensional (3D) turbo-spin-echo (TSE) technique. MATERIALS AND METHODS: MR (MR) images of the cervical spine of 52 healthy subjects (27 women and 25 men; mean age=29 years; age range=18-40 years) were taken with a T2 weighted 3D TSE sequence with variable flip-angle distribution [SPACE (Sampling Perfection with Application optimized Contrasts using different flip-angle Evolution)] at 1.5 T (Magnetom Avanto, Siemens Erlangen, Germany). Two experienced musculoskeletal radiologists read the images independently on a 3D imaging and postprocessing workstation. The visibility and morphology of the alar ligaments were evaluated on a five-point scale, and inter-reader correlation was assessed with kappa statistics. RESULTS: Both alar ligaments were detected in all subjects. Twenty-eight (53.8%) of the alar ligaments could not be seen within one slice of the standard coronal imaging plane but could adequately be visualized in an oblique reconstruction adapted to the orientation of the ligaments on the axial slices. Inter-reader correlation for visibility on MR imaging (MRI) of the internal craniocervical ligaments was high (left+right side, kappa=0.95). Most (94%) alar ligaments presented symmetrically. In the axial plane, 60% were oriented neutral and 40% had a backward orientation. In the coronal plane, 67% were oriented caudocranially and 33% were oriented horizontally. The shape of the ligaments was parallel in half and was V-shaped in the other half. The alar ligaments had homogeneous low-signal intensity in 56% and heterogeneous low signal intensity in 44%. The apical ligament of the dens was seen (excellent-good moderate) in 61% (reader 1) and 52% (reader 2). The tectorial membranes and the transverse ligament of the atlas were shown (excellent-good) in all subjects. CONCLUSIONS: MRI with acquisition of an isotropic SPACE technique allows high resolution imaging of the craniocervical ligaments in all orientations. Reconstruction of the image data in the variable orientation of the alar ligaments allowed for excellent depiction within one slice such that partial volume artifacts that hamper image analysis can be eliminated. PMID- 19282122 TI - Noninvasive monitoring of brain temperature during mild hypothermia. AB - The main purpose of this study was to verify the feasibility of brain temperature mapping with high-spatial- and reduced-spectral-resolution magnetic resonance spectroscopic imaging (MRSI). A secondary goal was to determine the temperature coefficient of water chemical shift in the brain with and without internal spectral reference. The accuracy of the proposed MRSI method was verified using a water and vegetable oil phantom. Selective decrease of the brain temperature of pigs was induced by intranasal cooling. Temperature reductions between 2 degrees C and 4 degrees C were achieved within 20 min. The relative changes in temperature during the cooling process were monitored using MRSI. The reference temperature was measured with MR-compatible fiber-optic probes. Single-voxel (1)H MRS was used for measurement of absolute brain temperature at baseline and at the end of cooling. The temperature coefficient of the water chemical shift of brain tissue measured by MRSI without internal reference was -0.0192+/-0.0019 ppm/degrees C. The temperature coefficients of the water chemical shift relative to N-acetylaspartate, choline-containing compounds and creatine were -0.0096+/ 0.0009, -0.0083+/-0.0007 and -0.0091+/-0.0011 ppm/degrees C, respectively. The results of this study indicate that MRSI with high spatial and reduced spectral resolutions is a reliable tool for monitoring long-term temperature changes in the brain. PMID- 19282123 TI - Feasibility of using limited-population-based average R10 for pharmacokinetic modeling of osteosarcoma dynamic contrast-enhanced magnetic resonance imaging data. AB - Retrospective analyses of clinical dynamic contrast-enhanced (DCE) MRI studies may be limited by failure to measure the longitudinal relaxation rate constant (R(1)) initially, which is necessary for quantitative analysis. In addition, errors in R(1) estimation in each individual experiment can cause inconsistent results in derivations of pharmacokinetic parameters, K(trans) and v(e), by kinetic modeling of the DCE-MRI time course data. A total of 18 patients with lower extremity osteosarcomas underwent multislice DCE-MRI prior to surgery. For the individual R(1) measurement approach, the R(1) time course was obtained using the two-point R(1) determination method. For the average R(10) (precontrast R(1)) approach, the R(1) time course was derived using the DCE-MRI pulse sequence signal intensity equation and the average R(10) value of this population. The whole tumor and histogram median K(trans) (0.57+/-0.37 and 0.45+/-0.32 min(-1)) and v(e) (0.59+/-0.20 and 0.56+/-0.17) obtained with the individual R(1) measurement approach are not significantly different (paired t test) from those (K(trans): 0.61+/-0.46 and 0.44+/-0.33 min(-1); v(e): 0.61+/-0.19 and 0.55+/ 0.14) obtained with the average R(10) approach. The results suggest that it is feasible, as well as practical, to use a limited-population-based average R(10) for pharmacokinetic modeling of osteosarcoma DCE-MRI data. PMID- 19282124 TI - Application of a rapid and selective method for the simultaneous determination of protease inhibitors, lopinavir and ritonavir in human plasma by UPLC-ESI-MS/MS for bioequivalence study in Indian subjects. AB - A high throughput and rugged ultra performance liquid chromatography tandem mass spectrometry (UPLC-ESI-MS/MS) method is developed and validated for the selective determination of protease inhibitors -- lopinavir (LPV) and ritonavir (RTV) in human plasma. Plasma samples were prepared by solid phase extraction of the analytes and their deuterated analogs as internal standard (IS) using Waters Oasis HLB cartridges. The chromatographic separation was achieved in a run time of 1.2 min on Waters Acquity UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm) under isocratic conditions. The mobile phase consisted of 10 mM ammonium formate, pH 4.0 adjusted with formic acid and methanol (10:90, v/v). The protonated precursor --> product ion transitions for lopinavir, ritonavir, d(8)-lopinavir and d(6)-ritonavir were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) and positive ion mode. A linear dynamic range of 2.9-1452 ng/mL and 29.6-14379 ng/mL was established for ritonavir and lopinavir respectively using 0.1 mL human plasma. The mean relative recovery of lopinavir (96.6%), ritonavir (97.5%), d(8)-lopinavir (85.5%) and d(6) ritonavir (86.3%) from spiked plasma samples was consistent and reproducible. The method was successfully applied to a bioequivalence study of [200(lopinavir)+50(ritonavir)]mg tablet formulation in 36 healthy human subjects under fasting conditions. PMID- 19282125 TI - [Continuous wound infiltration or intra-articular infusion for postoperative analgesia]. PMID- 19282126 TI - [Cascade hemofiltration: principle, first experimental data]. AB - High-volume hemofiltration has been suggested as an adjuvant treatment of septic shock (renal support and immunomodulation of the host response via the removal of middle molecular weight molecules such as cytokines). Nevertheless, high-volume hemofiltration presents some important drawbacks, such as the depletion of low molecular weight molecules (nutriments, vitamins, trace elements and antibiotics) due to the high ultrafiltration rate, or the significant financial cost and nursing workload. We describe cascade hemofiltration, a new high-volume hemofiltration system, which has been developed to limit these drawbacks by using a special extracorporeal circuit. Results of the first experimental study using this prototype are also reported. They demonstrate the technical feasibility, security and safety of the cascade system although other experimental and clinical studies are needed to continue evaluating this system. PMID- 19282127 TI - [Severe pulmonary contusion after extracorporeal shock wave lithotripsy]. AB - Extracorporeal shock wave lithotripsy (ESWL) is a simple and effective treatment of urinary stones. Renowned less aggressive than surgery, it knew a wide success and constitutes therefore the treatment of first intention of the majority of the kidney stones. Nevertheless, traumatic renal and extrarenal complications notably in lung can arise after ESWL. We report the case of a 28-year-old patient who had a pulmonary contusion following a lithotripsy for a left kidney stone and whose evolution was favourable after two weeks in intensive care unit. Through this observation and the analysis of the rare reported cases in the literature, we insist on the different varieties of pulmonary complications of the ESWL, the hypothesis explaining the mechanisms of their arising as well as the precautions to take to avoid them. PMID- 19282128 TI - [A complication of epidural anaesthesia: the breaking of a catheter]. AB - We report the breaking of an epidural catheter, which occurred during the procedure of insertion. If such a breaking is a rare event, it is due in this case to an aberrant trajectory of the catheter. By studying the literature, we also analyzed the mechanisms which can lead to the breaking of an epidural catheter. We recommend a precise radiological examination to localise the trapped catheter among the nervous structures. PMID- 19282129 TI - [Managing arterial hypertension: not so simple!]. PMID- 19282130 TI - [General practice has to contribute to the continuous improvement of quality of care and integrate health promotion]. AB - Quality of care in general practice is mainly based on effectiveness of clinical and inter-personal care, and access to care. Assessment has to focus on structures and processes, more than on outcomes. The sets of clinical indicators used in some european countries, though good starting points are still incomplete to encompass the complexity of activity. Assessment of care quality by the public authorities must be acceptable to the doctors who are the object of this assessment; otherwise its conclusions will be rejected. Continuous quality improvement can maintain quality of care at a high level and maintain costs under control. Health has to be managed, and doctors have a collective responsibility towards their patients. The procedures of the quality cycle are applicable in general practice for the development of community-based projects of health promotion, within the network of care. The discipline can find in it a new lease of life and perhaps curb the loss of interest among young doctors. PMID- 19282131 TI - [Airport malaria]. AB - Airport malaria is a particular form of autochthonous malaria: it happens when the Plasmodium infected Anopheles genus mosquito travels from an endemic area to a malaria free airport. Since 1969, 30 cases of airport malaria have been reported in France, 2 during summer 2008. The severity of airport malaria is explained by the frequency of Plasmodium falciparum infecting non immune individuals and an often important diagnosis delay. It is a compulsory notification disease in France. The International Health Regulations (IHR) require states to check that airplanes coming from malaria or arboviral endemic area are systematically disinsected. Vector control measures have to be implemented within a distance of at least 400 meters around the perimeter of airports in malaria or arboviral endemic areas. In France, this measure applies to all airports of French overseas territories, except for the island of Saint Pierre and Miquelon. PMID- 19282132 TI - [Jugular foramen syndrome and cervical varicella zoster viral infection]. PMID- 19282133 TI - [Mid-ventricular ballooning heart syndrome during drug withdrawal]. PMID- 19282134 TI - [Biologic therapies in the treatment of psoriasis]. AB - Psoriasis impairs significantly the quality of life. Systemic treatments have inconstant efficiency and dose-dependant toxicity. Since 2004, four biologic therapies - infliximab, etanercept, efalizumab and adalimumab - are approved in Europe for the treatment of moderate to severe psoriasis, with failure, intolerance or contra-indication to at least 2 "classical" systemic treatments, including phototherapy, methotrexate and ciclosporin. The estimated rate of patients reaching PASI75 at month 3 are as follow: about 80% under infliximab (classical W0-W2-W6 regimen); about 33% and 50% under etanercept, respectively, 25 and 50mg, twice weekly; about 25% under efalizumab (1mg/kg, weekly); about 70% under adalimumab (40mg, every other week). The 12-month efficiency of biologic therapies in psoriasis is poorly documented. The long term (5-year or more) safety of biologic therapies is not documented. New biologic therapies are under investigation in the treatment of psoriasis: ustekinumab (CNTO-1275), ABT-874, certolizumab (CDP-870), golimumab (CNTO-148). Ustekinumab is approved in Europe since January 2009. PMID- 19282135 TI - [What is biotherapy? The monoclonal antibody case]. AB - Biotherapy is the use of molecules derived from living organisms, cells, or even tissues for therapy. It is based on the exploitation of recent new insights of various biological mechanisms and relies heavily on a sophisticated cellular and molecular expertise. One of the major tools of biotherapy is currently monoclonal antibodies and derived products, whose are the subject of the present issue. Monoclonal antibodies have become major therapeutic drugs for treating a number of diseases, thanks to a remarkable molecular engineering. The success of the first generation of monoclonal antibodies opens the way to new challenges such as antibody functional optimization, better control of unwanted side effects, or low cost production at an industrial scale. A new generation of antibodies is now emerging and one can already foresee the future: oligoclonal approaches based on the use of specific antibodies cocktails, selection of eligible patients, antibody production at low costs... To date, up to 22 monoclonal antibodies are on the market, with more than two hundred being evaluated in clinical trials. PMID- 19282136 TI - [Intraparenchymal hemorrhage disclosing a primary angiitis of the central nervous system]. PMID- 19282137 TI - [Inflammatory myopathies: diagnosis and classifications]. AB - There are five major types of idiopathic inflammatory myopathies: dermatomyositis (DM), polymyositis (PM), inclusion body myositis (IBM), autoimmune necrotizing myopathy (AINM) and inflammatory myopathies associated with connective tissue diseases (overlap myositis). DM, PM and AINM are characterized by subacute, proximal and symmetrical weakness and respond to corticoids and immunosuppressants. Cutaneous involvement is specific for DM. IBM manifests by late onset, selective muscle weakness with early distal involvement and is unresponsive to immunosuppressants. PM is the rarest of these conditions. Histological features characterize each entity: perivascular inflammation, microangiopathy with reduced capillary density, ischemia, and perifascicular atrophy for DM; endomysial inflammation with invasion of non-necrotic fibers and diffuse expression of major histocompatibility complex class I antigens for PM; rimmed vacuoles in IBM coexisting with immunological features similar to PM; and necrosis is the prominent feature of AINM, without inflammation but associated with microangiopathy. The risk of malignant disease is increased in DM and AINM. Myopathy associated with anti-synthetase antibodies is characterized by frequent interstitial lung disease, perifascicular atrophy and prominent perimysial pathology. Myopathy associated with anti-SRP antibody is a necrotizing myopathy with rapid progression and partial resistance to corticoids. Inflammatory myopathies associated with connective tissue disease (CTD) are heterogeneous, involving all four major types (PM, DM, AINM, and IBM) and including additional pathological features. This category of myopathies has not yet been adequately characterized, because classification is usually replaced by the term "overlap myositis". PMID- 19282138 TI - Revision of the acetabulum with a contemporary cementless component. AB - We evaluated the performance of a contemporary cementless acetabular component at a minimum of 5 years postoperatively. One hundred eighty-seven consecutive acetabular component revisions were performed using a hemispherical porous-coated component. Patients were followed prospectively with radiographs and Harris hip scores. Twenty patients died, leaving 158 patients (166 hips) available for follow-up at a mean of 91 months. No patients were lost. Eleven acetabular components (7%) required repeated revision, including 4 (2%) for aseptic loosening. Seven of the 145 unrevised acetabular components with radiographic follow-up (5%) were loose. The results of acetabular revision with this contemporary acetabular component were good but inferior to those of earlier generation implants. This difference is likely multifactorial. PMID- 19282139 TI - The long-term results of the original Exeter polished cemented femoral component: a follow-up report. AB - We present a long-term follow-up report of the results of the original Exeter polished cemented stems inserted between November 1970 and the end of 1975 at our institution by surgeons of widely differing experience using crude cementing techniques. The results of this series were reported in 1988 (Fowler et al. Orthopaedic Clinics of North America. 1988;19:477) and again in 1993 (Timperley et al. Journal of Bone and Joint Surgery. 1993;75-B:33). There have been no failures from aseptic femoral component loosening since the 1993 report. From the original series of 433 hips, there were, at the end of 2003, 26 living patients with 33 hips. Of the latter, there were 25 hips in 20 patients with their original femoral components still in situ. Twenty-eight (6.46%) hips from the original series have been lost to follow-up, although none since 1992. With the end point reoperation for aseptic stem loosening, the survivorship is 93.5% (95% confidence interval, 90.0%-97.0%). The reoperation rate for aseptic femoral component loosening is 3.23% into the 33rd year of follow-up. PMID- 19282140 TI - Late repair of abductor avulsion after the transgluteal approach for hip arthroplasty. AB - The abductor release sometimes does not heal after a transgluteal approach for hip arthroplasty. Factors influencing the success of subsequent repair are unclear. We used magnetic resonance imaging (MRI) to compare the condition of the gluteus medius with clinical outcome after late repair of abductor dehiscence in 12 total hip patients. Evaluation included a pain rating, gait evaluation, Trendelenburg test, strength grading, and Harris Hip Score. Most had both prerepair and postrepair MRI studies to assess the repair and to grade abductor muscle fatty degeneration. Two repairs without MRI were explored surgically. Although average pain, limp, and strength scores improved significantly, rerupture occurred in 4 subjects and fatty degeneration in the gluteus medius did not improve, even with intact repair. Nine patients were satisfied; 7 of these had an intact repair. Magnetic resonance imaging and operative observations suggest that chronic degeneration in the abductor mechanism is the major impediment to successful repair. PMID- 19282141 TI - The structure of acute posttraumatic stress symptoms: 'reexperiencing', 'active avoidance', 'dysphoria', and 'hyperarousal'. AB - Empirical data have challenged chronic posttraumatic stress disorder (PTSD) consisting of three dimensions. In the present study we aimed to determine the factor structure of acute posttraumatic symptoms in two recently traumatized samples. In sample 1, 203 civilian trauma survivors were administered the Davidson Trauma Scale (DTS) approximately 1 week posttrauma. In sample 2, 182 civilian treatment seeking trauma survivors completed the DTS at an average of 41.4 days posttrauma. Our confirmatory factor analyses indicated that a 4-factor intercorrelated model provided the best representation of the data in both samples. The four factors are best described as reexperiencing, active avoidance, dysphoria, and hyperarousal. For acute posttraumatic symptoms, the empirical data suggest to split the avoidance cluster into 'Active avoidance' and 'Dysphoria' confirming findings in studies on chronic PTSD. In future revisions of the DSM, the diagnostic criteria for PTSD may need to be adapted to fit the research findings. PMID- 19282142 TI - The use of virtual reality in acrophobia research and treatment. AB - Acrophobia, or fear of heights, is a widespread and debilitating anxiety disorder affecting perhaps 1 in 20 adults. Virtual reality (VR) technology has been used in the psychological treatment of acrophobia since 1995, and has come to dominate the treatment of numerous anxiety disorders. It is now known that virtual reality exposure therapy (VRET) regimens are highly effective for acrophobia treatment. This paper reviews current theoretical understanding of acrophobia as well as the evolution of its common treatments from the traditional exposure therapies to the most recent virtually guided ones. In particular, the review focuses on recent innovations in the use of VR technology and discusses the benefits it may offer for examining the underlying causes of the disorder, allowing for the systematic assessment of interrelated factors such as the visual, vestibular and postural control systems. PMID- 19282143 TI - Relapse-remission and remission-relapse switches in rheumatoid arthritis patients were modeled by random effects. AB - OBJECTIVE: To compare statistical models for the analysis of two-state disease processes. STUDY DESIGN AND SETTING: A two-armed randomized trial of patients with early rheumatoid arthritis (RA) treated by either combination therapy (sulfasazaline, methotrextate, prednisolone) or monotherapy (sulfasazaline). Disease activity (remission or relapse) was analyzed with the logistic regression model, the proportional hazards regression model, and the continuous-time Markov process model for panel data. The dependence among the switching times was studied by (1) including correlated normal random patient effects for the relapse remission and remission-relapse switching probabilities; (2) assuming the population to be a mixture of patients responsive and nonresponsive to therapy; (3) including separate parameters for the first and subsequent relapse-remission switch; and (4) combining (1) and (3). The four approaches were compared using parametric bootstrap checks. RESULTS: The logistic regression model, the proportional hazards regression model, and the continuous-time Markov process model for panel data yielded similar combination therapy effects. The inclusion of random patient effects (approaches 1 and 4) gave the best fit to the observed disease activity pattern. CONCLUSION: Models with correlated random effects can provide a satisfactory fit to two-state disease patterns. PMID- 19282144 TI - Cochrane reviews used more rigorous methods than non-Cochrane reviews: survey of systematic reviews in physiotherapy. AB - OBJECTIVE: To describe the quality and methods of systematic reviews of physiotherapy interventions, compare Cochrane and non-Cochrane reviews, and establish the interrater reliability of the Overview Quality Assessment Questionnaire (OQAQ) quality assessment tool. STUDY DESIGN AND SETTING: A survey of 200 published systematic reviews was done. Two independent raters assessed the search strategy, assessment of trial quality, outcomes, pooling, conclusions, and overall quality (OQAQ). The study was carried out in the University research center. RESULTS: In these reviews, the five most common databases searched were MEDLINE, EMBASE, Cochrane Library, CINAHL, and Cochrane Review Group Registers. The Cochrane allocation concealment system and Jadad Scale were most frequently used to assess trial quality. Cochrane reviews searched more databases and were more likely to assess trial quality, report dichotomous outcomes for individual trials, and conduct a meta-analysis than non-Cochrane reviews. Non-Cochrane reviews were more likely to conclude that there was a beneficial effect of treatment. Cochrane reviews were of higher quality than non-Cochrane reviews. There has been an increase in the quality of systematic reviews over time. The OQAQ has fair to good interrater reliability. CONCLUSION: The quality of systematic reviews in physiotherapy is improving, and the use of Cochrane Collaboration procedures appears to improve the methods and quality. PMID- 19282145 TI - The Lower Extremity Functional Scale could be an alternative to the Western Ontario and McMaster Universities Osteoarthritis Index physical function scale. AB - OBJECTIVES: To compare the test-retest reliability and validity of the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) physical function (PF) subscale and the Lower Extremity Functional Scale (LEFS) in community dwelling adults with hip osteoarthritis (OA). STUDY DESIGN AND SETTING: One hundred adults with symptomatic hip OA participated in the study. Test-retest reliability was assessed by administering the WOMAC and LEFS at participants' initial visits and after 1 week. Discriminant validity from pain measures was assessed by examining the WOMAC-PF and the LEFS correlations with the PF and bodily pain subscales of the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36). Convergent validity was assessed by comparing correlations between the WOMAC-PF and LEFS with the gait speed test, step test, and timed stair tests. Intraclass correlation coefficients (ICCs), standard error of measurement, and Pearson correlation coefficients were calculated. RESULT: ICC estimates of 0.90 and 0.92 were obtained for the WOMAC-PF and LEFS, respectively. The minimal detectable change scores were 9.1 WOMAC-PF and 9.9 LEFS points. Discriminant validity was evident for the LEFS but not for the WOMAC-PF. The WOMAC-PF and LEFS showed similar levels of convergent validity; however, WOMAC-PF and gait speed showed a relatively low correlation. CONCLUSION: The LEFS has good measurement properties--namely test-retest reliability and cross-sectional construct validity--and it could be an alternative to WOMAC-PF. PMID- 19282146 TI - A new endpoint definition improved clinical relevance and statistical power in a vaccine trial. AB - OBJECTIVE: Endpoints used for the evaluation of immunogenicity in vaccine trials are often the proportion of individuals with immune response or geometric means of antibody concentrations for each serotype. When a vaccine includes several types of the same species, we illustrate how an endpoint combining all responses may improve clinical relevance and statistical power. STUDY DESIGN AND SETTINGS: The motivating example was the ANRS 114 Pneumovac trial where the effect of two vaccine strategies against Streptococcus pneumoniae was assessed in adults infected by the Human Immunodeficiency Virus. The power associated with several endpoints was calculated in the example and in simulations. A new endpoint based on four ordered levels is formulated and analyzed by using a proportional odds model. RESULTS AND CONCLUSION: The analysis of this new endpoint led to an odds ratio allowing detection of improvement and detriment. In the simulation study, this endpoint was associated with the largest statistical power by increasing the amount of information used as compared with usual endpoints. We recommend this new endpoint formulation in the formal development of a new vaccination regimen, whenever applicable. PMID- 19282147 TI - What constitutes progress in assessing patient outcomes? AB - Patient self-reported disability outcomes, measured using instruments such as the Health Assessment Questionnaire Disability Index (HAQ-DI) form a foundation for clinical studies of several conditions, notably arthritis. These instruments are "static" because the instrument is the same in all applications. Although generally performing well, they may fail at extremes of disability, the so-called floor and ceiling effects. Another limitation is the degree of measurement error, the signal-to-noise ratio. These two issues drive down the statistical power of clinical trials and increase their expense. PMID- 19282148 TI - Empirical evaluation showed that the Copas selection model provided a useful summary in 80% of meta-analyses. AB - OBJECTIVE: Although using meta-analysis to combine evidence from a number of studies should reduce both bias and uncertainty, it is sometimes not the case, because published studies represent a biased selection of the evidence. Copas proposed a selection model to assess the sensitivity of meta-analysis conclusions to possible selection bias. However, this relatively complex model awaits both reliable software and an empirical evaluation. This article reports work addressing both these issues. STUDY DESIGN AND SETTING: We took 157 meta-analyses with binary outcomes, analyzed each one using the Copas selection model, and evaluated each analysis using a prespecified protocol. The evaluation aimed to assess the usefulness of the Copas selection model to a typical Cochrane reviewer. RESULTS: In approximately 80% of meta-analyses, the overall interpretation of the Copas selection model was clear, with better results among the 22 with evidence of selection bias. However, as with the "Trim and Fill" method, allowing for selection bias can result in smaller standard errors for the treatment estimate. CONCLUSION: When a reliable test for selection bias is significant, we recommend systematic reviewers to try the Copas selection model, although the results should be interpreted cautiously. PMID- 19282149 TI - Integration of patient and provider assessments of mobility and self-care resulted in unidimensional item-response theory scales. AB - OBJECTIVE: The objective of this study was to develop a questionnaire that could integrate patient and provider items on mobility and self-care into unidimensional scales. The instrument should be suitable for various measurement models (patient and provider data [PAT-PRO], only patient data [PAT], only provider data [PRO]). STUDY DESIGN AND SETTING: The existing instruments, MOSES Patient and MOSES-Provider, were integrated into the MOSES-Combi and completed by a total of 1,019 neurology, cardiac, or musculoskeletal patients and/or their physicians (MOSES = acronym for "mobilty and self-care"). RESULTS: After selection of 18 items, all 12 scales of the MOSES-Combi (87 items) were largely unidimensional, met the standards for a 1-parameter item-response theory (IRT) model, were sufficiently reliable, and showed no differential item functioning (DIF) for age or gender. The person parameters set in the PAT-PRO measurement model show at least moderate, but usually substantial, agreement with those set in the PRO and PAT measurement models. CONCLUSION: The advantages of the MOSES Combi are that it can be used for various measurement models and is suitable for studying agreement between patient and provider assessments because of its psychometric properties (same scaling for patient and provider items). Integration of various data sources in an IRT scale can be extended to other assessments. PMID- 19282150 TI - Intraoral molluscum contagiosum imitating a squamous-cell carcinoma in an immunocompetent person--case report and review of the literature. AB - A case of intraoral molluscum contagiosum involving the right distal alveolar bridge of an immunocompetent person is reported. A review of the literature, with only five previously published cases, revealed that involvement of the oral soft tissues is rare. PMID- 19282151 TI - Modified trans-oral approach for mandibular condylectomy. AB - Different approaches to the mandibular condyle have been described. In this paper, a modified trans-oral technique to access the mandibular condyle is described and illustrated. This technique was used in a small group of patients; the clinical outcomes are promising. The technique can be used in various temporomandibular joint (TMJ) operations, such as condylar resection, high condylectomy or tumor removal. It provides adequate intra-oral surgical access to the mandibular condyle and avoids complications from extra-oral approaches to the TMJ. PMID- 19282152 TI - MDR1 gene in tardive dyskinesia scale scores: comparison of strategies for quantitative trait haplotype analysis. PMID- 19282153 TI - Ultrastructural and functional alterations of mitochondria in perilesional vitiligo skin. AB - BACKGROUND: Vitiligo is a chronic acquired hypomelanotic disorder affecting 0.5 2% of the world's population. The two major pathogenetic hypotheses are focused on immune-mediated or toxic-mediated cell damage primarily directed at melanocytes. Recent experimental data underline the complex interactions that exist between melanocytes and other cells found in the skin. OBJECTIVE: Among these cells, keratinocytes are able to influence both the survival and the functional activity of melanocytes. In order to gain insights into the involvement of different types of epidermic cells in the pathogenesis of vitiligo, we have performed an ultrastructural study on lesional, perilesional and normal skin from 12 patients. All these patients suffered from non-segmental vitiligo, with a similar clinical history in terms of lesion extension and duration of the disease. METHODS: We have therefore grown cultures of keratinocytes from lesional, perilesional and healthy skin, evaluating the presence of oxidative damage and apoptotic markers in the cells. RESULTS: Taken together, our results indicate that keratinocytes from perilesional skin show features of damaged cells. CONCLUSION: Our data, besides considering the achromic patch as the terminal event of a chain of biological processes that take place in the perilesional skin, highlight keratinocytes as having an important role in the development of vitiligo. PMID- 19282154 TI - Interactions between the glycine transporter 1(GlyT1) inhibitor SSR504734 and psychoactive drugs in mouse motor behaviour. AB - The specific glycine transporter 1 (GlyT1) inhibitor, SSR504734, is highly effective in enhancing N-methyl-D-aspartate receptor (NMDAR) function by elevating the availability of the NMDAR co-agonist, glycine, in the vicinity of NMDAR-containing glutamatergic synapses. According to the glutamatergic hypofunction hypothesis of schizophrenia, SSR504734 may therefore possess antipsychotic potential. Here, we evaluated the effects of SSR504734 in response to three psychomimetic drugs: phencyclidine, amphetamine, and apomorphine in male C57BL/6 mice. SSR504734 attenuated phencyclidine-induced (5 mg/kg, i.p.) hyperlocomotion, but potentiated the motor stimulant and motor depressant effects of amphetamine (2.5 mg/kg, i.p.) and apomorphine (0.75 mg/kg, s.c.), respectively. Hence, SSR504734 not only confers resistance to NMDAR blockade, but also enhances the locomotor response to dopaminergic stimulation. The latter finding adds to reports that SSR504734 may modulate dopamine-mediated behaviour by interference with normal glutamate-dopamine interaction. The specificity of this action of SSR504734 will be highly relevant to its potential application as an antipsychotic agent. PMID- 19282155 TI - Probing DNA hybridization efficiency and single base mismatch by X-ray photoelectron spectroscopy. AB - We demonstrated the use of X-ray photoelectron spectroscopy (XPS) to study DNA hybridization. Target DNA labeled with hexachloro-fluorescein (HEX) was hybridized to DNA arrays with four different probes. Each probe dot of the hybridized arrays was detected with XPS. The XPS Cl2p peak areas were found to decrease with an increase in mismatched bases in DNA probes. The Cl2p core-level peak area ratio of a probe perfectly matched to one, two and three base mismatched probes accorded well with the results of conventional fluorescent imaging, which shows that XPS is a potential tool for analyzing DNA arrays. The DNA arrays' hybridization efficiency was assessed by the molar ratio of chlorine to phosphorus in a DNA strand, which was determined from the relevant XPS Cl2p and P2p core-level peak areas after hybridization. This could provide a new method to detect DNA hybridization efficiency. PMID- 19282156 TI - WITHDRAWN: Cas radiologique du mois Radiological quiz of the month. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19282157 TI - Size does matter: can we reduce the radiotherapy field size for selected cases of anal canal cancer undergoing chemoradiation? AB - AIMS: Chemoradiation is the standard of care for the treatment of anal canal cancer, with surgery reserved for salvage. For tumours with uninvolved inguinal nodes, it is standard to irradiate the inguinal nodes prophylactically, resulting in large field sizes, which contribute to acute and late toxicity. The aim of this single-centre retrospective study was to determine if, in selected cases, prophylactic inguinal nodal irradiation could be avoided. MATERIALS AND METHODS: Between August 1998 and August 2004, 30 patients with biopsy-proven squamous cell anal canal cancer were treated with chemoradiation using one phase of treatment throughout. A three-field beam arrangement was used without attempting to treat the draining inguinal lymph nodes prophylactically. The radiotherapy dose prescribed was 50Gy in 25 daily fractions over 5 weeks. Concomitant chemotherapy was delivered with the radiation using mitomycin-C 7-12mg/m(2) on day 1 and protracted venous infusional 5-fluorouracil 200mg/m(2)/day throughout radiotherapy. RESULTS: All patients had clinically and radiologically uninvolved inguinal and pelvic nodes and all had primary lesions that were T3 or less. The median age at diagnosis was 65 years (range 41-84). The median follow-up was 41 months (range 24-113). The mean posterior field size was 14x15cm and the mean lateral field size was 12x15cm. All patients achieved a complete response. Ninety four per cent of patients (28/30) were alive and disease free. The two patients who died did so of unrelated causes and were disease free at death. Four patients relapsed and all were salvaged with surgery; two for local disease requiring abdominoperineal resection, one with an inguinal nodal relapse requiring inguinofemoral block dissection and one for metastatic disease to the liver who underwent liver resection. CONCLUSIONS: This single-centre retrospective study supports the treatment for selected cases of anal canal cancer with smaller than standard radiation fields, avoiding prophylactic inguinal nodal irradiation. Hopefully this will translate into reduced acute and late toxicity. In future studies we would suggest that consideration is given as to whether omission of prophylactic inguinal nodal irradiation for early stage tumours should be explored. PMID- 19282158 TI - Ultrasound imaging to assess inter- and intra-fraction motion during bladder radiotherapy and its potential as a verification tool. AB - AIMS: Organ motion is the principle source of error in bladder cancer radiotherapy. The aim of this study was to evaluate ultrasound bladder volume measurement as a surrogate measure of organ motion during radiotherapy: (1) to assess inter- and intra-fraction bladder variation and (2) as a potential treatment verification tool. MATERIALS AND METHODS: Twenty patients receiving radical radiotherapy for bladder cancer underwent post-void ultrasound bladder volume measurement at the time of radiotherapy treatment planning (RTP), and immediately before (post-void) and after receiving daily fractions. RESULTS: Ultrasound bladder volume measurement was found to be a simple and acceptable method to estimate relative bladder volume changes. Six patients showed significant changes to post-void bladder volume over the treatment course (P<0.05). The mean inter-fraction post-void bladder volume of five patients exceeded their RTP ultrasound bladder volume by more than 50%. Intra-fraction bladder volume increased on 275/308 (89%) assessed fractions, with the mean intra fraction volume increases of seven patients exceeding their RTP ultrasound bladder volume by more than 50%. CONCLUSIONS: Both day-to-day bladder volume variation and bladder filling during treatment should be considered in RTP and delivery. Ultrasound may provide a practical daily verification tool by: supporting volume limitation as a method of treatment margin reduction; allowing detection of patients who may require interventions to promote bladder reproducibility; and identifying patients with prominent volume changes for the selective application of more advanced adaptive/image-guided radiotherapy techniques. PMID- 19282159 TI - Hyperhomocysteinaemia and poor vitamin B status in chronic obstructive pulmonary disease. AB - BACKGROUND AND AIMS: Patients with chronic obstructive pulmonary disease (COPD) are at increased atherothrombotic risk. Preliminary findings have suggested that COPD patients may have increased plasma total homocysteine (tHcy), a cardiovascular risk factor often caused by a poor B vitamin status, but plasma levels of such vitamins were not measured. The aim of this study was to investigate hyperhomocysteinaemia in COPD and to determine whether it may be secondary to poor plasma concentrations of B vitamins. METHODS AND RESULTS: We performed a case-control, cross-sectional study of 42 patients with COPD and 29 control subjects. Folate, vitamin B12, vitamin B6, tHcy, renal function, C reactive protein, blood gases and lipids were measured in patients and controls. COPD patients had higher plasma tHcy (median: 13.9mumol/l, interquantile range [IQR]: 12.1-18.5 versus 11.5, IQR: 10.1-14, p=0.002) and lower circulating folate (median: 2.5ng/ml, IQR: 1.2-3.3 versus 2.8, IQR: 2.1-4 of controls, p=0.03) than controls had. Compared to the control group, COPD was associated with higher tHcy concentrations also after adjusting for smoking, heart failure, renal function and C-reactive protein with logistic regression analysis (OR 1.36, 95% CI 1.06 1.72, p=0.01). In the COPD group, low levels of folate (beta=-0.27, p=0.02) and vitamin B12 (beta=-0.24, p=0.04), and hypertriglyceridaemia (beta=0.580, p<0.0001) were independent predictors of the presence of high tHcy concentrations in a multiple linear regression model (adjusted R(2)=0.522). CONCLUSION: COPD patients have a poor B vitamin status and, as a consequence, increased tHcy. These abnormalities may contribute to the COPD-related atherothrombotic risk. PMID- 19282160 TI - Possible implications for health of the different definitions of dietary fibre. AB - There is impressive evidence from epidemiological and experimental studies that dietary fibre derived form vegetables, fruit and wholegrain cereals protects against and may be useful in the treatment of a wide range of diseases. However, while there is some evidence of benefit of extracted and synthetic fibres in terms of lowering levels of cardiovascular risk factors, improving measures of glycaemic control and gastrointestinal function, epidemiological confirmation of clinical benefit and long term safety are lacking. An appropriate definition of dietary fibre is essential, given that claiming a food is high in dietary fibre is in effect making a health claim, without formally doing so. The new Codex definition acknowledges the difference between naturally occurring carbohydrate polymers which are neither digested nor absorbed in the human small intestine and synthetic or extracted polymers. However the latter two groups may also be defined as dietary fibre provided "beneficial physiological effect has been demonstrated by generally accepted scientific evidence". Given the need for a definition of dietary fibre which can be used for food labelling, setting nutrient reference values and decisions relating to health claims it is important to achieve agreement as to what constitutes a meaningful physiological effect and the level of evidence required to be certain of such effect. PMID- 19282161 TI - Back to the ancient diet: a matter of urgency for Southern Mediterranean countries. PMID- 19282162 TI - A quantitative analysis of splint therapy of displaced temporomandibular joint disc. AB - OBJECTIVES: The effects of the Michigan splint on the change in disc displacement (DD) as well as the position of the condyles were determined by metrical analysis using magnetic resonance imaging (MRI). METHODS: Twenty-five patients with clinical DD symptoms were treated by means of the Michigan splint, and an assessment of the effects of the splint was conducted or verified by MRI before and during the period of therapy, 5 months follow-up. The positions of the condyles and the disc were calculated from the MRI in the parasagittal plane. RESULTS: There were no changes in the positions of the disc and condyles in the physiological joints of the patients (n=23) prior to and during the time the splint was in place (p>0.05). The splint achieved a DD decrease (p<0.05), and pain was eliminated in 69.2% of the DD joints with reduction (n=13). As far as the DD joints without reduction (n=13) are concerned, pain was eliminated in 74.9% of the joints, that is, without any change in the positions of the disc and condyles (p>0.05). CONCLUSIONS: The evaluation of the Michigan splint therapy showed that it has no influence in the repositioning of the DD joints without reduction, but the DD joints with reduction have a limited positive effect. In both forms of these displacements, conditions for the elimination of the clinical symptoms are created. PMID- 19282164 TI - WITHDRAWN: Endogenous lipoid pneumonia. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19282163 TI - Generation and maintenance of memory CD4(+) T Cells. AB - In the course of an immune response to an infectious microbe, pathogen-specific naive CD4(+) T cells proliferate extensively and differentiate into effector cells. Most of these cells die rapidly, but a small fraction of effector cells persist as memory cells to confer enhanced protection against the same pathogen. Recent advances indicate that strong TCR stimulation during the primary response is essential for the generation of long-lived memory CD4(+) T cells. Memory cells appear to be derived equally from all subsets of effector cells, and memory cells can also acquire additional functional capabilities during the secondary response. Resting memory CD4(+) cells are dependent on signals from contact with IL-7 and IL-15, but not MHC class II, for their survival and intermittent homeostatic proliferation. PMID- 19282165 TI - Biofiltration treatment of odors from municipal solid waste treatment plants. AB - An in situ compost biofilter was established for the treatment of odors from biostabilization processing of municipal solid waste. The concentrations of total volatile organic compounds (VOCs) in odors and their components were measured. Biofilter media was characterized in terms of total carbon (TC), total nitrogen (TN), total phosphorus (TP), organic matter (OM), pH value and determination of bacterial colony structure. Gas chromatography-mass spectrometry (GC-MS) analysis showed that the main components of the produced gas were benzene, toluene, ethylbenzene and xylene (BTEX) along with other alkanes, alkenes, terpenes, and sulphur compounds. The compost biofilter had remarkable removal ability for alkylated benzenes (>80%), but poor removal for terpenes ( approximately 30%). Total VOC concentrations in odors during the biostabilization process period ranged from 0.7 to 87 ppmv, and the VOC removal efficiency of the biofilter varied from 20% to 95%. After about 140 days operation, TN, TC, TP and OM in compost were kept almost stable, but the dissolved N, NH(4)-N and NO(3)-N experienced an increase of 44.5%, 56.2% and 76.3%, respectively. Dissolved P decreased by 27.3%. The pH value experienced an increase in the early period and finally varied from 7.38 to 8.08. Results of bacterial colony in packing material indicated that bacteria and mold colony counts increased, but yeasts and actinomyces decreased along with biofilter operation, which were respectively, 3.7, 3.4, 0.04 and 0.07 times of their initial values. PMID- 19282166 TI - Process parameters within a 750,000 litre anaerobic digester during a year of disturbed fermenter performance. AB - A 750,000litre fermenter was studied throughout one entire year by investigating the concentrations of volatile fatty acids (acetic, butyric, i-butyric, propionic, valeric and i-valeric acids), pH, concentrations of total C, N, S and NH(4)(+)-N, amounts of chemical and biological oxygen demand, and abundance of acetogenic microorganisms. Additionally several process parameters such as temperature, retention time, dry weight and input of substrate and liquids, and the concentrations and amounts of CH(4), H(2), CO(2) and H(2)S within the biogas were monitored continuously. Various volatile fatty acids and the ratio of acetic to propionic acid were shown to allow a rough indication on the fermentation but were not sufficiently precise to describe the fermenter performance. Nutrient compounds and special fractions, such as easily extractable carbohydrates or the concentration of total fats were more strongly correlated to the gas production of the fermenter. Results of an MPN-method for the determination of acetogenic microorganisms point to an important role of these microorganisms during the phase of restoration of the fermenter performance. PMID- 19282167 TI - Approaches to assess biocover performance on landfills. AB - Methane emissions from active or closed landfills can be reduced by means of methane oxidation enhanced in properly designed landfill covers, known as "biocovers". Biocovers usually consist of a coarse gas distribution layer to balance gas fluxes placed beneath an appropriate substrate layer. The application of such covers implies use of measurement methods and evaluation approaches, both during the planning stage and throughout the operation of biocovers in order to demonstrate their efficiency. Principally, various techniques, commonly used to monitor landfill surface emissions, can be applied to control biocovers. However, particularly when using engineered materials such as compost substrates, biocovers often feature several altered, specific properties when compared to conventional covers, e.g., respect to gas permeability, physical parameters including water retention capacity and texture, and methane oxidation activity. Therefore, existing measuring methods should be carefully evaluated or even modified prior to application on biocovers. This paper discusses possible strategies to be applied in monitoring biocover functionality. On the basis of experiences derived from investigations and large-scale field trials with compost biocovers in Austria, an assessment approach has been developed. A conceptual draft for monitoring biocover performance and recommendations for practical application are presented. PMID- 19282168 TI - Chiral recognition of mandelic acid by L-phenylalanine-modified sensor using quartz crystal microbalance. AB - This study presents a new method for the highly selective recognition of chiral mandelic acid (MA) using L-phenylalanine (L-Phe) as the selector. The proposed method is based on quartz crystal microbalance (QCM) detection, integrated with a vapor diffused molecular assembly (VDMA) reaction technique. The construction of the L-Phe-modified QCM sensor involved a two-step assembly procedure. The chiral recognizability of L- and D-MA on the L-Phe-modified surface was then examined using the VDMA method and QCM. The chiral discrimination factor between L- and D MA detected by QCM, alpha(L-MA/D-MA), was found to be about 8. The VDMA selective sensing of L-MA on the L-Phe-modified surface was also confirmed by the contact angle measurements. The L-Phe-modified QCM sensor showed good stability and reusability. The present chiral recognition results suggest that L-phe is an excellent resolving agent for the resolution of chiral mandelic acid. PMID- 19282169 TI - Clinical evaluation of continuous daily dosing of sunitinib malate in patients with advanced gastrointestinal stromal tumour after imatinib failure. AB - AIMS: To assess the antitumour activity, safety, pharmacokinetics and pharmacodynamics of continuous daily sunitinib dosing in patients with imatinib resistant/intolerant gastrointestinal stromal tumour (GIST) and to assess morning dosing versus evening dosing. PATIENTS AND METHODS: In this open-label phase II study, patients were randomised to receive morning or evening dosing of sunitinib 37.5mg/day. The primary end-point was clinical benefit rate (CBR; percent complete responses+partial responses [PRs]+stable disease [SD] 24 weeks). Secondary end-points included progression-free survival (PFS), overall survival (OS), safety, pharmacokinetic parameters and plasma biomarker levels. RESULTS: Sixty of 61 planned patients received treatment (30 per dosing group); 26 completed the study. Overall, the CBR was 53% (95% exact CI, 40-66): eight patients (13%) achieved objective PRs; 24 (40%) achieved SD 24 weeks. Median PFS was 34 weeks (95% CI, 24-49); median OS was 107 weeks (95% CI, 72 - not yet calculable). Most adverse events (AEs) were of grade 1 or 2 in severity, and were manageable through dose modification or standard interventions. No new AEs were apparent compared with the approved intermittent dosing schedule. Antitumour activity and safety were generally similar with morning and evening dosing. Continuous daily sunitinib dosing achieved and sustained effective drug concentrations without additional accumulation across cycles. Decreases from baseline in plasma levels of soluble KIT after 20 and 24 weeks of dosing correlated with longer OS. CONCLUSION: For patients with imatinib resistant/intolerant GIST, continuous daily sunitinib dosing appears to be an active alternative dosing strategy with acceptable safety. PMID- 19282170 TI - Indications and timing for prostate biopsy, diagnosis of early stage prostate cancer and its definitive treatment: a clinical conundrum in the PSA era. AB - The use of PSA for prostate cancer screening has led to a large increase in the number of men undergoing transrectal ultrasonography (TRUS) - guided biopsy of the prostate to determine the presence of prostate cancer. Recently, the indications for prostate biopsy based on PSA level have been questioned and new prostate cancer risk calculators that consider other factors related to prostate cancer have been proposed. Also, there have been significant changes over the years regarding the protocols used to sample the prostate. Most protocols recommend more extensive sampling of the prostate with more laterally directed biopsies of the peripheral zone for both initial and subsequent biopsies. There is still much controversy over the appropriate number and location of biopsy cores, and timing to optimize the diagnosis of prostate cancer on initial and repeat biopsy. Finally, discovery of a new molecular marker independent of the PSA level will be very important in the diagnosis and prognosis of prostate cancer. PMID- 19282171 TI - Inhibitors of potassium channels KV1.3 and IK-1 as immunosuppressants. AB - New structural classes of K(V)1.3 and IK-1 ion channel blockers have been identified based on a virtual high throughput screening approach using a homology model of KcsA. These compounds display inhibitory effects on T-cell and/or keratinocyte proliferation and immunosuppressant activity within a DTH animal model. PMID- 19282172 TI - Synthesis of protein tyrosine phosphatase 1B inhibitors: model validation and docking studies. AB - The designed and synthesized 2-(4-methoxyphenyl) ethyl] acetamide derivatives (3a, 3b and 3c) were evaluated for their PTP1B inhibitory activity where they showed IC(50) values 69 microM, 87 microM and 71 microM, respectively. These results correlated well with the docking studies and in vivo screening of the compounds for their antidiabetic activity in SLM and STZ models. PMID- 19282173 TI - Novel small molecule activators of beta-catenin-mediated signaling pathway: structure-activity relationships of indirubins. AB - Based on the beta-catenin-drived Wnt activator of bromoindirubin-3'-oxime (BIO), indirubin analogs were evaluated for beta-catenin-mediated gene expression. A novel indirubin analog, indirubin-5-nitro-3'-oxime (INO), was considered a potential activator, and structure-activity studies were conducted with indirubins. These data suggest that INO might be a novel Wnt activator and has a potential of signaling regulator in beta-catenin-mediated signaling pathways. PMID- 19282174 TI - Design and optimization of selective serotonin re-uptake inhibitors with high synthetic accessibility. Part 1. AB - The reported selective serotonin Re-uptake Inhibitor Litoxetine was used as the starting point in the design of a range of potential SSRIs with high ease of synthetic accessibility. Preparation and subsequent optimization yielded a range of potent and highly selective SSRIs. PMID- 19282175 TI - Synthesis and evaluation of arylalkoxy- and biarylalkoxy-phenylamide and phenylimidazoles as potent and selective sphingosine-1-phosphate receptor subtype 1 agonists. AB - In pursuit of potent and selective sphingosine-1-phosphate receptor agonists, we have utilized previously reported phenylamide and phenylimidazole scaffolds to explore extensive side-chain modifications to generate new molecular entities. A number of designed molecules demonstrate good selectivity and excellent in vitro and in vivo potency in both mouse and rat models. Oral administration of the lead molecule 11c (PPI-4667) demonstrated potent and dose-responsive lymphopenia. PMID- 19282176 TI - QSAR analysis of pyrazolidine-3,5-diones derivatives as Dyrk1A inhibitors. AB - Individuals with Down syndrome (DS) suffer from mental retardation. Overexpression and the resulting increased specific activity of Dyrk1A kinase located on chromosome 21 cause a learning and memory deficit in Dyrk1A transgenic mice. To search for therapeutic agents with Dyrk1A inhibition activity, previously we obtained HCD160 as a new hit compound for Dyrk1A inhibition. In the present study, we synthesized 34 HCD160 derivatives to investigate the quantitative structure-activity relationship (QSAR). This analysis could provide important information for novel drug discovery for treatment of DS related learning and memory deficits. PMID- 19282177 TI - Syntheses of novel high affinity ligands for opioid receptors. AB - A series of novel high affinity opioid receptor ligands have been made whereby the phenolic-OH group of nalbuphine, naltrexone methiodide, 6-desoxonaltrexone, hydromorphone and naltrindole was replaced by a carboxamido group and the furan ring was opened to the corresponding 4-OH derivatives. These furan ring 'open' derivatives display very high affinity for mu and kappa receptors and much less affinity for delta. The observation that these target compounds have much higher receptor affinity than the corresponding ring 'closed' carboxamides significantly strengthens our underlying pharmacophore hypothesis concerning the bioactive conformation of the carboxamide group. PMID- 19282178 TI - Primary medulla oblongata germinoma--an unusual posterior fossa tumors in young adults. AB - We encountered 2 patients with germinoma arising from the medulla oblongata in whom preoperative radiological diagnosis was difficult. A 30-year-old woman presented due to aspiration pneumonia caused by bilateral lower cranial nerve palsies, and a 24-year-old man presented with headache caused by obstructive hydrocephalus. In both patients, there was a midline tumor that extended from the lower part of the fourth ventricle to the C1 lamina level. It was well-demarcated and homogeneously enhanced tumor with a slightly high density on plain CT scan. The preoperative diagnosis for both patients was ependymoma. The former patient had persistent lower cranial nerve palsies due to brain stem injury after tumor resection. Both patients achieved complete remission with adjuvant therapy. Fewer than 10 cases of germinoma affecting the medulla oblongata have been reported. Radiological findings resembling those of the pineal region germinoma were observed in the two patients reported here. We would like to stress the importance of remembering germinoma when making a preoperative differential diagnosis of fourth ventricular tumors in young adults. PMID- 19282179 TI - Asymmetric dimethylarginine and nitric oxide levels in migraine during the interictal period. AB - Nitric oxide (NO), which modulates endothelial function, is thought to be pivotal in the pathophysiology of migraines. The connection between migraine and cardiovascular diseases has also drawn attention to the endothelial dysfunctions and NO pathway abnormalities seen in patients with migraine. Our goal was to assess the levels of NO and the endogenous NO synthase inhibitor, asymmetric dimethylarginine (ADMA), in people with migraine during the interictal period. A total of 49 patients with migraine and 22 control subjects were enrolled in the study. Their plasma NO metabolites (nitrite [NO2-] and nitrate [NO3-]) and ADMA levels were measured using the enzyme-linked immunosorbent assay method, and were then compared with their cardiovascular risk factors, anthropometric measurements, and headache frequency and severity. The plasma ADMA, NO2- and NO3- levels of the patients with migraine during the interictal period did not differ from the control group, and no relationship was found between cardiovascular risk factors and migraine attack severity and frequency. We conclude that, in patients with migraine, there is no dysfunction of baseline NO and ADMA metabolism during the interictal period. PMID- 19282180 TI - Cauda equina syndrome following a lumbar puncture. AB - Lumbar puncture (LP), a common diagnostic procedure, is usually associated with low morbidity. We describe the case of a 29-year-old woman who underwent a non traumatic LP in the setting of normal coagulation. Cauda equina syndrome subsequently developed secondary to an extradural spinal haematoma. Avoidance, identification and management of this uncommon complication are discussed. Iatrogenic cauda equina syndrome following LP is rare, but can cause significant morbidity. Our patient's experience and our review of the literature highlight that: (i) normal coagulation and a non-traumatic LP do not exclude this diagnostic possibility; (ii) early recognition determines the management and prognosis, as 50% of patients remain paraplegic if the condition is identified more than 12 hours after symptom onset; and (iii) neurosurgical intervention can be avoided, despite bladder dysfunction, if there are early signs of recovery. PMID- 19282181 TI - PUPylation: something old, something new, something borrowed, something Glu. AB - Most eukaryotic proteins are degraded by the 26S proteasome as a consequence of their covalent modification with ubiquitin. Although the proteasome is found in some prokaryotes, ubiquitin is not, which indicates that substrates are targeted to prokaryotic proteasomes by a fundamentally different mechanism. A recent study has identified Pup (prokaryotic ubiquitin-like protein) as a mycobacterial protein that functions in a manner analogous to ubiquitin for proteasome dependent proteolysis in prokaryotes. PMID- 19282182 TI - H2A.Z and DNA methylation: irreconcilable differences. AB - DNA methylation state and the composition of the nucleosome core particle influence chromatin structure and, in turn, transcriptional competence. Although it is clear that chromatin remodeling and covalent histone modifications regulate DNA methylation in plants and animals, the role of histone variants in directing DNA methylation, and vice versa, has not been addressed. A new genome-wide study in Arabidopsis thaliana reveals a broadly antagonistic relationship between H2A.Z occupancy and DNA methylation. PMID- 19282183 TI - Sumoylation and human disease pathogenesis. AB - Covalent modification by SUMO polypeptides, or sumoylation, is an important regulator of the functional properties of many proteins. Among these are several proteins implicated in human diseases including cancer, Huntington's, Alzheimer's, and Parkinson's diseases, as well as spinocerebellar ataxia 1 and amyotrophic lateral sclerosis. Recent reports reveal two new examples of human disease-associated proteins that are SUMO modified: amyloid precursor protein and lamin A. These findings point to a function for sumoylation in modulating amyloid beta peptide levels, indicating a potential role in Alzheimer's disease, and for decreased lamin A sumoylation as a causative factor in familial dilated cardiomyopathy. PMID- 19282184 TI - 8-Bromo-9-alkyl adenine derivatives as tools for developing new adenosine A2A and A2B receptors ligands. AB - Importance of making available selective adenosine receptor antagonists is boosted by recent findings of adenosine involvement in many CNS dysfunctions. In the present work a series of 8-bromo-9-alkyl adenines are prepared and fully characterized in radioligand binding assays or functional cyclase experiments in respect to their interaction with all the four adenosine receptor subtypes. Results show that the presence of the bromine atom in 8-position of 9-substituted adenines promotes in general the interaction with the adenosine receptors, in particular at the A(2A) subtype. The present study also demonstrates that adenine derivatives could be a good starting point to obtain selective adenosine A(2B) receptor antagonists. PMID- 19282185 TI - Synthesis and evaluation of myxochelin analogues as antimetastatic agents. AB - Myxochelin A (1) is an inhibitor of tumor cell invasion produced by the bacterium belonging to the genus Nonomuraea. In order to obtain more potent inhibitors, a series of myxochelin analogues [2 and (S)-3-17] were synthesized through the coupling of lysine or diaminoalkane derivatives and appropriately protected hydroxybenzoate, followed by modification of functional groups and deprotection. These compounds were evaluated for their inhibitory activity against invasion of murine colon 26-L5 carcinoma cells. Among the synthetic analogues tested, compound (S)-6 which possesses carbamoyl group at C-1 was found to be the most potent antiinvasive agent and is considered to be a promising lead molecule for the antimetastasis. Compound (S)-6 was also shown to inhibit gelatinase activities of MMP-2 and MMP-9 and in vivo lung metastasis in mice. PMID- 19282186 TI - Antiproliferative and antimalarial anthraquinones of Scutia myrtina from the Madagascar forest. AB - Bioassay-guided fractionation of an ethanol extract of a Madagascar collection of the bark of Scutia myrtina led to the isolation of three new anthrone anthraquinones, scutianthraquinones A, B and C (1-3), one new bisanthrone anthraquinone, scutianthraquinone D (4), and the known anthraquinone, aloesaponarin I (5). The structures of all compounds were determined using a combination of 1D and 2D NMR experiments, including COSY, TOCSY, HSQC, HMBC, and ROESY sequences, and mass spectrometry. All the isolated compounds were tested against the A2780 human ovarian cancer cell line for antiproliferative activities, and against the chloroquine-resistant Plasmodium falciparum strains Dd2 and FCM29 for antiplasmodial activities. Compounds 1, 2 and 4 showed weak antiproliferative activities against the A2780 ovarian cancer cell line, while compounds 1-4 exhibited moderate antiplasmodial activities against P. falciparum Dd2 and compounds 1, 2, and 4 exhibited moderate antiplasmodial activities against P. falciparum FCM29. PMID- 19282187 TI - Binary and ternary inclusion complexes of finasteride in HPbetaCD and polymers: preparation and characterization. AB - The aim of this study was to determine whether inclusion complexes between 2 hydroxypropyl-beta-cyclodextrin (HPbetaCD) and finasteride (FIN) are formed, and to characterize these. Equimolar FIN/HPbetaCD solid systems in the presence or absence of 0.1% (w/v) of polyvinylpyrrolidone K30 (PVP K30) or 0.3% of chitosan were prepared by coevaporation and freeze-drying methods. The systems were characterized by phase solubility, NMR, DSC, and XRD analysis. The results suggest that true binary and ternary inclusion complexes were formed. PMID- 19282188 TI - Discovery and structure-activity relationships of (2 (arylthio)benzylideneamino)guanidines as a novel series of potent apoptosis inducers. AB - 1-(2-(2,5-Dimethoxyphenylthio)benzylidene)semicarbazide (2a) was discovered as a potent apoptosis inducer through our cell based HTS assay. SAR study led to the discovery of a more aqueous soluble analog (2-(2,5-dimethoxyphenylthio)-6 methoxybenzylideneamino)guanidine (5e) with EC(50) value of 60 nM in the caspase activation assay and GI(50) value of 62 nM in the growth inhibition assay in T47D cells. Compound 5e was found to be an inhibitor of tubulin polymerization and efficacious in a MX-1 breast tumor model. PMID- 19282189 TI - Discovery of potent and orally active 3-alkoxy-5-phenoxy-N-thiazolyl benzamides as novel allosteric glucokinase activators. AB - Identification and synthesis of novel 3-alkoxy-5-phenoxy-N-thiazolyl benzamides as glucokinase activators are described. Removal of an aniline structure of the prototype lead (2a) and incorporation of an alkoxy or phenoxy substituent led to the identification of 3-Isopropoxy-5-[4-(methylsulfonyl)phenoxy]-N-(4-methyl-1,3 thiazol-2-yl)benzamide (27e) as a novel, potent, and orally bioavailable GK activator. Rat oral glucose tolerance test indicated that 27e exhibited a glucose lowering effect after 10 mg/kg oral administration. PMID- 19282190 TI - Discovery of 2-(5-nitrothiazol-2-ylthio)benzo[d]thiazoles as novel c-Jun N terminal kinase inhibitors. AB - A new series of 2-thioether-benzothiazoles has been synthesized and evaluated for JNK inhibition. The SAR studies led to the discovery of potent, allosteric JNK inhibitors with selectivity against p38. PMID- 19282191 TI - Thiodisaccharides with galactofuranose or arabinofuranose as terminal units: synthesis and inhibitory activity of an exo beta-D-galactofuranosidase. AB - Thiodisaccharides having beta-D-Galf or alpha-L-Araf units as non-reducing end have been synthesized by the SnCl(4)- or MoO(2)Cl(2)-promoted thioglycosylation of per-O-benzoyl-D-galactofuranose (1), its 1-O-acetyl analogue 4, or per-O acetyl-alpha-L-arabinofuranose (16) with 6-thioglucose or 6-thiogalactose derivatives. After convenient removal of the protecting groups, the free thiodisaccharides having the basic structure beta-D-Galf(1-->6)-6-thio-alpha-D Glcp-OMe (5) or beta-D-Galf(1-->6)-6-thio-alpha-D-Galp-OMe (15) were obtained. The respective alpha-L-Araf analogues 18 and 20 were prepared similarly from 16. Alternatively, beta-D-Galf(1-->4)-4-thio-3-deoxy-alpha-L-Xylp-OiPr was synthesized by Michael addition to a sugar enone of 1-thio-beta-d-Galf derivative, generated in situ from the glycosyl isothiourea derivative of 1. The free S-linked disaccharides were evaluated as inhibitors of the beta galactofuranosidase from Penicillium fellutanum, being 15 and 20 the more active inhibitors against this enzyme. PMID- 19282192 TI - Aryl nucleoside H-phosphonates. Part 16: synthesis and anti-HIV-1 activity of di aryl nucleoside phosphotriesters. AB - Di-aryl nucleoside phosphotriesters have been explored as a new type of pronucleotides for the purpose of anti-HIV-1 therapy and efficient synthetic protocols, based on H-phosphonate chemistry, have been developed for the preparation of this class of compounds. It was found that anti-HIV-1 activity of the phosphotriesters bearing an antiviral nucleoside moiety (AZT, ddA) and also ddU was due, at least partially, to intracellular conversion into the corresponding nucleoside 5'-monophosphates, and their efficiency correlated well with the pK(a) values of the aryloxy groups present. PMID- 19282193 TI - Experimental determination of proton-induced cross-sections on natural zirconium. AB - We measured cross-sections for the formation of (86g,87m,87g)Y, (88,89g)Zr, and (90,92m)Nb radionuclides for proton-induced reactions on natural zirconium by using a conventional stacked-foil activation technique in the energy range between 1 and 40MeV at the MC-50 cyclotron of the Korea Institute of Radiological and Medical Science. We compared the measured data with the available literature data and the theoretical calculation by the model codes TALYS and ALICE-IPPE. We also deduced the integral yields for thick targets from the measured cross sections of the produced radionuclides. The optimum production possibility of the medically important (89g)Zr and (86)Y radionuclides were discussed elaborately. PMID- 19282194 TI - Development of a new design (125)I-brachytherapy seed for its application in the treatment of eye and prostate cancer. AB - Palladium coated silver beads of 0.5mm (slashed circle) were used to adsorb (125)I and encapsulated inside a titanium capsule by Nd:YAG laser, for use as a brachytherapy source. Experimental conditions were optimized for maximum adsorption of (125)I and uniformity of radioactivity was ascertained. Leachability of (125)I was found to be<0.01%. The dose rate constant of the new source was estimated to be 1.045cGyh(-1)U(-1). This newly developed source could be an alternative to other (125)I sources. PMID- 19282195 TI - Development of position sensitive radiation detectors using gas electron multipliers. AB - Gas electron multipliers (GEM) were introduced to develop a radiation detector which is applicable to medical imaging or luggage inspection systems at the airport or harbor. Two GEM foils were used in the amplifier, and an Ar/CO(2) mixed gas was inserted into the chamber at a mixing ratio of Ar:CO(2)=80:20. A two-dimensional X-ray image was taken with a 64-channel GEM detector from an Fe 55 radiation source. We also constructed a 256-channel GEM detector in which 4 charge sensitive preamplifiers were used in a daisy chain. With linear array type readout electrodes, we were able to realize a position sensitive radiation detector. PMID- 19282196 TI - Syringe shape and positioning relative to efficiency volume inside dose calibrators and its role in nuclear medicine quality assurance programs. AB - A careful analysis of geometry and source positioning influence in the activity measurement outcome of a nuclear medicine dose calibrator is presented for (99m)Tc. The implementation of a quasi-point source apparent activity curve measurement is proposed for an accurate correction of the activity inside several syringes, and compared with a theoretical geometric efficiency model. Additionally, new geometrical parameters are proposed to test and verify the correct positioning of the syringes as part of acceptance testing and quality control procedures. PMID- 19282197 TI - Preparation and bioevaluation of a 99mTc-labeled chlorambucil analog as a tumor targeting agent. AB - Chlorambucil belongs to a group of nitrogen mustards which are used for the treatment of variety of cancers. Hence, a chlorambucil derivative has been radiolabeled with [(99m)Tc(CO)(3)(H(2)O)(3)](+) core and its efficacy as a tumor targeting agent has been evaluated. Radiochemical yield of the complex was >98% as observed by HPLC. The in vitro studies in MCF-7 breast cancer cells showed about 30% inhibition of the radiolabeled complex in presence of the cold chlorambucil derivative. Biodistribution studies in Swiss mice bearing fibrosarcoma tumor showed an uptake of 3.2+/-0.3%ID/g at 3h.p.i. PMID- 19282198 TI - Radioactivity and fluoride contamination derived from a phosphate fertilizer plant in Egypt. AB - The environmental pollution caused by the wastewater from a phosphate fertilizer plant in Egypt was investigated. The concentrations of radionuclides and fluoride in phosphate fertilizer (raw materials, end products and by-products) and other types of fertilizer samples were measured. The concentrations of these elements were also measured in environmental samples (water, sediment and plant) collected from the proximity of outlet of wastewater discharge pipes of the phosphate fertilizer company. The fluoride concentration was ranged from 0.03 to 0.25mg/g, 0.002 to 0.006mg/g, 0.42 to 1.88mg/g and 0.44 to 7.3mg/l for phosphate fertilizer, other types of fertilizer, sediment and water samples, respectively. The activity concentrations of (226)Ra were ranged from 244 to 1312Bq/kg, 0.6 to 12.1Bq/kg, 15.4 to 33.8Bq/kg, 0.06 to 1.3Bq/l and 8.9 to 17.3Bq/kg for phosphate fertilizer, other types of fertilizer, sediment, water and plant samples, respectively. The (232)Th activity concentrations were ranged from 0.7 to 24Bq/kg, 0.7 to 14.5Bq/kg, 10.4 to 19.3Bq/kg, 0.02 to 0.16Bq/l and 2.0 to 29.8Bq/kg for these samples, respectively. Also, the (40)K activity concentrations were ranged from 2.1 to 1.4Bq/kg, 2.1 to 5313Bq/kg, 128 to 281Bq/kg, 0.14 to 0.6Bq/l and 686 to 977Bq/kg for these samples, respectively. Low content of (137)Cs was determined in only two phosphate fertilizer samples (F2 and F3; mean 1.3Bq/kg) and in most of sediment samples (with range 1.0 2.4Bq/kg). The radium equivalent, as a radiation hazard index, was ranged from 284 to 1316, 9.6 to 432 and 47 to 70Bq/kg for phosphate fertilizer, other types of fertilizer and sediment samples, respectively. The results indicated that the wastewater polluted with fluoride produced from the phosphate fertilizer company may be affecting the environment. The radioactivity content measurements indicated that the environment may be slightly affected with low concentrations of (226)Ra and (232)Th isotopes due to the discharged wastewater from the phosphate fertilizer industry. On the other hand, the results of comparison studies for radioactivity concentrations are also presented and discussed. PMID- 19282199 TI - Effect of trifluoroacetate, a persistent degradation product of fluorinated hydrocarbons, on Phaseolus vulgaris and Zea mays. AB - The aim of this study was to quantify the effect of the pollutant, trifluoroacetate (TFA), on growth and photosynthesis of Phaseolus vulgaris (C(3)) and Zea mays (C(4)) in order to elucidate the physiological and biochemical basis of its inhibitory action. In whole plant studies, photosynthetic gas exchange, fast phase fluorescence kinetics and Rubisco activity were measured in parallel over a 14-day period in plants cultivated in a water culture system with NaTFA added at concentrations ranging from 0.625 to 160mgl(-1). Although initial stimulation of some photosynthetic parameters was observed at low TFA concentrations early on in the experiment, marked inhibition occurred at higher concentrations. In general Z. mays was affected more severely than P. vulgaris showing a large TFA-induced decrease in both apparent carboxylation efficiency (ACE) and in vitro Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase; EC 4.1.1.39) activity. Analysis of photosynthetic gas exchange revealed that besides constraints on mesophyll processes such as Rubisco activity, stomatal limitation also increased with increasing TFA concentration, especially in P. vulgaris. In depth analysis of the fast phase fluorescence transients pointed at TFA-induced uncoupling of the oxygen evolving complex (OEC) and inhibition of electron transport beyond Q(a) including possible constraints on the reduction of end electron acceptors of photosystem I. PMID- 19282200 TI - Vitamin D and melanoma. AB - PURPOSE: Ultraviolet light from sunlight and other sources is the major environmental risk factor for melanoma of the skin. Humans also derive most of their vitamin D from exposure to sunlight. This article reviews current evidence that vitamin D might play a preventive role in the development of melanoma or affect tumor aggressiveness or melanoma patient outcomes. METHODS: Literature review. RESULTS: The vitamin D receptor has been identified in normal melanocytes as well as melanoma cell lines and primary tissue. A few studies have demonstrated relationships of functional polymorphisms in the vitamin D receptor with melanoma risk or tumor aggressiveness. Identifying an independent influence of vitamin D on melanoma risk is hampered by overwhelming confounding by the carcinogenic influence of ultraviolet radiation on skin melanocytes. Nonetheless an inverse association was suggested in a few studies with greater consumption of dairy foods or other dietary sources. Several lines of evidence are consistent with a potential influence for vitamin D on site-specific aggressiveness of skin melanomas, therapeutic response or patient survival. CONCLUSION: Additional research is needed to determine whether vitamin D may have a preventive role in melanoma incidence or a salutary influence on melanoma patient outcome. PMID- 19282201 TI - WITHDRAWN: Intimate partner violence and depression among whites, blacks, and hispanics. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19282202 TI - Gestational diabetic patients with adequate management have normal cardiovascular autonomic regulation during the third trimester of pregnancy and 3 months after delivery. AB - OBJECTIVE: The aim of the present study was to evaluate the influence of gestational diabetes mellitus (GDM) on hemodynamics and cardiovascular autonomic regulation at rest and their responses to head-up tilt (HUT). RESEARCH DESIGN AND METHODS: We prospectively studied 79 pregnant women (51 with GDM, 28 without GDM) during the third trimester of pregnancy and after parturition. The maternal electrocardiogram and arterial blood pressure were noninvasively measured. Heart rate and blood pressure were measured in the supine position and in the upright position. Stroke volume was assessed from noninvasive blood pressure signals, heart rate variability (HRV) was analyzed in frequency domain, and baroreflex sensitivity by the cross-spectral and sequence methods. RESULTS: Between the GDM group and control pregnant women there were no significant differences in hemodynamics and cardiovascular autonomic regulation throughout the protocol. Increased normalized low-frequency component and low-frequency to high-frequency ratio suggested a change in sympathovagal balance towards sympathetic predominance during pregnancy in both groups. The response to head-up tilt (HUT) was similar in both GDM and control pregnant women. The pregnancy modulated the response to HUT in systolic and diastolic blood pressure, stroke volume, cardiac index, peripheral resistance, total power of HRV, and its low- and high-frequency components. CONCLUSIONS: Our results suggest that pregnancy modulates cardiovascular autonomic regulation and hemodynamics equally in subjects with GDM and without GDM, suggesting that metabolic disorder during pregnancy does not result in cardiovascular dysfunction when GDM is in good balance. PMID- 19282203 TI - Factors associated with poor glycemic control among patients with type 2 diabetes. AB - OBJECTIVES: Determine factors associated with poor glycemic control among Jordanian patients with Type 2 diabetes. METHODS: A systematic random sample of 917 patients was selected from all patients with Type 2 diabetes over a period of 6 months in 2008. A prestructured questionnaire sought information about sociodemographic, clinical characteristics, self-care management behaviours, medication adherence, barriers to adherence, and attitude towards diabetes. Weight, height, and waist circumferences were measured. All available last readings of hemoglobin A1c (HbA1c), fasting blood sugar measurements and lipid were abstracted from patients' records. Poor glycemic control was defined as HbA1c >or=7%. RESULTS: Of the total 917 patients, 65.1% had HbA1c >or=7%. In the multivariate analysis, increased duration of diabetes (>7 years vs. C) in the TH gene. l-Dopa therapy in both high and low doses resulted in massive hyperkinesias, while substitution with selegiline exerted only a mild beneficial effect. Today, at the age of 7 years, the patient demonstrates severe developmental retardation with marked trunkal hypotonia, hypokinesia and occasionally dystonic and/or hyperkinetic crises. He is the third Greek patient with TH deficiency to be reported. Since all three patients carry the same pathogenetic mutation, a founder effect is suspected. PMID- 19282211 TI - An unusual cause of a common symptom: pyomyositis presenting with sciatica. PMID- 19282212 TI - [Secularism at hospital]. PMID- 19282213 TI - [The primiparae couple's experience of childbirth: quantitative study]. AB - OBJECTIVE: The aim of this research was to study childbirth experience for the primiparae couple and their emotional state in early post-partum. PATIENTS AND METHODS: Thirty-three couples were interviewed 2 days after the birth. A questionnaire on childbirth experience and the EPDS were carried out. RESULTS: According to the results, mothers feel less satisfied, less calm, more useful and suffer more than fathers. Moreover, they consider father's participation more active and more supportive than themselves assess it. Experience of childbirth is linked with parent's emotional state in early postpartum. DISCUSSION AND CONCLUSION: These results stress the importance to take into account the woman, the men and the couple as three separate and singular units and to create space for dialogue before and after childbirth. PMID- 19282214 TI - [Secularism and State hospital]. PMID- 19282215 TI - The sentence and Kyukei in Japanese criminal procedure, especially in domestic homicide cases. AB - Kyukei is the prosecutor's suggestion of punishment for the accused in the closing argument in the Japanese criminal court. It is rumored that the court sentence is foreseeable in 8 out of 10 of Kyukei. In this study, the ratio of the average sentence against the Kyukei for Oyako-shinju cases was confirmed to be 0.85 for male offenders and 0.71 for female offenders. Although there is a somewhat lenient tendency observed for female offenders, the ratio may almost be found around 0.8. To make a comparative study, we referred to the Kyukei and sentences of out-of-home murder cases and fatal child abuse cases. In the murder case, the ratio of the average sentence against the Kyukei is 0.83, whereas in the child abuse case it is 0.73. What is called the "myth of 8 out of 10" is more applicable for the male offenders who commit the majority of crimes, than for the females. Most of the crimes committed by women are domestic in nature. Indeed, because most women's domestic cases are due to husbands who are absent, behave violently, or are irresponsible, their circumstances of committing the crimes may be taken into account by the judge. PMID- 19282216 TI - Apoptosis and gene expression after TBI. AB - Proper CNS function depends on concerted expression of thousands of genes in a controlled and timely manner. Traumatic brain injury (TBI) in humans results in neuronal death and neurological dysfunction, which might be mediated by altered expression of several genes. By employing a CNS-specific GeneChip and real time polymerase chain reaction (PCR), the study analyzed the gene expression changes. The findings of necrosis and apoptosis can help to improve the wound age estimation after TBI in legal medicine. PMID- 19282217 TI - Immunohistochemical and molecular-pathological diagnosis of myocarditis in cases of suspected sudden infant death syndrome (SIDS)--a multicenter study. AB - Virus-induced myocarditis is a common disease even in infants and children, but diagnosis can be difficult according to the Dallas-criteria, which have been criticised as being too unreliable. The diagnosis has been substantially improved due to immunohistochemical techniques for characterization and quantification of myocardial inflammatory reactions as well as molecular-pathological methods for viral genome detection. The published studies report on post-mortem samples from SIDS victims and controls which were prospectively investigated. Pediatric cases of unnatural deaths served as controls. The results demonstrate a clearly higher prevalence of viral myocardial infections in cases of suspected SIDS. Preliminary criteria for cellular immunohistochemical diagnosis of viral myocardial affections derived from these findings were suggested. PMID- 19282218 TI - Accidental fatal ingestion of colchicine-containing leaves--toxicological and histological findings. AB - A 65 year old woman had collected some plant leaves, cooked them in water and ate this mash with honey. A few hours after consumption first symptoms of intoxication like sickness, stomach aches and diarrhoea appeared. Her condition deteriorated, she was admitted to hospital, became comatose and finally died due to a multiorgan failure. The plants she had collected were identified as autumn crocus (Colchicum autumnale). Colchicine-concentrations detected in blood samples (HPLC/DAD) taken in hospital approximately 24 h after ingestion were 50 microg/l, in post-mortem femoral blood 65 microg/l colchicine were found (toxic range: >24 microg/l). Also all tested organs contained equally high colchicine levels. Histological examination of the liver showed diffuse vacuolization in the cytoplasma of hepatocytes. Possibly due to the relatively short survival time, only isolated mitotic structures were found within the epithelium of the colon. PMID- 19282219 TI - Driving under the influence of drugs and alcohol in Salzburg and Upper Austria during the years 2003-2007. AB - Until 2002 in Austria a blood sample could not be drawn due to regulations stipulated by the Austrian constitution. During the years 2003-2007 alcohol, pharmaceuticals and illicit drugs were analyzed in 1167 blood samples from cases of suspected and drugged drivers. In accordance with the findings of the EU project Rosita, a wide variety of illicit drugs and medications could be found in blood samples of the drivers where cannabis (50%), opiates (20%), amphetamines (18%), cocaine (15%) and benzodiazepines (20%) were those with the highest prevalence. To enable police and medical officers to identify drivers under the influence of cannabis, a newly developed urinary road-site-test system, Check 24 (Protzek GmbH, Germany), with two different cut-off values for THCCOOH glucuronide was used. So far, it was not possible to draw any conclusions from a cannabinoid positive urine sample to the actual influence of a driver due to the previous consumption of cannabis. Using this test a better differentiation between recent and temporal earlier consumption was possible. In addition to using the Check 24 system the technology of pupillography (AMTech GmbH, Germany) was applied in cases of drugged driving. For the first time the authors were able to predict the presence of at least one central nervous active substance in the blood of a drugged driver. PMID- 19282220 TI - Increased expressions and activations of apoptosis-related factors in cell signaling during incised skin wound healing in mice: a preliminary study for forensic wound age estimation. AB - Recent studies have demonstrated that apoptosis plays a pivotal role during skin wound healing and apoptosis-related factors in cell signaling regulate a variety of cellular function. In this study, the expressions of p38MAPK, and JNK, iNOS, eNOS were detected and the activations of caspase-6, -7, -8, -9, and calpain, another signaling pathway of apoptosis, were also investigated by immunohistochemical staining and Western blotting in mice. A time-dependent increase of each protein level was observed by immunohistochemistry and Western blot in mouse skin incision. p38MAPK level peaked at 12 h and 3 d, calpain level peaked at 1 d and 5 d, iNOS level peaked at 1 d and 10 d, while the peak levels of eNOS, caspase-6, -7, -8, and -9 occurred at 3 d and p-JNK at 1 d post-injury. In the early phase of wound healing, infiltrating polymorphonulcear cells were labeled with all the factors except caspase-8. Thereafter, infiltrating mononuclear cells and proliferating spindle-shaped fibroblastic cells showed positive staining for p38MAPK, JNK, calpain, caspases and NOS. The activation of caspase-8, -9, -6, and -7 as detected by Western blot indicated that caspase apoptotic pathway may take effect in cellular elimination during skin wound healing. From the viewpoint of forensic pathology, the time-dependent expressions of the factors in apoptotic pathway during skin incised wound healing may be used as potential markers for wound age estimation. PMID- 19282221 TI - A study of monoamine neuronal systems of schizophrenic patients: using forensic autopsy brains. AB - Studies on autopsy brains of mental illnesses are essential for psychiatric research. Brain materials from forensic autopsy cases have been valuable research resource. We introduced some characteristic neuronal structures of schizophrenic post-mortem brains and normal controls detected using both forensic and pathological autopsy cases and immunohistochemical method. Dopamine-related neuronal structures in schizophrenic cases were investigated by using immunohistochemistry for tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC) and phenylethanolamine-N-methyltransferase (PNMT). At the level of the striatum, the number of AADC-positive, but TH-negative neurons (D neurons) reduced in schizophrenics compared with controls. In the brainstem structures, we described some morphological findings characteristic to catecholaminergic neurons of the ventral tegmental area of schizophrenic cases and reduction of the number of medullary catecholaminergic neurons. These results might be in accordance with recent reports describing neural stem cell dysfunction in schizophrenic brains. The authors also introduced the first systematic psychiatric brain bank in Japan, the Fukushima Psychiatric Brain Bank, constructed in 1997, and mentioned the necessity for arrangement of brain bank systems to further promote the psychiatric brain research. PMID- 19282222 TI - Aggressions against healthcare workers: an approach to the situation in Spain and the victims psychological effects. AB - Aggression against healthcare workers is a problem of important consequences which is becoming a focus of research. However, its possible effects on psychological health have not been studied sufficiently in spite of the fact that they may be of importance even in the absence of physical aggression [Winstnaley S, Whittington R. Aggression towards health care staff in a UK general hospital: variation among professions. J Clin Nurs 2004;13:3-10,[1]]. PMID- 19282223 TI - Pethidine-induced neuronal apoptosis in rat brain. AB - We investigated the effects of pethidine on neurons and the relationship between neuronal apoptosis and pethidine dependence. Thirty male Sprague-Dawley rats were randomly divided into control, dependence and withdrawal group. The animals were treated with saline or pethidine by subcutaneous injection for 60 days. Dose of pethidine was from 20 to 140 mg kg(-1)d(-1). Spontaneous withdrawal was induced by ceasing pethidine administration in withdrawal group. Ultrastructure was observed by transmission electron microscope. The ncNOS, caspase-3 and Bax IHC were performed on paraffin sections by SP method. TUNEL was used as a marker for identification of neuronal apoptosis in cerebral cortex and periaqueductal grey matter. Under ultrastructure, there were many features of neuronal apoptosis and necrosis. There also were many TUNEL staining positive neurons scattered in brain tissue. The grey scale of IHC staining and number of positive neurons of ncNOS, caspase-3 and Bax increased distinctly in chronic dependence and withdrawal (P<0.05). Pethidine resulted in neuronal apoptosis, degeneration and necrosis. These damages could be pathological basis of pethidine dependence and tolerance. PMID- 19282224 TI - Awareness in nine countries: a public health approach to suicide prevention. AB - Suicide is an important public health problem, increasing worldwide, and on a yearly basis accounting for the death of more than one million people, with estimates as high as 10-20 times that many attempting to take their own life. Because successful suicide prevention depends upon recognition of symptoms of mental ill-health, awareness of these signs is a necessary precondition. The ability and responsibility for recognizing signs and symptoms of suicide, until most recently, however, was the exclusive purview of mental health professionals. Lately, there have been efforts to screen high risk populations and to train others to effectively respond to suicidal behavior, including classic first responders, primary care providers, hot line operators, teachers, etc. But what about everyone else who may have an opportunity to prevent a suicide simply by knowing when to ask questions, what to listen for, and understanding when additional assistance is warranted? What about the suicidal person who wants to tell someone about their distress but "knows" that such a conversation will not help nor be well-received? Where does a person living where mental health services are lacking or are beyond one's financial means turn to for relief and assistance? Does not Public Health have something to offer in response to these pressing questions? In 2002-2005, a study was carried out in nine countries, distributed over five continents, under the auspices and support of the Presidential Commission of the World Psychiatric Association (WPA), the World Health Organization (WHO), and the International Association of Child and Adolescent Psychiatry and Allied Professions (IACAPAP), to test the feasibility and effectiveness of raising awareness and increasing knowledge about child mental health, including suicidality, among students, teachers and parents. Implications for this approach as a model for suicide prevention are presented. PMID- 19282225 TI - Characteristics of chest injury in falls from heights. AB - This study included 660 victims injured due to high fall. All cases are with injuries caused by the free-fall model of high fall. The range of the heights was from 0 to 70 m. The sample was divided into three groups of growing height: low falls up to 7 m, high falls from 7 to 30 m, and very high falls above 30 m. Statistical analysis was performed using two way ANOVA according to Scheirer-Ray Hare in complete uneven block. Males were 71% (469), females 29% (191). In the analyzed sample, apart from the height of fall, the number of accidents is about 44% (290 cases) and suicides about 56% (370 cases). Thirty six percent, i.e., 238 of 660 victims had consumed alcohol before death (alcoholemia >0.5 per thousand). The objective of our study was to analyze characteristics of chest injuries made by falling from a height. The frequency of thoracic injuries is statistically significantly related to the height of fall (p<0.05); it increases constantly in falls from heights beyond 3m, so that the rib fractures were present in all falls from heights beyond 40m. The aortic ruptures were found in 21%, and the heart injuries in 16% of all cases. The frequency of aortic and heart ruptures was not statistically significantly related to the height of fall (p>0.05). A rupture of the aorta was the most commonly located on its arch (82%). The heart ruptures were the most commonly located on the atrial posterior wall (70%). PMID- 19282226 TI - Sudden death, especially in infancy--improvement of diagnoses by biochemistry, immunohistochemistry and molecular pathology. AB - One of the problems in the diagnosis of the cause of death in cases of sudden death, especially in infancy, is the rapidity of death and that the morphological correlates of the underlying diseases and cause of death may be scarce or even completely missing. This is especially true for functional disorders causing death (e.g. arrhythmias) or cases where death occurs in an initial stage of disease with still lacking morphological findings (e.g. myocarditis). Molecular pathological techniques, which were initially of great importance for identification, today contribute also to the determination of the cause and manner of death, especially in cases, where classical methods do not reveal a clear anatomical cause of death. This will be addressed on the basis of several case groups, especially cases of sudden infant death syndrome (SIDS). Using immunohistochemical methods with qualification and quantification of interstitial leucocytes, PCR and Rt-PCR methods for identifying virus genome within the myocardium, it is possible to identify in about 25% of SIDS cases a myocarditis as cause of death. However, proposed limit values for the diagnosis of myocarditis have to be seen with caution since they lack any statistical power. The value of immunohistochemical and molecular pathological methods to identify the cause of death will also be addressed in cases of sudden death of young adults. At last pharmacogenomic investigations, e.g. on the metabolism of tramadol will be addressed which are of importance to declare adverse events, or even lethal outcome during medication. PMID- 19282227 TI - Forensic DNA typing in China. AB - In the field of forensic genetics, essential developmental impulses come from the advances of the molecular biology and human genome projects. This paper overviews existing technologies for forensic genetics in China and gives a perspective of forensic DNA analysis. In China, work has been done in the development of blood group serology of the conventional markers. Forensic scientists in China also contributed to the progress of DNA analysis by the validation of numerous test methods and by optimization of these methods. During these years, forensic DNA analysis in China has experienced tremendous progress towards development of robust, efficient and precise protocols, including the development of short tandem repeat analysis, mitochondrial DNA and Y-chromosome analysis. Forensic scientists are constantly looking for new methods to further improve DNA typing. Therefore, this paper also focuses on emerging new technologies in China, which represent an interest for forensic genetics. PMID- 19282228 TI - Suicide by sharp force injuries--a study in Oporto. AB - A study was undertaken on suicidal cases due to sharp force injuries (cut and/or stab wounds) observed at the Northern Branch of the National Institute of Legal Medicine (Portugal) from 1997 to 2007. Its aim was to analyse the different wound patterns and the epidemiological and social factors that could be related to the suicide behaviour in these subjects, through the examination of autopsy records and information from the social inquiry. A total of 16 cases were found, with an age range from 19 to 79 years old (mean=55.6 yrs) and a male predominance (M=10; F=6). Symptoms of depression (or the clinical diagnosis itself) had been reported in 5 cases. A total of nine individuals had previously expressed some sort of suicidal ideation and eight had already attempted suicide. A kitchen knife was the most commonly used object (n=8). Fatal incised wounds were prominent (n=13) and the neck was the anatomical region more often fatally wounded (n=6). Tentative wounds, which are non-fatal, more superficial, cut or stab wounds, were found in 8 cases. Hesitation marks were observed in close proximity of the fatal wound in 6 cases. PMID- 19282229 TI - Biostatistical study of suicide features in Mures County (Romania). AB - Mures County (580,000 inhabitants) has constantly higher yearly suicide rates (23 32%ooo) compared to Romania's national average (12-14%ooo). We performed a prospective study with the aim to find characteristics that should help the preventive management. Autopsy is compulsory by law in violent deaths; therefore, all suicide cases are referred to our Institute. During the period 2005-2006 we completed a multi-item questionnaire in each suicide case from a total of 273. Beside age, sex, marital status, occupation and economical status, we also investigated religious adherence, type of alcoholic beverages consumed, blood alcohol concentration, previous suicide attempts, farewell notes, medical background, behavioral changes. RESULTS: sex distribution was M:F=4.8:1; average age for suicide victims was 51 years, the interquartile range was 38-62 years for men and 45-70 years for women. Hanging is by far the most used method for suicide in our region, 80% of the cases. Marital status (married/non-married) does not seem to influence suicide conduct. Social inactivity (workless, non-occupied) was observed in 65% of the victims. In rural areas we found a double suicide rate, rural/urban ratio was 2/1 while the population is equally distributed in towns and villages in our province (50%); we also noticed a seasonal variation of suicides in rural areas (with peaks in June and August) whereas in urban areas differences were statistically not significant. Ethnic Hungarians have double suicide rates compared to Romanians. Recent alcohol ingestion was noticed in 60% of men and only 27% of women suicides. PMID- 19282230 TI - The proof of flat-line scalp EEGs of brain dead patients by an automatic EEG analysis system. AB - Scalp electroencephalograms (EEGs) of brain dead patients are macroscopically flat under 7 or 10 microV/mm electroencephalograph sensitivity, but significant noises are detected in EEGs under 2 microV/mm sensitivity, interfering with the analysis. EEGs of 20 brain dead patients (17-76 years old) were therefore analyzed quantitatively as equivalent electric potentials in frequency bands delta, theta, alpha and beta using the automatic EEG analysis system developed in Kansai Medical University. The equivalent electric potentials in each band were about or less than 1 microV, which is used as a criterion of judgment of flat line EEGs or brain death. Then, macroscopically flat EEGs of 12 comatose patients including infants (3-67 years old) were analyzed by the system, confirming their brain death. Thus, the automatic EEG analysis system could be used as a supporting tool to confirm flat-line EEGs of brain dead patients. ATAMAP II for Windows software was also evaluated. PMID- 19282231 TI - Inactivation of Escherichia coli and Streptococcus mutans by ultrasound at 500kHz. AB - This paper shows a systematic study of the 500 kHz frequency ultrasound efficiency on the microbial inactivation as a function of ultrasonic power delivered into the bacterial suspension. The inactivation of Escherichia coli IAM 12058, a Gram-negative bacterium and Streptococcus mutans JCM 5175, a Gram positive bacterium is enhanced by increasing the ultrasonic power in the range of 1.7-12.4W and the logarithm of survival ratio decreases linearly with irradiation time, except for E. coli sonicated with the highest power level. The rate constants were estimated in the linear region of the plots representing survival ratio logarithm vs. sonication time. A better understanding of the inactivation process at 500kHz could be gained by suppressing the chemical effects with a radical scavenger. We find out that the rate constants increase with the ultrasonic power delivered into the solution and dramatically decrease by the addition of t-butanol as a radical scavenger to the bacterial suspension. For comparison, experiments were carried out at a low frequency level of 20kHz. It was found out that for the same ultrasonic power delivered into the bacterial suspension, the inactivation was slightly enhanced at 500kHz frequency. The examinations of bacterium performed with a TEM revealed lethal damages arising from the interaction of bacterial cells with the cavitational bubbles. A significant amount of empty cell envelopes as well as their cytoplasmatic content was detected. Thus, based on these new data, the mechanism of bacterial inactivation by ultrasounds at high frequency is discussed here. PMID- 19282232 TI - Sonophotocatalysis in advanced oxidation process: a short review. AB - Sonophotocatalysis involves the use of a combination of ultrasonic sound waves, ultraviolet radiation and a semiconductor photocatalyst to enhance a chemical reaction by the formation of free radicals in aqueous systems. Researchers have used sonophotocatalysis in a variety of investigations i.e. from water decontamination to direct pollutant degradation. This degradation process provides an excellent opportunity to reduce reaction time and the amount of reagents used without the need for extreme physical conditions. Given its advantages, the sonophotocatalysis process has a futuristic application from an engineering and fundamental aspect in commercial applications. A detailed search of published reports was done and analyzed in this paper with respect to sonication, photocatalysis and advanced oxidation processes. PMID- 19282233 TI - Unsupervised decomposition of low-intensity low-dimensional multi-spectral fluorescent images for tumour demarcation. AB - Unsupervised decomposition of static linear mixture model (SLMM) with ill conditioned basis matrix and statistically dependent sources is considered. Such situation arises when low-dimensional low-intensity multi-spectral image of the tumour in the early stage of development is represented by the SLMM, wherein tumour is spectrally similar to the surrounding tissue. The original contribution of this paper is in proposing an algorithm for unsupervised decomposition of low dimensional multi-spectral image for high-contrast tumour visualisation. It combines nonlinear band generation (NBG) and dependent component analysis (DCA) that itself combines linear pre-processing transform and independent component analysis (ICA). NBG is necessary to improve conditioning of the extended mixing matrix in the SLMM, while DCA is necessary to increase statistical independence between spectrally similar sources. We demonstrate good performance of the method on both computational model and experimental low-intensity red-green-blue fluorescent image of the surface tumour (basal cell carcinoma). We believe that presented method can be of use in other multi-channel medical imaging systems. PMID- 19282234 TI - An augmented reality system for liver thermal ablation: design and evaluation on clinical cases. AB - We present in this paper an augmented reality guidance system for liver thermal ablation in interventional radiology. To show the relevance of our methodology, the system is incrementally evaluated on an abdominal phantom and then on patients in the operating room. The system registers in a common coordinate system a preoperative image of the patient and the position of the needle that the practitioner manipulates. The breathing motion uncertainty is taken into account with a respiratory gating technique: the preoperative image and the guidance step are synchronized on expiratory phases. In order to fulfil the real time constraints, we have developed and validated algorithms that automatically process and extract feature points. Since the guidance interface is also a major component of the system effectiveness, we validate the overall targeting accuracy on an abdominal phantom. This experiment showed that a practitioner can reach a predefined target with an accuracy of 2mm with an insertion time below one minute. Finally, we propose a passive evaluation protocol of the overall system in the operating room during five interventions on patients. These experiments show that the system can provide a guidance information during expiratory phases with an error below 5mm. PMID- 19282235 TI - Modeling QTL for complex traits: detection and context for plant breeding. AB - The genetic architecture of a trait is defined by the set of genes contributing to genetic variation within a reference population of genotypes together with information on their location in the genome and the effects of their alleles on traits, including intra-locus and inter-locus interactions, environmental dependencies, and pleiotropy. Accumulated evidence from trait mapping studies emphasizes that plant breeders work within a trait genetic complexity continuum. Some traits show a relatively simple genetic architecture while others, such as grain yield, have a complex architecture. An important advance is that we now have empirical genetic models of trait genetic architecture obtained from mapping studies (multi-QTL models including various genetic effects that may vary in relation to environmental factors) to ground theoretical investigations on the merits of alternative breeding strategies. Such theoretical studies indicate that as the genetic complexity of traits increases the opportunities for realizing benefits from molecular enhanced breeding strategies increase. To realize these potential benefits and enable the plant breeder to increase rate of genetic gain for complex traits it is anticipated that the empirical genetic models of trait genetic architecture used for predicting trait variation will need to incorporate the effects of genetic interactions and be interpreted within a genotype environment-management framework for the target agricultural production system. PMID- 19282236 TI - Damage control: regulating defenses against toxic metals and metalloids. AB - Some elements are essential for life and others closely related to them are very toxic. In exploiting unique ecological niches many prokaryotes have evolved the means to defend themselves against and even to derive energy from deleterious elements. Toxic metal defense systems are related to those providing homeostasis of essential metals and metalloid elements. Expression of these multiprotein systems is costly but they must respond rapidly and, so, all are well controlled. Seven diverse families of metalloregulators are presently recognized for essential metal homeostasis in prokaryotes. Two of these, the ArsR and MerR families, figure more often than the others in controlling responses to toxic transition metals and metalloids. This review emphasizes recent advances in these two metalloregulator families and highlights emerging regulatory motifs of other types. PMID- 19282237 TI - A pilot study of sleep, cognition, and respiration under 4 weeks of intermittent nocturnal hypoxia in adult humans. AB - STUDY OBJECTIVES: A pilot study to examine the effects of intermittent nocturnal hypoxia on sleep, respiration and cognition in healthy adult humans. METHODS: Participants were eight healthy, non-smoking subjects (four male, four female), mean age of 26.4+/-5.2 years, and BMI 22.3+/-2.6 kg/m(2), exposed to 9h of intermittent hypoxia between the hours of 10 P.M. and 7 A.M. for 28 consecutive nights. At a simulated altitude of 13,000 feet (FIO(2) 0.13), intermittent hypoxia was achieved by administering nasal nitrogen, alternating with brief (approximately 5s) boluses of nasal oxygen. Pre- and post-exposure assessments included polysomnography, attention (20-min Psychomotor Vigilance Test), working memory (10-min verbal 2 and 3-back), Multiple Sleep Latency Test, and the Rey Auditory Verbal Learning Test. Obstructive and non-obstructive respiratory events were scored. RESULTS: Overall sleep quality showed worsening trends but no statistically significant change following exposure. There was no difference after hypoxia in sleepiness, encoding, attention or working memory. Hyperoxic central apneas and post-hyperoxic respiratory instability were noted as special features of disturbed respiratory control induced by intermittent nocturnal hypoxia. CONCLUSIONS: In this model, exposure to nocturnal intermittent hypoxia for 4 weeks caused no significant deficits in subjective or objective alertness, vigilance, or working memory. PMID- 19282238 TI - Multigene evidence for the placement of a heterotrophic amoeboid lineage Leukarachnion sp. among photosynthetic stramenopiles. AB - The colorless amoeboid eukaryote genus Leukarachnion represents one of a long list of microbial lineages for which there have been few taxonomic studies. In this study, we analyze molecular data to assess the placement of a species of Leukarachnion on the eukaryotic tree of life and we report fine structural data to provide additional information on the identity of this taxon. Our multigene analyses indicate that Leukarachnion sp. (ATCC PRA-24) is a member of the stramenopiles, sister to the Chrysophyceae/Synurophyceae clade. It also forms a sister group relationship to the clade containing Chlamydomyxa labyrinthuloides and Synchroma grande, both of which are characterized by net-like amoeboid phases. Leukarachnion sp. and Chlamydomyxa labyrinthuloides also share fine structural cyst morphology such as bilayered structure of the cyst wall. The amoeboid form and heterotrophic habit of Leukarachnion sp. highlight the multiple origins of diverse body forms and multiple plastid losses within the stramenopiles. PMID- 19282239 TI - Hope as experienced in women newly diagnosed with gynaecological cancer. AB - AIM: This article presents findings from a hermeneutic-phenomenological study with the aim to investigate the meaning of the lived experience of hope in women newly diagnosed with gynaecological cancer. METHOD: Fifteen women were interviewed the day they were receiving the diagnosis at a gynaecological department of a Danish university hospital. The women, aged 24-87 (median 52 yrs), were diagnosed with ovarian, endometrial, cervical and vulvar cancer. RESULTS: Hope was found to be connected to both diagnosis, cure, family life and life itself and closely tied to hopelessness. The newly received cancer diagnosis made the women oscillate between hope and hopelessness, between positive expectations of getting cured and frightening feelings of the disease taking over. Five major interrelated themes of hope were identified: hope of being cured, cared for and getting back to normal, hope as being active and feeling well, hope as an internal power to maintain integration, hope as significant relationships and hope as fighting against hopelessness. Thus, hope was woven together with hopelessness in a mysterious way; it took command through inner strength and courage based on a trust in being cured and of being in relationship with significant others. CONCLUSION: The findings of the close relationship between the shades of hope and hopelessness support the need for nurses to continue to practice hope-inspiring nursing. Nurses need to understand the complexity of hope and its close connection to hopelessness when newly diagnosed with a threatening disease as cancer; and the findings might help nurses assist patients in fighting hopelessness. PMID- 19282240 TI - The meaning of cancer for Australian Aboriginal women; changing the focus of cancer nursing. AB - AIM: The purpose of the study was to explore why Aboriginal women participate in cancer screening programs but appear reluctant to following-up results, or accept medical advice about treatment. METHODS: Interpretive ethnography, a qualitative methodology, was used to explore Aboriginal women's perception of cancer, and the cultural context in which meaning was constructed and influenced treatment decision. Data collection, which occurred over two years, involved fieldwork, participant-observation, face-to-face interviews and focus groups, in two rural Aboriginal communities. Forty eight interviews were recorded from a cross section of the communities, including cancer survivors and patients, family members, health care providers and other women from the community. RESULTS: Key findings were that Aboriginal women's had a fearful and fatalistic attitude toward cancer, doubted the efficacy of treatment and carried an enduring ambivalence toward the authority of whiteman's medicine. The women faced a dilemma of wanting access to cancer treatment options but feared entering hospital or clinics not attuned to their cultural needs. CONCLUSION: The findings highlight the need for a culture centred approach that decentres the authority of conventional services and instead gives prominence to Aboriginal cultural values as a focal point in cancer control. It should be the responsibility of cancer nurses and others to engage with their local Aboriginal communities to build relationships that foster an exchange of learning about cultural differences that make a difference to how cancer control is practiced. PMID- 19282241 TI - Sensitivity and specificity of multimodal and ultrasound screening for ovarian cancer, and stage distribution of detected cancers: results of the prevalence screen of the UK Collaborative Trial of Ovarian Cancer Screening (UKCTOCS). AB - BACKGROUND: Ovarian cancer has a high case-fatality ratio, with most women not diagnosed until the disease is in its advanced stages. The United Kingdom Collaborative Trial of Ovarian Cancer Screening (UKCTOCS) is a randomised controlled trial designed to assess the effect of screening on mortality. This report summarises the outcome of the prevalence (initial) screen in UKCTOCS. METHODS: Between 2001 and 2005, a total of 202 638 post-menopausal women aged 50 74 years were randomly assigned to no treatment (control; n=101 359); annual CA125 screening (interpreted using a risk of ovarian cancer algorithm) with transvaginal ultrasound scan as a second-line test (multimodal screening [MMS]; n=50 640); or annual screening with transvaginal ultrasound (USS; n=50 639) alone in a 2:1:1 ratio using a computer-generated random number algorithm. All women provided a blood sample at recruitment. Women randomised to the MMS group had their blood tested for CA125 and those randomised to the USS group were sent an appointment to attend for a transvaginal scan. Women with abnormal screens had repeat tests. Women with persistent abnormality on repeat screens underwent clinical evaluation and, where appropriate, surgery. This trial is registered as ISRCTN22488978 and with ClinicalTrials.gov, number NCT00058032. FINDINGS: In the prevalence screen, 50 078 (98.9%) women underwent MMS, and 48 230 (95.2%) underwent USS. The main reasons for withdrawal were death (two MMS, 28 USS), non ovarian cancer or other disease (none MMS, 66 USS), removal of ovaries (five MMS, 29 USS), relocation (none MMS, 39 USS), failure to attend three appointments for the screen (72 MMS, 757 USS), and participant changing their mind (483 MMS, 1490 USS). Overall, 4355 of 50 078 (8.7%) women in the MMS group and 5779 of 48 230 (12.0%) women in the USS group required a repeat test, and 167 (0.3%) women in the MMS group and 1894 (3.9%) women in the USS group required clinical evaluation. 97 of 50 078 (0.2%) women from the MMS group and 845 of 48 230 (1.8%) from the USS group underwent surgery. 42 (MMS) and 45 (USS) primary ovarian and tubal cancers were detected, including 28 borderline tumours (eight MMS, 20 USS). 28 (16 MMS, 12 USS) of 58 (48.3%; 95% CI 35.0-61.8) of the invasive cancers were stage I/II, with no difference (p=0.396) in stage distribution between the groups. A further 13 (five MMS, eight USS) women developed primary ovarian cancer during the year after the screen. The sensitivity, specificity, and positive predictive values for all primary ovarian and tubal cancers were 89.4%, 99.8%, and 43.3% for MMS, and 84.9%, 98.2%, and 5.3% for USS, respectively. For primary invasive epithelial ovarian and tubal cancers, the sensitivity, specificity, and positive-predictive values were 89.5%, 99.8%, and 35.1% for MMS, and 75.0%, 98.2%, and 2.8% for USS, respectively. There was a significant difference in specificity (p<0.0001) but not sensitivity between the two screening groups for both primary ovarian and tubal cancers as well as primary epithelial invasive ovarian and tubal cancers. INTERPRETATION: The sensitivity of the MMS and USS screening strategies is encouraging. Specificity was higher in the MMS than in the USS group, resulting in lower rates of repeat testing and surgery. This in part reflects the high prevalence of benign adnexal abnormalities and the more frequent detection of borderline tumours in the USS group. The prevalence screen has established that the screening strategies are feasible. The results of ongoing screening are awaited so that the effect of screening on mortality can be determined. PMID- 19282242 TI - CD154 and its receptors in inflammatory vascular pathologies. AB - CD154, a member of the tumor necrosis factor-alpha family, has recently been implicated in the pathophysiology of vascular diseases. Indeed, blockade of CD154 by neutralizing antibodies or genetic disruption in mice prevents atherosclerosis and atherothrombosis. CD154 is believed to interact mainly via the CD40 receptor, however, it has also been found to bind with alphaIIbbeta3 integrin and so induces platelet activation. Moreover, we (and others) have recently identified the integrins alpha5beta1 and Mac-1 as novel CD154 receptors expressed on many cell types. Here, we illustrate the various functional features of these molecules, while describing the increasingly important role of CD40 in CD154 associated vascular pathologies such as atherosclerosis and atherothrombosis. PMID- 19282243 TI - NK cell education: not an on-off switch but a tunable rheostat. AB - Heterogeneity in the natural killer (NK) cell population is determined by variegated expression of polygenic and polymorphic receptors and distinct phenotypes and functions of NK cell subsets. Here, we outline an additional heterogeneity at the level of MHC-dependent education of NK cells. Based on data obtained using polychromatic flow cytometry and mice with single MHC class I alleles, we argue that NK cell responsiveness is tuned along a continuum determined by the strength of the inhibitory input received by the individual NK cell during education. This model has implications for the use of NK cells in therapeutic settings and affects interpretations of how NK cells control virus infections and regulate autoimmunity. PMID- 19282244 TI - Germinal centres seen through the mathematical eye: B-cell models on the catwalk. AB - Germinal centres are receiving renewed attention following recent intravital multi-photon imaging studies. These data have shed new light on longstanding questions about the spatial organisation of germinal centres, B-cell migration, selection and differentiation. Mathematical models have proven invaluable in the analysis of intravital motility data, and have predicted novel B-cell selection mechanisms that are now supported by experimental findings. We argue that mathematical modelling adds a different vantage point to experimental data and provides a quantitative and systematic analysis of hypotheses and theories in immunology. Furthermore, the well-characterised nature of the germinal centre provides an excellent proving ground for mathematical modelling. PMID- 19282245 TI - Person-centred care and care mapping in dementia. PMID- 19282246 TI - Caring for Aged Dementia Care Resident Study (CADRES) of person-centred care, dementia-care mapping, and usual care in dementia: a cluster-randomised trial. AB - BACKGROUND: Evidence for improved outcomes for people with dementia through provision of person-centred care and dementia-care mapping is largely observational. We aimed to do a large, randomised comparison of person-centred care, dementia-care mapping, and usual care. METHODS: In a cluster randomised controlled trial, urban residential sites were randomly assigned to person centred care, dementia-care mapping, or usual care. Carers received training and support in either intervention or continued usual care. Treatment allocation was masked to assessors. The primary outcome was agitation measured with the Cohen Mansfield agitation inventory (CMAI). Secondary outcomes included psychiatric symptoms including hallucinations, neuropsychological status, quality of life, falls, and cost of treatment. Outcome measures were assessed before and directly after 4 months of intervention, and at 4 months of follow-up. Hierarchical linear models were used to test treatment and time effects. Analysis was by intention to treat. This trial is registered with the Australia and New Zealand Clinical Trials Registry, number ACTRN12608000084381. FINDINGS: 15 care sites with 289 residents were randomly assigned. Pairwise contrasts revealed that at follow-up, and relative to usual care, CMAI score was lower in sites providing mapping (mean difference 10.9, 95% CI 0.7-21.1; p=0.04) and person-centred care (13.6, 3.3 23.9; p=0.01). Compared with usual care, fewer falls were recorded in sites that used mapping (0.24, 0.08-0.40; p=0.02) but there were more falls with person centred care (0.15, 0.02-0.28; p=0.03). There were no other significant effects. INTERPRETATION: Person-centred care and dementia-care mapping both seem to reduce agitation in people with dementia in residential care. PMID- 19282247 TI - Dating and growth in the first trimester. AB - Measurement of embryonic or foetal size using the greatest length of the embryo or foetal crown rump length can be used to accurately determine the gestational age of a normal first trimester pregnancy to within three to five days. Transvaginal ultrasound scan can be used to measure the size of an embryo and gestation sac earlier than transabdominal ultrasound. The original Robinson curve used for dating pregnancies is still valid in most cases. Ultrasound dating in the first trimester is now recommended for all women with spontaneous pregnancies, even those with certain menstrual dates. First trimester growth in normal pregnancy is not uniform and is influenced by both maternal and foetal factors. Early foetal growth restriction is demonstrated in many pregnancies that subsequently end in first trimester miscarriage and is also demonstrated in fetuses with triploidy, trisomy 18 and possibly trisomy 13. Pregnancies which are small at the 11-14 week ultrasound scan appear to be at risk of later intrauterine growth restriction, preeclampsia and preterm delivery. Cross sectional and serial measurement of foetal growth in the first trimester may be helpful in predicting both miscarriage and adverse late pregnancy outcomes. PMID- 19282248 TI - Voltammetric and spectral characterization of two flavonols for assay-dependent antioxidant capacity. AB - Polyphenolic compounds usually showed different antioxidant capacities depending on the assay methods used. To determine the possible chemical cause for this assay-dependence, two flavonols, kaempferol and morin, were selected as the model molecules for the comparative investigation between electrochemical and chemical oxidations. The electro-oxidation of the flavonols was studied using cyclic voltammetry and in situ UV-Vis spectroelectrochemical technique with a long optical-path thin-layer electrolytic cell. The spectral changes recorded in different potentials were compared with those in the chemical oxidation by H(2)O(2) or ABTS(*+) radical in the same thin-layer cell. The 4'-OH group of either sample was first oxidized at lower potentials or induced by H(2)O(2), and in this case kaempferol was somewhat more active than morin. With an additional 2'-OH group, morin underwent the secondary oxidation in moderately higher potentials or by ABTS(*+), showing antioxidant capacity about twice of that of kaempferol. This study clarified that the chemical oxidation of a polyphenolic compound by the oxidants with different reactivities, which corresponded to its electro-oxidation in different anodic peaks, had a difference in number of oxidizable OH-groups, leading to the difference in antioxidant capacity. PMID- 19282249 TI - Dielectric dispersion of suspended cells using 3D reconstructed morphology model. AB - In the framework of this study, novel method for dispersion analysis of cellular suspensions is presented. The method is fundamentally based on the ability to reconstruct the exact 3D morphology of a given cell with resolution accuracy of few nanometers using AFM imaging. By applying a reverse engineering approach, the morphology of the cell is constructed based on a set of measured spatial points that describes its geometry. The permittivity spectrum of the reconstructed cell is then directly derived based on computational solution of complex potential problem using 3D Boundary Element Method. The applicability of the method is demonstrated both theoretically and experimentally with tight comparison to the well known shell models. This comparison reveals significant deviations between the models, and hence, emphasises the vast effect of morphology in dispersion analysis of cellular suspensions. PMID- 19282250 TI - WITHDRAWN: Clinical development of compactin (CS-500). AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19282251 TI - Reciprocal regulation of nuclear import of the yeast MutSalpha DNA mismatch repair proteins Msh2 and Msh6. AB - DNA mismatch recognition is performed in eukaryotes by two heterodimers known as MutSalpha (Msh2/Msh6) and MutSbeta (Msh2/Msh3) that must reside in the nucleus to function. Two putative Msh2 nuclear localization sequences (NLS) were characterized by fusion to green fluorescent protein (GFP) and site-directed mutagenesis in the context of Msh2. One NLS functioned in GFP targeting assays and both acted redundantly within Msh2. We examined nuclear localization of each of the MutS monomers in the presence and absence of their partners. Msh2 translocated to the nucleus in cells lacking Msh3 and Msh6; however, cells lacking Msh6 showed significantly decreased levels of nuclear Msh2. Furthermore, the overall protein levels of Msh2 were significantly diminished in the absence of Msh6, particularly if Msh2 lacked a functional NLS. Msh3 localized in the absence of Msh2, but Msh6 localization depended on Msh2 expressing functional NLSs. Overall, the nuclear levels of Msh2 and Msh6 decline when the other partner is absent. The data suggest a stabilization mechanism to prevent free monomer accumulation in the cytoplasm. PMID- 19282252 TI - Synthesis of the flavonoid 3',4',5'-trimethoxyflavonol and its determination in plasma and tissues of mice by HPLC with fluorescence detection. AB - 3',4',5'-Trimethoxyflavonol (TMFol) was synthesized as a potential colorectal cancer chemopreventive agent. An HPLC method for determination for TMFol in murine plasma and tissues was developed and validated using human plasma. Analyte was separated (C(18) column; fluorescence detection 330nm excitation, 440nm emission) using 69% methanol and 0.1M ammonium acetate buffer (pH 5.1) as mobile phase. The method was linear for 50-2500ng/ml plasma and 0.05-10microg/g tissue (r>0.99). TMFol was recovered from plasma or tissues using solid phase columns or organic solvent protein precipitation, respectively. Recovery at low, medium and high concentrations was 97.6-107.3%, with inter- and intra-day coefficients of variation of <10%. The lower limit of quantitation for plasma was 50ng/ml. The method was applied to measure steady-state TMFol plasma and tissue levels in mice which received dietary TMFol (0.2%). PMID- 19282253 TI - Sensitive and cost-effective LC-MS/MS method for quantitation of CVT-6883 in human urine using sodium dodecylbenzenesulfonate additive to eliminate adsorptive losses. AB - CVT-6883, a novel selective A(2B) adenosine receptor antagonist currently under clinical development, is highly lipophilic and exhibits high affinity for non specific binding to container surfaces, resulting in very low recovery in urine assays. Our study showed the use of sodium dodecylbenzenesulfonate (SDBS), a low cost additive, eliminated non-specific binding problems in the analysis of CVT 6883 in human urine without compromising sensitivity. A new sensitive and selective LC-MS/MS method for quantitation of CVT-6883 in the range of 0.200 80.0ng/mL using SDBS additive was therefore developed and validated for the analysis of human urine samples. The recoveries during sample collection, handling and extraction for the analyte and internal standard (d(5)-CVT-6883) were higher than 87%. CVT-6883 was found stable under the following conditions: in extract - at ambient temperature for 3 days, under refrigeration (5 degrees C) for 6 days; in human urine (containing 4mM SDBS) - after three freeze/thaw cycles, at ambient temperature for 26h, under refrigeration (5 degrees C) for 94h, and in a freezer set to -20 degrees C for at least 2 months. The results demonstrated that the validated method is sufficiently sensitive, specific, and cost-effective for the analysis of CVT-6883 in human urine and will provide a powerful tool to support the clinical programs for CVT-6883. PMID- 19282254 TI - A modified extraction protocol enables detection and quantification of celiac disease-related gluten proteins from wheat. AB - The detection, analysis, and quantification of individual celiac disease (CD) immune responsive gluten proteins in wheat and related cereals (barley, rye) require an adequate and reliable extraction protocol. Because different types of gluten proteins behave differently in terms of solubility, currently different extraction protocols exist. The performance of various documented gluten extraction protocols is evaluated for specificity and completeness by gel electrophoresis (SDS-PAGE), immunoblotting and RIDASCREEN Gliadin competitive ELISA. Based on these results, an optimized, two-step extraction protocol has been developed. PMID- 19282255 TI - [Assisted reproduction techniques and the health of the newborn]. PMID- 19282256 TI - [Birth characteristics due to in vitro fertilization (IVF) techniques]. AB - OBJECTIVE: To determine the number of deliveries after 31 weeks gestation (w), from in vitro fertilization (IVF), including Intracytoplasmic Sperm Injection (ICSI), and to compare characteristics with those achieved by natural fertilization (NF). We analyse twin and singleton pregnancies separately. PATIENTS AND METHOD: This descriptive and prospective study monitors children from foetus, from 12 weeks after IVF, to 12 months of age, including the newborn period. There were 7008 newborns between 1st January 2004 and 31st March 2007 of which there were 113 from IVF and 6895 from NF. RESULTS: Twin-pregnancies in NF is 2.4%, whereas in IVF it is 56.6%, (p<0.001). Mean mother age is 28+/-9 years (y) in NF and 36+/-4y in IVF singletons (p<0.05), and 29+/-10 y in NF and 35+/-4 y in IVF twins (p<0.05). Mean gestational age is 38+/-4 w and 39+/-2w in NF and IVF singletons and 36+/-1 w in NF and IVF twins. No statistically significant differences were found in prematurity rate (<37 w) and low birth weight (< 2500g) between singletons and twins from IVF or NF. Caesarean sections were 23.13% and 51.02% from singleton NF and IVF (p<0.001). No statistically differences were found between twins NF (71.42%) and IVF (79.69%). Malformations at birth were observed in 5.3% of IVF and in 1.1% of NF newborns (p<0.002). Relative risk of malformations (RR) IVF versus NF is 4.83 (95% CI, 2.14-10.83). CONCLUSIONS: Only 1.61% of newborns comes from IVF techniques. There are significant differences in age of mother, twin- pregnancies, caesarean sections (singletons) and congenital defects between them and NF newborns. There were no differences found in prematurity and low birth weight between them and NF newborns, when compared by number of foetus. PMID- 19282257 TI - [Cocaine-induced acute myocardial infarction]. PMID- 19282258 TI - [Treatment of Cushing's disease with ketoconazole]. AB - Cushing's disease (CD), which is caused fundamentally by a pituitary microadenoma, is a rare endocrine disease in the paediatric age and which is associated with significant morbidity. The current treatment of choice for CD is trans-sphenoidal selective adenomectomy resulting in long-term remission rates. Pharmacological management of CD is directed at decreasing adrenal steroid secretion. Ketoconazole inhibits adrenal enzyme activity and decreases cortisol secretion. Our patient is a girl of 13 years and eight months of age with CD. The clinical manifestations were obesity, decrease in growth rate, Cushingoid phenotype, headache and arterial hypertension. The hormone determinations showed elevated values of UFC and ACTH. The circadian rhythm cortisol and ACTH and the short and long dexamethasone tests of provide definitive diagnosis of CD. The cranial MRI with gadolinium suggests the presence of microadenoma in the right pituitary lobe. The diagnosis is confirmed after performing a selective catheterisation of the inferior petrosal sinuses with CRH stimulus. While waiting for trans-sphenoidal surgery, the treatment with ketoconazole (200mg/24h) controlled the hypercortisolism. After the surgical procedure, the patient received replacement therapy with hydrocortisone for 1 month. At the present time, the patient, without fulfilling the post surgery cure criteria, has been 2 and a half years in clinical and hormonal remission. PMID- 19282259 TI - [Evaluation of carotid intima-media thickness in familial hypercholesterolemia in childhood]. AB - INTRODUCTION: Familial hypercholesterolemia (FH) is characterized by exposure to severely elevated LDL-cholesterol from birth, which produces lipid deposits, which can be measured by means of intima-media thickness (IMT). SUBJECTS AND METHODS: The IMT and concentrations of cholesterol and its fractions, triglycerides, alipoproteins Apo-A1, Apo-B and endothelial risk factors (homocysteine and high sensitivity protein C ) were determined in 89 patients (44 males) from 2 to 19 years (9.54+/-3.91 years). IMT was measured by ultrasound using a 12MHz linear array transducer in both carotids to 1cm of the bulb. The IMT mean was compared with age, sex and analytical parameters using multiple regression analysis. RESULTS: The mean values were: IMT 0.334+/-0.088mm, total cholesterol 273.62+/-91.93mg/dl, LDL-cholesterol 204.21+/-86.16mg/dl, LDL/HDL 3.83+/-1.45, apoprotein A1 134.61+/-26.49mg/dl, apoprotein B 130.59+/-40.59mg/dl, homocysteine (median) 7.16mmol/dl, Protein C (median) 0.3mg/l. Using multiple regression analysis, only age was associated with IMT (P=0.049), a mean 0.005mm (95% CI: 0.000-0.010) being the annual increase: up to 12 years the increase in IMT was 0.002mm/year on (95% CI: -0.007-0,010) and then from that age it was 0.013mm/year (95% CI: -0.023-0.049). CONCLUSIONS: The measurement of the carotid IMT could become an objective parameter in the evaluation of the FH in childhood. In our study, it is only associated with age, the increase being most marked from 12 years onwards. PMID- 19282260 TI - Effects of filler type and content on mechanical properties of photopolymerizable composites measured across two-dimensional combinatorial arrays. AB - Multicomponent formulations coupled with complex processing conditions govern the final properties of photopolymerizable dental composites. In this study, a single test substrate was fabricated to support multiple formulations with a gradient in degree of conversion (DC), allowing the evaluation of multiple processing conditions and formulations on one specimen. Mechanical properties and damage response were evaluated as a function of filler type/content and irradiation. DC, surface roughness, modulus, hardness, scratch deformation and cytotoxicity were quantified using techniques including near-infrared spectroscopy, laser confocal scanning microscopy, depth-sensing indentation, scratch testing and cell viability. Scratch parameters (depth, width, percent recovery) were correlated to composite modulus and hardness. Total filler content, nanofiller and irradiation time/intensity all affected the final properties, with the dominant factor for improved properties being a higher DC. This combinatorial platform accelerates the screening of dental composites through the direct comparison of properties and processing conditions across the same sample. PMID- 19282261 TI - Constructing doxorubicin-loaded polymeric micelles through amphiphilic graft polyphosphazenes containing ethyl tryptophan and PEG segments. AB - By changing the molar ratio of hydrophilic and hydrophobic segments, a series of novel amphiphilic graft polyphosphazenes (PEG/EtTrp-PPPs) was synthesized via thermal ring-opening polymerization and a subsequent two-step substitution reaction of hydrophilic methoxyl polyethylene glycol (MPEG) and hydrophobic ethyl tryptophan (EtTrp). (1)H-Nuclear magnetic resonance and Fourier transform infrared studies validated the expected synthesis of copolymers. The copolymer composition was also confirmed by UV-visible spectrophotometry. The molar ratio of the segment PEG to group EtTrp was 1.33:0.67, 1.01:0.99 and 0.78:1.22, respectively. Micellization behavior of PEG/EtTrp-PPPs in an aqueous phase was characterized by fluorescence technique, dynamic light scattering and transmission electron microscopy. The critical micelle concentration (CMC) of the graft copolymer in aqueous solution was 0.158, 0.033 and 0.020gl(-1), which decreased as the hydrophobic content in amphiphilic copolymers increased. Doxorubicin (DOX) was physically loaded into micelles prepared by an O/W emulsion method with a drug loading content increasing with DOX feeding. In vitro release of DOX from micelles can be accelerated in weak acidic solution. The results of cytotoxicity study using an MTT assay method with HeLa cell showed that amphiphilic graft polyphosphazenes were biocompatible while DOX-loaded micelles achieved comparable cytotoxicity with that of free DOX. In summary, these novel amphiphilic copolymers exhibited potential to be used as injectable drug carriers for tumor treatment. PMID- 19282262 TI - Fabrication and characterization of poly(gamma-glutamic acid)-graft-chondroitin sulfate/polycaprolactone porous scaffolds for cartilage tissue engineering. AB - The development of blended biomacromolecule and polyester scaffolds can potentially be used in many tissue engineering applications. This study was to develop a poly(gamma-glutamic acid)-graft-chondroitin sulfate-blend-poly(epsilon caprolactone) (gamma-PGA-g-CS/PCL) composite biomaterial as a scaffold for cartilage tissue engineering. Chondroitin sulfate (CS) was grafted to gamma-PGA, forming a gamma-PGA-g-CS copolymer with 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide (EDC) system. The gamma-PGA-g-CS copolymers were then blended with PCL to yield a porous gamma-PGA-g-CS/PCL scaffold by salt leaching. These blended scaffolds were characterized by (1)H NMR, ESCA, water-binding capacity, mechanical test, degradation rate and CS assay. The results showed that with gamma-PGA-g-CS as a component, the water-binding capacity and the degradation rate of the scaffolds would substantially increase. During a 4 week period of culture, the mechanical stability of gamma-PGA-g-CS/PCL scaffolds was raised gradually and chondrocytes were induced to function normally in vitro. Furthermore, a larger amount of secreted GAGs was present in the gamma-PGA-g CS/PCL matrices than in the control (PCL), as revealed by Alcian blue staining of the histochemical sections. Thus, gamma-PGA-g-CS/PCL matrices exhibit excellent biodegradation and biocompatibility for chondrocytes and have potential in tissue engineering as temporary substitutes for articular cartilage regeneration. PMID- 19282263 TI - A study of properties of "micelle-enhanced" polyelectrolyte capsules: Structure, encapsulation and in vitro release. AB - "Micelle-enhanced" polyelectrolyte capsules were fabricated via a layer-by-layer technique, templated on hybrid calcium carbonate particles with built-in polymeric micelles based on polystyrene-b-poly(acrylic acid). Due to the presence of a large number of negatively charged micelles inside the polyelectrolyte capsule, which were liberated from templates, the capsule wall was reconstructed and had properties different to those of conventional polyelectrolyte capsules. This type of capsule could selectively entrap positively charged water-soluble substances. The encapsulation efficiency of positively charged substances was dependent on their molecular weight or size. For some positively charged compounds, such as rhodamine B and lysozyme, the concentration in the capsules was orders of magnitude higher than that in the incubation solution. In addition, in vitro release study suggested that the encapsulated compounds could be released through a sustained manner to a certain degree. All these results point to the fact that these capsules might be used as novel delivery systems for some water-soluble compounds. PMID- 19282264 TI - Bioactive nano-titania ceramics with biomechanical compatibility prepared by doping with piezoelectric BaTiO(3). AB - Piezoelectric BaTiO(3) was employed as a crystal growth inhibitor additive for the preparation of bioactive nano-titania ceramics in this study. It is found that the additive could significantly inhibit nano-titania ceramic crystal growth during the pressureless sintering process. This inhibitory ability has great effects on the mechanical properties and bioactivities of the nano-titania ceramics, making it possible to obtain bioactive nano-titania ceramics with mechanical properties analogous to human bone. In this study, the crystal grain sizes of the nano-titania ceramics ranged from 18 to 68nm and the particle sizes ranged from 187 to 580nm by changing the additive content from 1% to 20%. The elastic modulus of the nano-titania ceramics ranged from 6.2 to 10.6GPa, which is analogous to that of human bone, by adjusting the additive content. The piezoelectric properties of the additive also showed the enhancing effects on the bioactivity of the nano-titania ceramics, which made the osteoblasts proliferate faster on the nano-titania ceramics in cell culture experiments. It might be a potential way to prepare bioactive nano-titania ceramics with biomechanical compatibility by using BaTiO(3) as a crystal growth inhibitor. PMID- 19282265 TI - Long-term fate of transplanted autologous fat in the face. PMID- 19282266 TI - Encountering the culture of midwifery practice on the postnatal ward during action research: An impediment to change. AB - BACKGROUND: The reduction of time available to midwives during the hospital postnatal stay suggests that there is a need to review postnatal care. Innovative strategies are required which give attention to specific family needs and assist in the transition to parenthood. Nevertheless, new ideas and changes are not always readily accepted in midwifery practice. AIM: To enhance the provision of postnatal care to parents in the early postnatal period given time constraints for parents' hospital stay. METHODS: Action research was employed to explore the educational experiences of parents in the postnatal period. An action research group (ARG) was established, comprising predominantly of midwives. Based on data collected from parents through questionnaires, focus groups and interviews, three actions were developed and implemented on a postnatal ward. RESULTS: The actions were evaluated by 122 parents through self-report questionnaires. Midwives working on the postnatal ward and midwives involved in the ARG provided feedback regarding the actions through separate focus group discussions. The parents who participated in the study and, the midwives involved in the ARG, were positive about the actions and perceived them to be beneficial in preparing parents for parenthood. Many of the ward midwives, however, were negative about the actions and questioned their benefit for midwifery practice. DISCUSSION: The negativity of the ward midwives regarding the innovations implemented in the study contrasted strikingly with the positive responses from both parents and the action research midwives. Two themes emerged which may explain the response of midwives to the actions, notably, a lack of ownership of the actions and the problematic nature of the current culture of the postnatal-care environment. CONCLUSION: It appears that the provision of hospital postnatal care has been influenced by an underlying culture in midwifery practice, which in turn, has impeded the change required to enhance postnatal care. PMID- 19282267 TI - Negative priming within a stroop task in children and adolescents with attention deficit hyperactivity disorder, their siblings, and independent controls. AB - OBJECTIVE: Negative priming (NP) is the slowed response to a stimulus that was previously ignored. Response times in NP task conditions were compared with the interference provided by congruent/incongruent stimuli in a Stroop condition in the same task in children diagnosed with attention-deficit hyperactivity disorder (ADHD), their unaffected siblings, and independent controls. METHOD: Speed, accuracy, and variability of responses were compared using a computerized NP Stroop test for 35 children with ADHD, 24 siblings without diagnosis, and 37 independent healthy controls aged 6 to 17 years. RESULTS: NP was evident at test onset for congruent trials in children without a diagnosis and was reduced initially in those with ADHD occurring in the absence of a significant Stroop interference effect and independently of age or symptom severity. Incongruency masked NP effects. Cases showed more intraindividual response-time variability. CONCLUSIONS: Both NP in normal children and its reduction in ADHD cases attenuated across trials reflecting the increased facilitation from previous stimulation. PMID- 19282268 TI - Differences in accessibility, affordability, and availability (AAA) of medical specialists among three age-groups of elderly people in Israel. AB - Objectives. Older people use more health services due to health problems, but various reasons impede their ability to use health services. The purpose of this study is to examine difficulties in accessing and affording specialist services and to explore the factors that explain these difficulties among elderly people. Methods. The sample included 1,255 respondents in three age-groups: 65-75, 76-89, and 90+ years who were interviewed face-to-face in their homes. Results. The findings showed that between 21% and 41% of the respondents encountered difficulties in visiting specialists. Those aged 90+ encountered more accessibility problems and fewer affordability problems compared to their younger counterparts, and those aged 76-89 encountered more availability problems compared to the other two age-groups. Enabling and need factors were the most significant factors in explaining problems in accessing and affording specialist services. Discussion. Recommendations for policy and practice are discussed. PMID- 19282269 TI - The use of mobility devices among institutionalized older adults. AB - OBJECTIVE: The purpose of this article is to examine the demographic, health, and social characteristics of mobility device users in long-term care settings. METHODS: Data were used from a recently institutionalized sample of older adults from the Canadian Study of Health and Aging. Multinomial logistic regression was used to examine the factors associated with the use of different mobility devices (cane, walker, or wheelchair). RESULTS: Over 70% used mobility aids (over 50% used a wheelchair). Mobility limitations were strongly associated with the use of mobility devices. However, among those with mobility limitations, educational resources reduced the odds of wheelchair use. CONCLUSIONS: Consistent with findings from the community setting, need factors are strongly associated with the use of mobility aids in institutions. However, socioeconomic resources may provide older adults with alternate ways to manage mobility limitations in institutional settings. PMID- 19282270 TI - Using spirituality to cope with early-stage Alzheimer's disease. AB - Alzheimer's disease (AD) robs persons living with the disease of their independence and self-esteem, which can lead to depression, anxiety, and loneliness. Understanding how people with early-stage AD cope is a critical step in enhancing their adaptive abilities and ultimately improving their quality of life. This qualitative study describes how individuals with early-stage AD use spirituality to cope with the losses of self-esteem, independence, and social interaction that they face. The purposive sample for this focused ethnographic study consisted of 15 participants living at home in central Arkansas. Holding onto faith, seeking reassurance and hope, and staying connected were the global themes. Personal faith, prayer, connection to church, and family support enhanced the ability of people with early-stage AD to keep a positive attitude as they face living with AD. PMID- 19282271 TI - The nursing practice environment and nurse-perceived quality of geriatric care in hospitals. AB - The relationships between general and geriatric-specific nursing practice environments (NPEs) and nurse-perceived quality of geriatric care in hospitals were examined using the Nurses Improving Care for Healthsystems Elders benchmarking database. The overall general NPE was negatively related, but the overall geriatric-specific NPE was positively related to quality of geriatric care. Among five subdomains of the general NPE measured by the Practice Environment Scale of the Nursing Work Index, Nurse Participation in Hospital Affairs was positively related to quality of geriatric care, whereas two subdomains were not significant, and another two were negatively related to quality of geriatric care. All three subdomains of the geriatric-specific NPE measured by the Geriatric Nursing Practice Environment scale were positively related to quality of geriatric care when adjusting for general NPE. These findings suggest geriatric-specific organizational support combined with nurse involvement in hospital decision making is critical for delivering quality geriatric care. PMID- 19282272 TI - Mechanism of inhibition defines CETP activity: a mathematical model for CETP in vitro. AB - Because cholesteryl ester transfer protein (CETP) inhibition is a potential HDL raising therapy, interest has been raised in the mechanisms and consequences of CETP activity. To explore these mechanisms and the dynamics of CETP in vitro, a mechanistic mathematical model was developed based upon the shuttle mechanism for lipid transfer. Model parameters were estimated from eight published experimental datasets, and the resulting model captures observed dynamics of CETP in vitro. Simulations suggest the shuttle mechanism yields behaviors consistent with experimental observations. Three key findings predicted from model simulations are: 1) net CE transfer activity from HDL to VLDL and LDL can be significantly altered by changing the balance of homoexchange versus heteroexchange of neutral lipids via CETP; 2) lipemia-induced increases in CETP activity are more likely caused by increases in lipoprotein particle size than particle number; and 3) the inhibition mechanisms of the CETP inhibitors torcetrapib and JTT-705 are significantly more potent than a classic competitive inhibition mechanism with the irreversible binding mechanism having the most robust response. In summary, the model provides a plausible representation of CETP activity in vitro, corroborates strong evidence for the shuttle hypothesis, and provides new insights into the consequences of CETP activity and inhibition on lipoproteins. PMID- 19282273 TI - Differential activity-based gel electrophoresis for comparative analysis of lipolytic and esterolytic activities. AB - We established a novel technique for differential activity-based gel electrophoresis (DABGE) of lipolytic enzymes from two different biological samples. For this purpose, a set of three fluorescent suicide inhibitors was developed. These probes possess the same substrate analogous structures but carry different cyanine dyes (Cy2b, Cy3, and Cy5) as reporter fluorophores. For comparison of enzyme profiles, two samples are individually labeled with a different probe followed by mixing, gel electrophoresis, fluorescence imaging, and identification of the tagged proteins by MS/MS. Protocols for quantitative determination of active enzymes were developed on the basis of lipolytic proteomes that had been admixed with defined amounts of known lipases and esterases. A detailed analysis of the fluorescence intensities showed that the found enzyme ratios very closely reflected the relative amounts of the labeled enzymes that were used for spiking. The DABGE method was used to compare the lipolytic proteomes of brown and white adipose tissue showing specific enzyme patterns of both samples. This study represents the first application of this technology for comparative analysis of lipases and esterases. Further applications of this technique can be expected to provide entirely new information on lipid enzymology in health and disease with high precision. PMID- 19282274 TI - Risks of cardiovascular events and effects of routine blood pressure lowering among patients with type 2 diabetes and atrial fibrillation: results of the ADVANCE study. AB - AIMS: The aim of this study was to investigate serious clinical outcomes associated with atrial fibrillation (AF) and the effects of routine blood pressure lowering on such outcomes in the presence or absence of AF, among individuals with type 2 diabetes. METHODS AND RESULTS: About 11 140 patients with type 2 diabetes (7.6% of whom had AF at baseline) were randomized to a fixed combination of perindopril and indapamide or placebo in the Action in Diabetes and Vascular Disease: preterAx and diamicroN-MR Controlled Evaluation (ADVANCE) study. We compared total mortality and cardiovascular disease outcomes and effects of randomized treatment for 4.3 years on such outcomes between patients with and without AF at baseline. After multiple adjustments, AF was associated with a 61% (95% confidence interval 31-96, P < 0.0001) greater risk of all-cause mortality and comparable higher risks of cardiovascular death, stroke, and heart failure (all P < 0.001). Routine treatment with a fixed combination of perindopril and indapamide produced similar relative, but greater absolute, risk reductions for all-cause and cardiovascular mortalities in patients with AF, compared with those without AF. The number of patients needed to be treated with perindopril-indapamide for 5 years to prevent one cardiovascular death was 42 for patients with AF and 120 for patients without AF at baseline. CONCLUSION: Atrial fibrillation is relatively common in type 2 diabetes and is associated with substantially increased risks of death and cardiovascular events in patients with type 2 diabetes. This arrhythmia identifies individuals who are likely to obtain greater absolute benefits from blood pressure-lowering treatment. Atrial fibrillation in diabetic patients should be regarded as a marker of particularly adverse outcome and prompt aggressive management of all risk factors. PMID- 19282275 TI - Elevated plasma stromal cell-derived factor 1 protein and its gene polymorphism in patients with pelvic inflammatory disease. AB - The objective was to compare the expression of plasma stromal cell-derived factor 1 and the gene polymorphism in patients with pelvic inflammatory disease and healthy controls. The enzyme-linked immunosorbent assay and polymerase chain reaction-restriction fragment length polymorphism were, respectively, used to measure the plasma stromal cell-derived factor 1alpha level and stromal cell derived factor 1 polymorphism in 50 healthy controls and in 44 patients with pelvic inflammatory disease before and after they received routine treatment protocols. The level of plasma stromal cell-derived factor 1alpha was elevated in patients with pelvic inflammatory disease compared to normal controls and decreased significantly after treatment. There were significant correlations between plasma stromal cell-derived factor 1alpha level and neutrophil count as well as between stromal cell-derived factor 1alpha level and white blood cell count in patients with pelvic inflammatory disease. There was no significantly different distribution of stromal cell-derived factor 1 genotypes between patients with pelvic inflammatory disease and normal controls. Patients with pelvic inflammatory disease having stromal cell-derived factor 1-3'A allele were associated with significantly elevated plasma stromal cell-derived factor 1alpha concentration compared to patients with pelvic inflammatory disease having G/G homozygous alleles (P < .02). In normal controls, there was no significant difference in the plasma stromal cell-derived factor 1 level between individuals with and without stromal cell-derived factor 1-3'A allele. When the cutoff level of plasma stromal cell-derived factor 1alpha level was determined to be 2192 pg/mL based on receiver-operating characteristic curve, the sensitivity, specificity, positive predictive value, and negative predictive value as well as accuracy were 77.3%, 88.0%, 85.0%, 81.5%, and 83.0%. In conclusion, when the cutoff level was determined to be 2192 pg/mL, plasma stromal cell-derived factor 1alpha level can be used to predict pelvic inflammatory disease. PMID- 19282276 TI - Picture quiz. Imaging after trauma to the pelvis and hip. PMID- 19282277 TI - Picture quiz. Something's in the air. PMID- 19282279 TI - Biochemical characterization and membrane topology of Alg2 from Saccharomyces cerevisiae as a bifunctional alpha1,3- and 1,6-mannosyltransferase involved in lipid-linked oligosaccharide biosynthesis. AB - N-Linked glycosylation involves the ordered, stepwise synthesis of the unique lipid-linked oligosaccharide precursor Glc(3)Man(9) GlcNAc(2)-PP-Dol on the endoplasmic reticulum (ER), catalyzed by a series of glycosyltransferases. Here we characterize Alg2 as a bifunctional enzyme that is required for both the transfer of the alpha1,3- and the alpha1,6-mannose-linked residue from GDP mannose to Man(1)GlcNAc(2)-PP-Dol forming the Man(3)GlcNAc(2)-PP-Dol intermediate on the cytosolic side of the ER. Alg2 has a calculated mass of 58 kDa and is predicted to contain four transmembrane-spanning helices, two at the N terminus and two at the C terminus. Contradictory to topology predictions, we prove that only the two N-terminal domains fulfill this criterion, whereas the C-terminal hydrophobic sequences contribute to ER localization in a nontransmembrane manner. Surprisingly, none of the four domains is essential for transferase activity because truncated Alg2 variants can exert their function as long as Alg2 is associated with the ER by either its N- or C-terminal hydrophobic regions. By site-directed mutagenesis we demonstrate that an EX(7)E motif, conserved in a variety of glycosyltransferases, is not important for Alg2 function in vivo and in vitro. Instead, we identify a conserved lysine residue, Lys(230), as being essential for activity, which could be involved in the binding of the phosphate of the glycosyl donor. PMID- 19282280 TI - Neurosteroids allosterically modulate binding of the anesthetic etomidate to gamma-aminobutyric acid type A receptors. AB - Photoaffinity labeling of gamma-aminobutyric acid type A (GABA(A))-receptors (GABA(A)R) with an etomidate analog and mutational analyses of direct activation of GABA(A)R by neurosteroids have each led to the proposal that these structurally distinct general anesthetics bind to sites in GABA(A)Rs in the transmembrane domain at the interface between the beta and alpha subunits. We tested whether the two ligand binding sites might overlap by examining whether neuroactive steroids inhibited etomidate analog photolabeling. We previously identified (Li, G. D., Chiara, D. C., Sawyer, G. W., Husain, S. S., Olsen, R. W., and Cohen, J. B. (2006) J. Neurosci. 26, 11599-11605) azietomidate photolabeling of GABA(A)R alpha1Met-236 and betaMet-286 (in alphaM1 and betaM3). Positioning these two photolabeled amino acids in a single type of binding site at the interface of beta and alpha subunits (two copies per pentamer) is consistent with a GABA(A)R homology model based upon the structure of the nicotinic acetylcholine receptor and with recent alphaM1 to betaM3 cross-linking data. Biologically active neurosteroids enhance rather than inhibit azietomidate photolabeling, as assayed at the level of GABA(A)R subunits on analytical SDS-PAGE, and protein microsequencing establishes that the GABA(A)R-modulating neurosteroids do not inhibit photolabeling of GABA(A)R alpha1Met-236 or betaMet-286 but enhance labeling of alpha1Met-236. Thus modulatory steroids do not bind at the same site as etomidate, and neither of the amino acids identified as neurosteroid activation determinants (Hosie, A. M., Wilkins, M. E., da Silva, H. M., and Smart, T. G. (2006) Nature 444, 486-489) are located at the subunit interface defined by our etomidate site model. PMID- 19282281 TI - Molecular basis for class V beta-tubulin effects on microtubule assembly and paclitaxel resistance. AB - Vertebrates produce at least seven distinct beta-tubulin isotypes that coassemble into all cellular microtubules. The functional differences among these tubulin isoforms are largely unknown, but recent studies indicate that tubulin composition can affect microtubule properties and cellular microtubule-dependent behavior. One of the isotypes whose incorporation causes the largest change in microtubule assembly is beta5-tubulin. Overexpression of this isotype can almost completely destroy the microtubule network, yet it appears to be required in smaller amounts for normal mitotic progression. Moderate levels of overexpression can also confer paclitaxel resistance. Experiments using chimeric constructs and site-directed mutagenesis now indicate that the hypervariable C-terminal region of beta5 plays no role in these phenotypes. Instead, we demonstrate that two residues found in beta5 (Ser-239 and Ser-365) are each sufficient to inhibit microtubule assembly and confer paclitaxel resistance when introduced into beta1 tubulin; yet the single mutation of residue Ser-239 in beta5 eliminates its ability to confer these phenotypes. Despite the high degree of conservation among beta-tubulin isotypes, mutations affecting residue 365 demonstrate that amino acid substitutions can be context sensitive; i.e. an amino acid change in one isotype will not necessarily produce the same phenotype when introduced into a different isotype. Modeling studies indicate that residue Cys-239 of beta1 tubulin is close to a highly conserved Cys-354 residue suggesting the possibility that disulfide formation could play a significant role in the stability of microtubules formed with beta1- but not with beta5-tubulin. PMID- 19282282 TI - Single molecule force spectroscopy of the cardiac titin N2B element: effects of the molecular chaperone alphaB-crystallin with disease-causing mutations. AB - The small heat shock protein alphaB-crystallin interacts with N2B-Us, a large unique sequence found in the N2B element of cardiac titin. Using single molecule force spectroscopy, we studied the effect of alphaB-crystallin on the N2B-Us and its flanking Ig-like domains. Ig domains from the proximal tandem Ig segment of titin were also studied. The effect of wild type alphaB-crystallin on the single molecule force-extension curve was determined as well as that of mutant alphaB crystallins harboring the dilated cardiomyopathy missense mutation, R157H, or the desmin-related myopathy mutation, R120G. Results revealed that wild type alphaB crystallin decreased the persistence length of the N2B-Us (from approximately 0.7 to approximately 0.2 nm) but did not alter its contour length. alphaB-crystallin also increased the unfolding force of the Ig domains that flank the N2B-Us (by 51 +/- 3 piconewtons); the rate constant of unfolding at zero force was estimated to be approximately 17-fold lower in the presence of alphaB-crystallin (1.4 x 10(-4) s(-1) versus 2.4 x 10(-3) s(-1)). We also found that alphaB-crystallin increased the unfolding force of Ig domains from the proximal tandem Ig segment by 28 +/- 6 piconewtons. The effects of alphaB-crystallin were attenuated by the R157H mutation (but were still significant) and were absent when using the R120G mutant. We conclude that alphaB-crystallin protects titin from damage by lowering the persistence length of the N2B-Us and reducing the Ig domain unfolding probability. Our finding that this effect is either attenuated (R157H) or lost (R120G) in disease causing alphaB-crystallin mutations suggests that the interaction between alphaB-crystallin and titin is important for normal heart function. PMID- 19282283 TI - Interdomain flexibility in full-length matrix metalloproteinase-1 (MMP-1). AB - The presence of extensive reciprocal conformational freedom between the catalytic and the hemopexin-like domains of full-length matrix metalloproteinase-1 (MMP-1) is demonstrated by NMR and small angle x-ray scattering experiments. This finding is discussed in relation to the essentiality of the hemopexin-like domain for the collagenolytic activity of MMP-1. The conformational freedom experienced by the present system, having the shortest linker between the two domains, when compared with similar findings on MMP-12 and MMP-9 having longer and the longest linker within the family, respectively, suggests this type of conformational freedom to be a general property of all MMPs. PMID- 19282284 TI - Characterization of WbpB, WbpE, and WbpD and reconstitution of a pathway for the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid in Pseudomonas aeruginosa. AB - The lipopolysaccharide of Pseudomonas aeruginosa PAO1 contains an unusual sugar, 2,3-diacetamido-2,3-dideoxy-d-mannuronic acid (d-ManNAc3NAcA). wbpB, wbpE, and wbpD are thought to encode oxidase, transaminase, and N-acetyltransferase enzymes. To characterize their functions, recombinant proteins were overexpressed and purified from heterologous hosts. Activities of His(6)-WbpB and His(6)-WbpE were detected only when both proteins were combined in the same reaction. Using a direct MALDI-TOF mass spectrometry approach, we identified ions that corresponded to the predicted products of WbpB (UDP-3-keto-d-GlcNAcA) and WbpE (UDP-d GlcNAc3NA) in the coupled enzyme-substrate reaction. Additionally, in reactions involving WbpB, WbpE, and WbpD, an ion consistent with the expected product of WbpD (UDP-d-GlcNAc3NAcA) was identified. Preparative quantities of UDP-d GlcNAc3NA and UDP-d-GlcNAc3NAcA were enzymatically synthesized. These compounds were purified by high-performance liquid chromatography, and their structures were elucidated by NMR spectroscopy. This is the first report of the functional characterization of these proteins, and the enzymatic synthesis of UDP-d GlcNAc3NA and UDP-d-GlcNAc3NAcA. PMID- 19282285 TI - Analysis of intracellular substrates and products of thimet oligopeptidase in human embryonic kidney 293 cells. AB - Thimet oligopeptidase (EC 3.4.24.15; EP24.15) is an intracellular enzyme that has been proposed to metabolize peptides within cells, thereby affecting antigen presentation and G protein-coupled receptor signal transduction. However, only a small number of intracellular substrates of EP24.15 have been reported previously. Here we have identified over 100 peptides in human embryonic kidney 293 (HEK293) cells that are derived from intracellular proteins; many but not all of these peptides are substrates or products of EP24.15. First, cellular peptides were extracted from HEK293 cells and incubated in vitro with purified EP24.15. Then the peptides were labeled with isotopic tags and analyzed by mass spectrometry to obtain quantitative data on the extent of cleavage. A related series of experiments tested the effect of overexpression of EP24.15 on the cellular levels of peptides in HEK293 cells. Finally, synthetic peptides that corresponded to 10 of the cellular peptides were incubated with purified EP24.15 in vitro, and the cleavage was monitored by high pressure liquid chromatography and mass spectrometry. Many of the EP24.15 substrates identified by these approaches are 9-11 amino acids in length, supporting the proposal that EP24.15 can function in the degradation of peptides that could be used for antigen presentation. However, EP24.15 also converts some peptides into products that are 8-10 amino acids, thus contributing to the formation of peptides for antigen presentation. In addition, the intracellular peptides described here are potential candidates to regulate protein interactions within cells. PMID- 19282286 TI - Nova-1 mediates glucocorticoid-induced inhibition of pre-mRNA splicing of gonadotropin-releasing hormone transcripts. AB - Glucocorticoid (GC) is known to affect the reproductive system by suppressing the gonadotropin-releasing hormone (GnRH) gene expression in the hypothalamus. However, the mechanism of this effect is poorly understood. We show here that the GC-induced reduction of GnRH mRNA is due to attenuation of a post-transcriptional process i.e. splicing of intron A. Treatment of dexamethasone (DEX), a synthetic GC, lowered GnRH mRNA transcripts and was accompanied by reduced excision of the first intron (intron A) from the GnRH pre-mRNA both in vitro and in vivo. While seeking to identify the splicing factors involved in GC-inhibited GnRH pre-mRNA splicing, we found that DEX down-regulated neuro-oncological ventral antigen-1 (Nova-1) mRNA and protein and that knockdown of Nova-1 reduced intron A excision from GnRH pre-mRNA. Nova-1 overexpression reversed the DEX-induced reduction of intron A excision. Nova-1 appears to promote intron A excision by binding to the distal region of exon 1 of the GnRH pre-mRNA. Taken together, our findings indicate that the intron A excision by Nova-1 is a target of GC for down regulation of GnRH gene expression, and more importantly, we characterized Nova 1, a brain-enriched splicing regulator responsible for GnRH pre-mRNA splicing. PMID- 19282288 TI - Propagation of tau misfolding from the outside to the inside of a cell. AB - Tauopathies are neurodegenerative diseases characterized by aggregation of the microtubule-associated protein Tau in neurons and glia. Although Tau is normally considered an intracellular protein, Tau aggregates are observed in the extracellular space, and Tau peptide is readily detected in the cerebrospinal fluid of patients. Tau aggregation occurs in many diseases, including Alzheimer disease and frontotemporal dementia. Tau pathology begins in discrete, disease specific regions but eventually involves much larger areas of the brain. It is unknown how this propagation of Tau misfolding occurs. We hypothesize that extracellular Tau aggregates can transmit a misfolded state from the outside to the inside of a cell, similar to prions. Here we show that extracellular Tau aggregates, but not monomer, are taken up by cultured cells. Internalized Tau aggregates displace tubulin, co-localize with dextran, a marker of fluid-phase endocytosis, and induce fibrillization of intracellular full-length Tau. These intracellular fibrils are competent to seed fibril formation of recombinant Tau monomer in vitro. Finally, we observed that newly aggregated intracellular Tau transfers between co-cultured cells. Our data indicate that Tau aggregates can propagate a fibrillar, misfolded state from the outside to the inside of a cell. This may have important implications for understanding how protein misfolding spreads through the brains of tauopathy patients, and it is potentially relevant to myriad neurodegenerative diseases associated with protein misfolding. PMID- 19282287 TI - Ikaros stability and pericentromeric localization are regulated by protein phosphatase 1. AB - Ikaros encodes a zinc finger protein that is involved in gene regulation and chromatin remodeling. The majority of Ikaros localizes at pericentromeric heterochromatin (PC-HC) where it regulates expression of target genes. Ikaros function is controlled by posttranslational modification. Phosphorylation of Ikaros by CK2 kinase determines its ability to bind DNA and exert cell cycle control as well as its subcellular localization. We report that Ikaros interacts with protein phosphatase 1 (PP1) via a conserved PP1 binding motif, RVXF, in the C-terminal end of the Ikaros protein. Point mutations of the RVXF motif abolish Ikaros-PP1 interaction and result in decreased DNA binding, an inability to localize to PC-HC, and rapid degradation of the Ikaros protein. The introduction of alanine mutations at CK2-phosphorylated residues increases the half-life of the PP1-nonbinding Ikaros mutant. This suggests that dephosphorylation of these sites by PP1 stabilizes the Ikaros protein and prevents its degradation. In the nucleus, Ikaros forms complexes with ubiquitin, providing evidence that Ikaros degradation involves the ubiquitin/proteasome pathway. In vivo, Ikaros can target PP1 to the nucleus, and a fraction of PP1 colocalizes with Ikaros at PC-HC. These data suggest a novel function for the Ikaros protein; that is, the targeting of PP1 to PC-HC and other chromatin structures. We propose a model whereby the function of Ikaros is controlled by the CK2 and PP1 pathways and that a balance between these two signal transduction pathways is essential for normal cellular function and for the prevention of malignant transformation. PMID- 19282289 TI - Type II cGMP-dependent protein kinase mediates osteoblast mechanotransduction. AB - Continuous bone remodeling in response to mechanical loading is critical for skeletal integrity, and interstitial fluid flow is an important stimulus for osteoblast/osteocyte growth and differentiation. However, the biochemical signals mediating osteoblast anabolic responses to mechanical stimulation are incompletely understood. In primary human osteoblasts and murine MC3T3-E1 cells, we found that fluid shear stress induced rapid expression of c-fos, fra-1, fra-2, and fosB/DeltafosB mRNAs; these genes encode transcriptional regulators that maintain skeletal integrity. Fluid shear stress increased osteoblast nitric oxide (NO) synthesis, leading to activation of cGMP-dependent protein kinase (PKG). Pharmacological inhibition of the NO/cGMP/PKG signaling pathway blocked shear induced expression of all four fos family genes. Induction of these genes required signaling through MEK/Erk, and Erk activation was NO/cGMP/PKG-dependent. Treating cells with a membrane-permeable cGMP analog partly mimicked the effects of fluid shear stress on Erk activity and fos family gene expression. In cells transfected with small interfering RNAs (siRNA) specific for membrane-bound PKG II, shear- and cGMP-induced Erk activation and fos family gene expression was nearly abolished and could be restored by transducing cells with a virus encoding an siRNA-resistant form of PKG II; in contrast, siRNA-mediated repression of the more abundant cytosolic PKG I isoform was without effect. Thus, we report a novel function for PKG II in osteoblast mechanotransduction, and we propose a model whereby NO/cGMP/PKG II-mediated Erk activation and induction of c-fos, fra-1, fra 2, and fosB/DeltafosB play a key role in the osteoblast anabolic response to mechanical stimulation. PMID- 19282290 TI - Heterogeneous nuclear ribonucleoprotein G regulates splice site selection by binding to CC(A/C)-rich regions in pre-mRNA. AB - Almost every protein-coding gene undergoes pre-mRNA splicing, and the majority of these pre-mRNAs are alternatively spliced. Alternative exon usage is regulated by the transient formation of protein complexes on the pre-mRNA that typically contain heterogeneous nuclear ribonucleoproteins (hnRNPs). Here we characterize hnRNP G, a member of the hnRNP class of proteins. We show that hnRNP G is a nuclear protein that is expressed in different concentrations in various tissues and that interacts with other splicing regulatory proteins. hnRNP G is part of the supraspliceosome, where it regulates alternative splice site selection in a concentration-dependent manner. Its action on alternative exons can occur without a functional RNA-recognition motif by binding to other splicing regulatory proteins. The RNA-recognition motif of hnRNP G binds to a loose consensus sequence containing a CC(A/C) motif, and hnRNP G preferentially regulates alternative exons where this motif is clustered in close proximity. The X chromosomally encoded hnRNP G regulates different RNAs than its Y-chromosomal paralogue RNA-binding motif protein, Y-linked (RBMY), suggesting that differences in alternative splicing, evoked by the sex-specific expression of hnRNP G and RBMY, could contribute to molecular sex differences in mammals. PMID- 19282291 TI - Mature solid teratoma of the fallopian tube mimicking metastasis of endometrial adenocarcinoma: a case report. AB - We report a mature solid teratoma of the fallopian tube in an elderly woman. This mass was noted on CT scan and considered metastatic in nature since following a bioptical diagnosis of endometrial adenocarcinoma. Hysterectomy and bilateral salpingectomy and ovariectomy were performed and a second minor mature solid teratoma was discovered inside the right ovary. Neoplasms of the fallopian tube are very uncommon and this is the first Italian case to be added to about other 60 of the literature. Moreover it joins minimum teratomatous diameter with maximum recorded age. PMID- 19282292 TI - Breast carcinoma with chondroid differentiation: a clinicopathologic study of 21 triple negative (ER-, PR-, Her2/neu-) cases. AB - A total of 21 metaplastic breast carcinomas (MCs) with chondroid differentiation were evaluated to better establish the clinicopathological features of this variant. The tumors exhibited mainly invasive carcinoma admixed with areas of cartilaginous matrix production. An associated ductal intraepithelial neoplasia component grade 2 or 3 was observed in 43% of cases. Immunohistochemical analysis revealed a triple negative (ER-, PR-, and Her2/neu-) immunoprofile and no expression of the androgen receptor. EGFR-positivity was found in 88% of evaluated cases, consistent with the proposed basaloid phenotype for all MC. Compared with previous studies that reviewed MC with osseus and cartilaginous elements, the incidence of axillary lymph node metastasis was significantly higher in our study and 60% of positive nodes exhibited chondroid differentiation. Available follow-up data (n = 10) revealed aggressive behavior of this MC variant with frequent metastasis, including visceral involvement and local chest wall recurrence despite chemotherapy and radiation. Three patients subsequently died of metastatic disease. PMID- 19282293 TI - Endosalpingiosis of the urinary bladder: a case of probable implantative origin with characterization of benign Fallopian tube immunohistochemistry. AB - Mullerianosis of the bladder is an infrequently described lesion consisting of multiple Mullerian-type tissues within the urinary bladder. Few previous cases of pure endosalpingiosis have been described. Here we present a 54-year-old post menopausal female with a history of prior pelvic surgery with traumatic bladder injury, who was found to have a cystic lesion in the posterior wall of the bladder. Routine histology demonstrated cyst epithelium characteristic of endosalpingiosis. Three benign Fallopian tube specimens were obtained and stained with the relevant immunohistochemical markers for comparison. Results showed an identical immunohistochemical profile between the bladder cyst lining and the normal Fallopian tube controls. This case represents a rare instance of pure endosalpingiosis of the urinary bladder, with a likely implantative origin. This form of bladder Mullerianosis should therefore be considered within the differential diagnosis of cystic lesions of the bladder. PMID- 19282294 TI - Primary pulmonary osteosarcoma: a report of 4 cases and a review of the literature. AB - Primary pulmonary osteosarcoma is very rare. Most cases are secondary deposits from primaries arising in the appendicular skeleton. Four cases of primary osteogenic sarcoma of the lung are described and the literature reviewed for previously reported cases. These pulmonary tumors occur in patients who are in their fourth to seventh decades, that is, an older age group than their primary bone equivalent. There is a slight male predominance. There appears to be a propensity for the left lung, especially the left upper lobe. The clinical presentation is similar to primary (epithelial) lung cancer. Differentiation from pleomorphic carcinomas and other sarcomas is discussed. We know of no predisposing factor(s) in our cases for the development of this tumor. PMID- 19282295 TI - Hepatic angiomyolipoma and intramural small intestinal schwannoma: a coincidence or a relationship? AB - We report a case of hepatic angiomyolipoma associated to a small bowel schwannoma in a 40-years old woman. Both lesions were asyntomatic. Histologically, hepatic angiomyolipoma showed oncocytic features and scanty adipose tissue, the tumor cells expressed desmin, smooth muscle actin, S-100 protein and HMB45. The tumor cells of intramural small intestinal mass were positive for S-100 protein and GFAP and negative for CD117, CD34 and desmin. To the best of our knowledge, no case of hepatic angiomyolipoma has been previously reported in association with intestinal schwannoma. PMID- 19282296 TI - The pathogenetic and prognostic value of biologic markers in acute lung injury. AB - Over the past 2 decades, measurement of biomarkers in both the airspaces and plasma early in the course of acute lung injury has provided new insights into the mechanisms of lung injury. In addition, biologic markers of cell-specific injury, acute inflammation, and altered coagulation correlate with mortality from acute lung injury in several single center studies as well as in multicenter clinical trials. To date, biomarkers have been measured largely for research purposes. However, with improved understanding of their role in the pathogenesis of acute lung injury, biomarkers may play an important role in early detection of lung injury, risk stratification for clinical trials, and, ultimately, tailoring specific therapies to individual patients. This article provides a review of biologic markers in acute lung injury, with an emphasis on recent analysis of results from multicenter clinical trials. PMID- 19282297 TI - Decannulation following tracheostomy for prolonged mechanical ventilation. AB - BACKGROUND: We examined the process of decannulation following tracheostomy in patients transferred to a long-term acute care (LTAC) hospital for weaning from prolonged mechanical ventilation (PMV). METHODS: A retrospective chart review of 135 patients. RESULTS: Decannulation was successful in 35% of patients a median of 45 days (IQR, 32-76) following tracheostomy. Patients who failed decannulation had a tracheostomy tube placed earlier (14 days; IQR 11-18 vs. 18 days; IQR 14 30, P=.04) and had a shorter length of stay at the acute facility (20 days; IQR, 16-23 vs. 31 days; IQR 24-45, P=.003) compared with patients who were decannulated. Length of stay and cost of care at the LTAC did not differ with decannulation status. At 3.5 years, 35% (47/135) of all patients and 62% (29/47) of decannulated patients were alive. CONCLUSIONS: Decannulation was achieved in 35% of patients transferred to an LTAC for weaning from prolonged mechanical ventilation. PMID- 19282298 TI - Malignant Osborn waves during therapeutic hypothermia. PMID- 19282299 TI - Rocuronium dose-dependently suppresses the spectral entropy response to tracheal intubation during propofol anaesthesia. AB - BACKGROUND: The Entropy Module anaesthesia monitor displays two spectral entropy based indices, response entropy (RE) and state entropy (SE). The difference between RE and SE (RE-SE), which mainly reflects electromyography activation, is thought to indicate the adequacy of antinociception. Little is known, however, about the effects of neuromuscular blocking agents on the RE-SE. We investigated the effects of rocuronium on the RE-SE response to tracheal intubation. METHODS: Forty-four patients were randomly assigned to receive one of four rocuronium doses (0.3, 0.6, 0.9, and 1.2 mg kg(-1)). Anaesthesia was induced by propofol target-controlled infusion. Rocuronium was administered 2 min after anaesthesia induction. Tracheal intubation was performed 7 min after anaesthesia induction. Arterial pressure, heart rate (HR), bispectral index (BIS), RE, SE, and patient movement were recorded. RESULTS: All EEG-derived indices (BIS, RE, SE, and RE-SE) increased after tracheal intubation. The maximum increase in the indices after tracheal intubation was significantly suppressed by an increase in the rocuronium dose. Patient movement after tracheal intubation was suppressed by an increase in the rocuronium dose. All indices were higher in patients who moved during or after tracheal intubation than in those who did not move. Rocuronium dose did not affect the mean arterial pressure or HR in response to tracheal intubation. CONCLUSIONS: The RE-SE response to tracheal intubation was suppressed by increasing the rocuronium dose. Estimates of nociception using RE-SE should be interpreted carefully in different states of muscle paralysis during general anaesthesia. PMID- 19282300 TI - Efficacy of N-acetylcysteine in preventing renal injury after heart surgery: a systematic review of randomized trials. AB - AIMS: The aim of this study was to assess whether perioperative N-acetylcysteine (NAC), an antioxidant, prevents acute renal injury (ARI) after cardiac surgery. METHODS AND RESULTS: We performed a systematic review of randomized controlled trials (RCTs) of NAC in adult cardiac surgery patients. The RCTs were identified by searching MEDLINE (1960-2008), clinicaltrials.gov website, and hand-searching references of relevant publications. Primary outcome was ARI (absolute increase >0.5 mg/dL or relative increase >25%, in serum creatinine from baseline within 5 days after surgery). Random effects model was used to perform a meta-analysis. Forest plots and I(2) test were used to assess heterogeneity among studies. Ten RCTs (n = 1163 patients) were included. Mean age was 70 +/- 7.4 years, 71% were male, and 66% underwent coronary artery bypass surgery. N-Acetylcysteine did not reduce ARI incidence [35% NAC vs. 37% placebo; relative risk (RR) 0.91, 95% CI 0.79-1.06, P = 0.24]. Overall, 3.3% of patients required haemodialysis (NAC vs. placebo; RR = 1.13, 95% CI 0.59-2.17) and 3% died (RR = 1.10, 95% CI 0.56-2.16). There was a trend towards reduced ARI incidence among patients with baseline chronic kidney disease assigned to intravenous NAC (RR = 0.80, 95% CI 0.64-1.01, P = 0.06). CONCLUSION: This meta-analysis of RCTs showed that prophylactic perioperative NAC in cardiac surgery does not reduce ARI, haemodialysis, or death. PMID- 19282301 TI - Blindness and laser photocoagulation in patients with childhood-onset type 1 diabetes in Japan. AB - AIM: The aim of the study was to investigate trends in the incidence of blindness and the association with laser photocoagulation in patients with type 1 diabetes in Japan. METHODS: Patients diagnosed between 1965 and 1979 aged under 18 years old were studied. The status of blindness and laser photocoagulation was identified as of 1 January 1995. To examine the time trend, we divided the cohort into two groups: 285 patients diagnosed between 1965 and 1969 (65-69 cohort) and 769 patients diagnosed between 1975 and 1979 (75-79 cohort). Survival analysis was performed using the Kaplan-Meier method. Cox proportional hazard models were used to assess the demographic characteristics. RESULTS: Blindness developed in 60 subjects in the 65-69 cohort and 15 subjects in the 75-79 cohort. The incidence of blindness in the 75-79 cohort was significantly lower than that in the 65-69 cohort (p<0.0001). In spite of no change in the use of laser photocoagulation in the 75-79 cohort compared with the 65-69 cohort, the hazard ratio for the blindness in those who received laser photocoagulation in the 75-79 cohort decreased significantly to 0.55 (p<0.01) compared with those in the 65-69 cohort when adjusted for the age of onset, sex, and time of diagnosis. CONCLUSION: The incidence of blindness decreased significantly for the subjects diagnosed more recently. The change in quality and the earlier introduction of laser photocoagulation might have contributed to the decreased incidence of blindness observed over time. PMID- 19282302 TI - Steroid profiles of professional soccer players: an international comparative study. AB - BACKGROUND AND OBJECTIVES: Urinary steroid profiling is used in doping controls to detect testosterone abuse. A testosterone over epitestosterone (T/E) ratio exceeding 4.0 is considered as suspicious of testosterone administration, irrespectively of individual heterogeneous factors such as the athlete's ethnicity. A deletion polymorphism in the UGT2B17 gene was demonstrated to account for a significant part of the interindividual variability in the T/E between Caucasians and Asians. Here, the variability of urinary steroid profiles was examined in a widely heterogeneous cohort of professional soccer players. METHOD: The steroid profile of 57 Africans, 32 Asians, 50 Caucasians and 32 Hispanics was determined by gas chromatography-mass spectrometry. RESULTS: Significant differences have been observed between all ethnic groups. After estimation of the prevalence of the UGT2B17 deletion/deletion genotype (African: 22%; Asian: 81%; Caucasian: 10%; Hispanic: 7%), ethnic-specific thresholds were developed for a specificity of 99% for the T/E (African: 5.6; Asian: 3.8; Caucasian: 5.7; Hispanic: 5.8). Finally, another polymorphism could be hypothesised in Asians based on specific concentration ratio of 5alpha-/5beta androstane-3alpha,17beta-diol in urine. CONCLUSION: These results demonstrate that a unique and non-specific threshold to evidence testosterone misuse is not fit for purpose. An athlete's endocrinological passport consisting of a longitudinal follow-up together with the ethnicity and/or the genotype would strongly enhance the detection of testosterone abuse. Finally, additional genotyping studies should be undertaken to determine whether the remaining unexplained disparities have an environmental or a genetic origin. PMID- 19282303 TI - Hip flexibility and strength measures: reliability and association with athletic groin pain. AB - OBJECTIVE: Groin pain commonly affects football players and can be associated with prolonged recovery periods. Understanding the relationship between groin pain and reliable measures of hip flexibility and strength may facilitate the development of optimal rehabilitation and prevention strategies. In this study, the reliability and association with athletic groin pain of hip flexibility and strength measures were investigated. METHODS: A cohort of 29 football players (15 21 years) participating in junior elite competitions (Australian Rules football and soccer) were recruited. The intra-rater reliability (n=13) and inter-rater reliability (n=12) of various hip flexibility (bent knee fall out test, hip internal rotation, hip external rotation) and strength (hip abduction, hip internal rotation, hip external rotation, hip adduction (squeeze test)) measures were investigated using intraclass correlation coefficients (ICC). Reliable hip flexibility and strength measures were compared between football players with (n=10) and without (n=19) groin pain. RESULTS: The bent knee fall out test, hip internal rotation flexibility and the squeeze test demonstrated acceptable (ICC>0.75) intra-rater and inter-rater reliability, while hip external rotation flexibility and hip abduction strength demonstrated acceptable intra-rater but not inter-rater reliability. Hip internal and external rotation strength tests were not found to be reliable. Football players with groin pain had significantly reduced force production on the squeeze test (p>0.05). CONCLUSION: Several hip flexibility and strength measures were found to be reliable. Only the squeeze test discriminated between football players with and without groin pain. PMID- 19282304 TI - Specialisation in sports medicine: the state of the Sport Medicine Specialty Training Core Curriculum in the European Union. PMID- 19282305 TI - Antagonism of the cannabinoid CB-1 receptor protects rat liver against ischaemia reperfusion injury complicated by endotoxaemia. AB - BACKGROUND/AIM: Endotoxaemia can complicate hepatic ischaemia-reperfusion (IR) injury. Endocannabinoids appear to modulate the haemodynamic alterations and cytokine response induced by lipopolysaccharide (LPS). Thus, we aimed to determine the effect of the endocannabinoid CB1-receptor antagonist Rimonabant in a model of hepatic IR injury complicated by endotoxaemia. METHODS: Sprague-Dawley rats pre-treated with Rimonabant 3 or 10 mg/kg or vehicle underwent partial hepatic IR and lipopolysaccharide (LPS) injection at reperfusion. Liver injury was evaluated by serum alanine aminotransferase (ALT) and necrotic-cell count. The inflammatory response was investigated by assessing hepatic neutrophil infiltration, tumour necrosis factor alpha (TNFalpha), interferon gamma (IFNgamma), interleukin 6 (IL6), and suppressor of cytokine signalling (SOCS) 1 and SOCS3 gene expression by real-time polymerase chain reaction (RT-PCR). Systolic blood pressure and hepatic blood flow were measured as haemodynamic parameters. Finally, lipid peroxidation, glutathione status, and immunoreactive CB1 receptor expression in the liver were also determined. RESULTS: Liver injury and neutrophil infiltration occurring in the late-phase of LPS-enhanced IR were significantly reduced by CB1-receptor antagonism. Rimonabant-treated rats showed significantly higher gene expression of IFNgamma, IL6, SOCS1 and SOCS3 in "early" reperfusion, while that of TNFalpha was reduced. These findings were associated with increased STAT3 phosphorylation. Furthermore, CB1-receptor antagonism significantly improved the oxidative injury and haemodynamic alterations occurring during reperfusion in untreated rats. Finally, CB1-receptor immunoreactivity was upregulated early after reperfusion. CONCLUSIONS: This study demonstrates that CB1-receptor antagonism protects the liver against LPS-enhanced IR injury by interfering with the inflammatory response that causes the late, neutrophil-dependent phase of reperfusion injury, although the prevention of the transient endotoxin-related hypotension occurring early during reperfusion may be also involved. PMID- 19282306 TI - Transluminal endoscopic necrosectomy after acute pancreatitis: a multicentre study with long-term follow-up (the GEPARD Study). AB - BACKGROUND: As with endoscopic transmural drainage of peripancreatic fluid collections, the same transluminal access can be expanded to introduce an endoscope through the gastrointestinal wall into the retroperitoneum and remove infected pancreatic necroses under direct visual control. This study reports the first large series with long-term follow-up. METHODS: Data for all patients undergoing transluminal endoscopic removal of (peri)pancreatic necroses between 1999 and 2005 in six different centres were collected retrospectively, and the patients were followed up prospectively until 2008. The initial patient and treatment outcome data were recorded, as were long-term results. RESULTS: Ninety three patients (63 men, 30 women; mean age 57 years) underwent a mean of six interventions starting at a mean of 43 days after an attack of severe acute pancreatitis. After establishment of transluminal access to the necrotic cavity and subsequent endoscopic necrosectomy, initial clinical success was obtained in 80% of the patients, with a 26% complication and a 7.5% mortality rate at 30 days. After a mean follow-up period of 43 months, 84% of the initially successfully treated patients had sustained clinical improvement, with 10% receiving further endoscopic and 4% receiving surgical treatment for recurrent cavities; 16% suffered recurrent pancreatitis. CONCLUSIONS: Direct transluminal endoscopic removal of pancreatic necroses is associated with good long-term maintenance of the high initial efficacy; complications can occur, with an associated mortality of around 7.5%. Further studies are necessary in order to optimise endotherapy and define its role in relation to surgery in the clinical management of such patients. PMID- 19282307 TI - Bcl-2 antagonist killer 1 (BAK1) polymorphisms influence the risk of developing autoimmune rheumatic diseases in women. AB - OBJECTIVE: Bcl-2 antagonist killer 1 (BAK1) is a Bcl-2 family proapoptotic member suggested as a candidate gene for autoimmune diseases. The influence of BAK1 polymorphisms on the risk of developing autoimmune rheumatic diseases (AIRDs) in women was investigated. METHODS: A total of 719 Colombian women were included in the present study: 209 had systemic lupus erythematosus, 99 primary Sjogren syndrome, 159 rheumatoid arthritis and 252 were healthy matched controls. Tag single nucleotide polymorphisms (SNPs) and potentially functional variants were typed by TaqMan allele discrimination assays. HLA-DRB1 and HLA-DQB1 typing was performed by reverse dot-blot hybridisation and linkage disequilibrium (LD) with BAK1 SNPs was assessed. RESULTS: SNPs rs513349 (odds ratio (OR) 0.57, 95% CI 0.46 to 0.72, p = <0.001) and rs5745582 (OR 1.61, 95% CI 1.26 to 2.04, p = <0.001) were associated with the AIRDs included in this study. There was a significant increase of the rs513349G-rs561276C-rs5745582A (GCA) haplotype in each patient cohort as compared to controls (OR 1.95, 95% CI 1.50 to 2.54, p = <0.001). These SNPs were not in LD with HLA-DRB1 or HLA-DQB1 genes. CONCLUSIONS: The results indicate that the BAK1 polymorphisms influence the risk of acquiring AIRDs in the population studied and are consistent with the paradigm that autoimmune diseases are likely to share common susceptibility variants. PMID- 19282308 TI - Women with rheumatoid arthritis negative for anti-cyclic citrullinated peptide and rheumatoid factor are more likely to improve during pregnancy, whereas in autoantibody-positive women autoantibody levels are not influenced by pregnancy. AB - OBJECTIVES: To determine whether changes in levels of anti-cyclic citrullinated peptide (anti-CCP) and rheumatoid factor (RF) are associated with the spontaneous improvement of rheumatoid arthritis (RA) during pregnancy and with the subsequent flare post partum. METHODS: Disease activity scores from the Pregnancy-induced Amelioration of Rheumatoid Arthritis (PARA) study of 118 patients were available for analysis. Before conception (if applicable), at each trimester and at 6, 12 and 26 weeks post partum, levels of the autoantibodies anti-CCP, IgM-RF, IgG-RF and IgA-RF were determined. Responses in disease activity were classified according to European League Against Rheumatism (EULAR) response criteria during pregnancy and post partum, and associated with the presence or absence of autoantibodies. RESULTS: The median levels of anti-CCP and all subclasses of RF during pregnancy were stable, whereas post partum the levels of anti-CCP, IgM-RF and IgA-RF declined. A significantly higher percentage of women without autoantibodies (negative for anti-CCP and RF) improved compared with women positive for either or both autoantibodies (75% vs 39%, p = 0.01). The occurrence of a flare post partum was comparable between these groups. CONCLUSIONS: Improvement of disease activity of RA during pregnancy was not associated with changes in levels of autoantibodies during pregnancy, however, improvement may occur more frequently in the absence of anti-CCP and RF. PMID- 19282309 TI - ASAS/WHO ICF Core Sets for ankylosing spondylitis (AS): how to classify the impact of AS on functioning and health. AB - OBJECTIVE: To report on the results of a standardised consensus process agreeing on concepts typical and/or relevant when classifying functioning and health in patients with ankylosing spondylitis (AS) based on the International Classification of Functioning and Health (ICF). METHODS: Experts in AS from different professional and geographical backgrounds attended a consensus conference and were divided into three working groups. Rheumatologists were selected from members of the Assessment of SpondyloArthritis international Society (ASAS). Other health professionals were recommended by ASAS members. The aim was to compose three working groups with five to seven participants to allow everybody's contribution in the discussions. Experts selected ICF categories that were considered typical and/or relevant for AS during a standardised consensus process by integrating evidence from preceding studies in alternating working group and plenary discussions. A Comprehensive ICF Core Set was selected for the comprehensive classification of functioning and a Brief ICF Core Set for application in trials. RESULTS: The conference was attended by 19 experts from 12 countries. Eighty categories were included in the Comprehensive Core Set, which included 23 Body functions, 19 Body structures, 24 Activities and participation and 14 Environmental factors. Nineteen categories were selected for the Brief Core Set, which included 6 Body functions, 4 Body structures, 7 Activities and participation and 2 Environmental factors. CONCLUSION: The Comprehensive and Brief ICF Core Sets for AS are now available and aim to represent the external reference to define consequences of AS on functioning. PMID- 19282310 TI - Rapid and sustained health utility gain in anti-tumour necrosis factor-treated inflammatory arthritis: observational data during 7 years in southern Sweden. AB - BACKGROUND: Rheumatoid arthritis (RA), psoriatic arthritis (PsA) and other spondylarthritides impose a great impact on the individual in addition to the costs on society, which may be reduced by effective pharmacological treatment. Industry-independent health economic studies should complement studies sponsored by industry. OBJECTIVE: To study secular trends in baseline health utilities in patients commencing tumour necrosis factor (TNF) blockade for arthritis in clinical practice over 7 years; to address utility changes during treatment; to investigate the influence of previous treatment courses; to study the feasibility of health utility measures and to compare them across diagnostic entities. METHODS: EuroQoL 5 dimensions (EQ-5D) utility data were collected from a structured clinical follow-up programme of anti-TNF-treated patients with RA (N = 2554), PsA (N = 574) or spondylarthritides (N = 586). Time trends were calculated. Completer analysis was used. RESULTS: There were weak or non significant secular trends for increasing baseline utilities over time for RA, PsA and spondylarthritides. The maximum gain in utilities had already occurred after 2 weeks for all diagnoses and remained stable for patients remaining on therapy. The first and second anti-TNF courses performed similarly. CONCLUSIONS: Utilities at inclusion remained largely unchanged for RA, PsA and spondylarthritides over 7 years. Improvement occurred early during treatment and not beyond 6 weeks at the group level. Improvement during the first course was not consistently greater than the second. There were no major differences between RA, PsA and spondylarthritides. EQ-5D proved feasible and applicable across these diagnoses. These "real world" data may be useful for health economic modelling. PMID- 19282311 TI - Role of contextual factors in health-related quality of life in ankylosing spondylitis. AB - BACKGROUND: In the bio-psycho-social model of health, the role of contextual factors, either environmental or personal, is recognised. OBJECTIVE: To assess the impact of a number of contextual factors on self-reported disease-specific and generic health-related quality of life in patients with ankylosing spondylitis (AS). METHODS: 522 patients with AS from Canada and Australia completed a postal questionnaire including sociodemographic variables, disease activity (Bath Ankylosing Spondylitis Disease Activity Index (BASDAI)) function (Bath Ankylosing Spondylitis Functional Index (BASFI)) health-related quality of life (ASQoL and EQ-5D) and Rheumatoid Attitudes Index Helplessness Subscale. The contribution of contextual factors (nationality, ethnicity, marital status, education, employment and helplessness) in addition to functioning and disability (BASDAI and BASFI) to health-related quality of life was analysed using multivariate regression analyses. Interactions between contextual variables were explored. RESULTS: Contextual factors explained 37% and 47% of the variance in EQ 5D and ASQoL, respectively. Helplessness and employment were the most important contextual factors. Their role was independent of the strong effect of disease activity (BASDAI) and functional limitations (BASFI). When ASQoL was the outcome, an interaction was seen between employment and education and when EQ-5D was the outcome, an interaction was seen between helplessness and education. CONCLUSIONS: Of the contextual factors explored in this study, helplessness and employment had an important and independent contribution to health-related quality of life. In patients with lower education, the effect of not being employed on ASQoL and the effect of helplessness on EQ-5D were stronger. Contextual factors, especially helplessness and employment, should receive more attention when interpreting data on health-related quality of life. PMID- 19282312 TI - Implantable defibrillators in hypertrophic cardiomyopathy. PMID- 19282313 TI - A randomised, controlled study of rate versus rhythm control in patients with chronic atrial fibrillation and heart failure: (CAFE-II Study). AB - BACKGROUND: Atrial fibrillation (AF) and heart failure (HF) often coexist. The aim was to investigate whether restoring sinus rhythm (SR) could improve cardiac function, symptoms, exercise capacity and quality of life (QoL) in patients with chronic heart failure. METHODS: Patients with HF and persistent AF receiving guideline-recommended treatments, including anticoagulants, were eligible for the study. Patients were randomised to either rhythm (treated with amiodarone for at least 3 months prior to attempting biphasic external cardioversion and continued amiodarone long-term if SR was restored) or rate control. Anticoagulants were continued throughout the study regardless of rhythm, unless contraindications developed. Both groups were treated with beta blockers and/or digoxin to reduce the heart rate to <80 bpm at rest and <110 bpm after walking. Symptoms, walk distance (6-minute corridor walk test, 6MWT), QoL and cardiac function were assessed at baseline and 1 year. RESULTS: 61 patients with HF and persistent AF (median duration 14 months (IQR 5 to 32)) were randomly assigned to a rate or rhythm control strategy. Of patients assigned to rhythm control (n = 30), 66% were in SR at 1 year, and 90% of those assigned to rate control (n = 31) achieved the heart rate target. At 1 year, NYHA class (p = 0.424) and 6MWT distance (p = 0.342) were similar between groups but patients assigned to rhythm control had improved LV function (p = 0.014), NT-proBNP concentration (p = 0.046) and QoL (p = 0.019) compared with those assigned to rate control. Greatest improvement was seen in patients in whom SR was maintained. CONCLUSION: Restoring SR in patients with AF and heart failure may improve QoL and LV function when compared with a strategy of rate control. PMID- 19282314 TI - Device complications and inappropriate implantable cardioverter defibrillator shocks in patients with hypertrophic cardiomyopathy. AB - BACKGROUND: Although the benefit of implantable cardioverter defibrillator (ICD) therapy in patients with hypertrophic cardiomyopathy (HCM) at risk for sudden cardiac arrest is well established, there may be a higher risk for device complications and inappropriate shocks. OBJECTIVES: To determine the incidence of inappropriate ICD shocks and device complications in HCM patients and the impact of young age at ICD implant and atrial fibrillation. METHODS: HCM patients who underwent ICD implantation between January 1988 and December 2005 were included. The frequency of device complications, including pneumothorax, pericardial effusion, haematoma, lead revisions, infection and rate of inappropriate shocks, was determined. ICD shocks were characterised as inappropriate if triggered by sinus tachycardia, atrial fibrillation or device malfunction. RESULTS: A total of 181 patients were included (mean age 44 (SD 17) years; 62% males). During a mean follow-up of 59 (42) months (4.92 years; 830.75 patient-years), 65 patients (36%) had a total of 88 device complications, including 42 (23%) patients with inappropriate shocks. The rate of inappropriate shocks was 5.3% per year (vs 4% risk of appropriate shocks), and the likelihood of inappropriate ICD shocks per 100 patient-years was 5.1. Younger age and atrial fibrillation were associated with an increased risk of inappropriate ICD discharges. CONCLUSIONS: The rate of inappropriate ICD shocks and frequency of device complications in HCM patients are not insignificant and are most common in younger patients and those with atrial fibrillation. Inappropriate ICD shocks are the most common device complication and should be accounted for when counselling high-risk HCM patients for ICD implantation. PMID- 19282315 TI - Association between lung function and disability in African-Americans. AB - BACKGROUND: Impaired lung function is independently associated with higher rates of disability; however, few studies have examined the extent to which this relationship varies by sex. Because men are less likely to have disability, it is expected that the relationship between lung function and disability will be greater among women. METHODS: Logistic regression models were specified to examine the relationship between lung function and disability in 689 African American men and women participating in the Carolina African American Twin Study of Aging. Disability was defined as difficulty in performing at least one of seven basic activities of daily living. Impaired lung function was defined as per cent of predicted peak expiratory flow (PEF) <80. RESULTS: Accounting for demographic and health-related characteristics, women who had impaired lung function had a higher odds (OR 1.77; 95% CI 1.02 to 3.06) of being disabled than those with normal lung function. Impaired lung function was unrelated to disability in men. CONCLUSION: Lung function appears to vary by sex in this sample of African-Americans. Furthermore, among women, lung function should be considered as an important indicator of health. Every effort should be made to improve lung function in African-Americans, but interventions and health promoting strategies may need to be sex specific. PMID- 19282316 TI - Environmental hazards and stress: evidence from the Texas City Stress and Health Study. AB - BACKGROUND: Substantial research has suggested that exposure to environmental health hazards, such as polluting industrial activity, has deleterious effects on psychological and physiological well-being. However, one gap in the existing literature is comparative analysis of objective and subjective exposure's relative association with various measurable outcomes of exposure. METHODS: These relationships were explored within a community sample of 2604 respondents living near a large petrochemical complex in Texas City, Texas, USA. Objective exposure was investigated using distance of residence from a cluster of petrochemical plants and subjective exposure using residents' concern about potential health effects from those plants. Regression models were then used to examine how each type of exposure predicts perceived stress, physiological markers of stress and perceived health. RESULTS: Results suggest that objective exposure was associated primarily with markers of physiological stress (interleukin-6 and viral reactivation), and subjective exposure (concern about petrochemical health risk) was associated with variables assessing perceived health. CONCLUSIONS: From the analysis, it can be inferred that, in the context of an environmental hazard of this type, subjective exposure may be at least as important a predictor of poor health outcomes as objective exposure. PMID- 19282317 TI - Lung cancer mortality and fibre exposures among North Carolina asbestos textile workers. AB - OBJECTIVE: To describe mortality among workers exposed to chrysotile asbestos and evaluate the relationship between lung cancer and asbestos fibre exposure. METHODS: Workers employed for at least 1 day between 1 January 1950 and 31 December 1973 in any of four plants in North Carolina, USA that produced asbestos textile products were enumerated. Vital status was ascertained through 31 December 2003. Historical exposures to asbestos fibres were estimated from work histories and 3578 industrial hygiene measurements taken in 1935-1986. Mortality of the cohort was compared with that of the national population via standardised mortality ratios (SMRs). Exposure-response relationships for lung cancer were examined within the cohort using Poisson regression to compute adjusted mortality rate ratios. RESULTS: Follow-up of 5770 workers included in the cohort resulted in 181 640 person-years of observation, with 2583 deaths from all causes and 277 from lung cancer. Mortality from all causes, all cancers and lung cancer was significant higher than expected, with SMRs of 1.47 for all causes, 1.41 for all cancer and 1.96 (95% CI 1.73 to 2.20) for lung cancer. SMRs for pleural cancer, mesothelioma and pneumoconiosis were also elevated. The risk of lung cancer and asbestosis increased with cumulative fibre exposure (RR 1.102 per 100 fibre year/ml, 95% CI 1.044 to 1.164, and RR 1.249 per 100 fibre-year/ml, 95% CI 1.186 to 1.316, respectively, for total career exposure). CONCLUSIONS: This study provides further evidence that exposure to chrysotile asbestos in textile manufacturing is associated with increased risk of lung cancer, asbestosis cancer of the pleura and mesothelioma. PMID- 19282318 TI - Concordant genotype of upper and lower airways P aeruginosa and S aureus isolates in cystic fibrosis. AB - RATIONALE: Lower airway (LAW) infection with Pseudomonas aeruginosa and Staphylococcus aureus is the leading cause of morbidity in cystic fibrosis (CF). The upper airways (UAW) were shown to be a gateway for acquisition of opportunistic bacteria and to act as a reservoir for them. Therefore, tools for UAW assessment within CF routine care require evaluation. OBJECTIVES: The aims of the study were non-invasive assessment of UAW and LAW microbial colonisation, and genotyping of P aeruginosa and S aureus strains from both segments. METHODS: 182 patients with CF were evaluated (age 0.4-68 years, median 17 years). LAW specimens were preferably sampled as expectorated sputum and UAW specimens by nasal lavage. P aeruginosa and S aureus isolates were typed by informative single nucleotide polymorphisms (SNPs) or by spa typing, respectively. RESULTS: Of the typable S aureus and P aeruginosa isolates from concomitant UAW- and LAW-positive specimens, 31 of 36 patients were carrying identical S aureus spa types and 23 of 24 patients identical P aeruginosa SNP genotypes in both compartments. Detection of S aureus or P aeruginosa in LAW specimens was associated with a 15- or 88-fold higher likelihood also to identify S aureus or P aeruginosa in a UAW specimen from the same patient. CONCLUSIONS: The presence of identical genotypes in UAW and LAW suggests that the UAW play a role as a reservoir of S aureus and P aeruginosa in CF. Nasal lavage appears to be suitable for non-invasive UAW sampling, but further longitudinal analyses and comparison with invasive methods are required. While UAW bacterial colonisation is typically not assessed in regular CF care, the data challenge the need to discuss diagnostic and therapeutic standards for this airway compartment. TRIAL REGISTRATION NUMBER: NCT00266474. PMID- 19282319 TI - Psychometric evaluation of an instrument to assess patient-reported 'psychosocial care by physicians': a structural equation modeling approach. AB - OBJECTIVE: The objective of our study was to develop a theory-based and empirically tested instrument for measuring patient-reported 'psychosocial care by physicians'. We propose a model integrating patients' perceptions with respect to: (i) devotion by physicians, (ii) support by physicians, (iii) information by physicians and (iv) shared decision-making (SDM). DESIGN: Data were gathered during 2001 within a cross-sectional, retrospective mail survey. PARTICIPANTS AND SETTING: A total of 4192 inpatients of six German hospitals. MAIN OUTCOME MEASURE: Specific scales of the Cologne Patient Questionnaire were used. A two step structural equation model procedure was applied. In the first structural equation model, all items were modeled as indicators of the intended underlying latent construct, 'psychosocial care by physicians'. In the second structural equation model, criterion-related validity of the intended construct was tested with respect to patients' 'satisfaction', 'trust in physicians' and the 'image of the hospital'. RESULTS: The results confirmed that the aspects of psychosocial care provided by physicians measured by the scale items are indeed indicators of the same construct. Furthermore, indicator reliabilities and selectivities revealed that the content of all 13 items was highly representative of the underlying construct. The second structural equation model showed that 'psychosocial care by physicians' is related to 'patients' satisfaction', 'trust in physicians' and 'hospital-image' in a significant and relevant manner. CONCLUSION: On the basis of our instrument's reported psychometric characteristics and of the initial validity indicators, it may be regarded as an adequate measure for further use in outcome and intervention research, and as a quality indicator for the physician-patient relationship. PMID- 19282320 TI - Flower thermoregulation facilitates fertilization in Asian sacred lotus. AB - BACKGROUND AND AIMS: The thermoregulatory flower of the Asian sacred lotus (Nelumbo nucifera) can maintain a relatively stable temperature despite great variations in ambient temperature during anthesis. The thermoregulation has been hypothesized to offer a direct energy reward for pollinators in lotus flowers. This study aims to examine whether the stable temperature maintained in the floral chamber influences the fertilization process and seed development. METHODS: An artificial refrigeration instrument was employed to cool flowers during the fertilization process and post-fertilization period in an experimental population. The effect of temperature on post-pollination events was also examined by removing petals in two field populations. KEY RESULTS: Treatments with low floral temperature did not reduce stigma receptivity or pollen viability in undehisced anthers. Low temperature during the fertilization period significantly decreased seed set per flower but low temperature during the phase of seed development had no effect, suggesting that temperature regulation by lotus flowers facilitated fertilization success. Hand-pollination treatments in two field populations indicated that seed set of flowers with petals removed was lower than that of intact flowers in north China, where ambient temperatures are low, but not in south China, confirming that reducing the temperature of carpels did influence post-pollination events. CONCLUSIONS: The experiments suggest that floral thermoregulation in lotus could enhance female reproductive success by facilitating fertilization. PMID- 19282321 TI - A new training set-up for trans-apical aortic valve replacement. AB - Trans-apical aortic valve replacement (AVR) is a new and rapidly growing therapy. However, there are only few training opportunities. The objective of our work is to build an appropriate artificial model of the heart that can replace the use of animals for surgical training in trans-apical AVR procedures. To reduce the necessity for fluoroscopy, we pursued the goal of building a translucent model of the heart that has nature-like dimensions. A simplified 3D model of a human heart with its aortic root was created in silico using the SolidWorks Computer-Aided Design (CAD) program. This heart model was printed using a rapid prototyping system developed by the Fab@Home project and dip-coated two times with dispersion silicone. The translucency of the heart model allows the perception of the deployment area of the valved-stent without using heavy imaging support. The final model was then placed in a human manikin for surgical training on trans apical AVR procedure. Trans-apical AVR with all the necessary steps (puncture, wiring, catheterization, ballooning etc.) can be realized repeatedly in this setting. PMID- 19282322 TI - Acute thrombosis after endarterectomy of stented left anterior descending artery. AB - Acute thrombosis of the endarterectomized coronary artery is a serious complication after coronary endarterectomy. Herein, we describe a case of a 65 year-old man who had undergone percutaneous stent implantation in the left anterior descending artery, then after he received an endarterectomy with the removal of stents severe in-stent restenosis occurred. Three days after the operation, sick sinus syndrome developed with acute myocardial infarction. Coronary angiography revealed thrombosis at the reconstructed site of the left anterior descending artery. Pacemaker implantation, intra-aortic balloon pumping, and aggressive anticoagulation produced recanalization of the left anterior descending artery. PMID- 19282323 TI - Drug uptake in a rodent sarcoma model after intravenous injection or isolated lung perfusion of free/liposomal doxorubicin. AB - The distribution of free and liposomal doxorubicin (Liporubicin) administered by intravenous injection (IV) or isolated lung perfusion (ILP) was compared in normal and tumor tissues of sarcoma bearing rodent lungs. A single sarcomatous tumor was generated in the left lung of 35 Fischer rats, followed 10 days later by left-sided ILP (n=20) or IV drug administration (n=12), using 100 microg and 400 microg free or liposomal doxorubicin, respectively. The tumor and lung tissue drug concentration was measured by HPLC. Free doxorubicin administered by ILP resulted in a three-fold (100 microg) and 10-fold (400 microg) increase of the drug concentration in the tumor and normal lung tissue compared to IV administration. In contrast, ILP with Liporubicin resulted in a similar drug uptake in the tumor and lung tissue compared to IV injection. For both drug formulations and dosages, ILP resulted in a higher tumor to lung tissue drug ratio but also in a higher spatial heterogeneity of drug distribution within the lung compared to IV administration. ILP resulted in a higher tumor to lung tissue drug ratio and in a more heterogeneous drug distribution within the lung compared to IV drug administration. PMID- 19282324 TI - Systemic lupus erythematosus with multiple lung cysts. AB - We report a rare case of recurrent pneumothorax complicated by systemic lupus erythematosus (SLE) with multiple lung cysts. The patient was a 53-year-old woman without a history of smoking. At the age of 29, she was diagnosed with SLE. A computed tomography (CT) scan of the chest showed multiple, bilateral lung cysts. She had recurrent episodes of spontaneous pneumothorax on the same side within a period of two months. Although pulmonary involvement is common in SLE patients, cystic lung disease associated with SLE is extremely rare. PMID- 19282325 TI - A re-examination of proliferation and differentiation of type A spermatogonia in the adult rhesus monkey (Macaca mulatta). AB - BACKGROUND: Companion studies using an experimental non-human primate paradigm known as a testicular clamp indicated that the behavior of undifferentiated type A spermatogonia did not conform fully to earlier classical models. This issue was therefore re-examined in normal monkeys. METHODS: Adult male rhesus monkeys (n = 4) received an i.v. bolus of 5-bromo-2'-deoxyuridine (BrdU): one testis (first) was removed 3 h later and the remaining testis (second) was removed after 11 days and 3 h. Tissue was fixed in Bouin's solution, and numbers of A dark (Ad), small A pale (Aps) and large A pale spermatogonia, differentiating B spermatogonia, S phase-labeled and degenerating cells were enumerated. Data are given as mean +/- SEM. RESULTS: During the early stages of the seminiferous epithelial cycle in the first testis, Ap spermatogonia (1.3 cells/cross section) were predominantly Aps (nuclear dia., 7.1 +/- 0.1 microm). Aps were never S-phase labeled. Apl (nuclear dia., 8.8 +/- 0.5 microm) appeared in Stages IV-VI and were maximal in Stages VII X when S-phase labeling of this phenotype at 3 h was greatest. The first generation of B spermatogonia appeared in Stages XI-XII (0.84 cells/cross section). Using cells/cross section, the ratio of Ap (Stages I V):B1:B2:B3:B4:preleptotene spermatocyte was 1:0.7:1.4:2.8:5.6:11.2. In the second testis, labeled Aps (and Apl) were observed. Ad were not BrdU labeled, and degenerating cells were rarely observed. CONCLUSIONS: The results are not entirely consistent with earlier models of spermatogonial proliferation and differentiation in the monkey. Most notably, our findings suggest that in any one cycle of the seminiferous epithelium only a fraction of Ap spermatogonia is mitotically active. PMID- 19282326 TI - GPR119 is required for physiological regulation of glucagon-like peptide-1 secretion but not for metabolic homeostasis. AB - G protein-coupled receptor 119 (GPR119) is expressed in pancreatic islets and intestine, and is involved in insulin and incretin hormone release. GPR119 knockout (Gpr119(-/-)) mice were reported to have normal islet morphology and normal size, body weight (BW), and fed/fasted glucose levels. However, the physiological function of GPR119 and its role in maintaining glucose homeostasis under metabolic stress remain unknown. Here, we report the phenotypes of an independently generated line of Gpr119(-/-) mice under basal and high-fat diet (HFD)-induced obesity. Under low-fat diet feeding, Gpr119(-/-) mice show normal plasma glucose and lipids, but have lower BWs and lower post-prandial levels of active glucagon-like peptide 1 (GLP-1). Nutrient-stimulated GLP-1 release is attenuated in Gpr119(-/-) mice, suggesting that GPR119 plays a role in physiological regulation of GLP-1 secretion. Under HFD-feeding, both Gpr119(+)(/)(+) and Gpr119(-/-) mice gain weight similarly, develop hyperinsulinemia and hyperleptinemia, but not hyperglycemia or dyslipidemia. Glucose and insulin tolerance tests did not reveal a genotypic difference. These data show that GPR119 is not essential for the maintenance of glucose homeostasis. Moreover, we found that oleoylethanolamide (OEA), reported as a ligand for GPR119, was able to suppress food intake in both Gpr119(+)(/)(+) and Gpr119(-/-) mice, indicating that GPR119 is not required for the hypophagic effect of OEA. Our results demonstrate that GPR119 is important for incretin and insulin secretion, but not for appetite suppression. PMID- 19282328 TI - Vinaxanthone, a new FabI inhibitor from Penicillium sp. AB - OBJECTIVES: Bacterial enoyl-ACP reductase (FabI) has been validated as a novel antibacterial target for tackling infections caused by multidrug-resistant pathogens. A few FabI inhibitors, however, are known. This study isolated a new FabI inhibitor from Penicillium sp. METHODS: A screening programme led to the selection of a Penicillium sp. producing a strong FabI-inhibitory metabolite. The chemical structure of the isolated FabI inhibitor was elucidated by mass spectrometry and nuclear magnetic resonance spectral data. The antibacterial target of the inhibitor was validated by overexpression assays. RESULTS: The isolated FabI inhibitor was elucidated to be vinaxanthone. It selectively inhibited Staphylococcus aureus FabI with an IC(50) of 0.9 microM; it did not affect FabK, an enoyl-ACP reductase of Streptococcus pneumoniae. Consistent with its inhibition of FabI, the inhibitor prevented intracellular fatty acid synthesis while it did not affect protein biosynthesis. It also prevented the growth of S. aureus as well as methicillin-resistant S. aureus (MRSA) and quinolone-resistant S. aureus. Importantly, fabI-overexpressing S. aureus showed reduced susceptibility to the inhibitor compared with the wild-type strain, demonstrating that its antibacterial action is mediated by inhibition of FabI. CONCLUSIONS: Vinaxanthone is a new FabI-directed antibacterial of natural origin that could have potential for further development as a new anti-MRSA agent. PMID- 19282327 TI - Endometrial cysteine-rich secretory protein 3 is inhibited by human chorionic gonadotrophin, and is increased in the decidua of tubal ectopic pregnancy. AB - Ectopic pregnancy (EP) remains a considerable cause of morbidity and occasional mortality. Currently, there is no reliable test to differentiate ectopic from intrauterine gestation. We have previously used array technology to demonstrate that differences in gene expression in decidualized endometrium from women with ectopic and intrauterine gestations could be used to identify candidate diagnostic biomarkers for EP. The aim of this study was to further investigate the decidual gene with the highest fold increase in EP, cysteine-rich secretory protein-3 (CRISP-3). Decidualized endometrium from gestation-matched women undergoing surgical termination of pregnancy (n = 8), evacuation of uterus for miscarriage (n = 6) and surgery for EP (n = 11) was subjected to quantitative RT PCR, morphological assessment, immunohistochemistry and western blot analysis. Sera were analysed for progesterone and human chorionic gonadotrophin (hCG) levels. Immortalized endometrial epithelial cells were cultured with physiological concentrations of hCG. CRISP-3 mRNA and protein expression were greater in endometrium from ectopic when compared with intrauterine pregnancies (P < 0.05). CRISP-3 protein was localized to epithelium and granulocytes of endometrium. CRISP-3 serum concentrations were not different in women with ectopic compared with intrauterine pregnancies. CRISP-3 expression in endometrium was not related to the degree of decidualization or to serum progesterone levels. Endometrial CRISP-3 expression was inversely proportional to serum hCG concentrations (P < 0.001). Stimulation of endometrial epithelial cells with hCG in vitro caused a reduction in CRISP-3 expression (P < 0.01). The measurement of CRISP-3 in endometrium could provide an additional tool in the diagnosis of failing early pregnancy of unknown location. The absence of a local reduction in expression of CRISP-3 in decidualized endometrium of women with EP may be due to reduced exposure to hCG due to the ectopic location of the trophoblast. PMID- 19282330 TI - Effect of a solution containing citrate/Methylene Blue/parabens on Staphylococcus aureus bacteria and biofilm, and comparison with various heparin solutions. AB - OBJECTIVES: Some antibiotic solutions increase bacterial resistance and may cause toxic side effects. Heparin, frequently used as an anticoagulant in catheter lock solutions, may cause bleeding and stimulate biofilm formation. The aim of this study was to investigate the effect of a new antibacterial/antithrombotic solution, citrate/Methylene Blue/parabens (C/MB/P), versus various heparin solutions on the viability and the structure of preformed mature biofilms of Staphylococcus aureus bacteria. The degree of eradication of both planktonic and sessile microorganisms was evaluated. METHODS: The changes in the structure of biofilms after exposure to C/MB/P and several concentrations of heparin were analysed by means of confocal laser scanning microscopy. COMSTAT image analysis was utilized to compare biofilm biomass, average and maximum height, surface coverage and roughness coefficient. Viability studies were performed on both biofilms and supernatant solutions. RESULTS: C/MB/P, in contrast to heparin solutions, significantly reduced biofilm biomass and thickness and reduced viability by 5 log when compared with saline treatment. No viable planktonic bacteria were detected and the few remaining biofilm cells appeared to be lysed. In contrast, most heparin solutions only reduced viability up to 1.0 log and failed to eradicate planktonic bacteria. CONCLUSIONS: C/MB/P has a rapid bactericidal effect on the preformed, mature biofilm of S. aureus. The structural changes of biofilms treated with C/MB/P, together with the observed log reduction of viable biofilm cells, confirmed the high potential of this solution to eliminate sessile bacteria. Furthermore, the tested solution entirely eliminated planktonic bacteria detached from the biofilm. PMID- 19282331 TI - Guidelines (2008) for the prophylaxis and treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections in the United Kingdom. AB - These evidence-based guidelines are an updated version of those published in 2006. They have been produced after a literature review of the treatment and prophylaxis of methicillin-resistant Staphylococcus aureus (MRSA). The guidelines aim to complement those recently published for the antibiotic treatment of common and emerging community-onset MRSA infections in the UK. The guidelines have reviewed and updated, where appropriate, previous recommendations, taking into account any changes in the UK epidemiology of MRSA, ongoing national surveillance data and the value of new antistaphylococcal agents licensed for use in UK practice. Emerging therapies that have not been licensed for UK use are not reviewed, but their future potential role has been mentioned where deemed appropriate. Recommendations are given for the treatment of common infections caused by MRSA, elimination of MRSA from carriage sites and prophylaxis of surgical site infection. PMID- 19282332 TI - The potential role of snus products within a tobacco harm reduction strategy. PMID- 19282333 TI - Measles outbreak in Qassim, Saudi Arabia. PMID- 19282335 TI - Prospective parental consent for autopsy research following sudden unexpected childhood deaths: a successful model. AB - BACKGROUND: Organ retention issues, recent changes in the Coroners' (Amendment) Rules 2005 and the Human Tissue Act have resulted in pessimism regarding prospective consent for paediatric autopsy research in the UK. OBJECTIVES: To examine the feasibility and acceptability of a prospective telephone consenting model for post-mortem magnetic resonance (MR) imaging research in HM Coroners' cases. DESIGN: Following each autopsy referral from the HM Coroner, permission to contact the family for research was requested. A family liaison sister, with experience in dealing with bereaved families, then contacted the parents by telephone, explained the study and obtained oral, and then written consent for post-mortem imaging. SETTING: London and an area south of London. RESULTS: Of 76 eligible HM Coroners' cases referred during the study period, permission to contact parents (provided by the HM Coroners' Office) was obtained for only 32 cases (42%). The research sister contacted 32 parents during the study period of whom 31 (96.8%) gave oral research consent. "Helping other parents in the future" and "the importance of post-mortem research" were the main reasons for parents wanting to participate in research. CONCLUSIONS: Prospective consenting for HM Coroners' cases for research is feasible in children, and can be done ethically by parental consenting via telephone contact before autopsy by appropriately trained staff. However, close co-ordination between mortuary staff, HM Coroners, research staff and medical staff is required. This model may be useful in performing post-mortem research in HM Coroners' cases and in developing paediatric tissue and brain banks in the UK. PMID- 19282336 TI - Academic attainment and special educational needs in extremely preterm children at 11 years of age: the EPICure study. AB - AIM: To assess academic attainment and special educational needs (SEN) in extremely preterm children in middle childhood. METHODS: Of 307 extremely preterm (< or =25 weeks) survivors born in the UK and Ireland in 1995, 219 (71%) were re assessed at 11 years of age and compared to 153 classmates born at term, using standardised tests of cognitive ability and academic attainment and teacher reports of school performance and SEN. Multiple imputation was used to correct for selective dropout. RESULTS: Extremely preterm children had significantly lower scores than classmates for cognitive ability (-20 points; 95% CI -23 to 17), reading (-18 points; -22 to -15) and mathematics (-27 points; -31 to -23). Twenty nine (13%) extremely preterm children attended special school. In mainstream schools, 105 (57%) extremely preterm children had SEN (OR 10; 6 to 18) and 103 (55%) required SEN resource provision (OR 10; 6 to 18). Teachers rated 50% of extremely preterm children as having below average attainment compared with 5% of classmates (OR 18; 8 to 41). Extremely preterm children who entered compulsory education an academic year early due to preterm birth had similar academic attainment but required more SEN support (OR 2; 1.0 to 3.6). CONCLUSIONS: Extremely preterm survivors remain at high risk for learning impairments and poor academic attainment in middle childhood. A significant proportion require full-time specialist education and over half of those attending mainstream schools require additional health or educational resources to access the national curriculum. The prevalence and impact of SEN are likely to increase as these children approach the transition to secondary school. PMID- 19282337 TI - Human salt taste and the lingual surface potential correlate. AB - We have demonstrated in humans that Na(+) evokes changes in the lingual surface potential (LSP) using a custom chamber. To assess whether a relationship exists between the Na(+)-evoked changes in the LSP and the intensity of salt taste, we measured the LSP and the intensity of salt taste simultaneously in 7 subjects using test solutions (50, 100, 300, and 1000 mM NaCl) presented in random order. The evoked LSPs and intensity scores correlated with one another well (r(2) = 0.992, P < 0.01). We then screened 14 subjects for their ability to discriminate between 100 and 300 mM NaCl using the chamber. Three subjects were consistently capable of distinguishing the salt concentrations. In these 3 subjects, an inhibitor of the epithelial sodium channel, amiloride (10 muM), blocked the ability to distinguish salt concentrations and affected the LSP. These data suggest that the LSP may be a component of the signal transduction system involved in human salt taste. In adept salt tasters, an amiloride-sensitive mechanism appears to have a role in distinguishing salt concentrations. PMID- 19282340 TI - Airway smooth muscle thickness in asthma is related to severity but not duration of asthma. AB - Asthma is characterised by an increased airway smooth muscle (ASM) area (ASM(area)) within the airway wall. The present study examined the relationship of factors including severity and duration of asthma to ASM(area). The perimeter of the basement membrane (PBM) and ASM(area) were measured on transverse sections of large and small airways from post mortem cases of fatal (n = 107) and nonfatal asthma (n = 37) and from control subjects (n = 69). The thickness of ASM (ASM(area)/PBM) was compared between asthma groups using multivariate linear regression. When all airways were considered together, ASM(area)/PBM (in millimetres) was increased in nonfatal (median 0.04; interquartile range 0.013 0.051; p = 0.034) and fatal cases of asthma (0.048; 0.025-0.078; p<0.001) compared with controls (0.036; 0.024-0.042). Compared with cases of nonfatal asthma, ASM(area)/PBM was greater in cases of fatal asthma in large (p<0.001) and medium (p<0.001), but not small, airways. ASM(area)/PBM was not related to duration of asthma, age of onset of asthma, sex or smoking. No effect due to study centre, other than that due to sampling strategy, was found. The thickness of the ASM layer is increased in asthma and is related to the severity of asthma but not its duration. PMID- 19282341 TI - Pulmonary function and exercise capacity in survivors of congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is associated with pulmonary hypoplasia and pulmonary hypertension. The objective of this study was to assess pulmonary function and exercise capacity and its early determinants in children and adolescents born with high-risk CDH (CDH-associated respiratory distress within the first 24 h) and to explore the relationship of these findings with CDH severity. Of 159 patients born with high-risk CDH, 84 survived. Of the 69 eligible patients, 53 children (mean+/-SD age 11.9+/-3.5 yrs) underwent spirometry, lung volume measurements and maximal cardiopulmonary exercise testing (CPET). Results of the pulmonary function tests were compared with those from a healthy control group matched for sex, age and height. CDH survivors had a significantly lower forced expiratory volume in 1 s (FEV(1)), forced vital capacity (FVC), FEV(1)/FVC, maximum mid-expiratory flow and peak expiratory flow when compared with healthy controls. The residual volume/total lung capacity ratio was significantly higher. Linear regression analysis showed that gastro oesophageal reflux disease was an independent determinant of reduced FEV(1) and FVC. CPET results were normal in those tested. High-risk CDH survivors have mild to moderate pulmonary function abnormalities when compared with a healthy matched control group, which may be related to gastro-oesophageal reflux disease in early life. Exercise capacity and gas exchange parameters were normal in those tested, indicating that the majority of patients do not have physical impairment. PMID- 19282342 TI - Reliability of ventilatory parameters during cycle ergometry in multicentre trials in COPD. AB - We studied the distribution profiles and repeatability of key exercise performance parameters in the first large multicentre trials to include these measurements in chronic obstructive pulmonary disease (COPD). After a screening visit, 463 subjects with COPD (mean+/-SD forced expiratory volume in 1 s 43+/-13% predicted) completed two run-in visits before treatment randomisation. At the run in visits, measurements were conducted at rest, at a standardised time near end exercise (isotime) and at peak exercise during constant work rate (CWR) cycle tests at 75% of each individual's maximum work capacity. The intraclass correlation coefficient was used to evaluate the test-retest repeatability of measurements of endurance time (ET), inspiratory capacity (IC), ventilation and dyspnoea intensity (Borg scale) during exercise. IC, ventilation and dyspnoea ratings were normally distributed; ET showed rightward skew (median or = 0.87). Ventilation was repeatable over the same time-points (R > or = 0.92), as was dyspnoea intensity at isotime (R = 0.79) and at peak exercise (R = 0.81). In conclusion, key perceptual and ventilatory parameters can be reliably measured during CWR cycle exercise in multicentre clinical trials in moderate to very severe COPD. PMID- 19282343 TI - Mature CD11c(+) cells are enhanced in hypersensitivity pneumonitis. AB - The present study verified the hypothesis that enhanced maturation of antigen presenting CD11c(+) cells could explain the viral-induced exacerbated immune response to Saccharopolyspora rectivirgula (SR), the main antigen responsible for farmer's lung, a classic form of hypersensitivity pneumonitis (HP). Four groups of mice were studied: group 1 received intranasal instillations of saline; group 2 received instillations of SR for 12 weeks; group 3 received instillations of saline and a single infection with Sendai virus on week 3; and group 4 received instillations of SR for 12 weeks with a single administration of Sendai virus on week 3. On week 13, mice were sacrificed and bronchoalveolar lavage was performed. Lungs were harvested, digested with enzymes, and CD11c(+) cells were analysed in flow cytometry with anti-CD11c, anti-CD86 and anti-major histocompatibility complex class II markers. Immunofluorescence studies were also performed with the same cell surface markers. Both flow cytometry and immunofluorescence results demonstrate that mature CD11c(+) cells are significantly enhanced in SR-challenged mice simultaneously infected with Sendai virus, compared with other groups. These CD11c(+) cells persist in the lung for 9 weeks after the virus infection. Maturation of CD11c(+) cells could explain, at least in part, the virus-induced increased immune response to SR antigens in this model of HP, but mechanisms have still to be elucidated. PMID- 19282344 TI - Pulmonary arterial hypertension in limited cutaneous systemic sclerosis: a distinctive vasculopathy. AB - Systemic sclerosis-associated pulmonary arterial hypertension (SScPAH) has a worse prognosis and response to pulmonary arterial hypertension (PAH) therapy than idiopathic PAH (IPAH). These differences have not yet been explained. Knowledge concerning histological pulmonary vasculopathy in SScPAH is limited in contrast to IPAH. Therefore, we explored patterns of vasculopathy in SScPAH compared with IPAH. Parameters of vasculopathy were assessed from lung tissue of eight PAH patients with limited cutaneous systemic sclerosis and 11 IPAH patients. Lung tissue was obtained at autopsy (n = 15), explantation (n = 3) and biopsy (n = 1). Pulmonary arterial/arteriolar intimal fibrosis was identified in all SScPAH patients and in three IPAH patients (p = 0.003). Fibrosis of pulmonary veins/venules was found in all SScPAH patients and in three IPAH patients (p = 0.003). In four SScPAH patients, fibrosis of veins/venules was focal and associated with capillary congestion as in pulmonary veno-occlusive disease (PVOD). Of the IPAH patients, 10 had unequivocal evidence of plexogenic arteriopathy compared with none of the SScPAH patients (p = 0.001). SScPAH is characterised by small vessel intimal fibrosis, which is associated with a PVOD like pattern in some cases. This might explain its different clinical behaviour from IPAH. Small vessel intimal fibrosis may provide clues to elucidation of differences in pathogenetic mechanisms between the groups. PMID- 19282345 TI - Informed participation in a randomised controlled trial of computed tomography screening for lung cancer. AB - The actual lung cancer (screening) knowledge, attitudes, risk perceptions, reasons to participate in or decline participation, and informed decisions of subjects who decided to or decided not to participate in the Dutch-Belgian randomised controlled trial for lung cancer screening in high-risk subjects (the NELSON trial) were evaluated. A total of 2,500 high-risk subjects were asked to complete a questionnaire 3 weeks after they had received a brochure with information about the trial. Differences in knowledge, attitude and risk perception between participants and nonparticipants were analysed with logistic regression analyses adjusted for sex and smoking status. The questionnaire response of trial participants was 80% (n = 889) whereas the response of nonparticipants was low (7%, n = 97) and selective. Participants' responses to knowledge items on lung cancer as a disease were on average more often correct (mean+/-sd 68+/-17%) than items on lung cancer screening (49+/-29%). Participants had adequate knowledge on lung cancer screening (51%) more often than the nonparticipants (38%; p = 0.009). Of the decisions regarding participation, 49% were uninformed, mainly due to insufficient knowledge. Most of the participants (99%) and 64% of the nonparticipants had a positive attitude towards lung cancer screening. Additional efforts are required to improve the knowledge and understanding of subjects who are in the process of decision-making regarding participation in a lung cancer screening trial. PMID- 19282346 TI - Decreased hyaluronan in airway smooth muscle cells from patients with asthma and COPD. AB - Glycosaminoglycans (GAG) are essential extracellular matrix molecules which regulate tissue flexibility, a parameter that is reduced in airways of patients with asthma and chronic obstructive pulmonary disease (COPD). We investigated the expression of GAG and their metabolising enzymes in primary human airway smooth muscle cells (ASMC) obtained from healthy donors (controls) and patients with asthma or COPD. Total GAG synthesis was assessed by [(3)H]-glucosamine incorporation. GAG were isolated, purified, fractionated by electrophoresis and characterised using specific GAG-degrading enzymes. Secretion of hyaluronic acid (HA) by ASMC from patients with asthma or COPD was significantly decreased compared with controls. RT-PCR analysis and western blotting revealed that this decrease was associated with a significant reduction in the expression of HA synthase-1 and -2 and a significant increase of hyaluronidase-1. Furthermore, the expression of the HA receptor CD44 was significantly decreased, whereas the receptor for HA-mediated motility was not expressed in asthma or COPD. Our results indicate that there is a decreased expression of HA in asthma and COPD associated with a synergistic regulation of HA metabolising enzymes that may regulate the pathological airway remodelling in these lung diseases. PMID- 19282347 TI - Physiological responses to linear treadmill and cycle ergometer exercise in COPD. AB - Incremental cardiopulmonary exercise testing work rate ideally increases linearly to the subject's tolerance within approximately 10 min. Widely used treadmill protocols often yield shorter exercise times in debilitated patients. We compared a recently described treadmill protocol featuring linear work rate increase, weight adjustments and a priori exercise tolerance estimates with standard cycle and treadmill protocols. We also compared treadmill and cycle responses to examine mechanisms of oxyhaemoglobin desaturation differences. In total, 16 subjects with chronic obstructive pulmonary disease (COPD; mean+/-sd forced expiratory volume in 1 s of 36.5+/-10.9% predicted) performed incremental exercise using cycle, linear treadmill and modified Bruce protocols. Initial linear treadmill speed and grade yielded oxygen uptake (V'(O(2))) similar to cycle unloaded pedalling; Bruce protocol first stage elicited much higher V'(O(2)). Exercise duration was much shorter in Bruce than in cycle or linear treadmill protocols. At peak exercise, greater desaturation was noted in linear treadmill and Bruce protocols compared with cycle (-8.9+/-4.9 versus -8.5+/-4.7 versus -3.7+/-3.3%; p<0.001); at iso-V'(O(2)) values this difference widened as exercise proceeded. Iso-V'(O(2)) desaturation differences were largely related to higher ventilatory response to cycle than to treadmill exercise. The linear incremental treadmill protocol generates responses similar to cycle ergometry in severe COPD. However, cycle ergometry elicits less desaturation than does ambulation, making the linear treadmill protocol advantageous when evaluating COPD patients. PMID- 19282348 TI - A retrospective TBNET assessment of linezolid safety, tolerability and efficacy in multidrug-resistant tuberculosis. AB - Linezolid is used to treat patients with multidrug-resistant (MDR)/extensively drug-resistant (XDR)-tuberculosis (TB) cases, although clinical data on its safety, tolerability and efficacy are lacking. We performed a retrospective, nonrandomised, unblinded observational study evaluating the safety and tolerability of linezolid at 600 mg q.d. or b.i.d. in MDR/XDR-TB treatment in four European countries. Efficacy evaluation compared end-points of 45 linezolid treated against 110 linezolid-nontreated cases. Out of 195 MDR/XDR-TB patients, 85 were treated with linezolid for a mean of 221 days. Of these, 35 (41.2%) out of 85 experienced major side-effects attributed to linezolid (anaemia, thrombocytopenia and/or polyneuropathy), requiring discontinuation in 27 (77%) cases. Most side-effects occurred after 60 days of treatment. Twice-daily administration produced more major side-effects than once-daily dosing (p = 0.0004), with no difference in efficacy found. Outcomes were similar in patients treated with/without linezolid (p = 0.8), although linezolid-treated cases had more first-line (p = 0.002) and second-line (p = 0.02) drug resistance and a higher number of previous treatment regimens (4.5 versus 2.3; p = 0.07). Linezolid 600 mg q.d. added to an individualised multidrug regimen may improve the chance of bacteriological conversion, providing a better chance of treatment success in only the most complicated MDR/XDR-TB cases. Its safety profile does not warrant use in cases for which there are other, safer, alternatives. PMID- 19282349 TI - Acute myelogenous leukemia in older adults. AB - The incidence of acute myelogenous leukemia (AML) increases with age. Older AML patients, generally defined by age > or = 60 years, have worse treatment outcomes than younger patients. While selected older patients can benefit from standard therapies, as a group they experience greater treatment-related toxicity, lower remission rates, shorter disease-free survival times, and shorter overall survival times. Outcome disparity is in part explained by age-related biologic features. Older patients are more likely to present with unfavorable cytogenetic abnormalities, multidrug resistance phenotypes, and secondary AML. However, even older adults with favorable tumor biology have a worse prognosis than younger patients. Patient-specific factors, including impaired physical function and comorbidity, independently predict greater treatment toxicity and shorter survival. Improving patient assessment strategies is critical to identify those patients who are most likely to benefit from induction and postremission therapies. In addition, continued efforts to identify more effective and tolerable induction and postremission strategies are needed for this population. Investigations of hypomethylating agents and signal transduction inhibitors hold promise for the treatment of AML patients. Steady advances in the field of hematopoietic transplantation, including use of reduced intensity transplants, may result in additional curative options available to selected older adults. Finally, improved supportive care strategies are needed to maximize treatment outcomes. PMID- 19282350 TI - FDA review of a panitumumab (Vectibix) clinical trial for first-line treatment of metastatic colorectal cancer. AB - On September 27, 2006, the U.S. Food and Drug Administration granted accelerated approval to panitumumab (Vectibix; Amgen, Inc., Thousand Oaks, CA) for the treatment of patients with epidermal growth factor receptor-expressing, metastatic colorectal carcinoma with disease progression on or following fluoropyrimidine-, oxaliplatin-, and irinotecan-containing chemotherapy regimens. Accelerated approval was based on demonstration of a beneficial effect on progression-free survival (PFS). The present submission summarizes a second clinical trial, to be included in the panitumumab package insert in June 2008, of chemotherapy and bevacizumab with and without panitumumab in the first-line treatment of patients with metastatic colorectal cancer. The study was closed when inferior PFS and greater toxicity were demonstrated at the time of the planned interim efficacy analysis. Patients receiving panitumumab in combination with bevacizumab and chemotherapy experienced a higher incidence of death (9% versus 4%) and a higher risk for grade 3 and 4 toxicities than patients receiving bevacizumab and chemotherapy alone. The incidences of any Common Terminology Criteria for Adverse Events grade 3 and 4 adverse events (AEs) were 87% and 72% in the panitumumab and control groups, respectively. Grade 3 and 4 AEs occurring more commonly in panitumumab-treated patients included rash/acneiform dermatitis, diarrhea, dehydration, primarily resulting from diarrhea, hypokalemia, stomatitis/mucositis, and pulmonary embolism. The addition of panitumumab to bevacizumab and chemotherapy for the first-line treatment of metastatic colorectal cancer was harmful when compared with bevacizumab and chemotherapy alone. The use of panitumumab in this setting cannot be recommended. PMID- 19282351 TI - Sphingosine-1-phosphate receptor signalling in the heart. AB - The five known members of the sphingosine-1-phosphate (S1P) receptor family exhibit diverse tissue expression profiles and couple to distinct G-protein mediated signalling pathways. S1P1, S1P2, and S1P3 receptors are all present in the heart, but the ratio of these subtypes differs for various cardiac cells. The goal of this review is to summarize data concerning which S1P receptor subtypes regulate cardiac physiology and pathophysiology, which G-proteins and signalling pathways they couple to, and in which cell types they are expressed. The available information is based on studies using a lamentably limited set of pharmacological agonists/antagonists, but is complemented by work with S1P receptor subtype-specific knockout mice and sphingosine kinase knockout mice. In cardiac myocytes, the S1P1 receptor subtype is the predominant subtype expressed, and the activation of this receptor inhibits cAMP formation and antagonizes adrenergic receptor-mediated contractility. The S1P3 receptor, while expressed at lower levels, mediates the bradycardic effect of S1P agonists. Studies using knockout mice indicate that S1P2 and S1P3 receptors play a major role in mediating cardioprotection from ischaemia/reperfusion injury in vivo. S1P receptors are also involved in remodelling, proliferation, and differentiation of cardiac fibroblasts, a cell type in which the S1P3 receptor predominates. Receptors for S1P are also present in endothelial and smooth muscle cells where they mediate peripheral vascular tone and endothelial responses, but the role of this regulatory system in the cardiac vasculature is unknown. Further understanding of the contributions of each cell and receptor subtype to cardiac function and pathophysiology should expedite consideration of the endogenous S1P signalling pathway as a therapeutic target for cardiovascular disease. PMID- 19282352 TI - Stabilization of glucose in blood samples: why it matters. PMID- 19282353 TI - Fasting and nonfasting LDL cholesterol: to measure or calculate? PMID- 19282354 TI - Acidification of blood is superior to sodium fluoride alone as an inhibitor of glycolysis. AB - BACKGROUND: Sodium fluoride is the preferred agent to inhibit glycolysis. Its action is not immediate, however, and complete inhibition is delayed for up to 4 hours. A more effective method is needed. Acidification of blood combined with the addition of NaF and EDTA appears to be such a method. We studied whether acidification was indeed more effective than NaF. METHODS: We conducted 6 independent studies over a 10-month period at 3 Quest Diagnostics laboratory sites. In each study, we drew venous blood from 6-24 nonfasting employee volunteers into 3 or 4 different serum- or plasma-collection tubes, which were stored under different conditions and aliquoted at different times. We analyzed the aliquots in duplicate by means of a hexokinase-based enzymatic method. RESULTS: The mean glucose concentration decreased by 0.3% at 2 h and by 1.2% at 24 h when blood was drawn into tubes containing citrate buffer, NaF, and EDTA. In contrast, the mean glucose concentration decreased by 4.6% at 2 h and by 7.0% at 24 h when blood was drawn into tubes containing NaF and sodium oxalate. CONCLUSIONS: Acidification should replace NaF alone as the recommended method for obtaining an accurate glucose concentration. Diagnostic cut points based on blood samples collected into tubes containing NaF as the only inhibitor of glycolysis are likely to be too low. PMID- 19282355 TI - Familial dysalbuminemic hyperthyroxinemia: a persistent diagnostic challenge. PMID- 19282356 TI - Day 5 morning serum cortisol predicts hypothalamic-pituitary-adrenal function after transsphenoidal surgery for pituitary tumors. AB - BACKGROUND: Adrenal insufficiency is a complication of transsphenoidal surgery (TSS) for pituitary adenoma, and correct identification of patients requiring glucocorticoid replacement is important. Controversy exists over which early postoperative 9 AM cortisol concentration reliably predicts hypothalamic pituitary-adrenal (HPA) axis reserve, as defined by the insulin tolerance test (ITT). METHODS: Data were reviewed for 36 patients undergoing TSS followed by day 5 postoperative 9 AM cortisol measurement and ITT 6 weeks postsurgery. All patients received postoperative glucocorticoid replacement, which was discontinued if the 9 AM serum cortisol was >300 nmol/L. RESULTS: Of 23 patients who failed the ITT (peak cortisol <500 nmol/L), 20 also had a day 5, 9 AM serum cortisol <300 nmol/L. Nine of 13 patients who passed the ITT had a day 5, 9 AM cortisol >300 nmol/L. The cutoff cortisol concentration of 300 nmol/L had 86.9% (66.4%-97.2%) diagnostic sensitivity, 69.2% (38.6%-90.9%) diagnostic specificity, and 83.3% (61.8%-94.5%) positive predictive value (PPV) for detecting secondary adrenal insufficiency. Increasing the cutoff to 392 nmol/L resulted in 100% (85.2%-100%) sensitivity, 46.1% (19.2%-74.9%) specificity, and 76.6% (57.3% 89.4%) PPV. Decreasing the cutoff to 111 nmol/L resulted in 100% (75.3%-100%) specificity and 100% (67.9%-100%) PPV, although sensitivity was 47.8% (26.8% 69.4%). CONCLUSIONS: A day 5 post-TSS 9 AM serum cortisol <111 nmol/L reliably detects secondary adrenal insufficiency, and concentrations >392 nmol/L support intact HPA function. Because concentrations of 111-392 nmol/L are poorly predictive of HPA function, glucocorticoid replacement should continue in such cases until definitive testing is performed using an ITT. PMID- 19282357 TI - Dealing with missing predictor values when applying clinical prediction models. AB - BACKGROUND: Prediction models combine patient characteristics and test results to predict the presence of a disease or the occurrence of an event in the future. In the event that test results (predictor) are unavailable, a strategy is needed to help users applying a prediction model to deal with such missing values. We evaluated 6 strategies to deal with missing values. METHODS: We developed and validated (in 1295 and 532 primary care patients, respectively) a prediction model to predict the risk of deep venous thrombosis. In an application set (259 patients), we mimicked 3 situations in which (1) an important predictor (D-dimer test), (2) a weaker predictor (difference in calf circumference), and (3) both predictors simultaneously were missing. The 6 strategies to deal with missing values were (1) ignoring the predictor, (2) overall mean imputation, (3) subgroup mean imputation, (4) multiple imputation, (5) applying a submodel including only the observed predictors as derived from the development set, or (6) the "one-step sweep" method. We compared the model's discriminative ability (expressed by the ROC area) with the true ROC area (no missing values) and the model's estimated calibration slope and intercept with the ideal values of 1 and 0, respectively. RESULTS: Ignoring the predictor led to the worst and multiple imputation to the best discrimination. Multiple imputation led to calibration intercepts closest to the true value. The effect of the strategies on the slope differed between the 3 scenarios. CONCLUSIONS: Multiple imputation is preferred if a predictor value is missing. PMID- 19282358 TI - S-Nitrosothiols in blood: does photosensitivity explain a 4-order-of-magnitude concentration range? PMID- 19282359 TI - Tumor cell targeting using folate-conjugated fluorescent quantum dots and receptor-mediated endocytosis. AB - BACKGROUND: Luminescent nanobioprobes with cell-targeting specificity are likely to find important applications in bioanalysis, biomedicine, and clinical diagnosis. Quantum dots (QDs) are unique and promising materials for such a purpose because of their fluorescence and large surface area for attaching cell targeting molecules. METHODS: We produced water-dispersible QDs by coating hydrophobic QDs with small amphiphilic polyethylene glycol (PEG) molecules via hydrophobic interactions. We covalently coupled folate (FA) onto the water dispersible PEG-coated QDs (PEG-QDs) to produce FA-coupled PEG-QDs (FA-PEG-QDs). RESULTS: These FA-PEG-QD nanoparticles functioned as fluorescent nanobioprobes that specifically recognized folate receptors (FRs) overexpressed in human nasopharyngeal cells (KB cells) but not in an FR-deficient lung carcinoma cell line (A549 cells). Using confocal fluorescence microscopy, we demonstrated uptake of FA-PEG-QDs by KB cells but no uptake of folate-free PEG-QDs. The specificity of this receptor-mediated internalization was confirmed by comparing the uptake by KB vs A549 cells. CONCLUSIONS: Our results suggest that such cell-targeting fluorescent nanobioprobes are potentially very powerful tools for recognizing target cells and delivering and tracking drugs and other therapeutic materials. PMID- 19282360 TI - Serum-based test of the pathologic breakdown of type I collagen fibers. PMID- 19282361 TI - Rapid and long-term adaptations in gait symmetry following unilateral step training in people with hemiparesis. AB - BACKGROUND AND OBJECTIVE: Evidence for specific physical interventions that improve walking symmetry in individuals with hemiparesis poststroke is limited. The aim of this study was to investigate the rapid and prolonged effects of unilateral step training (UST) on step length asymmetry (SLA) in people with hemiparesis. SUBJECTS AND DESIGN: Eighteen individuals with chronic hemiparesis and substantial SLA during overground walking participated in a single-group, pretest-posttest study. The study consisted of 2 phases, with 10 subjects participating in each phase; 2 subjects participated in both phases. INTERVENTIONS AND MEASUREMENTS: To investigate rapid effects of UST, the participants completed a 20-minute session of UST on a treadmill with their unimpaired limb, with the impaired limb held stationary off the treadmill. Data for spatiotemporal gait parameters during overground walking at self-selected and fastest speeds were collected prior to and following UST, with follow-up measurements at 1 day and 1 week. To investigate the prolonged effects, the participants completed ten 20-minute sessions of UST. Data for spatiotemporal gait parameters were collected prior to training as well as after every third session, with follow-up measurements at 1 and 2 weeks. RESULTS: Immediately following UST, SLA tested during fast-paced overground walking improved by up to 13% (49% reduced to a 36% SLA), with changes retained for up to 24 hours. Following 10 sessions of UST, SLA improved significantly, with changes retained for up to 2 weeks. LIMITATIONS: Despite repeated baseline measurements, the absence of a control group was a limitation. Furthermore, stepping characteristics during UST were not quantified. CONCLUSION: Unilateral step training may improve spatiotemporal patterns in people with substantial gait asymmetry poststroke. Repeated training may be necessary for maintenance of adaptations. PMID- 19282362 TI - Short-term efficacy of upper-extremity exercise training in patients with chronic airway obstruction: a systematic review. AB - BACKGROUND, OBJECTIVES, AND MEASUREMENTS: Patients with chronic airway obstruction (CAO) frequently experience dyspnea and fatigue during activities performed by accessory muscles of ventilation, which competitively participate in arm elevation. This systematic review of randomized controlled trials (RCTs) concerning patients with CAO addresses the effects of upper-extremity exercise training (UEET), added to lower-extremity training or comprehensive pulmonary rehabilitation, on the following patient-centered outcomes: exercise capacity, symptoms, ability to perform daily activities, and health-related quality of life. METHODS: Studies were retrieved using comprehensive database and hand search strategies. Two independent reviewers determined study eligibility based on inclusion criteria. A detailed description of treatments was mandatory. Reviewers rated study quality and extracted information on study methods, design, intervention, and results. RESULTS: Forty publications were evaluated. Four RCTs met the inclusion criteria but had serious methodological limitations, which introduce possible biases that reduce their internal validity. The outcomes measured were heterogeneous, and the results were inconsistent regarding maximal exercise capacity, dyspnea, and health-related quality of life. No effect of UEET was demonstrated for measures of arm fatigue. LIMITATIONS AND CONCLUSIONS: The limited methodological quality of the studies retrieved prevented us from performing a meta-analysis, the results of which could be misleading. This systematic review shows that there is limited evidence examining UEET and that the evidence available is of poor quality. Therefore, a recommendation for the inclusion or exclusion of UEET in pulmonary rehabilitation programs for individuals with CAO is not possible. Further research is needed to definitively ascertain the effects of this training modality on patient-centered outcomes. PMID- 19282364 TI - Minireview: fetal-maternal hormonal signaling in pregnancy and labor. AB - Mechanisms underlying the initiation of parturition remain unclear. Throughout most of pregnancy, uterine quiescence is maintained by elevated progesterone acting through progesterone receptor (PR). Although in most mammals, parturition is associated with a marked decline in maternal progesterone, in humans, circulating progesterone and uterine PR remain elevated throughout pregnancy, suggesting a critical role for functional PR inactivation in the initiation of labor. Both term and preterm labor in humans and rodents are associated with an inflammatory response. In preterm labor, intraamniotic infection likely provides the stimulus for increased amniotic fluid interleukins and migration of inflammatory cells into the uterus and cervix. However, at term, the stimulus for this inflammatory response is unknown. Increasing evidence suggests that the developing fetus may produce physical and hormonal signals that stimulate macrophage migration to the uterus, with release of cytokines and activation of inflammatory transcription factors, such as nuclear factor kappaB (NF-kappaB) and activator protein 1 (AP-1), which also is activated by myometrial stretch. We postulate that the increased inflammatory response and NF-kappaB activation promote uterine contractility via 1) direct activation of contractile genes (e.g. COX-2, oxytocin receptor, and connexin 43) and 2) impairment of the capacity of PR to mediate uterine quiescence. PR function near term may be compromised by direct interaction with NF-kappaB, altered expression of PR coregulators, increased metabolism of progesterone within the cervix and myometrium, and increased expression of inhibitory PR isoforms. Alternatively, we propose that uterine quiescence during pregnancy is regulated, in part, by PR antagonism of the inflammatory response. PMID- 19282365 TI - The PPARgamma2 A/B-domain plays a gene-specific role in transactivation and cofactor recruitment. AB - We have previously shown that adenoviral expression of peroxisome proliferator activated receptors (PPARs) leads to rapid establishment of transcriptionally active complexes and activation of target gene expression within 5-8 h after transduction. Here we have used the adenoviral delivery system combined with expression array analysis to identify novel putative PPARgamma target genes in murine fibroblasts and to determine the role of the A/B-domain in PPARgamma mediated transactivation of genomic target genes. Of the 257 genes found to be induced by PPARgamma2 expression, only 25 displayed A/B-domain dependency, i.e. significantly reduced induction in the cells expressing the truncated PPARgamma lacking the A/B-domain (PPARgammaCDE). Nine of the 25 A/B-domain-dependent genes were involved in lipid storage, and in line with this, triglyceride accumulation was considerably decreased in the cells expressing PPARgammaCDE compared with cells expressing full-length PPARgamma2. Using chromatin immunoprecipitation, we demonstrate that PPARgamma binding to genomic target sites and recruitment of the mediator component TRAP220/MED1/PBP/DRIP205 is not affected by the deletion of the A/B-domain. By contrast, the PPARgamma-mediated cAMP response element-binding protein (CREB)-binding protein (CBP) and p300 recruitment to A/B-domain-dependent target genes is compromised by deletion of the A/B-domain. These results indicate that the A/B-domain of PPARgamma2 is specifically involved in the recruitment or stabilization of CBP- and p300-containing cofactor complexes to a subset of target genes. PMID- 19282366 TI - Genetic interactions of the androgen and Wnt/beta-catenin pathways for the masculinization of external genitalia. AB - In most mammals, the sexually dimorphic development of embryos is typically achieved by the differentiation of the external genitalia. Hence, the sexual distinction of mammalian newborns is based on the external genital structure. Although it was shown in the 1940s and 1950s that androgen from the testes establishes the male sexual characteristics, the involvement of nongonadal and locally produced masculine effectors remains totally unknown. It is noteworthy that the disorders of fetal masculinization, including hypospadias, one of the most frequent birth defects, occur at a high frequency. Furthermore, their causative factors remain unclear. In this study, the involvement of the coordinated actions of androgen and the growth factor systems was genetically analyzed for the first time on mammalian reproductive organ formation. The results demonstrated that the Wnt/beta-catenin pathway is indispensable masculine factor for the external genital development. The bilateral mesenchymal region adjacent to the urethral plate epithelium displayed a sexually dimorphic activity of Wnt/beta-catenin signaling. Loss- and gain-of-function beta-catenin mutants displayed altered sexual development of the external genitalia. These results indicate the novel functions of the Wnt/beta-catenin pathway as a locally expressed masculine effector. This could be the first genetic study analyzing the roles of the genetic interactions between androgen and locally expressed growth factor signaling during the development of reproductive organs. These results also shed new insight on the reproductive genetics and the causative factors of genital disorders. PMID- 19282367 TI - Oncogenic ras blocks the cAMP pathway and dedifferentiates thyroid cells via an impairment of pax8 transcriptional activity. AB - A deranged differentiation is often a landmark of transformed cells. We used a thyroid cell line expressing an inducible Ras oncoprotein in order to study the hierarchy of molecular events leading to suppression of thyroid-specific gene expression. We find that, upon Ras activation, there is an immediate global down regulation of thyroid differentiation, which is associated with an inhibition of the cAMP signaling pathway. We demonstrate that an unusual negative cross talk between Ras oncogene and the cAMP pathway induces inactivation of the transcription factor Pax8 that we propose as a crucial event in Ras-induced dedifferentiation. PMID- 19282368 TI - High milk consumers have an increased risk of folate receptor blocking autoantibody production but this does not affect folate status in Spanish men and women. AB - Folate receptor (FR)-blocking autoantibodies (FR-autoantibodies) have been reported in women with neural tube defect-affected pregnancies and subfertility and in children with progressive neurodevelopment disorders. We investigated their prevalence and association with folate status and milk intake in adults unexposed to folic acid fortification. A cross-sectional study of a randomly selected representative sample of a Spanish population (aged 18-75 y) stratified by age and gender was performed. Plasma and red cell folate, plasma cobalamin, fasting plasma total homocysteine (tHcy) concentration, methylenetetrahydrofolate reductase C677T polymorphism, and FR-autoantibody titer were determined in blood samples from 787 fasting participants. Lifestyle data were collected and milk intake estimated from a 3-d dietary record. FR-autoantibody prevalence was 7.2% [0.30 +/- 0.27 nmol (mean +/- SD) FR blocked/L], equally affecting men and women of all ages. Plasma and red cell folate and tHcy did not differ between carriers and noncarriers of FR-autoantibodies. Milk intake was higher in carriers (225 +/- 199 g/d) than in noncarriers (199 +/- 147 g/d) (P < 0.01). The risk of having FR autoantibodies increased progressively with increasing quintile of milk intake and was significant in the highest quintile (> or =307 g/d) compared with the lowest (< or =67 g/d) [odds ratio (OR), 2.41 [95% CI: 1.02, 5.69]; P < 0.05; linear trend, P = 0.02]. We concluded that FR-autoantibodies occur in men and women of all ages and do not affect indicators of folate status such as plasma and red cell folate and tHcy. Higher milk intake is associated with increased risk of having FR-autoantibodies. PMID- 19282369 TI - Intestinal nitrogen recycling and utilization in health and disease. AB - The role of intestinal microflora in digestive and metabolic processes has received increasing attention from researchers and clinicians. Both enterocytes and small intestine luminal microorganisms can degrade peptides and amino acids (AA). Further, enterocytes can utilize ammonia via glutamate, glutamine, citrulline, and urea synthesis, whereas luminal microbes will deaminate AA, hydrolyze luminal urea, and recycle this ammonia by synthesis of new microbial cells. Although, undoubtedly, some indispensable AA may arise from N cycling and microbial synthesis in the intestinal lumen, the actual net impact on protein nutrition status appears to be limited in humans and animals. Moreover, potential contributions of the recycled N as colonic luminal microbial proteins to AA in blood depend on colonic protein digestion and AA absorption. Finally, new evidence indicates that gut microbial metabolism may be enhanced by prebiotics and probiotics, with the prospects of new treatment paradigms for eliminating undesirable secondary N metabolites and ameliorating complications in whole-body N metabolism under the conditions of intestinal stress, liver disease, and kidney failure. PMID- 19282370 TI - Glutamate is the major anaplerotic substrate in the tricarboxylic acid cycle of isolated rumen epithelial and duodenal mucosal cells from beef cattle. AB - In this study, we aimed to determine the contribution of substrates to tricarboxylic acid (TCA) cycle fluxes in rumen epithelial cells (REC) and duodenal mucosal cells (DMC) isolated from Angus bulls (n = 6) fed either a 75% forage (HF) or 75% concentrate (HC) diet. In separate incubations, [(13)C(6)]glucose, [(13)C(5)]glutamate, [(13)C(5)]glutamine, [(13)C(6)]leucine, or [(13)C(5)]valine were added in increasing concentrations to basal media containing SCFA and a complete mixture of amino acids. Lactate, pyruvate, and TCA cycle intermediates were analyzed by GC-MS followed by (13)C-mass isotopomer distribution analysis. Glucose metabolism accounted for 10-19% of lactate flux in REC from HF-fed bulls compared with 27-39% in REC from HC and in DMC from bulls fed both diets (P < 0.05). For both cell types, as concentration increased, an increasing proportion (3-63%) of alpha-ketoglutarate flux derived from glutamate, whereas glutamine contributed <3% (P < 0.05). Although leucine and valine were catabolized to their respective keto-acids, these were not further metabolized to TCA cycle intermediates. Glucose, glutamine, leucine, and valine catabolism by ruminant gastrointestinal tract cells has been previously demonstrated, but in this study, their catabolism via the TCA cycle was limited. Further, although glutamate's contribution to TCA cycle fluxes was considerable, it was apparent that other substrates available in the media also contributed to the maintenance of TCA fluxes. Lastly, the results suggest that diet composition alters glucose, glutamate, and leucine catabolism by the TCA cycle of REC and DMC. PMID- 19282371 TI - Milk-fat depression and lipid repartitioning in lactating dairy cows. PMID- 19282372 TI - Visualization of plastids in pollen grains: involvement of FtsZ1 in pollen plastid division. AB - Visualizing organelles in living cells is a powerful method to analyze their intrinsic mechanisms. Easy observation of chlorophyll facilitates the study of the underlying mechanisms in chloroplasts, but not in other plastid types. Here, we constructed a transgenic plant enabling visualization of plastids in pollen grains. Combination of a plastid-targeted fluorescent protein with a pollen specific promoter allowed us to observe the precise number, size and morphology of plastids in pollen grains of the wild type and the ftsZ1 mutant, whose responsible gene plays a central role in chloroplast division. The transgenic material presented in this work is useful for studying the division mechanism of pollen plastids. PMID- 19282373 TI - Potential errors in electron transport rates calculated from chlorophyll fluorescence as revealed by a multilayer leaf model. AB - Increasingly, photosynthetic electron transport rate is being calculated from chlorophyll fluorescence measurements. The fluorescence signal is a complex mixture of contributions from different depths within the mesophyll. One condition required for electron transport calculated from fluorescence to represent the rate accurately is that the ratio of photosynthetic capacity to light absorbed be constant throughout the leaf. In order to explore the fluorescence properties of leaves where this assumption is not true, a new approximation for phiPSII is used to generate F'm and Fs values throughout the leaf. Fs is assumed to be proportional to the amount of light absorbed from the fluorescence measuring beam and constant, i.e. independent of the actinic irradiance or CO2 concentration. This assumption is validated by measurements from Eucalyptus maculata, Flaveria bidentis and Triticum aestivum, with two different types of fluorometer, where irradiance or CO2 response curves were measured with normal or inverted leaf orientations. The new approach enables fluorescence values to be generated at each layer in a multilayer model. Two applications using this approach are presented. First, the model is used to show that when quantum yield varies through a leaf, then fluorescence will lead to an incorrect estimate of electron transport rate. Secondly, since chlorophyll fluorescence is also used to calculate the CO2 concentration at the sites of carboxylation within chloroplasts, Cc, the model is also used to show that Cc may vary with depth. Significant variation in Cc through the mesophyll could lead to an apparent dependence of internal conductance on irradiance or CO2. PMID- 19282374 TI - A randomized controlled trial of massage therapy in children with sickle cell disease. AB - OBJECTIVE: This randomized controlled trial investigated the short-term effects of massage therapy on youth with SCD and their parents. METHODS: Thirty-four children and adolescents, and their parents were assigned to a massage therapy or an attention control group. Parents were trained in massage in their homes once a week for 4 weeks, with instructions to provide nightly massages. Families in the control group were visited weekly by a research assistant. Participants completed measures of depression and anxiety, functional status, pain intensity, medication use, and service utilization. RESULTS: Parents in the massage therapy group reported higher levels of depression and anxiety following the intervention. Youth in this group showed higher levels of functional status, and lower levels of depression, anxiety, and pain. Health service utilization rates were unchanged from pre- to post-intervention. CONCLUSIONS: These results offer preliminary support for parent-delivered massage therapy as an intervention for SCD pain. PMID- 19282375 TI - Nelfinavir/ritonavir reduces acinar injury but not inflammation during mouse caerulein pancreatitis. AB - There is no clinical treatment that reduces acinar injury during pancreatitis. Human immunodeficiency virus (HIV) protease inhibitors (PI), including nelfinavir (NFV) and ritonavir (RTV), may reduce the rate of pancreatitis in HIV-infected patients. Since permeability transition pore (PTPC)-mediated mitochondrial dysfunction occurs during pancreatitis, and we have shown that PI prevents PTPC opening, we studied its effects in a model of pancreatitis. The effect of NFV plus RTV (NFV/RTV) or vehicle on caerulein-induced pancreatitis in mice was compared by measuring changes in mitochondrial membrane potential in vitro and cytochrome c leakage in vivo. Histological and inflammatory makers were also compared. NFV/RTV improved DiOC6 retention in acini exposed to caerulein in vitro. In vivo NFV prevented cytosolic leakage of cytochrome c and reduced pancreatic acinar injury, active caspase-3 staining, TUNEL-positive acinar cells, and serum amylase (P < 0.05). Conversely, trypsin activity, serum cytokine levels, and pancreatic and lung inflammation were unaffected. NFV/RTV reduces pancreatic injury and acinar cell death in experimental mouse caerulein-induced pancreatitis but does not impact inflammation. PMID- 19282376 TI - Functional role of J domain of cysteine string protein in Ca2+-dependent secretion from acinar cells. AB - The heat shock protein 70 family members Hsc70 and Hsp70 are known to play a protective role against the onset of experimental pancreatitis, yet their molecular function in acini is unclear. Cysteine string protein (CSP-alpha) is a zymogen granule (ZG) membrane protein characterized by an NH(2)-terminal "J domain" and a central palmitoylated string of cysteine residues. The J domain functions as a cochaperone by modulating the activity of Hsc70/Hsp70 family members. A role for CSP-alpha in regulating digestive enzyme exocytosis from pancreas was investigated by introducing CSP-alpha truncations into isolated acini following their permeabilization with Perfringolysin O. Incubation of acini with CSP-alpha(1-82), containing the J domain, significantly augmented Ca(2+) stimulated amylase secretion. Effects of CSP-alpha(1-82) were concentration dependent, with a maximum 80% increase occurring at 200 microg/ml of protein. Although CSP-alpha(1-82) had no effects on basal secretion measured in the presence of < or =10 nM free Ca(2+), it did significantly augment GTP-gammaS induced secretion under basal Ca(2+) conditions by approximately 25%. Mutation of the J domain to abolish its cochaperone activity failed to augment Ca(2+) stimulated secretion, implicating the CSP-alpha/Hsc70 cochaperone system as a regulatory component of the secretory pathway. CSP-alpha physically associates with vesicle-associated membrane protein 8 (VAMP 8) on ZGs, and the CSP-alpha VAMP 8 interaction was dependent on amino acids 83-112 of CSP-alpha. Immunofluorescence analysis of acinar lobules or purified ZGs confirmed the CSP alpha colocalization with VAMP 8. These data establish a role for CSP-alpha in regulating digestive enzyme secretion and suggest that CSP-alpha and Hsc70 modulate specific soluble N-ethylmaleimide-sensitive attachment receptor interactions necessary for exocytosis. PMID- 19282377 TI - Substance P promotes expansion of human mesenteric preadipocytes through proliferative and antiapoptotic pathways. AB - White adipose tissue is intimately involved in the regulation of immunity and inflammation. We reported that human mesenteric preadipocytes express the substance P (SP)-mediated neurokinin-1 receptor (NK-1R), which signals proinflammatory responses. Here we tested the hypothesis that SP promotes proliferation and survival of human mesenteric preadipocytes and investigated responsible mechanism(s). Preadipocytes were isolated from mesenteric fat biopsies during gastric bypass surgery. Proliferative and antiapoptotic responses were delineated in 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4 sulfophenyl)-2H-tetrazolium (MTS), bromodeoxyuridine (BrdU), caspase-3, and TUNEL assays, as well as Western immunoanalysis. SP (10(-7) M) increased MTS and proliferation (BrdU) and time dependently (15-30 min) induced Akt, EGF receptor, IGF receptor, integrin alphaVbeta3, phosphatidylinositol 3-kinase, and PKC-theta phosphorylation. Furthermore, pharmacological antagonism of Akt and PKC-theta activation significantly attenuated SP-induced preadipocyte proliferation. Exposure of preadipocytes to the proapoptotic Fas ligand (FasL, 100 microM) resulted in nuclear DNA fragmentation (TUNEL assay), as well as increased cleaved poly (ADP-ribose) polymerase, cleaved caspase-7, and caspase-3 expression. Cotreatment with SP almost completely abolished these responses in a NK-1R dependent fashion. SP (10(-7) M) also time dependently stimulated expression 4E binding protein 1 and phosphorylation of p70 S6 kinase, which increased protein translation efficiency. SP increases preadipocyte viability, reduces apoptosis, and stimulates proliferation, possibly via cell cycle upregulation and increased protein translation efficiency. SP-induced proliferative and antiapoptotic pathways in fat depots may contribute to development of the creeping fat and inflammation characteristic of Crohn's disease. PMID- 19282378 TI - Alpha-melanocyte-stimulating hormone counteracts the suppressive effect of UVB on Nrf2 and Nrf-dependent gene expression in human skin. AB - Human skin is constantly exposed to UV light, the most ubiquitous environmental stressor. Here, we investigated the expression and regulation of Nrf1-3, transcription factors crucially involved in protection against oxidative stress in human skin cells in vitro, ex vivo, and in situ. In particular, we examined whether alpha-MSH, a UV-induced peptide, is capable of modulating Nrf2 and Nrf dependent gene expression. Nrf1, -2, and -3 were found to be expressed in various cutaneous cell types in vitro. Surprisingly, UVB irradiation at physiological doses (10 mJ/cm(2)) reduced Nrf2 and Nrf-dependent gene expression in normal keratinocytes and melanocytes in vitro as well as ex vivo in skin organ cultures. alpha-MSH alone significantly increased Nrf2 as well as Nrf-dependent heme oxygenase-1, gamma-glutamylcysteine-synthetase, and glutathione-S-transferase Pi gene expression in both keratinocytes and melanocytes. This effect of alpha-MSH occurred at physiological doses and was due to transcriptional induction, mimicked by the artificial cAMP inducer forskolin, and blocked by protein kinase A pathway inhibition. In silico promoter analysis of Nrf2 further identified several putative binding sites for activator protein 1 and cAMP response element binding protein, transcription factors typically activated by alpha-MSH. Importantly, alpha-MSH prevented or even overcompensated the UVB-induced suppression of Nrf2 and Nrf-dependent genes not only in normal keratinocytes and melanocytes in vitro but also in skin organ cultures. These findings, for the first time, show regulation of Nrf2 and Nrf-dependent genes by alpha-MSH. Our data also highlight a novel facet in the cytoprotective and antioxidative effector mechanisms of alpha-MSH and perhaps of related melanocortin peptides. PMID- 19282379 TI - Ovariectomy and 17beta-estradiol replacement do not alter beta-amyloid levels in sheep brain. AB - The benefits of estrogen replacement as a preventative treatment for Alzheimer's disease (AD) are subject to debate. Because the effects of estrogen depletion and replacement on accumulation of the neurotoxic beta-amyloid (A beta) peptide in transgenic animal models of AD have been variable, we examined A beta levels and oxidative stress in a nontransgenic animal model. Sheep have traditionally been used as a model for human reproduction; however because they share 100% sequence homology with the human form of A beta, they may also have potential as a nontransgenic model for A beta biology. The effect of ovariectomy and estrogen replacement administered for 6 months via slow-release implant was examined in the brain of 4.5-yr-old sheep. A beta levels were measured by ELISA, and protein levels of the amyloid precursor protein (APP), APP C-terminal fragments (C100), and presenilin-1 were examined semiquantitatively by Western blot as markers of APP processing. Markers of oxidative stress were examined semiquantitatively by Western blot [4-hydroxy-2(E)-nonenal] and oxyblot (protein carbonyls). We found no effects of estrogen depletion and supplementation in terms of AD-related biochemical markers, including A beta levels, APP processing, and oxidative stress levels. Evidence of a trend toward increased P450 side-chain cleavage enzyme levels in the hippocampus of ovariectomized and estrogen supplemented sheep suggests that neurosteroidogenesis may compensate for gonadal estrogen depletion; however, these findings cannot explain the lack of effect of estrogen supplementation on APP processing. It is possible that supraphysiological doses of estrogen are necessary to yield antiamyloidogenic and antioxidative benefits in ovariectomized sheep. PMID- 19282380 TI - The rhythmic secretion of mating-induced prolactin secretion is controlled by prolactin acting centrally. AB - Artificial copulomimetic cervical stimulation (CS) induces an immediate release of oxytocin (OT) and prolactin (PRL) followed by a daily PRL rhythm characterized by nocturnal and diurnal surges. Although we have shown that the initial release of PRL is induced by the immediate release of OT, we tested whether the PRL that is released in response to CS is responsible for the initiation and maintenance of the subsequent PRL surges. Thus, we injected OVX rats centrally or peripherally with ovine PRL (oPRL) at 2200 h. Central oPRL induced PRL surges in OVX rats that were similar in size and timing to those of CS rats, whereas peripheral oPRL induced surges that were of smaller amplitude and delayed. We then infused a PRL antagonist (S179D, 0.1 ng/h) centrally into OVX and OVX-CS rats and measured the release of endogenous PRL and the activity of neuroendocrine dopaminergic neurons. Central infusion of S179D did not influence basal PRL secretion in OVX rats but prevented the expression of the CS-induced PRL surges and the accompanying noontime increase of CS-induced dopaminergic activity when continued for 3 d. However, central infusion of S179D only on the day of CS did not prevent the daily rhythm of PRL surges. These results demonstrate that PRL acts centrally to induce the PRL rhythm and that PRL in the brain is essential for the maintenance but not for the initiation of the CS induced rhythmic PRL surges. PMID- 19282381 TI - Systemic administration of C-type natriuretic peptide as a novel therapeutic strategy for skeletal dysplasias. AB - Skeletal dysplasias are a group of genetic disorders characterized by severe impairment of bone growth. Various forms of them add to produce a significant morbidity and mortality, yet no efficient drug therapy has been developed to date. We previously demonstrated that C-type natriuretic peptide (CNP), a member of the natriuretic peptide family, is a potent stimulator of endochondral bone growth. Furthermore, we exhibited that targeted overexpression of a CNP transgene in the growth plate rescued the impaired bone growth observed in a mouse model of achondroplasia (Ach), the most frequent form of human skeletal dysplasias, leading us to propose that CNP may prove to be an effective treatment for this disorder. In the present study, to elucidate whether or not the systemic administration of CNP is a novel drug therapy for skeletal dysplasias, we have investigated the effects of plasma CNP on impaired bone growth in Ach mice that specifically overexpress CNP in the liver under the control of human serum amyloid P component promoter or in those treated with a continuous CNP infusion system. Our results demonstrated that increased plasma CNP from the liver or by iv administration of synthetic CNP-22 rescued the impaired bone growth phenotype of Ach mice without significant adverse effects. These results indicate that treatment with systemic CNP is a potential therapeutic strategy for skeletal dysplasias, including Ach, in humans. PMID- 19282382 TI - Sexual differentiation of the spinal nucleus of the bulbocavernosus is not mediated solely by androgen receptors in muscle fibers. AB - The spinal nucleus of the bulbocavernosus (SNB) neuromuscular system is a highly conserved and well-studied model of sexual differentiation of the vertebrate nervous system. Sexual differentiation of the SNB is currently thought to be mediated by the direct action of perinatal testosterone on androgen receptors (ARs) in the bulbocavernosus/levator ani muscles, with concomitant motoneuron rescue. This model has been proposed based on surgical and pharmacological manipulations of developing rats as well as from evidence that male rats with the testicular feminization mutation (Tfm), which is a loss of function AR mutation, have a feminine SNB phenotype. We examined whether genetically replacing AR in muscle fibers is sufficient to rescue the SNB phenotype of Tfm rats. Transgenic rats in which wild-type (WT) human AR is driven by a human skeletal actin promoter (HSA-AR) were crossed with Tfm rats. Resulting male HSA-AR/Tfm rats express WT AR exclusively in muscle and nonfunctional Tfm AR in other tissues. We then examined motoneuron and muscle morphology of the SNB neuromuscular system of WT and Tfm rats with and without the HSA-AR transgene. We observed feminine levator ani muscle size and SNB motoneuron number and size in Tfm males with or without the HSA-AR transgene. These results indicate that AR expression in skeletal muscle fibers is not sufficient to rescue the male phenotype of the SNB neuromuscular system and further suggest that AR in other cell types plays a critical role in sexual differentiation of this system. PMID- 19282383 TI - The alternative noncoding exons 1 of aromatase (Cyp19) gene modulate gene expression in a posttranscriptional manner. AB - Aromatase (Cyp19) is a key enzyme in estrogen biosynthesis and an important target in endocrine therapy for estrogen receptor (ER)-positive postmenopausal breast cancer. Aromatase transcription is driven by multiple tissue-specific promoters, which result in the production of various mRNA transcripts that contain an alternative noncoding exon 1 followed by a common protein-coding region. Transcriptional activity of these promoters is the only known determinant for aromatase protein abundance in a given tissue or cellular context. To determine whether aromatase expression could be influenced by additional regulatory mechanisms, we used a common heterologous promoter to drive the expression of multiple aromatase cDNA sequences that differ only by the alternative exon 1 sequence. These expression vectors gave rise to vastly different levels of aromatase mRNA and protein in multiple cell lines examined. Furthermore, the relative abundance of several mRNA variants did not correlate with that of the corresponding protein product. The variation in mRNA and protein levels is most likely due to a negative effect of certain alternative exons 1 on RNA stability and protein translation. Deletional analyses indicate that the 5' regions of the adipose tissue-specific exons I.3 and I.4 contain the cis-acting elements responsible for modulation of aromatase levels. Thus, our work uncovers an important role of the alternative exons 1 in posttranscriptional regulation of aromatase gene expression. PMID- 19282384 TI - Mechanisms of protein kinase C signaling in the modulation of 3',5'-cyclic adenosine monophosphate-mediated steroidogenesis in mouse gonadal cells. AB - The protein kinase C (PKC) signaling pathway plays integral roles in the expression of the steroidogenic acute regulatory (StAR) protein that regulates steroid biosynthesis in steroidogenic cells. PKC can modulate the activity of cAMP/protein kinase A signaling involved in steroidogenesis; however, its mechanism remains obscure. In the present study, we demonstrate that activation of the PKC pathway, by phorbol 12-myristate 13-acetate (PMA), was capable of potentiating dibutyryl cAMP [(Bu)(2)cAMP]-stimulated StAR expression, StAR phosphorylation, and progesterone synthesis in both mouse Leydig (MA-10) and granulosa (KK-1) tumor cells. The steroidogenic potential of PMA and (Bu)(2)cAMP was linked with phosphorylation of ERK 1/2; however, inhibition of the latter demonstrated varying effects on steroidogenesis. Transcriptional activation of the StAR gene by PMA and (Bu)(2)cAMP was influenced by several factors, its up regulation being dependent on phosphorylation of the cAMP response element binding protein (CREB). An oligonucleotide probe containing a CREB/activating transcription factor binding region in the StAR promoter was found to bind nuclear proteins in PMA and (Bu)(2)cAMP-treated MA-10 and KK-1 cells. Chromatin immunoprecipitation studies revealed that the induction of phosphorylated CREB was tightly correlated with in vivo protein-DNA interactions and recruitment of CREB binding protein to the StAR promoter. Ectopic expression of CREB binding protein enhanced CREB-mediated transcription of the StAR gene, an event that was markedly repressed by the adenovirus E1A oncoprotein. Further studies demonstrated that the activation of StAR expression and steroid synthesis by PMA and (Bu)(2)cAMP was associated with expression of the nuclear receptor Nur77, indicating its essential role in hormone-regulated steroidogenesis. Collectively, these findings provide insight into the mechanisms by which PKC modulates cAMP/protein kinase A responsiveness involved in regulating the steroidogenic response in mouse gonadal cells. PMID- 19282385 TI - Insulin-like growth factor and epidermal growth factor treatment: new approaches to protecting steatotic livers against ischemia-reperfusion injury. AB - Hepatic steatosis is a major risk factor in ischemia-reperfusion (I/R). IGF binding proteins (IGFBPs) modulate IGF-I action by transporting circulating IGF-I to its sites of action. Epidermal growth factor (EGF) stimulates IGF-I synthesis in vitro. We examined the effect of IGF-I and EGF treatment, separately or in combination, on the vulnerability of steatotic livers to I/R. Our results indicated that I/R impaired IGF-I synthesis only in steatotic livers. Only when a high dose of IGF-I (400 microg/kg) was given to obese animals did they show high circulating IGF-I:IGFBP levels, increased hepatic IGF-I levels, and protection against damage. In lean animals, a dose of 100 microg/kg IGF-I protected nonsteatotic livers. Our results indicated that the combined administration of IGF-I and EGF resulted in hepatic injury parameters in both liver types similar to that obtained by IGF-I and EGF separately. IGF-I increased egf expression in both liver types. The beneficial role of EGF on hepatic I/R injury may be attributable to p38 inhibition in nonsteatotic livers and to PPAR gamma overexpression in steatotic livers. In conclusion, IGF-I and EGF may constitute new pharmacological strategies to reduce the inherent susceptibility of steatotic livers to I/R injury. PMID- 19282386 TI - Expression of a gonadotropin-releasing hormone receptor-simian virus 40 T-antigen transgene has sex-specific effects on the reproductive axis. AB - The GnRH receptor (GnRHR) responds to pulsatile GnRH signals to coordinate pituitary gonadotropin synthesis and secretion. Previously, a 1.2-kb fragment of the 5'-flanking region isolated from the mouse GnRHR gene was shown to target expression to pituitary gonadotropes in vivo. The 1.2-kb gene promoter fused to the simian virus 40 large T antigen (TAg) was used to generate transgenic mice that form gonadotrope-derived pituitary tumors at 4-5 months of age. Transgenic female mice have hypogonadotropic hypogonadism, infantile gonads, and are infertile throughout their life span, whereas males remain reproductively intact until their tumors become large. We hypothesized that the targeted TAg expression causes a sex-specific disruption of the reproductive axis at the level of the pituitary gland. To test this hypothesis, we characterized the pituitary gonadotropin beta-subunit and TAg expression patterns, and measured plasma gonadotropin and gonadal steroid levels in female and male mice before and after pituitary tumor development. TAg expression was observed in transgenic females and males 15 d of age, before tumor development. Interestingly, and in contrast to the transgenic males, pituitary LH beta and FSH beta subunit protein levels, and plasma LH and FSH levels, were reduced in transgenic females. Reproductive organs in transgenic female mice remained underdeveloped but were normal in transgenic males. We conclude that the expression of the TAg transgene driven by the GnRHR gene promoter results in female-specific infertility due to disruption of gonadotropin production and secretion even before tumor development. PMID- 19282387 TI - A novel androgen receptor amino terminal region reveals two classes of amino/carboxyl interaction-deficient variants with divergent capacity to activate responsive sites in chromatin. AB - The androgen receptor (AR) is an important signaling molecule in multiple tissues, yet its mode of action and cell-specific activities remain enigmatic. AR function has been best studied in the prostate, in which it is essential for growth and homeostasis of the normal organ as well as each stage of cancer development. Investigation of mechanisms responsible for continued AR action that evolve during prostate cancer progression or after hormonal management of the disease have been instructive in defining AR signaling pathways. In the current paper, we use sequence similarity and the collocation of somatic mutations in prostate cancer to define residues 501-535 of the AR amino-terminal domain as an important mediator of receptor function. Specifically, the 501-535 region is required for optimal interaction of the amino-terminal domain with both the p160 coactivator, nuclear receptor coactivator-2, and the AR-ligand binding domain in the amino/carboxyl (N/C) interaction. The N/C interaction is decreased by deletion of the 501-535 region but is distinct from deletion of the (23)FQNLF(27) peptide in that it does not affect the capacity of the AR to activate transcription from a chromatin integrated reporter or recruitment of the receptor to androgen-responsive loci in vivo. Collectively, we have been able to outline two classes of N/C-deficient AR variant that are divergent in their capacity to act in a chromatin context, thereby further defining the interplay between N/C interaction and coregulator recruitment via multiple receptor domains. These mechanisms are likely to be key determinants of the cell and promoter specific activities of the AR. PMID- 19282388 TI - Interference of a mutant thyroid hormone receptor alpha1 with hepatic glucose metabolism. AB - Mice expressing the mutant thyroid hormone receptor TRalpha1R384C, which has a 10 fold reduced affinity to the ligand T(3), exhibit hypermetabolism due to an overactivation of the sympathetic nervous system. To define the consequences in the liver, we analyzed hepatic metabolism and the regulation of liver genes in the mutant mice. Our results showed that hepatic phosphoenolpyruvate carboxykinase was up-regulated and pyruvate kinase mRNA down-regulated, contrary to what observed after T(3) treatment. In contrast, mice expressing a mutant TRalpha1L400R specifically in the liver did not show a dysregulation of these genes; however, when the TRalpha1L400R was expressed ubiquitously, the hepatic phenotype differed from TRalpha1R384C animals, suggesting that the localization of the mutation plays an important role for its consequences on glucose metabolism. Furthermore, we observed that glycogen stores were completely depleted in TRalpha1R384C animals, despite increased gluconeogenesis and decreased glycolysis. Exposure of the mutant mice to high maternal levels of thyroid hormone during fetal development leads to a normal liver phenotype in the adult. Our results show how genetic and maternal factors interact to determine the metabolic setpoint of the offspring and indicate an important role for maternal thyroid hormone in the susceptibility to metabolic disorders in adulthood. PMID- 19282389 TI - Estrogens regulate posttranslational modification of neural cell adhesion molecule during the estrogen-induced gonadotropin surge. AB - Estrogen-induced synaptic plasticity (EISP) in the periventricular area (PVA) of the hypothalamus is necessary for the preovulatory gonadotropin surge. Because in situ enzymatic desialization of hypothalamic polysialylated (PSA) neural cell adhesion molecule (NCAM) blocked EISP, we examined the presence and amount of NCAM isotopes, PSA-NCAM, and sialylation enzymes in microdissected mouse hypothalamus tissues from proestrous afternoon [peak of estrogens and nadir of arcuate nucleus (AN) synapses] and metestrous morning (nadir of estrogens and highest AN synapses). Immunohistochemistry confirmed immunoreactive (ir) PSA-NCAM staining in the perineural spaces of the PVA. The extent of staining was cycle dependent, with more dense and complete profiles of individual neurons limned by the ir-PSA-NCAM staining on proestrus and less on metestrus. Western blots showed that high levels of ir-PSA-NCAM on proestrus are accompanied by diminished ir NCAM-140 and -180 but not ir-NCAM-120 and the reverse on metestrus (P < 0.05). To evaluate the increase of sialylated NCAM at the expense of desialylated protein, expression of the responsible polysialyltransferase enzymes polysialyltransferase (ST8Sia IV) and sialyltransferase (ST8Sia II) mRNA levels were measured using RT PCR. Both polysialyltransferase and sialyltransferase mRNA are more abundant on proestrus than metestrus (P < 0.05), indicating that these enzymes are regulated by estrogens. These results support estrogen-regulated formation and extrusion of hydrophilic PSA-NCAM into perineural spaces in the PVA as part of the mechanism of EISP. PMID- 19282390 TI - Agreement between task-based estimates of the full-shift noise exposure and the full-shift noise dosimetry. AB - Noise assessments have been conducted using full-shift dosimetry and short-term task-based measurements. Advantages of the task-based method include the opportunity to directly identify high-noise exposure tasks and to target control measures, as well as obtain estimates of task-based full-shift exposures; however, there is little empirical evidence comparing the two methods. National Institute for Occupational Safety and Health assessed noise exposures at three industrial facilities using dosimetry and task-based methods with the objective of comparing the two strategies and assessing the degree of agreement and causes of disagreement. Eight indices of task-based full-shift exposures were created from task-based sampling using three methods to assess time-at-task (direct observation by industrial hygienist, end-of-shift worker estimates and supervisor estimates) and three methods to assign noise levels to tasks [direct measurement, arithmetic mean (AM) and geometric mean (GM)]. We assessed aspects of agreement (precision, bias and absolute agreement) using Bland-Altman plots and concordance correlation coefficient (CCC). Overall, the task-based methods worked fairly well, with mean biases less than +/-2.8 dBA and precision ranges of 3.3-4.4 dBA. By all measures, task-based full-shift estimates based on supervisor assessment of time-at-task agreed most poorly with the dosimetry data. The task-based full shift estimates based on worker estimates of time-at-task generally agreed as well as those based on direct observation. For task noise level, task-based full shift estimates based on directly measured task agreed the best with dosimetry data, while agreement for task-based indices based on task AM or GM was variable. Overall, the task-based full-shift estimates based on direct observation task and direct measured task noise level achieved the best agreement with the dosimetry data (CCC 0.84) with 95% of their differences being within 7.4 dBA and 56% of the differences <3 dBA. For this index, a high degree of accuracy was observed (accuracy coefficient = 0.96) with major cause of disagreement arising from a lack of precision (precision coefficient = 0.88). When the measurements were classified by job characteristics, significant improvements in the degree of agreement were observed in the low job mobility, low job complexity and low job variability categories. Our data suggest that a high degree of absolute agreement can be achieved between the task-based and dosimetry-based estimates of full shift exposures. The task-based approach that uses worker reports combined with task AM or GM levels is similar to the more time-intensive direct observation method to estimate full-shift exposures. PMID- 19282391 TI - The CMP-legionaminic acid pathway in Campylobacter: biosynthesis involving novel GDP-linked precursors. AB - The sialic acid-like sugar 5,7-diacetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto nonulosonic acid, or legion-aminic acid, is found as a virulence-associated cell surface glycoconjugate in the Gram-negative bacteria Legionella pneumophila and Campylobacter coli. L. pneumophila serogroup 1 strains, causative agents of Legionnaire's disease, contain an alpha2,4-linked homopolymer of legionaminic acid within their lipopolysaccharide O-chains, whereas the gastrointestinal pathogen C. coli modifies its flagellin with this monosaccharide via O-linkage. In this work, we have purified and biochemically characterized 11 candidate biosynthetic enzymes from Campylobacter jejuni, thereby fully reconstituting the biosynthesis of legionaminic acid and its CMP-activated form, starting from fructose-6-P. This pathway involves unique GDP-linked intermediates, likely providing a cellular mechanism for differentiating between this and similar UDP linked pathways, such as UDP-2,4-diacetamido-bacillosamine biosynthesis involved in N-linked protein glycosylation. Importantly, these findings provide a facile method for efficient large-scale synthesis of legionaminic acid, and since legionaminic acid and sialic acid share the same D-glycero-D-galacto absolute configuration, this sugar may now be evaluated for its potential as a sialic acid mimic. PMID- 19282393 TI - Methadone isomers and their role in pain management. PMID- 19282394 TI - Novobiocin is a potent inhibitor for human organic anion transporters. AB - Organic anion transporters (OATs) mediate the body disposition of a diverse array of environmental toxins and clinically important drugs. Previous studies have shown that novobiocin, an inhibitor for breast cancer resistance proteins (BCRP), inhibited organic anion transport. However, its interactions with specific OATs are unknown. In the current study, we characterized the inhibitory effects of novobiocin on the function of human OATs (hOAT)1, hOAT3, and hOAT4. Kinetic study revealed that novobiocin inhibited OAT-mediated uptake in a competitive manner, with K(i) of 14.87 +/- 0.40 microM for hOAT1, K(i) of 4.77 +/- 1.12 microM for hOAT3, and K(i) of 90.50 +/- 7.50 microM for hOAT4. Furthermore, the cis- and trans-inhibition feature of novobiocin demonstrated that novobiocin was a potent inhibitor but not a substrate for hOAT1 (IC(50) = 34.76 +/- 0.31 microM), hOAT3 (IC(50) = 4.987 +/- 0.35 microM), and hOAT4 (IC(50) = 92.68 +/- 0.34 microM). We further showed that the effects of novobiocin on OATs were not mediated through a change in transporter protein abundance on the plasma membrane. Taken together, we conclude that novobiocin seems to interact with the substrate-binding sites of OATs from both the intracellular and the extracellular sides, and this interaction interferes with the substrate-binding site(s) on respective carriers, leading to an apparent reduction in carriers available for the substrates. Because BCRP is often expressed in the same tissue where multiple OATs are identified such as liver, kidney and placenta, when dissecting the contribution of BCRP to drug disposition using novobiocin as an inhibitor, its inhibitory effect to OATs has to be taken into consideration. PMID- 19282395 TI - Characterization of dasatinib and its structural analogs as CYP3A4 mechanism based inactivators and the proposed bioactivation pathways. AB - Dasatinib was approved in 2006 for the treatment of imatinib-resistant chronic myelogenous leukemia and functions primarily through the inhibition of BCR-ABL and Src kinase. Dasatinib is extensively metabolized in humans by CYP3A4. In this study, we report that the bioactivation of dasatinib by CYP3A4 proceeds through a reactive intermediate that leads to CYP3A4 inactivation with K(I) = 6.3 microM and k(inact) = 0.034 min(-1). The major mechanism of inactivation proceeds through hydroxylation at the para-position of the 2-chloro-6-methylphenyl ring followed by further oxidation, forming a reactive quinone-imine, similar to the reactive intermediates formed by acetaminophen and diclofenac. Formation of a reactive imine-methide was also detected but appears to be a minor pathway. When glutathione was added to human liver microsomal incubations, dasatinib glutathione adducts were detected. Numerous dasatinib analogs were synthesized in an effort to understand what modifications would block the formation of reactive intermediates during dasatinib metabolism. It is interesting to note that blocking the site of hydroxylation with a methyl group was not effective because a reactive imine-methide was formed, nor was blocking the site with fluorine because the fluorine was removed through an oxidative defluorination mechanism and the reactive quinone-imine was still formed. Numerous analogs are presented that did effectively block the formation of glutathione adducts and prevent the inactivation of CYP3A4. PMID- 19282396 TI - Impact of gut microbiota on intestinal and hepatic levels of phase 2 xenobiotic metabolizing enzymes in the rat. AB - Using immunoblotting, we compared levels of phase 2 enzymes in liver, small intestine, cecum, and colon of germ-free and control rats (reassociated with rat intestinal microbiota). In addition, colonic levels were studied after association with human intestinal microbiota. The glutathione transferases (GSTs) studied, gastrointestinal glutathione peroxidase (GPX2), both epoxide hydrolases (EPHXs), and N-acetyltransferase (NAT) 1, were detected in all tissues. GPX2 and GSTP1 were highest in large bowel; the other enzymes of this group were highest in liver. NAT2 was found in the large bowel but not in the liver or small bowel. Sulfotransferases (SULTs) were detected in liver but were absent in small intestine; two forms were present at moderate levels in the large intestine. Strong gender-dependent differences were observed for several enzymes in liver but not in gut. Colonic levels in germ-free animals differed from those in control animals (* indicates statistical significance) for GSTA1/2 (4.0*- and 5.0*-fold in males and females, respectively), GSTA4 (1.5*/1.9*-fold), GSTM1 (1.1/1.5*-fold), EPHX1 (3.5*/2.4*-fold), EPHX2 (1.4/2.1*-fold), SULT1B1 (0.4*/0.6*-fold), SULT1C2 (1.3/1.6*-fold), and NAT2 (1.4/1.5*-fold). Smaller effects were observed when rats were colonized with human, compared with rat, intestinal bacteria. Cecal enzyme levels in germ-free rats were changed similarly to those in colon. No effects were seen in small intestine. In liver, SULT1A1, SULT1C1, and SULT1C2 were elevated in germ-free animals of both genders (1.5- to 2.6-fold); hepatic EPHX2 was elevated 1.6-fold in females. In conclusion, intestinal microbiota can affect levels of xenobiotic-metabolizing enzymes in large intestine and liver, but the effects observed were moderate compared with tissue-dependent expression differences. PMID- 19282397 TI - Quantitative prediction of in vivo profiles of CYP3A4 induction in humans from in vitro results with a reporter gene assay. AB - Although primary human hepatocytes are commonly used for induction studies, the evaluation method is associated with several problems. More recently, a reporter gene assay has been suggested to be an alternative, although the contribution of only transfected nuclear receptors can be evaluated. The aim of the present study was to establish a method by which the extent of in vivo CYP3A4 induction in humans can be quantitatively predicted based on in vitro results with a reporter gene assay. From previous reports, we calculated in vivo induction ratios (R(in vivo)) caused by prototypical inducers based on the alterations in the hepatic intrinsic clearance of probe drugs. Next, we derived equations by which these R(in vivo) values can be predicted from the results of a reporter gene assay. To use the data obtained from a reporter gene assay, rifampicin was used as a reference drug. The correction coefficient (CC), which is used to quantitatively correlate the activity of inducers between in vitro and in vivo situations, was calculated by comparing the predicted data with the observed R(in vivo) values for rifampicin. With the calculated CC value, good correlations were found between the predicted and observed R(in vivo) values for other inducers such as phenobarbital, phenytoin, and omeprazole. Taken together, with the equations derived in the present study, we have been able to predict the extent of in vivo induction of human CYP3A4 by inducers in a time-dependent and quantitative manner from in vitro data. PMID- 19282398 TI - Long-lasting inhibition of the transporter-mediated hepatic uptake of sulfobromophthalein by cyclosporin a in rats. AB - Cyclosporin A (CsA) is a well known inhibitor of the organic anion-transporting polypeptide (OATP/Oatp) family transporters, causing a large number of transporter-mediated drug-drug interactions in clinical situations. In the present study, we examined the inhibitory effect of CsA on the hepatic uptake of sulfobromophthalein (BSP) in rats, focusing on a long-lasting inhibition. Twenty one hours after the subcutaneous administration of CsA, the hepatic clearance of BSP was decreased. The liver uptake index study revealed that hepatic uptake of BSP was reduced in CsA-treated rats for at least 3 days. Comparison of uptake studies using isolated hepatocytes prepared from control and CsA-treated rats showed that hepatic uptake in CsA-treated rats was decreased. In primary cultured hepatocytes, after preincubation with CsA, the uptake of [(3)H]BSP was reduced even after removal of CsA from the incubation buffer although a preincubation time dependence was not observed. However, the expression of Oatp1a1 and Oatp1b2, which are involved in the hepatic uptake of BSP, and the amount of intrahepatic glutathione, a driving force of Oatp1a1, did not change in CsA-treated rats. Thus, we can conclude that CsA modulates the transporter function sustainably. It can cause a potent in vivo drug-drug interaction. The modulation of transporters is not caused by reduced expression or driving force of transporters. It may be affected by CsA accumulated in the liver or its metabolites. The inhibitory effect of CsA on the transporter-mediated uptake of BSP cannot be explained by a simple competitive mechanism and a novel mechanism should be considered. PMID- 19282399 TI - The Gigogne flap: an original technique for an optimal pulp reconstruction. AB - The reconstruction of digital pulp defects is still a challenge in hand surgery. Several reconstructive techniques are available, but in some cases, their advancement capacities are not sufficient for the flap to cover the whole defect. The Gigogne flap is a new, very simple and safe technique. It consists of harvesting two cutaneous flaps successively on the same neurovascular digital palmar bundle. The first plasty may be chosen among the well-known flaps usually applied in pulp reconstruction. The second plasty, the Gigogne flap, is an advancement VY plasty done on the main cutaneous paddle of the first proximal flap. We performed 15 Gigogne flaps in 13 patients, who suffered zone II and III digital pulp amputations. The Gigogne flap increased the advancement capacity of the proximal flap as well as restoring the physiological pulp fatty pad, thus reconstructing a functional and aesthetic fingertip. PMID- 19282400 TI - Effect of triangular ligament tears on distal radioulnar joint instability and evaluation of three clinical tests: a biomechanical study. AB - Using 11 fresh/frozen cadaver specimens, instability of the distal radioulnar joint (DRUJ) during passive wrist motion was measured by motion analysis and three clinical tests (ulno-carpal stress test, piano-key test, and DRUJ ballottement test) were performed. Release of the triangular ligament from the palmar approach was performed to simulate instability of the DRUJ. Displacement of the distal radius against the ulnar head was measured at various degrees of forearm rotation. Release of the triangular ligament at the ulnar fovea and around the base of the ulnar styloid process caused various degrees of DRUJ instability during passive wrist motion. After complete triangular ligament release, only the DRUJ ballottement test demonstrated a statistically significant degree of accuracy in the evaluation of distal radioulnar joint instability. PMID- 19282401 TI - Perforation of the third extensor compartment by the drill bit during palmar plating of the distal radius. AB - The frequency of penetration into the third extensor compartment when drilling the distal screw holes was assessed for four different palmar distal radius plates in 160 cadaver forearms. Penetration into this compartment occurred in 43%. Different plates had different penetration rates: 3.5 LCP four-hole locking T-plate: 20%, 3.5 LCP three-hole locking plate: 42.5%, 2.4 palmar LCP standard plate and the 2.4 palmar LCP buttress plate: 55%. When using a palmar plate on the distal radius, the surgeon risks penetrating into the third extensor compartment. PMID- 19282402 TI - Radial tunnel syndrome: emphasis on the superficial branch of the radial nerve. AB - Reported success rates for decompressing the radial nerve in patients with radial tunnel syndrome vary between 10 and 95%. The combined treatment, releasing both the posterior interosseous nerve and the superficial branch of the radial nerve, has been described only three times, but seems to show more consistent success rates compared with releasing the posterior interosseous nerve alone. We present the results of decompressing the superficial branch of the radial nerve only, the anatomical basis for this approach and a description of the surgical technique. Our results are comparable to the results of the combined treatment. Eleven of 12 patients were satisfied with the results of the operation. This study indicates that pain in patients with radial tunnel syndrome may be treated successfully by surgical decompression of the superficial branch of the radial nerve. PMID- 19282403 TI - The blood supply of the scaphoid bone. AB - Scaphoid vascularisation was investigated using macroscopic and microscopic techniques in 12 uninjured, formalin fixed cadaver hands. A good blood supply of the scaphoid bone from palmar, dorsal and radial vessel groups with a variety of anastomoses was found which should provide sufficient collateral blood flow from adjacent regions in some patients. Since blood supply is available from the palmar circulation, a dorsal approach to the scaphoid bone is possible. PMID- 19282404 TI - Congenital duplication of the palm syndrome: gene analysis and the molecular basis of its clinical features. AB - Congenital duplication of the palm is a rare syndrome with the following features: the dorsal aspects of both hands have thick palmar skin with no hair or nails; bilateral ulnar ray deficiency; short hypoplastic upper limbs; and severe lower limb abnormalities. In this paper, we report a new case of congenital duplication of the palm syndrome, provide its gene analysis identifying the responsible gene mutation in exon 4 of the WNT7a gene, and detail the molecular basis of its clinical features. PMID- 19282405 TI - Outcome following coronal fractures of the hamate. AB - This is a case series of 14 hamate body fractures, in the coronal plane, with 12 patients having an associated metacarpal injury. Treatment was either conservative or operative and was decided according to the degree of fracture displacement and presence of carpometacarpal subluxation. Patient follow-up was 14.8 months on average, using DASH scores and postoperative radiographs to determine functional and anatomical outcomes. Undisplaced fractures were treated in a cast with good outcome. Postoperative radiographs of patients treated with Kirschner wires for displaced fractures showed incomplete reduction and the outcomes were less favourable. Rigid internal fixation gave excellent radiographic results, although these did not correlate with functional outcome in all patients. PMID- 19282406 TI - Flexor tendon repair in zone 2 using a six-strand 'figure of eight' suture. AB - The tensile strength of three different flexor tendon repair techniques were tested in vitro: the modified Kessler technique (a two-strand repair), two 'figure of eight' sutures (a four-strand repair) and three 'figure of eight' sutures (a six-strand repair). The mean breaking forces for the three techniques were 48.0 N, 73.1 N and 93.3 N, respectively, and the differences were highly significant (p < 0.0001). In a prospective clinical study, a total of 45 patients (50 fingers) with clean-cut complete lacerations of both flexor tendons in zone 2 were included. The protocol used the three 'figure of eight' suture techniques for (profundus only) tendon repair, 'venting' of the pulleys, and post-operative immediate active range of motion that ensured full active extension of the proximal interphalangeal joint. One repair (2%) ruptured. In the remaining 49 repairs, the result was considered excellent in 39 (78%) and good in 10 (20%) using the Strickland and Glogovac grading system. PMID- 19282407 TI - A midterm follow-up study of bioreconstructive polylactide scaffold implants in metacarpophalangeal joint arthroplasty in rheumatoid arthritis patients. AB - This paper presents the results of a prospective study of 80 metacarpophalangeal joint arthroplasties, in which biodegradable polylactide 96/4 copolymer scaffolds were used. Twenty-three rheumatoid arthritis patients were assessed at an average of 59 months after operation, which exceeds the resorption time of P(L/D)LA 96/4 according to animal experiments. Palmar subluxation exceeded half of the bone thickness in 39 joints before operation and in nine at the last follow-up. Ulnar deviation decreased from 25 degrees to 5 degrees , extension deficit from 32 degrees to 15 degrees and active flexion from 76 degrees to 63 degrees . The results are comparable with published data on silicone implant arthroplasties. Implant resorption did not induce any significant osteolysis in the medium term and the restoration of the structure and function of the hand was maintained after implant resorption, probably as the guided fibrous tissues had replaced the dissolved implant. PMID- 19282408 TI - Keratoderma hereditarium mutilans (Vohwinkel syndrome). AB - Keratoderma hereditarium mutilans, or Vohwinkel syndrome, is a very rare genetic skin condition which causes palmoplantar hyperkeratosis and constricting rings of the fingers and toes. Approximately 50 cases have been reported in the literature with only three having been managed surgically. All three had a high rate of recurrence and unfavourable results in the long term. We report two more cases managed surgically with a follow up of 5 and 8 years respectively. Our experience suggests that the use of full thickness grafts to line the released contractures does not work in the long term as the grafts become raised and painful, requiring multiple revisions. Surgical correction was easy to achieve but difficult to maintain and achieved poor outcomes in general. We therefore feel that the indication for surgical treatment should be a neurovascular compromise. PMID- 19282409 TI - Hammering versus drilling of sharp and obtuse trocar-point k-wires. AB - Kirschner wire characteristics affect the heating of bone during insertion and the subsequent strength of fixation. We inserted 90 sharp and 90 obtuse trocar tip K-wires into 90 fresh frozen human cadaver metacarpals using either a drill or a pneumatic hammer. The temperature elevation, insertion time and extraction force were measured for four K-wire insertion combinations: drilling sharp; drilling obtuse; hammering sharp; hammering obtuse. Hammering resulted in significantly lower temperature elevations than drilling. Hammering sharp K-wires resulted in the highest extraction forces. The first and fifth metacarpals showed significantly lower temperature elevations than the other metacarpals, while the insertion time was significantly higher in the second and third metacarpal than in the other metacarpals. Hammering sharp trocar-tip K-wires minimises thermal damage to bone and gives the strongest fixation. PMID- 19282410 TI - Pre- and postoperative Guyon's canal pressure change in endoscopic carpal tunnel release: correlation with transient postoperative Guyon's canal syndrome. AB - Perioperative Guyon's canal and carpal canal pressure in one-forearm portal endoscopic carpal tunnel release surgery were measured in resting position and during active power gripping in 66 hands. This was done using the continuous infusion technique with a local anaesthetic and without pneumatic tourniquet. Immediate mean postoperative Guyon's canal and carpal canal pressure decreased in both measurements. During active power gripping, postoperative Guyon's canal pressure was less than 40 mmHg in 61 hands, however, this increased to over 40 mmHg in five hands. In these five hands, Guyon's canal syndrome did not develop. Guyon's canal and carpal canal pressures were only correlated during postoperative active power gripping. It remains unclear whether immediate postoperative Guyon's canal pressure correlates with higher pressures a few days later as reported in cases of transient postoperative Guyon's canal syndrome. PMID- 19282412 TI - Influence of core suture geometry on tendon deformation and gap formation in porcine flexor tendons. AB - The effects of core suture geometry on the mechanics of failure in flexor tendon surgery are investigated. Forty porcine flexor tendons were repaired using a Kessler; a Kessler-Pennington; a double Kessler; a continuous Kessler; and a cruciate repair. At maximum breaking strength, the cruciate repair gapped more then the double Kessler (12.8 mm vs 9.1 mm), but the double Kessler was less strong (37N vs 45 N). Transverse narrowing was 22% and 24% for the Kessler and the Kessler-Pennington, 11% for the double Kessler, and 0% for the continuous Kessler and the cruciate repair. Kessler-type sutures failed by suture breakage and the cruciate repair by pull-out. Under load, the transverse part of the Kessler sutures narrows, allowing longitudinal parts to lengthen, leading to gapping. The double Kessler shortened transverse segment decreases gapping. Eliminating a transverse component (the cruciate repair) decreased gapping, but the cruciate failed at higher loads by suture pull-out. PMID- 19282413 TI - Conservative treatment of the cubital tunnel syndrome. AB - Conservative treatment of the cubital tunnel syndrome was evaluated in a randomised study of 70 patients with mild or moderate symptoms (Dellon, 1989). All patients were informed about the cause of symptoms and allocated to three groups: night splinting, nerve gliding and control. Evaluation consisted of Canadian Occupational Performance Measure, visual analogue pain scales, strength measurements and neurophysiological examination, before treatment and after six months. Fifty-seven patients were followed for six months. Fifty-one (89.5%) were improved at the follow-up. There were no significant differences between the groups in any of the recorded variables. Night splints and nerve gliding exercises did not add favourably. Routine neurophysiological examination seems unnecessary since 76% of the patients with typical symptoms had normal findings and 75% with pathological findings improved. Patients with mild or moderate symptoms have a good prognosis if they are informed of the causes of the condition and how to avoid provocation. PMID- 19282414 TI - Anatomical and radiological assessment of trapezial osteotomy for trapezial dysplasia in early trapeziometacarpal joint arthritis. AB - Opening wedge osteotomy of the trapezial saddle was reported first by Kapandji and Heim (2002) as a possible surgical treatment for early trapeziometacarpal joint arthritis. This study evaluates the feasibility and anatomical risks of the procedure. Ten upper limbs from fresh cadavers were used for this anatomical and radiological study. A dorsolateral opening wedge osteotomy with a 10 degrees correction was performed on each specimen. CT scans and AP radiographs of all the wrists were performed before and after osteotomy to assess the correction of the slope angle, defined as the angle between the longitudinal axis of the second metacarpal and the axis of the trapeziometacarpal articulation. On standard radiographs, mean trapezial slope was 126 degrees preoperatively and 117 degrees postoperatively. On sagittal and coronal CT reconstructions, one case of trapezial translation and two impingements between trapezoid and trapezium bones were identified. Trapezial osteotomy is technically demanding with several possible complications, but merits further study for young patients with Eaton stage 1 or 2 osteoarthritis and an abnormal trapezial slope. PMID- 19282415 TI - Spontaneous intracerebral hemorrhage in a patient with multiple sclerosis and tumefactive demyelinating lesion. AB - In this report, we discuss the occurrence of intracerebral hemorrhage in a patient with multiple sclerosis during treatment with natalizumab, a humanized monoclonal antibody against the alpha4beta1-integrin. Hemorrhage was located in a previously tumefactive demyelinating lesion. The mechanisms of leukocyte recruitment to the sites of inflammation through interaction of leukocyte alpha4beta1-integrins and endothelial vascular cell adhesion molecule-1 are well known. However, alpha4beta1-integrins are also expressed on endothelial cells and CD34(+) bone marrow-derived progenitor cells, controlling several key pathways in angiogenesis. Neovascularization may contribute to tissue repair, particularly in large inflammatory cerebral lesions with increased vascular fragility. We discuss possible interaction of natalizumab with angiogenesis during tissue repair. PMID- 19282416 TI - Abnormalities in brain synchronization are correlated with cognitive impairment in multiple sclerosis. AB - BACKGROUND: Cognitive functions are supported by brain networks and they are highly dependent on the integrity of long white matter tracts that mediate the information flow between such distant cortical areas. Brain damage in multiple sclerosis (MS) may produce cognitive impairment by preferentially damaging these tracts, thereby impairing brain synchrony. Auditory amplitude modulation following responses (AMFR), are oscillatory steady-state responses to rhythmic auditory stimuli that indirectly measure brain synchrony. OBJECTIVE: To study the effect of MS lesions in brain synchrony and its relationship with cognitive function. METHODS: We assessed the correlation between cognitive performance, as assessed with the brief repeatable battery-neuropsychology (BRB-N), and the AMFR in a group of 27 MS patients and 22 healthy controls. RESULTS: The maximal AMFR frequency - but not the amplitude - in the 30-60 Hz range was lower in patients with cognitive impairment than in patients with no cognitive impairment or the healthy controls (39.79 Hz, 43.85 Hz, and 43.84 Hz, respectively, P < 0.05). Indeed, the frequency of the AMFR was negatively correlated with the scores obtained in verbal memory, attention, and executive function. The multiple regression analysis indicates that the AMFR was the best predictor of the BRB-N scores after controlling for potential confounding factors such as age, education, disability, and years of disease evolution. CONCLUSIONS: These results suggest that the loss of synchronization in different central nervous system (CNS) pathways caused by demyelinating lesions might involve both the slowing of brain oscillatory activity and less efficient cognitive processing. PMID- 19282417 TI - A randomized trial to investigate the effects of functional electrical stimulation and therapeutic exercise on walking performance for people with multiple sclerosis. AB - BACKGROUND: Functional electrical stimulation (FES), is a means of producing a contraction in a paralyzed or weak muscle to enable function through electrical excitation of the innervating nerve. OBJECTIVE: This two-group randomized trial assessed the effects of single channel common peroneal nerve stimulation on objective aspects of gait relative to exercise therapy for people with secondary progressive multiple sclerosis (SPMS). METHODS: Forty-four people with a diagnosis of SPMS and unilateral dropped foot completed the trial. Twenty patients were randomly allocated to a group receiving FES and the remaining 24 to a group receiving a physiotherapy home exercise program for a period of 18 weeks. RESULTS: The exercise group showed a statistically significant increase in 10 m walking speed and distance walked in 3 min, relative to the FES group who showed no significant change in walking performance without stimulation. At each stage of the trial, the FES group performed to a significantly higher level with FES than without for the same outcome measures. CONCLUSION: Exercise may provide a greater training effect on walking speed and endurance than FES for people with SPMS. FES may provide an orthotic benefit when outcome is measured using the same parameters. More research is required to investigate the combined therapeutic effects of FES and exercise for this patient group. PMID- 19282418 TI - Modification of vincristine dosing during concomitant azole therapy in adult acute lymphoblastic leukemia patients. AB - OBJECTIVE: Vincristine is an important component in the treatment of acute lymphoblastic leukemia (ALL) and is now the backbone of therapy in the induction and consolidation phases of this disease. Proper dosing of vincristine is required to maximize disease control while avoiding toxicity. The gastrointestinal toxicity of vincristine such as decreased peristalsis can potentially be increased if the CYP 3A4 enzyme is inhibited. This interaction may become more prevalent with increasing use of CYP 3A4 inhibitors such as the azole antifungals. Since azoles are increasingly being used for prophylaxis and treatment of fungal infections in this patient population, an assessment of vincristine dosing and toxicity is the first step to constructing guidelines for the coadministration of these agents. METHODS: ALL patients !18 years of age receiving vincristine-based therapy from August 2003 through December 2007 with or without azole therapy were included. Data was collected using electronic patient medical records and the pharmacy system (RxTFC). Information was entered into a database for this retrospective study. Patients were separated into two arms; vincristine with azoles and vincristine only. Patient demographic information, chemotherapy regimen, vincristine-induced symptoms, and concurrent strong CYP 3A4 inhibitors and inducers were collected. RESULTS: A total of 50 patients received vincristine of which 29 (58%) had concurrent azole therapy. No patients received concurrent major CYP 3A4 inhibitors and the baseline characteristics were similar between groups. Vincristine dosing modifications were more common in the azole group (58.6 vs. 23.8%; p = 0.02). The mean dose reduction of vincristine when combined with an azole was 46.5%. Symptoms of decreased peristalsis were more common in patients receiving azoles (65.5 vs. 28.6%; p = 0.019) and on average occurred after the second vincristine dose. Symptoms occurred in 50, 75, and 66.6% of patients receiving fluconazole, voriconazole, and posaconazole, respectively. Patients were more likely to have an incomplete course of vincristine when receiving azole therapy (48.3 vs. 9.5%; p = 0.004). CONCLUSION: Caution should be used with the coadministration of vincristine and azoles. It is recommended that institutional guidelines be developed to standardize care for patients receiving vincristine with azole therapy. Potential measures to avoid this interaction include revisiting azole prophylaxis in this patient group and being judicious in azole selection. PMID- 19282419 TI - Effects of antipsychotics and vitamin C on the formation of reactive oxygen species. AB - There is evidence that reactive oxygen species (ROS) are involved in the pathophysiology of psychiatric disorders such as schizophrenia. Indirect biochemical alterations of ROS formation have been shown for patients treated with antipsychotics as well as for untreated patients. Only one study measured directly the ROS formation after treatment with antipsychotics by using electron spin resonance spectroscopy. The aim of the present examination was to demonstrate the effects of haloperidol, clozapine and olanzapine in concentrations of 18, 90 and 180 MUg/mL on the formation of ROS in the whole blood of rats by using electron spin resonance spectroscopy after incubation for 30 min. To test the protective capacity of vitamin C we incubated the highest concentration of each drug with vitamin C (1 mM). Under all treatment conditions, olanzapine led to a significantly higher formation of ROS compared with control conditions, whereas in the cases of haloperidol and clozapine the two higher concentrations induced a significantly enhanced formation of ROS. Vitamin C reduced the ROS production of all drugs tested and for haloperidol and clozapine the level of significance was reached. Our study demonstrated that antipsychotics induce the formation of ROS in the whole blood of rats, which can be reduced by the application of vitamin C. PMID- 19282421 TI - Agmatine disrupts prepulse inhibition of acoustic startle reflex in rats. AB - Agmatine is a guanidine-amine formed by the enzymatic decarboxylation of arginine. Agmatine has been proposed to be a neuromodulator and its downstream derivatives, the polyamines, have been suggested to be responsible for sensory gating deficits seen in schizophrenia. In this study, male Wistar rats underwent treatments with agmatine, vehicle or other agents known to alter sensory gating in an experimental paradigm of prepulse inhibition (PPI) of the acoustic startle response. Apomorphine (1 mg/kg s.c.), a nonselective dopamine agonist known to disrupt PPI responses, was injected as the positive reference. Neither apomorphine nor agmatine (40-160 mg/kg i.p.) induced effects on the intensity of startle reflex without a prepulse. However, apomorphine or agmatine (160 mg/kg i.p.) disrupted the PPI of acoustic startle reflex. Furthermore, when given 30 min prior, agmatine acted additively with apomorphine's effect on PPI. In an attempt to gain more insight, haloperidol (1 and 2 mg/kg i.p.), clozapine (2.5 7.5 mg/kg i.p.) or quetiapine (2.5 and 7.5 mg/kg i.p.) was also injected prior to agmatine (160 mg/kg i.p.). Haloperidol (1 mg/kg) and clozapine (2.5 and 5 mg/kg) were able to prevent the PPI-disrupting effects of apomorphine. However, none of these antipsychotics prevent the PPI-disrupting effects of agmatine. These results suggest that agmatine disrupts the PPI of acoustic startle reflex of rats in a fundamentally different manner than apomorphine does. It may also have a critical role in the pathogenesis of sensorimotor gating-related dysfunctions. PMID- 19282420 TI - Conditioned place preference and locomotor activity in response to methylphenidate, amphetamine and cocaine in mice lacking dopamine D4 receptors. AB - Methylphenidate (MP) and amphetamine (AMPH) are the most frequently prescribed medications for the treatment of attention-deficit/hyperactivity disorder (ADHD). Both drugs are believed to derive their therapeutic benefit by virtue of their dopamine (DA)-enhancing effects, yet an explanation for the observation that some patients with ADHD respond well to one medication but not to the other remains elusive. The dopaminergic effects of MP and AMPH are also thought to underlie their reinforcing properties and ultimately their abuse. Polymorphisms in the human gene that codes for the DA D4 receptor (D4R) have been repeatedly associated with ADHD and may correlate with the therapeutic as well as the reinforcing effects of responses to these psychostimulant medications. Conditioned place preference (CPP) for MP, AMPH and cocaine were evaluated in wild-type (WT) mice and their genetically engineered littermates, congenic on the C57Bl/6J background, that completely lack D4Rs (knockout or KO). In addition, the locomotor activity in these mice during the conditioning phase of CPP was tested in the CPP chambers. D4 receptor KO and WT mice showed CPP and increased locomotor activity in response to each of the three psychostimulants tested. D4R differentially modulates the CPP responses to MP, AMPH and cocaine. While the D4R genotype affected CPP responses to MP (high dose only) and AMPH (low dose only) it had no effects on cocaine. Inasmuch as CPP is considered an indicator of sensitivity to reinforcing responses to drugs these data suggest a significant but limited role of D4Rs in modulating conditioning responses to MP and AMPH. In the locomotor test, D4 receptor KO mice displayed attenuated increases in AMPH induced locomotor activity whereas responses to cocaine and MP did not differ. These results suggest distinct mechanisms for D4 receptor modulation of the reinforcing (perhaps via attenuating dopaminergic signalling) and locomotor properties of these stimulant drugs. Thus, individuals with D4 receptor polymorphisms might show enhanced reinforcing responses to MP and AMPH and attenuated locomotor response to AMPH. PMID- 19282422 TI - Involvement of PLC-beta3 in the effect of morphine on memory retrieval in passive avoidance task. AB - Phospholipase C (PLC) is one signalling effector enzyme whose activity is directly modulated by opioids. Several physiological studies have implicated PLC linked pathways in in-vivo pain regulation and opioid tolerance. Co administration of PLC-beta(2/3) activity blocker M119 with morphine resulted in a dramatic increase in morphine-induced amnesic effect in mice, proving a role for beta subunit of PLC enzyme in these processes. Administration of morphine to mice at amnesic dose increased PLC-beta(3) activity, with respect to basal value, in the membrane-soluble material from anterior cortex and hippocampal formation in brain areas. PLC-beta(3) appears to be simultaneously implicated in both analgesic and amnesic effects induced by administration of morphine to mice suggesting a commonality in the molecular mechanisms of morphine-induced analgesia and memory impairment. PMID- 19282423 TI - Effects of adjunctive reboxetine in patients with duloxetine-resistant depression: a 12-week prospective study. AB - The efficacy of the combination therapy with two antidepressants from different pharmacological families in patients with treatment-resistant depression has been reported in multiple studies. In this prospective 12-weeks open-label study, we assessed the effectiveness of the addition of reboxetine to 79 depressive outpatients diagnosed with major depressive disorder (MDD) according to the DSM IV criteria who had previously not responded, or had done so only in a partial way, over 8 weeks of conventional treatment, in monotherapy, with duloxetine. Efficacy was assessed using the 21-item Hamilton Depression Rating Scale (HDRS) and the Clinical Global Impression-Improvement (CGI-I). Safety was evaluated by recording spontaneously reported adverse events. Data were analysed on an intent to-treat basis, using the last-observation-carried-forward method. Mean HDRS reduction was 65.5% (P < 0.0001). The percentages of responders (>or=50% reduction in HDRS) and patients considered benefiting from complete remission (HDRS 25 mg/2 weeks (70% increase) and patients previously treated with clozapine (118% increase). Overall, number of hospital admissions did not increase, although those previously prescribed clozapine saw a 31% increase in admissions compared with patients not previously exposed to clozapine. This and other analyses of the same patient cohort indicate that RLAI produces most favourable outcomes in outpatients and those not previously treated with clozapine. PMID- 19282425 TI - Effects of ethanol and ecstasy on conditioned place preference in the rat. AB - The club drug ecstasy (3,4-methylenedioxymethylamphetamine or MDMA) is often taken recreationally with ethanol (EtOH). We have shown previously that EtOH potentiates the psychomotor effects of MDMA in rats. More recently, we demonstrated in striatal slices that MDMA produced preferential release of serotonin, but when combined with EtOH, the preferential release shifted to dopamine, raising the possibility that administration of EtOH may increase the reward effect of MDMA. To address this possibility, adult male Long-Evans rats were tested for conditioned place preference following treatment with saline, EtOH (0.75 g/kg), MDMA (6.6 mg/kg) or the combination. The only condition that produced a preference for the compartment associated with the drug was that of the drug combination. The current data are in line with anecdotal reports and one study in humans, indicating that EtOH alters the pharmacological effects of MDMA including self reports of enhanced or prolonged euphoria. Thus, administration of EtOH might increase the risk for compulsive use of MDMA, an issue that warrants further study. PMID- 19282426 TI - Elevated C-reactive protein in depression: a predictor of good long-term outcome with antidepressants and poor outcome with psychotherapy. PMID- 19282427 TI - Identification and functional analysis of pistil self-incompatibility factor HT-B of Petunia. AB - Gametophytic self-incompatibility (GSI) in Solanaceae, Rosaceae, and Plantaginaceae is controlled by a multiallelic S-locus. The specificities of pistil and pollen are controlled by separate S-locus genes, S-RNase and SLF/SFB, respectively. Although the S-specificity is determined by the S-locus genes, factors located outside the S-locus are also required for expression of GSI. HT-B is one of the pistil non-S-factors identified in Nicotiana and Solanum, and encodes a small asparagine/aspartate-rich extracellular protein with unknown biochemical function. Here, HT-B was cloned from Petunia and characterized. The structural features and expression pattern of Petunia HT-B were very similar to those of Nicotiana and Solanum. Unlike other solanaceous species, expression of HT-B was also observed in self-compatible Petunia species. RNA interference (RNAi)-mediated suppression of Petunia HT-B resulted in partial breakdown of GSI. Quantitative analysis of the HT-B mRNA accumulation in the transgenics showed that a 100-fold reduction is not sufficient and a >1000-fold reduction is required to achieve partial breakdown of GSI. PMID- 19282428 TI - Oral pravastatin prolongs survival time of scrapie-infected mice. AB - Statins are potent inhibitors of HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase in the cholesterol-biosynthesis pathway. They are either lipophilic (e.g. simvastatin) or hydrophilic [e.g. pravastatin (PRV)] compounds, considered mainly for long-term treatment of hypercholesterolaemic individuals. Beneficial effects of statins are not related exclusively to their lipid-lowering action; they also possess cholesterol-independent, pleiotropic effects (e.g. anti inflammatory and antioxidant). Recent studies revealed that simvastatin treatment increased survival significantly in scrapie-infected mice. Although PRV treatment results in measurable drug levels in the mouse brain, the anti-prion effect of this compound has not been investigated. Therefore, we aimed to test the potential therapeutic action of PRV in a murine scrapie model. Our study showed that high-dose and long-term oral PRV treatment prolonged survival times of strain 139A scrapie-infected mice significantly (194 versus 177 days) in the absence of any obvious toxicity, suggesting that protective effects of statins may be independent of absolute solvent or water solubility of the drug. PMID- 19282429 TI - Efficient production of infectious hepatitis C virus with adaptive mutations in cultured hepatoma cells. AB - Robust production of infectious hepatitis C virus (HCV) in cell culture was realized by using the JFH1 strain and the homologous chimeric J6/JFH1 strain in Huh-7.5 cells, a highly HCV-permissive subclone of Huh-7 cells. In this study, we aimed to establish a more efficient HCV-production system and to gain some insight into the adaptation mechanisms of efficient HCV production. By serial passaging of J6/JFH1-infected Huh-7.5 cells, we obtained culture-adapted J6/JFH1 variants, designated P-27, P-38 and P-47. Sequence analyses revealed that the adaptive mutant viruses P-27, P-38 and P-47 possessed eight mutations [four in E2, two in NS2, one in NS5A and one in NS5B), 10 mutations [two additional mutations in the 5'-untranslated region (5'-UTR) and core] and 11 mutations (three additional mutations in 5'-UTR, core and NS5B), respectively. We introduced amino acid substitutions into the wild-type J6/JFH1 clone, generated recombinant viruses with adaptive mutations and analysed their infectivity and ability to produce infectious viruses. The viruses with the adaptive mutations exhibited higher expression of HCV proteins than did the wild type in Huh-7.5 cells. Moreover, we provide evidence suggesting that the mutation N534H in the E2 glycoprotein of the mutant viruses conferred an advantage at the entry level. We thus demonstrate that an efficient HCV-production system could be obtained by introducing adaptive mutations into the J6/JFH1 genome. The J6/JFH1-derived mutant viruses presented here would be a good tool for producing HCV particles with enhanced infectivity and for studying the molecular mechanism of HCV entry. PMID- 19282430 TI - Genetic and epidemiological characterization of Stretch Lagoon orbivirus, a novel orbivirus isolated from Culex and Aedes mosquitoes in northern Australia. AB - Stretch Lagoon orbivirus (SLOV) was isolated in 2002 from pooled Culex annulirostris mosquitoes collected at Stretch Lagoon, near the Wolfe Creek national park in the Kimberley region of Western Australia. Conventional serological tests were unable to identify the isolate, and electron microscopy indicated a virus of the genus Orbivirus, family Reoviridae. Here, a cDNA subtraction method was used to obtain approximately one-third of the viral genome, and further sequencing was performed to complete the sequences of segment 1 (viral polymerase) and segment 2 (conserved inner-core protein). Phylogenetic analysis showed that SLOV should be considered a new species within the genus Orbivirus. A real-time RT-PCR test was designed to study the epidemiology of SLOV in the field. Six additional isolates of SLOV were identified, including isolates from four additional locations and two additional mosquito species. Horses, donkeys and goats were implicated as potential vertebrate hosts in a serological survey. PMID- 19282431 TI - Single amino acid changes in the mumps virus haemagglutinin-neuraminidase and polymerase proteins are associated with neuroattenuation. AB - It has previously been shown that three amino acid changes, one each in the fusion (F; Ala/Thr-91-->Thr), haemagglutinin-neuraminidase (HN; Ser-466-->Asn) and polymerase (L; Ile-736-->Val) proteins, are associated with attenuation of a neurovirulent clinical isolate of mumps virus (88-1961) following serial passage in vitro. Here, using full-length cDNA plasmid clones and site-directed mutagenesis, it was shown that the single amino acid change in the HN protein and to a lesser extent, the change in the L protein, resulted in neuroattenuation, as assessed in rats. The combination of both amino acid changes caused neuroattenuation of the virus to levels previously reported for the clinical isolate following attenuation in vitro. The amino acid change in the F protein, despite having a dramatic effect on protein function in vitro, was previously shown to not be involved in the observed neuroattenuation, highlighting the importance of conducting confirmatory in vivo studies. This report provides additional supporting evidence for the role of the HN protein as a virulence factor and, as far as is known, is the first report to associate an amino acid change in the L protein with mumps virus neuroattenuation. PMID- 19282432 TI - Evidence for modulation of BAG3 by polyomavirus JC early protein. AB - Polyomavirus JC (JCV) infects oligodendrocytes and astrocytes in the brain and is the cause of the demyelinating disease progressive multifocal leukoencephalopathy (PML). In cell culture, JCV infection is characterized by severe damage to cellular DNA, which begins early in infection, and a viral cytopathic effect, which is observed late in infection. Nevertheless, these JCV-infected cells show a low level of apoptosis, at both the early and late stages of infection. This suggests that there is conflicting interplay between viral anti-apoptotic pathways that seek to optimize virus production, e.g. through T antigen (T-Ag) p53 interaction, and cellular pro-apoptotic pathways that seek to eliminate virally infected cells. The apoptosis regulatory protein BAG3 is a member of the human Bcl-2-associated athanogene (BAG) family of proteins, which function as molecular co-chaperones through their interaction with Hsc70/Hsp70 and function in the regulation of the cellular stress response, proliferation and apoptosis. This study showed that BAG3 protein is downregulated upon JCV infection and that this effect is mediated by JCV T-Ag via repression of the BAG3 promoter. The site of action of T-Ag was mapped to an AP2 site in the BAG3 promoter, and gel shift and chromatin immunoprecipitation assays showed that T-Ag inhibited AP2 binding to this site, resulting in downregulation of BAG3 promoter expression. Using BAG3 and T-Ag expression and BAG3 siRNA, it was found that BAG3 and T-Ag had antagonistic effects on the induction of apoptosis, being anti-apoptotic and pro apoptotic, respectively. The significance of these interactions to the JCV life cycle is discussed. PMID- 19282433 TI - Binding of small interfering RNA molecules is crucial for RNA interference suppressor activity of rice hoja blanca virus NS3 in plants. AB - The NS3 protein of rice hoja blanca virus represents a viral suppressor of RNA interference (RNAi) that sequesters small interfering (si)RNAs in vitro. To determine whether this siRNA binding property is the critical determinant for the suppressor activity of NS3, NS3 was altered by alanine point mutations and the resulting mutant proteins were tested for both siRNA binding ability and RNAi suppressor activity in plants. Alanine substitutions of lysine residues at positions 173-175 resulted in mutant proteins that lost both their affinity for siRNAs and their RNAi suppressor activity in planta. This indicates that siRNA binding of NS3 is indeed essential for the suppressor function of NS3 and that residues at positions 173-175 are involved in the siRNA binding and suppressor activities. PMID- 19282434 TI - Vertical transmission of Prunus necrotic ringspot virus: hitch-hiking from gametes to seedling. AB - The aim of this work was to follow Prunus necrotic ringspot virus (PNRSV) infection in apricot reproductive tissues and transmission of the virus to the next generation. For this, an analysis of viral distribution in apricot reproductive organs was carried out at different developmental stages. PNRSV was detected in reproductive tissues during gametogenesis. The virus was always present in the nucellus and, in some cases, in the embryo sac. Studies within infected seeds at the embryo globular stage revealed that PNRSV infects all parts of the seed, including embryo, endosperm and testa. In the torpedo and bent cotyledon developmental stages, high concentrations of the virus were detected in the testa and endosperm. At seed maturity, PNRSV accumulated slightly more in the embryo than in the cotyledons. In situ hybridization showed the presence of PNRSV RNA in embryos obtained following hand-pollination of virus-free pistils with infected pollen. Interestingly, tissue-printing from fruits obtained from these pistils showed viral RNA in the periphery of the fruits, whereas crosses between infected pistils and infected pollen resulted in a total invasion of the fruits. Taken together, these results shed light on the vertical transmission of PNRSV from gametes to seedlings. PMID- 19282435 TI - Adenovirus serotype 5 infects human dendritic cells via a coxsackievirus adenovirus receptor-independent receptor pathway mediated by lactoferrin and DC SIGN. AB - The coxsackievirus-adenovirus receptor (CAR) is the described primary receptor for adenovirus serotype 5 (Ad5), a common human pathogen that has been exploited as a viral vector for gene therapy and vaccination. This study showed that monocytes and dendritic cells (DCs), such as freshly isolated human blood myeloid DCs, plasmacytoid DCs and monocyte-derived DCs, are susceptible to recombinant Ad5 (rAd5) infection despite their lack of CAR expression. Langerhans cells and dermal DCs from skin expressed CAR, but blocking CAR only partly decreased rAd5 infection, together suggesting that other receptor pathways mediate viral entry of these cells. Lactoferrin (Lf), an abundant protein in many bodily fluids known for its antiviral and antibacterial properties, promoted rAd5 infection in all cell populations except plasmacytoid DCs using a CAR-independent process. Lf caused phenotypic differentiation of the DCs, but cell activation played only a minor role in the increase in infection frequencies. The C-type lectin receptor DC-SIGN facilitated viral entry of rAd5-Lf complexes and this was dependent on high-mannose-type N-linked glycans on Lf. These results suggest that Lf present at high levels at mucosal sites can facilitate rAd5 attachment and enhance infection of DCs. A better understanding of the tropism and receptor mechanisms of Ad5 may help explain Ad5 pathogenesis and guide the engineering of improved rAd vectors. PMID- 19282436 TI - Errors in administration of parenteral drugs in intensive care units: multinational prospective study. AB - OBJECTIVE: To assess on a multinational level the frequency, characteristics, contributing factors, and preventive measures of administration errors in parenteral medication in intensive care units. DESIGN: Observational, prospective, 24 hour cross sectional study with self reporting by staff. SETTING: 113 intensive care units in 27 countries. PARTICIPANTS: 1328 adults in intensive care. MAIN OUTCOME MEASURES: Number of errors; impact of errors; distribution of error characteristics; distribution of contributing and preventive factors. RESULTS: 861 errors affecting 441 patients were reported: 74.5 (95% confidence interval 69.5 to 79.4) events per 100 patient days. Three quarters of the errors were classified as errors of omission. Twelve patients (0.9% of the study population) experienced permanent harm or died because of medication errors at the administration stage. In a multiple logistic regression with patients as the unit of analysis, odds ratios for the occurrence of at least one parenteral medication error were raised for number of organ failures (odds ratio per increase of one organ failure: 1.19, 95% confidence interval 1.05 to 1.34); use of any intravenous medication (yes v no: 2.73, 1.39 to 5.36); number of parenteral administrations (per increase of one parenteral administration: 1.06, 1.04 to 1.08); typical interventions in patients in intensive care (yes v no: 1.50, 1.14 to 1.96); larger intensive care unit (per increase of one bed: 1.01, 1.00 to 1.02); number of patients per nurse (per increase of one patient: 1.30, 1.03 to 1.64); and occupancy rate (per 10% increase: 1.03, 1.00 to 1.05). Odds ratios for the occurrence of parenteral medication errors were decreased for presence of basic monitoring (yes v no: 0.19, 0.07 to 0.49); an existing critical incident reporting system (yes v no: 0.69, 0.53 to 0.90); an established routine of checks at nurses' shift change (yes v no: 0.68, 0.52 to 0.90); and an increased ratio of patient turnover to the size of the unit (per increase of one patient: 0.73, 0.57 to 0.93). CONCLUSIONS: Parenteral medication errors at the administration stage are common and a serious safety problem in intensive care units. With the increasing complexity of care in critically ill patients, organisational factors such as error reporting systems and routine checks can reduce the risk for such errors. PMID- 19282437 TI - Practice based commissioning in the UK. PMID- 19282439 TI - What is the most effective management of neurogenic bladder dysfunction? PMID- 19282440 TI - Written informed consent and selection bias in observational studies using medical records: systematic review. AB - OBJECTIVES: To determine whether informed consent introduces selection bias in prospective observational studies using data from medical records, and consent rates for such studies. DESIGN: Systematic review. DATA SOURCES: Embase, Medline, and the Cochrane Library up to March 2008, reference lists from pertinent articles, and searches of electronic citations. STUDY SELECTION: Prospective observational studies reporting characteristics of participants and non participants approached for informed consent to use their medical records. Studies were selected independently in duplicate; a third reviewer resolved disagreements. DATA EXTRACTION: Age, sex, race, education, income, or health status of participants and non-participants, the participation rate in each study, and susceptibility of these calculations to threats of selection and reporting bias. RESULTS: Of 1650 citations 17 unique studies met inclusion criteria and had analysable data. Across all outcomes there were differences between participants and non-participants; however, there was a lack of consistency in the direction and the magnitude of effect. Of 161 604 eligible patients, 66.9% consented to use of data from their medical records. CONCLUSIONS: Significant differences between participants and non-participants may threaten the validity of results from observational studies that require consent for use of data from medical records. To ensure that legislation on privacy does not unduly bias observational studies using medical records, thoughtful decision making by research ethics boards on the need for mandatory consent is necessary. PMID- 19282441 TI - Screening for MCAD deficiency in newborns. PMID- 19282442 TI - Cases of acute and severe malnutrition in Somalia rise "exponentially". PMID- 19282443 TI - China tells doctors to quit smoking to set example to patients. PMID- 19282446 TI - The efficiency and fidelity of 8-oxo-guanine bypass by DNA polymerases delta and eta. AB - A DNA lesion created by oxidative stress is 7,8-dihydro-8-oxo-guanine (8-oxoG). Because 8-oxoG can mispair with adenine during DNA synthesis, it is of interest to understand the efficiency and fidelity of 8-oxoG bypass by DNA polymerases. We quantify bypass parameters for two DNA polymerases implicated in 8-oxoG bypass, Pols delta and eta. Yeast Pol delta and yeast Pol eta both bypass 8-oxoG and misincorporate adenine during bypass. However, yeast Pol eta is 10-fold more efficient than Pol delta, and following bypass Pol eta switches to less processive synthesis, similar to that observed during bypass of a cis-syn thymine thymine dimer. Moreover, yeast Pol eta is at least 10-fold more accurate than yeast Pol delta during 8-oxoG bypass. These differences are maintained in the presence of the accessory proteins RFC, PCNA and RPA and are consistent with the established role of Pol eta in suppressing ogg1-dependent mutagenesis in yeast. Surprisingly different results are obtained with human and mouse Pol eta. Both mammalian enzymes bypass 8-oxoG efficiently, but they do so less processively, without a switch point and with much lower fidelity than yeast Pol eta. The fact that yeast and mammalian Pol eta have intrinsically different catalytic properties has potential biological implications. PMID- 19282447 TI - Proofreading exonuclease activity of human DNA polymerase delta and its effects on lesion-bypass DNA synthesis. AB - Replicative DNA polymerases possess 3' --> 5' exonuclease activity to reduce misincorporation of incorrect nucleotides by proofreading during replication. To examine if this proofreading activity modulates DNA synthesis of damaged templates, we constructed a series of recombinant human DNA polymerase delta (Pol delta) in which one or two of the three conserved Asp residues in the exonuclease domain are mutated, and compared their properties with that of the wild-type enzyme. While all the mutant enzymes lost more than 95% exonuclease activity and severely decreased the proofreading activity than the wild-type, the bypass efficiency of damaged templates was varied: two mutant enzymes, D515V and D402A/D515A, gave higher bypass efficiencies on templates containing an abasic site, but another mutant, D316N/D515A, showed a lower bypass efficiency than the wild-type. All the enzymes including the wild-type inserted an adenine opposite the abasic site, whereas these enzymes inserted cytosine and adenine opposite an 8-oxoguanine with a ratio of 6:4. These results indicate that the exonuclease activity of human Pol delta modulates its intrinsic bypass efficiency on the damaged template, but does not affect the choice of nucleotide to be inserted. PMID- 19282448 TI - Novel method for high-throughput colony PCR screening in nanoliter-reactors. AB - We introduce a technology for the rapid identification and sequencing of conserved DNA elements employing a novel suspension array based on nanoliter (nl) reactors made from alginate. The reactors have a volume of 35 nl and serve as reaction compartments during monoseptic growth of microbial library clones, colony lysis, thermocycling and screening for sequence motifs via semi quantitative fluorescence analyses. nl-Reactors were kept in suspension during all high-throughput steps which allowed performing the protocol in a highly space effective fashion and at negligible expenses of consumables and reagents. As a first application, 11 high-quality microsatellites for polymorphism studies in cassava were isolated and sequenced out of a library of 20,000 clones in 2 days. The technology is widely scalable and we envision that throughputs for nl-reactor based screenings can be increased up to 100,000 and more samples per day thereby efficiently complementing protocols based on established deep-sequencing technologies. PMID- 19282449 TI - Distinctive sequence patterns in metazoan and yeast nucleosomes: implications for linker histone binding to AT-rich and methylated DNA. AB - Linker histones (LHs) bind to the DNA entry/exit points of nucleosomes and demonstrate preference for AT-rich DNA, although the recognized sequence patterns remain unknown. These patterns are expected to be more pronounced in metazoan nucleosomes with abundant LHs, compared to yeast nucleosomes with few LHs. To test this hypothesis, we compared the nucleosome core particle (NCP) sequences from chicken, Drosophila and yeast, extending them by the flanking sequences extracted from the genomes. We found that the known approximately 10-bp periodic oscillation of AT-rich elements goes beyond the ends of yeast nucleosomes, but is distorted in metazoan sequences where the 'out-of-phase' AT-peaks appear at the NCP ends. The observed difference is likely to be associated with sequence specific LH binding. We therefore propose a new structural model for LH binding to metazoan nucleosomes, postulating that the highly conserved nonpolar 'wing' region of the LH globular domain (tetrapeptide GVGA) recognizes AT-rich fragments through hydrophobic interactions with the thymine methyl groups. These interactions lead to DNA bending at the NCP ends and formation of a 'stem-like' structure. The same mechanism accounts for the high affinity of LH to methylated DNA-a feature critical for stabilization of the higher-order structure of chromatin and for repression of transcription. PMID- 19282450 TI - An archaeal Rad54 protein remodels DNA and stimulates DNA strand exchange by RadA. AB - Rad54 protein is a key member of the RAD52 epistasis group required for homologous recombination in eukaryotes. Rad54 is a duplex DNA translocase that remodels both DNA and protein-DNA complexes, and functions at multiple steps in the recombination process. Here we use biochemical criteria to demonstrate the existence of this important protein in a prokaryotic organism. The Sulfolobus solfataricus Rad54 (SsoRad54) protein is a double-strand DNA-dependent ATPase that can alter the topology of duplex DNA. Like its eukaryotic homolog, it interacts directly with the S. solfataricus Rad51 homologue, SsoRadA, to stimulate DNA strand exchange. Confirmation of this protein as an authentic Rad54 homolog establishes an essential phylogenetic bridge for identifying Rad54 homologs in the archaeal and bacterial domains. PMID- 19282451 TI - Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles. AB - We use cryo-electron microscopy (cryo-EM) to study the 3D shapes of 94-bp-long DNA minicircles and address the question of whether cyclization of such short DNA molecules necessitates the formation of sharp, localized kinks in DNA or whether the necessary bending can be redistributed and accomplished within the limits of the elastic, standard model of DNA flexibility. By comparing the shapes of covalently closed, nicked and gapped DNA minicircles, we conclude that 94-bp-long covalently closed and nicked DNA minicircles do not show sharp kinks while gapped DNA molecules, containing very flexible single-stranded regions, do show sharp kinks. We corroborate the results of cryo-EM studies by using Bal31 nuclease to probe for the existence of kinks in 94-bp-long minicircles. PMID- 19282452 TI - Detection of siRNA administered to cells and animals by using a fluorescence intensity distribution analysis polarization system. AB - Small interfering RNA (siRNA) has excellent pharmacological features and is expected to be used for therapeutic drug development. To this end, however, new RNA technology needs to be established so that extremely small amounts (less than 1 pmol) of siRNA can be detected in organs of experimental animals and in human blood to facilitate pharmacokinetics studies. An important feature is that this new technology is not dependent on radioisotopes and can detect siRNA molecules identical to those used for drug development in preclinical tests with experimental animals or in clinical tests with humans. We report a convenient method that can detect small amounts of siRNA. The method uses high-power confocal microscopic analysis of fluorescence polarization in DNA probes that are bound to one of the strands of siRNA and directly quantitates the copy number of siRNA molecule after extraction from specimens. A pharmacokinetic study to examine the blood retention time of siRNA/cationic liposomes in mice showed that this straightforward method is consistent with the other reverse transcriptase polymerase chain reaction amplification-based method. We believe that the entire process is simple and applicable for a high-throughput analysis, which provides excellent technical support for fundamental research on RNA interference and development of siRNA drugs. PMID- 19282453 TI - A large-scale chemical modification screen identifies design rules to generate siRNAs with high activity, high stability and low toxicity. AB - The use of chemically synthesized short interfering RNAs (siRNAs) is currently the method of choice to manipulate gene expression in mammalian cell culture, yet improvements of siRNA design is expectably required for successful application in vivo. Several studies have aimed at improving siRNA performance through the introduction of chemical modifications but a direct comparison of these results is difficult. We have directly compared the effect of 21 types of chemical modifications on siRNA activity and toxicity in a total of 2160 siRNA duplexes. We demonstrate that siRNA activity is primarily enhanced by favouring the incorporation of the intended antisense strand during RNA-induced silencing complex (RISC) loading by modulation of siRNA thermodynamic asymmetry and engineering of siRNA 3'-overhangs. Collectively, our results provide unique insights into the tolerance for chemical modifications and provide a simple guide to successful chemical modification of siRNAs with improved activity, stability and low toxicity. PMID- 19282454 TI - Protein hnRNP A1 and its derivative Up1 unfold quadruplex DNA in the human KRAS promoter: implications for transcription. AB - The promoter of the human KRAS proto-oncogene contains a structurally polymorphic nuclease hypersensitive element (NHE) whose purine strand forms a parallel G quadruplex structure (called 32R). In a previous work we reported that quadruplex 32R is recognized by three nuclear proteins: PARP-1, Ku70 and hnRNP A1. In this study we describe the interaction of recombinant hnRNP A1 (A1) and its derivative Up1 with the KRAS G-quadruplex. Mobility-shift experiments show that A1/Up1 binds specifically, and also with a high affinity, to quadruplex 32R, while CD demonstrates that the proteins strongly reduce the intensity of the 260 nm ellipticity-the hallmark for parallel G4-DNA-and unfold the G-quadruplex. Fluorescence resonance energy transfer melting experiments reveal that A1/Up1 completely abrogates the cooperative quadruplex-to-ssDNA transition that characterizes the KRAS quadruplex and facilitates the association between quadruplex 32R and its complementary polypyrimidine strand. When quadruplex 32R is stabilized by TMPyP4, A1/Up1 brings about only a partial destabilization of the G4-DNA structure. The possible role played by hnRNP A1 in the mechanism of KRAS transcription is discussed. PMID- 19282455 TI - Background dopamine concentration dependently facilitates long-term potentiation in rat prefrontal cortex through postsynaptic activation of extracellular signal regulated kinases. AB - Altered levels of tonic/background dopamine in prefrontal cortex (PFC) may underlie modifications of executive cognitive function. We showed previously in rat PFC slices that exogenously supplied background dopamine facilitates induction of long-term potentiation (LTP), a possible cellular substrate for the long-term component of executive cognitive function. In the present study, we characterized cellular and molecular mechanisms underlying this modulatory dopamine effect. We show first that the LTP-facilitating effect of tonic/background dopamine follows an inverted-U shape concentration curve and that the effective level of background dopamine slowly activates postsynaptic extracellular signal-regulated kinases (ERKs) to facilitate LTP. Furthermore, we show the evidence that LTP-inducing high-frequency stimulation evokes endogenous release of dopamine in PFC slices. This fast dopamine serves as a trigger for LTP in the presence of the background dopamine. In its absence, the endogenous dopamine triggered, instead, long-term depression. These results indicate that appropriate levels of tonic/background dopamine serve to activate critical molecular factors in PFC neurons and thereby facilitate induction of synaptic potentiation. PMID- 19282456 TI - No neglect-specific deficits in reaching tasks. AB - It is well established that patients with hemispatial neglect present with severe visuospatial impairments, but studies that have directly investigated visuomotor control have revealed diverging results, with some studies showing that neglect patients perform relatively better on such tasks. The present study compared the visuomotor performance of patients with and without neglect after right hemisphere stroke with those of age-matched controls. Participants were asked to point either directly towards targets or halfway between two stimuli, both with and without visual feedback during movement. Although we did not find any neglect specific impairment, both patient groups showed increased reaction times to leftward stimuli as well as decreased accuracies for open loop leftward reaches. We argue that these findings agree with the view that neglect patients code spatial parameters for action veridically. Moreover, we suggest that lesions in the right hemisphere may cause motor deficits irrespective of the presence of neglect and we performed an initial voxel-lesion symptom analysis to assess this. Lesion-symptom analysis revealed that the reported deficits did not result from damage to neglect-associated areas alone, but were further associated with lesions to crucial nodes in the visuomotor control network (the basal ganglia as well as occipito-parietal and frontal areas). PMID- 19282457 TI - Failure to achieve a complete hematologic response at the time of a major cytogenetic response with second-generation tyrosine kinase inhibitors is associated with a poor prognosis among patients with chronic myeloid leukemia in accelerated or blast phase. AB - Second-generation tyrosine kinase inhibitors are effective in Philadelphia chromosome-positive (Ph(+)) acute lymphoblastic leukemia (ALL) and chronic myeloid leukemia (CML). Occasionally, patients with Ph(+) ALL, or accelerated phase (AP) or blast phase (BP) CML achieve a major cytogenetic response (MCyR) but not a complete hematologic response (CHR). We analyzed 126 patients with CML in AP or BP, or with Ph(+) ALL treated with dasatinib or nilotinib after imatinib failure. Twenty patients received sequential treatment with both dasatinib and nilotinib for a total of 146 instances. CHR and MCyR rates were 54% and 37%, respectively in AP, 17% and 39% in BP, and 33% and 50% in Ph+ ALL. Failure to achieve a CHR at the time of achievement of a MCyR was associated with an inferior outcome, similar to that of patients without a MCyR (2-year survival rate, 37% and 35%, respectively). In contrast, patients with MCyR and concomitant CHR had a 77% 2-year survival rate. Twelve of 29 patients with MCyR without concomitant CHR later achieved a CHR; the 2-year survival rate for these patients was 55% compared with 22% for those who never achieved a CHR. These results suggest that achievement of a MCyR without concomitant CHR is associated with poor outcome. PMID- 19282458 TI - Increased signaling through p62 in the marrow microenvironment increases myeloma cell growth and osteoclast formation. AB - Adhesive interactions between multiple myeloma (MM) cells and marrow stromal cells activate multiple signaling pathways including nuclear factor kappaB (NF kappaB), p38 mitogen-activated protein kinase (MAPK), and Jun N-terminal kinase (JNK) in stromal cells, which promote tumor growth and bone destruction. Sequestosome-1 (p62), an adapter protein that has no intrinsic enzymatic activity, serves as a platform to facilitate formation of signaling complexes for these pathways. Therefore, we determined if targeting only p62 would inhibit multiple signaling pathways activated in the MM microenvironment and thereby decrease MM cell growth and osteoclast formation. Signaling through NF-kappaB and p38 MAPK was increased in primary stromal cells from MM patients. Increased interleukin-6 (IL-6) production by MM stromal cells was p38 MAPK-dependent while increased vascular cell adhesion molecule-1 (VCAM-1) expression was NF-kappaB dependent. Knocking-down p62 in patient-derived stromal cells significantly decreased protein kinase Czeta (PKCzeta), VCAM-1, and IL-6 levels as well as decreased stromal cell support of MM cell growth. Similarly, marrow stromal cells from p62(-/-) mice produced much lower levels of IL-6, tumor necrosis factor alpha (TNF-alpha), and receptor activator of NF-kappaB ligand (RANKL) and supported MM cell growth and osteoclast formation to a much lower extent than normal cells. Thus, p62 is an attractive therapeutic target for MM. PMID- 19282459 TI - Cancer in dyskeratosis congenita. AB - Dyskeratosis congenita (DC) is a rare inherited bone marrow failure syndrome. The spectrum of cancer susceptibility in this disorder of telomere biology has not been described. There were more than 500 cases of DC reported in the literature from 1910 to 2008; the National Cancer Institute (NCI) prospective DC cohort enrolled 50 cases from 2002 to 2007. Sixty cancers were reported in 52 literature cases, while 7 occurred among patients in the NCI DC cohort. The 2 cohorts were comparable in their median overall survival (42 years) and cumulative incidence of cancer (40%-50% by age 50 years). The most frequent solid tumors were head and neck squamous cell carcinomas (40% of patients in either cohort), followed by skin and anorectal cancer. The ratio of observed to expected cancers (O/E ratio) in the NCI cohort was 11-fold compared with the general population (P < .05). Significantly elevated O/E ratios were 1154 for tongue cancer and 195 for acute myeloid leukemia. Survival after bone marrow transplantation for aplastic anemia or leukemia was poor in both cohorts. The frequency and types of cancer in DC are surpassed only by those in Fanconi anemia (FA), indicating that FA and DC have similarly high risks of adverse hematologic and neoplastic events, and patients with these diseases should be counseled and monitored similarly. PMID- 19282461 TI - Syk to death. PMID- 19282460 TI - Role of GM-CSF signaling in cell-based tumor immunization. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a potent adjuvant in cancer vaccination; however, the specific role of endogenous GM-CSF remains unknown. We performed cell-based vaccination in 2 tumor models. First, we vaccinated C57BL/6 mice lacking either GM-CSF, IL-5, or beta-common chain (betac), a receptor subunit essential for GM-CSF and IL-5 signaling, with melanoma cells engineered to produce GM-CSF. Tumor vaccination was effective in both GM-CSF(-/-) and IL-5(-/-) mice, showing that protective immunization is independent of both endogenous cytokines. However, all betac(-/-) animals developed tumor. Loss of tumor immunity in betac(-/-) mice does not reflect global impairment in cell-mediated immunity, as contact hypersensitivity reaction to haptens is unaltered. The importance of tumor cell-derived GM-CSF was highlighted by recruitment of dendritic cells at the vaccination site in wild type, GM-CSF(-/-), and IL-5(-/-) but not in betac(-/-) mice. In the second model, vaccination with unmodified RENCA cells showed similar results with efficient immunization in BALB/c wild-type and GM-CSF(-/-), whereas all betac(-/-) animals died. Altogether, our results strongly suggest that although endogenous GM-CSF and IL-5 are not required to induce tumor immunity, signaling through betac receptor is critically needed for efficient cancer vaccination in both genetically modified GM-CSF-secreting tumor cells and a spontaneously immunogenic models. PMID- 19282462 TI - Might as well face it: MLL's addicted to HOX. PMID- 19282463 TI - Platelet antigen-induced regulation in ITP. PMID- 19282464 TI - Granulocyte/macrophage-colony-stimulating factor autoantibodies and myeloid cell immune functions in healthy subjects. AB - High levels of granulocyte/macrophage-colony-stimulating factor (GM-CSF) autoantibodies are thought to cause pulmonary alveolar proteinosis (PAP), a rare syndrome characterized by myeloid dysfunction resulting in pulmonary surfactant accumulation and respiratory failure. Paradoxically, GM-CSF autoantibodies have been reported to occur rarely in healthy people and routinely in pharmaceutical intravenous immunoglobulin (IVIG) purified from serum pooled from healthy subjects. These findings suggest that either GM-CSF autoantibodies are normally present in healthy people at low levels that are difficult to detect or that serum pooled for IVIG purification may include asymptomatic persons with high levels of GM-CSF autoantibodies. Using several experimental approaches, GM-CSF autoantibodies were detected in all healthy subjects evaluated (n = 72) at low levels sufficient to rheostatically regulate multiple myeloid functions. Serum GM CSF was more abundant than previously reported, but more than 99% was bound and neutralized by GM-CSF autoantibody. The critical threshold of GM-CSF autoantibodies associated with the development of PAP was determined. Results demonstrate that free serum GM-CSF is tightly maintained at low levels, identify a novel potential mechanism of innate immune regulation, help define the therapeutic window for potential clinical use of GM-CSF autoantibodies to treat inflammatory and autoimmune diseases, and have implications for the pathogenesis of PAP. PMID- 19282465 TI - Glucocorticoid replacement therapy and pharmacogenetics in Addison's disease: effects on bone. AB - Context Patients with primary adrenal insufficiency (Addison's disease) receive more glucococorticoids than the normal endogenous production, raising concern about adverse effects on bone. OBJECTIVE: To determine i) the effects of glucocorticoid replacement therapy on bone, and ii) the impact of glucocorticoid pharmacogenetics. DESIGN, SETTING AND PARTICIPANTS: A cross-sectional study of two large Addison's cohorts from Norway (n=187) and from UK and New Zealand (n=105). MAIN OUTCOME MEASURES: Bone mineral density (BMD) was measured; the Z scores represent comparison with a reference population. Blood samples from 187 Norwegian patients were analysed for bone markers and common polymorphisms in genes that have been associated with glucocorticoid sensitivity. RESULTS: Femoral neck BMD Z-scores were significantly reduced in the patients (Norway: mean -0.28 (95% confidence intervals (CI) -0.42, -0.16); UK and New Zealand: -0.21 (95% CI 0.36, -0.06)). Lumbar spine Z-scores were reduced (Norway: -0.17 (-0.36, +0.01); UK and New Zealand: -0.57 (-0.78, -0.37)), and significantly lower in males compared with females (P=0.02). The common P-glycoprotein (ABCB1) polymorphism C3435T was significantly associated with total BMD (CC and CT>TT P=0.015), with a similar trend at the hip and spine. CONCLUSIONS: BMD at the femoral neck and lumbar spine is reduced in Addison's disease, indicating undesirable effects of the replacement therapy. The findings lend support to the recommendations that 15 25 mg hydrocortisone daily is more appropriate than the higher conventional doses. A common polymorphism in the efflux transporter P-glycoprotein is associated with reduced bone mass and might confer susceptibility to glucocorticoid induced osteoporosis. PMID- 19282466 TI - Prognostic significance of circulating tumor cells in patients with metastatic colorectal cancer. AB - BACKGROUND: We demonstrated that circulating tumor cell (CTC) number at baseline and follow-up is an independent prognostic factor in metastatic colorectal cancer (mCRC). This analysis was undertaken to explore whether patient and treatment characteristics impact the prognostic value of CTCs. PATIENTS AND METHODS: CTCs were enumerated with immunomagnetic separation from the blood of 430 patients with mCRC at baseline and on therapy. Patients were stratified into unfavorable and favorable prognostic groups based on CTC levels of > or = 3 or <3 CTCs/7.5 ml, respectively. Subgroups were analyzed by line of treatment, liver involvement, receipt of oxaliplatin, irinotecan, or bevacizumab, age, and Eastern Cooperative Oncology Group performance status (ECOG PS). RESULTS: Seventy-one percent of deaths have occurred. Median follow-up for living patients is 25.8 months. For all patients, progression-free survival (PFS) and overall survival (OS) for unfavorable compared with favorable baseline CTCs is shorter (4.4 versus 7.8 m, P = 0.004 for PFS; 9.4 versus 20.6 m, P < 0.0001 for OS). In all patient subgroups, unfavorable baseline CTC was associated with inferior OS (P < 0.001). In patients receiving first- or second-line therapy (P = 0.003), irinotecan (P = 0.0001), having liver involvement (P = 0.002), >/=65 years (P = 0.0007), and ECOG PS of zero (P = 0.04), unfavorable baseline CTC was associated with inferior PFS. CONCLUSION: Baseline CTC count is an important prognostic factor within specific subgroups defined by treatment or patient characteristics. PMID- 19282467 TI - Effectiveness of organised versus opportunistic mammography screening. AB - BACKGROUND: Detailed comparison of effectiveness between organised and opportunistic mammography screening operating in the same country has seldom been carried out. PATIENTS AND METHODS: Prognostic indicators, as defined in the European Guidelines, were used to evaluate screening effectiveness in Switzerland. Matching of screening programmes' records with population-based cancer registries enabled to compare indicators of effectiveness by screening and detection modality (organised versus opportunistic screening, unscreened, interval cancers). Comparisons of prognostic profile were also drawn with two Swiss regions uncovered by service screening of low and high prevalence of opportunistic screening, respectively. RESULTS: Opportunistic and organised screening yielded overall little difference in prognostic profile. Both screening types led to substantial stage shifting. Breast cancer prognostic indicators were systematically more favourable in Swiss regions covered by a programme. In regions without a screening programme, the higher the prevalence of opportunistic screening, the better was the prognostic profile. CONCLUSIONS: Organised screening appeared as effective as opportunistic screening. Mammography screening has strongly influenced the stage distribution of breast cancer in Switzerland, and a favourable impact on mortality is anticipated. Extension of organised mammography screening to the whole of Switzerland can be expected to further improve breast cancer prognosis in a cost-effective way. PMID- 19282468 TI - Quality of life and disease-related symptoms in previously treated Japanese patients with non-small-cell lung cancer: results of a randomized phase III study (V-15-32) of gefitinib versus docetaxel. AB - BACKGROUND: This report describes quality of life (QoL) findings of a randomized study comparing gefitinib with docetaxel in patients with advanced/metastatic pretreated non-small-cell lung cancer. PATIENTS AND METHODS: This open-label, phase III study randomized 490 Japanese patients to gefitinib (250 mg/day) or docetaxel (60 mg/m(2)/3 weeks), with survival as the primary outcome. Preplanned QoL analyses included Functional Assessment of Cancer Therapy-Lung (FACT-L), Trial Outcome Index (TOI) and Lung Cancer Subscale (LCS) improvement rates, and mean change from baseline. RESULTS: Gefitinib showed statistically significant benefits over docetaxel in QoL improvement rates (FACT-L 23% versus 14%, P = 0.023; TOI 21% versus 9%, P = 0.002) and mean change from baseline score [mean treatment difference: FACT-L 3.72 points, 95% confidence interval (CI) 0.55-6.89, P = 0.022; TOI 4.31 points, 95% CI 2.13-6.49, P < 0.001], although differences did not meet the clinically relevant six-point change. There were no significant differences between treatments in LCS improvement rates (23% versus 20%, P = 0.562) or mean change from baseline score (0.63 points, 95% CI -0.07 to 1.34, P = 0.077). CONCLUSIONS: Gefitinib improved aspects of QoL over docetaxel, with superior objective response rate and a more favorable tolerability profile and no statistically significant difference in overall survival (although noninferiority was not statistically proven). PMID- 19282469 TI - Prescription of opioids in Italy: everything, but the morphine. PMID- 19282470 TI - The Cartographers toolbox: building bigger and better human protein interaction networks. AB - One of the greatest challenges of the post-genomic era is the construction of a more comprehensive human protein interaction map. While this process may take many years to complete, the development of stringent high throughput techniques and the emergence of complementary assays mean that the aim of building a detailed binary map of the human interactome is now a very realistic goal. In particular, methods which facilitate the analysis of large numbers of membrane protein interactions mean that it will be possible to construct more extensive networks, which in turn provide new insights into the functional connectivity between intra- and extra-cellular processes. This is important as many therapeutic strategies are designed to elicit effects via 'tractable' cell surface proteins. Therefore, the construction of maps depicting the complexity of trans-cellular communication networks will not only improve our understanding of physiological processes, it will also aid the design of rational therapeutic strategies, with fewer potential side effects. This review aims to provide a basic insight into the approaches currently being used to construct binary human protein interaction networks, with particular reference to newer techniques, which have the potential to extend network coverage and aid the conditional annotation of interactome-scale protein interaction maps. PMID- 19282471 TI - RGS4 polymorphisms associated with variability of cognitive performance in a family-based schizophrenia sample. AB - Polymorphisms of the gene encoding the regulator of G protein signaling, subtype 4 (RGS4), may be associated with schizophrenia. Among first-episode schizophrenia patients, they are also associated with dorsolateral prefrontal cortex (DLPFC) volume. The DLPFC is a key region that regulates heritable cognitive functions implicated in schizophrenia pathogenesis. To further understand the relationship of RGS4 variants to schizophrenia, we examined their associations with cognitive functions among schizophrenia patients and their relatives. We analyzed 31 multiplex, multigenerational Caucasian families with schizophrenia recruited on the basis of 2 affected first-degree relatives. All participants underwent a computerized neurocognitive battery that evaluates accuracy and speed (response time) of performance on abstraction/mental flexibility; attention; verbal, spatial, and face memory; and spatial ability. "Tag" single-nucleotide polymorphisms (SNPs) representing common polymorphisms were genotyped. Measured genotype analyses accounting for family relationships were performed using Sequential Oligogenic Linkage Analysis Routines. SNPs rs10917670 ("SNP1") and rs951439 ("SNP7") were associated with face memory speed (P = .0003) at a significance level that survived Bonferroni correction (P = .039). The same SNPs have earlier been reported to be associated with schizophrenia. There also were uncorrected associations with rs10917670 ("SNP1") and rs951439 ("SNP7") on face memory efficiency (P = .03) and verbal memory efficiency (P = 0.02), rs28757217 on abstraction/mental flexibility speed (P = .02) and verbal memory efficiency (P = .03), SNP18 (rs2661319) on spatial memory accuracy (P = 0.02) and face memory speed (P = .03). RGS4 polymorphisms are associated with variations in cognitive functions and contribute a small but statistically significant proportion of variance in a family-based sample. PMID- 19282472 TI - Reductions in the N1 and P2 auditory event-related potentials in first hospitalized and chronic schizophrenia. AB - The N1 auditory event-related potential (ERP) is reduced in chronic schizophrenia, as is the P2 to attended tones. N1 reduction may be endophenotypic for schizophrenia, being reduced in twins of schizophrenic patients and showing heritability. Results in family members, however, are equivocal, with abnormally small N1 (consistent with an endophenotype) and abnormally large N1 (inconsistent with an endophenotype) reported. P2 has been little studied in schizophrenia or family members. One crucial step in establishing endophenotypes is to rule out causal chronicity factors. We examined schizophrenia patients within 1 year of first hospitalization (most within 2 wk), chronically ill patients, and matched controls to examine N1 and P2 reductions and disease stage. Two active target detection oddball tasks were used, one with 97-dB tones against 70-dB white masking noise, the second with 97-dB tones without noise. Results from 8 samples are reported: first-hospitalized patients and matched controls and chronic patients and matched controls for the 2 tasks. N1 and P2 were measured from the standard stimuli. N1 and P2 were significantly reduced in chronic patients, as expected, and reduced in first-hospitalized patients. Because N1 and P2 are reduced even at the first hospitalization for schizophrenia, they may serve as viable electrophysiological endophenotypes for the disorder. However, deficit early in the disease is necessary but not sufficient to establish these ERPs as endophenotypes. Deficits must next be demonstrated in at least a subset of unaffected family members, a crucial criterion for an endophenotype. PMID- 19282473 TI - Fe65 is required for Tip60-directed histone H4 acetylation at DNA strand breaks. AB - Fe65 is a binding partner of the Alzheimer's beta-amyloid precursor protein APP. The possible involvement of this protein in the cellular response to DNA damage was suggested by the observation that Fe65 null mice are more sensitive to genotoxic stress than WT counterpart. Fe65 associated with chromatin under basal conditions and its involvement in DNA damage repair requires this association. A known partner of Fe65 is the histone acetyltransferase Tip60. Considering the crucial role of Tip60 in DNA repair, we explored the hypothesis that the phenotype of Fe65 null cells depended on its interaction with Tip60. We demonstrated that Fe65 knockdown impaired recruitment of Tip60-TRRAP complex to DNA double strand breaks and decreased histone H4 acetylation. Accordingly, the efficiency of DNA repair was decreased upon Fe65 suppression. To explore whether APP has a role in this mechanism, we analyzed a Fe65 mutant unable to bind to APP. This mutant failed to rescue the phenotypes of Fe65 null cells; furthermore, APP/APLP2 suppression results in the impairment of recruitment of Tip60-TRRAP complex to DNA double strand breaks, decreased histone H4 acetylation and repair efficiency. On these bases, we propose that Fe65 and its interaction with APP play an important role in the response to DNA damage by assisting the recruitment of Tip60-TRRAP to DNA damage sites. PMID- 19282474 TI - A new model for studying tissue-specific mdr1a gene expression in vivo by live imaging. AB - Multidrug resistance continues to be a major impediment to successful chemotherapy in cancer patients. One cause of multidrug resistance is enhanced expression of the mdr1 gene, but the precise factors and physiological conditions controlling mdr1 expression are not entirely known. To gain a better understanding of mdr1 transcriptional regulation, we created a unique mouse model that allows noninvasive bioimaging of mdr1 gene expression in vivo and in real time. The model uses a firefly luciferase (fLUC) gene inserted by homologous recombination into the murine mdr1a genetic locus. The inserted fLUC gene is preceded by a neo expression cassette flanked by loxP sites, so that Cre-mediated recombination is required to configure the fLUC gene directly under the control of the endogenous mdr1a promoter. We now demonstrate that the mdr1a.fLUC knock-in is a faithful reporter for mdr1a expression in naive animals, in which fLUC mRNA levels and luminescence intensities accurately parallel endogenous mdr1a mRNA expression. We also demonstrate xenobiotic-inducible regulation of mdr1a.fLUC expression in real time, in parallel with endogenous mdr1a expression, resulting in a more detailed understanding of the kinetics of mdr1a gene induction. This mouse model demonstrates the feasibility of using bioimaging coupled with Cre/loxP conditional knock-in to monitor regulated gene expression in vivo. It represents a unique tool with which to study the magnitude and kinetics of mdr1a induction under a variety of physiologic, pharmacologic, genetic, and environmental conditions. PMID- 19282475 TI - Molecular recognition and self-assembly special feature: discrete stack of an odd number of polarized aromatic compounds revealing the importance of net vs. local dipoles. AB - Three polarized aromatic guest molecules (pyrene-4,5-dione, 1) form a triple layered stack in the box-shaped cavity of an organic pillared coordination cage in water. The cavity size strictly limits the number of stacked planar guests but does not restrict guest orientation, and thus enables the study of discrete stacks of polarized guests and their preferred conformations. Crystallographic study shows that the guest molecules in the cavity are rotated 120 degrees with respect to each other, cancelling the net dipole moment rather than the local dipole moment. The unique conformation of a discrete, triple stack of 1 sharply contrasts to the standard head-to-tail conformation in infinite stacks of 1. PMID- 19282476 TI - Genome-wide discovery of functional transcription factor binding sites by comparative genomics: the case of Stat3. AB - The identification of direct targets of transcription factors is a key problem in the study of gene regulatory networks. However, the use of high throughput experimental methods, such as ChIP-chip and ChIP-sequencing, is limited by their high cost and strong dependence on cellular type and context. We developed a computational method for the genome-wide identification of functional transcription factor binding sites based on positional weight matrices, comparative genomics, and gene expression profiling. The method was applied to Stat3, a transcription factor playing crucial roles in inflammation, immunity and oncogenesis, and able to induce distinct subsets of target genes in different cell types or conditions. A newly generated positional weight matrix enabled us to assign affinity scores of high specificity, as measured by EMSA competition assays. Phylogenetic conservation with 7 vertebrate species was used to select the binding sites most likely to be functional. Validation was carried out on predicted sites within genes identified as differentially expressed in the presence or absence of Stat3 by microarray analysis. Twelve of the fourteen sites tested were bound by Stat3 in vivo, as assessed by Chromatin Immunoprecipitation, allowing us to identify 9 Stat3 transcriptional targets. Given its high validation rate, and the availability of large transcription factor-dependent gene expression datasets obtained under diverse experimental conditions, our approach appears to be a valid alternative to high-throughput experimental assays for the discovery of novel direct targets of transcription factors. PMID- 19282477 TI - Hepatitis C virus NS2 is a protease stimulated by cofactor domains in NS3. AB - Chronic infection with hepatitis C virus (HCV) affects 130 million people worldwide and is a major cause of liver cirrhosis and liver cancer. After translation of the HCV RNA genome into a polyprotein, 2 viral proteases process its non-structural protein (NS) region. While the essential chymotrypsin-like serine protease NS3-4A mediates all cleavages downstream of NS3, the NS2-3 cysteine protease catalyzes a vital cleavage at the NS2/3 site. Protease activity of NS2-3 has been described to require, besides NS2, the N-terminal 181 aa of NS3. The latter domain corresponds to the NS3 serine protease domain and contains a structural Zn(2+)-binding site with functional importance for both viral proteases. The catalytic triad of the NS2-3 protease resides in NS2; the role of the NS3 part in proteolysis remained largely undefined. Here we report a basal proteolytic activity for NS2 followed by only 2 amino acids of NS3. Basal activity could be dramatically enhanced by the NS3 Zn(2+)-binding domain (NS3 amino acids 81-213) not only in cis but also in trans which, however, required a more extended N-terminal part of NS3 downstream of NS2 in cis. Thus, this study defines for the first time (i) NS2 as a bona fide protease, (ii) NS3 as its regulatory cofactor, and (iii) functional subdomains in NS3 that cooperate in NS2 protease activation. These findings give new mechanistic insights into function and regulation of the NS2 protease and have important implications for the development of anti-HCV therapeutics. PMID- 19282478 TI - Enhancement of 1,25-dihydroxyvitamin D3-mediated suppression of experimental autoimmune encephalomyelitis by calcitonin. AB - The active form of vitamin D, 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], suppresses disease development in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS). However, complete disease prevention only occurs with doses that dramatically elevate serum calcium levels, thus limiting the usefulness of 1,25(OH)(2)D(3) as a potential MS therapeutic agent. Because calcitonin (CT) is believed to be released by hypercalcemia and has been shown to be anti-inflammatory, we examined whether suppression of EAE by 1,25(OH)(2)D(3) could be mediated either in part or entirely by CT. Continuous administration of pharmacological doses of CT did not prevent EAE. However, a combination of CT and a subtherapeutic dose of 1,25(OH)(2)D(3) additively suppressed EAE without causing hypercalcemia. Moreover, CT decreased the dose of 1,25(OH)(2)D(3) required for disease suppression. Our results suggest that CT may be a significant factor but cannot account entirely for 1,25(OH)(2)D(3)-mediated suppression of EAE. PMID- 19282479 TI - Thermodynamic equilibrium between blue and green copper sites and the role of the protein in controlling function. AB - A combination of spectroscopies and density functional theory calculations indicate that there are large temperature-dependent absorption spectral changes present in green nitrite reductases (NiRs) due to a thermodynamic equilibrium between a green and a blue type 1 (T1) copper site. The axial methionine (Met) ligand is unconstrained in the oxidized NiRs, which results in an enthalpically favored (DeltaH approximately 4.6 kcal/mol) Met-bound green copper site at low temperatures, and an entropically favored (TDeltaS approximately 4.5 kcal/mol, at room temperature) Met-elongated blue copper site at elevated temperatures. In contrast to the NiRs, the classic blue copper sites in plastocyanin and azurin show no temperature-dependent behavior, indicating that a single species is present at all temperatures. For these blue copper proteins, the polypeptide matrix opposes the gain in entropy that would be associated with the loss of the weak axial Met ligand at physiological temperatures by constraining its coordination to copper. The potential energy surfaces of Met binding indicate that it stabilizes the oxidized state more than the reduced state. This provides a mechanism to tune down the reduction potential of blue copper sites by >200 mV. PMID- 19282480 TI - Synthesis of programmable integrases. AB - Accurate modification of the 3 billion-base-pair human genome requires tools with exceptional sequence specificity. Here, we describe a general strategy for the design of enzymes that target a single site within the genome. We generated chimeric zinc finger recombinases with cooperative DNA-binding and catalytic specificities that integrate transgenes with >98% accuracy into the human genome. These modular recombinases can be reprogrammed: New combinations of zinc finger domains and serine recombinase catalytic domains generate novel enzymes with distinct substrate sequence specificities. Because of their accuracy and versatility, the recombinases/integrases reported in this work are suitable for a wide variety of applications in biological research, medicine, and biotechnology where accurate delivery of DNA is desired. PMID- 19282481 TI - Mapping dynamic memories of gradually changing objects. AB - Our brain is able to maintain a continuously updated memory representation of objects despite changes in their appearance over time (aging faces or objects, growing trees, etc.). Although this ability is crucial for cognition and behavior, it was barely explored. Here, we investigate this memory characteristic using a protocol emulating face transformation. Observers were presented with a sequence of faces that gradually transformed over many days, from a known face (source) to a new face (target), in presentations separated by other stimuli. This practice resulted in a drastic change in the memory and recognition of the faces. Although identification of the source and older face instances was reduced, recent face instances were increasingly identified as the source and rated as highly similar to the memory of the source. Using an object perturbation method, we estimated the corresponding memory shift, showing that memory patterns shifted from the source neighborhood toward the target. Our findings suggest that memory is updated to account for object changes over time while still keeping associations with past appearances. These experimental results are broadly compatible with a recently developed model of associative memory that assumes attractor dynamics with a learning rule facilitated by novelty, shown to hold when objects change gradually over short timescales. PMID- 19282482 TI - Regulation of DNMT1 stability through SET7-mediated lysine methylation in mammalian cells. AB - Inheritance of epigenetic information encoded by cytosine DNA methylation patterns is crucial for mammalian cell survival, in large part through the activity of the maintenance DNA methyltransferase (DNMT1). Here, we show that SET7, a known histone methyltransferase, is involved in the regulation of protein stability of DNMT1. SET7 colocalizes and directly interacts with DNMT1 and specifically monomethylates Lys-142 of DNMT1. Methylated DNMT1 peaks during the S and G(2) phases of the cell cycle and is prone to proteasome-mediated degradation. Overexpression of SET7 leads to decreased DNMT1 levels, and siRNA mediated knockdown of SET7 stabilizes DNMT1. These results demonstrate that signaling through SET7 represents a means of DNMT1 enzyme turnover. PMID- 19282483 TI - Prenatal care effectiveness and utilization in Brazil. AB - The impact of prenatal care use on birth outcomes has been understudied in South American countries. This study assessed the effects of various measures of prenatal care use on birth weight (BW) and gestational age outcomes using samples of infants born without and with common birth defects from Brazil, and evaluated the demand for prenatal care. Prenatal visits improved BW in the group without birth defects through increasing both fetal growth rate and gestational age, but prenatal care visits had an insignificant effect on BW in the group with birth defects when adjusting for gestational age. Prenatal care delay had no effects on BW in both infant groups but increased preterm birth risk in the group without birth defects. Inadequate care versus intermediate care also increased LBW risk in the group without birth effects. Quantile regression analyses revealed that prenatal care visits had larger effects at low compared with high BW quantiles. Several other prenatal factors and covariates such as multivitamin use and number of previous live births had significant effects on the studied outcomes. The number of prenatal care visits was significantly affected by several maternal health and fertility indicators. Significant geographic differences in utilization were observed as well. The study suggests that more frequent use of prenatal care can increase BW significantly in Brazil, especially among pregnancies that are uncomplicated with birth defects but that are at high risk for low birth weight. Further research is needed to understand the effects of prenatal care use for pregnancies that are complicated with birth defects. PMID- 19282484 TI - Achieving measles control: lessons from the 2002-06 measles control strategy for Uganda. AB - BACKGROUND The 2002-06 measles control strategy for Uganda was implemented to strengthen routine immunization, undertake large-scale catch-up and follow-up vaccination campaigns, and to initiate nationwide case-based, laboratory-backed measles surveillance. This study examines the impact of this strategy on the epidemiology of measles in Uganda, and the lessons learnt. METHODS Number of measles cases and routine measles vaccination coverage reported by each district were obtained from the National Health Management Information System reports of 1997 to 2007. The immunization coverage by district in a given year was calculated by dividing the number of children immunized by the projected population in the same age category. Annual measles incidence for each year was derived by dividing the number of cases in a year by the mid-year projected population. Commercial measles IgM enzyme-linked immunoassay kits were used to confirm measles cases. RESULTS Routine measles immunization coverage increased from 64% in 1997 to 90% in 2004, then stabilized around 87%. The 2003 national measles catch-up and 2006 follow-up campaigns reached 100% of children targeted with a measles supplemental dose. Over 80% coverage was also achieved with other child survival interventions. Case-based measles surveillance was rolled out nationwide to provide continuous epidemiological monitoring of measles occurrence. Following a 93% decline in measles incidence and no measles deaths, epidemic resurgence of measles occurred 3 years after a measles campaign targeting a wide age group, but no indigenous measles virus (D(10)) was isolated. Recurrence was delayed in regions where children were offered an early second opportunity for measles vaccination. CONCLUSION The integrated routine and campaign approach to providing a second opportunity for measles vaccination is effective in interrupting indigenous measles transmission and can be used to deliver other child survival interventions. Measles control can be sustained and the inter-epidemic interval lengthened by offering an early second opportunity for measles vaccination through other health delivery strategies. PMID- 19282485 TI - The learning and maintenance of local vectors in desert ant navigation. AB - The desert ant Cataglyphis fortis has at least three types of navigational strategy that can guide it between its nest and a familiar food site. The initial strategy after first finding a food site is based on a path integration memory of the position of the food site with respect to the nest. A second strategy is based on visual snapshot memories of features viewed from near or on the way to the food site. A third strategy uses local vector memories of the direction and length of habitual route segments. We show here that while such local vectors encode sufficient information to guide an individual along both the direction and distance of a route segment, its acquisition and long-term maintenance requires support from the other two strategies. We trained ants along an L-shaped route, designed to show that ants can learn local vectors on the way to a food site. The sharp turn appears to present particular difficulties for the ants. When low bushes 20-30 m from the route were removed, local vectors were briefly unaffected, but then deteriorated. The vectors improved again once the missing bushes were replaced by artificial landmarks. The fragility of local vector memories may permit an ant the flexibility to adapt its route to fluctuations in the distribution of its resources. PMID- 19282486 TI - Local and global navigational coordinate systems in desert ants. AB - While foraging, the desert ant Cataglyphis fortis keeps track of its position with respect to its nest through a process of path integration (PI). Once it finds food, it can then follow a direct home vector to its nest. Furthermore, it remembers the coordinates of a food site, and uses these coordinates to return to the site. Previous studies suggest, however, that it does not associate any coordinates remembered from previous trips with familiar views such that it can produce a home vector when displaced to a familiar site. We ask here whether a desert ant uses any association between PI coordinates and familiar views to ensure consistent PI coordinates as it travels along a habitual route. We describe an experiment in which we manipulated the PI coordinates an ant has when reaching a distinctive point along a habitual route on the way to a feeder. The subsequent home vectors of the manipulated ants, when displaced from the food site to a test ground, show that also when a route memory is evoked at a significant point on the way to a food site, C. fortis does not reset its PI coordinates to those it normally has at that point. We use this result to argue that local vector memories, which encode the metric properties of a segment of a habitual route, must be encoded in a route-based coordinate system that is separate from the nest-based global coordinates. We propose a model for PI-based guidance that can account for several puzzling observations, and that naturally produces the route-based coordinate system required for learning and following local vectors. PMID- 19282487 TI - The basis of vagal efferent control of heart rate in a neotropical fish, the pacu, Piaractus mesopotamicus. AB - The role of the parasympathetic nervous system, operating via the vagus nerve, in determining heart rate (f(H)) and cardiorespiratory interactions was investigated in the neotropical fish Piaractus mesopotamicus. Motor nuclei of branches of cranial nerves VII, IX and X, supplying respiratory muscles and the heart, have an overlapping distribution in the brainstem, while the Vth motor nucleus is more rostrally located. Respiration-related efferent activity in the cardiac vagus appeared to entrain the heart to ventilation. Peripheral stimulation of the cardiac vagus with short bursts of electrical stimuli entrained the heart at a ratio of 1:1 over a range of frequencies, both below and sometimes above the intrinsic heart rate. Alternatively, at higher bursting frequencies the induced f(H) was slower than the applied stimulus, being recruited by a whole number fraction (1:2 to 1:6) of the stimulus frequency. These effects indicate that respiration-related changes in f(H) in pacu are under direct, beat-to-beat vagal control. Central burst stimulation of respiratory branches of cranial nerves VII, IX and X also entrained the heart, which implies that cardiorespiratory interactions can be generated reflexly. Central stimulation of the Vth cranial nerve was without effect on heart rate, possibly because its central projections do not overlap with cardiac vagal preganglionic neurons in the brainstem. However, bursts of activity recorded from the cardiac vagus were concurrent with bursts in this nerve, suggesting that cardiorespiratory interactions can arise within the CNS, possibly by irradiation from a central respiratory pattern generator, when respiratory drive is high. PMID- 19282488 TI - Responses of hatchling Xenopus tadpoles to water currents: first function of lateral line receptors without cupulae. AB - At later stages in larval life and also as adults, Xenopus can respond to water currents detected by their lateral-line sensory system. We have investigated when responses to water currents first appear and whether the first lateral line neuromasts operate in the same way as the adult organs. Just before and after hatching from their egg membranes we show that Xenopus embryos and tadpoles can respond to water currents by swimming into them. Local stimulation in immobilised animals where motor activity was recorded electrically suggested that the receptors detecting water currents were located between the eyes and the gills and were innervated by cranial nerves. In behaving tadpoles, responses to water currents were reduced following skin abrasion caudal to the eyes or treatment with neomycin, which is known to block hair cell function. We therefore used scanning electron microscopy to establish that rows of lateral line neuromasts with hair cells and kinocilia are present just caudal to the eyes at these stages of development. However, careful observations and manipulations of the kinocilia of neuromasts in living tadpoles failed to find any evidence that kinocilia were embedded in a jelly-like cupula. We conclude that, when they first start to function, these early neuromasts detect water movements which directly move their freely exposed hair cell kinocilia projecting out from the skin surface. Possible behavioural roles for the tadpoles responses to water are discussed. PMID- 19282489 TI - Novel neural correlates of operant conditioning in normal and differentially reared Lymnaea. AB - The aerial respiratory behaviour of the mollusc Lymnaea stagnalis is an important homeostatic behaviour that can be operantly conditioned. The central pattern generator underlying this behaviour, as well as motorneurons innervating the respiratory orifice, the pneumostome, have been identified and their activity can be monitored in the semi-intact preparation using electrophysiological recordings. In this study, we used both intact animals and semi-intact preparations to identify novel changes in the respiratory central pattern generator following operant conditioning. In addition, we reared animals in the absence of this respiratory behaviour throughout development, to investigate whether previous experience and activity-dependent plasticity during development are essential to allow neural plasticity in the adult. We found that animals raised normally (allowed to perform aerial respiratory behaviour) exhibited the expected reduction in aerial respiratory behaviour following operant conditioning. Then, using the semi-intact preparation, we identified novel neural changes within the network as a result of the conditioning. These included specific changes at the level of the central pattern generator interneurons, as well as the motor output. In the differentially reared intact animals, there was no behavioural reduction as a result of operant conditioning, although their baseline respiratory behaviour was already significantly reduced as a result of their differential rearing. There were, however, significant differences found in the network parameters in the semi-intact preparation, similar to those observed in normally reared animals. We thus provide evidence for neural plasticity within the network in the absence of significant behavioural changes in differentially reared animals, and show that plasticity was not dependent on previous activity of the network during development. PMID- 19282490 TI - Nervous and humoral control of cardiac performance in the winter flounder (Pleuronectes americanus). AB - Previous studies have suggested that flatfish lack adrenergic cardiac innervation and have a limited humoral adrenergic stress response. However, data on neurohormonal control of flatfish cardiac function is scarce, and has never been directly studied in vivo. Hence, we (1) injected neural and humoral antagonists into flounder (Pleuronectes americanus) in vivo to determine the contribution of autonomic innervation and circulating catecholamines to the control of resting cardiac function; (2) measured pre- and post-stress (90 s chase) catecholamine levels in this species; and (3) constructed in vivo catecholamine dose-response curves for cardiovascular function based on the results of the second experiment. In addition, we quantified the density (B(max)) and ligand-binding affinity (Kd) of flounder ventricular cell-surface beta-adrenoreceptors, and established whether they were of beta1 or beta2 subtype using pharmacological antagonists. The cholinergic contribution to resting flounder heart rate was comparable to other teleosts (cholinergic tonus 26%). Interestingly, however, bretylium increased heart rate, resulting in a negative resting adrenergic tonus (-11.9%), and we were unable to demonstrate that catecholamines supported cardiac function at rest or at circulating concentrations approximating those following an exhaustive chase (adrenaline, 21 nmol l(-1); noradrenaline, 14 nmol l(-1)). Myocardial B(max) was very high in the flounder (252.8 fmol mg(-1) protein), and it appears that flounder ventricular beta-adrenoreceptors are predominantly of the beta2 subtype [based on the inability of atenolol to displace [3H]CGP from the beta-adrenoreceptors, and the IC(50) value for ICI 118551 (1.91 x 10(-6) mol l(-1))]. However, the extremely low affinity (Kd 1.02 nmol l(-1)) for these receptors raises the possibility that the flounder heart is also populated by beta3-adrenoreceptors. PMID- 19282491 TI - Bats go head-under-heels: the biomechanics of landing on a ceiling. AB - Bats typically roost head-under-heels but they cannot hover in this position, thus, landing on a ceiling presents a biomechanical challenge. To land, a bat must perform an acrobatic flip that brings the claws of the toes in contact with the ceiling and do so gently enough as to avoid injury to its slender hindlimbs. In the present study, we sought to determine how bats land, to seek a link between landing kinematics and ceiling impact forces, and to determine whether landing strategies vary among bat species. To do this, we measured the kinematics and kinetics of landing behaviour in three species of bats as they landed on a force-measuring platform (Cynopterus brachyotis, N=3; Carollia perspicillata, N=5; Glossophaga soricina, N=5). Kinematics were similar for all bats within a species but differed among species. C. brachyotis performed four-point landings, during which body pitch increased until the ventral surface of the body faced the ceiling and the thumbs and hindlimbs simultaneously grasped the surface. Bats of the other two species performed two-point landings, whereby only the hindlimbs made contact with the ceiling. During these two-point landings, the hindlimbs were drawn up the side of the body to come in contact with the ceiling, causing simultaneous changes in body pitch, roll and yaw over the course of the landing sequence. Right-handed and left-handed forms of the two-point landing were observed, with individuals often switching back and forth between them among landing events. The four-point landing of C. brachyotis resulted in larger peak forces (3.7+/-2.4 body weights; median +/- interquartile range) than the two point landings of C. perspicillata (0.8+/-0.6 body weights) or G. soricina (0.8+/ 0.2 body weights). Our results demonstrate that the kinematics and kinetics of landing vary among bat species and that there is a correlation between the way a bat moves its body when it lands and the magnitude of peak impact force it experiences during that landing. We postulate that these interspecific differences in impact force could result because of stronger selective pressure for gentle landing in cave-roosting (C. perspicillata, G. soricina) versus foliage-roosting (C. brachyotis) species. PMID- 19282492 TI - Nitric oxide increases myocardial efficiency in the hypoxia-tolerant turtle Trachemys scripta. AB - Nitric oxide (NO) may influence cardiac mechanical performance relative to O2 consumption by depressing respiration rate and by affecting the excitation contraction coupling. Such effects of NO should be particularly important during hypoxia in species such as the hypoxia-tolerant turtle Trachemys scripta. In heart ventricle preparations from this species, the ratio of twitch force to O2 consumption increased by approximately 15% during full oxygenation and by approximately 60% during hypoxia in the presence of added L-arginine [the substrate for nitric oxide synthase (NOS)]. This effect was primarily due to a decrease in O2 consumption and may represent an increase in the twitch force obtained per ATP and/or in the ATP obtained per O2. Lactate production during hypoxia did not differ between preparations treated with either L-arginine or asymmetric dimethylarginine (ADMA), an inhibitor of NOS, suggesting that NO does not elicit a compensatory increase in anaerobic metabolism. ADMA did not reverse the effects of L-arginine on O2 consumption significantly, although pre-treatment with ADMA abolished the effect of L-arginine, consistent with the competitive binding of L-arginine and ADMA to NOS. Histochemical studies using the fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA) revealed NO production in the presence of added L-arginine. In conclusion, NO may augment heart contractility obtained per O2 by deceasing O2 consumption without affecting either lactate production or developed force. This effect was particularly pronounced under O2 deficiency and may therefore contribute towards preserving cardiac function and to the overall excellent hypoxic tolerance of the turtle. PMID- 19282493 TI - Mechanical and biological consequences of repetitive loading: crack initiation and fatigue failure in the red macroalga Mazzaella. AB - On rocky shores, wave-swept macroalgae experience dramatic and repeated wave induced hydrodynamic forces. However, previous studies of macroalgal mechanics have shown that individual waves are not forceful enough to account for observed rates of breakage. Instead, fatigue may contribute to algal breakage, with damage accumulating over time in conditions of repeated loading. Here I examine the entire process of fatigue, from crack initiation to eventual specimen fracture, in the common red alga Mazzaella. Propensity for fatigue failure in laboratory tests varied with life history phase and species: at a given repeated loading stress, male gametophytes endured more loading cycles before breakage than tetrasporophytes, which in turn lasted longer than female gametophytes; likewise, M. splendens withstood more loading cycles at a given repeated loading stress than M. flaccida. Fatigue failure begins with formation of cracks, the timing and location of which were assessed. Cracks formed, on average, after approximately 80-90% of cycles required for failure had passed, although crack timing varied with life history phase. Also, crack formation frequently occurred in association with endophytes and female gametophyte reproductive structures, suggesting a cost of endophyte infection and a tradeoff between reproduction and mechanical survival. Comparison between laboratory and field loading conditions provides robust confirmation that fatigue breaks fronds in natural M. flaccida populations. Large, female gametophyte fronds are predicted to be most susceptible to fatigue failure in the field, whereas small, male gametophyte fronds are least likely to break. PMID- 19282494 TI - Changes in efficiency and myosin expression in the small-muscle phenotype of mice selectively bred for high voluntary running activity. AB - Mice from lines selectively bred for high levels of voluntary wheel running express a high incidence of a small muscle phenotype ('mini-muscles') that may confer an adaptive advantage with respect to endurance-running capacity. Plantar flexors in the mini-muscle phenotype exhibit a high capacity for aerobic activity, including altered enzyme activities, loss of expression of type IIb myosin heavy chain (MHC), increased expression of type I, IIx and IIa MHC, and mechanical performance consistent with slower, more fatigue-resistant muscles. We hypothesized that these changes may accompany enhanced efficiency of contraction, perhaps in support of the enhanced capacity for endurance running. To assess efficiency, we measured work and associated oxygen consumption from isolated soleus and medial gastrocnemius muscles from mice with mini-muscle and normal phenotypes. We also measured the MHC expression of the plantar flexor muscles to better understand the physiological basis of any differences in efficiency. The proportion of the various MHC isoforms in the soleus was shifted toward a slightly faster phenotype in the mini-muscle mice, whereas in the gastrocnemius and plantaris it was shifted toward a markedly slower phenotype, with large reductions in type IIb MHC and large increases in type I, IIa, and IIx MHC. Soleus muscles from normal and mini-muscle mice showed no statistical differences in efficiency, but medial gastrocnemius from mini-muscle mice were significantly less efficient than those from normal mice, despite the distinctly slower MHC phenotype in mini-muscle mice. Thus, based on measures of efficiency from isolated muscles under conditions near optimal for power output, the shift toward a slower phenotype in 'mini' gastrocnemius muscles does not appear to confer advantages directly through increased efficiency. Rather, the slower phenotype may reduce energy used by the muscles and be permissive to enhanced running ability, perhaps by reducing reliance on anaerobic metabolism. PMID- 19282495 TI - T3 and the thyroid hormone beta-receptor agonist GC-1 differentially affect metabolic capacity and oxidative damage in rat tissues. AB - We compared the changes in tissue aerobic metabolism and oxidative damage elicited by hypothyroid rat treatment with T3 and its analog GC-1. Aerobic capacities, evaluated by cytochrome oxidase activities, were increased more by T3 than by GC-1. Furthermore, the response of the tissues to T3 was similar, whereas the response to GC-1 was high in liver, low in muscle and scarce in heart. Both treatments induced increases in ADP-stimulated O2 consumption, which were consistent with those in aerobic capacities. However, unlike T3, GC-1 differentially affected pyruvate/malate- and succinate-supported respiration, suggesting that respiratory chain components do not respond as a unit to GC-1 stimulation. According to the positive relationship between electron carrier levels and rates of mitochondrial generation of oxidative species, the most extensive damage to lipids and proteins was found in T3-treated rats. Examination of antioxidant enzyme activities and scavenger levels did not clarify whether oxidative damage extent also depended on different antioxidant system effectiveness. Conversely, the analysis of parameters determining tissue susceptibility to oxidants showed that pro-oxidant capacity was lower in GC-1- than in T3-treated rats, while antioxidant capacity was similar in treatment groups. Interestingly, both agonists decreased serum cholesterol levels, but only GC-1 restored euthyroid values of heart rate and indices of tissue oxidative damage, indicating that GC-1 is able to lower cholesterolemia, bypassing detrimental effects of T3. PMID- 19282496 TI - Impaired tactile learning is related to social role in honeybees. AB - Aging is commonly accompanied by a decline in cognitive functions such as learning and memory. In social insects, aging is tightly linked to social role. The honeybee (Apis mellifera L.) offers the unique opportunity to separate chronological age from social role. In the present paper, we tested whether chronological age, social role and the duration of performing this role affect tactile learning in honeybees. We compared acquisition, retention and discrimination between foragers with short and long foraging durations and age matched nurse bees. Our data show that chronological age is of minor importance for tactile learning, retention and discrimination whereas social role has a decisive impact. Tactile acquisition is severely impaired in bees that have foraged for more than two weeks but not in nurse bees of the same chronological age. Interestingly, neither discrimination nor retention appear to be impaired by long foraging duration. The complex associations between acquisition, discrimination and retention in bees of different social roles open up rich possibilities for future studies on the neuronal correlates of behavioural performance and underline that the honeybee has great potential as a model system in the biology of aging. PMID- 19282497 TI - Morphological and functional classification of ion-absorbing mitochondria-rich cells in the gills of Mozambique tilapia. AB - To clarify ion-absorbing functions and molecular mechanisms of mitochondria-rich (MR) cells, Mozambique tilapia (Oreochromis mossambicus) were acclimated to artificial freshwaters with normal or lowered Na+ and/or Cl- concentration: (1) normal Na+/normal Cl- (control); (2) normal Na+/low Cl-; (3) low Na+/normal Cl-; and (4) low Na+/low Cl-. Scanning electron microscopy (SEM) revealed that concave and convex apical surfaces of MR cells predominantly developed in low Na+ and low Cl- waters, respectively, whereas small apical pits predominated in control conditions. Expression of Na+/H+ exchanger-3 (NHE3) mRNA in the gills was increased in low Na+ waters (low Na+/normal Cl- and low Na+/low Cl-, whereas that of Na+/Cl- cotransporter (NCC) expression was upregulated in low Cl-, but not in low Na+/low Cl-. Immunofluorescence staining showed that enlarged NHE3 immunoreactive apical regions were concave or flat in low Na+ waters, whereas NCC immunoreactive regions were enlarged convexly in low Cl- waters. Using SEM immunocytochemistry the distribution of NHE3/NCC was compared with SEM images obtained simultaneously, it was further demonstrated that NHE3 and NCC were confined to concave and convex apical surfaces, respectively. These results indicated that small apical pits developed into concave apical surfaces to facilitate Na+ uptake through NHE3, and into convex apical surfaces to enhance Na+/Cl- uptake through NCC. Our findings integrated morphological and functional classifications of ion-absorbing MR cells in Mozambique tilapia. PMID- 19282498 TI - Acoustic scanning of natural scenes by echolocation in the big brown bat, Eptesicus fuscus. AB - Echolocation allows bats to orient and localize prey in complete darkness. The sonar beam of the big brown bat, Eptesicus fuscus, is directional but broad enough to provide audible echo information from within a 60-90 deg. cone. This suggests that the big brown bat could interrogate a natural scene without fixating each important object separately. We tested this idea by measuring the directional aim and duration of the bat's sonar beam as it performed in a dual task, obstacle avoidance and insect capture. Bats were trained to fly through one of two openings in a fine net to take a tethered insect at variable distances behind the net. The bats sequentially scanned the edges of the net opening and the prey by centering the axis of their sonar beam with an accuracy of approximately 5 deg. The bats also shifted the duration of their sonar calls, revealing sequential sampling along the range axis. Changes in duration and directional aim were correlated, showing that the bat first inspected the hole, and then shifted its gaze to the more distant insect, before flying through the net opening. Contrary to expectation based on the sonar beam width, big brown bats encountering a complex environment accurately pointed and shifted their sonar gaze to sequentially inspect closely spaced objects in a manner similar to visual animals using saccades and fixations to scan a scene. The findings presented here from a specialized orientation system, echolocation, offer insights into general principles of active sensing across sensory modalities for the perception of natural scenes. PMID- 19282499 TI - Movement encoding by a stretch receptor in the soft-bodied caterpillar, Manduca sexta. AB - In a wide variety of animals, stretch receptors provide proprioceptive feedback for motion control. However, for animals that lack a stiff skeleton, it is unclear what information is being detected and how this is incorporated into behavior. Because such animals can change their body shape from moment-to-moment, information about body configuration could be particularly important for coordination. This study uses larval stage Lepidoptera (Manduca sexta) to examine how the longitudinal stretch receptor organ (SRO) responds to behaviorally appropriate movements. We characterized the responses of the SRO to changes in strain using magnitudes and velocities matching those seen physiologically. We found that the SRO response characteristics are compatible with the regulation of stance and with the defensive response to noxious stimuli. However, we also found that movements during crawling produce SRO responses that are dominated by the interdependence of phasic, tonic and slowly adaptive components. Ablation of stretch receptors in the proleg-bearing, fourth abdominal segment did not have any observable effect on behaviors, which suggests that the SROs are not essential for coordinating overt movements. We discuss the implications of these findings in the context of specific behaviors, and explore how the SRO response might be utilized during animal behavior. PMID- 19282500 TI - Mechanisms of food provisioning of honeybee larvae by worker bees. AB - Although it has clearly been demonstrated in previous studies that honeybees inspect their worker brood in a non-random fashion, it is still unclear which signals and cues worker bees use to monitor the nutritional state of their brood. Here we show that worker bees can recognize and quantify the larval food present in a brood cell olfactorily and identify potential mechanical signals produced by the brood. There is no evidence for additional chemical hunger signals produced by the larvae. However, the pattern of movement of larvae within their cells changes with their nutritional state and might provide additional information to nurse bees. PMID- 19282501 TI - Function of the extrinsic hindlimb muscles in trotting dogs. AB - The extrinsic appendicular muscles of mammals have been suggested to impose parasagittal torques on the trunk that require recruitment of the oblique hypaxial muscles for stabilization. To determine if the recruitment of the protractors and retractors of the hindlimb are compatible with this hypothesis, we monitored changes in the recruitment of eleven muscles that span the hip joint to controlled manipulations of locomotor forces in trotting dogs. The results indicate that the primary retractor muscles of the hindlimb produce a small retraction moment at the hip joint early in the support phase during trotting at constant speed on a level surface. Thus, although the forelimb of dogs appears to function as a compliant strut, the hindlimb functions as a lever early in stance phase. Nevertheless, our results indicate that when dogs run at constant speed on a level surface a primary function of both the retractor and protractor muscles of the hindlimb is to produce swing phase of the limb. When the trotting dogs did net work in the fore-aft direction, by running uphill or downhill or by resisting a horizontally directed force, recruitment of the protractor and retractor muscles of the hip joint increased or decreased in the anticipated fashion. These observations are consistent with the hypothesis that recruitment of the oblique hypaxial muscles in trotting dogs function to stabilize the trunk against torques produced by protractor and retractor muscles of the hindlimb. PMID- 19282502 TI - Function of the epaxial muscles during trotting. AB - In mammals, the epaxial muscles are believed to stabilize the trunk during walking and trotting because the timing of their activity is not appropriate to produce bending of the trunk. To test whether this is indeed the case, we recorded the activity of the m. multifidus lumborum and the m. longissimus thoracis et lumborum at three different sites along the trunk (T13, L3, L6) as we manipulated the moments acting on the trunk and the pelvis in dogs trotting on a treadmill. Confirming results of previous studies, both muscles exhibited a biphasic and bilateral activity. The higher burst was associated with the second half of ipsilateral hindlimb stance phase, the smaller burst occurred during the second half of ipsilateral hindlimb swing phase. The asymmetry was noticeably larger in the m. longissimus thoracis et lumborum than in the m. multifidus lumborum. Although our manipulations of the inertia of the trunk produced results that are consistent with previous studies indicating that the epaxial muscles stabilize the trunk against accelerations in the sagittal plane, the responses of the epaxial muscles to manipulations of trunk inertia were small compared with their responses when moments produced by the extrinsic muscles of the hindlimb were manipulated. Our results indicate that the multifidus and longissimus muscles primarily stabilize the pelvis against (1) vertical components of hindlimb retractor muscles and (2) horizontal components of the hindlimb protractor and retractor muscles. Consistent with this, stronger effects of the manipulations were observed in the posterior sampling sites. PMID- 19282503 TI - Eutopic endometrium from women with endometriosis shows altered ultrastructure and glycosylation compared to that from healthy controls--a pilot observational study. AB - Endometrial curettings from a cohort of 24 women with endometriosis were compared with matched biopsies from 14 healthy, fertile women and examined for ultrastructural changes and the secretion of glycans bound by the lectin from Dolichos biflorus. Ultrastructural analysis of glandular endometrial tissue from women with stages I to III endometriosis showed heterogeneous responses to the disease, biopsies often showing a mixture of features, combining delays in the maturation sequence with characteristics of later phenotypes particularly in the mid-late secretory phase of the menstrual cycle. Expression of glycans bound by Dolichos biflorus agglutinin was very variable in these cases but generally matched the observed ultrastructure. Biopsies from women with stage IV endometriosis showed immature gland morphology later in the cycle and also failed to express Dolichos biflorus agglutinin-binding glycans, suggesting an association between histological and biochemical function in advanced disease states. These findings may explain in part endometriosis-associated subfertility as blastocyst attachment is intimately associated with appropriate glycosylation and gland morphology. PMID- 19282504 TI - Life sciences on the Semantic Web: the Neurocommons and beyond. AB - Translational research, the effort to couple the results of basic research to clinical applications, depends on the ability to effectively answer questions using information that spans multiple disciplines. The Semantic Web, with its emphasis on combining information using standard representation languages, access to that information via standard web protocols, and technologies to leverage computation, such as in the form of inference and distributable query, offers a social and technological basis for assembling, integrating and making available biomedical knowledge at Web scale. In this article, we discuss the use of Semantic Web technology for assembling and querying biomedical knowledge from multiple sources and disciplines. We present the Neurocommons prototype knowledge base, a demonstration intended to show the feasibility and benefits of using these technologies. The prototype knowledge base can be used to experiment with and assess the scalability of current tools and methods for creating such a resource, and to elicit issues that will need to be addressed in order to expand the scope and use of it. We demonstrate the utility of the knowledge base by reviewing a few example queries that provide answers to precise questions relevant to the understanding of disease. All components of the knowledge base are freely available at http://neurocommons.org/, enabling readers to reconstruct the knowledge base and experiment with this new technology. PMID- 19282505 TI - The effectiveness of intervention on the behavior of individuals with autism: a meta-analysis using percentage of data points exceeding the median of baseline phase (PEM). AB - The aim of the present study is to demonstrate the percentage of data points exceeding the median of baseline phase (PEM) approach using data on autism treatment for illustrative purposes to compare the effectiveness of different interventions on the problem behaviors of individuals with autism. Electronic databases such as The ProQuest and Google were searched. A total of 163 articles were located, producing 1,502 effect sizes. The results demonstrate that five highly effective intervention strategies were priming, self-control, training, positive reinforcement and punishment, and presenting preferential activities. The least effective strategy was to teach perspective-taking skills. The PEM approach is recommended for use in meta-analysis for single-case experimental designs. PMID- 19282506 TI - Adapted behavior therapy for persistently depressed primary care patients: an open trial. AB - Major depressive disorder is commonly treated in primary care settings. Psychotherapy occurring in primary care should take advantage of the unique aspects of the setting and must adapt to the problems and limitations of the setting. In this open trial, the authors used a treatment development model to adapt behavior therapy for primary care patients (n = 12) with persistent symptoms of depression, despite antidepressant medication treatment. Ten of 12 participants completed 10 sessions of therapy over the course of 4 months, and all endorsed high levels of treatment satisfaction. Participants' depression scores declined significantly over time, and 75% of participants experienced at least 50% change on a self-report measure of depression symptoms. There were trends for social functioning, pain, and general health perceptions to improve over time. These results highlight the acceptability and feasibility of adapting behavior therapy for primary care, and support the continuation of this research. PMID- 19282507 TI - Biomechanical analysis comparing a traditional superior-inferior arthroscopic rotator interval closure with a novel medial-lateral technique in a cadaveric multidirectional instability model. AB - BACKGROUND: Commonly performed arthroscopic rotator interval closure techniques that imbricate the rotator interval in a superior-inferior direction have been unable to reproduce the stabilizing effects of an open medial-lateral rotator interval imbrication. HYPOTHESIS: The medial-lateral rotator interval closure will allow less inferior and posterior glenohumeral translation than the superior inferior rotator interval closure, and the medial-lateral rotator interval closure will result in less loss of external rotation than the superior-inferior closure. STUDY DESIGN: Controlled laboratory study. METHODS: Eight match-paired cadaveric shoulders were stretched to 10% beyond the maximum range of motion in 0 degrees and 60 degrees of glenohumeral abduction to create a multidirectional instability model. Shoulders were then repaired using a superior-inferior rotator interval closure or an arthroscopic medial-lateral rotator interval closure with an anchor in the humeral head. Rotational range of motion, glenohumeral translation, and humeral head apex position were measured for intact, stretched, and repaired conditions in both 0 degrees and 60 degrees of glenohumeral abduction. RESULTS: In 0 degrees of abduction, after both rotator interval closure techniques, external rotation decreased significantly (by 4.4%; P < .05) relative to the stretched state and was restored to the intact state. In 60 degrees of abduction, only the medial-lateral rotator interval closure restored range of motion to the intact state. In 60 degrees of abduction, the medial lateral rotator interval closure was more effective in reducing posterior translation than was the superior-inferior closure (P = .03). CONCLUSION: The medial-lateral rotator interval closure restored range of motion to the intact state better than the superior-inferior closure. Compared with the superior inferior rotator interval closure, the medial-lateral closure significantly decreased posterior translation with the shoulder in abduction and external rotation. CLINICAL RELEVANCE: Arthroscopic medial-lateral rotator interval closure with a suture anchor in the humeral head can be considered in the surgical treatment of patients with multidirectional instability, especially those with a component of posterior instability, without concern for excessive loss of range of motion. PMID- 19282508 TI - Delayed vastus medialis obliquus to vastus lateralis onset timing contributes to the development of patellofemoral pain in previously healthy men: a prospective study. AB - BACKGROUND: Delayed onset of vastus medialis obliquus activity has been described in patellofemoral pain patients. No prospective study investigating the development of patellofemoral pain has tested the onset timing of electromyographic activity of the vastus medialis obliquus and vastus lateralis muscles during a functional task. HYPOTHESIS: Before the development of patellofemoral pain, subjects demonstrate an altered firing order of the vastus medialis obliquus and vastus lateralis muscles compared with healthy subjects during a functional task. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: The onset of vastus medialis obliquus and vastus lateralis activity was measured with surface electromyography during a functional task (rocking back on the heels) in 79 healthy subjects subsequently submitted to a 6-week strenuous basic military training. Afterward, these subjects were reassessed. RESULTS: Thirty-two percent of the recruits developed patellofemoral pain during training. Patellofemoral pain subjects demonstrated a significant delay of onset of vastus medialis obliquus electromyographic activity compared with the healthy controls (P = .023), even before basic military training. In healthy subjects, no significant differences in electromyographic activity onset of the vastus medialis obliquus compared with the vastus lateralis could be identified before and after basic military training. A significant (P < .001) delay could be demonstrated in the patellofemoral pain group after basic military training. A binary logistic regression could be constructed wherein the onset of the electromyographic activity of the vastus medialis obliquus and vastus lateralis was withheld in the model. The most optimal cutoff value, which is based on the receiver operating characteristic curve, is a timing difference of -0.67 milliseconds (vastus medialis obliquus - vastus lateralis). The area under the receiver operating characteristic curve is considered as fair (0.68). CONCLUSION: Delayed onset of electromyographic activity of the vastus medialis obliquus vastus lateralis is one of the contributing risk factors to the development of patellofemoral pain. PMID- 19282509 TI - Use of autologous platelet-rich plasma to treat muscle strain injuries. AB - BACKGROUND: Standard nonoperative therapy for acute muscle strains usually involves short-term rest, ice, and nonsteroidal anti-inflammatory medications, but there is no clear consensus on how to accelerate recovery. HYPOTHESIS: Local delivery of platelet-rich plasma to injured muscles hastens recovery of function. STUDY DESIGN: Controlled laboratory study. METHODS: In vivo, the tibialis anterior muscles of anesthetized Sprague-Dawley rats were injured by a single (large strain) lengthening contraction or multiple (small strain) lengthening contractions, both of which resulted in a significant injury. The tibialis anterior either was injected with platelet-rich plasma, was injected with platelet-poor plasma as a sham treatment, or received no treatment. RESULTS: Both injury protocols yielded a similar loss of force. The platelet-rich plasma only had a beneficial effect at 1 time point after the single contraction injury protocol. However, platelet-rich plasma had a beneficial effect at 2 time points after the multiple contraction injury protocol and resulted in a faster recovery time to full contractile function. The sham injections had no effect compared with no treatment. CONCLUSION: Local delivery of platelet-rich plasma can shorten recovery time after a muscle strain injury in a small-animal model. Recovery of muscle from the high-repetition protocol has already been shown to require myogenesis, whereas recovery from a single strain does not. This difference in mechanism of recovery may explain why platelet-rich plasma was more effective in the high-repetition protocol, because platelet-rich plasma is rich in growth factors that can stimulate myogenesis. CLINICAL RELEVANCE: Because autologous blood products are safe, platelet-rich plasma may be a useful product in clinical treatment of muscle injuries. PMID- 19282510 TI - Gene expression profile of rat ovarian tissue following xenotransplantation into immune-deficient mice. AB - Immune-compromised mice have been used as gonadal tissue recipients to develop gametes of various mammalian species. The aim of this research was to determine gene expression differences between fresh and frozen-thawed rat xenotransplanted (XT) ovaries as well the gene expression differences between XT and sexually mature rat ovaries that were non-transplanted (NT). Ovaries from sexually immature female rats were transplanted under the kidney capsule of ovariectomized athymic nude mice either fresh or after freezing. The XT ovaries were collected approximately 10-12 weeks after xenografting for microarray analysis. The NT ovaries were collected from sexually mature rats. Gene expression was very similar between fresh and cryopreserved XT ovaries: 125 genes were twofold up- or downregulated, but level of regulation was not statistically significant. Overall patterns of gene expression between XT and NT ovaries were very different indicated by the absence of diagonal relationship between XT and NT ovary gene expression. More than 3000 genes were significantly (P<0.01) up- or downregulated between XT and NT ovaries. Genes involved in metabolic processes, lipid metabolism, and growth were downregulated in XT ovaries, whereas genes involved in immune and inflammatory response were upregulated in XT ovaries. The results showed that ovarian tissue xenografting significantly alters genes responsible for ovarian metabolism and function and leads to an upregulation of genes responsible for graft rejection. PMID- 19282511 TI - SOD-1 deletions in Caenorhabditis elegans alter the localization of intracellular reactive oxygen species and show molecular compensation. AB - Superoxide dismutase (SOD) is an enzyme that catalytically removes the superoxide radical (*O2-) and protects organisms from oxidative damage during normal aging. We demonstrate that not only the cytosolic *O2- level but also the mitochondrial *O2- level increases in the deletion mutants of sod-1 gene encoding Cu/Zn SOD in Caenorhabditis elegans (C. elegans). Interestingly, this suggests that the activity of SOD-1, which so far has been thought to act mainly in cytoplasm, helps to control the detoxification of *O2- also in the mitochondria. We also found functional compensation by other SODs, especially the sod-5 gene, which was induced several fold in the mutants. Therefore, the possibility exists that the compensative expression of sod-5 gene in the sod-1 deficit is associated with the insulin/insulin-like growth factor-1 (Ins/IGF-1) signaling pathway, which regulates longevity and stress resistance of C. elegans because the sod-5 gene may be a target of the pathway. PMID- 19282512 TI - The NBPF1 promoter has been recruited from the unrelated EVI5 gene before simian radiation. AB - Most new genes arise through the duplication of existing genes. In most cases, the duplication is not limited to the coding sequence but encompasses the regulatory region as well. The NBPF gene family has expanded during recent primate evolution, and it has no known mouse ortholog. One of its members, NBPF1, was found to be disrupted by a constitutional translocation in a neuroblastoma patient. Here, we show that the ancestral NBPF gene copied the regulatory region from an unrelated gene, EVI5, after the split between simians and prosimians but before simian radiation. Phylogenetic analysis points to the possible involvement of positive selection acting on the NBPF1 promoter in the simian lineage. We previously showed decreased NBPF1 expression in certain neuroblastoma cell lines. Here, we show that this expression pattern is mimicked by the EVI5 gene, but partly by different mechanisms. Epigenetic regulation of the EVI5 promoter is common in neuroblastoma cell lines, but it is not for the NBPF promoters. Here, we describe the recent acquisition of the NBPF1 promoter from an unrelated gene, and remarkably, both the donor (EVI5) and acceptor (NBPF1) genes are disrupted by constitutional translocations in patients with neuroblastoma, suggesting a functional link between these genes and the disease. PMID- 19282513 TI - Differences in DNA methylation patterns and expression of the CCRK gene in human and nonhuman primate cortices. AB - Changes in DNA methylation patterns during embryo development and differentiation processes are linked to the transcriptional plasticity of our genome. However, little is known about the evolutionary conservation of DNA methylation patterns and the evolutionary impact of epigenetic differences between closely related species. Here we compared the methylation patterns of CpG islands (CGIs) in the promoter regions of seven genes in humans and chimpanzees. We identified a block of CpGs in the cell cycle-related kinase (CCRK) gene that is more methylated in the adult human cortex than in the chimpanzee cortex and, in addition, it exhibits considerable intraspecific variation both in humans and chimpanzees. The species-specifically methylated region (SMR) lies between the almost completely methylated 5' region and the completely demethylated 3' region of the presumed CCRK CGI promoter. It is part of an Alu-Sg1 repeat that has been integrated into the promoter region in a common ancestor of humans and New World monkeys. This SMR is relatively hypomethylated in the rhesus monkey cortex and more or less completely methylated in the baboon cortex, indicating extraordinary methylation dynamics during primate evolution. The mRNA expression level of CCRK has also changed during the course of primate evolution. CCRK is expressed at much higher levels in human and baboon cortices, which display an average SMR methylation of 70% and 100%, respectively, than in chimpanzee and rhesus macaque cortices with an average SMR methylation of 35% and 40%, respectively. The observed evolutionary dynamics suggests a possibility that CCRK has been important for evolution of the primate brain. PMID- 19282514 TI - The substance, scholarship, and science of research on violence against women: a comment. PMID- 19282515 TI - Should the study of violence against women be a science? PMID- 19282516 TI - Advancing the study of violence against women: response to Jordan. PMID- 19282517 TI - Science, social change, and ending violence against women: which one of these is not like the others? PMID- 19282518 TI - Advancing the study of violence against women: response to commentaries and next steps. PMID- 19282519 TI - Intimate partner violence against Athabaskan women residing in interior Alaska: results of a victimization survey. AB - A survey instrument mirroring the National Violence Against Women Survey was administered in person to measure the incidence and prevalence of intimate partner violence against Athabaskan women residing in the interior of Alaska. Roughly two thirds of respondents (63.7%) reported an intimate partner assault victimization at some point in their adult lifetime, and 18% of the respondents reported that they had been physically assaulted by an intimate partner in the year preceding the survey. Findings revealed that intimate partner assault victimization is more prevalent and is considerably more frequent when compared to that reported for American women in general. PMID- 19282522 TI - In defence of a "good-enough" family physician. PMID- 19282524 TI - To be good enough. PMID- 19282526 TI - Sleep apnea and gout. PMID- 19282527 TI - Treating chronic pain. PMID- 19282528 TI - New tool for FPs. PMID- 19282529 TI - Government billing interferes with standards of care. PMID- 19282530 TI - Approach to management of suspected rabies exposures: what primary care physicians need to know. AB - OBJECTIVE: To review the role of primary care physicians, in conjunction with local public health units, in the management of suspected rabies exposures and to outline the current guidelines for the administration of rabies postexposure prophylaxis. SOURCES OF INFORMATION: Published guidelines on the topic of rabies were reviewed and additional articles were identified from key references. Various public health websites were also explored. Most evidence was level II or III. MAIN MESSAGE: Primary care physicians must always consider the risk of rabies when treating patients who have had animal-to-human exposures (eg, bite, scratch), and if indicated, postexposure prophylaxis must be administered as soon as possible because the infection is fatal once clinical symptoms develop. CONCLUSION: Human cases of rabies are almost entirely preventable if suspected exposures are identified and managed promptly and properly. Primary care physicians must continue to work together with local public health officials in order to minimize the threat of this deadly virus. PMID- 19282531 TI - Vaginal yeast infections during pregnancy. AB - QUESTION: My pregnant patients often present with symptomatic vaginal yeast infections. Are the medications commonly used for the management of yeast infections safe to use during pregnancy? ANSWER: Existing data indicate that exposure to oral and topical antifungals, topical antiseptics, or corticosteroids during pregnancy is not associated with increased risk of major malformations. Topical azole antifungals are the recommended treatment during pregnancy for at least 7 days owing to increased efficacy. Topical corticosteroids can be used for symptomatic relief. PMID- 19282532 TI - Should weight-loss supplements be used for pediatric obesity? AB - QUESTION: In my clinic I have a large population of overweight and obese children. There is a range of weight-loss supplements marketed for the adult population. What natural health products for treatment of obesity are effective and can be used in children? ANSWER: Weight-loss supplements lack sufficient data supporting their efficacy and safety, even in adults. Most weight-loss supplements cannot be recommended at this time for children. Options for obese adolescents include increasing consumption of fibre with diet or using fibre supplements, such as glucomannan. Dietary fibres can also prevent side effects of orlistat, the only medication available for treatment of obese adolescents. PMID- 19282533 TI - Can't we get this over with?: an approach to assessing the patient who requests hastened death. PMID- 19282534 TI - Maxillary carcinoma: a wolf in sheep's clothing. PMID- 19282536 TI - Taking the stress out of acne management. PMID- 19282537 TI - Dermacase. Auricular pseudocyst. PMID- 19282538 TI - Bridging the gap between primary care and the cancer system: the UPCON Network of CancerCare Manitoba. AB - PROBLEM BEING ADDRESSED: Patient care is poorly coordinated between family physicians and the cancer system and the working relationships are not strong. OBJECTIVE OF PROGRAM: To improve integration of patient care and communication between FPs and cancer specialists; enhance FPs' knowledge of cancer and the cancer system; and promote the role of primary care within the cancer care system. PROGRAM DESCRIPTION: The Uniting Primary Care and Oncology (UPCON) Network of CancerCare Manitoba has created partnerships with 12 primary care clinics in Winnipeg, Man, by providing the following: access to the provincial electronic medical record for cancer; small group continuing professional development for a "lead physician" from each clinic to make him or her the local cancer resource; educational outreach to all clinic staff; and changes within CancerCare Manitoba to highlight the role of FPs. CONCLUSION: Lead physicians are appreciated by their clinic colleagues, and these FPs are the main users of the cancer electronic medical record. A strong cancer continuing professional development program has been implemented and a voice for primary care has been created within the agency. The UPCON Network is now expanding throughout Manitoba. PMID- 19282539 TI - Disrespect, harassment, and abuse: all in a day's work for family physicians. AB - OBJECTIVE: To examine harassment and abusive encounters between family physicians and their patients or colleagues in the workplace. DESIGN: Qualitative case study using semistructured interviews. SETTING: Province of New Brunswick. PARTICIPANTS: Forty-eight family physicians from across the province. METHODS: A collective case-study approach was developed, with 24 cases of 2 individuals per case. Cases were selected based on sex, location (urban or rural), language (French or English), and number of years since medical school graduation (< 10 years, 10 to 20 years, or > 20 years). Physicians were interviewed in either French or English. Participants were recruited using the College of Physicians and Surgeons of New Brunswick's physician directory. Based on the rates of response and participation, some cases were overrepresented, while others were not completed. All interviews were audiotaped, transcribed verbatim, and analyzed thematically using a categorical aggregation approach. A coding scheme for the thematic analysis was developed by the research team before the interviews were transcribed. MAIN FINDINGS: Although the original intent of this study was to examine the work environment of family physicians in light of the increasing number of women entering the profession, harassment and abusive encounters in the workplace emerged as a main theme. These encounters ranged from minor to severe. Minor abusive encounters included disrespectful behaviour and verbal threats by patients, their families, and occasionally colleagues. More severe forms of harassment involved physical threats, physical encounters, and stalking. Demanding patients, such as heavy drug users, were often seen as threatening. Location of practice, years in practice, and sex of the physician seemed to affect abusive encounters--young, female, rural physicians appeared to experience such encounters most often. CONCLUSION: Abusive encounters in the workplace are concerning. It is essential to address these issues of workplace harassment and abuse in order to protect physician safety and avoid workplace dissatisfaction. Abusive encounters might push family physicians to leave clinical practice prematurely or refuse to work in higher-risk environments, such as emergency departments or rural areas. PMID- 19282540 TI - Crossing boundaries: family physicians' struggles to protect their private lives. AB - OBJECTIVE: To explore the tensions between professional and personal boundaries and how they affect the work and private lives of family physicians. DESIGN: Qualitative case study using semistructured interviews. SETTING: Province of New Brunswick. PARTICIPANTS: Forty-eight family physicians from across the province. METHODS: A collective case-study approach was developed, with 24 cases of 2 individuals per case. Cases were selected based on sex, location (urban or rural), language (French or English), and number of years since medical school graduation (< 10 years, 10 to 20 years, or > 20 years). Physicians were interviewed in either French or English. Participants were recruited using the College of Physicians and Surgeons of New Brunswick's physician directory. Based on the rates of response and participation, some cases were overrepresented, while others were not completed. All interviews were audiotaped, transcribed verbatim, and analyzed thematically using a categorical aggregation approach. A coding scheme for the thematic analysis was developed by the research team before the interviews were transcribed. MAIN FINDINGS: Almost all of the family physicians interviewed discussed how their profession negatively affected their personal lives. Many struggled with issues such as heavy workloads, the adverse effects of their profession on their family lives, and the trespassing of patients onto their personal lives in small towns and rural communities. Some physicians had developed strategies to balance their personal lives with their professional demands; however, this often meant reducing work hours or terminating certain shifts, such as those in the emergency department or after hours clinics. CONCLUSION: Family physicians struggle to keep their profession from intruding too much into their private lives. These struggles are important to acknowledge and address in order to avoid physician burnout and premature retirement from clinical practice. PMID- 19282541 TI - Exploring family physician stress: helpful strategies. AB - OBJECTIVE: To explore the nature of professional stress and the strategies used by family physicians to deal with this stress. DESIGN: Qualitative study. SETTING: Kitchener-Waterloo, Ont. PARTICIPANTS: Ten key-informant family physicians. METHODS: In-depth interviews were conducted with key informants. A total of 40 key informants were identified, based on selected criteria; 24 provided consent. The potential participants were rank-ordered for interviews to provide maximum variation in age, sex, and years in practice. Interviews were conducted, audiotaped, transcribed verbatim, and analyzed until thematic saturation was reached, as determined through an iterative process. This occurred after 10 in-depth interviews. Immersion and crystallization techniques were used. MAIN FINDINGS: The participants described professional stresses and strategies at the personal, occupational, and health care system levels. Personal stressors included personality traits and the need to balance family and career, which were countered by biological, psychological, social, and spiritual strategies. Occupational stressors included challenging patients, high workload, time limitations, competency issues, challenges of documentation and practice management, and changing roles within the workplace. Occupational stressors were countered by strategies such as setting limits, participating in continuing medical education, soliciting support from colleagues and staff, making use of teams, improving patient-physician relationships, exploring new forms of remuneration, and scheduling appropriately. Stressors affecting the wider health care system included limited resources, imposed rules and regulations, lack of support from specialists, feeling undervalued, and financial concerns. CONCLUSION: Family physicians face a multitude of challenges at personal, occupational, and health care system levels. A systems approach provides a new framework in which proactive strategies can augment more than one level of a system and, in contrast, reactive strategies can have negative inputs for different system levels. PMID- 19282544 TI - Multithreaded hybrid feature tracking for markerless augmented reality. AB - We describe a novel markerless camera tracking approach and user interaction methodology for augmented reality (AR) on unprepared tabletop environments. We propose a real-time system architecture that combines two types of feature tracking. Distinctive image features of the scene are detected and tracked frame to-frame by computing optical flow. In order to achieve real-time performance, multiple operations are processed in a synchronized multi-threaded manner: capturing a video frame, tracking features using optical flow, detecting distinctive invariant features, and rendering an output frame. We also introduce user interaction methodology for establishing a global coordinate system and for placing virtual objects in the AR environment by tracking a user's outstretched hand and estimating a camera pose relative to it. We evaluate the speed and accuracy of our hybrid feature tracking approach, and demonstrate a proof-of concept application for enabling AR in unprepared tabletop environments, using bare hands for interaction. PMID- 19282545 TI - Mixed reality humans: evaluating behavior, usability, and acceptability. AB - This paper presents Mixed Reality Humans (MRHs), a new type of embodied agent enabling touch-driven communication. Affording touch between human and agent allows MRHs to simulate interpersonal scenarios in which touch is crucial. Two studies provide initial evaluation of user behavior with a MRH patient and the usability and acceptability of a MRH patient for practice and evaluation of medical students' clinical skills. In Study I (n=8) it was observed that students treated MRHs as social actors more than students in prior interactions with virtual human patients (n=27), and used interpersonal touch to comfort and reassure the MRH patient similarly to prior interactions with human patients (n=76). In the within-subjects Study II (n=11), medical students performed a clinical breast exam on each of a MRH and human patient. Participants performed equivalent exams with the MRH and human patients, demonstrating the usability of MRHs to evaluate students' exam skills. The acceptability of the MRH patient for practicing exam skills was high as students rated the experience as believable and educationally beneficial. Acceptability was improved from Study I to Study II due to an increase in the MRH's visual realism, demonstrating that visual realism is critical for simulation of specific interpersonal scenarios. PMID- 19282546 TI - Evaluation of reorientation techniques and distractors for walking in large virtual environments. AB - Virtual Environments (VEs) that use a real-walking locomotion interface have typically been restricted in size to the area of the tracked lab space. Techniques proposed to lift this size constraint, enabling real walking in VEs that are larger than the tracked lab space, all require reorientation techniques (ROTs) in the worst-case situation-when a user is close to walking out of the tracked space. We propose a new ROT using visual and audial distractors-objects in the VE that the user focuses on while the VE rotates-and compare our method to current ROTs through three user studies. ROTs using distractors were preferred and ranked more natural by users. Our findings also suggest that improving visual realism and adding sound increased a user's feeling of presence. Users were also less aware of the rotating VE when ROTs with distractors were used. Our findings also suggest that improving visual realism and adding sound increased a user's feeling of presence. PMID- 19282547 TI - Semiautomatic transfer function initialization for abdominal visualization using self-generating hierarchical radial basis function networks. AB - As being a tool that assigns optical parameters used in interactive visualization, Transfer Functions (TF) have important effects on the quality of volume rendered medical images. Unfortunately, finding accurate TFs is a tedious and time consuming task because of the trade off between using extensive search spaces and fulfilling the physician's expectations with interactive data exploration tools and interfaces. By addressing this problem, we introduce a semi automatic method for initial generation of TFs. The proposed method uses a Self Generating Hierarchical Radial Basis Function Network to determine the lobes of a Volume Histogram Stack (VHS) which is introduced as a new domain by aligning the histograms of slices of a image series. The new self generating hierarchical design strategy allows the recognition of suppressed lobes corresponding to suppressed tissues and representation of the overlapping regions which are parts of the lobes but can not be represented by the Gaussian bases in VHS. Moreover, approximation with a minimum set of basis functions provides the possibility of selecting and adjusting suitable units to optimize the TF. Applications on different CT and MR data sets show enhanced rendering quality and reduced optimization time in abdominal studies. PMID- 19282548 TI - A novel visualization technique for electric power grid analytics. AB - The application of information visualization holds tremendous promise for the electric power industry, but its potential has so far not been sufficiently exploited by the visualization community. Prior work on visualizing electric power systems has been limited to depicting raw or processed information on top of a geographic layout. Little effort has been devoted to visualizing the physics of the power grids, which ultimately determines the condition and stability of the electricity infrastructure. Based on this assessment, we developed a novel visualization system prototype, GreenGrid, to explore the planning and monitoring of the North American Electricity Infrastructure. The paper discusses the rationale underlying the GreenGrid design, describes its implementation and performance details, and assesses its strengths and weaknesses against the current geographic-based power grid visualization. We also present a case study using GreenGrid to analyze the information collected moments before the last major electric blackout in the Western United States and Canada, and a usability study to evaluate the practical significance of our design in simulated real-life situations. Our result indicates that many of the disturbance characteristics can be readily identified with the proper form of visualization. PMID- 19282549 TI - Visualization of simulated urban spaces: inferring parameterized generation of streets, parcels, and aerial imagery. AB - Urban simulation models and their visualization are used to help regional planning agencies evaluate alternative transportation investments, land use regulations, and environmental protection policies. Typical urban simulations provide spatially distributed data about number of inhabitants, land prices, traffic, and other variables. In this article, we build on a synergy of urban simulation, urban visualization, and computer graphics to automatically infer an urban layout for any time step of the simulation sequence. In addition to standard visualization tools, our method gathers data of the original street network, parcels, and aerial imagery and uses the available simulation results to infer changes to the original urban layout and produce a new and plausible layout for the simulation results. In contrast with previous work, our approach automatically updates the layout based on changes in the simulation data and thus can scale to a large simulation over many years. The method in this article offers a substantial step forward in building integrated visualization and behavioral simulation systems for use in community visioning, planning, and policy analysis. We demonstrate our method on several real cases using a 200 GB database for a 16,300 km2 area surrounding Seattle. PMID- 19282550 TI - Equalizer: a scalable parallel rendering framework. AB - Continuing improvements in CPU and GPU performances as well as increasing multi core processor and cluster-based parallelism demand for flexible and scalable parallel rendering solutions that can exploit multipipe hardware accelerated graphics. In fact, to achieve interactive visualization, scalable rendering systems are essential to cope with the rapid growth of data sets. However, parallel rendering systems are non-trivial to develop and often only application specific implementations have been proposed. The task of developing a scalable parallel rendering framework is even more difficult if it should be generic to support various types of data and visualization applications, and at the same time work efficiently on a cluster with distributed graphics cards. In this paper we introduce a novel system called Equalizer, a toolkit for scalable parallel rendering based on OpenGL which provides an application programming interface (API) to develop scalable graphics applications for a wide range of systems ranging from large distributed visualization clusters and multi-processor multipipe graphics systems to single-processor single-pipe desktop machines. We describe the system architecture, the basic API, discuss its advantages over previous approaches, present example configurations and usage scenarios as well as scalability results. PMID- 19282551 TI - Real-time depth-of-field rendering using anisotropically filtered mipmap interpolation. AB - This article presents a real-time GPU-based post-filtering method for rendering acceptable depth-of-field effects suited for virtual reality. Blurring is achieved by nonlinearly interpolating mipmap images generated from a pinhole image. Major artifacts common in the post-filtering techniques such as bilinear magnification artifact, intensity leakage, and blurring discontinuity are practically eliminated via magnification with a circular filter, anisotropic mipmapping, and smoothing of blurring degrees. The whole framework is accelerated using GPU programs for constant and scalable real-time performance required for virtual reality. We also compare our method to recent GPU-based methods in terms of image quality and rendering performance. PMID- 19282552 TI - An adaptive correspondence algorithm for modeling scenes with strong interreflections. AB - Modeling real-world scenes, beyond diffuse objects, plays an important role in computer graphics, virtual reality, and other commercial applications. One active approach is projecting binary patterns in order to obtain correspondence and reconstruct a densely sampled 3D model. In such structured-light systems, determining whether a pixel is directly illuminated by the projector is essential to decoding the patterns. When a scene has abundant indirect light, this process is especially difficult. In this paper, we present a robust pixel classification algorithm for this purpose. Our method correctly establishes the lower and upper bounds of the possible intensity values of an illuminated pixel and of a non illuminated pixel. Based on the two intervals, our method classifies a pixel by determining whether its intensity is within one interval but not in the other. Our method performs better than standard method due to the fact that it avoids gross errors during decoding process caused by strong inter-reflections. For the remaining uncertain pixels, we apply an iterative algorithm to reduce the inter reflection within the scene. Thus, more points can be decoded and reconstructed after each iteration. Moreover, the iterative algorithm is carried out in an adaptive fashion for fast convergence. PMID- 19282553 TI - Indexing and retrieving motions of characters in close contact. AB - Human motion indexing and retrieval are important for animators due to the need to search for motions in the database which can be blended and concatenated. Most of the previous researches of human motion indexing and retrieval compute the Euclidean distance of joint angles or joint positions. Such approaches are difficult to apply for cases in which multiple characters are closely interacting with each other, as the relationships of the characters are not encoded in the representation. In this research, we propose a topology-based approach to index the motions of two human characters in close contact. We compute and encode how the two bodies are tangled based on the concept of rational tangles. The encoded relationships, which we define as TangleList, are used to determine the similarity of the pairs of postures. Using our method, we can index and retrieve motions such as one person piggy-backing another, one person assisting another in walking, and two persons dancing / wrestling. Our method is useful to manage a motion database of multiple characters. We can also produce motion graph structures of two characters closely interacting with each other by interpolating and concatenating topologically similar postures and motion clips, which are applicable to 3D computer games and computer animation. PMID- 19282554 TI - Direct forcing for Lagrangian rigid-fluid coupling. AB - We propose a novel boundary handling algorithm for particle-based fluids. Based on a predictor-corrector scheme for both velocity and position, one- and two-way coupling with rigid bodies can be realized. The proposed algorithm offers significant improvements over existing penalty-based approaches. Different slip conditions can be realized and non-penetration is enforced. Direct forcing is employed to meet the desired boundary conditions and to ensure valid states after each simulation step. We have performed various experiments in 2D and 3D. They illustrate one- and two-way coupling of rigid bodies and fluids, the effects of hydrostatic and dynamic forces on a rigid body as well as different slip conditions. Numerical experiments and performance measurements are provided. PMID- 19282555 TI - Computing Teichmuller shape space. AB - Shape indexing, classification, and retrieval are fundamental problems in computer graphics. This work introduces a novel method for surface indexing and classification based on Teichmuller theory. The Teichmuller space for surfaces with the same topology is a finite dimensional manifold, where each point represents a conformal equivalence class, a curve represents a deformation process from one class to the other. We apply Teichmuller space coordinates as shape descriptors, which are succinct, discriminating and intrinsic; invariant under the rigid motions and scalings, insensitive to resolutions. Furthermore, the method has solid theoretic foundation, and the computation of Teichmuller coordinates is practical, stable and efficient. This work focuses on the surfaces with negative Euler numbers, which have a unique conformal Riemannian metric with -1 Gaussian curvature. The coordinates which we will compute are the lengths of a special set of geodesics under this special metric. The metric can be obtained by the curvature flow algorithm, the geodesics can be calculated using algebraic topological method. We tested our method extensively for indexing and comparison of about one hundred of surfaces with various topologies, geometries and resolutions. The experimental results show the efficacy and efficiency of the length coordinate of the Teichmuller space. PMID- 19282556 TI - Quasi-developable mesh surface interpolation via mesh deformation. AB - We present a new algorithm for finding a most "developable" smooth mesh surface to interpolate a given set of arbitrary points or space curves. Inspired by the recent progress in mesh editing that employs the concepts of preserving the Laplacian coordinates and handle-based shape editing, we formulate the interpolation problem as a mesh deformation process that transforms an initial developable mesh surface, such as a planar figure, to a final mesh surface that interpolates the given points and/or curves. During the deformation, the developability of the intermediate mesh is maintained by means of preserving the zero-valued Gaussian curvature on the mesh. To treat the high nonlinearity of the geometric constrains owing to the preservation of Gaussian curvature, we linearize those nonlinear constraints using Taylor expansion and eventually construct a sparse and over-determined linear system which is subsequently solved by a robust least-squares solution. By iteratively performing this procedure, the initial mesh is gradually and smoothly "dragged" to the given points and/or curves. The initial experimental data has shown some promising aspects of the proposed algorithm as a general quasi-developable surface interpolation tool. PMID- 19282557 TI - Non-invasive estimation of pulsatile flow and differential pressure in an implantable rotary blood pump for heart failure patients. AB - We propose dynamical models for pulsatile flow and head estimation in an implantable rotary blood pump. Pulsatile flow and head data were obtained using a circulatory mock loop where fluid solutions with different values of viscosities were used as a blood analogue with varying haematocrit (HCT). Noninvasive measurements of power and pump speed were used with HCT values as inputs to the flow model while the estimated flow was used with the speed as inputs to a head estimation model. Linear regression analysis between estimated and measured flows obtained from a mock loop resulted in a highly significant correlation (R2=0.982) and a mean absolute error (e) of 0.323 L min(-1), while for head, R2=0.933 and e=7.682 mmHg were obtained. R2=0.849 and e=0.584 L min(-1) were obtained when the same model derived in the mock loop was used for flow estimation in ex vivo porcine data (N=6). Furthermore, in the steady state, the solution of the presented flow model can be described by a previously designed and verified static model. The models developed herein will play a vital role in developing a robust control system of the pump flow coping with changing physiological demands. PMID- 19282558 TI - Electrical and mechanical effects of hyoscine butylbromide on the human stomach: a non-invasive approach. AB - Due to the importance of motility in a number of gastrointestinal disorders, efforts have been made to evaluate both gastric motility counterparts: electrical activity and mechanical activity. The present work aimed to propose a new approach, associating AC biosusceptometry (ACB) and electrogastrography (EGG), to noninvasively monitoring mechanical and electrical gastric activity, respectively. Fourteen volunteers ingested a test meal and their gastric activity was evaluated by EGG and ACB at a baseline and after 20 mg of i.v. hyoscine butylbromide. ACB and EGG showed a similar signal pattern and high temporal correlation. Hyoscine butylbromide decreased the mechanical motility index (MI) by 50.9%, while for electrical MI the reduction was 36.5%. Delayed times to onset (mean+/-SD: 50+/-15 versus 40+/-20 s; P<0.01) and the inhibitory effect (16+/-4 versus 14+/-5 min; P<0.01) were calculated for ACB and EGG, respectively. ACB and EGG emerged due to their interesting nature, noninvasiveness and low cost to evaluate gastric motility. Our approach associating ACB and EGG allowed monitoring and quantification of the effects of an anticholinergic drug in gastric electrical activity and contractile activity in humans. PMID- 19282559 TI - Tackling the cancer Tsunami. PMID- 19282560 TI - Cell phones and tumor: still in no man's land. AB - The use of cell phones is increasing worldwide at a phenomenal pace. While cellular communication has dramatically influenced our lifestyle, its impact on human health has not been completely assessed. Widespread concern continues in the community about the deleterious effects of radiofrequency radiations (with which cell phones operate) on human tissues and the subsequent potential for carcinogenesis. A detailed survey of published studies researching this question was done in preparation of this manuscript. Included in the survey were case reports, in vitro studies, population based retrospective studies and other investigations. The database of indexed journals was searched for key words like 'cell phone', 'radiation', 'cancer' and 'radio waves'. Guidelines issued by the World Health Organization, federal and technical authorities, Institute of Electrical and Electronic Engineers and the International Commission for Non Ionizing Radiation Protection were reviewed. The evaluation of current evidence provided by various studies to suggest the possible carcinogenic potential of radiofrequency radiation is inconclusive. This risk assumes significance in light of the burgeoning number of people who are continually exposed to the high frequency radiation from cell phones and towers that serve as receiving and transmitting stations. The aim of this review is to identify limitations in past studies, present available data for consideration, and identify gaps in the current knowledge base. This will provide impetus and direction for further research and allow informed decisions pertaining to cell phone use to be made. PMID- 19282561 TI - Oncologists should measure life, not death: a newer perspective of cancer statistics. AB - We discuss why we should measure parameters of 'life' and not 'death' in choosing a therapy for cancer and what are the flaws and difficulties in measuring cancer specific deaths. We then submit those parameters which can be measured with high precision and validity in cancer patients and so should be recorded to guide us in deciding the best therapy. PMID- 19282562 TI - Endocrinological cancers and the internet. AB - Endocrine cancers are some of the commonest forms of cancer getting mention as early as in old papyruses of Egypt. Our current knowledge about this cluster of cancer has grown immensely with improved survival rates. In the era of specialization and super specialization, we started peeking into these conditions more elaborately and beyond microscopically. The growing awareness demanded categorization of information into disease basics like molecular pathology of initiation and progression; advanced diagnostics; new therapeutic options; patient awareness and involvement in clinical trials. Recent advances in genomics and hereditary counseling have delineated pre-disease forecasting possibilities. With advanced diagnostics and therapeutic modalities, we saw an increase in cancer survivors demanding extra care and moral support. Throughout these developments, we went through a boost in global information communications, the main thrust being the Internet. Networking of computers globally generated a platform that created a ripple of knowledge far and wide. The purpose of this review is to investigate how the Internet is supporting the growth and development of the field of endocrine cancer, and present and future scope of the Internet as a tool for professionals involved in this area. The information furnished here were collected from cited references as well as all websites mentioned in the tables. PMID- 19282563 TI - Study of 'patterns of care' of ovarian cancer patients in a specialized cancer institute in Kolkata, eastern India. AB - BACKGROUND: Ovarian cancer is one of the leading cancers in Indian women. The current standard of care is a combination of surgical staging, maximal surgical effort to achieve cytoreduction, and judicious use of chemotherapy. Multimodality therapy can reduce mortality, but the practice and uptake of such therapy in Indian Institutions are not up to the desired level. OBJECTIVES: To study the protocol adherence for ovarian cancer management along with patient compliance and evaluate their effects on survival. MATERIALS AND METHODS: The retrospective study obtained and analyzed data from records of patients operated for ovarian cancer at a Regional Cancer Center in eastern India between January 2002 and December 2006. RESULTS: The records of 202 patients were evaluable. None of the patients who had primary surgery outside the institute had staging information. A substantial number of patients operated at the institute had incomplete surgical staging, inadequate information on residual disease after surgery, and incomplete histology report. Only 20.3% patients could have optimal cytoreduction after surgery. Compliance to chemotherapy was poor. The median overall survival time and disease-free survival time were 24 months and 5 months, respectively. The residual disease after surgery significantly affected the overall survival, but not the disease-free survival. Incomplete chemotherapy was found to adversely affect survival after adjusting for advanced stage and bulky residual disease. CONCLUSION: Management of ovarian cancer is suboptimal even in the specialized cancer institute. Poor patient compliance to chemotherapy is one of the major factors adversely affecting survival from advanced ovarian cancer. PMID- 19282564 TI - Desmoid tumors: experience of 32 cases and review of the literature. AB - BACKGROUND: Desmoids are infiltrative, locally destructive, soft tissue tumors. Although they do not metastasize, the incidence of local recurrence is quite high. AIM: Present study aimed at reporting the 10-year experience of 32 desmoid cases and reviewing some facts with symptoms, investigation, and treatment of the disease. MATERIALS AND METHODS: Thirty two cases of desmoid tumors were reviewed over a 10-year span. Surgical resection and adjuvant radiotherapy were the treatments of choice whenever histological margins were positive. RESULTS: Multiparous women in reproductive age were the most commonly afflicted. The commonest site of presentation was the abdominal wall. Ninety one percent (20/22) were infraumbilical. The tumors were found in the rectus sheath in 14 patients (64%) and were laterally situated in 8 patients (36%). Local infiltration was found in six patients and the urinary bladder was most commonly involved (3/6). Locally recurrent desmoids were seen in eight patients (25%). CONCLUSIONS: In our experience, 25% of the desmoid tumors (8/32) were recurrent and postoperative radiotherapy did not seem to influence the local recurrence rate. The most important predictor for recurrence was tumors of > 5 cm. PMID- 19282565 TI - Prognostic significance of bone marrow histology in multiple myeloma. AB - BACKGROUND: Bone marrow examination continues to be the cornerstone for establishing the diagnosis of multiple myeloma in association with other clinical and laboratory parameters. Plasma cell morphology has significant correlation with clinical stage and survival. AIMS: To note the bone marrow histology in detail in multiple myeloma and to correlate it with clinical stage and survival. METHODS AND MATERIAL: Fifty-five cases of multiple myeloma diagnosed between January 2001 and December 2006, who had a bone marrow aspiration and biopsy done at the time of diagnosis were included in the present study. STATISTICAL ANALYSIS: SPSS software version 13.0 was used. Clinical stage and plasma cell morphology were correlated using chi square test and Spearman's correlation coefficient. Survival analysis was done using the Kaplan-Meier method. RESULTS: Seventy-six percent patients were in clinical stage III, 17% and 7% were in stage II and I respectively. The clinical stage correlated significantly with plasma cell morphology, percentage of plasma cell infiltration and pattern of infiltration. Plasma cell morphology correlated significantly with bone marrow parameters like percentage infiltrate, pattern of infiltration, degree of fibrosis and mitotic activity. Patients in advanced clinical stage,> 50% plasma cells in the marrow, diffuse pattern of infiltration, high mitosis and increased fibrosis had a shorter median survival than patients with favorable features. CONCLUSIONS: It is recommended that the bone marrow histology be studied in detail in multiple myeloma at diagnosis since it correlates well with the clinical stage and offers useful prognostic information. PMID- 19282566 TI - Primary testicular non-Hodgkin lymphoma: a single institution experience from India. AB - BACKGROUND: Primary testicular non-Hodgkin lymphoma (NHL) is an uncommon extra nodal presentation, constituting 1% of all NHL. Median age at time of presentation is 60 years. Anthracycline based chemotherapies are most frequently used. There is not enough data on use of monoclonal antibody (Rituximab) in testicular NHL. METHODS: We screened approximately eight hundred and fifty NHL cases registered from January 2002 to May 2008 and found six primary testicular NHL patients. These six cases were analyzed for baseline clinical features, investigations, staging, treatment and outcome variables. RESULTS: Median age was 55 years (range 7-72 years) and median duration of symptoms was 3.5 months (range 1-8 months). All patients had testicular swelling and abdominal lymphadenopathy. Five patients (83%) had stage IV and one had stage IE disease. Majority had diffuse large B-cell histology (83%). All patients were treated with anthracycline based combination chemotherapy and CNS prophylaxis after local therapy except one pediatric patient who did not receive any local therapy. Four patients completed therapy and are on follow up while two patients having extensive disease with poor performance status died of neutropenic sepsis after 1 2 cycles of chemotherapy. The surviving four patients achieved complete remission and are without any recurrence with a median follow up of 26 months (1-78 months). CONCLUSION: Primary testicular NHL is an uncommon entity and with current combined modality treatment and CNS prophylaxis, the outcome may be as good as nodal NHL. PMID- 19282567 TI - A comparative study of low dose weekly paclitaxel versus cisplatin with concurrent radiation in the treatment of locally advanced head and neck cancers. AB - PURPOSE: The purpose of this study was to compare low dose weekly paclitaxel versus cisplatin with concurrent radiation in locally advanced head and neck cancers. MATERIALS AND METHODS: From August 2005 to July 2006, a total of 100 biopsy proven, locally advanced head and neck cancers were enrolled for the study. All the patients were stratified in two groups, study group A and control group B. Study group patients received injection Paclitaxel 20 mg/m 2 , I/V 1 hr infusion weekly for 6 weeks and control group patients received injection Cisplatin 30 mg/m 2 , I/V 2 hrs infusion weekly for 6 weeks. All patients received 66-70 Gy concurrent radiation at the rate of 2 Gy/day, 5 #/week, in 6-7 weeks by cobalt theratron phoenix - 80 teletherapy units. RESULT: Complete response achieved in 73% of patients in study group and 64% of patients in control group. There was no statistically significant difference observed between the study group and the control group (chi2 = 1.167, df = 1, level of significance 0.05). On 3-10 months of follow-up 59% of patients in the study group and 42% of patients in the control group are alive and disease free. Local toxicities including mucositis, dysphasia and skin reactions were more in the study group but tolerable. CONCLUSION: Efficacy of paclitaxel in low dose weekly schedule is comparable to cisplatin in locally advanced head and neck squamous cell carcinoma. Further analysis and follow-up are needed to evaluate if this benefit will translate into prolonged survival. PMID- 19282568 TI - Survival effects of cyclooxygenase-2 and 12-lipooxygenase in Egyptian women with operable breast cancer. AB - BACKGROUND: Breast cancer (BC) is the commonest among women in Egypt as well as in many other countries. Cyclo-oxygenase-2 (COX-2) and 12-lipo-oxygenase (12-LOX) are over-expressed in 30-40% of patients and carry a poor prognosis. The objectives of this study were to correlate COX-2 and 12-LOX expression with various clinico-pathologic patients' characteristics and their impact on overall survival (OS) and disease free survival (DFS) in Egyptian women with operable BC. MATERIALS AND METHODS: This prospective study included 57 consecutive BC cases presenting to the Egyptian National Cancer Institute. Sections from BC and nearby normal tissues were examined for expression of COX-2 and 12-LOX using reverse transcriptase polymerase chain reaction. RESULTS: The patients' median age was 45 years. Fifty-three percent were premenopausal. Stage II and III disease represented 25 and 75% respectively. Adjuvant chemotherapy, radiotherapy and tamoxifen were used in 90, 75 and 60% respectively. Sixty percent had hormone receptor positive tumors and 28% over-expressed HER2/neu. Forty-nine and sixty five percent showed over-expression of COX-2 and 12-LOX respectively. Patients with higher TNM stage or who developed visceral metastases had significantly higher COX-2 expression. For the whole group of patients, the median DFS was 37 months, while the median OS was not reached. OS or DFS did not differ significantly between patients with normal and over-expression of COX-2. DFS but not OS was significantly higher in 12-LOX over-expression compared to normal expression. CONCLUSION: COX-2 over-expression was associated with poor prognostic criteria in BC, but did not affect DFS or OS. 12-LOX over-expression was associated with better DFS, but not OS. PMID- 19282569 TI - Coexistent hepatic and pulmonary epitheloid hemangioendothelioma. AB - Epitheloid hemangioendothelioma (EHE) is a rare neoplasm of vascular origin known to arise in soft tissue, liver and lung. We describe a case of coexistent hepatic and pulmonary epitheloid hemangioendothelioma, proven on autopsy, and review the histological and radiological features of epitheloid hemangioendothelioma. The coexistence of hepatic with pulmonary EHE has been reported in only a few cases. Large confluent masses, peripheral location with capsular retraction, hypertrophy of uninvolved liver, invasion of portal and hepatic veins, enhancing margins and delayed enhancement and dense calcification are the typical features which provide a clue to diagnosis of hepatic EHE. In patients with both hepatic and pulmonary EHE it is difficult to say whether the tumor arose primarily in the lung or liver, or began simultaneously in both organs. PMID- 19282570 TI - Recurrent alpha-fetoprotein secreting Sertoli-Leydig cell tumor of ovary with an unusual presentation. AB - Alpha-fetoprotein secreting (AFP) Sertoli-Leydig cell tumors of ovary (SLCT) are now identified as a distinct entity among the uncommon group of sex cord tumors of ovary. We report an unusual case of recurrent AFP secreting ovarian tumors and as ileocecal mesenteric cyst in a 25-year-old patient resulting in difficulty in initial diagnosis of AFP producing SLCT. Although six recurrent cases were described out of the 25 reported cases of AFP secreting SLCTs, this patient with an unusual presentation of recurrence is the second case in the literature to the best of our knowledge. PMID- 19282571 TI - Mucosal malignant melanoma of the nasal cavity. AB - Mucosal malignant melanoma (MMM) of the nose is extremely rare. We report a case of MMM of the nasal cavity in a 60-year-old male patient presenting with a polypoidal mass in the right nasal cavity. It was increasing gradually and obstructing breathing. A biopsy of the lesion was done with a clinical suspicion of inverted papilloma/carcinoma. Microscopy revealed features suggestive of malignant melanoma with minimal melanin pigmentation. Subsequently wide local excision was done. Diagnosis of malignant melanoma was facilitated by histochemistry and immunohistochemistry. PMID- 19282572 TI - Transitional cell carcinoma urinary bladder manifesting as second malignancy following radiotherapy for carcinoma cervix. PMID- 19282573 TI - Rare occurrence of carcinoma esophagus in a case of epidermolysis bullosa. PMID- 19282574 TI - Changing trends in incidence of breast cancer: Indian scenario. PMID- 19282575 TI - Castleman's disease: a rare diagnosis in retroperitoneum. PMID- 19282576 TI - Refractory hypokalemia due to conventional amphotericin B in patients with leukemia. PMID- 19282577 TI - Colon cancer awareness for prevention: Call for global initiatives. PMID- 19282578 TI - Acne in India: guidelines for management - IAA consensus document. PMID- 19282604 TI - Molecular basis of restenosis and novel issues of drug-eluting stents. AB - Restenosis after stent deployment is an overreaction of the wound healing response after vascular injury, and is characterized by the sequence of inflammation, granulation, extracellular matrix remodeling, and smooth muscle cell (SMC) proliferation and migration. In contrast, reendothelialization of at least part of the injured vessel surface, which is essential in the wound healing process, may occur at the site of stenting. Recent advances in drug-eluting stents (DES) have substantially reduced restenosis, but do not contribute to improve long-term prognosis, compared with bare metal stents (BMS). One of the reasons may be that reendothelialization is impaired after DES stenting. Regenerated endothelial cells and proliferated SMCs in the neointima are both in part derived from their progenitor cells, which are mobilized from bone marrow to injured vessel sites and differentiate into both vascular endothelial cells and SMCs. DES inhibits mobilization and differentiation of endothelial and smooth muscle progenitor cells, and thus not only inhibits restenosis but also impairs reendothelialization, which may lead to late stent thrombosis. To improve long term prognosis in the DES era, adjunctive medical treatments inducing early reendothelialization, but inhibiting SMC proliferation, would be required. PMID- 19282605 TI - Silent myocardial ischemia. AB - Although much progress has been made in reducing mortality from ischemic cardiovascular disease, this condition remains the leading cause of death throughout the world. This might in part be due to the fact that over half of patients have a catastrophic event (heart attack or sudden death) as their initial manifestation of coronary disease. Contributing to this statistic is the observation that the majority of myocardial ischemic episodes are silent, indicating an inability or failure to sense ischemic damage or stress on the heart. This review examines the clinical characteristics of silent myocardial ischemia, and explores mechanisms involved in the generation of angina pectoris. Possible mechanisms for the more common manifestation of injurious reductions in coronary flow; namely, silent ischemia, are also explored. A new theory for the mechanism of silent ischemia is proposed. Finally, the prognostic importance of silent ischemia and potential future directions for research are discussed. PMID- 19282606 TI - Predictive value of C-reactive protein for major postoperative complications following off-pump coronary artery bypass surgery: prospective and observational trial. AB - BACKGROUND: To prospectively investigate the predictive value of the preoperative C-reactive protein (CRP) concentration for major postoperative complications following off-pump coronary artery bypass (OPCAB) surgery. METHODS AND RESULTS: From January 2007 to December 2007, 185 consecutive patients scheduled for elective OPCAB surgery were allocated to a low-CRP group (n=137, CRP <0.3 mg/dl) and a high-CRP group (n=48, CRP > or = 0.3 mg/dl). The incidence of major postoperative complications, defined as postoperative myocardial infarction, and 5 major morbidity endpoints including permanent stroke, renal dysfunction, any cardiac surgery reoperation, ventilation for more than 48 h, and deep sternal wound infection were assessed and compared. Multivariate logistic regression was used to determine the predictors of major postoperative complications. Patients in the high-CRP group had a significantly higher overall incidence of major postoperative complications, particularly renal dysfunction. In the multivariate logistic regression model, adjusting all the significant univariate predictors, baseline CRP >0.3 mg/dl and preoperative chronic renal failure (CRF) remained as significant independent predictors of major postoperative complications. CONCLUSIONS: Elevated preoperative CRP level and/or preoperative CRF indicate increased risk of developing major postoperative complications, particularly acute postoperative renal dysfunction in patients undergoing OPCAB surgery. PMID- 19282607 TI - Ten-year patency and factors causing restenosis after endovascular treatment of iliac artery lesions. AB - BACKGROUND: Long-term outcome and the factors associated with restenosis after endovascular treatment (EVT) for iliac artery lesions in peripheral arterial disease (PAD) were evaluated. METHODS AND RESULTS: EVT was performed for 487 lesions (TransAtlantic Inter-Society Consensus-II (TASC-II) Type-A: 275, B: 115, C: 37, and D: 60) in 436 PAD patients. The initial success rates for Type-B and Type-D lesions were lower than for Type-A lesions (P<0.05). The 3-, 5- and 10 year patency rates were 67%, 54% and 50%, respectively, with plain-old balloon angioplasty (POBA), and 88%, 82% and 75%, respectively, for stenting after suboptimal POBA, showing a significantly higher patency after treatment with a stent (P<0.001). With POBA, the long-term patency for Type-C/D lesions was lower than for Type-A/B lesions (P<0.05), but the patency after stenting did not differ significantly between Type-C/D and A/B. In the univariate analysis, the TASC-II classification, lesion length, pre- and post-procedural stenosis rates and stent use were found to be significant factors associated with restenosis (P<0.05). In the multivariate analysis, stent use (hazard ratio (HR) 0.345, confidence interval (CI) 0.193-0.616, P<0.001) and the post-procedural stenosis rate (HR 1.015, CI 1.001-1.030, P<0.05) were significantly associated with restenosis. CONCLUSIONS: Stent use and a low residual stenosis rate are significantly associated with patency, and favorable long-term patency can be obtained with stent placement for selected TASC-II Type-C/D lesions. PMID- 19282608 TI - New access for radiofrequency catheter ablation of left-sided atrioventricular accessory pathways: safety and efficacy of the transradial approach. AB - BACKGROUND: The safety and efficacy of the transradial approach for radiofrequency catheter ablation (RFCA) of left-sided atrioventricular accessory pathways (APs) was evaluated in the present study. METHODS AND RESULTS: Included were 40 consecutive patients with type A Wolff-Parkinson-White (WPW) syndrome who underwent RFCA via the radial artery route, and 30 patients with type A WPW syndrome who underwent RFCA via the transfemoral approach (controls) were retrospectively chosen for control. All 45 APs in the 40 patients were successfully ablated: 35 APs were successfully blocked with 1 ablation attempt, and the other 10 APs were ablated after 2-4 attempts. Compared with the transfemoral approach, the total procedure time for the transradial approach was longer (40 +/- 7.7 vs 32.4 +/- 8.7 min, P<0.05) and the fluoroscopic time was similar (7.2 +/- 2.2 vs 7.9 +/- 3.9 min, P>0.05). There were no vascular complications in the transradial group, but 2 patients in the transfemoral developed local hematoma. There was no recurrence of arrhythmia in either group. CONCLUSIONS: The transradial approach is a safe and effective access for RFCA of left-sided APs. PMID- 19282609 TI - Metabolic syndrome and all-cause and cardiovascular disease mortality: Japan Public Health Center-based Prospective (JPHC) Study. AB - BACKGROUND: Although the metabolic syndrome (MetS) is considered to be caused primarily by visceral fat accumulation, epidemiological evidence is lacking as to whether or not obesity is an essential element in the syndrome. METHODS AND RESULTS: Between 1990 and 2005, the Japan Public Health Center-based Prospective (JPHC) Study conducted baseline measurements of metabolic risk factors in 12,412 men and 21,639 women, aged 40-69 years, with no history of cardiovascular disease (CVD) or cancer. To clarify the role of obesity, which the definition of MetS in Japan has adopted as an essential criterion, clustering of risk factors in data grouped according to overweight condition was examined. During a 12.3-year follow up there were 2,040 deaths, including 947 from cancers and 304 from CVD. MetS significantly increased the hazard ratios for all-cause mortality in women and CVD mortality in men. Non-overweight with > or = 2 risk factors had a similar impact on all-cause and CVD mortality. Clustering of metabolic factors caused a linear increase in the hazard ratios for mortality. CONCLUSIONS: MetS caused moderate increases in all-cause and CVD mortality. However, the MetS definition requiring obesity may not necessarily identify non-overweight individuals who have a high mortality risk and are more prevalent than subjects with MetS. PMID- 19282610 TI - Effect of cilostazol treatment on adiponectin and soluble CD40 ligand levels in diabetic patients with peripheral arterial occlusion disease. AB - BACKGROUND: Peripheral arterial occlusion disease (PAOD) is caused mainly by chronic inflammation and endothelium dysfunction, and is often treated with cilostazol. However, because this drug's influence on atherogenic cytokines is still not well known, this study examined the effect of cilostazol on the serum levels of soluble CD40 ligand (sCD40L), adiponectin and high-sensitivity C reactive protein (hs-CRP) in patients with type 2 diabetes and PAOD. METHODS AND RESULTS: The 92 type 2 diabetics with PAOD and 100 non-PAOD diabetics were enrolled and randomly assigned to a group receiving either cilostazol or placebo for 6 months. The atherogenic cytokines were measured at the beginning and completion of the study. In the PAOD groups, those in the cilostazol group had significant changes in the levels of hs-CRP, sCD40L and adiponectin (P=0.001, P=0.05, P=0.004, respectively). Changes in the levels of adiponectin and sCD40L were more significant in the PAOD group treated with cilostazol than in the non PAOD group also treated with the drug (P=0.01 and P=0.008, respectively). CONCLUSIONS: Cilostazol can decrease hs-CPR and sCD40L levels and increase that of adiponectin, and then delay the progression of atherogenesis and chronic inflammation in type 2 diabetics, especially those with PAOD. PMID- 19282611 TI - Effects of cardiac complications on cardiovascular events in Japanese high-risk hypertensive patients: subanalysis of the CASE-J trial. AB - BACKGROUND: The Candesartan Antihypertensive Survival Evaluation in Japan (CASE J) trial compared the effects of candesartan and amlodipine on cardiovascular events in Japanese high-risk hypertensive patients. The present study aimed to clarify the effect of cardiac complications on cardiovascular events in patients enrolled in CASE-J. METHODS AND RESULTS: Cardiac complications were defined as left ventricular hypertrophy (LVH) and ischemic heart disease (IHD). The primary endpoint was a composite of sudden death, cerebrovascular, cardiac, renal and vascular events. The study group was divided into 2,030 and 2,673 patients with and without cardiac complications. During 3.2 follow-up years, cardiovascular events occurred for a rate of 13.6 per 1000 patient-years in patients without cardiac complications, and 23.1 per 1000 patient-years in patients with cardiac complications (adjusted hazard ratio (HR): 2.22; P<0.001). Furthermore, LVH was associated with the onset of cerebrovascular events (adjusted HR: 2.38; P<0.001), whereas IHD was associated with the onset of cardiovascular death (adjusted HR: 2.22; P<0.05), especially sudden death and other cardiac events. CONCLUSIONS: Cardiac complications are independent predictors for cardiovascular events in Japanese high-risk hypertensive patients. In particular, LVH is related to cerebrovascular events and IHD is related to cardiac death and other cardiac events. PMID- 19282612 TI - Infiltration of macrophages through the atrial endocardium of inflammation induced rats: contribution of fractalkine. AB - BACKGROUND: Inflammatory processes in the atria during systemic inflammation remain unclear, so this study tested the hypothesis that macrophages infiltrate the atrial myocardium mainly through the atrial endocardium with the contribution of fractalkine. METHODS AND RESULTS: Sprague-Dawley rats were injected with lipopolysaccharide (LPS) to simulate inflammation in the atria. Inflammation was immunohistologically assessed by the presence of macrophages. Macrophage infiltration was diffuse throughout the atrial myocardium after LPS injection. At an earlier phase after LPS injection, the number of macrophages dramatically increased, mainly in the atrial endocardium, and the expression of fractalkine protein was markedly increased by treatment with LPS in the atrial endocardium. The LPS-induced increase in atrial macrophage infiltration was significantly suppressed by neutralizing the fractalkine protein (P<0.01). CONCLUSIONS: In an experimental model of atrial inflammation, macrophages infiltrated the myocardium mainly through the atrial endocardium with the contribution of fractalkine. Inhibition of macrophage infiltration by suppressing chemokine expression could be a novel therapeutic approach to controling acute inflammation in the atria. PMID- 19282613 TI - [Molecular mechanisms of abnormal vascular contraction and the screening for their molecular-targeted therapeutic drugs]. PMID- 19282614 TI - [Direct regulation of contractile filaments: novel therapeutic strategy for vasospasm treatment]. PMID- 19282615 TI - [Roles of Rho/Rho-kinase signaling in hypertension and pulmonary hypertension]. PMID- 19282616 TI - [Therapeutic neovascularization by nanotechnology-mediated drug delivery system]. PMID- 19282617 TI - [The non-kinase activity of myosin light chain kinase in regulating smooth muscle contraction]. PMID- 19282618 TI - [Pharmaceutical research in the early stages of drug discovery--significance and practice]. PMID- 19282619 TI - [Evaluation methods for nephrotoxicity]. PMID- 19282620 TI - [Levobupivacaine hydrochloride injection (POPSCAINE): pharmacological characteristics and clinical study result]. PMID- 19282621 TI - Involvement of PlsX and the acyl-phosphate dependent sn-glycerol-3-phosphate acyltransferase PlsY in the initial stage of glycerolipid synthesis in Bacillus subtilis. AB - The gene responsible for the first acylation of sn-glycerol-3-phosphate (G3P) in Bacillus subtilis has not yet been determined with certainty. The product of this first acylation, lysophosphatidic acid (LPA), is subsequently acylated again to form phosphatidic acid (PA), the primary precursor to membrane glycerolipids. A novel G3P acyltransferase (GPAT), the gene product of plsY, which uses acyl phosphate formed by the plsX gene product, has recently been found to synthesize LPA in Streptococcus pneumoniae. We found that in B. subtilis growth arrests after repression of either a plsY homologue or a plsX homologue were overcome by expression of E. coli plsB, which encodes an acyl-acylcarrier protein (acyl-ACP) dependent GPAT, although in the case of plsX repression a high level of plsB expression was required. B. subtilis has, therefore, a capability to use the acyl ACP dependent GPAT of PlsB. Simultaneous expression of plsY and plsX suppressed the glycerol requirement of a strict glycerol auxotrophic derivative of the E. coli plsB26 mutant, although either one alone did not. Membrane fractions from B. subtilis cells catalyzed palmitoylphosphate-dependent acylation of [14C]-labeled G3P to synthesize [14C]-labeled LPA, whereas those from DeltaplsY cells did not. The results indicate unequivocally that PlsY is an acyl-phosphate dependent GPAT. Expression of plsX corrected the glycerol auxotrophy of a DeltaygiH (the deleted allele of an E. coli homologue of plsY) derivative of BB26-36 (plsB26 plsX50), suggesting an essential role of plsX other than substrate supply for acyl phosphate dependent LPA synthesis. Two-hybrid examinations suggested that PlsY is associated with PlsX and that each may exist in multimeric form. PMID- 19282622 TI - A novel epistatic interaction at two loci causing hybrid male sterility in an inter-subspecific cross of rice (Oryza sativa L.). AB - Postzygotic reproductive isolation (RI) often arises in inter-subspecific crosses as well as inter-specific crosses of rice (Oryza sativa L.). To further understand the genetic architecture of the postzygotic RI, we analyzed genes causing hybrid sterility and hybrid breakdown in a rice inter-subspecific cross. Here we report hybrid male sterility caused by epistatic interaction between two novel genes, S24 and S35, which were identified on rice chromosomes 5 and 1, respectively. Genetic analysis using near-isogenic lines (NILs) carrying IR24 (ssp. indica) segments with Asominori (ssp. japonica) genetic background revealed a complicated aspect of the epistasis. Allelic interaction at the S24 locus in the heterozygous plants caused abortion of male gametes carrying the Asominori allele (S24-as) independent of the S35 genotype. On the other hand, male gametes carrying the Asominori allele at the S35 locus (S35-as) showed abortion only when the IR24 allele at the S24 locus (S24-ir) was concurrently introgressed into the S35 heterozygous plants, indicating that the sterility phenotype due to S35 was dependent on the S24 genotype through negative epistasis between S24-ir and S35 as alleles. Due to the interaction between S24 and S35, self-pollination of the double heterozygous plants produced pollen-sterile progeny carrying the S24 ir/S24-ir S35-as/S35-ir genotype in addition to the S24 heterozygous plants. This result suggests that the S35 gene might function as a modifier of S24. This study presents strong evidence for the importance of epistatic interaction as a part of the genetic architecture of hybrid sterility in rice. In addition, it suggests that diverse systems have been developed as postzygotic RI mechanisms within the rice. PMID- 19282623 TI - High-throughput evaluation of aryl hydrocarbon receptor-binding sites selected via chromatin immunoprecipitation-based screening in Hepa-1c1c7 cells stimulated with 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - Upon binding to ligands such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), an aryl hydrocarbon receptor (AhR) is activated to form a heterodimer with an aryl hydrocarbon receptor nuclear translocator (Arnt) and binds to DNA. It has been shown that the binding of AhR to DNA depends on the dioxin response element (DRE) and controls xenobiotic-response genes. AhR-binding DNA fragments from mouse hepatoma Hepa-1c1c7 cells stimulated with TCDD were once enriched in a chromatin immunoprecipitation (ChIP) DNA library and screened through a high-throughput southwestern chemistry-based enzyme-linked immunosorbent assay (SW-ELISA). After screening 1700 fragments, the ChIP-SW-ELISA screening strategy allowed us to isolate 77 fragments tightly interacting with AhR in the presence of TCDD. Only 39 of the 77 fragments appeared to contain a typical DRE, indicating that in some cases the DRE was dispensable for AhR-binding, while 75 fragments were located within promoter-distal regions. Genomic mapping of the 77 fragments enabled us to estimate 121 potential AhR targets including known targets such as Cyp1A1 and Cyp1B1, but only a limited number exhibited an altered expression dependent on TCDD. This study revealed the fact that TCDD-activated AhR frequently binds to promoter-distal regions even without a DRE and is not always involved in transcriptional regulation, suggesting that within the genome DNA-binding of AhR could take place often in many regions without cis-regulatory elements and might not be a key determinant to establish its regulatory function. PMID- 19282624 TI - Spatial and temporal control of transcription of the soybean beta-conglycinin alpha subunit gene is conferred by its proximal promoter region and accounts for the unequal distribution of the protein during embryogenesis. AB - Differentiation into specific embryo cell types correlates with the processes that lead to the accumulation of seed storage proteins in plants. The alpha subunit of beta-conglycinin, a major component of seed storage proteins in soybean, accumulates at a higher level in cotyledons than in the embryonic axis in developing embryos. To understand the mechanisms underlying this phenomenon, we characterized the upstream region of the alpha subunit gene in terms of transcriptional control using transgenic Arabidopsis thaliana plants carrying reporter gene constructs comprising the 1357-bp upstream sequence of the alpha subunit gene and the beta-glucuronidase (GUS) gene. Analysis of the time-course dependent pattern of GUS expression revealed that the expression was first confined to the cotyledons and occurred later in the entire embryo during embryogenesis. The level of GUS expression was higher in cotyledons than in the embryonic axis throughout the period of its expression, coincident with the distribution of the alpha subunit protein in soybean embryos. By testing progressively shorter promoter fragments, the cis-acting elements responsible for transcriptional activation in the cotyledons and the embryonic axis were both localized to the region spanning -245 to -161 relative to the transcription start site. It is also concluded that the upstream region up to -245 is sufficient to control the spatial and temporal pattern of transcription, while further upstream regions influence transcription rate without affecting the transcriptional pattern. Overall, these results indicate that the unequal distribution of alpha subunit protein within the embryos is established primarily as a consequence of differential transcriptional activation controlled by a short proximal promoter region of the gene in different embryonic tissues. PMID- 19282627 TI - Azospirillum palatum sp. nov., isolated from forest soil in Zhejiang province, China. AB - A Gram-negative bacterium designated ww 10(T) was isolated by plating dilutions from forest soil in Zhejiang province, China. Strain ww 10(T) was investigated by polyphasic taxonomic study including phenotypic and phylogenetic analysis. Cells of ww 10(T) were Gram-negative, strictly aerobic, motile with peritrichous flagella and rod-shaped. The strain grew optimally at 30-37 masculineC and pH 6.0 8.0. The major fatty acids were C(18:1)omega7C, cyclo-C(19:0) omega8C and C(16:0). The respiratory quinones contained a large amount of Q-10, a moderate component of Q-9 and a minor of Q-8. The G+C content of genomic DNA was about 67.3 mol%. Phylogenetic analysis revealed that strain ww 10(T) belongs to the phyletic cluster of genus Azospirillum and displayed 16S rRNA gene sequence similarity lower than 96.3% to the four closest described species of the genera Azospirillum and Roseomonas. Results of polyphasic taxonomic analysis showed that strain ww 10(T) represents a novel species in the genus Azospirillum, for which the name Azospirillum palatum sp. nov., is proposed. The type strain is ww 10(T) (=LMG 24444(T)=KCTC 13200(T)=CCTCC AB 207189(T)). PMID- 19282628 TI - Development of AFLP markers and phylogenetic analysis in Hypsizygus marmoreus. AB - The present study was designed to develop a reliable experimental protocol for detecting and genotyping amplified fragment length polymorphism (AFLP) markers in the commercially important edible mushroom Hypsizygus marmoreus; and to use the markers to evaluate the genetic polymorphisms and phylogenetic relationships among 19 strains of H. marmoreus. Using a set of ten primer pairs, 609 AFLP markers were identified, of which 532 were found segregating among 19 cultivated varieties and laboratory stock strains. An estimate of genetic relationships among the varieties/strains based on the AFLP markers was obtained by primary phylogenetic analysis using UPGMA. Our data demonstrate abundant genetic variation in the natural population of H. marmoreus and facilitate further analysis of genetic divergence within the species. The present study is useful for understanding the genetic structure of this species and uncovering genetic control of economically important traits for efficient breeding programs. PMID- 19282629 TI - Kitasatospora saccharophila sp. nov. and Kitasatospora kazusanensis sp. nov., isolated from soil and transfer of Streptomyces atroaurantiacus to the genus Kitasatospora as Kitasatospora atroaurantiaca comb. nov. AB - A polyphasic study was undertaken to establish the taxonomic positions of two isolates, SK15(T) and SK60(T), from soil samples that were found to have morphological and chemical properties consistent with Kitasatospora strains. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strains SK15(T) and SK60(T) form novel evolutionary lineages within the radiation of the genus Kitasatospora and share the highest 16S rRNA gene sequences similarities with their closest relatives, Kitasatospora setae IAM 15325(T) (97.8%) and Kitasatospora mediocidica IAM 15162(T )(97.5%), respectively. However, the results of DNA-DNA hybridization experiment and phenotypic data demonstrated that strains SK15(T) and SK60(T) are distinct from their closest phylogenetic neighbors and other Kitasatospora species. For chemotaxonomic characteristics, the cell-wall peptidoglycan of strains contained both meso- and LL-diaminopimelic acids as the diamino acids, the predominant quinone system was MK-9(H(6)) and MK 9(H(8)), whole-cell hydrolysates were rich in galactose, mannose and ribose, and the major fatty acids were C(16:0), anteiso-C(15:0), iso-C(15:0) and iso-C(16:0). On the basis of both phenotypic and phylogenetic evidence, strains SK15(T) and SK60(T) were assigned to represent two novel species of the genus Kitasatospora, for which the names Kitasatospora saccharophila sp. nov. (type strain SK15(T)=JCM 14559(T)=KCTC 19566(T)) and Kitasatospora kazusanensis sp. nov. (type strain SK60(T)=JCM 14560(T)=KCTC 19565(T)) are proposed. It is also proposed that Streptomyces atroaurantiacus should be transferred to the genus Kitasatospora as Kitasatospora atroaurantiaca comb. nov. (type strain NBRC 14327(T)=DSM 41649(T)). PMID- 19282630 TI - Biosorptive capacity of Cd(II) and Cu(II) by lyophilized cells of Pseudomonas stutzeri. AB - The biosorptive capacity of Cd(II) and Cu(II) by lyophilized cells of Pseudomonas stutzeri was investigated based on Langmuir and Freundlich isotherms and biosorption kinetics were analyzed using first order kinetic with different initial metal concentrations. Biosorptive capacity for Cd(II) and Cu(II) decreased with an increment of metal concentration, reaching 43.5 and 36.2 mg/g at the initial concentration of 300 mg/L. Biosorption capacity for both metal ions was increased with increasing pH. The optimum pH for biosorption rate of Cu(II) and Cd(II) was pH 5; above pH 5.0 the metal cations came to be precipitated. The experimental data showed a better fit with the Langmuir model over the Freundlich model for both metal ions throughout the range of initial concentrations. The maximum sorptive capacity (q max) obtained from the Langmuir equation for Cd(II) and Cu(II) were 47.86 (r(2)=0.99) and 33.16 (r(2)=0.99), respectively. The bacterial cells have more affinity to adsorb cadmium than copper. The first order kinetic was well fitted to the experimental data for initial concentrations from 30 to 100 mg/L during reaction times of 250 min. These results suggest that biosorption of Cu(II) and Cd(II) by lyophilized cells of P. stutzeri is a potential metal removal strategy. PMID- 19282631 TI - Ability of Agrobacterium tumefaciens and A. rhizogenes strains, inability of A. vitis and A. rubi strains to adapt to salt-insufficient environment, and taxonomic significance of a simple salt requirement test in the pathogenic Agrobacterium species. AB - Resistance to a 1% or higher concentration of NaCl is an important trait for taxonomic discrimination of species in the family Rhizobiaceae. However, we have little knowledge about how much salt rhizobia require. In this study, we examined the requirement of NaCl for growth in relation to the NaCl sensitivity in the pathogenic Agrobacterium species. Consistent with the previous salt resistance data, the standard Luria Bertani medium containing 0.5% NaCl (LB) permitted A. tumefaciens and A. vitis strains to grow well, but not A. rhizogenes strains. In contrast, LB lacking NaCl (LB-NaCl) allowed the A. rhizogenes and A. tumefaciens strains to grow well but not the A. vitis strains. In LB-NaCl, viability of A. vitis strains decreased 500-fold in 24 h. The addition of KCl, MgCl(2) or MgSO(4) to LB-NaCl restored the growth of A. vitis strains. These data indicate higher salt requirements in A. vitis than those in A. tumefaciens and A. rhizogenes and adaptability of A. tumefaciens to salt-insufficient environments. An A. rubi strain was salt dependent like A. vitis. The experiment was extended to strains in related genera. Checking growth on the two media was very easy, gave a new trait and clear results, and thereby proved useful as an additional method for taxonomic identification. PMID- 19282632 TI - Rhodovastum atsumiense gen. nov., sp. nov., a phototrophic alphaproteobacterium isolated from paddy soil. AB - A photoorganotrophic alphaproteobacterium designated strain G2-11(T) was isolated from submerged paddy soil. This bacterium had relatively large, oval to rod shaped cells (2.0-3.0x3.0-10 microm). Cells were motile by means of single polar flagella. The color of phototrophically growing cultures was reddish-brown. The cell extract had absorption maxima at 375, 465, 492, 529, 592, 804, and 844 nm, indicating the presence of bacteriochlorophyll a and carotenoides of the spirilloxanthin series. Vesicular intracytoplasmic membranes were present. The main component of cellular fatty acids was C(18:1)omega7c. Ubiquinone-10 and rhodoquinone-10 were the major quinones. A 16S rRNA gene sequence analysis revealed that the isolate is closest to the acidophilic aerobic photosynthetic bacterium Acidisphaera rubrifaciens strain HS-AP3(T) (93.3% similarity). The G+C content of genomic DNA is 67.8 mol%. The name Rhodovastum atsumiense gen. nov., sp. nov. is proposed for the novel isolate. The type strain is strain G2-11(T) (=NBRC 104268(T)=KCTC 5708(T)). PMID- 19282633 TI - Isolation and characterization of Shewanella oneidensis WL-7 capable of decolorizing azo dye Reactive Black 5. PMID- 19282634 TI - Immunoprotective analysis of two Edwardsiella tarda antigens. PMID- 19282635 TI - Chemical composition and biological activities of Jeju Thymus quinquecostatus essential oils against Propionibacterium species inducing acne. PMID- 19282636 TI - Diversity of heterotrophic bacteria isolated from biofilm samples and cell surface hydrophobicity. PMID- 19282637 TI - Nitrate removal mediated by soil microorganism, Enterobacter sp. GG0461. PMID- 19282638 TI - Aroma-active compounds of Elatostema laetevirens and Elatostema umbellatum var. majus. AB - The essential oils from aerial parts of Elatostema laetevirens and Elatostema umbellatum var. majus were investigated by capillary GC and GC-MS. The important aroma-active compounds were also detected in the oil using GC-MS/O and aroma extraction dilution analysis (AEDA). As a result, 79 compounds of E. laetevirens and 80 compounds of E. umbellatum var. majus, accounting for 95.41% and 98.37%, were identified, respectively. The major components of E. laetevirens oil were phytol (667.4 microg/100 g fresh samples), neophytadiene (335.2 microg) and gamma himachalene (49.8 microg). On the other hand, E. umbellatum var. majus oil contained phytol (402.3 microg), linoleic acid (289.4 microg) and palmitic acid (241.4 microg) as the major components. GC-MS/O and AEDA showed that (2E)-hexenal and (2E, 4E)-nonadienal were most aroma compounds of E. laetevirens oil. It seems that these components make the green-floral odor. On the other hands, it seems that (2E)-hexenal, (3Z)-hexenol and 1-octen-3-ol make the green-oily odor of E. umbellatum var. majus oil. PMID- 19282639 TI - Linoleic acid-menthyl ester reduces the secretion of apolipoprotein B100 in HepG2 cells. AB - The effect of linoleic acid-menthyl ester (LAME) on lipid metabolism were assessed in HepG2 cells. It is well known that high level of apolipoprotein (apo) B100 in the serum is risk for atherosclerosis. Although linoleic acid (LA) treatment and LA plus L-mentol treatment increased apo B100 secretion, LAME treatment significantly decreased apo B100 secretion in HepG2 cells compared with control medium. The hypolipidemic effect of LAME was attributable to the suppression of triglyceride synthesis in HepG2 cells. It is also known that the risk of coronary heart disease is negatively related to the concentration of serum apo A-1. In the present study, LAME treatment increased apo A-1 secretion as compared with LA treatment in HepG2 cells. These results suggest that mentyl esterification of fatty acids may be beneficial in anti-atherogenic dietary therapy. PMID- 19282640 TI - Partition coefficients of 1-alkanols between water and DDAB vesicle membrane determined by differential conductivity method. AB - The partition coefficients of 1-alkanols between water and didodecyldimethylammonium bromide (DDAB) vesicle membrane, and the change of the degree of counter-ion dissociation on DDAB vesicle caused by the solubilization of 1-alkanols were determined from electric conductivity measurements. A good linear relationship was obtained between logarithmic value of the partition coefficient and carbon number of alkyl chain in 1-alkanols. The standard transfer free energy change per methylene group was evaluated from the slope of this straight line. On the other hand, the effect of 1-alkanols on the degree of counter-ion dissociation was independent of carbon number of 1-alkanols. Comparing the results of the present system with those of sodium dodecylsulfate (SDS) micelle/1-alkanol system, it was found that the interior of DDAB vesicle membrane is more hydrophobic than that of SDS micelles. Furthermore, the increase in counter-ion dissociation caused by solubilized 1-alkanols was much lesser for DDAB vesicles than SDS micelles. PMID- 19282641 TI - Protonation behavior and stability of micelles of N-lauroylaminoalkyl dimethylamine oxides--effects of added salt concentration and spacer length. AB - The protonation behavior and the stability of micelles of N-lauroylaminopropyl N',N'- dimethylamine oxide (C12AmCn; n=3) were studied by the hydrogen ion titration and the equilibrium surface tension measurement. The surface potential of C12AmC3 micelles estimated from the results of hydrogen ion titration was lower than that of dodecyldimethylamine oxide micelles at added NaCl concentration, C(S), of 0.1M. The critical micelle concentration (cmc) estimated from surface tension measurements revealed that a reversal of the stability of the cationic and the nonionic micelles of C12AmC3 takes place in the range of C(S) higher than about 0.2M, that is, the cationic micelles become more stable than the nonionic ones in the range C(S) >or= 0.2M. In addition, the surface tension measurements were carried out for C12AmCn with different spacer length, n, ranging from 2 to 6 methylene units at C(S) = 0.1M. The cmc values of nonionic species were kept at almost constant for n or= 4. The cmc values of cationic species also tended to decrease with n for n >or= 4. These results were compared with those obtained for alkyldimethylamine oxides, and the effect of amide group and spacer groups introduced into amine oxide surfactants was discussed. PMID- 19282642 TI - Phase behavior and froth stability in a water/lysophospholipid system. AB - Phase behavior in a water/lysophospholipid system was investigated. A hydrated solid phase is observed at low temperature. Above the melting temperature of the hydrated solid a micellar phase at low surfactant concentration and a liquid crystalline phase at high surfactant concentration were observed. Similarly to poly(oxyethylene)-type nonionic surfactant systems, clouding behavior takes place at high temperature. The concentration of micelle-liquid crystal transition shifts to higher surfactant concentration in acidic or basic condition, but polyhydric alcohol addition does not affect very much on this transition point. The effect of added inorganic salts on the phase behavior was also studied, and the change in melting temperature of the hydrated solid and the cloud point shows a trend of the order of Hofmeister series. At room temperature a solid phase and an aqueous phase are equilibrated and one can obtain rather stable froth by shaking the sample that may be stabilized by the adsorption of solid dispersions on the air-water interface. The effect of inorganic salts on the froth stability in the water/lysophospholipid system was studied. The inorganic salts with salting-out effect improved froth stability. PMID- 19282643 TI - Fabrication and photocatalytic activity of TiO2/MoO3 particulate films. AB - TiO(2) and TiO(2)/MoO(3) particulate films were successfully prepared using a Langmuir-Blodgett technique. The as-deposited films were characterized by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and electron probe microanalysis (EPMA). The results indicate that the TiO(2) and TiO(2)/MoO(3) films are composed of separate layers of TiO(2) and MoO(3) particles. Furthermore, the photocatalytic activity of the films was evaluated from the decomposition of a stearic acid film under ultraviolet and visible light irradiation. It was found that photocatalytic activity is significantly enhanced by the layer structure of the TiO(2)/MoO(3) film, compared to the single layer TiO(2) film. PMID- 19282644 TI - Pharmacological activity of compounds extracted from persimmon peel (Diospyros kaki THUNB.). AB - Persimmon peels (Diospyros kaki THUNB.) are discarded during the production of dried fruit. The 2-methoxy-4-vinylphenol (8) which is component of persimmon peel had high antioxidant activity on the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay and SOD (superoxide dismutase) assay. And (8) had higher tyrosinase inhibiting activity than that of arbutin using both L-tyrosine and L DOPA as substrates. In addition, tyrosinase inhibiting activity of synthesized 2 methoxy-4-vinylphenol glycoside (8a) was studied. (8a) had tyrosinase inhibiting activity, suggesting that (8a) has possibilities for ingredient of cosmetics that are possessed of whitening effect. PMID- 19282645 TI - Raman tensors and their application in structural studies of biological systems. AB - The Raman scattering of a molecule is generated by interactions of its electrons with incident light. The electric vector of the Raman scattered light is related to the electric vector of the incident light through a characteristic Raman tensor. A unique Raman tensor exists for each Raman-active molecular vibrational mode. In the case of biologically important macromolecules Raman tensors have been determined for a few hundred vibrational Raman bands. These include proteins and their amino acid constituents, as well as nucleic acids (DNA and RNA) and their nucleotide constituents. In this review Raman tensors for 39 representative vibrational Raman bands of biological molecules are considered. We present details of the Raman tensor determinations and discuss their application in structural studies of filamentous bacteriophages (fd, Pf1, Pf3 and PH75), fowl feather rachis and eyespots of the protists, Chlamydomonas and Euglena. PMID- 19282646 TI - Stable delta(15)N and delta(13)C isotope ratios in aquatic ecosystems. AB - In the past 20 years, rapid progress in stable isotope (SI) studies has allowed scientists to observe natural ecosystems from entirely new perspectives. This report addresses the fundamental concepts underlying the use of the SI ratio. The unique characteristics of the SI ratio make it an interdisciplinary parameter that acts as a chemical fingerprint of biogenic substances and provides a key to the world of isotopomers. Variations in SI ratios of biogenic substances depend on the isotopic compositions of reactants, the pathways and kinetic modes of reaction dynamics, and the physicochemical conditions. In fact, every biogenic material has its own isotopic composition, its "dynamic SI fingerprint", which is governed by its function and position in the material flow. For example, the relative SI ratio in biota is determined by dietary lifestyle, e.g., the modes of drinking, eating, and excreting, and appears highly regular due to the physicochemical differences of isotopomers. Our primary goal here is to elucidate the general principals of isotope partitioning in major biophilic elements in molecules, biogenic materials, and ecosystems (Wada, E. et al., 1995). To this end, the nitrogen and carbon SI distribution ratios (delta(15)N and delta(13)C, respectively) are used to examine materials cycling, food web structures, and their variability in various kinds of watershed-including aquatic ecosystems to elucidate an "isotopically ordered world". PMID- 19282647 TI - Gene content, organization and molecular evolution of plant organellar genomes and sex chromosomes: insights from the case of the liverwort Marchantia polymorpha. AB - The complete nucleotide sequence of chloroplast DNA (121,025 base pairs, bp) from a liverwort, Marchantia polymorpha, has made clear the entire gene organization of the chloroplast genome. Quite a few genes encoding components of photosynthesis and protein synthesis machinery have been identified by comparative computer analysis. We also determined the complete nucleotide sequence of the liverwort mitochondrial DNA and deduced 96 possible genes in the sequence of 186,608 bp. The complete chloroplast genome encodes twenty introns (19 group II and 1 group I) in 18 different genes. One of the chloroplast group II introns separates a ribosomal protein gene in a trans-position. The mitochondrial genome contains thirty-two introns (25 group II and 7 group I) in the coding regions of 17 genes. From the evolutionary point of view, we describe the origin of organellar introns and give evidence for vertical and horizontal intron transfers and their intragenomic propagation. Furthermore, we describe the gene organization of the Y chromosome in the dioecious liverwort M. polymorpha, the first detailed view of a Y chromosome in a haploid organism. On the 10 megabase (Mb) Y chromosome, 64 genes are identified, 14 of which are detected only in the male genome. These 14 genes are expressed in reproductive organs but not in vegetative thalli, suggesting their participation in male reproductive functions. These findings indicate that the Y and X chromosomes share the same ancestral autosome and support the prediction that in a haploid organism essential genes on sex chromosomes are more likely to persist than in a diploid organism. PMID- 19282649 TI - [Health status of precarious workers in "Toshikoshi Haken Mura (Dispatch Workers' New Year Village)"]. AB - In Japan, the liquidity of employment is progressing rapidly. Due to the global recession which started in 2008, large-scale unemployment of precarious workers, including dispatch workers, is expected in 2009. In recent studies, it has been suggested that the unstable employment status itself may affect the health conditions of such workers. At the end of 2008, "Toshikoshi Haken Mura" (Dispatch Workers' New Year Village) was established in central Tokyo as a shelter for about 500 workers who had lost their jobs and accommodation. We participated in health consultations and medical checkups conducted by the Tokyo Metropolitan Government on January 8-10, 2009 to investigate the health conditions of the "villagers". Eighty-nine people attended health consultations and medical checkups (mean age, 48 yr). Physical complaints in the order of prevalence were as follows: respiratory (cough, 43%; sputum, 36%), low-grade fever (16.9%), musculoskeletal (13.5%), dermal (5.6%), digestive (3.4%), neurological (3.4%) and others including mental complaint (10.1%) such as anxiety, insomnia, and depression. During individual consultations, we found many cases who had had difficulty in getting medical consultations and whose treatment had been discontinued. Moreover, the rate of getting periodic medical examination remained low at 23.8% (among 84 people). Health problems of precarious workers have not been sufficiently recognized; however, there were cases whose poor health was indeed related to unstable employment. Studies on the health effects of temporary work should have higher prioritizes in the occupational health field. PMID- 19282648 TI - Discovery of alpha-Klotho unveiled new insights into calcium and phosphate homeostasis. AB - alpha-Klotho was first identified as the responsible gene in a mutant mouse line whose disruption results in a variety of premature aging-related phenotypes. alpha-Klotho has been shown to participate in the regulation of parathyroid hormone secretion and trans-epithelial transport of Ca(2+) in the choroid plexus and kidney. alpha-Klotho, acting as a cofactor for FGF23, is also a major regulator of vitamin D biosynthesis and phosphate reabsorption in the kidney. These suggest that alpha-Klotho is a key player that integrates a multi-step regulatory system of calcium and phosphate homeostasis. Collectively, the molecular function of alpha-Klotho reveals a new paradigm that may change current concepts in mineral homeostasis and give rise to new insights in this field. PMID- 19282650 TI - Validity and reliability of the Frenchay Activities Index for community-dwelling elderly in South Korea. AB - The speed of population aging in South Korea is projected as the fastest in the world, and a long-term care insurance law has been just enacted. Therefore, it is important to estimate the number of the disabled community-dwelling elderly in South Korea. The Frenchay Activities Index (FAI) is a scale to measure higher level of physical function or activities that are necessary for independence in the community, i.e., instrumental activities of daily living (IADL). The FAI has undergone the most intensive evaluation, but there are only a few studies to establish its validity and reliability for community-dwelling elderly. In addition, a Korean version of the FAI has not been developed yet. The purpose of this study was to develop a Korean version of the FAI and analyze the related factors with IADL, using data from 770 community-dwelling elderly people in Deajeon, Korea. In the results, the reliability was acceptable (Cronbach's alpha coefficient = 0.796). The content and discriminative validity were found to be satisfactory. In the results of the factor analysis, a reasonable four-component solution (domestic chores, outdoor work, leisure and hobby) was found. The related factors to IADL were age, gender, education level, and living conditions. The present findings suggest that the Korean version of the FAI would be useful as a measure of IADL and the results of the factor analysis might reflect socio cultural differences, such as illiteracy, low socioeconomic level, and sex roles of the Korean elderly. PMID- 19282651 TI - Decline of plasma brain natriuretic peptide during enzyme replacement therapy in a female patient with heterozygous Fabry's disease. AB - There are no data regarding changes in plasma brain natriuretic peptide (BNP) levels in patients with Fabry's diseases during enzyme replacement therapy (ERT). We describe a patient with Fabry's disease who demonstrated the improvement in plasma brain BNP levels in response to ERT. Fabry's disease is an X-linked lysosomal storage disorder caused by a deficiency of the enzyme alpha galactosidase A, which results in progressive intracellular accumulation of globotriaosylceramide (Gb3) in various organs including the heart. Cardiac involvement is frequent in Fabry's disease, resulting in cardiac dysfunction due to hypertrophic changes of the myocardium and thickening of the valves. Although ERT has been reported to improve cardiac function, no consensus has been reached regarding the effectiveness of ERT in female patients with heterozygous Fabry's disease. We report a 44-year-old woman having heterozygous Fabry's disease, who showed mitral valve thickening and regurgitation on echocardiogram. ERT was performed by intravenous infusion of recombinant alpha-galactosidase A every 2 weeks. We assessed the influences of ERT on cardiac function by measuring echocardiograhic parameters and monitoring BNP levels, which show treatment induced drop in patients with heart failure. Although her cardiac function and mitral regurgitation assessed by echocardiography had not improved 18 months after the beginning of ERT, the plasma BNP level, which was 91.5 pg/ml before ERT, fell to 18.9 pg/ml. In conclusion, plasma BNP levels may be useful for evaluating the effectiveness of ERT for heterozygous Fabry's disease, even in patients who demonstrate no improvement in echocardiographic parameters of cardiac structure and function. PMID- 19282652 TI - Toll-like receptor 3 signaling induces chronic pancreatitis through the Fas/Fas ligand-mediated cytotoxicity. AB - Innate immunity plays important roles in host defense against pathogens, but may also contribute to the development of autoimmune diseases under certain conditions. Toll-like receptors (TLRs) recognize various pathogens and induce innate immunity. We herein present a mouse model for chronic pancreatitis, which was induced by TLR3 signaling that generated the Fas/Fas ligand (FasL)-mediated cytotoxicity. An analogue of viral double-stranded RNA, polyinosinic:polycytidylic acid (poly I:C), which is recognized by TLR3, was injected into autoimmune-prone strains: MRL/Mp mice (MRL/+), MRL/Mp mice with a deficit of Fas (MRL/lpr) and MRL/Mp mice with a deficit of functional FasL (MRL/gld). The pancreatitis in MRL/+ mice was initiated by the destruction of pancreatic ductules, and its severity was significantly higher than that in MRL/lpr mice or MRL/gld mice. Using a pancreatic duct epithelial cell line MRL/S 1 newly established from the MRL/gld mouse that lacks FasL, we showed that treatment with poly I:C significantly induced the expression of Fas on the cultured cells. MRL/S-1 cells were destructed when co-cultured with splenocytes bearing intact FasL prepared from MRL/+ or MRL/lpr mice, but the magnitude of cytotoxicity was smaller with splenocytes of MRL/gld mice. Likewise, synthetic FasL protein showed cytotoxicity on MRL/S-1 cells. Furthermore, MRL/S-1 cells expressed higher levels of chemokines after the treatment with poly I:C, suggesting that the poly I:C-mediated induction of chemokines may be responsible for recruitment of lymphoid cells to the pancreatic periductular regions. These findings indicate that TLR3 signaling generates the Fas/FasL-mediated cytotoxicity, thereby leading to the development of chronic pancreatitis. PMID- 19282653 TI - Cysteinyl leukotrienes enhance the degranulation of bone marrow-derived mast cells through the autocrine mechanism. AB - The cysteinyl leukotrienes (LTs), LTC(4), LTD(4), and LTE(4), are potent inflammatory mediators and are involved in allergic reactions, such as bronchoconstriction, eosinophilic inflammation, and allergic cell proliferation. The present study aimed to elucidate the role of constitutively produced cysteinyl LTs in mast cell activation. We used a newly developed quantification method based on mass spectrometry to detect cysteinyl LTs in the cultured medium of mouse bone marrow-derived mast cells (BMMCs), which were obtained by interleukin (IL)-3-conditioned culture of mouse bone marrow. BMMCs were stimulated with immunoglobulin (Ig) E and antigen (IgE/Ag) or lipopolysaccharide for 1 or 24 h. This new quantification method revealed that unstimulated BMMCs produced and secreted LTB4 and LTE4 after 24 h of incubation. The treatment of unstimulated BMMCs for 2 h with montelukast, an antagonist of a cysteinyl LT receptor, CysLT1, resulted in the suppression of a downstream signaling event of this receptor, i.e., the decrease in phosphorylation of extracellular responsive kinases. Thus, cysteinyl LTs constitutively simulate BMMCs through the CysLT1 receptor in an autocrine manner. Treatment of BMMCs for 3 weeks with montelukast, which caused long-term inhibition of the autocrine cyteinyl LT-derived signal, significantly attenuated the IgE/Ag-dependent degranulation, as judged by the decrease in the release of beta-hexosaminidase, an enzyme contained in the granules, whereas the production of cytokines, such as IL-6 and tumor necrosis factor-alpha, were largely unaffected. In conclusion, an autocrine signal derived from constitutively produced cysteinyl LTs predisposes mast cells to the degranulation upon allergic stimulation. PMID- 19282654 TI - Tranilast, an anti-allergic drug, down-regulates the growth of cultured neurofibroma cells derived from neurofibromatosis type 1. AB - Neurofibromas are benign tumors that comprise primarily of Schwann cells and fibroblasts. Mast cells have been found scattered in the tumor tissue, and their role in promoting the proliferation of neurofibroma has been suggested. Tranilast (N-[3,4-dimethoxycinnamolyl]anthranilic acid) is an anti-allergic drug that inhibits release of the chemical mediators from mast cells and it used for the treatment of keloids and hypertrophic scars by its inhibition of growth-promoting transforming growth factor (TGF)-beta(1) from fibroblasts. We assumed that tranilast would suppress neurofibroma cell growth. In order to prove this hypothesis, we investigated the effectiveness of tranilast in inhibiting the tumor growth using a new cell culture system obtained from patients with neurofibromas. We called this culture system with the mixture of Schwann cells and fibroblasts "NF1 cells culture". Mast cells were differentiated from CD34(+) peripheral blood mononuclear cells of normal healthy subjects, and were co cultured with NF1 cells. Three days after tranilast (10 approximately 100 microM) added to the culture dishes, we counted viable cell numbers and measured the concentrations of TGF-beta(1), stem cell factor (SCF) and tryptase, which exists in the histamine granule, in the culture medium. Tranilast significantly suppressed proliferation of the NF1 cells and lowered the levels of TGF-beta(1), SCF and tryptase. These results suggest that tranilast retards tumor proliferation through not only suppression of cell growth factor, but also the inhibition of a chemical mediator released from mast cells. Thus, tranilast can be a potent therapeutic agent to inhibit the growth of neurofibromas. PMID- 19282655 TI - Over-expression of c-FLIP confers the resistance to TRAIL-induced apoptosis on gallbladder carcinoma. AB - Gallbladder carcinoma (GBC) is an aggressive malignancy with high mortality, mainly due to the reduced chance of curative resection and the resistance to chemotherapeutic drugs. Here, we showed that cellular Fas-associated death domain like interleukin-1 converting enzyme inhibitory protein (c-FLIP), an anti apoptotic protein, was over-expressed in the most of gallbladder carcinoma tissues, as judged by immunohistochemistry. Semi-quantitation was performed by determining the percentage of c-FLIP-positive cells: no positive cells (-), approximately 1% positive cells (+), approximately 30% positive cells (++), and >70% positive cells (+++). Out of the 35 tissue specimens of gallbladder carcinoma, positive c-FLIP expression was found in 26 samples (6/positive+++, 13/++, 7/+), whereas negative or weak c-FLIP staining was detected in normal (1/+, 9/-) and adenomatous (2/+, 8/-) gallbladder tissues. Then, we used a small interference RNA (siRNA), which can substantially down-regulate the expression levels of c-FLIP mRNA and protein in GBC-SD and SGC-996 human gallbladder carcinoma cells, as confirmed by real-time PCR and western blot analyses. Furthermore, the combined treatment with the c-FLIP siRNA and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) significantly induced apoptosis in gallbladder carcinoma cells, as judged by the increases in pyknosis, caspase-3/7 activities, and Annexin V-propidium iodide labeling, a marker for chromatin condensation. Thus, the siRNA-mediated down-regulation of c-FLIP profoundly enhances the sensitivity to TRAIL-induced apoptosis. In conclusion, c FLIP expression is up-regulated in gallbladder carcinoma and the down-regulation of c-FLIP sensitizes TRAIL-induced apoptosis. The present study provides a potent strategy for the treatment of gallbladder carcinoma by targeting the c-FLIP. PMID- 19282656 TI - Lectin activity of the coagulation factor VIII/von Willebrand complex. AB - The human coagulation factor VIII (FVIII) is essential in the intrinsic pathway of blood coagulation and circulates mainly as a non-covalently bound complex with the von Willebrand factor (VWF). This complex (FVIII/VWF) protects FVIII from degradation and cellular uptake, although no biological role has been identified yet for this complex. The FVIII/VWF complex was purified from a healthy donor's plasma by affinity chromatography on a Sepharose 4B-Concanavalin A column and was used to determine its capability to interact with erythrocytes and platelets. The purified FVIII/VWF complex at 6.0 and 12 microg/ml agglutinates rabbit and bovine erythrocytes, and showed negative agglutination with erythrocytes from other species including human ABO. Treatment of erythrocytes with Clostridium perfringens sialidase or trypsin increased four-fold the activity toward rabbit erythrocytes and positive agglutination for human A and B erythrocytes, suggesting the presence of FVIII/VWF-cryptic receptors in these erythrocytes. Goat, pig, or human O erythrocytes were not agglutinated even after enzymatic treatment. Fucose or N-acetyl-glucosamine (GlcNAc), at 10 mM, inhibited agglutinating activity of the complex with rabbit, human A and B erythrocytes, whereas galactose and N-acetyl-galactosamine, even at 200 mM, showed no effect on the complex activity. The FVIII/VWF complex, at 1.5 microg/200,000 platelets, significantly decreased platelet aggregation (p < 0.001) when compared with the effect of platelet-rich plasma; this effect was inhibited with 15 mM GlcNAc or fucose. ELISA assays on FVIII/VWF coated polystyrene plates confirmed specific binding to fucose- or biotinylated GlcNAc-dextran derivatives. We therefore propose that the FVIII/VWF complex possesses lectin activity. PMID- 19282657 TI - Accurate diagnosis of peripheral small cell lung cancer with computed tomography. AB - Small lesions are frequently detected in the lung with computed tomography (CT) in clinical practice. It is important to know the CT features of small-sized peripheral small cell lung cancer (SCLC) for early-stage diagnosis. We reviewed the CT findings of SCLC that presented as a solitary peripheral nodule without associated lymphadenopathy. This study included 12 patients (11 men and 1 woman; mean age, 68.5 years) with peripheral SCLC of diameters ranging from 9 - 28 mm (mean, 15.4 mm). We evaluated the findings with thin-section CT for each peripheral tumor; emphasis was laid on the predominant internal characteristics (whether the mass is solid), tumor-lung interface characteristics (whether the mass is well-defined with a smooth surface or with lobulation or spiculation), and surrounding structures (the presence or absence of perivascular thickening adjacent to the tumor). In all patients, most portions of the tumor consisted of a non-calcified solid mass. Contrast enhancement in varying degrees was observed in the tumors of all 8 patients, who were evaluated with enhanced CT. The tumor lung interface characteristics observed on the CT images included a well-defined mass with a smooth surface (n = 5), a well-defined mass with lobulation (n = 3), and a mass with spiculation (n = 4). An irregular perivascular thickening adjacent to the tumor was observed in 4 patients. We conclude that peripheral SCLC without associated lymphadenopathy manifests as a non-calcified solid mass and is occasionally characterized by perivascular thickening. PMID- 19282658 TI - Bach1 deficiency ameliorates hepatic injury in a mouse model. AB - Bach1 is a basic region-leucine zipper (bZip) protein that forms heterodimers with the small Maf proteins and functions as a repressor of gene expression. One of the target genes of Bach1 is Hmox-1 that encodes heme oxygenase-1 (HO-1). HO-1 degrades heme into carbon monoxide (CO), biliverdin, and iron. HO-1 is strongly induced by various stresses as well as its substrate heme, and protects cells and tissues against insults through diverse cytoprotective functions of the reaction products CO and biliverdin. Bach1-deficiency in mice leads to higher expression of Hmox-1 in various tissues. Here we investigated the effects of Bach1 deficiency in mice on tissue injuries: hepatic injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS), and mouse paw edema induced by carrageenin, polysaccharide derived from various seaweeds. Bach1-deficiency suppressed induction of plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in response to the GalN/LPS-treatment. However, production of tumor necrosis factor alpha (TNF-alpha) and nitric oxide (NO), both being cytotoxic mediators in LPS-induced hepatic injury, in Bach1-deficient mice and their peritoneal macrophages was similar to wild type controls. In contrast, Bach1-deficiency did not affect extent of mouse paw edema induced by carrageenin, which enhances vascular permeability by activating kinin release. These results indicate that Bach1 plays an inhibitory role in the cytoprotection of LPS-induced liver injury but not in the kinin-mediated inflammatory edema. The inhibitory role for Bach1 may stem from its activity to repress gene expression including HO 1. PMID- 19282659 TI - Establishment of a screening system for chemicals that upregulate a melanoma antigen, Melan-A/MART-1. AB - Immunotherapy is well-practiced as one of the main adjuvant therapies for melanoma patients. Until now, many immunotherapeutic investigations have focused on improving the effector side of the antitumor response, but only a few studies have been concerned with preventing the loss of tumor-associated antigen (TAA) expression. Loss of TAA should be an important problem for the recognition of tumor cells by cytotoxic T lymphocytes. If any agents that augment the expression of melanoma antigens were found, they could improve the efficacy of immunotherapy by increasing the antigens. To detect effective chemicals, we made a fluorescent cellular reporter system for screening promising candidate chemicals. In this system, the fusion gene of the Melan-A/MART-1 promoter sequence followed by the green fluorescent protein (GFP) coding region was stably transfected into MUX human melanoma cells which are known to express little or no Melan-A/MART-1. Melan-A/MART-1 is a well-known melanoma antigen recognized by autologous cytotoxic T cells, and is a glycoprotein associated with the melanosome, the organelle in which melanin synthesis proceeds. By using this screening system, daunorubicin, doxorubicin and cytochalasin D, which enhanced the green fluorescent, were selected and then were confirmed to actually increase the expression of Melan-A/MART-1 mRNA and protein in human melanoma cells of MU89, MM96L(+) and SK-MEL-28, but also in low-antigen presenting cells such as MM96L( ), MUX, and A375. In conclusion, we have successfully established a well functioning screening system, which will allow us to find candidate chemicals that up-regulate or maintain the melanoma antigen expression. PMID- 19282660 TI - Restraint stress impairs erectile responses in rats. AB - It has been established that various forms of physical and psychological stress reduce sexual functions. However, there is no study yet evaluating the functional changes over cavernosal pressure in rats exposed to restraint stress. In this study, we aimed to investigate the convenience of the restraint stress model that may be used to determine the disruptive effects of stress on erectile function. Sprague Dawley rats were randomized into two groups as control (n = 7) and stress (n = 7) groups. In the stress group, rats were placed for 60 minutes in a cylindrical plastic tube with holes for fresh air supply (restraint stress). Following the stress application, several parameters for erectile responses were evaluated immediately. The control animals were maintained at room temperature without any procedure until the measurement. During the electrical stimulation of cavernous nerve, we measured the intracavernous pressure (ICP), the ratio of ICP to the mean arterial pressure (MAP), and detumescence time. There were significant decreases in ICP (24.4 +/- 4.1 vs 53.4 +/- 4.5 mmHg, p < 0.01), ICP/MAP (34.4 +/- 7.8% vs 55.7 +/- 3.9%, p < 0.05), and detumescence time (31.7 +/- 6.1 vs 78.6 +/- 12.8 sec, p < 0.01) in stress group when compared to control group. Thus, restraint stress declined detumescence time and decreased intracavernosal pressure in male rats. In conclusion, restraint stress model in rats may be useful for determining the effects of stress on erectile response. Even a short-term restraint stress may cause erectile dysfunction. PMID- 19282661 TI - The benefits of supplementary private health insurance for healthcare utilization and survival among stomach cancer patients. AB - Social health insurance (SHI) has covered only 60% of healthcare payments in Korea; therefore, Koreans rely upon supplementary private health insurance (SPHI) to cover the rest of the payments. SPHI status is assumed to affect clinical treatment and outcome of patients with stomach cancer, which is the most prevalent cancer among Koreans. This study examined the relationships between SPHI and diagnosis, treatment, utilization, and survival among stomach cancer patients. Patients (n = 3,780) who underwent a radical gastrectomy from 2000 to 2003 were examined retrospectively. The mean age of all patients was 58 years, and 1,377 patients (36%) had one or more SPHI plan. Univariate analysis was used to examine differences between SPHI and non-SPHI patients in terms of demographic, socioeconomic, and clinical characteristics, followed by a multiple analysis with adjustment for the above parameters. SPHI patients were 5% more likely to visit a physician (p < 0.001) and were 0.8 times less likely to die (p = 0.03) than non-SPHI patients. In contrast, no difference was observed between the two groups in the stage of cancer at diagnosis, which may reflect the fact that the majority of Korean SPHI plans do not cover cancer screening services. Moreover, no significant difference was detected in the use of adjuvant therapy between groups, because it was covered by SHI. In conclusion, SPHI is a determinant of healthcare utilization and survival among stomach cancer patients. It is necessary for health policymakers to consider the benefits of SPHI in financing healthcare services. PMID- 19282662 TI - Reprogramming cell fates in the mammary microenvironment. AB - The capacity of any portion of the murine mammary gland to produce a complete functional mammary outgrowth upon transplantation to an epithelium-divested fat pad is unaffected by the age or reproductive history of the donor. Likewise, through serial transplantations, no loss of potency is detected when compared to similar transplantations of the youngest mammary tissue tested. This demonstrates that stem cell activity is maintained intact throughout the lifetime of the animal despite aging and the repeated expansion and depletion of the mammary epithelium through multiple rounds of pregnancy, lactation and involution. These facts support the contention that mammary stem cells reside in protected tissue locales (niches), where their reproductive potency remains essentially unchanged through life. Disruption of the tissue, to produce dispersed cells results in the desecration of the protection afforded by the "niche" and leads to a reduced capacity of dispersed epithelial cells (in terms of the number transplanted) to recapitulate complete functional mammary structures. Our studies demonstrate that during the reformation of mammary stem cell niches by dispersed epithelial cells in the context of the intact epithelium-free mammary stroma, non-mammary cells may be sequestered and reprogrammed to perform mammary epithelial cell functions including those ascribed to mammary stem/progenitor cells. PMID- 19282663 TI - FANCM-FAAP24 and HCLK2: roles in ATR signalling and the Fanconi anemia pathway. AB - The ATR signalling pathway coordinates the cellular response to replication stress, which is essential for the maintenance of genome integrity. HCLK2/Tel2 is a highly conserved orphan protein that binds directly to ATR and other PI3-kinase related kinases and plays a central role in checkpoint signalling responses.(1) Proteomic analyses of HCLK2 complexes confirmed ATR, ATRIP and DNA-PKcs as HCLK2 interacting factors and also uncovered two surprising interacting proteins, the heterodimeric Fanconi Anemia (FA) proteins FANCM and FAAP24. Our subsequent findings that ATR signalling is attenuated in FANCM and FAAP24-depleted cells, together with recent biochemical studies, suggested that remodelling of stalled replication forks by FANCM-FAAP24 is required to facilitate efficient activation of ATR signalling in response to replication stress.(2) Furthermore, our study revealed that the DNA translocase activity of FANCM is essential for efficient activation of the ATR signalling, a function that is separate and distinct from its role in targeting the FA core complex to sites of DNA damage. In this review we discuss the importance of these findings in the context of recent data and raise questions regarding the role of HCLK2 and FANCM-FAAP24 in human disease. PMID- 19282664 TI - BubR1 is an effector of multiple mitotic kinases that specifies kinetochore: microtubule attachments and checkpoint. AB - BubR1 is a critical component of the mitotic checkpoint but has also been shown to play an essential role in establishing kinetochore:microtubule attachments. BubR1 is hyperphosphorylated in mitosis and recent studies in human and Xenopus have identified 9 phosphorylation sites. Plk1-dependent phosphorylations (T792, T1008 and S676) were reported to stimulate BubR1 kinase activity, promote kinetochore microtubule attachments, monitor kinetochore tension, as well as the recruitment of Mad2 checkpoint protein to kinetochores. Plk1-independent sites (S435, S543, S670 and S1043) were also identified and some of these were found to be sensitive to the loss of microtubule attachment but not tension. Functional studies showed that phosphorylation of S670 is critical for correcting aberrant attachments. Once end-on attachments are established, dephosphorylation of S670 appeared to be important for generating tension to signal anaphase onset. The collective data when combined with early EM studies that showed BubR1 is present at both the inner and outer kinetochore plates suggest that BubR1 maybe an effector of multiple kinases that specifies its roles in microtubule attachments and checkpoint functions. PMID- 19282665 TI - Homeodomain protein Dlx3 induces phosphorylation-dependent p63 degradation. AB - The epidermis is a stratified epithelium which develops depending on the transcription factor p63, a member of the p53 family of transcription factors. p63 is strongly expressed in the innermost basal layer where highly proliferative epithelial cells reside. p63 functions as a molecular switch that initiates epithelial stratification or cell fate determination while regulating proliferation and differentiation of developmentally mature keratinocytes. p63 acts upstream of Dlx3 homeobox gene in a transcriptional regulatory pathway relevant to ectodermal dysplasia. Here we show that Dlx3 triggers p63 protein degradation by a proteasome-dependent pathway. Mutant DeltaNp63alpha in which Threonine397 and Serine383 were replaced with Alanine as well as C-terminal truncated versions of DeltaNp63alpha are resistant to Dlx3-mediated degradation. Transient expression of Dlx3 is associated with Raf1 phosphorylation. Dlx3 is unable to promote p63 degradation in Raf1 depleted MEF cells or upon pharmacological knockdown of Raf1. Our data support a previously unrecognized role for Dlx3 in posttranslational regulation of DeltaNp63alpha protein level, a mechanism that may contribute to reduce the abundance of DeltaNp63alpha during differentiation of stratified epithelia. PMID- 19282666 TI - A new take on ceramide: starving cells by cutting off the nutrient supply. AB - Ceramide generation is increased by a broad array of signals. In general, ceramide limits cell survival and proliferation and promotes differentiation and senescence. Despite its role in the pathogenesis of multiple human diseases, ceramide's mechanism of action remains poorly defined. Understanding how this sphingolipid modulates cell physiology is therefore an important goal. Building on prior observations that ceramide induces autophagy, we demonstrate that ceramide kills cells by inducing severe bioenergetic stress secondary to nutrient transporter downregulation. In support of this model, maintaining nutrient access blocks ceramide-induced autophagy and cell death. This bioenergetic mechanism of action may explain the increased sensitivity of cancer cells to ceramide. Starvation induces quiescence in normal cells. Tumor cells, in contrast, carry oncogenic mutations that block the switch to catabolism and prevent a reduction in metabolic demand leading to a bioenergetic crisis when nutrients become scarce. We propose that the non-lethal effects of ceramide might also stem from ceramide-induced starvation. While severe nutrient stress kills cells, mild nutrient limitation slows proliferation and may contribute to the induction of senescence. In sum, our new model for ceramide action suggests that regulated nutrient transporter expression may play a previously unappreciated role in cancer and other diseases where ceramide metabolism is altered. PMID- 19282667 TI - SIRT2 downregulation confers resistance to microtubule inhibitors by prolonging chronic mitotic arrest. AB - We previously identified SIRT2, a deacetylase for tubulin and histone H4, as a protein downregulated in gliomas, and reported that exogenously-expressed SIRT2 arrests the cell cycle prior to entry into mitosis to prevent chromosomal instability in response to microtubule inhibitors (MTIs) such as nocodazole, characteristics previously reported for the CHFR protein. We herein investigated the effects of SIRT2 downregulation on sensitivity to MTIs using HCT116 cells, a mitotic checkpoint-proficient near-diploid cancer cell line used for studying checkpoints. We found that SIRT2 downregulation confers resistance to MTIs as well as that of BubR1, a well-characterized mitotic checkpoint protein, though by a different mechanism. While BubR1 suppression abolished spindle checkpoint functions, which is a requirement for cell death after release from the spindle checkpoint, SIRT2 downregulation prolonged chronic mitotic arrest from sustained activation of the mitotic checkpoint and consequently prevented a shift to secondary outcomes, including cell death, after release from chronic mitotic arrest. Consistent with this notion, BubR1 downregulation was dominant over SIRT2 knockdown in regard to mitotic regulation in the presence of nocodazole. These results suggest that SIRT2 functions to release chronic mitotic arrest in cells treated with MTIs, leading to other outcomes. We also found that SIRT2 downregulation caused centrosome fragmentation in response to nocodazole prior to the alteration in spindle checkpoint function, implying not only a novel function of SIRT2 for centrosome maintenance upon exposure to mitotic stress caused by MTIs, but also the existence of a centrosome-mediated signaling pathway to sustain the spindle checkpoint. Therefore, this study highlights a novel pathway leading to resistance to MTIs, in which SIRT2 downregulation participates. PMID- 19282668 TI - Inhibition of cell differentiation: a critical mechanism for MYC-mediated carcinogenesis? AB - Despite its early discovery and relevance in cancer, the mechanisms by which MYC brings about tumorigenic transformation have not been clarified. MYC elicits a variety of biological activities, the proliferation promotion being the best studied. However, inhibition of cell differentiation was one of the first MYC activities described. The importance of differentiation impairment in MYC-induced tumorigenesis is demonstrated in transgenic mice models with conditional MYC expression, where MYC inactivation leads to tumor regression associated to re differentiation of tumor cells. To explain the anti-differentiation effects of MYC, it has been argued that MYC impairs differentiation by preventing exit from the cell cycle. However, we have described models where MYC blocks the erythroid or neuronal differentiation without reversing the proliferative arrest. In these and other models discussed here, MYC-mediated inhibition of differentiation occurs by blocking the upregulation of transcription factors that control the differentiation. The importance of MYC anti-differentiation function has recently gained importance after the discovery that MYC is one of the four transcription factors able to reprogram differentiated cells into pluripotent cells. We will discuss the hypothesis that MYC engages common pathways as a "stemness" keeper and as an oncogene. PMID- 19282669 TI - How ERK1/2 activation controls cell proliferation and cell death: Is subcellular localization the answer? AB - Extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) are members of the mitogen-activated protein kinase super family that can mediate cell proliferation and apoptosis. The Ras-Raf-MEK-ERK signaling cascade controlling cell proliferation has been well studied but the mechanisms involved in ERK1/2 mediated cell death are largely unknown. This review focuses on recent papers that define ERK1/2 translocation to the nucleus and the proteins involved in the cytosolic retention of activated ERK1/2. Cytosolic retention of ERK1/2 denies access to the transcription factor substrates that are responsible for the mitogenic response. In addition, cytosolic ERK1/2, besides inhibiting survival and proliferative signals in the nucleus, potentiates the catalytic activity of some proapoptotic proteins such as DAP kinase in the cytoplasm. Studies that further define the function of cytosolic ERK1/2 and its cytosolic substrates that enhance cell death will be essential to harness this pathway for developing effective treatments for cancer and chronic inflammatory diseases. PMID- 19282670 TI - Chk1 phosphorylation during mitosis: a new role for a master regulator. AB - The human DNA damage responses are modulated by both nonessential and essential pathways. The extensively studied ATM kinase and p53 are examples of the former. While loss-of-function mutations in genes that encode ATM and p53 cause marked predispositions to cancer, the loss of these proteins does not appear to impact basic cell growth and proliferation. In contrast, the checkpoint kinase Chk1 and its upstream activator ATR are essential.(1-4) What do these proteins do in undamaged cells? PMID- 19282671 TI - How does a millimeter-sized cell find its center? AB - Microtubules play a central role in centering the nucleus or mitotic spindle in eukaryotic cells. However, despite common use of microtubules for centering, physical mechanisms can vary greatly, and depend on cell size and cell type. In the small fission yeast cells, the nucleus can be centered by pushing forces that are generated when growing microtubules hit the cell boundary. This mechanism may not be possible in larger cells, because the compressive force that microtubules can sustain are limited by buckling, so maximal force decreases with microtubule length. In a well-studied intermediate sized cell, the C. elegans fertilized egg, centrosomes are centered by cortex-attached motors that pull on microtubules. This mechanism is widely assumed to be general for larger cells. However, re evaluation of classic experiments in a very large cell, the fertilized amphibian egg, argues against such generality. In these large eggs, movement of asters away from a part of the cell boundary that they are touching cannot be mediated by cortical pulling, because the astral microtubules are too short to reach the opposite cell boundary. Additionally, Herlant and Brachet discovered a century ago that multiple asters within a single egg center relative to the cell boundary, but also relative to each other. Here, we summarize current understanding of microtubule organization during the first cell cycle in a fertilized Xenopus egg, discuss how microtubule asters move towards the center of this very large cell, and how multiple asters shape and position themselves relative to each other. PMID- 19282673 TI - Creating a culture of discovery through clinical trials and translational research. PMID- 19282672 TI - CDK5RAP2 is required for spindle checkpoint function. AB - The combination of paclitaxel and doxorubicin is among the most successful chemotherapy regimens in cancer treatment. CDK5RAP2, when mutated, causes primary microcephaly. We show here that inhibition of CDK5RAP2 expression causes chromosome mis-segregation, fails to maintain the spindle checkpoint, and is associated with reduced expression of the spindle checkpoint proteins BUBR1 and MAD2 and an increase in chromatin-associated CDC20. CDK5RAP2 resides on the BUBR1 and MAD2 promoters and regulates their transcription. Furthermore, CDK5RAP2 knockdown cells have increased resistance to paclitaxel and doxorubicin, and this resistance is partially rescued upon restoration of CDK5RAP2 expression. Cancer cells cultured in the presence of paclitaxel or doxorubicin exhibit dramatically decreased CDK5RAP2 levels. These results suggest that CDK5RAP2 is required for spindle checkpoint function and is a common target in paclitaxel and doxorubicin resistance. PMID- 19282674 TI - Correlation of an audible fourth heart sound with level of diastolic dysfunction. AB - BACKGROUND: Expert physicians disagree on the usefulness of the fourth heart sound (S4) as an indicator of left ventricular (LV) diastolic dysfunction. METHODS: We correlated prevalence of the S4 with level of severity of LV diastolic dysfunction. From 551 consecutive echocardiography and Doppler studies, 106 patients in sinus rhythm but without cardiac conduction abnormalities, prosthetic or abnormal valves, or high blood flow states were auscultated by 3 investigators blinded to the participants' diastolic function as determined by 3 other cardiologists using mitral inflow, tissue, and pulmonary vein Doppler interrogation. RESULTS: Diastolic function was normal in 46 participants, 45 had mild, 10 had moderate, and 5 had severe diastolic dysfunction. S4 was audible in 35% with normal function, 42% with mild, 70% with moderate, and none with severe dysfunction (P = 0.052). Sensitivity was 43%, specificity 65%, and accuracy of 53% for discriminating normal from abnormal function. S4 is neither sensitive nor specific, is common but not normal in the elderly, may be absent with severe diastolic dysfunction, and therefore is not a useful indicator of LV diastolic dysfunction. CONCLUSIONS: The S4 is not an accurate indicator of diastolic dysfunction, as it is present in many persons with normal LV function and is absent in those with severe LV dysfunction. PMID- 19282675 TI - Serum ferritin levels predict all-cause and infection-cause 1-year mortality in diabetic patients on maintenance hemodialysis. AB - BACKGROUND: The aim of this study was to assess the relationship between the serum ferritin level and the 1-year outcome in diabetic maintenance hemodialysis (MHD) patients. METHODS: The prospective clinical study enrolled 187 diabetic MHD patients from a university hospital in Taiwan. All the patients were divided into 3 groups according to their serum ferritin levels: group I (<200 ng/mL; n = 71), group II (200-700 ng/mL; n = 97), and group III (>700 ng/mL; n = 19). A total of 26 demographic, clinical, and laboratory variables were analyzed as predictors of the 1-year mortality. RESULTS: There were no significant differences between these 3 groups except in their erythropoietin usage, hemoglobin, transferrin saturation, and high-sensitive C-reactive protein levels. The 1-year mortality rates were 9.2%, 11.4%, and 46.2% in groups I, II, and III, respectively. Group I and group II patients had a lower 1-year mortality rate than group III patients (log-rank test; chi = 8.807; P = 0.0112). CONCLUSION: The study suggested that serum ferritin levels predict both all-cause and infection-cause 1-year mortality in diabetic patients on MHD. In such patients, the serum ferritin levels are associated with both iron stores and the inflammation status. PMID- 19282676 TI - The hidden patient: addressing the caregiver. AB - Premium caregiving is pivotal to achieve quality care for persons with chronic or disabling conditions. Unpaid informal caregivers currently provide for a substantial proportion of care recipient needs including 80% of long-term care needs in the United States. Within the next 2 decades, 78 million baby boomers will reach 65 years of age and older, creating unique and extensive challenges to an already flawed health care system; this older cohort will at minimum increase utilization of health care resources. On an individual level, care providers need to support and educate caregivers and make caregivers partners in the delivery of excellent care. To achieve this, health care professionals need to educate themselves regarding the caregivers' demographics, ethnicity, characteristics, effective interventions, and outcomes. From a health policy perspective, public, private, and community funding needs to be allocated to advocate for, encourage, and train these caregivers. PMID- 19282677 TI - Giant emphysematous bulla mimicking tension pneumothorax. PMID- 19282679 TI - Suicidal behavior in schizophrenia: a test of the demoralization hypothesis. AB - This study examined Drake's model that individuals with schizophrenia with good premorbid adjustment and insight into their illness are more vulnerable to becoming demoralized and therefore suicidal. One hundred sixty-four patients with schizophrenia (N = 115) or schizoaffective disorder (N = 49) were assessed for depressive symptoms and DSM-III-R depression, premorbid functioning, insight and suicidal behavior using The Diagnostic Interview for Genetic Studies and the Premorbid Adjustment Scale. Premorbid adjustment, insight and past MDE did not discriminate attempters from nonattempters, contrary to the model. However, consistent with the model, the interaction between good premorbid adjustment and insight predicted severity of depressive symptoms, and the psychological symptoms of depression significantly differentiated attempters from nonattempters, whereas the somatic symptoms did not. This study provides support for some aspects of the demoralization model. PMID- 19282680 TI - Stressful life events, chronic difficulties, and the symptoms of clinical depression. AB - Major life events and chronic difficulties have been found to be associated with the onset of depression. Little is known, however, about how exposure to such stressors is related to the clinical presentation of this disorder. We addressed this issue by administering an interview-based measure of life stress, the Beck Depression Inventory, and the Global Assessment of Functioning scale to 100 adults diagnosed with major depressive disorder. Participants who experienced a preonset severe life event exhibited greater overall levels of depression severity, endorsed more cognitive and somatic symptoms of depression, and functioned at lower levels than did their counterparts without preonset severe life events. In contrast, exposure to a preonset severe difficulty was unrelated to participants' severity of depression, cognitive and somatic symptoms, or level of global functioning. These findings highlight the potentially greater importance of acute stress compared with chronic stress for influencing these key clinical features of depression. PMID- 19282681 TI - The factor structure of clinical symptoms in depressed inpatients with unipolar or bipolar spectrum disorders: a preliminary study. AB - Factor analysis of symptom structure has proven to be a valuable tool to identify dimensions of symptoms in various psychiatric conditions. This study used exploratory and confirmatory factor analysis to examine symptom structures in depressed inpatients with unipolar (n = 718) or bipolar (n = 134) spectrum disorders who were rated at admission with a psychiatric rating scale. No differences in overall symptom severity on the scale were found in the 2 samples, although different factor structures were detected with exploratory analyses. These models were modified in a confirmatory modeling procedure to improve their fit to the data, resulting in models with good, but not perfect, fits. For people with bipolar disorders, a 5-factor model fit best, with depression loading with anxiety symptoms and in people with unipolar disorders, a 4-factor model with depression loading with vegetative symptoms was found. Our results suggest that similar levels of symptom severity may have different underpinnings in the 2 groups and suggest that more comparative studies of symptoms in these 2 conditions may be useful. PMID- 19282682 TI - A pilot study of self-esteem as a mediator between family factors and depressive symptoms in young adult university students. AB - The aim of this study was to examine whether self-esteem mediates the relationship between family factors and depressive symptoms in young adults. Participants completed self-report questionnaires about overall family environment, conflict with mother or father, parental rearing, self esteem, and depressive symptoms. Self-esteem was found to mediate the relationship between the combined family factors and depressive symptoms. When examined simultaneously, none of the individual family variables uniquely predicted depressive symptoms or self-esteem. However, separate analysis of each of the three family factors provided evidence for self-esteem mediating the relationship between parental conflict and depressive symptoms, and the relationship between parental care and depressive symptoms. Self-esteem may play a role in the mechanism underlying the link between parent-offspring relationship factors and depressive symptoms. PMID- 19282683 TI - Predictive value of self-stigma, insight, and perceived adverse effects of medication for the clinical outcomes in patients with depressive disorders. AB - The aims of this prospective study were to examine the predictive values of self stigma, insight, and perceived adverse effects of medication for remission of depressive symptoms, suicidal risk, and medication adherence in patients with depressive disorders over a 1-year follow-up period. One hundred seventy-four participants who were in a state of obvious depression underwent an index interview to determine their degree of self-stigma, insight, and perceived adverse effects of medication. One year later, they were reassessed to determine the severity of their depressive symptoms, suicidal risk, and the level of the medication adherence, and their associations with the 3 possible predictors at the index interview were examined. The results of this study indicated that perceiving more severe adverse effects of medication at the index interview increased the risks of the nonremission of depressive symptoms, occurrence of suicidal ideation or attempt, and medication nonadherence in patients with depressive disorders in the 1-year period. However, the degrees of self-stigma and insight did not predict the severity of depressive symptoms, suicidal risk, or the level of the medication adherence. Based on the results of this study, we suggest that it is important for clinicians to prevent the occurrence of and to help patients manage the adverse effects of medication. We also suggest that further prospective studies are needed to examine the predictive values of self stigma and insight for clinical outcomes and medication adherence. PMID- 19282684 TI - Early stage assessment and course of acute stress disorder after mild traumatic brain injury. AB - Although it has been established that acute stress disorder (ASD) and posttraumatic stress disorder occur after mild traumatic brain injury (MTBI) the qualitative differences in symptom presentation between injury survivors with and without a MTBI have not been explored in depth. This study aimed to compare the ASD and posttraumatic stress disorder symptom presentation of injury survivors with and without MTBI. One thousand one hundred sixteen participants between the ages of 17 to 65 years (mean age: 38.97 years, SD: 14.23) were assessed in the acute hospital after a traumatic injury. Four hundred seventy-five individuals met the criteria for MTBI. Results showed a trend toward higher levels of ASD in the MTBI group compared with the non-MTBI group. Those with a MTBI and ASD had longer hospital admissions and higher levels of distress associated with their symptoms. Although many of the ASD symptoms that the MTBI group scored significantly higher were also part of a postconcussive syndrome, higher levels of avoidance symptoms may suggest that this group is at risk for longer term poor psychological adjustment. Mild TBI patients may represent a injury group at risk for poor psychological adjustment after traumatic injury. PMID- 19282685 TI - Mental health in adults with mild and moderate intellectual disabilities: the role of recent life events and traumatic experiences across the life span. AB - The aim of the present study is to investigate the association between recent life events and traumatic experiences across the life span and psychiatric disorders in people with ID. One hundred seventy-seven individuals with mild and moderate intellectual disability and their principal carers were assessed. Psychiatric disorders were evaluated with a semistructured psychiatric interview, the Psychiatric Assessment for Adults with Developmental Disabilities. This interview also includes a checklist of life events experienced over the previous 12 months, which was assessed through key informants. Presence of traumas was assessed through Allen's trauma history screen, also administered to key informants. After a descriptive analysis, binary logistic regression was used to see whether traumatic events and life events predicted the presence of ICD-10 disorders. A 75% of the participants had experienced at least 1 traumatic event during their life span, and 50% of the participants had experienced at least 1 life event in the 12 months previous to the study. Binary logistic regression showed that exposure to 1 or more traumatic experiences significantly increased the odds of a mental disorder (OR = 1.8), as did exposure to life events (OR = 1.4). However, when both life events and traumatic experiences were entered together in the model, calculation of odds ratios revealed that traumatic experiences significantly increased the odds of ICD-10 disorders (OR = 1.7) although life events were no longer significant. Though they have been less studied by the literature regarding predictors of mental illness in people with intellectual disability, traumatic experiences seem to play a more important role in psychopathology than life events. PMID- 19282686 TI - Nineteen-month stability of Revised NEO Personality Inventory domain and facet scores in patients with personality disorders. AB - We lack knowledge of the temporal stability of major personality dimensions in patients with personality disorders (PDs). The Revised NEO Personality Inventory (NEO-PI-R) is a self-report instrument that operationalizes the Five-Factor Model of personality. This study investigated the relative stability, mean level stability, and individual level stability of the NEO-PI-R scores in patients with PDs (n = 393) and patients with symptom disorders only (n = 131). The NEO-PI-R was administered at admission to short-term day treatment and after an average of 19 months. The results showed a moderate to high degree of stability of NEO-PI-R scale scores with no substantial difference in stability between patients with and without PD. Changes in NEO-PI-R scores were associated with changes in symptom distress. Neuroticism was the least stable domain. The study indicates that the Five-Factor Model of personality dimensions and traits are fairly stable in patients with PDs. The lower stability of Neuroticism may partly be explained by its inherent state aspects. PMID- 19282687 TI - Diagnosis of ovarian cancer and object relationships in early memories: a pilot study. AB - Women newly diagnosed with ovarian cancer (N = 16) and women from the community (N = 17) each described 3 early memories. We assessed the developmental maturity level of the memories with Mayman's Libidinal Level of Relationships scale, which assesses the maturity of memories form oral to latency levels. The early memories of the cancer patients were at a significantly lower libidinal level and more often involved oral relationship themes than the memories of the community women. The results suggest that a diagnosis of a life-threatening illness can invoke comforting memories of being cared for and fed, associated with a greater sense of dependence on others. PMID- 19282688 TI - Increased body mass index associated with increased harm avoidance and decreased self-directedness in Japanese women. AB - Previous studies have suggested that body mass index (BMI) is related to personality traits, and that there may be gender specificity in this relationship. In the present study, the association between BMI and the 7 dimensions of the Temperament and Character Inventory was investigated in 567 Japanese healthy volunteers, with special attention on gender effects. Height and weight were self-reported, and BMI was calculated from these values. In the multiple regression analyses, higher BMI was related to higher scores of harm avoidance (p < 0.05) and lower scores of self-directedness (p < 0.01) in women, whereas BMI was not related to any Temperament and Character Inventory dimension in men. The present study suggests that increasing BMI is associated with increased harm avoidance and decreased self-directedness in women but not in men in healthy subjects. PMID- 19282689 TI - Concern over weight and dieting in Taiwanese adolescents: correlations of gender, age, weight, and multidimensional characteristics. AB - The aims of this study were to compare the levels of concern over weight and dieting (COWD) among adolescents with different body weight status, gender, and age, and to examine the associations of COWD with multidimensional characteristics. Nine thousand eighty-seven adolescents completed the questionnaires. The levels of COWD were compared among the groups with different body weight status, gender, and age. The correlates of COWD in gender- and age specified groups were examined. The results indicated that the levels of COWD varied across the groups of adolescents with different body weight status, gender, and age. Being overweight or obese, depression, and low self-esteem were associated with high COWD in all gender- and age-specified groups. A variety of socio-demographic, personal, family, peer, and school characteristics were also associated with high COWD in adolescents with specified gender and age. Gender- and age-specified intervention programs targeting adolescents with the correlates of high COWD should be considered. PMID- 19282690 TI - Beliefs in traditional Chinese medicine efficacy among Chinese Americans: implications for mental health service utilization. AB - We examined how community attitudes towards traditional Chinese conceptions of health and Western dichotomization of illness might affect perceptions of traditional Chinese medicine (TCM) efficacy in order to identify factors underlying psychiatric service underutilization among Chinese-Americans. We administered an experimental vignette to assess perceived illness, severity, and beliefs of TCM efficacy for physical and psychiatric disorders among 90 Chinese Americans ascertained through a national telephone survey. Perceived illness severity was unrelated to assessment of TCM effectiveness. However, psychiatric conditions tended to be viewed as distinct from physical disorders, and TCM use was endorsed as less effective for psychiatric illnesses when compared with physical illnesses. Furthermore, differences in perceived TCM efficacy appeared to be magnified among US-born respondents, with US-born respondents endorsing lower efficacy for psychiatric disorders than foreign-born respondents. These findings suggest that TCM use for psychiatric disorders may decrease with Westernization, but might delay access to psychiatric services among first generation immigrants. PMID- 19282697 TI - Recurrent Escherichia coli bloodstream infections: epidemiology and risk factors. AB - Patients with recurrent episodes of Escherichia coli bloodstream infection (REC BSI) have been described previously only in small studies. We report on the incidence, clinical significance, and predisposing conditions of REC-BSI in a general hospital from 1992 to 2005. All patients with E. coli bloodstream infection (EC-BSI) were retrieved from our database. We defined recurrent episodes as those occurring at least 1 month apart after a clinical response (cases). To study risk factors for REC-BSI, we randomly selected a third of the REC-BSI cases and a similar number of controls (patients with a single EC-BSI). Available E. coli isolates from initial and recurrent episodes were typed using repetitive-extragenic-palindromic-sequences to distinguish between relapse and reinfection. During the study period there were 4287 episodes of EC-BSI in 3970 patients; of these, 251 (6.3%) patients had 568 episodes of recurrence (13.3%). We selected 81 cases and 81 controls for study. The underlying conditions of patients with REC-BSI included immunosuppression (33%), urinary (24%) or biliary obstruction (16%), chronic liver disease (16%), presence of a central venous catheter (8%), and miscellaneous entities (3%). Male sex, presence of hematologic malignancy, inadequate antibiotic treatment, and an extraurinary source of the BSI were independent risk factors for recurrence in the multivariate analysis. Molecular typing performed in 88 infections from 44 patients showed that 47% of REC-BSI were relapses rather than reinfections. Recurrence of E. coli BSI is not an uncommon phenomenon and includes relapses (47%) and reinfections (53%). Recurrence should suggest not only the presence of urinary or biliary obstruction, but also the presence of immunosuppression. PMID- 19282698 TI - Hirschsprung disease and congenital anomalies of the kidney and urinary tract (CAKUT): a novel syndromic association. AB - Congenital anomalies of the kidney and urinary tract (CAKUT) can be associated with Hirschsprung disease (HSCR). Based on the common genetic background of enteric nervous system and kidney development, the reported association of CAKUT and HSCR seems underestimated. Therefore, we designed a prospective study aimed at determining the prevalence of CAKUT in HSCR patients and at identifying RET, glial cell line-derived neurotrophic factor (GDNF), and GDNF family receptor alpha1 (GFRalpha1) mutations or haplotypes associated with this subset of HSCR patients. Eighty-four HSCR patients consecutively admitted to our department between July 2006 and July 2007 underwent interviews, notes review, ultrasound screening (further investigation according to detected anomaly), urinalysis, and DNA extraction for molecular genetics study. Another 27 patients with isolated CAKUT were included as a control group for the molecular genetics study. Twenty one patients (25%) with HSCR had associated CAKUT, with hydronephrosis and hypoplasia being the most frequent diagnoses. Nine of 21 CAKUT were symptomatic. Six additional patients had other non-CAKUT anomalies (for example, stones, Barter syndrome) that were excluded from association and molecular genetics analysis to avoid bias of inclusion criteria. RET mutations were found in 5 patients (4 HSCR, 1 HSCR + CAKUT, 0 CAKUT) and GDNF mutations in 3 (2 HSCR, 1 CAKUT, 0 HSCR + CAKUT). No GFRalpha1 mutations were found. Finally, the HSCR predisposing T haplotype of RET proto-oncogene was found in 64% of HSCR, 50% of HSCR + CAKUT, and in 24% of CAKUT patients. The incidence of CAKUT in HSCR patients is 4- to 6-fold higher than expected. Therefore, a patient with HSCR has a 3- to 18-fold higher risk of developing a CAKUT, particularly hydronephrosis or hypoplasia. If we consider that the proportion of predisposing haplotype in HSCR + CAKUT patients resembles that of other syndromic HSCR, we can conclude that HSCR + CAKUT has to be considered a novel syndromic association. These results need to be confirmed in a larger series. At present, we strongly suggest considering ultrasound screening of the urinary tract in every patient with a diagnosis of HSCR. PMID- 19282699 TI - Factors associated with underlying malignancy in a retrospective cohort of 121 patients with dermatomyositis. AB - Demographic, clinical, and laboratory features that predict underlying malignancy in patients with dermatomyositis (DM) are poorly known. We conducted a retrospective study in all adult patients with a definite (n = 75) or probable (n = 32) diagnosis of DM according to Bohan and Peter criteria or with amyopathic DM (n = 14) who were referred to 2 departments during a 13-year period. The diagnosis of malignancy-associated DM was retained if DM occurred in a context of recently diagnosed malignancy or if a malignancy was diagnosed during the 5 years following the diagnosis of DM. The Kaplan-Meier method was used to assess the cumulative incidence rates of underlying malignancy during the first 5 years of DM. Factors associated with malignancy in patients with DM were identified by Cox proportional hazards models. During the study period, 121 patients fulfilled the inclusion criteria (median age, 52 yr; range, 19-77 yr; women: 70%). For 29 of them, the diagnosis of malignancy-associated DM was retained. The cumulative incidence rate of malignancy was 21 +/- 4% and 28 +/- 5%, 1 year and 5 years after the diagnosis of DM, respectively. The median duration of follow-up of the 92 patients with no malignancy diagnosed was 36 months (range, 1-140 mo). In multivariate analysis, independent factors associated with an underlying malignancy in patients with DM were an age at diagnosis >52 years (hazard ratio [HR], 7.24; 95% confidence interval [CI], 2.35-22.31), a rapid onset of skin and/or muscular symptoms (HR, 3.11; 95% CI, 1.07-9.02), the presence of skin necrosis (HR, 3.84; 95% CI, 1.00-14.85) or periungual erythema (HR, 3.93; 95% CI, 1.16-13.24), and a low baseline level of complement factor C4 (HR, 2.74; 95% CI, 1.11-6.75). Lastly, low baseline lymphocyte count (<1500/mm(3)) was a protective factor of malignancy (HR, 0.33; 95% CI, 0.14-0.80). Taken together, these data may help physicians focus on a group of patients who might benefit from extensive evaluation for malignancy. PMID- 19282700 TI - Renal sarcoidosis: clinical, laboratory, and histologic presentation and outcome in 47 patients. AB - We conducted the current study to investigate the clinical, laboratory, and histologic features at presentation and the outcome of renal sarcoidosis (RS). Exhaustive retrospective data were collected by the French Sarcoidosis Group. Forty-seven adult patients were assessed (30 male/17 female, M/F ratio: 1.76). Median estimated glomerular filtration rate (eGFR) was 20.5 mL/min per 1.73 m(2) (range, 4-93 mL/min per 1.73 m(2)). Moderate proteinuria was found in 31 (66%) patients (median, 0.7 g/24 h; range, 0-2.7 g/24 h), microscopic hematuria in 11 (21.7%) patients, aseptic leukocyturia in 13 (28.7%) patients. Fifteen of 47 (32%) patients had hypercalcemia (>2.75 mmol/L). Eleven of the 22 (50%) patients diagnosed between June and September had hypercalcemia compared with only 4 of the 25 (16%) cases diagnosed during the other months (p < 0.001). Thirty-seven patients presented with noncaseating granulomatous interstitial nephritis (GIN), and 10 with interstitial nephritis without granulomas. Apart from hypercalcemia, the clinical phenotype was also remarkable for the high frequency of fever at presentation. All patients initially received prednisone (median duration, 18 mo), 10 received intravenous pulse methylprednisolone. eGFR increased from 20 +/- 19 to 44 +/- 24.7 mL/min per 1.73 m(2) at 1 month (p < 0.001, n = 38), to 47 +/- 19.9 mL/min per 1.73 m(2) at 1 year (p < 0.001, n = 46), to 49.13 +/- 25 mL/min per 1.73 m(2) at last follow-up (p < 0.001, n = 47). A complete response to therapy at 1 year and at last follow-up was strongly correlated with complete response at 1 month (p < 0.01). Renal function improvement was inversely related to initial histologic fibrosis score. A complete response to therapy at 1 year was strongly correlated with hypercalcemia at presentation (p = 0.003). Relapses were purely renal (n = 3) and purely extrarenal (n = 10) or both (n = 4), often a long time after presentation, with in some cases severe cardiac or central nervous system involvement. We conclude that hypercalcemia and fever at presentation are often associated with RS; RS is most often and permanently responsive to corticosteroid treatment, but some degree of persistent renal failure is highly frequent and its degree of severity in the long run is well predicted from both histologic fibrotic renal score and response obtained at 1 month. PMID- 19282701 TI - Diminished quality of life and physical function in community-dwelling elderly with anemia. AB - The occurrence of anemia in older adults has been associated with adverse outcomes including functional decline, disability, morbidity, and mortality. It is not clear to what extent these outcomes are the result of the anemia or concurrent illness. We performed a cross-sectional, observational study to determine whether lower hemoglobin concentrations in older adults are associated with reduced health-related quality of life, functional status, depression, disability, and physical strength, independent of chronic disease. Three sites participated in this research: an academic geriatric practice, a hospital-based geriatric outpatient unit, and a community-based multispecialty internal medicine group. Health-related quality of life and functional status were measured using the Short Form-36 Health Survey (SF-36) and the Functional Assessment of Chronic Illness Therapy-Anemia (FACIT-An). Disability and depression were assessed using the Instrumental Activities of Daily Living (IADL) and the Geriatric Depression Scale (GDS) questionnaires, respectively. Handgrip strength was used as a physical performance measure. Anemia was defined as hemoglobin <13 g/dL for men or <12 g/dL for women. The mean SF-36 physical health component summary scores were 38.9 (with anemia) and 44.1 (without anemia) (p<0.001). Anemia was associated with greater fatigue (p < 0.001), lower handgrip strength (p = 0.014), increased number of disabilities (p=0.005), and more depressive symptoms (p = 0.002). Multivariate regression analysis, adjusted for demographic and clinical characteristics, demonstrated strong associations for reduced hemoglobin, even within the "normal" range, and poorer health-related quality of life across multiple domains. Thus, anemia was independently associated with clinically significant impairments in multiple domains of health-related quality of life, especially in measures of functional limitation. Mildly low hemoglobin levels, even when above the World Health Organization (WHO) anemia threshold, were associated with significant declines in quality of life among the elderly. PMID- 19282702 TI - Community-acquired bacterial meningitis in elderly patients: experience over 30 years. AB - Clinical characteristics, etiologies, evolution, and prognostic factors of community-acquired bacterial meningitis in elderly patients are not well known. To improve this knowledge, all episodes of community-acquired bacterial meningitis were prospectively recorded and cases occurring in patients >or=65 years old were selected. During the period 1977-2006, 675 episodes in adults (aged >or=18 yr) were recorded, with 185 (27%) in patients aged >or=65 years old; 76 were male and 109 were female, with a mean age of 73 +/- 6 years (range, 65-93 yr). Causative microorganisms were Streptococcus pneumoniae 74, Neisseria meningitidis 49, Listeria monocytogenes 17, other streptococcal 9, Escherichia coli 6, Haemophilus influenzae 4, Klebsiella pneumoniae and Staphylococcus aureus 2 each, Capnocytophaga canimorsus and Enterococcus faecalis 1 each, and unknown in 20. On admission 91% had had fever, 32% were in a coma (Glasgow Coma Scale or=65 yr), who showed a higher frequency of diabetes and malignancy as underlying disease; pneumonia, otitis, and pericranial fistula as predisposing factors; and S. pneumoniae and L. monocytogenes as etiology. There were also differences in clinical presentation, complications, sequelae, and mortality. Factors independently related with mortality were age, pneumonia as a predisposing factor, coma on admission, and heart failure and seizures after therapy. Dexamethasone therapy was a protective factor. In conclusion, bacterial meningitis in elderly patients is associated with greater diagnostic difficulties and neurologic severity and more complications, as well as with increased mortality. Antiseizure prophylaxis might be useful in these patients. PMID- 19282703 TI - Microbiology and outcome of iliopsoas abscess in 124 patients. AB - To describe the microbiology and outcome of iliopsoas abscess (IPA) in a large case series, we analyzed 124 cases of IPA collected from 1990 through 2004 in 11 hospitals in Spain. Twenty-seven (21.8%) patients had primary and 97 (78.2%) had secondary IPA. The main sources of infection were bone (50.5%), gastrointestinal tract (24.7%), and urinary tract (17.5%). A definitive microbial diagnosis was achieved in 93 (75%) cases. Abscess culture was the most frequent procedure leading to microbial diagnosis, followed by blood cultures. Staphylococcus aureus, Escherichia coli, and Bacteroides species were the most frequent microbial causes: S. aureus was the most common organism in patients with primary abscesses (42.9%) and with abscesses of skeletal origin (35.2%), whereas E. coli was the leading organism in those with abscesses of urinary (61.5%) and gastrointestinal (42.1%) tracts. Mycobacterium tuberculosis was found in 15 patients, 4 of them associated with human immunodeficiency virus (HIV) infection. Twenty (21.5%) cases had polymicrobial infections; these were more common among patients with abscesses of gastrointestinal origin. Information on clinical outcome was available for 120 patients; 19 (15.8%) had a relapse and 6 (5%) died due to complications related to the IPA. Patients who died were older and more likely to have bacteremia and E. coli isolated from cultures. In conclusion, secondary IPA is more prevalent than primary IPA. Among those with secondary IPA, most abscesses are secondary to a skeletal source. A bacterial etiology can be identified in most cases. The overall prognosis of patients with this condition is good. PMID- 19282704 TI - The end of postoperative pain--a fast-approaching possibility? And, if so, will we be ready? PMID- 19282705 TI - Designing and implementing a comprehensive learner-centered regional anesthesia curriculum. PMID- 19282706 TI - Continuous femoral nerve block provides superior analgesia compared with continuous intra-articular and wound infusion after anterior cruciate ligament reconstruction. AB - BACKGROUND AND OBJECTIVES: This prospective, randomized, clinical trial compared pain intensity and analgesic drug consumption after anterior cruciate ligament (ACL) reconstruction with patellar tendon under femoral-sciatic nerve block anesthesia in patients who received either a continuous femoral nerve block (CFNB) or continuous local anesthetic wound and intra-articular infusions. METHODS: Fifty patients were randomized to CFNB (n = 25) or an ON-Q device (I Flow Corp, Lake Forest, Calif) (n = 25). All patients received sciatic nerve block (25 mL of ropivacaine 7.5 mg/mL and clonidine 30 microg). The first group received a CFNB (2 mg/mL of ropivacaine at 7 mL/hr), and the second group received a single-shot femoral nerve block (both using 25 mL of ropivacaine 7.5 mg/mL and clonidine 30 microg). At the end of the intervention, an ON-Q device was positioned on the ON-Q patients to continuously infuse the patellar tendon wound and intra-articular cavity with ropivacaine 2 mg/mL at 2 mL/hr for each catheter. Data regarding demographic, hemodynamic, pain scores, adverse effects, and need for supplemental analgesia were registered in a 36-hr follow-up period. RESULTS: The CFNB group reported lower visual analog scale values than the ON-Q group: at rest at 12 hrs (2.4 [SD, 2.2] vs 5.4 [SD, 3.1]; P < 0.001) and on movement at 12 (3.1 [SD, 2.5] vs 6.3 [SD, 2.9]; P < 0.001) and 24 hrs (2.7 [SD, 1.9] vs 4.6 [SD, 2.6]; P = 0.01) after surgery. The number of morphine and ketorolac boluses was lower in the CNFB group (morphine: 3.2 [SD, 2.2] vs 6.2 [SD, 2.5]; P < 0.001; ketorolac: 1.1 [SD, 1.0] vs 2.4 [SD, 0.9]; P < 0.001). CONCLUSION: Continuous femoral nerve block provides better analgesia than the continuous patellar tendon wound and intra-articular infusions after anterior cruciate ligament reconstruction with patellar tendon. PMID- 19282707 TI - A prospective, observational study of the relationship between body mass index and depth of the epidural space during lumbar transforaminal epidural steroid injection. AB - BACKGROUND AND OBJECTIVES: Previous studies have concluded that transforaminal epidural steroid injections (ESIs) are more effective than interlaminar injections in the treatment of radiculopathies due to lumbar intervertebral disk herniation. There are no published studies examining the depth of epidural space using a transforaminal approach. We investigated the relationship between body mass index (BMI) and the depth of the epidural space during lumbar transforaminal ESIs. METHODS: Eighty-six consecutive patients undergoing lumbar transforaminal ESI at the L3-L4, L4-L5, and L5-S1 levels were studied. Using standard protocol, the foraminal epidural space was attained using fluoroscopic guidance. The measured distance from needle tip to skin was recorded (depth to foraminal epidural space). The differences in the needle depth and BMI were analyzed using regression analysis. RESULTS: Needle depth was positively associated with BMI (regression coefficient [RC], 1.13; P < 0.001). The median depths (in centimeters) to the epidural space were 6.3, 7.5, 8.4, 10.0, 10.4, and 12.2 for underweight, normal, preobese, obese I, obese II, and obese III classifications, respectively. Sex (RC, 1.3; P = 0.02) and race (RC, 0.8; P = 0.04) were also significantly associated with needle depth; however, neither factor remained significant when BMI was accounted as a covariate in the regression model. Age, intervertebral level treated, and oblique angle had no predictive value on foraminal depth (P > 0.2). CONCLUSION: There is a positive association between BMI and transforaminal epidural depth, but not with age, sex, race, oblique angle, or intervertebral level. PMID- 19282708 TI - A comparison between EMLA cream application versus lidocaine infiltration for postoperative analgesia after inguinal herniotomy in children. AB - BACKGROUND: EMLA cream (eutectic mixture of local anesthetics) has been shown to penetrate intact skin and provide analgesia of superficial layers. There are no studies on the effects of topical application of EMLA cream for postoperative pain relief after inguinal hernia repair. OBJECTIVE: : This randomized, double blind, placebo-controlled study compared the efficacy of topical application of 5% EMLA cream before surgery, with wound infiltration with 1% lidocaine for postoperative analgesia in children. METHODS: Ninety children, aged 4 to 12 years, undergoing elective inguinal hernia repair under general anesthesia were enrolled in the study. Patients were randomly assigned to receive either placebo cream (group1), 5% EMLA cream (group 2), or placebo cream followed by 0.5 mL/kg 1% lidocaine (group 3) in the wound after induction of anesthesia. The anesthetic technique and monitoring were standardized, and postoperative pain was assessed using a 10-point objective pain scale. Fentanyl was used as rescue analgesic in immediate postoperative period, and acetaminophen was administered for postoperative pain in surgical ward. RESULTS: The number of patients requiring fentanyl in the immediate postoperative period was significantly less in the study groups compared with the placebo group. Sixty-seven percent of patients in the placebo group required more than 1 dose of acetaminophen in the first 6 hrs compared with 23% (EMLA group) and 20% (lidocaine group). Four patients (two in the lidocaine group, one in the EMLA group, and one in the control group) developed subcutaneous infection at the site of incision 10 to 15 days postoperatively. CONCLUSION: Topical application of EMLA (5%) provides postoperative analgesia comparable to infiltration with 1% lidocaine after inguinal hernia repair in children. PMID- 19282709 TI - A web-based cross-sectional epidemiological survey of complex regional pain syndrome. AB - BACKGROUND AND OBJECTIVES: Complex regional pain syndrome (CRPS) is a poorly understood pain disorder with little information on the natural course of the disease. Changes in its diagnostic criteria have simplified the identification of this syndrome, but convincing epidemiological data regarding this disorder are still lacking. Here, we collected epidemiological and other relevant information regarding CRPS via a Web-based survey to develop a better understanding of the epidemiology, symptoms, progression, therapy, and associated psychosocial factors related to CRPS. METHODS: A survey of 75 questions was hosted on the Web site of the Reflex Sympathetic Dystrophy Syndrome Association of America for 5 months. One thousand three hundred fifty-nine subjects responded, and 888 of these satisfied the inclusion criteria for CRPS and were accepted for data analysis. RESULTS: Complex regional pain syndrome affected mostly white women in the 25- to 55-year-old age group. It was often precipitated by trauma (surgical or nonsurgical) and commonly involved the lower ( approximately 56%) and upper ( approximately 38%) extremities. Pain was usually accompanied by edema, vasomotor, sudomotor, motor, and trophic changes. The syndrome commonly progressed and spread to involve other body areas. Affected patients failed multiple pharmacological and nonpharmacological interventions. The syndrome frequently interfered with job ( approximately 62% disability rate), sleep ( approximately 96%), mobility ( approximately 86%), and self-care ( approximately 57%). Remissions and relapses were both common. CONCLUSIONS: Complex regional pain syndrome is a severe disabling pain disorder that results in physical as well as emotional and financial consequences to patients. The disease complexity requires coordination of multidisciplinary care that can be achieved by educational efforts directed to general practitioners. PMID- 19282710 TI - Hydrodynamics of the spinal epidural space in pigs: effects of death and exsanguinations. AB - BACKGROUND AND OBJECTIVES: We have investigated how the vascular components of the spine determine the resistance and capacitance of the spinal epidural space and determined the magnitude of the longitudinal pressure gradient in the space during fluid infusion. METHODS: Pigs were studied during isoflurane anesthesia. Tuohy needles were inserted into midthoracic spine at adjacent interspaces, one to measure pressure in the epidural space and one for fluid infusion. A third Tuohy needle was inserted in the lumbar epidural space. Fluid was infused at a constant flow (0.9-3.1 mL/min) until epidural space pressure reached a steady plateau, then the pump was shut off, and pressure returned to baseline over 5 to 6 mins. Resistance was calculated as the ratio of plateau pressure and flow. Initial (first 20-30 secs) and late (1-5 mins) capacitance was calculated. Measurements were made with the animal alive, 15 mins after its life was terminated by an intravenous injection of KCl, and then after exsanguination. RESULTS: During fluid infusion, the pressure gradient between the lumbar and thoracic epidural space was small, on the order of 1 to 2 mm Hg. Death reduced resistance but not capacitance, whereas exsanguination reduced late capacitance but not resistance. Neither maneuver affected initial capacitance. CONCLUSIONS: There is a small longitudinal pressure gradient within the epidural space during fluid infusion. Hence, the major source of resistance occurs where fluid leaves the epidural space. Death reduced resistance, perhaps by depressurizing spinal arteries in the intervertebral foramina, but did not affect capacitance. Blood in epidural veins is a major determinant of late epidural capacitance. PMID- 19282711 TI - Efficacy of bicarbonate in decreasing pain on intradermal injection of local anesthetics: a meta-analysis. AB - OBJECTIVE: Intradermal injection of local anesthetic often results in pain on injection due in part to the acidic pH of commercially prepared solutions, which are optimized to prolong shelf life. Although there are other possible explanations (eg, noxious properties of local anesthetics, pressure effect of infiltration), the etiology is most likely multifactorial. Although addition of bicarbonate to local anesthetics may decrease pain on intradermal injection, the extent of this analgesic effect is uncertain. We performed a meta-analysis of available trials investigating pain during intradermal injection of buffered local anesthetic preparations. METHODS: We searched the National Library of Medicine's PubMed database for all relevant articles published on the topic through November 2006. Inclusion criteria included double-blind, randomized controlled trials and use of a visual analog scale to measure pain on infiltration of local anesthetic buffered with sodium bicarbonate compared with that of unbuffered local anesthetic. Meta-analysis was performed using the Review Manager 4.2.7 (The Cochrane Collaboration, 2004). A random-effects model was used. RESULTS: Our search resulted in 86 abstracts, of which 12 articles met all inclusion criteria. Overall, there were 609 observations for buffered local anesthetic and 615 for unbuffered local anesthetic. Use of buffered local anesthetic resulted in a statistically lower weighted mean difference in visual analog scale of -1.17 (95% confidence interval, -1.68 to -0.67) compared with unbuffered local anesthetic. CONCLUSIONS: Our systematic review suggests that the use of buffered local anesthetics seems to be associated with a statistical decrease in pain of infiltration when compared with unbuffered local anesthetic. PMID- 19282712 TI - Excretion of ropivacaine in breast milk during patient-controlled epidural analgesia after cesarean delivery. AB - BACKGROUND: Few studies have been published concerning the excretion of bupivacaine and lidocaine into the breast milk and none concerning ropivacaine. AIM: The aim of this study was to determine the levels of ropivacaine in plasma and breast milk after combined spinal-epidural anesthesia for cesarean delivery and postoperative patient-controlled epidural analgesia (PCEA), as well as possible adverse effects from these levels on the neonate. METHODS: Twenty-five parturients admitted for cesarean delivery under combined spinal-epidural anesthesia participated in the study. The PCEA regimen was prepared as 0.15% ropivacaine and fentanyl 2 microg/mL (basal rate, 6 mL/h; demand dose, 4 mL/20 min). Blood samples were collected before anesthesia and from the umbilical cord immediately after birth, followed by blood and breast milk samples 18 and 24 hrs after initiation of PCEA. The newborns were clinically appraised with the Apgar score at delivery and Neurological and Adaptive Capacity Score 24 hrs later. Local anesthetic side effects were recorded. Ropivacaine levels were determined by high-performance liquid chromatography with a UV detector. RESULTS: Ropivacaine excretion into the breast milk produced concentrations that significantly correlated with those in the plasma 18 and 24 hrs after administration, achieving milk-plasma ratios (mean +/- SD) of 0.25 +/- 0.08 and 0.23 +/- 0.07, respectively. Most newborns had maximal Apgar and Neurological and Adaptive Capacity Scores. No adverse effects from ropivacaine were noted in mothers or neonates. CONCLUSIONS: The milk-plasma concentration ratio of ropivacaine was found to be lower than that reported for other local anesthetics It seems that PCEA with ropivacaine/fentanyl after cesarean delivery is not associated with excessive milk-plasma concentrations of ropivacaine. PMID- 19282713 TI - Ketamine as an adjuvant in lidocaine intravenous regional anesthesia: a randomized, double-blind, systemic control trial. AB - BACKGROUND AND OBJECTIVES: Ketamine delays and minimizes intraoperative tourniquet pain when added to lidocaine-based intravenous regional anesthesia (IVRA). It is unclear if adding ketamine to the IVRA injectate is more efficacious compared with systemic administration. This study compares intraoperative tourniquet pain, postoperative analgesia, and side effects of systemic versus IVRA ketamine during outpatient hand surgery. METHODS: We conducted a randomized, double-blind, systemic control study of 40 patients undergoing hand surgery using lidocaine IVRA. In group IVRA, 0.1 mg/kg ketamine in 1 mL of normal saline was added to the IVRA lidocaine, and 1 mL of normal saline was administered via a peripheral IV line. In group systemic, 1 mL of normal saline was added to the IVRA syringe, and 0.1 mg/kg ketamine in 1 mL of normal saline was administered via a peripheral intravenous line. Ten minutes after proximal tourniquet inflation, the distal tourniquet was inflated, and the proximal tourniquet deflated. Tourniquet pain was measured every 10 mins. Need for intraoperative opioids was recorded. Recovery room pain scores, analgesic needs, and sedation scores were compared. Patients were contacted 24 hrs after surgery and reported their analgesic consumption, satisfaction scores, and the occurrence of any unpleasant psychologic effects. RESULTS: Groups IVRA and systemic were comparable in demographic and surgical parameters. There were no differences between groups in intraoperative tourniquet pain scores, intraoperative fentanyl requirements, recovery room pain or sedation scores, postsurgical analgesic needs, or patient satisfaction scores. CONCLUSIONS: In comparison to systemic administration, there is no selective benefit to adding ketamine to the IVRA injectate. PMID- 19282715 TI - Ultrasound-guided supraclavicular block: outcome of 510 consecutive cases. AB - INTRODUCTION: Supraclavicular brachial plexus block provides consistently effective anesthesia to the upper extremity. However, traditional nerve localization techniques may be associated with a high risk of pneumothorax. In the present study, we report block success and clinical outcome data from 510 consecutive patients who received an ultrasound-guided supraclavicular block for upper extremity surgery. METHODS: After institutional review board approval, the outcome of 510 consecutive patients who received an ultrasound-guided supraclavicular block for upper extremity surgery was reviewed. Real-time ultrasound guidance was used with a high-frequency linear probe. The neurovascular structures were imaged on short axis, and the needle was inserted using an in-plane technique with either a medial-to-lateral or lateral-to-medial orientation. RESULTS: Five hundred ten ultrasound-guided supraclavicular blocks were performed (50 inpatients, 460 outpatients) by 47 different operators at different levels of training over a 24-month period. Successful surgical anesthesia was achieved in 94.6% of patients after a single attempt; 2.8% required local anesthetic supplementation of a single peripheral nerve territory; and 2.6% received an unplanned general anesthetic. No cases of clinically symptomatic pneumothorax developed. Complications included symptomatic hemidiaphragmatic paresis (1%), Horner syndrome (1%), unintended vascular punctures (0.4%), and transient sensory deficits (0.4%). CONCLUSIONS: Ultrasound guided supraclavicular block is associated with a high rate of successful surgical anesthesia and a low rate of complications and thus may be a safe alternative for both inpatients and outpatients. Severe underlying respiratory disease and coagulopathy should remain a contraindication for this brachial plexus approach. PMID- 19282716 TI - Ultrasound-guided transsartorial perifemoral artery approach for saphenous nerve block. PMID- 19282718 TI - The conjoint tendon of the latissimus dorsi and teres major: an important landmark for ultrasound-guided axillary block. PMID- 19282714 TI - Upper extremity regional anesthesia: essentials of our current understanding, 2008. AB - Brachial plexus blockade is the cornerstone of the peripheral nerve regional anesthesia practice of most anesthesiologists. As part of the American Society of Regional Anesthesia and Pain Medicine's commitment to providing intensive evidence-based education related to regional anesthesia and analgesia, this article is a complete update of our 2002 comprehensive review of upper extremity anesthesia. The text of the review focuses on (1) pertinent anatomy, (2) approaches to the brachial plexus and techniques that optimize block quality, (4) local anesthetic and adjuvant pharmacology, (5) complications, (6) perioperative issues, and (6) challenges for future research. PMID- 19282719 TI - Use a rifle, not a shotgun. PMID- 19282720 TI - Ultrasound-guided interscalene block should be compared with the accepted standard for the neurostimulation technique. PMID- 19282721 TI - The beneficial role of patient-controlled epidural analgesia in a patient with fournier gangrene. PMID- 19282722 TI - Ultrasound guidance for sciatic nerve block at the popliteal fossa should be compared with the best motor response and the lowest current clinically used in neurostimulation technique. PMID- 19282723 TI - Early detection of intravascular injection during ultrasound-guided axillary brachial plexus block. PMID- 19282724 TI - Transversus abdominis plane block for neuropathic pain. PMID- 19282725 TI - Task-specific frequencies of neck motion measured in healthy young adults over a five-day period. AB - STUDY DESIGN: Observational cohort design. OBJECTIVE: To provide initial estimates of the frequencies and magnitudes of neck motion during daily activities in healthy subjects. SUMMARY OF BACKGROUND DATA: Previous studies have measured the maximum excursions during recreated activities of daily living (ADLs) in laboratory settings, but there is a lack of information available on frequencies and excursions of neck motion with ADLs in nonartificial settings. METHODS: Ten healthy young adults were fitted with a portable motion measurement device that recorded movement about each primary axis. Participants were instructed to wear the unit continuously over a 5-day period and record their daily activities with corresponding times. After the collection period, subjects' activity logs were analyzed and data were partitioned into 5 categories, which provided the most primary representation of ADLs: athletics, work, travel, sleep, and miscellaneous. Each category was further divided into increasingly specific activities (e.g., running and walking). Frequency of motions within 5 degrees increments was determined and an hourly rate was calculated for each activity. Median motion about each axis for each activity was also determined. RESULTS: The total number of movements per hour for all axes, regardless of amplitude, was highest during athletic activity and lowest during sleeping. The majority of movements (92% of athletic activity, 90% of work) required less than 25 degrees of lateral bending, while greater range of movement requirements were observed for flexion-extension and axial rotation. Less than 6% of movements exceeded 50 degrees. The median range of motion along all axes was highest for athletic activity and lowest for sleeping. CONCLUSION: The results of this study provide preliminary observations of the frequency and magnitude of neck motion during normal ADLs for the specified population. These findings can assist physicians and physical therapists in determining the extent of disability and identifying activities that will likely be problematic for patients with limited cervical motion. PMID- 19282726 TI - Different ways to balance the spine: subtle changes in sagittal spinal curves affect regional muscle activity. AB - STUDY DESIGN: Exploratory study of regional muscle activity in different postures. OBJECTIVE: To detail the relationship between spinal curves and regional muscle activity. SUMMARY OF BACKGROUND DATA: Sagittal balanced spinal posture (C7 above S1 in the sagittal plane) is a goal for spinal surgery and conservative ergonomics. Three combinations of thoracolumbar and lumbar spinal curves can be considered sagittal balanced postures: (i) flat-at both regions, (ii) long lordosis-lordotic at both regions, and (iii) short lordosis-thoracic kyphosis and lumbar lordosis. This study compares regional muscle activity between these 3 sagittal balanced postures in sitting, as well as a slump posture. METHODS: Fine-wire electromyography (EMG) electrodes were inserted into the lumbar multifidus (deep and superficial), iliocostalis (lateral and medial), longissimus thoracis, and transversus abdominis in 14 healthy male volunteers. Fine-wire or surface EMG electrodes were also used to record activity of the obliquus internus, obliquus externus, and rectus abdominis muscles. Root mean square EMG amplitude in the flat, long lordosis, short lordosis, and slump sitting postures were normalized to maximal voluntary contraction, and also to the peak activity across the sitting postures. Muscle activity was compared between postures with a linear mixed model analysis. RESULTS: Of the extensor muscles, it was most notable that activity of the deep and superficial fibers of lumbar multifidus increased incrementally in the 3 sagittal balanced postures; flat, long lordosis, and short lordosis (P < 0.05). Of the abdominal muscles, obliquus internus was more active in short lordosis than the other postures (P < 0.05). Comparing the sagittal balanced postures, the flat posture showed the least muscle activity (similar to the slump posture at most muscles examined). CONCLUSION: Discrete combinations of muscle activity supported the 3 different sagittal balanced postures in sitting, providing new detail for surgeons, researchers, and therapists to distinguish between different sagittal balanced postures. PMID- 19282727 TI - Accuracy of dynamic computed tomography to calculate rotation occurring at lumbar spinal motion segments. AB - STUDY DESIGN: Reliability study comparing computed tomography (CT) to biomechanics. OBJECTIVE: To measure the accuracy and precision of such measurements in comparison with a standard method. SUMMARY OF BACKGROUND DATA: Rotations of lumbar spinal motion segments can be measured with dynamic CT imaging. This may be a useful tool to measure intersegmental motion. Validation of its use is lacking. METHODS: Human cadaveric lumbar spines were fixed in a rigid rotation device and rotated, whereas rotation at each level was measured with extensiometers. Rotation at each level was calculated as a percent of total rotation. The spines were placed in a CT scanner and imaged after rotation of the spine in each direction. The percent of total rotation that occurred at each level was calculated with a software program. Accuracy of the CT method was calculated as the average difference between methods. Precision was measured as the standard deviation of the CT measurement. Biomechanical testing and CT were repeated after the posterior anulus fibrosus at L3-L4 was incised with a scalpel. The power of the CT method to detect a change in rotation was tested by calculating the difference between the pre- and postsurgery rotation at L3-L4 and testing it for significance with a Student t test of paired samples. RESULTS: Differences between CT and biomechanical measurements averaged 0.2%. Precision was 6.0% (Table 1). Postanular injury, percent rotation at the L3-L4 level increased, whereas it decreased at the other 4 levels. The change at L3-L4 was statistically significant (P = 0.047). CONCLUSION: Dynamic CT measures vertebral rotations sufficiently accurately to study the effect of a radial tear on axial rotation can. PMID- 19282728 TI - Validation of the Turkish version of the Quebec back pain disability scale for patients with low back pain. AB - STUDY DESIGN: A reliability and validity study of a translated, culturally adapted questionnaire. OBJECTIVE: The aims of the present study were to translate the Quebec Back Pain Disability Scale (QDS) into Turkish, to perform its cross cultural adaptation for Turkish patients with LBP, and to investigate its validity and test-retest reliability. SUMMARY OF BACKGROUND DATA: As a widely used scale in the evaluation of patients with low back pain (LBP), the QDS awaits formal translation and validation into Turkish to achieve an equivalent questionnaire and to allow comparability of data. METHODS: The translation and cross-cultural adaptation of the original questionnaire were performed in accordance with published guidelines. Translation and retranslation of the English version of the QDS was performed blindly and independently by 4 different individuals, and adapted by a team. Hundred patients with LBP were included in our study. The physical examinations were evaluated and the Schober test was assessed for a mobility measurement of the spine. The patients were asked to complete a questionnaire booklet containing the Turkish versions of the modified Oswestry Disability Index (ODI) and QDS, and Visual Analog Scale (VAS) measure of pain. All assessments were repeated 24 hours later for all of the patients. Reliability was evaluated using internal consistency and the intraclass correlation coefficient (ICC). Concurrent validity was measured by comparing the Turkish version of the QDS results to VAS and the Schober test scores. Also, for construct validity, the results of the scale were compared with the Turkish version of modified ODI. RESULTS: The QDS showed excellent test-retest reliability as evidenced by the high ICC for 2 test occasions (ICC = 0.9221, P < 0.000). Also, internal consistency was found to be adequate at both assessments with Cronbach's alpha (0.9405 and 0.9537 at day 0 and 1, respectively). There was a positive correlation between QDS and VAS both for day 0 (r = 0.368; P < 0.000) and for day 1 (r = 0.441; P < 0.000). There was no correlation determined in the comparison of the QDS sum scores with Schober testing for day 0; however, significant negative correlations in these parameters were observed for day 1 (r = -0.249 P = 0.014). Also significantly positive correlations were deter- mined between the Turkish version of the QDS and the Turkish version of the modified ODI for both day 0 and day 1 (r = 0.666, P < 0.000, r = 0.681; P < 0.000, respectively). CONCLUSION: The results of our study show that QDS as a functional status questionnaire has been translated into Turkish without losing the psychometric properties of the original version. The Turkish version of the QDS has good comprehensibility, internal consistency, and validity and is an adequate and useful instrument for the evaluation of disability in patients with LBP. PMID- 19282729 TI - Three rod technique facilitates hemivertebra wedge excision in young children through a posterior only approach. AB - STUDY DESIGN: Retrospective case series. OBJECTIVE: To study the efficacy of a 3 rod technique used in hemivertebra excision via a posterior only approach in young children to avoid implant failure. SUMMARY OF BACKGROUND DATA: Previous reports of posterior only hemivertebra excisions in young children have been associated with an unacceptable amount of implant failures and high revision rates. METHODS: We reviewed our clinical experience in young children who underwent excision of a hemivertebra from a posterior only approach. All children had the wedge resection closed using a rod attached to laminar hooks and had the excision site stabilized with an additional 2 rods attached to pedicle screws. RESULTS: In our series of 10 patients at an average age of 4 years and 3 months, all patients obtained fusion and there were no implant related complications or revisions needed. The average preoperative Cobb measurement of 44 degrees was improved to 8 degrees at most recent follow-up. CONCLUSION: The use of a 3-rod technique during hemivertebra excision in young children helped to avoid any implant failures in our series. We conclude that this technique should be used in young children whose pedicles may not be able to withstand the compressive forces needed to close down the wedge resection site. PMID- 19282730 TI - Selective arterial embolization with n-butyl cyanoacrylate in the treatment of aneursymal bone cyst of the thoracic vertebra: a case report. AB - STUDY DESIGN: A case report of aneurysmal bone cyst of the mobile spine. OBJECTIVE: To present a pediatric case of aneurysmal bone cyst (ABC) of the mobile spine treated successfully by selective arterial embolization (SAE) with n butyl cyanoacrylate (NBCA). SUMMARY OF BACKGROUND DATA: The endovascular treatment is an important treatment option for the ABC of the mobile spine. However, there are few reported case treated by SAE with NBCA. METHODS: A 12-year old healthy boy was introduced with a complaint of back pain. Computed tomography showed an osteolytic expanding mass in the pedicle, lamina, and spinous process on the level of T10. Magnetic resonance imaging revealed a multicystic mass with fluid-fluid levels that extended into the spinal canal and compressed the spinal cord from the dorsal side. Digital subtraction angiography showed an abnormal blood supply from the posterior spinal branch of the left T9 artery. An abnormal mass was diagnosed as an ABC based on these neuroradiologic examinations. RESULT: The patient underwent SAE with NBCA (Histoacryl, B. Braun, Melsungen, Germany) diluted with Lipiodol (Laboratorie Guerbet, France) in a rate of 1:4. Computed tomography performed 1 month after treatment revealed partial ossification of the bone cortex. MR imaging showed shrinkage of the cyst and decompression of the spinal cord. Back pain of the patient disappeared and the shrunken cyst was stable without recurrence during the 3-year follow-up period. CONCLUSION: SAE in a pediatric case with ABC of the mobile spine was effective in improving of clinical symptom and shrinking of the cyst. PMID- 19282731 TI - The vanishment of an intramedullary high-signal intensity lesion at the craniocervical junction after surgical treatment: a case report of the presyrinx state. AB - STUDY DESIGN: A case report of the presyrinx state at the craniocervical junction with vanishment of an intramedullary high-signal lesion followed by decompression and fixation. OBJECTIVE: To report the reversible change of an intramedullary high-signal lesion on T2-weighted MRI as a presyrinx state. SUMMARY OF BACKGROUND DATA: The pathology of a T2-weighted high-signal intensity area in the spinal cord has not yet been described in detail. The case presented here showed the vanishment of this lesion after the surgical procedure, which implies that some high-signal intensity lesions might be reversible as a presyrinx state. METHODS: A 75-year-old man presented with severe cervical myelopathy. Neurologic findings and observations on various images indicated compression myelopathy due to both a pseudotumor at the craniocervical junction and spondylosis at C3-C4 disc level due to Klippel-Feil syndrome. The most obvious finding was a vast high-signal intensity lesion at the craniocervical junction, which was speculated to be a syringomyelia before surgery. RESULTS: The MRI at 1 month after surgical treatment (occipito-spinal fusion with the decompression by enlargement of foramen magnum) revealed complete vanishment of the vast high intensity lesion at the craniocervical junction, which remained undetected at the 2 years and 8 months follow-up and corresponded with improvement in the clinical symptoms of myelopathy. CONCLUSION: The vanishment of these signal changes on MRI after surgery was interpreted as a presyrinx state, demonstrating this reversible pathology in the spinal cord. PMID- 19282733 TI - The effect of calmodulin antagonists on experimental scoliosis: a pinealectomized chicken model. AB - STUDY DESIGN: Randomized controlled. OBJECTIVE: To evaluate the effects of Tamoxifen (TMX) and trifluoperozine (TFP) on pinealectomized chicken scoliosis. SUMMARY OF BACKGROUND DATA: Pinealectomized chicken develops scoliosis probably due to the lack of melatonin. In addition to other functions, melatonin also acts as a calmodulin antagonist. We postulate that loss of this antagonistic effect may be the cause of scoliosis in this model. TMX and TFP are known calmodulin antagonists, which may alter the incidence and severity of scoliosis. METHODS: Seventy-two newly hatched chicken that underwent surgical pinealectomy within 72 hours of hatching were divided into 3 groups of 24 animals in each as group I (control), group II (TMX), and group III (TFP). TMX and TFP were given to groups II and III, respectively, for 10 weeks with the dose of 0.1 mg/kg/d, whereas the control group received no medication. AP scoliosis radiographs were obtained at seventh and 10th week to evaluate coronal spinal alignment. RESULTS: Three chickens in group I, 2 chickens in group II, and 1 chicken in group III died in the first postoperative week. Scoliosis incidences and magnitudes were similar among groups at seventh and 10th week. TMX and TFP groups showed decreases of incidence of upper cervical, lower cervical, lower cervical-thoracic-lumbar curves at 10th week compared with seventh week. TMX group showed a decline in thoracic region mean Cobb angle, whereas control group showed an increase (P = 0.048). TMX group showed a more prominent decline in cervicothoracic region mean Cobb angle compared with control group (P = 0.009). CONCLUSION: The incidence and magnitude of scoliosis in pinealectomized chicken may be decreased by the administration of TMX, presumably because of this drugs' calmodulin antagonism. Further studies on higher animals and dosage and timing are required. PMID- 19282734 TI - Limaprost alfadex improves myelopathy symptoms in patients with cervical spinal canal stenosis. AB - STUDY DESIGN: Myelopathy symptoms were prospectively studied in patients with cervical spinal canal stenosis (CSCS), using objective grading systems and stabilometry, to examine the effect of administration of prostaglandin E1 derivative limaprost alfadex (limaprost). OBJECTIVE: Myelopathy scores/grades and stabilometry parameters were evaluated before, and 1 and 3 months after starting the limaprost treatment. SUMMARY AND BACKGROUND DATA: Limaprost is a potent vasodilator and antiplatelet agent and has been used to treat the symptoms of lumbar spinal canal stenosis. The action presumably involves increased blood flow in the compressed cauda equina. Limaprost can also increase blood flow in the compressed spinal cord, but effects on myelopathy symptoms in patients with CSCS have not been established. METHODS: This study examined 21 patients with mild spondylotic CSCS based on neurologic findings and compression of the cervical spinal cord on magnetic resonance imaging. Japanese Orthopedic Association score, grip and release test, and finger escape sign were measured, and stabilometry was performed by independent examiners, before, and 1 and 3 months after starting the oral limaprost treatment. RESULTS: Most patients experienced amelioration of the symptoms at 1 month after starting the treatment. Mean Japanese Orthopedic Association score and grip and release count were significantly improved and finger escape sign grade was higher in some patients. Stabilometry area with eyes closed and Romberg rate were also significantly improved. These improvements were maintained at 3 months. CONCLUSION: The efficacy of oral limaprost administration for patients with CSCS was confirmed by objective scoring and quantitative data. PMID- 19282735 TI - Dynamics of neurological deficit after surgical decompression of symptomatic vertebral metastases. AB - STUDY DESIGN: We conducted a retrospective study to examination the influence of preoperative duration of symptoms on the clinical outcome of patients that underwent surgical decompression because of neurologic deficit in metastatic disease of the spine. OBJECTIVES.: Our aim was to investigate possible correlations between the duration of neurologic deficit before surgery and postoperative outcome with respect to neural recovery in patients with spinal metastases, and second, based on those results, propose criteria for the timing of surgery in these patients. SUMMARY OF BACKGROUND DATA: It has not yet been determined whether the duration of preoperative symptoms has an influence on the postoperative outcome of patients with vertebral metastases. A standardized treatment or protocol defining a strategy of surgical treatment has yet to be designed. METHODS: This study includes 194 patients. The duration of symptoms before surgical treatment and the neurologic status before and after operation were determined and classified according to the Frankel score. RESULTS: Of 401 patients, who underwent surgery due to metastases to the spine, 194 suffered from neurologic deficit. Analyzing the postoperative neurostatus in these patients revealed an improvement in 78 patients (40%), impairment in 13 patients (7%), and in 103 patients it did not change. The relation of duration of neurologic symptoms before surgery, and the outcome after an operation was highly significant (P < 0.001). In patients with less than 3 days of neurologic deficit, the probability of improvement in neurostatus was highly significantly higher (P < 0.001) than in patients with neurologic deficit existing for more than 15 days. CONCLUSION: Patients with neurologic deficit because of spinal bone metastases benefit from early operative intervention. Urgent surgery is indicated in patients with less than 3 days of neurologic deficit. PMID- 19282736 TI - Significantly improved outcomes with a less invasive posterior lumbar interbody fusion incorporating total facetectomy. AB - STUDY DESIGN: Original study. OBJECTIVE: Prospective comparison of clinical outcomes after a standard posterior lumbar interbody fusion (ST-PLIF) and after a limited exposure PLIF incorporating total facetectomy (LI-PLIF). SUMMARY OF BACKGROUND DATA: Most groups have reported significantly improved clinical outcomes after ST-PLIF. To our knowledge, however, a comparison of outcomes between ST-PLIF and the LI-PLIF that we herein describe has not been reported before. METHODS: Patients were included who had suffered chronic low back pain for a minimum of 2 years that was unresponsive to conservative treatment. N = 114 consecutive patients underwent ST-PLIF, whereas n = 209 underwent LI-PLIF. All patients underwent pre- and postoperative evaluations for Oswestry Disability Index (ODI), short-form 36 (SF-36), and visual analogue scores (VAS). The minimum follow-up for either group was 2 years. RESULTS: There was a significant improvement in the ODI (22.5 +/- 1.0, P < 0.001), VAS for back pain (3.8 +/- 0.1, P = 0.003), VAS for leg pain (4.0 +/- 0.2, P = 0.002), and SF-36 for bodily pain (14.7 +/- 0.9, P = 0.012) after ST-PLIF. However, there was a significantly greater improvement in all scores after LI-PLIF: ODI (28.8 +/- 1.4 vs. 22.5 +/- 1.0, P < 0.001), VAS for back pain (5.4 +/- 0.2 vs. 3.8 +/- 0.1, P = 0.001), VAS for leg pain (5.1 +/- 0.2 vs. 4.0 +/- 0.2, P < 0.001), and SF-36 for bodily pain (18.5 +/- 0.8 vs. 14.7 +/- 0.9, P = 0.003). There was a significantly shorter duration of hospital stay after LI-PLIF (2.24 +/- 0.057 days) than after ST-PLIF (4.04 +/- 0.13 days) (P = 0.005). Operative complications occurred in 19.3% of ST PLIF and in 6.7% of LI-PLIF. CONCLUSION: Clinical outcomes were significantly improved after both ST-PLIF and LI-PLIF. However, outcomes were significantly better after LI-PLIF than after ST-PLIF. Significantly shortened hospital stay with LI-PLIF probably reflected the "less invasive" technique per se. Significantly better clinical outcomes with fewer complications after LI-PLIF, however, potentially reflected maneuvers singular to LI-PLIF: (1) preservation of posterior elements, (2) avoidance of far lateral dissection over the transverse processes, (3) bilateral total facetectomy, (4) fewer neurologic complications, and (5) avoidance of iliac crest autograft. LI-PLIF is therefore recommended over ST-PLIF. PMID- 19282737 TI - Classification of scoliosis deformity three-dimensional spinal shape by cluster analysis. AB - STUDY DESIGN: Cluster analysis of existing database of spinal shape of patients attending a scoliosis clinic. OBJECTIVE: To determine whether patients with scoliosis can be classified into distinct groups by 3-dimensional curve shape. SUMMARY OF BACKGROUND DATA: Subjective or semiquantitative methods can be used to classify curve types in scoliosis, with the goal of rationalizing surgical planning. There are very few reports of using objective methods such as cluster analysis to improve this process. METHODS.: One hundred ten patients who underwent radiography of the spine by a stereo technique, at a scoliosis clinic in the period between 1982 and 1990, were studied. Fifty-six were studied longitudinally (average 3.4 clinic visits each), providing 245 total observations. Selected patients had 2 scoliosis curves with apex between T4 and L3, and both Cobb angles >9 degrees by an automated measurement. The 3 dimensional spinal shape was reconstructed from stereoradiographs. Each curve was quantified by its Cobb angle, apex level, apex vertebra rotation, and rotation of the plane of maximum curvature (PMC) (8 variables). Cluster analysis classified each patient at each visit by these variables. RESULTS: When the analysis searched for 4 clusters, the largest cluster (148 of 245 observations) was the pattern having counterclockwise rotation of the PMC of both curves (typically, a right upper scoliosis curve with kyphosis and left lower scoliosis curve with lordosis). The other 3 clusters (48, 34, and 15 observations) were the other permutations of these variables. Substantial overlap of all the other variables between groups was observed. Of the 56 patients seen longitudinally, 25 were consistently grouped at all clinic visits. CONCLUSION: Spinal shape of patients in a clinic population with 2 scoliosis curves form distinct groups according to the 4 permutations of the signs of the rotations of the PMC in 2 curve regions. The pattern can change with repeated observation, often because a slight curvature in the sagittal plane can change because of postural variation and measurement errors. Overlap of the other curve-shape variables between groups suggests that these spinal deformity classifications alone should not determine treatment strategy. PMID- 19282738 TI - Assessment of spinal flexibility in adolescent idiopathic scoliosis: suspension versus side-bending radiography. AB - STUDY DESIGN: Prospective evaluation of a new suspension test to determine curve flexibility in adolescent idiopathic scoliosis (AIS) in comparison with erect side-bending. OBJECTIVE: To verify whether the suspension is a better method than side-bending to estimate curve reducibility and to assess spine flexibility. SUMMARY OF BACKGROUND DATA: Spinal flexibility is a decisive biomechanical parameter for the planning of AIS surgery. Side-bending is often referred as the gold standard, but it has a low reproducibility and there is no agreement amongst surgeons about the most advantageous method to use. Even more, every technique evaluates reducibility instead of flexibility since the forces involved in the change in shape of the spine are not considered. METHODS: Eighteen patients scheduled for AIS surgery were studied. Preoperative radiological evaluation consisted of 4 radiographs: standing posteroanterior, left and right erect side bending, and suspension. The side-bending and the suspension tests were compared on the basis of the apical vertebrae derotation and the scoliosis curve reduction. Frontal and axial flexibility indices, expressed as the ratio between the moment induced by the body weight and the reduction, were calculated from the suspension data. RESULTS: The average scoliosis curve reduction and apical vertebra derotation were 21 degrees (37%) and 3 degrees (12%), respectively for erect side-bending and 26 degrees (39%) and 7 degrees (28%), respectively for suspension. The erect side-bending test generated a larger curve reduction (P = 0.05) when considering the moderate curves only and the suspension test (P = 0.02) when considering the severe curves. The suspension test produced a larger axial derotation (P = 0.007) when considering all the curves. The average traction force during suspension was 306 N (187 N-377 N). The average estimation for the frontal flexibility index was 1.64 degrees/Nm (0.84-2.82) and 0.51 degrees/Nm (0.01-1.39) for the axial flexibility index. CONCLUSION: Results of this study demonstrate the feasibility to really evaluate the spine flexibility with the suspension test. The estimated flexibility values are realistic and similar to those reported in vitro. Suspension should be used in the future for spine flexibility assessment. PMID- 19282739 TI - Reliability analysis for manual radiographic measures of rotatory subluxation or lateral listhesis in adult scoliosis. AB - STUDY DESIGN: Retrospective observational study. OBJECTIVE: To define the inter- and intraobserver reliability of 3 measures of rotatory subluxation (RS) in adult scoliosis (AS). SUMMARY OF BACKGROUND DATA: RS is a hallmark of AS. To accurately track this measure, one must know its reliability. Reliability testing has not been performed. METHODS: PA 36" films of 29 AS patients were collected from one surgeon's practice. Three observers on 2 separate occasions measured all levels with >or=3-mm RS (60 levels, 360 measurements) on the convexity of the involved segment using 3 different techniques-midbody (MB), endplate (EP), and centroid (C). These data were then analyzed to determine the intraclass correlation coefficient (ICC) for inter- and intraobserver reliability. RESULTS: The thoracolumbar/lumbar curve (average 58 degrees ) was the major curve for the majority (62%) of patients. RS at L3/4 was most common (35%). The overall inter- and intraobserver reliability was good-excellent for all methods, but the centroid method consistently had the highest ICC. ICC correlated with observer experience. Moderate-severe arthritic change (present in 55%) and poor image quality (52%) decreased ICC, but it still remained good-excellent for each measure. The reproducibility coefficient for each measure was 4 mm for MB and 2.8 mm for C and EP. CONCLUSION: MB, EP, and C are reliable techniques to measure RS even in elderly arthritic spines, but the methods inherently produce different values for a given level. The centroid method is most reliable and least influenced by experience. The EP method is easy to perform and very reliable. Spine surgeons should pick their preferred method and apply it consistently. Changes >3 mm suggest RS progression. RS may be a useful measure in addition to Cobb angle in AS. Having defined measurement reliability, the role of RS progression in surgical indications and patient outcomes can be evaluated. PMID- 19282740 TI - Value of immediate preprocedure magnetic resonance imaging in patients scheduled to undergo vertebroplasty or kyphoplasty. AB - STUDY DESIGN: Retrospective case series. OBJECTIVE: To determine the incidence of additional vertebral compression fractures diagnosed with repeat MRI immediately before vertebroplasty or kyphoplasty. SUMMARY OF BACKGROUND DATA: Vertebral compression fractures, which occur frequently in the elderly, are more likely in patients with prior vertebral compression fracture. When patients are evaluated for vertebroplasty, it is important to identify all unhealed fractures for effective treatment planning. METHODS: In a retrospective study, we reviewed the records of all patients (n = 194) treated with vertebroplasty or kyphoplasty over a 6-year period at our institution, and identified all patients who had undergone a repeat MRI within 7 days of vertebroplasty or kyphoplasty. These studies were obtained as part of a clinical protocol prescribing a repeat MRI for any patient whose MRI had been obtained more than 3 months before the evaluation, or who had a change in their pain between referral and evaluation. RESULTS: Twenty patients met inclusion criteria for the immediate preprocedure MRI protocol. A total of 14 new fractures in 11 patients were discovered on the immediate preprocedure MRI. Of these 14, 6 had less than 15% loss of height, making them potentially occult on radiographs; 3 fractures developed in 2 patients who had no change in back pain. CONCLUSION: In select candidates for vertebroplasty or kyphoplasty, a repeat preprocedure MRI obtained within 1 week can help ensure that all painful fractures are treated. There is demonstrable value in this protocol for patients with an imaging evaluation over 3 months old or who have had any change in symptoms since the initial imaging workup. PMID- 19282741 TI - Point of view. Efficacy of vertebroplasty and kyphoplasty. PMID- 19282742 TI - Spine and pain clinics serving North Carolina patients with back and neck pain: what do they do, and are they multidisciplinary? AB - STUDY DESIGN: Cross-sectional survey. OBJECTIVE: Our primary objective was to describe spine and pain clinics serving North Carolina residents with respect to organizational characteristics. Our secondary objective was to assess the multidisciplinary nature of the clinics surveyed. SUMMARY OF BACKGROUND DATA: Pain clinics have become common in the United States, and patients with chronic back pain have increasingly been seeking services at these clinics. Little is known about the organizational characteristics of spine and pain clinics. METHODS: We identified and surveyed spine and pain clinics serving North Carolina residents with chronic back and neck pain. Practice managers at 46 clinics completed a 20-minute questionnaire about the characteristics of their clinic, including providers on staff and services offered. Descriptive and exploratory analyses were conducted to summarize the data. Several variables were constructed to assess the multidisciplinary nature of the clinics. RESULTS: The response rate was 75%. There was marked heterogeneity among the clinics surveyed. Fifty-nine percent of practices were free-standing (n = 27) and 61% were physician-owned (n = 28). Twenty-five clinics (54%) had an anesthesiologist. Other common physician providers were physiatrists and surgeons. Less than one third of sites had mental health providers (n = 12; 26%); only 26% employed physical therapists. Seventy six percent of sites offered epidural injections, 74% long-term narcotic prescriptions, and 67% antidepressants. The majority of clinics (30 of 33) prescribing narcotics provided monitoring of therapy using periodic urine toxicology testing. Forty-eight percent of sites (n = 22) offered exercise instruction. Few clinics were multidisciplinary in nature. Only 3 (7%) met the criteria of having a medical physician, registered nurse, physical therapist, and mental health specialist. CONCLUSION: Clinics varied widely in their organizational characteristics, including providers and scope of services available. Few clinics were multidisciplinary in nature. This information should be used to determine how pain clinics can better serve patients and improve outcomes. PMID- 19282743 TI - Reliability and validity of adapted French Canadian version of Scoliosis Research Society Outcomes Questionnaire (SRS-22) in Quebec. AB - STUDY DESIGN: Prospective validation study of a cross-cultural adaptation of the Scoliosis Research Society (SRS) Outcomes Questionnaire. OBJECTIVE: To provide a French Canadian version of the SRS Outcomes Questionnaire and to empirically test its response in healthy adolescents and adolescent idiopathic scoliosis (AIS) patients in Quebec. SUMMARY OF BACKGROUND DATA: The SRS Outcomes Questionnaire is widely used for the assessment of health-related quality of life in AIS patients. METHODS: French translation and back-translation of the SRS-22 (SRS-22-fv) were done by an expert committee. Its reliability was measured using the coefficient of internal consistency, construct validity with a factorial analysis, concurrent validity by using the short form-12 and discriminant validity using ANOVA and multivariate linear regression, on 145 AIS patients, 44 patients with non clinically significant scoliosis (NCSS), and 64 healthy patients. RESULTS: The SRS-22-fv showed a good global internal consistency (AIS: Cronbach alpha = 0.86, NCSS: 0.81, and controls: 0.79) and in all of its domains for AIS patients. The factorial structure was coherent with the original questionnaire (47.4% of explained variance). High correlation coefficients were obtained between SRS-22 fv and short form-12 corresponding domains. Boys had higher scores than girls, scores worsened with age, and with increasing body mass index. Mean Total, Pain, Self-image, and Satisfaction scores, were correlated with Cobb angle. Adjusted regression models showed statistically significant differences between the AIS, NCSS, and control groups in the Total, Pain, and Function scores. CONCLUSION: The SRS-22-fv showed satisfactory reliability, factorial, concurrent, and discriminant validity. This study provides scores in a significant group of healthy adolescents and demonstrates a clear gradient in response between subjects with AIS, NCSS, and controls. PMID- 19282744 TI - Percutaneous aspiration of spinal terminal ventricle cysts using real-time magnetic resonance imaging and navigation. AB - STUDY DESIGN: Clinical case series. OBJECTIVE: To present a new innovative treatment option for cysts of the terminal ventricle. SUMMARY OF BACKGROUND DATA: Invasive surgery including laminectomy and cyst fenestration has been the sole reported method of treatment for this rare pathology. METHODS: Four cases of symptomatic cyst of the terminal ventricle of the spinal cord were treated by percutaneous aspiration using real-time guidance with magnetic resonance imaging (MRI) and optically-tracking navigation. RESULTS: Three patients were treated successfully by this percutaneous method, and for 1 patient the treatment was converted to conventional surgery after a single unsuccessful trial of percutaneous puncture. All patients were relieved of their symptoms without any complication. Follow-up MRI revealed continued shrinkage of the cysts after both surgery and MRI-guided percutaneous fenestration. CONCLUSION: MRI-navigated percutaneous aspiration can be a minimally-invasive treatment option for symptomatic cysts of the terminal ventricle, and therefore, can be indicated before more invasive surgery might be scheduled. PMID- 19282745 TI - Surgical site infection in spinal metastasis: risk factors and countermeasures. AB - STUDY DESIGN: A retrospective review (phase 1) and prospective clinical study (phase 2). OBJECTIVES: To identify independent risk factors for surgical site infection (SSI) and to evaluate the positive effect of prostaglandin E1 (PGE1) to decrease the risk of SSI in patients with spinal metastasis. SUMMARY OF BACKGROUND DATA: Surgery for spinal metastasis is associated with an increased risk of SSI. Although previous reports have evaluated risk factors of SSI for spinal metastasis, most of the studies lack multivariate analysis. A recent study demonstrated the utility of PGE1 in decreasing wound complications in patients with prior irradiation. The role of PGE1 in surgery for spinal metastasis has not been previously evaluated. METHODS: One hundred ten patients with spinal metastasis were retrospectively reviewed (phase 1). Risk factors for SSI were analyzed using logistic regression. Phase 2 was a prospective clinical trial investigating the utility of PGE1 at reducing the rate of SSI. Ninety-four patients with spinal metastasis were treated at our institute. The infection rate and risk factors identified in phase 1 and 2 were compared. RESULTS: The rate of SSI during phase 1 was 7.1%. Independent risk factors identified by multivariate logistic regression were diabetes, and preoperative irradiation. The rate of SSI for patients who had irradiation before surgery was 32%, whereas the rate for patients without irradiation was 1.1%. This difference was statistically significant. The rate of SSI in phase 2 was 3.1%. In phase 2 patients who received preoperative irradiation, the rate of SSI was 4.5%. The difference between phase 1 and phase 2 was statistically significant. CONCLUSION: This study identified diabetes and preoperative irradiation to be independent risk factors for SSI in patients with spinal metastasis. PGE1 administration was found to significantly decrease the incidence of SSI in patients with spinal metastasis who underwent preoperative irradiation. PMID- 19282746 TI - Clinical design functions: round table discussions on the bioengineering of liquid ventilators. AB - During the 6th International Symposium on Perfluorocarbon Application and Liquid Ventilation, a round table discussion on bioengineering was held in which different experts shared their opinions and experiences about the use of a total liquid ventilator design for clinical applications. To structure the discussion, all experts were invited to contribute their knowledge within the context of three matrixes related to the liquid ventilators: 1) function and technology, 2) ventilation modes, and 3) risk analyses. The outcome of this international conference recommends continued development of a total liquid ventilator toward clinical applications. PMID- 19282747 TI - Biocompatibility study of hemoglobin vesicles, cellular-type artificial oxygen carriers, with human umbilical cord hematopoietic stem/progenitor cells using an in vitro expansion system. AB - Hemoglobin vesicles (HbVs), liposomal oxygen carriers containing human hemoglobin, are candidates for development of a clinically useful transfusion alternative. Our previous in vivo animal studies of massive HbV dosages demonstrated the lack of any suppressive effect on hematopoiesis. Before starting clinical trials, we aimed to examine the details of the biocompatibility of HbVs with human hematopoietic stem/progenitor cells. We investigated the effect of HbVs at a concentration of up to 3 vol/vol (%) on expansion of human umbilical cord (CB) hematopoietic stem/progenitor cells, using a coculture system of human TERT-transfected bone marrow stromal cells and CD34+ cells in vitro. The exposure of CB CD34+ cells to HbVs up to 14 days suppressed the expansion of total cells and the CD34+ cells themselves, whereas the empty liposomes, that did not contain Hb, had modestly inhibitory effects on the expansion of these cells. As a result, the number of colonies obtained from the expanded CD34+ cells was inhibited by the exposure to HbVs. In contrast, exposure to HbVs for 3 days had no effect on the expansion of CD34+ cells and only slightly decreased the number of total cells. Our in vitro experimental condition does not fully recreate the physiological condition, and the effect of the direct contact of HbV would be magnified because of the absence of shielding by the vasculature and the lack of the reticuloendothelial system and blood stream. However, the present data raise some concern regarding hematopoiesis, and one has to pay attention to this in future human clinical trials. PMID- 19282748 TI - Lack of correlation of glucose levels in filtered blood plasma to density and conductivity measurements. AB - The purpose of this research project was to determine whether the glucose level of a blood plasma sample from a diabetic patient could be predicted by measuring the density and conductivity of ultrafiltrate of plasma created by a 30,000 m.w. cutoff membrane. Conductivity of the plasma filtrate measures electrolyte concentration and should correct density measurements for changes in electrolytes and water concentration. In vitro studies were performed measuring conductivity and density of solutions of varying glucose and sodium chloride concentrations. Plasma from seven hospitalized patients with diabetes was filtered across a 30,000 m.w. cutoff membrane. The filtrate density and conductivity were measured and correlated to glucose levels. In vitro studies confirmed the ability to predict glucose from density and conductivity measurements, in varying concentrations of glucose and saline. In plasma filtrate, the conductivity and density measurements of ultrafiltrate allowed estimation of glucose in some patients with diabetes but not others. The correlation coefficient for the combined patient data was 0.45 which was significant but only explained 20% of the variability in the glucose levels. Individually, the correlation was significant in only two of the seven patients with correlation coefficients of 0.79 and 0.88. The reasons for lack of correlation are not clear, and cannot be explained by generation of idiogenic osmoles, effects of alcohol dehydrogenase, water intake, etc. This combination of physical methods for glucose measurement is not a feasible approach to measuring glucose in plasma filtrate. PMID- 19282749 TI - Clinical application of pulsatile perfusion during cardiopulmonary bypass in pediatric heart surgery. AB - The benefits of pulsatile over nonpulsatile perfusion has been widely debated in pediatric cardiac operations with cardiopulmonary bypass (CPB). To evaluate the role of pulsatile perfusion in pediatric complicated patients with congenital heart disease undergoing open heart surgery, we performed pulsatile CPB and compared several effects with nonpulsatile perfusion. Pediatric patients (n = 24) diagnosed as typical tetralogy of Fallot (TOF) were randomly divided into two groups: pulsatile perfusion (PP) group and nonpulsatile perfusion (NP) group. Pulsatile perfusion patients used modified roller pump PP during cross-clamping period in CPB, although NP cases used roller pump continuous flow perfusion during CPB. We monitored hemodynamic status and inflammatory media in blood samples over time in all patients. Effective PP can be monitored in PP patients and pulse pressure (DeltaP) was significantly higher in PP group than NP group (p < 0.01). Inflammatory media peaked at the time CPB was weaned off. In PP patients, IL-8 and TNF-alpha were lower after cross-clamp off and intensive care unit period than in NP cases. Free plasma hemoglobin concentration in PP group at preclamp off and CPB weaned off were higher than that of NP group (p < 0.05). Pulsatile perfusion can be successfully applied in pediatric perfusion. Pulsatile perfusion had the role of reducing concentration of inflammatory media in pediatric patients. PMID- 19282750 TI - Single center experience with a low volume priming cardiopulmonary bypass circuit for preventing blood transfusion in infants and small children. AB - This retrospective study analyzed the current practice of blood transfusion-free open-heart surgery in 536 children weighing 5-20 kg undergoing surgery between 2004 and 2007. A miniaturized cardiopulmonary bypass (CPB) circuit was used (priming volume; 300 ml for the flow rate <1,500 ml/min; 550 ml for the flow rate of 1500-2300 ml/min). Modified ultrafiltration was routinely performed. Criteria for blood transfusion during CPB included a hematocrit of <20% and/or mixed venous oxygen saturation of <65%. Transfusion during CPB was avoided in 264 (49.3%) of the 536 patients (5-10 kg group, 29.0%; 11-15 kg group, 67.4%; 16-20 kg group, 80.8%). There was no neurological complication related to hemodilution. Multiple logistic regression analysis revealed that body weight, preoperative hematocrit, priming volume of CPB circuit, CPB time, and lowest hematocrit during CPB predict requirement of blood transfusion (p < 0.01). Transfusion rate was lowest in the atrial septal defect group (5.6%) and highest in tetralogy of Fallot group (78.7%), being associated with complexity of diagnosis and procedure required. Blood transfusion-free open-heart surgery may be achieved in the half of the patients weighing 5-20 kg, and further miniaturization of CPB circuit and refinement of perfusion strategy might reduce transfusion rate in patients <10 kg and/or with complex congenital heart disease. PMID- 19282751 TI - First experience with a paracorporeal artificial lung in humans. AB - Lung transplantation is the only treatment option for patients suffering form end stage respiratory failure. To date, no mechanical device is available to support patients on the waiting list up to months. Here, we summarize our experience with our first two patients, who were supported with a paracorporeal artificial lung (PAL) placed in parallel to the pulmonary circulation with connection to the pulmonary artery and to the left atrium. A low resistance membrane oxygenator (iLA, Novalung, Hirrlingen, Germany) was attached in both patients. Our first patient suffering from a pulmonary veno-occlusive disease was supported for 18 days until he died due to severe sepsis. Our second patient with a primary pulmonary hypertension of unknown origin was supported 62 days followed by successful lung transplantation. In conclusion, the experience obtained with these first two patients under PAL encourages further studies and introduction of this promising concept into clinical practice. PMID- 19282752 TI - Near-infrared spectroscopy as a guide for an intermittent cerebral perfusion strategy during neonatal circulatory arrest. AB - In view of the existing controversy concerning the best perfusion technique during deep hypothermic circulatory arrest (DHCA) for neonatal heart operations, we examined intraoperative rSO2 to help define an optimal interval for an intermittent antegrade cerebral perfusion (IACP) strategy. Records of patients undergoing stage 1 palliation (S1P) and repair of total anomalous pulmonary venous return (rTAPVR) from 1996 to 2004 were reviewed. A total of 16 patients were identified (11 S1P, 5 rTAPVR) with complete data and long periods of DHCA. A decline in rSO2 of either 20% or below a value of 50 was considered significant. The rSO2 for all patients was evaluated after 5, 10, 15, and 20 minutes of DHCA for significant cerebral desaturation. The average rSO2 at the start of DHCA ranged from 45 to 89 for S1P and 35-86 for rTAPVR. Significant cerebral desaturation was observed in 25%-31% of patients after 5 minutes; 42%-44% of patients after 10 minutes; 58%-69% after 15 minutes; and 75%-83% after 20 minutes. Each neonate has a unique baseline cerebral saturation. Also, the response to DHCA varies among subjects as the rate of decrease of rSO2 was not uniform. Universally applying the same interval after which to perfuse the brain permits significant cerebral desaturation in a large percentage of patients. Cerebral oximetry may provide a guide for developing an individualized cerebral perfusion strategy. PMID- 19282753 TI - Middle-molecule clearance in CRRT: in vitro convection, diffusion and dialyzer area. AB - Several studies have attempted to compare different doses and modes of therapy in continuous renal replacement therapies in critically ill patients. It is commonly asserted in the literature that convective therapies can achieve higher clearance of middle molecules than achieved by dialysis alone. However, regardless of the actual prescription, most therapies will actually contain a mixture of both diffusive and convective clearance. Molecular transport in purely convective prescriptions may be hindered by clotting and protein interactions with the dialyser. We measured middle molecule clearance using a tracer molecule, Ficoll, in citrated bovine blood. Using a 2 x 2 factorial design, we examined the impact of prescription [postdilution continuous venovenous hemofiltration (CVVH) vs. continuous venovenous hemodialysis (CVVHD)] and membrane area (0.4 m2 vs. 2.0 m2) on blood-side and dialysate-side middle-molecule clearance. In large dialysers, convective and diffusive prescriptions resulted in nearly identical middle molecule clearance from 10 to 100 kDa molecular weight. In the smaller dialyser, middle molecule clearance was higher when a diffusive therapy (CVVHD) was prescribed versus a convective therapy (postdilution CVVH). We hypothesized that high ultrafiltration rates in the smaller dialyzer resulted in a concentration polarization at the membrane that formed a prefilter, limiting middle-molecule clearance. This effect has implications for design and analysis of clinical trials of continuous renal replacement therapy (CRRT). PMID- 19282754 TI - Dysmorphic findings in two cases of abeta/hypobetalipoproteinemia. PMID- 19282755 TI - Trisomy 9p and Prader-Willi syndromes in an infant resulting from a de-novo unbalanced t(9;15) translocation. AB - Trisomy 9p is a well-described dysmorphic syndrome. The physical features include hypertelorism, down-slanting palpebral fissures, deep-set eyes, down-turned corners of the mouth, and mild skeletal anomalies including hypoplastic terminal phalanges. We report an infant born with some of the typical features of trisomy 9p syndrome, as well as additional features that include extreme joint hyperlaxity with subluxation of the knees and elbows, arachnodactyly, and total anomalous pulmonary venous return. The karyotype revealed an unbalanced chromosome complement. Specifically, a derivative chromosome from a de-novo unbalanced translocation of chromosomes 9 and 15 resulted in partial trisomy of 9pter to 9q13 and deletion of the long arm of chromosome 15 proximal to band q13. Fluorescence in-situ hybridization studies and methylation analysis by Southern blotting revealed deletion of the SNRPN locus on the paternally derived chromosome 15, consistent with Prader-Willi syndrome. This infant represents the first reported case of trisomy 9p syndrome with total anomalous pulmonary venous return and hypoplasia of the amygdala and hippocampus, with the additional finding of Prader-Willi syndrome resulting from a derivative chromosome arising from an unbalanced de-novo t(9;15) translocation. PMID- 19282756 TI - Multiple pterygium syndrome with horseshoe kidney and polydactyly: a further case. PMID- 19282757 TI - Arginase activity in carcinoma of the gallbladder: a pilot study. AB - Carcinoma of the gallbladder is the third most common cancer of the gastrointestinal tract. Recent studies have shown increased arginase activity in various malignancies. The main aim of this study was to evaluate whether arginase activity increases in carcinoma of the gallbladder. The arginase activity was evaluated in serum and gallbladder tissue in 22 patients with histologically proven carcinoma of the gallbladder and 20 patients with cholecystitis using spectrophotometry and western blot assay. The Student's t-test, analysis of variance, and Student-Newman-Keuls test were used for comparison of data and for statistical significance. The mean tissue arginase and serum arginase activity (118.64+/-17.45 and 15.91+/-1.91, respectively) in cases of carcinoma of the gallbladder were significantly higher in comparison with cholecystitis (86.37+/ 4.45 and 12.73+/-0.72, respectively). Subgroup analysis showed stage III gallbladder carcinoma had the maximum tissue arginase activity (142.00+/-21.68 U/g of tissue) followed by stage II (124.15+/-19.88) and stage I (108.46+/-6.73). This significant rise in mean tissue arginase and serum arginase activity in patients with gallbladder cancer probably supports an association between arginase activity and the malignancy. PMID- 19282758 TI - Thymidine kinase activity in serum of renal cell carcinoma patients is a useful prognostic marker. AB - It is known that the concentration and activity of the DNA precursor enzyme thymidine kinase 1 (TK1) in serum is significantly elevated in patients with malignancies, as compared with levels in patients with benign tumours and those in healthy individuals. For the first time, the use of serum TK1 as a prognostic marker for patients with renal cell carcinoma (RCC) was examined. Serum TK1 protein (STK1p) concentration and serum TK1 activity (STK1a) were determined by a dot blot chemoluminescence assay and a radio enzyme assay, respectively. There was no correlation between STK1p and STK1a in the same sera from 27 RCC patients. Only one STK1p value as compared with 15 STK1a values was clearly above the cut off values (2 pmol/l and 6 U/l, respectively) for healthy individuals. STK1a values did not correlate with the level of TK1 expression in tumour sections from the RCC patients, estimated by immunohistochemistry staining. However, there was a significant correlation between STK1a levels and the grade, stage and size of the RCC tumours. The discrepancy between the STK1p and the STK1a results is likely to be because of reduced ability of the TK1 antibody to recognize the STK1 in sera from RCC patients. We conclude that the activity of STK1 is a useful tool for evaluating the prognosis of patients with RCC. PMID- 19282759 TI - Outcome of brain trauma patients who have a Glasgow Coma Scale score of 3 and bilateral fixed and dilated pupils in the field. AB - OBJECTIVE: To investigate the outcome of brain trauma patients who had a Glasgow Coma Scale score (GCS) of 3 and bilateral fixed and dilated pupils (BFDP) in the field. METHODS: Between January 2001 and December 2005, 13 European centres enrolled patients with severe brain trauma. Data sets of all patients who had a GCS of 3 as well as BFDP were analysed. Patients were classified according to the Glasgow Outcome Scale, 12 months after trauma as 'good' (Glasgow Outcome Scale of 5 or 4) or 'poor' functional recovery; relevant data for these two groups were compared. Variables that showed differences in univariate analyses (chi and Wilcoxon-Mann-Whitney tests) were then used as covariates in logistic regression models. A P value of less than 0.05 was considered significant. RESULTS: Ninety two (7.8%) of 1172 patients had a GCS of 3 and BFDP; eight had 'good', 84 had 'poor' recovery. We found no significant differences in sex (79% male), age (median 32 years), and trauma mechanisms. Trauma was significantly less severe, probability of survival significantly higher (0.48 vs. 0.23) in the 'good' group. Only one of 39 patients who had closed basal cisterns on the first computed tomography scan, and none of the patients with midline shift greater than 15 mm had good outcomes. Logistic regression revealed that age, trauma severity, and status of basal cisterns on the first computed tomography scan were the factors determining outcomes. CONCLUSION: Patients with a GCS of 3 and BFDP in the field should be resuscitated aggressively, especially if the trauma seems to be not too severe. PMID- 19282760 TI - Adult epiglottitis: 6 years experience in a university teaching hospital in Hong Kong. AB - Adult epiglottitis is a relatively rare but potentially lethal disease. It seems to be increasing in incidence in other countries. The objective of this study was to examine the common presentations and clinical course of adult epiglottitis in Hong Kong. Retrospective review of 80 consecutive cases of adult epiglottitis admitted between 2000 and 2005. A rising trend in the incidence of adult epiglottitis was observed. The mean age (SD) was 49 years (15.7) and the male-to female ratio was 2.5:1. Disease presentation was nonspecific and sore throat was the most common symptom. All patients were admitted to hospital; 47 (59%) patients were admitted to the intensive care unit (ICU) and 31 (39%) had provision of an artificial airway. Mean (SD) length of stay in ICU and hospital was 1.9 (2.2) and 6.3 (2.9) days, respectively. Six patients (7.5%) were complicated by epiglottic abscess formation. No deaths occurred in this series. Patients who required airway intervention had a higher ICU admission rate and a higher bacterial identification rate. Flexible laryngoscopy was more sensitive than lateral neck radiography in diagnosing epiglottitis (100 vs. 81.4%). The laryngoscopic finding of a narrowed airway was strongly related to the requirement for airway intervention (odds ratio=23.7, 95% confidence interval=4.2 132.6, P<0.001). A high index of suspicion is required for making the diagnosis of adult epiglottitis. Proper airway assessment and close monitoring are of the utmost importance in patient management. Flexible laryngoscopy is the key investigation of choice in the emergency department. PMID- 19282761 TI - Cost awareness among Swedish physicians working at the emergency department. AB - BACKGROUND: In Sweden the public health system is subject of increasing budget problems. Medical training in Sweden does not yet focus on the cost aspects in patients diagnoses and treatment. STUDY OBJECTIVES: This study was carried out to survey the general knowledge among physicians working in the emergency department about the cost of the usual diagnostic tests used in ruling out pulmonary embolism. METHODS: Knowledge about costs among Swedish physicians was evaluated in the context of the emergency department of a university hospital. Thirty physicians answered an anonymous questionnaire about the costs for several items used in the diagnosis of pulmonary embolism. Answers within +/-25% of the real costs were regarded as correct. Evaluation took into consideration the physicians' specialty and experience as emergency physician, house officer or consultant in internal medicine. Statistical analysis was carried out using the Student's t-test, Kruskal-Wallis test and Mann-Whitney U test where applicable. Probability levels of less than 0.05 were accepted as significant. RESULTS: Mean deviation to the real cost was 52% with a correct estimation of an average 28%. No significant difference could be detected in this study among emergency department physicians, house officers and consultants in internal medicine regarding the cost awareness towards the items considered. CONCLUSION: Generally cost awareness was considered low among all participating doctors. The level of experience seemed not to be related to better knowledge about prices. Younger doctors generally tended to underestimate the costs of investigation. Further education will be necessary to raise the level of cost awareness among hospital doctors. PMID- 19282762 TI - Reliability and validity of two four-level emergency triage systems. AB - OBJECTIVE: To measure and compare the reliability and predictive validity of a four-level triage system (I-4L) and the new four-level model triage emergency method (TEM). METHODS: This observational study was conducted in an urban hospital. Ten nurses were randomly selected to assign a triage level to 189 paper scenarios, using either the I-4L model (5 nurses) or the TEM model (5 nurses). We used weighted kappa statistics to measure the interrater and intrarater reliability of each triage tool and assessed the validity of each models based on the accuracy in predicting admission. RESULTS: Interrater reliability was kappa=0.73 [95% CI (confidence interval): 0.59-0.87] and kappa=0.79 (95% CI: 0.65 0.93) with I-4L and TEM, respectively. Intrarater reliability was kappa=0.82 (95% CI: 0.67-0.96) and kappa=0.78 (95% CI: 0.62-0.93), respectively. The accuracy of triage rating for admission prediction was similarly good with I-4L and TEM, namely, 79% (95% CI: 74-85) and 77% (95% CI: 74-85). The proportion of patients admitted per triage level was similar with the two models. CONCLUSION: The interrater and intrarater reliability for rating triage acuity and for accuracy in patient admission prediction was good with both models. Performance with the new model was similar to that of I-4L despite the nurses' short experience. The new TEM model has the advantage of predicting utilization of emergency department resources. PMID- 19282763 TI - Efficacy and safety of microwave ablation for primary and secondary liver malignancies: a systematic review. AB - This article reviews the therapeutic efficacy and complications of microwave ablation (MWA) in the treatment of primary and secondary liver malignancies. A PubMed search using keywords 'microwave', 'liver', 'malignancy', 'cancer' and 'tumour' was performed to identify articles related to MWA of liver malignancies published in English from 1975 to February 2008. MWA is an effective treatment options for both primary and secondary liver malignancies with survivals comparable with those of liver resections. Local recurrences can be managed with further ablation. Small tumour size, well-differentiated tumour and a reduced number of lesions are factors associated with good prognosis. Temporary occlusion of the portal venous and hepatic arterial flow may increase the size of ablation but the safety aspect requires further validation. MWA is a minimally invasive technique that has broadened the therapeutic option for patients with conventionally unresectable liver tumours with promising survival data. Future advances in the applicator design and treatment monitoring may further improve its efficacy and widen the indications. PMID- 19282764 TI - Patients with nonalcoholic fatty liver disease display increased serum resistin levels and decreased adiponectin levels. AB - AIM: Resistin and adiponectin are recently discovered protein hormones, which are produced and secreted by adipocytes. Nonalcoholic fatty liver disease (NAFLD) is a metabolic syndrome, which is associated with obesity. The aim of this study was to evaluate the changes of serum adiponectin and resistin in patients with NAFLD and to determine the relationship between serum adipokine levels and clinicopathologic parameters of NAFLD. METHODS: Forty-three patients with NAFLD and 43 controls were enrolled in this pair-matched study. Body weight, height, body mass index, abdominal wall fat thickness, waist circumference, hip circumference, and the percentage of body fat were measured. Additionally, serum lipid, glucose, alanine aminotransferase, aspartate aminotransferase, adiponectin, and resistin were determined in all individuals. Serum adiponectin and resistin levels were determined using ELISA kits. RESULTS: Serum adiponectin levels were significantly lower in patients with NAFLD compared to the control group (control: 2.01+/-1.10 mg/l vs. NAFLD: 1.38+/-0.65 mg/l, P<0.01). Serum resistin levels were significantly elevated in patients with NAFLD compared to the control group (control: 4.70+/-3.30 ng/ml vs. NAFLD: 9.20+/-7.20 ng/ml, P<0.05). Serum adiponectin concentration was negatively correlated with the waist circumference (rho=-0.425), body mass index (rho=-0.329), percentage of body fat (rho=-0.256), abdominal wall fat thickness (rho=-0.226), and fasting blood glucose concentration (rho=-0.242), but was positively correlated with HDL (rho=0.226). Serum resistin concentration was positively correlated with waist circumference (rho=0.237). No correlation was found between resistin levels and blood pressure, fasting blood glucose concentration, triglyceride, total cholesterol, and HDL. CONCLUSION: NAFLD patients had lower adiponectin levels and higher resistin levels. A positive correlation was found between resistin and waist circumference, whereas a negative correlation was found between adiponectin and waist circumference, body mass index, percentage of body fat, abdominal wall fat thickness, and fasting blood glucose concentration. These data suggested that hypoadiponectinemia and hyperresistinemia might be involved in the development of NAFLD. PMID- 19282765 TI - Impact of gastroesophageal reflux disease on the quality of life of Spanish patients: the relevance of the biometric factors and the severity of symptoms. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) has a negative impact on health-related quality of life (HRQoL). AIM: (i) To evaluate HRQoL in the general Spanish population with GERD. (ii) To analyze the influence of biometric factors and symptoms profile on HRQoL. MATERIALS AND METHODS: A multicenter and population-based evaluation was performed. Surveys were sent out to the general population and 335 responders (16%) fulfilled symptomatic criteria of GERD. They filled out two HRQoL questionnaires [Short Form-36 (SF-36) and Quality of Life in Reflux and Dyspepsia]. Biometric factors and symptoms profile were collected. The SF-36 physical and mental component summary scores were calculated. RESULTS: Two hundred and fifty-two individuals (75.2%) participated. SF-36 and Quality of Life in Reflux and Dyspepsia (QoLRAD) punctuations were lower compared with the general population. Female sex, severity of symptoms, and nocturnal symptoms were associated with a greater deterioration of HRQoL. Obese participants had a negative impact on the physical component summary score, but mental component summary score was better than in participants with normal weight. Adjusted by age, punctuations of smokers, exsmokers, and nonsmokers were similar. No differences in HRQoL with regard to alcohol consumption were observed. CONCLUSION: In participants who fulfill symptomatic criteria of GERD, the HRQoL is very deteriorated. The factors that worsen the QoL are being female, increase in BMI, and nocturnal symptoms. PMID- 19282766 TI - Overlap syndrome triggered by acute viral hepatitis A. AB - We report the case of a 24-year-old woman with an autoimmune hepatitis/primary biliary cirrhosis overlap syndrome triggered by an acute hepatitis A. A number of viruses have been proposed as potential triggers of autoimmune hepatitis in patients with genetic predisposition. To date, approximately 10 cases of type 1 autoimmune hepatitis following hepatitis A virus infection have been published in the medical literature. To our knowledge, this is the first case of overlap syndrome triggered by an acute hepatitis A. PMID- 19282767 TI - Intrahepatic cholangiocarcinoma arising in multiple bile duct hamartomas: report of two cases and review of the literature. AB - Multiple bile duct hamartomas (BDHs)/von Meyenburg complexes, are tumor-like lesions of the liver. Malignant transformation in BDHs has been previously reported in very rare instances, and the most common tumor arising in this clinical setting is cholangiocarcinoma. Herein, we report on clinicopathological findings in two cases of cholangiocarcinoma occurring in liver with multiple BDHs. Histopathologically, multiple BDHs showed morphologic transition from clearly benign to dysplasia or carcinoma in situ, then to invasive carcinoma sequence of the biliary epithelium. The neoplastic epithelium showed positivity for cytokeratin 19, CA 19-9, and epithelial membrane antigen. Staining for Hep Par 1, alpha-fetoprotein, cytokeratin 20, and alpha1-antitrypsin was negative. All sections from the non-neoplastic liver in each specimen showed multiple BDHs. Any other clinically detectable primary tumor was not found. These two neoplasms were interpreted as a cholangiocarcinoma arising in BDHs. This suggested BDHs might be a risk factor of development of cholangiocarcinoma. PMID- 19282768 TI - Percutaneous transhepatic choledochoscopic lithotomy as a rescue therapy for removal of bile duct stones in Billroth II gastrectomy patients who are difficult to perform ERCP. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography (ERCP) is more difficult and dangerous in patients with Billroth II (B II) gastrectomy than those with normal anatomy. OBJECTIVES: To evaluate the clinical efficacy of percutaneous transhepatic choledochoscopic lithotomy (PTCSL) for removing common bile duct stones in B II gastrectomy patients who are difficult to perform ERCP. METHODS: This study was designed as prospectively uncontrolled in a large tertiary referral center. In 20 patients, mostly men, with bile duct stones and an earlier B II gastrectomy, PTCSL was tried because of failed ERCP and high risk. The PTCSL was performed using electrohydraulic lithotripsy or papillary balloon dilation. Successful stone removal and complications were measured. RESULTS: Stone removal was achieved in all 20 patients. The mean number of procedures and session time were 4.5 and 45 min, respectively. Minor PTCSL related complications, such as fever, hemobilia, hyperamylasemia, and wound pain, occurred in five patients (25%). There were no major procedure-related complications, including perforation or mortality. CONCLUSION: The PTCSL procedure is an effective and safe rescue therapy for common bile duct stones in B II gastrectomy patients with failed ERCP and high risk. PMID- 19282769 TI - Long-term follow-up of patients with iron deficiency anaemia after a negative gastrointestinal evaluation. AB - OBJECTIVES: Our aim was to determine whether patients who have had a negative gastrointestinal evaluation (i.e. oesophagogastroduodenoscopy and a colonic examination) for iron deficiency anaemia are subsequently found to have recurrent anaemia or significant pathology. METHODS: From a prospectively entered endoscopy database, we identified a cohort of patients who had negative upper and lower gastrointestinal (GI) investigations for iron deficiency anaemia. We carried out a retrospective chart review of these patients to determine their outcome after a GI evaluation. In particular, we wished to determine the proportion of patients who had recurrent anaemia, became transfusion dependent or were found to have significant pathology. RESULTS: Sixty-nine patients, with an average age of 65.8 years (range 29-87), were followed up for a median of 5 years and 10 months (range 7-109). In 57 patients (83%), the anaemia resolved after the initial treatment period. Fifteen patients (22%) died during the follow-up period, two from a GI cancer and 13 from non-GI-related causes. Six patients (9%) developed persistent anaemia severe enough to require recurrent blood or iron transfusions. Seventeen patients (25%) had a transient recurrent anaemia and four (6%) were diagnosed with GI malignancies during the follow-up. CONCLUSION: For the majority of patients with the iron deficiency anaemia and a negative GI evaluation the outcome is favourable, although a proportion (6%) may subsequently be found to have significant GI pathology. We believe that this number could be minimized by the use of colonoscopy rather than barium enema. In addition, small bowel investigations should not be limited to those who are transfusion dependent, as is currently recommended. PMID- 19282770 TI - Diffuse enteritis after colectomy for ulcerative colitis: two case reports and review of the literature. AB - Ulcerative colitis (UC) is an inflammatory disease of the colon. Involvement of the small bowel is limited to backwash ileitis or pouch-related conditions. Here, we report two men with UC who presented with small bowel inflammation and even perforation, within 1 month after subtotal colectomy. Endoscopy showed diffuse enteritis. Histology showed marked apoptosis of epithelial cells in both cases. One patient responded to steroids and the other to a calcineurin inhibitor. Both patients had no evidence of Crohn's disease in the small intestine before this event. Several more cases of small intestinal lesions in patients with well established UC have been reported. The majority typically presented shortly after colectomy and responded well to steroids. The pathogenesis of this enteritis is unknown, but seems to be distinct from Crohn's disease and may be associated with UC and colectomy. PMID- 19282771 TI - Side-viewing endoscope for colonic self-expandable metal stenting in patients with malignant colonic obstruction. AB - Malignant colon obstruction is a medical emergency and endoscopic decompression, achieved by self-expandable metal stent placement, has been broadly used for both palliation in patients with advanced, nonresectable carcinoma and as a bridge to surgery in those patients with resectable disease. We propose stent insertion assisted by a side-viewing endoscope, in sharply angulated and remarkably stenotic lesions located at the distal sigmoid. Endoscopists should consider, at least for lesions situated in the distal colon, the insertion of a side-viewing endoscope for stenting of technically difficult cases of malignant intestinal obstruction. PMID- 19282772 TI - Methylation of helicase-like transcription factor in serum of patients with colorectal cancer is an independent predictor of disease recurrence. AB - BACKGROUND: CpG island hypermethylation is a common epigenetic event in colorectal cancer. The presence of simultaneous methylation of multiple genes is associated with poor prognosis in many types of tumours including colorectal cancer. We have shown earlier that the hypermethylation of the genes HLTF and HPP1/TPEF are independent prognostic serum markers in colorectal cancer identifying patients with increased risk of death. The purpose of this study was to analyse whether these factors also identify patients at risk of disease recurrence after curative surgery. METHODS: Pretherapeutic sera of 106 patients curatively resected for colorectal cancer with known 5-year follow-ups were analysed for the presence of methylation of the genes HLTF and HPP1/TPEF. RESULTS: HLTF serum methylation was associated with an increased risk of disease recurrence by a factor of 2.7 (95% confidence interval: 1.2-6.0; P=0.014). Multivariate analysis showed methylated HLTF serum DNA to be independently associated with poor outcome and a relative risk of disease recurrence of 2.5 (95% confidence interval: 1.1-5.6; P=0.023). CONCLUSION: Here, we show for the first time that a DNA methylation-based surrogate marker can serve as a predictor of disease recurrence in colorectal cancer. PMID- 19282773 TI - AsktheGeneticist: five years of online experience. AB - PURPOSE: To identify the genetic informational needs and assess the level of awareness about clinical genetic services among adults who use the internet. METHODS: We created an online service called AsktheGeneticist (http://www.askthegen.org) to answer questions about medical genetics. Since 2003, we have received 4497 questions from every US state and 84 countries/territories. Genetic counselors draft answers to the questions submitted. The questions and answers are next reviewed by clinical geneticists, then organized by topic and uploaded to the site. A link to an online website user satisfaction survey is e-mailed to the user with a link to their Q&A. RESULTS: Before visiting AsktheGeneticist, 20% (50/247) of survey respondents were unaware that genetic services existed. After visiting our website, 23.5% (58) of survey respondents sought contact with a genetics health care professional, compared with <1% of patients who self-refer to a general genetics clinic (binomial test; P < 0.0001). Website users most often sought information about a known genetic condition in their family and the risk of recurrence. CONCLUSIONS: Our data suggest that the internet can be an effective tool for increasing the awareness of genetic services and identifying genetic informational needs of online adults, as well as for connecting patients with genetic services. PMID- 19282775 TI - The interface of Medicare coverage decision-making and emerging molecular-based laboratory testing. AB - The process of obtaining Medicare coverage for clinical services (both at the national and local levels) can be complex and often leads to considerable confusion among external stakeholders. The entry of molecular diagnostic testing into the clinical arena of laboratory medicine has posed some special challenges, both for those providing the testing, and those paying for such technology. This commentary will seek to clarify Medicare's pursuit of defining medical necessity by describing both the local and national Medicare coverage policy processes. However, it should be understood that the Medicare reimbursement for such esoteric testing is a work-in-progress, without an established step-by-step process for obtaining a positive coverage decision. Yet, this evolving process provides all stakeholders (payers, laboratories, industry, clinicians, etc.) with an opportunity to fully understand the health policy implications of complex molecular diagnostic testing. In addition, brief case study vignettes are incorporated into our discussion, to show how laboratorians, in conjunction with their clinical colleagues, can effectively engage the payer community in developing more medically sound and fiscally responsible coverage policies. PMID- 19282774 TI - Prevalence and nonrandom distribution of exonic mutations in interferon regulatory factor 6 in 307 families with Van der Woude syndrome and 37 families with popliteal pterygium syndrome. AB - PURPOSE: Interferon regulatory factor 6 encodes a member of the IRF family of transcription factors. Mutations in interferon regulatory factor 6 cause Van der Woude and popliteal pterygium syndrome, two related orofacial clefting disorders. Here, we compared and contrasted the frequency and distribution of exonic mutations in interferon regulatory factor 6 between two large geographically distinct collections of families with Van der Woude and between one collection of families with popliteal pterygium syndrome. METHODS: We performed direct sequence analysis of interferon regulatory factor 6 exons on samples from three collections, two with Van der Woude and one with popliteal pterygium syndrome. RESULTS: We identified mutations in interferon regulatory factor 6 exons in 68% of families in both Van der Woude collections and in 97% of families with popliteal pterygium syndrome. In sum, 106 novel disease-causing variants were found. The distribution of mutations in the interferon regulatory factor 6 exons in each collection was not random; exons 3, 4, 7, and 9 accounted for 80%. In the Van der Woude collections, the mutations were evenly divided between protein truncation and missense, whereas most mutations identified in the popliteal pterygium syndrome collection were missense. Further, the missense mutations associated with popliteal pterygium syndrome were localized significantly to exon 4, at residues that are predicted to bind directly to DNA. CONCLUSION: The nonrandom distribution of mutations in the interferon regulatory factor 6 exons suggests a two-tier approach for efficient mutation screens for interferon regulatory factor 6. The type and distribution of mutations are consistent with the hypothesis that Van der Woude is caused by haploinsufficiency of interferon regulatory factor 6. On the other hand, the distribution of popliteal pterygium syndrome-associated mutations suggests a different, though not mutually exclusive, effect on interferon regulatory factor 6 function. PMID- 19282776 TI - Targeted comparative genomic hybridization array for the detection of single- and multiexon gene deletions and duplications. AB - PURPOSE: To develop a high resolution microarray based method to detect single- and multiexons gene deletions and duplications. METHODS: We have developed a high resolution comparative genomic hybridization array to detect single- and multiexon deletions and duplications in a large set of genes on a single microarray, using the NimbleGen 385K array with an exon-centric design. RESULTS: We have successfully developed, validated, and implemented a targeted gene comparative genomic hybridization arrays for detecting single- and multiexon deletions and duplication in autosomal and X-linked disease-associated genes. CONCLUSION: The comparative genomic hybridization arrays can be adopted readily by clinical molecular diagnostic laboratories as a rapid, cost-effective, highly sensitive, and accurate approach for the detection of single- and multiexon deletions or duplications, particularly in cases where direct sequencing fails to identify a mutation. PMID- 19282777 TI - A tough-love pedagogy in rehabilitation: integration of rehabilitation ideology with local cultures. AB - This study problematizes a unique therapeutic relationship in rehabilitation and how the interaction reflects the integration of rehabilitation ideology with local cultures. The data drew from a larger ethnographic study of a rehabilitation unit in Taiwan. Participants included 21 patient-caregiver pairs and their rehabilitation professionals. They participated in in-depth interviews and participant observation. A tough-love pedagogy emerged as a unique therapeutic relationship in the unit. Patients were asked to interpret the stress with therapy as an inevitable, beneficial experience toward recovery. A prevalent supposition that equated poor physical performance with weak morale legitimized the approach. Cultural metaphors used to describe and define rehabilitation transformed the stress that patients experienced with strenuous exercises into a beneficial substance that aids recovery. The transformation of the therapeutic relationship into a pedagogical one helped connect rehabilitation to shared educational experiences. In the unit, the complicit practice of therapists, caregivers, and patients established and perpetuated the practice of a tough-love pedagogy. The congruence between this tough-love approach and traditional Chinese pedagogical principles made the approach legitimate and desired. PMID- 19282778 TI - Risk of myocardial infarction and abacavir therapy: no increased risk across 52 GlaxoSmithKline-sponsored clinical trials in adult subjects. AB - BACKGROUND: Recently, the Data collection of Adverse events of Anti-HIV Drugs Group (D:A:D) described results from their international observational cohort of 33,347 HIV-1-infected individuals, suggesting unexpected increased risk of myocardial infarction (MI) associated with abacavir (ABC) therapy [relative rate 1.9, 95% confidence interval (CI): 1.47 to 2.45; P = 0.0001]. To contribute to the scientific question, we summarized GlaxoSmithKline HIV clinical trial data to determine if a similar signal emerged. METHODS: We compiled data from GlaxoSmithKline-sponsored clinical trials with > or = 24 weeks of combination antiretroviral therapy comprising 14,174 HIV-infected adults who received ABC (n = 9502; 7641 person-years) or not (n = 4672; 4267 person-years). FINDINGS: Baseline demographics and HIV disease characteristics, including lipids and glucose values, were similar. MI rates were comparable among subjects exposed [n = 16 (0.168%; CI: 0.096 to 0.273; 2.09 per 1000 person-years)] or not [n = 11 (0.235%; CI: 0.118 to 0.421; 2.57 per 1000 person-years)] to ABC-containing therapy. Results of 12 trials with randomization to ABC or not were consistent (2.15 per 1000 person-years vs. 4.10 per 1000 person-years). INTERPRETATIONS: In this pooled summary, we observed few MI events overall and no excess risk of MI with ABC therapy. It is unclear why results from this data set seem discrepant to the Data collection of Adverse events of Anti-HIV Drugs data set, particularly, as the non-ABC MI event rate is similar. Further data are needed to evaluate any association between ABC and increased risk of MI. PMID- 19282779 TI - Abacavir and metabolite pharmacokinetics in HIV-1-infected children and adolescents. AB - OBJECTIVES: Abacavir (ABC) oral clearance, adjusted for body size, is approximately 2 times higher for children than adults with a corresponding difference in dose regimens. However, there are limited data available in the adolescent population. The pharmacokinetics (PKs) of ABC and primary metabolites were determined in HIV-1-infected children and adolescents to evaluate age and patient characteristics as a basis for adjusting ABC dose regimens and to assess the influence of metabolite formation on PK parameters. METHODS: Pediatric subjects 9-18 years of age receiving antiretroviral therapy for HIV-1 infection were stratified by Tanner stage and given a single 8 mg/kg dose of ABC oral solution. Blood samples (n = 10) were obtained over 8 hours and measured for ABC, glucuronide, and carboxylate metabolites using high-performance liquid chromatography. PK parameters for children (Tanner stages 1-2; TS1) and adolescents (Tanner stages 3-5; TS2) were compared. RESULTS: Twenty-five subjects were enrolled. ABC mean (range) maximum concentration (Cmax; microg/mL), area under the curve (microg.hr/mL), half-life (hours), and apparent clearance (CL/F; mL/min per kg) for TS1 and TS2 were 3.5 (1.2-5.6) vs 3.4 (1.8-5.9), 8.0 (2.1 18.6) vs 8.9 (3.1-17.2), 1.3 (0.7-2.5) vs 1.4 (0.9-1.9), and 22.1 (7.0-59.2) vs 18.4 (7.7-42.9) and not significantly different. Age, Tanner stage, and sex were not correlated with ABC clearance by univariate analysis. The ratios of metabolites to ABC area under the curve were correlated with ABC clearance as were the ratios of metabolites to ABC concentrations at the 6-hour time point. CONCLUSIONS: ABC oral clearance in HIV-1-infected pediatric patients does not change during puberty, is similar to younger children, and is higher than previously published in adults. Therefore, dosing adolescents as adults should be reexamined. Intersubject PK variability is substantial and is not correlated with body size or age but more likely due to differences in metabolite formation that may be genetic in origin. PMID- 19282780 TI - Antiretroviral therapy adherence, virologic and immunologic outcomes in adolescents compared with adults in southern Africa. AB - OBJECTIVE: To determine adherence to and effectiveness of antiretroviral therapy (ART) in adolescents vs. adults in southern Africa. DESIGN: Observational cohort study. SETTING: Aid for AIDS, a private sector disease management program in southern Africa. SUBJECTS: Adolescents (age 11-19 years; n = 154) and adults (n = 7622) initiating ART between 1999 and 2006 and having a viral load measurement within 1 year after ART initiation. MAIN OUTCOME MEASURES: Primary: virologic suppression (HIV viral load < or = 400 copies/mL), viral rebound, and CD4 T-cell count at 6, 12, 18, and 24 months after ART initiation. Secondary: adherence assessed by pharmacy refills at 6, 12, and 24 months. Multivariate analyses: loglinear regression and Cox proportional hazards. RESULTS: A significantly smaller proportion of adolescents achieved 100% adherence at each time point (adolescents: 20.7% at 6 months, 14.3% at 12 months, and 6.6% at 24 months; adults: 40.5%, 27.9%, and 20.6% at each time point, respectively; P < 0.01). Patients achieving 100% 12-month adherence were significantly more likely to exhibit virologic suppression at 12 months, regardless of age. However, adolescents achieving virologic suppression had significantly shorter time to viral rebound (adjusted hazard ratio 2.03; 95% confidence interval: 1.31 to 3.13; P < 0.003). Adolescents were less likely to experience long-term immunologic recovery despite initial CD4 T-cell counts comparable to adults. CONCLUSIONS: Compared with adults, adolescents in southern Africa are less adherent to ART and have lower rates of virologic suppression and immunologic recovery and a higher rate of virologic rebound after initial suppression. Studies must determine specific barriers to adherence in this population and develop appropriate interventions. PMID- 19282781 TI - Psychological well-being among individuals aging with HIV: the value of social relationships. AB - OBJECTIVE: Utilizing a heterogenous sample of adults diagnosed with HIV infection, the current study sought to explore associations among age, various dimensions of social support, and psychological and functional well-being. METHODS: Cross-sectional data capturing subjective and instrumental support, social interaction, behavioral health service utilization, and psychological well being (ie, positive affect and depressive symptomatology), and physical functioning, were collected from 109 men and women living with HIV. To explore age group differences, participants were stratified by age (<54 vs. 55+ years). RESULTS: Despite endorsing greater medical comorbidity, older adults reported significantly lower depressive symptomatology and greater positive affect and were less likely to report seeing a behavioral health specialist than their younger counterparts. No age group differences emerged for instrumental support or amount of social interaction. However, older adults reported higher subjective support, which in turn was associated with lower depressive symptomatology, greater positive affect, and nonutilization of behavioral health services. CONCLUSIONS: More attention should be paid to the social environment of individuals diagnosed with HIV as the quality of social relationships may be particularly important for successful psychological adaptation to HIV. PMID- 19282783 TI - Estimating HIV prevalence and risk behaviors among high-risk heterosexual men with multiple sex partners: use of respondent-driven sampling. AB - OBJECTIVES: To collect HIV data from high-risk men who have multiple, younger, female sex partners in a periurban township in South Africa. DESIGN: Unlinked anonymous cross-sectional survey using respondent-driven sampling. METHODS: Survey conducted among men aged 18 years or older who reported having had sex with more than 1 female partner in the previous 3 months (one of whom was either 3 or more years younger than the participant or below the age of 24) and lived in the area of recruitment. RESULTS: The median age of the 421 recruited men was 28 years (range: 18-62 years). They reported a median of 6 sexual partners (range: 2 39) during the past 3 months, and 51% (confidence interval: 45.0 to 59.6) reported inconsistent condom use with their casual partners. During the 3 months before the survey, 98% of men reported having concurrent sexual relationships. HIV prevalence was 12.3% (confidence interval: 8.3% to 16.9%). Being older than 24 years and not using a condom during the last sexual intercourse with a 1-time sexual partner were significantly associated with HIV infection. CONCLUSIONS: This group of heterosexual urban men practice high levels of risky sexual behavior and are an important group that require more targeted HIV surveillance and prevention interventions. PMID- 19282782 TI - Response to zidovudine/didanosine-containing combination antiretroviral therapy among HIV-1 subtype C-infected adults in Botswana: two-year outcomes from a randomized clinical trial. AB - BACKGROUND: Numerous national antiretroviral (ARV) treatment initiatives offering protease inhibitor-sparing combination antiretroviral therapy (cART) have recently commenced in southern Africa, the first of which began in Botswana in January 2002. Evaluation of the efficacy and tolerability of various protease inhibitor-sparing cART regimens requires intensive study in the region, as does investigation of the development of drug resistance and the optimal means of sustaining adherence. The "Tshepo" Study is the first large-scale, randomized, clinical trial that addresses these important issues among HIV-1 subtype C infected ARV treatment-naive adults in southern Africa. METHODS: The Tshepo Study is a completed, open-labeled, randomized study that enrolled 650 ARV-naive adults between December 2002 and 2004. The study is a 3 x 2 x 2 factorial design comparing the efficacy and tolerability among factors: (1) 3 combinations of nucleoside reverse transcriptase inhibitors (NRTIs): zidovudine (ZDV) + lamivudine (3TC), ZDV + didanosine (ddI), and stavudine (d4T) + 3TC; (2) 2 different nonnucleoside reverse transcriptase inhibitors (NNRTIs): nevirapine and efavirenz; and (3) 2 different adherence strategies: the current national "standard of care" versus an "intensified adherence strategy" incorporating a "community-based directly observed therapy." Study patients were stratified into 2 balanced CD4 T-cell count groups: less than 201 versus 201-350 cells per cubic millimeter with viral load greater than 55,000 copies per milliliter. Following Data Safety Monitoring Board recommendations in April 2006, ZDV/ddI-containing arms were discontinued due to inferiority in primary end point, namely, virologic failure with resistance. We report both overall data and pooled data from patients receiving ZDV/ddI- versus ZDV/3TC- and d4T/3TC-containing cART through April 1, 2006. RESULTS: Four hundred fifty-one females (69.4%) and 199 males with a median age of 33.3 years were enrolled into the study. The median follow-up as of April 1, 2006, was 104 weeks, and loss to follow-up rate at 2 years was 4.1%. The median baseline CD4 T-cell count was 199 cells per cubic millimeter [interquartile ratio (IQR) 136-252], and the median plasma HIV-1 RNA level was 193,500 copies per milliliter (IQR 69-250, 472-500). The proportion of participants with virologic failure and genotypic resistance mutations was 11% in those receiving ZDV/ddI-based cART versus 2% in those receiving either ZDV/3TC- or d4T/3TC-based cART (P = 0.002). The median CD4 T-cell count increase at 1 year was 137 cells per cubic millimeter (IQR 74-223) and 199 cells per cubic millimeter (IQR 112-322) at 2 years with significantly lower gain in the ZDV/ddI arm. At 1 and 2 years, respectively, 92.0% and 88.8% of patients had an undetectable plasma HIV-1 RNA level (< or = 400 copies/mL). Kaplan-Meier survival estimates at 1 and 2 years were 96.6% and 95.4%. One hundred twenty patients (18.2%) had treatment-modifying toxicities, of which the most common were lipodystrophy, anemia, neutropenia, and Stevens-Johnson syndrome. There was a trend toward difference in time to treatment-modifying toxicity by pooled dual NRTI combination and no difference in death rates. CONCLUSIONS: The preliminary study results show overall excellent efficacy and tolerability of NNRTI-based cART among HIV-1 subtype C-infected adults. ZDV/ddI-containing cART, however, is inferior to the dual NRTIs d4T/3TC or ZDV/3TC when used with an NNRTI for first line cART. PMID- 19282785 TI - Congenital bilateral sternocleidomastoid contracture: a case report. AB - Unilateral sternocleidomastoid muscle contracture causing torticollis and other secondary deformities such as facial scoliosis, plagiocephaly and scoliosis of cervical spine are well known. The aetiology and pathogenesis is still intriguing. Although unilateral contracture of sternocleidomastoid is seen quite often, bilateral sternocleidomastoid contracture is almost unheard of. A review of the English literature revealed no cases of bilateral congenital sternocleidomastoid contracture being reported. We present a case report of a 19 year-old girl with congenital bilateral sternocleidomastoid contracture. PMID- 19282784 TI - HIV-1 transmission cluster with T215D revertant mutation among newly diagnosed patients from the Basque Country, Spain. AB - OBJECTIVE: To determine the introduction of HIV-1 genetic forms and to examine transmission clusters and resistance to antiretroviral inhibitors among newly diagnosed patients from the Basque Country, Spain, during 2004-2007. METHODS: A total of 261 samples, corresponding to 47.5% heterosexuals, 37.9% men who have sex with men (MSM), and 11.1% intravenous drug users were analyzed in protease and reverse transcriptase to examine phylogenetic relationships and drug resistance-associated mutations. RESULTS: Subtype B was detected in 220 (84.3%) samples and non-B subtype variants in 41 (15.7%) samples. Nearly half (47%) of the sequences grouped in transmission clusters. One of these comprised 14 individuals, 12 of them MSM, with the T215D revertan mutation. In largest transmission clusters, the percentage of MSM was higher than heterosexuals (P < 0.001). Resistance mutations were detected in 29 (11.1%) patients: 20 (7.6%) of them to nucleoside reverse transcriptase inhibitor; 6 (2.3%) to nonnucleoside reverse transcriptase inhibitor (NNRTI); and 1 each to protease inhibitors, protease inhibitor plus NNRTI, and nucleoside reverse transcriptase inhibitor plus NNRTI, respectively. CONCLUSIONS: Our findings underscore recommendations for HIV-1 genotyping in newly diagnosed patients not only to provide information on transmitted drug resistance as an issue in public health and as a guide to future therapy but also to document transmission clusters and to increase the necessary preventive measures. PMID- 19282786 TI - Predictors of outcome of distal rectus femoris transfer surgery in ambulatory children with cerebral palsy. AB - The objective of this study was to identify the predictors of outcome of distal rectus femoris transfer in cerebral palsy. Preoperative and postoperative gait data for 81 patients were examined, focusing on knee flexion/extension range. Outcome was 'good' for 46 patients and 'poor' for 35. The poor outcome group had no improvement in knee range because of increased crouch postoperatively. Outcome was unrelated to quadriceps strength, crouch, velocity, or type of cerebral palsy. Gross Motor Function Classification System was predictive of outcome, with poor results in all level IV patients (P< or =0.008). In conclusion, Gross Motor Function Classification System IV patients may not benefit from distal rectus femoris transfer because of increased postoperative crouch. PMID- 19282787 TI - Results of treatment of idiopathic clubfoot in older infants using the Ponseti method: a preliminary report. AB - The Ponseti method has become a well-established technique for the treatment of clubfoot presenting in the neonatal period. A few reports have discussed the result of this method in older age group. The purpose of this study is to present the results and clinical experience of using the Ponseti method in the treatment of idiopathic congenital talipes equinovarus in infants presented between 4 and 13 months of age with a history of failed manipulations. Thirty-two feet in 20 infants (12 males; eight females) with idiopathic congenital clubfeet were treated using the Ponseti method with minor modifications. The average age at presentation was 7 months (range from 4 to 13 months). We used the Pirani scoring system to assess the feet. After an average follow-up of 19 months, the ultimate overall results were satisfactory in 31 feet. The Pirani score improved from an average of 4.3 (range: 3-6) at presentation to a final follow-up average of 0.5 (range: 0-1). One foot had unsatisfactory result with a pretreatment score of 5.5 and a final score of 3. The results were also presented in terms of the number of casts applied, the need for tenotomy of tendo Achillis, recurrence of the deformity and the ultimate requirement for surgical release. The use of the Ponseti method in older-aged infants with idiopathic congenital clubfoot seems to be an effective method of treatment, obviating the need for extensive surgery. PMID- 19282788 TI - Radical dissection after positive groin sentinel biopsy in melanoma patients: rate of further positive nodes. AB - The aim of this retrospective study was to analyze the incidence of further nonsentinel node metastases at completion lymphadenectomy of the groin after a positive sentinel node biopsy to evaluate whether radical dissection remains the treatment of choice for these patients. Patients treated at the National Cancer Institute of Milan between January 1999 and December 2006 were reviewed retrospectively. The analysis included patients with a diagnosis of positive sentinel node biopsy of the groin (clinically negative) who underwent completion groin, iliac, and obturatory dissections. The primary melanoma was located on the lower extremities and trunk in 82.5 and 17.5%, respectively. The median follow-up was more than 30 months. The number of positive sentinel nodes was considered, as well as the size and location of the metastases (micro vs. macro). After radical dissection, the number and the location (groin, iliac, or groin+iliac nodes) of further nonsentinel node metastases were analyzed. The frequency of further nonsentinel node metastases at completion of groin dissection was correlated to Breslow's thickness and to the characteristics of the positive sentinel node biopsy. A total of 1581 patients with primary melanoma (>1 mm, or Clark's IV-V) underwent lymphatic mapping and sentinel node biopsy: 752 patients had sentinel node biopsy at the groin basin; among these, 150 (20%) patients presented positive sentinel node biopsy and underwent completion radical dissection (groin, obturatory, and external iliac+obturatory radical node dissections). We found further positive nonsentinel node metastases in 36 of 150 (24%) patients, 69% (25 of 36) of which were located in the iliac-obturator area and 31% in the groin area only: 16 patients (44.4%) had one additional metastatic node and seven patients (19.4%) had two, whereas 13 (36.1%) had three or more. In 22 cases (61.1%), the sentinel node showed a macrometastasis (>2 mm deposit in the node) and in 14 cases (38.9%) a micrometastasis (<2 mm deposit). In conclusion, there is clear evidence that patients with a positive sentinel node biopsy could have further positive nonsentinel node metastases (in our series, 24%). Although it is well known that the impact of sentinel node biopsy on survival of melanoma patients has yet to be defined, to obtain a clear nodal basin and regional control a groin+iliac-obturatory radical node dissection remains an appropriate procedure in the presence of a positive sentinel node biopsy at the groin level. This could be considered a standard treatment until new data, provided by ongoing studies, indicate new parameters for selecting patients for completion lymph node dissection. PMID- 19282789 TI - Isolated limb infusion with melphalan and dactinomycin for regional melanoma and soft-tissue sarcoma of the extremity: final report of a phase II clinical trial. AB - Isolated limb infusion (ILI) is a minimally invasive technique of delivering regional chemotherapy in patients with advanced melanoma or soft-tissue sarcoma of the limb. We report the final results of the first clinical trial of ILI in North America (NCT00004250). Eligible patients had recurrent melanoma or unresectable soft-tissue sarcoma of the limb. Angiographic catheters were positioned just above the knee or elbow of the extremity. General anesthesia was performed, a proximal tourniquet inflated, and a normothermic, low flow, hypoxic infusion of melphalan and dactinomycin circulated through the involved limb for 20 min. Tumor response and morbidity were assessed using standard criteria. Thirty-seven patients were accrued to the trial and 44 ILIs were performed (eight patients had two ILIs); one patient was not treated. Of the 32 evaluable patients, 17 (53%) had a significant response at 3 months: 25% of patients had a complete response and 28% of patients had a partial response. The median duration of complete response was 1 year (5-32 months). Morbidity was acceptable, with peak erythema, edema, and pain experienced at 2 weeks and considered 'moderate' in most patients. No patients developed compartment syndrome or required amputation because of ILI. ILI is well tolerated. More than half of the treated patients experienced a complete or partial response. PMID- 19282790 TI - Developmental expression of thyroid hormone receptor beta2 protein in cone photoreceptors in the mouse. AB - Thyroid hormone receptor beta2 (TRbeta2) controls the patterning of cone opsin photopigments that mediate colour vision. We raised an antiserum against TRbeta2 to study cone photoreceptor development by western blot and immunostaining analyses. TRbeta2-positive cells first appeared between embryonic day 10 (E10) and E12. Numbers increased until near birth, correlating with generation of the cone population. At birth, signals decreased until postnatal day 10, then declined to very low levels in adulthood. TRbeta2-positive cells were initially dispersed but became aligned at the edge of the outer neuroblastic layer by E15. Postnatally, these cells migrated inwardly until postnatal day 10, then outwardly to the edge of the outer nuclear layer, the location of mature cones. TRbeta2 represents a functionally unique marker for cone development. PMID- 19282791 TI - Usefulness of a deep-inspiration breath-hold 18F-FDG PET/CT technique in diagnosing liver, bile duct, and pancreas tumors. AB - BACKGROUND: The deep-inspiration breath-hold F-fluorodeoxyglucose PET/computed tomography (DIBH F-FDG PET/CT) technique improves the limitations of diagnosing a lesion located in an area influenced by respiratory motion that brings about spatial misregistration caused by respiration between PET and CT. However, its clinical effectiveness with regard to abdominal lesions has not been elucidated. The influence of respiratory motion for calculating the maximal standardized uptake value (SUVmax) and metabolic volume (MV) in DIBH F-FDG PET/CT has not been investigated either. OBJECTIVE: The purpose of this study was to investigate the usefulness of the DIBH F-FDG PET/CT technique in diagnosing liver tumors, bile duct cancers, and pancreas tumors. In addition, we compared the values of SUVmax and MV between DIBH and nonbreath-hold (NBH). METHODS: Forty patients with various abdominal malignancies including liver tumors, bile duct cancers, and pancreas tumors were enrolled. In total, the patients had 47 abdominal lesions. All patients showed a misregistered image in the early whole-body image taken 50 min after intravenous F-FDG infusions. We added the delayed images 40 min after the first image. On the delayed image, we carried out both conventional techniques with normal respiration (NBH) and the DIBH technique. Finally, we compared two kinds of images in each patient. At the same time, we compared both SUVmax and MV of cancer obtained by the two kinds of imaging methods. RESULTS: In 14 lesions (29.8%), we corrected the anatomical tumor location, from the incorrect to the correct organ, by the DIBH technique. In 22 lesions (46.8%), we corrected the tumor location within the organ. Consequently, tumor staging also changed in 11 patients (23.4%) after correction by the DIBH technique. Regarding the SUVmax value by DIBH, it showed an increase of approximately 15.0-58.6% compared with that measured by NBH. In contrast, the value of MV by DIBH showed a decrease of 20% compared with that measured by NBH. CONCLUSION: The DIBH F-FDG PET/CT technique is feasible for accurate localization when diagnosing of liver tumors, bile duct cancers, and pancreas cancers. The DIBH technique also improves the inaccurate quantification of both SUVmax and MV. PMID- 19282792 TI - Factors affecting the measurement of left ventricular ejection fraction in myocardial perfusion imaging. AB - BACKGROUND: Estimation of the left ventricular ejection fraction (LVEF) using myocardial perfusion imaging is increasingly used in the evaluation of coronary artery disease. This study aims to compare the effect of (i) two commercially available software packages: quantitative gated single-photon emission computed tomography (SPECT) (QGS) and 4DM-SPECT, and (ii) prereconstruction filtering, on LVEF quantification. METHODS: Images from 101 patients were reconstructed using AutoCardiac and processed using QGS and 4DM-SPECT. Filtering was performed before reconstruction using Hermes FBP SPET on a group of 32 consecutive patients using Butterworth filters (orders 5 and 10; cut-off frequency 0.5-1.2 cycles/cm). RESULTS: Good correlation was observed between QGS and 4DM-SPECT (r=0.88), with an average difference of 2.1%. The difference in LVEF between the two packages ranged from 21 to -28%. The LVEF was overestimated at cut-off frequencies < or = 0.8 cycles/cm compared with higher cut-off frequencies in 26 of 30 (87%) patients. CONCLUSION: There was a clinically significant difference between the LVEF calculated by QGS and 4DM-SPECT and consequently the two packages should not be used interchangeably. The effect of cut-off frequency on LVEF estimation was found to be very patient specific. Changing the cut-off frequency by as little as 0.1 cycles/cm can cause clinically significant differences in LVEF estimation. PMID- 19282793 TI - 99mTc-methionine scintimammography in the evaluation of breast cancer. AB - OBJECTIVE: The diagnostic utility of a C-methionine scan has been established in breast cancer. We were able to radiolabel methionine with Tc at our institute. Thus, we undertook clinical trials to determine the role of Tc-methionine scans in the detection of breast cancer. METHODS: Scintimammography was performed in 47 female (median age 44 years, range 28-68 years) patients having palpable breast masses. All of them underwent ultrasound, mammography, fine-needle aspiration cytology, and Tc-methionine scintimammography before surgery. The final diagnosis was made after histopathological examination. Tc-methionine scintimammography was done after injecting 555 MBq of radiotracer intravenously. The results of scintimammography were compared with histopathology. RESULTS: The histopathological findings were malignant in 33 (70%) and benign in 14 (30%) cases. Scintimammography showed true-positive findings in 29 patients out of 33 cases of breast cancer. True-negative findings were found in 13 out of 14 patients having benign breast lesions. The sensitivity, specificity, and positive predictive value were found to be 87.8, 92.8, and 96.6% respectively. CONCLUSION: Tc-methionine imaging can provide useful information with reasonably high sensitivity and specificity in evaluating patients having breast masses. PMID- 19282794 TI - Preparation of 166Ho-oxine-lipiodol and its preliminary bioevaluation for the potential application in therapy of liver cancer. AB - OBJECTIVE: Intra-arterial administration of beta-emitting radionuclides in the form of suitable radiopharmaceuticals is one of the promising modalities for the treatment of liver cancer. Ho [T1/2=26.9 h, Ebeta(max)=1.85 MeV, Egamma=81 keV (6.4%)] could be envisaged as an attractive radionuclide for the use in liver cancer therapy owing to its high-energy beta-emission, short half-life and feasibility of its production with adequately high specific activity and radionuclidic purity using moderate flux reactors. Lipiodol is chosen as the vehicle to deliver localized doses of ionizing radiation to liver cancer cells after intra-arterial hepatic infusion as it is selectively retained in the vascular periphery of the proliferating cells. METHODS: Ho was produced by thermal neutron bombardment on a natural Ho2O3 target at a flux of approximately 6 x 10 n/cm.s for 7 days. Radiolabelled lipiodol was prepared by dispersing the Ho-oxine complex in lipiodol. The biological behaviour of Ho-oxine-lipiodol was studied by biodistribution and imaging studies in normal Wistar rats. RESULTS: Ho was produced with a specific activity of 9.25-11.10 TBq/g and radionuclidic purity of approximately 100%. The Ho-labelled oxine complex was prepared in high yield (approximately 97%). Approximately, 95% of the Ho activity was dispersed in lipiodol within 30 min. The resulting radiolabelled preparation was found to exhibit good stability in physiological saline and human serum up to 3 days. The biodistribution and imaging studies revealed satisfactory hepatic retention (88.43+/-2.85% of injected activity after 2 days) with insignificant uptake in any other major organ/tissue except skeleton (6.44+/-1.07% at 2 days postinjection). CONCLUSION: The Ho-oxine-lipiodol preparation exhibited promising features in preliminary studies and warrants further investigation. PMID- 19282796 TI - Mortality after lumbar fusion surgery. AB - STUDY DESIGN: Retrospective population-based cohort study. OBJECTIVE: To describe mortality after lumbar fusion surgery in Washington State workers' compensation claimants in the perioperative period and beyond. SUMMARY OF BACKGROUND DATA: Although lumbar fusion surgery can be associated with serious complications, perioperative mortality is generally considered rare. Population-based mortality estimates have been limited to surgery in older adults. METHODS: We identified all Washington State workers' compensation claimants who underwent fusion between January 1994 and December 2001 (n = 2378) and assessed the frequency, timing, and causes of death. Mortality follow-up was concluded in 2004. Death was ascertained from Washington State vital statistics records and from the workers' compensation claims database. Poisson regression was used to obtain age- and gender-adjusted mortality rates. Years of potential life lost, percent of potential life lost, and mean potential life lost were calculated for the leading 5 causes of death and we calculated the risk of death associated with selected predictors. RESULTS: Among the 2378 lumbar fusion subjects in the study cohort, 103 were deceased by 2004. The 3-year cumulative mortality rate was 1.93% (95% confidence interval, 1.41%-2.57%). The 90-day perioperative mortality rate was 0.29% (95% confidence interval, 0.11%-0.60%). The risk of perioperative mortality was positively associated with repeat fusions. The age- and gender-adjusted all-cause mortality rate was 3.1 deaths per 1000 worker-years (95% confidence interval, 0.9-9.8). Analgesic-related deaths were responsible for 21% of all deaths and 31.4% of all potential life lost. The risk of analgesic-related death was higher among workers who received instrumentation or intervertebral cage devices compared with recipients of bone-only fusions (1.1% vs. 0.0%; P = 0.03) and among workers with degenerative disc disease (age- and gender-adjusted mortality rate ratio, 2.71) (95% confidence interval, 1.17-6.28). The burden was especially high among subjects between 45 and 54 years old with degenerative disc disease (rate ratio, 7.45). CONCLUSION: Analgesic-related deaths are responsible for more deaths and more potential life lost among workers who underwent lumbar fusion than any other cause. Risk of analgesic-related death was especially high among young and middle aged workers with degenerative disc disease. PMID- 19282797 TI - A long way to go: practice patterns and evidence in chronic low back pain care. AB - STUDY DESIGN: A cross-sectional, telephone survey of a representative sample of North Carolina households in 2006. OBJECTIVE: The primary objectives of these analyses were to describe health care use (providers, medications, treatments, diagnostic tests) for chronic low back pain (LBP) and relate current patterns of use to current best evidence for care of the condition. SUMMARY OF BACKGROUND DATA: Chronic LBP is common and expensive. Prior research on care utilization often was derived from medical claims databases, reflecting reimbursed health care use, often by one payer. METHODS: Five thousand three hundred fifty-seven households were contacted in 2006 to identify 732 noninstitutionalized adults 21 years and older with chronic LBP. Five hundred ninety individuals sought care. Patient reported health care utilization, comparison with efficacy was demonstrated by current systematic reviews. RESULTS: Individuals with chronic back pain were middle-aged (mean age 53 years), and the majority were women (62%). Provider and treatment use was common and varied. Sixty percent used narcotics in the previous month. The mean number of provider visits was 21, and over one-third had an advanced imaging procedure in the past year. Physical treatments were common, and often not supported by evidence. Only 3% had engaged in a formal spine rehabilitation program. Half of patients not taking antidepressants were positive on a 2-item depression screen. Although this study was population-based, it was conducted in only one state. CONCLUSION: Provider and treatment use for chronic LBP are both very common and varied. Current treatment patterns are consistent with overutilization of some medications and treatments, and underutilization of exercise and depression treatment. PMID- 19282806 TI - Ultrasound guidance can improve the outcome of botulinum toxin A injection. PMID- 19282807 TI - Game-based telerehabilitation. AB - This article summarizes the recent accomplishments and current challenges facing game-based virtual reality (VR) telerehabilitation. Specifically this article addresses accomplishments relative to realistic practice scenarios, part to whole practice, objective measurement of performance and progress, motivation, low cost, interaction devices and game design. Furthermore, a description of the current challenges facing game based telerehabilitation including the packaging, internet capabilities and access, data management, technical support, privacy protection, seizures, distance trials, scientific scrutiny and support from insurance companies. PMID- 19282808 TI - Carotid endarterectomy: results of the Italian Vascular Registry. AB - AIM: The aim of this study was to evaluate the results of carotid endarterectomy (CEA) carried out in centers dedicated to vascular surgery. METHODS: The study was supported by the Italian Registry for Vascular Activity, which collected the data of 89 centers of vascular surgery (almost all of the existing centers in Italy) during 2007. Data were collected for 5962 CEAs. A total of 5,809 patients (153 were operated bilaterally in two staged procedures), 3990 (68.7%) males and 1,819 (31.3%) females, whose ages ranged from 33 to 100 years (mean 72.7+/-7.78) underwent the surgery procedure. Several surgical techniques were used: i.e., standard CEA with direct suture (1477 cases, 24.8%), standard CEA with patch (2242 cases, 37.6%), and eversion technique (2243 cases, 37.6%). RESULTS: Combined perioperative mortality and stroke rate were also evaluated (1.1% [stroke 0.9%, mortality 0.2%]). CONCLUSIONS: These results are very good and are comparable to what has been reported in the literature during the last few years. This observation provides further proof of the effectiveness of CEA in the management of extracranial carotid disease. Moreover, these results have to be taken into account when evaluating any new therapeutic options, such as carotid stenting, before accepting them as valid alternatives. PMID- 19282810 TI - The role of procalcitonin as predictor for neurological deficits after carotid endarterectomy. AB - AIM: Outcome of carotid endarterectomy (CEA) is defined by mortality rate as well as the neurological outcome due to cerebral ischemia. Thus the aim of this study was to evaluate the role of the acute phase protein procalcitonin (PCT) as a predictor for neurological deficits after carotid endarterectomy. METHODS: Fifty five patients with high grade stenosis of the internal carotid artery and interdisciplinary consensus for endarterectomy were followed. Neurological examination was performed before and after the procedure to analyze perioperative neurological deficits. Blood samples were obtained before and after CEA and procalcitonin was analyzed in 55 consecutive patients (65.5% symptomatic/34.5% asymptomatic). RESULTS: No perioperative or in-hospital death was observed. Major complications did not occur, two patients suffered from bleeding requiring surgical intervention and one patient had a temporary peripheral facial nerve lesion. Postoperative neurological examination revealed no new deficit, there was no significant change of PCT (level pre- and post-CEA (the mean preoperative PCT was 0.25 ng/mL [SD 0.78, min 0.1, max 4.3]; the mean postoperative PCT was 0.11 ng/mL [SD 0.06, min 0.1, max 0.5]). There was no association found between perioperative neurological deficit and PCT. CONCLUSIONS: The present study demonstrates that there is still not sufficient evidence to recommend PCT measurement as a predictor for perioperative neurological deficit during CEA. PMID- 19282811 TI - Endovascular proximal control of ruptured abdominal aortic aneurysms: the internal aortic clamp. AB - Ruptured abdominal aortic aneurysm (RAAA) is the most common and devastating complication affecting a patient with abdominal aortic aneurysm (AAA). Despite advances in surgery and critical care, the mortality rate associated with RAAA remains largely unchanged. Emergency open repair is the gold standard conventional treatment of RAAA but is associated with a high mortality rate. The physiologic challenges associated with general anaesthetic induction such as loss of the sympathetic vasoconstrictor tone with consequent hypotension, and the anatomic challenges associated with external aortic cross-clamping such as calcification, friability, or poor visualisation of the aneurysm neck, have led to the adoption of endovascular techniques in the surgical treatment of RAAA. Promising results of endovascular repair of ruptured abdominal aortic aneurysm (REVAR) have been reported. In addition, the provision of endovascular aortic control by inflating a compliant aortic occlusion balloon (AOB) proximal to the ruptured aneurysm, as an internal aortic clamp, has been successfully used in haemodynamically unstable patients undergoing either REVAR or emergency open repair of RAAA. An AOB is inserted under local anaesthesia and can be introduced through either the transbrachial or the transfemoral routes, each with its own advantages and disadvantages. This review aimed at providing an up-to-date overview of the current knowledge concerning endovascular proximal aortic control using an AOB with emphasis on the rationale, position, benefits, and drawbacks of its use. PMID- 19282812 TI - Incidental detection by [11C]choline PET/CT of meningiomas in prostate cancer patients. AB - AIM: Anti-androgenic hormonal therapy in prostate cancer patients with concomitant meningioma may result in tumor growth and development of neurological symptoms. Positron emission tomography/computed tomography (PET/CT) with [11C]choline is used for restaging prostate cancer patients with biochemical failure. In vitro and in vivo data support altered choline metabolism in meningiomas. METHODS: During a retrospective study in prostate cancer patients with biochemical failure referred to our institution between November 2004 and January 2007, encephalic focal uptake of [11C]choline was incidentally noted in 4 patients, 2 of which had been taking luteinizing hormone-releasing hormone analogs. RESULTS: Subsequent to the incidental finding, one patient underwent surgical removal of the meningioma; strict neuroradiological follow-up was planned for the 3 other patients. CONCLUSION: We suggest that in prostate cancer patients candidate for anti-androgenic therapy the whole body [11C]choline PET/CT scan should include the whole skull to check for the possible presence of meningiomas. This could help to identify patients at risk for the development of neurological symptoms during anti-androgenic therapy and help the referring urologist in the clinical management of these patients. PMID- 19282813 TI - State-specific prevalence and trends in adult cigarette smoking--United States, 1998-2007. AB - Cigarette smoking in the United States results in an estimated 443,000 premature deaths and $193 billion in direct health-care expenditures and productivity losses each year. During 2007, an estimated 19.8% of adults in the United States were current smokers. To update 2006 state-specific estimates of cigarette smoking, CDC analyzed data from the 2007 Behavioral Risk Factor Surveillance System (BRFSS) survey and examined trends in cigarette smoking from 1998-2007. Results of these analyses indicated substantial variation in current cigarette smoking during 2007 (range: 8.7%-31.1%) among the 50 states, the District of Columbia (DC), Guam, Puerto Rico (PR), and the U.S. Virgin Islands (USVI). Trend analyses of 1998-2007 data indicated that smoking prevalence decreased in 44 states, DC, and PR, and six states had no substantial changes in prevalence after controlling for age, sex, and race/ethnicity. However, only Utah and USVI met the Healthy People 2010 target for reducing adult smoking prevalence to 12% (objective 27-1a). The Institute of Medicine (IOM) calls for full implementation of comprehensive, evidence-based tobacco control programs at CDC-recommended funding levels to achieve substantial reductions in tobacco use in all states and areas. PMID- 19282814 TI - Investigation of a genotype cluster of tuberculosis cases--Detroit, Michigan, 2004-2007. AB - In August 2007, the Detroit Department of Health and Wellness Promotion, Michigan Department of Community Health (MDCH), and CDC investigated a genotype cluster of eight tuberculosis (TB) cases in U.S.- born patients in the Detroit metropolitan area. The cases had been reported during December 2004-April 2007. The first case was reported in a patient (the index patient) whose drug-susceptible TB subsequently developed multidrug resistance. Seven additional cases were reported in patients with Mycobacterium tuberculosis genotypes that matched the genotype of the index patient. These included one case of multidrug-resistant (MDR) TB in a young relative of the index patient and one case in the index patient's parent, who died from TB meningitis. This report describes the investigation and illustrates the importance of ensuring that each case of TB disease is promptly recognized and successfully treated and that all close contacts of TB patients are identified, evaluated, and treated for latent TB infection if indicated. PMID- 19282815 TI - Simian malaria in a U.S. traveler--New York, 2008. AB - Four species of intraerythrocytic protozoa of the genus Plasmodium (P. falciparum, P. vivax, P. ovale, and P. malariae) are known to cause malaria in humans. However, recent reports from Asia suggest the possibility that a fifth malaria species, Plasmodium knowlesi, is emerging as an important zoonotic human pathogen. Although more than 20 species of Plasmodium can infect nonhuman primates, until recently, naturally acquired human infections of simian malaria were viewed as rare events lacking public health significance. When viewed by light microscopy (the gold standard for laboratory diagnosis of malaria), many of the simian species are almost indistinguishable from the four Plasmodium species that cause infection in humans. Molecular techniques, such as polymerase chain reaction (PCR) amplification and microsatellite analysis, are needed for definitive species determination. This report describes the first recognized case of imported simian malaria in several decades in the United States, diagnosed in 2008 in a patient from New York who had traveled to the Philippines. Atypical features of the parasite seen on light microscopy triggered further molecular testing, which confirmed the diagnosis of P. knowlesi. To date, all simian malaria species have been susceptible to chloroquine treatment. Molecular analysis of certain malaria parasites isolated from ill travelers returning to the United States from Asia or South America can more accurately assess the burden of simian malaria parasite infections in humans. PMID- 19282816 TI - Plasma levels of inflammatory markers neopterin, sialic acid, and C-reactive protein in pregnancy and preeclampsia. AB - BACKGROUND: To determine whether the cellular inflammatory marker of activated macrophages and monocytes, neopterin (NEO), and the acute-phase inflammatory markers sialic acid (SA) and C-reactive protein (CRP) are elevated in pregnancy and further elevated in the pregnancy syndrome preeclampsia. METHODS: Maternal plasma concentrations of NEO, SA, and CRP were measured by high-sensitivity enzyme-linked immunosorbent assay (ELISA) or high-performance liquid chromatography in 20 nonpregnant women, 40 women with uncomplicated pregnancies, 50 women with transient hypertension of pregnancy alone, 49 women with small for gestational age (SGA) infants without preeclampsia, and 47 women with preeclampsia. RESULTS: The mean concentration of plasma NEO, SA, and CRP were all significantly elevated in all groups of pregnant women compared to nonpregnant women (P < 0.001 for all). In addition, maternal plasma NEO concentrations were further elevated in women with preeclampsia compared to the other groups of pregnant women (P < 0.01). As expected, the acute-phase inflammatory markers CRP and SA correlated positively with each other. However, CRP was also correlated with the activated macrophage and monocyte marker NEO in women with transient hypertension of pregnancy and with preeclampsia (P < 0.05). CONCLUSIONS: The inflammatory markers NEO, SA, and CRP are all elevated during pregnancy. However, only NEO, a marker of macrophage and monocyte activation, was further elevated in women with preeclampsia. These data suggest that there is a striking increase in inflammation during pregnancy, and cellular immune activation is further elevated during preeclampsia. PMID- 19282817 TI - A study on polymorphisms of elastin gene in Chinese Han patients with isolated systolic hypertension. AB - BACKGROUND: Elastin (ELN) is mainly located in the internal elastic lamina of large arteries. Degradation of ELN is expected to induce large vessel stiffness, which could lead to elderly systolic hypertension. Recent studies have shown that polymorphism of ELN is associated with stiffness of elastic arteries and elevated blood pressure; however, there are no further studies on isolated systolic hypertension (ISH). METHODS: We identified the genotype of the ELN gene in 358 patients with ISH, 413 essential hypertension (EH) patients with elevated diastolic blood pressure (DBP), and 244 age-matched normotensive (NT) controls for five single-nucleotide polymorphisms (SNPs) and detected the brachial-ankle pulse wave velocity (baPWV), C-reactive protein (CRP), and intima-media thickness (IMT) for these patients. RESULTS: ISH was statistically significant in association with SNP rs34208922 (A allele frequency was 0.068 in ISH patients, 0.036 in EH patients, and 0.014 in NT controls; P < 0.001, P(corr) < 0.005) and possibly with SNP rs2071307 (A allele frequency was 0.103 in ISH patients, 0.079 in EH patients, and 0.047 in NT controls; P = 0.002, P(corr) = 0.01), however, the A allele frequency was not different between ISH patients and EH patients. In addition, baPWV and CRP were significantly associated with SNP rs34208922 and rs2071307. The other three SNPs were not significantly associated with ISH, baPWV, CRP, or IMT. Haplotypes of TGGTA and TGAT- were also significantly associated with ISH (P = 0.0001, P(corr) = 0.0021; P = 0.0023, P(corr) = 0.0483). CONCLUSIONS: Variants within the ELN gene are associated with increased risk of ISH and aortic stiffness in the Chinese Han population. PMID- 19282818 TI - In-hospital mortality among black patients admitted for hypertension-related disorders in Mbuji Mayi, Congo. AB - BACKGROUND: As a consequence of hypertension, sub-Saharan Africa faces an epidemic of cardiovascular disease. METHODS: We assessed in-hospital mortality among patients admitted for hypertension-related diseases to two city hospitals in Mbuji Mayi, Congo. On admission, we obtained anthropometric measurements, blood pressure, information on cardiovascular risk factors, and on the awareness and treatment of hypertension. We modeled the probability of death, using stepwise logistic regression. RESULTS: Of 401 consecutive patients (mean age, 54.3 years; 129 women), 118 (29.4 %) were unaware of their hypertension. Among 283 aware patients (70.6%), 126 (44.5%) were untreated. Systolic/diastolic blood pressure on admission averaged 178/106 mm Hg. In addition to hypertension, 390 patients (97.3%) had other cardiovascular risk factors, including a creatinine clearance below 60 ml/min/1.73 m(2) in 47 subjects (11.7%). Over 15 days (median) of hospitalization, 89 deaths (22.2%) occurred. The multivariable-adjusted probability of death increased with systolic blood pressure (odds ratio (95% confidence interval) for +10 mm Hg, 1.43 (1.15-1.77); P < 0.01), body mass index (+5 kg/m(2), 2.40 (1.39-4.17); P < 0.01), being aware vs. unaware of hypertension (3.17 (1.52-6.61); P < 0.01), and being untreated (2.33 (1.12-4.76); P < 0.05), but it decreased with age (+10 years, 0.65 (0.46-0.92); P < 0.05) and higher creatinine clearance (+10 ml/min/1.73 m(2), 0.71 (0.61-0.82); P < 0.001). CONCLUSIONS: The in-hospital mortality among African patients hospitalized for hypertension-related disorders in a Congolese provincial capital city is over 20%. These findings underscore that screening and treatment for hypertension and the prevention of cardiovascular disease should be placed much higher on the political agenda in sub-Saharan Africa. PMID- 19282819 TI - Visceral and subcutaneous adiposity and brachial artery vasodilator function. AB - Endothelial dysfunction may link obesity to cardiovascular disease (CVD). We tested the hypothesis that visceral abdominal tissue (VAT) as compared with subcutaneous adipose tissue (SAT) is more related to endothelium-dependent vasodilation. Among Framingham Offspring and Third Generation cohorts (n = 3,020, mean age 50 years, 47% women), we used multivariable linear regression adjusted for CVD and its risk factors to relate computed tomography (CT)-assessed VAT and SAT, BMI, and waist circumference (WC), with brachial artery measures. In multivariable-adjusted models, BMI, WC, VAT, and SAT were positively related to baseline artery diameter and baseline mean flow velocity (all P < 0.001), but not hyperemic mean flow velocity. In multivariable-adjusted models, BMI (P = 0.002), WC (P = 0.001), and VAT (P = 0.01), but not SAT (P = 0.24) were inversely associated with percentage of flow-mediated dilation (FMD%). However, there was little incremental increase in the proportion of variability explained by VAT (R(2) = 0.266) as compared to SAT (R(2) = 0.265), above and beyond traditional risk factors. VAT, but not SAT was associated with FMD% after adjusting for clinical covariates. Nevertheless, the differential association with VAT as compared to SAT was minimal. PMID- 19282820 TI - Diabetes of the liver: the link between nonalcoholic fatty liver disease and HFCS 55. AB - Nonalcoholic fatty liver disease (NAFLD) is associated with obesity and insulin resistance. It is also a predisposing factor for type 2 diabetes. Dietary factors are believed to contribute to all three diseases. NAFLD is characterized by increased intrahepatic fat and mitochondrial dysfunction, and its etiology may be attributed to excessive fructose intake. Consumption of high fructose corn syrup 55 (HFCS-55) stands at up to 15% of the average total daily energy intake in the United States, and is linked to weight gain and obesity. The aim of this study was to establish whether HFCS-55 could contribute to the pathogenesis of NAFLD, by examining the effects of HFCS-55 on hepatocyte lipogenesis, insulin signaling, and cellular function, in vitro and in vivo. Exposure of hepatocytes to HFCS-55 caused a significant increase in hepatocellular triglyceride (TG) and lipogenic proteins. Basal production of reactive oxygen metabolite (ROM) was increased, together with a decreased capacity to respond to an oxidative challenge. HFCS-55 induced a downregulation of the insulin signaling pathway, as indicated by attenuated (ser473)phosphorylation of AKT1. The c-Jun amino-terminal kinase (JNK), which is intimately linked to insulin resistance, was also activated; and this was accompanied by an increase in endoplasmic reticulum (ER) stress and intracellular free calcium perturbation. Hepatocytes exposed to HFCS-55 exhibited mitochondrial dysfunction and released cytochrome C (CytC) into the cytosol. Hepatic steatosis and mitochondrial disruption was induced in vivo by a diet enriched with 20% HFCS 55; accompanied by hypoadiponectinemia and elevated fasting serum insulin and retinol-binding protein-4 (RBP4) levels. Taken together our findings indicate a potential mechanism by which HFCS-55 may contribute to the pathogenesis of NAFLD. PMID- 19282821 TI - Dopamine for "wanting" and opioids for "liking": a comparison of obese adults with and without binge eating. AB - Obesity research suffers from an overinclusion paradigm whereby all participants with a BMI beyond a certain cutoff value (e.g., 30) are typically combined in a single group and compared to those of normal weight. There has been little attempt to identify meaningful subgroups defined by their salient biobehavioral differences. In order to address this limitation, we examined genetic and psychological indicators of hedonic eating in obese adults with (n=66) and without (n=70) binge eating disorder (BED). Our analyses focused on dopamine (DA) and opioid genetic markers because of their conjoint association with the functioning of brain reward mechanisms. We targeted three functional polymorphisms related to the D2 receptor (DRD2) gene, as well as the functional A118G polymorphism of the mu-opioid receptor (OPRM1) gene. We found that significantly more obese controls had the "loss-of-function" A1 allele of Taq1A compared to their BED counterparts, whereas the "gain-of-function" G allele of A118G occurred with greater frequency in the BED group. A significant gene-gene combination chi2 analysis also indicated that of those participants with the gain gain genotype (G+ and A1), 80% were in the BED group whereas only 35% with the loss-loss genotype (G- and A1+) were in this group. Finally, BED subjects had significantly higher scores on a self-report measure of hedonic eating. Our findings suggest that BED is a biologically based subtype of obesity and that the proneness to binge eating may be influenced by a hyper-reactivity to the hedonic properties of food--a predisposition that is easily exploited in our current environment with its highly visible and easily accessible surfeit of sweet and fatty foods. PMID- 19282822 TI - Voluntary exercise and its effects on body composition depend on genetic selection history. AB - Little is known about how genetic variation affects the capacity for exercise to change body composition. We examined the extent to which voluntary exercise alters body composition in several lines of selectively bred mice compared to controls. Lines studied included high runner (HR) (selected for high wheel running), M16 (selected for rapid weight gain), Institute of Cancer Research (ICR) (randomly bred as control for M16), M16i (an inbred line derived from M16), HE (selected for high percentage of body fat while holding body weight constant), LF (selected for low percentage of body fat), C57BL/6J (common inbred line), and the F1 between HR and C57BL/6J. Body weight and body fat were recorded before and after 6 days of free access to running wheels in males and females that were individually caged. Total food intake was measured during this 6-day period. All pre- and postexercise measures showed significant strain effects. While HR mice predictably exercised at higher levels, all other selection lines had decreased levels of wheel running relative to ICR. The HR x B6 F1 ran at similar levels to HR demonstrating complete dominance for voluntary exercise. Also, all strains lost body fat after exercise, but the relationships between exercise and changes in percent body were not uniform across genotypes. These results indicate that there is significant genetic variation for voluntary exercise and its effects on body composition. It is important to carefully consider genetic background and/or selection history when using mice to model effects of exercise on body composition, and perhaps, other complex traits as well. PMID- 19282823 TI - Obesity and television watching in preschoolers in Greece: the GENESIS study. AB - The aim of the current work was to evaluate the effect of preschoolers' television (TV) watching time on the prevalence of obesity even after controlling for their total energy intake and their physical activity status. A representative sample of 2,374 Greek children aged 1-5 years was examined ("Growth, Exercise and Nutrition Epidemiological Study in preSchoolers", GENESIS study). Children's TV watching time on a usual weekday and at a usual weekend was recorded. The overall mean of children's TV viewing time was 1.32 h/day. The majority of participants (74.0%) spent <2 h/day watching TV whereas only 3.1% spent >4 h/day in front of a TV set. Overall, 65.2% of participants were normal weight, 17.2% were overweight, and the rest 17.6% were obese. The prevalence of obesity was significantly higher among those with TV viewing time >or=2 h/day (21.7%) compared to those watching TV <2 h/day (16.1%, P = 0.003). TV viewing time remained significantly associated with the likelihood of being obese even after controlling for potential confounders (i.e., socio demographic and other characteristics and physical activity status) only among children aged 3-5 years. However, further adjusting for children's total energy intake revealed that the association between the TV viewing time and the probability of being obese was no longer statistically significant. On the other hand, physical activity status continued to be an independent factor of being obese. The current findings support the hypothesis that the effect of TV viewing time on childhood obesity is independent of physical activity status and may be attributed to the increased total energy intake during TV watching. PMID- 19282824 TI - Family physicians' practices and attitudes regarding care of extremely obese patients. AB - Despite the growing epidemic of extreme obesity in the United States, weight management is not adequately addressed in primary care. This study assessed family physicians' practices and attitudes regarding care of extremely obese patients and factors associated with them. A cross-sectional, self-administered survey was mailed to 500 family physicians in New Jersey (NJ) during March-May 2008. Measures included knowledge, weight management approaches, attitudes toward managing obesity, challenges with examinations, availability of supplies, and strategies to improve care. Response rate was 53% (N = 255). Bariatric surgery and weight loss medications were infrequently recommended, particularly in physicians with higher volume of extremely obese patients (odds ratio (OR) 0.38; 95% confidence interval (CI) 0.23, 0.62 and OR 0.51; 95% CI 0.31, 0.85 for surgery and medications, respectively). Higher knowledge was associated with increased frequency of recommendations of weight loss medications (P < 0.0001) and bariatric surgery (P < 0.0001). There was a high prevalence of negative attitudes, particularly in younger physicians and those with lower patient volume. Increased knowledge of weight-loss diets was associated with less dislike in discussing weight loss (P < 0.0001), less frustration (P = 0.0001), less belief that treatment is often ineffective (P < 0.0001), and less pessimism about patient success (P = 0.0002). Many providers encountered challenges performing examinations on extremely obese patients. More education of primary care physicians, particularly on bariatric surgery, specific examination techniques, and availability of community resources for obese persons is needed. Further research is needed to determine if interventions to increase knowledge of physicians will lead to less negative attitudes toward weight loss and extremely obese patients. PMID- 19282825 TI - Rise in extracellular fluid volume during high sodium depends on BMI in healthy men. AB - A high sodium (HS) intake is associated to increased cardiovascular and renal risk, especially in overweight subjects. We hypothesized that abnormal sodium and fluid handling is involved, independent of hypertension or insulin resistance. Therefore, we studied the relation between BMI and sodium-induced changes in extracellular fluid volume (ECFV; distribution volume of (125)I-iothalamate) in 78 healthy men, not selected for BMI. A total of 78 subjects with a median BMI of 22.5 (range: 19.2-33.9 kg/m(2)) were studied after 1 week on a low sodium (LS) diet (50 mmol Na(+)/d) and after 1 week on HS (200 mmol Na(+)/d). The change from LS to HS resulted in an increase in ECFV of 1.2 +/- 1.8 l. Individual changes in ECFV were correlated to BMI (r = 0.361, P < 0.01). Furthermore, in response to HS, a higher BMI was associated to a higher rise in filtered load of sodium (FL(Na(+)) = [Na(+)] x GFR, r = 0.281, P < 0.05). Thus, a shift to HS leads to a larger rise in ECFV in healthy subjects with higher BMI, associated with an elevated FL(Na(+)) during HS. Although no hypertension occurred in these healthy subjects, our data provide a potential explanation for the interaction of sodium intake and BMI on cardiovascular and renal risk. Exaggerated fluid retention may be an early pathogenic factor in the cardiorenal complications of overweight. PMID- 19282826 TI - Cardiorespiratory fitness and the risk of overweight in youth: the Healthy Hearts Longitudinal Study of Cardiometabolic Health. AB - The primary objective of this longitudinal study was to determine the association between cardiorespiratory fitness and the risk of overweight status in youth. To accomplish this aim we analyzed data from annual school-based surveys of cardiorespiratory fitness and anthropometry conducted between 2004 and 2006. The first analysis was performed on a cohort of 902 youth aged 6-15 years followed for 12 months to assess the association between cardiorespiratory fitness levels determined from a graded maximal field test and the risk of becoming overweight. The second analysis was conducted on a cohort of 222 youth followed for 2 years to assess the continuous association between annual changes fitness and weight gain. Children with low cardiorespiratory fitness were characterized by higher waist circumference and disproportionate weight gain over the 12-month follow-up period (P < 0.05). Within the entire cohort, the 12-month risk of overweight classification was 3.5-fold (95% confidence = 2.0-6.0, P < 0.001) higher in youth with low cardiorespiratory fitness, relative to fit peers. A time series mixed effects regression model revealed that reductions in cardiorespiratory fitness were significantly and independently associated with increasing BMI (r = -0.18, P < 0.05) in youth. Accordingly, low cardiorespiratory fitness and reductions in fitness over time are significantly associated with weight gain and the risk of overweight in children 6-15 years old. An assessment of cardiorespiratory fitness using a common field test may prove useful for the identification of youth at risk of overweight and serve as a potential target for obesity prevention. PMID- 19282827 TI - Maternal feeding practices become more controlling after and not before excessive rates of weight gain. AB - It is unclear whether controlling maternal feeding practices (CMFPs) lead to or are a response to increases in a child's BMI. Our goal was to determine the direction of this relationship. Data were obtained from National Institute of Child Health and Human Development's Study of Early Child Care and Youth Development. Child BMI z-score (zBMI) was calculated from measured weight and height. CMFP was defined by, "Do you let your child eat what he/she feels like eating?". Change in child zBMI was calculated between 4-7 years and 7-9 years, and dichotomized into "increasing" vs. "no change or decreasing". Change in CMFP was calculated over the same time periods, and dichotomized into "more controlling" vs. "no change or less controlling." Multiple logistic regression, stratified by gender and controlling for race, maternal education, maternal weight status, and baseline child weight status, was used for analysis. A total of 789 children were included. From 4 to 9 years, mean zBMI increased (P = 0.02) and mothers became more controlling (P < 0.001). Increasing CMFP between 4 and 7 years was associated with decreased odds of increasing zBMI between 7 and 9 years in boys (odds ratio = 0.52, 95% confidence interval = 0.27-1.00). There was no relationship in girls. Increasing zBMI between 4 and 7 years was associated with increasing CMFPs between 7 and 9 years in girls (odds ratio = 1.72, 95% confidence interval = 1.08-2.74), but not boys. Early increases in CMFP were not associated with later increases in zBMI for boys or girls. However, early increases in zBMI among girls were associated with later increases in CMFP. Clarifying the relationship between maternal feeding practices and child weight will inform future recommendations. PMID- 19282828 TI - Differential responses of orexigenic neuropeptides to fasting in offspring of obese mothers. AB - Maternal obesity due to long-term high-fat diet (HFD) consumption leads to faster growth in offspring during suckling, and increased adiposity at 20 days of age. Decreased expression of the orexigenic neuropeptide Y (NPY) and increased anorexigenic proopiomelanocortin (POMC) mRNA expression were observed in the fed state. However, hunger is the major drive to eat and hypothalamic appetite regulators change in response to meals. Therefore, it is important to compare both satiated and fasting states. Female Sprague-Dawley rats (8 weeks old) were fed a cafeteria-style HFD (15.33 kJ/g) or chow for 5 weeks before mating, with the same diet continuing throughout gestation and lactation. At postnatal day 20, male pups were killed either after overnight fasting or in the fed state. Pups from obese dams were hyperphagic during both pre- and postweaning periods. Pups from obese dams had higher hypothalamic mRNA expression of POMC and NPY Y1 receptor, but lower hypothalamic melanocortin-4 receptor (MC4R) and its downstream target single-minded gene 1 (Sim1), in the fed state. Overnight fasting reduced circulating glucose, insulin, and leptin and increased hypothalamic NPY Y1 receptor mRNA in pups from both lean and obese dams. Hypothalamic NPY and agouti-related protein (AgRP) were only increased by fasting in pups from obese dams; reductions in MC4R and Sim1 were only seen in pups from lean dams. At weaning, the suppressed orexigenic signals in offspring from obese dams were normalized after overnight fasting, although anorexigenic signaling appeared impaired in these animals. This may contribute to their hyperphagia and faster growth. PMID- 19282829 TI - Estimating activity-related energy expenditure under sedentary conditions using a tri-axial seismic accelerometer. AB - Activity-related energy expenditure (AEE) is difficult to quantify, especially under sedentary conditions. Here, a model was developed using the detected type of physical activity (PA) and movement intensity (MI), based on a tri-axial seismic accelerometer (DynaPort MiniMod; McRoberts B.V., The Hague, the Netherlands), with energy expenditure for PA as a reference. The relation between AEE (J/min/kg), MI, and the type of PA was determined for standardized PAs as performed in a laboratory including: lying, sitting, standing, and walking. AEE (J/min/kg) was calculated from total energy expenditure (TEE) and sleeping metabolic rate (SMR) as assessed with indirect calorimetry ((TEEx0.9)-SMR). Subsequently, the model was validated over 23-h intervals in a respiration chamber. Subjects were 15 healthy women (age: 22+/-2 years; BMI: 24.0+/-4.0 kg/m2). Predicted AEE in the chamber was significantly related to measured AEE both within (r2=0.81+/-0.06, P<0.00001) and between (r2=0.70, P<0.001) subjects. The explained variation in AEE by the model was higher than the explained variation by MI alone. This shows that a tri-axial seismic accelerometer is a valid tool for estimating AEE under sedentary conditions. PMID- 19282830 TI - RUNX1 mutations are frequent in chronic myelomonocytic leukemia and mutations at the C-terminal region might predict acute myeloid leukemia transformation. AB - Runt-related transcription factor 1 (RUNX1) is essential for normal hematopoiesis. RUNX1 mutations have rarely been reported in chronic myelomonocytic leukemia (CMML). We examined RUNX1 mutations in 81 patients with CMML at initial diagnosis. Mutational analysis was performed on bone marrow samples by direct sequencing of all reverse transcription PCR products amplified with three primer pairs that cover the entire coding sequences of RUNX1b. Thirty two RUNX1 mutations were detected in 30 patients (37%); 23 mutants were located in the N-terminal part and 9 in the C-terminal region. The mutations consisted of 9 missense, 1 silent, 7 nonsense and 15 frameshift mutations. Two patients had biallelic heterozygous mutations. There was no difference in overall survival between patients with and without RUNX1 mutations, but a trend of higher risk of acute myeloid leukemia (AML) progression was observed in mutation-positive patients (16/30 vs 17/51, P=0.102), especially in patients with C-terminal mutations (P=0.023). The median time to AML progression was 6.8 months in patients with C-terminal mutations compared with 28.3 months in those without mutations (P=0.022). This study showed for the first time a high frequency of RUNX1 mutations in CMML. C-terminal mutations might be associated with a more frequent and rapid AML transformation. PMID- 19282831 TI - The proteasome inhibitor bortezomib interacts synergistically with the histone deacetylase inhibitor suberoylanilide hydroxamic acid to induce T leukemia/lymphoma cells apoptosis. AB - Interactions between inhibitors of the proteasome and histone deacetylases have been examined in human T-leukemia/lymphoma cells both in vitro and in vivo. Co exposure of cells to bortezomib and suberoylanilide hydroxamic acid (SAHA) synergistically induces T-leukemia/lymphoma cells to undergo apoptosis, consistent with a significant increase in mitochondrial injury and caspase activation. These events are accompanied by inhibition of cyto-protective signaling pathways, including the nuclear factor (NF)-kappaB, Raf-1/mitogen induced extracellular kinase (MEK)/extracellular signal-related kinase (ERK) and AKT pathways, and activation of stress-related cascades, including the stress activated kinases c-jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK). Moreover, bortezomib in conjunction with SAHA efficiently induces apoptosis of primary T-leukemia/lymphoma cells and inhibits tumor growth in a murine xenograft model established with subcutaneous injection of Jurkat cells. Taken together, these findings confirm the synergistic anti-tumor effect of the proteasome and histone deacetylase inhibitors, and provide an insight into the future clinical applications of bortezomib-SAHA combining regimen in treating T-cell malignancies. PMID- 19282832 TI - Continuous Fli-1 expression plays an essential role in the proliferation and survival of F-MuLV-induced erythroleukemia and human erythroleukemia. AB - Erythroleukemia induced by Friend Murine Leukemia Virus (F-MuLV) serves as a powerful tool for the study of multistage carcinogenesis and hematological malignancies in mice. Fli-1, a proto-oncogene and member of the Ets family, is activated through viral integration in F-MuLV-induced erythroleukemia, and is the most critical event in the induction of this disease. Fli-1 aberrant regulation is also observed in human malignancies, including Ewing's sarcoma, which is often linked to expression of the EWS/Fli-1 fusion oncoprotein. Here we examined the effects of Fli-1 inhibition to further elucidate its role in these pathological occurrences. The constitutive suppression of Fli-1, through RNA interference (RNAi), inhibits growth and induces death in F-MuLV-induced erythroleukemia cells. Expression of a dominant negative protein Engrailed (En)/Fli-1 reduces proliferation of EWS/Fli-1-transformed NIH-3T3 cells, and both F-MuLV-induced and human erythroleukemia cells. F-MuLV-induced erythroleukemia cells also display increased apoptosis, associated with reduced expression of bcl-2, a known fli-1 target gene. Introduction of En/Fli-1 into an F-MuLV-infected erythroblastic cell line induces differentiation, as shown by increased alpha-globin expression. These results suggest, for the first time, an essential role for continuous Fli-1 overexpression in the maintenance and survival of the malignant phenotype in murine and human erythroleukemias. PMID- 19282833 TI - Six-year follow-up of patients receiving imatinib for the first-line treatment of chronic myeloid leukemia. AB - Imatinib mesylate is considered standard of care for first-line treatment of chronic phase chronic myeloid leukemia (CML-CP). In the phase III, randomized, open-label International Randomized Study of Interferon vs STI571 (IRIS) trial, previously untreated CML-CP patients were randomized to imatinib (n=553) or interferon-alpha (IFN) plus cytarabine (n=553). This 6-year update focuses on patients randomized to receive imatinib as first-line therapy for newly diagnosed CML-CP. During the sixth year of study treatment, there were no reports of disease progression to accelerated phase (AP) or blast crisis (BC). The toxicity profile was unchanged. The cumulative best complete cytogenetic response (CCyR) rate was 82%; 63% of all patients randomized to receive imatinib and still on study treatment showed CCyR at last assessment. The estimated event-free survival at 6 years was 83%, and the estimated rate of freedom from progression to AP and BC was 93%. The estimated overall survival was 88% -- or 95% when only CML related deaths were considered. This 6-year update of IRIS underscores the efficacy and safety of imatinib as first-line therapy for patients with CML. PMID- 19282834 TI - Altered naive and memory CD4+ T-cell homeostasis and immunosenescence characterize younger patients with myelodysplastic syndrome. AB - Response to immunosuppressive therapy (IST) in younger patients with myelodysplastic syndrome (MDS) has been linked to a T-cell-dominant autoimmune process that impairs hematopoiesis. Analysis of the age-adjusted CD4:CD8 ratio in 76 MDS patients compared with 54 healthy controls showed that inadequate CD4+, rather than expansion of CD8+ T cells, was associated with a lower ratio in a group that included both lower and higher risk MDS patients defined by the International Prognostic Scoring System. In younger MDS patients, naive and memory phenotypes defined by CD45RA and CD62L display showed depletion of naive CD4+ and CD8+ T cells, suggesting a possible relationship to IST responsiveness. To determine the correlation between T-cell subset distribution, T-cell turnover and autoimmunity, a cohort of 20 patients were studied before and after IST. The CD4:CD8 ratio correlated inversely with the proliferative T-cell index before treatment in IST-responsive patients, suggesting that proliferation may be linked to accelerated CD4+ T-cell turnover and hematopoietic failure. Our data show seminal findings that both CD4+ and CD8+ T-cell subsets are dysregulated in MDS. Association between these T-cell defects and response to IST suggests that aberrant T-cell homeostasis and chronic activation are critical determinants influencing autoimmune hematopoietic suppression in younger patients. PMID- 19282835 TI - Increased risk for CNS relapse in pre-B cell leukemia with the t(1;19)/TCF3-PBX1. AB - To evaluate the impact of contemporary therapy on the clinical outcome of children with pre-B acute lymphoblastic leukemia (ALL) and the t(1;19)/TCF3/PBX1, we analyzed 735 patients with B-cell precursor ALL treated in four successive protocols at St Jude Children's Research Hospital. The 41 patients with the t(1;19) had a comparable event-free survival to that of the 694 patients with other B-cell precursor ALL (P=0.63; 84.2+/-7.1% (s.e.) vs 84.0+/-1.8% at 5 years). However, patients with the t(1;19) had a lower cumulative incidence of any hematological relapse (P=0.06; 0 vs 8.3+/-1.2% at 5 years) but a significantly higher incidence of central nervous system (CNS) relapse (P<0.001; 9.0+/-5.1% vs 1.0+/-0.4% at 5 years). In a multivariate analysis, the t(1;19) was an independent risk factor for isolated CNS relapse. These data suggest that with contemporary treatment, patients with the t(1;19) and TCF3/PBX1 fusion have a favorable overall outcome but increased risk of CNS relapse. PMID- 19282836 TI - Non-neuronal expression of transient receptor potential type A1 (TRPA1) in human skin. PMID- 19282837 TI - Bcl-3 acts as an innate immune modulator by controlling antimicrobial responses in keratinocytes. AB - Innate immune responses involve the production of antimicrobial peptides (AMPs), chemokines, and cytokines. We report here the identification of B-cell leukemia (Bcl)-3 as a modulator of innate immune signaling in keratinocytes. In this study, it is shown that Bcl-3 is inducible by the Th2 cytokines IL-4 and IL-13 and is overexpressed in lesional skin of atopic dermatitis (AD) patients. Bcl-3 was shown to be important to cutaneous innate immune responses as silencing of Bcl-3 by small-interfering RNA (siRNA) reversed the downregulatory effect of IL-4 on the HBD3 expression. Bcl-3 silencing enhanced vitamin D3 (1,25D3)-induced gene expression of cathelicidin AMP in keratinocytes, suggesting a negative regulatory function on cathelicidin transcription. Furthermore, 1,25D3 suppressed Bcl-3 expression in vitro and in vivo. This study identified Bcl-3 as an important modulator of cutaneous innate immune responses and its possible therapeutic role in AD. PMID- 19282838 TI - Mechanism of sustained release of vascular endothelial growth factor in accelerating experimental diabetic healing. AB - In this study, we hypothesize that local sustained release of vascular endothelial growth factor (VEGF), using adenovirus vector (ADV)-mediated gene transfer, accelerates experimental wound healing. This hypothesis was tested by determining the specific effects of VEGF(165) application on multiple aspects of the wound healing process, that is, time to complete wound closure and skin biomechanical properties. After showing accelerated wound healing in vivo, we studied the mechanism to explain the findings on multiple aspects of the wound healing cascade, including epithelialization, collagen deposition, and cell migration. Intradermal treatment of wounds in non-obese diabetic and db/db mice with ADV/VEGF(165) improves healing by enhancing tensile stiffness and/or increasing epithelialization and collagen deposition, as well as by decreasing time to wound closure. VEGF(165), in vitro, stimulates the migration of cultured human keratinocytes and fibroblasts, thus revealing a non-angiogenic effect of VEGF on wound closure. In conclusion, ADV/VEGF is effective in accelerating wound closure by stimulating angiogenesis, epithelialization, and collagen deposition. In the future, local administration and sustained, controlled release of VEGF(165) may decrease amputations in patients with diabetic foot ulcers and possibly accelerate closure of venous ulcers and pressure ulcers. PMID- 19282839 TI - Rituximab mediates a strong elevation of B-cell-activating factor associated with increased pathogen-specific IgG but not autoantibodies in pemphigus vulgaris. AB - Pemphigus vulgaris (PV) is a severe autoimmune disease affecting the skin and mucous membranes, characterized by autoantibodies mainly against desmoglein 3 (dsg3). This study investigated the effects of different treatment options on two B-cell mediators, B-cell-activating factor (BAFF) and a proliferation-inducing ligand (APRIL), in 19 PV patients on immunosuppressive drugs alone or in combination with immunoadsorption and anti-CD20 antibody, respectively. Serum BAFF and APRIL levels, circulating desmoglein-reactive autoantibodies, and serum IgG specific for varicella-zoster virus (VZV) and Epstein-Barr virus (EBV) were determined by ELISA before and at different time points after initiation of the respective therapy. In contrast to immunosuppressive therapy alone and in combination with adjuvant immunoadsorption, respectively, rituximab treatment led to a strong and significant elevation of BAFF, but not of APRIL levels, which normalized upon recovery of peripheral CD19(+) B cells. Moreover, rituximab treatment led to a statistically significant increase of anti-VZV-IgG and anti EBV-IgG titers, whereas anti-dsg1 and -3 specific autoantibody titers decreased significantly. Our results suggest that elevated BAFF levels might exert a differential effect on the induction of autoreactive versus pathogen-specific IgG antibody production in PV patients, possibly due to promotion of antibody release of pathogen-specific long-lived plasma cells. PMID- 19282840 TI - The unfolded protein response is activated in differentiating epidermal keratinocytes. AB - The unfolded protein response (UPR), which is induced by stress to the endoplasmic reticulum (ER), is involved in the functional alteration of certain cells, such as the differentiation of B cells to plasma cells. The aim of this study is to determine whether the UPR is activated during epidermal keratinocyte (KC) differentiation. Here, we show that the expression of the UPR-induced proteins Bip/GRP78 and HRD1 was increased in cells in the supra-basal layers of normal human epidermis that contain KCs undergoing differentiation as well as in skin-equivalent cultured KCs. However, Bip/GRP78 and HRD1 were poorly expressed in proliferating KCs in squamous cell carcinoma and psoriasis vulgaris tissues. The epidermal growth factor receptor tyrosine kinase inhibitor, PD153035, which induces KC differentiation, upregulated UPR-induced marker mRNAs and proteins. Furthermore, microarray analyses and quantitative PCR revealed that ER stress inducing reagents, tunicamycin (TU), thapsigargin, and brefeldin A, altered the expression of genes essential for human epidermal KC differentiation, including C/EBPbeta, KLF4, and ABCA12 in vitro. However, ABCA12 and KLF4 mRNA did not increase with TU treatment after siRNA-mediated knockdown of XBP-1. Taken together, our findings strongly suggest that the UPR is activated during normal epidermal KC differentiation and induces C/EBPbeta, KLF4, and ABCA12 mRNAs. PMID- 19282841 TI - Prevalence, severity, and severity risk factors of acne in high school pupils: a community-based study. AB - A cross-sectional, community-based study was performed to determine the prevalence and severity of acne vulgaris in adolescents and of factors influencing the acne severity risk. The presence of acne was clinically determined and the secondary outcome measures of family acne history and the relation of acne to nutrition habits, emotional stress, menstruation, and smoking were recorded in a questionnaire. A representative sample of 1,002 pupils aged 16+/-0.9 years was enrolled. The overall acne prevalence was 93.3, 94.4% for boys and 92.0% for girls. Moderate to severe acne was observed in 14%. The prevalence of moderate to severe acne was 19.9% in pupils with and 9.8% in those without a family history of acne (P<0.0005; OR: 2.3). Acne severity risk increased with the number of family members with acne history. A mother with acne history influenced the severity of acne the most. Increasing pubertal age, seborrhea, the premenstrual phase, mental stress, and sweet and oily foods were recognized as risk factors for moderate to severe acne. In contrast, gender, spicy foods, and smoking were not associated with acne severity. In conclusion, acne is a common disorder in Iranian adolescents, with a low rate of moderate to severe acne. A genetic background is suggested, with mother's acne history being the most important prognostic factor. Skin quality and certain nutrition habits may affect acne severity. PMID- 19282842 TI - Involvement of Propionibacterium acnes in the augmentation of lipogenesis in hamster sebaceous glands in vivo and in vitro. AB - Propionibacterium acnes is considered to be involved in the aggravation of acne vulgaris, but it remains unclear whether P. acnes directly influences lipogenesis in sebaceous glands. In this study, we showed that a culture medium of P. acnes (acnes-CM) and formalin-killed P. acnes (F-acnes) prepared from P. acnes strains, JCM6473 and JCM6425, intracellularly augmented lipid droplet formation and triacylglycerol (TG) synthesis in undifferentiated and insulin-differentiated hamster sebocytes. Acnes-CM and F-acnes prepared from four clinical P. acnes strains elicited the same lipogenesis augmentation. The augmented TG production resulted from an increase in the diacylglycerol acyltransferase activity. Topical application of acnes-CM to the skin of hamster auricles every day for 4 weeks revealed that sebum accumulation was augmented in sebaceous glands and ducts. Furthermore, both acnes-CM and F-acnes increased the production of 15-deoxy Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a cytochrome P450 (CYP)-linked sebaceous lipogenic factor, in differentiated sebocytes. A CYP inhibitor, SKF 525A, decreased the acnes-CM- and F-acnes-augmented production of TG and 15d PGJ(2). Thus, to our knowledge these results provide previously unreported evidence that P. acnes directly participates in the augmentation of sebaceous lipogenesis through a proposed mechanism in which an increase of 15d-PGJ(2) production through the CYP pathway is closely associated with the enhancement of TG production. PMID- 19282843 TI - Pathogenic anti-desmoglein MAbs show variable ELISA activity because of preferential binding of mature versus proprotein isoforms of desmoglein 3. PMID- 19282844 TI - Evaluation of the clonal origin of multiple primary melanomas using molecular profiling. AB - Numerous investigations have been conducted using molecular profiling to evaluate the possible clonal origin of second malignancies in various cancer types. However, to date no study assessing clonality of multiple primaries has been conducted in melanoma. In this investigation using patients treated at a specialist melanoma treatment center, we compared the somatic mutational profiles of pairs of melanomas designated as independent on the basis of thorough assessment of their clinical and pathologic characteristics. We used a set of highly polymorphic genetic markers selected on the basis of their chromosomal positions and the frequencies of reported allelic losses at these genetic loci. Our statistical testing strategy showed no significant evidence of clonal origin of the two primaries in 17 of the 19 patients examined. The results suggest that most second melanomas designated as independent primary tumors on the basis of their clinicopathologic features are indeed independent occurrences of the disease, supporting the validity of the criteria used by experienced pathologists in distinguishing new primaries from metastases. PMID- 19282845 TI - Discrepancies between the fate of myoblast xenograft in mouse leg muscle and NMR label persistency after loading with Gd-DTPA or SPIOs. AB - 1H-NMR (nuclear magnetic resonance) imaging is regularly proposed to non invasively monitor cell therapy protocols. Prior to transplantation, cells must be loaded with an NMR contrast agent (CA). Most studies performed so far make use of superparamagnetic iron oxide particles (SPIOs), mainly for favorable detection sensitivity. However, in the case of labeled cell death, SPIO recapture by inflammatory cells might introduce severe bias. We investigated whether NMR signal changes induced by preloading with SPIOs or the low molecular weight gadolinium (Gd)-DTPA accurately monitored the outcome of transplanted cells in a murine model of acute immunologic rejection. CA-loaded human myoblasts were grafted in the tibialis anterior of C57BL/6 mice. NMR imaging was repeated regularly until 3 months post-transplantation. Label outcome was evaluated by the size of the labeled area and its relative contrast to surrounding tissue. In parallel, immunohistochemistry assessed the presence of human cells. Data analysis revealed that CA-induced signal changes did not strictly reflect the graft status. Gd-DTPA label disappeared rapidly yet with a 2-week delay compared with immunohistochemical evaluation. More problematically, SPIO label was still visible after 3 months, grossly overestimating cell survival (<1 week). SPIOs should be used with extreme caution to evaluate the presence of grafted cells in vivo and could hardly be recommended for the long-term monitoring of cell transplantation protocols. PMID- 19282846 TI - Real-time monitoring of cell transplantation in mouse dystrophic muscles by a secreted alkaline phosphatase reporter gene. AB - Transplantation of muscle precursor cells (MPCs) is a promising approach for the treatment of muscular dystrophies. However, preclinical and clinical results have shown that the technology is not yet efficient enough for most therapeutic applications. Among the problems that remain unsolved are low cellular survival, poor proliferation and lack of migration of the transplanted cells. One major technical hurdle for the optimization of transplantation protocols is how to follow precisely the fate of the cells after transplantation. In this study, we examined the use of a secreted form of the mouse alkaline phosphatase (mSeAP) enzyme as the reporter system transduced into MPCs using a retroviral vector. We show that circulating mSeAP could be detected in the serum of the transplanted mice at different time points after MPC transplantation. We also found that the level of circulating mSeAP is highly correlated with the number of transplanted cells and that mSeAP is an excellent histological marker. Further, studying the levels of circulating mSeAP compared with the number of muscle fibers positive to mSeAP and to dystrophin, enabled detailed analyses of bottleneck steps for successful transplantation. Taken together, our results show that mSeAP is an excellent quantitative 'real-time' reporter gene for cell therapy preclinical studies. PMID- 19282847 TI - Dendritic cells and T cells deliver oncolytic reovirus for tumour killing despite pre-existing anti-viral immunity. AB - Reovirus is a naturally occurring oncolytic virus currently in early clinical trials. However, the rapid induction of neutralizing antibodies represents a major obstacle to successful systemic delivery. This study addresses, for the first time, the ability of cellular carriers in the form of T cells and dendritic cells (DC) to protect reovirus from systemic neutralization. In addition, the ability of these cellular carriers to manipulate the subsequent balance of anti viral versus anti-tumour immune response is explored. Reovirus, either neat or loaded onto DC or T cells, was delivered intravenously into reovirus-naive or reovirus-immune C57Bl/6 mice bearing lymph node B16tk melanoma metastases. Three and 10 days after treatment, reovirus delivery, carrier cell trafficking, metastatic clearance and priming of anti-tumour/anti-viral immunity were assessed. In naive mice, reovirus delivered either neat or through cell carriage was detectable in the tumour-draining lymph nodes 3 days after treatment, though complete clearance of metastases was only obtained when the virus was delivered on T cells or mature DC (mDC); neat reovirus or loaded immature DC (iDC) gave only partial early tumour clearance. Furthermore, only T cells carrying reovirus generated anti-tumour immune responses and long-term tumour clearance; reovirus loaded DC, in contrast, generated only an anti-viral immune response. In reovirus immune mice, however, the results were different. Neat reovirus was completely ineffective as a therapy, whereas mDC--though not iDC--as well as T cells, effectively delivered reovirus to melanoma in vivo for therapy and anti-tumour immune priming. Moreover, mDC were more effective than T cells over a range of viral loads. These data show that systemically administered neat reovirus is not optimal for therapy, and that DC may be an appropriate vehicle for carriage of significant levels of reovirus to tumours. The pre-existing immune status against the virus is critical in determining the balance between anti-viral and anti tumour immunity elicited when reovirus is delivered by cell carriage, and the viral dose and mode of delivery, as well as the immune status of patients, may profoundly affect the success of any clinical anti-tumour viral therapy. These findings are therefore of direct translational relevance for the future design of clinical trials. PMID- 19282848 TI - Braf(V600E) cooperates with Pten loss to induce metastatic melanoma. AB - Mutational activation of BRAF is the earliest and most common genetic alteration in human melanoma. To build a model of human melanoma, we generated mice with conditional melanocyte-specific expression of BRaf(V600E). Upon induction of BRaf(V600E) expression, mice developed benign melanocytic hyperplasias that failed to progress to melanoma over 15-20 months. By contrast, expression of BRaf(V600E) combined with Pten tumor suppressor gene silencing elicited development of melanoma with 100% penetrance, short latency and with metastases observed in lymph nodes and lungs. Melanoma was prevented by inhibitors of mTorc1 (rapamycin) or MEK1/2 (PD325901) but, upon cessation of drug administration, mice developed melanoma, indicating the presence of long-lived melanoma-initiating cells in this system. Notably, combined treatment with rapamycin and PD325901 led to shrinkage of established melanomas. These mice, engineered with a common genetic profile to human melanoma, provide a system to study melanoma's cardinal feature of metastasis and for preclinical evaluation of agents designed to prevent or treat metastatic disease. PMID- 19282849 TI - Measurement of auxin transport in Arabidopsis thaliana. AB - This protocol allows the measurement of auxin transport in roots, hypocotyls and inflorescences of Arabidopsis thaliana plants by examining transport of radiolabeled auxin or movement of an auxin-induced gene expression signal. The protocol contains four stages: seedling growth, auxin application, a transport period of variable length, and quantification of auxin movement or reporter expression. Beyond the time for plant growth, the transport assay can be completed within 4-18 h. Auxin is applied to seedlings in agar cylinders or droplets, which does not require specialized liquid-handling equipment or micromanipulators, in contrast with methods that apply auxin in liquid droplets. Spatial control of auxin application is reduced, but this method has the advantages of being technically more feasible for most laboratories and allowing agar containing radioactive auxin to be removed for pulse chase assays that determine transport rates. These methods allow investigation of genetic and environmental factors that control auxin transport. PMID- 19282850 TI - Labeling of biotin antibodies with horseradish peroxidase using cyanuric chloride. AB - In this report, we describe a two-step protocol for labeling of an affinity purified antibody to biotin with horseradish peroxidase (HRP) using cyanuric chloride (CC) as a bridge. The enzyme was first modified with CC, and following chromatography on a PD-10 column, the activated HRP was incubated with the antibody to effect coupling of the two proteins. Assessment of the conjugate product was carried out using ELISA and SDS-PAGE electrophoresis where evidence for high antibody activity, high specific activity of the conjugate preparation, coupling of nearly all the antibody and over 90% of the enzyme was shown. The titer of the conjugate exceeded 1/100,000. High molecular weight complexes were observed in the SDS-PAGE results, indicating an efficient conjugation procedure. The presence of high molecular weight complexes indicated an efficient conjugation procedure. The protocol is simple, and the conjugation steps can be completed in 27 h once the preparatory phase has been carried out; the method is entirely generic and may be applied to labeling of any antibody. PMID- 19282851 TI - The role of IFN-gamma Elispot assay in HIV vaccine research. AB - The interferon (IFN)-gamma Elispot assay has been widely used as a general screening method for the quantification and characterization of the human immunodeficiency virus (HIV)-specific CD8+ T cell responses. However, the predictive power of this assay has been challenged due to the lack of efficacy of a recently conducted HIV vaccine phase IIb trial, despite induction of robust Elispot responses. This finding plus improvements in multiparameter flow cytometry, which has the potential advantage of simultaneously quantifying numerous parameters, raises questions regarding the future role of IFN-gamma Elispot as a gateway to moving forward with clinical trials of candidate vaccines. However, the IFN-gamma Elispot assay has been, unlike other techniques, evaluated and validated in several proficiency panels and is advantageous in cost effectively detecting and mapping T-cell responses. Here we present a detailed protocol for a state-of-the-art 3-d IFN-gamma Elispot assay and review further advantages and disadvantages of this method for the characterization of HIV specific CD8+ T cell responses. PMID- 19282852 TI - Trophic macrophages in development and disease. AB - Specialized phagocytes are found in the most primitive multicellular organisms. Their roles in homeostasis and in distinguishing self from non-self have evolved with the complexity of organisms and their immune systems. Equally important, but often overlooked, are the roles of macrophages in tissue development. As discussed in this Review, these include functions in branching morphogenesis, neuronal patterning, angiogenesis, bone morphogenesis and the generation of adipose tissue. In each case, macrophage depletion impairs the formation of the tissue and compromises its function. I argue that in several diseases, the unrestrained acquisition of these developmental macrophage functions exacerbates pathology. For example, macrophages enhance tumour progression and metastasis by affecting tumour-cell migration and invasion, as well as angiogenesis. PMID- 19282853 TI - Models of haematopoiesis: seeing the wood for the trees. AB - Many models of haematopoiesis dictate that haematopoietic stem cells make an early and irrevocable choice between lymphoid and myeloid cell fates. However, recent data show that intermediate progenitors have both lymphoid and myeloid potential, thereby refuting early theories and leading to a lack of consensus in the field. In this Opinion article we present a simple pairwise relationships model of cell fate determination that does not depend on the underlying developmental branch points that in other models dictate a preferred route (or routes) to a particular cell fate. PMID- 19282856 TI - Specific podocin mutations determine age of onset of nephrotic syndrome all the way into adult life. AB - In steroid-resistant nephrotic syndrome (SRNS) Machuca et al. report that mutations of the recessive podocin gene cause adult-onset SRNS if the R229Q genetic variant occurs in a compound heterozygous state with another podocin mutation. Learning to tell apart the specific allele combinations of podocin mutations will be important for prognosis, genetic counseling in living related kidney donation, accurate etiologic classification within treatment studies, and the understanding of podocin function. PMID- 19282857 TI - Periodontal disease: a modifiable risk factor for cardiovascular disease in ESRD patients? AB - Accumulating evidence suggests that periodontal disease is associated with increased risk of cardiovascular disease (CVD). Several mechanisms have been proposed to explain this association. To date, however, a causal relation has not been firmly established. In addition, the extent to which treatment of periodontal disease might result in lower incidence of CVD has not been addressed in any study to date. Further research is needed before recommendations are made regarding monitoring and therapy of periodontal disease. PMID- 19282858 TI - Neutrophils in acute kidney injury: not neutral any more. AB - Awad and colleagues elucidate the spatiotemporal profile of neutrophil infiltration in the kidney following ischemia-reperfusion injury. Using elegant in vivo labeling techniques, they demonstrate increased neutrophil content in the kidney following ischemia-reperfusion, which is largely due to transmigration from the circulation into the interstitial compartment. The authors also provide mechanistic insights into this phenomenon and show that adenosine 2A receptor agonists reduce interstitial neutrophil infiltration and improve renal function. PMID- 19282860 TI - Critical confounders in the prognostic role of cellular biomarkers. PMID- 19282861 TI - The case. A suicidal woman with delayed high anion gap metabolic acidosis. PMID- 19282862 TI - Tunneled dialysis catheter tip migration in obese and large-breasted individuals. PMID- 19282863 TI - Association of candidate gene polymorphisms with chronic kidney disease in Japanese individuals with hypertension. AB - Although hypertension has been recognized as a risk factor for chronic kidney disease (CKD), the genetic factors for predisposition to CKD in individuals with hypertension remain largely unknown. The purpose of this study was to identify the genetic variants that confer susceptibility to CKD among individuals with hypertension. The study population comprised 3696 Japanese individuals with hypertension (2265 men, 1431 women), including 1257 individuals (789 men, 468 women) with CKD (estimated glomerular filtration rate (eGFR) <60 ml min(-1) per 1.73 m(2)) and 2439 controls (1476 men, 963 women; eGFR >or=60 ml min(-1) per 1.73 m(2)). The genotypes for 30 polymorphisms of 26 candidate genes were determined. An initial screening of allele frequencies by the chi(2)-test revealed that eight polymorphisms were significantly (false discovery rate <0.05) associated with the prevalence of CKD in hypertensive individuals. Subsequent multivariable logistic regression analysis with adjustment for covariates as well as a stepwise forward selection procedure revealed that the T --> C (Val591Ala) polymorphism of APOB (rs679899), the -681C --> G polymorphism of PPARG (rs10865710), the T --> C (Cys1367Arg) polymorphism of WRN (rs1346044), the -850C --> T polymorphism of TNF (rs1799724), the -219G --> T polymorphism of APOE (rs405509), the C --> T polymorphism of PTGS1 (rs883484) and the 41A --> G (Glu14Gly) polymorphism of ACAT2 (rs9658625) were significantly (P<0.05) associated with the prevalence of CKD. Our results suggest that APOB, WRN, ACAT2, APOE, PPARG, TNF and PTGS1 are susceptibility loci for CKD among Japanese individuals with hypertension. Determination of the genotypes for these polymorphisms may prove informative for the assessment of genetic risk for CKD among such individuals. PMID- 19282864 TI - Coronary flow reserve: a new target for treating hypertension. PMID- 19282865 TI - Association of polymorphisms of SORBS1, GCK and WISP1 with hypertension in community-dwelling Japanese individuals. AB - Although various loci and genes have been implicated in predisposition to hypertension by genetic linkage analyses and candidate gene association studies, the genes that confer susceptibility to this condition remain to be identified definitively. We have now examined the relationships of 22 candidate gene polymorphisms with the prevalence of hypertension and with blood pressure (BP) in a 6-year population-based longitudinal cohort study and observed significant relationships of three polymorphisms of SORBS1, GCK and WISP1 with hypertension. The 2233 subjects (1106 women, 1127 men) were aged 40-79 years and were randomly recruited to a population-based prospective cohort study of aging and age-related diseases in Japan. BP was measured with subjects having rested in the sitting position for at least 15 min. Genotypes for the 682A --> G (Thr228Ala) polymorphism of SORBS1, the -30G --> A polymorphism of GCK and the 2364A --> G polymorphism of WISP1 were determined by melting curve analysis. Longitudinal analysis with a generalized estimating equation revealed that the polymorphisms of SORBS1 and GCK and that of WISP1 were significantly associated with the prevalence of hypertension in women and men, respectively. Longitudinal analysis with a mixed-effect model revealed that the polymorphism of SORBS1 was significantly related to diastolic BP in women and that those of GCK and WISP1 were significantly related to both systolic and diastolic BP in women and men, respectively. These results suggest that SORBS1 and GCK are susceptibility loci for hypertension in Japanese women and that WISP1 is such a locus in men. PMID- 19282867 TI - Molecular stages of rapid and uniform neuralization of human embryonic stem cells. AB - Insights into early human development are fundamental for our understanding of human biology. Efficient differentiation of human embryonic stem cells (hESCs) into neural precursor cells is critical for future cell-based therapies. Here, using defined conditions, we characterized a new method for rapid and uniform differentiation of hESCs into committed neural precursor cells (designated C NPCs). Dynamic gene expression analysis identified several distinct stages of ESC neuralization and revealed functional modules of coregulated genes and pathways. The first wave of gene expression changes, likely corresponding to the transition through primitive ectoderm, started at day 3, preceding the formation of columnar neuroepithelial rosettes. The second wave started at day 5, coinciding with the formation of rosettes. The majority of C-NPCs were positive for both anterior and posterior markers of developing neuroepithelium. In culture, C-NPCs became electrophysiologically functional neurons; on transplantation into neonatal mouse brains, C-NPCs integrated into the cortex and olfactory bulb, acquiring appropriate neuronal morphologies and markers. Compared to rosette-NPCs,(1) C NPCs exhibited limited in vitro expansion capacity and did not express potent oncogenes such as PLAG1 or RSPO3. Concordantly, we never detected tumors or excessive neural proliferation after transplantation of C-NPCs into mouse brains. In conclusion, our study provides a framework for future analysis of molecular signaling during ESC neuralization. PMID- 19282868 TI - The Pirh2-keratin 8/18 interaction modulates the cellular distribution of mitochondria and UV-induced apoptosis. AB - Intermediate filaments (IFs) provide crucial structural support in higher eukaryotic cells. Accumulating evidences show that IFs also participate in various cellular activities including stress responses, cell growth, cell death and cell migration through dynamic interactions with various non-structure proteins. Here we report the identification of Pirh2, a RING-H2-type ubiquitin E3 ligase, as a novel binding partner of the cytoplasmic IF proteins keratin 8/18 (K8/18). Phosphorylation of either Pirh2 or K8/18 affects their association. Although Pirh2 was not found to influence the stability of K8/18, it displayed an unexpected role in regulating the organization of the network of K8/18 keratin filaments. Disruption of Pirh2-K8/K18 interaction by either UV irradiation or knockdown with Pirh2 or K18 led to the aggregation of K8/18 keratin filaments. It further induced mitochondrial redistribution, and this process is likely through a microtubule-mediated pathway. The abnormal localization of mitochondria in Pirh2-knockdown cells may partially account for its increased cell sensitivity to UV-induced apoptosis, probably through enhancing the release of pro-apoptotic proteins, such as cytochrome c and Smac/DIABLO to the cytosol. Overall, our data reveal the novel role of the Pirh2-K8/18 complex in governing the distribution of mitochondria. PMID- 19282869 TI - Drosophila HtrA2 is dispensable for apoptosis but acts downstream of PINK1 independently from Parkin. AB - High temperature requirement A2 (HtrA2/Omi) is a mitochondrial protease that exhibits proapoptotic and cell-protective properties and has been linked to Parkinson's disease (PD). Impaired mitochondrial function is a common trait in PD patients, and is likely to play a significant role in pathogenesis of parkinsonism, but the molecular mechanisms remain poorly understood. Genetic studies in Drosophila have provided valuable insight into the function of other PD-linked genes, in particular PINK1 and parkin, and their role in maintaining mitochondrial integrity. Recently, HtrA2 was shown to be phosphorylated in a PINK1-dependent manner, suggesting it might act in the PINK1 pathway. Here, we describe the characterization of mutations in Drosophila HtrA2, and genetic analysis of its function with PINK1 and parkin. Interestingly, we find HtrA2 appears to be dispensable for developmental or stress-induced apoptosis. In addition, we found HtrA2 mutants share some phenotypic similarities with parkin and PINK1 mutants, suggesting that it may function in maintaining mitochondrial integrity. Our genetic interaction studies, including analysis of double-mutant combinations and epistasis experiments, suggest HtrA2 acts downstream of PINK1 but in a pathway parallel to Parkin. PMID- 19282870 TI - Isolation of ORCTL3 in a novel genetic screen for tumor-specific apoptosis inducers. AB - We have established a systematic high-throughput screen for genes that cause cell death specifically in transformed tumor cells. In a first round of screening, cDNAs that induce apoptosis in a transformed human cell line are detected. Positive genes are subsequently tested in a synthetic lethal screen in normal cells versus their isogenic counterparts that have been transformed by a particular oncogene. In this way, the organic cation transporter-like 3 (ORCTL3) gene was found to be inactive in normal rat kidney (NRK) cells, but to induce apoptosis in NRK cells transformed by oncogenic H-ras. ORCTL3 also causes cell death in v-src-transformed cells and in various human tumor cell lines but not in normal cells or untransformed cell lines. Although ORCTL3 is a member of the organic cation transporter gene family, our data indicate that this gene induces apoptosis independently of its putative transporter activity. Rather, various lines of evidence suggest that ORCTL3 brings about apoptosis by an endoplasmic reticulum stress-mediated mechanism. Finally, we detected ORCTL3 to be downregulated in human kidney tumors. PMID- 19282871 TI - Synaptojanin-1 plays a key role in astrogliogenesis: possible relevance for Down's syndrome. AB - There is increasing interest in gliogenesis as the relevance of glia to both brain development and pathology becomes better understood. However, little is known about this process. The use of multidimensional protein identification technology (MudPIT) to identify changes in phosphoprotein levels in rat neural precursor cells treated with cytokines or retinoic acid showed that phosphorylation of the catalytic subunit of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K p110alpha) and dephosphorylation of the inositol phosphatase synaptojanin-1 were common to the gliogenic stimuli. Although PI3K was found to be involved in both neuro- and astrogliogenesis, synaptojanin-1 was specifically involved in astrogliogenesis of neural precursor cells. The role of synaptojanin 1 in astrogliogenesis was further confirmed by analysis of neuron- and glia specific markers in synaptojanin-1 knockout mouse brain. Additional experiments showed that the Sac1-like phosphatase domain of synaptojanin-1 is responsible for the observed astrogliogenic effect. Our results strongly indicate that phosphatidylinositol metabolism plays a key role in astrogliogenesis. The relevance of our findings for Down's syndrome pathology is discussed. PMID- 19282872 TI - William's syndrome: gene expression is related to parental origin and regional coordinate control. AB - William's syndrome (WS) features a spectrum of neurocognitive and behavioral abnormalities due to a rare 1.5 MB deletion that includes about 24-28 genes on chromosome band 7q11.23. Study of the expression of these genes from the single normal copy provides an opportunity to elucidate the genetic and epigenetic controls on these genes as well as their roles in both WS and normal brain development and function. We used quantitative RT-PCR to determine the transcriptional level of 14 WS gene markers in a cohort of 77 persons with WS and 48 normal controls. Results reported here: (1) show that the expression of the genes deleted in WS is decreased in some but not all cases, (2) demonstrate that the parental origin of the deletion contributes to the level of expression of GTF2I independently of age and gender and (3) indicate that the correlation of expression between GTF2I and some other genes in the WS region differs in WS subjects and normal controls, which in turn points toward a regulatory role for this gene. Interspecies comparisons suggest GTF2I may play a key role in normal brain development. PMID- 19282873 TI - Genetic insights into the origins of Tibeto-Burman populations in the Himalayas. AB - The Himalayan mountain range has played a dual role in shaping the genetic landscape of the region by (1) delineating east-west migrations including the Silk Road and (2) restricting human dispersals, especially from the Indian subcontinent into the Tibetan plateau. In this study, 15 hypervariable autosomal STR loci were employed to evaluate the genetic relationships of three populations from Nepal (Kathmandu, Newar and Tamang) and a general collection from Tibet. These Himalayan groups were compared to geographically targeted worldwide populations as well as Tibeto-Burman (TB) speaking groups from Northeast India. Our results suggest a Northeast Asian origin for the Himalayan populations with subsequent gene flow from South Asia into the Kathmandu valley and the Newar population, corroborating a previous Y-chromosome study. In contrast, Tamang and Tibet exhibit limited genetic contributions from South Asia, possibly due to the orographic obstacle presented by the Himalayan massif. The TB groups from Northeast India are genetically distinct compared to their counterparts from the Himalayas probably resulting from prolonged isolation and/or founder effects. PMID- 19282874 TI - A pharmacogenetic study of CD4 recovery in response to HIV antiretroviral therapy in two South African population groups. AB - South Africa, like many other Southern African countries, has one of the highest HIV infection rates in the world and many individuals consequently receive antiretroviral therapy (ART). However, knowledge regarding (i) the prevalence of functional single nucleotide polymorphisms (SNPs) in pharmacologically relevant genes, and (ii) variance in pharmacotherapy both within and between different populations and ethnic groups is limited. The aim of this study was to determine whether selected polymorphisms in cytochrome P450 (CYP) genes (CYP2B6 and CYP3A4) and the multidrug-resistance 1 (ABCB1) gene underlie altered antiretroviral (ARV) drug response in two South African populations. DNA samples from 182 HIV-positive individuals of Mixed-Ancestry and Xhosa ethnicity on ART were genotyped for the A 392G SNP in CYP3A4, the G516T and A785G SNPs in CYP2B6, and the T-129C, C1236T, G2677T/A and C3435T SNPs in ABCB1. Univariate two-way analysis of variance (ANOVA) testing revealed no apparent effect of ethnicity on immune recovery (in terms of CD4-cell count) in response to ART. Univariate one-way ANOVA testing revealed a discernible effect of genotype on immune recovery in the cases of the T-129C (P=0.03) and G2677A (P<0.01) polymorphisms in the ABCB1 gene. This study serves as a basis for better understanding and possible prediction of pharmacogenetic risk profiles and drug response in individuals and ethnic groups in South Africa. PMID- 19282875 TI - Replication studies for the association of PSMA6 polymorphism with coronary artery disease in East Asian populations. AB - Coronary artery disease (CAD) has become a major health problem in many countries because of its increasing prevalence and high mortality. Recently, an association of a functional sequence variation, -8C>G, in the human proteasome subunit alpha type 6 gene (PSMA6) with the susceptibility to CAD was reported. To validate the association, we investigated a total of 1330 cases and 2554 controls from Japanese and Korean populations for PSMA6 genotypes, and no evidence of the association was obtained in both Japanese (odds ratio (OR)=1.03, 95% confidence interval (CI); 0.90-1.19, P=0.66, allele count model) and Korean populations (OR=1.00, 95% CI; 0.86-1.17, P=0.95, allele count model). However, when a meta analysis of data from this study and previously reported six replication studies was done, OR was 1.08 for the G allele (95% CI; 1.02-1.14, P=0.0057), suggesting that the contribution of PSMA6 to CAD was not large enough to be readily replicated. Further studies are required to establish the contribution of this variant in the susceptibility to CAD. PMID- 19282876 TI - New atpenins, NBRI23477 A and B, inhibit the growth of human prostate cancer cells. AB - The growth and metastasis of prostate cancer are regulated by prostate stroma through the tumor-stromal cell interactions. Small molecules that modulate the tumor-stromal cell interactions will be new anticancer drugs. In the course of our screening of the modulators, we isolated two new atpenins, NBRI23477 A (4) and B (5), from the fermentation broth of Penicillium atramentosum PF1420. Compounds 4 and 5 as well as atpenin A4 (1), A5 (2) and B (3) inhibited the growth of human prostate cancer DU-145 cells in the coculture with human prostate stromal cells more strongly than that of DU-145 cells alone. PMID- 19282877 TI - A novel macrolide compound from Streptomyces bingchenggensis: fermentation, isolation, structure elucidation and biological properties. PMID- 19282878 TI - New sesquiterpenes, JBIR-27 and -28, isolated from a tunicate-derived fungus, Penicillium sp. SS080624SCf1. AB - In the course of our screening program for novel metabolites from tunicate derived fungi, novel sesquiterpenoids, named JBIR-27 (1) and -28 (2), together with known sporogen-AO1 and phomenone, were isolated from the culture broth of Penicillium sp. SS080624SCf1. The structures of 1 and 2 were determined to be eremophilane analogs on the basis of extensive NMR and MS analyses. Sporogen-AO1, phomenone and 2 showed cytotoxicity against human cervical carcinoma cell line HeLa at IC(50) values of 8.3, 19 and 92 microM, respectively, whereas 1 was inactive at a concentration of 80 microM. PMID- 19282879 TI - Pennicitrinone D, a new citrinin dimer from the halotolerant fungus Penicillium notatum B-52. PMID- 19282881 TI - Comment on 'When in Rome, reform'. PMID- 19282882 TI - In the land of the rising sun with the COP9 signalosome and related Zomes. Symposium on the COP9 signalosome, Proteasome and eIF3. PMID- 19282883 TI - Change we can believe in--of the conformational type. Workshop on the Initiation of Antigen Receptor Signaling. PMID- 19282884 TI - Human U1 snRNA forms a new chromatin-associated snRNP with TAF15. AB - The U1 small nuclear RNA (snRNA)--in the form of the U1 spliceosomal Sm small nuclear ribonucleoprotein particle (snRNP) that contains seven Sm and three U1 specific RNP proteins-has a crucial function in the recognition and removal of pre-messenger RNA introns. Here, we show that a fraction of human U1 snRNA specifically associates with the nuclear RNA-binding protein TBP-associated factor 15 (TAF15). We show that none of the known protein components of the spliceosomal U1-Sm snRNP interacts with the newly identified U1-TAF15 snRNP. In addition, the U1-TAF15 snRNP tightly associates with chromatin in an RNA dependent manner and accumulates in nucleolar caps upon transcriptional inhibition. The Sm-binding motif of U1 snRNA is essential for the biogenesis of both U1-Sm and U1-TAF15 snRNPs, suggesting that the U1-TAF15 particle is produced by remodelling of the U1-Sm snRNP. A demonstration that human U1 snRNA forms at least two structurally distinct snRNPs supports the idea that the U1 snRNA has many nuclear functions. PMID- 19282885 TI - [The state gets the physicians it deserves]. PMID- 19282886 TI - [Prescription shopping]. PMID- 19282887 TI - [Prescription shopping of addictive drugs in Norway]. AB - BACKGROUND: A Regular General Practitioner Scheme was introduced in Norway in 2001. One aim was to restrict patients from "shopping" doctors and prescription drugs. MATERIAL AND METHODS: Data from The Norwegian Prescription Database (NorPD) for 2004 were used to assess the following: number of physicians each patient had used to obtain prescriptions for a drug, the relationship between number of physicians used and amount of drugs prescribed, and concomitant prescription of opioids and benzodiazepines during the same period to the same patients. The drugs studied were the prescription drugs with potential for abuse; diazepam, carisoprodol and codeine combinations. The information retrieved was compared to that for the supposedly non-addictive drugs esomeprazole, metformine and salbutamol. RESULTS: More than 96 % (96.3 - 98.9) of the patients used one or two physicians for prescriptions of the same drug, and fewer than 0.5 % (0.01 - 0.26) used five or more physicians. However, the proportion of patients who used five or more physicians for the addictive drugs was 9.5 times higher than that for a comparable proportion of patients using the non-addictive drugs. The amount of dispensed drug increased considerably more with the number of physicians used for addictive drugs than for non-addictive drugs, as did the amount of concomitantly dispensed opioids and benzodiazepines. INTERPRETATION: The occurrence of prescription drug shopping in the Norwegian population was limited. Among the few patients who used many physicians for prescription of drugs, the phenomenon was much more frequent for users of addictive drugs than for users of the non-addictive drugs. The study suggests that prescription drug shopping may be an indicator of a drug abuse potential. PMID- 19282888 TI - [Radiation protection of childrens' chest during high resolution CT of the lungs]. AB - BACKGROUND: The Norwegian Radiation Protection Authority recommends bismuth to protect radiosensitive surface organs during CT imaging. A study based on the use of phantoms was therefore initiated to investigate the usefulness of this protection on the breasts of young girls undergoing high-resolution computed tomography (HRCT) of the chest. MATERIAL AND METHODS: Shields covered with one, two or three layers of bismuth were placed on a homogenous phantom. CT numbers (pixel values in the images which describe the density of the tissue imaged), noise and artifacts were evaluated from the images of the phantom. Dosimeters were used for dose measurements in an anthropomorphic phantom of a child. RESULTS: Areas close to the shield in the images of the phantom had CT numbers up to 33 % higher with two and three layers of bismuth than in the images without shielding. With one layer the CT numbers were up to 8 % higher. Noise in the same area was up to 56 % higher with two and three layers of bismuth than without, and up to 14 % higher with one layer. The surface dose in the breast region was reduced with 30 % with one layer of bismuth, 45 % with two and 75 % with three layers of bismuth. INTERPRETATION: Shielding with one layer of bismuth seems to have little effect on the CT images. The Department of pediatric radiology at Ullevaal University Hospital now uses bismuth protection with one layer as a routine during HRCT of girls up to 16 years of age. PMID- 19282889 TI - [Paraneoplastic neurological syndromes]. AB - BACKGROUND: Paraneoplastic neurological syndromes constitute a heterogenous group of diseases, which often cause severe neurological symptoms. Extensive research during the last 20 years has led to identification of new antigenic targets and inclusion of additional clinical conditions into this group of syndromes. MATERIAL AND METHODS: The article is based on a non-systematic search of PubMed and the author's own experience with antibody analysis, treatment of patients and research within the field. RESULTS AND INTERPRETATION: Paraneoplastic neurological syndromes affect less than 1 % of all patients with cancer. The etiology is probably autoimmune, and involves immune responses (cellular and humoral) against antigens shared by tumor cells and normal neurons. Many, but not all patients harbour onconeural antibodies (in serum and spinal fluid), which are highly useful diagnostic markers for a paraneoplastic etiology. The cornerstone of therapy is treatment of the underlying tumor, but additional immunosuppressive therapy is often administered. PMID- 19282890 TI - [Use of the dataset "The Visible Human Project" in research]. AB - The Visible Human Project has been controversial since the National Library of Medicine launched it in 1996. The project uses real bodies to create 3 dimensional virtual bodies. The first body to be photographed belonged to an executed criminal. The Medical Ethics Council of The Norwegian Medical Association has assessed whether it is ethically acceptable for an ongoing research project at the University of Oslo to use data from The Visible Human Project. PMID- 19282891 TI - [Transdermal therapeutic systems for drug administration]. AB - Transdermal therapeutic systems, or transdermal patches, facilitate controlled release of active ingredients through the skin and into the systemic circulation. Drugs administered through such systems escape first pass metabolism and steady state is maintained similar to a continuous intravenous infusion for up to several days. Few pharmacologically active substances can currently be administered through transdermal patches and production is technically demanding. Research is ongoing to improve the systems and expand the indications. This article gives a brief overview over principles behind various transdermal patch types, as well as the advantages and disadvantages of transdermal therapeutic systems. PMID- 19282892 TI - [Is gender perspective on health of the elderly necessary?]. PMID- 19282894 TI - [Deficient documentation in the guidelines for antibiotics use]. PMID- 19282895 TI - [Noscapine and warfarin--a potentially dangerous interaction]. PMID- 19282896 TI - [What is "professionally defendable"?]. PMID- 19282897 TI - [Blood specimen sending by pneumatic tube mail functions well!]. PMID- 19282899 TI - [Important witnesses in Gaza]. PMID- 19282901 TI - [Why do most of the Nobel Prizes go to USA?]. PMID- 19282903 TI - [High maternal mortality in the world's poorest countries]. PMID- 19282904 TI - [Children are killed by poisoned drugs in Nigeria]. PMID- 19282906 TI - Optimized shaping of isolated pulses in Brillouin fiber slow-light systems. AB - The effect of a proper shaping of the temporal envelope of isolated pulses in slow-light systems based on stimulated Brillouin scattering in optical fibers is studied and experimentally demonstrated. The pulse shape can be optimized to lead to a substantial enhancement of the delaying effect. The spectrum of the optical pulses is engineered so that the spectral width of the pulse is minimized while preserving the pulse duration, making possible to match at best the Brillouin spectrum. Exponentially shaped pulses show the minimal FWHM spectral width and experience the largest time delay when compared to Gaussian or rectangular pulses. PMID- 19282907 TI - Generation of high-energy sub-20 fs pulses tunable in the 250-310 nm region by frequency doubling of a high-power noncollinear optical parametric amplifier. AB - We report on the generation of powerful sub-20 fs deep UV pulses with 10 microJ level energy and broadly tunable in the 250-310 nm range. These pulses are produced by frequency doubling a high-power noncollinear optical parametric amplifier and compressed by a pair of MgF2 prisms to an almost transform-limited duration. Our results provide a power scaling by an order of magnitude with respect to previous works. PMID- 19282908 TI - Efficient upconversion of subterahertz radiation in a high-Q whispering gallery resonator. AB - We demonstrate efficient upconversion of subterahertz radiation into the optical domain in a high-Q whispering gallery mode resonator with quadratic optical nonlinearity. The 5x10(-3) power conversion efficiency of a cw 100 GHz signal is achieved with only 16 mW of optical pump. PMID- 19282909 TI - Amplification of a radially polarized laser beam using an Yb-doped double-clad fiber. AB - Amplification of a radially polarized laser beam is demonstrated by use of an Yb doped double-clad fiber whose core cutoff wavelength for the LP11 mode is larger than the wavelength of the input beam. The beam power is amplified while maintaining the intensity and polarization distributions. The amplified spontaneous emission becomes negligible with increasing input power. The measured slope efficiency is 48%, and the output power is 1.1 W. PMID- 19282910 TI - Direct determination of the generalized Stokes parameters from the usual Stokes parameters. AB - We report an experimental method to determine the generalized Stokes parameters for a pair of points in the cross section of an electromagnetic beam, e.g., an expanded laser beam, with the help of a Young's interferometer and a set of polarizers and quarter-wave plates. The method is investigated theoretically using the electromagnetic spectral interference law. The generalized Stokes parameters, owing to their two-point nature, determine the behavior of the single point polarization properties of the electromagnetic beam at a field point. The present method offers a unique means to determine the two-point parameters (correlation functions) by measuring the usual Stokes parameters (intensities) and the contrast parameters (visibilities) of the beam. The method might be applicable to determine the polarization dependent changes in various optical measurements. PMID- 19282911 TI - Realization of an evanescent Bessel beam via surface plasmon interference excited by a radially polarized beam. AB - We report the experimental confirmation of the evanescent Bessel beam generation via surface plamson resonance excitation with a radially polarized beam. The interference of surface plasmon waves excited by a radially polarized beam creates an evanescent Bessel beam with enhanced localized field and spot size beyond the diffraction limit. The excitation of the surface plasmon is confirmed by the observation of a narrow dark ring at the back focal plane. Two-dimensional intensity distributions at different distances from the sample surface are mapped by a collection-mode near-field scanning optical microscope to verify the nondiffracting and decaying natures of the evanescent Bessel beam. PMID- 19282912 TI - Subharmonic Fourier domain mode locking. AB - We demonstrate a subharmonically Fourier domain mode-locked wavelength-swept laser source with a substantially reduced cavity fiber length. In contrast to a standard Fourier domain mode-locked configuration, light is recirculated repetitively in the delay line with the optical bandpass filter used as switch. The laser has a fundamental optical round trip frequency of 285 kHz and can be operated at integer fractions thereof (subharmonics). Sweep ranges up to 95 nm full width centred at 1317 nm are achieved at the 1/5th subharmonic. A maximum sensitivity of 116 dB and an axial resolution of 12 microm in air are measured at an average sweep power of 12 mW. A sensitivity roll-off of 11 dB over 4 mm and 25 dB over 10 mm is observed and optical coherence tomography imaging is demonstrated. Besides the advantage of a reduced fiber length, subharmonic Fourier domain mode locking (shFDML) enables simple scaling of the sweep speed by extracting light from the delay part of the resonator. A sweep rate of 570 kHz is achieved. Characteristic features of shFDML operation, such as power leakage during fly-back and cw breakthrough, are investigated. PMID- 19282913 TI - Broadband omnidirectional antireflection coatings optimized by genetic algorithm. AB - An optimized graded-refractive-index (GRIN) antireflection (AR) coating with broadband and omnidirectional characteristics--as desired for solar cell applications--designed by a genetic algorithm is presented. The optimized three layer GRIN AR coating consists of a dense TiO2 and two nanoporous SiO2 layers fabricated using oblique-angle deposition. The normal incidence reflectance of the three-layer GRIN AR coating averaged between 400 and 700 nm is 3.9%, which is 37% lower than that of a conventional single-layer Si3N4 coating. Furthermore, measured reflection over the 410-740 nm range and wide incident angles 40 degrees -80 degrees is reduced by 73% in comparison with the single-layer Si3N4 coating, clearly showing enhanced omnidirectionality and broadband characteristics of the optimized three-layer GRIN AR coating. PMID- 19282914 TI - Three-dimensional recognition of photon-starved events using computational integral imaging and statistical sampling. AB - We present a statistical approach to recognize three-dimensional (3D) objects with a small number of photons captured by using integral imaging (II). For 3D recognition of the events, the photon-limited elemental image set of a 3D object is obtained using the II technique. A computational geometrical ray propagation algorithm and the parametric maximum likelihood estimator are applied to the photon-limited elemental image set to reconstruct the irradiance of the original 3D scene voxels. The sampling distributions for the statistical parameters of the reconstructed image are determined. Finally, hypothesis testing for the equality of the statistical parameters between reference and input data sets is performed for statistical classification of populations on the basis of sampling distribution information. It is shown that large data sets of photon-limited 3D images can be converted into sampling distributions with their own statistical parameters, resulting in a substantial data dimensionality reduction for processing. PMID- 19282915 TI - Double-phase conjugate mirror in nominally undoped Sn2P2S6. AB - Coherent oscillation is achieved in the geometry of a double-phase conjugate mirror in nominally undoped Sn2P2S6 with 647 nm radiation of a Kr+ laser. The specific temporal dynamics of oscillation with modulated intensity and periodic 0 pi-0-pi variations of the oscillation wave phase is similar to that observed earlier using a semilinear oscillator geometry. The described oscillator ensures submillisecond delay time in the appearance of the phase conjugate wave. PMID- 19282916 TI - Spatial solitons in liquid-crystal light valves. AB - Liquid-crystal light valves can control the orientation of a nematic layer under the independent or combined action of applied voltage and impinging light intensity; hence, they offer a unique environment for the propagation of spatial optical solitons or nematicons. We demonstrate nematicon excitation, propagation, and steering in photoconductive light valves. PMID- 19282917 TI - Microjoule pulse energy from a chirped-pulse Ti:sapphire oscillator with cavity dumping. AB - We demonstrate a chirped-pulse Ti:sapphire laser oscillator with both Kerr-lens and semiconductor- saturable-absorber-mirror-assisted mode locking generating 1.1 microJ pulses at 1 MHz pulse repetition rate. The pulses are coupled out of the laser cavity by means of an acousto-optical cavity dumper, have a spectral width that supports a Fourier limit of 74 fs, and currently have a chirped-pulse duration of 5 ps. After compressing the pulses, this laser will be an ideal tool for efficient high-harmonic generation directly from a laser oscillator. PMID- 19282918 TI - Photonic-assisted microwave frequency measurement with higher resolution and tunable range. AB - A photonic-assisted approach to microwave frequency measurement is proposed based on frequency-to-power mapping with the help of the so-called amplitude comparison function. The key component is a dual-output Mach-Zehnder modulator (MZM) working at chirped modulation. The proposed scheme is characterized as having simplicity, higher resolution, and tunable measurement range. Owing to experimental constraint, an equivalent experiment has been carried out using a common single output MZM at different bias points to prove the concept. PMID- 19282919 TI - High-efficiency 355 nm generation in barium aluminum borate diflouride (BaAlBO3F2). AB - We demonstrate high-efficiency ultraviolet (UV) laser generation in a nonlinear optical crystal BaAlBO3F2 (BABF); 355 nm UV light was generated by using a type I BABF crystal as a sum-frequency mixing of the fundamental light and the second harmonic from a picosecond Nd:YAG laser operated at a repetition rate of 10 Hz with a pulse duration of 20 ps. An output of 0.635 mJ for the third harmonic was obtained. The conversion efficiency was 26.4%. As far as we know, it was the first time that third-harmonic generation in BABF was achieved. Obviously, the BABF crystal is a promising UV nonlinear optical material because of its relatively large nonlinear coefficient and nonhygroscopicity. PMID- 19282920 TI - High-performance sensor based on surface plasmon resonance with chalcogenide prism and aluminum for detection in infrared. AB - A high-accuracy aluminum-based surface plasmon resonance (SPR) chalcogenide sensor is proposed for IR. The structure is based on widely used 2S2G chalcogenide glass with aluminum as the SPR active metal. The angular interrogation method has been used to study the performance of the sensor in terms of intrinsic sensitivity (IS) that includes the width and shifts of the SPR curve for a given refractive index of sensing layer. The IS of Al-based chalcogenide glass sensor is almost 400% more as compared with an Au-based one, which is the most widely used SPR active metal. The oxidation problem of an Al based SPR sensor has been addressed. PMID- 19282921 TI - Flat-top pulse generation based on a fiber Bragg grating in transmission. AB - We propose and analyze a flat-top pulse generator based on a fiber Bragg grating (FBG) in transmission. As is shown in the examples, a uniform period FBG properly designed can exhibit a spectral response in transmission close to sinc function (in amplitude and phase) in a certain bandwidth, because of the logarithm Hilbert transform relations, which can be used to reshape a Gaussian-like input pulse into a flat-top pulse. PMID- 19282922 TI - Lambda/2 fringe-spacing interferometer. AB - The precision of interferometry is directly linked to the fringe spacing of the recorded interferogram. Whereas all interferometric devices show a fringe spacing equal to a wavelength of the laser light we present a novel scheme of a two-beam interferometer exhibiting a fringe spacing reduced by a factor of 2; the direct detection of the beat signal is replaced with the monitoring of the fluorescence of a twofold degenerate atomic system resonant with the laser. The lambda/2 fringe spacing in the fluorescence signal is demonstrated with a hot sodium vapor excited by a broadband laser tuned to the D1 line. In the saturation regime, the dark fringes are expected to be extremely narrow, leading to the possibility of nanoscale displacement measurements or atom localization. PMID- 19282923 TI - Integrated Raman spectroscopy and trimodal wide-field imaging techniques for real time in vivo tissue Raman measurements at endoscopy. AB - We report an integrated Raman spectroscopy and trimodal (white-light reflectance, autofluorescence, and narrow-band) imaging techniques for real-time in vivo tissue Raman measurements at endoscopy. A special 1.8 mm endoscopic Raman probe with filtering modules is developed, permitting effective elimination of interference of fluorescence background and silica Raman in fibers while maximizing tissue Raman collections. We demonstrate that high-quality in vivo Raman spectra of upper gastrointestinal tract can be acquired within 1 s or subseconds under the guidance of wide-field endoscopic imaging modalities, greatly facilitating the adoption of Raman spectroscopy into clinical research and practice during routine endoscopic inspections. PMID- 19282924 TI - All-optical subdiffraction multilevel data encoding onto azo-polymeric thin films. AB - By exploiting photoinduced reorientation in azo-polymer thin films, we demonstrate all-optical polarization-encoded information storage with a scanning near-field optical microscope. In the writing routine, five-level bits are created by associating different bit values to different birefringence directions, induced in the polymer after illumination with linearly polarized light. The reading routine is then performed by implementing polarization modulation techniques on the same near-field microscope in order to measure the encoded birefringence direction. PMID- 19282925 TI - Hyperspectral imaging using a microelectrical-mechanical-systems-based in-plane vibratory grating scanner with a single photodetector. AB - We present a single-photodetector-based hyperspectral imaging system that utilizes a microelectrical-mechanical-systems-driven diffraction grating for fast spatial scanning and two synchronized steering mirrors for slow spectral scanning. This configuration allows high-speed scanning without degradation in optical performance resulting from dynamic non-rigid-body deformation of the platform. The proposed operational principle is demonstrated with a prototype device developed using silicon microfabrication technology. The proposed spectral imaging system has the potential to achieve low cost, small form factor, and high speed operation. PMID- 19282926 TI - Dual-interference-channel quantitative-phase microscopy of live cell dynamics. AB - We introduce and experimentally demonstrate a fast and accurate method for quantitative imaging of the dynamics of live biological cells. Using a dual channel interferometric setup, two phase-shifted interferograms of nearly transparent biological samples are acquired in a single digital camera exposure and digitally processed into the phase profile of the sample. Since two interferograms of the same sample are acquired simultaneously, most of the common phase noise is eliminated, enabling the visualization of millisecond-scale dynamic biological phenomena with subnanometer optical path length temporal stability. PMID- 19282927 TI - Two-dimensional solitons in nonlinear lattices. AB - We address the existence and stability of two-dimensional solitons in optical or matter-wave media, which are supported by purely nonlinear lattices in the form of a periodic array of cylinders with self-focusing nonlinearity, embedded into a linear material. We show that such lattices can stabilize two-dimensional solitons against collapse. We also found that stable multipoles and vortex solitons are also supported by nonlinear lattices, provided that the nonlinearity exhibits saturation. PMID- 19282928 TI - Comparing coherent and spontaneous Raman scattering under biological imaging conditions. AB - We compare imaging based on coherent and spontaneous Raman scattering (SpRS) under conditions relevant for biological imaging. Using a broadband laser source, we perform spectral domain imaging of polystyrene beads using coherent Stokes Raman scattering and SpRS and find comparable signal levels. Short interaction lengths, low molecule number, and low incident power all reduce the advantages available with coherent Raman methods. We present calculations to support our measurements. PMID- 19282929 TI - Atomic coherence swing in a double-Lambda-type system using ultraslow light. AB - Using an ultraslow light-based double-Lambda-type configuration we demonstrate atomic coherence swing between two different frequencies in a nondegenerate four wave mixing scheme. A short control pulse applied to the middle of an ultraslow light envelope results in coherence conversion between two ground states, causing both coherence depletion and recovery, resulting in a temporal hole and amplification in the ultraslow light envelope. This outcome holds potential for temporally and spatially localized coherence control of ultraslow light in such applications as a selective photon trap. PMID- 19282930 TI - Fundamental limit for two-dimensional passive devices. AB - There is an upper limit to the number of electromagnetic communication channels in two-dimensional linear passive components that depends only on the geometrical dimensions but is independent of the permittivity function, the actual external shape, or the internal design. The limit applies to both weakly and strongly scattering waves. When the permittivity contrast is low, a tighter limit exists that includes only multiple scattering waves. A detailed analysis helps compare these two limits and leads to insights that apply to devices, such as photonic crystals and microresonators, as well as lossless metamaterials, superlenses, and cloaking devices. As an example, we establish a rigorous scaling relation for the upper bound of the number of demultiplexing channels in superprisms. PMID- 19282931 TI - Power scaling of Nd:YVO4 and Nd:GdVO4 disk lasers using synthetic diamond as a heat spreader. AB - A newly developed low-birefringence synthetic diamond is shown to be an effective intracavity heat spreader in Nd:YVO4 and Nd:GdVO4 disk lasers. A cw output power of 25.7 W from only one double pass of the pump is reported. The diamond heat spreader is shown to increase the pump power density at which fracture occurs. PMID- 19282932 TI - Chirp-free optical modulation using a silicon push-pull coupling microring. AB - We propose a silicon ring-based optical modulation method to perform chirp-free optical modulations. In this scheme, we locate the light to be modulated at the resonance of the ring and tune the coupling coefficient between the ring and the straight waveguide by using a push-pull coupling structure. The chirp-free phase modulation can be achieved by varying the coupling coefficient in a large range, which can modify the coupling condition of the ring such that the input light experiences an abrupt phase shift of pi at the output. If the coupling coefficient is adjusted in a small range such that the coupling condition of the ring is kept unchanged, only the intensity of the light will be modulated. This leads to chirp-free intensity modulation. Our simulations performed at 10 Gbits/s confirm the feasibility of the proposal. PMID- 19282933 TI - Complex periodic micro/nanostructures on 6H-SiC crystal induced by the interference of three femtosecond laser beams. AB - We reported three types of complex micro/nanostructures on 6H-SiC crystal induced by the interferences of three femtosecond laser beams by arranging three types of laser polarization combinations. The micro/nanostructures are composed of two parts: two-dimensional long-periodic micropatterns determined by the interferential intensity pattern and short-periodic nanopatterns determined by the interferential polarization pattern. Theoretical calculation indicates that the different polarization combinations will lead to a distinct complex interferential polarization pattern and intensity pattern, and they accord well with the experimental results. PMID- 19282934 TI - All-optical switching application of germano-silicate optical fiber incorporated with Ag nanocrystals. AB - Ag-nanocrystal-incorporated germano-silicate optical fiber with high resonant nonlinearity was fabricated by using modified chemical vapor deposition and solution doping techniques. An all-optical signal switching application based on cross-phase modulation was demonstrated in the cascaded long-period fiber gratings. Pumped with 499 nm argon laser, all-optical signal gating of the pi phase shift was achieved with low pump intensities of less than 7.64 GW/cm2. PMID- 19282935 TI - Photoluminescent and decay behaviors of Mn2+ and Ce3+ coactivated MgSiN2 phosphors for use in light-emitting-diode applications. AB - We examined the photoluminescent behaviors of MgSiN2:Mn2+ and MgSiN2:Ce3+,Mn2+ phosphors for use in white-light-emitting diodes. The red emission from MgSiN2:Mn2+ phosphors consisted of two Gaussian components, P1 from a single Mn2+ ion and P2 from either Mn2+ pairs or clusters. Decay analysis based on the Yokota and Tanimoto equation identified long decay for P1 and fast decay for P2. Most importantly, Ce3+ codoping enhanced Mn2+ emission intensity; in particular, emission at 460 nm excitations was promoted by the Ce3+ codoping. PMID- 19282936 TI - Observation of two-dimensional defect surface solitons. AB - We report on the experimental observation of two-dimensional solitons located in defect channels at the surface of a hexagonal waveguide array. The threshold power for the excitation of solitons existing owing to total internal reflection grows with decrease of the refractive index in negative defects and vanishes for sufficiently strong positive defects. Negative defects can also support linear surface modes existing owing to Bragg-type reflections. PMID- 19282937 TI - Measurement of Mount Etna plume by CO2-laser-based lidar. AB - The CO2 laser-based agile tuner lidar for atmospheric sensing has been used to profile the volcanic plume of Mount Etna during its most recent eruption. Owing to the transmitted wavelength, this system is practically insensitive to air molecules while it detects aerosol loads, and thus the path attenuation of the laser beam is strongly affected by volcanic particulate. Vertical profiles of extinction coefficient were retrieved up to an altitude above ground level of 5000 m. The observed extinction coefficient ranges from 10(-5) to 5x10(-4) m(-1). The lidar was able to accurately track the spatiotemporal evolution of the volcanic plume thanks to a spatial resolution of 15 m and a temporal resolution of 1 min. PMID- 19282938 TI - Variable phase retarder made of a dielectric elastomer actuator. AB - We present a polymeric optical phase retarder that is electrically tunable by a dielectric elastomer actuator. The soft material device affords a large tuning range (14pi at lambda=488 nm) combined with high accuracy in optical path length and low drift rate (8.3 nm/min). Furthermore, the phase retarder is not sensitive to polarization, introduces a wavefront distortion141 kW/cm2). We show the dynamics for periodic phase modulation and demonstrate a simple drive technique for fast phase stepping. The polymer based device is inexpensive, easy to fabricate, and its design can be adapted to specific applications. PMID- 19282939 TI - Simultaneous measurement of in-plane and out-of-plane displacement fields in scattering media using phase-contrast spectral optical coherence tomography. AB - The use of phase-contrast spectral optical coherence tomography to measure two orthogonal displacement components on a slice within a scattering medium is demonstrated. This is achieved by combining sequential oblique illumination of the object and recording two interferograms before plus two after the deformation. The proposed technique is illustrated with results from a sample undergoing simple shear. Depth-resolved out-of-plane and in-plane sensitivities of 0.14 and 4.2 microm per fringe are demonstrated up to a depth of 400 microm in a water-based polymer. PMID- 19282940 TI - Fresnel zone antenna for dual-band detection at millimeter and infrared wavelengths. AB - In this work the concept of a Fresnel zone antenna for dual-band detection in the IR and millimeter wave region is presented. The design is based on a Fresnel zone plate lens in the IR that is transformed to serve as a millimeter-wave antenna. Two different designs are presented, a circular-zone design that gives a high diffractive efficiency in the IR and a square-zone design that gives a higher response in the millimeter band but a lower focusing efficiency in the IR. Both designs have an operation bandwidth with the same low frequency limit of 400 GHz (750 microm), which can be tailored by changing the number of Fresnel zones, and a high frequency limit of 4.5 THz (65 microm) for the circular-zone design and 5 THz (59 microm) for the square-zone design. PMID- 19282941 TI - All-optical frequency conversion using nonlinear dynamics of semiconductor lasers. AB - When a semiconductor laser is subject to optical injection, it can enter the period-one dynamics through Hopf bifurcation. Under such nonlinear dynamics, equally and oppositely frequency-shifted optical signals from the injection emerge and are utilized for frequency conversion. Only a typical semiconductor laser is required as the conversion unit, where no pump or probe laser is necessary. The frequency shift can be continuously tuned by controlling the level or frequency of the injection. A bit-error ratio down to 10(-12) is observed with no or a slight power penalty for amplitude, frequency, and phase modulation at 2.5 Gbits/s, suggesting modulation format transparency of the system. Frequency down-, no-, and upconversion can be simultaneously achieved and individually selected, increasing the flexibility and reconfigurability of the system. PMID- 19282942 TI - Complete elimination of self-pulsations in dual-clad ytterbium-doped fiber lasers at all pumping levels. AB - We have demonstrated suppression and elimination of self-pulsing in a watt-level, dual-clad, ytterbium-doped fiber laser. The addition of a long section of passive fiber in the laser cavity makes the gain recovery faster than the self-pulsation dynamics, allowing only stable cw lasing. This scheme provides a simple and practical method for eliminating self-pulsations in fiber lasers at all pumping levels. PMID- 19282943 TI - Broadband multilayer-coated normal incidence blazed grating with approximately 10% diffraction efficiency through the 13-16 nm wavelength region. AB - Diffraction gratings used in extreme UV are typically coated with periodic multilayer thin films. These coatings have a small bandwidth, thus leading to a narrow usable spectral region of multilayer gratings. Well-designed aperiodic multilayer coatings could provide high reflectivity over a much broader wavelength region, so they could broaden the usable spectral region of multilayer gratings. We designed and deposited an aperiodic Mo/Si multilayer coating onto a blazed grating substrate. At an incidence angle of 10 degrees, the -2nd-order diffraction efficiency of the multilayer grating is approximately 10% through the wavelength range of 13-16 nm. PMID- 19282944 TI - Motion control of low-index microspheres in liquid based on optical repulsive force of a focused beam array. AB - In the optical tweezers technique a microdielectric object with a higher refractive index than a surrounding liquid is trapped near the focal point of a laser beam, whereas an object with a lower refractive index is subjected to a repulsive force from the focused beam. We demonstrate that the optical repulsive forces are generated with a focused-beam array dynamically formed by a computer generated hologram displayed on a liquid-crystal spatial light modulator. We also demonstrate that the optical repulsive forces perform a size selection and flow control of hollow glass microspheres. PMID- 19282945 TI - Broadband squeezing of quantum noise in a Michelson interferometer with Twin Signal-Recycling. AB - Twin-Signal-Recycling (TSR) builds on the resonance doublet of two optically coupled cavities and efficiently enhances the sensitivity of an interferometer at a dedicated signal frequency. We report on what we believe to be the first experimental realization of a TSR Michelson interferometer and also its broadband enhancement by squeezed light injection. The complete setup was stably locked, and a broadband quantum noise reduction of the interferometers shot noise by a factor of up to 4 dB was demonstrated. The system was characterized by measuring its quantum noise spectra for several tunings of the TSR cavities. We found good agreement between the experimental results and numerical simulations. PMID- 19282946 TI - Spectral modulation of femtosecond laser pulse induced by molecular alignment revivals. AB - We demonstrate experimentally and numerically that the quantum wake of molecular alignment induced by impulsive rotational Raman excitations with femtosecond pump pulses produces observable phase modulations on femtosecond probe pulses. This leads to a spectral red- or blueshift of the probe pulses when they are properly delayed around the rising or falling edge of the half-revival time of the molecular alignment, respectively. PMID- 19282947 TI - All-optical differentiator and high-speed pulse generation based on cross polarization modulation in a semiconductor optical amplifier. AB - We propose an all-optical intensity differentiation scheme based on cross polarization modulation (XPolM) in a semiconductor optical amplifier (SOA) while demonstrating the absolute value of differential signal that can be obtained by the SOA-based XPolM of two parts with relative delay from the input signal and well extracted by the polarization filter. The differentiation errors and eye diagrams versus sampling time Delta are investigated for data rate at 12.5 Gbits/s, and the minimal error approximately 0.06 is achieved at Delta=0. Owing to a much faster polarization response, our scheme bears great potential for all optical signal processing over 100 Gbits/s. By application of the differentiator, we further obtain the 20 GHz short pulse train with a pulse width of approximately 10 ps. PMID- 19282948 TI - In-fiber all-optical fractional differentiator. AB - We demonstrate that an asymmetrical pi phase-shifted fiber Bragg grating operated in reflection can provide the required spectral response for implementing an all optical fractional differentiator. There are different (but equivalent) ways to design it, e.g., by using different gratings lengths and keeping the same index modulation depth at both sides of the pi phase shift, or vice versa. Analytical expressions were found relating the fractional differentiator order with the grating parameters. The device shows a good accuracy calculating the fractional time derivatives of the complex field of an arbitrary input optical waveform. The introduced concept is supported by numerical simulations. PMID- 19282949 TI - Fast two-dimensional standing-wave total-internal-reflection fluorescence microscopy using acousto-optic deflectors. AB - We report a scheme for 2D standing-wave total-internal-reflection fluorescence microscopy. Standing-wave patterns are generated by two interfering beams coupled through the objective lens. Period, angular orientation, and phase of standing waves are controlled entirely by acousto-optic deflectors. The lateral resolution improvement of 100 nm is combined with an axial selectivity of <100 nm by utilizing an evanescent standing-wave pattern. This technique can provide real time imaging of subresolution structures in live biological specimens near a glass-water interface. PMID- 19282950 TI - Thickness dependence of surface plasmon polariton dispersion in transparent conducting oxide films at 1.55 microm. AB - We experimentally demonstrate propagation of surface plasmon polaritons in the near-IR window lambda (1.45 microm,1.59 microm) at the interface of indium-tin oxide films with different thicknesses deposited on glass. Dispersion of such polaritons is strongly dependent on the film thickness, putting into evidence a regime in which polaritons at both films's interfaces are coupled in surface supermodes. The experimental data are shown to be in good agreement with the analytical model for thin and absorbing conducting films. Measurements on aluminum-doped zinc oxide, characterized by a redshifted plasma resonance, do not show any surface plasmon polariton excitation in the same wavelength window. PMID- 19282951 TI - Femtosecond soliton source with fast and broad spectral tunability. AB - We present a complete set of measurements and numerical simulations of a femtosecond soliton source with fast and broad spectral tunability and nearly constant pulse width and average power. Solitons generated in a photonic crystal fiber, at the low-power coupling regime, can be tuned in a broad range of wavelengths, from 850 to 1200 nm using the input power as the control parameter. These solitons keep almost constant time duration (approximately 40 fs) and spectral widths (approximately 20 nm) over the entire measured spectra regardless of input power. Our numerical simulations agree well with measurements and predict a wide working wavelength range and robustness to input parameters. PMID- 19282952 TI - Excitation of gap modes in a metal particle-surface system for sub-30 nm plasmonic lithography. AB - In this Letter, a near-field optical excitation of gap modes in a metal particle surface system for patterning periodic nanostructure is proposed and numerically demonstrated using the finite-difference time-domain method. It is observed that high-density sub-30 nm periodic structures were achievable by employing an aluminium nanosphere-silver surface system. A 2D resist profile cross section using the modified cellular automata model, which was obtained through this proposed configuration, is also presented. PMID- 19282953 TI - Multiorder nonlinear diffraction in frequency doubling processes. AB - We analyze experimentally light scattering from chi(2) nonlinear gratings and observe two types of second-harmonic frequency-scattering processes. The first process is identified as Raman-Nath type nonlinear diffraction that is explained by applying only transverse phase-matching conditions. The angular position of this type of diffraction is defined by the ratio of the second-harmonic wavelength and the grating period. In contrast, the second type of nonlinear scattering process is explained by the longitudinal phase matching only, being insensitive to the nonlinear grating period. PMID- 19282954 TI - Powerful wavelength-tunable ultrashort solitons in a solid-core photonic-crystal fiber. AB - A solid-core photonic-crystal fiber (PCF) with an effective mode area of 20 microm2 is used to demonstrate the generation of sub-100-kW, 30-70 fs wavelength tunable solitons within a wavelength range of 1300-1800 nm at a repetition rate of 18 MHz. An energy of 2.9 nJ per pulse is achieved for a 35 fs soliton PCF output centered at 1770 nm. Our numerical analysis supports experimental findings and suggests that frequency-shifted solitons in solid-core PCFs can be scaled up to a submegawatt level of peak powers. PMID- 19282955 TI - Coherent ultrafast pulse synthesis between an optical parametric oscillator and a laser. AB - We have demonstrated coherent pulse synthesis between the carrier-envelope, phase locked, second-harmonic pulses from a synchronously pumped femtosecond optical parametric oscillator and those from its self-mode-locked Ti:sapphire pump laser. By using a single nonlinear crystal for parametric and second-harmonic generation, we maximized the common-mode rejection of environmental noise, obtaining a temporal overlap between the pulses with a precision of 30 as (1% of the optical period) in an observation time of 20 ms. Mutual coherence between the two parent pulses was verified optically by spectral interferometry, and synthesis was tested by measuring the autocorrelation of the combined pulses, with and without carrier-envelope phase locking. PMID- 19282956 TI - Sizing of particles smaller than 5 microm in digital holographic microscopy. AB - The holographic microscopy technique is a strong contender for dynamic three dimensional (3D) measurement of small particles (typically smaller than 5 microm) in microchannels. However, there is a big challenge to accurately measure the size of such small particles. The traditional hologram reconstruction method was numerically investigated. It is found that the error level, especially for the size measurement, is higher than expected, even in an ideal situation without consideration of noise. An alternative way based on Lorenz-Mie (LM) calculations was then presented. The intensity distribution of the fringe pattern on the particle hologram is directly used and compared with the ones calculated using an LM-based program. Various cases for particle sizes from 0.5 to 5 microm and recording distances from 5 to 500 microm are tested. The results show that the accuracy in particle sizing can be significantly improved. PMID- 19282957 TI - Hybrid transfer-matrix FDTD method for layered periodic structures. AB - A hybrid transfer-matrix finite-difference time-domain (FDTD) method is proposed for modeling the optical properties of finite-width planar periodic structures. This method can also be applied for calculation of the photonic bands in infinite photonic crystals. We describe the procedure of evaluating the transfer-matrix elements by a special numerical FDTD simulation. The accuracy of the new method is tested by comparing computed transmission spectra of a 32-layered photonic crystal composed of spherical or ellipsoidal scatterers with the results of direct FDTD and layer-multiple-scattering calculations. PMID- 19282958 TI - Timing the emergence of resistance to anti-HIV drugs with large genetic barriers. AB - New antiretroviral drugs that offer large genetic barriers to resistance, such as the recently approved inhibitors of HIV-1 protease, tipranavir and darunavir, present promising weapons to avert the failure of current therapies for HIV infection. Optimal treatment strategies with the new drugs, however, are yet to be established. A key limitation is the poor understanding of the process by which HIV surmounts large genetic barriers to resistance. Extant models of HIV dynamics are predicated on the predominance of deterministic forces underlying the emergence of resistant genomes. In contrast, stochastic forces may dominate, especially when the genetic barrier is large, and delay the emergence of resistant genomes. We develop a mathematical model of HIV dynamics under the influence of an antiretroviral drug to predict the waiting time for the emergence of genomes that carry the requisite mutations to overcome the genetic barrier of the drug. We apply our model to describe the development of resistance to tipranavir in in vitro serial passage experiments. Model predictions of the times of emergence of different mutant genomes with increasing resistance to tipranavir are in quantitative agreement with experiments, indicating that our model captures the dynamics of the development of resistance to antiretroviral drugs accurately. Further, model predictions provide insights into the influence of underlying evolutionary processes such as recombination on the development of resistance, and suggest guidelines for drug design: drugs that offer large genetic barriers to resistance with resistance sites tightly localized on the viral genome and exhibiting positive epistatic interactions maximally inhibit the emergence of resistant genomes. PMID- 19282959 TI - Mechanism of action of cyclophilin a explored by metadynamics simulations. AB - Trans/cis prolyl isomerisation is involved in several biological processes, including the development of numerous diseases. In the HIV-1 capsid protein (CA), such a process takes place in the uncoating and recruitment of the virion and is catalyzed by cyclophilin A (CypA). Here, we use metadynamics simulations to investigate the isomerization of CA's model substrate HAGPIA in water and in its target protein CypA. Our results allow us to propose a novel mechanistic hypothesis, which is finally consistent with all of the available molecular biology data. PMID- 19282960 TI - Mechanical network in titin immunoglobulin from force distribution analysis. AB - The role of mechanical force in cellular processes is increasingly revealed by single molecule experiments and simulations of force-induced transitions in proteins. How the applied force propagates within proteins determines their mechanical behavior yet remains largely unknown. We present a new method based on molecular dynamics simulations to disclose the distribution of strain in protein structures, here for the newly determined high-resolution crystal structure of I27, a titin immunoglobulin (IG) domain. We obtain a sparse, spatially connected, and highly anisotropic mechanical network. This allows us to detect load-bearing motifs composed of interstrand hydrogen bonds and hydrophobic core interactions, including parts distal to the site to which force was applied. The role of the force distribution pattern for mechanical stability is tested by in silico unfolding of I27 mutants. We then compare the observed force pattern to the sparse network of coevolved residues found in this family. We find a remarkable overlap, suggesting the force distribution to reflect constraints for the evolutionary design of mechanical resistance in the IG family. The force distribution analysis provides a molecular interpretation of coevolution and opens the road to the study of the mechanism of signal propagation in proteins in general. PMID- 19282961 TI - Cortical factor feedback model for cellular locomotion and cytofission. AB - Eukaryotic cells can move spontaneously without being guided by external cues. For such spontaneous movements, a variety of different modes have been observed, including the amoeboid-like locomotion with protrusion of multiple pseudopods, the keratocyte-like locomotion with a widely spread lamellipodium, cell division with two daughter cells crawling in opposite directions, and fragmentations of a cell to multiple pieces. Mutagenesis studies have revealed that cells exhibit these modes depending on which genes are deficient, suggesting that seemingly different modes are the manifestation of a common mechanism to regulate cell motion. In this paper, we propose a hypothesis that the positive feedback mechanism working through the inhomogeneous distribution of regulatory proteins underlies this variety of cell locomotion and cytofission. In this hypothesis, a set of regulatory proteins, which we call cortical factors, suppress actin polymerization. These suppressing factors are diluted at the extending front and accumulated at the retracting rear of cell, which establishes a cellular polarity and enhances the cell motility, leading to the further accumulation of cortical factors at the rear. Stochastic simulation of cell movement shows that the positive feedback mechanism of cortical factors stabilizes or destabilizes modes of movement and determines the cell migration pattern. The model predicts that the pattern is selected by changing the rate of formation of the actin-filament network or the threshold to initiate the network formation. PMID- 19282962 TI - Toward a detailed description of the thermally induced dynamics of the core promoter. AB - Establishing the general and promoter-specific mechanistic features of gene transcription initiation requires improved understanding of the sequence dependent structural/dynamic features of promoter DNA. Experimental data suggest that a spontaneous dsDNA strand separation at the transcriptional start site is likely to be a requirement for transcription initiation in several promoters. Here, we use Langevin molecular dynamic simulations based on the Peyrard-Bishop Dauxois nonlinear model of DNA (PBD LMD) to analyze the strand separation (bubble) dynamics of 80-bp-long promoter DNA sequences. We derive three dynamic criteria, bubble probability, bubble lifetime, and average strand separation, to characterize bubble formation at the transcriptional start sites of eight mammalian gene promoters. We observe that the most stable dsDNA openings do not necessarily coincide with the most probable openings and the highest average strand displacement, underscoring the advantages of proper molecular dynamic simulations. The dynamic profiles of the tested mammalian promoters differ significantly in overall profile and bubble probability, but the transcriptional start site is often distinguished by large (longer than 10 bp) and long-lived transient openings in the double helix. In support of these results are our experimental transcription data demonstrating that an artificial bubble containing DNA template is transcribed bidirectionally by human RNA polymerase alone in the absence of any other transcription factors. PMID- 19282963 TI - Probabilistic interaction network of evidence algorithm and its application to complete labeling of peak lists from protein NMR spectroscopy. AB - The process of assigning a finite set of tags or labels to a collection of observations, subject to side conditions, is notable for its computational complexity. This labeling paradigm is of theoretical and practical relevance to a wide range of biological applications, including the analysis of data from DNA microarrays, metabolomics experiments, and biomolecular nuclear magnetic resonance (NMR) spectroscopy. We present a novel algorithm, called Probabilistic Interaction Network of Evidence (PINE), that achieves robust, unsupervised probabilistic labeling of data. The computational core of PINE uses estimates of evidence derived from empirical distributions of previously observed data, along with consistency measures, to drive a fictitious system M with Hamiltonian H to a quasi-stationary state that produces probabilistic label assignments for relevant subsets of the data. We demonstrate the successful application of PINE to a key task in protein NMR spectroscopy: that of converting peak lists extracted from various NMR experiments into assignments associated with probabilities for their correctness. This application, called PINE-NMR, is available from a freely accessible computer server (http://pine.nmrfam.wisc.edu). The PINE-NMR server accepts as input the sequence of the protein plus user-specified combinations of data corresponding to an extensive list of NMR experiments; it provides as output a probabilistic assignment of NMR signals (chemical shifts) to sequence-specific backbone and aliphatic side chain atoms plus a probabilistic determination of the protein secondary structure. PINE-NMR can accommodate prior information about assignments or stable isotope labeling schemes. As part of the analysis, PINE-NMR identifies, verifies, and rectifies problems related to chemical shift referencing or erroneous input data. PINE-NMR achieves robust and consistent results that have been shown to be effective in subsequent steps of NMR structure determination. PMID- 19282964 TI - GrowMatch: an automated method for reconciling in silico/in vivo growth predictions. AB - Genome-scale metabolic reconstructions are typically validated by comparing in silico growth predictions across different mutants utilizing different carbon sources with in vivo growth data. This comparison results in two types of model prediction inconsistencies; either the model predicts growth when no growth is observed in the experiment (GNG inconsistencies) or the model predicts no growth when the experiment reveals growth (NGG inconsistencies). Here we propose an optimization-based framework, GrowMatch, to automatically reconcile GNG predictions (by suppressing functionalities in the model) and NGG predictions (by adding functionalities to the model). We use GrowMatch to resolve inconsistencies between the predictions of the latest in silico Escherichia coli (iAF1260) model and the in vivo data available in the Keio collection and improved the consistency of in silico with in vivo predictions from 90.6% to 96.7%. Specifically, we were able to suggest consistency-restoring hypotheses for 56/72 GNG mutants and 13/38 NGG mutants. GrowMatch resolved 18 GNG inconsistencies by suggesting suppressions in the mutant metabolic networks. Fifteen inconsistencies were resolved by suppressing isozymes in the metabolic network, and the remaining 23 GNG mutants corresponding to blocked genes were resolved by suitably modifying the biomass equation of iAF1260. GrowMatch suggested consistency-restoring hypotheses for five NGG mutants by adding functionalities to the model whereas the remaining eight inconsistencies were resolved by pinpointing possible alternate genes that carry out the function of the deleted gene. For many cases, GrowMatch identified fairly nonintuitive model modification hypotheses that would have been difficult to pinpoint through inspection alone. In addition, GrowMatch can be used during the construction phase of new, as opposed to existing, genome scale metabolic models, leading to more expedient and accurate reconstructions. PMID- 19282965 TI - Canalization of gene expression and domain shifts in the Drosophila blastoderm by dynamical attractors. AB - The variation in the expression patterns of the gap genes in the blastoderm of the fruit fly Drosophila melanogaster reduces over time as a result of cross regulation between these genes, a fact that we have demonstrated in an accompanying article in PLoS Biology (see Manu et al., doi:10.1371/journal.pbio.1000049). This biologically essential process is an example of the phenomenon known as canalization. It has been suggested that the developmental trajectory of a wild-type organism is inherently stable, and that canalization is a manifestation of this property. Although the role of gap genes in the canalization process was established by correctly predicting the response of the system to particular perturbations, the stability of the developmental trajectory remains to be investigated. For many years, it has been speculated that stability against perturbations during development can be described by dynamical systems having attracting sets that drive reductions of volume in phase space. In this paper, we show that both the reduction in variability of gap gene expression as well as shifts in the position of posterior gap gene domains are the result of the actions of attractors in the gap gene dynamical system. Two biologically distinct dynamical regions exist in the early embryo, separated by a bifurcation at 53% egg length. In the anterior region, reduction in variation occurs because of stability induced by point attractors, while in the posterior, the stability of the developmental trajectory arises from a one-dimensional attracting manifold. This manifold also controls a previously characterized anterior shift of posterior region gap domains. Our analysis shows that the complex phenomena of canalization and pattern formation in the Drosophila blastoderm can be understood in terms of the qualitative features of the dynamical system. The result confirms the idea that attractors are important for developmental stability and shows a richer variety of dynamical attractors in developmental systems than has been previously recognized. PMID- 19282966 TI - Multifactorial regulation of a hox target gene. AB - Hox proteins play fundamental roles in controlling morphogenetic diversity along the anterior-posterior body axis of animals by regulating distinct sets of target genes. Within their rather broad expression domains, individual Hox proteins control cell diversification and pattern formation and consequently target gene expression in a highly localized manner, sometimes even only in a single cell. To achieve this high-regulatory specificity, it has been postulated that Hox proteins co-operate with other transcription factors to activate or repress their target genes in a highly context-specific manner in vivo. However, only a few of these factors have been identified. Here, we analyze the regulation of the cell death gene reaper (rpr) by the Hox protein Deformed (Dfd) and suggest that local activation of rpr expression in the anterior part of the maxillary segment is achieved through a combinatorial interaction of Dfd with at least eight functionally diverse transcriptional regulators on a minimal enhancer. It follows that context-dependent combinations of Hox proteins and other transcription factors on small, modular Hox response elements (HREs) could be responsible for the proper spatio-temporal expression of Hox targets. Thus, a large number of transcription factors are likely to be directly involved in Hox target gene regulation in vivo. PMID- 19282967 TI - Intrinsic disorder in protein interactions: insights from a comprehensive structural analysis. AB - We perform a large-scale study of intrinsically disordered regions in proteins and protein complexes using a non-redundant set of hundreds of different protein complexes. In accordance with the conventional view that folding and binding are coupled, in many of our cases the disorder-to-order transition occurs upon complex formation and can be localized to binding interfaces. Moreover, analysis of disorder in protein complexes depicts a significant fraction of intrinsically disordered regions, with up to one third of all residues being disordered. We find that the disorder in homodimers, especially in symmetrical homodimers, is significantly higher than in heterodimers and offer an explanation for this interesting phenomenon. We argue that the mechanisms of regulation of binding specificity through disordered regions in complexes can be as common as for unbound monomeric proteins. The fascinating diversity of roles of disordered regions in various biological processes and protein oligomeric forms shown in our study may be a subject of future endeavors in this area. PMID- 19282968 TI - Escherichia coli MazF leads to the simultaneous selective synthesis of both "death proteins" and "survival proteins". AB - The Escherichia coli mazEF module is one of the most thoroughly studied toxin antitoxin systems. mazF encodes a stable toxin, MazF, and mazE encodes a labile antitoxin, MazE, which prevents the lethal effect of MazF. MazF is an endoribonuclease that leads to the inhibition of protein synthesis by cleaving mRNAs at ACA sequences. Here, using 2D-gels, we show that in E. coli, although MazF induction leads to the inhibition of the synthesis of most proteins, the synthesis of an exclusive group of proteins, mostly smaller than about 20 kDa, is still permitted. We identified some of those small proteins by mass spectrometry. By deleting the genes encoding those proteins from the E. coli chromosome, we showed that they were required for the death of most of the cellular population. Under the same experimental conditions, which induce mazEF-mediated cell death, other such proteins were found to be required for the survival of a small sub population of cells. Thus, MazF appears to be a regulator that induces downstream pathways leading to death of most of the population and the continued survival of a small sub-population, which will likely become the nucleus of a new population when growth conditions become less stressful. PMID- 19282969 TI - A fine-structure map of spontaneous mitotic crossovers in the yeast Saccharomyces cerevisiae. AB - Homologous recombination is an important mechanism for the repair of DNA damage in mitotically dividing cells. Mitotic crossovers between homologues with heterozygous alleles can produce two homozygous daughter cells (loss of heterozygosity), whereas crossovers between repeated genes on non-homologous chromosomes can result in translocations. Using a genetic system that allows selection of daughter cells that contain the reciprocal products of mitotic crossing over, we mapped crossovers and gene conversion events at a resolution of about 4 kb in a 120-kb region of chromosome V of Saccharomyces cerevisiae. The gene conversion tracts associated with mitotic crossovers are much longer (averaging about 12 kb) than the conversion tracts associated with meiotic recombination and are non-randomly distributed along the chromosome. In addition, about 40% of the conversion events have patterns of marker segregation that are most simply explained as reflecting the repair of a chromosome that was broken in G1 of the cell cycle. PMID- 19282970 TI - Kaposi's sarcoma-associated herpesvirus ORF45 interacts with kinesin-2 transporting viral capsid-tegument complexes along microtubules. AB - Open reading frame (ORF) 45 of Kaposi's sarcoma-associated herpesvirus (KSHV) is a tegument protein. A genetic analysis with a null mutant suggested a possible role for this protein in the events leading to viral egress. In this study, ORF45 was found to interact with KIF3A, a kinesin-2 motor protein that transports cargoes along microtubules to cell periphery in a yeast two-hybrid screen. The association was confirmed by both co-immunoprecipitation and immunoflorescence approaches in primary effusion lymphoma cells following virus reactivation. ORF45 principally mediated the docking of entire viral capsid-tegument complexes onto the cargo-binding domain of KIF3A. Microtubules served as the major highways for transportation of these complexes as evidenced by drastically reduced viral titers upon treatment of cells with a microtubule depolymerizer, nocodazole. Confocal microscopic images further revealed close association of viral particles with microtubules. Inhibition of KIF3A-ORF45 interaction either by the use of a headless dominant negative (DN) mutant of KIF3A or through shRNA-mediated silencing of endogenous KIF3A expression noticeably decreased KSHV egress reflecting as appreciable reductions in the release of extracellular virions. Both these approaches, however, failed to impact HSV-1 egress, demonstrating the specificity of KIF3A in KSHV transportation. This study thus reports on transportation of KSHV viral complexes on microtubules by KIF3A, a kinesin motor thus far not implicated in virus transportation. All these findings shed light on the understudied but significant events in the KSHV life cycle, delineating a crucial role of a KSHV tegument protein in cellular transport of viral particles. PMID- 19282971 TI - Caspar controls resistance to Plasmodium falciparum in diverse anopheline species. AB - Immune responses mounted by the malaria vector Anopheles gambiae are largely regulated by the Toll and Imd (immune deficiency) pathways via the NF-kappaB transcription factors Rel1 and Rel2, which are controlled by the negative regulators Cactus and Caspar, respectively. Rel1- and Rel2-dependent transcription in A. gambiae has been shown to be particularly critical to the mosquito's ability to manage infection with the rodent malaria parasite Plasmodium berghei. Using RNA interference to deplete the negative regulators of these pathways, we found that Rel2 controls resistance of A. gambiae to the human malaria parasite Plasmodium falciparum, whereas Rel 1 activation reduced infection levels. The universal relevance of this defense system across Anopheles species was established by showing that caspar silencing also prevents the development of P. falciparum in the major malaria vectors of Asia and South America, A. stephensi and A. albimanus, respectively. Parallel studies suggest that while Imd pathway activation is most effective against P. falciparum, the Toll pathway is most efficient against P. berghei, highlighting a significant discrepancy between the human pathogen and its rodent model. High throughput gene expression analyses identified a plethora of genes regulated by the activation of the two Rel factors and revealed that the Toll pathway played a more diverse role in mosquito biology than the Imd pathway, which was more immunity-specific. Further analyses of key anti-Plasmodium factors suggest they may be responsible for the Imd pathway-mediated resistance phenotype. Additionally, we found that the fitness cost caused by Rel2 activation through caspar gene silencing was undetectable in sugar-fed, blood-fed, and P. falciparum-infected female A. gambiae, while activation of the Toll pathway's Rel1 had a major impact. This study describes for the first time a single gene that influences an immune mechanism that is able to abort development of P. falciparum in Anopheline species. Further, this study addresses aspects of the molecular, evolutionary, and physiological consequences of the observed phenotype. These findings have implications for malaria control since broad-spectrum immune activation in diverse anopheline species offers a viable and strategic approach to develop novel malaria control methods worldwide. PMID- 19282972 TI - Detection of alpha-rod protein repeats using a neural network and application to huntingtin. AB - A growing number of solved protein structures display an elongated structural domain, denoted here as alpha-rod, composed of stacked pairs of anti-parallel alpha-helices. Alpha-rods are flexible and expose a large surface, which makes them suitable for protein interaction. Although most likely originating by tandem duplication of a two-helix unit, their detection using sequence similarity between repeats is poor. Here, we show that alpha-rod repeats can be detected using a neural network. The network detects more repeats than are identified by domain databases using multiple profiles, with a low level of false positives (<10%). We identify alpha-rod repeats in approximately 0.4% of proteins in eukaryotic genomes. We then investigate the results for all human proteins, identifying alpha-rod repeats for the first time in six protein families, including proteins STAG1-3, SERAC1, and PSMD1-2 & 5. We also characterize a short version of these repeats in eight protein families of Archaeal, Bacterial, and Fungal species. Finally, we demonstrate the utility of these predictions in directing experimental work to demarcate three alpha-rods in huntingtin, a protein mutated in Huntington's disease. Using yeast two hybrid analysis and an immunoprecipitation technique, we show that the huntingtin fragments containing alpha-rods associate with each other. This is the first definition of domains in huntingtin and the first validation of predicted interactions between fragments of huntingtin, which sets up directions toward functional characterization of this protein. An implementation of the repeat detection algorithm is available as a Web server with a simple graphical output: http://www.ogic.ca/projects/ard. This can be further visualized using BiasViz, a graphic tool for representation of multiple sequence alignments. PMID- 19282973 TI - Life, death, differentiation, and the multicellularity of bacteria. PMID- 19282974 TI - Ecological characterization of the colonic microbiota of normal and diarrheic dogs. AB - We used terminal restriction fragment polymorphism (T-RFLP) analysis to assess (1) stability of the fecal microbiota in dogs living in environments characterized by varying degrees of exposure to factors that might alter the microbiota and (2) changes in the microbiota associated with acute episodes of diarrhea. Results showed that the healthy canine GI tract harbors potential enteric pathogens. Dogs living in an environment providing minimal exposure to factors that might alter the microbiota had similar microbiotas; the microbiotas of dogs kept in more variable environments were more variable. Substantial changes in the microbiota occurred during diarrheic episodes, including increased levels of Clostridium perfringens, Enterococcus faecalis, and Enterococcus faecium. When diet and medications of a dog having a previously stable microbiota were changed repeatedly, the microbiota also changed repeatedly. Temporal trend analysis showed directional changes in the microbiota after perturbation, a return to the starting condition, and then fluctuating changes over time. PMID- 19282975 TI - The human vaginal bacterial biota and bacterial vaginosis. AB - The bacterial biota of the human vagina can have a profound impact on the health of women and their neonates. Changes in the vaginal microbiota have been associated with several adverse health outcomes including premature birth, pelvic inflammatory disease, and acquisition of HIV infection. Cultivation-independent molecular methods have provided new insights regarding bacterial diversity in this important niche, particularly in women with the common condition bacterial vaginosis (BV). PCR methods have shown that women with BV have complex communities of vaginal bacteria that include many fastidious species, particularly from the phyla Bacteroidetes and Actinobacteria. Healthy women are mostly colonized with lactobacilli such as Lactobacillus crispatus, Lactobacillus jensenii, and Lactobacillus iners, though a variety of other bacteria may be present. The microbiology of BV is heterogeneous. The presence of Gardnerella vaginalis and Atopobium vaginae coating the vaginal epithelium in some subjects with BV suggests that biofilms may contribute to this condition. PMID- 19282976 TI - Mitotic recombination: why? when? how? where? PMID- 19282977 TI - Genome-scale reconstruction of Escherichia coli's transcriptional and translational machinery: a knowledge base, its mathematical formulation, and its functional characterization. AB - Metabolic network reconstructions represent valuable scaffolds for '-omics' data integration and are used to computationally interrogate network properties. However, they do not explicitly account for the synthesis of macromolecules (i.e., proteins and RNA). Here, we present the first genome-scale, fine-grained reconstruction of Escherichia coli's transcriptional and translational machinery, which produces 423 functional gene products in a sequence-specific manner and accounts for all necessary chemical transformations. Legacy data from over 500 publications and three databases were reviewed, and many pathways were considered, including stable RNA maturation and modification, protein complex formation, and iron-sulfur cluster biogenesis. This reconstruction represents the most comprehensive knowledge base for these important cellular functions in E. coli and is unique in its scope. Furthermore, it was converted into a mathematical model and used to: (1) quantitatively integrate gene expression data as reaction constraints and (2) compute functional network states, which were compared to reported experimental data. For example, the model predicted accurately the ribosome production, without any parameterization. Also, in silico rRNA operon deletion suggested that a high RNA polymerase density on the remaining rRNA operons is needed to reproduce the reported experimental ribosome numbers. Moreover, functional protein modules were determined, and many were found to contain gene products from multiple subsystems, highlighting the functional interaction of these proteins. This genome-scale reconstruction of E. coli's transcriptional and translational machinery presents a milestone in systems biology because it will enable quantitative integration of '-omics' datasets and thus the study of the mechanistic principles underlying the genotype phenotype relationship. PMID- 19282978 TI - Robust detection and genotyping of single feature polymorphisms from gene expression data. AB - It is well known that Affymetrix microarrays are widely used to predict genome wide gene expression and genome-wide genetic polymorphisms from RNA and genomic DNA hybridization experiments, respectively. It has recently been proposed to integrate the two predictions by use of RNA microarray data only. Although the ability to detect single feature polymorphisms (SFPs) from RNA microarray data has many practical implications for genome study in both sequenced and unsequenced species, it raises enormous challenges for statistical modelling and analysis of microarray gene expression data for this objective. Several methods are proposed to predict SFPs from the gene expression profile. However, their performance is highly vulnerable to differential expression of genes. The SFPs thus predicted are eventually a reflection of differentially expressed genes rather than genuine sequence polymorphisms. To address the problem, we developed a novel statistical method to separate the binding affinity between a transcript and its targeting probe and the parameter measuring transcript abundance from perfect-match hybridization values of Affymetrix gene expression data. We implemented a Bayesian approach to detect SFPs and to genotype a segregating population at the detected SFPs. Based on analysis of three Affymetrix microarray datasets, we demonstrated that the present method confers a significantly improved robustness and accuracy in detecting the SFPs that carry genuine sequence polymorphisms when compared to its rivals in the literature. The method developed in this paper will provide experimental genomicists with advanced analytical tools for appropriate and efficient analysis of their microarray experiments and biostatisticians with insightful interpretation of Affymetrix microarray data. PMID- 19282979 TI - Natural transformation of helicobacter pylori involves the integration of short DNA fragments interrupted by gaps of variable size. AB - Helicobacter pylori are gram-negative bacteria notable for their high level of genetic diversity and plasticity, features that may play a key role in the organism's ability to colonize the human stomach. Homeologous natural transformation, a key contributor to genomic diversification, has been well described for H. pylori. To examine the mechanisms involved, we performed restriction analysis and sequencing of recombination products to characterize the length, fragmentation, and position of DNA imported via natural transformation. Our analysis revealed DNA imports of small size (1,300 bp, 95% confidence limits 950-1850 bp) with instances of substantial asymmetry in relation to selectable antibiotic-resistance markers. We also observed clustering of imported DNA endpoints, suggesting a possible role for restriction endonucleases in limiting recombination length. Additionally, we observed gaps in integrated DNA and found evidence suggesting that these gaps are the result of two or more separate strand invasions. Taken together, these observations support a system of highly efficient short-fragment recombination involving multiple recombination events within a single locus. PMID- 19282980 TI - Definition of herpes simplex virus type 1 helper activities for adeno-associated virus early replication events. AB - The human parvovirus Adeno-Associated Virus (AAV) type 2 can only replicate in cells co-infected with a helper virus, such as Adenovirus or Herpes Simplex Virus type 1 (HSV-1); whereas, in the absence of a helper virus, it establishes a latent infection. Previous studies demonstrated that the ternary HSV-1 helicase/primase (HP) complex (UL5/8/52) and the single-stranded DNA-Binding Protein (ICP8) were sufficient to induce AAV-2 replication in transfected cells. We independently showed that, in the context of a latent AAV-2 infection, the HSV 1 ICP0 protein was able to activate rep gene expression. The present study was conducted to integrate these observations and to further explore the requirement of other HSV-1 proteins during early AAV replication steps, i.e. rep gene expression and AAV DNA replication. Using a cellular model that mimics AAV latency and composite constructs coding for various sets of HSV-1 genes, we first confirmed the role of ICP0 for rep gene expression and demonstrated a synergistic effect of ICP4 and, to a lesser extent, ICP22. Conversely, ICP27 displayed an inhibitory effect. Second, our analyses showed that the effect of ICP0, ICP4, and ICP22 on rep gene expression was essential for the onset of AAV DNA replication in conjunction with the HP complex and ICP8. Third, and most importantly, we demonstrated that the HSV-1 DNA polymerase complex (UL30/UL42) was critical to enhance AAV DNA replication to a significant level in transfected cells and that its catalytic activity was involved in this process. Altogether, this work represents the first comprehensive study recapitulating the series of early events taking place during HSV-1-induced AAV replication. PMID- 19282981 TI - The tetraspanin protein CD37 regulates IgA responses and anti-fungal immunity. AB - Immunoglobulin A (IgA) secretion by plasma cells in the immune system is critical for protecting the host from environmental and microbial infections. However, the molecular mechanisms underlying the generation of IgA(+) plasma cells remain poorly understood. Here, we report that the B cell-expressed tetraspanin CD37 inhibits IgA immune responses in vivo. CD37-deficient (CD37-/-) mice exhibit a 15 fold increased level of IgA in serum and significantly elevated numbers of IgA(+) plasma cells in spleen, mucosal-associated lymphoid tissue, as well as bone marrow. Analyses of bone marrow chimeric mice revealed that CD37-deficiency on B cells was directly responsible for the increased IgA production. We identified high local interleukin-6 (IL-6) production in germinal centers of CD37-/- mice after immunization. Notably, neutralizing IL-6 in vivo reversed the increased IgA response in CD37-/- mice. To demonstrate the importance of CD37-which can associate with the pattern-recognition receptor dectin-1-in immunity to infection, CD37-/- mice were exposed to Candida albicans. We report that CD37-/- mice are evidently better protected from infection than wild-type (WT) mice, which was accompanied by increased IL-6 levels and C. albicans-specific IgA antibodies. Importantly, adoptive transfer of CD37-/- serum mediated protection in WT mice and the underlying mechanism involved direct neutralization of fungal cells by IgA. Taken together, tetraspanin protein CD37 inhibits IgA responses and regulates the anti-fungal immune response. PMID- 19282982 TI - Sm/Lsm genes provide a glimpse into the early evolution of the spliceosome. AB - The spliceosome, a sophisticated molecular machine involved in the removal of intervening sequences from the coding sections of eukaryotic genes, appeared and subsequently evolved rapidly during the early stages of eukaryotic evolution. The last eukaryotic common ancestor (LECA) had both complex spliceosomal machinery and some spliceosomal introns, yet little is known about the early stages of evolution of the spliceosomal apparatus. The Sm/Lsm family of proteins has been suggested as one of the earliest components of the emerging spliceosome and hence provides a first in-depth glimpse into the evolving spliceosomal apparatus. An analysis of 335 Sm and Sm-like genes from 80 species across all three kingdoms of life reveals two significant observations. First, the eukaryotic Sm/Lsm family underwent two rapid waves of duplication with subsequent divergence resulting in 14 distinct genes. Each wave resulted in a more sophisticated spliceosome, reflecting a possible jump in the complexity of the evolving eukaryotic cell. Second, an unusually high degree of conservation in intron positions is observed within individual orthologous Sm/Lsm genes and between some of the Sm/Lsm paralogs. This suggests that functional spliceosomal introns existed before the emergence of the complete Sm/Lsm family of proteins; hence, spliceosomal machinery with considerably fewer components than today's spliceosome was already functional. PMID- 19282983 TI - The nucleocapsid region of HIV-1 Gag cooperates with the PTAP and LYPXnL late domains to recruit the cellular machinery necessary for viral budding. AB - HIV-1 release is mediated through two motifs in the p6 region of Gag, PTAP and LYPX(n)L, which recruit cellular proteins Tsg101 and Alix, respectively. The Nucleocapsid region of Gag (NC), which binds the Bro1 domain of Alix, also plays an important role in HIV-1 release, but the underlying mechanism remains unclear. Here we show that the first 202 residues of the Bro1 domain (Bro(i)) are sufficient to bind Gag. Bro(i) interferes with HIV-1 release in an NC-dependent manner and arrests viral budding at the plasma membrane. Similar interrupted budding structures are seen following over-expression of a fragment containing Bro1 with the adjacent V domain (Bro1-V). Although only Bro1-V contains binding determinants for CHMP4, both Bro(i) and Bro1-V inhibited release via both the PTAP/Tsg101 and the LYPX(n)L/Alix pathways, suggesting that they interfere with a key step in HIV-1 release. Remarkably, we found that over-expression of Bro1 rescued the release of HIV-1 lacking both L domains. This rescue required the N terminal region of the NC domain in Gag and the CHMP4 binding site in Bro1. Interestingly, release defects due to mutations in NC that prevented Bro1 mediated rescue of virus egress were rescued by providing a link to the ESCRT machinery via Nedd4.2s over-expression. Our data support a model in which NC cooperates with PTAP in the recruitment of cellular proteins necessary for its L domain activity and binds the Bro1-CHMP4 complex required for LYPX(n)L-mediated budding. PMID- 19282984 TI - Measures of autozygosity in decline: globalization, urbanization, and its implications for medical genetics. AB - This research investigates the influence of demographic factors on human genetic sub-structure. In our discovery cohort, we show significant demographic trends for decreasing autozygosity associated with population variation in chronological age. Autozygosity, the genomic signature of consanguinity, is identifiable on a genome-wide level as extended tracts of homozygosity. We identified an average of 28.6 tracts of extended homozygosity greater than 1 Mb in length in a representative population of 809 unrelated North Americans of European descent ranging in chronological age from 19-99 years old. These homozygous tracts made up a population average of 42 Mb of the genome corresponding to 1.6% of the entire genome, with each homozygous tract an average of 1.5 Mb in length. Runs of homozygosity are steadily decreasing in size and frequency as time progresses (linear regression, p<0.05). We also calculated inbreeding coefficients and showed a significant trend for population-wide increasing heterozygosity outside of linkage disequilibrium. We successfully replicated these associations in a demographically similar cohort comprised of a subgroup of 477 Baltimore Longitudinal Study of Aging participants. We also constructed statistical models showing predicted declining rates of autozygosity spanning the 20th century. These predictive models suggest a 14.0% decrease in the frequency of these runs of homozygosity and a 24.3% decrease in the percent of the genome in runs of homozygosity, as well as a 30.5% decrease in excess homozygosity based on the linkage pruned inbreeding coefficients. The trend for decreasing autozygosity due to panmixia and larger effective population sizes will likely affect the frequency of rare recessive genetic diseases in the future. Autozygosity has declined, and it seems it will continue doing so. PMID- 19282985 TI - Genome-wide association analyses identify SPOCK as a key novel gene underlying age at menarche. AB - For females, menarche is a most significant physiological event. Age at menarche (AAM) is a trait with high genetic determination and is associated with major complex diseases in women. However, specific genes for AAM variation are largely unknown. To identify genetic factors underlying AAM variation, a genome-wide association study (GWAS) examining about 380,000 SNPs was conducted in 477 Caucasian women. A follow-up replication study was performed to validate our major GWAS findings using two independent Caucasian cohorts with 854 siblings and 762 unrelated subjects, respectively, and one Chinese cohort of 1,387 unrelated subjects--all females. Our GWAS identified a novel gene, SPOCK (Sparc/Osteonectin, CWCV, and Kazal-like domains proteoglycan), which had seven SNPs associated with AAM with genome-wide false discovery rate (FDR) q<0.05. Six most significant SNPs of the gene were selected for validation in three independent replication cohorts. All of the six SNPs were replicated in at least one cohort. In particular, SNPs rs13357391 and rs1859345 were replicated both within and across different ethnic groups in all three cohorts, with p values of 5.09 x 10(-3) and 4.37 x 10(-3), respectively, in the Chinese cohort and combined p values (obtained by Fisher's method) of 5.19 x 10(-5) and 1.02 x 10(-4), respectively, in all three replication cohorts. Interestingly, SPOCK can inhibit activation of MMP-2 (matrix metalloproteinase-2), a key factor promoting endometrial menstrual breakdown and onset of menstrual bleeding. Our findings, together with the functional relevance, strongly supported that the SPOCK gene underlies variation of AAM. PMID- 19282987 TI - Current status of the availability, development, and use of host plant resistance to nematodes. AB - Host plant resistance (HPR) to nematodes has been identified in many major crops and related wild germplasm. Most HPR is to the more specialized, sedentary endoparasitic genera and species, e.g., Globodera, Heterodera, Meloidogyne, Nacobbus, Rotylenchulus, and Tylenchulus. Some HPR has been developed or identified also to certain migratory endoparasites (Aphelenchoides, Ditylenchus, Pratylenchus, Radopholus) in a few hosts. Commercial use of HPR remains limited, despite its benefits to crop production when deployed appropriately. Restricted use and availability of HPR result from problems associated with transfer of resistance into acceptable cultivars. Difficulties occur in gene transfer to acceptable cultivars because of incompatibility barriers to hybridization or linkage to undesirable traits, for example in cucurbitaceous and solanaceous crops and sugarbeet. Specificity of HPR to only one species, or one or few pathotypes, as it relates to resistance durability and nematode virulence, and HPR response to abiotic factors such as high soil temperature, also limit availability and utility. A scheme for HPR development is presented to emphasize nematology research and information requirements for expanding HPR use in nematode control programs, for example in common bean, sugarbeet, and tomato. Nonbiological factors that influence HPR usage are discussed, including heavy reliance on nematicide programs, low priority of nematode HPR in many breeding programs, and insufficient breeder-nematologist collaboration. PMID- 19282988 TI - Problems and Strategies Associated with Long-term Use of Nematode Resistant Cultivars. AB - Plant-parasitic nematodes are obligate parasites, and planting cultivars that are highly resistant to these organisms places extensive selection pressure on the target species and affects nontarget nematodes as well. Problems encountered with long-term planting of cultivars resistant to nematodes include shifts in nematode races or species and the occurrence of multiple species of nematodes within the same field. These problems can be alleviated to some extent when crop management is used to lessen the selection pressure for change on the nematode populations. Race shifts within populations and possibly shifts between nematode species can be delayed by rotating susceptible cultivars and nonhost crops with resistant cultivars. Nematicides in conjunction with resistant cultivars may be used to limit damage by multiple species of nematodes. Some cultivars have resistance to multiple species of nematodes, but greatly increased research effort is needed in this area. More intensive plant breeding effort will be required to make nematode resistant cultivars competitive in quality and yield with more productive, susceptible cultivars. PMID- 19282986 TI - A novel role for MAPKAPK2 in morphogenesis during zebrafish development. AB - One of the earliest morphogenetic processes in the development of many animals is epiboly. In the zebrafish, epiboly ensues when the animally localized blastoderm cells spread, thin over, and enclose the vegetally localized yolk. Only a few factors are known to function in this fundamental process. We identified a maternal-effect mutant, betty boop (bbp), which displays a novel defect in epiboly, wherein the blastoderm margin constricts dramatically, precisely when half of the yolk cell is covered by the blastoderm, causing the yolk cell to burst. Whole-blastoderm transplants and mRNA microinjection rescue demonstrate that Bbp functions in the yolk cell to regulate epiboly. We positionally cloned the maternal-effect bbp mutant gene and identified it as the zebrafish homolog of the serine-threonine kinase Mitogen Activated Protein Kinase Activated Protein Kinase 2, or MAPKAPK2, which was not previously known to function in embryonic development. We show that the regulation of MAPKAPK2 is conserved and p38 MAP kinase functions upstream of MAPKAPK2 in regulating epiboly in the zebrafish embryo. Dramatic alterations in calcium dynamics, together with the massive marginal constrictive force observed in bbp mutants, indicate precocious constriction of an F-actin network within the yolk cell, which first forms at 50% epiboly and regulates epiboly progression. We show that MAPKAPK2 activity and its regulator p38 MAPK function in the yolk cell to regulate the process of epiboly, identifying a new pathway regulating this cell movement process. We postulate that a p38 MAPKAPK2 kinase cascade modulates the activity of F-actin at the yolk cell margin circumference allowing the gradual closure of the blastopore as epiboly progresses. PMID- 19282989 TI - Molecular transfer of nematode resistance genes. AB - Recombinant DNA techniques have been used to introduce agronomically valuable traits, including resistance to viruses, herbicides, and insects, into crop plants. Introduction of these genes into plants frequently involves Agrobacterium mediated gene transfer. The potential exists for applying this technology to nematode control by introducing genes conferring resistance to nematodes. Transferred genes could include those encoding products detrimental to nematode development or reproduction as well as cloned host resistance genes. Host genes that confer resistance to cyst or root-knot nematode species have been identified in many plants. The best characterized is Mi, a gene that confers resistance to root-knot nematodes in tomato. A map-based cloning approach is being used to isolate the gene. For development of a detailed map of the region of the genome surrounding Mi, DNA markers genetically linked to Mi have been identified and analyzed in tomato lines that have undergone a recombination event near Mi. The molecular map will be used to identify DNA corresponding to Mi. We estimate that a clone of Mi will be obtained in 2-5 years. An exciting prospect is that introduction of this gene will confer resistance in plant species without currently available sources of resistance. PMID- 19282990 TI - Breeding plants for resistance to nematodes. AB - Plant breeders and nematologists have developed improved cultivars of important crop species with resistance to plant-parasitic nematodes. The effectiveness of these breeding efforts has depended on the availability of efficient screening procedures, identification of adequate sources of durable resistance, nature of the nematode feeding habit, and knowledge of the inheritance of resistance. These factors determine to a large degree the breeding method and potential success of the research. Systematic searches for nematode resistance have identified resistant germplasm lines within crop species or from related species. When the resistance gene(s) is from related species, incongruity barriers or sterility of the resulting hybrids often must be overcome. In these situations, backcrossing is usually necessary to incorporate the resistance gene(s) and recover the desirable commercial traits of the crop species. If the resistance gene(s) is present within the crop species, the choice of breeding method depends on the inheritance of the resistance, type of screening procedure, and other important breeding objectives for the species. In the future, plant molecular biologists and geneticists will make available novel sources of nematode resistance through incorporation of transgenes from other genera. These efforts will likely require conventional breeding strategies before commercial utilization of an improved resistant cultivar. PMID- 19282991 TI - Evaluation of Nicotiana otophora as a Source of Resistance to Meloidogyne incognita Race 4 for Tobacco. AB - No currently available tobacco cultivar possesses resistance to Meloidogyne incognita race 4, nor has any source of resistance been reported within Nicotiana tabacum. The purpose of this study was to evaluate N. otophora acc. La Quinta as a source of resistance to this pathogen. Plants of tobacco cvs. NC 95 and NC 2326, N. otophora La Quinta and N. repanda were inoculated with second-stage juveniles of M. incognita race 4. Gall indices and egg-mass ratings were assessed at 4 and 8 weeks after inoculation. The two N. tabacum cultivars were heavily galled and had numerous egg masses at both rating periods. Nicotiana repanda was only weakly resistant. The galls on this species were very small and present at a low to moderate level; however, egg-mass ratings approaching those of the tobacco cultivars were observed 8 weeks after inoculation. In contrast, low gall indices and egg-mass ratings were found for N. otophora La Quinta at both the 4- and 8 week rating periods. In addition, little variability was observed within this species for either disease rating. Therefore, it appears that the La Quinta accession of N. otophora is a very promising source of M. incognita race 4 resistance for transfer to N. tabacum. PMID- 19282992 TI - Comparison of Reproduction by Meloidogyne graminicola and M. incognita on Trifolium Species. AB - The reproductive potential of Meloidogyne graminicola was compared with that of M. incognita on Trifolium species in greenhouse studies. Twenty-five Trifolium plant introductions, cultivars, or populations representing 23 species were evaluated for nematode reproduction and root galling 45 days after inoculation with 3,000 eggs of M. graminicola or M. incognita. Root galling and egg production by the two root-knot nematode species was similar on most of the Trifolium species. In a separate study, the effect of initial population densities (Pi) of M. graminicola and M. incognita on the growth of white clover (T. repens) was determined. Reproductive and pathogenic capabilities of M. graminicola and M. incognita on Trifolium spp. were similar. Pi levels of both root-knot nematode species as low as 125 eggs per 10-cm-d pots severely galled white clover plants after 90 days. Meloidogyne graminicola has the potential to be a major pest of Trifolium species in the southeastern United States. PMID- 19282993 TI - Impact of Thermal History on Tolerance of Meloidogyne hapla Second-stage Juveniles to External Freezing. AB - Low temperature induced physiological changes that increased the ability of second-stage juveniles of Meloidogyne hapla to survive external freezing. Second stage juveniles in polyethylene glycol solution were exposed to -4 , 0, 4, or 24 C, and then their survival was determined after ice-induced freezing of the suspensions at - 4 C for 24 hours. Survival was greatest for juveniles exposed to 4 C before freezing. Some juveniles were killed by exposure to - 4 C before freezing of the suspensions. The percentage of juveniles surviving freezing increased from about 30% to 80% within 12 hours of exposure to 4 C. This tolerance of external freezing was lost during subsequent exposure to 24 C. Longer exposures, of 1 to 15 days, to low temperature did not increase the percentage surviving external freezing, as compared to the 12-hour exposure, but reduced the tolerance of external freezing lost during subsequent exposure to 24 C for 48 hours. PMID- 19282994 TI - Routine Cryopreservation of Isolates of Steinernema and Heterorhabditis spp. AB - Infective-stage juveniles of Steinernema and Heterorhabditis spp. were cryopreserved using two-stage incubation in glycerol and 70% methanol before storage in cryotubes in liquid nitrogen. Optimal glycerol concentrations and incubation times for survival were determined for different species, but acceptable survival of all species and isolates of entomopathogenic nematodes can be obtained using 15% (w/w) glycerol and incubation for 48 hours. Mean survival was 69% for isolates of Steinernema and 68% for isolates of Heterorhabditis (n = 84). The maximum survival recorded was 97% for S. feltiae K254 stored in liquid nitrogen for 12 months. PMID- 19282995 TI - Comparison of assays for the determination of entomogenous nematode infectivity. AB - Injection, contact, and soil assays were used to compare infectivity of Heterorhabditis bacteriophora strain HP88 and Steinernema carpocapsae strain All to final instar Galleria mellonella larvae. Under comparable assay conditions, H. bacteriophora produced less Galleria mortality and showed greater within-assay variability in infectivity than S. carpocapsae. Injection of individual S. carpocapsae or H. bacteriophora infective juveniles into Galleria indicated that a comparatively greater percentage of S. carpocapsae was capable of initiating infection. In addition to nematode species, other major components of variability in assay estimations of nematode infectivity were number of nematodes used in the assay, assay type, date of the assay, and possibly, Galleria age. PMID- 19282996 TI - Modification of the Oligidic Medium for Axenic Culture of Aphelenchoides rutgersi. AB - Aphelenchoides rutgersi was axenically cultured in modified Soytone, yeast extract, lyophilized chick embryo extract medium (3% ST:2% YE:20% CEE-L, w/v:w/v:v/v). Earlier formulations used 10% CEE, v/v, before the manufacturer changed the preparation. After reestablishing A. rutgersi in medium that permitted continuous subcultivadon and reproduction, a second medium was tested that contained 0.5% sucrose and 0.5% Lipid Concentrate. The commercially available Lipid Concentrate made it possible to incorporate nonaqueous soluble chemicals into the medium. In addition, 0.1% Fast Green #3 was added to both media to visually demonstrate active ingestion of nutriment. PMID- 19282997 TI - Lectin Binding to Radopholus citrophilus and R. similis Proteins. AB - Lectin-binding glycoproteins in seven populations of two burrowing nematode sibling species were probed with five different biotinylated lectins on Western blots, and differences were correlated with nematode ability to parasitize citrus and to overcome citrus rootstock resistance. Banding patterns of molecular weight standards were fit best by an exponential decay function, and a predictive equation was used to estimate molecular weights (r(2) = 0.999). A band (131 kDa) that labeled with the lectin Concanavalin A (Con A) occurred in extracts from cuticles and egg shells of populations of Radopholus citrophilus that parasitize citrus. Wheat germ agglutin labeled a band (58 kDa) in aqueous homogenates of populations that reproduce in roots of citrus rootstock normally resistant to burrowing nematodes. The two sibling species R. citrophilus and R. similis were distinguished by a high molecular weight Con A-labeled band (608 kDa) from cuticle and egg shells. Probing blots with the lectin Limulus polyphemus agglutinin indicated that each population contained a band (12-16 kDa) specifically inhibited by the addition of 25 mM neuraminic acid, suggesting that glycoproteins with sialic acid moieties are present in burrowing nematodes. PMID- 19282998 TI - Detecting nematode features from digital images. AB - Procedures for estimating and calibrating nematode features from digitial images are described and evaluated by illustration and mathematical formulae. Technical problems, such as capturing and cleaning raw images, standardizing the grey level range of images, and the detection of characteristics of the body habitus, presence or absence of stylet knobs, and tail and lip region shape are discussed. This study is the first of a series aimed at developing a set of automated methods to permit more rapid, objective characterizations of nematode features than is achievable by cumbersome conventional methods. PMID- 19282999 TI - Description of Pratylenchus gutierrezi n. sp. (Nematoda: Pratylenchidae) from Coffee in Costa Rica. AB - A lesion nematode, Pratylenchus gutierrezi n. sp., collected from the roots of coffee in the Central Plateau of Costa Rica, is described and illustrated. Its relationships to Pratylenchus flakkensis, P. similis, and P. gibbicaudatus, the only other species of the genus having two head annules, males, or spermatheca with sperm, and an annulated tail terminus, is discussed. Other distinctive characters are its posterior vulva (mean of 80%); its prominently rounded stylet knobs, low head, and subcylindrical tail. SEM observations provide additional details of females and males, especially face views, which show for the first time sexual dimorphism. PMID- 19283000 TI - Morphometric Evidence for Three Juvenile Stages in Some Species of Xiphinema americanum sensu lato. AB - One to two hundred nematodes from each of seven Xiphinema americanum-group populations were measured to determine the range of stylet and body lengths for juveniles and adults. First-stage juveniles were identified by the position of the replacement odontostyle (i.e., the tip of the replacement odontostyle overlapped the base of the odontophore). Nematodes were identified as second stage if the functional odontostyle was the same length as the replacement odontostyle of the first stage. Subsequent stages were similarly identified by establishing the range of corresponding replacement and functional odontostyle lengths. In all populations examined, this procedure created natural divisions that clearly grouped nematodes by stylet and body length. Presumably these groups identified all juvenile and adult stages. Populations of X. americanum, X. rivesi, and X. californicum from the United States had three juvenile stages, but a population of X. pachtaicum from Bulgaria had four juvenile stages. PMID- 19283001 TI - Effect of Host Plant Age on Population Development and Pathogenicity of Ditylenchus destructor on Peanut. AB - The effect of inoculating peanut, Arachis hypogaea cv. Sellie, with Ditylenchus destructor at timed intervals after planting and with different initial nematode population densities (Pi) was tested in greenhouse experiments. Final nematode population densities (Pf) in hulls and seeds were greater (Pf < 0.001) in plants inoculated at or before 9 weeks after planting. Pod disease symptoms correlated positively with the Pf in the pods. The seedgrade of peanuts inoculated at or before 9 weeks after planting was reduced, whereas grade of peanuts from plants inoculated at 15 weeks or later was not reduced. Peanut plants inoculated 12 weeks after planting with a Pi of 10-100 had a lower Pf (P < 0.05) than plants with a Pi of 250 to 8,000. Seed of plants with a Pi of 250 or less could be marketed as choice edible seed, whereas those with a Pi of 500 or more were of reduced seedgrade. These results suggest that as few as 500 nematodes per plant at 12 weeks after planting can build up to injurious levels before harvest. A nematicide should therefore be active for longer than 12 weeks after planting to sufficiently suppress the population. PMID- 19283003 TI - Contemporary approaches to the study of host-parasite interactions: an introduction. PMID- 19283002 TI - Stem nematode-fusarium wilt complex in alfalfa as related to irrigation management at harvest time. AB - A high moisture level in the top 10 cm of soil at time of cutting of alfalfa increased the incidence of plant mortality and Fusarium wilt in soil infested with Ditylenchus dipsaci and Fusarium oxysporum f. sp. medicaginis in greenhouse and field microplot studies. Ranger alfalfa, susceptible to both D. dipsaci and F. oxysporum f. sp. medicaginis, was less persistent than Moapa 69 (nematode susceptible and Fusarium wilt resistant) and Lahontan alfalfa (nematode resistant with low Fusarium wilt resistance). In the greenhouse, the persistence of Ranger, Moapa 69, and Lahontan alfalfa plants was 46%, 64%, and 67% respectively, in nematode + fungus infested soil at high soil moisture at time of cutting. This compared to 74%, 84%, and 73% persistence of Ranger, Moapa 69, and Lahontan, respectively, at low soil moisture at time of cutting. Shoot weights as a percentage of uninoculated controls at the high soil moisture level were 38%, 40%, and 71% for Ranger, Moapa 69, and Lahontan, respectively. Low soil moisture at time of cutting negated the effect D. dipsaci on plant persistence and growth of subsequent cuttings, and reduced Fusarium wilt of plants in the nematode fungus treatment; shoot weights were 75%, 90%, and 74% of uninoculated controls for Ranger, Moapa 69, and Lahontan. Similar results were obtained in the field microplot study, and stand persistence and shoot weights were less in nematode + fungus-infested soil at the high soil-moisture level (early irrigation) than at the low soil-moisture level (late irrigation). PMID- 19283004 TI - Sex Determination in Caenorhabditis elegans. AB - In Caenorhabditis elegans, the decision to develop as a hermaphrodite or male is controlled by a cascade of regulatory genes. These genes and other tissue specific regulatory genes also control sexual fate in the hermaphrodite germline, which makes sperm first and then oocytes. In this review, we summarize the genetic and molecular characterization of these genes and speculate how they mutually interact to specify sexual fate. PMID- 19283005 TI - Molecular analysis of plant defense responses to plant pathogens. AB - A number of inducible plant responses are believed to contribute to disease resistance. These responses include the hypersensitive reaction, phytoalexin synthesis, and the production of chitinase, glucanase, and hydroxyproline-rich glycoproteins. Because of the coordinate induction of these responses, it has been difficult to determine whether they are functional defense responses, and if they are, how they specifically contribute to disease resistance. Recent developments in molecular biology have provided experimental techniques that will reveal the specific contribution of each response to disease resistance. In this paper, we describe a strategy to determine if the hypersensitive reaction is a functional plant defense mechanism. PMID- 19283006 TI - Molecular Analysis of the Interactions between Cyst Nematodes and Their Hosts. AB - In order to complete its life cycle, a cyst nematode must stimulate the production of a specialized syncytial feeding site within host root tissues. This process is characterized by major changes in local root morphology, including enlargement of affected nuclei and nucleoli, cell wall degradation, and proliferation of subcellular organelles. At the molecular level very little is known about the processes involved in this host response, but recent evidence suggests that cyst nematodes are able to regulate specific host genes. The host parasite model system provided by Arabidopsis thaliana and Heterodera schachtii will be fundamental to our future understanding of the formation of syncytia. Molecular biology now offers us the opportunity to study this complex host parasite interaction in great detail. A better understanding of the host genes regulated by cyst nematodes and the mechanisms by which this regulation is achieved will facilitate the engineering of crop cultivars that possess novel forms of resistance to these adept parasites. PMID- 19283007 TI - Random Amplified Polymorphic DNA Analysis of Heterodera cruciferae and H. schachtii populations. AB - Heterodera schachtii and H. cruciferae are sympatric in California and frequently occur in the same field upon the same host. We have investigated the use of polymerase chain reaction (PCR) amplification of nematode DNA sequences to differentiate H. schachtii and H. cruciferae and to assess genetic variability within each species. Single, random oligodeoxyribonucleotide primers were used to generate PCR-amplified fragments, termed RAPD (random amplified polymorphic DNA) markers, from genomic DNA of each species. Each of 19 different random primers yielded from 2 to 12 fragments whose size ranged from 200 to 1,500 bp. Reproducible differences in fragment patterns allowed differentiation of the two species with each primer. Similarities and differences among six different geographic populations of H. schachtii were detected. The potential application of RAPD analysis to relationships among nematode populations was assessed through cluster analysis of these six different populations, with 78 scorable markers from 10 different random primers. DNA from single cysts was successfully amplified, and genetic variability was revealed within geographic populations. The use of RAPD markers to assess genetic variability is a simple, reproducible technique that does not require radioisotopes. This powerful new technique can be used as a diagnostic tool and should have broad application in nematology. PMID- 19283008 TI - Life Cycle and Reproductive Potential of the Nematode Heterorhabditis bacteriophora Strain HP88. AB - Development of the entomopathogenic nematode Heterorhabditis bacteriophora strain HP88 was studied in vivo with larvae of the greater wax moth, Galleria mellonella, as host and in vitro. At 25 C in vivo, the duration of the life cycle from egg hatch to egg hatch was 96 hours. Juvenile development took 48 hours, with the duration of each juvenile stage ranging from 8 to 12 hours. Under crowded conditions, development proceeded to the infective juvenile (IJ) stage instead of the third juvenile stage (J3). Life-cycle duration and proportion of the various developmental stages in the population were similar in in vitro and in vivo cultures. When in vivo or in vitro development was initiated from the IJ stage, only hermaphrodites developed in the first generation and males appeared only in the second generation. The average (+/-SD) number of progeny per hermaphrodite was 243 +/- 98. The ratio of males to hermaphrodites in the second generation was 1:9.4 +/- 6.8. PMID- 19283009 TI - Phenotypic and Genetic Analysis of a Mutant of Heterorhabditis bacteriophora Strain HP88. AB - Induction and characterization of a morphological mutant are described for Heterorhabditis bacteriophora strain HP88. A homozygous inbred line was used as the base population for mutagenesis and genetic analysis of mutations. Mutagenesis was induced by exposing young hermaphrodites to 0.05 M ethyl methanesulfonate. A dumpy mutant (designated Hdpy-l) was isolated from the F generation of the mutagenized population. Morphological studies with light and scanning electron microscopy revealed that the head region of the adult stage was compressed. The head region of the infective juvenile was distorted and the mouth open. Backcross with the original population was successful only between mutant hermaphrodites and wild type males; 50-100 percent of the progeny of this cross maintained the dumpy phenotype, indicating that the ratio between self- and external fertilization of the eggs is > 1 and that the dumpy mutation is recessive. PMID- 19283010 TI - Effect of exsheathment on motility and pathogenicity of two entomopathogenic nematode species. AB - The effect of sheath loss on motility and pathogenicity of the entomopathogenic nematodes, Heterorhabditis bacteriophora and Steinernema carpocapsae, was examined using both naturally and chemically exsheathed (desheathed) infective juveniles. Exsheathed S. carpocapsae showed increased motility on agar compared to sheathed nematodes. The presence of a host increased motility threefold in all S. carpocapsae treatments. These results suggest that activation of S. carpocapsae host finding may result from sheath loss in addition to host stimuli. Desheathed H. bacteriophora were significantly less motile than the sheathed or exsheathed groups. The decreased motility may be due to adverse effects of the chemical treatment for desheathment. Sheath loss did not affect the pathogenicity of either species. PMID- 19283011 TI - Use of a Stilbene Brightener, Tinopal LPW, as a Radiation Protectant for Steinernema carpocapsae. AB - A stilbene fluorescent brightener, Tinopal LPW, was used as an ultraviolet (UV) protectant for the entomopathogenic nematode Steinernema carpocapsae (All strain). Irradiation of an aqueous suspension of nematodes produced a LC in 15.7 minutes under a sunlamp and in 31.7 minutes in direct sunlight. Irradiation by both sunlamp and sunlight of a suspension of nematodes in Tinopal LPW did not reduce their biological activity as measured by their ability to parasitize wax moth larvae after exposure of 8 hours and 4 hours, respectively. Tinopal LPW appeared promising as a radiation protectant. PMID- 19283012 TI - Field Application of Entomopathogenic Nematodes for Control of Delia radicum in Collards. AB - Control of Delia radicum (cabbage maggot) in field collards (Brassica oleracea) was compared after one or two applications of entomopathogenic nematodes, Steinernema carpocapsae (All strain) and Heterorhabditis bacterophora (HP88 strain), a single application of granular chlorpyrifos, and a water-only treatment. Nematodes were applied with a sprayer during the egg stage of first generation D. radicum, and chlorpyrifos was hand placed around collard stems during the same period. A second nematode application was made 10 days later. Chlorpyrifos treatment resulted in fewer puparia per plant, less root damage and higher yield than all other treatments, including the control. Collard yield from nematode-treated beds did not differ from controls. These data indicate that, under these field conditions, the species or strains of entomopathogenic nematodes tested did not reduce the number of active cabbage maggots, nor did they prevent collard root damage. PMID- 19283013 TI - New species of Rhabdochona Railliet, 1916 (Nemata: Rhabdochonidae) from Rainbow Trout in California Streams. AB - Three new species of Rhabdochona Railliet, 1916 are described and illustrated from Salmo gairdneri Richardson (rainbow trout) in freshwater streams in California: Rhabdochona californiemis n. sp., R. paxmani n. sp., and R. satmonis n. sp. Rhabdochona californiensis n. sp. is characterized by 14 anteriorly directed teeth in the prostome, egg devoid of filaments or floats, male and female tail terminus with a single mucro, left (long) spicule slender with a moderate distended podoid terminal end, spicular ratio 1:3.8. Rhabdochona paxmani n. sp. is characterized by 10 teeth in the prostome, eggs with polar floats, left (long) spicule slender with podoid terminus distended and having a minute subterminal spine; right spicule with prominent gorgeret (barb), spicular ratio 1:4.3, male and female tail terminus with a cuticular conical rounded short projection. Rhabdochona salmoni, n. sp. is characterized by 10 teeth anteriorly directed in the prostome, eggs with polar floats, left spicule slender with a distended podoid terminus; right spicule with a sharply indented gorgeret, spicular ratio 1:4.3, male and female tail terminus with a conical or rounded tip. PMID- 19283014 TI - Taxonomy of Discocriconemella (Nematoda: Criconematoidea) with a Redescription of D. mauritiensis. AB - The form of the cephalic disc and its taxonomic significance at the species level in the genus Discocriconemella De Grisse &Loof, 1965 is discussed. By light (LM) and scanning electron microscopy (SEM), four groups of disc configurations in females and juveniles are distinguishable. The disc is either round with an uninterrupted margin (group 1), has a deep dorsal and ventral indentation (group 2), is indented medially and laterally giving a four-lobed appearance (group 3), or is round with paired dorsal and ventral projections (group 4). There is no apparent correlation between groups of cephalic discs and other characters such as tail shape or number of body annules. Discocriconemella mauritiensis (Williams, 1960) De Grisse &Loof, 1965 is redescribed from a sugar cane population collected in Mauritius, and the diagnosis of the genus is emended. PMID- 19283015 TI - Morphometric Variation and Biogeography of Ogwa menzeli and Criconema sphagni. AB - Morphometrics of Ogma menzeli from woodlands in the Adirondack Mountains of New York State and in Iowa were compared. Specimens from the Adirondacks were significantly greater in mean total body length, stylet length, the b, R, and RV values, body width, and esophagus length than specimens from Iowa. The V value was significantly greater in the Iowa than in the Adirondack specimens. The two populations are considered ecotypes of O. menzeli. Criconema sphagni morphometric measurements differed significantly for the RV value (negative) and V value (positive) relative to elevation in the Adirondacks. There was a positive regression correlation for the RV value of O. menzeli and elevation in the Adirondack Mountains. PMID- 19283016 TI - Variability among Populations of Meloidogyne arenaria. AB - Variability in reproduction and pathogenicity of 12 populations of Meloidogyne arenaria race 1 was evaluated on Florunner peanut, Centennial soybean, Rutgers tomato, G70, K326, and Mc944 tobacco, and Carolina Cayenne, Mississippi Nemaheart, and Santanka pepper. Differences among M. arenaria populations in rates of egg production 45 days after inoculation were observed for all cultivars except Santanka pepper. Differences among populations in dry top weights or fresh root weights were recorded on all cultivars. Numbers of nematode eggs produced on Florunner peanut varied from 3,419 to 11,593/g fresh root weight. On resistant tobacco cultivars (G70 and K326), one nematode population produced high numbers of eggs (12,042 and 6,499/g fresh root weight on G70 and K326, respectively), whereas the other populations produced low numbers of eggs (less than 500 eggs/g fresh root weight on both cultivars). Two variant M. arenaria race 1 populations were identified by factor analysis of reproductive rates on all nine cultivars. Differences m reproduction and pathogenicity observed among populations would affect the design of sustainable management systems for M. arenaria. PMID- 19283017 TI - Fitness Components and Selection of Biotypes of Heterodera glycines. AB - Survival of biotypes of Heterodera glycines was studied in microplots and in the field. The field population was subjected to various cropping sequences. Viability of eggs overwintered in microplots was determined each spring by percentage hatch, percentage of hatched eggs penetrating roots, and numbers of females developing on Peking and PI 88788 soybeans. Eggs from the field were collected in the spring and fall and assayed for ability to develop on Peking and PI 88788. Hatch of isolates overwintered in the microplots averaged 13% in May 1989 and 19% in 1990. No differences in hatch were detected among the isolates in 1989. Numbers of juveniles penetrating susceptible roots averaged less than 20% of the hatched eggs each year. An isolate of a biotype parasitic on susceptible soybeans and the resistant soybean PI 88788 penetrated roots more successfully than other biotypes. A second isolate from North Carolina, parasitic on susceptible soybeans, PI 88788, and the resistant soybean Peking experienced selection against development on Peking during two winters. Only 17 % of the expected numbers of females developed on Peking from this isolate. In the microplot experiment, parasitism of PI 88788 and Peking had a selective disadvantage (selection coefficient) of s = 0.29 and 0.62 over all isolates, respectively. In the field experiment, the relative numbers of cysts on Peking and PI 88788 increased between the spring and fall on soybean, then decreased over the winter and under corn. Selection coefficients against parasitism of PI 88788 and Peking averaged 0,19 and 0.3 in the field population. In neither experiment did juveniles lose their ability to parasitize susceptible soybeans. PMID- 19283018 TI - Predisposition of Broadleaf Tobacco to Fusarium Wilt by Early Infection with Globodera tabacum tabacum or Meloidogyne hapla. AB - In greenhouse experiments, broadleaf tobacco plants were inoculated with tobacco cyst (Globodera tabacum tabacum) or root-knot (Meloidogyne hapla) nematodes 3, 2, or 1 week before or at the same time as Fusarium oxysporum. Plants infected with nematodes prior to fungal inoculation had greater Fusarium wilt incidence and severity than those simultaneously inoculated. G. t. tabacum increased wilt incidence and severity more than did M. hapla. Mechanical root wounding within 1 week of F. oxysporum inoculation increased wilt severity. In field experiments, early-season G. t. tabacum control by preplant soil application of oxamyl indirectly limited the incidence and severity of wilt. Wilt incidence was 48%, 23%, and 8% in 1989 and 64%, 60%, and 19% in 1990 for 0.0, 2.2, and 6.7 kg oxamyl/ha, respectively. Early infection of tobacco by G. t. tabacum predisposed broadleaf tobacco to wilt by F. oxysporum. PMID- 19283019 TI - Dynamics of Belonolaimus longicaudatus Parasitism on a Susceptible St. Augustinegrass Host. AB - St. Augustinegrass (Stenotaphrum secundatum) cv FX-313 was used as a model laboratory host for monitoring population growth of the sting nematode, Belonolaimus longicaudatus, and for quantifying the effects of sting nematode parasitism on host performance in two samples of autoclaved native Margate fine sand with contrasting amounts of organic matter (OM = 7.9% and 3.8%). Following inoculation with 50 Belonolaimus longicaudatus per pot, nematodes peaked at a mean of 2,139 nematodes per pot 84 days after inoculation, remained stable through 168 days at 2,064 nematodes per pot, and declined at 210 days. The relative numbers of juveniles and adults demonstrated senescence after 84 days. Root dry weight of nematode-inoculated plants increased briefly to an apparent equilibrium 84 days after inoculation, whereas root weights of uninoculated controls continued to increase, exceeding those of inoculated plants from 84 to 210 days (P < 0.01). At 210 days, uninoculated plants had 227% the root dry weight of inoculated plants. Transpiration of FX-313 was reduced by nematodes (P < 0.0001) at 84 and 126 days after inoculation; reduction was first observed at 42 days and last observed 168 days after inoculation (P < 0.05). OM content affected all plant performance variables at multiple dates, and generally there were no inoculation x OM content interactions. OM content had no effect on nematode numbers per pot, although there was a slight (P < 0.05) increase in the number of nematodes per gram root dry weight in the low-OM soil compared with the high-OM soil. PMID- 19283020 TI - Damage Potential and Reproduction of Belonolaimus longicaudatus and Hoplolaimus galeatus on Alyceclover. AB - Alyceclover (Alysicarpus spp.) is an annual, high-quality leguminous forage, suitable for production under tropical and subtropical climates where the husbandry of conventional leguminous forages, Trifolium spp., is uneconomical. The damage potential and reproduction of Belonolairaus longicaudatus and Hoplolaimus galeatus on alyceclover were studied under greenhouse conditions, using sand and sandy clay loam soil materials, respectively. Both nematode species reproduced on alyceclover, but only B. longicaudatus was pathogenic. Symptoms of B. longicaudatus damage were suppression of shoot yield, limited root system, stunting, incipient wilting, and occasional seedling mortality. In one experiment, the threshold-damage density was three nematodes/100 cm(3) sand, whereas in the other experiment it was zero nematodes. PMID- 19283021 TI - Pathological Effects of Pratylenchus neglectus on Wheatgrasses. AB - In controlled greenhouse and growth chamber studies, Pratylenchus neglectus reduced dry shoot and dry root weight of rangeland grasses. Greenar intermediate wheatgrass and Secar Snake River wheatgrass were more susceptible to P. neglectus than Hycrest crested wheatgrass, Fairway crested wheatgrass, and Nordan crested wheatgrass at a greenhouse bench temperature of 26 +/- 3 C. Hycrest was the most tolerant to parasitism by P. neglectus. An initial nematode inoculum density of four nematodes/cm(3) soil reduced dry shoot weights of Hycrest, Fairway, Nordan, Greenar, and Secar by 22%, 33%, 36%, 47%, and 49%, and reduced dry root weights by 26%, 31%, 32%, 38%, and 42%. There was a positive relationship between dry root weight, the nematode inoculum density, and the nematode reproduction index (final nematode population/initial nematode inoculum). However, there were more nematodes/g root tissue on Secar than on the crested wheatgrasses, and significantly more nematodes/g root tissue on Greenar, Fairway, and Nordan than on Hycrest. Pratylenchus neglectus was most pathogenic at four nematodes/cm(3) soil at 30 C and least pathogenic at one nematode/cm(3) soil at 15 C. Greenar and Secar were more susceptible to the nematode than Hycrest, Fairway, and Nordan at two and four nematodes/cm(3) soil at 20 to 30 C. The nematode reproductive indices were greatest at 30 C and were positively correlated with dry root weight. Secar supported the most and Hycrest had the fewest nematodes/g root. PMID- 19283022 TI - Effects of Cropping Sequences on Population Densities of Meloidogyne hapla and Carrot Yield in Organic Soil. AB - The influence of various cropping sequences on population densities of Meloidogyne hapla and carrot yield was studied in organic soil under microplot and field conditions. Spinach, radish, barley, oat, and wheat were poor or nonhosts for M. hapla. Population densities of M. hapla were maintained or increased on cabbage, celery, lettuce, leek, marigold, and potato. Marketable percent-age and root weight of carrots were greater following spinach, oat, radish, and fallow-onion than those following two crops of onion or carrot in microplots. Under field conditions, the carrot-onion-oat-carrot cropping sequence decreased M. hapla population densities and provided a 282% increase in marketable yield of carrot compared to a carrot monoculture. Two consecutive years of onion increased M. hapla population densities causing severe root galling and a 50% yield loss in the following crop of carrot. Based on root-gall indices, carrots could be grown economically for 2 years following radish, spinach, and oat, but not following onion and carrot without the use of nematicides. PMID- 19283023 TI - A plant health care program for brambles in the pacific northwest. AB - Pratylenchus and Xiphinema species have been associated with decline and mortality of brambles (Rubus species) in the Pacific Northwest of the United States. These nematodes cause direct feeding damage and (or) transmit viruses that result in poor fruit quality and plant decline. A nematode management program has been developed by the author to minimize chemical use and nematode induced damage while optimizing fruit production. Nematode management is an integral part of a total plant health care program in which foliar and soil pests, plant stresses, and fertility are managed. PMID- 19283024 TI - Steinernema neocurtillis n. sp. (Rhabditida: Steinernematiclae) and a Key to Species of the Genus Steinernema. AB - Steinernema neocurtillis n. sp. isolated from the mole cricket Neocurtilla hexadactyla Perty can be distinguished from other members of the genus by characteristics of the first-generation male and the third-stage infective juvenile (IJ). In the male, the distance from the anterior end to the excretory pore (DAE) is less than the body width at the excretory pore; D% (DAE divided by length of esophagus x 100) is low at 19. The gubernaculum legth is greater than three-fourths the spicule length. Range of the ratio gubernaculum length divided by spicule length is 0.82-0.93 in the first-generation male and 0.92-1.00 in the second-generation male. In the IJ, the distance from the anterior end to the excretory pore is extremely short (18 mum), causing the D% and E% (DAE divided by tail length x 100) to be low (D% = 23 and E% = 12). Average body length of the IJ is 885 mum. PMID- 19283025 TI - Effects of Enzymes, Chemicals, and Tempertaure on Steinernema carpocapsae Attraction to Host Plasma. AB - Migration of exsheathed infective juveniles of Steinernema carpocapsae to plasma of the host insect Spodoptera litura was not affected by treatments with the lectins concanavalin A, soybean agglutinin, or wheat germ agglutinin; with the enzymes neuraminidase, alpha-mannosidase, lipase, pronase, or phospholipase C; or with cetyl trimethylammonium bromide or spermidine. Treatment with sodium metaperiodate or sodium hypochlorite inhibited nematode attraction towards insect plasma; numbers of randomly wandering nematodes increased. Nematode migration towards the source of attraction was unaffected by temperatures below 33 C but was impaired at 35 and 37 C. The adverse effect of 5 mM and 10 mM NaIO on migratory behavior was reversed 24 hours after rinsing with buffered saline. The effect of NaOCl on nematode behavior was slightly reversible at concentrations of 0.2 and 0.4% (v/v) but apparently irreversible at 0.6 and 1.0%. The effect of heat treatment at 35 and 37 C was reversible. PMID- 19283026 TI - Within-wood Spatial Dispersion of the Pinewood Nematode, Bursaphelenchus xylophilus. AB - Pinewood nematode, Bursaphelenchus xylophilus (Steiner &Buhrer) Nickle, spatial dispersion was determined in Scots pine, Pinus sylvestris L., bolts infested with the pine sawyer beetle, Monochamus carolinensis (Olivier) and in bolts without M. carolinensis. According to Taylor's power law and Green's index of dispersion, nematode dispersion was aggregated in both sets of bolts. The degree of aggregation did not differ significantly between beetle-infested and noninfested bolts, suggesting that the presence of M. carolinensis does not affect nematode dispersion within a bolt. Nematode population densities differed radially in bolts not infested with pine sawyers, but in a nonregular pattern. Moisture content of the bolts was correlated with population density of B. xylophilus, suggesting that nematode aggregates occur in areas of high moisture content. PMID- 19283027 TI - Survival and Infectivity of Bursaphelenchus xylophilus in Wood Chip-Soil Mixtures. AB - To determine the effect of soil environment on the life stages and total numbers of Bursaphelenchus xylophilus, nematode-infested wood chips alone and mixed with soil were incubated at 12 and 20 C. Nematodes were extracted at 2-week intervals for 12 weeks. Numbers of nematodes and percentage of third-stage dispersal larvae were greater at 12 C and in chips without soil. Percentage of juveniles of the propagative cycle was greater at 20 C and in chips with soil. Although B. xylophilus survived in chips with soil for 12 weeks, nematode numbers and life stage percentages changed little over time. To determine if B. xylophilus was capable of infecting wounded roots, infested and uninfested chips were mixed with soil in pots with white and Scots pine seedlings. Trees were maintained at 20 and 30 C and harvested at mortality or after 12 weeks. Only seedlings treated with infested chips contained nematodes. In field experiments, planted seedlings were mulched with infested chips to determine if nematodes would invade basal stem wounds. Among these trees, Scots pine was more susceptible than white or red pines to infection and mortality. PMID- 19283028 TI - Effects of Temperature on Pratylenchus neglectus and on Its Pathogenicity to Barley. AB - In a petri-dish study, development of the nematode Pratylenchus neglectus was observed every 4 days, and stage-specific development times were estimated, using a parameter estimation algorithm for a distributed-delay population model. The lower threshold temperature for development of a population of P. neglectus was 7.75 C. Temperatures above 25 C were unfavorable for this population on barley. Total numbers of P. neglectus in barley roots and associated soil in pots were greatest at 25 C and lower at temperatures above and below that level. There was no change in nematode numbers per gram of root as temperature increased between 24 C and 32 C because root weights decreased at higher temperatures. Restricted root mass may contribute to the lower total nematode population levels at higher temperature. Maximum number of nematodes moved through a 2-cm layer of sand on a Baermann funnel at about 20 C; lowest number of nematodes moved at 10 C and 30 C. PMID- 19283029 TI - Effect of Temperature on Attachment, Development, and Interactions of Pasteuria penetrans on Meloidogyne incognita. AB - The effect of temperature (10, 20, 25, 30, and 35 C) on attachment and development of Pasteuria penetrans on Meloidogyne arenaria race 1 was elevated in growth chambers. The greatest attachment rate of endospores of P. penetrans occurred on second-stage juveniles at 30 C. The bacterium developed more quickly within its host at 30 and 35 C than at 25 C or below. The development of the bacterium within the nematode female was divided into nine recognizable life stages, which ranged from early vegetative thalli to mature sporangia. Mature sporangium was the predominant life stage observed after 35, 40, 81, and 116 days at 35, 30, 25, and 20 C, respectively. The body width and length of M. arenaria females infected with P. penetrans were smaller initially than the same dimensions in uninfected females, but became considerably larger over time at 25, 30, and 35 C. This isolate of P. penetrans also parasitized and completed its life cycle in males of M. arenaria. PMID- 19283030 TI - The Role of Microbes Associated with Chicken Litter in the Suppression of Meloidogyne arenaria. AB - The role of microbes associated with chicken litter in the suppression of Meloidogyne arenaria in amended soil was investigated. Amended soil treatments were prepared, including combinations of sterile and nonsterile chicken litter and soil. Microbial biomass in different treatments was compared by measuring carbon dioxide evolution. There was less CO evolved in sterile litter than in nonsterile litter treatments. Tomato seedlings cv. Rutgers were transplanted into soil mixtures and inoculated with 2,000 M. arenaria eggs. After 10 days, fewer second-stage juveniles (J2) had penetrated the roots in soils amended with nonsterile litter than sterile litter. The effects of sterile and nonsterile litter-amended soil solutions on M. arenaria eggs and J2 were observed over a period of 6 days. A lower percentage of eggs remained apparently healthy in nonsterile than in sterile-amended soil solutions over 6 days. Microbial degradation of the egg shells was apparent. Fewer J2 survived in sterile- and nonsterile-amended-soil solutions as compared to water controls. PMID- 19283031 TI - Minimizing damage by Ditylenchus destructor to Peanut Seed with Early Harvest. AB - Greenhouse and microplot experiments were conducted to evaluate the damage potential of Ditylenchus destructor on four South African commercial peanut cultivars as influenced by harvest date. The cultivars Sellie and Harts should be harvested by 150 and 120 days after planting, respectively. Losses were 12-13% with early harvest, but a 15-day delay resulted in losses of 45-49%. Harvest of Natal Common and Norden at 125 and 145 days after planting, respectively, resulted in the highest seed grade. By normal harvest time (140 and 160 days, respectively) these two cultivars were downgraded to crushing seed quality. Even though seed weight increases with time, a net loss occurs if harvest is delayed. PMID- 19283032 TI - Nematode numbers and crop yield in a fenamiphos-treated sweet corn-sweet potato vetch cropping system. AB - Nematode population densities and yield of sweet corn and sweet potato as affected by the nematicide fenamiphos, in a sweet corn-sweet potato-vetch cropping system, were determined in a 5-year test (1981-85). Sweet potato was the best host of Meloidogyne incognita of these three crops. Fenamiphos 15G (6.7 kg a.i./ha) incorporated broadcast in the top 15 cm of the soil layer before planting of each crop increased (P 1.5. These included almond (Desmayo Rojo, 1143), apple (EM-9, EM-106), avocado (Hass), cherry (Santa Lucia 64, Camil, M x M 14, Masto de Montafiana), grape (41-B, Fercal, Ritcher 110), hazelnut (Pauetet), loquat (Nadal), peach (Montclar, GF-305), pear (OHF-333), pistachio (P. atlantica, P. vera, P. terebinthus), plum (San Julian 655-2, Montizo, Pixy, Myrobalan 605), and walnut (Serf). The peach rootstock Nemaguard and the grape 161-49 had Pf/Pi between 1.0 and 1.5 (slightly higher than inoculation level). All the tested citrus (Alemow, rough lemon, Carrizo citrange, sour orange, Troyer citrange, Citrumelo), plus three grape (SO4, Vitis rupestris, 1103-P), and the olive rootstock Arbequina had a Pf/Pi < 1.0. PMID- 19283048 TI - Comparison of nematode population densities on six summer crops at seven sites in north Florida. AB - Densities of plant-parasitic nematodes were compared on six crops grown for forage during the summer of 1991 at seven sites in north central Florida. The cropping treatments were 'Howard' soybean (Glycine max), 'Deltapine 105' soybean, velvetbean (Mucuna deeringiana), 'California Blackeye #5' cowpea (Vigna unguiculata), 'Pioneer 3098' tropical corn (Zea mays), and 'Asgrow Chaparral' sorghum (Sorghum bicolor). Highest final densities (Pf) of Meloidogyne incognita and Criconemella spp. were obtained following corn or sorghum at most sites. The lowest Pf of M. incognita occurred after velvetbean at all seven sites, but Pf after cowpea were equivalent to Pf after velvetbean at four of seven sites. Cultivar choice is critical in planning rotations to suppress M. incognita because results obtained here and elsewhere have shown great differences among sorghum and cowpea cultivars. The Pf of Pratylenchus spp. were lowest following velvetbean at four of seven sites. There were no differences in densities of Paratrichodorus minor among crops, but populations increased at a greater rate if initial density (Pi) was low. Multiplication rates (Pf/Pi) of most nematode species on most crops varied inversely with Pi. An accurate impression of nematode multiplication and host status could not be obtained unless a range of Pi was examined. PMID- 19283049 TI - Effects of Root Decay on the Relationship between Meloidogyne spp. Gall Index and Egg Mass Number in Cucumber and Horned Cucumber. AB - A greenhouse study was conducted to determine if root necrosis had an effect on the relationship between root-knot nematode gall index and egg mass number. Thirty-four cultigens of Cucumis (14 accessions, 12 cultivars, and six breeding lines of C. sativus, and two accessions of C. metuliferus) were evaluated against four root-knot species (Meloidogyne arenaria race 2, M. incognita race 1, M. incognita race 3, and M. javanica) measuring gall index, root necrosis, and egg mass number. Root necrosis affected the gall index-egg mass relationship. At lower root necrosis values, a stronger relationship existed between gall index and egg mass number than at higher root necrosis values. Root tissue was destroyed by root necrosis, and normal root-knot nematode reproduction would not occur, even though root galling was still observed. The races of M. incognita tested had a greater effect in predisposing C. sativus and C. metuliferus to root necrosis than did M. arenaria race 2 or M. javanica. This study showed that root necrosis had an adverse affect on the relationship between gall index and egg mass number in cucumber. PMID- 19283050 TI - Field Evaluation of Susceptibility to Meloidogyne arenaria in Arachis hypogaea Plant Introductions. AB - Resistance to Meloidogyne arenaria race 1 is not currently available in commercial peanut cultivars. Moderate levels of resistance have been identified in Arachis hypogaea plant introductions (PI) in previous greenhouse studies. The purpose of this work was to evaluate the effects of resistance in peanut PI on populations dynamics of M. arenaria in field plots. The PI designated as resistant in greenhouse studies had fewer M. arenaria in roots than the most susceptible PI. At midseason and at the end of the season, resistant PI had fewer M. arenaria in rhizosphere soil than the most susceptible PI. Seven resistant PI had lower numbers of M. arenaria than 'Florunner' at the end of the growing season. Gall index, egg mass index, number of eggs/plant, and number of eggs/g root from greenhouse screening were highly correlated with population levels of M. arenaria in the field, especially at midseason. These greenhouse indices should provide reliable estimates of host suitability in future studies. PMID- 19283051 TI - Reproduction of Four Races of Meloidogyne incognita on Hibiscus cannabinus. AB - The feasibility of cultivation of kenaf (Hibiscus cannabinus) in the United States is receiving a multifaceted evaluation. Among the factors being evaluated is kenafs susceptibility to nematodes. In this investigation, four races of Meloidogyne incognita reproduced extensively on each of the several kenaf genotypes examined in greenhouse tests. Some genotypes of kenaf, however, demonstrated limited resistance to certain races of M. incognita. PMID- 19283052 TI - Host suitability of Ixora spp. for the Root-knot Nematodes Meloidogyne incognita Race 1 and M. javanica. AB - Eight commonly cultivated Ixora species or cultivars were tested for their suitability as hosts and their level of tolerance to Meloidogyne incognita race 1 and M. javanica in a greenhouse study. Twenty weeks postinoculation with 5,000 eggs per pot, M. incognita race 1 and M. javanica produced galls and formed egg masses on roots of all eight Ixora species or cultivars tested. However, only M. javanica-infected 'Petite Yellow' and 'Maui' had decreases (P 24%, whereas 'Tifway' appeared to be relatively tolerant with only a 4% reduction in root dry weight. Twenty other Cynodon accessions showed decreases (P 11% root reductions. In addition to 'Tifway,' 10 other Cynodon accessions and L. perenne, F. arundinacea, D. decumbens, and S. bicolor appeared to be relatively more tolerant of B. longicaudatus than the other accessions evaluated. PMID- 19283058 TI - Sampling approaches for extensive surveys in nematology. AB - Extensive surveys of the frequency and abundance of plant-parasitic nematodes over large geographic areas provide useful data of unknown reliability. Time, cost, and logistical constraints may limit the sampling intensity that can be invested at any survey site. We developed a computer program to evaluate the probability of detection and the reliability of population estimates obtained by different strategies for collecting one sample of 10 cores from a field. We used data from two fields that had been sampled systematically and extensively as the basis for our analyses. Our analyses indicate that, at least for those two fields, it is possible to have a high probability of detecting the presence of nematode species and to reliably estimate abundance, with a single 10-core soil sample from a field. When species were rare or not uniformly distributed in a field, the probability of detection and reliability of the population estimate were correlated with the distance between core removal sites. Increasing the prescribed distance between cores resulted in the composite sample representing a wider range of microenvironments in the field. PMID- 19283059 TI - Occurrence of Meloidogyne spp. in Argentina. AB - A record of 84 plant species in 32 families that are hosts to the root-knot nematode species found in Argentina is presented. The genus Meloidogyne appears to be widely distributed in the country, with Meloidogyne incognita and M. javanica the most frequently detected species. Other species found in Argentina include M. arenaria, M. cruciani, M. decalineata, M. hapla, and M. ottersoni. The present survey is supplemented with existing published information. PMID- 19283060 TI - Survey of nematodes on coffee in hawaii. AB - Surveys of coffee fields in Hawaii during 1989-1991 indicated the presence of 10 nematode species in 8 genera. After coffee was planted in fields previously in sugarcane, populations of Criconemella sp. and Pratylenchus zeae gradually decreased, while Rotylenchulus reniformis and, in one field, Meloidogyne incognita, increased in numbers. Coffee is a poor host of R. reniformis, but weeds in coffee plantations may support this nematode. At present, nematodes pose no serious threat to Hawaii's expanding coffee industry. PMID- 19283061 TI - Distribution of Heterodera carotae and Meloidogyne hapla in Michigan Carrot Production. AB - During 1986 and 1988, selected farms in all of the major carrot-growing counties of Michigan were surveyed to determine the extent of infestation by Heterodera carotae and Meloidogyne hapla. Both species were found in all eight counties surveyed, but not on all farms. Heterodera carotae was recovered from 67.4% of the fields surveyed. Meloidogyne hapla was detected in 24.8% of the samples and from 69.8% of the fields. In most cases, H. carotae and M. hapla occurred in the same field. PMID- 19283062 TI - Survey of Plant-parasitic Nematodes in Missouri Cotton Fields. AB - During September 1990, 30 cotton fields in each of three Missouri counties were surveyed for plant-parasitic nematodes. Soil samples for nematode analysis consisted of a composite of 20 cores collected in a zig-zag pattern within a 1-ha block in each field. Cores were taken from within weed-free cotton rows. Nine genera of plant-parasitic nematodes were found (Rotylenchulus, Helicotylenchus, Hoplolaimus, Meloidogyne, Paratylenchus, Pratylenchus, Tylenchorhynchus, Heterodera, and Trichodorus), and five species were identified: Meloidogyne incognita, Rotylenchulus reniformis, Hoplolaimus galeatus, Pratylenchus vulnus, and P. scribneri. This is the first report of R. reniformis, H. galeatus, P. vulnus, and P. scribneri in Missouri cotton fields and the first report of R. reniformis and P. vulnus in Missouri. The known cotton pathogens M. incognita, R. reniformis, and H. galeatus were found in 30%, 3%, and 2% of the fields sampled, respectively. The correlation between sand content of the soil sample and the number of vermiform M. incognita in the sample was not significant, with r(2) = 0.13. Select fields where H. galeatus and R. reniformis were found in 1990 were sampled more intensely in 1991. The 1-ha block sampled in 1990 was sampled in 1991, along with three other 1-ha blocks uniformly distributed within the field. In addition, a 1-ha block was sampled in each of eight nearby fields, within 2 km of the first field. The nine plant-parasitic nematode genera identified in the 1990 survey were observed again in 1991. Within-field distribution of M. incognita, R. reniformis, and H. galeatus was not uniform. When M. incognita, R. reniformis, or H. galeatus were present in a field, the same species was found in 38%, 25%, or 50% of nearby fields, respectively. PMID- 19283063 TI - The complete genome and proteome of Laribacter hongkongensis reveal potential mechanisms for adaptations to different temperatures and habitats. AB - Laribacter hongkongensis is a newly discovered Gram-negative bacillus of the Neisseriaceae family associated with freshwater fish-borne gastroenteritis and traveler's diarrhea. The complete genome sequence of L. hongkongensis HLHK9, recovered from an immunocompetent patient with severe gastroenteritis, consists of a 3,169-kb chromosome with G+C content of 62.35%. Genome analysis reveals different mechanisms potentially important for its adaptation to diverse habitats of human and freshwater fish intestines and freshwater environments. The gene contents support its phenotypic properties and suggest that amino acids and fatty acids can be used as carbon sources. The extensive variety of transporters, including multidrug efflux and heavy metal transporters as well as genes involved in chemotaxis, may enable L. hongkongensis to survive in different environmental niches. Genes encoding urease, bile salts efflux pump, adhesin, catalase, superoxide dismutase, and other putative virulence factors-such as hemolysins, RTX toxins, patatin-like proteins, phospholipase A1, and collagenases-are present. Proteomes of L. hongkongensis HLHK9 cultured at 37 degrees C (human body temperature) and 20 degrees C (freshwater habitat temperature) showed differential gene expression, including two homologous copies of argB, argB-20, and argB-37, which encode two isoenzymes of N-acetyl-L-glutamate kinase (NAGK) NAGK-20 and NAGK-37-in the arginine biosynthesis pathway. NAGK-20 showed higher expression at 20 degrees C, whereas NAGK-37 showed higher expression at 37 degrees C. NAGK-20 also had a lower optimal temperature for enzymatic activities and was inhibited by arginine probably as negative-feedback control. Similar duplicated copies of argB are also observed in bacteria from hot springs such as Thermus thermophilus, Deinococcus geothermalis, Deinococcus radiodurans, and Roseiflexus castenholzii, suggesting that similar mechanisms for temperature adaptation may be employed by other bacteria. Genome and proteome analysis of L. hongkongensis revealed novel mechanisms for adaptations to survival at different temperatures and habitats. PMID- 19283064 TI - Sequential assembly of centromeric proteins in male mouse meiosis. AB - The assembly of the mitotic centromere has been extensively studied in recent years, revealing the sequence and regulation of protein loading to this chromosome domain. However, few studies have analyzed centromere assembly during mammalian meiosis. This study specifically targets this approach on mouse spermatocytes. We have found that during prophase I, the proteins of the chromosomal passenger complex Borealin, INCENP, and Aurora-B load sequentially to the inner centromere before Shugoshin 2 and MCAK. The last proteins to be assembled are the outer kinetochore proteins BubR1 and CENP-E. All these proteins are not detected at the centromere during anaphase/telophase I and are then reloaded during interkinesis. The loading sequence of the analyzed proteins is similar during prophase I and interkinesis. These findings demonstrate that the interkinesis stage, regularly overlooked, is essential for centromere and kinetochore maturation and reorganization previous to the second meiotic division. We also demonstrate that Shugoshin 2 is necessary for the loading of MCAK at the inner centromere, but is dispensable for the loading of the outer kinetochore proteins BubR1 and CENP-E. PMID- 19283065 TI - Recombinational landscape and population genomics of Caenorhabditis elegans. AB - Recombination rate and linkage disequilibrium, the latter a function of population genomic processes, are the critical parameters for mapping by linkage and association, and their patterns in Caenorhabditis elegans are poorly understood. We performed high-density SNP genotyping on a large panel of recombinant inbred advanced intercross lines (RIAILs) of C. elegans to characterize the landscape of recombination and, on a panel of wild strains, to characterize population genomic patterns. We confirmed that C. elegans autosomes exhibit discrete domains of nearly constant recombination rate, and we show, for the first time, that the pattern holds for the X chromosome as well. The terminal domains of each chromosome, spanning about 7% of the genome, exhibit effectively no recombination. The RIAILs exhibit a 5.3-fold expansion of the genetic map. With median marker spacing of 61 kb, they are a powerful resource for mapping quantitative trait loci in C. elegans. Among 125 wild isolates, we identified only 41 distinct haplotypes. The patterns of genotypic similarity suggest that some presumed wild strains are laboratory contaminants. The Hawaiian strain, CB4856, exhibits genetic isolation from the remainder of the global population, whose members exhibit ample evidence of intercrossing and recombining. The population effective recombination rate, estimated from the pattern of linkage disequilibrium, is correlated with the estimated meiotic recombination rate, but its magnitude implies that the effective rate of outcrossing is extremely low, corroborating reports of selection against recombinant genotypes. Despite the low population, effective recombination rate and extensive linkage disequilibrium among chromosomes, which are techniques that account for background levels of genomic similarity, permit association mapping in wild C. elegans strains. PMID- 19283066 TI - An Epstein-Barr virus anti-apoptotic protein constitutively expressed in transformed cells and implicated in burkitt lymphomagenesis: the Wp/BHRF1 link. AB - Two factors contribute to Burkitt lymphoma (BL) pathogenesis, a chromosomal translocation leading to c-myc oncogene deregulation and infection with Epstein Barr virus (EBV). Although the virus has B cell growth-transforming ability, this may not relate to its role in BL since many of the transforming proteins are not expressed in the tumor. Mounting evidence supports an alternative role, whereby EBV counteracts the high apoptotic sensitivity inherent to the c-myc-driven growth program. In that regard, a subset of BLs carry virus mutants in a novel form of latent infection that provides unusually strong resistance to apoptosis. Uniquely, these virus mutants use Wp (a viral promoter normally activated early in B cell transformation) and express a broader-than-usual range of latent antigens. Here, using an inducible system to express the candidate antigens, we show that this marked apoptosis resistance is mediated not by one of the extended range of EBNAs seen in Wp-restricted latency but by Wp-driven expression of the viral bcl2 homologue, BHRF1, a protein usually associated with the virus lytic cycle. Interestingly, this Wp/BHRF1 connection is not confined to Wp-restricted BLs but appears integral to normal B cell transformation by EBV. We find that the BHRF1 gene expression recently reported in newly infected B cells is temporally linked to Wp activation and the presence of W/BHRF1-spliced transcripts. Furthermore, just as Wp activity is never completely eclipsed in in vitro transformed lines, low-level BHRF1 transcripts remain detectable in these cells long-term. Most importantly, recognition by BHRF1-specific T cells confirms that such lines continue to express the protein independently of any lytic cycle entry. This work therefore provides the first evidence that BHRF1, the EBV bcl2 homologue, is constitutively expressed as a latent protein in growth-transformed cells in vitro and, in the context of Wp-restricted BL, may contribute to virus associated lymphomagenesis in vivo. PMID- 19283067 TI - Abnormal brain iron homeostasis in human and animal prion disorders. AB - Neurotoxicity in all prion disorders is believed to result from the accumulation of PrP-scrapie (PrP(Sc)), a beta-sheet rich isoform of a normal cell-surface glycoprotein, the prion protein (PrP(C)). Limited reports suggest imbalance of brain iron homeostasis as a significant associated cause of neurotoxicity in prion-infected cell and mouse models. However, systematic studies on the generality of this phenomenon and the underlying mechanism(s) leading to iron dyshomeostasis in diseased brains are lacking. In this report, we demonstrate that prion disease-affected human, hamster, and mouse brains show increased total and redox-active Fe (II) iron, and a paradoxical increase in major iron uptake proteins transferrin (Tf) and transferrin receptor (TfR) at the end stage of disease. Furthermore, examination of scrapie-inoculated hamster brains at different timepoints following infection shows increased levels of Tf with time, suggesting increasing iron deficiency with disease progression. Sporadic Creutzfeldt-Jakob disease (sCJD)-affected human brains show a similar increase in total iron and a direct correlation between PrP and Tf levels, implicating PrP(Sc) as the underlying cause of iron deficiency. Increased binding of Tf to the cerebellar Purkinje cell neurons of sCJD brains further indicates upregulation of TfR and a phenotype of neuronal iron deficiency in diseased brains despite increased iron levels. The likely cause of this phenotype is sequestration of iron in brain ferritin that becomes detergent-insoluble in PrP(Sc)-infected cell lines and sCJD brain homogenates. These results suggest that sequestration of iron in PrP(Sc)-ferritin complexes induces a state of iron bio-insufficiency in prion disease-affected brains, resulting in increased uptake and a state of iron dyshomeostasis. An additional unexpected observation is the resistance of Tf to digestion by proteinase-K, providing a reliable marker for iron levels in postmortem human brains. These data implicate redox-iron in prion disease-associated neurotoxicity, a novel observation with significant implications for prion disease pathogenesis. PMID- 19283068 TI - Timing cellular decision making under noise via cell-cell communication. AB - Many cellular processes require decision making mechanisms, which must act reliably even in the unavoidable presence of substantial amounts of noise. However, the multistable genetic switches that underlie most decision-making processes are dominated by fluctuations that can induce random jumps between alternative cellular states. Here we show, via theoretical modeling of a population of noise-driven bistable genetic switches, that reliable timing of decision-making processes can be accomplished for large enough population sizes, as long as cells are globally coupled by chemical means. In the light of these results, we conjecture that cell proliferation, in the presence of cell-cell communication, could provide a mechanism for reliable decision making in the presence of noise, by triggering cellular transitions only when the whole cell population reaches a certain size. In other words, the summation performed by the cell population would average out the noise and reduce its detrimental impact. PMID- 19283069 TI - Identification and gene expression analysis of a taxonomically restricted cysteine-rich protein family in reef-building corals. AB - The amount of genomic sequence information continues to grow at an exponential rate, while the identification and characterization of genes without known homologs remains a major challenge. For non-model organisms with limited resources for manipulative studies, high-throughput transcriptomic data combined with bioinformatics methods provide a powerful approach to obtain initial insights into the function of unknown genes. In this study, we report the identification and characterization of a novel family of putatively secreted, small, cysteine-rich proteins herein named Small Cysteine-Rich Proteins (SCRiPs). Their discovery in expressed sequence tag (EST) libraries from the coral Montastraea faveolata required the performance of an iterative search strategy based on BLAST and Hidden-Markov-Model algorithms. While a discernible homolog could neither be identified in the genome of the sea anemone Nematostella vectensis, nor in a large EST dataset from the symbiotic sea anemone Aiptasia pallida, we identified SCRiP sequences in multiple scleractinian coral species. Therefore, we postulate that this gene family is an example of lineage-specific gene expansion in reef-building corals. Previously published gene expression microarray data suggest that a sub-group of SCRiPs is highly responsive to thermal stress. Furthermore, data from microarray experiments investigating developmental gene expression in the coral Acropora millepora suggest that different SCRiPs may play distinct roles in the development of corals. The function of these proteins remains to be elucidated, but our results from in silico, transcriptomic, and phylogenetic analyses provide initial insights into the evolution of SCRiPs, a novel, taxonomically restricted gene family that may be responsible for a lineage-specific trait in scleractinian corals. PMID- 19283070 TI - Cage matching: head to head competition experiments of an invasive plant species from different regions as a means to test for differentiation. AB - Many hypotheses are prevalent in the literature predicting why some plant species can become invasive. However, in some respects, we lack a standard approach to compare the breadth of various studies and differentiate between alternative explanations. Furthermore, most of these hypotheses rely on 'changes in density' of an introduced species to infer invasiveness. Here, we propose a simple method to screen invasive plant species for potential differences in density effects between novel regions. Studies of plant competition using density series are a fundamental tool applied to virtually every aspect of plant population ecology to better understand evolution. Hence, we use a simple density series with substitution contrasting the performance of Centaurea solstitialis in monoculture (from one region) to mixtures (seeds from two regions). All else being equal, if there is no difference between the introduced species in the two novel regions compared, Argentina and California, then there should be no competitive differences between intra and inter-regional competition series. Using a replicated regression design, seeds of each species were sown in the greenhouse at 5 densities in monoculture and mixed and grown till onset of flowering. Centaurea seeds from California had higher germination while seedlings had significantly greater survival than Argentina. There was no evidence for density dependence in any measure for the California region but negative density dependence was detected in the germination of seeds from Argentina. The relative differences in competition also differed between regions with no evidence of differential competitive effects of seeds from Argentina in mixture versus monoculture while seeds from California expressed a relative cost in germination and relative growth rate in mixtures with Argentina. In the former instance, lack of difference does not mean 'no ecological differences' but does suggest that local adaptation in competitive abilities has not occurred. Importantly, this method successfully detected differences in the response of an invasive species to changes in density between novel regions which suggests that it is a useful preliminary means to explore invasiveness. PMID- 19283071 TI - The Werner syndrome helicase/exonuclease processes mobile D-loops through branch migration and degradation. AB - RecQ DNA helicases are critical for preserving genome integrity. Of the five RecQ family members identified in humans, only the Werner syndrome protein (WRN) possesses exonuclease activity. Loss of WRN causes the progeroid disorder Werner syndrome which is marked by cancer predisposition. Cellular evidence indicates that WRN disrupts potentially deleterious intermediates in homologous recombination (HR) that arise in genomic and telomeric regions during DNA replication and repair. Precisely how the WRN biochemical activities process these structures is unknown, especially since the DNA unwinding activity is poorly processive. We generated biologically relevant mobile D-loops which mimic the initial DNA strand invasion step in HR to investigate whether WRN biochemical activities can disrupt this joint molecule. We show that WRN helicase alone can promote branch migration through an 84 base pair duplex region to completely displace the invading strand from the D-loop. However, substrate processing is altered in the presence of the WRN exonuclease activity which degrades the invading strand both prior to and after release from the D-loop. Furthermore, telomeric D-loops are more refractory to disruption by WRN, which has implications for tighter regulation of D-loop processing at telomeres. Finally, we show that WRN can recognize and initiate branch migration from both the 5' and 3' ends of the invading strand in the D-loops. These findings led us to propose a novel model for WRN D-loop disruption. Our biochemical results offer an explanation for the cellular studies that indicate both WRN activities function in processing HR intermediates. PMID- 19283072 TI - Pre-Columbian origins for North American anthrax. AB - Disease introduction into the New World during colonial expansion is well documented and had a major impact on indigenous populations; however, few diseases have been associated with early human migrations into North America. During the late Pleistocene epoch, Asia and North America were joined by the Beringian Steppe ecosystem which allowed animals and humans to freely cross what would become a water barrier in the Holocene. Anthrax has clearly been shown to be dispersed by human commerce and trade in animal products contaminated with Bacillus anthracis spores. Humans appear to have brought B. anthracis to this area from Asia and then moved it further south as an ice-free corridor opened in central Canada approximately 13,000 ybp. In this study, we have defined the evolutionary history of Western North American (WNA) anthrax using 2,850 single nucleotide polymorphisms (SNPs) and 285 geographically diverse B. anthracis isolates. Phylogeography of the major WNA B. anthracis clone reveals ancestral populations in northern Canada with progressively derived populations to the south; the most recent ancestor of this clonal lineage is in Eurasia. Our phylogeographic patterns are consistent with B. anthracis arriving with humans via the Bering Land Bridge. This northern-origin hypothesis is highly consistent with our phylogeographic patterns and rates of SNP accumulation observed in current day B. anthracis isolates. Continent-wide dispersal of WNA B. anthracis likely required movement by later European colonizers, but the continent's first inhabitants may have seeded the initial North American populations. PMID- 19283073 TI - A computational approach to understand in vitro alveolar morphogenesis. AB - Primary human alveolar type II (AT II) epithelial cells maintained in Matrigel cultures form alveolar-like cysts (ALCs) using a cytogenesis mechanism that is different from that of other studied epithelial cell types: neither proliferation nor death is involved. During ALC formation, AT II cells engage simultaneously in fundamentally different, but not fully characterized activities. Mechanisms enabling these activities and the roles they play during different process stages are virtually unknown. Identifying, characterizing, and understanding the activities and mechanisms are essential to achieving deeper insight into this fundamental feature of morphogenesis. That deeper insight is needed to answer important questions. When and how does an AT cell choose to switch from one activity to another? Why does it choose one action rather than another? We report obtaining plausible answers using a rigorous, multi-attribute modeling and simulation approach that leveraged earlier efforts by using new, agent and object oriented capabilities. We discovered a set of cell-level operating principles that enabled in silico cells to self-organize and generate systemic cystogenesis phenomena that are quantitatively indistinguishable from those observed in vitro. Success required that the cell components be quasi-autonomous. As simulation time advances, each in silico cell autonomously updates its environment information to reclassify its condition. It then uses the axiomatic operating principles to execute just one action for each possible condition. The quasi-autonomous actions of individual in silico cells were sufficient for developing stable cyst-like structures. The results strengthen in silico to in vitro mappings at three levels: mechanisms, behaviors, and operating principles, thereby achieving a degree of validation and enabling answering the questions posed. We suggest that the in silico operating principles presented may have a biological counterpart and that a semiquantitative mapping exists between in silico causal events and in vitro causal events. PMID- 19283074 TI - Discovery of novel hypermethylated genes in prostate cancer using genomic CpG island microarrays. AB - BACKGROUND: Promoter and 5' end methylation regulation of tumour suppressor genes is a common feature of many cancers. Such occurrences often lead to the silencing of these key genes and thus they may contribute to the development of cancer, including prostate cancer. METHODOLOGY/PRINCIPAL FINDINGS: In order to identify methylation changes in prostate cancer, we performed a genome-wide analysis of DNA methylation using Agilent human CpG island arrays. Using computational and gene-specific validation approaches we have identified a large number of potential epigenetic biomarkers of prostate cancer. Further validation of candidate genes on a separate cohort of low and high grade prostate cancers by quantitative MethyLight analysis has allowed us to confirm DNA hypermethylation of HOXD3 and BMP7, two genes that may play a role in the development of high grade tumours. We also show that promoter hypermethylation is responsible for downregulated expression of these genes in the DU-145 PCa cell line. CONCLUSIONS/SIGNIFICANCE: This study identifies novel epigenetic biomarkers of prostate cancer and prostate cancer progression, and provides a global assessment of DNA methylation in prostate cancer. PMID- 19283075 TI - A GRFa2/Prop1/stem (GPS) cell niche in the pituitary. AB - BACKGROUND: The adult endocrine pituitary is known to host several hormone producing cells regulating major physiological processes during life. Some candidates to progenitor/stem cells have been proposed. However, not much is known about pituitary cell renewal throughout life and its homeostatic regulation during specific physiological changes, such as puberty or pregnancy, or in pathological conditions such as tumor development. PRINCIPAL FINDINGS: We have identified in rodents and humans a niche of non-endocrine cells characterized by the expression of GFRa2, a Ret co-receptor for Neurturin. These cells also express b-Catenin and E-cadherin in an oriented manner suggesting a planar polarity organization for the niche. In addition, cells in the niche uniquely express the pituitary-specific transcription factor Prop1, as well as known progenitor/stem markers such as Sox2, Sox9 and Oct4. Half of these GPS (GFRa2/Prop1/Stem) cells express S-100 whereas surrounding elongated cells in contact with GPS cells express Vimentin. GFRa2+-cells form non-endocrine spheroids in culture. These spheroids can be differentiated to hormone-producing cells or neurons outlining the neuroectoderm potential of these progenitors. In vivo, GPSs cells display slow proliferation after birth, retain BrdU label and show long telomeres in its nuclei, indicating progenitor/stem cell properties in vivo. SIGNIFICANCE: Our results suggest the presence in the adult pituitary of a specific niche of cells characterized by the expression of GFRa2, the pituitary specific protein Prop1 and stem cell markers. These GPS cells are able to produce different hormone-producing and neuron-like cells and they may therefore contribute to postnatal pituitary homeostasis. Indeed, the relative abundance of GPS numbers is altered in Cdk4-deficient mice, a model of hypopituitarism induced by the lack of this cyclin-dependent kinase. Thus, GPS cells may display functional relevance in the physiological expansion of the pituitary gland throughout life as well as protection from pituitary disease. PMID- 19283076 TI - Has the data efflux regarding the promising outcome following injection of deflux changed the management of adult vesicoureteral reflux? AB - Primary vesicoureteral reflux (VUR), traditionally considered a problem of childhood, can also be detected during adulthood. However, while the concept regarding the therapeutic management of VUR in children has undergone revolutionary changes, moving from surgical to conservative approach, the optimal therapeutic approach in adult reflux is poorly addressed and is still unknown. Herein, we review clinical and therapeutic approaches of VUR in pediatric population as published throughout the years. With the introduction of Deflux injection as a minimally invasive procedure, we identify a beginning of a new trend that further extends the indications for endoscopic injections, including its introduction to adult patients as well. PMID- 19283077 TI - The changing concepts of vesicoureteral reflux in children. PMID- 19283078 TI - Characterization of arterial wave reflection in healthy bonnet macaques: feasibility of applanation tonometry. AB - Nonhuman primates are commonly used in cardiovascular research. Increased arterial stiffness is a marker of subclinical atherosclerosis and higher CV risk. We determined the augmentation index (AI) using applanation tonometry in 61 healthy monkeys (59% female, age 1-25 years). Technically adequate studies were obtained in all subjects and required 1.5 +/- 1.3 minutes. The brachial artery provided the highest yield (95%). AI was correlated with heart rate (HR) (r = 0.65, P < .001), crown rump length (CRL) (r = 0.42, P = .001), and left ventricular (LV) mass determined using echocardiography (r = 0.52, P < .001). On multivariate analysis, HR (P < .001) and CRL (P = .005) were independent predictors of AI (R2 = 0.46, P < .001). Body Mass Index (BMI) and AI were independent predictors of higher LV mass on multivariate analysis (P < .001 and P = .03). In conclusion, applanation tonometry is feasible for determining AI. Reference values are provided for AI in bonnet macaques, in whom higher AI is related to HR and CRL, and in turn contributes to higher LV mass. PMID- 19283079 TI - High-throughput detection of induced mutations and natural variation using KeyPoint technology. AB - Reverse genetics approaches rely on the detection of sequence alterations in target genes to identify allelic variants among mutant or natural populations. Current (pre-) screening methods such as TILLING and EcoTILLING are based on the detection of single base mismatches in heteroduplexes using endonucleases such as CEL 1. However, there are drawbacks in the use of endonucleases due to their relatively poor cleavage efficiency and exonuclease activity. Moreover, pre screening methods do not reveal information about the nature of sequence changes and their possible impact on gene function. We present KeyPoint technology, a high-throughput mutation/polymorphism discovery technique based on massive parallel sequencing of target genes amplified from mutant or natural populations. KeyPoint combines multi-dimensional pooling of large numbers of individual DNA samples and the use of sample identification tags ("sample barcoding") with next generation sequencing technology. We show the power of KeyPoint by identifying two mutants in the tomato eIF4E gene based on screening more than 3000 M2 families in a single GS FLX sequencing run, and discovery of six haplotypes of tomato eIF4E gene by re-sequencing three amplicons in a subset of 92 tomato lines from the EU-SOL core collection. We propose KeyPoint technology as a broadly applicable amplicon sequencing approach to screen mutant populations or germplasm collections for identification of (novel) allelic variation in a high-throughput fashion. PMID- 19283080 TI - Abstract Rule Learning for Visual Sequences in 8- and 11-Month-Olds. AB - The experiments reported here investigated the development of a fundamental component of cognition: to recognize and generalize abstract relations. Infants were presented with simple rule-governed patterned sequences of visual shapes (ABB, AAB, and ABA) that could be discriminated from differences in the position of the repeated element (late, early, or nonadjacent, respectively). Eight-month olds were found to distinguish patterns on the basis of the repetition, but appeared insensitive to its position in the sequence; 11-month-olds distinguished patterns over the position of the repetition, but appeared insensitive to the nonadjacent repetition. These results suggest that abstract pattern detection may develop incrementally in a process of constructing complex relations from more primitive components. PMID- 19283081 TI - PPARgamma in Kidney Physiology and Pathophysiology. AB - Involvement of the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) in kidney physiology has been explored recently. Synthetic PPARgamma ligands can ameliorate the diabetic kidney disease through different mechanisms, involving inhibition of mesangial cell growth, reduction of mesangial matrix, and cytokine production of glomerular cells as well as promoting endothelial cell survival within the kidney glomeruli. Activation of PPARgamma has additional profibrotic consequences, which can contribute to wound healing in diabetic glomerulonephritis. Beside many beneficial effects, PPARgamma activation, however, can lead to severe water retention, a common side effect of thiazolidinedione therapy. This unwanted effect is due to the activation of PPARgamma in the mesonephric distal collecting system, where PPARgamma positively regulates sodium and water resorbtion leading to the expansion of interstitial fluid volume. Recent studies indicate that PPARgamma is also involved in the normal kidney development, renal lipid metabolism, and activation of the renin angiotensin system. In this paper, we give a synopsis of the current knowledge on PPARgamma functions in kidney phyisology and pathophysiology. PMID- 19283082 TI - Panic disorder: is the PAG involved? AB - Data from studies with humans have suggested that abnormalities of midbrain structures, including the periaqueductal gray matter (PAG), could be involved in the neurobiology of panic disorder (PD). The electrical stimulation of the PAG in neurosurgical patients induces panic-like symptoms and the effect of drugs that are effective in the treatment of PD in the simulation of public speaking model of anxiety is in agreement with data from animal models of PD. Structural neuroimaging studies have shown increases in gray matter volume of midbrain and pons of PD patients. There is also evidence of lower serotonin transporter and receptor binding, and increases of metabolism in the midbrain of PD patients. Nevertheless, these midbrain abnormalities can not be considered as specific findings, since neuroimaging data indicate that PD patients have abnormalities in other brain structures that process fear and anxiety. PMID- 19283083 TI - Controversies in the management of borderline resectable proximal pancreatic adenocarcinoma with vascular involvement. AB - Synchronous major vessel resection during pancreaticoduodenectomy (PD) for borderline resectable pancreatic adenocarcinoma remains controversial. In the 1970s, regional pancreatectomy advocated by Fortner was associated with unacceptably high morbidity and mortality rates, with no impact on long-term survival. With the establishment of a multidisciplinary approach, improvements in preoperative staging techniques, surgical expertise, and perioperative care reduced mortality rates and improved 5-year-survival rates are now achieved following resection in high-volume centres. Perioperative morbidity and mortality following PD with portal vein resection are comparable to standard PD, with reported 5-year-survival rates of up to 17%. Segmental resection and reconstruction of the common hepatic artery/proper hepatic artery (CHA/PHA) can be performed to achieve an R0 resection in selected patients with limited involvement of the CHA/PHA at the origin of the gastroduodenal artery (GDA). PD with concomitant major vessel resection for borderline resectable tumours should be performed when a margin-negative resection is anticipated at high-volume centres with expertise in complex pancreatic surgery. Where an incomplete (R1 or R2) resection is likely neoadjuvant treatment with systemic chemotherapy followed by chemoradiation as part of a clinical trial should be offered to all patients. PMID- 19283084 TI - Cholesterol-dependent anaplasma phagocytophilum exploits the low-density lipoprotein uptake pathway. AB - In eukaryotes, intracellular cholesterol homeostasis and trafficking are tightly regulated. Certain bacteria, such as Anaplasma phagocytophilum, also require cholesterol; it is unknown, however, how this cholesterol-dependent obligatory intracellular bacterium of granulocytes interacts with the host cell cholesterol regulatory pathway to acquire cholesterol. Here, we report that total host cell cholesterol increased >2-fold during A. phagocytophilum infection in a human promyelocytic leukemia cell line. Cellular free cholesterol was enriched in A. phagocytophilum inclusions as detected by filipin staining. We determined that A. phagocytophilum requires cholesterol derived from low-density lipoprotein (LDL), because its replication was significantly inhibited by depleting the growth medium of cholesterol-containing lipoproteins, by blocking LDL uptake with a monoclonal antibody against LDL receptor (LDLR), or by treating the host cells with inhibitors that block LDL-derived cholesterol egress from late endosomes or lysosomes. However, de novo cholesterol biosynthesis is not required, since inhibition of the biosynthesis pathway did not inhibit A. phagocytophilum infection. The uptake of fluorescence-labeled LDL was enhanced in infected cells, and LDLR expression was up-regulated at both the mRNA and protein levels. A. phagocytophilum infection stabilized LDLR mRNA through the 3' UTR region, but not through activation of the sterol regulatory element binding proteins. Extracellular signal-regulated kinase (ERK) was up-regulated by A. phagocytophilum infection, and inhibition of its upstream kinase, MEK, by a specific inhibitor or siRNA knockdown, reduced A. phagocytophilum infection. Up regulation of LDLR mRNA by A. phagocytophilum was also inhibited by the MEK inhibitor; however, it was unclear whether ERK activation is required for LDLR mRNA up-regulation by A. phagocytophilum. These data reveal that A. phagocytophilum exploits the host LDL uptake pathway and LDLR mRNA regulatory system to accumulate cholesterol in inclusions to facilitate its replication. PMID- 19283085 TI - Biophysically realistic filament bending dynamics in agent-based biological simulation. AB - An appealing tool for study of the complex biological behaviors that can emerge from networks of simple molecular interactions is an agent-based, computational simulation that explicitly tracks small-scale local interactions--following thousands to millions of states through time. For many critical cell processes (e.g. cytokinetic furrow specification, nuclear centration, cytokinesis), the flexible nature of cytoskeletal filaments is likely to be critical. Any computer model that hopes to explain the complex emergent behaviors in these processes therefore needs to encode filament flexibility in a realistic manner. Here I present a numerically convenient and biophysically realistic method for modeling cytoskeletal filament flexibility in silico. Each cytoskeletal filament is represented by a series of rigid segments linked end-to-end in series with a variable attachment point for the translational elastic element. This connection scheme allows an empirically tuning, for a wide range of segment sizes, viscosities, and time-steps, that endows any filament species with the experimentally observed (or theoretically expected) static force deflection, relaxation time-constant, and thermal writhing motions. I additionally employ a unique pair of elastic elements--one representing the axial and the other the bending rigidity- that formulate the restoring force in terms of single time-step constraint resolution. This method is highly local -adjacent rigid segments of a filament only interact with one another through constraint forces-and is thus well-suited to simulations in which arbitrary additional forces (e.g. those representing interactions of a filament with other bodies or cross-links / entanglements between filaments) may be present. Implementation in code is straightforward; Java source code is available at www.celldynamics.org. PMID- 19283087 TI - Influence of Selected Plant Species on Hatching of Eggs and Development of Juveniles of Heterodera glycines. AB - The influence of selected plant species on egg hatch and subsequent development of Heterodera glycines race 3 was investigated. Plants tested included four soybean cultivars, red clover, alfalfa, hairy vetch, field corn, sweet corn, cabbage, tobacco, cotton, and wheat. Soybean stimulated egg hatching more than any of the other plant species, with H. glycines-resistant cultivars being more stimulating than susceptible ones. Hairy vetch also increased hatch. Roots of cabbage, red clover, alfalfa, and hairy vetch were readily penetrated by juveniles of H. glycines. Maturation to adult occurred only on soybean and hairy vetch. PMID- 19283088 TI - Pathogenicity of Steinernema scapterisci to Selected Invertebrates. AB - Steinernema scapterisci was more pathogenic to insects tested in the order Orthoptera than to those in the orders Lepidoptera or Hymenoptera; it was not pathogenic to earthworms. The nematode also infected and killed the mole crickets Scapteriscus acletus and S. vicinus when released four successive times at 10-day intervals in containers of soil infested with the nematode. PMID- 19283086 TI - Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process. AB - A key process in the lifecycle of the malaria parasite Plasmodium falciparum is the fast invasion of human erythrocytes. Entry into the host cell requires the apical membrane antigen 1 (AMA-1), a type I transmembrane protein located in the micronemes of the merozoite. Although AMA-1 is evolving into the leading blood stage malaria vaccine candidate, its precise role in invasion is still unclear. We investigate AMA-1 function using live video microscopy in the absence and presence of an AMA-1 inhibitory peptide. This data reveals a crucial function of AMA-1 during the primary contact period upstream of the entry process at around the time of moving junction formation. We generate a Plasmodium falciparum cell line that expresses a functional GFP-tagged AMA-1. This allows the visualization of the dynamics of AMA-1 in live parasites. We functionally validate the ectopically expressed AMA-1 by establishing a complementation assay based on strain-specific inhibition. This method provides the basis for the functional analysis of essential genes that are refractory to any genetic manipulation. Using the complementation assay, we show that the cytoplasmic domain of AMA-1 is not required for correct trafficking and surface translocation but is essential for AMA-1 function. Although this function can be mimicked by the highly conserved cytoplasmic domains of P. vivax and P. berghei, the exchange with the heterologous domain of the microneme protein EBA-175 or the rhoptry protein Rh2b leads to a loss of function. We identify several residues in the cytoplasmic tail that are essential for AMA-1 function. We validate this data using additional transgenic parasite lines expressing AMA-1 mutants with TY1 epitopes. We show that the cytoplasmic domain of AMA-1 is phosphorylated. Mutational analysis suggests an important role for the phosphorylation in the invasion process, which might translate into novel therapeutic strategies. PMID- 19283089 TI - Reproductive and Damage Potential of Ditylenchus destructor on Peanut. AB - The reproductive and damage potential of Ditylenchus destructor on peanut, Arachis hypogaea cv. Sellie, was determined in greenhouse tests. Final nematode population densities (Pf) in roots, hulls, and seeds increased (P = 0.01) as a function of increasing initial population (Pi). Final population densities were higher in hulls than in seeds and roots. Final densities in hulls and seeds were positively (P = 0.01) correlated. Fresh root and hull weight and number of pods and seeds per plant were not affected by D. destructor. Second generation germination and pod and seed disease severity increased (P = 0.01), whereas fresh seed weight decreased (P = 0.01) as a function of increasing Pi, and Pf in seeds and Pf in hulls. At Pi 250 and higher, 10-25% of seeds germinated into second generation seedlings before harvest. At Pi 250 and higher, fresh weight of harvested seed was suppressed 20-50%. At Pi 50 or Pf greater than 20 per seed, pod disease severity was 3-7 (on a scale of 1 to 10) and 15-80% of seeds were blemished or unsound. PMID- 19283090 TI - Effects of Aldicarb and Fenamiphos on Acetycholinesterase and Motility of Caenorhabditis elegans. AB - The ability of Caenorhabditis elegans to recover from exposure to high doses of aldicarb and fenamiphos was examined at the organismal and biochemical levels by determination of movement and acetylcholinesterase activity. Nematodes recovered rapidly from a 24-hour exposure to both compounds at concentrations that caused complete paralysis. Acetylcholinesterase regained nearly full activity after a 24 hour exposure to aldicarb but only 10% activity after exposure to fenamiphos. The nematodes were able to move normally, however, on the limited activity that was regained after fenamiphos treatment. Mutant C. elegans strains deficient in various molecular forms of acetylcholinesterase were utilized to demonstrate that the mechanism of recovery did not involve new synthesis of enzyme. This result was confirmed by experiments on acetylcholinesterase reactivation from live versus dead nematodes. PMID- 19283091 TI - Carbon Dioxide and Temperature Gradielits in Baermann Funnel Extraction of Rotylenchulus reniformis. AB - Vermiform Rotylenchulus reniformis were anesthetized in water by 10-40% CO but were fully motile for 24 hours in water below 5% CO. When air containing 2.5% CO was blown onto agar, nematodes accumulated at the point of highest CO concentration. Nematodes also accumulated when chilling (0.2-1 C) of agar by the gas flow at the accumulation point was offset with heat from a fiber optic. In Baermann funnels containing R. reniformis in silt loam and sandy clay loam soils, CO in funnel water increased during 24 hours from 0 to ca. 1%; more CO accumulated below the soil layer than above. Bubbling air with 2.5% CO into water below soil in covered funnels increased the CO gradient and increased nematode extraction, whereas bubbling air without CO below soil purged CO from the water and decreased nematode extraction. Manipulation of CO within funnels usually increased extraction by only 30% and never by more than 3-fold. Controlling temperature gradients consistently increased extraction by 2-30-fold. PMID- 19283092 TI - Observations on Crystalloid Bodies in the Pseudocoelom of Eutobrilus heptapapillatus. AB - Eutobrilus heptapapillatus, found in a number of different sites at the mouth of the River Murray in South Australia, were examined under light and electron microscopes. The pseudocoeloms of these nematodes often contained oval crystalloid bodies containing carbohydrate, sulfur, phosphorus, and lipid. The bodies varied considerably in size up to a maximum of 10 mum long. The precise function of these crytalloids remains unknown. Nematodes having these crystalloids often also contained numerous small regular densely staining particles, about 20 nm d and occurring throughout the nematode's body. PMID- 19283093 TI - Mesocriconema ornicauda n. sp. and Ogma floridense n. sp. (Nematoda: Criconematidae) from Two Florida Habitats. AB - Mesocriconema ornicauda n. sp. and Ogma floridense n. sp. are described from two native habitats of central and northwestern Florida. Mesocriconema ornicauda is closest to M. annulatiforme (De Grisse &Loof, 1967) Loof &De Grisse, 1989, but differs by the shorter stylet of the female (43-50 vs. 54-65 mum) and the moderately conoid tail of the male, which is pointed in M. annulatiforme. Ogma floridense is closest to O. hungaricum (Andrassy, 1972) Siddiqi, 1986. Females differ from those of O. hungaricum by the first of two labial annuli being wider, whereas they are subequal in O. hungaricum. Ogma floridense females differ also by entire or bifid cuticular scales, which are consistently divided into two or four projections in O. hungaricum, the shorter body (360-471 vs. 480-550 mum), the shorter stylet (87-98 vs. 95-100 mum), and the more anteriorly located excretory pore (Rex = 17-19 vs. 21-23). PMID- 19283094 TI - Population Development of Pasteuria penetrans on Meloidogyne arenaria. AB - A microplot study on the influence of cropping sequences with peanut in summer and bare fallowed or cover crops of rye or vetch in winter on the population development of Pasteuria penetrans was initiated in the spring of 1987. The number of spores of P. penetrans attached per second-stage juvenile of Meloidogyne arenaria race 1 increased from 0.11 in the fall of 1987 to 7.6, 8.6, and 3.6 in the fall of 1989 in the rye, vetch, and fallowed plots, respectively. Higher (P /= 0.05) root growth at any temperature or Pi. The UT3 nematode reproductive indices were greater than those of the other nematode populations at all Pi and increased with temperature. PMID- 19283143 TI - Degree-day Models for Predicting Egg Hatch and Population Increase of Criconemella xenoplax. AB - A degree-day model was derived to predict egg hatch for Criconemella xenoplax. Eggs collected from gravid females were incubated in distilled water at constant temperatures of 10-35 C. Sixty-six percent of all eggs hatched between 13 and 32 C, and 42% hatched at 10 C. All eggs aborted above 32.5 C. Between 25 and 32 C, 8.5 +/- 0.5 days were required for egg hatch. Degree-day requirement for egg hatch at 10-30 C was estimated to be 154 +/- 5 with a base of 9.03 +/- 0.04 C. This base of 9 C was adopted in studies of the relationship between degree-days and nematode population increase on Prunus seedlings grown 9-11 weeks in a greenhouse. Degree-day accumulations were based upon daily averages from maximum and minimum air temperatures. Ratios of final to initial population densities exhibited an exponential pattern in relation to degree-day accumulations with proportionate doubling increment of 0.100 +/- 0.049 every 139 +/- 8 degree-days. These results provide a means of predicting nematode population increase under greenhouse conditions and a basis for choosing sampling intervals when evaluating nematode multiplication. PMID- 19283144 TI - Evaluation of Host Suitability in Prunus for Criconemella xenoplax. AB - Methods were developed for screening Prunus selections for host suitability to Criconemella xenoplax. The relative host suitability of selections was based upon a doubling accumulation value (beta) that was defined as the number of degree days (base 9 C) required for doubling of an increment of the initial nematode population. The beta value characteristic for C. xenoplax (139 +/- 8 degree-days) on suitable hosts was similar to the average beta value determined for several peach rootstocks known to be suitable hosts. The beta values were 144 +/- 21 for Halford, 141 +/- 16 for Lovell, and 138 +/- 10 for Nemaguard. A higher value for beta could indicate poorer host suitability or resistance of a selection to C. xenoplax. All of 369 Prunus accessions tested, including eight accessions that had survived well on a field site infested with C. xenoplax, were suitable hosts. Apparently, resistance to C. xenoplax was not a factor in survival of the accessions planted in the field. Seedlings from P. besseyi, P. pumila 'Mando', and two interspecific hybrids, Redcoat and Sapalta IR 549-1, failed to support nematode population increase in 44-81% of tests conducted, but all selections supported population increase in some tests. These accessions may have resistance mechanisms that are active only under specific conditions. PMID- 19283145 TI - Benomyl Tolerance of Ten Fungi Antagonistic to Plant-parasitic Nematodes. AB - Ten strains of fungi were tested for tolerance to the fungicide benomyl. Verticillium chlamydosporium strain 2 did not grow in the presence of benomyl; Drechraeria coniospora strains 1 and 2 and Chaetomium sp. tolerated only 0.1 mug benomyl/ml medium; Acremonium bacillisporum, an unidentified fungus, and Phoma chrysanthemicola uniformly grew at 1 mug/ml, but some hyphae grew at higher benomyl concentrations; Fusarium sp. tolerated 475 mug/ml, but some hyphae grew on medium amended with 1,000 mug/ml; Verticillium lecanii and V. chlamydosporium strain 1 routinely tolerated 1,000 mug/ml. Fungi generally grew more slowly at higher than at lower benomyl concentrations. Strains with elevated tolerance to benomyl were selected from Acremonium bacillisporum, Drechmeria coniospora, Fusarium sp., and an unidentified fungus. These strains retained the increased tolerance after repeated transfers on unamended medium. PMID- 19283146 TI - Plant protection with inorganic ions. AB - Gradients of salts of the specific ion repellents for Meloidogyne incognita -- NH, K, Cl, and NO -- have been demonstrated to shield tomato roots from infestation in soil. The strategy of these greenhouse experiments was to interpose a salt barrier in a soil column between the plant roots and the nematodes. The relative effectiveness of the salts as a barrier to infective second-stage juveniles in a sandy loam was NHNO, NHCl > KNO > KCl. Some of these ions are beneficial to plant growth, and the results suggest that a new environmentally tolerable means of plant protection is possible. PMID- 19283147 TI - Characterization of species and races of the genus meloidogyne by DNA restriction enzyme analysis. AB - Total DNA of three species of Meloidogyne spp., including four subspecific races of M. incognita, were digested separately with EcoR I, Cla III, and Hind III and probed with (3)(2)P-labelled total genomic DNA from M. incognita race 1 in Southern hybridizations. Short exposures of Southern blots after Hind III digestion revealed patterns that were useful for separating the species. Race differences were seen after longer exposures. The DNA fragment patterns obtained were scanned with a laser densitometer and the data were subjected to principal coordinate and cluster analyses. The likelihood of cloning species and race specific DNA probes is discussed. PMID- 19283148 TI - Evidence Against Amplification of Four Genes in Giant Cells Induced by Meloidogyne incognita. AB - Giant-cell DNA was isolated from pea (Pisum sativum) inoculated with Meloidogyne incognita and used in slot blots to test for selective sequence amplification. Four sequences representing low (ribulose 1,5-bisphosphate carboxylase and actin), mid-level (histone 3), and highly repetitive (large ribosomal repeat) sequence DNA were used as probes. Known amounts of root-tip DNA and giant-cell DNA were blotted onto hybridization membranes and probed. The signal strength on autoradiographs containing equal amounts of root-tip DNA and giant-cell DNA were compared with a scanning densitometer. No difference in signal strength between equal amounts of root-tip DNA and giant-cell DNA was found. Thus, for the probes tested, there is no difference in copy number and, hence, no selective DNA sequence amplification has occurred. PMID- 19283149 TI - Abscisic Acid and Ethylene Increase in Heterodera avenae-infected Tolerant or Intolerant Oat Cultivars. AB - The relationship between root stunting caused by the cereal cyst nematode and levels of two root growth inhibiting hormones, abscisic acid and ethylene, was investigated in aseptically cultured root segments and in intact roots of two oat cultivars differing in tolerance to the nematode. Cultured root segments of oat cultivars New Zealand Cape (tolerant) and Sual (intolerant) were inoculated with sterilized Heterodera avenae second-stage juveniles. Suppressed growth of root axes and emerged laterals following nematode penetration corresponded to an increase in abscisic acid and ethylene in roots of both intolerant and tolerant cultivars. When the experiment was repeated on intact root systems, nematodes retarded root growth of Sual more than New Zealand Cape despite an increase in ABA and ethylene in both cultivars. Abscisic acid and (or) ethylene may be involved in growth inhibition of H. avenae-infected roots but appear to play no direct role in determining tolerance. PMID- 19283150 TI - Cryopreservation of Steinernema carpocapsae and Heterorhabditis bacteriophora. AB - A method for the cryopreservation of third-stage infective juveniles (IJ) of Steinernema carpocapsae and Heterorhabiditis bacteriophora was developed. Cryoprotection was achieved by incubating the nematodes in 22% glycerol (S. carpocapsae) or 14% glycerol (H. bacteriophora) for 24 hours, followed by 70% methanol at 0 C for 10 minutes. The viability of S. carpocapsae frozen in liquid nitrogen as 20 mul volumes spread over cover slip glass was > 80%. Survival of H. bacteriophora frozen on glass varied from 10 to 60% but was improved to > 80% by replacing the glass with filter paper. Cryopreservation and storage of 1-ml aliqots of S. carpocapsae IJ resulted in > 50% survival after 8 months; pathogenicity was retained and normal in vitro development took place. Trehalose and glycerol levels increased and glycogen levels decreased during incubation of S. carpocapsae IJ in glycerol. Normal levels of trehalose, glycerol and glycogen were restored during post freezing rehydration. PMID- 19283151 TI - Cryopreservation of the Pinewood Nematode, Bursaphelenchus spp. AB - Populations of three isolates of Bursaphelenchus xylophilus, the pinewood nematode, and one of B. mucronatus were treated with three cryoprotectants at -70 C for 24 hours followed by deep freezing at -180 C in liquid nitrogen for different periods of time. A solution of 15% glycerol, 35% buffer S, and 50% M9, or 1% aqueous solution of dimethylsulfoxide (DMSO), or a mixture of 60% M9 and 40% S buffer were used as cryoprotectants. A significantly larger number of juveniles than adults survived deep freezing. Significantly more nematodes were motile after cryopreservation in the 15% glycerol-S-M9 soludon than in the M9-S buffer solution or the DMSO aqueous solution. When cryopreserved nematodes that had been treated with glycerol solution were plated onto Botrytis cinerea, they reproduced rapidly over several generations. Cryopreserved nematodes were as pathogenic as untreated nematodes to Scots pines. PMID- 19283152 TI - Infection of Cultured Thin Cell Layer Roots of Lycopersicon esculentum by Meloidogyne incognita. AB - A new aseptic culture system for studying interactions between tomato (Lycopersicon esculentum) and Meloidogyne incognita is described. Epidermal thin cell layer explants from peduncles of tomato produced up to 20 adventitious roots per culture in 4-9 days on Murashige &Scoog medium plus kinetin and indole acetic acid. Rooted cultures were transferred to Gamborg's B-5 medium and inoculated with infective second-stage juveniles. Gall formation was apparent 5 days after inoculation and egg production by mature females occurred within 25 days at 25 C in the susceptible genotypes Rutgers and Red Alert. Resistant genotypes LA655, LA656, and LA1022 exhibited a characteristic hypersensitive response. This system provides large numbers of cultured root tips for studies on the molecular basis of the host-parasite relationship. PMID- 19283153 TI - A bioassay to estimate root penetration by nematodes. AB - An in vitro bioassay with a 96-well microtiter plate was used to study the effect of lectins on burrowing nematode penetration of citrus roots. In each well, one 4 mm root segment, excised from the zone of elongation of rough lemon roots, was buried in 0.88 g dry sand. Addition of a Radopholus citrophilus suspension containing ca. 300 nematodes in 50 mu1 test solution completely moistened the sand in each well. The technique assured uniform treatment concentration throughout the medium. Within 16-24 hours, burrowing nematodes penetrated citrus root pieces, primarily through the cut ends. The lectins (100 mug/ml) Concanavalin A (Con A), soybean agglutinin (SBA), wheat germ agglutinin (WGA), and Lotus tetragonolobus agglutinin (LOT) stimulated an increase in penetration of citrus root segments by Radopholus citrophilus. Concentrations as low as 12.5 mug/ml Con A, LOT, and WGA stimulated burrowing nematode penetration of citrus roots. Heat denaturation of the lectins reversed their effect on penetration; however, incubation of nematodes in lectin (25 mug/ml) with 25 mM competitive sugars did not. The reason for enhanced penetration associated with lectins is unclear. PMID- 19283154 TI - Wheat Germ Agglutinin Bound to the Outer Cuticle of the Seed Gall Nematodes Anguina agrostis and A. tritici. AB - The presence of wheat germ agglutinin (WGA) on the cuticular surface of the seed gall nematodes Anguina agrostis and Anguina tritici was demonstrated, and the nature of its binding was examined. Crude extracts from the cuticles of A. tritici agglutinated human red blood cells, and only N-acetylglucosamine (GlucNAc) inhibited the agglutination. Distribution of the lectin was visualized by treating live infective juveniles (J2) with rabbit anti-WGA antibody and staining with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG. The lectin bound to the outer cuticular surface of the whole body wall. Pretreatment with GlucNAc oligomers did not reduce the fluorescence created by the anti-WGA-WGA binding, indicating at least a partial nonspeciflc adhesion of the WGA to the nematode surface. Proteolytic enzyme pretreatments diminished the fluorescence, whereas lipase and periodate pretreatments increased the fluorescence. Adult females and males were labeled only on the head and tail, whereas eggs were not labeled at all. It was concluded that the WGA on the J2 cuticle originates from the host. PMID- 19283155 TI - Comparisons of Isozyme Phenotypes in Five Meloidogyne spp. with Isoelectric Focusing. AB - Meloidogyne incognita race 1, M. javanica, M. arenaria race 1, M. hapla, and an undescribed Meloidogyne sp. were analyzed by comparing isozyme phenotypes of esterase, malate dehydrogenase, phosphoglucomutase, isocitrate dehydrogenase, and alpha-glycerophosphate dehydrogenase. Isozyme phenotypes were obtained from single mature females by isoelectric focusing electrophoresis. Of these five isozymes, only esterase and phosphoglucomutase could be used to separate all five Meloidogyne spp.; however, the single esterase electromorphs were similar for M. incognita and M. hapla. Yet when both nematodes were run on the same gel, differences in their esterase phenotypes were detectable. Isozyme phenotypes from the other three isozymes revealed a great deal of similarity among M. incognita, M. javanica, M. arenaria, and the undescribed Meloidogyne sp. PMID- 19283156 TI - Interaction of Meloidogyne incognita and Water Stress in Two Cotton Cultivars. AB - A series of controlled-environment experiments were conducted to elucidate the effects of Meloidogyne incognita on host physiology and plant-water relations of two cotton (Gossypium hirsutum) cultivars that differed in their susceptibility to nematode infection. Inoculation of M. incognita-resistant cultivar Auburn 634 did not affect growth, stomatal resistance, or components of plant-water potential relative to uninoculated controls. However, nematode infection of the susceptible cultivar Stoneville 506 greatly suppressed water flow through intact roots. This inhibition exceeded 28% on a root-length basis and was similar to that observed as a consequence of severe water stress in a high evaporative demand environment. Nematodes did not affect the components of leaf water potential, stomatal resistance, transpiration, or leaf temperature. However, these factors were affected by the interaction of M. incognita and water stress. Our results indicate that M. incognita infection may alter host-plant water balance and may be a significant factor in early-season stress on cotton seedlings. PMID- 19283157 TI - Development of Meloidogyne arenaria on Peanut and Soybean under Two Temperature Cycles. AB - Florunner peanut and three soybean cultivars, Centennial, Gasoy 17, and Wright, were inoculated with 48-hour age cohorts of Meloidogyne arenari race 1 second stage juveniles and placed in a growth chamber set to simulate early season (low temperature) and midseason (high temperature) conditions. Percentages of the initial inoculum penetrating roots 4 and 8 days after inoculation were 2-3 times higher in soybean cultivars than in peanut; 25% on susceptible soybean and 9% on peanut. Penetration and early development of M. arenaria were greater in the higher temperature environment. Penetration percentages were expressed as a function of cumulative degree-days by regression models. Development of M. arenaria 10, 20, and 30 days after inoculation was more rapid on peanut than on soybean. The resistant soybean cultivar Wright had slower development rates than did the other two soybean cultivars. Nematode growth and development were dependent on temperature. In greenhouse experiments, production of eggs by M. arenaria was more than 10 times greater on peanut than on susceptible soybean. The reproductive factor for Wright soybean was less than one, but plant growth parameters indicated that this cultivar was intolerant of M. arenavia. PMID- 19283158 TI - Incubation Temperature and Time Effects on Life Stages of Bursaphelenchus xylophilus in Wood Chips. AB - Wood chips of Pinus strobus inoculated with Bursaphelenchus xylophilus were incubated at 3, 12, 30, or 40 C during intervals of 47, 82, and 130 days to determine the effects of incubation temperature and time on total number of nematodes and occurrence of each life stage. Nematodes did not survive at 40 C; the greatest number of nematodes was maintained at 3 C. The number and percentage of juveniles in the propagative cycle were greatest at 3 C after 47 days, but the percentage was greatest at 30 C after 130 days. More third-stage dispersal larvae, with percentages as high as 85%, were extracted at 3 and 12 C than at 30 C by the end of the study. Dauer larvae were extracted from the chips, but percentages never exceeded 5%. The percentage of adults was greater at 30 C than at 3 and 12 C after 82 and 130 days. When a 1-week heat treatment at 30 C was applied to samples at 3 and 12 C,. numbers and percentages of adults increased. Percentages of dauer larvae increased very slightly when the heat treatment was applied after 47 days, but numbers and percentages of juveniles and dispersals were affected erratically. PMID- 19283159 TI - Population Dynamics of Ditylenchus destructor on Peanut. AB - The population development of Ditylenchus destructor in the roots, pegs, hulls, and seeds of eight peanut (Arachis hypogaea) genotypes was studied in the greenhouse. Although all genotypes tested were good hosts for D. destructor, differences in host suitability were observed. Invasion of the plant parts by Ditylenchus destructor proceeded more slowly in genotypes with long growth periods. During the second half of the growth period of these genotypes, D. destructor populations emigrated from the hulls and seeds into the soil but reinfected the pods after a few weeks. The genotypes with the longest growth periods supported the highest number of nematodes when each genotype was harvested at its usual harvest time. The long-season genotypes supported low numbers of nematodes when harvested before crop maturity. PMID- 19283160 TI - The Genus Prochromadora with a Redescription of P. orleji from a Marine Saltern in the People's Republic of China. AB - The genus Prochromadora Filipjev, 1922 comprising nine species is reviewed and a key is presented. Salient diagnostic characters of the genus are a single, large, solid dorsal tooth and a cuticle showing transverse rows of homogenous punctations that extend around the body without intervening lateral differentiation. A redescription and drawings of P. orleji from a solar saltworks in Qingdao, Shandong, the People's Republic of China, are presented. PMID- 19283161 TI - Taxonomic notes on some hemicriconemoides species and description of a new species. AB - Hemicriconemoides scottolamassesei n. sp., named after Dr. C. Scotto la Massese, from New Caledonia, is described and illustrated. Primary diagnostic characters are females with a continuous head and three lip annules, the first annule being the smallest, stylet length of 65-75 mum, vulva without lateral flaps, and acute tail terminus; J4 has a smooth cuticle without scales or spines; the male has a lateral field with two incisures. Emended measurements, descriptions, illustrations, and new records of distribution are given for H. brachyurus (Loos, 1949) Chitwood &Birchfield, 1957, H. mangiferae Siddiqi, 1961, and H. kanayaensis Nakasono &Ichinoe, 1961. Relationships of H. promissus Vovlas, 1980, based on study of paratypes, and H. intermedius Dasgupta, Raski &Van Gundy, 1969 are discussed; both are proposed as synonyms of H. brachyurus. PMID- 19283163 TI - Comparison of treatment means: a statistical fantasy. PMID- 19283162 TI - Extraction Efficiency of Belonolaimus longicaudatus from Sandy Soil. AB - Numbers of Belonolaimus longicaudatus extracted from sandy soils (91-92% sand) by sieving and centrifugation were only 40-55% of those extracted by sieving and incubation on a Baermann tray. Residues normally discarded at each step of the sieving plus Baermann tray extraction procedure were examined for nematodes to obtain estimates of extraction efficiencies. For third-stage and fourth-stage juveniles, males, and females, estimates of extraction efficiency ranged from 60 to 65% in one experiment and 73 to 82% in another. Estimated extraction efficiencies for second-stage juveniles were lower (33% in one experiment, 67% in another) due to losses during sieving. When sterilized soil was seeded with known numbers of B. longicaudatus, 60% of second-stage juveniles and 68-76% of other stages were recovered. Most stages of B. longicaudatus could be extracted from these soils by sieving plus Baermann incubation with an efficiency of 60-70%. PMID- 19283164 TI - Reaction of Prunus Rootstocks to Meloidogyne incognita and M. arenaria in Spain. AB - Prunus rootstocks were evaluated for their reaction to Meloidogyne incognita and M. arenaria. Most rootstocks were peach-almond hybrids of Spanish origin. In one experiment three selections of Garfi x Nemared (G x N) and Hansen-5 were highly resistant to M. incognita, but four other rootstocks were susceptible showing high galling indices and population increases. In two experiments with M. arenaria, the hybrid selections G x N nos. 1 and 9 were immune, GF-305 and Hansen 5 were resistant, but nine other rootstocks expressed various degrees of susceptibility. All Spanish rootstocks were susceptible to both Meloidogyne species except for the three G x N selections. The root-knot nematode resistant peach Nemared used as a male parent with Garfi was found to transmit a high degree of resistance to M. incognita and immunity to M. arenaria. Progenies of P. davidiana (Ga x D no. 3), a known source of resistance to root-knot nematodes, were susceptible. PMID- 19283165 TI - Host Suitability of Eight Prunus spp. and One Pyrus communis Rootstocks to Pratylenchus vulnus, P. neglectus, and P. thornei. AB - The effects of Pratylenchus vulnus on rootstocks of eight commonly used Prunus spp. and one Pyrus communis were evaluated under greenhouse conditions during a 15-month period. In a first experiment, two almonds (Moncayo and Garrigues), one peach (GF-305), and two peach-almond hybrids (GF-677 and Adafuel) inoculated with 2,000 nematodes per plant proved to be good hosts of P. vulnus. Highest (P < 0.05) numbers of nematodes per gram of fresh root weight were recovered from Adafuel and GF-677. Root weights were higher in uninoculated compared to inoculated plants of all rootstocks, whereas top weights of uninoculated Garrigues, GF-305, and GF-677 differed (P < 0.05) from those of inoculated plants. In a second experiment, three plum (Marianna 2624, Myrobalan 605, and San Julian 655-2) and one pear (OHF-333) rootstocks were also found to be good hosts of P. vulnus, although significantly fewer nematodes were recovered from Myrohalan 605 roots than from the other three materials. Inoculated OHF-333 and San Julian 655-2 differed (P < 0.05) in root weights over uninoculated plants. Only inoculated San Julian 655-2 showed differences in top weights over uninoculated treatments. Rootstocks were poor or non-hosts for P. neglectus and P. thornei. PMID- 19283166 TI - Resistance in Soybean Cultivars from Maturity Groups V-VIII to Soybean Cyst and Root-knot Nematodes. AB - One hundred thirty-nine cultivars of soybean (Glycine max) in Maturity Groups V, VI, VII, and VIII were evaluated in a greenhouse for resistance to Heterodera glycines races 3 and 14 and Meloidogyne incognita, M. arenaria, and M. javanica. Of the cultivars tested, 37% had resistance to H. glycines race 3 alone, 12% had resistance to both races 3 and 14, and 69% exhibited a moderate or high level of resistance to one or more of the Meloidogyne spp. However, 24% were susceptible to each race of H. glycines and to all Meloidogyne spp. Whereas 40% of the cultivars were moderately resistant to M. javanica, only 23% were moderately resistant to M. incognita and 24% to M. arenaria. Although 28% of the cultivars had a high level of resistance to M. incognita, only 6 and 3% had this level of resistance to M. javanica and M. arenaria, respectively. Seventeen percent of the cultivars possessed a moderate or high level of resistance to all three Meloidogyne spp., and 37 % had resistance to H. glycines race 3 and M. incognita. With the exception of resistance to H. glycines race 14, resistance in soybean to these nematodes was fairly uniformly distributed across maturity groups. PMID- 19283167 TI - Soybean Cultivars Resistant and Susceptible to Heterodera glycines. AB - Additional tests of 178 cultivars and lines of soybean against soybean cyst nematode are reported. Cultivars are available with some level of resistance to races 1, 2, 3, 4, 5, 6, 9, and 14, but in some cases the resistance level is moderate. No cultivar or line (except possibly PI 437654) has resistance to all of the races tested. Thirteen cultivars and lines, however, were resistant or moderately resistant to race 1, 14 to race 2, 87 to race 3, 2 to race 4, 3 to race 5, 42 to race 6, 18 to race 9, and 36 to race 14. Some have resistance to more than one race. PMID- 19283168 TI - Reaction of Trifolium repens Cultivars and Germplasms to Meloidogyne incognita. AB - Ten cultivars and 13 germplasms of white clover (Trifolium repens) were evaluated in the greenhouse for resistance to the southern root-knot nematode, Meloidogyne incognita race 4. One hundred plants of each cultivar or germplasm were rated for percentage of the root system galled (PRSG) at 60 days after inoculation with root-knot nematode eggs. Tillman (9%) and SRVR (19%) had the highest percentage of resistant plants (PRSG = 0 or 1 on a scale of 0-5 ) for the cultivars and germplasms, respectively. No resistant plants were selected from the cultivars California Ladino or Sacramento, or from the germplasms Brown Loam population or Brown Loam Synthetic #6. Resistant plants identified in this study were used to initiate are current selection program for resistance to M. incognita. PMID- 19283169 TI - Response of Tomato Cultigens to Meloidogyne javanica and Races of Meloidogyne incognita. AB - Thirty-six cultigens of tomato were evaluated for resistance against Meloidogyne javanica and four races of M. incognita with standards and parameters adopted by the International Meloidogyne Project. Most cultigens were susceptible to the nematodes, including some that were previously reported to be resistant to these nematodes. Ten accessions, namely Pusa-120, Calmart VFN, Panjab 6.NR-7, EC173898 (72T6), EC173897 (Cal-Mart), EC173896 (Kewalo), CLN363BCF-167-1-0, CLN363BCF-190 1-0, CLN363BCF-344-0-0, and CLN299BCF-4-1-4-1-1-0, were immune to all test nematodes. VFN-Bush and VFN-8 were resistant to all four races of M. incognita and immune to M. javanica. Three cultivars (Pant-T, Money Maker, and Pelican) exhibited a degree of race-specific resistance to M. incognita. Pant-T and Money Maker were hypersusceptible to race 1 and race 4 of M. incognita, respectively, but were susceptible to other races. Pelican was tolerant to M. incognita race 3 but resistant to the other races. PMID- 19283170 TI - Susceptibility of Diploid St. Augustinegrasses to Belonolaimus longicaudatus. AB - A fine-textured, dwarf St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) genotype, FX-313, was severely damaged in plots in the third year of evaluation in sandy soil in southern Florida. Damage was associated with numerous (> 40/100-cm(3) soil) sting nematodes, Belonolaimus longicaudatus Rau. Damage was ameliorated (P < 0.05) by fenamiphos applied broadcast at 2.2 g a.i./m(2), and B. longicaudatus numbers were reduced (P < 0.01), compared with untreated plots. Root dry weights of four diploid (2n = 18) St. Augustinegrasses--FX-261, FX-299, FX-313, and Seville--were reduced (P < 0.001) by B. longicaudatus in a temperature- and light-controlled experiment. Estimated daily transpiration, an indicator of plant health, was reduced (P < 0.001) after 112 days to 3.32 g/pot for inoculated plants, compared with 5.10 g /pot for uninoculated plants. Genotypes did not differ in nematode number per pot (mean 551/215 cm(2) soil) 128 days after inoculation, but differed (P < 0.05) in nematode numbers on a root dry weight basis, with FX-313 and Seville representing the extremes, 12,300 and 4,000 B. longicaudatus/g root dry weight, respectively. The diploid St. Augustinegrasses evaluated were good hosts for B. longicaudatus, but field data and controlled inoculation demonstrate genetic variation in susceptibility. PMID- 19283171 TI - Resistance to Root-knot Nematodes in Cucumber and Horned Cucumber. AB - Two experiments were conducted in the greenhouse. In one experiment, cucumber (Cucumis sativus) and horned cucumber (C. metuliferus) cultigens were evaluated for resistance to four root-knot nematode species (Meloidogyne arenaria, M. hapla, M. incognita, and M. javanica), and, in a second experiment, a standard (12-week) test was compared with a rapid (6-week) test. In the first experiment, horned cucumber cultigens varied in response to the Meloidogyne species. 'Sumter' cucumber was more susceptible than the horned cucumber to Meloidogyne incognita, M. javanica, and M. arenaria. All cultigens were more resistant to M. hapla than to the other root-knot nematode species. In the second experiment, best results were obtained when the test was run for 12 weeks rather than 6 weeks after planting (or 10 and 4 weeks after inoculation, respectively). All cultigens were more resistant to M. arenaria than to either M. incognita or M. javanica. PMID- 19283172 TI - Race Composition of Meloidogyne incognita and M. arenaria Populations in Vegetable Fields in Uttar Pradesh. AB - A total of 1,256 populations of Meloidogyne incognita and 442 populations of M. arenaria were collected from vegetables in eight districts of Uttar Pradesh, India. Host differentials were used to identify the host race of each population. All four host races of M. incognita were present in six of the eight districts. In the other two districts, only host races 1, 2, and 4 were found. Although frequencies of occurrence of the races differed among districts, races 1 and 2 comprised 62% of all M. incognita populations evaluated. Only host race 2 of M. arenaria was found in this survey, and this race was found in all eight districts. This is the first report of M. arenaria host race 2 from India. PMID- 19283173 TI - Nematodes associated with blackberry in arkansas. AB - A survey of the nematodes in blackberry (Rubus sp.) rhizospheres was conducted in Arkansas from 1986 to 1989. The state was divided arbitrarily into four quadrants. A total of 134 soil samples was collected, and 150-cm 3 subsamples were assayed for nematodes. Twenty-one species of plant-parasitic nematodes in 11 genera were extracted from the samples. There were differences (P = 0.05) among quadrants of the state in percentage occurrence of the nematodes and in population densities in samples. Xiphinema americanum, Helicotylenchus spp. (H. paraplatyurus, H. platyurus, and H. pseudorobustus), and Pratylenchus spp. (P. vulnus and P. zeae) were found in all quadrants. Xiphinema americanum population density was near 1,000 per 150 cm(3) soil in soil samples from two locations. Other nematodes found in one or more quadrants were Criconemella spp. (C. axeste, C. curvata, C. denoudeni, C. ornata, C. sphaerocephala, and C. xenoplax), Paratrichodorus minor, Tylenchorhynchus claytoni, Hirschmanniella oryzae, Hoplolaimus magnistylus, Scutellonema bradys, and undescribed species of Criconema, Tylenchulus, Xiphinema, and Meloidogyne. Criconemella sphaerocephala and Helicotylenchus platyurus are reported from Arkansas for the first time. Helicotylenchus paraplatyurus is reported from the United States for the first time. PMID- 19283174 TI - Distribution of Heterodera glycines Races in Illinois. AB - Because the race scheme for Heterodera glycines was expanded recently from 5 to 16 races, the occurrence and distribution of races in Illinois need clarification. Forty-four populations of H. glycines were collected from sites in 23 of the 88 infested counties. Populations were tested using the differential soybean lines Pickett 71, Peking, PI 88788, and PI 90763. Lee 68 and Williams 82 were used as standards. Seedlings were grown in 7.5-cm-d clay pots and inoculated with 1,000 eggs and second-stage juveniles obtained from H. glycines-infested field soil. Plants were maintained in a greenhouse at 22-28 C. After 1 month, the number of first-generation white females that developed on each differential was determined and the race of the population was designated. Twenty-eight populations were race 3, twelve were race 1, two were race 5, one was race 2, and one was race 4. Populations of races 3 and 1 were widely distributed in the state. In 26 of the 28 race determinations, race designations using Williams 82 and Lee 68 were the same, indicating that if Lee 68 is not available, Williams 82 may be a suitable alternative for race tests done in the north central United States. PMID- 19283175 TI - Nematodes associated with dieback disease of cranberries. AB - Hemicycliophora ritteri and Paratrichodorus minor were associated in high numbers with a previously unknown "dieback disease" of cranberry. Chemical treatment with nematicides and fungicides failed to control this disease. There were no correlations (P 0.99) to concentrations between 0 and 10,000 nematodes/ml. Coefficients of variation ranged from 12-23%, depending on the species and developmental stage used. The method, therefore, was at least as accurate as direct counts of nematodes in aliquots on a microscope and more than 100 times as fast. The method should have direct application in research programs on plant resistance to nematodes, nematode population dynamics, and nematode behavior. PMID- 19283208 TI - Population densities of five migratory endoparasitic nematodes in carrot disk cultures. AB - Numbers of nematodes recovered per culture varied greatly among five species cultured on carrot disks. Radopholus similis and Pratylenchus vulnus showed the highest population densities, with 23,400-fold and 16,600-fold increases, respectively, in 90 days. Final populations of P. thornei and Zygotytenchus guevarai were similar but lower than those of R. similis and P. vulnus. The population of P. neglectus increased 74 times. Species with the greatest reproduction in this study reproduce sexually. PMID- 19283209 TI - Modeling of Yield Loss in Potato Early Dying Caused by Pratylenchus penetrans and Verticillium dahliae. AB - Yield-loss models were developed for potato early dying, caused by an interaction between Verticillium dahliae and Pratylenchus penetrans. Yield data were collected over 5 years (1985-1989) from potato plants grown in microplots infested with V. dahliae and (or) P. penetrans. The model y = b + (1 - b)/(1 + [VD/36.7]), where y was the relative yield (with uninfested controls = 1.0) and VD was the preplant density of V. dahliae microsclerotia per cm(3) soil, was fitted to the data set. When P. penetrans = 0, b = 0.55 (SE = 0.099), and when P. penetrans > 0, b = 0.23 (SE = 0.035). This model assumed that yield loss was proportional to the concentration of preplant microsclerotia of V. dahliae, and only qualitatively related to presence or absence of P. penetrans. This study contrasts with previous reports that predict yield loss being proportional to preplant population densities of both P. penetrans and V. dahliae. PMID- 19283210 TI - Leaching Soluble Salts Increases Population Densities of Tylenchulus semipenetrans. AB - The effect of salinity on population densities of Tylenchulus semipenetrans was measured on 3-month-old salt-tolerant Rangpur lime growing on either loamy sand, sand, or organic mix and on 4-month-old salt-sensitive Sweet lime in organic mix. Salinity treatments were initiated by watering daily with 25 mol/m(3) NaCl + 3.3 mol/m(3) CaCl for 3 days and every other day with 50 mol/m(3) NaC1 + 6.6 mol/m(3) CaC1 for one week, with no salt (NS) treatments as controls. Salinity was discontinued in one treatment (DS) by leaching with tap water prior to inoculation with nematodes, whereas the continuous salinity (CS) treatment remained unchanged. Overall, in Rangpur lime organic soil supported the highest population densities of T. semipenetrans, followed by loamy sand and sand. The DS treatment resulted in the highest (P /= 1,111 J2/100 cm(3) soil) in all inoculated plots. Pod yield and dry weight of foliage at harvest were negatively correlated (P 65 years, owning home BP monitor, numbers enrolling and numbers attending 12 month follow up. RESULTS: Of 524 eligible patients, 414 (79%) enrolled in the clinic, of whom 89% completed the trial. At 12 months, HBP control rates rose from 29.9% to 44.8%, with mean HBP falling 5.2/3.2 mmHg (p<0.001). Home BP monitor ownership rose from 54.3 to 82.9%. DISCUSSION: This is the first study in standard Australian general practice using both a comprehensive clinic approach and HBP readings exclusively. This study provided a feasible management protocol and practical clinical performance indicators that could be used for a randomised controlled trial. Significantly better control rates were achieved compared with published studies for BP control. PMID- 19283259 TI - Paediatric vascular birthmarks--the psychological impact and the role of the GP. AB - BACKGROUND: This study was designed to investigate the psychosocial impact on the family of a child with a vascular birthmark and examine the role of the general practitioner in meeting the family's needs. METHOD: Nineteen families were interviewed with a questionnaire before their assessment at the Sydney Children's Hospital (New South Wales) Vascular Birthmarks Clinic. RESULTS: Sixty-eight percent of parents overestimated the size of their child's lesion when asked to draw it; 15% said some lifestyle modifications had to be made to accommodate their child's condition; 63% were concerned about their child being teased at school; and 36% had issues of self blame and embarrassment. Expectations from the clinic included information (68%), treatment (47%) and reassurance (26%). CONCLUSION: Vascular birthmarks are common. Treatment strategies are improving, but there is a need to adequately address the psychosocial impact that these lesions have, both on parents and children. The GP is the carer best placed to meet these needs. PMID- 19283260 TI - Fear of needles--nature and prevalence in general practice. AB - BACKGROUND: The extent to which a fear of needles influences health decisions remains largely unexplored. This study investigated the prevalence of fear of needles in a southeast Queensland community, described associated symptoms, and highlighted health care avoidance tendencies of affected individuals. METHODS: One hundred and seventy-seven participants attending an outer urban general practice responded to a questionnaire on fear of needles, symptoms associated with needles and its influence on their use of medical care. RESULTS: Twenty-two percent of participants reported a fear of needles. Affected participants were more likely than participants with no fear to report vasovagal symptoms, have had a previous traumatic needle experience (46.2 vs. 16.4%, p<0.001) and avoid medical treatment involving needles (20.5 vs. 2.3%, p<0.001). DISCUSSION: Fear of needles is common and is associated with health care avoidance. Health professionals could better identify and manage patients who have a fear of needles by recognising associated vasovagal symptoms and past traumatic experiences. PMID- 19283261 TI - Screened hybrid density functionals for solid-state chemistry and physics. AB - Density functional theory incorporating hybrid exchange-correlation functionals has been extraordinarily successful in providing accurate, computationally tractable treatments of molecular properties. However, conventional hybrid functionals can be problematic for solids. Their nonlocal, Hartree-Fock-like exchange term decays slowly and incorporates unphysical features in metals and narrow-bandgap semiconductors. This article provides an overview of our group's work on designing hybrid functionals for solids. We focus on the Heyd-Scuseria Ernzerhof screened hybrid functional [J. Chem. Phys. 2003, 118, 8207], its applications to the chemistry and physics of solids and surfaces, and our efforts to build upon its successes. PMID- 19283262 TI - The inter-ring sigma/(pi/pi) covalent interactions of cyclodimes of benzenes. AB - To study the inter-ring sigma/(pi/pi) covalent interactions between non-radical pi-systems, five structures of cyclodimers of benzene (C6H6)2 with all the real frequencies, i.e. o-p'-dibenzene (), the pentacyclic dimer (), p-p'-dibenzene (), syn-o,o'-dibenzene (), and hexaprismane (), are obtained at the MP2/6-311G(d,p) level. Five inter-ring bonding mode types forming the inter-ring multicenter multielectron sigma/(pi/pi) covalent bonds are represented: , ring-edge type between a butterfly-shaped ring and a planar ring (4-center 4-electron bond); , edge-edge and ring-ring types between two identical butterfly-shaped rings (8 center 8-electron bond); , ring-ring type between two identical butterfly-shaped rings (4-center 4-electron bond); , edge-edge type between two identical planar rings (4-center 4-electron bond); and , face-face type between two identical planar rings (12-center 12-electron bond). The order of the large inter-ring interaction energies at the MP2/6-311+G(3d, 2p)+BF level is -99.15 (with two inter-ring C-C bonds)> -98.57 (with four C-C bonds)>-85.76 (with two C-C bonds)> 61.35 (with two C-C bonds)>-60.40 kcal mol(-1) (with six C-C bonds). However, this does not show an obvious relationship between the interaction energy and the number of the inter-ring C-C bonds. The reason is that the number of decisive influencing factors of the inter-ring interaction energy is not one but five: the number of the favorable inter-ring C-C single bonds, the number of the unfavorable four-membered rings themselves, the participating number of the four membered rings in unfavorable interaction among those rings, the number of the favorable non-planar melted six-membered ring, and the weak inter-ring pi/pi interaction (between two pi bonds in different rings). PMID- 19283263 TI - Quantum-mechanical wavepacket propagation in a sparse, adaptive basis of interpolating Gaussians with collocation. AB - We present an extension of our earlier work on adaptive quantum wavepacket dynamics [B. Hartke, Phys. Chem. Chem. Phys., 2006, 8, 3627]. In this dynamically pruned basis representation the wavepacket is only stored at places where it has non-negligible contributions. Here we enhance the former 1D proof-of-principle implementation to higher dimensions and optimize it by a new basis set, interpolating Gaussians with collocation. As a further improvement the Tnum approach from Lauvergnat and Nauts [J. Chem. Phys., 2002, 116, 8560] was implemented, which in combination with our adaptive representation offers the possibility of calculating the whole Hamiltonian on-the-fly. For a two dimensional artificial benchmark and a three-dimensional real-life test case, we show that a sparse matrix implementation of this approach saves memory compared to traditional basis representations and comes even close to the efficiency of the fast Fourier transform method. Thus we arrive at a quantum wavepacket dynamics implementation featuring several important black-box characteristics: it can treat arbitrary systems without code changes, it calculates the kinetic and potential part of the Hamiltonian on-the-fly, and it employs a basis that is automatically optimized for the ongoing wavepacket dynamics. PMID- 19283264 TI - Analyzing Kullback-Leibler information profiles: an indication of their chemical relevance. AB - The Kullback-Leibler information deficiency is evaluated along molecular internal rotational or vibrational coordinates and along the intrinsic reaction coordinate for several reactions (intra- and intermolecular proton transfer and SN2 reaction). For the first time an in depth analysis of the information deficiency along the reaction path is reported. The results are consistent with the Hammond postulate, indicating that the information profiles contain relevant chemical information. A local version of the information deficiency is defined by considering Hirshfeld's partitioning of atoms in molecules. The analysis of the local information profiles permits the identification of the atoms taking part in the electron reorganization processes. PMID- 19283265 TI - Infrared spectroscopic study of the effect of oleic acid on the deliquescence behaviour of ammonium sulfate aerosol particles. AB - In order to accurately assess the impact of fatty acids on the hygroscopic properties of atmospheric aerosol particles, (NH4)2SO4 (ammonium sulfate) and oleic acid (cis-9-octadecenoic acid) were chosen to perform this study as components of the particle phase. Micron-sized (700-900 nm) particles containing (NH4)2SO4 and oleic acid were generated by nebulising aqueous solutions of (NH4)2SO4 and sodium oleate. In this study, the effect of oleic acid on the deliquescence phase transition of particles was investigated in a room temperature aerosol flow tube (AFT) system using Fourier transform infrared (FTIR) spectroscopy. Particles morphologies and their chemical compositions were also analysed using a variety of techniques, including attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy and scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX). The deliquescence relative humidity (DRH) of the (NH4)2SO4 component, determined at 81+/-2%, was slightly lowered or not affected by the presence of different thickness of oleic acid (21 nm, 44 nm and 109 nm) present in the particles. Analyses of the results presented here are consistent with earlier studies about the possible effects of water-insoluble fatty acids coatings on the phase transitions of atmospheric aerosol particles. PMID- 19283266 TI - Time-resolved EPR investigation of [70]fulleropyrrolidine nitroxide isomers. AB - A novel [70]fulleropyrrolidine functionalized with a nitroxide radical has been synthesized. After pulsed photoexcitation, time-resolved electron paramagnetic resonance (EPR) spectra have been recorded in liquid solution at standard X-band (9.5 GHz) and W-band (95 GHz) microwave frequencies. The spectra exhibit strongly electron spin polarized ground and excited states, the latter being arising from the spin coupling of the nitroxide's electron spin with the fullerene's excited triplet state. The EPR parameters such as g-values and hyperfine coupling constants have been discussed in terms of the spin Hamiltonian for excited doublet, triplet and quartet states. On the basis of the strength of the exchange interaction between the fullerene triplet and the radical group, two, out of a possible four [70]fulleropyrrolidine isomeric monoadducts, have been characterized. PMID- 19283267 TI - Molecular dynamics simulations of the diffusion and rotation of Pt nanoclusters supported on graphite. AB - We study the diffusion and rotation of Pt nanoclusters supported on graphite. Using molecular dynamics simulation, we reveal that the supported Pt clusters have two types of bottom layers: a hexagonal lattice and a rectangular lattice. The diffusion rate of the Pt clusters mainly depends on the types of bottom layers. This observation is explained by the periodic lattice matching between the bottom layer and the graphite. In addition to lateral diffusion, we find that the Pt clusters with a rectangular or mixed lattice tend to rotate or wag on the z axis. By using the rotation rates and total movements of all atoms, we show that the rotation of a Pt cluster can be induced by the difference in the diffusion rates for the different types of bottom layers. This outcome is well in accord with our diffusion mechanism. PMID- 19283268 TI - Conformations, structural transitions and visible near-infrared absorption spectra of four-, five- and six-coordinated Cu(II) aqua complexes. AB - We have performed Car-Parrinello molecular dynamics simulations at ambient conditions for four-, five- and six-coordinated Cu(II) aqua complexes. The molecular geometry has been investigated in terms of Cu-O, Cu-H bond lengths and O-Cu-O bond angles and compared with earlier experimental measurement results and theoretical calculations. We find that the average Cu-O and Cu-H bond lengths increase with increasing coordination number. We have also observed relatively faster structural transition in the case of five-coordinated complex between trigonal bipyramidal and square pyramidal geometry. This result deviates from the findings of the earlier report (A. Pasquarello et al., Science, 2001, 291, 856) on copper(II) in aqueous solution and we attribute these differences to the neglect of solvent environment in our calculations. The averaged absorption spectra for the copper(II) aqua complexes have been computed using spin restricted density functional linear response formalism taking 100 snap shots from a trajectory of 0.48 ps. We find that the calculated spectra are significantly different, showing clear features that distinguish each coordination model. Comparison with the experimentally reported absorption spectra is made wherever it is possible and the results obtained favor the distorted fivefold-coordination arrangement for the molecular structure of the Cu(II) ion in aqueous solution. PMID- 19283269 TI - Volatile interface of biological oxidant and luminescent CdTe quantum dots: implications in nanodiagnostics. AB - In order to explore the feasibility of using quantum dots (QDs) as nanodiagnostic tools in cells under oxidative stress, the interaction of cysteine-functionalized CdTe QDs with peroxynitrite (PN), a powerful biological oxidant, was spectroscopically investigated. The photoluminescence (PL) of CdTe QDs was quenched to the extent of 85% at 86 microM concentration of peroxynitrite. In contrast, the loss of PL intensity was merely 7% for CdS QDs under identical conditions. The quenching pattern shows a positive deviation from Stern-Volmer equation, but obeys the Perrin equation for static quenching. The Perrin quenching constants increased with decreasing temperature. Size dependence was also prominently observed as quenching efficiency increased by 70% on moving from 4.0 nm sized CdTe QDs to 2.8 nm ones. Interestingly, the QDs exhibited a time dependent auto-recovery of photoluminescence. At 25 and 15 degrees C, the recovery time was found to be 1.25 and 3.5 h, respectively, however, the extent of recovery remained nearly the same, about 22%. Peroxynitrite followed a first order decay kinetics with a half-life of 45.6 min, which got substantially reduced to 2.9 min in the presence of CdTe QDs. The over-all mechanism of the interaction has been explained as a two-pronged attack of peroxynitrite on (i) the cysteine-capping layer and (ii) the CdTe core, resulting in a reversible and irreversible loss of QD-photoluminescence. PMID- 19283270 TI - Pressure-induced water flow through model nanopores. AB - This paper describes nonequilibrium molecular dynamics simulations of pressure induced transport of liquid water through model nanopores. We consider a simple model for a porous membrane consisting of a slab of water molecules held in a rigid ice structure and penetrated by a pore of nanometer scale dimensions. Both hydrophilic membranes composed of conventional TIP3P water and hydrophobic membranes consisting of modified water with the model partial charges set to zero are treated. Molecular dynamics simulation is employed to investigate the rate of water flow through the pore induced by a pressure difference across the membrane. The results are compared with the predictions of continuum hydrodynamics. We find that the flow rate of water through hydrophilic pores is much less than the continuum predictions, while the flux through hydrophobic pores can significantly exceed the continuum theory. Finally, we show asymmetric behavior in the flux vs. pressure difference for a conical nanopore, which thus acts as a Brownian ratchet. PMID- 19283271 TI - Blinking photoluminescence properties of single TiO2 nanodiscs: interfacial electron transfer dynamics. AB - Blinking photoluminescence was observed in single TiO2 nanodiscs (NDs) by using a laser scanning confocal microscope (LSCM)-coupled steady-state and ps-time resolved photoluminescence (PL) spectroscopic system, while it was not significantly observed for TiO2 quantum dots (QDs). Analysis of the PL blinking time trajectories revealed single-exponential kinetics with the average lifetimes of on-state (approximately 286 ms) and off-state (approximately 58 ms), implying the existence of inherent surface-trap sites which can be filled by photogenerated electron or hole. The PL spectra of single TiO2 NDs exhibited broad surface emissions with four decay times, which may be due to diffusion of the energies of electron or hole trap states related to surface structural changes by modification of TiO2 QDs. These results and the surface structural analysis (IR and XPS) suggests a simple model for the PL blinking of single TiO2 NDs that is based on repetitive interfacial electron transfer to the inherent surface trap sites (4Ti4+-OH) with Auger-assisted hole trapping in the multiple surface states as modified by the diffusive coordinate model and the surface-trap filling model. Based on this blinking mechanism and kinetics, the rates of the interfacial electron transfer and the back electron transfer in TiO2 NDs were determined to be 18 ns and 58 ms, respectively, which are slow enough to keep the polarization of e-h pairs at the surface for efficient photocatalysis and photovoltaic activities. The present methodology and results may be applicable to obtain surface exciton dynamics of various photoelectronic semiconductor nanostructures. PMID- 19283272 TI - The phase diagram of water at high pressures as obtained by computer simulations of the TIP4P/2005 model: the appearance of a plastic crystal phase. AB - In this work the high pressure region of the phase diagram of water has been studied by computer simulation by using the TIP4P/2005 model of water. Free energy calculations were performed for ices VII and VIII and for the fluid phase to determine the melting curve of these ices. In addition, molecular dynamics simulations were performed at high temperatures (440 K) observing the spontaneous freezing of the liquid into a solid phase at pressures of about 80,000 bar. The analysis of the structure obtained lead to the conclusion that a plastic crystal phase was formed. In the plastic crystal phase the oxygen atoms were arranged forming a body center cubic structure, as in ice VII, but the water molecules were able to rotate almost freely. Free energy calculations were performed for this new phase, and it was found that for TIP4P/2005 this plastic crystal phase is thermodynamically stable with respect to ices VII and VIII for temperatures higher than about 400 K, although the precise value depends on the pressure. By using Gibbs-Duhem simulations, all coexistence lines were determined, and the phase diagram of the TIP4P/2005 model was obtained, including ices VIII and VII and the new plastic crystal phase. The TIP4P/2005 model is able to describe qualitatively the phase diagram of water. It would be of interest to study if such a plastic crystal phase does indeed exist for real water. The nearly spherical shape of water makes possible the formation of a plastic crystal phase at high temperatures. The formation of a plastic crystal phase at high temperatures (with a bcc arrangements of oxygen atoms) is fast from a kinetic point of view occurring in about 2 ns. This is in contrast to the nucleation of ice Ih which requires simulations of the order of hundreds of ns. PMID- 19283273 TI - Triple points and coexistence properties of the dense phases of water calculated using computer simulation. AB - In recent years our group has performed a thorough study of the dense region of the phase diagram of water. In this paper we report the numerical results for the triple points and for the change in volume, Deltav, and enthalpy, DeltaH, along the coexistence lines involving liquid water and/or ices ih (hexagonal), II, III, V and VI for several simple water models. The predictions for Deltav using the TIP4P/2005 model are in excellent agreement with the experimental values. As to DeltaH for the same model, the computer simulation results are also satisfactory although there are small but significant differences between simulation and experiment. PMID- 19283274 TI - First-principles investigations on the functionalization of chiral and non-chiral carbon nanotubes by Diels-Alder cycloaddition reactions. AB - Functionalization of single-walled carbon nanotubes through cycloaddition reactions constitutes an effective route to obtain novel nanostructured materials with interesting properties. In this paper, we perform density functional theory calculations on Diels-Alder reactions at the sidewall of armchair, zigzag and chiral nanotubes by applying finite-length models of carbon nanotubes based on Clar's theory of the aromatic sextet. The analysis of binding energies and molecular orbitals suggests a prevalence of local factors, related to the structural and electronic properties at the coordinating site, in controlling the overall energetics of cycloaddition reactions. Our results can be expected to have strong implications in the development of rational strategies for the functionalization of carbon nanotubes of any stereochemistry. PMID- 19283275 TI - The structure of strontium-doped hydroxyapatite: an experimental and theoretical study. AB - First-principles modeling combined with experimental methods were used to study hydroxyapatite in which Sr2+ is substituted for Ca2+. Detailed analyses of cation oxygen bond distributions, cation-cation distances, and site 1-oxygen polyhedron twist angles were made in order to provide an atomic-scale interpretation of the observed structural modifications. Density functional theory periodic band structure calculations indicate that the Ca2+ to Sr2+ substitution induces strong local distortion on the hydroxyapatite lattice: the nearest neighbor Sr-O bond structures in both cationic sites are comparable to pure SrHA, while Sr induces more distortion at site 2 than site 1. Infrared vibrational spectroscopy (FTIR) and extended X-ray absorption fine structure (EXAFS) analysis suggest increasing lattice disorder and loss of OH with increasing Sr content. Rietveld refinement of synchrotron X-ray diffraction patterns shows a preference for the Ca1 site at Sr concentrations below 1 at.%. The ideal statistical occupancy ratio Sr2/Sr1=1.5 is achieved for approximately 5 at.%; for higher Sr concentrations occupation of the Ca2 site is progressively preferred. PMID- 19283276 TI - Effect of the acidic strength on the vapor phase Beckmann rearrangement of cyclohexanone oxime over the MFI zeolite: an embedded ONIOM study. AB - The mechanism and energetic profile of the Beckmann rearrangement reaction of cyclohexanone oxime to epsilon-caprolactam catalyzed by the H-[Al]-MFI and H-[B] MFI zeolites were investigated by both the bare cluster and the ONIOM models at the B3LYP/6-31G(d,p) and the B3LYP/6-31G(d,p):MNDO levels of theory, respectively. In order to improve the energetic properties and take into account the whole zeolite framework effect, single point calculations are undertaken at the embedded ONIOM2 schemes; MP2/6-311G(d,p):HF/6-31G(d) with an additional long range electrostatic potential from the extended zeolite framework. The reaction mechanism of the Beckmann rearrangement over the acid site of zeolites consists of three steps: the 1,2 H shift, the rearrangement and the tautomerization. The activation energies for the Beckmann rearrangement of cyclohexanone oxime on the H-[Al]-MFI zeolite are calculated to be 31.46, 16.15 and 18.95 kcal mol(-1), for the first, second and third steps, respectively, whereas in the H-[B]-MFI zeolite, the energy barriers for each step of the reaction are 24.33, 7.46 and 20.43 kcal mol(-1), respectively. The rate-determining step of the reaction is the first step, which is the transformation from the N-ended cyclohexanone oxime adsorption complex and the O-ended one. These results signify the important role that the acid strength of zeolites plays in altering the energy profile of the reaction. The results further indicate that the weak Bronsted acid sites in the [B]-MFI zeolite could better catalyze the Beckmann rearrangement of cyclohexanone oxime than the strong acid sites in the [Al]-MFI zeolite, as compared with the quantitatively low activation energy of most steps. However, the turnover reaction of the H-[B]-MFI zeolite might be delayed by the quantitatively high desorption energy of the product as compared to the adsorption energy of the reactant. PMID- 19283277 TI - Periodic local-MP2 computational study of crystalline neon. AB - Face-centered-cubic crystalline Neon is taken as a test system to explore the influence of computational parameters on the quality of the MP2 solution provided by the Cryscor program using a local-correlation approach. The effect of the various approximations adopted is analyzed: basis set limitations, finite size of excitation domains, truncation of the tails of the local functions, approximate evaluation of two-electron integrals, estimate (by extrapolation) of long-range contributions are shown to play roles of different importance. The Ne2 dimer is used as an auxiliary test case in order to allow comparison with recent and accurate literature data. PMID- 19283278 TI - Modifications induced by pretreatments on Au/SBA-15 and their influence on the catalytic activity for low temperature CO oxidation. AB - SBA-15 functionalization with mercaptopropyltrimethoxysilane has been used to prepare supported gold catalysts for the low temperature CO oxidation reaction. Supports and catalysts have been characterized by chemical analysis, CHS analysis, XRD, TGA, nitrogen adsorption-desorption at 77 K, TEM, CPMAS NMR, XPS and EPR. Catalytic runs have been carried out at atmospheric pressure and 313-623 K and the influence of diverse thermal treatments of the samples prior to reaction has been investigated. The presence of organic residues and the size of the gold nanoparticles strongly affect catalytic activity. Only high-temperature calcination in air followed by treatment under H2 atmosphere leads to active catalysts. After complete elimination of the functionalizing agent, caused by the calcination step, a gold-mediated "activation" process of the silica support takes place during the hydrogen treatment. As a consequence, active catalysts for the low temperature CO oxidation are obtained, even though the size of the Au particles is too large for establishing direct Au-oxygen interactions, usually assumed to be essential for the reaction over silica-supported gold catalysts. PMID- 19283279 TI - Single amino acid chelates (SAAC): a strategy for the design of technetium and rhenium radiopharmaceuticals. AB - Radiolabeled biomolecules can be used to visualize a variety of diseases through interaction with specific cell receptors. A key step is the introduction of a molecular entity that allows facile labeling with the medically useful radionuclide (99m)Tc without significant alteration of the structure and function of the biomolecule. One strategy focuses on the design of single amino acid chelates (SAACs), novel bifunctional chelators constructed from derivatized amino acids or amino acid analogues. The chelating terminus of the SAAC has been designed for effective coordination to the {(99m)Tc(CO)(3)}(+) core, while the other terminus allows incorporation into any position along a peptide sequence or into a variety of biomolecules. In applications to peptidic materials, the approach affords significant flexibility in the choice of donors for (99m)Tc coordination combined with the considerable advantages of routine solid phase synthetic techniques. The methodology allows libraries of peptide-based (99m)Tc(i) and (186,188)Re(i) radiopharmaceuticals to prepared using conventional automated peptides synthesis. Other biomolecules, including nucleosides, carbohydrates, folic acid and vitamin B12 are also readily modified using analogous methods. The approach also allows the preparation of isostructural (99m)Tc and Re complexes for the correlation of in vivo and in vitro imaging studies. PMID- 19283280 TI - Anion receptors that contain metals as structural units. AB - Transition metals can be used as structural elements to build up anion receptors, enhancing H-bond donor tendencies, favouring the assembling of the molecular framework, inducing the formation of a cage. The versatile spectroscopic and electrochemical properties of the metal ion can provide a convenient signalling mechanism to communicate the occurrence of anion recognition. PMID- 19283281 TI - Dye displacement assay for saccharide detection with boronate hydrogels. AB - Hydrogel spheres, fashioned from an operationally simple mould, that incorporate boronate units were shown to function as saccharide sensors. PMID- 19283282 TI - Reversible increase in the redox potential of cytochrome c in methanol. AB - The E degrees ' of cytochrome c on a self-assembled monolayer (SAM) modified gold electrode increased by 300 mV immediately on immersion in methanol; on re immersion of the electrode in aqueous buffer, the original faradaic response was restored, but over a period of 120 min, indicating that methanol causes a significant change in conformation/orientation of the protein. PMID- 19283283 TI - Towards quorum-quenching catalytic antibodies. AB - The development of a novel method to attenuate bacterial virulence is reported, which is based upon the use of designed transition-state analogues to select human catalytic antibodies capable of degrading bacterial quorum-sensing molecules. PMID- 19283284 TI - Mesostructured material based on [Re6Te8(CN)6]4- cluster and Mn(2+): a rational synthesis of hexagonal nonoxidic mesoscale material. AB - A new nonoxidic mesostructured material with highly ordered hexagonal symmetry, [C(16)H(33)N(CH(3))(3)](3)Mn(0.5)[Re(6)Te(8)(CN)(6)], has been obtained through supramolecular assembly of [Re(6)Te(8)(CN)(6)](4-) clusters in the presence of a transition metal cation, Mn(2+), with cetyltrimethylammonium bromide as a liquid crystal template under hydrothermal conditions. PMID- 19283285 TI - A [Mn18Dy] SMM resulting from the targeted replacement of the central MnII in the S = 83/2 [Mn19]-aggregate with DyIII. AB - Anisotropy can be introduced into the [Mn(19)]-aggregate, which currently has the highest known spin ground state of S = 83/2, by the targeted replacement of the central Mn(II) cation with Dy(III) leading to a [Mn(18)Dy] complex with the same core topology showing slow relaxation of the magnetisation. PMID- 19283286 TI - Relaying stereochemistry through aromatic ureas: 1,9 and 1,15 remote stereocontrol. AB - The well-defined conformation of an N,N'-diarylurea allows a chiral sulfinyl substituent to influence diastereoselectivity in the formation of new stereogenic centres up to 14 bond lengths away. PMID- 19283287 TI - Catalytic electrochemistry of a [NiFeSe]-hydrogenase on TiO2 and demonstration of its suitability for visible-light driven H2 production. AB - A [NiFeSe]-hydrogenase able to produce H(2) in the presence of O(2) forms the basis of a hybrid enzyme-TiO(2) nanoparticle system with a co-attached synthetic Ru photosensitiser for visible-light driven H(2) production at room temperature from neutral water under non-strictly anaerobic conditions on the bench. PMID- 19283288 TI - A 'sting' on Grubbs' catalyst: an insight into hydride migration between boron and a transition metal. AB - An unusual ruthenium(ii) complex frozen at an intermediate point of hydride transfer between boron and ruthenium centres is reported. PMID- 19283289 TI - High-throughput trace melamine analysis in complex mixtures. AB - Ambient ionization using a low-temperature plasma (LTP) probe combined with tandem mass spectrometry (MS/MS) allows detection and quantitation of melamine in milk powder, whole milk and other products at levels down to low ppb in analysis times of a few tens of seconds. PMID- 19283290 TI - Rapid detection of melamine in untreated milk and wheat gluten by ultrasound assisted extractive electrospray ionization mass spectrometry (EESI-MS). AB - A novel method based on ultrasound-assisted EESI-MS has been developed and applied to rapidly detect the presence of melamine in raw milk, wheat gluten and milk powder with no or minor sample pre-treatment; the high sample throughput and figures of merit make it specially useful for screening melamine levels well below the current safety limit in various food matrices. PMID- 19283291 TI - The crystal structure of beta-RDX-an elusive form of an explosive revealed. AB - The crystal structure of the highly metastable beta-form of RDX shows that the molecules adopt different conformations compared to the alpha-form and that, contrary to previous reports, the beta-form obtained at ambient pressure is not the same form as that obtained at elevated temperatures and pressures. PMID- 19283292 TI - Simplified methods for the functionalisation of 3-hexoxythiophenes at the 5 position and further reactions to alkynyl and vinyl derivatives. AB - Kinetically controlled lithiation of 3-hexoxythiophene in the 5-position allows the synthesis of interesting functionalised building blocks after reaction with N,N-dimethylformamide and iodine. PMID- 19283293 TI - Light-activation of gene function in mammalian cells via ribozymes. AB - A ribozyme based gene control element enabled the spatio-temporal regulation of gene function in mammalian cell culture with light. PMID- 19283294 TI - Dynamic kinetic resolution of beta-keto sulfones via asymmetric transfer hydrogenation. AB - The dynamic kinetic resolution of beta-keto sulfones was achieved via asymmetric transfer hydrogenation using (S,S)-RuCl[N-(tosyl)-1,2-diphenylethylenediamine](p cymene) in the presence of formic acid and triethylamine afforded the desired products in good yield with up to >99 : 1 dr and high ee up to >99%. PMID- 19283295 TI - Thermal cyclotrimerization of tetraphenyl[5]cumulene (tetraphenylhexapentaene) to a tricyclodecadiene derivative. AB - Tetraphenyl[5]cumulene (tetraphenylhexapentaene) underwent cyclotrimerization in refluxing toluene for 10-15 min to give a tricyclodecadiene derivative in 68% yield. PMID- 19283296 TI - Asymmetric heterometal string complexes: stereochemical control of the unique isomer of (4,0)[CuCuPd(npa)4Cl][PF6] and (4,0)[CuCuPt(npa)4Cl][PF6]. AB - This paper describes the synthesis and physical properties of a unique metal string complex isomer containing an asymmetric heterometallic backbone. PMID- 19283297 TI - G-Quadruplex-based DNAzyme as a sensing platform for ultrasensitive colorimetric potassium detection. AB - Two G-quadruplex DNAs are able to form peroxidase-like DNAzymes selectively promoted by K(+), which provides a label-free approach to colorimetric detection of K(+) with the highest sensitivity ever reported. PMID- 19283298 TI - Outer-sphere amidopyridyl extractants for zinc(ii) and cobalt(ii) chlorometallates. AB - Protonated amidopyridyl ligands show high selectivity for the extraction of [CoCl(4)](2-) or [ZnCl(4)](2-) over chloride ion into organic media via the formation of N-H and C-H hydrogen-bond donors to the outer coordination spheres of the chlorometallates. PMID- 19283299 TI - Casting heteracalixarenes from calixarene templates: a unique synthetic strategy. AB - A simple, intuitive and modular strategy to synthesize higher heteracalixarene (in particular thiacalixarene) homologs using respective calixarene templates has been developed and demonstrated. PMID- 19283300 TI - Regioselective access to 3I-O-substituted-beta-cyclodextrin derivatives. AB - Formation of a copper(ii)-beta-cyclodextrin (beta-CD) complex in an aqueous medium allowed the regioselective introduction into the oligosaccharide of a benzyl or a bromo-allyl substituent at O-3. PMID- 19283301 TI - A thermal 6pi electrocyclization strategy towards taiwaniaquinoids. First enantiospecific synthesis of (-)-taiwaniaquinone G. AB - The first route towards taiwaniaquinoid terpenes bearing an A/B trans configuration has been developed through a sequence which includes a thermal 6pi electrocyclization. PMID- 19283302 TI - Synthesis and isotopic labeling of a naturally occurring alkyl-thiadiamondoid. AB - Bridgehead-methylated 2-thioadamantane has been synthesized in a new efficient way. PMID- 19283303 TI - Synergic asymmetric organocatalysis (SAOc) of Cinchona alkaloids and secondary amines in the synthesis of bicyclo[2.2.2]octan-2-ones. AB - A synergistic effect between chiral secondary and tertiary amines gives access to bicyclo[2.2.2]octan-2-ones with up to 90% ee and >10 : 1 dr. PMID- 19283304 TI - Iron-catalysed Negishi coupling of benzyl halides and phosphates. AB - Iron-based catalysts containing either 1,2-bis(diphenylphosphino)benzene or 1,3 bis(diphenylphosphino)propane give excellent activity and good selectivity in the Negishi coupling of aryl zinc reagents with a range of benzyl halides and phosphates. PMID- 19283306 TI - A note on estimating treatment effect for time-to-event data in a literature based meta-analysis. AB - OBJECTIVES: In a literature-based meta-analysis for time-to-event data, the hazard ratio in each trial is often estimated from the summary statistics described in the article. Several methods have been proposed: the direct method (Peto method); the indirect method using a p-value by the log-rank test and the number of total events; and the survival curve method using the Kaplan-Meier estimate. However, there has been no published report on a detailed investigation of these methods. We evaluated the performance of these methods by simulation. METHODS: In a set of simulation experiments, performance of five methods was evaluated by the bias of estimated log hazard ratio and coverage probability of the confidence interval. The methods evaluated were: 1) Cox regression analysis, 2) direct method, 3) indirect method, 4) survival curve method, and 5) modified survival curve method with an alternative weighting scheme. RESULTS: The direct method was confirmed to have a high degree of accuracy. Although the indirect method was also highly accurate, it tended to underestimate effect size when there was a strong effect. The survival curve method tended to underestimate effect size when event numbers were small and effect size was large. The modified survival curve method could alleviate this tendency toward underestimation of effect size found with the original survival curve method. CONCLUSIONS: When the Kaplan-Meier curve is used to estimate hazard ratios in trials with small sample size in the literature-based meta-analysis, we should check critically whether those trials' hazard ratios and overall hazard ratio are underestimated or not. PMID- 19283307 TI - Computer simulation of coronary flow waveforms during caval occlusion. AB - OBJECTIVES: Mathematical modeling of the cardiovascular system is a powerful tool to extract physiologically relevant information from multi-parametric experiments. The purpose of the present work was to reproduce by means of a computer simulator, systemic and coronary measurements obtained by in vivo experiments in the pig. METHODS: We monitored in anesthetized open-chest pig the phasic blood flow of the left descending coronary artery, aortic pressure, left ventricular pressure and volume. Data were acquired before, during, and after caval occlusion. Inside the software simulator (CARDIOSIM) of the cardiovascular system, coronary circulation was modeled in three parallel branching sections. Both systemic and pulmonary circulations were simulated using a lumped parameter mathematical model. Variable elastance model reproduced Starling's law of the heart. RESULTS: Different left ventricular pressure-volume loops during experimental caval occlusion and simulated cardiac loops are presented. The sequence of coronary flow-aortic pressure loops obtained in vivo during caval occlusion together with the simulated loops reproduced by the software simulator are reported. Finally experimental and simulated instantaneous coronary blood flow waveforms are shown. CONCLUSIONS: The lumped parameter model of the coronary circulation, together with the cardiovascular system model, is capable of reproducing the changes during caval occlusion, with the profound shape deformation of the flow signal observed during the in vivo experiment. In perspectives, the results of the present model could offer new tool for studying the role of the different determinants of myocardial perfusion, by using the coronary loop shape as a "sensor" of ventricular mechanics in various physiological and pathophysiological conditions. PMID- 19283308 TI - periodR - an R package to calculate long-term cancer survival estimates using period analysis. AB - OBJECTIVE: In this paper, a software package for the R language and system for statistical computing is presented for computation of long-term cancer survival estimates based on the period analysis approach. The period analysis approach provides up-to-date long-term survival estimates of concurrently diagnosed patients, enables early detection of recent changes in long-term prognosis of cancer patients and provides better survival predictions for recently and currently diagnosed patients than traditional cohort-based approaches. METHODS: Computation of absolute and relative survival estimates (both conditional follow up year-specific and cumulative survival estimates) and their standard errors is based on standard actuarial methodology. For relative survival estimation the "Ederer II" and "Hakulinen" method were implemented. RESULTS: The package may be used for period analysis as well as traditional cohort-based survival estimation. The package further provides functions for the export of survival estimates for use with spreadsheet programs and for plotting survival curves. The application of period analysis is illustrated using stomach cancer data included in the package. CONCLUSION: Application of period analysis has gradually increased in recent years but continues to be limited by availability of affordable and easy to-use software tools. The presented R package aims at closing this gap and will further facilitate the use of period analysis for the research community working with population-based cancer registry data. The software is freely available for download on the website of the Saarland Cancer Registry at http://www.krebsregister.saarland.de/improve/periodR_en.html. PMID- 19283309 TI - Comparative evaluation of balancing properties of stratified randomization procedures. AB - OBJECTIVES: If in a clinical trial prognostic factors are known in advance to be associated with the outcome of a patient it is often recommended that the randomization for a clinical trial should be stratified on these factors, particularly in a multicenter trial. Unfortunately, stratified or covariate adaptive randomization does not always promote greater balance between the numbers of treatment assignments to A and B within each stratum and thus overall. Because such designs have numerous parameters that must be specified, simulation is a good tool to investigate the impact of these parameters on balance. METHODS: We investigate and discuss in more detail the difference in balancing performance of three stratified randomization procedures. The permuted-block randomization within strata, the "minimization" method and "self-adjusting" design are assessed overall, within levels of prognostic factors, and within strata. RESULTS: We show the superior performance of "self-adjusting" design and the extent of balancing losses occurring with permuted-block randomization within levels of factors and with "minimization" within strata. A summary of principal conclusions regarding the balancing properties of stratified randomization procedures is presented and general recommendations are offered. CONCLUSIONS: The relative merits of each procedure should be weighted carefully in relation to the characteristics of the trial. Considering the likelihood of imbalances, the sample size and values of parameters of stratified randomization procedures (number of prognostic factors, number of factor levels, block size) are important when choosing a randomization procedure. PMID- 19283310 TI - Evaluating the impact of hybrid electronic-paper environments upon novice nurse information seeking. AB - OBJECTIVE: The purpose of this study was to investigate the effects of hybrid environments (i.e. where part of the patient record is paper-based and part of it is electronic) upon aspects of novice nurse information seeking (i.e. amount of information accessed, choice of key information sources, type of information and use of information seeking tactics). METHODS: A within group, laboratory, experimental study was conducted using two simulated environments (i.e. a paper environment and a hybrid environment). Thirty-five novice nurses participated in the study. RESULTS: Findings revealed significant differences between the paper and hybrid environments in terms of their effects upon aspects of novice nurse information seeking. Subjects accessed: 1) less information in the hybrid environment than the paper environment, 2) more non-electronic sources of information were accessed by novice nurses in the hybrid environment, and 3) novice nurses used more passive information seeking tactics in the hybrid environment than the paper environment. Qualitative findings from the cued recall data revealed subjects experienced increased cognitive load in the hybrid environment. CONCLUSIONS: Hybrid environments may affect aspects of novice nurse information seeking. Future research should explore the effects of hybrid environments upon the information seeking of other types of health professionals (e.g. physicians, physiotherapists) with differing levels of expertise (i.e. novice, intermediate and expert). PMID- 19283311 TI - Consent-based access to core EHR information. Collaborative approaches in Norway. AB - OBJECTIVE: Lack of access to updated drug information is a challenge for healthcare providers in Norway. Drug charts are updated in separate EHR systems but exchange of drug information between them is lacking. In order to provide ready access to updated medication information, a project for consent-based access to a core EHR has been established. METHODS: End users have developed requirements for additions to the medication modules in the EHR systems in cooperation with vendors, researchers and standardization workers. The modules are then implemented by the vendors, tested in the usability lab, and finally tested by the national testing and approval service before implementation. An ethnographic study, with focus on future users and their interaction with other actors regarding medicines and medication, has included semi-/unstructured interviews with the involved organizational units. RESULTS: The core EHR uses the EHR kept by the patient's regular GP as the main source of information. A server based solution has been chosen in order to keep the core EHR accessible outside the GP's regular work hours. The core EHR is being tested, and the EHR-vendors are implementing additions to their systems in order to facilitate communication with the core EHR. All major EHR-system vendors in Norway participate in the project. CONCLUSIONS: The core EHR provides a generic basis that may be used as a pilot for a national patient summary. Examples of a wider use of the core EHR can be: shared individual plans to support continuity of care, summary of the patient's contacts with health providers in different organizations, and core EHR information such as important diagnoses, allergies and contact information. Extensive electronic cooperation and communication requires that all partners adjust their documentation practices to fit with other actors' needs. The implementation effects on future work practices will be followed by researchers. PMID- 19283312 TI - Automatic acquisition of synonym resources and assessment of their impact on the enhanced search in EHRs. AB - OBJECTIVE: Currently, the use of natural language processing (NLP) approaches in order to improve search and exploration of electronic health records (EHRs) within healthcare information systems is not a common practice. One reason for this is the lack of suitable lexical resources. Indeed, in order to support such tasks, various types of such resources need to be collected or acquired (i.e., morphological, orthographic, synonymous). METHODS: We propose a novel method for the acquisition of synonymy resources. This method is language-independent and relies on existence of structured terminologies. It enables to decipher hidden synonymy relations between simple words and terms on the basis of their syntactic analysis and exploitation of their compositionality. RESULTS: Applied to series of synonym terms from the French subset of the UMLS , the method shows 99% precision. The overlap between thus inferred terms and the existing sparse resources of synonyms is very low. In order to better integrate these resources in an EHR search system, we analyzed a sample of clinical queries submitted by healthcare professionals. CONCLUSIONS: Observation of clinical queries shows that they make a very little use of the query expansion function, and, whenever they do, synonymy relations are rarely involved. PMID- 19283313 TI - Development of a national core dataset for preoperative assessment. AB - OBJECTIVE: To define a core dataset for preoperative assessment to leverage uniform data collection in this domain. This uniformity is a prerequisite for data exchange between care providers and semantic interoperability between health record systems. METHODS: To design this core dataset a combination of literature review and expert consensus meetings were used. In the first meeting a working definition for "core dataset" was specified. Subgroups were formed to address major headings of the core dataset. In the following eight meetings data items for each subheading were discussed. The items in the resulting draft of the dataset were compared to those retrieved from an earlier literature review study. In the last two expert meetings modifications of the dataset were performed based on the result of this literature study. RESULTS: Based on expert consensus a draft dataset including 82 data items was designed. Seventy-six percent of data items in the draft dataset were covered by the literature study. Nine data items were modified in the draft and 14 data items were added to the dataset based on input from the literature review. The final dataset of 93 data items covers patient history, physical examination, supplementary examination and consultation, and final judgment. CONCLUSIONS: This preoperative-assessment dataset was defined based on expert consensus and literature review. Both methods proved to be valuable and complementary. This dataset opens the door for creating standardized approaches in data collection in the preoperative assessment field which will facilitate interoperability between different electronic health records and different users. PMID- 19283314 TI - Analysis and evaluation of EHR approaches. AB - OBJECTIVES: EHR systems are core applications in any eHealth/pHealth environment and represent basic services for health telematics platforms. Standards Developing Organizations as well as national programs define EHR architectures as well as related design, implementation, and deployment processes. Claiming to meet the challenge for semantic interoperability and to offer a sustainable pathway, the resulting documents and specifications are sometimes controversial and even inconsistent. METHODS: Based on long-term experiences from national and international EHR projects, inputs from related academic groups, and active involvement at CEN, ISO, HL7, an analysis and evaluation study has been performed. Using the Generic Component Model (GCM) reference architecture, the characteristics for advanced and sustainable EHR architectures have been investigated. The dimensions of such an architectural reference model have been described, including basic principles of the underlying formal logical framework. RESULTS: Strengths and weaknesses of the different standards, specifications, and approaches have been studied and summarized. Migration pathways for re-using and harmonizing the available materials as well as for formally defining standards development roadmaps can be derived. CONCLUSIONS: For providing interoperable and sustainable EHR systems, an EHR architecture reflecting all paradigms of the GCM is absolutely necessary. The resulting EHR solution represents a services architecture of distributed components. The development process shall be completely model-driven and tool-based with formalized specifications of all domains' aspects. PMID- 19283315 TI - Enhanced semantic interoperability by profiling health informatics standards. AB - OBJECTIVES: Several standards applied to the healthcare domain support semantic interoperability. These standards are far from being completely adopted in health information system development, however. The objective of this paper is to provide a method and suggest the necessary tooling for reusing standard health information models, by that way supporting the development of semantically interoperable systems and components. METHODS: The approach is based on the definition of UML Profiles. UML profiling is a formal modeling mechanism to specialize reference meta-models in such a way that it is possible to adapt those meta-models to specific platforms or domains. A health information model can be considered as such a meta-model. RESULTS: The first step of the introduced method identifies the standard health information models and tasks in the software development process in which healthcare information models can be reused. Then, the selected information model is formalized as a UML Profile. That Profile is finally applied to system models, annotating them with the semantics of the information model. The approach is supported on Eclipse-based UML modeling tools. The method is integrated into a comprehensive framework for health information systems development, and the feasibility of the approach is demonstrated in the analysis, design, and implementation of a public health surveillance system, reusing HL7 RIM and DIMs specifications. CONCLUSIONS: The paper describes a method and the necessary tooling for reusing standard healthcare information models. UML offers several advantages such as tooling support, graphical notation, exchangeability, extensibility, semi-automatic code generation, etc. The approach presented is also applicable for harmonizing different standard specifications. PMID- 19283316 TI - Definitions and qualifiers in SNOMED CT. AB - OBJECTIVES: An important feature of SNOMED CT is post-coordination, which is enabled by the SNOMED CT representation specifying whether a relationship is a defining or a qualifier relationship. In this paper the use of qualifier relationships in SNOMED CT is analyzed, as well as the extent to which qualifiers interact with defining relationships, so that pre-coordinated concepts can also be post-coordinated. METHODS: The July 2007 release of SNOMED CT was imported into a database. Analyses were performed by querying this database. RESULTS: Qualifier relationships occur in 10 out of 61 types of attribute relationships, and it is shown that generally pre-coordinated concepts cannot be constructed by applying post-coordination using qualifier relationships. Most of the qualifier relationships have generic target concepts, making it possible to construct concepts which are not clinically sensible. A logic-based representation is proposed to overcome the drawbacks of the current model. CONCLUSIONS: Defining and qualifier relationships both enable post-coordination in SNOMED CT. Introducing qualifiers for more types of relationships, and using qualifier relationships with more specific target concepts will further improve post coordination in SNOMED CT. PMID- 19283317 TI - Granularity issues in the alignment of upper ontologies. AB - OBJECTIVES: The application of upper ontologies has been repeatedly advocated for to support the interoperability between different domain ontologies for facilitating the shared use of data within and across disciplines. BioTop is an upper domain ontology that aims at aligning more specialized biomolecular and biomedical ontologies. The integration of BioTop and the upper ontology Basic Formal Ontology (BFO) is the objective of this study. METHODS: BFO was manually integrated into BioTop, observing both its free text and formal definitions. BioTop classes were attached to BFO classes as children and BFO classes were reused in the formal definitions of BioTop classes. A description logics reasoner was used to check the logical consistency of this integration. The domain adequacy was checked manually by domain experts. RESULTS: Logical inconsistencies were found by the reasoner when applying the BFO classes for fiat and aggregated objects in some of the BioTop class definitions. We discovered that the definition of those particular classes in BFO was dependent on the notion of physical connectedness. Hence we suggest ignoring a BFO subbranch in order not to hinder cross-granularity integration. CONCLUSION: Without introducing a more sophisticated theory of granularity, the described problems cannot be properly dealt with. Whereas we argue that an upper ontology should be granularity independent, we illustrate how granularity-dependent domain ontologies can still be embedded into the framework of BioTop in combination with BFO. PMID- 19283318 TI - The impact of SOA for achieving healthcare interoperability. An empirical investigation based on a hypothetical adoption. AB - OBJECTIVES: The evaluation of a service-oriented prototype implementation for healthcare interoperability. METHODS: A prototype framework was developed, aiming to exploit the use of service-oriented architecture (SOA) concepts for achieving healthcare interoperability and to move towards a virtual patient record (VPR) paradigm. The prototype implementation was evaluated for its hypothetical adoption. The evaluation strategy was based on the initial proposition of the DeLone and McLean model of information systems (IS) success [1], as modeled by Iivari [2]. A set of SOA and VPR characteristics were empirically encapsulated within the dimensions of IS success model, combined with measures from previous research works. The data gathered was analyzed using partial least squares (PLS). RESULTS: The results highlighted that system quality is a partial predictor of system use but not of user satisfaction. On the contrary, information quality proved to be a significant predictor of user satisfaction and partially a strong significant predictor of system use. Moreover, system use did not prove to be a significant predictor of individual impact whereas the bi-directional relation between use and user satisfaction did not confirm. Additionally, user satisfaction was found to be a strong significant predictor of individual impact. Finally, individual impact proved to be a strong significant predictor of organizational impact. CONCLUSIONS: The empirical study attempted to obtain hypothetical, but still useful beliefs and perceptions regarding the SOA prototype implementation. The deduced observations can form the basis for further investigation regarding the adaptability of SOA implementations with VPR characteristics in the healthcare domain. PMID- 19283319 TI - The importance of medical and nursing sub-cultures in the implementation of clinical information systems. AB - OBJECTIVE: To measure doctors' and nurses' perceptions of organisational culture and relate this to their attitudes to, and satisfaction with, a hospital-wide mandatory computerised provider order entry (CPOE) system in order to illuminate cultural compositions in CPOE use. METHODS: A cross-sectional survey design was employed. Data were collected by administering an organisational culture survey (Organisational Culture Inventory, OCI) along with a user-satisfaction survey to a population of 103 doctors and nurses from two clinical units in an Australian metropolitan teaching hospital. Clinicians from the hospital had used the CPOE system since 1991 to order and view clinical laboratory and radiology tests electronically for all patients. The OCI provides a measure of culture in terms of three general styles which distinguish between: constructive; passive/defensive, and aggressive/defensive cultures. The cluster which best describes the overall culture is the one that has the highest percentile score when the percentile scores of the four cultural norms included in the cluster are averaged. The user satisfaction survey asked questions relating to satisfaction with, and attitudes to, the system. RESULTS: We found identifiable sub-cultures based on professional divisions where doctors perceived an aggressive-defensive culture (mean percentile score = 43.8) while nurses perceived a constructive culture (mean percentile score = 61.5). There were significant differences between doctors and nurses on three of the attitude variables with nurses expressing more positive views towards CPOE than doctors. CONCLUSION: The manifestation of sub-cultures within hospitals and the impact this has on attitudes towards clinical information systems should be recognized and addressed when planning for system implementation. Identification and management of the cultural characteristics of different groups of health professionals may facilitate the successful implementation and use of clinical information systems. PMID- 19283320 TI - Categorizing communication errors in integrated hospital information systems. AB - OBJECTIVES: Hospital information systems (HIS) are the hospital's nervous system. The HIS are mostly grown over several years, dedicated to specific needs and comprise individual workarounds. Changes to such complex systems may cause a variety of differ-ent negative side-effects. In order to understand the nature of incorrect communication in integrated HIS, a concise structured categorization of common communication problems and their reasons is essential. The objective of this paper is to present such a categorization, its development and verification. METHODS: We used a combined approach for the development of the error categorization: We started with a qualitative content analysis on available literature in PubMed. In order to ensure the validity and completeness of the results, we chose the method of problem-centered expert interviews. RESULTS: The resulting categorization of communication problems is represented as a five-level hierarchy. It comprises 81 problems that are related to the electronic communication. Further, it contains in total 229 entries that are either the reasons of these problems or recommendation for avoiding the problems. CONCLUSION: To our knowledge there is no similar summary that concisely summarizes common communication problems and also refers to their underlying reasons. Equivalent content is mostly published in experience reports that just concentrate on single aspects. We used the details of such references in order to compile our categorization - it thus can be regarded as an intersection of relevant experiences. The categorization can raise a basic awareness on potential problems and supports the understanding of the underlying reasons. An evaluation in a real environment must prove whether the content of the categorization is correct. PMID- 19283321 TI - Test turnaround times and mortality rates 12 and 24 months after the introduction of a computerised provider order entry system. AB - OBJECTIVES: Few studies have measured the long-term effects of computerised provider order entry systems on pathology test turnaround time. Further, a recent study has raised the possibility that such systems, which require significant work practice change, may be associated with an increase in mortality rates. Our study answered two questions in relation to system introduction in a major Australian teaching hospital: i) are improvements in turnaround times achieved in the first 12 months after system introduction sustained 24 months post implementation; and ii) do mortality rates change following the introduction of an order entry system? METHODS: Turnaround time and mortality rates 5 months before and 12 and 24 months after implementation of a computerised order entry system were measured. Turnaround time was defined as the time from receipt of a specimen and order in a laboratory to availability of a result. RESULTS: Improvements in turnaround time achieved in the first 12 months were sustained with a further significant 12.6% reduction at 24 months post-implementation, with no change in average number of tests per patient. The mortality rate significantly increased in the year following system introduction but returned to the pre-system rate in the second year of system use. Review of the excess deaths demonstrated these were most likely attributable to a coincidental influenza outbreak and not to system introduction. CONCLUSIONS: The computerised provider order entry system produced sustained and continuing improvements in laboratory efficiency over a two-year period. Associations between increased mortality rates and system introduction should be investigated carefully to ascertain any likely association. PMID- 19283322 TI - Reproducible extraction of local and global parameters for functional analysis of the left ventricle in 4D MR image data. AB - OBJECTIVES: Left ventricle (LV) segmentation is required to quantify LV volume and mass parameters. Therefore, spatiotemporal Cine MR sequences in the short and long axis of the heart are acquired. Generally, LV segmentation methods consider short-axis sequences only. The reduced resolution in long-axis direction is one of the main reasons for inaccurate parameter extraction in the apical and basal area. The segmentation approach presented combines short- and long-axis information as well as motion tracking to enable the functional LV analysis in 4D MR Image Data. METHODS: First, anatomical landmarks like the mitral valve and the apex are defined in long-axis views in diastolic and systolic phase in order to specify the upper and lower boundary of the LV. Second, motion field approximation using non-linear registration enables the automatic contour propagation to all time points. Third, intersection planes are defined parallel to the mitral valve plane covering the whole ventricle. Finally, the 4D LV surface model is generated appending all in-plane contours. The segmentation results in short-axis images are checked and adjusted interactively and quantitative parameters are extracted. RESULTS: For evaluation the contours of 19 different datasets were traced by two medical experts using a contour drawing tool and the new segmentation tool. The results were compared to evaluate automatic contour propagation, robustness of the segmentation as well as interaction time. CONCLUSION: The automatic contour propagation enables the fast and reproducible generation of a 4D model for the functional analysis of the heart. The interaction time is decreased from approx. 60 minutes to 10 minutes per case. Inter- and intraobserver differences of the extracted parameters are decreased significantly. PMID- 19283324 TI - Relationship between childhood underweight and dental crowding in deciduous teething. AB - OBJECTIVES: To investigate the relationship between low anthropometric indices and dental crowding in deciduous teething. METHODS: We assessed 794 children from 3 to 5 years old, both boys and girls, enrolled in public preschools in the city of Sao Luis, state of Maranhao, Brazil. Single-stage cluster sampling was used. This is a cross-sectional study, including the administration of a questionnaire, evaluation of dental occlusion and anthropometric assessment. Dental crowding, which is characterized by lack of space for the eruption of teeth in the line of the alveolar crest, is the dependent variable. Weight-for-age (W/A) and height for-age (H/A) indices were used for anthropometric assessments. Multivariate logistic regression analysis and descriptive analysis of the variables were used. Associations were estimated using odds ratio (OR), and the statistical inference was based on confidence intervals (95%CI). Type I error was set at 5% (alpha = 5%). RESULTS: The association between low anthropometric measures and dental crowding was statistically significant only when considering the weight-for-age index and among those children who did not use pacifiers. We found 2.19 children with dental crowding and malnutrition for each children without dental crowding and with malnutrition (OR = 2.19; 95%CI 1.19-4.04). CONCLUSION: This study suggests that low anthropometric measures are associated with dental crowding in deciduous teething among children who do not use pacifiers. However, further studies are needed to investigate possible causal relations between malnutrition and dental crowding. PMID- 19283325 TI - Pediatric tracheostomy. AB - OBJECTIVE: To provide an up-to-date review of pediatric tracheostomy, primarily focusing on indications, surgical technique, complications and hospital and home care. SOURCES: MEDLINE and PubMed databases were searched using the following keywords: tracheostomy, tracheotomy, children, newborn. SUMMARY OF THE FINDINGS: Indications for tracheostomy in children are changing. Today the most common indication is prolonged ventilation. The age at the time of the procedure has also changed, with a peak incidence of tracheostomy in patients less than 1 year old. Except under emergency conditions, pediatric tracheostomy should be performed in the operating room with the child intubated. A horizontal skin incision with vertical tracheal incision and no tracheal resection is recommended. Although post-tracheostomy complications are not uncommon, they usually do not need special treatment or surgical procedures. Tracheostomy mortality can occur in up to 40% of pediatric cases, however the tracheostomy related mortality rate is only 0 to 6%. CONCLUSIONS: The decision to perform a tracheostomy remains complex, and depends on several factors. The procedure is safe and with a low number of complications if carried out at a tertiary hospital by a trained and experienced team. PMID- 19283326 TI - Trace metals in oysters, Crassotrea sps., from UNESCO protected natural reserve of Urdaibai: space-time observations and source identification. AB - The seasonal and spatial variability of trace metal concentrations of oyster tissues (Crassostea sps.) was studied. The samples were collected between 2002 and 2004 along the Urdaibai estuary (Murueta, Kanala and Arteaga). The seasonal influence on some metal concentrations (higher values in spring-summer season than in autumn-winter season) is more pronounced than the local site-specific influence. Furthermore, within each location a significant time-specific dependence on the trace metal concentration variations can be detected. Substantially higher concentrations of Zn (445.5-2,403.3 mg kg(-1)) and Mn (29.4 169.6 mg kg(-1)) were recorded in oysters inhabiting estuarine locations, possibly indicating a markedly higher natural bioavailability at these locations. Principal component analysis demonstrated that oysters could be separated depending on the inhabiting areas: Sn for Murueta, essential metals for Kanala, and Co and Cd for Arteaga sampling points, respectively. PMID- 19283327 TI - Evaluation of a methodology for toxicity testing of volatile chlorinated hydrocarbons on marine organisms. AB - This study evaluated the suitability of sealed containers for toxicity testing to prevent loss of volatile chlorinated hydrocarbons (VCHs) with a range of Australian marine organisms including: micro-algae; sea urchin and oyster larvae in 44 mL sealed vials and fish larvae; amphipods; and juvenile polychaetes in 1 L sealed jars. Vials prevented volatilisation of VCHs during testing. Jars were less effective, with average losses of 46%. Growth and development of algae, sea urchins and oysters in vials was acceptable, indicating suitability of the methodology. Jars were suitable for amphipods and polychaetes; however, further evaluation of the fish test is required. PMID- 19283328 TI - Effects of low-magnitude, high-frequency mechanical stimulation in the rat osteopenia model. AB - SUMMARY: In this study, short-term, whole-body vertical vibration at 90 Hz improved trabecular bone quality. There was an improvement of bone quality and density in both osteoporotic and control rats. This treatment may therefore be an attractive option for the treatment of osteoporosis. INTRODUCTION: Aside from pharmacological treatment options, physical exercise is known to augment bone mass. In this study, the effects of whole-body vertical vibration (WBVV) on bone quality and density were evaluated using an osteoporotic rat model. METHODS: Sixty female Sprague Dawley rats were ovariectomized (C) or sham (SHAM) operated at the age of 3 months. After 3 months, both groups were divided into two subgroups that received either WBVV at 90 Hz for 35 days or no treatment. After sacrificing the rats, we evaluated vertebral bone strength, histomorphometric parameters, and bone mineral density (BMD). RESULTS: Treatment with WBVV resulted in improved biomechanical properties. The yield load after WBVV was significantly enhanced. According to yield load and Young's modulus, the treated OVX rats reached the level of the untreated SHAM animals. In all measured histomorphometric parameters, WBVV significantly improved bone density. Treatment with WBVV demonstrated greater effects on the trabecular bone compared to the cortical bone. The ash-BMD index showed significant differences between treated and untreated rats. CONCLUSION: Using WBVV as a non-pharmacological supportive treatment option for osteoporosis demonstrated an enhancement of bone strength and bone mass. This procedure may be an attractive option for the treatment of osteoporosis. PMID- 19283329 TI - High frequency of lipoprotein risk levels for cardiovascular disease in Takayasu arteritis. AB - The objective of this study is to characterize the lipoprotein risk levels in Takayasu arteritis (TA) patients and its possible association with disease activity and glucocorticoid use. Twenty-five female TA patients were consecutively included and compared with 30 age-, gender-, and body mass index matched healthy controls. Demographic features and the lipid profile were determined and cardiovascular risk levels were evaluated according to NCEP/ATPIII. Total cholesterol (TC), LDL-c, HDL-c, and triglycerides were determined after a 12-h overnight fast. Exclusion criteria were conditions that interfere in the lipid profile. The disease duration was 6.6 +/- 7.4 years; 30% had clinical activity and 80% had laboratory activity. Regarding NCEP/ATPIII risk levels, TA patients presented higher frequency of lipid risk compared to controls: high TC (48% vs. 20%, p = 0.04), low HDL-c (20% vs. 0%, p = 0.015), and high triglycerides (36% vs. 10%, p = 0.026). No difference was observed related to LDL-c risk levels between both groups (40% vs. 20%, p = 0.14). Remarkably, 60% of the patients had at least one lipid risk factor for cardiovascular disease. No difference in the lipids was observed between patients with and without clinical activity; however, those with laboratory activity showed lower levels of HDL-c (1.37 +/- 0.42 vs. 2.00 +/- 0.63 mmol/L, p = 0.012) than patients without this activity. A negative correlation was found between HDL-c and CRP levels (r = 0.42, p = 0.04). Lipids were similar in patients under glucocorticoid compared to those without this therapy. This is the first study to identify that TA, an inflammatory disease, has a proatherogenic lipid profile which is associated to laboratory disease activity. PMID- 19283330 TI - Mean platelet volume in children with familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is the most common inherited periodic fever syndrome characterized by recurrent episodes of serositis. Recently, a few studies have suggested that FMF is related to increased risk of atherosclerosis. Mean platelet volume (MPV) is a marker of platelet activation. Larger platelets are associated with increased atherosclerosis risk. The aim of the study is to evaluate levels of MPV in pediatric FMF patients during and between attacks. The study consisted of 48 patients during an attack (group 1), 63 patients in attack free period (at least 2 weeks after an attack, group 2), and 49 healthy controls (group 3). Erythrocyte sedimentation rate, C-reactive protein, white blood cell count, platelet count (PLT), and MPV levels were retrospectively recorded from the computerized patient database. Mean platelet volume was significantly lower in FMF patients during attack than in attack-free period (p = 0.00); however, there was no difference among attack-free patients and healthy controls (p = 0.38). The mean platelet counts of FMF patients during attack were higher than the healthy controls (p = 0.02). There was an inverse correlation between MPV and mean PLT in the attack-free period (r = -446, p = 0.01). This study suggests that an early atherosclerosis marker, MPV, is not elevated in pediatric FMF patients on colchicine treatment. PMID- 19283331 TI - Anti-dsDNA antibodies in Brazilian patients of mainly African descent with systemic lupus erythematosus: lack of association with lupus nephritis. AB - Renal disease is associated with morbidity and mortality in systemic lupus erythematosus (SLE) and anti-dsDNA antibodies with SLE immunopathogenesis. We investigated the dsDNA antibody profile of 84 Brazilian SLE patients, 27 with lupus nephritis. Thirty-six (39.1%) patients had dsDNA IgG antibodies shown in enzyme-linked immunosorbent assay (454.7 +/- 281.1 WHO units/mL), nine presenting renal disease. The following profile of dsDNA antibodies was demonstrated in Crithidia luciliae test: IgA (seven out of 36; 19.4%), IgG (22 out of 36, 66.1%); IgM (nine out of 36, 25.0%), and IgE (four out of 36, 11.1%). Two or three isotypes of dsDNA antibodies were observed in nine (25.0%) patients, while 11 (30.5%) were seronegative in the C. luciliae test. Patients with dsDNA antibodies had lower serum C3 and C4 when compared with SLE individuals without these immunoglobulins (P < 0.01 and P < 0.001, respectively). There was no association between any dsDNA antibody isotype and lupus kidney disease nor was anti-dsDNA IgM antibody associated with absence of nephritis. PMID- 19283332 TI - Prevalence of ventricular discordance and its relation to functional capacity in idiopathic dilated cardiomyopathy. AB - INTRODUCTION: Although left ventricular (LV) dilatation is the most distinguishing morphologic feature of idiopathic dilated cardiomyopathy (IDC), right ventricular (RV) dilatation may variably contribute to total cardiac enlargement. The prevalence and functional importance of the relative degree of left and right ventricular dilatation has not been comprehensively studied using cardiac magnetic resonance imaging (MRI). METHODS: Our prospective study included 58 consecutive IDC patients with a LV ejection fraction <40% and NYHA functional class > or =2. MRI was performed with a 1.5 Tesla scanner for RV and LV dimensional and functional analysis. Cardiopulmonary exercise testing was used for evaluation of exercise capacity. Patients were grouped into tertiles based on the distribution of LV end-diastolic volume. RESULTS: Compared to control subjects a considerable heterogeneity in the relative degree of left and right ventricular dilatation was noted in IDC patients. Within the entire patient group, a strong correlation between the degree of ventricular volume discordance and the extent of LV enlargement was observed (r = 0.8; P < 0.001). Tertile analysis revealed that the LV/RV volume ratio significantly differed in the three subgroups of patients (1.0 +/- 0.3 vs. 1.5 +/- 0.4 vs. 2.1 +/- 0.9; P < 0.001). Only weak correlations between MRI data and patients' functional capacity were found. LV ejection fraction was identified as the only independent predictor of maximum oxygen consumption in our setting. CONCLUSION: In IDC patients the degree of ventricular volume discordance is strongly related to the extent of left ventricular enlargement. However, comprehensive biventricular assessment of cardiac function by MRI adds little to our understanding of the cardiac mechanisms limiting exercise tolerance when compared to exclusive left ventricular measurements. PMID- 19283333 TI - Perioperative antiplatelet therapy in patients at risk for coronary stent thrombosis undergoing noncardiac surgery. PMID- 19283334 TI - Catheter ablation of atrial fibrillation using the Navx-/Ensite-system and a CT /MRI-guided approach. AB - BACKGROUND: Catheter ablation has become the first line of therapy in patients with symptomatic, recurrent, drug refractory atrial fibrillation. However, catheter ablation of atrial fibrillation is still a challenge. This is partially due to the high degree of variability with regard to the individual anatomy. Nevertheless, 3D imaging systems (CT, MRI) provide detailed information about the individual left atrial and pulmonary vein morphology. A 3D CT or MRI reconstruction of the left atrium can be displayed in the Navx-/Ensite-system in a synchronised way during the ablation procedure, thereby facilitating the intervention. This study summarizes our preliminary experience with different strategies of AF ablation using the Navx-/Ensite-system and a CT-/MRI-guided approach. METHODS: In a total of 41 patients, cardiac MRI (n = 7) or multi detector spiral computed tomography (n = 34) was performed prior to an ablation procedure. Catheter ablation was performed for paroxysmal atrial fibrillation in 31 patients and for persistent atrial fibrillation in 10 patients. A 3D MRI or high resolution spiral CT data acquisition was performed and a surface rendered model of the LA was created. This model was displayed in the Navx-/Ensite-system throughout the ablation procedure. RESULTS: Catheter ablation was performed using the Navx-system (n = 38) or the Ensite-system (n = 3). Three strategies were used depending on the type of atrial fibrillation: segmental isolation of the pulmonary veins (facilitated by a 3D real-time visualization of the ablation catheter and a circumferential mapping catheter; group A: 20 patients), linear lesions (group C: 3 patients) and a combined approach (group B; 18 patients). The CT-/MRI-models provided an excellent overview over the pulmonary veins and the left atrial appendage. They revealed a high degree of variability with regard to the individual anatomy (e.g. dimensions of the left atrial appendage, pulmonary vein ostia). The CT scans provided a more detailed reconstruction of the left atrial anatomy than the MRI scans (especially in patients who were in atrial fibrillation at the time of the data acquisition). In some patients, the CT-/MRI models revealed a very small diameter of some pulmonary veins or side branches close to the ostium (e.g. right inferior pulmonary vein). Therefore, no attempt was made to achieve complete pulmonary vein isolation in some patients. In group A, 16/20 (80%) patients had no arrhythmia recurrence [mean follow-up 359 days (SD +/- 317 days)]. Twelve out of eighteen (67%) patients in group B [mean follow-up 452 days (SD +/- 311 days)] and 2/3 (67%) patients in group C did not experience an arrhythmia recurrence [mean follow-up 1,000 days (SD +/- 34 days)]. There were no major complications. CONCLUSIONS: The information derived from 3D CT- or MRI reconstructions facilitates AF ablations performed with the Navx-/Ensite-mapping system and enhances the safety of these procedures. Furthermore, the availability of an additional impedance-based 3D real-time visualization of the ablation catheter and the circular mapping catheter placed in the pulmonary veins represents a major advantage of the Navx system. PMID- 19283335 TI - Hypogenesis of intramural vascularity and perivascular plexuses of gallbladder in patients with congenital biliary dilatation. AB - BACKGROUND: We hypothesized neuronal disorders of the biliary tract as the cause of congenital biliary dilation (CBD). METHODS: Gallbladders were removed from a total of 15 patients who were categorized into two study groups: a CBD group (eight patients) and in a control group (neuroblastoma, acute myelogenous leukemia, wandering gallbladder, Wilms' tumor, glycogen storage disease, familial amyloid polyneuropathy; seven patients). Whole-mount preparations of the gallbladders were made to immunostain the intramural nerves. RESULTS: The intramural vascularity in the gallbladders of the CBD group (5.5 +/- 1.9/cm(2)) was significantly lower than that in the control group (27.6 +/- 14.4/cm(2)). The rate of perivascular plexuses on the surface of intramural vessels was also significantly lower in the CBD group than in the controls (37.7 +/- 18.1 vs. 80.2 +/- 17.4%, respectively). The numbers of ganglion cells per visual field were 38.5 +/- 24.0 and 42.3 +/- 20.6, respectively, in the CBD and control groups; this difference was not statistically significant. CONCLUSIONS: There may be a mechanism in CBD causing contractile failure and dilatation of the biliary tract as a result of decreased intramural blood flow that accompanies the diminished vascularity. PMID- 19283336 TI - Improvement of early delayed gastric emptying in patients with Billroth I type of reconstruction after pylorus preserving pancreatoduodenectomy. AB - BACKGROUND: Early delayed gastric emptying (DGE) is the most common complication after pylorus-preserving pancreatoduodenectomy (PpPD). Recently, a vertical antecolic reconstruction for duodenojejunostomy was recommended to decrease the incidence of early DGE in patients with Billroth II-type reconstruction after PpPD. However, Billroth I-type reconstruction (B-I) after PpPD is still favored in Japan. METHODS: Twelve consecutive patients with B-I were prospectively enrolled. Our technique includes an end-to-side duodenojejunostomy and alignment of the stomach contours with fixation of the greater omentum to the abdominal wall in order to promote passage from the stomach through the jejunal loop. DGE was evaluated according to the consensus definition of the International Study Group of Pancreatic Surgery (ISGPS). RESULTS: DGE was absent, with the nasogastric tube removed within 3 days in all patients. Mean duration of nasogastric tube placement was 1.5 +/- 0.4 days. Mean maximum suction volume was 85 +/- 32 ml/day. CONCLUSION: Preliminary results were encouraging simply with relief of the outflow disturbance around the duodenojejunostomy in patients with B-I after PpPD. These findings warrant further prospective randomized trials at either multiple or high-volume centers. PMID- 19283337 TI - Bone mass in schoolchildren in Brazil: the effect of racial miscegenation, pubertal stage, and socioeconomic differences. AB - The purpose of this study was to evaluate bone mass by phalanges ultrasound in healthy white and black schoolchildren in relationship to socioeconomic level, pubertal stage, and body composition. Included were 1,356 healthy schoolchildren aged from 6 to 11 years from different socioeconomic levels and both genders; all were placed into white and black groups. Weight, height, body mass index, fat percentage, fat mass, and lean mass were evaluated by anthropometric methods, and AD-SoS bone quantity and UBPI bone quality were evaluated using a third generation IGEA phalanges DBM Sonic BP ultrasound. Data were compared using the Mann-Whitney, chi-squared, correlation coefficient, and analyses of multiple linear regression statistical tests with 5% significance. Black schoolchildren predominated in the low socioeconomic levels. Higher values of weight and height for black boys and girls were observed in the lean mass in relation to white children of the same gender and age. An increasing variation in the bone quantity mean was observed from 6 to 11 years of age and with pubertal stage for both genders and skin color. The white schoolchildren presented higher values of bone quantity and quality in relation to the black children. The anthropometric, gender, and socioeconomic level variables explained only 16 and 11% of the variability of bone quantity and quality, respectively. As such, the present study, carried out with healthy black and white Brazilian schoolchildren, demonstrated higher bone mass, as evaluated by ultrasound, in white than in black schoolchildren. PMID- 19283338 TI - Genetic diversity of porcine Norovirus and Sapovirus: Canada, 2005-2007. AB - Noroviruses and sapoviruses are members of the family Caliciviridae and emerging enteric pathogens of humans and animals. Since their discovery and characterization in swine, relatively few strains have been described in detail. In order to investigate their genetic diversity, a total of 266 fecal samples collected in the province of Quebec, Canada, between 2005 and 2007 were screened for the presence of caliciviruses by RT-PCR using broadly reactive primers. Genetically heterogeneous caliciviruses were detected on the majority of farms. Typical noroviruses related to known swine genotypes were present on 20% of the farms. Sapoviruses were detected on 75% of the farms and were the most heterogeneous group. Further characterization of selected strains in their 3' end parts was carried out for their classification and unveiled possibly new clusters of sapoviruses. No human-like noroviruses or sapoviruses were detected in the present study. PMID- 19283339 TI - Design of compound libraries for fragment screening. AB - Approaches to the design of libraries for fragment screening are illustrated with reference to a 20 k generic fragment screening library and a 1.2 k generic NMR screening library. Tools and methods for library design that have been developed within AstraZeneca are described, including Foyfi fingerprints and the Flush program for neighborhood characterization. It will be shown how Flush and the BigPicker, which selects maximally diverse sets of compounds, are used to apply the Core and Layer method for library design. Approaches to partitioning libraries into cocktails are also described. PMID- 19283340 TI - Chromium improves glucose uptake and metabolism through upregulating the mRNA levels of IR, GLUT4, GS, and UCP3 in skeletal muscle cells. AB - The aim of this study was to evaluate the impact of three different chromium forms as chromic chloride (CrCl), chromium picolinate (CrPic), and a newly synthesized complex of chromium chelated with small peptides (CrSP) on glucose uptake and metabolism in vitro. In cultured skeletal muscle cells, chromium augmented insulin-stimulated glucose uptake and metabolism as assessed by a reduced glucose concentration of culture medium. At the molecular level, insulin significantly increased the mRNA levels of insulin receptor (IR), glucose transporter 4 (GLUT4), glycogen synthase (GS), and uncoupling protein-3 (UCP3), and these impacts can be enhanced by the addition of chromium, especially in the form of CrSP. Collectively, results of this study demonstrate that chromium improves glucose uptake and metabolism through upregulating the mRNA levels of IR, GLUT4, GS, and UCP3 in skeletal muscle cells, and CrSP has higher efficacy on glucose uptake and metabolism compared to the forms of CrCl and CrPic. PMID- 19283341 TI - Hypoglycemic activity of Grifola frondosa rich in vanadium. AB - The hypoglycemic activity of fermented mushroom of Grifola frondosa rich in vanadium (GFRV) was studied in this paper. Alloxan- and adrenalin-induced hyperglycemic mice were used in the study. The blood glucose and the HbA1c of the mice were analyzed respectively. After the mice were administered (ig) with GFRV, the blood glucose and the HbA1c of alloxan-induced hyperglycemic mice decreased (p < 0.05, p < 0.01) and ascension of blood glucose induced by adrenalin was inhibited (p < 0.01). Also, the bodyweight of the alloxan-induced hyperglycemic mice was increased gradually. In the fermented mushroom of G. frondosa, vanadium at lower doses in combination with G. frondosa induced significant decreases of the blood glucose and HbA1c levels in hyperglycemic mice. PMID- 19283342 TI - Comparison of hypoglycemic activity of trace elements absorbed in fermented mushroom of Coprinus comatus. AB - The effect of fermented mushroom of Coprinus comatus rich in trace elements, including vanadium, chromium, zinc, magnesium, copper, iron, and nickel, on glycemic metabolism was studied in this paper. Alloxan-induced hyperglycemic mice were used in the study. The blood glucose, glycohemoglobin, and glycogen synthesis of the mice were analyzed, respectively. At the same time, the gluconeogenesis of the normal mice was also determined. After the mice were administered (ig) with C. comatus rich in vanadium (CCRV), the blood glucose and the glycohemoglobin of alloxan-induced hyperglycemic mice decreased (p < 0.05, p < 0.01), glycogen synthesis of alloxan-induced hyperglycemic mice elevated (p < 0.01), the gluconeogenesis of the normal mice was inhibited (p < 0.01), and the sugar tolerance of the normal mice was improved. However, the same result did not occur in other groups. Vanadium at lower doses in combination with C. comatus induced significant effect on glycemic metabolism in mice. PMID- 19283343 TI - Introduction. Idebenone in the treatment of Friedreich ataxia. PMID- 19283344 TI - Friedreich ataxia: the clinical picture. AB - Friedreich ataxia (FRDA) is a rare autosomal recessive hereditary disorder that affects approximately 1 in 50,000 Caucasians. It is caused by hyperexpansion of GAA repeats in the first intron of the frataxin gene. Initial symptoms of FRDA usually appear around the beginning of the second decade of life. In addition to neuropathological disabilities such as ataxia, sensory loss, and muscle weakness, common signs are scoliosis, foot deformity, and hypertrophic cardiomyopathy. Approximately 10 % of patients with FRDA develop diabetes. The neuronopathy in the dorsal root ganglia, accompanied by the loss of peripheral sensory nerve fibres and the degeneration of posterior columns of the spinal cord, is a hallmark of the disease and is responsible for the typical combination of signs and symptoms specific to FRDA. Variation in neurophysiological abnormalities is correlated with the size of the GAA repeat expansion and likely accounts for individual variation in the progression of FRDA. Despite a range of disease severity, most patients will lose their ability to walk, stand, or sit without support within 10 to 15 years of disease onset. In addition to a review of the clinicopathological features of FRDA, a discussion of recent advances in our understanding of the underlying molecular mechanisms is provided. PMID- 19283345 TI - The pathogenesis of Friedreich ataxia and the structure and function of frataxin. AB - Understanding the role of frataxin in mitochondria is key to an understanding of the pathogenesis of Friedreich ataxia. Frataxins are small essential proteins whose deficiency causes a range of metabolic disturbances, which include oxidative stress, deficit of iron-sulphur clusters, and defects in heme synthesis, sulfur amino acid and energy metabolism, stress response, and mitochondrial function. Structural studies carried out on different orthologues have shown that the frataxin fold consists of a flexible N-terminal region present only in eukaryotes and in a highly conserved C-terminal globular domain. Frataxins bind iron directly but with very unusual properties: iron coordination is achieved solely by glutamates and aspartates exposed on the protein surface. It has been suggested that frataxin function is that of a ferritin-like protein, an iron chaperone of the iron sulphur cluster machinery and heme metabolism and/or a controller of cellular oxidative stress. To understand FRDA pathogenesis and to design novel therapeutic strategies, we must first precisely identify the cellular role of frataxin. PMID- 19283346 TI - Multicellular models of Friedreich ataxia. AB - Patients with Friedreich ataxia (FRDA) have severely reduced levels of the mitochondrial protein frataxin, which results from a large GAA triplet-repeat expansion within the frataxin gene (FXN). High evolutionary conservation of frataxin across species has enabled the development of disease models of FRDA in various unicellular and multicellular organisms. Mouse models include classical knockout models, in which the Fxn gene is constitutively inactivated, and knock in models, in which a GAA repeat mutation or the conditional allele is inserted into the genome. Recently, "humanised" GAA repeat expansion mouse models were obtained by combining the constitutive knockout with the transgenic expression of a yeast artificial chromosome carrying the human FRDA locus. In lower organisms such as Caenorhabditis elegans and Drosophila, straight-forward and conditional RNA interference technology has provided an easy way to knock down frataxin expression. Conditional mouse models have been used for pre-clinical trials of potential therapeutic agents, including idebenone, MnTBAP (a superoxide dismutase mimetic), and iron chelators. Various models of FRDA have shown that different, even opposite, phenotypes can be observed, depending on the level of frataxin expression. Additional studies with animal models will be essential for an enhanced understanding of the disease pathophysiology and for the development of better therapies. PMID- 19283347 TI - Idebenone: an emerging therapy for Friedreich ataxia. AB - This paper reviews the history and pre-clinical development of idebenone and summarises the results of clinical studies, published from 1999 to 2008, on the use of idebenone in the treatment of patients with Friedreich ataxia (FRDA). As a benzoquinone that can undergo reversible redox reactions, idebenone influences the electron balance in mitochondria. In vitro studies have shown that it acts both as an anti-oxidant, preventing damage to the mitochondrial membrane, and, more importantly, as an electron carrier, supporting mitochondrial function and adenosine triphosphate (ATP) production. In clinical studies, idebenone has been well tolerated by patients with various pathological conditions. The most common adverse events have been gastrointestinal effects of mild to moderate severity. No neurotoxic or adverse cardiac reactions have been reported in pre-clinical or clinical studies. The good safety profile of idebenone is supported by large clinical trials in Alzheimer's disease and by post-marketing surveillance. Phase 1 studies demonstrated the safety and tolerability of idebenone at relatively high doses (up to 60 mg/kg/day). Results from 11 clinical studies (randomised, controlled, and open-label trials), involving a total of about 200 patients, provide evidence of improvement in both cardiac hypertrophy and neurological symptoms among patients with FRDA treated with idebenone. PMID- 19283348 TI - Pharmacokinetic properties and metabolism of idebenone. AB - Four phase 1 studies were conducted to assess the pharmacokinetics and metabolism of idebenone (including parent drug and inactive metabolites QS10, QS6, and QS4) and to evaluate the safety of a wide range of idebenone doses and regimens in healthy adult men. After a single oral dose of idebenone 150 mg, 750 mg, or 1050 mg in fasted or fed subjects, blood samples were taken for up to 72 hours. In one study, after a single oral dose and a 7-day washout period, subjects received repeated doses of idebenone 150 mg or 750 mg every 8 hours for 14 days. In the repeated-dose study, urine samples also were taken. Plasma and urine samples were analysed with the use of liquid chromatography with tandem mass spectrometry. Non compartmental standard pharmacokinetic methods were used. In these studies, a total of 69 subjects ranging in age from 19 to 41 years (body weight, 57-94.6 kg) were included. Plasma concentrations of parent idebenone were low but increased in proportion to dose and increased approximately five-fold in the presence of food. Total QS4 was the main metabolite in plasma and urine. The most common adverse events were loose stool, fatigue, headache, and disturbances in attention. Idebenone was well tolerated in single oral doses up to 1050 mg and in repeated daily doses up to 2250 mg. Idebenone showed linear pharmacokinetics after single and repeated oral dosing. Administration after a meal resulted in the highest exposure to parent idebenone. PMID- 19283349 TI - Evaluating the progression of Friedreich ataxia and its treatment. AB - Friedreich ataxia is characterised by slowly progressive neurodegeneration and cardiomyopathy. Currently, no treatments have been proven to delay, prevent, or reverse the inexorable decline that occurs in this condition; however, several pharmaceutical agents are undergoing clinical assessment. Because initial beneficial therapies are likely to slow disease progression rather than reverse morbidity, the need for accurate measurement tools that will detect such subtle benefits is critical. The impact of Friedreich ataxia on the nervous system has been assessed largely through the use of rating scales and functional composite measures, and a number of patient reported outcome measures in Friedreich ataxia have been studied. However, on the basis of published reports on the performance of these measures, none clearly stands out as the best for use in clinical trials. Refinement of existing tools and development of new tools will be needed to maximise the chance of detecting small but clinically significant benefits of therapeutic agents in patients with Friedreich ataxia. PMID- 19283350 TI - Clinical experience with high-dose idebenone in Friedreich ataxia. AB - Several reports in the literature describe the effects of low-dose (5 mg/kg/day) idebenone in significantly reducing cardiac hypertrophy in patients with Friedreich ataxia. However, the effects of idebenone on neurological function have not been reliably determined in these studies; when neurological parameters were reported, results were often inconclusive, usually because of subject heterogeneity and lack of adequate statistical power. In two of these studies, some patients showed beneficial effects of idebenone on their cardiomyopathy only when the dose was increased, prompting the systematic investigation of higher doses of idebenone. Following a phase 1 dose escalation study, a phase 2 tolerability and efficacy trial with low, intermediate, and high doses of idebenone was conducted. The results suggested that treatment with intermediate- and high-dose idebenone had beneficial effects on neurological symptoms. On the basis of these results, two phase 3 trials have been initiated, one in the United States with young ambulatory patients and one in Europe without limits on age and disease severity. PMID- 19283352 TI - [Diabetes mellitus - perioperative management]. AB - The prevalence of diabetes in hospitalized adults is conservatively estimated at 12-25% and rising. Poor glucose control and presence of diabetes complications (e.g. diabetic nephropathy, diabetic neuropathy, atherosclerosis) are commonly regarded as risk factors for perioperative morbidity and mortality. Thus it is crucial to determine diabetes comorbidities preoperatively in order to avoid perioperative renal and cardiovascular complications. Perioperative glycemic control is challenging due to preoperative changes in diabetes treatment and the effects of surgery-associated stress hyperglycemia. For patients in general surgical units, evidence for specific glycemic goals is based on epidemiologic and physiologic data rather than clinical trials. According to guidelines of the German Society of Nutrition, the approximation of normoglycemia is reasonable as long as hypoglycemia is avoided (suggested range for plasma glucose 80-145 mg/dL). PMID- 19283353 TI - A phase I study of palliative chemoradiation therapy with paclitaxel and cisplatin for local symptoms due to an unresectable primary advanced or locally recurrent gastric adenocarcinoma. AB - PURPOSE: To establish the maximum tolerated dose and dose-limiting toxicity of chemoradiation with paclitaxel (PTX) and cisplatin (CDDP) for patients with local symptoms due to unresectable primary advanced or locally recurrent gastric adenocarcinoma located at left-upper abdomen. METHODS: Chemotherapy consisted of PTX at escalating doses of 40-80 mg/m(2) per day and CDDP at escalating doses of 20-25 mg/m(2) per day on days 1, 15, and 29. Concurrent radiation was administered up to a dose of 45 Gy for 5 weeks. RESULTS: A total of nine patients were enrolled, of which six were into level 1 (PTX 60 mg/m(2) and CDDP 20 mg/m(2)) and three into level -1 (PTX 50 mg/m(2) and CDDP 20 mg/m(2)). At level 1, one patient developed grade 3 fatigue, and the other experienced grade 5 DIC, grade 5 pneumonia, grade 4 thrombocytopenia, grade 3 hyponatremia, and grade 3 esophagitis as dose-limiting toxicities. A palliative effect was observed in eight of nine patients; six of six patients at level 1 and two of three at level 1. CONCLUSION: PTX 50 mg/m(2) and CDDP 20 mg/m(2) given biweekly with concurrent radiation therapy of 45 Gy were well tolerated. PMID- 19283354 TI - Delivery of high levels of anti-proliferative nucleoside triphosphates to CYP3A expressing cells as a potential treatment for hepatocellular carcinoma. AB - PURPOSE: Hepatocellular carcinoma (HCC) is a life-threatening condition with only one drug treatment regimen approved for use. Oncolytic nucleosides are minimally effective against HCC putatively because of their inability to achieve cytotoxic levels of the active metabolite [nucleoside triphosphate (NTP)] in tumor cells at doses that are well tolerated. The aim of our studies was to explore the utility of CYP3A-activated prodrugs of cytarabine and fludarabine monophosphate for the treatment of HCC. METHODS: Prodrugs of cytarabine and fludarabine monophosphates were evaluated for their ability to safely achieve NTP levels in the liver of normal mice that are cytotoxic to hepatoma cells. RESULTS: While therapeutic levels of NTPs are achieved in the livers of normal rodents after administration of the prodrugs, only MB07133 achieved these levels without exhibiting signs of liver toxicity or myelosuppression. CONCLUSIONS: As the levels of araCTP achieved in the liver at therapeutic doses are only toxic to proliferating cells (such as those in HCC tumors), but not the non-proliferative adjacent tissue, MB07133 treatment has the potential to be both efficacious and well tolerated in HCC patients. PMID- 19283355 TI - [Transoral vertebroplasty: an alternative for operative treatment of metastases of the upper cervical spine]. AB - Analogue to the demographic changes and the accompanying increased incidence of tumorous diseases, the number of patients with metastatic bone tumors of the spine is also increasing. Metastatic bone tumors are the most significant cause of pain in cancer patients. Pain and instability are the main indications for surgery. Minimally invasive procedures are recommended in patients with a poor medical condition and with a poor prognosis.Transoral vertebroplasty can be successfully used to reduce pain and provide stability in the palliative treatment of metastases of the vertebral axis. This procedure has the advantage of providing rapid pain relief and spinal stabilization.The operative technique is described and discussed with reference to the current literature. As an example the case of a 67-year-old patient is described, who was suffering from prostate cancer and a painful metastasis of the dens axis. After interdisciplinary consensus, transoral vertebroplasty was performed. The procedure was effective in achieving pain relief and providing stability and 7 months after the operation no further spinal metastases had occurred. PMID- 19283356 TI - [Ankle osteosynthesis infection: vacuum therapy as the treatment of choice]. AB - BACKGROUND: Should osteosynthesis infection occur after ankle fractures in patients with microangiopathy, the infection needs to be cured quickly and safely to protect implants, bone, and tendons. Vacuum therapy (TNP) provides a perfect treatment plan that keeps the inpatient time low whilst ensuring high patient comfort. METHODS AND PATIENTS: Four angiopathic patients with deep wound infection after ankle osteosynthesis were treated. At admission, loss of stability and spread of the infection were the immediate risks. To prevent this, we treated the patients with vacuum therapy after initial debridement until skin cover was achieved. RESULTS: In all cases stability was secured and after 2 dressing changes, the swabs were sterile. The inpatient time was 13 days; overall vacuum therapy time was 59 days. Patient satisfaction was high. DISCUSSION: Vacuum therapy is a perfect strategy after surgical debridement and before secondary mesh grafting. It protects the stability of the bone and open-lying delicate structures in patients with infected osteosynthesis of the ankle and angiopathy and offers an easy, safe, and successful treatment path with a short inpatient time. PMID- 19283357 TI - Magnetic properties of the microorganism Candidatus Magnetoglobus multicellularis. AB - Magnetotactic microorganisms use the interaction of internal biomineralized nanoparticles with the geomagnetic field to orientate. The movement of the magnetotactic multicellular prokaryote Candidatus Magnetoglobus multicellularis under an applied magnetic field was observed. A method using digital image processing techniques was used to track the organism trajectory to simultaneously obtain its body radius, velocity, U-turn diameter, and the reorientation time. The magnetic moment was calculated using a self-consistent method. The distribution of magnetic moments and radii present two well-characterized peaks at (9 +/- 2) x 10(-15) and (20 +/- 3) x 10(-15) A m(2) and (3.6 +/- 0.1) and (4.3 +/- 0.1) mum, respectively. For the first time, simultaneous determination of the distribution of the organism radii and magnetic moment was obtained from the U turn method by a new digital imaging processing. The bimodal distributions support an organism reproduction process model based on electron microscopy observations. These results corroborate the prokaryote multicellular hypothesis for Candidatus M. multicellularis. PMID- 19283358 TI - [Vertebrobasilar occlusions : Pathophysiology, diagnostics and treatment]. AB - Acute vertebrobasilar occlusions (VBO) are dramatic clinical events with a mortality of up to 90% under standard medical treatment. If VBO is suspected a diagnosis of the vessel status has to be achieved immediately. For this purpose CT/CTA and MRI/MRA are equivalent diagnostic tools in the emergency setting. In contrast to the anterior circulation, local endovascular treatment is the established therapy for the posterior circulation as an underlying arteriosclerotic stenosis remains in 50% of the cases after intravenous fibrinolysis. Nevertheless, systemic fibrinolysis is considered the preferred option in cases where a neurointerventional center cannot be reached within a reasonable time frame and the patient can subsequently be transported for local therapy of a residual stenosis in order to prevent reocclusion ("drip and ship"). Profound clinical and pathophysiological knowledge is the absolute prerequisite for the correct application of state-of-the-art neurointerventional therapy. This review paper focuses on the clinical and pathophysiological details that are crucial for decision-making. PMID- 19283360 TI - Genetic control of wheat quality: interactions between chromosomal regions determining protein content and composition, dough rheology, and sponge and dough baking properties. AB - While the genetic control of wheat processing characteristics such as dough rheology is well understood, limited information is available concerning the genetic control of baking parameters, particularly sponge and dough (S&D) baking. In this study, a quantitative trait loci (QTL) analysis was performed using a population of doubled haploid lines derived from a cross between Australian cultivars Kukri x Janz grown at sites across different Australian wheat production zones (Queensland in 2001 and 2002 and Southern and Northern New South Wales in 2003) in order to examine the genetic control of protein content, protein expression, dough rheology and sponge and dough baking performance. The study highlighted the inconsistent genetic control of protein content across the test sites, with only two loci (3A and 7A) showing QTL at three of the five sites. Dough rheology QTL were highly consistent across the 5 sites, with major effects associated with the Glu-B1 and Glu-D1 loci. The Glu-D1 5 + 10 allele had consistent effects on S&D properties across sites; however, there was no evidence for a positive effect of the high dough strength Glu-B1-al allele at Glu-B1. A second locus on 5D had positive effects on S&D baking at three of five sites. This study demonstrated that dough rheology measurements were poor predictors of S&D quality. In the absence of robust predictive tests, high heritability values for S&D demonstrate that direct selection is the current best option for achieving genetic gain in this product category. PMID- 19283361 TI - The vascular actions of insulin control its delivery to muscle and regulate the rate-limiting step in skeletal muscle insulin action. AB - Evidence suggests that insulin delivery to skeletal muscle interstitium is the rate-limiting step in insulin-stimulated muscle glucose uptake and that this process is impaired by insulin resistance. In this review we examine the basis for the hypothesis that insulin acts on the vasculature at three discrete steps to enhance its own delivery to muscle: (1) relaxation of resistance vessels to increase total blood flow; (2) relaxation of pre-capillary arterioles to increase the microvascular exchange surface perfused within skeletal muscle (microvascular recruitment); and (3) the trans-endothelial transport (TET) of insulin. Insulin can relax resistance vessels and increase blood flow to skeletal muscle. However, there is controversy as to whether this occurs at physiological concentrations of, and exposure times to, insulin. The microvasculature is recruited more quickly and at lower insulin concentrations than are needed to increase total blood flow, a finding consistent with a physiological role for insulin in muscle insulin delivery. Microvascular recruitment is impaired by obesity, diabetes and nitric oxide synthase inhibitors. Insulin TET is a third potential site for regulating insulin delivery. This is underscored by the consistent finding that steady-state insulin concentrations in plasma are approximately twice those in muscle interstitium. Recent in vivo and in vitro findings suggest that insulin traverses the vascular endothelium via a trans-cellular, receptor-mediated pathway, and emerging data indicate that insulin acts on the endothelium to facilitate its own TET. Thus, muscle insulin delivery, which is rate-limiting for its metabolic action, is itself regulated by insulin at multiple steps. These findings highlight the need to further understand the role of the vascular actions of insulin in metabolic regulation. PMID- 19283362 TI - Prevention of neural tube defects by loss of function of inducible nitric oxide synthase in fetuses of a mouse model of streptozotocin-induced diabetes. AB - AIMS/HYPOTHESIS: Maternal diabetes during pregnancy increases the risk of congenital malformations such as neural tube defects (NTDs). Although the mechanism of this effect is uncertain, it is known that levels of nitric oxide synthase (NOS) and nitric oxide are elevated in embryos of a mouse model of diabetes. We postulated that overproduction of nitric oxide causes diabetes induced congenital malformations and that inhibition of inducible NOS (iNOS) might prevent diabetic embryopathy. METHODS: Mice were rendered hyperglycaemic by intraperitoneal injection of streptozotocin. The incidence of congenital malformations including NTDs was evaluated on gestational day 18.5. We assessed the involvement of iNOS in diabetes-induced malformation by administering ONO 1714, a specific inhibitor of iNOS, to pregnant mice with streptozotocin-induced diabetic mice and by screening mice with iNOS deficiency due to genetic knockout (iNos(-/-)). RESULTS: ONO-1714 markedly reduced the incidence of congenital anomalies, including NTDs, in fetuses of a mouse model of diabetes. It also prevented apoptosis in the head region of fetuses, indicating that iNOS is involved in diabetes-related congenital malformations. Indeed, no NTDs were observed in fetuses of diabetic iNos(-/-) mice and the incidence of other malformations was also markedly reduced. CONCLUSIONS/INTERPRETATION: We conclude that increased iNOS activity during organogenesis plays a crucial role in the pathogenesis of diabetes-induced malformations and suggest that inhibitors of iNOS might help prevent malformations, especially NTDs, in diabetic pregnancy. PMID- 19283363 TI - Amphetamine pretreatment facilitates appetitive sexual behaviors in the female rat. AB - RATIONALE: Intermittent treatment of rats with psychomotor stimulants induces behavioral sensitization to their motor-stimulating effects. This sensitization involves an increase in mesolimbic and nigrostriatal dopamine release, and in male rats, facilitates sexual behavior. OBJECTIVES: The aim of this study is to investigate the effect of repeated injections of D-amphetamine on appetitive and consummatory sexual behaviors in female rats. MATERIALS AND METHODS: Sexually experienced or naive females were injected with either D-amphetamine (1 mg/kg, i.p.) or saline every other day for three injections each. After each amphetamine injection, females were placed either in a bilevel testing chamber or in their home cages. After saline injections, females were placed in bilevel chambers. Following a 3-week washout period, females were tested for sexual behavior in bilevel chambers in a drug-free state. RESULTS: Amphetamine pre-exposure facilitated the display of solicitations, hops and darts, and female-male mounting (FMM), regardless of whether the drug was paired with the testing environment. CONCLUSION: Intermittent amphetamine pretreatment that induces behavioral sensitization facilitates appetitive sexual behaviors in female rats, as has been shown previously in male rats. This suggests that the physiological substrates that modulate sensitized responses to psychomotor stimulants also mediate sensitized appetitive responses to sexual cues, including solicitation, hops and darts, and FMM. As in male rats, this facilitation was a direct consequence of amphetamine sensitization and not due to conditioned associations between drug and test environment. PMID- 19283366 TI - Environmental monitoring of phenolic pollutants in water by cloud point extraction prior to micellar electrokinetic chromatography. AB - Many aromatic compounds can be found in the environment as a result of anthropogenic activities and some of them are highly toxic. The need to determine low concentrations of pollutants requires analytical methods with high sensitivity, selectivity, and resolution for application to soil, sediment, water, and other environmental samples. Complex sample preparation involving analyte isolation and enrichment is generally necessary before the final analysis. The present paper outlines a novel, simple, low-cost, and environmentally friendly method for the simultaneous determination of p nitrophenol (PNP), p-aminophenol (PAP), and hydroquinone (HQ) by micellar electrokinetic capillary chromatography after preconcentration by cloud point extraction. Enrichment factors of 180 to 200 were achieved. The limits of detection of the analytes for the preconcentration of 50-ml sample volume were 0.10 microg L(-1) for PNP, 0.20 microg L(-1) for PAP, and 0.16 microg L(-1) for HQ. The optimized procedure was applied to the determination of phenolic pollutants in natural waters from San Luis, Argentina. PMID- 19283364 TI - The sigma-1 antagonist BMY-14802 inhibits L-DOPA-induced abnormal involuntary movements by a WAY-100635-sensitive mechanism. AB - RATIONALE: Levodopa (L-DOPA), the gold standard treatment for Parkinson's disease (PD), eventually causes L-DOPA-induced dyskinesia (LID) in up to 80% of patients. In the 6-hydroxydopamine (6-OHDA) rat model of PD, L-DOPA induces a similar phenomenon, which has been termed abnormal involuntary movement (AIM). We previously demonstrated that BMY-14802 suppresses AIM expression in this model. OBJECTIVES: Although BMY-14802 is widely used as a sigma-1 antagonist, it is also an agonist at serotonin (5-HT) 1A and adrenergic alpha-1 receptors. The current study was conducted to determine which of these mechanisms underlies BMY-14802's AIM-suppressing effect. This characterization included testing the 5-HT1A agonist buspirone and multiple sigma agents. When these studies implicated a 5-HT1A mechanism, we subsequently undertook a pharmacological reversal study, evaluating whether the 5-HT1A antagonist WAY-100635 counteracted BMY-14802's AIM-suppressing effects. RESULTS: Buspirone dose-dependently suppressed AIM, supporting past findings. However, no AIM-suppressing effects were produced by drugs with effects at sigma receptors, including BD-1047, finasteride, SM-21, DTG, trans dehydroandrosterone (DHEA), carbetapentane, and opipramol. Finally, we show for the first time that the AIM-suppressing effect of BMY-14802 was dose-dependently prevented by WAY-100635 but not by the alpha-1 antagonist prazosin. CONCLUSIONS: BMY-14802 exerts its AIM-suppressing effects via a 5-HT1A agonist mechanism, similar to buspirone. Other 5-HT1A agonists have failed clinical trials, possibly due to submicromolar affinity at other receptors, including D2, which may exacerbate PD symptoms. BMY-14802 is a promising candidate for clinical trials due to its extremely low affinity for the D2 receptor and lack of extrapyramidal effects during prior clinical trials for schizophrenia. PMID- 19283367 TI - Combining silica-based adsorbents and SPME fibers in the extraction of the volatiles of beer: an exploratory study. AB - A series of silica-based materials were employed as sorbents within solid-phase microextraction vials. The aim of the study was to evaluate the effect of an additional phase on the distribution of the volatile and less volatile analytes. The adsorption of six probe molecules, namely isoamyl acetate, ethyl hexanoate (ethyl caproate), phenylethyl alcohol, ethyl octanoate (ethyl caprilate), 2 phenylethyl acetate, and ethyl decanoate, was monitored by detecting the desorbed amount on a DVD-CAR-PDMS fiber from Pilsen beer. The microextraction process involved the presence of different silica-based phases produced via different methods: xerogel produced by hydrolytic and non-hydrolytic routes, aerogel, pyrogenic, and precipitated silica. The resulting data are discussed in correlation with sorbent texture properties (specific area and pore diameter). The modification of silica with alkyl groups also affects the preconcentrated amount of the target molecules in the headspace. The presence of sorbents was shown to affect the analyte signal more than the addition of NaCl or the use of ultrasound. PMID- 19283368 TI - Principles of responsive lanthanide-based luminescent probes for cellular imaging. AB - The advent of chemical tools for cellular imaging--from organic dyes to green fluorescent proteins--has revolutionized the fields of molecular biology and biochemistry. Lanthanide-based probes are a new player in this area, as the last decade has seen the emergence of the first responsive luminescent lanthanide probes specifically intended for imaging cellular processes. The potential of these probes is still undervalued by the scientific community. Indeed, this class of probes offers several advantages over organic dyes and fluorescent proteins. Their very long luminescence lifetimes enable quantitative spatial determination of the intracellular concentration of an analyte through time-gating measurements. Their emission bands are very narrow and do not overlap, enabling the simultaneous use of multiple lanthanide probes to quantitatively detect several analytes without cross-interference. Herein we describe the principles behind the development of this class of probes. Sensors for a desired analyte can be designed by rationally manipulating the parameters that influence the luminescence of lanthanide complexes. We will discuss sensors based on varying the number of inner-sphere water molecules, the distance separating the antenna from the lanthanide ion, the energies of excited states of the antenna, and PeT switches. PMID- 19283369 TI - Sources of variability in interceptive movements. AB - In order to successfully intercept a moving target one must be at the right place at the right time. But simply being there is seldom enough. One usually needs to make contact in a certain manner, for instance to hit the target in a certain direction. How this is best achieved depends on the exact task, but to get an idea of what factors may limit performance we asked people to hit a moving virtual disk through a virtual goal, and analysed the spatial and temporal variability in the way in which they did so. We estimated that for our task the standard deviations in timing and spatial accuracy are about 20 ms and 5 mm. Additional variability arises from individual movements being planned slightly differently and being adjusted during execution. We argue that the way that our subjects moved was precisely tailored to the task demands, and that the movement accuracy is not only limited by the muscles and their activation, but also-and probably even mainly-by the resolution of visual perception. PMID- 19283370 TI - Responses of thoracic spinal interneurons to vestibular stimulation. AB - Vestibular influences on outflow from the spinal cord are largely mediated via spinal interneurons, although few studies have recorded interneuronal activity during labyrinthine stimulation. The present study determined the responses of upper thoracic interneurons of decerebrate cats to electrical stimulation of the vestibular nerve or natural stimulation of otolith organs and the anterior and posterior semicircular canals using rotations in vertical planes. A majority of thoracic interneurons (74/102) responded to vestibular nerve stimulation at median latencies of 6.5 ms (minimum of approximately 3 ms), suggesting that labyrinthine inputs were relayed to these neurons through trisynaptic and longer pathways. Thoracic interneuronal responses to vertical rotations were similar to those of graviceptors such as otolith organs, and a wide array of tilt directions preferentially activated different cells. Such responses were distinct from those of cells in the cervical and lumbar enlargements, which are mainly elicited by ear-down tilts and are synchronous with stimulus position when low rotational frequencies are delivered, but tend to be in phase with stimulus velocity when high frequencies are employed. The dynamic properties of thoracic interneuronal responses to tilts were instead similar to those of thoracic motoneurons and sympathetic preganglionic neurons. However, the preferred tilt directions of the interneurons were more heterogeneous than thoracic spinal outputs, showing that the outputs do not simply reflect an addition of local interneuronal activity. PMID- 19283371 TI - Different neural strategies for multimodal integration: comparison of two macaque monkey species. AB - The integration of neck proprioceptive and vestibular inputs underlies the generation of accurate postural and motor control. Recent studies have shown that central mechanisms underlying the integration of these sensory inputs differ across species. Notably, in rhesus monkey (Macaca mulata), an Old World monkey, neurons in the vestibular nuclei are insensitive to passive stimulation of neck proprioceptors. In contrast, in squirrel monkey, a New World monkey, stimulation produces robust modulation. This has led to the suggestion that there are differences in how sensory information is integrated during self-motion in Old versus New World monkeys. To test this hypothesis, we recorded from neurons in the vestibular nuclei of another species in the Macaca genus [i.e., M. fascicularis (cynomolgus monkey)]. Recordings were made from vestibular-only (VO) and position-vestibular-pause (PVP) neurons. The majority (53%) of neurons in both groups were sensitive to neck proprioceptive and vestibular stimulation during passive body-under-head and whole-body rotation, respectively. Furthermore, responses during passive rotations of the head-on-body were well predicted by the linear summation of vestibular and neck responses (which were typically antagonistic). During active head movement, the responses of VO and PVP neurons were further attenuated (relative to a model based on linear summation) for the duration of the active head movement or gaze shift, respectively. Taken together, our findings show that the brain's strategy for the central processing of sensory information can vary even within a single genus. We suggest that similar divergence may be observed in other areas in which multimodal integration occurs. PMID- 19283372 TI - The effects of task instructions on pro and antisaccade performance. AB - In the antisaccade task participants are required to overcome the strong tendency to saccade towards a sudden onset target, and instead make a saccade to the mirror image location. The task thus provides a powerful tool with which to study the cognitive processes underlying goal directed behaviour, and has become a widely used index of "disinhibition" in a range of clinical populations. Across two experiments we explored the role of top-down strategic influences on antisaccade performance by varying the instructions that participants received. Instructions to delay making a response resulted in a significant increase in correct antisaccade latencies and reduction in erroneous prosaccades towards the target. Instructions to make antisaccades as quickly as possible resulted in faster correct responses, whereas instructions to be as spatially accurate as possible increased correct antisaccade latencies. Neither of these manipulations resulted in a significant change in error rate. In a second experiment, participants made fewer errors in delayed pro and antisaccade tasks than in a standard antisaccade task. The implications of these results for current models of antisaccade performance, and the interpretation of antisaccade deficits in clinical populations are discussed. PMID- 19283373 TI - Effects of trkB knockout on topography and ocular segregation of uncrossed retinal projections. AB - TrkB is an important receptor for brain-derived neurotrophic factor and NT4, members of the neurotrophin family. TrkB signaling is crucial in many activity dependent and activity-independent processes of neural development. Here, we investigate the role of trkB signaling in the development of two distinct, organizational features of retinal projections--the segregation of crossed and uncrossed retinal inputs along the "lines of projection" that represent a single point in the visual field and the "retinotopic" mapping of retinofugal axons within their cerebral targets. Using anterograde tracing, we obtained quantitative measures of the distribution of retinal projections in the dorsal nucleus of the lateral geniculate body (LGd) and superior colliculus (SC) of wild type mice and mice homozygous for constitutive null mutation (knockout) of the full-length trkB receptor (trkB(FL)(-/-)). In trkB(FL)(-/-) mice, uncrossed retinal projections cluster normally but there is a topographic expansion in the distribution of these clusters across the SC. By contrast, the absence of trkB signaling has no significant effect on the segregation of crossed and uncrossed retinal projections along the lines of projection in LGd or SC. We conclude that the normal topographic organization of uncrossed retinal projections depends upon trkB signaling, whereas the segregation of crossed and uncrossed retinal projections is trkB-independent. We also found that in trkB(FL)(-/-) mice, neuronal number was reduced in the LGd and SC and in the caudate-putamen. Previous studies by ourselves and others have shown that the number of retinal ganglion cells (RGCs) is unchanged in trkB(FL)(-/-) mice. Together, these results demonstrate that there is no matching of the numbers of RGCs with neuronal numbers in the LGd or SC. PMID- 19283374 TI - Primary cultures of renal proximal tubule cells derived from individuals with primary hyperoxaluria. AB - The primary hyperoxalurias, PH1 and PH2, are inherited disorders caused by deficiencies of alanine:glyoxylate aminotransferase and glyoxylate reductase, respectively. Mutations in either of these enzymes leads to endogenous oxalate overproduction primarily in the liver, but most pathological effects are exhibited in the kidney ultimately leading to end-stage renal failure and systemic oxalosis. To provide a non-invasive means of accessing kidney cells from individuals with primary hyperoxaluria, we have derived primary cultures of renal proximal tubule cells from the urine of these patients. The cells stain positively for the epithelial markers pan-cytokeratin and zonula occludens 1 and the proximal tubule marker gamma-glutamyl transpeptidase. Mutation analysis confirmed that the cultured cells had the same genotype as the leucocytes of the patients and also expressed glyoxylate reductase at the mRNA level, illustrating their potential value as a source of renal material from these individuals. PMID- 19283375 TI - Neonatal clavicle dislocation. PMID- 19283376 TI - Bilateral sclerosing stromal tumor of the ovary in a premenarchal girl. AB - Sclerosing stromal tumor (SST) is a rare benign ovarian neoplasm classified as a type of sex cord stromal tumor that occurs predominantly in young patients. Several reports have described the US, CT and MR features of SST, but there have been no reports of a bilateral calcified SST in a child. We present a case of a bilateral SST of the ovary with calcification in a 12-year-old premenarchal girl and describe the US, CT, MR and pathological findings. PMID- 19283377 TI - Application of random amplified polymorphic DNA (RAPD) analysis coupled with microchip electrophoresis for high-resolution identification of Monascus strains. AB - Monascus fungi are commonly used for a variety of food products in Asia, and are also known to produce some biologically active compounds. Since the use of Monascus is expected to increase in food industries, strain-level identification and management of Monascus will be needed in the near future. In the present study, random amplified polymorphic DNA (RAPD) analysis coupled with microchip electrophoresis was applied for this purpose. Evaluations of the analysis stability revealed that reproducible results could be obtained, although template DNA fragmentation could influence the resulting RAPD pattern. RAPD analysis using 15 Monascus strains consisting of four species, M. ruber, M. pilosus, M. purpureus, and M. kaoliang showed that each strain generated a unique RAPD pattern, which allows strain-level identification of Monascus. In addition, the phylogenetic tree constructed from RAPD patterns reflected M. ruber-M. pilosus and M. purpureus-M. kaoliang clusters inferred from both ITS and beta-tubulin gene sequences, which indicated that the RAPD pattern could reflect their phylogenetic traits to a certain extent. On the other hand, RAPD analysis did not support the monophyletic clustering of the four Monascus species used in this study, which suggests the necessity of reexamination of species boundaries in Monascus. PMID- 19283378 TI - Novel gene clusters involved in arsenite oxidation and resistance in two arsenite oxidizers: Achromobacter sp. SY8 and Pseudomonas sp. TS44. AB - This study describes three gene clusters involved in arsenic redox transformation of two arsenite oxidizers: Achromobacter sp. SY8 and Pseudomonas sp. TS44. A 17.5 kb sequence containing the arsenite oxidase (aox) gene cluster (aoxX-aoxS-aoxR and aoxA-aoxB-aoxC-aoxD) was isolated from SY8 using a fosmid library approach. Similarly, a 14.6-kb sequence including the aox cluster (arsD-arsA-aoxA-aoxB) and the arsenic resistance (ars) gene cluster (arsC1-arsR-arsC2-ACR3-arsH-dual specificity phosphatase (DSP)-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) major facilitator superfamily (MFS)) was obtained from TS44 by inverse polymerase chain reaction (PCR). According to reverse transcription (RT) PCR experiments, SY8 aoxXSR and aoxABCD transcribed as two different transcripts in opposite directions, and TS44 aox and ars clusters transcribed as a single transcript in their respective cluster. All of these genes were found to be upregulated by the addition of arsenite [As(III)], arsenate [As(V)], and antimonite [Sb(III)], except that TS44 arsC1-arsR appeared to be expressed constitutively. The SY8 aox cluster was predicted to be regulated by a two-component signal transduction system and a potential regulatory model was proposed. The TS44 aox cluster is unusual since it contains structural genes only and arsDA in its upstream. The TS44 ars cluster includes several genes previously identified not associated with arsenic resistance or transformation. This study showed novel structures and arrangements of arsenic gene clusters associated with bacterial As(III) oxidation and As(V) reduction. PMID- 19283379 TI - Characterization of Edwardsiella tarda rpoS: effect on serum resistance, chondroitinase activity, biofilm formation, and autoinducer synthetases expression. AB - In this study, rpoS gene was identified from Edwardsiella tarda EIB202 and its functional role was analyzed by using an in-frame deletion mutant rpoS and the complemental strain rpoS (+). Compared with the wild type and rpoS (+), rpoS was impaired in terms of the ability to survive under oxidative stress and nutrient starvation, as well as the resistance to 50% serum of Scophthalmus maximus in 3 h, demonstrating essential roles of RpoS in stress adaptation. The rpoS mutant also displayed markedly increased chondroitinase activity and biofilm formation. Real-time polymerase chain reaction revealed that the expression level of quorum sensing autoinducer synthetase genes luxS and edwI was increased by 3.7- and 2.5 fold in the rpoS mutant strain. Those results suggested that rpoS might be involved in the negative or positive regulation of chondroitinase and biofilm formation, or quorum sensing networks in E. tarda, respectively. Although there were no obvious differences between the wild-type and the rpoS mutant in adherence of epithelioma papulosum cyprini (EPC) cell and in the lethality on fish model, rpoS deletion leads to the drastically reduced capacity for E. tarda to internalize in EPC cells, indicating that RpoS was, while not the main, the factor required for the virulence network of E. tarda. PMID- 19283380 TI - Case report: bilateral ischial stress fractures in an elite tennis player. AB - A case report of bilateral ischial stress fractures in an elite tennis player initially mimicking hamstring pathology is described. This is an unusual site of stress fracture. Typical sites of stress fracture are well documented; however, awareness of less common sites of stress-related bone injury can aid early diagnosis and treatment before overt fracture occurs. PMID- 19283381 TI - Human CD80/IL2 lentivirus transduced acute myeloid leukaemia cells enhance cytolytic activity in vitro in spite of an increase in regulatory CD4+ T cells in a subset of cultures. AB - Immunotherapeutic strategies are increasingly being explored as a method of enhancing anti-tumour immune responses in patients with acute myeloid leukaemia (AML). Regulatory CD4(+) T cells (Tregs) suppress effector T and natural killer (NK) cells and therefore pose a potential challenge to the efficacy of immunotherapy. AML cells transduced with a lentivirus expressing CD80 (B7.1) and IL2 (LV-CD80/IL2) are capable of stimulating T and NK cell cytotoxicity in vitro. This study examines the effect of CD80/IL2 modified AML cells on Treg number and function. We report a significant increase in the number of CD8(+) T cells (P = 0.046) CD3(-)CD56(+) NK cells (P = 0.028) and CD3(+)CD4(+)CD25(high)Foxp3(+) Tregs (P = 0.043) following stimulation for 7 days with allogeneic LV-CD80/IL2 AMLs. In contrast, autologous LV-CD80/IL2 AML cell cultures provide a weaker stimulation with a lower number of CD8(+) T cells (P = 0.011) and no change in NK cell or Treg numbers. However, an increase in cytotoxic CD8(+) T cells and NK cells are detected following both allogeneic and autologous LV-CD80/IL2 stimulation as demonstrated by an increase in IFN-gamma and CD107a expression. Despite the presence of increased numbers of Tregs with suppressive activity in a subset of cultures, increased lysis of unmodified AMLs was still achieved following allogeneic (day 0, 2.2%; day 7, 20.4%) and more importantly, autologous LV-CD80/IL2 culture in which AML patients had recently received intensive chemotherapy (day 0, 0%; day 7, 16%). Vaccination with LV-CD80/IL2 therefore provides a potential strategy to enhance anti-leukaemia immune responses without a concomitant stimulation of Treg-mediated inhibition of cytotoxic immunological responses. PMID- 19283382 TI - Incidental diagnosis of CML in a patient with anemia and IgG lamda monoclonal protein in blood. PMID- 19283383 TI - Maximum tree: a consistent estimator of the species tree. AB - We propose a model based approach to use multiple gene trees to estimate the species tree. The coalescent process requires that gene divergences occur earlier than species divergences when there is any polymorphism in the ancestral species. Under this scenario, speciation times are restricted to be smaller than the corresponding gene split times. The maximum tree (MT) is the tree with the largest possible speciation times in the space of species trees restricted by available gene trees. If all populations have the same population size, the MT is the maximum likelihood estimate of the species tree. It can be shown the MT is a consistent estimator of the species tree even when the MT is built upon the estimates of the true gene trees if the gene tree estimates are statistically consistent. The MT converges in probability to the true species tree at an exponential rate. PMID- 19283384 TI - On mathematical theory of selection: continuous time population dynamics. AB - Mathematical theory of selection is developed within the frameworks of general models of inhomogeneous populations with continuous time. Methods that allow us to study the distribution dynamics under natural selection and to construct explicit solutions of the models are developed. All statistical characteristics of interest, such as the mean values of the fitness or any trait can be computed effectively, and the results depend in a crucial way on the initial distribution. The developed theory provides an effective method for solving selection systems; it reduces the initial complex model to a special system of ordinary differential equations (the escort system). Applications of the method to the Price equations are given; the solutions of some particular inhomogeneous Malthusian, Ricker and logistic-like models used but not solved in the literature are derived in explicit form. PMID- 19283385 TI - Thoracic and abdominal MRA with gadofosveset: influence of injection rate on vessel signal and image quality. AB - The purpose of this study was to investigate the influence of different injection rates on the maximum signal intensity and the arterio-venous transit time of dynamic gadofosveset-enhanced first pass MR angiography (MRA). Twenty-one healthy male volunteers were examined with a time-resolved echo-shared MRA at 1.5 T. The volunteers were assigned into three groups using injection rates of either 1, 2 or 4 ml/s. The maximal signal enhancement and peak signal-to-noise ratio in the pulmonary trunk, the aortic arch, the abdominal aorta as well as both kidneys and lung parenchyma were analyzed. The arterio-venous transit time was determined. The time between maximal enhancement of the pulmonary trunk and the aortic arch was higher with the slow injection rate of 1 ml/s, while there were no differences in the time between maximal enhancement of the aortic arch and the abdominal aorta above or below the origin of the renal veins with all three injection rates. With the slow injection protocol of 1 ml/s a longer purely arterial phase of 10.5 s was achieved compared to 7.7 s with higher injection rates (p = 0.045). The time between maximal aortic signal intensity and maximal renal enhancement was equal for all injection protocols. PMID- 19283386 TI - Life-threatening versus non-life-threatening manual strangulation: are there appropriate criteria for MR imaging of the neck? AB - The aim of the study was to determine objective radiological signs of danger to life in survivors of manual strangulation and to establish a radiological scoring system for the differentiation between life-threatening and non-life-threatening strangulation by dividing the cross section of the neck into three zones (superficial, middle and deep zone). Forensic pathologists classified 56 survivors of strangulation into life-threatening and non-life-threatening cases by history and clinical examination alone, and two blinded radiologists evaluated the MRIs of the neck. In 15 cases, strangulation was life-threatening (27%), compared with 41 cases in which strangulation was non-life-threatening (73%). The best radiological signs on MRI to differentiate between the two groups were intramuscular haemorrhage/oedema, swelling of platysma and intracutaneous bleeding (all p = 0.02) followed by subcutaneous bleeding (p = 0.034) and haemorrhagic lymph nodes (p = 0.04), all indicating life-threatening strangulation. The radiological scoring system showed a sensitivity and specificity of approximately 70% for life-threatening strangulation, when at least two neck zones were affected. MRI is not only helpful in assessing the severity of strangulation, but is also an excellent documentation tool that is even admissible in court. PMID- 19283388 TI - [Therapy options for diseases of the lacrimal ducts]. PMID- 19283387 TI - MR imaging of cartilage and its repair in the knee--a review. AB - Chondral injuries are common lesions of the knee joint, and many patients could benefit from cartilage repair. Widespread cartilage repair techniques require sophisticated noninvasive follow-up using MRI. In addition to the precise morphological assessment of this area of cartilage repair, the cartilage's biochemical constitution can be determined using biochemical MRI techniques. The combination of the clinical outcome after cartilage repair together with the morphological and biochemical description of the cartilage repair tissue as well as the surrounding cartilage can lead to an optimal follow-up evaluation. The present article on MR imaging techniques of cartilage repair focuses on morphological description and scoring using techniques from conventional 2D through advanced isotropic 3D MRI sequences. Furthermore the ultrastructure of the repair tissue and the surrounding cartilage is evaluated in-vivo by biochemical T1-delayed gadolinium enhanced MRI of cartilage (dGEMRIC), T2 relaxation, and diffusion-weighted imaging techniques. PMID- 19283389 TI - Constipation in 44 patients implanted with an artificial bowel sphincter. AB - PURPOSE: Constipation with or without obstructed defecation (OD) is frequent in patients with artificial bowel sphincter (ABS). The aims of this study were (1) to evaluate the functional outcome of ABS based on postoperative constipation and (2) to assess pre-implantation data to predict post-implantation constipation. MATERIALS AND METHODS: Thirteen men and 31 women were followed up. Both fecal incontinence and constipation with and without OD were assessed after implantation. Pre-implantation characteristics were compared in patients with and without postoperative constipation. RESULTS: After implantation, nine patients (20.4%) had constipation without OD, 16 patients (36.4%) had OD, and 19 patients (43.2%) had neither of these. Incontinence was significantly more frequent in patients with postoperative constipation with or without OD. In these patients, an increase in preoperative constipation, anismus, and ultraslow waves was noted. CONCLUSION: Constipation with and without OD is frequent after implantation and interfere with the functional outcome of ABS. PMID- 19283390 TI - Laparoscopic colorectal resection for benign polyps not suitable for endoscopic polypectomy. AB - BACKGROUND AND AIMS: Endoscopic polypectomy still remains the cornerstone of therapy for colorectal polyps and adenomas. However, if colorectal polyps are too large or not accessible for endoscopic ablation or cannot be removed without an increased risk for perforation, operative procedures are required. In such circumstances, laparoscopic resection represents a minimally invasive alternative. MATERIALS AND METHODS: Between January 1993 and December 2004, more than 2,500 endoscopic polypectomies were performed at the Department of Surgery, University of Schleswig-Holstein, Campus Lubeck, Germany. In patients which could not be treated by endoscopic polypectomy due to size, location, and/or risk of complications, a laparoscopic colorectal resection was performed. All data were prospectively assessed in our "colorectal resection" database. RESULTS: The database analysis revealed 58 patients with endoscopically not resectable colorectal polyps who underwent a laparoscopic colorectal resection (intend to treat). In 54 patients, the operative procedure could be finished by the laparoscopic approach (study population). The conversion rate was 6.9% (four of 58). An ileocolic resection was performed in 20 patients (37.0%), and 14 patients (25.9%) underwent an anterior rectal resection. A right colectomy was necessary in 12 patients (22.2%), and six patients (11.1%) underwent a sigmoid resection. In the remaining two patients, a left colectomy and a resection of the transverse colon were performed. Intra- and postoperative complications occurred in five patients (9.3%). Perioperative mortality was not registered. The histopathological work-up revealed benign disease in all cases. CONCLUSION: Laparoscopic resection of colorectal polyps is a safe and minimally invasive technique for the management of benign colorectal tumors. Thus, the laparoscopic approach to endoscopically not resectable polyps enriches the therapeutic spectrum. PMID- 19283391 TI - Practicability and patients' subjective experiences of low-dose spinal anaesthesia using hyperbaric bupivacaine for transanal surgery. AB - PURPOSE: The safety, effectiveness and long lasting post-operative analgesia make spinal anaesthesia in saddle block technique an "ideal" method for transanal surgery. To improve patient satisfaction and offer reliable operation conditions to surgeons, this study quantifies practicability and patients' subjective experiences with this technique. METHODS: Within a 5-month period, 400 consecutive patients undergoing transanal surgery in saddle block technique were evaluated by a standardised questionnaire. RESULTS: The success rate of spinal anaesthesia was 99.5%. Side effects occurred far less frequently as mentioned in the literature. The duration of the sensory block was about twice as long as the time until first mobilisation and micturition. Despite some negative experiences during the procedure, 92% of the investigated patients would choose a saddle block again. CONCLUSIONS: Both from reasons of practicability and from patients' view, spinal anaesthesia in saddle block technique can be thoroughly recommended for transanal surgery. Patients undergoing a stapler haemorrhoidectomy should receive additional opioids. PMID- 19283392 TI - Systemic cytokine response after emergency and elective surgery for colorectal carcinoma. AB - BACKGROUND: Systemic cytokines (SC) are accepted mediators of host immune response. It is debated if long-term survival is influenced by emergency presentation of colorectal cancer, and the role of immunitary response is still unknown. The aim of this prospective study was to compare the SC response after emergency resection with that after elective resections of colorectal carcinoma. MATERIALS AND METHODS: One hundred six consecutive subjects with colorectal cancer were submitted to emergency (complete bowel obstruction; EMS, n = 50) or elective resection (ELS, n = 56) of the tumour. Sera were collected before surgery and at appropriate time points afterward and assayed for interleukin 1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and C-reactive protein (CRP). Five-year survival was analysed according to Kaplan Meier test. The Cox proportional hazard model was used for the multivariate analysis. RESULTS: Pre-operative levels of IL-1beta, IL-6 and CRP were statistically higher in the EMS group. Levels of TNF-alpha were not elevated after surgery and there was no difference between the groups. Five-year survival was significantly lower in the EMS group (p < 0.05). CONCLUSIONS: Immunitary response, as reflected by SC, was better after elective resection than after emergency resection of colorectal carcinoma and this difference may have implication in the long-term survival. PMID- 19283393 TI - Expression of EAAT-1 distinguishes choroid plexus tumors from normal and reactive choroid plexus epithelium. AB - Microscopic distinction of normal choroid plexus (CP) from choroid plexus tumors (CPT) may be difficult, especially in small samples of well-differentiated CP papillomas. So far, there are no established markers that reliably distinguish normal and neoplastic CP epithelium. Recently, a correlation between expression/function of glial glutamate transporters EAAT-1 (GLAST) and EAAT-2 (Glt-1) and tumor proliferation has been reported. Furthermore, we previously found that CPTs frequently express EAAT-1, but not EAAT-2. We now compared expression of EAAT-1, EAAT-2 and GFAP in non-neoplastic CP (n = 68) and CPT (n = 79) by immunohistochemistry. Tissue of normal CP was obtained from 50 autopsy cases (20 normal and 30 pathologic brains) and 18 neurosurgical specimens that included 17 fetal, 21 pediatric and 30 adult cases. In non-neoplastic postnatal CP (n = 51), focal expression of EAAT-1 was found in only two pediatric cases (4%). In CPT, expression of EAAT-1 was found in 64 of 79 (81%) tumor samples and was significantly age-dependent (P < 0.0001). Hence, EAAT-1 expression distinguishes neoplastic from normal CP, both in children (P = 0.0032) and in adults (P < 0.0001). Immunostaining for EAAT-2 in selected samples from cases of different ages showed that normal CP (n = 15) or CPT (n = 16) lacked EAAT-2 expression. GFAP expression was found in 3 of 32 (10%) normal CP and in 28 of 73 (38%) tumor samples. In conclusion, in contrast to neoplastic CP samples, expression of EAAT-1 is exceptionally rare in non-neoplastic CP. Thus, EAAT-1 is superior to GFAP as a helpful diagnostic tool in CP samples. PMID- 19283394 TI - Brain plasticity after ischemic stroke: an update. PMID- 19283395 TI - The potential of neural stem cells to repair stroke-induced brain damage. AB - Acute injuries to CNS such as stroke induce neural progenitor proliferation in adult brain which might be an endogenous attempt to self-repair. This process is known to be altered by several exogenous and endogenous modulators including growth factors that could help to reinforce the post-stroke neurogenesis. Increasing the neurogenesis may be a future therapeutic option to decrease the cognitive and behavioral deficits following stroke. In addition, transplantation of various types of stem cells into the injured brain is currently thought to be an exciting option to replace the neurons lost in the post-ischemic brain. These include immortalized stem cell lines, neural progenitors prepared from embryonic and adult animals and mesenchymal stem cells. Using exogenous stem cells in addition to modulating endogenous neurogenesis, we may be able to repair the injured brain after a devastating stroke. This article reviewed the current literature of these two issues. PMID- 19283396 TI - TDP-43 pathology in familial British dementia. AB - Trans-activation-responsive DNA-binding protein 43 (TDP-43) is a component of pathological inclusions in amyotrophic lateral sclerosis and several forms of sporadic and familial frontotemporal lobar degeneration. This has suggested defining a new class of diseases known as TDP-43 proteinopathies. However, it has been reported more recently that TDP-43 positive inclusions occur in other neurodegenerative disorders such as Alzheimer's disease, Dementia with Lewy Bodies and Parkinsonism dementia complex of Guam. Here we report the occurrence of TDP-43 inclusions in one other neurodegenerative disorder: familial British dementia. Using a variety of antibodies against phosphorylated and non phosphorylated TDP-43 epitopes, we found intense accumulation occurred in the form of dystrophic neurites, neuronal cytoplasmic inclusions and was also occasionally associated with neurofibrillary tangles. Double immunostaining revealed that TDP-43 and tau aggregates were rarely directly colocalized, but co existed in the same neurons as separate inclusions. Double staining with ubiquitin showed a direct colocalization with TDP-43. The phosphorylation dependent TDP-43 antibodies proved superior to phosphorylation-independent antibodies in revealing pathological inclusions since the former did not stain non-phosphorylated TDP-43 in normal nuclei. Our results support the concept that TDP-43 pathology is not narrowly restricted, but is involved in the etiology of many neurodegenerative disorders. PMID- 19283397 TI - Onset of labour: women's experiences and midwives' assessments in relation to first stage duration. AB - AIM: The study aimed to assess the time of labour onset and its symptoms as perceived by women in labour and midwives, and the relationship between these and first stage duration. METHODS: A longitudinal cohort study of women with a singleton pregnancy in cephalic presentation was performed in 41 maternity units. The sample comprised 1,170 women, 611 nulliparae (np) and 559 multiparae (mp), who answered two standardized questions on the onset of labour and selected the applicable symptoms of labour onset from a list of eight. A multivariate Cox regression model was computed covering further perinatal factors. RESULTS: The median durations of the first stage of labour as assessed by the women themselves were 11 (np) and 6.5 h (mp), and as assessed by the midwives 7 (np) and 4 h (mp). Median time intervals between the start of labour onset symptoms as perceived by the women concerned and the midwives' diagnoses varied greatly: the shortest related to watery fluid loss (np = 1.5 h, mp = 0.0 h), the longest to alterations in sleep patterns (np = 11.5 h, mp = 4.5 h). Irregular pain, watery fluid loss and the time between self-diagnosed and professionally diagnosed onset of labour were just as closely associated with the duration of the first stage of labour as perinatal factors. Significant perinatal factors were induction with oxytocin, herbal remedies and PROM. CONCLUSIONS: The perceptions of women in labour are as important as perinatal factors in determining the duration of the first stage of labour and should be taken into account in intrapartum care. PMID- 19283398 TI - Obstetric morbidity and the diagnostic dilemma in pregnancy in rudimentary horn: retrospective analysis. AB - BACKGROUND: Pregnancy in rudimentary horn of uterus, a form of ectopic gestation, is associated with significant rates of morbidity and mortality. Despite the recent advances in the ultrasonography, diagnosis of cornual pregnancy still remains elusive; with confirmatory diagnosis usually made during laparotomy. The aim of the present study is to analyze the obstetric implications and the diagnostic dilemma of rudimentary horn pregnancy. MATERIALS AND METHODS: Records of women diagnosed with ectopic pregnancy in the rudimentary horn, during the years 2004 to 2008, managed in a referral hospital in northern India; were reviewed for their diagnostic difficulties and the associated morbidity. RESULTS: During the four year study period, rudimentary horn pregnancies accounted for 12 pregnancies. Non communicating horn accounted for 75% of the cases. The mean age of women at presentation was 26 +/- 5.11 years and the period of gestation at diagnosis varied between 10 and 34 weeks. Preruputure diagnosis was possible only in two cases and sensitivity of ultrasonographic diagnosis was 33.3%. Laprotomy with excision of rudimentary horn and salpingectomy was done in all cases. Multiple blood transfusions were required in 83.3% of women. CONCLUSION: Management of pregnancy in a rudimentary uterine horn continues to be a challenge to this day. Maintaining a higher degree of alertness, especially in high risk groups by emergency staff is required to prevent the morbidity, as they may present with acute uterine rupture in pregnancy. PMID- 19283399 TI - Pharyngocutaneous fistulae following total laryngectomy comparison between manual and mechanical sutures. AB - The extension of the surgery and closure type of the pharynx can be the determinants in the pharyngocutaneous fistula development. The objective of the study is to evaluate the incidence of pharingocutaneous salivary fistulae after total laryngectomies comparing manual and mechanical sutures. The study is designed as non-randomized, prospective clinical study. Sixty patients with squamous cell carcinoma were submitted to total laryngectomies. In 30 cases, the linear stapler (75 mm) closure (surgical technique described in details) and in other 30 cases manual suture was used. The cases of mechanical suture were prospective and consecutive and the cases of manual suture were a review series of patients who underwent a manual suture of pharynx, in the same period of time. The statistical analysis between the two groups concluded that both were comparable. Fistulae incidence was 6.7% (2/30) in the group with the mechanical suture and 36.7% (11/30) in the group with manual suture closure, presenting a significant difference (p = 0.0047). The total laryngectomy with mechanical closure is an easy and fast learning technique, allowing watertight closure of the pharynx with a low risk of contamination of the surgical field. It is an assured method, even in previously irradiated patients, since we respect the limits of its indication regarding the extension of primary tumor that must be confirmed by previous suspension laryngectomy performed in the operating room. PMID- 19283400 TI - Rehabilitation and outcome of severe profound deafness in a group of 16 infants affected by congenital cytomegalovirus infection. AB - The aim of the study was to characterize the audiological consequences of congenital cytomegalovirus infection (CMV) and to evaluate the outcome of rehabilitation with hearing aids and/or cochlear implant (CI), associated with an adequate speech-language therapy. A retrospective review of data was made from a total of 16 infants, affected by severe to profound hearing loss from congenital CMV infection, referred to a tertiary audiological center for rehabilitation. Audiological evaluation was performed using behavioral audiometry, auditory brainstem responses (ABR) and/or electrocochleography (ECochG). Of the 16 children (median age at diagnosis of hearing loss: 21.33 +/- 0.7 months) with CMV hearing loss, 14 were affected by profound bilateral hearing loss and received a CI, while 2 were affected by bilateral severe hearing loss and received hearing aids. Cochlear implants can provide useful speech comprehension to patients with CMV-related deafness, even if language development is lower when compared to a group of Connexin (Cx) 26+ cochlear-implanted children (eight subjects), matched for age. Congenital CMV infection still represents a serious clinical condition, as well as an important cause of hearing loss in children. More studies have claimed to identify the pathophysiological mechanisms of damage and thus to ensure a better therapeutic approach. Nonetheless, in cases of CMV-deafened babies, the overall outcome of cochlear implantation is good. PMID- 19283401 TI - MMP-14 can serve as a prognostic marker in patients with supraglottic cancer. AB - The purpose of this study was to examine the expression of MMP-14, 15 and 16 (MT1, MT2 and MT3-MMP) in supraglottic carcinoma and to explore their clinical significance. Expression of MMP-14, 15 and 16 was examined using RT-PCR and immunohistochemistry methods in samples from 85 cases of supraglottic carcinoma. Results showed that both mRNA and protein expression of MMP-14, 15 and 16 were increased in supraglottic carcinoma tissues compared to expression in control adjacent non-neoplastic tissues (P < 0.05). Expression of MMP-14, but not MMP-15 and MMP-16, was significantly increased in the T3 and neck nodal metastasis groups compared with the T1-2 group and the group without nodal metastasis at both mRNA and protein levels (P < 0.05). Expression of MMP-14 mRNA and protein was also higher in tumors of patients with stage III-IV disease compared to patients with clinical stage I-II tumors (P < 0.05). Kaplan-Meier survival curves showed that the groups with high MMP-14 protein expression had a poorer prognosis than patients in the groups with weak or negative expression of MMP-14 protein (P < 0.05). In conclusion, MMP-14 may play an important role in the progression of supraglottic carcinoma and may be a novel prognostic factor for patients with supraglottic carcinoma. PMID- 19283402 TI - Drainage of the tracheal blind pouch created by laryngotracheal separation. AB - Laryngotracheal separation is a simple and reliable operation for the treatment of patients with repetitive and intractable aspiration; however, it is apprehended that pooling in the tracheal blind pouch may cause postoperative complications. In the present study, we examined drainage of the blind pouch created by laryngotracheal separation. Fourteen patients aged 3-63 years with repetitive aspiration pneumonia underwent laryngotracheal separation by the modified Lindeman procedure. A barium swallow was performed 10-30 days after surgery. X-rays of the lateral view of the neck were taken at 6 and 24 h after the swallow, and then every 24 h until the contrast medium cleared. The contrast medium in the blind pouch cleared within 24 h in nine patients. In the remaining five, the clearance time was < or =48 and < or =72 h in two patients each, and 96 h in one patient. The clearance time in patients aged under 20 years was < or =24 h, while middle-aged to elderly patients showed prolonged clearance time. No late complications of the blind pouch, such as infections, were observed. The potential risk of complications caused by pooling in the tracheal blind pouch in laryngotracheal separation is prevented presumably due to the slow but continuous turnover of pooling material. This result supports the validity and usefulness of laryngotracheal separation for the treatment of intractable aspiration. PMID- 19283403 TI - The growth rate and size of the mastoid air cell system and mastoid bone: a review and reference. AB - This review suggests a reference to the postnatal growth of mastoid air cells and bone. Information was retrieved from studies having large consecutive age groups, in order to reveal a development pattern. Data regarding origin, gender, and antibiotic treatment was investigated as well. Most measurements were obtained by planimetry. Assessment of the various data sources suggested the antrum to be well developed at birth (1-1.5 cm2), the mastoid cells to be about 3.5-4 cm2 at 1 year, followed by a linear growth till the age of 6 (1-1.2 cm2/year), having a slower increment up to adult size at puberty (approximately 12 cm2). The mastoid bone expansion is about 0.6-0.9 cm/year in length and width and 0.4 cm/year in depth in the first year, followed by half that rate until the age of 6-7. At puberty there was a slower sprout reaching adult size. Different ethnic groups share similar mastoid aeration and bone growth patterns. There were no differences between mastoid aeration measured at the pre-antibiotic era and after its widespread use. In conclusion, there are three distinguishable phases of mastoid pneumatization from birth till reaching final size. Bone and air cell compartments share a similar growth pattern; bone expansion lags behind aeration. Antibiotic treatment for otitis may have no impact upon mastoid aeration. PMID- 19283404 TI - Morphological evidence of biofilm formation in Greenlanders with chronic suppurative otitis media. AB - Biofilm may explain the recurrences and recalcitrant episodes of otorrhea in chronic suppurative otitis media (CSOM). This study investigates bacterial biofilm in Greenlanders with CSOM and chronic otitis media with effusion (COME). The study is partly blinded, prospective and retrospective. Six children with CSOM, four children with COME and ten adults with CSOM were included in this study. Cultures were obtained and examined by standard methods. Otorrhea or glue was collected from the children and smears were prepared. Middle ear mucosa biopsies were obtained from the adults. Smears and biopsies were analyzed with microscopy and peptide nucleic acid fluorescence in situ hybridization (PNA FISH). Biofilm was confirmed in 83% of CSOM smears but in none of the COME smears. Mucosal biofilm was confirmed in 80% of the biopsies from adults with CSOM. This study provides direct morphological evidence of biofilm in samples from human CSOM. This may help to explain the microbiological mechanisms of the disease and alter the treatment strategy in the future. PMID- 19283405 TI - How long should the sudden hearing loss patients be followed after early steroid combination therapy? AB - To evaluate the beginning time and the completion time of hearing improvement in patients with sudden hearing loss who were treated with combination therapy including oral steroid. From September 2006 to December 2007, 102 idiopathic sudden hearing loss patients who showed any 'recovery' in hearing according to the Siegel's criteria after treatment were analyzed. Pure tone audiometries were performed on the pretreatment day and on the third, seventh, fourteenth post treatment day, and on the first, second, third, sixth post-treatment month. The time of initial hearing improvement and the completion time of hearing improvement were analyzed. Of 102 patients who showed any improvement, cumulatively, 93.1% showed beginning of hearing improvement within 14 days after treatment. Complete recovery or an end of change was achieved in cumulatively 80.4% of the patients within 1 month after treatment and in 92.2% of the patients within 2 months after treatment. Prognosis can be predicted approximately 2 weeks after start of treatment because time of commencement shows plateau after 2 weeks in improved cases. Hearing should be followed-up for at least 2 months after treatment in patients who show incomplete or delayed hearing improvement. PMID- 19283406 TI - Thyroarytenoid muscle invasion in T1 glottic carcinoma. AB - We report the prevalence of thyroarytenoid (TA) muscle invasion in 109 consecutive patients with T1 glottic carcinoma submitted to endoscopic laser cordectomy between February 1997 and January 2006, in order to evaluate if routine resection of the TA is necessary. A total of 109 patients staged as T1 underwent endoscopic treatment with carbon dioxide laser (CO(2)), 36 cases were treated with type I or II cordectomies, and 73 patients underwent type III, IV or V cordectomies, with resection of all or part of the TA. Over a total of 109 patients, 6 (5.5%) cases staged as pT1 showed TA invasion. The endoscopic treatment of T1 glottic cancer should be as conservative as possible in terms of TA resection, since muscle invasion is rare. In many cases, type III and IV cordectomies can be regarded as excessive treatment. Muscle invasion found histologically after type II cordectomy can be managed by further excision. PMID- 19283407 TI - Psychomotor developmental delay and epilepsy in an offspring of father-daughter incest: quantification of the causality probability. AB - A 20-year-old offspring of father-daughter incest, who has been suffering from serious psychomotoric health problems since early childhood, is seeking financial compensation under the German federal act of victim indemnification. For her appeal to be valid, the probability X that the incest was causal for her disorder must exceed 50%. Based upon the available medical records, we show that this is indeed the case and that X is even likely to exceed 65%, thereby rendering the victim's claim scientifically and legally justified. PMID- 19283408 TI - Cloning and characterization of a squalene synthase gene from a petroleum plant, Euphorbia tirucalli L. AB - Euphorbia tirucalli L., which is also known as a petroleum plant, produces a large amount of phytosterols and triterpenes. During their biosynthesis, squalene synthase converts two molecules of the hydrophilic substrate farnesyl diphosphate into a hydrophobic product, squalene. An E. tirucalli cDNA clone of a putative squalene synthase gene (EtSS) was isolated by RT-PCR followed by 5'- and 3'-RACE. The restriction fragment polymorphisms revealed by Southern blot analysis suggest that EtSS is a single copy gene. The glycine at the 287th residue from the N terminal end of domain C has replaced alanine, which is conserved among all the other SS sequences deposited in the Genbank database. The N-terminal 380 residues of the hydrophilic sequence was expressed as a peptide-tagged protein in E. coli, and the resultant bacterial crude extract was incubated with farnesyl diphosphate and NADPH. GC-MS analysis showed that squalene was detected in the in vitro reaction mixture. E. tirucalli transgenic callus lines, in which EtSS was overexpressed, accumulated increased amounts of phytosterols as compared with that of wild type callus. RT-PCR analysis of wild type E. tirucalli plants revealed that the EtSS transcript accumulated in almost equal amounts in the stems and the leaves with a stalk, while a lower amount was detected in the roots. In situ hybridization analysis revealed that prominent antisense-probe signal was detected in the cambia within bundle sheathes. These results indicate that EtSS functions prominently in cambia, which are located adjacent to conductive tubes, and that this gene plays important roles in phytosterol accumulation in petroleum plants. PMID- 19283409 TI - Perspectives on the Trypanosoma cruzi-host cell receptor interactions. AB - Chagas disease is caused by the parasite Trypanosoma cruzi. The critical initial event is the interaction of the trypomastigote form of the parasite with host receptors. This review highlights recent observations concerning these interactions. Some of the key receptors considered are those for thromboxane, bradykinin, and for the nerve growth factor TrKA. Other important receptors such as galectin-3, thrombospondin, and laminin are also discussed. Investigation into the molecular biology and cell biology of host receptors for T. cruzi may provide novel therapeutic targets. PMID- 19283410 TI - High-resolution mapping of the S-locus in Turnera leads to the discovery of three genes tightly associated with the S-alleles. AB - While the breeding system known as distyly has been used as a model system in genetics, and evolutionary biology for over a century, the genes determining this system remain unknown. To positionally clone genes determining distyly, a high resolution map of the S-locus region of Turnera has been constructed using segregation data from 2,013 backcross progeny. We discovered three putative genes tightly linked with the S-locus. An N-acetyltransferase (TkNACE) flanks the S locus at 0.35 cM while a sulfotransferase (TkST1) and a non-LTR retroelement (TsRETRO) show complete linkage to the S-locus. An assay of population samples of six species revealed that TsRETRO, initially discovered in diploid Turnera subulata, is also associated with the S-allele in tetraploid T. subulata and diploid Turnera scabra. The sulfotransferase gene shows some level of differential expression in long versus short styles, indicating it might be involved in some aspect of distyly. The complete linkage of TkST1 and TsRETRO to the S-locus suggests that both genes may reside within, or in the immediate vicinity of the S-locus. Chromosome walking has been initiated using one of the genes discovered in the present study to identify the genes determining distyly. PMID- 19283411 TI - Determinants and implications of cancer patients' psychosocial needs. AB - GOALS OF WORK: Cancer patients often experience distress. However, the majority of newly diagnosed patients gradually adapt to the crisis. When symptoms of distress and anxiety persist over months, patients require psychosocial support. The aim of the present study was to determine the proportion of cancer patients who indicate the need for psychosocial support and to identify sociodemographic, psychological and illness-related factors predicting the need for psychosocial support in a German sample. MATERIALS AND METHODS: The cross-sectional retrospective study was administered to 710 cancer patients who had been inpatients at the University Hospital of Cologne. The response rate was 49.5%. Patients suffering from bronchial, oesophageal, colorectal, breast, prostate and skin cancer participated in the study. The severity of depressive symptoms was assessed using the German version of the Major Depression Inventory. The level of anxiety was assessed with the state subscale of the German version of the State Trait Anxiety Inventory. To measure the functional aspects of health-related quality of life, the scales "physical functioning", "role functioning", "emotional functioning", "cognitive functioning" and "role functioning" of the European Organisation for Research and Treatment of Cancer QLQ-C30 Questionnaire (EORTC QLQ-C30) were used. MAIN RESULTS: Of the cancer patients, 18.9% indicate an unmet need for psychosocial support and 9.5% are actually using psychosocial services. In a multiple logistic regression, significant indicators of the need for psychosocial support are gender [p = 0.014; standardised effect coefficient (sc) = 1.615] and emotional functioning (p < 0.001; sc = 1.533). The estimated model has a specificity of 92.2% and a sensitivity of 54.0%. CONCLUSION: Almost a third of the cancer patient population indicates an unmet need for psychosocial support or is actually using psychosocial services. Emotional functioning is a central predictor of the requirement for psychosocial support. Women are emotionally more affected than men and need more psychosocial support. The prognostic validity of the severity of depression and anxiety is limited. PMID- 19283412 TI - Depression in lung cancer patients: is the HADS an effective screening tool? AB - INTRODUCTION: The present study aimed at comparing the efficacy in recognizing depression, in 53 patients newly diagnosed with lung cancer, of the Hospital Anxiety and Depression Scale (HADS), a self-report screening questionnaire, and the Montgomery-Asberg Depression Rating Scale (MADRS), a semi-structured clinician-rated interview. Specifically, we aimed at addressing the question of which is the best HADS cutoff for the detection of patients to be further investigated through a clinical semi-structured interview (the MADRS). RESULTS: The MADRS identified 92% of the patients as depressed; the HADS, 70% and 87%, using a cutoff of 11 and 8, respectively. The results suggest substantial agreement between the HADS and the MADRS when a cutoff of 8 is used (McNemar: p = 0.51; Cohen K = 0.69), while a HADS cutoff of 11 resulted in a significantly lower concordance with the MADRS (McNemar: p = 0.002; Cohen K = 0.49). PMID- 19283413 TI - Effect of fibrin glue on the prevention of persistent cerebral spinal fluid leakage after incidental durotomy during lumbar spinal surgery. AB - Approximately one million spinal surgeries are performed in the United States each year. The risk of an incidental durotomy (ID) and resultant persistent cerebrospinal fluid (CSF) leakage is a significant concern for surgeons, as this complication has been associated with increased length of hospitalization, worse neurological outcome, and the development of CSF fistulae. Augmentation of standard dural suture repair with the application of fibrin glue has been suggested to reduce the frequency of these complications. This study examined unintended durotomies during lumbar spine surgery in a large surgical patient cohort and the impact of fibrin glue usage as part of the ID repair on the incidence of persistent CSF leakage. A retrospective analysis of 4,835 surgical procedures of the lumbar spine from a single institution over a 10-year period was performed to determine the rate of ID. The 90-day clinical course of these patients was evaluated. Clinical examination, B-2 transferrin assay, and radiographic imaging were utilized to determine the number of persistent CSF leaks after repair with or without fibrin glue. Five hundred forty-seven patients (11.3%) experienced a durotomy during surgery. Of this cohort, fibrin glue was used in the dural repair in 278 patients (50.8%). Logistic models evaluating age, sex, redo surgery, and the use of fibrin glue revealed that prior lumbar spinal surgery was the only univariate predictor of persistent CSF leak, conferring a 2.8-fold increase in risk. A persistent CSF leak, defined as continued drainage of CSF from the operative incision within 90 days of the surgery that required an intervention greater than simple bed rest or over-sewing of the wound, was noted in a total of 64 patients (11.7%). This persistent CSF leak rate was significantly higher (P < 0.001) in patients with prior lumbar surgery (21%) versus those undergoing their first spine surgery (9%). There was no statistical difference in persistent CSF leak between those cases in which fibrin glue was used at the time of surgery and those in which fibrin glue was not used. There were no complications associated with the use of fibrin glue. A history of prior surgery significantly increases the incidence of durotomy during elective lumbar spine surgery. In patients who experienced a durotomy during lumbar spine surgery, the use of fibrin glue for dural repair did not significantly decrease the incidence of a persistent CSF leak. PMID- 19283414 TI - Demographics of acute admissions to a National Spinal Injuries Unit. AB - This prospective demographic study was undertaken to review the epidemiology and demographics of all acute admissions to the National Spinal Injuries Unit in Ireland for the 5 years to 2003. The study was conducted at the National Spinal Injuries Unit, Mater Miscericordiae University Hospital, Dublin, Ireland. Records of all patients admitted to our unit from 1999 to 2003 were compiled from a prospective computerized spinal database. In this 5-year period, 942 patients were acutely hospitalized at the National Spinal Injuries Unit. There were 686 (73%) males and 256 (27%) females, with an average age of 32 years (range 16-84 years). The leading cause of admission with a spinal injury was road traffic accidents (42%), followed by falls (35%), sport (11%), neoplasia (7.5%) and miscellaneous (4.5%). The cervical spine was most commonly affected (51%), followed by lumbar (28%) and thoracic (21%). On admission 38% of patients were ASIA D or worse, of which one-third were AISA A. Understanding of the demographics of spinal column injuries in unique populations can help us to develop preventative and treatment strategies at both national and international levels. PMID- 19283415 TI - Ashkenazi Parkinson's disease patients with the LRRK2 G2019S mutation share a common founder dating from the second to fifth centuries. AB - The LRRK2 G2019S mutation is a major genetic determinant of Parkinson's disease (PD) across the world that occurs at an elevated frequency in Ashkenazi Jews. We determined the LRRK2 haplotypes in 77 G2019S carriers, mostly Ashkenazi Jews, and in 50 noncarrier Ashkenazi PD patients, using 16 genetic markers. A single haplotype was detected in all mutation carriers, indicating that these individuals share a common founder. Using a maximum-likelihood method, we estimate that Ashkenazi Jews with G2019S share a common ancestor who lived approximately 1,830 (95% CI 1,560-2,160) years ago, around the second century, after the second Jewish Diaspora. PMID- 19283416 TI - Socially dependent auditory laterality in domestic horses (Equus caballus). AB - Laterality is now known to be an ubiquitous phenomenon among the vertebrates. Particularly, laterality of auditory processing has been demonstrated in a variety of species, especially songbirds and primates. Such a hemispheric specialization has been shown to depend on factors such as sound structure, species specificity and types of stimuli. Much less is known on the possible influence of social familiarity although a few studies suggest such an influence. Here we tested the influence of the degree of familiarity on the laterality of the auditory response in the domestic horse. This species is known for its social system and shows visible reactions to sounds, with one or two ears moving towards a sound source. By comparing such responses to the playback of different conspecific whinnies (group member, neighbor and stranger), we could demonstrate a clear left hemisphere (LH) preference for familiar neighbor calls while no preference was found for group member and stranger calls. Yet, we found an opposite pattern of ear side preference for neighbor versus stranger calls. These results are, to our knowledge, the first to demonstrate auditory laterality in an ungulate species. They open further lines of thought on the influence of the social "value" of calls and the listener's arousal on auditory processing and laterality. PMID- 19283417 TI - Perceived job security and sickness absence: a study on moral hazard. AB - A moral hazard problem was investigated by analysing the individual behaviour of female and male employees with regard to utilisation of sickness insurance in connection with perceived job security. It was hypothesised that employees with a higher perceived job security take more frequent sickness absence. Perceived higher job security is indicated by three variables, namely a permanent job contract, no unemployment history, and native ethnicity. The effect of perceived job security is expected to be stronger on short-term than on long-term sickness absence, since a medical certificate is required for the latter. Public health survey data from Stockholm County, Sweden, covering the year 2002 was used. Using logistic regression analyses separately for short- and long-term sickness absence and for females and males, we found that short-term sickness absence is more strongly influenced by perceived job security than long-term sickness absence. We observe indications of moral hazard in both female and male employees. However, the three indicators of perceived job security have a different influence on females and males. PMID- 19283418 TI - Different roads to discovery; Prontosil (hence sulfa drugs) and penicillin (hence beta-lactams). AB - The important chemotherapeutic agents, Prontosil and pentenylpenicillin (penicillin F), were investigated initially by two men, Domagk and Fleming, who had been influenced by the horrendous wound infections of World War I. The very different pathways leading to their development and to that of the successor antibacterials (sulfa drugs, further penicillins, semi-synthetic penicillins), including the role played by patents, are discussed. PMID- 19283419 TI - Media optimization for biosurfactant production by Rhodococcus erythropolis MTCC 2794: artificial intelligence versus a statistical approach. AB - This paper entails a comprehensive study on production of a biosurfactant from Rhodococcus erythropolis MTCC 2794. Two optimization techniques--(1) artificial neural network (ANN) coupled with genetic algorithm (GA) and (2) response surface methodology (RSM)--were used for media optimization in order to enhance the biosurfactant yield by Rhodococcus erythropolis MTCC 2794. ANN and RSM models were developed, incorporating the quantity of four medium components (sucrose, yeast extract, meat peptone, and toluene) as independent input variables and biosurfactant yield [calculated in terms of percent emulsification index (% EI(24))] as output variable. ANN-GA and RSM were compared for their predictive and generalization ability using a separate data set of 16 experiments, for which the average quadratic errors were approximately 3 and approximately 6%, respectively. ANN-GA was found to be more accurate and consistent in predicting optimized conditions and maximum yield than RSM. For the ANN-GA model, the values of correlation coefficient and average quadratic error were approximately 0.99 and approximately 3%, respectively. It was also shown that ANN-based models could be used accurately for sensitivity analysis. ANN-GA-optimized media gave about a 3.5-fold enhancement in biosurfactant yield. PMID- 19283420 TI - Effects of triclosan on zebrafish early-life stages and adults. AB - BACKGROUND, AIM AND SCOPE: The biocide triclosan (TCS) is commonly used in personal care, acrylic, plastic, and textiles products. TCS has been detected in surface water in several countries, and its ecological impact is largely unknown. In this work, the toxicity of TCS in zebrafish (Danio rerio), embryos and adults was studied. Several lethal and sub-lethal endpoints were analysed in organisms exposed to TCS such as mortality, embryo development and behaviour, hatching, micronuclei and biochemical markers (cholinesterase (ChE), glutathione S transferase (GST) and lactate dehydrogenase (LDH)). MATERIALS AND METHODS: Embryo/larvae assay followed the OECD guideline on Fish Embryo Toxicity Test. Embryos were exposed at nominal concentrations of 0.1, 0.3, 0.5, 0.7 and 0.9 mg/l of TCS for 6 days and were inspected daily with the help of a stereomicroscopy for mortality, developmental parameters (otolith formation, eye and body pigmentation, somite formation, heart beat, tail circulation, detachment of the tail-bud from the yolk sac) and hatching. A similar test was run to obtain larvae for ChE, GST and LDH analysis. The adult test followed the OECD Guideline TG 203 in semi-static conditions. Adult zebrafish of similar length and age were exposed to nominal concentrations of 0.1, 0.2, 0.3, 0.4 and 0.5 mg/l of TCS for 96 h and were inspected daily for mortality and behaviour alterations. A second test was run to obtain organs for biomarkers analysis: Heads, muscles and gills were isolated and snap-frozen in eppendorfs and used for ChE, LDH and GST determinations, respectively. Adult zebrafish testing also comprised a third test for micronucleus analysis in which the nominal concentrations of 0, 0.175 and 0.350 mg/l were used. Peripheral blood was obtained by cardiac puncture and used for the analysis. RESULTS: TCS showed acute toxicity for embryo/larvae (96 h LC(50) = 0.42 mg/l) and delayed hatching. Moreover, embryo toxicity was evident: Delay on the otolith formation and eye and body pigmentation were found, and malformations were also evident, including spine malformations, pericardial oedema and undersize. Biomarkers levels were affected: ChE and LDH activity were increased in larvae exposed to 0.25 mg/l, and GST activity was increased in larvae exposed to 0.25 and 0.35 mg/l. TCS also demonstrated acute toxicity to adult zebrafish (96 h LC(50) = 0.34 mg/l). However, TCS did not change biomarkers levels and did not elicit a micronucleus in adults. DISCUSSIONS: Despite the fact that similar 96 h LC(50) values have been found for D. rerio embryos and adults (0.42 and 0.34 mg/l, respectively), the embryo assay was much more informative, showing important effects at several levels, including teratogenic response, hatching delay and alteration of biomarker levels. TCS does not seem to be genotoxic for adult fish or to interfere with biomarkers levels at the concentrations tested. CONCLUSIONS: TCS has deleterious effects on zebrafish adults and during early stages, (including embryotoxicity, hatching delay and alterations of biomarkers levels). The range of endpoints used on the embryo test allows an integrated analysis that contributes to a better understanding of the toxicity and mode of action of TCS. RECOMMENDATIONS AND PERSPECTIVES: Future works should focus on a deeper investigation of TCS modes of action on zebrafish early-life stages. As embryo testing was revealed to be so informative, a refinement of the test could be made, including other endpoints such as different biochemical markers as well as DNA microarrays to assess a gene expression level for the effect of exposure to TCS. In the perspective of risk assessment, these endpoints should be explored in order to assess their usefulness as early warning signs and links should be sought between these short-term tests and effects of long-term exposures as it is observed in more realistic scenarios. PMID- 19283421 TI - Microbubbles traversing the blood-brain barrier for imaging and therapy. AB - In the last several years great progress has been made in the field of ultrasound perfusion imaging of the brain. Different approaches have been assessed and shown to be capable of early detection of cerebral perfusion deficits. Real-time low mechanical index imaging simplifies the acquisition of perfusion parameters and alleviates many of the previous imaging problems related to shadowing, uniplanar analysis, and temporal resolution. With the advent of this new, highly sensitive contrast-specific imaging technique new possibilities of real-time visualization of brain infarctions and cerebral hemorrhages have emerged. Microbubbles that traverse the blood-brain barrier (BBB) can also elicit bioeffects that may be used to open the BBB for targeted delivery of macromolecular agents to the brain. Possible ways in which substances cross the BBB after application of this novel approach include transcytosis, passage through endothelial cell cytoplasmic openings, opening of tight junctions, and free passage through injured endothelium. Although relatively little tissue damage occurs at low acoustic intensities capable of opening the BBB, no investigation has demonstrated a total lack of BBB injury when using ultrasound and microbubbles. Further studies are necessary to address the effects of ultrasound and microbubbles upon the various transport mechanisms of the BBB. Moreover, investigations aimed at elucidating how ultrasound and microbubbles interact at the molecular level of the BBB are necessary. Results of such studies will increase our understanding of the mechanisms of BBB opening and also allow a better appraisal of the safety of this technique for future clinical applications. PMID- 19283422 TI - Effects of different technical coordinate system definitions on the three dimensional representation of the glenohumeral joint centre. AB - This study aimed to find the most appropriate marker location, or combination thereof, for the centre of the humeral head (Wang et al. in J Biomech 31: 899 908, 1998) location representation during humeral motion. Ten male participants underwent three MRI scans in three different humeral postures. Seven technical coordinate systems (TCS) were defined from various combinations of an acromion, distal upper arm and proximal upper arm clusters of markers in a custom Matlab program. The CHH location was transformed between postures and then compared with the original MRI CHH location. The results demonstrated that following the performance of two near 180 degrees humeral elevations, a combined acromion TCS and proximal upper arm TCS produced an average error of 23 +/- 9 mm, and 18 +/- 4 mm, which was significantly smaller (p < 0.01) than any other TCS. A combination of acromion and proximal upper arm TCSs should therefore be used to reference the CHH location when analysing movements incorporating large ranges of shoulder motion. PMID- 19283423 TI - The effect of trans-10, cis-12 conjugated linoleic acid on gene expression profiles related to lipid metabolism in human intestinal-like Caco-2 cells. AB - We conducted an in-depth investigation of the effects of conjugated linoleic acid (CLA) on the expression of key metabolic genes and genes of known importance in intestinal lipid metabolism using the Caco-2 cell model. Cells were treated with 80 mumol/L of linoleic acid (control), trans-10, cis-12 CLA or cis-9, trans-11 CLA. RNA was isolated from the cells, labelled and hybridized to the Affymetrix U133 2.0 Plus arrays (n = 3). Data and functional analysis were preformed using Bioconductor. Gene ontology analysis (GO) revealed a significant enrichment (P < 0.0001) for the GO term lipid metabolism with genes up-regulated by trans-10, cis 12 CLA. Trans-10, cis-12 CLA, but not cis-9, trans-11 CLA, altered the expression of a number of genes involved in lipid transport, fatty acid metabolism, lipolysis, beta-oxidation, steroid metabolism, cholesterol biosynthesis, membrane lipid metabolism, gluconeogenesis and the citrate cycle. These observations warrant further investigation to understand their potential role in the metabolic syndrome. PMID- 19283426 TI - A delay in pubertal onset affects the covariation of body weight, estradiol, and bone size. AB - The skeletal system functions as a locomotive organ and a mineral reservoir and combinations of genetic and environmental factors affect the skeletal system. Although delayed puberty is associated with compromised bone mass, suppression of estrogen should be beneficial to cortical strength. The purpose was to employ path analysis to study bone strength and delayed puberty. Forty-five female rats were randomly assigned to a control group (n = 15) and an experimental group (n = 30) that received injections of gonadotropin releasing hormone antagonist (GnRH a). Causal models were constructed by specifying directed paths between bone traits. The first model tested the hypothesis that the functional relationships between bone traits and body weight were altered by a delay in pubertal onset. GnRH-a injections during puberty altered the covariation between body weight and bone size. The second model was constructed to test the hypothesis that variability in stiffness was causally related to variability in body weight. The model also tested the relationship between the periosteal and endocortical surfaces and their relationship to stiffness. There was no change in the relationship between the surfaces in the GnRH-a group. The third model determined the effect of estradiol on both total area and relative cortical area in both groups. The relationship between periosteal surface and serum estradiol levels was only significant during estrogen suppression. These data suggest that increases in body weight during or prior to puberty may not be protective of bone strength. PMID- 19283427 TI - Short-term exercise in mice increases tibial post-yield mechanical properties while two weeks of latency following exercise increases tissue-level strength. AB - We have previously shown that exercise during growth increases post-yield deformation in C57BL6/129 (B6;129) male tibiae at the expense of reduced pre yield deformation and structural and tissue strength. Other research in the literature indicates that increased mineral content, cross-sectional geometry and structural strength due to exercise can be maintained or increased after exercise ends for as long as 14 weeks. It was therefore hypothesized that after our exercise protocol ended, effects of exercise on mechanical properties would persist, resulting in increased post-yield behavior and rescued strength versus age-matched control mice. Beginning at 8 weeks of age, exercise consisted of running on a treadmill (30 min/day, 12 m/min, 5 degrees incline) for 21 consecutive days. At the end of running and 2 weeks later, in the cortical bone of the tibial mid-diaphyses of B6;129 male mice, changes due to exercise and latency following exercise were assayed by mechanical tests and analyses of cross sectional geometry. Exercise increased structural post-yield deformation compared with weight-matched control mice, without changes in bone size or shape, suggesting that exercised-induced changes in pre-existing bone quality were responsible. Over the 2-week latency period, no growth-related changes were noted in control mice, but exercise-induced changes resulted in increased tissue stiffness and strength versus mice sacrificed immediately after exercise ended. Our data indicate that periods of exercise followed by latency can alter strength, stiffness, and ductility of bone independent of changes in size or shape, suggesting that exercise may be a practical way to increase the quality of the bone extracellular matrix. PMID- 19283428 TI - Giant aneurysm of the common hepatic artery: US and CT imaging findings. AB - Giant hepatic artery aneurysm is a very rare vascular lesion, but can be detected incidentally during abdominal imaging. We report the sonographic and computed tomography (CT) features of a giant hepatic artery aneurysm in a 52-year-old woman presenting with vague abdominal discomfort. This report illustrates that a giant hepatic artery aneurysm can manifest as an incidental large mass in the porta hepatis, and we discuss the role of sonography and CT in the diagnosis of the lesion and review the natural history and clinical presentation of hepatic artery aneurysm. PMID- 19283429 TI - Hepatic adenomas associated with anti-epileptic drugs: a case series and imaging review. AB - We report four patients on long-term antiepileptic drugs, without any history of anabolic steroid or oral contraceptive use, who had path-proven hepatic adenomas at our institution. Imaging review of one case is presented here. An exhaustive literature search reveals only four other such case reports, none from the North American continent. PMID- 19283430 TI - Direct percutaneous embolization of bleeding stomal varices. AB - Stomal variceal bleeding can develop in patients with underlying cirrhosis and portal hypertension. Most patients are best treated with transjugular intrahepatic portosystemic shunt (TIPS) creation because this addresses the underlying problem of portal hypertension. However, some patients are not good candidates for TIPS creation because they have end-stage liver disease or encephalopathy. We describe such a patient who presented with recurrent bleeding stomal varices, which was successfully treated with percutaneous coil embolization. The patient had bleeding-free survival for 1 month before death from unrelated causes. PMID- 19283431 TI - A thermostable metal-tolerant laccase with bioremediation potential from a marine derived fungus. AB - Laccase, an oxidoreductive enzyme, is important in bioremediation. Although marine fungi are potential sources of enzymes for industrial applications, they have been inadequately explored. The fungus MTCC 5159, isolated from decaying mangrove wood and identified as Cerrena unicolor based on the D1/D2 region of 28S and the 18S ribosomal DNA sequence, decolorized several synthetic dyes. Partially purified laccase reduced lignin content from sugarcane bagasse pulp by 36% within 24 h at 30 degrees C. Laccase was the major lignin-degrading enzyme (approximately 24,000 U L(-1)) produced when grown in low-nitrogen medium with half-strength seawater. Three laccases, Lac I, Lac II, and Lac III, of differing molecular masses were produced. Each of these, further resolved into four isozymes by anion exchange chromatography. The N-terminal amino acid sequence of the major isozyme, Lac IId showed 70-85% homology to laccases from basidiomycetes. It contained an N-linked glycan content of 17%. The optimum pH and temperature for Lac IId were 3 and 70 degrees C, respectively, the half-life at 70 degrees C being 90 min. The enzyme was most stable at pH 9 and retained >60% of its activity up to 180 min at 50 degrees C and 60 degrees C. The enzyme was not inhibited by Pb, Fe, Ni, Li, Co, and Cd at 1 mmol. This is the first report on the characterization of thermostable metal-tolerant laccase from a marine-derived fungus with a potential for industrial application. PMID- 19283432 TI - Flexor tendon problems after volar plate fixation of distal radius fractures. AB - Four cases of flexor tendon problems which developed after volar plate fixation of distal radius fractures are presented. All cases were associated with close contact of the screws or distal edge of the plate with the flexor tendons. Poor bone stock or multiple bone fragments allowing loosening of the plate or non locking screws cause the hardware to irritate the flexor tendons and ultimately lead to rupture. The flexor tendons involved include the flexor carpi radialis, flexor pollicis longus and flexor digitorum superficialis, and flexor digitorum profundus to the index and long fingers. PMID- 19283433 TI - Stuck. PMID- 19283435 TI - Utilization of preoperative patient factors to predict postoperative vitamin D deficiency for patients undergoing gastric bypass. AB - INTRODUCTION: Vitamin D deficiency occurring after gastric bypass procedures can predispose patients to calcium and parathyroid hormone (PTH) level abnormalities. The aim of the study is to identify preoperative patient risk factors for postoperative vitamin D deficiency. METHODS: We retrospectively reviewed patients who underwent Roux-en-Y gastric bypass procedures between 2005 and 2006. Patient demographics, laboratory values of calcium, vitamin D, and PTH were followed at quarterly intervals for 1 year. RESULTS: One hundred forty-five patients were included in the study. The mean age for the group was 44 years with an average body mass index of 49.5 kg/m(2). Eighty-six percent of patients were female and 23% was African-American. Forty-two percent of the patients had vitamin D deficiency (<20 ng/mL) either preoperatively or at year 1. The mean calcium levels decreased from 9.39 to 9.16 mg/dL (p < 0.001) while the mean PTH levels increased from 25.7 to 43.9 ng/mL (p < 0.001). A logistic regression model recognized preoperative vitamin D levels, race, and bypass limb length to be the only significant factors (p < 0.05) for postoperative vitamin D deficiency. CONCLUSION: It is important to recognize patients who are at risk for vitamin D deficiency before surgery so that early intervention could be in place to minimize further postoperative deficiency. PMID- 19283436 TI - Cigarette smoking increases the risk for rotator cuff tears. AB - There is little available evidence regarding risk factors for rotator cuff tears. Cigarette smoking may be an important risk factor for rotator cuff disease. The purpose of this study was to determine if cigarette smoking correlates with an increased risk for rotator cuff tears in patients who present with shoulder pain. A questionnaire was administered to 586 consecutive patients 18 years of age or older who had a diagnostic shoulder ultrasound for unilateral, atraumatic shoulder pain with no history of shoulder surgery. Three hundred seventy-five patients had a rotator cuff tear and 211 patients did not. Data regarding cigarette smoking were obtained for 584 of 586 patients. A history of smoking (61.9% versus 48.3%), smoking within the last 10 years (35.2% versus 30.1%), mean duration of smoking (23.4 versus 20.2 years), mean packs per day of smoking (1.25 versus 1.10 packs per day), and mean pack-years of smoking (30.1 versus 22.0) correlated with an increased risk for rotator cuff tear. We observed a dose dependent and time-dependent relationship between smoking and rotator cuff tears. We observed a strong association between smoking and rotator cuff disease. This may indicate smoking is an important risk factor for the development of rotator cuff tears. LEVEL OF EVIDENCE: Level III, prognostic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19283437 TI - Pilates training for use in rehabilitation after total hip and knee arthroplasty: a preliminary report. AB - Recently, a strong emphasis has been placed on establishing rehabilitation protocols after primary total hip and knee arthroplasty in an attempt to shorten, improve, and standardize the postoperative period of recovery. Less invasive surgical techniques, patient demands, and the pressure of insurance regulations have forced postoperative rehabilitation to be placed on an expedited scale. With these concerns in mind, we introduce a pre- and postarthroplasty program involving the Pilates method. Modified exercises have been developed to account for the postoperative precautions and needs of total hip and knee arthroplasty patients. A patient-driven interest in the use of Pilates for postoperative rehabilitation has led to the development of our programs following total hip or knee arthroplasty. In reviewing our early observations of a small series of patients, it appears this technique can be utilized without early complications; however, further studies are necessary to confirm its utility and safety. LEVEL OF EVIDENCE: Level IV, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 19283438 TI - Quantification of fracture healing from radiographs using the maximum callus index. AB - Callus formation and growth are an essential part of secondary fracture healing. Callus growth can be observed radiographically and measured using the "Callus Index," which is defined as the maximum diameter of the callus divided by the diameter of the bone. We compared three groups of patients with tibial fractures treated by external fixation, intramedullary nailing, and casting to assess the validity of using serial measurements of callus index as a measure of fracture healing. When callus index was plotted against time for each patient, the point at which the fracture began to remodel, indicated by the highest point of the curve, was observed as a consistent feature regardless of fixation method. This occurred on average at 2(1/2) weeks after plaster cast removal (14 weeks post injury), 5 weeks after external fixator removal (22 weeks post injury), and 27 weeks post injury for the intramedullary nailed fractures. Because remodeling only occurs once the fracture is stable, a peak in callus index is a reliable sign that the fracture has united. Serial measurements of callus index would therefore appear to offer a simple method of quantifying secondary fracture healing regardless of the treatment method used. LEVEL OF EVIDENCE: Level III, diagnostic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19283439 TI - Ceramic-ceramic bearing decreases osteolysis: a 20-year study versus ceramic polyethylene on the contralateral hip. AB - Although ceramic implants have been in use for many years and they are intended to minimize wear debris it is unknown whether alumina-on-alumina or alumina-on polyethylene produce less wear and osteolysis. We therefore investigated wear and osteolysis on 28 bilateral arthroplasties (one ceramic-ceramic and the contralateral ceramic-polyethylene) of patients who had survived 20 years without revision and without loosening of either hip. Osteolysis was identified on anteroposterior pelvic radiographs and 3-D volume from CT scans. The number of osteolytic lesions detected with CT scan was higher than with radiographs. The number of lesions was higher on the side with the alumina-PE couple. With a similar length of followup on each side, the surface and the volume of osteolysis were consistently higher on the side with the alumina-PE couple. We found no correlation between the volume of osteolysis and the volume of estimated wear in each couple of friction. Hips with osteolysis had a lower Harris score. PMID- 19283440 TI - Synchronous sigmoid and caecal cancers together with a primary renal cell carcinoma. AB - BACKGROUND: Multiple primary neoplasms, a common clinical entity, can be classified as synchronous or metachronous. Renal cell carcinoma, in particular, is associated with a high rate of multiple primary neoplasms. METHODS: We report a case of a 67-year-old Caucasian woman, who was admitted with history of bleeding per rectum. We conducted a literature review of metachronous and synchronous multiple primary neoplasms. RESULTS: Colonoscopy revealed a 3 cm tumour in the caecum and a small sigmoid tubulovillous polyp. Staging computed tomography showed a non-enhancing mass in the upper pole of the right kidney. Following a radical nephrectomy/right hemicolectomy, microscopy demonstrated a renal cell carcinoma. Follow-up colonoscopy visualised a mucosal irregularity at the site of the previous sigmoid polyp, with biopsies confirming invasive adenocarcinoma. Patient underwent a total colectomy with an ileo-rectal anastamosis. CONCLUSION: Multiple primary neoplasms are a well-recognised clinical entity. This report emphasises the need for a comprehensive evidence based work-up in all cancer cases, especially when dealing with renal cell carcinoma, to look for coexisting metachronous/synchronous primary neoplasms. PMID- 19283441 TI - A significantly impaired natural killer cell activity due to a low activity on a per-cell basis in rheumatoid arthritis. AB - To elucidate the characterization of peripheral natural killer (NK) cells in patients with rheumatoid arthritis (RA), we investigated the NK cell activity, the expression of NK cell activating receptors and intracellular molecules. The NK activity was analyzed in 27 RA patients, 22 primary Sjogren's syndrome (SS) patients, and 15 healthy individuals using the (51)Chrominium release assay. The expression of NK cell activating receptors (NKG2D, CD244, CD2, and CD16) and intracellular molecules (granzyme B, perforin, and TCR zeta chain) in CD3-CD56+ cells were characterized by flow cytometry. The serum cytokine levels (IL-6, TNFalpha, and IL-18) were measured using ELISA. Both the NK cell activity and the activity on a per-cell basis were observed to significantly decrease in the RA patients in comparison to the controls. The expression of NKG2D and CD244 also significantly decreased in both the RA and primary SS patients, whereas the significant decrease in the CD16 expression was only observed in the RA patients. The titer of the serum IL-6, TNFalpha, and IL-18 was significantly higher in the RA patients than in the controls. These data suggest that a low NK activity on a per-cell basis might therefore contribute to an impaired NK activity in the patients with RA. PMID- 19283442 TI - Morinda citrifolia Linn. (Rubiaceae) leaf extracts mitigate UVB-induced erythema. AB - Morinda citrifolia Linn. (Rubiaceae) leaves have been used in tropical folk medicine to treat topical inflammation and burns. A carbomer gel base, containing the ethanol extract and juice pressed from the leaves, was evaluated for potential allergenic properties in a repeat-insult patch test in 49 volunteers. To investigate the topical photo-protective properties, the combined ethanol extract and leaf juice were evaluated in a UVB-induced erythema model in 25 volunteers. The crude ethanol extract of M. citrifolia leaves was also evaluated in vitro for potential anti-inflammatory activity in a histamine H-1 receptor antagonism assay. There was no evidence of allergenic potential in the repeat insult patch test. When the combination of ethanol extract and leaf juice was applied, the UVB dose required to induce erythema was almost 3.5 times greater than with untreated skin (P < 0.001). In the histamine H-1 receptor-binding assay, the crude ethanol extract of M. citrifolia leaves inhibited receptor binding by 57%. These results suggest that M. citrifolia leaves are safe for topical use and may be useful in mitigating UVB-induced injury to the skin. PMID- 19283443 TI - Identification of an Arabidopsis unknown small membrane protein targeted to mitochondria, chloroplasts, and peroxisomes. AB - In a functional genomic screen performed by combining an Arabidopsis-yellow fluorescent protein (YFP)-fused complementary DNA (cDNA) library, rat fibroblasts as host and automatic microscopy, we found a short protein with a predictable trans-membrane domain encoded on chromosome 2. In rat fibroblasts, its pattern of distribution was to various organelle-like structures. From the databases, we learned that it has another family member in Arabidopsis and homologs in several other plants, Chlamydomonas and fungi, with a highly conserved N-terminal region. We named this protein from Arabidopsis short membrane protein (SMP) 2. No SMP homologs were found in mammalian sequence databases. When the full-length cDNAs of SMP2 was fused to YFP under the 35S promoter, comparable distribution was observed in Nicotiana benthamiana leaves, suggesting an unknown, evolutionarily conserved localization signal. Similar localization was observed when SMP2 was expressed in N. benthamiana leaves under the control of its own 5' regulatory sequences. Colocalization studies with green fluorescent protein and red fluorescent protein chimeras revealed its colocalization with chloroplasts, peroxisomes, and mitochondria. No localization of SMP2 was observed in the Golgi. Immunostaining with specific antibodies corroborated the SMP2 localization to the three organelles. PMID- 19283444 TI - A new non-natural arginine-like amino acid derivative with a sulfamoyl group in the side-chain. AB - Sulfamoylation of the L-ornithine methyl ester side-chain generates a non-natural arginine isostere which can be coupled with N-Fmoc-L-proline to synthesize analogues which maintain the structural characteristics of the biologically important Pro-Arg dipeptide sequence. As a probe of its biological importance, the sulfamoylated amino acid derivative was also incorporated as P1 residue in tripeptide structures matching the C-terminal subsequence of fibrinogen. The reported results demonstrate that the functionalization of L-ornithine side-chain with a neutral sulfamoyl group can generate an arginine bioisostere which can be used for the synthesis of prototypes of a new class of human thrombin inhibitors. PMID- 19283445 TI - Effect of L-arginine on metabolism of polyamines in rat's brain with extrahepatic cholestasis. AB - Cholestatic encephalopathy results from accumulation of unconjugated bilirubin and hydrophobic bile acids in the brain. The aim of this study was to determine disturbances of polyamine metabolism in the brains of rats with experimental extrahepatic cholestasis and the effects of L-arginine administration. Wister rats were divided into groups: I: sham-operated, II: rats treated with L arginine, III: animals with bile-duct ligation (BDL), and IV: cholestatic-BDL rats treated with L-arginine. Increased plasma gamma-glutamyltransferase and alkaline phosphatase activity and increased bile-acids and bilirubin levels in BDL rats were reduced by administration of L-arginine (P < 0.001). Cholestasis increased the brain's putrescine (P < 0.001) and decreased spermidine and spermine concentration (P < 0.05). The activity of polyamine oxidase was increased (P < 0.001) and diamine oxidase was decreased (P < 0.001) in the brains of BDL rats. Cholestasis increased the activity of arginase (P < 0.05) and decreased the level of citrulline (P < 0.001). Administration of L-arginine in BDL rats prevents metabolic disorders of polyamines and establishes a neuroprotective role in the brain during cholestasis. PMID- 19283446 TI - Women reporting intimate partner violence in India: associations with PTSD and depressive symptoms. AB - Intimate Partner Violence (IPV) is prevalent worldwide and often has mental health sequelae. The aims of this study were (a) to describe the prevalence and the nature of PTSD symptoms among Indian women reporting IPV, (b) to study the relationship between symptoms of PTSD and depression, and (c) to examine the relationship between sexual coercion and PTSD symptoms. Consecutive women (n = 105) presenting to an adult psychiatry outpatient unit of National Institute of Mental Health and Neuro Sciences in South India were recruited. These women were assessed for IPV, sexual coercion, PTSD symptoms, and depression. Fifty-nine women (56%) reported a history of IPV, of whom 41 (70%) also reported sexual coercion. Among women reporting IPV, seven (14%) exceeded cut-off scores for PTSD and twelve (20%) exceeded cut-off scores for sub-threshold PTSD. The majority of those reporting IPV exceeded cut-off scores for a depressive disorder. Compared to women without a history of IPV, women reporting IPV had higher scores on PTSD and depression. Severity of violence and sexual coercion correlated positively (r = 0.39) with PTSD severity. The findings highlight the importance of screening women for IPV and its sequelae, in mental health settings. PMID- 19283447 TI - Novel quantitative echocardiographic parameters in acute PE. AB - BACKGROUND: The tricuspid annular plane systolic excursion (TAPSE) and the right ventricular performance index (RVPI) are quantitative measurements that are valid predictors of clinical outcomes in CHF, MI, PAH, and chronic pulmonary disease. We sought to measure TAPSE and RVPI in patients diagnosed with acute pulmonary embolism (APE) to assess for correlation with known predictors of clinical outcomes. METHODS: Patients admitted with APE had echocardiograms performed within 24 h of diagnosis and B-type natriuretic peptide (BNP) drawn on admission. Serial troponins were measured for the first 48 h of the hospital stay, and clinical course was followed until discharge. RESULTS: A total of 29 patients were enrolled in the study. Compared to those with a normal study, significantly more patients with an abnormal TAPSE had an elevated BNP (60% vs. 5%; P = 0.004) and troponin (50% vs. 11.1%; P = 0.042). The mean TAPSE was 22.3 mm when BNP was normal and 17.4 mm when elevated (P = 0.003). TAPSE values were significantly lower in patients with abnormal RV function by echocardiogram graded by a blinded cardiologist (17.6 mm vs. 21.7 mm; P = 0.03). Both TAPSE and RVPI correlated significantly with septal flattening, RVEDD, and RVEDD/LVEDD by echo. CONCLUSIONS: TAPSE has good correlation with surrogate markers for morbidity and mortality in APE, and both TAPSE and RVPI seem to perform as well as the standard echo parameters used to assess RV function. Both are objective and easy to measure, and therefore warrant prospective study in larger patient groups, with assessment of clinical outcomes. PMID- 19283448 TI - Genetic polymorphisms involved in folate metabolism and concentrations of methylmalonic acid and folate on plasma homocysteine and risk of coronary artery disease. AB - OBJECTIVES: Alterations in the enzymes involved in homocysteine (Hcy) metabolism or vitamin deficiency could play a role in coronary artery disease (CAD) development. This study investigated the influence of MTHFR and MTR gene polymorphisms, plasma folate and MMA on Hcy concentrations and CAD development. MMA and folate concentrations were also investigated according to the polymorphisms. METHODS: Two hundred and eighty-three unrelated Caucasian individuals undergoing coronary angiography (175 with CAD and 108 non-CAD) were assessed in a case-control study. Plasma Hcy and MMA were measured by liquid chromatography/tandem mass spectrometry. Plasma folate was measured by competitive immunoassay. Dietary intake was evaluated using a nutritional questionnaire. Polymorphisms MTHFR and MTR were investigated by polymerase chain reaction (PCR) followed by enzyme digestion or allele-specific PCR. RESULTS: Hcy mean concentrations were higher in CAD patients compared to controls, but below statistical significance (P = 0.246). Increased MMA mean concentrations were frequently observed in the CAD group (P = 0.048). Individuals with MMA concentrations >0.5 micromol/l (vitamin B(12) deficiency) were found only in the CAD group (P = 0.004). A positive correlation between MMA and Hcy mean concentrations was observed in both groups, CAD (P = 0.001) and non-CAD (P = 0.020). MMA mean concentrations were significantly higher in patients with hyperhomocysteinemia in both groups, CAD and non-CAD (P = 0.0063 and P = 0.013, respectively). Folate mean concentration was significantly lower in carriers of the wild-type MTHFR 1298AA genotype (P = 0.010). CONCLUSION: Our results suggest a correlation between the MTHFR A1298C polymorphism and plasma folate concentration. Vitamin B(12) deficiency, reflected by increased MMA concentration, is an important risk factor for the development both of hyperhomocysteinemia and CAD. PMID- 19283449 TI - Enoxaparin dosing in the elderly using adjusted body weight. AB - We prospectively compared anti-Xa activity in 61 elderly (>65 years) subjects receiving enoxaparin according to standard or adjusted body weight (ABW) dosing. In the ABW dosing group, mean patient age was 76 years, mean weight 80 kg, mean serum creatinine 1.0 mg/dl, and mean CrCl 48 ml/min. ABW dosing resulted in 57% of elderly study subjects achieving anti-Xa activity of 0.5-1.0 IU/ml, and 80% achieving anti-Xa activity of 0.5-1.2 IU/ml. Compared to standard dosing, for all subjects ABW dosing of enoxaparin was associated with a more favorable mean anti Xa activity (0.98 IU/ml vs. 1.28 IU/ml, P = 0.001), fewer highest-risk (>1.5 IU/ml) supratherapeutic anti-Xa levels (0% vs. 28%, P = 0.001), and more frequent therapeutic levels among women (64% vs. 25%, P = 0.001). ABW dosing of enoxaparin may be beneficial in elderly patients aged 65 and older, and its benefit appears to be more pronounced in female patients. PMID- 19283451 TI - The association of a mild FXII deficiency with myocardial infarction and venous thrombosis is completely unjustified. PMID- 19283452 TI - Heavy ion irradiation increases apoptosis and STAT-3 expression, led to the cells arrested at G2/M phase in human hepatoma SMMC-7721 cells. AB - BACKGROUND: The impact of STAT-3 expression on the apoptosis of human hepatomas cell SMMC-7721 line induced by X-ray and carbon ion irradiations was investigated. METHODS: Human hepatoma SMMC-7721 cells were irradiated with a carbon ion beam and X-ray. Cell survival was determined by a standard colony forming assay. STAT-3 protein expression was analysed by Western Immunoblots. Cell cycle and apoptosis were performed by flow cytometry. RESULTS: The viability of SMMC-7721 cells decreased with increasing dose of the carbon ion beam, and the high-LET carbon ion beam led to the cells getting arrested at G(2)/M phase. Western Blot analyses show that STAT-3 expression increased with increasing radiation dose. The carbon ion irradiation induced cell apoptosis and significantly promoted the expression of STAT-3 gene compared with the X-ray irradiation. The apoptosis rate is correlated with the expression of STAT-3 in human hepatoma SMMC-7721 cells after exposure to different doses of X-ray and heavy ion beam. CONCLUSIONS: Heavy ion irradiation increases the expression of STAT-3 gene, makes SMMC-7721 cells arrested at G(2)/M phase and increases cell apoptosis in comparison with that induced by low-LET X-ray. The STAT-3 expression may be regarded as a protected reaction when the cancerous cells suffer a strong stimulus such as high-LET irradiation. The interaction of STAT-3 expression and other cytokines in human hepatoma and the relationship between STAT-3 and radiation-induced apoptosis remain to be clarified in the future. PMID- 19283453 TI - Poly (lactic-co-glycolic acid) as a controlled release delivery device. AB - Poly (lactic-co-glycolic acid) (PLGA) is a biodegradable polymer used to make resorbable sutures, and is also used in other applications in tissue engineering. Being an artificial polymer, its degradation rate can be tailored to suit its application. It can be easily moulded into structures with suitable mechanical strength and degrades into relatively harmless products in the body. Its adjustable degradation rate also makes it a potentially excellent controlled release delivery device. However, the functionalization of PLGA with bioactive molecules usually requires extensive chemical modification. Chemical modification may compromise the mechanical strength of PLGA and inactivate the bioactive molecules. In this paper, a study is done to investigate the coating of an angiogenic factor on unmodified PLGA suture substrates for the differentiation of human mesenchymal stem cells (hMSC) into endothelial cells (EC). The results show that the method used to anchor vascular endothelial growth factor (VEGF) onto the PLGA surface can enable the gradual release of VEGF from the substrate into solution to induce the differentiation of hMSCs into ECs. Thus, this method can potentially be used to coat PLGA materials like sutures, meshes and scaffolds, rendering them functional as effective controlled release delivery devices for a wide range of bioactive molecules. PMID- 19283454 TI - Comparison of failure mechanisms for cements used in skeletal luting applications. AB - Glass Polyalkenoate Cements (GPCs) based on strontium calcium zinc silicate (Sr Ca-Zn-SiO(2)) glasses and low molecular weight poly(acrylic acid) (PAA) have been shown to exhibit suitable compressive strength (65 MPa) and flexural strength (14 MPa) for orthopaedic luting applications. In this study, two such GPC formulations, alongside two commercial cements (Simplex P and Hydroset) were examined. Fracture toughness and tensile bond strength to sintered hydroxyapatite and a biomedical titanium alloy were examined. Fracture toughness of the commercial Poly(methyl methacrylate) cement, Simplex P, (3.02 MPa m(1/2)) was superior to that of the novel GPC (0.36 MPa m(1/2)) and the commercial calcium phosphate cement, Hydroset, for which no significant fracture toughness was obtained. However, tensile bond strengths of the novel GPCs (0.38 MPa), after a prolonged period (30 days), were observed to be superior to commercial controls (Simplex P: 0.07 MPa, Hydroset: 0.16 MPa). PMID- 19283456 TI - Accurate potentiometric determination of lipid membrane-water partition coefficients and apparent dissociation constants of ionizable drugs: electrostatic corrections. AB - PURPOSE: Potentiometric lipid membrane-water partition coefficient studies neglect electrostatic interactions to date; this leads to incorrect results. We herein show how to account properly for such interactions in potentiometric data analysis. MATERIALS AND METHODS: We conducted potentiometric titration experiments to determine lipid membrane-water partition coefficients of four illustrative drugs, bupivacaine, diclofenac, ketoprofen and terbinafine. We then analyzed the results conventionally and with an improved analytical approach that considers Coulombic electrostatic interactions. RESULTS: The new analytical approach delivers robust partition coefficient values. In contrast, the conventional data analysis yields apparent partition coefficients of the ionized drug forms that depend on experimental conditions (mainly the lipid-drug ratio and the bulk ionic strength). This is due to changing electrostatic effects originating either from bound drug and/or lipid charges. A membrane comprising 10 mol-% mono-charged molecules in a 150 mM (monovalent) electrolyte solution yields results that differ by a factor of 4 from uncharged membranes results. CONCLUSION: Allowance for the Coulombic electrostatic interactions is a prerequisite for accurate and reliable determination of lipid membrane-water partition coefficients of ionizable drugs from potentiometric titration data. The same conclusion applies to all analytical methods involving drug binding to a surface. PMID- 19283457 TI - Young adult gambling behaviors and their relationship with the persistence of ADHD. AB - Young adulthood is a period renowned for engagement in impulsive and risky behaviors, including gambling. There are some indications that young adults exhibit higher gambling rates in comparison to older adults. Problem gambling has also been linked to ADHD. This longitudinal study examines the relationship between gambling and ADHD among an epidemiological sample of young adults (n = 235; males = 179, females = 56) aged 18-24. Results indicate that individuals who report childhood ADHD symptoms which persist into young adulthood experience greater gambling problem severity than participants with no ADHD or those with non-persistent ADHD. PMID- 19283458 TI - Conflict nightmares and trauma in Aceh. AB - In both the Acehnese and Indonesian languages, there is no single lexical term for "nightmare." And yet findings from a large field research project in Aceh that examined post traumatic experience during Aceh's nearly 30-year rebellion against the Indonesian state and current mental distress revealed a rich variety of dream narratives that connect directly and indirectly to respondents' past traumatic experiences. The results reported below suggest that even in a society that has a very different cultural ideology about dreams, where "nightmares" as such are not considered dreams but rather the work of mischievous spirits called jin, they are still a significant part of the trauma process. We argue that it is productive to distinguish between terrifying and repetitive dreams that recreate the traumatic moment and the more ordinary varieties of dreams that Acehnese reported to their interviewers. Nightmares that refer back to conflict events do not appear as an elaborated feature of trauma as the condition is understood by people in Aceh, but when asked further about their dreams, respondents who reported symptoms suggestive of PTSD were more likely to report PTSD-like dreams, memory intrusions that repeat the political violence of the past. PMID- 19283459 TI - Interatrial right-to-left conduction in patients with paroxysmal atrial fibrillation. AB - PURPOSE: We wanted to illustrate the right-to-left impulse propagation routes during sinus in patients with paroxysmal atrial fibrillation (PAF), as alterations in conduction patterns have been linked to the pathogenesis of PAF, and as no large patient materials have been published. METHODS: Patients underwent 3-D electroanatomical contact mapping prior to catheter ablation. The site of the earliest left atrial (LA) activation was determined. RESULTS: Three different interatrial routes were identified, either as solitary pathways (36/50 patients, 72%) or in their combinations (14/50). Bachmann's bundle (BB) was involved in the majority of the cases with solitary routes (25/36). More seldom, impulse propagation occurred near the oval fossa (FO) (7/36) or the coronary sinus ostium (4/36). In patients with combined routes, both the BB (10/14) and FO routes (11/14) were included in most cases. CONCLUSIONS: In PAF patients, LA can be activated during sinus rhythm through three distinct connections, either encompassing a single route or via any combination of these connections. In one third, the earliest LA activation occurs outside BB. The knowledge of the propagation patterns may give insight into the pathophysiology of PAF and into refining ablation therapy. PMID- 19283460 TI - Atrial fibrillation. Introduction. PMID- 19283461 TI - Brief report: sensory abnormalities as distinguishing symptoms of autism spectrum disorders in young children. AB - The purpose of this study was to explore the sensory profile of young children with ASD compared to young children with other developmental delays (DD) at first ASD assessment. Results found that young children with ASD had more tactile and taste/smell sensitivities and difficulties with auditory filtering than young children with other DD. Moreover, sensory scores were significantly correlated with stereotyped interests and behaviors. These findings support the hypotheses that young children with ASD show more sensory impairments than young children with other DD and that sensory symptoms are significantly related to stereotyped interests and behaviors. Results also suggest that sensory abnormalities are distinguishing symptoms of ASD that should be considered in diagnostic algorithms for younger cohorts. PMID- 19283462 TI - Sequence signatures in envelope protein may determine whether flaviviruses produce hemorrhagic or encephalitic syndromes. AB - We analyzed the envelope proteins in pathogenic flaviviruses to determine whether there are sequence signatures associated with the tendency of viruses to produce hemorrhagic disease (H-viruses) or encephalitis (E-viruses). We found that, at the position corresponding to the glycosylated Asn-67 in dengue virus, asparagine (Asn) occurs in all seven viral species that cause hemorrhagic disease in humans. Furthermore, Asn was extremely rare at position 67 in six flaviviruses that cause encephalitis, being replaced by Asp in four of them. Of the 3,246 sequences from H- and E-viruses, we found that 2,916 sequences (90%) contained Asn in position 67 for H-viruses or Asp in position 67 for E-viruses. The change from Asn-67 that is prevalent in H-viruses to Asp-67 (common in E-viruses) contributes to a stronger electrostatically negative surface in the E-viruses as compared to the H viruses. These findings should help predicting the disease potential of emerging and re-emerging flaviviruses and understanding the relationship between protein structure and disease outcome. PMID- 19283463 TI - Investigation of genetically mediated child effects on maltreatment. AB - Theory and empirical evidence suggest that children's genetically influenced characteristics help to shape the environments they experience, including the parenting they 'receive'. The extent of these genetically-mediated child effects on childhood maltreatment is not well known. The present study estimates the magnitude of genetically mediated child effects on maltreatment in 3,297 twins and siblings who were part of a large nationally representative sample of adolescents (ADD health). Participants in early adulthood retrospectively reported their experiences of physical and sexual maltreatment and neglect. Results are consistent with small genetically-mediated child effects on physical maltreatment and neglect, and none on sexual maltreatment, and all three forms of maltreatment are influenced mainly by idiosyncratic individual circumstances. PMID- 19283464 TI - Transportation costs impede sustained adherence and access to HAART in a clinic population in southwestern Uganda: a qualitative study. AB - The cost of transportation for monthly clinic visits has been identified as a potential barrier to antiretroviral (ARV) adherence in sub-Saharan Africa and elsewhere, although there is limited data on this issue. We conducted open-ended interviews with 41 individuals living with HIV/AIDS and attending a clinic in Mbarara, Uganda, to understand structural barriers to ARV adherence and clinical care. Almost all respondents cited the need to locate funds for the monthly clinic visit as a constant source of stress and anxiety, and lack of money for transportation was a key factor in cases of missed doses and missed medical appointments. Participants struggled with competing demands between transport costs and other necessities such as food, housing and school fees. Our findings suggest that transportation costs can compromise both ARV adherence and access to care. Interventions that address this barrier will be important to ensure the success of ARV programs in sub-Saharan Africa. PMID- 19283465 TI - Social information processing skills in children with histories of heavy prenatal alcohol exposure. AB - Based on caregiver report, children with prenatal alcohol exposure have difficulty with social functioning, but little is known about their social cognition. The current study assessed the social information processing patterns of school-age children with heavy prenatal alcohol exposure using a paradigm based on Crick and Dodge's reformulated six-stage model. Fifty-two children (aged 7-11) with and without heavy prenatal alcohol exposure were tested using a structured interview measure of social information processing involving 18 videotaped vignettes of children in group entry and provocation situations. Alcohol-exposed children displayed maladaptive processing patterns on the goal, response generation, and response evaluation steps in group entry situations, and encoding, attribution, response evaluation, and enactment steps during provocation situations. Children with heavy prenatal alcohol exposure also had difficulty on the Test of Problem Solving, and performance correlated with social information processing measures. Such difficulties may lead to problems in social functioning and warrant early intervention. PMID- 19283466 TI - Effect of liver ischemia-reperfusion injury on the activity of neurons in the rat brain. AB - Liver ischemia-reperfusion injury (LIRI) influences different body cells. Little is known about the effect of LIRI on the activity of neurons. Response of neurons to: (1) single ligation of hepatic artery (LIRIa) for 30 min and (2) combined ligation of portal triade (common hepatic artery, portal vein, common bile duct, LIRIb) for 15 min was investigated in Wistar rats. Ninety minutes, 5 h, and 24 h after liver reperfusion, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), interleukin 1alpha (IL-1alpha), and tumor necrosis factor alpha (TNFalpha) serum levels were analyzed and Fos-immunolabeled cells counted in subfornical organ (SFO), suprachiasmatic (SCH), paraventricular (PVN), supraoptic (SON), arcuate (ARC), and ventromedial (VMN) hypothalamic nuclei, locus coeruleus (LC), nucleus of the solitary tract (NTS), and A1/C1 catecholaminergic cell groups. LIRIb increased ALT serum level after 90 min and 24 h while AST activity only after 24 h in all experimental groups. IL-1alpha serum level was increased only after 90 min of LIRIb while TNFalpha level did not change. Ninety minutes after surgeries more Fos-immunostained cells occurred in both LIRIs than sham-operated animals in all structures studied. More distinct Fos expression occurred after LIRIb than LIRIa in SON, PVN, VMN, and NTS. Five hours after both LIRIs, Fos increased in the parabrachial nucleus (PBN) and NTS. Twenty-four hours after both LIRIs Fos incidence decreased in all groups. Although the present data indicate that increased neuronal activity after both LIRIs is mainly a consequence of the liver damage itself partial impact of non specific factors can not be excluded. However, the anatomical distribution of Fos occurrence detected after LIRIs gives great opportunity to perform a targeted phenotypic identification of the activated neurons by LIRIs in the subsequent experiments. PMID- 19283467 TI - Improved behavioral response as a valid biomarker for drug screening program in transgenic rodent models of tauopathies. AB - Neurodegenerative tauopathies are defined as a group of dementia and movement disorders characterized by prominent filamentous tau inclusions and degeneration located within certain brain regions. Their common sign is a presence of proteinaceous aggregates composed of hyperphosphorylated and truncated tau proteins. The molecular mechanisms of the disease still remain unresolved, therefore transgenic organisms displaying tau-related neurodegenerative cascade have been created to allow decoding of individual pathways involved in human pathological conditions. Moreover, use of transgenic model organisms enables the application of potential therapeutic approaches. The expression of mutated or misfolded tau as a transgene in vivo leads to significant alteration of neurobehavioral features of experimental animal, therefore detailed classification of behavioral phenotype become one of the first crucial analyses, while it functionally correlates with central nervous system impairment. Currently, two major types of behavioral impairment have been described in transgenic rodent models of tauopathies, (1) progressive motor impairment associated with muscular weakness and premature death and (2) age-related impairment of cognitive functions attended with unaffected motor status. Up to the present, only transgenic models displaying motor impairment were successfully applied into the drug trials targeting misfolded tau protein, despite their behavioral inconsistence with clinical profile of progressive human tauopathy. The aim of this study was, therefore, to summarize the pros and cons of used transgenic rodent models mimicking human tauopathies in connection with development of therapeutic strategies. PMID- 19283468 TI - Molecular mechanisms leading to neuroprotection/ischemic tolerance: effect of preconditioning on the stress reaction of endoplasmic reticulum. AB - Ischemic tolerance can be developed by prior ischemic non-injurious stimulus preconditioning. The molecular mechanisms underlying ischemic tolerance are not yet fully understood. The purpose of this study is to evaluate the effect of preconditioning/preischemia on ischemic brain injury. We examined the endoplasmic reticulum stress response (unfolded protein response (UPR)) by measuring the mRNA and protein levels of specific genes such as ATF6, GRP78, and XBP1 after 15 min 4 VO ischemia and different times of reperfusion (1, 3, and 24 h). The data from the group of naive ischemic rats were compared with data from the group of preconditioned animals. The results of the experiments showed significant changes in the gene expression at the mRNA level in the all ischemic/reperfusion phases. The influence of preischemia on protein level of XBP was significant in later ischemic times and at 3 h, the reperfusion reached 230% of the controls. The protein levels of GRP78 in preischemic animals showed a significant increase in ischemic and reperfusion times. They exceeded to 50% levels of corresponding naive ischemic/reperfusion groups. Preconditioning also induced remarkable changes in the levels of ATF6 protein in the ischemic phase (about 170%). The levels of ATF6 remained elevated in earlier reperfusion times (37 and 62%, respectively) and persisted significantly elevated after 24 h of reperfusion. This data suggest that preconditioning paradigm (preischemia) underlies its neuroprotective effect by the attenuation of ER stress response after acute ischemic/reperfusion insult. PMID- 19283469 TI - Differential regulation of CuZnSOD expression in rat brain by acute and/or chronic stress. AB - Neuroendocrine stress (NES) causes increase of glucocorticoids and alters physiological levels of reactive oxygen species production in cells, which might involve modifications in the antioxidant defense system. We investigated the hypothesis that acute, chronic, or combined stress alters copper-zinc superoxide dismutase (CuZnSOD) expression pattern at both, mRNA and subcellular protein level in the cerebral cortex and hippocampus of rats and that there may be a relationship between stress-induced corticosterone and CuZnSOD expression. The most effective stress model which led to the most pronounced changes in CuZnSOD expression patterns was also investigated. Our results demonstrated that acute stress immobilization up-regulates mRNA expression of hippocampal CuZnSOD, while cytosolic protein expression of this enzyme was increased in both brain structures. Chronic stress isolation had no effect on either mRNA and protein expression level and caused a lack of significant up-regulation to a novel acute stressors. The presence of this protein in nuclear fractions of both brain structures was also confirmed. The elevated cytosolic CuZnSOD protein levels following acute immobilization might reflect on the defense system against oxidative stress. Chronic isolation compromises CuZnSOD protein expression, which may lead to the inefficient defense against reactive oxygen species (ROS). The stress-triggered CuZnSOD protein expression was not correlated by the corresponding mRNA. The results suggest that different stress models exert a different degree of influence on mRNA and protein level of CuZnSOD in both brain structures as well as serum corticosterone. PMID- 19283470 TI - Ischemia-induced mitochondrial apoptosis is significantly attenuated by ischemic preconditioning. AB - Ischemic preconditioning (IPC) represents an important adaptation of CNS to sub lethal ischemia, which results in increased tolerance of CNS to the lethal ischemia. Ischemia-induced mitochondrial apoptosis is considered to be an important event leading to neuronal cell death after cerebral blood flow arrest. In presented study, we have determined the effect of IPC on ischemia/reperfusion induced mitochondrial apoptosis. Global brain ischemia was induced by permanent occlusion of vertebral arteries and temporal occlusion of carotid arteries for 15 min. Rats were preconditioned by 5 min of sub-lethal ischemia and 2 days later 15 min of lethal ischemia was induced. With respect to mitochondrial apoptosis initiation, translocation of p53 to mitochondria was observed in hippocampus but not in cerebral cortex. However, level of both apoptotic bax and anti-apoptotic bcl-xl in both hippocampal and cortical mitochondria was unchanged after global brain ischemia. Detection of genomic DNA fragmentation as well as Fluoro-Jade C staining showed that ischemia induces apoptosis in vulnerable CA1 layer of rat hippocampus. IPC abolished completely ischemia-induced translocation of p53 to mitochondria and had significant protective effect on ischemia-induced DNA fragmentation. In addition, significant decrease of Fluoro-Jade C positive cells was observed as well. Our results indicate that IPC abolished almost completely both initiation and execution of mitochondrial apoptosis induced by global brain ischemia. PMID- 19283471 TI - A genetic strategy involving a glycosyltransferase promoter and a lipid translocating enzyme to eliminate cancer cells. AB - The most common therapeutic strategy for the treatment of cancer uses antimetabolites, which block uncontrolled division of cancer cells and kill them. However, such antimetabolites also kill normal cells, thus yielding detrimental side effects. This emphasizes the need for an alternative therapy, which would have little or no side effects. Our approach involves designing genetic means to alter surface lipid determinants that induce phagocytosis of cancer cells. The specific target of this strategy has been the enzyme activity termed aminophospholipid translocase (APLT) or flippase that causes translocation of phosphatidylserine (PS) from the outer to the inner leaflet of the plasma membrane in viable cells. Efforts to identify the enigmatic, plasma membrane APLT of mammalian cells have led investigators to some P-type ATPases, which have often proven to be the APLT of internal membranes rather than the plasma membrane. By measuring kinetic parameters for the plasma membrane APLT activity, we have shown that the P-type ATPase Atp8a1 is the plasma membrane APLT of the tumorigenic N18 cells, but not the non-tumorigenic HN2 (hippocampal neuron x N18) cells. Targeted knockdown of this enzyme causes PS externalization in the N18 cells, which would trigger phagocytic removal of these cells. But how would we specifically express the mutants or antisense Atp8a1 in the cancer cells? This has brought us to a glycosyltransferase, GnT-V, which is highly expressed in the transformed cells. By using the GnT-V promoter to drive a luciferase reporter gene we have demonstrated a dramatic increase in luciferase expression selectively in tumor cells. The described strategy could be tested for the removal of cancer cells without the use of antimetabolites that often kill normal cells. PMID- 19283472 TI - A test battery approach to the ecotoxicological evaluation of cadmium and copper employing a battery of marine bioassays. AB - Heavy metals are ubiquitous contaminants of the marine environment and can accumulate and persist in sediments. The toxicity of metal contaminants in sediments to organisms is dependent on the bioavailability of the metals in both the water and sediment phases and the sensitivity of the organism to the metal exposure. This study investigated the effects of two metal contaminants of concern (CdCl(2) and CuCl(2)) on a battery of marine bioassays employed for sediment assessment. Cadmium, a known carcinogen and widespread marine pollutant, was found to be the least toxic of the two assayed metals in all in vivo tests. However, CdCl(2) was found to be more toxic to the fish cell lines PLHC-1 and RTG 2 than CuCl(2). Tisbe battagliai was the most sensitive species to both metals and the Microtox and cell lines were the least sensitive (cadmium was found to be three orders of magnitude less toxic to Vibrio fischeri than to T. battagliai). The sensitivity of Tetraselmis suecica to the two metals varied greatly. Marine microalgae are among the organisms that can tolerate higher levels of cadmium. This hypothesis is demonstrated in this study where it was not possible to derive an EC(50) value for CdCl(2) and the marine prasinophyte, T. suecica. Conversely, CuCl(2) was observed to be highly toxic to the marine alga, EC(50) of 1.19 mg l( 1). The genotoxic effect of Cu on the marine phytoplankton was evaluated using the Comet assay. Copper concentrations ranging from 0.25 to 2.50 mg l(-1) were used to evaluate the effects. DNA damage was measured as percent number of comets and normal cells. There was no significant DNA damage observed at any concentration of CuCl(2) tested and no correlation with growth inhibition and genetic damage was found. PMID- 19283473 TI - Interactions between chronic stress and chronic consumption of caffeine on the enzymatic antioxidant system. AB - We studied the effect of chronic caffeine on parameters related to oxidative stress in different brain regions of stressed and non-stressed rats. Wistar rats were divided into three groups: control (receiving water), caffeine 0.3 g/L and caffeine 1.0 g/L (in the drinking water). These groups were subdivided into non stressed and stressed (repeated restraint stress during 40 days). Lipid peroxide levels and the total radical-trapping potential were assessed, as well as antioxidant enzyme activities superoxide dismutase, gluthatione peroxidase, and catalase in hippocampus, striatum and cerebral cortex. Results showed interactions between stress and caffeine, especially in the cerebral cortex, since caffeine increased the activity of some antioxidant enzymes, but not in stressed animals. We concluded that chronic administration of caffeine led, in some cases, to increased activity of antioxidant enzymes. However, these effects were not observed in the stressed animals. PMID- 19283474 TI - Cortical NMDA receptor expression in human chronic alcoholism: influence of the TaqIA allele of ANKK1. AB - Real-time RT-PCR normalized to GAPDH was used to assay N-methyl-D-aspartate (NMDA) receptor NR1, NR2A and NR2B subunit mRNA in human autopsy cortex tissue from chronic alcoholics with and without comorbid cirrhosis of the liver and matched controls. Subunit expression was influenced by the subject's genotype. The TaqIA polymorphism selectively modulated NMDA receptor mean transcript expression in cirrhotic-alcoholic superior frontal cortex, in diametrically opposite ways in male and female subjects. Genetic make-up may differentially influence vulnerability to brain damage by altering the excitation: inhibition balance, particularly in alcoholics with comorbid cirrhosis of the liver. The TaqIA polymorphism occurs within the poorly characterised ankyrin-repeat containing kinase 1 (ANKK1) gene. Using PCR, ANKK1 mRNA transcript was detected in inferior temporal, occipital, superior frontal and primary motor cortex of control human brain. ANKK1 expression may mediate the influence of the TaqIA polymorphism on phenotype. PMID- 19283475 TI - Comparative effects of acute or chronic administration of levodopa to 6-OHDA lesioned rats on the expression and phosphorylation of N-methyl-D-aspartate receptor NR1 subunits in the striatum. AB - N-methyl-D-aspartate receptor (NMDA) has been increasingly implicated in the formation and maintenance of various forms of behavioral and synaptic plasticity. Recent evidence has linked striatal NMDA function to the adverse effects of long term dopaminergic treatment in Parkinson's disease. The subcellular distribution and phosphorylation of NMDA subunit, NR1, reflects NMDA receptor activity. To elucidate molecular mechanisms that underlie the persisting alterations in motor response occurring with levodopa treatment of parkinsonian patients, we evaluated the effects of unilateral nigrostriatal depletion with 6-hydroxydopamine and subsequent levodopa treatment on motor responses and NR1 alterations. Three weeks of levodopa administration to rats shortened the rotational duration and increased the peak turning responses, which lasted after withdrawal of chronic levodopa treatment. We found a significant reduction in the abundance of both phosphorylated NR1 on serine residues 890 and 896 (pNR1S890 and pNR1S896) and NR1 in the cell plasma membrane of lesioned striatum. Chronic treatment of lesioned rats with levodopa markedly upregulated pNR1S890, pNR1S896, and pNR1S897 in lesioned striatum with a concomitant normalization of the plasma membrane NR1 abundance. The magnitude of increased pNR1S890, pNR1S896, and pNR1S897 is dependent on the number of levodopa injections and is paralleled by a sensitization of the rotational response. Our data indicate that glutamate signaling is triggered during the levodopa administration. Activated NMDA receptor NR1-mediated mechanisms are involved in the persistent expression of the motor response alterations that appear during chronic levodopa therapy of parkinsonian rats and continue after treatment withdrawal. PMID- 19283476 TI - Spectral energy of ECG morphologic differences to predict death. AB - Unstable conduction system bifurcations following ischemia and infarction are associated with variations in the electrocardiographic activity spanning the heart beat. In this paper, we investigate a spectral energy measure of morphologic differences (SE-MD) that quantifies aspects of these changes. Our measure uses a dynamic time-warping approach to compute the time-aligned morphology differences between pairs of successive sinus beats in an electrocardiographic signal. While comparing beats, the entire heart beat signal is analyzed in order to capture changes affecting both depolarization and repolarization. We show that variations in electrocardiographic activity associated with death can be distinguished by their spectral characteristics. We developed the SE-MD metric on holter data from 764 patients from the TIMI DISPERSE2 dataset and tested it on 600 patients from the TIMI MERLIN dataset. In the test population, high SE-MD was strongly associated with death over a 90 day period following non-ST-elevation acute coronary syndrome (HR 10.45, p < 0.001) and showed significant discriminative ability (c-statistic 0.85). In comparison with heart rate variability and deceleration capacity, SE-MD was also the most significant predictor of death in the study population. Furthermore, SE-MD had low correlation with these other measures, suggesting that complementary use of the risk variables may allow for more complete assessment of cardiac health. PMID- 19283477 TI - Recent trends for colorectal cancer screening in HIV-infected patients. AB - We planned to investigate the recent trends for colorectal cancer (CRC) screening in human immunodeficiency virus (HIV) as compared to non-HIV by interviewing consecutive patients in outpatient clinics during September 2007. Out of a total of 300 patients interviewed, 205 met the inclusion criteria. One hundred and fourteen were HIV-infected, while 91 were non-HIV. HIV received more office visits per year. Despite the high incidence of adenomas on screening colonoscopy, HIV patients were less likely to undergo any type of CRC screening test. A higher number (>or=10) of annual clinic visits and the presence of co-morbid conditions were associated with being up-to-date for CRC screening in HIV patients. Conclusion CRC screening is underutilized in HIV patients. Multiple office visits per year are associated with being up-to-date for CRC screening. Hence, increasing patients' compliance with office visits and educating them about the importance of CRC screening may indirectly increase compliance with CRC screening. PMID- 19283478 TI - Development of a novel esophageal stricture simulation. AB - BACKGROUND AND STUDY AIMS: Esophageal stricture dilatation has a significant morbidity and mortality and training can be difficult to obtain. The aim of the study was to investigate the face validity of a novel stricture simulation and evaluate its utility for training in balloon-dilatation technique. METHODS: Single-use stricture simulations were used to adapt a mechanical model for use in esophageal stricture dilatation. Face validity was evaluated using a questionnaire survey following a 40-min hands-on training session. Performance improvement was evaluated as part of a randomized blinded controlled trial. RESULTS: Face validity was established, with all trainees and instructors rating it as good or excellent overall and as adequately realistic or better in appearance. About 74% found it fairly or very realistic to dilate and 91% found it fairly or very useful for learning balloon-dilatation technique. Significant improvements in performance compared with controls were found following use of the simulation in a training episode. CONCLUSIONS: The novel esophageal stricture simulation had good face validity and has been shown to improve performance when used for training in balloon-dilatation technique. Its use allows practice without risk to patients or the need for animal cadavers. PMID- 19283479 TI - Blood flow in a compliant vessel by the immersed boundary method. AB - In this paper we develop a computational approach to analyze hemodynamics in the aorta; this may serve as a useful tool in the development of noninvasive methods to detect early onset of diseases such as aneurysms and stenosis in major blood vessels. We introduce a mathematical model which describes the interaction of blood flow with the aortic wall; this model is based on the immersed boundary method. A two-dimensional vessel model is constructed, the velocity at the inlet is prescribed based on the information from the Magnetic Resonance Imaging data measured in the aorta of a healthy subject, and the velocity at the outlet is prescribed by driving the pressure level reproduced from the literature. The mathematical model is validated by comparing with well-known solutions of the viscous incompressible Navier-Stokes equations, i.e., Womersley flow. The hysteresis behavior in the pressure-diameter relation is observed when the viscoelastic material property of the arterial wall is taken into consideration. Five different shapes of aortic wall are considered for comparison of the flow patterns inside the aorta: one for the normal aorta, two for the dilated aorta, and two for the constrictive aorta. PMID- 19283480 TI - A model of electrical conduction in cardiac tissue including fibroblasts. AB - Fibroblasts are abundant in cardiac tissue. Experimental studies suggested that fibroblasts are electrically coupled to myocytes and this coupling can impact cardiac electrophysiology. In this work, we present a novel approach for mathematical modeling of electrical conduction in cardiac tissue composed of myocytes, fibroblasts, and the extracellular space. The model is an extension of established cardiac bidomain models, which include a description of intra-myocyte and extracellular conductivities, currents and potentials in addition to transmembrane voltages of myocytes. Our extension added a description of fibroblasts, which are electrically coupled with each other and with myocytes. We applied the extended model in exemplary computational simulations of plane waves and conduction in a thin tissue slice assuming an isotropic conductivity of the intra-fibroblast domain. In simulations of plane waves, increased myocyte fibroblast coupling and fibroblast-myocyte ratio reduced peak voltage and maximal upstroke velocity of myocytes as well as amplitudes and maximal downstroke velocity of extracellular potentials. Simulations with the thin tissue slice showed that inter-fibroblast coupling affected rather transversal than longitudinal conduction velocity. Our results suggest that fibroblast coupling becomes relevant for small intra-myocyte and/or large intra-fibroblast conductivity. In summary, the study demonstrated the feasibility of the extended bidomain model and supports the hypothesis that fibroblasts contribute to cardiac electrophysiology in various manners. PMID- 19283481 TI - "She gives me a break from the world": formal youth mentoring relationships between adolescent girls and adult women. AB - Formal mentoring programs have historically tended to match youth with same-sex mentors; more recently, mentoring programs designed specifically for girls have begun cropping up in response to theories on gender and adolescent girls' psychological health and development, which suggest girls have particular psychosocial needs and ways of relating. Yet, there have been few empirical studies that explicitly examine the relational processes in adolescent girls' relationships with female mentors from the perspectives of the participants themselves. In the present study, qualitative interviews conducted with 12 female youth-adult pairs of participants (N = 24) in a one-to-one community-based mentoring program were analyzed thematically using a holistic-content approach. Examining these participants' narratives about their experiences in the program, three interrelated relational processes were identified: (a) engaged and authentic emotional support; (b) the development of new skills and confidence through collaborations; and, (c) experiences of companionship that provided relief from daily stresses. Editors' Strategic Implications: the focus on female dyads and relationships will richly inform further studies of the process of mentoring and provide insights for practitioners of a variety of gender-specific prevention programs. PMID- 19283482 TI - The short-term effectiveness of a suicide prevention gatekeeper training program in a college setting with residence life advisers. AB - Although the college years prove to be a vulnerable time for students and a critical period for suicide prevention, few school-based prevention strategies have been empirically evaluated. The current study examined the short-term effects of Question, Persuade, and Refer (QPR), a gatekeeper training program that teaches how to recognize warning signs, question suicidal intent, listen to problems, and refer for help. One hundred twenty-two residence advisers (RAs) who were trained in QPR demonstrated significant post-training gains across a variety of domains relevant to suicide and suicide prevention, with 60 who completed the follow-up assessment showing sustained knowledge and appraisals into the following semester. Although these gains were generally more substantial for RAs trained in QPR, 86 controls who completed both baseline and follow-up assessments also demonstrated changes in appraisals relevant to suicide and suicide prevention, despite having not received QPR training. Editors' Strategic Implications this study provides some promising evidence on gatekeeper training for RAs in terms of suicide prevention knowledge and attitudes. As the authors note, replication, rigorous evaluation, and studies of systemic efforts and behavioral outcomes are needed. PMID- 19283483 TI - Predictors of enrollment and retention in a preventive parenting intervention for divorced families. AB - Participation rates in parenting programs are typically low, severely limiting the public health significance of these interventions. We examined predictors of parenting program enrollment and retention in a sample of 325 divorced mothers. Predictors included intervention timing and maternal reports of child, parent, family, and sociocultural risk factors. In multivariate analyses, child maladjustment and family income-to-needs positively predicted enrollment, and higher maternal education and recruitment near the time of the divorce predicted retention. Findings have implications for the optimal timing of preventive parenting programs for divorcing families and point to the importance of examining predictors of enrollment and retention simultaneously. Editors' Strategic Implications: parent education researchers and practitioners may find the authors' application of the Health Belief Model to be a useful organizing framework for improving engagement and retention. PMID- 19283484 TI - Key theoretical frameworks for intervention: understanding and promoting behavior change in parent-infant feeding choices in a low-income population. AB - The early introduction of solids to infants is a risk factor associated with later health problems including allergies, overweight, and diabetes. The Infant Feeding Series (TIFS), a newly designed curriculum that promotes the appropriate transition to solids through parenting education and behavior change among low income mothers, used the Theory of Planned Behavior and the Transtheoretical Model of Change to develop TIFS curricular foci and activities. Using a pre-post design, pilot study results indicate that after exposure to the TIFS curriculum, mothers had significantly increased knowledge about appropriate infant feeding, could more accurately identify developmental indicators of infants' readiness for solids, and reported greater feelings of self efficacy about initiating and maintaining healthy feeding practices. Editors' Strategic Implications: replication is necessary, but TIFS appears to be a promising prevention program based on short-term knowledge and long-term behavioral outcomes (i.e., healthy feeding practices). PMID- 19283485 TI - Predicting participation in group parenting education in an Australian sample: the role of attitudes, norms, and control factors. AB - We examined the theory of planned behavior (TPB) in predicting intentions to participate in group parenting education. One hundred and seventy-six parents (138 mothers and 38 fathers) with a child under 12 years completed TPB items assessing attitude, subjective norms, perceived behavioral control (PBC), and two additional social influence variables (self-identity and group norm). Regression analyses supported the TPB predictors of participation intentions with self identity and group norm also significantly predicting intentions. Editors' Strategic Implications: these findings provide preliminary support for the TPB, in conjunction with additional sources of social influence, as a predictive model for participation in parent education and other prevention programs. PMID- 19283486 TI - Religion, health, and psychological well-being. AB - This study compares the effects of religiosity on health and well-being, controlling for work and family. With 2006 GSS data, we assess the effects of religiosity on health and well-being, net of job satisfaction, marital happiness, and financial status. The results indicate that people who identify as religious tend to report better health and happiness, regardless of religious affiliation, religious activities, work and family, social support, or financial status. People with liberal religious beliefs tend to be healthier but less happy than people with fundamentalist beliefs. Future research should probe how religious identity and beliefs impact health and well-being. PMID- 19283487 TI - Caspase-3 binds diverse P4 residues in peptides as revealed by crystallography and structural modeling. AB - Caspase-3 recognition of various P4 residues in its numerous protein substrates was investigated by crystallography, kinetics, and calculations on model complexes. Asp is the most frequent P4 residue in peptide substrates, although a wide variety of P4 residues are found in the cellular proteins cleaved by caspase 3. The binding of peptidic inhibitors with hydrophobic P4 residues, or no P4 residue, is illustrated by crystal structures of caspase-3 complexes with Ac-IEPD Cho, Ac-WEHD-Cho, Ac-YVAD-Cho, and Boc-D(OMe)-Fmk at resolutions of 1.9-2.6 A. The P4 residues formed favorable hydrophobic interactions in two separate hydrophobic regions of the binding site. The side chains of P4 Ile and Tyr form hydrophobic interactions with caspase-3 residues Trp206 and Trp214 within a non polar pocket of the S4 subsite, while P4 Trp interacts with Phe250 and Phe252 that can also form the S5 subsite. These interactions of hydrophobic P4 residues are distinct from those for polar P4 Asp, which indicates the adaptability of caspase-3 for binding diverse P4 residues. The predicted trends in peptide binding from molecular models had high correlation with experimental values for peptide inhibitors. Analysis of structural models for the binding of 20 different amino acids at P4 in the aldehyde peptide Ac-XEVD-Cho suggested that the majority of hydrophilic P4 residues interact with Phe250, while hydrophobic residues interact with Trp206, Phe250, and Trp214. Overall, the S4 pocket of caspase-3 exhibits flexible adaptation for different residues and the new structures and models, especially for hydrophobic P4 residues, will be helpful for the design of caspase-3 based drugs. PMID- 19283488 TI - Furano-sesquiterpene from soft coral, Sinularia kavarittiensis: induces apoptosis via the mitochondrial-mediated caspase-dependent pathway in THP-1, leukemia cell line. AB - Bioassay directed fractionation and purification led to the successful isolation of a furano sesquiterpene, Methyl 5-[(1E,5E)-2,6-Dimethyl octa-1,5,7-trienyl] furan-3-carboxylate (MDTFC), a bioactive component from a soft coral, Sinularia kavarittiensis. Its structure was determined by analyzing (1)H, (13)C NMR and FAB MS. The results show that MDTFC could efficiently and selectively inhibit the proliferation of several human cancer cell lines. Among all the cell lines, THP-1 was found to be most sensitive (IC(50) 29.59 microM), whereas the peripheral blood mononuclear cells were least effected (IC(50) 464.16 microM). The molecular mechanism of MDTFC mediated apoptosis was investigated for the first time. Induction of apoptosis in THP-1 cells was characterized by cell membrane blebbing, chromatin condensation, DNA fragmentation, and decrease in level of pro caspases 3, 9 and increase in Bax/Bcl-2 ratio. Our results were further strengthened through cleavage of poly (ADP-ribose) polymerase, reduction of mitochondrial membrane potential (Psim) and cytosolic release of cytochrome c, which are key events during apoptosis. Moreover, phosphatidyl serine exposure and appearance of sub-G1 peak also demonstrated cell death, when analyzed by flow cytometry. DNA fragmentation was prevented moderately when pretreated with caspase-9 inhibitor (Z-LEHD-FMK) and largely with caspase-3 inhibitor (Z-DEVD FMK). In summary, MDTFC mediated apoptosis involves mitochondria-dependent pathway and the present compound of marine origin might have a therapeutic value against human cancer cell lines and especially on leukemia cells. PMID- 19283489 TI - Health-related quality of life, quality-adjusted life years, and quality-adjusted life expectancy in new york city from 1995 to 2006. AB - We applied our previously developed estimation equation to predict EQ-5D index scores from the Centers for Disease Control and Prevention's Healthy Days measures for the New York City (NYC) adult population from 1995 to 2006 and compared these trends over time with the US general population. Such scores enabled us to examine the burden of disease attributable to smoking and overweight/obesity at both the local and national levels. We employed the estimation equation to the 1993-2007 Behavioral Risk Factor Surveillance System (BRFSS) data to obtain EQ-5D index scores for all survey respondents based on their age, self-rated health status, and overall number of unhealthy days. With the combination of mortality data, we calculated trends of quality-adjusted life years (QALYs), life expectancy (LE), and quality-adjusted life expectancy (QALE) as well as the percent of QALYs and QALE lost contributed by smoking and overweight/obesity. Mean EQ-5D index scores for NYC adults decreased from 0.874 to 0.852 but, more recently, have increased to 0.869. The LE of an 18-year-old living in NYC increased 4.7 years and QALE increased 2.6 years. The contribution of smoking to the proportion of QALYs lost decreased from 6.7% to 3.5%, while the contribution of overweight/obesity to the proportion of QALYs lost increased from 4.5% to 16.9%. The proportion of QALEs lost due to smoking decreased from 5.5% to 4.5%, while the proportion of QALEs lost due to overweight/obesity increased from 3.5% to 11.8%. Because the Healthy Days measures have been included in the BRFSS since 1993, translating Healthy Days Measures to a preference-based measure is a useful method for longitudinal tracking of population health at the local, state, and national level. PMID- 19283490 TI - Rhesus macaque model of chronic opiate dependence and neuro-AIDS: longitudinal assessment of auditory brainstem responses and visual evoked potentials. AB - Our work characterizes the effects of opiate (morphine) dependence on auditory brainstem and visual evoked responses in a rhesus macaque model of neuro-AIDS utilizing a chronic continuous drug delivery paradigm. The goal of this study was to clarify whether morphine is protective, or if it exacerbates simian immunodeficiency virus (SIV)-related systemic and neurological disease. Our model employs a macrophage tropic CD4/CCR5 coreceptor virus, SIV(mac)239 (R71/E17), which crosses the blood-brain barrier shortly after inoculation and closely mimics the natural disease course of human immunodeficiency virus infection. The cohort was divided into three groups: morphine only, SIV only, and SIV + morphine. Evoked potential (EP) abnormalities in subclinically infected macaques were evident as early as 8 weeks postinoculation. Prolongations in EP latencies were observed in SIV-infected macaques across all modalities. Animals with the highest cerebrospinal fluid viral loads and clinical disease showed more abnormalities than those with subclinical disease, confirming our previous work (Raymond et al., J Neurovirol 4:512-520, 1998; J Neurovirol 5:217-231, 1999; AIDS Res Hum Retroviruses 16:1163-1173, 2000). Although some differences were observed in auditory and visual evoked potentials in morphine-treated compared to morphine untreated SIV-infected animals, the effects were relatively small and not consistent across evoked potential type. However, morphine-treated animals with subclinical disease had a clear tendency toward higher virus loads in peripheral and central nervous system tissues (Marcario et al., J Neuroimmune Pharmacol 3:12 25, 2008) suggesting that if had been possible to follow all animals to end-stage disease, a clearer pattern of evoked potential abnormality might have emerged. PMID- 19283491 TI - Quality health care for cancer survivors: a survivor's perspective. PMID- 19283492 TI - Preformulation study of the inclusion complex irbesartan-beta-cyclodextrin. AB - The aim of the present work was to improve the solubility and dissolution profile of irbesartan (IRB), a poorly water-soluble drug by formation of inclusion complex with beta-cyclodextrin (betaCD). Phase solubility studies revealed increase in solubility of the drug upon cyclodextrin addition, showing A(L)-type of graph with slope less than one indicating formation of 1:1 stoichiometry inclusion complex. The stability constant (K(s)) was found to be 104.39 M(-1). IRB-betaCD binary systems were prepared by cogrinding, kneading using alcohol, kneading using aqueous alcohol, and coevaporation methods. Characterization of the binary systems were carried out by differential scanning calorimetry, Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction, and proton nuclear magnetic resonance. The dissolution profiles of inclusion complexes were determined and compared with those of IRB alone and physical mixture. Among the various methods, coevaporation was the best in which the solubility was increased and dissolution rate of the drug was the highest. The study indicated the usefulness of cyclodextrin technology to overcome the solubility problem of IRB. PMID- 19283493 TI - Immobilising of Cd, Pb, and Zn contaminated arable soils close to a former Pb/Zn smelter: a field study in Austria over 5 years. AB - Numerous smelter sites are surrounded by rural land. The entrance of non essential metals such as lead or cadmium into the food chain is very likely as well as phytotoxicity effects of zinc. Finding a realistic solution for these large-scale contaminations was one aim of this study. Previous results from pot experiments showed a high potential for the reduction of metals entering the food chain via crops grown on smelter-contaminated soils from Arnoldstein, Austria, by the use of amendments for immobilisation. A further aim was to optimise a field experiment for overcoming the gap between pot and field experiments and to look for long-term efficiency of the treatments [lime (CA), red mud (RM), gravel sludge + red mud (GS + RM)]. Field experiment results were obtained for 5 years. Besides soil and soil pore water samples, the following harvests were yielded: spring barley (Hordeum distichon ssp. L.) (2004-2005), narrowleaf plantain (Plantago lanceolata L.) (2006-2007) and velvet grass (Holcus lanatus L.) (2007 2008). The long-term efficiency of GS + RM led us to conclude that their application seems to be a realistic and practical measure for extensively contaminated land, best in combination with metal excluding cultivars. PMID- 19283494 TI - Relationships of self-reported physical activity domains with accelerometry recordings in French adults. AB - The objective was to examine the relationships of self-reported physical activity (PA) by domain (leisure, occupational, other) with PA and sedentary time as measured objectively by accelerometry. Subjects were adults with low habitual PA levels from a community in northern France. Among subjects in the lowest tertile of a PA score from a screening questionnaire, 160 (37% males, age: 41.0 +/- 10.8 years, BMI: 25.1 +/- 4.1 kg/m(2), mean +/- SD) completed a detailed instrument (Modifiable Activity Questionnaire), and wore an accelerometer (Actigraph) for seven consecutive days. Relationships between questionnaire domains (occupational, leisure, and "non-occupational non-leisure") and accelerometry measures (total activity and sedentary time) were assessed using Spearman correlation coefficients. In this population, the highest contributor to total reported PA (h/week) was occupational PA. Time spent in non-occupational non leisure PA ranked second in women and third in men. The most frequent non occupational non-leisure PA were shopping and household chores. In women, non occupational non-leisure PA contributed more than occupational or leisure-time PA to total PA energy expenditure (median: 18.0, 9.1, and 4.9 MET-h/week, respectively). Total PA by accelerometry (count/day) was correlated to leisure time PA in women (r = 0.22, P < 0.05) and to occupational (r = 0.43, P < 0.01) and total reported PA (r = 0.39, P < 0.01) in men (all in MET-h/week). There was an inverse relationship between accelerometry sedentary time (h/day) and non occupational non-leisure PA (MET-h/week, r = -0.30, P < 0.001). These findings indicate the importance of assessing non-occupational non-leisure PA for a better understanding of how individuals partition their time between active or sedentary occupations. PMID- 19283495 TI - Chromosomal rearrangements underlying karyotype differences between Chinese pangolin (Manis pentadactyla) and Malayan pangolin (Manis javanica) revealed by chromosome painting. AB - The Chinese pangolin (Manis pentadactyla), a representative species of the order Pholidota, has been enlisted in the mammalian whole-genome sequencing project mainly because of its phylogenetic importance. Previous studies showed that the diploid number of M. pentadactyla could vary from 2n = 36 to 42. To further characterize the genome organization of M. pentadactyla and to elucidate chromosomal mechanism underlying the karyotype diversity of Pholidota, we flow sorted the chromosomes of 2n = 40 M. pentadactyla, and generated a set of chromosome-specific probes by DOP-PCR amplification of flow-sorted chromosomes. A comparative chromosome map between M. pentadactyla and the Malayan pangolin (Manis javanica, 2n = 38), as well as between human and M. pentadactyla, was established by chromosome painting for the first time. Our results demonstrate that seven Robertsonian rearrangements, together with considerable variations in the quantity of heterochromatin and in the number of nucleolar organizer regions (NORs) differentiate the karyotypes of 2n = 38 M. javanica and 2n = 40 M. pentadactyla. Moreover, we confirm that the M. javanica Y chromosome bears one NOR. Comparison of human homologous segment associations found in the genomes of M. javanica and M. pentadactyla revealed seven shared associations (HSA 1q/11, 2p/5, 2q/10q, 4p+q/20, 5/13, 6/19p and 8q/10p) that could constitute the potential Pholidota-specific signature rearrangements. PMID- 19283496 TI - A plot of G + C content against sequence length of 640 bacterial chromosomes shows the points are widely scattered in the upper triangular area. AB - In this work, we study the distribution of G + C content against chromosome size among 640 fully sequenced bacterial chromosomes. When %G + C is plotted versus increasing chromosome length, it is shown that the points denoting 640 bacterial chromosomes are widely scattered in the upper triangular area and the triangle is right-angled. The mean G + C content increases while the range or variance of G + C contents decreases with length. The net effect of these two trends is to exclude data from a triangular area that should represent long A + T-rich chromosomes. Thus, the shorter chromosomes have a wide, 20-70%, range of G + C contents while the longest chromosomes are severely restricted to a G + C content near 70%. To generate the distribution type of right-angled triangle, two conditions are requisite. Consequently, two possible related explanations are presented to interpret the relationship. One aims to give the reason why most large bacterial chromosomes have high G + C contents and the other tries to interpret why most A + T-rich bacterial chromosomes tend to be small in size. Both of the two interpretations are proposed from the selectionist viewpoint and the latter is an addition to the former one. According to the theories, the bacteria with large chromosomes and bacteria with A + T-rich chromosomes have different tactics to cope with ultraviolet radiation and other physical or chemical damage. PMID- 19283497 TI - Organization of the amplified type I interferon gene cluster and associated chromosome regions in the interphase nucleus of human osteosarcoma cells. AB - The organization of the amplified type I interferon (IFN) gene cluster and surrounding chromosomal regions was studied in the interphase cell nucleus of the human osteosarcoma cell line MG63. Rather than being arranged in a linear ladder like array as in mitotic chromosomes, a cluster of approximately 15 foci was detected that was preferentially associated along the periphery of both the cell nucleus and a chromosome territory containing components of chromosomes 4, 8, and 9. Interspersed within the IFN gene foci were corresponding foci derived from amplified centromere 4 and 9 sequences. Other copies of chromosomes 4 and 8 were frequently detected in pairs or higher-order arrays lacking discrete borders between the chromosomes. In contrast, while chromosomes 4 and 8 in normal WI38 human fibroblast and osteoblast cells were occasionally found to associate closely, discrete boundaries were always detected between the two. DNA replication timing of the IFN gene cluster in early- to mid-S phase of WI38 cells was conserved in the amplified IFN gene cluster of MG63. Quantitative RT-PCR demonstrated a approximately 3-fold increase in IFN beta transcripts in MG63 compared with WI38 and RNA/DNA FISH experiments revealed 1-5 foci of IFN beta transcripts per cell with only approximately 5% of the cells showing foci within the highly amplified IFN gene cluster. PMID- 19283498 TI - Mercury concentrations in fish from a Sierra Nevada foothill reservoir located downstream from historic gold-mining operations. AB - This study examined mercury concentrations in whole fish from Camp Far West Reservoir, an 830-ha reservoir in northern California, USA, located downstream from lands mined for gold during and following the Gold Rush of 1848-1864. Total mercury (reported as dry weight concentrations) was highest in spotted bass (mean, 0.93 microg/g; range, 0.16-4.41 microg/g) and lower in bluegill (mean, 0.45 microg/g; range, 0.22-1.96 microg/g) and threadfin shad (0.44 microg/g; range, 0.21-1.34 microg/g). Spatial patterns for mercury in fish indicated high concentrations upstream in the Bear River arm and generally lower concentrations elsewhere, including downstream near the dam. These findings coincided with patterns exhibited by methylmercury in water and sediment, and suggested that mercury-laden inflows from the Bear River were largely responsible for contaminating the reservoir ecosystem. Maximum concentrations of mercury in all three fish species, but especially bass, were high enough to warrant concern about toxic effects in fish and consumers of fish. PMID- 19283499 TI - Antioxidative responses related to H(2)O(2) depletion in Hypnum plumaeforme under the combined stress induced by Pb and Ni. AB - The short-term responses of H(2)O(2)-depletion-related parameters in moss Hypnum plumaeforme to the combined stress induced by Pb and Ni were investigated. The results showed that the Pb and Ni stress induced dose-dependent accumulation of hydrogen peroxide (H(2)O(2)). The increase of peroxidase (POD) activity and decrease of ascorbate peroxidase (APX) activity were observed under the combined heavy metal application. The antioxidants, ascorbate (AsA) and proline content, increased significantly when the metals were applied together. The study indicated that the cell damage caused by Pb stress was higher than that caused by Ni stress, Pb and Ni had synergistic effect in inducing the oxidative stress in moss H. plumaeforme, especially under the combination of high concentration of Ni (0.1 and 1.0 mM) and Pb. Content of proline, H(2)O(2) and the activity of POD, all showed a dose-dependent increase under Pb and Ni stress, suggesting their practical value as biomarkers in moss biomonitoring, especially in the case of light pollution caused by heavy metals without the changes in the appearance of mosses. PMID- 19283500 TI - Correlation of United States Medical Licensing Examination and Internal Medicine In-Training Examination performance. AB - The Internal Medicine In-Training Examination (ITE) is administered during residency training in the United States as a self-assessment and program assessment tool. Performance on this exam correlates with outcome on the American Board of Internal Medicine Certifying examination. Internal Medicine Program Directors use the United States Medical Licensing Examination (USMLE) to make decisions in recruitment of potential applicants. This study was done to determine a correlation of USMLE Steps 1, 2 and 3 results with ITE scores in each level of Internal Medicine training. A retrospective review of all residents graduating from an Internal Medicine program from 1999 to 2006 was done. Subjects included had data for all USMLE Steps and ITE during all years of training. Thirty-one subjects were included in the study. Correlations of USMLE Steps 1, 2 and 3 were done with ITE scores (percent correct) in each year of training. Pearson's correlation coefficient (r) was determined for each pairing and a t test to determine statistical significance of the correlation was done. Statistical significance was defined as P value <0.05. The r values for USMLE Step 1 and ITE percent correct in PGY I, II and III were 0.46, 0.55 and 0.51 respectively. Corresponding r values for USMLE Step 2 and ITE percent correct were 0.79, 0.70 and 0.72; for USMLE Step 3 these values were 0.51, 0.37 and 0.51 respectively for each training year. USMLE scores are correlated with ITE scores. This correlation was strongest for USMLE Step 2. PMID- 19283501 TI - Synthesis and palladium-catalysed isomerisation of fused polycyclic tetrahydrofurans: efficient and stereoselective one-pot domino construction of functionalised bridged bicyclo[n.2.1] ring systems. AB - A new one-pot domino reaction for a general entry to functionalised bridged bicyclo[n.2.1] ring systems from alpha, alpha'-diactivated cyclic ketones and trans-1,4-dihalides is described. The sequence combines a base promoted C-O cycloalkylation reaction leading to fused polycyclic enol ethers and their in situ palladium-catalysed isomerisation. PMID- 19283502 TI - A reinvestigation of the acid-promoted heterocyclization of 2-(2-oxo-2 arylethyl)malononitriles in the presence of amines. AB - The reinvestigation of the acid-promoted cyclization of 2-(2-oxo-2 arylethyl)malononitriles, in the presence of benzylamine or aniline, in ethanol or acetonitrile, has confirmed that this is a long-time reaction process for a low-yielding synthesis of 2-amino-5-arylfuran-3-carbonitriles (2), or 2-amino-5 aryl-1-phenyl-1H-pyrrole-3-carbonitriles (4), depending on the base used. However, the microwave-assisted synthesis of 2-amino-5-(4'-methoxyphenyl)furan 3(4)-(di)carbonitriles (2c and 3c) proceeds in shorter reaction times and higher yields than does the classical thermal heating protocol. In these reactions we have observed for the first time, and characterized by their spectroscopic data and X-ray analysis, the unexpected formation of 2-amino-5-aryl-3 (4) (di)carbonitriles (3), whose formation has been rationalized by density functional theory (DFT) analysis of the proposed reaction mechanism. PMID- 19283503 TI - Diastereoselective synthesis of fused [1,3]thiazolo[1,3]oxazins and [1,3]oxazino[2,3-b][1,3]benzothiazoles. AB - An efficient diastereoselective synthesis of 7-ethyl 5,6-dialkyl 7H [1,3]thiazolo[2,3-b][1,3]oxazin-5,6, 7-tricarboxylates and 2-ethyl 3,4-dialkyl 2H [1,3]oxazino [2,3-b][1,3]benzothiazole-2,3,4-tricarboxylates via reaction of thiazole and benzothiazole with dialkyl acetylenedicarboxylates in the presence of ethyl pyruvate is described. PMID- 19283504 TI - Bacterial fermentation in the gastrointestinal tract of non-ruminants: influence of fermented feeds and fermentable carbohydrates. AB - The search for alternatives to in-feed antibiotics in animal nutrition has highlighted the role dietary modulation can play in improving gut health. Current antibiotic replacement strategies have involved the use of microbes beneficial to health (probiotics) or fermentable carbohydrates (prebiotics) or both (synbiotics). The present review recognises the contribution of fermented feeds and fermentable carbohydrates in improving the gut environment in non-ruminants. It proposes the screening of probiotic bacteria for the production of fermented feeds and supplementation of these feeds with fermentable carbohydrates prior to feeding animals. It is suggested that the term 'fermbiotics' should be used to describe this intervention strategy. PMID- 19283505 TI - Body mass index modulates postural proteinuria. PMID- 19283506 TI - CD44 expression in renal ischemia-reperfusion injury in rats. AB - Renal ischemia-reperfusion injury (IRI) is an invariable consequence of transplantation. The tubuloepithelial expression of CD44 is markedly enhanced in autoimmune renal injuries. The aim of this experimental study was to evaluate the effect of IRI on the expression of CD44 in rat kidney. Thirty male Sprague-Dawley rats were used. The rats were divided into three groups. The rats in group 1 (n = 10) underwent laparotomy and left nephrectomy (Sham surgery). The rats in group 2 (n = 10) underwent laparotomy, 1 h renal ischemia, followed by 1 h of reperfusion and then left nephrectomy was performed. The rats in group 3 (n = 10) underwent laparotomy, 1 h renal ischemia, followed by 24 h of reperfusion and then left nephrectomy was performed. Histopathological findings and the immunohistochemical expression of CD44 in ischemic and reperfused rat kidneys were investigated. In histopathologic evaluation, non-specific changes were observed in group 2 and early phase of IRI were present in group 3. CD44 was expressed in both group 2 and 3 but not in group 1. The mean immunohistochemical staining percentages of rat kidneys in group 1, 2, and 3 were 0.00 +/- 0.00, 39.90 +/- 5.53, and 26.20 +/ 8.38, respectively. The immunohistochemical staining pattern was more dense in group 2 than in group 3 (P < 0.001). In conclusion, the expression of CD44 in renal tubuloepithelial cells was significantly increased after IRI. The increase in CD44 expression was more prominent during the early phase of IRI and started to decline after 24 h of reperfusion. PMID- 19283507 TI - Clinical symptoms related to renal cell carcinoma are independent prognostic factors for intraoperative complications and overall survival. AB - OBJECTIVE: It is not yet elucidated whether the symptoms related to renal cell carcinoma have a strong effect on intraoperative complications or survival. We aimed to investigate this association in a cohort of renal cancer patients operated on between June 1997 and December 2004 at the Department of Urology, Semmelweis University School of Medicine. PATIENTS AND METHODS: Among 363 consecutive patients with renal masses treated at our institution, only 200 (55.3%) were truly asymptomatic and completely incidental (group A). Among the 259 patients with incidentally detected tumors, 59 (16.1%) had symptoms that were probably related to the renal lesion according to a reviewed history (group B) and 104 patients (28.6%) presented for symptoms related to renal cell cancer (group C). RESULTS: Patients in group B had a higher risk for weight loss (P < 0.0001) and flank pain (P = 0.063), lower risk for symptoms related to distant metastases, while their outcome was not significantly different from group C. The collapsed group (B + C) had an increased risk for metastasis (P = 0.002), higher stage (P = 0.001), and intraoperative complications (P = 0.046) compared to group A according to a logistic regression. The presence of symptoms was significantly related to the overall survival using the Kaplan-Meier method (P < 0.0001). CONCLUSIONS: Weight loss and flank pain were frequently neglected as a possible symptom of renal cancer by the patients. An easily accessible parameter (the presence of symptoms) was independently related to intraoperative complications and survival in renal cancer patients. PMID- 19283508 TI - Growth pattern in superficial urothelial bladder carcinomas. Histological review and clinical relevance. AB - INTRODUCTION: The question of when an intraepithelial urothelial carcinoma becomes invasive into the lamina propria of the urinary bladder is an unresolved issue. Our objective was to analyse a series of consecutive superficial carcinomas to assess the importance of growth pattern in tumour recurrence and progression. MATERIALS AND METHODS: The pathological staging of 200 superficial (pTa/pT1) bladder carcinomas was reviewed. Non-invasive lesions and tumours invading the lamina propria were distinguished. Both infiltrating and pushing patterns of growth were regarded as lamina propria invasion. RESULTS: A total of 35 (17.5%) pTa and 165 (82.5%) pT1 tumours were identified. Among pT1 tumours, 39 (23.6%) displayed the infiltrating pattern of invasion and 126 (76.4%) the pushing pattern. Differences in five-year recurrence-free (P = 0.01) and progression-free (P = 0.001) survival were demonstrated between pTa and pT1 tumours, and between pT1 infiltrating and pT1 pushing subcategories. Invasive growth pattern has a 1.86 times higher risk of tumour recurrence and 3.01 times higher risk of progression. CONCLUSIONS: The pT1 category of bladder carcinoma should include a group of tumours defined by its pushing pattern of growth. Some cases may have been previously considered pTa, but follow an intermediate clinical course between pTa and pT1 tumours with infiltrating growth pattern. PMID- 19283509 TI - Measurement of D: -2-hydroxyglutarate dehydrogenase activity in cell homogenates derived from D: -2-hydroxyglutaric aciduria patients. AB - D: -2-Hydroxyglutaric aciduria (D: -2-HGA) is a neurometabolic disorder characterized by elevated levels of D: -2-hydroxyglutarate (D: -2-HG) in physiological fluids. Recent findings revealed that mutations in the D2HGDH gene, encoding D: -2-hydroxyglutarate dehydrogenase, cause D: -2-HGA. So far, a functionalenzyme assay to determine D: -2-hydroxyglutarate dehydrogenase activity, converting D: -2-HG into 2-ketoglutarate (2-KG), has been unavailable. We have now developed a unique enzyme assay for the determination of D: -2 hydroxyglutarate dehydrogenase activity in cells derived from D: -2-HGA patients and controls. The enzyme assay was performed using enantiomerically pure stable isotope-labelled D: -2-hydroxy[3,3,4,4-(2)H(4)]glutarate. This substrate is convertedby D: -2-hydroxyglutarate dehydrogenase into 2-[3,3,4,4 (2)H(4)]ketoglutarate, which is subsequently converted into L: -[3,3,4,4 (2)H(4)]glutamate by L: -glutamate dehydrogenase, present in saturating amounts in cell homogenates. Enzyme activities were quantified using LC-MS/MS. The mean activities in control fibroblast and lymphoblast homogenates were 298 +/- 207 and 1670 +/- 940 pmol/h per mg protein, respectively. In fibroblast and lymphoblast cell lines derived from patients with pathogenic mutations in the D2HGDH gene, considerably decreased enzyme activities (e.g. <41 pmol/h per mg protein) were found compared with controls. This enzyme assay will have additional utility in further differentiating patients with D: -2-HGA and L: -2-HGA and in assessing the residual activities linked to pathogenic mutations in the D2HGDH gene. PMID- 19283510 TI - Polymorphism Val103Ile of the melanocortin-4 receptor gene in the Serbian population. AB - The melanocortin-4 receptor (MC4R) plays an important role in weight and energy homeostasis and it is associated with lower risk to develop obesity and lower body mass index. The contribution of MC4R mutation to obesity in Vojvodina (Northern Province of Serbia), known as a region with the largest number of overweight people, has not been previously investigated. The objective of this study was to examine the Val103Ile polymorphism of MC4R in a population of Vojvodina and its association with obesity. The study was carried out in a group of 96 persons: 62 obese and 34 normal weight men and women. Anthropometric measurements and cardiovascular risk factors assessment were done. The genotypes were determined by PCR-RFLP. In our on going study, three subjects were heterozygous for Val103Ile mutation (3.12%), and one was homozygous for 103Ile allele (1.04%). Among obese patients no isoleucine allele homozygous was found. The frequencies of the 103Ile allele in a group of obese and normal weight persons were found to be 1.61 and 4.41%, respectively. Val103Ile polymorphism of melanocortin-4 receptor is unlikely to be a major cause of overweight and obesity in Vojvodina, but further studies on larger groups of patients are needed. PMID- 19283511 TI - A coding SNP of LHX4 gene is associated with body weight and body length in bovine. AB - Heterozygous mutations in LHX4 are associated with combined pituitary hormone deficiency. In this study, the polymorphism of LHX4-HaeIII locus was revealed in 822 individuals from four Chinese cattle breeds. The PCR-RFLP analysis showed that there were three genotypes: GG, GA, AA. The frequencies of genotype GG ranged from 0.6620 to 0.9789 in analyzed populations. The genotypic frequencies of LHX4 locus in the four populations all agreed with Hardy-Weinberg equilibrium (P > 0.05). Distributions of genotypic frequencies of different breeds (QC, NY, JX, CH) at this locus were found to be significantly different based on a chi(2) test (P < 0.001). The genetic diversity analysis revealed the JX cattle possessed intermediate genetic diversity, and the other three Chinese cattle breeds belonged to poor genetic diversity. Correlation analysis with growth traits in the NY breed indicated that: the animals with genotype GA had greater body weight than those with genotype GG (P < 0.05); the animals with GA genotype owned significantly longer body length than the ones with GG genotype (P < 0.05) at 18 and 24 months. PMID- 19283513 TI - A theory of health science and the healing arts based on the philosophy of Bernard Lonergan. AB - This paper represents a preliminary investigation relating Bernard Lonergan's thought to health science and the healing arts. First, I provide background for basic elements of Lonergan's theoretical terminology that I employ. As inquiry is the engine of Lonergan's method, next I specify two questions that underlie medical insights and define several terms, including health, disease, and illness, in relation to these questions. Then I expand the frame of reference to include all disciplines involved in the cycle of clinical interaction under the heading health science and the healing arts. Finally, I analyze the cycle of clinical interaction in terms of Lonergan's cognitive theory. I compare and contrast my analysis, based on Lonergan, with that of Pellegrino, Thomasma and Sulmasy as I proceed. In closing, I comment briefly on the next stage of this project regarding Lonergan's theory of the human good in relation to the practice of the healing arts. PMID- 19283512 TI - Functional polymorphisms of matrix metallopeptidase-9 and risk of coronary artery disease in a Chinese population. AB - Matrix metallopeptidase-9 (MMP-9) plays a pivotal role in vascular remodeling and development of atherosclerotic lesion. The potentially functional MMP-9 polymorphisms may contribute to the susceptibility of coronary artery disease (CAD). A case-control study composed of 762 CAD cases and 555 CAD-free controls was conducted in a Chinese population to investigate the association between the MMP-9 -1562 C>T, R279Q, P574R and R668Q polymorphisms and CAD risk. It was found that the variant genotypes of R279Q, P574R and R668Q were associated with a non significant decreased risk of CAD when compared with their wild-type genotypes, respectively, Furthermore, compared with those without any variant genotypes for these four nonsynonymouse loci, individuals carrying all four variant genotypes ( 1562 CT/TT, 279 RQ/QQ, 574 PR/RR and 668 RQ/QQ) had a 51% decreased risk of CAD (adjusted OR = 0.49; 95% CI = 0.26-0.95, P = 0.033). Although no significant main effects were observed for MMP-9 -1562 C>T locus on CAD risk, variant genotypes of -1562 C>T were associated with a 2.53 increased risk of CAD in subjects with diabetes mellitus (DM) (95% CI = 1.18-5.45, P = 0.018). In CAD cases, variant genotypes of -1562 C>T were associated with a significantly increased risk of MI (adjusted OR, 1.48, 95% CI, 1.01-2.20, P = 0.048). These findings suggest that MMP-9 R279Q, P574R and R668Q may have combined effect in the occurrence of CAD and -1562 CT/TT genotypes may contribute to CAD in diabetics and MI in CAD patients. PMID- 19283515 TI - High-level expression and novel antifungal activity of mouse beta defensin-1 mature peptide in Escherichia coli. AB - Mouse beta defensin-1 (mBD-1) is a cationic 37-amino acid antimicrobial peptide with three conserved cysterine disulfied bonds. It exhibits a broad antimicrobial spectrum, but mBD-1 against Candida albicans (C. albicans) and Cryptococcus neoformans (C. neoformans) is poorly understood. This study describes the mBD-1 gene, the heterologous fusion expression of the peptide in Escherichia coli, and the bioactive assay of released mature mBD-1. By constructing the expression plasmid (pET32a-mBD1), high yields of soluble mBD-1 fusion protein (0.67 g/L) could be obtained in E. coli and cleaved by enterokinase. The digested product was further purified and desalted with the final amount of pure mature mBD-1 being 0.14 g/L. Classical fungi growth inhibition assay showed clear antifungal activity against C. albicans and C. neoformans with IC(50) of 5 and 2 microM, respectively. The results show that the mBD-1 control fungal colonization through hyphal induction, direct fungicidal activity, and the activity is suppressed by increasing NaCl concentration. Successful expression of the mBD-1 peptide in E. coli offers a basis for further studying its antifungal mechanisms and may provide significance in developing this peptide to an antifungal drug. PMID- 19283517 TI - Bacterial attachment and removal properties of silicon- and nitrogen-doped diamond-like carbon coatings. AB - Si- and N-doped diamond-like carbon (DLC) coatings with various Si and N contents were deposited on glass slides using magnetron sputter ion-plating and plasma enhanced chemical vapour deposition. Surface energy analysis of the DLC coatings revealed that with increasing Si content, the electron acceptor gamma(s)(+) value decreased while the electron donor gamma(s)(-) value increased. The antifouling property of DLC coatings was evaluated with the bacterium, Pseudomonas fluorescens, which is one of the most common microorganisms forming biofilms on the surface of heat exchangers in cooling water systems. P. fluorescens had a high value of the gamma(s)(-) component (69.78 mN m(-1)) and a low value of the gamma(s)(+) component (5.97 mN m(-1)), and would be negatively charged with the zeta potential of -16.1 mV. The experimental results showed that bacterial removal by a standardised washing procedure increased significantly with increasing electron donor gamma(s)(-) values and with decreasing electron acceptor gamma(s)(+) values of DLC coatings. The incorporation of 2%N into the Si doped DLC coatings further significantly reduced bacterial attachment and significantly increased ease of removal. The best Si-N-doped DLC coatings reduced bacterial attachment by 58% and increased removal by 41%, compared with a silicone coating, Silastic T2. Bacterial adhesion strength on the DLC coatings is explained in terms of thermodynamic work of adhesion. PMID- 19283518 TI - Welcome to a new style biomarkers from the new editor in chief. PMID- 19283519 TI - Manganese exposure among smelting workers: blood manganese-iron ratio as a novel tool for manganese exposure assessment. AB - Unexposed control subjects (n = 106), power distributing and office workers (n = 122), and manganese (Mn)-exposed ferroalloy smelter workers (n = 95) were recruited to the control, low and high groups, respectively. Mn concentrations in saliva, plasma, erythrocytes, urine and hair were significantly higher in both exposure groups than in the controls. The Fe concentration in plasma and erythrocytes, however, was significantly lower in Mn-exposed workers than in controls. The airborne Mn levels were significantly associated with Mn/Fe ratio (MIR) of erythrocytes (eMIR) (r = 0.77, p < 0.01) and plasma (pMIR) (r = 0.70, p < 0.01). The results suggest that the MIR may serve as a useful biomarker to distinguish Mn-exposed workers from the unexposed, control population. PMID- 19283516 TI - Purinoceptors on neuroglia. AB - Purinergic transmission is one of the most ancient and widespread extracellular signalling systems. In the brain, purinergic signalling plays a unique role in integrating neuronal and glial cellular circuits, as virtually every type of glial cell possesses receptors to purines and pyrimidines. These receptors, represented by metabotropic P1 adenosine receptors, metabotropic P2Y purinoceptors and ionotropic P2X purinoceptors, control numerous physiological functions of glial cells and are intimately involved in virtually every form of neuropathology. In this essay, we provide an in depth overview of purinoceptor distribution in two types of CNS glia--in astrocytes and oligodendrocytes--and discuss their physiological and pathophysiological roles. PMID- 19283520 TI - Ethane and n-pentane in exhaled breath are biomarkers of exposure not effect. AB - The relationship of exhaled ethane and n-pentane to exhaled NO, carbonylated proteins, and indoor/outdoor atmospheric pollutants were examined in order to evaluate ethane and n-pentane as potential markers of airway inflammation and/or oxidative stress. Exhaled NO and carbonylated proteins were found to have no significant associations with either ethane (p = 0.96 and p = 0.81, respectively) or n-pentane (p = 0.44 and 0.28, respectively) when outliers were included. In the case where outliers were removed n-pentane was found to be inversely associated with carbonylated proteins. Exhaled hydrocarbons adjusted for indoor hydrocarbon concentrations were instead found to be positively associated with air pollutants (NO, NO(2) and CO), suggesting pollutant exposure is driving exhaled hydrocarbon concentrations. Given these findings, ethane and n-pentane do not appear to be markers of airway inflammation or oxidative stress. PMID- 19283521 TI - The acute-phase protein serum amyloid A3 is expressed in the bovine mammary gland and plays a role in host defence. AB - The serum amyloid A protein is one of the major reactants in the acute-phase response. Using representational difference analysis comparing RNA from normal and involuting quarters of a dairy cow mammary gland, we found an mRNA encoding the SAA3 protein (M-SAA3). The M-SAA3 mRNA was localized to restricted populations of bovine mammary epithelial cells (MECs). It was expressed at a moderate level in late pregnancy, at a low level through lactation, was induced early in milk stasis, and expressed at high levels in most MECs during mid to late involution and inflammation/mastitis. The mature M-SAA3 peptide was expressed in Escherichia coli, antibodies made, and shown to have antibacterial activity against E. coli, Streptococcus uberis and Pseudomonas aeruginosa. These results suggest that the mammary SAA3 may have a role in protection of the mammary gland during remodelling and infection and possibly in the neonate gastrointestinal tract. PMID- 19283522 TI - Increased ischaemia modified albumin following coronary artery bypass grafting. AB - BACKGROUND: Any increase of cardiac biomarkers after coronary artery bypass grafting (CABG) indicates myocyte necrosis and is likely to be related to an impaired outcome. We investigated whether ischaemia-modified albumin (IMA), a biomarker of ischaemia, is also raised following CABG. METHODS: We studied 50 stable consecutive patients undergoing elective isolated CABG on cardiopulmonary bypass, of whom 46 were men and four women, aged 64 +/- 9 years. Blood samples were obtained the day before the operation (pre-op) as well as immediately after the operation, 24 h postoperatively (post-op) and the fourth day post-op and assayed for creatine kinase, the MB isoenzyme of creatine kinase, cardiac troponin-I, albumin and IMA. RESULTS: The typical rising and falling pattern of myocardial necrosis of all three cardiac enzymes was observed post-op (p <0.0001). IMA increased significantly following CABG at all three time points (113 +/- 43, 106.7 +/- 22.6 and 110.2 +/- 12.5 U ml(-1), respectively) compared with pre-op values (91.7 +/- 10.5 U ml(-1)), (p <0.0001); the sample immediately post-op was significantly higher compared with the following samples (immediately post-op vs 24 h, p = 0.008 and immediately post-op vs 4 days, p = 0.03, with no significant difference between the last two). IMA level changes during the study course were independent of the albumin changes. Haemoglobin decreased significantly post-op (p <0.0001 vs baseline) whereas serum creatinine did not differ during the study period. CONCLUSIONS: IMA increases significantly following CABG but whether or not this carries a prognostic significance remains to be elucidated. PMID- 19283523 TI - Does ischemia-modified albumin add prognostic value to the Thrombolysis In Myocardial Infarction risk score in patients with ST-segment elevation myocardial infarction treated with primary angioplasty? AB - BACKGROUND: The aim of the present study was to evaluate whether or not an elevated ischaemia-modified albumin (IMA) level provides any additional prognostic information to the validated Thrombolysis In Myocardial Infarction (TIMI) risk score in patients with ST-segment elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI). METHODS: One hundred seven consecutive STEMI patients treated with primary PCI were included. The incidence of 30-day death was the prespecified primary end point. Serum IMA was measured immediately at hospital arrival. RESULTS: The incidence of the primary end point was 6.5%. A significant predictive value of IMA in relation to the primary end point was indicated by an area under the ROC curve of 0.71 (p = 0.01). In the multivariate analysis, increased IMA remained a significant predictor of the primary end point after adjustment for TIMI risk predictors (p = 0.019). The area under the ROC curve for the TIMI risk score was 0.68 (p = 0.03). The addition of IMA to the TIMI risk score did not improve its prognostic value (area under the ROC curve 0.60, p = 0.25). CONCLUSION: IMA levels obtained at admission are a powerful indicator of short-term mortality in STEMI patients treated with primary PCI, but do not seem to be a marker that adds prognostic information to the validated STEMI TIMI risk score. PMID- 19283524 TI - Plasma total homocysteine level and methylenetetrahydrofolate reductase 677C>T genetic polymorphism in Japanese patients with rheumatoid arthritis. AB - Hyperhomocysteinemia is a known risk factor of cardiovascular disease. Homocysteine has been also linked to inflammation in rheumatoid arthritis (RA). In this study, we investigated the relationship between plasma homocysteine levels and single nucleotide polymorphism (SNP) of the gene coding for methylenetetrahydrofolate reductase (MTHFR), an enzyme involved in the biosynthesis of homocysteine, and the correlation between the plasma homocysteine levels and generally used inflammatory markers (C-reactive protein, erythrocyte sedimentation rate and matrix metalloproteinase-3) in 96 Japanese patients with RA. Plasma homocysteine levels in patients with the MTHFR 677TT genotype were significantly higher than in those with the 677CC genotype (p < 0.05). In addition, plasma homocysteine levels were increased along with the elevation of general inflammatory markers. Therefore, we conclude that homocysteine might affect the inflammatory status of patients, and the MTHFR 677C>T SNP could be a predictive factor of hyperhomocysteinemia in patients with RA. PMID- 19283525 TI - Metabolomic analysis of bronchoalveolar lavage fluid from cystic fibrosis patients. AB - Metabolite profiles of bronchoalveolar lavage fluid (BALF) from pediatric patients with cystic fibrosis (CF) were correlated to the degree of airway inflammation using nuclear magnetic resonance spectroscopy-based metabolomics. BALF was collected from 11 children with CF during clinically indicated bronchoscopy. The spectra from BALF with high levels of neutrophilic airway inflammation displayed signals from numerous metabolites, whereas the spectra from subjects with low levels of inflammation were very sparse. The metabolites identified in samples taken from subjects with high inflammation include known markers of inflammation such as amino acids and lactate, as well as many novel signals. Statistical analysis highlighted the most important metabolites that distinguished the high- from the low-inflammation groups. This first demonstration of metabolomics of human BALF shows that clear distinctions in the metabolic profiles can be observed between subjects experiencing high versus low inflammation and is a first step toward the goal of discovering novel biomarkers of airway inflammation. PMID- 19283526 TI - Serum growth factors in asbestosis patients. AB - Various growth factors, including platelet-derived growth factor (PDGF) and transforming growth factor (TGF)-beta, have been implicated in the pathogenesis of asbestos-induced disease. PDGF and TGF-beta levels were determined by enzyme linked immunosorbent assays in the banked serum samples of a cohort of workers with asbestosis, and the relationships of the growth factor levels to the subsequent development of cancer and to the radiographic severity and progression of asbestosis in the cohort were examined. Serum levels of PDGF and TGF-beta were found to be unrelated to the development of cancer, and serum levels of PDGF were found to be unrelated to the severity and progression of asbestosis. However, serum levels of TGF-beta were found to be statistically significantly related to disease severity (p = 0.01), increasing approximately 2.4-fold from ILO radiographic category 0 to category 3, and they were marginally related to disease progression (p = 0.07), in multivariate analysis controlling for other contributory factors including cumulative asbestos exposure. This suggests that serum TGF-beta may be a useful biomarker for asbestos-induced fibrotic disease. PMID- 19283527 TI - Curcumin sensitizes lung cancer cells to cisplatin-induced apoptosis through superoxide anion-mediated Bcl-2 degradation. AB - The purpose of this study was to investigate the sensitizing effect of curcumin on cisplatin-induced apoptosis in non-small cell lung cancer (NSCLC) H460 cells. Curcumin was shown to induce superoxide anion generation, down-regulate anti apoptotic Bcl-2 protein, and subsequently sensitize cells to cisplatin-induced apoptosis. Co-treatment of the cells with curcumin and cisplatin resulted in increased apoptosis and reversal of Bcl-2-mediated cisplatin resistance. The mechanism by which curcumin down-regulates Bcl-2 and sensitizes cells to cisplatin-induced apoptosis involves proteasomal degradation of Bcl-2. These findings indicate a novel pathway for curcumin regulation of Bcl-2, which could benefit the development of a cisplatin sensitizing agent. PMID- 19283528 TI - A personal account of the rehabilitation system in Japan. PMID- 19283529 TI - Field trial of ICF version for children and youth (ICF-CY) in Sweden: logical coherence, developmental issues and clinical use. AB - INTRODUCTION: For ICF-CY to be used in clinical practice several issues have to be resolved concerning the logical coherence in the model, developmental and functional issues as well as clinical feasibility. PURPOSE: The aim is to investigate feasibility of ICF-CY as expressed by ICF-CY questionnaires in assessment prior to intervention. METHODS: One hundred and thirty-nine professionals working with intervention for children with disabilities used ICF CY questionnaires in assessment. The professionals rated and commented on the feasibility of ICF-CY and ICF-CY questionnaire. Logical coherence, developmental issues and clinical use were investigated. RESULTS: The logical coherence on component level was good (KMO = 0.126, p < 0.005). To investigate developmental and functional issues in the model children, four age groups were compared with the help of one-way ANOVA. Significant differences were found between children younger than 3 and children 3-6 and 7-12 in four chapters in the component Activities/Participation. Professionals consider that ICF-CY is feasible in intervention but experience difficulties when using the components and qualifiers in the ICF-CY and suggest changes. DISCUSSION: ICF-CY as expressed in ICF-CY questionnaires is a feasible tool in assessment and intervention. However, several issues can be developed and discussed to facilitate implementation of ICF CY in clinical practice. PMID- 19283530 TI - Predicting social and functional outcomes for individuals sustaining paediatric traumatic brain injury. AB - OBJECTIVE: To determine the efficacy of models employing the classification of severity of injury, based on either GCS scores or clinical findings using standardized test scores for Occupational Therapy, Physiotherapy and Psychology in predicting outcomes associated with paediatric traumatic brain injury (TBI). METHOD: Medical records were reviewed to obtain GCS scores and standardized tests used in the clinically based classification of severity of injury for 30 individuals who sustained a paediatric TBI and were patients of a brain injury treatment programme. Interviews were conducted with parents to obtain current data on social participation, cognitive functioning and environmental factors. RESULTS: Three variables emerged as significant predictors of outcome: age at injury, clinical ratings of injury severity and environmental factors. CONCLUSION: Findings offer preliminary support for the idea that a combination of factors, including age at injury, clinical expertise and the environment, provide the best estimate of long-term outcome. PMID- 19283531 TI - Technology-based intervention options for post-coma persons with minimally conscious state and pervasive motor disabilities. AB - BACKGROUND: Intervention strategies, based on learning principles and assistive technology, were assessed with four post-coma persons with minimally conscious state and pervasive motor disabilities. METHOD: The first study taught a man to access environmental stimulation through a response-microswitch combination and another man to access environmental stimulation and request social contact through responses combined with a microswitch or a Voice Output Communication Aid (VOCA). The second study taught a man to access two forms of environmental stimulation via two response-microswitch combinations and another man to request two forms of contact via two response-VOCA combinations. RESULTS: Data showed that all participants had significant increases in response levels (independent of whether the responses were combined with microswitch or VOCA devices) during the intervention phases of the studies. CONCLUSION: Intervention strategies based on learning principles and technology may be largely helpful for persons with minimally conscious state and pervasive motor disabilities. PMID- 19283532 TI - Virtual enriched environments in paediatric neuropsychological rehabilitation following traumatic brain injury: Feasibility, benefits and challenges. AB - A frequent consequence of traumatic brain injury (TBI) is a significant reduction in patients' cerebral activation/arousal, which clinicians agree is not conducive to optimal rehabilitation outcomes. In the context of paediatric rehabilitation, sustained periods of inactivity are particularly undesirable, as contemporary research has increasingly called into question the Kennard principle that youth inherently promotes greater neural plasticity and functional recovery following TBI. Therefore, the onus to create rehabilitation conditions most conducive to harnessing plasticity falls squarely on the shoulders of clinicians. Having noted the efficacy of environmental enrichment in promoting neural plasticity and positive functional outcomes in the animal literature, some researchers have suggested that the emerging technology of Virtual Reality (VR) could provide the means to increase patients' cerebral activation levels via the use of enriched Virtual Environments (VEs). However, 10 years on, this intuitively appealing concept has received almost no attention from researchers and clinicians alike. This paper overviews recent research on the benefits of enriched environments in the injured brain and identifies the potential and challenges associated with implementing VR-based enrichment in paediatric neuropsychological rehabilitation. PMID- 19283533 TI - Virtual reality-based paediatric interactive therapy system (PITS) for improvement of arm and hand function in children with motor impairment--a pilot study. AB - OBJECTIVE: Rehabilitation of upper-limb sensorimotor function in children with motor dysfunctions is primarily based on movement training. This study developed a virtual-reality based, paediatric interactive therapy system (PITS) that allows children to practice specific movements of the upper limbs with immediate feedback about their motor performance. METHODS: The system was tested on five children with motor dysfunctions over 3 weeks of training. Pre- and post assessment was conducted before and after the training period. RESULTS: Results of the pilot study show improvements of hand function in the test scores (except one patient). Patient motivation was high and maintained over the course of the therapy sessions. CONCLUSION: PITS is an applicable VR-system which can be feasibly applied during the rehabilitation of children with upper limb motor dysfunctions. Further investigation is necessary to determine if the system provides significantly improved results compared to conventional therapies, both in terms of motor function outcomes and patient motivation. PMID- 19283535 TI - Health competent societies: our challenge and future. PMID- 19283536 TI - Usability testing by older adults of a computer-mediated health communication program. AB - Failure to adhere to an antihypertensive regimen and interactions between antihypertensives and other medicines represent serious health threats to older adults. This study tested the usability of a touch-screen-enabled personal education program (PEP). Findings showed that older adults rated the PEP system usability, system usefulness, and system-use satisfaction at a moderately high level for prototype-1 and at an exceptionally high level for prototype-2. A 201.91% reduction in interface errors and a 31.08% decrease in interface time also were found between the two trials. This participatory usability design was highly successful in tailoring its program interface design to accommodate older users to enhance their health communication and technology use efficacy. PMID- 19283537 TI - Practicing what they preach: health behaviors of those who provide health advice to extensive social networks. AB - As a way of identifying a conduit to disseminate health information, this study aims to explore health behaviors and attitudes of a unique group of extensively socially-networked individuals who regularly are asked for their health advice. Respondents from a population-based consumer opinion panel (n = 2,639) were categorized as "extensively socially-networked" (75+ friends and acquaintances, and almost daily giving friends advice on general issues) vs. "non-networked." The networked respondents were further divided into "health-networked" (regularly asked for health advice) versus "only-socially-networked" groups (asked for general advice, not health). Chi-square analyses, ANOVA tests, and multivariate regressions controlling for sociodemographic variables compared health behaviors and attitudes between groups. Results indicated that health-networked individuals reported more positive health behaviors (e.g., fruit and vegetable consumption) and attitudes than only-socially-networked and non-networked individuals. Future research is warranted to elucidate how providing health advice to a large network contributes to the positive health of health-networked individuals. Exploratory analyses revealed that doctors and health/fitness magazines were main sources of health and nutrition information for health-networked respondents. Through their advice and word-of-mouth, health-networked individuals have the potential to influence the health information of large groups of people and, therefore, may serve as valuable change agents to disseminate health and nutrition information. PMID- 19283538 TI - Developing effective campaign messages to prevent neural tube defects: a qualitative assessment of women's reactions to advertising concepts. AB - The incidence of neural tube defects (NTDs), serious birth defects of the brain and spine that affect approximately 3,000 pregnancies in the United States each year, can be reduced by 50-70% with daily periconceptional consumption of the B vitamin folic acid. Two studies were designed to assess college women's reactions to and perceptions of potential campaign advertising concepts derived from preproduction formative research to increase folic acid consumption through the use of a daily multivitamin. Study one assessed draft advertising concepts in eight focus groups (N = 71) composed of college-enrolled women in four cities geographically dispersed across the United States. Based on study one results, the concepts were revised and reassessed in study two with a different sample (eight focus groups; N = 73) of college women in the same four cities. Results indicated that participants generally responded favorably to concepts in each of the two studies, and provided insight into individual concepts to increase their overall appeal and effectiveness. The specific findings and implications of these results are discussed. PMID- 19283539 TI - Exploring antecedents of consumer satisfaction and repeated search behavior on e health information. AB - E-health information has become an important resource for people seeking health information. Even though many studies have been conducted to examine the quality of e-health information, only a few studies have explored the effects of the information seekers' motivations on the perceived quality of e-health information. There is even less information about repeated searches for e-health information after the users' initial experience of e-health information use. Using an online survey of information seekers, 252 e-health information users' responses were collected. The research examines the relationship among motivation, perceived quality, satisfaction, and intention to repeat-search e health information. The results identify motivations to search e-health information and confirm the relationship among motivation, perceived quality dimensions, and satisfaction and intention to repeat searches for e-health information. PMID- 19283540 TI - Break it to me harshly: the effects of intersecting news frames in lung cancer and obesity coverage. AB - By examining the publicily identified top two health problems in the United States, this research, using an experimental design, investigates whether different news frame combinations intensify or diminish framing effects. In this study, the cognitive dimension and affective dimension of framing defined as thematic/episodic and gain/loss, respectively, are manipulated to determine if changing the way newspaper stories report obesity and lung cancer will alter the readers' attribution of societal and individual responsibility. This study revealed a significant interaction between thematic framing and loss framing on societal attribution of responsibility for the health issues-lung cancer and obesity. PMID- 19283541 TI - Black youth's personal involvement in the HIV/AIDS issue: does the public service announcement still work? AB - Recent public service announcements (PSAs) directed toward Black youth utilize various formats and appeals to stimulate a motivated cognitive process that engenders personal involvement in the HIV/AIDS issue. The Elaboration Likelihood Model (ELM) by Petty and Cacioppo argues that engagement with messages that consist of substantive content causes the audience member to critically analyze the message, which can produce awareness and attitude change. An efficient way to add emphasis to the message and seize the attention of the target audience is to insert the message into an entertainment context. Our study attempted to analyze the impact of the peripheral cue, character appeal, on audience members' attitude change in response to analyzing high- and low-involvement message content. A2 x 4 factorial design was used, with message involvement (high/low) and character appeal (White/Black and celebrity/noncelebrity) as independent variables. The findings showed that celebrity status is the salient factor, with source perception inducing attitude change as a main effect or in an interaction effect with high- and low message content. PMID- 19283544 TI - The choices, choosing model of quality of life: clinical evaluation and intervention. AB - Two previous papers presented a conjectured model of quality of life featuring the personal management of accessing choices and choosing among them (the c-c process). Those papers made the case that this model unifies the pathways leading to impaired quality of life, especially with regard to age associated multiple co morbidities, changes in perception and functioning, and the effects of long term care environments; it also introduces a science base for understanding and guiding interventions that can assist people to achieve their quality-of-life goals. Our aim in this paper is to outline interprofessional strategies that could relieve restrictions or distortions of the c-c process imposed by aging, ill-health, or a restricting environment. We do so by outlining potential deficits in the c-c process and matching these with restorative person-centered interprofessional interventions including interprofessional teamwork. Findings suggest that interprofessional assessment and team work is well suited to assisting the c-c process. We conclude that the groundwork has been prepared for developing training programs and clinical trials for interprofessional interventions targeting the c-c process. PMID- 19283545 TI - Innovation in collaborative health research training: the role of active learning. AB - This paper describes and discusses the essential pedagogical elements of the Partnering in Community Health Research (PCHR) program, which was designed to address the training needs of researchers who participate in collaborative, interdisciplinary health research. These elements were intended to foster specific skills that helped learners develop research partnerships featuring knowledge, capabilities, values and attitudes needed for successful research projects. By establishing research teams called "clusters", PCHR provided research training and experience for graduate students and post-doctoral fellows, as well as for community health workers and professionals. Pedagogical elements relied on active learning approaches such as inquiry-based and experience-based learning. Links between these elements and learning approaches are explained. Through their work in cluster-based applied research projects, the development of learning plans, and cross-cluster learning events, trainees acquired collaborative research competencies that were valuable, relevant and theoretically informed. PMID- 19283546 TI - Enhancing research interest and collaboration in the interdisciplinary context of emergency care. AB - The interdisciplinary context of the emergency department encompasses diverse clinical presentations requiring teamwork by doctors, nurses and allied health workers to achieve optimal patient care. This interdisciplinary focus is extended by adding a research perspective. This project sought to systematically examine the current research capacity of emergency department staff at a major Australian tertiary urban hospital and to derive information about further research-related needs with a view to enhancing research capacity. The mixed method project utilized a department-wide staff survey followed by focus groups and individual interviews. Adequate response rates to the two phases were achieved (n = 67, n = 17 respectively). Not surprisingly, 89% of participants reported that they needed help with developing their research skills. Clinicians reported little or no experience with (i) finding literature (35%) and critical review (50%), (ii) research skills and techniques, both qualitative (72%) and quantitative (63%), and (iii) research output: publishing (68%), writing & presenting (34%). Data from focus groups and individual interviews yielded themes around developing research skills, communication, meaningfulness, team work and interdisciplinary strategies, forming part of the Dimensional Enhancing Research Capacity (DERC) model. This project highlighted not only interdisciplinary needs for research but also the way that research may additionally assist with building interprofessional linkage. PMID- 19283550 TI - Relationships between self-evaluations of voice and working conditions, background factors, and phoniatric findings in female teachers. AB - Self-evaluations of voice, working conditions, and results of phoniatric examination were studied in 78 female primary school teachers. Most teachers evaluated their vocal capabilities as 'normal'; nevertheless 1/3 reported two or more weekly symptoms. Unsatisfactory air quality, background noise, and stressful working conditions were seen as the most harmful environmental factors. A total of 14% of the subjects had organic laryngeal changes, 37% had mild changes, and 49% were found to be healthy. These findings did not correlate with self evaluation of voice, vocal symptoms, or voice-related quality of life. Teaching experience or teaching hours per week did not correlate with laryngeal findings or symptoms. The results illustrate the complex nature of voice assessment and the need of assessment tools specially suited for functionally healthy voice professionals. PMID- 19283551 TI - Perception and suppression of thermally induced pain: a fMRI study. AB - Two neuroimaging studies using functional magnetic resonance imaging (fMRI) and thermally induced pain are presented. Fifteen healthy right-handed subjects were imaged while they had to discern different levels of thermal stimuli in the first study and while they disengaged from the feeling of pain during constant stimulation in the second study. In the first experiment, during painful phasic stimuli, right-sided anterior insular activation as well as bilateral posterior insular activation could be shown regardless of stimulation side, as well as right-sided activation of sensory association areas in the superior parietal lobule. Also, activation of the ipsilateral sensorimotor cortex could be shown. In the second experiment, all subjects succeeded in suppressing the feeling of pain during previously painful levels of stimulation. During the early part of the tonic painful stimulation, bilateral activation of caudate head and dorsolateral prefrontal cortex (DLPFC) as well as insular cortex and dorsal anterior cingulated cortex (dACC) was observed. During the late part of the tonic painful stimulation, anterior insular activation as well as dACC and bilateral prefrontal cortical activation could be shown. Taken together, the activation of PFC and caudate nucleus hints at an important role in the initiation (caudate) and maintenance (PFC) of suppression of the feeling of pain. No ipsilateral sensorimotor activation could be shown in the second experiment. The possible import of unwanted sensorimotor activation due to the simultaneous rating process in the first experiment is discussed. PMID- 19283552 TI - Independent psychophysical measurement of experimental modulations in the somatotopy of cutaneous heat-pain stimuli. AB - Distortions of the body image have been repeatedly reported for various clinical conditions, but direct experimental analyses of the perceptual changes involved are still scarce. In addition, most experimental studies rely on cerebral activation patterns to assess neuroplastic changes in central representation, although the relationship between cerebral topography and the topology of the perceptual space is not clear. This study examines whether the direct psychophysical mapping approach we introduced recently (Trojan et al., Brain Res 2006;1120:106-113) is capable of tracking perceptual distortions in the somatotopic representation of heat-pain stimuli. Eleven healthy participants indicated the perceived positions of CO(2) laser stimuli, repetitively presented to the dorsal forearm, with a 3D tracking system in two consecutive sessions, separated by the topical application of capsaicin cream. In line with earlier reports, we expected that the resulting individual perceptual maps (i.e., one dimensional projections of the perceived positions onto the forearm surface) would be subject to modulation through the altered sensory input, to be measured in terms of altered topological parameters. We found that the topology and metrics of the somatotopic representation were well preserved in the second session, but that the perceptual map was compressed to a smaller range in 9 out of 11 participants. By providing dimensional measures of perceptual representations, perceptual maps constitute an independent, genuinely psychological complement to the topography of cortical activations measured with neuroimaging methods. In addition, we expect them to be useful in diagnosing pathological changes in body perception accompanying chronic pain and other disorders. PMID- 19283553 TI - Wally Welker 1926-2007. PMID- 19283554 TI - Barrels come of age: Barrels XXI meeting report. AB - The twenty-first annual Barrels meeting, sponsored by NINDS, was held on 12-14 November 2008 on the campus of Johns Hopkins University, near the site of the original discovery of barrels almost 40 years ago. The longest running satellite meeting to the Society for Neuroscience Annual Meeting focuses on the development, physiology, and behavior of the rodent whisker-to-barrel sensorimotor system. This year's event focused on what aspects of the sensory world are encoded by neurons within the system and how specifically the posterior medial nucleus can play a role in information processing. Other highlighted topics included the possible role(s) the cerebellum may have and the cues governing the patterning and development of thalamocortical inputs into the barrel cortex. PMID- 19283555 TI - Implicit motor sequence learning is not represented purely in response locations. AB - This study employed a novel variant of the serial reaction time task, focused on sequencing one element of movement-direction. During the task a repeated pattern of alternating directions (right-left-right, etc.) was embedded in the stimuli, and there was no series of response locations. Responses were made via two effector systems: single-finger responding (necessitates lateral arm movements between response keys), and four-fingered responding (4 individual fingers on 4 individual keys; requires no lateral arm movement). The sequence of directions was only learned by participants who performed lateral movements during training, indicating that learning was contingent on the particular motor effector used. Participants with low levels of sequence awareness displayed the same pattern of results. PMID- 19283556 TI - Phonological neighbourhood spread facilitates lexical decisions. AB - In visual word recognition, words with many phonological neighbours are processed more rapidly than are those with few neighbours. The research reported here tested whether the distribution of phonological neighbours across phoneme positions influences lexical decisions. The results indicate that participants responded more rapidly to words where all phoneme positions can be changed to form a neighbour than they did to those where only a limited number of phoneme positions can be changed to form a neighbour. It is argued that this distribution effect arises because of differences between the two groups of words in how they overlap with their neighbours. PMID- 19283557 TI - Verb aspect and perceptual simulations. AB - Two experiments investigate the influence of verb aspect on situation representations. The results demonstrate that comprehenders use verb aspect as a cue to regulate the activation of ongoing simulations of situations over time. Experiment 1 measured word-by-word reading as well as sensibility judgements on sentences in which a target object word had been replaced by a picture. For the past imperfective sentences, participants were faster to process the picture, the two words following the picture, and the sensibility judgements when objects were pictured in use rather than not in use. However, this in-use facilitation was limited to processing of the picture for the past perfect sentences. Experiment 2 served as a control to ensure that the use effect and its interaction with verb aspect were a result of contextual manipulations rather than surface features of the pictures themselves. The results are interpreted within the framework of perceptual simulations during language comprehension. PMID- 19283558 TI - Adult age differences in memory for distinctive information: evidence from the bizarreness effect. AB - Two experiments examined age-related differences in memory for bizarre and common pictures. In Experiment 1, a facilitative effect of bizarreness was obtained for young adults and one of the older groups, but not for the oldest group (over age 70). However, the bizarreness effect was found for even the oldest group when predominantly common lists were used in Experiment 2. It is concluded that older adults suffer from deficits in distinctive processing, but those deficits can be reduced by providing a more uniformly common context in which differences can be processed. PMID- 19283559 TI - Early detection and avoidance of threatening faces during passive viewing. AB - To evaluate whether there is an early attentional bias towards negative stimuli, we tracked participants' eyes while they passively viewed displays composed of four Ekman faces. In Experiment 1 each display consisted of three neutral faces and one face depicting fear or happiness. In half of the trials, all faces were inverted. Although the passive viewing task should have been very sensitive to attentional biases, we found no evidence that overt attention was biased towards fearful faces. Instead, people tended to actively avoid looking at the fearful face. This avoidance was evident very early in scene viewing, suggesting that the threat associated with the faces was evaluated rapidly. Experiment 2 replicated this effect and extended it to angry faces. In sum, our data suggest that negative facial expressions are rapidly analysed and influence visual scanning, but, rather than attract attention, such faces are actively avoided. PMID- 19283560 TI - Tobacco use by physicians in a physician health program, implications for treatment and monitoring. AB - The use of tobacco by physicians with substance abuse histories is drastically understudied. A chart review of 1319 physicians enrolled in a physician health program found tobacco use highest for those referred for substance abuse problems (58.1%). Among a subset of currently monitored substance abusers, all those who relapsed during monitoring were using tobacco and had more difficulty maintaining sobriety following initial treatment (p = 0.0137) than non tobacco users. Because tobacco was a risk factor for relapse, reasons why physician health programs should address its use and treatment facilities should establish tobacco-free environments to provide optimum learning and recovery are explored. PMID- 19283561 TI - Early outcomes following low dose naltrexone enhancement of opioid detoxification. AB - Although withdrawal severity and treatment completion are the initial focus of opioid detoxification, post-detoxification outcome better defines effective interventions. Very low dose naltrexone (VLNTX) in addition to methadone taper was recently associated with attenuated withdrawal intensity during detoxification. We describe the results of a seven-day follow-up evaluation of 96 subjects who completed inpatient detoxification consisting of the addition of VLNTX (0.125 or 0.250 mg per day) or placebo to methadone taper in a double blind, randomized investigation. Individuals receiving VLNTX during detoxification reported reduced withdrawal and drug use during the first 24 hours after discharge. VLNTX addition was also associated with higher rates of negative drug tests for opioids and cannabis and increased engagement in outpatient treatment after one week. Further studies are needed to test the utility of this approach in easing the transition from detoxification to various follow-up treatment modalities designed to address opioid dependence. PMID- 19283562 TI - Pain and substance-related pain-reduction behaviors among opioid dependent individuals seeking methadone maintenance treatment. AB - We surveyed 293 opioid dependent individuals seeking methadone maintenance treatment about their pain experiences and their substance-related pain reduction behaviors. Among the 213 respondents reporting recent pain of at least moderate typical pain intensity, two-thirds had a lifetime history of chronic pain. In comparison to those without a lifetime history of chronic pain, those with a lifetime history were older, reported higher pain frequency, were more likely to endorse accident or surgery and less likely to endorse "don't know" as the genesis of their recent pain, and endorsed comparable levels of substance-related pain reduction behaviors. These findings may have implications for resource and program planning in MMT programs. PMID- 19283563 TI - Using vouchers to increase continuing care participation in veterans: does magnitude matter? AB - The present study examined the comparative effects of adding contingency management (CM) schedules to an existing substance abuse continuing care program, with the goal of increasing attendance. We retrospectively examined the attendance of 135 veterans enrolled in one of three CM programs and a quasi control condition of 55 veterans. Results indicated that participants enrolled in the two higher magnitude CM voucher programs increased both continuing care attendance and length of participation. Findings support the use of CM to increase continuing care attendance among veterans with substance use disorders, and suggest that voucher magnitude and bonuses both had a positive impact. PMID- 19283564 TI - Evening experiences versus drinking indicators as predictors of hangover on a summer holiday. AB - Little is known about predictors of hangover symptoms. The subjective experience of the "night out" may influence the experience of hangover symptoms. We conducted a survey of 325 young men and women at a vacation resort during the daytime, administering subjects the Acute Hangover Scale (AHS) and asking questions about drinking and behaviors the night before. Half of the subjects reported feeling hangover symptoms on a single item relating to hangovers in general, but 73% scored above 1 on the full AHS. Several variables correlated with AHS, but in multivariate analyses, only variables associated with drunkenness the night before (having been more drunk than intended and still feeling intoxicated) and number of days spent in the resort were associated with AHS. Future studies should work on developing standardized definitions of hangover that can be used to study the phenomenon further. PMID- 19283565 TI - Characteristics and outcomes of patients discharged from the Opioid Renewal Clinic at the Philadelphia VA Medical Center. AB - The Opioid Renewal Clinic (ORC) is a pharmacist-run program that assists primary care providers in the management of chronic pain patients with aberrant behavior or high risk of prescription opioid abuse. In this study, charts of all discharged patients over a 22-month span (n = 86) were reviewed for outcomes for two years after discharge. The most frequent reason for discharge from the program was recurrent positive urine drug screens for illicit substances (n = 40; 47%). Only 15 (17%) were seen for addiction treatment in the two-year follow-up period. Forty-one percent of patients (n = 35) were prescribed opioids within two years of discharge. There was a positive correlation between length of time in the ORC and receipt of opioids post discharge. These outcomes reveal areas of need in the management of this complex population. PMID- 19283566 TI - Gender differences in associations between lifetime alcohol, depression, panic disorder, and posttraumatic stress disorder and tobacco withdrawal. AB - This study examined the interaction of gender and lifetime psychiatric status on the experience of nicotine withdrawal using retrospective data from the National Comorbidity Survey (NCS; N = 816). Multiple regression analyses were performed to examine the main and interactive effects of gender and major depression, alcohol abuse/dependence, panic disorder, and PTSD on indices of withdrawal. Major depression and alcohol abuse/dependence were associated with longer duration of withdrawal symptoms in women. Women also showed stronger associations between major depression and recurrent withdrawal symptoms and PTSD and smoking relapse to alleviate withdrawal. Men showed a stronger association between alcohol abuse/dependence and smoking relapse to alleviate withdrawal. When developing and providing smoking cessation interventions, it is important to consider the gender specific effects of lifetime psychiatric status on withdrawal. PMID- 19283568 TI - Empathy ability is impaired in alcohol-dependent patients. AB - Empathy is a complex form of psychological inference in which observation, memory, knowledge and reasoning are combined to yield insights into the thoughts and feelings of others. The aim of this study was to evaluate the level of empathy in a sample of alcohol-dependent patients in comparison to a control sample. One hundred and fifty alcohol-dependent subjects were consecutively recruited. All of the subjects successfully detoxified have been evaluated with the Empathy Quotient (EQ) and then compared with 107 control subjects. The level of empathy was significantly lower in the group of alcohol-dependent subjects than in the control sample (p <.001). Differences with respect to gender and psychiatric comorbidity have also been observed. A low level of empathy could be a psychological trait typically observed in pre-morbid alcoholic personalities. Further, the lack of empathy could lead latent abusers to find in the alcohol misuse something enabling them to compensate for their intrinsic weakness. PMID- 19283567 TI - Psychopharmacologic management of opioid-dependent women during pregnancy. AB - Illicit drug use during pregnancy presents complex clinical challenges, including reducing drug use and treating psychiatric disorders. Pharmacologic treatment of psychiatric disorders in a pregnant woman requires an evaluation of the balance between potential clinical benefit and the risk of potential neonatal consequences. This study describes psychiatric symptoms in 111 opioid-dependent pregnant women and their prescribed psychotropic medications. Hypomania, generalized anxiety disorder and depression were the most common disorders for which psychiatric symptoms were endorsed. Over half of women studied were prescribed some form of psychoactive medication during pregnancy. Pharmacologic vs. non-pharmacologic treatment approaches in this patient population are discussed. PMID- 19283569 TI - Misuse of alcohol among methadone patients. AB - Between 5 and 49% of patients undergoing methadone maintenance treatment (MMT) consume alcohol. They show less willingness to comply with treatment and are less committed, their treatment times are longer, and their observance is weaker. Alcohol misuse is the main cause of their increased mortality and morbidity rates. We conducted a multivariate, prospective, open-label study in two groups of methadone patients: the first suffered from alcohol use disorder according to the criteria of the alcohol use disorder test (AUDIT), while the second was alcohol-abstinent. In the process, we completed evaluations on the Toronto alexithymia scale, together with a self-esteem inventory and a temperament and character inventory (TCI-R). Our study included 152 patients, 27.6% of whom were high-risk consumers. Comparison of the alcohol high-risk consumers group and the low-risk consumers showed that the former had higher rates of alexithymia, weaker self-esteem, and lower scores of self-directedness. PMID- 19283570 TI - The influence of intake urinalysis, psychopathology measures, and menstrual cycle phase on treatment compliance. AB - Drug abuse among women is a societal health problem which has received greater attention in recent years. Studies examining sex differences in drug abuse have shown consistent differences between males and females. In the current study, we retrospectively examined 75 treatment-seeking females to determine the relationships between intake measurements of psychopathology, intake urinalysis, menstrual cycle phase at admission, and treatment compliance. Our results support a relationship among these variables and a potential relationship between menstrual cycle phase and anxiety level is discussed. These results support the need for the development of individually tailored treatment programs. PMID- 19283571 TI - The Violence Proneness Scale of the DUSI-R predicts adverse outcomes associated with substance abuse. AB - Accuracy of the Violence Proneness Scale (VPS) of the Drug Use Screening Inventory (DUSI-R) was evaluated in 328 boys for predicting use of illegal drugs, DUI, selling drugs, sexually transmitted disease, car accident while under acute effects of drugs/alcohol, trading drugs for sex, injuries from a fight, and traumatic head injury. Boys were prospectively tracked from age 16 to 19 at which time these outcomes were documented in the interim period. The results demonstrated that the VPS score is a significant predictor of all outcomes. Prediction accuracy ranged between 62%-83%. These findings suggest that the VPS may be useful for identifying youths who are at high risk for using illicit drugs and commonly associated adverse outcomes. PMID- 19283572 TI - Challenges in quantifying marijuana use. PMID- 19283573 TI - Is there a better way to match patients to opioid maintenance treatment: a case report. PMID- 19283574 TI - Acceptability of technology-based methods substance abuse counseling in office based buprenorphine maintenance for opioid dependence. PMID- 19283575 TI - Auditory processing in speakers with acquired or developmental language disorders. Preface. PMID- 19283576 TI - Understanding the role of the prefrontal cortex in phonological processing. AB - Lesion studies have demonstrated impairments of specific types of phonological processes. However, results from neuropsychological studies of speech sound processing have been inconclusive as to the role of specific brain regions because of a lack of a one-to-one correspondence between behavioural patterns and lesion location. Functional neuroimaging studies have contributed more detailed information about the involvement of specific brain regions in a wide range of phonological tasks. A framework developed by Hickok and Poeppel to account for these neuropsychological and neuroimaging results is evaluated in light of a series of phonological studies in which cognitive load is manipulated by changing the acoustic properties and lexical status of stimuli, as well as the type of phonological judgement. Overall, the findings for speech stimuli are consistent with the view that tasks that require increased articulatory recoding result in increased activation of the posterior aspect of the inferior frontal gyrus (BA 44). However, similar activation patterns for tone sequences as compared to speech may challenge whether the recoding is speech-specific. Implications of these investigations for future neuroimaging studies of individuals with aphasia are discussed. PMID- 19283577 TI - Neighbourhood density effects in auditory non-word processing in aphasic listeners. AB - This study investigates neighbourhood density effects on lexical decision performance (both accuracy and response times) of aphasic patients. Given earlier results on lexical activation and deactivation in Broca's and Wernicke's aphasia, the prediction was that smaller neighbourhood density effects would be found for Broca's aphasic patients, compared to age-matched non-brain-damaged control participants, whereas enlarged density effects were expected for Wernicke's aphasic patients. The results showed density effects for all three groups of listeners, and overall differences in performance between groups, but no significant interaction between neighbourhood density and listener group. Several factors are discussed to account for the present results. PMID- 19283578 TI - The influence of visual and auditory information on the perception of speech and non-speech oral movements in patients with left hemisphere lesions. AB - Patients with lesions of the left hemisphere often suffer from oral-facial apraxia, apraxia of speech, and aphasia. In these patients, visual features often play a critical role in speech and language therapy, when pictured lip shapes or the therapist's visible mouth movements are used to facilitate speech production and articulation. This demands audiovisual processing both in speech and language treatment and in the diagnosis of oral-facial apraxia. The purpose of this study was to investigate differences in audiovisual perception of speech as compared to non-speech oral gestures. Bimodal and unimodal speech and non-speech items were used and additionally discordant stimuli constructed, which were presented for imitation. This study examined a group of healthy volunteers and a group of patients with lesions of the left hemisphere. Patients made substantially more errors than controls, but the factors influencing imitation accuracy were more or less the same in both groups. Error analyses in both groups suggested different types of representations for speech as compared to the non-speech domain, with speech having a stronger weight on the auditory modality and non-speech processing on the visual modality. Additionally, this study was able to show that the McGurk effect is not limited to speech. PMID- 19283579 TI - Speech perception in children with speech output disorders. AB - Research in the field of speech production pathology is dominated by describing deficits in output. However, perceptual problems might underlie, precede, or interact with production disorders. The present study hypothesizes that the level of the production disorders is linked to level of perception disorders, thus lower-order production problems (such as childhood apraxia of speech; CAS) are linked to lower-order perception problems and higher-order production problems (phonological disorder; PD) are linked to higher-order perception problems. For this, various perception tasks were administered (non-word auditory discrimination task, word rhyming task, categorical classification, and discrimination task) in children with CAS, PD, or a mixture. The results show that children with PD only show higher-order perception problems, whereas children with CAS have difficulties on both lower- and higher-order perception tasks. In children with CAS, difficulties at lower-order processes might affect the higher-order processes in development. Furthermore, significant correlations were found between production and perception scores at different levels. Thus, a link between perception and production seems to be evident. PMID- 19283580 TI - IJA's impact on world-wide audiology. PMID- 19283581 TI - Attitude and knowledge of hearing loss among medical doctors selected to initiate a residency in Mexico. AB - Early diagnosis and intervention of hearing loss are directly influenced by the knowledge and attitude towards this condition among medical personnel, particularly in countries where screening is not performed routinely. The objective of this paper was to evaluate the attitude and knowledge of hearing loss in a group of physicians. A questionnaire with five Likert-type items and five multiple choice and fill-in-the-blank items was completed by 2727 physicians selected to start a medical residency. Results suggested that physicians' knowledge level on the matter is deficient and their attitude is far from the ideal; however, physicians selected for a residency in audiology showed slightly better results. PMID- 19283582 TI - An implementation study of neonatal hearing screening in the Netherlands. AB - The objective of this study was to investigate whether universal neonatal hearing screening could be integrated in the youth health care program. The screening was performed by nurses of the well baby clinics. A three stage transient evoked otoacoustic emission screening was performed in three different screening settings in order to study the most effective set up regarding participation, refer rates, and costs. In one setting parents visited the well baby clinic, and in two settings babies were screened at home (either in combination with the screening for metabolic diseases or during an intake visit). Screening was performed on 3114 healthy newborns. The setting where universal neonatal hearing screening is integrated with the screening for metabolic diseases, proved to be most efficient and effective. The participation rate of 88.9% was highest in this setting and the overall refer rate (1.4%) was the lowest. The implementation of universal neonatal hearing screening by the well baby clinic nurses was judged to be possible. The results of this study formed the basis for nationwide implementation. PMID- 19283583 TI - Hearing loss and paid employment: Australian population survey findings. AB - This paper provides an analysis of participation in paid employment for people with a hearing loss over the full span of adult ages. The paper is based on original analysis of the 2003 Australian survey of disability, aging and carers (SDAC). This analysis shows that hearing loss was associated with an increased rate of non-participation in employment of between 11.3% and 16.6%. Advancing age and the existence of co-morbidities contribute significantly to reduced participation in employment. A disproportionate impact is evident for women and for those having low education and communication difficulties. Controlling for co morbidities, hearing loss was associated with a 2.1% increase of non participation in employment, a proportional difference of 1.4 times the population. People with hearing loss were less likely to be found in highly skilled jobs and were over-represented among low income earners. The SDAC data set provides self-report findings on the experience of disability rather than hearing impairment. As such, these findings serve as a conservative estimate of the impact of hearing loss on accessing well-paid employment. PMID- 19283584 TI - The extent of hearing impairment amongst Australian Indigenous prisoners in Victoria, and implications for the correctional system. AB - The hearing status of 109 Indigenous prisoners was investigated at five prison locations in Victoria, using audiological methods and face-to-face interview. The study found predominantly mild, sensorineural hearing loss. The rate of conductive hearing impairment was consistent with an age-matched general adult population (UK). All eardrums were intact, and 89% of middle-ears were normally air filled. Results showed 12% of prisoners had a hearing loss (average. 0.5, 1, 2, & 4 kHz >or=25 dB) in at least one ear, compared with 5% in an age-matched Australian adult population. More than a third (36%) had high-frequency, sensorineural hearing impairment (4 or 6 kHz >or=25 dB), in one or both ears. Over half of the inmates (58%) reported hearing problems sometimes, and 4% reported a lot of hearing trouble. The majority of prisoners (92%) reported exposures to loud noise, and tinnitus was reported by 72% of prisoners. For hearing-impaired individuals within the correctional system, the reduced ability to communicate with ease may impact detrimentally on daily interactions, and may impede progress through rehabilitation programs. PMID- 19283585 TI - Auditory disorders in a South African paediatric TBI population: some preliminary data. AB - South Africa has an exceptionally high incidence of traumatic brain injury in its paediatric population but very little systematic research has been conducted on the outcome of such injury. A number of socio demographic variables influence the mechanism of injury as well as its management. This paper reports on a study documenting the educational and audiological outcomes of a group of 100 subjects who sustained their injuries prior to the age of 12 years. The hearing and communication profile of this group is considered in relation to educational outcome. An illustrative case study will demonstrate the multiple influences on outcome in this setting, as well as the need for integrated team management. In the sample, 31% had a reported hearing loss, confirmed audiologically in 14% of subjects. The majority of the sample returned to a mainstream school. However, analysis indicated that about 75% of the sample was in need of specialized education, while only 22% of that sample had received any additional support. The implications of these findings for audiological policy and practice are highlighted. PMID- 19283586 TI - Cost-effectiveness of selected interventions for hearing impairment in Africa and Asia: a mathematical modelling approach. AB - The purpose of this paper was to present estimates of costs and effects of selected interventions for hearing impairment in Africa and Asia. The method used mathematical simulation models on the basis of WHO burden of disease information, and WHO-CHOICE costing databases. Findings showed that in both regions, screening strategies for hearing impairment and delivery of hearing aids cost between I$1000 and I$1600 per DALY, with passive screening being the most efficient intervention. Active screening at schools and in the community are somewhat less cost-effective. In the treatment of chronic otitis media, aural toilet in combination with topical antibiotics costs is more efficient than aural toilet alone, and costs between I$11 and I$59 in both regions. The treatment of meningitis with ceftriaxone costs between I$55 and I$217 at low coverage levels, in both regions. In more absolute terms, the vast majority of all considered intervention strategies are cost-effective strategies according to international benchmarks, in both regions concerned. In conclusion, various strategies are economically attractive to reduce the disease burden of hearing impairment around the world. PMID- 19283587 TI - Comment on Baltussen and Smith. Cost-effectiveness of selected interventions for hearing impairment in Africa and Asia: a mathematical modelling approach. PMID- 19283588 TI - Peripheral asymmetry of cell-mediated immunity. AB - Recent studies revealed that the peripheral cell-mediated immunity is affected by cerebral motor lateralization in humans. PMID- 19283589 TI - Antiepileptogenic effects of glutathione against increased brain ADA in PTZ induced epilepsy. AB - Adenosine has been shown to play a significant role as a modulator of neuronal activity in convulsive disorders, acting as an endogenous anticonvulsant agent. Any change in adenosine deaminase (ADA) levels will reflect to adenosine levels. In the present study, we have investigated the effect of glutathione on brain tissue ADA levels due to seizures induced by convulsive and subconvulsive dose of pentylenetetrazol (PTZ) in mice. ADA levels due to seizures induced by convulsive and subconvulsive pentylenetetrazol were measured using the Giusti method. ADA levels were higher in the experimental epilepsy groups than in the control and sham groups. ADA levels significantly decreased in the glutathione groups, which may have antiseizure effects. Decreased levels of ADA would be due to increased adenosine levels, protecting against oxidative stress. PMID- 19283590 TI - Moderate pressure massage elicits a parasympathetic nervous system response. AB - Twenty healthy adults were randomly assigned to a moderate pressure or a light pressure massage therapy group, and EKGs were recorded during a 3-min baseline, during the 15-min massage period and during a 3-min postmassage period. EKG data were then used to derive the high frequency (HF), low frequency (LF) components of heart rate variability and the low to high frequency ratio (LF/HF) as noninvasive markers of autonomic nervous system activity. The participants who received the moderate pressure massage exhibited a parasympathetic nervous system response characterized by an increase in HF, suggesting increased vagal efferent activity and a decrease in the LF/HF ratio, suggesting a shift from sympathetic to parasympathetic activity that peaked during the first half of the massage period. On the other hand, those who received the light pressure massage exhibited a sympathetic nervous system response characterized by decreased HF and increased LF/HF. PMID- 19283591 TI - Movement-related cortical evoked potentials using four-limb imagery. AB - We compared the electroencephalographic changes during actual and imaginary movements with four limbs and classified optimally the responses during four-limb imagery. Evoked potentials in imagery exhibited lower and delayed peaks compared to actual-movement responses, but activations in the primary and the supplementary motor area were similar. Source-modeling analysis revealed that the motor and the parietal cortex were activated similarly, but several dipole sources were active in the frontal cortex for imagery. We compared thirteen classification methods and a combination of template matching and time-frequency methods showed the highest average of 70% classification rate for all limbs. PMID- 19283592 TI - Amitriptyline treatment under chronic stress conditions: effect on circulating catecholamines and anxiety in early maternally separated rats. AB - The aim of this work was to determine the effect of amitriptyline (AMI) on peripheral outcomes such as plasma epinephrine (E) and norepinephrine (NE) concentration and anxiety-like behavior displayed in the plus maze test in adult male Wistar rats under variable chronic stress and daily oral administration of AMI (5 mg/kg). Animals were previously isolated from the mother for 4.5 hr every day for the first 3 weeks of life. Administration of the antidepressant AMI reduced anxiety-like behavior in animals submitted only to chronic stress but not in early maternally separated (MS) subjects or in animals subjected to the two types of stresses. PMID- 19283593 TI - The role of mirror neurons in processing vocal emotions: evidence from psychophysiological data. AB - Recent evidence suggests that the mirror neuron system may serve as a common neural substrate for processing motor, linguistic, emotional, and other higher level cognitive information. The current study employed psychophysiological methods to elucidate the role of this system in processing vocal emotions. Skin conductance and heart rate were measured for 25 undergraduate students while they were both listening to emotional vocalizations and also thinking (internal production) about them. The results revealed changes in skin conductance response and heart rate during both "listening" and "thinking" conditions. This suggests an active role of the mirror neuron system in processing vocal emotions. PMID- 19283595 TI - The rabies virus phosphoprotein synthesis and subcellular localization are modified by nerve growth factor. AB - Rabies virus P protein participates as a regulating factor in viral transcription and replication; recent studies found an antitranscriptional and antireplicative effect of nerve growth factor (NGF) and Neurotrophin-3 (NT-3) in infected neuron cultures. We investigated here the specific effect of the neurotrophins on P protein, evaluating its synthesis and subcellular distribution in adult mouse dorsal root ganglia neuron cultures infected and treated with NGF or NT-3. The results showed that NGF, but not NT-3, caused an increase in the quantity of P protein and an accumulation of protein in neuronal bodies, revealing changes in transport to the neuritic processes. PMID- 19283594 TI - Whole brain size and general mental ability: a review. AB - We review the literature on the relation between whole brain size and general mental ability (GMA) both within and between species. Among humans, in 28 samples using brain imaging techniques, the mean brain size/GMA correlation is 0.40 (N = 1,389; p < 10(-10)); in 59 samples using external head size measures it is 0.20 (N = 63,405; p < 10(-10)). In 6 samples using the method of correlated vectors to distill g, the general factor of mental ability, the mean r is 0.63. We also describe the brain size/GMA correlations with age, socioeconomic position, sex, and ancestral population groups, which also provide information about brain behavior relationships. Finally, we examine brain size and mental ability from an evolutionary and behavior genetic perspective. PMID- 19283596 TI - Comparison of retinal degeneration in the normal goldfish and the megalophthalmic black moor goldfish after optic nerve transection and lens extraction. AB - A comparison in retinal degeneration was studied in the normal goldfish and the megalophthalmic goldfish after optic nerve transection or lens extraction by the TUNEL (Terminal transferase-mediated dUTP nick end labeling) technique. A significant number of TUNEL positive cells appeared in both cases 7 days after injury, with a more prominent result in the megalophthalmic eye. Lens extraction, had less apoptotic cell death in the experimental retinas. PMID- 19283597 TI - Relationship quality and the mentoring of aggressive, high-risk children. AB - We used data from a randomized clinical trial to examine the degree to which relationship quality predicted outcomes for aggressive children in two different mentoring programs. Data were available for 145 aggressive children in Grades 2 and 3. Children were blocked by school and randomly assigned to PrimeTime (n = 75) or Lunch Buddy (n = 70) programs. PrimeTime combined community-based mentoring with child-focused skills training and consultation for parents and teachers, and mentors were extensively trained and supervised. Lunch Buddy was a stand-alone, school-based mentoring program that involved lunchtime visits and a different mentor each semester. PrimeTime children rated their mentors as more supportive than did Lunch Buddy children. Relationship conflict predicted changes in teacher-rated externalizing problems. Ratings of relationship quality interacted with treatment in predicting changes in parent-rated externalizing behavior for PrimeTime children only. PMID- 19283598 TI - Proactive and reactive aggression in a child psychiatric inpatient population. AB - This study examined relations between proactive and reactive aggression and indicators of antisocial behavior (callous/unemotional traits and behavioral consequences) and negative affect (depression and suicidal behavior) in a sample of 105 children admitted to an acute child psychiatric inpatient unit. The majority of the children were male (69%) and ranged from 6 to 12 years of age (M = 9.61, SD = 1.83). Multiple regression analyses revealed that proactive aggression was a unique predictor of antisocial behavior but was not significantly related to negative affect. In contrast, reactive aggression was uniquely positively related to negative affect but unrelated to indicators of antisocial behavior. Findings support unique correlates for proactive and reactive aggression. PMID- 19283599 TI - Enhancing traditional behavioral parent training for single mothers of children with ADHD. AB - Behavioral parent training is an efficacious treatment for attention deficit/hyperactivity disorder (ADHD). However, single-mother households are at high risk for poor outcomes during and following behavioral parent training. This study randomly assigned cohorts of 120 single mothers of children (ages 5-12 years) with ADHD to a waitlist control group, a traditional behavioral parent training program, or an enhanced behavioral parent training program -- the Strategies to Enhance Positive Parenting (STEPP) program. Intent-to-treat analysis demonstrated benefits of participating in behavioral parent training, in general, and the STEPP program more specifically at immediate posttreatment on child and parental functioning. Moreover, the STEPP program resulted in increased engagement to treatment. However, results indicated that behavioral parent training does not normalize behavior for most children and treatment gains are not maintained. PMID- 19283600 TI - Insecure attachment, dysfunctional attitudes, and low self-esteem predicting prospective symptoms of depression and anxiety during adolescence. AB - This study extends the existing adult literature on insecure attachment as a predictor of depression and anxiety by examining these pathways in a sample of adolescents. In addition, dysfunctional attitudes and low self-esteem were tested as mediators of the association between insecure attachment and symptoms of depression and anxiety. Youth (N = 350; 6th-10th graders) completed self-report measures of attachment, dysfunctional attitudes, self-esteem, and symptoms of depression and anxiety in a 4-wave prospective study. Results indicate that anxious and avoidant attachment each predicted changes in both depression and anxiety (after controlling for initial symptom levels). The association between anxious attachment, but not avoidant attachment, and later internalizing symptoms was mediated by dysfunctional attitudes and low self-esteem. Effects remained even after controlling for initial co-occurring symptoms. PMID- 19283601 TI - Parental socialization and adolescents' alcohol use behaviors: predictive disparities in parents' versus adolescents' perceptions of the parenting environment. AB - Among adolescents, many parenting practices have been associated with the initiation and development of drinking behaviors. However, recent studies suggest discrepancies in parents' and adolescents' perceptions of parenting and their links with adolescent use. In this study, we derive two independent sets of underlying parenting profiles (based on parent and adolescent reported behaviors at age 11-12 years), which were then examined in relation to adolescents' drinking behaviors at ages 14 and 17(1/2). Results indicated that the two sets of profiles accounted for little shared variance, with those based on adolescents' reports being stronger predictors of adolescent drinking. Moreover, comparisons of drinking levels across profiles pointed to multiple parenting strategies that may effectively reduce adolescent alcohol experimentation, including simply sustaining a moderate level of awareness of adolescents' whereabouts and activities and avoiding excessive conflict and strictness. PMID- 19283602 TI - Parent-adolescent agreement concerning adolescents' suicidal thoughts and behaviors. AB - Information on history of suicidal thoughts and behaviors is critical in risk assessment, and multi-informant assessment has been recommended. Despite this, relatively little is known about parent-adolescent agreement regarding adolescent suicidality. To examine the extent and predictors of such agreement, 448 psychiatrically hospitalized adolescents and their parents were administered structured interviews assessing suicidal thoughts, plans, and attempts and completed measures of youth internalizing and externalizing behaviors, perceived family social support, and parental distress and psychopathology. Adolescents reported significantly more suicidal ideation, plans, and attempts than parents. Parental history of depression and adolescent perceived family support were associated with significantly greater agreement about suicidality. History of multiple suicide attempts was associated with greater disagreement about suicidality. PMID- 19283603 TI - Child and maternal influence on parenting behavior in clinically anxious children. AB - This study examined child and maternal influences on maternal overinvolvement and negativity. A sample of mothers of children with anxiety disorders (n = 45) and mothers of nonclinical children (n = 46) were observed interacting during a speech preparation task with a child from the same diagnostic group as their child (i.e., anxious or nonanxious) and with a child from a different diagnostic group. During interactions involving children who were not their own, mothers were observed to be more involved with anxious children in comparison to nonclinical children. Mothers of clinically anxious children were observed to be less negative during the interactions with nonclinical children than with clinically anxious children. These results are discussed in relation to the cyclical relationship between parenting behavior and child anxiety. PMID- 19283604 TI - Conners' Teacher Rating Scale for preschool children: a revised, brief, age specific measure. AB - The Conners' Teacher Rating Scale-Revised (CTRS-R) is one of the most commonly used measures of child behavior problems. However, the scale length and the appropriateness of some of the items on the scale may reduce the usefulness of the CTRS-R for use with preschoolers. In this study, a Graded Response Model analysis based on Item Response Theory was applied to the CTRS-R data from 669 preschool children. Children in this data sample ranged in age from 25 to 74 months, and 44.4% were identified as African American, 45.4% Caucasian, 10.2% other. The sample was 53.7% boys and 46.3% girls. A revised version of the scale was developed specifically for use with preschool children. Five items each for the Inattention, Hyperactivity/Impulsivity, and Opposition scales were included in the revised scale. The revised scales significantly reduce the time needed for teachers to complete the measures while retaining the scales' ability to discriminate children with different levels of behavioral problems. PMID- 19283605 TI - Culture, context, and the internalizing distress of Mexican American youth. AB - Latino youth appear to be at higher risk for depression relative to youth from other ethnic groups. This study assessed the relationship between nativity and several forms of internalizing distress among Mexican American middle school students as well as sociocultural factors that may help explain this relationship. Immigrant Mexican American youth (n = 78) reported significantly higher social anxiety and loneliness than U.S.-born Mexican American youth (n = 83). Acculturation stress and English proficiency were identified as significant mediators of these nativity differences. Although internalizing problems and depression symptoms did not vary across nativity groups, both were related to lower affiliative obedience. The findings point to cultural socialization values and contextual influences as important variables in the mental health of youth in immigrant families. PMID- 19283606 TI - Introduction to permutation and resampling-based hypothesis tests. AB - A resampling-based method of inference -- permutation tests -- is often used when distributional assumptions are questionable or unmet. Not only are these methods useful for obvious departures from parametric assumptions (e.g., normality) and small sample sizes, but they are also more robust than their parametric counterparts in the presences of outliers and missing data, problems that are often found in clinical child and adolescent psychology research. These methods are increasingly found in statistical software programs, making their use more feasible. In this article, we use an application-based approach to provide a brief tutorial on permutation testing. We present some historical perspectives, describe how the tests are formulated, and provide examples of common and specific situations under which the methods are most useful. Finally, we demonstrate the utility of these methods to clinical and adolescent psychology by examining four recent articles employing these methods. PMID- 19283607 TI - Prevalence and correlates of drug/alcohol-facilitated and incapacitated sexual assault in a nationally representative sample of adolescent girls. AB - Incapacitated/drug-alcohol facilitated sexual assault (IS/DAFS) is rapidly gaining recognition as a distinct form of assault with unique public health implications. This study reports the prevalence, case characteristics, and associated health risks of IS/DAFS using a large, nationally representative sample of 1,763 adolescent girls. Results indicate that 11.8% of girls experienced at least one form of sexual assault; 2.1% of the total sample experienced IS/DAFS. Thus IS/DAFS accounted for 18% of all reported sexual assaults, with a prevalence of 4.0% among girls 15 to 17 years of age and 0.7% among girls 12 to 14 years of age. Girls with a history of IS/DAFS were significantly more likely than girls with other sexual assault histories to report past-year substance abuse but not significantly more likely than girls with other sexual assault histories to report past-year depression or posttraumatic stress disorder. PMID- 19283608 TI - Mentoring surgeons. AB - From time immemorial mentoring has been the angular stone sustaining the building of medical and surgical education. Good teachers are not necessarily good mentors, and good mentors are not always good teachers. A combination of both is very plausible and should be encouraged. Today, the qualities of a good mentor, in our case the surgeon-mentor, should include respect, time, commitment, trust, determination, encouragement, patience, and opportunity for independence. The mentee would need to respond to similar virtues of trust, encouragement, and respect. The reciprocal consideration of equally divided roles would be clearly desirable. Recognizing the importance of a good mentor and making this role the priority of medical schools would enhance our ability to form better professionals. It would certainly promote professionalism, better patient care, and research. PMID- 19283609 TI - Louis Pasteur surgical revolution. AB - Louis Pasteur (1822-1895) is considered the most notable medical scientist of his time and perhaps one of the most distinguished of all times in the history of medicine. From Dole in France to Paris, from a student of crystals to "living ferments," and from chemistry to biology and medicine, Pasteur changed the world for the benefit of humanity. The genius of Pasteur dealt with the most pressing issues of his time, basing the germ theory on the effects that microorganisms had on fermentation and putrefaction of organic matter, which gave birth to the science of bacteriology. Many other difficult problems in medicine and biology were tackled by Pasteur, culminating in the spectacular results seen with the treatment of rabies. Surgery was no exception to the scientific conquests of Pasteur. The transformation of the surgical world arose from the antiseptic concepts of Lister that were based on the germ theory of the disease, which had been derived from the germ theory of fermentation and putrefaction discovered by Pasteur. The acceptance of these principles represented the surgical revolution brought on by the science of Pasteur, a revolution that is now accepted in our daily care of surgical patients. PMID- 19283610 TI - Ischemic preconditioning protects the pig liver by preserving the mitochondrial structure and downregulating caspase-3 activity. AB - BACKGROUND DATA: The beneficial effects of ischemic preconditioning (IPC) on hepatic ischemia-reperfusion injury (I/RI) have been described. However, the way in which IPC causes the changes in mitochondrial ultrastructure seen in hepatic I/RI is not well understood. OBJECTIVE: The objective of the present study was to determine whether IPC protects the liver from changes in mitochondrial structure and caspase 3 activity in the early phase of post-ischemic injury. METHODS: A pig model consisting of 90 min of hepatic ischemia and 180 min of reperfusion was employed. Eighteen female pigs were randomly divided into three groups: sham operated, non-preconditioned, and ischemic preconditioned (10 min ischemia followed by 10 min reperfusion). Serum concentrations of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and thiobarbituric acid reactive substances (TBARS), as well as bile flow, were measured. Liver biopsies were taken after reperfusion for histological, immunohistochemical (anti-caspase 3), and ultrastructural examinations. RESULTS: The IPC procedure increased bile flow (p < 0.01), reduced serum AST level (p < 0.01), and reduced serum concentration of TBARS at 180 min of reperfusion (p = 0.05). Ischemic preconditioned liver cells had less caspase 3 activity than the non preconditioning group (p < 0.01), and changes in mitochondrial ultrastructure were reduced (p < 0.01). CONCLUSION: IPC exerts a powerful protective effect against hepatic I/RI in the early phase of reperfusion, which may be mediated by preservation of mitochondrial structure and inhibition of caspase-3 activity. PMID- 19283611 TI - Increased expression of connexin43 on the aortic valve in the hypercholesterolemic rabbit model. AB - BACKGROUND: Aortic valve sclerosis is associated with increased risk of cardiovascular death and myocardial infarction. However, the relevance of connexin43 in aortic valve sclerosis remains unclear. We hypothesized that the mechanism regulating aortic valve sclerosis is associated with the alteration of cell-to-cell communication. METHODS: Twenty male New Zealand rabbits were divided into two groups. Group 1 (n = 10) were fed a normal chow diet, while those in group 2 (n = 10) received a diet containing 1% cholesterol for 12 weeks. After utanizing the animals, the aortic valves were excised for analysis. RESULTS: Myofibroblasts and macrophages were more highly expressed in the cholesterol diet group. Osteopontin and connexin43 were found to concentrate within the endothelial layer on the aortic side of the valve leaflets in the cholesterol diet group. A real-time polymerase chain reaction revealed increased connexin43 and osteopontin mRNA levels in the hypercholesterolemic aortic valves. CONCLUSIONS: The present study demonstrates that hypercholesterolemia increases the expression of connexin43 in the rabbit aortic valve. The results suggest that alterations in gap junctional intercellular communication via connexin43 gap junctions may play a role in the development of aortic valve sclerosis. PMID- 19283612 TI - Influence of tumor peroxisome proliferator-activated receptor gamma and delta expression on postoperative mortality of patients undergoing colorectal cancer surgery. AB - PURPOSE: To evaluate the influence of peroxisome proliferator-activated receptor gamma (PPAR gamma) and delta (PPAR delta) expression on postoperative mortality of patients with colorectal cancer (CRC). METHODS: Optimal cutoff values were determined for each relative expression ratio (RER) (RER = PPAR expression of tumor/PPAR expression of normal mucosa) of PPAR, and patients were divided into two groups as follows (PPAR staging): patients with elevated RERs of PPAR gamma (> 2.0) or PPAR delta (> 1.0) were termed Group H, and patients showing none of these elevated RERs of PPARs were termed Group L. Prognostic significance was analyzed by univariate and Kaplan-Meier analyses. RESULTS: In total, 26 CRC patients were studied. Univariate analysis revealed that PPAR gamma (> 2.0/ < or = 2.0) (odds ratio, 11.43; 95% C.I., 1.154-113.1; p = .0373), PPAR delta (> 1.0/ < or = 1.0) (odds ratio, 15.00; 95% C.I., 1.503-149.7; p = .0210) and PPAR staging (H/L) (odds ratio, 63.00; 95% C.I., 4.956-800.8; p = .0014) were significant predictors of postoperative mortality. Kaplan-Meier analysis revealed that the survival curve of patients with CRC was clearly divided by PPAR staging (log rank test, p <.0001). CONCLUSIONS: Evaluation of PPAR gamma and delta expression is useful for predicting postoperative mortality in patients undergoing CRC surgery. PMID- 19283613 TI - The comparison of the effects of different doses of levobupivacaine infiltration on wound healing. AB - INTRODUCTION-AIM: The easiest method in postoperative analgesia is the infiltration of the wound with local anesthetic drugs. Although many local anesthetic drugs have been used for this type of infiltration, studies on levobupivacaine are rare. The aim of this study was to investigate the effects of different concentrations of levobupivacaine infiltration on wound healing. METHOD: Forty female Wistar-Albino rats (280-300 g) were included in the study, which were randomly separated into four groups. Rats were infiltrated with 1.25 mg/mL levobupivacaine in group L(1.25) (n = 10), with 2.50 mg/mL levobupivacaine in group L(2.5) (n = 10), with 3.75 mg/mL levobupivacaine in group L(3.75) (n = 10), and with normal saline in control group (n = 10). Breaking-strength measurements, levels of hydroxyproline, and fibrotic index were evaluated in the tissue samples taken from the rats. RESULTS: When the breaking-strength measurements were evaluated, we have found a significant difference between the control and the study groups (p < 0.05). In the intergroup comparison the difference between groups L(1.25) and L(3.75) was statistically significant (p < 0.05). In all of the levobupivacaine groups the levels of hydroxyproline were higher compared to the control group. Also significant differences were observed between groups L(1.25) and L(2.5) and groups L(1.25) and L(3.75) (p < 0.05). The levels of tissue fibrotic index were higher in all of the levobupivacaine groups compared to the control group (p < 0.05) and also a difference was observed between groups L(1.25) and L(3.75) in terms of tissue fibrotic index (p < 0.05). CONCLUSION: We have concluded that levobupivacaine used in clinical doses have a significant effect on the fastening of wound healing and this effect increases with an increase in the concentration of the levobupivacaine. We believe that levobupivacaine will be more widely preferred in the near future in the postoperative analgesia. PMID- 19283614 TI - The role of suture fixation on mesh contraction after abdominal hernia repair. AB - Mesh contraction is a threat for hernia repair. Our aim is to analyze the effects of fixation techniques on mesh contraction. Forty-eight rats with abdominal wall defects were equally divided into four groups (G): G1 (control) with no repair, G2 defects were repaired with free polypropylene mesh, and G3 and G4 defects were reinforced with prolene mesh fixed by running and interrupted sutures, respectively. The corners of the defect and prolene mesh were marked with silver clips. The contraction rate was calculated by radiological measurement of distances between corner clips and by measuring the mesh areas after harvesting abdominal wall containing the patch. Host reaction was histhopathologically and biochemically examined by inflammation score, fibroblast count, thickness of the granulation tissue, and tissue hydroxyproline level. Distances between corner clips and mesh area have decreased by 31.5% and 26.4% respectively in G2, while in G3 the decrease was 24.4% (p = .008) and 22% (p = .01), respectively. Granulation tissue thickness was highest in the group with mesh fixed by running suture. The tissue hydroxyproline levels were similar in mesh repair groups. Our study suggests that mesh contraction, which reduces mesh surface, occurs during the wound healing process. In order to decrease the contraction rate, it is important to keep the mesh in place until its incorporation into the surrounding tissue. Our results suggest that mesh contraction is minimized by suture fixation, and running fixation suture, which provides more balanced tension around the mesh, seems more beneficial for decreasing contraction rate. PMID- 19283615 TI - Percutaneous intragastric catheter (PIC) for administration of an unpalatable substance to large animals. AB - We studied an easy and reliable technique for administration of an unpalatable substance to large animals. There were three groups of pigs: group I (n = 6) received 1 g ethanol/kg body weight per day orally with water for 24 days, group II (n = 6) received 2 g ethanol/kg orally with water for 24 days and 4 g ethanol/kg via percutaneous intragastric catheter (PIC) for the next 24 days, group III (n = 6) received 6 g ethanol/kg via PIC for 72 days. The catheter was placed after insufflation of the stomach using an orogastric tube. PIC was successfully placed in each pig. No complications occurred during placement. The total amount of the administrated dose was assimilated each time. PIC is a safe, effective, well tolerated, and precise method of administering ethanol that is inexpensive and easy to perform. Ethanol administration via PIC is a convenient and effective mean of exposing animals to high levels of alcohol on a long-term basis. PMID- 19283616 TI - Autogenous greater omentum, as a free nonvascularized graft, enhances bone healing: an experimental nonunion model. AB - Reconstruction of vascularity is an early event in fracture healing and upregulation of angiogenesis may therefore promote the formation of bone. We have investigated the potentiality of autogenous free nonvascularized greater omentum to stimulate the formation of bone in an experimental hypertrophic nonunion model. Twelve dogs assigned into two identical groups underwent a standard nonunion operation. In the experimental group, this was followed by application of autogenous greater omentum as a free nonvascularized graft around the osteotomy gap. Radiographic assessments were conducted time-sequentially until euthanasia 16 weeks after surgery. Histological analysis was performed on the mid radial diaphysis containing the 4-month-old osteotomy site. Radiological and histological properties of the group treated with free transplant of the greater omentum revealed complete union. In contrast, there was no evidence indicating union in the control group. Analyses of the radiological and histological scores confirmed that osteotomies treated with free transplant of the autogenous greater omentum had united, whereas the osteotomies of the control group failed to unite. Significant differences between the mean values for radiological and histological grading score in the control and experimental groups were detected (p < 0.05). We showed that free graft of autogenous greater omentum could stimulate the formation of competent bone in an environment deprived of its normal vascularization. Hence, it could be recommended to enhance healing when the fractures are at risk of nonunion. PMID- 19283617 TI - Suture-free small bowel anastomoses using collagen fleece covered with fibrin glue in pigs. AB - BACKGROUND: Several studies investigating anastomotic healing could objectify that the regularly used suture material leads to an impairment of wound healing due to ischemia at the anastomotic line. This study was initiated to test a hypothesis that a reduction of suture material leading to suture-free glued intestinal anastomoses is feasible and enables an improved wound healing. MATERIALS AND METHODS: Three different types of anastomoses were carried out at the small bowel of 16 pigs. Standard hand-sewn anastomoses, anastomoses with loose-fitting skin staples, and suture-free glued anastomoses using a fibrin covered collagen fleece. When the animals were killed, both gross inspection of the parietes, bursting pressure and tissue for histological study became the basis for evaluation. Analyses were also made regarding the collagen I/III ratio and the expression of MMP 1 and 13. RESULTS: Four leakages at the stapled, one at the sutured, and one at the glued anastomoses occurred. All other anastomoses healed without complications. The bursting pressure did not differ significantly between the groups. The macroscopic inspection and the microscopic examination both showed an improved healing pattern for the material reduced techniques without onset of a deep ulcer at the anastomotic line as seen at the conventional sutured anastomoses. These findings were supported by the immunohistochemical studies. CONCLUSIONS: These observations suggest that a suture-free bowel anastomoses using collagen fleece covered with fibrin glue is technically feasible. Obviously, a reduction of foreign body material at the anastomotic line avoids unnecessary ischemia and thus supports a physiological improved wound healing process. PMID- 19283618 TI - Comparison between gallbladder serosal and mucosal patch in duodenal injuries repair in dogs. AB - BACKGROUND: One of the most challenging problems in clinical surgery is management of an extensive duodenal injury. In its management, there are limitations in using jejunal serosal patch and other conventional methods in specific conditions. This study was performed to compare treatment of large duodenal defects by a gallbladder serosal patch and the gallbladder mucosal patch in a dog as an animal model. METHODS: A duodenal defect (2 cm, about 50% of the total circumference) was created in the second portion of the duodenum in eight dogs. The animals were divided into two equal groups, with group 1 undergoing serosal patch repair and group 2 undergoing mucosal patch repair. The macroscopic and microscopic healing features of the gallbladder serosal and mucosal patch were compared. RESULTS: None of the dogs died due to surgical complications. The whole grafted area was covered by neomucosa at the end of the third week in all animals with the gallbladder serosal patch (group 1). In this group, the scar was small; no significant narrowing of lumen was noted and serosal healing was uniformly complete. In histological examination, a complete coverage of the gallbladder serosal patch by neomucosa consisting of columnar epithelium with short villous formations was observed. In mucosal patch models (group 2), complete epitheliazation, mild fibrosis, and incomplete repair were visible. In histological examination, severe inflammation was noticed too. CONCLUSION: In patients with multiple trauma affecting upper gastrointestinal tracts, use of the gallbladder serosal patch method is easy and reliable. So it may be considered in the surgical management of large duodenal defects, which cannot be repaired by available conventional methods. PMID- 19283619 TI - Principles of anesthesia monitoring--arterial blood pressure. PMID- 19283620 TI - Training health and safety committees to use control banding: lessons learned and opportunities for the United States. AB - Control banding (CB) is a control-focused risk management model that has received international attention. CB strategies are designed to control workplace chemical exposures after the completion of a qualitative risk assessment. Connecticut was one of the first states to provide training on how to use this control-focused tool. Joint labor/management teams and individuals from 34 workplaces attended a control banding workshop and learned how to use one CB model, the United Kingdom (UK) Health and Safety Executive's Control of Substances Hazardous to Health (COSHH) Essentials Toolkit. After the initial training program the investigators used follow-up workshops, questionnaires, site visit data, and case studies to evaluate the training curriculum and assess the utility and effectiveness of this CB strategy. We found that the model is easily learned, although several areas for improvement were identified. Participants from 10 workplaces used COSHH Essentials to evaluate at least one task. The training curriculum was effective in that the agreement between the exposure variables coded by these workplaces and one of the workshop instructors, a certified industrial hygienist (CIH), were highly concordant. The training curriculum and the model promoted a discussion of risk between workers and managers and resulted in the implementation of improvements in the work environment. The model agreed with both the CIH's and the worksites' qualitative risk assessments 65% of the time, and likely over controlled for 71% (5/7) of the cases of nonagreement. Feedback from workshop participants benefits the current dialogue on the implications of implementing CB in the United States. PMID- 19283621 TI - Dust control technology usage patterns in the drywall finishing industry. AB - A telephone survey was conducted to quantify drywall finishing industry usage rates of dust control technology, identify barriers to technology adoption, and explore firm owner perception of risk. Industry use of the following technologies was described: wet methods, respiratory protection, pole sanders, ventilated sanders, and low-dust joint compound. A survey instrument composed of both Likert type scaled items and open-ended items was developed and administered by telephone to the census population of the owners of member firms of trade associations: Finishing Contractors Association and Association of the Wall and Ceiling Industries. Of 857 firms, 264 interviews were completed. Along with descriptive statistics, results were analyzed to examine effects of firm size and union affiliation on responses. Responses to open-ended items were analyzed using content analysis procedures. Firm owners rated the risk of dust to productivity and customer satisfaction as low-moderate. Half rated the dust as having some impact on worker health, with higher impacts indicated by owners of small firms. Among the available control technologies, respiratory protection was used most frequently. Several barriers to implementation of the more effective control technologies were identified. Barriers associated with technology usability, productivity, and cost, as well as misperceptions of risk, should be addressed to improve dust control in the drywall finishing industry. PMID- 19283622 TI - Staff perceptions of syringe dispensing machines in Australia: a pilot study. AB - BACKGROUND/AIMS: Syringe dispensing machines were introduced into needle syringe programs (NSPs) two decades ago. The few published studies on dispensing machines have focused on feedback of machine users and service providers' feedback has rarely been reported. This study obtained the feedback of health staff of NSPs, other sectors of Drug Misuse Treatment Services and of other health services adjoining dispensing machines on the role and effectiveness of dispensing machines. METHODS: Between August and November 2006, questionnaires were anonymously completed by NSP and drug misuser treatment staff in an Area Health Service in Sydney, as well as by the staff of two nondrug-related health services located adjacent to needle syringe dispensing machines. The questionnaire was available in either paper or Internet based forms. RESULTS: Almost 80% of 94 participants rated dispensing machines as either moderately successful or successful in reducing sharing of needles and syringes. Staff considered that introduction of these machines to NSPs had improved services for injecting drug users without increasing unsafe disposal of used equipment, community drug use, or vandalism. However 78% of respondents felt that dispensing machines either reduce or may reduce IDUs' opportunity for staff contact and hence opportunity for engagement or education. The study limitations were noted. CONCLUSION: Syringe dispensing machines are perceived to be a successful and appropriate outlet of NSPs that complement other outlets. Lack of staff-user contact was seen as their main disadvantage. PMID- 19283623 TI - The soundtrack of substance use: music preference and adolescent smoking and drinking. AB - A connection between preferences for heavy metal, rap, reggae, electronic dance music, and substance use has previously been established. However, evidence as to the gender-specific links between substance use and a wider range of music genres in a nationally representative sample of adolescents has to date been missing. In 2003, the Dutch government funded the Dutch National School Survey on Substance Use (DNSSSU), a self-report questionnaire among a representative school-based sample of 7,324 adolescents aged 12 to 16 years, assessed music preference, tobacco, and alcohol use and a set of relevant covariates related to both substance use and music preference. Overall, when all other factors were controlled, punk/hardcore, techno/hardhouse, and reggae were associated with more substance use, while pop and classical music marked less substance use. While prior research showed that liking heavy metal and rap predicts substance use, in this study a preference for rap/hip-hop only indicated elevated smoking among girls, whereas heavy metal was associated with less smoking among boys and less drinking among girls. The types of music that mark increased substance use may vary historically and cross-culturally, but, in general, preferences for nonmainstream music are associated positively with substance use, and preferences for mainstream pop and types of music preferred by adults (classical music) mark less substance use among adolescents. As this is a correlational study no valid conclusions in the direction of causation of the music-substance use link can be drawn. PMID- 19283625 TI - Avian and pandemic influenza: progress and problems with global health governance. AB - Globalization has intensified the health risks posed by pandemic influenza. Effective governance to prepare for, and respond to, a pandemic depends on four key functions: surveillance, protection, response, and communication. Although the global nature of the threat posed is recognized, efforts to strengthen cooperation have only made limited progress. Disease surveillance and communication have benefited from new technologies and harnessing the capacities of both state and non-state actors. The protection and response functions, however, remain focused on domestic populations and are characterized by uncoordinated and, at times, competitive strategies. Global governance of influenza requires strengthening in order to prevent actions that undermine the need to protect the health of all populations. PMID- 19283626 TI - Action on the social determinants of health: a historical perspective. AB - A renewed concern with social factors has emerged in global public health, spearheaded by the World Health Organization's Commission on Social Determinants of Health. The coming decade may see significant health gains for disadvantaged populations if policies tackle the social roots of health inequities. To improve chances of success, global action on social determinants must draw lessons from history. This article reviews milestones in public health action on social determinants over the past 50 years. The goal is to bring into sharper focus the persistent challenges faced by social determinants agendas, along with distinctive opportunities now emerging. The historical record highlights the vulnerability of health policy approaches incorporating social determinants to resistance from entrenched interests. The Commission on Social Determinants of Health can consolidate political support by building collaborative relationships with policymakers in partner countries. However, this strategy must be complemented by engaging civil society constituencies. Historically, successful action on social determinants has been spurred by organized civil society demand. PMID- 19283627 TI - To have and to hold: a cross-cultural inquiry into marital prospects after psychosis. AB - Received wisdom and a substantial body of epidemiological work indicate that early psychosis bodes ill for matrimonial prospects. Using follow-up data from ISoS, the WHO-Collaborative International Study of Schizophrenia, we confirm an earlier local finding that marital success, 15 years after first-break psychosis, is quite favourable in India: 74% for women, 71% for men, compared with elsewhere: 48% for women, 28% for men. This comparative advantage applies to both marriages contracted after onset of psychosis as well as those that survive it, and is the more remarkable for occurring in a culture where the stigma attached to mental illness with regard to marriage is especially heavy. The presence of children and availability of household assistance both appear to enhance odds of successful marriage. That expressed worries about marriage proved so poor a guide to actual performance (and, indeed, survive living proof to the contrary in the families reporting the stigma) suggests that inquiries into stigma should be reworked as larger inquiries into local moral economies of worth. In the dharma governed world of Hindu India resistance to the cultural opprobrium attached to madness is not a strategic assault on a structured source of shame and discrimination, but a tactical manoeuvre in the name of a higher cultural good - family, the lineage and the social order. Restoring this social basis of self respect repairs what would otherwise be a disabling breach in the normal maturation process; developmental continuity, in turn, may help explain India's favourable rates of recovery from psychotic disorder. By the same token, the lack of coordinate processes in cultures where transitions to adulthood are poorly marked and post-hospital expectations are low may help to explain the common experience of 'social defeat', and poor outcome, in the lives of former psychiatric patients in the West. PMID- 19283628 TI - Building peace through participatory health training: a case from Cambodia. AB - This qualitative study examines the significance and effectiveness of participatory health training as a tool for peace building. It does so by analysing a case of training for 'health promoters' run by a Cambodian government health agency. The authors observed participants during the training and interviewed those involved in the courses. A developing capacity for coexistence and reconciliation between individuals who had been on opposite sides during the years of Khmer Rouge terror and continuous internal war was observed among both participants and trainers. Factors embodied in the training that facilitated favourable changes in self and in relations with others were identified as: (1) 'space for dialogue' was created by concrete common public health interests and urgent needs; (2) training took place 'live-in' style in a rural setting; (3) course contents and methods were consistent with peace education; (4) trainers had a conscious function as role models; and (5) there was continuity of effort and consequent accumulation of experience. To build peace, as well as conducting training directly on a technical topic, these essential factors need to be incorporated in the training programmes. PMID- 19283629 TI - The behavioural research agenda in global health: an advocate's legacy. AB - Two of the disciplines that have come to infuse global health with some of its current vibrancy are epidemiology and anthropology, disciplines that focus, in one way or another, on the causal importance of human behaviour in socio political, ecological, evolutionary, and cultural context. One of the little known stories in the history of twentieth century global health involves the works of a number of pioneering interdisciplinary scholar-practitioners, who urged a synthesis of epidemiological and anthropological perspectives in what was then called 'tropical medicine'. One of these pioneers was Frederick L. Dunn, who forwarded lasting insights about the importance of human behavioural research in understanding infectious disease. This article provides a historical-biographical accounting of Dunn's contributions to public health in the second half of the twentieth century, arguing that his persistent advocacy of multi-level, social behavioural research and his notion of 'causal assemblages' were critical in the early development of the twentieth century discipline of global health. PMID- 19283631 TI - The Global Fund 2001-2006: a review of the evidence. AB - The Global Fund to Fight AIDS, Tuberculosis, and Malaria was launched in 2002 with the goals of financing a turnaround in the fight against the three diseases and fundamentally changing the manner in which money is channeled to poor countries. This paper explores the organization's success in fulfilling that mandate over its first five years of operation, examining both its execution of the core business of raising and distributing funds on the basis of proven performance, dramatically increased financing and its realization of this innovative model. The evidence collected demonstrates that, while improvement is needed in a number of areas, the Global Fund has made rapid progress towards realizing its founding ambition and has begun to have its intended impact on global health and development. PMID- 19283632 TI - Harm reduction policies in Argentina: a critical view. AB - The concept of 'harm' underpinning current drug harm reduction policies is most often limited to viral infections and other health consequences for drug users. This paper analyses harm reduction policies in Argentina, with the purpose of challenging and extending this narrow conception of harm to encompass all harms inflicted on drug users, in a context of criminalization of drug use and poverty. Faced with a steep rise in poverty, rapid changes in drug use practices, the quality and prices of drugs, and patterns of morbidity and mortality, Argentina has implemented harm reduction policies specifically for drug users who mostly live in the Greater Buenos Aires impoverished areas. These changes, as well as the Latin American tradition of social and health policies that focus on the collective, subjective, and political-economic aspects of harm, highlight some tensions between the individualistic, public health model structured in the North and its application in Argentina. PMID- 19283633 TI - Maternal deaths in rural Gambia. AB - The objective of this study was to determine causes and contributing factors to maternal deaths in a poor rural setting. We included all maternal deaths (N =42), identified from January to September 2002, in a remote area of The Gambia. To gain a comprehensive picture of medical causes and contributing factors a combination of audit procedure and verbal autopsy was applied. The results showed that anaemia (n =12) was the leading cause of death followed by haemorrhage (n =10), eclampsia (n =8) and obstructed labour (n =8). Placental abruption accounted for 9 of the 10 haemorrhage cases. Substandard obstetric care was identified for the majority of deaths. Substantial inadequacies were revealed at the hospital, characterized by operational difficulties and an uncoordinated emergency preparedness, including malfunction of the blood transfusion service, failure to obtain operative delivery, poor birth monitoring and lack of trained personnel, electricity, medical equipment and drugs. Substandard primary care and logistic difficulties within the referral process further complicated the situation. Delay in seeking care by the cases themselves played a less important role. It was concluded that interventions, addressing the profound deficiencies within the health care system and increasing access to emergency obstetric care, are warranted to reduce maternal deaths in a poor setting such as rural Gambia. PMID- 19283634 TI - 'Halfway people': refugee views of reproductive health services. AB - The objective of this study was to identify factors that facilitate or hinder access to, use of, and satisfaction with reproductive health services in refugee settings, from the perspective of beneficiaries. Rapid appraisal methods included 46 focus group discussions and interviews with over 800 refugees, audits of 14 health facilities, referral hospital reviews, exit interviews with clients, and interviews with health workers. The study was conducted between February and April 2004 in 11 sites in Uganda, Republic of Congo, and Yemen. Reproductive health was clearly on the policy agenda in all countries with stable refugee sites, but problems with implementation and resources were identified. The quality of services was variable, with high staff turnover in some areas affecting relationships with refugee clients. Referral hospitals in host countries were not all equipped to deal with obstetric and other emergencies of either local or refugee populations, including deficiencies in safe blood supplies and antibiotics. Diagnosis and treatment of STIs and HIV/AIDS was frequently inadequate. Gender based violence was the least well addressed aspect of reproductive health. Interest and knowledge about family planning was high, but acceptance was low. It was concluded that progress has been made in reproductive health services for refugees since 1994, however, urgent advocacy and action is required to sustain and improve the situation. Local implementing partners need more support and supervision to develop appropriate service models and to maintain an acceptable standard of care. PMID- 19283635 TI - Novel recruitment strategies to increase participation of women in reproductive health research in India. AB - In India, care seeking for reproductive health among women is inadequate. This poses a unique challenge to researchers recruiting cohorts for studies in clinic based settings. The purpose of this paper is to describe the recruitment process used in a prospective cohort study investigating the relationship between bacterial vaginosis and acquisition of HSV-2 among sexually active women in Mysore, India. Participants were initially recruited from an obstetrics/gynaecology outpatient clinic. Results were compared with a 'community supported' enrolment process, which included community preparation and reproductive health education followed by screening of potential participants. During November 2005, 1,054 women were screened in the clinic. Of the total screened, 246 (23%) were eligible and only 78 (7%) enrolled. Between December 2005 and April 2006, investigators adopted a community supported enrolment process. During that period, 1,077 potential participants were screened, 947 were eligible, and 918 (85%) enrolled. Fifty-six (72%) participants recruited from the clinic returned for their first follow-up visit, compared with 795 (97%) participants recruited using the community supported enrolment process. Since obstetrics/gynaecology departments in India are poor places to recruit non pregnant women of reproductive age, a community supported process yields more eligible potential participants to screen, and results in significantly better study retention. PMID- 19283636 TI - Psychosocial support groups for patients with multidrug-resistant tuberculosis: five years of experience. AB - This detailed case history traces the first 5 years of a psychosocial support group intervention aimed to improve adherence to individualized drug regimens for multidrug-resistant tuberculosis (MDR-TB) in Peru. A total of eight groups were established in metropolitan Lima and two provinces of Peru led by teams of psychiatrists and nurses. The intervention consisted of bi-monthly support groups, recreational excursions, symbolic celebrations, and periodic family workshops. Notably, of the 285 patients who participated in this intervention, only 3.5% defaulted from treatment. Details include the description of services, patient data, major psychosocial difficulties faced by this population, key challenges, and implications. Psychosocial support is a crucial component of treatment for MDR-TB in order to ensure completion of complicated treatment regimens and enable psychosocial rehabilitation after treatment. PMID- 19283642 TI - Psycho-social impact and quality of life of people living with HIV/AIDS in South India. AB - This study examines psycho-social impact of HIV and quality of life of 646 HIV infected persons from a major government sexually transmitted disease (STD) clinic in South India. In this cross-sectional study, data was collected using interview schedule and scales. Nearly 70% had problems in parenting their children after acquiring the infection. Most (88%) of the respondents reported of seeking help from their family members, relatives or close friends at the time of their illness. Among the four categories of stigma, most of them (96%) reported perceived stigma whereas actual stigma was mentioned by only 33%. All four categories of stigma were experienced on a higher proportion by females than males (p<0.05). Each type of stigma was significantly associated with each domain of quality of life of the respondents (p<0.005). Respondents who reported of actual stigma (33%) had significantly good quality of life in their physical domain (49%), psychological domain (48%) and environmental domain (44%). Multivariate analysis showed that gender and marital status had significant association with quality of life. The findings of the study underscore the need for enabling environment through "human force" to uplift their social status and to have a better quality of life. PMID- 19283643 TI - Community factors shaping HIV-related stigma among young people in three African countries. AB - Despite the high prevalence of HIV/AIDS that exists in many sub-Saharan African countries, very little is known of the prevalence and context of HIV-related stigma in these settings. This paper seeks to understand the community-level factors associated with HIV-related stigma among young people in three culturally contrasting African countries: Burkina Faso, Ghana and Zambia. Using nationally representative data on young people (15-24) from Burkina Faso, Ghana and Zambia, the analysis examines the economic, demographic and behavioral dimensions of community environments that shape HIV-related stigma among young people. The results demonstrate a clear influence of the community environment on shaping HIV related stigma among young people. The elements of the community that were significantly associated with HIV-related stigma were the economic and behavioral aspects of the community environment, and there was no evidence of relationships between demographic patterns and HIV-related stigma. Behavioral change interventions must address HIV-related stigma in order to encourage behavior change, and must take into account the social, economic and cultural environments in which young people exist. PMID- 19283644 TI - Emergency department patient perceptions and preferences on opt-in rapid HIV screening program components. AB - The aim of this investigation was to assess emergency department (ED) patients' perceptions and preferences about an opt-in, universal, rapid HIV screening program and identify patient groups who expressed stronger beliefs about components of the testing program. From July 2005 to July 2006, ED patients in the opt-in, universal, rapid HIV screening program were interviewed in person. Multivariable regression models were used to compare participants on their beliefs about the program components. Of the 561 participants, 62.0% had previously been tested for HIV. The majority of participants (58.8%) believed the rapid and standard/conventional HIV tests to be equally accurate, 27.7% believed the rapid test to be less or much less accurate, and 8.7% believed the rapid test to be more or much more accurate. Almost two-thirds (65.1%) favored having a rapid instead of a standard/conventional HIV test, 94.6% wanted the test results within one hour, and 61.3% would be likely or very likely to undergo testing in the ED if it prolonged their ED visit. Almost all (92.5%) believed that their medical care was "not at all" delayed because of being tested, 94.1% believed that testing did "not at all" divert attention from the reason for their ED visit, and 80.9% thought that testing in the ED was "not at all" stressful. In multivariable logistic regression models, males and those with more than 12 years of formal education showed greater concerns about the rapid HIV test's accuracy. Hispanic/Latinos, participants with governmental insurance, and those previously HIV tested were more apt to be screened for HIV even if testing delayed their ED departure. Overall, participants were highly accepting of the components of this opt-in rapid HIV screening program. However, concerns regarding the accuracy of the rapid HIV test might limit test acceptance and should be addressed during pre test information procedures. PMID- 19283645 TI - Burnout during a long-term rehabilitation: comparing low burnout, high burnout - benefited, and high burnout - not benefited trajectories. AB - To focus rehabilitation activities among burnout clients more effectively, it is important to investigate who benefits from burnout interventions. This study (N=85) aimed at identifying burnout trajectories in terms of benefit, that is, subgroups of clients who share similar mean levels and changes in burnout during a one-year rehabilitation intervention (17 days in total) with a six-month follow up. After identifying the burnout trajectories, the relations of the trajectories with factors describing the clients, antecedents, and consequences of burnout during the one-year intervention were examined. Three burnout trajectories were identified by growth mixture modeling: (a) low burnout (n=39), (b) high burnout - benefited (n=29), and (c) high burnout - not benefited (n=17). Positive changes were detected in antecedents and consequences among the clients in the low burnout and high burnout - benefited trajectories. Recovery from burnout was associated with increased job resources and decreased job demands, as well as with increased job satisfaction and decreased depression. It seems that more precise targeting of rehabilitation is needed since the trajectories revealed not only clients with mild symptoms, but also clients who probably received this treatment too late. PMID- 19283646 TI - [Future of continuing medical education in Germany 2009]. PMID- 19283647 TI - [Empirical evidence for the use of anticonvulsants in personality disorders]. AB - OBJECTIVE: There is a common practice of polypharmacy and an increased use of mood stabilizers in personality disorders (PD). This paper reviews all randomized controlled trials (RCTs) of anticonvulsants to evaluate the evidence base supporting their use in treatment of PD. METHODS: German and English language literature cited in Medline and published between 1970 and 2008 was searched using the following terms: Borderline/personality disorder, anticonvulsant, mood stabilizer, carbamazepine, felbamate, gabapentin, lamotrigine, levetiracetam, oxcarbazepine, phenytoine, pregabalin, tiagabine, topiramate, and valproate. RESULTS: Twelve RCTs were identified which included anticonvulsants in treatment of personality disorders. The anticonvulsants valproate and topiramate appeared to have the most empirical support for having a favorable effect on symptoms of borderline personality disorder. Evidence for the use of other anticonvulsants in patients with PD is sparse. CONCLUSIONS: Valproate and topiramate, probably also lamotrigine, carbamazepine, and oxcarbazepine as well, were useful in treating symptoms of affective dysregulation and impulsive aggression in PD. However, further RCTs of anticonvulsants are greatly needed as clinical use of these agents has risen without sufficient evidence supporting their efficacy and safety in personality disorders. PMID- 19283648 TI - [Psychoeducation for in-patient anxiety--the evaluation of the cognitive behavioral group programme "PAsta"]. AB - BACKGROUND: The object of this study is to develop and evaluate a psychoeducational programme that has been specifically designed for in-patients who suffer from heterogenous anxiety disorders. METHOD: 28 in-patients participated in psychoeducational group sessions in addition to psychiatric treatment as usual (TAU). This group was compared to a group of 13 patients who had received TAU without the group sessions. Using a pre-post design, the two groups were compared in terms of illness knowledge, mental health and self efficacy. RESULTS: Analysis of the results showed a significant increase in knowledge of anxiety among participants of the psychoeducational group (d = 0.80). The effect sizes from 0.12 - 0.60 indicate clinical relevant improvement in depressive and anxiety symptoms, and a tendency towards improved self-efficacy (d = 0.20). CONCLUSIONS: The main objective of the group programme was achieved; that is, it increased illness knowledge. Despite the short measurement period, the importance of the psychoeducational group programme in the treatment of acute in-patients was confirmed. PMID- 19283649 TI - [Physical activity and prevention of Alzheimer's dementia: current evidence and feasibility of an interventional trial]. AB - There is accumulating evidence from animal and epidemiologic studies that physical exercise is neuroprotective in healthy animals and humans and can prevent cognitive decline in chronic neurodegenerative processes like Alzheimer's dementia. However, data from well-designed interventional, randomized non pharmacologic trials is lacking in contrast to other areas of medicine like prevention of hypertension, diabetes or the antipsychotic-associated metabolic syndrome. The demonstration of a potential positive effect of physical exercise on preventing dementia using a controlled study design would represent a significant progress in the prevention of dementia and public health, especially as long as other treatments for dementia prevention are lacking. PMID- 19283650 TI - [Psychiatric disorders and childhood trauma in prisoners with antisocial personality disorder]. AB - Previous studies indicate high prevalence rates of mental disorders and trauma among prisoners. Based on a sample of 102 male German prisoners, the comorbidity and childhood trauma experiences in 72 criminals with antisocial personality disorder were investigated. Furthermore, associations of antisocial personality disorder and early traumatic experiences with the age at first conviction and the lifetime months of imprisonment were examined. Subjects had high rates of comorbid lifetime and current disorders as well as childhood trauma experiences. Physical abuse in childhood and adolescence was identified as a predictor for lifetime months of imprisonment, antisocial personality disorder was found to be a predictor for the age at first conviction. Our findings confirm the hypothesis of prisoners with antisocial personality disorder being a severely traumatized population with serious mental disorders. Traumatic childhood experiences and antisocial personality disorder are associated with criminality variables. This has important implications on preventive treatments as well as on how prison services are addressing these problems. PMID- 19283651 TI - [Holderlin--or the question of the meaning of psychosis]. AB - Friedrich Holderlin (1770-1843) is one of the most important German poets. Actual research into his life and work has shown new aspects in his thinking concerning questions about subjectivity, sense of life and psychosis. We follow these lines using a hermeneutical method. In his late poems the experience of schizophrenic alienation appears metaphorically speaking like an ebbing of a former plenitude of meanings or as if he were decentered from his own life. Holderlin names it an "uninvolved" view onto the ordinary life. Holderlin invites and enables us via his offer for an innerperspective understanding of the schizophrenic experience of alienation to deal fairly and respectfully with schizophrenic patients as if we were "alienists" (E. Straus). PMID- 19283652 TI - [Sporadic adult form of nemaline myopathy--a difficult differential diagnosis]. AB - We report on a 79 years old female patient with slowly progressive painful weakness of the proximal upper and lower limbs. CK was slightly elevated (242 U 7 l). MRI guided biopsy of the proximal lower limbs revealed the rare findings of nemaline myopathy. PMID- 19283653 TI - [Frontotemporal lobar degeneration. Part 1: diagnosis and treatment]. PMID- 19283654 TI - Analysis of murine vascular function in vivo by optical coherence tomography in response to high-fat diet. AB - In this study, we demonstrate the application of optical coherence tomography (OCT) as a contactless imaging technique to analyze vasodynamics in small blood vessels in vivo. The transluminal OCT imaging of vessels avoids micro traumata in the endothelium and circumvents surgical intervention. It can be performed in the intact perfused vessel and provides a new method to measure vascular function and dynamics in vivo. The resolution of 10 mum and the velocity of image acquisition are adequate to detect differences in the inner diameter, the maximal velocity, or the time to half-maximal diameter change of small vessels. We applied this new technology to study the vascular dynamics in small vessels of 6- and 20-week-old C57BL/6 mice in vivo. In addition, we determined by this technique the impact of a high-fat diet for 14 weeks on vascular function in 20-week-old animals. The diameter of the saphenous artery was increased under resting conditions, after vasoconstriction and after vasodilatation in 20-week-old animals on normal chow and high-fat diet, compared to 6-week-old animals. High-fat diet caused a significantly impaired vasoconstriction in the saphenous artery. The maximal velocity of diameter changes of the saphenous artery was determined by time resolved OCT imaging. A significant reduction of this parameter was found during vasoconstriction in 20-week-old mice on high-fat diet, compared to 6-week-old animals. In conclusion, transluminal optical coherence tomography imaging is a novel and useful technique to analyze the impaired vasodynamics of small arteries in response to high-fat diet in vivo. PMID- 19283655 TI - The efficacy of Quintero staging system to assess severity of twin-twin transfusion syndrome treated with laser therapy: a systematic review with meta analysis. AB - Severity of twin-twin transfusion syndrome (TTTS) is classified in five stages according to Quintero staging. However, the efficacy of such staging was recently debated. We reviewed the efficacy of Quintero staging to predict survival rate in TTTS treated with laser therapy. Articles reporting survival rate for each stage in TTTS treated with laser therapy were reviewed. Number of twins alive per pregnancy (NAP) was compared between early (I + II) and advanced (III + IV) stages and within stages. Meta-analysis was performed according to Meta-analysis Of Observational Studies in Epidemiology guidelines. Heterogeneity was tested with chi-square for heterogeneity at a significance level of P < 0.10, and random or fixed models were generated as appropriate. A P value < 0.05 was considered statistically significant. NAP was similar between early (zero survivors: 34/228, 15%; one survivor: 49/228, 21%; two survivors: 145/228, 63%) and advanced stages (zero survivors: 38/214, 18%; one survivor: 64/214, 30%; two survivors: 112/214, 52%; P > 0.05) except for one survivor ( P < 0.05). A trend for increased NAP was observed in all stages. Because clinically relevant differences were not observed, laser therapy is the optimal treatment for all stages. As Quintero staging does not provide information about prognosis, a new staging system is proposed. PMID- 19283656 TI - Neonate exposure to thimerosal mercury from hepatitis B vaccines. AB - Infant exposure to ethylmercury (EtHg) has not only increased but is starting earlier as a result of the current immunization schedule that uses thimerosal containing vaccines (TCVs). Although vaccination schedule varies considerably between countries, infants in less-developed countries continue to be exposed to EtHg derived from more affordable TCVs. We studied the exposure of newborns to EtHg from hepatitis B vaccines; hospital records (21,685) were summarized for the years 2001 to 2005 regarding date of birth, vaccination date, and birth weight. Most of the vaccinations occurred in the first 24 hours postdelivery; over the 5 years, there was an increase in vaccinations within hours of birth (same day), from 7.4% (2001) to 87.8% (2005). Nearly 94.6% of infants are now being vaccinated within the first 24 hours. Range of mercury exposure spread from 4.2 to 21.1 microg mercury/kg body weight for those receiving TCVs with the highest thimerosal concentration; these exposure levels are conservative for 2% of children receiving vaccines within 2 to 3 postnatal days, when they are still going through physiological postnatal weight loss. Because of the particular timing (transitioning from in utero to ex utero metabolism) and specific aspects of exposure (i.e., parenteral mode, bypassing gastroenteric barriers) and dose (related to vaccine manufacturer and with variation in birth weight), this study reveals critical issues that can modulate toxicokinetics and toxicodynamics of organomercurials in neonates. PMID- 19283657 TI - [Injury of ICA during endonasal sinus surgery and management by endovascular stent application]. AB - BACKGROUND: Injury of the internal carotid artery (ICA) during endonasal sinus surgery is rare but a life-threatening situation for the patient. For adequate treatment endovascular therapy as embolization or balloon occlusion is usually performed although there is a high risk of irreversible neurological deficits. CASE REPORT: We report on a twenty-five-year-old female patient with acute exacerbated chronic polyposis nasi suffering from an injury of the ICA during endonasal sinus surgery. After intraoperative identification of the site of hemorrhage the massive bleeding could temporarily be controlled by a nasal pack. The woman was immediately transferred to a neuroradiological department. After angiographic localization of the vessel defect the bleeding could be stopped by application of a vascular stent without any neurological deficits during or after the procedure. Anatomical characteristics of the ICA in the sphenoid sinus, and recommended emergency treatment and preventive measures are discussed. CONCLUSION: Although vascular stents are difficult to place in the intrasphenoidal course of the ICA, it provides an effective closure of the vascular injury and above all a sufficient cerebral perfusion, therefore, the risk of neurological defects can be reduced. PMID- 19283658 TI - On "Analysis of flow changes to the foot after sacrifice of one of the major arteries (J Reconstr Microsurg 2009;25:35-39)". PMID- 19283659 TI - Effect of IIb-IIIa glycoprotein inhibitors in a partial limb amputation model submitted to warm ischemia in rats. AB - Viability and functional results of a segment replantation depend on the prevention of deleterious effects of ischemia. Prolonged ischemia leads to alterations in the microcirculation: thrombosis, edema, production of oxygen free radicals, and platelet aggregation. The effect of IIb-IIIa glycoprotein inhibitors was tested in a partial limb amputation model submitted to warm ischemia. The male Wistar rats were divided into four groups: G1 with 0 hours of ischemia and saline ( N = 20), G2 with 6 hours of ischemia and saline ( N = 24), G3 with 6 hours of ischemia and abciximab ( N = 23), and G4 with 6 hours of ischemia and tirofiban ( N = 29). The limbs were observed for 7 days and classified as viable or nonviable. Viability and mortality rates were obtained and analyzed by Q-square and Fisher exact tests ( P < 0.05). The viability rates were 100% (G1), 30% (G2), 77.78% (G3), and 80.95% (G4). G2 was statistically different from G1, G3, and G4. G1, G3, and G4 were not statistically different. Transoperative and postoperative mortalities were not statistically different. The administration of abciximab and tirofiban improved limb salvage after ischemia and reperfusion and did not modify mortality rates significantly. PMID- 19283660 TI - [Imported suicide rates]. PMID- 19283661 TI - Diabetes enhances the expression of H-ras and suppresses the expression of EGFR leading to increased cell proliferation. AB - EGFR kinase activity triggers numerous signaling pathways, such as the Ras/Raf/MAPK cascade, leading to the activation of various mitogen activated protein kinases, which are implicated in cell proliferation through induction of several genes, including c-fos. The possible effect of diabetes on the expression of the oncogenes EGFR, H-ras and c-fos was investigated in an experimental model of chemically induced oral oncogenesis in normal and diabetic (type I) Sprague Dawley rats. Thirteen diabetic and twelve normal rats developed cancer after 4NQO treatment, while six diabetic and six normal animals were used as controls. The biopsies were classified pathologically (ranging from dysplasia to moderately differentiated oral squamous cell carcinoma) and were studied immunohistochemically. Several representative histological regions from each biopsy were analysed in regard to EGFR, H-ras and c-fos expression, and a comparison between normal and diabetic rats was effected. A trend of decreased EGFR expression in diabetic compared to normal rats was revealed throughout oncogenesis, which was significant in the stage of dysplasia (P<0.05). On the contrary, a trend of increased H-ras expression was observed in diabetic compared to normal rats during oncogenesis, which rose significantly in early invasion and well differentiated OSCC (P<0.001 and P<0.01 respectively). Finally, no statistical differences concerning c-fos expression were detected between diabetic and normal animals. In conclusion, it seems that diabetes reduces the expression of EGFR and initiates the Ras/Raf/MAPK signal transduction pathway by enhancing activation of other signalling molecules, such as the diabetes associated Insulin Receptor Substrate-1, leading to increased cell proliferation without c-fos involvement. PMID- 19283662 TI - Ultrastructure of myotendinous junctions in tendon-skeletal muscle constructs engineered in vitro. AB - During development, the interaction between tenocytes and myotubes leads to the formation of highly specialized muscle-tendon structural interfaces: myotendinous junctions (MTJs). Structural integrity of MTJs is critical for force transmission from contracting muscle through tendon to bone. We recently developed an in vitro model of three-dimensional (3-D) skeletal muscle-tendon constructs to address mechanisms of the MTJs development. We hypothesized that engineered in vitro 3-D skeletal muscle-tendon constructs would develop MTJs ultrastructurally resembling those found during fetal development in vivo. To test this hypothesis we compared MTJs structures in vivo to those developed in 3-D skeletal muscle constructs co cultured with engineered self-organized tendon constructs (SOT), or segments of adult (ART) or fetal rat tail (FRT) by means of electron microscopy. Our study showed that at sites of termination some of the myofibers of the engineered 3-D skeletal muscle-FRT and -SOT constructs displayed emerging finger-like sarcolemmal projections surrounded by collagen fibers. These structures resemble fetal MTJs in vivo. Muscle-ART constructs did not develop MTJs. Muscle-FRT constructs in addition to muscle and tendon also demonstrated well developed cartilage, possessing high potential for development into bone. The muscle-FRT construct model could be used for studies of developmental mechanisms involved in the establishment of interfaces among all four muscular-skeletal tissues: muscle, tendon and cartilage/bone. PMID- 19283663 TI - The role of vascular adhesion molecules PECAM-1 (CD 31), VCAM-1 (CD 106), E selectin (CD62E) and P-selectin (CD62P) in severe porcine pancreatitis. AB - Inflammatory cytokines have been shown to mediate organ damage by their action on vascular endothelia and leukocytes, in part by upregulating the expression of adhesion molecules, which in turn convey transmigration of leukocytes into tissue. The upregulation and activation of vascular cell adhesion molecules on the endothelial cells avail firm leukocyte adhesion to the vascular endothelium and enhance their transmigration and consecutive tissue injury. The aim of this study was to evaluate the expression of vascular adhesion molecules CD 31 (PECAM 1), CD 106 (VCAM-1), CD 62E (E-Selectin) and CD 62P (P-Selectin) in the pancreas and distant organs of pigs suffering from acute necrotizing pancreatitis (AP). AP was induced in 13 pigs by a combination of intravenous cerulein and intraductal glycodeoxycholic acid. For immunostaining of vascular adhesion molecules slides of porcine pancreas, lung, kidney and liver tissue were stained with monoclonal antibodies (Ab) against PECAM-1-1, VCAM-1 E- and P- SELECTIN. The endothelial cell expression of CD 31 (PECAM-1), CD 106 (VCAM), CD 62E (E-Selectin) and CD 62P (P-SELECTIN) in severe porcine pancreatitis is detectable and upregulation is partly significantly. PMID- 19283664 TI - Comparison of histopathological features of Vibrio cholerae O1 El Tor and O139 Bengal infections in rabbit intestinal mucosa. AB - Vibrio cholerae is the causative agent of the infectious disease, cholera. The bacteria adhere to the mucosal membrane and release cholera toxin, leading to watery diarrhea. There are >100 serovars of V. cholerae, but the O1 and O139 serovars are the main causative agents of cholera. The present study aimed to compare the severity of intestinal mucosal infection caused by O1 El Tor and O139 V. cholerae in a rabbit ileal loop model. The results showed that although the fluid accumulation was similar in the loops inoculated with O1 and O139 V. cholerae, the presence of blood was detected only in the loops inoculated with the O139 serovar. Serosal hemorrhage was confirmed by histopathological examination and the loops inoculated with O139 showed massive destruction of villi and loss of intestinal glands. The submucosa and muscularis mucosa of the ileum showed the presence of edema with congested blood vessels, while severe hemorrhage was seen in the muscularis propria layer. The loops inoculated with O1 El Tor showed only minimal damage, with intact intestinal villi and glands. Diffuse colonies of the O139 serovar were seen to have infiltrated deep into the submucosal layer of the intestine. Although the infection caused by the O1 serovar was focal and invasive, it was more superficial than that due to O139, and involved only the villi. These observations were confirmed by immunostaining with O1 and O139 V. cholerae-specific monoclonal antibodies. The peroxidase reaction demonstrated involvement of tissues down to the submucosal layer in O139 V. cholerae infection, while in O1 El Tor infection, the reaction was confined mainly to the villi, and was greatly reduced in the submucosal region. This is the first reported study to clearly demonstrate the histopathological differences between infections caused by the O139 Bengal and O1 El Tor pathogenic serovars of V. cholerae. PMID- 19283665 TI - Golgi apparatus localization of ZNT7 in the mouse cerebellum. AB - We have recently reported that four members of the zinc transporter (ZNT) family, ZNT1, ZNT3, ZNT4, and ZNT6, are abundantly expressed in the mouse cerebellum. In the present study, we reported that ZNT7 was present throughout the cerebellar cortex. ZNT7 immunoreactivity was predominately present in the somas and primary dendrites of the Purkinje cells. ZNT7 was also present in the Bergmann glial cell bodies as well as their radial processes, which extended into the molecular cell layer. Confocal immunofluorescence results demonstrated that the expression of ZNT7 overlapped with that of TGN38 in the somas of the Purkinje cells and granule cells. Immuno-electron microscopic study showed that ZNT7 was localized to the membrane of the Golgi apparatus in the somas of the Purkinje cells, Bergmann glial cells, and granule cells. Western blot analysis demonstrated that a considerable amount of ZNT7 was expressed in the cerebellum. These findings suggest a significant role of ZNT7 in zinc homeostasis in the mouse cerebellum. PMID- 19283666 TI - Metallic gold treatment reduces proliferation of inflammatory cells, increases expression of VEGF and FGF, and stimulates cell proliferation in the subventricular zone following experimental traumatic brain injury. AB - Traumatic brain injury represents a leading cause of morbidity in young individuals and there is an imperative need for neuroprotective treatments limiting the neurologic impairment following such injury. It has recently been demonstrated that bio-liberated gold ions liberated from small metallic gold implants reduce inflammation and neuronal apoptosis, while generating an increased neuronal stem cell response following focal brain damage. In this study mice were subjected to a unilateral traumatic cryo-lesion with concomitant injection of 25-45 microm gold particles near the lesion. Placebo-treated mice subjected to cryo-lesion served as controls. The effects of gold-treatment were investigated by examining gold-induced growth factor expression (VEGF and FGF) in the first two weeks after the insult, and the extent of the neurostimulatory effect of gold was explored by comparing cell proliferation in the subventricular zone as judged by immunohistochemical staining for CDC47. Vimentin staining revealed a decrease in activated microglia and a transient astrogliosis in response to the gold liberation. Moreover, gold ions significantly increase the expression of VEGF and FGF following trauma and a significant increase in cell proliferation in both the ipsilateral and the contralateral subventricular zone was found in response to gold-treatment. IN CONCLUSION: we confirmed the previously demonstrated anti-inflammatory effect of bio-liberated gold ions, and further show that metallic gold increases growth factor expression and adult neurogenesis. PMID- 19283667 TI - Cardiac ischemia and reperfusion in spontaneously diabetic rats with and without application of EGb 761: II. Interstitium and microvasculature. AB - Besides alterations in cardiomyocytes themselves, diabetic cardiopathy is characterized by interstitial and microvascular disorders. On the assumption that a specific heart muscle disease develops due to permanently increased oxidative stress on liberation of oxygen-free radicals, adjuvant application of antioxidative therapeutics appears promising in preventing or delaying long-term diabetic complications and protecting the myocardium against acute ischemia. We have investigated the effects of Ginkgo biloba extract (EGb 761), a radical scavenger, against diabetes-induced myocardial interstitium and microvasculature damage, and against additional ischemia/reperfusion injury in spontaneously diabetic BioBreeding/Ottawa Karlsburg (BB/OK) rats modelling diabetic cardiac infarction. Morphological and morphometric parameters in the heart muscle were evaluated by light and electron microscope. We used immunohistochemistry to investigate collagen protein expression as a marker for tissue remodelling together with endothelial nitric oxide synthase (eNOS) protein expression as a marker for endothelial-dependent vasodilation. We also evaluated inflammation response caused by neuropeptide Substance P and interacting mast cells in the diabetic heart. Our results revealed that A) Diabetic myocardium appears more vulnerable to ischemia/reperfusion injury than normal myocardium with regard to myocardial interstitium and microvessel ultrastructure, as well as eNOS protein expression; B) Inflammation response increases in diabetic animals exposed to ischemia/reperfusion injury compared to controls; C) Pre-treatment of diabetic myocardium with EGb results in an improvement of impaired endothelial-dependent vasodilation in diabetes and additional ischemia/ reperfusion, diminished mast cell and substance P accumulation, and better preserved myocardial ultrastructure compared to unprotected myocardium. In conclusion, EGb may act as a potent therapeutic adjuvant in diabetics with respect to ischemic myocardial injury, and may contribute to preventing late complications in diabetic cardiopathy. PMID- 19283668 TI - Cytochemical and biochemical evidences for a complex tridimensional structure of the hamster zona pellucida. AB - Zona pellucida (ZP) is an extracellular matrix that surrounds eggs and pre implantation embryos and is required for in vivo fertility. A key event in successful fertilization is sperm binding to the surface of the ZP. It has been previously described that the hamster sperm binds mainly the outer region of the ZP which corresponds to the porous region in contact with the cumulus cells. Using ultrastructural cytochemistry approaches with an antibody developed against porcine ZP, this study shows that the pig ZP shares epitopes with some rodent species like hamster, rat and mouse. In the hamster, these epitopes are located mainly in the outer region of the ZP of preovulatory and ovulated oocytes. By means of biochemical approaches it was demonstrated that 1) the antibody is specific for the native hamster ZP3, 2) four different bands with a molecular weight of 67, 60, 48 and 38 kDa after N-linked deglycosylation suggesting that the hamster ZP is formed by four proteins, and 3) the different composition observed in the outer region of the hamster ZP could be due to a specific supramolecular structure that makes some epitopes accessible for the antibodies. In summary, this study provides evidence that the different composition observed in the different regions of the ZP is mediated by a different organization of the components of the ZP produced during the oocyte maturation. This different organization could be responsible for the different sperm binding affinity observed for sperm to the outer region versus the inner region of the ZP. PMID- 19283669 TI - Skeletal muscle fibre plasticity in response to selected environmental and physiological stimuli. AB - Skeletal muscle constitutes a highly adaptable and malleable tissue that responds to environmental and physiological challenges by changing its phenotype in terms of size and composition, outcomes that are brought about by changes in gene expression, biochemical and metabolic properties. Both the short- and long-term effects of nutritional alterations on skeletal muscle homeostasis have been defined as the object of intensive research over the last thirty years. This review focuses predominantly on assimilating our understanding of the changes in muscle fibre phenotype and functional properties induced by either food restriction or alternatively existing on a high fat diet. Firstly, food restriction has been shown in a number of studies to decrease the myofibre cross sectional area and consistently, it has been found that glycolytic type IIB fibres are more prone to atrophy than oxidative fibres. Secondly, in rodents, a high fat diet has been shown to induce an oxidative profile in skeletal muscle, although obese humans usually show higher numbers of glycolytic type IIB fibres. Moreover, attention is paid to the effect of prenatal maternal food restriction on muscle development of the offspring in various species. A key point related to these experiments is the timing of food restriction for the mother. Furthermore, we explore extensively the seemingly species-specific response to maternal malnutrition. Finally, key signalling molecules that play a pivotal role in energy metabolism, fibre type transitions and muscle hypertrophy are discussed in detail. PMID- 19283670 TI - Induction system of neural and muscle lineage cells from bone marrow stromal cells; a new strategy for tissue reconstruction in degenerative diseases. AB - Since bone marrow stromal cells (MSCs) are easily accessible both from healthy donors and patients, and can be expanded on a therapeutic scale, they have attracted attention for cell-based therapy. MSCs contribute to the protection of host tissue after transplantation by Immune modulation and trophic effect. They also have an ability to differentiate into other cell kinds that will replenish lost cells in the degenerated tissue. This review discusses the potential of MSCs for tissue reconstruction in neuro- and muscle-degenerative diseases and their differentiation capacity into functional cells. PMID- 19283671 TI - c-KIT signaling as the driving oncogenic event in sub-groups of melanomas. AB - As we enter the era of targeted therapy for melanoma, attempts are being made to sub-group tumors on the basis of their driving oncogenic mutations, with the hope of developing truly personalized therapeutic regimens. c-KIT is a receptor tyrosine kinase whose aberrant activation is implicated in the progression of gastrointestinal stromal tumors and some acute myeloid leukemias. The role of c KIT signaling in melanoma has been controversial; although c-KIT activity is critical to melanocyte development, its expression tends to be lost in most melanomas. Some reports have even shown that the re-expression of c-KIT induces apoptosis in melanoma cell lines. The recent publication of work showing the presence of activating c-KIT mutations in acral and mucosal melanomas, as well as melanomas arising on skin with chronic sun damage, has renewed interest in c-KIT signaling in melanoma. Recent work from our own laboratory has further identified melanomas with constitutive c-KIT signaling activity resulting from c-KIT receptor overexpression. Although the initial clinical trials of the c-KIT inhibitor imatinib mesylate in melanoma were negative, some dramatic responses have been seen in patients with very high c-KIT expression and/or documented activating mutations, fostering the belief that focused studies in patients selected on the basis of c-KIT mutational status will yield more encouraging results. The current review discusses the role of c-KIT signaling in melanoma progression and how this new information can be applied to the targeted therapy of melanoma. PMID- 19283672 TI - Lysyl oxidases in mammalian development and certain pathological conditions. AB - Lysyl oxidase (LOX) catalyzes the oxidation of the side chain of a peptidyl lysine converting specific lysine and hydroxylysine residues of alpha-aminoadipic delta-semialdehydes, which form covalent crosslinks in collagens and elastin. Five different but closely related lysyl oxidase isoenzymes have been identified to date, and they seem to have overlapping functions in many tissues. Modification of the extracellular matrix by lysyl oxidases has been shown to be a critical contributor to the development of various organs and certain pathological conditions. PMID- 19283673 TI - Modified Gleason grading. An updated review. AB - At an ISUP (International Society of Urological Pathology) consensus conference in 2005 in San Antonio, Texas, the old Gleason grading system for prostatic carcinoma from 1966 underwent its first major revision. With this modified Grading system a shift of the most frequent Gleason scores from 6 to 7a (3+4) in biopsy specimens and an increased degree of agreement between specimens of biopsies and radical prostatectomies with carcinoma of the prostate could be demonstrated. After modified grading of GS 3+4=7a tumours 95% were stage pT2, while 79% of GS 4+3=7b tumours were stage pT3-4. In cases with PSA <10 ng/ml and tumour extent <20% the most frequent Gleason scores were 6 and 7a. Cases with serum PSA >10 ng/ml or tumour extent >20% had higher scores (7b or higher). Cancers with tumour infiltration of <1mm in one of 12 cores and PSA <10 ng/ml were mainly low grade (Gleason scores 6 and 7a) and may correspond to so called insignificant carcinoma of the prostate. CONCLUSION: With the modified Gleason system, grade, stage, tumour extent and serum PSA show good correlations and characterize the difference between low and high grade malignancy of prostate carcinoma. PMID- 19283674 TI - Evaluation of cancer records from 2000-2004 in Denizli, Turkey. AB - Objective information about cancer incidence is important for planning control programs. We examined the distribution of cancer cases recorded in Denizli province, Turkey. A total of 2185 cancer cases reported to the Denizli Province Health Ministry's Cancer Early Diagnosis Center during the years 2000-2004 were evaluated for sociodemographic characteristics, cigarette use, family history, and organ systems. Among these cases, 56% were male and 44% were female; 45.1% of the patients had smoked cigarettes at some time and there was a 10-fold increase in lung cancer and a 4-fold increase in urinary cancers among cigarette smokers (P < 0.001). We found that 34.4% of the cancer cases were diagnosed as localized, 27.9% had a more extensive distribution and 21.8% were in metastasis. The most frequent types were urinary cancers at 26.4%, gastrointestinal cancers at 19.2% and respiratory cancers at 18.9%; there was a significant increase in gastrointestinal, blood and skin cancers over the years. Lung (14.9%), breast (14.1%), bladder (8.0%), prostate (5.3%), and lymphatic (4.8%) cancer cases were the most common. PMID- 19283675 TI - Imprinted gene expression in in vivo- and in vitro-produced bovine embryos and chorio-allantoic membranes. AB - Cloning by nuclear transfer is often associated with poor results due to abnormal nuclear reprogramming of somatic donor cells and altered gene expression patterns. We investigated the expression patterns of imprinted genes IGF2 and IGF2R in 33- to 36-day bovine embryos and chorio-allantoic membranes derived from in vivo- and in vitro-produced embryos by somatic cell nuclear transfer (SCNT), parthenogenetic activation, and in vitro fertilization (IVF). There was a lower IGF2 expression rate in the SCNT (0.19) and parthenogenetic (0.02) groups when compared to in vivo and IVF embryos (2.01; P < 0.05). In the chorio-allantoic membranes, IGF2 showed a baseline expression pattern (P < 0.05) in parthenotes (0.001) when compared to in vivo, IVF (3.13), and SCNT (0.98) groups. IGF2R was less expressed (P < 0.05) in SCNT chorio-allantoic membranes (0.25) when compared to the in vivo group. The low expression of IGF2 in parthenogenetic embryos and chorio-allantoic membranes confirms its imprinted status in cattle. Alterations in the relative frequency of IGF2 and IGF2R transcripts were observed in SCNT derived bovine embryos and chorio-allantoic membranes, respectively, supporting the hypothesis that abnormalities in the expression of imprinted genes are causes of the low efficiency of SCNT procedures in this species. PMID- 19283676 TI - An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees. AB - Traditionally, molecular studies of plant species have used leaves as the source of DNA. However, sampling leaves from tall tree species can be quite difficult and expensive. We developed a sequence of procedures for using stem bark as a source of DNA from Leguminosae trees of the Atlantic Forest and the Cerrado. Leguminosae is an important species-rich family in these two highly diverse and endangered biomes. A modified CTAB protocol for DNA isolation is described, and details of the procedures for sampling and storage of the bark are given. The procedures were initially developed for three species, and then their applicability for 15 other species was evaluated. DNA of satisfactory quality was obtained from the bark of all species. The amounts of DNA obtained from leaves were slightly higher than from bark samples, while its purity was the same. Storing the bark frozen or by drying in silica gel yielded similar results. Polymerase chain reaction amplification worked for both plastid and nuclear genomes. This alternative for isolating DNA from bark samples of trees facilitates field work with these tree species. PMID- 19283677 TI - Estimates of genetic trend for carcass traits in a commercial broiler line. AB - Data from the slaughter of 24,001 chickens that were part of a selection program for the production of commercial broilers were used to estimate genetic trend for absolute carcass (CW), breast meat (BRW), and leg (LW) weights, and relative carcass (CY), breast meat (BRY), and leg (LY) weights. The components of (co)variance and breeding values of individuals were obtained by the restricted maximum likelihood method applied to animal models. The relationship matrix was composed of 132,442 birds. The models included as random effects, maternal additive genetic and permanent environmental for CW, BRW, LW, CY, and BRY, and only maternal permanent environmental for LY, besides the direct additive genetic and residual effects, and as fixed effects, hatch week, parents' mating group and sex. The estimates of genetic trend were obtained by average regression of breeding value on generation, and the average genetic trend was estimated by regression coefficients. The genetic trends for CW (+6.0336 g/generation), BRW (+3.6723 g/generation), LW (+1.5846 g/generation), CY (+0.1195%/generation), and BRY (+0.1388%/generation) were positive, and they were in accordance with the objectives of the selection program for these traits. The genetic trend for LY ( 0.0019%/generation) was negative, possibly due to the strong emphasis on selection for BRY and the negative correlations between these two traits. PMID- 19283678 TI - Effect of leptin gene polymorphisms on growth, slaughter and meat quality traits of grazing Brangus steers. AB - Leptin is a hormone that affects the regulation of feed intake, energy balance and body composition in mammals. Several polymorphisms in the bovine leptin gene have been associated with phenotypic variance of these traits. We evaluated two known single nucleotide polymorphisms (SNPs) in the leptin gene of 253 grazing Brangus steers. Brangus is a 5/8 Angus-3/8 Brahman composite. Data were collected during two consecutive growth/fattening cycles from two farms in southeast Buenos Aires province, Argentina. One of the markers is in the promoter region of the gene (SNP1) and the other is a non-synonymous polymorphism in exon 2 (SNP2). The traits that we evaluated were live weight gain in the spring, gain in backfat thickness in the spring, final live weight, final ultrasound backfat thickness, final ultrasound rib eye area, carcass weight and length, carcass yield, kidney fat, kidney fat percentage, backfat thickness, rib eye area, and intramuscular fat percentage. Both markers affected some meat traits; though the only significant associations were of SNP1 with ultrasound rib eye area and of SNP2 with carcass yield and backfat thickness. Under the same conditions as in the present study, leptin markers could be of help only as part of a larger genotyping panel including other relevant genes. PMID- 19283679 TI - Highly conserved regions in the 5' region of human olfactory receptor genes. AB - Regulation of human olfactory receptor (hOR) genes is a complex process of control and signalization with various structures and functions that are not clearly understood. To date, nearly 390 functional hOR genes and 462 pseudogenes have been discovered in the human genome. Enhancer models and trans-acting elements for the regulation of different hOR genes are among the few examples of our knowledge concerning regulation of these genes. We looked for upstream control elements that might help explain these complex control mechanisms. To analyze the human olfactory gene family, we looked for functional genes and pseudogenes common to all hOR genes obtained from public databases. Subsequently, we analyzed sequences upstream of the transcription start sites with data mining and bioinformatics tools. We found two highly conserved regions, which we called HCR I and HCR II, upstream of the transcription start sites in 77 hOR genes and 87 pseudogenes. These regions showed possible enhancer functions common to both genes and pseudogenes, an intriguing feature that may be associated with the expression of pseudogenes. Based on these HCRs, we propose a structural model of gene regulation for the olfactory gene family. PMID- 19283680 TI - Inflammatory response to isocyanates and onset of genomic instability in cultured human lung fibroblasts. AB - Lungs comprise the primary organ exposed to environmental toxic chemicals, resulting in diverse respiratory ailments and other disorders, including carcinogenesis. Carcinogenesis is a multi-stage phenomenon, which involves a series of genetic alterations that begin with genomic instability provoked by certain factors such as inflammation and DNA damage and end with the development of cancer. Isocyanates such as methyl isocyanate are the chief metabolic intermediates in many industrial settings with diverse applications; exposure to them can lead to severe hypersensitive, mutagenic and genotoxic alterations. We examined the molecular mechanisms underlying isocyanate-mediated inflammatory responses and their probable role in the onset of genomic instability in cultured IMR-90 human lung fibroblasts. The isocyanates induced inflammation, resulting in extensive DNA damage, evidenced by increases in ATM, ATR, gammaH2AX, and p53 expression levels. The apoptotic index also increased. Chromosomal anomalies in treated cells included over-expression of centrosome protein and variable amplification of inter-simple sequence repeats, further demonstrating isocyanate induced genomic instability. This information could be useful in the design of new approaches for risk assessment of potential industrial disasters. PMID- 19283681 TI - Molecular markers in commercial Bombyx mori (Lepidoptera: Bombycidae) hybrids susceptible to multiple nucleopolyhedrovirus. AB - The silkworm Bombyx mori L. is particularly susceptible to virus diseases, especially B. mori nucleopolyhedrovirus (BmNPV). Disease resistance, along with high productivity, are important selection criteria for developing commercial hybrids of B. mori. We used bioassays and molecular markers linked to susceptibility/resistance to baculovirus infection to analyze the response of commercial B. mori hybrids from two companies to a geographic isolate of B. mori multiple nucleopolyhedrovirus (BmMNPV) from Parana state in Brazil. Both of these commercial lines were highly susceptible to BmMNPV, with death rates of 92 and 94%. A polymorphic fragment of approximately ~350 bp, associated with susceptibility, and an ~800-bp fragment, associated with resistance to BmMNPV, were detected in all specimens. An additional fragment of ~480 bp, recently identified by our research team as a BmMNPV genomic sequence, was detected in the infected silkworms and could be used as a molecular marker for the diagnosis of nucleopolyhedrovirus infection. PMID- 19283682 TI - Incidence of neural tube defects in Afyonkarahisar, Western Turkey. AB - The incidence of neural tube defects is higher in Turkey compared to that of developed countries. To prevent congenital malformations, understanding of the current status is necessary, which should be followed by public-based activities. We examined the incidence rate of neural tube defects (NTDs) in Afyonkarahisar. According to the records of the Department of Pediatrics, Zubeyde Hanim Hospital for Children's and Women's Health in Afyonkarahisar, the total number of births was 8631 during 2003 and 2004. Sixty-three babies with anomalies were identified in the early postnatal period. The incidence of neural tube defect based on records of hospitals in the city center was calculated as 3.58/1000, among which 9 (1.04%) of the malformed babies had spina bifida, 2 (0.23%) had encephalocele, 12 (1.39%) had anencephaly, and 8 (0.92%) had meningocele/meningomyelocele. In 32 of the 63 cases, there were also other malformations (cleft lip or clubfoot, hydrocephalus, foot abnormalities, etc.). We calculated the total incidence of NTDs, including live births, stillbirths and therapeutic abortions. Stillbirths referred to all fetal deaths after 24 weeks or longer gestation. In each case, the type of anomaly was determined. Thirty-one babies with an NTD were recorded among 8631 gestations (all live births, stillbirths and therapeutic abortions). The incidence of NTDs was found to be 35.9 per 10,000 live births in Afyonkarahisar. The incidence of spina bifida/anencephaly was 0.748 per 1000 newborns. Maternal illiteracy, maternal advanced age and residence in northern or eastern regions of Turkey were found to be risk factors for having a baby with an NTD. The incidence of NTDs is higher than in other European countries. PMID- 19283683 TI - Cytopathology of Bombyx mori (Lepidoptera: Bombycidae) silk gland caused by multiple nucleopolyhedrovirus. AB - A multiple nucleopolyhedrovirus previously isolated from infected Bombyx mori L. larvae (BmMNPV) in Parana state, Brazil, was inoculated into B. mori larvae to examine susceptibility and cytopathology in silk gland cells. The anterior, middle and posterior silk glands were removed from the infected silkworm at different times post-inoculation and processed for cytopathology studies by light and transmission electron microscopy. BmMNPV infection was only detected at 72 h post-inoculation in cells of the middle and posterior silk glands. No sign of infection was found in the anterior silk gland. Cytopathological characteristics were similar to those found in other target tissues; initially, they consisted of nuclear hypertrophy and the formation of virogenic stroma (viroplasm), in which the progeny virions are produced. The virions are then enveloped and occluded in protein crystal structures, the polyhedra. After viral replication, cells undergo lysis and release a great number of BmMNPV polyhedra into the hemocoele. Histopathology showed early infection foci in regions surrounding trachea insertions, demonstrating that trachea is an infection-spreading organ in the insect body. Trachea penetrates the middle and posterior silk gland basal lamina, considered a barrier to virus, facilitating the penetration of budded virus. The anterior silk gland does not have tracheal insertions into the basal lamina, which reduces budded virus infection. Tracheal branches provide a conduit for non occluded virions (budded virus) to pass through the basal lamina barrier and disseminate BmMNPV in the silkworm silk gland. PMID- 19283684 TI - A novel COL1A1 gene-splicing mutation (c.1875+1G>C) in a Brazilian patient with osteogenesis imperfecta. AB - Osteogenesis imperfecta is a heterogeneous genetic disorder characterized by bone fragility and deformity, recurrent fractures, blue sclera, short stature, and dentinogenesis imperfecta. Most cases are caused by mutations in COL1A1 and COL1A2 genes. We present a novel splicing mutation in the COL1A1 gene (c.1875+1G>C) in a 16-year-old Brazilian boy diagnosed as a type III osteogenesis imperfecta patient. This splicing mutation and its association with clinical phenotypes will be submitted to the reference database of COL1A1 mutations, which has no other description of this mutation. PMID- 19283685 TI - A genetic analysis of post-weaning feedlot performance and profitability in Bonsmara cattle. AB - The aim of this study was to identify factors influencing profitability in a feedlot environment and to estimate genetic parameters for and between a feedlot profit function and productive traits measured in growth tests. The heritability estimate of 0.36 for feedlot profitability shows that this trait is genetically inherited and that it can be selected for. The genetic correlations between feedlot profitability and production and efficiency varied from negligible to high. The genetic correlation estimate of -0.92 between feed conversion ratio and feedlot profitability is largely due to the part-whole relationship between these two traits. Consequently, a multiple regression equation was developed to estimate a feed intake value for all performance-tested Bonsmara bulls, which were group fed and whose feed intakes were unknown. These predicted feed intake values enabled the calculation of a post-weaning growth or feedlot profitability value for all tested bulls, even where individual feed intakes were unknown. Subsequently, a feedlot profitability value for each bull was calculated in a favorable economic environment, an average economic environment and in an unfavorable economic environment. The high Pearson and Spearman correlations between the estimate breeding values based on the average economic environment and the other two environments suggested that the average economic environment could be used to calculate estimate breeding values for feedlot profitability. It is therefore not necessary to change the carcass, weaned calf or feed price on a regular basis to allow for possible re-rankings based on estimate breeding values. PMID- 19283686 TI - Allelic variability in the third intron of the fibroin light chain gene in Bombyx mori (Lepidoptera: Bombycidae). AB - Conformation-sensitive gel electrophoresis is a useful method for identifying allele polymorphism; it provides co-dominant molecular markers. Using this method, we identified genetic variability in the third intron of the fibroin light chain gene, fib-L, in six Bombyx mori strains. Only Chinese C21A strain did not demonstrate allelic alterations, showing only homoduplex DNA molecules. We found distinct heteroduplex profiles in the Japanese HAA, M12B and M19-2 and the Chinese C25B and C24-2 strains. Analysis with restriction endonuclease fingerprinting conformation-sensitive gel electrophoresis demonstrated the potential of this method for the identification of allelic variability in B. mori; this was confirmed by cloning and sequencing the different alleles. The main alteration was a 12-bp deletion in two alleles of the C24-2 strain and one allele of the HAA strain; this deletion results in specific heteroduplex DNA molecule profiles. PMID- 19283687 TI - Gender identification of five genera of stingless bees (Apidae, Meliponini) based on wing morphology. AB - Currently, the identification of pollinators is a critical necessity of conservation programs. After it was found that features extracted from patterns of wing venation are sufficient to discriminate among insect species, various studies have focused on this structure. We examined wing venation patterns of males and workers of five stingless bee species in order to determine if there are differences between sexes and if these differences are greater within than between species. Geometric morphometric analyses were made of the forewings of males and workers of Nannotrigona testaceicornis, Melipona quadrifasciata, Frieseomelitta varia, and Scaptotrigona aff. depilis and Plebeia remota. The patterns of males and workers from the same species were more similar than the patterns of individuals of the same sex from different species, and the patterns of both males and workers, when analyzed alone, were sufficiently different to distinguish among these five species. This demonstrates that we can use this kind of analysis for the identification of stingless bee species and that the sex of the individual does not impede identification. Computer-assisted morphometric analysis of bee wing images can be a useful tool for biodiversity studies and conservation programs. PMID- 19283688 TI - Detection of polymorphisms of the mtDNA control region of Caretta caretta (Testudines: Cheloniidae) by PCR-SSCP. AB - Marine turtles are increasingly being threatened worldwide by anthropogenic activities. Better understanding of their life cycle, behavior and population structure is imperative for the design of adequate conservation strategies. The mtDNA control region is a fast-evolving matrilineal marker that has been employed in the study of marine turtle populations. We developed and tested a simple molecular tracing system for Caretta caretta mtDNA haplotypes by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Using this technique, we were able to distinguish the SSCP patterns of 18 individuals of the haplotypes CC-A4, CC-A24 and CCxLO, which are commonly found in turtles sampled on the Brazilian coast. When we analyzed 15 turtles with previously unknown sequences, we detected two other haplotypes, in addition to the other four. Based on DNA sequencing, they were identified as the CC-A17 and CC-A1 haplotypes. Further analyses were made with the sea turtles, Chelonia mydas (N = 8), Lepidochelys olivacea (N = 3) and Eretmochelys imbricata (N = 1), demonstrating that the PCR-SSCP technique is able to distinguish intra- and interspecific variation in the family Cheloniidae. We found that this technique can be useful for identifying sea turtle mtDNA haplotypes, reducing the need for sequencing. PMID- 19283689 TI - Differential mRNA expression of acetylcholinesterase in the central nervous system of rats with acute and chronic exposure of sarin & physostigmine. AB - A time-course study was carried out to measure the acetylcholinesterase (AChE) gene expression in the brain of female rats exposed to different doses of sarin and physostigmine. Short-term effects were studied with an acute single subcutaneous dose (s.c.) of 80 microg kg(-1) (0.5 x LD(50)) sarin. Cortex and cerebellum showed a significant decline in AChE mRNA expression at 2.5, 24 and 72 h. Biochemical studies showed that plasma butrylcholinesterase (BChE) and brain AChE activities were significantly decreased at 2.5 h, which came back to near control values by 24 h in both cases. For long-term chronic studies, three groups of female rats received daily doses of physostigmine (0.1 mg kg(-1) day(-1)) intramuscularly (i.m.), sarin (15 microg kg(-1) day(-1)) s.c. independently and a combined dose of physostigmine (i.m.) (0.1 mg kg(-1) day(-1)) followed by sarin (s.c.) (15 microg kg(-1) day(-1)) continuously for 30 days. Differential AChE mRNA levels in cortex and cerebellum of rat brain were observed after 30 days and after a lag period of another 30 days with no further administration. Plasma (BChE) and brain (AChE) showed irregular inhibition profile in biochemical studies at 30 days and returned to control levels after 60 days. The acute single subcutaneous administration of sarin for short-term as well as chronic long-term studies showed that AChE inhibition alone does not lead to observed changes in mRNA expression of AChE gene. These observations further suggest that route of administration as well as dose exposure regimen also contributes to the regulation of AChE mRNA expression. PMID- 19283690 TI - Parathyroid fine-needle aspiration cytology in the evaluation of parathyroid adenoma: cytologic findings from 53 patients. AB - This study was designed to assess the utility of fine-needle aspiration cytology (FNAC) in the preoperative localization of parathyroid adenoma (PA). Fifty-seven samples from fifty-three cases of PA (four patients had bilateral disease) were obtained by ultrasound (US)-guided fine-needle aspiration. Parathyroid hormone (PTH) estimation was performed on the supernatant of the aspirated fluid on all cases. Subsequently, all of them underwent cytologic evaluation. The cytology slides were evaluated using the following criteria: Cellularity, architectural patterns, bare nuclei in the background, nuclear morphology, and background features (colloid-like material or macrophages). Parathyroid cells were seen in 23 samples (40.4%). The cellularity of the smears was insufficient for interpretation in 16 samples (28.1%); and thyroid follicles and colloid were seen in 18 samples (31.5%). Majority of the samples with parathyroid cells showed moderate cellularity with monomorphous round to slightly oval cells predominantly arranged in loose two-dimensional clusters with occasional papillary fragments. Majority of them exhibited a stippled nuclear chromatin. No significant pleomorphism, mitotic activity, or prominent nucleoli were observed. Most samples showed bare nuclei in the background. In conclusion, US-guided FNAC has its limitations because of low sensitivity in primary localization of the parathyroid adenoma in cases of primary hyperparathyroidism and is not a useful mode of investigation in cases of PA. PMID- 19283691 TI - Optical second harmonic generation chiral spectroscopy. AB - Chiral spectroscopic study: The intensities of second harmonic generation chiral spectroscopy are obtained from the dispersion of the nonlinear optical susceptibility components, as a function of wavelength for helicenbisquinone thin films (see figure). A single formalism fits all the data simultaneously, and the findings constitute an important milestone towards the development of a new experimental technique. PMID- 19283692 TI - 2-Phenylquinolones as inhibitors of the HIV-1 Tat-TAR interaction. AB - Novel 2-phenylquinolones aimed at the Tat-TAR complex were synthesized and tested. Derivatives characterized by precise modifications of the quinolone nucleus and to the side chain of the 2-phenyl ring allowed a thorough structure activity study, confirming 2-phenylquinolone as a suitable scaffold for inhibition of the Tat-TAR interaction. PMID- 19283693 TI - Interactions of HIV-1 antibodies 2F5 and 4E10 with a gp41 epitope prebound to host and viral membrane model systems. AB - Two HIV-1 recognition domains for the human monoclonal antibodies (MAb) 2F5, which recognises the core sequence ELDKWA, and 4E10, which recognises the core sequence NWFNIT, serve as promising models for immunogens in vaccine development against HIV-1. However, the failure of these recognition domains to generate broadly reactive neutralizing antibodies, and the putative membrane-binding properties of the antibodies raised to these recognition domains, suggest that additional features or recognition motifs are required to form an efficient immunogen, which could possibly include the membrane components. In this study we used an extended peptide epitope sequence derived from the gp41 native sequence (H-NEQELLELDKWASLWNWFNITNWLWYIK-NH), which contains the two recognition domains for 2F5 and 4E10, to examine the role of model cell (POPC) and viral (POPC/cholesterol/sphingomyelin) membranes in the recognition of these two antibodies. By using a surface plasmon resonance biosensor, the binding of 2F5 and 4E10 to membranes was compared and contrasted in the presence and absence of prebound peptide epitope. The recognition of the peptide epitope by each MAb was found to be distinct; 2F5 exhibited strong and almost irreversible binding to both membranes in the presence of the peptide, but bound weakly in the absence of the peptide epitope. In contrast, 4E10 exhibited strong membrane binding in the presence or absence of the peptide epitope, and the binding was essentially irreversible in the presence of the peptide epitope. Overall, these results demonstrate that both 2F5 and 4E10 can bind to membranes prior to epitope recognition, but that high-affinity recognition of gp41-derived epitope sequences by 2F5 and 4E10 occurs in a membrane context. Moreover, 4E10 might utilise the membrane to access and bind to gp41; such membrane properties of 2F5 and 4E10 could be exploited in immunogen design. PMID- 19283694 TI - Infrared absorption spectra of t-HNOH radicals generated on VUV irradiation of NO in solid hydrogen. AB - Photoproduct signature: Irradiation of solid hydrogen near 3 K containing NO with vacuum-UV light from synchrotron radiation yields new infrared absorption lines at 1241.7, 1063.6 and 726.2 cm(-1) (see figure). These new lines are assigned to vibrational modes of t-HNOH. This photoproduct is formed from electronically excited NO reacting with neighboring hydrogen in the solid sample.Irradiation of solid H(2) near 3 K containing NO with vacuum-ultraviolet light from a synchrotron yields new infrared absorption lines at 1241.7, 1063.6 and 726.2 cm( 1). The structures of four possible structural isomers: H(2)NO, t-HNOH, c-HNOH and NOH(2), their vibrational wavenumbers, IR intensities and D-isotopic shifts are calculated with density-functional theory according to B3LYP and PW91PW91/aug cc-pVTZ methods. Based on the results of those calculations and of experiments with deuterium labeling, we assign the new lines to nu(4) (cis bending), nu(5) (N==O stretching) and nu(6) (out-of-plane deformation) modes, respectively, of t HNOH. This photoproduct is formed through reaction of electronically excited NO with neighboring H(2) in the solid sample. PMID- 19283695 TI - Microchip frontal affinity chromatography to study the binding of a ligand to teicoplanin-derivatized microbeads. AB - Microchip frontal affinity chromatography was demonstrated to estimate the binding of 5-carboxyfluorescin-(D-Ala)(3) (1) to magnetic microbeads derivatized with teicoplanin (Teic) from Actinoplanes teicomyceticus. In this technique, a cross-chip was used whereby the two side channels contained an identical length (1.5 mm) of derivatized Teic microbeads (affinity column) and underivatized beads (control column), respectively. Cylindrical NdFeB magnets were fabricated into the PDMS chips to retain the magnetic beads. Upon application of a voltage, a sample of 1 was continuously introduced into the affinity column followed by a buffer wash and the same sample from the control channel. The extent of interaction between 1 and the two types of beads in either microchannel resulted in differences in migration time of the ligand as detected by fluorescence. This difference was used to obtain a value for the binding constant between 1 and Teic beads of 5.4 x 10(4) M(-1). This technique reduces the amount of sample needed for the binding assay as compared with conventional frontal affinity chromatography techniques. PMID- 19283696 TI - The autocorrelation matrix probing biochemical relationships after metabolic fingerprinting with CE. AB - Fingerprinting together with statistical analysis is often employed to compare samples in metabonomic studies of a disease. Correlation algorithms can aid by extracting information based on the variation patterns of key metabolites. This information can be linked to metabolite identification or to specific up/down regulated biochemical pathways. Matlab-based software employing the Pearson's correlation algorithm was applied to urine electropherograms from 20 mice infected with the schistosoma parasite. The fingerprints were the sum of electropherograms analysed with normal and reverse polarity, in two different modes MEKC and CZE and with two different capillaries (uncoated and polyacrylamide coated) to provide a broad picture of the samples. Hippurate, a metabolite that was depleted in the infected group and is present in both polarities, was chosen as a test variable; it correlated with itself to a p value of <0.000. Phenylacetylglycine, a metabolite shown as over expressed in the disease, was positively correlated to three metabolites in its same pathway with a correlation coefficient of 0.7 and p<0.000 to phenylalanine, 0.7 and p<0.000 to 2-hydroxyphenylacetic and 0.55 and p<0.003 to phenylacetate. The study shows that the autocorrelation matrix is able to provide extra information from data files acquired by CE analyses. It underlined an up-regulated metabolic path by association in the schistosoma infection model. PMID- 19283697 TI - Untargeted metabolite analysis of healthy and Huanglongbing-infected orange leaves by CE-DAD. AB - Huanglongbing (HLB) is considered the most destructive bacterial citrus disease worldwide. Early detection of HLB is crucial for minimizing its spread. CE was used for the discovery of potential biomarkers for HLB. Optimization of extraction and separation allowed resolving 24 compounds of which 6 were present in significantly higher (p<0.05) concentrations in HLB-infected samples collected monthly for 6 months during the 2007-2008 season. Three of these compounds were identified by mobility and UV spectra as hesperidin, naringenin, and quercetin with mean increase in concentration of 154, 555, and 467%, respectively, above that in healthy leaves. Results support the potential of CE-DAD for untargeted plant metabolomic analysis. CZE, NACE, and MEEKC were compared for metabolic differentiation of healthy and HLB-infected citrus leaves. CZE in a semi-aqueous BGE solution consisting of 8.5 mM of sodium borate (pH 9.3), 15% ACN, and 9% 1 butanol yielded the best peak separation with detection at 190 nm. PMID- 19283698 TI - Enantioselective in-line and off-line CE methods for the kinetic study on cimetidine and its chiral metabolites with reference to flavin-containing monooxygenase genetic isoforms. AB - An in-line screening and an off-line chiral CE method were developed to determine the stereoselectivity of flavin-containing monooxygenase (FMO) isoforms using cimetidine (CIM) as a substrate. The S-oxygenation of CIM was investigated using achiral chemical oxidants and (human supersomes) enzymatic metabolism procedures. In the off-line setup, the chiral selector sulfobutylether-beta-CD was chosen to separate the CIM S-oxide (CSO) metabolites. The electrophoretic migration order of CSO was confirmed to be (+) before (-) through the use of single enantiomers obtained by preparative chromatography. For the electrophoretically mediated microanalysis method, the in-line enzymatic reaction was performed in 100 mM phosphate reaction buffer (pH 8.3), whereas 50 mM phosphate buffer with 30 mM chiral selector (pH 2.5) was used as a BGE. During the screening of FMO isoenzymes by the electrophoretically mediated microanalysis method, formation of the new chiral center on the CIM sulfur was found to be stereoselective. FMO1 produces more (-)-CSO-enantiomer, while FMO3 generates mainly (+)-CSO-enantiomer. On the other hand, FMO5 shows no activity. The kinetic constants of FMO1 and FMO3 were measured by the off-line method. A K(m)=4.31 mM for the formation of the (+) CSO-enantiomer and a K(m)=4.56 mM for the (-)-CSO-enantiomer are reported for the first time for FMO1. PMID- 19283699 TI - Comparison of 1-D and 2-D LC MS/MS methods for proteomic analysis of human serum. AB - 1-D and 2-D LC methods were utilized for proteome analysis of undepleted human serum. Separation of peptides in 2-D LC was performed either with strong cation exchange (SCX)-RP chromatography or with an RP-RP 2-D LC approach. Peptides were identified by MS/MS using a data-independent acquisition approach. A peptide retention prediction model was used to highlight the potential false-positive peptide identifications. When applying selected data filtration, we identified 52 proteins based on 316 peptides in serum in 1-D LC setup. One hundred and eighty four proteins/1036 peptides and 142 proteins/905 peptides were identified in RP RP and SCX-RP 2-D LC, respectively. The performance of both 2-D LC methods for proteomic analysis is critically compared. PMID- 19283704 TI - Mouse Th17 cells: current understanding of their generation and regulation. AB - IL-17-expressing CD4(+) T cells have been recently recognized as a new subset of Th cells, namely Th17 cells. Considerable progress has been made in understanding the developmental regulation of mouse Th17 cells. Here, I summarize this knowledge and discuss on the relationship of Th17 with regulatory and follicular Th cells. PMID- 19283705 TI - IL-17/Th17 in anti-fungal immunity: what's new? AB - How the immune system tailors protective responses to suit the infectious challenge while limiting damage to the host is an emerging theme in T-cell biology. Although many studies have focused on the pathological aspects of IL-17 producing T cells in many autoimmune diseases, their role in protective anti microbial immunity has also been increasingly recognized. This increased recognition also applies to their role in anti-fungal immunity; however, the role of IL-17-producing T cells in protection versus pathology in fungal infections is still controversial. Although both positive and negative effects on immune resistance have been attributed to the IL-23/Th17 axis in experimental models of fungal infections, defective Th17 cell differentiation has been linked to recurrent pneumonia by filamentous fungi and the occurrence of mucocutaneous candidiasis in patients with primary immunodeficiencies. Here we discuss how recent findings in experimental candidiasis and aspergillosis shed new lights on the contribution of Th17 cells to resistance and pathology to fungi. PMID- 19283706 TI - IL-17 and anti-bacterial immunity: protection versus tissue damage. AB - IL-17 can impact health in a variety of ways. It is protective for some pathogens but it is also associated with tissue damaging inflammation. By examining the role of IL-17 in a variety of bacterial infections the mechanisms by which this cytokine mediates both protection and damage can be dissected. A key element in understanding the role of this cytokine is determining where and when it is acting. Dissecting its essential protective role from its immunopathologic role will allow for improved intervention in both acute and chronic disease. PMID- 19283707 TI - BM stromal cells ameliorate experimental autoimmune myasthenia gravis by altering the balance of Th cells through the secretion of IDO. AB - In addition to their capacity to differentiate, BM stromal cells (BMSC) have immunosuppressive qualities that make them strong candidates for use in cell therapy against human autoimmune diseases. We studied the immunoregulatory activities of BMSC on experimental autoimmune myasthenia gravis (EAMG) in vitro and in vivo. Intravenous administration of syngenic BMSC to EAMG-model rats on the day of their second immunization was effective in ameliorating the pathological features of the disease. In vitro, the proliferative ability of T cells or B cells from EAMG rats was inhibited when they were cocultured with BMSC at proper ratios. This inhibitory effect was at least partially dependent on the secretion of IDO. We also determined that the development of EAMG is accompanied by an imbalance among the Th1, Th2, Th17, and Treg cell subsets, and that this can be corrected by the administration of BMSC, which leads to an increase of Th2 (IL-4) and Treg (Foxp3) cells, and a reduction of Th1 (IFN-gamma) and Th17 (IL 17) cells, through an IDO-dependent mechanism. These results provide further insights into the pathogenesis of MG, EAMG, and other immune-mediated diseases, and support a potential role for BMSC in their treatment. PMID- 19283709 TI - p21(waf1/CIP1), a CDK inhibitor and a negative feedback system that controls macrophage activation. AB - p21(WAF1/CIP1) (p21) is a crucial CDK inhibitor that controls the cell cycle. This molecule is also involved in the regulation of apoptosis and gene expression. However, like many other cell regulators, the functional activity of p21 depends on its cellular context and is controlled through phosphorylation and protein-protein interactions. p21 is also important in cells of the immune system regulating the cell cycle and preventing apoptosis of macrophages. In this issue of the European Journal of Immunology, two reports investigate the role of p21 further determining its critical role as a negative regulator of macrophage activation, in particular inhibiting the LPS-dependent induction of TNF-alpha and IL-1beta. The inhibition mediated by p21 is shown to be related to NF-kappaB activity. Furthermore, the observation that p21(-/-) mice are more susceptible to septic shock supports the notion that p21 is a negative regulator of macrophage activation and therefore a potential new target to control inflammatory diseases. PMID- 19283711 TI - The day of immunology 2009. PMID- 19283712 TI - Host versus pathogen: two sides of the same challenge in the TB world. PMID- 19283713 TI - Th17: The ascent of a new effector T-cell subset. Preface. PMID- 19283714 TI - Human Th17 cells: are they different from murine Th17 cells? AB - Type 17 Th (Th17) cells have been identified as a distinct population of CD4(+) effector T cells different from Th1 and Th2 cells. While the pre-eminent cytokine of Th1 cells is IFN-gamma and that of Th2 cells is IL-4, the distinctive cytokine of Th17 cells is IL-17A. However, although murine and human Th1 and Th2 cells exhibit strong similarities, human and murine Th17 cells seem to differ in several aspects. PMID- 19283715 TI - Modulation of Th17 development and function by activation of the aryl hydrocarbon receptor--the role of endogenous ligands. AB - The elucidation of the crucial differentiation factors for the new Th17 CD4 effector T-cell subset spurred an explosive growth in research publications focused on these cells and led to rapid advances in knowledge concerning their regulation and functional activities. Here we discuss recent discoveries linking the development and functional potential of Th17 cells to a transcription factor that mediates the response to exogenous and endogenous environmental signals. PMID- 19283716 TI - Environmental control of Th17 differentiation. AB - Th17 cells participate in the control of extracellular bacteria and fungi, but dysregulated Th17 cell activity can result in immunopathology. Thus, the generation and the activity of Th17 cells should be tightly regulated, and simultaneously it should be able to respond to the challenges presented by a changing environment. In this viewpoint, we discuss some of the mechanisms by which microbes, dietary components and environmental toxins influence the Th17 response. PMID- 19283717 TI - Cytokine mediators of Th17 function. AB - Th17 cells were identified as an independent lineage of CD4(+) T cells that secrete a distinctive set of immunoregulatory cytokines, including IL-17A, IL 17F, IL-22, and IL-21. These cytokines collectively play roles in inflammation and autoimmunity and in response to extracellular pathogens. The expression of the lineage-specific transcription factor RORgammat leads to Th17 lineage commitment; however, it has become increasingly clear that the population of cells designated as Th17 cells is not homogeneous. Although these cells collectively produce characteristic Th17 cytokines, not all are produced by each individual cell in the population. The cytokines produced by individual cells are presumably affected in part by the specific local cytokine milieu. In this review, we discuss the current understanding of the specific functional characteristics and regulation of Th17 cytokines and clarify how they mediate the actions of Th17 cells. PMID- 19283718 TI - IL-17-producing gammadelta T cells. AB - IL-17 is produced not only by CD4(+) alphabeta T cells, but also CD8(+) alphabeta T cells, NKT cells, and gammadelta T cells, plus some non-T cells, including macrophages and neutrophils. The ability of IL-17 to deploy neutrophils to sites of inflammation imparts this cytokine with a key role in diseases of several types. Surprisingly, gammadelta T cells are responsible for much of the IL-17 produced in several disease models, particularly early on. PMID- 19283719 TI - Diseases that may benefit from manipulating the Th17 pathway. AB - The discovery of IL-17 and of the Th17 pathway has been a step in the classification of human diseases. Th17 targeting appears rather straightforward in diseases associated with inflammation and matrix destruction, such as rheumatoid arthritis, psoriasis, and Crohn's disease. In other conditions where IL-17 is expressed and Th17 activated, it is unclear whether this is a primary or secondary event, making the use of Th17 inhibitors less obvious. PMID- 19283720 TI - Novel therapeutic targets along the Th17 pathway. AB - The recent discovery of IL-17-producing CD4(+) Th subset significantly revised the Th1/Th2 dichotomy model proposed by Mosmann and Coffman almost two decades ago. Th17 cells are involved in the pathogenesis of many human autoimmune diseases. Th17 cells, their developmental pathways and their effector functions, therefore, provide novel therapeutic targets. PMID- 19283721 TI - How B cells shape the immune response against Mycobacterium tuberculosis. AB - Extensive work illustrating the importance of cellular immune mechanisms for protection against Mycobacterium tuberculosis has largely relegated B-cell biology to an afterthought within the tuberculosis (TB) field. However, recent studies have illustrated that B lymphocytes, through a variety of interactions with the cellular immune response, play previously underappreciated roles in shaping host defense against non-viral intracellular pathogens, including M. tuberculosis. Work in our laboratory has recently shown that, by considering these lymphocytes more broadly within their variety of interactions with cellular immunity, B cells have a significant impact on the outcome of airborne challenge with M. tuberculosis as well as the resultant inflammatory response. In this review, we advocate for a revised view of TB immunology in which roles of cellular and humoral immunity are not mutually exclusive. In the context of our current understanding of host defense against non-viral intracellular infections, we review recent data supporting a more significant role of B cells during M. tuberculosis infection than previously thought. PMID- 19283722 TI - CTLA-4 co-receptor impacts on the function of Treg and CD8+ T-cell subsets. AB - CTLA-4 has potent regulatory effects on the threshold of T-cell signalling and, in the process, guards against the development of hyper-proliferation and autoimmunity. Despite this, the role of CTLA-4 on specific T-cell subsets has been unclear. Such studies could shed light on both the function of CTLA-4, and on the contribution of the subsets to the disease phenotype of the Ctla4(-/-) mouse. Recently, a role for this co-receptor in the function of Treg has been outlined and, in this issue of the European Journal of Immunology, the selective targeting of the T-box transcription factor Eomes by CTLA-4 in the regulation of CD8(+) cytolytic T-cell (CTL) effector function is shown. Together, these papers shed light on the role of CTLA-4 in different T-cell subsets. PMID- 19283723 TI - Hydrogen/deuterium exchange mass spectrometry identifies two highly protected regions in recombinant full-length prion protein amyloid fibrils. AB - Understanding the structural basis that distinguishes the amyloid form of the prion protein from its monomeric homologue is of crucial importance to elucidate the mechanism of the lethal diseases related to this protein. Recently, an in vitro conversion system was established which reproduces the transition of recombinant prion protein PrP(23-230) from its native alpha-helical rich form into an aggregated amyloid beta-sheet rich form with physicochemical properties reminiscent to those of the disease-related isoform of the prion protein, PrPSc. To study the tertiary and quaternary structural organization within recombinant amyloid fibrils from mouse, mPrP(23-231)betaf; bovine, bPrP(23-230)betaf; and elk, ePrP(23-230)betaf; we utilized hydrogen/deuterium (H/D) exchange analyzed by matrix-assisted laser desorption/ionization (MALDI) and nano-electrospray (nano ESI) mass spectrometry. No significant differences were found by measuring the deuterium exchange kinetics of the aggregated fibrillar forms for mPrP(23 231)betaf, bPrP(23-230)betaf and ePrP(23-230)betaf, indicating a similar overall structural organization of the fibrils from all three species. Next, we characterized the solvent accessibility for the soluble and fibrillar forms of the mouse prion protein by hydrogen exchange, pepsin proteolysis and nano-ESI ion trap mass spectrometry analysis. In its amyloid form, two highly protected regions of mPrP(23-231) comprising residues [24-98] and [182-212] were identified. The residues between the two highly protected stretches were found to be more solvent exposed, but less than in the soluble protein, and might therefore rather form part of a fibrillar interface. PMID- 19283724 TI - Pharmacokinetic interaction of single dose of piperine with steady-state carbamazepine in epilepsy patients. AB - Piperine, the active principle of piper species, is commonly used as a spice and adjuvant in various traditional systems of medicine. It has been known as a bioavailability-enhancer. The present study aimed at evaluating the effect of piperine on the steady-state pharmacokinetics of a single dose of carbamazepine in poorly controlled epilepsy patients on carbamazepine monotherapy. Patients (n = 10 each) receiving either 300 mg or 500 mg dose of carbamazepine twice daily were selected. After administration of carbamazepine, venous blood samples were collected at 0, 0.5, 1, 2, 4, 6, 9, 12 h. Subsequently, piperine (20 mg p.o.) was administered along with carbamazepine and samples were collected similarly. The pharmacokinetic parameters were compared by Students t-test. Piperine significantly increased the mean plasma concentrations of carbamazepine at most of the time points in both dose groups. There was a significant increase in AUC(0 12hr) (p < 0.001), average C(ss) (p < 0.001), t(1?2el) (p < 0.05) and a decrease in K(el) (p < 0.05), in both the dose groups, whereas changes in K(a) and t(1?2a) were not significant. Cmax (p < 0.01) and t(max) (p < 0.01) were increased significantly following piperine administration in the 500 mg dose group; however, these parameters were not significant in the lower dose group. Piperine could significantly enhance the oral bioavailability of carbamazepine, possibly by decreasing the elimination and/or by increasing its absorption. PMID- 19283725 TI - Electrostatic binding of nanoparticles to mesenchymal stem cells via high molecular weight polyelectrolyte chains. AB - Combining stem cell transplantation with nanoparticle-mediated delivery of drugs and pharmaceuticals is envisioned to be one of the next major developmental steps in regenerative medicine. However, a major challenge would be to keep nanoparticles co-localized with stem cells upon transplantation or transfusion in situ. Since nanoparticles are physically much smaller in size than cells and would not specifically bind to extracellular matrix, it is easier for them to disperse from the transplantation site via the blood circulation. Conjugating nanoparticles directly to the cell membrane can potentially interfere with cellular function by physically obstructing cell surface receptors from interacting with the extracellular matrix, various growth factors and cytokines and other cells. Moreover, drug-loaded nanoparticles may be internalized into the cytoplasm via endocytosis or phagocytosis, which may wreak damage on the cellular machinery, leading to impaired physiological function or cell death. A novel solution may be to utilize high molecular weight polyelectrolyte chains to electrostatically bind nanoparticles to cells. For this purpose, hyaluronan, poly L-lysine and chitosan are of special interest, because these molecules are generally recognized to be biocompatible for application in various pharmaceutical and surgical products. This study investigated the use of these molecules to bind nanoparticles to mesenchymal stem cells (MSCs), and a novel technique of conjugating half the cell surface with nanoparticles through the use of polyelectrolyte chains was also developed. This would avoid blocking MSC interaction with cytokines, growth factors, extracellular matrix and other cells within the recipient tissue/organ upon delivery in situ. PMID- 19283726 TI - Responsiveness of human bone marrow stromal cells to shear stress. AB - We examined the hypothesis that human mesenchymal stem cells detect physiological mechanical signals. Human bone marrow stromal cells (HBMSCs) were exposed to fluid shear stress of 12 dynes/cm(2) and analysed for their ability to express osteoblast-specific markers and associated signalling pathways. HBMSCs showed a significant increase in alkaline phosphatase (ALP) gene expression and a marked decrease in type I collagen, while no effect on Cbfa1/Runx2 was detected. This regulation is related to p38 and ERK1/2 activation, although the use of specific inhibitors to these two MAP kinases suggests that ALP mRNA induction is especially dependent on p38 activity, while type I collagen downregulation is ERK1/2-dependent. Interestingly, the expression of connexin43, which is involved in cell-to-cell communication of osteoblastic cells through gap junction formation, and its distribution through the cells, were modified by fluid flow (FF). HBMSCs are sensitive to shear stress and it appears essential to take their responsiveness into consideration before associating these regenerative cells with a bioactive biomaterial in a new bone tissue-engineering strategy. PMID- 19283727 TI - In vivo vascular tissue engineering: influence of cytokine and implant location on tissue specific cellular recruitment. AB - In vivo tissue engineering has been explored as a means to create autologous vascular replacements. Elastin is necessary to sustain continual pulsatile flow and to prevent the dilatation of vascular tissues. Unfortunately, elastogenesis in tissue-engineered constructs has been very limited. To overcome this limitation, we have created tubular elastin scaffolds from porcine carotid arteries. Elastin would provide the necessary elasticity to the graft on implanting these scaffolds as vascular grafts. In this study, elastin tubes with agarose gel containing either stromal-derived factor-1 alpha [SDF; for homing of endothelial cells (ECs)] or basic fibroblast growth factor (bFGF; for homing of myofibroblasts) were implanted into adipose tissue, as it is a known source of stem/progenitor cells. We also implanted these tubes into subdermal pouches (as a control location). We observed a difference in the types of cells recruited-ECs were recruited in large numbers by SDF in the adipose tissue, whereas the adipose FGF group had a vascularized (smooth muscle and EC-positive), collagenous capsule (adventitia) with many smooth muscle alpha-actin (SMA)-positive cells in the elastin scaffold layer (media). These results were in contrast to the subdermal group, which only recruited fibroblasts and some SMA-positive cells. Also, more cell infiltration and neo-collagen formation was seen in adipose implants. This study provides novel results by the use of specific cytokines and implant locations to recruit tissue-specific cells to create autologous vascular grafts. PMID- 19283728 TI - Overview: engineering transgenic constructs and mice. AB - Cell biology research encompasses everything from single cells to whole animals. Recent discoveries concerning particular gene functions can be applied to the whole animal for understanding genotype-phenotype relationships underlying disease mechanisms. For this reason, genetically manipulated mouse models are now considered essential to correctly understand disease processes in whole animals. This unit reviews the basic mouse technologies used to generate conventional transgenic mice, which represent a gain-of-function approach. First, an overview of transgenic construct design is presented. This unit then explains basic strategies for the identification and establishment of independent transgenic mouse lines, followed by comments on historical and emerging techniques. It then describes typical problems that are encountered when researchers start to generate transgenic mice. PMID- 19283729 TI - Generation of transgenic mice. AB - This unit describes detailed step-by-step protocols, reagents, and equipment required for successful generation of transgenic mice using pronuclear injection. The experimental methods and practical tips given here will help guide beginners in understanding what is required and what to avoid in these standard protocols for efficiently generating transgenic mice. PMID- 19283731 TI - Fabrication and application of nanofibrous scaffolds in tissue engineering. AB - Nanofibers fabricated by electrospinning are morphological mimics of fibrous components of the native extracellular matrix, making nanofibrous scaffolds ideal for three-dimensional cell culture and tissue engineering applications. Although electrospinning is not a conventional technique in cell biology, the experimental setup may be constructed in a relatively straightforward manner, and the procedure can be carried out by individuals with limited engineering experience. Here, we detail a protocol for electrospinning of nanofibers and provide relevant specific details concerning the optimization of fiber formation (Basic Protocol 1). The protocol also includes conditions required for preparing biodegradable polymer solutions for the fabrication of nonwoven and aligned nanofibrous scaffolds suitable for various cell/tissue applications. In addition, information on effective cell loading into nanofibrous scaffolds and cellular constructs grown in a bioreactor is provided (Basic Protocol 2). Instructions for building the electrospinning apparatus are also included (see the Support Protocol). PMID- 19283730 TI - Visualization of cellular phosphoinositide pools with GFP-fused protein-domains. AB - This unit describes the method of following phosphoinositide dynamics in live cells. Inositol phospholipids have emerged as universal signaling molecules present in virtually every membrane of eukaryotic cells. Phosphoinositides are present in only tiny amounts as compared to structural lipids, but they are metabolically very active as they are produced and degraded by the numerous inositide kinase and phosphatase enzymes. Phosphoinositides control the membrane recruitment and activity of many membrane protein signaling complexes in specific membrane compartments, and they have been implicated in the regulation of a variety of signaling and trafficking pathways. It has been a challenge to develop methods that allow detection of phosphoinositides at the single-cell level. The only available technique in live cell applications is based on the use of the same protein domains selected by evolution to recognize cellular phosphoinositides. Some of these isolated protein modules, when fused to fluorescent proteins, can follow dynamic changes in phosphoinositides. While this technique can provide information on phosphoinositide dynamics in live cells with subcellular localization, and it has rapidly gained popularity, it also has several limitations that must be taken into account when interpreting the data. This unit summarizes the design and practical use of these constructs and also reviews important considerations for interpretation of the data obtained by this technique. PMID- 19283732 TI - BK virus (BKV): infection, propagation, quantitation, purification, labeling, and analysis of cell entry. AB - BK virus (BKV) can cause BKV nephritis in renal transplant patients and has become a significant reason for graft loss in this decade. BKV is latent in the urogenital tract and most likely is transported with the donor kidney to recipients. BKV replication occurs in the nucleus of human renal proximal tubular cells (HRPTEC) and daughter viruses are delivered to other cells to spread infection. A few in vitro studies have been reported about the mechanism and kinetics of BKV infection. However, there are still a lot of unknown factors regarding BKV infection. This unit describes the handling of BKV, BKV propagation, determination of titer and ability to infect cells, as well as purification and labeling of BKV in order to analyze BKV cell entry. PMID- 19283733 TI - Isolation of dense core secretory vesicles from pancreatic endocrine cells by differential and density gradient centrifugation. AB - Methods are presented for the separation of dense core secretory vesicles from insulin-secreting tissues (insulin granules) based on a combination of differential and density gradient centrifugation on various media. Emphasis is given to the use of transplantable tumors, tissue culture cell lines, and pancreatic islets as a tissue source. PMID- 19283735 TI - Outcomes following Turnbull-Cutait abdominoperineal pull-through compared with coloanal anastomosis. AB - BACKGROUND: The Turnbull-Cutait abdominoperineal pull-through procedure (T-C) is used as a last resort to avoid permanent diversion in patients with complex anorectal conditions. The aim was to evaluate short- and long-term outcomes after T-C. METHODS: Patients undergoing T-C from 1996 to 2007 were reviewed retrospectively in terms of demographics, diagnosis, indications and postoperative complications. Patients were contacted to obtain functional outcomes using a standardized questionnaire. Functional outcomes were compared with those in a matched group of patients undergoing handsewn coloanal anastomosis (CAA) for rectal cancer. RESULTS: Sixty-seven patients (40 men) underwent T-C. Postoperative complications included stricture in 11 patients (16 per cent), fistula in five (7 per cent), prolapse of the colon in five (7 per cent) and leak in two (3 per cent). Mean follow-up was 5.6 (s.d. 3.2) years. The operation failed in 17 patients (25 per cent). Among 44 patients (66 per cent) who completed questionnaires, faecal (P = 0.121) and urinary (P = 0.073) incontinence, and sexual function (P = 0.063) were comparable to those in patients who had CAA. CONCLUSION: T-C is an option for patients with complex anorectal conditions that might otherwise require permanent diversion. Functional outcomes are comparable to those of CAA. PMID- 19283736 TI - Randomized clinical trial of Entonox versus midazolam-fentanyl sedation for colonoscopy. AB - BACKGROUND: Intravenous sedation for colonoscopy is associated with cardiorespiratory complications and delayed recovery. The aim of this randomized clinical trial was to compare the efficacy of Entonox (50 per cent nitrous oxide and 50 per cent oxygen) and intravenous sedation using midazolam-fentanyl for colonoscopy. METHODS: Some 131 patients undergoing elective colonoscopy were included. Patients completed a Hospital Anxiety and Depression questionnaire, letter cancellation tests and pain scores on a 100-mm visual analogue scale before, immediately after the procedure and at discharge. They also completed a satisfaction survey at discharge and 24 h after the procedure. RESULTS: Sixty five patients were randomized to receive Entonox and 66 to midazolam-fentanyl. Completion rates were similar (94 versus 92 per cent respectively; P = 0.513). Patients receiving Entonox had a shorter time to discharge. They reported significantly less pain (mean score 16.7 versus 40.1; P < 0.001), and showed better recovery of psychomotor function immediately after the procedure and at discharge. Patient satisfaction was higher among patients who received Entonox (median score 96 versus 89; P = 0.001). CONCLUSION: Entonox provides better pain relief and faster recovery than midazolam-fentanyl and so is more effective for colonoscopy. PMID- 19283737 TI - Endocavitary contact radiation therapy for ultrasonographically staged T1 N0 and T2 N0 rectal cancer. AB - BACKGROUND: The purpose of this study was to determine the long-term outcomes of patients undergoing endocavitary contact radiation therapy (ECR) for stage I rectal cancer. METHODS: A database of patients treated with ECR for biopsy-proven rectal adenocarcinoma from July 1986 to June 2006 was reviewed retrospectively. Only patients with primary, non-metastatic, ultrasonographically staged T1 N0 and T2 N0 cancer who had no adjuvant treatment were included. Patients received a median of 90 (range 60-190) Gy contact radiation, delivered transanally by a 50 kV X-ray tube in two to five fractions. RESULTS: Of 149 patients, 77 (40 T1, 37 T2) met the inclusion criteria. Median age was 74 (range 38-104) years, and median follow-up 69 (range 10-219) months. ECR failed in 21 patients (27 per cent) (persistent disease, four; recurrence, 17), of whom ten remained disease free after salvage therapy. The estimated 5-year disease-free survival rate was 74 (95 per cent confidence interval 63 to 83) per cent after ECR alone, and 87 (76 to 93) per cent when survival after salvage therapy for recurrence was included. CONCLUSION: ECR is a minimally invasive treatment option for early stage rectal cancer. However, similar to other local therapies, ECR has a worse oncological outcome than radical surgery. PMID- 19283738 TI - Mild hepatic macrovesicular steatosis may be a risk factor for hyperbilirubinaemia in living liver donors following right hepatectomy. AB - BACKGROUND: The aim of this study was to evaluate the effects of mild macrovesicular steatosis on the outcome of living liver donors following right hepatectomy. METHODS: The medical records of 46 living liver donors who underwent right hepatectomy were studied. Ten donors had mild macrovesicular steatosis (5 10 per cent in seven and 11-20 per cent in three patients). Five donors with other liver pathology were excluded. Outcome in these ten donors (group 1) was compared with that in the remaining 31 donors with normal liver histology (group 2). RESULTS: The median duration until normalization of total bilirubin levels was 14 and 5 days in groups 1 and 2 respectively (P = 0.028). The peak total bilirubin level was significantly higher in group 1 than in group 2 (80.4 versus 49.6 micromol/l; P = 0.033). Multivariable analysis showed mild macrovesicular steatosis to be an independent risk factor for hyperbilirubinaemia (odds ratio 7.94 (95 per cent confidence interval 1.17 to 54.03); P = 0.034). CONCLUSION: Mild macrovesicular steatosis may be related to adverse outcome in living liver donors who undergo right hepatectomy and, in terms of donor safety, is of potential concern in donor selection. PMID- 19283739 TI - Long-term outcomes of revisional surgery following laparoscopic fundoplication. AB - BACKGROUND: A small proportion of patients who have laparoscopic antireflux procedures require revisional surgery. This study investigated long-term clinical outcomes. METHODS: Patients requiring late revisional surgery following laparoscopic fundoplication for gastro-oesophageal reflux were identified from a prospective database. Long-term outcomes were determined using a questionnaire evaluating symptom scores for heartburn, dysphagia and satisfaction. RESULTS: The database search found 109 patients, including 98 (5.6 per cent) of 1751 patients who had primary surgery in the authors' unit. Indications for surgical revision were dysphagia (52 patients), recurrent reflux (36), mechanical symptoms related to paraoesophageal herniation (16) and atypical symptoms (five). The median time to revision was 26 months. Outcome data were available for 104 patients (median follow-up 66 months) and satisfaction data for 102, 88 of whom were highly satisfied (62.7 per cent) or satisfied (23.5 per cent) with the outcome. Patients who had revision for dysphagia had a higher incidence of poorly controlled heartburn (20 versus 2 per cent; P = 0.004), troublesome dysphagia (16 versus 6 per cent; P = 0.118) and a lower satisfaction score (P = 0.023) than those with recurrent reflux or paraoesophageal herniation. CONCLUSION: Revisional surgery following laparoscopic fundoplication can produce good long-term results, but revision for dysphagia has less satisfactory outcomes. PMID- 19283740 TI - Prognostic value of subclassification of T2 tumours in patients with gastric cancer. AB - BACKGROUND: This study was designed to evaluate the prognostic value of tumour stage T2 subcategorization (T2a and T2b) in patients with gastric carcinoma. METHODS: Clinicopathological details of a prospective series of patients who had radical resection of gastric adenocarcinoma in a single institution were analysed. Univariable and multivariable survival analyses were performed with the log rank test and Cox's model respectively. RESULTS: Of 373 evaluable patients, 49 (13.1 per cent) had a T2a and 143 (38.3 per cent) a T2b tumour. At a median follow-up of 35.5 months, the 5-year overall survival rate was 73 and 31.1 per cent for patients with T2a and T2b lesions respectively (P < 0.001). On multivariable analysis, T stage remained an independent prognostic factor. Compared with T1a, the mortality risk for patients with T1b (hazard ratio (HR) 1.00; P = 0.992) and T2a (HR 0.97; P = 0.916) tumours was similar; by contrast, the risk of death associated with T2b (HR 1.81; P = 0.031) and T3 (HR 1.89; P = 0.038) lesions was significantly greater than for T1a tumours. CONCLUSION: Subclassification of T2 tumours should be undertaken routinely in order to stratify patients with gastric cancer more accurately in terms of their mortality risk. PMID- 19283741 TI - Validation of a nomogram for predicting survival after resection for adenocarcinoma of the pancreas. AB - BACKGROUND: Nomograms are statistical tools providing the overall probability of a specific outcome; they have shown better individual discrimination than the tumour node metastasis staging system in several cancers. The pancreatic nomogram, originally developed in the Memorial Sloan-Kettering Cancer Center (MSKCC) in the USA, combines clinicopathological and operative data to predict disease-specific survival at 1, 2 and 3 years from initial resection. METHODS: An external patient cohort from a retrospective pancreatic adenocarcinoma database at the Academic Medical Centre in Amsterdam was used to test the validity of the pancreatic adenocarcinoma nomogram. The cohort included 263 consecutive patients who had surgery between January 1985 and December 2004. RESULTS: Data for all the necessary variables were available for 256 patients (97.3 per cent). At the last follow-up, 35 patients were alive, with a median follow-up of 27 (range 3-114) months. The 1-, 2- and 3-year disease-specific survival rates were 60.8, 30.4 and 16.0 per cent respectively. The nomogram concordance index was 0.61. The calibration analysis of the model showed that the predicted survival did not significantly deviate from the actual survival. CONCLUSION: The MSKCC pancreatic cancer nomogram provided an accurate survival prediction. It may aid in counselling patients and in stratification of patients for clinical trials. PMID- 19283742 TI - Meta-analysis of standard, restrictive and supplemental fluid administration in colorectal surgery. AB - BACKGROUND: Optimal fluid therapy for colorectal surgery remains uncertain. METHODS: A simple model was applied to define standard, restrictive and supplemental fluid administration. These definitions enabled pooling of data from different trials. Randomized controlled trials on fluid amount (standard versus restrictive or supplemental amount) and on guidance for fluid administration (goal-directed fluid therapy by oesophageal Doppler-derived variables versus conventional haemodynamic variables) in patients with colorectal resection were eligible for inclusion. The primary outcome measure was postoperative morbidity. Secondary endpoints were mortality, cardiopulmonary morbidity, wound infection, anastomotic failure, recovery of bowel function and hospital stay. A random effects model was applied. RESULTS: Nine randomized controlled trials were included. Restrictive fluid amount (odds ratio (OR) 0.41 (95 per cent confidence interval (c.i.) 0.22 to 0.77); P = 0.005) and goal-directed fluid therapy by means of oesophageal Doppler-derived variables (OR 0.43 (95 per cent c.i. 0.26 to 0.71); P = 0.001) significantly reduced overall morbidity. There were no significant differences in the secondary endpoints analysed. CONCLUSION: Using standardized definitions, this meta-analysis suggests that restrictive rather than standard fluid amount according to current textbook opinion, and goal directed fluid therapy rather than fluid therapy guided by conventional haemodynamic variables, reduce morbidity after colorectal resection. PMID- 19283743 TI - Prognosis of synchronous bilateral breast cancer. AB - BACKGROUND: The prognosis of patients with synchronous bilateral breast cancer (SBBC) is usually based on the tumour with the worst pathological features. There is little evidence in the literature for this assumption, potentially impairing reasoned decisions on optimal adjuvant therapy. METHODS: This was a case-control study in which 68 women with SBBC were matched with 128 women with unilateral breast cancer. Both the GuysRisk prognostic model and the Nottingham Prognostic Index were used to determine the bilateral tumour with the poorer prognosis. Controls were matched for age, menopausal status, date of diagnosis, histological type and grade, and oestrogen receptor and axillary node status. RESULTS: Both prognostic models indicated the same side tumour with the worst prognosis. Kaplan Meier survival curves for both disease-free and overall survival showed no significant difference in outcome between the two groups. CONCLUSION: Prognosis was determined by the tumour with the worst prognosis, with no additional worsening of outcome incurred from the second tumour. PMID- 19283744 TI - Cyclical ischaemic preconditioning modulates the adaptive immune response in human limb ischaemia-reperfusion injury. AB - BACKGROUND: Reperfusion injury (RI) has significant local and systemic consequences. Ischaemic preconditioning (IPC) modulates RI and the innate immune response. This study examined whether IPC attenuates RI-mediated changes in lymphocyte populations and function following elective surgery. METHODS: Twenty five patients sustaining 1 h of tourniquet ischaemia during cruciate ligament reconstruction were randomized before surgery to three 5-min ischaemia cycles separated by 5 min of reperfusion, or to a control group. Systemic levels of interleukin (IL) 4 and interferon (IFN) gamma, and surface expression of CD45ro/ra, CD62L and CD95 were measured. T cells were examined systemically and in stimulated serum co-culture to determine CD4/CD8 and Th1/Th2 shifts through intracellular cytokine production. RESULTS: CD4 CD45ro cell numbers increased after RI without IPC, whereas CD8 cells expressing CD45ro and CD95 increased with IPC. Preconditioned serum in co-culture attenuated increases in CD4 and decreases in CD8 numbers, a process prevented by inhibition of antigen activation. Following RI, systemic IL-2 levels were significantly lower after IPC, whereas co culture with post-RI serum increased proinflammatory intracellular cytokine production. CONCLUSION: IPC modulated T cell responses in limb RI through reduced activation and proinflammatory cytokine production by CD4 cells, while preventing CD4/CD8 derangement. IPC prevented lymphocyte-directed immune dysfunction. PMID- 19283745 TI - Randomized clinical trial of concomitant or sequential phlebectomy after endovenous laser therapy for varicose veins. AB - BACKGROUND: The management of residual varicosities following endovenous laser therapy (EVLT) for varicose veins is contentious. Ambulatory phlebectomy may be performed concomitantly with the initial EVLT, or sequentially as a secondary procedure. This randomized trial compared these two approaches. METHODS: Fifty patients with great saphenous varicose veins were randomized to EVLT alone or EVLT with concomitant ambulatory phlebectomies (EVLTAP). Principal outcomes were procedure duration, pain scores, requirement for secondary procedures and quality of life after 3 months. RESULTS: EVLTAP took longer, but significantly decreased the requirement for subsequent interventions. There was no impairment in immediate postprocedural pain, Short Form 36 or EuroQol 5D scores with EVLTAP. Median (i.q.r.) Venous Clinical Severity Score (VCSS) at 3 months was lower for EVLTAP than for EVLT alone (0 (0-1) versus 2 (0-2); P < 0.001), with lower Aberdeen Varicose Vein Questionnaire (AVVQ) scores at 6 weeks (7.9 (4.1-10.7) versus 13.5 (10.9-18.1); P < 0.001) and 3 months (2.0 (0.4-7.7) versus 9.6 (2.2 13.8); P = 0.015). At 1 year, there were no differences in VCSS or AVVQ scores. CONCLUSION: Concomitant phlebectomy with EVLT prolonged the procedure, but reduced the need for secondary procedures and significantly improved quality of life and the severity of venous disease. PMID- 19283746 TI - Branch duct intraductal papillary mucinous neoplasms in a retrospective series of 190 patients. AB - BACKGROUND: A consensus conference has recommended close observation of branch duct intraductal papillary mucinous neoplasms (BD-IPMNs) smaller than 30 mm, without symptoms or mural nodules. This study investigated whether these recommendations could be validated in a single-centre experience of BD-IPMNs. METHODS: Some 190 patients with radiological imaging or histological findings consistent with BD-IPMN were enrolled between 1998 and 2005. Those with less than 6 months' follow-up and no histological confirmation were excluded. RESULTS: BD IPMN was diagnosed by computed tomography and pancreatography in 105 patients and pathologically in 85. Eighteen patients had adenoma, 53 borderline malignancy, five carcinoma in situ and nine invasive carcinoma. Findings associated with malignancy were the presence of radiologically suspicious features (P < 0.001) and a cyst size of at least 30 mm (P = 0.001). Had consensus guidelines been applied, 54 patients would have undergone pancreatic resection, whereas only 28 of these patients actually had a resection; 12 of the latter patients had a malignancy compared with none of the 26 patients who were treated conservatively. CONCLUSION: A simple increase in cyst size is not a reliable predictor of malignancy. Observation is recommended for patients with a BD-IPMN smaller than 30 mm showing no suspicious features on imaging. PMID- 19283747 TI - Assessment of risk of bias in randomized clinical trials in surgery. AB - BACKGROUND: Meta-analysis of randomized clinical trials (RCTs) with low risk of bias is considered the highest level of evidence available for evaluating an intervention. Bias in RCTs may overestimate or underestimate the true effectiveness of an intervention. METHODS: The causes of bias in surgical trials as described by The Cochrane Collaboration, and the methods that can be used to avoid them, are reviewed. RESULTS: Blinding is difficult in many surgical trials but careful trial design can reduce the bias risk due to lack of blinding. It is possible to conduct surgical trials with low risk of bias by using appropriate trial design. CONCLUSION: The risk of providing a treatment based on a biased effect estimate must be balanced against the difficulty of conducting trials with very low risk of bias. Better understanding of the risk of bias may result in improved trials with a closer estimate of the true effectiveness of an intervention. PMID- 19283748 TI - Hidradenitis suppurativa. AB - BACKGROUND: Hidradenitis suppurativa is a chronic, recurrent, suppurative cutaneous disease. Despite its incidence, optimal medical or surgical treatment remains unclear. This review describes the disease, ranging from pathogenesis to treatment and prognosis. METHODS: Articles were sourced from PubMed and Medline, using the MeSH terms 'hidradenitis suppurativa' and 'acne inversa'. Selection of articles was based on peer review, journal, relevance and English language. RESULTS AND CONCLUSION: On the basis of histological findings, the disease is now considered inflammatory and originating from the hair follicle; therefore, the term 'acne inversa' is favoured by some experts. The exact aetiology remains obscure but smoking seems to be a major triggering factor. Treatment should be individualized according to the site and extent of the disease. Absolute cessation of smoking is essential in the treatment of hidradenitis. Management with antibiotics or other medications may relieve early symptoms, but radical surgery may be necessary for control and to prevent recurrence. PMID- 19283749 TI - The cost of laparoscopic surgery is the price of progress. PMID- 19283750 TI - Laparoscopic common bile duct exploration using a rigid nephroscope. AB - BACKGROUND: Patients with cholelithiasis and choledocholithiasis are increasingly managed with laparoscopic bile duct exploration and cholecystectomy. Large impacted bile duct stones continue to defy laparoscopic extraction. This study explored the feasibility of laparoscopic bile duct clearance using a rigid nephroscope, which is suited to extracting large stones. METHOD: This prospective study recruited patients with large bile duct stones and a bile duct wider than 8 mm on ultrasonography. In addition to standard ports for laparoscopic cholecystectomy, a custom-made 9-mm port was introduced in the epigastrium for the rigid nephroscope, which was negotiated into the bile duct through a choledochotomy. Rigid graspers and lithotripters were introduced through the nephroscope to fragment and remove the calculi. RESULTS: Between December 2005 and September 2008, 18 patients had nephroscope-guided bile duct exploration (mean(s.d.) age 49(13.9) years, bile duct diameter 11.3(2.3) mm). Three patients had solitary stones and 15 had multiple calculi. Most of the stones were removed with graspers, but the lithotripter was required in five patients. The mean(s.d.) hospital stay was 6(2.3) days. Two patients required postoperative endoscopic retrograde cholangiopancreaticography for residual stones. CONCLUSION: The rigid nephroscope was useful for laparoscopic bile duct exploration, particularly for large impacted stones. PMID- 19283751 TI - Surgical training and working time restriction. PMID- 19283753 TI - Re: Topical haemostatic agents (Br J Surg 2008; 95: 1197-1225). PMID- 19283757 TI - Measurement and decomposition kinetics of residual hydrogen peroxide in the presence of commonly used excipients and preservatives. AB - Quantitation of residual hydrogen peroxide (H(2)O(2)) and evaluation of the impact on product stability is necessary as unwanted H(2)O(2) can potentially be introduced during the manufacturing of pharmaceuticals, biologics, and vaccines. A sensitive and convenient microplate-based method with fluorescence detection for H(2)O(2) quantitation was recently reported (Towne et al., 2004, Anal Biochem 334: 290-296). This method was found to be highly robust and reproducible, with a level of detection of 0.015 ppm and a level of quantitation of 0.025 ppm (in water). The relatively small sample requirements and amenability for automation make this assay an attractive tool for detecting residual H(2)O(2) levels. Without additional manipulation, the assay can be conducted on heterogeneous solutions with significant degree of turbidity, such as the presence of suspensions or aluminum-containing adjuvants. The quantitation of H(2)O(2) and its decomposition kinetics was also studied in presence of two common vaccine preservatives (thimerosal and phenol) and eight commonly used excipients (polyols). Over time, there is a distinct, temperature dependent decrease in H(2)O(2) recovered in thimerosal and phenol containing samples versus non preservative containing controls. Based on the half-life of spiked H(2)O(2), the decay rates in eight polyols tested were found to be: ribose > sucrose > (glycerol, glucose, lactose, mannitol, sorbitol, and xylose). PMID- 19283758 TI - Partly-PEGylated Poly-L-lysine dendrimers have reduced plasma stability and circulation times compared with fully PEGylated dendrimers. AB - The current study was performed to examine the effect of 50% surface PEGylation on the plasma circulation, biodistribution and metabolism of intravenously administered Generation 4 poly-L-lysine dendrimers in rats. Partial surface PEGylation blocked the rapid and extensive vascular binding previously described for the non-PEGylated dendrimer and slowed metabolism of the core, however the effect was less than that observed for total surface coverage of PEG. Capping the residual surface amines with acetyl groups further improved plasma stability. PMID- 19283759 TI - Assessment of dermal absorption by thermogravimetric analysis: Development of a diffusion model based on Fick's second law. AB - Recent work in our laboratory shows that it is possible to reproducibly measure the weight change over time of a piece of porcine skin exposed to a chemical vapor by thermogravimetric analysis (TGA), that is, using a microbalance maintained at constant temperature and humidity. Here, we develop a diffusion model by which the TGA skin weight curve can be interpreted. A TGA-specific analytical solution for Fick's second law of diffusion was derived. The solution contains two chemical-dependent parameters; a diffusion coefficient (D) and a skin/air partition coefficient (P). The resulting function was fitted to weight curves from 83 experiments with 4 chemicals; n-butyl acetate, methanol, 2 propanol, and toluene. In most experiments, a single weight function could not adequately describe the shape of the weight curve, therefore up to five parallel compartments, each with its parameter set Di and Pi, were tested. A two compartment model was adequate for 58% of the experiments according to the Akaike Information Criterion. Dermal penetration of the chemicals was also measured with Franz diffusion cells. The diffusion coefficients obtained by the two methods agreed well. PMID- 19283760 TI - Preparation of pH-sensitive beads for NSAID using three-component gel systems. AB - The aim of this study is to prepare novel pH-sensitive beads to obtain a gastric mucosa protective formulation and to ensure drug delivery into the intestine. Diclofenac sodium was used as a model drug. Bead formation was achieved by ionotropic gelation method using three-component gel system containing sodium alginate (Na-Alg), hydroxyethylcellulose (HEC) and hydroxypropylmethylcellulose (HPMC). Factors influencing the characteristics of beads (exposure time, cross linking agent concentration, polymer ratio) were investigated by swelling and erosion tests based on gravimetric method. Drug release was tested in distilled water and/or artificial digestive fluids and evaluated with Korsmeyer-Peppas equation and Baker-Lonsdale model. The encapsulation behaviour was qualitatively indicated by differential scanning calorimetry (DSC) method. In vivo experiments were conducted to test ulcerogenicity and intestinal absorption in rats. HPMC increased the encapsulation efficiency (EE) and HEC improved the drug release in the intestinal fluids. The equilibrium water uptake (EWU) was correlated with exposure time, calcium chloride concentration and HEC amounts. Bead erosion increased proportionately to exposure time, while it reduced when calcium chloride concentrations were increased. Higher amounts of HEC increased, while higher pH values reduced the encapsulation efficacy. The in vivo experiments demonstrated that the studied encapsulation technology markedly reduced the ulcerogenic effect of diclofenac. PMID- 19283761 TI - Endobronchial ultrasound in pediatric pulmonology. AB - Endobronchial ultrasound (EBUS) is a recently introduced technique that has significantly advanced bronchoscopic techniques in adult medicine. Use of ultrasound allows far more accurate localization and sampling of both peripheral pulmonary, as well as mediastinal and hilar lesions. This has led to greater diagnostic success, with a reduced rate of complications. Its performance characteristics in adult populations are equivalent to surgical procedures previously considered gold standard, but it has dramatically reduced morbidity and mortality among patients requiring invasive diagnostic procedures, when compared to surgical approaches. We describe the types of EBUS in clinical use, the method of use, the clinical indications for each procedure, and the potential role for EBUS in pediatric pulmonology. Radial probe EBUS is used in the investigation of peripheral lung lesions and could be adopted in children to achieve accurate biopsy of such lesions. Linear probe EBUS allows minimally invasive biopsy of mediastinal and hilar lesions. It has potentially greater performance characteristics than current biopsy techniques, with no significant complications reported to date. It may be useful in the diagnosis of lymphoma, or neurogenic tumors, as well as many other diseases resulting in mediastinal or hilar lymphadenopathy. EBUS is a minimally invasive technique that allows tissue sampling of peripheral lung lesions, or mediastinal/hilar masses, with a high diagnostic accuracy, and a significantly lower morbidity and mortality than alternative approaches. The indications for and the use of EBUS in pediatric patients is certain to increase in the future. PMID- 19283762 TI - Phrenic nerve palsy in children associated with confirmed intrathoracic tuberculosis: diagnosis and clinical course. AB - In this descriptive retrospective cases series of eight cases phrenic nerve palsy in children caused by tuberculosis lymph gland infiltration of the phrenic nerve. The lymph gland enlargement was in all cases caused by culture confirmed Mycobacterium tuberculosis. The phrenic nerve palsy was on the left side in all eight cases with the presenting feature a raised diaphragm on chest radiography that was accompanied by consolidation of the left upper lobe (88%) The diagnosis of phrenic nerve palsy was confirmed by fluoroscopy of the chest. On computer tomography the outstanding features were left sided hilar and paratracheal lymph gland enlargement with displacement of the mediastinum to the right. Mediastinal displacement lead to anterior displacement of the descending aorta, which further compressed the left main bronchus. Two children had accompanying respiratory failure requiring assisted ventilation and in two additional cases the airway compression was so severe that glandular enucleation of the enlarged glands was indicated. Of the eight children five remained symptomatic after completion of TB treatment to which steroids were added for the initial month. Diaphragmatic plication was indicated in all five cases. On clinical follow-up two children had repeated respiratory tract infections secondary to underlying lung damage while the other six remained asymptomatic. PMID- 19283763 TI - Position and ventilatory response to added dead space in prematurely born infants. AB - OBJECTIVES: Prematurely born infants are at increased risk of sudden infant death syndrome (SIDS) if slept prone. WORKING HYPOTHESIS: Prematurely born infants would have an impaired response to an added dead space and lower respiratory muscle strength in the prone compared to the supine position. STUDY DESIGN: Prospective study. PATIENT-SUBJECT SELECTION: Twenty-five infants, median gestational age of 30 (range 26-32) weeks. METHODOLOGY: The infants were studied supine and prone at a median of 36 weeks postmenstrual age. Breath by breath minute volume was measured at baseline and after a dead space was incorporated into the breathing circuit; the time constant of the response was calculated. The maximum inspiratory occlusion pressure generated (MIOP) and the pressure generated over the first 100 msec (P(0.1)) during airway occlusion were assessed. RESULTS: The median time constant was longer (26 (range 8-106) sec vs. 22 (range 6-92) sec (P = 0.045)) and the median MIOP (P = 0.001) and P(0.1) (P = 0.003) were lower in the prone compared to the supine position. CONCLUSION: Prematurely born infants have a dampened response to tube breathing and reduced respiratory muscle strength in the prone compared to the supine position, which may contribute to their increased vulnerability to SIDS in the prone position. PMID- 19283764 TI - Antibody response to Pseudomonas aeruginosa in children with cystic fibrosis. AB - Cystic fibrosis (CF) is the most frequent life threatening autosomal recessive disease in white subjects. The primary cause of morbidity and mortality in children with CF is chronic pulmonary infection, mainly caused by Pseudomonas aeruginosa. The purpose of this study was to assess the value of the measurement of antibodies to P. aeruginosa in diagnosing lung infection by the bacteria in CF patients. We assessed P. aeruginosa antibody titers in CF patients from Rio de Janeiro, Brazil, using cell lysate antigens as well as recombinant PcrV, a Type III Secretion System protein. Sputum (more than 70% of the specimens) or oropharyngeal swabs were obtained whenever patients were regularly followed for their pulmonary disease. Blood samples were obtained with an average interval of 6 months for a period of 2 years. The ELISA cut-offs were assigned as the positive 95% confidence interval of the mean antibody levels from non-fibrocystic controls. Our data showed that most CF patients (81%) of whom were not chronically infected by P. aeruginosa (Groups I and II), had their first serology positive for rPcrV. Cell-lysate ELISA was able to detect P. aeruginosa antibodies before positive culture in the first serum sample of 44% of the patients from Groups I and II. When serum reactivity to rPcrV and cell lysate were combined, 94% of CF patients from Groups I and II (n = 16) had the first serology positive for P. aeruginosa over a mean time of 20 months before the first isolation of P. aeruginosa. In conclusion, longitudinal P. aeruginosa serology should become part of respiratory care follow-up, in conjunction with other lung parameter functions. PMID- 19283765 TI - A photophysical study on the role of bile salt hydrophobicity in solubilizing amphotericin B aggregates. AB - Amphotericin B (AmB) is a highly effective antifungal agent and finds utility against a broad spectrum of fungal species. Bile salts are biocompatible biosurfactants, widely used as drug delivery media for many hydrophobic drugs. AmB in the colloidal suspension of sodium deoxycholate (NaDC) is a well-known commercial formulation of AmB. In the present work, the association of AmB with three bile salts, namely sodium cholate, sodium taurodeoxycholate and sodium taurocholate is studied using the photophysical properties of AmB. Selective excitation of monomeric AmB (lambda(ex) 414 nm, lambda(em) 560 nm) and dimeric AmB (lambda(ex) 335 nm, lambda(em) 472 nm) reveal that with increasing concentration of bile salts, the higher aggregates in water disaggregate to form both monomeric and dimeric forms of AmB. This is seen to be a general trend in all the bile salts studied. Results of steady state fluorescence anisotropy and fluorescence lifetimes studies suggest that the interaction between AmB (hydrophobic heptaene face) and bile salts (hydrophobic steroidal face) is essentially hydrophobic. PMID- 19283766 TI - Correlation between in vitro dissolution profiles from enteric-coated dosage forms and in vivo absorption in rats for high-solubility and high-permeability model drugs. AB - We examined the in vitro dissolution-in vivo absorption correlation (IVIVC) for enteric-coated granules containing theophylline, antipyrine or acetaminophen as model drugs with high solubility and high permeability. More than 85% of each drug was released from granules coated with hypromellose acetate succinate (HPMCAS) (AS-LG grade, which dissolves at pH above 5.5) at a mean dissolution rate of more than 5 %/min after a lag time of less than 4 min in simulated intestinal fluid of pH 6.8. The lag time and the dissolution rate were significantly extended and reduced, respectively, when AS-LG was replaced with AS HG (a grade of HPMCAS that dissolves at pH above 6.8). Enteric-coated granules were administered intraduodenally to anesthetized rats. Statistical significances of differences of in vitro lag time between AS-LG- and AS-HG-coated granules were consistent with those in vivo, for all drugs. Significant differences in dissolution rates between granules also corresponded to those in absorption rates calculated using a deconvolution method, and both parameters had comparable absolute values, except in the case of antipyrine-containing granules with relatively fast dissolution rates. Thus, a good IVIVC was generally obtained; however, the exception suggests the importance of developing a dissolution test that fully reflects the in vivo situation. PMID- 19283767 TI - Novel methods for the assessment of miscibility of amorphous drug-polymer dispersions. AB - A typical approach to miscibility analysis of amorphous drug-excipient dispersions involves measuring the glass transition temperature, T(g), using differential scanning calorimetery (DSC). Recently, we discussed two computational methods for the miscibility analysis of amorphous dispersions using X-ray powder diffraction (XRPD). Those methods could be used to qualify an amorphous dispersion as miscible or phase separated, with the implication that miscible dispersions are more stable towards recrystallization. The methods were limited by the need for reference XRPD patterns of both the amorphous drug and excipient. In this work, we propose two additional computational approaches that overcome that limitation and can be used to quantify the degree of miscibility in an amorphous dispersion. The first approach is based on the use of a Pure Curve Resolution Method to extract unknown amorphous references as well as qualify miscibility. The second method, based on Alternate Least Squares, can then be used to quantify the degree of miscibility by determining the nearest neighbor (NN) coordination number for the active pharmaceutical ingredient (API) and excipient. It is proposed that the NN coordination number is related to physical stability. PMID- 19283768 TI - Fabrication via a nonaqueous nanoprecipitation method, characterization and in vitro biological behavior of N(6)-cyclopentyladenosine-loaded nanoparticles. AB - A novel nonaqueous nanoprecipitation method was proposed to achieve the encapsulation of a small weight hydrophilic drug (N(6)-cyclopentyladenosine, CPA) in PLGA nanoparticles using a mixture of cottonseed oil and Tween-80 as nonsolvent phase. The nanoparticles were characterized in vitro as concerns size, morphology, drug loading, drug release, and drug stability in human blood. Human retinal pigment epithelium (HRPE) cells were employed to study intracellular accumulation of encapsulated or free CPA with and without unloaded particles, in the presence or absence of an equilibrative nucleoside transporter inhibitor. The particles displayed a mean size lower than 300 nm and a drug loading considerably higher than that found by conventional encapsulation methods. The suitable in vitro release properties permitted to obtain good drug stabilization in the blood. Studies on HRPE cells suggested that CPA can permeate their membrane by both diffusive- and transport-mediated mechanisms. The loaded and unloaded nanoparticles appeared able to increase the permeation rate of the diffusive mechanism, without interfering with the transporter. PMID- 19283769 TI - Investigation of the micellar effect of pluronic P85 on P-glycoprotein inhibition: cell accumulation and equilibrium dialysis studies. AB - The objective of this study was: (1) to characterize the P-gp inhibitory effect of different concentrations of Pluronic P85 on anti-HIV-1 drug cellular accumulation, and (2) to investigate the relationship between cellular accumulation and free fraction of drug. Cellular accumulation studies in MDCKII WT and MDCKII-MDR1 cell monolayers showed a biphasic dose response characterized by decline in accumulation at Pluronic concentrations greater than the CMC. This phenomenon was independent of the inhibition of P-gp efflux by Pluronic. Cell free equilibrium dialysis was used to determine the effect of Pluronic P85 on drug free fraction and the affinity of Pluronic micelles for drug was modeled. Nelfinavir and saquinavir associated extensively with micelles and equilibrium free fractions were low at P85 concentrations above the CMC, with association constants being in the order nelfinavir > saquinavir >>> abacavir. Abacavir, a P gp substrate, showed no association with micelles yet showed a biphasic response in cellular accumulation. These data suggest that, above the CMC, inhibition of P gp is not affected but rather factors such as micellar trapping could contribute to decreased accumulation. Therefore, the in vitro evaluation of the effect of Pluronic formulations on active transport should take into account both the physicochemical properties of drug and the composition of Pluronic. PMID- 19283770 TI - Synthesis and evaluation of N-nicotinoyl-2-{2-(2-methyl-5-nitroimidazol-1 yl)ethyloxy}-D,L-glycine as a colon-specific prodrug of metronidazole. AB - Metronidazole (MTZ) is a drug of choice for protozoal infections such as luminal amoebiasis. We designed and synthesized N-nicotinoyl-2-{2-(2-methyl-5 nitroimidazol-1-yl)ethyloxy}-D,L-glycine (NMG) as a colon-specific prodrug of MTZ. The synthetic yield of NMG was about 34%. The apparent partition coefficient of MTZ was greatly reduced by the chemical modification. While (bio)chemically stable in the contents of the upper intestine, NMG was rapidly cleaved to liberate MTZ on incubation with the cecal contents of rats. MTZ metabolized quickly in the cecal contents at least partly by a microbial nitroreductase, suggesting that the metabolism of MTZ is relevant to its bioactivation leading to amoebicidal action. The systemic absorption, analyzed by the blood concentration and urinary recovery of NMG, was very low after oral administration of NMG. In parallel with this, whereas MTZ disappeared mostly during the transit of the proximal small intestine, a substantial amount of NMG remained in the small intestine moving down to the large intestine where it metabolized rapidly. Moreover, comparing systemic absorption of MTZ after oral administration of NMG or MTZ, NMG markedly reduced the systemic absorption. These results suggest that NMG is a potential colon-specific prodrug of MTZ which improves therapeutic and toxicological properties. PMID- 19283771 TI - Oxidative demethylenation and subsequent glucuronidation are the major metabolic pathways of berberine in rats. AB - Present study was designed to explore roles of metabolic clearance in the disposition of berberine (BBR) in rats, with a focus on oxidative metabolism and subsequent glucuronidation. Plasma from rats after intravenous administration of BBR was collected to identify and quantify BBR and its major metabolites. The major circulating metabolites of BBR were oxidative metabolites M1 (via demethylation) and M2 (via demethylenation) and their corresponding glucuronides, with M2-glucuronide approximately 24-fold higher than M1-glucuronide. Incubations with rat liver microsomes were conducted to examine formation kinetics of two oxidative metabolites-M1 and M2, and depletion kinetics of M1 and M2, leading to the formation of glucuronide conjugates. Efforts were also made to examine roles of key CYPs and UGTs isoforms responsible for BBR metabolism using known chemical inhibitors and/or substrates. In vitro, the formation of M1 and M2 were comparable and multiple CYP enzymes were involved. In contrast, the glucuronidation of M2 was much faster than that of M1. Inhibition studies using well-characterized UGT substrates suggested both M1 and M2 could be glucuronidated by UGT1A1 and UGT2B1 while M2 glucuronidation was favored by UGT1A1. In summary, oxidative demethylenation and the subsequent glucuronidation were the major metabolic pathways of BBR in rats. PMID- 19283772 TI - Modeling the solubility of pharmaceuticals in pure solvents and solvent mixtures for drug process design. AB - The knowledge of the solubility of pharmaceuticals in pure solvents and solvent mixtures is crucial for designing the crystallization process of drug substances. The first step in finding optimal crystallization conditions is usually a solvent screening. Since experiments are very time consuming, a model which allows for solubility predictions in pure solvents and solvent mixtures based only on a small amount of experimental data is required. In this work, we investigated the applicability of the thermodynamic model perturbed-chain statistical associating fluid theory (PC-SAFT) to correlate and to predict the solubility of exemplary five typical drug substances and intermediates (paracetamol, ibuprofen, sulfadiazine, p-hydroxyphenylacetic acid, and p-aminophenylacetic acid) in pure solvents and solvent mixtures. PMID- 19283773 TI - Characterization of high-molecular-weight nonnative aggregates and aggregation kinetics by size exclusion chromatography with inline multi-angle laser light scattering. AB - Size exclusion chromatography with an inline multi-angle light scattering detector (SEC-MALS) was assessed as a means to characterize and monitor the formation of soluble, high-molecular-weight (HMW) protein aggregates so as to better quantify and model nonnative aggregation kinetics. Assay configuration and robustness were tested with respect to sample preparation, column type, and assay variability. Independent comparison of SEC-MALS with batch light scattering analysis indicates good agreement between the two methods. Weight-average molecular weight (M(w)), radius of gyration (R(g)), apparent polydispersity, and mass fraction monomer (m) together are shown to provide qualitative and quantitative experimental signatures to distinguish high-MW aggregate growth via chain polymerization versus that via aggregate-aggregate condensation. Mechanistic treatment of aggregation kinetics monitored by SEC-MALS is illustrated by data regression using a recently developed Lumry-Eyring Nucleated Polymerization model that explicitly accounts for aggregate nucleation and competing growth via chain- and condensation-polymerization. The combination of time-dependent M(w) and m data are shown to provide a convenient and robust means to separate and quantify characteristic time scales or rate coefficients for concurrent stages of irreversible aggregation. In addition, the scaling of R(g) with M(w) for HMW aggregates provides useful insights into aggregate morphology and mechanisms of aggregate growth. PMID- 19283774 TI - Ziprasidone as an adjuvant for clozapine- or olanzapine-associated medical morbidity in chronic schizophrenia. AB - OBJECTIVE: This study sought to examine the effect of ziprasidone on olanzapine or clozapine-associated medical morbidity such as insulin resistance, diabetes mellitus (DM) and impaired fasting glucose, obesity, and hyperlipidemia in patients with schizophrenia or schizoaffective disorder. METHOD: This was a 6 week, open label trial of ziprasidone 160 mg/day added to a stable dose of olanzapine or clozapine in 21 schizophrenia or schizoaffective patients with DM, impaired fasting glucose, or insulin resistance. RESULTS: Ten olanzapine-treated subjects and 11 clozapine-treated subjects were enrolled in the study. There were no significant differences between the two groups at baseline for age, gender, education, ethnicity, BMI, cholesterol levels, or fasting glucose. At week 6, there were no significant changes in weight, BMI, cholesterol levels, or fasting glucose. There was no significant difference in psychotic, negative, or depressive symptoms. QTc significantly increased at week 2 but not at week 6. CONCLUSIONS: The addition of 160 mg/day of ziprasidone was well tolerated but did not produce significant improvement in fasting glucose, insulin resistance, hyperlipidemia or lead to weight loss in olanzapine- or clozapine-treated subjects with schizophrenia or schizoaffective disorder. PMID- 19283775 TI - Life-threatening retroperitoneal bleeding due to warfarin-drug interactions. AB - Warfarin-drug interactioions, which can result in life-threatening bleeding, are preventable. A 53-year-old man was admitted to hospital with exacerbation of chronic obstructive airways disease, cor pulmonale, pneumonia, deep vein thrombosis and acute pulmonary embolism. His pulmonary thromboemobolism was initially treated by low-molecular-weight-heparin and heparin. After a loading dose of 5 mg for 2 days, warfarin was given in a daily dose of 2 mg. On the fifth day of warfarin therapy, the last dose of Enoxaparin was given in the morning. He had a fall in the bathroom with blunt injury to the right flank. He complained of right thigh numbness and increasing pain and swelling over his right flank and abdomen. A tender mass was noted over the right flank. His Hb level dropped to 9.7 g/dl. His INR increased from 2.46 to 3.49-3.71 one day later. On further questioning, he admitted self applications of 20 g of Analgesic balm (50% methyl salicylate) over his right calf for 3 days. CT scan showed a large right retroperitoneal haematoma and right iliacus intramuscular haematoma. Warfarin was withheld. He was given fresh frozen plasma, packed cells and vitamin K(1). Inferior vena cava filter was inserted. The haematomas were resolving. He was subsequently discharged to convalescence hospital for continuation of anticoagulant therapy and close monitoring. Significant usage of topical methyl salicylate ointment can potentiate the anticoagulant effect of warfarin. The over anticoagulation and the presence of platelet dysfunction increase the risk of severe bleeding, which can be provoked by trauma. PMID- 19283776 TI - Neuronal circuit-dependent alterations in expression of two isoforms of glutamic acid decarboxylase in the hippocampus following electroconvulsive shock: A stereology-based study. AB - There is an increasing body of evidence suggesting that GABAergic dysfunction is involved in various psychiatric disorders. The goal of our study was to investigate the influences of electroconvulsive therapy (ECT), one of the most effective treatments for depression, on the GABAergic system in the hippocampus. In this stereology-based study, we identified GABAergic neurons by immunostaining for two isoforms of glutamic acid decarboxylase (GAD), GAD65, and GAD67 and estimated the expression changes induced by single or repeated electroconvulsive shock (ECS; an animal model of ECT). The numerical density (ND) of entire population of GABAergic neurons (expressing GAD65 and/or GAD67) was seldom altered by the administration of ECS. GAD67-positive (GAD67(+)) neurons were also rarely affected by ECS. On the other hand, the ND of GAD65(+) neurons was changed in a layer-specific manner. In the CA1 region, the ND of GAD65(+) neurons was increased in the strata radiatum/lacunosum-moleculare (SR/SLM) by repeated ECS. In the CA3 region, the ND of GAD65(+) neurons was decreased in the stratum oriens and SR/SLM after single ECS. The expression ratio of GAD65 in GABAergic neurons was increased specifically in layers receiving afferents from the entorhinal cortex (EC), i.e., SR/SLM of the CA1 region and molecular layer of the dentate gyrus (DG), after repeated ECS administration, whereas the expression ratio of GAD67 in GABAergic neurons was decreased in several layers by the same treatment. These results indicate that the ECS-induced changes in ND of GAD65(+) or GAD67(+) neurons were most likely due to alterations in GAD expression rather than actual increases or decreases in cell numbers. Altogether, the neuronal circuit dependent alterations in GABA-mediated signaling may play a contributory role in the depression treatment process introduced by ECT. PMID- 19283777 TI - T-cell tolerance induced by repeated antigen stimulation: selective loss of Foxp3 conventional CD4 T cells and induction of CD4 T-cell anergy. AB - Repeated immunization of mice with bacterial superantigens induces extensive deletion and anergy of reactive CD4 T cells. Here we report that the in vitro proliferation anergy of CD4 T cells from TCR transgenic mice immunized three times with staphylococcal enterotoxin B (SEB) (3 x SEB) is partially due to an increased frequency of Foxp3(+) CD4 T cells. Importantly, reduced number of conventional CD25(-) Foxp3(-) cells, rather than conversion of such cells to Foxp3(+) cells, was the cause of that increase and was also seen in mice repeatedly immunized with OVA (3 x OVA) and OVA-peptide (OVAp) (3 x OVAp). Cell transfer experiments revealed profound but transient anergy of CD4 T cells isolated from 3 x OVAp and 3x SEB mice. However, the in vivo anergy was CD4 T cell autonomous and independent of Foxp3(+) Treg. Finally, proliferation of transferred CD4 T cells was inhibited in repeatedly immunized mice but inhibition was lost when transfer was delayed, despite the maintenance of elevated frequency of Foxp3(+) cells. These data provide important implications for Foxp3(+) cell mediated tolerance in situations of repeated antigen exposure such as human persistent infections. PMID- 19283778 TI - The IFN regulatory factor 7-dependent type I IFN response is not essential for early resistance against murine cytomegalovirus infection. AB - IFN regulatory factor 7 (IRF7) has been described as the master regulator of type I IFN responses and has been shown to be critical for innate antiviral immunity in vivo. In addition to type I IFN, NK cell responses are involved in the control of viral replication during acute viral infection. To investigate the role of IRF7 in the context of a viral infection that induces a strong NK cell response, the murine cytomegalovirus (MCMV) infection model was used. WT, IRF7-deficient and IRF3/IRF7-double deficient mice were infected with MCMV. The systemic IFN alpha response to MCMV was entirely dependent on IRF7, but independent of IRF3. However, peak IFN-beta production during MCMV infection was not affected by the lack of IRF7 or both IRF7 and IRF3. Despite the complete lack of IFN-alpha production IRF7- and IRF3/IRF7-deficient mice were surprisingly efficient in controlling MCMV replication and were only modestly more susceptible to MCMV infection than WT mice. NK cell cytotoxicity was unimpaired and NK cell IFN-gamma production was enhanced in IRF7-deficient mice correlating with increased levels of bioactive IL-12. Owing to these compensatory mechanisms IRF7-dependent antiviral immune responses were not essential for resistance against acute MCMV infection in vivo. PMID- 19283779 TI - Dynamic regulation of the P2X4 receptor in alveolar macrophages by phagocytosis and classical activation. AB - ATP-gated P2X(4) receptors (P2X(4)R) in macrophages and microglia have been implicated in neuropathic and inflammatory pain by currently unidentified mechanisms. P2X(4)R are found predominantly in intracellular lysosomal compartments but can be rapidly trafficked to the surface membrane by procedures that induce endolysosomal secretion. We studied total and surface membrane P2X(4)R protein expression by Western blot and biotinylation assays and functional expression by whole-cell patch clamp assays in human and rat alveolar macrophages in response to phagocytosis of zymosan and opsonized zymosan bioparticles and to classical and alternative macrophage activation. Unstimulated macrophages showed high total protein expression but very low functional expression. Phagocytosis rapidly (within 4 h) increased functional P2X(4)R expression by 2- to 7-fold as did chloroquine, an agent known to induce lysosomal secretion. In contrast, classical activation of macrophage for 48 h with IFN gamma and TNF-alpha or IFN-gamma and LPS reduced surface and functional P2X(4)R expression by 3-fold without altering total P2X(4)R protein levels. Alternative activation with IL-4 or IL-13 did not alter total, surface or functional expression of P2X(4)R. This is the first study of the regulation of P2X(4)R in macrophages by physiological stimuli and presents a picture whereby P2X(4)R become functional in response to initial phagocytic stimuli but return to a non functional state during sustained activation by classical macrophage activation. PMID- 19283780 TI - Loss of FOXP3 expression in natural human CD4+CD25+ regulatory T cells upon repetitive in vitro stimulation. AB - The adoptive transfer of CD4(+)CD25(+) natural regulatory T cells (Treg) is a promising strategy for the treatment of autoimmune diseases and the prevention of alloresponses after transplantation. Clinical trials exploring this strategy require efficient in vitro expansion of this rare cell population. Protocols developed thus far rely on high-grade purification of Treg prior to culture initiation, a process still hampered by the lack of Treg cell-specific surface markers. Depletion of CD127(+) cells was shown to separate activated conventional T cells from natural Treg cell populations allowing the isolation of highly enriched FOXP3(+) cells with all functional and molecular characteristics of natural Treg. Here, we demonstrate that upon in vitro expansion, CpG methylation in a conserved region within the FOXP3 gene locus increased in CD4(+)CD25(+)CD127(low) Treg, correlating with loss of FOXP3 expression and emergence of pro-inflammatory cytokines. Further analysis identified CD45RA( )FOXP3(+) memory-type Treg as the main source of converting cells, whereas CD45RA(+)FOXP3(+) Treg from the same donors showed no conversion within 3 wk of in vitro expansion. Thus, Treg cell lineage differentiation does not seem to represent a final fate decision, as natural Treg can lose their cell-type specific characteristics after repetitive TCR stimulation. PMID- 19283781 TI - Thymus-specific serine protease regulates positive selection of a subset of CD4+ thymocytes. AB - Thymus-specific serine protease (TSSP) was initially reported as a putative protease specifically expressed in the endosomal compartment of cortical thymic epithelial cells (cTEC). As such, TSSP is potentially involved in the presentation of the self-peptides that are bound to MHC class II molecules expressed at the cTEC surface and are involved in the positive selection of CD4(+) thymocytes. We tested this hypothesis by generating mutant mice deprived of Prss16, the gene encoding TSSP. TSSP-deficient mice produced normal numbers of T cells, despite a decrease in the percentage of cTEC expressing high surface levels of MHC class II. By using sensitive transgenic models expressing MHC class II-restricted TCR transgenes (Marilyn and OT-II), we showed that the absence of TSSP markedly impaired the selection of Marilyn and OT-II CD4(+) T cells. In contrast, selection of CD8(+) T cells expressing an MHC class I-restricted TCR transgene (OT-I) was unaffected. Therefore, TSSP is involved in the positive selection of some CD4(+) T lymphocytes and likely constitutes the first serine protease to play a function in the intrathymic presentation of self-peptides bound to MHC class II complexes. PMID- 19283782 TI - New assay to detect low-affinity interactions and characterization of leukocyte receptors for collagen including leukocyte-associated Ig-like receptor-1 (LAIR 1). AB - Leukocyte activity is controlled by numerous interactions between membrane receptors and ligands on the cell surface. These interactions are of low affinity making detection difficult. We developed a sensitive assay that could readily detect extremely weak interactions such as that between CD200 and the activating receptor CD200RLa (K(d)>500 microM) at the protein level. We used the new technology to screen for interactions of inhibitory receptors for collagens. We confirmed that both human and mouse leukocyte-associated Ig-like receptor-1, and in addition the related inhibitory leukocyte Ig-like receptor subfamily B member 4 (CD85K, Gp49B), bound collagen specifically, whereas other cell surface proteins gave no binding. The monomeric affinities of the interactions were then determined to allow comparison with other leukocyte interactions and indicate conditions when these interactions might lead to inhibitory signals. PMID- 19283783 TI - Practical guidelines for characterization of O-diglycosyl flavonoid isomers by triple quadrupole MS and their applications for identification of some fruit juices flavonoids. AB - Fifteen flavonoid O-diglycosides with different interglycosidic linkage isomery and glycosylation position have been studied in order to analyze their fragmentation patterns. Initial separation was carried out using high performance liquid chromatography with diode array detection (HPLC/DAD) coupled to an electrospray ionization (ESI) interface and a triple quadrupole mass spectrometer. Some useful differences in their MS spectra have been found and discussed. As it has already been reported, [Y*]+/[Y0]+ ratio for flavanones and [Y1]+/[Y0]+ ratio for other flavonoids is specific for each isomeric interglycosidic linkage. In this work it has also been observed that the abundance of these ions is dependent on the position of glycosylation. On the basis of these differences, systematic guidelines for our experimental conditions have been proposed for the differentiation of not only isomeric interglycosidic linkage but also glycosylation position using collision-induced dissociation MS/MS (CID-MS/MS) spectra in positive mode. These results have been successfully applied for the characterization of three diglycosyl flavonoids found in Citrus fruit juices and these conclusions have also been extrapolated for characterizing two triglycosides in the same fruits. PMID- 19283784 TI - A mass spectrometry based imaging method developed for the intracellular detection of HIV protease inhibitors. AB - Mass spectrometry imaging is a promising technique for measuring drugs and drug metabolites in cells and tissues. In this manuscript we describe a method for the imaging of HIV protease inhibitors. As a model system we used Mono Mac 6 cells cultured with the HIV protease inhibitors saquinavir and nelfinavir deposited on glass slides using a cytocentrifuge. A sublimation/deposition device for homogeneous matrix deposition was constructed which allows imaging of these HIV protease inhibitors at clinically relevant concentrations. Using this matrix sublimation/deposition method, glass slides containing the cytocentrifuged cells can be measured and analyzed by two types of mass spectrometry techniques, viz. matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and MALDI Fourier transform ion cyclotron resonance (FTICR), and this makes it possible to perform imaging rapidly (MALDI-TOF) and with a very high selectivity (MALDI FTICR). PMID- 19283785 TI - Electrospray ionization mass spectrometry as a critical tool for revealing new properties of snake venom phospholipase A2. AB - Results from high-performance liquid chromatography/nano-electrospray ionization tandem mass spectrometry (HPLC/nESI-MS/MS) coupled to two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2D SDS-PAGE) indicated that the monomer and dimer of phospholipase A(2) (PLA(2)) coexisted in crude Chinese Agkistrodon blomhoffii Ussurensis snake venom (ABUSV). Then, an acidic PLA(2) with the accurate molecular mass of 13979.6 Da was purified from ABUSV (mo-ABUSV aPLA(2)). MS/MS-derived peptides from ABUSV-aPLA(2) were compared with other homologous snake venom PLA(2)s, which in turn showed that ABUSV-aPLA(2) is a novel snake venom PLA(2). Meanwhile, the ABUSV-aPLA(2) dimer (di-ABUSV-aPLA(2)) was also obtained. MS/MS analysis identified the same peptides from di-ABUSV aPLA(2) as from mo-ABUSV-aPLA(2), which indicates that di-ABUSV-aPLA(2) is a homodimer. One Ca(2+) ion is contained per ABUSV-aPLA(2). The Ca(2+) ion is critical for both the hydrolytic activity and the structure of ABUSV-aPLA(2). Pro Q Emerald and Pro-Q Diamond specific glycoprotein and phosphoprotein staining combined with MS/MS analysis indicated that the ABUSV-aPLA(2) is both a glycoprotein and a phosphoprotein, which to our knowledge is the first such report for a snake venom PLA(2) and thus provides new threads for the study of the functions and structures of snake venom PLA(2)s. One phosphorylation site and the size of the glycan chain are determined by using HPLC/nESI-MS/MS and matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. The delicate utilization of ESI-MS can exert tremendous impact on protein sciences. PMID- 19283786 TI - A metabolomic analysis of the toxicity of Aconitum sp. alkaloids in rats using gas chromatography/mass spectrometry. AB - A metabolomic investigation of intoxication with Aconitum sp. alkaloids was carried out. Plasma obtained from Wistar rats administered these alkaloids was analyzed using gas chromatography/time-of-flight mass spectrometry. Samples were analyzed following protein precipitation, liquid-liquid extraction, and derivatization. Thirty-six metabolites were identified among the detected compounds. Subsequent data analysis using the t test and principal component analysis revealed metabolic differences between the control rats and treated groups as well as between the groups of rats administered different alkaloids. Twenty-seven metabolites were significantly different between plasma samples from the controls and treated groups. The significant decreased plasma levels of glutamine and creatinine in all treated groups suggested impaired heart and muscle function caused by alkaloids. The plasma levels of 22 metabolites in the hypaconitine group were significantly decreased. In contrast, only 8 and 13 metabolites were observed with significantly decreased levels in the aconitine and mesaconitine groups, respectively.These results indicated that Aconitum sp. alkaloids can cause metabolic disorders in rats. The toxicity and corresponding mechanism of hypaconitine was different from those of aconitine and mesaconitine, based on the differences of perturbed metabolic patterns between groups. PMID- 19283787 TI - Identification of hydroxyl radical oxidation products of N-hexanoyl-homoserine lactone by reversed-phase high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. AB - A reversed-phase high-performance liquid chromatography/electrospray tandem mass spectrometry method was developed for the characterization of hydroxyl radical oxidation products of N-hexanoyl-homoserine lactone (C6-HSL), a member of the N acylhomoserine lactone (AHL) class of microbial quorum-sensing signaling molecules identified in many Gram-negative strains of bacteria. Six products were identified: four with molecular weight (MW) of 213 and two with MW of 260. The characteristic product ions formed through collision-induced dissociation (CID) provided diagnostic structural information. One of the photolysis products was determined to be N-(3-oxohexanoyl)homoserine lactone (3OC6-HSL), a highly active quorum-sensing signal, by comparison with a reference standard. Three structural isomers with the same mass as 3OC6-HSL were identified as acyl side chain oxidized C6-HSL (keto/enol functionalized) by accurate mass measurement and the structures of these products were proposed from CID spectral interpretation. Two structural isomers formed from concurrent oxidation and nitration of C6-HSL were also observed and their structures were postulated based on CID spectra. In addition to the six hydroxyl radical oxidation products formed from the C6-HSL precursor, five additional compounds generated from combined oxidation and lactonolysis of C6-HSL were identified and structures were postulated. PMID- 19283788 TI - Critical assessment of the applicability of gas chromatography-combustion-isotope ratio mass spectrometry to determine amino sugar dynamics in soil. AB - Amino sugars in soils have been used as markers of microbial necromass and to determine the relative contribution of bacterial and fungal residues to soil organic matter. However, little is known about the dynamics of amino sugars in soil. This is partly because of a lack of adequate techniques to determine 'turnover rates' of amino sugars in soil. We conducted an incubation experiment where (13)C-labeled organic substrates of different quality were added to a sandy soil. The objectives were to evaluate the applicability of compound-specific stable isotope analysis via gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) for the determination of (13)C amino sugars and to demonstrate amino sugar dynamics in soil. We found total analytical errors between 0.8 and 2.6 per thousand for the delta(13)C-values of the soil amino sugars as a result of the required delta(13)C-corrections for isotopic alterations due to derivatization, isotopic fractionation and analytical conditions. Furthermore, the delta(13)C-values of internal standards in samples determined via GC-C-IRMS deviated considerably from the delta(13)C-values of the pure compounds determined via elemental analyzer IRMS (with a variation of 9 to 10 per thousand between the first and third quartile among all samples). This questions the applicability of GC-C-IRMS for soil amino sugar analysis. Liquid chromatography-combustion-IRMS (LC-C-IRMS) might be a promising alternative since derivatization, one of the main sources of error when using GC-C-IRMS, is eliminated from the procedure. The high (13)C-enrichment of the substrate allowed for the detection of very high (13)C-labels in soil amino sugars after 1 week of incubation, while no significant differences in amino sugar concentrations over time and across treatments were observed. This suggests steady-state conditions upon substrate addition, i.e. amino sugar formation equalled amino sugar decomposition. Furthermore, higher quality substrates seemed to favor the production of fungal-derived amino sugars. PMID- 19283789 TI - Effect of occupation and smoking on respiratory symptoms in working children. AB - OBJECTIVES: To compare the respiratory symptoms and the lung function of children who work in different occupational groups. METHODS: The study was performed among children attending vocational training centers. The participants were evaluated in six different occupation groups. Chronic cough, wheezing and shortness of breath were evaluated by questionnaire. The association of independent variables with the respiratory symptoms was investigated through both univariate and multivariate methods. RESULTS: Among 642 children, 534 were males; the mean age was 17.7 +/- 1.0 years. Using an internal reference group, the odds ratios of chronic cough were significantly higher in the lathe (OR: 2.0, 95%CI: 1.07-3.74), coiffure (OR: 1.94. 95%CI: 1.01-3.70), and electricity-construction (OR: 2.63, 95%CI: 1.06-6.54) groups after adjustment for smoking, age, gender, and work characteristics. There were no significant differences in spirometric values between occupational groups in either smoking or non-smoking males. In non smoking females, median values of FEV(1) (P: 0.046), PEF (P: 0.005) and FEF(25 75%) (P: 0.019) were lower in the textile compared to the coiffure group. There was no significant association between the total working time and spirometric values. There was no statistically significant relationship between the work related factors and the smoking status. CONCLUSIONS: The prevalence of chronic cough was higher in the lathe, coiffure, and electricity-construction groups and pulmonary function tests were lower in the non-smoking textile female group. Working children should be screened for respiratory symptoms and disease. PMID- 19283790 TI - Ionization efficiency of alpha-helical peptides in laser desorption/ionization mass spectrometry. PMID- 19283791 TI - A randomized trial investigating an exercise program to prevent reduction of bone mineral density and impairment of motor performance during treatment for childhood acute lymphoblastic leukemia. AB - BACKGROUND: Reduced bone mineral density (BMD), altered body composition, impaired motor performance and passive ankle dorsiflexion are side effects of acute lymphoblastic leukemia (ALL) treatment. We performed a randomized study investigating whether an exercise program could prevent these side effects. PROCEDURE: At diagnosis we randomized 51 ALL patients (median age: 5.4 years) into a group receiving a 2-year exercise program or a control group receiving standard care. BMD of total body (BMD(TB)), lumbar spine (BMD(LS)) and body composition were measured using dual energy X-ray absorptiometry, motor performance with Bayley Scales of Infant Development or Movement-ABC, and passive ankle dorsiflexion with a goniometer. The investigator was blinded to the randomization. RESULTS: Body fat increased equally during treatment in both groups. One year after cessation of therapy more rapid decline of excessive body fat was observed in the intervention group than in the controls (P = 0.01). Lean body mass, BMD(TB) and BMD(LS) of both groups decreased equally during treatment and increased equally thereafter. Both groups showed a similar decrease in passive ankle dorsiflexion and motor performance during treatment. Adherence to the intervention program varied considerably. Adherence to intervention: 11% of children exercised daily, 37% > once a week, 16% once weekly, 36% < once a week. CONCLUSIONS: The exercise program was not more beneficial than standard care in preventing reduction in BMD, motor performance and passive ankle dorsiflexion than standard care, most likely due to unsatisfactory compliance. Increased BMI and body fat in the intervention group normalized faster after cessation of chemotherapy. PMID- 19283792 TI - Pediatric duodenal cancer and biallelic mismatch repair gene mutations. AB - Gastrointestinal malignancies are extremely rare in the pediatric population, and duodenal cancers represent an even more unusual entity. Intestinal cancers in young adults and children have been observed to be associated with functional deficiencies of the mismatch repair (MMR) system causing a cancer-predisposition syndrome. We report the case of a 16-year-old female with duodenal adenocarcinoma and past history of medulloblastoma found to have a novel germline bialleleic truncating mutation (c.[949C>T]+[949C>T]) of the PMS2 gene. PMID- 19283793 TI - Larynx preservation clinical trial design: key issues and recommendations--a consensus panel summary. AB - BACKGROUND: To develop guidelines for the conduct of phase III clinical trials of larynx preservation in patients with locally advanced laryngeal and hypopharyngeal cancer. METHODS: A multidisciplinary international consensus panel developed recommendations after reviewing results from completed phase III randomized trials, meta-analyses, and published clinical reports with updates available through November 2007. The guidelines were reviewed and approved by the panel. RESULTS: The trial population should include patients with T2 or T3 laryngeal or hypopharyngeal squamous cell carcinoma not considered for partial laryngectomy and exclude those with laryngeal dysfunction or age more than 70 years. Functional assessments should include speech and swallowing. Voice should be routinely assessed with a simple, validated instrument. The primary endpoint should capture survival and function. The panel created a new endpoint: laryngo esophageal dysfunction-free survival. Events are death, local relapse, total or partial laryngectomy, tracheotomy at 2 years or later, or feeding tube at 2 years or later. Recommended secondary endpoints are overall survival, progression-free survival, locoregional control, time to tracheotomy, time to laryngectomy, time to discontinuation of feeding tube, and quality of life/patient reported outcomes. Correlative biomarker studies for near-term trials should include EGFR, ERCC-1, E-cadherin and beta-catenin, epiregulin and amphiregulin, and TP53 mutation. CONCLUSIONS: Revised trial designs in several key areas are needed to advance the study of larynx preservation. With consistent methodologies, clinical trials can more effectively evaluate and quantify the therapeutic benefit of novel treatment options for patients with locally-advanced laryngeal and hypopharyngeal cancer. PMID- 19283794 TI - Fabrication of a patterned assembly of semiconducting organic nanowires. PMID- 19283795 TI - Electrospun nanoparticle-nanofiber composites via a one-step synthesis. AB - A facile approach to synthesize and incorporate metal nanoparticles (NPs) into electrospun polymer nanofibers (NFs) wherein the electrospinning polymer acts as both a reducing agent for the metal salt precursor, as well as a protecting and templating agent for the ensuing NPs, is reported. Such a true one-step process at ambient conditions and free of organic solvents is demonstrated using a system comprising AgNO(3) and poly(ethylene oxide) (PEO) at electrospinnable molecular weights of 600, 1000, or 2000 kDa. The PEO transforms Ag(+) into AgNPs, a phenomenon that has not been previously possible at PEO molecular weights less than 20 kDa without the addition of a separate reducing agent and stabilizer or the application of heat. Results from X-ray photoelectron spectroscopy and UV-Vis absorption spectrophotometry analyses support the formation of pseudo-crown ethers in high molecular weight PEO as the mechanism in the development of NPs. The AgNPs reduce fiber diameter and enhance fiber quality (reduced beading) due to increased electrical conductivity. Interestingly, several of the NFs exhibit AgNP-localized nanochain formation and protrusion from the NF surface that can be attributed to the combined effect of applied electrical field on the polymer and the differences between the electrical conductivity and polarizability of the polymer and metal NPs. PMID- 19283796 TI - Block-copolymer vesicles as nanoreactors for enzymatic reactions. AB - The impact of the spatial confinement of polystyrene-block-poly(acrylic acid) (PS b-PAA) block copolymer (BCP) vesicles on the reactivity of encapsulated bovine pancreas trypsin is studied. Enzymes, as well as small molecules, are encapsulated with loading efficiencies up to 30% in BCP vesicles with variable internal volumes between 0.014 aL (internal vesicle diameter, d(in) = 30 nm) and 8 aL (d(in) = 250 nm), obtained by manipulating the vesicle preparation conditions. The kinetics of the trypsin-catalyzed reaction of a fluorogenic substrate inside and outside the vesicles is quantitatively estimated using fluorescence spectroscopic analyses in conjunction with the use of NaNO(2) as selective quencher for non-encapsulated fluorophores. The values of the catalytic turnover number obtained for reactions in differently sized nanoscale reactors show a significant increase (up to approximately 5x) with decreasing BCP vesicle volume, while the values of the Michaelis-Menten constant decrease. The observed increase in enzyme efficiency by two orders of magnitude compared to bulk solution is attributed to an enhanced rate of enzyme-substrate and molecule-wall collisions inside the nanosized reactors, as predicted in the literature on the basis of Monte Carlo simulations. PMID- 19283797 TI - Thermal deposition of intact tetrairon(III) single-molecule magnets in high vacuum conditions. AB - A tetrairon(III) single-molecule magnet is deposited using a thermal evaporation technique in high vacuum. The chemical integrity is demonstrated by time-of flight secondary ion mass spectrometry on a film deposited on Al foil, while superconducting quantum interference device magnetometry and alternating current susceptometry of a film deposited on a kapton substrate show magnetic properties identical to the pristine powder. High-frequency electron paramagnetic resonance spectra confirm the characteristic behavior for a system with S = 5 and a large Ising-type magnetic anisotropy. All these results indicate that the molecules are not damaged during the deposition procedure keeping intact the single-molecule magnet behavior. PMID- 19283798 TI - Phosphinoborane and sulfidoborohydride as chelating ligands in polyhydride ruthenium complexes: agostic sigma-borane versus dihydroborate coordination. AB - A question of coordination mode: Two new borane compounds are prepared. They act as bifunctional ligands as illustrated by their reaction with ruthenium polyhydrides which leads to the formation of two complexes (see scheme) displaying either a delta-agostic interaction of a eta(2)-B-H bond involving a trivalent boron atom or a dihydroborate ligation. PMID- 19283799 TI - Rational construction of 2D and 3D borromean arrayed organic crystals by hydrogen bond-directed self-assembly. AB - Borromean organic networks: The rigid and trigonal pyramidal molecule, 1,3,5 tris(4-carboxyphenyl)adamantane (TCA), self-assembles into a 2D Borromean linked network by hydrogen bonds. Different linkers (methanol, phenazine, 4,4' bipyridine, and 4,4'-azopyridine) result in more complex Borromean networks or a 3D polycatenation network. PMID- 19283800 TI - The shape of germanium clusters to come. AB - A different drummer: The existence of endohedral germanium clusters was predicted earlier by gas-phase experiments. The [Co@Ge(10)](3-) anion now synthesized is surprising, as it breaks with a long line of exclusively deltahedral structures found in the past. Instead, it has a regular pentagonal-prismatic structure (see picture; Co gray, Ge red). PMID- 19283801 TI - Are particulate noble-metal catalysts metals, metal oxides, or something in between? AB - A good model? Noble-metal particulate catalysts often require small amounts of oxygen to obtain optimal activity. However, the structure and stoichiometry of the oxidized metal clusters involved remains obscure, even almost two hundred years after their discovery. A heteropolypalladate salt (see picture; Pd yellow, O red) now offers a view of how oxygen might be incorporated into small noble metal clusters. PMID- 19283802 TI - Molecular recognition in self-assembled integrated circuits: getting smaller while under control. AB - Available in small print: Block copolymer lithography has great potential for reducing the size and fabrication time of integrated circuits. Hydrogen-bonding mediated molecular recognition in self-assembly processes can be used to produce highly ordered square arrays of block copolymers on the surface of a silicon substrate (see picture). PMID- 19283803 TI - Thiostannate tin-tin bond formation in solution: in situ generation of the mixed valent, functionalized complex [{(RSn(IV))2(mu-S)2}3Sn(III)2S6]. AB - In broad daylight: The double-decker thiostannate [(RSn(IV))(4)S(6)] (1, R = CMe(2)CH(2)COMe) condenses to form [{(RSn(IV))(2)(mu-S)(2)}(3)Sn(III)(2)S(6)] (2; see picture). This mixed-valent complex, which formally contains both Sn(III) and Sn(IV) atoms as confirmed by Mossbauer spectroscopy and DFT calculations, forms by a complicated, concerted mechanism. Additionally, 2 provides six carbonyl groups for further derivatization. PMID- 19283804 TI - Undulated multicompartment cylinders by the controlled and directed stacking of polymer micelles with a compartmentalized corona. AB - Like a bamboo rod: Long, bamboo-like undulated cylinders with distinct branch points and end groups (see picture) were obtained by the stacking of block terpolymer micelles that contain a fluorinated polybutadiene core and a compartmentalized corona of poly(4-tert-butoxystyrene) and poly(tert-butyl methacrylate). Stacking of the polymer micelles, which can be reversed, occurred when the solvent was changed from dioxane to ethanol. PMID- 19283805 TI - A mechanism of efficient G6PD inhibition by a molecular clip. AB - Triple duty: A synthetic molecular clip traps nicotinamide adenine dinucleotide phosphate (NADP(+); see picture) as well as occupying both the cofactor- and the substrate-binding site in glucose-6-phosphate (G6P) dehydrogenase. This combination of two inhibition mechanisms makes the clip highly effective and selective for this enzyme over other dehydrogenases. PMID- 19283806 TI - Dendrimeric oligo(phenylenevinylene)-extended dithieno[3,2-b:2',3'-d]phospholes synthesis, self-organization, and optical properties. AB - A boost from the branches: Incorporation of the dithieno[3,2-b:2',3'-d]phosphole system as a core in oligo(phenylenevinylene) dendrimers (an example is shown here) provides materials that exhibit energy-transfer features relaying incoming photons from the dendrons towards the core, which in turn shows enhanced emission intensity. The optical properties and self-assembly features of the dendrimers can be impacted by the terminal groups (-H, -CF(3), or -NPh(2)) employed.To establish this system as a fluorescent core in pi-conjugated dendrimers, a series of oligo(phenylenevinylene) (OPV)-extended dithieno[3,2-b:2',3'-d]phospholes has been prepared by means of a Wittig-Horner protocol with a dithienophosphole dialdehyde and appropriately functionalized phosphonates. The "zero-generation" model compounds have provided the general accessibility of OPV-functionalized dithienophospholes, and show varying emission colors covering the optical spectrum from green to red. Expansion of the synthetic strategy towards the corresponding first-generation dendrimers has provided materials that show intriguing self-organization features in case of the phenyl-terminated dendrimer, forming large one-dimensional microfibres, as well as desirable energy-transfer processes from the dendrons to the dithienophoshole core resulting in an enhanced emission intensity for the latter. The present study has revealed that the terminal end-groups of the OPV branches have significant impact on the optical features of the OPV dendrimers as a whole. PMID- 19283807 TI - Density functional studies on isomerization of prostaglandin H2 to prostacyclin catalyzed by cytochrome P450. AB - Reaction mechanisms for the isomerization of prostaglandin H(2) to prostacyclin catalyzed by cytochrome P450 are investigated by the unrestricted Becke's three parameter plus Lee-Yang-Parr density functional level of theory. The results show that the homolytic O-O bond cleavage of endoperoxide in prostaglandin H(2) is the rate-limiting step and that the isomerization proceeds through proton-coupled electron transfer. We located two reaction pathways through an Fe(IV)-porphyrin intermediate and an Fe(III)-porphyrin pi-cation radical intermediate. PMID- 19283808 TI - A bisfullerene-bis(dipyrrinato)zinc complex: electronic coupling and charge separation in an easy-to-assemble synthetic system. AB - Getting connected: The use of a bis(dipyrrinato)zinc(II)) linker allows the facile tethering of two C(60) subunits and gives rise to an electronically coupled system that allows effective charge separation following photoexcitation (see figure). PMID- 19283809 TI - A simple and versatile re-catalyzed Meyer-Schuster rearrangement of propargylic alcohols to alpha,beta-unsaturated carbonyl compounds. AB - Rhenium does the job! A readily available rhenium complex efficiently catalyzed the direct Meyer-Schuster-like rearrangement of different alkyl- and aryl substituted propargylic secondary and tertiary alcohols to the corresponding alpha,beta-unsaturated compounds, which were produced with virtually complete E stereoselectivity. The reaction proceeded under neutral conditions and no racemization of potentially enolizable stereocenters was observed. PMID- 19283810 TI - A simple catalyst for the efficient benzylation of arenes by using alcohols, ethers, styrenes, aldehydes, or ketones. AB - One catalyst fits all! One catalyst is active for a wide set of benzylating reactions (see scheme). A tandem process allows the use of aldehydes and ketones as benzylating agents.The compound [IrCp*(OTf)(2)(I(nBu))] (I(nBu)=1,3-di-n-butyl imidazolylidene) is an effective catalyst in the benzylation of arenes with different benzylating agents, such as alcohols, ethers and styrenes, representing an unprecedented highly versatile catalyst for this type of process. The same compound also catalyses a remarkable tandem process that allows the use of aldehydes and ketones as benzylating agents, through the base-free hydrogenation of C==O bonds with iPrOH and further use of the resulting primary or secondary alcohols as benzylating agents. PMID- 19283811 TI - Asymmetric Suzuki-Miyaura coupling in water with a chiral palladium catalyst supported on an amphiphilic resin. AB - You oughta use water: Broad functional-group (FG) tolerance was observed for the title coupling of aryl halides (X = Cl, Br, I) and aryl boronic acids to give biaryl compounds with up to 94% ee. The chiral imidazoindole phosphine-palladium catalyst supported on an amphiphilic polystyrene-poly(ethylene glycol) (PS-PEG) resin could be recycled readily. PMID- 19283812 TI - Matrix-free formation of gas-phase biomolecular ions by soft cluster-induced desorption. AB - All in a ball: Neutral molecular clusters consisting of a few thousand molecules can be seen as tiny snow balls; if they are thrown fast enough onto a surface, they are able to pick up biomolecules such as insulin from that surface. Since they break down and evaporate during and after the collision, bare biomolecular ions are available for mass spectrometry after such an energetic throw. PMID- 19283813 TI - Gold nanorod/Fe3O4 nanoparticle "nano-pearl-necklaces" for simultaneous targeting, dual-mode imaging, and photothermal ablation of cancer cells. AB - Gold and pearls: Multifunctional nanoparticles, each composed of a single, amine modified gold nanorod, decorated with multiple "pearls" of Fe(3)O(4) nanoparticles capped with carboxy groups, are prepared. Their effectiveness in simultaneous targeting, dual-mode imaging, and photothermal ablation of breast cancer cells is demonstrated. PMID- 19283814 TI - Rapid and tunable patterning of high purity ZnO nanoarrays without template or catalyst. AB - Arrays of endless possibility: Without template or catalyst, high-purity ZnO nanoarrays were patterned though a rapid, tunable and repeatable method (see picture). Their potential contributions in both electrical and biomedical fields were also discussed and demonstrated. PMID- 19283815 TI - An all-photonic molecular keypad lock. AB - Off and on: A molecular triad, consisting of a porphyrin linked to two different, independently addressable photochromic moieties, functions as a molecular keypad lock with all-photonic inputs and output. The porphyrin correlates the responses of the two inputs to light of different wavelengths and provides an appropriate output as fluorescence, which results only when one of eight possible input combinations has been applied (see figure). PMID- 19283816 TI - A germanium(II) cation with [1+1] coordination. AB - Breaking the molecular symmetry by protonation of germylene 1 is the key step in the synthesis of the germyliumylidene 2, which is stabilized by an intramolecular interaction with a distant imido group. PMID- 19283817 TI - Luminescent heteropolynuclear complexes of 3,5-dimethylpyrazolate [Pt2Au2M2(Me2pz)8] (M = Ag, Cu) showing the synergistic effect of three transition elements in the excited state. AB - Swap the coins! The Pt(2)Au(2), Pt(2)Au(2)Cu(2), and Pt(2)Au(2)Ag(2) complexes of 3,5-dimethylpyrazolate exhibit yellow-green, orange, and sky-blue luminescence, respectively (see figure). The emission energies of Pt(2)Au(2)M(2) complexes can be controlled by the change of the third coinage metal ions M. The Pt(2)Au(2)M(2) complexes take the cis configuration with respect to the Au(2)M(2) plane. PMID- 19283818 TI - A C-linked glycomimetic in the gas phase and in solution: synthesis and conformation of the disaccharide Manalpha(1,6)-C-ManalphaOPh. AB - The effect of carbon is subtle but sweet: The flexible C-linkage in the newly synthesised C-glycosyl mimetic, Manalpha(1,6)-C-ManalphaOPh allows OH--pi bonding, both in the gas phase and in aqueous solution. This interaction is absent in the O-linked disaccharide (see figure).The intrinsic conformational preference of a newly synthesised glycomimetic, the C-linked disaccharide Manalpha(1,6)-C-ManalphaOPh (1), has been determined in the gas phase at about 10 K by infrared ion dip spectroscopy coupled with density functional theory and ab initio calculations, and compared with its dynamical conformation in aqueous solution at 298 K by NMR spectroscopy. Comparisons are also made between these conformations and those of the corresponding O-linked disaccharide 2 in the gas phase and the C-linked disaccharide Manalpha(1,6)-C-ManalphaOMe (3) in the gas phase and in aqueous solution. The C- and O-linked disaccharides 1 and 2 present quite distinct conformational preferences in the gas phase: inter-glycosidic hydrogen bonding, seen in one of the two conformers populated in 2, is not seen in 1 which adopts a conformation (not populated in 2) with glycosidic dihedral angles (phi, psi, omega) of -72 degrees , 52 degrees and 66 degrees ; supported in part by an OH--pi hydrogen bond. This conformer is also strongly populated in an aqueous solution of 1 (and very weakly, of 3) together with a second conformer, with dihedral angles (phi, psi, omega) of about -60 degrees , 180 degrees and 60 degrees , not seen in the gas phase but by far the dominant conformer in an aqueous solution of 3. The C-disaccharide 1 was tested as a potential inhibitor, but displayed no significant inhibitory activity against Jack Bean alpha-mannosidase. PMID- 19283819 TI - Adducts between dansylated poly(propylene amine) dendrimers and anthracene clips mediated by Zn(II) ions: highly efficient photoinduced energy transfer. AB - A family of poly(propylene amine) dendrimers, decorated at their periphery with 4, 16, and 32 dansyl units and a molecular clip, composed of two anthracene sidewalls and a disulfate benzene bridging unit, show intense UV absorption and strong fluorescence in the visible region when in a CH(3)CN/CH(2)Cl(2) (1:1, v/v) solvent mixture. Both these classes of compounds are good ligands for Zn(II) ions, as demonstrated by the changes in the absorption and fluorescence spectra upon addition of metal ions. These coordinating properties have been exploited in the self-assembly of complex structures in which the interaction between a dansylated dendrimer and anthracene-functionalized clips is mediated by Zn(II) ions. The self-assembly process is reversible and the number of metal ions and molecular clips associated with each dendrimer increases with the generation number. In these adducts, an energy transfer process from the anthracene to yield the fluorescent excited state of dansyl takes place with almost unitary efficiency. PMID- 19283820 TI - Intramolecular alkynylogous mukaiyama aldol reaction starting from bicyclic alkanones tethered to alkynyl esters: formal total synthesis of (+/-)-hamigeran B. AB - tert-Butyldimethylsilyltriflate (TBSOTf)/NEt(3) treatment of alkynyl esters tethered to bicycloalkanones led to tricyclic allenoates with total diastereoselectivity for the ring junction. The allenoates result from an intramolecular alkynylogous Mukaiyama aldol reaction promoted by a TBSOTf/NEt(3) dual activation, with key intermediates of silylalkynylketene acetals. This novel methodology was illustrated by a formal total synthesis of (+/-)-hamigeran B. PMID- 19283821 TI - Extraction of uranium from aqueous solutions by using ionic liquid and supercritical carbon dioxide in conjunction. AB - Uranyl ions [UO(2)](2+) in aqueous nitric acid can be extracted into supercritical CO(2) (sc-CO(2)) by using an imidazolium-based ionic liquid with tri-n-butyl phosphate (TBP) as a complexing agent. The transfer of uranium from the ionic liquid to the supercritical fluid phase was monitored by UV/Vis spectroscopy using a high-pressure fiber-optic cell. The form of the uranyl complex extracted into the sc-CO(2) phase was identified to be [UO(2)(NO(3))(2)(TBP)(2)]. The extraction results were confirmed by fluorescence spectroscopy and by neutron activation analysis. This technique has potential applications in the field of nuclear waste management for extracting other actinides. PMID- 19283822 TI - Halogen bonding between an isoindoline nitroxide and 1,4 diiodotetrafluorobenzene: new tools and tectons for self-assembling organic spin systems. AB - Radical assembly: Halogen bonding has been observed for the first time between an isoindoline nitroxide and an iodoperfluorocarbon (see figure), which cocrystallize to form a discrete 2:1 supramolecular compound in which N--O(.)I halogen bonding is the dominant intermolecular interaction. This illustrates the potential use of halogen bonding and isoindoline nitroxide tectons for the assembly of organic spin systems.The isoindoline nitroxide 1,1,3,3 tetramethylisoindolin-2-yloxyl (TMIO) and 1,4-diiodotetrafluorobenzene readily form a discrete 2:1 complex that shows evidence of relatively strong N--O(.)I halogen bonding. This interaction was characterized in the solid state by single crystal X-ray analysis, thermal analysis, and vibrational spectroscopy (IR and Raman), backed by density functional theory calculations. EPR spectroscopy performed on a solution of TMIO in pentafluoroiodobenzene, a halogen-bonding donor, indicates that halogen bonding induces an increase in electron density at the nitroxide nitrogen nucleus and an increase in the nitroxide rotational correlation time. Our findings demonstrate the potential of utilizing halogen bonding interactions to promote the self-assembly of new isoindoline nitroxide tectons for the preparation of organic spin systems. PMID- 19283823 TI - Palladium(0)-mediated rapid methylation and fluoromethylation on carbon frameworks by reacting methyl and fluoromethyl iodide with aryl and alkenyl boronic acid esters: useful for the synthesis of [11C]CH(3)--C- and [18F]FCH2--C Containing PET tracers (PET=positron emission tomography). AB - A new synthetic methodology for the rapid methylation and fluoromethylation on aryl and alkenyl frameworks by using methyl and fluoromethyl iodide with an organoboronic acid ester has been developed under the simple and mild conditions of [Pd(2)(dba)(3)]/P(o-CH(3)C(6)H(4))(3)/K(2)CO(3) (dba= dibenzylideneacetone) in DMF at 60 degrees C for 5 min (see scheme). This boron protocol provides a firm chemical basis for the synthesis of (11)C- and (18)F-incorporated PET tracers.The rapid methylation and fluoromethylation on aryl and alkenyl carbon frameworks by reacting methyl and fluoromethyl iodide with aryl and alkenyl boronates have been studied with the focus on the realization of the synthesis of [(11)C]CH(3)- and [(18)F]FCH(2)-labeled positron emission tomography (PET) tracers. The coupling of methyl iodide and pinacol phenylboronate (40 equiv) is accomplished in >91 % yield within 5 min at 60 degrees C under the conditions of [Pd(2)(dba)(3)]/P(o CH(3)C(6)H(4))(3)/K(2)CO(3) (0.5:2:2; dba=dibenzylideneacetone) in DMF. The reaction shows a high generality and is applicable to various types of aryl and alkenyl boronates, giving the corresponding methylated derivatives in high yields (80-99 %). This reaction is also useful for the rapid incorporation of the fluoromethyl group. Thus, this boron protocol provides a firm chemical basis for the synthesis of (11)C- and (18)F-incorporated PET tracers and can be used as a complementary method for [(11)C]methylation together with our previous and ongoing processes using organotributylstannanes. PMID- 19283824 TI - Collagen scaffolds loaded with collagen-binding NGF-beta accelerate ulcer healing. AB - Studies have shown that exogenous nerve growth factor (NGF) accelerates ulcer healing, but the inefficient growth factor delivery system limits its clinical application. In this report, we found that the native human NGF-beta fused with a collagen-binding domain (CBD) could form a collagen-based NGF targeting delivery system, and the CBD-fused NGF-beta could bind to collagen membranes efficiently. Using the rabbit dermal ischemic ulcer model, we have found that this targeting delivery system maintains a higher concentration and stronger bioactivity of NGF beta on the collagen membranes by promoting peripheral nerve growth. Furthermore, it enhances the rate of ulcer healing through accelerating the re epithelialization of dermal ulcer wounds and the formation of capillary lumens within the newly formed tissue area. Thus, collagen membranes loaded with collagen-targeting human NGF-beta accelerate ulcer healing efficiently. PMID- 19283825 TI - Effects of hydrostatic pressure on leporine meniscus cell-seeded PLLA scaffolds. AB - Hydrostatic pressure (HP) is an important component of the loading environment of the knee joint. Studies with articular chondrocytes and TMJ disc fibrochondrocytes have identified certain benefits of HP for tissue engineering purposes. However, similar studies with meniscus cells are lacking. Thus, in this experiment, the effects of applying 10 MPa of HP at three different frequencies (0, 0.1, and 1 Hz) to leporine meniscus cell-seeded PLLA scaffolds were examined. HP was applied once every 3 days for 1 h for a period of 28 days. Constructs were analyzed for cellular, biochemical, and biomechanical properties. At t = 4 weeks, total collagen/scaffold was found to be significantly higher in the 10 MPa, 0 Hz group when compared with other groups. This despite the fact that the cell numbers/scaffold were found to be lower in all HP groups when compared with the culture control. Additionally, the total GAG/scaffold, instantaneous modulus, and relaxation modulus were significantly increased in the 10 MPa, 0 Hz group when compared with the culture control. In summary, this experiment provides evidence for the benefit of a 10 MPa, 0 Hz stimulus, on both biochemical and biomechanical aspects, for the purposes of meniscus tissue engineering using PLLA scaffolds. PMID- 19283826 TI - Nondestructive assessment of dentin demineralization using polarization-sensitive optical coherence tomography after exposure to fluoride and laser irradiation. AB - Previous studies have demonstrated that polarization-sensitive optical coherence tomography (PS-OCT) can be used to image natural and artificial caries in dentin. The purpose of this study was to measure nondestructively the severity of artificial caries lesions in dentin and determine the efficacy of intervention with anticaries agents including fluoride and lasers. Although several studies have assessed the utility of PS-OCT to image caries lesions in enamel and to quantify the lesion severity, only a few studies have focused on lesions in dentin. In this study, images of artificial dentin lesions on extracted human teeth were acquired with PS-OCT. Before exposure to an artificial demineralizing solution, three incisions were made on the sample surfaces using either Er:YAG, Nd:YAG (lambda = 355 nm), or TEA CO(2) lasers and selected areas were treated with topical fluoride to create six unique treatment areas for each of the three laser conditions investigated. The integrated reflectivity and depth of demineralization were calculated for each of the six areas on each sample using the PS-OCT images. Polarized light microscopy (PLM) and transverse microradiography (TMR) were used to measure lesion severity on histological thin sections for comparison. PS-OCT successfully measured the inhibition of demineralization by topical fluoride. Laser irradiation was not particularly effective in increasing or decreasing the rate of dentin demineralization. PLM and TMR corroborated those results. This study demonstrates that PS-OCT can be used to measure demineralization on dentin surfaces and determine the degree of inhibition of demineralization by anticaries agents. PMID- 19283827 TI - Collagen Vitrigel membranes for the in vitro reconstruction of separate corneal epithelial, stromal, and endothelial cell layers. AB - The goal of this study was to evaluate the potential suitability of collagen Vitrigel (CV) membrane as a substrate for the separate reconstruction of the three main cellular layers of the cornea. Limbal explants, keratocytes, and endothelial cells were cultured on transparent membranes made of type I collagen. The resulting cell sheets were evaluated using RT-PCR, in addition to light and electron microscopy. Tensile testing was also performed to examine the mechanical properties of CV. Limbal explant cultures resulted in partially stratified epithelial sheets with upregulation of the putative stem cell marker p63. Keratocytes cultured in serum on CV exhibited stellate morphology along with a marked increase in expression of corneal crystallin ALDH and keratocan, (a keratan sulphate proteoglycan: KSPG), compared to identical cultures on tissue culture plastic. Endothelial cells formed dense monolayers with uniform cell size, tight intercellular junctions, and expression of voltage-dependent anion channels VDAC2 and VDAC3, chloride channel protein CLCN2, and sodium bicarbonate transporter NBC1. Epithelial and endothelial cells exhibited adhesive structures (desmosomes and hemidesmosomes) and evidence of apical specialization (microplicae), while endothelial cells also produced a Descemet's membrane-like basal lamina. CV was found to possess ultimate tensile strengths of 6.8 +/- 1.5 MPa when hydrated and 28.6 +/- 7.0 MPa when dry. Taken together, these results indicate that CV holds promise as a substrate for corneal reconstruction. PMID- 19283828 TI - Spermatozoal RNAs: what about their functions? AB - The profound architectural changes that transform spermatids into spermatozoa result in a high degree of DNA packaging within the sperm head. However, the mature sperm chromatin that harbors imprinted genes exhibits a dual nucleoprotamine/nucleohistone structure with DNase-sensitive regions, which could be implicated in the establishment of efficient epigenetic information in the developing embryo. Despite its apparent transcriptionally inert state, the sperm nucleus contains diverse RNA populations, mRNAs, antisense and miRNAs, that have been transcribed throughout spermatogenesis. There is also an endogenous reverse transcriptase that may be activated under certain circumstances. It is now commonly accepted that sperm can deliver some RNAs to the ovocyte at fertilization. This review presents potential links between male-specific genomic imprinting, chromatin organization, and the presence of diverse RNA populations within the sperm nucleus and discusses the functional significance of these RNAs in the spermatozoon itself and in the early embryo following fertilization. Some recent data are provided, supporting the view that analyzing the profile of spermatozoal RNAs could be useful for assessment of male fertility. PMID- 19283829 TI - Influence of alternating L-/D-amino acid chiralities and disulfide bond geometry on the capacity of cysteine-containing reversible cyclic peptides to disperse carbon nanotubes. AB - Although single-walled carbon nanotubes (SWNTs) have exciting properties and potential applications, their hydrophobic nature makes them difficult to purify and manipulate. To fully realize the potential of SWNTs, strategies for the effective dispersion, separation, and organization of these materials must be devised. In this article, work involving the recent design and characterization of reversible cyclic peptides (RCPs) and RCP/SWNT composites will be described. The peptides in this work contain alternating L- and D-amino acid sequences, as well as N- and C-terminal cysteine residues (RCP-Cys) that allow for their covalent closure around the circumference of individual SWNTs. When RCPs are oxidized in the presence of SWNTs, dispersions are produced that are stable against dilution by dialysis without the formation of aggregates. The reported studies using Raman spectroscopy and UV/Vis/NIR were focused on answering the questions (1) does the chirality of the disulfide bond impact the capacity of the RCP-Cys to disperse SWNTs, and (2) is the alternating chirality of the amino acids in the RCP-Cys peptides important for SWNT dispersion. It was found that though Cys-containing RCPs are indeed able to disperse SWNTs, the chirality of the Cys residues on the N- and C- termini does not have a significant influence on the dispersed SWNT population. However, there is a large decrease in the dispersability by RCP-Cys when the alternating L/D-chiral pattern of amino acids is replaced with all L-amino acids. PMID- 19283830 TI - Revisiting "reverse hydrophobic effect": applicable only to coil mutations at the surface. AB - In a seminal paper, Pakula and Sauer (Nature, 1990, 344, 363-364) demonstrated that the increase in side-chain hydrophobicity has a reverse relationship with protein stability. We have addressed this problem with several examples of mutants that span at different locations in protein structure based on secondary structure and solvent accessibility. We confirmed that the stability change upon single coil mutation at exposed region is reversely correlated with hydrophobicity with a single exception. In addition, we found the existence of such relationship in partially buried coil mutants. The stability of exposed helical mutants is governed by conformational properties. In buried and partially buried helical and strand mutants properties reflecting hydrophobicity have direct relationship with stability, whereas an opposite relationship was obtained with entropy and flexibility. The structural analysis of partially buried/exposed mutants showed that the surrounding residues are important for the stability change upon mutation. These results provide insights to understand the general behavior for the stability of proteins upon amino acid substitutions. PMID- 19283831 TI - Effects of the nanostructure and nanoporosity on bioactive nanohydroxyapatite/reconstituted collagen by electrodeposition. AB - Hydroxyapatite (HA)/collagen composites were reported to induce bony growth. Various methods for preparing HA-based composites have been investigated as potential biomaterials for bone substitutes. However, no method can generate a thick nanoporous HA. A novel bone regenerative nanocomposite consisting of nano hydroxyapatite (HA), nano-amorphous calcium phosphate (ACP) and reconstituted collagen by electrodeposition was designed in this research. Specimens with and without nanoporosity were evaluated using electrochemical measurements, material analyses, and cell-material interactions. The results showed that reconstituted collagen/nano-(HA and ACP) illustrated a multinanoporous structure and enhanced biocompatibility. Nanocomposite was comprised to nano-(HA and ACP) and reconstituted collagen. The core cell structure was formed during electrodeposition. Nanoporosity and nanostructure were observed as formation of nanocomposite. The nano-(HA and ACP) phases were essentially composed of a nanoporous and nanostructural biocomposite. Reconstituted collagen incorporation with the nanoporous and nanostructural biocomposite significantly facilitated the osteogenic differentiation of mesenchymal stem cells. Reconstituted collagen was covered with nano-(HA and ACP), profoundly impacting the enhancement of biocompatibility on application of implant and tissue engineering. The bioactive nano-HA/reconstituted collagen-induced osteogenic differentiation of mesenchymal stem cells enables to enhance bone growth/repair and osseointegration. PMID- 19283832 TI - Natural bone collagen scaffold combined with OP-1 for bone formation induction in vivo. AB - The scaffold is a key element to osteogenic tissue engineering as it provides a microenvironment for bone formation. Natural bone collagen scaffold (NBCS) is a novel biomaterial scaffold acid-extracted from organic human bone. The objective of this study was to characterize NBCS and evaluate the osteoconductivity of the scaffold, in combination with osteogenic protein-1 (OP-1), using a rabbit posteolateral lumbar fusion model. Thirty two rabbits were divided into 4 experimental groups, autograft, NBCS alone, OP-1 alone or NBCS combined with OP 1. Bone formation was evaluated by micro-CT, quantitative histological analysis, immunohistochemistry and semi-quantitative RT-PCR at 6 weeks postoperatively. By scanning electronic microscope, we showed that NBCS maintains a porous, interconnecting microarchitecture. Micro-CT analysis demonstrated that NBCS combined with OP-1 significantly induced (p < 0.01) bone formation at the fusion site as compared to control groups. This was confirmed by quantitative histological analysis which demonstrated that the NBCS combined with OP-1 significantly enhanced bone matrix area (17.7 mm(2)) (p < 0.05) and bone marrow cavity size (71.3 mm(2)) (p < 0.05) as compared to the controls. Immunohistochemical assessment and RT-PCR also demonstrated that NBCS combined with OP-1 enhanced type I collagen and osteonectin expression. Together, these results suggest that NBCS is an effective scaffold for osteogenesis, and combined with growth factors such as OP-1, possesses both osteoconductive and osteoinductive properties that are sufficient for bone regeneration. PMID- 19283833 TI - Biodegradable beta-Tri-Calciumphosphate/hydroxyethyl methacrylate enhanced three component bone adhesive demonstrates biocompatibility without evidence of systemic toxicity in a rabbit model. AB - Bone gluing is an attractive surgical technique; however, its use in patients is hampered by a variety of side effects. Therefore, it was the aim of this ethically approved study to evaluate a novel biodegradable beta-Tri Calciumphosphate (beta-TCP, Cerasorb)-enhanced bone adhesive regarding its toxicity and biocompatibility in a rabbit model. Fifty healthy New Zealand White rabbits were assigned in the study (n = 21) and sham-operated control group (n = 29). In the study group, a cylindrical part (4.6 x 10.0 mm) of the proximal tibia and distal femur was removed, reimplanted, and bone adhesive was applied. Blinded physical examination and sampling for hematology, clinical chemistry, and acute phase proteins (haptoglobin, C-reactive protein (CRP)) was performed before surgery and after 12, 24, 48, 72, 240, and 504 h. Significant findings of the physical examination included a slightly higher grading of warmth (p = 0.0019) and pain (p < 0.0001) of the wound 240 h after surgery in the study group. No significant differences between albumin, haptoglobin, CRP, and urea concentrations in both groups were evident, whereas the study group demonstrated significantly lower leukocyte count, total protein, and globulin concentrations (p < 0.0001). As expected, both groups showed a marked transient increase in muscle enzymes (Creatine Kinase and Aspartate Aminotransferase) following the surgery (p < 0.0001). Twelve to 24 h after surgery, a significant decrease in ionized calcium from 1.38 +/- 0.12 mmol/L to 1.06 +/- 0.13 mmol/L was noted in the study group (p < 0.0001). The results clearly indicated that the novel beta TCP-enhanced bone adhesive showed good biocompatibility without significant evidence of acute or subacute local or systemic toxicity. PMID- 19283834 TI - Finding social benefits after a collective trauma: perceiving societal changes and well-being following 9/11. AB - Individuals frequently perceive positive changes in themselves following adversity; after a collective trauma, they may perceive such benefits in others or in their society as well. We examined perceived benefits of the September 11, 2001 (9/11) terrorist attacks in a 3-year study of a national sample of adults (N = 1382). Many individuals (57.8%) perceived social benefits of 9/11, including increased prosocial behavior, religiousness, or political engagement. Individuals who found increased national religiosity as a benefit 2 months post-9/11 reported greater positive affect and life satisfaction and lower distress and posttraumatic stress up to 3 years post-9/11. Pre-9/11 religiousness and Republican political affiliation predicted perceiving religion-related social benefits post-9/11. Perceptions of social change are important but understudied responses to stressful events. PMID- 19283835 TI - Posttraumatic anger in crime victims: directed at the perpetrator and at the self. AB - This study investigated the targets of anger that are most strongly involved in posttraumatic anger. Using a sample of 218 crime victims, the authors assessed the levels of anger at potential targets (perpetrator, criminal justice system, third persons, and the self) and their association with severity of posttraumatic stress disorder (PTSD) symptoms. The results revealed that anger was most strongly directed at the perpetrator and at the self, and that anger at other targets was low. Moreover, anger at the perpetrator and at the self showed strong associations with PTSD symptoms, whereas the associations of anger at other targets with PTSD symptoms were low. PMID- 19283836 TI - Clinic and radiological improvement of lipoid pneumonia with multiple bronchoalveolar lavages. AB - OBJECTIVE: To assess the potential role of multiple bronchoalveolar lavages (BALs) in the treatment of children with lipoid pneumonia (LP). MATERIALS AND METHODS: This prospective study included 10 children (7 female, 3 male) with LP secondary to mineral oil aspiration. The age ranged from 3 months to 7 years and 1-60 days history of mineral oil intake, with a 6 months clinic follow-up. High resolution computer tomography (CT) was performed 1-7 days prior to treatment and 2-20 days after the last therapeutic BAL, and reviewed by two experienced chest radiologists. Oxygen saturation was measured with digital oximetry. Therapeutic BAL was performed weekly until BAL fluid was nearly transparent and the cell count returned to normal range values. RESULTS: In all children, the initial CT scans showed multifocal bilateral consolidation involving mainly the dorsal and central regions. The areas of consolidation had foci of decreased attenuation in eight patients. Following a total of 4-10 therapeutic BALs, the CT scans returned to normal in 3 patients, improved considerably in 5, and showed only slight improvement in 2. Oxygen saturation increased from 88.8 +/- 3.4% at presentation to 96.2 +/- 0.8% after treatment (P < 0.0001). Multiple lavages reduced (P < 0.003) numbers of lipid-laden macrophages and restored BAL cellularity to normal range values. CONCLUSION: Multiple therapeutic BAL of children with LP results in significant improvement of CT findings, oxygen saturation, restoration of BAL fluid cellularity and clinical recover without any evidence of respiratory distress at the end of treatment and 6 months after the last BAL. PMID- 19283837 TI - Paralysis and a perihilar protuberance: an unusual presentation of sarcoidosis in a child. AB - Sarcoidosis in children can present with extra-pulmonary manifestations, making diagnosis difficult. We describe a case of sarcoidosis in a child, presenting as a Guillain-Barre-like illness with the incidental finding of a perihilar mass. We also report the first successful use of the minimally invasive technique of Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration (EBUS-TBNA) to delineate the cause of hilar lymphadenopathy in a child. PMID- 19283838 TI - Predictors of major intervention in infants with bronchiolitis. AB - OBJECTIVE: We sought to identify predictors of the major medical intervention (MMI) in infants with bronchiolitis in the Emergency Department (ED) to recognize those in need of hospitalization versus the candidates for discharge. PATIENTS AND METHODS: We conducted an analysis of data from a prospective cohort study of previously healthy infants 2-23 months presenting to our ED with first episode of wheeze and respiratory distress. Infants were divided into those with at least one MMI defined as oxygen administration for saturation of <90%, intravenous (IV) fluids of 20 ml/kg, apnea management, or critical care unit (CCU) admission (MMI group) versus those without (no-MMI group). The primary outcome was the association between the MMI versus no-MMI groups and potential risk factors for these outcomes. RESULTS: Of 312 study infants, 52 experienced MMI--all received oxygen for saturation <90%, four also received IV fluids and none required apnea management or CCU care. The following four risk factors were associated with MMI: baseline accessory muscle score >or=6/9 [OR 2.44, 95% CI 1.29; 4.62], oxygen saturation or=60 [OR 1.85, 95% CI 0.97; 3.54], and poor fluid intake [OR 2.65, 95% CI 1.12; 6.26]. Of the 148 infants without predictors 11 (7.4%) received MMI, 145 required either no MMI or oxygen for G in exon 1 of the LMNA gene, resulting in a leucine to arginine change at codon 59 (Leu59Arg). Mutations in the LMNA gene cause a variety of disorders including dilated cardiomyopathy, muscular dystrophy, familial lipodystrophy, progeria, atypical progeroid syndromes, and mandibuloacral dysplasia. Genotype-phenotype correlation has been reported for some of these conditions. Our patients are the only ones known to have the specific mutation Leu59Arg and also share a set of features not entirely consistent with any of the laminopathies previously described. A previously reported patient with an adjacent mutation (Ala57Pro) had "atypical Werner syndrome" with dilated cardiomyopathy, hypogonadism, and sloping shoulders. While each of these clinical features does occur in other laminopathy syndromes, these patients form a phenotypic cluster distinct from other laminopathies and clinically overlapping with Malouf syndrome. LMNA sequencing should be considered for patients presenting with dilated cardiomyopathy and hypergonadotropic hypogonadism, including those previously diagnosed with Malouf syndrome. PMID- 19283855 TI - Allelic heterogeneity in inbred populations: the Saudi experience with Alstrom syndrome as an illustrative example. AB - The increased frequency of rare autosomal recessive conditions in genetically isolated populations is a well-established phenomenon. This genetic isolation is invoked as an explanation when one particular mutation is the sole or most frequent mutation observed in a given population and is referred to as the founder effect. This trend of allelic homogeneity is contrasted by an opposite trend when the consanguinity factor is in play. Independent of endogamy at the population level, a consanguineous union is sufficient to render homozygous a percentage of the genome that is directly correlated with the degree of consanguinity. Assuming the gene in question has a normal mutation rate, the resulting homozygosity will inevitably include different defective alleles of that gene. By reporting four novel alleles, we use Alstrom disease to exemplify the interesting observation of allelic heterogeneity for a very rare autosomal recessive disorder in a highly inbred population. While we frequently assume founder effect in inbred populations, this report should serve to remind us of the powerful effect of the consanguinity factor, a common confounding variable among some of those populations. PMID- 19283856 TI - Gardner-Silengo-Wachtel or genito-palato-cadiac syndrome with associated autosomal aneuploidy. AB - Gardner-Silengo-Wachtel or genito-palato-cardiac syndrome is a disorder of male (46,XY) gonadal dysgenesis, thought to be either an X-linked recessive or an autosomal recessive disorder. The propositus in our report presented with multiple congenital anomalies including micrognathia, cleft palate, congenital heart defect with D-transposition, double outlet right ventricle, PFO, VSD, PDA and pulmonary valve stenosis and gonadal dysgenesis. Chromosome analysis showed a 46, XY, t(1;7)(q32,q22.1) der(10) t(3;10) (q21;q26)pat karyotype. This represents a rare case of autosomal aneuploidy associated with Gardner-Silengo-Wachtel or genito-palato-cardiac syndrome and suggests genetic heterogeneity for this syndrome. Partial monosomy of 10q also shares many of the prominent features of genito-palato-cardiac syndrome, including gonadal dysgenesis, cardiac defects and facial features. Monosomy for distal 10q may present as a phenocopy of Gardner Silengo-Wachtel or genito-palato-cardiac syndrome. Alternatively, unmasking of a recessive allele on distal 10q may result in genito-palato-cardiac syndrome, thus potentially localizing a candidate region for the gene to 10q26 --> qter. PMID- 19283857 TI - Connexin mutations in Brazilian patients with skin disorders with or without hearing loss. AB - The connexins are a family of proteins whose major function is as part of the gap junctions of cell-to-cell channels. They are expressed in several tissues including brain, skin, and cochlea. Mutations in connexin genes play a major role in non-syndromic sensorineural deafness, but have been also described in individuals with variable dermatological features. In recent years, many genes responsible for hereditary skin diseases have been discovered. These genes may encode different proteins that participate in the terminal differentiation of the epidermis. Therefore alteration or absence of these proteins causes a keratinization disorder. It has been demonstrated that distinct germline mutations within six connexin (Cx) genes GJB2 (Cx26), GJB6 (Cx30), GJB3 (Cx31), GJA1 (Cx43), GJB4 (Cx30.3), and GJB5 (Cx31.1), may cause sensorineural hearing loss and various skin disease phenotypes. The crucial functional importance of each of these connexins in the mentioned ectodermic tissues is reflected by the finding that genetic defects in their genes produce a wide spectrum of genetic disorders comprising sensorineural hearing loss, disorders of cornification of the skin, hair, and nails, and keratitis. Here, we report on different mutations in the connexin genes in individuals with or without hearing loss and different skin disorders illustrating the clinical and genetic heterogeneity of the condition. PMID- 19283858 TI - Toxicology evaluation of Atlantic Canadian seafood processing plant effluent. AB - The purpose of this study was to carry out an acute aquatic toxicity assessment on select effluent samples from Atlantic Canadian seafood processing plants. Raw effluent acute aquatic toxicity for the flatfish and salmon effluents was assessed using the acute lethality test and Microtox test. The effectiveness of dissolved air flotation treatment (DAF) in removing acute toxicity from these effluents was evaluated using the Microtox test. The salmon effluent failed the acute lethality test using rainbow trout while the flatfish effluent showed acute toxicity in the Microtox test with a 50% inhibiting concentration (IC(50)) of 38.84%. Subsequent treatment by DAF of the flatfish and salmon effluents increased IC(50) values by 20% and 26% respectively. The findings of this study indicate that all of the processing effluents sampled showed characteristics that could potentially degrade effluent receiving waters and acute toxicity was demonstrated in the two raw finfish effluents. Application of DAF treatment was successful in significantly increasing Microtox IC(50) values, thereby reducing acute toxicity, but failed to entirely remove acute toxicity. PMID- 19283859 TI - Individual and family adaptation in Taiwanese families of individuals with severe and persistent mental illness (SPMI). AB - The purpose of this study was to examine individual and family adaptation in Taiwanese families of persons with severe and persistent mental illness. Mediating effects of social support and meaning of family caregiving on the relationship between pile-up of demands and family adaptation were also assessed. Data were collected from 84 families using mailed questionnaires and analyzed using principal component analysis and mixed linear modeling. Adaptation was associated with lower pile-up of demands, greater social support, and more positive interpretation of family caregiving. Partially mediating effects of social support and meaning of family caregiving were supported. The results imply a need for developing culturally sensitive interventions to decrease family demands and amplify individual, family, and community strengths and resources. PMID- 19283860 TI - Postnatal development in cynomolgus monkeys following prenatal exposure to natalizumab, an alpha4 integrin inhibitor. AB - BACKGROUND: Natalizumab is a humanized monoclonal IgG4 antibody to human alpha4 integrin that blocks the interaction of alpha4beta1 and alpha4beta7 integrins with their ligands, including fibronectin, vascular cell adhesion molecule-1, and mucosal addressin cellular adhesion molecule-1. Because alpha4 integrins and their ligands are widely involved in mammalian development, lymphopoeisis, and hematopoiesis, natalizumab may interfere with these processes. METHODS: The effects of prenatal exposure to natalizumab on postnatal development were assessed in cynomolgus monkeys at doses of 0 and 30 mg/kg administered intravenously every other day from gestational day (GD) 20 to 70 or GD 20 to term. Infants were delivered by natural birth and evaluated for general health, survival, development, and immunological structure and function at 12 or 18 months. RESULTS: An increase in abortions was seen in the first cohort of natalizumab-treated dams (39.3 vs. 7.1% in the controls) but not in the second cohort (33.3, 37.5%). Infants in the term treatment group had elevated lymphocyte ( approximately 150%) and nucleated red blood cell counts ( approximately 400%), consistent with the pharmacological effect of natalizumab, and reductions in platelet counts ( approximately 28%), which were reversible following clearance of natalizumab. No anemia was observed. Infants in the term treatment group had significantly increased spleen weights at 12 months but not at 18 months. All other experimental observations in infants from natalizumab-treated dams were comparable with those of controls. CONCLUSION: Natalizumab had no adverse effects on the general health, survival, development, or immunological structure and function of infants born to dams treated with natalizumab during pregnancy. PMID- 19283861 TI - The effects of Scutellaria baicalensis extract on embryonic development in mice. AB - BACKGROUND: Scutellaria baicalensis is a perennial herbaceous plant widely distributed in Oriental areas. Its roots, a commonly used medicinal source, reputedly calm fetuses in pregnant women; however, there is no sufficient evidence to date to assess its safety during pregnancy. This study aims to evaluate the effects of S. baicalensis aqueous extract on embryonic development in ICR mice. METHODS: Aqueous extract of S. baicalensis roots was prepared in accordance with clinical application. Pregnant mice were randomly divided into four groups, i.e., mice treated by gavage with water as negative control, with aqueous extract of 2 (1.8 times of human daily dose), 8 or 32 g/kg/day from gestation day (Gd) 6 to 15 as low-, middle-, and high-dose groups, respectively. The parameters of live and dead fetuses, resorptions, external and skeletal malformed fetuses, maternal body weight, maternal liver, kidneys, and heart weights were evaluated on Gd 18. RESULTS: There was no significant difference in fetal parameters among four groups. Maternal absolute liver and kidneys weights in the high-dose group were significantly higher than those in negative control (p<0.05). Relative liver and kidneys weights in this group were significantly higher than those in any other group (p<0.05). CONCLUSIONS: Oral administration of aqueous extract of S. baicalensis roots at or below 32 g/kg/day to ICR mice during organogenesis did not cause significant fetal external or skeletal malformations. However, 32 g/kg/day presented potential maternal toxicity. PMID- 19283862 TI - Effects of natalizumab, an alpha4 integrin inhibitor, on fertility in male and female guinea pigs. AB - BACKGROUND: Natalizumab is a humanized monoclonal immunoglobulin G4 antibody directed against the human alpha4 integrin subunit disrupting interaction with its ligands. As alpha4 integrins and/or their ligands appear to be involved in reproductive function, the effects of natalizumab on fertility in male and female guinea pigs were investigated. METHODS: Natalizumab was administered by bolus intravenous injection every other day at doses of 0, 3, 10, and 30 mg/kg. Males began treatment at least 28 days prior to mating until necropsy (approximately 3 to 5 days after mating). Dosing in females was done from gestational day (GD) of an existing pregnancy to GD 30 of a second pregnancy. RESULTS: In male guinea pigs, natalizumab treatment had no effect on sperm parameters, reproductive organ weights, organ-weight ratios, or histology of the testis or epididymis. Natalizumab did not affect the ability of treated males to produce pregnancies in untreated females. In female guinea pigs, no treatment-related changes were seen in uterine weights or ovary weights. Pregnancy rates were reduced in females treated with 30 mg/kg natalizumab, but not those treated with 3 or 10 mg/kg. Pregnancy rates were 63.3, 66.7, 66.7, and 29.6% for groups treated with 0, 3, 10, and 30 mg/kg, respectively. Effects observed at 30 mg/kg were at exposures 36 fold those observed in humans. CONCLUSIONS: Natalizumab had no effects on male fertility, but did result in a reduction in pregnancy rates in females treated with the high dose of 30 mg/kg. PMID- 19283863 TI - Urodynamic findings in female diabetic patients with and without overactive bladder symptoms. AB - AIMS: The purpose was to analyze urodynamic findings in female diabetic patients with OAB symptoms. METHODS: Data from 94 female diabetic patients who underwent urodynamic studies in evaluation of various LUTS were retrospectively reviewed. Urodynamic findings, demographic data, and clinical symptoms were compared between patients with and without OAB. RESULTS: Among the 94 subjects analyzed, 34 (36.2%) were diagnosed as OAB. Demographic data were similar between the patients with and without OAB. In the OAB group, patients had significantly higher storage symptom scores and marginally higher voiding symptom scores. On cystometry, the OAB group had a higher percentage of increased bladder sensation (41.2% vs 11.7%, P = 0.001) and detrusor overactivity (29.4% vs 10.0%, P = 0.023). The OAB group had lower peak flow rate (16.2 +/- 5.9 vs 19.3 +/- 6.3 ml/s, P = 0.023), greater PVR volume (60.3 +/- 29.4 vs 45.0 +/- 25.1 ml, P = 0.009), and lower bladder voiding efficiency (BVE, 75.2 +/- 2.8 vs 81.5 +/- 2.9%, P < 0.001). On pressure-flow studies, the OAB group had a higher percentage of BOO (26.5% vs 6.7%, P = 0.008). CONCLUSIONS: Our study shows that the most frequent urodynamic finding of OAB in female diabetic patients is increased bladder sensation, followed by detrusor overactivity. Compared to those without OAB, female diabetic patients with OAB are more likely to have impaired voiding function, characterized by lower peak flow rate, greater PVR volume, lower BVE, and a higher percentage of BOO. In these patients, BOO not only causes voiding difficulty but may also contribute to the development of OAB. PMID- 19283864 TI - Bladder urothelial cells from patients with interstitial cystitis have an increased sensitivity to carbachol. AB - AIMS: The presence of muscarinic receptors on bladder urothelial cells (BUC), suggests BUC may be a target for antimuscarinics. This study determined whether human BUC are responsive to a muscarinic agonist and if so, whether responses are altered in interstitial cystitis (IC) BUC. METHODS: Primary urothelial cell cultures were established from cystoscopic biopsies. Normal (NB) and IC BUC were studied using calcium imaging techniques as a means to monitor the response to muscarinic receptor activation with the agonist, carbachol (CCh). Changes in intracellular Ca(2+) concentration ([Ca(2+)](i)) were measured with fura-2 ratiometric microfluorimetry. Dose-response curves (CCh dose vs. [Ca(2+)](i)) were measured in IC and NB BUC. Tolterodine was used to confirm the specificity (muscarinic versus nicotinic) of CCh evoked increases in [Ca(2+)](i). RESULTS: CCh induced a dose-dependent increase in [Ca(2+)](i). Potency and efficacy of CCh was significantly greater in IC BUC. The maximal increase in [Ca(2+)](i) was 136.3 +/- 5.1% over baseline in 78 cells from 4 IC patients versus 92.4 +/- 4.8% over baseline in 67 cells from 4 NB subjects (P < 0.01). The EC50 of the evoked increase was 1.10 +/- 0.14 microM versus 3.36 +/- 0.72 microM (P < 0.01) in BUC from IC and NB controls, respectively. Removal of extracellular calcium or application of tolterodine, abolished CCh evoked increase in [Ca(2+)](i) in IC and NB BUC. CONCLUSIONS: The greater sensitivity of IC BUC to CCh suggests that IC patholobiology may also include alterations muscarinic signaling. The physiologic sequelae of muscarinic activation in BUC need to be further investigated. PMID- 19283865 TI - An extract (THC-002) of Ba-Wei-Die-Huang-Wan inhibits expression of tachykinins, and P2X3 and TRPV1 receptors, and inhibits ATP-induced detrusor overactivity in spontaneously hypertensive rats. AB - AIMS: To investigate possible mechanisms of action of THC-002 (HARNCARE), a galenical produced from the traditional Chinese herbal mixture Ba-Wei-Die-Huang Wan, which has been reported to improve lower urinary tract symptoms (LUTS) in patients. METHODS: Forty-five female SHRs were randomly separated into three groups. Two groups were given 20 ml physiological saline solution (PSS) per kg body weight orally daily for 1 week. An hour after the administration of PSS, one of the groups received 20 mg THC-002 per kg body weight, and the other a similar volume of THC-002-free saline. The third group received no treatments. The bladders were analyzed by real time RT-PCR (n = 6) and immunohistochemistry (n = 3) for the expression of tachykinins and P2X3 and TRPV1 receptors. Cystometric investigation (n = 6) was conducted after intravesical instillation of saline followed by 5 mg/ml ATP solution. RESULTS: Treatment with PSS caused and upregulation of tachykinins and P2X3 and TRPV1 receptors, which was prevented in the group treated with THC-002. In the normal (non-treated) and non-THC-002 treated SHRs, instillation of the ATP solution decreased voiding interval, micturition volume, and bladder capacity compared to the instillation of saline. However, in the THC-002-treated SHRs, ATP instillation had no effect. CONCLUSIONS: In SHRs, THC-002 reduced the bladder expression of tachykinins and P2X3 and TRPV1 receptors, and inhibited ATP-induced detrusor overactivity. These effects may explain part of its beneficial effects on LUTS. PMID- 19283866 TI - Pelvic floor stimulation: what are the good vibrations? AB - OBJECTIVE: The aim of this study was to determine if two different whole body vibration, sinusoidal vibration (SV) and stochastic resonance vibration (SRV), using various intensities lead to a reactive activation of pelvic floor muscles. STUDY DESIGN: We compared the pelvic floor muscle response of a healthy control group with that of a post partum group with weakened pelvic floor contraction. Activation effects of stochastic resonance vibration and sinusoidal vibration with six increasing vibration intensities were investigated using pelvic floor EMG and compared to activity during rest and maximum voluntary contraction. RESULTS: Both whole body vibration systems were able to activate pelvic floor muscles significantly depending on vibration intensity. Generally, the SRV achieved a significantly higher activation than maximum voluntary contraction, especially in women post partum and using a frequency of 6-12 Hz. CONCLUSION: SRV, compared to SV, leads to higher pelvic floor muscle activation in subjects with weakened pelvic floor muscles and achieves higher pelvic floor activation than maximum voluntary contraction alone. Neurourol. Urodynam. 28:405-410, 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19283867 TI - Suppression of reflex urethral responses by sacral dermatome stimulation in an acute spinalized feline model. AB - AIMS: Reflex contractions of the external urethral sphincter (EUS) are a major component of voiding dysfunction after neurological injury or disease. Aberrant urethral reflexes can prevent voiding and cause serious medical complications. Characterizing these urethral reflexes during genitourinary studies is necessary for evaluating novel pharmacological or neuroprosthetic approaches. The objectives of the present study were to generate urethral reflexes in the acute spinal feline, to quantify these reflexes, and to suppress them with electrical stimulation of the sacral dermatomes. METHODS: This study comprised eight male cats. Anaesthesia was maintained with alpha-chloralose or sodium pentobarbital. The spinal cord was transected between T10 and T12, and nerve cuff electrodes were placed on the extradural S2 sacral roots to provide bladder activation. Bladder and urethral pressures were recorded during and after bladder contractions. Electrical stimulation was applied non-invasively to the sacral dermatomes with commercial surface electrodes. RESULTS: Urethral reflexes were elicited consistently in six cats. The corresponding urethral pressure spikes were quantified. Putative metrics of urethral reflex activity such as the rate and average magnitude of reflex pressure spikes correlated significantly with standard urodynamic variables. Electrical stimulation of the sacral dermatomes suppressed urethral reflexes in three cats. CONCLUSIONS: These findings in an acute spinal feline preparation demonstrate a non-invasive means of suppressing undesirable urethral reflexes. Translation of this work to clinical use could improve neuroprostheses for restoring bladder function and enhance treatment of aberrant urethral reflexes in humans. PMID- 19283868 TI - Patient reported outcomes tools in an observational study of female stress urinary incontinence. AB - AIMS: To determine which patient characteristics, incontinence and non incontinence related, are associated with the symptom severity scores of the Urogenital Distress Inventory (UDI) and the International Consultation on Incontinence Questionnaire Urinary Incontinence (ICIQ-UI); and to determine the association of both patient characteristics and symptom severity scores with quality-of-life scores of the Incontinence Impact Questionnaire (IIQ) and the Incontinence-Quality of Life (I-QOL) questionnaire. METHODS: Women presenting with stress urinary incontinence (SUI) symptoms in primary and secondary care entered the Stress Urinary Incontinence Treatment Study (SUIT), an observational study evaluating the cost-effectiveness of duloxetine compared to other non surgical treatments for SUI. At enrollment patients completed the UDI-6, the short form ICIQ-UI, the IIQ-7 and the I-QOL. Multivariate linear regressions were performed with the UDI-6, ICIQ-UI SF, IIQ-7, and I-QOL as outcomes. RESULTS: The total number of incontinence episodes is the most significant explanatory variable of the two symptom questionnaire scores, but the UDI-6 score also reflects the type of incontinence. The variability of the condition-specific quality-of-life questionnaires is primarily explained by the symptom severity questionnaire scores. Although there is a high intercorrelation, both these symptom questionnaires independently contributed significantly to the IIQ-7 and I QOL total scores. CONCLUSIONS: The UDI-6 and ICIQ-UI SF can be regarded as scientifically sound symptom questionnaires in UI evaluation; but they have differences. Since the UDI-6 and ICIQ-UI SF independently contribute to the quality-of-life scores, this suggests that in incontinence research symptom questionnaires should not focus only on incontinence, but on a broader range of urogenital symptoms. PMID- 19283869 TI - Anti-angiogenesis effect of generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide on breast cancer in vitro. AB - OBJECTIVE: To study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide (G4PAMAM/VEGFASODN) compound on the expressions of vascular endothelial growth factor (VEGF) and its mRNA of breast cancer cells and on the inhibition of vascular endothelial cells. METHODS: We examined the morphology of G4PAMAM/VEGFASODN compound and its pH stability, in vitro transfection efficiency and toxicity, and the expressions of VEGF and its mRNA. Methyl thiazolyl tetrazolium assay was used to detect the inhibitory function of the compound on vascular endothelial cells. RESULTS: The compound was about 10 nm in diameter and was homogeneously netlike. From pH 5 to 10, it showed quite a buffered ability. The 48-h transfection rate in the charge ratio of 1:40 was 98.76%, significantly higher than that of the liposome group (P<0.05). None of the transfection products showed obvious toxicity on the cells. The expressions of both VEGF protein and its mRNA after G4PAMAM/VEGFASODN transfection decreased markedly. CONCLUSION: With a low toxicity, high safety, and high transfection rate, G4PAMAM/VEGFASODN could be a promising gene vector. Specifically, it inhibits VEGF gene expression efficiently, laying a basis for further in vivo animal studies. PMID- 19283870 TI - Is adult celiac disease really uncommon in Chinese? AB - Celiac disease (CD) is a type of intestinal malabsorption syndrome, in which the patients are intolerant to the gliadin in dietary gluten, resulting in chronic diarrhea and secondary malnutrition. The disease is common in Europe and the United States, but only sporadic reports are found in East Asia including China. Is CD really rare in China? We examined 62 patients by capsule endoscopy for chronic diarrhea from June 2003 to March 2008. Four patients with chronic diarrhea and weight loss were diagnosed to have CD. Under the capsule endoscopy, we observed that the villi of the proximal small bowel became short, and that the mucous membrane became atrophied in these four patients. Duodenal biopsies were performed during gastroscopy and the pathological changes of mucosa were confirmed to be Marsh 3 stage of CD. A gluten free diet significantly improved the conditions of the four patients. We suspect that in China, especially in the northern area where wheat is the main food, CD might not be uncommon, and its under-diagnosis could be caused by its clinical manifestations that could be easily covered by the symptoms from other clinical situations, particularly when it came to subclinical patients without obvious symptom or to patients with extraintestinal symptoms as the initial manifestations. PMID- 19283871 TI - Enhancive effect of N,N'-dinitrosopiperazine on inducing precancerous lesion on nasal and/or nasopharyngeal epithelia of TgN(p53mt-LMP1)/HT mice. AB - OBJECTIVE: To investigate the enhancive effect of N,N'-dinitrosopiperazine (DNP) on induced carcinogenesis in nasal and/or nasopharyngeal epithelia among TgN(p53mt-LMP1)/HT transgenic mice to examine the underlying mechanism for the development of nasopharyngeal carcinoma (NPC). METHODS: TgN(p53mt-LMP1)/HT transgenic mice and the same strain of C(57)BL/6J wild-type mice both at the age of 5 months were randomly divided into 2 groups in parallel, respectively, i.e., TgN(p53mt-LMP1)/HT cancerous lesion-inducing group (TI), TgN(p53mt-LMP1)/HT control group (TC), C57BL/6J cancerous lesion-inducing group (CI), and C57BL/6J control group (CC). TI and CI mice were treated only with DNP for 16 weeks, twice each week, while TC and CC mice were given the same volume of saline as controls. At the end of treatment, animals were sacrificed to collect epithelial tissue samples from nasal cavity and nasopharynx for pathohistological evaluation by haematoxylin and eosin (HE) staining and for determination on the expression of TRAF2, c-Jun, and p16 by immunohistochemistry. RESULTS: Atypical hyperplasia was more significant in the samples of TI than in those of TC, CI, and CC, with the rates of lesions being 90%, 10%, 0, and 0 (P<0.01) respectively, though DNP was used alone in a much shortened inducing period at less dosage and without the use of carcinogenic promoter 12-O-tetradecanoylphorbol-13-acetate as usual. The expressions of tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2) and c-Jun in these samples were significantly up-regulated in TI (P<0.01), while the expression of p16 was significantly lower in TI than in the other groups (P<0.01). CONCLUSION: TgN(p53mt-LMP1)/HT mice hold inherited constitutional defect in immune surveillance function, which can be aggravated by environmental carcinogens, such as DNP used even though in a much less strength. The enhanced carcinogenesis-inducing effect of DNP on TgN(p53mt-LMP1)/HT mice should be closely associated with abnormal signaling of activator protein-1 (AP-1) pathway, especially up-regulated expressions of TRAF2 and c-Jun, and down-regulated expression of p16. PMID- 19283872 TI - Spinal cord decompression reduces rat neural cell apoptosis secondary to spinal cord injury. AB - OBJECTIVE: To determine whether spinal cord decompression plays a role in neural cell apoptosis after spinal cord injury. STUDY DESIGN: We used an animal model of compressive spinal cord injury with incomplete paraparesis to evaluate neural cell apoptosis after decompression. Apoptosis and cellular damage were assessed by staining with terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) and immunostaining for caspase-3, Bcl-2 and Bax. METHODS: Experiments were conducted in male Sprague Dawley rats (n=78) weighing 300-400 g. The spinal cord was compressed posteriorly at T10 level using a custom-made screw for 6 h, 24 h or continuously, followed by decompression by removal of the screw. The rats were sacrificed on Day 1 or 3 or in Week 1 or 4 post-decompression. The spinal cord was removed en bloc and examined at lesion site, rostral site and caudal site (7.5 mm away from the lesion). RESULTS: The numbers of TUNEL-positive cells were significantly lower at the site of decompression on Day 1, and also at the rostral and caudal sites between Day 3 and Week 4 post-decompression, compared with the persistently compressed group. The numbers of cells between Day 1 and Week 4 were immunoreactive to caspase-3 and B-cell lymphoma-2 (Bcl-2)-associated X-protein (Bax), but not to Bcl-2, correlated with those of TUNEL-positive cells. CONCLUSION: Our results suggest that decompression reduces neural cell apoptosis following spinal cord injury. PMID- 19283873 TI - Effects of intermittent negative pressure on osteogenesis in human bone marrow derived stroma cells. AB - OBJECTIVE: We investigated the effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells (BMSCs) in vitro. METHODS: BMSCs were isolated from adult marrow donated by a hip osteoarthritis patient with prosthetic replacement and cultured in vitro. The third passage cells were divided into negative pressure treatment group and control group. The treatment group was induced by negative pressure intermittently (pressure: 50 kPa, 30 min/times, and twice daily). The control was cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast microscopy, the determination of alkaline phosphatase (ALP) activities, and the immunohistochemistry of collagen type I. The mRNA expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) in BMSCs were analyzed by real-time polymerase chain reaction (PCR). RESULTS: BMSCs showed a typical appearance of osteoblast after 2 weeks of induction by intermittent negative pressure, the activity of ALP increased significantly, and the expression of collagen type I was positive. In the treatment group, the mRNA expression of OPG increased significantly (P<0.05) and the mRNA expression of OPGL decreased significantly (P<0.05) after 2 weeks, compared with the control. CONCLUSION: Intermittent negative pressure could promote osteogenesis in human BMSCs in vitro. PMID- 19283874 TI - Protective effects of Salvia miltiorrhizae on the hearts of rats with severe acute pancreatits or obstructive jaundice. AB - OBJECTIVE: To investigate the therapeutic effects and mechanisms of Salvia miltiorrhizae (Danshen) in the treatment of severe acute pancreatitis (SAP)- or obstructive jaundice (OJ)-induced heart injury. METHODS: A total of 288 rats were used for SAP- (n=108) and OJ-associated (n=180) experiments. The rats were randomly divided into sham-operated, model control, and Salvia miltiorrhizae treated groups. According to the difference of time points after operation, SAP rats in each group were subdivided into 3, 6 and 12 h subgroups (n=12), whereas OJ rats were subdivided into 7, 14, 21, and 28 d subgroups (n=15). At the corresponding time points after operation, the mortality rates of the rats, the contents of endotoxin and phospholipase A2 (PLA2) in blood, and pathological changes of the hearts were investigated. RESULTS: The numbers of dead SAP and OJ rats in the treated groups declined as compared with those in the model control group, but not significantly (P>0.05). The contents of endotoxin (at 6 and 12 h in SAP rats and on 7, 14, 21, and 28 d in OJ rats, respectively) and PLA2 (at 6 and 12 h in SAP rats and on 28 d in OJ rats, respectively) in the treated group were significantly lower than those in the model control group (P<0.01 and P<0.001, respectively). Besides, myocardial pathological injuries were mitigated in SAP and OJ rats. CONCLUSION: In this study, we found that Salvia miltiorrhizae improved myocardial pathological changes, reduced the content of PLA2 in blood, and decreased the mortality rates of SAP and OJ rats, exerting protective effects on the hearts of the rats. PMID- 19283875 TI - Expressions of cardiac sympathetic norepinephrine transporter and beta1 adrenergic receptor decreased in aged rats. AB - Evidence suggests that the deterioration of communication between the sympathetic nervous system and cardiovascular system always accompanies the aging of human and animals. Cardiac sympathetic norepinephrine (NE) transporter (NET) on presynaptic membrane is a predominant component to eliminate released NE in the synaptic cleft and maintains the sensitivity of the beta-adrenergic receptor (beta-AR). In the present study, we investigated NET and beta1-AR mRNA levels and sympathetic nerve density in cardiac sympathetic ganglion and left ventricular myocardium in 2- and 16-month-old rats with Northern blot analysis and immunohistochemistry. The expression levels of NET mRNA, NET protein and beta1-AR mRNA in the ganglia or myocardia of 16-month-old rats were markedly reduced by 67%, 26%, and 43%, respectively, in comparison with those in 2-month-old rats. Our results also show that aging induces a strong decrease of the catecholaminergic nerve fiber density. PMID- 19283876 TI - Penn State Worry Questionnaire: structure and psychometric properties of the Chinese version. AB - The Penn State Worry Questionnaire (PSWQ) is a measure of worry phenomena and has been demonstrated valid in cross-cultural populations. The present study examined the factor structure and psychometric properties of the Chinese version of PSWQ (Ch-PSWQ) in a Chinese college sample (n=1243). Exploratory factor analysis of the Ch-PSWQ revealed a two-factor solution (engagement of worry and absence of worry). Confirmatory factor analysis and model comparison supported that the model of one factor with method effect provided the best fit to the data. The Ch PSWQ and its factors evidenced good internal consistency and both convergent and discriminate validity. The present study supports the opinion that the second factor of PSWQ not only contains the component of evaluating pathological worry, but also might represent other traits. PMID- 19283877 TI - Colorectal cancer, one entity or three. AB - Understanding of the mechanism of colorectal carcinogenesis has been gaining momentum for some years on account of its high incidence and impact on the lives of individuals affected. Different genetic abnormalities have been found in colorectal cancers from different sites. For example, proximal colon cancer is usually related to the nucleotide instability pathway, as microsatellite instability (MSI). However, distal colon cancer is usually associated with specific chromosomal instability (CIN). The development of cancer at the rectum, though similar to that at the colon, displays its own unique features. These differences might be partially attributed to different embryological development and physiological circumstances. Environmental factors such as diet and alcohol intake also differ in their role in the development of tumors in the three segments, proximal colon, distal colon, and rectum. "Proximal shift" of colon cancer has been known for some time, and survival rates of colorectal cancer are higher when rectal cancers are excluded, both of which emphasize the three different segments of colorectal cancer and their different properties. Meanwhile, colonic and rectal cancers are distinctive therapeutic entities. The concept of three entities of colorectal cancer may be important in designing clinical trails or therapeutic strategies. However, the dispute about the inconsistency of data concerning the site-specific mechanism of colorectal carcinoma does exist, and more evidence about molecular events of carcinogenesis and targeted therapy needs to be collected to definitely confirm the conception. PMID- 19283878 TI - Aorta-to-right atrium fistula, an unusual complication of endocarditis. AB - Infective endocarditis (IE) remains a serious disease. Aorta-to-right atrium fistula is a rare but very serious complication of IE and predicts a higher mortality. This report describes a 50-year-old man with endocarditis, vegetation, perforation of noncoronary sinus, and formation of two aorta-to-right atrium fistulas with native valves detected by transthoracic echocardiography. This disease is lethal despite developments in cardiac imaging and antibacterial therapy. Early diagnosis, aggressive antibacterial therapy, and surgical treatment may improve the prognosis. PMID- 19283879 TI - Chest wall abscess due to Prevotella bivia. AB - Prevotella bivia is associated with pelvic inflammatory disease. A 77-year-old man developed a rapidly growing chest wall abscess due to P. bivia within days. He underwent surgical resection of the infected area; his postoperative course was uneventful. This is the first case of chest wall abscess due to P. bivia infection. Its correct diagnosis cannot be underestimated because fulminant infections can occur in aged or immunocompromised patients if treated incorrectly. Prompt, appropriate surgical management, and antibiotic therapy affect treatment outcome. PMID- 19283880 TI - Gastric carcinoma with osteoclast-like giant cells: a case report and review of the literature. AB - Gastric carcinoma with osteoclast-like giant cells (OGCs) is an extremely rare tumor. So far, only six cases have been reported in the literature. Here we report an additional case of this tumor in a Chinese 78-year-old man presented with abdominal pain, vomiting, and hematemesis. Physical examination and gastroscopy revealed a tumor in the gastric antrum. The biopsy and pathological findings indicated a gastric adenocarcinoma with OGCs, which were present in both the tumor and the metastatic lymph nodes. Further immunohistochemical staining indicated that OGCs were reactive with CD68, CD45, and vimentin protein, but not with pancytokeratin, carcinoembryonic antigen, or epithelial membrane antigen, suggesting the monocytic/histiocytic derivation of these OGCs. In situ hybridization for Epstein-Barr virus showed no nuclear positivity in either adenocarcinoma or OGCs. Postoperative follow-up showed that the patient had survived for at least 6 months without recurrence. Further investigation is warranted to clearly define the prognostic significance of OGCs in gastric carcinoma. PMID- 19283881 TI - Growing role of CD40 ligand gene transfer therapy in the management of systemic malignancies besides hepatocellular carcinomas. PMID- 19283882 TI - Prosthetic valve endocarditis due to Brucella: successful outcome with a combined strategy. AB - Infective endocarditis is a serious heart disease that can affect native and prosthetic valves. Staphylococci and Streptococci are the etiological agents in almost 80% of cases. Human brucellosis is a multiple organ disease often transmitted via contaminated, unpasteurized goat milk and cheese. Brucella endocarditis is a rare complication associated with elevated morbidity and mortality rates; it can affect native or, even more rarely, prosthetic valves. In this report, we present the successful management of an infected aortic biological prosthesis in a man of 68 years. PMID- 19283883 TI - Postendocarditis mitral valve aneurysm. AB - Mitral valve aneurysm is defined as a localized, thin-walled, saccular bulge of the mitral leaflet toward the left atrium. In this study, we report a case of mitral aneurysm of the posterior leaflet (scallop P3) secondary to primary mitral endocarditis in a patient who underwent prosthetic aortic valve replacement and coronary artery bypass graft. PMID- 19283884 TI - A new approach to the assessment of marginal vitamin A deficiency in children in suburban Guwahati, India: hydrolysis of retinoyl glucuronide to retinoic acid. AB - The objective of the present study was to determine marginal vitamin A deficiency (VAD) by testing the hydrolysis of retinoyl glucuronide (RAG) to retinoic acid (RA) in children. Previous studies in rats showed that hydrolysis occurred when rats were vitamin A deficient. Children (n 61) aged 3-18 years, were divided into two groups, I and II. Blood was collected from the children in Group I (n 19) who were not dosed with RAG. Children in Group II (n 42) were administered all-trans retinoyl glucuronide (RAG) orally, and blood was collected 4 h after the dose. All serum samples were analysed for retinoids and carotenoids. RA was detected in serum only when serum retinol was < 0.85 micromol/l. Thus, hydrolysis of RAG to RA occurred in children with VAD or marginal VAD. Serum retinol was < 0.35 micromol/l in twenty-one children, 0.35-0.7 micromol/l in twenty-three children, 0.7-0.9 micromol/l in eleven children and >1 micromol/l in six children. Mean serum retinol in sixty-one children was 0.522 (sd 0.315) micromol/l. Mean beta carotene (0.016 (sd 0.015) micromol/l) was far below normal compared to the level of lutein (0.176 (sd 0.10) micromol/l) in sixty-one children. A low beta-carotene level might be due to a low intake of carotene but high demand for vitamin A. The RAG hydrolysis test may prove to be a useful approach for the determination of marginal VAD with no clinical or subclinical signs of VAD. High prevalence of VAD amongst certain communities in Assam cannot be ruled out. PMID- 19283885 TI - Ventricular fibrillation in acute mitral valve insufficiency caused by chordae tendineae rupture: report of a surgically corrected case. AB - In this report, we present the case of a previously healthy 80-year-old woman who was referred to surgery after a cardiac arrest due to ventricular fibrillation successfully resuscitated; the following evaluation revealed acute mitral regurgitation due to chordae tendineae rupture. After mitral valve repair, a single-chamber cardioverter-defibrillator was implanted for secondary prevention of sudden cardiac death. After 16 months of follow-up, the patient is asymptomatic without any further episodes of ventricular arrhythmias reported, underlying the pivotal role of mitral valve repair in the prevention of potentially lethal ventricular arrhythmias. PMID- 19283886 TI - Increased dietary protein consumed at breakfast leads to an initial and sustained feeling of fullness during energy restriction compared to other meal times. AB - The objective of the study was to assess whether the timing of increased dietary protein throughout the day influences the feelings of fullness during energy balance (EB) and restriction (ER). Nine men (age 48 (sem 6) years; BMI 32.7 (sem 0.7) kg/m2) randomly completed five controlled feeding trials, each consisting of 3 d of EB, followed by 3 d of ER of a 3138 kJ/d (750 kcal/d) reduction). The diet was composed of a normal amount of protein (NP) (0.8 g protein/kg per d), or an additional amount of protein (HP) (+0.6 g protein/kg per d) given at breakfast (HP-B), lunch (HP-L), dinner (HP-D) or equally divided among all meals (HP-E). Meal-related (3 h postprandial) and overall (15 h composite) feelings of fullness were assessed from thirteen-point, numbered, linear category scale questionnaires (reported as arbitrary units (au)). When comparing HP treatments, the data are presented as difference from NP. No differences in meal-related or overall fullness were observed among HP treatments during EB. During ER, the HP-B led to greater meal-related fullness (+137 (sem 44) au x 180 min) compared to HP-D ( -1 (sem 37) au x 180 min; P = 0.003), but not for HP-L (+62 (sem 53) au x 180 min; P = 0.188) or HP-E-B (+92 (sem 85) au x 180 min; P = 0.587). HP-B also led to greater overall (15 h) fullness (+404 (sem 162) au x 900 min) v. HP-L (+33 (sem 162) au x 900 min; P = 0.009) and HP-D ( -60 (sem 132) au x 900 min; P = 0.05), but not HP-E (+274 (sem 165) au x 900 min; P = 0.188). The initial and sustained feelings of fullness following protein consumption at breakfast suggests that the timing of protein intake differentially influences satiety during ER. PMID- 19283887 TI - In vivo test of the vertical phase separation hypothesis: the display of major histocompatibility complex (MHC) class I molecules on membranes of B cells from mice fed high-fat diets. AB - The membrane vertical phase separation hypothesis predicts that a decrease in plasma membrane acyl chain order will increase major histocompatibility complex (MHC) class I surface expression. The hypothesis is based on modification of plasma membrane acyl chain order in cell culture and has not been tested in vivo. In the present study, we isolated splenic B cells from C57/BL6 mice fed either a normal diet or high-fat diets enriched in SFA or MUFA and assayed for changes in plasma membrane acyl chain order and MHC class I surface expression. Plasma membranes of B cells from MUFA-fed mice had significantly decreased acyl chain order and increased headgroup order. The decrease in acyl chain order correlated with a significant increase in the acyl chain unsaturation of B cells from the MUFA-fed mice. MHC class I surface levels on B cells were not affected by the MUFA-rich diet. This study suggests that the membrane vertical phase separation hypothesis may have limited application in a physiologically relevant setting. PMID- 19283888 TI - Benazepril plus amlodipine or hydrochlorothiazide for hypertension. PMID- 19283889 TI - Benazepril plus amlodipine or hydrochlorothiazide for hypertension. PMID- 19283890 TI - Prolonged therapy for hepatitis C with low-dose peginterferon. PMID- 19283891 TI - Prolonged therapy for hepatitis C with low-dose peginterferon. PMID- 19283892 TI - Case 36-2008: a man with chronic daily headache. PMID- 19283893 TI - Mechanisms underlying the nociceptive responses induced by platelet-activating factor (PAF) in the rat paw. AB - Platelet-activating factor (PAF) is an inflammatory mediator widely known to exert relevant pathophysiological functions. However, the relevance of PAF in nociception has received much less attention. Herein, we have investigated the mechanisms underlying PAF-induced spontaneous nociception and mechanical hypersensitivity in the rat paw. PAF injection (1- 30 nmol/paw) resulted in a dose-related overt nociception, whilst only the dose of 10 nmol/ paw produced a significant and time-related mechanical hypersensitivity. Local coinjection of PAF antagonist WEB2086 significantly inhibited both spontaneous nociception and mechanical hypersensitivity. Moreover, the coinjection of the natural IL-1beta receptor antagonist (IRA) notably prevented both PAF-induced nociceptive responses, whilst these responses were not altered by anti-TNFalpha coinjection. Interestingly, pretreatment with the ultrapotent vaniloid agonist resiniferotoxin, coinjection of the TRPV1 receptor antagonist SB366791, or mast cell depletion with compound 48/80 markedly prevented PAF-induced spontaneous nociception. Conversely, PAF-elicited mechanical hypersensitivity was strikingly susceptible to distinct antineutrophil-related strategies, namely the antineutrophil antibody, the selectin blocker fucoidin, the chemokine CXCR2 receptor antagonist SB225002, and the C5a receptor antibody anti-CD88. Notably, the same antineutrophil migration strategies significantly prevented the increase of myeloperoxidase activity induced by PAF. The mechanical hypersensitivity caused by PAF was also prevented by the cyclooxygenase inhibitors indomethacin or celecoxib, and by the selective beta(1) adrenergic receptor antagonist atenolol. Collectively, the present results provide consistent evidence indicating that distinct mechanisms are involved in the spontaneous nociception and mechanical hypersensitivity caused by PAF. They also support the concept that selective PAF receptor antagonists might constitute interesting targets for the development of new analgesic drugs. PMID- 19283894 TI - Suramin inhibits the CD40-CD154 costimulatory interaction: a possible mechanism for immunosuppressive effects. AB - Suramin is a symmetric polysulfonated naphthylamine-benzamide urea derivative approved for the treatment of trypanosomiasis and onchocerciasis and a known P2 (ATP/UTP purine receptor) antagonist. Here, we report its ability to inhibit the important CD40-CD154 costi-mulatory interaction required for T cell activation and the development of an effective immune response. In vitro, it inhibited the binding of both human and murine CD154 (CD40L) to their receptor (CD40) even in the presence of protein-containing media and prevented the CD154-induced proliferation of human B cells as well as the corresponding increase in surface expression of CD86, CD80, CD40, and MHC class II in a concentration-dependent manner. Furthermore, in isolated human islets, it also decreased the CD154 induced release of inflammatory cytokines such as IFN-g, interleukin-6 (IL-6), and IL-8. Suramin was selected for investigation because it has been reported to be an inhibitor of the interaction of TNF-a with its receptor and CD154 is a member of the TNF-family. However, it turned out to be a considerably, about 30 fold, more effective inhibitor of the CD40-CD154 protein-protein interaction than of the corresponding TNF interaction. Its median inhibitory concentration (IC50 50 mM) is somewhat higher than for the P2-receptor, but well within the range of its therapeutic concentration levels. Suramin shows considerable polypharmacology, but its interference with the positive costimulatory interaction might provide a possible, not yet identified mechanism for its ability to suppress T cell activity and induce immunosuppression, which might also have limited its clinical usefulness in the treatment of AIDS and cancer. PMID- 19283896 TI - Gastro-intestinal symptoms associated with intense Schistosoma mansoni infection affect class-attentiveness of schoolchildren in Ethiopia. AB - OBJECTIVE: To assess effect of subtle morbidities related to Schistosomiasis mansoni on class-attentiveness of schoolchildren. METHODS: A cross-sectional study was undertaken involving 568 schoolchildren (mean age 13.4 years; 333 from Schistosoma mansoni endemic villages (Sille and Worke-Mado) and 235 from the non endemic town Sheno. A questionnaire on signs and symptoms of ill-health was administered to all study subjects. In addition, parasitological and nutritional anthropometric data were collected. All study subjects from S. mansoni endemic areas were treated with praziquantel and albendazol while children from the non endemic town were only treated with albendazol.FindingsPrevalence of S. mansoni infection was 95% in Sille and 90.6% in Worke-Mado. Overall, among schoolchildren from S. mansoni endemic areas, 36.3% were under-nourished; 66.4% reported easy fatigue/listlessness; 61.3% had abdominal cramps with bloody-mucoid stools, and 17.7% reported to have severe and frequent abdominal cramps which distracted their class-attentiveness. The latter two symptoms were significantly associated with intensity of S. mansoni infection. Among schoolchildren from the non-endemic town Sheno, only 8.9% were under-nourished, 20% reported infrequent abdominal cramps and none had symptoms that were severe enough to affect class attentiveness. CONCLUSION: Symptoms associated with intense S. mansoni infection may negatively affect class-attentiveness of schoolchildren which may have important implications on their overall educational achievements. Further detailed intervention-based prospective studies are recommended to validate these findings which, if confirmed, could further justify school-age targeted praziquantel-based control of schistosomiasis. PMID- 19283895 TI - Oleanolic acid activates Nrf2 and protects from acetaminophen hepatotoxicity via Nrf2-dependent and Nrf2-independent processes. AB - Oleanolic acid is a plant-derived triterpenoid, which protects against various hepatotoxicants in rodents. In order to determine whether oleanolic acid activates nuclear factor erythroid 2-related factor 2 (Nrf2), a transcription factor known to induce various antioxidant and cytoprotective genes, wild-type and Nrf2-null mice were treated with oleanolic acid (90 mg/kg, i.p.) once daily for 3 days. Oleanolic acid increased nuclear accumulation of Nrf2 in wild-type but not Nrf2-null mice, as determined by Western blot and immunofluorescence. Oleanolic acid-treated wild-type mice had increased hepatic mRNA expression of the Nrf2 target genes NAD(P)H:quinone oxidoreductase 1 (Nqo1); glutamate-cysteine ligase, catalytic subunit (Gclc); heme oxygenase-1 (Ho-1); as well as Nrf2 itself. In addition, oleanolic acid increased protein expression and enzyme activity of the prototypical Nrf2 target gene, Nqo1, in wild-type, but not in Nrf2-null mice. Oleanolic acid protected against acetaminophen hepatotoxicity in wild-type mice but to a lesser extent in Nrf2-null mice. Oleanolic acid-mediated Nrf2-independent protection from acetaminophen is, in part, due to induction of Nrf2-independent cytoprotective genes, such as metallothionein. Collectively, the present study demonstrates that oleanolic acid facilitates Nrf2 nuclear accumulation, causing induction of Nrf2-dependent genes, which contributes to protection from acetaminophen hepatotoxicity. PMID- 19283897 TI - Karyotype variability in KP1(+) and KP1(-) strains of Trypanosoma rangeli isolated in Brazil and Colombia. AB - In the present study, the molecular karyotypes of 12 KP1(+) and KP1(-) Trypanosoma rangeli strains were determined and 10 different molecular markers were hybridized to the chromosomes of the parasite, including seven obtained from T. rangeli [ubiquitin hydrolase (UH), a predicted serine/threonine protein kinase (STK), hexose transporter, hypothetical protein, three anonymous sequences] and three from Trypanosoma cruzi [ubiquitin-conjugating enzyme E2 (UBE2), ribosomal RNA methyltransferase (rRNAmtr), proteasome non-ATPase regulatory subunit 6 (PSMD6)]. Despite intraspecific variation, analysis of the karyotype profiles permitted the division of the T. rangeli strains into two groups coinciding with the KP1(+) and KP1(-) genotypes. Southern blot hybridization showed that, except for the hexose transporter probe, all other probes produced distinct patterns able to differentiate the KP1(+) and KP1(-) genotypes. The UH, STK and An-1A04 probes exclusively hybridized to the chromosomes of KP1(+) strains and can be used as markers of this group. In addition, the UBE2, rRNAmtr and PSMD6 markers, which are present in a conserved region in all trypanosomatid species sequenced so far, co-hybridized to the same T. rangeli chromosomal bands, suggesting the occurrence of gene synteny in these species. The finding of distinct molecular karyotypes in KP1(+) and KP1(-) strains of T. rangeli is noteworthy and might be used as a new approach to the study of genetic variability in this parasite. Together with the Southern blot hybridization results, these findings demonstrate that differences at the kDNA level might be associated with variations in nuclear DNA. PMID- 19283898 TI - Unique behavior of Trypanosoma dionisii interacting with mammalian cells: invasion, intracellular growth, and nuclear localization. AB - The phylogenetic proximity between Trypanosoma cruzi and Trypanosoma (Schizotrypanum) dionisii suggests that these parasites might explore similar strategies to complete their life cycles. T. cruzi is the etiological agent of the life-threatening Chagas' disease, whereas T. dionisii is a bat trypanosome and probably not capable of infecting humans. Here we sought to compare mammalian cell invasion and intracellular traffic of both trypanosomes and determine the differences and similarities in this process. The results presented demonstrate that T. dionisii is highly infective in vitro, particularly when the infection process occurs without serum and that the invasion is similarly affected by agents known to interfere with T. cruzi invasion process. Our results indicate that the formation of lysosomal-enriched compartments is part of a cell-invasion mechanism retained by related trypanosomatids, and that residence and further escape from a lysosomal compartment may be a common requisite for successful infection. During intracellular growth, parasites share a few epitopes with T. cruzi amastigotes and trypomastigotes. Unexpectedly, in heavily infected cells, amastigotes and trypomastigotes were found inside the host cell nucleus. These findings suggest that T. dionisii, although sharing some features in host cell invasion with T. cruzi, has unique behaviors that deserve to be further explored. PMID- 19283899 TI - Novel point mutations in sulfadoxine resistance genes of Plasmodium falciparum from India. AB - Point mutations in the dhfr and dhps genes of Plasmodium falciparum are associated with pyrimethamine and sulfadoxine resistance respectively. In this study we have analyzed these genes from Bikaner (situated in North-West region of India), where both uncomplicated and severe manifestations of P. falciparum malaria are seen. A majority of isolates showed double mutant allele for DHFR. In contrast, the only reported mutation present in DHPS was A437G in few isolates. In addition, three novel non-synonymous mutations were observed in the PfDHPS from this region viz., S587F, N666K and C668W. The mutations at the 666 and 668 codon seem to form a bend in the big loop region of the DHPS enzyme and may affect the binding of the drug to the enzyme. Molecular docking of sulfadoxine to this mutated structure indicates reduction in its binding affinity to this enzyme. PMID- 19283901 TI - A review of the laser's role in periodontal practice. PMID- 19283900 TI - Identification of genetic markers in sodium antimony gluconate (SAG) sensitive and resistant Indian clinical isolates of Leishmania donovani through amplified fragment length polymorphism (AFLP). AB - Sodium Antimony Gluconate (SAG) is currently used worldwide as the first-line drugs for the treatment of visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) since 1940s. Unfortunately, the resistance of Leishmania parasite to this drug is increasing in several parts of the world. The mechanism of drug resistance in clinical isolates is still not very clear. Earlier, we have established a differentiation between six clinical isolates as sensitive and resistant on the basis of their sensitivity to SAG in vitro and in vivo as well as expression of proteophosphoglycan contents. In this preliminary study, we have further analyzed these isolates on the basis of their genetic diversity, molecular variance and phylogenetic structure using for the first time, a fingerprinting approach--amplified fragment length polymorphism (AFLP). Altogether 2338 informative AFLP bands were generated using 10 selective primer combinations. Percentage of polymorphism was 55.35%. A number of unique AFLP markers (217) were also identified in these strains. It was deduced that a higher rate of variations occurred among Leishmania clinical isolates which indicate the shifting of drug sensitive nature of parasite towards resistant condition. PMID- 19283902 TI - Notice of retraction. PMID- 19283903 TI - [First experience with the application of rituximab for the treatment of patients with Sjogren's syndrome and disease]. AB - Efficiency and tolerability of rituximab therapy were assessed in 13 patients with Sjogren's syndrome and disease (10) (SLE-1, RA-2). Nine patients (SD-8, PA 1) presented with lymphomas and 4 with systemic manifestations of the disease. Complete and partial remission of lymphoma was achieved in 7 (78%) and 2 (22%) patients respectively. Beneficial effect of therapy on systemic manifestations of the disease was recorded in 3 (75%) of the 4 cases and only 1 patient had cryoglubulinemic glomerulonephritis resistant to rutiximab. Subjective improvement of glandular manifestations was reported by 12 (92%) patients. Objective improvement of salivation and lacrimation was documented in 7 (54%) and 6 (48%) patients who had residual secretion before therapy. Positive outcome of therapy in 12 patients was associated with complete depletion of CD20+ lymphocytes in peripheral blood. These cells were found in a patient who died despite the treatment. Rutiximab significantly decreased medians of ESR, IgG, IgA, IgM, gamma-globulin, and RF levels (p = 0.05-0.002). In 8 patients, therapy resulted in the disappearance of blood cryoglobulins. Intravenous premedication with 500 mg methylprednisolone prevented side effects of rutiximab. The study showed high efficiency and good tolerability of rituximab in combination with pulsed therapy with alkylating cytostatics and courses of polychemotherapy for the treatment of lymphoma and cryoglobuliemic vasculitis in patients with Sjorgen's syndrome and disease. Combination of rutiximab and pulsed therapy was more efficient than rutiximab monotherpy in patients with grade I-IIE MALT-type lymphomas. Rutiximab decreased systemic and glandular manifestations of Sjogren's disease and syndrome in 75 and 48-54% of the patients respectively. PMID- 19283904 TI - [A method for airtight sealing of bronchial stump]. AB - A new technique for airtight sealing of bronchial stump was employed for the first time in 5 patients with fibrocavemrnous tuberculosis complicated by pleural empyema. The mechanical stitch was covered with plastic material on a pedicle from a membranous part of the bronchus; it was buried and sealed with the help of penetrating interrupted sutures. Five patients with similar forms of pulmonary tuberculosis comprised the control group. The new method proved to be highly efficient, promoted early healing of the bronchus, and reduced the development ofbronchial fistulas especially in cases complicated by pleural empyema. PMID- 19283905 TI - [Problems and prospects of experimental modeling of hypoxy-ischemic lesions in the central nervous system]. AB - Perinatal hypoxy-ischemic brain lesions are one of the main causes of mortality and dysfunction of the central nervous system in the neonatal period accounting for high disability rate among survivors. Numerous animal models were proposed to study this problem in ante-, intra-, and neonatal periods of ontogenesis. This paper is devoted to the analysis of the adequacy of these models. The processes of brain development in laboratory animals are considered along with etiopathogenetic factors that can be reproduced on the models of perinatal hypoxy ischemic lesions in CNS. The available data on such models and their correspondence to known clinical syndromes are summarized. Current trends in the development of new models of hypoxy-ischemic brain lesions are discussed. PMID- 19283906 TI - [The use of statins, a new approach to the treatment of autoimmune diseases]. AB - HMG-CoA inhibitors (statins) are widely used for the prevention of cardiovascular events and the management of hypercholesterolemia. Recently, multiple cholesterol independent properties of statins have been discovered. The present review focuses on immunomodulatory and anti-inflammatory effects of statins and their implications for the treatment of autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, and psoriasis. PMID- 19283907 TI - [Somatic mosaicism in mice detected using the RAPD-PCR method]. AB - 99 DNA samples of organs and tissues of 18 mice were examined using the method of PCR amplification with random primers. Among 27 oligonucleotide primers tested, 4 producing stable, well-reproducible profiles of amplification products were chosen for further analysis. Using 2 of these primers we detected differences in RAPD-profiles in some tissues in several individuals. These differences were associated with the modification of mobility, or with the fragment gain/loss in the RAPD profile, and could be caused either by genomic rearrangements, or mutations involving the regions of the DNA-primer pairing. Different epigenetic factors may also contribute to this process. PMID- 19283908 TI - [Synthesis of N-acyl-homoserine lactones, phenazines, some enzymatic activities and antifungal activity of Pseudomonas chlororaphis 449 cells carrying an inactivated rpoS gene]. AB - Inactivation of the rpoS gene encoding for sigma S subunit of RNA polymerase of the rhizospheric strain Pseudomonas chlororaphis 449 results in a sharp decrease (5-8 fold) of phenazine antibiotics synthesis, decline of acid and alkaline phosphatases (pH 2.5 and 8.8, respectively) activities and antagonistic activity of this strain against phytopathogenic fungus Rhizoctonia solani. A mutation in the rpoS gene causes a small decrease of lipase and proteolytic activities in supernatants of Pseudomonas chlororaphis 449 cultures, as well as does not substantially affect the synthesis of three types of N-acyl-homoserinelactones that are signal molecules of Quorum Sensing regulation, and the capacity of bacteria to motility on the surface of the medium (swarming). PMID- 19283909 TI - Re: Ocular side-effects of urological pharmacy. PMID- 19283910 TI - The range of molecular methods for typing Malassezia. AB - PURPOSE OF REVIEW: The recent sequencing of the whole genome of Malassezia globosa and M. restricta forms the basis for molecular epidemiology studies and instigates investigations into their respective virulence factors. Thus, reviewing current knowledge on Malassezia molecular typing methods would reveal the pros and cons of each method and would highlight potential scarcity of epidemiological data regarding this ubiquitous fungal commensal and pathogen. RECENT FINDINGS: Methods employed for Malassezia molecular typing can be categorized into those detecting sequence variations of strains and those that selectively amplify polymorphic DNA markers for discriminating Malassezia species subtypes. The former exploit rRNA gene sequence variations in order to trace M. globosa, M. restricta and M.pachydermatis subtypes associated with specific skin diseases, or detect M. furfur geographical variations. Polymorphic DNA amplification methods, such as amplified fragment length polymorphism analysis, demonstrated association of M. furfur subtypes with the origin of the strain (skin or systemic isolate), whereas PCR-fingerprinting of the mini-satellite DNA clustered M. furfur strains according to their geographic origin and disease origin. Moreover, much typing work has already been performed regarding the zoophilic species M. pachydermatis and the relevant methods can be adapted for studying the anthropophilic Malassezia species. SUMMARY: In the near future, molecular typing will be a powerful tool in epidemiological studies that could be employed for the elucidation of the pathobiology of Malassezia species in associated skin diseases PMID- 19283911 TI - Current treatments for pediculosis capitis. AB - PURPOSE OF REVIEW: Following the increase in prevalence of head lice during the 1990s, research interest in human louse infestation is now greater than at any time since World War I. Problems with treatment, resulting from the selection of populations of lice resistant to insecticides by overuse of some types of product, have triggered an interest in the development of novel therapeutic agents. RECENT FINDINGS: This review first discusses the background to treatment options, basic diagnostic criteria for deciding whether treatment is required and the efficiency of different techniques. Following recent evidence for resistance to insecticides, alternative therapies are examined, including plant-derived essential and fixed oils that are seen by consumers as safer alternatives to pesticides, physically acting preparations and combing options. SUMMARY: I have found that most diagnostic methods lack evidence of efficacy or effectiveness and the evidence for efficacy of several 'popular' options for alternatives to insecticides in treatment is either scant or missing. Claims related to the activity of some products need to be examined more deeply to determine whether they are of real value. PMID- 19283912 TI - Extensively drug-resistant tuberculosis. AB - PURPOSE OF REVIEW: To describe the origin, epidemiology, diagnosis, treatment, prevention, and control of extensively drug-resistant tuberculosis (XDR TB). RECENT FINDINGS: XDR TB is defined as the occurrence of TB in persons whose Mycobacterium tuberculosis isolates are resistant to isoniazid and rifampin and to any fluoroquinolone and at least one of three injectable second-line drugs (i.e., amikacin, kanamycin, or capreomycin). As of June 2008, XDR TB has been found in 49 countries including the United States. It generally takes several weeks to detect XDR TB using conventional culture-based methods, although some progress is being made in developing rapid molecular tests. Treatment for XDR TB is difficult, usually requiring at least 18-24 months of four to six second-line anti-TB drugs. Treatment success rates are generally 30-50%, with very poor outcomes in HIV-infected patients. Management of contacts to infectious XDR TB patients is complicated by the lack of a proven effective treatment for XDR latent tuberculosis infection. SUMMARY: XDR TB is an emerging global health threat. The disease is difficult and expensive to diagnose and treat, and outcomes are frequently poor. New rapid diagnostic tests and new classes of anti TB drugs are needed to successfully combat this global problem. PMID- 19283914 TI - Should HIV therapy be started at a CD4 cell count above 350 cells/microl in asymptomatic HIV-1-infected patients? AB - PURPOSE OF REVIEW: The aim is to review the available data that contribute to the debate on the optimal time to initiate highly active antiretroviral therapy (HAART) in HIV-infected individuals with a CD4 cell count more than 350 cells/microl. RECENT FINDINGS: Although few randomized data exist that can contribute to this debate, a number of findings from observational studies generally support earlier initiation of HAART. In particular, the findings that death rates remain higher in HIV-infected individuals than in uninfected individuals, even when successfully treated, and that both AIDS and several serious non-AIDS events are more common in those with a lower CD4 cell count (even when this count is above 350 cells/microl), suggest that earlier initiation of HAART may prevent much of the excess morbidity and mortality that remains in this patient group. SUMMARY: Currently, the data would generally support initiation of HAART in patients with CD4 cell counts more than 350 cells/microl. However, given the strong potential for confounding in observational studies and the lack of adjustment for lead-time bias in many analyses, it is not possible to rule out possible long-term detrimental effects of earlier use of HAART until the results from fully powered randomized trials that directly address this issue become available. PMID- 19283913 TI - New diagnostic methods for tuberculosis. AB - PURPOSE OF REVIEW: During the last decade, laboratory tests for the detection of Mycobacterium tuberculosis (Mtb) have improved dramatically. Improvements in the ability to detect latent infection with Mtb, disease associated with Mtb, and strains resistant to commonly used antibiotics are reviewed. RECENT FINDINGS: Advances in the detection of Mtb include light-emitting diode fluorescence microscopy, nucleic acid amplification of Mtb and drug-resistant strains, and more rapid liquid culture with adjunct drug susceptibility testing. In the detection of latent tuberculosis infection, interferon [gamma] release assays offer improved accuracy over the tuberculin skin test. SUMMARY: The past 10 years have seen the most rapid growth in new diagnostics for Mtb in over a century. Although these tests offer improvements in the ability to detect Mtb, drug resistant isolates, and those with latent tuberculosis infection, these improvements are counter-balanced by the need to deploy these tests in areas where Mtb burden is highest. PMID- 19283916 TI - The role of radiopharmaceuticals in drug discovery and development. PMID- 19283915 TI - Health-related quality of life in outpatients with COPD in daily practice: the VICE Spanish Study. AB - BACKGROUND: The objective of this study was to measure health-related quality of life (HRQL) in outpatients with chronic obstructive pulmonary disease (COPD) and to assess differences in HRQL according to age, gender, and severity of COPD. METHODS: A total of 9405 patients (79% men, mean age 68 years) participated in a cross-sectional study. HRQL was measured with the Short Form 12 Health Survey Questionnaire (SF-12). Severity of COPD was graded into three levels according to forced expiratory volume in one second value. RESULTS: COPD severity was mild in 33.8% of cases, moderate in 49.3% and severe in 16.8%. The mean physical component summary (PCS-12) and mental component summary (MCS-12) scores were 36.8 +/- 10.4 and 47.2 +/- 11.2, respectively. General health and physical functioning domains were those with the lowest scores. The mean MCS-12 scores were significantly higher in men (47.9 +/- 10.9) than in women (44.1 +/- 11.8) (P < 0.001). Patients older than 60 years rated HRQL worse than patients aged 40-59 years. There were statistically significant differences according to severity of disease in the mean scores of all domains of the PCS-12 and MCS-12 scales. CONCLUSIONS: The present findings show the influence of female gender, older age and moderate-to-severe of airflow limitation on HRQL in outpatients with COPD attended in daily practice. PMID- 19283917 TI - Societal attitudes regarding GM food: the case of Poland within the European Union. PMID- 19283918 TI - Ovarian suppression/ablation in premenopausal ER-positive breast cancer patients. Issues and recommendations. AB - Endocrine therapy remains pivotal in the adjuvant therapy of premenopausal women with hormone receptor-positive breast cancer. Ovarian ablation, used alone, is effective in delaying recurrence and increasing survival in such women. When added to chemotherapy, it is less clear that this technique is effective, perhaps because of the endocrine ablative effect of chemotherapy. Adjuvant trials comparing ovarian ablation with or without tamoxifen to CMF-type chemotherapy (cyclophosphamide, methotrexate, fluorouracil) suggest that the endocrine therapy is equivalent to or better than this chemotherapy in women whose tumors express estrogen and/or progesterone receptors. Endocrine therapy with ovarian ablation, tamoxifen, or the combination is also useful in the metastatic setting in premenopausal women. PMID- 19283919 TI - Optimizing endocrine therapy in premenopausal ER-positive breast cancer patients. PMID- 19283920 TI - Adjuvant endocrine therapy for premenopausal hormone receptor-positive breast cancer; much done, more to do. PMID- 19283921 TI - Expanding role of ovarian suppression/ablation in premenopausal ER-positive breast cancer: issues and opportunities. PMID- 19283922 TI - Perioperative treatment of gastrointestinal stromal tumors. AB - This review describes the current multidisciplinary management of gastrointestinal stromal tumor (GIST), which is the most common sarcoma of the gastrointestinal tract. Before 2001, surgery was the only effective therapy for GIST. The discovery of the central role of KIT proto-oncogene mutations in the pathogenesis of this tumor, and the development of specific inhibitors of KIT tyrosine kinase (TK) function, has changed the paradigm of treatment for GISTs. Imatinib and sunitinib are TK inhibitors with activity against GISTs. Their major established role in GIST is in the treatment of advanced disease. A growing body of literature and clinical experience support the potential perioperative use of these drugs. The adjuvant use of imatinib is based on retrospective series and limited prospective studies demonstrating that imatinib reduces the risk of recurrence. Ongoing studies are further defining the length of adjuvant therapy, as well as identifying the patients that could achieve the best results. Neoadjuvant treatment often decreases the tumor size, allowing a less morbid surgery, appears to be safe and beneficial for some patients, and therefore deserves further study. PMID- 19283923 TI - Perioperative tyrosine kinase inhibitors for GIST: standard ... or an idea that needs further investigation? PMID- 19283924 TI - Controversies in the surgical management of GIST in the era of imatinib. AB - Because of its relatively low toxicity and significant efficacy in the treatment of GIST, imatinib has dramatically altered the natural history of this disease. Early studies, however, have not adequately addressed the optimal length and dose of adjuvant and neoadjuvant imatinib therapy, defined the subset of candidates most likely to benefit from such therapy, determined the long-term impact on OS and use of second-line therapy, nor evaluated the extent and timing of surgery in patients with metastatic disease. With the expectation that advances in biology will lead to more effective chemotherapy for a variety of solid tumors, lessons learned in GIST management will provide a model for multimodality treatment that can be broadly applied. PMID- 19283925 TI - Pharmacology of antineoplastic medications in older cancer patients. AB - Older patients are more susceptible to the complications of chemotherapy, and may be less equipped to react to these complications. After an introduction to the basic principles of geriatric medicine, this article explores the treatment of older cancer patients with systemic chemotherapy, including discussions of the pharmacology of aging, the effectiveness and toxicity of antineoplastic treatment in this population, and issues that need to be addressed in future clinical trials. PMID- 19283926 TI - Older patients and the shifting focus of cancer care. PMID- 19283927 TI - Pharmacology of anticancer agents in the elderly: the pharmacist's perspective. PMID- 19283928 TI - A patient with metastatic melanoma of the small bowel. AB - We report the case of a 74-year-old man with metastatic melanoma of the small bowel. Melanoma metastasizing to the small bowel is a rare but well described presentation of the disease, detected clinically in only 2% to 5% of these patients. Its presentation is similar to other gastrointestinal tract tumors, with symptoms of abdominal pain or anemia prevailing. Recent studies have implicated the chemokine receptor CCR9 and its ligand CCL25 as signals that allow malignant melanoma cells to preferentially metastasize to the small bowel. Common imaging modalities used to detect these small bowel lesions include contrast enhanced computed tomography (CT) scans and upper gastrointestinal series with small bowel follow-through. Given the low sensitivity of these modalities, newer helical CT scanners, 18F-2-fluoro-2-deoxy-D-glucose-positron emission tomography (FDG PET)/CT, and capsule endoscopy are now being recommended to replace the older imaging techniques. Current treatment modalities include surgical resection, which has been shown to increase overall survival, and adjuvant immunotherapy, whose efficacy is currently being questioned. A review of the current literature describing this rare occurrence is included to compare with our patient's presentation, diagnosis, and management. PMID- 19283929 TI - Aggressive surgical therapy for metastatic disease is appropriate in the current management of melanoma. PMID- 19283930 TI - Complementary therapies, herbs, and other OTC agents. PMID- 19283931 TI - Going green. PMID- 19283932 TI - High blood pressure--when should employees be sent home from work? PMID- 19283933 TI - Sociocultural contexts and worker safety and health: findings of a study with Chinese immigrant restaurant workers. AB - More immigrants are seeking employment in restaurants. Drawing data from an ethnographic study, this article discusses what and how sociocultural contexts shape the safety and health of immigrant restaurant workers. Eighteen Chinese immigrants from China, Hong Kong, and Taiwan participated in the study. Data generation methods included a questionnaire, individual and focus group interviews, and participant observations. Ethnographic analysis revealed that immigration mechanisms, demands of English proficiency for employment, and existence of networks and ethnic communities shaped the participants' employment choices. Working hours and schedules, interpersonal relationships at work, job design and training, occupational safety and health training, and national events and economy further influenced the participants' occupational experiences and well-being. Issues were noted with job security, mental health, family relationships, and risks for occupational injuries and illnesses. Implications for occupational health nursing research and practice to reduce immigrant workers' vulnerability to poor safety and health outcomes conclude this article. PMID- 19283934 TI - Development and evaluation of a mental health care system at a Japanese company. AB - Compared with other industrialized countries, the suicide rate in Japan is high. The purpose of this research was to evaluate the mental health care offered at a newspaper facing organizational restructuring and with an aging work force. The Health and Safety Committee played a central role in the creation and application of a new mental health care system. This plan, developed to meet the on-site dynamics of the company, was based on the Ministry of Health, Labour and Welfare's "Guide for Workers' Mental Health Promotion in the Workplace" Executive officers' roles were defined at the directors' meetings, and the Health and Safety Committee held Listening Skills Seminars for executive personnel. As a result, the mental health care system functioned effectively and fostered the care of two presenting employees (i.e., one with slight depression with chronic lumbago and the other with alcohol dependency). In addition, the number of workers who took 4 or more days of sick leave decreased from 31 of 190 in 1996 to 5 of 108 in 2004, with no psychologically related absences. Moreover, the number of industrial injuries declined from 8 cases in 1996 to 0 cases in 2004, and no occupational deaths or early retirements occurred between the fiscal years of 1996 and 2004. As a result of this research at the company, a worksite where repeated large-scale organizational restructuring has occurred, the mental health care system functioned effectively and contributed to a decrease in the rate of sick leave taken by workers. PMID- 19283935 TI - Worksite wellness: a cholesterol awareness program. AB - A 7-month intervention was undertaken to determine the impact of education and coaching on lifestyle choices and lipid values among employees with hyperlipidemia. Four classes over 2 months at the worksite during work time and two telephone interventions were provided with pre, mid, and post data collection. Total cholesterol and low-density lipoprotein values improved during the intervention. Positive lifestyle changes were made involving exercise and diet. Appropriate physician visits and continuous health care increased. Lipid based interventions at the worksite can elicit positive changes in lifestyle, appropriate health care use, and improved lipid values. PMID- 19283936 TI - Managers' perceptions of the current and future role of occupational health nurses in Australia. AB - Occupational health nurses face competition from other professionals in the field of occupational health and safety. This study investigated managers' perceptions of Australian occupational health nurses' roles. Managers were asked to rate the importance of occupational health nurses' activities and the time they believe occupational health nurses do or should dedicate to each activity now and in the future. The questionnaire included 22 activity statements grouped into eight areas of practice that were thought to constitute the occupational health nurse role, based on the Australian College of Occupational Health Nurses standards. Data revealed that emergent roles focused on injury prevention, health promotion, management, and research were of increasing importance, with more time being needed for them in the future. Fulfilling these expectations may place occupational health nurses in competition with other occupational health and safety personnel for particular responsibilities and may require negotiation to gain support for taking on these role activities. Fulfilling these emergent role activities effectively will require appropriate professional development and advanced education. PMID- 19283937 TI - Workplace incivility, bullying, and mobbing. PMID- 19283938 TI - First, do no harm. PMID- 19283939 TI - Manual patient lifting harms nurses, too. PMID- 19283940 TI - Circumcision care. AB - Though circumcision can be a debatable topic, the American Academy of Pediatrics (AAP) in 1999 determined that scientific evidence demonstrated potential medical benefits of newborn male circumcision. However, there was insufficient data to prove neonatal circumcision is medically essential. The opinion of the AAP was reaffirmed in 2005.1 Therefore, nurses in the postpartum units often address questions about circumcision. Moreover, learning about circumcision's standard of care is a great opportunity to add to evidence-based clinical knowledge for the nursing profession. PMID- 19283941 TI - Treating morbid obesity. PMID- 19283942 TI - Rescuing soldiers. PMID- 19283943 TI - Documentation & litigation. AB - What you see on "CSI" or "Law and order" may give you the idea that medical malpractice attorneys routinely use DNA, voice print analysis, and high-tech laboratory assays to prosecute or defend lawsuits. For the most part, this is not the case. With the exception of whatever witness testimony may be available, the only evidence in a malpractice case is the medical record. The patient's chart is used to demonstrate accreditation and regulatory compliance, and to make reimbursement determinations. It also is examined by licensing boards in deciding disciplinary action. For these reasons, it is imperative that nurses consistently use acceptable documentation practices. PMID- 19283944 TI - Taxes for the self-employed nurse. PMID- 19283945 TI - My mother, our patient. PMID- 19283946 TI - Editors' introduction: HIV and other infectious diseases among drug-involved offenders. PMID- 19283947 TI - Integrating an HIV/HCV brief intervention in prisoner reentry: results of a multisite prospective study. AB - Brief interventions to reduce harmful or problem behaviors have become increasingly popular in a variety of health fields, including HIV and hepatitis risk reduction. A central issue in intervention research involves the evaluation of what constitutes an effective "dose" of an intervention. This research examines the relative effectiveness of three alternative brief interventions of varying intensity designed to change the risk behaviors of inmates who are reentering society: a DVD-based, peer delivered intervention; the NIDA Standard HIV Intervention; and a standard practice condition (HIV educational video). All participants randomly received one of the interventions and were tested for HIV and HCV prior to release from custody. Thirty and ninety-day follow-ups examined changes in high-risk behaviors. Results reported here for 343 subjects who have completed the 90-day follow up indicate significant reductions in reported sexual risk behaviors for those participating the DVD intervention, compared to the other two brief interventions. This study is among the first to report any positive impacts on sexual behaviors among a population of inmates returning to the community. PMID- 19283948 TI - The persistence of HIV risk behaviors among methamphetamine-using offenders. AB - Studies have shown that methamphetamine (MA) is rapidly becoming the drug of choice for a large number of substance-abusing offenders and is associated with significantly higher levels of HIV risk behaviors prior to their incarceration. Despite these findings, there has been little follow-up research to determine whether these patterns persist among recently paroled offenders after attendance in an in-prison treatment program. This study uses the self-reported data from 812 substance-abusing offenders in a multisite NIDA-funded project to determine whether, either before incarceration or nine months after release from an in prison substance abuse program, MA use in the past 30 days was associated with increased HIV risk behaviors. The findings indicate that offenders who used MA prior to and after incarceration and treatment report higher levels of HIV risk behaviors compared with offenders with no MA use. Clinical and policy implications of the findings are discussed. PMID- 19283949 TI - Applying classification and regression tree analysis to identify prisoners with high HIV risk behaviors. AB - Among prisoners, past research has associated several factors with HIV risk behaviors, including illicit drug use, engaging in sex trade, older age (for drug related risk), younger age (for sex-related risk), low education, low income, type of offense, history of abuse, mental health disorders, vulnerability and low self-perceived efficacy. This study employs data collected through the Transitional Case Management study of the Criminal Justice Drug Abuse Treatment Studies collaborative to analyze characteristics of prisoners who engaged in high risk behaviors prior to incarceration. For the first 787 participants of this study, we employed recursive partitioning techniques to better identify groups at varying levels of HIV risk behaviors. Those more likely to engage in risky needle use were White and either unemployed and less likely to justify their behavior, or employed with poor decision making capacity. Risky sexual behavior was associated with a general tendency toward risk-taking or a history of unstable housing. Those engaging in any type of HIV risk behavior were risk-takers in general and were aged 25 to 47 with a history of unstable housing. Recursive partitioning, a technique seldom used previously, offers a useful method for identifying subpopulations at elevated risk for HIV risk behaviors. PMID- 19283950 TI - Substance use, mental health problems, and behavior at risk for HIV: evidence from CJDATS. AB - This study examined the relationships between substance abuse, mental health problems and HIV risk behavior in offenders discharged from prison and referred to substance abuse treatment programs. Data from 34 sites (n = 1,358) in a federally-funded cooperative agreement, the Criminal Justice Drug Abuse Treatment Studies (CJDATS), were analyzed. Among parolees referred to substance abuse treatment, self reports for the six-month period before the arrest resulting in their incarceration revealed frequent problems with both substance use and mental health. HIV risk behavior was operationalized as either (a) unsafe injection drug use, e.g., sharing needles and/or sharing injection equipment, or (b) unsafe sex, e.g., sex without a condom. The findings were that (1) unsafe injection drug use was associated with unsafe sex and vice versa, (2) unsafe sex behavior was related to frequency of drug use, and (3) unsafe sex behavior was related to frequency of alcohol use. In these samples, mental health problems did not have a significant effect on risk behavior, controlling for other variables. Future research should probe this "nonfinding" using standardized diagnostic and symptom measures to provide greater detail on the mental health problems (e.g., age of onset, frequency, and severity of the problem). PMID- 19283951 TI - Partner relationships and HIV risk behaviors among women offenders. AB - The HIV infection rate is increasing among women in general and for female inmates specifically (Maruschak 2004), which makes understanding the correlates of risky sexual behaviors critical for this population. Partner relationships, particularly the extent to which women perceive they have power within the relationship, may be important in modeling risk behaviors. Few studies have considered the association between relationship power and HIV risk behaviors among women offenders. This study examines women's perceptions of their relationships using the Sexual Relationship Power Scale (Pulerwitz, Gortmaker, & DeJong 2000) and NIDA's HIV Risk Behavior Assessment (NIDA 1995). Data were collected from female inmates in four prisons as part of the Reducing Risky Relationships for HIV protocol being conducted through the NIDA's Criminal Justice Drug Abuse Treatment Studies (CJ-DATS) cooperative agreement. Women reported whether they had engaged in five types of unprotected sex in the month prior to incarceration. Logistic regression models of the associations between relationship power and five types of unprotected sex revealed some support for the importance of power as a protective factor in reducing the odds of unprotected sexual behaviors. Implications and findings are presented to add to understanding of partner relationships and HIV risk behaviors. PMID- 19283952 TI - Racial differences in HIV/AIDS discussion strategies and sexual risk behaviors among drug-abusing female criminal offenders. AB - African-American female inmates are disproportionately affected by the human immunodeficiency virus (HIV), with heterosexual contact as the primary mode of transmission. This could be the result of racial differences in the strategies used by women to persuade a potential sexual partner to discuss HIV/AIDS and engage in condom use. Data were collected from 336 female inmates as part of the Reducing Risky Relationships for HIV (RRR-HIV) protocol within the Criminal Justice Drug Abuse Treatment Studies (CJ-DATS) cooperative agreement. Bivariate analyses indicated that African-American drug-using women were more likely than Whites to use the rational, withdrawal, and persistence approaches to discuss HIV/AIDS with a sexual partner. Negative binomial regression models were used to identify which interpersonal discussion strategies were significant correlates of the number of the times White and African-American participants had unprotected vaginal sex in the 30 days before incarceration. Results from the multivariate model indicate that White women who are more likely to use the rational discussion strategy were 15% less likely to engage in unprotected vaginal sex; however, these findings were not replicated in the African-American sample. Findings add to the literature on racial differences in HIV/AIDS discussion strategies and sexual risk behaviors among drug-abusing female criminal offenders. PMID- 19283953 TI - HCV in incarcerated populations: an analysis of gender and criminality on risk. AB - While studies have explored the prevalence and correlates for hepatitis C (HCV) infection in substance-using and incarcerated populations these studies have not examined the attributes of criminal histories for those with HCV infection. This study examines the HCV infection rate as it relates to criminal risk factors using baseline data from a randomized trial of re-entering offenders and examines how these risk factors vary by gender. The HCV-positive population had a longer amount of time in confinement (105 vs. 61 months) than those who tested negative. HCV positive men were more likely to currently be receiving drug treatment than women. Criminal risk was positively associated with HCV infection while controlling for major risk factors (OR 1.25,95% CI: 1.07, 1.46), suggesting that the relationship was not spurious. While criminologists tend to examine risk relative to public safety threats, it appears that the public health needs equally require attention. Policy issues are examined regarding how services can be delivered to treat those with HCV infections within the correctional system and address criminal risk factors. PMID- 19283954 TI - Predicting HIV/STD risk level and substance use disorders among incarcerated adolescents. AB - Incarcerated adolescents are among the most vulnerable groups for STD infection, and substance abuse is prevalent in over half of this population. Substance abuse and HIV/STD-associated risk behaviors are closely linked among juvenile justice involved youth, but it is unclear whether common antecedents explain these different problems. The current study examined predictors of HIV/STD risk level and substance use disorders, and investigated whether family variables added unique predictive variance for these problems among incarcerated youth. The sample included 154 substance-involved youth ages 13 to 17 recruited in detention facilities in Miami and Tampa, FL and was primarily male (82%) and African American (58%). Using a comprehensive assessment strategy with data obtained from youth report, parent report, and laboratory confirmed STD testing, the results show that delinquency is a consistent predictor of both HIV/STD risk level and substance use disorders, and also that substance use directly predicts HIV/STD risk level among incarcerated adolescents. Consistent with previous research, family conflict is an important predictor of substance use disorders even after controlling for other factors. The results suggest the need for integrated family based interventions addressing delinquency, substance abuse, and HIV/STD associated risk factors with juvenile justice-involved adolescents. PMID- 19283955 TI - Arrest histories of high-risk gay and bisexual men in Miami: unexpected additional evidence for syndemic theory. AB - Gay and bisexual men continue to suffer the highest burden of HIV/AIDS in the U.S. Since the beginning of the epidemic, substance abuse has been shown to be one of the strongest predictors of sexual risk behaviors and seroconversion among this population. Recent research has focused on additional aspects of health risk disparities among gay and bisexual men, including depression and other mental health problems, childhood sexual abuse, and adult victimization, suggesting that these men are impacted by a syndemic of health risks. The involvement of gay and bisexual men with the criminal justice system is largely absent from the literature. This article describes the nature, extent and predictors of the arrest histories of a sample of gay and bisexual substance users at very high risk for HIV infection and/or transmission. These histories are surprisingly extensive, and are strongly associated with poverty, severe mental distress, substance abuse and dependence, and victimization. The involvement of gay and bisexual men in the criminal justice system deserves a stronger research focus because of the unique challenges facing such men and also because arrests are yet another marker for a host of health risks among them. PMID- 19283956 TI - Beyond spaghetti and meatballs: skin diseases associated with the Malassezia yeasts. AB - Malassezia are common lipid-dependent fungi that grow on the sebaceous areas of human skin, including the face, scalp, and upper trunk. Although Malassezia are a part of the normal human skin flora, they may also cause or exacerbate several skin diseases, including tinea versicolor, Pityrosporum folliculitis, and seborrheic dermatitis. Topical antifungals are the mainstay of treating Malassezia-related diseases. Chronic prophylaxis is often required to prevent recurrences. PMID- 19283957 TI - Evidence-based practice guidelines for skin cancer screening. AB - What does evidence-based practice mean to the dermatology nurse? Specifically, what does the evidence reveal about skin cancer screening? These questions are addressed to help clarify some of the misunderstandings regarding evidence-based practice. PMID- 19283958 TI - Vitamin D: a rapid review. AB - Interest in all aspects of vitamin D seems to be surging due to perhaps the increased number of diverse positive studies suggesting it could prevent a variety of chronic diseases. However, before patients and health care professionals are educated on the preventive aspects of this vitamin that acts more like a hormone, a basic rapid review of vitamin D is needed. There are multiple reasons for the high rate of vitamin D deficiency around the world, including an aging population, obesity, protective skin care measures, skin pigmentation, increased awareness, more utilized diagnostic assays, and perhaps even the lack of natural and fortified food and beverage sources. Various benefits and limitations of vitamin D2 and vitamin D3 supplementation are discussed. The proper use of the vitamin D blood test, also known as "25-OH vitamin D," is important, and changing the normal range of this test may allow for a slightly higher cutoff value based on parathyroid hormone reductions and experience from clinical trials of osteoporosis prevention. The vitamin D doses needed to adequately increase blood levels are provided. Finally, increasing the recommended daily allowance of this vitamin to 800 to 1,000 IU per day may be beneficial for most age groups. PMID- 19283959 TI - What's your assessment? Lichen planopilaris. PMID- 19283960 TI - Concomitant therapy for acne in patients with skin of color: a case-based approach. AB - Acne affects individuals of all races and ethnicities. Patients with skin of color present a unique challenge, particularly due to the potential for post inflammatory hyperpigmentation. Concomitant therapy is an appropriate strategy for this population, as demonstrated in the following case. PMID- 19283961 TI - Bullous pemphigoid. PMID- 19283962 TI - Guidelines for telephone triage. PMID- 19283963 TI - Continued learning builds confidence. PMID- 19283964 TI - Living with melanoma. PMID- 19283965 TI - Leadership: social identity and guiding from within. AB - The precursor to leadership success is the ability to learn about and utilize the social identity of the group as it has evolved. The most successful have expertise in assessing the social identity of groups and developing a commonality of a shared vision that represents the best work of the group and of the leader. PMID- 19283967 TI - New minimally invasive technologies can rejuvenate skin with minimal downtime, less risks. PMID- 19283966 TI - Genetics, melanomas, and teens' indoor tanning. PMID- 19283968 TI - Noncholesterol sterols. AB - Although most of us are more or less familiar with the term "cholesterol", the world of sterols is far more complicated and interesting. Apart from cholesterol, many non-cholesterol sterols can be found in human plasma and these sterols serve many important functions in human organism. They are either derived from endogenous biosynthesis of cholesterol or they come from dietary sources (phytosterols). The sole cholesterol molecule is used for keeping our cell membranes fit, for signalization purposes as well as a precursor for bile acids and steroid hormones. The compounds prior to cholesterol in its biosynthetic pathway were identified as vitamin D3 precursor, meiosis activating sterols and nowadays it seems that they could play a role in cholesterol homeostasis. The sterols from ingested vegetable sources, the phytosterols, are expelled from enterocytes and thus indirectly help our gut in coping with abundant cholesterol in the lumen. Higher plants synthesize many phytosterols, but in marine organisms, we can find other innumerous sterol molecules. The diversity of sterol molecules produced and resistance of their tetracyclic core to enzymatic activities implies crucial importance of sterols during the ontogenesis of multicellular organisms. First oxygen appeared on the Earth app. 2.7 billion years ago and since that time, every new life form took the advantage of oxygen needed also for build-up of sterol molecules. The last decades changed our view to the sterol molecules on almost at all levels of their appearance in human body. In the gut, the absorption of sterols was proven to be protein dependent and the quest for the transporter was successful. The general concepts of intracellular homeostasis of cholesterol have been described including the covalent interaction unbelievable so far - cholesterol and a protein. The clinical importance of non-cholesterol sterols rises with the effort to discover underlying facts about the causes of atherosclerosis. The compound in question, cholesterol, seems to be involved, but it sounds not to be crucial per se. The fact that the accumulation of phytosterols in sitosterolemia enhances the probability of early atherosclerosis onset further supports the hypothesis about some sterol (or steroid) compound being responsible on the molecular level for triggering the pathobiochemical cascade of events leading to atherosclerosis. Understanding the processes taking place in the enterocyte during the absorption of sterols resulted in synthesis of selective inhibitors at the level of sterol translocation into the enterocyte, sterol esterification and chylomicron packing, which are in different phases of clinical testing. The studies in the last part of the monograph represent the clinical potential of the analyses of non cholesterol sterols. In well-defined groups, these analytes enables us to assess the changes in the homeostasis of cholesterol, which can be reflected in the concentration of total cholesterol. Furthermore, the high concentrations of some plasma sterols could point to the inborn errors of cholesterol biosynthesis (Smith-Laemli-Opitz syndrome), transport (sitosterolemia) or metabolization (cerebrotendinous xanthomatosis). Some issues concerning the research on the non cholesterol sterols still remain unanswered - it is not known why some of the enzymes of the cholesterol biosynthesis (seladin-1, sterol D14 reductase) have other functions, qualitative aspects of sterol absorption are not satisfactorily explained and exact reason for expulsion of phytosterols from human body is not clear. Nevertheless, the authors hope that the presented facts can broaden the reader's perspective about the area, which is usually hidden beneath the cholesterol molecule. PMID- 19283969 TI - Dual disorders: nosology, diagnosis, & treatment confusion--chicken or egg? Introduction. PMID- 19283970 TI - Tobacco and psychiatric dual disorders. AB - Smoking is a leading cause of morbidity and premature mortality in the United States. The relationship between tobacco smoking and several forms of cancer, heart disease, stroke, chronic lung disease, and other medical diseases is well recognized and accepted. Recent epidemiological studies are now focusing on the link between tobacco use and psychiatric diseases. Experts now suggest that in the differential diagnosis of "smoker," depression, alcohol dependence, and schizophrenia are highest on the list. Studies are also focusing on the role of secondhand tobacco exposure, either in utero or during childhood, in the risk of dual disorders. Prenatal exposure may alter gene expression and change the risk for a variety of life-long psychiatric diseases, e.g., ADD/ADHD, antisocial personality disorders, substance use disorders, and major depression. Considerable time and effort have been devoted to studying the link between smoking and depression and also schizophrenia. We will focus on less well-studied areas in tobacco use and psychiatric dual disorders (including eating disorders), prenatal and early childhood secondhand smoke (SHS) exposure, and the relationship to the genesis of these dual disorders. PMID- 19283971 TI - Comorbidity and "self-medication". AB - Comorbidity is the rule rather than the exception for substance use disorders, especially when they involve drugs other than alcohol. A common reaction of physicians to comorbidity is to attribute the substance use of addicted people to "self-medicating" with drugs of abuse. When an individual uses an abused drug nonmedically, he or she is seeking brain reward, not treatment. To confuse this drug-using behavior with treatment is to misunderstand addictive behavior and to encourage a false sense that the primary clinical task is to lower the dysphoria associated with the comorbid condition in order to stop the nonmedical drug use. Even the most effective treatment of comorbid disorders is unlikely to halt the addictive drug use. The best way to handle comorbidity (e.g., depression and alcoholism) is to consider each of the comorbid conditions to be a separate and serious illness deserving of effective and specific treatment and not to consider one to be a "symptom" of the other. While it is desirable to treat all comorbid conditions using the best practices for the treatment of that condition it is not reasonable to assume that one of the comorbid disorders is secondary to the other and therefore that treatment of the "secondary" disorder is irrelevant or unnecessary. PMID- 19283973 TI - Weight gain during substance abuse treatment: the dual problem of addiction and overeating in an adolescent population. AB - Obesity and substance abuse during adolescence have reached epidemic proportions, and both are among the leading major public health problems in the United States. There is a significant amount of weight and Body Mass Index (BMI) gain in adolescent ex-addicts during supervised and confirmed abstinence from drugs and alcohol. The primary purpose of this secondary data analysis was to examine the effectiveness of two interventions implemented to address weight management in residential facilities treating adolescent substance use disorders. The secondary purpose was to establish if the outcome was a function of mandated smoking cessation and prescribed psychotropic medications. The results of the study suggest adolescents experienced weight gain in all groups, however there was no interaction effect for smokers and those adolescents on psychotropic medication for either outcome variable. PMID- 19283974 TI - The role of the TCI-R (Temperament Character Inventory) in individualized treatment plannning in a population of addicted professionals. AB - This article discusses a multi-faceted treatment paradigm used to service professionals with addictive disorders. Individualized treatment plans are developed based on a multitude of factors including administration of the TCI-R (Temperament Character Inventory), MCMI-III (Millon Clinical Multiaxial Inventory) and the WASI (Wechsler Abbreviated Scale of Intelligence). Treatment goals include specific therapeutic interventions and patient and therapist directed tasks related to enhanced development of the character dimensions of Self-Directedness, Cooperativeness and Self-Transcendence, as measured on Cloninger's TCI-R inventory. PMID- 19283972 TI - Impulsivity, neural deficits, and the addictions: the "oops" factor in relapse. AB - Impulsive behaviors are observed in a wide range of psychiatric disorders, including substance use, bipolar, attention-deficit hyperactivity, antisocial and borderline personality, gambling, and eating disorders. The shared phenotype of impulsivity is thought to significantly contribute to both the etiology and perpetuation of these disorders. In this review, we focus upon the relevance of impulsivity to the addictive disorders, particularly substance use disorders. First, the literature supporting the presence of impulsive behaviors prior to the onset of drug use and addiction is discussed. The relevance of impulsivity to relapse is then presented, with a focus on three distinct neurocognitive constructs: automaticity, response inhibition, and decision making. Automaticity is a quickly occurring relapse process resulting from the learned habits induced by persistent drug use. Addicted persons with response inhibition deficits are unable to suppress these previously reinforced behaviors. Decision-making deficits contribute to relapse through a poorly considered assessment of the consequences of drug use. The brain regions associated with each model of impulsive behavior are described, and relevant neurobiologic disruptions in addicted subjects are discussed in the context of their specific neurocognitive deficit(s). Descriptive confusions in the terminology and confounds inherent in the study of impulsivity are described. Empirical investigations documenting the hypothesized relationship between specific deficits in impulsive behaviors, coupled with their neurobiological correlates, and relapse should be the focus of future studies. PMID- 19283975 TI - Opiate prescription medication dependence and pain perceptions. AB - Our study showed that the perception of pain lessens with detoxification from chronic prescription opiate medications. Thus, removal of opiates resulted in less pain, and chronic administration of opiates actually increased pain perceptions. The underlying pathophysiology of increased pain sensitivity from chronic administration is not well understood. However, the enhanced pain from the development of tolerance and dependence is a common phenomenon. Clinicians are advised to limit their prescribing of opiate drugs to patients on a chronic basis. Further, unresolved pain complaints, and continued complaints of pain despite escalating doses of opiate medications suggest addiction and its adverse consequences. Identification of addiction and detoxification is the proper approach to pain management in chronic opiate administration. PMID- 19283976 TI - ECG of the month. Variations on the common theme of premature atrial complexes. Sinus rhythm with premature atrial complexes (PACs) occurring singly, in couplets, and in triplets. PMID- 19283977 TI - A 61-year-old man with asymptomatic, bilateral lung masses. PMID- 19283978 TI - A case of persistent shin pain. Longitudinal stress fracture secondary to shin splints. PMID- 19283979 TI - CNS germinoma of the pituitary and pineal regions, lateral ventricle, and fourth ventricle presenting in adulthood. PMID- 19283980 TI - Agnathia-otocephaly. PMID- 19283981 TI - Achievement of glycemic outcome goals in pediatric diabetes: the impact of staff number. AB - Achieving optimal glycemic control has become a priority of diabetes management to minimize complications. However, the impact of staff number on glycemic outcome has not been examined in pediatric diabetes practice. We retrospectively analyzed this relationship in patients attending a pediatric diabetes outpatient clinic from 1997 through 2004. We also examined the impact of the program on hospitalizations. A linear trend for higher staff number was associated with lower HbAlc. The data suggests that a mean clinic HbAlc of 8% might be achieved with 3.2 staff per 100 patients. Repeat hospitalizations were considerably reduced by the program. The preliminary data suggest that an increase in staff number can improve glycemic control and reduce hospitalizations in a largely indigent pediatric diabetes population. We hope that this data will serve as an impetus to further research of workforce requirements to optimize staff number, training, and specialization needed for optimal health outcomes in children with diabetes. PMID- 19283982 TI - Helminthic eosinophilic meningitis: emerging zoonotic diseases in the South. AB - Today most emerging infectious diseases, such as West Nile virus and severe acute respiratory syndrome (SARS), arise in the natural environment as zoonoses and are often imported into the United States (US). The most common helminthic infections that can cause eosinophilic meningitis (EoM) in the US, neuroangiostrongyliasis and baylisascariasis, share many of the characteristics of emerging infectious diseases. Neuroangiostrongyliasis, a rodent zoonosis caused by the rat lungworm, Angiostrongylus cantonensis, is now endemic in the US following the importation of infected rats on container ships and African land snails, the parasite's intermediate hosts, as biological controls and exotic pets. Baylisascariasis, a raccoon zoonosis, caused by the raccoon roundworm, Baylisascaris procyonis, has extended its US distribution range from suburban neighborhoods in the northern US to the Southeast and West Coast since the 1980s. Both A. cantonensis and B. procyonis are now enzootic in Louisiana and have caused EoM in humans. This review analyzes scientific articles selected by MEDLINE search, 1966-2008, in order to assess the evolving epidemiology of EoM in the US, and specifically in Louisiana; and to alert Louisiana clinicians to populations at increased risk of helminthic EoM as a result of age, ethnicity, lifestyle, food choices, location of permanent residence, or recent travel in the Americas or Caribbean. Most parasitic diseases causing EoM are no longer confined to tropical countries; they are now endemic in the US and in Louisiana and more cases may be anticipated. PMID- 19283983 TI - Banning smoking on a substance abuse treatment unit: does it deter patients? AB - There is a national trend toward banning smoking in substance abuse facilities. Conventional wisdom considers it unreasonable to ask patients to stop smoking while withdrawing from other substances and is seen as reducing the chances of success. This project examined whether a smoking ban in an inpatient medical detoxification unit would deter patients. Admission logs were used to identify all patients admitted a year before and after the ban on tobacco. Results indicated stability in: the number of admissions; patient demographics; substance use pattern (alcohol abuse predominated); and tobacco use (80.2% vs. 84.0%). Average length of stay decreased a third of a day (P<.05), which did not appear to be due to discharges to other substance abuse units or leaving against medical advice, as these did not change significantly (P>.10). Thus, the ban did not appear to have any notable impact in deterring patients from seeking or staying in treatment. PMID- 19283984 TI - Moments in time. An interview with Dr. Joseph A. Sabatier, Jr. Interview by Sadie D. Wilks. PMID- 19283985 TI - Calculating CVD risk in older patients. PMID- 19283986 TI - Managing Alzheimer's disease in primary care. PMID- 19283987 TI - Physical activity reduces morbidity in older patients. PMID- 19283988 TI - Diagnosing shoulder pain. PMID- 19283989 TI - Industrial diseases. PMID- 19283990 TI - Acne. PMID- 19283993 TI - What nurses need to know about rBGH. PMID- 19283994 TI - [The involvement of non-resident fathers--a review of the current research]. AB - Non-resident fathers have been forming one of the fastest growing social groups during the last decades. Nevertheless, their involvement and role in the development of their separated children have received a comparatively modest attention in research. Little is known about the possibilities and limits of their influence after parental separation. This review is meant to give a general overview of the current scientific literature about the involvement of non resident fathers. Various factors influencing the behaviour of fathers after separation such as the circumstances around the visits, the line-up of families, the role of mothers and general parental conflicts are taken into consideration. Finally, the current research in this field is critically discussed and future research perspectives are formulated. PMID- 19283995 TI - [Effectiveness of parent training for children with ADHD in routine clinical practice]. AB - Studies on the effectiveness of parent trainings, especially for children treated with stimulants, yielded conflicting results. This study investigated the effectiveness of parent training (PT) as a part of routine clinical care. Compared to a waitlist control (n=16) PT-mothers (n=16) reported significantly fewer ADHD-symptoms, better acceptance of their children and a trend to better relationship satisfaction. No differences were found between children treated with or without stimulants. Contrary to the high level of consumer satisfaction fathers didn't report any improvements on all outcome domains. PMID- 19283996 TI - [School-based prevention of antisocial behavior: efficacy of the "Verhaltenstraining fur Schulanfanger"]. AB - In order to test its short- and middle-term impact as well as differential gender effects the universal school-based "Verhaltenstraining fur Schulanfanger", a German prevention program on antisocial behavior for elementary school students, was conducted and evaluated in Luxembourg. In a quasi-experimental setting, nine first grade classes (n=88) were assigned to intervention and control groups. Three waves of data (pretest, posttest and 12-months-follow up) including teacher and student assessments were analysed. As a result intervention effects in terms of significant reductions in oppositional defiant and aggressive behavior as well as an increase in social and emotional competencies according to teachers' assessments at 12-months-follow-up could be observed. Student interviews showed positive short-term effects in terms of increased emotional knowledge and social problem solving competencies at posttest, with girls showing a stronger improvement of their emotional knowledge than boys. PMID- 19283997 TI - [Psychotherapy with physically challenged children and young people: implications of a change of paradigm]. AB - In the light of newer scientific developments the biopsychosocial approach in psychotherapy appears to be in need of a revision. In neurosciences a paradigm change from a linear to a dynamic outlook on development as a self-organizing process guided by interactions with the environment took place under the heading of Neural Plasticity during the last ten years. This implies that the conditions of development for challenged children are not comparable with those of healthy children as the case-examples of children with Spina Bifida indicate. On this background, a pilot project was launched with the goal of determining which forms of psychotherapy are helpful for challenged children. A practically oriented, eclectic approach was developed applying empiric-regulative cycles which promotes the dynamics of self-organizing psychic and physical processes as shown in a presented example of a child experiencing post-lesional plasticity. Thus, psychotherapy is understood as a co-constructive process of reciprocal shaping of relationship; it fosters beneficial organization processes of psychic and physical impact. PMID- 19283998 TI - [Fetal alcohol syndrome--a frequently missed syndrome]. PMID- 19283999 TI - [Incidence of venous catheter associated septicemias after introduction of a new valve connector]. PMID- 19284000 TI - [Early determination of cardiovascular risk]. PMID- 19284001 TI - [Summary of the 5th Tubingen Specialty Congress of Pediatric Nursing 9-11 October, 2008]. PMID- 19284002 TI - [Infection with human immunodeficiency virus (HIV) in childhood]. PMID- 19284003 TI - [Prepartum parent counseling. A general practice example of nursing for optimal nursing care of expectant parents]. PMID- 19284004 TI - [Decubitus ulcer risk scales for premature and newborn infants]. PMID- 19284005 TI - [Naturopathy in nursing. I: Aroma therapy]. PMID- 19284006 TI - Resident duty hours in the U.S.: new directives. PMID- 19284007 TI - The growing challenge of maternal obesity. PMID- 19284008 TI - LTI or GMS card for chronically ill children? Or both? PMID- 19284009 TI - General practitioners knowledge, practice and training requirements in relation to doping in sport. AB - This study examined General Practitioner's (GP) knowledge, practice and training requirements in relation to doping in sport in Ireland. All 2083 GPs on the Irish College of General Practitioners (ICGP) register received a postal questionnaire, yielding a 37% response rate (N=771, 63% male, average age 46.2 +/- 9SD, range 28 74 years). Results revealed that 14% (112) deemed their knowledge of doping agents to be good or very good, 12% (94) had completed specific training modules in doping or sport, and 24% (183) were connected with a specific sport as a team doctor/advisor. Over one in four (28%: 217) had been consulted for advice on doping in Sport, 33% (256) possessed the current list of prohibited substances, and 25% (190) knew of the Irish Sports Council's drug-testing procedures. The current initiatives to discourage doping in sport were felt to be ineffective, and although 92% (716) indicated that GPs had a role to play in the prevention of doping in sport, only 9% (66) felt adequately trained for such a role. There was overwhelming support for further training among GPs, although the most appropriate method of providing training is complex and requires strategic planning. PMID- 19284010 TI - The uses of mercury equipment and products in Irish healthcare. AB - Mercury is a naturally occurring metal, one of the most toxic metals within the food chain and has a number of uses in the Irish health care sector. This paper reports on a mercury survey in Ireland with the purpose to present a picture of mercury use in health settings. The findings show 50% of health settings free of all mercury medical equipment. Mercury spills occur within 30% of health settings, with no training provided in handling mercury equipment, in the management of mercury spills and only 10% of health services have mercury clean up kits. With 50% of mercury waste disposed inappropriately, a number of recommendations are advanced for reducing the amount of mercury in the health settings alongside awareness and education for health professionals. PMID- 19284011 TI - The role of alcohol in deaths presenting to the coroner's service in Cork City and County. AB - A retrospective study was conducted in order to determine the prevalence and concentration of alcohol in post-mortem blood samples sent for toxicological analysis in Cork City and County in 2003 and 2004. Post mortem reports of these deaths were reviewed for the presence or absence of alcohol at the time of autopsy, blood alcohol concentration (BAC) at time of death, age and sex of the decedents. Of samples sent for blood alcohol analysis (BAA), 38.4% were positive for alcohol. Significant differences were found between the proportions of alcohol positive cases by cause of death. Alcohol positive cases were significantly younger (44.3 +/- 17.8 years) than alcohol negative cases (51.9 +/- 19.4 years) and fifty two percent of drivers were positive for alcohol at the time of death. Awareness of the harmful and potentially fatal effects of alcohol should continue to be raised within the community, so as to prevent future fatalities. PMID- 19284012 TI - Meeting the international adolescent physical activity guidelines: a comparison of objectively measured and self-reported physical activity levels. AB - Objectively measured and self-reported physical activity levels were assessed among Irish adolescents (n=28) by heart rate monitor and self-report questionnaire. The purpose was to determine the percentage of Irish adolescents that met international physical activity guidelines using both objective and subjective measurements. More than 11% of subjects met the international moderate intensity physical activity guidelines for adolescents when measured by self report questionnaire. No subjects met this guideline when measured objectively by heart rate monitor. Levels of agreement between the two measurements were poor (k < 0.40). When measured objectively by HR monitor, no adolescent met the international guidelines on sustained vigorous physical activity. Prevalence estimates for compliance with international physical activity guidelines varied according to mode of measurement. We conclude that compared to self-report measures, adolescent physical activity levels are lower than those required for health benefits when measured objectively, and only a minority of young people satisfy the international recommendations. PMID- 19284013 TI - The Transit/Admission Lounge study. AB - In response to persistent overcrowding of Emergency Departments in Ireland, the Department of Health and Health Service Executive provided funding for "Transit Lounge" areas to be built. These lounges were to provide a location for patients to wait in beds pending the availability of a ward bed. This research was performed to assess the impact of such a lounge on the overcrowding of the Emergency Department and on patient outcomes. The time period from the opening of the Transit Lounge was compared with the same period a year earlier. The Transit Lounge delivers a comfortable place for patients to wait. It does not reduce Emergency Department overcrowding and has been associated with an increased time waiting for a ward bed. The solution to overcrowding is the creation of real capacity in the system so that ward beds are available in acute hospitals for the "unscheduled unwell". PMID- 19284014 TI - How to work up a patient with polyneuropathy? AB - Undiagnosed and untreated neuropathy may lead to disability and poor quality of life. Ordering every possible test to find the cause of polyneuropathy can waste time and resources. In this study, we investigated what could be used as a routine neuropathy screen. A retrospective audit of all charts of patients diagnosed to have polyneuropathy by nerve conduction studies from November 2001 to November 2002 were carried out. Demographics, background history, type of neuropathy and investigations done were documented. The charts of 61 patients were audited. 12 patients had a background history of diabetes mellitus. 2 patients had history of alcohol abuse. 23 patients presented with paraesthesia and 33 with weakness of limbs. We found a cause of polyneuropathy in 79% of cases. In most patients with polyneuropathy where a cause can be identified, this can be achieved by the medical history, neurological examination, nerve conduction studies and the baseline blood tests. We suggest a 3-step approach to the diagnostic workup of polyneuropathy. PMID- 19284015 TI - Etonogestrel implant as a contraceptive choice; patient acceptability and adverse effect profile in a general practice setting. AB - The aim of this study was to evaluate the experience of a cohort of patients utilizing the etonogestrel implant (Implanon) in an Irish General Practice setting. This study involved a survey administered as a telephone questionnaire, to a cohort of women (n=75) who opted to use the etonogestrel implant. 53% reported problems with the implant post insertion, the commonest problem being irregular bleeding in 22% cases. Early removals were documented in 28% cases, of which, 29%, were caused by irregular bleeding. Based on this study, it is evident that women have high expectations of the implant, and counselling about what they can expect during use is important in order to avoid unreasonable expectations. This study also demonstrates that the use of the etonogestrel implant is a valuable contraceptive option, which can be successfully delivered in a GP setting, for both patients of the practice as well as patients referred by colleagues locally. PMID- 19284016 TI - Successful outcome of patients with relapsed/refractor Hodgkin lymphoma treated with high dose chemotherapy at the National Adult Bone Marrow Transplant Unit at St. James's Hospital. AB - Patients with Hodgkin lymphoma who relapse or are refractory to first line multi agent chemotherapy can be successfully salvaged with high dose therapy (HDT) and autologous stem cell transplant (ASCT). Twenty-six patients with relapsed or refractory Hodgkin lymphoma have been treated with HDT and ASCT at St James Hospital between 2000 and 2005. At day 100 post HDT-ASCT, 23 patients were in complete remission. This group included all 6 patients transplanted at first relapse, 8 of 9 with advanced disease and 9 of 11 with primary refractory disease. Patients treated in first relapse had the best outcome with an overall and progression free survival of 100% (median, 37 months). Patients with primary refractory disease had the poorest outcome with an overall survival of 76% (median, 28 months). All patients with primary refractory disease responsive to salvage chemotherapy were in remission at a median of 28 months. The presence of chemosensitive disease prior to transplantation was the most important determinant of outcome. PET-CT imaging is useful to assess chemosensititvity prior to HDT and thus predict which patients will do well post HDT-ASCT. No patient died of treatment related toxicity. The outcome of this patient series compares favourably with international figures. PMID- 19284017 TI - Voriconazole in the treatment of allergic bronchopulmonary aspergillosis in cystic fibrosis. AB - Allergic bronchopulmonary aspergillosis (ABPA) can cause a significant clinical deterioration in patients with cystic fibrosis. There is very little research in the current literature with regard to alternatives for treatment, apart from long courses of steroids. We conducted a retrospective review of all our patients with ABPA treated with the antifungal voriconazole and found there was a significant drop in IgE levels post treatment as well as a decrease in steroid dosing. The improvement in FEV was not statistically significant; however there was a very wide variation in pre-treatment levels. PMID- 19284018 TI - Down's syndrome and hip arthropathy. AB - We report a case of a 19 year old male with Down's syndrome who presented with severe functional disability and pain secondary to bilateral hip arthritis. He had longstanding developmental dysplasia of the hip (DDH) that had evolved into end stage arthritis. Following a comprehensive assessment and counselling, he underwent bilateral total hip replacements. At last follow up, he had excellent outcome for function and pain relief. This case highlights the good results achievable with aggressive management of musculo-skeletal conditions (especially hip arthropathy) in those with Down's syndrome and the need for increased recognition of these conditions. PMID- 19284019 TI - Resuscitation of infants at the threshold of viability. PMID- 19284020 TI - Co-operation leads the way. PMID- 19284021 TI - Initiative aims to boost breast screening uptake. Interview by Christian Duffin. PMID- 19284022 TI - Report urges more co-operation to fight malnutrition in the UK. Interview by David Pountney. PMID- 19284023 TI - Wish upon a star. PMID- 19284024 TI - When information must be handled with care. PMID- 19284025 TI - When the going gets tough... PMID- 19284026 TI - The one-stop shop memory service. Interview by Christian Duffin. PMID- 19284027 TI - Shirley Law. PMID- 19284028 TI - Dishing up good practice. Interview by Jennifer Trueland. PMID- 19284029 TI - Degrees of confidence. Interview by Liz Clark. PMID- 19284030 TI - Prescribing for older people. PMID- 19284031 TI - Promoting good health in people aged over 75 in the community. AB - This article examines a project set up in 2004 to promote the health and wellbeing of people aged over 75 living in Caerphilly, south Wales. The multidisciplinary project offered holistic, nurse-led screening in people's homes. It resulted in reduced admissions to hospital for fractured femur, pneumonia and stroke during the period 2004 to 2006. PMID- 19284032 TI - OIE standards and guidelines related to trade and poultry diseases. AB - Recognising how difficult it is for some countries to fully eliminate animal diseases from their territory as a whole or to maintain an animal disease free status in parts of their national territory, the World Organisation for Animal Health (OIE) has introduced the concepts of 'zoning' and 'compartmentalisation' for the purposes of disease control and international trade. Full definitions of these terms are contained in the OIE Terrestrial Animal Health Code. Compartmentalisation is based on a functional separation by biosecurity measures, whereas zoning is based on a geographical separation. In both cases, relevant animal subpopulations should be clearly defined, recognisable and traceable and should be epidemiologically separated from other subpopulations. Veterinary Authorities as well as the private sector have important responsibilities in establishing and maintaining zones and compartments. PMID- 19284033 TI - Field trial for assessment of avian influenza vaccination effectiveness in Indonesia. AB - The aim of this field study was to determine the efficacy of vaccination against highly pathogenic avian influenza (HPAI) virus strain H5N1 in Indonesia. A limited, prototype clinical trial was performed using a standardised treatment group, in which poultry flocks were vaccinated at least twice with a selected H5N1 vaccine, and a control group comprising flocks treated with non-standardised procedures chosen by the farmer. Each group consisted of six flocks comprising either layers or native chickens. Haemagglutination inhibition (HI) antibody levels were determined by regular serum sampling, and outbreak surveillance relied on non-AI-vaccinated sentinel birds. After three vaccinations high antibody titres were produced in the treatment group, and the percentage of layers with an HI titre > 40 was approximately 90%. Although no conclusions can be drawn regarding reduction of virus transmission, this study demonstrated that 11 farms remained free from AI during the observation period, and that a surveillance programme based on differentiating infected from vaccinated animals (DIVA) can be implemented. PMID- 19284034 TI - [Prioritising the risk of foodborne zoonoses using a quantitative approach: application to foodborne bacterial hazards in pork and beef]. AB - Foodborne zoonoses are a major public health concern. Risk analysis, which underpins international policies on food safety and trade in foodstuffs of animal origin, requires that an assessment be made of the occurrence and severity of human cases for each type of foodstuff. However, the tools currently available for quantifying risks are only capable of estimating the consequences of certain diseases. This article proposes an alternative quantitative approach for prioritising the risk of foodborne zoonoses, based on the creation of a typology of hazards and calculating a risk score. A combination of average hospitalisation and mortality rates is used to quantify the severity of human cases. By calculating the percentage of food-associated cases it is possible to estimate the incidence of cases linked specifically with the foodstuff being assessed. This method is illustrated by applying it to bacterial zoonotic hazards in pork and beef and provides a support tool for veterinary public health decision makers. PMID- 19284035 TI - Bacteriological detection of Salmonella enteritidis in eggs: a review. AB - The detection of Salmonella enterica serovar Enteritidis (SE) in eggs is hampered by a typically low prevalence of contaminated eggs, the low number of SE organisms in such eggs, and the presence of inhibitory substances in the egg albumin. For these reasons, the analysis of large pools of eggs is normally necessary, which presents logistic and microbiological challenges associated with a low number of target organisms from a large volume of sample matrix. In some studies using artificially inoculated eggs the standard procedure for Salmonella culture consisting of pre-enrichment, followed by selective enrichment and plating has been replaced by incubation of the egg pools at 25 degrees C to 37 degrees C followed by direct plating. However, in most cases using pools of naturally contaminated eggs, it may be necessary to enhance the traditional three step method by addition of antibiotics or iron supplements. PMID- 19284036 TI - Sampling and bacteriological detection of Salmonella in poultry and poultry premises: a review. AB - The detection of Salmonella in primary poultry production is an issue of great concern in the European Union (EU), since control of this zoonotic disease is in part based on the reduction of the prevalence at the farm level. Success of detection is likely to be highly dependent on the choice of an adequate sampling procedure combined with a sensitive culture method. In poultry farms 'naturally pooled' faeces/litter and dust are the matrices of choice. In floor systems boot swabs are the preferred method for the collection of faeces. A wide range of culture methods is available, but ISO 6579:2002 (Annex D) is currently the standard for poultry environmental samples in the EU. In this review, the authors discuss in detail the range of sampling and culture methodologies for Salmonella in poultry farms. The review also covers sampling and testing of poultry hatcheries, live birds and poultry carcasses. PMID- 19284037 TI - Compartmentalisation and zoning: the Dutch perspective. AB - The aim of the World Organisation for Animal Health (OIE) procedure of compartmentalisation is to contribute to safe trade in live animals and animal products. The fundamental requirement for its application is that the population considered for trade remains epidemiologically separate from populations of higher risk. Compartmentalisation makes use of a functional separation through management, taking into account all relevant epidemiological factors. In this paper, the authors begin by describing current (inter)national developments and actions in this field. Second, some sensitive issues are outlined where one internationally accepted view would help to implement compartmentalisation successfully in international trade. The OIE standards do not contain the procedure for assessing the biosecurity plan, which is crucial. The authors propose the use of a hazard analysis and critical control point system (HACCP) to determine the effectiveness of a biosecurity plan, taking account of all possible risks and potential disease entry points. This could be based on the model of the Codex Alimentarius Commission. Other issues discussed are the outbreak of disease close to a compartment, the role of certification agencies and non-compliance with the biosecurity plan. PMID- 19284038 TI - [Epidemiological surveillance networks for animal diseases in French-speaking West and Central Africa]. AB - One of the objectives of the Pan African Programme for the Control of Epizootics (PACE) was to set up epidemiological surveillance networks in African countries. A survey based on a written questionnaire was conducted to review the technical and institutional organisation of epidemiological surveillance networks in nine French-speaking countries, including five in West Africa (Senegal, Burkina Faso, Cote d'lvoire, Togo and Guinea) and four in Central Africa (Cameroon, Central African Republic, Democratic Republic of Congo and Chad). The survey results showed that there are more similarities than differences among epidemiological surveillance networks. In general, they were found to be technically and institutionally well established. However, the two weaknesses of the majority of networks are the inadequate diagnostic capacity of laboratories and the insufficient operationality of steering committees. Epidemiological surveillance should exclusively be the domain of Veterinary Services and it is crucial for ensuring that any change in the health status of an animal population is detected rapidly. However, the networks' continuing survival after external financing ceases is generally not guaranteed because, in many cases, governments fail to fund them adequately. PMID- 19284039 TI - [Developing a harmonised system for the recognition of clinical trials for veterinary product registration]. AB - The increase of commerce between developing countries requires a harmonised system for accepting the results of clinical trials (CT) of veterinary products, similar to those that exist in developed countries. The objective of this paper is to propose a basis for the creation of a system that harmonises CTs for approving veterinary products (VP) for registration. Such a system would be a step towards unifying the CTs of different countries, while maintaining country specific variations that are compatible with the scientific method, international standards, and the principles of objectivity, transparency and confidentiality. Basic requirements to be fulfilled by both private institutions and public offices are described, as are professional responsibilities and possible administrative procedures that could be adapted in each country. The conclusion reached is that a harmonised system is feasible, as has been demonstrated in numerous countries throughout the world. A harmonised system will result in a more efficient product approval process, a reduction in costs, greater transparency in controls, an improvement in the reliability of the health system, and a reduction in the time the process takes. It will also contribute to animal welfare by avoiding the need to repeat trials. The author acknowledges that there are cultural, technological and economic limitations and that these problems, and others, have yet to be overcome. PMID- 19284040 TI - A comparison of the delivery of veterinary services to small-scale and medium to large-scale poultry keepers in peri-urban Ghana. AB - The study compared assessments of animal health care delivery to small-scale and medium to large-scale poultry keepers in peri-urban areas in four regions of Ghana. Four hundred and one keepers were interviewed using a questionnaire. Significantly higher proportions of the small-scale and medium to large-scale keepers met their needs by themselves. The mean distances or times used in getting veterinary services were similar for both groups. Significantly higher proportions of medium to large-scale keepers said drugs and vaccines were available and getting help was easy. Staff attitude and effectiveness were perceived as good by significantly higher proportions of small-scale keepers. Equity and efficiency were generally poor, while drug costs were considered expensive by medium to large-scale keepers. The study concluded that generally, there were no marked differences in the way in which the two groups perceived the quality of service delivery and the findings did not support claims that small scale keepers receive inadequate services. PMID- 19284041 TI - Client assessment of animal health care delivery in peri-urban Ghana. AB - The study used a questionnaire to assess the delivery of veterinary services as perceived by users in four peri-urban areas in Ghana. Eight hundred and eighty nine respondents were interviewed: 10.7% were cattle farmers, 27.4% were small ruminant farmers, 14.2% were pig farmers, 45.1% were poultry farmers and 2.6% reared various animals on a part-time basis. Most of the animal health needs were either met by the owners (50.4%) or by veterinarians (41.6%). Veterinarians were mainly consulted for advice on animal health, disease diagnosis and treatments. Most respondents (65.7%) had no difficulty in getting help from government services. Higher proportions of interviewees perceived effectiveness, efficiency, service quality, staff attitude and technical competence as 'good' or 'very good'. However, equity and accessibility were thought to be 'fair' to 'very poor', and the cost of drugs was considered expensive' or 'very expensive'. The study identified strengths and weaknesses in the delivery of animal health services in peri-urban Ghana and this information could be used as a basis to improve the overall quality of these services in the future. PMID- 19284042 TI - The transfer of East Coast fever immunisation to veterinary paraprofessionals in Zambia. AB - In eastern Zambia, immunisation by 'infection and treatment' is the main method used to control East Coast fever, an acute and lethal cattle disease. This service, which requires a stringent cold chain, used to be free of charge. When a minimal user fee was introduced, attendance dropped drastically. Consequently, this complex immunisation programme was transferred to veterinary paraprofessionals working on their own account, with the aim of boosting a more sustainable distribution of vaccine. Paraprofessionals were provided with a motorbike and the required specific equipment, but fuel and drugs were at their expenses. The paraprofessionals recovered their costs, with a profit margin, by charging the cattle owners for immunisation. The reasons for the successful transfer of immunisation to paraprofessionals (despite the maintenance of a fee) are attributed mainly to the absence of information asymmetry between the paraprofessional and the livestock owner, the appreciable level of effort of the paraprofessionals and the verifiable outcome of the service provided. PMID- 19284043 TI - A foot and mouth disease simulation exercise involving the five Nordic countries. AB - Simulation exercises are considered a very valuable tool for testing contingency plans established for the control and eradication of rapid spreading animal diseases such as foot and mouth disease, classical swine fever and avian influenza. An inter-Nordic simulation exercise was conducted in 2005 with the objective of testing the national foot and mouth disease contingency plans adopted respectively by Denmark, Finland, Iceland, Norway and Sweden. The Central Veterinary Administrations of the five countries jointly prepared a scenario which involved about 40 livestock holdings, 4 reindeer flocks, 6 slaughterhouses and approximately 500 people. An Excel spreadsheet with information on the events to take place and the timetable to follow during the exercise was a valuable tool for ensuring that the exercise was kept on track. The evaluation of the exercise dealt both with inter-Nordic activities and the activities of individual countries. PMID- 19284044 TI - Economic aspects of foot and mouth disease in Bolivia. AB - The paper presents results from two economic analyses of foot and mouth disease (FMD) in Bolivia. Both recommended a programme to eradicate the disease, but one reported a negative economic return while the later study found a positive and robust return. To investigate the reasons for these differences the paper presents information on cattle movement and how this relates to the epidemiological situation of FMD in Bolivia. This analysis identifies two important trade routes: southern and central cattle trade routes and two FMD endemic areas: the humid tropical areas of the Departments of the Beni and Santa Cruz, and the semi-arid subtropical area to the south east of the country known as the Bolivian Chaco. The farm-level incentives to control FMD in the endemic areas, where cattle are kept in extensive systems, are negative and the main losses caused by the disease occur four to six years after an outbreak. Given this situation it is suggested that resources being used to eradicate the disease in Bolivia should be concentrated in these endemic areas where convincing cattle owners of the need to control FMD is particularly difficult. It is also suggested that the eradication programme should coordinate its activities with neighbouring countries. PMID- 19284045 TI - [Design and implementation of a national serum bank for the surveillance of exotic animal diseases in the Republic of Cuba]. AB - A serum bank for the surveillance of exotic diseases was designed in accordance with the provisions of the Information and Epizootiological Surveillance System in the Republic of Cuba. Sera were collected from imported animals, from sentinel animals used for monitoring target areas at biological risk and from animals located in high animal-density areas. Methodologies were developed for the selection and characterisation of target areas at biological risk and sentinel animal points, the collection and storage of serum samples and the management of the national animal serum bank. After developing the methodologies, the serum bank was established throughout Cuba. The national animal serum bank operates using a quality management system based on the recommendations of the World Organisation for Animal Health and the International Organization for Standardization. PMID- 19284046 TI - Predicted trends in the supply and demand of veterinarians in Japan. AB - Currently in Japan, there are 32,000 active veterinarians, mainly engaged in small and large animal practice and public animal health and public health services. In the face of the notable increase in recent years in the proportion of female students enrolled in veterinary schools and in the number of households with companion animals, a model was developed to predict the supply and demand of veterinarians toward 2040 in Japan. Surveys were conducted on sampled households and veterinarians to estimate input variables used in the supply and demand model. From this data it is predicted that there might be somewhere between a shortage of 1,000 to an over-supply of 3,700 veterinarians engaged in small animal practice in 2040. This, however, will depend on possible changes in the number of visits made to veterinarians by small animal owners and the efficiency of practices in the future. The model also predicts that there will be a shortage of around 1,100 veterinarians in large animal practice in 2040. Considering the many assumptions made to estimate the input variables used in the model, the results of this study do not provide definitive conclusions, but provide a base for discussions on what will be needed in the veterinary profession in the future. PMID- 19284047 TI - [Evaluation of the safety of live attenuated vaccine viruses against infectious bursal disease (Gumboro disease) in conventional broiler chicks]. AB - The pathological and immunological effects of two infectious bursal disease (Gumboro disease) vaccines containing attenuated virus were evaluated under experimental conditions. A total of 200 conventional broiler chicks were divided into three batches, two of which were administered an experimental dose of Gumboro disease vaccine via the ocular route, at 14 days of age, using an 'intermediate' strain for one group and an 'intermediate plus' strain for the other. At different post-inoculation intervals, the chicks from the three batches were weighed and slaughtered and their bursa of Fabricius were removed to be weighed and subjected to histological examination. This allowed a lesion score and a ratio between the weight of the bursa of Fabricius and body weight to be calculated for each group of chicks. The immunosuppressive potential of the two vaccines was also estimated by evaluating the chicks' serological response to a Newcastle disease vaccine administered by eye-drop two weeks following vaccination against Gumboro disease. The results showed that the two vaccine strains were not of equal pathogenicity and that one was potentially immunosuppressive. The 'intermediate plus' strain proved to be more virulent than the 'intermediate' strain, as it caused greater atrophy and more severe lesions of the bursa of Fabricius. By exerting a more suppressive effect on the serological response of the chicks, the 'intermediate plus' vaccine also proved to be immunosuppressive. The three weeks of observation were not sufficient to ascertain whether the lesions caused by the two vaccines were temporary or whether the immunosuppressive state induced by the 'intermediate plus' strain was reversible. PMID- 19284048 TI - Control charts for identifying systematic errors using control sera to detect antibody to Salmonella in an indirect ELISA. AB - This study evaluated the preparation of Shewhart's control charts using the concept of rational subgroups for monitoring the Salmonella antibody ELISA used for surveillance of Danish pig herds. Control charts were prepared for a buffer control sample, a negative serum sample and a positive serum sample. The quality control variables were the natural logarithm (In) of the uncalibrated optical density (OD) for the buffer control sample and the negative serum sample, and the calibrated OD (OD%) for the positive serum sample. Testing round (run) within laboratory robot was chosen as the subgroup, and separate control charts were prepared for five robots. The control limits were set at six times the standard deviation for In(OD) and three times the standard deviation for OD%. Evaluation based on a number of sensitising rules for control charts produced from historical data showed that use of the control charts could reveal systematic analytical errors. PMID- 19284049 TI - A proficiency testing method for detecting antibodies against Brucella abortus in quantitative and qualitative serological tests. AB - A proficiency testing panel for detecting antibodies against Brucella abortus was developed and evaluated by both primary binding and conventional serological tests, using the guidelines of the World Organisation for Animal Health and the International Organization for Standardization Guide 43-1. All serological tests were judged satisfactory. Among the primary binding tests, the competitive enzyme linked immunosorbent assay (ELISA 2) and the indirect enzyme-linked immunosorbent assay (ELISA 1), with standard deviation indices (z-scores) of -0.06 and 0.10, respectively, performed best. Similarly, E(n) numbers (i.e. a way of comparing different measurements of performance) of 0 for both the competitive ELISA 2 and the indirect ELISA 1 indicated that these tests performed best in the initial round of proficiency testing. The conventional serological tests all passed the panel. Comparing data from both the quantitative and qualitative tests demonstrated that this proficiency testing scheme was fit for the purpose for which it was designed. PMID- 19284050 TI - Addressing the problems of using the polymerase chain reaction technique as a stand-alone test for detecting pathogens in aquatic animals. AB - The polymerase chain reaction (PCR) test is increasingly being used as the diagnostic tool of choice and with pathogens of aquatic animals, particularly molluscs and crustaceans, a PCR test is often the only laboratory diagnostic test available. When a diagnostic decision is required in the absence of clinical disease (e.g. health certification or a survey for disease freedom) limitations with the PCR technique may become apparent. A review of validated PCR tests demonstrates that most are imperfect assays with diagnostic sensitivities and/or specificities less than 100%. False positive and false negative results will thus occur and large numbers of samples need to be tested to achieve statistically significant results. The results from a survey for disease freedom from white spot syndrome virus in Australia are used to illustrate the problem and to assess various solutions. The use of pooled samples or the use of two independent PCR tests can substantially reduce the number of tests required and the associated costs of such a survey. PMID- 19284051 TI - Foot and mouth disease in the Lao People's Democratic Republic: I. A review of recent outbreaks and lessons from control programmes. AB - Foot and mouth disease (FMD) causes sporadic disease outbreaks in the Lao People's Democratic Republic (Lao PDR). As the Lao PDR is a major thoroughfare for transboundary animal movements, regular FMD outbreaks occur, causing economic hardship for farmers and their families. In this review of the recent history of FMD in the Lao PDR between 1997 and 2006, the authors examine the virological and epidemiological aspects of the disease and appropriate control measures, including the distribution of outbreaks, causative serotypes and the molecular epidemiology of the viruses, as well as large-scale vaccination programmes. The dominant serotype, type O, was reported every year from 1998 to 2005. The majority of outbreaks occurred in Vientiane Capital (n = 42; 28%) and the highest number of outbreaks were reported in cattle (n = 94; 61%); followed by buffalo (n = 41; 27%) and pigs (n = 18; 12%). All type A outbreaks occurred in cattle. Type Asia 1 outbreaks were reported in the central provinces around Vientiane Capital between 1996 and 1998. PMID- 19284052 TI - Foot and mouth disease in the Lao People's Democratic Republic: II. Seroprevalence estimates, using structured surveillance and surveys of abattoirs. AB - An examination of the seroprevalence of foot and mouth disease (FMD) virus was conducted in the Lao People's Democratic Republic (Lao PDR) from 1996 to 2005, using structured surveillance and abattoir-based studies. Under structured surveillance, seropositivity ranged from 65.7% (Vientiane Capital, 1996) to 3% (Houaphan, 2005) for cattle and buffalo; and from 2.8% (Vientiane Capital, 1998) to 0% in separate studies of pigs. In each study, species composition was significantly associated with seroprevalence rates. For abattoir surveys, the majority of samples (60.5%) came from Vientiane Capital (33.0%), Savannakhet (14.0%) and Champasak (13.5%) provinces. The overall proportion of animals testing positive for the presence of antibodies against the FMD virus was 18.7% (ranging from 50.8% in Vientiane Province to 1% in Phongsali). Generally, antibodies against serotype O were the most prevalent. Cattle and buffalo that tested as seropositive were significantly older than the seronegative animals (p < 0.00005). The overall proportional seropositivity was significantly different for different species, as was the case with the antibodies against serotypes O, A and Asia 1. Some 22% of cattle, 55% of buffalo and 23% of pigs demonstrated seropositivity but this varied significantly between provinces. PMID- 19284053 TI - Seroepidemiology of peste des petits ruminants in sheep and goats of southern peninsular India. AB - This paper presents the results of a seroepidemiological study carried out between July 2006 and March 2007 to detect the presence of antibodies to peste des petits ruminants (PPR) virus in randomly collected serum samples from sheep and goats in southern peninsular India. The authors used a competitive enzyme linked immunosorbent assay with a monoclonal antibody developed against a neutralising epitope of the haemagglutinin (HA) protein of the virus. A total of 1,492 sheep sera and 2,068 goat sera collected from the six southern Indian states were screened. It was determined that 41.35% of the sheep sera and 34.91% of the goat sera were positive for the presence of antibody. The study indicated an extensive endemicity of the disease in these states, which is attributed to the agro-climatic conditions and the migration of livestock. PMID- 19284054 TI - The epidemiology of peste des petits ruminants in Pakistan and possible control policies. AB - Peste des petits ruminants made its first clinical appearance in Pakistan during an epidemic in Punjab Province in 1991, but its level of activity in subsequent years was never systematically recorded. In 2003, 2004 and 2005 teams trained in participatory disease surveillance methods visited a large number of villages throughout Pakistan to record the prevalence and impact of key livestock diseases. Peste des petits ruminants has emerged from their study as a common and economically damaging disease of small ruminants within the country, appearing to be endemic in the north of Punjab and epidemic in Azad Jammu and Kashmir, and Sindh during the study period. It should be possible to reduce the incidence level using a combination of zoosanitary measures and the intensive use of vaccine. PMID- 19284055 TI - The epidemiology of peste des petits ruminants in Pakistan. AB - Peste des petits ruminants (PPR) is an acute and highly contagious viral disease of small ruminants, which is newly emerging in Pakistan. Information provided by participatory disease surveillance teams was used to develop a database for PPR outbreaks in Pakistan. Twenty-four villages were selected throughout the country and field investigations were conducted in each village to study the dynamics of the disease. In each area, flocks with a history of PPR were identified and serological sampling was conducted. Some 1,463 small ruminants (sheep and goats) were sampled and 1,096 tested positive for the presence of antibodies against PPR. These results clearly indicate that PPR is prevalent throughout Pakistan. PMID- 19284056 TI - Leptospira infection in animals and humans: a potential public health risk in India. AB - Leptospirosis is recognised as one of the commonest zoonotic infections in the world. In India, where animals provide the draught power for agriculture, which is the main profession of the population, the incidence of leptospirosis among animals and humans is high. In this paper, the isolation of pathogenic leptospires from human and animal hosts from several parts of India is reported. Because there are only limited facilities for serotyping within the country, most of the isolates were typed to the serogroup level only. In addition, the potential of leptospirosis to be a serious public health problem in India is discussed. PMID- 19284057 TI - Equine herpesvirus 1: characterisation of the first strain isolated in Colombia. AB - This paper describes the isolation and characterisation of equine herpesvirus 1 (EHV-1) in Colombia. The virus was isolated from a nasal swab and an aborted foetus of a pregnant mare imported from Argentina, with clinical signs of rhinopneumonitis. The new strain was characterised through culture and morphological, serological and immunocytochemical studies. Polymerase chain reaction and DNA restriction maps revealed an EHV-1 1P genome. This is the first report on the isolation and characterisation of EHV-1 in Colombia. PMID- 19284058 TI - The epidemiology of sheep pox in Greece from 1987 to 2007. AB - The authors reviewthe epidemiology of sheep pox outbreaks in Greece between 1987 and 2007. It is believed that sheep pox is introduced into Greece principally from neighbouring countries to the east, and is associated with the movements of infected sheep flocks close to the border and contacts between humans and animals. Disease foci have appeared in several central and north-eastern areas of the country. Between 1982 and 1986, Greece remained free of sheep pox but, in 1987, the disease appeared on the island of Lesvos and, in 1988, outbreaks were seen in the prefecture of Evros. In 1994, a further outbreak occurred in Evros. Over the next four years, more outbreaks occurred in Evros and Thessaloniki (1995); Larissa, Xanthi, Rhodopi, Kavala, Magnissia, Evros and the island of Lesvos (1996); Kavala, Magnissia, Halkidiki, Evros and Rhodopi (1997). In 1998, there were fewer cases of sheep pox, with outbreaks only in the prefecture of Evros. Two years later, a further outbreak was reported in Evros (2000), while the most recent outbreak occurred on the island of Lesvos in January 2007. PMID- 19284059 TI - Bluetongue infection rate in mithun (Bos frontalis) in the north-eastern upland region of India. AB - Studies conducted on 106 mithun at the National Research Centre on Mithun and 66 free-ranging mithun in Nagaland, India, revealed an infection rate with bluetongue virus of 86%, using a commercially available competitive enzyme-linked immunosorbent assay. Animals were grouped according to their age: 36 of 1 to 2 years of age, 50 of 2 to 4 years of age and 86 aged 4 years and over. The highest infection rate (98%) was found in mithun > 4 years old and the lowest (58%) in those 1 to 2 years old. No statistically significant difference was observed between infection rates of males (89%) and females (85%). The infection rate was higher (95%) in free-ranging mithun than in mithun kept under a semi-intensive system (80%). This is the first report of serological evidence of antibodies to bluetongue virus in mithun. The possible role of vectors in the epidemiology of bluetongue virus infection in mithun is discussed briefly. PMID- 19284060 TI - Status of bovine tuberculosis in Addis Ababa dairy farms. AB - The study was conducted to determine the status of bovine tuberculosis in Addis Ababa, Ethiopia, by a comparative intradermal tuberculin test of 1,869 animals in 106 farms. Epidemiological information was also collected, taking into account factors chosen for their epidemiological significance and local livestock husbandry characteristics. In addition, milk samples were collected from tuberculin reactors for mycobacterial isolation and characterisation. Chi-square statistic, simple regression and multiple stepwise logistic regression were used to analyse the data. Of the 106 farms examined, 46 (95% confidence interval [CI]: 33.8% to 53.4%) contained comparative skin test reactors. Of the 1,869 animals, 443 (95% CI: 21.8% to 25.7%) were comparative skin test reactors. Furthermore, about 8.5% of tuberculin sensitive cows (12 of a sample of 141) secreted acid fast bacteria in their milk. The microbes are described in more detail in the paper. Factors identified as possibly increasing the risk of bovine tuberculosis in Addis Ababa were herd size (large herd), farming (housing) condition (poor), and age (older animals). Similarly, as body condition scores improved from poor to medium and then to good, the likelihood of positive results significantly decreased (OR = 0.54; p < 0.01). Other factors including breed, sex, and physiological status of animals did not seem to significantly contribute to tuberculin sensitivity. The finding that large-size and intensively (often poorly) managed herds were at greater risk of bovine tuberculosis suggests that the significance of bovine tuberculosis is increasing in Addis Ababa parallel to an increasing dairy operation. If measures are not taken promptly, the impact on the economy and public health could be enormous. PMID- 19284061 TI - The gender paradox. PMID- 19284062 TI - Observational studies of drug-eluting stents--some are more equal than others. PMID- 19284063 TI - Meeting report ESC forum on drug eluting stents, European Heart House, Nice, 27 28 September 2007. PMID- 19284064 TI - Effect of percutaneous mitral repair with the MitraClip device on mitral valve area and gradient. AB - AIMS: Percutaneous repair of mitral regurgitation (MR) by leaflet apposition using a clip deployed via transseptal catheterisation is undergoing evaluation. METHODS AND RESULTS: In order to detect the potential for clinically significant left ventricular inflow obstruction after percutaneous repair, we measured mitral valve area (MVA) and mean transmitral gradient (MVG) echocardiographically in 96 patients implanted with a clip followed for up to 24 months. By planimetry, the mean MVA decreased from 6.0 +/- 1.3 cm2 to 3.6 +/- 1.2 cm2 (p < 0.05) (range 1.9 to 7.6 cm2) after clip placement, and remained unchanged after 24 months of follow-up (3.5 +/- 0.8 cm2). The mean MVG increased after clip placement from 1.7 +/- 0.9 mmHg to 4.1 +/- 2.2 mmHg (p < 0.05), and did not increase further to 24 months (3.8 +/- 1.9 mmHg). There were no differences in MVA or MVG between patients who received 1-clip (69%) and those receiving 2-clips (31%). Patients with functional MR (23%) had a slightly smaller MVA, both at baseline and after clip placement, but did not differ from degenerative MR patients at later follow up. After 2 years of follow-up, no patient required surgery for LV inflow obstruction. CONCLUSIONS: Mitral repair with the MitraClip device for MR decreases MVA without significant mitral obstruction. After 2 years of follow-up, no patient required surgery for LV inflow obstruction, and these results were not influenced by the use of more than 1 clip or the aetiology of MR. PMID- 19284065 TI - Assessment of the absorption process following bioabsorbable everolimus-eluting stent implantation: temporal changes in strain values and tissue composition using intravascular ultrasound radiofrequency data analysis. A substudy of the ABSORB clinical trial. AB - AIMS: The main objective was to use IVUS-backscatter radiofrequency (IVUS-RF) to assess the degradation of a bioabsorbable stent by measuring serial changes in dense calcium (DC) and necrotic core (NC) as assessed by intravascular ultrasound Virtual Histology (IVUS-VH) and in the strain as assessed by palpography. METHODS AND RESULTS: In the ABSORB trial, 27 patients treated with a single bioabsorbable everolimus-eluting stent (BVS, Abbott Vascular, Santa Clara, CA, USA) were all imaged with IVUS-RF post-stenting and at 6-month follow-up, and 13 and 12 patients were also investigated pre-stenting with IVUS-VH and palpography respectively. From pre- to post-stenting, with VH (n = 13), there was an increase in mean "DC" (9.8 vs. 25.4%, p = 0.0002) and "NC" (15.5 vs. 30.5%, p = 0.0002). In palpography (n = 12), the mean number of frames with Rotterdam Classification (ROC) III/IV per cm decreased from 1.22 +/- 1.91 to 0.12 +/- 0.31 (p = 0.0781) and the mean cumulative strain values (all frames with ROC I-IV scores) changed from 0.50 +/- 0.27 to 0.20 +/- 0.10% (p = 0.0034). Comparing post-stenting with follow-up (n = 27), VH showed a decrease in "DC" (29.7% vs. 21.1%, p = 0.0001). "NC" also decreased (26.9 vs. 21.5%, p = 0.0027). For palpography (n = 25 patients), an increase in the mean number of frames with ROC III/IV per cm was observed from 0.09 +/- 0.26 to 0.22 +/- 0.36 (p = -0.1563) while the mean cumulative strain values (all frames with ROC I-IV scores) changed from 0.15 +/- 0.10 to 0.26 +/- 0.12% (p < 0.0001). CONCLUSIONS: IVUS-VH changes at 6 months suggest alteration of the BVS with reduction of RF backscattering by polymeric struts. Strained plaques on the palpograms were almost abolished following stent implantation. However, strain values reappeared within 6 months suggesting an increase in endoluminal deformability of the stented vessel. PMID- 19284066 TI - Twelve month clinical and angiographic outcome after stenting of unprotected left main coronary artery stenosis with paclitaxel-eluting stents--results of the multicentre FRIEND registry. AB - AIMS: To evaluate the angiographic and clinical outcome of patients undergoing paclitaxel-eluting stent (PES) implantation for unprotected left main coronary artery (ULMCA) stenosis in a multicentre, prospective registry. The overall event rate for PCI of ULMCA disease remains higher than in on-label use making additional outcome data and risk-stratification tools for the ULMCA population desirable. METHODS AND RESULTS: A prospective registry included all patients with a significant (> 50%) stenosis in ULMCA disease. In 151 of these patients the target lesion involved the distal bifurcation in 100 patients (66%), which was treated by predominantly using a "provisional T stenting" strategy. In distal ULMCA disease group, 72% had only one stent implantation while 28% had multiple (either 2 or 3) stents implanted. At a median follow-up of 472 +/- 75 days, cardiac death occurred in 3 patients (2%) and major adverse cardiac and cerebrovascular events (MACCE) in 16 patients (10.6%). CONCLUSIONS: In the drug eluting stent era, paclitaxel eluting stent implantation of ULMCA stenosis provided excellent immediate and mid-term results in this selected population, suggesting that it may be considered as a safe and effective alternative to CABG for selected patients with ULMCA who are treated in institutions performing large numbers of PCI procedures. PMID- 19284067 TI - Long-term follow-up of percutaneous coronary intervention of unprotected left main lesions with drug eluting stents: predictors of clinical outcome. AB - AIMS: To evaluate the long-term follow-up of drug-eluting stents (DES) in the treatment of unprotected left main coronary artery (ULMCA). METHODS AND RESULTS: One hundred and forty-eight patients (mean age 71 +/- 10 years) with ULMCA stenoses underwent percutaneous coronary intervention (PCI) with DES. Mean ejection fraction (EF) was 63 +/- 13% and distal ULMCA was involved in 63.5% of cases. In-hospital outcome showed one intra-procedural death, no stent thrombosis and 2% non Q-wave myocardial infarction (MI). Clinical follow-up was available in all patients (874 +/- 382 days): 10.1% of them had died, 8.8% had target lesion revascularisation (TLR) and 4.1% experienced MI. Major adverse cardiac events (MACE) occurred in 20.3%. Mortality predictors were EF < or = 55% (OR 3.6, 95% C.I. 1.3-10.1, p = 0.016) and EuroSCORE > or = 6 (OR 3.9, 95%-CI 1.1-14.1, p = 0.037). TLR predictors were distal lesion (OR 8.5, 95%-CI 1.1-15, p = 0.041) and age < 64 years (OR 3.1, 95%-CI 1-9, p = 0.042). MACE predictor was EF < or = 55% (OR 2.4, 95%-CI 1.1-5.2, p = 0.027). CONCLUSIONS: ULMCA stenting with DES is safe, with favourable in-hospital outcome. Long-term results are acceptable with a mortality rate of 10%, a TLR rate of 9%, and a MACE rate of 20%. Low EF and high EuroSCORE predict mortality, while younger age and distal lesions predict TLR. Low EF also predicts MACE. PMID- 19284068 TI - The relative safety and efficacy of bare-metal and drug-eluting stents in low and high-risk patient subsets. An epidemiological analysis of three sequential cohorts of consecutive all comers (n = 6129). AB - AIMS: Sirolimus- and paclitaxel- eluting stents (SES and PES respectively) have been shown to produce a sustained reduction in restenosis and repeat revascularisations as compared to bare-metal stents (BMS) up to four years. There is still limited data about the long-term safety and efficacy of DES in high-risk subgroups. METHODS AND RESULTS: A total of 6,129 consecutive patients were treated during three sequential periods with BMS (n = 2,428; January, 2000 to April, 2002), SES (n = 866; April 2002 to February 2003) or PES (n = 2,835; February 2003 to December 2005). A stratified analysis (including age, gender, diabetes, clinical presentation, treated vessel, multivessel disease, AHA lesion class, bifurcation, in-stent restenosis, average stent diameter < or = 2.5 mm and total stented length < or = 30 mm) was performed to evaluate possible heterogeneities in treatment effect. At four years, all-cause mortality was identical between the drug-eluting stent (DES) and BMS cohorts (13.5% vs. 13.4%, respectively; Adjusted HR 1.10, 95% CI 0.90 - 1.34) without evidence of heterogeneity in the high-risk patient subsets. Both DES significantly reduced the risk for target vessel revascularisation (TVR) as compared to BMS (TVR: 11.9% vs. 15.7% respectively; Adjusted HR 0.69, 95% CI 0.58 - 0.82) along with a reduced risk for post-operative MI (adjusted HR 0.75, 95% CI 0.57 - 0.98), but counterbalanced by a non-significantly higher risk for stent thrombosis (3.1% vs. 1.6%; adjusted HR 1.26, 95% CI 0.82 - 1.95). DES failed to show superiority to BMS in patients with acute myocardial infarction (TVR 10.5% vs. 9.2% respectively; Adjusted HR 1.26, 95% CI 0.82 - 1.93). CONCLUSIONS: In a real world patient population, after four years, the overall use of DES was associated with similar all-cause mortality rates and a significantly reduced risk for post operative MI and TVR as compared to BMS. PMID- 19284069 TI - Dual quantitative coronary angiography: a novel approach to quantify intracoronary thrombotic burden. AB - AIMS: To validate and test in vivo a new modality of quantitative coronary angiography (QCA), dual QCA (D-QCA), developed to quantify intracoronary thrombotic burden (ITB). METHODS AND RESULTS: Calculation of ITB with D-QCA is based on the discrepancy of luminal areas assessed with edge detection (ED) and video-densitometry (VD), measured with Cardiovascular Angiography Analysis System II. Experimental validation was first performed in phantoms with known obstructive volumes. In vivo assessment of thrombotic burden changes was performed in angiograms from 19 patients with large ITB, obtained before and after antithrombotic treatment, and compared with semi-quantitative assessment (TIMI thrombus grade (TTG)). A good correlation between D-QCA and true occlusive volumes was found (y = 9.21+0.99x, r = 0.996). Intra- and inter-observer variability was 2.77 +/- 10.97 mm3 (p = 0.50) and -1.28 +/- 6.99 mm3 (p = 0.62) respectively. In vivo, D-QCA demonstrated a significant reduction in ITB resulting from treatment (137.22 +/- 120.13 mm3 before and 104.72 +/- 99.19 mm3 after treatment, p = 0.001). Overall, TTG also decreased (3.63 +/- 0.68 before and 3.11 +/- 1.20 after, p = 0.008), but in those nine (47%) patients in which remained unchanged D-QCA detected a reduction in ITB (pre 148.17 +/- 154.03 mm3, post 112.86 +/- 117.82 mm3, p = 0.05). CONCLUSIONS: D-QCA appears as a useful approach to quantify IC thrombus volume, being more sensitive than TTG in assessing changes in ITB resulting from treatment strategies. PMID- 19284070 TI - Initial evidence for the return of coronary vasoreactivity following the absorption of bioabsorbable magnesium alloy coronary stents. AB - AIMS: To investigate the endothelium independent coronary smooth muscle vasomotor function four months after implantation of magnesium alloy absorbable metal stents (AMS) as part of the Progress-AMS clinical trial (n = 5), compared with a control group of patients implanted with permanent metal stents (PMS) (n = 10) undergoing follow-up angiography, but who were free from angiographic restenosis. METHODS AND RESULTS: Quantitative coronary angiogram (QCA) using an automated edge detection system was performed before and after the administration of 2 mg intracoronary isosorbide dinitrate (ISDN). The vessel diameter was measured at 0.2 mm intervals throughout the stented segments and a 1 cm proximal reference segment. The cross sectional area (CSA) was calculated before and after the ISDN, averaged and the percentage change measured. Reference segments demonstrated preserved vasomotor function in all cases: +13.28% (AMS) versus +17.15% (PMS), p = 0.39. The mean percentage increase in CSA for the stented segment was +6.78% for the AMS versus -1.30% for PMS, p = 0.003. CONCLUSION: These data demonstrate that four months after AMS implantation vasomotor function in reference segments is no different to that observed with PMS. However in contrast to PMS, within the AMS-stented segments there is demonstrable vasodilatation. These observations may have important implications for future stent design. PMID- 19284071 TI - Predictors for very late stent thrombosis after drug-eluting stent implantation in diabetic patients. AB - AIMS: Despite encouraging results with drug-eluting stents (DES) reported in diabetic patients, the long-term safety is unknown because of very late stent thrombosis (VLST). We investigated the incidence, risk factors and clinical manifestations of VLST in diabetic patients treated with DES, during long-term clinical follow-up. METHODS AND RESULTS: A total of 610 consecutive diabetic patients underwent PCI with DES. Dual antiplatelet treatment (APLT) for 12 months received 93%, more than 12 months 72% and statin treatment 93% of patients. Clinical follow-up of at least 12 months post DES implantation was obtained in 597/610 (98%) patients. The incidence of VLST was 1.8%, and 1.7% of patients developed stent thrombosis (ST) up to 12 months. All patients with VLST presented with sudden cardiac death and 82% were on dual APLT at the time of the event. In a multivariate model the only predictor for VLST (HR: 20.58, 95% CI 5.17-81.90, p < 0.001) and overall ST (HR: 4.38, 95% CI 1.73-11.10, p = 0.002) was ejection fraction < 40%. CONCLUSIONS: The incidence of ST in diabetic patients undergoing PCI with DES and receiving dual APLT is low at long-term clinical follow-up. The only predictor for VLST and overall ST was depressed left ventricular systolic function. PMID- 19284072 TI - Effect of gender differences on early and mid-term clinical outcome after percutaneous or surgical coronary revascularisation in patients with multivessel coronary artery disease: insights from ARTS I and ARTS II. AB - AIMS: The aim of the current study was to compare the short and mid-term outcome between males and females treated with percutaneous coronary intervention (PCI) with bare metal stent implantation or coronary artery bypass graft (CABG) surgery and drug-eluting stent implantation in the Arterial Revascularisation Therapies Study I and II (ARTS I and II). METHODS AND RESULTS: The patients included in ARTS I were randomised to PCI with bare metal stents or to CABG. The patients enrolled in ARTS II were treated with Cypher stent implantation. All patients were scheduled for clinical follow-up at one, six and twelve months, and after three and five years. Major adverse cardiac and cerebrovascular events (MACCE) included death, cerebrovascular accident (CVA), myocardial infarction (MI), repeat target vessel PCI (RPCI) and CABG. At one and three-year follow-up in ARTS II, both the female and male patients had an incidence of MACCE similar to ARTS I CABG. When comparing the female and male population of ARTS II, there were no differences between the two genders in terms of in-hospital outcome. At one year and three years there were no gender specific differences in the incidence of MACCE. CONCLUSIONS: Female and male patients in ARTS II had significantly lower MACCE rates compared with ARTS I-PCI, but similar to that of ARTS I-CABG. In ARTS II, MACCE free survival was similar for the two genders at three years follow-up. PMID- 19284073 TI - Clinical investigation into the observation that silicon carbide coating on cobalt chromium stents leads to early differentiating functional endothelial layer, increased safety and DES-like recurrent stenosis rates: results of the PRO Heal Registry (PRO-Kinetic enhancing rapid in-stent endothelialisation). AB - AIMS: Recurrent stenosis and stent thrombosis are still major concerns after drug eluting stent placement which inhibits not only the restenostic process but endothelialisation as well. In contrast, through accelerating rapid endothelialisation and development of an earlier functional endothelial layer, passive coatings have shown encouraging results. The objective of the present study was to investigate the clinical outcome and rate of recurrent stenosis of silicon carbide passive coated cobalt chromium stents (PROKinetic Coronary Stent with PROBIO coating, Biotronik AG, Switzerland) on restenosis after percutaneous coronary intervention. METHODS AND RESULTS: Percutaneous coronary stent deployment was carried out in 161 lesions in 145 consecutive patients. The primary combined endpoint was the rate of target-lesion revascularisation (TLR) and late lumen loss; the secondary endpoints were the procedural success and the major adverse cardiac events at 6-months follow-up. Out of 145 patients, 141 were successfully amenable to a silicon carbide coated stent (PRO-Kinetic, Biotronik AG, Switzerland) implantation (97.2% procedural success). At follow-up, the late loss was 0.75 +/- 0.71 mm. (in-stent) respectively 0.79 +/- 0.72 mm (in-segment), TLR was 4.9% and MACE was 5.6%. CONCLUSIONS: By augmenting rapid endothelialisation and development of an earlier functional endothelial layer, silicon carbide (PROBIO) as a passive coating on cobalt chromium stents has shown encouraging results relative to success rates, clinical outcome, TLR and late loss in a cohort of patients with extended coronary artery disease. PMID- 19284074 TI - Integration of 3D reconstruction in the SELection criteria for Excessive Crossing Times for Magnetically Supported Percutaneous Coronary Intervention. SELECT-MP. AB - AIMS: To develop a clinical prediction rule based on 3D reconstruction of coronary arteries that would prospectively identify lesions that are difficult to cross and could benefit from magnetic navigation. METHODS AND RESULTS: The coronary anatomy of a cohort of 120 lesions that had undergone percutaneous coronary intervention (derivation set) was analysed using 3D reconstruction for vessel and lesion characteristics. The crossing time was the total clock time to reach a satisfactory distal position after leaving the guiding catheter. Multivariable logistic regression and linear shrinkage with bootstrapping were used to develop a clinical prediction rule that dichotomised cases into easy or difficult (prolonged crossing time). A value of 6 was the best cut-off value. This clinical prediction rule was applied to a second independent cohort of patients (validation set) where crossing time was measured. The bootstrapped c statistic of the model was 0.82 indicating excellent discrimination. CONCLUSIONS: 3D reconstruction helped to develop a simple, accurate clinical prediction rule to identify difficult cases for conventional wires and in whom magnetic navigation may be preferable. PMID- 19284075 TI - Magnetically navigated percutaneous coronary intervention in distal and/or complex lesions may improve procedural outcome and material consumption. AB - AIMS: Comparison of magnetic guidewire navigation in percutaneous coronary intervention (magnetic PCI) across distal and/or complex lesions versus conventional navigation (conventional PCI). METHODS AND RESULTS: Forty-seven consecutive patients (age 61 +/- 10 yr) undergoing elective single vessel magnetic PCI for distal and/or complex lesions were matched by age and lesion location with 45 patients undergoing conventional PCI (age 63 +/- 10 yr). Technical success rate was defined as an intraluminal wire position distal to the stenosis. Procedural outcome and costs were evaluated. Baseline demographics and angiographic characteristics of the two groups were similar. The technical success rate did not differ between magnetic and conventional PCI (95.7 vs 97.8%; p = 1.00). Significantly shorter procedural and fluoroscopy time were observed for magnetic compared to conventional PCI (29.9 +/- 17.6 vs 41.1 +/- 21 min, p = 0.007; 7.5 +/- 7.3 vs 16.1 +/- 22.4 min, p = 0.02 respectively). Less contrast was used in the magnetic PCI group (58 ml/patient; P = 0.02). These advantages resulted in a mean estimated saving of 1400 euro per patient (P < 0.001). Advantages of procedural outcome were even more pronounced in the ACC/AHA lesion class C subgroup. CONCLUSIONS: Magnetic compared to conventional PCI is an attractive novel technique that proved to be feasible and safe and might be faster in distal and especially complex lesions. PMID- 19284076 TI - Efficacy and safety of enoxaparin in combination with and without GP IIb/IIIa inhibitors in unselected patients with ST segment elevation myocardial infarction treated with primary percutaneous coronary intervention. AB - AIMS: We sought to determine the efficacy of enoxaparin in unselected patients with STEMI treated with primary percutaneous coronary intervention in clinical practice. METHODS AND RESULTS: In a retrospective analysis of the prospective MITRA-plus registry we compared the outcomes of patients with primary PCI and either enoxaparin or unfractionated heparin. A total of 2,655 patients with STEMI < 12 hours were included in this analysis, 374 (14%) were treated with enoxaparin and 2,281 (86%) with unfractionated heparin. In the univariate analysis enoxaparin reduced mortality (1.6% versus 6.0%, < 0.001), fewer non-fatal reinfarctions (1.9% versus 3.8%, p = 0.05) and no significant difference in major bleeding (5.6% versus 7.2%, p = 0.2) was observed. In the multivariable propensity score analysis enoxaparin was associated with a reduction in the combined endpoint of death and non-fatal reinfarction (odds ratio 0.42; 95% CI 0.2-0.8). This advantage was observed both in subgroups without (odds ratio 0.33 95% CI 0.1-0.8) and with GP IIb/IIIa inhibitors (odds ratio 0.44, 95% CI 0.2 1.0). CONCLUSIONS: Our data suggest that in unselected patients with STEMI treated with primary PCI enoxaparin compared to unfractionated heparin reduces the combined endpoint of in-hospital death and reinfarction and does not increase severe bleeding complications. PMID- 19284077 TI - Current and future developments in intracoronary optical coherence tomography imaging. AB - Optical coherence tomography (OCT) has become a key intracoronary imaging modality able to traverse some of the limitations of angiography and intravascular ultrasound. In vivo imaging with high resolution (around 15 micrometres) has given unique insights into not only atherosclerotic plaque, but also to the understanding of tissue responses underlying stent implantation. Novel developments with faster OCT pullback speeds will further simplify the procedural requirements and eventually eliminate the need for proximal vessel balloon occlusion during image acquisition. This report explores the current and future developments in OCT technology that will see this unique imaging modality become a key player in both the clinical and research arena for the interventional cardiologist. PMID- 19284078 TI - In vivo evaluation of a biolimus eluting nickel titanium self expanding stent with overlapping balloon expandable drug eluting and bare metal stents in a porcine coronary model. AB - AIMS: Long lesions and complex vessel anatomy frequently require the use of overlapping stents to treat a lesion. The purpose of this study was to evaluate the long-term effects of overlapping the Axxess Biolimus A9 eluting stent (BES) with two of the most commonly used, commercially available drug eluting stents. These stents were compared to BxVelocity bare metal (BMS) stents in a porcine coronary stent-injury model. METHODS AND RESULTS: Nineteen juvenile farm swine, 25-35 kg in weight, 3-6 months in age were utilised. Each animal received an Axxess stent to their coronary artery as permitted by the individual animal's anatomy. A second stent, either a Cypher, sirolimus eluting stent (SES) or, a Taxus, paclitaxel eluting stent (PES), or a BxVelocity bare metal stent (BMS) were implanted in an overlapped fashion. The animals were then followed for either 28 or 180 days as specified by a randomisation scheme. At the end of each follow-up period, they were euthenised, and the vessels containing the overlapping stents were harvested, processed into histological sections, and analysed. Compared to bare metal stents, overlapped segments using DES exhibited delayed vascular healing compared to both the proximal and distal non-overlap sites at each of the follow-up time point. Overall, in the non-overlap stent segments, SES induced significantly more inflammation and neointimal hyperplasia compared to PES and BMS. CONCLUSIONS: In this study of BMS and two different types of DES overlapped with the Axxess Biolimus A9 eluting stent, we found that while there was a delay in the degree of vascular healing with DES compared to BMS, the specific type of DES that was overlapped with BES did not affect the behaviour of the overlap zone in terms of most of the histomorphometric measures at 28 or 180 days. This was true whether the stent was drug eluting or bare metal. More inflammation with delayed healing was seen in the SES compared to PES and BMS. PMID- 19284079 TI - Coronary steal with unstable angina secondary to a coronary artery fistula. AB - BACKGROUND: A 62-year old man with progressive deterioration of stable angina presented with an unstable episode and was referred for diagnostic coronary angiography. INVESTIGATIONS: Physical examination, electrocardiography, myocardial perfusion scan, coronary angiography, cardiac CT. DIAGNOSIS: Left coronary main stem to pulmonary artery fistula. MANAGEMENT: Coronary angiography, percutaneous coronary intervention, coil embolisation. PMID- 19284080 TI - Lipolytic enzymes, metablic syndrome and atherosclerosis (revisiting and revising a 40 years old study). AB - More than 40 years ago, our laboratory reported that post-heparin lipolytic activity was decreased in patients with severe atherosclerotic disease, while values recorded in obese and hyperlipidemic subjects without clinically detectable atherosclerotic lesions did not significantly differ from normal weight normolipidemic controls. Because in 1967 data on pathophysiology of lipolytic enzymes were rather scarce, and mainly because our information facilities were limited in those years, we had difficulties in interpreting these results, and the study was to some extent awkwardly approached, as the investigated subjects were not considered according to their gender, body fat patterning and type of hyperlipoproteinemia, and the lipolytic activities of lipoprotein lipase and hepatic lipase had not been selectively assessed. Reviewing recent data in the literature it was noted that pre-heparin lipoprotein lipase mass assessed by ELISA was indeed significantly lower in insulin resistant coronary patients than in patients with no lesions, and correlated negatively with the severity of atherosclerotic lesions. Noteworthy hypoadiponectinemia, a hallmark of insulin resistance, was associated with decreased lipoprotein lipase and increased hepatic lipase activities. Clustering of increased plasma VLDL triglyceride, cholesteryl-ester transfer protein and hepatic lipase would remodel HDL and LDL particles, generating an atherogenic lipoprotein profile. In opposition to atherogenic dyslipidemia related to an enhanced hepatic secretion of VLDL, cases with important hypertriglyceridemia subsequent to deficient lipolytic clearance are at a rather low risk for coronary artery disease. It may therefore be suggested that the decreased lipoprotein lipase noted in atherosclerotic patients is not a major pathogenic link, being rather related to the inflammatory component of the disease, its expression being reduced by proinflammatory cytokines. PMID- 19284081 TI - Impact of host insulin resistance on fibrosis and response to interferon in chronic hepatitis C. AB - Chronic hepatitis C and obesity inflict significant health and economic burdens on the world. Insulin resistance associated to obesity is the key factor in this disease, mainly because it has been related to fibrosis extension and non response to antiviral therapy. The mechanism underlying this correlation is not fully explained yet. Leptin, adiponectin, TNF alfa, the hepatic expression of suppressor of cytokine signalling 3 (SOCS3), and hyperinsulinemia have been considered as heavily influencing fibrosis extension and nonresponsiveness to the IFN-alpha. The adipokines increased hepatic expression of SOCS3, and hyperinsulinemia has been proposed as heavily influencing non-responsiveness to the IFN-alpha and fibrosis extension by maintaining the hepatic stellate cells activated phenotype in patients with chronic hepatitis C and insulin resistance related obesity. We shall review the mechanisms by which obesity-related insulin resistance may be associated with fibrosis extension and decreased efficacy of IFN-alpha based therapies in obese individuals with chronic hepatitis C and the therapeutic strategies that may increase the effectiveness of these therapies. Consequently, being aware of the role of obesity-related insulin resistance in chronic hepatitis C, it will be possible to elaborate new therapeutic strategies for chronic hepatitis C obese patients. Also, it will be possible to elaborate new criteria for the initiation of antiviral therapy by including parameters such as body weight, abdominal circumference, HOMA-IR, the dosage of adipokines in order to more accurately define the groups of patients with a higher risk of non responsiveness to antiviral therapy, thus lowering long-term costs. PMID- 19284082 TI - Bronchial asthma with psychogenic trigger. AB - The authors present several personal studies in accordance with other published data which contest the existence of purely psychogenic bronchial asthma, as it was considered 30-50 years ago. Although psychological stimuli could trigger the occurrence of attacks of bronchial asthma, they are not an exclusive etiological factor (Iamandescu, 1980, 1985). In the context of large groups of asthma with allergic or other etiology, forms of asthma that are triggered by an additional psychogenic factor could be identified. These are noted in 62.5% of hospitalized patients, most cases with a prolonged and more severe evolution of the disease, but in only 30% of light and moderately severe cases that are followed up in outpatient (Iamandescu, 1980, 1984). The percent of psychological trigger is maximum (86%) in corticodependent asthma patients (Iamandescu, 1996). Somatic and psychological background of asthma with psychogenic triggering is represented by the following: (1) an obvious relationship between psychological stimuli and the onset of asthma attacks, and a psychological terrain "endowed" with an increased vulnerability to stress. These patients present a diminished rate of cortisol response to stress compared with asthma patients without psychogenic triggering; (2) reversibility to broncho-dilator agents, in contrast with resistance to systemic corticoid therapy; (3) sensitivity to aspirin; (4) endocrine dysfunctions (high estrogen and thyroid hormones secretion); (5) multi-factorial triggering of the attacks (especially non-specific respiratory irritants, including physical-chemical factors and meteorological changes) (1985); (6) variable response to psychotherapy and psychotropic drugs, suggesting--in case of therapeutic failure--the predominance of the chronic inflammation determined by other etiologic agents, different from psychological stress. PMID- 19284083 TI - The involvement of genetic factors in chronic venous insufficiency. AB - Chronic venous insufficiency (CVI) represents an important medical and social problem with a significant impact on the quality of life. Although the mechanisms of occurrence and development of chronic venous insufficiency became better understood in the last decades, the contribution of genetic risk factors is not precisely established. Many factors are involved in the etiopathogenesis of CVI, such as age, sex, heredity, sedentary life style. The impact of genetic factors is variably estimated. Genes polymorphisms associated with hyperhomocysteinemia, coagulation abnormalities, genetic factors involved in venous thrombosis leading to secondary CVI, represent the genetic background of this complex disease. PMID- 19284084 TI - Oxygen, a paradoxical element? AB - Oxygen is an essential element for life on earth. No life may exist without oxygen. But in the last forty years, conclusive evidence demonstrated the double edge sword of this element. In certain conditions, oxygen may produce reactive species, even free radicals. More, the production of reactive oxygen species (ROS) takes place everywhere: in air, nature or inside human bodies. The paradox of oxygen atom is entirely due to its peculiar electronic structure. But life began on earth, only when nature found efficient weapons against ROS, these antioxidants, which all creatures are extensibly endowed with. The consequences of oxygen activation in human bodies are only partly known, in spite of extensive scientific research on theoretical, experimental and clinical domains. PMID- 19284085 TI - Lipoprotein (a) and lipid and non-lipid risk factors in coronaries risk assessment. AB - PURPOSE: Several studies showed that elevated plasma levels of lipoprotein(a) [Lp(a)] represent a predictor for cardiovascular risk. Based on already existing literature data, we aim to study the relationship between Lp(a), lipids and other cardiovascular risk factors in individuals with or without coronary heart disease. METHODS: We performed a cross-sectional transversal study on 208 patients (100 men and 108 women) aged between 37-75, with or without old myocardial infarction. In all the patients were evaluated the cardiovascular risk factors, the plasma level of the lipid fractions and Lp(a). The relationship between Lp(a) and the lipid and non-lipid risk factors were evaluated by the logistic regression method. RESULTS: The myocardial infarction group had higher values of plasma levels of Lp(a) (0.37 +/- 0.28 vs. 0.29 +/- 0.23 g/L, p < 0.05), and LDL-C (125.66 +/- 41.21 vs. 113.44 +/- 46.64 mg/dL, p < 0.05), than the group without coronary heart disease, as well as higher values of plasmatic TC/HDL-C ratio (4.31 +/- 1.55 vs. 4.08 +/- 1.29, p < 0.05), with significantly decreased plasmatic levels of HDL-C (45.88 +/- 12.04 vs. 53.22 +/- 23.12 mg/dL, p < 0.05). The association between the high Lp(a) plasma levels and the severity of coronary vessels number involved was significant. Multivariate analysis performed with adjustments for cardiovascular risk factors showed that the Lp(a), LDL-C and CT/HDL-C ratio levels are significant and independent predictive markers of coronary heart disease. CONCLUSION: The results show that the high Lp(a) plasma levels represent an independent cardiovascular risk factor, with superior risk prediction than the conventional lipid fractions. Our results confirm the Lp(a) as a marker for cardiovascular risk assessment in clinical practice. PMID- 19284086 TI - A new prediction score for myocardial infarction: MINF SCORE. AB - The present report aims to generate a simple and efficient non-invasive prediction system of myocardial infarction as well as evaluating of the new generated score. METHODS: 208 patients (both men and women) aged 37-75 admitted at Centre Hospitalier Coulommiers, France, were included in the study. Patients were divided into two groups, according to presence (104 patients, 63.7 +/- 9.2 years) and absence of myocardial infarction (104 patients, 58.1 +/- 12.4 years). RESULTS: Based on univariate and multivariate analyses the following parameters were selected to form the prediction model: age, lipoprotein (a), C-reactive protein, systolic blood pressure, non-HDL-Cholesterol. A new score was obtained and it was validated by the area under the ROC curve of 0.745 +/- 0.034 (95% CI = 0.681-0.804), p < 0.0001. For the newly generated score, at the cut-off point (> 0.3), we obtained the following accuracy parameters: sensibility=80%, specificity=62.5%, positive predictive values=65%, negative predictive values =78%. The mentioned parameters were superior to values of each component of the score individually analyzed. CONCLUSIONS: The newly generated system can become a useful tool in coronary risk evaluation, with direct application in clinical practice. PMID- 19284087 TI - Hypertensive acute pulmonary oedema as expression of diastolic heart failure. AB - In part of hypertensive acute pulmonary oedema patients we can determine normal ejection fraction (EF) after remission of symptomatology and pulmonary congestion. In most cases echocardiography is performed after the disappearance of pulmonary oedema (APO). METHODS: Echocardiography was performed in standard views. The EF was measured by area-length monoplane method in apical 4-chamber and 2-chamber view, the final figure being an arithmetical mean of the two values. Measurement of the EF was made during APO and immediately after disappearance of symptoms and pulmonary congestion. Color, pulsed and continued Doppler were used for quantification of mitral regurgitation in apical 4-chamber and 2-chamber view. RESULTS: The study included 61 patients with hypertensive APO, with systolic blood pressure (BP) >160 mmHg. From the total, 37 (60.65%) were men and 24 (39.34%) female and the mean age was 65.02 +/- 12.17 years. The systolic blood pressure (SBP) during APO was 196.97 +/- 18.89 mmHg and after treatment 132.38 +/- 11.78 with significant statistical power (p < 0.0001), with 95% confidence interval from -71.083 to -57.553. EF during APO (EF-APO) was 49.84 +/- 10.7 and after disappearance of symptomatology and pulmonary congestion (EF nonAPO) it was 48.9 +/- 8.47. The distribution of EF values per patients is illustrated in the figure below. The values of EF-APO are significantly correlated with EF-nonAPO values (p < 0.0001, r2 = 0.9093, alpha=0.05). The linear regression of EF values during and after APO is significant (r2=0.91) and statistical significant slope (p < 0.0001, F=590). Decrease of blood pressure and remission of hypertensive APO did not alter significantly the EF in the same patient. From all patients, 27 had systolic heart failure (44.26%) after APO remission, the rest of 34 (55.74%) had EF-nonAPO >50%. From these patients with normal EF after APO remission, 31 (91.17%) also had normal EF-APO during hypertensive APO. None of these studied patients displayed significant acute mitral regurgitation during APO. CONCLUSIONS: Our study showed similar EF during APO and after remission of APO in patients with hypertensive APO. Normal EF in patients after remission of hypertensive APO is pointing, with good probability, to the fact that APO was due to transitory isolated diastolic heart failure because transient systolic heart failure or/and severe mitral regurgitation were rare in these patients. PMID- 19284088 TI - Genetic testing for multiple endocrine neoplasia type 2. AB - Multiple endocrine neoplasia type 2 (MEN 2) represents a complex autosomal dominant inherited syndrome characterized by occurrence of distinct proliferative disorders of endocrine tissues. The identification of RET proto-oncogene mutations in MEN 2 and FMCT has provided a precise method for identifying gene carriers. 30 subjects (9 males, 21 females, age range 11-63 years) with multiple endocrine neoplasia type 2 have been investigated from 1998 till 2006. 20 patients were considered as index cases and 10 patients were identified after a screening programme for MEN 2. Tumoral associations permitted the MEN 2A diagnosis in 21 cases, MEN 2A with cutaneous lichen amyloidosis in 6 cases and FMCT in 3 cases. We selected 22 patients from 14 families to investigate mutations in the RET proto-oncogene. In 7 subjects no mutations could be detected in the exons 10 and 11 of the RET proto-oncogene. Heterozygous missense mutations in exon 11 were found in 15 subjects consisting of three different mutations in codon 634 (TGC --> TGG, TGC --> GGC, TGC --> CGC). We conclude that our 15 patients have the most frequent mutations described in MEN 2A families. Because the testing for exons 10 and 11 is negative for other 7 patients, the remaining 13, 14, 15 and 16 exons should be sequenced in these cases. PMID- 19284089 TI - Some clinico-immunological aspects in patients with ocular myasthenia gravis associated with inflammatory bowel disease. AB - The link between inflammatory bowel disease (IBD) and ocular myasthenia gravis (OMG) is thought to be related to the production of autoantibodies. The aim of this study was to determine the incidence of some antibodies in OMG patients with IBD. Sixty-four patients with IBD and neurological symptoms were examined neurologically and immunologically (41 men and 23 women, mean age 43-5.4 years); mean duration of IBD before the diagnosis of OMG was 7.5 years. The following immunological investigations were performed: anti-acetylcholine receptor antibodies (AChR Abs) (ELISA), anti-smooth muscle antibodies (ASMA) (indirect immunofluorescence), perinuclear anti-neutrophilic cytoplasmic antibodies (p ANCA) (indirect immunofluorescence), carcinoembryonic antigen (CEA) (ELISA), Saccharomyces cerevisiae antibodies IgA and IgG (ASCA) (ELISA), peripheral lymphocyte counts, immature CD4, CD4/CD8 ratio. Five patients out of 64 with IBD presented OMG. The following antibodies were observed: anti-AChR in 4 OMG patients, ASMA in 4 OMG patients, p-ANCA in 4 OMG patients, CEA in 5 OMG patients, ASCA IgA and IgG in 3 OMG patients. Of 5 OMG patients 4 were positive for >3 antibodies. The peripheral lymphocytic counts were reduced in 5 OMG patients, a decline in CD8 cells and an increase in immature CD4 cells in 5 OMG patients, an increased CD4/CD8 ratio in 5 OMG patients were observed. The association of IBD with OMG was rare. Autoimmune dysregulation is the central defect in both MG and IBD. Further studies are required to define the nature of this association. PMID- 19284090 TI - Hyperimmunoglobulin E syndrome (Job's syndrome). AB - Hyperimmunoglobulin E (Hyper IgE) syndrome, also described as Job's syndrome, is a rare primary immunodeficiency disease characterized by recurrent skin and respiratory tract infections, chronic eczematous dermatitis, skeletal abnormalities associated with markedly elevated serum IgE levels. Variable impaired T cell function is described in this disease, but no direct association between IgE levels or clinical manifestations with these abnormalities does exist. We report a case of a patient with recurrent staphylococcal cold abscesses, eczematous dermatitis and high serum IgE levels in which the treatment with Cyclosporine A seems to have a favorable effect. PMID- 19284091 TI - Giant iliac artery aneurysm--a rare cause of hydronephrosis. AB - The presence of arterial aneurysm--an abnormal dilatation of an arterial segment due to various causes--might lead to compression effects and to various symptoms. Finding an etiology for unilateral hydronephrosis represents in some cases a real challenge for the clinician and targeted investigations must be ordered for the diagnostic approach. The abdominal ultrasound examination is one of the first imaging modalities but the abdominal computed tomography is helpful for definitive conclusions. We present a rare case of unilateral hydronephrosis due to a giant left iliac artery aneurysm in a 77-year-old male with history of peripheral artery disease. PMID- 19284092 TI - Malignancy and overdiagnosis of malignancy in Peutz Jeghers polyposis. AB - Peutz Jeghers (PJ) polyps are rare hamartomatous tumors of the gastrointestinal tract frequently associated with skin and mucosal pigmentation. Despite their benign nature there is a certain increased risk of progression to malignancy in some cases, justifying a sustained follow-up of the patients. We present 3 cases of Peutz Jeghers syndrome (PJS) diagnosed in our hospital on gastrointestinal specimens obtained by endoscopy and opened surgery. We analyzed different degrees of dysplastic changes, epithelial intussusception, association with other types of polypoid lesions and other various aspects possibly related with disease progression. Clinico-pathological correlations were made. Two of these cases were related (mother and daughter); both of them were operated in another hospital for small bowel tumors with a subsequent diagnosis of adenocarcinoma. The daughter (28 years old) was referred to our hospital for endoscopic follow-up; a small polyp of the transverse large bowel was excised by colonoscopy with a histopathologic diagnosis of PJ polyp; a careful histopathologic reevaluation of both specimens of enterectomy (slides and paraffin blocks) revealed an overdiagnosis of cancer due to the epithelial cystic dilatation and pseudoinvasion in both patients. The other case showed diagnostic changes of PJS and also various aspects of adenomatous polyps some of them with mild and moderate dysplastic changes. When a PJ polyp is diagnosed, the possibility of pseudoinvasion should be kept in mind, in order to avoid overdiagnosis of malignancy; also, due to the fact that the malignant transformation of a PJ polyp is still on debate (hamartoma-dysplasia-carcinoma sequence versus malignant transformation of an adenomatous aria of a hamartoma versus coincidental association of a digestive cancer due to genetic aberrations of PJS), all the other associated microscopic aspects of the lesion should be carefully analyzed. PMID- 19284093 TI - [Introduction of the retinal electrical biopotential into multifocal analysis]. AB - On the basis of the ISCEV standards and the author's experience, the paper describes theoretical prerequisites for multifocal electroretinography and its methodology. The variants of assessing the results, failure causes, and terminology of multifocal analysis of electroretionographic evidence are considered. PMID- 19284094 TI - [Optimization of a recording procedure of a multifocal electroretinogram]. AB - The paper presents a procedure and conditions for recording multifocal electroretinogram (mfERG), its origin, and factors influencing small-signal ERG. The optimal conditions of ERG extraction using the existing method are appraised and indications for its use in ophthalmological care defined. The results of comparing the new types of electrodes developed in Russia for ERG recording and a DTL electrode included in the ISCEV guidelines are presented. PMID- 19284095 TI - [Multifocal studies of visual functions in the diagnosis of diseases of the ocular neurosensory apparatus]. AB - The paper presents the results of using the current multifocal studies of visual functions in patients with various diseases of the retina (detachment, tapetoretinal abiotrophy, macular foramen). It provides evidence for the high correlation of changes detectable in these patients by statistical computer assisted perimetry and multifocal electroretinography. The implication of comprehensive application of these techniques to the diagnosis and differential diagnosis of ocular neurosensory apparatus diseases and to the functional monitoring and evaluation of treatment has been ascertained. PMID- 19284096 TI - [Electroretinography in regmatogenic retinal detachment in the myopic eye]. AB - The investigation studied the topographic features of retinal electrogenesis in regmatogenic retinal detachment (RRD) in the myopic eye, by using multifocal electroretinography (mfERG). Both eyes from 12 patients were used to analyze the amplitude and latency of the first negative and positive mfERG first-order kernel components averaged over the entire stimulation area, by rings, quadrants of vision fields and in the areas of the adjacent and detached retina. The myopic eye in RRD even without macular involvement showed a selective reduction in R1 amplitude when central retinal rings 1 and 2 were stimulated, which suggests that the macular region is highly sensitive to the impaired anatomic integrity of the adjoining retinal zones. Longer latency of mfERG N1 is most typical of the stimulation of the retinal macular region in recent RRD, and that is characteristic of the near and middle retinal periphery in a 1-2-month history of RRD, which may be indicative of the progression of impairments in the rod system. The reduction in P1 amplitude is most pronounced in a 1-2-month history of RRD. The findings suggest that a 1-month history is critical in the development of specific functional changes in the detached retina and mainly in its mid periphery and support a need for early (less than 4-week) surgical treatment for retinal detachment particularly in the eyes of patients with myopic changes in the eye grounds. The authors show it advisable to widely use a preliminary test protocol in poor unilateral gaze fixation (19 hexagons), which causes an increase in recording purity and, therefore, promotes the reliability of the findings. PMID- 19284097 TI - [Macular and multifocal electroretinography in the evaluation of the retinal macular region function in age-related macular degeneration]. AB - The electrogenesis of the retinal macular region versus optic coherent tomographic data was studied in different forms of age-related macular degeneration (ARMD). A macular electroretinogram (M-ERG) and mixed ERG, multifocal ERG (mf-ERG), and optical coherent tomograms were recorded in 60 patients (112 eyes) aged 57 to 82 years who had ARMD. A significant moderate correlation was established between the averaged N1 amplitude of mf-ERG in the central rings and the M-ERG a-wave to a red stimulus, as well as between the averaged latency of an mfERG P1-component and the latency of the latency of b wave and between the amplitudes of respective components. There was a significant reduction in the amplitude of the retinal macular region in confluent druzen. In focal atrophy of the retinal pigment epithelium (RPE), the amplitude decrease rate and extent varied from a significant reduction in M-ERG b-wave amplitude and a P1 decrease in the central hexagon to the significant reduction in the amplitude of M-ERG a- and b-wave components and mfERG N1 and P1 components in the foveal and parafoveal rings. M-ERG and mfERG changes in exudative ARMD were different in neuroepithelial and RPE detachments, retinal cystic changes and in varying activity of choroidal neovascularization. PMID- 19284098 TI - [Multifocal electroretinography at different stages of primary open-angle glaucoma]. AB - The paper considers the possibilities of using multifocal electroretinography (mfERG) to examine patients with primary glaucoma. Forty-seven patients underwent mfERG that established that as the disease progressed, there were reductions in the mfERG wave amplitude, when the first flash response, and in biopotential density, when the quadrant analysis was made. Biopotential changes in glaucoma can be analyzed in relation to the first flash response and a pattern response. Monitoring of changes in primary glaucoma may use mfERG. PMID- 19284099 TI - [Loal and multifocal electroretinography in the diagnosis of diabetic retinopathy]. AB - The paper gives the results of electroretinography (ERG) in 38 patients (73 eyes) with type 2 diabetes mellitus at different stages of diabetic retinopathy (including in diabetic macular edema). The reductions in the macular ERG (M-ERG) amplitude parameters and in the retinal density of bioelectrical activity and the decreased amplitude and longer latency of multifocal ERG (mfERG) N1 and P1 components evidence the involvement of the retinal macular region into the pathological process and the magnitude of these changes depends on the presence of imaged macular edema. Despite the certain limitations of mfERG, assessment of the topography of retinal bioelectrical activity in the central field of vision is an important supplement to M-ERG results and may be a prognostic criterion for the development of a pathological process and its extent. PMID- 19284100 TI - [Ratio of multifocal electroretinographic to retinal tomographic parameters in patients with suspected glaucoma]. AB - Based on the analysis of the data of a multifocal electroretinography FOK response and the retinal thickness in 38 patients with suspected glaucoma, the paper shows the inhibition of the external retinal biopotential and the lower retinal thickness in the posterior ocular pole. There is a correlation of these changes with light sensitivity alterations, as evidenced by quadrant analysis. It is stressed that monitoring of patients with suspected glaucoma must comprise a morphofunctional assessment using the current techniques. PMID- 19284101 TI - [Macular and multifocal electroretinography in the diagnosis of Stargardt's dystrophy]. AB - Retinal bioelectrical activity was studied in Stargardt's dystrophy from the data of multifocal and macular electroretinography (M-ERG) and retinal morphometry was carried out. The group of 11 patients (22 eyes) with the central form of Stargardt's dystrophy showed reductions in the retinal density of a bioelectric response of a P1 component, the amplitude of N1 and P1 components in the area of 0 to 16 degrees, as well as decreases in the amplitude of M-ERG a- and b-waves and in the retinal thickness in the macular region. The mixed Stargardt's dystrophy group was characterized by reductions in the retinal density of an electrical response of a P1 component, in the amplitude of N1 and P1 components in the entire posterior pole in the area of 0 to 32 degrees, as well as decreases in the a- and b-waves of M-ERG, photopic, scotopic, and mixed ERG; the lowered retinal thickness was found in the foveal and parafoveal regions. A statistically significant relationship was established between the morphometric indices of the volume of the central retina, the amplitude of M-ERG b-wave, and the moderate retinal density of P1 component bioelectrical activity in the central regions. PMID- 19284102 TI - [Multifocal electroretinography in ophtalmological symptoms in patients with antiphospholipid syndrome]. AB - The paper describes the first experience in using multifocal electroretinography (mfERG) in patients with antiphospholipid syndrome (APS). In the latter, thrombogenesis is mosaic in most cases so the techniques providing a spatial imaging of the pathological process is of interest in the examination of patients with this disease. These techniques include mfERG. The quadrant mfERG analysis that reveals pathological changes in relation to the vascular involvement area is of the greatest informative value in patients with APS-associated retinal vascular occlusions. PMID- 19284103 TI - [Papilledemas: clinical features in space-occupying lesions of the brain]. AB - Neuroophthalmological symptoms were analyzed in 513 patients with papilledemas in space-occupying lesions of the brain. The ophthalmoscopic features of papilledemas, the pattern of visual impairments and the developmental pattern of papilledemas were considered. Several determinants of the clinical course of papilledemas, such as their stage, the grade of malignancy, and the site of a space-occupying lesion, and a patient's age, were identified. The ability to diminish visual functions generates a need for early detection of papilledemas in patients with space-occupying lesions of the brain and for timely neurosurgical treatment in order to normalize intracranial pressure. PMID- 19284104 TI - [Potentialities of confocal microscopy in the evaluation of the cornea in the dry eye syndrome]. AB - The purpose of the investigation was to study the potentialities of confocal microscopy in the evaluation of the corneal status in the dry eye syndrome (DES). Fifty-two patients (104 eyes) were examined and divided into 2 groups: 1) 32 patients with the verified diagnosis of DES of various etiology; 2) 20 healthy volunteers (a control group). Corneal confocal microscopy was made on a Confoscan 4 device (Nidek, Japan). The investigations ascertained that there was a direct relationship between the functional parameters, the degree of severity, rather than the etiology of DES and the changes in the corneal morphological pattern. Overall, corneal changes in patients with DES primarily involved the anterior epithelium and anterior stromal layers. The non-invasive and high informative value of confocal microscopy made it possible to study at the fundamentally new level the corneal structure that is always involved in the pathological process in DES. PMID- 19284105 TI - [Photodynamic therapy with visudine in combination with intravitreal kenalog in the treatment of choroidal neovascularization]. AB - There are current tendencies in the use of photodynamic therapy (PDT) in combination with intravitreal injection of long-acting corticosteroids. Kenalog was intavitreally injected in patients with abnormal myopia (4 eyes), multifocal choroiditis (3 eyes), and age-related macular degeneration (6 eyes), and idiopathic choroidal neovascularization (CNV) (1 eye) 10-14 days before PDT. All the above cases were characterized by a severe clinical course with significantly diminished visual function, and evident exudative-hemorrhagic manifestations. A 2 5-line vision increase and bleeding resorption were observed in the patients 1-2 months after PDT. The results of optical coherent tomography suggest a significant reduction or disappearance of exudative manifestations (retinal edema, detachments of the neuroepithelium or pigment epithelium). Fluorescence angiography revealed decreased or ceased dye diffusion in the CNV areas at the late stages of the study. The findings suggest the efficiency of combined therapy for CNV that can improve visual prognosis significantly in patients after PDT. PMID- 19284106 TI - [Malignant (non-Hodgkin's) lymphoma of the organ of vision]. AB - Based of their experience and analysis of the papers available in the medical literature, the authors describe the clinical picture of malignant lymphomas of the organ of vision. They give a concept of the current classification of lymphomas and information on the specific features of their course, by taking into account the site of the process (eyelids, conjunctiva, intraocular and orbital regions) and present recommendations on their treatment. PMID- 19284107 TI - [A clinical case of pigmentary paravenous retinochoroidal atrophy]. PMID- 19284108 TI - [Phytoaeroionotherapy of psychovegetative syndrome]. AB - The objective of the study was to elucidate the possibility to use phytoaeroionotherapy as a component of combined correction of the functional state of patients with psychovegetative syndrome. The curative effect of the proposed methods was due to a decrease of the severity of manifestations of hypersympaticotony by 59-61%, a 2.5-2.7-fold reduction of pathological hyperactivity of heart meridian CV, and normalization of blood flow in the system of internal carotid and vertebral arteries. It was shown that a combination of magnetopuncture and phytoaeroionotherapeutic chronocorrection produced the best results by virtue of a 14% reduction of systolic blood pressure, decrease of the heart rate and cardiac work index by 20% and 31% respectively. The combination ofsoft-tissue manual therapy contributed to the improvement of characteristics of short-term memory by 51% and of intellectual work capacity by 50%. PMID- 19284109 TI - [The rationale for chronooptimization of rehabilitative treatment under mid latitude conditions]. AB - Rhythmic patterns in the functioning of different systems of the body should be taken into consideration when developing new therapeutic modalities intended for the improvement of efficiency of health care measures. The study of chronobiological features of the work of organ systems in patients with different diseases residing in mid-latitude regions provided a rationale for the differential approach to the prescription of medicinal preparations of natural origin in selected periods of the annual cycle. PMID- 19284110 TI - [The influence of "dry" bi-carbonate baths on the circadian profile of arterial pressure in patients who suffered acute myocardial infarction]. AB - The objective of this study was to evaluate effect of "dry" bi-carbonate baths (DBB) on the circadian profile of arterial pressure (AP) in patients who suffered acute myocardial infarction (AMI). Diurnal AP profiles were analysed in all patients after AMI. The patients comprising group 1 were treated with the use of DBB while the remaining ones received a sham treatment. 24 hour AP monitoring was performed before and after therapy. The study revealed a reduction in the frequency of hypotensive diastolic values at the daytime and in the duration of night-time hypertensive episodes under the influence of therapy with the use of DBB. Variability of systolic AP throughout 24 hours and of diastolic AP at daytime also decreased. Generally speaking, patients with elevated AP showed a more pronounced effect of DBB on the night AP profile. It is concluded that therapy with the use of DBB after AMI has beneficial effect on the clinical state of the patients and reduces the risk of cardiovascular complications. PMID- 19284111 TI - [Non-medicamentous correction of changes in the cardiovascular and respiratory systems in subjects exposed to chrysolite asbestos dust]. AB - Two physiotherapeutic complexes are proposed for the improvement of functional characteristics of the respiratory and cardiovascular systems that can be used for the non-medicamentous treatment of patients with cardiovascular diseases or concurrent broncho-pulmonary and cardiovascular pathologies. One complex is designed for the treatment of patients with asbestosis including that complicated by chronic obstructive bronchitis in the phase of remission with the well apparent signs of myocardial electric instability. The use of the other complex is indicated to patients with chronic obstructive bronchitis in the phase of exacerbation and the signs of chronic pulmonary heart associated with cardiac dysrhythmia. PMID- 19284112 TI - [Parameters of lipid peroxidation and antioxidative protection in patients with chronic pancreatitis treated by low-intensity laser therapy]. AB - The objective of this study was to evaluate the influence of low-intensity laser therapy (LILT) on the processes of lipid peroxidation (LPO) and antioxidative protection (AOP) in patients with chronic pancreatitis. A total of 78 patients aged from 36 to 77 years were treated with LILT in addition to conventional therapy; the patients of the control group (n = 40) received only medicamentous therapy. Examination of the patients in the exacerbation phase of chronic pancreatitis revealed activation of LPO processes and differently-directed shifts of AOP components. These changes are supposed to reflect variations in the activity of the inflammatory processes in the pancreas and oxidative stress on this organ. PMID- 19284113 TI - [Effect of rehabilitation on the parameters of quality of life in children with chronic gastroduodenal pathology]. AB - The objective of this study was to evaluate effect of comprehensive rehabilitation on dynamics of characteristics of the quality of life in children with chronic gastroduodenal pathology. The study included 155 children aged 14-15 years examined in the phase of exacerbation of chronic gastroduodenitis (CGD). Sixty one of these patients were given treatment in the "Detskie dyuny" children's sanatorium rehabilitation centre and the remaining 54 received traditional combined therapy services as outpatients (control group). The quality of life was ascertained based on the results of the SF-36 and WHOQOL questionnaires; the Luscher colour test, the CAH scale, and Spielberger-Khanin test anxiety inventory were used to measure the psychophysiological status of the patients. The study has demonstrated high efficacy of comprehensive rehabilitation in children during exacerbation of chronic gastroduodenal pathology that resulted in normalization of their physical and psychophysiological characteristics and improvement of the quality of life. PMID- 19284114 TI - [Diagnostic potential of chronoaximetric electrodiagnostics in the assessment of denervation-induced defects of m. soleus in patients with diabetic neuropathy]. AB - The study was designed to elucidate the informative value of chronaximetry for the quantitative assessment of denervation-induced defects of m. soleus in patients with diabetic neuropathy. Conditions are clarified under which chronaximetric measurements disagree with the position of the IT hyperbola; this discrepancy lowers substantially the informative value of these characteristics as a measure of the degree of manifestation of diabetic neuropathy. The value of chronaximetry for this purpose is markedly surpassed by that of the position of the vertex of IT hyperbola. Two approaches are proposed to the simplified and exact calculation of the position of the vertex of IT hyperbola. The length of the excitation pulse obtained by these methods (CRP6 and TGT parameters) may be used, similar to chronaximetric measurements, both for the evaluation of neuropathic changes and for the choice of optimal electrostimulation technique. PMID- 19284115 TI - [Application of low-intensity laser radiation and endotoxin-binding preparations to the treatment of female infertility]. AB - A total of 38 women of reproductive age (from 20 to 45 years) with chronic inflammatory gynecological diseases including 7 with primary and 9 with secondary infertility were examined by standard clinical, instrumental, and laboratory methods. In addition, variations of such important characteristics as serum endotoxin level and activity of antiendotoxin immunity were measured. The study has demonstrated participation of chronic aggression of endotoxins (of intestinal origin) in pathogenesis of the disorders of interest. Inclusion of the "antiendotoxic component" in the combined therapy allowed the efficacy of the treatment of chronic inflammation and female infertility to be greatly enhanced. It suggests the important (if not decisive) role of bacterial lipopolysacchardides in the pathogenetic mechanism underlying the problems considered in this study. PMID- 19284116 TI - [Transcerebral interference therapy in postoperative rehabilitation of neuro hormonal disturbances in patients who underwent partial breast resection]. AB - This study included a total of 40 patients aged between 22 and 45 (mean 31.3 +/- 1.8) years after they underwent sectoral breast resection for the removal of fibroadenoma (the diagnosis was verified by histological findings). All the patients were given transcerebral interference therapy (TCIT) one months after surgery. This therapy resulted in normalization of menstrual function in 45% of the patients, functional activity of the limbico-reticular complex in 43%, estradiol to progesterone ratio in 46%, luteinizing and folliculostimulating function of the pituitary in 54% and 35% respectively, glucocorticoid finction of the adrenal glands in 69%, and the function of the hypothalamo-pituitary-ovarian adrenal axis in 35% of the patients. The follow up ultrasound mammography 6 and 12 months after the treatment revealed clear-cut differentiation of mammary tissues. PMID- 19284117 TI - [Physiotherapeutic correction of visual functions in rehabilitation of patients after phacoemulsification of cataract with concomitant ophthalmopathology]. PMID- 19284118 TI - [Effect of superficial reflexotherapy (redox-therapy) on the psychophysiological status of athletes]. AB - The objective of this study was to evaluate the influence of superficial reflexotherapy (redox-therapy) on the psychophysiological status of athletes. It was shown to have positive effect on central and peripheral hemodynamics and beneficial action on neurovegetative regulation of the blood circulatory system. These effects depend on the choice of applicators and the duration of therapy. Superficial reflexotherapy can be used alone or in combination with physiotherapy. PMID- 19284119 TI - [Clinico-functional rationale for normobaric hypoxytherapy in rehabilitation of children and adolescents with arterial hypertension]. PMID- 19284120 TI - [Improvement of management in a sanatorium-and-spa institution]. PMID- 19284121 TI - [Evaluation of tracheal intubation conditions in morbidly obese patients: a comparison of succinylcholine and rocuronium]. AB - OBJECTIVE: To compare tracheal intubation conditions in morbidly obese patients 60 seconds after administration of succinylcholine or rocuronium doses based on real weight or ideal weight. METHOD: We evaluated patients with a body mass index (BMI) of 40 kg x m(-2) or more but no other indications of difficult-intubation risk. Induction was performed under fentanyl and propofol at doses calculated based on real weight. Patients were assigned to 1 of 4 groups for tracheal intubation. Group 1 received 1 mg of succinylcholine per kilogram of ideal weight, group 2 received 1 mg of succinylcholine per kilogram of real weight, group 3 received 0.6 mg of rocuronium per kilogram of ideal weight, and group 4 received 0.6 mg of rocuronium per kilogram of real weight. Tracheal intubation was performed 60 seconds later and intubation conditions were recorded using a clinical scoring system. RESULTS: Eighty patients with a mean BMI of 47.5 kg x m( 2) were enrolled. The difficult intubation rate was 3.75%. All patients were intubated. Laryngoscopy conditions and position and movement of vocal cords were similar in all 4 groups. Reaction to cuff inflation revealed intergroup differences; group 3 presented slight limb movements, diaphragm movement, and sustained cough for more than 10 seconds. CONCLUSION: These results suggest that administration of succinylcholine calculated based on real weight or ideal weight and rocuronium based on real weight can provide clinically acceptable conditions for tracheal intubation in morbidly obese patients with no other difficult intubation criteria. PMID- 19284122 TI - [Survey of anesthesiologists' practice in treating spontaneous aneurysmal subarachnoid hemorrhage]. AB - OBJECTIVES: To evaluate the participation of Spanish anesthesiology departments in the management of patients hospitalized for spontaneous aneurysmal subarachnoid hemorrhage. MATERIAL AND METHODS: Chiefs of anesthesiology departments of hospitals listed in the Spanish National Catalog of Hospitals of the Ministry of Health and Consumer Affairs were sent a questionnaire with 30 items covering protocols for the management of patients with spontaneous aneurysmal subarachnoid hemorrhage. Items asked about the participation of anesthesiologists during both admission and the perioperative period. RESULTS: The questionnaire was sent to 132 hospitals, of which 18 (13.6%) responded. Forty six percent of anesthesiology departments do not participate in the initial resuscitation. Only 4 reported having a protocol for treating these patients. The initial diagnosis was reportedly made by cranial computed tomography in all cases. Endovascular treatment was the most common procedure reported (66%) and it was given within the first 48 hours (66%). Basic monitoring was used more than nervous system monitoring. Total intravenous anesthesia was used for craniotomy in 53% of the hospitals and for endovascular treatment in 64%. Complications reported most often were vasospasm (100%) and hydrocephalus (69%). CONCLUSIONS: Even though few questionnaires were returned, the results reveal scarce use of protocols for the treatment of spontaneous aneurysmal subarachnoid hemorrhage by anesthesiologists. It was also evident that the participation of anesthesiology department staff in the treatment of this condition takes place almost exclusively in the intraoperative period and that the use of nervous system monitoring is scarce. Endovascular treatment is increasing in our practice settings. PMID- 19284123 TI - [Epidural analgesia versus femoral or femoral-sciatic nerve block after total knee replacement: comparison of efficacy and safety]. AB - OBJECTIVE: Pain after total knee replacement surgery is intense. The aim of this study was to compare 3 techniques for providing postoperative analgesia (epidural analgesia, femoral nerve block, and a combined femoral-sciatic nerve block) in total knee arthroplasty. MATERIAL AND METHODS: Observational study of 1550 elective primary unilateral total knee replacement operations. The safety and efficacy of the following 3 techniques were compared: epidural analgesia, femoral nerve block, and femoral-sciatic nerve block. Demographic, anesthetic, and surgical data were recorded. Study variables included pain intensity on a visual analog scale every 4 hours, need for rescue analgesia (morphine), complications and adverse events within 5 postoperative days. RESULTS: No significant differences were found in demographic, anesthetic, or surgical variables. In the first 24 hours after surgery, pain intensity was significantly less for patients who received a femoral-sciatic nerve block. The mean levels of morphine consumption in the first 96 hours after surgery were similar in the femoral sciatic nerve block group (3.18 mg) and the epidural analgesia group (3.19 mg); morphine consumption in the femoral block group was significantly higher (4.51 mg). Epidural analgesia was associated with the highest rate of complications (17%). CONCLUSIONS: A sciatic nerve block combined with a femoral nerve block attenuates pain more effectively and is associated with less postoperative morphine consumption in comparison with a femoral nerve block alone. Peripheral nerve block techniques have fewer adverse side effects than epidural analgesia. PMID- 19284124 TI - [Paresthesia and spinal anesthesia for cesarean section: comparison of patient positioning]. AB - OBJECTIVE: To determine the incidence of paresthesia during lumbar puncture performed with the patient in different positions. MATERIAL AND METHODS: A single blind prospective study of patients scheduled for elective cesarean section, randomized to 3 groups. In group 1 patients were seated in the direction of the long axis of the table, with heels resting on the table. In group 2 they were seated perpendicular to the long axis of the table, with legs hanging from the table. In group 3 they were in left lateral decubitus position. Lumbar punctures were performed with a 27-gauge Whitacre needle. RESULTS: One hundred sixty-eight patients (56 per group) were enrolled. Paresthesia occurred most often in group 3 (P = .009). We observed no differences in blood pressure after patients moved from decubitus position to the assigned position. Nor did we observe between group differences in blood pressure according to position taken during puncture. CONCLUSION: Puncture undertaken with the patient seated, heels on the table and knees slightly bent, is associated with a lower incidence of paresthesia than puncture performed with the patient seated, legs hanging from the table. Placing the patient's heels on the table requires hip flexion and leads to anterior displacement of nerve roots in the dural sac. Such displacement would increase the nerve-free zone on the posterior side of the sac, thereby decreasing the likelihood of paresthesia during lumbar puncture. A left lateral decubitus position would increase the likelihood of paresthesia, possibly because the anesthetist may inadvertently not follow the medial line when inserting the needle. PMID- 19284125 TI - [Hydroxyethyl starch to protect renal function in laparoscopic surgery]. AB - OBJECTIVE: To evaluate the effect of prehydration with hydroxyethyl starch 130/0.4 (Voluven) compared to lactated Ringer solution in laparoscopic cholecystectomy. PATIENTS AND METHODS: We performed a randomized single-blind clinical trial on patients classified as ASA 1 and 2. The exclusion criteria were hypertension, kidney failure, treatment with diuretics or other antihypertensive drugs, diabetes, and use of nonsteroidal anti-inflammatory drugs. Balanced general anesthesia with remifentanil and sevoflurane was used. The total volume of administered fluids (including prehydration) was 2 mL x kg(-1) x h(-1) fasting plus 5 mL x kg(-1) x h(-1) during surgery. Group 1 was prehydrated 30 minutes before surgery with 500 mL of lactated Ringer solution in group 1; in group 2 the same quantity of Voluven was used. Ringer solution was used in both groups to provide additional fluids. Blood pressure was kept within 20% above or below baseline values. Standard anesthetic monitoring was performed. Intraoperative diuresis and creatinine clearance were recorded. The groups were compared using the t test; a P value of 05 or less was considered significant. RESULTS: Twenty nine patients were enrolled in the study. Diuresis and creatinine clearance were significantly higher in the group that received prehydration with Voluven. The mean (SD) creatinine clearance rate was 176.44 (1433) mL x min(-1) in group 1 and 61.90 (6.6) mL x min(-1) in group 2 (P = .036). The mean volume of urine excreted was 1.71 (0.06) mL x kg(-1) x h(-1) in group 1 and 0.47 (0.02) mL x kg(-1) x h( 1) in group 2 (P = .017). CONCLUSION: Prehydration with Voluven can be an effective measure for protecting renal function against the adverse effects of pneumoperitoneum in laparoscopic surgery. PMID- 19284126 TI - [Enteral feeding of critical patients]. AB - Artificial nutrition support forms part of the basic care of critical patients. Enteral feeding has been shown to be better than total parenteral nutrition at improving morbidity (infectious complications) and reducing the length of hospital stays, number of days with mechanical ventilation, and costs. As with any other treatment, enteral feeding has associated complications and side effects which should be understood and treated in order to obtain the greatest benefit from it and reduce possible adverse effects. In this review, we attempt to provide a practical summary of the use of enteral feeding in critical patients. We cover the management of the most frequent associated complications, based on new studies and current scientific evidence. The review is intended to serve as a practice guide for the routine care of severely ill patients. PMID- 19284127 TI - [Combined regional-general anesthesia with use of the Proseal laryngeal mask during prolonged peripheral plastic surgery]. AB - The ProSeal laryngeal mask offers a better interface for controlled ventilation than does a traditional mask because of the ProSeal's improved airway seal and the possibility of draining the digestive tract. Limb reconstructive surgery is normally a long procedure involving the use of grafts or flaps located at a distance from the lesion. Regional anesthesia is therefore of limited use. We report a series of 24 adult patients who underwent peripheral plastic surgery under combined regional and general intravenous anesthesia and in whom the ProSeal mask was used. Data recorded included assessment of the airway and intubation difficulty, disease and duration of the procedure, type of regional anesthesia, respiratory and hemodynamic variables, patient satisfaction, laryngeal complications, and postoperative analgesia. All patients were adequately ventilated. One episode of bradycardia was controlled with atropine. In the immediate postoperative period, no patient presented laryngeal complications and only 1 patient reported significant pain (8 on a visual analog scale). The ProSeal mask was a good choice for use with combined regional-general anesthesia, ensuring control of the airway without causing complications. Analgesia was adequate in most cases. PMID- 19284128 TI - [Orotracheal tube ignited by an electrocautery device during tracheostomy]. AB - Endotracheal tube fire during surgery is a rare complication associated with carbon dioxide laser surgery and, less often, with electrocautery. We report a case in which tracheostomy was performed because of recurrence of a laryngeal tumor. During the procedure the endotracheal tube ignited when the lumen was opened with the electrocautery device. PMID- 19284129 TI - [Endotracheal tube placement for electromyographic monitoring during fiberoptic bronchoscopic surgery on the thyroid glands]. PMID- 19284130 TI - [Combined surgical and conservative treatment in a case of iatrogenic tracheal rupture]. PMID- 19284131 TI - [Peripheral nerve block for infrainguinal arterial bypass surgery]. PMID- 19284132 TI - [Lumbar spine arthrodesis under general anesthesia in a patient allergic to fentanyl]. PMID- 19284133 TI - [Intrathoracic gossypiboma: a case report]. PMID- 19284134 TI - [Spinal anesthesia in a patient with Angelman syndrome]. PMID- 19284135 TI - [Spatial relationships between the ultrasound transducer, the needle, and nerve structures in plexus and peripheral nerve blocks]. PMID- 19284136 TI - [Pulmonary bleeding in negative-pressure pulmonary edema]. PMID- 19284137 TI - [Intradural block in essential thrombocytosis]. PMID- 19284138 TI - [Anesthesia for the surgical removal of bilateral pheochromocytomas during the second trimester of pregnancy]. PMID- 19284139 TI - [Atypical radiologic images after upper digestive tract endoscopy]. PMID- 19284141 TI - [Not Available]. PMID- 19284140 TI - [Accidental knotting of a gastric tube inserted through a supraglottic airway device with gastric access]. PMID- 19284142 TI - [Not Available]. PMID- 19284143 TI - [Not Available]. PMID- 19284144 TI - [Not Available]. PMID- 19284145 TI - [Not Available]. PMID- 19284146 TI - [Not Available]. PMID- 19284147 TI - [Not Available]. PMID- 19284148 TI - [Not Available]. PMID- 19284150 TI - [Not Available]. PMID- 19284149 TI - [Not Available]. PMID- 19284151 TI - [Not Available]. PMID- 19284152 TI - [Not Available]. PMID- 19284153 TI - [Not Available]. PMID- 19284154 TI - [Not Available]. PMID- 19284156 TI - [Not Available]. PMID- 19284155 TI - [Not Available]. PMID- 19284157 TI - [Not Available]. PMID- 19284158 TI - Design and operation of the National Survey of Children with Special Health Care Needs, 2005-2006. AB - OBJECTIVES: This report presents the development, plan, and operation of the National Survey of Children with Special Health Care Needs (CSHCN), a module of the State and Local Area Integrated Telephone Survey, conducted by the Centers for Disease Control and Prevention's National Center for Health Statistics. This survey was designed to produce national and state-specific prevalence estimates of CSHCN, describe the types of services that they need and use, and assess aspects of the system of care for CSHCN. Funding for this survey was provided by the Matemal and Child Health Bureau, Health Resources and Services Administration. METHODS: A random-digit-dial sample of households with children younger than 18 years of age was constructed for each of the 50 states and the District of Columbia. All children in each identified household were screened for special health care needs. If CSHCN were identified in the household, a detailed interview was conducted for one randomly selected child with special health care needs. Detailed interviews were also conducted for a separate national sample of children, to generate estimates for children without special health care needs, and permit comparisons with CSHCN on all study measures. The respondents were parents or guardians who knew about the children's health and health care. RESULTS: A total of 192,083 household screening interviews were completed from April 2005 to February 2007. This resulted in 40,840 completed special-needs interviews and 6,113 completed interviews for children in the comparison (referent) sample. The weighted overall response rates were 56.1% for special needs and 50.3% for referent-sample interviews. PMID- 19284159 TI - Study of apoptosis induced by cytostatics and vegetal extracts on human endothelial cell line. AB - Angiogenesis, the biological process by which new capillaries are formed from pre existing vessels, is a tightly controlled and complex process involving several factors with both stimulating and inhibiting steps. In solid tumor growth, a specific clinical turning point is the transition to the vascular phase. Once it develops an intrinsic vascular network, a tumor grows indefinitely. Tumor angiogenesis depends mainly on the release by neoplasic cells of growth factors specific for endothelial cells (ECs), able to stimulate growth of the host blood vessels. The aim of this study was to analyze the apoptotic effect of some cytostatics, Vinblastine, Rapamycin and Doxorubicin, and vegetal extracts (called VOB) isolated and purified from Vitis sp., on human EA.hy926 endothelial cell line. In a proliferation assay using Crystal Violet, we demonstrated that Vinblastine and Rapamycin cytostatics have synergistic effect on endothelial cell line EA.hy926 growth inhibition. The inhibitory effects of Vinblastine and Doxorubicin were enhanced by VOB vegetal extracts. A combined treatment of cytostatics and VOB vegetal extracts resulted in a stronger antiproliferative effect of EA.hy926 endothelial cells. Results obtained regarding the apoptosis induced on EA.hy926 endothelial cells showed that each compound alone was able to induce a significant percent of apoptotic cells in a dose-dependent manner. PMID- 19284160 TI - Analysis of the beta-lactam resistance phenotypes of Escherichia coli. An 8-year survey conducted in Greece. AB - There have been few studies, outside of France, on the resistance of Escherichi coli to beta-lactams by means of resistance phenotypes. In the present 8-year study from a tertiary Greek hospital, a statistically significant decrease in wild-type strains was noted, with a parallel increase in strains producing penicillinase. Of the total 6,089 isolates analyzed, 62.47% had no acquired resistance mechanisms while 35.70% produced penicillinase, 0.61% cefalosporinase and 0.94% extended-spectrum beta-lactamase. No overexpression of chromosomal cephalosporinase or synthesis of inhibitor-resistant enzymes was found. A shift in the pattern of penicillinase production was noted as, in the early years of the study, intermediate- and high-level penicillinase predominated whereas, in later years, low-level penicillinase prevailed. PMID- 19284161 TI - Etiology of viral pneumopathies in patients in intensive care unit under mechanical ventilation. AB - The objective of this work was to define the etiology of viral pneumopathies at the patients from reanimation section being under mechanical ventilation, making reference to viruses with respiratory tropism, and also to Chlamydia Pneumoniae and Mycoplasma pneumoniae. The subjects were 36 patients hospitalized into Service of Medical Reanimation from CHU Caen and who needed mechanical ventilation more than 48 hours. The samples from the patients were mostly nasal aspirate, 1 bronchial aspirate and 2 tracheal aspirates. The diagnosis tests were: the test of direct immunofluorescence (DIF) from the samples (for Influenza viruses A and B, Parainfluenza 1,2,3, Adenovirus and Respiratory Syncytial Virus (RSV), inoculation on the tissue culture of diploid cells MRC5, and at the appearance of cythopatic effect specific for Herpes Simplex Virus (HSV), it was made DIF for the detection of type 1 or 2, and also there were made 6 techniques of Polymerase Chain Reaction (PCR). The results of the tests were: at admission before installing the mechanical ventilation, 6 patients presented an infection with Rhinoviruses (RV), 3 with Influenza type A, 3 with HSV type 1 and 2 with Enterovirus. After a period of time from installing the mechanical ventilation, 8 patients presented an infection with HSV typel, among who 1 presented at admission an infection with RV, and 1 patient presented at 7 days from installing the mechanical ventilation an infection with RSV, and at 16 days an infection with HSV type 1. Thus, it could be concluded that in 25% from the cases of viral pneumopathies from patients being under mechanical ventilation it was an endogen reactivation of HSV type1 and only into a single case was diagnosed initially with an infection with RSV, after that it appeared also an infection with HSV typel. PMID- 19284162 TI - Molecular detection of verotoxin-producing Escherichia coli isolates in stool samples from patients with diarrhea. AB - Despite its occurence as a commensal in the human intestine, Escherichia coli is also known as a versatile gastrointestinal pathogen. Identification of diarrheagenic E. coli (DEC) requires the accurate discrimination of pathogenic strains from commensal flora, and this is not an easy task if the diagnostic tools are inadequate. As the information regarding the relative contribution of DEC among other identifiable causes of infectious diarrhea in Romanian patients is scarce, a prospective study was conducted to evaluate the prevalence of enteropathogenic Escherichia coli (EPEC) and verotoxin-producing Escherichia coli (VTEC) isolates in the diarrheagenic stool specimens of 120 children and 270 adults. PCR-based detection of the eae, bfp, vtx1 and vtx2 genes was added to the conventional culture and slide agglutination with 12 commercial EPEC antisera and O157:H7 antisera for identifying EPEC and VTEC isolates. Even though E. coli colonies belonging to traditional EPEC serogroups were isolated from 35 children and 17 adults, only the isolates recovered from 16 children and 2 adults harboured at least one of the targeted pathogenicity-associated genes. The children shedding EPEC outnumbered the adults (16.7% vs. 7.4%). Based on the virulence genotype identified, the prevalence of atypical EPEC (eae+) was higher than of typical EPEC (eae+ bfpA+), and typical EPEC identification was restricted to children. Owing to the molecular analysis 5 children and 10 adults that could have been overlooked by the routine microbiological investigation were diagnosed as infected with VTEC. Considering that none of the screened stool specimens was positive for E. coli O157:H7, this study reports for the first time the presence of VTEC nonO157 in local patients with diarrhea. Our results bring evidence that both EPEC and VTEC isolates are circulating as agents of local sporadic cases of human diarrhea. Further studies are needed to evaluate the contribution of DEC to the human disease burden in Romania, based on improved diagnostic tools targeting the main virulence traits of E. coli clinical isolates. PMID- 19284163 TI - Characteristics of Romanian fluoroquinolone-resistant human clinical Escherichia coli isolates. AB - Alarming progressive increase in the prevalence of antimicrobial resistance in Escherichia coli has been documented worldwide. Previous studies have suggested that many E. coli clinical isolates are actually low-virulence opportunists whose success derives more from antibiotic resistance than from pathogenic capability. The co-existence of ESBL production and fluoroquinolone resistance was reported as a major therapeutic challenge for E. coli infections. Considering the sparse information regarding the genetic background of virulence and antibiotic resistance of local isolates, a collection of ciprofloxacin-resistant E. coli isolates from human extraintestinal specimens was analyzed using PCR, PCR sequencing, and PFGE, in order to clarify some aspects regarding their mechanisms of antimicrobial resistance, phylogenetic origin, the content of virulence encoding determinants, and clonal relatedness. The tested fluoroquinolone resistant E. coli (FQREC) isolates, which displayed genetic heterogeneity, carried double mutations in the QRDR of gyrA previously described, which could explain their high resistance to ciprofloxacin. More than half of them (69%) possessed group 1 blaCTX. like genes, and with one exception, all these isolates were ESBL producers. The FQREC isolates belonging to non B2 phylogenetic groups outnumbered the isolates derived from B2 group (60 versus 27 isolates), and their overall content of virulence-encoding genes (fim, pap, sfa/foc, afa, hly, cnf and aer) was reduced. Regardless of the phylogenetic origin, the most prevalent virulence-associated genes possessed by the FQREC isolates were aer and fim determinants, while none of these isolates carried hly and cnf genes. In the case of weakened patients, the E. coli isolates do not need a robust virulence repertoire in order to overcome the host defense systems. The co-resistance of many FQREC isolates to extended-spectrum cephalosporins may provide a substantial advantage to their survival and spreading within the hospital environment. PMID- 19284164 TI - Antibacterial antibodies for some enterobacteria in sera of patients with reactive arthritis and other rheumatoid diseases. AB - The presence of the specific antibodies for some enterobacteria--Yersinia, Salmonella and Shigella was investigated in patients hospitalized in the period 2000-2007 with reactive arthritis and other rheumatoid diseases. The antibacterial antibodies in the diagnosis titres were found in 172 out of 1102 (15.6%) patients. Increased antibodies levels to Yersinia were detected in the sera from 113 (65.7%) of the 172 patients, for Shigella in 47 (27.3%) and for Salmonella in 12 (7.0%) cases. From all serologically positive patients 126 (73.2 %) had clinical diagnosis of reactive arthritis and 46 (26.7%) other rheumatoid diseases (ankylosing spondilytis, Reiter's syndrome, sacroilitis). The most serologically positive cases (63.9%) were of middle-age (30-50 years). There were no significant differences between sexes among serologically positive cases. PMID- 19284165 TI - Optimization of bacteriocin production by Lactobacillus plantarum AA135. AB - Growth of Lactobacillus plantarum AA135 in MRS broth at 30 degrees C yielded maximal plantaricin AA13 activity of 10000 AU/ml at pH 3.8 after 16 hours. Maximal activity (10000 AU/ml) of plantaricin AA13 was recorded in MRS broth at initial pH of 5 or 5.5. Optimal production (10000 AU/ml) was recorded in the presence of tryptone (20 g/l), a combination of tryptone plus meat extract (1:0.6) or tryptone with yeast extract (1:0.6) as sole nitrogen source. Growth of Lactobacillus plantarum AA135 in the presence of 20 g/l glucose yielded bacteriocin activity of 10000 AU/ml. Moreover, when 40 g/l glucose was added, the plantaricin AA13 levels doubled to 20000 AU/ml. Concentrations of 5, 10 and 20 g/l K2HPO4 lowered bacteriocin activity by 60%. Supplementing MRS with 1 g/l or more glycerol repressed bacteriocin production. Maximal activity of 20000 AU/ml was recorded in MRS supplemented with Vitamin DL-6,8-thioctic acid. PMID- 19284166 TI - Novel antipathogenic strategies against adherent enterobacterial strains isolated from the hospital environment. AB - The emergence of the bacterial antibiotic multi-resistance made more and more stringent the developing of new anti-microbial strategies. The purpose of the present study was to investigate the antimicrobial potential of six (6) newly synthesized chemical compounds (derivating from phenantroline and dimethylguanin copper complex combinations) versus 97 enterobacterial strains isolated from the hospital environment. The qualitative screening of the antimicrobial activity of the chemical compounds was performed by an adapted diffusion method. The minimal inhibitory concentrations (MIC) of the active chemical compounds were established by Mueller Hinton broth microdillution method. The tested chemical compounds were also tested for their ability to inhibit microbial adherence and biofilm development on inert substrata by a simple microtiter method. All six chemical compounds proved to have antimicrobial activity versus the most of the tested strains, the phenantroline derivatives exhibiting higher antimicrobial activity than the dimethylguanidine-copper complex combinations. The subinhibitory concentrations of the tested chemical products slightly inhibited the adherence ability of the bacterial strains to the inert substratum. Our results demonstrated that phenantroline derivatives may represent a new strategy of antimicrobial treatment, simultaneously with the bactericidal effect, the subinhibitory concentrations of these newly synthesized chemical compounds decreasing the adherence ability of bacteria to the inert substratum. PMID- 19284167 TI - A "hospital patient" in Ireland. PMID- 19284168 TI - Quality improvement in primary care: ethical issues explored. AB - PURPOSE: Quality improvement (QI) processes in family medicine are becoming increasingly complex. Their influence on the organisation of the sector and on the daily work processes is profound and increasing. The literature indicates that many ethical issues are arising from QI work. Therefore this paper aims to identify the experiences of professionals involved in planning and performing QI programmes in European family medicine on the ethical implications involved in those processes. DESIGN/METHODOLOGY/APPROACH: Four focus groups were carried out with 29 general practitioners (GPs) and administrators of general practice quality work in Europe. Two focus groups comprised EQuiP members and two focus groups comprised attendees to an invitational conference on QI in family medicine held by EQuiP in Barcelona in November 2006. FINDINGS: Four overarching themes were identified, including implications of using patient data, prioritising QI projects, issues surrounding the ethical approval dilemma and the impact of QI. Each theme was accompanied by an identified solution. PRACTICAL IMPLICATIONS: Prioritising is necessary and in doing that GPs should ensure that a variety of work is conducted so that some patient groups are not neglected. Transparency and flexibility on various levels is necessary to avoid harmful consequences of QI in terms of bureaucratisation, increased workload and burnout on part of the GP and harmful effects on the doctor-patient relationship. There is a need to address the system of approval for national QI programmes and QI projects utilising more sophisticated methodologies. ORIGINALITY/VALUE: This study provides data from GPs who are experienced quality improvers across 17 countries. Many ethical issues were identified and it was possible to clearly map the themes and their relationships and to summarise the identified solutions from an international perspective. PMID- 19284169 TI - Developing, testing and applying instruments for measuring rising dependency acuity's impact on ward staffing and quality. AB - PURPOSE: This paper aims to explains how relatively simple nurse staffing formulas from "best practice" ward dependency-acuity data can be used for nursing workforce planning and development. DESIGN/METHODOLOGY/APPROACH: The paper combines literature, detailed ward surveys, workshop and expert group/stakeholder information to generate and test care levels/nurse multipliers for setting ward establishments. FINDINGS: The paper finds that professional-judgement based ward staffing can be abandoned, while complex acuity-quality, timed-task and regression-based nurse staffing algorithms for setting ward establishments may be unnecessary since the new multipliers, underpinned by robust validity and reliability testing, seem to be remarkably accurate nurse-staffing determiners at a fraction of the cost. RESEARCH LIMITATIONS/IMPLICATIONS: As care levels and multipliers stand they are suitable only for UK National Health Service acute wards. Primary care, mental health, learning disability and other specialist group care levels and multipliers need developing. PRACTICAL IMPLICATIONS: Users, at a minimum, can adopt care level data and multiplier staffing recommendations for benchmarking purposes. Ultimately, the algorithms can be used to: adjust ward establishments according to workload; or set staffing for new, inpatient services. ORIGINALITY/VALUE: The paper offers a simple system for assessing patients' nursing needs and setting ward staffing accordingly. PMID- 19284170 TI - Factors affecting performance of hospital nurses in Riyadh Region, Saudi Arabia. AB - PURPOSE: The purpose of this paper is to identify factors influencing performance of hospital nurses in Riyadh Region, Saudi Arabia. Specific objectives were to estimate self-reported performance, and determine whether differences in employee demographics, job satisfaction, and organizational commitment, influenced performance. DESIGN/METHODOLOGY/APPROACH: In total, 15 hospitals were randomly selected. The questionnaire was sent to all nurses (1,834) in these facilities and 923 nurses responded. Statistical analysis included correlation, t-test, and regression analysis. FINDINGS: The study finds that job performance is positively correlated with organizational commitment, job satisfaction and personal and professional variables. Both job satisfaction and organizational commitment are strong predictors of nurses' performance. Job performance is positively related to some personal factors, including years of experience, nationality, gender, and marital status. Level of education is negatively related to performance. RESEARCH LIMITATIONS/IMPLICATIONS: The findings of this study have a limited generalisability due to the fact that all measures used are based on self reports. Future research may be directed to other objective measures of performance. PRACTICAL IMPLICATIONS: Emphasis should be placed on effective supervision, empowerment, and a better reward system. Cultural diversity is a reality for most health organizations in Saudi Arabia; therefore, they need to adopt effective human resources strategies that aim to improve commitment and retention of qualified workers, and build a high performance organizational culture based on empowerment, open communication, and appreciation of impact of national culture on work attitudes. ORIGINALITY/VALUE: This study fulfills a research gap in the area of nursing performance, and its relationship with work attitudes in Saudi Arabia. The paper also highlights the impact of national culture on job performance and work attitude among nurses in Saudi Arabia, and other countries facing the issue of multi-national work force. PMID- 19284171 TI - Protocols, policy directives and choice provision: UK midwives' views. AB - PURPOSE: Within maternity hospitals midwives are expected to follow the protocol driven culture and orders issued by senior staff. Simultaneously, midwives are expected to follow social policy documents and the Midwives Rules and Standards that advocate choice provision for childbearing women. Quality assurors and auditors of clinical practice need to be aware that these two directives sometimes clash. Allegiance to a hierarchical system driven by protocols and orders from the top down, at the same time as providing "woman-centred" care is often unattainable. In order for a midwife to action the woman's choice, resourceful thinking may be required. This paper aims to examine this issue. DESIGN/ METHODOLOGY/APPROACH: A descriptive interview study set out to discover strategies which midwives use to resolve conflict produced from competing directives. An appraisal of 20 midwives' views were gained from semi-structured interviews conducted in seven maternity units in the UK. Taking a post-positivist approach, inductive thematic analysis was used to interpret the data. FINDINGS: Three main categories represented resourceful ways of pleasing both authority and the childbearing woman. Midwives occasionally: are economical with the truth; circumvent face-to-face confrontation with senior staff; and persuade women to refuse what they perceive are unnecessary and invasive interventions. ORIGINALITY/VALUE: This paper offers unique insights into methods that midwives use to resolve conflicts in direction issued by management. It is important that auditors are aware that midwives sometimes struggle to support the preferences of healthy childbearing women. This reduces job satisfaction, delivery of care and consequently requires address. PMID- 19284172 TI - RFID in the healthcare supply chain: usage and application. AB - PURPOSE: The purposes of this study are to first, determine the most efficient and cost effective portions of the healthcare supply chain in which radio frequency identification devices (RFID) can be implemented. Second, provide specific examples of RFID implementation and show how these business applications will add to the effectiveness of the healthcare supply chain. And third, to describe the current state of RFID technology and to give practical information for managers in the healthcare sector to make sound decisions about the possible implementation of RFID technology within their organizations. DESIGN/METHODOLOGY/APPROACH: Healthcare industry literature was reviewed and examples of specific instances of RFID implementation were examined using an integrated simulation model developed with Excel, @Risk and Visio software tools. FINDINGS: Analysis showed that the cost of implementing current RFID technology is too expensive for broad and sweeping implementation within the healthcare sector at this time. However, several example applications have been identified in which this technology can be effectively leveraged in a cost-effective way. PRACTICAL IMPLICATIONS: This study shows that RFID technology has come a long way in the recent past and has potential to improve healthcare sector productivity and efficiency. Implementation by large companies such as Wal-mart has helped to make the technology become much more economical in its per unit cost as well as its supporting equipment and training costs. ORIGINALITY/VALUE: The originality of this study lies in the idea that few practical and pragmatic approaches have been taken within the academic field of study for the implementation of RFID into the healthcare supply chain. Much of the research has focused on specific companies or portions of the supply chain and not the entire supply chain. Also, many of the papers have discussed the future of the supply chain that is heavily dependent on advances in RFID technology. A few viable applications of how RFID technology can be implemented in the healthcare supply chain are presented and how the current state of technology limits the broad use and implementation of this technology in the healthcare industry. PMID- 19284173 TI - Data mining in pharma sector: benefits. AB - PURPOSE: The amount of data getting generated in any sector at present is enormous. The information flow in the pharma industry is huge. Pharma firms are progressing into increased technology-enabled products and services. Data mining, which is knowledge discovery from large sets of data, helps pharma firms to discover patterns in improving the quality of drug discovery and delivery methods. The paper aims to present how data mining is useful in the pharma industry, how its techniques can yield good results in pharma sector, and to show how data mining can really enhance in making decisions using pharmaceutical data. DESIGN/METHODOLOGY/APPROACH: This conceptual paper is written based on secondary study, research and observations from magazines, reports and notes. The author has listed the types of patterns that can be discovered using data mining in pharma data. FINDINGS: The paper shows how data mining is useful in the pharma industry and how its techniques can yield good results in pharma sector. RESEARCH LIMITATIONS/IMPLICATIONS: Although much work can be produced for discovering knowledge in pharma data using data mining, the paper is limited to conceptualizing the ideas and view points at this stage; future work may include applying data mining techniques to pharma data based on primary research using the available, famous significant data mining tools. ORIGINALITY/VALUE: Research papers and conceptual papers related to data mining in Pharma industry are rare; this is the motivation for the paper. PMID- 19284174 TI - An assessment of diagnostic efficiency by Taguchi/DEA methods. AB - PURPOSE: The aim of this paper is to propose a new, objective and consistent method for the calculation of the diagnostic efficiency in medical applications. DESIGN/METHODOLOGY/APPROACH: In this study, a hybrid method of Taguchi and DEA is proposed. This method reflects the diversity of inputs and outputs by incorporating the stepwise application of sensitivity, specificity, leveling threshold, and efficiency score. A hypothetical case study is given which involves eight readers of X-ray films in clinical radiology. FINDINGS: The selected pairs of sensitivity and specificity yielded two efficient readers. After super efficiency analysis, Reader 6 is found to be the most efficient reader. ORIGINALITY/VALUE: The paper presents a new, objective and consistent method for the calculation of the diagnostic efficiency in medical applications. PMID- 19284175 TI - Autism: is it all in the head? PMID- 19284176 TI - The doctor's dilemma: healthcare reform and integrative medicine. PMID- 19284177 TI - Systematic reviews and meta-analyses support the efficacy of numerous popular herbs and phytomedicines. PMID- 19284178 TI - The right order of things: peeling the onion of chronic disease. PMID- 19284179 TI - Does valerian improve sleepiness and symptom severity in people with restless legs syndrome? AB - OBJECTIVE: To compare the effects of 800 mg of valerian with a placebo on sleep quality and symptom severity in people with restless legs syndrome (RLS). METHODS: A prospective, triple-blinded, randomized, placebo-controlled, parallel design was used to compare the efficacy of valerian with placebo on sleep quality and symptom severity in patients with RLS. Thirty-seven participants were randomly assigned to receive 800 mg of valerian or placebo for 8 weeks. The primary outcome of sleep was sleep quality with secondary outcomes including sleepiness and RLS symptom severity. RESULTS: Data were collected at baseline and 8 weeks comparing use of valerian and placebo on sleep disturbances (Pittsburgh Sleep Quality Index and Epworth Sleepiness Scale) and severity of RLS symptoms (International RLS Symptom Severity Scale) from 37 participants aged 36 to 65 years. Both groups reported improvement in RLS symptom severity and sleep. In a nested analysis comparing sleepy vs nonsleepy participants who received 800 mg ofvalerian (n=17), significant differences before and after treatment were found in sleepiness (P=.01) and RLS symptoms (P=.02). A strong positive association between changes in sleepiness and RLS symptom severity was found (P=.006). CONCLUSIONS: The results of this study suggest that the use of 800 mg of valerian for 8 weeks improves symptoms of RLS and decreases daytime sleepiness in patients that report an Epworth Sleepiness Scale (ESS) score of 10 or greater. Valerian may be an alternative treatment for the symptom management ofRLS with positive health outcomes and improved quality of life. PMID- 19284180 TI - Cranberry constituents affect fructosyltransferase expression in Streptococcus mutans. AB - CONTEXT: Cranberry juice has long been recognized in folk medicine as a therapeutic agent, mainly in urinary tract infections. Its proposed mechanism of action is antiadhesion of bacteria. OBJECTIVE: Investigation of the potential antiadhesion effect of nondialyzed material of cranberry (NDM) via its influence on secretion, gene expression, and promoter activity of the fructosyltransferase (FTF), which is among the extracellular enzymes associated with dental biofilm formation and pathogenesis of oral bacteria. MAIN OUTCOME MEASURES: Secretion of FTF from Streptococcus mutans, in the presence of NDM, was measured by immunoblotting and confocal scanning laser microscopy. Its influence on ftf gene expression was determined by reverse transcription followed by real-time RT-PCR. The luciferase assay was used to detect bioluminescence expressed by the ftf promoter activity of bacteria exposed to NDM. RESULTS: NDM at concentrations between 0.2/mL and 1mg/mL significantly (P<.05) decreased secretion of extracellular FTF, as well as down-regulated ftf expression in a dose-dependent manner. NDM also markedly reduced the luciferase activity under the ftf promoter. PMID- 19284181 TI - Relative bioavailability comparison of different coenzyme Q10 formulations with a novel delivery system. AB - Commercial coenzyme Q10 (CoQ10, ubiquinone) formulations are often of poor intestinal absorption. The relative bioavailability of CoQ10 has been shown in National Institutes of Health-funded clinical trials to be increased by its delivery system. We investigated the bioavailability of a new CoQ10 formulation based on a new and patented technology, VESIsorb, with 3 other commercially available CoQ10 products, an oil-based formulation and 2 solubilizates. This new CoQ10 formulation (commercially branded CoQsource) is a lipid-based formulation that naturally self-assembles on contact with an aqueous phase into an association colloid delivery system (hereafter "colloidal-Q10"). Twenty healthy male and female subjects participated in a double-blind, comparative (parallel design), controlled, single-dose (120 mg) bioavailability study. Plasma concentration of CoQ10 was determined at baseline and at various intervals after administration over a 24-hour period. To compare bioavailability, maximum concentration (Cmax) and area under curve from 0 to 10 hours (AUC(0-10h)) were assessed. The kinetic profiles of all CoQ10 preparations revealed a 1-peak plasma concentration-time course. Highest Cmax values were seen after colloidal-Q10 administration. Colloidal-Q10 not only had the highest plasma concentration levels after 1 hour, but it continued to increase before reaching Cmax at about 4 hours. The plasma concentration of colloidal-Q10 remained well above the levels of the 3 other products throughout the 24-hour period. The relative bioavailability calculated using the AUC(0-10h) values was also the highest for colloidal-Q10; the AUC(0-10h) values were 30.6, 6.1, 4.9, and 10.7 microg/mL*h for colloidal-Q10, solubilizate 1, the oil-based formulation, and solubilizate 2, respectively. Differences in Cmax and AUC between colloidal-Q10 and the 3 other formulations were statistically significant. In summary, the data presented suggests that colloidal-Q10 improves the enteral absorption and the bioavailability of CoQ10 in humans. PMID- 19284182 TI - Metabolic cardiology: the missing link in cardiovascular disease. AB - The importance of supporting energy production in heart cells and the preservation of the mitochondria in these cells will be the focus of a new frontier in cardiovascular prevention, treatment, and management. Many physicians are not trained to look at heart disease in terms of cellular biochemistry; therefore, the challenge in any metabolic cardiology discussion is in taking the conversation from the "bench to the bedside." An understanding of the vital role that adenosine triphosphate (ATP) plays in the heart is critical for any physician or clinician considering therapeutic options that support ATP production and turnover in jeopardized cardiac muscle cells. Metabolic therapies that help cardiomyocytes meet their absolute need for ATP fulfill a major clinical challenge of preserving pulsatile cardiac function while maintaining cell and tissue viability. D-ribose, L-carnitine, and coenzyme Q10 work in synergy to help the ischemic or hypoxic heart preserve its energy charge. This article introduces how ATP, diastolic heart function, and metabolic support help maintain cardiac energy by preserving ATP substrates. Part 2 will investigate an in-depth biochemical discussion of congestive heart failure with physiologic, pathophysiologic, and treatment considerations. PMID- 19284183 TI - Effects of dried cruciferous powder on raising 2/16 hydroxyestrogen ratios in premenopausal women. PMID- 19284184 TI - A possible central mechanism in autism spectrum disorders, part 3: the role of excitotoxin food additives and the synergistic effects of other environmental toxins. AB - There is compelling evidence from a multitude of studies of various design indicating that foodborne excitotoxin additives can elevate blood and brain glutamate to levels known to cause neurodegeneration and in the developing brain, abnormal connectivity. Excitotoxins are also secreted by microglial activation when they are in an activated state. Recent studies, discussed in part 1 of this article, indicate that chronic microglial activation is common in the autistic brain. The interaction between excitotoxins, free radicals, lipid peroxidation products, inflammatory cytokines, and disruption of neuronal calcium homeostasis can result in brain changes suggestive of the pathological findings in cases of autism spectrum disorders. In addition, a number of environmental neurotoxins, such as fluoride, lead, cadmium, and aluminum, can result in these pathological and biochemical changes. PMID- 19284186 TI - What kind of dentist are you? PMID- 19284185 TI - Jason Hao, DOM: pioneering the use of scalp acupuncture to transform healing. Interview by Frank Lampe and Suzanne Snyder. PMID- 19284187 TI - Fosamax and osteonecrosis of the jaw--important questions and answers. PMID- 19284188 TI - Patient communication for restorative dentistry using a customized written document. AB - Effective communication with patients allows them to understand their problems and what causes them. At times, patients also may need some motivation to initiate therapy. From a marketing standpoint, anything that can be done in a professional manner to enhance the patient's perceived image of the dentist and practice will be beneficial to both the patient and the dentist. The type of letter described in this column is one example of how to accomplish all of the goals. PMID- 19284189 TI - Polishing porcelain--is it even possible intraorally? PMID- 19284190 TI - Prosthetic implant challenges. PMID- 19284191 TI - Nonsurgical management of interdental papillae loss following extraction of anterior teeth. AB - When a tooth is extracted, the interproximal papilla recedes and the buccal bone collapses; as a result, restoring an edentulous space in the esthetic zone becomes a challenge. Various surgical reconstructions can be performed with different levels of risk and predictability. This article examines three nonsurgical approaches utilized to manage the same problem. One option involves placing a temporary bridge with an ovate pontic at the time of extraction to support the proximal papillae, the facial soft tissue, and the healing gingival tissue. If some time has passed since the extraction, the soft tissue can be sculpted with radiosurgery (or a laser) before an ovate pontic is used. In some instances, tissue-colored porcelain may help to achieve optimal esthetics. PMID- 19284192 TI - Benefits of preheating resin composite before placement. AB - Resin composite can be relatively stiff, difficult to express from the compule/syringe, and challenging to place. Research has shown that heating the material prior to use not only decreases its viscosity but also improves its physical properties. This article discusses the effect of preheating composite and discusses a device designed to control the preheating process. PMID- 19284193 TI - Long-term progression of a solitary bone cyst: a case report. AB - This article presents a case involving a solitary bone cyst in a 16-year-old boy who had a history of trauma in the affected region. Radiographs confirmed the presence of the cyst three years before treatment. A series of similarly produced panoramic radiographs demonstrated a chronologic picture of a solitary bone cyst, representing a lesion that was progressive rather than static. The patient was treated with curettage and experienced an uneventful recovery. The patient did not show any signs or symptoms of recurrence three years after treatment. PMID- 19284194 TI - Vasovagal syncope in children and adolescents. AB - Vasovagal syncope is not an uncommon event in children and adolescents; however, this fact is not widely known among the dental community. Although there is little information about the incidence of vasovagal syncope occurring among children and adolescents in the dental setting, dentists should always take care to eliminate factors that can predispose a patient to a vasovagal reaction. The dental team should be familiar with the emergency protocols for a vasovagal syncope episode. For this study, the author conducted a MEDLINE search using the key terms "vasovagal syncope", "syncope in children", "syncope treatments", and "syncope and dentistry" to define the pathophysiology, incidence, treatment options, and dental implications of vasovagal syncope. The articles included those published in English and in peer-reviewed journals, in addition to the most recent medical and dental textbooks as they related to syncope. PMID- 19284195 TI - Death by magnets: an unrecognized danger in dentistry. AB - Small, powerful magnets have been used in dentistry for a variety of orthodontic and prosthodontic treatments; however, if these magnets become loose and are swallowed, patients are at risk for bowel obstruction and perforation. This article makes recommendations concerning close inspection and reinforcement of intraoral devices that contain magnets. PMID- 19284196 TI - Influence of calcium hydroxide intracanal medication on the sealing ability of warm gutta-percha. AB - This in vitro study sought to evaluate the influence of calcium hydroxide on the sealing ability of vertically condensed warm gutta-percha filling. Sixty single rooted teeth were instrumented with a rotary system and divided randomly into two groups of 26 teeth each; the remaining eight teeth were divided into two groups of four to serve as negative and positive controls. The canals in the first group were filled with calcium hydroxide paste while the canals in the second group received no medication. Seven days after incubation, root canal dressings were removed by irrigation, using 5.25% sodium hypochlorite associated with ultrasonics and 50% citric acid. All specimens were filled using the warm gutta percha vertical compaction technique, placed in test tubes containing India ink, and centrifuged at 30 G for five minutes; at that point, the dye penetration level was scored. Under the conditions of this study, the calcium hydroxide group showed significantly greater dye penetration than the non-medicated group. PMID- 19284197 TI - Atypical odontalgia: a case report. AB - Diagnosis and treatment of orofacial pain is not uncommon; however, reaching a definitive diagnosis in these cases can be a complex challenge. Dentists are most likely to face this situation, because persistent and chronic pain is more common in the head and neck region than in any other part of the body. However, the complexities and diagnostic challenges mean that misdiagnosing neuropathic pain is common. This article presents a case of atypical odontalgia and illustrates the complexities involved when diagnosing the condition. PMID- 19284198 TI - A Cochrane systematic review finds no reliable evidence for different management options for the fractured edentulous atrophic mandible. AB - Cochrane reviews provide high-quality, reliable, and up-to-date evidence concerning the effects of health care interventions. This review sought to provide reliable evidence regarding the effectiveness of any interventions (either open or closed) that could be used to manage a fracture of the edentulous mandible. A sensitive search strategy was developed and used across several databases (including the Cochrane Oral Health Group Trials Register, the Cochrane Central Register of Controlled Trials, MEDLINE, and EMBASE) to identify randomized controlled trials and compare treatments for the management of fractured edentulous atrophic mandibles. Eligible studies were screened independently by two review authors and methodological quality was assessed based on criteria defined by the Cochrane Collaboration. Based on this review, there is inadequate evidence at present to confirm the effectiveness of a single approach for managing fractured atrophic edentulous mandibles; clinicians should continue basing treatment decisions on experience until high-level evidence is available. PMID- 19284199 TI - The patient taking antiplatelet drugs: a review with dental management considerations. AB - Antiplatelet drugs are used in clinical practice to prevent the adverse sequelae of thromboses in atherosclerotic arteries of the heart, brain, and limbs and in the veins and heart chambers. They have diverse mechanisms of action, half-lives, and pharmacodynamic effects. A major concern among dental health care providers is the potential for excessive bleeding after invasive dental procedures. This article reviews the current antiplatelet agents used for managing cardiovascular and cerebrovascular diseases and suggests how patients taking these agents may be managed when invasive dental procedures are planned. PMID- 19284200 TI - Effect of long-term water storage on etch-and-rinse and self-etching resin-dentin bond strengths. AB - This study sought to evaluate the long-term bond strength of etch-and-rinse and self-etch adhesive systems to dentin after one year of water storage. Crown fragments from the buccal surface of extracted bovine incisors were ground flat to expose dentin surfaces. Four etch-and-rinse and two self-etch bonding agents were used according to manufacturers' instructions. Bonded specimens were stored in water for either 24 hours or one year at 37 delete C. After elapsed storage times, specimens were tested for shear bond strength (SBS) at 0.5 mm/min. Data were analyzed by ANOVA and Tukey's tests (p < 0.05). Both evaluated factors (adhesive system and storage time) and their interactions were statistically significant. Single Bond produced the highest SBS regardless of the water degradation period. One-year water storage reduced the SBS of Prime & Bond NT and One-Up Bond F significantly; the other adhesives performed similarly. Long-term bond strengths of etch-and-rinse and self-etch adhesive systems are susceptible to hygroscopic and hydrolytic effects to varying extents, depending on their chemistry and structure. The presence of monomers with different properties might induce preferential degradation of specific adhesive polymer systems. PMID- 19284201 TI - A clinical pathway for complete immediate denture therapy: successful prosthetic management for hopeless dentitions. AB - This article presents a rationale for utilizing complete immediate denture therapy, highlighting various advantages, disadvantages, and contraindications for this treatment. Jerbi's technique for presurgical cast carving is reviewed and a unique case report is detailed, involving a 22-year-old patient whose hopelessly debilitated dentition was treated by full-mouth extraction and placement of a maxillary and mandibular complete immediate denture. In addition, a brief review of changing dietary patterns and social habits is presented in relation to the recent escalation of rampant dental caries in youth and young adult patient populations. PMID- 19284202 TI - Incidental radiographic finding. Simple bone cyst. PMID- 19284203 TI - Multiple unerupted teeth. Cleidocranial dysplasia. PMID- 19284227 TI - Predictors of hearing preservation after stereotactic radiosurgery for acoustic neuroma. AB - OBJECT: Many patients with acoustic neuromas (ANs) have hearing function at diagnosis and desire to maintain it. To date, radiosurgical techniques have been focused on conformal irradiation of the tumor mass, with less attention to inner ear structures for which there was scant radiobiological information. The authors of this study evaluated tumor control and hearing preservation as they relate to tumor volume, imaging characteristics, and nerve and cochlear radiation dose following stereotactic radiosurgery (SRS) using the Gamma Knife. METHODS: Seventy seven patients with ANs had serviceable hearing (Gardner-Robertson [GR] Class I or II) and underwent SRS between 2004 and 2007. This interval reflected more recent measurements of inner ear dosimetry during the authors' 21-year experience. The median patient age was 52 years (range 22-82 years). No patient had undergone any prior treatment for the ANs. The median tumor volume was 0.75 cm(3) (range 0.07-7.7 cm(3)), and the median radiation dose to the tumor margin was 12.5 Gy (range 12-13 Gy). At diagnosis, a greater distance from the lateral tumor to the end of the internal auditory canal correlated with better hearing function. RESULTS: At a median of 20 months after SRS, no patient required any other additional treatment. Serviceable hearing was preserved in 71% of all patients and in 89% (46 patients) of those with GR Class I hearing. Significant prognostic factors for maintaining the same GR class included (all pre-SRS) GR Class I hearing, a speech discrimination score (SDS) >or= 80%, a pure tone average (PTA) < 20 dB, and a patient age < 60 years. Significant prognostic factors for serviceable hearing preservation were (all pre-SRS) GR Class I hearing, an SDS >or= 80%, a PTA < 20 dB, a patient age < 60 years, an intracanalicular tumor location, and a tumor volume < 0.75 cm(3). Patients who received a radiation dose of < 4.2 Gy to the central cochlea had significantly better hearing preservation of the same GR class. Twelve of 12 patients < 60 years of age who had received a cochlear dose < 4.2 Gy retained serviceable hearing at 2 years post-SRS. CONCLUSIONS: As currently practiced, SRS with the Gamma Knife preserves serviceable hearing in the majority of patients. Tumor volume and anatomy relate to the hearing level before radiosurgery and influence technique. A low radiosurgical dose to the cochlea enhances hearing preservation. PMID- 19284228 TI - Predictors of patency of excimer laser-assisted nonocclusive extracranial-to intracranial bypasses. AB - OBJECT: Excimer laser-assisted nonocclusive anastomosis (ELANA) is a technique that can be used for extracranial-to-intracranial (EC-IC) bypasses, without the necessity of temporary occlusion of the donor or recipient artery. Information on predictors of patency of EC-IC bypasses in general and the ELANA bypass in particular is sparse. The authors studied 159 ELANA EC-IC bypasses to find predictors of patency. METHODS: From a prospective database of patients who underwent EC-IC bypass surgery, 143 consecutive patients who underwent a total of 159 ELANA bypasses were studied. The associations of patient characteristics, surgical aspects, and technical aspects specific to the ELANA technique with intraoperative and postoperative bypass patency were studied using logistic regression analysis. RESULTS: At the end of the operation, 146 (92%) of the 159 bypasses were patent. A first attempt to create a bypass was almost 8 times more likely (OR 7.6, 95% CI 2.1-27.5; p = 0.02) to result in a patent bypass than a second attempt. Administration of a small amount of heparin during the operation was also associated with bypass patency (OR 5.2, 95% CI 1.1-24.9; p = 0.04). One hundred twenty-three (77%) of the 159 bypasses were functional at patency assessments during the 1st month after the operation. Older age (OR 1.043 for every year of increase in age, 95% CI 1.010-1.076; p = 0.01), male sex (OR 2.9, 95% CI 1.3-6.5; p = 0.01), and high intraoperative bypass flow (OR 1.017 for every milliliter per minute increase in flow, 95% CI 1.004-1.030; p = 0.01) were associated with postoperative bypass patency. CONCLUSIONS: Attempts to create a second EC-IC ELANA bypass after the first one are more likely to fail, whereas administration of heparin to the patient during the procedure increases the intraoperative bypass patency rate. Postoperative patency results are better in male and in older patients. Intraoperative bypass flow measurements are essential because high bypass flow is an important determinant of postoperative patency. PMID- 19284229 TI - Minimal-access technique for distal catheter insertion during ventricular peritoneal shunt procedures: a review of 100 cases. AB - The authors report the safety and efficacy of using a percutaneous minimal-access insertion technique for distal shunt catheter placement in 100 cases. From June 2007 to March 2008, they attempted 100 minimal-access insertions of distal shunt catheters in 91 patients who required ventriculoperitoneal shunting. Using the minimal-access approach, they avoided utilizing laparoscopic assistance or a mini laparotomy in 91% of the cases. There were no bowel injuries or misplaced distal catheters. Additional outcomes in terms of operative times, cases that required conversion to open or laparoscopically assisted implantation, and infection rates are presented. They conclude that intraperitoneal shunt catheter placement can be safely and effectively accomplished using a simplified percutaneous minimal access insertion method that does not require direct laparoscopic visualization. PMID- 19284230 TI - Reduced limbic connections may contraindicate subgenual cingulate deep brain stimulation for intractable depression. AB - In this study, the authors performed deep brain stimulation (DBS) of the subgenual anterior cingulate cortex (SACC) in a patient with a history of bipolar disorder. After a right thalamic stroke, intractable depression without mood elevation or a mixed state developed in this patient. He underwent bilateral SACC DBS and died 16 months afterwards. Anatomical connections were studied in this patient preoperatively and postmortem using diffusion tractography (DT). A comparison of in vivo and high resolution ex vivo connectivity patterns was performed as a measure of the utility of in vivo DT in presurgical planning for DBS. Diagnostic measures included neuropsychological testing, preoperative and ex vivo DT, and macroscopic neuropathological assessment. Post-DBS depression rating scores did not improve. In vivo and ex vivo DT revealed markedly reduced limbic projections from the thalamus and SACC to the amygdala in the right (stroke affected) hemisphere. A highly selective right mediothalamic lesion was associated with the onset of refractory depression. Reduced amygdalar-thalamic and amygdalar-SACC connections could be a contraindication to DBS for depression. Correspondence between preoperative and higher resolution ex vivo DT supports the validity of DT as a presurgical planning tool for DBS. PMID- 19284231 TI - Entrapment of the glossopharyngeal nerve in patients with Eagle syndrome: surgical technique and outcomes in a series of 5 patients. AB - OBJECT: Eagle syndrome is characterized by unilateral pain in the oropharynx, face, and earlobe, and is caused by an elongated styloid process or ossification of the stylohyoid ligament with associated compression of the glossopharyngeal nerve. The pain syndrome may be successfully treated with surgical intervention that involves resection of the styloid process. Although nerve decompression is routinely considered a neurosurgical intervention, Eagle syndrome and its treatment are not sufficiently examined in the neurosurgical literature. METHODS: A review was performed of cases of Eagle syndrome treated in the Department of Neurosurgery at the University of Illinois at Chicago Medical Center over the last 7 years. The clinical characteristics, radiographic imaging, operative indications, procedural details, surgical morbidity, and clinical outcomes were collected and analyzed. RESULTS: Of the many patients with facial pain treated between 2001 and 2007, 7 were diagnosed with Eagle syndrome, and 5 of these patients underwent resection of the elongated styloid process. There were 4 women and 1 man, ranging in age from 20 to 68 years (mean 43 years). The average duration of disease was 11 years. In all patients, a preoperative workup revealed unilateral or bilateral elongation of the styloid process. All patients underwent resection of the styloid process on the symptomatic side using a lateral transcutaneous approach. There were no surgical complications. All patients experienced pain relief immediately after the operation. At the latest follow-up (average 46 months, range 7 months to 7.5 years) all but 1 patient maintained complete pain relief. In 1 patient, the pain recurred 12 months postoperatively and additional interventions were required. CONCLUSIONS: Eagle syndrome may be considered an entrapment syndrome of the glossopharyngeal nerve. It is a distinct clinical entity that should be considered when evaluating patients referred for glossopharyngeal neuralgia. The authors' experience indicates that patients with Eagle syndrome may be successfully treated using open resection of the elongated styloid process, which appears to be both safe and effective in terms of long lasting pain relief. PMID- 19284232 TI - Low-grade oligodendroglioma: an indolent but incurable disease? Clinical article. AB - OBJECT: The authors reviewed their institutional experience with pure low-grade oligodendroglioma (LGO), correlating outcomes with several variables of possible prognostic values. METHODS: Sixty-nine patients with WHO-classified LGOs were treated between 1992 and 2006 at the McGill University Health Center. Clinical, pathological, and radiological records were carefully reviewed. Demographic characteristics; the nature and duration of presenting symptoms; baseline neurological function; extent of resection; Karnofsky Performance Scale score; preoperative radiological findings including tumor size, location, and absence/presence of enhancement; and pathological data including chromosome arms 1p/19q codeletion and O-methylguanine-DNA methyltransferase promoter gene methylation status were all compiled. The timing and dose of radio- and/or chemotherapy, date of tumor progression, pathological finding at disease progression, treatment at time of disease progression, and status at the last follow-up were also recorded. RESULTS: The median follow-up period was 6.1 years (range 1.3-16.3 years). The majority (78%) of patients presented with seizures; contrast enhancement was initially seen in 16 patients (25%). All patients had undergone an initial surgical procedure: gross-total resection in 27%, partial resection in 59%, and biopsy only in the remaining 13%. Fifteen patients received adjuvant radiotherapy. Data on O-methylguanine-DNA methyltransferase promoter gene methylation status was available in 47 patients (68%) and in all but 1 patient for 1p/19q status. Survival at 5, 10, and 15 years was 83, 63, and 29%, respectively. Multivariate analysis showed that seizures at presentation and the absence of contrast enhancement were the only independent favorable prognostic factors for survival. The 5-, 10-, and 15-year progression-free survival rates were 46, 7.7, and 0%, respectively. CONCLUSIONS: This retrospective review confirms the indolent but progressively fatal nature of LGOs. Contrast enhancement was the most evident single prognostic factor. New treatment strategies are clearly needed in the management of this disease. PMID- 19284233 TI - Localization of bone marrow stromal cells to the injury site after intracerebral hemorrhage in rats. AB - OBJECT: Previous studies demonstrated that intravascular injection of bone marrow stromal cells (BMSCs) significantly improved neurological functional recovery in a rat model of intracerebral hemorrhage (ICH). To further investigate the fate of transplanted cells, we examined the effect of male rat BMSCs administered to female rats after ICH. METHODS: Twenty-seven female Wistar rats were subjected to ICH surgery. At 24 hours after ICH, these rats were randomly divided into 3 groups and injected intravenously with 1 ml phosphate-buffered saline or 0.5 million or 1 million male rat BMSCs in phosphate-buffered saline. To evaluate the neurological functional outcome, each rat was subjected to a series of behavioral tests (modified neurological severity score and corner turn test) at 1, 7, and 14 days after ICH. The rats were anesthetized intraperitoneally and killed, and the brain tissues were processed at Day 14 after ICH. Immunohistochemistry and in situ hybridization were used to identify cell-specific markers. RESULTS: The male rat BMSCs significantly improved the neurological functional outcome and also significantly diminished tissue loss when intravenously transplanted into the rats after ICH. Immunoassay for bromodeoxyuridine (BrdU) and neuronal markers demonstrated a significant increase in the number of BrdU-positive cells, which indicated endogenous neurogenesis, and a significant increase in the number of cells positive for immature neuronal markers. In situ hybridization showed that more BMSCs resided around the hematoma of the rats treated with the 1-million cell dose compared with the 0.5-million-cell-dose group. In addition, a subfraction of Y chromosome-positive cells were co-immunostained with the neuronal marker microtubule-associated protein-2 or the astrocytic marker glial fibrillary acidic protein. CONCLUSIONS: Male rat BMSCs improve neurological outcome and increase histochemical parameters of neurogenesis when administered to female rats after ICH. This study has shown that the intravenously administered male rat BMSCs enter the brain, migrate to the perihematomal area, and express parenchymal markers. PMID- 19284234 TI - Prediction of intracranial computed tomography findings in patients with minor head injury by using logistic regression. AB - OBJECT: The aim of this study was to develop a decision rule for physicians in developing countries to identify patients with minor head injury who will benefit from emergency brain CT scanning. METHODS: Three hundred eighteen patients with a history of blunt head trauma and a Glasgow Coma Scale (GCS) score >or= 13 who had presented within 12 hours of trauma underwent nonenhanced brain CT and were included in this prospective study. Computed tomography findings that necessitated neurosurgical care (either observation or intervention) were considered as positive findings. Logistic regression was used to develop the decision rule. RESULTS: Computed tomography scans were always normal in patients < 65 years old who did not have an obvious head wound, a raccoon sign, vomiting, memory deficit, or a decrease in their GCS score. Patients with 1 major criterion (GCS score < 14, raccoon sign, failure to remember the impact, age > 65 years, or vomiting) or 2 minor criteria (wound at the scalp or GCS score < 15) had an abnormal CT scan in 13% of the cases. CONCLUSIONS: The decision rule developed by the authors appears to be 100% sensitive and 46% specific for positive findings on brain CT and will, in developing countries, help clarify the decision to obtain scans. PMID- 19284235 TI - Intranasal delivery of erythropoietin plus insulin-like growth factor-I for acute neuroprotection in stroke. Laboratory investigation. AB - OBJECT: Individually, the cytokines erythropoietin (EPO) and insulin-like growth factor-I (IGF-I) have both been shown to reduce neuronal damage significantly in rodent models of cerebral ischemia. The authors have previously shown that EPO and IGF-I, when administered together, provide acute and prolonged neuroprotection in cerebrocortical cultures against N-methyl-D-aspartate-induced apoptosis. The aim of this study was to determine whether intranasally applied EPO plus IGF-I can provide acute neuroprotection in an animal stroke model and to show that intranasal administration is more efficient at delivering EPO plus IGF I to the brain when compared with intravenous, subcutaneous, or intraperitoneal administration. METHODS: The EPO and IGF-I were administered intranasally to mice that underwent transient middle cerebral artery occlusion (MCAO). Stroke volumes were measured after 1 hour of MCAO and 24 hours of reperfusion. To evaluate the long-term effects of this treatment, behavioral outcomes were assessed at 3, 30, 60, and 90 days following MCAO. Radiography and liquid scintillation were used to visualize and quantify the uptake of radiolabeled 125I-EPO and 125I-IGF-I into the mouse brain after intranasal, intravenous, subcutaneous, or intraperitoneal administration. RESULTS: Intranasal administration of EPO plus IGF-I reduced stroke volumes within 24 hours and improved neurological function in mice up to 90 days after MCAO. The 125I-EPO and 125I-IGF-I were found in the brain within 20 minutes after intranasal administration and accumulated within the injured areas of the brain. In addition, intranasal administration delivered significantly higher levels of the applied 125I-EPO and 125I-IGF-I to the brain compared with intravenous, subcutaneous, or intraperitoneal administration. CONCLUSIONS: The data demonstrate that intranasal EPO plus IGF-I penetrates into the brain more efficiently than other drug delivery methods and could potentially provide a fast and efficient treatment to prevent chronic effects of stroke. PMID- 19284236 TI - Efficacy of lamina terminalis fenestration in reducing shunt-dependent hydrocephalus following aneurysmal subarachnoid hemorrhage: a systematic review. Clinical article. AB - OBJECT: Chronic hydrocephalus requiring shunt placement is a common complication following aneurysmal subarachnoid hemorrhage (SAH). Controversy exists over whether microsurgical fenestration of the lamina terminalis during aneurysm surgery affords a reduction in the development of shunt-dependent hydrocephalus. To resolve this debate, the authors performed a systematic review and quantitative analysis of the literature to determine the efficacy of lamina terminalis fenestration in reducing aneurysmal SAH-associated shunt-dependent hydrocephalus. METHODS: A MEDLINE (1950-2007) database search was performed using the following keywords, singly and in combination: "ventriculoperitoneal shunt," "hydrocephalus," "subarachnoid hemorrhage," "aneurysm," "fenestration," and "lamina terminalis." Additional studies were manually singled out by scrutinizing references from identified manuscripts, major neurosurgical journals and texts, and personal files. A recent study from the authors' institution was also incorporated into the review. Data from included studies were analyzed using the chi-square analysis and Student t-test. The Cochran-Mantel-Haenszel test was used to compare overall incidence of shunt-dependent hydrocephalus. RESULTS: The literature search revealed 19 studies, but only 11 were included in this review, involving 1973 patients. The fenestrated and nonfenestrated cohorts (combined from the various studies) differed significantly with regard to patient sex, age, and clinical grade as well as aneurysm location (p=0.0065, 0.0028, 0.0003, and 0.017, respectively). The overall incidence of shunt-dependent hydrocephalus in the fenestrated cohort was 10%, as compared with 14% in the nonfenestrated cohort (p=0.089). The relative risk of shunt-dependent hydrocephalus in the fenestrated cohort was 0.88 (95% CI 0.62-1.24). CONCLUSIONS: This systematic review revealed no significant association between lamina terminalis fenestration and a reduced incidence of shunt-dependent hydrocephalus. The interpretation of these results, however, is restricted by unmatched cohort differences as well as other inherent study limitations. Although the overall literature supports lamina terminalis fenestration, a number of authors have questioned the technique's benefits, thus rendering its efficacy in reducing shunt-dependent hydrocephalus unclear. A well designed, multicenter, randomized controlled trial is needed to definitively address the efficacy of this microsurgical technique. PMID- 19284237 TI - Relevance of intracranial hypertension for cerebral metabolism in aneurysmal subarachnoid hemorrhage. Clinical article. AB - OBJECT: Intracranial hypertension, defined as intracranial pressure (ICP) >/= 20 mm Hg, is a complication typically associated with head injury. Its impact on cerebral metabolism, ICP therapy, and outcome has rarely been studied in patients with aneurysmal subarachnoid hemorrhage (aSAH); such an assessment is the authors' goal in the present study. METHODS: Cerebral metabolism was prospectively studied in 182 patients with aSAH. The database was retrospectively analyzed with respect to ICP. Patients were classified into 2 groups based on ICP. There were 164 with low ICP (<20 mm Hg) and 18 with high ICP (>or=20 mm Hg, measured>6 hours/day). Cerebral microdialysis parameters of energy metabolism, glycerol, and glutamate levels were analyzed hourly from the brain parenchyma of interest for 7 days. The 12-month outcome in these patients was evaluated. RESULTS: In the high ICP group, extended ICP therapy including decompressive craniectomy was necessary in 7 patients (39%). Cerebral glycerol levels and the lactate/pyruvate ratio were pathologically increased on Days 1-7 after aSAH (p<0.001). The excitotoxic neurotransmitter glutamate and glycerol, a marker of membrane degradation, further increased on Days 5-7, probably reflecting the development of secondary brain damage. An ICP>or=20 mm Hg was shown to have a significant influence on the 12-month Glasgow Outcome Scale (GOS) score (p=0.001) and was a strong predictor of mortality (OR=24.6; p<0.001). Glutamate (p=0.012), the lactate/pyruvate ratio as a marker of anaerobic metabolism (p=0.028), age (p<0.001), and Fisher grade (p=0.001) also influenced the GOS score at 12 months. CONCLUSIONS: The authors confirmed the relevance of intracranial hypertension as a severe complication in patients with aSAH. Because high ICP is associated with a severely deranged cerebral metabolism and poor outcome, future studies focusing on metabolism-guided, optimized ICP therapy could help minimize secondary brain damage and improve prognosis in patients with aSAH. PMID- 19284238 TI - Fenestration of the oculomotor nerve by a duplicated posterior cerebral artery and aneurysm. Case report. AB - The authors report a case of a posterior cerebral artery aneurysm arising from an aberrant arterial branch that was found to be splitting the oculomotor nerve. This 63-year-old man presented with headaches, and CT imaging revealed subarachnoid hemorrhage primarily within the basilar cisterns. Formal angiography and CT angiography revealed a saccular aneurysm arising from the posterior cerebral artery. Intraoperatively, the third cranial nerve was found to be fenestrated with the artery and aneurysm. PMID- 19284239 TI - History of the genesis of detachable coils. A review. AB - The genesis of detachable coils and the background of this genesis are described in this article. To frame the beginning developmental stages of the discovery of detachable coils, the previous extravascular-intravascular and endovascular techniques are presented, as well as the development of the various delivery systems. The experimental studies, initial clinical application, and crucial moment of the conception of detachable coils are also reported. PMID- 19284240 TI - The role of endoscopic third ventriculostomy in adult patients with hydrocephalus. AB - OBJECT: Endoscopic third ventriculostomy (ETV) is the treatment of choice for hydrocephalus, but the outcome is dependent on the cause of this disorder, and the procedure remains principally the preserve of pediatric neurosurgeons. The role of ETV in adult patients with hydrocephalus was therefore investigated. METHODS: One hundred ninety adult patients underwent ETV for hydrocephalus. Cases were defined as primary ETV (newly diagnosed, without a previously placed shunt) and secondary ETV (performed for shunt malfunctions due to infection or mechanical blockage). Causes of hydrocephalus included tumor, long-standing overt ventriculomegaly (LOVA), Chiari malformation Types I and II (CM-I and -II), aqueduct stenosis, spina bifida, and intraventricular hemorrhage (IVH). Successful ETV was defined as resolution of symptoms with shunt independence. Operative complications and ETV failure rate were investigated according to the causes of hydrocephalus and between the primary and secondary ETV groups. RESULTS: In the primary group, ETV was successful in 107 (83%) of 129 patients, including those with tumors (52 of 66), LOVA (21 of 24), CM-I (11 of 11 cases), CM-II (8 of 9), aqueduct stenosis (8 of 9), and IVH (2 of 2). In the secondary group, ETV was successful in 41 (67%) of 61 patients and was equally successful in cases of mechanical shunt malfunction (35 of 52 patients) and infected shunt malfunction (6 of 9 patients). The median time to ETV failure was 1.7 months in the primary group and 0.5 months in the secondary group. The majority of ETV failures occurred within the first 3 months, and thereafter, the Kaplan-Meier survival curves plateaued. There were no procedure-related deaths, and complications were seen in only 5.8% of cases. CONCLUSIONS: The success rate of ETVs in adults is comparable, if not better, than in children. In addition to the well-defined role of ETV in the treatment of hydrocephalus caused by tumors and aqueduct stenosis, ETV may also have a role in the management of CM-I, LOVA, persistent shunt infection, and IVH resistant to other CSF diversion procedures. PMID- 19284241 TI - The influence of focal brain cooling on neurophysiopathology: validation for clinical application. AB - OBJECT: Focal brain cooling has been recognized to have a suppressive effect on epileptiform discharges or a protective effect on brain tissue. However, the precise influence of brain cooling on normal brain function and histology has not yet been thoroughly investigated. The aim of this study was to investigate the neurophysiopathological consequences of focal cooling and to detect the threshold temperature that causes irreversible histological change and motor dysfunction. METHODS: The experiments were performed in adult male Sprague-Dawley rats (weighing 250-350 g) after induction of halothane anesthesia. A thermoelectric chip (6 x 6 x 2 mm) was used as a cooling device and was placed on the surface of the sensorimotor cortex after a 10 x 8-mm craniotomy. A thermocouple was placed between the chip and the brain surface. Focal cooling of the cortex was performed at the temperatures of 20, 15, 10, 5, 0, and -5 degrees C for 1 hour (5 rats in each group). Thereafter, the cranial window was repaired. Motor function was evaluated using the beam-walking scale (BWS) every day for 7 days. The rats were killed 7 days after the operation for histological examination with H & E, Kluver Barrera, glial fibrillary acidic protein, and terminal deoxynucleotidyl transferasemediated deoxyuridine triphosphate nick-end labeling stainings. The authors also euthanized some rats 24 hours after cooling and obtained brain sections by the same methods. RESULTS: The BWS score was decreased on the day after cooling only in the -5 degrees C group (p < 0.05), whereas the score did not change in the other temperature groups. Histologically, the appearance of cryoinjury such as necrosis, apoptosis, loss of neurons, and marked proliferation of astrocytes at the periphery of the lesion was observed only in the -5 degrees C group, while no apparent changes were observed in the other temperature groups. CONCLUSIONS: The present study confirmed that the focal cooling of the cortex for 1 hour above the temperature of 0 degrees C did not induce any irreversible histological change or motor dysfunction. These results suggest that focal brain cooling above 0 degrees C has the potential to be a minimally invasive and valuable modality for the treatment of severe brain injury or to assist in the examination of brain function. PMID- 19284242 TI - Balloon-assisted transarterial embolization of intracranial dural arteriovenous fistulas. AB - OBJECT: The authors report their preliminary experience using a balloon-assisted technique (BAT) in the transarterial embolization of intracranial dural arteriovenous fistulas (DAVFs). METHODS: The authors reviewed the prospectively collected data obtained in 7 consecutive patients with DAVFs in whom embolization was achieved using transarterially injected Onyx with either the venous or arterial BAT. Procedures were performed at the Division of Interventional Neuroradiology at the University of California at Los Angeles Medical Center between September 2005 and January 2008. RESULTS: Three patients presented with cortical venous reflux and 4 did not. Three patients underwent transarterial Onyx based embolization combined with transvenous balloon protection; the balloon was inflated in the transverse sinus in 2 of these patients and in the superior sagittal sinus in the third. One of them underwent an additional transarterial Onyx embolization with arterial BAT, whereas 4 other patients were treated with arterial BAT alone. The occipital artery was temporarily occluded with the balloon in 4 of these cases, whereas in the fifth, the authors used temporary balloon occlusion of the middle meningeal artery. Angiograms obtained immediately after embolization demonstrated complete or near-complete obliteration of the fistula in 6 patients and partial occlusion in 1 patient. There were no immediate or postprocedural complications. Two patients who presented with intracranial hemorrhage never suffered a second hemorrhage, and all other patients experienced either complete resolution or significant improvement of their symptoms. CONCLUSIONS: The BAT provides a new complementary method in the transarterial embolization of DAVFs that are not amenable to transvenous embolization. The venous BAT protects the patency of critical venous pathways, whereas the arterial BAT provides better control of the Onyx-based embolization. PMID- 19284243 TI - Distinct striatal targets in treating obsessive-compulsive disorder and major depression. AB - The ventral striatum, including the head of the caudate nucleus and the nucleus accumbens, is a putative target for deep brain stimulation (DBS) in the treatment of obsessive-compulsive disorder (OCD) and major depression (MD). However, the respective roles of these structures in the pathophysiology of OCD and MD remain to be clarified. To address this issue, DBS of the ventral striatum was tested in 2 patients with severely distressing and intractable forms of OCD and MD. Comparisons of clinical outcomes and anatomical data on electrode positioning showed that caudate nucleus stimulation preferentially alleviated OCD manifestations, whereas nucleus accumbens stimulation improved depressive symptoms. These findings suggest that the caudate nucleus and nucleus accumbens participate differently in the pathogenesis of both of these psychiatric conditions. PMID- 19284244 TI - Stereotactic coordinates associated with facial musculature contraction during high-frequency stimulation of the subthalamic nucleus. AB - OBJECT: High-frequency stimulation of the subthalamic nucleus (STN) in patients with parkinsonian symptoms is often used to ameliorate debilitating motor symptoms associated with this condition. However, individual variability in the shape and orientation of this relatively small nucleus results in multiple side effects related to the spread of electrical current to surrounding structures. Specifically, contraction of the muscles of facial expression is noted in a small percentage of patients, although the precise mechanism remains poorly understood. METHODS: Facial muscle contraction was triggered by high-frequency stimulation of 49 contacts in 18 patients undergoing deep brain stimulation of the STN. The mean coordinates of these individual contacts relative to the anterior commissure posterior commissure midpoint (also called the midcommissural point) were calculated to determine the location or structure(s) most often associated with facial contraction during physiological macrostimulation. RESULTS: The x, y, and z coordinates associated with contraction of the facial musculature were found to be 11.52, 1.29, and 1.15 mm lateral, posterior, and inferior to the midcommissural point, respectively. This location, along the lateral-anterior superior border of the STN, may allow for the spread of electrical current to the fields of Forel, zona incerta, and/or descending corticospinal/corticobulbar tracts. Because stimulation of corticobulbar tracts produces similar findings, these results are best explained by the spread of electrical current to nearby internal capsule axons coursing lateral to the STN. CONCLUSIONS: Thus, if intraoperative deep brain stimulation lead testing results in facial musculature contraction, placement of the electrode in a more medial, posterior position may reduce the amount of current spread to corticobulbar fibers and resolve this side effect. PMID- 19284245 TI - A prospective study of cerebral blood flow and cerebrovascular reactivity to acetazolamide in 162 patients with idiopathic normal-pressure hydrocephalus. AB - OBJECT: Cerebral blood flow (CBF) and cerebrovascular reactivity (CVR) to acetazolamide were investigated prospectively in 162 patients with a proposed diagnosis of idiopathic normal-pressure hydrocephalus (NPH). The aim of this study was to assess the usefulness of the measurement of CBF and CVR in determining which patients would be likely to benefit from shunt placement. METHODS: The mean CBF of the whole brain was measured according to the Patlak plot method by using technetium-99m hexamethylpropyleneamine oxime. The CVR value was obtained from the response to administration of 500 mg acetazolamide and calculated as the percentage change from the baseline mean CBF value. RESULTS: One hundred forty-six patients (90.1%) responded to shunt placement ("responders"), but 16 patients (9.9%) did not ("nonresponders"). No significant difference in preoperative CBF was observed between responders and nonresponders. Preoperative CVR was significantly impaired (p<0.0025) in responders compared with healthy controls, but not in nonresponders. Responders with the incomplete triad had a significant reduction (p<0.001) in preoperative CVR, but not in preoperative CBF, compared with healthy controls. Responders with the complete triad had significantly lower preoperative CBF and CVR than those with the incomplete triad (p<0.01 and p<0.05, respectively). Postoperative CBF and CVR increased significantly (p<0.025 and p<0.001, respectively) in responders. CONCLUSIONS: Both CBF and CVR decrease with the development of NPH, suggesting that hemodynamic ischemia may be responsible for manifestation of the symptoms. Impaired CVR and reduced CBF with the development of symptoms can be proposed as diagnostic criteria for idiopathic NPH. PMID- 19284246 TI - Kinetics and effect of integrin expression on human CD34(+) cells during murine leukemia virus-derived retroviral transduction with recombinant fibronectin for stem cell gene therapy. AB - The CH-296 recombinant fragment of human fibronectin is essential for murine leukemia virus (MLV)-derived retroviral transduction of CD34(+) cells for the purpose of stem cell gene therapy. Although the major effect of CH-296 is colocalization of the MLV-derived retrovirus and target cells at specific adhesion domains of CH-296 mediated by integrins expressed on CD34(+) cells, the precise roles of the integrins are unclear. We examined the kinetics of integrin expression on CD34(+) cells during the course of MLV-derived retrovirus-mediated gene transduction with CH-296. Flow cytometry revealed that the levels of both very late activation protein (VLA)-4 and VLA-5 on CD34(+) cells freshly isolated from cord blood were insufficient for effective MLV-derived retroviral transduction. However, increases were achieved during culture for preinduction and MLV-derived retrovirus-mediated gene transduction in the presence of a cocktail of cytokines. In addition, we confirmed by using specific antibodies that inhibition of the cell adhesion mediated by the integrins significantly reduced transduction efficiency, indicating that integrin expression is indeed important for CH-296-based MLV-derived retroviral transduction. Only a few cytokines are capable of inducing integrin expression, and stem cell factor plus thrombopoietin was found to be the minimal combination that was sufficient for effective transduction of an MLV-derived retrovirus based on CH-296. Our findings should be useful for improving the culture conditions for CH-296-based MLV derived retroviral transduction in stem cell gene therapy. PMID- 19284249 TI - Genius. PMID- 19284250 TI - Does exposure to palliative care prior to hospice make a difference for caregivers? PMID- 19284251 TI - The use of complementary and alternative medicine therapies by patients with advanced cancer and pain in a hospice setting: a multicentered, descriptive study. PMID- 19284252 TI - Type II oncologists. PMID- 19284253 TI - End-of-life traumatic stress disorder. PMID- 19284254 TI - Breaking bad news. PMID- 19284255 TI - Measure the social outcomes of nonprofits. PMID- 19284257 TI - Palliative care cost research can help other palliative care programs make their case. PMID- 19284258 TI - Center to advance palliative care inpatient unit operational metrics: consensus recommendations. AB - The need to standardize the prospective collection and analysis of data has been a cornerstone of education and technical assistance provided by the Center to Advance Palliative Care (CAPC). Data analysis is vital for strategic planning, quality improvement, and demonstration of program impact to hospital administrators, private funders, and policymakers. To develop a set of core measures, CAPC convened a consensus panel in 2008 to focus on operational metrics for hospital inpatient palliative care units. Operational metrics, as distinct from clinical, customer, and financial metrics, describe the characteristics of patients admitted to an inpatient unit, such as age, disease, location, length of stay, and disposition. The panel arrived at 11 metric domains, all but one of which can be used for either internal programmatic use or for external comparisons of service characteristics/impact between different hospitals. In an effort to ensure access to reliably high-quality palliative care data throughout the nation, hospitals are encouraged to collect and report outcomes for each of the 11 metric domains described here. PMID- 19284259 TI - The future of funding for palliative care research: suggestions for our field. PMID- 19284260 TI - Characteristics and correlates of dyspnea in patients with advanced cancer. AB - BACKGROUND: Dyspnea is a very distressing symptom present in the vast majority of patients with advanced cancer. There are limited data on the characteristics and correlates of dyspnea in this population. The purpose of this study was to characterize dyspnea, explore the differences between breakthrough and continuous presentations, and to determine factors associated with its intensity. METHODS: Prospective observational study among 70 patients with dyspnea referred to a palliative care service. Dyspnea was assessed using the Edmonton Symptom Assessment System (ESAS, 0-10) and the Oxygen Cost Diagram (OCD). Oximetry, pulmonary function tests, Hospital Anxiety and Depression Scale (HADS), and a detailed systematic evaluation of daily characteristics of dyspnea were performed. Other symptoms were recorded using the ESAS. RESULTS: Of 30 patients, 70 (43%) were female, median age was 58 (range, 28-87), and the most frequent cancer diagnosis were lung (31/70; 44%) and urologic (15/70; 21%). Constant dyspnea occurred in 27 of 70 (39%) patients, with 14 of 70 (20%) presenting breakthrough episodes. Breakthrough-only dyspnea occurred in 43 of 70 (61%). The majority of patients with breakthrough episodes (39/57; 68%) presented fewer than 5 episodes daily, most frequently lasting for less than 10 minutes (50/57; 88%). In univariate analyses ESAS dyspnea was associated with fatigue (p < 0.0001), sleep (p = 0.002), anxiety (p = 0.006), depression (p = 0.01), sensation of well being (p = 0.03), and with OCD (p = 0.001). In multivariate analysis, ESAS dyspnea was associated with fatigue (p = 0.001), forced expiratory volume (p = 0.004), pain (p = 0.01), and depression (p = 0.03). Dyspnea intensity significantly interfered with activities (general activity, p = 0.01, mood, p = 0.02, walking ability, p = 0.04, normal work p = 0.04, and enjoyment of life, p = 0.01). CONCLUSION: Dyspnea in patients with advanced cancer more frequently had breakthrough characteristics, was of very short duration, and interfered with daily activities. PMID- 19284261 TI - Medication kits for managing symptomatic emergencies in the home: a survey of common hospice practice. AB - INTRODUCTION: Alleviation of symptoms associated with advanced illness and dying is a fundamental goal and core principle of palliative care. Little research exists regarding hospice programs' practices for prescribing, dispensing, and utilizing medication kits in the home for management of uncontrolled symptoms. METHODS: We conducted a telephone survey of all 22 agencies in New Hampshire providing home hospice care. The survey inquired about the timing of medication kit ordering and availability, characteristics of prescribers and pharmacies, kit contents, costs, frequency of use, and perceived impact of kits. RESULTS: All programs' kits contained medications to treat pain and dyspnea, 81% for nausea and vomiting, and 76% for seizures. Eighty-six percent of agencies (18/21) reported that a medication within the kits was used in more than 50% of cases. Eighty-six percent reported the kits often averted hospital or emergency department visits. Oral, sublingual, and rectal routes of administration were common as was topical preparations of combination medications. Three programs included parenteral morphine in kits. Kits cost less than $50 for the majority of programs. CONCLUSION: Hospice programs commonly utilize kits containing prescription medications for the purpose of managing uncontrolled symptoms in the home. There is considerable variation in kit contents and practice. Programs believe that kits diminish emergency department visits and hospitalizations. Research is needed to more fully describe and study the outcomes of these practices. PMID- 19284262 TI - Investigating critically ill patients' and families' perceptions of likelihood of survival. AB - OBJECTIVE: This study investigated the perception of chance for survival among critically ill patients and surrogates and compared those perceptions to actual survival and to clinical estimates of illness severity. Secondary aims explored whether select demographic, clinical, or personal measures were associated with different perceptions of chance for survival. DESIGN: Prospective, sequential, observational, survey-based study. Primary measures were perception of chance for survival as compared to actual survival and Acute Physiology and Chronic Health Evaluation II (APACHE II) scores. SETTING: Tertiary care, academic medical intensive care unit (MICU). PATIENTS: Subjects were English-speaking adult MICU patients with a MICU length-of-stay greater than three days or their surrogates (n = 100). RESULTS: Respondents tended to be more optimistic regarding chance for survival than supported by actual survival (p = 0.07) or APACHE II tertile (p = 0.34). Secondary analyses found African American race, faith, or religion impacting health decision-making, and higher health status reports were associated with more optimistic perceptions of chance for survival. CONCLUSION: Patient/surrogate perceptions of chance for survival were not associated with either actual MICU survival or illness severity (APACHE II) highlighting an opportunity to better inform critically ill patients and families regarding prognosis. Clinician recognition of patients' and families' backgrounds and values might set the stage for such discussions. PMID- 19284263 TI - A conceptual analysis of spirituality at the end of life. AB - The definition of spirituality is the subject of endless debates in the empirical literature. This content analysis sought to: (1) exhaustively review the empirical literature on end-of-life spirituality to extract definitional elements of this concept and (2) elaborate on these definitional elements to create an integrative and inclusive definition of end-of-life spirituality based on the items retrieved. A search of the literature on spirituality published in the last 10 years was conducted via the the PsychINFO and MEDLINE databases. Seventy-one articles were selected based on specific inclusion criteria. A qualitative thematic analysis yielded 11 dimensions for the concept of end-of-life spirituality, namely: (1) meaning and purpose in life, (2) self-transcendence, (3) transcendence with a higher being, (4) feelings of communion and mutuality, (5) beliefs and faith, (6) hope, (7) attitude toward death, (8) appreciation of life, (9) reflection upon fundamental values, (10) the developmental nature of spirituality, and (11) its conscious aspect. The definition derived from this concept analysis, after being tested empirically, may be useful in informing the development of new measures of spirituality and new protocols to assess spirituality in clinical settings. PMID- 19284264 TI - Effects of red blood cell transfusion on anemia-related symptoms in patients with cancer. AB - The aim of this study was to assess the effects of red blood cell transfusion, and the subsequent increase in hemoglobin values, on anemia-related symptoms in a cohort of patients with cancer with different survival times. A red blood cell transfusion was recommended to a consecutive sample of patients with hemoglobin levels of 8 +/- 0.5 g/dL. The number of units to be ordered was decided according the hemoglobin values with a mean target of increasing the hemoglobin values by approximately 2 g/dL. Hemoglobin values, anemia-related signs and symptoms, including well-being, fatigue, and dyspnea, were recorded at admission (T0), 1 day after the last transfusion (T1), and 15 days afterward (T2) by telephone contact or visit. Well-being, fatigue, and dyspnea were measured on a numerical scale of 0-10. Sixty-one patients were recruited in the period of study. One hundred thirty-three units of red blood cells were transfused (mean 2.18, 95% confidence interval [CI] 0.6). Complete data were available for 40 patients. Hemoglobin values and well-being significantly increased after transfusion (T1), maintaining acceptable values 15 days afterward (T2). Significant changes in fatigue and dyspnea were found immediately after transfusion, although the effect was partially lost 15 days after transfusion. No statistical differences were found between patients with different survival times. Fatigue was significantly lower in patients with longer survival times in comparison with patients with shorter survival times (p = 0.04). Blood transfusion in patients with hemoglobin values of approximately 8 g/dL improved anemia-related symptoms on a short-term basis. This benefit is independent of the stage of disease and survival. However, the effects on dyspnea and fatigue tend to decrease within 15 days, despite the maintenance of hemoglobin values attained after transfusions, suggesting that other factors may play a role. PMID- 19284265 TI - Elder mistreatment reporting: differences in the threshold of reporting between hospice and palliative care professionals and adult protective service. AB - BACKGROUND: Underreporting of elder mistreatment by health professionals is a significant problem. PURPOSE: To investigate differences in elder mistreatment reporting threshold between hospice/palliative care (HPC) professionals and Adult Protective Services (APS); explore factors for not reporting elder mistreatment. DESIGN: Cross-sectional questionnaire. SETTING/PARTICIPANTS: Four HPC teams and 42 APS workers in one county. METHODS: Five hypothetical elder mistreatment case vignettes along a spectrum of severity were scored on Likert scales for likelihood of reporting or accepting the cases. HPC professionals were surveyed about their knowledge and beliefs about reporting elder mistreatment. RESULTS: All 42 APS workers and 74% of 73 (n = 54) HPC professionals completed the survey. In all but the most severe case of abuse, APS was more likely to accept reports of elder mistreatment than HPC professionals were in reporting (p < 0.002). HPC professionals had reported a mean of 2.5 (+/-2.8, standard deviation [SD]) cases in the last 5 years. Thirty percent of HPC professionals had suspected cases of elder mistreatment that was not reported in the last 5 years. The median difference between the total number of suspected and reported cases was 2 (+/ 4.6, SD). Eleven percent had ethical concerns about reporting and 63% were concerned about practical consequences of reporting. Only 37% correctly identified the reporting agencies that have jurisdiction over abuse that occurs in long-term care. Correct identification of long-term care reporting agencies correlated with whether the HPC professional had training in elder mistreatment (r = 0.35, p = 0.009). Although 96% would report physical abuse that they witnessed, only 63% would report abuse verbalized by the patient. CONCLUSION: There is evidence that significant differences exist in elder mistreatment reporting thresholds between APS and HPC professionals. This finding should encourage HPC professionals to discuss with APS the cases they are unsure about reporting. Future research is need on elder mistreatment in the HPC setting. PMID- 19284266 TI - Race does not influence do-not-resuscitate status or the number or timing of end of-life care discussions at a pediatric oncology referral center. AB - BACKGROUND: End-of-life care (EOLC) discussions and decisions are common in pediatric oncology. Interracial differences have been identified in adult EOLC preferences, but the relation of race to EOLC in pediatric oncology has not been reported. We assessed whether race (white, black) was associated with the frequency of do-not-resuscitate (DNR) orders, the number and timing of EOLC discussions, or the timing of EOLC decisions among patients treated at our institution who died. METHODS: We reviewed the records of 380 patients who died between July 1, 2001 and February 28, 2005. Chi(2) and Wilcoxon rank-sum tests were used to test the association of race with the number and timing of EOLC discussions, the number of DNR changes, the timing of EOLC decisions (i.e., DNR order, hospice referral), and the presence of a DNR order at the time of death. These analyses were limited to the 345 patients who self-identified as black or white. RESULTS: We found no association between race and DNR status at the time of death (p = 0.57), the proportion of patients with DNR order changes (p = 0.82), the median time from DNR order to death (p = 0.51), the time from first EOLC discussion to DNR order (p = 0.12), the time from first EOLC discussion to death (p = 0.33), the proportion of patients who enrolled in hospice (p = 0.64), the time from hospice enrollment to death (p = 0.2) or the number of EOLC discussions before a DNR decision (p = 0.48). CONCLUSION: When equal access to specialized pediatric cancer care is provided, race is not a significant factor in the presence or timing of a DNR order, enrollment in or timing of enrollment in hospice, or the number or timing of EOLC discussions before death. PMID- 19284267 TI - Study design, precision, and validity in observational studies. PMID- 19284268 TI - Can I stop the warfarin? A review of the risks and benefits of discontinuing anticoagulation. AB - Long-term anticoagulant therapy with warfarin is part of standard therapy for several disorders commonly present in patients seen in hospice and palliative care programs. Yet warfarin is also the drug most implicated in adverse drug reactions and its risks rise with increasing age and comorbidity. Clinicians caring for patients with multiple comorbidities or a limited life expectancy often are faced with the decision as to whether anticoagulation should be continued. Published guidelines for the use of warfarin in venous thromboembolism and nonvalvular atrial fibrillation are based on studies in which such patients were underrepresented or excluded. Our review of the randomized trials on which these guidelines are based shows that the annual risk of recurrent venous thromboembolism after stopping warfarin is 2%-10%. No similar evidence is available for patients with atrial fibrillation, but the published CHADS(2) index uses multiple factors to estimate stroke risk. The risk of bleeding complications with warfarin is most closely linked to degree of anticoagulation. Nutritional compromise and changes in drug therapy increase this risk and require that any patient remaining on warfarin must undergo frequent monitoring of anticoagulant effect. PMID- 19284269 TI - Neoplastic meningitis #135. PMID- 19284270 TI - Cardiopulmonary resuscitation survival in the hospital setting #179. PMID- 19284271 TI - A reflection on hospice. PMID- 19284272 TI - A third-year family medicine resident's first experience in palliative medicine. PMID- 19284273 TI - What about hospice, Dr. Gianakos? PMID- 19284274 TI - My friend, my patient. PMID- 19284275 TI - Immediate pain relief and improved structural stability after percutaneous vertebroplasty for a severely destructive vertebral compression fracture. AB - Approximately 10% to 30% of patients with cancer have metastases to the spine that require treatment. Spinal metastases can cause acute and chronic pain, compression fractures, spinal instability, and neurologic deficits. Vertebral compression fractures can be a significant cause of pain and impaired function. Radiation therapy is the standard treatment for patients with spinal metastases. However, this may not be the optimal treatment by itself for some patients when immediate pain relief is desirable. Percutaneous vertebroplasty is a mildly invasive treatment which involves the injection of cement, polymethylmethacrylate (PMMA) into a diseased vertebral body. We report a patient who presented with a severely destructive lesion and compression fracture at T9 with severe pain and impaired mobility. Percutaneous vertebroplasty provided immediate pain relief and restored structural stability of the diseased vertebra. PMID- 19284281 TI - JPM patient information. Understanding paracentesis. PMID- 19284284 TI - Differential prevalence of Plasmodium infections and cryptic Plasmodium knowlesi malaria in humans in Thailand. AB - BACKGROUND: A case of human infection with Plasmodium knowlesi has been recently discovered in Thailand. To investigate the prevalence of this malaria species, a molecular-based survey was performed. METHODS: Blood samples from 1874 patients were tested for Plasmodium species by microscopy and nested polymerase chain reaction. P. knowlesi was characterized by sequencing the merozoite surface protein 1 gene (msp-1). RESULTS: Of all Plasmodium species identified, P. falciparum, P. vivax, P. malariae, P. ovale, and P. knowlesi contributed 43.52%, 68.08%, 1.37%, 1.03%, and 0.57%, respectively. Mixed-species infections were more common in northwestern and southwestern regions bordering Myanmar (23%-24%) than in eastern and southern areas (3%-5%). In northwestern and southwestern regions, mixed-species infections had a significantly higher prevalence in dry than in rainy seasons (P < .001). P. knowlesi was found in 10 patients, mostly from southern and southwestern areas-9 were coinfected with either P. falciparum or P. vivax. Most of the P. knowlesi Thai isolates were more closely related to isolates from macaques than to isolates from Sarawak patients. The msp-1 sequences of isolates from the same area of endemicity differed and possessed novel sequences, indicating genetic polymorphism in P. knowlesi infecting humans. CONCLUSIONS: This survey highlights the widespread distribution of P. knowlesi in Thailand, albeit at low prevalence and mostly occurring as cryptic infections. PMID- 19284285 TI - Strong correlations of anti-viral capsid antigen antibody levels in first-degree relatives from families with Epstein-Barr virus-related lymphomas. AB - BACKGROUND: Markers of Epstein-Barr virus (EBV) infection include anti-viral capsid antigen (VCA) immunoglobulin (Ig) G. High anti-VCA titers are associated with EBV-related lymphoproliferation, such as Burkitt lymphoma (BL) and Hodgkin lymphoma (HL). METHODS: Intrafamilial correlations of anti-VCA IgG levels were studied in 3 settings: 127 families recruited through patients with HL in France (population A), 31 families recruited through patients with BL in Uganda (population B), and 74 large families from a general population in Cameroon (population C). Titers were determined by enzyme-linked immunosorbent assay (populations A and C) or by immunofluorescence analysis (population B). RESULTS: In populations A and B, the anti-VCA IgG titers of the relatives of patients with HL or BL increased significantly (P = .01 and P < .001, respectively) with those of the index case patient. In all 3 populations, anti-VCA IgG titers were significantly correlated (P < .001 for A, P = .002 for B, and P < .001 for C) between genetically related individuals (father-offspring, mother-offspring, and sibling-sibling) but not between spouses. Similar results were obtained for population A after adjustment for total IgG levels. In all cases, the pattern of correlations was consistent with a polygenic model, with heritability ranging from 0.32 to 0.48. CONCLUSION: These results provide evidence for the genetic control of anti-VCA IgG titers and pave the way for identification of the loci involved. PMID- 19284287 TI - Plasmodium knowlesi: finally being recognized. PMID- 19284286 TI - Early changes in hepatitis C virus (HCV) levels in response to peginterferon and ribavirin treatment in patients with chronic HCV genotype 1 infection. AB - Early changes in hepatitis C virus (HCV) RNA levels were assessed in a large cohort of African American and white patients with chronic hepatitis C due to HCV genotype 1 who underwent therapy with peginterferon alfa-2a and ribavirin in the Study of Viral Resistance to Antiviral Therapy of Hepatitis C (Virahep-C). Analyses were restricted to 341 patients who completed the first 28 days of therapy without dose modification. HCV RNA levels decreased in virtually all patients, but the amount of the change varied markedly. The overall 28-day decrease in HCV RNA levels was at least as predictive of a sustained virological response as the first- or second-phase viral kinetics responses. Factors associated with a smaller decrease in the HCV RNA level between baseline and day 28 included African American race, higher initial HCV RNA level, more severe hepatic fibrosis, and higher body weight. African American patients with similar 28-day decreases in viral levels as white patients were still less likely to achieve a sustained virological response. These results suggest that racial differences in the response to antiviral therapy are due to greater unresponsiveness to intracellular actions of interferon in African American individuals and that standard doses of peginterferon and ribavirin may be suboptimal for patients with higher body weights. Trial registration. ClinicalTrials.gov identifier: NCT00038974 . PMID- 19284288 TI - Racial differences in response to interferon-based antiviral therapy for hepatitis C virus infection: a hardwiring issue? PMID- 19284289 TI - O-glycosylation pattern of CD24 from mouse brain. AB - The cell adhesion molecule CD24 is a highly glycosylated glycoprotein that plays important roles in the central nervous system, the immune system and in tumor biology. Since CD24 comprises only a short protein core of approximately 30 amino acids and low conservation among species, it has been proposed that the functions of CD24 are mediated by its glycosylation pattern. Our present study provides evidence that interaction of CD24 with the cell adhesion molecule L1 is mediated by O-linked glycans carrying alpha2,3-linked sialic acid. Furthermore, de-N glycosylated CD24 was shown to promote or inhibit neurite outgrowth of cerebellar neurons or dorsal root ganglion neurons, respectively, to the same extent as untreated CD24. Therefore, this study is focused on the structural elucidation of the chemically released, permethylated CD24 O-glycans by electrospray ionization ion trap mass spectrometry. Our analyses revealed the occurrence of a diverse mixture of mucin-type and O-mannosyl glycans carrying, in part, functionally relevant epitopes, such as 3-linked sialic acid, disialyl motifs, Le(X), sialyl Le(X) or HNK-1 units. Hence, our data provide the basis for further studies on the contribution of carbohydrate determinants to CD24-mediated biological activities. PMID- 19284290 TI - Short double-stranded RNAs of specific sequence activate ribosomal TAK1-D and induce a global inhibition of translation. AB - We have previously shown that short double-stranded RNAs of specific sequence induce phosphorylation in the activation loop of splicing variant D of the transforming growth factor beta-activated protein kinase 1 (TAK1-D). Here, we further characterize this novel function of TAK1-D and the mechanisms of this dsRNA-triggered phenomenon. Using a dominant negative TAK1-D mutant we demonstrate that TAK1-D activation is functionally required to trigger the activation of p38 MAP kinase and c-JUN terminal kinase and to induce cell death in NCI-H460 cells. While total TAK1-D protein was found in the cytoplasm as well as in the ribosomal fraction, activated TAK1-D phosphorylated on T184 and T187 in the activation loop was found to be exclusively associated with the 80S ribosome. The association of TAK1-D with the ribosome suggests an involvement in translation-dependent signaling and we demonstrate here that dsRNA-mediated activation of TAK1-D leads to a downregulation of mRNA translation. In addition, we show that TAK1-D is also phosphorylated after the induction of ribotoxic stress. Our data indicate that TAK1-D plays a role in the signaling events triggered by selected types of ribotoxic stress. PMID- 19284291 TI - The solution structure of pGolemi, a high affinity Mena EVH1 binding miniature protein, suggests explanations for paralog-specific binding to Ena/VASP homology (EVH) 1 domains. AB - Ena/VASP homology 1 (EVH1) domains are polyproline binding domains that are present in a wide range of adaptor proteins, among them Ena/VASP proteins involved in actin remodeling and axonal guidance. The interaction of ActA, a transmembrane protein from the food-borne pathogen Listeria monocytogenes, with EVH1 domains has been shown to be crucial for recruitment of the host's actin skeleton and, as a consequence, for the infectivity of this bacterium. We present the structure of a synthetic high-affinity Mena EVH1 ligand, pGolemi, capable of paralog-specific binding, solved by NMR spectroscopy. This peptide shares the common pancreatic peptide fold with its scaffold, avian pancreatic peptide, but shows pivotal differences in the amino-terminus. The interplay of spatial fixation and flexibility appears to be the reason for its high affinity towards Mena EVH1. Combined with earlier investigations, our structural data shed light on the specificity determinants of pGolemi and the importance of additional binding epitopes around the residues Thr74 and Phe32 on EVH1 domains regulating paralog specificity. Our results are expected to facilitate the design of other high-affinity, paralog-specific EVH1 domain ligands, and serve as a fundament for the investigation of the molecular mode of action of EVH1 domains. PMID- 19284292 TI - Protein-specific glycosylation: signal patches and cis-controlling peptidic elements. AB - The term 'protein-specific glycosylation' refers to important functional implications of a subset of glycosylation types that are under direct control of recognition determinants on the protein. Examples of the latter are found in the formation of the mannose-6-phosphate receptor ligand on lysosomal hydrolases, and in polysialylation of NCAM, which are regulated via conformational signal patches on the protein. Distinct from these examples, the beta4-GalNAc modification of N linked glycans on a selected panel of proteins, such as carbonic anhydrase or glycodelin, was demonstrated recently to require specific protein (sequence) determinants proximal to the glycosylation site that function as cis-regulatory elements. Another example of such a cis-regulatory element was described for the control of mammalian O-mannosylation. In this case, the structural features of substrate sites within the mucin domain of alpha-dystroglycan are necessary, but not sufficient for determining the transfer of mannose to Ser/Thr. Evidence has been provided that an upstream-located peptide is also essential. Such cis controlling elements provide a higher level of protein specificity, because a putative glycosylation site cannot result from a single point mutation. Here, we highlight recent work on protein-specific glycosylation with particular emphasis on the above-cited examples and we will try to link protein-specific glycosylation to function. PMID- 19284293 TI - Release of endo-lysosomal cathepsins B, D, and L from IEC6 cells in a cell culture model mimicking intestinal manipulation. AB - IEC6 cells were used as an in vitro model system to study the effects of cell damage caused by mechanical manipulation of intestine epithelial cells. We constructed an apparatus that allowed analyzing the consequences of mechanical compression in a standardized and reproducible manner. Manipulation of IEC6 cells induced necrosis rather than apoptosis, and resulted in release of HMGB1, which is known to function as a trigger of inflammatory responses in vivo. Mechanical damage by traumatic injury of the intestine is accompanied by altered protease activities in the extracellular space, but only little is known about the possible contribution of endo-lysosomal cathepsins. Therefore, we tested the supernatants of manipulated cells in our in vitro model system for proteolytic activity and determined release rates by fluorimetric assays. Endo-lysosomal proteases, such as cathepsins B, D, and L, were released from damaged cells within the first 3 h after manipulation. While cathepsin L re-associated with the surfaces of neighboring cells, cathepsins B and D were present in the extracellular space as soluble enzymes. We conclude that our apparatus for mechanical manipulation can be used to approach surgical trauma, thereby focusing on epithelial cells of the intestine mucosa. PMID- 19284294 TI - Staphylococcal cysteine protease staphopain B (SspB) induces rapid engulfment of human neutrophils and monocytes by macrophages. AB - Abstract Circulating neutrophils and monocytes constitute the first line of antibacterial defence, which is responsible for the phagocytosis and killing of microorganisms. Previously, we have described that the staphylococcal cysteine proteinase staphopain B (SspB) cleaves CD11b on peripheral blood phagocytes, inducing the rapid development of features of atypical cell death in protease treated cells. Here, we report that exposure of phagocytes to SspB critically impairs their antibacterial functions. Specifically, SspB blocks phagocytosis of Staphylococcus aureus by both neutrophils and monocytes, represses their chemotactic activity and induces extensive, nonphlogistic clearance of SspB treated cells by macrophages. The proteinase also cleaves CD31, a major repulsion ('do not-eat-me') signal, on the surface of neutrophils. We suggest that both proteolytic degradation of repulsion signals and induction of 'eat-me' signals on the surface of leukocytes are responsible for the observed intensive phagocytosis of SspB-treated neutrophils by human monocyte-derived macrophages. Collectively, this may lead to the depletion of functional neutrophils at the site of infection, thus facilitating staphylococcal colonisation and spreading. PMID- 19284296 TI - Genotype distribution of estrogen receptor alpha polymorphisms in pregnant women from healthy and preeclampsia populations and its relation to blood pressure levels. AB - BACKGROUND: The estrogen receptor alpha gene (ESR1) seems to be a potential candidate in altering risk for preeclampsia due to important estrogenic biological effects. Despite its previously reported association with severe preeclampsia in combined genotype in one selected ethnic group, further studies in other populations are required. The objective of this study was to investigate the relationship of the gene polymorphisms of the estrogen receptor alpha with preeclampsia in a southwest Chinese population. METHODS: We genotyped two polymorphisms in the ESR1 gene (PvuII and XbaI) by polymerase chain reaction restriction fragment length polymorphism analysis in a Chinese population. RESULTS: In the case-control study which included 204 preeclampsia case subjects and 236 normal control subjects, the frequencies of allele T for the PvuII site and allele G for the XbaI site in the patients (41.67% and 23.53%, respectively) were similar to those observed in controls (43.01% and 19.70%, respectively). Furthermore, the combined genotype analysis of ESR1 PvuII and XbaI polymorphisms showed no increased risk of either severe or total preeclampsia in our study populations. However, it showed that the CC homozygotes and CT heterozygotes in control pregnant women had higher systolic blood pressure levels than TT homozygotes for the PvuII site after adjustment for age and body mass index (p<0.05). CONCLUSIONS: Our work does not provide evidence in favor of ESR1 PvuII and XbaI being associated with preeclampsia but with increased blood pressure levels in normal pregnant women, in a southwest Chinese population. PMID- 19284295 TI - Maternal visfatin concentration in normal pregnancy. AB - OBJECTIVE: Adipose tissue has now emerged as a powerful endocrine organ via the production of adipokines. Visfatin, a novel adipokine with diabetogenic and immuno-modulatory properties has been implicated in the pathophysiology of insulin resistance in patients with obesity and Type-2 diabetes mellitus. The aim of this study was to determine whether there are changes in the maternal plasma concentration of visfatin with advancing gestation and as a function of maternal weight. STUDY DESIGN: In this cross-sectional study, maternal plasma concentrations of visfatin were determined in normal weight and overweight/obese pregnant women in the following gestational age groups: 1) 11-14 weeks (n=52); 2) 19-26 weeks (n=68); 3) 27-34 weeks (n=93); and 4) >37 weeks (n=60). Visfatin concentrations were determined by ELISA. Non parametric statistics were used for analysis. RESULTS: 1) The median maternal plasma visfatin concentration was higher in pregnant women between 19-26 weeks of gestation than that of those between 11-14 weeks of gestation (P<0.01) and those between 27-34 weeks of gestation (P<0.01); 2) among normal weight pregnant women, the median plasma visfatin concentrations of women between 19-26 weeks of gestation was higher than that of those between 11-14 weeks (P<0.01) and those between 27-34 weeks (P<0.01); and 3) among overweight/obese patients, the median maternal visfatin concentration was similar between the different gestational age groups. CONCLUSION: The median maternal plasma concentration of visfatin peaks between 19 26 and has a nadir between 27-34 weeks of gestation. Normal and overweight/obese pregnant women differed in the pattern of changes in circulating visfatin concentrations as a function of gestational age. PMID- 19284297 TI - The selection and application of ssDNA aptamers against MPT64 protein in Mycobacterium tuberculosis. AB - BACKGROUND: Tuberculosis (TB) remains a major health problem affecting millions of people worldwide. One-third of the world's population is infected with Mycobacterium tuberculosis, the etiologic agent of TB. A simple and rapid method to diagnose TB is urgently needed to be developed. The procedure of systematic evolution of ligands by exponential enrichment (SELEX) is a method in which single-stranded oligonucleotides (called aptamers) are selected from a wide variety of sequences, based on their interaction with a target molecule. Aptamers have been used in numerous investigations as therapeutic or diagnostic tools. METHODS: In this study, we apply a SELEX method to develop aptamers against MPT64 protein from M. tuberculosis. On this basis, a sandwich assay scheme with the complex of aptamer-MPT64 was designed and tested the feasibility of detecting M. tuberculosis by detecting MPT64 protein levels in the culture filtrates of 77 samples including M. tuberculosis and other Mycobacterium species. RESULTS: There was a highly significant difference (p<0.01) between group A (non-TB Mycobacterium, bacille Calmette-Guerin) and group B (M. tuberculosis, M. bovis), when they were diagnosed with the sandwich assay scheme based on aptamer-protein complex to detect MPT64 protein levels in the culture filtrates of samples. When the cut-off point was at the optical density value of 0.58 (95%=0.764-0.946; Z=6.130, p=0.0001), the sandwich assay scheme based on aptamer-protein complex had a high sensitivity (negative ration, 24/27, 86.3%) and specificity (positive ration, 46/52, 88.5%). CONCLUSIONS: Aptamer of MPT64 as a new detection tool, to a certain extent, is feasible to diagnose Mycobacterium tuberculosis. PMID- 19284298 TI - Determination of nitrotyrosine concentrations in plasma samples of diabetes mellitus patients by four different immunoassays leads to contradictive results and disqualifies the majority of the tests. AB - BACKGROUND: In the course of type 2 diabetes mellitus, insulin resistance has a severe impact on endothelial function leading to decreased synthesis of nitric oxide (NO). Postprandial hyperglycemia leads to the generation of reactive oxygen species, which counteracts the beneficial NO effects. NO and superoxide combine very fast in solution to form peroxynitrite, which is a potent protein-oxidizing agent. The peroxynitrite concentrations can be indirectly monitored by the detection of nitrotyrosine residues in proteins, reflecting the extent of damage caused by oxidative stress. METHODS: Four commercially available nitrotyrosine specific immunoassays were evaluated by parallel measurement of nitrotyrosine in 224 serum samples derived from 16 patients with type 2 diabetes and 12 healthy controls (13 male and 15 female, age: 33+/-11 years) following a standardized meal. RESULTS: The available ELISA tests were not applicable for nitrotyrosine determination in human plasma samples due to technical issues and implausible results. However, a competitive luminescence assay was able to provide sufficient sensitivity and lead to clinically meaningful results in our test samples. CONCLUSIONS: All three ELISA methods were disqualified and conclusions previously derived from clinical experiments using these tests should be carefully reconsidered or reconfirmed. In the absence of a liquid tandem chromatography mass spectrometry reference method, the luminescence test appears to be the method of choice for determination of nitrotyrosine in human plasma. PMID- 19284299 TI - Translation termination and protein folding pathway genes are not correlated in gastric cancer. AB - BACKGROUND: The eukaryotic release factor 3 (eRF3) has been shown to affect both tubulin and actin cytoskeleton, suggesting a role in cytoskeleton assembly, mitotic spindle formation and chromosome segregation. Also, direct interactions between eRF3 and subunits of the cytosolic chaperonin CCT have been described. Moreover, both eRF3a and CCT subunits have been described to be up-regulated in cancer tissues. Our aim was to evaluate the hypothesis that eRF3 expression levels are correlated with the expression of genes encoding proteins involved in the tubulin folding pathways. METHODS: Relative expression levels of eRF1, eRF3a/GSPT1, PFDN4, CCT2, CCT4, and TBCA genes in tumour samples relative to their adjacent normal tissues were investigated using real time-polymerase chain reaction in 20 gastric cancer patients. RESULTS: The expression levels of eRF3a/GSPT1 were not correlated with the expression levels of the other genes studied. However, significant correlations were detected between the other genes, both within intestinal and diffuse type tumours. CONCLUSIONS: eRF3a/GSPT1 expression at the mRNA level is independent from both cell translation rates and from the expression of the genes involved in tubulin-folding pathways. The differences in the patterns of expression of the genes studied support the hypothesis of genetically independent pathways in the origin of intestinal and diffuse type gastric tumours. PMID- 19284306 TI - Automating the expansion process of human skeletal muscle myoblasts with suppression of myotube formation. AB - An intelligent culture system accompanied by automated operations (liquid transfer and cell passage) was newly developed to perform serial cultures of human skeletal muscle myoblasts. To realize a desired performance, a laminin coated surface was applied to myoblast expansion in a culture flask. It was found that the laminin coating enhanced the overall growth ability attributable not to shortening of the doubling time but to prevention of differentiation toward myotube formation, compared with that on a conventional plain surface. In addition, the effects of seeding density and confluence degree on the growth were investigated quantitatively in terms of cell attachment and division as well as proliferative cell population in the culture on the laminin-coated surface. With increasing in seeding density, the number of proliferative cells decreased at the end of culture accompanied by an increase in the confluence degree, which caused poor attachment of the passaged cells on the surface in the subsequent culture. The quantitative analyses of these cell behaviors helped us determine the appropriate seeding density and attainable confluence degree during one passage, which were 1.0 x 10(3) cells/cm(2) and 0.5 as the initial and boundary conditions, respectively. An automated culture system that could manage two serial cultures by monitoring the confluence degree was constructed. The automated operation with the intelligent determination of the time for passage was successfully performed without serious loss of growth activity, compared with manual operation using conventional flasks. These results indicated that the monitoring of confluence degree is effective to perform the culture passage of myoblasts, being contributable to automating the cell expansion process. PMID- 19284307 TI - Anti-MSP1 block 2 antibodies are effective at parasite killing in an allele specific manner by monocyte-mediated antibody-dependent cellular inhibition. AB - We investigated whether anti-merozoite surface protein-1 (MSP1) block 2 antibodies mediate the monocyte-dependent antibody-mediated cellular inhibition (ADCI) of Plasmodium falciparum. This study was performed because soluble molecules have been shown to trigger ADCI and because MSP1 block 2 is released following processing and is the target of cytophilic IgG3 responses in exposed populations. We assessed human anti-MSP1 block 2 antibodies against 4 P. falciparum strains that carry the 3 main block 2 sequence alleles. These antibodies were able to inhibit in vitro growth of P. falciparum only in cooperation with human monocytes, whereas no direct inhibition was observed. However, the ADCI effect was strictly allele specific. Our findings highlight a new mechanism involving MSP1 in the protection against malaria. PMID- 19284308 TI - Temperature sensitivity of cardiac function in pelagic fishes with different vertical mobilities: yellowfin tuna (Thunnus albacares), bigeye tuna (Thunnus obesus), mahimahi (Coryphaena hippurus), and swordfish (Xiphias gladius). AB - We measured the temperature sensitivity, adrenergic sensitivity, and dependence on sarcoplasmic reticulum (SR) Ca(2+) of ventricular muscle from pelagic fishes with different vertical mobility patterns: bigeye tuna (Thunnus obesus), yellowfin tuna (Thunnus albacares), and mahimahi (Coryphaena hippurus) and a single specimen from swordfish (Xiphias gladius). Ventricular muscle from the bigeye tuna and mahimahi exhibited a biphasic response to an acute decrease in temperature (from 26 degrees to 7 degrees C); twitch force and kinetic parameters initially increased and then declined. The magnitude of this response was larger in the bigeye tuna than in the mahimahi. Under steady state conditions at 26 degrees C, inhibition of SR Ca(2+) release and reuptake with ryanodine and thapsigargin decreased twitch force and kinetic parameters, respectively, in the bigeye tuna only. However, the initial inotropy associated with decreasing temperature was abolished by SR inhibition in both the bigeye tuna and the mahimahi. Application of adrenaline completely reversed the effects of ryanodine and thapsigargin, but this effect was diminished at cold temperatures. In the yellowfin tuna, temperature and SR inhibition had minor effects on twitch force and kinetics, while adrenaline significantly increased these parameters. Limited data suggest that swordfish ventricular muscle responds to acute temperature reduction, SR inhibition, and adrenergic stimulation in a manner similar to that of bigeye tuna ventricular muscle. In aggregate, our results show that the temperature sensitivity, SR dependence, and adrenergic sensitivity of pelagic fish hearts are species specific and that these differences reflect species specific vertical mobility patterns. PMID- 19284309 TI - siRNA-mediated integrin-linked kinase suppression: nonspecific effects of siRNA/cationic liposome complexes trigger changes in the expression of phosphorylated-AKT and mTOR independently of ILK silencing. AB - Short interfering RNA targeting ILK (ILK siRNA) could be used to treat patients with cancers where constitutive activation of the AKT/PI3K pathway is prominent (e.g., those cancers lack functional PTEN). It is generally believed that siRNA therapeutics will require the use of delivery systems and lipid-based formulations containing cationic lipids (CLs) are a viable option. However, CLs are known to be toxic and exposure to CLs can influence cell survival pathways. This study characterized how CLs combine with ILK siRNA to influence the AKT/PI3K pathway. Using PTEN-negative cell lines (PC3 castration-insensitive prostate cancer cells and U251 glioma cancer cells), the influence of CLs on the downstream consequences of ILK silencing was determined. When comparing nucleofection (an electroporation method that does not require the use of CLs) and CLs as means to deliver ILK siRNA, a 12- to 30-fold increase in siRNA delivery was achieved when using a CL formulation, yet ILK suppression was less efficient. Importantly, time-dependent signaling consequences associated with ILK silencing, including suppression of phosphorylated (serine 473)-AKT and changes in mTOR expression, were observed independently of ILK suppression when the target cells were exposed to cationic lipids following nucleofection-based delivery of ILK siRNA. PMID- 19284310 TI - Antimetastatic potential of PAI-1-specific RNA aptamers. AB - The serine protease inhibitor plasminogen activator inhibitor-1 (PAI-1) is increased in several cancers, including breast, where it is associated with a poor outcome. Metastatic breast cancer has a dismal prognosis, as evidenced by treatment goals that are no longer curative but are largely palliative in nature. PAI-1 competes with integrins and the urokinase plasminogen activator receptor on the surface of breast cancer cells for binding to vitronectin. This results in the detachment of tumor cells from the extracellular matrix, which is critical to the metastatic process. For this reason, we sought to isolate RNA aptamers that disrupt the interaction between PAI-1 and vitronectin. Through utilization of combinatorial chemistry techniques, aptamers have been selected that bind to PAI 1 with high affinity and specificity. We identified two aptamers, WT-15 and SM 20, that disrupt the interactions between PAI-1 and heparin, as well as PAI-1 and vitronectin, without affecting the antiprotease activity of PAI-1. Furthermore, SM-20 prevented the detachment of breast cancer cells (MDA-MB-231) from vitronectin in the presence of PAI-1, resulting in an increase in cellular adhesion. Therefore, the PAI-1 aptamer SM-20 demonstrates therapeutic potential as an antimetastatic agent and could possibly be used as an adjuvant to traditional chemotherapy for breast cancer. PMID- 19284311 TI - Stem-loop RT-PCR quantification of siRNAs in vitro and in vivo. AB - RNA interference (RNAi) is a mechanism in which the introduction of small interfering RNAs (siRNAs) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. Applications of RNAi include gene function and pathway analysis, target identification and validation, and therapeutics. There is a need to develop reliable and easy-to-use assays to evaluate siRNA delivery efficiency and distribution, study pathways, and stability of siRNAs in cells (posttransfection) and in animals (postinjection). We have leveraged the Applied Biosystems TaqMan-based stem-loop RT-PCR technology, originally developed for quantification of endogenous microRNAs in cells, to fulfill these needs. The application protocols developed enable robust quantification of siRNA, including chemically modified siRNA molecules, in vitro and in vivo. PMID- 19284312 TI - Impact of obesity on the left ventricular functions and morphology of healthy Asian Indians. AB - BACKGROUND: The aim of this study was to determine the impact of obesity on alteration of left ventricular (LV) functions and morphology in nondiabetic, nonhypertensive, and normo-lipidemic obese Asian Indians. A total of 239 consecutive Asian Indians (175 males and 64 females, ages 17-64 years) were divided into obese and nonobese groups based on body mass index (BMI), waist-to hip circumference ratio (W-HR), and percentage of body fat (%BF). METHODS: Anthropometry (BMI, W-HR), %BF, and two-dimensional echocardiography including tissue Doppler imaging (TDI) were performed for all the subjects. The unpaired t test was applied after matching age and gender in all the comparison groups. Nonobese subjects acted as controls for the obese subjects (cases). RESULTS: Obese subjects had a larger LV end-diastolic diameter (P < 0.001), LV end systolic diameter (P < 0.001), and LV mass (P < 0.001) as compared to the nonobese subjects. Subclinical systolic dysfunction was apparent in obese subjects only on TDI in the form of reduced systolic mitral annular velocity (P = 0.009). Diastolic dysfunction, as suggested by a lower ratio of early to late transmitral ventricular filling velocity (E/A), lower early to late (Em/Am) diastolic mitral annular velocity, and a higher E/Em ratio (P < 0.001, p = 0.001 and P < 0.001, respectively), was noted in the obese cohort. In addition, the left atrial diameter (P < 0.001) was also increased in obese subjects. Alteration of LV morphology and function correlated with the anthropometric variables BMI, W HR, and %BF. CONCLUSIONS: Asian Indians with uncomplicated obesity (without associated co-morbidities) had significant morphological and functional cardiac dysfunction (systolic and diastolic), which correlated with anthropometric variables. PMID- 19284313 TI - Obesity is not a risk factor in children with reflux esophagitis: a retrospective analysis of 738 children. AB - Obesity has been associated with various gastrointestinal diseases in children, but the role of obesity in gastroesophageal reflux disease (GERD) has not been clearly established. The aim of the study was to investigate whether obesity and/or being overweight are risk factors for reflux esophagitis in children. A retrospective analysis of endoscopy charts was reviewed. Demographic, weight, height, and histology results were obtained from each patient. The body mass index (BMI) and BMI Z-score were calculated according to known formula. The diagnosis of GERD was established by histology. The charts of 738 children were reviewed; of these, 345 (47%) children were overweight or obese. Histological findings compatible with GERD were found in 254 (65%) children with normal weight, 111 (69%) overweight children, and 126 (68%) obese children (P > 0.05). Among those reviewed, the mean age of children with normal weight was significantly younger than that of overweight or obese children (P = 0.0001). A single variant analysis showed a significant association between GERD and male gender (P = 0.0001). Multivariant analysis (gender, age, and BMI Z-score) showed that GERD was significantly associated with male gender (P < 0.0001), but not with age (P = 0.443) or BMI Z-score (P = 0.098). In symptomatic children with histologically proven GERD, only male gender was an independent risk factor for GERD, not obesity or being overweight. Large, prospective studies in children that capture a larger spectrum of GERD are clearly warranted. PMID- 19284314 TI - Metabolic syndrome and cancer. AB - Since its first description by Reavan in 1988, accepted criteria for clinical identification of the components of metabolic syndrome have been promulgated by the National Cholesterol Education Program/Adult Treatment Panel III (NCEP/ATP III) and the World Health Organization (WHO) as well as the International Diabetes Federation (IDF), and the American Association of Clinical Endocrinologists (AACE). Insulin resistance is a common metabolic abnormality underlying type 2 diabetes mellitus and is also an independent risk factor for cardiovascular disease. Although ATP III identified cardiovascular disease (CVD) as the primary clinical outcome of the metabolic syndrome, we now have evidence that metabolic syndrome is associated with type 2 diabetes mellitus, polycystic ovarian disease, nonalcoholic fatty liver disease, and possibly some cancers. This review summarizes evidence in support of the relationship between metabolic syndrome and various cancers and possible underlying mechanisms and therapeutic interventions. PMID- 19284315 TI - Is a unified definition of metabolic syndrome needed? Comparison of three definitions of metabolic syndrome in 60-year-old men and women. AB - BACKGROUND: There are three commonly used definitions of the metabolic syndrome, making scientific studies hard to compare. The aim of this study was to investigate agreement in the prevalence of the metabolic syndrome defined by three different definitions and to analyze definition and gender differences. METHODS: A population-based, cross-sectional study of a total of 4232 participants--2039 men and 2193 women, aged 60 years--was employed. Three different metabolic syndrome definitions were compared: European Group for the Study of Insulin Resistance (EGIR), International Diabetes Federation (IDF), and National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III). Medical history, socioeconomic information, and lifestyle data were collected by a questionnaire. A medical examination including laboratory tests was performed. Significant factors for the metabolic syndrome were calculated by multivariate logistic regression. RESULTS: Forty five percent of men and 30% of women met the criteria for the metabolic syndrome by any definition, but only 17% of men and 9% of women met the criteria of all three definitions. The highest agreement was found between IDF and NCEP ATP III definition. Two significant associations were identified in both men and women by the three metabolic syndrome definitions; former smokers were highly associated with the metabolic syndrome (odds ratio [OR] congruent with 1.5), and regular physical activity (OR congruent with 0.6) was inversely associated with the metabolic syndrome. CONCLUSIONS: Depending on the definition used, different individuals were identified as having the metabolic syndrome, which affects the reliability of interpretations to be made from scientific studies of the metabolic syndrome. Unified criteria are warranted. Physicians facing a physically inactive former smoker may consider diagnosing metabolic syndrome. PMID- 19284316 TI - Correlation of a [13C]glucose breath test with surrogate markers of insulin resistance in urban and rural Asian Indians. AB - BACKGROUND: High rates of insulin resistance (IR), which contribute to a high prevalence of cardiovascular disease (CVD), have been noted in Asian Indians. Rapid, effective measures of IR could identify individuals at risk for CVD in this population. A [(13)C]glucose breath test has been shown to correlate significantly with invasive measures of IR in a Caucasian cohort. We hypothesized that the breath test would correlate significantly with surrogate measures of IR in Asian Indians. METHODS: A total of 49 urban and 49 rural Asian Indian subjects underwent the (13)C breath test and 2-hour oral glucose tolerance testing. Correlations were performed between the breath test results and surrogate measures of IR including the homeostasis model of insulin resistance (HOMA). These two indices were also correlated with body mass index (BMI). RESULTS: In the overall cohort, the breath test correlated significantly with HOMA (r = 0.40; P < 0.0001), waist circumference (WC) (r = -0.70; P < 0.0001), and BMI (r = -0.59; P < 0.0001). The breath test correlated significantly with BMI in the urban and rural cohorts (r = -0.65; P < 0.0001 and r = -0.36; P = 0.01) and with HOMA (r = -0.55; P = 0.0001) in the urban cohort. There was no significant correlation between the breath test and HOMA (r = -0.07; P = 0.61) in the rural cohort. When corrected for WC and BMI, the correlation between the breath test and HOMA in the urban cohort was no longer significant (r = -0.08; P = 0.57). CONCLUSIONS: The (13)C breath test correlated significantly with HOMA values in urban but not in more insulin-sensitive rural subjects and yielded no incremental information over BMI. Further refinement of the [(13)C]glucose breath test is necessary prior to its use as a screening test for IR in Asian Indians. PMID- 19284317 TI - Factor analysis of the individual components of the metabolic syndrome among elderly identifies two factors with different survival patterns--a population based study. AB - BACKGROUND: Factor analysis reduces a set of directly measured variables into a smaller set of underlying factors representing unique statistically independent domains termed "factors." In middle-aged people, previous factor analyses of the components of the metabolic syndrome have identified two to four factors. Many of these analyses report blood pressure as one of these factors. METHODS: We performed a factor analysis of the individual continuous components of the metabolic syndrome among 198 men and 203 women comprising 65% of all 75 year olds from the city of Vasteras in Sweden. RESULTS: The metabolic syndrome comprised two factors. Factor 1, the metabolic factor, consisted of waist circumference, high-density lipoprotein cholesterol, triglycerides, and fasting glucose, whereas factor 2, the blood pressure factor, consisted of systolic and diastolic blood pressure. These two factors explained 58% (factor 1, 31%, and factor 2, 27%) of the total variation in men. The corresponding figures for women were 63% (factor 1, 36%, and factor 2, 27%). Factor 1 was significantly related to a decreased 10 years of survival for men, with a hazard ratio (HR) of 1.30 (95% confidence interval [CI] 1.07-1.58, P = 0.008), and nearly significantly for women, with a HR of 1.27 (95% CI 0.98-1.64, P = 0.071). In a pooled analysis adjusting for sex, known cardiovascular disease, and hypertension, high blood pressure, and current smoking, the HR for factor 1 was 1.20 (95% CI 1.02-1.49, P = 0.026). Factor 2 was not significantly related to survival. CONCLUSIONS: Factor analysis of the basic variables of the metabolic syndrome among 75 year olds from a general population identified a metabolic factor and a blood pressure factor. The former factor was significantly related to 10-year survival and the relationship remained after adjusting for known cardiovascular disease and hypertension and current smoking. PMID- 19284318 TI - Association of leukocyte count with varying degrees of glucose intolerance in Asian Indians: the Chennai Urban Rural Epidemiology Study (CURES-26). AB - OBJECTIVE: This study assessed the association of leukocyte count with different grades of glucose intolerance in Asian Indian subjects. METHODS: Three groups of subjects were recruited from the Chennai Urban Rural Epidemiology Study (CURES), a population-based study, representative of Chennai (formerly Madras), a city in southern India. Group 1 represented normal glucose tolerance (NGT) (n = 840), group 2 included impaired glucose tolerance (IGT) (n = 180), and group 3 included type 2 diabetes (n = 1170). Anthropometric measurements including weight, height, and waist measurements were obtained using standardized techniques. Leukocyte count was measured by an automated flow cytometry instrument (Sysmex SF-3000, Japan). Fasting insulin was measured by enzyme-linked immunosorbent assay and insulin resistance was calculated using the homeostasis model assessment (HOMA IR). RESULTS: Subjects with diabetes (8.0 +/- 1.5 x 10(3)/microL) and IGT (7.9 +/ 1.3 x 10(3)/microL) had a significantly higher mean leukocyte count compared to the NGT group (7.4 +/- 1.5 x 10(3)/microL) (P < 0.001). Leukocyte count was significantly increased in NGT subjects with insulin resistance (IR) as measured by HOMA-IR (7.5 +/- 1.5 x 10(3)/microL; P < 0.001) compared to NGT subjects without IR (7.0 +/- 1.4 x 10(3)/microL). Regression analysis showed that there was a linear increase in mean leukocyte count with increasing severity of glucose intolerance, even after adjusting for age, waist circumference, and HOMA-IR. CONCLUSIONS: Among Asian Indians who are known to have high risk of premature coronary artery disease and diabetes, a significant association exists between leukocyte count and glucose intolerance. PMID- 19284319 TI - Cytochrome oxidase 2D6 gene polymorphism in primary open-angle glaucoma with various effects to ophthalmic timolol. AB - AIMS: Timolol is used topically for the treatment of glaucoma and metabolized by cytochrome P450 (CYP) 2D6 in the liver. The aim of this study is to test the hypothesis that CYP 2D6 single-nucleotide polymorphism (SNP) is associated with drug effects of ophthalmic timolol. METHODS: A total of 133 primary open-angle glaucoma (POAG) subjects underwent the ophthalmic single timolol administration and the drug effects were observed, including lowering the effects of intraocular pressure (IOP) and side effects (i.e., appearing bradycardia). Eight SNPs of CYP2D6 were investigated in 73 subjects by a SNPstream genotyping system. The relationship between the effects of timolol and CYP2D6 Arg296Cys and Ser486Thr genotype distribution in these POAG subjects was analyzed. RESULTS: Topical timolol administration had significant effect on IOP (P = 0.000) and heart rate (HR) (P = 0.000) in all 133 subjects, and individual ocular hypotensive effect of timolol varied between 0 and 23 mmHg. Individual effect of HR varied between -31 and 10 beats per minute, in the present study. According to SNP genotyping in 73 subjects, there was no significant difference of IOP between subjects with different CYP2D6 Arg296Cys (P = 0.308) or Ser486Thr genotypes (P = 0.741). The effect of timolol on HR was significantly different between subjects with different Arg296Cys genotypes (P = 0.046). Timolol-induced bradycardia tended to occur in subjects with Arg296Cys CT and TT genotype when compared with CC genotype (P = 0.009). CONCLUSIONS: CYP2D6 SNP Arg296Cys appeared to be correlative with the intersubject variability seen with timolol in POAG subjects. Subjects with CC genotype trended to avoid timolol-induced bradycardia, and subjects with TT genotype trended to have poorer timolol-induced ocular hypotensive effects. PMID- 19284320 TI - Development of a topical polymeric mucoadhesive ocular delivery system for azithromycin. AB - PURPOSE: Azithromycin is an azalide class of antibiotic with pharmacodynamics that have made it a valuable agent in the treatment of soft tissue infections. In ophthalmology, oral administration of azithromycin has been proven effective for the treatment of trachoma. However, topical formulations of azithromycin to treat ocular surface infections have been challenging to develop because of the drug's hydrophobicity and instability in aqueous solutions at pH levels that are comfortable in the eye. The design of a polycarbophil polymer-based delivery system for a topical formulation of azithromycin was evaluated for its ability to provide drug stability, comfort, and increased retention of the formulation in the eye. METHODS: Formulations of 0.5% and 1.0% azithromycin were created in polycarbophil, a lightly cross-linked polyacrylic acid polymer that was adjusted to a viscosity, pH, and osmolality that are suitable for dispensing in the eye. RESULTS: The polycarbophil-based ophthalmic delivery system, DuraSite (InSite Vision, Alameda, CA), helps solubilize azithromycin and retard its degradation in aqueous solution. The formulation was stable at room temperature as well as 5 degrees C. Upon administration of a single drop of 1% azithromycin in DuraSite ophthalmic solution in rabbits' eyes, tear concentrations of azithromycin ranged from 87 to 288 microg/g and high concentrations were sustained for over a 24-h period. CONCLUSIONS: Azithromycin can be developed as an eyedrop in an aqueous ocular delivery system for the treatment of ocular surface infections. The ocular delivery system, DuraSite solubilizes azithromycin at a high concentration in an aqueous solution and protects it from degradation during manufacture and storage. The development of azithromycin in this delivery system enhances the antibiotic's usefulness in ophthalmology for the topical treatment of ocular surface bacterial infections and lid margin diseases. PMID- 19284321 TI - The impact of ocular adverse effects in patients treated with topical prostaglandin analogs: changes in prescription patterns and patient persistence. AB - PURPOSE: To document patient/physician perceptions of adverse effects and their relationship to medication changes among patients prescribed prostaglandin analogs. METHODS: Medical/pharmacy claims (private U.S. health network) identified patients filling initial topical ocular hypotensive prescriptions from 2001 to 2004; 300 open-angle glaucoma patients prescribed a prostaglandin analog and 103 ophthalmologists were selected by algorithm for telephone interviews. Medical charts for 225/300 interviewed and 75 non-interviewed patients were abstracted. Medication patterns were assessed in pharmacy claims data. Frequency of adverse effects noted by physicians and associations with medication change decisions were examined in charted data. Patients' experiences with adverse effects were compiled from surveys. RESULTS: In patients treated with latanoprost (N = 4,071), bimatoprost (N = 1,199), or travoprost (N = 1,001), continuous refill of medication through 1 year was seen in 11%, 9%, and 5% of patients, respectively (P = 0.0001; retrospective pharmacy claims). Adverse effects were the second most common reasons noted by physicians for switching medications after lack of efficacy (19% vs. 43%, respectively). Adverse effects were noted in 65% of patient charts. Hyperemia was the most common adverse effect occurring with at least one other adverse effect in 48% of patients with the condition. CONCLUSIONS: Ocular adverse effects, particularly hyperemia, negatively affect patient continuation with therapy and switching. PMID- 19284322 TI - Acetazolamide influences ocular pulse amplitude. AB - PURPOSE: To investigate the effect of systemic acetazolamide on ocular pulse amplitude (OPA) and on intraocular pressure (IOP). METHODS: We determined OPA, IOP, mean arterial blood pressure and heart rate (HR) in 17 patients suffering from hypertensive primary open-angle glaucoma (htPOAG) and 22 healthy controls. Measurements were taken before and 2 h after oral intake of 500 mg of acetazolamide. RESULTS: Mean OPA decreased 20.24% (SEM 2.95%) from 2.76 mmHg (SEM 0.16 mmHg) to 2.14 mmHg (SEM 0.13 mmHg) 2 h after oral administration of 500-mg acetazolamide. IOP dropped by 2.98 mmHg (SEM 0.28 mmHg), corresponding to a 17.19% (SEM 1.56%) reduction. The decrease in IOP correlated with the decrease of OPA (r(2) = 0.4, P < 0.0001). CONCLUSIONS: Ocular pulse amplitude measurements are influenced by oral administration of acetazolamide. There was a decrease in the pulsatile component of choroidal blood flow and in IOP in both htPOAG and healthy controls. This needs to be taken into account for measurement of OPA with dynamic contour tonometry. PMID- 19284323 TI - Complications and safety profile of posterior subtenon injection of triamcinolone acetonide. AB - PURPOSE: Posterior subtenon injection (PSTI) of triamcinolone acetonide (TA) is one of the delivery methods for the treatment of posterior uveitis, diabetic macular edema, and cystoid macular edema secondary to retinal vein occlusions. The aim of this investigation was to assess the rates of various complications associated with PSTI and to determine which factors are associated with the development of complications. METHODS: This investigation was a retrospective interventional case series, and one retinal specialist from 2005 to 2007 conducted all of the procedures. Patients were followed up for a mean time of 11.9 +/- 5.4 months after receiving 40 mg (1.0 mL) of nonfiltered TA and all complications associated with the injection procedure or with TA itself were noted. RESULTS: A total of 159 eyes in 158 patients were treated by PSTI of TA. Of 159 eyes, 145 eyes (91.2%) were injected once, 13 eyes (8.2%) were injected twice, and one eye (0.6%) was injected three times. The mean age of the 158 patients was 61.6 +/- 9.5 years. Eighteen eyes (11.3%) required glaucoma medication due to increased intraocular pressure (IOP) after injection. In addition, cataracts in three eyes (2.1%) progressed with biomicroscopic findings, as compared with the fellow eyes. There were no cases of mechanical injury, infection or ulceration of any part of eye, and eyelid complications. A conjunctival cyst at the injection site developed in one eye. CONCLUSIONS: In this study, PSTI of TA resulted in no severe complications such as endophthalmitis or retinal detachment, and a relatively less risk of IOP complications and cataract progression. PSTI of TA is an extremely safe procedure and can be repeatedly applied to an eye. PMID- 19284324 TI - Fluocinolone inhibits VEGF expression via glucocorticoid receptor in human retinal pigment epithelial (ARPE-19) cells and TNF-alpha-induced angiogenesis in chick chorioallantoic membrane (CAM). AB - PURPOSE: The purpose of this study was to determine whether fluocinolone inhibits vascular endothelial growth factor (VEGF) expression in a retinal pigment epithelial cell line (ARPE-19) and TNF-alpha-induced angiogenesis in chick chorioallantoic membrane (CAM) assay. METHODS: The dose-dependent effect of fluocinolone (0.0001-1 microM) on VEGF secretion, VEGF mRNA expression, and cytotoxicity was determined in confluent monolayers of ARPE-19 cells using ELISA, RT-PCR, and MTT assay, respectively. The effect of a glucocorticoid receptor antagonist (RU486) on fluocinolone-mediated VEGF expression was determined. The effect of fluocinolone in inhibiting TNF-alpha-induced angiogenesis was determined using chick chorioallantoic membrane (CAM) assay. The dose-dependent effect of fluocinolone (0.0001-1 microM) in inhibiting 1% serum-stimulated ARPE 19 cell proliferation was determined using BrdU labeling assay. RESULTS: At concentrations devoid of cytotoxicity, fluocinolone inhibited VEGF secretion as well as mRNA expression in ARPE-19 cells. RU486 (1 microM) treatment prevented inhibition of VEGF secretion and VEGF mRNA expression by fluocinolone (0.1 microM). Fluocinolone (50 ng/egg) inhibited angiogenesis induced by TNF-alpha. The ARPE-19 cell proliferation was inhibited by fluocinolone in a dose-dependent manner. CONCLUSIONS: Fluocinolone inhibited VEGF expression in ARPE-19 cells via its glucocorticoid receptor activity. In addition, fluocinolone inhibited proliferation of ARPE-19 cells and TNF-alpha-induced angiogenesis in chorioallantoic membranes. PMID- 19284325 TI - Correlation of in vitro and in vivo kinetics of nitric oxide donors in ocular tissues. AB - In the eye, nitric oxide (NO) is involved in the regulation of intraocular pressure (IOP) and ocular blood flow. The main purpose of this study was to measure the kinetics of NO release from NO donors in ocular cells and tissues using in vivo and in vitro models and demonstrate the link between the kinetics of NO release with the functional effect, IOP. Nitric oxide release was measured in human ocular cells using a fluorescent dye, diaminofluorescein (DAF), following treatment with short-acting sodium nitroprusside (SNP) and longer acting S-nitroso-N-acetylpenicillamine (SNAP) NO donors. Both SNP and SNAP were also administered topically to rabbits; IOP was measured and levels of NO and cGMP were assessed as biomarkers over a time course in the aqueous humor (AH) and iris/ciliary body (ICB). Time- and concentration-dependent increases in NO level were produced by SNP and SNAP in human ocular cells. Both NO and cGMP levels appeared to be elevated following treatment with the aforementioned NO donors in rabbit ocular tissues. Transient IOP lowering was accompanied with these biochemical estimations in rabbits, with time of maximal effect being shifted to the right for longer-acting SNAP as compared with short-acting SNP. In vitro and in vivo NO/cGMP assay results displayed a correlation between short- and longer acting NO donors, discriminating their respective temporal actions in the eye. Due to their translatability, the in vitro DAF assay and in vivo NO fluorometric assay can therefore be potentially useful in screening novel NO donors with different temporal/kinetic profiles. PMID- 19284326 TI - Enlargement of the foveal avascular zone in diabetic retinopathy after adjunctive intravitreal bevacizumab (avastin) with pars plana vitrectomy. AB - PURPOSE: The aim of this study was to describe a case of angiographically documented foveal avascular zone (FAZ) enlargement after intravitreal bevacizumab following vitrectomy and intravitreal bevacizumab treatment of a diabetic patient. METHODS: This study was done as a case report. RESULTS: A 40-year-old diabetic man was treated with a pars plana vitrectomy and intravitreal bevacizumab for a nonclearing vitreous hemorrhage. Despite successful surgery, the postoperative best-corrected visual acuity was not improved. FAZ area had increased from 1.48 mm(2) preoperatively to 1.96 mm(2) postoperatively (24.5% increase). CONCLUSIONS: Although intravitreal bevacizumab is an effective modality for diabetic retinopathy, it may deteriorate pre-existing retinal ischemia. PMID- 19284327 TI - The use of voriconazole in the management of post-penetrating keratoplasty Paecilomyces keratitis. AB - PURPOSE: The aim of this study was to report successful treatment of Paecilomyces lilacinus keratitis with voriconazole. METHOD: Case series. RESULTS: Two patients with history of penetrating keratoplasty (PKP) presented with progressive P. lilacinus keratitis. Initial medical treatment failed to control the infection. The first patient received topical voriconazole at the early stage of infection. The second patient required therapeutic PKP and intracameral voriconazole was given at the end of the surgery. He developed presumed secondary endophthalmitis and was treated aggressively with intravitreal and systemic voriconazole. Both the patients responded well to the treatment and achieved complete recovery. CONCLUSIONS: These cases illustrated effectiveness and safety of various modes of voriconazole administration in treating P. lilacinus ocular infection. PMID- 19284328 TI - Comparative toxicity of preservatives on immortalized corneal and conjunctival epithelial cells. AB - PURPOSE: Nearly all eye drops contain preservatives to decrease contamination. Nonpreservatives such as disodium-ethylene diamine tetra-acetate (EDTA) and phosphate-buffered saline are also regularly added as buffering agents. These components can add to the toxicity of eye drops and cause ocular surface disease. To evaluate the potential toxicity of these common components and their comparative effects on the ocular surface, a tissue culture model utilizing immortalized corneal and conjunctival epithelial cells was utilized. METHODS: Immortalized human conjunctival and corneal epithelial cells were grown. At confluency, medium was replaced with 100 microL of varying concentrations of preservatives: benzalkonium chloride (BAK), methyl paraben (MP), sodium perborate (SP), chlorobutanol (Cbl), and stabilized thimerosal (Thi); varying concentrations of buffer: EDTA; media (viable control); and formalin (dead control). After 1 h, solutions were replaced with 150 microL of MTT (3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl tetrazonium bromide). After 4 h, solutions decanted, 100 microL of acid isopropanol added, and the optical density determined at 572 nm to evaluate cell viability. RESULTS: Conjunctival and corneal cell toxicity was seen with all preservatives. Depending upon concentration, BAK exhibited from 56% to 89% toxicity. In comparison, Cbl exhibited from 50% to 86%, MP from 30% to 76%, SP from 23% to 59%, and Thi from 70% to 95%. EDTA with minimal toxicity (from 6% to 59%) was indistinguishable from SP. CONCLUSIONS: Generally, the order of decreasing toxicity at the most commonly used concentrations: Thi (0.0025%) > BAK (0.025%) > Cbl (0.25%) > MP (0.01%) > SP (0.0025%) approximately EDTA (0.01%). Even at low concentration, these agents will cause some degree of ocular tissue damage. PMID- 19284329 TI - Intraocular pressure control among patients transitioned from latanoprost to travoprost at a Veterans Affairs Medical Center Eye Clinic. AB - PURPOSE: To evaluate intraocular pressure (IOP) control and the ocular adverse effects resulting from a large-scale transition from latanoprost to travoprost among patients at a Veterans Affairs Medical Center (VAMC) Eye Clinic. METHODS: Retrospective chart review of patients transitioned from latanoprost to travoprost after a revision of the drug formulary used by the VAMC in Houston, Texas. IOP control after changing medications and the incidence of ocular adverse effects attributed to travoprost were the main outcomes measured. For patients who were using IOP-lowering medications bilaterally, the worse eye was used for all IOP analyses. Long-term retention in IOP control plus a cost-saving analysis were presented as a secondary assessment. RESULTS: Five hundred ninety-nine (599) patients with 1,041 treated eyes were evaluated. Mean IOP was 15.86 +/- 4.15 mmHg among patients using latanoprost prior to the prostaglandin analog transition. After transitioning to travoprost, the mean IOP was 15.78 +/- 4.38 mmHg. The mean within-eye change in IOP in the worse eye when transitioned from latanoprost to travoprost was -0.07 +/- 3.27 mmHg (P = 0.5914). Twenty-four (24) patients (4%) experienced an ocular adverse effect while using travoprost. In the long-term retention analysis at 1 year, mean change in IOP from the time of the original change to travoprost was +0.21 +/- 3.71 mmHg (P = 0.2683). CONCLUSIONS: The large scale transition from latanoprost to travoprost maintained long-term IOP control. Only a small percentage of clinic patients experienced mild ocular adverse effects after being transitioned to the new prostaglandin analog. PMID- 19284330 TI - Sleep apnea--unmasking a nighttime health risk. PMID- 19284331 TI - Ear crop, tail dock policy not a radical departure, AVMA says. PMID- 19284332 TI - More on changing trends in practice management. PMID- 19284333 TI - The veterinarian's responsibility to recognize and report animal abuse. PMID- 19284334 TI - What is your diagnosis? Cardiogenic pulmonary edema. PMID- 19284335 TI - What is your neurologic diagnosis? Vestibular disease secondary to brain neoplasm. PMID- 19284336 TI - Anesthesia case of the month. Acute intraoperative tumor lysis. PMID- 19284337 TI - Theriogenology question of the month. Scoliosis. PMID- 19284338 TI - Income of US veterinarians, 2007. PMID- 19284339 TI - First rDNA construct approved for human use. PMID- 19284340 TI - Thoughts on canine dominance aggression. PMID- 19284342 TI - Evaluation of serum cardiac troponin I concentration in dogs with renal failure. AB - OBJECTIVE: To determine whether dogs with renal failure have higher serum cardiac troponin I (cTnI) concentrations than healthy dogs. DESIGN: Case-control study. ANIMALS: 31 dogs with renal failure and 51 healthy dogs. PROCEDURES: Serum concentrations of creatinine and cardiac troponin I, urine specific gravity, and systolic arterial blood pressure were measured for all dogs. Dogs underwent a standardized physical examination, and any dog with evidence of cardiovascular disease or other nonrenal disease was excluded from final analyses. Dogs were considered to be in renal failure when the serum creatinine concentration was >or= 3.0 mg/dL, urine specific gravity was between 1.007 and 1.030, and renal failure had been clinically diagnosed. RESULTS: Dogs with renal failure had significantly higher serum cTnI concentrations (median, 0.35 ng/mL) than did healthy dogs (0.20 ng/mL). The renal failure group also had a significantly higher median systolic blood pressure (156 mm Hg) than did healthy dogs (138 mm Hg), although serum cTnI concentration was not correlated with systolic blood pressure in dogs with renal failure. There was no significant difference in age between dogs with renal failure and healthy dogs, but dogs with renal failure had significantly higher serum creatinine concentration and lower urine specific gravity. CONCLUSIONS AND CLINICAL RELEVANCE: Although dogs with renal failure did not have overt clinical signs of cardiac disease, they had high serum cTnI concentrations, which may have been associated with subclinical cardiovascular disease. The cause of the high serum cTnI concentration in these dogs requires additional investigation. PMID- 19284343 TI - Signalment, history, and outcome of cats with gastrointestinal tract intussusception: 20 cases (1986-2000). AB - OBJECTIVE: To determine signalment, history, and outcome of cats with gastrointestinal tract intussusception and to identify physical examination, diagnostic imaging, surgical, histologic, and necropsy findings in affected cats. DESIGN: Retrospective case series. ANIMALS: 20 cats with intussusception. PROCEDURES: Medical records were evaluated for information on signalment; history; physical examination, diagnostic imaging, surgical, histologic, and necropsy findings; and outcome. RESULTS: Ten cats were < 1 year old, and 9 were >or= 6 years old. Anorexia (14/17), lethargy (12/17), and vomiting (12/17) were the most common reasons for examination. Dehydration (13/18), poor body condition (12/18), signs of abdominal pain (8/18), and an abdominal mass (8/18) were the most common physical examination findings. Abdominal radiography revealed intestinal obstruction in all 10 cats in which it was performed; abdominal ultrasonography revealed intussusception in all 7 cats in which it was performed. The most common intussusception was jejuno-jejunal (8/20), and no intussusceptions were found proximal to the duodenum. Eleven of 13 cats that underwent laparotomy required intestinal resection and anastomosis. Histologic examination revealed intestinal lymphoma or inflammatory bowel disease in 7 of 8 cats >/= 6 years old and idiopathic intussusception in 7 of 8 cats < 1 year old. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that in cats, intussusception has a bimodal age distribution, is most commonly jejuno-jejunal, often requires surgical resection and anastomosis, is often associated with alimentary lymphoma or inflammatory bowel disease in older cats, and is readily diagnosed by means of ultrasonography. PMID- 19284345 TI - Comparison of clinical, microbiologic, and clinicopathologic findings in horses positive and negative for Clostridium difficile infection. AB - OBJECTIVE: To compare clinical, microbiologic, and clinicopathologic findings among horses infected with Clostridium difficile that had toxin A in their feces, horses with evidence of C difficile infection that were negative for toxin A in their feces, and horses with diarrhea that were negative for C difficile infection. DESIGN: Cross-sectional study. ANIMALS: 292 horses and foals with diarrhea. PROCEDURES: Feces were submitted for microbial culture and tested for the C difficile antigen glutamate dehydrogenase and for toxin A with a commercial ELISA. RESULTS: Horses with toxin A in their feces had higher band neutrophil count, rectal temperature, hospitalization time prior to the onset of diarrhea, and total hospitalization time than did horses without evidence of C difficile infection, and 32 of the 33 (97%) horses with toxin A in their feces had received antimicrobials prior to the onset of diarrhea. Horses with toxin A in their feces had a significantly higher mortality rate than did horses negative for toxin A in their feces. Sensitivity and specificity of the ELISA for detection of C difficile antigen were 93% and 88%, when assay results were compared with results of microbial culture following direct plating, and 66% and 93%, when assay results were compared with results of microbial culture following broth enrichment. CONCLUSIONS AND CLINICAL RELEVANCE: Results provided some evidence that horses positive for toxin A had more severe clinical disease than did horses with evidence of C difficile infection that were negative for toxin A and horses with diarrhea without evidence of C difficile infection. PMID- 19284346 TI - Evaluation of factors affecting serum IgG concentrations in bottle-fed calves. AB - OBJECTIVE: To determine the effect of time interval from birth to first colostrum feeding on colostrum intake and serum IgG concentration and the effect of varying colostral volume intake and colostral IgG concentration on the probability of failure of passive transfer (FPT) in bottle-fed calves. DESIGN: Randomized controlled study. ANIMALS: 104 calves. PROCEDURES: Equal numbers of calves were randomly assigned to groups and fed 3 L of their dam's colostrum at 1, 2, 3, or 4 hours after birth by use of a nipple bottle. Calves were allowed to feed for 15 minutes, and intake was recorded. A second 3-L bottle feeding of colostrum was offered at 12 hours of age. RESULTS: 17.2% of calves ingested 3 L of colostrum at the first feeding and 3 L at 12 hours of age. Calf age, up to 4 hours, had no significant effect on the calf's ability to ingest colostrum or on 48-hour serum IgG concentration. Colostral intake at 1, 2, 3, or 4 hours had no effect on intake at the second feeding. Probability of FPT in calves ingesting 3 L at both feedings was < 0.05. CONCLUSIONS AND CLINICAL RELEVANCE: Allowing calves fed by nipple bottle to ingest as much colostrum as they can within 4 hours after birth and at 12 hours of age substantially reduced the probability of FPT. Bottle-fed calves that do not ingest 3 L of colostrum within the first 4 hours after birth should be targeted for oroesophageal intubation. PMID- 19284347 TI - Evaluation of prepartum serum cholesterol and fatty acids concentrations as predictors of postpartum retention of the placenta in dairy cows. AB - OBJECTIVE: To identify serum biochemical and hematologic variables, as measured in the week before parturition, that predict postpartum retention of the placenta (RP) in dairy cows. DESIGN: Retrospective cohort study. ANIMALS: 1,038 cows in 20 commercial dairy herds. PROCEDURES: Serum concentrations of fatty acids (FAs), beta-hydroxybutyrate, cholesterol, glucose, urea, and calcium and blood leukocyte, neutrophil, lymphocyte, monocyte, and eosinophil counts were determined. These variables were evaluated for an association with development of RP by use of a multivariate logistic regression model. Parity, season of parturition, existence of twins or dystocia, body condition score, and vitamin E treatment were included in the model as covariates. RESULTS: High serum concentrations of cholesterol and FAs were associated with an increased odds of RP. There was a 5% relative increase in the odds of RP for each 0.1 mmol/L increase in cholesterol or FAs concentration in the week before parturition. Season of parturition and twinning were also identified as risk factors. CONCLUSIONS AND CLINICAL RELEVANCE: These associations indicated that prepartum energy metabolism contributes to the development of RP. Serum concentrations of cholesterol and FAs may be useful to identify cows with a metabolic abnormality or energy imbalance that might predispose them to RP and should be interpreted in conjunction with clinical risk factors such as twinning, dystocia, or parturient paresis. PMID- 19284348 TI - Degenerative disease of the distal interphalangeal joint and sesamoid bone in calves: 9 cases (1995-2004). AB - OBJECTIVE: To evaluate the clinical signs, diagnosis, treatment, and outcome associated with distal interphalangeal joint (DIJ) degenerative disease in calves. DESIGN: Retrospective case series. ANIMALS: 9 calves with degenerative disease of the DIJ. PROCEDURES: Medical records of calves with lameness for which the cause had been localized to the distal aspect of a limb but without evidence of trauma or sepsis were included. Signalment and results of physical, lameness, radiographic, and postmortem examinations were recorded. Outcomes of medical and surgical treatments were evaluated. RESULTS: All 9 calves had forelimb lameness (4 bilaterally). The medial digit was affected in 12 limbs and the lateral digit was affected in 9 limbs. Radiographic examination revealed bridging osteoproliferation along the abaxial and palmar aspects of affected DIJs (18/21), subchondral cyst-like lucencies (19/21), and sclerosis (16/21). The DIJ was resected in 3 calves (7 joints), resulting in resolution of lameness within 3 months. Medical management was attempted in 4 calves (6 joints), and 2 calves (6 joints) did not receive treatment; 5 calves had a normal stance within 5 months. Results of postmortem computed tomographic and histologic examination were in accordance with degenerative joint disease processes. CONCLUSIONS AND CLINICAL RELEVANCE: Degenerative disease of the DIJ should be considered in calves with insidious, progressive forelimb lameness. Although the etiology of this process remains undetermined, results of radiographic, computed tomographic, and histologic examinations suggested an underlying developmental anomaly, such as osteochondrosis. Surgical treatment appeared promising on the basis of postoperative resolution of lameness in all surgically treated calves. PMID- 19284350 TI - An endoscopic method for identifying sex of hatchling Chinese box turtles and comparison of general versus local anesthesia for coelioscopy. AB - OBJECTIVE: To establish a safe and effective endoscopic method for visualizing the gonads and identifying the sex of hatchling Chinese box turtles and to compare the effects of general versus local anesthesia during coelioscopy. DESIGN: Clinical trial. ANIMALS: 58 hatchling Chinese box turtles (Cuora flavomarginata). PROCEDURES: Turtles were randomly assigned to be anesthetized with a mixture of ketamine, medetomidine, and morphine (n = 29) or to receive local anesthesia with lidocaine in the prefemoral region (29). Coelioscopy was performed with a rigid 1.9- or 2.7-mm telescope following insufflation with sterile lactated Ringer's solution. Ease of endoscopic sex identification and quality of anesthesia were scored. Body weights were recorded before and 7 and 14 days after surgery. RESULTS: Gonads were easily visualized and sex was easily identified in all 58 turtles without complications. Endoscopy scores and pre- and postoperative weights did not differ significantly between groups. However, anesthesia scores were significantly worse for animals that received local anesthesia alone, compared with those that underwent general anesthesia. All anesthetized turtles recovered within 21 minutes after administration of the reversal agents, atipamezole and naloxone. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that coelioscopy with a rigid endoscope and lactated Ringer's solution for insufflation was a safe and effective method for identifying the sex of hatchling Chinese box turtles. General anesthesia was effective and effects were rapidly reversible; local anesthesia with lidocaine alone was considered insufficient for coelioscopy. PMID- 19284351 TI - MicroRNA profiling of human-induced pluripotent stem cells. AB - MicroRNAs (miRNAs) are a newly discovered endogenous class of small noncoding RNAs that play important posttranscriptional regulatory roles by targeting mRNAs for cleavage or translational repression. Accumulating evidence now supports the importance of miRNAs for human embryonic stem cell (hESC) self-renewal, pluripotency, and differentiation. However, with respect to induced pluripotent stem cells (iPSC), in which embryonic-like cells are reprogrammed from adult cells using defined factors, the role of miRNAs during reprogramming has not been well-characterized. Determining the miRNAs that are associated with reprogramming should yield significant insight into the specific miRNA expression patterns that are required for pluripotency. To address this lack of knowledge, we use miRNA microarrays to compare the "microRNA-omes" of human iPSCs, hESCs, and fetal fibroblasts. We confirm the presence of a signature group of miRNAs that is up regulated in both iPSCs and hESCs, such as the miR-302 and 17-92 clusters. We also highlight differences between the two pluripotent cell types, as in expression of the miR-371/372/373 cluster. In addition to histone modifications, promoter methylation, transcription factors, and other regulatory control elements, we believe these miRNA signatures of pluripotent cells likely represent another layer of regulatory control over cell fate decisions, and should prove important for the cellular reprogramming field. PMID- 19284353 TI - Risk of anemia and transfusion triggers: implications for bloodless care. AB - BACKGROUND: Blood transfusion is a widely used treatment that is expensive and carries a small degree of risk. The level of anemia that patients tolerate and the indications for blood transfusion are controversial. METHODS: Review of pertinent English language literature. RESULTS: The risk of blood transfusion from viral transmission is very low. The risk of anemia increases as hemoglobin concentration decreases. The risk is very high when the hemoglobin concentration is below 6 g/dL. The risk of death or serious morbidity increases more rapidly in patients with cardiovascular disease than patients without cardiovascular disease. There is only one adequately powered clinical trial that demonstrates that a 7 g/dL transfusion threshold is as safe, and in some subgroups, safer than a 10 g/dL threshold. CONCLUSIONS: A restrictive transfusion threshold (7 g/dL) is recommended in most patients. In patients with cardiovascular disease, a 9-10 g/dL may be advisable, but definitive guidelines await further clinical trials. However, as with all such treatments, the decision to transfuse should be based on individual assessment of patients' signs, symptoms, and co-morbidity. PMID- 19284354 TI - The current infectious risks of transfusions. AB - BACKGROUND: Red blood cell transfusions have been widely associated with the transmission of human immunodeficiency virus (HIV) especially prior to 1985 when testing for HIV was not available. Currently the blood supply is much safer because of very sensitive testing for HIV as well as hepatitis C. However, new infectious agents emerge constantly and pose a threat to our blood supply. Current and potential threats are reviewed in this manuscript. METHODS: Review of pertinent English language literature. RESULTS: Transmission of infectious agents from blood transfusion is rare. Protozoans pose one the greatest threats because there are no assays available that can reliably detect their presence and because of worldwide travel. Viruses potentially pose a significant threat to the blood supply. The West Nile virus has recently been transmitted by a blood transfusion that was negative by assay for WNV. Severe acute respiratory syndrome (SARS) poses a threat to the blood supply but has never been transmitted in a transfusion. Prions also have been transmitted by a blood transfusion. CONCLUSIONS: Several infectious agents pose potential threats. The current risk of a blood transfusion is very low but the potential threat is ever present. PMID- 19284355 TI - Rationale for blood conservation. AB - BACKGROUND: Exposure of patients to allogeneic blood transfusion can be minimized or avoided by the systematic use of multiple blood conservation techniques. Current use of these technologies is variable. METHODS: Review of pertinent English language literature. RESULTS: Enthusiasm for preoperative autologous blood donation (PAD) has declined considerably, perhaps due to increased cost and inconvenience to patients. Acute normovolemic hemodilution (ANH) has several practical advantages over PAD, but has not become generally accepted as a blood conservation strategy. Erythropoietin, iron, and artificial oxygen carriers are pharmacologic alternatives. CONCLUSIONS: Pharmacologic stimulation of erythropoiesis offers substantial potential to progress toward a goal of bloodless medicine. The potential of artificial blood substitutes is still being defined. PMID- 19284356 TI - Transfusion in the intensive care unit. AB - BACKGROUND: Anemia is a common complication of critical illness. Because tissue hypoxia is a prominent factor in the development of organ dysfunction in the critically ill, conventional wisdom has argued that the transfusion of packed red blood cells can attenuate tissue hypoxia and so improve outcome. METHODS: Review of pertinent English-language literature. RESULTS: The empiric evidence supporting the benefit of transfusion to treat tissue hypoxia is sparse; indeed, a body of recent work suggests that moderate anemia is not only well-tolerated by the critically ill patient, it is associated with improved clinical outcomes. The primary biologic rationale for transfusion of the critically ill is to maximize oxygen delivery to tissues. However, because of reflex compensatory mechanisms, and because of alterations in microvascular flow and endothelial permeability, the impact of transfusion is much less than would be predicted. Retrospective studies suggest that transfusion is immunosuppressive, and associated with an enhanced infectious risk. The large Transfusion Requirements in Critical Care (TRICC) Trial conducted by the Canadian Critical Care Trials Group demonstrated reduced mortality and organ dysfunction when a transfusion trigger of 7 g/dL is used, without an increase in infectious complications. CONCLUSIONS: A conservative transfusion strategy appears safe in nearly all critically ill patients without active hemorrhage, including patients with cardiovascular disease. Whether a lower transfusion threshold could be adopted is unknown. PMID- 19284357 TI - Iron and the risk of infection. AB - BACKGROUND: During bacterial infection, pathogen and host compete for iron (Fe). The inflammatory response associated with infection shifts Fe from the circulation into storage, resulting in hypoferremia and iron-deficient erythropoiesis, and ultimately contributing to the anemia of inflammation. METHODS: In this article, we review the mechanisms of Fe acquisition and sequestration. Bacteria employ both membrane-bound transferrin receptors and high affinity iron-binding proteins called siderophores to acquire Fe. Humans utilize the iron-binding proteins lactoferrin, transferrin, and ferritin to move Fe away from sites of infection and into storage. Synthesis and action of these proteins are regulated by inflammatory cytokines. RESULTS: Iron overload leads to inhibition of IFN-gamma, TNF-alpha, IL-12, and nitric oxide formation as well as impairment of macrophage, neutrophil, and T-cell function. Injection of Fe into mice and rats markedly increases the virulence of several pathogens. Studies in hemodialysis patients have documented an association between infection and increased ferritin concentration as a surrogate marker for Fe overload. CONCLUSIONS: Humans respond to infection with inflammatory cytokine-induced hypoferremia. This association, as well as the growing literature linking Fe to both impaired immunity and heightened microbial virulence, calls into question the value of Fe supplementation during inflammation and infection. PMID- 19284358 TI - Acute normovolemic hemodilution. AB - BACKGROUND: The inherent risks of blood transfusion have led to concerted efforts to find alternatives to allogeneic blood transfusion. Among these alternatives are supplementation with iron and treatment with erythropoietin, autologous pre donation before major elective surgery, intraoperative blood salvage (particularly for emergency cases or when major blood loss is unanticipated), acute normovolemic hemodilution, and artificial blood substitutes. METHODS: Review of pertinent English-language literature. RESULTS: Autologous pre-donation of the patient's own blood, even if supplemented with iron and erythropoietin, is increasingly out of favor for several reasons. Autologous pre-donation is an added out-of-pocket expense for the patient. The major cause of incompatible blood transfusion, namely clerical error, is not eliminated. Moreover, autologous pre-donated blood is not released to the general blood supply; therefore, autologous blood that is not returned to the patient by infusion is wasted. Acute normovolemic hemodilution is a safe alternative that eliminates the risk of mistransfusion, minimizes the shed red cell mass when bleeding occurs, and reduces the risk of perioperative transfusion as shown in patients undergoing radical prostatectomy. CONCLUSIONS: Acute normovolemic hemodilution achieves equivalent outcomes at lower cost compared with autologous pre-donation. Patient safety is increased, and if pharmacologic strategies are combined with acute normovolemic hemodilution, allogeneic blood transfusion may be eliminated entirely. PMID- 19284359 TI - Treatment options for restless legs syndrome. AB - BACKGROUND: Restless legs syndrome (RLS) is a common condition characterized by dysesthesia and an urge to move. Predominantly, symptoms occur at rest in the evening or at night, and they are alleviated by moving the affected extremity or by walking. Although the etiopathogenesis of RLS is still unknown, the rapid and dramatic improvement of RLS with dopaminergic agents suggests that dopaminergic system dysfunction may be a basic mechanism. Dopaminergic agents are the best studied agents, and are considered first-line treatment of RLS. OBJECTIVE: To review all options for the treatment of RLS, including the non-pharmacological ones. METHODS: The treatment suggestions are based on evidence from studies published in peer-reviewed journals, or upon a comprehensive review of the medical literature. RESULTS/CONCLUSION: Extensive data are available for levodopa and dopamine agonists, especially for pramipexole and ropinirole. Pharmacological treatment should be limited to those patients who suffer from clinically relevant RLS with impaired sleep quality or quality of life. A treatment on demand is a clinical need in RLS cases that present intermittent symptoms. PMID- 19284361 TI - Treatment strategies for a patient with rheumatoid arthritis and hepatitis C. AB - BACKGROUND: The poor prognosis of rheumatoid arthritis (RA) can be aggravated by the concomitant presence of chronic hepatitis C virus (HCV) infection and there are no guidelines for the treatment of patients affected by both conditions. OBJECTIVE: To propose new therapeutic strategies for patient affected by RA and concomitant HCV chronic infection. METHODS: Review of the literature on the usage of cyclosporine-A (CsA) and anti-tumour-necrosis-factor (TNF)-alpha agents for the treatment of patients affected by RA and HCV. RESULTS/CONCLUSION: CsA exerts an inhibitory effect on HCV replication and it is safe in patients affected by RA and HCV. Anti-TNF-alpha agents are safe and efficacious in patient with RA and HCV. Anti-TNF-alpha and CsA can be safely given in combination in RA patients with HCV infection. PMID- 19284360 TI - The use of cephalosporins for gonorrhea: the impending problem of resistance. AB - Gonorrhea remains an important clinical and public health problem throughout the world. Gonococcal infections have historically been diagnosed by Gram stain and culture but are increasingly diagnosed through nucleic acid tests, thereby eliminating the opportunity for antimicrobial susceptibility testing. Gonococcal infections are typically treated with single-dose therapy with an agent found to cure > 95% of cases. Unfortunately, the gonococcus has repeatedly developed resistance to antimicrobials including sulfonamides, penicillin, tetracyclines and fluoroquinolones. This has now left third-generation cephalosporins as the lone class of antimicrobials recommended as first-line therapy for gonorrhea in some regions. However, resistance to oral third-generation cephalosporins has emerged and spread in Asia, Australia and elsewhere. The mechanism of this resistance seems to be associated with a mosaic penicillin binding protein (penA) in addition to other chromosomal mutations previously found to confer resistance to beta-lactam antimicrobials (ponA, mtrR, penB, pilQ). Few good options exist or are in development for treating cephalosporin-resistant isolates, as most have had multidrug resistance. Preventing the spread of resistant isolates will depend on ambitious antimicrobial management programs, strengthening and expanding surveillance networks, and through effective sexually transmitted disease control and prevention. PMID- 19284362 TI - Multitarget drugs: the present and the future of cancer therapy. AB - Target therapies for the treatment of human cancers have revolutionized the concept of oncological medicine. This type of therapeutic approach is directed to the inhibition of molecular targets that play a pivotal role in tumor progression -- such as tyrosine kinase receptors (TKIs) controlling cell proliferation and survival -- mainly by means of compounds able to block their activity. In the beginning, the aim of target therapies was specifically to hit a single molecule expressed in neoplastic cells. Now the prevailing idea is that inhibiting both cancer cells and cells of the stroma supporting the tumor would gain better results in fighting the disease. Therefore, the single-target therapy is fading in favor of a multitarget approach and the new generation of TKIs is selected on the basis of their ability simultaneously to target different molecules. This review summarizes the molecular basis of multitarget therapies and the most relevant results obtained in different cancer types. PMID- 19284363 TI - Thromboprophylaxis and obstetric management of the antiphospholipid syndrome. AB - There is consensus in treating patients with antiphospholipid syndrome (APS) and first venous thrombosis with oral anticoagulation to a target international normalized ratio of 2.0 - 3.0. A recent systematic review recommended a target international normalized ratio > 3.0 in the group of patients with APS and arterial thrombosis. The approach for women with obstetric manifestations of APS is based on the use of aspirin plus heparin. In addition, we summarize the evidence-based information about management of catastrophic APS and some difficult cases, such as 'seronegative' APS, patients who do not display formal classification criteria for APS, and recurrent thrombotic events despite optimal anticoagulation. PMID- 19284364 TI - An update on the treatment options for focal segmental glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) is not a disease but a lesion initially affecting the podocyte. Various factors may induce 'secondary' FSGS, including defects in molecules that contribute to the podocyte slit diaphragm permselectivity to albumin. They do not represent indications for immunosuppression and require symptomatic treatment only, comprising angiotensin 2 and endothelin antagonists. Primary (idiopathic) FSGS is possibly but not certainly of immunologic origin, owing to an elusive glomerular permeability factor (GPF), explaining relapse on a renal transplant and justifying an immunosuppressive treatment. The best prognostic feature of primary nephrotic FSGS is its response to corticosteroids. Alkylating agents are mostly ineffective in steroid-resistant forms. An association of corticosteroids and cyclosporine A (CsA) remains the mainstay of treatment, with a good tolerability when CsA dosage is low. A definite advantage of tacrolimus on CsA has not yet been established. Sirolimus appears ineffective and potentially harmful. Azathioprine is not indicated. A number of mostly uncontrolled trials indicate that mycophenolate mofetil might find an adjunctive place in the treatment. Plasmapheresis is of no avail outside the special case of relapse in a transplanted kidney. Immunoabsorption of the GPF has not led to practical treatment options. Anecdotal reports on rituximab are as yet too few to determine whether this monoclonal anti CD20 antibody will find a place in the treatment of primary FSGS. PMID- 19284365 TI - Antiemetic control: toward a new standard of care for emetogenic chemotherapy. AB - Chemotherapy-induced nausea and vomiting (CINV) is associated with a significant deterioration in quality of life. The emetogenicity of the chemotherapeutic agents, repeated chemotherapy cycles, and patient risk factors significantly influence CINV. 5-hydroxytryptamine-3 (5-HT(3)) receptor antagonists plus dexamethasone have significantly improved the control of acute CINV, but delayed CINV remains a significant clinical problem. Two new agents, palonosetron and aprepitant, have been approved for the prevention of both acute and delayed CINV. Palonosetron is a second-generation 5-HT(3) receptor antagonist with a longer half-life and a higher binding affinity than first-generation 5-HT(3) receptor antagonists. Aprepitant is the first agent available in the new drug class of neurokinin-1 (NK-1) receptor antagonists. Casopitant is another NK-1 receptor antagonist that is under review by the FDA after recent completion of Phase III clinical trials. The introduction of these new agents has generated revised antiemetic guidelines for the prevention of CINV. Future studies may consider the use of palonosetron, aprepitant and casopitant with other antiemetic agents (olanzapine, gabapentin, cannabinoids) in moderately and highly emetogenic chemotherapy, as well as in the clinical settings of multiple-day chemotherapy and bone marrow transplantation. PMID- 19284366 TI - Pharmacotherapy of recurrent respiratory papillomatosis: an expert opinion. AB - BACKGROUND: Recurrent respiratory papillomatosis is caused by the human papillomavirus types (HPV) 6 and 11. It affects both children and adults. In a small number of cases, the disease can be very aggressive causing significant morbidity and possibly death. Surgical therapy is the primary treatment but in patients with aggressive disease, adjunctive therapy is initiated. The majority of these adjuncts center on immunomodulation, disruption of molecular signaling cascades or interruption of viral replication to help decrease the severity of the disease. Recently, a preventative vaccine has become available but data on its effectiveness will be at least a decade away. In the mean time, researchers are examining other vaccination strategies in the fight against HPV disease. OBJECTIVE: We will review the following pharmacotherapies used in the adjunct treatment of RRP: interferon, acyclovir, ribivirin, cidofovir, COX-2 inhibitors, retinoids, anti-reflux medications, zinc, indole-3-carbinol, therapeutic/preventative vaccines. METHODS: This is a review paper. Utilizing Medline and Pubmed from 1966 to present, the key words as well as the above listed adjunct treatments were searched for relevant papers. CONCLUSION: The evidence supporting each of these adjuncts varies with a majority having only case reports or cases-series to support their use. However, there is hope on the horizon with regard to the HPV vaccine and its potential to prevent future transmission of this disease. PMID- 19284367 TI - Insulin Glargine: a review 8 years after its introduction. AB - Insulin Glargine was the first long-acting insulin analog produced by recombinant DNA technology, approved for use by the US FDA in April 2000 and by the European Agency for the Evaluation of Medicinal Products in June, 2000. It has become the most widely used insulin in the USA owing to its long duration of action without a pronounced peak. The principal advantage of insulin Glargine over neutral protamine Hagedorn (NPH) insulin is in a lower frequency of hypoglycemic reactions, thus affording improved safety. It is used in both type 1 and type 2 diabetes, usually as a single daily dose. In type 2 patients, it is often the first insulin introduced as a single daily dose. Although insulin Glargine is typically administered as a single nighttime dose, it can be given in the morning or at any other time convenient for the patient. In labile type 1 diabetes, it is often most effective given as two daily injections. In obese, insulin-resistant patients, it may be best to administer insulin Glargine in two separate doses, owing to the high volumes of injected insulin required. Insulin Glargine does not treat postprandial hyperglycemia. It is necessary to supplement with short-acting insulin at mealtimes to control glucose surges after meals. Insulin Glargine is effective in hospitalized and postsurgical patients on account of its lack of pronounced insulin peaks and long duration of action. Although there is considerable use of Glargine in pregnant diabetic women, there is no definitive study to confirm its benefits. Insulin Glargine is thought to coprecipitate supplementary short-acting insulins when co-administered in the same syringe. Therefore, more injections are typically needed in the usual treatment regimen for insulin requiring diabetes. In many cases, constant basal insulin levels may be achieved with multiple overlapping doses of NPH insulin given together with short-acting insulin at mealtimes. Such a therapy may be less costly, but the major advantage of insulin Glargine remains the greater safety of a lower frequency of hypoglycemic reactions. PMID- 19284368 TI - UPLIFT Study: the effects of long-term therapy with inhaled tiotropium in chronic obstructive pulmonary disease. Evaluation of: Tashkin DP, Celli B, Senn S et al.: a 4-year trial of tiotropium in chronic obstructive pulmonary disease. N Engl J Med (2008) 359(15):1543-1554. AB - BACKGROUND: In chronic obstructive pulmonary disease (COPD), inhaled tiotropium bromide, a long-acting anticholinergic, has been shown to exert a sustained bronchodilator effect and to be superior to ipratropium bromide, a short-acting formulation of the same pharmacological class. OBJECTIVE: To discuss the effects of long-term therapy with tiotropium in COPD. METHODS/RESULTS: Analysis of efficacy and safety data on tiotropium from a 4-year randomized placebo controlled study performed in moderate to very severe COPD patients. Tiotropium was found to reduce significantly COPD-related morbidity, to improve health related quality of life (HRQoL) irrespective of disease severity and to slow significantly lung function decline in patients not using inhaled corticosteroids or other long-acting bronchodilators. The safety profile -- and in particular cardiovascular safety -- of tiotropium was good. CONCLUSIONS: Tiotropium bromide, alone or in combination with other inhaled therapies, can maintain an adequate control of COPD on a long-term basis. PMID- 19284369 TI - State-of-the-art imaging for detecting cancer in the clinic. PMID- 19284370 TI - Development of novel drugs targeting inhibitors of apoptosis. PMID- 19284372 TI - Infection and cancer: biology, therapeutics and prevention. PMID- 19284373 TI - YB-1 prevents apoptosis via the mTOR/STAT3 pathway in HER-2-overexpressing breast cancer cells. AB - Evaluation of: Lee C, Dhillon J, Wang MY et al.: Targeting YB-1 in HER-2 overexpressing breast cancer cells induces apoptosis via the mTOR/STAT3 pathway and suppresses tumor growth in mice. Cancer Res. 68 (21), 8661-8666 (2008). The transcription factor Y-box binding protein (YB)-1 is highly expressed in breast cancer cells and is strongly linked with breast cancer patient prognosis. In this paper, siRNA knockdown of YB-1 was used to investigate breast cancer cell proliferation. Six breast cancer cell lines that either overexpress HER-2 or were triple negative demonstrated growth inhibition following YB-1 knockdown. In particular, YB-1 knockdown induced apoptosis in BT-474-m1 and Au565 cells. Knockdown of YB-1 also decreased phosphorylation of STAT3S727, ERK1/2T202/Y204, mTORS2448 and total mTOR expression. When STAT3 was knocked down by siSTAT3, apoptosis was induced and constitutively active phosphorylated STAT3 was found to rescue YB-1-induced apoptosis. Furthermore, YB-1 knockdown remarkably suppressed colony formation in a soft agar assay, while delayed tumor formation was observed in mice. YB-1 knockdown inhibited cell growth and it is thought to involve induction of apoptosis via the mTOR/STAT3 intracellular signaling pathway. YB-1 is a promising molecular target for HER-2-overexpressing or triple-negative breast cancer cells. PMID- 19284374 TI - Localized squamous cell carcinoma of the esophagus: bimodality or trimodality approach? AB - Evaluation of: Lorenzen S, Brucher B, Zimmermann F et al.: Neoadjuvant continuous infusion of weekly 5-fluorouracil and excalating doses of oxaliplatin plus concurrent radiation in locally advanced oesophageal squamous cell carcinoma: results of a Phase I/II trial. Br. J. Cancer 99, 1020-1026 (2008). In this article, we summarize Lorenzen's study and briefly describe some of the landmark studies in preoperative chemoradiation in both squamous cell carcinoma and adenocarcinoma of the esophagus. Incorporation of oxaliplatin into the neoadjuvant regimen has been reported by several groups. The results of histological response varied from study to study. Lorenzen and colleagues reported a reasonably high histological response rate with very manageable toxicities in squamous cell carcinoma of the esophagus. However, as we know, squamous cell carcinoma is generally more responsive to chemoradiation. Future studies may be warranted to assess the efficacy of this regimen in a larger cohort. Nonetheless, in the era of targeted therapy, addition of targeted agents to future preoperative chemoradiation regimens should be considered. At present time, the result of RTOG Phase III study integrating cetuximab in the preoperative regimen is awaited. PMID- 19284375 TI - Telomerase structure paves the way for new cancer therapies. AB - Inappropriate activation of a single enzyme, telomerase, is associated with the uncontrollable proliferation of cells observed in as many as 90% of all of human cancers. Since the mid-1990s, when telomerase activity was detected in human tumors, scientists have eyed the enzyme as an ideal target for developing broadly effective anticancer drugs. One of the missing links in the effort to identify such therapies has been the high-resolution structure of the enzyme, a powerful tool used for the identification and development of clinical drugs. A recent structure of the catalytic subunit of teleomerase from the Skordalakes laboratory, a major advancement in the field of telomeres, has opened the door to the development of new, broadly effective cancer drugs, as well as anti-aging therapies. Here we present a brief description of telomerase biology, current efforts to identify telomerase function modulators and the potential importance of the telomerase structure in future drug development. PMID- 19284376 TI - Therapeutic potential of RNA interference in drug-resistant cancers. AB - Resistance including multidrug resistance to chemotherapy is a common clinical problem in patients suffering from cancer. Multidrug resistance is often mediated by overexpression of transmembrane xenobiotic transport molecules belonging to the superfamily of ATP-binding cassette (ABC)-transporters. Inhibition of ABC transporters by low-molecular weight compounds in cancer patients has been extensively investigated in clinical trials, but the results have been disappointing. Thus, alternative experimental therapeutic strategies for overcoming multidrug resistance are under investigation. These include the application of RNA interference (RNAi) technology. Various RNAi strategies were applied to reverse multidrug resistance in different tumor models in vitro and in vivo. Results and conclusions of these RNAi studies as well as their potential impact for the development of potential RNAi therapeutics will be discussed. PMID- 19284378 TI - Prognostic markers and targeted therapies for renal cell carcinoma. AB - Management of advanced renal cell carcinoma remains a persistent clinical challenge with high morbidity and mortality for a large proportion of patients. Until recently, available medical immunotherapy regimens yielded a therapeutic response in only 20% of patients. Advances in the understanding of molecular mechanisms of renal cell carcinoma have led to a rapidly expanding body of work exploring biomarkers for the disease and targeted therapeutics. We review current investigations into biomarkers and novel therapies for renal cell carcinoma, discuss the concept of anticancer vaccines, and propose a novel target for anticancer vaccine development. PMID- 19284377 TI - Promising novel immunotherapies and combinations for prostate cancer. AB - The field of therapeutic cancer vaccines is currently in a state of active preclinical and clinical investigation, and certain novel therapies involving tumor immunotherapy have recently come to the forefront of prostate cancer research. While no therapeutic cancer vaccine has yet been approved by the US FDA, recent findings have demonstrated that new paradigms of combination therapies involving vaccines, employed in clinical trials with appropriate design and end points, may ultimately lead to cancer vaccines being used to treat various malignancies. Several characteristics of prostate cancer make it an ideal target for immunotherapy. Its relative indolence allows sufficient time to generate immune responses, which usually take weeks or months to mount. In addition, prostate cancer-associated antigens direct the immune response to prostate cancer cells, thus sparing normal tissue. This review focuses on the future of promising new vaccines and novel perspectives in the treatment of prostate cancer. PMID- 19284379 TI - Intracranial ependymoma: factors affecting outcome. AB - Ependymomas account for 2-9% of all neuroepithelial tumors, amounting to 6-12% of all intracranial tumors in children and up to 30% of those in children younger than 3 years. Recent findings provide evidence that intracranial and spinal ependymomas share similar molecular profiles with the radial glia of their corresponding locations. The management of intracranial ependymoma is still not optimal. The 5-year progression-free survival for children with ependymoma ranges between 30 and 50% with a worse prognosis for patients with residual disease after surgery. The prognostic relevance of most factors are still being debated. Recent studies, in which the current WHO classification criteria were applied, reported the relationship between histological grade and outcome. Biomolecular studies have identified that gain of 1q25 and EGFR overexpression correlate to poor prognosis, whereas low expression of nucleolin correlated with a favorable outcome. Ependymomas have been considered a 'surgical disease', where completeness of excision can be reached in approximately half of the cases. At present the standard treatment is radiation therapy for all patients after gross total or near-total resection. For high-risk patients, with residual tumor, an interesting, although experimental, approach could be chemotherapy followed by secondary surgery and postoperative conformal irradiation. PMID- 19284380 TI - Pathogenesis of choriocarcinoma: clinical, genetic and stem cell perspectives. AB - Choriocarcinoma is a unique malignant neoplasm composed of mononuclear cytotrophoblasts and multinucleated syncytiotrophoblasts that produce human chorionic gonadotrophin. Choriocarcinoma can occur after a pregnancy, as a component of germ cell tumors, or in association with a poorly differentiated somatic carcinoma, each with distinct clinical features. Cytogenetic and molecular studies, predominantly on gestational choriocarcinoma, revealed the impact of oncogenes, tumor suppressor genes and imprinting genes on its pathogenesis. The role of stem cells in various types of choriocarcinoma has been studied recently. This review will discuss how such knowledge can enhance our understanding of the pathogenesis of choriocarcinoma, enable exploration of novel anti-choriocarcinoma targeted therapy and possibly improve our insight on embryological and placental development. PMID- 19284381 TI - Pleomorphic lobular carcinoma of the breast: molecular pathology and clinical impact. AB - Pleomorphic lobular carcinoma of the breast is a recently described morphological variant of classic invasive lobular carcinoma with an aggressive behavior. Morphologically, pleomorphic lobular carcinoma elicits a similar pattern of infiltrative growth as invasive lobular carcinoma, yet neoplastic cells have marked nuclear atypia and pleomorphism. Pleomorphic lobular carcinoma and the in situ counterpart, pleomorphic lobular carcinoma in situ, are frequently hormone receptor (estrogen-, progesterone- and androgen-receptor)-positive and E-cadherin and beta-catenin-negative, attaining to their lobular nature. Tumors can also be positive for HER2, p53, ki67 and GCDFP-15 and harbor frequent chromosomal alterations involving gains on 1q and 16p, losses on 11q and 16q, and genomic amplifications in the region of 8q24, 11q13, 12q13, 17q12 and 20q13. Recent gene expression profiling classified pleomorphic lobular carcinoma as 'molecular apocrine' tumors reflecting the frequent apocrine differentiation of the tumors. In support for the aggressive biological features described for pleomorphic lobular carcinoma, accumulating clinical data demonstrate that it has an aggressive clinical course. It is now important to define the most appropriate management strategy for patients diagnosed with pleomorphic lobular carcinoma. PMID- 19284382 TI - Tumor suppressor genes in myeloid differentiation and leukemogenesis. AB - Tumor suppressor genes, such as p53, RB, the INK4-ARF family and PML, suppress malignant transformation by regulating cell cycle progression, ensuring the fidelity of DNA replication and chromosomal segregation, or by inducing apoptosis in response to potentially deleterious events. In myeloid leukemia, hematopoietic differentiation resulting from highly coordinated, stage-wise expression of myeloid transcription and soluble signaling factors is disrupted leading to a block in terminal differentiation and uncontrolled proliferation. This virtually always involves functional inactivation or genetic disruption of one or several tumor suppressor genes in order to circumvent their checkpoint control and apoptosis-inducing functions. Hence, reactivation of tumor suppressor gene function has therapeutic potential and can possibly enhance conventional cytotoxic chemotherapy. In this review, we focus on the role of different tumor suppressor genes in myeloid differentiation and leukemogenesis, and discuss implications for therapy. PMID- 19284385 TI - Biochemical characterization of GSK1070916, a potent and selective inhibitor of Aurora B and Aurora C kinases with an extremely long residence time1. AB - The Aurora kinases AurA, B and C are serine/threonine protein kinases that play essential roles in mitosis and cytokinesis. Among them, AurB is required for maintaining proper chromosome alignment, separation and segregation during mitosis, and regulating a number of critical processes involved in cytokinesis. AurB overexpression has been observed in a variety of cancer cell lines, and inhibition of AurB has been shown to induce tumour regression in mouse xenograft models. In the present study we report the enzymatic characterization of a potent and selective AurB/AurC inhibitor. GSK1070916 is a reversible and ATP-competitive inhibitor of the AurB-INCENP (inner centromere protein) enzyme. It selectively inhibits AurB-INCENP (K(i)*=0.38+/-0.29 nM) and AurC-INCENP (K(i)*=1.5+/-0.4 nM) over AurA-TPX2 (target protein for Xenopus kinesin-like protein 2) (K(i)=490+/-60 nM). Inhibition of AurB-INCENP and AurC-INCENP is time-dependent, with an enzyme inhibitor dissociation half-life of >480 min and 270+/-28 min respectively. The extremely slow rate of dissociation from the AurB and AurC enzymes distinguishes GSK1070916 from two other Aurora inhibitors in the clinic, AZD1152 and VX-680 (also known as MK-0457). PMID- 19284383 TI - The TGF-beta paradox in human cancer: an update. AB - TGF-beta plays an essential role in maintaining tissue homeostasis through its ability to induce cell cycle arrest, differentiation and apoptosis, and to preserve genomic stability. Thus, TGF-beta is a potent anticancer agent that prohibits the uncontrolled proliferation of epithelial, endothelial and hematopoietic cells. Interestingly, tumorigenesis typically elicits aberrations in the TGF-beta signaling pathway that engenders resistance to the cytostatic activities of TGF-beta, thereby enhancing the development and progression of human malignancies. Moreover, these genetic and epigenetic events conspire to convert TGF-beta from a suppressor of tumor formation to a promoter of their growth, invasion and metastasis. The dichotomous nature of TGF-beta during tumorigenesis is known as the 'TGF-beta paradox', which remains the most critical and mysterious question concerning the physiopathological role of this multifunctional cytokine. Here we review recent findings that directly impact our understanding of the TGF-beta paradox and discuss their importance to targeting the oncogenic activities of TGF-beta in developing and progressing neoplasms. PMID- 19284386 TI - Electrospray ionization mass spectrometry as a method for studying the high pressure denaturation of proteins. AB - High-pressure denaturation of proteins can provide important information concerning their folding and function. These studies require expensive and complicated equipment. In this paper, we present a new convenient method for studying high-pressure denaturation of proteins combining DHX (deuterium-hydrogen exchange) and electrospray ionization MS. Application of various values of pressure causes different degrees of protein unfolding resulting in molecules with a different number of protons available for exchange with deuterons. After decompression a protein refolds and a certain number of deuterons are trapped within the hydrophobic core of a refolded protein. Redissolving the deuterated protein in an aqueous buffer initiates the DHX of amides located on the protein surface only, which can be monitored under atmospheric pressure by MS. Depending on the degree of deuteration after high-pressure treatment, the DHX kinetics are different and indicate how many deuterons were trapped in the protein after refolding. The dependence of this number on pressure gives information on the denaturation state of a protein. The distribution of deuterium along the sequence of a high-pressure-denatured protein was studied the ECD (electron-capture induced dissociation) on a Fourier-transform mass spectrometer, enabling the monitoring of high-pressure denaturation with single amino acid resolution. PMID- 19284388 TI - Catheter-related urinary tract infection in patients suffering from spinal cord injuries. AB - Urinary tract infection is commoner in patients with spinal cord injuries because of incomplete bladder emptying and the use of catheters that can result in the introduction of bacteria into the bladder. 145 patients suffering from spinal cord injuries, admitted to the Institute for physical medicine and rehabilitation, Centre for paraplegia of the Clinical Centre of the University of Sarajevo, were included. The patients were divided in three groups according to the method of bladder drainage: Group A (n=61) consisted of patients on clean intermittent catheterization; Group B (n=54) consisted of patients with indwelling catheters; Group C (n=30) consisted of patients who had performed self catheterization. From a total of 4539 urine samples, 3963 (87,3%) were positive and 576 (12,7%) were sterile. More than 90% of the infected patients were asymptomatic. The overall rate of urinary infection amounted to about 2,1 episodes, and bacteriuria to 8,1 episodes per patient. 77% of infections (113/145) were acquired within seven days from catheterization. Infection was usually polymicrobial; the greatest number of urine samples 1770/3943 (44,9%) included more than one bacterium. The vast majority of cases of urinary tract infection and bacteriuria are caused by Gram-negative bacilli and enterococci, commensal organisms of the bowel and perineum, representative of those from the hospital environment. Providencia stuarti (18,9%) being the most common, followed by Proteus mirabilis (16,3%), Escherichia coli (11,8%), Pseudomonas aeruginosa (10,2%), Klebsiella pneumoniae (8,1%), Morganella morgani (5,4%), Acinetobacter baumannii (4,6%), Providencia rettgeri (3,5%). 15,7% of isolates were Gram positive with Enterococcus faecalis (8,6%) as the most common. 55,3% of isolates were multidrug-resistant, and the highest rates of resistance were found among Acinetobacter baumannii (87,8%), Providencia rettgeri (86,7%), Pseudomonas aeruginosa (85,4%), Providencia stuarti (84,3%) and Morganella morgani (81,0%). Lower rates of resistance were found in Group C, i.e. patients on intermittent self-catheterisation. Eradication of organisms was achieved in only 53 (10,05%) of patients; hence, antibiotic therapy had no or very low effect. Significant correlations were found between the method of catheterization and the frequency of bacteriuria and urinary tract infections. The analysis of Group C showed a rate of lower urinary tract infection and bacteriuria than the other two Groups of patients. The objective of this study is the update of etiology and antimicrobial susceptibility in urinary tract infections in this group of patients. In addition, possible correlations between UTI and the type of bladder management were examined. PMID- 19284389 TI - Molecular analysis: microsatellity instability and loss of heterozygosity of tumor suppressor gene in hereditary non-polyposis colorectal cancers (HNPCC). AB - HNPCC (Hereditary non-polyposis colorectal cancers) development is caused by mutation of genes included in system of mismatch repair genes. The mutation exists at 60% of patients in hMSH2 gene, 30% in hMLH1 and 10% both in hPMS1and hPMS2 genes. RER+ exists in about 90% in hereditary non-polyposis colorectal cancer and about 15-28% in sporadic cancers. The purpose of the study was to determine highly sensitive microsatellite markers which can be fast and efficient way of microsatellite screening for detection of HNPCC patients. Moreover, we have analysed the loss of heterozygosity of tumour suppressor genes which could have the diagnostic value in detection of HPNCC patients. PMID- 19284390 TI - Colorectal cancer: prognostic values. AB - After lung cancer colorectal cancer (Cc) is ranked the second, as a cause of cancer-related death. The purpose of this study was to analyze the Cc cases in our material with respect to all prognostic values including histological type and grade, vascular invasion, perineural invasion, and tumor border features. There were investigated 149 cases of resection specimen with colorectal cancer, which were fixed in buffered neutral formalin and embedded in paraffin. Tissue sections (4(mum thick) were cut and stained with H&E. Adenocarcinoma was the most frequent histological type found in 85,90% of cases, in 60,94% of males and 39,06% of females; squamous cell carcinoma in 7,38%, in 63,63% of males and 36,36% of females; mucinous carcinoma in 4,68%, in 57,15% of males and 42,85% of females; while adenosquamous carcinoma, undifferentiated carcinoma and carcinoma in situ in 0,71% of cases each. Dukes' classification was used in order to define the depth of invasion. Dukes B was found in 68,45% of cases, whereas in 31,54% of cases Dukes C was found. As far as histological grading is concerned, Cc was mostly with moderate differentiation (75,16%) with neither vascular nor perineural invasion. Resection margins were in all cases free of tumor. Our data indicate that the pathologic features of the resection specimen constitute the most powerful predictors of postoperative outcome in Cc. Dukes' stage and degree of differentiation provide independent prognostic information in Cc. However, differentiation should be assessed by the worst pattern. PMID- 19284391 TI - Efficacy of tennis elbow (epicondylitis humeri radialis) treatment in CBR "Praxis". AB - Tennis elbow (Epicondylitis humeri radialis) is the most frequent reason that patients with elbow pain report to a physician. The exact cause of elbow pain is still unclear. However, it is certainly causally connected with connective intersection between forearm muscle and bone in the elbow region. In this paper, we analyzed patients that, over the period of 15 years, reported to "Praxis" clinic for elbow pain treatment (Epicondylitis humeri radialis). Of the total number of 228 patients, 126 were male, 101 female while one patient was younger than 14. Initial analysis established that average overall health condition grade was 2,87 at the onset of treatment. Following the treatment completion that grade was 4,48. Of the total number of 223 patients who were treated by combined method of manipulation and local corticosteroid instillation, eight patients received physical therapy as well. Thus, surgical treatment was not necessary in any patient. The patients' treatment included: 1. Application of manipulative methods in order to reestablish mobility in the "blocked" radio-humeral and the upper radio-ulnar joints. 2. Local instillation of corticosteroid depot in order to control inflammation (enthesitis) and thus, eliminate pain and establish physiological conditions for functioning of joint and local structures. Unlike conservative method which includes initial immobilization due to irritation and inflammation development prevention with concomitant analgesic and antirheumatic therapy, initial application of manipulation with reinstatement of joint mobility instead of immobilization with subsequent instillation of steroid preparations achieves functional restitution and fast reinstatement of full working ability, as a rule. PMID- 19284392 TI - Identification and isolation of pharmacologically active triterpenes in Betuale cortex, Betula pendula Roth., Betulaceae. AB - Betulae cortex, Betula pendula Roth., Betulaceae, comprise triterpene substances which are confirmed to posses very important pharmacological activities such as anti-inflammatory, anticancer and antiviral. In this study, extraction of triterpene substances from both, inner and external birch bark was carried out and after that qualitative analysis on betulin, betulinic acid, oleanolic acid and lupeol was performed by method of thin layer chromatography. By this separation method, applying system for development benzene-ethyl acetate-formic acid (36:12:5), is gained a good separation of examined triterpene substances from methanol extracts of inner and external birch bark as well as used standards. From obtained row triterpene mixtures, certain triterpene substances are isolated using method of dry column chromatography. To those substances infrared (IR) spectra were recorded and compared with IR spectra of adequate standards. The study encloses all obtained IR spectra and interpretations on the basis of which can be concluded that triterpene substances, betulin, betulin acid and lupeol isolated from external birch bark give identical characteristic signals and absorbance as referent standards. Method of dry column chromatography has resulted as simple, efficient, repeatable and economical for laboratory conditions. Beside this, a sufficient quantity of examined triterpene substances is also obtained for continuation of their further analytical analysis. PMID- 19284393 TI - Sandfly - Pappataci fever in Bosnia and Herzegovina: the new-old disease. AB - Sandfly fever viruses (SFV) are endemic in the Mediterranean, Middle East, northern African and western Asian countries. Toscana virus (TOSV), serotype of Sandfly fever Naples virus, is among of the three most prevalent viruses associated with meningitis during the warm seasons in northern Mediterranean countries. The historical data of the sandfly fever (Pappataci fever) indicates its origin in Bosnia and Herzegovina at the end of 19th century. There is a long period of time for which there are no data on research related to the SFV in Bosnia and Herzegovina. The purpose of the study was to investigate the presence of sandfly fever in Bosnia and Herzegovina in recent years. The 68 of serum samples were obtained from February 2006 until September 2008 from a group of patients with febrile illness of unknown etiology. The sera were tested on the presence of IgG and IgM antibodies against TOSV by specific serology test- recomLine Bunyavirus IgG/IgM immuno-line assay. The recent TOSV-infection was confirmed in the patients in each year during the study: 10,71% (3/28) in 2008; 9,38% (3/32) in 2007 and 12,50% (1/8) in 2006. The presence of specific antibodies to TOSV in the sera of the patients in recent years indicates re emerging character of the disease in this region. It would be necessary to make biological, epidemiological and clinical research on the TOSV and related phleboviruses to elucidate the problem of SFV in Bosnia and Herzegovina. PMID- 19284394 TI - Effects of aggressive approach to the multiple risk factors for diabetic nephro pathy on proteinuria reduction in diabetes type 2 patients. AB - Dietary interventions with protein and salt restriction, good glucose control, smoking cessation, aggressive blood pressure control, good control of cholesterol and triglycerides, use of ACE inhibitors and ARBs can delay the progression of diabetic nephropathy. The aim of this study was to present the effects of aggressive treatment of the multiple risk factors for diabetic nephropathy on proteinuria in patients with type 2 diabetes. In this study we included 15 patients with diabetes type 2 and insufficient regulation of glycaemia. The patients were followed for three months period. Glycated haemoglobin (HbA1c), fasting plasma glucose (FPG), postprandial plasma glucose (PPG), blood pressure, cholesterol and triglycerides and proteinuria were followed prior and after the study. Prior the study patients were treated with premix insulin divided in two daily doses + metformin after the lunch and they had insufficient regulation of glycaemia. During the study patients were treated with one daily dose of basal insulin, three doses of metformin (2550 mg), one daily dose of atorvastatin (20 mg) and one daily dose of ramipril (5 to 10 mg). Doses of insulin were titrated separately for each patients (0,7-1,0 IU/kg). Patients were advised to start with lifestyle modification, increased physical activity and dietary interventions with protein and salt restriction, energy restricted diet and smoking cessation. A total of 20 patients (male 12 and female 8) with diabetes type 2 were studied. The mean age of the subjects was 53+/-5,25 years. The mean diabetes duration was 4,05+/-1,96 years. The mean body mass index decreased from 28,1+/-1,67 kg/m2 to 25,9 +/-1,22 kg/m2 after the study. Mean HbA1c decreased from 8,82 +/- 0,53 % to 7,15 +/- 0,23 % (p<0,05). Mean fasting glycemia decreased from 8,79+/-0,58 mmol/dm3 to 7,03+/-0,18 mmol/dm3 (p < 0,05). Mean postmeal glycemia decreased from 9,93 +/- 0,77 mmol/dm3 to 7,62 +/- 0,42 mmol/dm3 (p<0,05). The mean cholesterol level decreased from 7,99 +/-0,64 mmol/dm3 to 5,93 +/- 0,65 mmol/dm3 (p<0,05). The mean triglicerides level decreased from 4,05 +/- 0,97 mmol/dm3 to 1,96 +/- 0,24 mmol/dm3 (p<0,05). The significant decrease of proteinuria was recorded, prior the study the mean albuminuria was 1,05 +/- 0,31 g/dm3 and after the study was 0,07 +/- 0,145 g/dm3 (p<0,05). Mean blood pressure prior the study was 153+/-8,69/91,5 +/- 3,78 mm Hg (p<0,05), after the study was 125 +/- 6,32/ 79,25+/-3,26 mmHg. Effective control of glycaemia, blood pressure, cholesterol and triglycerides, use of ACE inhibitors, dietary interventions with protein and salt restriction, smoking cessation, can delay the progression of nephropathy in type 2 diabetes. PMID- 19284395 TI - Effects of functional electrical stimulation in rehabilitation with hemiparesis patients. AB - Cerebrovascular accident is a focal neurological deficiency occurring suddenly and lasting for more than 24 hours. The purpose of our work is to determine the role of the functional electrical simulation (FES) in the rehabilitation of patients with hemiparesis, which occurred as a consequence of a cerebrovascular accident. This study includes the analysis of two groups of 40 patients with hemiparesis (20 patients with deep hemiparesis and 20 patients with light hemiparesis), a control group which was only treated with kinesiotherapy and a tested group which was treated with kinesiotherapy and functional electrical stimulation. Both groups of patients were analyzed in respect to their sex and age. Additional analysis of the walking function was completed in accordance with the BI and RAP index. The analysis of the basic demographical data demonstrated that there is no significant difference between the control and tested group. The patients of both groups are equal in respect of age and sex. After 4 weeks of rehabilitation of patients with deep and light hemiparesis there were no statistically significant differences between the groups after evaluation by the BI index. However, a statistically significant difference was noted between the groups by the RAP index among patients with deep hemiparesis. After 8 weeks of rehabilitation the group of patients who were treated with kinesiotherapy and functional electrical stimulation showed better statistically significant results of rehabilitation in respect to the control group with both the BI index and the RAP index (p<0,001). In conclusion, we can state that the patients in rehabilitation after a cerebrovascular accident require rehabilitation longer than 4 weeks. Walking rehabilitation after stroke is faster and more successful if we used functional electrical stimulation, in combination with kinesiotherapy, in patients with disabled extremities. PMID- 19284396 TI - Survival benefit of the late percutaneous coronary intervention in the patients after acute myocardial infarction who are or who are not treated with thrombolysis. AB - The impact of late percutaneous coronary intervention (PCI) in the patients after acute myocardial infarction (AMI) on long term mortality remains to be established. At currently, thrombolysis is accepted as standard therapy when PCI is not immediately available. However, PCI is often performed in stable patients with AMI who are/are not received thrombolysis . We performed the trial that enrolled myocardial infarction patients treated with thrombolysis, late PCI and medically to assess the potential benefits of delayed PCI. We follow up 164 consecutive patients after AMI one year. The patients are divided in two groups; first group-66 patients who received reperfusion (37 patients received only thrombolysis, 10 patients received thrombolysis and PCI 7-9 days after thrombolysis and 19 patients underwent only PCI after 7-9 days) and second group 98 patients medically treated. One year mortality was 3% in the reperfusion group (2/66) and 14,3% in the medical group (14/98) (p=0,016). There were not significant differences between groups about other end points-reinfarctus, coronary artery bypass surgery and PCI performed later after discharge. The major predictors of one year mortality were ages (p<0,001) and ejection fraction (p=0,003). Also, therapy with beta-blockers (p=0,002), statins (p=0,001) and ACE inhibitors (p=0,024) was associated with better survival. Delayed PCI performed 7 9 days after AMI in the patients who underwent thrombolysis or those did not improves outcome at long-term follow-up. PMID- 19284397 TI - Laser therapy of painful shoulder and shoulder-hand syndrome in treatment of patients after the stroke. AB - The common complication after stroke is pain and dysfunction of shoulder of paralyzed arm, as well as the swelling of the hand. The aim of this study was to determine the effects of LASER therapy and to correlate with electrotherapy (TENS, stabile galvanization) in subjects after stroke. We analyzed 70 subjects after stroke with pain in shoulder and oedema of paralyzed hand. The examinees were divided in two groups of 35, and they were treated in the Clinic for Physical Medicine and Rehabilitation in Tuzla during 2006 and 2007. Experimental group (EG) had a treatment with LASER, while the control group (CG) was treated with electrotherapy. Both groups had kinesis therapy and ice massage. All patients were examined on the admission and discharge by using the VAS, DASH, Barthel index and FIM. The pain intensity in shoulder was significantly reduced in EG (p<0,0001), swelling is lowered in EG (p=0,01). Barthel index in both groups was significant higher (p<0,01). DASH was significantly improved after LASER therapy in EG (p<0,01). EG had higher level of independency (p<0,01). LASER therapy used on EG shows significantly better results in reducing pain, swelling, disability and improvement of independency. PMID- 19284398 TI - Surveillance of intrahospital infections at the clinic for gynaecology and obstetrics. AB - Intrahospital infections (IHI) and antibiotics resistance are the problems which exist in virtually all hospitals in the world. The main aim of the present research is establishing of epidemiological surveillance over occurrence of IHI at the Clinic for Gynaecology and Obstetrics at the University Clinical Center Tuzla and thus identifies: types of bacteria which cause IHI, types of infection according to anatomical localization and research resistance organisms causing of IHI on antimicrobial drugs. A study was implemented on all patients admitted to Clinic for Gynaecology and Obstetrics during the period of one year and who subsequently developed infection. Determination of intrahospital infections was done according to criteria defined by the Centres for Disease Control and Prevention from the United States. The results of our work have shown that both urinary tract infections and surgical site infections are the most frequent. As IHI causers the most found are gram-negative organisms (73,7%), such as Escherichia coli (29,8%), right after that Klebsiella pneumoniae (24,6%), Pseudomonas aeruginosa (14%) and Proteus mirabilis (5,3%) (p<0,05). Gram-positive organisms as causers of IHI are registered in 26,3% cases. Out of that Streptococcus species are isolated in 10,5% cases, Staphylococcus aureus (8,8%) and coagulasa negative staphylococci (7%) (p>0,05). High percent resistance of bacteria was evident to beta-lactams, aminoglycosids and cephalosporin's of third generation. Gram-positive organisms were 100% sensitive to vancomycin, while gram negative organisms manifested the high percent of sensibility to imipenem and cefepime. PMID- 19284399 TI - The statins in prevention of coronary heart diseases in type 2 diabetics. AB - We aimed to determine whether the administration of statins to type 2 diabetics without pre-existing CHD reduced the incidence of CHD and their effects on cholesterol and CRP levels. All the participants were carefully interviewed, clinically examined, and laboratory tested to exclude conditions likely to provoke an inflammatory response that was an exclusion criterion. EXCLUSION CRITERIA: Serious heart, liver or kidney problems, history of renal transplant, recent history of drug or alcohol abuse, HbA1c>10%, blood pressure >140/90 mmHg, BMI >35 kg/m2, triglycerides >3,0 mmol/dm3. 95 obese diabetics (mean age 60,9 years and BMI=31,59 kg/m2, diabetes duration more than 10 years) without pre existing CHD, were included in the analysis and were randomized to simvastatin (25 female and 20 male used 40 mg simvastatin daily) or placebo (30 female and 20 male) group. After six months, simvastatin significantly lowered CRP levels by 19%, (p<0,01), cholesterol levels by 18%, TG levels by 8%, LDL levels by 20% and VLDL levels by 17%, whereas there was no change with placebo. After one year the difference sustained between groups. Coronary events were rarely in the simvastatin group (6,6%) than in the placebo group (14%). Coronary revascularizations were 4 in the placebo group and 1 in the simvastatin group. Rate of stroke was more often in the placebo group (18%) than in the simvastatin group (8,8%). So, reduction of acute CHD events is for 7,4% in the simvastatin group. Positive correlation was between CRP and CVD (r=0,29). Statin therapy reduced the risk of coronary heart disease in diabetics without CHD. PMID- 19284400 TI - Early detection of melanoma skin cancer. AB - Primary skin melanoma and skin cancers have been more prevalent in the previous decades and therefore have become a very significant public health problem. Dermatologists of the Skin and Venereal Diseases Clinic of the University of Sarajevo Clinics Centre initiated the first public preventive action called "Days of Fighting Melanoma" in May 2008. The objective of the campaign was to provide free dermatological examinations to all volunteers and to inform, through the media, wider population on early signs and recognition of skin cancer and the importance of sun protection. The total of 325 citizens were examined clinically and with dermatoscope in the period between 5 and 31 May 2008. The examination also included histological diagnoses: 7 patients with confirmed melanoma, 30 with basal cell carcinoma and 2 with spinocellular carcinoma. The results suggested a need for the expansion of the campaign to other towns in our country in order to demonstrate the importance of early detection of the disease and treatment options. PMID- 19284401 TI - Characteristics of symptomatic epilepsy in patients with brain tumours. AB - The aim of our work is to determine the total number, age, gender of the patients with the symptomatic epileptic seizures associated with brain tumours, tumour location, clinical signs and characteristics of epileptic seizures. We have analyzed medical documentation of the patients with brain tumours hospitalized at the Department of Neurology, University of Sarajevo Clinics Centre. This study is retrospective and includes time period from 1st January 2000 until 31st December 2005. During the observed period at the Department of Neurology in Sarajevo there were in total 9753 hospitalized patients, from which 101 (1,1%) patients with the brain tumour diagnosis. Average patient's age was 62,60 +/- 1,28 years. In one third of the patients (32%) were recorded epileptic seizures, without significant difference between genders. In case of symptomatic epilepsy, significantly more frequent locations of tumours were: in several lobes (28%), parietal lobe (25%), as well as frontal and temporal lobe (18,8% each), while there were no changes in cerebellum and brain stem (chi2 =7,174, p<0,05). The most prominent signs of illness in our sample were hemiparesis with the cranial nerves lesion (56,3%), speech problems (25%). Normal neurologic findings were significantly more frequent among patients with the symptomatic epilepsy (chi2 =6,349, p<0,05). The most often was a single seizure (59%), in 38% of cases there were recorded series of seizures, and only 3% of patients had status epilepticus. In relation to the type of seizures, the most often are simple partial seizures with or without secondary generalization (66%), than generalized convulsive (31%), and the rarest one are complex partial seizures (3%). Symptomatic epilepsy in case of brain tumours occurs in one third of patients, at older age, and in both genders. The lesion usually affects several lobes and cause simple partial seizures with or without secondary generalization. The most often clinical signs in case of all brain tumours are cranial nerves lesion and hemiparesis, while the normal neurologic findings are significantly dominant in the group of patients with the epileptic seizures. PMID- 19284402 TI - Primary open-angle glaucoma and serum lipids. AB - The aim of this paper was to prove the relation between serum lipid values (cholesterol, triglyceride, low density cholesterol, high density cholesterol and primary open -angle glaucoma. The study includes two patient groups: 50 patients with primary open-angle glaucoma and 50 patients without this disease. However, all 100 patients were tested for serum lipid values. The research covered a period of six months (from May to December 2007.). Primary open-angle glaucoma was diagnosed with intraocular pressure values (between 20,1 and 25,6 mm Hg) measured with Schiotz tonometry. The visual field changes were confirmed with Goldmann perimetry. The gonioscopies were done for diagnosis confirmation. The serum lipid values were confirmed with enzymatic colorimetry in vitro method. U test (Mann-Witney-Wilcox test) and t-test, as nonparametric tests, were used for statistical evaluations. The cholesterol mean value in the test group was 6,14 mol/dm (3) (3,20-8,10 mol/dm (3)) whereas in the control group it was 5,96 mol/dm(3) (2,70-8,80 mol/dm(3)). U-test was with negative ranks (z=-0,83 AS=0,678). The triglyceride mean value in the test group was 2,38 mol/dm (3) (0,84-11,73 mol/m (3)) and in the control group it was 2,04 mol/dm (3) (0,63-5,89 mol/dm(3)). U-test was with positive ranks (z=0,950 AS=0,342). High density cholesterol was average in the test group with 1,45 mol/dm (3) (0,71-3,40 mol/dm (3)) and in the control group 1,40 mol/m (3) (0,80-3,20 mol/dm (3)). Low density cholesterol in the test group was 3,98 mol/m (3) (1,82-6,49 mol/m (3)) and in the control group 4,08 mol/m (3) (2,69-5,69 mol/m (3)). These results had positive ranks according to U-test. Serum lipid values could be one of predictable factors in primary open-angle glaucoma diagnosis. Due to the patient age, cholesterol values, as common factors in primary open-angle glaucoma and atherosclerosis genesis, could be concern in the same aetiology based on dyslipidaemia as well. PMID- 19284403 TI - Influence of type and neutralisation capacity of antacids on dissolution rate of ciprofloxacin and moxifloxacin from tablets. AB - Dissolution rate of two fluoroquinolone antibiotics (ciprofloxacin and moxifloxacin) was analysed in presence/absence of three antacid formulations. Disintegration time and neutralisation capacity of antacid tablets were also checked. Variation in disintegration time indicated the importance of this parameter, and allowed evaluation of the influence of postponed antacid fluoroquinolone contact. The results obtained in this study showed decreased dissolution rate of fluoroquinolone antibiotics from tablets in simultaneous presence of antacids, regardless of their type and neutralisation capacity. PMID- 19284404 TI - Evaluation of renal function in children with mitochondrial respiratory chain complex defect: usefulness of cystatin C. AB - AIM: Mitochondrial disorders with respiratory chain complex defect are known to show a broad spectrum of clinical phenotypes, though their contribution to renal disease has not received much attention. The aim of this study was to evaluate renal function using different markers of the glomerular filtration rate (GFR) in children with mitochondrial respiratory chain complex (MRC) defects. METHODS: Twenty-two patients with MRC I defect confirmed by biochemical enzyme assays using muscle tissue, were evaluated for renal function with diethylenetriaminepentaacetic acid (DTPA) renal scintigraphy, urine creatinine clearance, serum creatinine (Scr) and cystatin C (Cys C) levels. RESULTS: All patients showed lower levels within normal range of Scr (0.4-0.7 mg/dL, mean 0.47 mg/dL), and with no significant correlation with DTPA-GFR. Cys C showed significant correlation with DTPA-GFR (p < 0.01) providing higher diagnostic accuracy than creatinine in patients with MRC defect. CONCLUSION: In mitochondrial disorder with MRC defect, Scr could not fully reflect renal function due to the relatively small body mass of patients. Considering the limited clinical availability of DTPA renal scintigraphy, Cys C should be taken as the first step to evaluate GFR in mitochondrial disorders. PMID- 19284405 TI - The unbearable cost of severe asthma in underprivileged populations. PMID- 19284406 TI - Anaphylaxis to amidotrizoate proved by skin testing and flow cytometry-based basophil activation test. PMID- 19284407 TI - Review article: pain and chronic pancreatitis. AB - BACKGROUND: Pain in chronic pancreatitis chronic pancreatitis is a frustrating and challenging symptom for both the patient and clinician. It is the most frequent and most significant symptom. Many patients fail the currently available conservative options and require opiates or endoscopic/surgical therapy. Aim To highlight the pathophysiology and management of chronic pancreatitis pain, with an emphasis on recent developments and future directions. METHODS: Expert review, utilizing in addition a comprehensive search of PubMed utilizing the search terms chronic pancreatitis and pain, treatment or management and a manual search of recent conference abstracts for articles describing pain and chronic pancreatitis. RESULTS: Pancreatic pain is heterogenous in its manifestations and pathophysiology. First-line medical options include abstinence from alcohol and tobacco, pancreatic enzymes, adjunctive agents, antioxidants, and non-opiate or low potency opiate analgesics. Failure of these options is not unusual. More potent opiates, neurolysis and endoscopic and surgical options can be considered in selected patients, but this requires appropriate expertise. New and better options are needed. Future options could include new types of pancreatic enzymes, novel antinociceptive agents nerve growth factors, mast cell-directed therapy, treatments to limit fibrinogenesis and therapies directed at the central component of pain. CONCLUSIONS: Chronic pancreatitis pain remains difficult to treat. An approach utilizing conservative medical therapies is appropriate, with more invasive therapies reserved for failure of this conservative approach. Treatment options will continue to improve with new and novel therapies on the horizon. PMID- 19284409 TI - Poly I:C-induced expression of intercellular adhesion molecule-1 in intestinal epithelial cells. AB - Intercellular adhesion molecul-1 (ICAM-1) is a transmembrane glycoprotein belonging to the immunoglobulin superfamily of adhesion molecules and plays perdominant roles in recruitment and trafficking of leucocytes to sites of inflammation. ICAM-1 expression in intestinal epithelial cells (IECs) is enhanced by several stimuli, such as proinflammatory cytokines, bacterial infections or pathogen-associated molecular patterns. One of these stimuli, double-stranded RNA (dsRNA), is a by-product of viral replication and can be recognized by its cognate receptor Toll-like receptor 3 (TLR-3). In spite of expression of both TLR 3 and ICAM-1 in IECs, correlation between TLR-3-signalling and ICAM-1 expression has never been examined in IECs. In the present study, we investigated whether poly I:C, an analogue of dsRNA, can stimulate the expression of ICAM-1 in IEC line, HT-29. Poly I:C-stimulation up-regulated the expression of ICAM-1 mRNA by real-time polymerase chain reaction. Enhanced expression of ICAM-1 was confirmed in protein level by immunofluoresense cell staining and enzyme-linked immunosorbent assay by measuring the released soluble ICAM-1 in culture supernatant. As the stimulation effect was reduced by pre-treatment of the cells with anti-TLR-3 antibody, poly I:C-binding signal was thought to be sensed by TLR 3 on the surface of HT-29. The results of luciferase assay and nuclear factor kappa-b (NF-kappaB) inhibitor treatment experiments indicated that the downstream signal was mainly transduced by transcription factor, NF-kappaB. All these results demonstrated the connection between TLR-3 signalling and ICAM-1 expression in HT-29 cells and indicated the importance of coordinated function of both innate and adaptive immunity against viral infections. PMID- 19284411 TI - Abstracts of Diabetes UK's Annual Professional Conference 2009. March 11-13, 2009. Glasgow, United Kingdom. PMID- 19284410 TI - Signet ring cells in fine needle aspiration cytology of breast carcinomas: review of the cytological findings in ten cases identified by histology. AB - OBJECTIVE: To establish whether the presence of signet ring cells (SRCs) in histology sections of breast carcinoma cases was reflected by their presence in fine needle aspiration cytology (FNAC) smears, correlating to the histological type of breast carcinoma. METHODS: We reviewed the FNAC findings of ten cases that had been diagnosed as primary breast carcinoma with SRCs on histological sections between 1998 and 2007. Slides and histological sections were obtained from the archives of Ege University Hospital. RESULTS: FNA smears were reviewed for the following cytomorphological features: background, cellularity, architecture, nuclear pleomorphism and the presence of SRCs. The background was bloody in eight cases, necrotic in one, and clean in one. There was no mucinous material in any of the cases. Cellularity was prominent in five cases (hypercellular), moderate in three (cellular) and low in two (hypocellular). Loosely cohesive groups of tumour cells of varying size were observed in all cases. A plasmacytoid appearance to some of the tumour cells was seen in all cases and discohesive tumour cells were present in eight. Nuclear pleomorphism was high in six cases and moderate in four. SRCs were observed in seven of the ten cases. Two of these seven cases also had a tubular pattern and one had tumour giant cells. CONCLUSIONS: FNAC should be evaluated carefully regarding the presence of SRCs when cells with a plasmacytoid appearance are observed in either hyper- or hypocellular smears. The presence of single SRCs in FNACs with hypercellularity, high nuclear grade and tubular formation or tumour giant cells may be a clue in favour of ductal carcinoma. The presence of single SRCs in FNACs with hypocellularity and mild to moderate nuclear grade may be suggestive of lobular carcinoma. However, larger studies would be needed to establish the predictive value of the presence of SRCs on FNAC. PMID- 19284414 TI - Stabilization of an archaeal DNA-sliding clamp protein, PCNA, by proteasome activating nucleotidase gene knockout in Haloferax volcanii. AB - Many details of structure, function and substrate specificity of eukaryotic proteasomal systems have been elucidated. This information far-exceeds that available for the archaeal and bacterial counterparts. While structural and functional studies have provided some insight into the workings of prokaryotic proteasomes, the question of substrate targeting and global cellular influence remain largely unaddressed. In this communication, we report an over 720-fold increase in the half-life of the DNA-sliding clamp protein proliferating cell nuclear antigen after knockout of the panA gene, encoding a proteasome-activating nucleotidase A, on the chromosome of the halophilic archaeon Haloferax volcanii. This discovery marks the first identification of a protein stabilized by an archaeal proteasome mutation and provides a starting point for investigations into substrate recognition mechanisms. The findings also begin to address the functional role of proteasomal systems within the scope of the archaeal cell. PMID- 19284415 TI - Fetal blood analysis is not a primary method in prenatal diagnosis of thalassemia. PMID- 19284416 TI - Depletion of L-ascorbic acid alternating with its supplementation in the treatment of patients with acute myeloid leukemia or myelodysplastic syndromes. AB - PURPOSE: L-ascorbic acid (LAA) modifies the in vitro growth of leukemic cells from approximately 50% of patients with acute myeloid leukemia (AML) or myelodysplastic syndromes (MDS). To test the hypothesis that depletion of LAA, alternating with supplementation to prevent scurvy, would provide therapeutic benefit, a single-arm pilot trial was conducted (ClinicalTrials.gov identifier: NCT00329498). Experimental results: During depletion phase, patients with refractory AML or MDS were placed on a diet deficient in LAA; during supplementation phase, patients received daily intravenous administration of LAA. An in vitro assay was performed pretherapy for LAA sensitivity of leukemic cells from individual patients. RESULTS: Of 18 patients enrolled, eight of 16 evaluable patients demonstrated a clinical response. Responses were obtained during depletion (four patients) as well as during supplementation (five patients) but at a pharmacologic plasma level achievable only with intravenous administration. Of nine patients for whom the in vitro assay indicated their leukemic cells were sensitive to LAA, seven exhibited a clinical response; compared with none of six patients who were insensitive to LAA. CONCLUSIONS: The clinical benefit, along with a conspicuous absence of significant adverse events, suggests that further testing of LAA depletion alternating with pharmacologic dose intravenous supplementation in patients with these and other malignancies is warranted. PMID- 19284417 TI - Solitary plasmocytoma: improvement in critical organs sparing by means of helical tomotherapy. AB - PURPOSE: Helical tomotherapy (HT) was assessed in two patients with paramedullar solitary bone plasmocytoma. We compared doses delivered to critical organs, according HT plan or tridimensional conformal plan. METHODS AND MATERIALS: One male (patient no. 1), 67 yr-old and one female (patient no. 2), 37-yr-old, with histologically, biologically and radiological confirmed paramedullar solitary plasmocytoma have been treated in our department between November 2007 and February 2008 using HT. The prescription dose was 40 Gy in 20 fractions. This HT treatment planning was compared with a routine dosimetric work that was executed for a standard conformal radiotherapy treatment planning. RESULTS: Treatment tolerance was excellent, without any side effects. Both patients achieved 9-month complete remission. HT resulted in substantial critical organs sparing. For patient no. 1, dose delivered to 20% of the total intestine volume was reduced from 28 Gy for conformal radiotherapy to 13 Gy for HT. Radiation dose delivered to 20% of the left kidney was reduced from 25 Gy to 7 Gy. For patient no. 2, volume of left lung that received at least 20 Gy was 12% for conformal radiotherapy vs. 6% for HT. CONCLUSIONS: For paramedullar solitary plasmocytoma, HT has the potential to significantly improve the quality of the dose distribution both in terms of better dose homogeneity within the planning target volume and more efficient sparing of critical organs. PMID- 19284418 TI - Health related quality of life in a nationally representative sample of haematological patients. AB - OBJECTIVES: Knowledge of health related quality of life of haematological patients is limited. This study aimed at investigating the prevalence and predictors of symptoms and problems in a representative sample of haematological patients in Denmark. METHODS: A random sample of patients with leukaemia, multiple myeloma and advanced lymphoma (n = 732) received the European Organisation for Research and Treatment of Cancer quality-of-life questionnaire (EORTC QLQ-C30). Mean scores were calculated. In addition, scores were dichotomised using two thresholds: patients reporting at least 'a little' of each EORTC QLQ-C30 symptom/problem were classified as having a 'symptom/problem', and patients reporting at least 'quite a bit' were classified as having a 'severe symptom/problem'. Multiple logistic regression was used to identify predictors. RESULTS: In total, 470 (64%) patients participated. The most frequent symptoms/problems were fatigue (55%; severe 20%), reduced role function (49%; severe 23%), insomnia (46%; severe 15%), and pain (37%; severe 15%). Older patients and patients in active antineoplastic treatment had more symptoms and problems. There was only little evidence of social inequalities. CONCLUSION: This is probably the first nationally representative study of symptoms and problems in haematological patients. These patients have symptoms/problems that deserve attention. Health related quality of life is an important issue in haematological malignancies. PMID- 19284419 TI - Infection prevention strategies in a stem cell transplant unit: impact of change of care in isolation practice and routine use of high dose intravenous immunoglobulins on infectious complications and transplant related mortality. AB - OBJECTIVE: Nursing in 'live islands' and routine high dose intravenous immunoglobulins after allogeneic hematopoietic stem cell transplantation were abandoned by many teams in view of limited evidence and high costs. METHODS: This retrospective single-center study examines the impact of change from nursing in 'live islands' to care in single rooms (SR) and from high dose to targeted intravenous immunoglobulins (IVIG) on mortality and infection rate of adult patients receiving an allogeneic stem cell or bone marrow transplantation in two steps and three time cohorts (1993-1997, 1997-2000, 2000-2003). RESULTS: Two hundred forty-eight allogeneic hematopoetic stem cell transplantations were performed in 227 patients. Patient characteristics were comparable in the three cohorts for gender, median age, underlying disease, and disease stage, prophylaxis for graft versus host disease (GvHD) and cytomegalovirus constellation. The incidence of infections (78.4%) and infection rates remained stable (rates/1000 days of neutropenia for sepsis 17.61, for pneumonia 6.76). Cumulative incidence of GvHD and transplant-related mortality did not change over time. CONCLUSIONS: Change from nursing in 'live islands' to SR and reduction of high dose to targeted IVIG did not result in increased infection rates or mortality despite an increase in patient age. These results support the current practice. PMID- 19284430 TI - The beginning of the end (of life care strategy). PMID- 19284431 TI - A literature review of the application of the Geriatric Depression Scale, Depression Anxiety Stress Scales and Post-traumatic Stress Disorder Checklist to community nursing cohorts. AB - AIMS AND OBJECTIVES: To explore through literature review the appropriateness of three common tools for use by community nurses to screen war veteran and war widow(er) clients for depression, anxiety and post-traumatic stress disorder. BACKGROUND: War veterans and, to a lesser extent, war widow(er)s, are prone to mental health challenges, especially depression, anxiety and post-traumatic stress disorder. Community nurses do not accurately identify such people with depression and related disorders although they are well positioned to do so. The use of valid and reliable self-report tools is one method of improving nurses' identification of people with actual or potential mental health difficulties for referral to a general practitioner or mental health practitioner for diagnostic assessment and treatment. The Geriatric Depression Scale, Depression Anxiety Stress Scales and Post-traumatic Stress Disorder Checklist are frequently recommended for mental health screening but the appropriateness of using the tools for screening war veteran and war widow(er) community nursing clients who are often aged and have functional impairment, is unknown. DESIGN: Systematic review. CONCLUSIONS: Current literature informs that the Geriatric Depression Scale accurately predicts a diagnosis of depression in community nursing cohorts. The three Depression Anxiety Stress Scales subscales of depression, anxiety and stress are valid; however, no studies were identified that compared the performance of the Depression Anxiety Stress Scales in predicting diagnoses of depression or anxiety. The Post-traumatic Stress Disorder Checklist predicts post traumatic stress disorder in community cohorts although no studies meeting the selection criteria included male participants. RELEVANCE TO CLINICAL PRACTICE: This review provides recommendations for the use of the Geriatric Depression Scale, Depression Anxiety Stress Scales and The Post-traumatic Stress Disorder Checklist based on examination of the published evidence for the application of these screening tools in samples approximated to community nursing cohorts. Findings and recommendations would guide community nurses, managers and health planners in the selection of mental health screening tools to promote holistic community nursing care. PMID- 19284432 TI - Factors associated with the prevalence of violent behaviour among residents living in nursing homes. AB - AIM: The aim of this study was to investigate the associations between environmental and organisational factors as well as resident and caregiver characteristics in nursing home wards with a high respectively low prevalence of residents with violent behaviour. BACKGROUND: Earlier studies have indicated that different factors are related to violent behaviour among residents living in nursing homes, such as environmental and organisational variations, and resident and caregiver characteristics. However, few studies have simultaneously examined the relationship between these factors. DESIGN: A cross-sectional descriptive survey design. METHODS: The study was performed in 10 nursing homes consisting of 33 wards. Data were collected using the Multi-Dimensional Dementia Assessment Scale and the Geriatric Rating Scale. Variables concerning organisation and environment were gathered by means of a questionnaire. Differences between wards with high (HPW) or low prevalence of violence (LPW) were analysed. RESULTS: In HPWs, the prevalence of behaviour and psychiatric symptoms, residents needing assistance with dressing and psychological workload were found to be higher, while job satisfaction was lower compared to LPWs. This study has also shown that caregivers in HPWs had less experience of working with older people and they experienced their working climate as less positive. Furthermore, HPWs had more residents, lower caregiver-to-resident ratio and longer corridors, and caregivers in these wards experienced more difficulties to supervise the residents. CONCLUSION: This study has shown that the prevalence of residents with violent behaviour is significantly associated with other behavioural and psychiatric symptoms and ADL (activities of daily life)-functions, as well as caregivers' experiences of job satisfaction and psychological workload. RELEVANCE TO CLINICAL PRACTICE: This study underlines the importance of a multifactorial approach to understand the prevalence of violent behaviour, including the physical environment, organisational factors, as well as characteristics of the resident and the caregiver. PMID- 19284433 TI - Restraint use among nursing home residents: cross-sectional study and prospective cohort study. AB - AIMS AND OBJECTIVES: To investigate (1) the prevalence of physical restraints and psychoactive medication, (2) newly administered physical restraints, frequency of application of the devices and frequency of psychoactive medication on demand during 12-month follow-up and (3) characteristics associated with restraint use in nursing homes. BACKGROUND: High quality data on restraint use in German nursing homes are lacking so far. Such information is the basis for interventions to achieve a restraint-free care. DESIGN: Cross-sectional study and prospective cohort study. SETTING AND SUBJECTS: Thirty nursing homes with 2367 residents in Hamburg, Germany. METHODS: External investigators obtained prevalence of physical restraints by direct observation on three occasions on one day, psychoactive drugs were extracted from residents' records and prospective data were documented by nurses. RESULTS: Residents' mean age was 86 years, 81% were female. Prevalence of residents with at least one physical restraint was 26.2% [95% confidence interval (CI) 21.3-31.1]. Centre prevalence ranged from 4.4 to 58.9%. Bedrails were most often used (in 24.5% of residents), fixed tables, belts and other restraints were rare. Prevalence of residents with at least one psychoactive drug was 52.4% (95% CI 48.7-56.1). The proportion of residents with at least one physical restraint after the first observation week of 26.3% (21.3-31.3) cumulated to 39.5% (33.3-45.7) at the end of follow-up (10.4 SD 3.3 months). The relative frequency of observation days with at least one device ranged from 4.9 64.8% between centres. No characteristic was found to explain centre differences. CONCLUSIONS: The frequency of physical restraints and psychoactive drugs in German nursing homes is substantial. Pronounced centre variation suggests that standard care is possible without restraints. RELEVANCE TO CLINICAL PRACTICE: Effective restraint minimisation approaches are urgently warranted. An evidence based guideline may overcome centre differences towards a restraint-free nursing home care. PMID- 19284434 TI - Sexual behaviour of institutionalised residents with dementia--a qualitative study. AB - AIMS: The purpose of this study was to explore the characteristics of and the contexts related to sexual behaviours among institutionalised residents with dementia. BACKGROUND: Institutionalised residents with dementia are frequently unable to manage their sexual needs properly resulting in caregivers having a more conservative and passive attitude toward residents with dementia than those with higher cognitive status. DESIGN: A grounded theory study. METHODS: The participants in this study were institutionalised older people with dementia and their formal caregivers. Data were collected using in-depth, face-to-face interviews of 12 formal caregivers and by observing 12 institutional older people with dementia for three days. All observations were recorded and interview data were tape recorded and transcribed verbatim. RESULTS: The results indicated that the predisposing factors included having opportunity, presence of a cooperative target and personal space without privacy. The sexual expressions of institutionalised older people with dementia included: physically intimate touch, sexual expression without touching others and sexual talk. Responses by individuals to sexual behaviour from another resident included neutral response, negative response and positive response. CONCLUSION: To provide better care, it is recommended that an inventory regarding sexual expression for clinical and research usage be constructed from the research results. RELEVANCE TO CLINICAL PRACTICE: The findings of this study can provide the basis to develop on-the-job training programmes for sexual education of residents with dementia in institutions. Regular seminars on sexual care for the residents with dementia might be beneficial for managing sexual issues among residents and to decrease caregivers' burden. PMID- 19284435 TI - The impact of lifestyle on the physical fitness of primary school children. AB - AIMS: To compare the physical fitness levels of Hong Kong and mainland Chinese school children and to study the association between any differences and their respective lifestyles. BACKGROUND: Genes and lifestyle are both factors contributing to health. Hong Kong is a Chinese city with distinctive lifestyle features such as a different diet and different entertainment and education from the rest of China. Using samples of the same ethnicity and investigating differences in the physical fitness parameters between Hong Kong and mainland Chinese school children might reveal the impact of some lifestyle factors on health. DESIGN: Survey. METHOD: Primary school children (n = 522) were recruited. Demographic data were collected by questionnaire. Physical fitness tests were carried out. RESULTS: Significant differences were found in height, body weight, sit and reach, long jump, running 50 m and lung capacity. Hong Kong children were found to be taller and heavier at ages 6 and 7, but heavier with similar height to that of mainland children at ages 8-12. Other results showed better physical fitness on the part of mainland children. DISCUSSION; Although the people of Hong Kong share the same ethnicity as the majority of mainland Chinese, the eastern mixed-with-western diet, education and entertainment of Hong Kong children differ greatly. The findings in terms of physical fitness revealed variations in body build, flexibility, cardiovascular function and muscle power between the two groups. CONCLUSION: Lifestyle could possibly be a key factor and predictor of physical fitness, providing strong evidence to support the interaction between lifestyle and genes in their impact on our health. RELEVANCE TO CLINICAL PRACTICE: Lifestyle could possibly be a key factor and predictor of physical fitness and health outcomes. The finding of this study may enhance the identification of healthy lifestyle which may improve health outcomes in children. PMID- 19284436 TI - Use of non-conventional nurses' attire in a paediatric hospital: a quasi experimental study. AB - AIMS: To test the impact of a multi-coloured non-conventional attire on a population of children admitted to a paediatric hospital. DESIGN: Quasi experimental before-after controlled study. BACKGROUND: It has been suggested that non-conventional nurses' uniforms in paediatric settings may contribute to lowering children's distrust towards healthcare providers and reduce fear. Little formal research has investigated on the impact of nursing attire in a paediatric setting. No study has so far analysed the effects in actual use of a non conventional, other than the traditional type of uniform, on a paediatric hospitalised population. Design: A quasi-experimental study. METHODS: We introduced multi-coloured nurses' attire in two wards of a paediatric hospital. Using open questions and semantic differential scales (SDS), we evaluated the effects of this non-conventional attire on a group of hospitalised children, compared to sex-and-age-matched controls interviewed before the introduction. Parents were also interviewed. RESULTS: One hundred and twelve hospitalised children and their parents (n = 112) were studied. The percentage of positive words used by children to define their nurse was higher in children interviewed after the introduction of non-conventional uniforms (96.2% vs. 81.8%, p = 0.01). Children's perception of nurses was significantly improved by the use of multi coloured attire ('bad'-'good' SDS: p = 0.01; 'disagreeable'-'nice' SDS: p = 0.001). Children's perceptions regarding hospital environment did not change. Parents' perception of nurses' uniform adequacy to the role and capability to reassure resulted improved (p < 0.0001, p = 0.0003). CONCLUSIONS: Multi-coloured non-conventional attire were preferred by hospitalised children and their parents. Their introduction improved the perception children have of their nurses. Moreover, the coloured uniforms improve the parents perception about the reliability of the nurse. RELEVANCE TO CLINICAL PRACTICE: The use of non conventional nurses' attire can contribute to improve the child-nurse relation, which has the potential to ease the discomfort experienced by children due to hospitalisation. PMID- 19284437 TI - Rehabilitation nurses practices in relation to urinary incontinence following stroke: a cross-cultural comparison. AB - AIM: To explore nurses' practices and influences in relation to urinary incontinence following stroke, in the UK, Sweden and China. BACKGROUND: Urinary incontinence following stroke is common, under-recognised and poorly researched. Before appropriate rehabilitation interventions can be developed, an understanding of nurses' current management approaches and cultural influences is required. DESIGN: Qualitative. METHODS: Semi-structured interviews were undertaken with ten registered nurses from at least four different stroke units in three countries (n = 30). Interviews were carried out in the participants' first language, using an agreed interview guide. Following translation, thematic analysis focusing on manifest meaning was undertaken, using an iterative approach involving electronic and face-to-face discussions. RESULTS: The consequence of only superficial assessment was no systematic identification of types or causes of urinary incontinence and no individualised plans developed. A process model of practice, common to all three countries, was identified for stroke survivors with urinary incontinence. Routine core activities were followed by the palliative pathway (most frequently), where urinary incontinence was contained to protect the stroke survivors' safety and ensure social continence; or the rehabilitative route (more rarely), where simple continence promoting activities were implemented with the purpose of facilitating recovery of bladder function. CONCLUSIONS: Nurses' reactively manage urinary incontinence following stroke, adopting a routinised approach based on local custom and practice. Promotion of urinary continence is not a priority area of stroke rehabilitation for nurses in western or eastern countries. RELEVANCE TO CLINICAL PRACTICE: The dearth of evidence-based interventions available to rehabilitate bladder function following stroke means that stroke nursing practice is an experience-based endeavour. This study explains the nurses' focus on containment and social continence and highlights the need to systematically assess stroke survivors' bladder rehabilitation needs, identify types of urinary incontinence and adopt appropriate urinary continence promoting practices. PMID- 19284438 TI - Validity of the symptomatic approach used by nurses in diagnosing vaginal infections. AB - AIMS AND OBJECTIVES: To examine the validity and effectiveness of a symptomatic approach based on symptoms, administered by a nurse working in women's health area, in identifying vaginal infections in women with or without vaginal discharge. DESIGN: Survey. METHODS: Between November 2005-August 2006, 300 married women were included. A questionnaire and vaginal discharge assessment form were used for data collection. The modified algorithm of a symptomatic approach without speculum examination was used to manage participants. Three diagnoses were made for each participant. The first diagnosis was made by the research nurse using the vaginal discharge assessment form; the second diagnosis is the clinical diagnosis which was made after the gynaecological examination by a physician; and the third diagnosis is the microscopic diagnosis made by the physician when assessing the collected specimens. RESULTS: In the methodological part of this study, the sensitivity, specificity, positive predictive value and negative predictive value of the symptomatic approach were 91.5, 69.7, 87.7 and 77.5%, respectively. The symptomatic approach had 27.7% sensitivity for bacterial vaginosis, 16.6% for trichomonas vaginalis and 62.8% for candida albicans, compared to the microscopic results. The sensitivity of the approach was higher for the identification of candidiasis compared to bacterial vaginosis and trichomonas vaginalis. CONCLUSION: From the results, it is suggested that the symptomatic approach model can be used by nurses who work in women's health area to diagnose vaginal infections. RELEVANCE TO CLINICAL PRACTICE: This study showed that trained clinician nurses will be able to diagnose and manage vaginal infections - especially candidiasis - by using the symptomatic approach. Moreover, the symptomatic approach will encourage women not to delay or fail to seek treatment for their health problems, especially for vaginal infections in conservative societies. PMID- 19284439 TI - Commentary on Meyer G, Kopke S, Haasert B & Muhlhauser I (2009) Restraint use among nursing home residents: cross-sectional study and prospective cohort. Journal of Clinical Nursing 18, 981-990 and Saarnio R, Isola A & Laukkala H (2009) The use of physical restraint in institutional care of older people in Finland: nurses' individual, communal and alternative modes of action. Journal of Clinical Nursing 18, 132-140. PMID- 19284442 TI - Autoimmune thyroid diseases: genetic susceptibility of thyroid-specific genes and thyroid autoantigens contributions. AB - Autoimmune thyroid diseases are common polygenic multifactorial disorders with the environment contributing importantly to the emergence of the disease phenotype. Some of the disease manifestations, such as severe thyroid-associated ophthalmopathy, pretibial myxedema and thyroid antigen/antibody immune complex nephritis are unusual to rare. The spectrum of autoimmune thyroid diseases includes: Graves' disease (GD), Hashimoto's thyroiditis (HT), atrophic autoimmune thyroiditis, postpartum thyroiditis, painless thyroiditis unrelated to pregnancy and thyroid-associated ophthalmopathy. This spectrum present contrasts in terms of thyroid function, disease duration and spread to other anatomic location. The genetic basis of autoimmune thyroid disease (AITD) is complex and likely to be due to genes of both large and small effects. In GD the autoimmune process results in the production of thyroid-stimulating antibodies and lead to hyperthyroidism, whereas in HT the end result is destruction of thyroid cells and hypothyroidism. Recent studies in the field of autoimmune thyroid diseases have largely focused on (i) the genes involved in immune response and/or thyroid physiology with could influence susceptibility to disease, (ii) the delineation of B-cell autoepitopes recognized by the main autoantigens, thyroglobulin, thyroperoxidase and TSH receptor, to improve our understanding of how these molecules are seen by the immune system and (iii) the regulatory network controlling the synthesis of thyroid hormones and its dysfunction in AITD. The aim of the present review is to summarize the current knowledge regarding the relation existing between some susceptibility genes, autoantigens and dysfunction of thyroid function during AITD. PMID- 19284443 TI - Comparison of IL10 and IL2 genotypes of Turkish population with other populations. AB - The human genome has been shaped by evolutionary and historical forces. Therefore, genetic polymorphisms are useful tools not only to understand the susceptibility to disease in modern populations, but the history of ancestral populations as well. For this purpose, data on genetic polymorphisms such as human leucocyte antigen, mitochondrial DNA sequence variability and the frequencies of TAP1 and TAP2 gene variants in Turkey have been reported previously. Here we have used interleukin (IL)-10 (-592C/A, -819T/C, -1082G/A) and IL-2 (-330T/G) as genetic markers to study the relationship between Turkish population and other populations. PMID- 19284444 TI - Cytotoxic T lymphocyte-associated molecule-4 polymorphism in Turkish patients with Hashimoto thyroiditis. AB - We previously shown that in a Turkish population, the A/G polymorphism in exon 1 of the cytotoxic T cell lymphocyte-associated molecule-4 (CTLA-4) gene is associated with Graves' disease, and that the G allele may contribute to susceptibility for developing Graves' disease. This polymorphism was identified in 197 patients with Hashimoto thyroiditis (HT) (126 women, 71 men; aged, 42.92 +/- 13.4 years) and 98 healthy individuals (56 women, 21 men; aged, 42.27 +/- 13.43 years) in Turkish population. Polymorphisms were analysed using a polymerase chain reaction-restriction fragment length polymorphism method. Frequency of the A/G genotypes was not significantly different in patients with HT when compared with controls in both sexes (P > 0.05). There was no statistical difference in age, sex, cigarette smoking, initial serum thyroid hormone levels, initial goiter size and thyroid autoantibodies among the patients with the three different genotypes (G/G, A/G and A/A). We concluded that A/G polymorphism of CTLA molecule is linked to occurrence of Graves' disease bu not to HT in the Turkish population. PMID- 19284445 TI - The association of CTLA-4 and CD28 gene polymorphisms with idiopathic ischemic stroke in the paediatric population. AB - Autoimmune vasculitis is believed to be a critical factor in the development of idiopathic childhood ischemic stroke. The association of polymorphisms in CTLA-4 and CD28 with some immune vasculitides, such as systemic lupus erythematosus (SLE) and Behcet's disease has been reported. The aim of the present study is to investigate the association of the genetic variants in the CTLA-4 and CD28 genes of children who suffered idiopathic ischemic stroke using a case-control design. Two single nucleotide polymorphisms (SNPs) in the CTLA-4 gene and an SNP in the CD28 gene were genotyped in 51 patients who suffered idiopathic ischemic stroke, and in 74 healthy controls from mainland China. An SNP, CTLA-4+49A/G located in exon 1 of the CTLA-4 gene, showed nominal association with the disease (P = 0.012, odds ratio (OR) = 2.09, 95% confidence interval (CI) = 1.17-3.73) using allele-based analysis. Homozygous carriers of the G allele of this SNP were more common in the patients than in the controls (P = 0.008). The CD28IVS3 +17TT genotype was found to be more common in the patients than in the controls (P = 0.039, OR = 2.96, 95% CI = 1.02-8.58). No correlations of at-risk genotype (G/G) of CTLA-4+49A/G and genotype (T/T) of CD28+17T/C with the main clinical features of idiopathic childhood ischemic stroke were observed. The results suggest that polymorphisms in the CTLA-4 and CD28 genes may contribute to the increased risk of idiopathic ischemic stroke. PMID- 19284446 TI - New allele name of some HLA-DRB1*1401: HLA-DRB1*1454. AB - The primary function of MHC polymorphism is considered as the foundation of self defense mechanism of the host in surveillance against countless diverse invading pathogens. However, this biological function can also elicit undesirable immunological responses that jeopardize transplantations when compatibility between donors and recipients is unfavourable. PMID- 19284447 TI - Identification of a novel HLA-A allele, A*1131, in a Taiwanese. AB - Here we report the identification and sequence analysis of a new HLA-A11* variant, A*1131 allele, found in a Taiwanese volunteer bone marrow donor. The novel A*11 variant is identical to A*1125 in exon 2 but differs from A*1125 in exon 3 by one nucleotide substitution at position 527 causing an amino acid change at codon 152 E-->V (GAG-->GTG). In comparison with HLA-A*110101, allele A*1131 has three nucleotide differences in exon 3: 527 C-->T, 538 C-->T and 539 A ->T leading to two amino acid variations at residues 152 A-->V and 156 Q-->L. PMID- 19284449 TI - Are there predominant strains and toxins of Staphylococcus aureus in atopic dermatitis patients? Genotypic characterization and toxin determination of S. aureus isolated in adolescent and adult patients with atopic dermatitis. AB - The colonization of Staphylococcus aureus is one of the most important aggravating factors of atopic dermatitis (AD). Until now, the importance of S. aureus in AD and a positive correlation between colonization with S. aureus and clinical severity/skin barrier function has been demonstrated. The aim of this study was to determine whether there are certain clones of S. aureus which colonize the skin of AD patients. For this purpose, the genotype of S. aureus isolated from AD patients was examined by newly-developed typing methods. With 36 strains of S. aureus isolated from 35 patients with AD, spa typing, multi-locus sequence typing (MLST), and staphylococcal toxin gene assay by multiplex polymerase chain reaction, were performed. Clinical severity and skin barrier function were evaluated with eczema area and severity index (EASI) and with transepidermal water loss (TEWL). Among 36 strains of S. aureus, 14 sequence types (ST) and 20 spa types were identified, suggesting a very heterogeneous genetic composition of S. aureus and the absence of a prevailing genotype in S. aureus colonized with AD patients. Furthermore, there was no specific genotype of S. aureus which was associated with the clinical severity of AD or skin barrier dysfunction. A toxin gene assay, however, showed the predominance of S. aureus strains carrying sea and/or tsst-1. To the best of our knowledge, this is the first report to show the genetic composition of S. aureus strains isolated from AD patients determined by sequence-based typing methods. PMID- 19284450 TI - Scalp dermoscopy of androgenetic alopecia in Asian people. AB - Although dermoscopy is used mainly for diagnosing pigmented skin lesions, this device has been reported to be useful in observing alopecia areata and frontal fibrosing alopecia. Herein, we investigated the dermoscopic features and their incidence of androgenetic alopecia (AGA; n = 50 men) and female AGA (FAGA; n = 10 women) in Asian people. More than 20% hair diameter diversity (HDD), which reportedly is an early sign of AGA and corresponds to hair follicle miniaturization, was observed in the affected area of all AGA and FAGA cases, suggesting that HDD is an essential feature to diagnose AGA and FAGA. Peripilar signs, corresponding to perifollicular pigmentation, were seen in 66% (33/50) of AGA and 20% (2/10) of FAGA women. This incidence in the present study was lower than previously reported in white subjects possibly because the Asian skin color conceals slight peripilar pigmentation. Yellow dots were observed in 26% (13/50) of AGA and 10% (1/10) of FAGA cases and the number of yellow dots in AGA and FAGA was limited to 10 on the overall hair loss area. Yellow dots possibly indicate the coincidence of AGA and enlargement of the sebaceous glands caused by common end-organ hypersensitivity to androgen. In conclusion, dermoscopy is useful to diagnose AGA and FAGA and provides insights into the pathogenesis of AGA. PMID- 19284451 TI - Case of creeping disease treated with ivermectin. AB - We report a case of creeping disease treated successfully with ivermectin. A 46 year-old man presented with a 1-month history of pruriginous linear erythema on his right thigh after a visit to Indonesia. Although he had no history of eating raw fish or meat, he walked along the river and in the jungle without wearing shoes. Creeping disease caused by animal hookworm was strongly suspected. The presence of parasite larvae was not confirmed in biopsied skin specimens. In enzyme-linked immunosorbent assay, serum samples were negative for binding to hookworm antigens, including Ancylostoma canium, Necator americanus and Gnathostoma doloresi. He was treated with a single 12 mg oral dose (200 microg/kg) of ivermectin. The eruption and pruritus resolved within a few days after the administration and did not relapse. PMID- 19284452 TI - Fluorescence navigation with indocyanine green for detecting sentinel nodes in extramammary Paget's disease and squamous cell carcinoma. AB - The radioisotope navigation method, which has usually been used for identification of sentinel nodes, is less useful in locating sentinel nodes close to primary lesions in cases of extramammary Paget's disease because of overlapping radioactivity from the primary site. We applied fluorescence navigation with indocyanine green (ICG) in two patients with skin cancer to cover this defect. The use of a charge-coupled device camera enabled real-time visualization of dynamic lymph flow without skin incision. The inguinal skin over the identified sentinel node with a handheld gamma probe was in accordance with the point detected by ICG fluorescence in a patient with squamous cell carcinoma of the foot. Sentinel node biopsy using fluorescence navigation with ICG proved to be easy and reliable. PMID- 19284453 TI - Case of herpes zoster duplex bilateralis. AB - Non-contiguously simultaneous development of herpes zoster is very rare. It is named either herpes zoster duplex unilateralis or bilaterarlis, depending on whether one or both sides of the body are involved. Herein, we report a 21-year old man, who had been treated for ulcerative colitis with prednisolone, and presented with painful grouped vesicles of the lower abdomen and back in a relatively symmetrical distribution. A Tzanck smear and punch biopsy were performed on the vesicles of the back. We report a rare case of symmetrical herpes zoster duplex bilateralis. PMID- 19284454 TI - Case of cutaneous Scedosporium apiospermum infection successfully treated with voriconazole. AB - We report a case of cutaneous infection due to Scedosporium apiospermum in a 75 year-old immunocompromised male patient who had received long-term corticosteroid and immunosuppressant therapy for the treatment of nephrotic syndrome. The patient came to our department complaining of erythema with a number of pustules on the dorsal surface of the right hand. S. apiospermum was identified from a culture taken from the pus. After unsuccessful treatment with topical ketoconazole, oral itraconazole and oral terbinafine, the lesion quickly resolved with the daily administration of 400 mg voriconazole. No recurrence was observed despite discontinuation of voriconazole due to drug-induced hepatitis. Voriconazole holds out the promise of an effective treatment for invasive Scedosporium infection. PMID- 19284455 TI - Case of lupus vulgaris diagnosed 50 years after onset. AB - Cutaneous tuberculosis is an infrequent form of extrapulmonary tuberculosis, but is a symptom that can lead to diagnosis of tuberculosis. We describe a case of lupus vulgaris in a 79-year-old woman who had a 50-year history of a slowly growing plaque on her right cheek. She visited many hospitals without resolution and the plaque gradually enlarged. Recently, she was misdiagnosed with eczema and prescribed topical steroids that had no effect, and she subsequently visited our outpatient clinic. A diagnosis of lupus vulgaris was made based on histopathology, culture and polymerase chain reaction, and isoniazid, rifampicin and ethambutol were administered as antituberculosis treatment. Although the incidence of cutaneous tuberculosis has decreased significantly in developed countries, knowledge and awareness of the disease are still of importance for proper diagnosis and treatment. PMID- 19284456 TI - Skin metastasis of prostate adenocarcinoma to glans penis showing no correlation with serum prostate-specific antigen level. PMID- 19284457 TI - Case of acquired trichorrhexis nodosa: scanning electron microscopic observation. PMID- 19284458 TI - Prominent facial edema caused by potassium iodide solution in a patient with sporotrichosis. PMID- 19284459 TI - Successful treatment of multiple eccrine hidrocystoma with topical atropine sulfate ointment. PMID- 19284460 TI - Lichen amyloidosis of the auricular concha: successful treatment with electrodesiccation. PMID- 19284461 TI - Acne conglobata successfully treated by fractional laser after CO laser abrasion of cysts combined with topical tretinoin. PMID- 19284462 TI - Glycylglycine decreases the size of conspicuous facial pores: Single-blinded half areas of face-applied study. PMID- 19284464 TI - Better birthing, better babies. Abstracts of the 13th Annual Congress of the Perinatal Society of Australia and New Zealand. April 19-22, 2009. Darwin, Northern Territory, Australia. PMID- 19284466 TI - Will history repeat itself? PMID- 19284467 TI - Sexual function: diagnosis and management of sexual dysfunction. PMID- 19284468 TI - The use of estrogen therapy in women's sexual functioning (CME). AB - INTRODUCTION: Estrogen is relevant to women's well-being including sexual functioning. Aim. The goal of this Continuing Medical Education article was to provide a comprehensive review of the effect of exogenous estrogen use on women's sexual function. Main Outcome Measures. We present a literature review. METHODS: The medical literature was accurately searched (1990-2008) with regard to estrogen therapy in menopausal women by using several terms related to and including the terms "estrogen" and "sexual function." RESULTS: A review of the studies most useful to guide menopausal practice. CONCLUSIONS: Estrogen decline is one of the key factors contributing to sexual functioning during menopausal transition and beyond. Systemic estrogen treatments are associated with significant benefits in some domains of menopausal sexual function, especially when estradiol is delivered transdermally, whereas local estrogens are effective in preventing urogenital aging. Even tibolone, a selective tissue estrogenic activity regulator, displays positive effects in postmenopausal women with sexual complaints. However, a tailored approach to the individual woman is always needed. PMID- 19284470 TI - The Henry mummy wrap and the Henry finger sweep surgical techniques. PMID- 19284472 TI - Correlation of degree of penile curvature between patient estimates and objective measures among men with Peyronie's disease. AB - INTRODUCTION: : Among men with Peyronie's disease (PD), the degree of penile curvature has significant implications on psychological well-being, sexual function, treatment planning, and posttherapy evaluations. AIM: The primary objective of the current study was to correlate patients' estimates of penile angulation with objective measures. MAIN OUTCOME MEASURES: (i) Proportion of patients over- or underestimating their actual degree of curvature; and (ii) degree differences between patient estimates and objective measures of penile curvature. Methods. At baseline, patients with established PD were asked to provide a "best estimate" of their degree of penile curvature. Objective measures of penile angulation were then performed using standardized photographs and protractor-based measurement of penile curvature during full erection. Correlations were performed between patient estimates of penile curvature and objective measures of penile angulation. RESULTS: Eighty-one men with established PD and a mean age of 52 years (range: 20-72 years) were prospectively evaluated. Mean duration of disease was 33 months (range: 6-276 months), and mean plaque size was 1.4 cm +/- 0.1 standardized error (SE). The proportion of patients with dorsal, lateral, and ventral curvatures was 39%, 57%, and 4%, respectively. Patient estimates of baseline penile curvature (mean 51 degrees +/- 3.1 SE) differed significantly from objective measurements (mean 40 degrees +/- 2.4, P = 0.001). A significantly higher proportion of patients overestimate their actual degree of penile curvature (54% overestimate, 26% underestimate, and 20% are accurate within 5 degrees, P = 0.002). Compared with objective measures, patients' estimates of degree of penile curvature differed by an average of 20 degrees +/- 2.2 SE. CONCLUSIONS: Patients with PD tend to overestimate their degree of penile curvature. Objective measurement of penile angulation is necessary to accurately counsel patients regarding disease severity, recommend appropriate treatment strategies, and objectively evaluate outcomes following therapy. PMID- 19284473 TI - The influence of environment and life-history traits on the distribution of genes and individuals: a comparative study of 11 rainforest trees. AB - This study investigates patterns of genetic connectivity among 11 co-distributed tropical rainforest tree species from the genus Elaeocarpus across a biogeographic barrier, the Black Mountain Corridor (BMC) in the Australian Wet Tropics (AWT). We analysed a combination of allelic and flanking region sequence data from microsatellite markers, and evaluated the relative influence of environmental preferences and functional traits on genetic diversity and gene flow. The results indicate that only in three species geographic structuring of haplotype distribution reflects a north vs. south of the BMC pattern. Environmental factors linked with altitude were recognized as affecting genetic trends, but the selective processes operating on upland species appear to be associated with competitiveness and regeneration opportunities on poor soil types rather than climate variables alone. In contrast to previous observations within southeastern Australian rainforests, genetic differentiation in the AWT appears to be associated with small-fruited rather than large-fruited species, highlighting how external factors can influence the dispersal dimension. Overall, this study emphasizes the importance of considering functional and environmental factors when attempting generalizations on landscape-level patterns of genetic variation. Understanding how plant functional groups respond to environmental and climatic heterogeneity can help us predict responses to future change. PMID- 19284474 TI - More introgression with less gene flow: chloroplast vs. mitochondrial DNA in the Picea asperata complex in China, and comparison with other Conifers. AB - Recent work has suggested that rates of introgression should be inversely related to levels of gene flow because introgressed populations cannot be 'rescued' by intraspecific gene flow if it is too low. Mitochondrial and chloroplast DNA (mtDNA and cpDNA) experience very different levels of gene flow in conifers due to their contrasted maternal and paternal modes of transmission, hence the prediction that mtDNA should introgress more readily than cpDNA in this group. Here, we use sequence data from both mtDNA and cpDNA to test this hypothesis in a group of closely related spruces species, the Picea asperata complex from China. Nine mitochondrial and nine chloroplast haplotypes were recovered from 459 individuals in 46 natural populations belonging to five species of the Picea asperata complex. Low variation was found in the two mtDNA introns along with a high level of differentiation among populations (G(ST) = 0.90). In contrast, we detected higher variation and lower differentiation among populations at cpDNA markers (G(ST) = 0.56), a trend shared by most conifer species studied so far. We found that cpDNA variation, although far from being fully diagnostic, is more species-specific than mtDNA variation: four groups of populations were identified using cpDNA markers, all of them related to species or groups of species, whereas for mtDNA, geographical variation prevails over species differentiation. The literature suggests that mtDNA haplotypes are often shared among related conifer species, whereas cpDNA haplotypes are more species-specific. Hence, increased intraspecific gene flow appears to decrease differentiation within species but not among species. PMID- 19284475 TI - Regulation and function of neuronal GTP-Ras in facial motor nerve regeneration. AB - Activation of Ras into the GTP-binding, 'ON' state is a key switch in the neurotrophin-mediated neuronal survival and neurite outgrowth, in vitro as well as in vivo. In the current study we explored changes in GTP-Ras levels following facial nerve injury and the ensuing regeneration and the effects of perturbing these changes in vivo using synapsin-promoter mediated neuronal expression of constitutively active Val12H-Ras (synRas). Quantification of GTP-Ras and total Ras revealed a precipitous drop in the relative GTP-Ras levels in the axotomized facial motor nucleus, to 40% of normal levels at 2 days after cut, followed by a partial recovery to 50-65% at 4-28 days. On western blots, control and axotomized nuclei from synRas mutants showed a 2.2- and 2.5-fold elevation in GTP-Ras, respectively, compared with their wild type littermate controls (p < 5%, anova, TUKEY post-hoc), with the levels in the axotomized synRas nucleus slightly but not significantly above that in the uninjured littermate control (p = 9.9%). Similar increase was also observed in the pERK but not pAKT targets of the Ras cascade. This moderate elevation of GTP-Ras strongly curtailed post-traumatic neuronal cell death (-65%), the influx of T-cells (-48%) as well as other parameters of neuroinflammatory response. Although synRas did not affect the speed of axonal regeneration or functional recovery it caused a very pronounced increase in central axonal sprouting. These current data emphasize the role of reduced active Ras, and by extension, the reduced overall level of retrograde neurotrophin signalling after axotomy, in mediating post-traumatic cell death and inflammation and in restricting the sprouting response. Moreover, the neuroprotective and central sprouting-enhancing effects of neuronal Val12H-Ras could help promote recovery in CNS injury. PMID- 19284476 TI - The expression of P-glycoprotein is increased in vessels with blood-brain barrier impairment in a stroke-prone hypertensive model. AB - AIMS: We previously reported that the blood-brain barrier (BBB) function was impaired in vessels in the hippocampus in 3-month-old stroke-prone spontaneously hypertensive rats (SHRSP). In this study, we examined gene and protein expressions of P-glycoprotein, a representative efflux transporter of cerebral vessels, in the BBB-damaged hippocampal vessels of SHRSP and in the vessels of Wistar Kyoto (WKY) rats as controls, to clarify roles of the efflux transporter in the BBB-damaged vessels. METHODS: The expression of P-glycoprotein in hippocampal and cortical samples was examined by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunoelectron microscopic techniques. RESULTS: Real-time RT-PCR and Western blotting analyses revealed that the gene and protein expressions of P glycoprotein were increased in the hippocampal samples of 3-month-old SHRSP compared with hippocampal samples of 3-month-old WKY rats or with cortical samples of SHRSP. The gene expression of P-glycoprotein was also increased in the hippocampal samples of 4-week-old SHRSP. Immunoelectron microscopic examination revealed that immunosignals of P-glycoprotein were seen in the luminal and ab luminal cytoplasmic membranes of endothelial cells and the basal lamina, that the labelling density of P-glycoprotein in the vessel wall was higher in the hippocampus of 3-month-old SHRSP than in other groups and that the immunosignals of P-glycoprotein were occasionally co-located with those of albumin. CONCLUSIONS: These findings indicate that the expression of P-glycoprotein is increased in BBB-damaged hippocampal vessels in hypertensive SHRSP compared with those in WKY rats. PMID- 19284477 TI - Mineralocorticoid receptor mRNA expression is increased in human hippocampus following brief cerebral ischaemia. AB - BACKGROUND: We have previously reported that neuronal endangerment in vitro and hypothermic transient global ischaemia in vivo each result in increased mineralocorticoid receptor (MR) expression. In both models MR induction is associated with increased neuronal survival, and blocking MR signalling reduces neuronal survival. Furthermore, transgenic overexpression of human MR promotes neuronal survival both in vitro and in vivo. AIMS: Here we have assessed whether brief periods of cerebral ischaemia in human subjects, such as occurs in cardiac arrest from which successful resuscitation is achieved, are associated with a sustained increase in hippocampal MR mRNA expression. METHODS: Human post-mortem brain sections from patients who had died in the weeks following cardiac arrest were analysed for MR mRNA expression by in situ hybridization. RESULTS: Sustained upregulation of MR mRNA expression was observed in the dentate gyrus region of human hippocampus following a brief episode of cerebral ischaemia. CONCLUSIONS: This confirms that MR mRNA expression is regulated following neuronal injury in human brain, and suggests that the benefits of increased MR expression seen in animal models of ischaemia may also be observed in humans. PMID- 19284478 TI - Microcalcification after excitotoxicity is enhanced in transgenic mice expressing parvalbumin in all neurones, may commence in neuronal mitochondria and undergoes structural modifications over time. AB - AIMS: Parenchymal microcalcification in the brain coincides with neurodegenerative diseases, but is also frequently found in neurologically normal individuals. The origin and role of this process are still under debate. Parvalbumin (PV) is a protein acting as a Ca(2+) buffer and Ca(2+) shuttle towards intracellular Ca(2+) sinks, like mitochondria and the endoplasmic reticulum. Constitutively, it is present in a subset of inhibitory neurones. In transgenic mice expressing pan-neuronal PV, the mitochondrial volume is reduced. We tested whether elevated levels of intraneuronal [Ca(2+)] and reduced mitochondrial volume in the neurone interfere with the generation of parenchymal microcalcification. METHODS: The striatum of wild type and transgenic mice was injected with the glutamate receptor agonist ibotenic acid (IBO), which is known to induce not only excitotoxic neurodegeneration, but also parenchymal calcification. Sections were studied by light and electron microscopy at various time points after IBO application. RESULTS: Morphometric analysis 2, 4 and 20 weeks after IBO application revealed microcalcification in transgenic and wild type mice; the calcification process, however, was enhanced and accelerated in the transgenic animals. Ultrastructural analyses suggest neuronal mitochondria as the nucleators of the deposits which consist of hydroxyapatite. The time dependent changes in size and surface structure of the deposits indicate the presence of biological mechanisms in the brain promoting regression of bioapatites. CONCLUSIONS: The overload of intraneuronal [Ca(2+)] in combination with impaired mitochondrial function activates neuronal microcalcification. It is hypothesized that this process is an alternative/adaptive mechanism of the neurone to reduce further brain damage. PMID- 19284479 TI - Expression of reticulon 3 in Alzheimer's disease brain. AB - AIMS: Reticulon 3 (RTN3), a member of the reticulon family of proteins, interacts with the beta-secretase, beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1), and inhibits its activity to produce beta-amyloid protein. The aim of the present study was to clarify the biological role of RTN3 in the brain and its potential involvement in the neuropathology of Alzheimer's disease (AD). METHODS: We performed immunohistochemical and biochemical analyses using a specific antibody against RTN3 to investigate the expression and subcellular localization of RTN3 in control and AD brain tissue samples. RESULTS: Western blot analysis revealed no significant differences in the RTN3 levels between control and AD brains. Immunohistochemical staining showed that RTN3 immunoreactivity was predominantly localized in pyramidal neurones of the cerebral cortex. The patterns of RTN3 immunostaining were similar in control and AD cerebral cortices, and senile plaques were generally negative for RTN3. Biochemical subcellular fractionation disclosed that RTN3 colocalized with BACE1 in various fractions, including the endoplasmic reticulum and the Golgi apparatus. Double immunofluorescence staining additionally indicated that RTN3 was localized in both endoplasmic reticulum and Golgi compartments in neurones. CONCLUSIONS: These results show that RTN3 is primarily expressed in pyramidal neurones of the human cerebral cortex and that no clear difference of RTN3 immunoreactivity is observable between control and AD brains. Our data also suggest that there is considerable colocalization of RTN3 with BACE1 at a subcellular level. PMID- 19284480 TI - Altered arginine metabolism in the central nervous system (CNS) of the Cln3-/- mouse model of juvenile Batten disease. AB - BACKGROUND: Juvenile neuronal ceroid lipofuscinoses (JNCL) or juvenile Batten disease is a recessively inherited childhood neurodegenerative disorder resulting from a mutation in CLN3, which encodes a putative lysosomal protein of unknown function. AIM: Recent evidence suggests that a disruption in CLN3 function results in altered regulation of arginine transport into lysosomes, and may influence intracellular arginine levels. We sought to investigate the possible consequences of arginine dysregulation in the brain of the Cln3(-/-) mouse model of JNCL. METHODS: Using a combination of enzyme assays, metabolite profiling, quantitative reverse-transcription polymerase chain reaction and Western blotting, we analysed the activities and expression of enzymes involved in arginine metabolism in the cerebral cortex and cerebellum of Cln3(-/-) mice over several developmental time points. RESULTS: We report subtle, but significant changes in the activities of enzymes involved in the citrulline-NO recycling pathway, and altered regulation of neuronal nitric oxide synthase in the cortex and cerebellum of Cln3(-/-) mice. In addition, a significant decrease in arginine transport into cerebellar granule cells was observed, despite an apparent upregulation of the cationic amino acid transporter-1 transporter at the cell surface. Our results provide further evidence that CLN3 function and arginine homeostasis are intricately related, and that cellular mechanisms may act to compensate for the loss of this protein. CONCLUSIONS: This and other studies indicate that CLN3 dysfunction in JNCL may result in multiple disturbances in metabolism that together contribute to the pathophysiological processes underlying this disease. PMID- 19284481 TI - Epidermal growth factor receptor extracellular domain mutations in primary glioblastoma. AB - AIMS: Novel missense mutations of the epidermal growth factor receptor (EGFR) extracellular domain have been recently described in a large series of glioblastomas. METHODS: The exons 2, 3, 7, 8 and 15 coding for the EGFR extracellular domain were sequenced in a series of 161 consecutive primary glioblastomas and correlated with clinical features of patients in order to determine whether these alterations are linked to specific clinical characteristics of the disease. RESULTS: Missense mutations were observed in 18 cases (11.2%), and 4 novel mutations were detected, including G178C, A271C, C818A and C1860G. Mutations of the EGFR extracellular domain were not associated with overall survival or with age at onset of the disease. In contrast, the EGFR extracellular domain mutations were significantly associated with patients' gender. Indeed, 15 mutations were observed in men vs. 3 in women (P < 0.05). EGFR extracellular domain mutations were also strongly associated with EGFR amplification (P < 0.0001). CONCLUSIONS: To our knowledge, EGFR extracellular domain mutations are the first genomic abnormalities associated with gender in primary glioblastomas, although a link between mutations of the EGFR tyrosine kinase domain and gender has been previously made in lung cancer. PMID- 19284482 TI - Transformation of a low-grade pineal parenchymal tumour to secondary pineoblastoma. PMID- 19284483 TI - An unusual cause of dysphagia and dysarthria. PMID- 19284485 TI - Methadone for chronic pain in older adults: blast from the past but are we ready for it to return to prime time? PMID- 19284486 TI - Psychological factors and domains of neck pain disability. AB - OBJECTIVE: This study evaluated the relationship between psychological distress and disability associated with neck pain, analyzed the Neck Disability Index (NDI) for disability factors, and assessed the impact of psychological distress on those domains of disability. DESIGN: Prospective cross-sectional analytic survey. SETTING: Outpatient physical therapy clinic. PATIENTS: Sixty-one consecutive adult subjects with dominant neck pain participated. OUTCOME MEASURES: Each subject completed the NDI, psychometric measures for the Distress Risk Assessment Method, and a numeric pain rating scale. RESULTS: Measures of depression, somatization, and pain intensity explained 60% of the variance of disability due to neck pain. Factor analysis revealed two disability factors in the NDI dealing with physical activity/participation limitations and nonphysical activity-related impairments in bodily function. Psychological distress and pain intensity explained 25.6% of the variance of the factor dealing with activity/participation limitations, and 53.5% of the variance for the factor associated with impairments in bodily functions. CONCLUSION: Nondistressed adults reported significantly less disability due to neck pain than psychologically distressed subjects. The NDI was found to contain two factors that pertain to three domains of the disability. Five items relating to impairments in bodily function strongly correlated with depression and somatization. Presence of psychological distress has a confounding effect on NDI scores. An outcome measure containing items related only to activity limitations and participation restrictions might give a truer picture of disability associated with neck pain for patients with psychological distress. PMID- 19284487 TI - Are spirituality and religiosity resources for patients with chronic pain conditions? AB - OBJECTIVE: We studied whether or not spirituality/religiosity is a relevant resource for patients with chronic pain conditions, and to analyze interrelations between spirituality/religiosity (SpREUK Questionnaire; SpREUK is an acronym of the German translation of "Spiritual and Religious Attitudes in Dealing with Illness"), adaptive coping styles that refer to the concept of locus of disease control (AKU Questionnaire; AKU is an acronym of the German translation of "Adaptive Coping with Disease"), life satisfaction, and appraisal dimensions. PATIENTS: In a multicenter cross-sectional study, 580 patients with chronic pain conditions were enrolled. RESULTS: We found that the patients relied on both external powerful sources of disease control and on internal powers and virtues, while Trust in Higher Source (intrinsic religiosity) or Illness as Chance (reappraisal) were valued moderately; Search for Meaningful Support/Access (spiritual quest orientation) was of minor relevance. Stepwise regression analyses revealed that the internal sources of disease control, such as Conscious and Healthy Way of Living and Positive Attitudes, were (apart from the religious denomination) the strongest predictors of patients' reliance on spirituality/religiosity. Both behavioral styles were rated significantly lower in patients who regarded themselves as neither religious nor spiritual. Positive disease interpretations such as Challenge and Value were clearly associated with a spiritual quest orientation and intrinsic religiosity. CONCLUSION: The associations between spirituality/religiosity, positive appraisals. and internal adaptive coping strategies indicate that the utilization of spirituality/religiosity goes far beyond fatalistic acceptance, but can be regarded as an active coping process. The findings support the need for further research concerning the contributions of spiritual coping in adjustment to chronic pain. PMID- 19284488 TI - Efficacy of 5-day continuous lidocaine infusion for the treatment of refractory complex regional pain syndrome. AB - OBJECTIVE: Chronic regional pain syndrome (CRPS) is a severe pain condition that usually results from an injury or surgical procedure. The pain in CRPS often spreads from the site of injury, and with time becomes refractory to conventional therapy. The present study was undertaken to evaluate the effects of 5-day continuous intravenous lidocaine treatment in patients afflicted with CRPS. METHODS: Intravenous lidocaine was administered in an escalating dose schedule to 49 severely affected CRPS patients in a monitored setting over 5 days. Evaluation of pain parameters and other signs and symptoms of CRPS were obtained during the infusion and at 1, 3, and 6 months following therapy. RESULTS: The majority of patients demonstrated a significant decrease in pain parameters and other symptoms and signs of CRPS. The pain reduction lasted an average of 3 months. Lidocaine may be particularly effective for thermal and mechanical allodynia. Less clinically significant effects were documented on the motor aspects of the syndrome. DISCUSSION: Intravenous lidocaine administration titrated to 5 mg/L demonstrated: 1) a significant decrease in mechanical and thermal allodynia for three months, 2) lessened associated inflammatory components of CRPS, and 3) only minimal side effects and no severe complications. PMID- 19284489 TI - Betamethasone: friend (soluble), foe (particulate), or either? PMID- 19284492 TI - Immune and nervous systems share molecular and functional similarities: memory storage mechanism. AB - One of the most complex and important features of both the nervous and immune systems is their data storage and retrieval capability. Both systems encounter a common and complex challenge on how to overcome the cumbersome task of data management. Because each neuron makes many synapses with other neurons, they are capable of receiving data from thousands of synaptic connections. The immune system B and T cells have to deal with a similar level of complexity because of their unlimited task of recognizing foreign antigens. As for the complexity of memory storage, it has been proposed that both systems may share a common set of molecular mechanisms. Here, we review the molecular bases of memory storage in neurons and immune cells based on recent studies and findings. The expression of certain molecules and mechanisms shared between the two systems, including cytokine networks, and cell surface receptors, are reviewed. Intracellular signaling similarities and certain mechanisms such as diversity, memory storage, and their related molecular properties are briefly discussed. Moreover, two similar genetic mechanisms used by both systems is discussed, putting forward the idea that DNA recombination may be an underlying mechanism involved in CNS memory storage. PMID- 19284493 TI - A reply to Dembic: on an end to the beginning of misunderstanding the immune response. AB - We all agree that dealing with the complexity and volume of the data necessitates the use of computer modelling. This in turn requires a heuristic conceptual framework to guide this modelling. The first attempt to do this by Cohn has been criticized by Dembic as being severely lacking. This commentary deals with his criticism of the framework to show why Cohn's postulates, in fact, remain unchallenged. PMID- 19284494 TI - Placing regulatory T cells into global theories of immunity: an analysis of Cohn's challenge to integrity (Dembic). AB - In broadening the integrity model, Zlatko Dembic provided one of the few plausible explanations for the existence of regulatory T cells that has been postulated to date and at the same time highlighted deficiencies of the associative antigen recognition model. In defending the virtues of associative antigen recognition, Melvin Cohn has challenged the integrity model and the concept that regulatory T cells have a role in defining the specificity of immune responses. The critique of Cohn's analysis I present here suggests that a greater consideration of quantitative evolutionary constraints removes most of the challenges to integrity. PMID- 19284495 TI - Susceptibility to Yersinia pseudotuberculosis infection is linked to the pattern of macrophage activation. AB - T helper 1 cells play a crucial role in the clearance of Yersinia pseudotuberculosis infection. By producing cytokines and presenting antigens to T cells, activated macrophages can orientate the adaptive immune response. The pathway used by macrophages to metabolize arginine has been employed as an important parameter to discriminate their activation state. In this study, the pattern of macrophage activation in Y. pseudotuberculosis-infected BALB/c (Yersinia-susceptible) and C57BL/6 (Yersinia-resistant) mice and their immunostimulatory capacity were analysed. In the early phase of infection, macrophages obtained from C57BL/6 mice produced higher levels of NO, lower arginase activity, and larger amounts of IL-12 and TNF-alpha than macrophages from BALB/c mice. On the other hand, macrophages derived from BALB/c mice produced higher levels of IL-10 and TGF-beta than C57BL/6 mice. The Y. pseudotuberculosis infection leads to a fall in the macrophage immunostimulatory capacity of both strains of mice, with T-cell proliferation significantly reduced 12 h after infection. Moreover, we observed in the supernatant of co-culture of macrophages from infected mice with T lymphocytes from heat-killed Yersinia immunized mice lower IFN-gamma production by cells from BALB/c mice than by C57BL/6 mice, and IL-4 was produced only by BALB/c mice on the first- and third day post-infection. These results suggest that the pattern of macrophage activation is associated with susceptibility and resistance to Y. pseudotuberculosis infection in BALB/c and C57BL/6 mice. PMID- 19284496 TI - Dendritic cells engineered to express defined allo-HLA peptide complexes induce antigen-specific cytotoxic T cells efficiently killing tumour cells. AB - Most tumour-associated antigens (TAA) are non-mutated self-antigens. The peripheral T cell repertoire is devoid of high-avidity TAA-specific cytotoxic T lymphocytes (CTL) due to self-tolerance. As tolerance is major histocompatibility complex-restricted, T cells may be immunized against TAA presented by a non-self human leucocyte antigen (HLA) molecule and transferred to cancer patients expressing that HLA molecule. Obtaining allo-restricted CTL of high-avidity and low cross-reactivity has, however, proven difficult. Here, we show that dendritic cells transfected with mRNA encoding HLA-A*0201, efficiently present externally loaded peptides from the antigen, Melan-A/MART-1 to T cells from HLA-A*0201 negative donors. CD8(+) T cells binding HLA-A*0201/MART-1 pentamers were detected already after 12 days of co-culture in 11/11 donors. The majority of cells from pentamer(+) cell lines were CTL and efficiently killed HLA-A*0201(+) melanoma cells, whilst sparing HLA-A*0201(+) B-cells. Allo-restricted CTL specific for peptides from the leukaemia-associated antigens CD33 and CD19 were obtained with comparable efficiency. Collectively, the results show that dendritic cells engineered to express defined allo-HLA peptide complexes are highly efficient in generating CTL specifically reacting with tumour-associated antigens. PMID- 19284497 TI - Induction of TLR tolerance in human macrophages by adiponectin: does LPS play a role? AB - Obesity is regarded as a pro-inflammatory state. It is associated with low circulating levels of the adipokine, adiponectin, which is considered to be an anti-inflammatory. However, adiponectin knockout mice do not consistently demonstrate pro-inflammatory phenotypes, suggesting more complexity in the in vivo immunomodulatory effects of adiponectin than originally anticipated. Moreover, adiponectin exerts pro-inflammatory effects in some experimental systems. This contradiction has been resolved by hypothesizing that adiponectin induces tolerance to inflammatory stimuli, notably Toll-like receptor (TLR) ligands. We noticed that this effect resembled lipopolysaccharide (LPS) tolerance and therefore tested adiponectin from a variety of sources for LPS contamination. All adiponectin tested carried low levels of LPS in the range of 1-30 pg/microg of adiponectin, sufficient to produce final LPS concentrations in the pg/ml range under experimental conditions. We found that induction of tolerance to TLR ligands by adiponectin in human monocyte-derived macrophages could be reproduced by such LPS concentrations. Moreover, the LPS antagonist, polymixin B, substantially inhibited induction of tolerance by adiponectin. Furthermore, polymixin B and a naturally occurring antagonist LPS were able to partially attenuate induction of tumour necrosis factor-alpha and interleukin-6 in human monocyte-derived macrophages by adiponectin. Polymixin B also inhibited nuclear factor-kappaB and mitogen-activated protein kinase signalling elicited by adiponectin. We therefore propose that some of adiponectin's immunomodulatory effects, in particular, its TLR-tolerising actions in human monocyte-derived macrophages, may be confounded by induction of tolerance by contaminating LPS. PMID- 19284498 TI - Activated T cells inhibit NK cell-mediated tumour rejection. AB - Previous studies have described the regulation of some T-cell subsets toward natural killer (NK) cells. Naturally occurring CD4(+)CD25(+) T regulatory cells can inhibit NK cell cytotoxicity, while activated interleukin-2 (IL-2) secreting T cells can stimulate NK cells. However, little is known about the impact of the integrity T-cell population on the final outcome of NK cell cytotoxicity. We thus examined the possible role of activated T cells in affecting NK cell cytotoxicity by mixed lymphocyte co-cultures in vitro and a B16 melanoma tumour model in vivo. In our study, activated T cells were found to be able to significantly inhibit NK cell cytotoxicity in vitro and blunt NK cell-mediated tumour rejection in vivo. The inhibition of NK cell function is a cell-cell contact dependent way. Results suggest that activated T cells may play an important role in limiting NK cell functions, which might be very significant for the design of biotherapy against tumour or infection in future. PMID- 19284499 TI - More vaccine efficacy studies on the recombinant Bacille Calmette-Guerin co expressing Ag85B, Mpt64 and Mtb8.4. AB - The immunogenicity of the recombinant Bacille Calmette-Guerin: rBCG-Ag85B Mpt64(190-198)-Mtb8.4 (rBCG-AMM) was evaluated in our previous study. This paper compares the protective efficacy of rBCG-AMM, rBCG-A which overexpresses Ag85B and BCG in C57BL/6 mice. There was no significant difference in proliferation characteristics among rBCG-AMM, rBCG-A and BCG. The growth characteristics of rBCG-AMM in host tissue were identical to control BCG, suggesting the improved protective efficacy was directly related to the expression of the Ag85B-Mpt64(190 198)-Mtb8.4 fusion protein. The protective experiment demonstrated that rBCG-AMM could confer similar or even better protective efficacy against Mycobacterium tuberculosis infection compared with BCG or rBCG-A as evaluated by bacterial organ loads, lung histopathology and net weight gain or loss. The results suggested that the recombinant BCG: rBCG-Ag85B-Mpt64(190-198)-Mtb8.4 is a potential vaccine candidate for further study. PMID- 19284500 TI - Differential activation of dendritic cells by Toll-like receptor agonists isolated from the Gram-positive vaccine vector Streptococcus gordonii. AB - The oral commensal bacterium Streptococcus gordonii has been gathering interest as a candidate live mucosal vaccine delivery vector. S. gordonii has been shown to be capable of activating antigen presenting immune cells in a manner which leads to their activation and maturation, yet the mechanism used by S. gordonii to do so is poorly understood. The aim of this work was to investigate the immunostimulatory components of S. gordonii in inducing murine dendritic cell (DC) activation and maturation. Lipoteichoic acid (LTA), lipoprotein (LP), peptidoglycan (PGN), and DNA were isolated from S. gordonii, and used to stimulate murine DC. Cytokine production and DC surface marker upregulation in response to the bacterial components was quantified by enzyme-linked immunosorbent assay and flow cytometry respectively. The results were contrasted against data obtained from DC derived from MyD88, TRIF [TIR(Toll/Interleukin-1 Receptor)-domain-containing adapter-inducing interferon-beta] or toll-like receptor-2 (TLR-2) knockout mice. The four S. gordonii bacterial components were found to differentially induce cytokine production and surface marker upregulation by murine DC. Activation of DC by both whole S. gordonii cells and the four bacterial components was abrogated in the absence of MyD88, but not in the absence of TRIF. LTA, LP and PGN, but not DNA and whole S. gordonii, required TLR-2 to induce a DC response. The results collectively indicate that S. gordonii activates DC predominantly through a MyD88-dependent and TRIF-independent pathway. This activation can be attributed to multiple immunostimulatory components present within S. gordonii bacterial cells. PMID- 19284501 TI - Transcriptional profiles in urine during acute rejection, bacteriuria, CMV infection and stable graft function after renal transplantation. AB - Acute rejection remains an important cause of renal allograft dysfunction and the need for accurate diagnosis is essential to treat transplant recipients successfully. Molecular markers in urine may serve as a diagnostic tool in acute rejection, but controversy still exists regarding the uniqueness of these biomarkers. We measured mRNA of perforin (PRF), granzyme B (GZMB) and granulysin (GNLY) normalized to cyclophilin B in urine specimens from 24 renal allograft recipients with acute rejection, 12 with bacteriuria, 11 with cytomegalovirus (CMV) infections and 17 controls with stable graft function. Measurements were performed using a real-time polymerase chain reaction assay. mRNA levels (means [95% CI]) for all three markers were significantly higher in recipients with acute rejection compared with controls: PRF (0.23 [0.12-0.42] versus 0.04 [0.02 0.07] P<0.001), GZMB (0.14 [0.09-0.23] versus 0.05 [0.03-0.08] P=0.003), GNLY (0.24 [0.14-0.41] versus 0.06 [0.03-0.11] P=0.001). GZMB and GNLY levels during acute rejection were significantly higher when compared with bacteriuria (P=0.011 and P=0.005 respectively), and PRF level during acute rejection was significantly elevated compared with CMV infection (P=0.015). No significant difference was found when comparing marker levels during bacteriuria and CMV infection to controls. Urinary mRNA levels of PRF, GZMB and GNLY are significantly elevated during acute rejection but not during bacteriuria or CMV infections when compared with recipients with stable graft function. The ability to differentiate acute rejection from bacteriuria and CMV infections was only present for some of the markers, that is why careful consideration should be given before applying this technique to clinical practice. PMID- 19284502 TI - Absence of tyrosinase-related protein-2/dopachrome tautomerase transcripts in PBMCs from vitiligo patients. AB - Tolerance is achieved by mechanisms occurring in both the thymus and periphery. Several reports have shown that presence of an antigen in the peripheral circulation results in tolerance induction. These reports imply that absence of a self-antigen can lead to induction of autoimmunity. Here, we show that tyrosinase related protein 2 (TRP-2) transcript is not detected in the peripheral blood mononuclear cells (PBMC) of vitiligo patients but is detected in healthy controls. Our result indicates that probably due to lack of expression in the PBMC, TRP-2 is not available for induction and maintenance of peripheral tolerance in vitiligo patients. It is also reported by others that co-stimulatory molecules are required for the initiation of autoimmune diseases in experimental models. We therefore analysed the transcript levels of these costimulatory molecules in vitiligo patients and healthy controls. We observed that the transcripts of B7.2 and CD40 molecules are more or less similar in both patients and controls. We could not detect B7.1 in any of the human subjects. Thus, we conclude that the antigen presenting cells (APC) are not in an activated state and that constitutively activated APC are possibly not required for the progression of the disease once it has been initiated. PMID- 19284503 TI - Complement inhibitor C4b-binding protein in primary Sjogren's syndrome and its association with other disease markers. AB - A subgroup of patients suffering from primary Sjogren's syndrome (pSS) display unexplained low levels of complement components C3 and/or C4 which is associated with increased risk of non-Hodgkin's lymphoma. C4b-binding protein (C4BP) is a major fluid-phase complement inhibitor which can influence C4 and C3 levels. Therefore we analysed C4BP levels in the sera of patients with pSS to better understand the disturbances in complement in pSS. Associations with other disease markers were also investigated to define a possible role of C4BP as marker of high-risk disease course. Plasma levels of C4BP were analysed in pSS patients (n=86) and in controls (n=68) by ELISA. C4BP levels from 49 patients were correlated to disease activity markers and autoantibody profiles. We found that total C4BP plasma levels were significantly higher in pSS patients compared with controls. C4BP levels correlated to the acute phase response, to levels of C4 and C3 as well as to the CD4+/CD8+ T-cell ratio. C4BP levels were inversely related to IgG levels, extent of autoantibody production and global disease activity. C3dg levels, a marker of complement activation, displayed a negative correlation to C4 levels but interestingly not to C4BP levels. In conclusion, C4BP levels are increased in patients suffering from pSS proportional to their acute phase response. However, in the most active cases, with the most widespread autoantibody production, C4BP levels were decreased in parallel with levels of C3 and C4 and CD4+ T cells, suggesting that disturbed complement regulation may contribute to pathogenicity in pSS. PMID- 19284504 TI - Eosinophil cationic protein stimulates migration of human lung fibroblasts in vitro. AB - Asthma is characterized by eosinophilic inflammation and remodelling of the airways. Eosinophil cationic protein (ECP) is a protein released by activated eosinophils and the hypothesis that ECP contributes to the development of structural changes in the airways of asthmatics has been posed. Fibroblast recruitment is an important step in the remodelling process, and we therefore put the question whether ECP stimulates migration of human lung fibroblasts. Human peripheral eosinophils isolated from buffycoats from healthy individuals were cultured and conditioned media (CM) were collected. Native ECP was extracted from human peripheral eosinophils by gel filtration, ion-exchange and chelating chromatography. The ability of eosinophil CM and ECP to stimulate fibroblast migration was determined using the 48-well Boyden chamber. ECP concentrations in CM were assayed by ECP-CAP-FEIA. Both CM and ECP significantly stimulated fibroblast migration (48.4+/-cells/field versus 33+/-2 and 36+/-6 versus 25+/-4; P<0.001 and 0.05 respectively) in a time- and concentration-dependent manner. Adding neutralizing ECP antibodies attenuated fibroblast migration induced by both ECP as well as CM. ECP stimulates migration of human lung fibroblasts, suggesting a potential mechanism for eosinophils in the fibrotic response. This may be an important mechanism by which ECP promotes remodelling of extracellular matrix leading to airway fibrosis in asthmatics. PMID- 19284505 TI - Oral health conditions in Italian Special Olympics athletes. AB - During three Italian Special Olympics National Games, 365 athletes were screened. Dental and medical conditions and demographic data were recorded. The athletes were divided into two groups: those with Down syndrome (DS) and those without DS but who had intellectual disabilities (non-DS). Most of the subjects were in good systemic health. Total DMFt was 10.3 (SD 5.8; D=1.3; M=6.1; F=2.8). Decayed and filled teeth were significantly more frequent in athletes who did not have DS compared to those with DS. No significant differences were found between the two groups in the number of subjects with filled, sealed, or traumatized teeth. Athletes with DS and without DS who participated in the Italian Special Olympics had a similar oral status, which was better than Italian persons who were institutionalized and who had an intellectual disability. PMID- 19284506 TI - Radiation-associated carotid artery atherosclerosis: case report and review of contemporaneous literature. AB - Radiation therapy (RT) is a component of the treatment of patients with head and neck malignancies. This therapy may damage the nearby carotid arteries, thereby initiating or accelerating the atherosclerotic process (atheroma formation). Dentists treating patients who have been irradiated should examine the patient's panoramic radiograph for evidence of atheroma-like calcifications, which appear 1.5 to 2.5 cm posterior and inferior to the angle of the mandible. Patients with evidence of such lesions should be referred to their primary care physician with the suggestion that an ultrasound examination of the carotid arteries is indicated. PMID- 19284507 TI - Antigen-presenting cells in human immunosuppressive drug-induced gingival enlargement. AB - An immunoperoxidase technique was used to compare the number of CD1a+ and factor XIIIa+ dendritic cells (DCs), and CD68+ Macrophages (M) in 30 gingival samples from subjects with clinically healthy periodontitium (HP) and 10 samples from subjects with drug-induced gingival enlargement (DIGE). Fewer CD1a+ and factor XIIIa+ DCs were found in areas with inflammatory infiltration (II) of the lamina propria (LP) in the group with immunosuppressed DIGE (IDIGE) compared to the group with HP. In the sulcular and junctional/pocket epithelia, the number of CD1a+ DCs was decreased in the group with IDIGE (p<0.05). There was a tendency toward a reduced number of CD1a+ DCs and CD68+ M in areas without inflammatory infiltrate of the LP in the group with IDIGE. The alterations in the number of antigen-presenting cells (APCs) may be the reason for the decreased periodontal inflammation and breakdown clinically observed in subjects who are immunosuppressed. PMID- 19284508 TI - Toothache pain: behavioral impact and self-care strategies. AB - A computer-assisted telephone interview in Maryland of adults who had low income and were Hispanic, Black, and White and who had experienced a toothache during the previous 12 months was conducted. Respondents reported a high prevalence of toothaches, with 44.3% having experienced more than five toothaches during the preceding 10 years. Pain intensity associated with the most recent toothache was high with 45.1% of the respondents reporting the highest pain possible. Pain interfered with many aspects of normal functioning. Self-care strategies generally took precedence over professional health services. Pain sufferers used a combination of self-care and formal care strategies. Initial strategies most often focused on nonprescription medicines(home remedies and prayer. The majority of respondents ultimately sought pain relief from a dentist. We identified a number of significant differences in the strategies used across racial/ethnic groups. PMID- 19284509 TI - Correlation between mothers' perceptions/expectations and the behavior of children with HIV during dental treatment. AB - The aim of this study was to evaluate the mothers' perceptions and expectations of the behavior of their children who were HIV positive, during dental treatment. The sample was composed of 40 mother and child pairs. The mothers answered seven questions during two different appointments (M1, the child's initial appointment, and M2, the child's treatment appointment) in order to identify their perceptions and expectations about their children's behavior during dental treatment. In addition, child's behavior during the appointments was evaluated. The results of the mothers' interviews were then correlated with their children's behavior using Spearman's correlation test (p<0.05). There was no significant correlation between the mothers' perceptions and expectations and the behavior displayed by their children during dental treatment (p>0.05). PMID- 19284510 TI - Oral findings in adults with osteogenesis imperfecta. AB - This paper describes oral findings in an adult population with osteogenesis imperfecta (OI) in Norway (n=94). All participants underwent a structured interview and an oral examination. Panoramic radiographs were analyzed. The findings were compared with data from other Nordic epidemiological studies. Seventeen individuals (19%) had clinical signs of dentinogenesis imperfecta (DI). Persons with OI had twice as many missing teeth as the general population, and the number of endodontically treated teeth was higher than in the general population. All persons in the study had an acceptable state of tooth tissue loss. Almost all (97%) of the participants in the study group visited a dentist or a dental hygienist regularly. The results of the study indicated a low prevalence of clinical DI compared with previous reported studies. Oral health was not as good in the population with OI when compared with the general population, although daily oral health habits were good and dental visits were regular. PMID- 19284514 TI - Lamivudine combined with hepatitis B immunoglobulin in for prophylaxis of hepatitis B recurrence after liver transplantation: time for a change? PMID- 19284518 TI - Large-scale association study for structural soundness and leg locomotion traits in the pig. AB - BACKGROUND: Identification and culling of replacement gilts with poor skeletal conformation and feet and leg (FL) unsoundness is an approach used to reduce sow culling and mortality rates in breeding stock. Few candidate genes related to soundness traits have been identified in the pig. METHODS: In this study, 2066 commercial females were scored for 17 traits describing body conformation and FL structure, and were used for association analyses. Genotyping of 121 SNPs derived from 95 genes was implemented using Sequenom's MassARRAY system. RESULTS: Based on the association results from single trait and principal components using mixed linear model analyses and false discovery rate testing, it was observed that APOE, BMP8, CALCR, COL1A2, COL9A1, DKFZ, FBN1 and VDBP were very highly significantly (P < 0.001) associated with body conformation traits. The genes ALOX5, BMP8, CALCR, OPG, OXTR and WNT16 were very highly significantly (P < 0.001) associated with FL structures, and APOE, CALCR, COL1A2, GNRHR, IHH, MTHFR and WNT16 were highly significantly (P < 0.01) associated with overall leg action. Strong linkage disequilibrium between CALCR and COL1A2 on SSC9 was detected, and haplotype -ACGACC- was highly significantly (P < 0.01) associated with overall leg action and several important FL soundness traits. CONCLUSION: The present findings provide a comprehensive list of candidate genes for further use in fine mapping and biological functional analyses. PMID- 19284519 TI - Detecting selection-induced departures from Hardy-Weinberg proportions. AB - Viability selection influences the genotypic contexts of alleles and leads to quantifiable departures from Hardy-Weinberg proportions. One measure of these departures is Wright's inbreeding coefficient (F), where observed heterozygosity is compared with expected heterozygosity. Here, I extend population genetics theory to describe post-selection genotype frequencies in terms of post-selection allele frequencies and fitness dominance. The resulting equations correspond to non-equilibrium populations, allowing the following questions to be addressed: When selection is present, how large a sample size is needed to detect significant departures from Hardy-Weinberg? How do selection-induced departures from Hardy-Weinberg vary with allele frequencies and levels of fitness dominance? For realistic selection coefficients, large sample sizes are required and departures from Hardy-Weinberg proportions are small. PMID- 19284520 TI - Factor-analytic models for genotype x environment type problems and structured covariance matrices. AB - BACKGROUND: Analysis of data on genotypes with different expression in different environments is a classic problem in quantitative genetics. A review of models for data with genotype x environment interactions and related problems is given, linking early, analysis of variance based formulations to their modern, mixed model counterparts. RESULTS: It is shown that models developed for the analysis of multi-environment trials in plant breeding are directly applicable in animal breeding. In particular, the 'additive main effect, multiplicative interaction' models accommodate heterogeneity of variance and are characterised by a factor analytic covariance structure. While this can be implemented in mixed models by imposing such structure on the genetic covariance matrix in a standard, multi trait model, an equivalent model is obtained by fitting the common and specific factors genetic separately. Properties of the mixed model equations for alternative implementations of factor-analytic models are discussed, and extensions to structured modelling of covariance matrices for multi-trait, multi environment scenarios are described. CONCLUSION: Factor analytic models provide a natural framework for modelling genotype x environment interaction type problems. Mixed model analyses fitting such models are likely to see increasing use due to the parsimonious description of covariance structures available, the scope for direct interpretation of factors as well as computational advantages. PMID- 19284521 TI - A biologically active peptide mimetic of N-acetylgalactosamine/galactose. AB - BACKGROUND: Glycosylated proteins and lipids are important regulatory factors whose functions can be altered by addition or removal of sugars to the glycan structure. The glycans are recognized by sugar-binding lectins that serve as receptors on the surface of many cells and facilitate initiation of an intracellular signal that changes the properties of the cells. We identified a peptide that mimics the ligand of an N-acetylgalactosamine (GalNAc)-specific lectin and asked whether the peptide would express specific biological activity. FINDINGS: A 12-mer phage display library was screened with a GalNAc-specific lectin to identify an amino acid sequence that binds to the lectin. Phage particles that were eluted from the lectin with free GalNAc were considered to have been bound to a GalNAc-binding site. Peptides were synthesized with the selected sequence as a quadravalent structure to facilitate receptor crosslinking. Treatment of human peripheral blood mononuclear cells for 24 h with the peptide stimulated secretion of interleukin-8 (IL-8) but not of IL-1beta, IL 6, IL-10, or tumor necrosis factor-alpha (TNF-alpha). The secretion of IL-21 was stimulated as strongly with the peptide as with interferon-gamma. CONCLUSION: The data indicate that the quadravalent peptide has biological activity with a degree of specificity. These effects occurred at concentrations in the nanomolar range, in contrast to free sugars that generally bind to proteins in the micro- to millimolar range. PMID- 19284522 TI - Analysis of apyrase 5' upstream region validates improved Anopheles gambiae transformation technique. AB - BACKGROUND: Genetic transformation of the malaria mosquito Anopheles gambiae has been successfully achieved in recent years, and represents a potentially powerful tool for researchers. Tissue-, stage- and sex-specific promoters are essential requirements to support the development of new applications for the transformation technique and potential malaria control strategies. During the Plasmodium lifecycle in the invertebrate host, four major mosquito cell types are involved in interactions with the parasite: hemocytes and fat body cells, which provide humoral and cellular components of the innate immune response, midgut and salivary glands representing the epithelial barriers traversed by the parasite during its lifecycle in the mosquito. FINDINGS: We have analyzed the upstream regulatory sequence of the An. gambiae salivary gland-specific apyrase (AgApy) gene in transgenic An. gambiae using a piggyBac transposable element vector marked by a 3xP3 promoter:DsRed gene fusion. Efficient germ-line transformation in An. gambiae mosquitoes was obtained and several integration events in at least three different G0 families were detected. LacZ reporter gene expression was analyzed in three transgenic lines/groups, and in only one group was tissue specific expression restricted to salivary glands. CONCLUSION: Our data describe an efficient genetic transformation of An. gambiae embryos. However, expression from the selected region of the AgApy promoter is weak and position effects may mask tissue- and stage- specific activity in transgenic mosquitoes. PMID- 19284523 TI - Cutaneous symptoms such as acneform eruption and pigmentation are closely associated with blood levels of 2,3,4,7,8-penta-chlorodibenzofurans in Yusho patients, using data mining analysis. AB - BACKGROUND: Yusho an intoxication caused by oral dioxins and polychlorinated biphenyls occurred in 1968. Patients suffered from various systemic symptoms, including general fatigue, nausea, muscular and articular pain, acneform eruptions, black comedones, cutaneous and oral pigmentation, and increased eye discharge. The major causative factor was the contamination of rice oil with 2,3,4,7,8-penta-chlorodibenzofuran (PeCDF). Recent technical advances have allowed us to measure blood levels of PeCDF. However, there is little information on which symptoms and laboratory data are directly associated with PeCDF levels. METHODS: Yusho patients underwent annual medical check-ups from 2001 to 2003. Blood PeCDF levels were correlated with the presence or absence of symptoms in medical, hematological, dermatological, dental and ophthalmological examinations. This study analyzed all combinations by using the association analysis. This is the most suitable method to evaluate all combinations of the data comprehensively. This method was used to determine the rate of patients with high PeCDF level in the population with each symptom, and to extract combinations of three symptoms which were strongly associated with high PeCDF level. RESULTS AND CONCLUSION: The rate of the patients with high PeCDF level was high in populations with high uric acid, black comedones (face), second highest quartile of age, or high urea nitrogen. The combination of three symptoms associated with the highest rate of patients with high PeCDF level was "high uric acid, female sexuality, and history of acneform eruptions", followed by "history of Yusho in and after 1968, high cholesterol level, and subjective symptoms." This analysis newly suggested that PeCDF concentration may be associated with history of dermatological symptoms, high uric acid, and elevated erythrocyte sedimentation rate. PMID- 19284524 TI - Markers typed in genome-wide analysis identify regions showing deviation from Hardy-Weinberg equilibrium. AB - BACKGROUND: Deviations from Hardy-Weinberg equilibrium (HWE) are commonly thought of as indicating genotyping errors, population stratification or some other artefact. However they could also arise through important biological mechanisms. In particular, genetic variants having a recessive effect on the successful fertilisation and/or development of an embryo might be manifest through such deviations in an unselected sample of "control" subjects. FINDINGS: We investigated genotypes from 463842 autosomal markers from 1504 British subjects. We identified regions in which several neighbouring markers exhibited deviation from HWE in the same direction by considering "heterozygosity scores" in windows of 10 markers. The heterozygosity score for each marker was defined as -log(p) or log(p) according to whether the marker demonstrated increased heterozygosity or homozygosity. In each window the marker with the highest absolute score was ignored and the positive and negative scores were summed for the other nine markers. Windows were selected on the basis of this sum exceeding a given threshold, for which we used values of 50 or 15.For the threshold of 50, we identified 7 regions with increased heterozygosity and for the threshold of 15 we identified 22 regions with increased heterozygosity, 23 with increased homozygosity and 2 containing both kinds of window. The most impressive of these results came from a group of 6 markers at 17q21, each of which showed increased heterozygosity significant at p < 10(-190). CONCLUSION: The human genome contains regions which deviate markedly from HWE and these might harbour genes influencing embryonic survival. PMID- 19284525 TI - Current practices in the use of sildenafil for pulmonary arterial hypertension in Brazilian hospitals. AB - BACKGROUND: Sildenafil is a cyclic guanosine monophosphate-specific phosphodiesterase-5 inhibitor used for treating pulmonary hypertension. Although the use of sildenafil in patients under 18 years old is off-label, this inhibitor has been widely prescribed for children treatment at hospitals in Brazil. In this work we evaluated the current practices in using sildenafil in the three main reference hospitals of Rio de Janeiro to design a clinical trial. Then we analyzed the content of sildenafil in powder paper preparations used in these institutions. METHODS AND RESULTS: We assessed the data about the use of sildenafil in three reference hospitals including Instituto Nacional de Cardiologia - INC, Instituto Estadual de Cardiologia Aloysio de Castro - IECAC and Hospital Pro-Cardiaco - HPC. The pharmacy records were analyzed from April 1st, 2008 to April 30th, 2008 and interviews with the pharmacists were also performed. Our results showed that INC used the greatest amount of sildenafil in: treatment of pulmonary arterial hypertension (PAH), management of transient PAH during surgery, preparation for cardiac transplantation and haemodynamic studies during cardiac catheterization. Meanwhile IECAC and HPC used sildenafil only for treating PAH in few patients during the period evaluated. In INC and IECAC, sildenafil was available in tablets, and powder papers prepared by two private pharmacies and one public hospital pharmacy. In contrast all patients of HPC received sildenafil in tablets with no external manipulation. Our quantification analysis results using reverse-phase high performance liquid chromatography method showed that powder papers prepared by the private pharmacies from the sildenafil tablets presented only 58.5 to 89.3% of the declared concentration in contrast to samples from the public hospital pharmacy (104.4 to 105.3%). CONCLUSION: Few patients received the prescribed sildenafil dose at the reference hospitals evaluated in Rio de Janeiro, which may importantly compromise this inhibitor effect in the current treatment. This study reinforced the need of checking the practices of preparing and administering sildenafil continually. PMID- 19284526 TI - Overview of current and future biologically based targeted therapies in head and neck squamous cell carcinoma. AB - Recent advances in genomics, proteomics, bioinformatics and systems biology have unraveled the complex aberrant signaling networks in cancer. The knowledge accrued has dramatically increased the opportunities for discovery of novel molecular targets for drug development. Major emphasis is being laid on designing new therapeutic strategies targeting multiple signaling pathways for more effective disease management. However, the translation of in vitro findings to patient management often poses major challenges that limit their clinical efficacy. Here we will discuss how understanding the dysregulated signaling networks can explain the pitfalls in translating the laboratory findings from the bench-to-bedside and suggest novel approaches to overcome these problems using head and neck cancer as a prototype. The five year survival rates of HNSCC patients (about 50% at 5 years) have not improved significantly despite advancements in multimodality therapy including surgery, radiation and chemotherapy. Molecular targeted therapies with inhibitors of EGFR and VEGF either alone, or in combination with conventional treatments have shown limited improved efficacy. The key deregulated signaling pathways in head and neck squamous cell carcinoma (HNSCC) include EGFR, Ras, TGFbeta, NFkappaB, Stat, Wnt/beta-catenin and PI3-K/AKT/mTOR. The aberrant activities of these interrelated signaling pathways contribute to HNSCC development. In depth understanding of the cross-talks between these pathways and networks will form the basis of developing novel strategies for targeting multiple molecular components for more effective prevention and treatment of HNSCC. PMID- 19284527 TI - Rotational strength, range of motion, and function in people with unaffected shoulders from various stages of life. AB - BACKGROUND: Different measurements are used to assess shoulder function, including range of motion, strength, functional performance and self-report function. To understand disablement, it is necessary to understand the relationship between impairments and function in persons without shoulder problems. This study was conducted to enhance existing comparative data in subjects without upper extremity pathology, and to assess the relationships between impairments (range of motion, strength) and self-reported or measured function/disability. The impact of age, gender and dominance was determined. METHODS: Two-hundred ninety-four subjects with unaffected shoulders were recruited. The subjects (mean age: 37 years old) were divided into three subgroups, 18-39, 40-59, and over 60 years of age. During a single session, at least two of the following variables were measured: self-reported function (shoulder disability scales), range of motion, isometric rotational strength, or upper limb functional performance (FIT-HaNSA). Two-way analysis of variance was used to determine, for each variable, the effects of age and gender. The relationship between the outcomes was established using Pearson product correlations. RESULTS: Men were significantly stronger than women for all age categories. There was an age-related decline in strength in men in the over-60 age category. Significant negative correlations between strength and range of motion were demonstrated (-0.22 99% DNA sequence identity to that of Tn501. The mer operon was present on a transposon, designated TnAO22, captured by introducing a broad-host-range IncP plasmid into Achromobacter sp. AO22 and subsequently transferring it to E. coli recipients. The transposition frequency of TnAO22 was 10-2 to 10-3 per target plasmid transferred. Analysis of TnAO22 sequence revealed it belonged to the Tn21 subgroup of the Tn3 superfamily of transposons, with the transposition module having >99% identity with Tn5051 of a Pseudomonas putida strain isolated from a water sample in New York. CONCLUSION: TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium. Achromobacter sp. AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes. PMID- 19284536 TI - Early biomarkers and potential mediators of ventilation-induced lung injury in very preterm lambs. AB - BACKGROUND: Bronchopulmonary dysplasia (BPD) is closely associated with ventilator-induced lung injury (VILI) in very preterm infants. The greatest risk of VILI may be in the immediate period after birth, when the lungs are surfactant deficient, still partially filled with liquid and not uniformly aerated. However, there have been very few studies that have examined this immediate post-birth period and identified the initial injury-related pathways that are activated. We aimed to determine if the early response genes; connective tissue growth factor (CTGF), cysteine rich-61 (CYR61) and early growth response 1 (EGR1), were rapidly induced by VILI in preterm lambs and whether ventilation with different tidal volumes caused different inflammatory cytokine and early response gene expression. METHODS: To identify early markers of VILI, preterm lambs (132 d gestational age; GA, term approximately 147 d) were resuscitated with an injurious ventilation strategy (V(T) 20 mL/kg for 15 min) then gently ventilated (5 mL/kg) for 15, 30, 60 or 120 min (n = 4 in each). To determine if early response genes and inflammatory cytokines were differentially regulated by different ventilation strategies, separate groups of preterm lambs (125 d GA; n = 5 in each) were ventilated from birth with a V(T) of 5 (VG5) or 10 mL/kg (VG10) for 135 minutes. Lung gene expression levels were compared to levels prior to ventilation in age-matched control fetuses. RESULTS: CTGF, CYR61 and EGR1 lung mRNA levels were increased approximately 25, 50 and 120-fold respectively (p < 0.05), within 30 minutes of injurious ventilation. VG5 and VG10 caused significant increases in CTGF, CYR61, EGR1, IL1- , IL-6 and IL-8 mRNA levels compared to control levels. CTGF, CYR61, IL-6 and IL-8 expression levels were higher in VG10 than VG5 lambs; although only the IL-6 and CYR61 mRNA levels reached significance. CONCLUSION: CTGF, CYR61 and EGR1 may be novel early markers of lung injury and mechanical ventilation from birth using relatively low tidal volumes may be less injurious than using higher tidal volumes. PMID- 19284538 TI - Bisulfite-based epityping on pooled genomic DNA provides an accurate estimate of average group DNA methylation. AB - BACKGROUND: DNA methylation plays a vital role in normal cellular function, with aberrant methylation signatures being implicated in a growing number of human pathologies and complex human traits. Methods based on the modification of genomic DNA with sodium bisulfite are considered the 'gold-standard' for DNA methylation profiling on genomic DNA; however, they require relatively large amounts of DNA and may be prohibitively expensive when used on the large sample sizes necessary to detect small effects. We propose that a high-throughput DNA pooling approach will facilitate the use of emerging methylomic profiling techniques in large samples. RESULTS: Compared with data generated from 89 individual samples, our analysis of 205 CpG sites spanning nine independent regions of the genome demonstrates that DNA pools can be used to provide an accurate and reliable quantitative estimate of average group DNA methylation. Comparison of data generated from the pooled DNA samples with results averaged across the individual samples comprising each pool revealed highly significant correlations for individual CpG sites across all nine regions, with an average overall correlation across all regions and pools of 0.95 (95% bootstrapped confidence intervals: 0.94 to 0.96). CONCLUSION: In this study we demonstrate the validity of using pooled DNA samples to accurately assess group DNA methylation averages. Such an approach can be readily applied to the assessment of disease phenotypes reducing the time, cost and amount of DNA starting material required for large-scale epigenetic analyses. PMID- 19284537 TI - The erratic mitochondrial clock: variations of mutation rate, not population size, affect mtDNA diversity across birds and mammals. AB - BACKGROUND: During the last ten years, major advances have been made in characterizing and understanding the evolution of mitochondrial DNA, the most popular marker of molecular biodiversity. Several important results were recently reported using mammals as model organisms, including (i) the absence of relationship between mitochondrial DNA diversity and life-history or ecological variables, (ii) the absence of prominent adaptive selection, contrary to what was found in invertebrates, and (iii) the unexpectedly large variation in neutral substitution rate among lineages, revealing a possible link with species maximal longevity. We propose to challenge these results thanks to the bird/mammal comparison. Direct estimates of population size are available in birds, and this group presents striking life-history trait differences with mammals (higher mass specific metabolic rate and longevity). These properties make birds the ideal model to directly test for population size effects, and to discriminate between competing hypotheses about the causes of substitution rate variation. RESULTS: A phylogenetic analysis of cytochrome b third-codon position confirms that the mitochondrial DNA mutation rate is quite variable in birds, passerines being the fastest evolving order. On average, mitochondrial DNA evolves slower in birds than in mammals of similar body size. This result is in agreement with the longevity hypothesis, and contradicts the hypothesis of a metabolic rate dependent mutation rate. Birds show no footprint of adaptive selection on cytochrome b evolutionary patterns, but no link between direct estimates of population size and cytochrome b diversity. The mutation rate is the best predictor we have of within-species mitochondrial diversity in birds. It partly explains the differences in mitochondrial DNA diversity patterns observed between mammals and birds, previously interpreted as reflecting Hill-Robertson interferences with the W chromosome. CONCLUSION: Mitochondrial DNA diversity patterns in birds are strongly influenced by the wide, unexpected variation of mutation rate across species. From a fundamental point of view, these results are strongly consistent with a relationship between species maximal longevity and mitochondrial mutation rate, in agreement with the mitochondrial theory of ageing. Form an applied point of view, this study reinforces and extends the message of caution previously expressed for mammals: mitochondrial data tell nothing about species population sizes, and strongly depart the molecular clock assumption. PMID- 19284539 TI - How to select a chiropractor for the management of athletic conditions. AB - BACKGROUND: Chiropractors are an integral part of the management of musculoskeletal injuries. A considerable communication gap between the chiropractic and medical professions exists. Subsequently referring allopathic practitioners lack confidence in picking a chiropractic practitioner with appropriate management strategies to adequately resolve sporting injuries. Subsequently, the question is often raised: "how do you find a good chiropractor?". DISCUSSION: Best practice guidelines are increasingly suggesting that musculoskeletal injuries should be managed with multimodal active and passive care strategies. Broadly speaking chiropractors may be subdivided into "modern multimodal" or "classical" (unimodal) in nature. The modern multimodal practitioner is better suited to managing sporting injuries by incorporating passive and active care management strategies to address three important phases of care in the continuum of injury from the acute inflammation/pain phase to the chronic/rehabilitation phase to the injury prevention phase. In contrast, the unimodal, manipulation only and typically spine only approach of the classical practitioner seems less suited to the challenges of the injured athlete. Identifying what part of the philosophical management spectrum a chiropractor falls is important as it is clearly not easily evident in most published material such as Yellow Pages advertisements. SUMMARY: Identifying a chiropractic practitioner who uses multimodal treatment of adequate duration, who incorporates active and passive components of therapy including exercise prescription whilst using medical terminology and diagnosis without mandatory x-rays or predetermined treatment schedules or prepaid contracts of care will likely result in selection of a chiropractor with the approach and philosophy suited to appropriately managing athletic conditions. Sporting organizations and associations should consider using similar criteria as a minimum standard to allow participation in health care team selections. PMID- 19284540 TI - Recent developments in StemBase: a tool to study gene expression in human and murine stem cells. AB - BACKGROUND: Currently one of the largest online repositories for human and mouse stem cell gene expression data, StemBase was first designed as a simple web interface to DNA microarray data generated by the Canadian Stem Cell Network to facilitate the discovery of gene functions relevant to stem cell control and differentiation. FINDINGS: Since its creation, StemBase has grown in both size and scope into a system with analysis tools that examine either the whole database at once, or slices of data, based on tissue type, cell type or gene of interest. As of September 1, 2008, StemBase contains gene expression data (microarray and Serial Analysis of Gene Expression) from 210 stem cell samples in 60 different experiments. CONCLUSION: StemBase can be used to study gene expression in human and murine stem cells and is available at http://www.stembase.ca. PMID- 19284541 TI - Finding evolutionarily conserved cis-regulatory modules with a universal set of motifs. AB - BACKGROUND: Finding functional regulatory elements in DNA sequences is a very important problem in computational biology and providing a reliable algorithm for this task would be a major step towards understanding regulatory mechanisms on genome-wide scale. Major obstacles in this respect are that the fact that the amount of non-coding DNA is vast, and that the methods for predicting functional transcription factor binding sites tend to produce results with a high percentage of false positives. This makes the problem of finding regions significantly enriched in binding sites difficult. RESULTS: We develop a novel method for predicting regulatory regions in DNA sequences, which is designed to exploit the evolutionary conservation of regulatory elements between species without assuming that the order of motifs is preserved across species. We have implemented our method and tested its predictive abilities on various datasets from different organisms. CONCLUSION: We show that our approach enables us to find a majority of the known CRMs using only sequence information from different species together with currently publicly available motif data. Also, our method is robust enough to perform well in predicting CRMs, despite differences in tissue specificity and even across species, provided that the evolutionary distances between compared species do not change substantially. The complexity of the proposed algorithm is polynomial, and the observed running times show that it may be readily applied. PMID- 19284542 TI - Outcomes of resection and non-resection strategies in management of patients with advanced colorectal cancer. AB - BACKGROUND: The management of patients with surgically incurable bowel cancer at presentation is controversial. The aims of treatment are to optimise quality of life and prolong survival. It has been believed that the most effective palliation is achieved by resection of the primary cancer in order to pre-empt future complications. This study reviews and compares the outcomes of patients with incurable bowel cancer managed by resection and non-resection strategies over a 7-year period in a single District General Hospital. PATIENTS AND METHODS: All patients with surgically incurable bowel cancer at presentation were identified from the prospectively collected local ACPGBI database. Survival, using Kaplan-Meier method and log-rank test, was compared between patients managed by resection of the primary, non-resectional intervention (surgery, stent & oncological treatments) and those managed with supportive care only. The primary endpoint of the study was survival on an intention to treat basis, compared using Kaplan-Meier and log-rank tests. RESULTS: Of 646 consecutive newly diagnosed bowel cancer patients over a 7 year period 154 cases (24%) were deemed surgically incurable at presentation. Of these surgical resection was carried out in 45 patients (29%), non-resectional intervention was followed in 52 patients (34%) and supportive treatment alone in 57 patients (37%). Median survival of each group was as follows: resected patients 11 months (I.Q range 3-18 months), non-resectional intervention 7 months (I.Q range 2-15 months) and supportive care alone 2 months (I.Q range 1-8 months). Only one patient (2%) managed by non resectional intervention required later surgery to treat primary tumour related complications. Survival was not significantly different between resection and non resection treatments. The overall operative mortality for the resection group was 16% (7/45 cases), with an elective mortality of 14% (4/28 cases) and emergency mortality 18% (3/17 cases). CONCLUSION: In an unselected bowel cancer population surgical resection of the primary tumour in patients presenting with incurable disease does not improve survival and is associated with a high risk of post operative mortality. PMID- 19284543 TI - Towards a basic endoscopic evaluation of swallowing in acute stroke - identification of salient findings by the inexperienced examiner. AB - BACKGROUND: Dysphagia is common after stroke. Fiberoptic endoscopic evaluation of swallowing (FEES) is a powerful tool for dysphagia assessment. The purpose of this study was to assess whether a previously established endoscopic examination protocol based on the identification of typical findings indicative of stroke - related dysphagia may be learned and adopted by clinicians so far inexperienced in this field. METHODS: After receiving a structured lecture on this topic, participants were asked to rate video sequences of endoscopic swallowing examinations of acute stroke patients. The first part of the testing ("single findings-rating") comprised of 16 single sequences, the second part ("complete examination-rating") presented the key sequences of 8 complete examinations. Before the second part was started, results of the first were discussed. RESULTS: At the "single findings-rating" 88.8% of video-sequences were assessed correctly, while at the "complete examination-rating" the average performance had improved to 96%. Furthermore, no overlooking of relevant pathologies was noted in the second part of the testing. CONCLUSION: This study suggests that the presented endoscopic examination protocol is reliably interpreted by inexperienced clinicians after a short lecture and may therefore easily and successfully be adopted in dysphagia management of acute stroke care. PMID- 19284544 TI - Aphids acquired symbiotic genes via lateral gene transfer. AB - BACKGROUND: Aphids possess bacteriocytes, which are cells specifically differentiated to harbour the obligate mutualist Buchnera aphidicola (gamma Proteobacteria). Buchnera has lost many of the genes that appear to be essential for bacterial life. From the bacteriocyte of the pea aphid Acyrthosiphon pisum, we previously identified two clusters of expressed sequence tags that display similarity only to bacterial genes. Southern blot analysis demonstrated that they are encoded in the aphid genome. In this study, in order to assess the possibility of lateral gene transfer, we determined the full-length sequences of these transcripts, and performed detailed structural and phylogenetic analyses. We further examined their expression levels in the bacteriocyte using real-time quantitative RT-PCR. RESULTS: Sequence similarity searches demonstrated that these fully sequenced transcripts are significantly similar to the bacterial genes ldcA (product, LD-carboxypeptidase) and rlpA (product, rare lipoprotein A), respectively. Buchnera lacks these genes, whereas many other bacteria, including Escherichia coli, a close relative of Buchnera, possess both ldcA and rlpA. Molecular phylogenetic analysis clearly demonstrated that the aphid ldcA was derived from a rickettsial bacterium closely related to the extant Wolbachia spp. (alpha-Proteobacteria, Rickettsiales), which are intracellular symbionts of various lineages of arthropods. The evolutionary origin of rlpA was not fully resolved, but it was clearly demonstrated that its double-psi beta-barrel domain is of bacterial origin. Real-time quantitative RT-PCR demonstrated that ldcA and rlpA are expressed 11.6 and 154-fold higher in the bacteriocyte than in the whole body, respectively. LdcA is an enzyme required for recycling murein (peptidoglycan), which is a component of the bacterial cell wall. As Buchnera possesses a cell wall composed of murein but lacks ldcA, a high level of expression of the aphid ldcA in the bacteriocyte may be essential to maintain Buchnera. Although the function of RlpA is not well known, conspicuous up regulation of the aphid rlpA in the bacteriocyte implies that this gene is also essential for Buchnera. CONCLUSION: In this study, we obtained several lines of evidence indicating that aphids acquired genes from bacteria via lateral gene transfer and that these genes are used to maintain the obligately mutualistic bacterium, Buchnera. PMID- 19284545 TI - A gene-rich linkage map in the dioecious species Actinidia chinensis (kiwifruit) reveals putative X/Y sex-determining chromosomes. AB - BACKGROUND: The genus Actinidia (kiwifruit) consists of woody, scrambling vines, native to China, and only recently propagated as a commercial crop. All species described are dioecious, but the genetic mechanism for sex-determination is unknown, as is the genetic basis for many of the cluster of characteristics making up the unique fruit. It is, however, an important crop in the New Zealand economy, and a classical breeding program would benefit greatly by knowledge of the trait alleles carried by both female and male parents. The application of marker assisted selection (MAS) in seedling populations would also aid the accurate and efficient development of novel fruit types for the market. RESULTS: Gene-rich female, male and consensus linkage maps of the diploid species A. chinensis have been constructed with 644 microsatellite markers. The maps consist of twenty-nine linkage groups corresponding to the haploid number n = 29. We found that sex-linked sequence characterized amplified region (SCAR) markers and the 'Flower-sex' phenotype consistently mapped to a single linkage group, in a subtelomeric region, in a section of inconsistent marker order. The region also contained markers of expressed genes, some of unknown function. Recombination, assessed by allelic distribution and marker order stability, was, in the remainder of the linkage group, in accordance with other linkage groups. Fully informative markers to other genes in this linkage group identified the comparative linkage group in the female map, where recombination ratios determining marker order were similar to the autosomes. CONCLUSION: We have created genetic linkage maps that define the 29 linkage groups of the haploid genome, and have revealed the position and extent of the sex-determining locus in A. chinensis. As all Actinidia species are dioecious, we suggest that the sex determining loci of other Actinidia species will be similar to that region defined in our maps. As the extent of the non-recombining region is limited, our result supports the suggestion that the subtelomeric region of an autosome is in the early stages of developing the characteristics of a sex chromosome. The maps provide a reference of genetic information in Actinidia for use in genetic analysis and breeding programs. PMID- 19284546 TI - Early treatment with noninvasive positive pressure ventilation prolongs survival in Amyotrophic Lateral Sclerosis patients with nocturnal respiratory insufficiency. AB - BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease, which rapidly leads to chronic respiratory failure requiring mechanical ventilation. Currently, forced vital capacity (FVC) < 50% is considered as physiologic marker for admitting patients to Noninvasive Positive Pressure Ventilation (NPPV) intervention, although it has been recently shown the median survival of patients with baseline FVC < 75% much shorter than median survival of patients with baseline FVC > 75%, independently by any treatment. AIM: To assess the role of NPPV in improving outcome of ALS, a retrospective analysis was performed to investigate 1 year survival of ALS patients with FVC < 75% and nocturnal respiratory insufficiency, treated with NPPV, compared to a well matched population of ALS patients, who refused or was intolerant to NPPV. METHODS: We investigated seventy-two consecutive ALS patients who underwent pulmonary function test. Forty-four presented a FVC > 75% and served as control group. Twenty-eight patients presented a FVC < 75% and showed, at polysomnography analysis, nocturnal respiratory insufficiency, requiring NPPV; sixteen were treated with NPPV, while twelve refused or were intolerant. RESULTS: Increased survival rate at 1 year in patients with FVC < 75% treated with NPPV, as compared to those who refused or could not tolerate NPPV (p = 0.02), was observed. The median rate of decline in FVC% was slower in NPPV patients than in patients who did not use NPPV (95% CI: 0.72 to 1.85; p < 0.0001). CONCLUSION: This report demonstrates that early treatment with NPPV prolongs survival and reduces decline of FVC% in ALS. PMID- 19284547 TI - Coronary arterial calcification in rheumatoid arthritis: comparison with the Multi-Ethnic Study of Atherosclerosis. AB - INTRODUCTION: Although cardiovascular morbidity and mortality are increased in rheumatoid arthritis, little is known about the burden of subclinical coronary atherosclerosis in these patients. METHODS: Using computed tomography, coronary artery calcification was measured in 195 men and women with rheumatoid arthritis aged 45 to 84 years without clinical cardiovascular disease and compared with 1,073 controls without rheumatoid arthritis enrolled in the Baltimore cohort of the Multi-Ethnic Study of Atherosclerosis. RESULTS: The prevalence of coronary calcification (Agatston score > 0) was significantly higher in men, but not women, with rheumatoid arthritis after adjusting for sociodemographic and cardiovascular risk factors (prevalence ratio = 1.19; P = 0.012). Among participants with prevalent calcification, those with rheumatoid arthritis had adjusted mean Agatston scores 53 units higher than controls (P = 0.002); a difference greater for men than women (P for interaction = 0.017). In all analyses, serum IL-6 attenuated the association between rheumatoid arthritis and coronary calcification, suggesting its role as a potential mediator of enhanced atherosclerosis. Notably, increasing severity of rheumatoid arthritis was associated with a higher prevalence and extent of coronary calcification among both men and women with rheumatoid arthritis, and for all age categories. The largest percentage difference in coronary arterial calcification between rheumatoid arthritis patients and their nonrheumatoid arthritis counterparts was observed in the youngest age category. CONCLUSIONS: Increasing rheumatoid arthritis disease severity was associated with a higher prevalence and greater extent of coronary artery calcification, potentially mediated through an atherogenic effect of chronic systemic inflammation. Gender and age differences in association with coronary calcification suggest that preventive measures should be emphasized in men with rheumatoid arthritis, and considered even in younger rheumatoid arthritis patients with low levels of traditional cardiovascular risk factors. PMID- 19284548 TI - Phenotypes induced by NM causing alpha-skeletal muscle actin mutants in fibroblasts, Sol 8 myoblasts and myotubes. AB - BACKGROUND: Nemaline myopathy is a neuromuscular disorder characterized by the presence of nemaline bodies in patient muscles. 20% of the cases are associated with alpha-skeletal muscle actin mutations. We previously showed that actin mutations can cause four different biochemical phenotypes and that expression of NM associated actin mutants in fibroblasts, myoblasts and myotubes induces a range of cellular defects. FINDINGS: We conducted the same biochemical experiments for twelve new actin mutants associated with nemaline myopathy. We observed folding and polymerization defects. Immunostainings of these and eight other mutants in transfected cells revealed typical cellular defects such as nemaline rods or aggregates, decreased incorporation in F-actin structures, membrane blebbing, the formation of thickened actin fibres and cell membrane blebbing in myotubes. CONCLUSION: Our results confirm that NM associated alpha actin mutations induce a range of defects at the biochemical level as well as in cultured fibroblasts and muscle cells. PMID- 19284549 TI - Gz mediates the long-lasting desensitization of brain CB1 receptors and is essential for cross-tolerance with morphine. AB - BACKGROUND: Although the systemic administration of cannabinoids produces antinociception, their chronic use leads to analgesic tolerance as well as cross tolerance to morphine. These effects are mediated by cannabinoids binding to peripheral, spinal and supraspinal CB1 and CB2 receptors, making it difficult to determine the relevance of each receptor type to these phenomena. However, in the brain, the CB1 receptors (CB1Rs) are expressed at high levels in neurons, whereas the expression of CB2Rs is marginal. Thus, CB1Rs mediate the effects of smoked cannabis and are also implicated in emotional behaviors. We have analyzed the production of supraspinal analgesia and the development of tolerance at CB1Rs by the direct injection of a series of cannabinoids into the brain. The influence of the activation of CB1Rs on supraspinal analgesia evoked by morphine was also evaluated. RESULTS: Intracerebroventricular (icv) administration of cannabinoid receptor agonists, WIN55,212-2, ACEA or methanandamide, generated a dose dependent analgesia. Notably, a single administration of these compounds brought about profound analgesic tolerance that lasted for more than 14 days. This decrease in the effect of cannabinoid receptor agonists was not mediated by depletion of CB1Rs or the loss of regulated G proteins, but, nevertheless, it was accompanied by reduced morphine analgesia. On the other hand, acute morphine administration produced tolerance that lasted only 3 days and did not affect the CB1R. We found that both neural mu-opioid receptors (MORs) and CB1Rs interact with the HINT1-RGSZ module, thereby regulating pertussis toxin-insensitive Gz proteins. In mice with reduced levels of these Gz proteins, the CB1R agonists produced no such desensitization or morphine cross-tolerance. On the other hand, experimental enhancement of Gz signaling enabled an acute icv administration of morphine to produce a long-lasting tolerance at MORs that persisted for more than 2 weeks, and it also impaired the analgesic effects of cannabinoids. CONCLUSION: In the brain, cannabinoids can produce analgesic tolerance that is not associated with the loss of surface CB1Rs or their uncoupling from regulated transduction. Neural specific Gz proteins are essential mediators of the analgesic effects of supraspinal CB1R agonists and morphine. These Gz proteins are also responsible for the long-term analgesic tolerance produced by single doses of these agonists, as well as for the cross-tolerance between CB1Rs and MORs. PMID- 19284550 TI - Uncovering metabolic pathways relevant to phenotypic traits of microbial genomes. AB - Identifying the biochemical basis of microbial phenotypes is a main objective of comparative genomics. Here we present a novel method using multivariate machine learning techniques for comparing automatically derived metabolic reconstructions of sequenced genomes on a large scale. Applying our method to 266 genomes directly led to testable hypotheses such as the link between the potential of microorganisms to cause periodontal disease and their ability to degrade histidine, a link also supported by clinical studies. PMID- 19284551 TI - Monoallelic maternal expression of STAT5A affects embryonic survival in cattle. AB - BACKGROUND: Reproductive disorders and infertility are surprisingly common in the human population as well as in other species. The decrease in fertility is a major cause of cow culling and economic loss in the dairy herd. The conception rate has been declining for the past 30-50 years. Conception rate is the product of fertilization and embryonic survival rates. In a previous study, we have identified associations of several single nucleotide polymorphisms (SNPs) in the signal transducer and activator 5A (STAT5A) with fertilization and survival rates in an in vitro experimental system. The objectives of this study are to fine map the STAT5A region in a search for causative mutations and to investigate the parent of origin expression of this gene. RESULTS: We have performed a total of 5,222 fertilizations and produced a total of 3,696 in vitro fertilized embryos using gametes from 440 cows and eight bulls. A total of 37 SNPs were developed in a 63.4-kb region of genomic sequence that includes STAT5A, STAT3, and upstream and downstream sequences of these genes. SNP153137 (G/C) in exon 8 of STAT5A was associated with a significant variability in embryonic survival and fertilization rate compared to all other examined SNPs. Expression analysis revealed that STAT5A is primarily monoallelically expressed in early embryonic stages but biallelically expressed in later fetal stages. Furthermore, the occurrence of monoallelic maternal expression of STAT5A was significantly higher in blastocysts, while paternal expression was more frequent in degenerative embryos. CONCLUSION: Our results imply that STAT5A affects embryonic survival in a manner influenced by developmental stage and allele parent of origin. PMID- 19284552 TI - Bioinformatic characterizations and prediction of K+ and Na+ ion channels effector toxins. AB - BACKGROUND: K+ and Na+ channel toxins constitute a large set of polypeptides, which interact with their ion channel targets. These polypeptides are classified in two different structural groups. Recently a new structural group called birtoxin-like appeared to contain both types of toxins has been described. We hypothesized that peptides of this group may contain two conserved structural motifs in K+ and/or Na+ channels scorpion toxins, allowing these birtoxin-like peptides to be active on K+ and/or Na+ channels. RESULTS: Four multilevel motifs, overrepresented and specific to each group of K+ and/or Na+ ion channel toxins have been identified, using GIBBS and MEME and based on a training dataset of 79 sequences judged as representative of K+ and Na+ toxins.Unexpectedly birtoxin like peptides appeared to present a new structural motif distinct from those present in K+ and Na+ channels Toxins. This result, supported by previous experimental data, suggests that birtoxin-like peptides may exert their activity on different sites than those targeted by classic K+ or Na+ toxins.Searching, the nr database with these newly identified motifs using MAST, retrieved several sequences (116 with e-value < 1) from various scorpion species (test dataset). The filtering process left 30 new and highly likely ion channel effectors.Phylogenetic analysis was used to classify the newly found sequences. Alternatively, classification tree analysis, using CART algorithm adjusted with the training dataset, using the motifs and their 2D structure as explanatory variables, provided a model for prediction of the activity of the new sequences. CONCLUSION: The phylogenetic results were in perfect agreement with those obtained by the CART algorithm.Our results may be used as criteria for a new classification of scorpion toxins based on functional motifs. PMID- 19284553 TI - New therapeutic approach to Tourette Syndrome in children based on a randomized placebo-controlled double-blind phase IV study of the effectiveness and safety of magnesium and vitamin B6. AB - BACKGROUND: Tourette Syndrome (TS) is a neurological condition presenting chronic motor and phonic tics, and important degree of comorbidity. Considered an uncommon illness, it first becomes apparent during childhood. Current standard treatment only achieves partial control of the condition, and provokes frequent, and sometimes severe, side effects. METHODS AND DESIGN: Main aim: To show that, with respect to placebo treatment, the combination of 0.5 mEq/Kg magnesium and 2 mg/Kg vitamin B6 reduces motor and phonic tics and incapacity in cases of exacerbated TS among children aged 7-14 years, as measured on the Yale Global Tic Severity Scale (YGTSS). Secondary aims: Assess the safety of the treatment. Describe metabolic changes revealed by PET. Measure the impact of the experimental treatment on family life. METHODOLOGY: Randomized, blinded clinical trials. Phase IV study (new proposal for treatment with magnesium and vitamin B6). SCOPE: children in the geographic area of the study group. Recruitment of subjects: to include patients diagnosed with TS, in accordance with DSM-IV criteria (307.23), during a period of exacerbation, and provided none of the exclusion criteria are met. INSTRUMENTATION: clinical data and the YGTSS score will be obtained at the outset of a period of exacerbation (t0). The examinations will be made after 15 (t1), 30 (t2), 60 (t3) and 90 days (t4). PET will be performed at the t0 and t4. We evaluated decrease in the overall score (t0, t1, t2, t3, t4), PET variations, and impact made by the treatment on the patient's life (Psychological General Well-Being Index). DISCUSSION: Few clinical trials have been carried out on children with TS, but they are necessary, as current treatment possibilities are insufficient and often provoke side effects. The difficulty of dealing with an uncommon illness makes designing such a study all the more complicated. The present study seeks to overcome possible methodological problems by implementing a prior, phase II study, in order to calculate the relevant statistical parameters and to determine the safety of the proposed treatment. Providing a collateral treatment with magnesium and vitamin B6 could improve control of the illness and help reduce side effects. This protocol was approved by the Andalusian Government Committee for Clinical Trials (Spain). This study was funded by the Health Department of the Andalusian Regional Government and by the Healthcare Research Fund of the Carlos III Healthcare Institute (Spanish Ministry of Health). TRIAL REGISTRATION: Current Controlled Trials ISRCTN41082378. PMID- 19284554 TI - Reduction of serum IGF-I levels in patients affected with Monoclonal Gammopathies of undetermined significance or Multiple Myeloma. Comparison with bFGF, VEGF and K-ras gene mutation. AB - BACKGROUND: Serum levels of IGF-I in patients affected with multiple myeloma (MM) have been scarcely studied. The present study is aimed to explore this point comparing 55 healthy subjects, 71 monoclonal gammopaties of uncertain significance (MGUS) and 77 overt MM patients. In the same subjects, basic FGF and VEGF, have been detected. All three mediators were analyzed in function of K-ras mutation and melphalan response. Concerning IGF-I, two representative monitoring examples have also been added. METHODS: Cytokine determinations were performed by commercially available ELISA kits, while K12-ras mutation was investigated on genomic DNA isolated from bone marrow cell specimens by RFLP-PCR assay. RESULTS: Significant reductions of IGF-I levels were observed in MGUS and MM as compared with healthy controls. In addition, MM subjects showed significantly decreased serum IGF-I levels than MGUS. Conversely, increasing levels were observed for bFGF and VEGF, molecules significantly correlated. A multivariate analysis corrected for age and gender confirmed the significant difference only for IGF-I values (P = 0.01). K12-ras mutation was significantly associated with malignancy, response to therapy and with significantly increased serum bFGF levels. CONCLUSION: IGF-I reduction in the transition: Controls-->MGUS-->MM and changes observed over time suggest that IGF-I should be furtherly studied in future clinical trials as a possible monitoring marker for MM. PMID- 19284555 TI - delta-Catenin promotes prostate cancer cell growth and progression by altering cell cycle and survival gene profiles. AB - BACKGROUND: delta-Catenin is a unique member of beta-catenin/armadillo domain superfamily proteins and its primary expression is restricted to the brain. However, delta-catenin is upregulated in human prostatic adenocarcinomas, although the effects of delta-catenin overexpression in prostate cancer are unclear. We hypothesized that delta-catenin plays a direct role in prostate cancer progression by altering gene profiles of cell cycle regulation and cell survival. RESULTS: We employed gene transfection and small interfering RNA to demonstrate that increased delta-catenin expression promoted, whereas its knockdown suppressed prostate cancer cell viability. delta-Catenin promoted prostate cancer cell colony formation in soft agar as well as tumor xenograft growth in nude mice. Deletion of either the amino-terminal or carboxyl-terminal sequences outside the armadillo domains abolished the tumor promoting effects of delta-catenin. Quantitative RT2 Profiler PCR Arrays demonstrated gene alterations involved in cell cycle and survival regulation. delta-Catenin overexpression upregulated cyclin D1 and cdc34, increased phosphorylated histone-H3, and promoted the entry of mitosis. In addition, delta-catenin overexpression resulted in increased expression of cell survival genes Bcl-2 and survivin while reducing the cell cycle inhibitor p21Cip1. CONCLUSION: Taken together, our studies suggest that at least one consequence of an increased expression of delta-catenin in human prostate cancer is the alteration of cell cycle and survival gene profiles, thereby promoting tumor progression. PMID- 19284556 TI - Parents' assessment of parent-child interaction interventions--a longitudinal study in 101 families. AB - BACKGROUND: The aim of the study was to describe families with small children who participated in parent-child interaction interventions at four centres in Sweden, and to examine long term and short term changes regarding the parents' experience of parental stress, parental attachment patterns, the parents' mental health and life satisfaction, the parents' social support and the children's problems. METHODS: In this longitudinal study a consecutive sample of 101 families (94 mothers and 54 fathers) with 118 children (median age 3 years) was assessed, using self-reports, at the outset of the treatment (T1), six months later (T2) and 18 months after the beginning of treatment (T3). Analysis of the observed differences was carried out using Wilcoxon's Signed-Rank test and Cohen's d. RESULTS: The results from commencement of treatment showed that the parents had considerable problems in all areas examined. At the outset of treatment (T1) the mothers showed a higher level of problem load than the fathers on almost all scales. In the families where the children's problems have also been measured (children from the age of four) it appeared that they had problems of a nature and degree otherwise found in psychiatric populations. We found a clear general trend towards a positive development from T1 to T2 and this development was also reinforced from T2 to T3. Aggression in the child was one of the most common causes for contact. There were few undesired or unplanned interruptions of the treatment, and the attrition from the study was low. CONCLUSION: This study has shown that it is possible to reach mothers as well as fathers with parenting problems and to create an intervention program with very low dropout levels - which is of special importance for families with small children displaying aggressive behaviour. The parents taking part in this study showed clear improvement trends after six months and this development was reinforced a year later. This study suggests the necessity of clinical development and future research concerning the role of fathers in parent-child interaction interventions. PMID- 19284557 TI - Rheumatoid cachexia is associated with dyslipidemia and low levels of atheroprotective natural antibodies against phosphorylcholine but not with dietary fat in patients with rheumatoid arthritis: a cross-sectional study. AB - INTRODUCTION: Patients with rheumatoid arthritis (RA) have an increased risk for cardiovascular disease (CVD) independent of traditional risk factors. The aim of this study was to analyze the associations between diet, body composition, lipids and atheroprotective natural antibodies against phosphorylcholine (anti-PC) in patients with RA. METHODS: A total of 80 RA patients (76% women), mean age (standard deviation (SD)) 61.4 (12) years and median disease duration of 6 years, were assessed by food frequency questionnaire (FFQ), fatty acid profile in adipose tissue and whole-body dual energy x ray absorptiometry (DXA). Rheumatoid cachexia was defined as fat free mass index below the 25th percentile and fat mass index above the 50th percentile of a reference population. Blood lipids, oxidized low-density lipoprotein (oxLDL) and anti-PC levels were determined. RESULTS: The mean body mass index for the women and men was 25.0 and 27.0, respectively. Central obesity was found in 57% of the women (waist circumference >80 cm) and in 89% of the men (waist circumference >94 cm). In all, 18% of the women and 26% of the men had rheumatoid cachexia. These patients had significantly higher total cholesterol (P < 0.033), LDL (P < 0.029), and trendwise oxLDL (P = 0.056) as well as lower anti-PC IgM (P = 0.040), higher frequency of hypertension (69%) and metabolic syndrome (25%) than those without. The patients reported a high dietary intake of saturated fat, which partly correlated with fatty acid composition in adipose tissue and significantly with disease activity. However, patients with or without cachexia did not differ with respect to dietary fat intake or intake of Mediterranean-like diet. Additionally, patients on a Mediterranean-like diet had high levels of anti-PC (P < 0.001). CONCLUSIONS: About one in five patients with low-active RA displayed rheumatoid cachexia. This condition was associated with high levels of LDL cholesterol, low levels of atheroprotective anti-PC and high frequency of hypertension, which is of interest in the context of CVD in RA. The cachexia could not be related to diet fat intake. However, patients on a Mediterranean-like diet had high anti-PC levels in spite of similar frequency of cachexia. High anti-PC levels may provide some protection against CVD. PMID- 19284558 TI - Candidate autoantigens identified by mass spectrometry in early rheumatoid arthritis are chaperones and citrullinated glycolytic enzymes. AB - INTRODUCTION: The aim of our study was to identify new early rheumatoid arthritis (RA) autoantibodies. METHODS: Sera obtained from 110 early untreated RA patients (<6 months) were analyzed by western blot using HL-60 cell extract, separated on one-dimensional and two-dimensional gel electrophoresis (1-DE, 2-DE). Sera from 50 healthy blood donors and 20 patients with non-RA rheumatisms were used as controls for 1-DE and 2-DE, respectively. The immunoreactive proteins were identified by MALDI-TOF mass spectrometric analysis and the presence of potential sites of citrullination in each of these proteins was evaluated. FT-ICR mass spectrometry was used to verify experimentally the effect of citrullination upon the mass profile observed by MALDI-TOF analysis. RESULTS: The 110 1-DE patterns allowed detection of 10 recurrent immunoreactive bands of 33, 39, 43, 46, 51, 54, 58, 62, 67 and 70 kDa, which were further characterized by 2-DE and proteomic analysis. Six proteins were already described RA antigens: heterogeneous nuclear ribonucleoprotein A2/B1, aldolase, alpha-enolase, calreticulin, 60 kDa heat shock protein (HSP60) and BiP. Phosphoglycerate kinase 1 (PGK1), stress-induced phosphoprotein 1 and the far upstream element-binding proteins (FUSE-BP) 1 and 2 were identified as new antigens. Post-translational protein modifications were analyzed and potentially deiminated peptides were found on aldolase, alpha enolase, PGK1, calreticulin, HSP60 and the FUSE-BPs. We compared the reactivity of RA sera with citrullinated and noncitrullinated alpha-enolase and FUSE-BP linear peptides, and showed that antigenicity of the FUSE-BP peptide was highly dependent on citrullination. Interestingly, the anti-cyclic citrullinated peptide antibody (anti-CCP2) status in RA serum at inclusion was not correlated to the reactivity directed against FUSE-BP citrullinated peptide. CONCLUSIONS: Two categories of antigens, enzymes of the glycolytic family and molecular chaperones are also targeted by the early untreated RA autoantibody response. For some of them, and notably the FUSE-BPs, citrullination is involved in the immunological tolerance breakdown observed earlier in RA patients. Autoantibodies recognizing a citrullinated peptide from FUSE-BP may enhance the sensibility for RA of the currently available anti-CCP2 test. PMID- 19284559 TI - The relative ages of ectomycorrhizal mushrooms and their plant hosts estimated using Bayesian relaxed molecular clock analyses. AB - BACKGROUND: Ectomycorrhizae (ECM) are symbioses formed by polyphyletic assemblages of fungi (mostly Agaricomycetes) and plants (mostly Pinaceae and angiosperms in the rosid clade). Efforts to reconstruct the evolution of the ECM habit in Agaricomycetes have yielded vastly different results, ranging from scenarios with many relatively recent origins of the symbiosis and no reversals to the free-living condition; a single ancient origin of ECM and many subsequent transitions to the free-living condition; or multiple gains and losses of the association. To test the plausibility of these scenarios, we performed Bayesian relaxed molecular clock analyses including fungi, plants, and other eukaryotes, based on the principle that a symbiosis cannot evolve prior to the origin of both partners. As we were primarily interested in the relative ages of the plants and fungi, we did not attempt to calibrate the molecular clock using the very limited fossil record of Agaricomycetes. RESULTS: Topologically constrained and unconstrained analyses suggest that the root node of the Agaricomycetes is much older than either the rosids or Pinaceae. The Agaricomycetidae, a large clade containing the Agaricales and Boletales (collectively representing 70% of Agaricomycetes), is also significantly older than the rosids. The relative age of Agaricomycetidae and Pinaceae, however, is sensitive to tree topology, and the inclusion or exclusion of the gnetophyte Welwitschia mirabilis. CONCLUSION: The ancestor of the Agaricomycetes could not have been an ECM species because it existed long before any of its potential hosts. Within more derived clades of Agaricomycetes, there have been at least eight independent origins of ECM associations involving angiosperms, and at least six to eight origins of associations with gymnosperms. The first ECM symbioses may have involved Pinaceae, which are older than rosids, but several major clades of Agaricomycetes, such as the Boletales and Russulales, are young enough to have been plesiomorphically associated with either rosids or Pinaceae, suggesting that some contemporary ECM partnerships could be of very ancient origin. PMID- 19284560 TI - Metabolism during anaesthesia and recovery in colic and healthy horses: a microdialysis study. AB - BACKGROUND: Muscle metabolism in horses has been studied mainly by analysis of substances in blood or plasma and muscle biopsy specimens. By using microdialysis, real-time monitoring of the metabolic events in local tissue with a minimum of trauma is possible. There is limited information about muscle metabolism in the early recovery period after anaesthesia in horses and especially in the colic horse. The aims were to evaluate the microdialysis technique as a complement to plasma analysis and to study the concentration changes in lactate, pyruvate, glucose, glycerol, and urea during anaesthesia and in the recovery period in colic horses undergoing abdominal surgery and in healthy horses not subjected to surgery. METHODS: Ten healthy university-owned horses given anaesthesia alone and ten client-owned colic horses subjected to emergency abdominal surgery were anaesthetised for a mean (range) of 230 min (193 273) and 208 min (145-300) respectively. Venous blood samples were taken before anaesthesia. Venous blood sampling and microdialysis in the gluteal muscle were performed during anaesthesia and until 24 h after anaesthesia. Temporal changes and differences between groups were analysed with an ANOVA for repeated measures followed by Tukey Post Hoc test or Planned Comparisons. RESULTS: Lactate, glucose and urea, in both dialysate and plasma, were higher in the colic horses than in the healthy horses for several hours after recovery to standing. In the colic horses, lactate, glucose, and urea in dialysate, and lactate in plasma increased during the attempts to stand. The lactate-to-pyruvate ratio was initially high in sampled colic horses but decreased over time. In the colic horses, dialysate glycerol concentrations varied considerably whereas in the healthy horses, dialysate glycerol was elevated during anaesthesia but decreased after standing. In both groups, lactate concentration was higher in dialysate than in plasma. The correspondence between dialysate and plasma concentrations of glucose, urea and glycerol varied. CONCLUSION: Microdialysis proved to be suitable in the clinical setting for monitoring of the metabolic events during anaesthesia and recovery. It was possible with this technique to show greater muscle metabolic alterations in the colic horses compared to the healthy horses in response to regaining the standing position. PMID- 19284561 TI - Performance of the Genotype MTBDRPlus assay in the diagnosis of tuberculosis and drug resistance in Samara, Russian Federation. AB - BACKGROUND: Russia is a high tuberculosis (TB) burden country with a high prevalence of multidrug resistant tuberculosis (MDRTB). Molecular assays for detection of MDRTB on clinical specimens are not widely available in Russia. RESULTS: We performed an evaluation of the GenoType MTBDRplus assay (HAIN Lifescience GmbH, Germany) on a total of 168 sputum specimens from individual patients at a public health laboratory in Central Russia, as a model of a middle income site in a region with high levels of drug resistance. Phenotypic drug resistance tests (DST) were performed on cultures derived from the same sputum specimens using the BACTEC 960 liquid media system.Interpretable GenoType MTBDRplus results were obtained for 154(91.7%) specimens with readability rates significantly higher in sputum specimens graded 2+ and 3+ compared to 1+ (RR = 1.17 95%CI 1.04-1.32). The sensitivity and specificity of the assay for the detection of rifampicin (RIF) and isoniazid (INH) resistance and MDR was 96.2%, 97.4%, 97.1% and 90.7%, 83.3%, 88.9% respectively. Mutations in codon 531 of the rpoB gene and codon 315 of the katG gene dominated in RIF and INH resistant strains respectively. Disagreements between phenotypical and molecular tests results (12 samples) could be explained by the presence of rare mutations in strains circulating in Russia and simultaneous presence of resistant and sensitive bacilli in sputum specimens (heteroresistance). CONCLUSION: High sensitivity, short turnaround times and the potential for screening large numbers of specimens rapidly, make the GenoType MTBDRplus assay suitable as a first-line screening assay for drug resistant TB. PMID- 19284562 TI - School-based health centers: A four year experience, with a focus on reducing student exclusion rates. AB - We describe a four year collaborative experience with an on-site, community school-based health center that is staffed by the Vallejo City Unified School District and supervised by the pediatric faculty of the Touro University College of Osteopathic Medicine, with particular attention to first grade student exclusion rates.Patient demographics (including payer source), first grade enrollment statistics, and first grade exclusion rates were analyzed using school district enrollment and exclusion data, billing data, and Child Health Disability Program data.An ethnically diverse patient population is described, with the payer source in 99% of patients being the State of California Child Health Disability Program or no insurance source. Ninety-one percent of office visits were for well child care and immunizations. First grade student exclusion rates for failure to meet the state-mandated physical examination requirement fell 74% over the first four years of the school-based health center's operation.In summary, our school-based health center serves a patient population that is primarily uninsured. Reduction in first grade student exclusion rates enhances student education and reduces the loss of attendance-based state matching funds. Additionally, our school-based health center has been well accepted by the local community. PMID- 19284563 TI - Small molecule activators of SIRT1 replicate signaling pathways triggered by calorie restriction in vivo. AB - BACKGROUND: Calorie restriction (CR) produces a number of health benefits and ameliorates diseases of aging such as type 2 diabetes. The components of the pathways downstream of CR may provide intervention points for developing therapeutics for treating diseases of aging. The NAD+-dependent protein deacetylase SIRT1 has been implicated as one of the key downstream regulators of CR in yeast, rodents, and humans. Small molecule activators of SIRT1 have been identified that exhibit efficacy in animal models of diseases typically associated with aging including type 2 diabetes. To identify molecular processes induced in the liver of mice treated with two structurally distinct SIRT1 activators, SIRT501 (formulated resveratrol) and SRT1720, for three days, we utilized a systems biology approach and applied Causal Network Modeling (CNM) on gene expression data to elucidate downstream effects of SIRT1 activation. RESULTS: Here we demonstrate that SIRT1 activators recapitulate many of the molecular events downstream of CR in vivo, such as enhancing mitochondrial biogenesis, improving metabolic signaling pathways, and blunting pro-inflammatory pathways in mice fed a high fat, high calorie diet. CONCLUSION: CNM of gene expression data from mice treated with SRT501 or SRT1720 in combination with supporting in vitro and in vivo data demonstrates that SRT501 and SRT1720 produce a signaling profile that mirrors CR, improves glucose and insulin homeostasis, and acts via SIRT1 activation in vivo. Taken together these results are encouraging regarding the use of small molecule activators of SIRT1 for therapeutic intervention into type 2 diabetes, a strategy which is currently being investigated in multiple clinical trials. PMID- 19284564 TI - Operating theatre related syncope in medical students: a cross sectional study. AB - BACKGROUND: Observing surgical procedures is a beneficial educational experience for medical students during their surgical placements. Anecdotal evidence suggests that operating theatre related syncope may have detrimental effects on students' views of this. Our study examines the frequency and causes of such syncope, together with effects on career intentions, and practical steps to avoid its occurrence. METHODS: All penultimate and final year students at a large UK medical school were surveyed using the University IT system supplemented by personal approach. A 20-item anonymous questionnaire was distributed and results were analysed using the Statistical Package for Social Sciences, version 15.0 (Chicago, Illinois, USA). RESULTS: Of the 630 clinical students surveyed, 77 responded with details of at least one near or actual operating theatre syncope (12%). A statistically significant gender difference existed for syncopal/near syncopal episodes (male 12%; female 88%), p < 0.05. Twenty-two percent of those affected were graduate entry medical course students with the remaining 78% undergraduate. Mean age was 23-years (range 20 - 45). Of the 77 reactors, 44 (57%) reported an intention to pursue a surgical career. Of this group, 7 (9%) reported being discouraged by syncopal episodes in the operating theatre. The most prevalent contributory factors were reported as hot temperature (n = 61, 79%), prolonged standing (n = 56, 73%), wearing a surgical mask (n = 36, 47%) and the smell of diathermy (n = 18, 23%). The most frequently reported measures that students found helpful in reducing the occurrence of syncopal episodes were eating and drinking prior to attending theatre (n = 47, 61%), and moving their legs whilst standing (n = 14, 18%). CONCLUSION: Our study shows that operating theatre related syncope among medical students is common, and we establish useful risk factors and practical steps that have been used to prevent its occurrence. Our study also highlights the detrimental effect of this on the career intentions of medical students interested in surgery. Based on these findings, we recommend that dedicated time should be set aside in surgical teaching to address this issue prior to students attending the operating theatre. PMID- 19284565 TI - Determinants of subject visit participation in a prospective cohort study of HTLV infection. AB - BACKGROUND: Understanding participation in a prospective study is crucial to maintaining and improving retention rates. In 1990-92, following attempted blood donation at five blood centers, we enrolled 155 HTLV-I, 387 HTLV-II and 799 HTLV seronegative persons in a long-term prospective cohort. METHODS: Health questionnaires and physical exams were administered at enrollment and 2-year intervals through 2004. To examine factors influencing attendance at study visits of the cohort participants we calculated odds ratios (ORs) with generalized estimated equations (GEE) to analyze fixed and time-varying predictors of study visit participation. RESULTS: There were significant independent associations between better visit attendance and female gender (OR = 1.31), graduate education (OR = 1.86) and income > $75,000 (OR = 2.68). Participants at two centers (OR = 0.47, 0.67) and of Black race/ethnicity (OR = 0.61) were less likely to continue. Higher subject reimbursement for interview was associated with better visit attendance (OR = 1.84 for $25 vs. $10). None of the health related variables (HTLV status, perceived health status and referral to specialty diagnostic exam for potential adverse health outcomes) significantly affected participation after controlling for demographic variables. CONCLUSION: Increasing and maintaining participation by minority and lower socioeconomic status participants is an ongoing challenge in the study of chronic disease outcomes. Future studies should include methods to evaluate attrition and retention, in addition to primary study outcomes, including qualitative analysis of reasons for participation or withdrawal. PMID- 19284566 TI - Time course analysis of RNA stability in human placenta. AB - BACKGROUND: Evaluation of RNA quality is essential for gene expression analysis, as the presence of degraded samples may influence the interpretation of expression levels. Particularly, qRT-PCR data can be affected by RNA integrity and stability. To explore systematically how RNA quality affects qRT-PCR assay performance, a set of human placenta RNA samples was generated by two protocols handlings of fresh tissue over a progressive time course of 4 days. Protocol A consists of a direct transfer of tissue into RNA-stabilizing solution (RNAlater) solution. Protocol B uses a dissection of placenta villosities before bio banking. We tested and compared RNA yields, total RNA integrity, mRNA integrity and stability in these two protocols according to the duration of storage. RESULTS: A long time tissue storage had little effect on the total RNA and mRNA integrity but induced changes in the transcript levels of stress-responsive genes as TNF-alpha or COX2 after 48 h. The loss of the RNA integrity was higher in the placental tissues that underwent a dissection before RNA processing by comparison with those transferred directly into RNA later solution. That loss is moderate, with average RIN (RNA Integration Numbers) range values of 4.5-6.05, in comparison with values of 6.44-7.22 in samples directly transferred to RNAlater (protocol A). Among the house keeping genes tested, the B2M is the most stable. CONCLUSION: This study shows that placental samples can be stored at + 4 degrees C up to 48 h before RNA extraction without altering RNA quality. Rapid tissue handling without dissection and using RNA-stabilizing solution (RNAlater) is a prerequisite to obtain suitable RNA integrity and stability. PMID- 19284567 TI - Anti-inflammatory effects of the gorgonian Pseudopterogorgia elisabethae collected at the Islands of Providencia and San Andres (SW Caribbean). AB - BACKGROUND: We are reporting for the first time the in vivo anti-inflammatory activity of extracts and fractions, and in vitro anti-inflammatory activity of pure compounds, all isolated from Pseudopterogorgia elisabethae collected at the Providencia (chemotype 1) and San Andres (chemotype 2) Islands (SW Caribbean). METHODS: Extracts from P. elisabethae were fractionated on silica gel to yield fractions: F-1 (pseudopterosins PsQ, PsS and PsU) and F-2 (amphilectosins A and B, PsG, PsK, PsP and PsT and seco-pseudopterosins seco-PsJ and seco-PsK) from chemotype 1, and F-3 (elisabethatrienol, 10-acetoxy-9-hydroxy- and 9-acetoxy-10 hydroxy-amphilecta-8,10,12,14-tetraenes (interconverting mixture) and amphilecta 8(13),11,14-triene-9,10-dione) from chemotype 2. By using preparative RP-HPLC and spectroscopic means, we obtained the pure PsG, PsK, PsP, PsQ, PsS, PsT, PsU, seco PsK and the interconverting mixture of non-glycosylated diterpenes (IMNGD). The anti-inflammatory properties of extracts and fractions were evaluated using in vivo model "12-O-tetradecanoyl-phorbol-acetate (TPA)-induced mouse ear oedema". The activities of pure compounds and of the IMNGD were evaluated using in vitro assays myeloperoxidase (MPO) release (by human polymorphonuclear neutrophils (PMNs)), nitric oxide release (by J-774 cells) and scavenger activity on NO. RESULTS: In the in vivo anti-inflammatory assay, extracts and F-3 showed low inhibition levels of inflammation compared to indomethacin, F-1 and F-2. Additionally, we evaluated the MPO release to the inflammation site, and found a marked inhibition of MPO levels by all extracts and fractions, even superior to the inhibition shown by indomethacin.Furthermore, in the MPO in vitro assay, IMNGD, PsQ, PsS, PsT and PsU, exhibited higher inhibition levels compared to dexamethasone and indomethacin. In the NO release in vitro, IMNGD, PsP and PsT were the most potent treatments. Finally, because the PsG, PsP and seco-PsK did not exhibit any NO scavenger activity, they should inhibit the inducible Nitric Oxide Synthase (iNOS) or other routes that influence this enzyme. Alternatively, PsQ, PsS, and PsU did show scavenger activity. CONCLUSION: All results presented contribute to demonstrate that the compounds isolated in this work from P. elisabethae are promising molecules with an interesting anti-inflammatory activity profile. Additionally, the results obtained could provide preliminary insights towards their structure-activity relationship. PMID- 19284568 TI - Characterization of STAT3 activation and expression in canine and human osteosarcoma. AB - BACKGROUND: Dysregulation of signal transducer and activator of transcription 3 (STAT3) has been implicated as a key participant in tumor cell survival, proliferation, and metastasis and is often correlated with a more malignant tumor phenotype. STAT3 phosphorylation has been demonstrated in a subset of human osteosarcoma (OSA) tissues and cell lines. OSA in the canine population is known to exhibit a similar clinical behavior and molecular biology when compared to its human counterpart, and is often used as a model for preclinical testing of novel therapeutics. The purpose of this study was to investigate the potential role of STAT3 in canine and human OSA, and to evaluate the biologic activity of a novel small molecule STAT3 inhibitor. METHODS: To examine STAT3 and Src expression in OSA, we performed Western blotting and RT-PCR. OSA cells were treated with either STAT3 siRNA or small molecule Src (SU6656) or STAT3 (LLL3) inhibitors and cell proliferation (CyQUANT), caspase 3/7 activity (ELISA), apoptosis (Western blotting for PARP cleavage) and/or viability (Wst-1) were determined. Additionally, STAT3 DNA binding after treatment was determined using EMSA. Expression of STAT3 targets after treatment was demonstrated with Western blotting, RT-PCR, or gel zymography. RESULTS: Our data demonstrate that constitutive activation of STAT3 is present in a subset of canine OSA tumors and human and canine cell lines, but not normal canine osteoblasts. In both canine and human OSA cell lines, downregulation of STAT3 activity through inhibition of upstream Src family kinases using SU6656, inhibition of STAT3 DNA binding and transcriptional activities using LLL3, or modulation of STAT3 expression using siRNA, all resulted in decreased cell proliferation and viability, ultimately inducing caspase-3/7 mediated apoptosis in treated cells. Furthermore, inhibition of either Src or STAT3 activity downregulated the expression of survivin, VEGF, and MMP2, all known transcriptional targets of STAT3. CONCLUSION: These data suggest that STAT3 activation contributes to the survival and proliferation of human and canine OSA cells, thereby providing a potentially promising target for therapeutic intervention. Future investigational trials of LLL3 in dogs with spontaneous OSA will help to more accurately define the role of STAT3 in the clinical setting. PMID- 19284569 TI - HIV-related risk behaviours and the correlates among rickshaw pullers of Kamrangirchar, Dhaka, Bangladesh: a cross-sectional study using probability sampling. AB - BACKGROUND: National HIV serological and behavioural surveillance of Bangladesh repeatedly demonstrated a very high proportion of rickshaw pullers in Dhaka city, having sex with female sex workers (FSWs) and using illicit substances. However, no study has been conducted to identify the correlates of having sex with FSWs among this population. This study aimed to describe behavioural profile of rickshaw pullers in Dhaka city using probability samples and to identify the correlates for having sex with FSWs in order to focus HIV prevention intervention. METHODS: Six hundred rickshaw pullers were randomly selected from rickshaw garages in the Kamrangirchar area, the single largest slum cluster of Dhaka, Bangladesh, during March-April 2008 using the Proportion Probability to Size method. Participants were interviewed, with a response rate of 99.2% (n = 595), using a structured questionnaire and asked about illicit substance use, sexual behaviour and risk perception for HIV and sexually transmitted diseases. Independent predictors of having sex with FSWs were analysed by multivariate analysis. A qualitative study was subsequently conducted with 30 rickshaw pullers to supplement the findings of the initial survey. RESULTS: The proportion of survey respondents who had sex with FSWs and those who used illicit substances in the previous 12 months period were 7.9% and 24.9%, respectively, much lower than the results achieved in the 2003-04 behavioural surveillance (72.8% and 89.9%, respectively). Multivariate analysis revealed the characteristics of younger age, being never married, living alone with family remaining in other districts and using illicit substances in the previous 12 months were significantly associated with having sex with FSWs. CONCLUSION: HIV-related risk behaviour of our study population of the rickshaw pullers was lower than what has been suggested by the results of behavioural surveillance. While this discrepancy should be addressed in further studies, our study emphasizes the importance of focused HIV prevention programs for rickshaw pullers as high-risk behaviour is displayed at an unacceptable level and concentrated in identifiable sub-populations. PMID- 19284570 TI - Cost effectiveness of antimicrobial catheters in the intensive care unit: addressing uncertainty in the decision. AB - INTRODUCTION: Some types of antimicrobial-coated central venous catheters (A-CVC) have been shown to be cost effective in preventing catheter-related bloodstream infection (CR-BSI). However, not all types have been evaluated, and there are concerns over the quality and usefulness of these earlier studies. There is uncertainty amongst clinicians over which, if any, A-CVCs to use. We re-evaluated the cost effectiveness of all commercially available A-CVCs for prevention of CR BSI in adult intensive care unit (ICU) patients. METHODS: We used a Markov decision model to compare the cost effectiveness of A-CVCs relative to uncoated catheters. Four catheter types were evaluated: minocycline and rifampicin (MR) coated catheters, silver, platinum and carbon (SPC)-impregnated catheters, and two chlorhexidine and silver sulfadiazine-coated catheters; one coated on the external surface (CH/SSD (ext)) and the other coated on both surfaces (CH/SSD (int/ext)). The incremental cost per quality-adjusted life year gained and the expected net monetary benefits were estimated for each. Uncertainty arising from data estimates, data quality and heterogeneity was explored in sensitivity analyses. RESULTS: The baseline analysis, with no consideration of uncertainty, indicated all four types of A-CVC were cost-saving relative to uncoated catheters. MR-coated catheters prevented 15 infections per 1,000 catheters and generated the greatest health benefits, 1.6 quality-adjusted life years, and cost savings (AUD $130,289). After considering uncertainty in the current evidence, the MR-coated catheters returned the highest incremental monetary net benefits of AUD $948 per catheter; however there was a 62% probability of error in this conclusion. Although the MR-coated catheters had the highest monetary net benefits across multiple scenarios, the decision was always associated with high uncertainty. CONCLUSIONS: Current evidence suggests that the cost effectiveness of using A-CVCs within the ICU is highly uncertain. Policies to prevent CR-BSI amongst ICU patients should consider the cost effectiveness of competing interventions in the light of this uncertainty. Decision makers would do well to consider the current gaps in knowledge and the complexity of producing good quality evidence in this area. PMID- 19284571 TI - Quarantine for pandemic influenza control at the borders of small island nations. AB - BACKGROUND: Although border quarantine is included in many influenza pandemic plans, detailed guidelines have yet to be formulated, including considerations for the optimal quarantine length. Motivated by the situation of small island nations, which will probably experience the introduction of pandemic influenza via just one airport, we examined the potential effectiveness of quarantine as a border control measure. METHODS: Analysing the detailed epidemiologic characteristics of influenza, the effectiveness of quarantine at the borders of islands was modelled as the relative reduction of the risk of releasing infectious individuals into the community, explicitly accounting for the presence of asymptomatic infected individuals. The potential benefit of adding the use of rapid diagnostic testing to the quarantine process was also considered. RESULTS: We predict that 95% and 99% effectiveness in preventing the release of infectious individuals into the community could be achieved with quarantine periods of longer than 4.7 and 8.6 days, respectively. If rapid diagnostic testing is combined with quarantine, the lengths of quarantine to achieve 95% and 99% effectiveness could be shortened to 2.6 and 5.7 days, respectively. Sensitivity analysis revealed that quarantine alone for 8.7 days or quarantine for 5.7 days combined with using rapid diagnostic testing could prevent secondary transmissions caused by the released infectious individuals for a plausible range of prevalence at the source country (up to 10%) and for a modest number of incoming travellers (up to 8000 individuals). CONCLUSION: Quarantine at the borders of island nations could contribute substantially to preventing the arrival of pandemic influenza (or at least delaying the arrival date). For small island nations we recommend consideration of quarantine alone for 9 days or quarantine for 6 days combined with using rapid diagnostic testing (if available). PMID- 19284572 TI - Topological and organizational properties of the products of house-keeping and tissue-specific genes in protein-protein interaction networks. AB - BACKGROUND: Human cells of various tissue types differ greatly in morphology despite having the same set of genetic information. Some genes are expressed in all cell types to perform house-keeping functions, while some are selectively expressed to perform tissue-specific functions. In this study, we wished to elucidate how proteins encoded by human house-keeping genes and tissue-specific genes are organized in human protein-protein interaction networks. We constructed protein-protein interaction networks for different tissue types using two gene expression datasets and one protein-protein interaction database. We then calculated three network indices of topological importance, the degree, closeness, and betweenness centralities, to measure the network position of proteins encoded by house-keeping and tissue-specific genes, and quantified their local connectivity structure. RESULTS: Compared to a random selection of proteins, house-keeping gene-encoded proteins tended to have a greater number of directly interacting neighbors and occupy network positions in several shortest paths of interaction between protein pairs, whereas tissue-specific gene-encoded proteins did not. In addition, house-keeping gene-encoded proteins tended to connect with other house-keeping gene-encoded proteins in all tissue types, whereas tissue-specific gene-encoded proteins also tended to connect with other tissue-specific gene-encoded proteins, but only in approximately half of the tissue types examined. CONCLUSION: Our analysis showed that house-keeping gene encoded proteins tend to occupy important network positions, while those encoded by tissue-specific genes do not. The biological implications of our findings were discussed and we proposed a hypothesis regarding how cells organize their protein tools in protein-protein interaction networks. Our results led us to speculate that house-keeping gene-encoded proteins might form a core in human protein protein interaction networks, while clusters of tissue-specific gene-encoded proteins are attached to the core at more peripheral positions of the networks. PMID- 19284573 TI - Studies on the binding affinity of anticancer drug mitoxantrone to chromatin, DNA and histone proteins. AB - Mitoxantrone is a potent antitumor drug, widely used in the treatment of various cancers. In the present study, we have investigated and compared the affinity of anticancer drug, mitoxantrone, to EDTA-soluble chromatin (SE-chromatin), DNA and histones employing UV/Vis, fluorescence, CD spectroscopy, gel electrophoresis and equilibrium dialysis techniques. The results showed that the interaction of mitoxantrone with SE-chromatin proceeds into compaction/aggregation as revealed by reduction in the absorbencies at 608 and 260 nm (hypochromicity) and disappearance of both histones and DNA on the gels. Mitoxantrone interacts strongly with histone proteins in solution making structural changes in the molecule as shown by CD and fluorescence analysis. The binding isotherms demonstrate a positive cooperative binding pattern for the chromatin- mitoxantrone interaction. It is suggested higher binding affinity of mitoxantrone to chromatin compared to DNA implying that the histone proteins may play an important role in the chromatin- mitoxantrone interaction process. PMID- 19284574 TI - A role for CD81 on the late steps of HIV-1 replication in a chronically infected T cell line. AB - BACKGROUND: HIV-1 uses cellular co-factors for virion formation and release. The virus is able to incorporate into the viral particles host cellular proteins, such as tetraspanins which could serve to facilitate HIV-1 egress. Here, we investigated the implication of several tetraspanins on HIV-1 formation and release in chronically infected T-lymphoblastic cells, a model that permits the study of the late steps of HIV-1 replication. RESULTS: Our data revealed that HIV 1 Gag and Env structural proteins co-localized with tetraspanins in the form of clusters. Co-immunoprecipitation experiments showed that Gag proteins interact, directly or indirectly, with CD81, and less with CD82, in tetraspanin-enriched microdomains composed of CD81/CD82/CD63. In addition, when HIV-1 producing cells were treated with anti-CD81 antibodies, or upon CD81 silencing by RNA interference, HIV-1 release was significantly impaired, and its infectivity was modulated. Finally, CD81 downregulation resulted in Gag redistribution at the cell surface. CONCLUSION: Our findings not only extend the notion that HIV-1 assembly can occur on tetraspanin-enriched microdomains in T cells, but also highlight a critical role for the tetraspanin CD81 on the late steps of HIV replication. PMID- 19284575 TI - Lack of utility of risk score and gynecological examination for screening for sexually transmitted infections in sexually active adolescents. AB - BACKGROUND: Sexually transmitted infections constitute the main health risk among adolescents. In developing countries the diagnosis and treatment of cervical infections is based on the syndromic approach. In this study we estimated the prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae among female adolescents from a Health Sector of the city of Goiania, Brazil, and validated cervicitis diagnosis using World Health Organization/Ministry of Health risk score and gynecological examination. METHODS: A cross-sectional community-based sample of 914 15- to 19-year-old female teenagers was randomly selected and referred to the local Family Health Program. Of these, 472 (51.6%) were sexually active and gynecological examinations were carried out for 427. Endocervical samples were collected to perform the polymerase chain reaction for C. trachomatis and N. gonorrhoeae. Performance of risk score, the presence of mucopurulent discharge, friability, ectopia and pain during cervical maneuver were compared with the presence of C. trachomatis or N. gonorrhoeae or both. RESULTS: The prevalence of C. trachomatis and N. gonorrhoeae was 14.5% and 2.1%, respectively. The risk score had a specificity of 31.9% (95% confidence interval, 21.2 to 44.2) and a positive predictive value of 20.8% (95% confidence interval, 13.5 to 29.7). Friability was the component of the gynecological examination that presented the best performance with a sensitivity of 43.5%, specificity of 81.0%, and 30.6% of positive predictive value. CONCLUSION: The prevalence of infection by C. trachomatis and N. gonorrhoeae was high among these sexually active adolescents. The syndromic approach is clearly inadequate for screening and treating these infections in this population. Therefore, the implantation of other strategies to control these infections among adolescents is urgently required. PMID- 19284576 TI - Validation study of villous atrophy and small intestinal inflammation in Swedish biopsy registers. AB - BACKGROUND: Small intestinal biopsy with villous atrophy (VA) is the gold standard for the diagnosis of celiac disease (CD). We validated VA (Marsh 3) and small intestinal inflammation without VA (Marsh 1+2) in Swedish regional biopsy registers. METHODS: All pathology departments in Sweden (n = 28) were searched to identify individuals with VA or duodenal/jejunal inflammation. The validation consisted of blinded examination of biopsy samples, manual review of biopsy reports, web surveys, and patient chart reviews of 121 individuals with VA and 39 with inflammation. RESULTS: We identified 29,148 individuals with VA and 13,446 individuals with inflammation. In a blinded examination, Swedish pathologists correctly classified 90% of biopsies with VA. Manual screening of 1,534 biopsy reports (performed by co-author JFL and a research assistant) found that comorbidity other than CD was rare. IBD was the most common comorbidity and occurred in 0.3% of biopsies with VA (1.6% in inflammation). Among 114 patients with VA and available data, 108 (95%) had a clinical diagnosis of CD. 79% of the validated individuals with VA and 64% of those with inflammation had documented gastrointestinal symptoms prior to biopsy. 88% of the validated individuals with VA had positive CD serology before their first biopsy. 172/180 (96%) of Swedish gastroenterologists and 68/68 (100%) of pediatricians perform a small intestinal biopsy in at least 9 out of 10 individuals prior to diagnosis of CD. CONCLUSION: Regional biopsy data are feasible to identify individuals with CD and small intestinal inflammation. The specificity of CD is high in villous atrophy. PMID- 19284577 TI - Serum levels of soluble receptor for advanced glycation end products and of S100 proteins are associated with inflammatory, autoantibody, and classical risk markers of joint and vascular damage in rheumatoid arthritis. AB - INTRODUCTION: The receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin superfamily of cell surface receptor molecules. High concentrations of three of its putative proinflammatory ligands, S100A8/A9 complex (calprotectin), S100A8, and S100A12, are found in rheumatoid arthritis (RA) serum and synovial fluid. In contrast, soluble RAGE (sRAGE) may prevent proinflammatory effects by acting as a decoy. This study evaluated the serum levels of S100A9, S100A8, S100A12 and sRAGE in RA patients, to determine their relationship to inflammation and joint and vascular damage. METHODS: Serum sRAGE, S100A9, S100A8 and S100A12 levels from 138 patients with established RA and 44 healthy controls were measured by ELISA and compared by unpaired t test. In RA patients, associations with disease activity and severity variables were analyzed by simple and multiple linear regressions. RESULTS: Serum S100A9, S100A8 and S100A12 levels were correlated in RA patients. S100A9 levels were associated with body mass index (BMI), and with serum levels of S100A8 and S100A12. S100A8 levels were associated with serum levels of S100A9, presence of anti-citrullinated peptide antibodies (ACPA), and rheumatoid factor (RF). S100A12 levels were associated with presence of ACPA, history of diabetes, and serum S100A9 levels. sRAGE levels were negatively associated with serum levels of C-reactive protein (CRP) and high-density lipoprotein (HDL), history of vasculitis, and the presence of the RAGE 82Ser polymorphism. CONCLUSIONS: sRAGE and S100 proteins were associated not just with RA inflammation and autoantibody production, but also with classical vascular risk factors for end-organ damage. Consistent with its role as a RAGE decoy molecule, sRAGE had the opposite effects to S100 proteins in that S100 proteins were associated with autoantibodies and vascular risk, whereas sRAGE was associated with protection against joint and vascular damage. These data suggest that RAGE activity influences co-development of joint and vascular disease in rheumatoid arthritis patients. PMID- 19284578 TI - A novel method for producing target cells and assessing cytotoxic T lymphocyte activity in outbred hosts. AB - BACKGROUND: Cytotoxic T lymphocytes play a crucial role in the immunological control of microbial infections and in the design of vaccines and immunotherapies. Measurement of cytotoxic T lymphocyte activity requires that the test antigen is presented by target cells having the same or compatible class I major histocompatibility complex antigens as the effector cells. Conventional assays use target cells labeled with 51chromium and infer cytotoxic T lymphocyte activity by measuring the isotope released by the target cells lysed following incubation with antigen-specific cytotoxic T lymphocytes. This assay is sensitive but needs manipulation and disposal of hazardous radioactive reagents and provides a bulk estimate of the reporter released, which may be influenced by spontaneous release of the label and other poorly controllable variables. Here we describe a novel method for producing target in outbred hosts and assessing cytotoxic T lymphocyte activity by flow cytometry. RESULTS: The method consists of culturing skin fibroblasts, immortalizing them with a replication defective clone of simian virus 40, and finally transducing them with a bicistronic vector encoding the target antigen and the reporter green fluorescent protein. When used in a flow cytometry-based assay, the target cells obtained with this method proved valuable for assessing the viral envelope protein specific cytotoxic T lymphocyte activity in domestic cats acutely or chronically infected with feline immunodeficiency virus, a lentivirus similar to human immunodeficiency virus and used as animal model for AIDS studies. CONCLUSION: Given the versatility of the bicistronic vector used, its ability to deliver multiple and large transgenes in target cells, and its extremely wide cell specificity when pseudotyped with the vesicular stomatitis virus envelope protein, the method is potentially exploitable in many animal species. PMID- 19284579 TI - Evolution from a respiratory ancestor to fill syntrophic and fermentative niches: comparative fenomics of six Geobacteraceae species. AB - BACKGROUND: The anaerobic degradation of organic matter in natural environments, and the biotechnical use of anaerobes in energy production and remediation of subsurface environments, both require the cooperative activity of a diversity of microorganisms in different metabolic niches. The Geobacteraceae family contains members with three important anaerobic metabolisms: fermentation, syntrophic degradation of fermentation intermediates, and anaerobic respiration. RESULTS: In order to learn more about the evolution of anaerobic microbial communities, the genome sequences of six Geobacteraceae species were analyzed. The results indicate that the last common Geobacteraceae ancestor contained sufficient genes for anaerobic respiration, completely oxidizing organic compounds with the reduction of external electron acceptors, features that are still retained in modern Geobacter and Desulfuromonas species. Evolution of specialization for fermentative growth arose twice, via distinct lateral gene transfer events, in Pelobacter carbinolicus and Pelobacter propionicus. Furthermore, P. carbinolicus gained hydrogenase genes and genes for ferredoxin reduction that appear to permit syntrophic growth via hydrogen production. The gain of new physiological capabilities in the Pelobacter species were accompanied by the loss of several key genes necessary for the complete oxidation of organic compounds and the genes for the c-type cytochromes required for extracellular electron transfer. CONCLUSION: The results suggest that Pelobacter species evolved parallel strategies to enhance their ability to compete in environments in which electron acceptors for anaerobic respiration were limiting. More generally, these results demonstrate how relatively few gene changes can dramatically transform metabolic capabilities and expand the range of environments in which microorganisms can compete. PMID- 19284580 TI - Diversity and transcription of proteases involved in the maturation of hydrogenases in Nostoc punctiforme ATCC 29133 and Nostoc sp. strain PCC 7120. AB - BACKGROUND: The last step in the maturation process of the large subunit of [NiFe]-hydrogenases is a proteolytic cleavage of the C-terminal by a hydrogenase specific protease. Contrary to other accessory proteins these hydrogenase proteases are believed to be specific whereby one type of hydrogenases specific protease only cleaves one type of hydrogenase. In cyanobacteria this is achieved by the gene product of either hupW or hoxW, specific for the uptake or the bidirectional hydrogenase respectively. The filamentous cyanobacteria Nostoc punctiforme ATCC 29133 and Nostoc sp strain PCC 7120 may contain a single uptake hydrogenase or both an uptake and a bidirectional hydrogenase respectively. RESULTS: In order to examine these proteases in cyanobacteria, transcriptional analyses were performed of hupW in Nostoc punctiforme ATCC 29133 and hupW and hoxW in Nostoc sp. strain PCC 7120. These studies revealed numerous transcriptional start points together with putative binding sites for NtcA (hupW) and LexA (hoxW). In order to investigate the diversity and specificity among hydrogeanse specific proteases we constructed a phylogenetic tree which revealed several subgroups that showed a striking resemblance to the subgroups previously described for [NiFe]-hydrogenases. Additionally the proteases specificity was also addressed by amino acid sequence analysis and protein-protein docking experiments with 3D-models derived from bioinformatic studies. These studies revealed a so called "HOXBOX"; an amino acid sequence specific for protease of Hox-type which might be involved in docking with the large subunit of the hydrogenase. CONCLUSION: Our findings suggest that the hydrogenase specific proteases are under similar regulatory control as the hydrogenases they cleave. The result from the phylogenetic study also indicates that the hydrogenase and the protease have co-evolved since ancient time and suggests that at least one major horizontal gene transfer has occurred. This co-evolution could be the result of a close interaction between the protease and the large subunit of the [NiFe]-hydrogenases, a theory supported by protein-protein docking experiments performed with 3D-models. Finally we present data that may explain the specificity seen among hydrogenase specific proteases, the so called "HOXBOX"; an amino acid sequence specific for proteases of Hox-type. This opens the door for more detailed studies of the specificity found among hydrogenase specific proteases and the structural properties behind it. PMID- 19284581 TI - Characterization of the hupSL promoter activity in Nostoc punctiforme ATCC 29133. AB - BACKGROUND: In cyanobacteria three enzymes are directly involved in the hydrogen metabolism; a nitrogenase that produces molecular hydrogen, H2, as a by-product of nitrogen fixation, an uptake hydrogenase that recaptures H2 and oxidize it, and a bidirectional hydrogenase that can both oxidize and produce H2.Nostoc punctiforme ATCC 29133 is a filamentous dinitrogen fixing cyanobacterium containing a nitrogenase and an uptake hydrogenase but no bidirectional hydrogenase. Generally, little is known about the transcriptional regulation of the cyanobacterial uptake hydrogenases. In this study gel shift assays showed that NtcA has a specific affinity to a region of the hupSL promoter containing a predicted NtcA binding site. The predicted NtcA binding site is centred at 258.5 bp upstream the transcription start point (tsp). To further investigate the hupSL promoter, truncated versions of the hupSL promoter were fused to either gfp or luxAB, encoding the reporter proteins Green Fluorescent Protein and Luciferase, respectively. RESULTS: Interestingly, all hupsSL promoter deletion constructs showed heterocyst specific expression. Unexpectedly the shortest promoter fragment, a fragment covering 57 bp upstream and 258 bp downstream the tsp, exhibited the highest promoter activity. Deletion of the NtcA binding site neither affected the expression to any larger extent nor the heterocyst specificity. CONCLUSION: Obtained data suggest that the hupSL promoter in N. punctiforme is not strictly dependent on the upstream NtcA cis element and that the shortest promoter fragment (-57 to tsp) is enough for a high and heterocyst specific expression of hupSL. This is highly interesting because it indicates that the information that determines heterocyst specific gene expression might be confined to this short sequence or in the downstream untranslated leader sequence. PMID- 19284582 TI - Toxicogenomic analysis of susceptibility to inhaled urban particulate matter in mice with chronic lung inflammation. AB - BACKGROUND: Individuals with chronic lung disease are at increased risk of adverse health effects from airborne particulate matter. Characterization of underlying pollutant-phenotype interactions may require comprehensive strategies. Here, a toxicogenomic approach was used to investigate how inflammation modifies the pulmonary response to urban particulate matter. RESULTS: Transgenic mice with constitutive pulmonary overexpression of tumour necrosis factor (TNF)-alpha under the control of the surfactant protein C promoter and wildtype littermates (C57BL/6 background) were exposed by inhalation for 4 h to particulate matter (0 or 42 mg/m3 EHC-6802) and euthanized 0 or 24 h post-exposure. The low alveolar dose of particles (16 mug) did not provoke an inflammatory response in the lungs of wildtype mice, nor exacerbate the chronic inflammation in TNF animals. Real time PCR confirmed particle-dependent increases of CYP1A1 (30-100%), endothelin-1 (20-40%), and metallothionein-II (20-40%) mRNA in wildtype and TNF mice (p < 0.05), validating delivery of a biologically-effective dose. Despite detection of striking genotype-related differences, including activation of immune and inflammatory pathways consistent with the TNF-induced pathology, and time-related effects attributable to stress from nose-only exposure, microarray analysis failed to identify effects of the inhaled particles. Remarkably, the presence of chronic inflammation did not measurably amplify the transcriptional response to particulate matter. CONCLUSION: Our data support the hypothesis that health effects of acute exposure to urban particles are dominated by activation of specific physiological response cascades rather than widespread changes in gene expression. PMID- 19284583 TI - MASEP gamma knife radiosurgery for secretory pituitary adenomas: experience in 347 consecutive cases. AB - BACKGROUND: Secretory pituitary adenomas are very common brain tumors. Historically, the treatment armamentarium for secretory pituitary adenomas included neurosurgery, medical management, and fractionated radiotherapy. In recent years, MASEP gamma knife radiosurgery (MASEP GKRS) has emerged as an important treatment modality in the management of secretory pituitary adenomas. The goal of this research is to define accurately the efficacy, safety, complications, and role of MASEP GKRS for treatment of secretory pituitary adenomas. METHODS: Between 1997 and 2007 a total of 347 patients with secretory pituitary adenomas treated with MASEP GKRS and with at least 60 months of follow up data were identified. In 47 of these patients some form of prior treatment such as transsphenoidal resection, or craniotomy and resection had been conducted. The others were deemed ineligible for microsurgery because of body health or private choice, and MASEP GKRS served as the primary treatment modality. Endocrinological, ophthalmological, and neuroradiological responses were evaluated. RESULTS: MASEP GKRS was tolerated well in these patients under the follow-up period ranged from 60 to 90 months; acute radioreaction was rare and 17 patients had transient headaches with no clinical significance. Late radioreaction was noted in 1 patient and consisted of consistent headache. Of the 68 patients with adrenocorticotropic hormone-secreting(ACTH) adenomas, 89.7% showed tumor volume decrease or remain unchanged and 27.9% experienced normalization of hormone level. Of the 176 patients with prolactinomas, 23.3% had normalization of hormone level and 90.3% showed tumor volume decrease or remain unchanged. Of the 103 patients with growth hormone-secreting(GH) adenomas, 95.1% experienced tumor volume decrease or remain unchanged and 36.9% showed normalization of hormone level. CONCLUSION: MASEP GKRS is safe and effective in treating secretory pituitary adenomas. None of the patients in our study experienced injury to the optic apparatus or had other neuropathies related with gamma knife. MASEP GKRS may serve as a primary treatment method in some or as a salvage treatment in the others. However, treatment must be tailored to meet the patient's symptoms, tumor location, tumor morphometry, and overall health. Longer follow-up is required for a more complete assessment of late radioreaction and treatment efficacy. PMID- 19284584 TI - Identification of conformational epitopes for human IgG on Chemotaxis inhibitory protein of Staphylococcus aureus. AB - BACKGROUND: The Chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) blocks the Complement fragment C5a receptor (C5aR) and formylated peptide receptor (FPR) and is thereby a potent inhibitor of neutrophil chemotaxis and activation of inflammatory responses. The majority of the healthy human population has antibodies against CHIPS that have been shown to interfere with its function in vitro. The aim of this study was to define potential epitopes for human antibodies on the CHIPS surface. We also initiate the process to identify a mutated CHIPS molecule that is not efficiently recognized by preformed anti-CHIPS antibodies and retains anti-inflammatory activity. RESULTS: In this paper, we panned peptide displaying phage libraries against a pool of CHIPS specific affinity-purified polyclonal human IgG. The selected peptides could be divided into two groups of sequences. The first group was the most dominant with 36 of the 48 sequenced clones represented. Binding to human affinity-purified IgG was verified by ELISA for a selection of peptide sequences in phage format. For further analysis, one peptide was chemically synthesized and antibodies affinity purified on this peptide were found to bind the CHIPS molecule as studied by ELISA and Surface Plasmon Resonance. Furthermore, seven potential conformational epitopes responsible for antibody recognition were identified by mapping phage selected peptide sequences on the CHIPS surface as defined in the NMR structure of the recombinant CHIPS31-121 protein. Mapped epitopes were verified by in vitro mutational analysis of the CHIPS molecule. Single mutations introduced in the proposed antibody epitopes were shown to decrease antibody binding to CHIPS. The biological function in terms of C5aR signaling was studied by flow cytometry. A few mutations were shown to affect this biological function as well as the antibody binding. CONCLUSION: Conformational epitopes recognized by human antibodies have been mapped on the CHIPS surface and amino acid residues involved in both antibody and C5aR interaction could be defined. This information has implications for the development of an effective anti-inflammatory agent based on a functional CHIPS molecule with low interaction with human IgG. PMID- 19284585 TI - Is parental coping associated with quality of life in juvenile idiopathic arthritis? AB - Parents of children with a chronic condition such as juvenile arthritis must cope with greater demands than those living with a healthy child. They must adopt different behaviours in order to lessen the impact on the family structure. Parental coping refers to the parent's specific cognitive and behavioural efforts to reduce or manage a demand on the family system. The aims of this study were: to describe coping in a cohort of parents of children with JIA; to determine whether quality of life is associated with parental coping; to explore whether socio-demographic factors such as child's age, family socioeconomic status and family structure are associated with parental coping. One hundred eighty-two parents caring for a child with JIA completed a postal survey at three times over a one-year period, which included the Juvenile Arthritis Quality of Life Questionnaire (JAQQ), the Coping Health Inventory for Parents (CHIP) and questionnaires describing socio-demographic characteristics. Linear mixed models were employed to analyse the association between the child's quality of life and parental coping. Mean total QoL scores (JAQQ) showed that children experienced difficulty in completing specified activities at most just below 25% of the time and results fall off slightly following the 6 month time point. Mean parental coping scores for the CHIP subscales at baseline were 38.4 +/- 9.0, 33.4 +/- 11.6, 16.5 +/- 6.1, for Maintaining Family Integration (maximum score 57), Maintaining Social Support (maximum score 54) and Understanding the Medical Situation (maximum score 24), respectively. Understanding the Medical Situation was deemed most useful. The child's QoL was associated with parental coping. Parents of children with greater psychosocial dysfunction used more coping behaviours related to Understanding the Medical Situation (beta coefficient, 0.73; 95% CI, 0.01, 1.45). These findings underscore the importance of helping parents of children with JIA better understand their child's medical situation. PMID- 19284586 TI - Synthesis of rhamnolipid biosurfactant and mode of hexadecane uptake by Pseudomonas species. AB - BACKGROUND: Microorganisms have devised ways by which they increase the bioavailability of many water immiscible substrates whose degradation rates are limited by their low water solubility. Hexadecane is one such water immiscible hydrocarbon substrate which forms an important constituent of oil. One major mechanism employed by hydrocarbon degrading organisms to utilize such substrates is the production of biosurfactants. However, much of the overall mechanism by which such organisms utilize hydrocarbon substrate still remains a mystery. RESULTS: With an aim to gain more insight into hydrocarbon uptake mechanism, an efficient biosurfactant producing and n-hexadecane utilizing Pseudomonas sp was isolated from oil contaminated soil which was found to produce rhamnolipid type of biosurfactant containing a total of 13 congeners. Biosurfactant action brought about the dispersion of hexadecane to droplets smaller than 0.22 mum increasing the availability of the hydrocarbon to the degrading organism. Involvement of biosurfactant was further confirmed by electron microscopic studies. Biosurfactant formed an emulsion with hexadecane thereby facilitating increased contact between hydrocarbon and the degrading bacteria. Interestingly, it was observed that "internalization" of "biosurfactant layered hydrocarbon droplet" was taking place suggesting a mechanism similar in appearance to active pinocytosis, a fact not earlier visually reported in bacterial systems for hydrocarbon uptake. CONCLUSION: This study throws more light on the uptake mechanism of hydrocarbon by Pseudomonas aeruginosa. We report here a new and exciting line of research for hydrocarbon uptake involving internalization of biosurfactant covered hydrocarbon inside cell for subsequent breakdown. PMID- 19284587 TI - High-throughput mutational analysis of TOR1A in primary dystonia. AB - BACKGROUND: Although the c.904_906delGAG mutation in Exon 5 of TOR1A typically manifests as early-onset generalized dystonia, DYT1 dystonia is genetically and clinically heterogeneous. Recently, another Exon 5 mutation (c.863G>A) has been associated with early-onset generalized dystonia and some DeltaGAG mutation carriers present with late-onset focal dystonia. The aim of this study was to identify TOR1A Exon 5 mutations in a large cohort of subjects with mainly non generalized primary dystonia. METHODS: High resolution melting (HRM) was used to examine the entire TOR1A Exon 5 coding sequence in 1014 subjects with primary dystonia (422 spasmodic dysphonia, 285 cervical dystonia, 67 blepharospasm, 41 writer's cramp, 16 oromandibular dystonia, 38 other primary focal dystonia, 112 segmental dystonia, 16 multifocal dystonia, and 17 generalized dystonia) and 250 controls (150 neurologically normal and 100 with other movement disorders). Diagnostic sensitivity and specificity were evaluated in an additional 8 subjects with known DeltaGAG DYT1 dystonia and 88 subjects with DeltaGAG-negative dystonia. RESULTS: HRM of TOR1A Exon 5 showed high (100%) diagnostic sensitivity and specificity. HRM was rapid and economical. HRM reliably differentiated the TOR1A DeltaGAG and c.863G>A mutations. Melting curves were normal in 250/250 controls and 1012/1014 subjects with primary dystonia. The two subjects with shifted melting curves were found to harbor the classic DeltaGAG deletion: 1) a non-Jewish Caucasian female with childhood-onset multifocal dystonia and 2) an Ashkenazi Jewish female with adolescent-onset spasmodic dysphonia. CONCLUSION: First, HRM is an inexpensive, diagnostically sensitive and specific, high throughput method for mutation discovery. Second, Exon 5 mutations in TOR1A are rarely associated with non-generalized primary dystonia. PMID- 19284588 TI - Lipopolysaccharide-induced interleukin-6 production is controlled by glycogen synthase kinase-3 and STAT3 in the brain. AB - BACKGROUND: Septic shock is a prevalent condition that, when not lethal, often causes disturbances in cognition, mood, and behavior, particularly due to central actions of the inflammatory cytokine interleukin-6 (IL-6). To identify potential targets to control brain IL-6, we tested if IL-6 produced by glia is regulated by signal transducer and activator of transcription-3 (STAT3) and glycogen synthase kinase-3 (GSK3). METHODS: Lipopolysaccharide (LPS) was used to induce inflammatory responses in mice or cultured primary glia. IL-6 was measured by ELISA and other inflammatory molecules were measured using an array. RESULTS: Mouse brain IL-6 levels increased after central, as well as peripheral, LPS administration, consistent with glia producing a portion of brain IL-6. STAT3 in the brain was activated after peripheral or central LPS administration, and in LPS-stimulated cultured primary glia. Inhibition of STAT3 expression, function, or activation reduced by ~80% IL-6 production by primary glia, demonstrating the dependence on active STAT3. GSK3 promotes STAT3 activation, and array analysis of inflammatory molecules produced by LPS-stimulated primary glia demonstrated that IL-6 was the cytokine most diminished (>90%) by GSK3 inhibition. Inhibition of GSK3, and knockdown of GSK3beta, not GSK3alpha, greatly inhibited IL-6 production by LPS-stimulated primary glia. Conversely, expression of active STAT3 and active GSK3 promoted IL-6 production. In vivo inhibition of GSK3 reduced serum and brain IL-6 levels, brain STAT3 activation, and GFAP upregulation following LPS administration. CONCLUSION: STAT3 and GSK3 cooperatively promote neuroinflammation, providing novel targets for anti-inflammatory intervention. PMID- 19284589 TI - Resistance training with soy vs whey protein supplements in hyperlipidemic males. AB - BACKGROUND: Most individuals at risk for developing cardiovascular disease (CVD) can reduce risk factors through diet and exercise before resorting to drug treatment. The effect of a combination of resistance training with vegetable based (soy) versus animal-based (whey) protein supplementation on CVD risk reduction has received little study. The study's purpose was to examine the effects of 12 weeks of resistance exercise training with soy versus whey protein supplementation on strength gains, body composition and serum lipid changes in overweight, hyperlipidemic men. METHODS: Twenty-eight overweight, male subjects (BMI 25-30) with serum cholesterol >200 mg/dl were randomly divided into 3 groups (placebo (n = 9), and soy (n = 9) or whey (n = 10) supplementation) and participated in supervised resistance training for 12 weeks. Supplements were provided in a double blind fashion. RESULTS: All 3 groups had significant gains in strength, averaging 47% in all major muscle groups and significant increases in fat free mass (2.6%), with no difference among groups. Percent body fat and waist-to-hip ratio decreased significantly in all 3 groups an average of 8% and 2%, respectively, with no difference among groups. Total serum cholesterol decreased significantly, again with no difference among groups. CONCLUSION: Participation in a 12 week resistance exercise training program significantly increased strength and improved both body composition and serum cholesterol in overweight, hypercholesterolemic men with no added benefit from protein supplementation. PMID- 19284590 TI - Genomic scan for quantitative trait loci of chemical and physical body composition and deposition on pig chromosome X including the pseudoautosomal region of males. AB - A QTL analysis of pig chromosome X (SSCX) was carried out using an approach that accurately takes into account the specific features of sex chromosomes i.e. their heterogeneity, the presence of a pseudoautosomal region and the dosage compensation phenomenon. A three-generation full-sib population of 386 animals was created by crossing Pietrain sires with a crossbred dam line. Phenotypic data on 72 traits were recorded for at least 292 and up to 315 F2 animals including chemical body composition measured on live animals at five target weights ranging from 30 to 140 kg, daily gain and feed intake measured throughout growth, and carcass characteristics obtained at slaughter weight (140 kg). Several significant and suggestive QTL were detected on pig chromosome X: (1) in the pseudoautosomal region of SSCX, a QTL for entire loin weight, which showed paternal imprinting, (2) closely linked to marker SW2456, a suggestive QTL for feed intake at which Pietrain alleles were found to be associated with higher feed intake, which is unexpected for a breed known for its low feed intake capacity, (3) at the telomeric end of the q arm of SSCX, QTL for jowl weight and lipid accretion and (4) suggestive QTL for chemical body composition at 30 kg. These results indicate that SSCX is important for physical and chemical body composition and accretion as well as feed intake regulation. PMID- 19284592 TI - Genetic diversity of msp3alpha and msp1_b5 markers of Plasmodium vivax in French Guiana. AB - BACKGROUND: Reliable molecular typing tools are required for a better understanding of the molecular epidemiology of Plasmodium vivax. The genes msp3a and msp1_block5 are highly polymorphic and have been used as markers in many P. vivax population studies. These markers were used to assess the genetic diversity of P. vivax strains from French Guiana (South America) and to develop a molecular typing protocol. METHODS: A total of 120 blood samples from 109 patients (including 10 patients suffered from more than one malaria episode, samples were collected during each episode) with P. vivax infection were genotyped. All samples were analysed by msp3a PCR-RFLP and msp1_b5 gene sequencing was performed on 57 samples. Genotyping protocol applied to distinguish between new infection or relapse from heterologus hypnozoites and treatment failure or relapse from homologus hypnozoites was based on analysing first msp3a by PCR-RFLP and secondly, only if the genotypes of the two samples are identical, on sequencing the msp1_b5 gene. RESULTS: msp3a alleles of three sizes were amplified by PCR: types A, B and C. Eleven different genotypes were identified among the 109 samples analysed by msp3a PCR-RFLP. In 13.8% of cases, a mixed genotype infection was observed. The sequence of msp1_b5 gene revealed 22 unique genotypes and 12.3% of cases with mixed infection. In the 57 samples analysed by both methods, 45 genotypes were found and 21% were mixed. Among ten patients with two or three malaria episodes, the protocol allowed to identify five new infections or relapses from heterologous hypnozoites and six treatment failures of relapses from homologous hypnozoites. CONCLUSION: The study showed a high diversity of msp3a and msp1_b5 genetic markers among P. vivax strains in French Guiana with a low polyclonal infection rate. These results indicated that the P. vivax genotyping protocol presented has a good discrimination power and can be used in clinical drug trials or epidemiological studies. PMID- 19284591 TI - Management of burn injuries--recent developments in resuscitation, infection control and outcomes research. AB - INTRODUCTION: Burn injury and its subsequent multisystem effects are commonly encountered by acute care practitioners. Resuscitation is the major component of initial burn care and must be managed to restore and preserve remote organ function. Later complications of burn injury are dominated by infection. Burn centers are often called to manage soft tissue problems outside thermal injury including soft tissue infection and Toxic Epidermal Necrolysis. METHODS: A selected review of recent reports published by the American Burn Association is provided. RESULTS: The burn-injured patient is easily and frequently over resuscitated with complications including delayed wound healing and respiratory compromise. A feedback protocol is designed to limit the occurrence of excessive resuscitation has been proposed but no new "gold standard" for resuscitation has replaced the Parkland formula. Significant additional work has been included in recent guidelines identifying specific infectious complications and criteria for these diagnoses in the burn-injured patient. While new medical therapies have been proposed for patients sustaining inhalation injury, a new standard of medical therapy has not emerged. Renal failure as a contributor to adverse outcome in burns has been reinforced by recent data generated in Scandinavia. Of special problems addressed in burn centers, soft tissue infections and Toxic Epidermal Necrolysis have been reviewed but new treatment strategies have not been identified. The value of burn centers in management of burns and other soft tissue problems is supported in several recent reports. CONCLUSION: Recent reports emphasize the dangers of over resuscitation in the setting of burn injury. No new medical therapy for inhalation injury exists but new standards for description of burn-related infections have been presented. The value of the burn center in care of soft tissue problems including Toxic Epidermal Necrolysis and soft tissue infections is supported in recent papers. PMID- 19284593 TI - Mapping the history and current situation of research on John Cunningham virus - a bibliometric analysis. AB - BACKGROUND: John Cunningham virus (JCV) constitutes a family of polyoma viruses, which plays important roles in the progressive multifocal leukoencephalopathy (PML) and tumorigenesis. However, no bibliometric investigation has been reported to guide the researchers and potential readers. METHODS: Papers were collected from database Sci-expanded and Pubmed until May 22, 2008. The highly-productive authors, institutes and countries, highly-cited authors and journals were ranked. The highly-cited articles were subjected to co-citation and chronological analysis with highly-frequent MeSH words for co-occurrence analysis. RESULTS: Until now, 1785 articles about JCV were indexed in Sci-expanded and 1506 in Pubmed. The main document type was original article. USA, Japan and Italy were the largest three producers about JCV. Temple University published 128 papers and ranked the top, followed by University of Tokyo. Khalili K and Yogo Y became the core authors due to more than 20 documents produced. Journal of Neurovirology published more than 15 papers and ranked the top. Padgett BL and Berger JR were the first two highly-cited authors. Journal of Virology and Journal of Neurovirology respectively ranked to the first two highly-cited journals. These top highly-cited articles were divided into 5 aspects: (1) The correlation between JC virus and tumors; (2) Causal correlation of JCV with PML; (3) Polyoma virus infection and its related diseases in renal-allograft recipients; (4) Detection of JCV antibody, oncogene and its encoding protein; (5) Genetics and molecular biology of JCV. The MeSH/subheadings were classified into five groups: (1) JCV and virus infectious diseases; (2) JCV pathogenicity and pathological appearance of PML; (3) JCV isolation and detection; (4) Immunology of JCV and PML; (5) JCV genetics and tumors. CONCLUSION: JCV investigation mainly focused on its isolation and detection, as well as its correlation with PML and tumors. Establishment of transgenic animal model using JCV T antigen would be a hopeful and useful project in the further study. PMID- 19284594 TI - High sensitivity detection of Plasmodium species reveals positive correlations between infections of different species, shifts in age distribution and reduced local variation in Papua New Guinea. AB - BACKGROUND: When diagnosed by standard light microscopy (LM), malaria prevalence can vary significantly between sites, even at local scale, and mixed species infections are consistently less common than expect in areas co-endemic for Plasmodium falciparum, Plasmodium vivax and Plasmodium malariae. The development of a high-throughput molecular species diagnostic assay now enables routine PCR based surveillance of malaria infections in large field and intervention studies, and improves resolution of species distribution within and between communities. METHODS: This study reports differences in the prevalence of infections with all four human malarial species and of mixed infections as diagnosed by LM and post PCR ligase detection reaction-fluorescent microsphere (LDR-FMA) assay in 15 villages in the central Sepik area of Papua New Guinea. RESULTS: Significantly higher rates of infection by P. falciparum, P. vivax, P. malariae and Plasmodium ovale were observed in LDR-FMA compared to LM diagnosis (p < 0.001). Increases were particularly pronounced for P. malariae (3.9% vs 13.4%) and P. ovale (0.0% vs 4.8%). In contrast to LM diagnosis, which suggested a significant deficit of mixed species infections, a significant excess of mixed infections over expectation was detected by LDR-FMA (p < 0.001). Age of peak prevalence shifted to older age groups in LDR-FMA diagnosed infections for P. falciparum (LM: 7-9 yrs 47.5%, LDR-FMA: 10-19 yrs 74.2%) and P. vivax (LM: 4-6 yrs 24.2%, LDR-FMA: 7 9 yrs 50.9%) but not P. malariae infections (10-19 yrs, LM: 7.7% LDR-FMA: 21.6%). Significant geographical variation in prevalence was found for all species (except for LM-diagnosed P. falciparum), with the extent of this variation greater in LDR-FMA than LM diagnosed infections (overall, 84.4% vs. 37.6%). Insecticide-treated bednet (ITN) coverage was also the dominant factor linked to geographical differences in Plasmodium species infection prevalence explaining between 60.6% - 74.5% of this variation for LDR-FMA and 81.8% - 90.0% for LM (except P. falciparum), respectively. CONCLUSION: The present study demonstrates that application of molecular diagnosis reveals patterns of malaria risk that are significantly different from those obtained by standard LM. Results provide insight relevant to design of malaria control and eradication strategies. PMID- 19284595 TI - An optimised system for refolding of human glucose 6-phosphate dehydrogenase. AB - BACKGROUND: Human glucose 6-phosphate dehydrogenase (G6PD), active in both dimer and tetramer forms, is the key entry enzyme in the pentose phosphate pathway (PPP), providing NADPH for biosynthesis and various other purposes, including protection against oxidative stress in erythrocytes. Accordingly haemolytic disease is a major consequence of G6PD deficiency mutations in man, and many severe disease phenotypes are attributed to G6PD folding problems. Therefore, a robust refolding method with high recovery yield and reproducibility is of particular importance to study those clinical mutant enzymes as well as to shed light generally on the refolding process of large multi-domain proteins. RESULTS: The effects of different chemical and physical variables on the refolding of human recombinant G6PD have been extensively investigated. L-Arg, NADP+ and DTT are all major positive influences on refolding, and temperature, protein concentration, salt types and other additives also have significant impacts. With the method described here, ~70% enzyme activity could be regained, with good reproducibility, after denaturation with Gdn-HCl, by rapid dilution of the protein, and the refolded enzyme displays kinetic and CD properties indistinguishable from those of the native protein. Refolding under these conditions is relatively slow, taking about 7 days to complete at room temperature even in the presence of cyclophilin A, a peptidylprolyl isomerase reported to increase refolding rates. The refolded protein intermediates shift from dominant monomer to dimer during this process, the gradual emergence of dimer correlating well with the regain of enzyme activity. CONCLUSION: L-Arg is the key player in the refolding of human G6PD, preventing the aggregation of folding intermediate, and NADP+ is essential for the folding intermediate to adopt native structure. The refolding protocol can be applied to produce high recovery yield of folded protein with unaltered properties, paving the way for future studies on clinical G6PD mutants with folding defects and providing a useful model system to study the folding process of oligomeric proteins. PMID- 19284596 TI - A comparative analysis of divergently-paired genes (DPGs) among Drosophila and vertebrate genomes. AB - BACKGROUND: Divergently-paired genes (DPGs) are defined as two adjacent genes that are transcribed toward the opposite direction (or from different DNA strands) and shared their transcription start sites (TSSs) less than 1,000 base pairs apart. DPGs are products of a common organizational feature among eukaryotic genes yet to be surveyed across divergent genomes over well-defined evolutionary distances since mutations in the sequence between a pair of DPGs may result in alternations in shared promoters and thus affect the function of both genes. By sharing promoters, the gene pairs take the advantage of co-regulation albeit bearing doubled mutational burdens in maintaining their normal functions. RESULTS: Drosophila melanogaster has a significant fraction (31.6% of all genes) of DPGs which are remarkably conserved relative to its gene density as compared to other eukaryotes. Our survey and comparative analysis revealed different evolutionary patterns among DPGs between insect and vertebrate lineages. The conservation of DPGs in D. melanogaster is of significance as they are mostly housekeeping genes characterized by the absence of TATA box in their promoter sequences. The combination of Initiator and Downstream Promoter Element may play an important role in regulating DPGs in D. melanogaster, providing an excellent niche for studying the molecular details for transcription regulations. CONCLUSION: DPGs appear to have arisen independently among different evolutionary lineages, such as the insect and vertebrate lineages, and exhibit variable degrees of conservation. Such architectural organizations, including convergently paired genes (CPGs) may associate with transcriptional regulation and have significant functional relevance. PMID- 19284598 TI - Gender-specific epidemiology of diabetes: a representative cross-sectional study. AB - BACKGROUND: Diabetes and its associated complications are part of a chronic disease global epidemic that presents a public health challenge. Epidemiologists examining health differences between men and women are being challenged to recognise the biological and social constructions behind the terms 'sex' and/or 'gender', together with social epidemiology principles and the life course approach. This paper examines the epidemiology of a population with diabetes from the north-west metropolitan region of South Australia. METHODS: Data were used from a sub-population with diabetes (n = 263), from 4060 adults aged 18 years and over living in the north-west suburbs of Adelaide, South Australia. Eligible respondents were asked to participate in a telephone interview, a self-report questionnaire and a biomedical examination. Diabetes (undiagnosed and diagnosed) was determined using self-reported information and a fasting blood test administered to participants. Data were analysed using SPSS (Version 10.0) and EpiInfo (Version 6.0). RESULTS: Factors associated with diabetes for both men and women were being aged 40 years and over, and having a low gross annual household income, obesity and a family history of diabetes. In addition, being an ex-smoker and having low cholesterol levels were associated with diabetes among men. Among women, having a high waist-hip ratio, high blood pressure and reporting a previous cardiovascular event or mental health problem were associated with diabetes. CONCLUSION: The results found that men and women with diabetes face different challenges in the management of their condition. Public health implications include a need for quality surveillance data, including epidemiological life course, social, behavioural, genetic and environmental factors. This will enrich the evidence base for health promotion professionals and allow policy makers to draw inferences and conclusions for interventions and planning purposes. PMID- 19284597 TI - The role of tau in neurodegeneration. AB - Since the identification of tau as the main component of neurofibrillary tangles in Alzheimer's disease and related tauopathies, and the discovery that mutations in the tau gene cause frontotemporal dementia, much effort has been directed towards determining how the aggregation of tau into fibrillar inclusions causes neuronal death. As evidence emerges that tau-mediated neuronal death can occur even in the absence of tangle formation, a growing number of studies are focusing on understanding how abnormalities in tau (e.g. aberrant phosphorylation, glycosylation or truncation) confer toxicity. Though data obtained from experimental models of tauopathies strongly support the involvement of pathologically modified tau and tau aggregates in neurodegeneration, the exact neurotoxic species remain unclear, as do the mechanism(s) by which they cause neuronal death. Nonetheless, it is believed that tau-mediated neurodegeneration is likely to result from a combination of toxic gains of function as well as from the loss of normal tau function. To truly appreciate the detrimental consequences of aberrant tau function, a better understanding of all functions carried out by tau, including but not limited to the role of tau in microtubule assembly and stabilization, is required. This review will summarize what is currently known regarding the involvement of tau in the initiation and development of neurodegeneration in tauopathies, and will also highlight some of the remaining questions in need of further investigation. PMID- 19284599 TI - Comparison of markers of oxidative stress, inflammation and arterial stiffness between incident hemodialysis and peritoneal dialysis patients--an observational study. AB - BACKGROUND: Patients on peritoneal and hemodialysis have accelerated atherosclerosis associated with an increase in cardiovascular morbidity and mortality. The atherosclerosis is associated with increased arterial stiffness, endothelial dysfunction and elevated oxidative stress and inflammation. The aims of this study are to investigate the effects of peritoneal and hemodialysis on arterial stiffness, vascular function, myocardial structure and function, oxidative stress and inflammation in incident patients with end stage kidney disease. METHODS: This is an observational study. Eighty stage five CKD patients will be enrolled and followed for one-year. Primary outcome measures will be changes in 1) arterial stiffness measured by aortic pulse wave velocity, 2) oxidative stress assessed by plasma F2 isoprostanes and 3) inflammation measured by plasma pentraxin-3. Secondary outcomes will include additional measures of oxidative stress and inflammation, changes in vascular function assessed using the brachial artery reactivity technique, carotid artery intimal medial thickness, augmentation index and trans thoracic echocardiography to assess left ventricular geometry, and systolic and diastolic function. Patients will undergo these measures at baseline (6-8 weeks prior to starting dialysis therapy), then at six and 12 months after starting dialysis. DISCUSSION: The results of this study may guide the choice of dialysis modality in the first year of treatment. It may also lead to a larger study prospectively assessing the effect of dialysis modality on cardiovascular morbidity and mortality. TRIAL REGISTRATION: ACTRN12609000049279. PMID- 19284600 TI - Spatiotemporal dipole source localization of face processing ERPs in adolescents: a preliminary study. AB - BACKGROUND: Despite extensive investigation of the neural systems for face perception and emotion recognition in adults and young children in the past, the precise temporal activation of brain sources specific to the processing of emotional facial expressions in older children and adolescents is not well known. This preliminary study aims to trace the spatiotemporal dynamics of facial emotion processing during adolescence and provide a basis for future developmental studies and comparisons with patient populations that have social emotional deficits such as autism. METHODS: We presented pictures showing happy, angry, fearful, or neutral facial expressions to healthy adolescents (aged 10-16 years) and recorded 128-channel event-related potentials (ERPs) while they performed an emotion discrimination task. ERP components were analyzed for effects of age and emotion on amplitude and latency. The underlying cortical sources of scalp ERP activity were modeled as multiple equivalent current dipoles using Brain Electrical Source Analysis (BESA). RESULTS: Initial global/holistic processing of faces (P1) took place in the visual association cortex (lingual gyrus) around 120 ms post-stimulus. Next, structural encoding of facial features (N170) occurred between 160-200 ms in the inferior temporal/fusiform region, and perhaps early emotion processing (Vertex Positive Potential or VPP) in the amygdala and orbitofrontal cortex. Finally, cognitive analysis of facial expressions (P2) in the prefrontal cortex and emotional reactions in somatosensory areas were observed from about 230 ms onwards. The temporal sequence of cortical source activation in response to facial emotion processing was occipital, prefrontal, fusiform, parietal for young adolescents and occipital, limbic, inferior temporal, and prefrontal for older adolescents. CONCLUSION: This is a first report of high-density ERP dipole source analysis in healthy adolescents which traces the sequence of neural activity within the first 500 ms of categorizing emotion from faces. Our spatio-temporal brain source models showed the presence of adult-like cortical networks for face processing in adolescents, whose functional specificity to different emotions appear to be not yet fully mature. Age-related differences in brain activation patterns illustrate the continued development and maturation of distinct neural systems for processing facial expressions during adolescence and possible changes in emotion perception, experience, and reaction with age. PMID- 19284601 TI - Novel autoantigens immunogenic in COPD patients. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a respiratory inflammatory condition with autoimmune features including IgG autoantibodies. In this study we analyze the complexity of the autoantibody response and reveal the nature of the antigens that are recognized by autoantibodies in COPD patients. METHODS: An array of 1827 gridded immunogenic peptide clones was established and screened with 17 sera of COPD patients and 60 healthy controls. Protein arrays were evaluated both by visual inspection and a recently developed computer aided image analysis technique. By this computer aided image analysis technique we computed the intensity values for each peptide clone and each serum and calculated the area under the receiver operator characteristics curve (AUC) for each clone and the separation COPD sera versus control sera. RESULTS: By visual evaluation we detected 381 peptide clones that reacted with autoantibodies of COPD patients including 17 clones that reacted with more than 60% of the COPD sera and seven clones that reacted with more than 90% of the COPD sera. The comparison of COPD sera and controls by the automated image analysis system identified 212 peptide clones with informative AUC values. By in silico sequence analysis we found an enrichment of sequence motives previously associated with immunogenicity. CONCLUSION: The identification of a rather complex humoral immune response in COPD patients supports the idea of COPD as a disease with strong autoimmune features. The identification of novel immunogenic antigens is a first step towards a better understanding of the autoimmune component of COPD. PMID- 19284602 TI - Adam33 polymorphisms are associated with COPD and lung function in long-term tobacco smokers. AB - BACKGROUND: Variation in ADAM33 has been shown to be important in the development of asthma and altered lung function. This relationship however, has not been investigated in the population susceptible to COPD; long term tobacco smokers. We evaluated the association between polymorphisms in ADAM33 gene with COPD and lung function in long term tobacco smokers. METHODS: Caucasian subjects, at least 50 year old, who smoked >or= 20 pack-years (n = 880) were genotyped for 25 single nucleotide polymorphisms (SNPs) in ADAM33. COPD was defined as an FEV1/FVC ratio < 70% and percent-predicted (pp)FEV1 < 75% (n = 287). The control group had an FEV1/FVC ratio >or= 70% and ppFEV(1) >or= 80% (n = 311) despite >or= 20 pack years of smoking. Logistic and linear regressions were used for the analysis. Age, sex, and smoking status were considered as potential confounders. RESULTS: Five SNPs in ADAM33 were associated with COPD (Q-1, intronic: p < 0.003; S1, Ile -> Val: p < 0.003; S2, Gly --> Gly: p < 0.04; V-1 intronic: p < 0.002; V4, in 3' untranslated region: p < 0.007). Q-1, S1 and V-1 were also associated with ppFEV1, FEV1/FVC ratio and ppFEF25-75 (p values 0.001 - 0.02). S2 was associated with FEV1/FVC ratio (p < 0.05). The association between S1 and residual volume revealed a trend toward significance (p value < 0.07). Linkage disequilibrium and haplotype analyses suggested that S1 had the strongest degree of association with COPD and pulmonary function abnormalities. CONCLUSION: Five SNPs in ADAM33 were associated with COPD and lung function in long-term smokers. Functional studies will be needed to evaluate the biologic significance of these polymorphisms in the pathogenesis of COPD. PMID- 19284603 TI - Dissecting the bacterial type VI secretion system by a genome wide in silico analysis: what can be learned from available microbial genomic resources? AB - BACKGROUND: The availability of hundreds of bacterial genomes allowed a comparative genomic study of the Type VI Secretion System (T6SS), recently discovered as being involved in pathogenesis. By combining comparative and phylogenetic approaches using more than 500 prokaryotic genomes, we characterized the global T6SS genetic structure in terms of conservation, evolution and genomic organization. RESULTS: This genome wide analysis allowed the identification of a set of 13 proteins constituting the T6SS protein core and a set of conserved accessory proteins. 176 T6SS loci (encompassing 92 different bacteria) were identified and their comparison revealed that T6SS-encoded genes have a specific conserved genetic organization. Phylogenetic reconstruction based on the core genes showed that lateral transfer of the T6SS is probably its major way of dissemination among pathogenic and non-pathogenic bacteria. Furthermore, the sequence analysis of the VgrG proteins, proposed to be exported in a T6SS dependent way, confirmed that some C-terminal regions possess domains showing similarities with adhesins or proteins with enzymatic functions. CONCLUSION: The core of T6SS is composed of 13 proteins, conserved in both pathogenic and non pathogenic bacteria. Subclasses of T6SS differ in regulatory and accessory protein content suggesting that T6SS has evolved to adapt to various microenvironments and specialized functions. Based on these results, new functional hypotheses concerning the assembly and function of T6SS proteins are proposed. PMID- 19284604 TI - Assessment of human resources for health using cross-national comparison of facility surveys in six countries. AB - BACKGROUND: Health facility assessments are being increasingly used to measure and monitor indicators of health workforce performance, but the global evidence base remains weak. Partly this is due to the wide variability in assessment methods and tools, hampering comparability across and within countries and over time. The World Health Organization coordinated a series of facility-based surveys using a common approach in six countries: Chad, Cote d'Ivoire, Jamaica, Mozambique, Sri Lanka and Zimbabwe. The objectives were twofold: to inform the development and monitoring of human resources for health (HRH) policy within the countries; and to test and validate the use of standardized facility-based human resources assessment tools across different contexts. METHODS: The survey methodology drew on harmonized questionnaires and guidelines for data collection and processing. In accordance with the survey's dual objectives, this paper presents both descriptive statistics on a number of policy-relevant indicators for monitoring and evaluation of HRH as well as a qualitative assessment of the usefulness of the data collection tool for comparative analyses. RESULTS: The findings revealed a large diversity in both the organization of health services delivery and, in particular, the distribution and activities of facility-based health workers across the sampled countries. At the same time, some commonalities were observed, including the importance of nursing and midwifery personnel in the skill mix and the greater tendency of physicians to engage in dual practice. While the use of standardized questionnaires offered the advantage of enhancing cross-national comparability of the results, some limitations were noted, especially in relation to the categories used for occupations and qualifications that did not necessarily conform to the country situation. CONCLUSION: With increasing experience in health facility assessments for HRH monitoring comes greater need to establish and promote best practices regarding methods and tools for their implementation, as well as dissemination and use of the results for evidence-informed decision-making. The overall findings of multi-country facility based survey should help countries and partners develop greater capacity to identify and measure indicators of HRH performance via this approach, and eventually contribute to better understanding of health workforce dynamics at the national and international levels. PMID- 19284606 TI - The EQ-5D (Euroqol) is a valid generic instrument for measuring quality of life in patients with dyspepsia. AB - BACKGROUND: There is little information of the validity of generic instruments in measuring health-related quality of life (HRQOL) in patients with dyspepsia. We aimed to assess the reliability and validity of the EQ-5D, a brief and simple instrument, in measuring HRQOL in adult patients with dyspepsia. METHODS: Consecutive adults with dyspepsia attending the Gastroenterology clinic in a tertiary referral center were interviewed with the EQ-5D (both English and Malay versions), the short-form Nepean Dyspepsia Index (SF-NDI), the SF-36 and Leeds Dyspepsia Questionnaire (LDQ). Known-groups and convergent construct validity were investigated by testing hypotheses at attribute and overall levels. A repeat telephone interview was conducted 2 weeks later to assess test-retest reliability. RESULTS: A total of 113 patients (mean (SD) age: 53.7 (14) years; 49.5% male; 24.8% Malays, 37.2% Chinese; 70.8% functional dyspepsia) were recruited. Response rate was 100% with nil missing data. Known-groups validation revealed 20/26 hypotheses fulfillment. Patients with more severe dyspepsia reported more problems with their usual activity (p = 0.07) and pain (p = 0.06) and demonstrated lower median VAS scores (60 vs 70, p = 0.002) and EQ-5D utility scores (0.72 vs 0.78, p = 0.002). Those reporting problems in various EQ-5D dimensions had significantly lower scores in relevant SF-36 and SF-NDI dimensions. The overall EQ-5D utility score also demonstrated good correlation with the SF-36 summary physical and mental scores and the SF-NDI total score. Intraclass correlation coefficient for test-retest reliability was 0.66 (95% CI = 0.55 - 0.76). CONCLUSION: The EQ-5D is an acceptable, valid and reliable generic instrument for measuring HRQOL in adult patients with dyspepsia. PMID- 19284605 TI - Regression of orthotopic neuroblastoma in mice by targeting the endothelial and tumor cell compartments. AB - BACKGROUND: High-risk neuroblastoma has an overall five-year survival of less than 40%, indicating a need for new treatment strategies such as angiogenesis inhibition. Recent studies have shown that chemotherapeutic drugs can inhibit angiogenesis if administered in a continuous schedule. The aim of this study was primarily to characterize tumor spread in an orthotopic, metastatic model for aggressive, MYCN-amplified neuroblastoma and secondarily to study the effects of daily administration of the chemotherapeutic agent CHS 828 on tumor angiogenesis, tumor growth, and spread. METHODS: MYCN-amplified human neuroblastoma cells (IMR 32, 2 x 10(6)) were injected into the left adrenal gland in SCID mice through a flank incision. Nine weeks later, a new laparotomy was performed to confirm tumor establishment and to estimate tumor volume. Animals were randomized to either treatment with CHS 828 (20 mg/kg/day; p.o.) or vehicle control. Differences between groups in tumor volume were analyzed by Mann-Whitney U test and in metastatic spread using Fisher's exact test. Differences with p < 0.05 were considered statistically significant. RESULTS: The orthotopic model resembled clinical neuroblastoma in respect to tumor site, growth and spread. Treatment with CHS 828 resulted in tumor regression (p < 0.001) and reduction in viable tumor fraction (p < 0.001) and metastatic spread (p < 0.05) in correlation with reduced plasma levels of the putative tumor marker chromogranin A (p < 0.001). These effects were due to increased tumor cell death and reduced angiogenesis. No treatment-related toxicities were observed. CONCLUSION: The metastatic animal model in this study resembled clinical neuroblastoma and is therefore clinically relevant for examining new treatment strategies for this malignancy. Our results indicate that daily scheduling of CHS 828 may be beneficial in treating patients with high-risk neuroblastoma. PMID- 19284607 TI - Prevalence of pathogenetic MC4R mutations in Italian children with early onset obesity, tall stature and familial history of obesity. AB - BACKGROUND: Melanocortin-4-receptor (MC4R) mutations represent the most frequent genetic cause of non-syndromic early onset obesity. Children carrying MC4R mutations seem to show a particular phenotype characterized by early onset, severe obesity and high stature. To verify whether MC4R mutations are associated with this particular phenotype in the Italian pediatric population, we decided to screen the MC4R gene in a group of obese children selected on the basis of their phenotype. METHODS: To perform this study, a multicentric approach was designed. Particularly, to be enrolled in the study subjects needed to meet the following criteria: Body mass index > or = 3 deviation scores according to age and sex, familiar history of obesity (at least one parent obese), obesity onset before the 10 years old, height > or = 2 deviation scores. The coding region of MC4R gene was screened in 240 obese children (mean age 8.3 +/- 3.1, mean BMI 30.8 +/- 5.4) and in 200 controls (mean age 8.1 +/- 2.8; mean BMI 14.2 +/- 2.5). RESULTS: Three mutations have been found in five obese children. The S127L (C380T), found in three unrelated children, had been described and functionally characterized previously. The Q307X (C919T) and the Y332H (T994C) mutations were found in two patients. Functional studies showed that only Q307X impaired protein function. CONCLUSION: The low prevalence of MC4R mutations (1.6%) in this group of obese children selected according to the obesity degree, the tall stature and the family history of obesity was similar to the prevalence observed in previous screenings performed in obese adults and in not phenotypically selected obese children. PMID- 19284608 TI - An unusual presentation of precursor T cell lymphoblastic leukemia/lymphoma with cholestatic jaundice: case report. AB - BACKGROUND: Cholestatic jaundice as a presenting symptom of Precursor T lymphoblastic leukemia (T-ALL)/lymphoma (T-LBL) has never been reported in literature. Similarly, precursor T-ALL/T-LBL is characteristically negative for synaptophysin. We report the first case of a patient with precursor T-ALL/T-LBL who presented with cholestatic jaundice and aberrant tumor expression of synaptophysin. CASE REPORT: 42 year old male presented with anorexia, nausea, jaundice, pale stools, dark urine and about 35 pound weight loss over the previous 3 weeks. The initial laboratory work was suggestive of cholestatic jaundice. Markedly elevated LDH (2025 U/L) and CA 19-9 (1778 u/ML) were also noticed. The CT scan of abdomen showed massive hepatomegaly with coarse echotexture with contracted gall bladder and normal sized common bile duct. Chest x-ray revealed a mediastinal mass with mediastinal widening. CT scan of the chest showed anterior mediastinal mass (16 cm x 10 cm). CT guided biopsy of the mass showed malignant lymphoma with diffuse proliferation of medium sized lymphoid cells. The neoplastic cells were positive for CD1a, CD3, CD4, CD5, CD8 and CD43 with aberrant expression of synaptophysin. PET CT scan again showed a large anterior mediastinal mass with diffuse liver involvement and abnormal activity in axial bones. CT guided liver biopsy and bone marrow biopsy revealed the same morphology and immunohistochemistry. Bone marrow aspirate showed 85% lymphoblasts. Thus, the diagnosis of precursor T-ALL/T-LBL was made and jaundice with elevated CA 19-9 were attributed to intrahepatic cholestasis. CONCLUSION: Our case illustrates an unusual presentation of hematological malignancies as cholestatic jaundice. It also indicates the non-specific nature of CA 19-9 for pancreaticobiliary malignancies. It is the first case report of neoplastic precursor T cell lymphoblasts with unusual expression of synaptophysin. Tissue biopsy with thorough immunohistochemistry is required to differentiate precursor T-ALL/T-LBL from thymoma and small cell carcinoma. PMID- 19284609 TI - On the significance of Surfactant Protein-A within the human lungs. AB - Surfactant Protein-A (SP-A) is the most prominent among four proteins in the pulmonary surfactant-system. SP-A is expressed by alveolar epithelial cells type II as well as by a portion of non small cell lung carcinomas (NSCLC).The expression of SP-A is complexly regulated on the transcriptional and the chromosomal level. SP-A is a major player in the pulmonary cytokine-network and moreover has been described to act in the pulmonary host defense.By the use of cell culture or animal models the functional properties have been repeatedly shown in many aspects, often bearing surprising properties which strongly indicate the physiological importance of SP-A. To date SP-A is recognized as a molecule essential for pulmonary development, structure and function. An upcoming number of reports deals with the role of SP-A for pulmonary pathology. This article gives an overview about the state of knowledge on SP-A focused in applications for human pulmonary disorders and points out the importance for pathology-orientated research approaches using immunohistochemistry or in situ hybridization as promising methods to further elucidate the role of this molecule in adult lung diseases. PMID- 19284611 TI - A randomised control trial of the effectiveness of personalised letters sent subsequent to school dental inspections in increasing registration in unregistered children. AB - BACKGROUND: Recent studies have cast doubt on the effectiveness and efficiency of school based dental screening programmes in improving dental attendance or improving dental health. In 2002 the National Dental Inspection Programme was introduced in Scotland which categorises children by their dental health and informs parents of the findings via a personalised letter home and encourages dental registration. In addition, epidemiological data for local and national planning purposes is collected. This replaced an earlier school screening system in Lothian where a generic letter urging registration was sent to children who were identified as not being registered with a dentist. The objective of this study is to compare dental registrations rates among unregistered children in these two school inspection systems with a system where letters were sent home but no dental inspection was carried out. METHODS: The study was designed as a single blinded, cluster randomised, controlled trial involving 12,765 12-13-year old children attending all 65 state Secondary schools in Lothian and Fife during the academic year 2003/4.After stratifying for school size and range of social deprivation, schools were randomly allocated to one of four groups:1. 'Traditional' inspection, letter to unregistered children only,2. Letter sent home to unregistered children only, no inspection,3. National Dental Inspection Programme, letter to all children,4. Control group in which the children were neither inspected nor sent a letter.Dental Registration status was compared at baseline and 3 months post inspection. RESULTS: The registration levels in both the 'Traditional' screening and the NDIP inspection groups rose 3 months post inspection (14% and 15.8% respectively) but were not significantly different from one another or the control group which rose by 15.8% (p > 0.05). The group who were sent a letter home but were not inspected also has a rise in registration levels of 18.1% which was not significantly different from either of the groups who were inspected or the control group (p > 0.05). The only significant predictors of registration were previous registration (p < 0.05) and within those who previously registered, the length of time since last registration (P < 0.001). CONCLUSION: Neither of the two dental inspection methods nor a letter home to unregistered children resulted in a significant rise in registration rates in 12-13-year-olds compared to a control group of children who received no intervention. PMID- 19284610 TI - Connexins: a myriad of functions extending beyond assembly of gap junction channels. AB - Connexins constitute a large family of trans-membrane proteins that allow intercellular communication and the transfer of ions and small signaling molecules between cells. Recent studies have revealed complex translational and post-translational mechanisms that regulate connexin synthesis, maturation, membrane transport and degradation that in turn modulate gap junction intercellular communication. With the growing myriad of connexin interacting proteins, including cytoskeletal elements, junctional proteins, and enzymes, gap junctions are now perceived, not only as channels between neighboring cells, but as signaling complexes that regulate cell function and transformation. Connexins have also been shown to form functional hemichannels and have roles altogether independent of channel functions, where they exert their effects on proliferation and other aspects of life and death of the cell through mostly-undefined mechanisms. This review provides an updated overview of current knowledge of connexins and their interacting proteins, and it describes connexin modulation in disease and tumorigenesis. PMID- 19284612 TI - Cardiovascular magnetic resonance in the diagnosis of acute heart transplant rejection: a review. AB - BACKGROUND: Screening for organ rejection is a critical component of care for patients who have undergone heart transplantation. Endomyocardial biopsy is the gold standard screening tool, but non-invasive alternatives are needed. Cardiovascular magnetic resonance (CMR) is well suited to provide an alternative to biopsy because of its ability to quantify ventricular function, morphology, and characterize myocardial tissue. CMR is not widely used to screen for heart transplant rejection, despite many trials supporting its use for this indication. This review summarizes the different CMR sequences that can detect heart transplant rejection as well as the strengths and weaknesses of their application. RESULTS: T2 quantification by spin echo techniques has been criticized for poor reproducibility, but multiple studies show its utility in screening for rejection. Human and animal data estimate that T2 quantification can diagnose rejection with sensitivities and specificities near 90%. There is also a suggestion that T2 quantification can predict rejection episodes in patients with normal endomyocardial biopsies.T1 quantification has also shown association with biopsy proven rejection in a small number of trials. T1 weighted gadolinium early enhancement appeared promising in animal data, but has had conflicting results in human trials. Late gadolinium enhancement in the diagnosis of rejection has not been evaluated.CMR derived measures of ventricular morphology and systolic function have insufficient sensitivity to diagnose mild to moderate rejection. CMR derived diastolic function can demonstrate abnormalities in allografts compared to native human hearts, but its ability to diagnose rejection has not yet been tested.There is promising animal data on the ability of iron oxide contrast agents to illustrate the changes in vascular permeability and macrophage accumulation seen in rejection. Despite good safety data, these contrast agents have not been tested in the human heart transplant population. CONCLUSION: T2 quantification has demonstrated the best correlation to biopsy proven heart transplant rejection. Further studies evaluating diastolic function, late gadolinium enhancement, and iron oxide contrast agents to diagnose rejection are needed. Future studies should focus on combining multiple CMR measures into a transplant rejection scoring system which would improve sensitivity and possibly reduce, if not eliminate, the need for endomyocardial biopsy. PMID- 19284613 TI - A national cross-sectional study on nurses' intent to leave and job satisfaction in Lebanon: implications for policy and practice. AB - BACKGROUND: Lebanon is perceived to be suffering from excessive nurse migration, low job satisfaction, poor retention and high turnover. Little is known about the magnitude of nurse migration and predictors of intent to leave. The objective of this study is to determine the extent of nurses' intent to leave and examine the impact of job satisfaction on intent to leave. Intent to leave was explored to differentiate between nurses who intend to leave their current hospital and those intending to leave the country. METHODS: A cross-sectional design was used to survey nurses currently practicing in Lebanese hospitals. A total of 1,793 nurses employed in 69 hospitals were surveyed. Questions included those relating to demographic characteristics, intent to leave, and the McCloskey Mueller Satisfaction Scale. Univariate descriptive statistics were conducted on sample's demographic characteristics including gender, age, marital status and educational level. Bivariate associations between intent to leave and demographic characteristics were tested using Pearson Chi-square. Differences in satisfaction scores between nurses with and without intent to leave were tested using t-test and ANOVA f-test. A multinomial logistic regression model was created to predict intent to leave the hospital and intent to leave the country. RESULTS: An alarming 67.5% reported intent to leave within the next 1 to 3 years, many of whom disclosed intent to leave the country (36.7%). Within nurses who reported an intent to leave the hospital but stay in Lebanon, 22.1% plan to move to a different health organization in Lebanon, 29.4% plan to leave the profession and 48.5% had other plans. Nurses reported being least satisfied with extrinsic rewards. A common predictor of intent to leave the hospital and the country was dissatisfaction with extrinsic rewards. Other predictors of intent to leave (country or hospital) included age, gender, marital status, degree type, and dissatisfaction with scheduling, interaction opportunities, and control and responsibility. CONCLUSION: Study findings demonstrate linkages between job satisfaction, intent to leave, and migration in a country suffering from a nursing shortage. Findings can be used by health care managers and policy makers in managing job satisfaction, intent to leave and nurse migration. PMID- 19284614 TI - The Neuroscience Peer Review Consortium. AB - As the Neuroscience Peer Review Consortium (NPRC) ends its first year, it is worth looking back to see how the experiment has worked. In order to encourage dissemination of the details outlined in this Editorial, it will also be published in other journals in the Neuroscience Peer Review Consortium. PMID- 19284615 TI - Identification of subtelomeric genomic imbalances and breakpoint mapping with quantitative PCR in 296 individuals with congenital defects and/or mental retardation. AB - BACKGROUND: Submicroscopic imbalances in the subtelomeric regions of the chromosomes are considered to play an important role in the aetiology of mental retardation (MR). The aim of the study was to evaluate a quantitative PCR (qPCR) protocol established by Boehm et al. (2004) in the clinical routine of subtelomeric testing. RESULTS: 296 patients with MR and a normal karyotype (500 550 bands) were screened for subtelomeric imbalances by using qPCR combined with SYBR green detection. In total, 17 patients (5.8%) with 20 subtelomeric imbalances were identified. Six of the aberrations (2%) were classified as causative for the symptoms, because they occurred either de novo in the patients (5 cases) or the aberration were be detected in the patient and an equally affected parent (1 case). The extent of the deletions ranged from 1.8 to approximately 10 Mb, duplications were 1.8 to approximately 5 Mb in size. In 6 patients, the copy number variations (CNVs) were rated as benign polymorphisms, and the clinical relevance of these CNVs remains unclear in 5 patients (1.7%). Therefore, the overall frequency of clinically relevant imbalances ranges between 2% and 3.7% in our cohort. CONCLUSION: This study illustrates that the qPCR/SYBR green technique represents a rapid and versatile method for the detection of subtelomeric imbalances and the option to map the breakpoint. Thus, this technique is highly suitable for genotype/phenotype studies in patients with MR/developmental delay and/or congenital defects. PMID- 19284616 TI - Genetic basis of arsenite and cadmium tolerance in Saccharomyces cerevisiae. AB - BACKGROUND: Arsenic and cadmium are widely distributed in nature and pose serious threats to the environment and human health. Exposure to these nonessential toxic metals may result in a variety of human diseases including cancer. However, arsenic and cadmium toxicity targets and the cellular systems contributing to tolerance acquisition are not fully known. RESULTS: To gain insight into metal action and cellular tolerance mechanisms, we carried out genome-wide screening of the Saccharomyces cerevisiae haploid and homozygous diploid deletion mutant collections and scored for reduced growth in the presence of arsenite or cadmium. Processes found to be required for tolerance to both metals included sulphur and glutathione biosynthesis, environmental sensing, mRNA synthesis and transcription, and vacuolar/endosomal transport and sorting. We also identified metal-specific defence processes. Arsenite-specific defence functions were related to cell cycle regulation, lipid and fatty acid metabolism, mitochondrial biogenesis, and the cytoskeleton whereas cadmium-specific defence functions were mainly related to sugar/carbohydrate metabolism, and metal-ion homeostasis and transport. Molecular evidence indicated that the cytoskeleton is targeted by arsenite and that phosphorylation of the Snf1p kinase is required for cadmium tolerance. CONCLUSION: This study has pin-pointed core functions that protect cells from arsenite and cadmium toxicity. It also emphasizes the existence of both common and specific defence systems. Since many of the yeast genes that confer tolerance to these agents have homologues in humans, similar biological processes may act in yeast and humans to prevent metal toxicity and carcinogenesis. PMID- 19284617 TI - The correlation and level of agreement between end-tidal and blood gas pCO2 in children with respiratory distress: a retrospective analysis. AB - BACKGROUND: To investigate the correlation and level of agreement between end tidal carbon dioxide (EtCO2) and blood gas pCO2 in non-intubated children with moderate to severe respiratory distress. METHODS: Retrospective study of patients admitted to an intermediate care unit (InCU) at a tertiary care center over a 20 month period with moderate to severe respiratory distress secondary to asthma, bronchiolitis, or pneumonia. Patients with venous pCO2 (vpCO2) and EtCO2 measurements within 10 minutes of each other were eligible for inclusion. Patients with cardiac disease, chronic pulmonary disease, poor tissue perfusion, or metabolic abnormalities were excluded. RESULTS: Eighty EtCO2-vpCO2 paired values were available from 62 patients. The mean +/- SD for EtCO2 and vpCO2 was 35.7 +/- 10.1 mmHg and 39.4 +/- 10.9 mmHg respectively. EtCO2 and vpCO2 values were highly correlated (r = 0.90, p < 0.0001). The correlations for asthma, bronchiolitis and pneumonia were 0.74 (p < 0.0001), 0.83 (p = 0.0002) and 0.98 (p < 0.0001) respectively. The mean bias +/- SD between EtCO2 and vpCO2 was -3.68 +/ 4.70 mmHg. The 95% level of agreement ranged from -12.88 to +5.53 mmHg. EtCO2 was found to be more accurate when vpCO2 was 35 mmHg or lower. CONCLUSION: EtCO2 is correlated highly with vpCO2 in non-intubated pediatric patients with moderate to severe respiratory distress across respiratory illnesses. Although the level of agreement between the two methods precludes the overall replacement of blood gas evaluation, EtCO2 monitoring remains a useful, continuous, non-invasive measure in the management of non-intubated children with moderate to severe respiratory distress. PMID- 19284618 TI - Reconstruction of metabolic pathways for the cattle genome. AB - BACKGROUND: Metabolic reconstruction of microbial, plant and animal genomes is a necessary step toward understanding the evolutionary origins of metabolism and species-specific adaptive traits. The aims of this study were to reconstruct conserved metabolic pathways in the cattle genome and to identify metabolic pathways with missing genes and proteins. The MetaCyc database and PathwayTools software suite were chosen for this work because they are widely used and easy to implement. RESULTS: An amalgamated cattle genome database was created using the NCBI and Ensembl cattle genome databases (based on build 3.1) as data sources. PathwayTools was used to create a cattle-specific pathway genome database, which was followed by comprehensive manual curation for the reconstruction of metabolic pathways. The curated database, CattleCyc 1.0, consists of 217 metabolic pathways. A total of 64 mammalian-specific metabolic pathways were modified from the reference pathways in MetaCyc, and two pathways previously identified but missing from MetaCyc were added. Comparative analysis of metabolic pathways revealed the absence of mammalian genes for 22 metabolic enzymes whose activity was reported in the literature. We also identified six human metabolic protein coding genes for which the cattle ortholog is missing from the sequence assembly. CONCLUSION: CattleCyc is a powerful tool for understanding the biology of ruminants and other cetartiodactyl species. In addition, the approach used to develop CattleCyc provides a framework for the metabolic reconstruction of other newly sequenced mammalian genomes. It is clear that metabolic pathway analysis strongly reflects the quality of the underlying genome annotations. Thus, having well-annotated genomes from many mammalian species hosted in BioCyc will facilitate the comparative analysis of metabolic pathways among different species and a systems approach to comparative physiology. PMID- 19284619 TI - A family of splice variants of CstF-64 expressed in vertebrate nervous systems. AB - BACKGROUND: Alternative splicing and polyadenylation are important mechanisms for creating the proteomic diversity necessary for the nervous system to fulfill its specialized functions. The contribution of alternative splicing to proteomic diversity in the nervous system has been well documented, whereas the role of alternative polyadenylation in this process is less well understood. Since the CstF-64 polyadenylation protein is known to be an important regulator of tissue specific polyadenylation, we examined its expression in brain and other organs. RESULTS: We discovered several closely related splice variants of CstF-64 - collectively called betaCstF-64 - that could potentially contribute to proteomic diversity in the nervous system. The betaCstF-64 splice variants are found predominantly in the brains of several vertebrate species including mice and humans. The major betaCstF-64 variant mRNA is generated by inclusion of two alternate exons (that we call exons 8.1 and 8.2) found between exons 8 and 9 of the CstF-64 gene, and contains an additional 147 nucleotides, encoding 49 additional amino acids. Some variants of betaCstF-64 contain only the first alternate exon (exon 8.1) while other variants contain both alternate exons (8.1 and 8.2). In mice, the predominant form of betaCstF-64 also contains a deletion of 78 nucleotides from exon 9, although that variant is not seen in any other species examined, including rats. Immunoblot and 2D-PAGE analyses of mouse nuclear extracts indicate that a protein corresponding to betaCstF-64 is expressed in brain at approximately equal levels to CstF-64. Since betaCstF-64 splice variant family members were found in the brains of all vertebrate species examined (including turtles and fish), this suggests that betaCstF-64 has an evolutionarily conserved function in these animals. betaCstF-64 was present in both pre- and post-natal mice and in different regions of the nervous system, suggesting an important role for betaCstF-64 in neural gene expression throughout development. Finally, experiments in representative cell lines suggest that betaCstF-64 is expressed in neurons but not glia. CONCLUSION: This is the first report of a family of splice variants encoding a key polyadenylation protein that is expressed in a nervous system-specific manner. We propose that betaCstF-64 contributes to proteomic diversity by regulating alternative polyadenylation of neural mRNAs. PMID- 19284620 TI - Effects of dietary Na+ deprivation on epithelial Na+ channel (ENaC), BDNF, and TrkB mRNA expression in the rat tongue. AB - BACKGROUND: In rodents, dietary Na+ deprivation reduces gustatory responses of primary taste fibers and central taste neurons to lingual Na+ stimulation. However, in the rat taste bud cells Na+ deprivation increases the number of amiloride sensitive epithelial Na+ channels (ENaC), which are considered as the "receptor" of the Na+ component of salt taste. To explore the mechanisms, the expression of the three ENaC subunits (alpha, beta and gamma) in taste buds were observed from rats fed with diets containing either 0.03% (Na+ deprivation) or 1% (control) NaCl for 15 days, by using in situ hybridization and real-time quantitative RT-PCR (qRT-PCR). Since BDNF/TrkB signaling is involved in the neural innervation of taste buds, the effects of Na+ deprivation on BDNF and its receptor TrkB expression in the rat taste buds were also examined. RESULTS: In situ hybridization analysis showed that all three ENaC subunit mRNAs were found in the rat fungiform taste buds and lingual epithelia, but in the vallate and foliate taste buds, only alpha ENaC mRNA was easily detected, while beta and gamma ENaC mRNAs were much less than those in the fungiform taste buds. Between control and low Na+ fed animals, the numbers of taste bud cells expressing alpha, beta and gamma ENaC subunits were not significantly different in the fungiform, vallate and foliate taste buds, respectively. Similarly, qRT-PCR also indicated that Na+ deprivation had no effect on any ENaC subunit expression in the three types of taste buds. However, Na+ deprivation reduced BDNF mRNA expression by 50% in the fungiform taste buds, but not in the vallate and foliate taste buds. The expression of TrkB was not different between control and Na+ deprived rats, irrespective of the taste papillae type. CONCLUSION: The findings demonstrate that dietary Na+ deprivation does not change ENaC mRNA expression in rat taste buds, but reduces BDNF mRNA expression in the fungiform taste buds. Given the roles of BDNF in survival of cells and target innervation, our results suggest that dietary Na+ deprivation might lead to a loss of gustatory innervation in the mouse fungiform taste buds. PMID- 19284621 TI - Regulation of callose synthase activity in situ in alamethicin-permeabilized Arabidopsis and tobacco suspension cells. AB - BACKGROUND: The cell wall component callose is mainly synthesized at certain developmental stages and after wounding or pathogen attack. Callose synthases are membrane-bound enzymes that have been relatively well characterized in vitro using isolated membrane fractions or purified enzyme. However, little is known about their functional properties in situ, under conditions when the cell wall is intact. To allow in situ investigations of the regulation of callose synthesis, cell suspensions of Arabidopsis thaliana (Col-0), and tobacco (BY-2), were permeabilized with the channel-forming peptide alamethicin. RESULTS: Nucleic acid binding dyes and marker enzymes demonstrated alamethicin permeabilization of plasma membrane, mitochondria and plastids, also allowing callose synthase measurements. In the presence of alamethicin, Ca2+ addition was required for callose synthase activity, and the activity was further stimulated by Mg2+ Cells pretreated with oryzalin to destabilize the microtubules prior to alamethicin permeabilization showed significantly lower callose synthase activity as compared to non-treated cells. As judged by aniline blue staining, the callose formed was deposited both at the cell walls joining adjacent cells and at discrete punctate locations earlier described as half plasmodesmata on the outer walls. This pattern was unaffected by oryzalin pretreatment, showing a quantitative rather than a qualitative effect of polymerized tubulin on callose synthase activity. No callose was deposited unless alamethicin, Ca2+ and UDP-glucose were present. Tubulin and callose synthase were furthermore part of the same plasma membrane protein complex, as judged by two-dimensional blue native SDS-PAGE. CONCLUSION: Alamethicin permeabilization allowed determination of callose synthase regulation and tubulin interaction in the natural crowded cellular environment and under conditions where contacts between the cell wall, the plasma membrane and cytoskeletal macromolecules remained. The results also suggest that alamethicin permeabilization induces a defense response mimicking the natural physical separation of cells (for example when intercellulars are formed), during which plasmodesmata are transiently left open. PMID- 19284622 TI - Conventional intramuscular sedatives versus ziprasidone for severe agitation in adolescents: case-control study. AB - OBJECTIVE: The objective of this study was to compare intramuscular (IM) ziprasidone to conventional IM medications (haloperidol combined with lorazepam) for the treatment of severe agitation in adolescents (age 12-17). METHODS: We retrospectively identified consecutive severe agitation episodes (defined as requiring physical restraint) in adolescents treated with either IM ziprasidone or conventional IM agents in a psychiatric emergency room. For ziprasidone, the dosage was 20 mg for 23 episodes and 10 mg for 5 episodes. For 24 episodes treated with combined haloperidol and lorazepam, the dosages were 4.8 +/- 0.3 SEM mg and 1.9 +/- 0.4 mg respectively. Outcomes were the duration of restraint and need for adjunctive "rescue" medications within 60 minutes. These outcomes were decided prior to reviewing any records. RESULTS: No difference was found in restraint duration (ziprasidone, N = 28, 55 +/- 5 minutes; haloperidol with lorazepam N = 24, 65 +/- 7 minutes, P = NS). Use of "rescue" medications did not differ between the two groups. No changes in blood pressure were found, but pulse decreased 8.3 +/- 2.4 for haloperidol with lorazepam and 8.9 +/- 4.24 for ziprasidone (P = NS). No instances of excessive sedation or extra-pyramidal symptoms were documented. CONCLUSION: In this study, IM ziprasidone appeared effective, well tolerated, and similar in clinical profile to combined conventional IM medications for treating severe agitation in adolescents. Given the reportedly favorable acute side effect profile of parenteral atypical agents, they may provide an alternative to conventional antipsychotics for treating acute agitation in both adult and adolescent populations. Future randomized, controlled studies are needed. PMID- 19284623 TI - Sunitinib treatment for patients with clear-cell metastatic renal cell carcinoma: clinical outcomes and plasma angiogenesis markers. AB - BACKGROUND: Sunitinib is a protein tyrosine kinase-inhibitor targeting VEGFR, c kit and PDGFR. It has been approved for the treatment of metastatic renal-cell carcinoma and gastrointestinal stromal tumors. Although it has been shown to prolong disease-free and overall survival in renal-cell carcinoma patients, only 70% of the treated population receive a clinical benefit (CB) from the treatment. Markers that could predict clinical benefit to sunitinib would be an important aid in monitoring and following their treatment. We assessed the outcome and plasma proangiogenic factors in patients with metastatic renal cell carcinoma (mRCC) treated with sunitinib in our institution. METHODS: We have treated 42 patients with metastatic clear-cell renal carcinoma with sunitinib. Plasma concentrations of VEGF-A, sVEGFR2 and PDGF were determined by ELISA. RESULTS: At the time of analysis 39 patients were evaluable for response and 30 patients had obtained a clinical benefit (CB). Median progression-free survival was 268 days (8.93 months) and median overall survival was 487 days (16.23 months). Interestingly, disease stabilization or objective response resulted in comparable overall survival. Most treatment-related adverse events were of mild-to-moderate intensity with one treatment-related death. Plasma sVEGFR2 and PDGF levels had no predictive value. Fold-increase in plasma VEGF was significantly lower in patients that obtained a CB as compared to patients that progressed after two cycles of treatment. Plasma VEGF did not increase in patients with initial CB at the time of progression. CONCLUSION: Sunitinib showed substantial activity in mRCC. Disease stabilization or objective response resulted in comparable overall survival and both outcomes should be considered positive. Fold-increase in plasma VEGF predicts for CB and could be a candidate marker. Progression after initial CB is not associated with elevated plasma VEGF, implying a different mechanism of resistance. PMID- 19284624 TI - Particles induce apical plasma membrane enlargement in epithelial lung cell line depending on particle surface area dose. AB - BACKGROUND: Airborne particles entering the respiratory tract may interact with the apical plasma membrane (APM) of epithelial cells and enter them. Differences in the entering mechanisms of fine (between 0.1 microm and 2.5 microm) and ultrafine ( or= 70) for all nodes were reached in all datasets, but an increase in data did not necessarily equate to an increase in support values. However, simulations based on a single empirical locus reached higher overall levels of support with less data than did the simulations that were based on all seven empirical nuclear loci, and symmetric tree distances were much lower for single versus multiple gene simulation analyses. CONCLUSION: Our empirical results provide new insights into the phylogenetics of the Emydidae, but the short branches recovered deep in the tree also indicate the need for additional work on this clade to recover all intergeneric relationships with confidence and to delimit species for some problematic groups. Our simulation results suggest that moderate (in the few-to-tens of kb range) amounts of data are necessary to recover most emydid relationships with high support values. They also suggest that previous simulations that do not incorporate among-gene tree topological variance probably underestimate the amount of data needed to recover well supported phylogenies. PMID- 19284631 TI - Development and preliminary validation of a Family Nutrition and Physical Activity (FNPA) screening tool. AB - BACKGROUND: Parents directly influence children's physical activity and nutrition behaviors and also dictate the physical and social environments that are available to their children. This paper summarizes the development of an easy to use screening tool (The Family Nutrition and Physical Activity (FNPA) Screening Tool) designed to assess family environmental and behavioral factors that may predispose a child to becoming overweight. METHODS: The FNPA instrument was developed using constructs identified in a comprehensive evidence analysis conducted in collaboration with the American Dietetics Association. Two or three items were created for each of the ten constructs with evidence grades of II or higher. Parents of first grade students from a large urban school district (39 schools) were recruited to complete the FNPA screening tool and provide permission to link results to BMI data obtained from trained nurses in each school. A total of 1085 surveys were completed out of the available sample of 2189 children in the district. Factor analysis was conducted to examine the factor structure of the scale. Mixed model analyses were conducted on the composite FNPA score to determine if patterns in home environments and behaviors matched some of the expected socio-economic (SES) and ethnic patterns in BMI. Correlations among FNPA constructs and other main variables were computed to examine possible associations among the various factors. Finally, logistic regression was used to evaluate the construct validity of the FNPA scale. RESULTS: Factor analyses revealed the presence of a single factor and this unidimensional structure was supported by the correlation analyses. The correlations among constructs were consistently positive but the total score had higher correlations with child BMI than the other individual constructs. The FNPA scores followed expected demographic patterns with low income families reporting lower (less favorable) scores than moderate or high income families. Children with a total score in the lowest tertile (high risk family environment and behaviors) had an odds ratio (OR) of 1.7 (95% CI = 1.07 - 2.80) compared to children with a total score in the highest tertile (more favorable family environment and behaviors) but this effect was reduced when parent BMI was included as a covariate. CONCLUSION: The results support the contention that the FNPA tool captures important elements of the family environment and behaviors that relate to risk for child overweight. PMID- 19284632 TI - Selective deficit of second language: a case study of a brain-damaged Arabic Hebrew bilingual patient. AB - BACKGROUND: An understanding of how two languages are represented in the human brain is best obtained from studies of bilingual patients who have sustained brain damage. The primary goal of the present study was to determine whether one or both languages of an Arabic-Hebrew bilingual individual are disrupted following brain damage. I present a case study of a bilingual patient, proficient in Arabic and Hebrew, who had sustained brain damage as a result of an intracranial hemorrhage related to herpes encephalitis. METHODS: The patient's performance on several linguistic tasks carried out in the first language (Arabic) and in the second language (Hebrew) was assessed, and his performance in the two languages was compared. RESULTS: The patient displayed somewhat different symptomatologies in the two languages. The results revealed dissociation between the two languages in terms of both the types and the magnitude of errors, pointing to aphasic symptoms in both languages, with Hebrew being the more impaired. Further analysis disclosed that this dissociation was apparently caused not by damage to his semantic system, but rather by damage at the lexical level. CONCLUSION: The results suggest that the principles governing the organization of lexical representations in the brain are not similar for the two languages. PMID- 19284633 TI - TFCat: the curated catalog of mouse and human transcription factors. AB - Unravelling regulatory programs governed by transcription factors (TFs) is fundamental to understanding biological systems. TFCat is a catalog of mouse and human TFs based on a reliable core collection of annotations obtained by expert review of the scientific literature. The collection, including proven and homology-based candidate TFs, is annotated within a function-based taxonomy and DNA-binding proteins are organized within a classification system. All data and user-feedback mechanisms are available at the TFCat portal (http://www.tfcat.ca). PMID- 19284634 TI - The susceptibility of Anopheles lesteri to infection with Korean strain of Plasmodium vivax. AB - BACKGROUND: Following its recent re-emergence, malaria has gained renewed attention as a serious infectious disease in Korea. Three species of the Hyrcanusgroup, Anopheles lesteri, Anopheles sinensis and Anopheles pullus, have long been suspected malaria vectors. However, opinions about their vector ability are controversial. The present study was designed with the aim of determining the susceptibility of these mosquitoes to a Korean isolate of Plasmodium vivax. Also, An. sinensis is primarily suspected to be vector of malaria in Korea, but in Thailand, the same species is described to have less medical importance. Therefore, comparative susceptibility of Thai and Korean strains of An. sinensis with Thai strain of P. vivax may be helpful to understand whether these geographically different strains exhibit differences in their susceptibility or not. METHODS: The comparative susceptibility of An. lesteri, An. sinensis and An. pullus was studied by feeding laboratory-reared mosquitoes on blood from patients carrying gametocytes from Korea and Thailand. RESULTS: In experimental feeding with Korean strain of P. vivax, oocysts developed in An. lesteri, An. sinensis and An. pullus. Salivary gland sporozoites were detected only in An. lesteri and An. sinensis but not in An. pullus. Large differences were found in the number of sporozoites in the salivary glands, with An. lesteri carrying much higher densities, up to 2,105 sporozoites in a single microscope field of 750 x 560 muM, whereas a maximum of 14 sporozoites were found in any individual salivary gland of An. sinensis. Similar results were obtained from a susceptibility test of two different strains of An. sinensis to Thai isolate of P. vivax, and differences in vector susceptibility according to geographical variation were not detected. CONCLUSION: The high sporozoite rate and sporozoite loads of An. lesteri indicate that this species is highly susceptible to infection with P. vivax. Anopheles sinensis appears to have a markedly reduced ability to develop salivary gland infection, whilst in An. pullus, no sporozoites were found in the salivary glands. Provided that the survival rate of An. lesteri is sufficiently high in the field, it would be a highly competent vector of vivax malaria. PMID- 19284635 TI - The experience of providing young people attending general practice with an online risk assessment tool to assess their own sexual health risk. AB - BACKGROUND: Targeted chlamydia screening has been advocated to reduce chlamydia associated reproductive sequelae. General practitioners are well positioned to play a major role in chlamydia control. The primary aim of this pilot study was to measure the effect of offering an online sexual health assessment tool, Youth Check Your Risk, on chlamydia testing rates among young people attending general practices. The secondary aim was to test the acceptability of the tool among general practitioners and young people. METHODS: General practitioners at three practices in Melbourne, Australia, referred patients aged 16 to 24 years to Youth Check Your Risk http://www.checkyourrisk.org.au for use post-consultation between March to October 2007. The proportion of young people tested for chlamydia before and during the implementation of the tool was compared. Acceptability was assessed through a structured interviewer-administered questionnaire with general practitioners, and anonymous online data provided by Youth Check Your Risk users. RESULTS: The intervention did not result in any significant increases in the proportion of 16 to 24 year old males (2.7% to 3.0%) or females (6.3% to 6.4%) tested for chlamydia. A small increase in the proportion of 16 to 19 year old females tested was seen (4.1% to 7.2%). Of the 2997 patients seen during the intervention phase, 871 (29.1%) were referred to Youth Check Your Risk and 120 used it (13.8%). Major reasons for low referral rates reported by practitioners included lack of time, discomfort with raising the issue of testing, and difficulty in remembering to refer patients. CONCLUSION: Offering an online sexual risk assessment tool in general practice did not significantly increase the proportion of young people tested for chlamydia, with GPs identifying a number of barriers to referring young people to Youth Check Your Risk. Future interventions aimed at increasing chlamydia screening in general practice with the aid of an online risk assessment tool need to identify and overcome barriers to testing. PMID- 19284636 TI - Missing call bias in high-throughput genotyping. AB - BACKGROUND: The advent of high-throughput and cost-effective genotyping platforms made genome-wide association (GWA) studies a reality. While the primary focus has been invested upon the improvement of reducing genotyping error, the problems associated with missing calls are largely overlooked. RESULTS: To probe into the effect of missing calls on GWAs, we demonstrated experimentally the prevalence and severity of the problem of missing call bias (MCB) in four genotyping technologies (Affymetrix 500 K SNP array, SNPstream, TaqMan, and Illumina Beadlab). Subsequently, we showed theoretically that MCB leads to biased conclusions in the subsequent analyses, including estimation of allele/genotype frequencies, the measurement of HWE and association tests under various modes of inheritance relationships. We showed that MCB usually leads to power loss in association tests, and such power change is greater than what could be achieved by equivalent reduction of sample size unbiasedly. We also compared the bias in allele frequency estimation and in association tests introduced by MCB with those by genotyping errors. Our results illustrated that in most cases, the bias can be greatly reduced by increasing the call-rate at the cost of genotyping error rate. CONCLUSION: The commonly used 'no-call' procedure for the observations of borderline quality should be modified. If the objective is to minimize the bias, the cut-off for call-rate and that for genotyping error rate should be properly coupled in GWA. We suggested that the ongoing QC cut-off for call-rate should be increased, while the cut-off for genotyping error rate can be reduced properly. PMID- 19284637 TI - Polymorphisms in the ADRB2 gene and Graves disease: a case-control study and a meta-analysis of available evidence. AB - BACKGROUND: The beta-2-Adrenergic receptor (ADRB2) gene on chromosome 5q33.1 is an important immunoregulatory factor. We and others have previously implicated chromosomal region 5q31-33 for contribution to the genetic susceptibility to Graves disease (GD) in East-Asian populations. Two recent studies showed associations between the single nucleotide polymorphism (SNP) rs1042714 in the ADRB2 gene and GD. In this study, we aimed to fully investigate whether the ADRB2 gene conferred susceptibility to GD in Chinese population, and to perform a meta analysis of association between ADRB2 and GD. METHODS: Approximately 1 kb upstream the transcription start site and the entire coding regions of the ADRB2 gene were resequenced in 48 Han Chinese individuals to determine the linkage disequilibrium (LD) patterns. Tag SNPs were selected and genotyped in a case control collection of 1,118 South Han Chinese subjects, which included 428 GD patients and 690 control subjects. A meta-analysis was performed with the data obtained in the present samples and those available from prior studies. RESULTS: Fifteen SNPs in the ADRB2 gene were identified by resequencing and one SNP was novel. Ten tag SNPs were investigated further to assess association of ADRB2 in the case-control collection. Neither individual tag SNP nor haplotypes showed association with GD in Han Chinese population (P > 0.05). Our meta-analysis of the ADRB2 SNP rs1042714 measured heterogeneity between the ethnic groups (I2 = 53.1%) and no association to GD was observed in the overall three studies with a random effects model (OR = 1.13, 95% CI, 0.95 to 1.36; P = 0.18). However, significant association was found from the combined data of Caucasian population with a fixed effects model (OR = 1.18, 95% CI, 1.06 to 1.32; P = 0.002; I2 = 5.9%). CONCLUSION: Our study indicated that the ADRB2 gene did not exert a substantial influence on GD susceptibility in Han Chinese population, but contributed to a detectable GD risk in Caucasian population. This inconsistency resulted largely from between-ethnicity heterogeneity. PMID- 19284638 TI - Community perceptions of mental health needs: a qualitative study in the Solomon Islands. AB - BACKGROUND: Psychosocial and mental health needs in the aftermath of conflict and disaster have attracted substantial attention. In the Solomon Islands, the conceptualisation of mental health, for several decades regarded by policy makers as primarily a health issue, has broadened and been incorporated into the national development and social policy agendas, reflecting recognition of the impact of conflict and rapid social change on the psychosocial wellbeing of the community as a whole. We sought to understand how mental health and psychosocial wellbeing were seen at the community level, the extent to which these issues were identified as being associated with periods of 'tension', violence and instability, and the availability of traditional approaches and Ministry of Health services to address these problems. METHODS: This article reports the findings of qualitative research conducted in a rural district on the island of Guadalcanal in the Solomon Islands. Key informant interviews were conducted with community leaders, and focus groups were held with women, men and young people. Wellbeing was defined broadly. RESULTS: Problems of common concern included excessive alcohol and marijuana use, interpersonal violence and abuse, teenage pregnancy, and lack of respect and cooperation. Troubled individuals and their families sought help for mental problems from various sources including chiefs, church leaders and traditional healers and, less often, trauma support workers, health clinic staff and police. Substance-related problems presented special challenges, as there were no traditional solutions at the individual or community level. Severe mental illness was also a challenge, with few aware that a community mental health service existed. Contrary to our expectations, conflict related trauma was not identified as a major problem by the community who were more concerned about the economic and social sequelae of the conflict. CONCLUSION: Communities identify and are responding to a wide range of mental health challenges; the health system generally can do more to learn about how this is being done, and build more comprehensive services and policy on this foundation. The findings underscore the need to promote awareness of those services which are available, to extend mental health care beyond urban centres to rural villages where the majority of the population live, and to promote community input to policy so as to ensure that it 'fits' the context. PMID- 19284639 TI - Influenza virus antigenic variation, host antibody production and new approach to control epidemics. AB - Influenza is an infectious disease and can lead to life-threatening complications like pneumonia. The disease is caused by three types of RNA viruses called influenza types A, B and C, each consisting of eight negative single-stranded RNA segments encoding 11 proteins. Current annual vaccines contain two type A strains and one type B strain and are capable of inducing strong antibody responses to both the surface glycoprotein hemagglutinin and the neuraminidase. While these vaccines are protective against vaccine viruses they are not effective against newly emerging viruses that contain antigenic variations known as antigenic drift and shift. In nature, environmental selection pressure generally plays a key role in selecting antigenic changes in the antigen determining spots of hemagglutinin, resulting in changes in the antigenicity of the virus. Recently, a new technology has been developed where influenza-specific IgG+ antibody-secreting plasma cells can be isolated and cloned directly from vaccinated humans and high affinity monoclonal antibodies can be produced within several weeks after vaccination. The new technology holds great promise for the development of effective passive antibody therapy to limit the spread of influenza viruses in a timely manner. PMID- 19284640 TI - Experimental exposure to diesel exhaust increases arterial stiffness in man. AB - INTRODUCTION: Exposure to air pollution is associated with increased cardiovascular morbidity, although the underlying mechanisms are unclear. Vascular dysfunction reduces arterial compliance and increases central arterial pressure and left ventricular after-load. We determined the effect of diesel exhaust exposure on arterial compliance using a validated non-invasive measure of arterial stiffness. METHODS: In a double-blind randomized fashion, 12 healthy volunteers were exposed to diesel exhaust (approximately 350 mug/m3) or filtered air for one hour during moderate exercise. Arterial stiffness was measured using applanation tonometry at the radial artery for pulse wave analysis (PWA), as well as at the femoral and carotid arteries for pulse wave velocity (PWV). PWA was performed 10, 20 and 30 min, and carotid-femoral PWV 40 min, post-exposure. Augmentation pressure (AP), augmentation index (AIx) and time to wave reflection (Tr) were calculated. RESULTS: Blood pressure, AP and AIx were generally low reflecting compliant arteries. In comparison to filtered air, diesel exhaust exposure induced an increase in AP of 2.5 mmHg (p = 0.02) and in AIx of 7.8% (p = 0.01), along with a 16 ms reduction in Tr (p = 0.03), 10 minutes post-exposure. CONCLUSION: Acute exposure to diesel exhaust is associated with an immediate and transient increase in arterial stiffness. This may, in part, explain the increased risk for cardiovascular disease associated with air pollution exposure. If our findings are confirmed in larger cohorts of susceptible populations, this simple non-invasive method of assessing arterial stiffness may become a useful technique in measuring the impact of real world exposures to combustion derived air pollution. PMID- 19284641 TI - Hyperbaric treatment for children with autism: a multicenter, randomized, double blind, controlled trial. AB - BACKGROUND: Several uncontrolled studies of hyperbaric treatment in children with autism have reported clinical improvements; however, this treatment has not been evaluated to date with a controlled study. We performed a multicenter, randomized, double-blind, controlled trial to assess the efficacy of hyperbaric treatment in children with autism. METHODS: 62 children with autism recruited from 6 centers, ages 2-7 years (mean 4.92 +/- 1.21), were randomly assigned to 40 hourly treatments of either hyperbaric treatment at 1.3 atmosphere (atm) and 24% oxygen ("treatment group", n = 33) or slightly pressurized room air at 1.03 atm and 21% oxygen ("control group", n = 29). Outcome measures included Clinical Global Impression (CGI) scale, Aberrant Behavior Checklist (ABC), and Autism Treatment Evaluation Checklist (ATEC). RESULTS: After 40 sessions, mean physician CGI scores significantly improved in the treatment group compared to controls in overall functioning (p = 0.0008), receptive language (p < 0.0001), social interaction (p = 0.0473), and eye contact (p = 0.0102); 9/30 children (30%) in the treatment group were rated as "very much improved" or "much improved" compared to 2/26 (8%) of controls (p = 0.0471); 24/30 (80%) in the treatment group improved compared to 10/26 (38%) of controls (p = 0.0024). Mean parental CGI scores significantly improved in the treatment group compared to controls in overall functioning (p = 0.0336), receptive language (p = 0.0168), and eye contact (p = 0.0322). On the ABC, significant improvements were observed in the treatment group in total score, irritability, stereotypy, hyperactivity, and speech (p < 0.03 for each), but not in the control group. In the treatment group compared to the control group, mean changes on the ABC total score and subscales were similar except a greater number of children improved in irritability (p = 0.0311). On the ATEC, sensory/cognitive awareness significantly improved (p = 0.0367) in the treatment group compared to the control group. Post-hoc analysis indicated that children over age 5 and children with lower initial autism severity had the most robust improvements. Hyperbaric treatment was safe and well tolerated. CONCLUSION: Children with autism who received hyperbaric treatment at 1.3 atm and 24% oxygen for 40 hourly sessions had significant improvements in overall functioning, receptive language, social interaction, eye contact, and sensory/cognitive awareness compared to children who received slightly pressurized room air. TRIAL REGISTRATION: clinicaltrials.gov NCT00335790. PMID- 19284642 TI - Beneficial cardiovascular effects of reducing exposure to particulate air pollution with a simple facemask. AB - BACKGROUND: Exposure to air pollution is an important risk factor for cardiovascular morbidity and mortality, and is associated with increased blood pressure, reduced heart rate variability, endothelial dysfunction and myocardial ischaemia. Our objectives were to assess the cardiovascular effects of reducing air pollution exposure by wearing a facemask. METHODS: In an open-label cross over randomised controlled trial, 15 healthy volunteers (median age 28 years) walked on a predefined city centre route in Beijing in the presence and absence of a highly efficient facemask. Personal exposure to ambient air pollution and exercise was assessed continuously using portable real-time monitors and global positional system tracking respectively. Cardiovascular effects were assessed by continuous 12-lead electrocardiographic and ambulatory blood pressure monitoring. RESULTS: Ambient exposure (PM2.5 86 +/- 61 vs 140 +/- 113 mug/m3; particle number 2.4 +/- 0.4 vs 2.3 +/- 0.4 x 104 particles/cm3), temperature (29 +/- 1 vs 28 +/- 3 degrees C) and relative humidity (63 +/- 10 vs 64 +/- 19%) were similar (P > 0.05 for all) on both study days. During the 2-hour city walk, systolic blood pressure was lower (114 +/- 10 vs 121 +/- 11 mmHg, P < 0.01) when subjects wore a facemask, although heart rate was similar (91 +/- 11 vs 88 +/- 11/min; P > 0.05). Over the 24-hour period heart rate variability increased (SDNN 65.6 +/- 11.5 vs 61.2 +/- 11.4 ms, P < 0.05; LF-power 919 +/- 352 vs 816 +/- 340 ms2, P < 0.05) when subjects wore the facemask. CONCLUSION: Wearing a facemask appears to abrogate the adverse effects of air pollution on blood pressure and heart rate variability. This simple intervention has the potential to protect susceptible individuals and prevent cardiovascular events in cities with high concentrations of ambient air pollution. PMID- 19284643 TI - The cytoplasmic 60 kDa progesterone receptor isoform predominates in the human amniochorion and placenta at term. AB - BACKGROUND: The mechanism that initiates human parturition has been proposed to be 'functional progesterone withdrawal' whereby the 116 kDa B-isoform of the progesterone receptor (PR-B) switches in favour of the 94 kDa A-isoform (PR-A) in reproductive tissues. Recently, other PR isoforms, PR-S, PR-C and PR-M generated from the same gene have been identified and partially characterised. METHODS AND RESULTS: Using immunohistochemical, western blotting and RT-PCR techniques, evidence is provided that indicates the major PR isoform present in human term fetal membranes (amnion and chorion) and syncytiotrophoblast of the placenta is neither of the classical nuclear PR-B or PR-A isoforms but is the N-terminally truncated 60 kDa PR-C isoform. Evidence is also provided that this 60 kDa isoform resides in the cytoplasm of the expressing cell types. Data are also presented to show that PR-B, PR-A and PR-S isoforms are essentially absent from the amnion and chorion, whereas PR isoforms A, B, C and S are all present in the decidua, with PR-A being the major isoform. The syncytiotrophoblast of the placenta contains the cytoplasmic 60 kDa isoform, but not isoforms PR-A, PR-B or PR-S. CONCLUSION: The major PR isoform in the amnion, chorion and placenta is a 60 kDa protein that could be PR-C, suggesting that the cytoplasmic isoform has a specific role in extra-embryonic tissues and may be involved in the regulation of human parturition. PMID- 19284644 TI - Which preventive measures might protect health care workers from SARS? AB - BACKGROUND: Despite the use of a series of preventive measures, a high incidence of severe acute respiratory syndrome (SARS) was observed among health care workers (HCWs) during the SARS epidemic. This study aimed to determine which preventive measures may have been effective in protecting HCWs from infection, and which were not effective. METHODS: A retrospective study was performed among 758 'frontline' health care workers who cared for SARS patients at the Second Affiliated Hospital and the Third Affiliated Hospital of Sun Yat-sen University. The HCWs with IgG against SARS and those without IgG against SARS were respectively defined as the "case group" and the "control group", and logistic regression was conducted to explore the risk factors for SARS infection in HCWs. RESULTS: After adjusting for age, gender, marital status, educational level, professional title, and the department in which an individual worked, the results of a multivariate logistic regression analysis indicated that incidence of SARS among HCWs was significantly and positively associated with: performing tracheal intubations for SARS patients, methods used for air ventilation in wards, avoiding face-to-face interaction with SARS patients, the number of pairs of gloves worn by HCWs, and caring for serious SARS cases. CONCLUSION: Some measures, particularly good air ventilation in SARS wards, may be effective in minimizing or preventing SARS transmission among HCWs in hospitals. PMID- 19284645 TI - Clinical audit of core podiatry treatment in the NHS. AB - BACKGROUND: Core podiatry involves treatment of the nails, corns and callus and also giving footwear and foot health advice. Though it is an integral part of current podiatric practice little evidence is available to support its efficacy in terms of research and audit data. This information is important in order to support the current NHS commissioning process where services are expected to provide data on standards including outcomes. This study aimed to increase the evidence base for this area of practice by conducting a multi-centre audit in 8 NHS podiatry departments over a 1-year period. METHODS: The outcome measure used in this audit was the Podiatry Health Questionnaire which is a self completed short measure of foot health including a pain visual analogue scale and a section for the podiatrist to rate an individual's foot health based on their podiatric problems. The patient questionnaire was completed by individuals prior to receiving podiatry care and then 2 weeks after treatment to assess the effect of core podiatry in terms of pain and foot health. RESULTS: 1047 patients completed both questionnaires, with an age range from 26-95 years and a mean age of 72.9 years. The podiatrists clinical rating at baseline showed 75% of patients had either slight or moderate podiatric problems. The differences in questionnaire and visual analogue scores before and after treatment were determined according to three categories - better, same, worse and 75% of patients' scores either remained the same or improved after core podiatry treatment. A student t-test showed a statistical significant difference in pre and post treatment scores where P < 0.001, though the confidence interval indicated that the improvement was relatively small. CONCLUSION: Core podiatry has been shown to sustain or improve foot health and pain in 75% of the patients taking part in the audit. Simple outcome measures including pain scales should be used routinely in podiatric practice to assess the affect of different aspects of treatments and improve the evidence base for podiatry. PMID- 19284646 TI - The complete mitochondrial genome of the house dust mite Dermatophagoides pteronyssinus (Trouessart): a novel gene arrangement among arthropods. AB - BACKGROUND: The apparent scarcity of available sequence data has greatly impeded evolutionary studies in Acari (mites and ticks). This subclass encompasses over 48,000 species and forms the largest group within the Arachnida. Although mitochondrial genomes are widely utilised for phylogenetic and population genetic studies, only 20 mitochondrial genomes of Acari have been determined, of which only one belongs to the diverse order of the Sarcoptiformes. In this study, we describe the mitochondrial genome of the European house dust mite Dermatophagoides pteronyssinus, the most important member of this largely neglected group. RESULTS: The mitochondrial genome of D. pteronyssinus is a circular DNA molecule of 14,203 bp. It contains the complete set of 37 genes (13 protein coding genes, 2 rRNA genes and 22 tRNA genes), usually present in metazoan mitochondrial genomes. The mitochondrial gene order differs considerably from that of other Acari mitochondrial genomes. Compared to the mitochondrial genome of Limulus polyphemus, considered as the ancestral arthropod pattern, only 11 of the 38 gene boundaries are conserved. The majority strand has a 72.6% AT content but a GC-skew of 0.194. This skew is the reverse of that normally observed for typical animal mitochondrial genomes. A microsatellite was detected in a large non-coding region (286 bp), which probably functions as the control region. Almost all tRNA genes lack a T-arm, provoking the formation of canonical cloverleaf tRNA-structures, and both rRNA genes are considerably reduced in size. Finally, the genomic sequence was used to perform a phylogenetic study. Both maximum likelihood and Bayesian inference analysis clustered D. pteronyssinus with Steganacarus magnus, forming a sistergroup of the Trombidiformes. CONCLUSION: Although the mitochondrial genome of D. pteronyssinus shares different features with previously characterised Acari mitochondrial genomes, it is unique in many ways. Gene order is extremely rearranged and represents a new pattern within the Acari. Both tRNAs and rRNAs are truncated, corroborating the theory of the functional co-evolution of these molecules. Furthermore, the strong and reversed GC- and AT-skews suggest the inversion of the control region as an evolutionary event. Finally, phylogenetic analysis using concatenated mt gene sequences succeeded in recovering Acari relationships concordant with traditional views of phylogeny of Acari. PMID- 19284647 TI - Transanal endoscopic microsurgery versus endoscopic mucosal resection for large rectal adenomas (TREND-study). AB - BACKGROUND: Recent non-randomized studies suggest that extended endoscopic mucosal resection (EMR) is equally effective in removing large rectal adenomas as transanal endoscopic microsurgery (TEM). If equally effective, EMR might be a more cost-effective approach as this strategy does not require expensive equipment, general anesthesia and hospital admission. Furthermore, EMR appears to be associated with fewer complications.The aim of this study is to compare the cost-effectiveness and cost-utility of TEM and EMR for the resection of large rectal adenomas. METHODS/DESIGN: Multicenter randomized trial among 15 hospitals in the Netherlands. Patients with a rectal adenoma > or = 3 cm, located between 1 15 cm ab ano, will be randomized to a TEM- or EMR-treatment strategy. For TEM, patients will be treated under general anesthesia, adenomas will be dissected en bloc by a full-thickness excision, and patients will be admitted to the hospital. For EMR, no or conscious sedation is used, lesions will be resected through the submucosal plane in a piecemeal fashion, and patients will be discharged from the hospital. Residual adenoma that is visible during the first surveillance endoscopy at 3 months will be removed endoscopically in both treatment strategies and is considered as part of the primary treatment. Primary outcome measure is the proportion of patients with recurrence after 3 months. Secondary outcome measures are: 2) number of days not spent in hospital from initial treatment until 2 years afterwards; 3) major and minor morbidity; 4) disease specific and general quality of life; 5) anorectal function; 6) health care utilization and costs. A cost-effectiveness and cost-utility analysis of EMR against TEM for large rectal adenomas will be performed from a societal perspective with respectively the costs per recurrence free patient and the cost per quality adjusted life year as outcome measures. Based on comparable recurrence rates for TEM and EMR of 3.3% and considering an upper-limit of 10% for EMR to be non inferior (beta-error 0.2 and one-sided alpha-error 0.05), 89 patients are needed per group. DISCUSSION: The TREND study is the first randomized trial evaluating whether TEM or EMR is more cost-effective for the treatment of large rectal adenomas. TRIAL REGISTRATION NUMBER: (trialregister.nl) NTR1422. PMID- 19284648 TI - FcgammaRIIa (CD32) polymorphism and anti-malarial IgG subclass pattern among Fulani and sympatric ethnic groups living in eastern Sudan. AB - BACKGROUND: A SNP at position 131, in the FcgammaRIIa gene, affects the binding of the different IgG subclasses and may influence the clinical variation seen in patients with falciparum malaria. This study confirms and extends previous findings, analysing the FcgammaRIIa (CD32) polymorphism in relation to the IgG subclass distribution seen among two sympatric tribes living in eastern Sudan, characterized by marked differences in susceptibility to Plasmodium falciparum malaria. METHODS: Two hundred and fifty Fulani subjects living in an area of meso endemic P. falciparum malaria infection were genotyped for the FcgammaRIIa-131 polymorphism. For comparison, 101 non-Fulani donors - (Masaleit, Hausa and Four) living in the same study area, were genotyped. The levels of plasma antibodies (IgG and subclasses) to four malaria antigens (AMA-1, MSP 2 - 3D7 & FC27, Pf332 C231) were measured using indirect enzyme-linked immunosorbent assays. RESULTS: The FcgammaRIIa-H/H131 genotype was found to be significantly more prevalent in the Fulani as compared to the non-Fulani ethnic groups (36.0% for Fulani versus 17.8% for non-Fulani, adjusted OR 3.10, 95% CI 1.61-5.97, P value < 0.001). The Fulani showed lower anti-malarial IgG1 and IgG3 antibody levels as compared to the non-Fulani and higher levels of IgG2 antibodies. CONCLUSION: The FcgammaRIIa H/H131 genotype and H131 allele is at higher frequency in the Fulani ethnic group. The H/H131 genotype was consistently associated with higher levels of anti malarial IgG2 and IgG3 antibodies, while the R/R131 genotype was associated with higher levels of IgG1 antibodies. PMID- 19284649 TI - Differential patterns of reactive oxygen species and antioxidative mechanisms during atrazine injury and sucrose-induced tolerance in Arabidopsis thaliana plantlets. AB - BACKGROUND: Besides being essential for plant structure and metabolism, soluble carbohydrates play important roles in stress responses. Sucrose has been shown to confer to Arabidopsis seedlings a high level of tolerance to the herbicide atrazine, which causes reactive oxygen species (ROS) production and oxidative stress. The effects of atrazine and of exogenous sucrose on ROS patterns and ROS scavenging systems were studied. Simultaneous analysis of ROS contents, expression of ROS-related genes and activities of ROS-scavenging enzymes gave an integrative view of physiological state and detoxifying potential under conditions of sensitivity or tolerance. RESULTS: Toxicity of atrazine could be related to inefficient activation of singlet oxygen (1O2) quenching pathways leading to 1O2 accumulation. Atrazine treatment also increased hydrogen peroxide (H2O2) content, while reducing gene expressions and enzymatic activities related to two major H2O2-detoxification pathways. Conversely, sucrose-protected plantlets in the presence of atrazine exhibited efficient 1O2 quenching, low 1O2 accumulation and active H2O2-detoxifying systems. CONCLUSION: In conclusion, sucrose protection was in part due to activation of specific ROS scavenging systems with consequent reduction of oxidative damages. Importance of ROS combination and potential interferences of sucrose, xenobiotic and ROS signalling pathways are discussed. PMID- 19284650 TI - TLR9 polymorphisms in African populations: no association with severe malaria, but evidence of cis-variants acting on gene expression. AB - BACKGROUND: During malaria infection the Toll-like receptor 9 (TLR9) is activated through induction with plasmodium DNA or another malaria motif not yet identified. Although TLR9 activation by malaria parasites is well reported, the implication to the susceptibility to severe malaria is not clear. The aim of this study was to assess the contribution of genetic variation at TLR9 to severe malaria. METHODS: This study explores the contribution of TLR9 genetic variants to severe malaria using two approaches. First, an association study of four common single nucleotide polymorphisms was performed on both family- and population-based studies from Malawian and Gambian populations (n>6000 individual). Subsequently, it was assessed whether TLR9 expression is affected by cis-acting variants and if these variants could be mapped. For this work, an allele specific expression (ASE) assay on a panel of HapMap cell lines was carried out. RESULTS: No convincing association was found with polymorphisms in TLR9 for malaria severity, in either Gambian or Malawian populations, using both case-control and family based study designs. Using an allele specific expression assay it was observed that TLR9 expression is affected by cis-acting variants, these results were replicated in a second experiment using biological replicates. CONCLUSION: By using the largest cohorts analysed to date, as well as a standardized phenotype definition and study design, no association of TLR9 genetic variants with severe malaria was found. This analysis considered all common variants in the region, but it is remains possible that there are rare variants with association signals. This report also shows that TLR9 expression is potentially modulated through cis-regulatory variants, which may lead to differential inflammatory responses to infection between individuals. PMID- 19284651 TI - Increased expression of heat shock protein 105 in rat uterus of early pregnancy and its significance in embryo implantation. AB - BACKGROUND: Heat shock proteins (Hsps) are a set of highly conserved proteins, Hsp105, has been suggested to play a role in reproduction. METHODS: Spatio temporal expression of Hsp105 in rat uterus during peri-implantation period was examined by immunohistochemistry and Western blot, pseudopregnant uterus was used as control. Injection of antisense oligodeoxynucleotides to Hsp105 into pregnant rat uteri was carried out to look at effect of Hsp105 on embryo implantation. RESULTS: Expression of Hsp105 was mainly in the luminal epithelium on day 1 of pregnancy, and reached a peak level on day 5, whereas in stroma cells, adjacent to the implanting embryo, the strongest expression of Hsp105 was observed on day 6. The immunostaining profile in the uterus was consistent with that obtained by Western blot in the early pregnancy. In contrast, no obvious peak level of Hsp105 was observed in the uterus of pseudopregnant rat on day 5 or day 6. Furthermore, injection of antisense oligodeoxynucleotides to Hsp105 into the rat uterine horn on day 3 of pregnancy obviously suppressed the protein expression as expected and reduced number of the implanted embryos as compared with the control. CONCLUSION: Temporal and spatial changes in Hsp105 expression in pregnant rat uterus may play a physiological role in regulating embryo implantation. PMID- 19284652 TI - A BAC-bacterial recombination method to generate physically linked multiple gene reporter DNA constructs. AB - BACKGROUND: Reporter gene mice are valuable animal models for biological research providing a gene expression readout that can contribute to cellular characterization within the context of a developmental process. With the advancement of bacterial recombination techniques to engineer reporter gene constructs from BAC genomic clones and the generation of optically distinguishable fluorescent protein reporter genes, there is an unprecedented capability to engineer more informative transgenic reporter mouse models relative to what has been traditionally available. RESULTS: We demonstrate here our first effort on the development of a three stage bacterial recombination strategy to physically link multiple genes together with their respective fluorescent protein (FP) reporters in one DNA fragment. This strategy uses bacterial recombination techniques to: (1) subclone genes of interest into BAC linking vectors, (2) insert desired reporter genes into respective genes and (3) link different gene reporters together. As proof of concept, we have generated a single DNA fragment containing the genes Trap, Dmp1, and Ibsp driving the expression of ECFP, mCherry, and Topaz FP reporter genes, respectively. Using this DNA construct, we have successfully generated transgenic reporter mice that retain two to three gene readouts. CONCLUSION: The three stage methodology to link multiple genes with their respective fluorescent protein reporter works with reasonable efficiency. Moreover, gene linkage allows for their common chromosomal integration into a single locus. However, the testing of this multi-reporter DNA construct by transgenesis does suggest that the linkage of two different genes together, despite their large size, can still create a positional effect. We believe that gene choice, genomic DNA fragment size and the presence of endogenous insulator elements are critical variables. PMID- 19284653 TI - Implication of scavenger receptors in the interactions between diesel exhaust particles and immature or mature dendritic cells. AB - BACKGROUND: The exposure to pollutants such as diesel exhaust particles (DEP) is associated with an increased incidence of respiratory diseases. However, the mechanisms by which DEP have an effect on human health are not completely understood. In addition to their action on macrophages and airway epithelial cells, DEP also modulate the functions of dendritic cells (DC). These professional antigen-presenting cells are able to discriminate unmodified self from non-self thanks to pattern recognition receptors such as the Toll like Receptors (TLR) and Scavenger Receptors (SR). SR were originally identified by their ability to bind and internalize modified lipoproteins and microorganisms but also particles and TLR agonists. In this study, we assessed the implication of SR in the effects of DEP associated or not with TLR agonists on monocyte derived DC (MDDC). For this, we studied the regulation of CD36, CXCL16, LOX-1, SR A1 and SR-B1 expression on MDDC treated with DEP associated or not with TLR2, 3 and 4 ligands. Then, the capacity of SR ligands (dextran sulfate and maleylated ovalbumin) to block the effects of DEP on the function of lipopolysaccharide (LPS)-activated DC has been evaluated. RESULTS: Our data demonstrate that TLR2 agonists mainly augmented CXCL16, LOX-1 and SR-B1 expression whereas DEP alone had only a weak effect. Interestingly, DEP modulated the action of TLR2 and TLR4 ligands on the expression of LOX-1 and SR-B1. Pretreatment with the SR ligand maleylated-ovalbumin but not dextran sulfate inhibited the endocytosis of DEP by MDDC. Moreover, this SR ligand blocked the effect by DEP at low dose (1 mug/ml) on MDDC phenotype (a decrease of CD86 and HLA-DR expression) and on the secretion of CXCL10, IL-12 and TNF-alpha. In contrast, the decrease of IL-12 and CXCL10 secretion and the generation of oxygen metabolite induced by DEP at 10 mug/ml was not affected by SR ligands CONCLUSION: Our results show for the first time that the modulation of DC functions by DEP implicates SR. TLR agonists upregulated SR expression in contrast to DEP. Interfering with the expression and/or the function of SR might be one way to limit the impact of DEP on lung immune response. PMID- 19284654 TI - Discovering genes associated with dormancy in the monogonont rotifer Brachionus plicatilis. AB - BACKGROUND: Microscopic monogonont rotifers, including the euryhaline species Brachionus plicatilis, are typically found in water bodies where environmental factors restrict population growth to short periods lasting days or months. The survival of the population is ensured via the production of resting eggs that show a remarkable tolerance to unfavorable conditions and remain viable for decades. The aim of this study was to generate Expressed Sequence Tags (ESTs) for molecular characterisation of processes associated with the formation of resting eggs, their survival during dormancy and hatching. RESULTS: Four normalized and four subtractive libraries were constructed to provide a resource for rotifer transcriptomics associated with resting-egg formation, storage and hatching. A total of 47,926 sequences were assembled into 18,000 putative transcripts and analyzed using both Blast and GO annotation. About 28-55% (depending on the library) of the clones produced significant matches against the Swissprot and Trembl databases. Genes known to be associated with desiccation tolerance during dormancy in other organisms were identified in the EST libraries. These included genes associated with antioxidant activity, low molecular weight heat shock proteins and Late Embryonic Abundant (LEA) proteins. Real-time PCR confirmed that LEA transcripts, small heat-shock proteins and some antioxidant genes were upregulated in resting eggs, therefore suggesting that desiccation tolerance is a characteristic feature of resting eggs even though they do not necessarily fully desiccate during dormancy. The role of trehalose in resting-egg formation and survival remains unclear since there was no significant difference between resting-egg producing females and amictic females in the expression of the tps-1 gene. In view of the absence of vitellogenin transcripts, matches to lipoprotein lipase proteins suggest that, similar to the situation in dipterans, these proteins may serve as the yolk proteins in rotifers. CONCLUSION: The 47,926 ESTs expand significantly the current sequence resource of B. plicatilis. It describes, for the first time, genes putatively associated with resting eggs and will serve as a database for future global expression experiments, particularly for the further identification of dormancy related genes. PMID- 19284655 TI - Inhibition of S6K1 accounts partially for the anti-inflammatory effects of the arginase inhibitor L-norvaline. AB - BACKGROUND: Pharmacological inhibition of endothelial arginase-II has been shown to improve endothelial nitric oxide synthase (eNOS) function and reduce atherogenesis in animal models. We investigated whether the endothelial arginase II is involved in inflammatory responses in endothelial cells. METHODS: Human endothelial cells were isolated from umbilical veins and stimulated with TNFalpha (10 ng/ml) for 4 hours. Endothelial expression of the inflammatory molecules i.e. vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and E-selectin were assessed by immunoblotting. RESULTS: The induction of the expression of endothelial VCAM-1, ICAM-1 and E-selectin by TNFalpha was concentration-dependently reduced by incubation of the endothelial cells with the arginase inhibitor L-norvaline. However, inhibition of arginase by another arginase inhibitor S-(2-boronoethyl)-L-cysteine (BEC) had no effects. To confirm the role of arginase-II (the prominent isoform expressed in HUVECs) in the inflammatory responses, adenoviral mediated siRNA silencing of arginase-II knocked down the arginase II protein level, but did not inhibit the up-regulation of the adhesion molecules. Moreover, the inhibitory effect of L-norvaline was not reversed by the NOS inhibitor L-NAME and L-norvaline did not interfere with TNFalpha-induced activation of NF-kappaB, JNK, p38mapk, while it inhibited p70s6k (S6K1) activity. Silencing S6K1 prevented up-regulation of E-selectin, but not that of VCAM-1 or ICAM-1 induced by TNFalpha. CONCLUSION: The arginase inhibitor L-norvaline exhibits anti-inflammatory effects independently of inhibition of arginase in human endothelial cells. The anti-inflammatory properties of L norvaline are partially attributable to its ability to inhibit S6K1. PMID- 19284657 TI - Early cellular signaling responses to axonal injury. AB - BACKGROUND: We have used optic nerve injury as a model to study early signaling events in neuronal tissue following axonal injury. Optic nerve injury results in the selective death of retinal ganglion cells (RGCs). The time course of cell death takes place over a period of days with the earliest detection of RGC death at about 48 hr post injury. We hypothesized that in the period immediately following axonal injury, there are changes in the soma that signal surrounding glia and neurons and that start programmed cell death. In the current study, we investigated early changes in cellular signaling and gene expression that occur within the first 6 hrs post optic nerve injury. RESULTS: We found evidence of cell to cell signaling within 30 min of axonal injury. We detected differences in phosphoproteins and gene expression within the 6 hrs time period. Activation of TNFalpha and glutamate receptors, two pathways that can initiate cell death, begins in RGCs within 6 hrs following axonal injury. Differential gene expression at 6 hrs post injury included genes involved in cytokine, neurotrophic factor signaling (Socs3) and apoptosis (Bax). CONCLUSION: We interpret our studies to indicate that both neurons and glia in the retina have been signaled within 30 min after optic nerve injury. The signals are probably initiated by the RGC soma. In addition, signals activating cellular death pathways occur within 6 hrs of injury, which likely lead to RGC degeneration. PMID- 19284658 TI - New fluoroscopic imaging technique for investigation of 6DOF knee kinematics during treadmill gait. AB - INTRODUCTION: This report presents a new imaging technique for non-invasive study of six degrees of freedom (DOF) knee kinematics during treadmill gait. MATERIALS AND METHODS: A treadmill was integrated into a dual fluoroscopic imaging system (DFIS) to formulate a gait analysis system. To demonstrate the application of the system, a healthy subject walked on the treadmill at four different speeds (1.5, 2.0, 2.5 and 3.0 MPH) while the DFIS captured the knee motion during three strides under each speed. Characters of knee joint motion were analyzed in 6DOF during the treadmill walking. RESULTS: The speed of the knee motion was lower than that of the treadmill. Flexion amplitudes increased with increasing walking speed. Motion patterns in other DOF were not affected by increase in walking speed. The motion character was repeatable under each treadmill speed. CONCLUSION: The presented technique can be used to accurately measure the 6DOF knee kinematics at normal walking speeds. PMID- 19284656 TI - Mechanisms of the noxious inflammatory cycle in cystic fibrosis. AB - Multiple evidences indicate that inflammation is an event occurring prior to infection in patients with cystic fibrosis. The self-perpetuating inflammatory cycle may play a pathogenic part in this disease. The role of the NF-kappaB pathway in enhanced production of inflammatory mediators is well documented. The pathophysiologic mechanisms through which the intrinsic inflammatory response develops remain unclear. The unfolded mutated protein cystic fibrosis transmembrane conductance regulator (CFTRDeltaF508), accounting for this pathology, is retained in the endoplasmic reticulum (ER), induces a stress, and modifies calcium homeostasis. Furthermore, CFTR is implicated in the transport of glutathione, the major antioxidant element in cells. CFTR mutations can alter redox homeostasis and induce an oxidative stress. The disturbance of the redox balance may evoke NF-kappaB activation and, in addition, promote apoptosis. In this review, we examine the hypotheses of the integrated pathogenic processes leading to the intrinsic inflammatory response in cystic fibrosis. PMID- 19284659 TI - EXACKTE(2): exploiting the clinical consultation as a knowledge transfer and exchange environment: a study protocol. AB - BACKGROUND: While the evidence suggests that the way physicians provide information to patients is crucial in helping patients decide upon a course of action, the field of knowledge translation and exchange (KTE) is silent about how the physician and the patient influence each other during clinical interactions and decision-making. Consequently, based on a novel relationship-centered model, EXACKTE(2) (EXploiting the clinicAl Consultation as a Knowledge Transfer and Exchange Environment), this study proposes to assess how patients and physicians influence each other in consultations. METHODS: We will employ a cross-sectional study design involving 300 pairs of patients and family physicians from two primary care practice-based research networks. The consultation between patient and physician will be audio-taped and transcribed. Following the consultation, patients and physicians will complete a set of questionnaires based on the EXACKTE(2) model. All questionnaires will be similar for patients and physicians. These questionnaires will assess the key concepts of our proposed model based on the essential elements of shared decision-making (SDM): definition and explanation of problem; presentation of options; discussion of pros and cons; clarification of patient values and preferences; discussion of patient ability and self-efficacy; presentation of doctor knowledge and recommendation; and checking and clarifying understanding. Patients will be contacted by phone two weeks later and asked to complete questionnaires on decisional regret and quality of life. The analysis will be conducted to compare the key concepts in the EXACKTE(2) model between patients and physicians. It will also allow the assessment of how patients and physicians influence each other in consultations. DISCUSSION: Our proposed model, EXACKTE(2), is aimed at advancing the science of KTE based on a relationship process when decision-making has to take place. It fosters a new KTE paradigm by putting forward a relationship-centered perspective and has the potential to reveal unknown mechanisms that underline effective KTE in clinical contexts. This will result in better understanding of the mechanisms that may promote a new generation of knowledge transfer strategies. PMID- 19284660 TI - The role of conversation in health care interventions: enabling sensemaking and learning. AB - BACKGROUND: Those attempting to implement changes in health care settings often find that intervention efforts do not progress as expected. Unexpected outcomes are often attributed to variation and/or error in implementation processes. We argue that some unanticipated variation in intervention outcomes arises because unexpected conversations emerge during intervention attempts. The purpose of this paper is to discuss the role of conversation in shaping interventions and to explain why conversation is important in intervention efforts in health care organizations. We draw on literature from sociolinguistics and complex adaptive systems theory to create an interpretive framework and develop our theory. We use insights from a fourteen-year program of research, including both descriptive and intervention studies undertaken to understand and assist primary care practices in making sustainable changes. We enfold these literatures and these insights to articulate a common failure of overlooking the role of conversation in intervention success, and to develop a theoretical argument for the importance of paying attention to the role of conversation in health care interventions. DISCUSSION: Conversation between organizational members plays an important role in the success of interventions aimed at improving health care delivery. Conversation can facilitate intervention success because interventions often rely on new sensemaking and learning, and these are accomplished through conversation. Conversely, conversation can block the success of an intervention by inhibiting sensemaking and learning. Furthermore, the existing relationship contexts of an organization can influence these conversational possibilities. We argue that the likelihood of intervention success will increase if the role of conversation is considered in the intervention process. SUMMARY: The generation of productive conversation should be considered as one of the foundations of intervention efforts. We suggest that intervention facilitators consider the following actions as strategies for reducing the barriers that conversation can present and for using conversation to leverage improvement change: evaluate existing conversation and relationship systems, look for and leverage unexpected conversation, create time and space where conversation can unfold, use conversation to help people manage uncertainty, use conversation to help reorganize relationships, and build social interaction competence. PMID- 19284661 TI - Statistical language learning in neonates revealed by event-related brain potentials. AB - BACKGROUND: Statistical learning is a candidate for one of the basic prerequisites underlying the expeditious acquisition of spoken language. Infants from 8 months of age exhibit this form of learning to segment fluent speech into distinct words. To test the statistical learning skills at birth, we recorded event-related brain responses of sleeping neonates while they were listening to a stream of syllables containing statistical cues to word boundaries. RESULTS: We found evidence that sleeping neonates are able to automatically extract statistical properties of the speech input and thus detect the word boundaries in a continuous stream of syllables containing no morphological cues. Syllable specific event-related brain responses found in two separate studies demonstrated that the neonatal brain treated the syllables differently according to their position within pseudowords. CONCLUSION: These results demonstrate that neonates can efficiently learn transitional probabilities or frequencies of co-occurrence between different syllables, enabling them to detect word boundaries and in this way isolate single words out of fluent natural speech. The ability to adopt statistical structures from speech may play a fundamental role as one of the earliest prerequisites of language acquisition. PMID- 19284662 TI - Potentially avoidable inpatient nights among warfarin receiving patients; an audit of a single university teaching hospital. AB - BACKGROUND: Warfarin is an oral anticoagulant (OAT) that needs active management to ensure therapeutic range. Initial management is often carried out as an inpatient, though not requiring inpatient facilities. This mismatch results in financial costs which could be directed more efficaciously. The extent of this has previously been unknown. Here we aim to calculate the potential number of bed nights which may be saved among those being dose optimized as inpatients and examine associated factors. METHODS: A 6 week prospective audit of inpatients receiving OAT, at Cork University Hospital, was carried out. The study period was from 11th June 2007 to 20th July 2007. Data was collected from patient's medications prescription charts, medical record files, and computerised haematology laboratory records. The indications for OAT, the patient laboratory coagulation results and therapeutic intervals along with patient demographics were analysed. The level of potentially avoidable inpatient nights in those receiving OAT in hospital was calculated and the potential cost savings quantified. Potential avoidable bed nights were defined as patients remaining in hospital for the purpose of optimizing OAT dosage, while receiving subtherapeutic or therapeutic OAT (being titred up to therapeutic levels) and co-administered covering low molecular weight heparin, and requiring no other active care. The average cost of euro638 was taken as the per night hospital stay cost for a non Intensive Care bed. Ethical approval was granted from the Ethical Committee of the Cork Teaching Hospitals, Cork, Ireland. RESULTS: A total of 158 patients were included in the audit. There was 94 men (59.4%) and 64 women (40.6%). The mean age was 67.8 years, with a median age of 70 years.Atrial Fibrillation (43%, n = 70), followed by aortic valve replacement (15%, n = 23) and pulmonary emboli (11%, n = 18) were the commonest reasons for prescribing OAT. 54% had previously been prescribed OAT prior to current admission.It was confirmed that, there are potentially avoidable nights in patients receiving OAT. The majority of this group were those being commenced on OAT for the first time (p = 0.00002), in the specialities of Cardiology, Cardiothoracic surgery and Care of the Elderly. The potential number of bed nights to be saved is 13 per week for the hospital or 1.1 bed nights per 10,000 general hospital admissions. These were predominantly weekday nights. The estimated cost of avoidable inpatient OAT dose optimization was approximately euro8300 per week. CONCLUSION: With rising costs and the increasing demands for acute hospital beds, alterations to inpatient management for this group of patients should be considered. Alternatives include increasing the size of current anticoagulation clinics, introduction of POCT (point of care testing) devices and increased GP management. POCT can be justified based upon the publication by Gardiner et al, who showed that 87% of patients find self testing straightforward, 87% were confident in the result they obtained using the devices and 77% preferred self testing. PMID- 19284663 TI - Methodology of a diabetes prevention translational research project utilizing a community-academic partnership for implementation in an underserved Latino community. AB - BACKGROUND: Latinos comprise the largest racial/ethnic group in the United States and have 2-3 times the prevalence of type 2 diabetes mellitus as Caucasians. METHODS AND DESIGN: The Lawrence Latino Diabetes Prevention Project (LLDPP) is a community-based translational research study which aims to reduce the risk of diabetes among Latinos who have a >/= 30% probability of developing diabetes in the next 7.5 years per a predictive equation. The project was conducted in Lawrence, Massachusetts, a predominantly Caribbean-origin urban Latino community. Individuals were identified primarily from a community health center's patient panel, screened for study eligibility, randomized to either a usual care or a lifestyle intervention condition, and followed for one year. Like the efficacious Diabetes Prevention Program (DPP), the LLDPP intervention targeted weight loss through dietary change and increased physical activity. However, unlike the DPP, the LLDPP intervention was less intensive, tailored to literacy needs and cultural preferences, and delivered in Spanish. The group format of the intervention (13 group sessions over 1 year) was complemented by 3 individual home visits and was implemented by individuals from the community with training and supervision by a clinical research nutritionist and a behavioral psychologist. Study measures included demographics, Stern predictive equation components (age, gender, ethnicity, fasting glucose, systolic blood pressure, HDL cholesterol, body mass index, and family history of diabetes), glycosylated hemoglobin, dietary intake, physical activity, depressive symptoms, social support, quality of life, and medication use. Body weight was measured at baseline, 6-months, and one-year; all other measures were assessed at baseline and one-year. All surveys were orally administered in Spanish. RESULTS: A community-academic partnership enabled the successful recruitment, intervention, and assessment of Latinos at risk of diabetes with a one-year study retention rate of 93%. TRIAL REGISTRATION: NCT00810290. PMID- 19284664 TI - Characterization of the epithelial sodium channel alpha subunit coding and non coding transcripts and their corresponding mRNA expression levels in Dahl R versus S rat kidney cortex on normal and high salt diet. AB - AIMS/HYPOTHESIS: The alpha subunit of the amiloride-sensitive epithelial sodium channel (alpha ENaC) is critical for the expression of functional channels. In humans and rats, non functional alternatively spliced forms of alpha ENaC have been proposed to act as negative regulatory components for ENaC. The purpose of this study was to examine the presence and consequently investigate the mRNA expression levels of alternatively spliced forms of alpha ENaC in kidney cortex of Dahl salt-resistant rats (R) versus Dahl salt-sensitive rats (S) on high salt and normal diets. METHODS: Using quantitative RT-PCR strategy, we examined the mRNA expression levels of previously reported alpha ENaC-a and -b alternatively spliced forms in kidney cortex of Dahl S and R rats on normal and four-week high salt diet and compared their corresponding abundance to wildtype alpha ENaC mRNA levels. We identified 2 novel non-coding C-terminus spliced forms and examined their mRNA expression in Dahl R versus S rat kidney cortex. We also tested the presence of five previously reported lung-specific alpha ENaC spliced forms in Dahl rat kidney cortex (CK479583, CK475461, CK364785, CK475819, and CB690980). RESULTS: Previously reported alpha ENaC-a and -b alternatively spliced forms are present in Dahl rat kidney cortex and are significantly higher in Dahl R versus S rats (P < 0.05). Four-week high salt diet significantly increases alpha ENaC-b (P < 0.05), but not alpha ENaC-a transcript abundance in Dahl R, but not S rats. Two non-coding alpha ENaC spliced forms -c and -d are newly identified in the present study, whose levels are comparable in Dahl R and S rats. Compared to alpha ENaC wt, alpha ENaC-a, -c and -d are low abundance transcripts (4 +/- 2, 110 +/- 20, and 10 +/- 2 fold less respectively), in contrast to alpha ENaC-b abundance that exceeds alpha ENaC-wt by 32 +/- 3 fold. We could not identify any of the five previously reported lung-specific alpha ENaC spliced forms (CK479583, CK475461, CK364785, CK475819, and CB690980) in Dahl rat kidney cortex. CONCLUSION/INTERPRETATION: alpha ENaC alternative splicing might regulate alpha ENaC by the formation of coding RNA species (alpha ENaC-a and -b) and non-coding RNA species (alpha ENaC-c and -d). alpha ENaC-a and -b mRNA levels are significantly higher in Dahl R versus S rats. Additionally, alpha ENaC-b is a salt-sensitive transcript whose levels are significantly higher 4-weeks post high salt diet compared to normal salt diet in Dahl R rats. Among the four alpha ENaC transcripts (-a, -b, -c and -d), alpha ENaC-b is a predominant transcript that exceeds alpha ENaC-wt abundance by ~32 fold. alpha ENaC-a and -b spliced forms, particularly, alpha ENaC-b, might potentially act as dominant negative proteins for ENaC activity, thereby rescuing Dahl R rats from developing salt-sensitive hypertension on high salt diet. On the other hand, non-coding alpha ENaC-c and -d might assist alternative splicing, facilitate RNA processing, or regulate alpha ENaC as well as each other. PMID- 19284665 TI - Associations among hair loss, oral sulfur-containing gases, and gastrointestinal and metabolic linked diseases in Japanese elderly men: pilot study. AB - BACKGROUND: Male pattern baldness (MPB), an observable trait, has been reported to be associated with various diseases, such as prostate cancer and cardiovascular disease. Oral sulfur-containing gases have also been suggested to be useful as markers of systemic health condition. However, there are no known reports regarding the associations among MPB, and oral sulfur-containing gases, and systemic health conditions in males. METHODS: We studied 170 male subjects aged either 60 or 65 years old. The degree of MPB was assessed using the Norwood Hamilton Baldness scale. Oral sulfur-containing gases were measured using a compact-designed device. All subjects completed physical and laboratory blood examinations, a face-to-face medical questionnaire, and an oral examination. RESULTS: There were significant differences between the levels of CH3SCH3 and baldness patterns, independent of age. When we analyzed whether the association was linked to systemic health condition, a strong significant association was observed between the level of CH3SCH3 and severe MPB in subjects with gastrointestinal diseases, hypertension, and hypercholesterolemia. CONCLUSION: These results suggest that MPB is associated with the level of CH3SCH3, a sulfur containing gas that causes oral malodor, in elderly Japanese males. Further, the association was intensified by the existence of gastrointestinal tract and metabolic disorders. PMID- 19284666 TI - Genetic polymorphisms in DNA base excision repair gene XRCC1 and the risk of squamous cell carcinoma of the head and neck. AB - BACKGROUND: The genes of base excision repair (BER) pathway have been extensively studied in the association with various human cancers. We performed a case control study to test the association between two common single nucleotide polymorphisms (SNPs) of XRCC1 gene with human head and neck squamous cell carcinoma (HNSCC). METHODS: The genotype analysis of Arg194Trp and Arg399Gln gene polymorphisms for 92 HNSCC patients and 124 controls of cancer free subjects, in Polish population were performed using the PCR-based restriction fragment length polymorphism (PCR-RFLP) with endonuclease MspI. RESULTS: No altered risk has been found individually for these SNPs, however haplotypes analysis showed high association with head and neck cancer. The highest frequency, according to wild type of Arg194Arg and Arg399Arg genotypes, was identified for Arg194Trp-Arg399Arg haplotype (OR, 2.96; 95% CI, 1.01-8.80). CONCLUSION: Finally, we identified the combined Arg194Trp-Arg399Arg genotype of base excision repair gene XRCC1 that was associated with HNSCC and may have an impact on identification of a high-risk cancer population. PMID- 19284667 TI - Factors influencing quality of life in Moroccan postmenopausal women with osteoporotic vertebral fracture assessed by ECOS 16 questionnaire. AB - OBJECTIVE: The aim of the study was to evaluate factors influencing quality of life (QOL) in Moroccan postmenopausal women with osteoporotic vertebral fracture assessed by the Arabic version of ECOS 16 questionnaire. METHODS: 357 postmenopausal women were included in this study. The participants underwent bone mineral density (BMD) measurements by DXA of the lumbar spine and the total hip as well as X-ray examination of the thoraco-lumbar spine to identify subclinical vertebral fractures. Patients were asked to complete a questionnaire on clinical and sociodemographic parameters, and osteoporosis risk factors. The Arabic version of the ECOS16 (Assessment of health related quality of life in osteoporosis questionnaire) was used to assess quality of life. RESULTS: The mean age was 58 +/- 7.8 years, and the mean BMI was 28.3 +/- 4.8 kg/m2. One hundred and eight women (30.1%) were osteoporotic and 46.7% had vertebral fractures. Most were categorized as Grade1 (75%). Three independent factors were associated with a poor quality of life: low educational level (p = 0,01), vertebral fracture (p = 0,03), and history of peripheral fracture (p = 0,006). Worse QOL was observed in the group with vertebral fracture in all domains except "pain": Physical functioning (p = 0,002); Fear of illness (p = 0,001); and Psychosocial functioning (p = 0,007). The number of fractures was a determinant of a low QOL, as indicated by an increased score in physical functioning (p = 0,01), fear of illness (p = 0,007), and total score (p = 0,01) after adjusting on age and educational level. Patients with higher Genant score had low QOL in these two domains too (p = 0,002; p = 0,001 respectively), and in the total score (p = 0,01) after adjusting on age and educational level. CONCLUSION: Our current data showed that the quality of life assessed by the Arabic version of the ECOS 16 questionnaire is decreased in post menopausal women with prevalent vertebral fractures, with the increasing number and the severity of vertebral fractures. PMID- 19284668 TI - Dietary Guidelines should reflect new understandings about adult protein needs. AB - Dietary Guidelines for Americans provide nutrition advice aimed at promoting healthy dietary choices for life-long health and reducing risk of chronic diseases. With the advancing age of the population, the 2010 Dietary Guidelines confront increasing risks for age-related problems of obesity, osteoporosis, type 2 diabetes, Metabolic Syndrome, heart disease, and sarcopenia. New research demonstrates that the meal distribution and amount of protein are important in maintaining body composition, bone health and glucose homeostasis. This editorial reviews the benefits of dietary protein for adult health, addresses omissions in current nutrition guidelines, and offers concepts for improving the Dietary Guidelines. PMID- 19284669 TI - Differences in service utilization and barriers among Blacks, Hispanics, and Whites with drug use disorders. AB - BACKGROUND: Treatment for drug use disorders (DUD) can be effective, but only a small proportion of people with DUD seek or receive treatment. Research on racial and ethnic treatment differences and disparities remains unclear. Understanding racial and ethnic differences and disparities in drug treatment is necessary in order to develop a more effective referral system and to improve the accessibility of treatment. The purpose of the current study was to explore the role of race and ethnicity in service utilization. METHODS: Using data from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), this study examined racial and ethnic differences in use of 14 types of treatment services for DUD and 27 different treatment barriers among persons who met lifetime criteria for a DUD. Multivariate logistic regression analyses were used to examine service utilization and barriers among the racial and ethnic groups, while adjusting for other sociodemographic and clinical variables. RESULTS AND DISCUSSION: Among Blacks, Hispanics and Whites in the overall NESARC sample, approximately 10.5% met criteria for at least one lifetime drug use disorder. Approximately 16.2% of persons with a lifetime DUD received at least one type of service. Overall, this study indicated that Whites were less likely to report receiving help for drug-related problems than Blacks, Blacks used a greater number of different types of services, and no racial and ethnic differences were observed with respect to perceived barriers to drug treatment. However, by examining types of services separately, a complex picture of racial and ethnic differences emerges. Most notably, Whites were most likely to use professional services, whereas Blacks were most likely to use 12-step and clergy. The service use pattern of Hispanics most resembled that of Whites. CONCLUSION: While structural barriers to accessing treatment were observed, broad-based educational programs and interventions that are appropriately targeted to racial and ethnic groups remains an important area for prevention and treatment. PMID- 19284670 TI - A model to optimize public health care and downstage breast cancer in limited resource populations in southern Brazil. (Porto Alegre Breast Health Intervention Cohort). AB - BACKGROUND: Breast cancer (BC) is a major public health problem, with rising incidence in many regions of the globe. Although mortality has recently dropped in developed countries, death rates are still increasing in some developing countries, as seen in Brazil. Among the reasons for this phenomenon are the lack of structured screening programs, a long waiting period between diagnosis and treatment, and lack of access to health services for a large proportion of the Brazilian population. METHODS AND DESIGN: Since 2004, an intervention study in a cohort of women in Southern Brazil, denominated Porto Alegre Breast Health Intervention Cohort, is being conducted in order to test the effectiveness and cost-effectiveness of a model for BC early detection and treatment. In this study, over 4,000 women from underserved communities aged 40 to 69 years are being screened annually with mammography and clinical breast examination performed by a multidisciplinary team, which also involves nutritional counseling and genetic cancer risk assessment. Risk factors for BC development are also being evaluated. Active search of participants by lay community health workers is one of the major features of our program. The accrual of new participants was concluded in 2006 and the study will last for 10 years. The main goal of the study is to demonstrate significant downstaging of BC in an underserved population through proper screening, attaining a higher rate of early-stage BC diagnoses than usually seen in women diagnosed in the Brazilian Public Health System. Preliminary results show a very high BC incidence in this population (117 cases per 100,000 women per year), despite a low prevalence of classical risk factors. DISCUSSION: This study will allow us to test a model of BC early diagnosis and treatment and evaluate its cost-effectiveness in a developing country where the mortality associated with this disease is very high. Also, it might contribute to the evaluation of risk factors in a population with a different ethnic background from that studied in developed countries. If our model is proven effective, it may be replicated in other parts of the globe where BC is also a major public health problem. PMID- 19284671 TI - Coincident frequencies and relative phases among brain activity and hormonal signals. AB - BACKGROUND: Fourier transform is a basic tool for analyzing biological signals and is computed for a finite sequence of data sample. The electroencephalographic (EEG) signals analyzed with this method provide only information based on the frequency range, for short periods. In some cases, for long periods it can be useful to know whether EEG signals coincide or have a relative phase between them or with other biological signals. Some studies have evidenced that sex hormones and EEG signals show oscillations in their frequencies across a period of 28 days; so it seems of relevance to seek after possible patterns relating EEG signals and endogenous sex hormones, assumed as long time-periodic functions to determine their typical periods, frequencies and relative phases. METHODS: In this work we propose a method that can be used to analyze brain signals and hormonal levels and obtain frequencies and relative phases among them. This method involves the application of a discrete Fourier Transform on previously reported datasets of absolute power of brain signals delta, theta, alpha1, alpha2, beta1 and beta2 and the endogenous estrogen and progesterone levels along 28 days. RESULTS: Applying the proposed method to exemplary datasets and comparing each brain signal with both sex hormones signals, we found a characteristic profile of coincident periods and typical relative phases. For the corresponding coincident periods the progesterone seems to be essentially in phase with theta, alpha1, alpha2 and beta1, while delta and beta2 go oppositely. For the relevant coincident periods, the estrogen goes in phase with delta and theta and goes oppositely with alpha2. CONCLUSION: Findings suggest that the procedure applied here provides a method to analyze typical frequencies, or periods and phases between signals with the same period. It generates specific patterns for brain signals and hormones and relations among them. PMID- 19284672 TI - Greater involvement of people living with HIV in health care. AB - Greater Involvement of People Living with HIV/AIDS represents a mobilising and an organising principle for the involvement of people living with HIV in program and policy responses. People with HIV have been at the forefront of designing and implementing effective HIV treatment, care and prevention activities. However, governments and health systems have yet to act to fully harness the potential and resources of people living with HIV in addressing the epidemic.The lives and experiences of people living with HIV highlight the need for a shift in the existing paradigm of disease management. The high prevalence of HIV amongst health care providers in many countries, exacerbated by stigma towards those with HIV in the health care professions, is seriously undermining the capacity of health systems and signals the need to change the current nature of health care delivery. Moreover, the negative experiences of many people with HIV in relation to their health care as well as in their daily social interactions, coupled with the ever-limited current investment in treatment, care and support, demonstrate that the current system is drastically failing the majority of people with HIV. Current health management systems urgently need to be more effectively maximised, to increase the quality of standards of health care systems and services in resource poor countries. An integrated approach to health care based on a human rights framework, grounded in community realities and delivered in partnership and solidarity with people living with HIV, offers the most viable approach to overcoming the crisis of HIV in the health care system. PMID- 19284673 TI - Utilization of public or private health care providers by febrile children after user fee removal in Uganda. AB - BACKGROUND: Despite investments in providing free government health services in Uganda, many caretakers still seek treatment from the drug shops/private clinics. The study aimed to assess determinants for use of government facilities or drug shops/private clinics for febrile illnesses in children under five. METHODS: Structured questionnaires were administered to caretakers in 1078 randomly selected households in the Iganga - Mayuge Demographic Surveillance site. Those with children who had had fever in the previous two weeks and who had sought care from outside the home were interviewed on presenting symptoms and why they chose the provider they went to. Symptoms children presented with and reasons for seeking care from government facilities were compared with those of drug shops/private clinics. RESULTS: Of those who sought care outside the home, 62.7% (286/456) had first gone to drug shops/private clinics and 33.1% (151/456) first went to government facilities. Predictors of having gone to government facilities with a febrile child were child presenting with vomiting (OR 2.07; 95% CI 1.10 - 3.89) and perceiving that the health providers were qualified (OR 10.32; 95% CI 5.84 - 18.26) or experienced (OR 1.93; 95% CI 1.07 - 3.48). Those who took the febrile child to drug shops/private clinics did so because they were going there to get first aid (OR 0.20; 95% CI 0.08 - 0.52). CONCLUSION: Private providers offer 'first aid' to caretakers with febrile children. Government financial assistance to health care providers should not stop at government facilities. Multi-faceted interventions in the private sector and implementation of community case management of febrile children through community medicine distributors could increase the proportion of children who access quality care promptly. PMID- 19284674 TI - The cell nuclei of skeletal muscle cells are transcriptionally active in hibernating edible dormice. AB - BACKGROUND: Skeletal muscle is able to react in a rapid, dynamic way to metabolic and mechanical stimuli. In particular, exposure to either prolonged starvation or disuse results in muscle atrophy. At variance, in hibernating animals muscle atrophy may be scarce or absent after bouts of hibernation i.e., periods of prolonged (months) inactivity and food deprivation, and muscle function is fully preserved at arousal. In this study, myocytes from the quadriceps muscle of euthermic and hibernating edible dormice were investigated by a combination of morphological, morphometrical and immunocytochemical analyses at the light and electron microscopy level. The focus was on cell nuclei and mitochondria, which are highly sensitive markers of changing metabolic rate. RESULTS: Findings presented herein demonstrate that: 1) the general histology of the muscle, inclusive of muscle fibre shape and size, and the ratio of fast and slow fibre types are not affected by hibernation; 2) the fine structure of cytoplasmic and nuclear constituents is similar in euthermia and hibernation but for lipid droplets, which accumulate during lethargy; 3) during hibernation, mitochondria are larger in size with longer cristae, and 4) myonuclei maintain the same amount and distribution of transcripts and transcription factors as in euthermia. CONCLUSION: In this study we demonstrate that skeletal muscle cells of the hibernating edible dormouse maintain their structural and functional integrity in full, even after months in the nest. A twofold explanation for that is envisaged: 1) the maintenance, during hibernation, of low-rate nuclear and mitochondrial activity counterbalancing myofibre wasting, 2) the intensive muscle stimulation (shivering) during periodic arousals in the nest, which would mimic physical exercise. These two factors would prevent muscle atrophy usually occurring in mammals after prolonged starvation and/or inactivity as a consequence of prevailing catabolism. Understanding the mechanisms responsible for skeletal muscle preservation in hibernators could pave the way to prevention and treatment of muscle wasting associated with pathological conditions or ageing as well as life in extreme environments, such as ocean deeps or spaceflights. PMID- 19284675 TI - Mapping carcass and meat quality QTL on Sus Scrofa chromosome 2 in commercial finishing pigs. AB - Quantitative trait loci (QTL) affecting carcass and meat quality located on SSC2 were identified using variance component methods. A large number of traits involved in meat and carcass quality was detected in a commercial crossbred population: 1855 pigs sired by 17 boars from a synthetic line, which where homozygous (A/A) for IGF2. Using combined linkage and linkage disequilibrium mapping (LDLA), several QTL significantly affecting loin muscle mass, ham weight and ham muscles (outer ham and knuckle ham) and meat quality traits, such as Minolta-L* and -b*, ultimate pH and Japanese colour score were detected. These results agreed well with previous QTL-studies involving SSC2. Since our study is carried out on crossbreds, different QTL may be segregating in the parental lines. To address this question, we compared models with a single QTL-variance component with models allowing for separate sire and dam QTL-variance components. The same QTL were identified using a single QTL variance component model compared to a model allowing for separate variances with minor differences with respect to QTL location. However, the variance component method made it possible to detect QTL segregating in the paternal line (e.g. HAMB), the maternal lines (e.g. Ham) or in both (e.g. pHu). Combining association and linkage information among haplotypes improved slightly the significance of the QTL compared to an analysis using linkage information only. PMID- 19284676 TI - The feasibility of gene therapy in the treatment of head and neck cancer. AB - Standard approach to the treatment of head and neck cancer include surgery, chemotherapy, and radiation. More recently, dramatic increases in our knowledge of the molecular and genetic basis of cancer combined with advances in technology have resulted in novel molecular therapies for this disease. In particular, gene therapy, which involves the transfer of genetic material to cells to produce a therapeutic effect, has become a promising approach. Clinical trials concerning gene therapy strategies in head and neck cancer as well as combination of these strategies with chemotherapy and radiation therapy will be discussed. PMID- 19284677 TI - Effects of the number of markers per haplotype and clustering of haplotypes on the accuracy of QTL mapping and prediction of genomic breeding values. AB - The aim of this paper was to compare the effect of haplotype definition on the precision of QTL-mapping and on the accuracy of predicted genomic breeding values. In a multiple QTL model using identity-by-descent (IBD) probabilities between haplotypes, various haplotype definitions were tested i.e. including 2, 6, 12 or 20 marker alleles and clustering base haplotypes related with an IBD probability of > 0.55, 0.75 or 0.95. Simulated data contained 1100 animals with known genotypes and phenotypes and 1000 animals with known genotypes and unknown phenotypes. Genomes comprising 3 Morgan were simulated and contained 74 polymorphic QTL and 383 polymorphic SNP markers with an average r2 value of 0.14 between adjacent markers. The total number of haplotypes decreased up to 50% when the window size was increased from two to 20 markers and decreased by at least 50% when haplotypes related with an IBD probability of > 0.55 instead of > 0.95 were clustered. An intermediate window size led to more precise QTL mapping. Window size and clustering had a limited effect on the accuracy of predicted total breeding values, ranging from 0.79 to 0.81. Our conclusion is that different optimal window sizes should be used in QTL-mapping versus genome-wide breeding value prediction. PMID- 19284678 TI - Detecting parent of origin and dominant QTL in a two-generation commercial poultry pedigree using variance component methodology. AB - INTRODUCTION: Variance component QTL methodology was used to analyse three candidate regions on chicken chromosomes 1, 4 and 5 for dominant and parent-of origin QTL effects. Data were available for bodyweight and conformation score measured at 40 days from a two-generation commercial broiler dam line. One hundred dams were nested in 46 sires with phenotypes and genotypes on 2708 offspring. Linear models were constructed to simultaneously estimate fixed, polygenic and QTL effects. Different genetic models were compared using likelihood ratio test statistics derived from the comparison of full with reduced or null models. Empirical thresholds were derived by permutation analysis. RESULTS: Dominant QTL were found for bodyweight on chicken chromosome 4 and for bodyweight and conformation score on chicken chromosome 5. Suggestive evidence for a maternally expressed QTL for bodyweight and conformation score was found on chromosome 1 in a region corresponding to orthologous imprinted regions in the human and mouse. CONCLUSION: Initial results suggest that variance component analysis can be applied within commercial populations for the direct detection of segregating dominant and parent of origin effects. PMID- 19284679 TI - Head & Neck Oncology: purpose, scope and goals-charting the future. AB - For many years now there has been a growing frustration with the statistics of head and neck cancer. Despite the many advances in diagnosis and therapy, there has been little change in the prognosis for most cancers of the head and neck in the last 50 years, so what is the point of yet another journal? Well, it is not all bad news. PMID- 19284680 TI - Histological changes in intra-oral skin flaps. AB - This review outlines the histological features of intra-oral skin flaps and their pathology as encountered in a routine head and neck diagnostic histopathology service. Problems in recognising and interpreting unfamiliar and complex appearances, and areas of diagnostic and prognostic uncertainty, are highlighted. PMID- 19284681 TI - A fast algorithm for BayesB type of prediction of genome-wide estimates of genetic value. AB - Genomic selection uses genome-wide dense SNP marker genotyping for the prediction of genetic values, and consists of two steps: (1) estimation of SNP effects, and (2) prediction of genetic value based on SNP genotypes and estimates of their effects. For the former step, BayesB type of estimators have been proposed, which assume a priori that many markers have no effects, and some have an effect coming from a gamma or exponential distribution, i.e. a fat-tailed distribution. Whilst such estimators have been developed using Monte Carlo Markov chain (MCMC), here we derive a much faster non-MCMC based estimator by analytically performing the required integrations. The accuracy of the genome-wide breeding value estimates was 0.011 (s.e. 0.005) lower than that of the MCMC based BayesB predictor, which may be because the integrations were performed one-by-one instead of for all SNPs simultaneously. The bias of the new method was opposite to that of the MCMC based BayesB, in that the new method underestimates the breeding values of the best selection candidates, whereas MCMC-BayesB overestimated their breeding values. The new method was computationally several orders of magnitude faster than MCMC based BayesB, which will mainly be advantageous in computer simulations of entire breeding schemes, in cross-validation testing, and practical schemes with frequent re-estimation of breeding values. PMID- 19284682 TI - Review of knee arthroscopy performed under local anesthesia. AB - Local anesthesia for knee arthroscopy is a well documented procedure with diagnostic and therapeutic role. Numerous therapeutic procedures including partial menisectomy, meniscus repair, abrasion chondroplasy, synovectomy, loose body removal can be performed safely and comfortably. Appropriate case selection, anesthetic strategy and technical expertise are the key to smooth and successful surgery. PMID- 19284683 TI - Changes in muscle cell cation regulation and meat quality traits are associated with genetic selection for high body weight and meat yield in broiler chickens. AB - Between-breed genetic variation for muscle and meat quality traits was determined at eight weeks of age in 34 lines of purebred commercial broiler and layer lines and traditional breeds (categories) of chickens. Between-breed genetic variation for plasma ion concentrations and element concentration in muscle dry matter and ash were determined. Plasma from broilers had higher concentrations of Na+, K+, Mg++, total and free Ca++ and lower free:total Ca++ than plasma from layer and traditional lines. Muscle from broilers contained more Na and higher concentrations of K, Mg and Ca per mg of ash but not of dry matter compared with layer and traditional lines. In comparison with layer and traditional lines, broiler genotypes were over three times heavier, their plasma creatine kinase activity (CK), a marker of muscle tissue damage, was higher, their breast muscle colour was lighter (L*) and less red (a*) and yellow (b*) in appearance, the initial and final pH of their muscles were lower, the pH change was higher and their breast muscle was more tender. Thus, genetic selection for broiler traits has markedly altered cation regulation in muscle cells and may be associated with changes in muscle cell function and the development of pathology and meat quality problems. PMID- 19284684 TI - GSE is now an open access journal published by BioMed Central. PMID- 19284685 TI - Ovine progressive pneumonia provirus levels are unaffected by the prion 171R allele in an Idaho sheep flock. AB - Selective breeding of sheep for arginine (R) at prion gene (PRNP) codon 171 confers resistance to classical scrapie. However, other effects of 171R selection are uncertain. Ovine progressive pneumonia/Maedi-Visna virus (OPPV) may infect up to 66% of a flock thus any affect of 171R selection on OPPV susceptibility or disease progression could have major impact on the sheep industry. Hypotheses that the PRNP 171R allele is 1) associated with the presence of OPPV provirus and 2) associated with higher provirus levels were tested in an Idaho ewe flock. OPPV provirus was found in 226 of 358 ewes by quantitative PCR. The frequency of ewes with detectable provirus did not differ significantly among the 171QQ, 171QR, and 171RR genotypes (p > 0.05). Also, OPPV provirus levels in infected ewes were not significantly different among codon 171 genotypes (p > 0.05). These results show that, in the flock examined, the presence of OPPV provirus and provirus levels are not related to the PRNP 171R allele. Therefore, a genetic approach to scrapie control is not expected to increase or decrease the number of OPPV infected sheep or the progression of disease. This study provides further support to the adoption of PRNP 171R selection as a scrapie control measure. PMID- 19284686 TI - Effects of pedigree errors on the efficiency of conservation decisions. AB - Conservation schemes often aim at increasing genetic diversity by minimizing kinship, and the best method to achieve this goal, when pedigree data is available, is to apply optimal contributions. Optimal contributions calculate contributions per animal so that the weighted average mean kinship among candidate parents is minimized. This approach assumes that pedigree data is correct and complete. However, in practice, pedigrees often contain errors: parents are recorded incorrectly or even missing. We used simulations to investigate the effect of these two types of errors on minimizing kinship. Our findings show that a low percentage of wrong parent information reduces the effect of optimal contributions. When the percentage of wrong parent information is above 15%, the population structure and type of errors, should be taken into account before applying optimal contributions. Optimal contributions based on pedigrees with missing parent information hampers conservation of genetic diversity; however, missing parent information can be corrected. It is crucial to know which animals are founders. We strongly recommend that pedigree registration include whether missing parents are either true founders or non-founders. PMID- 19284687 TI - A SMARTT future for sports medicine. PMID- 19284688 TI - Purging of inbreeding depression within the Irish Holstein-Friesian population. AB - The objective of this study was to investigate whether inbreeding depression in milk production or fertility performance has been partially purged due to selection within the Irish Holstein-Friesian population. Classical, ancestral (i.e., the inbreeding of an individual's ancestors according to two different formulae) and new inbreeding coefficients (i.e., part of the classical inbreeding coefficient that is not accounted for by ancestral inbreeding) were computed for all animals. The effect of each coefficient on 305-day milk, fat and protein yield as well as calving interval, age at first calving and survival to second lactation was investigated. Ancestral inbreeding accounting for all common ancestors in the pedigree had a positive effect on 305-day milk and protein yield, increasing yields by 4.85 kg and 0.12 kg, respectively. However, ancestral inbreeding accounting only for those common ancestors, which contribute to the classical inbreeding coefficient had a negative effect on all milk production traits decreasing 305-day milk, fat and protein yields by -8.85 kg, -0.53 kg and 0.33 kg, respectively. Classical, ancestral and new inbreeding generally had a detrimental effect on fertility and survival traits. From this study, it appears that Irish Holstein-Friesians have purged some of their genetic load for milk production through many years of selection based on production alone, while fertility, which has been less intensely selected for in the population demonstrates no evidence of purging. PMID- 19284689 TI - Genetic diversity of a large set of horse breeds raised in France assessed by microsatellite polymorphism. AB - The genetic diversity and structure of horses raised in France were investigated using 11 microsatellite markers and 1679 animals belonging to 34 breeds. Between breed differences explained about ten per cent of the total genetic diversity (Fst = 0.099). Values of expected heterozygosity ranged from 0.43 to 0.79 depending on the breed. According to genetic relationships, multivariate and structure analyses, breeds could be classified into four genetic differentiated groups: warm-blooded, draught, Nordic and pony breeds. Using complementary maximisation of diversity and aggregate diversity approaches, we conclude that particular efforts should be made to conserve five local breeds, namely the Boulonnais, Landais, Merens, Poitevin and Pottok breeds. PMID- 19284690 TI - A mating method accounting for inbreeding and multi-trait selection in dairy cattle populations. AB - Selection in dairy cattle populations usually takes into account both the breed profiles for many traits and their overall estimated breeding values (EBV). This can result in effective contributions of breeding animals departing substantially from contributions optimised for saving future genetic variability. In this work, we propose a mating method that considers not only inbreeding but also the detailed EBV of progeny or the EBV of sires in reference to acceptance thresholds. Penalties were defined for inbreeding and for inadequate EBV profiles. Relative reductions of penalties yielded by any mating design were expressed on a scale ranging from 0 to 1. A value of 0 represented the average performance of random matings and a value of 1 represented the maximal reduction allowed by a specialized, single-penalty, mating design. The core of the method was an adaptative simulated annealing, where the maximized function was the average of both ratios, under the constraints that both relative penalty reductions should be equal and that the within-herd concentration criterion should be equal to a predefined reasonable value. The method was tested on two French dairy cattle populations originating from the same AI organization. The optimised mating design allowed substantial reductions of penalty: 70% and 64% for the Holstein and the Normandy populations, respectively. Thus, this mating method decreased inbreeding and met various demands from breeders. PMID- 19284691 TI - Refeeding syndrome--awareness, prevention and management. AB - BACKGROUND: Refeeding syndrome is an important, yet commonly overlooked condition affecting patients. It occurs when feeding is commenced after a period of starvation. Head and neck cancer patients are at particular risk owing to prolonged periods of poor nutritional intake. This may be from general effects such as cancer anorexia or from more specific problems of dysphagia associated with this group of patients. Awareness of the condition is crucial in identifying patients at risk and taking measures to prevent its occurrence. OBJECTIVES: The aims of this review are to: 1) Highlight the condition and stress the importance of its consideration when admitting head and neck cancer patients. 2) Discuss the pathophysiology behind refeeding syndrome. 3) Review the literature for the best available evidence and guidelines. 4) Highlight the need for further high quality research. CONCLUSION: Refeeding syndrome is potentially fatal, yet is preventable. Awareness and identification of at-risk patients is crucial to improving management. Refeeding syndrome is caused by rapid refeeding after a period of under-nutrition, characterised by hypophosphataemia, electrolyte shifts and has metabolic and clinical complications. High risk patients include the chronically under-nourished and those with little intake for greater than 10 days. Patients with dysphagia are at particular risk. Refeeding should commence at 10 kcal/kg per day in patients at risk, and increased slowly. Thiamine, vitamin B complex and multi-vitamin supplements should be started with refeeding. New NICE guidelines state that pre-feeding correction of electrolyte and fluid deficits is unnecessary, but should be done concurrently with re-feeding. More research in this field is needed as the evidence base is lacking. PMID- 19284692 TI - Genetic architecture of trout from Albania as revealed by mtDNA control region variation. AB - To determine the genetic architecture of trout in Albania, 87 individuals were collected from 19 riverine and lacustrine sites in Albania, FYROM and Greece. All individuals were analyzed for sequence variation in the mtDNA control region. Among fourteen haplotypes detected, four previously unpublished haplotypes, bearing a close relationship to haplotypes of the Adriatic and marmoratus lineages of Salmo trutta, were revealed. Ten previously described haplotypes, characteristic of S. ohridanus, S. letnica and the Adriatic and Mediterranean lineages of S. trutta, were also detected. Haplotypes detected in this study were placed in a well supported branch of S. ohridanus, and a cluster of Mediterranean Adriatic-marmoratus haplotypes, which were further delimited into three subdivisions of Mediterranean, marmoratus, and a previously non-described formation of four Adriatic haplotypes (Balkan cluster). Haplotypes of the Balkan cluster and the other Adriatic haplotypes, do not represent a contiguous haplotype lineage and appear not to be closely related, indicating independent arrivals into the Adriatic drainage and suggesting successive colonization events. Despite the presence of marmoratus haplotypes in Albania, no marbled phenotype was found, confirming previously reported findings that there is no association between this phenotype and marmoratus haplotypes. PMID- 19284693 TI - Genome-assisted prediction of a quantitative trait measured in parents and progeny: application to food conversion rate in chickens. AB - Accuracy of prediction of yet-to-be observed phenotypes for food conversion rate (FCR) in broilers was studied in a genome-assisted selection context. Data consisted of FCR measured on the progeny of 394 sires with SNP information. A Bayesian regression model (Bayes A) and a semi-parametric approach (Reproducing kernel Hilbert Spaces regression, RKHS) using all available SNPs (p = 3481) were compared with a standard linear model in which future performance was predicted using pedigree indexes in the absence of genomic data. The RKHS regression was also tested on several sets of pre-selected SNPs (p = 400) using alternative measures of the information gain provided by the SNPs. All analyses were performed using 333 genotyped sires as training set, and predictions were made on 61 birds as testing set, which were sons of sires in the training set. Accuracy of prediction was measured as the Spearman correlation (_r(S)) between observed and predicted phenotype, with its confidence interval assessed through a bootstrap approach. A large improvement of genome-assisted prediction (up to an almost 4-fold increase in accuracy) was found relative to pedigree index. Bayes A and RKHS regression were equally accurate (_r(S)) = 0.27) when all 3481 SNPs were included in the model. However, RKHS with 400 pre-selected informative SNPs was more accurate than Bayes A with all SNPs. PMID- 19284694 TI - Diagnostic aids in the screening of oral cancer. AB - The World Health Organization has clearly identified prevention and early detection as major objectives in the control of the oral cancer burden worldwide. At the present time, screening of oral cancer and its pre-invasive intra epithelial stages, as well as its early detection, is still largely based on visual examination of the mouth. There is strong available evidence to suggest that visual inspection of the oral mucosa is effective in reducing mortality from oral cancer in individuals exposed to risk factors. Simple visual examination, however, is well known to be limited by subjective interpretation and by the potential, albeit rare, occurrence of dysplasia and early OSCC within areas of normal-looking oral mucosa. As a consequence, adjunctive techniques have been suggested to increase our ability to differentiate between benign abnormalities and dysplastic/malignant changes as well as to identify areas of dysplasia/early OSCC that are not visible to naked eye. These include the use of toluidine blue, brush biopsy, chemiluminescence and tissue autofluorescence. The present paper reviews the evidence supporting the efficacy of the aforementioned techniques in improving the identification of dysplastic/malignant changes of the oral mucosa. We conclude that available studies have shown promising results, but strong evidence to support the use of oral cancer diagnostic aids is still lacking. Further research with clear objectives, well-defined population cohorts, and sound methodology is strongly required. PMID- 19284695 TI - A genetic epidemiological model to describe resistance to an endemic bacterial disease in livestock: application to footrot in sheep. AB - Selection for resistance to an infectious disease not only improves resistance of animals, but also has the potential to reduce the pathogen challenge to contemporaries, especially when the population under selection is the main reservoir of pathogens. A model was developed to describe the epidemiological cycle that animals in affected populations typically go through; viz. susceptible, latently infected, diseased and infectious, recovered and reverting back to susceptible through loss of immunity, and the rates at which animals move from one state to the next, along with effects on the pathogen population. The equilibrium prevalence was estimated as a function of these rates. The likely response to selection for increased resistance was predicted using a quantitative genetic threshold model and also by using epidemiological models with and without reduced pathogen burden. Models were standardised to achieve the same genetic response to one round of selection. The model was then applied to footrot in sheep. The only epidemiological parameters with major impacts for prediction of genetic progress were the rate at which animals recover from infection and the notional reproductive rate of the pathogen. There are few published estimates for these parameters, but plausible values for the rate of recovery would result in a response to selection, in terms of changes in the observed prevalence, double that predicted by purely genetic models in the medium term (e.g. 2-5 generations). PMID- 19284696 TI - Genomic breeding value estimation using nonparametric additive regression models. AB - Genomic selection refers to the use of genomewide dense markers for breeding value estimation and subsequently for selection. The main challenge of genomic breeding value estimation is the estimation of many effects from a limited number of observations. Bayesian methods have been proposed to successfully cope with these challenges. As an alternative class of models, non- and semiparametric models were recently introduced. The present study investigated the ability of nonparametric additive regression models to predict genomic breeding values. The genotypes were modelled for each marker or pair of flanking markers (i.e. the predictors) separately. The nonparametric functions for the predictors were estimated simultaneously using additive model theory, applying a binomial kernel. The optimal degree of smoothing was determined by bootstrapping. A mutation-drift balance simulation was carried out. The breeding values of the last generation (genotyped) was predicted using data from the next last generation (genotyped and phenotyped). The results show moderate to high accuracies of the predicted breeding values. A determination of predictor specific degree of smoothing increased the accuracy. PMID- 19284697 TI - The results of arthroscopic anterior stabilisation of the shoulder using the bioknotless anchor system. AB - BACKGROUND: Shoulder instability is a common condition, particularly affecting a young, active population. Open capsulolabral repair is effective in the majority of cases, however arthroscopic techniques, particularly using suture anchors, are being used with increasing success. METHODS: 15 patients with shoulder instability were operated on by a single surgeon (VK) using BioKnotless anchors (DePuy Mitek, Raynham, MA). The average length of follow-up was 21 months (17 to 31) with none lost to follow-up. Constant scores in both arms, patient satisfaction, activity levels and recurrence of instability was recorded. RESULTS: 80% of patients were satisfied with their surgery. 1 patient suffered a further dislocation and another had recurrent symptomatic instability. The average constant score returned to 84% of that measured in the opposite (unaffected) shoulder. There were no specific post-operative complications encountered. CONCLUSION: In terms of recurrence of symptoms, our results show success rates comparable to other methods of shoulder stabilisation. This technique is safe and surgeons familiar with shoulder arthroscopy will not encounter a steep learning curve. Shoulder function at approximately 2 years post repair was good or excellent in the majority of patients and it was observed that patient satisfaction was correlated more with return to usual activities than recurrence of symptoms. PMID- 19284698 TI - Estimation of prediction error variances via Monte Carlo sampling methods using different formulations of the prediction error variance. AB - Calculation of the exact prediction error variance covariance matrix is often computationally too demanding, which limits its application in REML algorithms, the calculation of accuracies of estimated breeding values and the control of variance of response to selection. Alternatively Monte Carlo sampling can be used to calculate approximations of the prediction error variance, which converge to the true values if enough samples are used. However, in practical situations the number of samples, which are computationally feasible, is limited. The objective of this study was to compare the convergence rate of different formulations of the prediction error variance calculated using Monte Carlo sampling. Four of these formulations were published, four were corresponding alternative versions, and two were derived as part of this study. The different formulations had different convergence rates and these were shown to depend on the number of samples and on the level of prediction error variance. Four formulations were competitive and these made use of information on either the variance of the estimated breeding value and on the variance of the true breeding value minus the estimated breeding value or on the covariance between the true and estimated breeding values. PMID- 19284699 TI - Epiblast-specific Snai1 deletion results in embryonic lethality due to multiple vascular defects. AB - BACKGROUND: Members of the Snail gene family, which encode zinc finger proteins that function as transcriptional repressors, play essential roles during embryonic development in vertebrates. Mouse embryos with conditional deletion of the Snail1 (Snai1) gene in the epiblast, but not in most extraembryonic membranes, exhibit defects in left-right asymmetry specification and migration of mesoderm cells through the posterior primitive streak. Here we describe phenotypic defects that result in death of the mutant embryos by 9.5 days of gestation. FINDINGS: Endothelial cells differentiated in epiblast-specific Snai1 deficient embryos, but formation of an interconnected vascular network was abnormal. To determine whether the observed vascular defects were dependent on disruption of blood flow, we analyzed vascular remodeling in cultured allantois explants from the mutant embryos. Similar vascular defects were observed in the mutant allantois explants. CONCLUSION: These studies demonstrate that lethality in the Snai1-conditional mutant embryos is caused by multiple defects in the cardiovascular system. PMID- 19284700 TI - Rapid intracerebroventricular delivery of Cu-DOTA-etanercept after peripheral administration demonstrated by PET imaging. AB - BACKGROUND: The cytokines interleukin-1 and tumor necrosis factor (TNF), and the cytokine blocker interleukin-1 receptor antagonist, all have been demonstrated to enter the cerebrospinal fluid (CSF) following peripheral administration. Recent reports of rapid clinical improvement in patients with Alzheimer's disease and related forms of dementia following perispinal administration of etanercept, a TNF antagonist, suggest that etanercept also has the ability to reach the brain CSF. To investigate, etanercept was labeled with a positron emitter to enable visualization of its intracranial distribution following peripheral administration by PET in an animal model. FINDINGS: Radiolabeling of etanercept with the PET emitter 64Cu was performed by DOTA (1,4,7,10-tetraazadodecane N,N',N",N"'-tetraacetic acid) conjugation of etanercept, followed by column purification and 64Cu labeling. MicroPET imaging revealed accumulation of 64Cu DOTA-etanercept within the lateral and third cerebral ventricles within minutes of peripheral perispinal administration in a normal rat anesthesized with isoflurane anesthesia, with concentration within the choroid plexus and into the CSF. CONCLUSION: Synthesis of 64Cu-DOTA-etanercept enabled visualization of its intracranial distribution by microPET imaging. MicroPET imaging documented rapid accumulation of 64Cu-DOTA-etanercept within the choroid plexus and the cerebrospinal fluid within the cerebral ventricles of a living rat after peripheral administration. Further study of the effects of etanercept and TNF at the level of the choroid plexus may yield valuable insights into the pathogenesis of Alzheimer's disease. PMID- 19284701 TI - Model for fitting longitudinal traits subject to threshold response applied to genetic evaluation for heat tolerance. AB - A semi-parametric non-linear longitudinal hierarchical model is presented. The model assumes that individual variation exists both in the degree of the linear change of performance (slope) beyond a particular threshold of the independent variable scale and in the magnitude of the threshold itself; these individual variations are attributed to genetic and environmental components. During implementation via a Bayesian MCMC approach, threshold levels were sampled using a Metropolis step because their fully conditional posterior distributions do not have a closed form. The model was tested by simulation following designs similar to previous studies on genetics of heat stress. Posterior means of parameters of interest, under all simulation scenarios, were close to their true values with the latter always being included in the uncertain regions, indicating an absence of bias. The proposed models provide flexible tools for studying genotype by environmental interaction as well as for fitting other longitudinal traits subject to abrupt changes in the performance at particular points on the independent variable scale. PMID- 19284702 TI - A new measure of population structure using multiple single nucleotide polymorphisms and its relationship with FST. AB - BACKGROUND: Large-scale genome-wide association studies are promising for unraveling the genetic basis of complex diseases. Population structure is a potential problem, the effects of which on genetic association studies are controversial. The first step to systematically quantify the effects of population structure is to choose an appropriate measure of population structure for human data. The commonly used measure is Wright's FST. For a set of subpopulations it is generally assumed to be one value of FST. However, the estimates could be different for distinct loci. Since population structure is a concept at the population level, a measure of population structure that utilized the information across loci would be desirable. FINDINGS: In this study we propose an adjusted C parameter according to the sample size from each sub population. The new measure C is based on the c parameter proposed for SNP data, which was assumed to be subpopulation-specific and common for all loci. In this study, we performed extensive simulations of samples with varying levels of population structure to investigate the properties and relationships of both measures. It is found that the two measures generally agree well. CONCLUSION: The new measure simultaneously uses the marker information across the genome. It has the advantage of easy interpretation as one measure of population structure and yet can also assess population differentiation. PMID- 19284703 TI - Genomic selection of purebreds for crossbred performance. AB - BACKGROUND: One of the main limitations of many livestock breeding programs is that selection is in pure breeds housed in high-health environments but the aim is to improve crossbred performance under field conditions. Genomic selection (GS) using high-density genotyping could be used to address this. However in crossbred populations, 1) effects of SNPs may be breed specific, and 2) linkage disequilibrium may not be restricted to markers that are tightly linked to the QTL. In this study we apply GS to select for commercial crossbred performance and compare a model with breed-specific effects of SNP alleles (BSAM) to a model where SNP effects are assumed the same across breeds (ASGM). The impact of breed relatedness (generations since separation), size of the population used for training, and marker density were evaluated. Trait phenotype was controlled by 30 QTL and had a heritability of 0.30 for crossbred individuals. A Bayesian method (Bayes-B) was used to estimate the SNP effects in the crossbred training population and the accuracy of resulting GS breeding values for commercial crossbred performance was validated in the purebred population. RESULTS: Results demonstrate that crossbred data can be used to evaluate purebreds for commercial crossbred performance. Accuracies based on crossbred data were generally not much lower than accuracies based on pure breed data and almost identical when the breeds crossed were closely related breeds. The accuracy of both models (ASGM and BSAM) increased with marker density and size of the training data. Accuracies of both models also tended to decrease with increasing distance between breeds. However the effect of marker density, training data size and distance between breeds differed between the two models. BSAM only performed better than AGSM when the number of markers was small (500), the number of records used for training was large (4000), and when breeds were distantly related or unrelated. CONCLUSION: In conclusion, GS can be conducted in crossbred population and models that fit breed-specific effects of SNP alleles may not be necessary, especially with high marker density. This opens great opportunities for genetic improvement of purebreds for performance of their crossbred descendents in the field, without the need to track pedigrees through the system. PMID- 19284704 TI - Acute aortic dissection: be aware of misdiagnosis. AB - BACKGROUND: Acute aortic dissection (AAD) is a life-threatening condition requiring immediate assessment and therapy. A patient suffering from AAD often presents with an insignificant or irrelevant medical history, giving rise to possible misdiagnosis. The aim of this retrospective study is to address the problem of misdiagnosing AD and the different imaging studies used. METHODS: From January 2000 to December 2004, 49 patients (41 men and 8 women, aged from 18-75 years old) presented to the Emergency Department of our hospital for different reasons and finally diagnosed with AAD. Fifteen of those patients suffered from arterial hypertension, one from giant cell arteritis and another patient from Marfan's syndrome. The diagnosis of AAD was made by chest X-ray, contrast enhanced computed tomography (CT), transthoracic echocardiography (TTE) and coronary angiography. RESULTS: Initial misdiagnosis occurred in fifteen patients (31%) later found to be suffering from AAD. The misdiagnosis was myocardial infarction in 12 patients and cerebral infarction in another three patients. CONCLUSION: Aortic dissection may present with a variety of clinical manifestations, like syncope, chest pain, anuria, pulse deficits, abdominal pain, back pain, or acute congestive heart failure. Nearly a third of the patients found to be suffering from AD, were initially otherwise diagnosed. Key in the management of acute aortic dissection is to maintain a high level of suspicion for this diagnosis. PMID- 19284705 TI - Afferent activity to necklace glomeruli is dependent on external stimuli. AB - BACKGROUND: The main olfactory epithelium (MOE) is a complex organ containing several functionally distinct subpopulations of sensory neurons. One such subpopulation is distinguished by its expression of the guanylyl cyclase GC-D. The axons of GC-D-expressing (GC-D+) neurons innervate 9-15 "necklace" glomeruli encircling the caudal main olfactory bulb (MOB). Chemosensory stimuli for GC-D+ neurons include two natriuretic peptides, uroguanylin and guanylin, and CO2. However, the biologically-relevant source of these chemostimuli is unclear: uroguanylin is both excreted in urine, a rich source of olfactory stimuli for rodents, and expressed in human nasal epithelium; CO2 is present in both inspired and expired air. FINDINGS: To determine whether the principal source of chemostimuli for GC-D+ neurons is external or internal to the nose, we assessed the consequences of removing external chemostimuli for afferent activity to the necklace glomeruli. To do so, we performed unilateral naris occlusions in Gucy2d Mapt-lacZ +/- mice [which express a beta-galactosidase (beta-gal) reporter specifically in GC-D+ neurons] followed by immunohistochemistry for beta-gal and a glomerular marker of afferent activity, tyrosine hydroxylase (TH). We observed a dramatic decrease in TH immunostaining, consistent with reduced or absent afferent activity, in both necklace and non-necklace glomeruli ipsilateral to the occluded naris. CONCLUSION: Like other MOB glomeruli, necklace glomeruli exhibit a large decrease in afferent activity upon removal of external stimuli. Thus, we conclude that activity in GC-D+ neurons, which specifically innervate necklace glomeruli, is not dependent on internal stimuli. Instead, GC-D+ neurons, like other OSNs in the MOE, primarily sense the external world. PMID- 19284706 TI - Casein haplotypes and their association with milk production traits in Norwegian Red cattle. AB - A high resolution SNP map was constructed for the bovine casein region to identify haplotype structures and study associations with milk traits in Norwegian Red cattle. Our analyses suggest separation of the casein cluster into two haplotype blocks, one consisting of the CSN1S1, CSN2 and CSN1S2 genes and another one consisting of the CSN3 gene. Highly significant associations with both protein and milk yield were found for both single SNPs and haplotypes within the CSN1S1-CSN2-CSN1S2 haplotype block. In contrast, no significant association was found for single SNPs or haplotypes within the CSN3 block. Our results point towards CSN2 and CSN1S2 as the most likely loci harbouring the underlying causative DNA variation. In our study, the most significant results were found for the SNP CSN2_67 with the C allele consistently associated with both higher protein and milk yields. CSN2_67 calls a C to an A substitution at codon 67 in beta-casein gene resulting in histidine replacing proline in the amino acid sequence. This polymorphism determines the protein variants A1/B (CSN2_67 A allele) versus A2/A3 (CSN2_67 C allele). Other studies have suggested that a high consumption of A1/B milk may affect human health by increasing the risk of diabetes and heart diseases. Altogether these results argue for an increase in the frequency of the CSN2_67 C allele or haplotypes containing this allele in the Norwegian Red cattle population by selective breeding. PMID- 19284707 TI - Comparison of classification methods for detecting associations between SNPs and chick mortality. AB - Multi-category classification methods were used to detect SNP-mortality associations in broilers. The objective was to select a subset of whole genome SNPs associated with chick mortality. This was done by categorizing mortality rates and using a filter-wrapper feature selection procedure in each of the classification methods evaluated. Different numbers of categories (2, 3, 4, 5 and 10) and three classification algorithms (naive Bayes classifiers, Bayesian networks and neural networks) were compared, using early and late chick mortality rates in low and high hygiene environments. Evaluation of SNPs selected by each classification method was done by predicted residual sum of squares and a significance test-related metric. A naive Bayes classifier, coupled with discretization into two or three categories generated the SNP subset with greatest predictive ability. Further, an alternative categorization scheme, which used only two extreme portions of the empirical distribution of mortality rates, was considered. This scheme selected SNPs with greater predictive ability than those chosen by the methods described previously. Use of extreme samples seems to enhance the ability of feature selection procedures to select influential SNPs in genetic association studies. PMID- 19284708 TI - Performance comparison of dwarf laying hens segregating for the naked neck gene in temperate and subtropical environments. AB - This study compares laying performances between two environments of dwarf laying hen lines segregating for the naked neck mutation (NA locus), a selected dwarf line of brown-egg layers and its control line. Layers with one of the three genotypes at the NA locus were produced from 11 sires from the control line and 12 sires from the selected line. Two hatches produced 216 adult hens in Taiwan and 297 hens in France. Genetic parameters for laying traits were estimated in each environment and the ranking of sire breeding values was compared between environments. Laying performance was lower, and mortality was higher in Taiwan than in France. The line by environment interaction was highly significant for body weight at 16 weeks, clutch length and egg number, with or without Box-Cox transformation. The selected line was more sensitive to environmental change but in Taiwan it could maintain a higher egg number than the control line. Estimated heritability values in the selected line were higher in France than in Taiwan, but not for all the traits in the control line. The rank correlations between sire breeding values were low within the selected line and slightly higher in the control line. A few sire families showed a good ranking in both environments, suggesting that some families may adapt better to environmental change. PMID- 19284709 TI - Blunt apical dissection during anatomic radical retropubic prostatectomy. AB - BACKGROUND: Meticulous apical dissection during a radical prostatectomy is imperative to achieve desirable pathologic and quality of life outcomes. FINDINGS: We describe a novel technique using careful blunt dissection to better delineate the apex of the prostate, providing a simple means to potentially lessen positive surgical margins at the apex and promote better continence and erectile function in men undergoing an anatomic radical prostatectomy.Median operative time and blood loss were 190 minutes and 675 mL, respectively. Only 10 percent of the patients with positive surgical margins were found to have apical positive surgical margins. Ninety-three percent of patients reported no urinary leakage. CONCLUSION: We believe our technique of isolating the DVC with blunt dissection and then ligating and transecting the DVC to be feasible approach that requires larger studies to truly confirm its utility. PMID- 19284710 TI - Validation of phylogenetic signals in amplified fragment length data: testing the utility and reliability in closely related taxa. AB - BACKGROUND: Discriminating taxa with the nuclear marker, amplified fragment length polymorphism (AFLP) has been accomplished for various organisms in economic, ecological, and evolutionary studies. The protocol available for AFLP generation does not require prior knowledge of the genome; however, it is often extensively modified to fit the needs of the researcher. Modification of this protocol for new labs is intimidating and time-consuming, particularly for taxa in which AFLP have not been previously developed. Furthermore, determining what constitutes quality output during different stages of fragment generation is not well defined and this may further hinder the use AFLP by new researchers. FINDINGS: We present a step-by-step AFLP protocol, using flourophore-labeled primers for use with automated sequencers, including examples of both successful and unsuccessful results. We sufficiently normalized peak intensity and standardized allele calling across all samples for each primer combination. Repeatability was assessed with a phylogenetic tree in which replicate samples clustered together using the minimum evolution procedure. We found differences greater than 10% in allele position among replicated samples would cause replicates to no longer cluster. To minimize offset allele positions, we suggest that researchers analyze different primer combinations at the same time using multiple dyes with the automated sequencer to minimize mismatched alleles across replicates. CONCLUSION: For researchers wanting to use AFLP, this molecular technique is difficult and time-consuming to develop. Clarifying what constitutes quality output for each step in AFLP generation will help to reduce redundant trials in protocol development and, in turn, advance the discipline of population genetics. PMID- 19284711 TI - The protein Nalpha-terminal acetyltransferase hNaa10p (hArd1) is phosphorylated in HEK293 cells. AB - BACKGROUND: The hNaa10p (hArd1) protein is the catalytic subunit of the human NatA Nalpha-terminal acetyltransferase complex. The NatA complex is associated with ribosomes and cotranslationally acetylates human proteins with Ser-, Ala-, Thr-, Val-, and Gly- N-termini after the initial Met- has been removed. In the flexible C-terminal tail of hNaa10p, there are several potential phosphorylation sites that might serve as points of regulation. FINDINGS: Using 2D-gel electrophoresis and hNaa10p specific antibodies, we have investigated whether hNaa10p is phosphorylated in HEK293 cells. Several differently charged forms of hNaa10p are present in HEK293 cells and treatment with Calf Intestine Alkaline Phophatase (CIAP) strongly suggests that hNaa10p is phosphorylated at multiple sites under various cell culture conditions. A direct or indirect role of GSK-3 kinase in regulating hNaa10p phosphorylation is supported by the observed effects of Wortmannin and LiCl, a GSK-3 activator and inhibitor, respectively. CONCLUSION: We demonstrate that hNaa10p protein is phosphorylated in cell culture potentially pointing at phosphorylation as a means of regulating the function of one of the major Nalpha-terminal acetyltransferases in human cells. PMID- 19284713 TI - Poisson analysis of streptococcal bond strengthening on stainless steel with and without a salivary conditioning film. AB - Poisson analysis of retract force-distance curves in atomic force microscopy (AFM) has yielded a new dimension to the decoupling of individual bond forces into a hydrogen bonding and nonspecific force component. Accordingly, bacterial adhesion forces have been decoupled into a hydrogen bonding and nonspecific Lifshitz-Van der Waals contribution. Due to the forced nature of AFM contact, the nonspecific force contribution has hitherto turned out to be repulsive in the analysis of bacterial adhesion forces on nonconducting surfaces. In this study, we present the results of a Poisson analysis of adhesion forces for streptococci adhering to a conducting surface. Adhesion forces measured between stainless steel, both in the absence and presence of an adsorbed salivary conditioning film, increased with increasing contact time between the streptococcal AFM probe and the surface. Concurrent with the increase in adhesion force, there was an increase in the number of minor force peaks in the retract force-distance curves. Poisson analyses of the adhesion forces indicated repulsive nonspecific Lifshitz Van der Waals forces for streptococci adhering to saliva-coated stainless steel, but interestingly and for the first time, attractive nonspecific forces were revealed on stainless steel in the absence of a salivary conditioning film. We tentatively attribute this to attraction between the negatively charged streptococci and their positive image charges in the conducting material, which cannot develop in a nonconducting material or in the presence of a nonconductive protein layer on the stainless steel surface. PMID- 19284712 TI - Verification of the ProPneumo-1 assay for the simultaneous detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae in clinical respiratory specimens. AB - BACKGROUND: Mycoplasma pneumoniae and Chlamydophila pneumoniae are major causes of lower and upper respiratory infections that are difficult to diagnose using conventional methods such as culture. The ProPneumo-1 (Prodesse, Waukesha, WI) assay is a commercial multiplex real-time PCR assay for the simultaneous detection of M. pneumoniae and/or C. pneumoniae DNA in clinical respiratory samples. OBJECTIVE: The aim of this study was to evaluate the sensitivity and specificity of the ProPneumo-1, a newly developed commercial multiplex real-time PCR assay. METHODS: A total of 146 clinical respiratory specimens, collected from 1997 to 2007, suspected of C. pneumoniae or M. pneumoniae infections were tested retrospectively. Nucleic acid was extracted using an automated NucliSense easyMag (bioMerieux, Netherlands). We used a "Home-brew" monoplex real-time assay as the reference method for the analysis of C. pneumoniae and culture as the reference method for the analysis of M. pneumoniae. For discordant analysis specimens were re-tested using another commercial multiplex PCR, the PneumoBacter-1 assay (Seegene, Korea). RESULTS: Following discordant analysis, the sensitivity of the ProPneumo-1 assay for pathogens, C. pneumoniae or M. pneumoniae, was 100%. The specificity of the ProPneumo-1 assay, however, was 100% for C. pneumoniae and 98% for M. pneumoniae. The limits of detection were 1 genome equivalent (Geq) per reaction for pathogens, M. pneumoniae and C. pneumoniae. Due to the multipex format of the ProPneumo-1 assay, we identified 5 additional positive specimens, 2 C. pneumoniae in the M. pneumoniae-negative pool and 3 M. pneumoniae in the C. pneumoniae-negative pool. CONCLUSION: The ProPneumo-1 assay is a rapid, sensitive and effective method for the simultaneous detection of M. pneumoniae and C. pneumoniae directly in respiratory specimens. PMID- 19284714 TI - Highly regioselective synthesis of benz[a]anthracene derivatives via a Pd catalyzed tandem C-H activation/biscyclization reaction. AB - A palladium-catalyzed tandem C-H activation/biscyclization reaction of propargylic carbonates with terminal alkynes was determined, which allowed the tetracyclic benz[a]anthracene framework to be constructed with high regioselectivity. A possible mechanism for this tandem C-H activation/biscyclization process was discussed. PMID- 19284715 TI - Further examination of antiradical properties of Crocus sativus stigmas extract rich in crocins. AB - Data for antiradical properties of saffron extract and its bioactive constituents (crocins, crocetin) are limited and poorly discussed in comparison with those of extracts containing potent scavengers. Further examination was sought using the Folin-Ciocalteu (F-C) reagent and various free radical species produced in cell free or cell model systems. Oregano and turmeric methanol extracts, rich in well established scavengers, and also crocetin, rosmarinic acid, and curcumin, representing the major types of constituents in the three studied extracts, were used as "reference". On the same weight basis, saffron extract activity was found to be rather negligible in all cell-free systems with regard to that found for reference ones. On the contrary, in the human monocyte system, saffron extracts or free crocetin were found to reduce ROS production as effectively as the phenolic antioxidants. Our findings point out that saffron extracts exhibit a remarkable intracellular antioxidant activity that cannot be revealed using assays repeatedly applied to the evaluation of phenolic-type antioxidants. PMID- 19284716 TI - Gas/particle partitioning of two acid-base active compounds in mainstream tobacco smoke: nicotine and ammonia. AB - Gas/particle (G/P) partitioning constant (Kp) values are reported for nicotine and ammonia for mainstream tobacco smoke (MTS) for a selection of cigarettes, "little cigars", and biddies. As K(p)(nic) decreases as a result of the increasing basicity in the MTS, there is an increase in volatility of nicotine from the smoke particulate matter. The "little cigars" and biddies exhibited generally lower K(p)(nic) values and higher unbound ammonia levels than most of the cigarettes, suggesting a correlation between the two parameters. However, within just the cigarettes, there was little correlation. The water content of MTS particulate matter was found to affect both K(p)(nic) and K(p)(amm). Unbound ammonia is actual NH3/NH4+; bound ammonia is comprised of compounds such as amides of ammonia; total ammonia is unbound + bound. Most historical studies of ammonia in MTS have not accurately measured either unbound or total ammonia: the acidic solutions historically employed to determine ammonia in MTS will release ammonia from bound forms by hydrolysis, and the release in those studies may not have been complete. This study concludes that a thorough examination of unbound and bound ammonia in MTS will be required before the role of ammonia in affecting volatility of nicotine in MTS can be understood. PMID- 19284717 TI - An enantioselective organocatalytic approach to both enantiomers of lasubine II. AB - A concise stereoselective route providing access to both enantiomers of the bioactive quinolizidine alkaloid lasubine II has been developed. The enantioselectivity was introduced by taking advantage of a proline-catalyzed asymmetric Mannich reaction. Next, the bicyclic system was constructed via a diastereoselective Mannich cyclization and subsequent ring-closing metathesis as the key steps. PMID- 19284718 TI - Selective 5-hydroxytryptamine 2C receptor agonists derived from the lead compound tranylcypromine: identification of drugs with antidepressant-like action. AB - We report here the design, synthesis, and pharmacological properties of a series of compounds related to tranylcypromine (9), which itself was discovered as a lead compound in a high-throughput screening campaign. Starting from 9, which shows modest activity as a 5-HT(2C) agonist, a series of 1-aminomethyl-2 phenylcyclopropanes was investigated as 5-HT(2C) agonists through iterative structural modifications. Key pharmacophore feature of this new class of ligands is a 2-aminomethyl-trans-cyclopropyl side chain attached to a substituted benzene ring. Among the tested compounds, several were potent and efficacious 5-HT(2C) receptor agonists with selectivity over both 5-HT(2A) and 5-HT(2B) receptors in functional assays. The most promising compound is 37, with 120- and 14-fold selectivity over 5-HT(2A) and 5-HT(2B), respectively (EC(50) = 585, 65, and 4.8 nM at the 2A, 2B, and 2C subtypes, respectively). In animal studies, compound 37 (10-60 mg/kg) decreased immobility time in the mouse forced swim test. PMID- 19284719 TI - Ionic recognition by 7-nitro-1,3,5-triaza adamantane: first thermodynamic study. AB - A thermodynamic study involving 7-nitro-1,3,5-triaza adamantane, 1, and its interaction with metal cations in nonaqueous media is first reported. Solubility data of 1 in various solvents were used to derive the standard Gibbs energies of solution, DeltaG(s)o in these solvents. The effect of solvation in the different media was assessed from the Gibbs energy of transfer taking acetonitrile as a reference solvent. 1H NMR studies of the interaction of 1 and metal cations were carried out in CD3CN and CD3OD and the data are reported. Conductance measurements revealed that this ligand forms lead(II) or zinc complexes of 1:1 stoichiometry in acetonitrile. It also revealed a stoichiometry of two molecules of 1 per mercury(II) and two cadmium (II) ions per molecule of 1. The addition of silver salt to 1 led to the precipitation of the silver-1 complex which was isolated and characterized by X-ray crystallography. At variance with conductance measurements in solution, in the solid state the X-ray structure shows a 1:1 stoichiometry in the Hg(II) complex. The thermodynamics of complexation of 1 and these cations provide a quantitative assessment of the selective behavior of this ligand for ions of environmental relevance. PMID- 19284720 TI - Effect of electrodes on electronic transport of molecular electronic devices. AB - Understanding the effects of each component of a molecular device is at the heart of designing a useful device. Molecular cores and linkers are well studied, but relatively few studies have been devoted to investigating the electrode effect on a molecular electronic device. Here, we study unique characteristics of Au, Ru, and carbon nanotube electrodes using the nonequilibrium Green function method combined with a density functional theory. By systematic modification of the device region, we extract the effect of the electrode materials on the electron transport. We show that the band structure and surface density of states of an electrode material, independent of the choice of other device components, have unique influences on the transmission curve. We note that carbon nanotube electrodes can offer unusual nonlinear current-voltage characteristics. PMID- 19284721 TI - Three-body dissociative charge exchange dynamics of sym-triazine. AB - Translational spectroscopy and coincidence detection techniques have been employed to observe the three-body dissociation dynamics resulting from charge exchange between a fast molecular beam of the sym-triazine cation (C(3)N(3)H(3)(+)) and cesium. The energetics and momentum correlation between the three hydrogen cyanide products suggest that dissociation proceeds via two mechanisms that reflect the initial excited states in which sym-triazine is produced. Monte Carlo simulations support a stepwise mechanism resulting from initial excitation into the 3s Rydberg state. Excitation into the valence pi* <-- n manifold was found to proceed via a near-symmetric concerted mechanism, in agreement with recent theoretical studies on the dissociation of sym-triazine. PMID- 19284722 TI - Photodissociation dynamics of alkyl nitrites at 266 and 355 nm: the OH product channel. AB - Photodissociation of methyl nitrite and n-butyl nitrite at 266 and 355 nm has been investigated in the gas phase at room temperature. OH photoproducts were observed, and their internal state distributions were measured by the one-photon laser-induced fluorescence (LIF) technique. It was found that the nascent OH from the 266 nm photolysis of methyl nitrite was vibrationally cold, and its rotational state distribution conformed to a Boltzmann behavior with a rotational temperature of T(rot) = 2200 +/- 150 K. In contrast, the nascent OH from the 266 nm photolysis of n-butyl nitrite was found to be vibrationally excited, and the measured relative population of v'' = 0:1 was 0.78:0.22. The rotational state distribution of the OH v'' = 1 state conformed to Boltzmann behavior, with a rotational temperature of T(rot) = 1462 +/- 120 K. However, a simple Boltzmann distribution was not found for the OH v'' = 0 state. In the photolysis of n-butyl nitrite at 355 nm, the OH fragment was found to be vibrationally cold and its rotational state distribution showed non-Boltzmann behavior. A photodissociation mechanism involving an intramolecular hydrogen atom transfer process is proposed for the OH product pathway for methyl nitrite, which has been compared with the potential energy surfaces obtained from density functional theory (DFT) calculations. A photodissociation mechanism of n-butyl nitrite is also proposed for the OH product pathway, which differs from that of methyl nitrite due to the effects of the different alkoxy substituents. PMID- 19284723 TI - Benzothiazines in synthesis. Further studies of the intramolecular, stereoselective addition of sulfonimidoyl carbanions to alpha,beta-unsaturated functional groups. AB - A variety of alkenes substituted by electron-withdrawing groups serve as competent electrophiles for the stereoselective, intramolecular nucleophilic addition of sulfonimidoyl carbanions to form benzothiazines. This reaction generally proceeds with complete stereoselectivity within the limits of our detection. In some cases, benzothiazine formation occurs in a single pot at relatively high temperatures during N-arylation of the simple sulfoximine used in this study. Yet, the process occurs with the same direction and extent of stereoselectivity as that seen when the Michael addition is performed at very low temperatures. PMID- 19284724 TI - Variations in conjugated linoleic acid (CLA) content of processed cheese by lactation time, feeding regimen, and ripening. AB - Dairy products are major sources of conjugated linoleic acid (CLA); thus, an increase in CLA content can improve the quality value of dairy products. The objective of this work was to determine the effects of lactation time, feeding regimen, and ripening period on the level of CLA in processed cheese. CLA content in milk varied with the period of lactation; high in spring (April and May, about 6.8 mg CLA/g fat) and relatively low in mid summer and winter (about 4.3 mg CLA/g fat). The effects of dietary regimen and ripening period were determined in milk, which was obtained from March to May. After aging for 4 months, the cheese made from milk obtained from cows fed on pasture contained relatively higher levels of CLA compared to cheese made from milk obtained from cows fed indoors (8.12 mg CLA/g fat vs 6.76 mg CLA/g fat), but there was no difference in 7 month-aged cheeses. In both pasture and indoor feeding, 7 month-aged cheeses showed higher CLA content than 4 month-aged cheeses. The contents of stearic acid (C18:0) and linolenic acid (C18:3) were significantly higher in cheese from pasture fed cows compared to those in cows fed indoors. These findings should be helpful for the efficient production of functional dairy products with high CLA contents. PMID- 19284725 TI - Maturity stage at harvest determines the fruit quality and antioxidant potential after storage of sweet cherry cultivars. AB - Eleven sweet cherry cultivars were harvested at three maturity stages (S1 to S3) based on skin color and stored at 2 degrees C for 16 days and a further period of 2 days at 20 degrees C (shelf life, SL) to analyze quality (color, total soluble solids, and total acidity) and bioactive compounds (total phenolics and anthocyanins) and their relationship to total antioxidant activity (TAA), determined in hydrophilic (H-TAA) or lipophilic (L-TAA) fraction. For all cultivars and maturity stages, the ripening process advanced during postharvest storage with increases in color intensity and decreases in acidity, as well as enhancements in phenolics, anthocyanins, and TAA in both H-TAA and L-TAA, although important differences existed among cultivars. The results showed that sweet cherry should be harvested at stage S3 (4 days later than the commercial harvest date) since after 16 days of cold storage + SL, the highest antioxidant capacity was achieved for both H-TAA and L-TAA. PMID- 19284726 TI - Self-assembly of Janus cylinders into hierarchical superstructures. AB - We present in-depth studies of the size tunability and the self-assembly behavior of Janus cylinders possessing a phase segregation into two hemicylinders. The cylinders are prepared by cross-linking the lamella-cylinder morphology of a polystyrene-block-polybutadiene-block-poly(methyl methacrylate) block terpolymer. The length of the Janus cylinders can be adjusted by both the amplitude and the duration of a sonication treatment from the micro- to the nanometer length. The corona segregation into a biphasic particle is evidenced by selective staining of the PS domains with RuO(4) and subsequent imaging. The self-assembly behavior of these facial amphiphiles on different length scales is investigated combining dynamic light scattering (DLS), small-angle neutron scattering (SANS), and imaging procedures. Cryogenic transmission electron microscopy images of the Janus cylinders in THF, which is a good solvent for both blocks, exhibit unimolecularly dissolved Janus cylinders with a core-corona structure. These results are corroborated by SANS measurements. Supramolecular aggregation takes place in acetone, which is a nonsolvent for polystyrene, leading to the observation of fiber-like aggregates. The length of these fibers depends on the concentration of the solution. A critical aggregation concentration is found, under which unimolecularly dissolved Janus cylinders exist. The fibers are composed of 2-4 Janus cylinders, shielding the inner insoluble polystyrene hemicylinder against the solvent. Herein, the SANS data reveal a core-shell structure of the aggregates. Upon deposition of the Janus cylinders from more concentrated solution, a second type of superstructure is formed on a significantly larger length scale. The Janus cylinders form fibrillar networks, in which the pore size depends on the concentration and deposition time of the sample. PMID- 19284727 TI - Spectroscopy by integration of frequency and time domain information for fast acquisition of high-resolution dark spectra. AB - A simple and effective method, SIFT (spectroscopy by integration of frequency and time domain information), is introduced for processing nonuniformly sampled multidimensional NMR data. Applying the computationally efficient Gerchberg Papoulis (G-P) algorithm, used previously in picture processing and medical imaging, SIFT supplements data at nonuniform points in the time domain with the information carried by known "dark" points (i.e., empty regions) in the frequency domain. We demonstrate that this rapid integration not only removes the severe pseudonoise characteristic of the Fourier transforms of nonuniformly sampled data, but also provides a robust procedure for using frequency information to replace time measurements. The latter can be used to avoid unnecessary sampling in sampling-limited experiments, and the former can be used to take advantage of the ability of nonuniformly sampled data to minimize trade-offs between the signal-to-noise ratio and the resolution in sensitivity-limited experiments. Processing 2D and 3D data sets takes about 0.1 and 2 min, respectively, on a personal computer. With these several attractive features, SIFT offers a novel, model-independent, flexible, and user-friendly tool for efficient and accurate processing of multidimensional NMR data. PMID- 19284728 TI - Multiwalled carbon nanotubes specifically inhibit osteoclast differentiation and function. AB - Since attention has been paid to the use of multiwalled carbon nanotubes (MWCNTs) as biomaterials in contact with bone, it is critical to understand the reaction of bone cells to MWCNTs. We show that MWCNTs inhibit osteoclastic bone resorption in vivo and that MWCNTs inhibit osteoclastic differentiation and suppressed a transcription factor essential for osteoclastogenesis in vitro. These results suggest that MWCNTs have beneficial effects on bones when they are used as biomaterials. PMID- 19284729 TI - Random telegraphic conductance fluctuation at Au-pentacene-Au nanojunctions. AB - Random telegraphic noises with a preferential jump height around 0.018G(0) were observed in the conductance traces of the Au-pentacene-Au junctions. The distribution of the plateau lifetime obeyed an exponential distribution, but the decay constants were independent of the height of the adjacent conductance jumps or the measurement temperature. This result, along with the buckyferrocene (Fe(C(60)(CH(3))(5))C(5)H(5)) result, suggests that the conductance fluctuation originates from the current-induced geometrical fluctuation around a single molecule. PMID- 19284730 TI - "Cloning" of single-walled carbon nanotubes via open-end growth mechanism. AB - Using the concept of "cloning", we report herein a rational approach to grow single-walled carbon nanotubes (SWNTs) with controlled chirality via an open-end growth mechanism. Specifically, by using open-end SWNTs as "seeds/catalysts" (without metal catalysts), "new/duplicate" SWNTs could be grown and cloned from the parent segments via an open-end growth mechanism. Using this strategy, we have measured more than 600 short seed segments and have found that the yield of cloning is relatively low (around 9%). This yield can be greatly improved up to 40% by growing SWNTs on quartz substrate. Atomic force microscopy and micro resonance Raman spectroscopy characterization indicate that the parent nanotube and the duplicate nanotube have the same structure. These findings provide a potential approach for growing SWNTs with controlled chirality, which are important for the application of SWNTs in nanoelectronics. PMID- 19284731 TI - Self-assembled nanogaps via seed-mediated growth of end-to-end linked gold nanorods. AB - Gold nanorods (AuNRs) are of interest for a wide range of applications, ranging from imaging to molecular electronics, and they have been studied extensively for the past decade. An important issue in AuNR applications is the ability to self assemble the rods in predictable structures on the nanoscale. We here present a new way to end-to-end link AuNRs with a single or few linker molecules. Whereas methods reported in the literature so far rely on modification of the AuNRs after the synthesis, we here dimerize gold nanoparticle seeds with a water-soluble dithiol-functionalized polyethylene glycol linker and expose the linked seeds to growth conditions identical to the synthesis of unlinked AuNRs. Doing so, we obtain a large fraction of end-to-end linked rods, and transmission electron microscopy provides evidence of a 1-2 nm wide gap between the AuNRs. Flow linear dichroism demonstrates that a large fraction of the rods are flexible around the hinging molecule in solution, as expected for a molecularly linked nanogap. By using excess of gold nanoparticles relative to the linking dithiol molecule, this method can provide a high probability that a single molecule is connecting the two rods. In essence, our methods hence demonstrate the fabrication of a nanostructure with a molecule connected to two nanoelectrodes by bottom-up chemical assembly. PMID- 19284732 TI - Recent advances in high oxidation state Mo and W imido alkylidene chemistry. PMID- 19284733 TI - An analysis of the different behavior of DNA and RNA through the study of the mutual relationship between stacking and hydrogen bonding. AB - The mutual relationship between stacking and hydrogen bonding and the possible influence of stacking in the different behavior of DNA and RNA base pairs have been studied through complete DFT optimization of different structures of A-U and A-T dimers (i.e., A-U/U-A and A-T/T-A), using some functionals developed by the group of Truhlar. The results obtained in this work clearly show that stacking and hydrogen bonding are deeply connected. The different behavior of DNA and RNA when replacing uracil by thymine can be interpreted through the formation of a stabilizing CH/pi interaction between the methyl group of thymine and the five member ring of adenine. PMID- 19284734 TI - Label-free DNA sensor based on surface charge modulated ionic conductance. AB - The surface charge effect in controlling ionic conductance through a nanoporous alumina membrane is investigated for its application in a convenient detection method of unlabeled DNA. To this goal, surface modification with mixtures of neutral silanes and morpholinos (neutral analogues of DNA) was optimized to yield a strong effect on ionic conductance change upon DNA binding, which can exceed an order of magnitude. The effect can be employed in fabrication of inexpensive DNA sensors. PMID- 19284735 TI - Total synthesis of aigialomycin D using a Ramberg-Backlund/RCM strategy. AB - The bioactive resorcylic acid lactone aigialomycin D (1) has been synthesized by a novel combination of ring-closing metathesis (RCM) and Ramberg-Backlund reactions. This synthetic strategy enables the C1'-C2' alkene to be masked as a sulfone during formation of the macrocycle by ring closing metathesis at the C7' C8' olefin, thus avoiding competing formation of a cyclohexene. A subsequent Ramberg-Backlund reaction efficiently produces the C1'-C2' E-alkene. This combined RCM/Ramberg-Backlund reaction strategy should be widely applicable to the synthesis of macrocyclic dienes. PMID- 19284736 TI - The remarkable cis effect in the ene reactions of nitrosocarbonyl intermediates. AB - Nitrosocarbonyls are fleeting and highly reactive intermediates that undergo ene reactions in a two-step fashion. The addition steps are rate and product determining and lead to polarized diradicals that readily enter the H-abstraction step yielding the ene products. The addition TSs are reached early, and the stabilizing CH...O contacts drive the reactions to the cis adducts. B3LYP calculations alone do not describe the correct ordering of addition TSs in the ene reaction with trimethylethylene and (E)- and (Z)-3-methyl-2-pentenes. Only at the MPWB1K level of treatment, medium-range noncovalent interactions are successfully recovered, accounting satisfactorily for the experimental selectivities. The more stable and isolable ArNOs exhibit late addition TSs, and distortion energies become dominant driving the reaction exclusively to the Markovnikov adducts. PMID- 19284737 TI - Rhodium-catalyzed vinylcyclopropanation/cyclopentenation of strained alkenes via a sequential carborhodation process. AB - A rhodium-catalyzed reaction of dienylboronate esters with alkenes is described. Strained bicyclic alkenes show the highest reactivity toward the rhodium catalyzed addition of the dienylboronate esters. Depending on the substitution pattern of dienylboronate esters, an intramolecular 1,6- or 1,4-addition mechanism may be operative, affording carbocycles containing a vinylcyclopropane or cyclopentene moiety. PMID- 19284738 TI - Enzymatic desymmetrization of prochiral 2-substituted-1,3-diamines: preparation of valuable nitrogenated compounds. AB - A wide range of prochiral 1,3-diamines were first efficiently synthesized and subsequently desymmetrized by using lipase from Pseudomonas cepacia as catalyst and diallyl carbonate as alkoxycarbonylating agent. In all cases, the amino carbamates of R-configuration were recovered. Final selective cleavage of the N allyloxycarbonyl moiety was carried out under mild reaction conditions, which demonstrates the high versatility and potential of this chemoenzymatic route as a source of intermediates in the synthesis of related optically active nitrogenated derivatives. PMID- 19284739 TI - PhIO as a powerful cyclizing reagent: regiospecific [3+2]-tandem oxidative cyclization of imine toward cofacially self-aggregated low molecular mass organic materials. AB - The powerful cyclization and tandem oxidation property of environmentally benign PhIO is developed for the first time, which leads to regiospecific [3+2]-tandem oxidative cyclization of imine at room temperature in rapid access to a new class of compounds, 1,2-functionalized 4,5-diarylimidazoles, in excellent yield with synthetic efficiency. Size, shape, and activity of the involved nanoreactors for the green approach built up from various surfactants are investigated. Spontaneous generation of low molecular mass self-aggregated organic materials, their cofacial one-dimensional packing, and interesting photophysical properties are reported. PMID- 19284740 TI - Synthesis of a fluorine-substituted puromycin derivative for Bronsted studies of ribosomal-catalyzed peptide bond formation. AB - The mechanism by which the ribosome catalyzes peptide bond formation remains controversial. Here we describe the synthesis of a nucleoside that can be used in Bronsted experiments to assess the transition state of ribosome catalyzed peptide bond formation. This substrate is the nucleoside 3'-amino-3'-deoxy-3'-[(3''R)-3 fluoro-l-phenyl-alanyl]-N(6),N(6)-dimethyladenosine, which was prepared from (1R,2R)-2-amino-1-phenylpropane-1,3-diol. This substrate is active in peptide bond formation on the ribosome and is a useful probe for Bronsted analysis experiments on the ribosome. PMID- 19284741 TI - Mechanisms of antagonism of the GluR2 AMPA receptor: structure and dynamics of the complex of two willardiine antagonists with the glutamate binding domain. AB - Ionotropic glutamate receptors mediate the majority of vertebrate excitatory synaptic transmission. The development of selective antagonists for glutamate receptor subtypes is of interest in the treatment of a variety of neurological disorders. This study presents the crystal structure of the binding domain of GluR2 bound to two antagonists (UBP277 and UBP282) that are derivatives of the natural product, willardiine. The antagonists bind to one lobe of the protein with interactions similar to agonists. Interaction with the second lobe differs between the two antagonists, resulting in a different position of the uracil ring and different orientations of the bilobed structure. UBP277 binding produces a stable lobe orientation that is similar to the apo state, but the binding of UBP282 produces the largest hyperextension of the lobes yet reported for an AMPA receptor. The carboxyethyl (UBP277) and carboxybenzyl (UBP282) substituents in the N(3) position keep the lobes separated by a "foot-in-the-door" mechanism and the internal dynamics are minimal compared to the CNQX-bound form of the protein (which makes minimal contacts with one of the two lobes). In contrast to the antagonists CNQX and DNQX, UBP277 and UBP282 produce complexes with higher thermal stability, but affinities that are more than 100-fold lower. These structures support the idea that antagonism is associated with the overall orientation of the lobes rather than with specific interactions, and antagonism can rise either from specific interactions with both lobes ("foot-in-the-door" mechanism) or from the lack of extensive interactions with one of the two lobes. PMID- 19284743 TI - Antioxidant hydroquinones substituted by beta-1,6-linked oligosaccharides in wheat germ. AB - Seven new compounds that demonstrate antioxidant properties, 4-hydroxy-3 methoxyphenyl beta-d-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranoside (1), 4-hydroxyphenyl beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranoside (2), 4-hydroxy-3-methoxyphenyl beta D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl-(1- >6)beta-D-glucopyranoside (3), 4-hydroxy-3-methoxyphenyl beta-D-glucopyranosyl-(1 ->6)-beta-D-glucopyranoside (4), 4-hydroxy-3,5-dimethoxyphenyl beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranoside (5), 4-hydroxy-3,5-dimethoxyphenyl beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranoside (6), and 4-hydroxy-2-methoxyphenyl beta-d-glucopyranosyl-(1-->6) beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside (7), were isolated from wheat germ. The structures were determined by spectroscopic and chemical methods. Compound 1 was the most abundant, approximately 2 mg isolated from each gram of wheat germ. The antioxidant activity of compounds 1-7 was determined by the Trolox equivalent antioxidant capacity assay, and 2 and 7 showed higher values than the other compounds. Compounds 1 and 3-6 reacted with the radical cation reagent within a few seconds, whereas 2 and 7 required several minutes for complete reaction. Compound 1 was shown to protect plasmid DNA from oxidative stress damage caused by hydrogen peroxide; this effect was concentration dependent. PMID- 19284744 TI - Intermolecular vibrational motions of solute molecules confined in nonpolar domains of ionic liquids. AB - In this study, we address the following question about the dynamics of solute molecules in ionic liquids (ILs). Are the intermolecular vibrational motions of nonpolar molecules confined in the nonpolar domains formed by tail aggregation in ILs the same as those in an alkane solvent? To address this question, the optical Kerr effect (OKE) spectrum of CS(2) in the IL 1-pentyl-3-methylimidazolium bis(trifluoromethanesulfonyl)amide ([C(5)mim][NTf(2)]) was studied as a function of concentration at 295 K by the use of optical heterodyne-detected Raman-induced Kerr effect spectroscopy. The OKE spectrum broadens and shifts to higher frequency as the CS(2) concentration is decreased from 20 to 10 mol %; at lower concentrations, no further change in the width of the OKE spectrum is observed. Multicomponent line shape analysis of the OKE spectrum of 5 mol % CS(2) in [C(5)mim][NTf(2)] reveals that the CS(2) and [C(5)mim][NTf(2)] contributions to the spectrum are separable and that the CS(2) contribution is similar to the OKE spectrum of 5 mol % CS(2) in n-pentane with the spectrum being lower in frequency and narrower than that of neat CS(2). These results suggest that, at this concentration, CS(2) molecules are isolated from each other and mainly localized in the nonpolar domains of the IL. PMID- 19284745 TI - Hirseins A and B, daphnane diterpenoids from Thymelaea hirsuta that inhibit melanogenesis in B16 melanoma cells. AB - Two new daphnane diterpenoids, hirseins A (1) and B (2), were isolated from the aerial parts of Thymelaea hirsuta, and their structures were elucidated on the basis of spectroscopic data interpretation. Hirsein B (2) is an unusual daphnane in possessing a coumaroyl moiety. NOESY correlations of 2 implied that isomerization of the coumaroyl group in 2 was caused by equilibrium between the E (2e) and Z (2z) forms. Compounds 1 and 2 were found to inhibit melanogenesis in B16 murine melanoma cells. PMID- 19284746 TI - Gaussian-mixture umbrella sampling. AB - We introduce the Gaussian-mixture umbrella sampling method (GAMUS) , a biased molecular dynamics technique based on adaptive umbrella sampling that efficiently escapes free energy minima in multidimensional problems. The prior simulation data are reweighted with a maximum likelihood formulation, and the new approximate probability density is fit to a Gaussian-mixture model, augmented by information about the unsampled areas. The method can be used to identify free energy minima in multidimensional reaction coordinates. To illustrate GAMUS , we apply it to the alanine dipeptide (2D reaction coordinate) and tripeptide (4D reaction coordinate). PMID- 19284747 TI - Bonding and adhesion at the SiC/Fe interface. AB - Ceramics such as SiC have the potential to act as protective coatings, primarily due to their high melting points and wear resistance. We use periodic density functional theory (DFT) within the generalized gradient approximation (GGA) to calculate the adhesion strength between SiC and Fe, for Si- and C-terminations of SiC(100) and two surfaces of Fe: (100) and (110). We predict a maximum ideal work of adhesion of 6.51 J/m(2) at the SiC(100)/Fe(110) interface for C-Fe interfacial bonding, which is stronger than the traditional chrome coating's adherence to Fe. We characterize the interfacial bonding via local densities of states and electron density difference analysis and find strong covalent bonding and some evidence of metallic bonding between Si (C) and Fe. Our results suggest that SiC might prove useful as a thin adhesion layer in a multilayer protective coating for steel. PMID- 19284748 TI - C-terminal hemocyanin from hemocytes of Penaeus vannamei interacts with ERK1/2 and undergoes serine phosphorylation. AB - To understand molecular immune response of Penaeus vannamei during Taura syndrome virus (TSV) infection, expression and functional proteomics studies were performed on hemocyanin, which is a major abundant protein in shrimp hemocytes. Two-dimensional electrophoresis (2-DE) revealed up-regulation of several C terminal fragments of hemocyanin, whereas the N-terminal fragments were down regulated during TSV infection. 2-D Western blot analysis showed that the C terminal hemocyanin fragments had more acidic isoelectric points (pI), whereas the N-terminal fragments had less acidic pI. Further analysis by NetPhos showed a greater number of serine phosphorylation sites in the C-terminal hemocyanin. Additionally, motif scan using Scansite revealed ERK D-domain, which is required for activation of ERK1/2 effector kinase, as a kinase-binding site at the 527th valine in the C-terminal hemocyanin, whereas neither motif nor functional domain was found in the N-terminus. Co-immunoprecipitation confirmed the interaction between the C-terminal hemocyanin and ERK1/2. 1-D Western blot analysis showed that ERK1/2 was also up-regulated during TSV infection. Our findings demonstrate for the first time that ERK1/2 signaling pathway may play an important role in molecular immune response of P. vannamei upon TSV infection through its interaction with the C-terminal hemocyanin. PMID- 19284749 TI - Novel 2- and 4-substituted 1H-imidazo[4,5-c]quinolin-4-amine derivatives as allosteric modulators of the A3 adenosine receptor. AB - 4-Arylamino and 2- cycloalkyl (including amino substitution) modifications were made in a series of 1H-imidazo[4,5-c]quinolin-4-amine derivatives as allosteric modulators of the human A(3) adenosine receptor (AR). In addition to allosteric modulation of the maximum functional efficacy (in [(35)S]GTPgammaS G protein binding assay) of the A(3)AR agonist Cl-IB-MECA (15), some analogues also weakly inhibited equilibrium radioligand binding at ARs. 4-(3,5-Dichlorophenylamino) (6) or 2-(1-adamantyl) (20) substitution produced allosteric enhancement (twice the maximal agonist efficacy), with minimal inhibition of orthosteric AR binding. 2 (4-Tetrahydropyranyl) substitution abolished allosteric enhancement but preserved inhibition of orthosteric binding. Introduction of nitrogen in the six-membered ring at the 2 position, to improve aqueous solubility and provide a derivatization site, greatly reduced the allosteric enhancement. 2-(4 (Benzoylamino)cyclohexyl) analogues 23 and 24 were weak negative A(3)AR modulators. Thus, consistent with previous findings, the allosteric and orthosteric inhibitory A(3)AR effects in imidazoquinolines are structurally separable, suggesting the possible design of additional derivatives with enhanced positive or negative allosteric A(3)AR activity and improved selectivity in comparison to inhibition of orthosteric binding. PMID- 19284751 TI - Synthesis, antimalarial activity, and preclinical pharmacology of a novel series of 4'-fluoro and 4'-chloro analogues of amodiaquine. Identification of a suitable "back-up" compound for N-tert-butyl isoquine. AB - On the basis of a mechanistic understanding of the toxicity of the 4 aminoquinoline amodiaquine (1b), three series of amodiaquine analogues have been prepared where the 4-aminophenol "metabolic alert" has been modified by replacement of the 4'-hydroxy group with a hydrogen, fluorine, or chlorine atom. Following antimalarial assessment and studies on mechanism of action, two candidates were selected for detailed ADME studies and in vitro and in vivo toxicological assessment. 4'-Fluoro-N-tert-butylamodiaquine (2k) was subsequently identified as a candidate for further development studies based on potent activity versus chloroquine-sensitive and resistant parasites, moderate to excellent oral bioavailability, low toxicity in in vitro studies, and an acceptable safety profile. PMID- 19284753 TI - Tissue proteomics and metabolomics: an excellent start and a promising future. PMID- 19284752 TI - Evaluation of (64)Cu(DO3A-xy-TPEP) as a potential PET radiotracer for monitoring tumor multidrug resistance. AB - In this study, we evaluated the potential of (64)Cu(DO3A-xy-TPEP) (DO3A-xy-TPEP = (2-(diphenylphosphoryl)ethyl)diphenyl(4-((4,7,10-tris(carboxymethyl)-1,4,7,10 tetraazacyclododecan-1-yl)methyl)benzyl)phosphonium) as a PET (positron emission tomography) radiotracer for noninvasive monitoring of multidrug resistance (MDR) transport function in several xenografted tumor models (MDR-negative: U87MG; MDR positive: MDA-MB-435, MDA-MB-231, KB-3-1, and KB-v-1). It was found that (64)Cu(DO3A-xy-TPEP) has a high initial tumor uptake (5.27 +/- 1.2%ID/g at 5 min p.i.) and shows a steady uptake increase between 30 and 120 min p.i. (2.09 +/- 0.53 and 3.35 +/- 1.27%ID/g at 30 and 120 min p.i., respectively) in the MDR negative U87MG glioma tumors. (64)Cu(DO3A-xy-TPEP) has a greater uptake difference between U87MG glioma and MDR-positive tumors (MDA-MB-231: 1.57 +/- 0.04, 1.00 +/- 0.17, and 0.93 +/- 0.15; MDA-MB-435: 1.15 +/- 0.19, 1.12 +/- 0.20, and 0.81 +/- 0.11; KB-3-1: 1.45 +/- 0.31, 1.43 +/- 0.16, and 1.08 +/- 0.19; and KB-v-1: 1.63 +/- 0.47, 1.81 +/- 0.31, and 1.14 +/- 0.22%ID/g at 30, 60, and 120 min p.i., respectively) than (99m)Tc-Sestamibi. Regardless of the source of MDR, the overall net effect is the rapid efflux of (64)Cu(DO3A-xy-TPEP) from tumor cells, which leads to a significant reduction of its tumor uptake. It was concluded that (64)Cu(DO3A-xy-TPEP) is more efficient than (99m)Tc-Sestamibi as the substrate for MDR P-glycoproteins (MDR Pgps) and multidrug resistance associated proteins (MRPs), and might be a more efficient radiotracer for noninvasive monitoring of the tumor MDR transport function. (64)Cu(DO3A-xy-TPEP) and (99m)Tc-Sestamibi share almost identical subcellular distribution patterns in U87MG glioma tumors. Thus, it is reasonable to believe that (64)Cu(DO3A-xy-TPEP), like (99m)Tc-Sestamibi, is able to localize in mitochondria due to the increased plasma and mitochondrial transmembrane potentials in tumor cells. PMID- 19284754 TI - Enzymatic solubilization of brewers' spent grain by combined action of carbohydrases and peptidases. AB - Brewers' spent grain (BSG), a high-volume coproduct from the brewing industry, primarily contains proteins, barley cell wall carbohydrates, and lignin. To create new possibilities for the exploitation of this large biomass stream, the solubilization of BSG by the combined action of carbohydrases (Depol 740 and Econase) and peptidase (Alcalase and Promod 439) was explored. Hydrolysis protocols were optimized with respect to temperature (influencing both microbial contamination and rate of enzymatic hydrolysis), pH, enzyme dose, order of enzyme addition, and processing time. On the basis of this approach, one- and two-step protocols are proposed taking 4-8 h and yielding combined or separate fractions of hydrolyzed oligosaccharides and liberated hydrolyzed protein. Optimized procedures resulted in the solubilization of >80% of the proteinaceous material, up to 39% of the total carbohydrates, and up to 42% of total dry matter in BSG. Of the original xylan present in BSG, 36% could be solubilized. Sequential and simultaneous treatments with the two enzyme types gave similar results. In sequential processes, the order of the carbohydrase and peptidase treatments had only minor effects on the outcome. Depol 740 released more pentoses than Econase and gave slightly higher overall dry matter solubilization yields. PMID- 19284755 TI - Controllable epitaxial crystallization and reversible oriented patterning of two dimensional colloidal crystals. AB - We demonstrate a reliable and highly efficient epitaxial templating approach for the formation of two-dimensional (2D) colloidal crystals. By applying an alternating electric field (AEF), one-dimensional colloidal lines are used as an epitaxial template to site specifically initiate 2D colloidal crystallization and control the orientation of the 2D colloidal crystals. The kinetics of the crystallization and structure ordering is precisely and conveniently manipulated by the external AEF. The well-defined artificial linear defects are embedded inside the 2D colloidal crystals by means of heteroepitaxy, whereas the unwanted existing defects are controllably relaxed via an electrically induced annealing process. This epitaxial templating approach is fast, reversible, and amenable for large-area oriented patterning of colloidal crystals, providing a new way for the creation of novel materials and devices with functions and properties that can be reversibly changed, such as electrically tunable photonic waveguides and e papers. PMID- 19284756 TI - Dualistic nature of the mechanism of the Morita-Baylis-Hillman reaction probed by electrospray ionization mass spectrometry. AB - The Morita-Baylis-Hillman (MBH) reaction allows chemists to form new sigma C-C bonds in a single-step straightforward manner and thus to construct densely functionalized molecules for further chemical manipulation. Using electrospray ionization for transferring ions directly from solution to the gas phase, and mass (and tandem mass) spectrometry for mass and structural assignments, new key intermediates for the rate-determining step of the MBH reaction have been successfully intercepted and structurally characterized. These ESI-MS data provide experimental evidence supporting recent suggestions, based on kinetic experiments and theoretical calculations, for the dualist nature of the proton transfer step of the MBH mechanism. PMID- 19284757 TI - Collective dynamics of protein hydration water by brillouin neutron spectroscopy. AB - By a detailed experimental study of THz dynamics in the ribonuclease protein, we could detect the propagation of coherent collective density fluctuations within the protein hydration shell. The emerging picture indicates the presence of both a dispersing mode, traveling with a speed greater than 3000 m/s, and a nondispersing one, characterized by an almost constant energy of 6-7 meV. In agreement with molecular dynamics simulations [Phys. Rev. Lett. 2002, 89, 275501], the features of the dispersion curves closely resemble those observed in pure liquid water [Phys. Rev. E: Stat. Phys., Plasmas, Fluids, Relat. Interdiscip. Top. 2004, 69, 061203]. On the contrary, the observed damping factors are much larger than in bulk water, with the dispersing mode becoming overdamped at Q = 0.6 A(-1) already. Such novel experimental findings are discussed as a dynamic signature of the disordering effect induced by the protein surface on the local structure of water. PMID- 19284758 TI - Alkyne-linked 2,2-disubstituted-indolin-3-one oligomers as extended beta-strand mimetics. AB - The design, synthesis, and structural analysis of a new foldamer that mimics an extended beta-sheet are presented. The nonpeptidic mimetic is based on a series of 2,2-disubstituted-indolin-3-one groups linked through their 4,7-positions by an alkyne spacer. An intramolecular hydrogen bond between the carbonyl of one indolinone subunit and the proximal NH of another subunit imposes a conformation that mimics the side chain positioning on a beta-strand. X-ray crystallographic studies support the presence of this intramolecular hydrogen bond. The described approach allows extension of the scaffold to longer oligomers that will form the basis of new mimetics for the disruption of therapeutically relevant protein protein interactions that rely on the contacts of side chain residues on two beta strands. PMID- 19284759 TI - Optimizing the extraction of phenolic antioxidants from peanut skins using response surface methodology. AB - Peanut skins are a byproduct of peanut blanching operations and contain high levels of phenolic antioxidants. The effect of solvent type (methanol MeOH, ethanol EtOH, and water), concentration (0, 30, 60, 90%), temperature (30, 45, 60 degrees C), and time (10, 20, 30 min) on total phenolic content (TPC), oxygen radical absorbance capacity (ORAC) level, and resveratrol content of peanut skins was investigated. Response surface methodology was used to estimate the optimum extraction conditions for each solvent. EtOH extracts had the highest TPC followed by MeOH and water. The maximum predicted TPC under the optimized conditions (30.8%, 30.9 degrees C, 12 min) was 118 mg of gallic acid equivalents (GAE)/g of skins. MeOH extracts had the highest ORAC activity of 2149 micromol of TE/g followed by EtOH and water under the optimized conditions of 30% MeOH, 52.9 degrees C and 30 min. Resveratrol was identified in MeOH extracts but was not found in samples extracted with EtOH or water. PMID- 19284760 TI - A tetranuclear Cu(II)-based 2D aggregate with an unprecedented tetradentate mu(4) N1,N3,N7,N9-adeninate nucleobase. AB - A unique tetranuclear Cu(II)-based 2D aggregate containing 26-membered Cu(II)ade macrocycles, [Cu(4)(DMF)(6)(mu(4)-ade)(2)(mu(2)-Cl)(2)Cl(4)](n) (1; Hade = adenine, DMF = N,N-dimethylformamide), was isolated by the reaction of CuCl(2) and Hade in a mixed DMF-methanol medium and structurally characterized by X-ray crystallography, elemental analysis, Fourier transform infrared, fluorescence spectroscopy, and thermogravimetry-differential thermal analysis techniques. Unexpectedly, the adeninate in 1 represents an unprecedented tetradentate mu(4) N1,N3,N7,N9-bridging mode, which significantly contributes to both the aggregate of four Cu(II) cores within the subunit and the extension of the 2D covalent framework. Additionally, 1 displays the intense Hade-based fluorescence emission in solution at room temperature. PMID- 19284761 TI - A heterobimetallic gallyl complex containing an unsupported Ga-Y bond. AB - The synthesis and characterization of the first unsupported Ga-Y bond in [Y{Ga(NArCH)(2)}{C(PPh(2)NSiMe(3))(2)}(THF)(2)] (Ar = 2,6-diisopropylphenyl) is described; structural and computational analyses are consistent with a highly polarized covalent Ga-Y bond. PMID- 19284762 TI - Polynuclear spin crossover complexes: synthesis, structure, and magnetic behavior of [Fe4(mu-CN)4(phen)4(L)2)]4+ squares. AB - Three new tetranuclear compounds of formula [Fe(4)(mu CN)(4)(phen)(4)(L)(2))](PF(6))(4) x G where L = tris(pyridin-2-ylmethyl)amine (TPMA) [G = 0] (1), (6-methylpyrid-2-ylmethyl)-bis(pyrid-2-ylmethyl)amine (MeTPMA) [G = 0] (2), or bis(6-methylpyrid-2-ylmethyl)-(pyrid-2-ylmethyl)amine (Me(2)TPMA) [G = NH(4)PF(6)] (3) and phen = 1,10-phenanthroline) have been synthesized and characterized. The three compounds crystallize in the C2/c space group and consist of [Fe(4)(mu-CN)(4)(phen)(4)(L)(2))](4+) square shaped cations with two distinct iron(II) sites. The sites, associated with [Fe(phen)(2)(CN)(2)] and [Fe(L)(NC)(2)] moieties, are connected by cyanide bridging ligands and reside in different [FeN(4)C(2)] and [FeN(6)] ligand field strength environments. For 1, the structural features of both sites at 100 and 293 K are those of an iron(II) atom in the low-spin state, according to the magnetic properties. At 370 K the structure of the [FeN(6)] site is consistent with a quite complete change of spin state from the low-spin state to the high-spin state, a behavior confirmed by the magnetic study. Introduction of a methyl substituent in the sixth position of one or two pyridine groups to get MeTPMA or Me(2)TPMA derivatives, respectively, induce in 2 and 3 notable steric constraint in the [FeN(6)] site making longer the average Fe-N bond distances thereby weakening the ligand field strength and stabilizing the high-spin state. The [FeN(4)C(2)] site remains in the low-spin state in the three compounds. PMID- 19284763 TI - Regioselective catenation of dinuclear palladium and platinum metallocycles promoted by pi...pi interactions. AB - Dinuclear metallocycles were assembled from an L-shaped bidentate ligand based on 4,4'-bipyridine and palladium or platinum square-planar cis complexes. The palladium (4a,b) or platinum (5a,b) square metallocycles were obtained as 1:1 regioisomeric mixtures depending on which pair of coordinative nitrogen atoms binds to the metal center. The squares display a pi-deficient cavity suitable to incorporate two pi-donor aromatic systems. Therefore, the reaction with macrocyclic polyethers (BPP34C10 or DN38C10) resulted in the regioselective self assembly of the [3]catenanes 4a(BPP34C10 or DN38C10)(2) x 6 PF(6) and 5a(BPP34C10 or DN38C10)(2) x 6 PF(6) because only macrocycles 4a and 5a present the correct disposition of the pi-deficient aromatic systems to maximize the pi...pi stacking interactions. Single-crystal X-ray analyses of [3]catenanes revealed that the structures are additionally stabilized by [C-H...O], [N-H...O] bonds and [C H...pi] interactions. The 1:2 inclusion complexes of metallocycles were prepared by self-assembly of three components: the ligand 1-(4-(pyridin-4-yl)benzyl)-4,4' bipyridin-1-ium, a square planar complex M(en)(NO(3))(2) (M = Pd or Pt, en = ethylenediamine), and a dioxoaromatic guest in a 2:2:2 ratio. The comparative study of the formation of 1:2 inclusion complexes has allowed us to conclude that the pi...pi interactions between the host and guests are responsible for the observed regioselectivity. PMID- 19284764 TI - Phenanthrene-based tylophorine-1 (PBT-1) inhibits lung cancer cell growth through the Akt and NF-kappaB pathways. AB - Tylophorine and related natural compounds exhibit potent antitumor activities. We previously showed that PBT-1, a synthetic C9-substituted phenanthrene-based tylophorine (PBT) derivative, significantly inhibits growth of various cancer cells. In this study, we further explored the mechanisms and potential of PBT-1 as an anticancer agent. PBT-1 dose-dependently suppressed colony formation and induced cell cycle G2/M arrest and apoptosis. DNA microarray and pathway analysis showed that PBT-1 activated the apoptosis pathway and mitogen-activated protein kinase signaling. In contrast, PBT-1 suppressed the nuclear factor kappaB (NF kappaB) pathway and focal adhesion. We further confirmed that PBT-1 suppressed Akt activation accelerated RelA degradation via IkappaB kinase-alpha and down regulated NF-kappaB target gene expression. The reciprocal recruitment of RelA and RelB on COX-2 promoter region led to down-regulation of transcriptional activity. We conclude that PBT-1 induces cell cycle G2/M arrest and apoptosis by inactivating Akt and by inhibiting the NF-kappaB signaling pathway. PBT-1 may be a good drug candidate for anticancer chemotherapy. PMID- 19284766 TI - Synthesis, optical, and mesomorphic properties of self-assembled organogels featuring phenylethynyl framework with elaborated long-chain pyridine-2,6 dicarboxamides. AB - A series of new organogelators with pi-conjugated phenylethynyl framework featuring long-chain carboxamides have been synthesized. These organgelators have shown great ability to gel a variety of organic solvents to form stable organogels with the minimum gelation concentration as low as 0.1 wt %. Gelation is completely thermoreversible, and it occurs due to the aggregation of the organogelators resulting in the formation of a fibrous network via a combination of intermolecular hydrogen bonding, pi-pi stacking, and van der Waals interactions that is observed for the xerogels by TEM. The influence of sol-to gel transition has been explored in detail by variable-temperature 1H NMR, UV-vis absorption, and fluorescence spectroscopy. Aggregation-induced enhanced emission has been observed in these organogelator molecules with an order of higher fluorescence quantum yields from solution to gels. In addition, some molecules also exhibit unique liquid crystalline properties over a large temperature range as revealed by DSC and POM studies. PMID- 19284767 TI - Cholesterol-appended aromatic imine organogelators: a case study of gelation driven component selection. AB - This letter describes a novel approach for developing organogelators through the formation of reversible imine bonds from two molecular components and the enriching behavior of the gelating imines. Cholesterol-appended aniline 1 and 4 substituted benzaldehydes 2a-d did not gelate any solvents. Their condensation products, imines 3a-d, however, could gelate alcohols because of the enhanced stacking interaction of the imine unit. For a further component selectivity test, the reactions of the mixture of 1, 2b-d, and cholesterol-free aniline 7 (1:1:1) in different solvents were performed. The resulting imines were reduced to the corresponding amines and analyzed with 1H NMR. It was revealed that, for the reactions resulting in no formation of the gel phase, imines 8a-c formed from 2b d and 7 were obtained as the major product (64-76%) and all of the reactions that led to the formation of the gel phase gave rise to 3b-d as the major product (55 61%). PMID- 19284768 TI - Organogels based on 1H-imidazolecarboxamide amphiphiles. AB - We prepared 1H-imidazolecarboxamide amphiphiles as potential organogelators. Compounds A1-A3, in which an imidazole head was connected to a hydrophobic trialkyloxyphenyl group, showed an ability to gelate nonpolar solvents, including alkanes. The dry gels obtained from compounds A1-A3 had columnar hexagonal structures. Polycatenar 1H-imidazolecarboxamide amphiphile B2, consisting of a 1H imidazole head connected through a benzene ring to a tridecyloxyphenyl tail, formed an organogel in DMSO. In a concentrated THF solution (30 wt %), compound B2 exhibited a lyotropic liquid-crystalline phase with a columnar hexagonal structure. X-ray diffraction (XRD) results suggested a molecular arrangement consisting of a disk, via hydrogen bonding between successive imidazole moieties, and an assembly of columnar structures. PMID- 19284769 TI - Self-assembled fibers photopolymerized in nematic liquid crystals: stable electrooptical switching in light-scattering mode. AB - Liquid-crystalline (LC) gels showing stable light-scattering electrooptical properties have been prepared by the photopolymerization of a gelator forming self-assembled fibers in a room-temperature nematic liquid crystal. A polymerizable gelator having methacryloyl moieties self-assembles into finely dispersed fibers in the nematic liquid crystal, resulting in the formation of LC physical gels. Photopolymerization of the gelator in the gel states leads to the preparation of cross-linked fibrous networks. The LC polydomain structures effective for light-scattering electrooptical switching are kept after polymerization. The polymerized LC gels show stable reversible switching of electrooptical responses with enhanced threshold properties between 0 and 70 V. PMID- 19284770 TI - Anions as efficient chain stoppers for hydrogen-bonded supramolecular polymers. AB - The chain length of hydrogen-bonded supramolecular polymers and thus their rheological properties can be controlled by the presence of so-called chain stoppers: these monofunctional monomers are able to interact with the monomers and break the polymer chains. In this letter, we show that the use of anions, strong hydrogen bond competitors, instead of precisely designed complementary units is a very simple approach to tuning the rheology of a bisurea-based hydrogen-bonded supramolecular polymer. All of the anions tested were able to break the supramolecular chains, resulting in a dramatic drop in the viscosity of the solutions and were found to be more efficient than a previously described organic stopper. A careful study of the rheological properties of bisurea solutions in the presence of H2PO4,N(C4H9)4 showed that the presence of this ion does not modify the nature of the bisurea supramolecular assembly. For a molar fraction of stopper of only 10(-5), the viscosity of bisurea solutions decreases by a factor of 10 as a result of the formation of shorter supramolecular assemblies. PMID- 19284771 TI - Two-component organogelators based on two L-amino acids: effect of combination of L-lysine with various L-amino acids on organogelation behavior. AB - A two-component organogelation system, consisting of Nepsilon-dodecyl-L-lysine esters (amine components) and N-dodecyl-L-amino acids (valine, phenylalanine, alanine, glycine, L-lysine) as acid components, was proposed, and its organogelation properties were investigated. The mixture of amine and acid components produced an organic salt compound through acid-base interactions, and it created a three-dimensional network by entangling self-assembled nanofibers through hydrogen bonding and van der Waals interactions, thus leading to the formation of organogels. The combination of Lys/Phe yielded rigid dodecane gel, and the gelators of Lys/ Lys yielded a thermally stable dodecane gel. The organogelation properties significantly depended on the combination of amine and acid components, and they could be controlled by the selection of suitable components. PMID- 19284772 TI - Ugi-post functionalization, from a single set of ugi-adducts to two distinct heterocycles by microwave-assisted palladium-catalyzed cyclizations: tuning the reaction pathways by ligand switch. AB - Linear amides 4, prepared in one step by the Ugi four-component reaction, were converted to 3,4-dihydroquinoxalin-3-ones (5) or to 2-(2-oxoindolin-1 yl)acetamides (6) dependent on the catalytic conditions. While microwave irradiation was found to be determinant on the reaction efficiency, the choice of ligand diverged the reaction pathways. Heating a solution of 4 in dioxane/MeCN (v/v = 85/15) under microwave irradiation conditions in the presence of Pd(dba)(2) (0.05 equiv) and Cs(2)CO(3) (2 equiv), using XPhos as a supporting ligand, afforded the 3,4-dihydroquinoxalin-3-ones (5) via an intramolecular N arylation of the secondary amide. On the other hand, using BINAP as ligand under otherwise identical conditions, intramolecular alpha-CH arylation of tertiary amide occurred to furnish the oxindoles (6). PMID- 19284773 TI - Donator acceptor map of psittacofulvins and anthocyanins: are they good antioxidant substances? AB - Psittacofulvins represent an unusual class of pigments (noncarotenoid lipochromes), which are found only in the red, orange, and yellow plumage of parrots. Anthocyanins are flavonoids, and they are one of the primary types of colorants found in plants. Blue butterflies acquire blue and UV hues on their wings, owing to the presence of flavonoids. It is assumed that these natural pigments are valuable antioxidants because they are able to scavenge free radicals. The aim of this investigation is to rationalize the scavenging activity of psittacofulvins and anthocyanins, in terms of the one electron transfer mechanism, taking into account that to prevent oxidative stress, substances must either donate or accept electrons. Density functional approximation calculations are used to obtain ionization potentials, electron affinities, electrodonating, and electroaccepting power indexes. Taking these values, a donator acceptor map (DAM) was constructed, indicating that anthocyanins are good electron donors, whereas psittacofulvins are good electron acceptors. Anthocyanins and vitamins are antioxidants, whereas psittacofulvins and carotenoids are antireductants (oxidants). In terms of solvent effects, animal pigments (carotenoids, psittacofulvins, and anthocyanins) are much better electron acceptors in water than in either the gas phase or benzene. Solvent effects do not alter the electron donor capacity of vitamins, but anthocyanins become effective electron acceptors in water, rather than effective electron donors. The information presented here may also be valuable for the design and analysis of further experiments. PMID- 19284774 TI - Locally optimized coordinates in modified Shepard interpolation. AB - An extension of the modified Shepard interpolation method is presented that allows expansions for the potential energy using different local coordinate sets to be used in a global interpolation. The coordinates used in a given Taylor expansion are determined using a training set of geometries at which the ab initio potential energy is known and that is built up during the construction of the interpolated potential energy surface. The method is applied to the bound state potential energy surface of methanol and a significant improvement in the rate of convergence of the interpolated potential energy surface to the ab initio potential energy is observed. PMID- 19284775 TI - Controlling the retention of small molecules in emulsion microdroplets for use in cell-based assays. AB - Water-in-oil microdroplets in microfluidics are well-defined individual picoliter reaction compartments and, as such, have great potential for quantitative high throughput biological screening. This, however, depends upon contents of the droplets not leaking out into the oil phase. To assess the mechanism of possible leaking, the retention of various fluorescein derivatives from droplets formed in mineral oil and stored for hours in a reservoir on chip was studied. Leaking into the oil phase was observed and was shown to be dependent on the nature of the compounds and on the concentration of the silicone-based polymeric surfactant Abil EM 90 used. In experiments in which droplets filled with fluorescein were mixed with droplets filled with only buffer, the rate of efflux from filled droplets to empty droplets was dependent on the number of neighboring droplets of different composition. Buffer droplets with five fluorescein-containing neighbors took up the fluorophore 4.5 times faster than buffer droplets without fluorescein neighbors. The addition of bovine serum albumin (BSA) substantially reduced leaking. A formulation with 5% BSA reduces leaking of the fluorophore from 45% to 3%. Inclusion of BSA enabled experiments to be carried out over periods up to 18 h, without substantial leaking (<5%). We demonstrate the utility of this additive by following the enzymatic activity of alkaline phosphatase expressed by Escherichia coli cells. The ability to reliably compartmentalize genotype (cell) and phenotype (reaction product) is the basis for using microdroplets in directed evolution studies, and the approaches described herein provide a test system for assessing emulsion formulations for such purposes. PMID- 19284777 TI - Fully automatic sample treatment by integration of microextraction by packed sorbents into commercial capillary electrophoresis-mass spectrometry equipment: application to the determination of fluoroquinolones in urine. AB - This paper describes a new and innovative way to integrate microextraction by packed sorbents (MEPS) into commercial CE equipment. The suggested integration allows the automatic sample cleanup and preconcentration requiring only a few microliters of sample and no additional hardware and software. The MEPS was integrated in the outlet region of a commercial CE equipment cartridge in order to provide easy manipulation and exchange. The robustness of the proposed integration was demonstrated by the design and use of a (MEPS)-nonaqueous capillary electrophoresis (NACE)-MS method used to determine fluoroquinolones "FQs" (namely, ofloxacin, marbofloxacin, enrofloxacin, danofloxacin, and difloxacin) in urine. The method allows the analysis of micrograms per liter of FQs to be carried out with only 48 microL of urine sample. The obtained LODs were in the range 6.3-10.6 microg/L. An analysis of spiked urine samples was used to validate the method. Absolute recoveries were in the range of 71-109% while the precision expressed as repetitivity of peak area was lower than 5.9%. PMID- 19284776 TI - Plasmonic nanoholes in a multichannel microarray format for parallel kinetic assays and differential sensing. AB - We present nanohole arrays in a gold film integrated with a six-channel microfluidic chip for parallel measurements of molecular binding kinetics. Surface plasmon resonance effects in the nanohole arrays enable real-time, label free measurements of molecular binding events in each channel, while adjacent negative reference channels can record measurement artifacts such as bulk solution index changes, temperature variations, or changing light absorption in the liquid. With the use of this platform, streptavidin-biotin specific binding kinetics are measured at various concentrations with negative controls. A high density microarray of 252 biosensing pixels is also demonstrated with a packing density of 10(6) sensing elements/cm(2), which can potentially be coupled with a massively parallel array of microfluidic channels for protein microarray applications. PMID- 19284778 TI - Fragment-based discovery of nonpeptidic BACE-1 inhibitors using tethering. AB - BACE-1 (beta-site amyloid precursor protein cleaving enzyme), a prominent target in Alzheimer's disease drug discovery efforts, was surveyed using Tethering technology to discover small molecule fragment ligands that bind to the enzyme active site. Screens of a library of >15000 thiol-containing fragments versus a panel of BACE-1 active site cysteine mutants under redox-controlled conditions revealed several novel amine-containing fragments that could be selectively captured by subsets of the tethering sites. For one such hit class, defined by a central aminobenzylpiperidine (ABP) moiety, X-ray crystal structures of BACE mutant-disulfide conjugates revealed that the fragment bound by engaging both catalytic aspartates with hydrogen bonds. The affinities of ABP fragments were improved by structure-guided chemistry, first for conjugation as thiol-containing fragments and then for stand-alone, noncovalent inhibition of wild-type (WT) BACE 1 activity. Crystallography confirmed that the inhibitors bound in exactly the same mode as the disulfide-conjugated fragments that were originally selected from the screen. The ABP ligands represent a new type of nonpeptidic BACE-1 inhibitor motif that has not been described in the aspartyl protease literature and may serve as a starting point for the development of BACE-1-directed Alzheimer's disease therapeutics. PMID- 19284779 TI - Ultrathin nanofibrous films prepared from cadmium hydroxide nanostrands and anionic surfactants. AB - We developed a simple fabrication method of ultrathin nanofibrous films from the dispersion of cadmium hydroxide nanostrands and anionic surfactants. The nanostrands were prepared in a dilute aqueous solution of cadmium chloride by using 2-aminoethanol. They were highly positively charged and gave bundlelike fibers upon mixing an aqueous solution of anionic surfactant. The nanostrand/surfactant composite fibers were filtered on an inorganic membrane filter. The resultant nanofibrous film was very uniform in the area of a few centimeters square when the thickness was not less than 60 nm. The films obtained with sodium tetradecyl sulfate (STS) had a composition close to the electroneutral complex, [Cd37(OH)68(H2O)n] x 6(STS), as confirmed by energy dispersive X-ray analysis. They were water-repellent with a contact angle of 117 degrees, and the value slightly decreased with the alkyl chain length of anionic surfactants. Ultrathin nanofibrous films were stable enough to be used for ultrafiltration at pressure difference of 90 kPa. We could effectively separate Au nanoparticles of 40 nm at an extremely high filtration rate of 14000 L/(h m2 bar). PMID- 19284780 TI - A convenient, one-step synthesis of benzyl (Ar) ureas of the type ArCH(2)NHCONHR from Ar and R(1)OCH(2)NHCONHR via ureidoalkylation. AB - A method for the one-step C-ureidoalkylation of phenol, anisole, or aniline rings furnishing ArCH(2)NHCONHR (Ar = benzyl) products in moderate to good yields is described. With phenol ring systems, higher yields were attained when the reaction was worked up with an acidic ethanethiol addition to cleave any O ureidoalkylation products that formed during the reaction. PMID- 19284781 TI - Angle dependent collective surface plasmon resonance in an array of silver nanoparticles. AB - Theoretical analysis of the scattering efficiency of an equidistantly spaced regular array of spherical silver nanoparticles reveals a nonmonotonic shift of the collective SPR wavelength and its bandwidth depending on the distance between the particles and the angle of the incidence of the linear polarized electromagnetic wave. The far-field electromagnetic coupling between the particles in the chain exhibits the largest range of angular tuning of the collective SPR band when the distance between the particles in the chain approaches that of the collective SPR wavelength. The dependence of the SPR wavelength and its bandwidth on the angle of the incidence of the linear polarized electromagnetic wave and the distance between the particles in the chain provides an additional flexibility for the development of optical biochemical sensors and subwavelength waveguides. PMID- 19284782 TI - Mass spectrometry-based structural dissection of fluorescent proteins. AB - Fluorescent proteins (FPs) are essential for live cell studies using fluorescence microscopy. To date, the molecular basis for FPs' irreversible photobleaching and the nature of the associated photoproducts are a matter of debate. Mass spectrometry, which should be an ideal technique for the structural dissection of FPs, cannot be harnessed efficiently due to their extreme resistance to trypsinolysis, due to the compactness of the barrel structure containing the chromopeptide. We devised a mild endoproteolysis procedure that affords a peptide mass fingerprint almost totally covering the sequence, thus allowing high resolution mass spectrometric investigations of the protein structure. PMID- 19284783 TI - Immunologic glycosphingolipidomics and NKT cell development in mouse thymus. AB - Invariant NKT cells are a hybrid cell type of Natural Killer cells and T cells, whose development is dependent on thymic positive selection mediated by double positive thymocytes through their recognition of natural ligands presented by CD1d, a nonpolymorphic, non-MHC, MHC-like antigen presenting molecule. Genetic evidence suggested that beta-glucosylceramide derived glycosphingolipids (GSLs) are natural ligands for NKT cells. N-butyldeoxygalactonojirimycin (NB-DGJ), a drug that specifically inhibits the glucosylceramide synthase, inhibits the endogenous ligands for NKT cells. Furthermore, we and others have found a beta linked glycosphingolipid, isoglobotriaosylceramide (iGb3), is a stimulatory NKT ligand. The iGb3 synthase knockout mice have a normal NKT development and function, indicating that other ligands exist and remain to be identified. In this study, we have performed a glycosphingolipidomics study of mouse thymus, and studied mice mutants which are deficient in beta-hexosaminidase b or alpha galactosidase A, two glycosidases that are up- and downstream agents of iGb3 turnover, respectively. Our mass spectrometry methods generated a first database for glycosphingolipids expressed in mouse thymus, which are specifically regulated by rate-limiting glycosidases. Among the identified thymic glycosphingolipids, only iGb3 is a stimulatory ligand for NKT cells, suggesting that large-scale fractionation, enrichment and characterization of minor species of glycosphingolipids are necessary for identifying additional ligands for NKT cells. Our results also provide early insights into cellular lipidomics studies, with a specific focus on the important immunological functions of glycosphingolipids. PMID- 19284784 TI - Approaching solid tumor heterogeneity on a cellular basis by tissue proteomics using laser capture microdissection and biological mass spectrometry. AB - The purpose of this study was to examine solid tumor heterogeneity on a cellular basis using tissue proteomics that relies on a functional relationship between Laser Capture Microdissection (LCM) and biological mass spectrometry (MS). With the use of LCM, homogeneous regions of cells exhibiting uniform histology were isolated and captured from fresh frozen tissue specimens, which were obtained from a human lymph node containing breast carcinoma metastasis. Six specimens approximately 50,000 cell each (three from tumor proper and three from tumor stroma) were collected by LCM. Specimens were processed directly on LCM caps, using sonication in buffered methanol to lyse captured cells, solubilize, and digest extracted proteins. Prepared samples were analyzed by LC/MS/MS resulting in more than 500 unique protein identifications. Decoy database searching revealed a false-positive rate between 5 and 10%. Subcellular localization analysis for stromal cells revealed plasma membrane 14%, cytoplasm 39%, nucleus 11%, extracellular space 27%, and unknown 9%; and tumor cell results were 5%, 58%, 26%, 4%, and 7%, respectively. Western blot analysis confirmed specific linkage of validated proteins to underlying pathology and their potential role in solid tumor heterogeneity. With continued research and optimization of this method including analysis of additional clinical specimens, this approach may lead to an improved understanding of tumor heterogeneity, and serve as a platform for solid tumor biomarker discovery. PMID- 19284785 TI - Systematic LC-MS analysis of labile post-translational modifications in complex mixtures. AB - Most proteins are post-translationally modified and the characterization of modified peptides in complex mixtures generated by enzymatic digestion of multiple proteins remains a major analytical challenge. We describe an integrated LC-MS workflow implemented on a hybrid quadrupole time-of-flight (Q-ToF) instrument to detect modified peptides in a complex peptide sample and establish the nature of the modification. The method is based on the alternating acquisition of full mass spectra under different collision conditions inducing the cleavage of the substituents. Modified peptides are detected based on their specific fragmentation generating the nonmodified peptide backbone and reporter ions in the low mass region. The two mass analyzer stages of a Q-ToF instrument are used to eliminate the low mass chemical background in the quadrupole and thus facilitate the detection of low mass reporter ions in the ToF. Off-line data processing enables detection of one (or even multiple) modifications and the modified candidates are subsequently sequenced in a directed MS/MS mode. The technique was applied to the analysis of O-GlcNAc peptides, a very complex mixture of N-linked glycopeptides, and a phosphotyrosine peptide. PMID- 19284786 TI - Serum proteomic approach for the identification of serum biomarkers contributed by oral squamous cell carcinoma and host tissue microenvironment. AB - The lack of serum biomarkers for head and neck carcinoma limits early diagnosis, monitoring of advanced disease, and prediction of relapses in patients. We conducted a comprehensive proteomics study on serum from mice bearing orthotopic human oral squamous cell carcinomas (OSCC) with distinct invasive phenotypes. Matched established cell lines were transplanted orthotopically into tongues of RAG-2/gamma(c) mice and mouse serum was analyzed by 2-dimensional-differential gel electrophoresis(2D-DIGE)/liquid chromatography (LC)-MS/MS and by online 2D-LC MS/MS of iTRAQ labeled samples. We identified several serum proteins as being differentially expressed between control and cancer-bearing mice and between noninvasive and invasive cancer (p<0.05). Differentially expressed proteins of human origin included the epidermal growth factor receptor (EGFR), cytokeratins, G-protein coupled receptor 87, Rab11 GTPase, PDZ-domain containing proteins, and PEST-containing nuclear proteins. Identified proteins of mouse origin included clusterin, titin, vitronectin, vitamin D-binding protein, hemopexin, and kininogen I. The levels of serum and cell secreted EGFR were further validated to match proteomic data regarding the inverse correlation with the invasive phenotype. In summary, we report a comprehensive patient-based proteomics approach for the identification of potential serum biomarkers for OSCC using an orthotopic xenograft mouse model. PMID- 19284787 TI - Alpha(E)C, the C-terminal extension of fibrinogen, has chaperone-like activity. AB - Human fibrinogen is an important coagulation factor as well as an acute phase protein in the circulatory system. Fibrinogen-420 is distinguished from the conventional alpha chain of fibrinogen-340 by the presence of an additional 236 residue carboxyl terminus globular domain (alpha(E)C). The alpha(E)C domain of human fibrinogen-420 is a stable and early proteolytic cleavage product in the circulation. A genuine physiological function for alpha(E)C has not yet been established. Our study aims to characterize the novel chaperone-like activity of alpha(E)C. alpha(E)C efficiently protects a series of model proteins from thermally induced aggregation. Furthermore, alpha(E)C specifically recognizes the partially denatured form instead of the native form of citrate synthase (CS) and potentially protects it from thermally induced inactivation. The protective effect may result from formation of soluble complexes between alpha(E)C and partially denatured CS as tested by size exclusion column and electron microscope. In addition, alpha(E)C can keep the partially denatured luciferase in a folding competent state and help it refold in cooperation with rabbit reticulocyte lysate (RRL). Furthermore, alpha(E)C can also form complexes with thermally stressed plasma proteins. Our findings reveal the novel function of alpha(E)C as a chaperone-like protein, which not only provides new insights into the extracellular chaperone system but also has implications on the physiological and pathological relevance of fibrinogen. PMID- 19284790 TI - Optically modulated fluorophores for selective fluorescence signal recovery. AB - Fluorescence imaging in biological sciences is hindered by significant depth dependent signal attenuation and highly fluorescent backgrounds. We have developed optically modulated near-IR-emitting few-atom Ag nanodots that are selectively and dynamically photobrightened upon simultaneous excitation with a secondary laser, enabling high-sensitivity image extraction to reveal only the demodulated fluorophores. Image demodulation is demonstrated in high-background environments to extract weak signals from completely obscuring background emission. PMID- 19284791 TI - Pest toxicology: the primary mechanisms of pesticide action. AB - Pesticides are used to control pests before they harm us or our crops. They are selective toxicants in the form and manner used. Pesticides must be effective without human or crop injury. They must also be safe relative to human and environmental toxicology. The study of how the pesticide works on the pest is referred to here as pest toxicology. About 700 pesticides, including insecticides, herbicides, and fungicides, act on perhaps 95 biochemical targets in pest insects, weeds, and destructive fungi. Current insecticides act primarily on four nerve targets, i.e., acetylcholinesterase, the voltage-gated chloride channel, the acetylcholine receptor, and the gamma-aminobutyric acid receptor, systems which are present in animals but not plants. Herbicides act mostly on plant specific pathways by blocking photosynthesis, carotenoid synthesis, or aromatic and branched chain amino acid synthesis essential in plants but not mammals. Many fungicides block ergosterol (the fungal sterol) or tubulin biosynthesis or cytochrome c reductase, while others disrupt basic cellular functions. A major limiting factor in the continuing use of almost all pesticides is the selection of strains not only resistant to the selecting or pressuring compounds but also cross-resistant to other pesticides acting at the same target. One approach to reinstating control is to shift from compounds with the resistant target site or mode of action to another set which have a sensitive target. This type of pesticide management led to the formation of Resistance Action Committees for insecticides, herbicides, and fungicides with very knowledgable experts to define resistance groups, which are in fact listings of primary target sites in pest toxicology. Continued success in pest and pesticide management requires an understanding of comparative biochemistry and molecular toxicology considering pests, people, and crops. Defining and applying the principles of pest toxicology are critical to food production and human health. PMID- 19284792 TI - Color tuning via adjusting the dye-loading capacity of a polymer. AB - Color tuning from monomer emission to aggregate emission was accomplished by vacuum thermal evaporation of N,N'-di(n-butyl)quinacridone onto UV-cured photopolymer (NOA-63) with different exposure time. The diffusion of dye molecules into the polymer matrix can be controlled by varying the curing time, which induces the change in the mole ratio between the monomers that are molecularly dispersed into polymer and the aggregates that are shielded on the polymer surface. On the basis of this principle, dual-color and multicolor luminescence patterning were demonstrated by depositing dye molecules onto photolithographically patterned NOA-63 with binary masks and grayscale masks, respectively. PMID- 19284793 TI - How do colloidal aggregates yield to compressive stress? AB - Aqueous dispersions of silica nanoparticles have been aggregated through the addition of Al13 polycations and then submitted to osmotic compression. The structures of these dispersions have been determined through small-angle neutron scattering, before and after compression. Some dispersions consisted of mixtures of aggregated and nonaggregated particles--actually a few aggregates dispersed in a "sea" of nonaggregated particles. In these dispersions, it was found that the resistance to osmotic compression originated from the ionic repulsions of the nonaggregated particles; the compression law that related the applied osmotic pressure Pi to the silica volume fraction Phi was Pi approximately [Phi/(1 Phi)]2. Other dispersions were fully aggregated, with all particles forming a fractal network that extended throughout the available volume. In these dispersions, it was found that the resistance to compression originated from surface-surface interparticle bonds. The application of low osmotic pressures (<50 kPa) resulted in compression at macroscopic scales only (>300 nm), while the structure of the network at local and mesoscopic scales was unchanged. Accordingly, few interparticle bonds were broken, and the deformation was primarily elastic. The compression law for this elastic deformation was in agreement with the predicted scaling law Pi approximately Phi4. The application of higher osmotic pressures (>50 kPa) resulted in compression at macroscopic and mesoscopic scales (30-300 nm), while the local structure was still retained. Accordingly, many more interparticle bonds were broken. The compression law for this plastic deformation was in agreement with a scaling prediction of Pi approximately Phi1.7. The location of the elastic-plastic transition indicated that the strength of the interparticle bonds was on the order of 5 times the thermal energies at ambient temperature. PMID- 19284794 TI - Synthesis of hexahelicene and 1-methoxyhexahelicene via cycloisomerization of biphenylyl-naphthalene derivatives. AB - The new approach provides nonphotochemical syntheses of helicenes based on the easy, convergent, and modular assembly of key biphenylyl-naphthalenes and their platinum-catalyzed double cycloisomerization. This sequence of reactions provides a synthetic route to helicenes in two steps from simply accessible building blocks. Furthermore, the method enables the introduction of substituents into the hexahelicene skeleton. The strategy developed is exemplified by the synthesis of 6,10-dimethylhexahelicene and 1-methoxy-6,10-dimethylhexahelicene. PMID- 19284795 TI - Theoretical study of dioxygen binding process in iron(III) catechol dioxygenase: "oxygen activation" vs "substrate activation". AB - Dioxygen binding process of nonheme iron(III) center in intradiol catechol dioxygenase was investigated with CASSCF/CASPT2 method to incorporate multiconfigurational character participating in Fe-O(2) interaction. In this process, two alternative mechanisms were proposed: one is called "oxygen activation" and the other is called "substrate activation". Our CASSCF/CASPT2 calculated results support the oxygen activation. Potential energy curves and electronic structure evaluated with SA(state-averaged)-CASSCF/CASPT2 method indicate that the charge transfer directly occurs from the catecholate moiety to the dioxygen moiety in the O(2) binding process, to produce eta(1)-end-on type iron(III)-superoxo complex. This is the key step of the dioxygen activation. Interestingly, the iron center always keeps high spin d(5) character during the O(2) binding process, indicating the iron(III) center does not receive charge transfer from the catecholate moiety. However, this does not mean that the iron(III) center is not necessary to the dioxygen activation. The important role which the iron(III) center plays in catechol dioxygenase is to adjust the energy level of O(2) to induce the charge transfer from the catecholate moiety to the dioxygen moiety. Besides the eta(1)-end-on iron(III)-superoxo complex, eta(2) side-on type iron(III)-superoxo complex is also optimized. This species is more stable than the eta(1)-end-on type iron(III)-superoxo complex, suggesting that this is considered as a stable isomer in the early stage of the catalytic cycle. PMID- 19284796 TI - Expedient preparation of all isomers of 2-aminocyclobutanecarboxylic acid in enantiomerically pure form. AB - A short, convenient, gram scale protocol has been established to allow facile access to all four stereoisomers of 2-aminocyclobutanecarboxylic acid, each in enantiomerically pure form (ee >99%). Starting from the readily available cis racemate, the procedure combines efficient alpha-phenylethylamine derivative resolution and controlled cis-to-trans epimerization procedures, and proceeds with invariably high yields. PMID- 19284797 TI - Understanding the mechanism of drug resistance due to a codon deletion in protoporphyrinogen oxidase through computational modeling. AB - Protoporphyrinogen oxidase (PPO; EC 1.3.3.4) is the last common enzyme for the enzymatic transformation of protoporphyrinogen-IX to protoporphyrin-IX, which is the key common intermediate leading to heme and chlorophyll. Hence, PPO has been identified as one of the most importance action targets for the treatment of some important diseases including cancer and variegated porphyria (VP). In the agricultural field, PPO inhibitors have been used as herbicides for many years. Recently, a unique drug resistance was found to be associated with a nonactive site residue (Gly210) deletion rather than substitution in A. tuberculatus PPO. In the present study, extensive computational simulations, including homology modeling, molecular dynamics (MD) simulations, and molecular mechanics-Poisson Boltzmann surface area (MM-PBSA) calculations, have been carried out to uncover the detailed molecular mechanism of drug resistance associated with Gly210 deletion. Although Gly210 in the wild-type A. tuberculatus PPO has no direct interaction with the inhibitors, all the computational models and energetic results indicated that Gly210 deletion has great effects on the hydrogen-bonding network and the conformational change of the binding pocket. An interchain hydrogen bond between Gly210 with Ser424, playing an important role in stabilizing the local conformation of the wild-type enzyme, disappeared after Gly210 deletion. As a result, the mutant-type PPO has a lower affinity than the wild-type enzyme, which accounts for the molecular mechanism of drug resistance. The structural and mechanistic insights obtained from the present study provide a new starting point for future rational design of novel PPO inhibitors to overcome drug resistance associated with Gly210 deletion. PMID- 19284798 TI - Large-scale synthesis and microwave absorption enhancement of actinomorphic tubular ZnO/CoFe2O4 nanocomposites. AB - Actinomorphic tubular ZnO/CoFe(2)O(4) nanocomposites were fabricated in large scale via a simple solution method at low temperature. The phase structures, morphologies, particle size, shell thickness, chemical compositions of the composites have been characterized by X-ray diffraction (XRD), field emission scanning electron microscope (FESEM), energy dispersive X-ray spectroscopy (EDS), and transmission electron microscopy (TEM). The as-synthesized nanocomposites were uniformly dispersed into the phenolic resin then the mixture was pasted on metal plate with the area of 200 mm x 200 mm as the microwave absorption test plate. The test of microwave absorption was carried out by the radar-absorbing materials (RAM) reflectivity far field radar cross-section (RCS) method. The range of microwave absorption is from 2 to 18 Hz and the best microwave absorption reach to 28.2 dB at 8.5 Hz. The results indicate that the composites are of excellence with respect to microwave absorption. PMID- 19284799 TI - Involvement of ethylene in the accumulation of esculeoside A during fruit ripening of tomato (Solanum lycopersicum). AB - The composition of glycoalkaloids in tomato fruit changes with ripening. However, it has not been clarified whether the accumulation of glycoalkaloids is controlled by the ripening-inducing phytohormone, ethylene. Here, we report the effect of ethylene on the accumulation of tomato fruit glycoalkaloids. We investigated the effect of exogenously applied ethylene. In response to ethylene treatment, the content of alpha-tomatine decreased, whereas the content of esculeoside A increased. Next, we analyzed the fruits of ripening mutants, rin, nor, and Nr. In fruits of these mutant lines, the level of accumulation of esculeoside A decreased, whereas alpha-tomatine accumulated to higher levels than in wild-type fruit. These results demonstrated that the esculeoside A accumulation was associated with production and perception of ethylene. Additionally, the accumulation profiles of the intermediate metabolites of esculeoside A biosynthesis in ripening mutant fruits suggest that a glycosylation step in the putative pathway from alpha-tomatine to esculeoside A depends on ethylene. PMID- 19284800 TI - Characterization of a rice bran oil structured lipid. AB - Rice bran oil (RBO) was enzymatically modified in a continuous packed bed bioreactor to incorporate caprylic acid with Lipozyme RM IM as biocatalyst. The reaction product was purified by short-path distillation. Rice bran oil structured lipid (RBOSL) contained 32.1 mol % caprylic acid. Positional analysis revealed 0.7 mol % caprylic acid at the sn-2 position and 47.8 mol % caprylic acid at the sn-1,3 positions. Composition of free fatty acids and smoke point of RBO and RBOSL were not significantly different. Saponification value, iodine value, and viscosity of RBO were significantly different from those of RBOSL. The color of RBOSL was darker, more yellow and less green than RBO. Volatile compounds in RBO and RBOSL were determined by GC-MS. Melting onset temperatures of RBO and RBOSL were not significantly different, while melting end point temperatures and melting enthalpies were significantly different. This characterization study results will help determine potential food applications of RBOSL. PMID- 19284801 TI - Two cases of primary yolk sac tumor of the liver in childhood: case reports and literature review. AB - Primary yolk sac tumor of the liver is an extremely rare neoplasm, with fewer than 20 cases reported. We evaluated 2 pediatric cases (21-month-old and 23-month old female patients), who presented with liver masses and markedly raised serum alpha-fetoprotein (AFP). One patient received a partial hepatectomy and another patient underwent a liver biopsy, both of which showed characteristic features of yolk sac tumor, with tumor cells staining strongly positive for AFP. There was no evidence of an extrahepatic primary source. Both of our patients have been healthy, without evidence of recurrent tumor, and their AFP level remains in the normal range after their chemotherapeutic treatments. PMID- 19284802 TI - Morphology and small-subunit ribosomal DNA sequence of Henneguya adiposa (Myxosporea) from Ictalurus punctatus (Siluriformes). AB - The original description of Henneguya adiposa, a myxozoan parasitizing channel catfish Ictalurus punctatus, is supplemented with new data on spore morphology, including photomicrographs and line drawings, as well as 18S small-subunit (SSU) ribosomal DNA (rDNA) sequence. Elongate, translucent, linear plasmodia were situated between the connective tissue bands of the adipose fin. The myxospore possessed an elongate, lanceolate spore body, flattened parallel to the suture line: 17.1 +/- 1.6 microm (range 14.7-20.5) in length, 4.1 +/- 0.3 microm (range 3.4-4.6) wide in valvular view, and 3.7 +/- 0.3 microm (range 3.2-4.0) thick in sutural view. Polar capsules were pyriform and unequal in length, measuring 7.2 +/- 0.6 microm (range 5.8-8.3) long and 1.3 +/- 0.2 microm (range 0.9-1.9) wide, with at least 8 turns in the polar filament coil. The caudal appendages were 38.0 +/- 6.2 microm (range 23.2-48.8) and split posteriorly. The total length of the spore was 55.6 +/- 6.5 microm (range 40.7-65.8). Phylogenetic analysis of 18S SSU rDNA sequence data placed this isolate within the Myxobolidae, indicating a close relationship to a group of Henneguya species parasitizing channel catfish in the southeast United States. PMID- 19284803 TI - Comparison of passive fecal flotation run by veterinary students to zinc-sulfate centrifugation flotation run in a diagnostic parasitology laboratory. AB - The sensitivity of fecal examination methods can be influenced by both technician error and methodology. In this analysis, we compared the results of 335 passive fecal flotation examinations performed on the feces of stray dogs by 3rd-yr veterinary students at the University of Pennsylvania, School of Veterinary Medicine, to the results obtained through zinc-sulfate centrifugation performed by the diagnostic parasitology laboratory on the same fecal samples. The students' passive flotation results agreed with the laboratory zinc-sulfate centrifugation for only 62.4% of samples. Students were able to diagnose 75.0% of Ancylostoma caninum cases, 71.4% of Toxocara canis cases, 54.2% of Trichuris vulpis cases, 26.7% of Cystoisospora spp. (C. ohioensis-like and C. canis) cases, and 14.7% of Giardia lamblia cases. There were also 70 instances where students reported the presence of parasites in the sample that were not diagnosed by zinc sulfate centrifugation. Based on the overall study findings, passive fecal flotation examinations run in private practice could be missing up to 50.5% of infected dogs, due to either technician error or inherent limitations to the passive fecal flotation technique. PMID- 19284805 TI - A blueprint for pharmacy benefit managers to increase value. AB - Pharmacy benefit managers (PBMs) have a unique opportunity to promote health and generate value in the healthcare system. Today, PBMs are largely evaluated on their ability to control costs rather than improve health. Pharmacy benefit managers should be evaluated along 3 dimensions in which they can increase value: (1) use of cost-effective medications, (2) timely initiation of appropriate medication therapy, and (3) adherence to that therapy. Value creation requires the development of integrated data systems, stronger partnerships with patients and physicians, and improved measurement and reporting of results. Incentives for PBMs to promote value should drive innovation and improve health outcomes. PMID- 19284806 TI - Cost-effectiveness of long-term oxygen therapy for chronic obstructive disease. AB - OBJECTIVES: To assess the cost-effectiveness of long-term oxygen therapy to facilitate proper resource allocation. STUDY DESIGN: Markov process. METHODS: A Markov model was developed to estimate the incremental cost-effectiveness ratios (ICERs) for continuous and nocturnal oxygen therapies. The maximum time horizon was set to 5 years. Efficacy variables were obtained from pertinent clinical studies. Cost variables were based on the current Medicare reimbursement rate and on appropriate sources. Multiple 1-way and probabilistic sensitivity analyses were performed to examine the robustness of base-case results. RESULTS: The ICER for continuous oxygen therapy ($16,124 per quality-adjusted life-year [QALY]) was within bounds considered to be cost-effective, while that of nocturnal oxygen therapy was not ($306,356/QALY). The estimated ICER for continuous oxygen therapy was robust (95% confidence interval, $13,153-$24,658/QALY) and was more favorable than the ICERs for commonly used medical and surgical therapies for chronic obstructive pulmonary disease. The ICER for nocturnal oxygen therapy was sensitive to variation in the mortality rate; it could be as low as $18,267/QALY gained. At the other end, nocturnal oxygen therapy could be less effective than no oxygen therapy, despite additional costs. CONCLUSIONS: There is substantial room for improvement in the current Medicare policies regarding long-term oxygen therapy. Medicare coverage can be improved by prescribing long-term oxygen therapy to patients who will receive substantial benefit and by providing adequate support for services and maintenance. PMID- 19284807 TI - "No-shows": who fails to follow up with initial behavioral health treatment? AB - OBJECTIVE: We sought to determine what demographic and clinical factors are associated with receipt of initial mental health treatment. STUDY DESIGN AND METHODS: A total of 1177 patients completed structured clinical interviews (Michigan Screening for Treatment and Research Triage) when they called to authorize mental health benefits. Measures included age, sex, alcohol use, drug use, anxiety, depression, medical history, behavioral health treatment history, psychosocial stressors, functioning, and suicidality. Multivariate analyses determined the association between these variables and a behavioral health claim within 90 days of the interview. RESULTS: Among those completing interviews, 85% attended initial mental health treatment. Factors significantly associated with increased odds of treatment initiation were good self-rated health (odds ratio [OR] = 1.70; 95% confidence interval [CI] = 1.15, 2.50), support of family or friends (OR = 1.71; 95% CI = 1.11, 2.65), previous outpatient mental health visits (OR = 1.56; 95% CI = 1.11, 2.19), and recent alcohol use (OR = 1.41; 95% CI = 1.00, 1.97). Factors associated with decreased odds of treatment initiation were recent period of total disability (OR = 0.62; 95% CI = 0.45, 0.87), any previous suicide attempt (OR = 0.56; 95% CI = 0.36, 0.87), 6 or more physician visits for medical reasons this year (OR = 0.64; 95% CI = 0.44, 0.92), and legal problems (OR = 0.31; 95% CI = 0.16, 0.61). In multivariate analyses, family support, history of medical visits, and recent alcohol use were no longer significant predictors. CONCLUSIONS: Most individuals in this insured population who completed an initial telephone assessment had an initial behavioral health claim. However, patients with greater health or social service needs were at higher risk for not obtaining treatment, suggesting the need for greater outreach and attention by providers and insurers. PMID- 19284808 TI - Impact of 2 employer-sponsored population health management programs on medical care cost and utilization. AB - BACKGROUND: Integrated health management programs combining disease prevention and disease management services, although popular with employers, have been insufficiently researched with respect to their effect on costs. OBJECTIVE: To estimate the overall impact of a population health management program and its components on cost and utilization. STUDY DESIGN, SETTING, AND PARTICIPANTS: Observational study of 2 employer-sponsored health management programs involving more than 200,000 health plan members. METHODS: We used claims data for the first program year and the 2 preceding years to calculate cost and utilization metrics, and program activity data to determine program uptake. Using an intent-to-treat approach and regression-based risk adjustment, we estimated whether the program was associated with changes in cost and utilization. Data on program fees were unavailable. RESULTS: Overall, the program was associated with a nonsignificant cost increase of $13.75 per member per month (PMPM). The wellness component alone was associated with a significant increase of $20.14 PMPM. Case and disease management were associated with a significant decrease in hospital admissions of 4 and 1 per 1000 patient-years, respectively. CONCLUSIONS: Our results suggest that the programs did not reduce medical cost in their first year, despite a beneficial effect on hospital admissions. If we had been able to include program fees, it is likely that the overall cost would have increased significantly. Although this study had important limitations, the results suggest that a belief that these programs will save money may be too optimistic and better evaluation is needed. PMID- 19284809 TI - The effects of antihypertensive step-therapy protocols on pharmaceutical and medical utilization and expenditures. AB - OBJECTIVE: To examine the effects of antihypertensive step therapy on prescription drug utilization and spending, and other medical care utilization and spending. STUDY DESIGN: Pre/post design. METHODS: Employers who had implemented step therapy were compared with employers who had not implemented step therapy. Data were drawn from the 2003 through 2006 MarketScan Research Databases. The study sample included employees and dependents who used antihypertensives (11,851 patients whose employer implemented a step-therapy protocol and 30,882 patients in the comparison group without step therapy). Multivariate generalized estimating equation models were used to estimate the immediate and time-varying effects of step therapy on medical and prescription drug spending and utilization, while controlling for important covariates and adjusting for clustering by patient. RESULTS: Results showed an initial 7.9% reduction in antihypertensive medication days supplied and an initial 3.1% reduction in medication costs among antihypertensive users in the step-therapy plans. However, these percentages grew in each subsequent quarter. Antihypertensive users in step-therapy programs also experienced an increase in inpatient admissions and emergency room visits. After an initial decline in spending, the step-therapy group incurred $99 more per user in quarterly expenditures than the comparison group. CONCLUSIONS: The intended effect of step therapy is to substitute cheaper and equivalently effective medications for more expensive medications. As this study demonstrates, step therapy may create barriers to receiving any medication, resulting in higher medical utilization and costs. Further research is needed to understand why these unintended consequences occur and how they might be avoided. PMID- 19284810 TI - Provider organization performance assessment utilizing diabetes physician recognition program. AB - OBJECTIVE: To examine the level of care in a large provider organization with respect to the Diabetes Physician Recognition Program (DPRP) standards of care and describe treatment patterns, diabetes-related complications, and achievement of clinical goals among patients stratified by glycemic control levels. STUDY DESIGN: Observational, retrospective, chart review study assessing care among patients with diabetes. METHODS: Diabetic patients aged > or =5 years who were prescribed insulin or oral hypoglycemics/antihyperglycemics for at least 12 months by 1 of 14 DPRP-participating physicians from the provider organization were eligible. A sample of patient medical charts was collected. Descriptive statistics were generated to assess demographic and clinical variables, with subanalyses for patients in the 3 glycosylated hemoglobin (A1C) cohorts. Data were used to describe the demographics, disease prevalence, comorbidities, clinical outcomes, and treatment patterns of the study population. Results were assessed according to national treatment guidelines. RESULTS: Almost all DPRP recognition measures were met and/or exceeded. More than 90% of patients received appropriate assessments. The majority achieved the A1C, blood pressure, and low density lipoprotein control levels recommended by national treatment guidelines. Patients with multiple comorbidities had worse levels of control, with only 14.3% of patients achieving all 3 treatment goals. Nearly 30% of patients had diabetes related complications, most commonly kidney disease. CONCLUSIONS: Differences in control and treatment patterns exist in patients with varying levels of glycemic control. Opportunities exist to improve diabetes care through goal attainment. Further research is needed to determine whether specific measures of care correlate with levels of glycemic control. PMID- 19284811 TI - Reporting hospitals' antibiotic timing in pneumonia: adverse consequences for patients? AB - OBJECTIVE: To determine whether publicly reporting hospital scores on antibiotic timing in pneumonia (percentage of patients with pneumonia receiving antibiotics within 4 hours) has led to unintended adverse consequences for patients. STUDY DESIGN: Retrospective analyses of 13,042 emergency department (ED) visits by adult patients with respiratory symptoms in the National Hospital Ambulatory Medical Care Survey, 2001-2005. METHODS: Rates of pneumonia diagnosis, antibiotic use, and waiting times to see a physician were compared before and after public reporting, using a nationally representative hospital sample. These outcomes also were compared between hospitals with different antibiotic timing scores. RESULTS: There were no differences in rates of pneumonia diagnosis (10% vs 11% of all ED visits, P = .72) or antibiotic administration (34% vs 35%, P = .21) before and after antibiotic timing score reporting. Mean waiting times to be seen by a physician increased similarly for patients with and without respiratory symptoms (11-minute vs 6-minute increase, respectively; P = .29). After adjustment for confounders, hospitals with higher 2005 antibiotic timing scores had shorter mean waiting times for all patients, but there were no significant score-related trends for rates of pneumonia diagnosis or antibiotic use. CONCLUSION: Despite concerns, public reporting of hospital antibiotic timing scores has not led to increased pneumonia diagnosis, antibiotic use, or a change in patient prioritization. PMID- 19284877 TI - Microdeletion syndromes disclose replication timing alterations of genes unrelated to the missing DNA. AB - BACKGROUND: The temporal order of allelic replication is interrelated to the epigenomic profile. A significant epigenetic marker is the asynchronous replication of monoallelically-expressed genes versus the synchronous replication of biallelically-expressed genes. The present study sought to determine whether a microdeletion in the genome affects epigenetic profiles of genes unrelated to the missing segment. In order to test this hypothesis, we checked the replication patterns of two genes - SNRPN, a normally monoallelically expressed gene (assigned to 15q11.13), and the RB1, an archetypic biallelically expressed gene (assigned to 13.q14) in the genomes of patients carrying the 22q11.2 deletion (DiGeorge/Velocardiofacial syndrome) and those carrying the 7q11.23 deletion (Williams syndrome). RESULTS: The allelic replication timing was determined by fluorescence in situ hybridization (FISH) technology performed on peripheral blood cells. As expected, in the cells of normal subjects the frequency of cells showing asynchronous replication for SNRPN was significantly (P < 10-12) higher than the corresponding value for RB1. In contrast, cells of the deletion-carrying patients exhibited a reversal in this replication pattern: there was a significantly lower frequency of cells engaging in asynchronous replication for SNRPN than for RB1 (P < 10-4 and P < 10-3 for DiGeorge/Velocardiofacial and Williams syndromes, respectively). Accordingly, the significantly lower frequency of cells showing asynchronous replication for SNRPN than for RB1 is a new epigenetic marker distinguishing these deletion syndrome genotypes from normal ones. CONCLUSION: In cell samples of each deletion-carrying individual, an aberrant, reversed pattern of replication is delineated, namely, where a monoallelic gene replicates more synchronously than a biallelic gene. This inverted pattern, which appears to be non-deletion-specific, clearly distinguishes cells of deletion-carriers from normal ones. As such, it offers a potential epigenetic marker for suspecting a hidden microdeletion that is too small to be detected by conventional karyotyping methods. PMID- 19284879 TI - Sexual violence in the protracted conflict of DRC programming for rape survivors in South Kivu. AB - BACKGROUND: Despite international acknowledgement of the linkages between sexual violence and conflict, reliable data on its prevalence, the circumstances, characteristics of perpetrators, and physical or mental health impacts is rare. Among the conflicts that have been associated with widespread sexual violence has been the one in the Democratic Republic of the Congo (DRC). METHODS: From 2003 till to date Malteser International has run a medico-social support programme for rape survivors in South Kivu province, DRC. In the context of this programme, a host of data was collected. We present these data and discuss the findings within the frame of available literature. RESULTS: Malteser International registered 20,517 female rape survivors in the three year period 2005-2007. Women of all ages have been targeted by sexual violence and only few of those - and many of them only after several years - sought medical care and psychological help. Sexual violence in the DRC frequently led to social, especially familial, exclusion. Members of military and paramilitary groups were identified as the main perpetrators of sexual violence. CONCLUSION: We have documented that in the DRC conflict sexual violence has been - and continues to be - highly prevalent in a wide area in the East of the country. Humanitarian programming in this field is challenging due to the multiple needs of rape survivors. The easily accessible, integrated medical and psycho-social care that the programme offered apparently responded to the needs of many rape survivors in this area. PMID- 19284878 TI - Nitric oxide activates ATP-sensitive potassium channels in mammalian sensory neurons: action by direct S-nitrosylation. AB - BACKGROUND: ATP-sensitive potassium (KATP) channels in neurons regulate excitability, neurotransmitter release and mediate protection from cell-death. Furthermore, activation of KATP channels is suppressed in DRG neurons after painful-like nerve injury. NO-dependent mechanisms modulate both KATP channels and participate in the pathophysiology and pharmacology of neuropathic pain. Therefore, we investigated NO modulation of KATP channels in control and axotomized DRG neurons. RESULTS: Cell-attached and cell-free recordings of KATP currents in large DRG neurons from control rats (sham surgery, SS) revealed activation of KATP channels by NO exogenously released by the NO donor SNAP, through decreased sensitivity to [ATP]i. This NO-induced KATP channel activation was not altered in ganglia from animals that demonstrated sustained hyperalgesia type response to nociceptive stimulation following spinal nerve ligation. However, baseline opening of KATP channels and their activation induced by metabolic inhibition was suppressed by axotomy. Failure to block the NO-mediated amplification of KATP currents with specific inhibitors of sGC and PKG indicated that the classical sGC/cGMP/PKG signaling pathway was not involved in the activation by SNAP. NO-induced activation of KATP channels remained intact in cell-free patches, was reversed by DTT, a thiol-reducing agent, and prevented by NEM, a thiol-alkylating agent. Other findings indicated that the mechanisms by which NO activates KATP channels involve direct S-nitrosylation of cysteine residues in the SUR1 subunit. Specifically, current through recombinant wild-type SUR1/Kir6.2 channels expressed in COS7 cells was activated by NO, but channels formed only from truncated isoform Kir6.2 subunits without SUR1 subunits were insensitive to NO. Further, mutagenesis of SUR1 indicated that NO-induced KATP channel activation involves interaction of NO with residues in the NBD1 of the SUR1 subunit. CONCLUSION: NO activates KATP channels in large DRG neurons via direct S-nitrosylation of cysteine residues in the SUR1 subunit. The capacity of NO to activate KATP channels via this mechanism remains intact even after spinal nerve ligation, thus providing opportunities for selective pharmacological enhancement of KATP current even after decrease of this current by painful-like nerve injury. PMID- 19284880 TI - Identification of candidate genes for human pituitary development by EST analysis. AB - BACKGROUND: The pituitary is a critical neuroendocrine gland that is comprised of five hormone-secreting cell types, which develops in tandem during the embryonic stage. Some essential genes have been identified in the early stage of adenohypophysial development, such as PITX1, FGF8, BMP4 and SF-1. However, it is likely that a large number of signaling molecules and transcription factors essential for determination and terminal differentiation of specific cell types remain unidentified. High-throughput methods such as microarray analysis may facilitate the measurement of gene transcriptional levels, while Expressed sequence tag (EST) sequencing, an efficient method for gene discovery and expression level analysis, may no-redundantly help to understand gene expression patterns during development. RESULTS: A total of 9,271 ESTs were generated from both fetal and adult pituitaries, and assigned into 961 gene/EST clusters in fetal and 2,747 in adult pituitary by homology analysis. The transcription maps derived from these data indicated that developmentally relevant genes, such as Sox4, ST13 and ZNF185, were dominant in the cDNA library of fetal pituitary, while hormones and hormone-associated genes, such as GH1, GH2, POMC, LHbeta, CHGA and CHGB, were dominant in adult pituitary. Furthermore, by using RT-PCR and in situ hybridization, Sox4 was found to be one of the main transcription factors expressed in fetal pituitary for the first time. It was expressed at least at E12.5, but decreased after E17.5. In addition, 40 novel ESTs were identified specifically in this tissue. CONCLUSION: The significant changes in gene expression in both tissues suggest a distinct and dynamic switch between embryonic and adult pituitaries. All these data along with Sox4 should be confirmed to further understand the community of multiple signaling pathways that act as a cooperative network that regulates maturation of the pituitary. It was also suggested that EST sequencing is an efficient means of gene discovery. PMID- 19284881 TI - A clinical evaluation committee assessment of recombinant human tissue factor pathway inhibitor (tifacogin) in patients with severe community-acquired pneumonia. AB - INTRODUCTION: The purpose of this analysis was to determine the potential efficacy of recombinant human tissue factor pathway inhibitor (tifacogin) in a subpopulation of patients with community-acquired pneumonia (CAP) from a phase III study of severe sepsis. METHODS: A retrospective review of patients with suspected pneumonia was conducted by an independent clinical evaluation committee (CEC) blinded to treatment assignment. The CEC reanalyzed data from patients enrolled in an international multicenter clinical trial of sepsis who had a diagnosis of pneumonia as the probable source of sepsis. The primary efficacy measure was all-cause 28-day mortality. RESULTS: Of 847 patients identified on case report forms with a clinical diagnosis of pneumonia, 780 (92%) were confirmed by the CEC to have pneumonia. Of confirmed pneumonia cases, 496 (63.6%) met the definition for CAP. In the CEC CAP population, the mortality rates of the tifacogin and placebo groups were 70/251 (27.9%) and 80/245 (32.7%), respectively. The strongest signals were seen in patients with CAP not receiving concomitant heparin, having microbiologically confirmed infection, or having the combination of documented infection and no heparin. The reduction in mortality in this narrowly defined subgroup when treated with tifacogin compared with placebo was statistically significant (17/58 [29.3%] with tifacogin and 28/54 [51.9%] with placebo; unadjusted P value of less than 0.02). CONCLUSIONS: Tifacogin administration did not significantly reduce mortality in any severe CAP patient. Exploratory analyses showed an improved survival in patients who did not receive concomitant heparin with microbiologically confirmed infections. These data support the rationale of an ongoing phase III study exploring the potential benefit of tifacogin in severe CAP. TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT00084071. PMID- 19284882 TI - Loop-mediated isothermal amplification applied to filarial parasites detection in the mosquito vectors: Dirofilaria immitis as a study model. AB - BACKGROUND: Despite recent advances in our understanding of the basic biology behind transmission of zoonotic infectious diseases harbored by arthropod vectors these diseases remain threatening public health concerns. For effective control of vector and treatment, precise sampling indicating the prevalence of such diseases is essential. With an aim to develop a quick and simple method to survey zoonotic pathogen-transmitting vectors, LAMP (loop-mediated isothermal amplification) was applied to the detection of filarial parasites using a filarial parasite-transmitting experimental model that included one of the mosquito vectors, Aedes aegypti, and the canine heartworm, Dirofilaria immitis. RESULTS: LAMP reactions amplifying the cytochrome oxidase subunit I gene demonstrated high sensitivity when a single purified D. immitis microfilaria was detected. Importantly, the robustness of the LAMP reaction was revealed upon identification of an infected mosquito carrying just a single parasite, a level easily overlooked using conventional microscopic analysis. Furthermore, successful detection of D. immitis in wild-caught mosquitoes demonstrated its applicability to field surveys. CONCLUSION: Due to its simplicity, sensitivity, and reliability, LAMP is suggested as an appropriate diagnostic method for routine diagnosis of mosquito vectors carrying filarial parasites. This method can be applied to the survey of not only canine filariasis but also lymphatic filariasis, another major public health problem. Therefore, this method offers great promise as a useful diagnostic method for filarial parasite detection in endemic filariasis regions. PMID- 19284883 TI - Activation of an AMP-activated protein kinase is involved in post-diapause development of Artemia franciscana encysted embryos. AB - BACKGROUND: Cysts of Artemia can remain in a dormant state for long periods with a very low metabolic rate, and only resume their development with the approach of favorable conditions. The post-diapause development is a very complicated process involving a variety of metabolic and biochemical events. However, the intrinsic mechanisms that regulate this process are unclear. RESULTS: Herein we report the specific activation of an AMP-activated protein kinase (AMPK) in the post diapause developmental process of Artemia. Using a phospho-AMPKalpha antibody, AMPK was shown to be phosphorylated in the post-diapause developmental process. Results of kinase assay analysis showed that this phosphorylation is essential for AMPK activation. Using whole-mount immunohistochemistry, phosphorylated AMPK was shown to be predominantly located in the ectoderm of the early developed embryos in a ring shape; however, the location and shape of the activation region changed as development proceeded. Additionally, Western blotting analysis on different portions of the cyst extracts showed that phosphorylated AMPKalpha localized to the nuclei and this location was not affected by intracellular pH. Confocal microscopy analysis of immunofluorescent stained cyst nuclei further showed that AMPKalpha localized to the nuclei when activated. Moreover, cellular AMP, ADP, and ATP levels in developing cysts were determined by HPLC, and the results showed that the activation of Artemia AMPK may not be associated with cellular AMP:ATP ratios, suggesting other pathways for regulation of Artemia AMPK activity. CONCLUSION: Together, we report evidence demonstrating the activation of AMPK in Artemia developing cysts and present an argument for its role in the development-related gene expression and energy control in certain cells during post-diapause development of Artemia. PMID- 19284884 TI - The C-terminal sequence of the large hepatitis delta antigen is variable but retains the ability to bind clathrin. AB - BACKGROUND: Hepatitis delta virus (HDV) is a defected RNA virus and requires its encoded large antigen (LDAg) to interact with helper viral proteins (HBsAgs) during assembly. Recently, a study demonstrated a direct binding of the LDAg C terminus from genotype I HDV to the clathrin heavy chain (CHC), which suggests that this interaction might facilitate HDV assembly. If LDAg binding to clathrin is essential to HDV life cycle, a clathrin box sequence at the C-terminus of LDAg should be conserved across all HDV. However, the C-terminal sequence of LDAg is variable among 43 HDV isolates. RESULTS: Based on the presence and location of clathrin box at the C-terminus of LDAg from 43 isolates of HDV, we classified them into three groups. Group 1 (13 isolates) and 2 (26 isolates) contain a clathrin box located at amino acids 199-203 and 206-210, respectively, as found in genotype I and genotype II. Group 3 (4 isolates) contains no clathrin box as found in genotype III. CHC binding by three different LDAg (genotype I to III) was then tested by in vivo and in vitro experiments. Transfection of plasmids which encode fusion proteins of EGFP and full-length of LDAg from three genotypes into HuH-7 cells, a human hepatoma cell line, was performed. GFP-pull down assays showed that a full-length of CHC was co-precipitated by EGFP-LDI, -LDII and LDIII but not by EGFP. Further in vitro studies showed a full-length or fragment (amino acids 1 to 107) of CHC can be pull-down by 13-amino-acid peptides of LDAg from three genotypes of HDV. CONCLUSION: Both in vivo and in vitro studies showed that CHC can bind to various sequences of LDAg from the three major genotypes of HDV. We therefore suggest that the clathrin-LDAg interaction is essential to the HDV life-cycle and that sequences binding to clathrin are evolutionarily selected, but nonetheless show the diversity across different HDV genotypes. PMID- 19284885 TI - Quantitative analysis of CT-perfusion parameters in the evaluation of brain gliomas and metastases. AB - BACKGROUND: The paper reports a quantitative analysis of the perfusion maps of 22 patients, affected by gliomas or by metastasis, with the aim of characterizing the malignant tissue with respect to the normal tissue. The gold standard was obtained by histological exam or nuclear medicine techniques. The perfusion scan provided 11 parametric maps, including Cerebral Blood Volume (CBV), Cerebral Blood Flow (CBF), Average Perfusion (Pmean) and Permeability-surface area product (PS). METHODS: The perfusion scans were performed after the injection of 40 ml of non-ionic contrast agent, at an injection rate of 8 ml/s, and a 40 s cine scan with 1 s interval was acquired. An expert radiologist outlined the region of interest (ROI) on the unenhanced CT scan, by using a home-made routine. The mean values with their standard deviations inside the outlined ROIs and the contralateral ROIs were calculated on each map. Statistical analyses were used to investigate significant differences between diseased and normal regions. Receiving Operating Characteristic (ROC) curves were also generated. RESULTS: Tumors are characterized by higher values of all the perfusion parameters, but after the statistical analysis, only the PS, PatRsq (Patlak Rsquare) and Tpeak (Time to Peak) resulted significant. ROC curves, confirmed both PatRsq and PS as equally reliable metrics for discriminating between malignant and normal tissues, with areas under curves (AUCs) of 0.82 and 0.81, respectively. CONCLUSION: CT perfusion is a useful and non invasive technique for evaluating brain neoplasms. Malignant and normal tissues can be accurately differentiated using perfusion map, with the aim of performing tumor diagnosis and grading, and follow-up analysis. PMID- 19284886 TI - Induction of lipid oxidation by polyunsaturated fatty acids of marine origin in small intestine of mice fed a high-fat diet. AB - BACKGROUND: Dietary polyunsaturated fatty acids (PUFA), in particular the long chain marine fatty acids docosahexaenoic (DHA) and eicosapentaenoic (EPA), are linked to many health benefits in humans and in animal models. Little is known of the molecular response to DHA and EPA of the small intestine, and the potential contribution of this organ to the beneficial effects of these fatty acids. Here, we assessed gene expression changes induced by DHA and EPA in the wildtype C57BL/6J murine small intestine using whole genome microarrays and functionally characterized the most prominent biological process. RESULTS: The main biological process affected based on gene expression analysis was lipid metabolism. Fatty acid uptake, peroxisomal and mitochondrial beta-oxidation, and omega-oxidation of fatty acids were all increased. Quantitative real time PCR, and -- in a second animal experiment -- intestinal fatty acid oxidation measurements confirmed significant gene expression differences and showed in a dose-dependent manner significant changes at biological functional level. Furthermore, no major changes in the expression of lipid metabolism genes were observed in the colon. CONCLUSION: We show that marine n-3 fatty acids regulate small intestinal gene expression and increase fatty acid oxidation. Since this organ contributes significantly to whole organism energy use, this effect on the small intestine may well contribute to the beneficial physiological effects of marine PUFAs under conditions that will normally lead to development of obesity, insulin resistance and diabetes. PMID- 19284887 TI - Human parainfluenza virus type 2 (HPIV2) induced host ADAM8 expression in human salivary adenocarcinoma cell line (HSY) during cell fusion. AB - BACKGROUND: The aim of the study was to investigate expression of ADAMs (A Disintegrin and A Metalloproteinase) of host cell origin during cell-cell fusion induced by human parainfluenza virus type 2 (HPIV2). RESULTS: Induction of host cell ADAM9 was observed in GMK cells, but the applicability of this model was restricted by lack of cross-reactivity of the anti-human ADAM8 antibodies with the corresponding green monkey antigens. HSG cells were not susceptible to HPIV2 virus infection. In contrast, in human parotid gland HSY cells, a natural host cell for paramyxoviruses, HPIV2 induced ADAM8 expression. ADAM8 staining increased dramatically over time from 7.9 +/- 3% at zero hours to 99.2 +/- 0.8% at 72 hours (p = 0.0001). Without HPIV2 the corresponding percentages were only 7.7% and 8.8%. Moreover, ADAM8 positive cells formed bi- (16.2%) and multinuclear cells (3.5%) on day one and the corresponding percentages on day three were 15.6% for binuclear and 57.2% for multinuclear cells. CONCLUSION: ADAM8, well recognized for participation in cell-to-cell fusion especially in osteoclast formation, is up-regulated upon formation of multinuclear giant cells after HPIV2 induction in HSY cells. The virus-HSY cell system provides a novel experimental model for study of the molecular mechanism of cell fusion events. PMID- 19284888 TI - Myxobolus cerebralis (Myxozoa), the causative agent of whirling disease, reduces fecundity and feeding activity of Tubifex tubifex (Oligochaeta). AB - Myxobolus cerebralis is the causative agent of whirling disease that has significant economical and ecological impacts on trout populations. Although intensive studies have been conducted to understand its effects on and interactions with its fish host, only limited information is available about how and to what extent M. cerebralis affects its oligochaete host, Tubifex tubifex. We investigated the effects of M. cerebralis on survival, growth, reproduction, and feeding activity of T. tubifex. Mature, immature and juvenile worms were exposed to myxospores and their infection prevalence, mortality, sexual development, reproduction and spore production were compared with unexposed worms. The parasite affected neither survival nor growth but inhibited clitellar development and reduced cocoon production by over 80%. Numbers of actinospores released from mature worms were nearly 9-fold higher than that of immature worms. When non-clitellated infected worms were kept at 30 degrees C for 4 days, spore release ceased and they re-developed a clitellum. These results suggest parasite induced castration. Comparative monitoring of defecation rate revealed that M. cerebralis reduced feeding activity of T. tubifex by approximately 40%. Low energy intake and impaired energetic allocation may be the underlying mechanism behind reduced fecundity of infected T. tubifex. PMID- 19284889 TI - Electron channeling: a problem for x-ray microanalysis in materials science. AB - Electron channeling effects can create measurable signal intensity variations in all product signals that result from the scattering of the electron beam within a crystalline specimen. Of particular interest to the X-ray microanalyst are any variations that occur within the characteristic X-ray signal that are not directly related to a specimen composition variation. Many studies have documented the effect of crystallographic orientation on the local X-ray yield; however, the vast majority of these studies were carried out on thin foil specimens examined in transmission. Only a few studies have addressed these effects in bulk specimen materials, and these analyses were generally carried out at common scanning electron microscope microanalysis overvoltages (>1.5). At these overvoltage levels, the anomalous transmission effect is weak. As a result, the effect of electron channeling on the characteristic X-ray signal intensity has traditionally been overlooked in the field of quantitative electron probe microanalysis. The present work will demonstrate that electron channeling can produce X-ray variations of up to 26%, between intensity maxima and minima, in low overvoltage X-ray microanalyses of bulk specimens. Intensity variations of this magnitude will significantly impact the accuracy of qualitative and quantitative X-ray microanalyses at low overvoltage on engineering structural materials. PMID- 19284890 TI - Detector solid angle formulas for use in X-ray energy dispersive spectrometry. AB - With the advent of silicon drift X-ray detectors, a range of new geometries has become possible in electron optical columns. Because of their compact size, these detectors can potentially achieve high geometrical collection efficiencies; however, using traditional approximations detector solid angle calculations rapidly break down and at times can yield nonphysical values. In this article we present generalized formulas that can be used to calculate the variation in detection solid angle for contemporary Si(Li) as well as new silicon drift configurations. PMID- 19284891 TI - X-ray microanalysis combined with monte carlo simulation for the analysis of layered thin films: the case of carbon contamination. AB - A previously developed Monte Carlo code has been extended to the X-ray microanalysis in a (scanning) transmission electron microscope of plan sections, consisting of bilayers and triple layers. To test the validity of this method for quantification purposes, a commercially available NiOx (x 1) thin film, deposited on a carbon layer, has been chosen. The composition and thickness of the NiO film and the thickness of the C support layer are obtained by fitting to the three X ray intensity ratios I(NiK)/I(OK), I(NiK)/I(CK), and I(OK)/I(CK). Moreover, it has been investigated to what extent the resulting film composition is affected by the presence of a contaminating carbon film at the sample surface. To this end, the sample has been analyzed both in the (recommended) "grid downward" geometry and in the upside/down ("grid upward") situation. It is found that a carbon contaminating film of few tens of nanometers must be assumed in both cases, in addition to the C support film. Consequently, assuming the proper C/NiOx/C stack in the simulations, the Monte Carlo method yields the correct oxygen concentration and thickness of the NiOx film. PMID- 19284892 TI - Chemical shift of electron energy-loss near-edge structure on the nitrogen K-edge and titanium L3-edge at TiN/Ti interface. AB - We investigated the chemical shift of the electron energy-loss near-edge structure (ELNES) for the nitrogen K-edge and titanium L3-edge measured from the interface region between a titanium nitride layer and a titanium layer. Both the titanium nitride and titanium layers were prepared by a sputtering method. Elemental analysis for nitride and titanium in the vicinity of the interface region was performed using a standard technique in electron energy-loss spectroscopy. It was demonstrated that both the ELNES of nitrogen K-edge and titanium L3-edge presented the chemical shift, more or less, depending on the composition of TiNx. The experimental findings were interpreted using a first principles band structure calculation. The chemical shifts of nitrogen K-edge and titanium L3-edge can be used as fingerprinting for readily distinguishing the composition of TiNx. PMID- 19284893 TI - A transmission electron microscopy study of Fe-Co alloy nanoparticles in silica aerogel matrix using HREM, EDX, and EELS. AB - Magnetic nanocomposite materials consisting of 5.5 wt% Fe-Co alloy nanoparticles in a silica aerogel matrix, with compositions Fe(x)Co(1-x) of x = 0.50 and 0.67, have been synthesized by the sol-gel method. The high-resolution transmission electron microscopy images show nanoparticles consisting of single crystal grains of body-centered cubic Fe-Co alloy, with typical crystal grain diameters of approximately 4 and 7 nm for Fe(0.5)Co(0.5) and Fe(0.67)Co(0.33) samples, respectively. The energy dispersive X-ray (EDX) spectra summed over areas of the samples gave compositions Fe(x)C(o1-x) with x = 0.48 +/- 0.06 and 0.68 +/- 0.05. The EDX spectra obtained with the 1.5 nm probe positioned at the centers of approximately 20 nanoparticles gave slightly lower concentrations of Fe, with means of x = 0.43 +/- 0.01 and x = 0.64 +/- 0.02, respectively. The Fe(0.5)Co(0.50) sample was studied using electron energy loss spectroscopy (EELS), and EELS spectra summed over whole nanoparticles gave x = 0.47 +/- 0.06. The EELS spectra from analysis profiles of nanoparticles show a distribution of Fe and Co that is homogeneous, i.e., x = 0.5, within a precision of at best +/ 0.05 in x and +/-0.4 nm in position. The present microscopy results have not shown the presence of a thin layer of iron oxide, but this might be at the limit of detectability of the methods. PMID- 19284894 TI - Secondary electron emission contrast of quantum wells in GaAs p-i-n junctions. AB - The secondary electron (SE) signal over a cleaved surface of GaAs p-i-n solar cells containing stacks of quantum wells (QWs) is analyzed by high-resolution scanning electron microscopy. The InGaAs QWs appear darker than the GaAsP barriers, which is attributed to the differences in electron affinity. This method is shown to be a powerful tool for profiling the conduction band minimum across junctions and interfaces with nanometer resolution. The intrinsic region is shown to be pinned to the Fermi level. Additional SE contrast mechanisms are discussed in relation to the dopant regions themselves as well as the AlGaAs window at the p-region. A novel method of in situ observation of the SE profile changes resulting from reverse biasing these structures shows that the built-in potential may be deduced. The obtained value of 0.7 eV is lower than the conventional bulk value due to surface effects. PMID- 19284895 TI - Toward reproducible three-dimensional microstructure analysis of granular materials and complex suspensions. AB - Focused ion beam nanotomography (FIB-nt) is a novel method for high resolution three-dimensional (3D) imaging. In this investigation we assess the methodological parameters related to image acquisition and data processing that are critical for obtaining reproducible microstructural results from granular materials and from complex suspensions. For this purpose three case studies are performed: (1) The precision of FIB-nt is evaluated by analyzing a reference sample with nanospheres. Due to the implementation of an automated correction procedure, drift phenomena can be removed largely from the FIB data. However, at high magnifications remaining drift components can induce problems for 3D-shape reconstructions. (2) Correct object recognition from densely packed microstructures requires specific algorithms for splitting of agglomerated particles. To establish quantitative criteria for the correct degree of splitting, a parametric study with dry portland cement is performed. It is shown that splitting with a k-value of 0.6 leads to accurate results. (3) Finally, the reproducibility of the entire cryo-FIB analysis is investigated for high pressure frozen cement suspensions. Reproducible analyses can be obtained if the magnification is adapted to the particle size. At low magnifications the small particles and their surface area are underestimated. At high magnifications representativity is questioned because local inhomogeneities can become dominant. PMID- 19284896 TI - Contrast mechanisms and image formation in helium ion microscopy. AB - The helium ion microscope is a unique imaging instrument. Based on an atomic level imaging system using the principle of field ion microscopy, the helium ion source has been shown to be incredibly stable and reliable, itself a remarkable engineering feat. Here we show that the image contrast is fundamentally different to other microscopes such as the scanning electron microscope (SEM), although showing many operational similarities due to the physical ion interaction mechanisms with the sample. Secondary electron images show enhanced surface contrast due the small surface interaction volume as well as elemental contrast differences, such as for nanowires imaged on a substrate. We present images of nanowires and nanoparticles for comparison with SEM imaging. Applications of Rutherford backscattered ion imaging as a unique and novel imaging mechanism are described. The advantages of the contrast mechanisms offered by this instrument for imaging nanomaterials are clearly apparent due to the high resolution and surface sensitivity afforded in the images. Future developments of the helium ion microscope should yield yet further improvements in imaging and provide a platform for continued advances in microscope science and nanoscale research. PMID- 19284897 TI - Ultrastructure of the epithelial cells associated with tooth biomineralization in the chiton Acanthopleura hirtosa. AB - The cusp epithelium is a specialized branch of the superior epithelium that surrounds the developing teeth of chitons and is responsible for delivering the elements required for the formation of biominerals within the major lateral teeth. These biominerals are deposited within specific regions of the tooth in sequence, making it possible to conduct a row by row examination of cell development in the cusp epithelium as the teeth progress from the unmineralized to the mineralized state. Cusp epithelium from the chiton Acanthopleura hirtosa was prepared using conventional chemical and microwave assisted tissue processing, for observation by light microscopy, conventional transmission electron microscopy (TEM) and energy filtered TEM. The onset of iron mineralization within the teeth, initiated at row 13, is associated with a number of dramatic changes in the ultrastructure of the apical cusp cell epithelium. Specifically, the presence of ferritin containing siderosomes, the position and number of mitochondria, and the structure of the cell microvilli are each linked to aspects of the mineralization process. These changes in tissue development are discussed in context with their influence over the physiological conditions within both the cells and extracellular compartment of the tooth at the onset of iron mineralization. PMID- 19284898 TI - Scanning electron microscopy and synchrotron radiation x-ray tomographic microscopy of 330 million year old charcoalified seed fern fertile organs. AB - Abundant charcoalified seed fern (pteridosperm) pollen organs and ovules have been recovered from Late Visean (Mississippian 330 Ma) limestones from Kingswood, Fife, Scotland. To overcome limitations of data collection from these tiny, sometimes unique, fossils, we have combined low vacuum scanning electron microscopy on uncoated specimens with backscatter detector and synchrotron radiation X-ray tomographic microscopy utilizing the Materials Science and TOMCAT beamlines at the Swiss Light Source of the Paul Scherrer Institut. In combination these techniques improve upon traditional cellulose acetate peel sectioning because they enable study of external morphology and internal anatomy in multiple planes of section on a single specimen that is retained intact. The pollen organ Melissiotheca shows a basal parenchymatous cushion bearing more than 100 sporangia on the distal face. Digital sections show the occurrence of pollen in some sporangia. The described ovule is new and has eight integumentary lobes that are covered in spirally arranged glandular hairs. Virtual longitudinal sections reveal the lobes are free above the pollen chamber. Results are applied in taxonomy and will subsequently contribute to our understanding of the former diversity and evolution of ovules, seeds, and pollen organs in the seed ferns, the first seed-bearing plants to conquer the land. PMID- 19284899 TI - Practice, politics, and possibilities. PMID- 19284900 TI - [Single-stranded electronic medical record--paradigmatic shift that has failed to appear]. PMID- 19284901 TI - [Assessment of a single-stranded electronic patient record]. AB - INTRODUCTION: Patient care and treatment is usually documented in a double stranded patient record, i.e. a record with separate sections for different health professionals, which reduces the possibility of getting a comprehensive view of the patient's case. Therefore, the Pediatric Department chose to implement a single-stranded medical record. MATERIAL AND METHODS: Development and evaluation of the new record was based on standards formulated during interdisciplinary audits. Eighteen months after its implementation, the record was evaluated by interdisciplinary audits and questionnaires. All health professionals employed at the department were included. Data from the questionnaires were analyzed descriptively and summaries of the audits were analyzed using an anthropological method. RESULTS: A total of 149 staff members (96%) responded to the questionnaire and eight records were evaluated by audits at which a total of 63 interdisciplinary clinicians participated. The evaluation concluded that overlapping documentation was reduced considerably, 97% of the staff members reported that their documentation was being read by their colleagues and 84% reported that the patient record gave them a good general view of the patients' case apart from complicated patient cases and long lasting admissions. CONCLUSION: The single-stranded interdisciplinary patient record reduces documentation overlap. The record facilitates overview and continuity in short-term patient cases. The participating staff became acquainted with and now uses the information documented by other health professionals. PMID- 19284902 TI - [Insufficient blood pressure control following coronary revascularization]. AB - INTRODUCTION: After coronary revascularization, anti-angina treatment is often withdrawn or reduced while the ejection fraction tends to improve. Both contribute to increase blood pressure (BP). The purpose of the present study was to evaluate BP control in patients after revascularization procedures. MATERIAL AND METHODS: In 190 patients (48 females) who had passed a coronary revascularization procedure at least 12 months earlier, the blood pressure was measured both at the ambulatory and at the clinic; blood samples were drawn for determination of lipids and plasma glucose; body mass index (BMI) was determined; the clinical history was taken with special emphasis on medication and smoking habits. RESULTS: The mean BP was 134.2 +/- 17.6/81.1 +/- 9.7 mmHg at the clinic and the daytime ambulatory value was 132.8 +/- 13.8/78.3 +/- 8.7 mmHg. Poor BP control was found in 42.6% of the patients according to clinic measurements and in 51.6% according to ambulatory measurements. Among the patients with known diabetes, 64.3% were poorly controlled according to clinic measurements and 75.0% according to ambulatory measurements. Medicines with BP lowering effects were taken by 79.0% of the patients. BMI was above 25 in 71.6% of the patients, whereas 58.9% of the patients had total cholesterol >or= 4.5 mmol/l. Twenty-eight percent were still smokers a year after revascularization. CONCLUSION: The BP control in patients after revascularization is unsatisfactory as approximately half of the study participants had a BP above the recommended ranges. PMID- 19284903 TI - [Patient information leaflets seen through the eyes of patients in a general practice]. AB - INTRODUCTION: The objective was to investigate patients' perception of patient information leaflets, including their readability, comprehensibility and their impact on adherence. MATERIAL AND METHODS: A 14-item questionnaire was prepared and answered by unselected patients in a general practice in Copenhagen. RESULTS: A total of 111 patients were included. Eighty-eight patients (79%) always or often read the patient information leaflets. Among these, 77% attached importance to adverse effects. A mere 13% attached importance to all leaflet information. In all, 21% never or rarely read patient information leaflets, but relied on the doctor or the pharmacist. 62% of the interviewed had no problems reading or understanding the patient information leaflets. Among the 38% with reading problems, 57% had difficulties due to text-size, and 33% found the written language difficult to understand. Thirty-five patients (32%) stated that they had stopped taking medication due to the information about adverse effects. CONCLUSION: The patient information leaflet is an important source of drug information as most patients read the leaflet and nearly a third of the patients stated that information about the adverse effects had made them stop taking their medicine. Future studies should bring into focus the reason for medication adherence, how written information can be made easier to read, comprehensive and correct without contributing to anxiety and non-adherence. PMID- 19284904 TI - [Pregnancy after Mustard operation for transposition of the great arteries]. AB - INTRODUCTION: Previously, the Mustard operation was the preferred surgical technique in patients with transposition of the great arteries. After this procedure the anatomical right ventricle remains the systemic ventricle, which entails long-term complications, especially heart failure. The Mustard-operated patients are now adults, and pregnancy has become an important issue. We assessed pregnancy and delivery data in a complete cohort of female patients who had previously undergone a Mustard procedure at Aarhus Hospital in the 1971-1991 period. The women giving birth were compared with those who did not. MATERIAL AND METHODS: Information on diagnosis, procedure, functional status, pregnancy, delivery and congenital heart disease in the off-spring were found in registers (The National Register of Health, The National Birth Register) and in medical records. Questionnaires were mailed to the 16 surviving women over 15 years of age. RESULTS: Four women gave birth to nine children. Two women temporally had heart symptoms during pregnancy and delivery, but this caused no objective changes. The women who had children were older than those who did not. None of the children had congenital heart disease nor were they small for their gestational age. CONCLUSION: Pregnancy and delivery are well-tolerated in the vast majority of Mustard-operated women with no deterioration in functional class during or after pregnancy and delivery. PMID- 19284905 TI - [Psychological debriefing]. AB - Diagnostic criteria for stress reactions and adjustment disorders are summarized. Existing psychological treatments are outlined with a focus on evidence-based methods, and recommendations for treatment are described. The conclusion is that there is no evidence for the effect of psychological debriefing or other professional treatment in the immediate aftermath of traumatic events. On the other hand, cognitive behavioural therapy, EMDR and similar methods with focus on exposure can be recommended, both as a preventive strategy and for patients with post-traumatic stress disorder. PMID- 19284906 TI - [Argatroban treatment of heparin-induced immune-mediated thrombocytopenia]. AB - Heparin-induced immune-mediated thrombocytopenia (HIT) is a life-threatening complication of heparin treatment. When HIT is clinically suspected, heparin treatment should immediately be replaced with an alternative, fast-acting, anticoagulant agent, and blood tests should be carried out to verify or to exclude the diagnosis. Argatroban has recently been approved for HIT treatment in Denmark. Recommendations for dosing of argatroban in HIT patients with and without comorbities are presented. PMID- 19284907 TI - [Heparin-induced thrombocytopenia]. AB - Heparin treatment can cause an immune-mediated thrombocytopenia: HIT. HIT antibodies can be detected by various methods, but laboratory analyses are not specific or sensitive and may delay the diagnostic process. It is therefore important to initiate alternative treatment based on the clinical findings, and a clinical score system for evaluating the risk of HIT has been suggested. When HIT is likely, treatment consists of immediate replacement of heparin with alternative anticoagulation treatment and refrainment from warfarin therapy and platelet infusion. PMID- 19284908 TI - [Surgical treatment of diabetes mellitus type 2]. AB - The resolution of diabetes after gastric bypass is already observed during the first days after the operation, while the control of diabetes after gastric banding occurs after weight loss. Other mechanisms than weight loss must therefore explain the improved glucose tolerance. Focus has been on appetite regulating gut hormone, particularly peptide YY and glucagon-like peptide-1. Nevertheless, other unknown hormones or mechanisms may be involved in the dramatic changes in the glucose metabolism. Development of new drugs and treatments based on the endocrine changes observed after bariatric surgery with the aim of curing type 2-diabetes are warranted. PMID- 19284909 TI - [Loss of sight and hearing and cognitive dysfunction - Susac's syndrome?]. AB - Cognitive dysfunction, unexplained deafness or retinal branch artery occlusions in otherwise healthy persons are challenging conditions, in particular when occurring in young adults. Susac's syndrome is characterized by the clinical triad of encephalopathy, hearing loss and retinal branch arterial occlusions affecting mainly young women. Appropriate diagnosis and management requires multidisciplinary management of the disease. PMID- 19284910 TI - [Complications of unnecessary arterial puncture]. AB - This case describes the complications of an arterial puncture in an 83-year-old female with a femoral neck fracture. Two arterial punctures were performed in the perioperative period. Ten days after surgery the patient was re-admitted with a pseudo aneurysm in the radial artery above the wrist. The artery was ligated and no signs of infection were found. Arterial puncture is an important source of information, but it is essential to use the correct aseptic technique and to apply compression to the artery afterwards. The indication should always be considered, as a venous blood sample can often provide the information needed. PMID- 19284911 TI - [Intrauterine pregnancy following balloon thermal endometrial ablation (Cavaterm)]. AB - A case of intrauterine pregnancy occurring after successful balloon thermal endometrial ablation is described. Although rare, pregnancy after endometrial ablation is possible, and use of a supplemental contraceptive method should be planned. In case of pregnancy after endometrial ablation, the woman should be informed of the high risk of pregnancy complications, and termination of the pregnancy should be discussed. PMID- 19284912 TI - [Uterine rupture in a primigravid woman at 31st gestational week]. AB - Spontaneous uterine rupture is a very rare obstetric complication. This case report describes spontaneous uterine rupture in a healthy 28-year-old primigravid woman at the 31st gestational week. The woman was hospitalized due to contractions, which eased after rest. Later she had acute abdominal pain followed by an abnormal cardiotocography. An acute caesarean section was performed and a longitudinal posterior uterine wall rupture found. She had no known risk factors explaining the rupture. PMID- 19284913 TI - [Picture of the month: lymphogranuloma venereum]. PMID- 19284914 TI - [Transfusion in Denmark]. PMID- 19284915 TI - [Transfusion practice in total hip arthroplasty in Danish departments of orthopaedic surgery]. AB - INTRODUCTION: We examined the use of blood transfusion in patients undergoing total hip arthroplasty (THA) at 21 orthopaedic departments in Denmark. MATERIAL AND METHODS: Patients with primary THA (n = 21,773) between 1999 and 2006 were identified from the Danish Hip Arthroplasty Registry. Data on the use of blood transfusion was collected from the Danish Transfusion Database (DTDB). The outcome was defined as red blood cell transfusion (yes/no) within eight days of surgery. We estimated the relative risk for red blood cell transfusion (RR) and a 95% confidence interval (CI) adjusting for possible confounding factors, including patient- and surgery-related factors. The risk of blood transfusion for each department was compared with the overall risk of blood transfusion for all other departments. RESULTS: Overall, red blood cell transfusion was given to 8,198 of 21,960 patients (37%) (range: 16-64%). The adjusted RRs varied from 0.5 (95% CI, 0.4-0.7) to 1.8 (95% CI, 1.4-2.5) using all departments as reference. The coefficient of variation was 33% based on crude relative risk estimates and dropped to 31% after adjustment for patient- and surgery-related factors. CONCLUSIONS: Substantial differences in the use of red blood cell transfusion among THA patients were found when comparing a sample of Danish orthopaedic departments. The differences in the use of blood transfusions could apparently not be explained by a range of patient- and surgery-related factors, which suggests that the variation did not reflect differences in the patients' need for blood transfusion, but rather true differences in transfusion practice. PMID- 19284916 TI - [Hand washing prevents diarrhea. A survey of a Cochrane review]. PMID- 19284917 TI - [Fusobacterium necrophorum: from tonsillitis to Lemierre's syndrome]. AB - Fusobacterium necrophorum plays a role in non-GAS-tonsillitis in adolescents and probably also in small children with recurrent otitis media. Anaerobic culture on selective anaerobic media is recommended to detect F. necrophorum and treat such patients. Denmark sees at least 20 annual cases of Lemierre's syndrome with a mortality reaching 9%. Early suspicion of Lemierre's syndrome in adolescents with non-GAS-tonsillitis, who develop septicaemia, pulmonary symptoms and unilateral swelling on the neck, is mandatory to lower morbidity and mortality. PMID- 19284918 TI - [Can anaerobic culture of throat swabs prevent Lemierre's syndrome?]. AB - Six cases of Lemierre's syndrome were reported from 2004 to 2007 at Viborg Hospital, corresponding to 33 cases per year in Denmark. All six patients were healthy younger persons presenting with a suspected bacterial tonsillitis which had been found strep A antigen negative. Fusobacterium necrophorum was found in throat swabs by anaerobic culture on selective media and/or by real-time PCR. We recommend that all patients 10 to 40 years of age with strep A antigentest negative bacterial tonsillitis have throat swabs anaerobically cultured on selective media. We believe early identification and therapy may prevent progression to Lemierre's syndrome. PMID- 19284919 TI - [Late sequelae following electrical accidents]. AB - A literature review of non-acute pain, neurological and psycho-neurological impairments in relation to electrical accidents is performed. Pain in extremities is frequent. Neurological and psycho-neurological symptoms can arise days or months after an electrical accident. The causal mechanisms are not known. The best known non-organic effect is post-traumatic stress disorder. The existing knowledge of long-term consequences of electrical accidents is generally based on cases and patient summaries. There is serious lack of longitudinal studies describing delayed or progressive non-acute symptoms. PMID- 19284920 TI - [Diagnostic classification in Denmark with emphasis on general practice]. AB - Diagnostic classification may be used as a structure for the naming of health problems and to create an overview in electronic patient records. Classification also facilitates access to health information and decision support, quality improvement and research. The International Classification Primary Care is now included in the WHO's Family of International Classifications and was introduced into Danish general practice in 1998. This system will be updated to ICPC-2-DK in 2009. In secondary health care, the ICD-10 is currently in use but a new system (SNOMED-CT) is underway. PMID- 19284921 TI - [Vestibular nerve section in the treatment of morbus Meniere]. AB - INTRODUCTION: Vestibular nerve section in the treatment of Mb. Meniere was introduced in Denmark in 1980. This treatment is centralised at Gentofte Hospital and we have previously published the results from 1980-1996. We here present the updated results from 2000-2007 with a total of 18 patients. MATERIAL AND METHODS: Systematic review of case journals and questionnaires regarding postoperative satisfaction, vertigo control and influence on daily activities. RESULTS: All patients achieved total vertigo spell control. In total 15 patients (83%) indicated satisfaction with the operation. Sixteen patients (89%) reported that vertigo had no, mild, or moderate influence on their daily activities. The difference between pre- and postoperative functional level was highly significant. Expected consequences of unilateral ablation in the form of imbalanced gait or lingering sensation of dizziness occurred to varying degrees in 14 patients (78%). COMPLICATIONS: Two patients developed postoperative liquorrhoea and one patient partial facial palsy. CONCLUSION: The results demonstrate that vestibular nerve section causes vertigo spell control and improved daily functional levels in patients with severe and otherwise untreatable Mb. Meniere. These results are in accordance with international publications. New prospects in vestibular rehabilitation promise significant reductions of the postoperative imbalance. Continued centralisation of the surgical intervention is recommended, as is providing patients with information of the positive results of this treatment. PMID- 19284922 TI - [Mini clinical evaluation exercise as evaluation tool of communicative and cooperative skills in the outpatient clinic]. AB - INTRODUCTION: In the revised Danish medical specialist training increased focus has been placed on competences which are hard to evaluate such as communication skills. Mini-CEX seems promising as an evaluation tool. Our aim was to test: 1) whether mini-CEX was useable in the evaluation of communicative and cooperative skills and 2) whether mini-CEX would provide reproducible data. MATERIAL AND METHODS: Twenty-one residents were evaluated by mini-CEX by trained observers. Seventeen residents had at least two observations within a short period of time and these data were used to estimate the mini-CEX reproducibility. In addition to the residents, the nurses who assisted them in the outpatient clinic answered a questionnaire regarding the mini-CEX satisfaction. RESULTS: Observations had a median duration of 20 minutes (10-60 minutes) and the overall median duration of feedback was 15 minutes (5-60 minutes). Time used for feedback was halved from the first to the following feedback sessions. No significant clinical differences were observed between the scorings performed by the residents themselves and the observers, or the nurses of the outpatient clinic and the observers. In general, the residents were satisfied with the mini-CEX evaluations. CONCLUSION: The mini CEX is a promising tool for the evaluation of communicative and cooperative skills. PMID- 19284923 TI - [Developments in the population's use of the internet for health-related purposes]. AB - INTRODUCTION: We wish to study the development in the Danish population's use of the internet for health-related purposes. What has changed from 2005 to 2007? Who is the typical user and what exactly is the internet used for? MATERIAL AND METHODS: As part of a larger EU-funded survey, 1,000 randomly selected Danes between 15-80 years of age completed a questionnaire in the fall 2005. The same questions were put to 1.021 Danes in the spring of 2007 to study any trends. RESULTS: A total of 72% of the Danes use the internet when searching for information on health. This constitutes an increase of 20% from 2005. Personal contact with the general practitioner (GP) is still the most important source of health and illness information, but more than 50% also consider the internet to be an important or very important source of health information. Women are still more active than men when searching for health information. A total of 20% of the Danes have consulted their own GP online, which is a 161% increase. Also, it has become more important for the Danes when choosing a GP that he or she offers eServices. CONCLUSION: The Danes' use of the internet for health information purposes has increased. And when the Danes seek health and illness information, it is primarily used to decide if the GP should be consulted. The GP remains the primary source of information and the Danes consult with their GP more and more frequently via the internet. PMID- 19284924 TI - [Dermoid cyst presenting with atypical upper extremity neurological symptoms]. AB - Dermoid cysts, also known as teratodermoids, are benign tumours thought to originate from multipotent embryonic cells which constitute approx. 10% of all mediastinal tumours. This is a rare presentation of a mediastinal cystic teratoma presenting with radiating pain and disturbed sensitivity in the right upper extremity. These tumours are generally benign and surgery is the treatment of choice. PMID- 19284925 TI - [Septic shock caused by Gardnerella vaginalis and Peptostreptococcus species after Cesarean section]. AB - A 41-year old multisectio patient got a life-threatening postoperative infection with Gardnerella vaginalis and Peptostreptococcus spp. Perioperative treatment with cefuroxime and metronidazol is recommended. PMID- 19284926 TI - [Can acute renal insufficiency increase the effect of warfarin?]. AB - The use of warfarin is complicated by its narrow therapeutic index. The risk of severe complications in the form of haemorrhage is significantly increased by an elevated degree of anticoagulation. We present a case in which a patient in stable warfarin treatment was admitted with acute renal failure and extremely high international normalized ratio (INR). No single mechanism can fully explain this. We find that it was probably caused by accumulation of active metabolites, an inhibitory factor in uraemic blood, which reduces hepatic metabolism and a genotype with an unstable metabolism of warfarin. PMID- 19284927 TI - Safety and subjective sleep effects of ramelteon administration in adults and older adults with chronic primary insomnia: a 1-year, open-label study. AB - OBJECTIVE: To evaluate the long-term safety and subjective sleep effects of ramelteon in adults with chronic insomnia. METHOD: Subjects with primary insomnia (DSM-IV-TR criteria) for >or= 3 months received ramelteon nightly for 1 year; a 3 day placebo run out followed. Subjects aged >or=65 years received open-label ramelteon 8 mg (N = 248); those aged 18 to 64 years received ramelteon 16 mg (N = 965). Subjects completed sleep diaries and returned to the clinic at week 1 and at months 1, 2, 3, 4, 6, 8, 10, and 12 for safety assessments and investigator performed Clinical Global Impressions. The study was conducted from February 2003 through September 2004. RESULTS: There were no noteworthy changes in vital signs, physical examinations, clinical chemistry, hematology, or urinalysis values and no electrocardiogram changes to suggest adverse cardiac effects. Endocrine values remained within normal range throughout treatment. Consistent statistically significant (p or= 10% of OFC patients were weight gain, increased appetite, dry mouth, somnolence, fatigue, headache, and peripheral edema. Random glucose mean change (mg/dL) was +7.92 for the OFC group, +1.62 for the fluoxetine group (p = .020), and +9.91 for the olanzapine group (p = .485). Random cholesterol mean change (mg/dL) was +12.4 for OFC, +2.3 for fluoxetine (p < .001), and +3.1 for olanzapine (p < .001); incidence of treatment-emergent increase from normal to high cholesterol (baseline < 200 mg/dL and >or= 240 subsequently) was significantly higher for the OFC group (10.2%) than for the fluoxetine group (3.1%, p = .017) but not the olanzapine group (8.0%, p = .569). Mean weight change (kg) was +4.42 for OFC, -0.15 for fluoxetine (p < .001), and +4.63 for olanzapine (p = .381), with 40.4% of OFC patients gaining >or= 7% body weight (vs. olanzapine: 42.9%, p = .515; fluoxetine: 2.3%, p < .001). CONCLUSION: Results of this analysis showed that OFC-treated patients experienced significantly improved depressive symptoms compared with olanzapine- or fluoxetine-treated patients following failure of 2 or more antidepressants within the current depressive episode. Safety results for OFC were generally consistent with those for its component monotherapies. The total cholesterol increase associated with OFC was more pronounced than with olanzapine alone. PMID- 19284929 TI - Long-term mental health resource utilization and cost of care following group psychoeducation or unstructured group support for bipolar disorders: a cost benefit analysis. AB - OBJECTIVE: To explore the short- and long-term mental health resource utilization and cost of care in a sample of 120 individuals with bipolar disorders who participated in a randomized controlled efficacy trial of group psychoeducation versus unstructured group support. METHOD: Prospective, independent monitoring of DSM-IV bipolar disorder type I or II patients aged 18 to 65 years was conducted during the intervention phase (6 months) and follow-up phase (5-year postintervention) of a randomized controlled trial reporting clinical outcomes and inpatient and outpatient mental health service utilization, with estimation of cost of treatment per patient. The study was conducted from October 1997 through October 2006. RESULTS: Compared with individuals with bipolar disorder receiving the control intervention, psychoeducated patients had twice as many planned outpatient appointments, but the estimated mean cost of emergency consultation utilization was significantly less. There were trends for psychoeducated patients to opt for self-funded psychotherapy after completing group psychoeducation and to utilize more medications. However, inpatient care accounted for 40% estimated total cost in the control group but only about 15% in the psychoeducation group. CONCLUSIONS: This study demonstrates the importance of taking a long-term overview of the cost versus benefits of adjunctive psychological therapy in bipolar disorders. If viewed only in the short-term, the psychoeducation group used more mental health care resources without clear additional health gain. However, extended follow-up demonstrated a long-term advantage for psychoeducated individuals, such that, compared to an unstructured support group intervention, group psychoeducation is less costly and more effective. PMID- 19284930 TI - Comparing the effectiveness of aripiprazole and quetiapine in schizophrenia and related psychoses: a naturalistic, retrospective chart review study. AB - BACKGROUND: Naturalistic studies offer advantages over randomized clinical trials by including patients seen in routine practice. Aripiprazole and quetiapine are the most recent second-generation antipsychotics available in the United Kingdom. We aimed to study all patients who were prescribed these medications in a defined geographic area in order to identify and compare those who had a good clinical response. METHOD: We conducted an electronic chart review of a sample of all people attending secondary mental health care in the county of Lanarkshire, Scotland, who were treated with aripiprazole or quetiapine for schizophrenia and related psychoses (ICD-10 criteria) between 2002 and 2007. To measure effectiveness, we retrospectively assigned Clinical Global Impressions (CGI) scores and examined medication discontinuation rates. RESULTS: Eighty-nine patients were started on treatment with aripiprazole and 132 patients with quetiapine over the 5-year period. Those treated with quetiapine had a higher initial illness severity (CGI-Severity of Illness scale) (p = .0003), were more likely to be starting rather than switching antipsychotics (p = .0003), were more likely to have a mood disorder (p = .03), were less likely to be treatment resistant (p = .005), and had lower rates of prescription of additional antipsychotics (p = .009). After adjusting for these variables, the proportions who improved according to CGI were 74% with aripiprazole and 67% with quetiapine. Overall medication discontinuation rates were also similar, 42% for aripiprazole and 45% for quetiapine, with early discontinuation of aripiprazole being noticeable, often due to agitation (13% of all patients treated with the drug). CONCLUSIONS: Despite their different pharmacologic properties, aripiprazole and quetiapine were similarly effective in the majority of patients. Early discontinuation of aripiprazole due to agitation was an important finding. PMID- 19284931 TI - Risk factors for violent crime in Schizophrenia: a national cohort study of 13,806 patients. AB - OBJECTIVE: To determine risk factors for and prevalence of violent crime in patients with schizophrenia, and in particular, to explore the contribution of familial risk factors. METHOD: We designed a cohort study that followed up patients with 2 or more hospitalizations for schizophrenia (ICD-8, ICD-9, and ICD 10 criteria) and investigated the risk for a violent conviction using Cox proportional hazards models. All 13,806 patients with 2 hospital discharge diagnoses of schizophrenia from January 1, 1973, through December 31, 2004, in Sweden were followed until violent conviction, emigration, death, or end of follow-up (December 31, 2004), and associations with sociodemographic, individual (substance abuse comorbidity, and previous violence), and familial (parental violent crime and parental alcohol abuse) factors were examined. RESULTS: Over an average follow-up period of 12 years, 17.1% (N = 1519) of the men and 5.6% (N = 273) of the women with 2 or more hospitalizations for schizophrenia had a violent conviction after discharge from hospital. Familial risk factors had moderate effects, increasing the risk for violent convictions by 50% to 150%. After adjustment for sociodemographic and individual risk factors, the associations between parental violent crime and risk of violent convictions remained in men (adjusted hazard ratio [HR] = 1.65, 95% CI = 1.33 to 2.04) and in women (adjusted HR = 1.83, 95% CI = 1.11 to 3.01), whereas parental alcohol abuse was no longer significantly associated with violent crime. CONCLUSION: Parental violent crime had moderate associations with violent crime in male and female offspring with at least 2 hospitalizations for schizophrenia, which were mostly stronger than the better documented sociodemographic risk factors. This suggests that familial (genetic or early environmental) risk factors have an important role in the etiology of violent offending among individuals with schizophrenia and should be considered in violence risk assessment. PMID- 19284932 TI - Sexual function in postpartum women treated for depression: results from a randomized trial of nortriptyline versus sertraline. AB - OBJECTIVE: The primary aim of this article is to describe sexual concerns in postpartum women with DSM-IV diagnoses of major depressive disorder (MDD) before and during treatment with antidepressants in an 8-week double-blind randomized trial. METHOD: Seventy women aged 19-42 years participated and were randomly assigned to either the tricyclic antidepressant nortriptyline (N = 38) or the serotonin selective reuptake inhibitor sertraline (N = 32). Women completed the Arizona Sexual Experience Scale to evaluate sexual concerns at enrollment and weekly during the trial. The outcome measure for depression, Hamilton Rating Scale for Depression, was completed in clinical interviews at the same time points. Comparisons of demographic and other characteristics of women were completed with t tests for continuous measures and with chi-squared or Fisher exact statistics for categorical measures. Mixed-effects regressions were used to test for significance of the main effects of depression symptom scores, drug assignment, weeks treated with medication, and the interactions of these variables. Data were collected from April 1997 to April 2002. RESULTS: At entry into the randomized trial, 73% (N = 51) of the women reported problems in 3 or more areas of sexual concern compared to 37% (N = 26) at week 8. There were no significant differences at study entry in women randomly assigned to nortriptyline compared to those randomly assigned to sertraline in summary scores of sexual function nor in specific sexual concerns at any time point. At week 8, women whose MDD remitted were more likely to report fewer (< 3) sexual concerns than women whose MDD did not remit (76% vs. 24%, p = .006), independent of drug assignment. CONCLUSIONS: In postpartum women, sexual concerns are primarily affected by remission of depression rather than side effects of either a tricyclic or serotonergic antidepressant. PMID- 19284933 TI - Evidence for efficacy and tolerability of vilazodone in the treatment of major depressive disorder: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: The efficacy and tolerability of vilazodone, a combined selective serotonin reuptake inhibitor and partial 5-hydroxytryptamine-1A (5-HT(1A)) receptor agonist, were evaluated in adult patients with major depressive disorder (MDD). METHOD: This was a randomized, double-blind, placebo-controlled trial conducted from February 2006 to May 2007. Patients aged 18 through 65 years with MDD (DSM-IV criteria) and a baseline 17-item Hamilton Rating Scale for Depression (HAM-D-17) score of >or= 22 were randomly assigned to vilazodone or placebo for 8 weeks. Vilazodone was titrated from 10 mg to 40 mg once a day over 2 weeks. Efficacy was assessed by mean change from baseline to week 8 on the Montgomery Asberg Depression Rating Scale (MADRS), HAM-D-17, and Hamilton Rating Scale for Anxiety. Response rates were determined at week 8 for the MADRS, HAM-D-17, and Clinical Global Impressions-Severity of Illness (CGI-S) and -Improvement (CGI-I) scales. Data were analyzed using a modified last-observation-carried-forward method in the intention-to-treat (ITT) sample. The Arizona Sexual Experience Scale (ASEX) was also measured at baseline and week 8. RESULTS: Of 410 randomly assigned patients, 198 receiving vilazodone and 199 receiving placebo were included in the ITT population. The mean changes in MADRS and HAM-D-17 total scores from baseline to week 8 were significantly (p = .001 and p = .022, respectively) greater with vilazodone than with placebo. Significant (p < .05) improvements in MADRS and HAM-D-17 scores were noted at week 1, the earliest time point measured. Response rates were significantly higher with vilazodone than with placebo on the MADRS (p = .007), HAM-D-17 (p = .011), and CGI-I (p = .001). Treatment-emergent adverse events with vilazodone included diarrhea, nausea, and somnolence; most adverse events were of mild or moderate intensity. There were no clinically significant differences for either gender in ASEX scores at end of treatment. CONCLUSIONS: Vilazodone is effective for the treatment of MDD in adults, with symptom relief starting at 1 week, and is well tolerated at a dose of 40 mg/day. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00285376. PMID- 19284934 TI - Guideline update. VTE disease: which agents, and when. PMID- 19284935 TI - Clinical inquiries. Does routine amniotomy have a role in normal labor? PMID- 19284936 TI - Clinical inquiries. Does reducing smoking in the home protect children from the effects of second-hand smoke? PMID- 19284937 TI - You can do more to slow the progression of heart failure. PMID- 19284938 TI - Singulair-induced anaphylaxis? PMID- 19284939 TI - Abnormal uterine bleeding: avoid the rush to hysterectomy. PMID- 19284940 TI - Arthroscopic surgery for knee osteoarthritis? Just say no. PMID- 19284941 TI - CDC recommendations expand vaccine indications. PMID- 19284942 TI - Warty papule and scaling around finger. PMID- 19284943 TI - Clinical inquiries. Can counseling prevent or treat postpartum depression? PMID- 19284944 TI - Clinical inquiries. What is the best way to manage phantom limb pain? PMID- 19284945 TI - Clinical inquiries. What treatment works best for tennis elbow? PMID- 19284946 TI - [Construction of effectiveness evaluation system for traditional Chinese medicine interventions in subhealth]. PMID- 19284947 TI - Correlation between cognitive functions and syndromes of traditional Chinese medicine in amnestic mild cognitive impairment. AB - OBJECTIVE: To explore the correlation between the cognitive functions and syndromes of traditional Chinese medicine (TCM) in amnestic mild cognitive impairment (aMCI), and to provide evidence for clinical syndrome differentiation treatment. METHODS: Six hundred subjects from Dongzhimen Hospital and seven communities in Beijing, aged between 40 and 85 years, accepted neuropsychological assessments, imaging and biochemical examinations, and syndrome differentiation, from whom 159 aMCI patients, 213 normal control (NC) subjects and 171 Alzheimer's dementia (AD) patients were screened out. Correlation between the cognitive functions and TCM syndromes in aMCI patients was analyzed. RESULTS: Mini-Mental State Examination (MMSE) score in aMCI patients was closely correlated with kidney essence vacuity and deficiency of blood and qi (r = -0.11, r = -0.11; P = 0.003, P = 0.015). Delayed Word Recall (DWR) score was correlated with kidney essence vacuity (r = -0.20, P = 0.020). Instant Story Recall (ISR) and Delayed Story Recall (DSR) scores were respectively correlated with turbid phlegm blocking upper orifices (r = -0.11, r = -0.27; P = 0.021, P = 0.000). Language function was correlated with kidney essence vacuity and deficiency of blood and qi (r = -0.11, r = -0.13; P = 0.042, P = 0.007). Attention/calculation was also closely correlated with kidney essence vacuity and deficiency of blood and qi (r = -0.10, r = -0.21; P = 0.039, P = 0.010). Attention score of aMCI patients was correlated with excess of heat toxin syndrome (r = -0.29, P = 0.026). CONCLUSION: The memory decline of aMCI is correlated with kidney essence vacuity and turbid phlegm blocking upper orifices. Furthermore, turbid phlegm blocking upper orifices is correlated with episodic memory decline, which is closely related to AD. The aMCI patients with phlegm have the risk to progress into AD. Although other cognitive functions of aMCI remain relatively intact, the patients' language function, attention/calculation and the whole cognitive function may be worsen as the aggravation of kidney essence vacuity, deficiency of blood and qi, phlegm and heat toxin, and may eventually lead to multiple cognitive domains impairment, even dementia. PMID- 19284948 TI - [Effects of Changjishu soft elastic capsule in treatment of diarrhea-predominant irritable bowel patients with liver-qi stagnation and spleen deficiency syndrome: a randomized double-blinded controlled trial]. AB - OBJECTIVE: To evaluate the efficacy and safety of Changjishu soft elastic capsule, a new Chinese herbal medicine for smoothing liver, invigorating spleen and regulating qi activity, on diarrhea-predominant irritable bowel syndrome (D IBS) patients with liver-qi stagnation and spleen deficiency syndrome. METHODS: A randomized, double-blinded, and positive control method was adopted. One hundred and four cases of D-IBS conforming to the inclusive criteria were randomly divided into treatment group (78 cases) and control group (26 cases). Patients in the treatment group were treated with Changjishu soft elastic capsule, four capsules once daily and three times a day. Patients in the control group were treated with glutamine compound enteric capsule, three capsules once daily and three times a day. After 21-day treatment, the efficacy of Changjishu soft elastic capsule was evaluated by the changes of single symptom such as abdominal pain or discomfort, defecation frequency, and stool characteristics. The safety of Changjishu elastic capsule was assessed by laboratory data and adverse events. RESULTS: Abdominal pain or discomfort scores at days 7, 14, 21 and the declined score at day 21 in the treatment group significantly decreased as compared with the control group (P < 0.05). Defecation frequency score at day 14 and the declined score at day 21 in the treatment group significantly decreased as compared with the control group (P < 0.05). Stool characteristics score and the declined score at day 21 in the treatment group significantly decreased as compared with those of the control group (P < 0.05). Rugitus scores and the declined scores at days 14 and 21 significantly decreased as compared with the control group (P < 0.05). The total response rates in the treatment and control groups were 71.62% and 40.91%, and the recovery rates were 20.27% and 13.64%, respectively. There existed significant differences between the two groups in therapeutic effect (P < 0.01). The therapeutic effect of Changjishu elastic capsule was better than that of glutamine compound enteric capsule. Side effects and adverse events were not found in the two groups. CONCLUSION: Changjishu soft elastic capsule is effective and safe for treatment of D-IBS patients with liver qi stagnation and spleen deficiency syndrome. PMID- 19284949 TI - [Changes of tongue and pulse parameters in 50 lung cancer patients treated with integrated traditional Chinese and Western medicine]. AB - OBJECTIVE: To observe the changes of tongue and pulse parameters in lung cancer patients after combined treatment with integrated traditional Chinese and Western medicine, and to probe into the application of the tongue and pulse parameters in evaluation of integrated traditional Chinese and Western medicine therapy on lung cancer patients. METHODS: Electropulsograms and tongue pictures of 50 lung cancer patients were examined by TP-I digital electropulsography. The tongue and pulse parameters of the patients, including the indexes of moistness and dryness, thinness and thickness, fissure, swollenness and emaciation, greasiness and likeness of curd, and power spectral ratios (PSRs) 1, 2, 3, 4 and cepstrum spectral ratios (CSRs) 1, 2, 3, 4, were observed after 1-month and 2-month treatment with Yifei Kangliu Oral Liquid (a compound traditional Chinese herbal medicine) plus chemotherapy, respectively. The above parameters of the lung cancer patients were compared before and after the treatment. RESULTS: PSR1 increased, while PSR2 decreased significantly after 1-month and 2-month treatment as compared with before treatment (P < 0.05). PSR1 decreased, while PSR2 increased after 2-month treatment as compared with after 1-month treatment (P < 0.05). PSR4 after 1-month treatment was lower than before treatment (P < 0.05). The tongue parameters including the indexes of moistness and dryness, and greasiness and likeness of curd increased significantly after 1-month and 2-month treatment as compared with before treatment (P < 0.05). The index of thinness and thickness decreased, while the index of fissure increased significantly after 1 month treatment as compared with before treatment (P < 0.05). CONCLUSION: Tongue and pulse parameters can be regarded as one of reference indexes in evaluation of integrated traditional Chinese and Western medicine therapy on lung cancer patients. PMID- 19284950 TI - [Effects of sophoricoside and genistein on biological characteristics of osteoblasts]. AB - OBJECTIVE: To observe the effects of sophoricoside and genistein on the biological characteristics of osteoblasts. METHODS: Osteoblasts were isolated from parietal bone of the newborn SD rats (less than 24 hours) and cultured in minimum essential medium (MEM). The effects of different concentrations of sophoricoside and genistein (10, 1 and 0.1 micromol/L) on the bone formation of osteoblasts in vitro were observed by cell proliferation rate. Differentiation of osteoblasts was evaluated by detecting the activity of alkaline phosphatase (ALP) using 4-nitrophenyl phosphate disodium salt (PNPP) method and by measuring the mineralized nodiform dyed by alizarin bordeaux. And the activity of osteoprotegerin (OPG) promoters of human beings was tested by using gene cloning and reporter gene. RESULTS: Sophoricoside at 1 and 0.1 micromol/L could promote the proliferation of osteoblasts (vs control group, P < 0.05). Genistein at 10 micromol/L could inhibit the cell proliferation, while genistein at 0.1 micromol/L showed promotion to cell proliferation (vs control group, P < 0.05). The activity of ALP of osteoblasts was increased by sophoricoside at concentrations from 0.1 micromol/L to 10 micromol/L, but genistein only at concentration of 0.1 micromol/L could make it. The total areas of mineralization of the osteoblasts in 10, 1 and 0.1 micromol/L sophoricoside groups increased by 73%, 138.6% and 114.3% as compared with the control group, respectively, but those in different concentrations of genistein groups decreased. The activity of LacZ was increased by sophoricoside at concentrations from 0.1 micromol/L to 10 micromol/L, as well as by genistein at concentration of 0.1 micromol/L. CONCLUSION: Sophoricoside can stimulate the proliferation, differentiation and mineralization of osteoblasts. And it shows no inhibition to bone formation of osteoblasts compared with genistein. Sophoricoside can also increase the activity of osteoprotegerin promoter and is more effective than genistein. PMID- 19284951 TI - [Effects of cinobufacini injection on contents of serum thyroid-stimulating hormone and adrenaline in rats]. AB - OBJECTIVE: To observe the effects of cinobufacini injection on serum levels of thyroid-stimulating hormone (TSH) and adrenaline (ADR) in rats, and to speculate the property (cold or heat) of the drug. METHODS: Thirty-two male Wistar rats were randomly divided into normal control group, cinobufacini injection group, cold syndrome group and heat syndrome group. The rats in the four groups were administered with normal saline, cinobufacini injection, and compound of traditional Chinese drugs with heat or cold property. The weights, and the serum TSH and ADR levels of the rats were measured before and after the interventions. RESULTS: The weights of the rats in heat syndrome group markedly decreased after 14-day administration when compared with the normal control group (P < 0.05). There were no significant differences among the normal control group, heat syndrome group and the cinobufacini injection group. Although no statistical difference was noted, the serum TSH levels of the cold syndrome group and cinobufacini injection group were slightly lower than that in the normal control group after 14-day administration, and the heat syndrome group showed a slightly higher serum TSH level. After administration, the serum ADR levels of the cold syndrome group and cinobufacini injection group were significantly lower than that in the normal control group (P < 0.05), and the serum ADR level in the heat syndrome group was higher than that in the normal control group (P < 0.05). CONCLUSION: Cinobufacini injection seemed to be able to suppress the thyrotrophin releasing hormone-TSH-thyroxine system and sympathetic nervous system, matching up with the characteristics of the Chinese herbs of cold property. And the study provided initial experiment evidence for the conclusion that the property of cinobufacini might be cold. PMID- 19284952 TI - [Effects of tetramethylpyrazine on angiotensin II -induced proliferation and type I collagen synthesis of rat cardiac fibroblasts]. AB - OBJECTIVE: To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type I collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin II (Ang II), and to explore the mechanism of TMP in treating myocardial fibrosis. METHODS: CFBs were isolated from neonatal rats, and the fourth-passage CFBs were used in the entire test and were stimulated by 0.1 micromol/L Ang II in vivo. The CFB proliferation was measured by methyl thiazolyl tetrazolium (MTT) assay. Type I collagen in the cell culture supernatant was measured by enzyme-linked immunosorbent assay. The expression of mRNA of type I collagen was semi-quantitatively measured by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: (1) In MTT assay, the optical density of CFBs cultured with 0.1 micromol/L Ang II was higher than that of the blank control cultured with 2% fetal bovine serum-Dulbecco's modified Eagle's medium (FBS DMEM). The difference was statistically significant (P < 0.05). Both optical densities of CFBs cultured with 0.1 micromol/L Ang II plus 800 microg/mL TMP and 0.1 micromol/L Ang II plus 600 microg/mL TMP were lower than that of CFBs cultured with 0.1 micromol/L Ang II, but only the difference between 0.1 micromol/L AngII plus 800 microg/mL TMP group and 0.1 micromol/L Ang II group was significant (P < 0.05). (2) The content of type I collagen secreted by CFBs cultured with 0.1 micromol/L Ang II was higher than that with 2% FBS-DMEM (P < 0.01). The content of type I collagen secreted by CFBs cultured with 0.1 micromol/L Ang II plus 800 microg/mL TMP was lower than that with 0.1 micromol/L Ang II (P < 0.05). (3) The level of type I collagen mRNA in 0.1 micromol/L Ang II group was higher than that in blank control group, and lower than that in 0.1 micromol/L Ang II plus 800 microg/mL TMP group. Both the differences between 0.1 micromol/L Ang II group and the blank control group and between 0.1 micromol/L Ang II group and 0.1 micromol/L Ang II plus 800 microg/mL TMP group were statistically significant (P < 0.05). CONCLUSION: TMP can not only inhibit the proliferation of CFBs, but also decrease the secretion and the mRNA expression level of collagen I in cultured CFBs of rat which are increased by Ang II. PMID- 19284953 TI - [Protective effects of salidroside on oxidative damage in fatigue mice]. AB - OBJECTIVE: To study the protective effects of salidroside on oxidative damage in fatigue mice. METHODS: Thirty-two male Kunming mice were randomly divided into four groups based on body weight: normal control group, salidroside group, training group and salidroside plus training group. The mice in the normal control group and the training group were given distilled water and mice in the salidroside group and the salidroside plus training group were given 180 mg/ (kg * d) salidroside for 15 days. At 30 min after the last administration, the mice in the training group and the salidroside plus training group were forced to swim for 120 min. Finally, all the mice were killed. The activities of lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase-myocardial band isoenzyme (CK-MB) in plasma were determined by an auto-biochemistry analyzer. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malonaldehyde (MDA) in liver tissue were also detected. The changes of ultrastructures of the skeletal muscle and cardiac muscle were observed under an electron microscope. RESULTS: Compared with no swimming, long-time swimming could significantly increase the activities of LDH, CK and CK-MB in plasma (P < 0.05, P < 0.01), while salidroside could significantly decrease the activities of CK and CK-MB in plasma induced by long-time swimming (P < 0.05, P < 0.01). There existed interactions in LDH, CK and CK-MB activities between salidroside and long-time swimming (P < 0.05). Compared with no swimming, long-time swimming could significantly decrease the SOD and GSH-Px activities and increase the MDA content in liver tissue (P < 0.01). Salidroside could significantly increase the GSH-Px and SOD activities and decrease the MDA content in liver tissue (P < 0.05, P < 0.01). However, there were no interactions in GSH-Px activity and MDA content between salidroside and long-time swimming (P < 0.05). After long-time swimming, more ultrastructural lesions were found in the cardiac muscle and skeletal muscle in the training group than in the salidroside plus training group. CONCLUSION: Salidroside may play a role in protecting the mice from oxidative damage caused by long-time endurance training. PMID- 19284954 TI - Protective effect of spleen-yin-nourishing recipe on amyloid beta-peptide-induced damage of primarily cultured rat hippocampal neurons and its mechanism. AB - OBJECTIVE: To observe the relationship among amyloid beta-peptide (Abeta)-induced neurotoxicity, serum-inducible kinase (SNK)-spine-associated Rap guanosine triphosphatase activating protein (SPAR) pathway and N-methyl-D-aspartate receptor (NMDAR), and to explore the mechanism of the protective effect of spleen yin nourishing recipe (Zibu Piyin Recipe, ZBPYR) in hippocampal neurons against Abeta-induced neurotoxicity. METHODS: The Abeta(1-40) powder was dissolved in 1 x PBS and incubated at 37 degrees centigrade, and then aggregated fibrillar Abeta(1 40) was obtained 72 h later. We used rat primary hippocampal neurons as cell model. ZBPYR-containing serum was gained by the method of serum pharmacology. ZBPYR-containing serum was added to the culture 1 h before Abeta(1-40) (5 micromol/L) exposure. Cells were harvested 2 h after Abeta(1-40) exposure for total RNA extracting. Then the mRNA expression levels of SNK, SPAR and NMDAR subunits NR1, NR2A and NR2B were detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: After 2-hour Abeta(1-40) exposure, we found that the expression level of SNK mRNA was up-regulated and the expression levels of SPAR, NR1, NR2A and NR2B mRNAs were down-regulated in hippocampal neurons as compared with control group (P < 0.01, P < 0.05). While with ZBPYR-containing serum pretreatment, the expression level of SNK mRNA was down-regulated and the levels of SPAR, NR1, NR2A and NR2B were up-regulated as compared with Abeta(1-40) exposure, and 2% ZBPYR-containing serum showed the best effect (P < 0.05). CONCLUSION: Abeta-induced neurotoxicity was related to SNK-SPAR pathway and NMDAR; ZBPYR-containing serum can protect neurons from Abeta-induced neurotoxicity, and this protective effect may be performed by regulating the expression of NMDAR and blocking of the SNK-SPAR pathway. PMID- 19284955 TI - Effects of Feiyanning Decoction on gene expression of nuclear factor-kappaB activated by tumor necrosis factor-alpha in lung adenocarcinoma cell line. AB - OBJECTIVE: To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on gene expression of nuclear factor-kappaB (NF-kappaB) activated by tumor necrosis factor-alpha (TNF-alpha) in lung adenocarcinoma cell line (A549). METHODS: A549 cells were incubated with rat serum containing Feiyanning at different concentrations for 24 and 48 h, respectively. Morphology of cells was observed by an inverted microscope after treatment with reagents. The cell proliferation was examined by 2-(2-methoxy-4-nitrophenyl)-3-(4 nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) assay. The expressions of NF-kappaB and inhibitor kappaBalpha (IkappaBalpha) were studied by Western blotting. NF-kappaB-dependent luciferase reporter (3 x kappaB luc) was transfected for 24 h, and the cells were treated with the reagents for 24 h, and then the transcriptional activity of NF-kappaB promoter was detected by luciferase assay. RESULTS: TNF-alpha (1 microg/L) strongly induced the expression of NF-kappaB by approximately 1.76-fold compared with the control in the nuclei of A549 cells, and the induced NF-kappaB expression was significantly suppressed by addition of Feiyanning (P < 0.01). In addition, Feiyanning inhibited the transcriptional activity of the NF-kappaB promoter. However, we observed no significant changes in IkappaBalpha expression (P > 0.05). CONCLUSION: Feiyanning Decoction can markedly inhibit human lung cancer A549 cell proliferation, which may be partly due to inhibition of NF-kappaB activation induced by TNF-alpha. It is therefore expected to be a new strategy for treating lung cancer. PMID- 19284956 TI - [Inhibitory effects of Feiyanning Decoction on proliferation, migration and tube formation of human umbilical vein endothelial cells in vitro]. AB - OBJECTIVE: To observe the inhibitory effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine for replenishing qi, nourishing essence, and diminishing stagnation by detoxification, on proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were isolated from the new-born umbilical core of human. Serum containing Feiyanning was prepared in SD rats by oral gavage of the Feiyanning Decoction for three days. The inhibitory effects of different contents of serum containing Feiyanning on the proliferation of HUVECs, human lung adenocarcinoma cell A549, and HUVECs cultured with A549-conditioned media were observed by sulforhodamine B (SRB) method; the effects on migration of HUVECs were inspected by using Boyden Chamber Transwell method, and the effects on tube formation of HUVECs were evaluated by cavity forming experiment. RESULTS: Serum containing Feiyanning at a concentration of 25%, 37.5% or 50% inhibited the proliferation of both the HUVECs and the HUVECs cultured with A549-conditioned media (P < 0.01, P < 0.05). High concentration serum containing Feiyanning inhibited the proliferation of A549 cells. Sera containing Feiyanning at concentrations of 12.5% to 37.5% inhibited the migration of HUVECs induced by 20% fetal bovine serum (P < 0.01, P < 0.05) and the formation of the primary tube of HUVECs. CONCLUSION: Feiyanning Decoction can inhibit the angiogenesis of HUVECs, which may be one of its mechanisms in inhibiting the invasion metastases of lung cancer. PMID- 19284957 TI - [Effects of Pingchuan Recipe on airway remodeling in mice with bronchial asthma]. AB - OBJECTIVE: To study the effects of Pingchuan Recipe, a compound traditional Chinese herbal medicine for treating bronchial asthma, on macrophage inflammatory protein-1alpha (MIP-1alpha) and immunoglobulin E (IgE) contents and CD86 expression in a mouse model of bronchial asthma, and to investigate the mechanism of Pingchuan Recipe in regulating airway remodeling in mice with bronchial asthma. METHODS: Fifty BALB/c mice were randomly divided into normal control group, untreated group, dexamethasone group, low-dose Pingchuan Recipe group and normal-dose Pingchuan Recipe group. Bronchial asthma in mice was induced by intra abdominal injection of mixed ovalbumin and stimulation by inhaling 5% ovalbumin. Mice in the normal control group and the untreated group did not receive treatment, but mice in the other groups were administered intragastrically with 50 g/ (kg * d) Pingchuan Recipe, 25 g/ (kg * d) Pingchuan Recipe and 0.75 mg/ (kg * d) dexamethasone, respectively. After consecutive treatment for 8 weeks, the MIP-1alpha contents in peripheral blood and bronchoalveolar lavage fluid (BALF) were measured by an avidin biotin complex-enzyme-linked immunosorbent assay (ABC ELISA) system, and the IgE contents in serum and BALF were measured by ELISA method. Direct immunofluorescence-flow cytometry was used to detect the percentages of CD86(+) and CD3(-) CD86(+) cells in peripheral blood and BALF. General histological changes of the lung tissue were observed by HE staining and collagen fiber staining 12 weeks after activation. RESULTS: Compared with the normal control group, the contents of MIP-1alpha, the percentages of CD86(+) cells and CD3(-) CD86(+) cells in peripheral blood and BALF, and the contents of IgE in serum and BALF were raised significantly (P < 0.01). The contents of MIP 1alpha, the percentages of CD86(+) cells and CD3(-) CD86(+) cells in peripheral blood and BALF, and the contents of IgE in serum and BALF in the treated groups were lower than those in the untreated group (P < 0.01). The contents of MIP 1alpha in peripheral blood and BALF and the contents of IgE in serum and BALF had no significant differences among the treated groups (P > 0.05). The percentage of CD3(-) CD86(+) cells in peripheral blood in low-dose Pingchuan Recipe group was lower than that in the dexamethasone group. Compared with the untreated group, the collagen deposition in the wall of bronchiole in the treated groups was decreased. CONCLUSION: Pingchuan Recipe can decrease the contents of MIP-1alpha, IgE and the expressions of CD86 in peripheral blood and BALF, which may be one of its mechanisms in improving chronic airway inflammation and inhibiting airway remodeling. PMID- 19284958 TI - [Dose-response patterns of Radix Glycyrrhizae in Shanghan Lun]. AB - In order to explore the dose-response patterns of Gancao (Radix Glycyrrhizae) in Shanghan Lun (Treatise on Febrile Diseases), all prescriptions containing Gancao in Shanghan Lun were analyzed by frequency and hierarchical clustering analysis. The doses of Gancao used in Shanghan Lun ranged from six zhu (Chinese unit, and one zhu is equal to 0.65 g) to four liang (Chinese unit, and one liang is equal to 15.625 g). Doses of one, two, three or four liang were commonly used. One liang Gancao as juvantia was usually matched with Mahuang (Herba Ephedrae), Xingren (Semen Armeniacae) and Guizhi (Ramulus Cinnamomi) for restricting the excessive diaphoresis of Mahuang. Two liang Gancao was often matched with some couple drugs, such as Guizhi and Shaoyao (Radix Paeoniae), Shigao (Gypsum Fibrosum) and Zhimu (Rhizoma Anemarrhenae), Fuzi (Radix Aconiti Lateralis) and Ganjiang (Rhizoma Zingiberis), for warming yang to supplement qi, nourishing yin, detoxifying Fuzi, and preventing qi impairment from heat evil. Three liang Gancao was mainly matched with Banxia (Rhizoma Pinelliae) or Renshen (Radix Ginseng) for treating middle energizer emesis. Four liang Gancao was matched with Ganjiang or tonifying herbs for invigorating vital qi and relieving spasm in deficiency syndromes with contraction, palpitation or diarrhea. Gancao is used for treating many syndromes in Shanghan Lun. It is frequently used to treat excess or heat syndromes with one or two liang in a dose and deficiency or cold syndromes with three or four liang in a dose. PMID- 19284959 TI - [Effects of penetrating acupuncture on peripheral nerve conduction velocity in patients with diabetic peripheral neuropathy: a randomized controlled trial]. PMID- 19284960 TI - [Research advances of traditional Chinese medicine in delaying skin aging]. PMID- 19284961 TI - [How to express and describe quantitative data: part two]. PMID- 19284962 TI - Comparative study on WHO Western Pacific Region and World Federation of Chinese Medicine Societies international standard terminologies on traditional medicine: an analysis of the Causes of Diseases (Part 1). PMID- 19284963 TI - Exploration on medical qigong terms in perspective of antonymy. PMID- 19284964 TI - [Major institutions conducting researches on complementary and alternative medicine in the United Kingdom: University of Exeter and University of Southampton]. PMID- 19284965 TI - Ecological fitness and strategies of adaptation of Bartonella species to their hosts and vectors. AB - Bartonella spp. are facultative intracellular bacteria that cause characteristic hostrestricted hemotropic infections in mammals and are typically transmitted by blood-sucking arthropods. In the mammalian reservoir, these bacteria initially infect a yet unrecognized primary niche, which seeds organisms into the blood stream leading to the establishment of a long-lasting intra-erythrocytic bacteremia as the hall-mark of infection. Bacterial type IV secretion systems, which are supra-molecular transporters ancestrally related to bacterial conjugation systems, represent crucial pathogenicity factors that have contributed to a radial expansion of the Bartonella lineage in nature by facilitating adaptation to unique mammalian hosts. On the molecular level, the type IV secretion system VirB/VirD4 is known to translocate a cocktail of different effector proteins into host cells, which subvert multiple cellular functions to the benefit of the infecting pathogen. Furthermore, bacterial adhesins mediate a critical, early step in the pathogenesis of the bartonellae by binding to extracellular matrix components of host cells, which leads to firm bacterial adhesion to the cell surface as a prerequisite for the efficient translocation of type IV secretion effector proteins. The best-studied adhesins in bartonellae are the orthologous trimeric autotransporter adhesins, BadA in Bartonella henselae and the Vomp family in Bartonella quintana. Genetic diversity and strain variability also appear to enhance the ability of bartonellae to invade not only specific reservoir hosts, but also accidental hosts, as shown for B. henselae. Bartonellae have been identified in many different blood-sucking arthropods, in which they are typically found to cause extracellular infections of the mid-gut epithelium. Adaptation to specific vectors and reservoirs seems to be a common strategy of bartonellae for transmission and host diversity. However, knowledge regarding arthropod specificity/restriction, the mode of transmission, and the bacterial factors involved in arthropod infection and transmission is still limited. PMID- 19284966 TI - The Marek's disease virus (MDV) protein encoded by the UL17 ortholog is essential for virus growth. AB - Marek's disease virus type 1 (MDV-1) shows a strict dependency on the direct cell to-cell spread for its propagation in cell culture. As MDV-1 shows an impaired nuclear egress in cell culture, we wished to address the characterization of capsid/tegument genes which may intervene in the maturation of intranuclear capsids. Orthologs of UL17 are present in all herpesviruses and, in all reported case, were shown to be essential for viral growth, playing a role in capsid maturation and DNA packaging. As only HSV-1 and PrV UL17 proteins have been characterized so far, we wished to examine the role of MDV-1 pUL17 in virus replication. To analyze MDV-1 UL17 gene function, we created deletion mutants or point mutated the open reading frame (ORF) to interrupt its coding phase. We established that a functional ORF UL17 is indispensable for MDV-1 growth. We chose to characterize the virally encoded protein by tagging the 729 amino-acid long protein with a repeat of the HA peptide that was fused to its C-terminus. Protein pUL17 was identified in infected cell extracts as an 82 kDa protein which localized to the nucleus, colocalizing with VP5, the major capsid protein, and VP13/14, a major tegument protein. By using green fluorescent protein fusion and HA tagged proteins expressed under the cytomegalovirus IE gene enhancer/promoter (P(CMV IE)), we showed that MDV-1 pUL17 nuclear distribution in infected cells is not an intrinsic property. Although our results strongly suggest that another viral protein retains (or relocate) pUL17 to the nucleus, we report that none of the tegument protein tested so far were able to mediate pUL17 relocation to the nucleus. PMID- 19284967 TI - Altered expression of genes related to blood-retina barrier disruption in streptozotocin-induced diabetes. AB - Disruption of the blood-retina barrier (BRB) is an early phenomenon in preclinical diabetic retinopathy (PCDR). Two vascular permeability pathways may be affected, the paracellular pathway involving endothelial cell tight junctions, and the endothelial transcellular pathway mediated by endocytotic vesicles (caveolae). The relative contribution of both pathways to vascular permeability in PCDR is unknown. We compared transcription levels in entire rat retina of genes related to these pathways between control conditions and after 6 and 12 weeks of streptozotocin-induced diabetes, as well as in bovine retinal endothelial cells (BRECs) exposed to VEGF and bovine retinal pericytes (BRPCs), using real-time quantitative RT-PCR. To confirm endothelial-specificity, immunohistochemical staining was performed in rat retina, and mRNA transcript levels were compared between BRECs and BRPCs. mRNA and protein of most paracellular transport-related genes were specifically expressed by retinal endothelial cells, whereas vesicle transport-related mRNA and proteins were present in various retinal cell types, including endothelial cells. Expression of selected endothelial cell tight junction genes and particularly that of occludin and claudin-5 was reduced in the diabetic retina and in BRECs after exposure to VEGF. Expression of 6 out of 11 vesicular transport-related genes was upregulated after induction of diabetes. Of these, only plasmalemma vesicle-associated protein (PV-1) was exclusively expressed in BRECs and not in BRPCs. PV-1 transcription was markedly induced in diabetic retina and by VEGF in BRECs. Caveolin-1 immunostaining was primarily found in the retinal vasculature, and its mRNA levels in BRECs were highly abundant and VEGF-inducible. Whereas the endothelial tight junction genes occludin and claudin-5 showed a transient downregulation, we observed long-term upregulation in diabetic retina and VEGF induced expression in BRECs of the vesicular transport-related genes caveolin-1 and PV-1. The altered gene expression profiles observed in this study suggest a transient induction of the paracellular pathway and prolonged involvement of transcellular endothelial transport mechanisms in the increased permeability of retinal capillaries in PCDR. PMID- 19284969 TI - Dynamic control of needle-free jet injection. AB - Many modern pharmaceutical therapies such as vaccines and macromolecular drugs benefit from transdermal delivery. Conventional transdermal drug delivery via hypodermic needles causes pain, non-compliance, and potential contamination. Alternative transdermal strategies that deliver drugs in a quick, reliable, painless, and inexpensive way are needed. Jet injectors, which deliver drugs through the skin using a high-speed stream of liquid propelled by compressed springs or gasses, provide a needle-free method of trandermal drug delivery. However, poor reliability as well as painful bruising and bleeding characterize these devices, due in part to the high and constant jet velocity with which drugs are delivered. Toward improved reliability and reduced pain, we have developed a jet injector capable of dynamic control of jet velocity during a single injection pulse. Using this device, we demonstrate that temporal control of jet velocity leads to independent control of penetration depth, by adjusting time at high velocity, and delivered dose, by adjusting time at low velocity, in model materials. This dynamic control of jet velocity creates the potential for better control of needle-free injections, as demonstrated through injection studies on whole ex vivo human skin samples. PMID- 19284970 TI - Regulation of expression of the tricarballylate utilization operon (tcuABC) of Salmonella enterica. AB - The tricarballylate utilization locus (tcuRABC) of Salmonella enterica serovar Typhimurium is comprised of a 3-gene operon (tcuABC) that encodes functions that allow this bacterium to use tricarballylate as a source of carbon and energy, and the tcuR gene, which encodes a putative LysR-type transcriptional regulator. In our studies, transcription of the tcuABC operon peaked at mid-log phase, and declined moderately during stationary phase. This pattern was not due to a change in the amount of TcuR in the cell, as tcuR expression did not change under the conditions tested, and TcuR did not control tcuR expression. Tricarballylate was the co-inducer. tcuABC expression was negatively affected by the cAMP receptor protein (Crp). Expression of tcuABC was one order of magnitude higher in a crp mutant strain than in the crp(+) strain; derepression of tcuABC expression was also observed in a strain lacking adenylate cyclase (Cya). At present, it is unclear whether the effect of Crp is direct or indirect. Studies with molecular mimics of tricarballylate showed that the co-inducer site restricts binding of structural mimics that contain a hydroxyl group. Two classes of TcuR constitutive variants were isolated. Class I variants responded to tricarballylate, while Class II did not. PMID- 19284971 TI - Thrombospondin-1 and CD47 regulate blood pressure and cardiac responses to vasoactive stress. AB - Nitric oxide (NO) locally regulates vascular resistance and blood pressure by modulating blood vessel tone. Thrombospondin-1 signaling via its receptor CD47 locally limits the ability of NO to relax vascular smooth muscle cells and increase regional blood flow in ischemic tissues. To determine whether thrombospondin-1 plays a broader role in central cardiovascular physiology, we examined vasoactive stress responses in mice lacking thrombospondin-1 or CD47. Mice lacking thrombospondin-1 exhibit activity-associated increases in heart rate, central diastolic and mean arterial blood pressure and a constant decrease in pulse pressure. CD47-deficient mice have normal central pulse pressure but elevated resting peripheral blood pressure. Both null mice show exaggerated decreases in peripheral blood pressure and increased cardiac output and ejection fraction in response to NO. Autonomic blockade also induces exaggerated hypotensive responses in awake thrombospondin-1 null and CD47 null mice. Both null mice exhibit a greater hypotensive response to isoflurane, and autonomic blockage under isoflurane anesthesia leads to premature death of thrombospondin-1 null mice. Conversely, the hypertensive response to epinephrine is attenuated in thrombospondin-1 null mice. Thus, the matricellular protein thrombospondin-1 and its receptor CD47 serve as acute physiological regulators of blood pressure and exert a vasopressor activity to maintain global hemodynamics under stress. PMID- 19284973 TI - Cross-complementation of Clostridium perfringens PLC and Clostridium septicum alpha-toxin mutants reveals PLC is sufficient to mediate gas gangrene. AB - Clostridium perfringens and Clostridium septicum are the most common causes of clostridial myonecrosis or gas gangrene. Although they mediate a similar disease pathology, they elaborate functionally very different alpha-toxins. We used a reciprocal complementation approach to assess the contribution of the primary toxin of each species to disease and found that C. perfringens alpha-toxin (PLC) was able to mediate the gross pathology of myonecrosis even in a C. septicum background, although it could not induce vascular leukostasis. Conversely, while C. septicum alpha-toxin restored some virulence to a C. perfringens plc mutant, it was less active than in its native background. PMID- 19284972 TI - Central regulation of the cough reflex: therapeutic implications. AB - In many species including humans, antagonists of NMDA-type glutamate receptors such as dextromethorphan, when used at sufficient doses, have been found to be relatively safe and effective antitussives. Similarly, now in five different species (guinea pigs, rabbits, cats, dogs and pigs), neurokinin receptor antagonists have also proven to be safe and effective antitussive agents. Both of these classes of drugs act centrally to prevent cough. A brief review of what is known about the central encoding of cough is presented, as are the advantages of centrally acting antitussives. Also discussed are new insights into cough and NMDA receptor signaling that may lead to the development of more effective antitussive agents with limited side effects and broad application in treating cough associated with a variety of aetiologies. PMID- 19284974 TI - Characterization of functional heterodimer partners in brain for a bHLH-PAS factor NXF. AB - NXF, a brain-specific bHLH-PAS transcription factor, can regulate the transcription of target genes forming heterodimer complexes, along with several other bHLH-PAS family members (Arnt1, Arnt2, BMAL1) in vitro. To characterize its dimerization partner protein(s) in vivo, we performed a co-immunoprecipitation analysis of whole brain extracts using anti-NXF IgG. In the protein fraction co precipitating with the NXF protein, in addition to the major precipitate of Arnt2 protein, a faint protein band of Arnt1 protein was consistently observed. The following in vitro co-precipitation analysis with recombinant proteins and yeast two-hybrid analysis confirmed the specific physical associations. Reporter gene analyses further revealed comparable levels of transcriptional activity with Arnt1:NXF and Arnt2:NXF combinations. mRNA expression for Arnt1 was found in several NXF-containing regions in brain, even an example with no Arnt2 expression. The data thus suggest that Arnt1, as well as Arnt2, could have the separate significance for NXF signaling, with Arnt1: NXF heterodimer complexes in vivo. PMID- 19284975 TI - A comparison of cortical and trabecular bone from C57 Black 6 mice using Raman spectroscopy. AB - Cortical and trabecular bone are both produced and maintained by the same cell types. At the microscopic scale they have a similar lamellar structure but at a macroscopic scale they are very different. Raman microscopy has been used to investigate compositional differences in the two bone types using bone from standard laboratory mice in physiological conditions. Clear differences were observed when complete spectra were compared by principal component analysis (PCA). Analysis of individual bands showed cortical bone to have compositional characteristics of older bone when compared with trabecular material, possibly due to the higher bone turnover traditionally reported in the trabecular compartment. PMID- 19284976 TI - Regulation of Dictyostelium morphogenesis by RapGAP3. AB - Rap1 is a key regulator of cell adhesion and cell motility in Dictyostelium. Here, we identify a Rap1-specific GAP protein (RapGAP3) and provide evidence that Rap1 signaling regulates cell-cell adhesion and cell migration within the multicellular organism. RapGAP3 mediates the deactivation of Rap1 at the late mound stage of development and plays an important role in regulating cell sorting during apical tip formation, when the anterior-posterior axis of the organism is formed, by controlling cell-cell adhesion and cell migration. The loss of RapGAP3 results in a severely altered morphogenesis of the multicellular organism at the late mound stage. Direct measurement of cell motility within the mound shows that rapGAP3(-) cells have a reduced speed of movement and, compared to wild-type cells, have a reduced motility towards the apex. rapGAP3(-) cells exhibit some increased EDTA/EGTA sensitive cell-cell adhesion at the late mound stage. RapGAP3 transiently and rapidly translocates to the cell cortex in response to chemoattractant stimulation, which is dependent on F-actin polymerization. We suggest that the altered morphogenesis and the cell-sorting defect of rapGAP3(-) cells may result in reduced directional movement of the mutant cells to the apex of the mound. PMID- 19284977 TI - Novel metabolic activity indicator in Streptococcus mutans biofilms. AB - Antimicrobial resistance of micro-organisms in biofilms requires novel strategies to evaluate the efficacy of caries preventive agents in actual biofilms. Hence we investigated fluorescence intensity (FI) in Streptococcus mutans biofilms constitutively expressing green fluorescent protein (GFP). Upon addition of glucose FI in these biofilms increased significantly to steady state levels. FI increase could be inhibited by oral care products in a dose-responsive manner. Lactic acid produced in these biofilms was measured at the end of the FI recording. A linear correlation with a coefficient of 0.96 (p<0.01) was observed between FI-increase and lactate production, irrespective of the inhibitor used. The viability of biofilm cells after chlorhexidine (CHX) titration was also examined. Reduction of FI-increase was observed at low concentrations of CHX whereas a loss in viability was only seen at high concentrations. In conclusion, GFP synthesis can be used as a metabolic activity indicator in S. mutans biofilms. PMID- 19284978 TI - Real-time PCR for the early detection and quantification of Coxiella burnetii as an alternative to the murine bioassay. AB - Real-time PCR was used to analyze archived blood from non-human primates (NHP) and fluid samples originating from a well-controlled Q fever vaccine efficacy trial. The PCR targets were the IS1111 element and the com1 gene of Coxiella burnetii. Data from that previous study were used to evaluate real-time PCR as an alternative to the use of sero-conversion by mouse bioassay for both quantification and early detection of C. burnetii bacteria. Real-time PCR and the mouse bioassay exhibited no statistical difference in quantifying the number of microorganisms delivered in the aerosol challenge dose. The presence of C. burnetii in peripheral blood of non-human primates was detected by real-time PCR as early after exposure as the mouse bioassay with results available within hours instead of weeks. This study demonstrates that real-time PCR has the ability to replace the mouse bioassay to measure dosage and monitor infection of C. burnetii in a non-human primate model. PMID- 19284979 TI - Forest age correlates with fine-scale spatial structure of Matsutake mycorrhizas. AB - Examining the fine-scale spatial structure of fungal populations can tell us much about how individual species reproduce and disperse throughout natural landscapes. Here we study the fine-scale genetic structure of Tricholoma matsutake, a prized edible and medicinal mushroom, by systematic sampling of mycorrhizas within fairy rings in 50-y-old and old-growth forests in two villages. Using single nucleotide polymorphism DNA markers we show that mycorrhizas in both forest age classes in both villages showed high levels of genotypic diversity, consistent with a reproductive life history predominated by outcrossing via basidiospore dispersal. Both the percentage of polymorphic loci within fairy rings, as well as genotype diversity were higher in old-growth compared to 50-y-old forests. Fifty-year-old forests showed significant spatial autocorrelation between pairs of mycorrhizas up to 42m, and a pattern consistent isolation-by-distance structure. Spatial patterns in old-growth forests were random. Furthermore, AMOVA analysis indicates that 11% of molecular variance in 50-y-old forests is partitioned between villages, whereas no significant variance is partitioned between villages in old-growth forests. We conclude that populations of T. matsutake in 50-y-old forests are the result of a founder effect maintained by local inoculation sources. This pattern attenuates as forests age and accumulate inocula from more distance sources. We speculate on how genetic mosaicism within T. matsutake fairy rings may structure populations within a chronosequence. Finally, we discuss how population spatial dynamics and dispersal strategy in T. matsutake contrast with other ectomycorrhizal species. PMID- 19284980 TI - Proteomic identification of extracellular proteins regulated by the Gna1 Galpha subunit in Stagonospora nodorum. AB - The fungus Stagonospora nodorum is the causal agent of stagonospora nodorum blotch (syn. leaf and glume blotch) disease of wheat. The Gna1-encoded Galpha protein is an important signal transduction component in the fungus, which is required for full pathogenicity, sporulation and extracellular depolymerase production. In this study, we sought to gain a better understanding of defects associated with the gna1 mutant by using two-dimensional gel electrophoresis to analyse the extracellular proteome for differences to the wildtype. Mass spectrometry analysis of altered abundant protein spots and peptide matching to the Stagonospora nodorum genome database have led to the identification of genes implicated in cell wall degradation, proteolysis, RNA hydrolysis and aromatic compound metabolism. In addition, quantitative RT-PCR has demonstrated that some of the encoding genes showed differential expression throughout host infection. Implications of these proteins and their corresponding genes in fungal virulence are discussed. PMID- 19284981 TI - Refolding and reconstitution of human recombinant Bax inhibitor-1 into liposomes from inclusion bodies expressed in Escherichia coli. AB - In this study, we report the simultaneous refolding and reconstitution of the recombinant Bax inhibitor-1 (BI-1) from inclusion bodies expressed in Escherichia coli. A functional assay showed that the resulting proteoliposomes responded to acidic conditions and triggered the release of entrapped Ca(2+) from liposomes. The secondary structure of the reconstituted BI-1 was also determined using circular dichroism, which revealed an increase of alpha-helix content and a decrease of random structure when exposed to acidic solutions. These conformational changes may be responsible for the proton ion-induced Ca(2+) release of BI-1. PMID- 19284982 TI - Engineering, cloning, and functional characterization of recombinant LIM mineralization protein-1 containing an N-terminal HIV-derived membrane transduction domain. AB - Short peptide sequences known as protein transduction domains have become increasingly prevalent as tools to internalize molecules that would otherwise remain extracellular. Here, we determine whether a purified recombinant mammalian intracellular osteogenic factor delivered by a HIV-derived TAT-peptide tag is indeed capable of intracellular localization in a form accessible to interaction with other proteins. We engineered and bacterially expressed a TAT-fusion-cDNA construct of a known osteogenic factor, LIM mineralization protein-1 (LMP-1) involved in the bone morphogenetic protein (BMP) pathway that has the potential to serve as an enhancer of BMP-2 efficacy. The expressed recombinant protein contains an N-terminal (His)(6)-tag, a hemagglutinin(HA)-tag, and an 11-amino acid HIV-derived TAT-membrane transduction domain and was purified to homogeneity by Sephacryl S-100 molecular exclusion and Ni(2+)-affinity chromatography. The purified TAT-LMP-1 protein was chemically labeled with fluorescein, and its time and concentration dependent entry into rabbit blood cells was monitored by flow cytometry. We demonstrate the accumulation of TAT-tagged LMP-1 both in cytoplasmic and nuclear compartments. By performing affinity pull-down assays, we confirm our earlier findings that the recombinant TAT-LMP-1, when used as molecular bait to identify the intracellular binding proteins, interacts with Smurf1, a known binding partner of LMP-1. We also show potentiation of BMP-2 activity using the purified TAT-LMP-1 in mouse muscle C2C12 cells by assaying a heterologous luciferase-reporter construct containing multiple copies of a BMP responsive sequence motif. Finally, we also confirm the biological activity of the purified TAT-LMP-1 by showing enhancement of BMP-2 induced increase of alkaline phosphatase mRNA and protein by RT-PCR and enzyme activity, respectively. PMID- 19284983 TI - Cloning, characterization and immunolocalization of two subunits of calcineurin from pearl oyster (Pinctada fucata). AB - Calcineurin (CN), consisting of catalytic subunit (CN A) and regulatory subunit (CN B), is a multifunctional protein involved in many important physiological processes. Here, we cloned two subunits of CN (Pf-CN A and Pf-CN B) from pearl oyster Pinctada fucata and reported, for the first time, its expression patterns in the developmental stages, its enzymatic activity and immunolocalization in various tissues of adult pearl oyster. The Pf-CN A was extensively localized in all the tested tissues including mantle, gonad, digestive gland, gills, adductor muscle, and foot with strong signals detected in gonad, gills, foot, and mantle. Importantly, Pf-CN A was mainly found in the inner epithelial cells of the basal periostracal groove and lateral surface of the inner mantle fold, in which organic macromolecules used for periostracum formation and shell construction are secreted, respectively. In gill, the strong signals were distributed in the epithelial cells of the branchial filaments and the base of gill filaments. All the results suggested that Pf-CN may participate in the development of the pearl oyster and function in many ways in various physiological activities, especially in the shell formation. Our observations could provide some important clues to further understanding of the functions of CN in the oyster. PMID- 19284984 TI - Another patient with a de novo deletion further delineates the 2q33.1 microdeletion syndrome. AB - A male patient, who had intra-uterine growth retardation, a low birth weight and hypotonia due to a chromosome 2q33.1 deletion, is described. At the age of 20 years, he displays short stature, microcephaly, a high forehead, microstomia, large teeth and is hypertonic. He is severely mentally retarded, has not developed speech, is hyperactive, anxious and at times aggressive. Full tiling array showed a de novo 14 Mb deletion at chromosome region 2q32.3q33.2, further delineating the 2q33.1 microdeletion syndrome. PMID- 19284985 TI - Evolution of gene regulatory network architectures: examples of subcircuit conservation and plasticity between classes of echinoderms. AB - Developmental gene regulatory networks (GRNs) explain how regulatory states are established in particular cells during development and how these states then determine the final form of the embryo. Evolutionary changes to the sequence of the genome will direct reorganization of GRN architectures, which in turn will lead to the alteration of developmental programs. A comparison of GRN architectures must consequently reveal the molecular basis for the evolution of developmental programs among different organisms. This review highlights some of the important findings that have emerged from the most extensive direct comparison of GRN architectures to date. Comparison of the orthologous GRNs for endomesodermal specification in the sea urchin and sea star, provides examples of several discrete, functional GRN subcircuits and shows that they are subject to diverse selective pressures. This demonstrates that different regulatory linkages may be more or less amenable to evolutionary change. One of the more surprising findings from this comparison is that GRN-level functions may be maintained while the factors performing the functions have changed, suggesting that GRNs have a high capacity for compensatory changes involving transcription factor binding to cis regulatory modules. PMID- 19284986 TI - Poly(C)-binding proteins as transcriptional regulators of gene expression. AB - Poly(C)-binding proteins (PCBPs) are generally known as RNA-binding proteins that interact in a sequence-specific fashion with single-stranded poly(C). They can be divided into two groups: hnRNP K and PCBP1-4. These proteins are involved mainly in various posttranscriptional regulations (e.g., mRNA stabilization or translational activation/silencing). In this review, we summarize and discuss how PCBPs act as transcriptional regulators by binding to specific elements in gene promoters that interact with the RNA polymerase II transcription machinery. Transcriptional regulation of PCBPs might itself be regulated by their localization within the cell. For example, activation by p21-activated kinase 1 induces increased nuclear retention of PCBP1, as well as increased promoter activity. PCBPs can function as a signal-dependent and coordinated regulator of transcription in eukaryotic cells. We address the molecular mechanisms by which PCBPs binding to single- and double-stranded DNA mediates gene expression. PMID- 19284987 TI - In vivo profile of the human leukocyte microRNA response to endotoxemia. AB - To gain insight into microRNAs (miRNAs) involved in the regulation of the human innate immune response, we screened for differentially expressed miRNAs in circulating leukocytes in an in vivo model of acute inflammation triggered by Escherichia coli lipopolysaccharide (LPS) infusion. Leukocyte RNA was isolated from venous blood samples obtained from healthy male volunteers before and 4h after LPS-infusion. After fluorescence labeling, RNA samples were hybridized to microarrays containing capture probes for measuring the abundance of more than 600 human miRNAs. Target genes were predicted for differentially expressed miRNAs and then compared to changes in genome-wide expression levels, which had been established in a previous study. Data analysis revealed that five miRNAs consistently responded to LPS-infusion, four of which were down-regulated (miR 146b, miR-150, miR-342, and let-7g) and one was up-regulated (miR-143). The miR 150 and mir-342 response was confirmed by real-time PCR. By correlating to measured LPS-induced changes of the leukocyte transcriptome, we next searched for predicted target genes, whose stability might be under (co-) control by these miRNAs. We found that the rapid transcriptional activation during acute inflammation of select genes, such as the gene encoding interleukin-1 receptor associated kinase 2 (IRAK2) might be facilitated by decreased levels of LPS responsive miRNAs. The increased level of miR-143 might be associated with the pronounced down-regulation of the B-cell CLL/lymphoma 2 (BCL2) gene expression during LPS endotoxemia, and could further be involved in the translational silencing of several other predicted inflammation-related target genes. This is the first in vivo study to demonstrate relative abundance of miRNA levels in peripheral blood leukocytes during acute LPS-induced inflammation. The miRNAs and their potential target genes identified herein contribute to the understanding of the complex transcriptional program of innate immunity initiated by pathogens. PMID- 19284988 TI - Ptpcd-1 is a novel cell cycle related phosphatase that regulates centriole duplication and cytokinesis. AB - Proper progression of mitosis requires spatio-temporal regulation of protein phosphorylation by orchestrated activities of kinases and phosphatases. Although many kinases, such as Aurora kinases, polo-like kinases (Plks), and cyclin B-Cdk1 are relatively well characterized in the context of their physiological functions at mitosis and regulation of their enzymatic activities during mitotic progression, phosphatases involved are largely unknown. Here we identified a novel protein tyrosine phosphatase containing domain 1 (Ptpcd 1) as a mitotic phosphatase, which shares sequence homology to Cdc14. Immunofluorescence studies revealed that Ptpcd1 partially colocalized with gamma-tubulin, an archetypical centrosomal marker. Overexpression of this phosphatase prevented unscheduled centrosomal amplification in hydroxyurea arrested U2OS cells. Intriguingly, Ptpcd 1-associated and colocalized with polo-like kinase 1(Plk1). Hence, overexpression of Ptpcd1 rescued prometaphase arrest of Plk-1 depleted cells, but resulted in aberrant cytokinesis as did as Plk1 overexpression. These results suggested that Ptpcd1 is involved in centrosomal duplication and cytokinesis. PMID- 19284989 TI - Crystal structures of respiratory pathogen neuraminidases. AB - Currently there is pressing need to develop novel therapeutic agents for the treatment of infections by the human respiratory pathogens Pseudomonas aeruginosa and Streptococcus pneumoniae. The neuraminidases of these pathogens are important for host colonization in animal models of infection and are attractive targets for drug discovery. To aid in the development of inhibitors against these neuraminidases, we have determined the crystal structures of the P. aeruginosa enzyme NanPs and S. pneumoniae enzyme NanA at 1.6 and 1.7A resolution, respectively. In situ proteolysis with trypsin was essential for the crystallization of our recombinant NanA. The active site regions of the two enzymes are strikingly different. NanA contains a deep pocket that is similar to that in canonical neuraminidases, while the NanPs active site is much more open. The comparative studies suggest that NanPs may not be a classical neuraminidase, and may have distinct natural substrates and physiological functions. This work represents an important step in the development of drugs to prevent respiratory tract colonization by these two pathogens. PMID- 19284990 TI - Expression and localization of regenerating gene I in a rat liver regeneration model. AB - Regenerating gene (Reg) I has been identified as a regenerative/proliferative factor for pancreatic islet cells. We examined Reg I expression in the regenerating liver of a rat model that had been administered 2 acetylaminofluorene and treated with 70% partial hepatectomy (2-AAF/PH model), where hepatocyte and cholangiocyte proliferation was suppressed and the hepatic stem cells and/or hepatic progenitor cells were activated. In a detailed time course study of activation of hepatic stem cells in the 2-AAF/PH model, utilizing immunofluorescence staining with antibodies of Reg I and other cell-type-specific markers, we found that Reg I-expressing cells are present in the bile ductules and increased during regeneration. Reg I-expressing cells were colocalized with CK19, OV6, and AFP. These results demonstrate that Reg I is significantly upregulated in the liver of the 2-AAF/PH rat model, accompanied by the formation of bile ductules during liver regeneration. PMID- 19284991 TI - Smad4 restoration leads to a suppression of Wnt/beta-catenin signaling activity and migration capacity in human colon carcinoma cells. AB - Recent studies have reported that Smad4 has a TGF-beta-independent function as a tumor suppressor in cooperating with beta-catenin/Lef to regulate target gene expression. The objective of this research was to study the role of Smad4 in colon cancer migration and its potential mechanism. In our study, stable colon cancer cells that have increased Smad4 expression were created using a eukaryotic vector containing Smad4-pcDNA3.1(+). Smad4 restoration directly suppressed the Wnt/beta-catenin signal activity in SW480 cells by down-regulating beta-catenin expression and altering its localization from cytoplasm and nucleus to the plasma membrane. Up-regulation of Smad4 also increased the expression of E-cadherin and decreased the transcriptional activity of beta-catenin/Tcf target genes, such as claudin-1 and MMP-7. SW480 cells with increased Smad4 expression had decreased in vitro cell migration. The data suggested that restoration of Smad4 in Smad4 deficient cells may provide a potential therapeutic strategy for intervention of colon cancer migration and metastasis. PMID- 19284992 TI - Glycine-rich region regulates cysteine-rich protein 1 binding to actin cytoskeleton. AB - Cysteine-rich protein 1 (CRP1) has a unique structure with two well separated LIM domains, each followed by a glycine-rich region. Although CRP1 has been shown to interact with actin-binding proteins and actin filaments, the mechanism regulating localization to the actin cytoskeleton in cells is not clear. Experiments using truncated forms showed that the first LIM domain and glycine rich region are necessary for CRP1 bundling of actin filaments and localization to the actin cytoskeleton. Furthermore, domain swapping experiments replacing the first glycine-rich region with the second resulted in the loss of CRP1 bundling activity and localization to the actin cytoskeleton, identifying seven critical amino acid residues. These results highlight the importance of the first glycine rich region for CRP1 bundling activity and localization to the actin cytoskeleton. In addition, this work identifies the first LIM domain and glycine rich region as a distinct actin filament bundling module. PMID- 19284993 TI - Defective regulation of the ryanodine receptor induces hypertrophy in cardiomyocytes. AB - Recent studies on cardiac hypertrophy animal model suggest that inter-domain interactions within the ryanodine receptor (RyR2) become defective concomitant with the development of hypertrophy (e.g. de-stabilization of the interaction between N-terminal and central domains of RyR2; T. Oda, M. Yano, T. Yamamoto, T. Tokuhisa, S. Okuda, M. Doi, T. Ohkusa, Y. Ikeda, S. Kobayashi, N. Ikemoto, M. Matsuzaki, Defective regulation of inter-domain interactions within the ryanodine receptor plays a key role in the pathogenesis of heart failure, Circulation 111 (2005) 3400-3410). To determine if de-stabilization of the inter-domain interaction in fact causes hypertrophy, we introduced DPc10 (a peptide corresponding to the G(2460)-P(2495) region of RyR2, which is known to de stabilize the N-terminal/central domain interaction) into rat neonatal cardiomyocytes by mediation of peptide carrier BioPORTER. After incubation for 24h the peptide induced hypertrophy, as evidenced by significant increase in cell size and [(3)H]leucine uptake. K201 or dantrolene, the reagents known to correct the de-stabilized inter-domain interaction to a normal mode, prevented the DPc10 induced hypertrophy. These results suggest that disruption of the normal N terminal/central inter-domain interaction within the RyR2 is a causative mechanism of cardiomyocyte hypertrophy. PMID- 19284994 TI - Crystal structure of the human tRNAGly microhelix isoacceptor G9990 at 1.18A resolution. AB - The tRNA(Gly)/Glycyl-tRNA synthetase system belongs to the so called 'class II' in which tRNA identity elements consist of relative few and simple motifs, as compared to 'class I' where the tRNA determinants are more complicated and spread over different parts of the tRNA, mostly including the anticodon. The determinants from 'class II' although, are located in the aminoacyl stem and sometimes include the discriminator base. There exist predominant structure differences for the Glycyl-tRNA-synthetases and for the tRNA(Gly) identity elements comparing eucaryotic/archaebacterial and eubacterial systems. We focus on comparative X-ray structure analysis of tRNA(Gly) acceptor stem microhelices from different organisms. Here, we report the X-ray structure of the human tRNA(Gly) microhelix isoacceptor G9990 at 1.18A resolution. Superposition experiments to another human tRNA(Gly) microhelix and a detailed comparison of the RNA hydration patterns show a great number of water molecules with identical positions in both RNAs. This is the first structure comparison of hydration layers from two isoacceptor tRNA microhelices with a naturally occurring base pair exchange. PMID- 19284995 TI - S100A13-C2A binary complex structure-a key component in the acidic fibroblast growth factor for the non-classical pathway. AB - Fibroblast growth factors (FGFs) are key regulators of cell proliferation, differentiation, tumor-induced angiogenesis and migration. FGFs are essential for early embryonic development, organ formation and angiogenesis. They play important roles in tumor formation, inflammation, wound healing and restenosis. The biological effects of FGFs are mediated through the activation of the four transmembrane phosphotyrosine kinase receptors (FGFRs) in the presence of heparin sulfate proteoglycans (HSPGs) and therefore require the release of FGFs into the extracellular space. However, FGF-1 lacks the signal peptide required for the releasing of these proteins through the classical endoplasmic reticulum (ER) Golgi secretary pathway. Maciag et al. demonstrated that FGF-1 is exported through a non-classical release pathway involving the formation of a specific multiprotein complex [M. Landriscina, R. Soldi, C. Bagala, I. Micucci, S. Bellum, F. Tarantini, I. Prudovsky, T. Maciag, S100A13 participates in the release of fibroblast growth factor 1 in response to heat shock in vitro, J. Biol. Chem. 276 (2001) 22544-22552; C.M. Carreira, T.M. LaVallee, F. Tarantini, A. Jackson, J.T. Lathrop, B. Hampton, W.H. Burgess, T. Maciag, S100A13 is involved in the regulation of fibroblast growth factor-1 and p40 synaptotagmin-1 release in vitro, J. Biol. Chem. 273 (1998) 22224-22231; T.M. LaValle, F. Tarantini, S. Gamble, C.M. Carreira, A. Jackson, T. Maciag, Synaptotagmin-1 is required for fibroblast growth factor-1 release, J. Biol. Chem. 273 (1998) 22217-22223; C. Bagala, V. Kolev, A. Mandinova, R. Soldi, C. Mouta, I. Graziani, I, Prudovsky, T. Maciag, The alternative translation of synaptotagmin 1 mediates the non-classical release of FGF1, Biochem. Biophys. Res. Commun. 310 (2003) 1041-1047]. The protein constituents of this complex include FGF-1, S100A13 (a Ca(2+)-binding protein), and the p40 form of synaptotagmin 1 (Syt1). To understand the molecular events in the FGF-1 releasing pathway, we have studied the interactions of S100A13 with C2A by (1)H-(15)N HSQC titration and 3D-filtered NOESY experiments. We characterized the binary complex structure of S100A13-C2A by using a variety of multi-dimensional NMR experiments. This complex acts as a template for FGF-1 dimerization and multiprotein complex formation. PMID- 19284996 TI - Both foxj1a and foxj1b are implicated in left-right asymmetric development in zebrafish embryos. AB - The HNF-3/HFH-4/Foxj1, a transcription factor, has been reported to be involved in systemic autoimmunity and cilia genesis in vertebrates. The zebrafish genome expressed two paralogous foxj1 genes, foxj1a and foxj1b. In this study, we demonstrate that down-regulation of either foxj1a or foxj1b by injecting antisense morpholino at the one-cell stage results in randomized expression of the early left-right (LR) asymmetric markers lefty2, southpaw, pitx2c and the later internal organ markers tpm4-tv1, cmlc2, cp in zebrafish embryos. Overexpression of foxj1a and foxj1b by injecting synthetic mRNAs also disrupts normal LR asymmetries. These data indicate that the two foxj1 genes are required for normal laterality development in zebrafish embryos. In contrast to foxj1b knockdown exclusively in dorsal forerunner cells (DFCs) that has little effect on laterality, foxj1a knockdown in DFCs randomizes the LR patterns of the markers. Thus, foxj1a regulates asymmetric development through DFCs in a cell-autonomous fashion but foxj1b functions indirectly. PMID- 19284997 TI - Human beta-2 microglobulin W60V mutant structure: Implications for stability and amyloid aggregation. AB - Beta-2 microglobulin (?2m) is the light chain of class I major histocompatibility complex (MHC-I). Beta2m is an intrinsically amyloidogenic protein that can assemble into amyloid fibrils in a concentration dependent manner. Beta2m is accumulated in serum of haemodialysed patients, and deposited in the skeletal joints, causing dialysis related amyloidosis. Recent reports suggested that the loop comprised between beta2m strands D and E is crucial for protein stability and for beta2m propensity to aggregate as cross-beta structured fibrils. In particular, the role of Trp60 for beta2m stability has been highlighted by showing that the Trp60-->Gly beta2m mutant is more thermo-stable and less prone to aggregation than the wild type protein. On the contrary the Asp59-->Pro beta2m mutant shows lower Tm and stronger tendency to fibril aggregation. To further analyse such properties, the Trp60-->Val beta2m mutant has been expressed and purified; the propensity to fibrillar aggregation and the folding stability have been assessed, and the X-ray crystal structure determined to 1.8A resolution. The W60V mutant structural features are discussed, focusing on the roles of the DE loop and of residue 60 in relation to ?2m structure and its amyloid aggregation trends. PMID- 19284998 TI - Novel mutations of ABCC6 gene in Japanese patients with Angioid streaks. AB - Angioid streaks (AS) are eye abnormalities caused by breaks in Bruch's membrane. The condition is often associated with pseudoxanthoma elasticum (PXE). The ATP binding cassette, sub-family C (CFTR/MRP), member 6 (ABCC6) is reported to be the causal gene for PXE, although there have been no reports on whether the ABCC6 gene is the causal gene for AS. The aims of this study are to isolate the causal mutations for AS using a haplotype-based case-control study. We genotyped 54 Japanese AS patients and 150 controls for 5 single-nucleotide polymorphisms (SNPs). A simple association study using each SNP and a haplotype-based case control study were performed. Twelve patients with special haplotypes for AS were selected, and were then subjected to gene sequencing. Six variants were successfully identified as causal mutations for AS (p.R419Q, p.E422K, c.2542delG, Del_Exon23, c.3774-3775insC and p.E1427K), and 4 of these were novel. This method can be applied to both identifying susceptibility variants of multifactorial diseases and isolating mutations in single-gene diseases. PMID- 19284999 TI - Development of a novel fluorescent substrate for Autolysin E, a bacterial type II amidase. AB - The bifunctional Autolysin E from Staphylococcus epidermidis, contains a Zn(2+) dependent N-acetylmuramoyl-L-alanine amidase AmiE (EC 3.5.1.28). This enzyme hydrolyzes the amide bond between the carbohydrate chain and the peptide stem of bacterial peptidoglycan. Since peptidoglycan is the mayor component of bacterial cell walls, type II amidases like Autolysin E play an essential role in the bacterial life cycle. Therefore bacterial amidases are appropriate drug targets in the development of antibiotics. The drug discovery process relies on sensitive enzyme assay systems to test possible lead candidates for enzyme inhibition. However, specific determination of bacterial amidase activity is complicated because a simple and accurate enzyme assay is currently unavailable. In this study we developed a sensitive fluorescent substrate for the type II amidase Autolysin E from S. epidermidis, which is suitable for quantifying amidase activity in a high throughput compatible fashion. Using derivatives of the substrate Mca-Ala-D-isoGln-Lys(Dnp)-D-Ala-Arg-OH, we were further able to characterize the amidase substrate specificity of Autolysin E. PMID- 19285000 TI - Expression and role of TTF-1 in the rat suprachiasmatic nucleus. AB - We have previously reported that TTF-1, a homeodomain-containing transcription factor, regulates circadian rhythm of pituitary adenylate cyclase-activating polypeptide gene expression in the rat hypothalamus. In this study we found that TTF-1 mRNA was specifically expressed in the rat suprachiasmatic nucleus (SCN) and colocalized with Period 2 (Per2), a circadian feedback loop controller. Interaction between TTF-1 and Per1 and Per2 was demonstrated by immunoprecipitation and immunoblot assays. Moreover, TTF-1 and Per proteins additively stimulated a transcriptional activity of angiotensinogen (AoGen) gene. TTF-1 also activated in vitro rhythm of AoGen transcription determined by secretary alkaline phosphatase (SEAP) reporter system in the NIH3T3 cells. These results suggest that TTF-1 plays a role in the circadian rhythm regulation of the AoGen gene expression via interacting with Per proteins in the rat SCN. PMID- 19285001 TI - Conformational detection of prion protein with biarsenical labeling and FlAsH fluorescence. AB - Prion diseases are associated with the misfolding of the host-encoded cellular prion protein (PrP(C)) into a disease associated form (PrP(Sc)). Recombinant PrP can be refolded into either an alpha-helical rich conformation (alpha-PrP) resembling PrP(C) or a beta-sheet rich, protease resistant form similar to PrP(Sc). Here, we generated tetracysteine tagged recombinant PrP, folded this into alpha- or beta-PrP and determined the levels of FlAsH fluorescence. Insertion of the tetracysteine tag at three different sites within the 91-111 epitope readily distinguished beta-PrP from alpha-PrP upon FlAsH labeling. Labelling of tetracysteine tagged PrP in the alpha-helical form showed minimal fluorescence, whereas labeling of tagged PrP in the beta-sheet form showed high fluorescence indicating that this region is exposed upon conversion. This highlights a region of PrP that can be implicated in the development of diagnostics and is a novel, protease free mechanism for distinguishing PrP(Sc) from PrP(C). This technique may also be applied to any protein that undergoes conformational change and/or misfolding such as those involved in other neurodegenerative disorders including Alzheimer's, Huntington's and Parkinson's diseases. PMID- 19285002 TI - O-GlcNAc inhibits interaction between Sp1 and Elf-1 transcription factors. AB - The novel protein modification, O-linked N-acetylglucosamine (O-GlcNAc), plays an important role in various aspects of cell regulation. Although most of nuclear transcription regulatory factors are modified by O-GlcNAc, O-GlcNAc effects on transcription remain largely undefined yet. In this study, we show that O-GlcNAc inhibits a physical interaction between Sp1 and Elf-1 transcription factors, and negatively regulates transcription of placenta and embryonic expression oncofetal protein gene (Pem). These findings suggest that O-GlcNAc inhibits Sp1-mediated gene transcription possibly by interrupting Sp1 interaction with its cooperative factor. PMID- 19285003 TI - Micropatterned co-cultures of T-lymphocytes and epithelial cells as a model of mucosal immune system. AB - Gut-associated lymphoid tissue is a major target and reservoir of human immunodeficiency virus (HIV)-infected T-cells. Our studies seek to recapitulate, in vitro, interactions between HIV-infected T-lymphocytes and intestinal epithelial cells in order to investigate the mechanisms underlying the disruption of normal epithelial cell and barrier function. Here, we describe a novel approach for creating co-cultures of healthy or HIV-infected T-lymphocytes (Jurkat) and human intestinal epithelial (HT-29) cells where both cell types are positioned on the same surface in a price spatial configuration (micropattern). This co-culture method simplified observation/monitoring of the two cell types and was particularly suited for laser microdissection-based retrieval of the desired cells for downstream gene expressions studies. DNA microarray analysis of epithelial cells retrieved from co-cultures with HIV-1-infected vs. uninfected Jurkat cells revealed that epithelial cells from HIV-infected co-cultures exhibited gene expression patterns consistent with disruption of epithelial barrier formation. Overall, the micropatterned co-culture system described here is envisioned as a valuable new tool for delineating how HIV and other infections contribute to dysfunction of mucosal epithelium. PMID- 19285004 TI - Stability and solubility of proteins from extremophiles. AB - Charges are important for hyperthermophile protein structure and function. However, the number of charges and their predicted contributions to folded state stability are not correlated, implying that more charge does not imply greater stability. The charge properties that distinguish hyperthermophile proteins also differentiate psychrophile proteins from mesophile proteins, but in the opposite direction and to a smaller extent. We conclude that charge number relates to solubility, whereas protein stability is determined by charge location. Most other structural properties are poorly separated over the ambient temperature range, apart from the burial of certain amino acids. Of particular interest are large non-polar sidechains that tend to increased exposure in proteins evolved to function at higher temperatures. Looking at tryptophan in more detail, this increase is often located close to the termini of secondary structure elements, and is discussed in terms of a novel potential role in protein thermostabilisation. PMID- 19285005 TI - Action of N-acylated ambroxol derivatives on secretion of chloride ions in human airway epithelia. AB - We report the effects of new N-acylated ambroxol derivatives (TEI-588a, TEI-588b, TEI-589a, TEI-589b, TEI-602a and TEI-602b: a, aromatic amine-acylated derivative; b, aliphatic amine-acylated derivative) induced from ambroxol (a mucolytic agent to treat human lung diseases) on Cl(-) secretion in human submucosal serous Calu 3 cells under a Na(+)/K(+)/2Cl(-) cotransporter-1 (NKCC1)-mediated hyper secreting condition. TEI-589a, TEI-589b and TEI-602a diminished hyper-secretion of Cl(-) by diminishing the activity of NKCC1 without blockade of apical Cl(-) channel (TEI-589a>TEI-602a>TEI-589b), while any other tested compounds including ambroxol had no effects on Cl(-) secretion. These indicate that the inhibitory action of an aromatic amine-acylated derivative on Cl(-) secretion is stronger that that of an aliphatic amine-acylated derivative, and that 3-(2,5 dimethyl)furoyl group has a strong action in inhibition of Cl(-) secretion than cyclopropanoyl group. We here indicate that TEI-589a, TEI-589b and TEI-602a reduce hyper-secretion to an appropriate level in the airway, providing a possibility that the compound can be an effective drug in airway obstructive diseases including COPD by reducing the airway resistance under a hyper-secreting condition. PMID- 19285006 TI - A direct spectrophotometric gamma-glutamyltransferase inhibitor screening assay targeting the hydrolysis-only mode. AB - Gamma-glutamyltransferase (GGT, E.C. 2.3.2.2) catalyzes the hydrolysis and transpeptidation of extracellular glutathione. Due to its central role in maintaining mammalian glutathione homeostasis, GGT is now believed to be a valuable drug target for a variety of life-threatening diseases, such as cancer. Unfortunately, however, effective tools for screening GGT inhibitors are still lacking. We report here the synthesis and evaluation of an alpha-phenylthio containing glutathione peptide mimic that eliminates thiophenol upon GGT catalyzed hydrolysis of the gamma-glutamyl peptide bond. The concurrent, real time spectrophotometric quantification of the released thiophenol using Ellman's reagent creates a GGT assay format that is simple, robust, and highly sensitive. The versatility of the assay has been demonstrated by its application to the kinetic characterization of equine kidney GGT, and enzyme inhibition assays. The ability of the glutathione mimic to behave as an excellent donor substrate (exhibiting Michaelis-Menten kinetics with a K(m) of 11.3+/-0.5 microM and a k(cat) of 90.1+/-0.8 nmol mg(-1)min(-1)), coupled to the assay's ability to study the hydrolysis-only mode of the GGT-catalyzed reaction, make our approach amenable to high-throughput drug screening platforms. PMID- 19285007 TI - OsDEG10 encoding a small RNA-binding protein is involved in abiotic stress signaling. AB - Excessive light can be harmful to photosynthetic apparatus since it causes photoinhibition and photooxidation, and plants often encounter hypoxic or anoxic environments when they become submerged by heavy rain or an ensuing flood. In this study, Oryza sativa Differentially Expressed Genes (OsDEGs) from rice under photooxidation and anoxia conditions were isolated using DD-PCR. Among them, OsDEG10 is predicted to encode a small RNA-binding protein (RBP) and the transcript levels of OsDEG10 strongly increased under most of abiotic stress treatments such as high light, anoxia, NaCl, ABA, MV and cold. However, the transcript levels of two rice OsDEG10 homologs were not changed under those treatments. OsDEG10 RNAi transgenic plants were more sensitive to high light and cold stresses compared to wild-type plants. Our results suggest that OsDEG10 is a small RBP involved in the response to various abiotic stresses. PMID- 19285008 TI - Liquid-chromatographic and mass-spectrometric identification of lens proteins using microwave-assisted digestion with trypsin-immobilized magnetic nanoparticles. AB - We used a newly developed method combining trypsin-immobilized magnetic nanoparticles (TIMNs) and microwave-assisted protein digestion to study the proteins of human lens tissue. The digested proteins were identified by liquid chromatography and mass spectrometry. The lens proteins were digested under optimized conditions (digestion time 1 min, microwave power 400 W, trypsin-to protein ratio 1:5) determined using bovine serum albumin as the standard protein, before liquid-chromatographic and mass-spectrometric analysis. Twenty-six proteins were identified with the new digestion method compared with 11 proteins identified with traditional in-solution digestion (12h). gamma-Crystallin, beta crystallin, and superoxide dismutase 1 proteins, identified with the microwave assisted method but not the traditional method, are related to cataract development according to some studies. The TIMNs were easily separated from the digestion products. This new digestion method could prove extremely useful for large-scale proteomic analyses. PMID- 19285009 TI - Identification and characterization of stem cell-specific transcription of PSF1 in spermatogenesis. AB - PSF1 is an evolutionarily conserved DNA replication factor, which forms the GINS complex with PSF2, PSF3, and SLD5. The mouse PSF1 homolog has been identified from a stem cell-specific cDNA library. To investigate its transcriptional regulatory mechanisms during differentiation, we studied PSF1 mRNA expression in testis and characterized its promoter. No canonical TATA or CAAT boxes could be found in the PSF1 5'-flanking region, whereas several consensus AML1, GATA, and Sry putative binding sequences are predicted within 5 kb of the putative transcription start site. In addition, binding sites for oncoproteins such as Myb and Ets were also found in the promoter. In testis, various PSF1 gene transcription initiation sites are present and short transcripts encoding two novel isoforms, PSF1b and 1c, were found. However, spermatogonium stem cells specifically express transcripts for PSF1a. These data suggest that PSF1 is tightly regulated at the transcriptional level in stem cells. PMID- 19285010 TI - Troglitazone increases expression of E-cadherin and claudin 4 in human pancreatic cancer cells. AB - We examined the effects of troglitazone on expression of E-cadherin and claudin 4 in human pancreatic cancer cells. Troglitazone dose-dependently increased expression of E-cadherin and claudin 4 mRNA and protein in PK-1 cells. Snail, Slug and ZEB1, mRNAs were not changed by troglitazone, indicating that these three transcriptional repressors would not play a role in the induction of E cadherin by troglitazone. GW9662, a PPARgamma antagonist, failed to block the increased expression of E-cadherin or claudin 4 mRNA, suggesting a PPARgamma independent pathway. A MEK inhibitor, U0126, increased E-cadherin or claudin 4 mRNA and protein expression, and potently inhibited cell invasion. Because troglitazone down-regulates MEK-ERK signaling and inhibit cell invasion in PK-1 as shown in our previous study, these results suggest that troglitazone increases expression of E-cadherin and claudin 4 possibly through inhibition of MEK-ERK signaling in pancreatic cancer cells, which might be involved in the troglitazone induced inhibition of cell invasive activity. PMID- 19285011 TI - Presence of alpha-smooth muscle actin-positive endothelial cells in the luminal surface of adult aorta. AB - Alpha-smooth muscle actin-positive endothelial cells have not been found in adult aortic endothelium except valve leaflets. Here, using en face immunostaining method, we identified alpha-smooth muscle actin-positive endothelial cells in the luminal surface of rat, mouse and human thoracic aortas. These cells express both endothelial markers and definite smooth muscle cell markers and were only occasionally observed in thoracic aorta of wild type mice and rats. Their density did not increase with aging. Given that alpha-smooth muscle actin-positive endothelial cells express low level of vascular endothelial-cadherin that is important for the maintenance of cell contact, these cells were frequently detected in the thoracic aorta of 5-week-old apolipoprotein-E deficient mice. In 20- to 24-week-old apolipoprotein-E deficient mice, marked accumulation of alpha smooth muscle actin-positive endothelial cells was observed especially in the luminal surface of atheromatous plaques. Our findings indicate the existence of alpha-smooth muscle actin-positive endothelial cells in adult aortic endothelium and the possible association with progression of atherosclerosis. PMID- 19285012 TI - Novel high-throughput screening system for identifying STAT3-SH2 antagonists. AB - Constitutive activation of the oncogenic transcription factor STAT3 frequently occurs in various human malignancies. STAT3 activation involves dimerization via intermolecular pTyr-SH2 interaction. Thus, antagonizing this interaction is a feasible approach to inhibit STAT3 activation for cancer therapy. In order to identify selective STAT3 inhibitors, we developed a biochemical HTS system based on AlphaScreen technology, which measures the abilities of test compounds to antagonize pTyr-SH2 interactions. We screened our chemical libraries using this system and identified 5,15-diphenylporphyrin (5,15-DPP) as a selective STAT3-SH2 antagonist. Selective inhibition of STAT3 nuclear translocation and DNA biding activity was observed in cells treated with 5,15-DPP. IL-6-dependent dimerization of STAT3, c-myc promoter binding and c-myc protein expression were all suppressed by 5,15-DPP, whereas no decrement in either expression or phosphorylation level of STAT3 was observed. Thus, the HTS assay system represented herein may be useful for identifying novel STAT3-SH2 antagonists. PMID- 19285013 TI - Periodate-resistant carbohydrate epitopes recognized by IgG and IgE antibodies from some of the immunized mice and patients with allergy. AB - The beta(1,2)-xylose- and/or alpha(1,3)-fucose-containing cross-reactive carbohydrate determinants (CCDs) are present in various plant and insect N glycans, and have been attracted as potential antigens in IgE-mediated allergies and immunologically undesired post-translational products on some recombinant therapeutic proteins. By using ELISA and immunoblotting, CCDs-specific IgG and IgE antibodies from some, but not all, of mice and humans were found to fully retain their binding activity after a typical periodate-treatment to CCDs, which did cause the CCDs' antigenic activity to those from the other mice and rabbits to disappear almost completely. Furthermore, the mouse IgE recognizing the periodate-resistant CCDs induced the CCDs/IgE-dependent degranulation of rat basophilic RBL-2H3 cells. These findings indicate that in some cases CCDs include those dependent of the core trisaccharide more strongly than the terminal xylose and fucose, which might have been screened out in the CCDs analyses based on the loss of antibody-binding by the periodate-treatment. PMID- 19285014 TI - The serine 2481-autophosphorylated form of mammalian Target Of Rapamycin (mTOR) is localized to midzone and midbody in dividing cancer cells. AB - Using a high-resolution, automated confocal high-content imaging system, we investigated the sub-cellular localization of the Serine 2481-autophosphorylated form of mTOR (PP-mTOR(Ser2481)) during mitosis and cytokinesis in human cancer cells. PP-mTOR(Ser2481) exhibited a punctate nuclear distribution in interphase cancer cells, with the number of PP-mTOR(Ser2481) nuclear speckles positively relating with the proliferative capacity of cancer cells. PP-mTOR(Ser2481) expression dynamically rearranged within the cytoplasm in a close association near and between separating chromosomes during early stages of mitosis. Towards the end of anaphase and in telophase, PP-mTOR(Ser2481) drastically focused on the midzone and ultimately in the centre of the midbody at the presumptive cleavage furrow. In cells at cytokinesis, PP-mTOR(Ser2481) appeared as a doublet facing each other at the apical ends of two daughter cells. Three-dimensional analysis confirmed that PP-mTOR(Ser2481) positioned at a ring structure wrapped round by microtubule bundles to connect daughter cells. These results reveal for the first time that PP-mTOR(Ser2481) may be unexpectedly involved in the terminal stages of cytokinesis. PMID- 19285015 TI - Resveratrol inhibits the expression of SREBP1 in cell model of steatosis via Sirt1-FOXO1 signaling pathway. AB - Recent studies in mice have shown that resveratrol can protect the liver from fat accumulation induced by high fat diet. However, the exact mechanism is largely unknown. To explore the possible mechanism, we investigated the anti-lipogenic effect of resveratrol in vitro model. Oil Red O staining revealed that resveratrol could significantly ameliorate the excessive triglyceride accumulation in HepG2 cells induced by palmitate. The results of RT-PCR and Western blotting showed that resveratrol upregulated the expression of Sirt1 and forkhead box O1 (FOXO1), whereas downregulated the expression of sterol regulatory element binding protein1 (SREBP1). Moreover, resveratrol was shown to inhibit the activity of SREBP1, as evaluated by immunofluorescence assay. Our results suggest that resveratrol may attenuate fat deposition by inhibiting SREBP1 expression via Sirt1-FOXO1 pathway and thus may have application for the treatment of NAFLD. PMID- 19285016 TI - Lithium chloride protects retinal neurocytes from nutrient deprivation by promoting DNA non-homologous end-joining. AB - Lithium chloride is a therapeutic agent for treatment of bipolar affective disorders. Increasing numbers of studies have indicated that lithium has neuroprotective effects. However, the molecular mechanisms underlying the actions of lithium have not been fully elucidated. This study aimed to investigate whether lithium chloride produces neuroprotective function by improving DNA repair pathway in retinal neurocyte. In vitro, the primary cultured retinal neurocytes (85.7% are MAP-2 positive cells) were treated with lithium chloride, then cultured with serum-free media to simulate the nutrient deprived state resulting from ischemic insult. The neurite outgrowth of the cultured cells increased significantly in a dose-dependent manner when exposed to different levels of lithium chloride. Genomic DNA electrophoresis demonstrated greater DNA integrity of retinal neurocytes when treated with lithium chloride as compared to the control. Moreover, mRNA and protein levels of Ligase IV (involved in DNA non homologous end-joining (NHEJ) pathway) in retinal neurocytes increased with lithium chloride. The end joining activity assay was performed to determine the role of lithium on NHEJ in the presence of extract from retinal neurocytes. The rejoining levels in retinal neurocytes treated with lithium were significantly increased as compared to the control. Furthermore, XRCC4, the Ligase IV partner, and the transcriptional factor, CREB and CTCF, were up-regulated in retinal cells after treating with 1.0mM lithium chloride. Therefore, our data suggest that lithium chloride protects the retinal neural cells from nutrient deprivation in vitro, which may be similar to the mechanism of cell death in glaucoma. The improvement in DNA repair pathway involving in Ligase IV might have an important role in lithium neuroprotection. This study provides new insights into the neural protective mechanisms of lithium chloride. PMID- 19285017 TI - Phosphorylation of CREB, a cyclic AMP responsive element binding protein, contributes partially to lysophosphatidic acid-induced fibroblast cell proliferation. AB - Lysophospholipids regulate a wide array of biological processes including cell survival and proliferation. In our previous studies, we found that in addition to SRE, CRE is required for maximal c-fos promoter activation triggered by lysophosphatidic acid (LPA). c-fos is an early indicator of various cells into the cell cycle after mitogenic stimulation. However, role of CREB activation in LPA-stimulated proliferation has not been elucidated yet. Here, we investigate how LPA induces proliferation in Rat-2 fibroblast cell via CREB activation. We found that total cell number and BrdU-positive cells were increased by LPA. Moreover, levels of c-fos mRNA and cyclin D1 protein were increased via LPA induced CREB phosphorylation. Furthermore, LPA-induced Rat-2 cell proliferation was decreased markedly by ERK inhibitor (U0126) and partially by MSK inhibitor (H89). Taken together, these results suggest that CREB activation could partially up-regulate accumulation of cyclin D1 protein level and proliferation of LPA stimulated Rat-2 fibroblast cells. PMID- 19285018 TI - Characterization of function and regulation of miR-24-1 and miR-31. AB - To date, numerous microRNAs (miRNAs) have been discovered. However, the function of these miRNAs is largely unknown. While our knowledge of miRNA post transcriptional processing has greatly expanded in recent years, we have a limited understanding of the regulation and transcription of miRNA genes. In this study, we characterized two BMP-2 upregulated miRNAs, miR-24-1 and miR-31, in mesenchymal stem cells and showed their opposing function in controlling cellular proliferation, and adipogenesis. Furthermore, we are the first to identify and characterize mouse intronic miR-23b~27b~24-1 and intergenic miR-31 genes. Moreover, we found that pri-miR-23b, pri-miR-27b, and pri-miR-24-1 are transcribed independently and their expression profiles are unique when cells are treated with BMP-2, even though they are located closely together. PMID- 19285019 TI - Thalidomide decreases intrahepatic resistance in cirrhotic rats. AB - Increased intrahepatic resistance (IHR) within cirrhotic liver is caused by increased endotoxemia, cytokines tumor necrosis factor-alpha (TNF-alpha), vasoconstrictor thromboxane A(2) (TXA(2)), and disrupted microvasculatures. We evaluated the effects of thalidomide-related inhibition of TNF-alpha upon the hepatic microcirculation of cirrhosis in rats. Portal venous pressure (PVP), hepatic TNF-alpha, expression of thromboxane synthase (TXS), and leukocyte common antigen (LCA) were measured in bile-duct-ligated (BDL) rats receiving 1 month of thalidomide (BDL-thalido rats). Portal perfusion pressure (PPP), IHR, and hepatic TXA(2) production were measured in the isolated liver perfusion system. Intravital microscopy was used to examine hepatic microvascular disruptions. In BDL-thalido rats, PVP, PPP, IHR, hepatic TXA(2) and TNF-alpha, hydroxyproline content, expression of TXS and LCA, and LPS-induced leukocyte recruitment were significantly decreased. Conversely, hepatic microvascular density and perfused sinusoids were significantly increased. Thalidomide decreased PVP and IHR by reducing hepatic TXA(2) and improving hepatic microvascular disruptions in rats with biliary cirrhosis. PMID- 19285020 TI - Cadmium interferes with the degradation of ATF5 via a post-ubiquitination step of the proteasome degradation pathway. AB - ATF5 is a member of the CREB/ATF family of transcription factors. In the current study, using a transient transfection system to express FLAG epitope fusion proteins of ATF5, we have shown that CdCl(2) or NaAsO(3) increases the protein levels of ATF5 in cells, and that cadmium stabilizes the ATF5 protein. Proteasome inhibitors had a similar effect to cadmium on the cellular accumulation of ATF5. Proteasome inhibition led to an increase in ubiquitinated ATF5, while cadmium did not appear to reduce the extent of ATF5 ubiquitination. ATF5 contains a putative nuclear export signal within its N-terminus. We demonstrated that whereas deletion of N-terminal region resulted in a increase of ATF5 levels, this region does not appear to be involved in the ubiquitination of ATF5. These results indicate that ATF5 is targeted for degradation by the ubiquitin-proteasome pathway, and that cadmium slows the rate of ATF5 degradation via a post ubiquitination mechanism. PMID- 19285021 TI - Regional variation in adipogenesis and IGF regulatory proteins in the fetal baboon. AB - Intrauterine growth rate is associated with body distribution in adulthood suggesting differential response of fetal fat depots to nutritional modifications. We hypothesize that there is regional differences in fetal adipogenesis, in part, due to depot-specific regulation of the availability of insulin growth factors. In near-term baboon fetuses (n=3-5), the subcutaneous abdominal vs. omental preadipocytes had (1) more extensive lipid accumulation as assessed by BODIPY (lipid staining) to DAPI (nuclei) absorbance ratios (mean+/ SEM; 0.51+/-0.21, 0.35+/-0.09, p<0.05), (2) lower (p<0.05) secretion of IGF binding protein 4 (9.6+/-1.2 vs. 17.4+/-2.8 ng/ml) and its protease pregnancy associated plasma protein A (24.6+/-1.9 vs. 39.1+/-6.3 microIU/ml), (3) lower protein expression of IGF2 "clearance" receptor in cell lysate (0.28+/-0.03 vs. 0.53+/-0.02 OD U/mm(2), p<0.05); all variables were intermediate in femoral preadipocytes. The regional variation of the adipogenesis and the IGF regulatory pathway set the stage for differential responsiveness of fat depots to external signals. PMID- 19285022 TI - The hepatic circadian clock is preserved in a lipid-induced mouse model of non alcoholic steatohepatitis. AB - Recent studies have correlated metabolic diseases, such as metabolic syndrome and non-alcoholic fatty liver disease, with the circadian clock. However, whether such metabolic changes per se affect the circadian clock remains controversial. To address this, we investigated the daily mRNA expression profiles of clock genes in the liver of a dietary mouse model of non-alcoholic steatohepatitis (NASH) using a custom-made, high-precision DNA chip. C57BL/6J mice fed an atherogenic diet for 5 weeks developed hypercholesterolemia, oxidative stress, and NASH. DNA chip analyses revealed that the atherogenic diet had a great influence on the mRNA expression of a wide range of genes linked to mitochondrial energy production, redox regulation, and carbohydrate and lipid metabolism. However, the rhythmic mRNA expression of the clock genes in the liver remained intact. Most of the circadianly expressed genes also showed 24-h rhythmicity. These findings suggest that the biological clock is protected against such a metabolic derangement as NASH. PMID- 19285023 TI - Related transcriptional enhancer factor-1 induces fibroblast growth factor receptor-1 expression in endothelial cells. AB - Fibroblast growth factor receptor-1 (FGFR-1) has been implicated in the process of cardiogenesis, although the underlying molecular mechanisms are poorly understood. In this study, we report the regulation of FGFR-1 expression by related transcriptional enhancer factor-1 (RTEF-1) in vitro (endothelial cells) and in vivo (RTEF-1 transgenic mice). FGFR-1 promoter activity, FGFR-1 mRNA and protein level were measured in bovine aortic endothelial cells (BAEC) in response to RTEF-1 and in endothelial cells isolated from livers in RTEF-1 transgenic mice. RTEF-1 stimulated FGFR-1 promoter activity in a dose-dependent manner. RTEF 1 enhanced FGFR-1 mRNA (4-fold) and protein expression (3.5-fold) whereas RTEF-1 siRNA decreased FGFR-1 protein expression (4-fold). FGFR-1 mRNA and protein expression were also increased in endothelial cells isolated from livers of RTEF 1 transgenic mice. Furthermore, RTEF-1 enhanced tubule formation whereas this was decreased by RTEF-1 knockdown. Moreover, increased relaxation of microvessels was found in RTEF-1 transgenic mice compared to wild-type mice. Our results indicate that RTEF-1 acts as a transcriptional stimulator of FGFR-1 in endothelial cells through its activation of the FGFR-1 promoter. RTEF-1 thus plays an important role in the regulation of FGFR-1 expression. These findings help further understand FGFR activity in angiogenesis and may lead to new therapeutic targets in ischemic vascular disorders. PMID- 19285024 TI - A novel activity enhances promoter escape of RNA polymerase I. AB - We have characterized a novel transcriptional activity from HeLa cells that is required for ribosomal gene transcription by RNA polymerase I. This activity has a native molecular mass of 16 kDa and does not bind to conventional chromatographic resins. Single-round and immobilized-template experiments revealed that initiation complex formation is independent of the novel activity. Functional studies showed that it stimulates the transition from initiation to elongation, promoter escape. Thus the novel activity does not resemble the mouse initiation/elongation factor TIF-IC but is a true novel entity. PMID- 19285025 TI - AIB1 is required for the acquisition of epithelial growth factor receptor mediated tamoxifen resistance in breast cancer cells. AB - Acquired resistance to tamoxifen has become a serious obstacle in breast cancer treatment. The underlying mechanism responsible for this condition has not been completely elucidated. In this study, a tamoxifen-resistant (Tam-R) MCF-7 breast cancer cell line was developed to mimic the occurrence of acquired tamoxifen resistance as seen in clinical practice. Increased expression levels of HER1, HER2 and the estrogen receptor (ER)-AIB1 complex were found in tamoxifen resistant cells. EGF stimulation and gefitinib inhibition experiments further demonstrated that HER1/HER2 signaling and AIB1 were involved in the proliferation of cells that had acquired Tam resistance. However, when AIB1 was silenced with AIB1-siRNA in Tam-R cells, the cell growth stimulated by the HER1/HER2 signaling pathway was significantly reduced, and the cells were again found to be inhibited by tamoxifen. These results suggest that the AIB1 protein could be a limiting factor in the HER1/HER2-mediated hormone-independent growth of Tam-R cells. Thus, AIB1 may be a new therapeutic target, and the removal of AIB1 may decrease the crosstalk between ER and the HER1/HER2 pathway, resulting in the restoration of tamoxifen sensitivity in tamoxifen-resistant cells. PMID- 19285026 TI - DAZAP1 interacts via its RNA-recognition motifs with the C-termini of other RNA binding proteins. AB - The turnover and translation of many human mRNAs is regulated by AU-rich elements present in their 3?untranslated region, which bind various trans acting factors. We previously identified a trans acting factor that interacts with these cis elements as DAZAP1 (deleted in Azoospermia (DAZ)-Associated Protein 1), whose interaction with the germ cell-specific protein DAZ was disrupted by the phosphorylation of DAZAP1. Here we have identified several other RNA-binding proteins as binding partners for DAZAP1 in non-germinal cells. Unlike DAZ, these interactions occur between the RNA recognition motifs of DAZAP1 and the C-termini of the binding partners and in a phosphorylation-independent manner. The results suggest that DAZAP1 is a component of complexes that are crucial for the degradation and silencing of mRNA. PMID- 19285027 TI - Biotransformation of the neonicotinoid insecticides imidacloprid and thiamethoxam by Pseudomonas sp. 1G. AB - We report the isolation of a Pseudomonas sp. which is able to transform imidacloprid and thiamethoxam under microaerophilic conditions in the presence of an alternate carbon source. This bacterium, Pseudomonas sp. 1G, was isolated from soil with a history of repeated exposure to imidacloprid. Both insecticides were transformed to nitrosoguanidine (NNO), desnitro (NH), and urea (O) metabolites and a transformation pathway is proposed. This is the first conclusive report of bacterial transformation of the 'magic nitro' group which is responsible for the insect selectivity of neonicotinoid insecticides. PMID- 19285028 TI - A functional motif QLYxxYP is essential for osteopontin induced T lymphocyte activation and migration. AB - Osteopontin (OPN) plays an important role in regulating lymphocyte adhesion and cytokine production associated with inflammatory processes and autoimmune diseases. Here we developed and characterized a monoclonal antibody F8E11 specific for human OPN (hOPN). F8E11 could inhibit OPN-induced lymphocyte activation and migration. Epitope mapping showed that F8E11 could specifically recognize the peptide QLYxxYP. In addition, a synthesized mimetic peptide F8P (EEKQLYNKYPDA) could block the binding of F8E11 to hOPN and significantly inhibit the hOPN-induced lymphocyte migration. Moreover, mutations on the QLYxxYP motif of hOPN also markedly diminished its activity for lymphocyte activation and migration. The functioning assay indicated that this novel epitope is critically involved in the lymphocyte migration through activating MAPK/ERK/AP-1 pathway, which can be inhibited by the motif QLYxxYP blocking antibody, F8E11. These results suggest that this novel epitope of OPN may provide a potential therapeutic target for the treatment of T cell mediated-immune diseases. PMID- 19285029 TI - Isolation of functional pure mitochondria by superparamagnetic microbeads. AB - Isolation of mitochondria by current methods relies mainly on their physicochemical properties. Here we describe an alternative approach to obtain functional mitochondria from human cells in a fast, reproducible, and standardized procedure. The new approach is based on superparamagnetic microbeads conjugated to anti-TOM22 antibody. The bead conjugates label the cytoplasmic part of the human mitochondrial membrane protein TOM22 and, thus, allow for a gentle isolation of mitochondria in a high gradient magnetic field. By comparing the MACS (magnetic cell separation) approach with mitochondria isolation methods using differential centrifugation and ultracentrifugation we demonstrate that the MACS approach provides the highest yield of isolated mitochondria. The quality, enrichment, and purity of mitochondria isolated with this protocol are comparable to mitochondria obtained using the ultracentrifuge method, and a typical separation procedure takes only approximately 1 to 2h from initial cell homogenization. Mitochondria isolated with the new approach are sufficient for protein import, blue native gel electrophoresis, and other mitochondrial assays. PMID- 19285030 TI - Characterization of the ATPase activity of a novel chimeric fusion protein consisting of the two nucleotide binding domains of MRP1. AB - Nucleotide Binding Domains (NBDs) are responsible for the ATPase activity of the multidrug resistance protein 1 (MRP1). A series of NBD1-linker-NBD2 chimeric fusion proteins were constructed, expressed and purified, and their ATPase activities were analyzed. We report here that a GST linked NBD1(642-890)-GST NBD2(1286-1531) was able to hydrolyze ATP at a rate of about 4.6 nmol/mg/min (K(m)=2.17 mM, V(max)=12.36 nmol/mg/min), which was comparable to the purified and reconstituted MRP1. In contrast, neither a mixture of NBD1 and GST-NBD2 nor the NBD1-GST-NBD1 fusion protein showed detectable ATPase activity. Additionally, the E1455Q mutant was found to be nonfunctional. Measurements by both MIANS labeling and circular dichroism spectroscopy revealed significant conformational differences in the NBD1-GST-NBD2 chimeric fusion protein compared to the mixture of NBD1 and GST-NBD2. The results suggest a direct interaction mediated by GST between the two NBDs of MRP1 leading to conformational changes which would enhance its ATPase activity. PMID- 19285031 TI - Activation of phospholipase A2 by temporin B: formation of antimicrobial peptide enzyme amyloid-type cofibrils. AB - Phospholipases A2 have been shown to be activated in a concentration dependent manner by a number of antimicrobial peptides, including melittin, magainin 2, indolicidin, and temporins B and L. Here we used fluorescently labelled bee venom PLA2 (PLA2D) and the saturated phospholipid substrate 1,2-dipalmitoyl-glycero-sn 3-phosphocholine (L-DPPC), exhibiting a lag-burst behaviour upon the initiation of the hydrolytic reaction by PLA2. Increasing concentrations of Cys-temporin B and its fluorescent Texas red derivative (TRC-temB) caused progressive shortening of the lag period. TRC-temB/PLA2D interaction was observed by Forster resonance energy transfer (FRET), with maximum efficiency coinciding with the burst in hydrolysis. Subsequently, supramolecular structures became visible by microscopy, revealing amyloid-like fibrils composed of both the activating peptide and PLA2. Reaction products, palmitic acid and 1-palmitoyl-2-lyso-glycero-sn-3 phosphocholine (lysoPC, both at >8 mol%) were required for FRET when using the non-hydrolysable substrate enantiomer 2,3-dipalmitoyl-glycero-sn-1-phosphocholine (D-DPPC). A novel mechanism of PLA2 activation by co-fibril formation and associated conformational changes is suggested. PMID- 19285032 TI - c-ABL tyrosine kinase stabilizes RAD51 chromatin association. AB - The assembly of RAD51 recombinase on DNA substrates at sites of breakage is essential for their repair by homologous recombination repair (HRR). The signaling pathway that triggers RAD51 assembly at damage sites to form subnuclear foci is unclear. Here, we provide evidence that c-ABL, a tyrosine kinase activated by DNA damage which phosphorylates RAD51 on Tyr-315, works at a previously unrecognized, proximal step to initiate RAD51 assembly. We first show that c-ABL associates with chromatin after DNA damage in a manner dependent on its kinase activity. Using RAD51 mutants that are unable to oligomerize to form a nucleoprotein filament, we separate RAD51 assembly on DNA to form foci into two steps: stable chromatin association followed by oligomerization. We show that phosphorylation on Tyr-315 by c-ABL is required for chromatin association of oligomerization-defective RAD51 mutants, but is insufficient to restore oligomerization. Our findings suggest a new model for the regulation of early steps of HRR. PMID- 19285033 TI - Activators of G proteins inhibit GSK-3beta and stabilize beta-Catenin in Xenopus oocytes. AB - Frizzled proteins, the receptors for Wnt ligands have seven hydrophobic transmembrane domains, a structural feature of G protein coupled receptors. Therefore a role for G proteins in the regulation of Wnt signaling has been proposed. Here I have used Xenopus oocytes to study the role of heterotrimeric G proteins in the regulation of GSK-3beta and beta-Catenin, two essential components of the canonical Wnt pathway. In these cells, general activators of G proteins such as GTPgamma-S and AlF4(-) increase beta-Catenin stability and decrease GSK-3beta mediated phosphorylation of the microtubule associated protein, Tau. Among several members of Galpha proteins tested, expression of a constitutively active mutant of Galphaq (GalphaqQL) led to a significant increase in accumulation of beta-Catenin. The stabilization of beta-Catenin mediated by Galphaq was reversed by a Galphaq specific inhibitor, Gp-antagonist 2A, but not by a specific blocking peptide for Galphas. Expression of GalphaqQL also inhibited GSK-3beta-mediated tau phosphorylation in Xenopus oocytes. These results support a role for the Gq class of G proteins in the regulation of Wnt/beta-Catenin signal transduction. PMID- 19285034 TI - MEK1-independent activation of MAPK and MEK1-dependent activation of p70 S6 kinase by stem cell factor (SCF) in ovarian cancer cells. AB - We discovered a stem cell factor (SCF)-triggered, MEK1-independent, and PI3K dependent MAPK activation pathway in the Kit-expressing ovarian cancer cell line HEY. When we knocked down MEK1 with RNA interference (RNAi) to study the function of MEK1 on the proliferation and survival of ovarian cancer cells, we found that impaired cell growth still occurred after MEK1 expression had been suppressed, although MAPK activation remained intact. This suggests that there is MEK1 independent activation of MAPK in the SCF-induced ovarian cancer cell growth process, and that MEK1 still plays a crucial role in maintaining the malignant properties of ovarian cancer cells even when it fails to activate MAPK as expected. PMID- 19285035 TI - Label-free biochemical imaging of heart tissue with high-speed spontaneous Raman microscopy. AB - Label-free imaging is desirable for elucidating morphological and biochemical changes of heart tissue in vivo. Spontaneous Raman microscopy (SRM) provides high chemical contrast without labeling, but presents disadvantage in acquiring images due to low sensitivity and consequent long imaging time. Here, we report a novel technique for label-free imaging of rat heart tissues with high-speed SRM combined with resonance Raman effect of heme proteins. We found that individual cardiomyocytes were identified with resonance Raman signal arising mainly from reduced b- and c-type cytochromes, and that cardiomyocytes and blood vessels were imaged by distinguishing cytochromes from oxy- and deoxy-hemoglobin in intact hearts, while cardiomyocytes and fibrotic tissue were imaged by distinguishing cytochromes from collagen type-I in infarct hearts with principal component analysis. These results suggest the potential of SRM as a label-free high contrast imaging technique, providing a new approach for studying biochemical changes, based on the molecular composition, in the heart. PMID- 19285036 TI - Occurrence of phosphatidyl-D-serine in the rat cerebrum. AB - Phosphatidylserine (PS), a relatively abundant component of mammalian cell membranes, plays important roles in biological processes including apoptosis and cell signaling. It is believed that phosphatidyl-L-serine is the only naturally occurring PS. Here, we describe for the first time the occurrence of phosphatidyl D-serine (D-PS) in rat cerebrum. Quantitative HPLC analysis of the derivatives of serine liberated from PS by hydrolysis revealed that the amount of D-PS was approximately 1% of the total PS in the cerebrum. Enzymatic cleavage of cerebrum PS with phospholipase D and phospholipase C resulted in the release of both isomers of serine and phosphoserine, respectively, providing additional evidence for the existence of D-PS. Free D-serine was incorporated into PS in an in vitro system using a cerebrum extract, and this activity was inhibited by EDTA, suggesting the occurrence of a divalent cation-dependent enzyme that synthesizes D-PS by a base-exchange reaction. PMID- 19285037 TI - Extracellular ATP-induced nuclear Ca2+ transient is mediated by inositol 1,4,5 trisphosphate receptors in mouse pancreatic beta-cells. AB - Extracellular ATP (eATP) induces an intracellular Ca(2+) transient by activating phospholipase C (PLC)-associated P2X4 purinergic receptors, leading to production of inositol 1,4,5-trisphosphate (IP3) and subsequent Ca(2+) release from intracellular stores in mouse pancreatic beta-cells. Using laser scanning confocal microscopy, Ca(2+) indicator fluo-4 AM, and the cell permeable nuclear indicator Hoechst 33342, we examined the properties of eATP-induced Ca(2+) release in pancreatic beta-cell nuclei. eATP induced a higher nuclear Ca(2+) transient in pancreatic beta-cell nuclei than in the cytosol. After pretreatment with thapsigargin (TG), an inhibitor of sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA) pumps, the amplitude of eATP-induced Ca(2+) transients in the nucleus was still much higher than those in the cytosol. This effect of eATP was not altered by inhibition of either the plasma membrane Ca(2+)-ATPase (PMCA) or the plasma membrane Na(+)/Ca(2+) exchanger (NCX) by LaCl(3) or by replacement of Na(+) with N-Methyl-Glucosamine. eATP-induced nuclear Ca(2+) transients were abolished by a cell-permeable IP3R inhibitor, 2-aminoethoxydiphenyl borate (2-APB), but were not blocked by the ryanodine receptor (RyR) antagonist ryanodine. Immunofluorescence studies showed that IP3Rs are expressed on the nuclear envelope of pancreatic beta-cells. These results indicate that eATP triggers nuclear Ca(2+) transients by mobilizing a nuclear Ca(2+) store via nuclear IP3Rs. PMID- 19285038 TI - Secreted cysteine-rich FGF receptor derives from posttranslational processing by furin-like prohormone convertases. AB - Cysteine-rich FGF receptor (CFR) was originally identified as a FGF2 receptor and found to be identical to Golgi complex-localized glycoprotein-1 (GLG1), also known as MG-160, and to a murine E-selectin ligand-1 (ESL-1). Although CFR is a 150-kDa integral membrane glycoprotein that is primarily located in the cis medial Golgi complex, a substantial proportion of CFR is secreted but the underlying mechanism is unknown. CFR contains several possible furin-like proprotein convertase (PC) and matrix metalloproteinase cleavage sites. Cells expressing CFR were treated with the furin protease inhibitor decanoyl-Arg-Val Lys-Arg-chloromethylketone (decCMK) or the MMP-inhibitor GM6001. In the presence of furin-like PC inhibitor, secretion of CFR was almost completely inhibited. Secretion was not affected by the GM6001 inhibitor. The secreted forms were further characterized by creating different mutant CFR proteins with N-terminal and C-terminal tags. Immunoblot analysis and immunofluorescence indicated, that successive endoproteolytical processing of CFR which takes place in the Golgi complex is essential for secretion. Secreted CFR bound to heparan sulphate proteoglycan (HSPG) could trap FGFs and thereby directly competing with tyrosine kinase receptors for FGF binding. PMID- 19285039 TI - Interleukin-6 induces proinflammatory signaling in Schwann cells: a high throughput analysis. AB - Interleukin-6 plays an important role in peripheral nerve regeneration. We recently reported that IL-6 targets Schwann cells in the peripheral nerve for its function. In this study, we analyzed genes whose expression is regulated by IL-6 in a cell line derived from Schwann cells, the peripheral glia, using the Illumina gene microarray. At measurements 3 and 12h after IL-6 treatment, 35 genes were found to be upregulated by IL-6. Most upregulated genes were proinflammatory genes that are known to be induced in inflammatory conditions. Interestingly, the expression of immunoproteasome subunits was upregulated by IL 6 in Schwann cells. Treatment with forskolin, an agent that mimics axonal signaling, suppressed the expression of IL-6-inducible genes. Finally, we found for the first time that sciatic nerve injury induced immunoproteasome expression in vivo. These findings indicate that IL-6 is involved in peripheral nerve regeneration by regulating proinflammatory signaling in Schwann cells. PMID- 19285040 TI - Generation of insulin-secreting cells from adult rat pancreatic ductal epithelial cells induced by hepatocyte growth factor and betacellulin-delta4. AB - The present study was conducted to identify effective factors that could induce differentiation of pancreatic ductal epithelial cells (PDECs) into insulin producing cells. We examined the effect of hepatocyte growth factor (HGF) and betacellulin-delta4 (BTC-delta4) on PDECs. We isolated PDECs from adult rats and stimulated them with HGF and BTC-delta4 for 28days. We observed several characteristics of PDECs after induction, including functional changes in PDECs, differentiation rate, and expression levels of critical transcription factors. We observed that insulin content and glucose-induced insulin secretion were significantly higher in induced PDECs compared to control PDECs. The expression of markers specific to mature beta-cells, i.e., insulin and glucose transporter-2 (GLUT2) was also observed. An analysis of the expression of different transcription factors (PDX-1, Ngn3, NeuroD, and Pax4) revealed that cell-specific gene expression patterns varied from 7 to 28days after treatment. Hence, we conclude that treatment with a combination of HGF and BTC-delta4 is effective in inducing the differentiation of PDECs into insulin-secreting cells. PMID- 19285041 TI - High-fat diet decreases tyrosine hydroxylase mRNA expression irrespective of obesity susceptibility in mice. AB - Tyrosine hydroxylase is the rate-limiting enzyme in the synthesis of dopamine, a key neurotransmitter in the regulation of food intake. This study examined tyrosine hydroxylase mRNA expression in obese mice fed a high-fat diet. After 8 week feeding of high-fat diet mice were classified as diet-induced obese and obese-resistant according to body weight gain. They were then placed on different dietary interventions including a high-fat diet, a low-fat diet and an energy restricted high-fat diet for six weeks. The control group was fed a low-fat diet. The results revealed that tyrosine hydroxylase mRNA expression was significantly decreased in the ventral tegmental area (VTA), ventromedial hypothalamic nucleus (VMH), and substantia nigra (SN) of the high-fat diet-induced obese (-29%, -26% and -26%) and obese-resistant mice (-21%, -24% and -18%) compared to controls. After switching the diet from high to low-fat diet tyrosine hydroxylase mRNA was increased in the VTA, VMH, and SN of the diet-induced obese mice and in the VMH, and SN of the obese-resistant mice. Energy restriction, even with high-fat feeding, reduced tyrosine hydroxylase mRNA expression in the VTA, VMH, and SN compared to controls. In addition, tyrosine hydroxylase mRNA expression in the VTA, VMH, and SN showed a significant negative correlation with plasma leptin levels. This study suggests that the up- or down-regulation of tyrosine hydroxylase mRNA expression in the VTA, VMH, and SN is mainly due to the intake of macronutrient type rather than body weight. PMID- 19285042 TI - Physical fitness, but not acute exercise modulates event-related potential indices for executive control in healthy adolescents. AB - Physical activity and aerobic exercise in particular, promotes health and effective cognitive functioning. To elucidate mechanisms underlying the beneficial effects of physical fitness and acute exercise, behavioral and electrophysiological indices of task preparation and response inhibition as a part of executive functions were assessed in a modified version of an Eriksen flanker task subsequent to an acute bout of aerobic exercise and a period of rest, respectively. 35 higher- and lower-fit adolescents between 13 and 14 years of age participated in a controlled cross-over study design. Results indicate that higher-fit individuals show significantly greater CNV amplitudes, reflecting enhanced task preparation processes, as well as decreased amplitudes in N2, indexing more efficient executive control processes. P3 amplitudes associated with the allocation of attentional and memory control neither showed influences of physical fitness nor the acute bout of exercise. Furthermore, acute aerobic exercise was not related to any of the dependent measures. The current findings suggest that physical fitness, but not an acute bout of aerobic exercise enhances cognitive processing by increasing attentional allocation to stimulus encoding during task preparation. PMID- 19285043 TI - Effect of beta-phenylethylamine on extracellular concentrations of dopamine in the nucleus accumbens and prefrontal cortex. AB - It is known that psychostimulants stimulate dopamine transmission in the nucleus accumbens. In the present study, we examined the effects of systemically administered beta-phenylethylamine (beta-PEA), a psychomotor-stimulating trace amine, on dopamine concentrations in the nucleus accumbens and prefrontal cortex in freely moving rats, using an in vivo microdialysis technique. Intraperitoneal administration of beta-PEA (12.5 and 25 mg/kg) significantly increased extracellular dopamine levels in the nucleus accumbens shell. The observed increase in the dopamine concentration in nucleus accumbens shell dialysate after intraperitoneal administration of 25 mg/kg beta-PEA was inhibited by pre treatment with a dopamine uptake inhibitor, GBR12909 (10 mg/kg, i.p.). In contrast, beta-PEA (25 mg/kg, i.p.) did not affect dopamine release in the nucleus accumbens core. Although a high dose of beta-PEA (50 mg/kg) significantly increased dopamine levels in the nucleus accumbens core, the dopamine increasing effect of beta-PEA was more potent in the nucleus accumbens shell. Systemic administration of 12.5 and 25 mg/kg beta-PEA also increased extracellular dopamine levels in the prefrontal cortex of rats. However, systemic 25 mg/kg beta PEA-induced increases in extracellular dopamine levels were not blocked by GBR12909 within the prefrontal cortex. These results suggest that beta-PEA has a greater effect in the shell than in the core and low-dose beta-PEA stimulates dopamine release in the nucleus accumbens shell through uptake by a dopamine transporter. Similarly, beta-PEA increased extracellular dopamine levels in the prefrontal cortex. Thus, beta-PEA may increase extracellular dopamine concentrations in the mesocorticolimbic pathway. PMID- 19285044 TI - Beta-hydroxybutyrate alters GABA-transaminase activity in cultured astrocytes. AB - The ketogenic diet has long been recognized as an effective treatment for medically refractory epilepsy. Despite nearly a century of use, the mechanisms underlying its clinical efficacy remain unknown. One of the proposed hypotheses for its anti-epileptic actions involves increased GABA concentration in the brain due to ketone bodies that become elevated with a ketogenic diet. In recent years, the notion that astrocytes could play a role in the evolution of abnormal cortical excitability in chronic neurological disorders, such as epilepsy, has received renewed attention. The present study examined the effects of beta hydroxybutyrate, a ketone body, on GABA metabolism in rat primary cultured astrocytes. When beta-hydroxybutyrate was added to culture medium, GABA transaminase (GABA-T) mRNA expression was significantly suppressed in time- and dose-dependent manners. GABA-T enzymatic activity in beta-hydroxybutyrate-treated astrocytes was also suppressed, in accordance with its gene expression. These effects were evident after 3 days of culture, which might coincide with depleted intracellular glycogen. GABA transporter, GAT-1, gene expression was strongly suppressed in cultured astrocytes after 5 days of culture with beta hydroxybutyrate, although other type of GABA transporters did not display significant changes. These results suggest that beta-hydroxybutyrate induced by ketogenic diet may increase GABA concentration in the epileptic brain by suppressing astrocytic GABA degradation, leading to antiepileptic effects. PMID- 19285045 TI - The cortical representation of sensory inputs arising from bone. AB - In the present study, we show that sensory information from bone reaches the discriminative areas of the somatosensory cortices by electrically stimulating the nerve to the cat humerus and recording evoked potentials on the surface of the primary (SI) and secondary (SII) somatosensory cortex. The SI focus was located over the rostral part of the postcruciate cortex, caudal to the lateral aspect of the cruciate sulcus. The SII focus was identified on the anterior ectosylvian gyrus, lateral to the suprasylvian sulcus. These foci were located adjacent to, or within areas that responded to stimulation of the median, ulnar and/or musculocutaneous nerves. The latency (6-11 ms) to onset of cortical responses in SI and SII were indistinguishable (unpaired t-test; P>0.05), and were consistent with activation of A delta fibers in the peripheral nerve. The amplitudes of the cortical responses were graded as a function of stimulus intensity, and may reflect a mechanism for intensity coding. We did not observe long latency cortical responses (50-300 ms) that would be consistent with C fiber activation in the peripheral nerve, and provide evidence that this may be attributable to inhibition of cortical responsiveness following the initial A delta response. Our finding of discrete, short latency evoked potentials (presumably of A delta origin) in the primary and secondary somatosensory cortices, following stimulation of a nerve innervating bone, may reflect a mechanism for the discriminative component of bone pain. PMID- 19285046 TI - Synapse loss regulated by matrix metalloproteinases in traumatic brain injury is associated with hypoxia inducible factor-1alpha expression. AB - The present study assessed the role of matrix metalloproteinase-2 (MMP-2) and -9 in synapse loss after traumatic brain injury (TBI) and the role of hypoxia inducible factor-1alpha (HIF-1alpha), a transcription factor up-regulated during hypoxia, in the regulation of MMP-2 and -9 expression post-TBI. Adult male Sprague-Dawley rats (n=6 per group, 400 g-425 g) were injured using Marmarou's closed-head acceleration impact model and allowed to survive for 1, 4, 24 and 48 h. In another set of experiments, 30 min after TBI, animals were treated with Minocycline (inhibitor of MMPs), or 2-Methoxyestradiol (2ME2, inhibitor of HIF 1alpha) and sacrificed at 4 h after injury. Relative amounts of synaptophysin, a presynaptic vesicular protein, HIF-1alpha, as well as MMP-2 and -9 were assessed by real-time PCR and Western blotting. Activity levels of MMP-2 and -9 were determined by zymography. Synaptophysin expression was significantly (p<0.05) decreased at 1 h through 48 h after TBI. A significant increase in gene and protein expressions of HIF-1alpha, MMP-2 and -9, as well as enzyme activity of MMP-2 and -9 at the same time points was also detected. Inhibition of either MMPs or HIF-1alpha significantly reversed the TBI-induced decrease in synaptophysin. Inhibition of HIF-1alpha reduced expression of MMP-2 and -9. This study showed an early detection of a correlation between synaptic loss and MMP expression after TBI. The data also supports a role for HIF-1alpha in the MMP regulatory cascade in synapse loss after TBI, suggesting potential targets for reducing loss of synaptic terminals. PMID- 19285047 TI - N-(4-Hydroxyphenyl) retinamide potentiated paclitaxel for cell cycle arrest and apoptosis in glioblastoma C6 and RG2 cells. AB - Glioblastoma grows aggressively due to its ability to maintain abnormally high potentials for cell proliferation. The present study examines the synergistic actions of N-(4-hydroxyphenyl) retinamide (4-HPR) and paclitaxel (PTX) to control the growth of rat glioblastoma C6 and RG2 cell lines. 4-HPR induced astrocytic differentiation that was accompanied by increased expression of the tight junction protein e-cadherin and sustained down regulation of Id2 (member of inhibitor of differentiation family), catalytic subunit of rat telomerase reverse transcriptase (rTERT), and proliferating cell nuclear antigen (PCNA). Flow cytometric analysis showed that the microtubule stabilizer PTX caused cell cycle deregulation due to G2/M arrest. This in turn could alter the fate of kinetochore spindle tube dynamics thereby halting cell cycle progression. An interesting observation was the induction of G1/S arrest by a combination of 4-HPR and PTX, altering the G2/M arrest induced by PTX alone. This was further ratified by the upregulation of tumor suppressor protein retinoblastoma, which repressed the expression of the key signaling moieties to induce G1/S arrest. Collectively, the combination of 4-HPR and PTX diminished the survival factors (e.g., rTERT, PCNA, and Bcl-2) to make glioblastoma cells highly prone to apoptosis with activation of cysteine proteases (e.g., calpain, cathepsins, caspase-8, caspase-3). Hence, the combination of 4-HPR and PTX can be considered as an effective therapeutic strategy for controlling the growth of heterogeneous glioblastoma cell populations. PMID- 19285048 TI - Vascular endothelial growth factor-stimulated cerebral microvascular endothelial cells mediate the recruitment of neural stem cells to the neurovascular niche. AB - Endogenous and transplanted neural stem cells (NSC) are highly migratory and display a unique tropism for areas of neuro-pathology. However, signals controlling NSC motility in health and disease are still ill-defined. NSC appear to be intimately associated with the cerebral vasculature and angiogenesis is a hallmark of many neurological disorders. This has led us to investigate the influence of quiescent and angiogenically active human endothelial cells on human NSC migration. In vivo we observed frequent perivascular accumulation of human NSC in the proximity of cerebral microvessels upon induction of angiogenesis by cerebral infusion of vascular endothelial growth factor (VEGF) into the murine brain. We analyzed the in vitro effects of conditioned media from human endothelial cells before and after angiogenic stimulation with VEGF on the migration of human NSC in vitro. Non-stimulated endothelial cells induced a moderate chemotactic migration that was significantly enhanced after angiogenic activation by VEGF. In order to identify cytokines that may function as stimulators of NSC chemotaxis, we screened endothelial cell-conditioned media for the expression of 120 different cytokines. We identified PDGF-BB, RANTES, I-TAC, NAP-2, GROalpha, Ang-2, and M-CSF as endothelial cell-released chemoattractants for human NSC in vitro. VEGF-stimulated cerebral microvascular endothelial cells secreted higher levels of Ang-2 and GROalpha, which in part were responsible for the enhanced chemoattraction of NSC. Our findings support the hypothesis that the angiogenically active microvasculature modulates the local guidance of NSC through endothelial cell-derived chemoattractants. PMID- 19285049 TI - Oxymatrine protects rat brains against permanent focal ischemia and downregulates NF-kappaB expression. AB - BACKGROUND: Oxymatrine is proven to protect ischemic and reperfusion injury in liver, intestine and heart, this effect is via anti-inflammation and anti apoptosis. Whether this protective effect applies to ischemic injury in brain, we therefore investigate the potential neuroprotective role of oxymatrine and the underlying mechanisms. METHODS: Male, Sprague-Dawley rats were randomly assigned to four groups: permanent middle cerebral artery occlusion (pMCAO), high dose (pMCAO+oxymatrine 120 mg/kg), low dose (pMCAO+oxymatrine 60 mg/kg) and sham operated group. We used a permanent middle cerebral artery occlusion model and administered oxymatrine intraperitoneally immediately after cerebral ischemia and once daily on the following days. At 24 h after MCAO, neurological deficit was evaluated using a modified six point scale; brain water content was measured; NF kappaB expression was measured by immunohistochemistry, Western blotting and RT PCR. Infarct volume was analyzed with 2, 3, 5-triphenyltetrazolium chloride (TTC) staining at 72 h. RESULTS: Compared with pMCAO group, neurological deficit in high dose group was improved (P<0.05), infarct volume was decreased (P<0.001) and cerebral edema was alleviated (P<0.05). Consistent with these indices, immunohistochemistry, Western blot and RT-PCR analysis indicated that NF-kappaB expression was significantly decreased in high dose group. Low dose of oxymatrine did not affect NF-kappaB expression in pMCAO rats. CONCLUSIONS: Oxymatrine reduced infarct volume induced by pMCAO, this effect may be through the decreasing of NF-kappaB expression. PMID- 19285050 TI - Post-ischemic hypothermia attenuates loss of the vascular basement membrane proteins, agrin and SPARC, and the blood-brain barrier disruption after global cerebral ischemia. AB - Vascular basement membrane (BM) stabilizes brain vessels and inhibits endothelial cell cycle. Cerebral ischemia causes BM breakdown with the loss of structural BM components including collagens and laminins. In this study, the expression changes of the BM proteoglycan agrin, and the non-structural BM constituent SPARC (BM-40, osteonectin), were studied in brain vessels after global cerebral ischemia. A transient 20-min forebrain ischemia followed by 1, 6 or 24 h of reperfusion was induced in adult Sprague-Dawley rats by combined bilateral common carotid artery occlusion and hypotension (42-45 mm Hg). In a separate group of animals, a mild (32 degrees C) post-ischemic hypothermia was induced for 6 h, starting immediately after ischemia. RNA from approximately 500 brain vessels (20 100 microm) extracted by laser-capture microdissection (LCM) microscopy was used to determine the expression of proteoglycans agrin and SPARC mRNAs by quantitative PCR (Q-PCR). Protein expression was determined by immunohistochemistry in adjacent tissue sections. The BBB permeability was assessed using (3)H-sucrose as an in vivo tracer and by examining fibrinogen immunoreactivity in tissue sections. A transient global brain ischemia resulted in a significant (ANOVA, p<0.05; 6 animals/group) reduction in agrin and SPARC mRNAs in LCM-captured brain vessels 24 h after reperfusion. A time-dependent loss of agrin and SPARC from the BM during reperfusion was also observed by immunochemistry. A 6-h post-ischemic hypothermia reduced SPARC and agrin mRNA and protein losses, BBB transfer constant for (3)H-sucrose as well as fibrinogen extravasation 24 h after reperfusion. It is conluded that a transient post ischemic hypothermia stabilizes brain vessels and reduces BBB disruption in part by preventing proteolytic degradation of regulatory BM constituents, SPARC and agrin. PMID- 19285051 TI - Excitation of cutaneous C nociceptors by intraplantar administration of anandamide. AB - Anandamide has been characterized as both an endocannabinoid and endovanilloid. Consistent with its actions as an endovanilloid, previous studies have demonstrated that anandamide can excite primary sensory neurons in vitro via transient receptor potential vanilloid type one (TRPV1) receptors. In the present study, we sought to determine if anandamide excited cutaneous C nociceptors in vivo and if this excitation correlated with nocifensive behaviors. Using teased fiber electrophysiological methods in the rat, C nociceptors isolated from the tibial nerve with receptive fields (RFs) on the plantar surface of the hindpaw were studied. Injection of anandamide into the RF dose-dependently excited nociceptors at doses of 10 and 100 microg. The TRPV1 receptor antagonists, capsazepine or SB 366791, were applied to the RF to determine if excitation by anandamide was mediated through TRPV1 receptors. Intraplantar injection of either capsazepine (10 microg) or SB 366791 (3 microg) attenuated the excitation produced by 100 microg anandamide. We also determined whether excitation of C nociceptors by anandamide was associated with nocifensive behaviors. Intraplantar injection of 100 microg anandamide produced nocifensive behaviors that were attenuated by pre-treatment with either capsazepine or SB 366791. Furthermore, we determined if intraplantar injection of anandamide altered withdrawal responses to radiant heat. Neither intraplantar injection of anandamide nor vehicle produced antinociception or hyperalgesia to radiant heat. Our results indicate that anandamide excited cutaneous C nociceptors and produced nocifensive behaviors via activation of TRPV1 receptors. PMID- 19285053 TI - Thalamic neuron hyperexcitability and enlarged receptive fields in the STZ model of diabetic pain. AB - Distal limb pain in diabetes mellitus is frequently attributed to hyperexcitability of primary afferents associated with peripheral neuropathy. However, prior studies have demonstrated that, after traumatic nerve injury, hyperexcitability develops not only within primary afferents but also within pain signalling neurons of the spinal cord dorsal horn and thalamic ventral posterolateral (VPL) nucleus, establishing a basis for tiered central pain generators or amplifiers. In this study we asked whether hyperexcitability develops within thalamic neurons in experimental painful diabetes. Diabetes was induced in adult male Sprague-Dawley rats with streptozotocin (STZ). Behavioral testing for tactile allodynia, performed one week prior to STZ injection and weekly thereafter, indicated that, by six weeks after STZ injection, mechanical allodynia had developed (mechanical withdrawal threshold <4 g, STZ; 21.75 g, control). Thalamic unit recordings were obtained from the VPL nucleus at seven weeks after STZ injection, in rats that met a criterion withdrawal threshold of <4 g, at a time when mean glucose level for control rats was 104.8+/-2.9, and for diabetic rats was 420.1+/-42.0. Our analysis shows that, in this model of diabetic neuropathic pain, thalamic VPL neurons develop hyperexcitability, with increased responses to phasic brush, press, and pinch stimuli applied to identified peripheral receptive fields. VPL neurons from diabetic rats also display enhanced spontaneous activity, independent of ascending afferent barrage, and enlarged receptive fields. These results suggest that aberrant levels of spontaneous activity and hyper-responsiveness of VPL thalamic neurons may contribute to diabetic neuropathic pain. PMID- 19285052 TI - Estradiol and lithium chloride specifically alter NMDA receptor subunit NR1 mRNA and excitotoxicity in primary cultures. AB - Glutamate facilitates calcium influx via NMDAR, and excess calcium influx increases excitotoxicity--a pathological characteristic of neurological diseases. Both 17beta-estradiol (E2) and lithium influence NMDAR expression/signaling and excitotoxicity. This led us to hypothesize that combined E2 and lithium will alter NMDAR expression and excitotoxicity. We tested this hypothesis using primary cell cultures from the cortex and hippocampus of C57BL/6J fetal mice pretreated with E2, lithium chloride (LiCl) and combined E2/LiCl for 12, 24 or 48 h. We examined cultures for brain cell type and changes in cell type caused by experimental procedures using glia and neuron gene specific primers. These cultures expressed increased glial fibrillary acidic protein (GFAP) mRNA with low neurofilament-heavy chain (NF-H) mRNA expression. Subsequent analysis of cortical cell cultures indicated that combined E2/LiCl decreased NR1 mRNA expression after a 12 and 48 h treatment period. Combined E2/LiCl also reduced NR1 mRNA expression in hippocampal cultures but only after a 48 h treatment period. LiCl-treated hippocampal cultures also reduced NR1 mRNA expression after a 24 and 48 h treatment. We next examined the response of 48 h pretreated cultures to a toxic level of glutamate. Excitotoxicity was measured using fluorescein diacetate/propidium iodide (FDA/PI) cell viability assay. Results from FDA/PI assay revealed that LiCl pretreatment increased viability for cortical cultures while E2 and combined E2/LiCl reduced viability. All pretreatments for hippocampal cultures failed to increase viability. Our results showed combined E2/LiCl reduced NR1 mRNA and prevented protection against glutamate excitotoxicity in glial primary cultures. PMID- 19285054 TI - Association of ABCB1 gene polymorphisms with plasma lipid and apolipoprotein concentrations in the STANISLAS cohort. AB - BACKGROUND: While involvement of ABCB1 is well known in drug transport, its metabolite transport role is not so well understood. Like other ABC transporters, ABCB1 might be implicated in cholesterol homeostasis and ABCB1 polymorphisms which are responsible for drug resistance might affect lipid homeostasis. Our objective was thus to investigate the implication of ABCB1 polymorphisms and haplotypes in the genetic variability of lipid constituents in healthy people. METHODS: T-129C, G-1A, A61G, G1199A, C1236T, T-76A, G2677T/A and C3435T polymorphisms were genotyped in 371 supposed healthy individuals from the STANISLAS cohort. Each polymorphism was tested with plasma concentrations of total cholesterol, HDL and LDL cholesterol, triglycerides and apolipoproteins A1, B, C3 and E. RESULTS: After adjustment for covariates, carriers of at least one 3435T allele had a significant higher level of apolipoprotein A1 (p = 0.005). In addition, significant correlations were observed in a sex-dependent manner. Women carrying either T-76 or 1236T allele (tendency with G-1 and 2677T/A) had lower total cholesterol (p < or = 0.01) and apolipoprotein B (T-76 exclusively, p=0.002). Haplotypes analysis was not more informative than the single polymorphisms except G2677T/A-C3435T haplotypes for apolipoprotein A1 concentration. CONCLUSION: ABCB1 polymorphisms contribute to the genetic variability of plasma values of lipids and lipoproteins in healthy people. PMID- 19285055 TI - Reference range of thyroid hormones in healthy school-age children: country-wide data from India. AB - OBJECTIVE: This study was planned to obtain normative data of thyroid functions in school-age children from different regions of India. DESIGN AND METHODS: Students from 36 schools involving 13 states across four geographical zones of India were evaluated for goiter. Subjects who consented, underwent evaluation for serum FT3, FT4, TSH, anti-TPO antibodies and thyroid ultrasound. From this, a "reference population" was obtained by excluding those with personal or family history of thyroid disease, use of thyroid medications, goiter, hypoechogenicity or nodularity on ultrasound or positive anti-thyroid antibodies. RESULTS: Of 24,685 students clinically evaluated, 8665 formed part of the study. The reference population comprised 5343 subjects. The mean, median, 3rd and 97th percentiles of FT3, FT4 and TSH for each year (6-17 years) were obtained. CONCLUSIONS: This community based study in Indian school-age children provides reference intervals for thyroid hormones and evidence against narrowing the TSH reference range. PMID- 19285056 TI - Membrane permeability coefficients of murine primary neural brain cells in the presence of cryoprotectant. AB - Neural cells isolated from the brain have a number of research and clinical applications, including transplantation to patients with neurodegenerative conditions. Tissue supply is one of the major limiting factors to clinical transplantation. Cryopreservation of primary neural cells would improve supply, aid in organisation of transplantation surgery and facilitate research. To date, cryopreservation using standard methods has resulted in reduced yield and/or viability of primary neural tissue. In order to optimise freezing protocols specifically for such cells, the non-osmotic volume (V(b)), water permeability (L(p)) and permeability to cryoprotectant (P(cpa)) were determined. Murine foetal brain tissue from the ganglionic eminence (GE), ventral mesencephalon (VM), or neocortical mantle (Ctx) was trypsinised to a single cell suspension. To determine V(b,) cell volume was measured after exposure to anisotonic solutions of sucrose (150-1500 mOsmol/kg). L(p) (mum/min.atm) and P(cpa) (mum/s) were determined for GE cells by measuring cell volume during exposure to 1.5 mol/l cryoprotectant. Cell volume was determined using an electronic particle counting method. V(b) was 27% for Ctx and GE, and 30% for VM. The osmotic response of GE cells was similar in the presence of propane-1,2-diol and dimethyl sulphoxide. In the presence of ethylene glycol, cell volume decrease was greater on initial exposure to cryoprotectant and recovery slower. Differences in L(p,) but not P(cpa), were found between cryoprotectants. The present results provide key parameters for optimisation of freezing protocols for cryopreservation of primary foetal brain tissues for application in neural cell transplantation. PMID- 19285057 TI - Regulation of Drosophila embryonic tracheogenesis by dVHL and hypoxia. AB - The tracheal system of Drosophila melanogaster is an interconnected network of gas-filled epithelial tubes that develops during embryogenesis and functions as the main gas-exchange organ in the larva. Larval tracheal cells respond to hypoxia by activating a program of branching and growth driven by HIF-1alpha/sima dependent expression of the breathless (btl) FGF receptor. By contrast, the ability of the developing embryonic tracheal system to respond to hypoxia and integrate hard-wired branching programs with sima-driven tracheal remodeling is not well understood. Here we show that embryonic tracheal cells utilize the conserved ubiquitin ligase dVHL to control the HIF-1 alpha/sima hypoxia response pathway, and identify two distinct phases of tracheal development with differing hypoxia sensitivities and outcomes: a relatively hypoxia-resistant 'early' phase during which sima activity conflicts with normal branching and stunts migration, and a relatively hypoxia-sensitive 'late' phase during which the tracheal system uses the dVHL/sima/btl pathway to drive increased branching and growth. Mutations in the archipelago (ago) gene, which antagonizes btl transcription, re-sensitize early embryos to hypoxia, indicating that their relative resistance can be reversed by elevating activity of the btl promoter. These findings reveal a second type of tracheal hypoxic response in which Sima activation conflicts with developmental tracheogenesis, and identify the dVHL and ago ubiquitin ligases as key determinants of hypoxia sensitivity in tracheal cells. The identification of an early stage of tracheal development that is vulnerable to hypoxia is an important addition to models of the invertebrate hypoxic response. PMID- 19285058 TI - Tow (Target of Wingless), a novel repressor of the Hedgehog pathway in Drosophila. AB - Hedgehog (Hh) signalling plays a crucial role in the development and patterning of many tissues in both vertebrates and invertebrates. Aberrations in this pathway lead to severe developmental defects and cancer. Hh signal transduction in receiving cells is a well studied phenomenon; however questions still remain concerning the mechanism of repression of the pathway activator Smoothened (Smo) in the absence of Hh. Here we describe a novel repressor of the Hh pathway, Target of Wingless (Tow). Tow represents the Drosophila homolog of a conserved uncharacterised protein family. We show that Tow acts in Hh receiving cells, where its overexpression represses all levels of Hh signalling, and that this repression occurs upstream or at the level of Smo and downstream of the Hh receptor Patched (Ptc). In addition, we find that like Ptc, overexpression of Tow causes an accumulation of lipophorin in the wing disc. We demonstrate that loss of tow enhances different ptc alleles in a similar manner to another pathway repressor, Suppressor of Fused (SuFu), possibly through mediating Ptc dependant lipophorin internalisation. Combined, these results demonstrate that Tow is an important novel regulator of the Hh pathway in the wing imaginal disc, and may shed light on the mechanism of Ptc repression of Smo. PMID- 19285059 TI - The sigma1 receptor interacts with N-alkyl amines and endogenous sphingolipids. AB - The sigma1 receptor is distinguished for its ability to bind various pharmacological agents including drugs of abuse such as cocaine and methamphetamine. Some endogenous ligands have been identified as putative sigma1 receptor regulators. High affinity ligands for the sigma1 receptor contain a nitrogen atom connected to long alkyl chains. We found that long alkyl chain primary amines including endogenous amines belonging to the sphingolipid family such as D-erythro-sphingosine and sphinganine bind with considerable affinity to the sigma1 receptor but not to the sigma2 receptor. The binding of D-erythro sphingosine to the sigma1 receptor appears to be competitive in nature as assessed against the radioligand [3H]-(+)-pentazocine. Interestingly, the well studied sphingolipid mediator sphingosine-1 phosphate did not bind to the sigma1 or the sigma2 receptor. Sphingosine is converted to sphingosine-1 phosphate by a family of sphingosine kinases that regulate the relative levels of these two bioactive lipids in the cell. The selective binding of sphingosine but not sphingosine-1 phosphate to the sigma1 receptor suggests a mechanism for regulation of sigma1 receptor activity by the sphingosine kinase. We have successfully reconstituted this hypothetical model in HEK-293 cells overexpressing both the sigma1 receptor and sphingosine kinase-1. The data presented here strongly supports sphingosine as an endogenous modulator of the sigma1 receptor. PMID- 19285060 TI - Time-dependent vascular actions of cannabidiol in the rat aorta. AB - We have shown that the major active agent of Cannabis sativa, Delta(9) tetrahydrocannabinol, activates peroxisome proliferator-activated receptor gamma [PPARgamma, O'Sullivan, S.E., Tarling, E.J., Bennett, A.J., Kendall, D.A., Randall, M.D., 2005c. Novel time-dependent vascular actions of delta9 tetrahydrocannabinol mediated by peroxisome proliferator-activated receptor gamma. Biochem. Biophys. Res. Commun. 337, 824-831]. The aim of the present study was to investigate whether another pharmacologically active phytocannabinoid, cannabidiol, similarly activates PPARgamma. Functional vascular studies were carried out in rat aortae in vitro by myography. PPARgamma activation was investigated using reporter gene assays, a PPARgamma competition-binding assay and an adipogenesis assay. Cannabidiol caused time-dependent (over 2 h) vasorelaxation of pre-constricted aortae, sensitive to PPARgamma antagonism (GW9662, 1 microM) and super oxide dismutase inhibition. The vascular effects of cannabidiol were not affected by endothelial denudation, nitric oxide synthase inhibition, pertussis toxin, cannabinoid CB1 or cannabinoid CB2 receptor antagonism, or capsaicin pre-treatment. When aortae were contracted with U46619 in a Ca2+-free buffer, vasorelaxation to cannabidiol was substantially reduced. Furthermore, cannabidiol (1-30 microM) inhibited the contractile response to the re-introduction of Ca2+. In a reporter gene assay, cannabidiol increased the transcriptional activity of PPARgamma. Cannabidiol was also found to bind to PPARgamma and stimulate the differentiation of 3T3-L1 fibroblasts into adipocytes, a PPARgamma-mediated response. These results show that cannabidiol binds to and activates PPARgamma, which partially underlies the time-dependent vascular effects of cannabidiol. However, cannabidiol-induced vasorelaxation in the rat isolated aorta appears to be largely due to calcium channel inhibition. PMID- 19285061 TI - SDF-1alpha up-regulates interleukin-6 through CXCR4, PI3K/Akt, ERK, and NF-kappaB dependent pathway in microglia. AB - Stromal cell-derived factor-1 (SDF-1), also known as CXCL12, and its receptor CXC chemokine receptor 4 (CXCR4) express in various kinds of cells in central nervous system. The SDF-1/CXCR4 signaling pathway is regulated by diverse biological effects. SDF-1 is up-regulated in the ischemic penumbra following stroke and has been known to be associated with the homing of bone marrow cells to injury. However, the effect of SDF-1alpha/CXCR4 on cytokine production in microglia is mostly unknown. Here, we demonstrated that SDF-1alpha enhanced IL-6 production in both primary cultured microglia and BV-2 microglia. We further investigated the signaling pathway involved in IL-6 production stimulated by SDF-1alpha in microglia. SDF-1alpha increased IL-6 production in both protein and mRNA levels. These effects were attenuated by ERK, phosphatidylinositol 3-kinase (PI3K), NF kappaB inhibitors, and IkappaB protease inhibitor. Stimulation of microglia with SDF-1alpha also increased Akt and ERK1/2 phosphorylation. In addition, SDF-1alpha treatment also increased IkappaB kinase alpha/beta (IKK alpha/beta) phosphorylation, IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 phosphorylation at Ser(276), translocation of p65 and p50 from cytosol to nucleus and kappaB-luciferase activity. Moreover, SDF-1alpha-mediated increase of kappaB luciferase activity was inhibited by pre-transfection of DN-p85, DN-Akt or DN ERK2. Increase of IKK alpha/beta phosphorylation and binding of p65 and p50 to the NF-kappaB element were both antagonized by PI3K and ERK inhibitors. Our results demonstrate a mechanism linking SDF-1alpha and IL-6, and provide additional support for the notion that SDF-1alpha plays a regulatory role in microglia activation. PMID- 19285062 TI - Pharmacological characterization of (4R)-alkyl glutamate analogues at the ionotropic glutamate receptors--focus on subtypes iGlu(5-7). AB - The kainic acid (kainate, KA) receptors belong to the class of ionotropic glutamate (iGlu) receptors in the central nervous system. Five subtypes have been identified, which have been termed KA(1,2) and iGlu(5-7). In the search for subtype selective ligands, alpha-amino-5-tert-butyl-3-hydroxy-4 isoxazolyl)propionic acid (ATPA), (4R)-methyl Glu (1a), and E-4-neopentylidene Glu (2f) have all previously been reported as selective agonists for the iGlu(5) receptor subtype. In this paper, we present the pharmacological evaluation of a five-compound series of (4R)-alkyl Glu analogs (1b-e,g) which may be envisaged as conformationally released designs of ATPA and 4-alkylidenes 2a-h. Most notable is the pharmacological profile for (4R)-isopentyl Glu (1g) which shows a 10-fold increase in binding affinity for the iGlu(5) receptor subtype (K(i)=20.5 nM) in comparison with its E-4-alkylidene structural isomer 2g. Furthermore, 1g displays high selectivity over other KA receptor subtypes (KA(1,2) and iGlu(6,7)), AMPA-, and NMDA receptors (2050 and >5000 fold, respectively). PMID- 19285063 TI - Clavulanic acid stimulates sexual behaviour in male rats. AB - Sexual behaviour in rats can be used to predict putative effects on human sexual behaviour. Anecdotic reports exist, that the beta-lactamase inhibitor, clavulanic acid exerts sexual stimulating activities in monkeys. To characterize these pro sexual activities, clavulanic acid was tested in three doses and compared to one dose of a sexually inhibitory dose of the selective serotonin reuptake inhibitor, paroxetine, in sexually-experienced male rats, selected for a moderate level of sexual performance in a standard 30-min test with an oestrus female. After acute administration, clavulanic acid had minor sexual stimulating effects at the highest dose in the number of intromissions and in the first ejaculation series. After sub-chronic 7-days treatment, clavulanic acid increased the number of ejaculations at all three doses and reduced the number of intromissions in the 1st series at the highest dose. After chronic 14 days treatment, a similar but stronger pro-sexual profile was observed. The sexual side effects of paroxetine were as expected, including slight sexual inhibitory effects after acute administration, but somewhat stronger overall inhibitory effects after 7 and 14 days pretreatment, particularly notable in the decreasing number of animals contributing to the 2nd ejaculation series, which was even stronger after 14-days treatment. One week after cessation of treatment, the paroxetine group had completely recovered, whereas the highest dose-group of clavulanic acid still showed some pro-sexual effects. This remarkable pro-sexual activity of clavulanic acid cannot readily be explained by its mechanism of action as a beta-lactamase inhibitor but could be due to unexpected central activity of the compound. PMID- 19285064 TI - Involvement of neuropeptide Y Y(1) receptors in the acute, chronic and withdrawal effects of nicotine on feeding and body weight in rats. AB - We investigated the role of neuropeptide Y Y(1) receptors in acute, chronic and withdrawal effects of nicotine with reference to feeding behavior. Rats were administered with nicotine, neuropeptide Y, neuropeptide Y Y(1) receptor agonist [Leu(31),Pro(34)]neuropeptide Y or antagonist BIBP3226 (N(2)-diphenylacetyl)-N [(4-hydroxy-phenyl)-methyl]-D-arginine amide) via i.c.v. route, and food intake was measured at 2 and 6 h post-injection time-points. While acute nicotine or BIBP3226 reduced food intake, increase was observed following neuropeptide Y or [Leu(31),Pro(34)]neuropeptide Y. Nicotine-induced anorexia was antagonized by pre treatment with neuropeptide Y or [Leu(31),Pro(34)]neuropeptide Y, and potentiated by BIBP3226. Furthermore, effects of chronic nicotine (i.p.) and its withdrawal, alone and in combination with BIBP3226 were evaluated with reference to feeding and body weight. Concurrent administration of BIBP3226 with nicotine prevented the development of tolerance to nicotine-induced anorexia, and withdrawal hyperphagia and weight gain. Moreover, acute BIBP3226 attenuated the hyperphagia following nicotine termination. Additionally, immunocytochemical profile of neuropeptide Y in the hypothalamus was studied following differential nicotine treatments. Acute nicotine treatment dramatically reduced neuropeptide Y immunoreactivity in the arcuate and paraventricular nuclei. Chronic nicotine administration decreased neuropeptide Y immunoreactivity in arcuate, but not in paraventricular nucleus. Nicotine withdrawal resulted in significant increase in the neuropeptide Y immunoreactivity in both the nuclei. Neuropeptide Y immunoreactivity in the lateral hypothalamus did not change following any of the treatments. The results suggest that neuropeptide Y in the arcuate and paraventricular nuclei of hypothalamus may be involved in acute, chronic and withdrawal effects of nicotine on the feeding behavior, possibly via neuropeptide Y Y(1) receptors. PMID- 19285065 TI - Is acetylcholine an autocrine/paracrine growth factor via the nicotinic alpha7 receptor subtype in the human colon cancer cell line HT-29? AB - We used immunochemistry to demonstrate expression of acetylcholine's nicotinic alpha7-receptor subtype in human colon cancer cell line HT-29. Moreover, RT-PCR and immunochemistry showed that choline acetyltransferase and acetylcholine esterase, the enzymes responsible for acetylcholine synthesis and degradation, respectively, localise in HT-29 cells. Bromoacetylcholine bromide, an inhibitor of choline acetyltransferase, significantly attenuated basal cell growth. Our findings suggest that acetylcholine might serve as an autocrine/paracrine-or speculatively, even intracrine-signalling molecule in cell line HT-29, thus contributing to carcinogenesis/cancer progression. PMID- 19285066 TI - Resveratrol inhibits proliferation and promotes apoptosis of osteosarcoma cells. AB - The phytoalexin resveratrol has been described to have chemopreventive and chemotherapeutic effects in several tumor models while its effects on osteosarcoma have not been extensively studied. Additionally, resveratrol is a potent activator of the Sirt1/Sir2 (silent information regulator 2) family of NAD dependent deacetylases which plays a role in calorie restriction-mediated tumor suppression. In the present study, we evaluated the effect of resveratrol on growth and apoptosis in four osteosarcoma cell lines (HOS, Saos-2, U-2 OS and MG 63) and a normal human osteoblast cell line (NHOst). We found that Sirt1 protein was relatively higher expressed in the tumor cells than normal osteoblasts. Consistently, resveratrol induced apoptosis in a dose-dependent fashion in the osteosarcoma cells but had minor effect on normal osteoblasts. Also, a similar effect could be elicited by another Sirt1 activator, isonicotinamide. In addition, the pro-apoptotic effect of resveratrol could be enhanced by nutrition restriction elicited by l-asparaginase. We postulate that these effects by resveratrol are mediated via Sirt1 but further studies are needed to confirm or refute this theory. PMID- 19285067 TI - In vitro and in vivo pharmacological characterization of PF-01354082, a novel partial agonist selective for the 5-HT(4) receptor. AB - The pharmacological profile of PF-01354082, a selective 5-HT(4) receptor partial agonist, was investigated. PF-01354082 displayed high affinity for human 5-HT(4d) and dog 5-HT(4h) receptors in binding studies, having Ki values of 2.0 nM and 4.2 nM, respectively. By contrast, PF-01354082 did not show significant affinity for several other 5-HT receptors (5-HT(1A), 5-HT(1B), 5-HT(1D), 5-HT(2A), 5-HT(2B), 5 HT(2C), 5-HT(3A), and 5-HT(7)) or the dopamine D(2long) receptor. Functional assays using either cells expressing human recombinant 5-HT(4d) receptors or rat tunica muscularis mucosae demonstrated that PF-01354082 exhibited partial agonist activity at the 5-HT(4) receptor. The effects of PF-01354082 on in vitro receptor binding, ion channel activity, and sites of uptake were further investigated. PF 01354082 did not show biologically relevant binding activity at concentrations up to 10 microM except for binding to the 5-HT(4e) receptor. Furthermore, PF 01354082 decreased I(HERG) current by only 11% at a concentration of 300 microM, indicating that the compound had greater than 150,000-fold selectivity for the human 5-HT(4d) receptor over hERG channels. An in vivo study using a gastric motility model in conscious dogs demonstrated that oral administration of PF 01354082 resulted in marked and sustained stimulation of gastric motility in a dose-dependent manner. These results indicate that PF-01354082 is an orally active, highly selective, partial agonist of the human 5-HT(4) receptor that is expected to exert a favorable effect on gastrointestinal motor disorders with reduced adverse effects mediated by other related receptors. PMID- 19285068 TI - Role of Ceacam1 in VEGF induced vasculogenesis of murine embryonic stem cell derived embryoid bodies in 3D culture. AB - CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1), a type I transmembrane glycoprotein involved in cell-cell adhesion has been shown to act as an angiogenic factor for mouse and human endothelial cells. Based on the ability of CEACAM1 to initiate lumen formation in human mammary epithelial cells grown in 3D culture (Matrigel), we hypothesized that murine CEACAM1 may play a similar role in vasculogenesis. In order to test this hypothesis, murine embryonic stem (ES) cells stimulated with VEGF were differentiated into embryoid bodies (EB) for 8 days (-8-0 d) and transferred to Matrigel in the presence or absence of anti-CEACAM1 antibody for an additional 12 days (0-12 d). In the absence of anti-CEACAM1 antibody or in the presence of an isotype control antibody, the EB in Matrigel underwent extensive sprouting, generating lengthy vascular structures with well-defined lumina as demonstrated by confocal microscopy, electron microscopy, and immunohistochemical analysis. Both the length and architecture of the vascular tubes were inhibited by anti-CEACAM1 mAb CC1, a mAb that blocks the cell-cell adhesion functions of CEACAM1, thus demonstrating a critical role for this cell-cell adhesion molecule in generating and maintaining vasculogenesis. QRT-PCR analysis of the VEGF treated ES cells grown under conditions that convert them to EB revealed expression of Ceacam1 as early as -5 to -3 d reaching a maximum at day 0 at which time EBs were transferred to Matrigel, thereafter levels at first declined and then increased over time. Other markers of vasculogenesis including Pecam1, VE-Cad, and Tie-1 were not detected until day 0 when EBs were transferred to Matrigel followed by a steady increase in levels, indicating later roles in vasculogenesis. In contrast, Tie-2 and Flk-1 (VEGFR2) were detected on day five of EB formation reaching a maximum at day 0 on transfer to Matrigel, similar to Ceacam1, but after which Tie 2 declined over time, while Flk-1 increased over time. QRT-PCR analysis of the anti-CEACAM1 treated ES cells revealed a significant decrease in the expression of Ceacam1, Pecam1, Tie-1, and Flk-1, while VE-Cad and Tie-2 expression were unaffected. These results suggest that the expression and signaling of CEACAM1 may affect the expression of other factors known to play critical roles in vasculogenesis. Furthermore this 3D model of vasculogenesis in an environment of extracellular matrix may be a useful model for comparison to existing models of angiogenesis. PMID- 19285069 TI - Additional parameters for the morphometry of angiogenesis and lymphangiogenesis in corneal flat mounts. PMID- 19285071 TI - Edaravone in ALS. PMID- 19285070 TI - Dynamics of the expression of intermediate filaments vimentin and desmin during myofibroblast differentiation after corneal injury. AB - Previous studies have suggested that abnormal corneal wound healing in patients after photorefractive keratectomy (PRK) is associated with the appearance of myofibroblasts in the stroma between two and four weeks after surgery. The purpose of this study was to examine potential myofibroblast progenitor cells that might express other filament markers prior to completion of the differentiation pathway that yields alpha-smooth muscle actin (SMA)-expressing myofibroblasts associated with haze localized beneath the epithelial basement membrane after PRK. Twenty-four female rabbits that had -9 diopter PRK were sacrificed at 1 week, 2 weeks, 3 weeks or 4 weeks after surgery. Corneal rims were collected, frozen at -80 degrees C, and analyzed by immunocytochemistry using anti-vimentin, anti-desmin, and anti-SMA antibodies. Double immunostaining was performed for the co-localization of SMA with vimentin or desmin with SMA. An increase in vimentin expression in stromal cells is noted as early as 1 week after PRK in the rabbit cornea. As the healing response continues at two or three weeks after surgery, many stromal cells expressing vimentin also begin to express desmin and SMA. By 4 weeks after the surgery most, if not all, myofibroblasts express vimentin, desmin and SMA. Generalized least squares regression analysis showed that there was strong evidence that each of the marker groups differed in expression over time compared to the other two (p<0.01). Intermediate filaments- vimentin and desmin co-exist in myofibroblasts along with SMA and may play an important role in corneal remodeling after photorefractive keratectomy. The earliest precursors of myofibroblasts destined to express SMA and desmin are detectible by staining for vimentin at 1 week after surgery. PMID- 19285072 TI - Investigation of the neural correlates underlying action observation in multiple sclerosis patients. AB - Recent fMRI evidence indicates that both the execution and the observation of hand actions in multiple sclerosis (MS) patients increase recruitment of a portion of the so-called mirror neuron system. However, it remains unclear whether this is the expression of a compensatory mechanism for the coding of observed action or whether such a mechanism represents a rather unspecific functional adaptation process. Here we used fMRI on early relapsing remitting MS (RRMS) patients to clarify this issue. Functional images of 15 right-handed early RRMS patients and of 15 sex- and age-matched right-handed healthy controls were acquired using a 1.5 T scanner. During scanning, participants simply observed images depicting a human hand either grasping an object or resting alongside an object. As shown by a between-group analysis, when compared to controls, RRMS patients revealed a robust increase of activation in an extensive network of brain regions including frontal, parietal, temporal and visual areas usually activated during action observation. However, this pattern of hemodynamic activity was completely independent of the type of observed hand-object interaction as revealed by the lack of any significant between-group interaction. Our findings are in line with previous fMRI evidence demonstrating cortical reorganization in MS patients during action observation. However, based on our findings we go one step further and suggest that such functional cortical changes may be the expression of a generalized and unspecific compensatory mechanism, that is not necessarily involved in action understanding. PMID- 19285073 TI - A soluble activin type IIB receptor improves function in a mouse model of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a neurologic disease characterized by progressive weakness that results in death within a few years of onset by respiratory failure. Myostatin is a member of the TGF-beta superfamily that is predominantly expressed in muscle and acts as a negative regulator of muscle growth. Attenuating myostatin has previously been shown to produce increased muscle mass and strength in normal and disease animal models. In this study, a mouse model of ALS (SOD1(G93A) transgenic mice) was treated with a soluble activin receptor, type IIB (ActRIIB.mFc) which is a putative endogenous signaling receptor for myostatin in addition to other ligands of the TGF-beta superfamily. ActRIIB.mFc treatment produces a delay in the onset of weakness, an increase in body weight and grip strength, and an enlargement of muscle size whether initiated pre-symptomatically or after symptom onset. Treatment with ActRIIB.mFc did not increase survival or neuromuscular junction innervation in SOD1(G93A) transgenic mice. Pharmacologic treatment with ActRIIB.mFc was superior in all measurements to genetic deletion of myostatin in SOD1(G93A) transgenic mice. The improved function of SOD1(G93A) transgenic mice following treatment with ActRIIB.mFc is encouraging for the development of TGF-beta pathway inhibitors to increase muscle strength in patients with ALS. PMID- 19285074 TI - Trypanosoma cruzi: in vitro activity of Epoxy-alpha-Lap, a derivative of alpha lapachone, on trypomastigote and amastigote forms. AB - Chagas disease is an endemic parasitic infection caused by Trypanosomacruzi that affects 18-20 million people in Central and South America. Recently we described the Epoxy-alpha-Lap, an oxyran derivative of alpha-lapachone, which presents a low toxicity profile and a high inhibitory activity against T.cruzi epimastigotes forms, the non-infective form of this parasite. In this work we described the trypanocidal effects of Epoxy-alpha-Lap on extracellular (trypomastigote) and intracellular (amastigote) infective forms of two T. cruzi strains (Y and Colombian) known by their different infective profile. Our results showed that Epoxy-alpha-Lap is lethal to trypomastigote Y and Colombian strains (97% and 84%, respectively). Interestingly, Epoxy-alpha-Lap also showed a trypanocidal effect in human macrophage infected with T. cruzi Y (85.6%) and Colombian (71.9%) strains amastigote forms. Similar effects were observed on T. cruzi amastigote infected Vero cells (96.4% and 95.0%, respectively). Our results pointed Epoxy alpha-Lap as a potential candidate for Chagas disease chemotherapy since it presents trypanocidal activity on all T. cruzi forms with low) toxicity profile. PMID- 19285075 TI - Is Giardia a significant pathogen in production animals? AB - Although Giardia duodenalis is recognised worldwide as the most important parasitic cause of gastro-intestinal disorder in human patients, the relevance of infection in production animals is prone to debate. Since the 1980s, clinical disease has been associated with giardiasis in production animals, both in natural conditions and in experimental studies. However, most Giardia research is focussed on the relevance of production animals as a reservoir for zoonotic transmission. In this study, the current knowledge on clinical relevance of giardiasis in production animals is reviewed, along with the diagnosis, treatment and control of infection. Furthermore, future research objectives are discussed. PMID- 19285076 TI - X-ray structure of a two-domain type laccase: a missing link in the evolution of multi-copper proteins. AB - A multi-copper protein with two cupredoxin-like domains was identified from our in-house metagenomic database. The recombinant protein, mgLAC, contained four copper ions/subunits, oxidized various phenolic and non-phenolic substrates, and had spectroscopic properties similar to common laccases. X-ray structure analysis revealed a homotrimeric architecture for this enzyme, which resembles nitrite reductase (NIR). However, a difference in copper coordination was found at the domain interface. mgLAC contains a T2/T3 tri-nuclear copper cluster at this site, whereas a mononuclear T2 copper occupies this position in NIR. The trimer is thus an essential part of the architecture of two-domain multi-copper proteins, and mgLAC may be an evolutionary precursor of NIR. PMID- 19285077 TI - SIRT1 regulates tyrosine hydroxylase expression and differentiation of neuroblastoma cells via FOXO3a. AB - To examine the function of SIRT1 in neuronal differentiation, we employed all trans retinoic acid (ATRA)-induced differentiation of neuroblastoma cells. Nicotinamide inhibited neurite outgrowth and tyrosine hydroxylase (TH) expression. Inhibition of PARP or histone deacetylase did not inhibit TH expression, showing the effect to be SIRT1 specific. Expression of FOXO3a and its target proteins were increased during the differentiation and reduced by nicotinamide. FOXO3a deacetylation was increased by ATRA and blocked by nicotinamide. SIRT1 and FOXO3a siRNA inhibited ATRA-induced up-regulation of TH and differentiation. Taken together, these results indicate that SIRT1 is involved in ATRA-induced differentiation of neuroblastoma cells via FOXO3a. PMID- 19285078 TI - Plant Rho-type (Rop) GTPase-dependent activation of receptor-like cytoplasmic kinases in vitro. AB - Plants have evolved distinct mechanisms to link Rho-type (Rop) GTPases to downstream signaling pathways as compared to other eukaryotes. Here, experimental data are provided that members of the Medicago, as well as Arabidopsis, receptor like cytoplasmic kinase family (RLCK Class VI) were strongly and specifically activated by GTP-bound Rop GTPases in vitro. Deletion analysis indicated that the residues implicated in the interaction might be distributed on various parts of the kinases. Using a chimaeric Rop GTPase protein, the importance of the Rho insert region in kinase activation could also be verified. These data strengthen the possibility that RLCKs may serve as Rop GTPase effectors in planta. PMID- 19285079 TI - Lyn inhibits osteoclast differentiation by interfering with PLCgamma1-mediated Ca2+ signaling. AB - Osteoclasts differentiate from macrophage-lineage cells to become specialized for bone resorption function. By a proteomics approach, we found that Lyn was down regulated by the osteoclast differentiation factor, receptor activator of NF kappaB ligand (RANKL). The forced reduction of Lyn caused a striking increase in the RANKL-induced PLCgamma1, Ca(2+), and NFATc1 responses during differentiation. These data suggest that Lyn plays a negative role in osteoclastogenesis by interfering with the PLCgamma1-mediated Ca(2+) signaling that leads to NFATc1 activation. Consistent with the in vitro results, in vivo injection of Lyn specific siRNA into mice calvariae provoked a fulminant bone resorption. Our study provides the first evidence of the involvement of Lyn in the negative regulation of osteoclastogenesis by RANKL. PMID- 19285080 TI - Crystal structure of a soluble decoy receptor IL-22BP bound to interleukin-22. AB - Interleukin-22 (IL-22) plays an important role in the regulation of immune and inflammatory responses in mammals. The IL-22 binding protein (IL-22BP), a soluble receptor that specifically binds IL-22, prevents the IL-22/interleukin-22 receptor 1 (IL-22R1)/interleukin-10 receptor 2 (IL-10R2) complex assembly and blocks IL-22 biological activity. Here we present the crystal structure of the IL 22/IL-22BP complex at 2.75 A resolution. The structure reveals IL-22BP residues critical for IL-22 binding, which were confirmed by site-directed mutagenesis and functional studies. Comparison of IL-22/IL-22BP and IL-22/IL-22R1 crystal structures shows that both receptors display an overlapping IL-22 binding surface, which is consistent with the inhibitory role played by IL-22 binding protein. PMID- 19285081 TI - Automation of the ELISpot assay for high-throughput detection of antigen-specific T-cell responses. AB - The enzyme linked immunospot (ELISpot) assay is a fundamental tool in cellular immunology, providing both quantitative and qualitative information on cellular cytokine responses to defined antigens. It enables the comprehensive screening of patient derived peripheral blood mononuclear cells to reveal the antigenic restriction of T-cell responses and is an emerging technique in clinical laboratory investigation of certain infectious diseases. As with all cellular based assays, the final results of the assay are dependent on a number of technical variables that may impact precision if not highly standardised between operators. When studies that are large scale or using multiple antigens are set up manually, these assays may be labour intensive, have many manual handling steps, are subject to data and sample integrity failure and may show large inter operator variability. Here we describe the successful automated performance of the interferon (IFN)-gamma ELISpot assay from cell counting through to electronic capture of cytokine quantitation and present the results of a comparison between automated and manual performance of the ELISpot assay. The mean number of spot forming units enumerated by both methods for limiting dilutions of CMV, EBV and influenza (CEF)-derived peptides in six healthy individuals were highly correlated (r>0.83, p<0.05). The precision results from the automated system compared favourably with the manual ELISpot and further ensured electronic tracking, increased through-put and reduced turnaround time. PMID- 19285082 TI - Cardioprotection by metabolic shut-down and gradual wake-up. AB - Mitochondria play a critical role in cardiac function, and are also increasingly recognized as end effectors for various cardioprotective signaling pathways. Mitochondria use oxygen as a substrate, so by default their respiration is inhibited during hypoxia/ischemia. However, at reperfusion a surge of oxygen and metabolic substrates into the cell is thought to lead to rapid reestablishment of respiration, a burst of reactive oxygen species (ROS) generation and mitochondrial Ca(2+) overload. Subsequently these events precipitate opening of the mitochondrial permeability transition (PT) pore, which leads to myocardial cell death and dysfunction. Given that mitochondrial respiration is already inhibited during hypoxia/ischemia, it is somewhat surprising that many respiratory inhibitors can improve recovery from ischemia-reperfusion (IR) injury. In addition ischemic preconditioning (IPC), in which short non-lethal cycles of IR can protect against subsequent prolonged IR injury, is known to lead to endogenous inhibition of several respiratory complexes and glycolysis. This has led to a hypothesis that the wash-out of inhibitors or reversal of endogenous inhibition at reperfusion may afford protection by facilitating a more gradual wake-up of mitochondrial function, thereby avoiding a burst of ROS and Ca(2+) overload. This paper will review the evidence in support of this hypothesis, with a focus on inhibition of each of the mitochondrial respiratory complexes. PMID- 19285083 TI - Action potential clamp and chloroquine sensitivity of mutant Kir2.1 channels responsible for variant 3 short QT syndrome. AB - Recently identified genetic forms of short QT syndrome (SQTS) are associated with an increased risk of arrhythmia and sudden death. The SQT3 variant is associated with an amino-acid substitution (D172N) in the KCNJ2-encoded Kir2.1 K+ channel. In this study, whole-cell action potential (AP) clamp recording from transiently transfected Chinese Hamster Ovary cells at 37 degrees C showed marked augmentation of outward Kir2.1 current through D172N channels, associated with right-ward voltage-shifts of peak repolarizing current during both ventricular and atrial AP commands. Peak outward current elicited by ventricular AP commands was inhibited by chloroquine with an IC50 of 2.45 microM for wild-type (WT) Kir2.1, of 3.30 microM for D172N-Kir2.1 alone and of 3.11 microM for co-expressed WT and D172N (P>0.05 for all). These findings establish chloroquine as an effective inhibitor of SQT3 mutant Kir2.1 channels. PMID- 19285084 TI - Crystal structures of the histo-aspartic protease (HAP) from Plasmodium falciparum. AB - The structures of recombinant histo-aspartic protease (HAP) from malaria-causing parasite Plasmodium falciparum as apoenzyme and in complex with two inhibitors, pepstatin A and KNI-10006, were solved at 2.5-, 3.3-, and 3.05-A resolutions, respectively. In the apoenzyme crystals, HAP forms a tight dimer not seen previously in any aspartic protease. The interactions between the monomers affect the conformation of two flexible loops, the functionally important "flap" (residues 70-83) and its structural equivalent in the C-terminal domain (residues 238-245), as well as the orientation of helix 225-235. The flap is found in an open conformation in the apoenzyme. Unexpectedly, the active site of the apoenzyme contains a zinc ion tightly bound to His32 and Asp215 from one monomer and to Glu278A from the other monomer, with the coordination of Zn resembling that seen in metalloproteases. The flap is closed in the structure of the pepstatin A complex, whereas it is open in the complex with KNI-10006. Although the binding mode of pepstatin A is significantly different from that in other pepsin-like aspartic proteases, its location in the active site makes unlikely the previously proposed hypothesis that HAP is a serine protease. The binding mode of KNI-10006 is unusual compared with the binding of other inhibitors from the KNI series to aspartic proteases. The novel features of the HAP active site could facilitate design of specific inhibitors used in the development of antimalarial drugs. PMID- 19285086 TI - 9-Angstrom structure of a microtubule-bound mitotic motor. AB - Kinesin-5 (K5) motors are important components of the microtubule (MT)-based cell division machinery and are targets for small-molecule inhibitors currently in cancer clinical trials. However, the nature of the K5-MT interaction and the regulatory mechanisms that control it remain unclear. Using cryo-electron microscopy and image processing, we calculated the structure of a K5 motor bound to MTs at 9 A resolution, providing insight into this important interaction. Our reconstruction reveals the K5 motor domain in an ATP-like conformation in which MT binding induces the conserved nucleotide-sensing switch I and II loops to form a compact subdomain around the bound nucleotide. Our reconstruction also reveals a novel conformation for the K5-specific drug-binding loop 5, suggesting a possible role for it in switching K5s between force generation and diffusional modes of MT binding. Our data thus shed light on regulation of the interaction between spindle components important for chromosome segregation. PMID- 19285085 TI - The genome of Bacillus subtilis bacteriophage SPO1. AB - We report the genome sequence of Bacillus subtilis phage SPO1. The unique genome sequence is 132,562 bp long, and DNA packaged in the virion (the chromosome) has a 13,185-bp terminal redundancy, giving a total of 145,747 bp. We predict 204 protein-coding genes and 5 tRNA genes, and we correlate these findings with the extensive body of investigations of SPO1, including studies of the functions of the 61 previously defined genes and studies of the virion structure. Sixty-nine percent of the encoded proteins show no similarity to any previously known protein. We identify 107 probable transcription promoters; most are members of the promoter classes identified in earlier studies, but we also see a new class that has the same sequence as the host sigma K promoters. We find three genes encoding potential new transcription factors, one of which is a distant homologue of the host sigma factor K. We also identify 75 probable transcription terminator structures. Promoters and terminators are generally located between genes and together with earlier data give what appears to be a rather complete picture of how phage transcription is regulated. There are complete genome sequences available for five additional phages of Gram-positive hosts that are similar to SPO1 in genome size and in composition and organization of genes. Comparative analysis of SPO1 in the context of these other phages yields insights about SPO1 and the other phages that would not be apparent from the analysis of any one phage alone. These include assigning identities as well as probable functions for several specific genes and inferring evolutionary events in the phages' histories. The comparative analysis also allows us to put SPO1 into a phylogenetic context. We see a pattern similar to what has been noted in phage T4 and its relatives, in which there is minimal successful horizontal exchange of genes among a "core" set of genes that includes most of the virion structural genes and some genes of DNA metabolism, but there is extensive horizontal transfer of genes over the remainder of the genome. There is a correlation between genes in rapid evolutionary flux through these genomes and genes that are small. PMID- 19285087 TI - The 2.1-A crystal structure of native neuroserpin reveals unique structural elements that contribute to conformational instability. AB - Neuroserpin is a selective inhibitor of tissue-type plasminogen activator (tPA) that plays an important role in neuronal plasticity, memory, and learning. We report here the crystal structure of native human neuroserpin at 2.1 A resolution. The structure has a helical reactive center loop and an omega loop between strands 1B and 2B. The omega loop contributes to the inhibition of tPA, as deletion of this motif reduced the association rate constant with tPA by threefold but had no effect on the kinetics of interaction with urokinase. Point mutations in neuroserpin cause the formation of ordered intracellular polymers that underlie dementia familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type neuroserpin is also unstable and readily forms polymers under near-physiological conditions in vitro. This is, in part, due to the substitution of a conserved alanine for serine at position 340. The replacement of Ser340 by Ala increased the melting temperature by 3 degrees C and reduced polymerization as compared to wild-type neuroserpin. Similarly, neuroserpin has Asn-Leu-Val at the end of helix F and thus differs markedly from the Gly-X-Ile consensus sequence of the serpins. Restoration of these amino acids to the consensus sequence increased thermal stability and reduced the polymerization of neuroserpin and its transition to the latent conformer. Moreover, introduction of the consensus sequence into S49P neuroserpin that causes FENIB increased the stability and inhibitory activity of the mutant, as well as blocked polymerization and increased the yield of protein during refolding. These data provide a molecular explanation for the inherent instability of neuroserpin and the effect of point mutations that underlie the dementia FENIB. PMID- 19285088 TI - Bi- or multifunctional opioid peptide drugs. AB - Strategies for the design of bi- or multifunctional drugs are reviewed. A distinction is made between bifunctional drugs interacting in a monovalent fashion with two targets and ligands containing two distinct pharmacophores binding in a bivalent mode to the two binding sites in a receptor heterodimer. Arguments are presented to indicate that some of the so-called "bivalent" ligands reported in the literature are unlikely to simultaneously interact with two binding sites. Aspects related to the development of bi- or multifunctional drugs are illustrated with examples from the field of opioid analgesics. The drug-like properties of the tetrapeptide Dmt(1)[DALDA] with triple action as a micro opioid agonist, norepinephrine uptake inhibitor and releaser of endogenous opioid peptides to produce potent spinal analgesia are reviewed. Rationales for the development of opioid peptides with mixed agonist/antagonist profiles as analgesics with reduced side effects are presented. Progress in the development of mixed micro opioid agonist/delta opioid antagonists with low propensity to produce tolerance and physical dependence is reviewed. Efforts to develop bifunctional peptides containing a micro opioid agonist and a cholecystokinin antagonist or an NK1 receptor antagonist as analgesics expected to produce less tolerance and dependence are also reviewed. A strategy to improve the drug-like properties of bifunctional opioid peptide analgesics is presented. PMID- 19285089 TI - Measurement depth and volume in laser Doppler flowmetry. AB - A new method for estimating the measurement depth and volume in laser Doppler flowmetry (LDF) is presented. The method is based on Monte Carlo simulations of light propagation in tissue. The contribution from each individual Doppler shift is calculated and thereby multiple Doppler shifts are handled correctly. Different LDF setups for both probe based (0.0, 0.25, 0.5, and 1.2 mm source detector separation) and imaging systems (0.5 and 2.0 mm beam diameter) are considered, at the wavelengths 543 nm, 633 nm, and 780 nm. Non-linear speckle pattern effects are accounted for in the imaging system setups. The effects of tissue optical properties, blood concentration, and blood oxygen saturation are evaluated using both homogeneous tissue models and a layered skin model. The results show that the effect on the measurement depth of changing tissue properties is comparable to the effect of changing the system setup, e.g. source detector separation and wavelength. Skin pigmentation was found to have a negligible effect on the measurement depth. Examples of measurement depths are (values are given for a probe based system with 0.25 mm source-detector separation and an imaging system with a 0.5 mm beam diameter, respectively, both operating at 780 nm): muscle - 0.55/0.79 mm; liver - 0.40/0.53 mm; gray matter - 0.48/0.68 mm; white matter - 0.20/0.20 mm; index finger pulp - 0.41/0.53 mm; forearm skin - 0.53/0.56 mm; heat provoked forearm skin - 0.66/0.67 mm. PMID- 19285090 TI - Hemorrhagic shock and nitric oxide release from erythrocytic nitric oxide synthase: a quantitative analysis. AB - A large loss of blood during hemorrhage can result in profound shock, a state of hypotension associated with hemodynamic abnormalities. One of the hypotheses to account for this collapse of homeostasis is that the production of nitric oxide (NO), a gas molecule that dilates blood vessels, is significantly impaired during hemorrhage, resulting in a mismatch between O(2) delivery and the metabolic activity in the tissues. NO can be released from multiple sources in the vasculature. Recent studies have shown that erythrocytes express functional endothelial nitric oxide synthase (NOS3), which potentially serves as an intraluminal NO source. NO delivery from this source is complex: erythrocytes are not only NO producers but also act as potent sinks because of the high affinity of NO for hemoglobin. To test our hypothesis that the loss of erythrocytic NOS3 during hemorrhage contributes to NO deficiency-related shock, we have constructed a multicellular computational model that simulates NO production and transport to allow us to quantify the loss of NO under different hemorrhagic conditions. Our model shows that: (1) during mild hemorrhage and subsequent hemodilution (hematocrit >30%), NO from this intraluminal source is only slightly decreased in the vascular smooth muscle, but the NO level is significantly reduced under severe hemorrhagic conditions (hematocrit <30%); (2) whether a significant amount of NO from this source can be delivered to vascular smooth muscle is strongly dependent on the existence of a protective mechanism for NO delivery; (3) if the expression level of NOS3 on erythrocytes is similar to that on endothelial cells, we estimate approximately 13 pM NO at the vascular smooth muscle from this source when such a protective mechanism is involved. This study provides a basis for detailed studies to characterize the impairment of NO release pathways during hemorrhage and yield important insights for the development of resuscitation methods. PMID- 19285091 TI - Extreme reduction of the capillary lumen in segments of the venular legs of human cutaneous capillaries. AB - While structure and function of precapillary sphincter cells were assured in skin capillaries it is unclear whether segmental reduction of capillary lumina can occur in human capillaries. It has been shown that endothelial cells are able to exert dynamical reactions. Since the first description of the vascular endothelium a great variety of findings were described concerning the active role of capillary endothelial cells in regulation of the capillary lumen applying intravital microscopy. The intravital microscopy was performed in the framework of an observational study to document the long-term stability of capillaries in healthy subjects over many years. In the second year one of the participants showed remarkable changes in capillaries compared to recent recordings. Control recordings were performed 1, 3, 4, 5 and 20 h after the initial examination - until a complete normalization of the capillaries occurred. This case report is documenting for the first time clearly that extreme luminal narrowing of long segments of cutaneous capillaries can also appear in humans, in this case restricted exclusively to the venular leg of the capillaries. Different from the reductions of the capillary lumen induced by electrical irritation in frogs which lasted only for seconds, the capillary lumen narrowing in this case lasted considerably longer, almost over a whole day. It is important to note that the demonstrated findings did not occur in all capillaries and it remains unclear whether such findings are restricted to skin capillaries or might occur also in other regions of the body or even systemically. It could be demonstrated clearly, however, that segmental narrowing of capillary lumina can occur in humans possibly leading to a temporary stillstand of perfusion. PMID- 19285092 TI - Neuronal nicotinic acetylcholine receptors are modulated by zinc. AB - It is known that zinc modulates nicotinic acetylcholine receptors (nAChRs). Here, we studied the effects of zinc on neuronal alpha4beta4 nAChRs, expressed in Xenopus oocytes and activated by nicotine. Membrane ion currents elicited by nicotine (10 nM to 100 microM) were enhanced by zinc (100 microM). Maximal zinc potentiation of the nicotine-activated current (2530%) occurred at 50 nM nicotine, and potentiation gradually decreased as the nicotine concentration increased. The EC(50) and IC(50) for the nicotine-activated current were 639 nM and 14.7 microM nicotine, respectively. Both parameters decreased in the presence of zinc to 160 nM and 4.6 microM, respectively, probably due to an increase of sensitivity of nAChRs for nicotine. We used different concentrations and durations of exposure to nicotine, due to desensitization of nAChRs directly depends on both these factors. With 500 nM nicotine and 20 min washing periods between nicotine applications, zinc potentiation remained constant, 901% for 2 min and 813% for 20 min of nicotine exposure. With continuous application of nicotine, zinc potentiation decreased as the time of nicotine exposure increased, 721% for 2 min and 254% for 48 min of nicotine exposure. Our results indicate that zinc-potentiating effects on alpha4beta4 nAChRs strongly depend on both concentration and time of exposure to nicotine, suggesting that zinc potentiation depends on the degree of desensitization. PMID- 19285093 TI - Expression levels of the alpha4 subunit of the GABA(A) receptor in differentiated neuroblastoma cells are correlated with GABA-gated current. AB - The alpha4 subunit of the GABA(A) receptor (GABAR) is capable of rapid plasticity, increased by chronic exposure to positive GABA modulators, such as the neurosteroid 3alpha-OH-5alpha[beta]-pregnan-20-one (THP). Here, we show that 48 h exposure of differentiated neuroblastoma cells (IMR-32) to 100 nM THP increases alpha4 expression, without changing the current density or the concentration-response curve. Increased expression of alpha4-containing GABAR was verified by a relative insensitivity of GABA (EC(20))-gated current to modulation by the benzodiazepine (BZ) lorazepam (0.01-100 microM), and potentiation of current by flumazenil and RO15-4513, characteristic of alpha4betagamma2 pharmacology. In contrast to THP, compounds which decrease GABA-gated current, such as the BZ inverse agonist DMCM, the GABAR antagonist gabazine and the open channel blocker penicillin, decreased alpha4 expression after a 48 h exposure, without changing BZ responsiveness. However, pentobarbital, another positive GABA modulator, increased alpha4 expression, while the BZ antagonist flumazenil had no effect. In order to test whether changes in current were responsible for increased alpha4 expression, decreases in the Cl(-) driving force were produced by chronic exposure to the NKCC1 blocker bumetanide (10 microM). When applied under these conditions of reduced GABA-gated current, THP failed to increase alpha4 expression. The results of this study suggest that alpha4 expression is correlated with changes in GABA-gated current, rather than simply through ligand receptor interactions. These findings have relevance for GABAR subunit plasticity produced by fluctuations in endogenous steroids across the menstrual cycle, when altered BZ sensitivity is reported. PMID- 19285094 TI - Downregulation of LRP1 [correction of LPR1] at the blood-brain barrier in streptozotocin-induced diabetic mice. AB - Deposition of amyloid-beta peptide (Abeta) in the diabetic brain is poorly understood. Low-density lipoprotein receptor related protein 1(LRP1) at the blood brain barrier (BBB) is critical for regulation of Abeta homeostasis in the brain. In this study, we used streptozotocin-induced diabetic mice to observe the expression of LRP1 at the BBB by Western blot and immunocytochemical analysis, and to study in vivo brain-to-blood efflux transport of 125I-Abeta1-40 using brain clearance studies. In the diabetic mice with hyperglycemia (>16.0 mmol/l) at 6 weeks, LRP1 expression at the BBB was significantly downregulated; no significant changes of LRP1 levels were found at 1 and 3 weeks after diabetes induction. The data of brain clearance studies for Abeta showed significant decrease in LRP1-dependent transport of Abeta across the BBB at 6 weeks after diabetes induction, while no significant changes of LRP1-dependent transport of Abeta across the BBB at 1 or 3 weeks after diabetes induction were apparent. We conclude that the downregulation of LRP1 at the BBB contributes to cerebral Abeta deposition in diabetes mellitus. PMID- 19285095 TI - TGF-beta as a promising option in the treatment of multiple sclerosis. AB - Transforming growth factor-beta (TGF-beta) is a potent regulatory cytokine with diverse effects on hemopoietic cells. The pivotal function of TGF-beta in the immune system is to maintain tolerance via the regulation of lymphocyte proliferation, differentiation, and survival. Among T cells, CD4+CD25+FOXP3+ T regs contain the main source of TGF-beta that suppresses immune responses in inflammatory sites. Defects in TGF-beta1 expression or its signaling in T cells correlate with the onset of several autoimmune diseases. Thus, understanding the function and regulation of TGF-beta during immune responses offers therapeutic promise for the control of autoimmune diseases such as multiple sclerosis. However, the main mechanism by which TGF-beta exerts its protective effects on the experimental model of multiple sclerosis remains to be elucidated. Paradoxically, TGF-beta1 also acts as a pro-inflammatory cytokine and induces interleukin 17-producing pathogenic T helper cells (Th IL-17 cells) synergistically during an inflammatory response in which interleukin 6 is produced. In this review, we will describe the regulatory and therapeutic effects of TGF-beta in multiple sclerosis. PMID- 19285096 TI - Effects of the PPARgamma agonist troglitazone on endothelial cells in vivo and in vitro: differences between human and mouse. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) agonists and PPARgamma/alpha dual agonists have been or are being developed for clinical use in the treatment of type 2 diabetes mellitus and hyperlipidemias. A common tumor finding in rodent carcinogenicity studies for these agonists is hemangioma/hemangiosarcoma in mice but not in rats. We hypothesized that increased endothelial cell proliferation may be involved in the mechanism of PPAR agonist-induced vascular tumors in mice, and we investigated the effects on endothelial cells utilizing troglitazone, the first clinically used PPARgamma agonist, in vivo and in vitro. Troglitazone (400 and 800 mg/kg/day) induced hemangiosarcomas in mice in a 2-year bioassay. We showed that troglitazone increased endothelial cell proliferation in brown and white adipose tissue and liver in mice at sarcomagenic doses after 4 weeks of treatment. Troglitazone was cytotoxic both to human dermal microvascular endothelial cells (HMEC1) and mouse mammary fat pad microvascular endothelial cells (MFP MVEC) at high concentrations. However, MFP MVEC were more resistant to the cytotoxic effects of troglitazone based on the much lower LC(50) in HMEC1 (17.4 muM) compared to MFP MVEC (92.2 muM). Troglitazone increased the proliferation and survival of MFP MVEC but not HMEC1 in growth factor reduced conditions. Our data demonstrate that troglitazone may induce hemangiosarcomas in mice, at least in part, through enhancement of survival and proliferation of microvascular endothelial cells. Such an effect does not occur with human cells, suggesting that human may react differently to exposure to PPAR agonists compared with mice. PMID- 19285097 TI - CYP1A1 and CYP1A2 expression: comparing 'humanized' mouse lines and wild-type mice; comparing human and mouse hepatoma-derived cell lines. AB - Human and rodent cytochrome P450 (CYP) enzymes sometimes exhibit striking species specific differences in substrate preference and rate of metabolism. Human risk assessment of CYP substrates might therefore best be evaluated in the intact mouse by replacing mouse Cyp genes with human CYP orthologs; however, how "human like" can human gene expression be expected in mouse tissues? Previously a bacterial-artificial-chromosome-transgenic mouse, carrying the human CYP1A1_CYP1A2 locus and lacking the mouse Cyp1a1 and Cyp1a2 orthologs, was shown to express robustly human dioxin-inducible CYP1A1 and basal versus inducible CYP1A2 (mRNAs, proteins, enzyme activities) in each of nine mouse tissues examined. Chimeric mice carrying humanized liver have also been generated, by transplanting human hepatocytes into a urokinase-type plasminogen activator(+/+)_severe-combined-immunodeficiency (uPA/SCID) line with most of its mouse hepatocytes ablated. Herein we compare basal and dioxin-induced CYP1A mRNA copy numbers, protein levels, and four enzymes (benzo[a]pyrene hydroxylase, ethoxyresorufin O-deethylase, acetanilide 4-hydroxylase, methoxyresorufin O demethylase) in liver of these two humanized mouse lines versus wild-type mice; we also compare these same parameters in mouse Hepa-1c1c7 and human HepG2 hepatoma-derived established cell lines. Most strikingly, mouse liver CYP1A1 specific enzyme activities are between 38- and 170-fold higher than human CYP1A1 specific enzyme activities (per unit of mRNA), whereas mouse versus human CYP1A2 enzyme activities (per unit of mRNA) are within 2.5-fold of one another. Moreover, both the mouse and human hepatoma cell lines exhibit striking differences in CYP1A mRNA levels and enzyme activities. These findings are relevant to risk assessment involving human CYP1A1 and CYP1A2 substrates, when administered to mice as environmental toxicants or drugs. PMID- 19285098 TI - Transcriptomic configuration of mouse brain induced by adolescent exposure to 3,4 methylenedioxymethamphetamine. AB - The amphetamine derivative (+/-)-3,4-methylenedioxymethamphetamine (MDMA or ecstasy) is a synthetic amphetamine analogue used recreationally to obtain an enhanced affiliative emotional response. MDMA is a potent monoaminergic neurotoxin with the potential to damage brain serotonin and/or dopamine neurons. As the majority of MDMA users are young adults, the risk that users may expose the fetus to MDMA is a concern. However, the majority of studies on MDMA have investigated the effects on adult animals. Here, we investigated whether long term exposure to MDMA, especially in adolescence, could induce comprehensive transcriptional changes in mouse brain. Transcriptomic analysis of mouse brain regions demonstrated significant gene expression changes in the cerebral cortex. Supervised analysis identified 1028 genes that were chronically dysregulated by long-term exposure to MDMA in adolescent mice. Functional categories most represented by this MDMA characteristic signature are intracellular molecular signaling pathways of neurotoxicity, such as, the MAPK signaling pathway, the Wnt signaling pathway, neuroactive ligand-receptor interaction, long-term potentiation, and the long-term depression signaling pathway. Although these resultant large-scale molecular changes remain to be studied associated with functional brain damage caused by MDMA, our observations delineate the possible neurotoxic effects of MDMA on brain function, and have therapeutic implications concerning neuro-pathological conditions associated with MDMA abuse. PMID- 19285099 TI - Minocycline attenuates experimental colitis in mice by blocking expression of inducible nitric oxide synthase and matrix metalloproteinases. AB - In addition to its antimicrobial activity, minocycline exerts anti-inflammatory effects in several disease models. However, whether minocycline affects the pathogenesis of inflammatory bowel disease has not been determined. We investigated the effects of minocycline on experimental colitis and its underlying mechanisms. Acute and chronic colitis were induced in mice by treatment with dextran sulfate sodium (DSS) or trinitrobenzene sulfonic acid (TNBS), and the effect of minocycline on colonic injury was assessed clinically and histologically. Prophylactic and therapeutic treatment of mice with minocycline significantly diminished mortality rate and attenuated the severity of DSS-induced acute colitis. Mechanistically, minocycline administration suppressed inducible nitric oxide synthase (iNOS) expression and nitrotyrosine production, inhibited proinflammatory cytokine expression, repressed the elevated mRNA expression of matrix metalloproteinases (MMPs) 2, 3, 9, and 13, diminished the apoptotic index in colonic tissues, and inhibited nitric oxide production in the serum of mice with DSS-induced acute colitis. In DSS-induced chronic colitis, minocycline treatment also reduced body weight loss, improved colonic histology, and blocked expression of iNOS, proinflammatory cytokines, and MMPs from colonic tissues. Similarly, minocycline could ameliorate the severity of TNBS-induced acute colitis in mice by decreasing mortality rate and inhibiting proinflammatory cytokine expression in colonic tissues. These results demonstrate that minocycline protects mice against DSS- and TNBS-induced colitis, probably via inhibition of iNOS and MMP expression in intestinal tissues. Therefore, minocycline is a potential remedy for human inflammatory bowel diseases. PMID- 19285100 TI - Immune recognition of BoNTs A and B: how anti-toxin antibodies that bind to the heavy chain obstruct toxin action. AB - We localized the BoNT regions that bind blocking Abs from 28 BoNT/A- and 30 BoNT/B-treated dystonia patients who became unresponsive to, and whose sera protected mice against LD100 of, the correlate BoNT. We analyzed Ab binding to BoNT/A- and BoNT/B-peptide panels, each of which consisted of 60, 19-residue peptides that overlapped consecutively by 5 residues and covered the entire H chain of the correlate toxin. Abs bound to a limited set of peptides but levels varied with patient, consistent with responses to each epitope being under separate MHC control. BoNT/B-treated patients had higher anti-toxin Ab levels and bound more H regions (at least 11) than BoNT/A-treated patients (5 regions). The epitopes were on surface areas that did not correlate with surface electrostatic potential, hydrophilicity, hydrophobicity, or temperature factor. Some epitopes within the two toxins display substantial homology and occupy equivalent 3-D locations, occasionally showing a small shift relative to one another, consistent with recognition adjustments accommodating structural differences between the two BoNTs. Blocking Abs bound to BoNT/A at sites that coincided or overlapped with those involved in synaptosome-binding, thus preventing its binding and blocking its entry into the neuron. On BoNT/B, Ab-binding regions overlapped with the sites that bind to mouse and rat synaptotagmin II or to ganglioside, thereby explaining Ab blocking of BoNT/B action. PMID- 19285101 TI - Use of focus groups to understand African-Americans' dietary practices: Implications for modifying a food frequency questionnaire. AB - OBJECTIVE: To generate information about dietary practices, food preferences and food preparation methods from African-Americans in Macon County, Alabama, as a precursor to an intervention designed to modify an existing dietary health questionnaire (DHQ). METHOD: African-American males (30) and females (31) ages 20 to 75 years participated in eight focus groups in Macon County Alabama between June and July, 2007. RESULTS: The core topics identified were dietary practices; food preferences; food preparation methods; fast food practices; and seasonal/specialty foods. The younger focus group participants reported consuming mostly fast foods such as hamburgers for lunch. Fruits, vegetables, salads, fish, chicken and sandwiches were the most common lunch foods for the older males and females. Across the groups, rice, cornbread and potatoes were reportedly the most commonly consumed starchy foods at dinner. Frying and baking were the most common cooking methods. Fewer participants reported removing the skin when cooking chicken versus those who did not remove. Traditional foods including fried green tomatoes and cracklings were selected for addition to the modified DHQ, while those not commonly consumed, were deleted. CONCLUSIONS: Participants described high-fat traditional food preferences, common frying and addition of salted meats to vegetables, which informed the modification of a DHQ. PMID- 19285102 TI - Lower neighbourhood walkability and longer distance to school are related to physical activity in Belgian adolescents. AB - OBJECTIVE: To investigate whether adolescents living in a high-walkable town centre are more physically active than those living in a less-walkable suburb. METHODS: This cross-sectional study was conducted in Izegem (Belgium). Data collection took place in spring 2007. One high-walkable urban neighbourhood and one less-walkable suburban neighbourhood were selected, based on objective connectivity and residential density. One hundred twenty adolescents (12-18 years, 60 per neighbourhood) completed the Neighbourhood Environmental Walkability Scale (NEWS), the Neighbourhood Physical Activity Questionnaire (NPAQ), wore a pedometer for 7 days and filled in an activity log. RESULTS: In contrast with the expectations, adolescents living in the less-walkable suburb reported 220 min/week more cycling for transport than those living in the high walkable town centre. A trend towards significance was found for mean step counts/day with 1371 more steps/day for suburban adolescents. Travel time to school was 7.4 min less for urban adolescents. CONCLUSIONS: In contrast with previous results in adults, lower walkability and larger distance to school was associated with more physical activity in Belgian adolescents. Therefore, physical environmental interventions designed for adults, focusing on increases in connectivity, residential density and connectivity, might not be effective for Belgian adolescents. PMID- 19285103 TI - Advancing the hypothesis that geographic variations in risk factors contribute relatively little to observed geographic variations in heart disease and stroke mortality. AB - PURPOSE: Geographic variation in risk factors may underlie geographic disparities in coronary heart disease (CHD) and stroke mortality. METHODS: Framingham CHD Risk Score (FCRS) and Stroke Risk Score (FSRS) were calculated for 25,770 stroke free and 22,247 CHD-free participants from the REasons for Geographic And Racial Differences in Stroke cohort. Vital statistics provided age-adjusted CHD and stroke mortality rates. In an ecologic analysis, the age-adjusted, race-sex weighted, average state-level risk factor levels were compared to state-level mortality rates. RESULTS: There was no relationship between CHD and stroke mortality rates (r=0.04; p=0.78), but there was between CHD and stroke risk scores at the individual (r=0.68; p<0.0001) and state (r=0.64, p<0.0001) level. There was a stronger (p<0.0001) association between state-level FCRS and state level CHD mortality (r=0.28, p=0.18), than between FSRS and stroke mortality (r=0.12, p=0.56). CONCLUSIONS: Weak associations between CHD and stroke mortality and strong associations between CHD and stroke risk scores suggest that geographic variation in risk factors may not underlie geographic variations in stroke and CHD mortality. The relationship between risk factor scores and mortality was stronger for CHD than stroke. PMID- 19285104 TI - Food insecurity and dyslipidemia among adults in the United States. AB - OBJECTIVE: The objective of this study is to estimate the likelihood of dyslipidemia among food insecure men and women. METHOD: Men, n=2572 and women, n=2977, in the National Health and Nutrition Examination Survey 1999-2002 cholesterol screening sample were included in this study. Gender-stratified descriptive comparisons and logistic regression models were used to study associations between food insecurity and dyslipidemia indicated by abnormal levels of fasting serum triglyceride (TRG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and their ratios. RESULTS: Food insecurity did not associate with dyslipidemia among men. Among women, the associations between food insecurity and dyslipidemia were not consistent. Compared with the fully food secure, women who were marginally food secure were more likely to have abnormal levels of LDL-C (adjusted OR, 1.85; P=0.045) and TRG/HDL-C ratio (adjusted OR, 1.91; P=0.046). Women who were food insecure without hunger were more likely to have abnormal levels of TRG (adjusted OR, 1.90; P=0.041). CONCLUSION: Intermediate-level food insecurity associated with some indicators of dyslipidemia among women but not among men. This observation shows food insecure women may be at risk of dyslipidemia. PMID- 19285105 TI - Regulation of cell proliferation by NADPH oxidase-mediated signaling: potential roles in tissue repair, regenerative medicine and tissue engineering. AB - The superoxide generating enzyme NADPH oxidase has received much attention as a major cause of oxidative stress underlying vascular disease. However, there is increasing evidence that oxidant signaling involving NADPH oxidase has other important roles in cell biology. Nox family proteins are the catalytic, electron transporting subunits of the NADPH oxidase enzyme complex. It is now clear that reactive oxygen species (ROS) generated by NADPH oxidase participate in intracellular signaling processes that regulate cell differentiation and proliferation. These mechanisms are important in tissue repair and tumorigenesis, diverse conditions where cell proliferation is required, but when poorly controlled the generation of ROS is obviously detrimental. Indeed, NADPH oxidase mediated cell proliferation has been observed in a wide range of cell types including those found in blood vessels, kidney, liver, skeletal muscle precursors, neonatal cardiac myocytes, lung epithelial cells, gastric mucosa, brain microglia, and a variety of cancer cells. NADPH oxidases act not as isolated elements downstream of a particular pathway, but rather may amplify multiple receptor tyrosine kinase-mediated processes by inhibiting protein tyrosine phosphatases. Therefore, NADPH oxidase-mediated redox signaling may represent a unique intracellular amplifier of diverse signaling pathways involved in tissue repair processes such as cell proliferation, wound healing, angiogenesis and fibrosis. Recent studies also suggest that NADPH oxidase is involved in differentiation of stem cells. As occurs in unresolved inflammation, however, hyperactivity of this enzyme system leads to tissue injury. Thus modulating NADPH oxidase may have significant impacts on regenerative medicine and tissue engineering, such as growing heart muscle. PMID- 19285106 TI - Value judgments and self-control of action: the role of the medial frontal cortex. AB - Humans generate actions in relation to perceived events in the environment. Events are valuated in terms of subjective (personal) relevance or meaning, i.e. "what does this mean to me?". Similarly, making sense or gaining meaning from sensations (i.e., "perception") from one's own body and of mental images, such as memories or intentions, involves valuation from a subjective perspective. Here, we review recent findings in neurophysiology and neuroimaging suggesting that the medial frontal cortex comprises cortical relay nodes that afford the attribution of self-relevant, immediate and intuitive (implicit) meaning. In addition, we describe recent data that suggest that the medial frontal cortex participates also in the explicit appraisal of certain stimuli, namely, emotional face expressions, occurring as early as 150 ms following the stimulus. We propose that the medial frontal cortex subserves egocentric "value" judgments (both implicit and explicit), which are critical for self-control of action. PMID- 19285107 TI - A co-printed oligomer to enhance reliability of spotted microarrays. AB - Successful printing and hybridization is essential for efficient and reliable data acquisition in a spotted microarray experiments. In this study we demonstrated that printing a 25 mer (printed 25 mer) with a standard 70 mer probe in each spot followed by the use of a fluorescently labeled 25 mer complement in the hybridization mixture ensures monitoring overall printing quality of the chip. This system can also be used as a control to evaluate adequate hybridization, washing, and alignment of spots to position the tracking grids during scanning. A print correction value incorporated in data analysis enhances consistency and reliability of results. PMID- 19285109 TI - Albumin nanoparticles targeted with Apo E enter the CNS by transcytosis and are delivered to neurones. AB - The blood-brain barrier (BBB) represents a considerable obstacle to brain entry of the majority of drugs and thus severely restricts the therapy of many serious CNS diseases including brain tumours, brain HIV, Alzheimer and other neurodegenerative diseases. The use of nanoparticles coated with polysorbate 80 or with attached apolipoprotein E has enabled the delivery of drugs across the BBB. However, the mechanism of this enhanced transport is still not fully understood. In this present study, human serum albumin nanoparticles, with covalently bound apolipoprotein E (Apo E) as a targetor as well as without apolipoprotein E, were manufactured and injected intravenously into SV 129 mice. The animals were sacrificed after 15 and 30 min, and their brains were examined by transmission electron microscopy. Only the nanoparticles with covalently bound apolipoprotein E were detected in brain capillary endothelial cells and neurones, whereas no uptake into the brain was detectable with nanoparticles without apolipoprotein E. We have also demonstrated uptake of the albumin/ApoE nanoparticles into mouse endothelial (b.End3) cells in vitro and their intracellular localisation. These findings indicate that nanoparticles with covalently bound apolipoprotein E are taken up into the cerebral endothelium by an endocytic mechanism followed by transcytosis into brain parenchyma. PMID- 19285108 TI - A longitudinal study in youth of heart rate variability at rest and in response to stress. AB - BACKGROUND: Few longitudinal studies have examined ethnic and sex differences, predictors and tracking stabilities of heart rate variability (HRV) at rest and in response to stress in youths and young adults. METHODS: Two evaluations were performed approximately 1.5 years apart on 399 youths and young adults (189 European Americans [EAs] and 210 African Americans [AAs]; 190 males and 209 females). HRV was measured at rest and during a video game challenge. RESULTS: AAs showed significantly higher resting root mean square of successive differences (RMSSD) of normal R-R intervals and high-frequency (HF) power than EAs (ps<0.01). Females displayed larger decrease of RMSSD and HF during video game challenge than males (ps<0.05). These ethnic and sex differences were consistent across 1.5 years. No significant sex difference of resting HRV or ethnic difference of HRV response to stress was observed. In addition to age, ethnicity or sex, baseline resting HRV or HRV response to stress are predictors of the corresponding variables 1.5 years later (ps<0.01). Furthermore, weight gain indexed by either body mass index or waist circumference predicts declined resting HRV levels during follow up (ps<0.05). Tracking stabilities were high (>0.5) for resting HRV, but relatively low (<0.3) for HRV in response to stress. CONCLUSION: AAs show higher resting HRV than EAs, and females display greater HRV response to stress than males; and these ethnic and sex differences are consistent across 1.5 years. Resting HRV declines with weight gain. PMID- 19285110 TI - Antidepressant-like synergism of extracts from magnolia bark and ginger rhizome alone and in combination in mice. AB - Magnolia bark and ginger rhizome is a drug pair in many prescriptions for treatment of mental disorders in traditional Chinese medicine (TCM). However, compatibility and synergism mechanism of two herbs on antidepressant actions have not been reported. The aim of this study was to approach the rationale of the drug pair in TCM. We evaluated antidepressant-like effects of mixture of honokiol and magnolol (HMM), polysaccharides (PMB) from magnolia bark, essential oil (OGR) and polysaccharides (PGR) from ginger rhizome alone, and the possibility of synergistic interactions in their combinations in the mouse forced swimming test (FST) and tail suspension test (TST). Serotonin (5-HT) and noradrenaline (NE) levels in prefrontal cortex, hippocampus and striatum were also examined. 30 mg/kg HMM decreased immobility in the FST and TST in mice after one- and two-week treatment. OGR (19.5 or 39 mg/kg) alone was ineffective. The combination of an ineffective dose of 39 mg/kg OGR with 15 mg/kg HMM was the most effective and produced a synergistic action on behaviors after two-week treatment. Significant increase in 5-HT and synergistic increase in NE in prefrontal cortex were observed after co-administration of HMM with OGR. These results demonstrated that HMM was the principal component of this drug pair, whereas OGR served as adjuvant fraction. Compatibility of HMM with OGR was suggested to exert synergistic antidepressant actions by attenuating abnormalities in serotonergic and noradrenergic system functions. Therefore, we confirmed the rationality of drug pair in clinical application and provided a novel perspective in drug pair of TCM researches. PMID- 19285111 TI - C957T polymorphism of the human dopamine D2 receptor gene predicts extrastriatal dopamine receptor availability in vivo. AB - The C957T (rs6277) single nucleotide polymorphism (SNP) of the human dopamine D2 receptor (DRD2) gene (DRD2) affects DRD2 mRNA stability and has been shown to predict striatal DRD2 availability (B(max)/K(D)) in vivo in man. Specifically, the C/C genotype is associated with low striatal DRD2 availability (C/CC/T>T/T). Also the TaqIA A1 allele carriers (p=0.101) tended to have higher extrastriatal DRD2 BP(ND) compared to non-carriers whereas the -141C Ins/Del genotype did not influence extrastriatal DRD2 BP(ND). Our findings indicate that the DRD2 SNPs regulate DRD2 availability in the human cortex and in the thalamus in vivo. However, the regulation pattern is different from that observed previously for striatal DRD2 availability in vivo, which may reflect distinct functional roles of dopamine and DRD2 in the cortex versus the striatum. The results provide useful information for the interpretation of genetic studies exploring the role of the DRD2 in normal physiology as well as in psychiatric and neurological diseases. PMID- 19285112 TI - Selenoprotein function and muscle disease. AB - The crucial role of the trace element selenium in livestock and human health, in particular in striated muscle function, has been well established but the underlying molecular mechanisms remain poorly understood. Over the last decade, identification of the full repertoire of selenium-containing proteins has opened the way towards a better characterization of these processes. Two selenoproteins have mainly been investigated in muscle, namely SelW and SelN. Here we address their involvement in muscle development and maintenance, through the characterization of various cellular or animal models. In particular, mutations in the SEPN1 gene encoding selenoprotein N (SelN) cause a group of neuromuscular disorders now referred to as SEPN1-related myopathy. Recent findings on the functional consequences of these mutations suggest an important contribution of SelN to the regulation of oxidative stress and calcium homeostasis. Importantly, the conclusions of these experiments have opened new avenues of investigations that provide grounds for the development of therapeutic approaches. PMID- 19285113 TI - Blocking central leukotrienes synthesis affects vasopressin release during sepsis. AB - Recent studies revealed that vasopressinergic neurons have a high content of cys leukotriene C(4) (LTC(4)) synthase, a critical enzyme in cys-leukotriene synthesis that may play a role in regulating vasopressin secretion. This study investigates the role of this enzyme in arginine vasopressin (AVP) release during experimentally induced sepsis. Male Wistar rats received an i.c.v. injection of 3 [1-(p-chlorobenzyl)-5-(isopropyl)-3-tert-butylthioindol-2-yl]-2, 2 dimethylpropanoic acid (MK-886) (1.0 microg/kg), a leukotrienes (LTs) synthesis inhibitor, or vehicle, 1 h before cecal ligation and puncture (CLP) or sham operation. In one group of animals the survival rate was monitored for 3 days. In another group, the animals were decapitated at 0, 4, 6, 18 and 24 h after CLP or sham operation, and blood was collected for hematocrit, serum sodium and nitrate, plasma osmolality, protein and AVP determination. A third group was used for blood pressure measurements. The neurohypophysis was removed for quantification of AVP content, and the hypothalamus was dissected for LTC(4) synthase analysis by Western blot. Mortality after CLP was reduced by the central administration of MK-886. The increase in plasma AVP levels and hypothalamus LTC(4) synthase content in the initial phase of sepsis was blocked, whereas the decrease in neurohypophyseal AVP content was partially reversed. Also the blood pressure drop was abolished in this phase. The increase of serum nitric oxide and hematocrit was reduced, and the decrease in plasma protein and osmolality was not affected by the LTs blocker. In the final phase of sepsis, the plasma AVP level and the hypothalamic LTC(4) synthase content were at basal levels. The central administration of MK-886 increased the hypothalamic LTC(4) synthase content but did not alter the plasma and neurohypophysis AVP levels observed, or the blood pressure during this phase. These results suggest that the central LTs are involved in the vasopressin release observed during sepsis. PMID- 19285114 TI - Bright light produces Fos-positive neurons in caudal trigeminal brainstem. AB - Excessive discomfort after exposure to bright light often occurs after ocular injury and during headache. Although the trigeminal nerve is necessary for light evoked discomfort, the mechanisms underlying this phenomenon, often referred to generally as photophobia, are not well defined. Quantitative Fos-like immunoreactivity (Fos-LI) was used to determine the pattern of neuronal activation in the caudal brainstem after bright light stimulation and, secondly, whether a neurovascular mechanism within the eye contributes to this response. Under barbiturate anesthesia, male rats were exposed to low (1 x 10(4) lx) or high intensity (2 x 10(4) lx) light delivered from a thermal neutral source for 30 min (30 s ON, 30 s OFF) and allowed to survive for 90 min. Intensity-dependent increases in Fos-LI were seen in laminae I-II at the trigeminal caudalis/cervical cord junction region (Vc/C1) and nucleus tractus solitarius (NTS). Fos-LI also increased at the trigeminal interpolaris/caudalis transition (Vi/Vc(vl)) and dorsal paratrigeminal (dPa5) regions independent of intensity. Intravitreal injection of norepinephrine greatly reduced light-evoked Fos-LI at the Vc/C1, dPa5 and NTS, but not at the Vi/Vc transition. Lidocaine applied to the ocular surface had no effect on Fos-LI produced in trigeminal brainstem regions. These results suggested that multiple regions of the caudal trigeminal brainstem complex integrate light-related sensory information. Fos-LI produced at the dPa5 and NTS, coupled with norepinephrine-induced inhibition, was consistent with the hypothesis that light-evoked activation of trigeminal brainstem neurons involves an intraocular neurovascular mechanism with little contribution from neurons that supply the ocular surface. PMID- 19285115 TI - Molecular and functional interactions between tumor necrosis factor-alpha receptors and the glutamatergic system in the mouse hippocampus: implications for seizure susceptibility. AB - Tumor necrosis factor (TNF)-alpha is a proinflammatory cytokine acting on two distinct receptor subtypes, namely p55 and p75 receptors. TNF-alpha p55 and p75 receptor knockout mice were previously shown to display a decreased or enhanced susceptibility to seizures, respectively, suggesting intrinsic modifications in neuronal excitability. We investigated whether alterations in glutamate system function occur in these naive knockout mice with perturbed cytokine signaling that could explain their different propensity to develop seizures. Using Western blot analysis of hippocampal homogenates, we found that p55(-/-) mice have decreased levels of membrane GluR3 and NR1 glutamate receptor subunits while GluR1, GluR2, GluR6/7 and NR2A/B were unchanged as compared to wild-type mice. In p75(-/-) mice, GluR2, GluR3, GluR6/7 and NR2A/B glutamate receptor subunits were increased in the hippocampus while GluR1 and NR1 did not change. Extracellular single-cell recordings of the electrical activity of hippocampal neurons were carried out in anesthetized mice by standard electrophysiological techniques. Microiontophoretic application of glutamate increased the basal firing rate of hippocampal neurons in p75(-/-) mice versus wild-type mice, and this effect was blocked by 2-amino-5-phosphopentanoic acid and 6-nitro-7-sulfamoyl benzo(f)quinoxaline-2,3-dione denoting the involvement of N-methyl-D-aspartic acid and AMPA receptors. In p55(-/-) mice, hippocampal neurons responses to glutamate were similar to wild-type mice. Spontaneous glutamate release measured by in vivo hippocampal microdialysis was significantly decreased only in p55(-/-) mice. No changes were observed in KCl-induced glutamate release in both receptor knockout mice strains versus wild-type mice. These findings highlight specific molecular and functional interactions between p55 and p75 receptor-mediated signaling and the glutamate system. These interactions may be relevant for controlling neuronal excitability in physiological and pathological conditions. PMID- 19285116 TI - Parvalbumin-immunoreactive neurons and GABAergic neurons of the basal forebrain project to the rat basolateral amygdala. AB - The basal forebrain (BF) contains a diffuse array of cholinergic and non cholinergic neurons that project to the cerebral cortex and basolateral nuclear complex of the amygdala (BLC). Previous studies have shown that the GABAergic subpopulation of non-cholinergic corticopetal BF neurons selectively innervates cortical interneurons. Although several investigations in both rodents and primates have indicated that some BF neurons projecting to the BLC are non cholinergic, there have been no studies that have attempted to identify the neurochemical phenotype(s) of these neurons. The present study combined Fluorogold retrograde tract tracing with immunohistochemistry for two markers of BF GABAergic neurons, parvalbumin (PV) or glutamic acid decarboxylase (GAD), to determine if a subpopulation of BF GABAergic cells projects to the BLC. Injections of Fluorogold confined to the rat BLC, and centered in the basolateral nucleus, produced extensive retrograde labeling in the ventral pallidum and substantia innominata regions of the BF. Although the great majority of retrogradely labeled neurons were not double-labeled, about 10% of these neurons, located mainly along the ventral aspects of the fundus striati and globus pallidus, exhibited immunoreactivity for PV or GAD. The results of this investigation contradict the long-held belief that there is no extra-amygdalar source of GABAergic inputs to the BLC, and indicate that the cortex-like BLC, in addition to the cortex proper, receives inhibitory inputs from the basal forebrain. PMID- 19285117 TI - Selective activation of nuclear factor kappa B in the cochlea by sensory and inflammatory stress. AB - Damage response pathways triggered by mechanical stress might reasonably be expected to be conserved throughout evolution. However, using a nuclear factor kappa B (NF-kappaB) reporter mouse we show here that this phylogenetically recent transcription factor plays a major role in the response to mechanosensory stress in the mammalian inner ear. The protective action of NF-kappaB is exerted in neither sensory nor non-sensory epithelial cells, but rather in connective tissue cells within the spiral ligament and spiral limbus. In the spiral ligament, predominantly type I fibrocytes are activated following noise exposure, whereas type II fibrocytes are activated following systemic inflammatory stress. Immune mediated and acoustic trauma-mediated hearing loss syndromes in humans may in part result from the vulnerability of type II and type I fibrocytes to systemic inflammatory stress and acoustic trauma, respectively. Unexpected cell-specific and stress-specific NF-kappaB activation found in the inner ear in this in vivo study suggest that this approach may have wide applications in demonstrating similar specializations of stress responses in other tissues, including the brain. PMID- 19285118 TI - Cerebellar white matter injury following systemic endotoxemia in preterm fetal sheep. AB - Injury to the cerebellum and brainstem is becoming increasingly recognized in prematurely born infants. The role of infection/inflammation in mediating damage to those structures in the preterm brain is largely unknown. Preterm fetal sheep (70% gestation) received either saline-vehicle (control group; n=11) or Escherichia coli lipopolysaccharide (100 ng intravenous [i.v.]; lipopolysaccharide [LPS] group; n=9), and were allowed to recover for 3 days before sacrifice. A diffuse pattern of cerebellar white matter damage was observed in all animals exposed to LPS, while focal cerebellar white matter lesions were observed in three out of nine animals, and an intragyral white matter hemorrhage in one animal. Cerebellar white matter injury was associated with a statistically significant loss of oligodendrocyte transcription factor-2 positive oligodendrocytes and increased terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling-positive cell counts. Ionized calcium binding adapter molecule 1 (Iba1)-positive cells which had the morphology of activated microglia were commonly observed in areas of injury. There was no obvious injury to the cerebellar cortex or to cerebellar Purkinje cells, and no obvious injury in any region of the brainstem. These data provide support for a role of infection/inflammation in selective white matter injury in the immature cerebellum, and demonstrate a differential vulnerability of the brainstem and cerebellar white matter to injury at this time. PMID- 19285119 TI - Glial cell line-derived neurotrophic factor family ligands enhance capsaicin stimulated release of calcitonin gene-related peptide from sensory neurons. AB - The glial cell line-derived neurotrophic factor (GDNF) family ligands (GFLs) are a group of peptides that have been implicated as important factors in inflammation, since they are released in increased amounts during inflammation and induce thermal hyperalgesia upon injection. Mouse isolated sensory neurons in culture and freshly dissociated spinal cord slices were used to examine the enhancement in stimulated-release of the neuropeptide, calcitonin gene-related peptide (CGRP), as a measure of sensitization. Exposure of isolated sensory neurons in culture to GDNF, neurturin, and artemin enhanced the capsaicin stimulated release of immunoreactive calcitonin gene-related peptide (iCGRP) two- to threefold, but did not increase potassium-stimulated release of iCGRP. A similar profile of sensitization was observed in freshly dissociated spinal cord slices. Persephin, another member of the GFL family thought to be important in development, was unable to induce an enhancement in the release of iCGRP. These results demonstrate that specific GFLs are important mediators affecting sensory neuronal sensitivity, likely through modulation of the capsaicin receptor. The sensitization of sensory neurons during inflammation, and the pain and neurogenic inflammation resulting from this sensitization, may be due in part to the effects of these selected GFLs. PMID- 19285120 TI - Stress axis plasticity during vestibular compensation in the adult cat. AB - The postural, ocular motor, perceptive and neurovegetative syndromes resulting from unilateral vestibular neurectomy (UVN) symptoms could generate a stress and thereby activate the hypothalamo-pituitary-adrenal (HPA) axis. This study was aimed at determining whether UVN causes changes in the activity of the HPA axis, and if so, evaluating the time course of changes associated with UVN syndrome. At the cellular level, corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) immunoreactivity (Ir) were analyzed and quantified in the paraventricular nucleus (PVN) and the vestibular nuclei (VN) complex of cats killed early (1 and 7 days) or late (30 and 90 days) after UVN. Dopamine-beta-hydroxylase (DbetaH), the enzyme synthesizing noradrenaline was examined in the locus coeruleus (LC) in these same cats. At the behavioral level, the time course of recovery of the postural and locomotor functions was quantified at the same postoperative delays in another group of UVN cats. Results showed a significant bilateral increase in the number of both AVP-Ir and CRF-Ir neurons in the PVN and an increase of DbetaH Ir neurons in the LC at 1, 7 and 30 days after UVN. This increased number of neurons was no longer observed at 90 days. Conversely, a significant bilateral decrease of CRF-Ir neurons was observed in the VN at these same postlesion times, with a similar return to control values at 90 days. Our behavioral observations showed strong posturo-locomotor functional deficits early after UVN (1 and 7 days), which had recovered partially at 30 days and completely by 90 days postlesion. We demonstrate a long-lasting activation of the HPA axis, which likely reflects a chronic stress, experienced by the animals, which corresponds to the time course of full vestibular compensation, and which is no longer present when the animals are completely free of posturo-locomotor symptoms at 90 days. PMID- 19285121 TI - Early dysfunction of central 5-HT system in a murine model of bovine spongiform encephalopathy. AB - The hypothesis of an early vulnerability of the serotonergic system to prion infection was investigated in a murine model of bovine spongiform encephalopathy (BSE). Behavioral tests targeted to 5-HT functions were performed in the course of infection to evaluate circadian activity, anxiety-like behavior, pain sensitivity and the 5-HT syndrome. The first behavioral change was a decrease in nocturnal activity detected at 30% of incubation time. Further behavioral alterations including nocturnal hyperactivity, reduced anxiety, hyperalgesia and exaggerated 5-HT syndrome were observed at 60%-70% of incubation time, before the onset of clinical signs. The same tests performed in 5-HT-depleted mice and in prion protein-deficient mice revealed behavioral abnormalities similar in many aspects to those of BSE-infected mice. Histological and biochemical analysis showed alterations of the serotonergic system in BSE-infected and prion protein deficient mice. These results indicate that BSE infection affects the homeostasis of serotonergic neurons and suggest that the disruption of prion protein normal function contributes to the early pathological changes in our mouse model of BSE. A similar process may occur in the human variant Creutzfeldt-Jacob disease, as suggested by the early symptoms of alterations in mood, sleep and pain sensitivity. PMID- 19285122 TI - Juvenile separation stress induces rapid region- and layer-specific changes in S100ss- and glial fibrillary acidic protein-immunoreactivity in astrocytes of the rodent medial prefrontal cortex. AB - The impact of juvenile stress exposure on astrocyte plasticity was assessed in the precocious rodent Octodon degus. Astrocytes expressing S100ss and glial fibrillary acidic protein (GFAP) were quantified in the limbic medial prefrontal cortex (mPFC), including the anterior cingulate (ACd), precentral medial (PrCm), infra- (IL) and prelimbic (PL) cortex and in the "non-limbic" somatosensory cortex (SSC). At the age of 21 days we compared (i) controls (C), (ii) stressed animals (SSR: separation stress/short reunion), which were exposed to 6 h separation from the family, followed by 1 h reunion with the family and (iii) stressed animals (SER: separation stress/extended reunion), which were stressed like group SSR but exposed to 48 h reunion. The observed glia response was already measurable 7 h after the onset of the stress exposure. Compared to controls SER and SSR animals showed elevated densities of S100ss-IR astrocytes in layers II/III and V-VI of the ACd, IL and PrCm, whereas no significant group differences were observed in the PL and SSC. The SSR group showed significantly decreased density of GFAP-immunoreactive astrocytes in all mPFC subregions. Only in the ACd the stress-induced changes in glia density were still evident after 48 h reunion with the family. Compared to controls, the length of GFAP-IR processes and the number of ramification points were significantly reduced in all mPFC subregions and in the SSC of the SSR group. In the SSC the stress-evoked changes in GFAP-glia density were opposite compared to the changes seen in the medial prefrontal cortical subregions, whereas the changes in GFAP-labeled processes were comparable to those observed in the mPFC. In summary, these results demonstrate that a single stress episode induces rapid and quite complex region- and cell-specific changes in glial cells, reflected by an upregulation of cytoplasmic (S100ss) and downregulation of cytoskeletal (GFAP) glial protein. PMID- 19285123 TI - Two phenolic glycosides from Curculigo orchioides Gaertn. AB - One new glycoside derivative from syringic acid and one new phenol glycoside, curculigoside E (1) and orchioside D (2), were isolated and characterized from the rootstock of Curculigo orchioides collected in the Nawalparasi District (Nepal). The structures of the new isolated compounds were elucidated by means of spectroscopic methods such as 1D, 2D NMR and MS. PMID- 19285124 TI - The complete mitochondrial genome of Atelura formicaria (Hexapoda: Zygentoma) and the phylogenetic relationships of basal insects. AB - In this study, the complete sequence of the mitochondrial DNA (mtDNA) of Atelura formicaria (Hexapoda: Zygentoma) is described. The molecule is 15.205 bp in length and it is the third complete mt genome sequenced from the Zygentoma. The genome organization conforms with the putative ancestral insect gene arrangement. All protein coding genes use standard initiation codons (methionine and isoleucine). The exception is nad4 that starts with GTG, a codon used for this purpose in other insect species. A peculiar strand skew bias is observed, given that the PCGs encoded on the J-strand contain more thymines than adenines and more cytosines than guanines. This trend in nucleotide composition has been observed also in the "firebrat" Thermobia domestica (Zygentoma, Lepismatidae), but differs from that of the majority of hexapod species, including Tricholepidion gertschi (Zygentoma, Lepidotrichidae), where adenines and cytosines outnumber thymines and guanines. The presence of structural elements in the control region is also discussed, with emphasis on their potential implications in the regulation of replication and/or transcription of the mitochondrial genome. A mitogenomic phylogenetic analysis, centered on the internal relationships within Zygentoma and on the position of Microcoryphia and Zygentoma among Ectognatha, is performed. PMID- 19285125 TI - Functional analysis of the porcine USP18 and its role during porcine arterivirus replication. AB - Emerging evidence places deubiquitylation at the core of a multitude of regulatory processes, ranging from cell growth to innate immune response and health, such as cancer, degenerative and infectious diseases. Little is known about deubiquitylation in pig and arterivirus infection. This report provides information on the biochemical and functional role of the porcine USP18 during innate immune response to the porcine respiratory and reproductive syndrome virus (PRRSV). We have shown that UBP gene is the ortholog of the murine USP18 (Ubp43) gene and the human ubiquitin specific protease 18 (USP18) gene and encodes a biochemically functional de-ubiquitin enzyme which inhibits signalling pathways that lead to IFN-stimulating response element (ISRE) promotor regulation. Furthermore we have demonstrated that overexpression of the porcine USP18 leads to reduced replication and/or growth of PRRSV. Our data contrast with the conclusion of numerous reports demonstrating that USP18-deficient mice are highly resistant to viral and bacterial infections and to oncogenic transformation by BCR-ABL, and highlight USP18 as a potential target gene that promotes reduced replication of PRRSV. PMID- 19285126 TI - Response of mechanosensory hair cells of the zebrafish lateral line to aminoglycosides reveals distinct cell death pathways. AB - We report a series of experiments investigating the kinetics of hair cell loss in lateral line neuromasts of zebrafish larvae following exposure to aminoglycoside antibiotics. Comparisons of the rate of hair cell loss and the differential effects of acute versus chronic exposure to gentamicin and neomycin revealed markedly different results. Neomycin induced rapid and dramatic concentration dependent hair cell loss that is essentially complete within 90 min, regardless of concentration or exposure time. Gentamicin-induced loss of half of the hair cells within 90 min and substantial additional loss, which was prolonged and cumulative over exposure times up to at least 24h. Small molecules and genetic mutations that inhibit neomycin-induced hair cell loss were ineffective against prolonged gentamicin exposure supporting the hypothesis that these two drugs are revealing at least two cellular pathways. The mechanosensory channel blocker amiloride blocked both neomycin and gentamicin-induced hair cell death acutely and chronically indicating that these aminoglycosides share a common entry route. Further tests with additional aminoglycosides revealed a spectrum of differential responses to acute and chronic exposure. The distinctions between the times of action of these aminoglycosides indicate that these drugs induce multiple cell death pathways. PMID- 19285127 TI - Misidentification of nitrated peptides: comments on Hong, S.J., Gokulrangan, G., Schoneich, C., 2007. Proteomic analysis of age-dependent nitration of rat cardiac proteins by solution isoelectric focusing coupled to nanoHPLC tandem mass spectrometry. Exp. Gerontol. 42, 639-651. PMID- 19285128 TI - AspAlt: A tool for inter-database, inter-genomic and user-specific comparative analysis of alternative transcription and alternative splicing in 46 eukaryotes. AB - We have developed AspAlt-a web-based comparative analytical platform for exploring the variations in alternative transcription (AT) events and alternative splicing (AS) events in eukaryotes. AspAlt provides integrated access to 2.1 million AT-AS annotations from 1,58,876 multi-isoform genes and has the following user-friendly analytical features: (1) advanced graphical display to visualize and analyze AT-AS events in 46 eukaryotic genomes; (2) compare and identify the differences in AT-AS patterns among a group of genes specified by the user or among homologous gene groups; (3) inter-database comparative viewer to analyze the differences in the AT-AS annotations for the same gene among Ensembl, RefSeq and AceView databases; (4) dynamically classify and generate graphical plots of AT-AS events from mRNA annotations submitted by the user; and (5) download genomic AT-AS annotations of 46 eukaryotes in XML and tab-delimited formats. The AspAlt resource is available at http://66.170.16.154/AspAlt. PMID- 19285129 TI - Optimizing comparative genomic hybridization probes for genotyping and SNP detection in Plasmodium falciparum. AB - Microarray-based comparative genomic hybridizations (CGH) interrogate genomic DNA to identify structural differences such as amplifications and deletions that are easily detected as large signal aberrations. Subtle signal deviations caused by single nucleotide polymorphisms (SNPs) can also be detected but is challenged by a high AT content (81%) in P. falciparum. We compared genome-wide CGH signal to sequence polymorphisms between parasite strains 3D7, HB3, and Dd2 using NimbleGen microarrays. From 23,191 SNPs (excluding var/rif/stevor genes), our CGH probe set detected SNPs with >99.9% specificity but low (<10%) sensitivity. Probe length, melting temperature, GC content, SNP location in the probe, mutation type, and hairpin structures affected SNP sensitivity. Previously unrecognized variable number tandem repeats (VNTRs) also were detected by this method. These findings will guide the redesign of a probe set to optimize an openly available CGH microarray platform for high-resolution genotyping suitable for population genomics studies. PMID- 19285130 TI - Vitamins E and C in the prevention of cardiovascular disease and cancer in men. PMID- 19285131 TI - Elevated plasma corticosterone level and depressive behavior in experimental temporal lobe epilepsy. AB - Depression is frequently reported in epilepsy patients; however, mechanisms of co morbidity between epilepsy and depression are poorly understood. An important mechanism of depression is disinhibition within the hypothalamo-pituitary adrenocortical (HPA) axis. We examined the functional state of the HPA axis in a rat model of co-morbidity between temporal lobe epilepsy and depression. Epilepsy was accompanied by the interictal elevation of plasma corticosterone, and by the positively combined dexamethasone/corticotropin releasing hormone test. The extent of the HPA hyperactivity was independent of recurrent seizures, but positively correlated with the severity of depressive behavior. We suggest that the observed hyperactivity of the HPA axis may underlie co-morbidity between epilepsy and depression. PMID- 19285132 TI - Neuropeptide Y stimulates proliferation, migration and differentiation of neural precursors from the subventricular zone in adult mice. AB - The neuropeptide Y (NPY) is widely expressed in the central nervous system and has been shown to stimulate neurogenesis in the hippocampus and the olfactory epithelium. Here, we demonstrate that intracerebroventricular injection of NPY stimulates proliferation of neural precursors in the mice subventricular zone (SVZ), one the most neurogenic areas of the brain. Newly generated neuroblasts migrate through the rostral migratory stream to the olfactory bulb and also directly to the striatum, as evidenced by BrdU labelling and cell phenotyping. Using knock-out mice, specific NPY receptor agonists and antagonists, we report that this neuroproliferative effect is mediated by the Y1 receptor subtype that we found to be highly expressed in the SVZ both at the mRNA and protein levels. Our data suggest that stimulating endogenous SVZ neural stem cells by NPY may be of a potential interest in cell replacement based therapies of neurodegenerative diseases affecting the striatum such as Huntington's disease. PMID- 19285133 TI - Increased activation of Iba1+ microglia in pediatric epilepsy patients with Rasmussen's encephalitis compared with cortical dysplasia and tuberous sclerosis complex. AB - Microgliosis is prominent in Rasmussen's encephalitis (RE), a disease with severe seizure activity. However, it is unclear if microglial activation is similar with different histopathologic substrates. Iba1-immunolabelled microglial activation was assessed in neocortex from pediatric epilepsy surgery patients with RE (n=8), cortical dysplasia (CD; n=6) and tuberous sclerosis complex (TSC; n=6). Microglial reactivity was increased, in severely affected RE areas (29% labeling) compared with minimally affected areas of RE cases (15%) and cases of TSC (14%) and CD (12%). There was no qualitative association of Iba1 immunolabelling with the presence of CD8(+) cytotoxic T-cells and no statistical association with clinical epilepsy variables, such as seizure duration or frequency. Iba1 appears to be an excellent marker for detecting extensive microglial activation in patients with RE. In addition, this study supports the notion that Iba1-labeled microglial activation is increased in patients with active RE, compared with cases of CD and TSC. PMID- 19285135 TI - Extracellular GAPDH binds to L1 and enhances neurite outgrowth. AB - We have identified glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a binding partner for the cell adhesion molecule L1. GAPDH binds to sites within the extracellular domain of L1, namely the immunoglobulin-like domains I-VI and the fibronectin type III homologous repeats 4-5. Extracellular GAPDH was detected at the cell surface of neuronal cells by surface biotinylation and immunocytochemistry. Addition of GAPDH antibodies to cultured cerebellar neurons inhibited L1-dependent neurite outgrowth in the presence of ATP, while the application of exogenous GAPDH promoted L1-dependent neurite outgrowth. Pre treatment of substrate-coated L1-Fc with ATP and GAPDH, which phosphorylates L1, subsequently led to an enhanced neurite outgrowth. Furthermore, aggregation of L1 Fc carrying beads was enhanced in the presence of both GAPDH and ATP. L1 dependent neurite outgrowth and aggregation of L1 were diminished in the presence of alkaline phosphatase or a protein kinase inhibitor. Our results show that GAPDH-dependent phosphorylation of L1 is a novel mechanism in regulating L1 mediated neurite outgrowth. PMID- 19285134 TI - Chipping away at diagnostics for neurodegenerative diseases. AB - Biomarkers are needed to overcome critical roadblocks in the development of disease-modifying therapeutics for neurodegenerative diseases. Evolving genome wide expression technologies can comprehensively search for molecular biomarkers and allow fascinating insights into the expanding complexity of the human transcriptome. The technology has matured to the point where some applications are deemed reliable enough for use in patient care. In the neurosciences, it has led to the discoveries of osteopontin in multiple sclerosis and SORL1/LR11 in Alzheimer's, and recent studies indicate its potential for identifying neurogenomic biomarkers. Advances in pre-analytical and analytical methods are improving search efficiency and reproducibility and may lead to a pipeline of biomarker candidates suitable for development into future neurologic diagnostics. PMID- 19285136 TI - Macrophage stimulating protein is a neurotrophic factor for a sub-population of adult nociceptive sensory neurons. AB - Macrophage stimulating protein (MSP) is a pleiotropic growth factor that signals via the RON receptor tyrosine kinase. Here we demonstrate that MSP increases the proportion of cultured adult mouse DRG neurons displaying discernable neuritic processes and promotes the elongation and branching of these processes in a dose dependent manner. RON expression in adult DRG is largely restricted to nerve growth factor (NGF)-responsive nociceptive neurons, and MSP mimics the effects of NGF by increasing the expression of several mRNAs that encode functionally important proteins that are characteristically expressed by this neuronal sub population. MSP mRNA is expressed at high levels in the peripheral target fields of DRG somatic afferents, but is undetectable in DRG, spinal cord or freshly dissected sciatic nerve. These results suggest that MSP is a peripheral target derived neurotrophic factor for NGF-responsive adult DRG neurons. PMID- 19285137 TI - Structure-activity relationships of a caged thrombin binding DNA aptamer: insight gained from molecular dynamics simulation studies. AB - 15-mer ssDNA aptamers play a vital role in the inhibition of alpha-thrombin in the blood clotting mechanism. It is of high importance to explore the structural factors controlling the inhibitory nature of the aptamer. Here we investigated the structure-function relationship of the anti-thrombin aptamer, as well as its 'caged' variant (2-(2-nitrophenyl)-propyl group (NPP)) by molecular dynamics simulations. The stability of the unmodified aptamer at different temperatures is examined in 2ns all-atom simulations and compared to experiment. The change in structure when introducing the photo-labile caged compound is analyzed, and the regiospecificity of this modification explained on atomic level. Removal of the photo-labile group leads to the reformation of the active aptamer structure from its inactive state. The mechanism for this formation process is a concerted movement of the aptamer backbone and some highly important bases. The binding of the aptamer to thrombin with regard to the 'caged' group is studied in an explicit simulation with the aptamer-thrombin complex and the reason for the binding/unbinding nature of the aptamer shown. PMID- 19285138 TI - Solution structure of the KdpFABC P-type ATPase from Escherichia coli by electron microscopic single particle analysis. AB - The K+-translocating KdpFABC complex from Escherichia coli functions as a high affinity potassium uptake system and belongs to the superfamily of P-type ATPases, although it exhibits some unique features. It comprises four subunits, and the sites of ATP hydrolysis and substrate transport are located on two different polypeptides. No structural data are so far available for elucidating the correspondingly unique mechanism of coupling ion transport and catalysis in this P-type ATPase. By use of electron microscopy and single particle analysis of negatively stained, solubilized KdpFABC complexes, we solved the structure of the complex at a resolution of 19A, which allowed us to model the arrangement of subunits within the holoenzyme and, thus, to identify the interfaces between subunits. The model showed that the K+-translocating KdpA subunit is in close contact with the transmembrane region of the ATP-hydrolyzing subunit KdpB. The cytosolic C-terminal domain of the KdpC subunit, which is assumed to play a role in cooperative ATP binding together with KdpB, is located in close vicinity to the nucleotide binding domain of KdpB. Overall, the arrangement of subunits agrees with biochemical data and the predictions on subunit interactions. PMID- 19285139 TI - Interpreting single trial data using groupwise regularisation. AB - Univariate statistical approaches are often used for the analysis of neuroimaging data but are unable to detect subtle interactions between different components of brain activity. In contrast, multivariate approaches that use classification as a basis are well-suited to detect such interactions, allowing the analysis of neuroimaging data on the single trial level. However, multivariate approaches typically assign a non-zero contribution to every component, making interpretation of the results troublesome. This paper introduces groupwise regularisation as a novel method for finding sparse, and therefore easy to interpret, models that are able to predict the experimental condition to which single trials belong. Furthermore, the obtained models can be constrained in various ways by placing features extracted from the data that are thought to belong together into groups. In order to learn models from data, we introduce a new algorithm that makes use of stability conditions that have been derived in this paper. The algorithm is used to classify multisensor EEG signals recorded for a motor imagery task using (groupwise) regularised logistic regression as the underlying classifier. We show that regularisation dramatically reduces the number of features without reducing the classification rate. This improves model interpretability as it finds features in the data such as mu and beta desynchronisation in the motor cortex contralateral to the imagined movement. By choosing particular groupings we can constrain the regularised solutions such that a lower number of sensors is used or a model is obtained that generalises well over subjects. The identification of a small number of groups of features that best explain the data make groupwise regularisation a useful new tool for single trial analysis. PMID- 19285140 TI - Electromagnetic responses to invisible face stimuli during binocular suppression. AB - When two incompatible monocular images compete for perceptual dominance, one can dominate awareness while the other is suppressed and invisible. It has been suggested that binocular suppression of this kind involves competitive neuronal interactions at multiple levels of the visual processing hierarchy. Here we asked whether in such situations, complex stimulus information is still preserved at advanced visual processing stages despite competitive interactions at early levels of the visual system. We tested whether face stimuli made invisible through binocular suppression would evoke category-specific responses in the human brain, using magnetoencephalography. During scanning participants were presented with face or house stimuli that were either visible or rendered invisible using continuous flash suppression (CFS). Online behavioural assessment with a two-alternative forced choice task confirmed that participants were neither able to detect target stimuli on CFS trials nor to correctly classify the stimuli as faces or houses. Even though the amplitude of the M170, a well established face-specific signal, was strongly reduced by CFS, there was still a significantly greater signal in response to invisible face compared to invisible house stimuli at the latency and the location of the M170 to visible stimuli. Thus, visual object stimuli undergo category-specific processing in the ventral visual pathway even when profoundly suppressed from awareness by binocular competition. PMID- 19285141 TI - Genetic determinants of target and novelty-related event-related potentials in the auditory oddball response. AB - Processing of novel and target stimuli in the auditory target detection or 'oddball' task encompasses the chronometry of perception, attention and working memory and is reflected in scalp recorded event-related potentials (ERPs). A variety of ERP components related to target and novelty processing have been described and extensively studied, and linked to deficits of cognitive processing. However, little is known about associations of genotypes with ERP endophenotypes. Here we sought to elucidate the genetic underpinnings of auditory oddball ERP components using a novel data analysis technique. A parallel independent component analysis of the electrophysiology and single nucleotide polymorphism (SNP) data was used to extract relations between patterns of ERP components and SNP associations purely based on an analysis incorporating higher order statistics. The method allows for broader associations of genotypes with phenotypes than traditional hypothesis-driven univariate correlational analyses. We show that target detection and processing of novel stimuli are both associated with a shared cluster of genes linked to the adrenergic and dopaminergic pathways. These results provide evidence of genetic influences on normal patterns of ERP generation during auditory target detection and novelty processing at the SNP association level. PMID- 19285142 TI - Left posterior parietal cortex participates in both task preparation and episodic retrieval. AB - Optimal memory retrieval depends not only on the fidelity of stored information, but also on the attentional state of the subject. Factors such as mental preparedness to engage in stimulus processing can facilitate or hinder memory retrieval. The current study used functional magnetic resonance imaging (fMRI) to distinguish preparatory brain activity before episodic and semantic retrieval tasks from activity associated with retrieval itself. A catch-trial imaging paradigm permitted separation of neural responses to preparatory task cues and memory probes. Episodic and semantic task preparation engaged a common set of brain regions, including the bilateral intraparietal sulcus (IPS), left fusiform gyrus (FG), and the pre-supplementary motor area (pre-SMA). In the subsequent retrieval phase, the left IPS was among a set of frontoparietal regions that responded differently to old and new stimuli. In contrast, the right IPS responded to preparatory cues with little modulation during memory retrieval. The findings support a strong left-lateralization of retrieval success effects in left parietal cortex, and further indicate that left IPS performs operations that are common to both task preparation and memory retrieval. Such operations may be related to attentional control, monitoring of stimulus relevance, or retrieval. PMID- 19285143 TI - Visual features of an observed agent do not modulate human brain activity during action observation. AB - Recent neuroimaging evidence in macaques has shown that the neural system underlying the observation of hand actions performed by others (i.e., "action observation system") is modulated by whether the observed action is performed by a person in full view or an isolated hand (i.e., type of view manipulation). Although a human homologue of such circuit has been identified, whether in humans the neural processes involved in this capacity are modulated by the type of view remains unknown. Here we used functional magnetic resonance imaging (fMRI) to investigate whether the "action observation system", with specific reference to the ventral premotor cortex, responds differentially depending on type of view. We also tested this manipulation within regions of the human brain showing overlapping activity for both the observation and the execution of action ("mirror" regions). To this end, the same subjects were requested to observe grasping actions performed under the two types of view (observation conditions) or to perform a grasping action (execution condition). Results from whole-brain analyses indicate that overlapping activity for action observation and execution was evident in a broad network of areas including parietal, premotor and temporal cortices. Activity within such network was evident for both the observation of a person in full view or an isolated hand, but it was not modulated by the type of view. Similarly, results from region of interest (ROI) analyses, performed within the ventral premotor cortex, did confirm that this area responded in a similar fashion following the observation of either an isolated hand or an entire model acting. These findings offer novel insights on what the "action observation" and the "mirror" systems visually code and how the processing underlying such coding may vary across species. Further, they support the hypothesis that action goal is amongst the main determinants for the revelation of action observation activity, and to the existence of a broad system involved in the simulation of action. PMID- 19285144 TI - Against hyperacuity in brain reading: spatial smoothing does not hurt multivariate fMRI analyses? AB - Recently it has been suggested that multivariate analyses of functional magnetic resonance imaging (fMRI) data can detect high spatial frequency components of cortical signals, like sub-millimeter columns. This 'hyperacuity' seems to be at odds with the common assumption that the fMRI signal has a low spatial resolution due to the spatial spread of the underlying hemodynamic events. To resolve this apparent contradiction, I checked a very straightforward prediction of the hyperacuity hypothesis: if multivariate analyses are picking up a small-scale functional organization, then it can be expected that smoothing will be detrimental to the ability to decode these fine-scale spatial signals. I tested this prediction using data obtained with two paradigms to which multivariate techniques have been applied previously, including the decoding of grating orientation from the pattern of activity in primary visual cortex. It was found that smoothing does not decrease the sensitivity of multivariate analyses. Further simulations in which the scale of cortical organization was known indicate that this effect of smoothing contradicts the idea that the patterns detected with multivariate techniques reflect a fine-scale spatial organization. PMID- 19285145 TI - RABBIT: rapid alignment of brains by building intermediate templates. AB - A brain image registration algorithm, referred to as RABBIT, is proposed to achieve fast and accurate image registration with the help of an intermediate template generated by a statistical deformation model. The statistical deformation model is built by principal component analysis (PCA) on a set of training samples of brain deformation fields that warp a selected template image to the individual brain samples. The statistical deformation model is capable of characterizing individual brain deformations by a small number of parameters, which is used to rapidly estimate the brain deformation between the template and a new individual brain image. The estimated deformation is then used to warp the template, thus generating an intermediate template close to the individual brain image. Finally, the shape difference between the intermediate template and the individual brain is estimated by an image registration algorithm, e.g., HAMMER. The overall registration between the template and the individual brain image can be achieved by directly combining the deformation fields that warp the template to the intermediate template, and the intermediate template to the individual brain image. The algorithm has been validated for spatial normalization of both simulated and real magnetic resonance imaging (MRI) brain images. Compared with HAMMER, the experimental results demonstrate that the proposed algorithm can achieve over five times speedup, with similar registration accuracy and statistical power in detecting brain atrophy. PMID- 19285146 TI - Molecular phylogeny of the Drosophila tripunctata and closely related species groups (Diptera: Drosophilidae). AB - We suggest a new phylogenetic hypothesis for the tripunctata radiation based on sequences of mitochondrial genes. Phylogenetic trees were reconstructed by parsimony, maximum likelihood and Bayesian methods. We performed tests for hypotheses of monophyly for taxonomic groups and other specific hypotheses. Results reject the monophyly for the tripunctata group whereas monophyly is not rejected for the tripunctata radiation and other specific groups within the radiation. Although most of the basal nodes were unresolved we were able to identify four clusters within the tripunctata radiation. These results suggest the collection of additional data before a proper taxonomic revision could be proposed. PMID- 19285147 TI - Anti-biofilm properties of Satureja hortensis L. essential oil against periodontal pathogens. AB - Essential oils of several plants are widely used in ethnomedicine for their antimicrobial and anti-inflammatory properties. However, very limited data exist on their use in connection to periodontal diseases. The aim of the present study was to investigate the bacterial growth inhibiting and anti-biofilm effects of Satureja hortensis L. (summer savory), Salvia fruticosa M. (sage), Lavandula stoechas L. (lavender), Myrtus communis L., and Juniperus communis L. (juniper) essential oils. Chemical compositions of the essential oils were analyzed by gas chromatography-mass spectrometry, minimum inhibitor concentrations (MICs) with the agar dilution method, and anti-biofilm effects by the microplate biofilm assay. The toxicity of each essential oil was tested on cultured keratinocytes. Of the 5 essential oils, S. hortensis L. essential oil had the strongest growth inhibition effect. Subinhibitory dose of S. hortensis L. essential oil had anti biofilm effects only against Prevotella nigrescens. Essential oils did not inhibit keratinocyte viability at the concentrations of 1 and 5 microl/ml, however at the concentration of 5 microl/ml epithelial cells detached from the culture well bottom. The present findings suggest that S. hortensis L. essential oil inhibits the growth of periodontal bacteria in the concentration that is safe on keratinocytes, however, in the subinhibitory concentration its anti-biofilm effect is limited. PMID- 19285148 TI - Functional characterization of a C-4 sterol methyl oxidase from the endomycorrhizal fungus Glomus intraradices. AB - Sterols are crucial components of eukaryotic membranes that control membrane fluidity and permeability. They play an important role in cell signaling, polarity and sorting. Since many steps in the pathway are essential, sterol biosynthesis inhibitors (SBI) are widely used as antifungal agents. This work reports the identification and the characterization of a C-4 sterol methyl oxidase (SMO), the first gene involved in the sterol biosynthetic pathway, so far described from an arbuscular mycorrhizal fungus. The sequence, called GintSMO, shows a primary structure, a hydrophobicity profile and a pattern of histidine rich motifs which are typical of C-4 methyl sterol oxidases. The complementation assay in a Saccharomyces cerevisiae mutant strain demonstrates that GintSMO encodes a functional SMO. Changes in GintSMO transcript levels and in the amount of the sterol precursor squalene were observed in in vitro grown extraradical structures exposed to the fenpropimorph SBI fungicide. PMID- 19285149 TI - Biosynthesis of the cyclooligomer depsipeptide bassianolide, an insecticidal virulence factor of Beauveria bassiana. AB - Beauveria bassiana is a facultative entomopathogen with an extremely broad host range that is used as a commercial biopesticide for the control of insects of agricultural, veterinary and medical significance. B. bassiana produces bassianolide, a cyclooligomer depsipeptide secondary metabolite. We have cloned the bbBsls gene of B. bassiana encoding a nonribosomal peptide synthetase (NRPS). Targeted inactivation of the B. bassiana genomic copy of bbBsls abolished bassianolide production, but did not affect the biosynthesis of beauvericin, another cyclodepsipeptide produced by the strain. Comparative sequence analysis of the BbBSLS bassianolide synthetase revealed enzymatic domains for the iterative synthesis of an enzyme-bound dipeptidol monomer intermediate from d-2 hydroxyisovalerate and l-leucine. Further BbBSLS domains are predicted to catalyze the formation of the cyclic tetrameric ester bassianolide by recursive condensations of this monomer. Comparative infection assays against three selected insect hosts established bassianolide as a highly significant virulence factor of B. bassiana. PMID- 19285150 TI - Gas exchange in avian embryos and hatchlings. AB - The avian egg has been proven to be an excellent model for the study of the physical principles and the physiological characteristics of embryonic gas exchange. In recent years, it has become a model for the studies of the prenatal development of pulmonary ventilation, its chemical control and its interaction with extra-pulmonary gas exchange. Differently from mammals, in birds the initiation of pulmonary ventilation and the transition from diffusive to convective gas exchange are gradual and slow-occurring events amenable to detailed investigations. The absence of the placenta and of the mother permits the study of the mechanisms of embryonic adaptation to prenatal perturbations in a way that would be impossible with mammalian preparations. First, this review summarises the general aspects of the natural history of the avian egg that are pertinent to embryonic metabolism, growth and gas exchange and the characteristics of the structures participating in gas exchange. Then, the review focuses on the embryonic development of pulmonary ventilation, its regulation in relation to the embryo's environment and metabolic state, the effects that acute or sustained changes in embryonic temperature or oxygenation can have on growth, metabolism and ventilatory control. PMID- 19285151 TI - Effects of point mutations on the structural stability of tuna myoglobins. AB - Structural stabilities of myoglobin (Mb) from several tuna fish species significantly differ from each other, although the amino acid sequence identities are very high (>95%), suggesting that limited number of substitutions greatly affect the stability of Mb. To address this hypothesis, attempts were made to elaborate recombinant tuna Mbs with point mutations on the different residues among fish Mbs. The expression plasmid constructs were based on bigeye tuna Mb cDNA sequence, and the recombinant proteins were expressed as GST-fusion proteins in Escherichia coli. After removal of the GST segment and affinity purification, the stability of five Mb mutants, namely, A49G, T91K, K92Q, V108A, and H112Q, together with the wild type (WT) were measured, taking temperature dependency of alpha-helical content and denaturant (urea and guanidine-HCl) concentration dependency of Soret band absorbance as parameters. As a result, the mutant H112Q showed much higher stability than WT, while the structures of K92Q, T91K and A49G mutants were destabilized. No essential change in helical content was observed for V108A, but the mutant was found to be destabilized easier by the denaturants. These findings suggested that the highly conserved residues among tuna species are responsible for their stability of Mbs, but a few non-conserved residues dramatically give rise to the differences in stability of Mbs among species. PMID- 19285152 TI - Prevalence of Capnocytophaga canimorsus in dogs and occurrence of potential virulence factors. AB - Capnocytophaga canimorsus is a Gram-negative commensal of dog's mouth causing severe human infections. A strain isolated from a human fatal infection was recently shown to have a sialidase, to inhibit the bactericidal activity of macrophages and to block the release of nitric oxide by LPS-stimulated macrophages. The present study aimed at determining the prevalence of C. canimorsus in dogs and the occurrence of these hypothetical virulence factors. C. canimorsus could be retrieved from the saliva of 61 dogs out of 106 sampled. Like in clinical isolates, all dog strains had a sialidase and 60% blocked the killing of phagocytosed Escherichia coli by macrophages. In contrast, only 6.5% of dog strains blocked the release of nitric oxide by LPS-challenged macrophages, suggesting that this property might contribute to virulence. The comparative analysis of 69 16S rDNA sequences revealed the existence of C. canimorsus strains that could be misdiagnosed. PMID- 19285153 TI - The influence of temporal lobectomy on the mental state of patients with psychomotor epilepsy. T. Alajouanine, J. Nehlil, R. Houdart. Revue Neurol 1958;98(3):165-171. 1958. PMID- 19285154 TI - Use of a commercial line blot assay as a screening test for autoantibodies in inflammatory myopathies. AB - AIMS: To evaluate the clinical utility of a commercial immunoblot assay for the detection of myositis-specific autoantibodies. METHODS: Serum samples from 153 myositis patients and 77 disease controls were investigated. The commercial Euroline assay with seven autoantigens (Mi-2, Ku, PM-Scl, Jo-1, Pl-7, Pl-12 and SSA/Ro-52) was used according to the manufacturer s instructions, and supplemented with an anti-SRP strip. In a separate experiment analyses were performed at different temperatures. Results were recorded with densitometry. RESULTS: Anti-Jo-1 was found in 18 myositis and one systemic sclerosis patient. Antibodies against Mi-2 were found in 5 myositis patients, and eleven myositis patients had antibodies against PM-Scl. Four myositis patients showed anti-Pl-7 reactivity, whereas no patients had antibodies against Pl-12. Anti-Ku antibodies were found in 4 myositis and 2 primary Sjogren's syndrome patients. Anti-SRP was found in 8 myositis patients as well as in two disease controls. Antibodies against SSA/Ro52 ranged between 23-62% in all groups except juvenile dermatomyositis patients. Most autoantibody reactivities were clearly positive, only 11% (14/127) were borderline positive. Higher assay temperature increased antibody reactivities. CONCLUSIONS: Except for anti-SSA/Ro-52 and anti-Ku the antibody reactivities were rather myositis-specific, supporting the use of this immunoblot assay. However, assay validation needs to be determined against other methods. PMID- 19285155 TI - The therapeutic potential of targeting B cells and anti-oxLDL antibodies in atherosclerosis. AB - While the involvement of T cells in atherosclerosis is nowadays well accepted, little is known about the role of B cells. Obviously, B cells as the source of antibodies, in particular antibodies to oxLDL, have gained a lot of attention in atherosclerosis. In addition, B cells do harbour other functions in adaptive immunity. In this review, we provide an overview of the current knowledge on both the role of B cells and antibodies, i.e., anti-oxLDL antibodies, in atherosclerosis. It appears that B cells and also anti-oxLDL antibodies may comprise pro- and anti-atherogenic effects. Therefore, the establishment of effective therapy, targeting B cells or anti-oxLDL antibodies, warrants further research to unravel these opposite effects. PMID- 19285156 TI - IL-18 and skin inflammation. AB - IL-18 belongs to the IL-1 family of cytokines and has recently regained interest in the context of inflammasome activation. The inflammasome dependent caspase 1 cleaves pro-IL-18 into the active form - similar to what is known for IL-1ss. Still, the action and importance of IL-18 are not completely understood. There are several indications that it plays a pathogenetically important role in chronic inflammatory conditions of epithelial organs (such as skin, gut, kidney) and importantly also in responses against self. Here, we summarise current knowledge on the role of IL-18 in human skin inflammation with a focus on its role in Cutaneous Lupus Erythematosus (CLE). There is evidence that IL-18 plays a role in CLE upstream of TNFalpha. In CLE but not normal keratinocytes IL-18 strongly induces TNFalpha release, which then results in apoptosis. Blocking TNFalpha in vitro prevents apoptosis of keratinocytes but anti-TNFalpha therapy is not applicable in LE conditions. We will discuss potential approaches to control IL-18 in skin inflammation. PMID- 19285157 TI - Utilization of synthetic peptides to evaluate the importance of substrate interaction at the proteolytic site of Escherichia coli Lon protease. AB - Lon, also known as protease La, is an ATP-dependent protease functioning to degrade many unstructured proteins. Currently, very little is known about the substrate determinants of Lon at the proteolytic site. Using synthetic peptides constituting different regions of the endogenous protein substrate lambdaN, we demonstrated that the proteolytic site of Escherichia coli Lon exhibits a certain level of localized sequence specificity. Using an alanine positional scanning approach, we discovered a set of discontinuous substrate determinants surrounding the scissile Lon cleavage site in a model peptide substrate, which function to influence the k(cat) of the peptidase activity of Lon. We further investigated the mode of peptide interaction with the proteolytically inactive Lon mutant S679A in the absence and presence of ADP or AMPPNP by 2-dimensional nuclear magnetic resonance spectroscopy, and discovered that the binding interaction between protein and peptide varies with the nucleotide bound to the enzyme. This observation is suggestive of a substrate translocation step, which likely limits the turnover of the proteolytic reaction. The contribution of the identified substrate determinants towards the kinetics of ATP-dependent degradation of lambdaN and truncated lambdaN mutants by Lon was also examined. Our results indicated that Lon likely recognizes numerous discontinuous substrate determinants throughout lambdaN to achieve substrate promiscuity. PMID- 19285158 TI - A synonymous polymorphism in a common MDR1 (ABCB1) haplotype shapes protein function. AB - The MDR1 (ABCB1) gene encodes a membrane-bound transporter that actively effluxes a wide range of compounds from cells. The overexpression of MDR1 by multidrug resistant cancer cells is a serious impediment to chemotherapy. MDR1 is expressed in various tissues to protect them from the adverse effect of toxins. The pharmacokinetics of drugs that are also MDR1 substrates also influence disease outcome and treatment efficacy. Although MDR1 is a well-conserved gene, there is increasing evidence that its polymorphisms affect substrate specificity. Three single nucleotide polymorphisms (SNPs) occur frequently and have strong linkage, creating a common haplotype at positions 1236C>T (G412G), 2677G>T (A893S) and 3435C>T (I1145I). The frequency of the synonymous 3435C>T polymorphism has been shown to vary significantly according to ethnicity. Existing literature suggests that the haplotype plays a role in response to drugs and disease susceptibility. This review summarizes recent findings on the 3435C>T polymorphism of MDR1 and the haplotype to which it belongs. A possible molecular mechanism of action by ribosome stalling that can change protein structure and function by altering protein folding is discussed. PMID- 19285159 TI - Identification of polyubiquitin binding proteins involved in NF-kappaB signaling using protein arrays. AB - Attachment of ubiquitin to proteins represents a central mechanism for the regulation of protein metabolism and function. In the NF-kappaB pathway, binding of NEMO to polyubiquitinated substrates initiates the pathway in response to cellular stimuli. Other polyubiquitin binding proteins can antagonize this pathway by competing with NEMO for polyubiquitin. We have used protein arrays to identify polyubiquitin binding proteins that regulate NF-kappaB activity. Using polyubiquitin as bait, protein arrays were screened and polyubiquitin binders identified. Novel polyubiquitin binders AWP1, CALCOCO2, N4BP1, RIO3, TEX27, TTC3, UBFD1 and ZNF313 were identified using this approach, while known NF-kappaB regulators including NEMO, A20, ABIN-1, ABIN-2, optineurin and p62 were also identified. Overexpressed AWP1 and RIO3 repressed NF-kappaB activity in a manner similar to optineurin, while siRNAs directed against AWP1 and RIO3 also reduced NF-kappaB activity. TNFalpha-dependent degradation of IkappaBalpha was also suppressed by overexpression of AWP1 and RIO3, possibly due to the polyubiquitin binding activity of these proteins. Protein array screening using polyubiquitin enabled rapid identification of many known and novel polyubiquitin binding proteins and the identification of novel NF-kappaB regulators. PMID- 19285160 TI - An easy way of generating operative schematic diagrams. PMID- 19285162 TI - Slow down, youth's food's too fast. PMID- 19285161 TI - Human osteoarthritic chondrocytes are impaired in matrix metalloproteinase-13 inhibition by IFN-gamma due to reduced IFN-gamma receptor levels. AB - OBJECTIVE: Human osteoarthritic (OA) cartilage type-II collagen is preferentially cleaved by the proinflammatory cytokine-induced matrix metalloproteinases-13 (MMP 13). Interferon-gamma (IFN-gamma) potently inhibits interleukin-1 (IL-1)-induced MMP-13 expression in healthy chondrocytes. Our goal was to study the previously unknown impact of IFN-gamma on MMP-13 in OA and compare the levels and functional activity of IFN-gamma receptor (IFN-gammaR1) in healthy and OA chondrocytes. METHODS: Chondrocytes were obtained from OA patients and non-arthritic control subjects and treated with IL-1+ or- IFN-gamma. MMP-13 mRNA and protein expression were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. IFN-gammaR1 expression was assessed by flow cytometry, immunoprecipitation and immunohistochemistry with fluorescein-labeled antibody. IFN-gammaR1 was neutralized with its antibody and signal transducer and activator of transcription 1 (STAT1) phosphorylation analyzed by Western blotting. OA chondrocytes were also transfected with control and IFN-gammaR1 expression vectors. RESULTS: OA chondrocytes displayed a drastically impaired MMP-13 suppression by IFN-gamma compared to control cells. IFN-gammaR1 levels were significantly decreased in OA chondrocytes as assessed by flow cytometry, immunoprecipitation and immunohistochemistry. Consequently, IFN-gamma-stimulated STAT1 phosphorylation mediated by IFN-gammaR1 was also considerably reduced in OA patient chondrocytes. IFN-gammaR1 overexpression in OA cells restored MMP-13 suppression by IFN-gamma. CONCLUSIONS: Ability of IFN-gamma to suppress IL-1 induced MMP-13 expression is diminished in OA chondrocytes due to decreased IFN gammaR1 levels, activity and impaired downstream signal transduction. Therefore, IFN-gammaR1 modulation and weakened endogenous IFN-gamma response may be important mechanisms in OA pathogenesis and cartilage degradation. PMID- 19285163 TI - Clinical policy: neuroimaging and decisionmaking in adult mild traumatic brain injury in the acute setting. AB - This clinical policy provides evidence-based recommendations on select issues in the management of adult patients with mild traumatic brain injury (TBI) in the acute setting. It is the result of joint efforts between the American College of Emergency Physicians and the Centers for Disease Control and Prevention and was developed by a multidisciplinary panel. The critical questions addressed in this clinical policy are: (1) Which patients with mild TBI should have a noncontrast head computed tomography (CT) scan in the emergency department (ED)? (2) Is there a role for head magnetic resonance imaging over noncontrast CT in the ED evaluation of a patient with acute mild TBI? (3) In patients with mild TBI, are brain specific serum biomarkers predictive of an acute traumatic intracranial injury? (4) Can a patient with an isolated mild TBI and a normal neurologic evaluation result be safely discharged from the ED if a noncontrast head CT scan shows no evidence of intracranial injury? Inclusion criteria for application of this clinical policy's recommendations are nonpenetrating trauma to the head, presentation to the ED within 24 hours of injury, a Glasgow Coma Scale score of 14 or 15 on initial evaluation in the ED, and aged 16 years or greater. The primary outcome measure for questions 1, 2, and 3 is the presence of an acute intracranial injury on noncontrast head CT scan; the primary outcome measure for question 4 is the occurrence of neurologic deterioration. PMID- 19285165 TI - "I wanted to come in early, before it got busy.". PMID- 19285166 TI - New graduate nurses in the emergency department. PMID- 19285167 TI - Graduate nurses in the emergency department. PMID- 19285168 TI - ED patient falls and resulting injuries. AB - INTRODUCTION: Patient falls are the most common adverse events reported in hospitals. There is a growing body of literature on inpatient falls but a lack of data on ED falls. We applied the Hendrich II Fall Risk Model to patients who fell during their ED stays and provided a description of the patients and their injuries. METHODS: We retrospectively reviewed the medical records of all patients who fell in the emergency department during a 2-year period. We collected the 8 assessment parameters for high-risk fall identification in the Hendrich II Fall Risk Model. We also collected subject characteristics, circumstances surrounding the falls, fall-related injuries, and ED disposition. RESULTS: Fifty-seven falls were recorded, representing a rate of 0.288 falls per 1000 patient visits. The average age was 50 years, and a median of 48. 67% were men. Twenty-one subjects had a Hendrich II Model score of 5 of greater, which represents a sensitivity of 37.5%. Eleven subjects (19.6%) were intoxicated with alcohol. Eleven subjects (19.6%) received a potentially sedating medication prior to the fall. Thirty-six subjects (64.3%) fell in their ED rooms. Six subjects (10.7%) fell in the restroom. Three falls (5.4%) resulted in lacerations and 2 falls (3.6%) resulted in hematomas. DISCUSSION: The Hendrich II Fall Risk Model may not reliably identify patients at high risk of falling in the ED setting. It may be necessary to develop an emergency department-specific fall model considering additional factors, such as intoxication and receipt of potentially sedating medications. PMID- 19285169 TI - Blood glucose measurement in patients with suspected diabetic ketoacidosis: a comparison of Abbott MediSense PCx point-of-care meter values to reference laboratory values. AB - INTRODUCTION: The purpose of this study is to compare blood glucose levels measured by a point of care (POC) device to laboratory measurement using the same sample venous blood from patients with suspected diabetic ketoacidosis (DKA). METHODS: A descriptive correlational design was used for this IRB-approved quality assurance project. The study site was the 50-bed BMC emergency department (ED) which has an annual census of over 100,000 patient visits. The convenience sample consisted of 54 blood samples from suspected DKA patients with orders for hourly blood draws for glucose measurement. Spearman correlations of the glucose POC values, reference lab values, and differences between the two, were evaluated. A chi-square test was used to evaluate the association between the acidosis status and FDA acceptability of POC values. RESULTS: Patient age range was 10-86 years; 63% were females; 46% had a final diagnosis of DKA. POC values underestimated glucose levels 93% of the time. There was a high correlation between the lab value and the magnitude of the difference, (lab minus POC value) indicating that the higher the true glucose value, the greater the difference between the lab and the POC value. A chi-square test showed no overall association between acidosis and FDA-acceptability. DISCUSSION: The POC values underestimated lab reported glucose levels in 50 of 54 cases even with the use of same venous sample sent to the lab, which make it highly unreliable for use in monitoring suspected DKA patients. PMID- 19285170 TI - Family violence intervention within an emergency department: achieving change requires multifaceted processes to maximize safety. AB - INTRODUCTION: Family violence is common with significant long-term negative health effects. Health professionals are recognised as key providers of family violence intervention. In 2002, the Hawke's Bay District Health Board launched a Family Violence Intervention Programme in its emergency department. The intervention programme involved staff training, the development of resources and routine questioning for partner abuse within the social history for all women 16 years and over. The aim was to identify the barriers and enablers to routine questioning one year after the programme was launched to inform programme improvements. METHODS: Evaluation research using semi-structured interviews; eleven staff participated in either a single or a group interview. Content and thematic analysis, with triangulation of findings was used. RESULTS: The interviews revealed that routine questioning for partner abuse is difficult in the emergency department. Some staff screened routinely while others only offered intervention when overt abuse was identified. Barriers, enablers and solutions revealed by participants were either personal or organisational; all had the common theme of safety. DISCUSSION: Routine questioning for partner abuse is challenging and its introduction into practice requires a systems approach to achieve change. Barriers to questioning exist and by simultaneously addressing these and implementing enablers, at an organisational and personal level, barriers are eliminated or at least minimised. A link was evident between nurses' level of comfort and their rate of questioning. A multifaceted approach focusing on safety of all concerned can support change resulting in implementation of family violence intervention in the health sector. PMID- 19285171 TI - Patients who leave the emergency department without being seen. AB - INTRODUCTION: Patients who present to the ED for care and leave without being seen (LWBS) represent a significant problem. The objective of this study was to determine why patients LWBS, how long they perceived waiting versus actual time waited before leaving, and factors that might have prevented LWBS. METHODS: We conducted a prospective, scripted phone survey of all patients who left without being seen over a two-month period in 2006 at an ED with approximately 65,000 yearly visits. Outcome measures were number leaving, ability to obtain care after leaving, reason for leaving, would they return to this ED, perceived and actual time waited, number with a primary physician, and factors associated with leaving. RESULTS: One-hundred and twenty-seven of 11,147 total patients (1.1%) patients left without being seen. Seventy-two (56.7%) were interviewed within 8 days. Eighty-four and seven-tenths percent stated they had a primary physician. The mean age was 29.9 years, and 44.4% were male. The patient-reported mean time waited before leaving was 73.2 minutes while the actual mean time waited was 70.4 minutes. The reasons for leaving were the length of wait (76.7%), the problem resolved (12.3%), and for other reasons (11.0%). During the week after leaving the ED, 56.3% were able to obtain medical care. Sixty-five percent would seek future emergency care at this ED, 15.3% would not, and 19.7% would possibly return. During the wait, patients wanted information, lab tests/X-rays, and analgesics. DISCUSSION: Most would return for future ED care. Most had a physician and were able to obtain care elsewhere. Reduced LWBS might be accomplished by triage testing, communication and attention to pain. PMID- 19285172 TI - Using medical screening examinations to reduce emergency department overcrowding. PMID- 19285173 TI - The use of negative inspiratory force by ED personnel to monitor respiratory deterioration in the event of a botulism-induced MCI. PMID- 19285174 TI - Reducing time to first cardiac marker results by integrating prehospital and ED protocols. PMID- 19285175 TI - ECG interpretation part 2: determination of bundle branch and fascicular blocks. PMID- 19285176 TI - Illegal immigrants in the emergency department: an ethical dilemma for nurses? PMID- 19285177 TI - Caring for the older person in the emergency department: the ASET program and the role of the ASET clinical nurse consultant in South Western Sydney, Australia. PMID- 19285178 TI - A 15-year-old trauma patient with lower extremity paralysis. PMID- 19285180 TI - Discharge vital signs: an enhancement to ED quality and patient outcomes. PMID- 19285181 TI - Coaching for success: sustaining change in emergency care. PMID- 19285182 TI - Drifting into "at-risk" behaviors: is your emergency department in jeopardy? PMID- 19285183 TI - The power of nurses in the legislative process. PMID- 19285184 TI - The role of collaborative Web publishing tools in evidence-based practice. PMID- 19285185 TI - Flooding in Cedar Rapids, Iowa. PMID- 19285186 TI - Helicopter air medical transport safety: what is the role of the emergency nurse? PMID- 19285187 TI - Emergency nursing in Lebanon amidst chaos. PMID- 19285188 TI - Cutting-edge discussions of management, policy, and program issues in emergency care. PMID- 19285189 TI - A 24-day-old child with projectile vomiting. PMID- 19285190 TI - Rapid dosing of critical care infusions: the dopamine and norepinephrine "clocks". PMID- 19285191 TI - Ehlers-Danlos syndrome in trauma: a case review. PMID- 19285192 TI - Death at the front door: lessons learned when a bomb threatens triage. PMID- 19285193 TI - Functional genome research on bacteria relevant for agriculture, environment and biotechnology. PMID- 19285194 TI - Challenging the notion of an early-onset of cognitive decline. AB - Salthouse claims that cognitive aging starts around 20 years of age. The basis for this claim is cross-sectional data. He dismisses longitudinal data, which typically show the cognitive decline to start much later, around 60 years of age. He states that longitudinal data cannot be trusted because they are flawed. There is a confounding between the effects of maturation and retest effects. We challenge Salthouse's strong claim on four accounts. PMID- 19285195 TI - Substance use and delinquency among fifth graders who have jobs. AB - BACKGROUND: Working for pay is associated with substance use and delinquency among older adolescents, although information is scant about younger youth who work. This study investigates associations between self-reports of having a job and substance use and delinquent behaviors in a sample of U.S. 5th graders. METHODS: A total of 5147 5th graders and their parents from three large metropolitan areas were assessed in a cross-sectional survey between Fall 2004 and Summer 2006. Multivariate regression was used to estimate associations between having a job and substance use and delinquency. Analyses were conducted in Fall 2007. RESULTS: Twenty-one percent of 5th graders reported having a job, with most working <5 hours per week. Typical jobs included yard work, babysitting, and cleaning. In multivariate models that controlled for demographic characteristics, household composition, and household income, having a job was significantly associated with past-30-day use of tobacco (OR=2.2), alcohol (OR=1.7), and marijuana (OR=3.1). Having a job was also significantly associated with ever being in a fight (OR=1.5) and with running away from home (OR=1.8). Further analyses indicated that the associations between holding a job and delinquency outcomes were driven largely by young workers who worked >2 hours per week. CONCLUSIONS: Among 5th graders, having a job was associated with substance using behaviors and delinquency. Clinicians should consider asking young patients whether they work, and stress to parents the importance of monitoring the work activities, workplaces, and associates of their children. PMID- 19285196 TI - Exposure to sexual lyrics and sexual experience among urban adolescents. AB - BACKGROUND: Two thirds of all sexual references in music are degrading in nature, yet it remains uncertain whether these references promote earlier sexual activity. The purpose of this study was to determine if exposure to lyrics describing degrading sex in popular music is independently associated with sexual behavior in a cohort of urban adolescents. METHODS: All ninth-grade health students at three large urban high schools completed in-school surveys in 2006 and 2007. Participants' exposure to lyrics describing degrading sex was computed with overall music exposure and content analyses of their favorite artists' songs. Outcomes included sexual intercourse and progression along a noncoital sexual continuum. Multivariable regression was used to assess independent associations between exposure to lyrics describing degrading sex and outcomes. RESULTS: The 711 participants were exposed to 14.7 hours each week of songs with lyrics describing degrading sex (SD=17.0). Almost one third of participants (n=216) had previously been sexually active. Compared to those with the least exposure to lyrics describing degrading sex, those with the most exposure were more than twice as likely to have had sexual intercourse (OR=2.07; 95% CI=1.26, 3.41), even after adjusting for all covariates. Similarly, among those who had not had sexual intercourse, those in the highest tertile of exposure to lyrics describing degrading sex were nearly twice as likely to have progressed along a noncoital sexual continuum (OR=1.88; 95% CI=1.23, 2.88) compared to those in the lowest tertile. Finally, the relationships between exposure to lyrics describing nondegrading sex and sexual outcomes were not significant. CONCLUSIONS: This study supports an association between exposure to lyrics describing degrading sex in popular music and early sexual experience among adolescents. PMID- 19285197 TI - Literacy, numeracy, and portion-size estimation skills. AB - BACKGROUND: Portion-size estimation is an important component of weight management. Literacy and numeracy skills may be important for accurate portion size estimation. It was hypothesized that low literacy and numeracy would be associated with decreased accuracy in portion estimation. METHODS: A cross sectional study of primary care patients was performed from July 2006 to August 2007; analyses were performed from January 2008 to October 2008. Literacy and numeracy were assessed with validated measures (the Rapid Estimate of Adult Literacy in Medicine and the Wide Range Achievement Test, third edition). For three solid-food items and one liquid item, participants were asked to serve both a single serving and a specified weight or volume amount representing a single serving. Portion-size estimation was considered accurate if it fell within +/-25% of a single standard serving. RESULTS: Of 164 participants, 71% were women, 64% were white, and mean (SD) BMI was 30.6 (8.3) kg/m(2). While 91% reported completing high school, 24% had <9th-grade literacy skills and 67% had <9th-grade numeracy skills. When all items were combined, 65% of participants were accurate when asked to serve a single serving, and 62% were accurate when asked to serve a specified amount. In unadjusted analyses, both literacy and numeracy were associated with inaccurate estimation. In multivariate analyses, only lower literacy was associated with inaccuracy in serving a single serving (OR=2.54; 95% CI=1.11, 5.81). CONCLUSIONS: In this study, many participants had poor portion size estimation skills. Lower literacy skills were associated with less accuracy when participants were asked to serve a single serving. Opportunities may exist to improve portion-size estimation by addressing literacy. PMID- 19285198 TI - Capacity for colorectal cancer screening by colonoscopy, Montana, 2008. AB - BACKGROUND: Colorectal cancer is largely preventable by screening, but screening participation is low in Montana. Colonoscopy is often considered the most accurate screening test and has the potential to prevent colon cancer by pre emptive removal of polyps. However, colonoscopy may not be equally available to all residents of rural states. The Montana Department of Public Health and Human Services (DPHHS) has assigned high priority to colorectal cancer prevention, but before beginning a campaign to increase screening, DPHHS conducted a survey to determine existing colonoscopy screening capacity. METHODS: An eight-question survey was sent by DPHHS to all hospitals and ambulatory surgical centers that perform colonoscopy in Montana, assessing their current and projected capacity to perform screening colonoscopies. Data were collected from March to May 2008, and analysis was performed in June 2008. RESULTS: Responses were received from 43 of 44 hospitals and ambulatory surgical centers performing colonoscopies in Montana. The number of screening colonoscopies performed was estimated to be 19,444 per year. Unused colonoscopy screening capacity was estimated to be 23,096 procedures per year. Although similar total capacity existed in urban and rural areas, more unused capacity existed in rural areas. CONCLUSIONS: Montana has statewide capacity to meet moderately increased demand for screening colonoscopy but would be able to meet only 17% of demand in 2009 if all eligible adults chose colonoscopy as their primary form of screening. It is feasible to develop campaigns to increase screening colonoscopy participation now, but a systematic combination of colonoscopy and other screening modalities may be better able to meet Montana's long-term needs. PMID- 19285199 TI - Obesity among those with mental disorders: a National Institute of Mental Health meeting report. AB - The National Institute of Mental Health convened a meeting in October 2005 to review the literature on obesity, nutrition, and physical activity among those with mental disorders. The findings of this meeting and subsequent update of the literature review are summarized here. Levels of obesity are higher in those with schizophrenia and depression, as is mortality from obesity-related conditions such as coronary heart disease. Medication side effects, particularly the metabolic side effects of antipsychotic medications, contribute to the high levels of obesity in those with schizophrenia, but increased obesity and visceral adiposity have been found in some but not all samples of drug-naive patients as well. Many of the weight-management strategies used in the general population may be applicable to those with mental disorders, but little is known about the effects of these strategies on this patient population or how these strategies may need to be adapted for the unique needs of those with mental disorders. The minimal research on weight-management programs for those with mental disorders indicates that meaningful changes in dietary intake and physical activity are possible. Physical activity is an important component of any weight-management program, particularly for those with depression, for which a substantial body of research indicates both mental and physical health benefits. Obesity among those with mental disorders has not received adequate research attention, and empirically-based interventions to address the increasing prevalence of obesity and risk of cardiovascular and metabolic diseases in this population are lacking. PMID- 19285200 TI - Screening for osteoporosis in the adult U.S. population: ACPM position statement on preventive practice. AB - CONTEXT: Osteoporosis is a common and costly disease that is associated with high morbidity and mortality. There is a lack of direct evidence supporting the benefits of bone mineral density (BMD) screening on osteoporosis outcomes. However, there is indirect evidence to support screening for osteoporosis given the availability of medications with good antifracture efficacy. This paper addresses the position of the American College of Preventive Medicine (ACPM) on osteoporosis screening. EVIDENCE ACQUISITION: The medical literature was reviewed for studies examining the benefits and harms of osteoporosis screening. An overview is also provided of available modalities for osteoporosis screening, risk-assessment tools, cost effectiveness, benefits and harms of screening, rationale for the study, and recommendations from leading health organizations and ACPM. A review was done of English language articles published prior to September 2008 that were retrieved via search on PubMed, from references from pertinent review or landmark articles, and from websites of leading health organizations. EVIDENCE SYNTHESIS: There were no randomized controlled trials (RCTs) of osteoporosis screening on fracture outcomes. However, there was one observational study that demonstrated reduced fracture incidence among recipients of BMD testing. Dual energy x-ray absorptiometry is currently one of the most widely accepted and utilized methods for assessing BMD. Other potential tests for detecting osteoporosis include quantitative ultrasound, quantitative computer tomography, and biochemical markers of bone turnover. Testing via BMD is a cost effective method for detecting osteoporosis in both men and women. Osteoporosis risk-assessment tools such as the WHO fracture-risk algorithm are useful supplements to BMD assessments as they provide estimates of absolute fracture risks. They can also be used with or without BMD testing to assist healthcare providers and patients in making decisions regarding osteoporosis treatments. CONCLUSIONS: All adult patients aged >or=50 years should be evaluated for risk factors for osteoporosis. Screening with BMD testing for osteoporosis is recommended in women aged >or=65 years and in men aged >or=70 years. Younger postmenopausal women and men aged 50-69 years should undergo screening if they have at least one major or two minor risk factors for osteoporosis. It is also recommended that clinicians consider using an osteoporosis risk-assessment tool to evaluate absolute fracture risk to determine appropriate osteoporosis therapies. PMID- 19285201 TI - A counterview on data quality and the systematic review process for occupational injury interventions: are we missing the forest for the trees? PMID- 19285203 TI - Measures of the food environment: a compilation of the literature, 1990-2007. AB - BACKGROUND: Valid and reliable measures are required to assess any effect of the food environment on individual dietary behavior, and form the foundation of research that may inform obesity-related policy. Although many methods of measuring the food environment exist, this area of research is still relatively new and there has been no systematic attempt to gather these measures, to compare and contrast them, or to report on their psychometric properties. EVIDENCE ACQUISITION: A structured literature search was conducted to identify peer reviewed articles published between January 1990 and August 2007 that measured the community-level food environment. These articles were categorized into the following environments: food stores, restaurants, schools, and worksites. The measurement strategies in these studies were categorized as instruments (checklists, market baskets, inventories, or interviews/questionnaires) or methodologies (geographic, sales, menu, or nutrient analyses). EVIDENCE SYNTHESIS: A total of 137 articles were identified that included measures of the food environment. Researchers focused on assessing the accessibility, availability, affordability, and quality of the food environment. The most frequently used measure overall was some form of geographic analysis. Eighteen of the 137 articles (13.1%) tested for any psychometric properties, including inter rater reliability, test-retest reliability, and/or validity. CONCLUSIONS: A greater focus on testing for reliability and validity of measures of the food environment may increase rigor in research in this area. Robust measures of the food environment may strengthen research on the effects of the community-level food environment on individual dietary behavior, assist in the development and evaluation of interventions, and inform policymaking targeted at reducing the prevalence of obesity and improving diet. PMID- 19285204 TI - Measuring the food environment: state of the science. AB - The past decades have seen an increased interest in understanding how the environment affects population health. In particular, public health practitioners and researchers alike are eager to know how the food environments of neighborhoods, schools, and worksites affect food choices and, ultimately, population risk for obesity and other diet-related chronic disease. However, the measurement tools for assessing the environment and the employed study designs have limited our ability to gain important ground. The field has not yet fully considered the psychometric properties of the environmental measurement tools, or how to deal with the copious amounts of data generated from many environmental measures. The field is dominated by research using unsophisticated study designs and has frequently failed to see the role of social and individual factors and how they interrelate with the physical environment. This paper examines some of the measurement issues to be considered as public health practitioners and researchers attempt to understand the impact of the food environment on the health of communities and takes a broad look at where the science currently is with regard to how the food environment is measured, thoughts on what issues may benefit from more deliberate inspection, and directions for future work. PMID- 19285205 TI - Measuring food availability and access in African-American communities: implications for intervention and policy. AB - Obesity is a major public health concern in the U.S. As compared to whites, minority populations are disproportionately at risk, with the highest prevalence rates of overweight and obesity occurring among African American women. Although researchers and policymakers argue that environmental approaches have the greatest potential to reverse the rising prevalence of obesity, critical gaps remain in our understanding of the complex mechanisms that underlie the associations between neighborhood food environments and weight status. A major challenge has been the need for reliable and valid measures to assess aspects of the neighborhood food environment that encourage or inhibit healthful eating behaviors and weight management. Investigators have made considerable gains in the development of tools and approaches to measure neighborhood food environments overall, but few studies focus on the specific challenges and issues associated with characterizing neighborhood food environments in communities of color. This paper highlights important considerations for measuring food environments in African-American neighborhoods and their implications for developing programmatic and policy solutions to reduce racial disparities in overweight. PMID- 19285206 TI - Measuring potential access to food stores and food-service places in rural areas in the U.S. AB - Geographic access to healthy food resources remains a major focus of research that examines the contribution of the built environment to healthful eating. Methods used to define and measure spatial accessibility can significantly affect the results. Considering the implications for marketing, policy, and programs, adequate measurement of the food environment is important. Little of the published work on food access has focused on rural areas, where the burden of nutrition-related disease is greater. This article seeks to expand our understanding of the challenges to measurement of potential spatial access to food resources in rural areas in the U.S. Key challenges to the accurate measurement of the food environment in rural areas include: (1) defining the rural food environment while recognizing that market factors may be changing; (2) describing characteristics that may differentiate similar types of food stores and food-service places; and (3) determining location coordinates for food stores and food-service places. In order to enhance measurements in rural areas, "ground truthed" methodology, which includes on-site observation and collection of GPS data, should become the standard for rural areas. Measurement must also recognize the emergence of new and changing store formats. Efforts should be made to determine accessibility, in terms of both proximity to a single location and variety of multiple locations within a specified buffer, from origins other than the home, and consider multipurpose trips and trip chaining. The measurement of food access will be critical for community-based approaches to meet dietary needs. Researchers must be willing to take the steps necessary for rigorous measurement of a dynamic food environment. PMID- 19285207 TI - Measurement of park and recreation environments that support physical activity in low-income communities of color: highlights of challenges and recommendations. AB - The capacity of public parks and recreation environments to promote physical activity for low-income communities of color is receiving increased attention from researchers and policymakers. As a result, several systems to measure park and recreation environments have been recently developed. Developing measures is important because they are critical to establishing key correlates and determinants that drive physical activity and inform intervention strategies. This paper briefly reviews recently developed approaches for measuring physical environments within public parks and recreation areas. It critiques the capacity of these approaches to advance an understanding of how parks and recreation settings contribute to physical activity in low-income communities of color. Residents of low-income communities of color are usually found to have lower physical activity, and this may be due partly to a disparity in access to parks and other recreation environments. Three primary recommendations are presented. First, future measurement tools should explicitly reflect inequality in the built environment in terms of availability and quality of parks and recreation areas. Second, measurement strategies should incorporate research on recreation activity and setting preferences important in low-income communities of color. Finally, the perceptions of residents of low-income communities of color should be reflected in measurement approaches. One strategy for incorporating the perceptions is community-based participatory research. The rapid development of high-quality tools for measuring parks and recreation environments is encouraging. However, existing measures should be tested and refined in varying social-ecologic conditions, and new tools should be developed specifically for nuances associated with low-income minority communities. PMID- 19285208 TI - Physical, consumer, and social aspects of measuring the food environment among diverse low-income populations. AB - Obesity and other diet-related chronic diseases are directly related to the food environment. We describe how to better assess the food environment in specific ethnic minority settings for designing and implementing interventions, based on a review of our previous work on the food environment in American Indian reservations, Canadian First Nations reserves, the Republic of the Marshall Islands, and inner-city Baltimore. The types of food stores available within each setting and the range of healthy foods available varied greatly across these geographic regions. In all settings, proximity to food stores/supermarkets, cost, and limited availability of healthful foods were common features, which limited access to health-promoting food options. Features specific to each population should be considered in an assessment of the food environment, including physical (e.g., openness of stores, mix of types of food sources); consumer (e.g., adequacy of the food supply, seasonal factors); and social (e.g., inter-household food sharing, perceptions of food quality, language differences) aspects. The food environments common in low-income ethnic subpopulations require special focus and consideration due to the vulnerability of the populations and to specific and unique aspects of each setting. PMID- 19285209 TI - Work group I: Measures of the food and physical activity environment: instruments. AB - A work group was convened to identify the core challenges, content gaps, and corresponding possible solutions for improving food- and physical activity environment instrumentation. Identified challenges included instrument proliferation, the scaling or grain of instruments and appropriate aggregation to the neighborhood or community level, and unknown sensitivity to change of most instruments. Solutions for addressing these challenges included establishing an interactive and real-time instrument repository, developing and enforcing high standards for instrument reporting, increasing community-researcher collaborations, and implementing surveillance of food and physical activity environment. Solid instrumentation will accelerate a better understanding of food and physical activity-environment effects on eating and physical activity behaviors. PMID- 19285210 TI - Work group II: Using Geographic Information Systems for enhancing research relevant to policy on diet, physical activity, and weight. AB - Geographic Information Systems (GIS) was a theme for one of the four workgroups convened for the Measures of the Food and Built Environment meeting held in Bethesda, Maryland in November 2007. This summary of group discussions frames several critical conceptual, methodologic, and data challenges regarding the use of GIS to enhance research relevant to policy on diet, physical activity, and weight. Broad recommendations are offered in five areas: (1) theoretical and conceptual development in framing place effects on health; (2) contextualizing people and spatial behavior in built environments and improving empirical representations of place; (3) geospatial data availability, quality, and standards; (4) privacy and confidentiality; and, (5) building capacity in GIS personnel and infrastructure. These topics are inter-related. Although our discussion focuses on issues relevant to the role of the built environment in diet and physical activity outcomes, our recommendations also are salient to health and environment research generally. PMID- 19285211 TI - Work group III: Methodologic issues in research on the food and physical activity environments: addressing data complexity. AB - Progress in transdisciplinary research addressing the health effects of the food and physical activity environments appears hampered by several methodologic obstacles, including: (1) the absence of clear, testable conceptual models; (2) slow adoption of practicable, rigorous research designs; (3) improper use of analytic techniques; and (4) concerns about ubiquitous measurement error. The consequence of such obstacles is that data collected as part of the typical study are more complex than need be. We offer diagnoses and recommendations from an NIH sponsored meeting that addressed core issues in food- and physical activity environment research. Recommendations include improved conceptual models and more elaborate theories, experimental thinking and increased attention to causal effect estimation, adoption of cross-validation techniques, use of existing measurement-error models, and increased support for methodologic research. PMID- 19285212 TI - Work group IV: Future directions for measures of the food and physical activity environments. AB - Much progress has been made in the past 5 to 10 years in measuring and understanding the impact of the food and physical activity environments on behavioral outcomes. Nevertheless, this research is in its infancy. A work group was convened to identify current evidence gaps and barriers in food and physical activity environments and policy research measures, and develop recommendations to guide future directions for measurement and methodologic research efforts. A nominal group process was used to determine six priority areas for food and physical activity environments and policy measures to move the field forward by 2015, including: (1) identify relevant factors in the food and physical activity environments to measure, including those most amenable to change; (2) improve understanding of mechanisms for relationships between the environment and physical activity, diet, and obesity; (3) develop simplified measures that are sensitive to change, valid for different population groups and settings, and responsive to changing trends; (4) evaluate natural experiments to improve understanding of food and physical activity environments and their impact on behaviors and weight; (5) establish surveillance systems to predict and track change over time; and (6) develop standards for adopting effective health promoting changes to the food and physical activity environments. The recommendations emanating from the work group highlight actions required to advance policy-relevant research related to food and physical activity environments. PMID- 19285213 TI - Measuring the food and physical activity environments: shaping the research agenda. PMID- 19285214 TI - Measuring physical activity environments: a brief history. AB - Physical activity is usually done in specific types of places, referred to as physical activity environments. These often include parks, trails, fitness centers, schools, and streets. In recent years, scientific interest has increased notably in measuring physical activity environments. The present paper provides an historical overview of the contributions of the health, planning, and leisure studies fields to the development of contemporary measures. The emphasis is on attributes of the built environment that can be affected by policies to contribute to the promotion of physical activity. Researchers from health fields assessed a wide variety of built environment variables expected to be related to recreational physical activity. Settings of interest were schools, workplaces, and recreation facilities, and most early measures used direct observation methods with demonstrated inter-observer reliability. Investigators from the city planning field evaluated aspects of community design expected to be related to people's ability to walk from homes to destinations. GIS was used to assess walkability defined by the 3Ds of residential density, land-use diversity, and pedestrian-oriented designs. Evaluating measures for reliability or validity was rarely done in the planning-related fields. Researchers in the leisure studies and recreation fields studied mainly people's use of leisure time rather than physical characteristics of parks and other recreation facilities. Although few measures of physical activity environments were developed, measures of aesthetic qualities are available. Each of these fields made unique contributions to the contemporary methods used to assess physical activity environments. PMID- 19285215 TI - Measuring food environments: a historical perspective. AB - Food and nutrition environments are believed to contribute to obesity and chronic diseases. There is a need for valid, reliable measures of nutrition environments. Familiarity with previous efforts to measure food and nutrition environments can help researchers and practitioners build on past accomplishments. This article describes sources of food-environment data, discusses how they have been used, and places the definition and measurement of food and nutrition environments in historical context. Review articles, agency websites, and peer-reviewed articles were the main sources of information. The review is organized around three main types of data sources identified as historic traditions: government, industry, and research. Types of data include archives, business monitoring records, surveys, observational assessments, and self-report surveys. Future development of clear, adaptable measures of food and nutrition environments will build on lessons of the past and will update and improve on past tools. PMID- 19285217 TI - The future of urologic oncology and surgeon-scientists: Proceedings of the World Urologic Oncology Federation, September 2007. PMID- 19285218 TI - The future of urologic oncology: "Proceduralists" or continued leaders? PMID- 19285216 TI - Measuring the built environment for physical activity: state of the science. AB - Physical inactivity is one of the most important public health issues in the U.S. and internationally. Increasingly, links are being identified between various elements of the physical-or built-environment and physical activity. To understand the impact of the built environment on physical activity, the development of high-quality measures is essential. Three categories of built environment data are being used: (1) perceived measures obtained by telephone interview or self-administered questionnaires; (2) observational measures obtained using systematic observational methods (audits); and (3) archival data sets that are often layered and analyzed with GIS. This review provides a critical assessment of these three types of built-environment measures relevant to the study of physical activity. Among perceived measures, 19 questionnaires were reviewed, ranging in length from 7 to 68 questions. Twenty audit tools were reviewed that cover community environments (i.e., neighborhoods, cities), parks, and trails. For GIS-derived measures, more than 50 studies were reviewed. A large degree of variability was found in the operationalization of common GIS measures, which include population density, land-use mix, access to recreational facilities, and street pattern. This first comprehensive examination of built environment measures demonstrates considerable progress over the past decade, showing diverse environmental variables available that use multiple modes of assessment. Most can be considered first-generation measures, so further development is needed. In particular, further research is needed to improve the technical quality of measures, understand the relevance to various population groups, and understand the utility of measures for science and public health. PMID- 19285219 TI - Urologic oncology in Asia. PMID- 19285220 TI - Oncologic urology in Latin America. PMID- 19285221 TI - Urologic oncology in southern Europe: Perceived problems and current plans. PMID- 19285222 TI - Urologic oncology in Germany. PMID- 19285223 TI - A medical oncologist's perspective on the treatment of patients with genitourinary malignancies. PMID- 19285224 TI - EAU-a work in progress-what comes next? PMID- 19285225 TI - Promoting surgeon-scientists in urologic oncology: The role of our journal. PMID- 19285226 TI - The urologist role in basic and clinical translational research: Personal reflections of a urologic scientist. PMID- 19285227 TI - Current environment surrounding clinical studies in Japan. PMID- 19285228 TI - Training of a surgeon-scientist. PMID- 19285229 TI - Renal cancer: Genetic directed therapy. PMID- 19285230 TI - The clinical implications of the genetics of renal cell carcinoma. AB - Over the last several decades, the advances in molecular genetics have elucidated kidney cancer gene pathways. Kidney cancer is a heterogeneous disorder. Each specific type of kidney cancer has its own histologic features, gene, and clinical course. Insight into the genetic basis of kidney cancer has been learned largely from the study of the familial or hereditary forms of kidney cancer. Extirpative surgery is currently the treatment of choice for kidney cancer that is confined to the kidney. Treatment for advanced or metastatic kidney cancer is a formidable challenge with the traditional therapies currently available. However, investigation of the Mendelian single-gene syndromes, like von Hippel Lindau (VHL: VHL gene), hereditary papillary renal carcinoma (HPRC: c-Met gene), Birt-Hogg-Dube (BHD: BHD gene), and hereditary leiomyomatosis renal cell cancer (HLRCC: fumarate hydratase gene) provides an opportunity to develop pathway specific therapies. Advances in molecular therapeutics offer novel treatment options for patients with advanced disease. PMID- 19285232 TI - Training the urologic oncologist of the future: where are the challenges? PMID- 19285233 TI - Training the urologic oncologist of the future: where are the challenges? AB - OBJECTIVE: Training the new generation of urologic oncologist from a surgical perspective poses unique challenges. The advent of minimally invasive surgical procedures coupled with the need to perform open surgical procedures has significantly increased the demands upon both the trainer and trainee. The learning and practice of complex procedures demand continuous improvements in surgical training programs. This review discusses some theoretical and practical issues to be considered for the successful and safe transmission of surgical skills in an era of increasing regulations. FINDINGS: Few systematic studies address this topic, leaving ample margin for research and improvement in this endeavor. It is the authors' opinion that mentorship remains the most significant and important component to successful surgical training today. The advent of simulation, virtual reality, and modular teaching represent (novel and important) advances in the field of surgical education. While some residency programs have incorporated these changes into their surgical training curriculum, this has not become widespread and the available literature remains at best sporadic. CONCLUSIONS: Mentorship remains an integral if not the most critical component to surgical training today. Other novel approaches to surgical training have developed and should be incorporated into the traditional concepts of mentorship training. This may become even more important with the advent of minimally invasive approaches to surgery. The vast majority of the studies published concur that the traditional model of "see one, do one, teach one" is not an optimal approach for training surgical skills. Socioeconomic changes are forcing the surgical community to rethink how they train residents and absorb new technologies. Adapting to the new demands posed on surgical educators represents a great challenge for the upcoming years. PMID- 19285234 TI - Critical elements in fellowship training. AB - In this article, we present the critical elements of fellowship training for the urologic oncologist. As an example, the Memorial Sloan-Kettering Cancer Center experience is outlined in detail, including the clinical experience, research curriculum, laparoscopic and minimally invasive exposure, and didactic lecture series. PMID- 19285235 TI - The hybrid of basic science and clinical training for the urologic oncologist: Necessity or waste? AB - OBJECTIVE: To examine the necessity and adequacy of basic science training for urologic oncology training programs. METHODS: Evaluated whether urology physician scientists are adequately trained in the basic sciences. RESULTS: The current urologic oncology training system does not adequately train physician scientists. We propose a major reform to define, train, and maintain the urology physician scientists. CONCLUSIONS: Urology physician scientists have played a major role in advancement of urologic oncology. Major reform is necessary, if we wish to continue to successfully train urologic oncology physician scientists. PMID- 19285236 TI - Minimally invasive surgical training: challenges and solutions. AB - Treatment options for urological malignancies continue to increase and include endoscopic, laparoscopic, robotic, and image-guided percutaneous techniques. This ever expanding array of technically demanding management options coupled with a static training paradigm introduces challenges to training the urological oncologist of the future. Minimally invasive learning opportunities continue to evolve, and include an intensive experience during residency, postgraduate short courses or mini-apprenticeships, and full time fellowship programs. Incorporation of large animal surgery and surgical simulators may help shorten the necessary learning curve. Ultimately, programs must provide an intense hands-on experience to trainees in all minimally invasive surgical aspects for optimal training. PMID- 19285237 TI - Robotic surgical training of the urologic oncologist. AB - Robotic-assisted surgery has become an increasingly popular approach to the treatment of a variety of urologic malignancies. The use of minimally invasive techniques for treatment of genitourinary cancers has evolved from conventional laparoscopy to the use of robotic-assisted instrumentation. Many questions remain regarding the safest and most effective way to teach robotic surgery to trainees. Work hour restrictions, medical and legal concerns, and the unique operative set up of the robotic system have made it increasingly difficult to provide "hands on" operative training to residents and fellows. We review the current literature regarding robotic surgical training, highlight potentially effective training strategies, and discuss future improvements in robotic surgical training of the urologic oncologist. PMID- 19285238 TI - Understanding the somatic-type malignant differentiation in male germ-cell cancer: a work in progress. Response to the article by Spiess et al., Malignant transformation of testicular teratoma: a chemoresistant phenotype. Urol Oncol 2008;26:595-9. PMID- 19285256 TI - The endocannabinoid system in metabolic control: a preface. PMID- 19285257 TI - An introduction to the endocannabinoid system: from the early to the latest concepts. AB - A rather complex and pleiotropic endogenous signalling system was discovered in the late 1990s, starting from studies on the mechanism of action of Delta(9) tetrahydrocannabinol, the major psychoactive principle of the hemp plant Cannabis sativa. This system includes: (1) at least two G-protein-coupled receptors, known as the cannabinoid CB(1) and CB(2) receptors; (2) the endogenous agonists at these receptors, known as endocannabinoids, of which anandamide and 2 arachidonoylglycerol are the best known; and (3) proteins and enzymes for the regulation of endocannabinoid levels and action at receptors. The number of the members of this endocannabinoid signalling system seems to be ever increasing as new non-CB(1) non-CB(2) receptors for endocannabinoids, endocannabinoid-related molecules with little activity at CB(1) and CB(2) receptors, and new enzymes for endocannabinoid biosynthesis and degradation are being identified every year. The complexity of the endocannabinoid system and of its physiological and pathological function is outlined in this introductory chapter, for a better understanding of the subsequent chapters in this special issue. PMID- 19285258 TI - The brain endocannabinoid system in the regulation of energy balance. AB - The role played by the endocannabinoid system in the regulation of energy balance is currently generating a great amount of interest among several groups of investigators. This interest in large part comes from the urgent need to develop anti-obesity and anti-cachexia drugs around target systems (such as the endocannabinoid system), which appears to be genuinely involved in energy balance regulation. When activated, the endocannabinoid system favors energy deposition through increasing energy intake and reducing energy expenditure. This system is activated in obesity and following food deprivation, which further supports its authentic function in energy balance regulation. The cannabinoid receptor type 1 (CB1), one of the two identified cannabinoid receptors, is expressed in energy balance brain structures that are also able to readily produce or inactivate N arachidonoyl ethanolamine (anandamide) and 2-arachidonoylglycerol (2AG), the most abundantly formed and released endocannabinoids. The brain action of endocannabinoid system on energy balance seems crucial and needs to be delineated in the context of the homeostatic and hedonic controls of food intake and energy expenditure. These controls require the coordinated interaction of the hypothalamus, brainstem and limbic system and it appears imperative to unravel those interplays. It is also critical to investigate the metabolic endocannabinoid system while considering the panoply of functions that the endocannabinoid system fulfills in the brain and other tissues. This article aims at reviewing the potential mechanisms whereby the brain endocannabinoid system influences the regulation energy balance. PMID- 19285259 TI - Role of acylethanolamides in the gastrointestinal tract with special reference to food intake and energy balance. AB - Acylethanolamides (AEs) are a group of lipids occurring in both plants and animals. The best-studied AEs are the endocannabinoid anandamide (AEA), the anti inflammatory compound palmitoylethanolamide (PEA), and the potent anorexigenic molecule oleoylethanolamide (OEA). AEs are biosynthesized in the gastrointestinal tract, and their levels may change in response to noxious stimuli, food deprivation or diet-induced obesity. The biological actions of AEs within the gut are not limited to the modulation of food intake and energy balance. For example, AEs exert potential beneficial effects in the regulation of intestinal motility, secretion, inflammation and cellular proliferation. Molecular targets of AEs, which have been identified in the gastrointestinal tract, include cannabinoid CB(1) and CB(2) receptors (activated by AEA), transient receptor potential vanilloid type 1 (TRPV1, activated by AEA and OEA), the nuclear receptor peroxisome proliferators-activated receptor-alpha (PPAR-alpha, activated by OEA and, to a less extent, by PEA), and the orphan G-coupled receptors GPR119 (activated by OEA) and GPR55 (activated by PEA and, with lower potency, by AEA and OEA). Modulation of AE levels in the gut may provide new pharmacological strategies not only for the treatment of feeding disorders but also for the prevention or cure of widespread intestinal diseases such as inflammatory bowel disease and colon cancer. PMID- 19285260 TI - The role of the endocannabinoid system in lipogenesis and fatty acid metabolism. AB - Endocannabinoids (ECs) regulate energy balance by modulating hypothalamic circuits controlling food intake and energy expenditure. However, convincing evidence has accumulated indicating that the EC system is present also in peripheral tissues, in particular in adipose tissue. Fat cells produce and are targets of ECs. Glucose uptake and lipoprotein lipase (LPL) activity, lipogenesis and adipogenesis are stimulated by ECs through cannabinoid 1 (CB1) receptors. Moreover, CB1 activation leads to a decreased mitochondrial biogenesis and function through inhibition of endothelial nitric oxide synthase (eNOS). All these effects are blocked by the CB1 antagonist rimonabant, suggesting that the weight-reducing effect of CB1 blockade is due not only to the transient suppression of food intake and reduction of lipogenesis but also to an increased mitochondrial biogenesis and oxidative metabolism which counteracts the inhibitory effects of ECs, levels of which are increased in fat tissues of obese rodents and humans. This review focuses on the role of ECs in adipose tissue metabolism, adipokine production, and interactions between ECs and peroxisome proliferator-activated receptors (PPARs) during adipogenesis. PMID- 19285261 TI - The role of the endocannabinoid system in liver diseases. AB - Endogenous cannabinoids (ECs) are ubiquitous lipid signaling molecules provided by a number of central and peripheral effects, which are mediated mainly by the specific receptors CB1 and CB2. In the last decade a considerable number of studies has shown that ECs and their receptors play an important role in the pathophysiology of liver diseases. The EC system is strongly up-regulated during chronic liver diseases. Until now it has been implicated in the pathogenesis of fatty liver disease associated with obesity, alcohol abuse, and hepatitis C, in the progression of fibrosis to cirrhosis, and in the development of portal hypertension, hyperdynamic circulatory syndrome and its complications, and cirrhotic cardiomyopathy. Furthermore, the EC system can participate in the pathogenesis of acute liver injury by modulating the mechanisms responsible for cell injury and inflammatory response. Thus, targeting the CB1 and CB2 receptors represents a potential therapeutic goal for the treatment of liver diseases. PMID- 19285262 TI - The role of the endocannabinoid system in the regulation of energy expenditure. AB - Endocannabinoids, a lipid-derived signaling system, regulate appetite and motivation to eat via effects in the hypothalamus and nucleus accumbens. Not all the effects of endocannabinoids on fat mass can be explained by the regulation of food intake alone. Endocannabinoids and their receptors are located in areas of the central nervous system and multiple peripheral tissues involved in the regulation of intermediary metabolism and energy expenditure. In addition to regulating food intake by both central and peripherally mediated effects, endocannabinoids modify glucose and lipid metabolism so as to promote energy storage via lipogenesis and reduce energy expenditure. The endocannabinoid system appears to be overactive in obesity and may serve to maintain fat mass and underlies some of the metabolic consequences of obesity. Inhibition of the cannabinoid type-1 receptor ameliorates the effects of endocannabinoids on food intake and energy metabolism; lipogenesis is inhibited, lipolysis, fatty acid oxidation and glucose uptake increase. PMID- 19285263 TI - The role of the pancreatic endocannabinoid system in glucose metabolism. AB - The endogenous cannabinoid system participates in the regulation of energy homeostasis, and this fact led to the identification of a new group of therapeutic agents for complicated obesity and diabetes. Cannabinoid receptor antagonists are now realities in clinical practice. The use of such antagonists for reducing body weight gain, lowering cholesterol and improving glucose homeostasis is based on the ability of the endocannabinoids to coordinately regulate energy homeostasis by interacting with central and peripheral targets, including adipose tissue, muscle, liver and endocrine pancreas. In this review we will analyse the presence of this system in the main cell types of the islets of Langerhans, as well as the physiological relevance of the endocannabinoids and parent acylethanolamides in hormone secretion and glucose homeostasis. We will also analyse the impact that these findings may have in clinical practice and the potential outcome of new therapeutic strategies for modulating glucose homeostasis and insulin/glucagon secretion. PMID- 19285264 TI - Use of cannabinoid CB1 receptor antagonists for the treatment of metabolic disorders. AB - Abdominal obesity is associated with numerous metabolic abnormalities, including insulin resistance, impaired glucose tolerance/type-2 diabetes, and atherogenic dyslipidaemia with low high-density lipoprotein (HDL) cholesterol, high triglycerides, and increased small dense low-density lipoprotein (LDL) cholesterol. A proportion of these metabolic disorders may be attributed to increased endocannabinoid activity. The selective cannabinoid 1 (CB1) receptor antagonist rimonabant has been shown to reduce body weight, waist circumference, insulin resistance, triglycerides, dense LDL, C-reactive protein (CRP), and blood pressure, and to increase HDL and adiponectin concentrations in both non-diabetic and diabetic overweight/obese patients. Besides an improvement in glucose tolerance in non-diabetic subjects, a reduction of 0.5-0.7% in haemoglobin A1C (HbA(1c)) levels was consistently observed in various groups of patients with type-2 diabetes. Almost half the metabolic changes could not be explained by weight loss, supporting direct peripheral effects of rimonabant. Ongoing studies should demonstrate whether improved metabolic disorders with CB1 receptor antagonists (rimonabant, taranabant, etc.) would translate into fewer cardiovascular complications among high-risk individuals. PMID- 19285265 TI - Use of cannabinoid receptor agonists in cancer therapy as palliative and curative agents. AB - Cannabinoids (the active components of Cannabis sativa) and their derivatives have received renewed interest in recent years due to their diverse pharmacological activities. In particular, cannabinoids offer potential applications as anti-tumour drugs, based on the ability of some members of this class of compounds to limit cell proliferation and to induce tumour-selective cell death. Although synthetic cannabinoids may have pro-tumour effects in vivo due to their immunosuppressive properties, predominantly inhibitory effects on tumour growth and migration, angiogenesis, metastasis, and also inflammation have been described. Emerging evidence suggests that agonists of cannabinoid receptors expressed by tumour cells may offer a novel strategy to treat cancer. In this chapter we review the more recent results generating interest in the field of cannabinoids and cancer, and provide novel suggestions for the development, exploration and use of cannabinoid agonists for cancer therapy, not only as palliative but also as curative drugs. PMID- 19285266 TI - Central side-effects of therapies based on CB1 cannabinoid receptor agonists and antagonists: focus on anxiety and depression. AB - Both agonists (e.g. Delta(9)-tetrahydrocannabinol, nabilone) and antagonists (e.g. rimonabant, taranabant) of the cannabinoid type-1 (CB(1)) receptor have been explored as therapeutic agents in diverse fields of medicine such as pain management and obesity with associated metabolic dysregulation, respectively. CB(1) receptors are widely distributed in the central nervous system and are involved in the modulation of emotion, stress and habituation responses, behaviours that are thought to be dysregulated in human psychiatric disorders. Accordingly, CB(1) receptor activation may, in some cases, precipitate episodes of psychosis and panic, while its inhibition may lead to behaviours reminiscent of depression and anxiety-related disorders. The present review discusses these side-effects, which have to be taken into account in the therapeutic exploitation of the endocannabinoid system. PMID- 19285267 TI - Cancer and thrombosis. PMID- 19285268 TI - Thrombosis and cancer: Trousseau syndrome revisited. AB - In this introductory chapter a story has been reviewed concerning the evolution of the concept of "cancer and thrombosis", since its first description by Armand Trousseau in 1865. From scattered reports on experimental material (tumor extracts) or on animal models of tumor/metastasis growth, through the progress of cell biology and experimental pharmacology, during the last 20 years interest has moved to clinical questions, such as: how to prevent and treat thrombosis, a frequent complication of both solid and hematologic malignancies? Has an occult cancer to be suspected in the majority of cases of idiopathic deep vein thrombosis? Do we need to prevent pharmacologically the occurrence of chemotherapy-associated thrombosis? Do anticoagulants have an impact on the natural history of some tumors? Why antiangiogenetic agents may be associated to a thrombotic risk? Presently, a continuous cross-talk between clinical results and experimental data is required to provide answers to these questions, taking advantage of a multidisciplinary approach to this old but still partially mysterious issue. PMID- 19285270 TI - Lessons from VTE registries: the RIETE experience. AB - Patients with venous thromboembolism (VTE) and active cancer are often excluded from clinical trials of anticoagulant therapy because of short life expectancy, inability or unwillingness to attend for regular laboratory monitoring during vitamin K antagonist therapy, or contraindications to therapy. As such, treatment regimens based on the results from clinical trials might not be suitable for many VTE patients with cancer. 'Registro Informatizado de la Enfermedad TromboEmbolica' (RIETE) was initiated in March 2001 to prospectively record the current clinical management of patients with symptomatic, objectively confirmed, acute VTE in Spanish hospitals. This chapter will summarize the most important findings published to date regarding the association between VTE and cancer. PMID- 19285269 TI - Epidemiology of cancer-related venous thromboembolism. AB - Recent studies have better defined the epidemiology of venous thromboembolism (VTE) in cancer patients. The incidence is highest in patients who have metastatic disease at the time of presentation and who have fast growing, biologically aggressive cancers associated with a poor prognosis. The incidence is also high in patients with haematological cancers. Other specific risk factors that affect the incidence of VTE include undergoing invasive neurosurgery, the number of underlying chronic co-morbid conditions, and being of Asian/Pacific Islander decent (lower incidence). The incidence is highest in the first few months after diagnosis, which may reflect the biology of the cancer or medical interventions such as major surgery or start of chemotherapy. The development of VTE is clearly associated with decreased survival, and this effect is greater among patients initially diagnosed with local- or regional-stage cancer compared with patients with metastatic cancer, probably because VTE reflects the presence of a biologically aggressive cancer. Finally, a small percentage of patients with idiopathic VTE and no clinical or laboratory evidence of cancer may harbour an aggressive but 'occult' malignancy likely causally linked to the development of VTE. PMID- 19285271 TI - Risk stratification for cancer-associated venous thromboembolism. AB - The risk of venous thromboembolism (VTE) is elevated in cancer, and thrombosis is the second leading cause of death in patients with malignancy. Many risk factors for cancer-associated thrombosis have been identified. These include patient associated factors such as age, obesity and medical comorbidities; cancer associated factors such as site and stage of cancer; and treatment-associated factors, particularly chemotherapy and hospitalization. In addition, several candidate biomarkers for cancer-associated thrombosis have been identified recently. Despite the high rate and significant impact of VTE in cancer outpatients, prior attempts at thromboprophylaxis in this population have not consistently demonstrated a benefit. This chapter will focus on risk stratification approaches, including a recently developed predictive model which can be used to identify those patients at highest risk. This model-based approach may have a significant impact on cancer-related morbidity, mortality and cost of care by directing targeted thromboprophylaxis in the future. PMID- 19285272 TI - Procoagulant mechanisms in tumour cells. AB - Pathogenesis of the prothrombotic state of cancer patients is due, at least in part, to the ability of cancer cells to activate the coagulation system. Several complex and not fully recognized interactions between the malignant cell and the clotting system promote a shift in the haemostatic balance compared with a procoagulant state. Tumour cells possess the capacity to interact with the haemostatic system in multiple ways. The principal mechanisms include the expression of haemostatic proteins by tumour cells, the production of inflammatory cytokines by tumour and/or host cells, and the direct adhesion of tumour cells to normal cells, including platelets, endothelial cells and monocytes. This chapter will summarize the prothrombotic mechanisms of tumour cells and their role in both coagulation and tumour progression. In particular, this chapter will focus on the capacity of tumour cells to promote activation of the coagulation system, and on the mechanisms by which clotting proteins may promote tumour growth and metastasis. PMID- 19285273 TI - Microparticles, thrombosis and cancer. AB - Microvesicles comprised of exosomes and microparticles are shed from both normal and malignant cells upon cell activation or apoptosis. Microvesicles promote clot formation, mediate pro-inflammatory processes, facilitate cell-to-cell interactions, transfer proteins and mRNA to cells, and induce cell signalling. Microparticles bearing tissue factor play a central role in coagulation initiation and thrombus formation. This chapter will review earlier studies which focus on the role of procoagulant microvesicles in cancer thrombogenicity, and discuss the effects of microvesicles on vascular cell dysfunction and angiogenesis. In addition, this chapter will present new findings which characterize the haemostatic balance of microparticles, and suggest a method that may potentially serve to predict a state of hypercoagulability in cancer patients. This chapter highlights the interplay between microvesicles, coagulation factors and cancer. PMID- 19285274 TI - Tissue factor in tumour progression. AB - The linkage between activation of the coagulation system and cancer is well established, as is deregulation of tissue factor (TF) by cancer cells, their vascular stroma and cancer-associated inflammatory cells. TF is no longer perceived as an 'alternative' coagulation factor, but rather as a central trigger of the coagulation cascade and an important cell-associated signalling receptor activated by factor VIIa, and interacting with several other regulatory entities, most notably protease-activated receptors (PAR-1 and PAR-2). Preclinical studies revealed the role of oncogenic transformation and tumour micro-environment as TF regulators in cancer, along with the impact of this receptor on gene expression, tumour growth, metastasis, angiogenesis and, possibly, formation of the cancer stem cell niche. Increasing interest surrounds the shedding of TF-containing microvesicles from cancer cells, their entry into the circulation and their role in the intercellular transfer of TF activity, cancer coagulopathy and other processes. Recent data also suggest differential roles of cell autonomous versus global effects of TF in various settings. Questions are raised regarding the consequences of TF expression by tumour cells themselves and by their associated host stroma. Progress in these areas may soon begin to impact on clinical practice and, as such, raises several important questions. Can TF be exploited as a therapeutic target in cancer? Where and when may this be safe and beneficial? Is expression of TF in various disease settings useful as a biomarker of cancer progression or the associated hypercoagulability? What clinical questions related to TF are especially worthy of further exploration, at present and in the near future? Some of these developments and questions will be discussed in this chapter. PMID- 19285275 TI - Heparanase coagulation and cancer progression. AB - Heparanase is an endo-beta-D-glucuronidase capable of cleaving heparan sulphate (HS) side chains of heparan sulphate proteoglycans on cell surfaces and the extracellular matrix; activity that is strongly implicated in tumour metastasis and angiogenesis. It has been shown that heparanase overexpression in human leukaemia, glioma and breast carcinoma cells results in a marked increase in tissue factor (TF) levels. In addition, TF was induced by exogenous addition of recombinant heparanase to tumour cells and primary endothelial cells; induction that was mediated by p38 phosphorylation and correlated with enhanced procoagulant activity. TF induction was further confirmed in transgenic mice overexpressing heparanase, and correlated with heparanase expression levels in leukaemia patients. Heparanase was also found to be involved in the regulation of tissue factor pathway inhibitor (TFPI). It has been shown that heparanase overexpression or exogenous addition induces a two- to three-fold increase in TFPI expression. Similarly, heparanase stimulated accumulation of TFPI in the cell culture medium. However, extracellular accumulation exceeded the observed increase in TFPI at the protein level, and appeared to be independent of HS and heparanase enzymatic activity. Instead, a physical interaction between heparanase and TFPI was demonstrated, suggesting a mechanism by which secreted heparanase interacts with TFPI on the cell surface, leading to dissociation of TFPI from the cell membrane and increased coagulation activity, thus further supporting the local prothrombotic function of heparanase. As heparins are strong inhibitors of heparanase, in view of the effect of heparanase on the TF/TFPI pathway, the role of anticoagulant activity of heparin may potentially be expanded. Taking into account the prometastatic and pro-angiogenic functions of heparanase, its overexpression in human malignancies and abundance in platelets, its involvement in the coagulation machinery is an intriguing novel arena for further research. PMID- 19285276 TI - Treatment of venous thromboembolism in cancer patients. AB - Venous thromboembolism (VTE) is a major cause of morbidity and mortality in patients with cancer. Monotherapy with low-molecular-weight heparin is the recommended first-line approach in cancer patients with newly diagnosed VTE, and is usually continued for a minimum of 3-6 months. Other management issues that require further research include optimal duration of anticoagulant therapy, treatment of recurrent VTE, the role of vena cava filters, the effects of VTE and its treatment on quality of life, and the impact of anticoagulants on survival. Newer anticoagulants hold promise in providing more effective and convenient treatment of VTE for this high-risk population, but further studies are awaited. PMID- 19285277 TI - Thrombosis in childhood acute lymphoblastic leukaemia: epidemiology, aetiology, diagnosis, prevention and treatment. AB - Acute lymphoblastic leukaemia (ALL) is the most common malignancy associated with venous thromboembolism (VTE) in children. The prevalence of symptomatic VTE ranges from 0% to 36%, and the variation can be explained, at least in part, by differences in chemotherapeutic protocols. The mechanism for increased risk of VTE is associated with alterations in the haemostatic system by use of L asparaginase (ASP) alone or in combination with vincristine or prednisone, presence of central venous lines (CVLs) and/or inherited thrombophilia. The children at greatest risk are generally those receiving Escherichia coli ASP concomitant with prednisone. The majority of symptomatic VTEs occur in the central nervous system or in the upper venous system. In the majority of cases, asymptomatic VTEs are associated with CVLs. External CVLs are affected more often than internal CVLs. Evidence-based guidelines on prevention and treatment guidelines for ALL-related VTE are lacking, and carefully designed clinical trials are needed urgently. PMID- 19285278 TI - Thrombosis associated with angiogenesis inhibitors. AB - Recent advances in the understanding of the pathogenesis of cancer have led to the introduction of a variety of biological agents with novel mechanisms of action into clinical trials and even into clinical practice. In particular, tumour-associated neoangiogenesis has become a major target for this new class of antineoplastic agents. Five anti-angiogenic agents (thalidomide, lenalidomide, bevacizumab, sunitinib, sorafenib) have already obtained US Food and Drug Administration approval for clinical use, and many others have entered clinical trials. Many new biological agents with anti-angiogenic properties appear to be associated with an increased risk for thrombosis and, paradoxically, bleeding. Although the mechanisms underlying the increased thromboembolic risk remain ill defined, the main hypothesis is that perturbation of tumour-associated endothelial cells can switch the endothelium from a naturally anticoagulant surface to a prothrombotic surface, thus mediating the activation of systemic coagulation in cancer patients, who are already more susceptible to thromboembolism due to their underlying disease. The toxicity profile differs between the anti-angiogenic agents. Thalidomide, lenalidomide, semaxibin (SU5416) and prinomastat have produced more venous thromboembolic complications, whereas bevacizumab, sunitinib, sorafenib and ZD6126 have been associated with a higher risk of arterial thromboembolism and, in particular, myocardial ischaemia. The observation of these vascular toxicities suggests the need to establish, in randomized clinical trials, the usefulness of thrombosis prophylaxis when anti angiogenic agents are used in cancer patients, especially when associated with chemotherapy. In addition, careful reporting of haemostatic complications during treatment with new anti-angiogenic drugs is warranted. PMID- 19285279 TI - Disseminated intravascular coagulation in cancer patients. AB - Disseminated intravascular coagulation (DIC) is a syndrome that may complicate a variety of diseases, including malignant disease. DIC is characterized by widespread, intravascular activation of coagulation (leading to intravascular fibrin deposition) and simultaneous consumption of coagulation factors and platelets (potentially resulting in bleeding). Clinically, DIC in cancer has, in general, a less fulminant presentation than the types of DIC complicating sepsis and trauma. A more gradual, but also more chronic, systemic activation of coagulation can proceed subclinically. Eventually, this process may lead to exhaustion of platelets and coagulation factors, and bleeding (e.g. at the site of the tumour) may be the first clinical symptom indicating the presence of DIC. In some cases, the clinical presentation of DIC in cancer may be reminiscent of thrombotic microangiopathies, which is understandable in view of the role of the endothelium in both conditions. The therapeutic cornerstone of DIC is treatment of the underlying disorder, but supportive treatment specifically aimed at the haemostatic system may be required. PMID- 19285280 TI - Thrombotic complications after haematopoietic stem cell transplantation: early and late effects. AB - Haematopoietic stem cell transplantation is currently the only curative option for many haematological malignancies, but is characterized by a wide spectrum of complications, including haemostatic changes. Bleeding and thrombotic events occur in the early and late phases after transplantation. In the early phase, thrombotic events have a variable clinical picture and present either as venous thrombosis, mainly at the site of central venous lines, veno-occlusive disease (also known as sinusoid occlusion syndrome) or transplant-associated microangiopathy. The latter two occur in the context of an acute graft-vs-host reaction, which involves various organs including the endothelium. In the late phase, years or decades after transplantation, thrombotic events present either as common venous thromboses or as arterial occlusions because of the development of premature atherosclerosis combined with diabetes, hypertension and dyslipidaemia, all of which are accelerated under the influence of the post transplant treatment. This chapter will discuss the incidence, possible causative associations and treatment options of early and late thrombotic events after haematopoietic stem cell transplantation. PMID- 19285281 TI - Antithrombotic therapy and survival in cancer patients. AB - Venous thromboembolism is a common complication in patients with malignant disease. The development of symptomatic thromboembolism is associated with a poor prognosis for patients with cancer. Retrospective analysis of studies comparing unfractionated heparin with low-molecular-weight heparin (LMWH) for the initial treatment of deep vein thrombosis indicated improved survival for cancer patients receiving LMWH therapy. Prospective studies exposing cancer patients to LMWH therapy, in the absence of established thrombosis, suggest that survival may be prolonged in these patients after LMWH exposure. Ongoing studies, with careful attention to distribution of prognostic variables for cancer outcome, will help to answer the question regarding whether LMWH may indeed be used for this indication. PMID- 19285282 TI - The coagulopathy of acute promyelocytic leukaemia revisited. AB - Since the initial description of the disease, the life-threatening coagulopathy associated with acute promyelocytic leukaemia (APL) has been the defining clinical characteristic. Historically, this uncommon subtype of acute myeloid leukaemia has been associated with a high mortality rate during induction therapy, most frequently attributable to haemorrhage. Since the introduction of all-trans retinoic acid (ATRA) into the therapy of all patients with APL, disease free survival and overall survival have improved dramatically, such that the disease is now highly curable. However, induction mortality remains a major problem and haemorrhage still accounts for the majority of such early deaths. Pathogenesis of the coagulopathy is complex and includes disseminated intravascular coagulation (DIC), fibrinolysis and proteolysis. As a result, while the predominant clinical manifestation of the coagulopathy is haemorrhage, thromboembolic events may occur both at presentation and during therapy. A major recent finding is the high expression of annexin II in the leukaemic cells from patients with APL. Annexin II is a protein with high affinity for plasminogen and tissue-type plasminogen activator (tPA), and also acts as a cofactor for plasminogen activation by tPA. As a result, both plasminogen and tPA are increased on the cell surface of the leukaemic cell, increasing plasmin activity. Annexin II is expressed in high amounts in cerebral microvascular endothelial cells, perhaps accounting for the relatively high incidence of intracranial haemorrhage in APL compared with other sites. Microparticles are cell-derived membrane fragments originating from normal cells or released from malignant cells involved in activating coagulation. Recent studies have found that microparticles containing tissue factor, tPA, plasminogen activator inhibitor-1 and annexin II have been found in the plasma of APL patients, suggesting a role in pathogenesis of the coagulopathy. Treatment of the coagulopathy remains primarily supportive. Aggressive transfusions of platelets and cryoprecipitate appear to be important. There is no clear role for the routine use of heparin or antifibrinolytic therapy. The most important factor may be the early introduction of ATRA at the first suspicion of a diagnosis of APL, before it is confirmed genetically. PMID- 19285283 TI - Different ways of practicing pharmaceutical care. PMID- 19285285 TI - Safeguarding against substandard/counterfeit drugs: mitigating a macroeconomic pandemic. AB - BACKGROUND: Counterfeiting and the sale of substandard pharmaceutical products can no longer be ignored. At 10% of global trade, counterfeiting is affecting many countries, causing serious downstream expenses and resource shortages. OBJECTIVE: To describe the nature and impact of drug product counterfeiting and substandard product sale and to present strategies that may have value in ameliorating these phenomena. METHODS: A literature review was conducted, supplemented by interviews of key leaders/experts in the field and the search of relevant web sites. All of the data were combined, integrated, and coordinated to present the complete picture of this problem. RESULTS: In addition to known corruption in some of the least developed countries, the trail through developed countries was detected. This report identifies means to detect faulty products and describes efforts toward resisting and ending these corrupt practices. CONCLUSIONS: Counterfeit drugs, if not stopped, can be responsible for a macroeconomic pandemic where major portions of some populations may be too ill to work and where the health sector resources are completely overwhelmed, as with the case of HIV/AIDS. PMID- 19285286 TI - Continuing effects of Medicare Part D on rural independent pharmacies who are the sole retail provider in their community. AB - BACKGROUND: The Medicare Prescription Drug, Improvement, and Modernization Act of 2003 established funding to allow Medicare beneficiaries to enroll in plans providing outpatient prescription drug coverage beginning in January 2006. The Medicare Part D program has changed the means by which beneficiaries purchase prescription drugs, impacting the business operations of pharmacies. OBJECTIVES: To describe the experiences of rural independently owned pharmacies that are the sole retail pharmacy in their community 1 year after implementation of Medicare Part D, in order to learn if the initial financial and administrative problems associated with the implementation of the program in 2006 resolved over time. METHODS: A semistructured interview protocol was used in telephone interviews with 51 pharmacist owners of rural sole community pharmacies in 27 states who were identified through a random sampling process. RESULTS: The sole community pharmacists interviewed continue to face challenges directly related to Medicare Part D. Dealing with Part D plans and working with patients during enrollment periods remains administratively burdensome. Reimbursement amounts, complexity of dealing with multiple plans, and timeliness of payments continue to be cited as problems which could threaten the viability of independently owned pharmacies who are the sole retail providers in their communities. CONCLUSIONS: Actions should be considered to help sole community pharmacies deal with the ongoing administrative and financial challenges of Part D. To ensure full choice for rural Medicare beneficiaries and full access to pharmaceuticals through the ongoing presence of a local pharmacy, the development of a mechanism to structure prescription reimbursement so that drug acquisition costs and related overhead are covered and a reasonable profit margin provided should be considered. Further study is needed to determine how existing policies and regulations can be modified to ensure reasonable access to pharmacy services for rural Medicare and Medicaid beneficiaries. PMID- 19285287 TI - Measuring patient satisfaction with diabetes disease state management services in community pharmacy. AB - BACKGROUND: Disease state management (DSM) programs for chronic conditions such as type 2 diabetes delivered by community pharmacists are a developing trend in health care service delivery. Although patient satisfaction with DSM services is an important indicator of service quality from the consumers' perspective, to date there is no valid and reliable instrument to enable its measurement in the context of a community pharmacy delivered service. OBJECTIVE: To develop and validate an instrument to measure patient satisfaction with diabetes disease state management (DDSM) services delivered by community pharmacists. METHODS: The DDSM questionnaire (DDSM-Q) was developed on the basis of 14 in-depth, semi structured interviews with consumers who had received diabetes care services in community pharmacy. Thirty-one questionnaire items were developed from the qualitative interview findings and also incorporated 16 modified items from an existing instrument, the Diabetes Measurement and Evaluation Tool. The DDSM-Q was sent to 160 participants of pharmacy diabetes care projects. Higher-order factor analysis was performed to extract factors of patient satisfaction. RESULTS: One hundred and fourteen questionnaires were returned, thus yielding a 71.3% response rate. Data from 108 out of 114 questionnaires were used to validate the DDSM-Q. Respondents had a mean age of 62 years and 61% were male. The 3 factors of patient satisfaction were interpreted as (1) satisfaction with the pharmacist's service ("Service"), (2) satisfaction with self-management ("Self-management"), and (3) satisfaction with knowledge and understanding of diabetes ("Knowledge"). The model fitted the data at moderate level (relative chi2 = 1.58, n = 108, adjusted goodness-of-fit index = 0.77, comparative fit index = 0.89, root mean square error of approximation = 0.07). The model was shown to have construct validity chi2(167) = 235.62, Bollen-Stine bootstrap P = .27). Reliabilities of the 3 factors were 0.92, 0.88, and 0.86, respectively. CONCLUSION: The results support the validity and reliability of the DDSM-Q as an instrument to measure patient satisfaction with DDSM services in community pharmacy. Further research will be needed to validate the instrument in different populations. PMID- 19285288 TI - The lay public's explicit and implicit definitions of drugs. AB - BACKGROUND: The research project: Public Beliefs about Medicine was initiated in 2001, aiming at exploring people's views, hopes, and fears with respect to drugs/medicines, now and in the future. OBJECTIVES: A part of the research project mapping public beliefs about medicines aimed at getting the public to discuss definitions of drugs/medicines to grasp their explicit and implicit definitions and understand their point of reference when using the term(s). METHODS: Eight focus groups were conducted in urban and rural Iceland, with 42 participants of both genders and varying age, a fifty-fifty mix of lay and educated groups. The interviews were transcribed verbatim and excerpts translated into English. RESULTS: The direct question about definition of drugs generally resulted in a consensus among the informants about drugs/medicines being used to combat diseases and/or symptoms. Some included only prescribed or physician recommended drugs in their definitions, others also over-the-counter products. Inclusion/exclusion of vitamins, food supplements, herbal remedies, functional foods, and illicit drugs was discussed without consensus and defining drugs in chemical terms or as interfering with bodily functions also occurred. New nuances emerged spontaneously later in the discussions, like viewing drugs as profit making products, preferably prudently marketed, wishing information about "new release" drugs for the informants' diseases, and being concerned about side effects, overuse, misuse, and abuse. Other implicit definitions included viewing drugs as a product-service package, or as a necessary evil and/or as products with increasing potency and associated future problems or considering drugs to exert unreliable minor influence. CONCLUSION: Professionals should be aware of the variety of lay people's drug definitions when discussing drugs/medicines with them to avoid misunderstandings caused by definition discrepancies. PMID- 19285289 TI - Cluster analysis of state Medicaid prescription drug benefit programs based on potential determinants of per capita drug expenditure. AB - BACKGROUND: Drug cost-containment strategies result in varied outcomes in different state Medicaid programs perhaps, because they differ in ways beyond the usual considerations of similarity among the states and their Medicaid populations. OBJECTIVE: To classify state Medicaid programs into homogenous groups based on potential determinants of their prescription drug expenditures. METHODS: Cluster analysis techniques were used to identify clusters of state Medicaid programs, and their characteristics, using variables identified as potential determinants of Medicaid prescription drug expenditures. RESULTS: Five distinct clusters were identified with memberships ranging from 1 to 24 states. Sociodemographic factors like race/ethnicity and elderly persons in Medicaid, severity of diseases, state population covered by Medicaid, population below poverty level, and high-school graduates; access factors like access to hospitals, pharmacies, physicians, and prenatal care; and policy factors like support for publicly funded health care were identified as key characteristics that distinguished among these clusters. Per capita prescription drug expenditure was also found to be significantly different among these clusters. CONCLUSION: Knowledge of clusters among state Medicaid programs and their characteristics could potentially increase the chances of success for Medicaid administrative policies and interventions at lower costs of targeted implementation. PMID- 19285290 TI - Perceptions, experiences, and expectations of physicians in hospital settings in Jordan regarding the role of the pharmacist. AB - BACKGROUND: To initiate a collaborative working relationship (CWR) between physicians and pharmacists, current physicians' perceptions, expectations, and experiences with pharmacists should be determined. OBJECTIVE: The purpose of this study was to investigate physicians' perceptions, expectations, and their actual experiences with pharmacists in hospital settings in Jordan. METHODS: A self administered questionnaire was delivered to 284 physicians selected randomly from 4 main hospitals in northern Jordan. The questionnaire was composed of 4 parts investigating physicians' expectations, experiences, and perceptions of the pharmacists. RESULTS: Two hundred and forty-five questionnaires were completed (response rate, 86.3%). More than half of the physicians were comfortable with pharmacists providing patient education. Similarly, just fewer than half (48.2%) were uncomfortable with pharmacists suggesting the use of prescription medications to patients. Most physicians (62.5%) expect the pharmacist to educate their patients about safe and appropriate use of drugs; however, approximately one third (33.9%) of them do not expect the pharmacist to be available for consultation during rounds. Approximately, 54% of the physicians agreed that pharmacists were always a reliable source of information. Nevertheless, only 28.2% agreed that pharmacists frequently inform them that their patients have experienced some problem with their medications. CONCLUSIONS: Physicians in hospitals in Jordan were more likely to accept or recognize traditional pharmacy services than newer clinical services. Increasing physician awareness of these clinical pharmacy skills will be an important step in developing CWRs. PMID- 19285291 TI - Characteristics of Illinois night shift work pharmacists. AB - BACKGROUND: Certain pharmacy sectors including 24-h community retailers and hospitals usually divide work schedules by shifts such as daytime, evening, and night shifts. The pharmacy literature dealing with the various types of alternative work schedules has not addressed the pharmacy night shift workforce specifically. OBJECTIVE: To obtain baseline information on night shift work pharmacists through analysis of the Illinois Pharmacist Compensation Survey 2005. METHODS: The biennial Pharmacist Compensation Survey, a 4-page self-administered questionnaire, was mailed in late 2005 to a random sample of pharmacists residing in Illinois, United States of America. Pharmacists who indicated that their gross base earnings included a shift differential were classified as shift work pharmacists. Descriptive statistics, Chi-square, and t tests were used to compare shift work pharmacists to non-shift work pharmacists. An alpha level of 0.05 was used to determine statistical significance. RESULTS: In Illinois, pharmacists receiving shift work differential pay were more likely to be women staff hospital pharmacists who work fewer weeks per year. Also, they spent significantly less time on business management activities and functions other than consultation, drug management, and dispensing. CONCLUSIONS: Night shift workers in pharmacy might be different from the nationwide shift workforce. Understanding the characteristics that best describe shift work pharmacists might help employers better tailor their recruitment efforts for this type of schedule. PMID- 19285292 TI - Assessment of self-medication practices in Assendabo town, Jimma zone, southwestern Ethiopia. AB - BACKGROUND: The actions taken for the treatment of illness or symptom of an illness vary depending on the perceptions and experiences of individuals and other factors. A significant portion of all care in illness is self-care. In many cases, self-medication is an important initial response to illness. Although some health-care providers attach negative connotations to it, the World Health Organization acknowledges the existence of a valid role of self-medication. OBJECTIVE: This study was aimed at assessing the magnitude, type, and factors of self-medication in Assendabo town, Jimma, southwestern Ethiopia. METHODS: A community-based cross-sectional study was conducted in Assendabo town during February and March 2006. Open-ended questionnaire was used to collect data by interviewing heads of households in the study population. The data collected were properly screened before they were analyzed. RESULTS: A total of 242 households with 1257 individuals were visited, of which 143 (11.4%) reported at least 1 episode of illness and of whom 56 (39%) used self-medication using both modern pharmaceuticals and traditional medicines. Low severity of illness was a major reason for practicing self-medication; 80.6% of self-medicating individuals had no information on potential drug adverse effect. About 55% of ill persons who treated themselves reported improvement in their condition. CONCLUSIONS: There is high prevalence of self-medication in Assendabo town. Lack of drug information and accessibility to over-the-counter drugs without any health professional guide contributed to the high incidence of self-medication. Enforcement of regulations in drug distribution and provision of appropriate health education to the community at large is critical. PMID- 19285293 TI - Multimethod research into policy changes in the pharmacy sector--the Nordic case. AB - This article aims to explain the nature of multimethod studies and to illustrate their role in pharmaceutical policy research. In the field of pharmaceutical policy research, methodological and theoretically sound evaluation is the main goal. Reflexive learning is required in order to address and resolve one of the important paradoxes of late modern societies, which is that while the increasing complexity of social systems progressively undermines notions of certainty in social knowledge, it simultaneously raises the stakes in relation to rational guidance of those systems. By reflecting over our own research processes, we identified the strengths and weaknesses of multimethod research. We present our research methods and the experiences of pharmaceutical policy changes from two separate evaluation studies, one from Iceland and the other from Denmark. In addition, examples from a third study in progress are included: a multimethod international comparison of recent changes in pharmaceutical policy in Iceland, Denmark and Norway. Based on our experiences and reflections, we identified four of the most important issues we encountered in carrying them out: The importance of doing research in context; Challenges of comparison and questions of compatibility of data--both qualitative and quantitative; The importance of doing these studies with a multidisciplinary team; Research in pharmaceutical policy is a fluid, ongoing process. PMID- 19285294 TI - Early parafoveal processing in reading Chinese sentences. AB - The possibility that during Chinese reading information is extracted at the beginning of the current fixation was examined in this study. Twenty-four participants read for comprehension while their eye movements were being recorded. A pretarget-target two-character word pair was embedded in each sentence and target word visibility was manipulated in two time intervals (initial 140 ms or after 140 ms) during pretarget viewing. Substantial beginning- and end-of-fixation preview effects were observed together with beginning-of fixation effects on the pretarget. Apparently parafoveal information at least at the character level can be extracted relatively early during ongoing fixations. Results are highly relevant for ongoing debates on spatially distributed linguistic processing and address fundamental questions about how the human mind solves the task of reading within the constraints of different writing systems. PMID- 19285295 TI - Haplotype-sharing analysis implicates chromosome 7q36 harboring DPP6 in familial idiopathic ventricular fibrillation. AB - Idiopathic Ventricular Fibrillation (IVF) is defined as spontaneous VF without any known structural or electrical heart disease. A family history is present in up to 20% of probands with the disorder, suggesting that at least a subset of IVF is hereditary. A genome-wide haplotype-sharing analysis was performed for identification of the responsible gene in three distantly related families in which multiple individuals died suddenly or were successfully resuscitated at young age. We identified a haplotype, on chromosome 7q36, that was conserved in these three families and was also shared by 7 of 42 independent IVF patients. The shared chromosomal segment harbors part of the DPP6 gene, which encodes a putative component of the transient outward current in the heart. We demonstrated a 20-fold increase in DPP6 mRNA levels in the myocardium of carriers as compared to controls. Clinical evaluation of 84 risk-haplotype carriers and 71 noncarriers revealed no ECG or structural parameters indicative of cardiac disease. Penetrance of IVF was high; 50% of risk-haplotype carriers experienced (aborted) sudden cardiac death before the age of 58 years. We propose DPP6 as a gene for IVF and increased DPP6 expression as the likely pathogenetic mechanism. PMID- 19285296 TI - Small stitches with small suture distances increase laparotomy closure strength. AB - BACKGROUND: There is no conclusive evidence which size of suture stitches and suture distance should be used to prevent burst abdomen and incisional hernia. METHODS: Thirty-eight porcine abdominal walls were removed immediately after death and divided into 2 groups: A and B (N = 19 each). Two suturing methods using double-loop polydioxanone were tested in 14-cm midline incisions: group A consisted of large stitches (1 cm) with a large suture distance (1 cm), and group B consisted of small stitches (.5 cm) with a small suture distance (.5 cm). RESULTS: The geometric mean tensile force in group B was significantly higher than in group A (787 N vs 534 N; P = .006). CONCLUSIONS: Small stitches with small suture distances achieve higher tensile forces than large stitches with large suture distances. Therefore, small stitches may be useful to prevent the development of a burst abdomen or an incisional hernia after midline incisions. PMID- 19285297 TI - Local wound exploration remains a valuable triage tool for the evaluation of anterior abdominal stab wounds. AB - BACKGROUND: Recent guidelines do not support local wound exploration (LWE) or diagnostic peritoneal lavage (DPL) in the evaluation of patients with anterior abdominal stab wounds (AASWs), favoring computed tomography scanning or serial examinations. In patients without immediate indications for laparotomy, we hypothesized that LWE/DPL would identify patients requiring surgery while limiting unnecessary hospital admissions. METHODS: Patients sustaining penetrating trauma at our level I trauma center over a 3-year period were reviewed. RESULTS: During the study period, 139 patients with AASW followed our LWE/DPL algorithm. Fifty-six patients had LWE without fascial penetration: 46 were discharged immediately, 10 required admission. Fifty-eight patients had fascial penetration on LWE but negative DPL: 37 were observed for less than 24 hours, 19 were observed for more than 24 hours, and 2 patients developed peritonitis requiring exploration. Twenty-five patients had positive LWE/DPL: 13 had therapeutic laparotomy, 12 had nontherapeutic laparotomy. CONCLUSIONS: Only 11% of patients with AASWs without overt indication for laparotomy require surgical care. LWE remains a valid method to exclude intra-abdominal injury and to eliminate hospitalization in more than one third of AASW patients. PMID- 19285298 TI - Adjuvant transcatheter arterial chemoembolization improves efficacy of hepatectomy for patients with hepatocellular carcinoma and portal vein tumor thrombus. AB - PURPOSE: To study the value of postoperative transcatheter arterial chemoembolization (TACE) to improve the efficacy of hepatectomy and tumor thrombus removal for patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT). EXPERIMENTAL DESIGN: From January 1996 to December 2004, 126 patients with HCC and PVTT were randomly assigned into 2 groups: a control group and a TACE group. The control group underwent liver resection combined with PVTT removal, and the TACE group underwent liver resection combined with adjuvant TACE after surgery. Survival time in the 2 groups was studied. RESULTS: The 2 groups were comparable with regard to all clinicopathologic data. The median survival time was 13 months (95% confidence interval [CI] 6.25 to 19.75 months) for the TACE group and 9 months (95% CI 6.90 to 11.10 months) for the control group. Estimated survival rates for 1, 3 and 5 years were better in the TACE group (50.9%, 33.8%, 21.5%; respectively) than the control group (33.3%, 17.0%, 8.5%, respectively; log rank P = .0094). CONCLUSIONS: Postoperative TACE enhances the effect of liver resection combined with PVTT removal for HCC patients with PVTT. PMID- 19285299 TI - Adverse events after radiofrequency ablation of unresectable liver tumors: a single-center experience. AB - BACKGROUND: Radiofrequency ablation (RFA) is a relatively new modality to treat liver tumors that is being incorporated into practice despite the fact that its risk profile has not been well described. METHODS: A retrospective cohort study, using structured chart review, on patients with liver tumor(s) was conducted from August 1998 to November 2006. Univariate and multivariate exploratory analyses were used to evaluate factors associated with adverse events. RESULTS: RFA procedures were performed on 196 patients (58% primary tumors, 24% colorectal metastases, and 18% other metastases). Twenty-three patients (12%) experienced serious adverse events. Multivariate analysis showed advanced age (>or=55 y), underlying liver disease, large tumor size (>4 cm), and concomitant procedure were associated with an increased risk of adverse events (P = .01, P < .01, P = .01, and P = .01, respectively). There were no in-hospital deaths. CONCLUSIONS: RFA was associated with acceptable morbidity and mortality. Factors associated with adverse events should be considered when counseling patients regarding RFA procedures. PMID- 19285300 TI - Feasibility of spinal anesthesia in laparoscopic ventral hernia repair: is it worth it? PMID- 19285301 TI - Outcomes of laparoscopic colorectal surgery in obese and nonobese patients: a case-matched study of 180 patients. AB - BACKGROUND: Because it has been suggested that obese patients may be at higher risk of morbidity and mortality after surgery, we conducted a prospective case matched study to compare outcomes of elective laparoscopic colorectal surgery in obese and nonobese patients. METHODS: Sixty-two consecutive nonselected obese patients (body mass index > or =30 kg/m(2)) were matched with 118 nonobese patients. Postsurgical mortality and morbidity were defined as in-hospital death and complications. RESULTS: Cardiopulmonary comorbidities were significantly more frequent in obese compared with nonobese patients (44% vs 24%, P < .01). Obesity was significantly associated with increased mean operating time (268 +/- 74 min vs 232 +/- 59 min, P < .001), and conversion rate (32% vs 14%, P < .01). The mortality rate was nil. The overall postsurgical morbidity rate (31% vs 19%, P = not significant) and mean hospital stay (11 +/- 10 days vs 9 +/- 8 days, P = not significant) were similar in obese and nonobese patients. CONCLUSIONS: The results of this large case-matched study suggest that laparoscopic approach for colorectal surgery is feasible and safe in obese patients. PMID- 19285303 TI - Risk factors for anastomotic leakage after resection for rectal cancer. PMID- 19285304 TI - Can intravenous lidocaine decrease postsurgical ileus and shorten hospital stay in elective bowel surgery? A pilot study and literature review. AB - BACKGROUND: This study examined whether systemic infusion of lidocaine, a local anesthetic with anti-inflammatory properties, can decrease surgical pain, length of postsurgical ileus, and hospital stay. METHODS: Twenty-two patients at a community hospital were randomized into 2 groups. Subjects were allocated to receive either lidocaine or a placebo infusion for the first 24 hours after surgery. RESULTS: Patients in the lidocaine group appeared to report less pain as reflected by a decrease in overall visual analogue scale pain scores 24 hours after surgery. The return of flatus after surgery was not considered significant (lidocaine 68.2 +/- 9.7 hours vs placebo 86.9 +/- 13.6 hours; P = .2802). The return of bowel movement after surgery was considered significant (lidocaine 88.3 +/- 6.08 hours vs placebo group 116 +/- 10.1 hours; P = .0286). The lidocaine group was discharged by mean day 3.76 +/- .24 versus placebo at mean day 4.93 +/- .42; P = .0277. CONCLUSIONS: Patients in the lidocaine group had bowel movements >24 hours earlier than those in the placebo group and were discharged earlier. PMID- 19285305 TI - Surgical time independently affected by surgical team size. AB - BACKGROUND: Team size and composition provide essential data for the study of operating room (OR) efficiency. METHODS: Laparoscopic procedures between July 2005 and July 2007 were reviewed retrospectively to record the number of OR personnel and the procedure time (PT). RESULTS: Of 399 procedures reviewed, 360 cases with complete data were analyzed. The average PT was 148 minutes. A mean of 8 different team members (range, 4-15) were involved. Surgeons and anesthesiologists stayed constant whereas the OR nurses were replaced more than once per procedure (mean, 4 nurses/procedure; range, 2-11). Multiple regression analysis revealed that both complexity of surgery and team size affected the PT significantly. When procedure complexity and patient condition were held constant, we found that adding 1 individual to a team predicted a 15.4-minutes increase in PT. CONCLUSIONS: The surgical team is a dynamic system with a large amount of member turnover. Efforts to improve OR efficiency should focus on decreasing surgical team size, limiting staff turnover, and enhancing communication between team members. PMID- 19285306 TI - Invited commentary on "impact of intraoperative behavior on surgical site infections" by Beldi et al. PMID- 19285307 TI - Impact of intraoperative behavior on surgical site infections. AB - BACKGROUND: The aim of this study was to identify intraoperative risk factors for surgical site infections (SSIs), which are accessible to interventions. We evaluated the effect of extensive intraoperative antiseptic measures and the impact of the behavior of members of the surgical team on SSIs. METHODS: Standard versus extensive antiseptic measures were randomly assigned in 1,032 surgical patients. The adherence to principles of asepsis by members of the surgical team was assessed prospectively. RESULTS: The rate of SSI was 14% with standard antiseptic measures and 15% with extensive measures (P = .581). Multivariate analysis identified following independent risk factors: lapses in discipline (odds ratio [OR] 2.02, confidence interval [CI] 1.05-3.88), intestinal anastomosis (OR 6.74, CI 3.42-13.30), duration of operation more than 3 hours (OR 3.34, CI 1.82-6.14), and body mass index >30 kg/m2 (OR 1.98, CI 1.22-3.20). CONCLUSION: Extensive measures of antisepsis did not reduce the incidence of SSI. A lapse to adhere to principles of asepsis was identified as an independent risk factor for the development of SSI (ClinicalTrials.gov number, NCT00555815). PMID- 19285308 TI - Synthesis of some O-, S- and N-glycosides of hept-2-ulopyranosonamides. AB - (O-Peracylated alpha-D-gluco- and -galacto-hept-2-ulopyranosylbromide)onamides gave the corresponding (alkyl beta-D-glyco-hept-2-ulopyranoside)onamides under Koenigs-Knorr conditions, and similar aryl glycosides were obtained with sodium phenolates; (aryl and hetaryl 2-thio-beta-D-gluco-hept-2-ulopyranoside)onamides were formed with thiophenols in the presence of K(2)CO(3) in acetone, and reactions with aniline in CH(2)Cl(2) furnished (N-phenyl beta-D-glyco-hept-2 ulopyranosylamine)onamides. Some deprotected derivatives of d-gluco configuration obtained by the Zemplen protocol showed no significant inhibition against rabbit muscle glycogen phosphorylase b. PMID- 19285309 TI - Toward the synthesis of fine chemicals from lactose: preparation of D-xylo and L lyxo-aldohexos-5-ulose derivatives. AB - The transformation of (5R)-2,6-di-O-benzyl-5-C-methoxy-beta-D-galactopyranosyl-(1 ->4)-2,3:5,6-di-O-isopropylidene-aldehydo-D-glucose dimethyl acetal (8) into partially protected derivatives of D-xylo- and L-lyxo-aldohexos-5-ulose has been reported, applying appropriate epimerisation methods to its 3'-O- and 4'-O protected alcoholic derivatives. PMID- 19285310 TI - Post-error slowing: an orienting account. AB - It is generally assumed that slowing after errors is a cognitive control effect reflecting more careful response strategies after errors. However, clinical data are not compatible with this explanation. We therefore consider two alternative explanations, one referring to the possibility of a persisting underlying problem and one on the basis of the low frequency of errors (orienting account). This latter hypothesis argues that infrequent events orient attention away from the task. Support for the orienting account was obtained in two experiments. Using a new experimental procedure, Experiment 1 demonstrated post-error slowing after infrequent errors and post-correct slowing after infrequent correct trials. In Experiment 2, slowing was observed following infrequent irrelevant tones replacing the feedback signals. PMID- 19285311 TI - A Bayesian formulation of behavioral control. AB - Helplessness, a belief that the world is not subject to behavioral control, has long been central to our understanding of depression, and has influenced cognitive theories, animal models and behavioral treatments. However, despite its importance, there is no fully accepted definition of helplessness or behavioral control in psychology or psychiatry, and the formal treatments in engineering appear to capture only limited aspects of the intuitive concepts. Here, we formalize controllability in terms of characteristics of prior distributions over affectively charged environments. We explore the relevance of this notion of control to reinforcement learning methods of optimising behavior in such environments and consider how apparently maladaptive beliefs can result from normative inference processes. These results are discussed with reference to depression and animal models thereof. PMID- 19285312 TI - The influence of framework design on the load-bearing capacity of laboratory-made inlay-retained fibre-reinforced composite fixed dental prostheses. AB - Delamination of the veneering composite is frequently encountered with fibre reinforced composite (FRC) fixed dental prosthesis (FDPs). The aim of this study is to evaluate the influence of framework design on the load-bearing capacity of laboratory-made three-unit inlay-retained FRC-FDPs. Inlay-retained FRC-FDPs replacing a lower first molar were constructed. Seven framework designs were evaluated: PFC, made of particulate filler composite (PFC) without fibre reinforcement; FRC1, one bundle of unidirectional FRC; FRC2, two bundles of unidirectional FRC; FRC3, two bundles of unidirectional FRC covered by two pieces of short unidirectional FRC placed perpendicular to the main framework; SFRC1, two bundles of unidirectional FRC covered by new experimental short random orientated FRC (S-FRC) and veneered with 1.5 mm of PFC; SFRC2, completely made of S-FRC; SFRC3, two bundles of unidirectional FRC covered by S-FRC. Load-bearing capacity was determined for two loading conditions (n = 6): central fossa loading and buccal cusp loading. FRC-FDPs with a modified framework design made of unidirectional FRC and S-FRC exhibited a significant higher load-bearing capacity (p<0.05) (927+/-74 N) than FRC-FDPs with a conventional framework design (609+/ 119 N) and PFC-FDPs (702+/-86 N). Central fossa loading allowed significant higher load-bearing capacities than buccal cusp loading. This study revealed that all S-FRC frameworks exhibited comparable or higher load-bearing capacity in comparison to an already established improved framework design. So S-FRC seems to be a viable material for improving the framework of FRC-FDPs. Highest load bearing capacity was observed with FRC frameworks made of a combination of unidirectional FRC and S-FRC. PMID- 19285313 TI - Theoretical analysis of alendronate and risedronate effects on canine vertebral remodeling and microdamage. AB - Bisphosphonates suppress bone remodeling activity, increase bone volume, and significantly reduce fracture risk in individuals with osteoporosis and other metabolic bone diseases. The objectives of the current study were to develop a mathematical model that simulates control and 1 year experimental results following bisphosphonate treatment (alendronate or risedronate) in the canine fourth lumbar vertebral body, validate the model by comparing simulation predictions to 3 year experimental results, and then use the model to predict potential long term effects of bisphosphonates on remodeling and microdamage accumulation. To investigate the effects of bisphosphonates on bone volume and microdamage, a mechanistic biological model was modified from previous versions to simulate remodeling in a representative volume of vertebral trabecular bone in dogs treated with various doses of alendronate or risedronate, including doses equivalent to those used for treatment of post-menopausal osteoporosis in humans. Bisphosphonates were assumed to affect remodeling by suppressing basic multicellular unit activation and reducing resorption area. Model simulation results for trabecular bone volume fraction, microdamage, and activation frequency following 1 year of bisphosphonate treatment are consistent with experimental measurements. The model predicts that trabecular bone volume initially increases rapidly with 1 year of bisphosphonate treatment, and continues to slowly rise between 1 and 3 years of treatment. The model also predicts that microdamage initially increases rapidly, 0.5-1.5-fold for alendronate or risedronate during the first year of treatment, and reaches its maximum value by 2.5 years before trending downward for all dosages. The model developed in this study suggests that increasing bone volume fraction with long term bisphosphonate treatment may sufficiently reduce strain and damage formation rate so that microdamage does not accumulate above that which is initiated in the first two years of treatment. PMID- 19285314 TI - Histopathological and immunohistochemical findings in the lungs of pigs infected experimentally with Mycoplasma hyopneumoniae. AB - Pigs were infected intranasally with Mycoplasma hyopneumoniae and killed at intervals ranging from 7 to 35 days post-infection (dpi). Histopathological changes consisted of (1) exudates in airways and alveolar lumina, (2) peribronchial and peribronchiolar lymphoid hyperplasia, and (3) enlargement of alveolar septa. These changes were particularly marked from 7 to 28dpi, coinciding with significant increases in the expression, detected immunohistochemically, of cytokines (IL-1alpha, IL-1beta, IL-8, TNF-alpha and INF gamma) and lymphoid markers (CD4+, CD8+, muramidase, IgG+, IgA+). Both the lesions and immunohistochemical signals declined in intensity beyond 35 days. PMID- 19285315 TI - Expression of genes encoding matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in normal and diseased canine mitral valves. AB - The pathogenesis of canine chronic valvular disease (CVD) is not fully characterized. The present study investigates the expression of genes encoding matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in normal and diseased mitral valves (MVs). Samples from normal (n=15) or diseased (n=10) canine MVs were subject to real-time polymerase chain reaction (PCR) for quantification of mRNA encoding MMP-1, -2, -9 and -14 and TIMP-2, -3 and -4. In normal valves there was low expression of mRNA encoding MMP-2, -9 and -14 and TIMP-3. In the valves from dogs with CVD there was significantly increased transcription of mRNA encoding MMP-1 and -14 and TIMP-2, -3 and -4, but no elevation in mRNA encoding MMP-2 and -9. MMPs and TIMPs are therefore likely to be involved in extracellular matrix metabolism in normal canine MVs and there are significant alterations in the expression of genes encoding these molecules during CVD. PMID- 19285316 TI - Thrombosis in Japanese patients with Fabry disease. AB - Fabry disease is an X-linked lysosomal storage disease resulting from deficient activity of the enzyme alpha-galactosidase (alpha-Gal) A. It has been postulated that the accumulation of globotriaosylceramide in the endothelial cells of blood vessels may lead to thrombosis of the brain and other tissues. Recently, enzyme replacement therapy (ERT) for Fabry disease is available. A high incidence of thrombotic accidents in Fabry disease has been postulated. However, a systemic study on thrombosis in cases of Fabry disease has not been undertaken. To clarify the incidence of thrombosis in Fabry disease, we screened 65 patients with Fabry disease (49 hemizygotes and 16 heterozygotes) from 39 unrelated Japanese families. We found that ten patients with Fabry disease (7 hemizygous males and 3 heterozygous females) had experienced thrombotic accidents, under 45-years-old in 8 cases. These 10 patients showed the gene mutations of classical Fabry disease. Nine of these thrombotic patients developed brain infarctions, one man who had the complication of recurrent thrombophlebitis, and the remaining woman showed central retinal artery occlusion and thrombophlebitis. We demonstrated a high incidence of thrombosis in Fabry disease (15%). ERT should be performed in patients not only in hemizygous males but also in heterozygous females and started at their early ages. PMID- 19285317 TI - [Fever, anemia and weight loss in the last 2 months in a woman of 21 years old]. PMID- 19285318 TI - [Oncologic risk counselling unit. Selection of risk groups and results]. AB - BACKGROUND AND OBJECTIVE: We evaluated the patients' selection and the results of the detection of mutations as well as the follow-up of families at a genetic counselling unit. PATIENTS AND METHOD: One hundred and fifty three patients were visited corresponding to 137 families: 77 of them were classified as high-risk group, 35 as moderate-risk and 25 as low-risk. The classification of patients in each group was made according to the recommendations of the guidelines and the "Oncoguia de cancer familiar" of the Departament de Salut de la Generalitat de Catalunya. RESULTS: With regard to familiar antecedents, patients of the high risk group had an average (standard deviation) of 2.80 (1.5) cases of breast and/or ovarian cancer, while it was 1.82 (0.75) in the moderate-risk group, and 1.05 (0.80) in the low-risk group. Thirty seven families of high-risk (51.9%) were studied for the detection of deleterious mutations in BRCA1/2. Of 37 completed studies, 5 were positive (one BRCA1 and one BRCA2), 3 had variants of unknown significance in BRCA2 and 25 (83.4%) had no deleterious mutation in BRCA1/2. CONCLUSIONS: A correct selection of patients was performed, because 16.6% of BRCA's studies was positive. This fact allows us to adapt a planning of screening. PMID- 19285319 TI - [Sustained remission following autologous recovery after non-myeloablative stem cell transplantation in a patient with Hodgkin's lymphoma]. PMID- 19285320 TI - [Methahemoglobinemia, methylene blue and antidote stockpile]. PMID- 19285321 TI - [Functional status as risk factor for mortality in patients with acute heart failure at the emergency department]. PMID- 19285322 TI - [Mortality from cardiovascular diseases in immigrants residing in Madrid]. AB - BACKGROUND AND OBJECTIVE: To estimate the mortality from cardiovascular diseases in immigrants residing in one of the regions of Spain with the highest immigration rate during the early years of the 21st century. POPULATION AND METHOD: The study included people aged 20 to 64 years of age residing in Madrid for the period 2000-2004. Mortality form cardiovascular disease in immigrants from different parts of the world was compared with the mortality in the native Spanish population. Mortality rates ratios adjusted for age, sex and per capita income in the area of residence were estimated. RESULTS: Immigrants from Sub Saharan Africa and from South America showed, respectively, the highest and the lowest mortality rate ratio of cardiovascular diseases and ischemic heart disease. Immigrants from the region of Central America and the Caribbean showed the highest mortality rate ratio of cerebrovascular disease. CONCLUSIONS: The pattern of cardiovascular disease mortality in immigrants residing in Madrid is quite similar to those found in studies made in other countries, and probably reflect the burden of disease in their places of origin. PMID- 19285323 TI - [Vitamin D as immunity element against infection]. PMID- 19285324 TI - [Incidence of type 1 diabetes mellitus in children: results from the population registry of the Madrid Region, 1997-2005]. AB - BACKGROUND AND OBJECTIVE: Incidence of type 1 diabetes mellitus (DM) varies importantly worldwide, including European countries, and even among regions within a country. The aim of this study is to describe the incidence of type 1 DM in the Madrid Region. MATERIAL AND METHOD: We included 1130 new cases of type 1 DM in children below 15 years of age, which were notified to the diabetes registry from January, 1997 to December, 2005. Case ascertainment was evaluated through the capture-recapture method. The data was analysed using Poisson regression models. RESULTS: The incidence rate in this period was 15.9/100,000 persons-years (95% confidence interval, 15.0-16.8). Completeness of ascertainment was 82%. The incidence by age group was 12.1 (0-4 years), 18.2 (5-9 years) and 17.4 by 100,000 persons-years (10-14 years). The incidence data showed no significant changes in the studied period. We observed a seasonal variation in the incidence, with the greatest incidence in the cold months. CONCLUSIONS: The estimated incidence of type 1 DM ranks in an intermediate position with regard to that estimated more recently in other Spanish regions. During the period 1997 2005, the incidence has maintained stable in the Madrid Region. PMID- 19285325 TI - [Motorocular syndromes due to neurogenic hyperactivity and their treatment]. AB - In this chapter we describe a variety of rare but clinically identifiable ocular motor syndromes, including ocular neuromyotonia, superior oblique myokymia, ocular motor synkinesis, third nerve palsy with cyclic spasms, and paroxysmal manifestations of multiple sclerosis. These syndromes share many characteristics. They result from neurogenic hyperactivity, causing episodic spasms of one or several extraocular muscles. The pathophysiology is not fully understood, but it usually includes both a focal and partial lesion of one of the ocular motor nerves and a central rearrangement of neuronal activity in the ocular motor nuclei. Treatment with membrane-stabilizing agents, such as carbamazepine, is usually effective to reduce the symptoms. The above-mentioned syndromes result from a number of different diseases. A proportion of apparently idiopathic cases may be related to a neurovascular compression syndrome. PMID- 19285326 TI - Molecular basis for cytokinin biosynthesis. AB - Cytokinins (CKs) are a group of phytohormones that play a crucial role in the regulation of plant growth and development. Identification of the enzymes and the corresponding genes that are involved in CK metabolism allowed us to understand how plants synthesize CKs and adjust CK activity to optimal levels. A major accomplishment toward these goals was the identification of genes for the first enzyme in the CK biosynthetic pathway, adenosine phosphate-isopentenyltransferase (IPT). In Arabidopsis thaliana and Agrobacterium tumefaciens, detailed analyses of IPTs were conducted through not only enzymatic characterization but also molecular structural approaches. These studies revealed the molecular basis for the Agrobacterium-origin of IPT used for the efficient biosynthesis of trans zeatin that promotes tumorigenesis in host plants. Another landmark in CK research was the identification of CYP735A as an enzyme that converts iP nucleotide to tZ-nucleotide. Furthermore, the identification of a CK-activating enzyme, LOG, which catalyzes a novel activation pathway, is a remarkable recent achievement in CK research. Collectively, these advances have revealed the complexity of the entire metabolic scheme for CK biosynthesis. PMID- 19285327 TI - [Glossary of terms and syndrome frequently used in epileptology]. PMID- 19285329 TI - Expression and characterization of human group C rotavirus virus-like particles in insect cells. AB - Group C rotavirus (GpC RV) is a causative agent of acute gastroenteritis in children and adults. We expressed the three major capsid proteins VP2, VP6 and VP7 of human GpC RV in baculovirus and demonstrated the self-assembly of VP2/6/7 or VP6/7 virus-like particles (VLPs) in insect cells. We examined a number of parameters, including the kinetics of protein synthesis in different cell lines and media, to optimize the most favorable conditions for the synthesis of recombinant viral proteins and the production of VLPs in Sf9 cells. Hyperimmune serum to VP2/6/7 and VP6/7 VLPs recognized individual recombinant proteins of human GpC RV by Western blot analysis. This serum also showed specific reactivities with the corresponding GpC VLPs but not GpA RV by using immune electron microscopy (IEM) and enzyme immunoassay (EIA). The ability to produce an unlimited amount of GpC RV antigen and the availability of high quality antibody will allow us to develop sensitive and specific diagnostic assays to better determine the epidemiology and disease burden of GpC RV in humans. PMID- 19285328 TI - Characterization of a myristoylated, monomeric HIV Gag protein. AB - The process of HIV assembly requires extensive homomultimerization of the Gag polyprotein on cellular membranes to generate the nascent particle bud. Here we generated a full-length, monomeric Gag polyprotein bearing mutations that eliminated multimerization in living cells as indicated by fluorescence resonance energy transfer (FRET). Monomeric Gag resembled non-myristoylated Gag in its weak membrane binding characteristics and lack of association with detergent-resistant membranes (DRMs or lipid rafts). Monomeric Gag failed to assemble virus-like particles, but was inefficiently rescued into particles by wildtype Gag through the influence of the matrix domain. The subcellular distribution of monomeric Gag was remarkably different than either non-myristoylated Gag or wildtype Gag. Monomeric Gag was found on intracellular membranes and at the plasma membrane, where it highlighted plasma membrane extensions and ruffles. This study indicates that monomeric Gag can traffic to assembly sites in the cell, where it interacts weakly with membranes. PMID- 19285330 TI - Identification and functional characterization of the left origin of lytic replication of murine gammaherpesvirus 68. AB - Murine gammaherpesvirus 68 (MHV-68) replicates robustly in cell culture, providing a model for studying viral genome replication during de novo infection of tumor-associated herpesviruses. We have previously identified a 1.25-kb origin of lytic replication (oriLyt) for MHV-68. To further investigate the molecular mechanism of viral genome replication, we first fine-mapped essential cis elements from this oriLyt fragment using a transposon-mediated high-density mutagenesis method. The result provided information for us to identify a second oriLyt located towards the left end of MHV-68 genome using a de novo infection replication assay. We further characterized this left oriLyt by scanning deletion analysis and site-directed mutations, and showed that several CCAAT motifs are essential for oriLyt function, whereas an AT-rich region enhances replication. However, GC-rich repeats are not important cis-element. Moreover, we identified a cellular transcription factor, NF-Y, which binds to CCAAT boxes in EMSA and associates with oriLyt in ChIP assay. Using a dominant negative expression plasmid, we demonstrated that NF-Y plays an important role in mediating MHV-68 genome replication during de novo infection. PMID- 19285331 TI - Extracellular polymeric substances (EPS) properties and their effects on membrane fouling in a submerged membrane bioreactor. AB - A pilot-scale submerged membrane bioreactor (MBR) for real municipal wastewater treatment was operated for over one year in order to investigate extracellular polymeric substances (EPS) properties and their role in membrane fouling. The components and properties of bound EPS were examined by the evaluation of mean oxidation state (MOS) of organic carbons, Fourier transform infrared (FT-IR) spectroscopy, three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy, and gel filtration chromatography (GFC), etc. Test results showed that MOS of organic carbons in the bound EPS was ranging from -0.14 to -0.51, and major components could be assessed as proteins and carbohydrates. FT-IR analysis confirmed the presence of proteins and carbohydrates in the bound EPS. The organic substances with fluorescence characteristics in the bound EPS were identified as proteins, visible humic acid-like substances and fulvic acid-like substances by EEM technology. GFC analysis demonstrated that EPS had part of higher MW molecules and a broader MW distribution than the influent wastewater. It was also found that a high shear stress imposed on mixed liquor could result in the release of EPS, which would in turn influence membrane fouling in MBRs. Bound EPS solution was observed to have a stronger potential of fouling than mixed liquor. During long-term operation of the MBR, bound EPS demonstrated positive correlations with membrane fouling while temperature was verified as a negative factor affecting EPS concentration. Compared to tightly bound EPS (TB EPS), loosely bound EPS (LB-EPS) showed more significant correlations with membrane fouling. This critical investigation would contribute towards a better understanding of the behavior, composition and fouling potential of EPS in MBR operation. PMID- 19285332 TI - Alkaline solubilization and microwave irradiation as a combined sludge disintegration and minimization method. AB - Commonly used pretreatment method of alkaline solubilization (using NaOH) and a relatively new technology of microwave (MW) irradiation (160 degrees C) were combined as a pretreatment method of waste activated sludge (WAS) in this study. First alkaline and MW pretreatment methods were examined separately, then their combination for different conditions was investigated in terms of their effect on COD solubilization, turbidity and capillary suction time (CST). For combined pretreatments, soluble COD to total COD ratio (SCOD/TCOD) of WAS increased from 0.005 (control) to 0.18, 0.27, 0.34 and 0.37 for combined methods of MW and pH 10, 11, 12 and 12.5, respectively. Deteriorated dewaterability due to alkaline pretreatment was also improved due to the incorporation of MW irradiation. Further, with small scale batch anaerobic reactors, pH-10, pH-12, MW (alone), MW+pH-10 and MW+pH-12 pretreated WAS samples were anaerobically digested. Highest total gas and methane productions were achieved with MW+pH-12 pretreatment with 16.3% and 18.9% improvements over control reactor, respectively. Finally the performance of MW+pH-12 pretreatment was examined with 2L anaerobic semi continuous reactors for 92 days and compared to that of the control reactors. These reactors were operated at an SRT of 15 days. After steady state, 43.5% and 55% improvements were obtained in respective daily total gas and methane productions. TS, VS and TCOD reductions were improved by 24.9%, 35.4% and 30.3%, respectively based on a relative calculation with respect to control reactors. This way combined alkaline-microwave treatment proved to be an effective sludge minimization method. Pretreated digested sludge had 22% improved dewaterability than unpretreated digested sludge. Higher SCOD and NH(3)-N concentrations were measured in the pretreated digested sludge supernatant; however, PO(4)-P concentration did not increase much. PMID- 19285333 TI - Using river distances in the space/time estimation of dissolved oxygen along two impaired river networks in New Jersey. AB - Understanding surface water quality is a critical step towards protecting human health and ecological stability. Because of resource deficiencies and the large number of river miles needing assessment, there is a need for a methodology that can accurately depict river water quality where data do not exist. The objective of this research is to implement a methodology that incorporates a river metric into the space/time analysis of dissolved oxygen data for two impaired river basins. An efficient algorithm is developed to calculate river distances within the BMElib statistical package for space/time geostatistics. We find that using a river distance in a space/time context leads to an appreciable 10% reduction in the overall estimation error, and results in maps of DO that are more realistic than those obtained using a Euclidean distance. As a result river distance is used in the subsequent non-attainment assessment of DO for two impaired river basins in New Jersey. PMID- 19285334 TI - Evaluation of optical brightener photodecay characteristics for detection of human fecal contamination. AB - Detection of optical brighteners by fluorometry combined with ultraviolet light (UV) exposure has been proposed as an inexpensive method for detection of human fecal contamination, but has received limited testing. This study evaluated the approach in southern California by applying it to a variety of detergents, sewage and septage samples from the region, as well as to natural stream water as a negative control. The concept of using UV exposure to differentiate fluorescence from natural organic matter proved valid, as the method produced no false positives. However, the method failed to detect half of the detergents tested in natural stream water at 5 microL/L, due to its conservative thresholds. This study identified a method modification that provides greater sensitivity by taking advantage of differences in the shape of photodecay curves between optical brighteners and natural organic matter. This method modification resulted in detection of all detergents, sewage at 1:10 dilution and septage at 1:100 dilution. Several caveats for its use remain, including our observation that the optical brightener signal degraded rapidly in strong sunlight. Additionally, there was low sensitivity for some environmentally friendly detergents, which does not present a problem on a community basis where a mix of detergents are used, but could be of concern for assessing septic inputs from individual homes. Still, the method is simple to employ in the field, yields rapid results and is useful as a low-cost initial screening tool. PMID- 19285335 TI - Survey on organochlorine pesticides, PCDD/Fs, dioxin-like PCBs and HCB in sediments from the Han river, Korea. AB - The contamination status of twelve persistent organic pollutants (POPs) on the Stockholm convention in the surface sediments of Han river, which is one of the largest river in the South Korea were investigated. Five organochlorine pesticides (OCPs) such as aldrin, dieldrin, endrin, mirex, and SigmaCHLs (alpha chlordane, gamma-chlordane, cis-nonachlor, trans-nonachlor, and heptachlor) were not detected in all sediment samples. The overall concentrations of DDTs, HCB, dioxin-like PCBs (DL-PCBs) and PCDD/Fs were in the range of 1.05-8.94microgkg(-1) (average value: 3.93microgkg(-1)), 0.485-3.73microgkg(-1) (1.48microgkg(-1)), 41.5-4,530ngkg(-1) (548ngkg(-1)), and 23.1-368ngkg(-1) (131ngkg(-1)), respectively. The principal source of HCB, DL-PCBs and PCDD/Fs was identified as a deposition in this study. For the DL-PCBs, it was also shown that commercial PCBs product (Kanechlor 500 and/or Aroclor 1254) plays a role as a source in sediment. PMID- 19285336 TI - Airborne chloronaphthalenes in Scots pine needles of Poland. AB - The amounts, profiles and origin of CNs (from triCNs to octaCN) sequestered in Scots pine needles collected from 25 spatially distant sites in Poland have been studied based on congener-specific data obtained after a several clean-up and fractionation steps and final HRGC/HRMS separation and determination. The absolute concentrations of CNs varied largely from site to site, i.e., by 15 fold. The sum of tri- to octaCN concentration at fifteen of the least contaminated sites ranged from 70 to 280 pg g(-1) ww, and at further eight sites were from 340 to 540 pg g(-1) ww, while at two the most contaminated were 1000 and 1100 pg g(-1)ww. There were some substantial similarities but also variations in triCN to octaCN homologue group profiles depending on the site. Among triCNs the isomers such as 1,2,4-/1,3,7-/1,4,6-triCNs (nos. 14/21/24) dominate in Scots pine needles. For majority of the sites examined 1,2,4-/1,3,7-/1,4,6-triCNs are also the major contributors to the bulk of CNs determined. Among tetraCNs isomer 1,2,5,8-tetraCN (no. 38) was dominant contributor at eighteen sites, while 1,2,4,6-/1,2,4,7-/1,2,5,7-tetraCN (nos. 33/34/37) at seven other sites. In the case of pentaCNs isomer 1,2,4,5,8-pentaCN (no. 59), was dominant contributor alone. Octachloronaphthalene frequently contributed substantially to the bulk of CNs. The Cluster Analysis and Principal Component Analysis did indicate that the compositional profiles of CNs found in Scots pine needles resemble somehow these found in the bottom ashes after coke and coal burning as well as of Halowax 1000 and 1099 formulations. PMID- 19285337 TI - Quantitative evaluation of perchlorethylene in groundwater before and after its oxidation by helical solid sorbent extraction and gas chromatography. AB - An integrated method of dynamic extraction of perchloroethylene (PCE) with polydimethylsiloxane (PDMS) helical solid sorbent followed by injection into a gas chromatograph was developed for the determination of the real concentration of PCE in groundwater before and after its degradation by oxidation with KMnO(4). The main parameters (agitation, temperature, salts, pH) that affect the extraction efficiency have been evaluated and optimized. The increase of the extracted amount of PCE due to the presence of the salts could be partially compensated by the opposite effect of the insoluble MnO(2), and of the presence of HCl and the global effect of the matrix would be less important for the reproducibility of the PCE extraction. The quantitative analysis of PCE was carried out with the external standard method using the calibration-curve technique. The limit of detection for PCE was found of 0.05 mg L(-1) using flame ionization detector. The solubility of PCE in water was studied by extraction of PCE with PDMS helical solid sorbent from standard solution with different amounts of PCE added into the same volume of water and was found to decrease with settling time, because the fine droplets created during the agitation of the mixture were slowly merged in drops on the bottle walls and were not transferred into the extraction vial for GC analysis. The limit of solubility was evaluated using a graphical method and was estimated at 224.2+/-15.7 mg L(-1) of PCE in water at 25 degrees C. PMID- 19285338 TI - A nationwide survey of NDMA in raw and drinking water in Japan. AB - A nationwide survey of N-nitrosodimethylamine (NDMA) in both raw and finished water samples from drinking water treatment plants (DWTPs) in Japan was conducted. NDMA was analyzed by solid-phase extraction (SPE) followed by ultra performance liquid chromatography (UPLC) coupled with tandem mass spectrometry (MS/MS). NDMA was detected in 15 of 31 raw water samples collected in the summer at concentrations up to 2.6 ng/L, and in 9 of 28 raw water samples collected in winter at concentrations up to 4.3 ng/L. The NDMA concentrations were higher in raw water samples collected from treatment plants with catchment areas that have high population densities. The NDMA concentrations were higher in river water samples collected from the east and west of Japan than in those collected from other areas. NDMA was detected in 10 of 31 finished samples collected in summer at reduced concentrations of up to 2.2 ng/L, while 5 of 28 finished samples collected in winter showed NDMA concentrations up to 10 ng/L. The highest NDMA levels were detected in finished water samples collected from the Yodo River basin DWTP, which uses ozonation. Furthermore, evaluation of the process water produced at six advanced water treatment plants was conducted. Influent from the Yodo River indicated that the NDMA concentration increased during ozonation to as high as 20 ng/L, and then decreased with subsequent biological activated carbon treatment. To our knowledge, this is the first nationwide evaluation of NDMA concentrations in water conducted in Japan to date. PMID- 19285339 TI - Glycoprobes as a tool for the study of lectins expressed on tumor cells. AB - Polyacrylamide glycoconjugates, Glyc-PAA, having various tags or labels are convenient tools for analysis of cellular lectins. Adaptation of such glycoprobes for flow cytometry allows us to reveal lectins expressed on cell surface and analyze their carbohydrate specificity as well as functionality. Localization of lectins is visualized by labeling of cells with fluorescein-tagged glycoprobes, Glyc-PAA-fluo, in combination with fluorescent microscopy techniques. Additionally, biotinylated glycoprobes can be immobilized on magnetic particles making it possible to separate a cell population according to its carbohydrate binding profile. Here, we exemplify application of glycoprobes in the study of cellular siglecs and galectins, as well as lectin patterning of tumor cells. The specificity of sialic acid binding membrane-anchored lectins, siglecs-1, -5, -7, 8 and -9 was determined using this methodology. To study the carbohydrate-binding profile of soluble galactoside-binding lectins, galectins-1 or -3, these were loaded on (initially galectin free) Raji cells and probed using Glyc-PAA-fluo. Lessons learned from this model system allowed us to study the galectin distribution pattern of tumors: cells obtained from mice carrying mammary adenocarcinoma or lymphoma were probed with Glyc-PAA-fluo using flow cytometry. Disaccharide 6OSuLacdiNAc was shown to be the most potent probe for adenocarcinoma cells, demonstrating that 6OSuLacdiNAc-binding molecules accumulate on cell surface in a patch-wise distribution. PMID- 19285340 TI - Effects of alcohol on the levels of parvalbumin in rat hearts. AB - Chronic excessive alcohol administration has been reported to be associated with diastolic dysfunction. Parvalbumin (PV) is a calcium-binding protein present in cardiac myocytes and involved in mediating relaxation. Therefore, alteration of PV levels may affect relaxation in cardiac myocytes. This study investigated the effects of alcohol administration on the levels of PV in the rat heart. Male Wistar rats weighing 200-250 g were divided into 2 groups: control (C) and alcohol-treated groups. The control group was provided with distilled water and the alcohol groups were provided with either a low dose (LD, 2g/kg) or high dose of ethanol (HD, 5 g/kg) once daily for 21 days, 3 months or 6 months. The PV levels in the ventricles were investigated by immunohistochemistry and Western blot analysis. In the 21-day ethanol-treated groups, parvalbumin immunoreactivity (PV-ir) and protein levels were not different when compared to the C, LD and HD groups. In the 3-month ethanol-treated groups, PV-ir and PV protein levels were decreased in both the LD and HD groups compared to that of the control group. In the 6-month ethanol-treated groups, PV-ir and PV protein levels decreased significantly in both the LD and HD groups (P<0.05). This indicates that short term ethanol treatment may not affect PV levels, whereas, long-term ethanol treatment clearly reduced PV levels. The decrease of PV was predominantly due to the direct toxic effects of alcohol rather than malabsorption caused by pathological changes in the duodenum and liver. The toxic effects of alcohol leading to a reduction of PV levels may lead to diastolic impairment. PMID- 19285341 TI - Differentiation of bone marrow-derived mesenchymal stem cells into multi-layered epidermis-like cells in 3D organotypic coculture. AB - The interactions of bone marrow-derived mesenchymal stem cells (MSCs) and their engrafted microenvironment are an integral part of signaling control of stem cell lineage commitment. We attempted to induce bone marrow-derived MSCs to undergo epidermal lineage differentiation by manipulating the biochemical, environmental and physical properties of culture conditions in an organotypic coculture model to simulate a skin-specific microenvironment. The induction medium was optimized by varying different biomolecular supplements in a basic stratification medium. A multi-layered epidermis-like structure was established when MSCs were cultured in an optimized induction medium on a contractible fibroblast-embedded collagen gel with an air-liquid interface. The commitment into epidermal lineage was further confirmed by the expression of early and intermediate epidermalization markers - keratin 10 and filaggrin in 90.67% and 80.51% of MSCs, respectively. This study not only highlights the possibility of in vitro control of MSCs into epidermal lineage, but also suggests the therapeutic potential of bone marrow-derived MSCs for skin regeneration. PMID- 19285342 TI - Metabolic scaling law for fetus and placenta. AB - The human fetal birth weight does not scale linearly with the weight of the placenta, but exhibits an allometric scaling consistent with Kleiber's Law for the basal metabolic rate. We discuss the possible causes of such scaling, and its clinical consequences. In particular, we show that the value of the scaling exponent is an indicator of a normal fetoplacental development. PMID- 19285343 TI - The oral toxicity of the transgenic Bt+CpTI cotton pollen to honeybees (Apis mellifera). AB - Transgenic insect-resistant cotton has been planted in China in a large scale and may have adverse impacts on honeybees. Pollens from the transgenic Cry1Ac+CpTI cotton Zhong-41 and the parental cotton Zhong-23 were collected from the field and their impacts on adult worker bees were assessed. Experimental results showed that Zhong-41 pollen had no acute oral toxic effect on worker bees. No significant differences were observed in the superoxide dismutase activity or in the longevity of worker bees fed with diets containing the two cotton pollens. The main reasons for the outcome may be the low expression level of the transgenic proteins Cry1Ac and CpTI in the pollen of Zhong-41 as well as the substantial equivalence in the amounts of gross protein and soluble saccharides for the two cotton pollens. The implications of these results are discussed and further work to be carried out is put forward. PMID- 19285344 TI - Effects of the insect growth regulator pyriproxyfen in a two-generation test with Chironomus riparius. AB - Pyriproxyfen a juvenile hormone analoga (JHA) was used in a two-generation test with Chironomus riparius (Diptera). The protocol is an extension of the OECD test methods to test duration from first-instar larvae in the parental (P) generation to emergence of midges in the filial (F1) generation. Test species were exposed to concentrations ranging from 0.001 to 0.1mg/L. The results showed significant adverse effects compared to the solvent control for emergence rate (p<0.001), development rate (p<0.001), fecundity (p<0.05) and fertility of egg ropes but not for the sex ratio. No observed effect concentrations (NOEC), respectively, lowest observed effect concentration (LOEC) values were lower for emergence rate and development rate in the F1 (0.003mg/L, respectively, 0.01mg/L each) than in the P generation (0.01mg/L, respectively, 0.03mg/L). The two-generation test provided additional information, which might be useful in the risk assessment of agrochemicals with potential endocrine effects. PMID- 19285345 TI - Subchronic exposure to nitrite, potassium and their combination on survival, growth, total haemocyte count and gill structure of juvenile blue swimmer crabs, Portunus pelagicus. AB - Among anthropogenic discharges, nitrite and potassium can simultaneously enter aquatic ecosystems at high loading rates which can threaten marine animals. The sensitivity of early juvenile blue swimmer crabs, Portunus pelagicus, to the subchronic exposure to nitrite, potassium and their combination was evaluated by measuring the survival, growth, total haemocyte count (THC) and gill histopathological changes. In all NaNO(2)-N and KNO(2)-N treatments the survival substantially decreased, many due to "molt death syndrome", along with significantly lower (p<0.01) specific growth rates (SGR). Although fewer deaths occurred in the KCl-K treatments, the SGR at the higher concentrations were significantly less (p<0.05) than the control. The gill histopathological changes following elevated NaNO(2)-N, KNO(2)-N and KCl-K exposure showed drastic but similar damage. In spite of a long term healing response, indicated by a significant hemolymph THC increase (p<0.01) and several gill lamellae modifications, early P. pelagicus juveniles are highly sensitive to elevated NO(2)-N levels. PMID- 19285346 TI - Abnormal response to failure in unmedicated major depression. AB - BACKGROUND: An aspect of neuropsychological impairment which has been linked specifically to depression is an abnormal response to failure. That is, a rapid deterioration of performance after receiving feedback that an error was made on the previous task. We aimed to examine this phenomenon in unmedicated, depressed outpatients. METHODS: Forty-four patients meeting DSM-IV criteria for major depression, all psychotropicmedication-free for at least six weeks, and 44 demographically matched, healthy control participants completed a computerised simultaneous/delayed matching-to-sample task (S/DMTS). RESULTS: Patients with depression were significantly less accurate than controls on the S/DMTS task. Both groups augmented their performance after an error had been made. The probability of making an error following an error was significantly greater in depressed compared with control participants, even when total number of errors was controlled for. Response latencies reduced significantly after an error had been made for both groups. LIMITATIONS: Both groups made relatively few errors. This reduced the power of analysis particularly when examining the effect of delay. CONCLUSIONS: The abnormal response to negative feedback previously identified in depressed samples was replicated in the current unmedicated, less severely depressed group. The impairment shown in the depressed sample may be due to a reduction in the motivating effect of an error compared with healthy controls. This has possible relevance to both neurobiological and psychological theories of depression. PMID- 19285347 TI - Depressive symptomatology is influenced by chronotypes. AB - BACKGROUND: Rhythm disturbances are a frequent clinical manifestation of depression. In recent years a possible relationship between depression and chronotypes has emerged. Specifically eveningness has been proposed as vulnerability factor. The aim of this study was to describe sleep features of depressed patients according to chronotypes and to explore possible associations with the clinical features of depressive episodes. METHODS: 100 patients diagnosed with Major Depressive Disorder according to the Mini International Neuropsychiatric Interview (MINI) were included (age: 34+/-11.74, range: 18-60 years; female/male:79/21). At admission the Hamilton Rating Scale for Depression (HRSD) was administered. Patients were also administered the Morningness Eveningness Questionnaire (MEQ), the Epworth Sleepiness Scale, the Athens Insomnia Scale and the Pittsburgh Sleep Quality Index. RESULTS: According to MEQ scores patients were classified in three groups: a) eveningness (n=18), b) neither (n=61) and c) morningness type (n=21). The age was different among chronotypes, being morningness-type patients older. The eveningness-type group showed higher scores in suicidal thoughts, more impaired work and activities, higher paranoid symptoms, higher scores on the anxiety cluster (HRSD), while the morningness-type group showed lower proportion of melancholic symptoms (MINI). We did not find association between sleep parameters and specific chronotypes. LIMITATIONS: The relatively small sample size and the concurrent assessment of chronotypes and depression may have biased our findings. CONCLUSIONS: Our data suggest the idea that chronotypes have an impact on depressive episodes features, with higher severity for the eveningness-type. PMID- 19285348 TI - Pediatric bipolar disorder in a Spanish sample: features before and at the time of diagnosis. AB - INTRODUCTION: Bipolar disorder (BD) often starts in childhood or adolescence. Diagnostic delay is common and may have a negative impact on treatment response and outcome. OBJECTIVES: To describe the clinical characteristics and symptoms of children with BD prior to their diagnosis and at the time of diagnosis in a sample in Spain. METHODS: We retrospectively reviewed the medical records of all children and adolescents (N=38) with a DSM-IV diagnosis of BD evaluated in the Child & Adolescent Psychiatry Unit, University of Navarra, over a 6-year period. We collected the DSM-IV symptoms of BD prior and at the time of diagnosis using the K-SADS-PL interview template. RESULTS: BD was diagnosed in close to 4% of clinic patients. Thirty (79%) were boys and 8 (21%) were girls; 17 (44.7%) had BD 1, 2 (5.3%) BD-2, and 19 (49.9%) BD-NOS. Median age at diagnosis was 13.9 (10.6;15.9). Delay of diagnosis was 1.5 (0.7;3.4) years. Symptoms of BD were similar to those reported in U.S. samples with high rates of severe irritability (94.6%) and psychiatric comorbidity: 92.1% of the BD children had at least one comorbid disorder and 18.4% had three comorbidities, most frequently ADHD (21%) and substance abuse (18.4%). CONCLUSIONS: Clinical findings in this Spanish sample of children with BD closely resembles those described in U.S. clinics. Diagnostic delay, as in the U.S., and frequent misdiagnosis may explain low prevalence estimates found outside the U.S. PMID- 19285349 TI - Perceived stigma among individuals with common mental disorders. AB - BACKGROUND: Severe mental disorders are associated with social distance from the general population, but there is lack of data on the stigma reported by individuals with common mental disorders. AIMS: To identify the correlates and the impact of stigma among individuals with common mental disorders. METHODS: Cross-sectional, household interview survey of 8796 representing the non institutionalized adults of Belgium, France, Germany, Italy, the Netherlands and Spain. Two perceived stigma questions (embarrassment and discrimination) were asked to respondents with significant disability. Health-related quality of life measured by the SF-12, work and activity limitation and social limitation were also assessed. RESULTS: Among the 815 participants with a 12-month mental disorder and significant disability, 14.8% had perceived stigma. Stigma was significantly associated with low education, being married/living with someone and being unemployed. Perceived stigma was associated with decreased quality of life (SF-12 PCS score -4.65; p<0.05), higher work and role limitation and higher social limitation. CONCLUSION: Individuals with mental disorders are more likely to report stigma if they have lower education, are married, or are unemployed. Perceived stigma is associated with considerably decrease in quality of life and role functioning. Health professionals and society at large must be aware of these findings, which suggest that fighting stigma should be a public health priority. PMID- 19285350 TI - Comparable performance of patients with obsessive-compulsive disorder (OCD) and healthy controls for verbal and nonverbal memory accuracy and confidence: time to forget the forgetfulness hypothesis of OCD? AB - The memory deficit or forgetfulness hypothesis of obsessive-compulsive disorder (OCD) has received considerable attention and empirical effort over the past decades. The present study aimed to provide a fair test of its various formulations: (1) memory dysfunction in OCD is ubiquitous, that is, manifests irrespective of modality and material; (2) memory dysfunction is found for nonverbal but not verbal material, (3) memory dysfunction is secondary to executive impairment; and (4) memory dysfunction affects meta-memory rather than memory accuracy. Participants comprised 43 OCD patients and 46 healthy controls who were tested on the Picture Word Memory Test (PWMT), which provides several unconfounded parameters for nonverbal and verbal memory accuracy and confidence measures across different time-points. In addition, the Trail-Making Test B was administered to test assumption number 3. Replicating earlier work of our group, samples displayed similar performance on all indices. None of the different formulations of the memory deficit hypothesis were supported. In view of waning evidence for a global memory deficit in OCD, neuropsychological research on OCD should more thoroughly investigate moderators and triggers of occasional instances of impaired performance, particularly cognitive biases such as perfectionism and an inflated sense of responsibility. PMID- 19285351 TI - Examining executive functioning in children with autism spectrum disorder, attention deficit hyperactivity disorder and typical development. AB - Executive functioning (EF) is an overarching term that refers to neuropsychological processes that enable physical, cognitive, and emotional self control. Deficits in EF are often present in neurodevelopmental disorders, but examinations of the specificity of EF deficits and direct comparisons across disorders are rare. The current study investigated EF in 7- to 12-year-old children with autism spectrum disorder (ASD), attention deficit hyperactivity disorder (ADHD) and typical development using a comprehensive battery of measures assessing EF, including response inhibition, working memory, cognitive flexibility, planning, fluency and vigilance. The ADHD group exhibited deficits in vigilance, inhibition and working memory relative to the typical group; however, they did not consistently demonstrate problems on the remaining EF measures. Children with ASD showed significant deficits in vigilance compared with the typical group, and significant differences in response inhibition, cognitive flexibility/switching, and working memory compared with both groups. These results lend support for previous findings that show children with autism demonstrate generalized and profound impairment in EF. In addition, the observed deficits in vigilance and inhibitory control suggest that a significant number of children with ASD present with cognitive profiles consistent with ADHD. PMID- 19285352 TI - Stent thrombosis: nightmare of the interventional cardiology era. AB - Coronary stents have marked an era in the interventional cardiology and have significantly decreased the rates of acute restenosis. Although diminished rates of restenosis have been observed with stents with respect to balloon angioplasty, restenosis is still a major problem of the interventional procedures. Stent thrombosis (ST) after percutaneous coronary intervention (PCI) is a rare and usually poor prognostic event that might result in myocardial infarction (MI) and sudden death. We wanted to summarize the main features of the issue. PMID- 19285353 TI - Prognostic value of CA125 in advanced heart failure patients. AB - INTRODUCTION: Serum levels of CA125 are often high in advanced heart failure (AHF) patients. AIM: To determine the predictive value of CA125 in forecasting the occurrence of death or cardiac transplantation in an AHF population. METHODS: 88 AHF patients referred for heart transplantation were divided into 2 groups based on CA125 levels: normal (CA125<38 U/mL) and elevated (> or = 38 U/mL). Events (death or heart transplant) were monitored over a period of 13+/-7 months after CA125 determination. RESULTS: Patients with elevated CA125 (n=65) had significantly lower blood pressure, body mass index, serum sodium and peak exercise oxygen consumption, while B-type natriuretic peptide levels were significantly higher. The combined primary endpoint (death or heart transplant) rate was 39.4% and 62.3% in normal and elevated CA125 groups, respectively (p=0.029). Multivariate regression analysis showed that CA125 and sodium levels were the only independent predictors of the combined endpoint. CONCLUSION: In AHF patients, plasma CA125 was an effective prognostic marker. Its determination may contribute to better risk stratification in this population. PMID- 19285354 TI - Can FDG-PET assist in radiotherapy target volume definition of metastatic lymph nodes in head-and-neck cancer? AB - BACKGROUND AND PURPOSE: The role of FDG-PET in radiotherapy target volume definition of the neck was evaluated by comparing eight methods of FDG-PET segmentation to the current CT-based practice of lymph node assessment in head and-neck cancer patients. MATERIALS AND METHODS: Seventy-eight head-and-neck cancer patients underwent coregistered CT- and FDG-PET scans. Lymph nodes were classified as "enlarged" if the shortest axial diameter on CT was 10mm, and as "marginally enlarged" if it was 7-10mm. Subsequently, lymph nodes were assessed on FDG-PET applying eight segmentation methods: visual interpretation (PET(VIS)), applying fixed thresholds at a standardized uptake value (SUV) of 2.5 and at 40% and 50% of the maximum signal intensity of the primary tumor (PET(SUV), PET(40%), PET(50%)) and applying a variable threshold based on the signal-to-background ratio (PET(SBR)). Finally, PET(40%N), PET(50%N) and PET(SBRN) were acquired using the signal of the lymph node as the threshold reference. RESULTS: Of 108 nodes classified as "enlarged" on CT, 75% were also identified by PET(VIS), 59% by PET(40%), 43% by PET(50%) and 43% by PET(SBR). Of 100 nodes classified as "marginally enlarged", only a minority were visualized by FDG-PET. The respective numbers were 26%, 10%, 7% and 8% for PET(VIS), PET(40%), PET(50%) and PET(SBR). PET(40%N), PET(50%N) and PET(SBRN), respectively, identified 66%, 82% and 96% of the PET(VIS)-positive nodes. CONCLUSIONS: Many lymph nodes that are enlarged and considered metastatic by standard CT-based criteria appear to be negative on FDG PET scan. Alternately, a small proportion of marginally enlarged nodes are positive on FDG-PET scan. However, the results are largely dependent on the PET segmentation tool used, and until proper validation FDG-PET is not recommended for target volume definition of metastatic lymph nodes in routine practice. PMID- 19285355 TI - Boron neutron capture therapy for newly diagnosed glioblastoma. AB - PURPOSE: The efficacy, safety, and dose distribution of neutron capture therapy (NCT) were evaluated in 15 patients with newly diagnosed glioblastoma. METHODS AND MATERIALS: Seven patients received intraoperative NCT (protocol-1) and eight patients received external beam NCT (protocol-2). Sulfhydryl borane (5 g/body) was administered intravenously. Additionally, p-dihydroxyboryl-phenylalanine (250 mg/kg) was given in protocol-2. The external beam NCT was combined with fractionated photon irradiation. RESULTS: Four of 15 patients were alive at analysis for a mean follow-up time from diagnosis of 23.0M. Twelve of the 15 patients were followed up for more than one year, and 10 (83.3%) of the 12 patients maintained their Karnofsky Performance Status (KPS; 90 in eight patients and 100 in two patients) at 12 months. The median overall survival and the time to tumor progression (TTP) for all patients were 25.7 and 11.9 M, respectively. There was no difference in TTP between the protocol-1 (12.0 M) and protocol-2 (11.9 M). The 1- and 2-year survival rates were 80.0% and 53.3%, respectively. Three protocol-1 patients and one protocol-2 patient suffered transient orbital swelling accompanied by double vision (Grade 2); one of the three protocol-1 patients suffered post-epileptic brain swelling (Grade 4) requiring surgical intervention. CONCLUSION: It is suggested that NCT is effective for survival of newly diagnosed glioblastoma with acceptable adverse effects. Because of the limitation of the present NCT pilot study without the contemporary control arm, it is unconvincing whether the neutron capture reaction led to distinct survival benefits, and further optimized studies on less invasive external beam NCT in large series of patients are warranted. PMID- 19285356 TI - Long-term outcome of high-precision radiotherapy in patients with brain stem gliomas: results from a difficult-to-treat patient population using fractionated stereotactic radiotherapy. AB - INTRODUCTION: To assess long-term outcome in 85 patients with brain stem gliomas treated with fractionated stereotactic radiation therapy (FSRT). PATIENT AND METHODS: Thirty-nine patients were females, and 46 were males. Median age at primary diagnosis was 26 years. Thirty-one patients were younger than 18 years. Histopathological examination confirmed a low-grade glioma in 57 patients. Of the group of high-grade gliomas, six were anaplastic astrocytomas, and two were classified as glioblastoma. Radiation therapy was performed as FSRT. The median target volume was 101 ml. We applied a median dose of 54 Gy in conventional fractionation of 1.8 Gy. In seven of 85 patients (8%) FSRT was performed as re irradiation. RESULTS: The median follow-up time was 42 months. Median overall survival (OS) was 81 months. OS rates were 77% at 12 months, 70% at 24 months, and 63% at 36 months. Significant impact on OS could be shown for pilocytic histology, age, neurosurgical resection as well as for the presence of cyst on MR imaging. Median progression-free survival (PFS) after FSRT was 52 months. PFS rates at 12 months were 70%, and 63% and 58% at 24 and 36 months, respectively. Histology, partial neurosurgical resection and the duration of symptoms could be identified as significant prognostic factors. CONCLUSION: Long-term outcome of FSRT in patients with brain stem gliomas is acceptable with low rates of side effects. Significant impact on outcome could be shown for histology, age, extent of neurosurgical resection as well as for cyst formation. No dose-response relationship could be observed. PMID- 19285357 TI - On the origin of heterogeneity in (preservation) resistance of Bacillus spores: input for a 'systems' analysis approach of bacterial spore outgrowth. AB - Bacterial spores are the ultimate (stress) 'survival capsules'. They allow strains from the Bacillus and Clostridium species to survive harsh environmental conditions. In addition to the decision to enter sporulation the decision to do the reverse (germinate) is also a decisive event after which there is no return. Generally it is observed that the behaviour of spores towards the environment is not homogeneous. In fact in many cases it is even quite heterogeneous, certainly upon subjecting the spores to a thermal stress treatment. Genome information coupled to high resolution single-cell analysis techniques allow us currently to analyse signalling events of individual cells. In the area of food preservation the next challenge is to couple the newly acquired mechanistic data to the physiologically observed heterogeneity in spore behaviour. The current paper will introduce the background of physiological heterogeneity while discussing the molecular processes that likely contribute to the observed heterogeneity in outgrowth. The discussion is set in the framework of contemporary and future needs for single-cell data integration in order to enhance the mechanistic basis of food preservation and spoilage models targeting bacterial spores. PMID- 19285358 TI - Enhanced resveratrol accumulation in rolB transgenic cultures of Vitis amurensis correlates with unusual changes in CDPK gene expression. AB - It has been established that transformation of Vitis amurensis callus culture with the plant oncogene rolB of Agrobacterium rhizogenes results in a high level of resveratrol production in the transformed culture. In the present report, we investigated two rolB transgenic V. amurensis cell cultures with different levels of rolB expression and resveratrol production. We examined whether the calcium ion flux and later steps of the calcium-mediated signal transduction pathway play a role in resveratrol biosynthesis in the rolB transgenic cultures. It has been shown that the calcium channel blockers, LaCl(3), verapamil, and niflumic acid, significantly reduced the accumulation of resveratrol in the rolB transgenic cultures. The number of the calcium-dependent protein kinase (CDPK) transcript variants and abundance of some of the transcripts were considerably altered in the rolB transgenic cell cultures, as revealed by frequency analysis of RT-PCR products and real-time PCR. Some unusual CDPK transcripts with deletions and insertions in the kinase domain were isolated from cDNA probes of rolB transformed cells. These results suggest that active resveratrol biosynthesis in rolB transgenic cultures of V. amurensis is Ca2+ dependent. We propose that the rolB gene has an important role in regulation of calcium-dependent transduction pathways in transformed cells. PMID- 19285359 TI - Evaluation of colonisation of peripheral venous catheters inserted by prehospital emergency service teams (SMUR) in France. AB - There are no data in the literature on colonisation of peripheral venous catheters (PVCs) inserted by French prehospital emergency and resuscitation service teams (SMUR). In a descriptive and prospective study we evaluated colonisation of PVCs inserted and managed solely by a SMUR team. A total of 171 PVCs were analysed. Bacteriological results were positive for seven catheters (4.09%/threshold=10(2)cfu/mL). Our analyses of the characteristics of patients and interventions show a significantly higher number of elderly patients among those colonised (P=0.02) with less satisfactory venous access (P=0.006) and smaller catheter bores (P=0.009). No differences were seen regarding the other evaluation criteria: gender, aetiology, site of intervention, number and site of venepuncture procedures, blood pressure and duration of catheterisation. The absolute value of colonisation seen in our study appears consistent with those reported in the literature, but no comparisons are possible due to widely divergent catheterisation times between published work (days) and our own data (minutes). Our results suggest a reassessment of current recommendations of routine changeover of these catheters, with the attendant discomfort for patients and cost in terms of time and money. We recommend a randomised study comparing catheter colonisation levels associated with routine catheter replacement with targeted replacement based on risk factors. PMID- 19285360 TI - Reduction in surgical site infection in patients treated with microbial sealant prior to coronary artery bypass graft surgery: a case-control study. AB - Surgical site infection (SSI) is a serious complication after cardiac surgery. This case-control study investigated the effect of a cyanoacrylate-based microbial skin sealant (InteguSeal) applied preoperatively on the SSI rate in patients undergoing coronary artery bypass graft (CABG) surgery. Of 676 patients who underwent CABG surgery with or without concomitant procedure(s) between March and November 2007, 545 received standard preoperative care and 131 also received pretreatment with the microbial sealant. Of these, 90 cases pretreated with microbial sealant and 90 controls were matched using established preoperative and intraoperative risk factors for SSI. Preoperative risk scores for SSI were 9.9+/ 4.3 and 9.7+/-4.0 (P=0.747) for the microbial sealant and the control group, respectively, and combined preoperative-intraoperative risk scores were 9.7+/-4.1 and 8.7+/-3.5 (P=0.080), respectively. Carotid artery disease (P=0.019), congestive heart failure (P=0.019), acute myocardial infarction (P=0.001) and emergency surgery (P=0.026) were significantly more common in the microbial sealant group. Follow-up was 100% for both groups. Superficial or deep sternal infection 30 days post surgery developed in seven patients (7.8%) in the control group compared with one patient (1.1%) in the microbial sealant group (odds ratio 7.5). In summary, the inclusion of microbial sealant in preoperative patient preparation seems to reduce the incidence of SSI following CABG surgery; further larger studies are needed before firm conclusions can be drawn. PMID- 19285361 TI - Mostly dead: can science help with disaster triage? PMID- 19285362 TI - Evaluation of cerebral oxygenation during procedural sedation in children using near infrared spectroscopy. AB - STUDY OBJECTIVE: We evaluate the utility of near infrared spectroscopy monitoring and its correlation to conventional respiratory monitors during changes in cardiorespiratory characteristics during pediatric procedural sedation. METHODS: In this prospective observational study of 100 children, cerebral oxygenation (rSO(2)), pulse oximetry (SpO(2)), and end-tidal carbon dioxide (etco(2)) were monitored continuously. Values were manually recorded at least every 3 minutes from baseline until 30 minutes after sedative administration, resulting in 1,515 triplicate (simultaneous near infrared spectroscopy/etco(2)/SpO(2)) measurements. Correlations between conventional monitoring characteristics (SpO(2) and etco(2)) and rSO(2) were determined, with focus during adverse cardiorespiratory events. RESULTS: Cerebral oxygenation remained normal in 1,483 of 1,515 measurements (97.9%). rSO(2) decreased significantly during 3 of 13 hypoxic events occurring in 13 patients and during 5 of 17 hypercarbic events occurring in 8 patients, with 15 measurements of greater than 20% decrease from baseline. Cerebral oxygenation increased transiently in 88% of children. During 31 cerebral desaturation recordings, 3 hypoxic recordings (9.3%, always in combination with hypercarbia) and 5 hypercarbic recordings (15.6%) were observed, whereas in 23 (74.2%), cardiorespiratory characteristics were unchanged. There was poor correlation between rSO(2) and both SpO(2) and etco(2), with correlation coefficients of 0.05 (95% confidence interval 0.04 to 0.07) and 0.01 (95% confidence interval -0.01 to 0.02), respectively. CONCLUSION: Cerebral oxygenation as measured by near infrared spectroscopy demonstrated few significant negative changes during pediatric procedural sedation. Transient cardiorespiratory events seldom altered rSO(2), with hypercarbia having a greater effect than hypoxemia. However, cerebral desaturations frequently occurred without associated cardiorespiratory changes. PMID- 19285363 TI - [Hospitalizations due to chronic obstructive pulmonary disease in the elderly]. AB - INTRODUCTION: Hospitalizations for decompensation of chronic obstructive pulmonary disease (COPD) mainly occur in the elderly. The aim of this study was to describe the characteristics of octogenarians admitted for COPD and to compare these characteristics with those in a younger group. MATERIAL AND METHODS: All patients hospitalized for COPD in an acute care hospital over three time periods were studied. All patients met spirometric criteria for COPD. A questionnaire evaluating items on prior admissions, days of hospital stay and readmissions in the subsequent year was completed. Data on comorbidity (the Charlson index), functional dependency (Katz index), depression (Yesavage scale), domiciliary medication, socioeconomic position, social resources, and quality of life, among other factors, were gathered. RESULTS: We studied 390 patients, with a mean age of 72 years (SD 9.6), of whom 88 (22%) were aged more than 80 years old. The mean length of hospital stay was 11.4 days, FEV(1) at discharge was 39% of the theoretical value, and 55% of the patients were readmitted in the following year, with no differences between age groups. Patients older than 80 years had a lower body mass index (P<.03), greater comorbidity (P<.001), greater functional dependency (P<.001) and worse scores on the Pffeifer (P<.001) and Yesavage scales (P<.01). CONCLUSIONS: Octogenarians hospitalized for COPD exacerbations have greater comorbidity, depressive features and functional dependency than younger patients. Nevertheless, no differences were found in the length of hospital stay or in readmissions in the following year. PMID- 19285364 TI - [Alternative to surgery in basal cell carcinoma in the elderly population: imiquimod 5% cream]. AB - Basal cell carcinoma (BCC) is the most common non-melanoma skin cancer. The incidence of this entity increases with age, peaking at the age of 70 years. This study included 97 patients aged more than 65 years old, with a diagnosis of BCC. These patients were treated with imiquimod applied three times weekly for 6 weeks. Clinical remission was 67% after 10 weeks and was 49.5% after 1 year. Imiquimod is a highly interesting therapeutic modality in the elderly, in whom surgery is often contraindicated. This regimen eradicates a high percentage of tumors and reduces local adverse effects, which are poorly tolerated by aged skin. PMID- 19285365 TI - [Use of the Cambridge Neuropsychological Test Automated Battery for the diagnosis of mild cognitive impairment. A pilot study in a Spanish sample]. AB - INTRODUCTION: Suitable assessment tools for the diagnosis of mild cognitive impairment (MCI) could facilitate the early detection of Alzheimer's disease and other types of dementia. The aim of the present study was to assess the utility of the main memory and reaction time tests of the Cambridge Neuropsychological Test Automated Battery (CANTAB) for detecting MCI. MATERIAL AND METHODS: Episodic and working memory and reaction and movement times were tested in 16 MCI patients, classified according to Petersen's criteria, and in 15 healthy individuals. RESULTS: ANOVA showed that the performance of the MCI group was significantly poorer than that of the control group in movement time and episodic memory tests, pattern recognition, delayed matching to sample and paired associates learning. Performance in these tests correlated with the measures of general cognitive performance. However, the performance of both groups was similar in simple reaction times and in the spacial working memory tests. CONCLUSIONS: The CANTAB episodic memory tests and the movement time measures are effective instruments to detect MCI. PMID- 19285366 TI - [Cutaneous leishmaniasis in rheumatoid arthritis]. AB - INTRODUCTION: Cutaneous leishmaniasis is a protozoal infection. Its prevalence is increasing, especially in immunocompromised subjects. CASE REPORTS: We report four patients with rheumatoid arthritis, treated with methotrexate and prednisone who developed cutaneous leishmaniasis. Clinical outcome was favorable after institution of antimony therapy in three cases despite the continuation of methotrexate and prednisone. One patient failed to respond to therapy. DISCUSSION: The frequency of cutaneous leishmaniasis is increasing especially in immunocompromised subjects. In our patients, rheumatoid arthritis, corticosteroid therapy and methotrexate were predisposing factors of cutaneous leishmaniasis. PMID- 19285367 TI - Transient analysis of trunk response in sudden release loading using kinematics driven finite element model. AB - BACKGROUND: Sudden trunk perturbations occur in various occupational and sport activities. Despite numerous measurement studies, no comprehensive modeling simulations have yet been attempted to investigate trunk biodynamics under sudden loading/unloading. METHODS: Dynamic kinematics-driven approach was used to evaluate the temporal variation of trunk muscle forces, internal loads and stability before and after a sudden release of a posterior horizontal load. Measured post-disturbance trunk kinematics, as input, and muscle electromyography (EMG) activities, for qualitative validation, were considered. FINDINGS: Computed agonist and antagonist muscle forces before and after release agreed well with reported EMG activities and demonstrated basic response characteristics such as activation latency and reflex activation. The trunk was found quite stable before release and in early post-release period. Larger applied load substantially increased trunk kinematics, muscle forces and spinal loads. INTERPRETATION: Excessive spinal loads due to large muscle forces in sudden loading conditions is a risk factor as the central nervous system attempts to reflexively control the sudden disturbances, a situation that further deteriorates under larger perturbations and longer latency periods. Predictions indicate the potential of the kinematics-driven model in ergonomics as well as training and rehabilitation programs. PMID- 19285368 TI - Interacting effects of sulphate pollution, sulphide toxicity and eutrophication on vegetation development in fens: a mesocosm experiment. AB - Both eutrophication and SO4 pollution can lead to higher availability of nutrients and potentially toxic compounds in wetlands. To unravel the interaction between the level of eutrophication and toxicity at species and community level, effects of SO4 were tested in nutrient-poor and nutrient-rich fen mesocosms. Biomass production of aquatic and semi-aquatic macrophytes and colonization of the water layer increased after fertilization, leading to dominance of highly competitive species. SO4 addition increased alkalinity and sulphide concentrations, leading to decomposition and additional eutrophication. SO4 pollution and concomitant sulphide production considerably reduced biomass production and colonization, but macrophytes were less vulnerable in fertilized conditions. The experiment shows that competition between species, vegetation succession and terrestrialization are not only influenced by nutrient availability, but also by toxicity, which strongly interacts with the level of eutrophication. This implies that previously neutralized toxicity effects in eutrophied fens may appear after nutrient reduction measures have been taken. PMID- 19285369 TI - Clonal differences in survival capacity, copper and zinc accumulation, and correlation with leaf polyamine levels in poplar: a large-scale field trial on heavily polluted soil. AB - Three ex situ collections of poplar clones from natural populations of Populus alba and P. nigra growing in northern Italy were assessed for their genetic dissimilarity (GD) by means of amplified fragment length polymorphism (AFLP). The high GD evidenced within populations was exploited for screening 168 clones in a field trial on heavy metal-polluted soil. After one growth season, clonal differences in plant survival and growth were observed. On the basis of performance, six clones were singled out, and used to evaluate copper and zinc accumulation in different organs. Clonal differences in metal concentrations were most evident for leaves and stems; one clone of P. alba (AL35) had a distinctly higher concentration of both metals in the roots. Leaf polyamine (putrescine, spermidine, spermine) profiles correlated with tissue metal concentrations, depending on the clone, plant organ and metal. In particular, the high metal accumulating clone AL35 exhibited a dramatically higher concentration of free and conjugated putrescine. Overall, the results indicate that, given the high GD of Populus even within populations, it is possible to identify genotypes best suited for soil clean-up, and useful also for investigating physiological markers associated with high metal accumulation/tolerance. PMID- 19285370 TI - Polychlorinated biphenyls and organochlorine pesticides in various bird species from northern China. AB - Little data are available on organochlorine contamination in Chinese terrestrial birds of prey. This study examined the presence of PCBs, DDTs and other organochlorine pesticides in various raptors from northern China. DDE exhibited the highest concentrations among targeted compounds. Greatest levels (23.5-1020 mg/kg lipid weight) were observed in Eurasian sparrowhawks. This may be due to their stopover in southeastern China, where high DDT and dicofol applications have been documented. Residential kestrels exhibited much lower DDE, but similar PCB and HCH concentrations. SigmaTEQs and PCB-126/-77 concentration ratios exhibited significant positive correlations with SigmaPCB concentrations, respectively. Similar results were also demonstrated by a meta-analysis of previously published data across avian species. Possible hepatic sequestration of coplanar PCB-77, -126, -169 and -118 was observed as liver TEQs increased in Eurasian sparrowhawks. These observations may indicate an induction of CYP1A enzymes, as a result of elevated contamination in some species. PMID- 19285371 TI - Diarrhoea, acute respiratory infection, and fever among children in the Democratic Republic of Congo. AB - Several years of war have created a humanitarian crisis in the Democratic Republic of Congo (DRC) with extensive disruption of civil society, the economy and provision of basic services including health care. Health policy and planning in the DRC are constrained by a lack of reliable and accessible population data. Thus there is currently a need for primary research to guide programme and policy development for reconstruction and to measure attainment of the Millennium Development Goals (MDGs). This study uses the 2001 Multiple Indicators Cluster Survey to disentangle children's health inequalities by mapping the impact of geographical distribution of childhood morbidity stemming from diarrhoea, acute respiratory infection, and fever. We observe a low prevalence of childhood diarrhoea, acute respiratory infection and fever in the western provinces (Kinshasa, Bas-Congo and Bandundu), and a relatively higher prevalence in the south-eastern provinces (Sud-Kivu and Katanga). However, each disease has a distinct geographical pattern of variation. Among covariate factors, child age had a significant association with disease prevalence. The risk of the three ailments increased in the first 8-10 months after birth, with a gradual improvement thereafter. The effects of socioeconomic factors vary according to the disease. Accounting for the effects of the geographical location, our analysis was able to explain a significant share of the pronounced residual geographical effects. Using large scale household survey data, we have produced for the first time spatial residual maps in the DRC and in so doing we have undertaken a comprehensive analysis of geographical variation at province level of childhood diarrhoea, acute respiratory infection, and fever prevalence. Understanding these complex relationships through disease prevalence maps can facilitate design of targeted intervention programs for reconstruction and achievement of the MDGs. PMID- 19285372 TI - Childhood drowning in Matlab, Bangladesh: an in-depth exploration of community perceptions and practices. AB - While reductions in infectious disease have resulted in impressive declines in child mortality in Bangladesh, drowning is becoming proportionately more important as a major cause of death, accounting for at least 19% of deaths of children between 1 and 4 years of age in trend analysis since 2000. Little is known about indigenous beliefs and behaviors associated with drowning, which may be critical to preventing child-related drowning deaths. Qualitative research was carried out over 13 months in Matlab, Bangladesh to describe the indigenous explanatory model of drowning and to identify behavioral factors increasing the risk for drowning deaths. Methods included cognitive mapping procedures as well as open-ended interviews with families who had lost a child or experienced a near death due to drowning and families with at least one child under 5 years living near a body of water. Along with diarrhea, fever, and pneumonia, drowning is perceived as a leading cause of child death. Causal explanations are primarily associated with "evil spirits" believed to entice young children to water or bewitch mothers so that they forget about the child. Another primary interpretation relates to a water goddess known to prey on small children. When a young child is discovered in water, parents refrain from rescuing the child due to a belief that if a parent touches a drowning child, the child will die. After the child is removed from the water, traditional practices that have no known benefit are employed. The research identified locally constructed beliefs and practices such as refraining from touching the child that may increase the incidence of drowning deaths. Future efforts are required to address these beliefs and assess the feasibility, cultural acceptability and effectiveness of strategies designed to prevent drowning. PMID- 19285373 TI - Do biological, sociodemographic, and behavioral characteristics explain racial/ethnic disparities in preterm births? AB - Many studies find racial/ethnic disparities in a diverse set of birth outcomes. However few empirical studies have examined the existence and possible explanations for racial/ethnic disparities in preterm births using a diverse set of racial/ethnic categories and a nationally representative sample of births. This research fills that gap. Using data from the US Early Childhood Longitudinal Study - Birth Cohort (ECLS-B), this research first explores the distribution of biological, sociodemographic, and behavioral characteristics of mothers and infants based on seven categories of maternal race/ethnicity. Next, multivariable logistic regression models are estimated in a nested manner to test for possible explanations for racial/ethnic disparities in preterm births. Lastly, race stratified models are estimated to better elucidate the mechanism leading to racial/ethnic disparities in preterm births. Results from the chi-square tests of significance for racial/ethnic differences indicate that all variables used in this analysis, except for infant's gender, differ significantly based on maternal race/ethnicity. Results from the full multivariable logistic regression model finds that the only racial/ethnic disparity found in preterm births is observed for infants born to Native American mothers compared to non-Hispanic white mothers, once all variables are controlled for in the model. Race-stratified models indicate that maternal health complications and prenatal care adequacy offer the most potential in explaining remaining racial/ethnic disparities in preterm births. Results from this research support the need to increase access to appropriate and timely prenatal care for women of all races/ethnicities in an effort to reduce racial/ethnic disparities in preterm births. PMID- 19285374 TI - Differences in bioregional classifications among four aquatic biotic groups: implications for conservation reserve design and monitoring programs. AB - Bioregional classifications are used extensively for conservation management and monitoring programs. This study used generalised dissimilarity modelling (GDM) to test the ability of different regional classifications of four groups of aquatic biota to be used as surrogates for each other. Classifications were derived for aquatic macrophytes, macroinvertebrates, freshwater fish and frogs using community-level modelling, or GDM, which relates the biotic assemblage structure with environmental variables. Six regions were defined for each biotic group for the State of New South Wales. Regional classifications differed markedly between the different biotic groups because the environmental drivers that were related to species turnover throughout the region differed among groups. Altitude and rainfall were the strongest drivers of species turnover among the groups. Results suggest that physiographic variables should be incorporated in reserve design and monitoring programs to explicitly address differences in classifications between similar biotic groups. PMID- 19285375 TI - Velocity variability in ultrasonic Doppler examinations. AB - Ultrasonic Doppler examination with spectral waveform has been used for the classification and surveillance of carotid artery stenosis for over 25 years. Progression/regression between examinations can be identified with 95% confidence if the velocity measurements change by more than two times the root mean square difference (RMSD) of the repeat measurement. Peak systolic velocity and end diastolic velocity measurements at a Doppler angle of 60 degrees were repeated in 47 carotid examinations. Measurement difference between sonographers and between instruments was tabulated. Root mean square difference was 11 cm/s (RMS%D = 11%) for systolic and 7 cm/s (RMS%D = 21%) for diastolic velocity measurements (excluding one severe stenosis case). Results for differences between sonographers and between instruments were similar to the overall results. In serial arterial studies using this Doppler velocimetry method, a difference exceeding 23 cm/s (21%) systolic, or 14 cm/s (42%) diastolic velocity indicates a significant (2 x RMSD) hemodynamic change. PMID- 19285376 TI - How effective are patient-based educational interventions in the management of cancer pain? Systematic review and meta-analysis. AB - This review aimed to quantify the benefit of patient-based educational interventions in the management of cancer pain. We undertook a systematic review and meta-analysis of experimentally randomised and non-randomised controlled clinical trials identified from six databases from inception to November 2007.Two reviewers independently selected trials comparing intervention (formal instruction on cancer pain and analgesia on an individual basis using any medium) to usual care or other control in adults with cancer pain. Methodological quality was assessed, and data extraction undertaken by one reviewer with a second reviewer checking for accuracy. We used random effects model to combine the effect estimates from studies. Main outcome measures were effects on knowledge and attitudes towards cancer pain and analgesia, and pain intensity. Twenty-one trials (19 randomised) totalling 3501 patients met inclusion criteria, and 15 were included in the meta-analysis. Compared to usual care or control, educational interventions improved knowledge and attitudes by half a point on 0-5 rating scale (weighted mean difference 0.52, 95% confidence interval 0.04-1.0), reduced average pain intensity by over one point on 0-10 rating scale (WMD -1.1, 1.8 to -0.41) and reduced worst pain intensity by just under one point (WMD 0.78, -1.21 to -0.35). We found equivocal evidence for the effect of education on self-efficacy, but no significant benefit on medication adherence or on reducing interference with daily activities. Patient-based educational interventions can result in modest but significant benefits in the management of cancer pain, and are probably underused alongside more traditional analgesic approaches. PMID- 19285377 TI - Step aside tobacco, chlorine could be man's next great carcinogen. PMID- 19285378 TI - A two-host fosmid system for functional screening of (meta)genomic libraries from extreme thermophiles. AB - A new cloning system is described, which allows the construction of large-insert fosmid libraries in Escherichia coli and the transfer of the recombinant libraries to the extreme thermophile Thermus thermophilus via natural transformation. Libraries are established in the thermophilic host by site specific chromosomal insertion of the recombinant fosmids via single crossover or double crossover recombination at the T. thermophilus pyr locus. Comparative screening of a fosmid library constructed from genomic DNA from the thermophilic spirochaete, Spirochaeta thermophila, for clones expressing thermoactive xylanase activity revealed that 50% of the fosmids that conferred xylanase activity upon the corresponding T. thermophilus transformants did not give rise to xylanase positive E. coli clones, indicating that significantly more S. thermophila genes are functionally expressed in T. thermophilus than in E. coli. The novel T. thermophilus host/vector system may be of value for the construction and functional screening of recombinant DNA libraries from individual thermophilic or extremely thermophilic organisms as well as from complex metagenomes isolated from thermophilic microbial communities. PMID- 19285379 TI - Hip flexion after Birmingham hip resurfacing: role of cup anteversion, anterior femoral head-neck offset, and head-neck ratio. AB - We aimed to investigate the factors affecting range of flexion after hip resurfacing. A total of 82 cases, operated by a single surgeon, were assessed at a mean of 43 months. The Einzel-Bild-Roentgen-Analysis for the acetabular cup software was used to measure socket orientation. Range of flexion had a moderate positive correlation with cup anteversion (R = 0.26, P = .017), weak but significant negative correlation with neck diameter (R = -0.23, P = .042), and none with anterior femoral head-neck offset. Using multivariate analysis that adjusted for age, sex, cup anteversion and inclination, head-neck offset ratio, head-neck ratio, and neck diameter, the only significant correlate of flexion was cup anteversion (P = .017). Care should be taken during cup placement to allow adequate anteversion to be maintained in Birmingham hip resurfacing because this can affect flexion range of motion. PMID- 19285380 TI - Management of periprosthetic femur fractures with severe bone loss using impaction bone grafting technique. AB - We present a technique of femoral impaction grafting used for the treatment of periprosthetic femur fractures with severe bone loss after total hip arthroplasty. Seven patients with femoral fractures with compromise of the femoral isthmus were treated with femoral component revision using the impaction grafting bone technique. The average age was 64 years (range, 44-72 years), and 2 patients required mesh augmentation at the time of surgery. The average follow-up for this group of patients was 56 months (range, 39-92 months). Radiographic evaluation revealed healed fractures in all patients and no evidence of implant loosening at a mean of 56 months (range, 39-92 months). There were no cases of infections or dislocations in this series. Impaction grafting technique can be useful in the treatment of periprosthetic femur fractures when bone loss and canal geometry preclude the use of fully coated femoral components. PMID- 19285381 TI - Altered lateralisation of emotional prosody processing in schizophrenia. AB - Alterations of cerebral lateralisation in schizophrenia have been reported consistently, and a reduced left-lateralisation has been suggested for language functions. Speech contains non-verbal information, e.g. prosody, and on a behavioural level, the extraction of emotional information from prosody is often impaired in schizophrenia. A previous functional magnetic resonance imaging study suggests increased left-lateralisation in schizophrenia during prosody processing, but did not disentangle effects of speech processing as such and emotional prosody processing. Here, we used meaningless syllables spoken with neutral, angry or fearful speech melody and measured blood oxygen level-dependent (BOLD) responses in 15 in-patients with schizophrenia and 15 healthy control participants matched for age and gender. Lateralisation indices were calculated for responses to emotional versus neutral prosody, and for all types of prosody versus baseline. Compared to control participants, patients with schizophrenia showed an increased right-lateralisation of emotional and non-emotional prosody processing in the temporal and parietal cortex. This right-lateralisation was increased in patients with reduced right-handedness and decreased in patients with stronger negative symptoms, particularly affective blunting, and with longer hospitalisation. Although patients with schizophrenia performed worse in emotion identification, this deficit was not related to lateralisation indices. Enhanced right-lateralisation to prosody resembles previous findings on laterality changes in speech processing and might suggest a common underlying cause in the organization of language functions. PMID- 19285382 TI - Structure and seasonal dynamics of the protozoan community (heterotrophic flagellates, ciliates, amoeboid protozoa) in the plankton of a large river (River Danube, Hungary). AB - Seasonal dynamics of all major protozoan groups were investigated in the plankton of the River Danube, upstream of Budapest (Hungary), by bi-weekly sampling over a 1-year long period. Sixty-one heterotrophic flagellate, 14 naked amoeba, 50 testate amoeba, 4 heliozoan and 83 ciliate morphospecies were identified. The estimated abundance ranges of major groups throughout the year were as follows: heterotrophic flagellates, 0.27-7.8 x 10(6)ind.l(-1); naked amoebae, max. 3300ind.l(-1); testaceans, max. 1600ind.l(-1); heliozoans, max. 8500ind.l(-1); ciliates, 132-34,000ind.l(-1). In terms of biovolume, heterotrophic flagellates dominated throughout the year (max. 0.58mm(3)l(-1)), and ciliates only exceeded their biovolume in summer (max. 0.76mm(3)l(-1)). Naked amoeba and heliozoan biovolume was about one, and testacean biovolume 1-3, orders of magnitude lower than that of ciliates. In winter, flagellates, mainly chrysomonads, had the highest biomass, whilst ciliates were dominated by peritrichs. In 2005 from April to July a long spring/summer peak occurred for all protozoan groups. Beside chrysomonads typical flagellates were choanoflagellates, bicosoecids and abundant microflagellates (large chrysomonads and Collodictyon). Most abundant ciliates were oligotrichs, while Phascolodon, Urotricha, Vorticella, haptorids, Suctoria, Climacostomum and Stokesia also contributed significantly to biovolume during rapid succession processes. In October and November a second high protozoan peak occurred, with flagellate dominance, and slightly different taxonomic composition. PMID- 19285383 TI - Leaching of heavy metals from E-waste in simulated landfill columns. AB - In recent history the volume of electronic products purchased by consumers has dramatically escalated. As a result this has produced an ever-increasing electronic waste (E-waste) stream, which has generated concerns regarding the E waste's potential for adversely impacting the environment. The leaching of toxic substances from obsolete personal computers (PCs) and cathode ray tubes (CRTs) of televisions and monitors, which are the most significant components in E-waste stream, was studied using landfill simulation in columns. Five columns were employed. One column served as a control which was filled with municipal solid waste (MSW), two columns were filled with a mixture of MSW and CRTs, and the other two were filled with MSW and computer components including printed wire boards, hard disc drives, floppy disc drives, CD/DVD drives, and power supply units. The leachate generated from the columns was monitored for toxic materials throughout the two-year duration of the study. Results indicate that lead (Pb) and various other heavy metals that were of environmental and health concern were not detected in the leachate from the simulators. When the samples of the solids were collected from underneath the E-waste in the columns and were analyzed, significant amount of Pb was detected. This indicates that Pb could readily leach from the E-waste, but was absorbed by the solids around the E-waste materials. While Pb was not observed in the leachate in this study, it is likely that the Pb would eventually enter the leachate after a long term transport. PMID- 19285384 TI - Sustainable solutions for solid waste management in Southeast Asian countries. AB - Human activities generate waste and the amounts tend to increase as the demand for quality of life increases. Today's rate in the Southeast Asian Nations (ASEANs) is alarming, posing a challenge to governments regarding environmental pollution in the recent years. The expectation is that eventually waste treatment and waste prevention approaches will develop towards sustainable waste management solutions. This expectation is for instance reflected in the term 'zero emission systems'. The concept of zero emissions can be applied successfully with today's technical possibilities in the agro-based processing industry. First, the state of-the-art of waste management in Southeast Asian countries will be outlined in this paper, followed by waste generation rates, sources, and composition, as well as future trends of waste. Further on, solutions for solid waste management will be reviewed in the discussions of sustainable waste management. The paper emphasizes the concept of waste prevention through utilization of all wastes as process inputs, leading to the possibility of creating an ecosystem in a loop of materials. Also, a case study, focusing on the citrus processing industry, is displayed to illustrate the application of the aggregated material input-output model in a widespread processing industry in ASEAN. The model can be shown as a closed cluster, which permits an identification of opportunities for reducing environmental impacts at the process level in the food processing industry. Throughout the discussion in this paper, the utilization of renewable energy and economic aspects are considered to adapt to environmental and economic issues and the aim of eco-efficiency. Additionally, the opportunities and constraints of waste management will be discussed. PMID- 19285385 TI - The use of PTC and RFA as treatment alternatives with low procedural morbidity in non-small cell lung cancer. AB - Minimally invasive percutaneous ablative therapies for treating lung cancers are currently being studied as treatment alternatives. This present study investigated the efficacies of percutaneous thoracic cryotherapy (PTC) and radiofrequency ablation (RFA) on clinical courses of pulmonary malignant tumours, especially in the setting of non-surgical candidates. Sixty-five patients with lung malignancy underwent sixty-seven sessions of RFA and nine sessions of PTC. We evaluated the results of RFA and PTC including efficacies, local progression rate, survival rate, and complications. Twenty-nine patients (43.3%) treated with RFA and six patients (66.7%) with PTC attained complete ablation. In small-sized lung mass (3 cm), complete ablation rate of RFA and PTC was increased to 76.2% and 85.7%, respectively. Additionally, we have found that the complete ablation group had significantly higher survival duration and progression free survival duration compared with the partial ablation group. Moreover, the complication profile was acceptable and the pain associated with the procedures disappeared within 1 day; 42 patients (62.7%) after RFA and all patients after PTC. This study provides evidence for the use of PTC and RFA as treatment alternatives with low procedural morbidity in the management of inoperable pulmonary malignant tumours, although the current study is limited by the small sample size and the short follow-up period. PMID- 19285386 TI - Adjuvant therapy after radical prostatectomy: clinical considerations. AB - There are no standardised guidelines for the role of adjuvant radiotherapy post prostatectomy. Evidence that may shed light into its appropriate usage is marred by the difficulty in defining adjuvant radiotherapy and failure of some studies to adequately distinguish it from salvage radiotherapy. The ability to identify which prostate cancers are likely to benefit from immediate post-operative radiotherapy would enable the judicious use of this modality and minimise the unnecessary exposure to its side-effects. This review outlines the pre- and post surgical clinical considerations that form the discussion around the requirement for adjuvant therapy; radiotherapy, androgen deprivation therapy and chemotherapy. Principles for the use of local and systemic therapies are discussed. PMID- 19285387 TI - Production, partial purification and characterization of organic solvent tolerant lipase from Burkholderia multivorans V2 and its application for ester synthesis. AB - Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solvent tolerant lipase (6.477 U/mL). This lipase exhibited maximum stability in n-hexane retaining about 97.8% activity for 24h. After performing statistical optimization of medium components for lipase production, a 2.2-fold (14 U/mL) enhancement in the lipase production was observed. The crude lipase from BMV2 was partially purified by ultrafiltration and gel permeation chromatography with 24.64-fold purification. The K(m) and V(max) values for partially purified BMV2 lipase were found to be 1.56 mM and 5.62 micromoles/mg min. The metal ions Ca(2+), Mg(2+) and Mn(2+) had stimulatory effect on lipase activity, whereas Cu(2+), Fe(2+) and Zn(2+) strongly inhibited the lipase activity. EDTA and PMSF at 10mM concentration strongly inhibited the lipase activity. Non-ionic and anionic surfactants stimulated the lipase activity. BMV2 lipase was proved to be efficient in synthesis of ethyl butyrate ester under non aqueous environment. PMID- 19285388 TI - Design of novel quinazoline derivatives and related analogues as potent and selective ALK5 inhibitors. AB - Starting from quinazoline 3a, we designed potent and selective ALK5 inhibitors over p38MAP kinase from a rational drug design approach based on co-crystal structures in the human ALK5 kinase domain. The quinazoline 3d exhibited also in vivo activity in an acute rat model of DMN-induced liver fibrosis when administered orally at 5mg/kg (bid). PMID- 19285389 TI - Tricyclic HIV integrase inhibitors V. SAR studies on the benzyl moiety. AB - SAR studies on the para-fluorobenzyl moiety of tricyclic HIV integrase inhibitors are discussed and lead compounds with potency and PK properties comparable to raltegravir were identified. PMID- 19285390 TI - Inhibitors of hepatitis C virus NS3/4A: alpha-ketoamide based macrocyclic inhibitors. AB - A novel series of hepatitis C virus (HCV) NS3/4A protease inhibitors bearing a P2 P4 macrocycle and a P1-P1' alpha-ketoamide serine trap is reported. The NS3 protease, which is essential for viral replication, is considered one of the most attractive targets for developing novel anti-HCV therapies. The optimization of both the macrocycle and the warhead portions led to the discovery of compounds 8b and 8 g with a good activity both in the enzyme as well as in the cell based (replicon) assays with favorable PK profile in a preclinical species. PMID- 19285391 TI - Synthesis, cytotoxicity, and haemolytic activity of chacotrioside lupane-type neosaponins and their germanicane-type rearrangement products. AB - The concise synthesis, via a stepwise glycosylation approach, of lupeol, betulin and betulinic acid O-glycosides bearing a chacotriosyl moiety at the C-3 position is described. All neosaponins as well as their rearrangement products of the germanicane-type were evaluated in vitro for their anticancer and haemolytic activities. Although betulinic acid and betulin 3beta-O-chacotriosides were neither cytotoxic nor haemolytic, their rearrangement products allobetulin and 28 oxoallobetulin 3beta-O-chacotriosides (9 and 10) exhibited a cytotoxicity profile up to fourfold superior to betulinic acid against human breast (MCF7) and prostate (PC-3) adenocarcinomas cell lines (IC(50)=10-18 microM). One important result was that only chacotriosides featuring non-polar functions at the C-28 position (6, 9 and 10) exerted a haemolytic activity against red blood cells. PMID- 19285392 TI - Discovery of substituted 4-anilino-2-arylpyrimidines as a new series of apoptosis inducers using a cell- and caspase-based high throughput screening assay. 2. Structure-activity relationships of the 2-aryl group. AB - As a continuation of our efforts to discover and develop the apoptosis inducing 4 anilino-2-(2-pyridyl)pyrimidines as potential anticancer agents, we explored replacing the 2-pyridyl group by other aryl groups. SAR studies showed that the 2 pyridyl group can be replaced by a 3-pyridyl, 4-pyridyl and 2-pyrazinyl group, and that the SAR for the anilino group was similar to that of the 2-pyridyl series. However, replacement of the 2-pyridyl group by a phenyl group, a 3,5 dichloro-4-pyridyl group, or a saturated ring led to inactive compounds. Several potent compounds, including 2f, 3d, 3j and 4a, with EC(50) values of 0.048-0.024 microM in the apoptosis induction assay against T47D cells, were identified through the SAR studies. In a tubulin polymerization assay, compound 2f, which was active against all the three cell lines tested (T47D, HTC116 and SNU398), inhibited tubulin polymerization with an IC(50) value of 0.5 microM, while compound 2a, which was active against T47D cells but not active against HTC116 and SNU398 cells, was not active in the tubulin assay at up to 50 microM. PMID- 19285393 TI - Discovery of 5-pyrrolopyridinyl-2-thiophenecarboxamides as potent AKT kinase inhibitors. AB - A pyrrolopyridinyl thiophene carboxamide 7 was discovered as a tractable starting point for a lead optimization effort in an AKT kinase inhibition program. SAR studies aided by a co-crystal structure of 7 in AKT2 led to the identification of AKT inhibitors with subnanomolar potency. Representative compounds showed antiproliferative activity as well as inhibition of phosphorylation of the downstream target GSK3beta. PMID- 19285394 TI - Synthesis and beta-amyloid binding properties of rhenium 2-phenylbenzothiazoles. AB - As a first step toward the development of (99m)Tc PiB analogs, we have synthesized six neutral Re 2-phenylbenzothiazoles via pendant or integrated approach. These Re compounds bind to Abeta(1-40) fibrils with fairly good affinities (K(i)=10.0-88.6nM) and have moderate lipophilicities (logP(C18)=1.21 3.26). The Re compounds prepared via the integrated approach are smaller in size, and therefore their corresponding (99m)Tc analogs would have a greater chance of crossing the blood-brain barrier well. For potential clinical applications, further optimization on the structure-activity relationship to obtain Re 2 phenylbenzothiazoles with higher binding affinities (<10nM) might be needed. The integrated approach reported here to obtain neutral, compact and lipophilic Re 2 phenylbenzothiazoles could to be applied to other high affinity pharmacophores as well as to generate (99m)Tc analogs that could hold promise for extending the use of Abeta imaging in living human brain to many more clinical settings because they could be used with SPECT. PMID- 19285395 TI - N-Hydroxy-(4-oxime)-cinnamide: a versatile scaffold for the synthesis of novel histone deacetylase [correction of deacetilase] (HDAC) inhibitors. AB - With the aim to discover novel HDAC inhibitors with high potency and good safety profiles, we have designed a small library based on a N-hydroxy-(4-oxime) cinnamide scaffold. We describe the synthesis of these novel compounds and some preliminary in vitro cytotoxic activity on three tumor cell lines, NB4, H460 and HCT116, as well as their inhibitory activity against class I, II and IV HDAC. Several 4-oxime derivatives demonstrated a promising inhibitory activity on HDAC6 and HDAC8 coupled to a good selectivity profile. PMID- 19285396 TI - Predicting the status of axillary lymph nodes in breast cancer: a multiparameter approach including axillary ultrasound scanning. AB - In a prospective study, we attempted to predict axillary metastases in 135 breast cancer patients by a preoperative multiparameter evaluation including axillary ultrasound scanning (US). After surgery, factors associated with lymph node metastases by univariate analysis were included in a multivariate model. By multivariate analysis, the stronger independent predictors of lymph node metastases were suspicious axillary US (p<0.001), tumor location in the outer quadrants (p=0.001) and high Ki-67 index (>10%) (p=0.002). A predictive model based on these variables, identified a high-risk group (20.0%) represented by women with suspicious axillary US, tumor in the outer quadrants and high Ki-67 index, with axillary metastases in 100%, whereas all patients with opposite features (8.1%) had uninvolved axillary lymph nodes. This multiparameter evaluation including axillary US may be used to optimize the selection of breast cancer patients candidate to sentinel lymph node biopsy or axillary lymph node dissection. The accuracy of this predictive model still requires prospective validation in a larger sample of women. PMID- 19285397 TI - Polymorphic members of the lag gene family mediate kin discrimination in Dictyostelium. AB - Self and kin discrimination are observed in most kingdoms of life and are mediated by highly polymorphic plasma membrane proteins. Sequence polymorphism, which is essential for effective recognition, is maintained by balancing selection. Dictyostelium discoideum are social amoebas that propagate as unicellular organisms but aggregate upon starvation and form fruiting bodies with viable spores and dead stalk cells. Aggregative development exposes Dictyostelium to the perils of chimerism, including cheating, which raises questions about how the victims survive in nature and how social cooperation persists. Dictyostelids can minimize the cost of chimerism by preferential cooperation with kin, but the mechanisms of kin discrimination are largely unknown. Dictyostelium lag genes encode transmembrane proteins with multiple immunoglobulin (Ig) repeats that participate in cell adhesion and signaling. Here, we describe their role in kin discrimination. We show that lagB1 and lagC1 are highly polymorphic in natural populations and that their sequence dissimilarity correlates well with wild strain segregation. Deleting lagB1 and lagC1 results in strain segregation in chimeras with wild-type cells, whereas elimination of the nearly invariant homolog lagD1 has no such consequences. These findings reveal an early evolutionary origin of kin discrimination and provide insight into the mechanism of social recognition and immunity. PMID- 19285398 TI - Irc15 Is a microtubule-associated protein that regulates microtubule dynamics in Saccharomyces cerevisiae. AB - Microtubules are polymers composed of alpha-beta tubulin heterodimers that assemble into microtubules. Microtubules are dynamic structures that have periods of both growth and shrinkage by addition and removal of subunits from the polymer. Microtubules stochastically switch between periods of growth and shrinkage, termed dynamic instability. Dynamic instability is coupled to the GTPase activity of the beta-tubulin subunit of the tubulin heterodimer. Microtubule dynamics are regulated by microtubule-associated proteins (MAPs) that interact with microtubules to regulate dynamic instability. MAPs in budding yeast have been identified that bind microtubule ends (Bim1), that stabilize microtubule structures (Stu2), that bundle microtubules by forming cross-bridges (Ase1), and that interact with microtubules at the kinetochore (Cin8, Kar3, Kip3). IRC15 was previously identified in four different genetic screens for mutants affecting chromosome transmission or repair [11-14]. Here we present evidence that Irc15 is a microtubule-associated protein, localizing to microtubules in vivo and binding to purified microtubules in vitro. Irc15 regulates microtubule dynamics in vivo and loss of IRC15 function leads to delayed mitotic progression, resulting from failure to establish tension between sister kinetochores. PMID- 19285399 TI - Reduced effectiveness of selection caused by a lack of recombination. AB - Genetic recombination associated with sexual reproduction is expected to have important consequences for the effectiveness of natural selection. These effects may be evident within genomes, in the form of contrasting patterns of molecular variation and evolution in regions with different levels of recombination. Previous work reveals patterns that are consistent with a benefit of recombination for adaptation at the level of protein sequence: both positive selection for adaptive variants and purifying selection against deleterious ones appear to be compromised in regions of low recombination [1-11]. Here, we re examine these patterns by using polymorphism and divergence data from the Drosophila dot chromosome, which has a long history of reduced recombination. To avoid confounding selection and demographic effects, we collected these data from a species with an apparently stable demographic history, Drosophila americana. We find that D. americana dot loci show several signatures of ineffective purifying and positive selection, including an increase in the rate of protein evolution, an increase in protein polymorphism, and a reduction in the proportion of amino acid substitutions attributable to positive selection. PMID- 19285400 TI - Decoding neuronal ensembles in the human hippocampus. AB - BACKGROUND: The hippocampus underpins our ability to navigate, to form and recollect memories, and to imagine future experiences. How activity across millions of hippocampal neurons supports these functions is a fundamental question in neuroscience, wherein the size, sparseness, and organization of the hippocampal neural code are debated. RESULTS: Here, by using multivariate pattern classification and high spatial resolution functional MRI, we decoded activity across the population of neurons in the human medial temporal lobe while participants navigated in a virtual reality environment. Remarkably, we could accurately predict the position of an individual within this environment solely from the pattern of activity in his hippocampus even when visual input and task were held constant. Moreover, we observed a dissociation between responses in the hippocampus and parahippocampal gyrus, suggesting that they play differing roles in navigation. CONCLUSIONS: These results show that highly abstracted representations of space are expressed in the human hippocampus. Furthermore, our findings have implications for understanding the hippocampal population code and suggest that, contrary to current consensus, neuronal ensembles representing place memories must be large and have an anisotropic structure. PMID- 19285401 TI - Reduced neural integration of letters and speech sounds links phonological and reading deficits in adult dyslexia. AB - Developmental dyslexia is a specific reading and spelling deficit affecting 4% to 10% of the population. Advances in understanding its origin support a core deficit in phonological processing characterized by difficulties in segmenting spoken words into their minimally discernable speech segments (speech sounds, or phonemes) and underactivation of left superior temporal cortex. A suggested but unproven hypothesis is that this phonological deficit impairs the ability to map speech sounds onto their homologous visual letters, which in turn prevents the attainment of fluent reading levels. The present functional magnetic resonance imaging (fMRI) study investigated the neural processing of letters and speech sounds in unisensory (visual, auditory) and multisensory (audiovisual congruent, audiovisual incongruent) conditions as a function of reading ability. Our data reveal that adult dyslexic readers underactivate superior temporal cortex for the integration of letters and speech sounds. This reduced audiovisual integration is directly associated with a more fundamental deficit in auditory processing of speech sounds, which in turn predicts performance on phonological tasks. The data provide a neurofunctional account of developmental dyslexia, in which phonological processing deficits are linked to reading failure through a deficit in neural integration of letters and speech sounds. PMID- 19285402 TI - Hermaphroditic sex allocation evolves when mating opportunities change. AB - The optimal deployment of reproductive resources by hermaphrodites to male versus female function (i.e., their sex allocation) depends directly on opportunities for mating. If hermaphrodites occur among females, selection should favor those with a male-biased allocation because increased male allocation enhances siring success when eggs are abundant. Similarly, when hermaphrodites co-occur with males, selection should favor those that bias their allocation toward their female function. We tested these predictions by allowing hermaphrodites of the plant Mercurialis annua to evolve in either the presence or absence of males. In the presence of males, hermaphrodites did not evolve, probably because they were already strongly female biased in the base population. However, hermaphrodites mating in the absence of males evolved greater male allocation, as predicted. Our results provide the first demonstration of an evolutionary response to the frequency of unisexuals in hermaphroditic sex allocation, and they verify the quantitative phase predicted by models for the transition between hermaphroditism and dioecy. PMID- 19285403 TI - Factors affecting the diversity of DNA replication licensing control in eukaryotes. AB - Replication of eukaryotic genomes is limited to once per cell cycle, by a two step mechanism. DNA replication origins are first "licensed" during G1 phase by loading of an inactive DNA helicase (Mcm2-7) into pre-replicative complexes (pre RCs). Initiation then occurs during S phase, triggered by cyclin-dependent kinases (CDKs), which promote recruitment of proteins required for helicase activation and replisome assembly. CDKs and the anaphase promoting complex/cyclosome (APC/C) restrict licensing to G1 phase by directly and indirectly regulating pre-RC components, including ORC, Cdc6, Cdt1, and Mcm2-7. Despite the fundamental importance of licensing regulation, the mechanisms by which pre-RC components are regulated differ widely across Eukarya. Here we show that even within the genus Saccharomyces, Cdc6 is regulated differently in different species. We propose that two factors contribute to the rapid evolution of licensing regulation. The first is redundancy: eliminating any single pre-RC regulatory mechanism has very little affect on viability. The second is interchangeability: we show that regulatory mechanisms can be swapped between pre RC components without compromising the block to re-replication. These experiments provide a framework for understanding the diversity of licensing regulation in eukaryotes and provide new tools for manipulating the chromosome-replication cycle. PMID- 19285404 TI - Signal jamming mediates sexual conflict in a duetting bird. AB - Signal evolution in social animals has produced a wide variety of communal displays, many of them remarkable feats of complex coordination. The two main explanations for this temporal precision are: (1) it evolves as a cooperative signal of coalition quality or (2) it minimizes signal jamming (i.e., interference of one signal by another). However, support for the first hypothesis is inconclusive, and the role of jamming in communal signaling strategies remains unknown. Here, we use playback experiments to examine how social context influences the structure of duets in a pair-living antbird (Hypocnemis peruviana). The results show that, although resident pairs produced coordinated duets when responding to rival pairs, conflicts of interest caused duet coordination to break down. Specifically, females responded to unpaired sexual rivals by jamming the signals of their own mates, who in turn compensated by adjusting their signals to avoid interference. In demonstrating this interaction, we provide the first evidence that signal jamming occurs between mates and that strategies for reducing jamming can result in increased signal complexity. These findings highlight the importance of jamming avoidance in determining the structure of duets and suggest that conflict between signalers, rather than cooperation alone, may drive the evolution of sophisticated communal displays in social animals. PMID- 19285405 TI - Development of global motion perception requires early postnatal exposure to patterned light. AB - The accurate representation of the motion of external objects is one of the more important tasks of the visual areas of the brain because motion by itself can provide sufficient information for discriminating visual forms and hence breaking camouflage. Whereas the analysis of the motion of single small elements can occur in primary visual cortex (V1), the perception of a common direction of global motion of some visual elements among many is supported by extrastriate cortical areas [1, 2]. Humans treated for binocular congenital cataracts afterward exhibit extreme deficits of global motion, but after monocular cataracts, the deficits are minimal [3]. These observations suggest a need for normal early patterned visual experience through at least one eye for global motion perception to develop in a typical fashion. We investigated this role for early experience and its timing on kittens that were deprived of light or patterned light at different ages. Such deprivation in the first 6 weeks resulted in long-lasting (>2 yr) profound deficits of perception of global motion but no apparent effects on the perception of simple unidirectional motion. Contrary to current opinion, sensitive periods to visual deprivation in primary and extrastriate cortex may be of similar duration. PMID- 19285406 TI - Split-thickness grafting directly on visceral surfaces to protect internal organs and stabilize abdominal wall. PMID- 19285407 TI - Clinicopathologic correlates of giant pituitary adenomas. AB - Giant adenomas comprise a clinical/therapeutic subset of pituitary adenomas that pose a surgical challenge. The study population consisted of 28 patients who had giant pituitary adenomas, which are defined as tumors with a diameter greater than 5cm. Clinically, five tumors (18%) were endocrinologically functional and 23 (82%) were not. During surgery, one tumor was radically excised, four were subtotally excised, 12 were partially excised, and 11 were biopsied. All of the tumors showed typical histological features of pituitary adenoma. Of the 23 clinically non-functional adenomas, 18 were gonadotrophic tumors, four were null cell adenomas and one was a silent corticotroph adenoma. The MIB-1 labeling indices ranged from 0.1% to 2.0%. The mean topoisomerase labeling index was 0.75%. Microvessel density ranged from 0.42% to 5.55%, and there was moderately intense immunostaining for vascular endothelial growth factor. The present study found giant adenomas to be invasive but slow growing, histologically benign and often gonadotrophic in subtype. PMID- 19285408 TI - Bilateral trigeminal neuralgia associated with limited systemic sclerosis. AB - A young patient with limited systemic sclerosis and medically resistant bilateral trigeminal neuralgia was successfully treated with microvascular decompression. To our knowledge, this is the first published report of this type of case. PMID- 19285409 TI - The influence of anemia on clinical presentation and outcome of patients with first-ever atherosclerosis-related ischemic stroke. AB - There is considerable debate regarding whether anemia qualifies as a prognostic factor for stroke. The purpose of this study was twofold: first, to assess the influence of anemia on vascular risk factors and clinical presentations in patients with first-ever atherosclerosis-related ischemic stroke and, second, to evaluate whether anemia may be of prognostic importance. A total of 774 consecutive patients with first-ever atherosclerosis-related ischemic stroke were prospectively investigated. Vascular risk factors, clinical presentations and outcomes were recorded and compared between those patients with and without anemia. Stroke recurrence and mortality were recorded at the 3-year follow-up. Of the study population, 168 (21.7%) were anemic. Multivariate analysis revealed that patients with anemia were more likely to be older than 70 years (p<0.001) and have chronic renal insufficiency (p<0.001). After a mean follow-up period of 958 days, 21 (12.5%) and 24 (4.0%) of the patients in the anemic and control groups, respectively, died. Within 3 years of initial onset, the mortality rate was significantly higher in patients with anemia (p=0.021). The Kaplan-Meier analysis for patients with and without anemia showed different survival curves (Log-rank test p<0.001). Within 3 years of the onset of first-ever atherosclerosis-related ischemic stroke, patients who had anemia at the time of the initial admission had an associated higher mortality rate. The stroke risk factors of being older than 70 years and having chronic renal insufficiency were more frequently observed in those patients with anemia. PMID- 19285410 TI - Diffuse axonal injury: novel insights into detection and treatment. AB - Diffuse axonal injury (DAI) is one of the most common and important pathologic features of traumatic brain injury. The definitive diagnosis of DAI, especially in its early stage, is difficult. In addition, most therapeutic agents for patients with DAI are non-specific. The CT scan is widely used to identify signs of DAI. Although its sensitivity is limited to moderate to severe DAI, it remains a useful first-line imaging tool that may also identify co-morbid injuries such as intracerebral hemorrhage. Recently, investigations have sought to apply advanced imaging techniques and laboratory techniques to detect DAI. Meanwhile, some potential specific treatments that may protect injured axons or stimulate axonal regeneration have been developed. We review some new diagnostic technologies and specific therapeutic strategies for DAI. PMID- 19285411 TI - Gender differences in onset timing and activation of the muscles of the dominant knee during stair climbing. AB - An electromyographical analysis (EMG) of knee muscle activation patterns and time delays has rarely been discussed in the literature. The purpose of this study was to compare the activation time and EMG amplitude of the dominant vastus medialis and medial hamstring muscles during stair climbing. Fifteen male and 18 female subjects participated in this study. The subjects were asked to ascend and descend 14 steps, five times. There was a significant time delay difference between genders during stair climbing (F=8.37, p=0.008). The female subjects demonstrated longer time delays while descending the steps. In addition, the female subjects demonstrated significantly lower normalized EMG amplitude during down stair climbing (F=5.77, p=0.025) while the male subjects demonstrated higher normalized EMG amplitude for the vastus medialis muscle while descending the steps. These results suggest that female subjects possess an increased risk factor for knee injuries during down stair climbing due to muscle activation delays with decreased vastus medialis muscle activity. PMID- 19285412 TI - The effect of individual N-glycans on enzyme activity. AB - In a series of investigations, N-glycosylation has proven to be a key determinant of enzyme secretion, activity, binding affinity and substrate specificity, enabling a protein to fine-tune its activity. In the majority of cases elimination of all putative N-glycosylation sites of an enzyme results in significantly reduced protein secretion levels, while removal of individual N glycosylation sites often leads to the expression of active enzymes showing markedly reduced catalytic activity, with the decreased activity often commensurate with the number of glycosylation sites available, and the fully deglycosylated enzymes showing only minimal activity relative to their glycosylated counterparts. On the other hand, several cases have also recently emerged where deglycosylation of an enzyme results in significantly increased catalytic activity, binding affinity and altered substrate specificity, highlighting the very unique and diverse roles that individual N-glycans play in regulating enzyme function. PMID- 19285413 TI - Characteristic of neuraminidase inhibitory xanthones from Cudrania tricuspidata. AB - Natural polyphenolic compounds generally transpire to show relatively low inhibition against glycosidase including neuraminidase. In addition the inhibition modes of such compounds are rarely competitive. In this manuscript, a series of xanthone derivatives from Cudrania tricuspidata are shown to display nanomolar inhibitor activity against neuraminidase (EC 3.2.1.18) as well as competitive inhibition modes. Compound 8 bearing vicinal dihydroxy group on the A ring displays nanomolar activity (IC(50)=0.08+/-0.01 microM), a 200-fold increase in activity relative to that of the first reported xanthone-derived neuraminidase inhibitor, mangiferin (IC(50)=16.2+/-4.2 microM). The 6,7-vicinal dihydroxy group plays a crucial role for inhibitory activity because compound 4, which has one of these hydroxyl groups prenylated was inactive (33% at 200 microM), whereas other compounds (1-3 and 6-8) showed nanomolar activity (0.08-0.27 microM) and competitive inhibition modes. Interestingly all inhibitors manifested enzyme isomerization inhibition against neuraminidase. The most potent inhibitor, compound 8 showed similar interaction with a transition-state analogue of neuraminic acid in active site. PMID- 19285414 TI - Design, synthesis, biological evaluation and molecular modelling studies of novel quinoline derivatives against Mycobacterium tuberculosis. AB - We herein describe the synthesis and antimycobacterial activity of a series of 27 different derivatives of 3-benzyl-6-bromo-2-methoxy-quinolines and amides of 2 [(6-bromo-2-methoxy-quinolin-3-yl)-phenyl-methyl]-malonic acid monomethyl ester. The antimycobacterial activity of these compounds was evaluated in vitro against Mycobacterium tuberculosis H37Rv for nine consecutive days upon a fixed concentration (6.25 microg/mL) at day one in Bactec assay and compared to untreated TB cell culture as well as one with isoniazide treated counterpart, under identical experimental conditions. The compounds 3, 8, 17 and 18 have shown 92-100% growth inhibition of mycobacterial activity, with minimum inhibitory concentration (MIC) of 6.25 microg/mL. Based on our molecular modelling and docking studies on well-known diarylquinoline antitubercular drug R207910, the presence of phenyl, naphthyl and halogen moieties seem critical. Comparison of docking studies on different stereoisomers of R207910 as well as compounds from our data set, suggests importance of electrostatic interactions. Further structural analysis of docking studies on our compounds suggests attractive starting point to find new lead compounds with potential improvements. PMID- 19285415 TI - Solution studies on the complex of 4'-epiadriamycin-d-(CGATCG)2 followed by time resolved fluorescence measurement, diffusion ordered spectroscopy and restrained molecular dynamics simulations. AB - 4'-Epiadriamycin is a better-tolerated anthracycline drug, due to lesser cardiotoxicity. We report here a study of the 2:1 complex of 4'-epiadriamycin-d (CGATCG)(2) by proton Nuclear Magnetic Resonance Spectroscopy which show the absence of sequential connectivities between C1pG2 and C5pG6 base pair steps and presence of intermolecular cross peaks of the drug and DNA. Our studies establish the role of 9OH, NH3+, 7O, 4OCH(3) groups in binding to DNA. Time-resolved fluorescence measurement and diffusion ordered spectroscopic studies reveal the formation of complex. The nonspecific interactions as well as those essential for biological activity are discussed along with its medicinal importance. PMID- 19285416 TI - Spatial distribution of (241)Am, (137)Cs, (238)Pu, (239,240)Pu and (241)Pu over 17 year periods in the Ravenglass saltmarsh, Cumbria, UK. AB - Ninety five surface scrape samples were collected at the Ravenglass saltmarsh and analysed for radionuclides by alpha spectrometry ((238)Pu and (239,240)Pu), gamma spectrometry ((241)Am and (137)Cs) and liquid scintillation counting ((241)Pu). Both (241)Am and (137)Cs activities are compared with those reported by Horrill [1983. Concentrations and spatial distribution of radioactivity in an ungrazed saltmarsh. In: Coughtrey, P.J. (Ed.), Ecological Aspects of Radionuclide Release. British Ecological Society Special Publication No. 3. Blackwell, Oxford, pp. 119 215.] Significant decreases in activities for both radionuclides were observed which is caused by the declining levels of discharges from the Sellafield nuclear reprocessing plant since the 1980s. It has been concluded that the spatial distribution of these radionuclides are controlled by the tidal currents and the clay contents in the sediments. There is evidence of surface erosion of the saltmarsh and redistribution of radionuclides in the saltmarsh using isotopic ratios of measured Pu. PMID- 19285417 TI - Calculation of isotope-specific exemption levels for surface contamination. AB - Isotope-specific exemption levels for surface contamination are calculated for representative radionuclides in general nuclear power plants by developing a deterministic dose assessment model for surface contamination that can be applied to radiation, transport and waste safety, and a practical idea of judging exemption for gross surface contamination by measuring gross gamma-ray emission has been proposed. In the dose assessment model, the objects with surface contamination are classified into three types: manually handled, closely handled and remotely handled objects, and the exemption criteria are chosen to be 0.01mSv/yr in the case of using realistic exposure parameters and 1mSv/yr in the case of using low-probability exposure parameters in accordance with the IAEA Safety Standards Series No. RS-G-1.7. Taking into account the distribution area of surface contamination assumed in the dose assessment model, instead of using the evaluation area of 100cm(2) without variation, the exemption levels for gross surface contamination are found to be higher than those obtained by the conventional method for some radionuclides such as Mn-54, Co-60, Zn-65, Nb-94, Cs 134, Cs-137, Eu-152 and Eu-154. PMID- 19285418 TI - Economic scale of utilization of radiation in medicine in Japan. AB - Economic scale of radioisotopes (RI) in Japan is studied in the field of medicine, agriculture and a part of industry. (1) RI is used during medical examination with economic scale by 1.7M$ (million dollars) in 1997 and 0.4M$ in 2005. (2) Economic scale of RI utilization in agriculture is 4M$ for R&D, 127M$ for environmental protection and 1M$ for chronology. RI usage in agriculture is increased five times due to needs at environmental technology lasted after the Kyoto protocol. (3) Indirect economic scale of RI ((85)Kr, (147)Pm, (90)Cr) usage in paper fabrication field in Japan for 2006 is 8432M$. PMID- 19285419 TI - Radioisotopic neutron transmission spectrometry: Quantitative analysis by using partial least-squares method. AB - Neutron spectrometry, based on the scattering of high energy fast neutrons from a radioisotope and slowing-down by the light hydrogen atoms, is a useful technique for non-destructive, quantitative measurement of hydrogen content because it has a large measuring volume, and is not affected by temperature, pressure, pH value and color. The most common choice for radioisotope neutron source is (252)Cf or (241)Am-Be. In this study, (252)Cf with a neutron flux of 6.3x10(6)n/s has been used as an attractive neutron source because of its high flux neutron and weak radioactivity. Pulse-height neutron spectra have been obtained by using in-house built radioisotopic neutron spectrometric system equipped with (3)He detector and multi-channel analyzer, including a neutron shield. As a preliminary study, polyethylene block (density of approximately 0.947g/cc and area of 40cmx25cm) was used for the determination of hydrogen content by using multivariate calibration models, depending on the thickness of the block. Compared with the results obtained from a simple linear calibration model, partial least-squares regression (PLSR) method offered a better performance in a quantitative data analysis. It also revealed that the PLSR method in a neutron spectrometric system can be promising in the real-time, online monitoring of the powder process to determine the content of any type of molecules containing hydrogen nuclei. PMID- 19285420 TI - (86)Y production via (86)Sr(p,n) for PET imaging at a cyclotron. AB - Excitation functions of (86)Y production via (86)Sr(p,xn), (86)Sr(d,xn), (85)Rb(alpha,xn), (85)Rb((3)He,xn), and (nat)Zr(d,alphaxn) reactions were studied by means of ALICE-ASH code and the results were compared with ALICE-91 code and experimental data. The greatest nuclear reaction of cyclotron (86)Y production was found out as (86)Sr(p,n)(86)Y process. (86)Y production yield was calculated too. A SrCO(3) thick film was deposited on a copper substrate by sedimentation method. The deposited (nat)SrCO(3) was irradiated with 15MeV proton at 30microA current beam. The separation of Y from Cu and Sr was carried out by means of dual ion exchange chromatography. PMID- 19285421 TI - Measurement of natural radionuclides in phosphgypsum using an anti-cosmic gamma ray spectrometer. AB - Gamma-ray spectrometry measurements have been carried out to determine the activity of natural radionuclides in a phosphogypsum sample included in a specific tight container. The gamma spectrometer includes an N-type coaxial high purity germanium (HPGe) detector equipped with an anti-cosmic system. This measurement required the determination of linear attenuation coefficients of phosphogypsum to calculate self-absorption correction between efficiency calibration conditions and measurement ones. The results are given for the three natural chains and for (40)K, in term of specific activity/g of dry material, ranging from a few Bq kg(-1) to a few hundreds Bq kg(-1). The equilibrium within the different families and the (235)U/(238)U ratio are discussed. PMID- 19285422 TI - Safety evaluation for regulatory management of Australian radioisotope production facilities. AB - This paper describes the approach of Australian Radiation Protection and Nuclear Safety Agency (ARPANSA) in the regulatory management of Radioisotope Production facilities in Australia. ARPANSA is the regulatory authority for commonwealth entities operating nuclear installations including Radioisotope Production facilities. In assessing the application for operating nuclear installations the ARPANSA assessors prepare a Safety Evaluation Report, which is a recommendation to the Chief Executive Officer (CEO) of ARPANSA whether to issue a licence to site, construct, operate and decommission facilities. In particular, the CEO must take into account international best practice in radiation protection and nuclear safety when making licence decisions. PMID- 19285423 TI - Coordination and speciation of cadmium in corn seedlings and its effects on macro and micronutrients uptake. AB - The effect of cadmium (Cd) on both the absorption of important nutrients and the synthesis of low molecular weight thiols (LMWTs) was investigated in corn plants. The inductively coupled plasma-optical emission spectroscopy results demonstrated that the concentration of Cd in tissues (mainly in roots) increased as the concentration in the medium increased. In addition, the concentration of phosphorus increased in roots of Cd treated plants but remained at normal concentration in shoots. On the other hand, the uptake of sulfur (S) followed a similar trend as the Cd uptake. The concentration of S and the production of LMWT were found to increase significantly upon exposure to Cd. The results of the X ray absorption spectroscopy analyses indicated that Cd within tissues was bound to S ligands with interatomic distances of 2.51-2.52 A. These results confirm a strong linkage between S uptake and the production of LMWT upon exposure to Cd. PMID- 19285424 TI - Nitric oxide treatment reduces neo-intimal formation and modulates osteopontin expression in an ex-vivo human model of intimal hyperplasia. AB - In this study the effects of nitric oxide (NO) on intimal hyperplasia (IH) were evaluated in an ex-vivo model of human saphenous vein (SV). SV segments were cultured in conditions able to reproduce IH (FCS), or in medium alone (RPMI), or in presence of a NO donor (NO). Osteopontin (OPN) and Interleukin (IL)-6 were determined in the medium at different culture times and in the tissue, at the end of experiment. OPN and IL-6 release in medium was increased in FCS with respect to RPMI (OPN: 13.9+/-2.9 vs. 2.3+/-0.8 microg/ml, p=0.0011; IL-6: 304.2+/-64.7 vs. 42.0+/-10.1 ng/ml, p<0.0006) as well as intima thickness, that positively correlated with OPN production (r=0.81). In tissue OPN was higher in FCS (82.0+/ 30.3 ng/mg protein) than in RPMI (13.8+/-4.2, p=0.0051) and at baseline (3.7+/ 0.7, p=0.018). NO reduces IH progression (25%) and both OPN and IL-6 expression (OPN/GAPDH: undetectable baseline; 0.27+/-0.06 RPMI; 0.89+/-0.28 FCS; 0.09+/-0.05 NO; p=0.026 FCS vs. baseline, p=0.018 vs. RPMI, p=0.005 vs. NO). The beneficial NO effect on IH reduction appears to be mediated by the indirect inhibition of OPN production. NO could modulates the initial inflammatory signals that induces the OPN over-production with the related cascade of events leading to IH. PMID- 19285425 TI - A novel vaccine adjuvant for recombinant flu antigens. AB - Many new vaccines under development consist of rationally designed recombinant proteins that are relatively poor immunogens unless combined with potent adjuvants. There is only one adjuvant in common use in the U.S., aluminum phosphate or hydroxide (e.g. alum). This adjuvant, however, has significant limitations, particularly regarding the generation of strong cell-mediated (T cell) immune responses. A novel adjuvant, JVRS-100, composed of cationic liposome DNA complexes (CLDC) has been evaluated for immune enhancing activity. The JVRS 100 adjuvant has been shown to elicit robust immune responses compared to CpG oligonucleotides, alum, and MPL adjuvants, and efficiently enhances both humoral and cellular immune responses. Safety has been evaluated in preclinical studies, and the adjuvant is now in early-stage clinical development. One application of this novel adjuvant is to augment the immune responses to recombinant subunit antigens, which are often poorly immunogenic. The JVRS-100 adjuvant, when combined with a recombinant influenza hemagglutinin (H1), elicited increased specific antibody and T-cell responses in mice. Single-dose vaccination and prime/boost vaccinations with JVRS-100-H1 were both shown to be protective (i.e., survival, reduced weight loss) following H1N1 (PR/8/34) virus challenge. Enhanced immunological responses could be critically important for improved efficacy and dose-sparing of a recombinant influenza vaccine. PMID- 19285426 TI - Single-dilution assay of new vaccines produced using plant and insect cell substrates. AB - Large numbers of animals are required in multi-dilution assays of vaccines produced in mammalian, plant and insect cell substrates. Animal vaccination and serum sampling require skilled labour, adding to testing costs. More tests are required with homologous reference preparations. Use of a single antigen reference to assay combined formulations is invalid due to synergistic or other effects of multiple antigens. Licensing authorities in Europe recommend use of single-dilution assays for some vaccines, together with homologous reference preparations. Vaccine and reference formulations in a single-dilution assay must have similar compositions to ensure similar dose response relationships. Design and validation of such a single-dilution assay is the main consideration of this article. Multi-dilution in vivo vaccine assays have inherent problems also affecting single-dilution assays. Statistical analysis of data from multi dilution assays requires significance tests of assay validity, but with a single dilution assay, some aspects of validity are assumed. These aspects include linearity and parallelism of the dilution-response curves for both reference and test preparations. This article shows that despite inherent problems associated with multi-dilution animal assays, the need to move to a simpler single-dilution model necessitates establishing minimal inherent conditions in the multi-dilution assay for a valid interpretation of the single-dilution assay. PMID- 19285427 TI - Vaccine production in Neurospora crassa. AB - We have chosen to use the filamentous fungus Neurospora crassa to produce subunit vaccines. Here we describe the production and purification of Influenza hemagglutinin and neuraminidase antigens in N. crassa. The N. crassa system used by Neugenesis offers many advantages over other systems for production of recombinant protein. In contrast to mammalian cell culture, N. crassa can be grown in a rapid and economic manner, generating large amounts of recombinant protein in simple, defined medium. Vaccines, therefore, can be produced more rapidly and at lower cost than conventional cell culture or egg-based systems. This has important applications to tailoring the seasonal vaccine supply and responding to new pandemics. PMID- 19285428 TI - Transesophageal echocardiography in detecting tricuspid valve pathology in an intensive care unit patient. PMID- 19285429 TI - A novel technique for easy identification of the subclavian vein during ultrasound-guided cannulation. PMID- 19285430 TI - An 11-year, single-institution analysis of intra-aortic balloon pump use in cardiac surgery. AB - OBJECTIVES: The primary objective of this study was to analyze perioperative intra-aortic balloon pump (IABP) insertion in patients undergoing cardiac surgery in the authors' institution from 1995 to 2005 and to propose an explanation for changes in use over this period. A secondary objective was to assess patient variables associated with IABP use. DESIGN: This is a retrospective study including patients who underwent cardiac surgery between 1995 and 2005. SETTING: The Cardiothoracic Anesthesia Patient Registry of a single teaching institution was queried to obtain the required information. PARTICIPANTS: Thirty thousand two hundred sixty-nine cardiac surgery patients. INTERVENTIONS: Intra-aortic balloon pump insertion before surgery, after cardiopulmonary bypass, or in the cardiovascular intensive care unit was assessed in patients who underwent isolated coronary artery bypass graft surgery, valve surgery, or both. Select patient variables were analyzed for their association with IABP insertion. Transesophageal echocardiography (TEE) examinations, milrinone use, and mortality rates also were determined. MEASUREMENTS AND MAIN RESULTS: Among 30,269 cardiac surgery patients, 1,310 (4.32%) underwent IABP insertion. Combined preoperative, intraoperative, and postoperative IABP use decreased from 7.8% in 1995 to 3.0% in 2005. Simultaneously, the intraoperative use of milrinone increased from 4.8% to 8.8% and postoperative use increased from 5.2% to 7.8%. The number of intraoperative TEE examinations more than doubled from approximately 1,700 to 3,500. The overall mortality for patients with preoperative, intraoperative, and postoperative IABP insertion was 12.6%, 17.5%, and 47.7%, respectively. CONCLUSIONS: From 1995 to 2005, preoperative, intraoperative, and postoperative IABP use decreased by approximately 60% in cardiac surgery patients. Simultaneously, the use of TEE and milrinone each doubled. Although a cause effect relationship cannot be established from the present study's observational data, the trends coincide and may be related. PMID- 19285431 TI - Acute cardiovascular collapse in total anomalous pulmonary venous connection. PMID- 19285432 TI - A cavity in the left ventricular outflow tract: a disastrous consequence of tooth decay? PMID- 19285433 TI - The impact of lung recruitment on hemodynamics during one-lung ventilation. AB - OBJECTIVE: The objective of the present study was to investigate respiratory and hemodynamic changes by measuring continuous cardiac output, cardiac filling, and stroke volume variation after lung recruitment in thoracic surgery. DESIGN: A prospective, observational study. SETTING: A tertiary care university hospital single institution. PARTICIPANTS: Forty patients undergoing thoracotomy for lung resection with at least 1 hour of one-lung ventilation (OLV). INTERVENTIONS: During OLV, an alveolar recruitment maneuver (ARM) was performed. MEASUREMENTS AND MAIN RESULTS: Based on Edwards Vigileo/FloTrac system (Edwards Lifesciences, Irvine, CA), the arterial pressure-based cardiac output, cardiac index, systemic vascular resistance, stroke volume variation, and central venous oxygen saturation were recorded immediately before the maneuver and 1, 2, 3, 4, 5, and 10 minutes after the maneuver. Stroke volume variation was the parameter most affected during and after the maneuver; it increased to 50% and 40% in the first and second minute, respectively (p < 0.01). The cardiac index was also affected and decreased 9.4% (p < 0.05) in the first minute after the maneuver. ScvO2 decreased significantly during the first 2 minutes (7% and 6.5%, respectively). However, after 3 minutes, all values recorded were similar to prerecruitment values. The PaO2 and PcvO2 from samples taken 10 minutes after the maneuver improved considerably with respect to the values before alveolar recruitment. CONCLUSIONS: The authors concluded that during open-chest surgery with OLV, an ARM effectively improved oxygenation without inducing important circulatory changes. PMID- 19285434 TI - Nonfatal intracardiac thromboembolism during liver transplantation. PMID- 19285435 TI - Sepsis and gas-forming splenic abscess by Clostridium septicum in a patient with type 2 diabetes. AB - Clostridium infections are rare but frequently associated with malignancy, and mortality approaches 100% if care is not rendered within 12 to 24 h. These infections are associated with various medical problems including diabetes mellitus. In this report, we describe a unique case of sepsis and a gas-forming splenic abscess caused by Clostridium septicum in a type 2 diabetes patient which was treatable solely with antibiotics. PMID- 19285436 TI - Cyclic adenosine monophosphate suppresses the transcription of proinflammatory cytokines via the phosphorylated c-Fos protein. AB - Intracellular cyclic adenosine monophosphate (cAMP) suppresses innate immunity by inhibiting proinflammatory cytokine production from monocytic cells. Enhanced expression of interleukin-10 (IL-10) has been suggested to be the mechanism of suppression. However, cAMP is still capable of suppressing production of the cytokines TNF-alpha and IL-12 in IL-10-deficient dendritic cells (DCs). Here, we demonstrated that the transcription factor c-Fos was responsible for the cAMP mediated suppression of inflammatory cytokine production. c-Fos accumulated at high amounts in response to cAMP and lipopolysaccharide (LPS). Overexpression of c-Fos suppressed LPS-induced cytokine production, whereas cAMP-mediated suppression of TNF-alpha and IL-12 was impaired in Fos(-/-) DCs or in RAW264.7 cells treated with c-Fos siRNA. c-Fos physically interacted with p65 protein and reduced the recruitment of p65 to the Tnf promoter. Multiple sites of c-Fos were phosphorylated by the IKKbeta protein. Thus, we propose that c-Fos is a substrate of IKKbeta and is responsible for the immunosuppressive effect of cAMP. PMID- 19285437 TI - Hepatic interleukin-7 expression regulates T cell responses. AB - Systemic cytokine activity in response to Toll-like receptor (TLR) signaling induces the expression of various proteins in the liver after infections. Here we show that Interleukin-7 (IL-7), the production of which was thought to occur at a constant rate in vivo, was a hepatically expressed protein that directly controled T cell responses. Depletion of IL-7 expression in the liver abrogated several TLR-mediated T cell events, including enhanced CD4+ T cell and CD8+ T cell survival, augmented CD8+ T cell cytotoxic activity, and the development of experimental autoimmune encephalitis, a Th17 cell-mediated autoimmune disease. Thus, T cell responses are regulated by hepatocyte-derived IL-7, which is expressed in response to TLR signaling in vivo. We suggested that TLR-induced IL 7 expression in the liver, which is an acute-phase response, may be a good diagnostic and therapeutic target for efficient vaccine developments and for conditions characterized by TLR-mediated T cell dysregulation, including autoimmune diseases. PMID- 19285438 TI - An essential role of the Forkhead-box transcription factor Foxo1 in control of T cell homeostasis and tolerance. AB - Members of the Forkhead box O (Foxo) family of transcription factors are key regulators of cellular responses, but their function in the immune system remains incompletely understood. Here we showed that T cell-specific deletion of Foxo1 gene in mice led to spontaneous T cell activation, effector T cell differentiation, autoantibody production, and the induction of inflammatory bowel disease in a transfer model. In addition, Foxo1 was critical for the maintenance of naive T cells in the peripheral lymphoid organs. Transcriptome analyses of T cells identified Foxo1-regulated genes encoding, among others, cell-surface molecules, signaling proteins, and nuclear factors that control gene expression. Functional studies validated interleukin-7 receptor-alpha as a Foxo1 target gene essential for Foxo1 maintenance of naive T cells. These findings reveal crucial functions of Foxo1-dependent transcription in control of T cell homeostasis and tolerance. PMID- 19285439 TI - The ubiquitin ligase RNF5 regulates antiviral responses by mediating degradation of the adaptor protein MITA. AB - Viral infection activates transcription factors NF-kappaB and IRF3, which collaborate to induce type I interferons (IFNs) and elicit innate antiviral response. MITA (also known as STING) has recently been identified as an adaptor that links virus-sensing receptors to IRF3 activation. Here, we showed that the E3 ubiquitin ligase RNF5 interacted with MITA in a viral-infection-dependent manner. Overexpression of RNF5 inhibited virus-triggered IRF3 activation, IFNB1 expression, and cellular antiviral response, whereas knockdown of RNF5 had opposite effects. RNF5 targeted MITA at Lys150 for ubiquitination and degradation after viral infection. Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER) and viral infection caused their redistribution to the ER and mitochondria, respectively. We further found that virus-induced ubiquitination and degradation of MITA by RNF5 occurred at the mitochondria. These findings suggest that RNF5 negatively regulates virus-triggered signaling by targeting MITA for ubiquitination and degradation at the mitochondria. PMID- 19285440 TI - Identification of two new Dichelobacter nodosus strains in Germany. AB - The multi-strain anaerobe Dichelobacter nodosus causes footrot, a contagious disease of the digits of ruminants that occurs worldwide. Eleven D. nodosus-like isolates from footrot lesions of sheep in Germany were investigated by PCR amplification of the variable region of the D. nodosus fimbrial (fimA) gene. In six isolates, fimA PCR amplicons of a size suggestive of D. nodosus were generated. Single-strand conformational polymorphism (SSCP) analysis of the amplicons revealed three banding patterns, the cloning and sequencing of which confirmed they represented three different strains of D. nodosus. One strain had a fimA sequence identical to D. nodosus isolate 265 of serotype H1, whilst the other two strains (B(de1) and H(de1)) exhibited novel fimA sequences. The B(de1) strain was most similar to D. nodosus serotype B1 isolate AC127, with six nucleotide differences in the fimA gene and four amino acid substitutions at the protein level. However, strain H(de1) was quite different from known strains of D. nodosus having 15 nucleotide differences and a three nucleotide deletion in its fimA gene relative to its closest homologue, isolate 351 of serotype H2. This resulted in 12 amino acid differences and one amino acid deletion at the protein level which suggests that H(de1) represents a new D. nodosus serotype within serogroup H. The identification of novel strains may have implications for current vaccination strategies against the disease. PMID- 19285441 TI - Unlocking the genetic make-up of canine hip dysplasia: we can work it out. PMID- 19285442 TI - Chylomicron retention disease: a long term study of two cohorts. AB - Lipoprotein assembly is critical for the intestinal absorption of dietary lipids and of fat-soluble vitamins. Through their inhibition of chylomicron secretion, mutations of the Sar1B gene coding for Sar1 GTPase are associated with chylomicron retention disease (CRD). The aim of this study was to describe the phenotypic expression of CRD in two clinically and genetically well characterized cohorts, and to compare their long term evolution. The study in 7 children from France (X age 11.3+/-1.7 years) and 9 from Quebec, Canada (X age 12+/-2.5 years) involved data collection from medical records for growth evaluation, neurological and ophthalmological status as well as bone density over an average follow-up period of 4.9 years for the French cohort and of 10.6 years for the Canadian one. All CRD patients presented within the first few months of life with diarrhea and failure to thrive. Severe hypocholesterolemia coupled with normal triglycerides was associated with low LDL and HDL-cholesterol, as well as with low apolipoproteins A-I and B. Varying degrees of essential fatty acid and of vitamin E deficiency were observed. The earlier diagnosis in the Canadian cohort (1.3+/ 0.04 years) than in the French one (6.3+/-1.3 years) was unrelated with the severity of presenting symptoms. The fact that the disease had more impact on growth and bone density in the latter group may be related to delayed diagnosis of the disease. Vitamin E deficiency led to functional neurological and ophthalmic changes in a small number of patients but only one developed areflexia. Finally, genotype-phenotype correlation is not obvious in our cohort with CRD; even if, the Canadian subjects with the allele 409G>A had a more severe degree (P<0.001) of hypocholesterolemia than the other patients, many clinical data are inconsistent with a hypothetical genotype-phenotype correlation. This study provides new insights on the phenotypic expression of CRD over time and emphasizes the need to screen the lipid profile of infants with chronic diarrhea and failure to thrive. PMID- 19285443 TI - Public trust and research a decade later: what have we learned since Jesse Gelsinger's death? AB - Almost a decade has passed since the untimely death of Jesse Gelsinger. The reflections of Dr. Wilson and efforts made on a national scale to address various ethical issues in biomedical research provide an opportunity to consider what progress has been made in efforts to build and restore the public's trust in biomedical research. The restoration of public trust is especially critical in the aftermath of tragic events like Mr. Gelsinger's death and the authors note the need for greater emphasis on building public trust than has occurred to date in the broader biomedical research community. PMID- 19285444 TI - Structural Insights into riboswitch control of the biosynthesis of queuosine, a modified nucleotide found in the anticodon of tRNA. AB - The modified nucleotide queuosine (Q) is almost universally found in the anticodon wobble position of specific tRNAs. In many bacteria, biosynthesis of Q is modulated by a class of regulatory mRNA elements called riboswitches. The preQ(1) riboswitch, found in the 5'UTR of bacterial genes involved in synthesis of the Q precursors preQ(0) and preQ(1), contains the smallest known aptamer domain. We report the solution structure of the preQ(1) riboswitch aptamer domain from Bacillus subtilis bound to preQ(1), which is a unique compact pseudoknot with three loops and two stems that encapsulates preQ(1) at the junction between the two stems. The pseudoknot only forms in the presence of preQ(1), and the 3' A rich tail of the aptamer domain is an integral part of the pseudoknot. In the absence of preQ(1), the A-rich tail forms part of the antiterminator. These structural studies provide insight into riboswitch transcriptional control of preQ(1) biosynthesis. PMID- 19285445 TI - A conserved WD40 protein binds the Cajal body localization signal of scaRNP particles. AB - Small Cajal body (CB)-specific RNPs (scaRNPs) function in posttranscriptional modification of small nuclear (sn)RNAs. An RNA element, the CAB box, facilitates CB localization of H/ACA scaRNPs. Using a related element in Drosophila C/D scaRNAs, we purified a fly WD40 repeat protein that UV crosslinks to RNA in a C/D CAB box-dependent manner and associates with C/D and mixed domain C/D-H/ACA scaRNAs. Its human homolog, WDR79, associates with C/D, H/ACA, and mixed domain scaRNAs, as well as with telomerase RNA. WDR79's binding to human H/ACA and mixed domain scaRNAs is CAB box dependent, and its association with mixed domain RNAs also requires the ACA motif, arguing for additional interactions of WDR79 with H/ACA core proteins. We demonstrate a requirement for WDR79 binding in the CB localization of a scaRNA. This and other recent reports establish WDR79 as a central player in the localization and processing of nuclear RNPs. PMID- 19285447 TI - [Needs of the family caregivers of stroke survivors in the home: structured review of the literature from 2000 to 2007]. AB - OBJECTIVE: To perform a structured review of the literature from 2000 to 2007 on the needs of the caregivers of stroke survivors in the postacute phase of the illness process at home. METHODS: Searches were conducted in the CINAHL, MEDLINE, EMBASE, PSYCHINFO, Cochrane Library Plus, CDSR (coch), DARE, CCTR, ACP Journal Club (ACP), IBECS, LILACS and IME databases using the terms "stroke", "caregiver" and "needs (assessment)". RESULTS: We selected 270 abstracts for review. Of these, only 53 met the inclusion criteria and just 12 achieved preestablished quality standards. Despite wide variability among the selected studies, the literature reviewed revealed that the two most prevalent needs for the caregivers of stroke survivors were information and support in the development of caregiving skills. Care for the caregiver herself, as well as the development and provision of support services, were defined as the two main areas where these participants seem to need support while adapting to and performing this newly adopted role. CONCLUSIONS: The studies reviewed show an increasing demand for support and care for stroke survivors' caregivers. Because of the highly diverse contexts of these studies, the lack of an explicit definition on the concept of "need", and the wide heterogeneity in caregivers' situations, summarizing the results of these studies is difficult. New studies are required in our context that take these limitations into account and try to overcome them. PMID- 19285448 TI - [Secularism at hospital]. PMID- 19285449 TI - Obesogenic environments: Are neighbourhood environments that limit physical activity obesogenic? AB - The impact of obesogenic environments on adolescent health is poorly understood. This study examines if neighbourhood features related to physical activity are also related to unhealthy weight status. Adolescents (N=4587, age 15-17 years, 51.4% male) self-reported physical activity and neighbourhood perceptions. Trained researchers measured height and weight. Logistic regression identified if neighbourhood perceptions predicted overweight or obesity. Adolescents who reported convenient physical activity facilities were 2% less likely to be overweight/obese and 5% less likely to be obese, controlled for socio-demographic variables and clustering by school. Physical activity does not appear to directly influence or mediate the relationship between perceived convenient facilities and weight status. PMID- 19285450 TI - Developmental aspects of sleep hygiene: findings from the 2004 National Sleep Foundation Sleep in America Poll. AB - OBJECTIVE: To examine the associations between sleep hygiene and sleep patterns in children ages newborn to 10 years. The relationships between key features of good sleep hygiene in childhood and recognizable outcomes have not been studied in large, nationally representative samples. PARTICIPANTS AND METHODS: A national poll of 1473 parents/caregivers of children ages newborn to 10 years was conducted in 2004. The poll included questions on sleep hygiene (poor sleep hygiene operationally defined as not having a consistent bedtime routine, bedtime after 9:00 PM, having a parent present when falling asleep at bedtime, having a television in the bedroom, and consuming caffeinated beverages daily) and sleep patterns (sleep onset latency, frequency of night wakings, and total sleep time). RESULTS: Across all ages, a late bedtime and having a parent present when the child falls asleep had the strongest negative association with reported sleep patterns. A late bedtime was associated with longer sleep onset latency and shorter total sleep time, whereas parental presence was associated with more night wakings. Those children (ages 3+) without a consistent bedtime routine also were reported to obtain less sleep. Furthermore, a television in the bedroom (ages 3+) and regular caffeine consumption (ages 5+) were associated with shorter total sleep time. CONCLUSIONS: Overall, this study found that good sleep hygiene practices are associated with better sleep across several age ranges. These findings support the importance of common US based recommendations that children of all ages should fall asleep independently, go to bed before 9:00 PM, have an established bedtime routine, include reading as part of their bedtime routine, refrain from caffeine, and sleep in bedrooms without televisions. PMID- 19285451 TI - Treatment using oral appliance for obstructive sleep apnea syndrome patients with multiple missing teeth. PMID- 19285452 TI - The delirium observation screening scale recognizes delirium early after cardiac surgery. AB - BACKGROUND: Delirium or acute confusion is a temporary mental disorder which occurs frequently among hospitalized elderly patients. Patients who undergo cardiac surgery have an increased risk of developing delirium. Prevention or early recognition of delirium is essential. The Delirium Observation Screening (DOS) scale was developed to facilitate early recognition of delirium by nurses' observations during routine clinical care. AIM: The aim of this study was to validate the DOS scale in accordance with the diagnosis of the psychiatrist, using the DSM-IV criteria as the gold standard. METHODS: In this observational study, the DOS scale was used to assess whether 112 patients who underwent elective cardiac surgery had developed a postoperative delirium. The psychiatrist was consulted to confirm or refute the diagnosis delirium. Wilcoxon's Rank Sum Test was utilized to compare patients with and without delirium on duration of hospital stay. A Receiver Operating Characteristic Curve of the DOS scale was constructed with accompanying Area Under the Curve (AUC). RESULTS: Based on the diagnosis of the psychiatrist, the incidence of delirium following cardiac surgery was 21.4% and the mean duration of delirium was two and a half days. The time to discharge was 11 days longer in patients with delirium. In 27 of the 112 patients a DOS score of >or=3 was found, that indicates delirium. The sensitivity and specificity of the DOS scale was 100% and 96.6% respectively. The AUC was 0.98. CONCLUSION: The DOS scale is a very good instrument to facilitate early recognition of delirium by nurses' observation of patients who undergo cardiac surgery. Early recognition will expedite good postoperative management such as implementation of appropriate interventions, and may decrease negative consequences caused by postoperative delirium. PMID- 19285453 TI - Protection from radiation-induced pneumonitis using cerium oxide nanoparticles. AB - In an effort to combat the harmful effects of radiation exposure, we propose that rare-earth cerium oxide (CeO(2)) nanoparticles (free-radical scavengers) protect normal tissue from radiation-induced damage. Preliminary studies suggest that these nanoparticles may be a therapeutic regenerative nanomedicine that will scavenge reactive oxygen species, which are responsible for radiation-induced cell damage. The effectiveness of CeO(2) nanoparticles in radiation protection in murine models during high-dose radiation exposure is investigated, with the ultimate goal of offering a new approach to radiation protection, using nanotechnology. We show that CeO(2) nanoparticles are well tolerated by live animals, and they prevent the onset of radiation-induced pneumonitis when delivered to live animals exposed to high doses of radiation. In the end, these studies provide a tremendous potential for radioprotection and can lead to significant benefits for the preservation of human health and the quality of life for humans receiving radiation therapy. PMID- 19285454 TI - The dmNTS is not the source of increased blood pressure variability in baroreflex denervated rats. AB - A consistent and prominent feature, observed across many species, including our neuromuscular blocked (NMB) rat preparation, is that obliterating the baroafferent inputs to the brainstem, e.g., by sinoaortic denervation (SAD), significantly increases blood pressure variability (BPV). The sources of the BPV, however, are not completely understood, but involve both the central and the peripheral mechanisms. The key central noise source is likely in the brainstem. Previously, in NMB rats, we showed that the maximum gain of the baroreflex system is in the very low frequency (VLF) range of 0.01-0.2 Hz. In this study, using the same NMB preparation, we demonstrated that, after SAD, there was a significant increase in the VLF power of the expiratory systolic blood pressure (EsBP) spectrum, but a decrease in the VLF power of the expiratory heart inter-beat interval (EIBI) spectrum. Because dmNTS is the only major common anatomic node for the vascular sympathetic and the cardiac parasympathetic pathways, the opposite changes in the post-SAD VLF powers of the EsBP and EIBI spectra suggest that dmNTS is unlikely the major noise source for the post-SAD BPV. Supporting this finding, we found that the dmNTS evoked response to single pulse baroreflex afferent aortic depressor nerve (ADN) stimuli was substantially more reliable than the evoked systolic blood pressure responses to the same stimuli. PMID- 19285455 TI - Antibacterial activity of nanocomposites of silver and bacterial or vegetable cellulosic fibers. AB - Cellulose/Ag nanocomposites were prepared using two distinct methodologies and two cellulose substrates: vegetable and bacterial cellulose. These nanocomposites were characterized in terms of their morphology and chemical composition. Detailed studies on the antibacterial activity of these materials were carried out for Bacillus subtilis, Staphylococcus aureus and Klebsiella pneumoniae. Silver nanoparticles present in the cellulosic fibers in concentrations as low as 5.0x10(-4)wt.% make these nanocomposites effective antibacterial materials. We anticipate that the versatile use of these cellulose-based nanocomposites can bring a promising strategy to produce a wide range of interesting materials where antibacterial properties are crucial. PMID- 19285456 TI - Preparation and properties of macroporous brushite bone cements. AB - In the present work a macroporous brushite bone cement for use either as an injected or mouldable paste, or in the shape of preformed grafts, has been investigated. Macropores have been introduced by adding to the powder single crystals of mannitol which worked as a porogen. The size of the crystals was in the range of 250-500microm in diameter, suitable for cell infiltration, with a shape ratio between 3 and 6. From compression tests on cylindrical samples an elastic modulus in the range 2.5-4.2GPa and a compressive strength in the range 17.5-32.6MPa were obtained for a volume fraction of macropores varying between 15 and 0%. Thus the compressive strength exceeded in all tests the maximum value currently attributed to cancellous bone. PMID- 19285457 TI - Infection and stillbirth. AB - Infection may cause stillbirth by several mechanisms, including direct infection, placental damage, and severe maternal illness. Various organisms have been associated with stillbirth, including many bacteria, viruses, and protozoa. In developed countries, between 10% and 25% of stillbirths may be caused by an infection, whereas in developing countries, which have much higher stillbirth rates, the contribution of infection is much greater. In developed countries, ascending bacterial infection, both before and after membrane rupture, with organisms such as Escherichia coli, group B streptococci, and Ureaplasma urealyticum is usually the most common infectious cause of stillbirth. However, in areas where syphilis is prevalent, up to half of all stillbirths may be caused by this infection alone. Malaria may be an important cause of stillbirth in women infected for the first time in pregnancy. The two most important viral causes of stillbirth are parvovirus and Coxsackie virus, although a number of other viral infections appear to be causal. Toxoplasma gondii, Listeria monocytogenes, and the organisms that cause leptospirosis, Q fever, and Lyme disease have all been implicated as etiologic for stillbirth. In certain developing countries, the stillbirth rate is high and the infection-related component so great that achieving a substantial reduction in stillbirth should be possible by reducing maternal infections. However, because infection-related stillbirth is uncommon in developed countries, and because those that do occur are caused by a wide variety of organisms, reducing this etiologic component of stillbirth much further will be difficult. PMID- 19285458 TI - It was nice not seeing you: perceptual learning with rewards in the absence of awareness. AB - In this issue of Neuron, Seitz et al. show that humans exhibit enhanced perceptual discrimination for visual stimuli that have been repeatedly paired with reward under conditions of suppressed awareness. These findings challenge the view that awareness and focused attention are necessary for perceptual learning. PMID- 19285459 TI - Synaptogenic proteins and synaptic organizers: "many hands make light work". AB - Synaptogenesis is thought to be mediated by cell adhesion proteins, which induce the initial contact between an axon and its target cell and subsequently recruit and organize the presynaptic and postsynaptic protein machinery required for synaptic transmission. A new study by Linhoff and colleagues in this issue of Neuron identifies adhesion proteins of the LRRTM family as novel synaptic organizers. PMID- 19285460 TI - The headache of a hyperactive calcium channel. AB - Migraine is thought to be triggered by excessive neocortical neuronal excitability that leads to cortical spreading depression. In this issue of Neuron, Tottene et al. study a mouse model of familial hemiplegic migraine type 1, and provide evidence for the hyperactivity of P/Q-type calcium channel mediated cortical glutamatergic synaptic transmission as an underlying mechanism for the susceptibility of cortical spreading depression initiation in migraine disorders. PMID- 19285461 TI - Signaling aversive events in the midbrain: worse than expected. AB - Although we know a great deal about how the brain processes information about aversive and appetitive stimuli, it is not clear how these two systems interact to guide behavior. In this issue of Neuron, Jhou and colleagues identify a region in the midbrain tegmentum that signals aversive events and inhibits midbrain dopamine neurons. PMID- 19285462 TI - Perception and the medial temporal lobe: evaluating the current evidence. AB - A dominant view in the learning and memory literature states that a subset of anatomically related structures within the medial temporal lobe (MTL), including the hippocampus, entorhinal, perirhinal, and parahippocampal cortices, forms a functionally related system specialized for declarative memory but not for perception. However, recent reports challenge this view, suggesting instead that the medial temporal lobe is not only important for memory, but also critical for certain forms of perception. In this review, I argue that little or no conclusive evidence currently exists to support the latter view. Experimental studies that have examined the perceptual functions of the MTL in monkeys are inconclusive because they fail to isolate perceptual from mnemonic task demands. Evaluation of conflicting results from studies in human amnesic patients suggests that extraneous damage to extra-MTL areas may underlie the reported perceptual deficits in the group of amnesic patients at the heart of this debate. See the related Review from Baxter, "Involvement of Medial Temporal Lobe Structures in Memory and Perception," in this issue of Neuron. PMID- 19285463 TI - Involvement of medial temporal lobe structures in memory and perception. AB - Beginning approximately a decade and a half ago, it was suggested that some structures that are considered to be part of the "medial temporal lobe memory system" could play a role in perception as well. The implications of this view, interpreted broadly, are that medial temporal lobe structures may be understood as an extension of the ventral visual stream and that their functions cannot be described exclusively in terms of memory. Considerable evidence now supports the view that medial temporal lobe structures are involved in nonmnemonic aspects of cognition, such as perception. This discovery allows for a fuller understanding of the involvement of these structures in mental phenomena than does a purely mnemonic account of their function. See the related review by Suzuki, "Perception and the Medial Temporal Lobe: Evaluating the Current Evidence," in this issue of Neuron. PMID- 19285464 TI - Memory, perception, and the medial temporal lobe: a synthesis of opinions. PMID- 19285465 TI - Lesion mapping of cognitive abilities linked to intelligence. AB - The Wechsler Adult Intelligence Scale (WAIS) assesses a wide range of cognitive abilities and impairments. Factor analyses have documented four underlying indices that jointly comprise intelligence as assessed with the WAIS: verbal comprehension (VCI), perceptual organization (POI), working memory (WMI), and processing speed (PSI). We used nonparametric voxel-based lesion-symptom mapping in 241 patients with focal brain damage to investigate their neural underpinnings. Statistically significant lesion-deficit relationships were found in left inferior frontal cortex for VCI, in left frontal and parietal cortex for WMI, and in right parietal cortex for POI. There was no reliable single localization for PSI. Statistical power maps and cross-validation analyses quantified specificity and sensitivity of the index scores in predicting lesion locations. Our findings provide comprehensive lesion maps of intelligence factors, and make specific recommendations for interpretation and application of the WAIS to the study of intelligence in health and disease. PMID- 19285466 TI - Quantitative mapping of a digenic behavioral trait implicates globin variation in C. elegans sensory behaviors. AB - Most heritable behavioral traits have a complex genetic basis, but few multigenic traits are understood at a molecular level. Here we show that the C. elegans strains N2 and CB4856 have opposite behavioral responses to simultaneous changes in environmental O(2) and CO(2). We identify two quantitative trait loci (QTL) that affect this trait and map each QTL to a single-gene polymorphism. One gene, npr-1, encodes a previously described neuropeptide receptor whose high activity in N2 promotes CO(2) avoidance. The second gene, glb-5, encodes a neuronal globin domain protein whose high activity in CB4856 modifies behavioral responses to O(2) and combined O(2)/CO(2) stimuli. glb-5 acts in O(2)-sensing neurons to increase O(2)-evoked calcium signals, implicating globins in sensory signaling. An analysis of wild C. elegans strains indicates that the N2 alleles of npr-1 and glb-5 arose recently in the same strain background, possibly as an adaptation to laboratory conditions. PMID- 19285467 TI - Rewards evoke learning of unconsciously processed visual stimuli in adult humans. AB - The study of human learning is complicated by the myriad of processing elements involved in conducting any behavioral task. In the case of visual perceptual learning, there has been significant controversy regarding the task processes that guide the formation of this learning. However, there is a developing consensus that top-down, task-related factors are required for such learning to take place. Here we challenge this idea by use of a novel procedure in which human participants, who were deprived of food and water, passively viewed visual stimuli while receiving occasional drops of water as rewards. Visual orientation stimuli, which were temporally paired with the liquid rewards, were viewed monocularly and rendered imperceptible by continuously flashing contour-rich patterns to the other eye. Results show that visual learning can be formed in human adults through stimulus-reward pairing in the absence of a task and without awareness of the stimulus presentation or reward contingencies. PMID- 19285468 TI - Motor neurons with axial muscle projections specified by Wnt4/5 signaling. AB - Axial muscles are innervated by motor neurons of the median motor column (MMC). In contrast to the segmentally restricted motor columns that innervate limb, body wall, and neuronal targets, MMC neurons are generated along the entire length of the spinal cord. We show that the specification of MMC fate involves a dorsoventral signaling program mediated by three Wnt proteins (Wnt4, Wnt5a, and Wnt5b) expressed in and around the floor plate. These Wnts appear to establish a ventral(high) to dorsal(low) signaling gradient and promote MMC identity and connectivity by maintaining expression of the LIM homeodomain proteins Lhx3/4 in spinal motor neurons. Elevation of Wnt4/5 activity generates additional MMC neurons at the expense of other motor neuron columnar subtypes, whereas depletion of Wnt4/5 activity inhibits the production of MMC neurons. Thus, two dorsoventral signaling pathways, mediated by Shh and Wnt4/5, are required to establish an early binary divergence in motor neuron columnar identity. PMID- 19285469 TI - Wnt signals organize synaptic prepattern and axon guidance through the zebrafish unplugged/MuSK receptor. AB - Early during neuromuscular development, acetylcholine receptors (AChRs) accumulate at the center of muscle fibers, precisely where motor growth cones navigate and synapses eventually form. Here, we show that Wnt11r binds to the zebrafish unplugged/MuSK ectodomain to organize this central muscle zone. In the absence of such a zone, prepatterned AChRs fail to aggregate and, as visualized by live-cell imaging, growth cones stray from their central path. Using inducible unplugged/MuSK transgenes, we show that organization of the central muscle zone is dispensable for the formation of neural synapses, but essential for AChR prepattern and motor growth cone guidance. Finally, we show that blocking noncanonical dishevelled signaling in muscle fibers disrupts AChR prepatterning and growth cone guidance. We propose that Wnt ligands activate unplugged/MuSK signaling in muscle fibers to restrict growth cone guidance and AChR prepatterns to the muscle center through a mechanism reminiscent of the planar cell polarity pathway. PMID- 19285470 TI - An unbiased expression screen for synaptogenic proteins identifies the LRRTM protein family as synaptic organizers. AB - Delineating the molecular basis of synapse development is crucial for understanding brain function. Cocultures of neurons with transfected fibroblasts have demonstrated the synapse-promoting activity of candidate molecules. Here, we performed an unbiased expression screen for synaptogenic proteins in the coculture assay using custom-made cDNA libraries. Reisolation of NGL-3/LRRC4B and neuroligin-2 accounts for a minority of positive clones, indicating that current understanding of mammalian synaptogenic proteins is incomplete. We identify LRRTM1 as a transmembrane protein that induces presynaptic differentiation in contacting axons. All four LRRTM family members exhibit synaptogenic activity, LRRTMs localize to excitatory synapses, and artificially induced clustering of LRRTMs mediates postsynaptic differentiation. We generate LRRTM1(-/-) mice and reveal altered distribution of the vesicular glutamate transporter VGLUT1, confirming an in vivo synaptic function. These results suggest a prevalence of LRR domain proteins in trans-synaptic signaling and provide a cellular basis for the reported linkage of LRRTM1 to handedness and schizophrenia. PMID- 19285471 TI - Timescales of inference in visual adaptation. AB - Adaptation is a hallmark of sensory function. Adapting optimally requires matching the dynamics of adaptation to those of changes in the stimulus distribution. Here we show that the dynamics of adaptation in the responses of mouse retinal ganglion cells depend on stimulus history. We hypothesized that the accumulation of evidence for a change in the stimulus distribution controls the dynamics of adaptation, and developed a model for adaptation as an ongoing inference problem. Guided by predictions of this model, we found that the dynamics of adaptation depend on the discriminability of the change in stimulus distribution and that the retina exploits information contained in properties of the stimulus beyond the mean and variance to adapt more quickly when possible. PMID- 19285472 TI - Enhanced excitatory transmission at cortical synapses as the basis for facilitated spreading depression in Ca(v)2.1 knockin migraine mice. AB - Migraine is a common disabling brain disorder. A subtype of migraine with aura (familial hemiplegic migraine type 1: FHM1) is caused by mutations in Ca(V)2.1 (P/Q-type) Ca(2+) channels. Knockin mice carrying a FHM1 mutation show increased neuronal P/Q-type current and facilitation of induction and propagation of cortical spreading depression (CSD), the phenomenon that underlies migraine aura and may activate migraine headache mechanisms. We studied cortical neurotransmission in neuronal microcultures and brain slices of FHM1 mice. We show gain of function of excitatory neurotransmission due to increased action potential-evoked Ca(2+) influx and increased probability of glutamate release at pyramidal cell synapses but unaltered inhibitory neurotransmission at fast spiking interneuron synapses. Using an in vitro model of CSD, we show a causative link between enhanced glutamate release and CSD facilitation. The synapse specific effect of FHM1 mutations points to disruption of excitation-inhibition balance and neuronal hyperactivity as the basis for episodic vulnerability to CSD ignition in migraine. PMID- 19285473 TI - Differential effects of excitatory and inhibitory plasticity on synaptically driven neuronal input-output functions. AB - Ultimately, whether or not a neuron produces a spike determines its contribution to local computations. In response to brief stimuli the probability a neuron will fire can be described by its input-output function, which depends on the net balance and timing of excitatory and inhibitory currents. While excitatory and inhibitory synapses are plastic, most studies examine plasticity of subthreshold events. Thus, the effects of concerted regulation of excitatory and inhibitory synaptic strength on neuronal input-output functions are not well understood. Here, theoretical analyses reveal that excitatory synaptic strength controls the threshold of the neuronal input-output function, while inhibitory plasticity alters the threshold and gain. Experimentally, changes in the balance of excitation and inhibition in CA1 pyramidal neurons also altered their input output function as predicted by the model. These results support the existence of two functional modes of plasticity that can be used to optimize information processing: threshold and gain plasticity. PMID- 19285474 TI - The rostromedial tegmental nucleus (RMTg), a GABAergic afferent to midbrain dopamine neurons, encodes aversive stimuli and inhibits motor responses. AB - Separate studies have implicated the lateral habenula (LHb) or amygdala-related regions in processing aversive stimuli, but their relationships to each other and to appetitive motivational systems are poorly understood. We show that neurons in the recently identified GABAergic rostromedial tegmental nucleus (RMTg), which receive a major LHb input, project heavily to midbrain dopamine neurons, and show phasic activations and/or Fos induction after aversive stimuli (footshocks, shock predictive cues, food deprivation, or reward omission) and inhibitions after rewards or reward-predictive stimuli. RMTg lesions markedly reduce passive fear behaviors (freezing, open-arm avoidance) dependent on the extended amygdala, periaqueductal gray, or septum, all regions that project directly to the RMTg. In contrast, RMTg lesions spare or enhance active fear responses (treading, escape) in these same paradigms. These findings suggest that aversive inputs from widespread brain regions and stimulus modalities converge onto the RMTg, which opposes reward and motor-activating functions of midbrain dopamine neurons. PMID- 19285475 TI - Short-term memory for figure-ground organization in the visual cortex. AB - Whether the visual system uses a buffer to store image information and the duration of that storage have been debated intensely in recent psychophysical studies. The long phases of stable perception of reversible figures suggest a memory that persists for seconds. But persistence of similar duration has not been found in signals of the visual cortex. Here, we show that figure-ground signals in the visual cortex can persist for a second or more after the removal of the figure-ground cues. When new figure-ground information is presented, the signals adjust rapidly, but when a figure display is changed to an ambiguous edge display, the signals decay slowly--a behavior that is characteristic of memory devices. Figure-ground signals represent the layout of objects in a scene, and we propose that a short-term memory for object layout is important in providing continuity of perception in the rapid stream of images flooding our eyes. PMID- 19285476 TI - CotQuest: improved algorithm and software for nonlinear regression analysis of DNA reassociation kinetics data. AB - Cot analysis (DNA reassociation kinetics) has long been used to explore genome structure in individual species, estimate genome similarity among organisms, and evaluate diversity in ecological samples, yet the algorithms and computational tools designed for analyzing Cot data are outdated, difficult to use, and prone to error. We report a new nonlinear regression procedure for analysis of Cot data and describe our algorithms in detail. Our procedure is implemented as CotQuest, a suite of scripts designed for use with the statistics package SAS. Unlike previous programs, CotQuest does not require users to input guesses as to the final values of parameters; rather, it employs a novel algorithm to step through a sequence of progressively more complex models, with the results from a given analysis being used to generate starting values for the next model. Moreover, CotQuest returns a statistical comparison of potential models and provides a variety of model assessment and selection diagnostics to help users in model selection. In situations where two models possess similar goodness-of-fit assessments, visual analysis of the Cot curves and comparison of CotQuest generated graphs and statistics reflecting the normality and homoscedasticity of residuals can be employed to make educated choices between models. PMID- 19285477 TI - Application of retrovirus-mediated expression cloning for receptor screening of a parasite. AB - Retrovirus-mediated expression cloning has been applied in both virology and cell biology. Although there is some difficulty in applying this technique to screening for a receptor recognized by an intracellular parasite, we modified the conventional method to identify a putative receptor for the Plasmodium falciparum BAEBL protein. We show that this method is effective in screening for a parasite receptor. PMID- 19285478 TI - Liver type fatty acid binding protein (L-FABP) gene ablation reduces nuclear ligand distribution and peroxisome proliferator-activated receptor-alpha activity in cultured primary hepatocytes. AB - The effect of liver type fatty acid binding protein (L-FABP) gene ablation on the uptake and distribution of long chain fatty acids (LCFA) to the nucleus by real time laser scanning confocal imaging and peroxisome proliferator-activated receptor-alpha (PPARalpha) activity was examined in cultured primary hepatocytes from livers wild-type L-FABP+/+ and gene ablated L-FABP-/- mice. Cultured primary hepatocytes from livers of L-FABP-/- mice exhibited: (i) reduced oxidation of palmitic acid, a common dietary long chain fatty acid (LCFA); (ii) reduced expression of fatty acid oxidative enzymes-proteins transcriptionally regulated by PPARalpha; (iii) reduced palmitic acid-induced PPARalpha co immunoprecipitation with coactivator SRC-1 concomitant with increased PPARalpha co-immunoprecipitation with coinhibitor N-CoR; (iv) reduced palmitic acid-induced PPARalpha. Diminished PPARalpha activation in L-FABP null hepatocytes was associated with lower uptake of common dietary LCFA (palmitic acid as well as its fluorescent derivative BODIPY FL C(16)), reduced level of total unesterified LCFA, and real-time redistribution of BODIPY FL C(16) from the central nucleoplasm to the nuclear envelope. Taken together, these studies support the hypothesis that L-FABP may facilitate ligand (LCFA)-activated PPARalpha transcriptional activity at least in part by increasing total LCFA ligand available to PPARalpha for inducing PPARalpha-mediated transcription of proteins involved in LCFA metabolism. PMID- 19285479 TI - Role of mitochondria-associated hexokinase II in cancer cell death induced by 3 bromopyruvate. AB - It has long been observed that cancer cells rely more on glycolysis to generate ATP and actively use certain glycolytic metabolic intermediates for biosynthesis. Hexokinase II (HKII) is a key glycolytic enzyme that plays a role in the regulation of the mitochondria-initiated apoptotic cell death. As a potent inhibitor of hexokinase, 3-bromopyruvate (3-BrPA) is known to inhibit cancer cell energy metabolism and trigger cell death, supposedly through depletion of cellular ATP. The current study showed that 3-BrPA caused a covalent modification of HKII protein and directly triggered its dissociation from mitochondria, leading to a specific release of apoptosis-inducing factor (AIF) from the mitochondria to cytosol and eventual cell death. Co-immunoprecipitation revealed a physical interaction between HKII and AIF. Using a competitive peptide of HKII, we showed that the dissociation of hexokinase II from mitochondria alone could cause apoptotic cell death, especially in the mitochondria-deficient rho(0) cells that highly express HKII. Interestingly, the dissociation of HKII itself did not directly affect the mitochondrial membrane potential, ROS generation, and oxidative phosphorylation. Our study suggests that the physical association between HKII and AIF is important for the normal localization of AIF in the mitochondria, and disruption of this protein complex by 3-BrPA leads to their release from the mitochondria and eventual cell death. PMID- 19285480 TI - Contribution of organic anion transporting polypeptide OATP2B1 to amiodarone accumulation in lung epithelial cells. AB - The accumulation mechanisms of amiodarone (AMD) involving transporters in lung alveolar epithelial type II cells were studied. The uptake of AMD was examined using human alveolar epithelial-derived cell line A549 as a model. AMD was transported by the carrier-mediated system, and the apparent K(m) and V(max) values were 66.8+/-30.3 muM and 49.7+/-9.7 nmol/mg protein/5 min, respectively. The uptake of AMD by A549 cells was Na(+)-independent and was inhibited by substrates of human organic anion transporting polypeptide (OATP). The inhibition profiles were similar to the inhibitory effects of several compounds on OATP2B1 mediated E-3-S transport, and RT-PCR analysis showed mRNA expression of OATP2B1 and 1B3 in A549 cells. SiRNAs targeted to the OATP2B1 gene decreased the OATP2B1 mRNA expression level in A549 cells up to about 50% and reduced the uptake of AMD up to about 40%. These results indicate that AMD uptake mediated by carriers, including OATP2B1, might lead to accumulation of AMD in the lung and AMD-induced pulmonary toxicity (AIPT). PMID- 19285481 TI - Cell specificity, anti-inflammatory activity, and plausible bactericidal mechanism of designed Trp-rich model antimicrobial peptides. AB - To develop novel short Trp-rich antimicrobial peptides (AMPs) with potent cell specificity (targeting bacteria but not eukaryotic cells) and anti-inflammatory activity, a series of 11-meric Trp-rich model peptides with different ratios of Leu and Lys/Arg residues, XXWXXWXXWXX-NH(2) (X indicates Leu or Lys/Arg), was synthesized. K(6)L(2)W(3) displayed an approximately 40-fold increase in cell specificity, compared with the natural Trp-rich AMP indolicidin (IN). Lys containing peptides (K(8)W(3), K(7)LW(3) and K(6)L(2)W(3)) showed approximately 2 to 4-fold higher cell specificities than did their counterparts, the Arg containing peptides (R(8)W(3), R(7)LW(3) and R(6)L(2)W(3)), indicating that multiple Lys residues are more important than multiple Arg residues in the design of AMPs with good cell specificity. The excellent resistance of d-enantiomers (K(6)L(2)W(3)-D and R(6)L(2)W(3)-D) and Orn/Nle-containing peptides (O(6)L(2)W(3) and O(6)L(2)W(3)) to trypsin digestion compared with the rapid breakdown of the l enantiomers (K(6)L(2)W(3) and R(6)L(2)W(3)), highlights the clinical potential of such peptides. K(6)L(2)W(3), R(6)L(2)W(3), K(6)L(2)W(3)-D and R(6)L(2)W(3)-D caused weak dye leakage from bacterial membrane-mimicking negatively charged EYPG/EYPE (7:3, v/v) liposomes. Confocal microscopy showed that these peptides penetrated the cell membrane of Escherichia coli and accumulated in the cytoplasm, as observed for buforin-2. Gel retardation studies revealed that the peptides bound more strongly to DNA than did IN. These results suggested that one possible peptide bactericidal mechanism may relate to the inhibition of intracellular functions via interference with DNA/RNA synthesis. Furthermore, some model peptides, containing K(6)L(2)W(3), K(5)L(3)W(3), R(6)L(2)W(3), O(6)L(2)W(3), O(6)L(2)W(3), and K(6)L(2)W(3)-D inhibited LPS-induced inducible nitric oxide synthase (iNOS) mRNA expression, the release of nitric oxide (NO) following LPS stimulation in RAW264.7 cells and had powerful LPS binding activities at bactericidal concentrations. Collectively, our results indicated that these peptides have potential for future development as novel antimicrobial and anti-inflammatory agents. PMID- 19285482 TI - The yeast mitochondrial carrier proteins Mrs3p/Mrs4p mediate iron transport across the inner mitochondrial membrane. AB - The yeast proteins Mrs3p and Mrs4p are two closely related members of the mitochondrial carrier family (MCF), which had previously been implicated in mitochondrial Fe(2+) homeostasis. A vertebrate Mrs3/4 homologue named mitoferrin was shown to be essential for erythroid iron utilization and proposed to function as an essential mitochondrial iron importer. Indirect reporter assays in isolated yeast mitochondria indicated that the Mrs3/4 proteins are involved in mitochondrial Fe(2+) utilization or transport under iron-limiting conditions. To have a more direct test for Mrs3/4p mediated iron uptake into mitochondria we studied iron (II) transport across yeast inner mitochondrial membrane vesicles (SMPs) using the iron-sensitive fluorophore PhenGreen SK (PGSK). Wild-type SMPs showed rapid uptake of Fe(2+) which was driven by the external Fe(2+) concentration and stimulated by acidic pH. SMPs from the double deletion strain mrs3/4Delta failed to show this rapid Fe(2+) uptake, while SMPs from cells overproducing Mrs3/4p exhibited increased Fe(2+) uptake rates. Cu(2+) was transported at similar rates as Fe(2+), while other divalent cations, such as Zn(2+) and Cd(2+) apparently did not serve as substrates for the Mrs3/4p transporters. We conclude that the carrier proteins Mrs3p and Mrs4p transport Fe(2+) across the inner mitochondrial membrane. Their activity is dependent on the pH gradient and it is stimulated by iron shortage. PMID- 19285483 TI - Voltage-gated proton channel is expressed on phagosomes. AB - Voltage-gated proton channel has been suggested to help NADPH oxidase activity during respiratory burst of phagocytes through its activities of compensating charge imbalance and regulation of pH. In phagocytes, robust production of reactive oxygen species occurs in closed membrane compartments, which are called phagosomes. However, direct evidence for the presence of voltage-gated proton channels in phagosome has been lacking. In this study, the expression of voltage gated proton channels was studied by Western blot with the antibody specific to the voltage-sensor domain protein, VSOP/Hv1, that has recently been identified as the molecular correlate for the voltage-gated proton channel. Phagosomal membranes of neutrophils contain VSOP/Hv1 in accordance with subunits of NADPH oxidases, gp91, p22, p47 and p67. Superoxide anion production upon PMA activation was significantly reduced in neutrophils from VSOP/Hv1 knockout mice. These are consistent with the idea that voltage-gated proton channels help NADPH oxidase in phagocytes to produce reactive oxygen species. PMID- 19285484 TI - A South African family with the mitochondrial A1555G mutation on haplogroup L0d. AB - The most common mutation associated with aminoglycoside-induced deafness is A1555G and it has been found in diverse populations worldwide. In the present study we investigated a large South African family known to harbour A1555G. A total of 97 family members were genotyped using the SNaPshot technique and 76 were found to be A1555G-positive (on haplogroup L0d) and are therefore at risk of developing irreversible hearing loss. The method worked equally well on both blood (from adults) and buccal swabs (from children). Variants in the tRNA(Ser(UCN)), A10S in TRMU and 35delG in GJB2 genes were shown not to act as genetic modifiers in this family. It is important to identify mutation-positive individuals and inform them of their increased risk of developing aminoglycoside induced deafness especially in a setting like South Africa where these drugs are still commonly used because of their efficacy and cost-effectiveness as a treatment for resistant forms of tuberculosis. PMID- 19285485 TI - Behavioral dissection of Drosophila larval phototaxis. AB - A behavior generally comprises multiple processes. Analyzing these processes helps to reveal more characteristics of the behavior. In this report, light/dark choice-based Drosophila larval phototaxis is analyzed with a simplistic mathematical model to reveal a fast phase and a slow phase response that are involved. Larvae of the strain w(1118), which is photophobic in phototaxis tests, prefer darkness to light in an immediate light/dark boundary passing test and demonstrate a significant reduction in motility in the dark condition during phototaxis tests. For tim(01) larvae, which show neutral performance in phototaxis tests, larvae unexpectedly prefer light to darkness in the immediate light/dark boundary passing test and demonstrate no significant motility alteration in the dark condition. It is proposed that Drosophila larval phototaxis is determined by a fast phase immediate light/dark choice and an independent slow phase light/dark-induced motility alteration that follows. PMID- 19285486 TI - CCL3/MIP-1 alpha is not involved in the LPS-induced fever and its pyrogenic activity depends on CRF. AB - The fever induced by lipopolysaccharide (LPS) depends on both prostaglandin dependent and -independent pathways. One of the prostaglandin-independent pathways is sequentially orchestrated by pre-formed pyrogenic factor derived from LPS-stimulated macrophages (PFPF), corticotrophin releasing factor (CRF), endothelin-1 (ET-1) and interleukin-1 (IL-1). As macrophage-inflammatory-protein (MIP)-1 alpha (synonym CCL3) also induces a prostaglandin independent fever, the aim of the present study was to investigate a possible participation of CCL3/MIP 1 alpha within the prostaglandin-independent pathway of LPS-induced fever which depends on PFPF, CRF and ET-1. Therefore, rats received intracerebroventricular (i.c.v.) pre-treatment with anti-CCL3 monoclonal antibody (1 and 5 ng) at 1 h and 15 min before injection of LPS (lipopolysaccharide from E. coli; 5, 50 or 100 microg kg(-1), i.v.) or CCL3/MIP-1 alpha (500 pg, i.c.v.). Both doses of anti CCL3 did not change the basal temperature but abolished the fever induced by CCL3/MIP-1 alpha. When given at the higher dose, anti-CCL3 did not influence the fever induced by i.v. injection of different doses of LPS, or i.c.v. administration of PFPF (200 ng), CRF (3 microg) or ET-1 (1 pmol). Bosentan, a non selective ET(A/B) receptors antagonist (10 microg kg(-1), i.v.), reduced the fever induced by LPS but not that induced by CCL3/MIP-1 alpha. In contrast, alpha helical CRF(9-41) (a non-selective CRF R1/R2 receptor antagonist; 25 microg injected i.c.v.) reduced CCL3/MIP-1 alpha-induced fever. In conclusion, the present results indicate that: i) CCL3/MIP-1 alpha is not an endogenous mediator of LPS-induced fever; ii) it is even not involved in the prostaglandin independent pathway of the LPS-fever cascade and iii) its pyrogenic activity depends on synthesis/release of CRF. PMID- 19285487 TI - Genetic determinants of plasma HDL-cholesterol levels in prepubertal children. AB - INTRODUCTION: Genetic determinants have been related to variation of high-density lipoprotein cholesterol (HDL-C) levels, but the extension of this association remains controversial. In our study, we analyzed the contribution of several polymorphisms on HDL-C-related genes to variation of plasma HDL-C in prepubertal children. METHODS: We studied 1269 (641 males and 628 females) 6-8 years old healthy children, who participated in a cross-sectional study examining cardiovascular risk factors in Spain. Common genetic variants in the apolipoprotein AI, apolipoprotein AII, cholesteryl ester transfer protein (CETP), hepatic lipase, ATP-binding cassette transporter A1, and paraoxonase genes were determined by PCR. RESULTS: CETP TaqI B2 carrier girls had significantly higher HDL-C levels than B1B1 girls. B2B2 boys had significantly higher (p<0.001) HDL-C than B1B1and B1B2 boys. In linear regression analysis, CETP TaqIB appears as the main predictor of HDL-C plasma levels, accounting for 4.5% and 1.8% of HDL-C variation in girls and boys respectively. CONCLUSIONS: Our data showed that among the studied polymorphisms only the CETP TaqIB polymorphism contributes to the variation in HDL-C levels in prepubertal children, particularly in girls, but overall these polymorphisms explain a small part of the variation of HDL-C plasma levels at this age. PMID- 19285489 TI - Quantitative and cell type-specific transcriptional regulation of A-type cyclin dependent kinase in Arabidopsis thaliana. AB - A-type cyclin-dependent kinase (CDKA) is an ortholog of yeast Cdc2/Cdc28p, and is assumed to have an essential function in plant growth and organogenesis. Previous studies revealed that its kinase activity is controlled by post-translational modifications, such as binding to cyclins and phosphorylations, but its transcriptional regulation is poorly understood. Here, we generated a promoter dissection series of Arabidopsis (Arabidopsis thaliana) CDKA;1, and used beta glucuronidase (GUS) gene-fused reporter constructs for expression analyses in planta. The results revealed two types of transcriptional control in shoots: general quantitative regulation and cell type-specific regulation. We identified a promoter region that promotes CDKA;1 expression in the leaf epidermis, but not in the L1 layer of the shoot apical meristem. This region also directed abaxial side-biased expression, which may be linked to the adaxial/abaxial side specification. Another reporter construct showed that CDKA;1 expression in the inner layers of leaves is controlled by a distinct regulatory region in the promoter. These results suggest that the transcriptional regulation of CDKA;1 may play a key role in proper development of leaves by coordinating cell division and differentiation of different cell types. PMID- 19285488 TI - Pleiotropic functions of embryonic sonic hedgehog expression link jaw and taste bud amplification with eye loss during cavefish evolution. AB - This study addresses the role of sonic hedgehog (shh) in increasing oral pharyngeal constructive traits (jaws and taste buds) at the expense of eyes in the blind cavefish Astyanax mexicanus. In cavefish embryos, eye primordia degenerate under the influence of hyperactive Shh signaling. In concert, cavefish show amplified jaw size and taste bud numbers as part of a change in feeding behavior. To determine whether pleiotropic effects of hyperactive Shh signaling link these regressive and constructive traits, shh expression was compared during late development of the surface-dwelling (surface fish) and cave-dwelling (cavefish) forms of Astyanax. After an initial expansion along the midline of early embryos, shh was elevated in the oral-pharyngeal region in cavefish and later was confined to taste buds. The results of shh inhibition and overexpression experiments indicate that Shh signaling has an important role in oral and taste bud development. Conditional overexpression of an injected shh transgene at specific times in development showed that taste bud amplification and eye degeneration are sensitive to shh overexpression during the same early developmental period, although taste buds are not formed until much later. Genetic crosses between cavefish and surface fish revealed an inverse relationship between eye size and jaw size/taste bud number, supporting a link between oral-pharyngeal constructive traits and eye degeneration. The results suggest that hyperactive Shh signaling increases oral and taste bud amplification in cavefish at the expense of eyes. Therefore, selection for constructive oral pharyngeal traits may be responsible for eye loss during cavefish evolution via pleiotropic function of the Shh signaling pathway. PMID- 19285490 TI - XsFRP5 modulates endodermal organogenesis in Xenopus laevis. AB - Canonical Wnt signalling is known to be involved in the regulation of differentiation and proliferation in the context of endodermal organogenesis. Wnt mediated beta-catenin activation is understood to be modulated by secreted Frizzled-related proteins, such as XsFRP5, which is dynamically expressed in the prospective liver/ventral pancreatic precursor cells during late neurula stages, becoming liver specific at tailbud stages and shifting to the posterior stomach/anterior duodenum territory during tadpole stages of Xenopus embryogenesis. These expression characteristics prompted us to analyse the function of XsFRP5 in the context of endodermal organogenesis. We demonstrate that XsFRP5 can form a complex with and inhibit a multitude of different Wnt ligands, including both canonical and non-canonical ones. Knockdown of XsFRP5 results in transient pancreatic hypoplasia as well as in an enlargement of the stomach. In VegT-injected animal cap explants, XsFRP5 can induce expression of exocrine but not endocrine pancreatic marker genes. Both, its expression characteristics as well as its interactions with XsFRP5, define Wnt2b as a putative target for XsFRP5 in vivo. Knockdown of Wnt2b results in a hypoplastic stomach as well as in hypoplasia of the pancreas. On the basis of these findings we propose that XsFRP5 exerts an early regulatory function in the specification of the ventral pancreas, as well as a late function in controlling stomach size via inhibition of Wnt signalling. PMID- 19285491 TI - Activity of new NOP receptor ligands in a rat peripheral mononeuropathy model: potentiation of morphine anti-allodynic activity by NOP receptor antagonists. AB - The effect of new NOP receptor agonists and antagonists in the rat chronic constriction injury model was investigated. Intraperitoneally administered NOP receptor agonist SR14150 and antagonists SR16430 and SR14148, had no effect on mechanical allodynia when given alone. The nonselective NOP/mu-opioid receptor agonist SR16435, however, produced an anti-allodynic response, similar to morphine and reversible by naloxone. Notably, co-administration of the NOP receptor antagonists potentiated the anti-allodynic activity of both morphine and SR16435. Increased levels of the NOP receptor are implicated in the reduced efficacy of morphine in neuropathic pain. Our results suggest the utility of NOP receptor antagonists for potentiating opioid efficacy in chronic pain. PMID- 19285492 TI - Glucosylceramide synthase inhibitors sensitise CLL cells to cytotoxic agents without reversing P-gp functional activity. AB - Malignant B-cells from most chronic lymphocytic leukaemia (CLL) patients over express MDR1 encoded P-glycoprotein (P-gp) multidrug efflux pump. Inhibition of glucosylceramide (GC) synthesis has been shown in cell lines to correlate with the expression and function of P-gp and sensitise cancer cells to cytotoxic agents. We investigated the hypothesis that reducing intracellular GC levels will reduce P-gp expression in malignant cells from CLL patients. We studied the ability of glucosylceramide synthase (GCS) inhibitors N-butyl deoxygalactonojirimycin (OGB-1) and N-nonyl-deoxygalactonojirimycin (OGB-2) to sensitise CLL cells to conventional cytotoxic drug 2-chlorodeoxyadenosine (CdA) and the cytostatic drugs chlorambucil and fludarabine. The effect on P-gp activity was analysed using the calcein-AM accumulation assay where a multidrug activity factor (MAF) of >10 in the presence of a P-gp inhibitor denotes P-gp functional activity. The P-gp over-expressing cell line CEM-VLB showed a MAF value of 96.4 with the P-gp inhibitor Z.3HCL, which fell to 15.7 after co incubation with OGB-1 and 45.9 with OGB-2. The IC(50) for vincristine fell from >10 microg/ml to 55.5 ng/ml in the presence of OGB-2. In P-gp(+ve) peripheral blood mononuclear cells from three normal volunteers, the mean MAF values for Z.3HCL, OGB-1 and OGB-2 were 23.86, 1.83 and 16.2 respectively. In 9/13 CLL samples the mean P-gp functional activity was 22.15 and P-gp was over-expressed in 12/13 samples. However, the MAF value with OGB-1 and OGB-2 was <10. Nevertheless, sensitisation in CLL cells was observed by a reduction in the IC(50) in the presence of OGB-1 and OGB-2 with the conventional drugs. We conclude that although GCS inhibitors sensitize CLL cells to cytotoxic and cytostatic drugs, they do not appear to have any effect on P-gp functional activity. PMID- 19285493 TI - Long-term nicotine treatment reduces cerebral cortical vasodilation mediated by alpha4beta2-like nicotinic acetylcholine receptors in rats. AB - Regional cortical cerebral blood flow is increased via activation of brain nicotinic acetylcholine receptors. Acute intravenous injection of nicotine increases cortical blood flow, without changing systemic blood pressure in anesthetized rats. Here, we examined whether the nicotine-induced cerebral cortical vasodilation is affected by chronic nicotine treatment. Rats received chronic subcutaneous nicotine (at a low or a high-dose) short-term (1 h) or long term (14 days). Under urethane anesthesia, blood flow in the frontal cortex, before and after bolus injection of nicotine (0.3-30 microg/kg, i.v.) was measured by laser Doppler flowmetry. The threshold dose of nicotine (3 microg/kg, i.v.) producing vasodilation was not affected by chronic nicotine treatment. However, the vasodilation induced by nicotine at 30 microg/kg was reduced after long-term nicotine treatment (but not after short-term exposure). The degree of reduction was marked and was statistically significant with high-dose (100 microg/kg/h) nicotine; low-dose (33 microg/kg/h) nicotine had a small effect that was not statistically significant. In contrast, the vasodilation in the cortical vessels obtained by hypercapnia (inhalation of 10% CO2) was not changed by chronic nicotine treatment. The nicotine-induced cortical vasodilation was not influenced by methyllycaconitine, an alpha7-selective nicotinic antagonist, while it was completely abolished by dihydro-beta-erythroidine, an alpha4beta2 preferring nicotinic antagonist. We conclude that long-term nicotine treatment reduces the functional activity of alpha4beta2-like nicotinic receptors that mediate cortical vasodilation. PMID- 19285494 TI - Proteolysis of fibrinogen deposits enables hydrogen peroxide-stimulated polymorphonuclear leukocytes to spread in an acidified environment. AB - Polymorphonuclear leukocytes might be expected to employ functional regulatory systems adapted to an acidified environment, such as found in the inflammatory sites where polymorphonuclear leukocytes act in host defense. We previously reported the unusual characteristics of phorbol 12-myristate 13-acetate (PMA) induced polymorphonuclear leukocyte spreading over immobilized fibrinogen at acidic pH, including extracellular Ca2+ requirement and independence of protein kinase C (PKC) activity. In the present study, we found that PMA-induced spreading was strongly inhibited at pH 6.0 by the serine protease inhibitor phenylmethanesulfonylfluoride at pH 6.0 but was only mildly inhibited at pH 7.2 and not inhibited at pH 8.0; furthermore, PMA-stimulated polymorphonuclear leukocytes markedly digested immobilized fibrinogen only at pH 6.0. In experiments without stimulation by PMA, we found that at pH 6.0 polymorphonuclear leukocytes were able to spread over fibrinogen surfaces pre-digested by neutrophil serine proteases; this process required extracellular Ca2+ and stimulation by hydrogen peroxide (H2O2). Pharmacological studies demonstrated the involvement of Src family protein tyrosine kinases, but not PKC, in H2O2-induced spreading over pre-digested fibrinogen surfaces; this was also the case for PMA induced spreading at pH 6.0 but not at pH 7.2 or 8.0. These results suggest that PMA-induced polymorphonuclear leukocyte spreading depends on serine protease mediated fibrinogenolysis in an acidic milieu, but that other mechanisms operate at neutral/alkaline pH. PMID- 19285495 TI - Buprenorphine inhibits bradykinin-induced release of calcitonin gene-related peptide from rat trigeminal neurons via both mu-opioid and nociceptin/orphanin peptide receptors. AB - In this study we used the dual opioid and nociceptin/orphanin peptide (NOP) agonist buprenorphine to investigate the relative contributions of opioid and NOP systems in regulating bradykinin-stimulated calcitonin-gene related peptide (CGRP) release from primary cultures of neonatal rat trigeminal neurons. We found that: bradykinin stimulates CGRP secretion either by a direct effect or after applying so-called "bradykinin-priming" protocol. In both cases, buprenorphine was able to inhibit bradykinin-stimulated CGRP secretion; however, inhibition was mediated by NOP receptors when buprenorphine was added to the incubation medium along with bradykinin, whereas it appeared to be mediated by mu-opioid receptors in bradykinin priming experiments. Bradykinin treatments also caused an increase in neuronal prostaglandin production; prostanoids appeared to be involved in the stimulatory effects of bradykinin as well as in buprenorphine inhibition, through apparently unrelated mechanisms. PMID- 19285496 TI - Neuroprotective effects of mebudipine and dibudipine on cerebral oxygen-glucose deprivation/reperfusion injury. AB - In the present study, we investigated the effects of mebudipine and dibudipine, two new Ca(2+) channel blockers, on primary murine cortical neurons exposed to oxygen-glucose deprivation/reperfusion. The experiments were performed on cells after 11-16 days of culture. To initiate oxygen-glucose deprivation /reperfusion, the culture medium was replaced by glucose-free medium, and the cells were transferred to a humidified incubation chamber in a mixture of 95% N(2) and 5% CO(2) at 37 degrees C for 30 min. The cultures were pretreated with mebudipine and dibudipine 3 h prior to oxygen-glucose deprivation/reperfusion, in order to explore their effects on neurons under oxygen-glucose deprivation conditions. Cell viability and nitric oxide (NO) production were assessed by MTT assay and the modified Griess method, respectively. Exposure of murine cortical neuronal cells to 30 min oxygen-glucose deprivation significantly decreased cell viability and increased NO production. Pretreatment of the cultures with mebudipine and dibudipine significantly increased cell viability and decreased NO generation in a dose-dependent manner. However, the drugs had no protective effect in cells subjected to oxygen-glucose deprivation for 60 min. Pretreatment of cultures with MK-801 (10 microM), a non-competitive NMDA antagonist, decreased neuronal death after 30-min oxygen-glucose deprivation, while application of NBQX (30 microM), a selective AMPA-kainate receptor antagonist, partially attenuated the cell injury. oxygen-glucose deprivation -induced cytotoxicity and NO production were also inhibited by N-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor and MK-801. We conclude that mebudipine and dibudipine could protect cortical neurons against oxygen-glucose deprivation /reperfusion-induced cell injury in a dose-dependent manner, and that this could be mediated partially by decreased NO production. PMID- 19285497 TI - Sphingosine-1-phosphate increases human alveolar epithelial IL-8 secretion, proliferation and neutrophil chemotaxis. AB - Sphingosine-1-phosphate (S1P) has been presented recently as a pro-inflammatory agent in the airway epithelium since S1P levels are increased in bronchoalveolar lavage fluid of human asthmatics. However, the effects of S1P over the alveolar epithelium and neutrophil interactions are poorly understood. Here, we show that S1P increased interleukin 8 (IL-8) gene expression and protein secretion and proliferation in alveolar epithelial cells A549 at physiological concentrations (1 microM). At the same time, S1P increased intracellular Ca2+ concentration (potency 17.91 microM, measured by epifluorescence microscopy), phospholipase D (PLD) activity (measured by chemiluminiscence method) and extracellular matrix regulated kinase1/2 (ERK1/2) phosphorylation (measured by western blot) via G(i) coupled receptor (inhibited by pertussis toxin 100 ng/ml) in A549 cells. Both, IL 8 secretion and A549 proliferation were dependent of PLD activity (inhibited by 1 butanol 0.5%), intracellular Ca2+ (inhibited by acetoxymethyl 1,2-bis(2 aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM) 100 microM), ERK1/2 phosphorylation (inhibited by 2-[2-amino-3-methoxyphenyl]-4H-1-benzopyran-4-one (PD98059) 10 microM) and G(i)-coupled receptors (blocked by pertussis toxin 100 ng/ml). Moreover, S1P increased intercellular adhesion molecule I (ICAM-1) expression and failed in vascular cell adhesion molecule I (VCAM-1) modification (measured by flow cytometer) in A549. Indirectly, A549 supernatant fluids arising from A549-S1P 1 microM stimulation decreased L-selectin expression without CD11b/CD18 integrin modification in human neutrophils. In the same way, A549-S1P supernatant fluids increased neutrophil chemotaxis (Boyden chamber), which was inhibited by antibody against IL-8. This study demonstrates for the first time that S1P participates in the alveolar epithelial interactions in vitro. PMID- 19285498 TI - Gabapentin and pregabalin ameliorate mechanical hypersensitivity after spinal cord injury in mice. AB - The antiepileptic drugs gabapentin and pregabalin exhibit well-established analgesic effects in patients with several neuropathic conditions. In the present study, we examined their effects on mechanical hypersensitivity in mice subjected to weight-drop spinal cord injury. Hindlimb motor function and mechanical hypersensitivity were evaluated using the Basso-Beattie-Bresnahan (BBB) locomotor rating scale and the von Frey test, respectively, for 4 weeks after spinal cord injury. Despite gradual recovery of hindlimb motor function after spinal cord injury, mice exhibited continuous development of mechanical hypersensitivity. Gabapentin (30 and 100 mg/kg) and pregabalin (10 and 30 mg/kg), administered intraperitoneally on the 28th day after spinal cord injury, reduced mechanical hypersensitivity in a dose-dependent manner. These results suggest that gabapentin and pregabalin could be useful therapeutic tools for patients with neuropathic pain after spinal cord injury. PMID- 19285499 TI - Corneal myofibroblast viability: opposing effects of IL-1 and TGF beta1. AB - The purpose of this study was to test the effect of corneal epithelial scrape on myofibroblasts associated with haze and elucidate the effect of interleukin-1 and transforming growth factor beta1 on corneal stromal myofibroblasts viability and death in vitro. Corneal epithelial scrape was performed in rabbit eyes with severe haze at one month after -9 diopter photorefractive keratectomy. Corneas were processed for immunocytochemistry for myofibroblast marker alpha-smooth muscle actin (alpha-SMA) and the TUNEL assay to detect apoptosis. Rabbit corneal fibroblasts were cultured with 2 ng/ml of transforming growth factor beta1 (TGF beta1) to induce myofibroblast differentiation confirmed by monitoring alpha-SMA expression. Fluorescence-based TUNEL assay was performed to analyze the apoptotic response of myofibroblasts to IL-1alpha or IL-1beta, in the presence or absence of TGF beta1. Dose response experiments were performed after withdrawal of TGF beta1 and exposure to 1, 5, or 10 ng/ml of IL-1alpha or IL-1beta for 1 h. Subsequent experiments were performed with myofibroblasts exposed to 5 ng/ml of IL-1alpha or IL-1beta in conjunction with 0, 1, 5, or 10 ng/ml of TGF beta1. Corneal epithelial scrape with a scalpel blade produced myofibroblast apoptosis. Exposure to TGF beta1 in vitro resulted in greater than 99% transformation of corneal fibroblasts to alpha-SMA+ myofibroblasts. There was a statistically significant dose-dependent increase in the percentage of TUNEL+ cells with either IL-1alpha or IL-1beta initiated at concentrations as low as 1 ng/ml. For example, after withdrawal of TGF beta1, the % TUNEL+ cells at 1 h after exposure to IL 1alpha increased significantly with increasing concentration (0 ng/ml, 2.4 +/- 0.8% [S.E.M.]; 1 ng/ml, 15.4 +/- 1.8%; 5 ng/ml, 47.4 +/- 3.9%; or 10 ng/ml, 70.3 +/- 3.2%). Similar results were obtained with IL-1beta. The differences between the means of apoptotic myofibroblasts for the different concentrations of cytokine for either IL-1alpha or IL-1beta were significantly different (ANOVA, p < 0.001). When myofibroblasts were exposed to 5 ng/ml of IL-1alpha or IL-1beta, the % TUNEL+ cells at 1 h were reduced in a significant dose-dependent manner when TGF beta1 at a concentration of 5 ng/ml or 10 ng/ml was present in the medium (ANOVA p < 0.01). IL-1alpha or IL-1beta triggers the death of myofibroblasts in vitro and TGF beta1 reduces the IL-1 effect on cell death. TGF beta1 and IL-1 have opposing effects on myofibroblast viability and likely interact to modulate haze generation after corneal injury. PMID- 19285500 TI - The amino-terminal region of Atg3 is essential for association with phosphatidylethanolamine in Atg8 lipidation. AB - Autophagy is a bulk degradation process conserved among eukaryotes. In macro autophagy, autophagosomes sequester cytoplasmic components and deliver their contents to lysosomes/vacuoles. Autophagosome formation requires the conjugation of Atg8, a ubiquitin-like protein, to phosphatidylethanolamine (PE). Here we report that the amino (N)-terminal region of Atg3, an E2-like enzyme for Atg8, plays a crucial role in Atg8-PE conjugation. The conjugating activities of Atg3 mutants lacking the 7 N-terminal amino acid residues or containing a Leu-to-Asp mutation at position 6 were severely impaired both in vivo and in vitro. In addition, the amino-terminal region is critical for interaction with the substrate, PE. PMID- 19285501 TI - Analysis of the translationally controlled tumour protein in the nematodes Ostertagia ostertagi and Caenorhabditis elegans suggests a pivotal role in egg production. AB - The translationally controlled tumour protein (TCTP) is a conserved protein which has been described for a wide range of eukaryotic organisms including protozoa, yeasts, plants, nematodes and mammals. Several parasitic organisms have been shown to actively secrete TCTP during host infection as part of their immuno evasive strategy. In this study, we have studied TCTP in Ostertagia ostertagi, a parasitic nematode of cattle, and in the free-living nematode Caenorhabditis elegans. An analysis of the transcription and expression patterns showed that TCTP was present in the eggs of both species. This localisation is consistent for some other Strongylida such as Teladorsagia circumcincta, Cooperia oncophora and Haemonchus contortus. TCTP was also detected at low levels in excretory-secretory material from adult O. ostertagi worms. The role of TCTP in nematode biology was also investigated by RNA interference in C. elegans. Knock-down of C. elegans tctp (tct-1) transcription reduced the numbers of eggs laid by the hermaphrodite in the F(0) and F(1) generations by 90% and 72%, respectively, indicating a pivotal role of TCTP in reproduction. PMID- 19285502 TI - Schistosoma haematobium total antigen induces increased proliferation, migration and invasion, and decreases apoptosis of normal epithelial cells. AB - Schistosome worms are blood-dwelling flukes that cause chronic infection in more than 200 million people and are thought to be responsible for 500,000 deaths annually. During infection with Schistosoma haematobium, eggs are deposited in the mucosa and submucosa of the bladder and lower ureters. Squamous cell carcinoma (SCC) of the bladder is a long-term sequela of chronic infection. The mechanisms underlying the association between S. haematobium and SCC of the bladder are largely unknown, with all reports to date exclusively demonstrating epidemiological evidence linking S. haematobium infection with SCC of the bladder. We hypothesised that the parasite antigens might induce alterations in epithelial cells towards cancer. For this we used Chinese Hamster Ovary (CHO) cells and treated the cells in culture with S. haematobium total antigen (Sh). Our results showed increased proliferation, increased S-phase and decreased apoptosis, as well as down-regulation of tumour suppressor p27 and up-regulation of anti-apoptotic molecule Bcl-2 in Sh-treated cells compared with controls. We also found increased migration and invasion. To our knowledge, this is the first report demonstrating alterations of normal epithelial cells as a direct effect of S. haematobium antigens. PMID- 19285503 TI - A novel Salifa-Saprolegnia association. AB - During the period from December 2006 to March 2007, about 1000 freshwater leeches, Salifa delicata, were collected from Al-sont canal, adjacent to Assiut city, Egypt. In the laboratory, 96% of Salifa delicata showed signs of oomycotal infection (cotton-wool like appearance radiating out in whorled pattern) and died within 3 days. Direct microscopy and culture proved Saprolegnia hypogyna to be the pathogen. Histopathological studies showed necrotic lesions, destruction of cuticle, epidermis, dermis, muscle layers, botryoidal tissue and even the gut with the oomycete hyphae penetrating the damaged tissues. To the best of the authors' knowledge, this represents the first record of a novel association between the leech, Salifa delicata, and the oomycete, Saprolegnia hypogyna, but the second report on the histopathology of saprolegniasis within leeches. PMID- 19285504 TI - Mitochondrial calcium transport in the heart: physiological and pathological roles. AB - That intramitochondrial free calcium ([Ca(2+)](m)) plays various critical roles in both normal physiological and pathological conditions in the heart is now well accepted, and evidenced by the interest and work in this area of the last two decades. However, controversies remain; such as the existence of beat-to-beat mitochondrial Ca(2+) transients, role of [Ca(2+)](m) in modulating whole-cell Ca(2+) signalling, whether or not [Ca(2+)](m) is critical for increases in ATP supply upon increased demand, and its role in cell death by both necrosis and apoptosis, especially in formation of the mitochondrial permeability transition pore and in ischaemic preconditioning. Neither is there a consensus as to whether inhibiting the Ca(2+) influx or efflux pathways--the Ca(2+) uniporter (MCU) and Na(+)/Ca(2+)-excahnger (mNCX), respectively--is cardioprotective, largely due to lack of specific inhibitors of these transporters. Ruthenium red, Ru360, clonazepam and CGP37157 are all very effective in isolated mitochondria, but reports of their effectiveness in whole cell and heart studies vary considerably, which partly accounts for the lack of a consensus on protective effects. The purification and cloning of the transporters, and development of more specific inhibitors, would produce a step-change in our understanding of the role of these apparently critical but still elusive proteins. However, developments in fluorescent indicators, proteins and imaging technology have meant that [Ca(2+)](m) can now be measured reasonably specifically in intact cells and hearts, and interactions of the mitochondrial Ca(2+) transporters with those of the sarcolemma or sarcoplasmic reticulum are being revealed. This has gone a long way to bringing the transporters to the forefront of cardiac research. PMID- 19285505 TI - Structural rearrangements in the active site of the Thermus thermophilus 16S rRNA methyltransferase KsgA in a binary complex with 5'-methylthioadenosine. AB - Posttranscriptional modification of ribosomal RNA (rRNA) occurs in all kingdoms of life. The S-adenosyl-L-methionine-dependent methyltransferase KsgA introduces the most highly conserved rRNA modification, the dimethylation of A1518 and A1519 of 16S rRNA. Loss of this dimethylation confers resistance to the antibiotic kasugamycin. Here, we report biochemical studies and high-resolution crystal structures of KsgA from Thermus thermophilus. Methylation of 30S ribosomal subunits by T. thermophilus KsgA is more efficient at low concentrations of magnesium ions, suggesting that partially unfolded RNA is the preferred substrate. The overall structure is similar to that of other methyltransferases but contains an additional alpha-helix in a novel N-terminal extension. Comparison of the apoenzyme with complex structures with 5'-methylthioadenosine or adenosine bound in the cofactor-binding site reveals novel features when compared with related enzymes. Several mobile loop regions that restrict access to the cofactor-binding site are observed. In addition, the orientation of residues in the substrate-binding site indicates that conformational changes are required for binding two adjacent residues of the substrate rRNA. PMID- 19285506 TI - Structure of the third intracellular loop of the vasopressin V2 receptor and conformational changes upon binding to gC1qR. AB - The V2 vasopressin receptor is a G-protein-coupled receptor that regulates the renal antidiuretic response. Its third intracellular loop is involved in the coupling not only with the GalphaS protein but also with gC1qR, a potential chaperone of G-protein-coupled receptors. In this report, we describe the NMR solution structure of the V2 i3 loop under a cyclized form (i3_cyc) and characterize its interaction with gC1qR. i3_cyc formed a left-twisted alpha helical hairpin structure. The building of a model of the entire V2 receptor including the i3_cyc NMR structure clarified the side-chain orientation of charged residues, in agreement with literature mutagenesis reports. In the model, the i3 loop formed a rigid helical column, protruding deep inside the cytoplasm, as does the i3 loop in the recently elucidated structure of squid rhodopsin. However, its higher packing angle resulted in a different structural motif at the intracellular interface, which may be important for the specific recognition of GalphaS. Moreover, we could estimate the apparent K(d) of the i3_cyc/gC1qR complex by anisotropy fluorescence. Using a shorter and more soluble version of i3_cyc, which encompassed the putative site of gC1qR binding, we showed by NMR saturation transfer difference spectroscopy that the binding surface corresponded to the central arginine cluster. Binding to gC1qR induced the folding of the otherwise disordered short peptide into a spiral-like path formed by a succession of I and IV turns. Our simulations suggested that this folding would rigidify the arginine cluster in the entire i3 loop and would alter the conformation of the cytosolic extensions of TM V and TM VI helices. In agreement with this conformational rearrangement, we observed that binding of gC1qR to the full length receptor modifies the intrinsic tryptophan fluorescence binding curves of V2 to an antagonist. PMID- 19285507 TI - Indirect DNA readout on the protein side: coupling between histidine protonation, global structural cooperativity, dynamics, and DNA binding of the human papillomavirus type 16 E2C domain. AB - DNA sequence recognition by the homodimeric C-terminal domain of the human papillomavirus type 16 E2 protein (E2C) is known to involve both direct readout and DNA-dependent indirect readout mechanisms, while protein-dependent indirect readout has been deduced but not directly observed. We have investigated coupling between specific DNA binding and the dynamics of the unusual E2C fold, using pH as an external variable. Nuclear magnetic resonance and isothermal titration calorimetry show that pH titration of His318 in the complex interface and His288 in the core of the domain is coupled to both binding and the dynamics of the beta barrel core of E2C, with a tradeoff between dimer stability and function. Specific DNA binding is, in turn, coupled to the slow dynamics and amide hydrogen exchange in the entire beta-barrel, reaching residues far apart from the DNA recognition elements but not affecting the two helices of each monomer. The changes are largest in the dimerization interface, suggesting that the E2C beta barrel acts as a hinge that regulates the relative position of the DNA recognition helices. In conclusion, the cooperative dynamics of the human papillomavirus type 16 E2C beta-barrel is coupled to sequence recognition in a protein-dependent indirect readout mechanism. The patterns of residue substitution in genital papillomaviruses support the importance of the protonation states of His288 and His318 and suggest that protein-dependent indirect readout and histidine pH titration may regulate DNA binding in the cell. PMID- 19285508 TI - Blood group antigen recognition by a solute-binding protein from a serotype 3 strain of Streptococcus pneumoniae. AB - Streptococcus pneumoniae is a common bacterial pathogen that is well known for its ability to cause acute respiratory disease (pneumonia), ear infections, and other serious illnesses. This Gram-positive bacterium relies on its carbohydrate metabolizing capabilities for full virulence in its host; however, the range of glycan targets that it can attack is presently not fully appreciated. S. pneumoniae is known to have a fucose utilization operon that in the TIGR4 strain plays a role in its virulence. Here we identify a second type of fucose utilization operon that is present in a subset of S. pneumoniae strains, including the serotype 3 strain SP3-BS71. This operon contains a transporter with a solute-binding protein, FcsSBP (fucose solute-binding protein), that interacts tightly (Ka approximately 1 x 10(6) M(-1)) and specifically with soluble A- and B antigen trisaccharides but displays no selectivity between these two sugars. The structure of the FcsSBP in complex with the A-trisaccharide antigen, determined to 2.35 A, reveals its mode of binding to the reducing end of this sugar, thus highlighting this protein's requirement for soluble blood group antigen ligands. Overall, this report exposes a heretofore unknown capability of certain S. pneumoniae strains to transport and potentially metabolize the histo-blood group antigen carbohydrates of its host. PMID- 19285509 TI - Evolving learning rules and emergence of cooperation in spatial prisoner's dilemma. AB - In the evolutionary Prisoner's dilemma (PD) game, agents play with each other and update their strategies in every generation according to some microscopic dynamical rule. In its spatial version, agents do not play with every other but, instead, interact only with their neighbours, thus mimicking the existing of a social or contact network that defines who interacts with whom. In this work, we explore evolutionary, spatial PD systems consisting of two types of agents, each with a certain update (reproduction, learning) rule. We investigate two different scenarios: in the first case, update rules remain fixed for the entire evolution of the system; in the second case, agents update both strategy and update rule in every generation. We show that in a well-mixed population the evolutionary outcome is always full defection. We subsequently focus on two-strategy competition with nearest-neighbour interactions on the contact network and synchronised update of strategies. Our results show that, for an important range of the parameters of the game, the final state of the system is largely different from that arising from the usual setup of a single, fixed dynamical rule. Furthermore, the results are also very different if update rules are fixed or evolve with the strategies. In these respect, we have studied representative update rules, finding that some of them may become extinct while others prevail. We describe the new and rich variety of final outcomes that arise from this co evolutionary dynamics. We include examples of other neighbourhoods and asynchronous updating that confirm the robustness of our conclusions. Our results pave the way to an evolutionary rationale for modelling social interactions through game theory with a preferred set of update rules. PMID- 19285510 TI - Survival benefits in mimicry: a quantitative framework. AB - Mimicry is a resemblance between species that benefits at least one of the species. It is a ubiquitous evolutionary phenomenon particularly common among prey species, in which case the advantage involves better protection from predation. We formulate a mathematical description of predation, to investigate benefits and disadvantages of mimicry. The basic setup involves differential equations for quantities representing predator behavior, namely, the probabilities for attacking prey at the next encounter. Using this framework, we present new quantitative results, and also provide a unified description of a significant fraction of the quantitative mimicry literature. The new results include "temporary" mutualism between prey species, and an optimal density at which the survival benefit is greatest for the mimic. The formalism leads naturally to extensions in several directions, such as the interplay of mimicry with population dynamics, studies of spatiotemporal patterns, etc. We demonstrate this extensibility by presenting some explorations on spatiotemporal pattern dynamics. PMID- 19285511 TI - The shape of the competition and carrying capacity kernels affects the likelihood of disruptive selection. AB - Many quantitative genetic and adaptive dynamic models suggest that disruptive selection can maintain genetic polymorphism and be the driving force causing evolutionary divergence. These models also suggest that disruptive selection arises from frequency-dependent intraspecific competition. For convenience or historical precedence, these models assume that carrying capacity and competition functions follow a Gaussian distribution. Here, we propose a new analytical framework that relaxes the assumption of Gaussian competition and carrying capacity functions, and investigate how alternative shapes affect the likelihood of disruptive selection. We found that the shape of both carrying capacity and competition kernels interact to determine the likelihood of disruptive selection. For certain regions of the parametric space disruptive selection is facilitated, whereas for others it becomes more difficult. Our results suggest that the relationship between the degree of frequency dependence and the likelihood of disruptive selection is more complex than previously thought, depending on how resources are distributed and competition interference takes place. It is now important to describe the empirical patterns of resource distribution and competition in nature as a way to determine the likelihood of disruptive selection in natural populations. PMID- 19285512 TI - Dynamic energy budgets in syntrophic symbiotic relationships between heterotrophic hosts and photoautotrophic symbionts. AB - In this paper we develop and investigate a dynamic energy budget (DEB) model describing the syntrophic symbiotic relationship between a heterotrophic host and an internal photoautotrophic symbiont. The model specifies the flows of matter and energy among host, symbiont and environment with minimal complexity and uses the concept of synthesizing units to describe smoothly the assimilation of multiple limiting factors, in particular inorganic carbon and nitrogen, and irradiance. The model has two passive regulation mechanisms: the symbiont shares only photosynthate that it cannot use itself, and the host delivers only excess nutrients to the symbiont. With parameter values plausible for scleractinian corals, we show that these two regulation mechanisms suffice to obtain a stable symbiotic relationship under constant ambient conditions, provided those conditions support sustenance of host and symbiont. Furthermore, the symbiont density in the host varies relatively little as a function of ambient food density, inorganic nitrogen and irradiance. This symbiont density tends to increase with light deprivation or nitrogen enrichment, either directly or via food. We also investigate the relative benefit each partner derives from the relationship and conclude that this relationship may shift from mutualism to parasitism as environmental conditions change. PMID- 19285513 TI - Evolution of cell motility in an individual-based model of tumour growth. AB - Tumour invasion is driven by proliferation and importantly migration into the surrounding tissue. Cancer cell motility is also critical in the formation of metastases and is therefore a fundamental issue in cancer research. In this paper we investigate the emergence of cancer cell motility in an evolving tumour population using an individual-based modelling approach. In this model of tumour growth each cell is equipped with a micro-environment response network that determines the behaviour or phenotype of the cell based on the local environment. The response network is modelled using a feed-forward neural network, which is subject to mutations when the cells divide. With this model we have investigated the impact of the micro-environment on the emergence of a motile invasive phenotype. The results show that when a motile phenotype emerges the dynamics of the model are radically changed and we observe faster growing tumours exhibiting diffuse morphologies. Further we observe that the emergence of a motile subclone can occur in a wide range of micro-environmental growth conditions. Iterated simulations showed that in identical growth conditions the evolutionary dynamics either converge to a proliferating or migratory phenotype, which suggests that the introduction of cell motility into the model changes the shape of fitness landscape on which the cancer cell population evolves and that it now contains several local maxima. This could have important implications for cancer treatments which focus on the gene level, as our results show that several distinct genotypes and critically distinct phenotypes can emerge and become dominant in the same micro-environment. PMID- 19285514 TI - Energetic analysis of the two controversial drug binding sites of the M2 proton channel in influenza A virus. AB - Understanding the mechanism of the M2 proton channel of influenza A is crucially important to both basic research and drug discovery. Recently, the structure was determined independently by high-resolution NMR and X-ray crystallography. However, the two studies lead to completely different drug-binding mechanisms: the X-ray structure shows the drug blocking the pore from inside; whereas the NMR structure shows the drug inhibiting the channel from outside by an allosteric mechanism. Which one of the two is correct? To address this problem, we conducted an in-depth computational analysis. The conclusions drawn from various aspects, such as energetics, the channel-gating dynamic process, the pK(a) shift and its impact on the channel, and the consistency with the previous functional studies, among others, are all in favour to the allosteric mechanism revealed by the NMR structure. The findings reported here may stimulate and encourage new strategies for developing effective drugs against influenza A, particularly in dealing with the drug-resistant problems. PMID- 19285515 TI - Signaling pathways associated with the expression of inflammatory mediators activated during the course of two models of experimental periodontitis. AB - AIMS: Evaluate the signaling pathways associated with inflammatory mediators activated in two models of experimental periodontitis. MAIN METHODS: Two models were used: lipopolysaccharide (LPS) injections and ligature placement. Wistar rats were used and 30 microg LPS from Escherichia coli was injected twice a week into the palatal aspect of the upper molars. Ligatures were placed around lower first molars. A control group received injections of PBS on the palatal gingivae whereas no ligatures were placed on the lower molars. Samples were collected 5, 15 and 30 days and processed for analysis by Western blotting and stereometry. KEY FINDINGS: The ligature model was associated with rapid and transient activation of extracellular-regulated kinases (ERK) and p38 mitogen-activated protein kinase (MAPK) as well as of nuclear factor kappa B (NF-kappaB). Activation of these signaling pathways on the LPS model was delayed but sustained throughout the 30-day experimental period. Inflammatory changes induced by both models were similar; however there was a significant reduction on inflammation degree on the ligature model, which paralleled the decrease observed on the activation of the signaling pathways. Activation of signal transducer and activator of transcription (STAT)-3 by phosphorylation of Tyrosine residues and of STAT-5 was observed only on the ligature model. SIGNIFICANCE: Regulation of gene expression results from the activation of signaling pathways initiated by receptor-ligand binding of external antigens and also of cytokines produced by the host immune system. Understanding the signaling pathways relevant for a given condition may provide information useful for novel therapeutic approaches. PMID- 19285516 TI - Analysis of protein synthesis dynamic model in eukaryotic cells: Input control. AB - This paper presents the analysis of initiation control model of protein synthesis via eukaryotic initiation factor (eIF)-2 unit, introduced by [N.S. Bar, D.R. Morris, Dynamic model of the process of protein synthesis in eukaryoric cells, Bulletin of Mathematical Biology 69 (2007) 361-393, doi:10.1007/s11538-006-9128 2.] and propose methods to control it. Linearization of the model is presented as a measure to simplify the analysis and control application. The properties of the linear model were investigated and compared to the non-linear model using simulations. It was shown that the linear model is (marginally) stable and the states converge to a finite value. Linear optimal control theory can then be applied to the model under the value range where the linearized model is accurate. The effect of the input signals GCN2.tRNA and eIF-2 on the non-linear system was investigated. A few characteristics known from in vitro experiments of the initiation process were proven from a mathematical aspect and some conclusions about the function of the initiation complexes such as eIF2B and the ternary complex were derived. Consistent with published experiments, it was shown that overexpression of eIF-2 increases the concentration of 48S initiation complex and promote initiation rate. A state feedback control was applied in order to manipulate the initiation rate and it was proven that the 48S initiation complex can be driven to a desired value by calculating an input control law using measurement techniques available today. If this strategy can be implemented de facto, then a genuine control on protein synthesis process can be obtained. PMID- 19285517 TI - Effects of N-acetylaspartylglutamate (NAAG) at group II mGluRs and NMDAR. AB - A group II metabotropic glutamate receptor (mGluR) agonist was recently reported to be clinically efficacious against symptoms of schizophrenia [Patil, S.T., Zhang, L., Martenyi, F., Lowe, S.L., Jackson, K.A., Andreev, B.V., Avedisova, A.S., Bardenstein, L.M., Gurovich, I.Y., Morozova, M.A., Mosolov, S.N., Neznanov, N.G., Reznik, A.M., Smulevich, A.B., Tochilov, V.A., Johnson, B.G., Monn, J.A., Schoepp, D.D., 2007. Activation of mGlu2/3 receptors as a new approach to treat schizophrenia: a randomized phase 2 clinical trial. Nature Med 13, 1102-1107]. The endogenous neuropeptide N-acetylaspartylglutamate (NAAG) has been described as an agonist at mGluR2 and mGluR3 [Wroblewska, B., Wroblewski, J.T., Pshenichkin, S., Surin, A., Sullivan, S.E., Neale, J.H., 1997. N acetylaspartylglutamate selectively activates mGluR3 receptors in transfected cells. J. Neurochem. 69, 174-181; Cartmell, J., Adam, G., Chaboz, S., Henningsen, R., Kemp, J.A., Klingelschmidt, A., Metzler, V., Monsma, F., Schaffhauser, H., Wichmann, J., Mutel, V., 1998. Characterization of [3H]-(2S,2'R,3'R)-2-(2',3' dicarboxy-cyclopropyl)glycine ([3H]-DCG IV) binding to metabotropic mGlu2 receptor-transfected cell membranes. Br. J. Pharmacol. 123, 497-504] and is degraded by the enzyme glutamate carboxypeptidase II (also known as N-acetyl alpha-linked acidic dipeptidase or NAALADase). Hence, elevating the concentration of endogenous NAAG by inhibition of NAALADase represents a potential strategy for the treatment of schizophrenia via group II mGluR activation. We therefore investigated the activity of NAAG at both rat native and human recombinant mGluRs. We found that NAAG had no effect on synaptic transmission at the medial perforant pathway inputs to the rat dentate gyrus which is known to be sensitive to group II mGluR activation. We proceeded to examine the effects of NAAG at human recombinant mGluR2 and mGluR3 in a cellular G protein-activated K+ channel electrophysiology assay. Furthermore, due to discrepancies in the literature concerning the activity of NAAG at the N-methyl-d-aspartate receptor [NMDAR; Westbrook, G.L., Mayer, M.L., Namboodiri, M.A., Neale, J.H., 1986. High concentrations of N-acetylaspartylglutamate (NAAG) selectively activate NMDA receptors on mouse spinal cord neurons in cell culture. J. Neurosci. 6, 3385 3392; Losi, G., Vicini, S., Neale, J., 2004. NAAG fails to antagonize synaptic and extrasynaptic NMDA receptors in cerebellar granule neurons. Neuropharmacology 46, 490-496], we also tested NAAG at NMDARs in rat hippocampal neurons in culture. We found that a purified NAAG preparation had no effect at mGluR2, mGluR3 or NMDAR. Taken together, these findings do not support a rationale for targeting NAALADase and increasing extracellular NAAG levels as a therapeutic strategy for the treatment of schizophrenia. PMID- 19285519 TI - Poisoning by Centratherum brachylepis in ruminants. AB - A disease causing anorexia and ruminal indigestion in cattle and goats, and also edema of the lips, tongue and face in goats, was associated with the ingestion of Centratherum brachylepis in pastures containing large amounts of the plant. On 3 farms with a total of 217 cattle and 140 goats, 57 (26%) cattle and 56 (40%) goats were affected, and 11 (5%) cattle and 34 (24%) goats died. In one cow that died there were widespread and severe histologic lesions in the rumen that consisted of vacuolation and ballooning degeneration of keratinocytes, and vesicle and pustule formation in the epithelium. C. brachylepis was administered orally to 3 sheep and 11 goats. Clinical signs similar to those observed in spontaneous field cases in goats were reproduced in 2 sheep and 3 goats that ingested 30-50 g/kg body weight of the plant when administered within 48 h of it being collected. C. brachylepis collected between 2 and 13 days prior to being administered caused no clinical signs in 1 sheep and 8 goats at dose rates of 30 300 g/kg body weight of the plant. These feeding studies provide evidence that C. brachylepis is the cause of the field disease observed and that the plant loses toxicity after harvesting. PMID- 19285518 TI - Dehydromonocrotaline induces cyclosporine A-insensitive mitochondrial permeability transition/cytochrome c release. AB - Monocrotaline (MCT) is a pyrrolizidine alkaloid present in plants of the genus Crotalaria that causes cytotoxicity and genotoxicity in animals and humans. It is well established that the toxicity of MCT results from its hepatic bioactivation to dehydromonocrotaline (DHM), an alkylating agent, but the exact mechanism of action remains unknown. In a previous study, we demonstrated DHM's inhibition of mitochondrial NADH-dehydrogenase activity at micromolar concentrations, which is an effect associated with a significant reduction in ATP synthesis. As a follow up study, we have evaluated the ability of DHM to induce mitochondrial permeability transition (MPT) and its associated processes in isolated rat liver mitochondria. In the presence of 10 microM Ca(2+), DHM (50-250 microM) elicited MPT in a concentration-dependent, but cyclosporine A-independent manner, as assessed by mitochondrial swelling, which is associated with mitochondrial Ca(2+) efflux and cytochrome c release. DHM (50-250 microM) did not cause hydrogen peroxide accumulation but did deplete endogenous glutathione and NAD(P)H, while oxidizing protein thiol groups. These results potentially indicate the involvement of mitochondria, via apoptosis, in the well-documented cytotoxicity of monocrotaline. PMID- 19285520 TI - Venom from the centipede Scolopendra viridis Say: purification, gene cloning and phylogenetic analysis of a phospholipase A2. AB - Venom components from the centipede Scolopendra viridis Say were studied, using both the soluble venom and a cDNA library prepared from mRNA of the venomous glands. Separation of the soluble venom by high performance liquid chromatography (HPLC) permitted to obtain at least 54 different fractions. The fraction eluting at 46.24 min showed phospholipase activity. The enzyme was purified to homogeneity and the first 25 amino acid residues were identified by Edman degradation. From the cDNA library several genes were cloned, one of which codes for a protein with identical amino acid sequence as the one experimentally determined. The cloned gene codes for a signal peptide of 28 amino acids and a mature peptide of 119 residues. The molecular weight of the enzyme was estimated by mass spectrometry and shown to be 13,752 Da, which matches exactly with the molecular mass expected from the deduced amino acid sequence of the gene. Phylogenetic analysis of this sequence, in comparison with other known from venomous animals, showed that it is more similar to snake phospholipases than to insect or arachnid sequences, suggesting that it has been submitted to convergent evolution. To the best of our knowledge this is the first time that a phospholipase from this species of animal is fully characterized. We have named it Scol/Pla. PMID- 19285521 TI - Effect of averageness and sexual dimorphism on the judgment of facial attractiveness. AB - Effect of sexual dimorphism and averageness on the judgment of facial attractiveness was investigated. Participants (n=114) rated attractiveness of 96 facial photographs with neutral expressions. Principal component analyses were conducted on 80 facial feature points standardized via the generalized Procrustes method. Local regression analysis was used to obtain the distribution of attractiveness evaluations for the first two principal components. The distribution of facial attractiveness of each sex was approximately line symmetrical, and each axis of the symmetry passed through average male and female faces. These results suggest that sexual dimorphism and averageness independently influence facial attractiveness. PMID- 19285522 TI - Interpretation of optic flows synchronized with observer's hand movements. AB - We investigated whether the visual system could use a novel action-perception relationship mediated by a touch panel to resolve ambiguity in 2D optic flows. The stimulus was an optic flow produced by a dotted plane, which was translated and rotated in depth. The translation was synchronized with subject's hand movements on a touch panel. There were two perceptual interpretations of the stimulus as a surface patch oriented in 3D: (1) approaching it in depth and rotating away from gaze normal, or (2) not translating it in depth and rotating toward gaze normal around an axis perpendicular to that of Case 1 [Wexler, M., Lamouret, I., & Droulez, J. (2001a). The stationarity hypothesis: an allocentric criterion in visual perception. Vision Research, 41, 3023-3037]. Subjects reported the direction of the axis of rotation, which was perceptually coupled with the perception of translation in depth. The results indicate that the frequency of perception in Case 1 increased as the sessions progressed. This suggests that the visual system learned the association between hand movements and viewpoint translation during the experiment and used this association to decompose the optic flow. PMID- 19285523 TI - The perception of speed based on L-M and S-(L+M) cone opponent processing. AB - We have measured perceived speed and speed discrimination thresholds for stimuli that selectively activate the L-M, S-(L+M) cone opponent and L+M (luminance) post receptoral pathways. For low speeds and low contrasts speed discrimination thresholds for L-M and S-(L+M) are similar but are higher and have a greater dependency upon contrast than those for luminance motion. These differences between chromatic and luminance speed perception can be eliminated when stimuli are equated with respect to their individual motion detection thresholds (MDTs). For fast moving gratings speed perception based upon L-M, S-(L+M) and L+M signals is similar in terms of threshold performance and contrast dependency. These results are consistent with the view that there are separate mechanisms for the analysis of chromatic and luminance motion, the relative contributions of which may change as a function of stimulus contrast and speed. The similarity in performance for S-(L+M) and L+M chromatic stimuli across a range of stimulus parameters suggests that signals derived from the two cone opponent pathways can be used equally well. Our results argue against the idea that speed perception is compromised when it is based upon information derived from the S-(L+M) cone opponent pathway. PMID- 19285524 TI - Factors associated with response to a follow-up postal questionnaire in a cohort of American Indians. AB - OBJECTIVE: The aim of the study was to collect pilot data on response rates to a follow-up postal questionnaire in a cohort of American Indians living in the Southwestern United States. We tested the effect of questionnaire length on response. METHODS: Cohort members were American Indian adults aged 18 and over who completed the baseline study visit. Study participants (N=1587), cohort members who completed the baseline study visit during the first year of enrollment, were randomized into two groups to receive either an 18-page or 3 page follow-up postal questionnaire. Data were collected between October 2005 and March 2006. RESULTS: The response rates after two questionnaire mailings and a reminder postcard were significantly higher for the short versus the long (56.2%, 48.1% p= < 0.01) questionnaire. Being female and being aged 50 or older were associated with returning a completed questionnaire. A reminder postcard and second mailing improved response by 11.7% and 13.4% respectively. CONCLUSIONS: These results show that a postal questionnaire can be used in a cohort of American Indians living in the Southwest, but suggest that questionnaires should be short and repeat mailings are needed. PMID- 19285525 TI - Perceived enforcement of school tobacco policy and adolescents' cigarette smoking. AB - OBJECTIVES: School tobacco use policies are part of a comprehensive strategy for preventing or reducing adolescent cigarette smoking. This study examines the relationship between perceived tobacco policy enforcement at the school level and smoking behaviors among students. METHODS: 21,281 middle and high school students of 255 schools participated in the 2006 Oregon Health Teens Survey. Multilevel logistic regression was conducted, using a school-level policy enforcement measure based on aggregated student reports, and individual-level characteristics (e.g., age, gender, cigarette smoking before age 12, personal beliefs about smoking) as predictors of past-30-day cigarette smoking behaviors (e.g., any smoking, daily smoking, heavy episodic smoking, smoking on school property). RESULTS: Higher levels of perceived enforcement of anti-smoking policy at the school level were inversely associated with the prevalence of past-30-day smoking behaviors, independent of individual-level predictors. CONCLUSIONS: Stricter enforcement of school policies against tobacco use may help prevent or reduce adolescents' cigarette smoking on and outside of school property. PMID- 19285526 TI - Prostanoids and reactive oxygen species: team players in endothelium-dependent contractions. AB - The endothelial cells control the tone of the underlying vascular smooth muscle by releasing vasoactive substances. Endothelium-derived relaxing factors (EDRF), in particular nitric oxide have received considerable attention, but much less is known about the ability of the endothelial cells to release endothelium-derived contracting factors (EDCF). The possible players of endothelium-dependent contractions and the underlying mechanisms leading to the release of EDCF will be discussed in the present review. EDCF is likely to consist of two components: 1) prostanoids (including endoperoxides, prostacyclin, thromboxane A(2), and prostaglandin E(2)) and 2) reactive oxygen species. The former directly activate thromboxane/prostaglandin endoperoxide (TP) receptors of the vascular smooth muscle cells which leads to their contraction, while the latter first stimulate the cyclooxygenase in the smooth muscle with subsequent stimulation of the TP receptors by the prostanoids produced. Dysfunction in calcium handling is the leading causal factor for the exacerbated occurrence of endothelium-dependent contractions in the aorta of the spontaneously hypertensive rat (SHR). The observed increased expressions of endothelial COX-1, prostacyclin synthase, thromboxane synthase and enhanced TP receptor sensitivity are not prerequisites for, but intensify the magnitude of endothelium-dependent contractions. Selective TP receptor antagonists are effective in preventing endothelium-dependent contractions in vitro which highlights the prospective use of such drugs in correcting the imbalanced release of endothelium-derived vasoactive substances that accompany vascular disease. PMID- 19285527 TI - Quantitative effects of position and type of single mismatch on single base primer extension. AB - Single mismatch (MM) present at the region where primer binds onto the template strand can greatly affect the PCR efficacy. Earlier studies revealed that PCR or primer extension is hindered by a single MM at the primer 3' end. The MMs located at other positions within a primer also have similar performance, but to what extent they can decrease the efficiency is not clear. In this study, a modified single base extension assay was used to systematically compare the extension efficiencies between a perfect-matched (PM) primer and its single-MM primers with all possible MM types. The extension efficiencies of single-MM primers, which were generally lower or equivalent to that of the PM primer, were observed to strongly depend on the MM location and/or type. Due to the enzymatic activity, single MMs present at the last 3-4 positions from the primer 3' end exhibited zero or minimal (<3.9%) extension efficiencies. For those MMs at positions 5 onward from primer 3' end where was affected mainly by the primer-target binding stability, an increasing trend in extension efficiency with the highest (i.e., 69.3%) occurring at the primer 5' end was observed to significantly correlate in an inverse relationship with the duplex stability (i.e., difference of melting temperature) under a empirically polynomial equation, y=-0.0731 x(3) + 2.2519 x(2) - 22.617 x + 76.691 (R(2)=0.5318). It was further shown that the extension efficiencies of these MM types could be improved with a factor of 3.25 on average in relation to the decrease in the annealing temperature by 7 degrees C. On the other hand, substitution of a less selective inosine nucleotide did not convincingly improve the extension efficiency. Overall findings obtained could further improve the rational design of oligonucleotide primers in various microbiological studies that involve the use of PCR techniques. PMID- 19285528 TI - Oxaliplatin encapsulated in PEG-coated cationic liposomes induces significant tumor growth suppression via a dual-targeting approach in a murine solid tumor model. AB - We recently designed a PEG-coated cationic liposome targeted to angiogenic vessels and showed, in a murine dorsal air sac model, potent anti-angiogenic activity of an oxaliplatin (l-OHP) formulation of this liposome. In the present study, we extended the l-OHP formulation to a murine tumor-xenograft model. Following three injections, l-OHP containing PEG-coated cationic liposomes showed substantial tumor growth suppression and increased survival time of tumor-bearing mice without apparent side effects, compared with other l-OHP containing PEG coated neutral liposomes and free l-OHP. In vivo imaging showed a preferential tumor accumulation and a broader distribution of PEG-coated cationic liposomes, compared with PEG-coated neutral liposomes. In addition, PEG-coated cationic liposomes delivered larger amounts of l-OHP into the tumor tissue than other l OHP formulations, correlating with its antitumor efficiency. In vitro studies indicated that PEG-coated cationic liposomes were internalized not only by tumor cells but also by endothelial cells, and consequently its l-OHP formulation displayed higher cytotoxicity towards both cell types as compared with l-OHP containing PEG-coated neutral liposomes. In summary, l-OHP containing PEG-coated cationic liposomes induced significant tumor growth suppression, presumably by delivering encapsulated l-OHP into both tumor endothelial cells and tumor cells. Such dual targeting approach, i.e. vascular-targeting and tumor-targeting with a single liposomal l-OHP formulation, may have great potential for overcoming some major limitations in conventional chemotherapy. PMID- 19285529 TI - A novel method for monitoring the cytosolic delivery of peptide cargo. AB - The intracellular delivery of a diverse array of cargos can be mediated by conjugation to cell-penetrating peptides (CPPs). To date, delivery of cargos into the cytosol via CPPs has been measured indirectly and normally, has been inferred from changes in biological activity. We describe a novel method to directly assay CPP-mediated delivery of peptide cargo into the cytosol, and use this method to define the kinetics of this process. The CPP and the cargo are differentially labeled with the fluorophores FAM (carboxyfluorescein), and TAMRA (carboxytetramethylrhodamine) respectively, and coupled via a disulfide bond to promote quenching of FAM fluorescence by the proximal TAMRA. Delivery of the peptide pair to cells produces an increase in FAM fluorescence within 10 min, consistent with its rapid transfer into the reducing environment of the cytosol, separation of the two components, and concomitant dequenching. The fluorescence based assay described here can thus be used to select a CPP module that is optimized for efficient delivery of particular cargos designed to modify molecular targets in the cytosol. PMID- 19285530 TI - Controlled release of insulin from pH/temperature-sensitive injectable pentablock copolymer hydrogel. AB - A pH- and temperature-sensitive hydrogel of poly(beta-amino ester)-poly(epsilon caprolactone)-poly (ethylene glycol)-poly(epsilon-caprolactone)-poly(beta-amino ester) (PAE-PCL-PEG-PCL-PAE) pentablock copolymer was evaluated as a sustained injectable insulin delivery system. Insulin was readily loaded into the matrix, forming an ionically linked insulin-PAE complex. Complex mixtures containing various concentrations of insulin and copolymer were subcutaneously injected into male Sprague-Dawley rats to study the profile of insulin release in vivo. The insulin-release profile showed that insulin was maintained at a constant steady state level for 15 days, and further demonstrated that insulin levels were controlled by the amount of insulin loaded into the copolymer and the copolymer concentration in the hydrogel. The effect of the insulin-gel complex was further investigated in the streptozotocin (STZ) diabetic rat model. After subcutaneously injecting complex mixtures into STZ-induced diabetic rats, blood glucose and plasma insulin levels were measured. The results showed that the diabetic rats could be treated for more than 1 week with a single injection of the complex mixture containing 10 mg/mL insulin in a 30 wt.% copolymer solution, suggesting that this pH/temperature-sensitive insulin-hydrogel complex system may have therapeutic potential. PMID- 19285531 TI - Low cholesterol, impulsivity and the economy. PMID- 19285533 TI - Effects of green tea catechin on embryo/fetal development in rats. AB - Evidence suggests that the health benefits associated with green tea consumption are related to tea catechins. The objective of this study was to evaluate potential maternal and fetal effects of standardized heat-sterilized green tea catechins (GTC-H). GTC-H was gavage administered to mated female rats from gestation day 6 through 17, at doses of 0, 200, 600, and 2000 mg/kg/day. There were no GTC-H-related deaths or macroscopic findings. During the entire gestation period in the high-dose (2000 mg/kg/day)-treated group and during days 6-9 and 6 18 in the 600 mg/kg/day group, mean body weight gain was lower. Mean feed consumption was lower during gestation days 6-9 in the 600 mg/kg/day group and during gestation days 6-9 and 9-12 in the 2000 mg/kg/day group. Compared to the control group, mean body weights in the 600 and 2000 mg/kg/day groups were up to 5.1% and 7.7% lower during gestation days 9-20. GTC-H administration did not affect mean gravid uterine weights or intrauterine growth and survival. There were no GTC-H-related fetal malformations or developmental variations. Based on the results of this study, the no-observed-adverse-effect level (NOAEL) for GTC-H was 200mg/kg/day for maternal toxicity, and 2000 mg/kg/day for embryo/fetal development. PMID- 19285534 TI - Acetylcholinesterase inhibitory and antioxidant properties of Cyclotrichium niveum, Thymus praecox subsp. caucasicus var. caucasicus, Echinacea purpurea and E. pallida. AB - The dichloromethane, ethyl acetate, ethanol, and aqueous extracts of Cyclotrichium niveum (CN) and Thymus praecox subsp. caucasicus var. caucasicus (TP), Echinacea purpurea (EPU), and E. pallida (EPA) along with the essential oils of CN and TP were assessed for their anti-acetylcholinesterase (AChE) and antioxidant activities. AChE inhibition was estimated using spectrophotometric method of Ellman. Antioxidant activity was evaluated by 2,2-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging and ferrous ion-chelating power tests. Ferric-reducing antioxidant power (FRAP) of CN and TP were also tested. CN essential oil was found to contain isomenthone (56.21%) and pulegone (19.76%). The ethyl acetate (83.11-87.98%) and dichloromethane (73.45-84.02%) extracts of CN showed the highest AChE inhibition. The ethyl acetate and ethanol extracts of TP exerted significant DPPH scavenger effect. The water extracts of CN and TP and the chloroform extract of the aerial parts of EPU displayed the highest ferrous ion-chelating effect. The leaf and flower essential oils of TP had the best FRAP. PMID- 19285535 TI - Prophylactic role of curcumin in dextran sulfate sodium (DSS)-induced ulcerative colitis murine model. AB - We have addressed in this study the possible protective role of the main principle of turmeric pigment; curcumin on a murine model of ulcerative colitis (UC). Colitis was induced by administration of dextran sulfate sodium (DSS) (3% W/V) in drinking water to male Swiss albino rats for 5 consecutive days. DSS challenge induced UC model that was well characterized morphologically and biochemically. DSS produced shrinkage of colon length and increased the relative colon weight/length ratio accompanied by mucosal edema and bloody stool. Histologically, DSS produced submucosal erosions, ulceration, inflammatory cell infiltration and crypt abscess as well as epithelioglandular hyperplasia. The model was confirmed biochemically, and the test battery entailed elevated serum tumor necrosis factor (TNF-alpha) and colonic activity of myleoperoxidase (MPO). Colonic glutathione-S-transferase (GST) activity and its substrate concentration; GSH, were notably reduced, while lipid peroxidation, expressed as malondialdehyde (MDA) level, and total nitric oxide (NO) were significantly increased. Prior administration of curcumin (100mg/kg, IP) for 7 consecutive days ahead of DSS challenge mitigated the injurious effects of DSS and ameliorated all the altered biochemical parameters. These results suggest that curcumin could possibly have a protective role in ulcerative colitis probably via regulation of oxidant/anti oxidant balance and modulation of the release of some inflammatory endocoids, namely TNF-alpha and NO. PMID- 19285536 TI - Effects of borneol on the level of DNA damage induced in primary rat hepatocytes and testicular cells by hydrogen peroxide. AB - The aim of this paper was to evaluate genotoxic effects of borneol and its ability to change DNA-damaging effects of H2O2 in rat hepatocytes and testicular cells. Both in vitro and ex vivo approaches were used in the case of hepatocytes. Testicular cells were tested only ex vivo, i.e. shortly after isolation from rats supplemented by borneol. Cytotoxicity of borneol increased in in vitro conditions in a concentration-dependent manner and it was associated with DNA-damaging effects at toxic concentrations. While non-toxic concentrations of borneol applied in vitro protected cells against H2O2-induced DNA damage and interfered only partly with rejoining of H2O2-induced DNA strand breaks, cytotoxic concentrations of borneol manifested synergy with H2O2, i.e. enhanced DNA damaging effects of H2O2. On the other side, borneol given to rats in drinking water decreased the level of DNA damage induced by H2O2 in both hepatocytes and testicular cells. Our results show that though at higher concentrations (2-h treatment with >2 mM borneol >0.3084 mg/ml) borneol acts cytotoxically and genotoxically on primary hepatocytes cultured in vitro, if given to rats during 7 days in a daily concentration of 17.14 or 34.28 mg/kg it reduces genotoxicity of H2O2 in both hepatocytes and testicular cells. PMID- 19285537 TI - Various strategies for obtaining artificial hemoproteins: from "hemoabzymes" to "hemozymes". AB - The design of artificial hemoproteins that could lead to new biocatalysts for selective oxidation reactions of organic compounds presents a huge interest especially in pharmacology, both for a better understanding of the metabolic profile of drugs and for the synthesis of enantiomerically pure molecules that could be involved in the design of drugs. The present results show that the so called "host-guest strategy" that involves the non-covalent incorporation of anionic water-soluble iron-porphyrins into xylanase A from Streptomyces lividans, a low cost protein, leads to such an artificial hemoprotein that is able to perform the stereoselective oxidation of sulfides. PMID- 19285538 TI - Coordination of bismuth and lead in porphyrins: towards an in-situ generator for alpha-radiotherapy? AB - In order to investigate the possible coordination of both bismuth(III) and lead(II) by porphyrins for an application in alpha-radioimmunotherapy, several series of functionalized bismuth porphyrins have been synthesized and their stabilities in acidic medium were compared. Where unfunctionalized porphyrins are not suitable for such a purpose, strapped hanging carboxylate porphyrins are stable and allow a rapid coordination of both bismuth and lead. This property could lead to the preparation of an in-situ generator of alpha particles by a pre complexation of (212)lead, the radioactive parent of (212)bismuth, in the ligand. PMID- 19285539 TI - The selenium to selenoprotein pathway in eukaryotes: more molecular partners than anticipated. AB - The amino acid selenocysteine (Sec) is the major biological form of the trace element selenium. Sec is co-translationally incorporated in selenoproteins. There are 25 selenoprotein genes in humans, and Sec was found in the active site of those that have been attributed a function. This review will discuss how selenocysteine is synthesized and incorporated into selenoproteins in eukaryotes. Sec biosynthesis from serine on the tRNA(Sec) requires four enzymes. Incorporation of Sec in response to an in-frame UGA codon, otherwise signaling termination of translation, is achieved by a complex recoding machinery to inform the ribosomes not to stop at this position on the mRNA. A number of the molecular partners acting in this machinery have been identified but their detailed mechanism of action has not been deciphered yet. Here we provide an overview of the literature in the field. Particularly striking is the higher than originally envisaged number of factors necessary to synthesize Sec and selenoproteins. Clearly, selenoprotein synthesis is an exciting and very active field of research. PMID- 19285541 TI - Combustion smoke exposure induces up-regulated expression of vascular endothelial growth factor, aquaporin 4, nitric oxide synthases and vascular permeability in the retina of adult rats. AB - Retinal cells respond to various experimental stimuli including hypoxia, yet it remains to be investigated whether they react to smoke inhalation. We show here that retinal cells in rats, notably the ganglion cells, Muller cells, astrocytes and blood vessels responded vigorously to a smoke challenge. The major changes included up-regulated expression of vascular endothelial growth factor (VEGF), aquaporin 4 (AQP4) and nitric oxide synthase (NOS). VEGF expression was localized in the ganglion cells, Muller cells, astrocytes and associated blood vessels. AQP4 was markedly enhanced in both astrocytes and Muller cells. Increase in vascular permeability after smoke exposure was evidenced by extravasation of serum derived rhodamine isothiocyanate which was internalized by Muller cells and ganglion cells. The tracer leakage was attenuated by aminoguanidine and N(G) nitro-L-arginine methyl ester (L-NAME) treatment which suppressed retinal tissue NOS and nitric oxide (NO) levels concomitantly. It is suggested that VEGF, AQP4 and NO are involved in increased vascular permeability following acute smoke exposure in which hypoxia was ultimately implicated as shown by blood gases analysis. NOS inhibitors effectively reduced the vascular leakage and hence may ameliorate possible retinal edema in smoke inhalation. PMID- 19285542 TI - Single administration of 1-benzyl-1,2,3,4-tetrahydroisoquinoline increases the extracellular concentration of dopamine in rat striatum. AB - We performed a combined neurochemical and behavioral study to determine the effects of 1-benzyl-1,2,3,4-tetrahydroisoquinoline (1-BnTIQ) on the extracellular dopamine concentrations in the striatum. Single dose administration of 1-BnTIQ (20, 40, and 80 mg/kg i.p.) increased striatal dopamine extracellular levels in a dose-dependent manner when an in vivo microdialysis technique was used to assess dopamine levels in the striatum of rats. Enhancement of striatal dopamine levels by systemic administration of a single dose of 1-BnTIQ was suppressed by perfusion of tetrodotoxin and a calcium ion-free solution into the striatum. This 1-BnTIQ-induced increase in extracellular dopamine concentration was also inhibited by pre-treatment with a dopamine uptake inhibitor, GBR12909 (1-(2 [bis(4-Fluorophenyl)-4-(3-phenylpropyl)piperazine dihydrochloride). Local application of 1-BnTIQ into the striatum via a dialysis probe failed to enhance the extracellular concentration of dopamine. However, microinjection of 1-BnTIQ into the substantia nigra pars compacta increased the extracellular dopamine levels in the striatum. Locomotor activity was increased by systemic administration of a single dose of 1-BnTIQ in a dose-dependent manner. This 1 BnTIQ-induced locomotor activity was attenuated by pre-treatment with SCH23390 (R(+)-7-Chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochlodride) and raclopride, D(1) and D(2) dopaminergic receptor antagonists, respectively. Moreover, 1-BnTIQ induced ipsilateral rotational behavior in 6 hydroxydopamine-lesioned rats. These results suggest that systemic administration of a single dose of 1-BnTIQ increases striatal extracellular dopamine concentration through activation of dopaminergic nigra striatal neurons via the dopamine transporter. PMID- 19285543 TI - Visual motion direction is represented in population-level neural response as measured by magnetoencephalography. AB - We investigated whether direction information is represented in the population level neural response evoked by the visual motion stimulus, as measured by magnetoencephalography. Coherent motions with varied speed, varied direction, and different coherence level were presented using random dot kinematography. Peak latency of responses to motion onset was inversely related to speed in all directions, as previously reported, but no significant effect of direction on latency changes was identified. Mutual information entropy (IE) calculated using four-direction response data increased significantly (>2.14) after motion onset in 41.3% of response data and maximum IE was distributed at approximately 20 ms after peak response latency. When response waveforms showing significant differences (by multivariate discriminant analysis) in distribution of the three waveform parameters (peak amplitude, peak latency, and 75% waveform width) with stimulus directions were analyzed, 87 waveform stimulus directions (80.6%) were correctly estimated using these parameters. Correct estimation rate was unaffected by stimulus speed, but was affected by coherence level, even though both speed and coherence affected response amplitude similarly. Our results indicate that speed and direction of stimulus motion are represented in the distinct properties of a response waveform, suggesting that the human brain processes speed and direction separately, at least in part. PMID- 19285540 TI - Cancer gene discovery in mouse and man. AB - The elucidation of the human and mouse genome sequence and developments in high throughput genome analysis, and in computational tools, have made it possible to profile entire cancer genomes. In parallel with these advances mouse models of cancer have evolved into a powerful tool for cancer gene discovery. Here we discuss the approaches that may be used for cancer gene identification in both human and mouse and discuss how a cross-species 'oncogenomics' approach to cancer gene discovery represents a powerful strategy for finding genes that drive tumourigenesis. PMID- 19285545 TI - Characterization and mapping of sleep-waking specific neurons in the basal forebrain and preoptic hypothalamus in mice. AB - We recorded 872 single units across the complete sleep-waking cycle in the mouse preoptic area (POA) and basal forebrain (BFB), which are deeply involved in the regulation of sleep and wakefulness (W). Of these, 552 were sleep-active, 96 were waking-active, 106 were active during both waking and paradoxical sleep (PS), and the remaining 118 were state-indifferent. Among the 872, we distinguished slow wave sleep (SWS)-specific, SWS/PS-specific, PS-specific, W-specific, and W/PS specific neurons, the last group being further divided into specific tonic type I slow (TI-Ss) and specific tonic type I rapid (TI-Rs) both discharging specifically in association with cortical activation during both W and PS. Both the SWS/PS-specific and PS-specific neurons were distributed throughout a wide region of the POA and BFB, whereas the SWS-specific neurons were mainly located in the middle and ventral half of the POA and adjacent BFB, as were the W specific and W/PS-specific neurons. At the transition from waking to sleep, the majority of SWS-specific and all SWS/PS-specific neurons fired after the onset of cortical synchronization (deactivation), whereas all W-specific and W/PS-specific neurons showed a significant decrease in firing rate >0.5 s before the onset. At the transition from SWS to W, the sleep-specific neurons showed a significant decrease in firing rate 0.1 s before the onset of cortical activation, while the W-specific and W/PS-specific neurons fired >0.5 s before the onset. TI-Ss neurons were characterized by a triphasic broad action potential, slow single isolated firing, and an antidromic response to cortical stimulation, whereas TI-Rs neurons were characterized by a narrow action potential and high frequency burst discharge in association with theta waves in PS. These data suggest that the forebrain sleep/waking switch is regulated by opposing activities of sleep promoting (SWS-specific and SWS/PS-specific) and waking-promoting (W-specific and W/PS-specific) neurons, that the initiation of sleep is caused by decreased activity of the waking-promoting neurons (disfacilitation), and that the W/PS specific neurons are deeply involved in the processes of cortical activation/deactivation. PMID- 19285546 TI - Ventrolateral prefrontal cortex activity associated with individual differences in arbitrary delayed paired-association learning performance: a functional magnetic resonance imaging study. AB - To describe the neural substrates of successful episodic long-term memory encoding, we collected functional magnetic-resonance imaging data as participants completed an arbitrary delayed auditory paired-association learning task. During the task, subjects learned predefined but hidden stimulus pairs by trial and error based on visual feedback. Delay period activity represents the retrieval of the relationship between the cue item and its candidate for associates, that is, working memory. Our hypothesis was that the neural substrates of working memory would be related to long-term memory encoding in a performance-dependent manner. Thus, inter-individual variance in performance following a fixed learning set would be associated with differing neural activations during the delay period. The number of learning trials was adjusted such that performance following completion of the learning set varied across subjects. Each trial consisted of the successive presentation of two stimuli (first stimulus and second stimulus [S2]) with a fixed delay interval, allowing extraction of sustained activity during the delay period. Sustained activities during the delay period were found in the bilateral dorsolateral prefrontal cortex, intraparietal sulcus, and left ventrolateral prefrontal cortex, as well as the premotor and pre-supplementary motor areas. The activities did not change in strength across learning, suggesting that these effects represent working memory components. The sustained activity in the ventrolateral prefrontal region was correlated with task performance. Task performance was also positively correlated with the decrement in S2/feedback-related activity during learning in the superior temporal sulcus, a region previously shown to be involved in association learning. These findings are consistent with lesion and neuroimaging studies showing that the ventrolateral prefrontal cortex plays an important role in long-term memory encoding, and raise the possibility that working memory processes interact with long-term memory formation as represented by the covariation of activity in the superior temporal sulcus and the ventrolateral prefrontal cortex. PMID- 19285547 TI - Extracellular divalent cations modulate aminoglycoside-induced hair cell death in the zebrafish lateral line. AB - Aminoglycoside antibiotics cause death of sensory hair cells. Research over the past decade has identified several key players in the intracellular cascade. However, the role of the extracellular environment in aminoglycoside ototoxicity has received comparatively little attention. The present study uses the zebrafish lateral line to demonstrate that extracellular calcium and magnesium ions modulate hair cell death from neomycin and gentamicin in vivo, with high levels of either divalent cation providing significant protection. Imaging experiments with fluorescently-tagged gentamicin show that drug uptake is reduced under high calcium conditions. Treating fish with the hair cell transduction blocker amiloride also reduces aminoglycoside uptake, preventing the toxicity, and experiments with variable calcium and amiloride concentrations suggest complementary effects between the two protectants. Elevated magnesium, in contrast, does not appear to significantly attenuate drug uptake, suggesting that the two divalent cations may protect hair cells from aminoglycoside damage through different mechanisms. These results provide additional evidence for calcium- and transduction-dependent aminoglycoside uptake. Divalent cations provided differential protection from neomycin and gentamicin, with high cation concentrations almost completely protecting hair cells from neomycin and acute gentamicin toxicity, but offering reduced protection from continuous (6 h) gentamicin exposure. These experiments lend further support to the hypothesis that aminoglycoside toxicity occurs via multiple pathways in a both a drug and time course-specific manner. PMID- 19285548 TI - The absence of a mitochondrial genome in rho0 yeast cells extends lifespan independently of retrograde regulation. AB - The absence of mtDNA in rho0 yeast cells affects both respiration and mitochondrial-nuclear communication (e.g., retrograde regulation, intergenomic signaling, or pleiotropic drug resistance). Previously, it has been reported that some rho0 strains have increased replicative lifespans, attributable to the lack of respiration and retrograde regulation. Here, we have been able to confirm that rho0 cells exhibit increased replicative lifespans but have found that this is not associated with the lack of respiration or reduced oxidative stress but instead, is related to the lack of mtDNA per se in rho0 cells. Also, we find no correlation between the strength of retrograde regulation and lifespan. Furthermore, we find that pdr3- or rtg2- mutations are not responsible for lifespan extension in rho0 cells, ruling out a specific role for PDR3-pleiotropic drug resistance or RGT2-retrograde regulation pathways in the extended lifespans of rho0 cells. Surprisingly, Rtg3p, which acts downstream of Rtg2p, is required for lifespan increase in rho0 cells. Together, these findings indicate that the loss of mtDNA per se and not the lack of respiration lead to extended longevity in rho0 cells. They also suggest that Rtg3p, acting independently of retrograde regulation, mediates this effect, possibly via intergenomic signaling. PMID- 19285549 TI - Genotoxicity of the neurotransmitter dopamine in vitro. AB - Alterations in dopamine levels play a role in several human pathological conditions and their pharmacological treatment. Here we describe an induction of genomic damage detected as micronucleus formation by concentrations in the low micromolar range (6.25-25 microM) in three cell lines in vitro. Rat neuronal PC12 cells exhibited a more pronounced induction (about 10-fold over control at 100 microM) than human lymphoblastoid TK6 cells and rat kidney NRK cells (about 2 fold over control at 100 microM). The role of transporters and receptors in the formation of genomic damage was investigated in PC12 cells, in which the effect of dopamine was reduced by addition of the antioxidants TEMPOL and dimethylthiourea, by inhibitors of the dopamine transporter (GBR 12909 and nomifensine) and by a D2 antagonist (sulpiride). Antioxidative effects of nomifensine and sulpiride, but not of GBR 12909, were excluded, since they did not protect oxidative stress sensitive HL-60 cells from hydrogen peroxide-induced damage in the comet assay. Thus, the transport of dopamine into the cell and the signalling upon binding to D2 receptors was required for the genotoxic effect of dopamine in PC12 cells, which was mediated by intracellular dopamine oxidation products and/or reactive oxygen species. PMID- 19285550 TI - Involvement of hydrogen peroxide in histamine-induced modulation of WM35 human malignant melanoma cell proliferation. AB - Histamine is a recognized growth factor in melanoma, and exogenous histamine produces a dual effect on proliferation. We have previously reported that histamine at micromolar concentrations reduces the proliferation of melanoma cell lines. To investigate the mechanism by which histamine inhibits proliferation of WM35 human melanoma cells, we have studied the involvement of histamine in reactive oxygen species production and antioxidant enzyme regulation in these cells. Results indicate that histamine treatment (10 microM) significantly increased hydrogen peroxide levels, whereas it slightly decreased superoxide levels associated with an enhancement of superoxide dismutase and a reduction in catalase activity. Additionally, catalase treatment reversed the inhibitory effect of histamine on proliferation, and various treatments that reduce hydrogen peroxide formation increased proliferation of these cells. Furthermore, we demonstrate that the inhibition of proliferation produced by histamine was mediated at least in part by an induction of cell senescence. We conclude that hydrogen peroxide is involved in histamine-mediated modulation of proliferation in malignant melanoma cells. PMID- 19285552 TI - Dual positive and negative regulation of GPCR signaling by GTP hydrolysis. AB - G protein-coupled receptors (GPCRs) regulate a variety of intracellular pathways through their ability to promote the binding of GTP to heterotrimeric G proteins. Regulator of G protein signaling (RGS) proteins increases the intrinsic GTPase activity of Galpha-subunits and are widely regarded as negative regulators of G protein signaling. Using yeast we demonstrate that GTP hydrolysis is not only required for desensitization, but is essential for achieving a high maximal (saturated level) response. Thus RGS-mediated GTP hydrolysis acts as both a negative (low stimulation) and positive (high stimulation) regulator of signaling. To account for this we generated a new kinetic model of the G protein cycle where Galpha(GTP) enters an inactive GTP-bound state following effector activation. Furthermore, in vivo and in silico experimentation demonstrates that maximum signaling output first increases and then decreases with RGS concentration. This unimodal, non-monotone dependence on RGS concentration is novel. Analysis of the kinetic model has revealed a dynamic network motif that shows precisely how inclusion of the inactive GTP-bound state for the Galpha produces this unimodal relationship. PMID- 19285551 TI - Cytochrome c/cardiolipin relations in mitochondria: a kiss of death. AB - Recently, phospholipid peroxidation products gained a reputation as key regulatory molecules and participants in oxidative signaling pathways. During apoptosis, a mitochondria-specific phospholipid, cardiolipin (CL), interacts with cytochrome c (cyt c) to form a peroxidase complex that catalyzes CL oxidation; this process plays a pivotal role in the mitochondrial stage of the execution of the cell death program. This review is focused on redox mechanisms and essential structural features of cyt c's conversion into a CL-specific peroxidase that represent an interesting and maybe still unique example of a functionally significant ligand change in hemoproteins. Furthermore, specific characteristics of CL in mitochondria--its asymmetric transmembrane distribution and mechanisms of collapse, the regulation of its synthesis, remodeling, and fatty acid composition--are given significant consideration. Finally, new concepts in drug discovery based on the design of mitochondria-targeted inhibitors of cyt c/CL peroxidase and CL peroxidation with antiapoptotic effects are presented. PMID- 19285553 TI - Microarray analyses of the effects of NF-kappaB or PI3K pathway inhibitors on the LPS-induced gene expression profile in RAW264.7 cells: synergistic effects of rapamycin on LPS-induced MMP9-overexpression. AB - Lipopolysaccharide (LPS) activates a broad range of signalling pathways including mainly NF-kappaB and the MAPK cascade, but recent evidence suggests that LPS stimulation also activates the PI3K pathway. To unravel the specific roles of both pathways in LPS signalling and gene expression profiling, we investigated the effects of different inhibitors of NF-kappaB (BAY 11-7082), PI3K (wortmannin and LY294002) but also of mTOR (rapamycin), a kinase acting downstream of PI3K/Akt, in LPS-stimulated RAW264.7 macrophages, analyzing their effects on the LPS-induced gene expression profile using a low density DNA microarray designed to monitor the expression of pro-inflammatory genes. After statistical and hierarchical cluster analyses, we determined five clusters of genes differentially affected by the four inhibitors used. In the fifth cluster corresponding to genes upregulated by LPS and mainly affected by BAY 11-7082, the gene encoding MMP9 displayed a particular expression profile, since rapamycin drastically enhanced the LPS-induced upregulation at both the mRNA and protein levels. Rapamycin also enhanced the LPS-induced NF-kappaB transactivation as determined by a reporter assay, phosphorylation of the p38 and Erk1/2 MAPKs, and counteracted PPAR activity. These results suggest that mTOR could negatively regulate the effects of LPS on the NF-kappaB and MAPK pathways. We also performed real-time RT-PCR assays on mmp9 expression using rosiglitazone (agonist of PPARgamma), PD98059 (inhibitor of Erk 1/2) and SB203580 (inhibitor of p38(MAPK)), that were able to counteract the rapamycin mediated overexpression of mmp9 in response to LPS. Our results suggest a new pathway involving mTOR for regulating specifically mmp9 in LPS-stimulated RAW264.7 cells. PMID- 19285554 TI - Deficiency of polycystic kidney disease-1 gene (PKD1) expression increases A(3) adenosine receptors in human renal cells: implications for cAMP-dependent signalling and proliferation of PKD1-mutated cystic cells. AB - Cyst growth and expansion in autosomal dominant polycystic kidney disease (ADPKD) has been attributed to numerous factors, including ATP, cAMP and adenosine signalling. Although the role of ATP and cAMP has been widely investigated in PKD1-deficient cells, no information is currently available on adenosine-mediated signalling. Here we investigate for the first time the impact of abnormalities of polycystin-1 (PC1) on the expression and functional activity of adenosine receptors, members of the G-protein-coupled receptor superfamily. Pharmacological, molecular and biochemical findings show that a siRNA-dependent PC1-depletion in HEK293 cells and a PKD1-nonsense mutation in cyst-derived cell lines result in increased expression of the A(3) adenosine receptor via an NFkB dependent mechanism. Interestingly, A(3) adenosine receptor levels result higher in ADPKD than in normal renal tissues. Furthermore, the stimulation of this receptor subtype with the selective agonist Cl-IB-MECA causes a reduction in both cytosolic cAMP and cell proliferation in both PC1-deficient HEK293 cells and cystic cells. This reduction is associated with increased expression of p21(waf) and reduced activation not only of ERK1/2, but also of S6 kinase, the main target of mTOR signalling. In the light of these findings, the ability of Cl-IB-MECA to reduce disease progression in ADPKD should be further investigated. Moreover, our results suggest that NFkB, which is markedly activated in PC1-deficient and cystic cells, plays an important role in modulating A(3)AR expression in cystic cells. PMID- 19285555 TI - Revisiting the mystery of fibronectin multimers: the fibronectin matrix is composed of fibronectin dimers cross-linked by non-covalent bonds. AB - Fibronectin (FN) matrix fibrils have long been thought to be formed by disulfide bonded FN multimers, although there is no direct evidence that they are covalently linked with each other. To understand the biochemical properties of these fibrils, we extracted a crude FN matrix from FN-YPet transfected 3T3 cell culture using 0.2% deoxycholate and DNase. The insoluble extracted matrix preserved fibrillar structures and a major portion of the extracted proteins migrated as FN monomers on an SDS gel under reducing conditions. Under non reducing conditions, some FN molecules appeared to be trapped at the top of the stacking gel. We tested this by mixing fluorescently labeled FN dimers with the extracted matrix just before loading on an SDS gel, and found that most of them were trapped with the extracted proteins at the top of the stacking gel. These results suggested that some components of the extracted matrix plugged the stacking gel and FN dimers were trapped with them. Rotary shadowing electron microscopy showed that the extracted matrix had some fibers that resembled fibrillin microfibrils. Peptide mass fingerprinting confirmed the presence of fibrillin in the extracted matrix. Fibrillin is known to form disulfide-bonded multimers and it is likely to be one of the components that plug the stacking gel and trap FN molecules in this system. The phenomenon by which FN molecules appear to migrate as multimers on SDS gels is thus an artifact rising from the presence of other large components in the extract. We conclude that FN matrix fibrils are made of FN dimers that are further cross-linked by non-covalent protein-protein bonds. PMID- 19285556 TI - Scintillation proximity assays for mechanistic and pharmacological analyses of HIV-1 integration. AB - The early events of HIV-1 replication are highlighted by reverse transcription and integration, and the reverse transcriptase and integrase enzymes are important therapeutic targets. Integration proceeds through a series of steps including assembly of integrase on the viral donor DNA ends, 3'-processing, and DNA strand transfer. First generation integrase assays typically included all biochemical reagents in solution where excess donor substrate could serve as the target for DNA strand transfer. These conditions, though valuable for understanding mechanistic aspects of HIV-1 integration, fell short of critical pharmacological designs as most early inhibitors were found to block assembly instead of enzyme function. Second generation designs, which decoupled assembly from DNA strand transfer, afforded the specificity required to identify clinically relevant compounds. Here, we describe versatile scintillation proximity-based assays whereby integrase is assembled onto donor DNA that is immobilized onto the surface of beads. Immobilization and subsequent washing of excess donor DNA eliminates its potential to serve as target DNA, allowing investigation of the DNA strand transfer reaction in isolation. Assembled complexes can be used in high-throughput DNA strand transfer assays if radio labeled target DNA is employed or in integrase binding assays using a suitable radioligand. PMID- 19285557 TI - Ultrastructural detection of nucleic acids within heat shock-induced perichromatin granules of HeLa cells by cytochemical and immunocytological methods. AB - The perichromatin granules (PGs) are enigmatic structures of the cell nucleus. The major drawbacks for a biological study are their rare occurrence and their small size in normal conditions. As heat shock has been shown to increase their number, we applied a hyperthermal shock on HeLa cells to investigate the nucleic acid content of PGs by means of cytochemical and immunocytological approaches. These heat shock-induced PGs (hsiPGs) appeared as clusters organized in the form of honeycomb structures and were always associated with some blocks of condensed chromatin, such as the perinucleolar chromatin shell. A stalk connecting the hsiPG to the chromatin could be observed. For the detection of RNA, we applied an immunocytological method involving two anti-RNA antibodies and quantified the gold labelling obtained. The results clearly revealed that hsiPGs contained RNA. Regarding to the detection of DNA, we used three different methods followed by quantitative analyses. The results seemed to indicate that a small amount of DNA was present in hsiPGs. Together, these findings suggest that hsiPGs might be RNP structures associated with particular regions of DNA. PMID- 19285558 TI - Reduced prefrontal cortical gray matter volume in young adults exposed to harsh corporal punishment. AB - OBJECTIVE: Harsh corporal punishment (HCP) during childhood is a chronic, developmental stressor associated with depression, aggression and addictive behaviors. Exposure to traumatic stressors, such as sexual abuse, is associated with alteration in brain structure, but nothing is known about the potential neurobiological consequences of HCP. The aim of this study was to investigate whether HCP was associated with discernible alterations in gray matter volume (GMV) using voxel-based morphometry (VBM). METHODS: 1455 young adults (18-25 years) were screened to identify 23 with exposure to HCP (minimum 3 years duration, 12 episodes per year, frequently involving objects) and 22 healthy controls. High-resolution T1-weighted MRI datasets were obtained using Siemens 3 T trio scanner. RESULTS: GMV was reduced by 19.1% in the right medial frontal gyrus (medial prefrontal cortex; MPFC, BA10) (P=0.037, corrected cluster level), by 14.5% in the left medial frontal gyrus (dorsolateral prefrontal cortex; DLPFC, BA9) (P=0.015, uncorrected cluster level) and by 16.9% in the right anterior cingulate gyrus (BA24) (P<0.001, uncorrected cluster level) of HCP subjects. There were significant correlations between GMV in these identified regions and performance IQ on the WAIS-III. CONCLUSIONS: Exposing children to harsh HCP may have detrimental effects on trajectories of brain development. However, it is also conceivable that differences in prefrontal cortical development may increase risk of exposure to HCP. PMID- 19285559 TI - Pattern of hippocampal shape and volume differences in blind subjects. AB - Numerous studies in animals and humans have shown that the hippocampus (HP) is involved in spatial navigation and memory. Blind subjects, in particular, must memorize extensive information to compensate for their lack of immediate updating of spatial information. Increased demands on spatial cognition and memory may be associated with functional and structural HP plasticity. Here we examined local size and shape differences in the HP of blind and sighted individuals. A 3D parametric mesh surface was generated to represent right and left HPs in each individual, based on manual segmentations of 3D volumetric T1-weighted MR images of 22 blind subjects and 28 matched controls. Using a new surface mapping algorithm described in (Shi, Y., Thompson, P.M., de Zubicaray, G.I., Rose, S.E., Tu, Z., Dinov, I., Toga, A.W., Direct mapping of hippocampal surfaces with intrinsic shape context, NeuroImage, Available online May 24, (In Press).), we created an average hippocampal surface for the controls, and computed its normal distance to each individual surface. Statistical maps were created to visualize systematic anatomical differences between groups, and randomization tests were performed to correct for multiple comparisons. In both scaled and unscaled data, the anterior right HP was significantly larger, and the posterior right HP significantly smaller in blind individuals. No significant differences were found for left HP. These differences may reflect adaptive responses to sensory deprivation, and/or increased functional demands on memory systems. They offer a neuroanatomical substrate for future correlations with measures of navigation performance or functional activations related to variations in cognitive strategies. PMID- 19285560 TI - A protocol for use of medetomidine anesthesia in rats for extended studies using task-induced BOLD contrast and resting-state functional connectivity. AB - The alpha-2-adrenoreceptor agonist, medetomidine, which exhibits dose-dependent sedative effects and is gaining acceptance in small-animal functional magnetic resonance imaging (fMRI), has been studied. Rats were examined on the bench using the classic tail-pinch method with three infusion sequences: 100 microg/kg/h, 300 microg/kg/h, or 100 microg/kg/h followed by 300 microg/kg/h. Stepping the infusion rate from 100 to 300 microg/kg/h after 2.5 h resulted in a prolonged period of approximately level sedation that cannot be achieved by a constant infusion of either 100 or 300 microg/kg/h. By stepping the infusion dosage, experiments as long as 6 h are possible. Functional MRI experiments were carried out on rats using a frequency dependent electrical stimulation protocol-namely, forepaw stimulation at 3, 5, 7, and 10 Hz. Each rat was studied for a four-hour period, divided into two equal portions. During the first portion, rats were started at a 100 microg/kg/h constant infusion. During the second portion, four secondary levels of infusion were used: 100, 150, 200, and 300 microg/kg/h. The fMRI response to stimulation frequency was used as an indirect measure of modulation of neuronal activity through pharmacological manipulation. The frequency response to stimulus was attenuated at the lower secondary infusion dosages 100 or 150 microg/kg/h but not at the higher secondary infusion dosages 200 or 300 microg/kg/h. Parallel experiments with the animal at rest were carried out using both electroencephalogram (EEG) and functional connectivity MRI (fcMRI) methods with consistent results. In the secondary infusion period using 300 microg/kg/h, resting-state functional connectivity is enhanced. PMID- 19285561 TI - Fully automated classification of HARDI in vivo data using a support vector machine. AB - The purpose of this study is the classification of high angular resolution diffusion imaging (HARDI) in vivo data using a model-free approach. This is achieved by using a Support Vector Machine (SVM) algorithm taken from the field of supervised statistical learning. Six classes of image components are determined: grey matter, parallel neuronal fibre bundles in white matter, crossing neuronal fibre bundles in white matter, partial volume between white and grey matter, background noise and cerebrospinal fluid. The SVM requires properties derived from the data as input, the so called feature vector, which should be rotation invariant. For our application we derive such a description from the spherical harmonic decomposition of the HARDI signal. With this information the SVM is trained in order to find the function for separating the classes. The SVM is systematically tested with simulated data and then applied to six in vivo data sets. This new approach is data-driven and enables fully automatic HARDI data segmentation without employing a T1 MPRAGE scan and subjective expert intervention. This was demonstrated on five test in vivo data sets giving robust results. The segmentation results could be used as a priori knowledge for increasing the performance of fibre tracking as well as for other clinical and diagnostic applications of diffusion weighted imaging (DWI). PMID- 19285562 TI - Food-aversive classical conditioning increases a persistent sodium current in molluscan withdrawal interneurons in a transcription dependent manner. AB - In this study we examined changes in a persistent sodium current (I(NaP)) after behavioral aversive classical conditioning in the snail Helix pomatia. We trained animals by pairing food with a mild electric shock that triggered the whole-body withdrawal reflex. This aversive training resulted in transcription dependent long-term associative memory. Isolated central nervous system preparations were set up from trained, random control and naive animals and using two-electrode voltage clamp methods, I(NaP) was activated and measured in identified body withdrawal interneurons RPa3 and LPa3. We show here that in preparations from conditioned animals I(NaP) is increased, suggesting that modifications in intrinsic cellular properties contribute to the formation of the memory trace. Blocking RNA synthesis by systemic injection of actinomycin D (0.12microM) suppressed both memory consolidation in intact animals and the learning-induced increase of I(NaP) in withdrawal interneurons, suggesting that aversive classical conditioning affects sodium channel expression at the transcriptional level. PMID- 19285563 TI - Enhanced post-learning memory consolidation is influenced by arousal predisposition and emotion regulation but not by stimulus valence or arousal. AB - Emotionally arousing stimuli are more memorable than neutral ones and arousal induced after learning enhances later retrieval. However, there is as yet little study of how stimulus qualities might interact with induced arousal and how individual differences might influence the modulation of memory. Thus, the present study examined the effect of arousal induced after learning on memory for words that varied in both arousal and valence quality, as well as the influence of three individual differences factors that are known to influence arousal response: emotional suppression, emotional reappraisal, and arousal predisposition. Seventy-six adults (57 female) viewed and rated 60 words that normatively ranged from high to low in arousal and valence. Ten minutes later, they viewed a 3-min comedic or neutral video clip. Arousal induced after learning enhanced 1-week delayed memory, spanning the lengthy task without preference for word type or serial position, contrasting with reports of arousal effects interacting with stimulus qualities. Importantly, being predisposed to arousal led to greater enhancement of long-term memory modulation, while the use of emotional reappraisal, which reduces arousal responding, inhibited the ability of arousal to induce memory enhancement. Thus, individual differences that influence arousal responding can contribute to or interfere with memory modulation. PMID- 19285564 TI - Structural and functional properties of the Trichosporon asahii glucuronoxylomannan. AB - The virulence attributes of Trichosporon asahii are virtually unknown, despite its growing relevance as causative agent of superficial and invasive diseases in humans. Glucuronoxylomannan (GXM) is a well described virulence factor of pathogenic species in the Cryptococcus genus. GXM is also produced by species of the Trichosporon genus, and both polysaccharides share antigenic determinants, but unlike cryptococcal GXM, relatively little work has been done on trichosporal GXMs. In this study, we analyzed structural and functional aspects of GXM produced by T. asahii and compared them to the properties of the cryptococcal polysaccharide. Trichosporal and cryptococcal GXM shared antigenic reactivity, but the former polysaccharide had smaller effective diameter and negative charge. GXM anchoring to the cell wall was perturbed by dimethylsulfoxide and required interactions of chitin-derived oligomers with the polysaccharide. GXM from T. asahii supernatants are incorporated by acapsular mutants of Cryptococcus neoformans, which renders these cells more resistant to phagocytosis by mouse macrophages. In summary, our results establish that despite similarities in cell wall anchoring, antigenic and antiphagocytic properties, trichosporal and cryptococcal GXMs manifest major structural differences that may directly affect polysaccharide assembly at the fungal surface. PMID- 19285565 TI - Activities of gamma-butyrobetaine dioxygenase and concentrations of carnitine in tissues of pigs. AB - In contrast to other species, less is known about carnitine homeostasis in the pig. This study was performed to yield information about the site of carnitine synthesis and carnitine concentrations in various tissues of pigs (Sus scrofa). We found that among several pig tissues, a considerable activity of gamma butyrobetaine dioxygenase (BBD), the last enzyme of carnitine synthesis, exists, like in humans and several other species, only in liver and kidney. Activity of that enzyme in liver and kidney was lower at birth than in the subsequent weeks of life. Highest carnitine concentrations were found in skeletal muscle and heart. Carnitine concentrations in plasma, liver and kidney at birth were higher than in the subsequent weeks of life in spite of the low BBD activity at birth. In conclusion, this study shows that liver and kidney are the major sites of carnitine synthesis and that neonatal pigs do not have an insufficient carnitine status. PMID- 19285566 TI - Thermal, metabolic, hygric and ventilatory physiology of the sandhill dunnart (Sminthopsis psammophila; Marsupialia, Dasyuridae). AB - We present here the first physiological data for the sandhill dunnart (Sminthopsis psammophila), the second largest (35-44 g) sminthopsine dasyurid marsupial, and report torpor for this species. Their thermoneutral body temperature (34.4 degrees C), thermolability below thermoneutrality (0.062 degrees C degrees C(-1)), and mild hyperthermia above thermoneutrality (35.5 degrees C) are typical of small dunnarts, and dasyurids. Basal metabolic rate (0.80 mL O2 g(-1) h(-1)) is as predicted from mass. Sandhill dunnarts generally conform to the Scholander-Irving model of endothermy, although metabolism increases less than expected and extrapolates to a higher than actual body temperature.Wet (0.22 mL O2 g(-1) h(-1) C(-1)) and dry (2.8 J g(-1) h(-1) degrees C(-1)) thermal conductances were as predicted. Thermoneutral evaporative water loss (1.6 mg g(-1) h(-1)) was only 54% of expected, but this is not significantly different, and more likely reflects variability in the marsupial dataset than an adaptation.Relative water economy resembles that of other small marsupials, rodents and birds, with a point of relative economy of 18 degrees C. Respiratory ventilation closely matches metabolic rate, with minute volume increased at low ambient temperatures by increased breathing rate rather than tidal volume; oxygen extraction was constant at about 17%, except during hyperthermia above the thermoneutrality. Torpor conferred significant energetic and hygric benefits. We found no evidence of deviation from allometrically- and phylogenetically-based expectations despite the sandhill dunnart's arid habitat and large (for a dunnart) body mass. PMID- 19285567 TI - The iron trap: iron, malaria and anemia at the mother-child interface. AB - Iron deficiency causes anemia, but prevents malaria for unknown reasons, thus hindering iron supplementation programs for mothers and children. Iron homeostasis is tightly regulated, including at the mother-fetus interface where iron-malaria relationships are complex. Improved iron status assays, and understanding of malaria protection mechanisms, are needed to manage these disorders. PMID- 19285568 TI - Purification and characterization of a cysteine-rich secretory protein from Philodryas patagoniensis snake venom. AB - Cysteine-rich secretory proteins (CRiSPs) are widespread in reptile venoms, but most have functions that remain unknown. In the present study we describe the purification and characterization of a CRiSP (patagonin) from the venom of the rear-fanged snake Philodryas patagoniensis, and demonstrate its biological activity. Patagonin is a single-chain protein, exhibiting a molecular mass of 24,858.6 Da, whose NH(2)-terminal and MS/MS-derived sequences are nearly identical to other snake venom CRiSPs. The purified protein hydrolyzed neither azocasein nor fibrinogen, and it could induce no edema, hemorrhage or inhibition of platelet adhesion and aggregation. In addition, patagonin did not inhibit contractions of rat aortic smooth muscle induced by high K(+). However, it caused muscular damage to murine gastrocnemius muscle, an action that has not been previously described for any snake venom CRiSPs. Thus, patagonin will be important for studies of the structure-function and evolutionary relationships of this family of proteins that are widely distributed among snake venoms. PMID- 19285569 TI - Practical experience with the maintenance and auditing of a large medical ontology. AB - The Medical Entities Dictionary (MED) has served as a unified terminology at New York Presbyterian Hospital and Columbia University for more than 20 years. It was initially created to allow the clinical data from the disparate information systems (e.g., radiology, pharmacy, and multiple laboratories, etc.) to be uniquely codified for storage in a single data repository, and functions as a real time terminology server for clinical applications and decision support tools. Being conceived as a knowledge base, the MED incorporates relationships among local terms, between local terms and external standards, and additional knowledge about terms in a semantic network structure. Over the past two decades, we have sought to develop methods to maintain, audit and improve the content of the MED, such that it remains true to its original design goals. This has resulted in a complex, multi-faceted process, with both manual and automated components. In this paper, we describe this process, with examples of its effectiveness. We believe that our process provides lessons for others who seek to maintain complex, concept-oriented controlled terminologies. PMID- 19285570 TI - The impact of SNOMED CT revisions on a mapped interface terminology: terminology development and implementation issues. AB - Large-scale mapping efforts have been done in attempts to migrate systems that use proprietary concepts to ones that use terminological standards such as SNOMED CT. As efforts move towards implementation, the target maps should retain a predictable structure including those targets requiring post-coordination of SNOMED CT concepts. In this paper, we compared the editorial guidelines of two versions of SNOMED CT (January 2005 and July 2006, respectively) and noted how the revisions affected a single, comprehensive set of mapped concepts (n=2002) from a legacy system. Changes made to the categories and guidelines for approved attributes were noted and then evaluated against the post-coordinated maps (n=1570) from the original mapping effort. Seventy-one percent (n=1118) of the concepts were affected due to changes made in either SNOMED CT categories or to the revision of approved attributes. While the efforts of each subsequent SNOMED CT version aim for continual improvement, changes made to its core structure and post-coordination guidelines make it more difficult to migrate proprietary data to this reference standard. Attention must be paid to auditing the processes used in terminology development to include the impact that their revisions may have on real-world clinical implementation. PMID- 19285572 TI - Transforming growth factor-beta1 and its receptors in patients with ulcerative colitis. AB - Transforming growth factor-beta1 (TGF-beta1) plays a role in the pathogenesis of ulcerative colitis (UC) by activating its specific receptors (T beta RI-T beta RIII). We investigated the expression of genes encoding for TGF-beta1 and T beta RI-III using RT-QPCR in patients with active and inactive UC and non-IBD controls. The localization and level of TGF-beta1 protein in intestinal tissue was estimated by immunohistochemistry, and serum TGF-beta1 concentrations were determined using ELISA. We found a significant increase in TGF-beta1 gene expression and increase in the expression of genes encoding receptor T beta RI in patients with active UC when compared with controls. The expression of genes encoding T beta RII was found to be higher in patients with both active and inactive UC when compared to controls. Specific staining for TGF-beta1 in fibroblasts was significantly greater in both active and inactive UC as compared to controls. The serum concentration of TGF-beta1 was significantly higher in patients with active UC when compared with controls as well as in UC patients with left side/total colonic extension when compared with those with disease limited to rectum/rectosigmoid area. However, no correlation between TGF-beta1 serum concentrations and UC activity index was found. Increases in TGF-beta1 gene expression and its protein level, associated with altered TGF-beta1 receptor profile indicate a functional role for TGF-beta1 in intestinal inflammatory/repair processes in UC. Increases in TGF-beta1 serum concentrations correlate with extension of disease. PMID- 19285571 TI - A review of auditing methods applied to the content of controlled biomedical terminologies. AB - Although controlled biomedical terminologies have been with us for centuries, it is only in the last couple of decades that close attention has been paid to the quality of these terminologies. The result of this attention has been the development of auditing methods that apply formal methods to assessing whether terminologies are complete and accurate. We have performed an extensive literature review to identify published descriptions of these methods and have created a framework for characterizing them. The framework considers manual, systematic and heuristic methods that use knowledge (within or external to the terminology) to measure quality factors of different aspects of the terminology content (terms, semantic classification, and semantic relationships). The quality factors examined included concept orientation, consistency, non-redundancy, soundness and comprehensive coverage. We reviewed 130 studies that were retrieved based on keyword search on publications in PubMed, and present our assessment of how they fit into our framework. We also identify which terminologies have been audited with the methods and provide examples to illustrate each part of the framework. PMID- 19285573 TI - Complement factors C3a and C5a have distinct hemodynamic effects in the rat. AB - In the rat, C5a infusion mediates well-defined effects including hypotension and neutropenia. Conversely, the comparative effect of C3a in the rat is not yet defined. In the current study, we have investigated C3a receptor (C3aR) activation in the rat, using recombinant human C3a, the C3aR agonist WWGKKYRASKLGLAR, which is a C-terminal analogue of C3a, and a nonpeptide C3aR antagonist SB-290157, as pharmacological tools. In vitro, C3a and WWGKKYRASKLGLAR selectively bound to C3aRs and induced degranulation of C3aR-transfected RBL-2H3 cells. C3a or WWGKKYRASKLGLAR-induced degranulation was dose-dependently antagonized in a surmountable fashion by the nonpeptide C3aR antagonist. Intravenous infusion of C3a and WWGKKYRASKLGLAR to rats induced a rapid, transient and concentration-dependent hypertensive response, which was mediated by C3aR-induced prostanoid release. C3a and WWGKKYRASKLGLAR caused a small drop in circulating neutrophils, but a rise in circulating neutrophils was evident after 90-120 min. In contrast to C3a, C5a infusion resulted in hypotension, and rapid and transient neutropenia. These results demonstrate that C3a and C5a mediate distinct effects on blood pressure and circulating polymorphonuclear leukocytes in the rat. PMID- 19285574 TI - Caffeic acid phenethyl ester inhibits osteoclastogenesis by suppressing NF kappaB and downregulating NFATc1 and c-Fos. AB - Osteoclasts are multinuclear cells of myeloid lineage responsible for bone resorption. The anti-inflammatory property of caffeic acid phenethyl ester (CAPE), an active component of the propolis of honeybee hives, has been revealed. Since the regulatory mechanism of differentiation and activation of osteoclasts shares many well-known signaling pathways with that of inflammation, we investigated whether CAPE has any effect on osteoclastogenesis. CAPE potently suppressed osteoclastogenesis in cultures of bone marrow-derived precursor cells with the osteoclast differentiation factor, receptor activator of nuclear factor kappaB ligand (RANKL). While the RANKL-stimulated activation of the ERK, JNK, and p38 MAPK signaling pathways was not affected, the DNA binding and transcription activity of NF kappaB were reduced by CAPE treatment. In addition, CAPE blocked the induction of NFATc1 and c-Fos following RANKL stimulation. Forced expression of c-Fos could reverse the inhibitory effect of CAPE on osteoclastogenesis. Finally, CAPE significantly inhibited the RANKL-induced osteoclast formation in mouse calvariae in vivo. We propose that CAPE might be useful as a therapeutic agent for treatment of bone destructive diseases. PMID- 19285575 TI - Induction of type 1 cytokines during neem leaf glycoprotein assisted carcinoembryonic antigen vaccination is associated with nitric oxide production. AB - Involvement of the nitric oxide (NO) release in CEAM phi NLGP (carcinoembryonic antigen pulsed macrophages with neem leaf glycoprotein) vaccination and its relationship with vaccine induced type 1 immune response were aimed to study in the present communication. Vaccination with CEAM phi NLGP resulted in macrophage activation as evidenced by its increased number and expression of CD69 marker. Activated macrophages demonstrated upregulation in synthesis of IL-12 and downregulation in IL-10, along with excess IFN gamma production in splenic cells, as evidenced from mRNA analysis. Induction of such type 1 immunity was further confirmed by expression of type 1 specific transcription factor, T-bet and enhancement of intracellular glutathione content. Such vaccination also induced greater nitric oxide (NO) production from macrophages. Dependence of induced type 1 immune response on the NO release and vice versa was studied by in vitro neutralization of IFN gamma/IL-12 and in vivo inhibition of NO production by methylene blue. Obtained results clearly demonstrated the interdependence of two anti-tumor immune functions, namely, NO production and generation of type 1 immune response. Understanding of the mechanism of this NO related immune modulation would have great impact in proposing CEAM phi NLGP vaccine in clinic for the treatment of CEA+ tumors. PMID- 19285576 TI - Novel cytostatic activity of the trypanocidal drug Benznidazole. AB - We have shown that Benznidazole (BZL), a compound with well documented trypanocidal activity, possesses anti-inflammatory properties and inhibits the nuclear factor kappaB (NF-kappaB). Given the relationship between this transcription factor and cell growth, in this study we address the role of NF kappaB blockade by BZL in the proliferation of different cell lines. Our studies demonstrate that this compound significantly reduced proliferation of RAW 264.7 macrophage cell line, as assessed by trypan blue exclusion, MTT reduction and [(3)H]-thymidine incorporation, at a concentration shown to inhibit NF-kappaB. Treatment with BZL also led to growth arrest in CHO, MDCK and HeLa cells. Interestingly, growth inhibition was found to be a reversible process, not accompanied by significant cell death, indicating that the drug behaves mainly as a cytostatic compound. As this effect might be related to NF-kappaB inhibition, we next evaluated whether other NF-kappaB inhibitors could induce growth arrest in RAW 264.7 and HeLa cells. We found that IKK inhibition led to growth arrest in both cell lines, indicating that NF-kappaB inhibition may be the potential mechanism by which BZL inhibits cell proliferation. To the best of our knowledge, this is the first report of an anti-proliferative activity of the trypanocidal drug against different cell lines and provides a mechanistic insight that may help understand some of the adverse effects associated with prolonged treatment. PMID- 19285577 TI - Arrhythmogenic effects of anti-Ro/SSA antibodies on the adult heart: more than expected? AB - The arrhythmogenicity of anti-Ro/SSA antibodies for the foetal heart and their crucial role in the development of congenital heart block is now well established, representing a paradigmatic model of passively acquired autoimmunity. Recently, intriguing data suggest that also the adult heart may represent a possible target of anti-Ro/SSA antibody-mediated autoimmune injury. The prolongation of the QTc interval, possibly resulting from a direct inhibitory interaction between the anti-Ro/SSA antibodies and the potassium current I(Kr) in the heart seems the abnormality more frequently observed in adults with anti Ro/SSA-positive CTD. Although the possibility that anti-Ro/SSA positivity may be considered a risk factor for arrhythmic sudden death in adults has not been demonstrated as yet, preliminary data suggest a relationship among anti-Ro/SSA antibodies, QTc prolongation, and the prevalence of ventricular arrhythmias, also life threatening, in adult patients. PMID- 19285578 TI - Clinical and molecular genetic analysis of a family with macrothrombocytopenia and early onset sensorineural hearing loss. AB - A kindred with inherited macrothrombocytopenia (MTCP) and sensorineural hearing loss (SNHL) from Ghent, Belgium was identified. Currently, joint expression of MTCP and hearing loss are linked to mutations within MYH9 only. Thus, we tested the hypothesis that a mutation within MYH9 is responsible for the autosomal dominant inheritance of MTCP and hearing loss in the Ghent family. A mutation screen of MYH9 coding region including its intron-exon junctions, as well as common hearing loss genes GJB2, GJB3, and GJB6, was performed. However, no pathogenic sequence alteration was identified. Patients' leukocytes were determined to be normal for NMMHC-A distribution via immunofluorescence analysis and free of Dohle body-like inclusions, identified as aggregates of mutant NMHC IIA in MYH9 disorders. Also, western blot analysis with anti-NMHC-IIA antibody identified a single 220 kDa immunoreactive band with normal expression level of NMHC-IIA within the platelets and leukocytes of the affected family members. The immunoblot analysis eliminates the possibility of a large deletion within MYH9 that can escape detection by direct sequencing. Collectively, these results suggest that molecular genetic etiology of the Ghent family disorder may be due to as yet unidentified gene whose mutation(s) yields a phenocopy of the MYH9 related disease. PMID- 19285579 TI - Comparison of risk factors for falls from height between commercial and residential roofers. AB - INTRODUCTION: The purpose of this investigation was to compare commercial roofers and residential roofers in terms of their behaviors, beliefs, working conditions, and attitudes toward the use of fall protection devices, which could lead to fall accidents. METHODS: A cross-sectional sample of 252 roofers participated in the survey in the Midwest (Wisconsin, Illinois, Michigan, Indiana, and Iowa). RESULTS: Residential roofers were more likely to fall (prevalence ratio = 2.28, 95% CI = 1.58, 3.29) [corrected] than commercial roofers. Race/ethnicity, company size, work type, existence [corrected] of fall protection programs, enforcement of fall protection device use, actual use of fall protection devices, years of experience as a roofer, and perceived level of safety at roofing sites were [corrected] significantly associated with fall accidents. IMPACT ON INDUSTRY: This study adds insight into fall accidents from roofs in the construction industry and provides industry-specific cautions against fall accidents that can be reflected in regulatory agency implementation. PMID- 19285580 TI - Dissociation between driving performance and drivers' subjective estimates of performance and workload in dual-task conditions. AB - INTRODUCTION: The current study measured how concurrent driving and in-vehicle activities of different levels of engagement varied in terms of performance and subjective estimates of demand and performance. METHOD: In this test track study, 41 younger and older drivers completed a series of cognitive tasks while driving an instrumented vehicle. One task involved an engaging guessing game where drivers tried to guess the identity of an object. The other task involved a simple mental arithmetic task. RESULTS: We observed some dissociation between drivers' performance and their subjective reports. For instance, drivers tended to estimate their performance as better for the more engaging guessing task than the arithmetic task, though their performance was actually worse. At the same time, subjective estimates of workload across the two tasks did not vary in the dual-task condition even though they did in the single-task baseline conditions, suggesting that drivers failed to account for the added demands in dual-task situations. CONCLUSIONS: We discuss the implications of these findings for driver safety. IMPACT ON INDUSTRY: Crashes due to distraction can carry tremendous costs for employers, in terms of injury, disability, and loss of potentially productive work years, whether these crashes occur on or off the job. PMID- 19285581 TI - Effect of a nurse back injury prevention intervention on the rate of injury compensation claims. AB - PROBLEM: This study evaluated the effect of introducing a No Lifting policy on back injuries to nurses, across an entire health care system. METHODS: Methods included: analysis of the data for all public health agencies in the Australian state of Victoria; compensation data from the Victorian Workcover Authority; data about workforce and program implementation from a retrospective survey of agencies; longitudinal analysis of standardized workers compensation claim rates for back injuries before, during and after the intervention. RESULTS: A statistically significant decline in back injury claim rates during implementation contrasted with no statistically significant trends within the periods before and after the intervention. A statistically significant reduction occurred in mean quarterly standard back injury claim incidence rates per 1,000 equivalent fulltime nursing staff (EFTNS), representing a 24% reduction in standard back injury claims/1000 EFTNS. DISCUSSION: Ergonomics principles encourage changing the work environment to suit the worker. This approach delivered a significant improvement in the immediate term. IMPACT OF INDUSTRY: The substantial decline in back injury rates signifies a major improvement in the safety of a critical aspect of the work environment for nurses. PMID- 19285582 TI - Work injuries and disability. AB - PROBLEM: This study estimated the hazard ratio for disability pension retirement (DPR) for persons who have experienced a work injury causing absence lasting at least one day after the accidental injury occurred and to estimate the fraction of DPR attributable to work injuries. METHODS: A total of 4,217 male and 4,105 female employees from a national survey were followed up for subsequent DPR. RESULTS AND IMPACT ON INDUSTRY AND GOVERNMENT: Having had a work injury was a strong predictor of DPR among men. After control for age, smoking, body mass index, body postures, and physical demands, the hazard ratio (HR) among those employees who had ever experienced a work injury was 1.80 (95% confidence interval (CI): 1.20-2.68). No association was found among women. SUMMARY: Having had a reportable work injury is a strong predictor of subsequent DPR for men. PMID- 19285583 TI - Results from a national survey of Crown prosecutors and defense counsel on impaired driving in Canada: a "System Improvements" perspective. AB - INTRODUCTION: This article summarizes the main findings from a study designed to examine the legal process in Canada as it applies to alcohol-impaired driving from the point of view of Crown prosecutors and defense counsel, and to identify evidentiary or procedural factors that may impact the legal process, the rights of the accused, and interactions of all parts in the legal process. METHOD: The data in this study were collected by means of a survey that was mailed out to the population of Crown prosecutors and defense counsel in Canada. In total, 765 prosecutors and 270 defense lawyers or an estimated 33% of all Canadian prosecutors and 15% of defense lawyers completed and returned the questionnaire. The "systems improvement" paradigm was used to interpret the findings and draw conclusions. Such an approach acknowledges the importance of the context in which countermeasures are implemented and delivered and the structures or entities used to deliver countermeasures to a designated target group. RESULTS: Results on type of charges and breath alcohol concentration, caseload, case outcomes, case preparation time, conviction rate at trial and overall conviction rate, reasons for acquittals and time to resolve cases are described. DISCUSSION: The findings from this national survey suggest that there are important challenges within the criminal justice system that impede the effective and efficient processing of impaired driving cases. Some of these challenges occur as a function of practices and policies, while others occur as a function of legislation. IMPACT ON INDUSTRY: This study illustrates that a "system improvements" approach that acknowledges the importance of all elements of the criminal justice system and the interaction between those elements, can be beneficial in overcoming the alcohol-impaired driving problem. PMID- 19285584 TI - Social norms of accompanied young children and observed crossing behaviors. AB - INTRODUCTION: Social norms for accompanied young children and crossing behaviors were examined in two studies conducted in an Ultra-Orthodox Jewish community in Israel. METHOD: In Study 1, road behaviors of young children crossing with and without accompaniment and older children were observed, and the actual social norm for accompanied school children younger than 9-years-old was examined. In Study 2, the perceived norm of accompaniment was tested by questionnaires. RESULTS: Young children who crossed without accompaniment exhibited poorer crossing skills compared to older children and to young children crossing with accompaniment. In the four locations observed, the actual accompaniment rate ranged between 15%-60%. The perceived social norm for child accompaniment was lower than the actual norm. CONCLUSIONS: The discussion refers to both theoretical issues and their practical implications. PMID- 19285585 TI - An economic evaluation of a participatory ergonomics process in an auto parts manufacturer. AB - PROBLEM: We assess the costs and consequences of a participatory ergonomics process at a Canadian car parts manufacturer from the perspective of the firm. METHOD: Regression modeling was used with interrupted time series data to assess the impact of the process on several health measures. Consequences were kept in natural units for cost-effectiveness analysis, and translated into monetary units for cost-benefit analysis. RESULTS: The duration of disability insurance claims and the number of denied workers' compensation claims was significantly reduced. The cost-effectiveness ratio is $12.06 per disability day averted. The net present value is $244,416 for a 23-month period with a benefit-to-cost ratio of 10.6, suggesting that the process was worth undertaking (monetary units in 2001 Canadian dollars). DISCUSSION: Our findings emphasize the importance of considering a range of outcomes when evaluating an occupational health and safety intervention. IMPACT ON INDUSTRY: Participatory ergonomics process can be cost effective for a firm. PMID- 19285586 TI - Attitudes toward a primary seat belt law among adults in a rural state with a secondary seat belt law. AB - PROBLEM: Enforced primary seatbelt laws can reduce morbidity and mortality associated with motor-vehicle crashes. Constituent support is an important factor associated with legislator voting behavior toward injury prevention laws. Little is known about attitudes toward a primary seat belt law among adults in rural states without a primary seat belt law. METHODS: Data from the Behavioral Risk Factor Surveillance System (BRFSS) survey, a telephone survey of a representative sample of adults in Montana, were used to assess attitudes toward a primary seat belt law. RESULTS: Sixty-one percent of respondents supported a primary seat belt law. Using multiple logistic regression analyses, women (AOR 1.87; 95% CI 1.49 2.36), persons aged 65 years and older (1.45; 1.06-1.96), American Indians (2.71; 1.55-4.75), those with health insurance (1.51; 1.07-2.14), and those who reported always wearing their seat belt (4.05; 3.14-5.21) were more likely to support a primary seat belt law than respondents without these characteristics. CONCLUSIONS: The majority of adults in a rural state support a primary seat belt law. PMID- 19285587 TI - "Safety is everyone's job:" the key to safety on a large university construction site. AB - PROBLEM: Construction risk management is challenging. METHOD: We combined data on injuries, costs, and hours worked, obtained through a Rolling Owner-Controlled Insurance Program (ROCIP), with data from focus groups, interviews, and field observations, to prospectively study injuries and hazard control on a large university construction project. RESULTS: Lost-time injury rates (1.0/200,000 hours worked) were considerably lower than reported for the industry, and there were no serious falls from height. Safety was considered in the awarding of contracts and project timeline development; hazard management was iterative. A top-down management commitment to safety was clearly communicated to, and embraced by, workers throughout the site. DISCUSSION AND IMPACT: A better understanding of how contracting relationships, workers' compensation, and liability insurance arrangements influence safety could shift risk management efforts from worker behaviors to a broader focus on how these programs and relationships affect incentives and disincentives for workplace safety and health. PMID- 19285588 TI - An epidemiologic comparison of injuries presenting to a pediatric emergency department and local urgent care facilities. AB - PROBLEM: The objective of this study was to compare the epidemiology of injuries presenting to emergency department (ED) and urgent care (UC) facilities of a single, NEISS-affiliated hospital. METHOD: Patient medical records (n=36,811) were used to compare injury incidence, injury characteristics, and demographic characteristics between the ED, on-site UC, and off-site UC during 2006. RESULTS: ED presentations were more likely to be open wounds and motor vehicle-related compared to on-site UC presentations. ED presentations were more likely to be system wide/late effects, be made by an African American, or be paid through Medicaid compared to off-site UC presentations. On-site UC presentations were more likely to be made by an African American or be paid through Medicaid compared to off-site UC presentations. DISCUSSION: ED and UC injury characteristics and patient demographics differ. With no nationally representative UC injury surveillance, current research likely underestimates injury incidence and presents skewed profiles. IMPACT ON INDUSTRY: This article adds insight into the generalizability of ED-based injury surveillance to UC injuries. PMID- 19285589 TI - Unintentional childhood injuries in the United States: key findings from the CDC childhood injury report. PMID- 19285590 TI - Early Miocene catarrhine dietary behaviour: the influence of the Red Queen Effect on incisor shape and curvature. AB - The early Miocene catarrhine fossil record of East Africa represents a diverse and extensive adaptive radiation. It is well accepted that these taxa encompass a dietary range similar to extant hominoids, in addition to some potentially novel dietary behaviour. There have been numerous attempts to infer diet for these taxa from patterns of dental allometry and incisor and molar microwear, however, morphometric analyses until now have been restricted to the post-canine dentition. It has already been demonstrated that given the key functional role of the incisors in pre-processing food items prior to mastication, there is a positive correlation between diet and incisal curvature (Deane, A.S., Kremer, E.P., Begun, D.R., 2005. A new approach to quantifying anatomical curvatures using High Resolution Polynomial Curve Fitting (HR-PCF). Am. J. Phys. Anthropol. 128(3), 630-638.; Deane, A.S., 2007. Inferring dietary behaviour for Miocene hominoids: A high-resolution morphometric approach to incisal crown curvature. Ph.D. Dissertation. The University of Toronto.). This study seeks to re-examine existing dietary hypotheses for large-bodied early Miocene fossil catarrhines by contrasting the incisal curvature for these taxa with comparative models derived from prior studies of the correlation between extant hominoid incisor curvature and feeding behaviour. Incisor curvature was quantified for 78 fossil incisors representing seven genera, and the results confirm that early Miocene fossil catarrhines represent a dietary continuum ranging from more folivorous (i.e., Rangwapithecus) to more frugivorous (i.e., Proconsul) diets, as well as novel dietary behaviours that are potentially similar to extant ceboids (i.e., Afropithecus). Additionally, early Miocene fossil catarrhine incisors are less curved than extant hominoid incisors, indicating a general pattern of increasing mesio-distal and labial curvature through time. This pattern of morphological shifting is consistent with the Red Queen Effect (Van Valen, L., 1973. A new evolutionary law. Evol. Theory 1, 1-30), which predicts that taxa that are removed from one another by geological time, although potentially having similar diets, may exhibit differing degrees of a similar dietary adaptation (i.e., differing degrees of incisal curvature). PMID- 19285591 TI - Shells and ochre in Middle Paleolithic Qafzeh Cave, Israel: indications for modern behavior. AB - Qafzeh Cave, the burial grounds of several anatomically modern humans, producers of Mousterian industry, yielded archaeological evidence reflecting their modern behavior. Dated to 92ka BP, the lower layers at the site contained a series of hearths, several human graves, flint artifacts, animal bones, a collection of sea shells, lumps of red ochre, and an incised cortical flake. The marine shells were recovered from layers earlier than most of the graves except for one burial. The shells were collected and brought from the Mediterranean Sea shore some 35km away, and are complete Glycymeris bivalves, naturally perforated. Several valves bear traces of having been strung, and a few had ochre stains on them. PMID- 19285592 TI - In the book of life, the answers aren't in the back--Charlie Brown, fictional character of the Peanuts comic strip created by Charles Schulz. PMID- 19285593 TI - ICRP Publication 107. Nuclear decay data for dosimetric calculations. AB - In this report, the Commission provides an electronic database of the physical data needed in calculations of radionuclide-specific protection and operational quantities. This database supersedes the data of Publication 38 (ICRP, 1983), and will be used in future ICRP publications of dose coefficients for the intake of or exposure to radionuclides in the workplace and the environment.The database contains information on the half-lives, decay chains, and yields and energies of radiations emitted in nuclear transformations of 1252 radionuclides of 97 elements. The CD accompanying the publication provides electronic access to complete tables of the emitted radiations, as well as the beta and neutron spectra. The database has been constructed such that user-developed software can extract the data needed for further calculations of a radionuclide of interest. A Windows-based application is provided to display summary information on a user specified radionuclide, as well as the general characterisation of the nuclides contained in the database. In addition, the application provides a means by which the user can export the emissions of a specified radionuclide for use in subsequent calculations. PMID- 19285596 TI - Indonesian women of childbearing age are at greater risk of clinical vitamin A deficiency in families that spend more on rice and less on fruits/vegetables and animal-based foods. AB - Clinical vitamin A deficiency is characterized by night blindness and greater morbidity and mortality. The aim of this study was to examine the relationships between household food expenditures and night blindness among nonpregnant women of childbearing age among families in the slums of Jakarta, Indonesia. In a cross sectional study of 42 974 households in the Indonesian Nutrition Surveillance System, 1998 to 2003, night blindness was assessed in nonpregnant women. Food expenditures were divided into 5 major categories as follows: plant-based foods (fruits and vegetables), animal-based foods, eggs, other nongrain foods, and grain foods (primarily rice), calculated as percentage of total weekly per capita food expenditure, and expressed in quintiles. The proportion of households with night blindness in nonpregnant women was 0.72%. Plant-based food, animal-based food, and eggs were associated with reduced odds of night blindness (odds ratio [OR], 0.47; 95% confidence interval [CI], 0.33-0.67; P < .0001, and OR, 0.47; 95% CI, 0.29-0.76; P = .002; OR, 0.62; 95% CI, 0.44-0.85; P = .004), respectively, among families in the highest compared with the lowest quintile, adjusting for potential confounders. Grain food expenditures were associated with increased odds of night blindness among nonpregnant women (OR, 2.80; 95% CI, 1.86-4.22; P < .0001) among families in the highest compared with the lowest quintile, adjusting for potential confounders. This study suggests that nonpregnant women are at greater risk of clinical vitamin A deficiency where families spend more on rice and less on animal and plant-based foods, a situation that is more typical when food prices are high. PMID- 19285597 TI - Zinc levels in seminal plasma are associated with sperm quality in fertile and infertile men. AB - Zinc has antioxidative properties and plays an important role in scavenging reactive oxygen species. We hypothesized that in the absence of Zn, the possibility of increased oxidative damage exists that would contribute to poor sperm quality. Therefore, measurement of seminal Zn in the seminal plasma of males with a history of subfertility or idiopathic infertility is necessary and can be helpful in fertility assessment. The primary objective of the present study was to assess the relationship between Zn levels in seminal plasma with sperm quality in fertile and infertile men. Semen samples were provided by fertile (smoker [n = 17], nonsmoker [n = 19]) and infertile men (smoker [n = 15], nonsmoker [n = 21]). After semen analysis, concentrations of Zn, Mg, Ca, Na, and K in the seminal plasma of all groups were determined by atomic absorption spectroscopy. Element concentrations in seminal plasma of all groups were in the order Na > K > Ca > Zn > Mg. Fertile subjects, smoker or not, demonstrated significantly higher seminal Zn levels than any infertile group (P < .001). A trend was observed for a lower Zn levels in seminal plasma of smokers compared with nonsmokers. Seminal Zn in fertile and infertile (smokers or nonsmokers) males correlated significantly with sperm count (P < .01) and normal morphology of sperm (P < .001). There was a significantly positive correlation between seminal Zn with Ca (P < .01) and K (P < .01) levels in all specimens. In conclusion, poor Zn nutrition may be an important risk factor for low quality of sperm and idiopathic male infertility. PMID- 19285598 TI - A snack enriched with oral branched-chain amino acids prevents a fall in albumin in patients with liver cirrhosis undergoing chemoembolization for hepatocellular carcinoma. AB - Nutritional support may play an important role in management of liver cirrhosis (LC) associated with unresectable hepatocellular carcinoma (HCC). Total protein and albumin deteriorate in patients with LC undergoing trans-arterial chemoembolization (TACE). Therefore, in this study, we examined the hypothesis that short-term administration of branched-chain amino acids (BCAA) will prevent a fall in total protein and albumin in the perioperative period. The subjects were 56 patients who underwent TACE for HCC between 2004 and 2005 at Nagasaki University Hospital. The patients were randomly placed in the BCAA group (n = 28) or a control group (n = 28). The patients in the BCAA group consumed a snack containing 50 g of BCAA once a day at 10:00 pm starting 1 day before TACE and continuing until 2 weeks after TACE. A comparison of baseline and end point data showed greater decreases in the concentrations of total protein, albumin, cholinesterase, and total cholesterol and in the red blood cell count in the control group compared to the BCAA group. Ammonia levels decreased in the BCAA group and increased in the control group. Our findings indicate that a BCAA supplement taken orally as a late evening snack prevents suppression of liver function by TACE in patients with LC complicated with HCC during the 2-week period after TACE. PMID- 19285599 TI - Depressed serum selenoprotein P: possible new predicator of increased risk for cerebrovascular events. AB - Selenium protection against cellular damage by oxygen radicals is accomplished through selenoproteins. Thus, selenium protection during the development of stroke, an oxidative stress-related disease, may not be appropriately reflected in the total serum selenium concentration. Therefore, we hypothesized that serum selenoproteins should also be measured to understand the relationship between selenium status and oxidative stress. To establish whether stroke is associated with changes in serum selenoprotein levels, a population-based, nested case control study was performed. The subjects were recruited from 1632 residents older than 40 years who had completed health examinations in 1992. Blood samples collected from 30 controls and 30 initial stroke victims between 1992 and 1994 were analyzed for total serum selenium and selenium-containing protein distribution. Selenium-containing proteins were separated using 2 high performance liquid chromatography columns in tandem and detected by inductively coupled plasma-mass spectrometry. The mean serum selenium concentration was lower in the patients who had a stroke than in the controls (105.2 vs 116.5 microg/L). Selenium contents in glutathione peroxidase and albumin did not show any significant difference; however, selenoprotein P was significantly lower in the stroke cases than in the controls (54.5 vs 63.0 microg/L, P = .006). Results from multivariate logistic regression analysis showed that reduced serum level of selenoprotein P was associated with a higher risk of stroke (odds ratio = 0.28; 95% confidence interval, 0.10-0.85). PMID- 19285600 TI - Greater satiety response with resistant starch and corn bran in human subjects. AB - Some studies suggest high-fiber foods are more satiating than foods with little or no fiber. However, we hypothesized that certain types of dietary fiber may enhance satiety more than others. Healthy men and women (N = 20) participated in this acute, randomized double-blind, crossover study comparing the effects of 4 fibers and a low-fiber (LF) treatment on satiety. On 5 separate visits, fasting subjects consumed either a LF muffin (1.6 g fiber) or 1 of 4 high-fiber muffins (8.0-9.6 g fiber) for breakfast. The subjects used 4 questions on 100 mm visual analogue scales to rate satiety at baseline and at regular intervals for 180 minutes after muffin consumption. Responses were analyzed as area under the curve and significant differences from baseline. Satiety differed among treatments. Resistant starch and corn bran had the most impact on satiety, whereas polydextrose had little effect and behaved like the LF treatment. Results from this study indicate that not all fibers influence satiety equally. PMID- 19285601 TI - G allele at RAGE SNP82 is associated with proinflammatory markers in obese subjects. AB - Obesity is closely associated with low-grade inflammation. The Gly82Ser (G82S) polymorphism in the receptor for the advanced glycation end products (RAGE) gene related to RAGE expression is also involved in inflammatory response. We examined the association between RAGEG82S and obesity on soluble RAGE (sRAGE) and inflammatory markers in Korean men. The following were measured: anthropometric and biochemical parameters, RAGEG82S polymorphism, sRAGE, advanced glycation end products (AGEs), and inflammatory markers in men (n = 1252; range, 30-70 years; body mass index [BMI], > or =18.5 kg/m(2)). Allele frequencies satisfied Hardy Weinberg Equilibrium (G/G: 72.2%, G/S: 25.5%, S/S: 2.3%). RAGEG82S (beta coefficient = -0.384, P < .001) and BMI (beta-coefficient = -0.168, P = .001) were major factors affecting sRAGE concentrations. In all subjects, those with 'S/S' homozygotes showed the lowest levels of sRAGE (G/G: 1036.3 +/- 40.3, G/S: 807.0 +/- 49.6, S/S: 443.0 +/- 47.8 pg/mL) before (P < .001) and after adjusted for age, BMI, cigarette smoking, and alcohol drinking (P < .001). When subdivided according to BMI of 25 kg/m(2) (Asian Pacific guideline), obese subjects (BMI > or =25 kg/m(2)) had significantly lower levels of sRAGE (831.7 +/- 36.7 vs 1022.7 +/- 47.8 pg/mL, P = .009) and higher levels of high sensitivity C-reactive protein (hs-CRP) (1.10 +/- 0.07 vs 0.72 +/- 0.05 mg/dL, P < .001) compared with nonobese subjects (BMI <25 kg/m(2)). Particularly in obese subjects, S/S carriers showed significantly higher concentrations of AGEs (P = .012) and hs-CRP (P = .006) than G allele carriers, whereas nonobese people had no significant RAGEG82S related differences in AGEs (P = .743) and hs-CRP (P = .436). In conclusion, G allele at RAGEG82S may be more associated with inflammatory markers under obese status than nonobese conditions. In this case, it may help to suggest proper dietary modification for controlling obesity to people with genetic variants. PMID- 19285602 TI - Vitamin C attenuates hypochlorite-mediated loss of paraoxonase-1 activity from human plasma. AB - Paraoxonase 1 (PON1) is a cardioprotective enzyme associated with high-density lipoprotein (HDL). We tested the hypothesis that vitamin C protects HDL and PON1 from deleterious effects of hypochlorous acid, a proinflammatory oxidant. In our experiments, HDL (from human plasma) or diluted human plasma was incubated with hypochlorite in either the absence (control) or presence of vitamin C before measuring chemical modification and PON1 activities. Vitamin C minimized chemical modification of HDL, as assessed by lysine modification and accumulation of chloramines. In the absence of vitamin C, chloramines accumulated to 114 +/- 4 micromol/L in HDL incubated with a 200-fold molar excess of hypochlorite; but addition of vitamin C (200 micromol/L) limited formation to 36 +/- 6 micromol/L (P < .001). In plasma exposed to hypochlorite, IC(50) values of 1.2 +/- 0.1, 9.5 +/- 1.0, and 5.0 +/- 0.6 mmol/L were determined for PON1's phosphotriesterase, arylesterase, and (physiologic) lactonase activities, respectively. Vitamin C lessened this inhibitory effect of hypochlorite on PON1 activities. In plasma supplemented with vitamin C (400 micromol/L), PON1 phosphotriesterase activity was 72% +/- 17% of normal after incubation with hypochlorite (2 mmol/L), compared with 42% +/- 6% for unsupplemented plasma (P < .05). Similar effects were seen for other PON1 activities. In some experiments, vitamin C also appeared to reverse hypochlorite-mediated loss of PON1 phosphotriesterase activity; but this effect was not observed for the other PON1 activities. In conclusion, vitamin C attenuated hypochlorite-mediated loss of PON1 activity in vitro and may, therefore, preserve cardioprotective properties of HDL during inflammation. PMID- 19285603 TI - Masou salmon (Oncorhynchus masou) ethanol extract decreases 3-hydroxy-3 methylglutaryl coenzyme A reductase expression in diet-induced obese mice. AB - This study was designed to evaluate the hypocholesterolemic effects of masou salmon 70% ethanol extract (MSE) and to determine the molecular mechanism by which MSE exerts its effects in high-fat (HF) diet-induced obese mice. We hypothesize that the MSE may contain abundant n-3 fatty acids, so a diet containing MSE may also have hypolipidemic effects by assessing several key gene expressions in cholesterol metabolism such as the low-density lipoprotein (LDL) receptor, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, and cholesterol 7alpha-hydroxylase (CYP7A1). To test this hypothesis, C57BL/6J mice were fed a 40% HF diet for 5 weeks, after which time the animals were fed an HF diet containing 0 mg/kg, 75 mg/kg, or 150 mg/kg MSE (HF, HF + MSE 1, and HF + MSE 2 groups, respectively) for an additional 4 weeks (n = 8 in each group, for a total of 24 mice). We found that feeding MSE with an HF diet prevented hypercholesterolemia in diet-induced obese mice; daily MSE feeding reduced total cholesterol levels in plasma and liver by 12.3% and 16.2%, respectively. Furthermore, we examined the expression of key cholesterol metabolism genes by reverse transcription-polymerase chain reaction and found that messenger RNA levels of HMG-CoA reductase were decreased by up to 5-fold, but the expression of both LDL receptor and CYP7A1 did not change. Thus, MSE may exert its hypocholesterolemic effect by altering the expression of HMG-CoA reductase. PMID- 19285604 TI - Diets containing blueberry extract lower blood pressure in spontaneously hypertensive stroke-prone rats. AB - Oxidative stress in the vasculature and kidneys contributes to hypertension, a major risk factor for cardiovascular disease. Blueberries (BB) are rich in antioxidants, and so we hypothesized that feeding diets enriched with BB would slow the development of hypertension in spontaneously hypertensive stroke-prone rats (SHRSP). Eight-week-old normotensive rats and SHRSP were fed either a control diet (Con) or a diet enriched with 3% freeze-dried BB for 8 weeks. Systolic blood pressure (SBP) was measured at weeks 2, 4, 6, 7, and 8 by the tail cuff method, and urine was collected at weeks 4 and 8. The SBP was elevated in SHRSP relative to normotensive rats over the entire 8-week feeding period. In SHRSP consuming BB, SBP was 19% lower at week 4 and 30% lower at week 6, relative to SHRSP on Con. Maximum SBP was 216 +/- 11 mm Hg in SHRSP consuming Con vs 178 +/- 15 mm Hg in the BB-fed group (P = .036). Spontaneously hypertensive stroke prone rats had elevated levels of urine F2-isoprostanes/creatinine relative to normotensive rats, indicating systemic oxidative stress in this strain. Blueberry feeding had no effect on urinary excretion of F2-isoprostanes; therefore, it is unlikely that a systemic antioxidant effect of BB is responsible for the antihypertensive effects at weeks 4 and 6. Blueberry-fed rats had reduced markers of renal oxidative stress, such as proteinuria and kidney nitrites. Thus, a 3% BB diet may be capable of protecting the kidneys from oxidative damage in SHRSP, thereby reducing the magnitude of hypertension. PMID- 19285605 TI - Low-dose supplementation with active hexose correlated compound improves the immune response to acute influenza infection in C57BL/6 mice. AB - Supplementation with mushroom-derived active hexose correlated compound (AHCC) modulates immunity and increases survival in response to a broad spectrum of acute infections, including influenza virus infection. However, dose-response data are nonexistent. Therefore, the aims of this study were to evaluate AHCC supplementation at various doses and determine the effects of low-dose supplementation on the immune response in a mouse model of influenza virus infection. We hypothesized that AHCC supplementation would influence the immune response to influenza infection in a dose-dependent manner. Male C57BL/6 mice were supplemented with AHCC at daily doses of 0.05, 0.1, 0.5, and 1 g/kg and infected intranasally with influenza A virus (H1N1, PR8). Supplemented mice demonstrated a dose-dependent increase in survival and reduction in the loss of body weight. To further evaluate the effects of low-dose AHCC supplementation on the immune response to influenza infection, mice were supplemented with 0.1 g/kg per day and infected with a sublethal dose of influenza virus. Supplemented mice exhibited enhanced virus clearance and decreased weight loss compared to controls. Low-dose supplementation did not influence total natural killer (NK) cell cytotoxicity, although lytic efficiency was increased in the spleens of AHCC supplemented mice, indicating enhanced NK cell function per cell. In conclusion, these data suggest that the effects of AHCC on the immune response to influenza infection are dose dependent and that low-dose AHCC supplementation improves the response to influenza infection despite no effect on total NK cell cytotoxicity. PMID- 19285606 TI - Executive summary: Kidney Early Evaluation Program (KEEP) 2008 annual data report. PMID- 19285607 TI - Diabetes mellitus and CKD awareness: the Kidney Early Evaluation Program (KEEP) and National Health and Nutrition Examination Survey (NHANES). AB - BACKGROUND: Diabetes contributes to increased morbidity and mortality in patients with chronic kidney disease (CKD). We sought to describe CKD awareness and identify factors associated with optimal glycemic control in diabetic and nondiabetic individuals both aware and unaware of CKD. METHODS: This cross sectional analysis compared Kidney Early Evaluation Program (KEEP) and National Health and Nutrition and Examination Survey (NHANES) 1999 to 2006 participants with diabetes and CKD. CKD was defined and staged using glomerular filtration rate (estimated by using the 4-variable Modification of Diet in Renal Disease Study equation) and urine albumin-creatinine ratio. NHANES defined diabetes as self-reported diabetes or fasting plasma blood glucose level of 126 mg/dL or greater, and KEEP as self-reported diabetes or diabetic retinopathy, use of diabetes medications, fasting blood glucose level of 126 mg/dL or greater, or nonfasting glucose level of 200 mg/dL or greater. RESULTS: Of 77,077 KEEP participants, 20,200 (26.2%) were identified with CKD and 23,082 (29.9%) were identified with diabetes. Of 9,536 NHANES participants, 1,743 (18.3%) were identified with CKD and 1,127 (11.8%) were identified with diabetes. Of KEEP participants with diabetes and CKD (n = 7,853), 736 (9.4%) were aware of CKD. Trends in lack of CKD awareness were similar for KEEP participants with and without diabetes. Unaware participants with and without diabetes identified with stages 1 and 2 CKD were less likely to reach target glucose levels, defined as fasting glucose level less than 126 mg/dL or nonfasting glucose level less than 140 mg/dL, than those with stages 3 to 5 (odds ratio, 0.69; 95% confidence interval, 0.62 to 0.78; odds ratio, 0.69; 95% confidence interval, 0.58 to 0.81; P < 0.001, respectively). CONCLUSION: Our data support that KEEP, as a targeted screening program, is a more enriched population with CKD and comorbid diabetes than NHANES. In addition, our findings highlight the relationship between dysglycemia and early stages of unidentified CKD. PMID- 19285608 TI - Hypertension in early-stage kidney disease: an update from the Kidney Early Evaluation Program (KEEP). AB - BACKGROUND: Chronic kidney disease (CKD) is a worldwide public health problem. Systolic blood pressure as an associated feature of CKD has not been fully explored in community volunteer and nationally representative samples of the US population. METHODS: This cross-sectional analysis evaluated hypertension and early-stage CKD in participants in the Kidney Early Evaluation Program (KEEP), a voluntary community-based health screening program administered by the National Kidney Foundation, and the National Health and Nutrition Examination Survey (NHANES) data to assess similarities and differences between these populations. Participants in both databases were 18 years or older. RESULTS: The KEEP database included 88,559 participants and the NHANES included 20,095. Hypertension prevalence was greater in KEEP (69.6%) than NHANES (38.1%; P < 0.001). Compared with NHANES participants, KEEP participants had greater rates of obesity (79.5% versus 51.5%; P < 0.001) and diabetes (28.0% versus 8.9%; P < 0.001). In participants with diabetes, KEEP had slightly greater rates of prevalent hypertension (88.5% versus 85.7%; P = 0.03). In participants with hypertension, CKD stages 3 and 4 were more prevalent in KEEP than NHANES (79.1% versus 69.3%; P < 0.001). Rates of CKD stages 3 and 4 were greater in KEEP than NHANES for the following subgroups: African Americans (72.4% versus 57.4%; P < 0.001), smokers (69.1% versus 55.6%; P = 0.002), and participants with hypercholesterolemia (80.2% versus 71.9%; P < 0.001). CONCLUSIONS: In the volunteer KEEP population, rates of hypertension and CKD were greater than in NHANES, most prominently in African Americans and participants with increased cardiovascular risk. PMID- 19285609 TI - Plasma parathyroid hormone level and prevalent cardiovascular disease in CKD stages 3 and 4: an analysis from the Kidney Early Evaluation Program (KEEP). AB - BACKGROUND: Cardiovascular disease (CVD) is the most common cause of death in patients with chronic kidney disease (CKD). Secondary hyperparathyroidism is common in patients with CKD, and its relationship to CVD is not well defined. This analysis aims to assess whether serum intact parathyroid hormone (PTH) level is an independent risk factor for CVD in patients with CKD stages 3 and 4. METHODS: In this cross-sectional study, medical history surveys, including CVD events, were collected from 4,472 patients with stages 3 and 4 CKD identified by the National Kidney Foundation Kidney Early Evaluation Program (KEEP), which included blood pressure measurement and laboratory testing. Age, hemoglobin level, estimated glomerular filtration rate, serum phosphorus level, and serum calcium level were evaluated as continuous variables, and plasma PTH levels, by tertile: less than 35, 35 to 70, and greater than 70 pg/mL. Multivariate logistic regression was used to estimate odds ratios (ORs) of CVD predictor variables. RESULTS: Mean age was 68.3 +/- 11.8 years. Of the study population, 68% were women, 69% were white, 6% were current smokers, 45% were obese, 46% had diabetes, and 83% had hypertension. A history of CVD was present for 1,972 (44.1%), and plasma PTH level greater than 70 pg/mL, for 2,239 (50.1%). Multivariate logistic regression showed ORs for CVD events increasing with age (OR, 1.03; P < 0.001), male sex (OR, 1.51; P < 0.001), diabetes (OR, 1.73; P < 0.001), hypertension (OR, 1.43; P < 0.001), and intact PTH level greater than 70 pg/mL (OR, 1.51; P < 0.001; reference, <35 pg/mL). CONCLUSIONS: PTH level greater than 70 pg/mL is independently associated with CVD events in patients with CKD stages 3 and 4. No association was observed between serum phosphorus or calcium level and CVD events. These findings provide support for intact PTH testing, along with testing for other indicators of CKD mineral and bone disorders, at earlier CKD stages. PMID- 19285610 TI - Changing leadership for the journal of heart and lung transplantation. PMID- 19285612 TI - Neuromuscular diseases after cardiac transplantation. AB - BACKGROUND: Cardiac transplantation is a therapeutic option in end-stage heart failure. Peripheral nervous system (PNS) disease is known to occur in cardiac transplant recipients but has not been fully characterized. METHODS: This retrospective cohort review reports the PNS-related concerns of 313 cardiac transplant recipients (28% women, 8% children) over an 18-year period at a single institution. RESULTS: Thirty percent of patients (95 of 313) reported a PNS related concern in the post-transplant period, but only 5% had a concern in the first 6 weeks after transplant. The relative frequency of PNS-related complaints was as follows: polyneuropathy, 33%; muscle disease, 26%; mononeuropathy, 17%; radiculopathy, 13%; small-fiber polyneuropathy, 4%; plexopathy, 3%; and other (e.g., herpes zoster), 4%. CONCLUSIONS: Etiology of these concerns can be divided into four broad categories: (1) immediate post-operative complications (e.g., brachial plexus stretch injury); (2) concerns related to the underlying disease prompting transplantation (e.g., polyneuropathy secondary to amyloidosis); (3) concerns related to necessary medications (e.g., steroid-associated myopathy); and (4) concerns reflective of aging in a post-transplant population with enhanced survival (e.g., degenerative joint disease-related radiculopathy). PMID- 19285613 TI - Can the Seattle heart failure model be used to risk-stratify heart failure patients for potential left ventricular assist device therapy? AB - BACKGROUND: According to results of the REMATCH trial, left ventricular assist device therapy in patients with severe heart failure has resulted in a 48% reduction in mortality. A decision tool will be necessary to aid in the selection of patients for destination left ventricular assist devices (LVADs) as the technology progresses for implantation in ambulatory Stage D heart failure patients. The purpose of this analysis was to determine whether the Seattle Heart Failure Model (SHFM) can be used to risk-stratify heart failure patients for potential LVAD therapy. METHODS: The SHFM was applied to REMATCH patients with the prospective addition of inotropic agents and intra-aortic balloon pump (IABP) +/- ventilator. RESULTS: The SHFM was highly predictive of survival (p = 0.0004). One-year SHFM-predicted survival was similar to actual survival for both the REMATCH medical (30% vs 28%) and LVAD (49% vs 52%) groups. The estimated 1-year survival with medical therapy for patients in REMATCH was 30 +/- 21%, but with a range of 0% to 74%. The 1- and 2-year estimated survival was 1 year post-OHT in 13% of patients. The average EMB score was significantly higher in QE(+) biopsies. A comparison of the two groups revealed a significantly higher number of AR episodes and number of patients with at least one episode of AR in QE(+) patients. There was no significant difference in number of CAV occurrences between groups. CONCLUSIONS: The QE seems to be a marker of the same increased immune system activity that can lead to AR. A relationship between QE and CAV was not supported by the present results. PMID- 19285618 TI - Use of hearts transplanted from donors with severe sepsis and infectious deaths. AB - BACKGROUND: The reluctance to use organs from donors who have died from severe infections is based on the potential transmission of an infectious agent to the recipient and on the uncertainty about allograft function in the setting of severe donor sepsis. METHODS: From 1999 to 2007, donor hospital records were reviewed which focused on microbiology cultures and sensitivity results; type and duration of antimicrobial therapy; hemodynamic data, results of echocardiogram, and imaging studies. Preliminary positive and negative results from pre-harvest blood, respiratory, urine, and cerebrospinal fluid cultures were verified with the procurement agency. The harvesting surgeon performed gross inspection of donor valvular structures. RESULTS: Nine donor hearts were transplanted from patients who expired from community onset infections with severe septic shock, meningitis, and/or pneumonia caused by Streptococcus pneumoniae (n = 4), Streptococcus milleri (n = 2), Neisseria meningitidis (n = 2), and unidentified gram- positive cocci (n = 1). Four donors had probable infection-induced intracranial hemorrhage, and all donors were vasopressor-dependent before organ procurement. No evidence of donor-transmitted infection, sepsis, or rejection was observed, and long-term function remained excellent; allograft dysfunction in three patients resolved after transplant. Our series of nine donors represents approximately 1.3% of successfully transplanted cardiac allografts during the respective period of review. CONCLUSIONS: Patients succumbing to severe infections (meningitis, pneumonia, and septic shock) should not be arbitrarily excluded for possible heart donation. Assessing the suitability of donors with severe infections requires flawless communication between the donor and transplant facility, including a comprehensive evaluation of donor infection and pathogen(s), severity of sepsis, adequacy of antimicrobial treatment, and the degree of sepsis-induced myocardial dysfunction. PMID- 19285619 TI - Circulatory load during hypoxia impairs post-transplant myocardial functional recovery in donation after cardiac death. AB - BACKGROUND: Circulatory load during hypoxia is unavoidable in donation after cardiac death (DCD) hearts, but it causes severe myocardial damage. The impact of circulatory load on donor heart function has not been investigated. The purpose of this study was to evaluate its effect on post-transplant functional recovery of DCD hearts. METHODS: Twelve donor pigs (20 kg) were used. Cardiac arrest was induced by asphyxiation (turning off the ventilator) in the load group (n = 6) and by exsanguination (dividing the vena cava) in the unload group (n = 6). Left ventricle end-diastolic volume (LDEDV) and end-systolic pressure (LVESP) were monitored until cardiac arrest. Orthotopic transplantation was performed after 30 minute warm ischemia following cardiac arrest. After weaning from cardiopulmonary bypass, left ventricular end-diastolic pressure-volume ratio (LV Emax) and creatine kinase (CK-MB) were measured while on 0.1 microg/kg/min epinephrine. RESULTS: During the agonal period, the maximum LVEDV and LVESP in the load group were 132 +/- 1% of baseline at 10 minutes and 148 +/- 16% of baseline at 4 minutes, respectively. Recovery rates of post-transplant cardiac function in the load group were worse than in the unload group (LV Emax: 64 +/- 8 vs 84 +/- 5%, p < 0.05). Levels of post-transplant CK-MB in the load group were higher than in the unload group (639 +/- 119 vs 308 +/- 70 IU/liter, p < 0.05). CONCLUSIONS: Cardiac arrest with circulatory load by asphyxiation caused more myocardial damage than unloaded arrest. This difference between the modes of death should be considered when evaluating the DCD hearts for clinical application. PMID- 19285620 TI - Low complication rates with totally implantable access port use in epoprostenol treatment of pulmonary hypertension. AB - BACKGROUND: Among patients with advanced pulmonary arterial hypertension (PAH) who are receiving epoprostenol treatment, complications due to the delivery system are known to be a cause of serious morbidity and mortality. In this study, we aimed to outline the complications associated with the use of a totally implantable access port (TIAP) and their consequences in continuous intravenous epoprostenol treatment. METHODS: One hundred eleven pulmonary hypertension (PH) patients treated with epoprostenol through a totally implantable access port (TIAP) between May 1998 and July 2006 at three Dutch PH referral centers were retrospectively studied. RESULTS: During a mean follow-up period of 946 +/- 719 days, TIAP-related complications included local port site infections, bloodstream infections, port site skin perforations and incorrect port placement, with incidence rates of 0.11, 0.15, 0.06 and 0.04 per patient-year, respectively. Fatal, serious, moderate and minor consequences of these complications had incidence rates of 0.01, 0.50, 0.09 and 0.01 per patient-year, respectively. The median complication-free survival was 371 days. Staphylococcus aureus was the dominant infectious agent. The median TIAP life duration was 677 days. Female patients showed significantly longer first TIAP survival (p = 0.004). CONCLUSIONS: The use of a TIAP showed long-lasting port survival and relatively low complication rates. Our data suggest that the TIAP is well suited for continuous intravenous epoprostenol delivery in patients with pulmonary hypertension. PMID- 19285621 TI - Addition of prostanoids in pulmonary hypertension deteriorating on oral therapy. AB - BACKGROUND: The aim of our study was to describe the efficacy of addition of intravenous or subscutaneous prostanoids in idiopathic pulmonary arterial hypertension (PAH) patients deteriorating on bosentan or on bosentan-sildenafil. METHODS: PAH treatment at our hospital is standardized with first-line oral therapy in New York Heart Association class III patients followed by addition of prostanoids on clinical worsening. RESULTS: Mean improvement in 6-minute walk distance after 4 months of prostanoids was 86 m (p < 0.01) in the bosentan group versus 41 m (p < 0.05) in the bosentan-sildenafil group, and these improvements persisted at long-term follow-up. CONCLUSIONS: From these results we conclude that addition of subcutaneous or intravenous prostanoids can be efficacious in PAH deteriorating on oral therapy. PMID- 19285622 TI - Bone morphogenetic protein-2 stimulation of cartilage regeneration in canine tracheal graft. AB - BACKGROUND: Graft stenosis is among the most serious post-surgical complications that can occur after tracheal transplantation. Typically, stenosis is caused by resorption of tracheal cartilage. Bone morphogenetic protein-2 (BMP-2) is efficient at stimulating bone or cartilage regeneration. In this study, BMP-2 is tested for its effects on stimulation of cartilage regeneration in tracheal transplantation. METHODS: For tracheal autotransplantation, 24 mongrel dogs were divided equally into four groups and BMP-2 was injected between the cartilage rings at doses of 1, 3, 5 or 7 mg. For tracheal allotransplantation, 12 mongrel dogs were divided equally into two groups. One group received 5 mg of BMP-2 per graft, and the other received collagen only as a control. The grafts were harvested after 4 weeks and subjected to pathologic analysis. The diameter of the graft lumen and areas of new cartilage regeneration were measured. RESULTS: Regenerated cartilage areas were found in both the injected area and around the perichondrium. The areas of regenerated cartilage, as well as the diameter of the tracheal lumen, increased significantly with increasing concentrations of BMP-2. Five milligrams per milliliter was the most effective dose of BMP-2 in this study. CONCLUSIONS: BMP-2 can significantly stimulate cartilage regeneration in tracheal grafts and also can be used to prevent stenosis after tracheal transplantation. PMID- 19285623 TI - Cardiac recovery in a human non-heart-beating donor after extracorporeal perfusion: source for human heart donation? AB - Successful renal, liver and more recently lung transplantation using organs from non-heart-beating donors (NHBDs) has been reported. Regarding the heart, it has generally been assumed that warm ischemic insult would result in overwhelming and irreversible myocardial damage. We report recovery of cardiac function in a human NHBD by using extracorporeal perfusion 23 minutes after cardiorespiratory arrest. Successful cardiac resuscitation in the NHBD represents a potential source of increased donor organ supply for clinical heart transplantation. PMID- 19285624 TI - Recognition and management of complex rhythm disorders in heterotopic heart transplantation. AB - Managing arrhythmias is challenging in patients who have undergone heterotopic heart transplantation because of the superimposed rhythms of the native and donor hearts. We present the case of a 43-year-old man with a previously placed biventricular pacemaker who underwent heterotopic heart transplantation and later developed acute rejection of the donor heart, which led to bradycardia and pause dependent ventricular fibrillation. The patient remained clinically stable in the short term, likely because of partial recovery of myocardial function in the native heart. He later underwent placement of a pacing lead in the donor heart, allowing linking of the two hearts via a biventricular pacemaker. PMID- 19285625 TI - A pediatric case study of treprostinil overdose. AB - This case study reports the effects of a treprostinil accidental overdose in a patient with pulmonary hypertension. Treprostinil is labeled for treatment of pulmonary hypertension with New York Heart Association (NYHA) Class II, III and IV symptoms. It is characterized as a long-acting prostacyclin analog that can be delivered by intravenous or subcutaneous continuous infusions. This pediatric case report describes a massive overdose without significant side effects, thus suggesting a beneficial therapeutic index in pediatric patients. PMID- 19285626 TI - Nonabsorbable corticosteroids use in the treatment of gastrointestinal graft versus-host disease. AB - For over a decade, nonabsorbable corticosteroids have been employed in the treatment of gastrointestinal graft-versus-host-disease (GVHD) in hematopoietic stem cell transplant (HSCT), as monotherapy or in combination with systemic corticosteroids. The majority of the evidence showing a favorable outcome consisted of case series, small phase II trials and a large randomized phase III trial. The 2 most commonly studied molecules were oral budesonide and beclomethasone diproprionate. Although these reports hint at some benefit with the local treatment strategy, their methodologic inconsistencies preclude meaningful adoption to everyday clinical practice. This review evaluates the current evidence of nonabsorbable corticosteroids in HSCT and sets forth recommendations for future trials with these agents. PMID- 19285627 TI - Induction of graft-versus-leukemia (GVL) effect without graft-versus-host disease (GVHD) by pretransplant donor treatment with immunomodulators. AB - Pretransplant donor treatment with immunomodulators such as complete Freund's adjuvant (CFA) or oligodeoxynucleotide sequences expressing CpG motifs (CpG), was applied in sublethally irradiated host mice inoculated with murine models of mammary carcinoma (4T1) or B cell leukemia (BCL1). Spleen cells or IL-2 activated splenocytes (lymphokine activated killer [LAK]) derived from donor mice treated with CpG emulsified in incomplete Freund's adjuvant (IFA), (CpG + IFA) did not cause graft-versus-host disease (GVHD), but were not efficient enough to induce a significant graft-versus-tumor (GVT) response against 4T1 cells. In contrast, an efficient graft-versus-leukemia (GVL) effect was evident in BCL1-bearing mice inoculated with spleen cells from donors pretreated with CFA or CpG + IFA. Pretransplant donor treatment with CFA prolonged survival to a median of 62 days with 3 of 27 mice remaining GVHD- and leukemia-free for >200 days, compared to GVHD-related death of all mice inoculated with naive cells (median 17 days), or leukemia-related death of all mice inoculated with leukemia cells (median of 27 days). Pretransplant donor treatment with CpG + IFA exerted a more efficient GVL effect with reduced GVHD resulting in 12 of 26 GVHD- and leukemia-free survivors for >200 days. Our results suggest that it may be possible to prevent GVHD while sparing an efficient GVL effect by using pretransplant donor treatment with immunomodulators prior to allogeneic stem cell transplantation and/or donor lymphocyte infusions in hematologic malignancies. PMID- 19285628 TI - Frequency of abnormal findings detected by comprehensive clinical evaluation at 1 year after allogeneic hematopoietic cell transplantation. AB - Consensus guidelines recommend various screening examinations for survivors after allogeneic hematopoietic cell transplantation (HCT), but how often these examinations detect abnormal findings is unknown. We reviewed the medical records of 118 patients who received comprehensive, standardized evaluations at 1 year after allogeneic HCT at Fred Hutchinson Cancer Research Center/Seattle Cancer Care Alliance. Abnormal findings were common, including moderate to severe pulmonary dysfunction (16%), fasting hyperlipidemia (56%), osteopenia (52%), osteoporosis (6%), and active chronic graft-versus-host disease (cGVHD) (64%). Recurrent malignancy (4%) and cGVHD (29%) were detected in previously unsuspected cases. Only 3% of patients had no abnormal findings. We conclude that comprehensive evaluation at 1 year after allogeneic HCT detects a high prevalence of medical problems. Longer follow-up is needed to determine whether early detection and intervention affect later morbidity and mortality. PMID- 19285629 TI - Nonengraftment haploidentical cellular immunotherapy for refractory malignancies: tumor responses without chimerism. AB - Allogeneic bone marrow transplantation relies on immunosuppression, which controls graft-versus-host disease (GVHD) and allows engraftment at the expense of diminished graft versus-tumor (GVT) activity. Advances in hematologic transplantation have prompted the development of effective, less-toxic regimens that attempt to balance GVH and GVT immunoreactions. We analyzed the safety and efficacy of haploidentical transplantation in a Phase I/II nonimmunosuppressive, nonmyeloablative setting. A total of 41 patients with relapsed refractory cancer received 100 cGy of total body irradiation (TBI), along with an infusion of 1 x 10(6) to 2 x 10(8) CD3+ cells/kg; 29 patients received the highest dose. A postinfusional cellular graft rejection syndrome resembling engraftment syndrome was noted at the 2 highest CD3+ infusion cohorts. There were 26 patients with hematologic malignancies with 14 responses, 9 of which were major. Two of 6 patients with lymphoma remained free of disease at 76 months and 82 months, respectively; there were 5 durable complete responses and 4 partial responses in 13 patients with acute myelogenous leukemia (AML). All responses occurred outside of donor chimerism. TBI at 100 cGy followed by HLA-haploidentical immunotherapy is a biologically active therapy for patients with refractory AML and lymphoma. Possible mechanisms contributing to its effectiveness include initial GVT kill, breaking of host tolerance to tumor through cross-reactive alloreactive responses, persistent nondetectable microchimerism, or some combination of these. PMID- 19285630 TI - Allogeneic stem cell transplantation compared with chemotherapy for poor-risk Hodgkin lymphoma. AB - The aim of this study was to assess the role of allogeneic hematopoietic stem cell transplantation (HSCT) in patients with poor-risk Hodgkin's disease (HD) compared to chemotherapy. A donor was identified in 26 patients (14 HLA identical siblings and 10 alternative donors), and 24 received a transplant (Allo group). Twenty patients without a donor received different chemotherapy regimens and radiotherapy (CHEMO group). After a median follow-up of 28 months (range: 1-110), the 2-year overall survival (OS) was 71% in the ALLO group compared to 50% in the CHEMO group (P = .031). In the Allo group, the 2-year progression-free survival (PFS) was 47%. The 1-year nonrelapse mortality (NRM) in the ALLO group was 8% versus 0% in the CHEMO group. This study, suggests that allogeneic transplantation may prolong the survival in patients with a poor-risk HD. PMID- 19285631 TI - Incidence and risk factors of early bacterial infections after unrelated cord blood transplantation. AB - Incidence and characteristics of early bacterial infection within 100 days after unrelated cord blood transplantation (UCBT) were assessed for 664 pediatric and 1208 adult recipients in Japan. Cumulative incidence of early bacterial infection at day 100 post-UCBT was 11% (95% confidence interval [CI], 8%-13%) for children and 21% (CI, 19%-24%) for adults (P < .0001). Early bacterial infection in adults had a significant impact on mortality (hazard ratio [HR] = 2.1, CI, 1.7-2.6; P < .0001), although no significant risk factors were identified. Multivariate analysis identified older age group (6-10, and 11-15 years versus 0-5 years of age) at transplant (HR = 2.0 and 2.7, CI, 1.1-3.5 and 1.4-4.9; P = .020 and .002, respectively) as an independent risk factor of early bacterial infection for children. Early bacterial infection in children did not have a significant impact on mortality when adjusted. Of 315 bacteremia, 74% were caused by Gram-positive microorganisms. Pneumonia occurred in 39 patients including 13 cases of Stenotrophomonas maltophilia pneumonia. Early bacterial infection had a negative effect on survival for adults and the median day of development was 10 days after transplant, suggesting that the prevention of bacterial infection in the very early post-UCBT phase is important. PMID- 19285632 TI - Carbon monoxide diffusion capacity: how low can you go for hematopoietic cell transplantation eligibility? AB - Current guidelines suggest that patients with a pretransplantation diffusion capacity of the lung for carbon monoxide (DLCO) < or = 60% are not ideal candidates for hematopoietic cell transplantation (HCT); however, recent studies suggest this criterion may exclude patients who will benefit from the procedure. We conducted a study of all adult patients who underwent autologous or allogeneic HCT between 1990 and 2005, and had a DLCO < 60%, of predicted normal, to examine whether there is a lower limit for the DLCO threshold in the context of respiratory failure and nonrelapse mortality risk (NRM), and whether a comprehensive risk scoring system, such as the Pretransplant Assessment of Mortality (PAM) risk score, can more effectively risk stratify these patients with a very low pretransplantation DLCO. We found that among patients with a DLCO < or = 60% the risk of respiratory failure or NRM was not significantly different; however, the PAM score effectively risk-stratified these patients for NRM risk. There was a stepwise relationship between PAM score category and NRM risk; the highest PAM score category was associated with a 4.38-fold increase in risk (P < .001). These findings suggest that the pretransplantation DLCO should not be considered the sole eligibility criteria for allogeneic HCT. PMID- 19285633 TI - HLA-mismatched unrelated donors as an alternative graft source for allogeneic stem cell transplantation after antithymocyte globulin-containing conditioning regimen. AB - Between August 1996 and December 2004, 369 patients with a median age of 41 years (range: 1-68 years) received stem cell transplantation (SCT) from unrelated donors after an antithymocyte-globulin (ATG)-containing conditioning regimen. In 268 patients, complete molecular typing (4-digit) of HLA-A, -B, -C, -DRB1, and DQB1 was available: 110 patients were completely matched for 10 alleles, 91 patients had 1 allele-mismatch (9/10), and 67 patients were mismatched for 2-4 alleles (6-8/10). The incidence of grade II-IV acute graft-versus-host disease (aGVHD) was 33% in the 10/10, 41% in the 9/10, and 40% in the 6-8/10 group, respectively (P = .1). The cumulative incidence of treatment-related mortality (TRM) and relapse among the groups were similar (27%, 31%, and 32%, P = .2; and 28%, 27%, and 26%, P = .9. After a median follow-up of 35 months (range: 3-120 months), the estimated 5-year disease-free survival (DFS) was 42% and did not differ among the 10/10, the 9/10, and the 6-8/10-mismatched groups (45% versus 42% versus 39%) (P = .5). In multivariate analysis, only age (hazard ratio [HR] 1.013) (P = .004) and bad-risk disease (HR 1.975) (P < .001) were independent risk factors for DFS. In conclusion, pretransplant ATG allows allogeneic SCT from unrelated donors with HLA disparities. PMID- 19285634 TI - Complete remission status before autologous stem cell transplantation is an important prognostic factor in patients with multiple myeloma undergoing upfront single autologous transplantation. AB - Upfront high-dose myeloablative chemotherapy followed by a single autologous stem cell transplantation (ASCT) is the standard therapy for patients under the age of 65 years with newly diagnosed multiple myeloma (MM). Because disease status after induction chemotherapy is variable, we evaluated the prognostic effect of disease status before ASCT, especially in patients who were initially chemosensitive. We retrospectively analyzed the initially chemosensitive MM patients (> or = partial remission [PR]) enrolled in the Korean Multiple Myeloma Working Party Web-based registration system (www.myeloma.or.kr). Between November 1996 and January 2007, 197 MM patients (median age 53 years) were treated with induction chemotherapy followed by a single ASCT. All patients received peripheral blood stem cell (PBSC) support after conditioning with melphalan (Mel) alone. We considered those patients with no detectable M-protein regardless of the result of immunofixation to be in complete remission (CR) in this study. The median follow-up times were 29.2 months (range, 5.4 to 103.8 months) from the day of diagnosis and 22.4 months (range, 0.4 to 96.0 months) from the day of ASCT. Before ASCT, 63 patients (32%) were in CR and 134 (68%) were in partial remission (PR). The patients in CR had significantly longer overall survival (OS) from the day of ASCT compared with those in PR (P = .0015). Among the patients who received induction chemotherapy with vincristine, adriamycin, and dexamethasone (n = 162), the same difference in OS was seen between those in CR and those in PR before ASCT (P = .0016). CR after ASCT also predicted longer OS (P = .0135); however, patients with continued CR after ASCT had significantly higher OS after ASCT compared with patient with induced CR after ASCT who were in PR before ASCT (P = .0178). Multivariate analysis indicated that remission status pre-ASCT (CR vs PR) is a significant prognostic factor for predicting OS after ASCT (P = .012, Cox proportional hazard analysis; odds ratio = 2.83; 95% confidence interval = 1.25 to 6.37). We conclude that patients with MM who are in CR before ASCT have a better OS than those in PR before ASCT. Continued CR after ASCT may be an important prognostic factor as well. Our findings suggest that the development of more effective induction regimens, including novel antimyeloma agents to improve initial response, should be pursued to enhance clinical benefits post-ASCT. PMID- 19285635 TI - Peripheral blood eosinophilia has a favorable prognostic impact on transplant outcomes after allogeneic peripheral blood stem cell transplantation. AB - Peripheral eosinophilia after allogeneic stem cell transplant (ASCT) may reflect the activation of the Th2 cytokine pathway. A retrospective analysis was performed to evaluate the impact of early- (before day 100: EEo) or late-onset (beyond day 100: LEo) eosinophilia (> or =0.5 x 10(9)/L in peripheral blood) on transplant outcomes after peripheral blood SCT (PBSCT) in 237 patients. The incidence of EEo and LEo was 43% at day 100 and 62% at 2 years, respectively. Compared with patients without LEo, improved transplant outcomes were observed in patients with LEo: better overall survival (OS; 86% versus 41%, P = 5 x 10(-11)), lower nonrelapse mortality (NRM; 10% versus 37%, P = 3 x 10(-6)), lower relapse incidence (11% versus 31%, P = 3 x 10(-5)), and higher GVHD-specific survival (GSS; 90% versus 64%, P = 1 x 10(-6)) were observed. In addition, similar finding was observed when transplant outcomes were analyzed according to the occurrence of eosinophilia at the onset of cGVHD. The multivariate analyses confirmed a favorable implication of LEo on OS, NRM, and GSS. LEo was associated with: (1) less severe chronic GVHD (cGVHD), (2) higher prevalence of autoantibodies, and (3) rapid lymphocyte count recovery after ASCT. In summary, the development of eosinophila after allogeneic PBSCT seemed to be a prognostic marker for improving transplant outcome. PMID- 19285636 TI - A retrospective review of the outcome after second or subsequent allogeneic transplantation. AB - The failure of allogeneic stem cell transplant (allo-SCT) is cumbersome. We analyzed our experience in a second allo-SCT. Between the years 1981 and 2007, 144 patients underwent 2 or more allo-SCT. The first to second transplant interval ranged from 18 days to 13.25 years (median 98 days). The most frequent indications for the second SCT were activity of the basic disease (78), rejection (37), and engraftment failure (25). Twenty-nine of the 144 (20%) patients transplanted survived more then a year with treatment-related mortality of 45.5% as the leading cause of death. Interestingly, despite the low rate of graft versus-host disease (GVHD) prophylaxis used, only 51 and 16 of the patients developed acute and chronic GVHD (aGVHD, cGVHD), respectively. Factors indicating higher likelihood for survival were nonmalignant disease, a nonrelapse indication for the second SCT, full HLA-matching, and the use of reduced-intensity conditioning (RIC). Age at transplantation, time interval between transplants, the development of GVHD, conditioning regimen, GVHD prophylaxis, or graft source were not shown to influence the prognosis. With a median follow-up of 4.5 years, 25 patients (17.2%) are alive, and 18 are disease-free. We conclude that although toxic, a second allo-SCT can lead to long-term survival. PMID- 19285637 TI - Clinical utility of computed tomography screening of chest, abdomen, and sinuses before hematopoietic stem cell transplantation: the St. Jude experience. AB - All allogeneic (allo) and autologous (auto) hematopoietic stem cell transplantation (HSCT) recipients at St. Jude Children's Research Hospital undergo pre-HSCT computed tomography (CT) of the sinuses, chest, and abdomen because they are at significant risk for opportunistic infections. We studied whether this extensive routine imaging is warranted to detect infection despite the risk of additional radiation exposure. We reviewed the medical records of all children receiving allo- and auto-HSCT at St. Jude in 2004 and 2005. Of the 184 eligible patients who received 187 transplants, 131 received allografts and 56 autografts. Solid tumors and lymphomas were removed from the final analysis of the chest and abdomen CT as this imaging is typically warranted as part of disease restaging; thus, 111 allogeneic participants were included in this analysis. Both auto- and allo-recipients were evaluated by sinus CT and included in this final analysis. Most allo- and auto-HSCT recipients (> or =80%) did not have sinus, pulmonary, cardiac, or gastrointestinal symptoms; >85% of the evaluable allo-recipients had no prior fungal infection. Eighty-eight allo- and 31 auto-HSCT recipients had abnormal sinus CT findings, all unrelated to the underlying disease. Sixty-two (55.9%) of the allo-recipients had normal chest CT and 85 (76.6%) had normal abdominal CT. Of the 18 allo-recipients who began new therapy based on these findings, only 2 (11.1%) were related to chest CT findings and the other 16 were related to sinus findings. Our findings suggest that pre HSCT routine CT imaging of the abdomen may not be warranted in a subset of allogeneic recipients who are asymptomatic and without previous infectious findings. Thus, these patients may be spared unnecessary radiation exposure. Recipients undergoing auto-HSCT or allo-HSCT for lymphomas or solid tumors will routinely undergo chest and abdominal CT imaging as part of their disease evaluation. The decision to perform chest CT should be made judiciously based on a careful history and physical examination. Sinus imaging, which was frequently abnormal, may be justified in all patients to plan post-HSCT care. PMID- 19285638 TI - Immune reconstitution following myeloablative allogeneic hematopoietic stem cell transplantation: the impact of expanding CD28negative CD8+ T cells on relapse. AB - Allogeneic stem cell transplantation has become standard therapy for hematologic malignancies through the positive immunologic graft-versus-leukemia effect. Initial immune recovery relies on peripheral expansion of infused T cells, which switch to a memory-like phenotype. This study prospectively investigated whether changes in subset composition precedes complications after myeloablative HLA matched transplantation for hematologic malignancies. Of 80 allograft recipients, 18 were still free of clinical complication throughout 395 to 1564 days of follow up. Compared with this complication-free subgroup, patients who developed chronic graft-versus-host disease (cGVHD) without relapsing recovered similar numbers of circulating T cells with predominance of CD8+ T cells lacking CC-chemokine receptor-7 and CD28 expression throughout the first year after transplantation. Conversely, poor CD8+ T cell recovery with diminished numbers of CD28neg CD8+ T cells (approximately 1/4th of that of relapse-free patients) preceded occurrence of malignant relapse. In multivariate analysis, lower CD28neg CD8+ T cell counts by day 60 postallograft were associated with a greater risk of subsequent relapse (hazard ratio [HR] 0.33; 95% confidence interval [CI]: 0.14-0.76; P = .01). Enumeration of CD28neg CD8+ T cells in patients could assist in predicting risk of relapse and help build an algorithm for accelerating the immune recovery by reducing the immunosuppressive treatment and considering the introduction of preemptive donor lymphocyte infusions. PMID- 19285639 TI - First-line therapy for chronic graft-versus-host disease that includes low-dose methotrexate is associated with a high response rate. AB - We report the results of low-dose methotrexate (MTX) as first-line therapy mostly in combination with other immunosuppressive agents in patients with chronic graft versus-host disease (cGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Between November 2001 and March 2008, 86 patients with cGVHD after allo-HSCT received low-dose MTX therapy until a complete or partial response (CR, PR) was achieved, or until treatment failure or intolerable side effects were found. The median time from HSCT to the start of MTX was 154 (range: 80-993) days. The median number of MTX administrations was 4 (range: 2 18). The overall response rate among all enrolled patients was 83% (71 of 86 patients). The response rate for GVHD involving various organs was 90% (45 of 50) in the skin, 75% (39 of 52) in the liver, 42% (5 of 12) in the mouth, 3 of 7 in the eye, and 2 of 2 in the gut. In addition, MTX treatment allowed for a significant reduction in the prednisone dosage (median 90%) from 20 (2.5-100) mg at the start of MTX administration to 5 (0-30) mg 1 month after MTX was last used. Multivariate analysis showed that the only significant factor related to higher CR rate was sole organ involvement (P = .007). Grade 3 toxicities occurred in only 3 patients presenting cytopenias or oral mucositis. From this analysis, MTX appears to be a well-tolerated, effective, and inexpensive agent when used as a first-line treatment in combination with other immunosuppressive agents for cGVHD, especially for skin or sole organ involvement without concomitant thrombocytopenia. PMID- 19285640 TI - Imipenem/cilastatin with or without glycopeptide as initial antibiotic therapy for recipients of autologous stem cell transplantation: results of a Spanish multicenter study. AB - We analyzed the efficacy of imipenem/cilastatin alone (group I, 197 patients) or in combination with a glycopeptide (group I + G, 231 patients) as first-line antibiotic therapy for 2 consecutive cohorts of autologous stem cell transplantation (ASCT) recipients with febrile neutropenia. From June 2001 to June 2002, patients received imipenem/cilastatin (500 mg/6 hours), and from July 2002 to December 2003, they received imipenem/cilastatin as for group I plus a glycopeptide (vancomycin, 1 g/12 hours or teicoplanin, 400 mg/day). Fever of unknown origin accounted for 33.5% of episodes (66 patients) in group I and 50% of episodes (116 patients) in group I + G (P = .005). Bacteremia occurred in 55 patients (28%) in group I and in 51 patients (22%) in group I + G (P = .16). Resolution of fever without modification of the therapy regimen was observed in 108 patients (55%) and 159 patients (69%) in groups I and I + G, respectively (P = .003). The median interval to defervescence (4 days) and overall mortality were similar between groups. Inclusion of a glycopeptide in the initial antibiotic regimen for febrile neutropenia results in a higher success rate without modifying the regimen. However, glycopeptide inclusion does not improve the interval to defervescence or mortality rate. PMID- 19285641 TI - Does extra alemtuzumab remove the graft-versus-leukemia effect after allogeneic stem cell transplantation in chronic lymphocytic leukemia? PMID- 19285642 TI - Low-dose foscarnet preemptive therapy for cytomegalovirus viremia after haploidentical bone marrow transplantation. PMID- 19285644 TI - Universal screening for postpartum depression: an inquiry into provider attitudes and practice. AB - OBJECTIVE: This study evaluated the use of the Edinburgh Postnatal Depression Scale (EPDS) for detection of postpartum depression (PPD) in an academic medical center outpatient population and assessed knowledge and attitudes of obstetric providers regarding detection and treatment of PPD. STUDY DESIGN: A total of 512 charts were reviewed for demographic, medical, and psychiatric information. Also, a validated e-mail survey was sent to University of North Carolina obstetrical providers (n = 47). RESULTS: Our chart review revealed that providers documented the EPDS score in 39% of visits and counseled patients on their score and/or depression in 35% of visits. The survey results show that all respondents agree that they are responsible for screening for PPD, and 94% are confident in diagnosing PPD. CONCLUSION: The majority of obstetric providers are not documenting the EPDS in their postpartum assessment, yet they feel responsible for and confident in screening for postpartum depression. PMID- 19285645 TI - Inconclusive results from the study evaluating the use of blunt needles during obstetric laceration repair. PMID- 19285646 TI - Face and construct validity of virtual reality simulation of laparoscopic gynecologic surgery. AB - OBJECTIVE: The objective of the study was to validate virtual reality simulation in assessing laparoscopic skills in gynecology by establishing the extent of realism of the simulation to the actual task (face validity) and the degree to which the results of the test one uses reflects the subject tested (construct validity). STUDY DESIGN: Subjects (n = 56) were divided into 3 groups: novices (n = 15), intermediates (n = 20), and experts (n = 21). Participants completed 3 repetitions of a training program consisting of 4 basic skills and 3 gynecologic procedural simulations. The performance was compared between groups using a post hoc Student t test with the Bonferroni technique. Face validity was determined by using a questionnaire of 27 statements. RESULTS: Resulting from the questionnaire, the opinion about the realism and training capacities of the tasks was favorable among all groups. The degree of prior laparoscopic experience was reflected in the outcome performance parameters of the tasks. Experts achieved significant better scores on specific parameters. CONCLUSION: The results of this study indicate acceptance and thus face validity of the system among both reference (novice, intermediate) and expert group. There is a significant difference between subjects with different laparoscopic experience and thereby construct validity for the laparoscopic simulator could be established. PMID- 19285647 TI - Biomechanical properties of synthetic and biologic graft materials following long term implantation in the rabbit abdomen and vagina. AB - OBJECTIVE: We sought to evaluate the effects of anatomic location and ovariectomy on biomechanical properties of synthetic and biologic graft materials after long term implantation. STUDY DESIGN: A total of 35 rabbits underwent ovariectomy or sham laparotomy and were implanted with polypropylene (PP) mesh (n = 17) or cross linked porcine dermis (PS) (n = 18) in the vagina and abdomen. Grafts were harvested 9 months later and underwent mechanical properties testing. RESULTS: After implantation, PS was similar in strength (P = .52) but was twice as stiff as PP (P = .04) and had a maximal elongation only half that of PP (P < .001). Degradation of PS was associated with decreased ultimate tensile strength (P = .03) and elastic modulus (P = .046). Vaginal PP grafts shrunk more (P < .001) and were less stiff than abdominal PP grafts (P = .049) but were not different in strength (P = .19). Ovariectomy had no effect (P > .05). CONCLUSION: Cross-linked PS undergoes long-term degradation resulting in compromised biomechanical properties and thus is likely inferior to lightweight PP meshes for pelvic organ prolapse and incontinence procedures. PMID- 19285648 TI - Validation of serum biomarkers for detection of early-stage ovarian cancer. AB - OBJECTIVE: Ovarian cancer has the highest mortality of all the gynecologic malignancies with most patients diagnosed at late stages. Serum CA-125 is elevated in only half of patients with stages I-II. We identified 3 serum proteins (apolipoprotein A-1, transthyretin, and transferrin) for the detection of ovarian cancer and reported them combined with CA-125 to effectively detect early-stage mucinous tumors. The objectives of this study were to assess the effectiveness of the panel in detection of early-stage serous and endometrioid ovarian cancers. STUDY DESIGN: In all, 358 serum samples (control, benign adnexal masses, and early-stage and late-stage ovarian cancer) were obtained from the National Cancer Institute. The level of each marker was measured. Multiple logistic regression models were built to calculate sensitivity and specificity. RESULTS: When combined with CA-125, the panel detected early-stage cancer with a sensitivity of 96%. The highest sensitivity was seen for detection of endometrioid subtype of early-stage carcinomas (98%). CONCLUSION: A panel of 4 serum biomarkers effectively detected early-stage ovarian cancers with the highest reported overall sensitivity of 96%. Endometrioid tumors were detected at early stages with a sensitivity of 98%. Prospective clinical analysis of the panel is needed to validate it as an effective screening tool for early-stage ovarian cancer. PMID- 19285649 TI - Maternal serum ADAM12 (A disintegrin and metalloprotease) in chromosomally abnormal pregnancy at 11-13 weeks. AB - OBJECTIVE: The objective of this study was to investigate the potential value of ADAM12 (A disintegrin and metalloprotease) in first-trimester screening for trisomy 21 and other major chromosomal abnormalities. STUDY DESIGN: The concentration of ADAM12 was measured at 11-13 weeks in cases of trisomy 21 (n = 49), trisomy 18 (n = 28), trisomy 13 (n = 20), Turner syndrome (n = 29), triploidy (n = 10), and euploid pregnancies (n = 272). The levels of ADAM12, expressed as multiples of median (MoM), were compared in cases and controls and were assessed for association with free beta-human chorionic gonadotropin (hCG) and pregnancy-associated plasma protein A (PAPP-A). RESULTS: The median ADAM12 value in trisomy 21 (0.961 MoM) was not significantly different from the euploid fetuses (1.013 MoM), but in trisomy 18 (0.697 MoM), trisomy 13 (0.577 MoM), triploidy (0.426 MoM), and Turner syndrome (0.747 MoM), the levels were significantly lower. In both the euploid and aneuploid pregnancies, there was a significant association between ADAM12 and free beta-hCG and PAPP-A. CONCLUSION: Maternal serum ADAM12 concentration at 11-13 weeks of gestation is unlikely to be useful in first-trimester screening for chromosomal abnormalities because in trisomy 21 the levels are not significantly different from normal, and in the other chromosomal defects, there is a significant association between ADAM12 and the traditional biochemical markers of free beta-hCG and PAPP-A. PMID- 19285650 TI - Specific systemic antioxidant response to preeclampsia in late pregnancy: the study of intracellular glutathione peroxidases in maternal and fetal blood. AB - OBJECTIVE: The physiopathology of preeclampsia is still unclear, but an imbalance between reactive oxygen species (ROS) and antioxidants, also called oxidative stress, appears to be an important contributing factor. The ROS promote lipid oxidation and are known to induce stress proteins, such as hemeoxygenase 1 (HO-1) and heat-shock protein 70 (Hsp-70). We hypothesized that glutathione peroxidases (GPx), a major class of antioxidant enzymes that regulate cell homeostasis by neutralizing lipid peroxides, are altered in the blood of preeclamptic women and neonates (venous cord blood). METHODS: Thirty-one preeclamptic and 30 normotensive pregnancies were recruited. The blood was fractionated using a discontinuous gradient to separate the different cell types. The messenger ribonucleic acid (mRNA) expression of GPx-1 and -4, HO-1, and Hsp-70 were analyzed by quantitative reverse transcriptase-polymerase chain reaction. GPx-1 and -4 protein level in blood cells was also detected by Western blot. The experiments were analyzed using the Student t test. RESULTS: The HO-1 and Hsp-70 mRNA expression in whole blood was significantly higher in both fetal and maternal circulations (P < .05). We also discovered that GPx-4 mRNA was 1.6-fold higher in blood of women with preeclampsia than in control pregnancies (P = .04). The latter was associated with an increase of both GPx-1 and GPx-4 protein and mRNA levels in the lymphocyte/monocyte fraction of the blood. Significantly higher GPx-4 mRNA levels in the fetal circulation of the preeclamptic group than the control group were also detected (P < .001). CONCLUSION: These data indicate that preeclampsia is associated with a specific antioxidant response in both maternal and fetal circulations, likely in response to the deleterious oxidative stress observed in this syndrome. PMID- 19285651 TI - Differential expression profile of microRNAs in human placentas from preeclamptic pregnancies vs normal pregnancies. AB - OBJECTIVE: The purpose of this study was to perform a comprehensive analysis of the microRNA expression profile in placentas from preeclamptic pregnancies vs normal placentas. STUDY DESIGN: Placentas were obtained from patients with (1) mild preeclampsia (n = 8) and (2) severe preeclampsia (n = 15) and (3) in a normal control group (n = 11) with elective cesarean delivery. The microRNA expression profile was assessed by microRNA microarray and real-time reverse transcriptase-polymerase chain reaction analysis. RESULTS: Thirty-four microRNAs were expressed differentially in preeclamptic placentas, compared with normal placentas. Of these, 11 microRNAs were overexpressed, and 23 microRNAs were underexpressed in preeclamptic pregnancies. Notably, several microRNA clusters on human chromosome 19q13.42, 13q31.3, Xq26.2, Xq26.3, and 14q32.31 (a human imprinted region) were expressed differentially in preeclamptic placentas. These results were confirmed with the use of real-time polymerase chain reaction for selected microRNAs (miR-210, -152, -411, and so on). CONCLUSION: The results show that 34 microRNAs are deregulated in preeclamptic pregnancies, which suggests the involvement of these microRNAs in the pathogenesis of preeclampsia. PMID- 19285652 TI - Effect of Lactobacillus rhamnosus GR-1 supernatant and fetal sex on lipopolysaccharide-induced cytokine and prostaglandin-regulating enzymes in human placental trophoblast cells: implications for treatment of bacterial vaginosis and prevention of preterm labor. AB - OBJECTIVE: The objective of the study was to determine the effect of fetal sex on the output of cytokines and prostaglandin-regulating enzymes in lipopolysaccharide (LPS) and probiotic lactobacilli-treated placental trophoblast cells. STUDY DESIGN: We examined the effect of LPS and Lactobacillus rhamnosus GR 1 supernatant in placental trophoblast cells on tumor necrosis factor (TNF) alpha, interleukin (IL)-1beta, and IL-10 using enzyme-linked immunosorbent assay and on prostaglandin-endoperoxide synthase 2 (PTGS2), 15-hydroxy prostaglandin dehydrogenase (PGDH), and toll-like receptor-4 (TLR-4) using Western blotting. Comparisons were performed using one-way analysis of variance and Student t test. RESULTS: LPS increased the output of TNF-alpha, IL-10, and PTGS2 with a greater response in male placentae. L rhamnosus GR-1 supernatant inhibited the LPS stimulated TNF-alpha and increased IL-10. It also up-regulated expression of PGDH in female placentae and partially reduced the LPS-stimulated PTGS2 in male placentae. There was no change in IL-1beta. Expression of TLR-4 was greater in placentae of male fetuses. CONCLUSION: These findings suggest an underlying mechanism for the sex difference in the incidence of preterm birth and provide potential evidence for a therapeutic benefit of lactobacilli in reducing preterm labor. PMID- 19285653 TI - Persistence of male hematopoietic CD34+ cells in the circulation of women does not affect prenatal diagnostic techniques. AB - OBJECTIVE: We aimed to verify whether fetal microchimerism, because of persisting fetal hematopoietic CD34(+) cells from previous pregnancies, could interfere with the development of genetic tests based on using these cells, isolated from maternal blood for the diagnosis of fetal aneuploidies. STUDY DESIGN: CD34(+) cells, isolated from blood of parous women with at least 1 son and nulliparous women, were analyzed by using qualitative polymerase chain reaction (PCR), quantitative PCR, and fluorescence in situ hybridization (FISH) to establish whether these molecular techniques are concurrently capable of detecting circulating male DNA. RESULTS: By qualitative PCR, male DNA was found both in parous and nulliparous women, whereas by quantitative PCR and FISH analyses, no male DNA or male nuclei were revealed except in 1 cultured CD34(+) sample from a nulliparous woman. CONCLUSION: Fetal hematopoietic CD34(+) cells can be used in the noninvasive prenatal testing of fetal aneuploidies because the presence of fetal microchimerism does not affect fetal diagnosis in current pregnancies. PMID- 19285654 TI - Objective vs subjective vault measurement after myopic implantable collamer lens implantation. AB - PURPOSE: To evaluate the relationship between subjective measurements of vault and objective values measured with Visante optical coherence tomography (OCT) in eyes receiving an implantable contact lens (ICL) for myopia correction. DESIGN: Observational cross-sectional study. METHODS: SETTING: Fernandez-Vega Ophthalmological Institute, Oviedo, Spain. PATIENTS: Four hundred and fifty-two eyes from 246 patients were elected to be implanted with a Visian ICL V4 (STAAR Surgical Inc, Monrovia, California, USA). OBSERVATION PROCEDURES: Subjective and objective measurements of vault after implantation of ICL. MAIN OUTCOME MEASURES: Subjective vault classified in 5 levels assessed using an optical section during slit-lamp examination. Objective vault was measured with Visante OCT (Carl Zeiss Meditec Inc, Dublin, California, USA). RESULTS: Average values of objective and subjective vault were 414 +/- 228 microm and 2.1 +/- 1.0, respectively and both parameters were highly correlated (r = 0.82; P < .001). Differences in average objective vault were statistically significant among the 5 groups of subjective vault (P < .001). Subjective vault 0 corresponded to a mean OCT value of 62 +/- 49 microm with 99% confidence interval (CI) [38; 86] microm; eyes with vault 1 to 203 +/- 93 microm with 99% CI [176; 230] microm; eyes with vault 2 to 402 +/- 131 microm with 99% CI [378; 425] microm; eyes with vault 3 to 594 +/- 146 microm, 99% CI [554; 633] microm; and vault 4 to 794 +/- 182 microm with 99% CI [713; 875] microm. CONCLUSIONS: Subjective and objective values of vault are highly correlated. In 99% of cases within the CI, objective values for eyes subjectively classified within a certain level vary within a narrow interval (+/-25 to 80 microm) around the mean value, and this interval is characteristic of each subjective level. PMID- 19285655 TI - The occurrence of monofixational exotropia after exotropia surgery. AB - PURPOSE: To study patients with monofixation and presumed intermittent exotropia with monofixation with the specific intent of determining if they should be categorized as separate diagnostic entity than intermittent exotropes with bifoveal fusion. DESIGN: A retrospective re-analysis of data from 2 previously published prospective randomized clinical trials on the treatment of intermittent exotropia with attention to the occurrence of the monofixation syndrome. METHODS: A single-center institutional practice study of 215 patients from 2 separate consecutive randomized series undergoing surgery for presumed intermittent exotropia. Outcome measure was the presence of monofixation after surgery. RESULTS: Of 215 patients undergoing surgery for presumed intermittent exotropia, 194 were over 3 years of age at surgery, had bifoveal fusion, and did not have a preoperative manifest microtropia. None of them developed the monofixation syndrome after surgery. An additional 14 patients who had been previously excluded from those studies because they were too young for sensory testing were included in this study. Seven of them (50%) had the monofixation syndrome after surgery. A different 7 patients who had also been excluded from those prior studies because they had a constant microtropia prior to surgery which would build on alternate cover testing were also included in this study. All 7 had the monofixation syndrome after surgery. CONCLUSION: The presence of the monofixation syndrome after surgery for presumed intermittent exotropia most likely reflects the fact that it was present preoperatively. Many of these patients manifest a constant microtropia preoperatively and hence should not be called intermittent exotropes. The term monofixational exotropia is more appropriately descriptive. PMID- 19285656 TI - One-year prospective intrapatient comparison of aspherical and spherical intraocular lenses in patients with bilateral cataract. AB - PURPOSE: To conduct longitudinal, intrapatient comparisons of aspherical and spherical silicone intraocular lenses (IOL) of the same material and platform in patients undergoing bilateral cataract surgery. DESIGN: Prospective, randomized study. METHODS: Sixty-two eyes of 31 patients were randomized to receive a silicone aspherical IOL (Tecnis Z9000; AMO Inc, Santa Ana, California, USA) in 1 eye and a silicone spherical IOL (CeeOn 911A; AMO Inc) in the other eye. Best spectacle-corrected visual acuity (BSCVA); corneal and ocular wavefront aberrations; contrast sensitivity under photopic (180 lux), intermediate (75 lux), and scotopic (15 lux) illumination; amount of IOL decentration and tilt; and degree of posterior capsular opacification were measured at 1, 3, 6, and 12 months after surgery. All-distance visual acuity (VA) was measured 3 months after surgery. RESULTS: There were no significant differences between IOLs with regard to BSCVA, amount of IOL decentration and tilt, degree of posterior capsule opacification, and all-distance VA at any point after surgery. Regarding corneal wavefront aberrations, there was no difference in third- and fourth-order root mean square (RMS). In ocular wavefront aberrations, aspherical IOLs showed significantly lower fourth-order RMS (P < .001) than spherical IOLs throughout the study, but not in third-order RMS. Contrast sensitivity under photopic and mesopic conditions was not different between IOLs, but contrast sensitivity under scotopic conditions was significantly better with aspherical IOLs than with spherical IOLs (P < .01) at all measurement points. CONCLUSIONS: The silicone aspherical IOL (Tecnis Z9000; AMO Inc) significantly reduced ocular spherical aberration and improved scotopic contrast sensitivity, and these results were consistent through the 1-year follow-up. PMID- 19285657 TI - Successful treatment of Stevens-Johnson syndrome with steroid pulse therapy at disease onset. AB - PURPOSE: To evaluate the visual prognosis of patients with Stevens-Johnson syndrome (SJS) and its severe variant, toxic epidermal necrolysis (TEN), followed by general and topical high-dose corticosteroids administration from disease onset. DESIGN: Prospective, observational case series. METHODS: Between May 1, 2003 and June 30, 2005, we enrolled 5 patients with SJS or TEN with ocular complications at the acute stage. Intravenous pulse therapy with methylprednisolone (steroid pulse therapy; 500 or 1000 mg/day for 3 to 4 days) was initiated within 4 days from disease onset. Topically, 0.1% betamethasone was applied over 5 times daily for at least 2 weeks. Visual acuity (VA) and slit-lamp microscopic appearance 1 year from disease onset were evaluated. RESULTS: At the first examination, corneal or conjunctival epithelial defects and pseudomembranous conjunctivitis were present in all cases. Skin eruptions dramatically improved after steroid pulse therapy. Although ocular inflammation increased for several days, pseudomembranes disappeared and corneal and conjunctival epithelium regenerated within 6 weeks. At the chronic stage, all eyes had clear corneas with the palisades of Vogt (POV), implying the presence of corneal epithelial stem cells. Best-corrected VA was 20/20 or better in all eyes. Five eyes showed superficial punctate keratopathy. No eye had cicatricial changes except for 1 with slight fornix shortening. No significant adverse effects of steroid occurred during all clinical courses. CONCLUSIONS: Steroid pulse therapy at disease onset is of great therapeutic importance in preventing ocular complications. Topical betamethasone also shows great promise for preventing corneal epithelial stem cell loss in the limbal region and cicatricial changes. PMID- 19285658 TI - Inguinal hernia repair with porcine small intestine submucosa: 3-year follow-up results of a randomized controlled trial of Lichtenstein's repair with polypropylene mesh versus Surgisis Inguinal Hernia Matrix. AB - BACKGROUND: The aim of this study was to evaluate the safety and efficacy of Lichtenstein's hernioplasty using Surgisis Inguinal Hernia Matrix (SIHM; Cook, Bloomington, Indiana) compared with polypropylene (PP; Angiologica, Pavia, Italy). METHODS: This was a prospective, randomized, double-blind trial comparing Lichtenstein's inguinal hernioplasty using SIHM versus PP. RESULTS: Seventy male patients underwent Lichtenstein's hernioplasty (n = 35 in the SIHM group and n = 35 in the PP group). At 3 years after surgery, there were 2 deaths (5.7%) in the PP group and 1 death (2.9%) in the SIHM group (not significant [NS]). Although the study was underpowered to evaluate the recurrence rate, only 1 recurrence (2.9%) was seen in the PP group (NS). Although a significant decrease in postsurgical pain incidence was never observed among patients in the SIHM group, a significantly lower degree of pain was detected at rest and on coughing at 1, 3, and 6 months and on movement at 1, 3, and 6 months and 1, 2, and 3 years. A significant decrease in postsurgical incidence and degree of discomfort when coughing and moving were observed among patients in the SIHM group at 3 and 6 months and at 1, 2, and 3 years after surgery. COMMENTS: SIHM hernioplasty seems to be a safe and effective procedure. PMID- 19285659 TI - Efficient and systematic synthesis of a small glycoconjugate library having human complex type oligosaccharides. AB - To prepare a small library of homogeneous glycoconjugates with varying oligosaccharide structures, a combinatorial strategy was employed. The target glycopeptide was divided into two peptide segments (A and B) and both were prepared by solid phase peptide synthesis. These peptides, which can be coupled by native chemical ligation through an amide bond, were subsequently coupled to two kinds of human complex type oligosaccharides. This process systematically afforded the desired glycoconjugate library. PMID- 19285660 TI - A kinematic approach to the conceptual representations of this and that. AB - The Italian demonstrative pronouns questo/a ('this([mas/fem])') and quello/a ('that([mas/fem])') implicitly convey information about objects' distance with respect to the speaker. Our study investigated the referents of questo/a ('this([mas/fem])') and quello/a ('that([mas/fem])') by analysing their influence on reach-to-grasp actions towards objects located near to (12cm) or far from (30cm) the participant. Upon acoustic instruction containing one or the other pronoun, participants had to reach for and grasp a target object. Results showed an effect of pronoun on movement planning. When the object was in the far position, reaction times were faster for the congruent pronoun quello/a, whereas when it was in the near position they were faster for the congruent pronoun questo/a. Interactions between implied and real target distance suggest that the conceptual spaces of questo/a and quello/a do not map onto a strict distinction between peripersonal and extrapersonal space. PMID- 19285661 TI - An integrated network-based mechanistic model for tumor growth dynamics under drug administration. AB - Cancer chemotherapy complexity ranges from the routes that the drug must follow before reaching the tumor site (pharmacokinetics), to the drug effects on tumor depletion (pharmacodynamics). Previous researchers, in their majority, have focused either on the pharmacokinetics (PK) or on the pharmacodynamics (PD) aspects of chemotherapy. Moreover, models that account for the molecular mechanisms of cancer development have limited scope in addressing the protein signals involved in tumor progression. For instance, the recently developed models for the p53 network, for which a number of mutations have been reported, must be integrated for further understanding of the disease. Here, we propose an integrated model that is composed of a compartmental PK/PD representation for drug therapy that incorporates p53 and cell cycle regulation. In particular, the dynamics of p53 and its network components, such as Mdm2, pRb, cyclin-cdk's, are modeled under drug administration. The results show that the proposed model is a realistic representation of the physiological expectations in a multi-scale, integrative approach. PMID- 19285662 TI - Acrosome formation-associated factor is involved in fertilization. AB - OBJECTIVE: To investigate the effects of a novel acrosome formation-associated factor (Afaf) on fertilization by its regulation of acrosomal exocytosis and endosomal trafficking. DESIGN: Controlled laboratory study. SETTING: Institution affiliated state key laboratory. SUBJECTS: ICR mice. INTERVENTION(S): Sperm penetration assay and in vitro fertilization experiment were performed to study the effects of the Afaf antibody on acrosome reaction and fertilization. Acrosome exocytosis (AE) with streptolysin O (SLO) permeabilization was conducted to test the Afaf's action in calcium events. Colocalization and coimmunoprecipitation was done to determine the interaction between Afaf and SNAP25 (synaptosome-associated protein of 25,000 daltons). Transferrin (Tf) uptake assay was performed to demonstrate the impact of Afaf on endosomal pathway. RNAi was used to rescue the inhibition of Afaf on Tf uptake. MAIN OUTCOME MEASURE(S): Number of penetrated sperms, in vitro fertilization rate. Acrosomal exocytosis index, relative Tf fluorescence. RESULT(S): The Afaf antibodies were capable of significantly inhibiting sperm penetration of the eggs, therefore reducing the rate of in vitro fertilization. Acrosome formation-associated factor was involved in calcium triggered AE by acting upstream of the calcium efflux from the acrosome inside. Acrosome formation-associated factor might exert an interaction with SNAP25, which is a crucial component in both exocytosis and endosomal trafficking. Acrosome formation-associated factor was also involved in the endocytic pathway by down-regulating Tf endocytosis in the HeLa cells, and the miRNA-mediated RNAi could rescue this alternation induced by Afaf. CONCLUSION(S): Acrosome formation associated factor might play an important role in membrane trafficking during acrosome formation and participate in fertilization. PMID- 19285663 TI - The views of adult offspring of sperm donation: essential feedback for the development of ethical guidelines within the practice of assisted reproductive technology in the United States. AB - OBJECTIVE: To provide an in-depth analysis of offspring attitudes toward their means of conception and the practice of sperm donation in the United States. DESIGN: Survey-based study using a 46-item questionnaire. PATIENT(S): Eighty-five adult offspring of sperm donation. INTERVENTION(S): Participants were recruited through an Internet-based support group for adults conceived through sperm donation. Eighty-five of them completed the questionnaire provided through a link to another Internet site. MAIN OUTCOME MEASURE(S): Responses to a 46-item questionnaire. RESULT(S): A majority of offspring learned of their conception at age >18 years during a planned conversation; had no information about their donor; viewed their donor as their "biological father"; had searched for and wanted identifying information on their donor and half-siblings; and supported the provision of extensive nonidentifying information or identity release in the practice of sperm donation. Participant attitudes toward their means of conception were evenly distributed from "very good" to "very bad." Other descriptive information on participants contributed to an understanding of their attitudes. CONCLUSION(S): Participant ratings of their conception were evenly distributed from "very good" to "very bad." Most believed that identifying information should be provided to recipients and that they themselves would not participate in the practice of gamete donation. PMID- 19285664 TI - Prokineticin 1 mRNA expression in the endometrium of healthy women and in the eutopic endometrium of women with endometriosis. AB - OBJECTIVE: To examine prokineticin 1 (PROK1) mRNA expression in eutopic endometrial glands obtained from patients with or without endometriosis, to investigate the presence of additional endometrial abnormalities in women with endometriosis. DESIGN: Prospective laboratory study. SETTING: University hospital. PATIENTS: Twelve control women and 12 patients affected by endometriosis in the secretory phase of the menstrual cycle. INTERVENTION(S): Endometrial specimens were obtained from women affected (cases) or not (control group) by endometriosis. Endometrial glands were freshly isolated from endometrial biopsies. MAIN OUTCOME MEASURE(S): PROK1 mRNA expression levels by real-time polymerase chain reaction analysis. RESULTS: PROK1 mRNA was detectable in 4 of 12 (33%) samples obtained from women affected by endometriosis, whereas 10 of 12 (83%) samples obtained from normal women were positive for PROK1 detection by real-time polymerase chain reaction. Moreover, detectable PROK1 mRNA levels were 10 times lower in samples obtained from women with endometriosis than in samples obtained from control women. CONCLUSION(S): PROK1 is a newly discovered angiogenic factor implicated in the vascular function of peri implantation endometrium and early pregnancy. An altered expression of PROK1 could be one of the several biochemical abnormalities characterizing eutopic endometrium in endometriosis. PMID- 19285665 TI - Testis atrophy and reduced sperm motility in transgenic mice overexpressing c FLIP(L). AB - OBJECTIVE: To study the effect of c-FLIP overexpression in testicular germ cells. DESIGN: A novel transgenic mouse model overexpressing the apoptotic modulator c FLIP in the testis was generated. SETTING: Animal facility and university research laboratory. ANIMAL(S): Transgenic mice overexpressing the long isoform of c-FLIP (c-FLIP(L)) under the transcriptional control of a 400 bp long regulatory region of the Stra8 promoter. INTERVENTION(S): Spermatozoa motility and testis histological, immunohistochemical, and Western blot analyses were carried out in transgenic and control derived specimens. MAIN OUTCOME MEASURE(S): Testis morphology, sperm motility, and germ cell apoptosis were assayed. RESULTS: Stra8 promoter was found to activate the ectopic overexpression of c-FLIP(L) in round and elongated spermatids. As a consequence of such overexpression, a dramatic loss of germ cells was observed, resulting in testicular atrophy associated with reduced sperm motility. CONCLUSION(S): The data show that c FLIP(L) forced expression in haploid male germ cells has detrimental effects on spermatogenesis and sperm quality and reveal a possible mechanism underlying the onset of testicular atrophy. PMID- 19285666 TI - Laparoscopy versus minilaparotomy in women with symptomatic uterine myomas: short term and fertility results. AB - OBJECTIVE: To retrospectively compare the feasibility, safety, morbidity, and pregnancy outcome of laparoscopy (LPS) and minilaparotomy (LPT) in the treatment of symptomatic uterine myomas. DESIGN: Retrospective, nonrandomized study. SETTING: Advanced Gynecological Endoscopy Center, Malzoni Medical Center, Avellino, Italy. PATIENT(S): 680 nonconsecutive patients with symptomatic uterine myomas. INTERVENTION(S): 350 women underwent LPS, and 330 underwent LPT myomectomy. MAIN OUTCOME MEASURE(S): Operative time, blood loss, hospital stay, pregnancy rate, and spontaneous abortion rate. RESULT(S): The mean operative time was 63 +/- 21 minutes (95% CI, 48-143) in the LPS group and 57 +/- 23 minutes (95% CI, 38-121) in the LPT group. The mean length of hospital stay was statistically significantly greater in the LPT group (3.1 +/- 0.5; 95% CI, 1-5) than the LPS group (2.1 +/- 0.8; 95% CI, 1-4). The overall spontaneous pregnancy rate after myomectomy was 53%; the pregnancy rate after LPS myomectomy (56%) was not statistically significantly higher than the rate for LPT (50%). CONCLUSION(S): Laparoscopy showed a lower morbidity than reported for the open approach and was characterized by less blood loss and a shorter postoperative hospitalization with an higher pregnancy rate. The operating time was not much longer in the laparoscopic group, and the intraoperative and postoperative complications appeared acceptable and not more than what is traditionally expected with the open approach. PMID- 19285667 TI - The human ovarian follicular fluid level of interleukin-8 is associated with follicular size and patient age. AB - OBJECTIVE: To investigate the relationship between interleukin-8 (IL-8) in the human ovarian follicle and follicular size, patient age, and fertility factors in IVF cycles. DESIGN: Prospective study. SETTING: University hospital research laboratory and infertility clinic. PATIENT(S): Women undergoing IVF with oocyte retrieval. INTERVENTION(S): Follicular fluid (FF) aspiration, oocyte isolation, FF storage, and experimental studies. MAIN OUTCOME MEASURE(S): Quantization of IL 8 by ELISAs and protein microarray; high-performance liquid chromatography (HPLC) followed by ELISA and Western blotting to evaluate alpha(2)-macroglobulin (alpha(2)M) bound IL-8; association of IL -8 to follicular size, patient age, and IVF outcomes. RESULT(S): Samples of FF from 63 patients contained an average of 629.59 pg/mL of IL-8 with 50%-70% bound to alpha(2)M. Large follicles contained higher levels of IL-8 than small follicles (937.34 vs. 86.97 pg/mL). The IL-8 concentration in the large follicles of women of young age was higher than that of older reproductive age women (1,373.61 vs. 673.29 pg/mL). There were no statistically significant associations found between IL-8 concentration and other IVF cycle factors or pregnancy outcome. CONCLUSION(S): Our findings indicate that IL-8 is present in FF, both in its free and alpha(2)M-bound state, and its concentration is correlated with follicular size and patient age. PMID- 19285668 TI - Changes in acetylation on lysine 12 of histone H4 (acH4K12) of murine oocytes during maternal aging may affect fertilization and subsequent embryo development. AB - OBJECTIVE: To compare acH4K12 levels in oocytes during mouse aging and then assess how such changes might affect the developmental potential of oocytes. DESIGN: Experimental animal study. SETTING: State key laboratory and university research laboratory. ANIMAL(S): Kunming white strain mice. INTERVENTION(S): Oocytes obtained from TSA treated group or aging mouse group were fertilized and the formation of pronuclei and subsequently developmental potential in vitro or in vivo were assessed. MAIN OUTCOME MEASURE(S): AcH4K12 levels in oocytes were assessed using fluorescence staining, and confocal microscopy and oocyte developmental potentials were determined by in vitro or in vivo methods. RESULT(S): The AcH4K12 levels in oocytes statistically significantly increased during mouse aging. When histone acetylation of oocytes of young mice was artificially increased by trichostatin A (TSA) treatment, the acH4K12 levels in male and female pronuclei in fertilized oocytes showed statistically significant changes. About 38.9% of TSA-treated oocytes failed to form pronuclei or formed morphologically abnormal pronuclei 6 hours after fertilization, which statistically significantly decreased the blastocyst rate of TSA-treated oocytes when compared with the control group (41.5% vs. 60.5%). A similar reduction in blastocyst development was also observed when oocytes collected in older mice were compared with younger mice (17.3% vs. 69.4%). CONCLUSION(S): The AcH4K12 levels in oocytes statistically significantly increased during the aging process in mice, and such changes may affect the acetylation patterns and morphology of pronuclei during fertilization and lead to a reduction in oocyte developmental potential. PMID- 19285669 TI - Unusual complication of excision of pelvic endometriosis: pseudoaneurysm of the left uterine artery. AB - We report on a patient who had a pseudoaneurysm arising from the left uterine artery after surgical excision of deep endometriosis. The diagnosis was based on contrast-enhanced multidetector computed tomography angiography. Transfemoral selective catheterization and embolization of the left uterine artery determined a quick improvement of the symptoms. PMID- 19285670 TI - The condylar plate for treatment of distal femoral fractures: a long-term follow up study. AB - The aim of this retrospective study was to present the long-term functional and radiological outcomes of indirect reduction techniques and fixation with a condylar plate for treatment of distal femoral supracondylar or intracondylar femoral fractures. The series included 24 men and 17 women, mean age 51 years, between March 1994 and April 1999. All fractures were AO type 33, and eight were open fractures. Primary iliac bone graft was used in five cases. In one case of severe osteoporosis, screw fixation was augmented with cement. There were three delayed unions, one non-union and two infections; four participants required reoperation with bone grafts. Two (5%) participants developed a second varus deformity and three a second valgus deformity; correction osteotomy with bone grafts was necessary in these cases. After a mean follow-up of 9.5 years, the mean Neer score was 82 points and indicated that function was excellent in 16, satisfactory in 9, unsatisfactory in 4 and poor in 2 cases. The mean Neer score in cases of isolated fracture was 89 points and in cases with additional injuries was 72 points. Thus the long-term results of indirect reduction techniques of distal femoral fractures treated with the condylar plate were good to excellent in 82% of cases. PMID- 19285671 TI - Open acute segmental tibial fracture fixation using the Less Invasive Stabilisation System (LISS): study of 23 consecutive cases. AB - AIM: To evaluate the Less Invasive Stabilisation System (LISS) in the treatment of segmental tibial fractures. METHODS: A prospective series of 23 consecutive open acute segmental tibial fractures were treated with a long LISS plate, in a university hospital. All but three fractures were temporarily fixed with a unilateral external frame. The minimal follow-up was 2 years. RESULTS: Mean age of the casualties was 34 (17-72) years, all but two were male and all were involved in high-energy accidents, had multiple injuries and open fractures. Of the fractures, 4 were grade IIIA, 16 grade IIIB and 3 grade IIIC. It was possible to span the distal fracture with the LISS plate in 20 cases; of these, 3 developed compartment syndrome. Two people with trifocal fractures required additional distal plates, one of which became loose and infected, resulting in a malunion of 8 degrees. Dermatofasciotomy was performed in 13 cases, in 10 of these as a preventive measure and in 3 because of impending compartment syndrome. In five cases a post-fixation neural palsy of the extensor hallucis longus was observed. All except three fractures healed; mean time for union for the proximal fractures was 16 (10-24) weeks and for the distal fractures was 22 (16-44) weeks. CONCLUSION: Use of the LISS plate in fixing these difficult fractures is commended, but immediate fixation should be avoided. A staged sequential procedure with a temporary spanning external fixation and definitive treatment after soft-tissue healing is preferred. PMID- 19285672 TI - New surgical staging of endometrial cancer: 20 years later. PMID- 19285673 TI - Neuromuscular and balance responses to flywheel inertial versus weight training in older persons. PMID- 19285674 TI - Thermal desorption gas chromatography-mass spectrometry as an enhanced method for the quantification of polycyclic aromatic hydrocarbons from ambient air particulate matter. AB - Polycyclic aromatic hydrocarbons (PAHs) from ambient air particulate matter (PM) were analysed by a two-step thermal desorption (TD) injection system integrated to a gas chromatograph-mass spectrometer (GC/MS). The operational variables of the TD method were optimised and the analytical expanded uncertainties were calculated to vary from 8% to 16% over the operative concentration range (40-4000 pg). The performance of the TD method was validated by the analysis of a standard reference material and by comparison of PAH concentrations in PM samples to those obtained by a conventional liquid extraction (LE) method. The TD method reported lower uncertainties than the LE method for the analysis of similar concentrations in air. The TD method also showed advantages for shorter sampling times in comparison to 24 h for source apportionment applications and for reducing losses of more reactive compounds such as benzo[a]pyrene. PMID- 19285675 TI - Peptide mapping with mobile phases of intermediate pH value using capillary reversed-phase high-performance liquid chromatography/electrospray ionisation tandem mass spectrometry. AB - This investigation describes the separation of tryptic peptides by capillary reversed-phase high-performance liquid chromatography (RP-HPLC) with eluents in the intermediate pH range, followed by in-line electrospray ionisation tandem mass spectrometry (ESI-MS/MS) analysis. For these purposes, gradient elution procedures with an aqueous eluent containing 20 mM ammonium formate, and an increasing content of acetonitrile or methanol, were employed. Compared to the analysis of the same tryptic peptides under low-pH conditions with an ion-pairing reagent, the increase in the pH with the 20 mM ammonium formate mobile phase led to significant changes in both peptide retention to the reversed-phase column and the collision-induced dissociation at the MS/MS stage as a consequence of the changes in the physico-chemical properties of these peptides, such as their overall charge, polarity and relative hydrophobicity. Thus, improved selectivity for the peptide separation and favourable tandem mass spectrometry analysis could be obtained with eluents in this intermediate pH range. The number of tryptic peptides identified by the new approach for the proteins investigated were significantly higher than that obtained by the conventional low-pH methods. Moreover, analysis of protein digests at very low concentrations was also performed under both acidic and intermediate pH conditions and similar improvements in selectivity and MS/MS detection limits were observed, i.e. identification of more distinct peptides and higher sequence coverage of the protein was obtained when eluents of intermediate pH were employed. This study therefore highlights the potential of conducting peptide mapping in the intermediate pH range to achieve more reliable and sensitive protein identifications with capillary RP-HPLC-ESI-MS/MS. PMID- 19285676 TI - Novel coating for solid-phase microextraction: electropolymerization of a molecular receptor functionalized with 2,2'-bithiophene for the determination of environmental pollutants at trace levels. AB - A novel solid-phase microextraction coating based on the electropolymerization of a molecular receptor properly functionalized with 2,2'-bithiophene was developed for the determination of environmental pollutants in air and water samples. The 2,2'-bithiophene-based film was electrochemically deposed on the surface of a gold wire using cyclic voltammetry. The fibres showed a porous structure with an average thickness of 12+/-3 microm (n=5): a good intra-batch and inter-batch repeatability with RSD lower than 14% was also observed. The selective interactions via hydrogen bonding with organic molecules containing electronegative atoms was proved: LOD values in the low ng/l range allowed the determination of vinyl chloride and chlorobenzenes at trace levels in environmental matrices. PMID- 19285677 TI - Investigation of the validity of the kinetic plot method to predict the performance of coupled column systems operated at very high pressures under different thermal conditions. AB - The present study investigates how strong the kinetic plot method is influenced by the changes in plate height, retention factor and apparent column permeability that arise under conditions of very high pressure. More precisely, the study investigates how well a set of performance measurements conducted on a single short column can be used to predict the performance of a long sequence of coupled columns. This has been investigated for the two practically most relevant thermal conditions, i.e., that of a forced-air oven and that of a still-air oven. Measuring column performance data for acetophenone and benzene on a series of coupled 3.5 microm columns that could be operated up to 1000 bar, it was found that the kinetic plot method provides accurate predictions of time versus efficiency for the still-air oven systems, over the entire range of investigated pressures and column lengths (up to 60 cm), provided k' and K(v0) are evaluated at the maximal pressure. For the forced-air oven which leads to worse performances than the still-air oven, the kinetic plot prediction is less accurate, partly because the thermal conditions (near-isothermal) tend to vary if the number of coupled columns increases. The fact that the thermal conditions of the column wall might vary with the column length is an additional complexity making very-high pressure separations less predictable and harder to interpret and model. PMID- 19285678 TI - Bacteriophage and impurity carryover and total organic carbon release during extended protein A chromatography. AB - In the biopharmaceutical industry, column chromatography residuals are routinely assessed by the direct measurement of mock eluates. In this study, we evaluated virus and other impurity carryover between protein A cycles and the feasibility of using a total organic carbon (TOC) analyzer to monitor for column impurity leakage as a correlate for actual measured carryover in mock eluates. Commercial process intermediates were used in scaled down studies of two protein A media, ProSep A (Millipore, Bedford, MA, USA) and MabSelect SuRe (GE Healthcare, Uppsala, Sweden). The chromatography system was programmed to run up to 200 normal load/elution cycles with periodic blank cycles to measure protein and phage carryover, and water flush cycles to measure TOC release. Sustained phage carryover was evident in each study. Carryover and TOC release was lowest in the case where cleaning was most stringent (50 mM NaOH/0.5 M Na(2)SO(4) with MabSelect SuRe). The TOC analysis at this time does not appear to be a viable practical means of measuring impurity carryover; direct measurements in mock eluates appears to be more predictive of column performance. PMID- 19285679 TI - Optimizing the peak capacity per unit time in one-dimensional and off-line two dimensional liquid chromatography for the separation of complex peptide samples. AB - To obtain the best compromise between peak capacity and analysis time in one dimensional and two-dimensional (2D) liquid chromatography (LC), column technology and operating conditions were optimized. The effects of gradient time, flow rate, column temperature, and column length were investigated in one dimensional reversed-phase (RP) gradient nano-LC, with the aim of maximizing the peak per unit time for peptide separations. An off-line two-dimensional LC approach was developed using a micro-fractionation option of the autosampler, which allowed automatic fractionation of peptides after a first-dimension ion exchange separation and re-injection of the fractions onto a second-dimension RP nano-LC column. Under the applied conditions, which included a preconcentration/desalting time of 5 min, and a column equilibration time of 12.5 min, the highest peak capacity per unit time in the 2D-LC mode was obtained when applying a short (10 min) first-dimension gradient and second-dimension RP gradients of 20 min duration. For separations requiring a maximum peak capacity of 375, one-dimensional LC was found to be superior to the off-line strong cation exchange/x/RPLC approach in terms of analysis time. Although a peak capacity of 450 could be obtained in one-dimensional LC when applying 120-min gradients on 500-mm long columns packed with 3-mum particles, for separations requiring a peak capacity higher than 375 2D-LC experiments provide a higher peak capacity per unit time. Finally, the potential of off-line 2D-LC coupled to tandem mass spectrometry detection is demonstrated with the analysis of a tryptic digest of a mixture of nine proteins and an Escherichia coli digest. PMID- 19285680 TI - Fabrication of polyoxometalate-based nano/micrometer composite films by electrophoretic deposition method. AB - Keggin-type heteropolyanion H(3)PMo(12)O(40) encapsulated by a cationic surfactant dioctadecyldimethylammonium chloride (DODACl) has been assembled on ITO substrates using an electrophoretic approach. The films were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), IR spectra, X ray photoelectron spectra (XPS) and cyclic voltammetry. The AFM images exhibit a spherical assembly of surfactant-encapsulated complex (SEC) nanoparticles with uniform size. The SEM was also used to investigate the surface topography. It is the first report that the thin films of SEC are fabricated using this method, which provides a new route to explore the possibility of application to polyoxometalate-based hybrid inorganic-organic materials. PMID- 19285681 TI - Improving the diagnosis of bundle branch block by analysis of body surface potential maps. AB - Bundle branch block (BBB) is a defect on the electrical conduction system of the heart diagnosed by analyzing electrocardiogram (ECG) morphology. Our study aims to determine whether mapping information, specifically QRS duration and observation data from maps obtained using body surface potential mapping (BSPM), can be helpful in BBB diagnosis. We studied 64-lead BSPM recordings of 18 BBB patients and 9 controls with normal ventricular conduction. QRS duration was measured from the BSPM information obtained. The BSPM maps were computed along the QRS complex for each individual and group, and maps for each group were compared with maps for each individual. QRS complexes of the 12 standard leads were computed for each individual and group, and complexes of each group were compared with the complexes of each individual. QRS duration measured for all available leads (64 unipolar leads and 12 standard leads) was 7.4 +/- 3.9 milliseconds longer than QRS duration measured only in the standard 12-lead ECG for left BBB patients (LBBB) and 15.3 +/- 10.8 milliseconds longer for right BBB with left anterior fascicular block patients (RBBB_LAFB). In case of comparisons based on the standard ECG, sensitivity was 76.9% for LBBB patients and 66.6% for control subjects. However, classification based on map comparisons showed a sensitivity of 93% for LBBB patients and 89% for controls. QRS duration measured from BSPM information does not differ significantly from 12-lead standard ECG measurement for LBBB. However, differences are higher for RBBB_LAFB patients. Representative BSPM maps permit an automatic classification of the subjects. PMID- 19285682 TI - The predictive value of numerical magnitude comparison for individual differences in mathematics achievement. AB - Although it has been proposed that the ability to compare numerical magnitudes is related to mathematics achievement, it is not clear whether this ability predicts individual differences in later mathematics achievement. The current study addressed this question in typically developing children by means of a longitudinal design that examined the relationship between a number comparison task assessed at the start of formal schooling (mean age=6 years 4 months) and a general mathematics achievement test administered 1 year later. Our findings provide longitudinal evidence that the size of the individual's distance effect, calculated on the basis of reaction times, was predictively related to mathematics achievement. Regression analyses showed that this association was independent of age, intellectual ability, and speed of number identification. PMID- 19285683 TI - Operational momentum in large-number addition and subtraction by 9-month-olds. AB - Recent studies on nonsymbolic arithmetic have illustrated that under conditions that prevent exact calculation, adults display a systematic tendency to overestimate the answers to addition problems and underestimate the answers to subtraction problems. It has been suggested that this operational momentum results from exposure to a culture-specific practice of representing numbers spatially; alternatively, the mind may represent numbers in spatial terms from early in development. In the current study, we asked whether operational momentum is present during infancy, prior to exposure to culture-specific representations of numbers. Infants (9-month-olds) were shown videos of events involving the addition or subtraction of objects with three different types of outcomes: numerically correct, too large, and too small. Infants looked significantly longer only at those incorrect outcomes that violated the momentum of the arithmetic operation (i.e., at too-large outcomes in subtraction events and too small outcomes in addition events). The presence of operational momentum during infancy indicates developmental continuity in the underlying mechanisms used when operating over numerical representations. PMID- 19285684 TI - Children's representation of symbolic magnitude: the development of the priming distance effect. AB - The comparison distance effect (CDE), whereby discriminating between two numbers that are far apart is easier than discriminating between two numbers that are close, has been considered as an important indicator of how people represent magnitudes internally. However, the underlying mechanism of this CDE is still unclear. We tried to shed further light on how people represent magnitudes by using priming. Adults have been shown to exhibit a priming distance effect (PDE), whereby numbers are processed faster when they are preceded by a close number than when they are preceded by a more distant number. Surprisingly, there are no studies available that have investigated this effect in children. The current study examined this effect in typically developing first, third, and fifth graders and in adults. Our findings revealed that the PDE already occurs in first graders and remains stable across development. This study also documents the usefulness of number priming in children, making it an interesting tool for future research. PMID- 19285685 TI - Preschoolers' nonsymbolic arithmetic with large sets: is addition more accurate than subtraction? AB - Adult and developing humans share with other animals analog magnitude representations of number that support nonsymbolic arithmetic with large sets. This experiment tested the hypothesis that such representations may be more accurate for addition than for subtraction in children as young as 3(1/2)years of age. In these tasks, the experimenter hid two equal sets of cookies, visibly added to or subtracted from the sets, and then asked 3(1/2)-year-olds which set had more cookies. Initial set size was either large (7 or 9) or very large (18 or 30), and the final sets differed by either a high proportion (ratio of 1:2) or a low proportion (difference of 1 cookie). Children's addition performance exceeded chance, as well as their subtraction performance, across set sizes and proportions, whereas subtraction performance did not exceed chance. Arithmetic performance was also independent of counting ability. Addition performance was remarkably accurate when ratios between outcomes were close to 1, in contrast to previous findings. Interpretations for the asymmetry between addition and subtraction are discussed with respect to the nature of representations for nonsymbolic arithmetic with large sets. PMID- 19285686 TI - Erythropoietin attenuates lung injury in lipopolysaccharide treated rats. AB - BACKGROUND: Erythropoietin (EPO) elicits protective effects in lung ischemia reperfusion, hyperoxia, acute necrotizing pancreatitis, and some other tissues. In the present study, we investigated the possible protective roles of EPO in the lipopolysaccharide (LPS) induced lung injury. MATERIALS AND METHODS: Male Sprague Dawley rats were treated with EPO (3000 U/kg, i.p.) or vehicle (saline), 30 min prior to LPS administration (6 mg/kg, i.v.). Four h following LPS injection, samples of pulmonary tissue were collected. Optical microscopy was performed to examine pathological changes in lungs. Validated methods were used to measure wet/dry ratios (W/D), myeloperoxidase (MPO) activity, malondialdehyde (MDA) concentrations, and nitrite/nitrate (NO(2)(-)/NO(3)(-)) levels in lungs. Western blotting was performed to study the pulmonary expression of inducible nitric oxide synthase (iNOS) and nitrotyrosine protein. RESULTS: Pretreatment with EPO led to (1) significant attenuation of endotoxemia induced evident lung histologic injury and edema; (2) inhibition of LPS mediated induction in MPO activity and MDA concentration; (3) inhibition of LPS mediated overproduction of pulmonary NO(2)(-)/NO(3)(-) levels; and (4) marked suppression in endotoxin induced expression of iNOS and nitrotyrosine. CONCLUSIONS: This study provides considerable evidence that EPO has an ability to significantly attenuate endotoxin-induced acute lung injury in rats. PMID- 19285687 TI - New concepts for basal cell carcinoma. Demographic, clinical, histological risk factors, and biomarkers. A systematic review of evidence regarding risk for tumor development, susceptibility for second primary and recurrence. AB - Basal cell carcinoma (BCC) is the commonest cancer in Caucasians and its incidence is increasing. Whilst ultraviolet radiation (UVR) is recognized as the main etiological factor, the relationship between exposure and host phenotype is still unclear. We systematically searched Medline, Embase, and the Cochrane databases for studies assessing the genetic basis of host response to UVR DNA damage, the effect of UVR on generation of reactive oxygen species (ROS), and their detoxification, UVR induced skin immunity modifications, and the role of genomic instability with a focus on the potential use of these biomarkers to the surgical treatment planning and prognosis of BCC patients. Data suggest that risk for BCC development is likely to result from the combined effect of many genes, each with a relatively weak individual contribution. Certain genomic alterations have been associated with increased or reduced risk for BCC development, with a second primary BCC or with recurrence of BCC. However, use of these biomarkers in everyday practice should be supported by further studies, mainly for its cost effectiveness. In addition, not enough information exists on the prognostic value of existing demographic and clinical risk predictors for BCC regarding development of second primary or recurrent tumors. Information reviewed suggests that these predictors are of higher predictive value compared with biomarkers whilst they are indisputably cheaper and easier to monitor even in developing countries. Conclusively, we suggest that further studies aimed in investigating second primary or recurrent BCC are needed to provide better information on the predictive value of certain demographic, clinical and histological factors. PMID- 19285688 TI - Primary solid tumors of the colon and rectum in the pediatric patient: a review of 270 cases. AB - OBJECTIVE: To study the outcomes of solid tumors of the colon and rectum in pediatric patients. METHODS: The Surveillance, Epidemiology, and End Results (SEER) database (1973-2005) was queried for patients less than 20 y of age. RESULTS: Overall, 270 patients with malignant tumors of the lower gastrointestinal tract were identified. The annual incidence was 1.04 cases per million in 2005. Overall, equal distribution between boys (50.7%) and girls (49.3%) was observed. The majority of tumors arose in adolescents (68.1% were older than 15 y). Tumors were more commonly seen in white (77.8%) and non Hispanic (78.9%) patients. Tumors were identified in the right colon (45.9%), transverse colon (9.3%), left colon (20.4%), rectum (15.2%), and anal canal (1.1%). The most common histology of these tumors was adenocarcinoma (35.6%), followed by carcinoid (34.1%). Disease specific 5- and 10-y-survival for the entire cohort was 61.0% and 57.9%, respectively. Multivariate analysis of the cohort identified tumor stage (HR 8.39, P < 0.001 for distant disease), tumor type (signet ring HR 2.12, P = 0.025, and carcinoid HR = 0.14, P = 0.001), and surgical resection (no surgery HR 2.98, P = 0.010) as independent predictors of worse outcome. CONCLUSION: In the pediatric population, solid tumors of the colon and rectum occur more frequently in the right side of the colon in teenagers. Surgical resection significantly improves survival for most pediatric tumors of the lower gastrointestinal tract. PMID- 19285689 TI - Regional blood flow distribution and oxygen metabolism during mesenteric ischemia and congestion. AB - BACKGROUND: Acute mesenteric ischemia is a potentially fatal vascular emergency with mortality rates ranging between 60% and 80%. Several studies have extensively examined the hemodynamic and metabolic effects of superior mesenteric artery occlusion. On the other hand, the cardiocirculatory derangement and the tissue damage induced by intestinal outflow obstruction have not been investigated systematically. For these reasons we decided to assess the initial impact of venous mesenteric occlusion on intestinal blood flow distribution, and correlate these findings with other systemic and regional perfusion markers. METHODS: Fourteen mongrel dogs were subjected to 45 min of superior mesenteric artery (SMAO) or vein occlusion (SMVO), and observed for 120 min after reperfusion. Systemic hemodynamics were evaluated using Swan-Ganz and arterial catheters. Regional blood flow (ultrasonic flow probes), intestinal O(2)-derived variables, and mesenteric-arterial and tonometric-arterial pCO(2) gradients (D(mv a)pCO(2) and D(t-a)pCO(2)) were also calculated. RESULTS: SMVO was associated with hypotension and low cardiac output. A significant increase in the regional pCO(2) gradients was also observed in both groups during the ischemic period. After reperfusion, a progressive reduction in D(mv-a)pCO(2) occurred in the SMVO group; however, no improvement in D(t-a)pCO(2) was observed. The histopathologic injury scores were 2.7 +/- 0.5 and 4.8 +/- 0.2 for SMAO and SMVO, respectively. CONCLUSIONS: SMV occlusion promoted early and significant hemodynamic and metabolic derangement at systemic and regional levels. Additionally, systemic pCO(2) gradient is not a reliable parameter to evaluate the local intestinal oxygenation. Finally, the D(t-a)pCO(2) correlates with histologic changes during intestinal congestion or ischemia. However, minor histologic changes cannot be detected using this methodology. PMID- 19285690 TI - Comment on "Surgical resident performance on a virtual reality simulator correlates with operating room performance". PMID- 19285691 TI - A case of adult onset type II citrullinemia with portal-systemic shunt. AB - A 48-year-old woman who had conscious disturbance and abnormal behaviors had been misdiagnosed as having hepatic encephalopathy due to hyperammonemia and portal systemic shunt, and retrograde transvenous obliteration of the shunt did not improve her symptoms. Thereafter, analyses of plasma amino acids and citrin gene revealed a diagnosis of adult onset type II citrullinemia (CTLN2). She underwent auxiliary partial orthotopic liver transplantation (APOLT) using a left lobe graft from her brother, and her symptoms as well as hyperammonemia improved. Our case demonstrates the importance of CTLN2 as a differential diagnosis in patients with hyperammonemia and consciousness disturbance, even if they present with a portal-systemic shunt. PMID- 19285692 TI - Homocysteine and lipoprotein (a) correlation in ischemic stroke patients. AB - BACKGROUND: Homocysteine and Lipoprotein (a) have been recognized as risk factors for coronary heart disease. However, their role in ischemic stroke is still not defined. Therefore the present study was undertaken to evaluate their levels and relationship in patients of ischemic stroke. METHODS: The study was conducted in consecutive patients admitted with a diagnosis of acute ischaemic stroke and age and sex matched healthy controls. Plasma homocysteine and serum l lipoprotein (a) levels were determined in the fasting venous blood samples using ELISA and immunoturbidimetric assay respectively. RESULTS: 66 patients with ischaemic stroke (30 males, 36 females) of mean age 54.43+/-1.97 years and 72 controls (39 males, 33 females) of mean age 53.86+/-1.88 years were studied. Mean plasma homocysteine levels in the stroke patients and control groups were 28.40+/-2.08 micromol/L and 11.16+/-1.09 micromol/L respectively (p<0.001). Odds ratio for raised homocysteine levels in stroke cases was 15.7. Plasma homocysteine levels showed a positive correlation with smoking (Pearson's correlation coefficient=0.324 and p-value of 0.008), (Odds ratio=5.71). Serum Lipoprotein (a) levels in stroke cases and control group were 57.33+/-4.40 mg/dl and 23.46+/-1.09 mg/dl respectively, (p<0.001), (Odds Ratio=8.62). A positive correlation was also observed between Homocysteine and Lipoprotein (a) levels with Pearson's correlation coefficient of 0.75 and p-value<0.001. CONCLUSIONS: Raised homocysteine and serum lipoprotein (a) levels were found to be independently associated with ischemic stroke with a significant positive correlation between the two parameters. Elevated homocysteine levels may modulate the toxicity of lipoprotein (a) in ischemic stroke. PMID- 19285693 TI - The metabolic and fitness costs associated with metal resistance in Nereis diversicolor. AB - The population of Nereis diversicolor inhabiting the upper reaches of Restronguet Creek, Cornwall, UK is highly resistant to acute zinc and copper toxicity. Here we employ bioenergetic accounting and fecundity counts to demonstrate the energetic costs associated with this phenomenon in terms of the worms' allocation of metabolic resources and reproductive output [P(r)]. Metal-resistant animals exhibited a scope for growth that was 46-62% less than that of animals from two non-resistant reference populations, corresponding to a mean metabolic cost of 1.31 mJ h(-1)mg DW(-1). The resistant population also contained 13% less lipid than animals from the reference populations and 73-81% less carbohydrates. Consequently, mass-specific fecundity was reduced in resistant animals by 39-45%, although material investment in individual gametes did not appear to vary. This demonstrates fitness costs associated with metal resistance in this ecologically important polychaete and adds to our understanding of phenotypic trade-offs associated with resistance. PMID- 19285694 TI - Do 2006/7/EC European Union Bathing Water Standards exclude the risk of contact with Salmonella or Candida albicans? AB - The purpose of this study was to evaluate the efficiency of the recent (2006) European Union Directive concerning the management of bathing water quality to exclude the presence of pathogens from complying waters. Coastal water samples were classified according to the Directive. 2.5% of 'excellent' water contained Salmonella and 39.2% Candida albicans 11.8% of samples in category 'good' were Salmonella positive and 35.2% were C.albicans positive. When the USEPA criterion for marine waters was applied to the same dataset, fewer samples complied, however 6.0% of the complying samples contained Salmonella. The results suggest that the bacterial indicator threshold levels for marine recreational water quality of the 2006/7/EC European Union Directive do not fully exclude contact of bathers with dangerous pathogens. Enterococci, if used as the sole index, appear to form a more reliable proxy of the risk of contact with Salmonella. PMID- 19285695 TI - [Influence of cognitive reserve on neuropsychological performance in epileptic patients]. AB - BACKGROUND AND OBJECTIVE: Our goal was to determine the influence of the cognitive reserve (CR) on the neuropsychological performance of patients with temporal lobe epilepsy (TLE). PATIENTS AND METHOD: Twenty-eight patients with TLE from a program of epilepsy surgery were assessed with a neuropsychological battery that included standard clinical measures of memory, attention, visual construction skills and language. Raw scores adjusted according normative data were transformed to z scores. CR scores were based on a combination of educational level, occupational attainment and estimated premorbid intelligence. RESULTS: There were no significance differences in socio-demographic and clinical features between the 2 groups. Epileptic patients with low CR showed significantly lower scores than patients with high CR, with regard to measures of verbal memory (p<0.039), attention (p<0.001), visual-construction skills (p<0.019), and phonetic (p<0.003) and semantic (p<0.005) fluency. CONCLUSIONS: Epileptic patients with low CR showed greater neuropsychological morbidity than patients with high CR. These findings suggest that higher CR may decrease vulnerability or delay the clinical manifestation related to cognitive deficits following TLE. PMID- 19285696 TI - [The stroke programme of Catalonia]. PMID- 19285697 TI - Web-based resources for mass-spectrometry-based metabolomics: a user's guide. AB - In recent years, a plethora of web-based tools aimed at supporting mass spectrometry-based metabolite profiling and metabolomics applications have appeared. Given the huge hurdles presented by the chemical diversity and dynamic range of the metabolites present in the plant kingdom, profiling the levels of a broad range of metabolites is highly challenging. Given the scale and costs involved in defining the plant metabolome, it is imperative that data are effectively shared between laboratories pursuing this goal. However, ensuring accurate comparison of samples run on the same machine within the same laboratory, let alone cross-machine and cross-laboratory comparisons, requires both careful experimentation and data interpretation. In this review, we present an overview of currently available software that aids either in peak identification or in the related field of peak alignment as well as those with utility in defining structural information of compounds and metabolic pathways. PMID- 19285698 TI - [Idiopathic generalised epilepsies in the elderly: the viewpoint of a geriatrician]. AB - INTRODUCTION: Long-term follow-up studies indicate a low remission rate in idiopathic generalised epilepsies (IGE) (Martinez-Juarez et al., 2006), suggesting they may persist to an advanced age. However there are few estimates of IGE frequency in the elderly. METHODS: EEGs of 700 patients aged over 70 years, recorded between January 2006 and March 2007, were reviewed for anomalies consistent with IGE. We then examined the clinical history of patients with these anomalies. RESULTS: A persistent IGE was identified in four female patients (mean age: 79 years); in two cases it was a juvenile myoclonic epilepsy (JME) and in two an epilepsy with grand mal seizures. Seizures in three patients had begun in childhood or adolescence and in one at 40 years. Before hospitalization, few or no seizures were reported and IGE had not been diagnosed. IGE was revealed in each patient by a relatively severe event: an absence status (AS), subcontinuous myoclonic seizures or repeated convulsive generalised seizures (CGS). These events were not situation-related but in one patient the relapse of simple convulsive seizures, may have been related to the withdrawal of anti-epileptic drugs (AED) several months previously. EEG records showed generalised spikes or polyspikes and waves organised in a status epilepticus or in interictal rhythmic discharges. In one case they were evident only from a 24 hours recording. Clonazepam injection was used to suppress the AS episode and the subintrant myoclonia. After the AS, interictal generalised epileptic discharges persisted. Two of the four patients had familial history of epilepsy or febrile seizures but in no case was an epileptogenic lesion evident in brain CT scan or MRI. Clinical exams and biologic parameters were normal. All of the patients had worked and were married with children. Appropriate therapies were followed after the diagnosis of IGE. One patient with JME had been treated by Valproate which was discontinued by the general practitioner because of lethargy and replaced by Carbamazepine; seizures were aggravated under both Carbamazepine and then Lamotrigine and until the patient became seizure-free on Levetiracetam. The antiepiletic treatment was also modified in a second patient, while the two others responded well to Valproate. CONCLUSIONS: IGE can exacerbate in the elderly, as different types of seizures including AS, subintrant myoclonia or repeated CGS. Our data suggest persistent IGE are quite frequent in an aged population and may be underestimated due to difficulties in diagnosis. Correctly diagnosed, adjustment of AED may offer substantial clinical improvements in IGE of the elderly. PMID- 19285699 TI - Developing national treatment policy for falciparum malaria in Africa: Malawi experience. AB - The emergence and spread across sub-Saharan Africa of Plasmodium falciparum resistant to the inexpensive antimalarials chloroquine and sulfadoxine pyrimethamine has worsened the health and hampered the socio-economic development of affected countries, a situation that calls for urgent review of malaria treatment policies in these countries. The Roll Back Malaria (RBM) initiative promotes strong partnerships for implementing effective malaria control measures. The development of clear policies to guide such implementation at country level offers a way of assessing the achievement of set milestones in this collaborative venture. In this article we describe the policy development process for the treatment of falciparum malaria in Africa, based on experience in Malawi, where the first-line drug treatment was recently changed from sulfadoxine-pyrimethamine to an artemisinin combination therapy. PMID- 19285701 TI - Polycistronic mRNAs and internal ribosome entry site elements (IRES) are widely used by white spot syndrome virus (WSSV) structural protein genes. AB - The genome of the white spot syndrome virus (WSSV) Taiwan isolate has many structural and non-structural genes that are arranged in clusters. Screening with Northern blots showed that at least four of these clusters produce polycistronic mRNA, and one of these (vp31/vp39b/vp11) was studied in detail. The vp31/vp39b/vp11 cluster produces two transcripts, including a large 3.4-kb polycistronic transcript of all three genes. No monocistronic vp39b mRNA was detected. TNT and in vitro translation assays showed that vp39b translation was independent of vp31 translation, and that ribosomal reinitiation was not a possible mechanism for vp39b. An unusually located IRES (internal ribosome entry site) element was identified in the vp31/vp39b coding region, and this region was able to promote the expression of a downstream firefly luciferase reporter. We show that vp31/vp39b/vp11 is representative of many other WSSV structural/non structural gene clusters, and argue that these are also likely to produce polycistronic mRNAs and that use an IRES mechanism to regulate their translation. PMID- 19285702 TI - Binding of PDZ proteins to HPV E6 proteins does neither correlate with epidemiological risk classification nor with the immortalization of foreskin keratinocytes. AB - There is compelling evidence that high-risk human papillomaviruses (HPV) can cause cervical cancer. Strikingly, HPV16 and 18 account for approximately 70% of all cervical cancers, whereas phylogenetically related types are found at much lower frequencies. Most likely, differences in the activities of the viral E6 and E7 oncoproteins account for the in vivo carcinogenicity. We demonstrate here that E6 proteins from low-risk HPV70 and possibly high-risk HPV82 interact and degrade PDZ proteins hDlg and Magi1 identical to HPV16E6 and HPV18E6. In contrast high risk HPV66E6 did not bind or degrade hDlg or Magi1. We also show that low-risk HPV70 E6/E7 immortalizes normal human keratinocytes. Together with our previous analysis concerning p53 degradation, this shows that neither binding of E6 to p53, to E6AP, to Magi1 and hDlg, the degradation of hDlg and Magi1, nor immortalization of normal human keratinocytes seems to be a reliable predictor for carcinogenic behavior of HPV in the cervix. PMID- 19285703 TI - Biosorption of nickel(II) from aqueous solution by Aspergillus niger: response surface methodology and isotherm study. AB - In the present study, the effects of biosorbent Aspergillus niger dosage, initial solution pH and initial Ni(II) concentration on the uptake of Ni(II) by NaOH pretreated biomass of A. niger from aqueous solution were investigated. Batch experiments were carried out in order to model and optimize the biosorption process. The influence of three parameters on the uptake of Ni(II) was described using a response surface methodology (RSM) as well as Langmuir and Freundlich isotherm models. Optimum Ni(II) uptake of 4.82 mg Ni(II)g(-1) biomass (70.30%) was achieved at pH 6.25, biomass dosage of 2.98 gL(-1) and initial Ni(II) concentration of 30.00 mgL(-1) Ni(II). Langmuir and Freundlich were able to describe the biosorption isotherm fairly well. However, prediction of Ni(II) biosorption using Langmuir and Freundlich isotherms was relatively poor in comparison with RSM approaches. The biosorption mechanism was also investigated by using Fourier transfer infrared (FT-IR) analysis of untreated, NaOH pretreated, and Ni(II) loaded A. niger biomass. PMID- 19285704 TI - Thermally activated peroxydisulfate in the presence of additives: a clean method for the degradation of pollutants. AB - The kinetics and mechanism of the thermal activation of peroxydisulfate, in the temperature range from 60 to 80 degrees C, was investigated in the presence and absence of sodium formate as an additive to turn the oxidizing capacity of the reaction mixture into a reductive one. Trichloroacetic acid, TCA, whose degradation by a reductive mechanism is well reported in the literature, was used as a probe. The chemistry of thermally activated peroxydisulfate is described by a reaction scheme involving free radical generation. The proposed mechanism is evaluated by a computer simulation of the concentration profiles obtained under different experimental conditions. In the presence of formate, SO(4)(-) radicals yield CO(2)(-), which are the main species available for degrading TCA. Under the latter conditions, TCA is more efficiently depleted than in the absence of formate, but otherwise identical conditions of temperature and [S(2)O(8)(2-)]. We therefore conclude that activated peroxydisulfate in the presence of formate as an additive is a convenient method for the mineralization of substrates that are refractory to oxidation, such as perchlorinated hydrocarbons and TCA. This method has the advantage that leaves no toxic residues. PMID- 19285705 TI - Estimating the in situ biodegradation of naphthenic acids in oil sands process waters by HPLC/HRMS. AB - The oil sands industry in Northern Alberta produces large volumes of oil sands process water (OSPW) containing high concentrations of persistent naphthenic acids (NAs; C(n)H(2n+Z)O(2)). Due to the growing volumes of OSPW that need to be reclaimed, it is important to understand the fate of NAs in aquatic systems. A recent laboratory study revealed several potential markers of microbial biodegradation for NAs; thus here we examined for these signatures in field-aged OSPW on the site of Syncrude Canada Ltd. (Fort McMurray, AB). NA concentrations were lower in older OSPW; however parent NA signatures were remarkably similar among all OSPW samples examined, with no discernible enrichment of the highly cyclic fraction as was observed in the laboratory. Comparison of NA signatures in fresh oil sands ore extracts to OSPW in active settling basins, however, suggested that the least cyclic fraction (i.e. Z=0 and Z=-2 homologues) may undergo relatively rapid biodegradation in active settling basins. Further evidence for biodegradation of NAs came from a significantly higher proportion of oxidized NAs (i.e. C(n)H(2n+Z)O(3)+C(n)H(2n+Z)O(4)) in the oldest OSPW from experimental reclamation ponds. Taken together, there is indirect evidence for rapid biodegradation of relatively labile Z=0 and Z=-2 NAs in active settling basins, but the remaining steady-state fraction of NAs in OSPW appear to be very recalcitrant, with half-lives on the order of 12.8-13.6 years. Alternative fate mechanisms to explain the slow disappearance of parent NAs from OSPW are discussed, including adsorption and atmospheric partitioning. PMID- 19285706 TI - Comments on "Interlaboratory comparison of the determination of chlorinated dibenzo-p-dioxins and dibenzofurans according to regulatory methods EN 1948 and EPA 1613b" by M. Wilken et al. [Chemosphere 73 (1) (2008) S2-S6]. PMID- 19285707 TI - Dental microwear texture analysis of two families of subfossil lemurs from Madagascar. AB - This study employs dental microwear texture analysis to reconstruct the diets of two families of subfossil lemurs from Madagascar, the archaeolemurids and megaladapids. This technique is based on three-dimensional surface measurements utilizing a white-light confocal profiler and scale-sensitive fractal analysis. Data were recorded for six texture variables previously used successfully to distinguish between living primates with known dietary differences. Statistical analyses revealed that the archaeolemurids and megaladapids have overlapping microwear texture signatures, suggesting that the two families occasionally depended on resources with similar mechanical properties. Even so, moderate variation in most attributes is evident, and results suggest potential differences in the foods consumed by the two families. The microwear pattern for the megaladapids indicates a preference for tougher foods, such as many leaves, while that of the archaeolemurids is consistent with the consumption of harder foods. The results also indicate some intraspecific differences among taxa within each family. This evidence suggests that the archaeolemurids and megaladapids, like many living primates, likely consumed a variety of food types. PMID- 19285708 TI - Bonobos have a more human-like second-to-fourth finger length ratio (2D:4D) than chimpanzees: a hypothesized indication of lower prenatal androgens. AB - The ratio of the second-to-fourth finger lengths (2D:4D) has been proposed as an indicator of prenatal sex differentiation. However, 2D:4D has not been studied in the closest living human relatives, chimpanzees (Pan troglodytes) and bonobos (Pan paniscus). We report the results from 79 chimpanzees and 39 bonobos of both sexes, including infants, juveniles, and adults. We observed the expected sex difference in 2D:4D, and substantially higher, more human-like, 2D:4D in bonobos than chimpanzees. Previous research indicates that sex differences in 2D:4D result from differences in prenatal sex hormone levels. We hypothesize that the species difference in 2D:4D between bonobos and chimpanzees suggests a possible role for early exposure to sex hormones in the development of behavioral differences between the two species. PMID- 19285709 TI - Factors influencing relationships between personal and ambient concentrations of gaseous and particulate pollutants. AB - Previous exposure studies have shown considerable inter-subject variability in personal-ambient associations. This paper investigates exposure factors that may be responsible for inter-subject variability in these personal-ambient associations. The personal and ambient data used in this paper were collected as part of a personal exposure study conducted in Boston, MA, during 1999-2000. This study was one of a group of personal exposure panel studies funded by the U.S. Environmental Protection Agency's National Exposure Research Laboratory to address areas of exposure assessment warranting further study, particularly associations between personal exposures and ambient concentrations of particulate matter and gaseous co-pollutants. Twenty-four-hour integrated personal, home indoor, home outdoor and ambient sulfate, elemental carbon (EC), PM(2.5), ozone (O(3)), nitrogen dioxide (NO(2)) and sulfur dioxide were measured simultaneously each day. Fifteen homes in the Boston area were measured for 7 days during winter and summer. A previous paper explored the associations between personal-indoor, personal-outdoor, personal-ambient, indoor-outdoor, indoor-ambient and outdoor ambient PM(2.5), sulfate and EC concentrations. For the current paper, factors that may affect personal exposures were investigated, while controlling for ambient concentrations. The data were analyzed using mixed effects regression models. Overall personal-ambient associations were strong for sulfate during winter (p<0.0001) and summer (p<0.0001) and PM(2.5) during summer (p<0.0001). The personal-ambient mixed model slope for PM(2.5) during winter but was not significant at p=0.10. Personal exposures to most pollutants, with the exception of NO(2), increased with ventilation and time spent outdoors. An opposite pattern was found for NO(2) likely due to gas stoves. Personal exposures to PM(2.5) and to traffic-related pollutants, EC and NO(2), were higher for those individuals living close to a major road. Both personal and indoor sulfate and PM(2.5) concentrations were higher for homes using humidifiers. The impact of outdoor sources on personal and indoor concentrations increased with ventilation, whereas an opposite effect was observed for the impact of indoor sources. PMID- 19285710 TI - Bicalutamide demonstrates biologic effectiveness in prostate cancer cell lines and tumor primary cultures irrespective of Her2/neu expression levels. AB - OBJECTIVES: To evaluate the role of Her2/neu as a molecular marker predictive of the treatment response to bicalutamide in prostate cancer (PCa). METHODS: Four PCa cell lines with graded Her2/neu expression levels and 63 primary tumor cultures derived from men with PCa and selected according to Her2/neu tumor levels were used. Primary tumor cultures and PCa cell lines were treated with bicalutamide (0.1-10 microM) in the presence of dehydrotestosterone (10(-12) M) for 4 days. The presence of a significant correlation between Her2/nue expression and drug efficacy was used to define the role of Her2/neu as molecular predictor of bicalutamide effectiveness. As an indicator of drug effectiveness we used the concentration that inhibits 50% values determined after bicalutamide treatment. RESULTS: After bicalutamide treatment, no significant differences in the concentration that inhibits 50% were found among the different tumor cell lines (P = .081). In this experimental model, the correlation analysis suggested that the effectiveness of this drug was not influenced by Her2/neu levels (r = 0.053, P = .823). In primary cultures with high Her2/neu levels (43 tumor cultures), the mean value of the concentration that inhibits 50% for bicalutamide was 0.43 +/- 0.13 microM, and in cultures with low Her2/neu levels (20 tumor cultures), the same parameter was 0.5 +/- 0.16 microM (P = .14). The correlation analysis suggested that the effectiveness of this drug was not influenced by Her2/neu levels (r = 0.33, P = .101). CONCLUSIONS: Our biologic data seem to indicate that the antitumor effect of bicalutamide is independent of Her2/neu levels in preclinical models of PCa. Bicalutamide could be configured as a pharmacologic option to treat patients with high or low levels of Her2/neu. PMID- 19285711 TI - Dendritic cells with transduced survivin gene induce specific cytotoxic T lymphocytes in human urologic cancer cell lines. AB - OBJECTIVES: To investigate whether survivin-specific cytotoxic T lymphocytes (CTLs) could be induced by dendritic cells (DCs) transduced with survivin gene by adenoviral vector, and whether these CTLs would display cytotoxic activities against human urologic cancer cell lines. Survivin, a member of the inhibitor of apoptosis protein family, is expressed in most malignancies, but not in normal tissue. METHODS: Adenoviral vector encoding the human survivin gene was generated. Human DCs from healthy donors were transduced with human survivin gene by infection with adenoviral vector encoding the human survivin gene using the centrifugal method. Survivin-specific CTLs were induced from autologous peripheral blood mononuclear cells by DCs transduced with the survivin gene. The ability of CTLs to lyse cancer cell lines was assessed using the (51)Cr-release assay. RESULTS: DCs transduced with survivin gene could induce survivin-specific CTLs against various urologic malignancies such as bladder, kidney, and prostate cancer cells. This cytotoxic activity could be blocked by anti-CD8 and anti-major histocompatibility complex class I antibodies. We also found that this cytotoxic activity was specific for the survivin protein and human leukocyte antigen haplotype. CONCLUSIONS: DCs transduced with the survivin gene induced potent survivin-specific CTL responses in vitro. This suggests that cancer immunotherapy targets for survivin might offer a novel approach to treating various urologic cancers. PMID- 19285712 TI - Presacral extrarenal Wilms tumor in a child. AB - OBJECTIVES: Wilms tumor (WT) is one of the most common solid tumors in children. It also is the most frequent tumor found in the kidneys. In 5% of cases, it affects both kidneys simultaneously. Extrarenal WTs are extremely rare. The diagnosis of extrarenal WT is almost always made after surgical intervention. The tumor can be located in the retroperitoneum, uterus, cervix, testes, skin, and even the thorax. METHODS: We report on a 3-year-old girl with an extrarenal WT located in the presacral region. RESULTS: The exact mechanism whereby a WT occurs in extrarenal tissues is unknown. The presence of ectopic metanephric blastema cells or the WT gene causing transformation of extrarenal primitive mesonephric or pronephric remnants into WT have both been considered in the embryogenesis. Although imaging examinations are helpful in the definition of retroperitoneal tumors, no characteristic findings are available to diagnose an extrarenal WT before surgery. CONCLUSIONS: As a rule, the diagnosis of extrarenal WT is made after surgery. Surgical excision is the treatment of choice and the same general therapeutic rules should be followed as when the kidney has been affected. The use of Stage III guidelines for chemotherapy and radiotherapy are appropriate for these patients. PMID- 19285713 TI - Risk factors affecting cure after mid-urethral tape procedure for female urodynamic stress incontinence: comparison of retropubic and transobturator routes. AB - OBJECTIVES: To identify the risk factors affecting cure after the tension-free vaginal tape procedure (TVT) or transobturator tape procedure (TOT). METHODS: A total of 121 women with urodynamically proved stress incontinence underwent TVT (n = 61) or TOT (n = 60). We analyzed all data with univariate and multivariate logistic regression analyses. RESULTS: Univariate analysis revealed that a larger postvoid residual urine volume, greater pad weight test, and lower maximal flow rate were associated with a greater risk of failure to be cured after TVT. A lower maximal urethral closure pressure (MUCP) was associated with a greater risk of failure after the TOT procedure. With additional analysis, we found that a MUCP of 1000 laparoscopic procedures, we have used more or less complex catheterization techniques. After developing the use of the nephroscopy needle trocar for laparoscopic urinary diversion and finding that such use simplified the procedure, this technique was extended to all other laparoscopic procedures in which catheterization is required. RESULTS: We analyzed the use of the nephroscopy needle trocar in 15 ureteropyeloplasty, 21 ileal conduit, and 4 laparoscopic ureteral reimplantation procedures. The mean operating time required for anterograde catheterization in ureteropelvic junction stenosis was 3 minutes, 35 seconds, and the mean operating time for retrograde catheterization was 2 minutes, 20 seconds. Anterograde catheterization was impossible in 1 case. No cases of a false ureteral tract occurred. CONCLUSIONS: The results of our analysis have shown that the use of the nephroscopy needle trocar for percutaneous catheterization in laparoscopic surgery markedly simplifies the procedure, with the resultant savings in operating time. PMID- 19285717 TI - Accuracy of in vivo and ex vivo ultrasonographic evaluation of ovarian follicles and corpora lutea in sows. AB - Current study determined, in sows, the accuracy of ultrasonography for in vivo (n=8) and ex vivo (n=7) evaluation of corpora lutea (CLs) and follicles > or =1.5mm in size, by comparison with macroscopic findings in sliced ovaries. The accuracy for ex vivo detection of follicles increased with follicle size (P<0.05), being low for 1.5-1.9 mm follicles (65.9%) and higher for > or =6mm follicles (93.3%); differences between ultrasonographic and macroscopic observations were significant only for follicles smaller than 3.9 mm (P<0.05), due to underestimation. Ex vivo observation succeeded to detect presence or absence of CLs in all the ovaries; the efficiency for determining the exact number of CLs being 94.4%. The accuracy for in vivo detection of follicles also increased with follicle size (P<0.05), dropping to values lower than 40% for 1.5 1.9 mm follicles; therefore, there were significant differences between ultrasonographic and macroscopic observations (P<0.05). On the other hand, accuracy remained around 92% for > or =6mm follicles. Ultrasonography was useful again for detecting presence of CLs in all the ovaries; the efficiency for determining CLs number reached 86.7%, due to underestimation in ovaries with higher number of CLs (P<0.05). Overall, there were no significant differences when comparing the accuracy of ex vivo and in vivo scannings for determination neither of the number of follicles in each size-category larger than 1.9 mm nor of the presence of ovulations or of the CLs number in each ovary. In conclusion, the use of ultrasonography allows an accurate detection of the presence and number of CLs and follicles > or =2mm of diameter in sows, without significant differences between in vivo and ex vivo observations. PMID- 19285718 TI - Paclitaxel delivery from PLGA foams for controlled release in post-surgical chemotherapy against glioblastoma multiforme. AB - Paclitaxel loaded biodegradable poly-(DL-lactic-co-glycolic) acid (PLGA) foams with microporous matrix were fabricated by a novel pressure quenching approach to provide a sustained paclitaxel release. The foams with micropores provided increased surface area to volume ratio and were also implantable for post surgical chemotherapy applications. The two formulations 5% (w/w) paclitaxel loaded PLGA 85:15 foam (F1) and 10% (w/w) paclitaxel loaded PLGA 50:50 foam (F2), were evaluated in vitro and in vivo. Both the foams were found to provide a paclitaxel release beyond a month in vitro with a near zero-order kinetics and with minimum burst release. Furthermore, apoptosis of C6 glioma cells in vitro demonstrated the benefits of sustained paclitaxel release by the foams in comparison to acute Taxol exposure. Both the foams exhibited continuous paclitaxel release in an in vivo (subcutaneous) environment up to a month which correlated well with the in vitro release profiles. Bio-distribution results in the rat brain showed paclitaxel penetration at therapeutic levels up to 3mm into the tissue from the site of foam implantation. Hence these foams could be employed as potential implants for post-surgical chemotherapy against malignant glioma. PMID- 19285719 TI - Fluorescent microparticles for sensing cell microenvironment oxygen levels within 3D scaffolds. AB - We present the development and characterization of fluorescent oxygen-sensing microparticles designed for measuring oxygen concentration in microenvironments existing within standard cell culture and transparent three-dimensional (3D) cell scaffolds. The microparticle synthesis employs poly(dimethylsiloxane) to encapsulate silica gel particles bound with an oxygen-sensitive luminophore as well as a reference or normalization fluorophore that is insensitive to oxygen. We developed a rapid, automated and non-invasive sensor analysis method based on fluorescence microscopy to measure oxygen concentration in a hydrogel scaffold. We demonstrate that the microparticles are non-cytotoxic and that their response is comparable to that of a traditional dissolved oxygen meter. Microparticle size (5-40 microm) was selected for microscale-mapping of oxygen concentration to allow measurements local to individual cells. Two methods of calibration were evaluated and revealed that the sensor system enables characterization of a range of hypoxic to hyperoxic conditions relevant to cell and tissue biology (i.e., pO(2) 10-160 mmHg). The calibration analysis also revealed that the microparticles have a high fraction of quenched luminophore (0.90+/-0.02), indicating that the reported approach provides significant advantages for sensor performance. This study thus reports a versatile oxygen-sensing technology that enables future correlations of local oxygen concentration with individual cell response in cultured engineered tissues. PMID- 19285720 TI - Engineering thermoresponsive polymeric nanoshells. AB - This study describes a family of hollow nanoscale constructs able to display a large and reversible change in size, within a narrow temperature interval. These thermoresponsive nanostructures are generated by crosslinking functionalized amphiphilic molecules, such as poly(ethylene oxide)-poly(propylene oxide) poly(ethylene oxide) (PEO-PPO-PEO) triblocks, while constrained to their particular micellar configuration. This contribution focuses on the generation of these supramolecular architectures, by intra-micellarly crosslinking PEO-PPO-PEO dimethacrylate amphiphiles, and investigates their temperature-dependent dimensional behavior. Spherical nanoshells displaying a 200 nm diameter at 15 degrees C, shrink sharply around 28 degrees C, generating a compact structure of approximately 40 nm at body temperature. Thermoresponsive nanotubes were created by generating them from rod-like micelles at higher temperatures. By varying the composition of the triblock, the transition temperature can be fine tuned, from around 25 degrees C to slightly above body temperature. Also, the nanoshells were rendered biodegradable by incorporating aliphatic oligoesters into their structure. Numerous applications for these nanostructures are foreseen in various biomedical areas, such as in drug and gene delivery, in the tissue engineering and in the biosensors field. PMID- 19285721 TI - A key role for dense granule secretion in potentiation of the Ca2+ signal arising from store-operated calcium entry in human platelets. AB - Recent work has demonstrated a role for Na(+)/Ca(2+) exchange in potentiation of the Ca(2+) entry elicited through the human platelet store-operated channel by controlling a Mn(2+)-impermeable Ca(2+) entry pathway. Here we demonstrate that this involves control over the secretion of dense granules by a Na(+)/Ca(2+) exchanger (NCX) and so autocrine signalling between platelets. NCX inhibition reduced dense granule secretion. The reduction in SOCE elicited by NCX inhibition could be reversed by the addition of uninhibited donor cells, their releasate alone, or exogenous ADP and 5-HT. The use of specific receptor antagonists indicated that ATP, ADP and 5-HT all played a role in NCX-dependent autocrine signalling between platelets following thapsigargin stimulation, by activating Mn(2+)-impermeable Ca(2+) entry pathways. These data provide further insight into the mechanisms underlying the known interrelationship between platelet Ca(2+) signalling and dense granule secretion, and suggest an important role for the NCX in potentiation of platelet activation via dense granule secretion and so autocrine signalling. Our results caution the interpretation of platelet Ca(2+) signalling studies involving pharmacological or other manipulations that do not assess possible effects on NCX activity and dense granule secretion. PMID- 19285722 TI - The UPR as a survival factor of cancer cells: More than folding proteins? PMID- 19285723 TI - Primary Epstein-Barr virus-negative nasal-type natural killer/T cell lymphoma of the testis. PMID- 19285724 TI - Adenovirus-associated hemorrhagic cystitis in a patient with plasma cell myeloma treated with bortezomib. PMID- 19285725 TI - Repeated reports for child maltreatment among intimate partner violence victims: findings from the National Survey of Child and Adolescent Well-Being. AB - OBJECTIVES: To determine the prevalence of intimate partner violence (defined as any physical violence during the last 12 months or previously) among mothers who maltreat their children, and to examine whether mothers' experiences of intimate partner violence (IPV) are associated with repeated reports (rereports) of children to Child Protective Services (CPS) during the following 18 months. METHODS: Data for the analyses were from the National Survey of Child and Adolescent Well-Being (NSCAW), a national probability study of children investigated for child maltreatment. The sample of 5,501 children (ages 0-14) was randomly selected from the families who entered the US child welfare system between October 1999 and December 2000. The analysis sample was restricted to 1,236 families in which caregivers were: (1) the alleged perpetrators of the child maltreatment at baseline (independently of substantiation status) and (2) the biological mothers (n=1,212 or 98.6%), adoptive mothers (n=17 or 1%), or stepmothers (n=7 or 0.3%) of children not placed in out-of-home care. RESULTS: Children of mothers physically abused by an intimate partner during the last 12 months or previously at the intake interview (44%) were twice as likely as children of mothers who had not experienced such violence to be rereported to CPS (29% vs. 14%, Odds Ratio=2.0, 95% Confidence Interval=1.1-3.4). Rereports occurred almost twice as quickly for children of mothers who experienced IPV compared to children of mothers who had not experienced IPV (Hazard Ratio=1.9, 95% Confidence Interval=1.1-3.0). CONCLUSIONS: The higher risk and speedier rereports of child maltreatment associated with intimate partner violence highlights the need for universal assessment and provision of services for IPV among families that are investigated by CPS. PMID- 19285726 TI - The Allium cepa bioassay to evaluate landfarming soil, before and after the addition of rice hulls to accelerate organic pollutants biodegradation. AB - Landfarming is a soil bioremediation technology practiced by oil refineries in order to reduce or eliminate hydrocarbons from petroleum sludge. The goal of the current study was to use Allium cepa bioassay to assess landfarming and landfarming with rice hulls amendment before and after hydrocarbons biodegradation assay in the laboratory. Three cytogenetic endpoints were used: mitotic and chromosome abnormalities (MCA), micronucleus (MN) and nuclear buds (NB). Landfarming presented 13.5 g/kg of total petroleum hydrocarbons (TPHs) and caused strong clastogenic and mutagenic effects (p<0.05) in A. cepa. After 108 days of biodegradation, the landfarming reached the rate of 26.30 mmol of CO(2) released, the concentration of TPHs decreased by 27% and there was significant reduction in MCA, MN and NB. Landfarming treated with rice hulls had the highest release of CO(2), 110.9 mmol, associated with a remarkable reduction in TPHs concentration, 59%, and had the highest decrease in MCA, MN and NB (p>0.05). Our findings showed that the use of rice hulls accelerated the biodegradation efficacy of landfarming and reduced their clastogenicity, indicating that supplementary treatments are important to improve the efficiency of bioremediation processes. PMID- 19285727 TI - An assessment of the tolerance of Ligustrum ovalifolium Hassk. to traffic generated Pb using physiological and biochemical markers. AB - This study examined the leaf Pb content, chlorophyll fluorescence parameters (Fo, Fm, Fv, Fv/Fm, Fm/Fo and t(1/2)), photopigments (Chl a, Chl b, Chl a+b, the Chl a/b ratio, and total carotenoids), as well as total phenolics (free and bound) in privet (Ligustrum ovalifolium Hassk.) in 'Stara Zvezdara' Park, located along the avenue with heavy traffic flow (polluted site), and the Arboretum of Belgrade's Faculty of Forestry (control site). Site-dependent variations were observed for all the parameters examined. It was found that Pb accumulated in leaf tissues (3.5-4.2microg/g) originates from traffic. Results obtained showed that privet is tolerant to the accumulation of Pb generated by traffic, because it maintains optimal photosynthesis and is characterized by active protection mechanisms due to increase in leaf phenolics, which enable it to survive in habitats exposed to chronic Pb pollution stress. Therefore, L. ovalifolium has great potential for urban landscaping. PMID- 19285728 TI - Quality improvements in the early care and education workforce: outcomes and impact of the T.E.A.C.H. Early Childhood Project. AB - To date there has not been a systematic, longitudinal research to assess the efficacy of public investments in the professional development of early childhood educators that are funded through the T.E.A.C.H. Early Childhood Project. This study of the T.E.A.C.H. Early Childhood Project Pennsylvania provides a longitudinal analysis of the data for the first three cohorts of applicants and scholars that participated in Pennsylvania's T.E.A.C.H. program. Over a 5-year period, we followed the scholars that participated in this program to examine the direct outcomes, as well as the indirect impact, of participating in the T.E.A.C.H. scholarship program. We found a number of positive outcomes for the scholars with respect to: educational achievement, increased compensation, reduced turnover, and improved knowledge, skill, and practice. The downside of T.E.A.C.H., as implemented in Pennsylvania, is keeping caregivers in the program. Only 15% of the scholars were still in the program at the end of the 5th year. The dropout/withdrawal rate after the first contract was 43.5% representing a sizable investment in scholars that never completed the initial contract. Based on these findings, the implications policy and practice are discussed. PMID- 19285729 TI - Stage specific over-expression of the dominant negative Ikaros 6 reveals distinct role of Ikaros throughout human B-cell differentiation. AB - Ikaros is a transcription factor that acts both as an activator and as an inhibitor of gene expression in several hematopoietic lineages. Ikaros functions in hematopoiesis have mostly been studied in mice, and are notably crucial for lymphopoiesis. Deregulation of Ikaros expression was evidenced in several leukemia subtypes, including pre-B-ALL. Here, we studied the role of Ikaros in human B lymphoid differentiation through xeno-transplantation of genetically modified cord blood (CB) human hematopoietic progenitor cells (HPC) in NOD/SCID mice. We used lentiviral vectors to force expression of Ikaros 6 (Ik6), a known dominant negative (DN) protein that interferes with normal Ikaros and structurally related proteins in HPC and their progeny. Two types of vectors were used: a vector containing the EF1alpha promoter which produces strong gene expression in all hematopoietic lineages, and a recently validated B-specific vector containing an enhanced CD19 derived promoter that strongly favors expression in the B-cell lineage. Ik6 transduction of CB CD34(+) cells with these vectors produced distinct consequences in the B-cell differentiation profiles of xenografted human cells. While the ubiquitous vector favored a specific block at the early pro-B/pre-B stage of differentiation, with an increase in Lambda Like transcript expression in the bone marrow (BM), B-specific Ik6 expression provoked a global decrease in the CD19(+) cell population in both BM and spleen, associated with a decrease in IgM+ immature B-cells in the spleen. We conclude that Ikaros proteins are active throughout human B-cell differentiation, before and after CD19 appearance. PMID- 19285730 TI - The cost-effectiveness of routine office-based identification and subsequent medical treatment of primary open-angle glaucoma in the United States. AB - OBJECTIVE: To estimate the incremental cost-effectiveness of routine glaucoma assessment and treatment under current eye care visit and treatment patterns and different levels of treatment effectiveness (from randomized trials). DESIGN: We compared the costs and benefits of routine glaucoma assessment and treatment compared with no treatment using conservative and optimistic assumptions regarding treatment efficacy and including and excluding prediagnostic assessment costs. PARTICIPANTS AND CONTROLS: Computer simulation of 20 million people followed from age 50 years to death or age 100 years. METHODS: With the use of a computer model, we simulated glaucoma incidence, natural progression, diagnosis, and treatment. We defined glaucoma incidence conservatively as a mean deviation of -4 decibels (dB) on visual field testing in either eye for all diagnoses to be both clinically meaningful and unambiguous. We simulated the annual probability of subsequent progression and the quantity of visual field lost when progression occurred. MAIN OUTCOME MEASURES: Visual field loss, ophthalmologic and nursing home costs, quality-adjusted life years (QALYs), cost per QALY gained, and cost per year of sight gained. Costs and QALYs were discounted to 2005 values using a 3% rate. RESULTS: Compared with no treatment and when including diagnostic assessment costs, the incremental cost-effectiveness of routine assessment and treatment was $46,000 per QALY gained, assuming conservative treatment efficacy, and $28,000 per QALY gained, assuming optimistic treatment efficacy. Compared with no treatment and when excluding diagnostic assessment costs, the incremental cost-effectiveness of routine assessment and treatment was $20,000 per QALY gained, assuming conservative treatment efficacy, and $11,000 per QALY gained, assuming optimistic treatment efficacy. The cost-effectiveness was most sensitive to the treatment costs and the value of QALY losses assigned to visual field losses. CONCLUSIONS: Glaucoma treatment was highly cost-effective when the costs of diagnostic assessments were excluded or when we assumed optimistic treatment efficacy. The cost was reasonable and in line with other health interventions even when diagnostic assessment costs were included and assuming conservative efficacy. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article. PMID- 19285731 TI - Endobronchial ultrasound-guided biopsy in the evaluation of intrathoracic lymphadenopathy in suspected tuberculosis: a minimally invasive technique with a high diagnostic yield. PMID- 19285732 TI - The burden of awareness of psychometric risk for schizophrenia. AB - Participants in studies of psychometric risk for schizophrenia are rarely informed of their risk status. Nondisclosure may be justifiable if the harmful effects of disclosure outweigh its benefits. We examined whether disclosure may adversely affect well-being and, if so, factors that predict the degree of adverse effect. Undergraduates (n=114) rated the anticipated impact--on felt distress, coping, optimism, helplessness, future lifestyle choices, and survival- of discovering they were at risk for schizophrenia and six other diseases. They also completed measures of potential predictors of this impact, including knowledge about schizophrenia, vicarious experience of schizophrenia, their potential to suffer stigmatization because of schizophrenia, and schizotypy. Participants judged schizophrenia risk more negatively than risk for heart disease, arthritis, depression, and diabetes, and less negatively than risk for cancer and Alzheimer's disease. Higher disorder-nonspecific impact, greater stigma, and lower psychometric risk for schizophrenia together provided the best linear prediction of schizophrenia-specific impact. Awareness of schizophrenia risk creates a significant adverse impact, the level of which may be greatest among those with lowest risk. PMID- 19285733 TI - The influence of age and Rhodococcus equi infection on CD1 expression by equine antigen presenting cells. AB - There is a distinct age-associated susceptibility of horses to Rhodococcus equi infection. Initial infection is thought to occur in the neonatal and perinatal period, and only foals less than 6 months of age are typically affected. R. equi is closely related and structurally similar to Mycobacterium tuberculosis, and causes similar pathologic lesions. Protective immune responses to M. tuberculosis involve classical major histocompatibility complex (MHC)-restricted T cells that recognize peptide antigen, as well as MHC-independent T cells that recognize mycobacterial lipid antigen presented by CD1 molecules. Given the structural similarity between these two pathogens and our previous observations regarding R. equi-specific, MHC-unrestricted cytotoxic T lymphocytes (CTL), we developed 3 related hypotheses: (1) CD1 molecules are expressed on equine antigen presenting cells (APC), (2) CD1 expression on APC is less in foals compared to adults and (3) infection with live virulent R. equi induces up-regulation of CD1 on both adult and perinatal APC. CD1 expression was examined by flow cytometric analysis using a panel of monoclonal CD1 antibodies with different species and isoform specificities. RESULTS: Three CD1 antibodies specific for CD1b showed consistent cross reactivity with both foal and adult monocyte-derived macrophages (MDM). CD1b and MHC class II expression were significantly higher on adult MDM compared with foals. R. equi infected MDM showed significantly lower expression of CD1b, suggesting that infection with this bacterium induces down-regulation of CD1b on the cell surface. Histograms from dual antibody staining of peripheral blood mononuclear cells also revealed that 45-71% of the monocyte population stained positive for CD1b, and that the majority of these also co-expressed MHC II molecules, indicating that they were APC. The anti-CD1 antibodies showed no binding or minimal binding to bronchoalveolar lavage (BAL)-derived macrophages. CONCLUSION: The CD1b isoform is evolutionarily conserved, and is present on equine MDM, as well as on circulating blood monocytes. The unique susceptibility of foals to R. equi infection may be due in part to lower expression of CD1 and MHC class II, as observed in this study. The data also suggests that infection with R. equi induces down-regulation of CD1b on equine MDM. This may represent a novel mechanism by R. equi to avoid detection and killing of infected cells by the immune system, similar to that observed when human APC are infected with M. tuberculosis. PMID- 19285734 TI - Rate of eradication of group A beta-hemolytic streptococci in children with pharyngo-tonsillitis by amoxicillin and cefdinir. AB - BACKGROUND: Cephalosporins were found to be more effective than penicillins in the eradication of group A beta-hemolytic streptococcal (GABHS) from tonsillar tissues. This study investigated the effect of amoxicillin and cefdinir therapies on the rate of eradication of GABHS from the tonsils of children with acute pharyngo-tonsillitis (PT). PATIENTS AND METHODS: Of 50 children suffering from PT 25 were treated with amoxicillin (40 mg/(kg d) or 250 mg every 8 h) and 25 with cefdinir (14 mg/(kg d) or 600 mg once a day) for 10 days. Pharyngo-tonsillar cultures were obtained from all children before treatment and on the 1st, 2nd, 3rd, 4th, 7th, and 12th days. RESULTS: GABHS was eradicated more rapidly from children treated with cefdinir as compared to those receiving amoxicillin. A smaller number of patients with GABHS were found in those treated with cefdinir as compared to amoxicillin throughout the treatment period. Eradication of GABHS from 68% (8 of 25 patients) was noted in those treated with cefdinir after 2 days and those treated with amoxicillin after 4 days. The differences between the number of patients who had a bacteriological cure between those receiving cefdinir to those getting amoxicillin was statistical significant at day 4 (32% vs. 8%). At the end of therapy GABHS was recovered from 5 (20%) and 2 (8%) of the patients. The group that received cefdinir, had a more rapid reduction in fever on the first after initiation of therapy as compared to those receiving amoxicillin. The fever reduction in those receiving cefdinir occurred a day earlier than in those getting amoxicillin. CONCLUSIONS: Fever was reduced and GABHS was eradicated more rapidly from children treated with cefdinir as compared to amoxicillin. PMID- 19285735 TI - Subcellular neuropharmacology: the importance of intracellular targeting. AB - Few cell types are more adapted for cell-cell signaling than neurons. Their responsiveness lies in the formation of highly specialized compartments composed of unique repertoires of selectively distributed protein complexes generated, in part, by the local translation of mRNAs and regulated by their RNA-binding proteins. Utilizing the selective distribution of these neuronal proteins and the underlying mechanisms that generate the differential patterns of expression as central facets of drug design promises to enhance the therapeutic ratio of a drug. It is in this context that we discuss the unique arrangement of mRNAs, RNA binding proteins and the protein macromolecular complexes at the dendrite, which is the postsynaptic site of synaptic transmission. Recent advances in identifying the function of dendritic components of the mechanisms of protein and RNA transport, non-nuclear RNA splicing and localized translation underscore their importance as targets of neuropharmacology. PMID- 19285736 TI - Hot flash: TRPV channels in the brain. AB - TRPV1 (transient receptor potential, vanilloid) channels belong to a family of ligand-gated ion channels gated not only by the binding of certain lipophilic molecules but also by extracellular protons and physical stimuli such as heat or osmotic pressure changes. These nonselective cation channels are permeable to Na(+) and K(+) and are also very Ca(2+) permeable; in fact, TRPV1 is as Ca(2+) permeable as the NMDA receptor channel and can, thus, act as a trigger for Ca(2+) mediated cell signaling. Although these channels are highly expressed in primary sensory afferents, accumulating evidence indicates that TRPV family channels are also present in the brain. Here, we review evidence that TRPV channels in the central nervous system might contribute to many basic neuronal functions including resting membrane potential, neurotransmitter release and synaptic plasticity. PMID- 19285737 TI - Autopsy reports in extreme longevity. AB - This study aimed at establishing the prevalence, the type, the severity of various diseases, as well as the main causes of death in the elderly and ultralongevous subjects. The autopsy findings of 140 centenarians (21 males and 119 females) of the age range of 100-109 years were compared to those of 96 elderly subjects (14 males and 82 females) of the age range of 75-95 years. In all cases the clinical diagnosis, the clinical record, the macro- and microscopic findings and the autopsy protocols were evaluated. A lower prevalence (16.3% vs. 39.0%), as well as a slower and less aggressive evolution of neoplastic pathologies (frequency of metastases: 26.0% vs. 55.0%) in the centenarians, as compared to the general aging population, have been found. The chronic degenerative pathologies, especially the cerebro-degenerative ones were observed more frequently and were of major gravity in the centenarians, compared to the elderly population. The cerebrovascular damage and the consequent cognitive deficit do not influence the survival of the longevous subjects. Intercurrent events or external accidents may interrupt the weak equilibrium of these "frail" subjects. PMID- 19285738 TI - Association of P wave duration and dispersion with the risk for atrial fibrillation: practical considerations in the setting of coronary artery disease. AB - P wave dispersion (PWD) is defined as the difference between maximum P wave duration (Pmax) and minimum P wave duration recorded from multiple surface electrocardiogram (ECG) leads. An increase in PWD indicates heterogeneous intraatrial and interatrial conduction and discontinuous anisotropic propagation of sinus impulses, providing a substrate that favors reentry mechanisms. Prolonged Pmax and increased PWD have been suggested to represent independent predictors for atrial fibrillation (AF). The distinctive atrial electrophysiological peculiarity featured by slow interrupted propagation of atrial impulses could be reflected by an increase in PWD predisposing to AF. Coronary artery disease (CAD) is also a well-known risk factor for AF. There are some potential mediators between CAD and AF. The noninvasive nature of P wave duration and PWD with their comparative simplicity makes these parameters attractive options; however, standards involving precise assessment are still lacking. Further exploration of the exact determinants for P wave duration and PWD would help establish the practical usefulness of these surface ECG markers in estimating the risk for AF occurrence and recurrence. PMID- 19285739 TI - Reduced regional systolic function evolved compacted segments in noncompaction. PMID- 19285741 TI - The influence of Thai culture on diabetes perceptions and management. AB - OBJECTIVE: To explore the way Thai patients perceive and manage their diabetes. RESEARCH DESIGN AND METHODS: Using a focused ethnographic approach, face-to-face interviews were conducted at two public hospitals in Bangkok. All interviews (n=27) were audio-taped and transcribed verbatim. Analysis of the interview transcripts was completed thematically. RESULTS: The findings showed that Thai patients manage their diabetes according to their beliefs about diabetes. These beliefs are constructed using both modern and traditional knowledge. For example, some patients explained the cause of their illness as being due to biomedical factors such as genetics, and also cultural factors such as karma from either previous or current lifetimes. The analysis also revealed that some aspects of Thai life facilitate diabetes self-management while other aspects hamper good control of the illness. For example, Buddhist values of moderation contribute positively to dietary change, while, on the other hand, the importance of rice in the Thai diet can impede successful self-management strategies. CONCLUSION: The results of this research indicate that Thai culture influences diabetes perceptions and management. Culturally appropriate treatment guidelines should be established for diabetes management that give special consideration to the significance and meaning of food and to Buddhist beliefs. PMID- 19285743 TI - Hepatitis D: thirty years after. AB - The key to the discovery of the Hepatitis D Virus (HDV) was the description in Turin, Italy in the mid-1970s of the delta antigen and antibody in carriers of the hepatitis B surface antigen. The new antigen was first thought to be a marker of the Hepatitis B Virus (HBV) and in view of its intricate true nature, it would have possibly died away as another odd antigenic subtype of HBV, like many that were described in the 1970s. Fortunately, instead, a collaboration started in 1978 between the Turin group, and the National Institute of Health and Georgetown University in the US. With American facilities and expertise this collaboration led just a year later, in 1979, to the unfolding of an unexpected and amazing chapter in virology. Experiments in chimpanzees demonstrated that the delta antigen was not a component of the HBV but of a separate defective virus requiring HBV for its infection; it was named the hepatitis D virus to conform to the nomenclature of hepatitis viruses and classified within the genus Deltavirus. The animal experiments were also seminal in proposing to future clinical interpretation, the paradigm of a pathogenic infection (hepatitis D), that could develop only in HBV-infected patients, was mainly transmitted by superinfection of HDV on chronic HBV carriers and had the ability to strongly inhibit the helper HBV. The discovery of the HDV has driven three directions of further research: (1) The understanding of the replicative and infectious mechanisms of the HDV. (2) The assessment of its epidemiological and medical impact. (3) The search for a therapy for chronic hepatitis D (CHD). This review summarizes the progress achieved in each field of research in the thirty years that have passed since the discovery of HDV. PMID- 19285745 TI - Evolution of the achaete-scute complex in insects: convergent duplication of proneural genes. AB - Proneural genes encode transcriptional activators of the basic Helix-loop-helix class that are involved in neuronal specification and differentiation. We have used the recent availability of genome sequences of multiple distant insect species to study the evolution of a family of proneural genes, the achaete-scute genes, and to examine their genomic organization and evolution. We document independent evolution of multiple copies of achaete-scute homologues and argue that this might have contributed to morphological diversity in Diptera and Lepidoptera. PMID- 19285746 TI - The complex relationship of gene duplication and essentiality. AB - In yeast and worm, duplicate genes overlap in function so that deleting one of a pair from the genome is less likely to be lethal than deleting a singleton gene. By contrast, previous analyses showed that mouse duplicate genes were as essential as singletons. We show that the relationship between gene duplication and essentiality is complex in multicellular organisms, with developmental genes and genes that were duplicated by whole genome duplication being more essential than other duplicated genes. PMID- 19285747 TI - An event-driven distributed processing architecture for image-guided cardiac ablation therapy. AB - Medical imaging data is becoming increasing valuable in interventional medicine, not only for preoperative planning, but also for real-time guidance during clinical procedures. Three key components necessary for image-guided intervention are real-time tracking of the surgical instrument, aligning the real-world patient space with image-space, and creating a meaningful display that integrates the tracked instrument and patient data. Issues to consider when developing image guided intervention systems include the communication scheme, the ability to distribute CPU intensive tasks, and flexibility to allow for new technologies. In this work, we have designed a communication architecture for use in image-guided catheter ablation therapy. Communication between the system components is through a database which contains an event queue and auxiliary data tables. The communication scheme is unique in that each system component is responsible for querying and responding to relevant events from the centralized database queue. An advantage of the architecture is the flexibility to add new system components without affecting existing software code. In addition, the architecture is intrinsically distributed, in that components can run on different CPU boxes, and even different operating systems. We refer to this Framework for Image-Guided Navigation using a Distributed Event-Driven Database in Real-Time as the FINDER architecture. This architecture has been implemented for the specific application of image-guided cardiac ablation therapy. We describe our prototype image guidance system and demonstrate its functionality by emulating a cardiac ablation procedure with a patient-specific phantom. The proposed architecture, designed to be modular, flexible, and intuitive, is a key step towards our goal of developing a complete system for visualization and targeting in image-guided cardiac ablation procedures. PMID- 19285748 TI - Interactive image analysis programs for quantifying injury-induced axonal beading and microtubule disruption. AB - Focal axonal beading and focal disruption of microtubule structure are characteristic to traumatic axonal injury. We have recently reproduced these morphological and structural changes in our in vitro model system [D. Kilinc, G. Gallo, K.A. Barbee, Mechanically induced membrane poration causes axonal beading and localized cytoskeletal damage, Exp. Neurol. 212 (2008) 422-430]. In order to measure bead formation objectively, an observer-independent quantification of beading was necessary. In addition, a quantitative measure for the extent of co localization of axonal beads and microtubule disruptions was required to establish a causal relationship between focal cytoskeletal damage and bead formation. In this paper we describe Matlab-based, interactive image analysis programs for axonal beading quantification and co-localization analysis. Injury induced increases in the axonal beading could be successfully detected using the bead analysis program. PMID- 19285749 TI - Osteopontin is involved in the development of acquired chemo-resistance of cisplatin in small cell lung cancer. AB - Osteopontin (OPN) is a multi-functional cytokine involved in cell survival, migration and adhesion which is associated with tumorigenesis, progression and metastasis. However, the role of OPN in chemo-sensitivity of human lung cancer has not yet been elucidated. The purpose of this study is to investigate the role of OPN in chemo-sensitivity of lung cancer cells. We developed a stable OPN transfectant (SBC-3/OPN) and a control transfectant (SBC-3/NEO) from human small cell lung cancer cell line, SBC-3. SBC-3/OPN cells were more resistant to cisplatin than SBC-3/NEO cells. Multi-drug resistance-associated protein (MRP) does not appear to be involved in the development of acquired chemo-resistance, since MRP inhibitor did not alter chemo-sensitivity. After exposure to cisplatin, the apoptotic SBC-3/OPN cells were reduced in number compared to SBC-3/NEO cells. Treatment with cisplatin revealed that the expression of anti-apoptotic protein, bcl-2, was down-regulated in SBC-3/NEO cells, while that of SBC-3/OPN cells was not altered. In contrast, pro-apoptotic protein, bax, was not altered in both SBC 3/OPN and SBC-3/NEO cells, thus bcl-2/bax ratio was decreased in SBC-3/NEO but not altered in SBC-3/OPN cells. Activation of caspase-3 and caspase-9 was increased in SBC-3/NEO cells, but not in SBC-3/OPN cells. Our results suggest that OPN enhances chemo-resistance of cisplatin in SBC-3 cells by suppressing bcl 2 protein down-regulation, thereby blocking the caspase-9- and caspase-3 dependent cell apoptosis. PMID- 19285750 TI - Genetic variation in telomere maintenance genes, telomere length, and lung cancer susceptibility. AB - Telomeres are responsible for the protection of the chromosome ends and shortened telomere length has been associated with risk of multiple cancers. Genetic variation in telomere-related genes may alter cancer risk associated with telomere length. Using lung cancer cases (n=120) and population-based controls (n=110) from Xuanwei, China, we analyzed telomere length separately and in conjunction with single nucleotide polymorphisms in the telomere maintenance genes POT1, TERT, and TERF2, which we have previously reported were associated with risk of lung cancer in this study. POT1 rs10244817, TERT rs2075786, and TERF2 rs251796 were significantly associated with lung cancer (p(trend)< or =0.05). The shortest tertile of telomere length was not significantly associated with risk of lung cancer (OR=1.58; 95% CI=0.79-3.18) when compared to the longest tertile of telomere length. When stratified by genotype, there was a suggestion of a dose-response relationship between tertiles of telomere length and risk of lung cancer among the POT1 rs10244817 common variant carriers (OR (95% CI)=1.33 (0.47-3.75), 3.30 (1.14-9.56), respectively) but not among variant genotype carriers (p(interaction)=0.05). Our findings provide evidence that telomere length and genetic variation in telomere maintenance genes may be associated with risk of lung cancer susceptibility and warrant replication in larger studies. PMID- 19285751 TI - Stage IIIA-N2 NSCLC: a review of its treatment approaches and future developments. AB - Few issues are as controversial in non-small cell lung cancer as the management of patients with stage IIIA-N2 non-small cell lung cancer. This manuscript reviews the reasons and the biases inherent to this controversy and discusses the different treatment approaches with emphasis on survival as evidenced by meta analyses and large randomised clinical trials. Prospects on novel treatment modalities and future research opportunities are presented. PMID- 19285752 TI - Epigenetic changes detected in micropropagated hop plants. AB - Micropropagation is a widely used technique in hops (Humulus lupulus L.). However, to the best of our knowledge, the genetic and epigenetic stability of the microplants has never been tested before. In the present study, two hop accessions were established in vitro and micropropagated for 2 years. The genetic and epigenetic stability of the in vitro plants was analyzed with several molecular techniques: random amplified DNA polymorphism (RAPD), retrotransposon microsatellite amplified polymorphism (REMAP), and methylation-sensitive amplification polymorphism (MSAP). No genetic variation among control and treated plants was found, even after 12 cycles of micropropagation. Epigenetic variation was detected, first, when field and in vitro samples were compared. Nearly a 30% of the detected fragments presented the same pattern of alterations in all the vitroplants. Second, lower levels of epigenetic variation were detected among plants from the different subcultures. Part of this detected variation seemed to be accumulated along the 12 sequential subcultures tested. PMID- 19285753 TI - Evidence-based Emergency Medicine/Critically Appraised Topic. The efficacy of recombinant activated factor VII in severe trauma. AB - STUDY OBJECTIVE: The use of recombinant activated factor VII (rFVIIa) in severe trauma is controversial. This evidence-based emergency medicine review evaluates the existing evidence about the efficacy and safety of rFVIIa for the management of severe trauma. METHODS: We searched MEDLINE, EMBASE, the Cochrane Library, and other databases. We limited our review to prospective, controlled trials that involved the therapeutic use of rFVIIa in the emergency department phase of care. We included studies with blunt and penetrating severe trauma. The primary outcome measure of interest was mortality. Secondary patient-important outcome measures included neurologic outcome, delayed surgical intervention, and adverse effects. Standard criteria were used to evaluate the quality of published trials. RESULTS: One randomized, blinded trial met the inclusion criteria. There was no significant difference in mortality or adverse effects between rFVIIa and placebo. Our other selected secondary outcome measures of interest were not reported. CONCLUSION: Existing evidence suggests that there is no significant difference in mortality between rFVIIa and placebo. Further research is needed to better understand the efficacy and safety of rFVIIa in patients with severe trauma. PMID- 19285754 TI - APIC's role in emergency management: proceedings of the 2008 APIC Emergency Preparedness Mini-Summit. PMID- 19285755 TI - The effects of apolipoprotein E on non-impaired cognitive functioning: a meta analysis. AB - Nearly twice as many participants are represented in the current literature than were available at the time of the last major meta-analytic neurocognitive examination of apolipoprotein E (ApoE) epsilon allele combinations [Small, B.J., Rosnick, C.B., Fratiglioni, L., Backman, L., 2004. Apolipoprotein E and cognitive performance: a meta-analysis. Psychol. Aging 19, 592-600]. The meta-analysis in the current study sought to specifically examine (1) small effects and (2) possible moderating variables associated with ApoE allele combinations that may have been undiscoverable in previous examinations of smaller data sets. A total of 77 studies, representing 40,942 cognitively healthy adults were identified for inclusion in the current meta-analysis (random effects design). Results were congruent with the previous meta-analytic findings indicating that carriers of ApoE allele 4 (E4) perform significantly worse on measures of episodic memory, executive functioning, and overall global cognitive ability. In addition, the current analysis revealed a small effect suggesting that ApoE allele 4 adversely impacts perceptual speed. In contrast to earlier studies, the results also indicate that increases in age result in significantly larger differences between ApoE E4 carriers and ApoE non-E4 carriers on measures of episodic memory and global cognitive ability. ApoE E4 exerts broad, but specific, adverse small effects on a range of neurocognitive functions in cognitively healthy adults. PMID- 19285756 TI - Synthesis and antinociceptive activity of capsinoid derivatives. AB - According to the data of structural identification, six capsinoids or their derivatives were successfully synthesized to test for their analgesic activity. Three of them were capsinoids with different acyl chain compared with capsaicin after substitution of ester for amide at C(1) position. The other three could be described as capsinoid derivatives with different alkoxy chain, compared with capsaicin after substitution of ester for amide at C(1) position and alkoxy for hydroxy at C(4) position and synthesis of them was reported first. Compared with capsaicin, experiment results about pungency showed that capsinoids and their derivatives synthesized were all no or only slight pungent; that is, capsinoid derivates synthesized still have the same advantage of nonpungency with capsinoid. Relation between analgesic activity and molecular structure of compounds synthesized was also reported first, which would facilitate finding capsinoid derivatives owning excellent analgesic activity. The experiment results about analgesic activity showed that capsinoids displayed moderate analgesia effect and their antinociceptive activity decreased with the elongation of acyl chain at C(1) position; that antinociceptive activities of capsinoid derivatives synthesized were much stronger not only than those of indomethacin but also than those of their precursor (vanillyl decanoate), which increased with elongation of alkoxyl chain at C(4) position. Especially 4-hexyloxyl-3-methoxybenzyl decanoate showed the best antinociceptive activity in synthesized compounds, which was 9 fold higher than its precursor (vanillyl decanoate) and 6-fold higher than that of indomethacin. PMID- 19285757 TI - Synthesis and anti-HSV-1 evaluation of some pyrazoles and fused pyrazolopyrimidines. AB - 5-Amino-1-substituted-1H-pyrazole-4-carbonitrile derivative 1 was used as a precursor for preparation of some novel substituted pyrazole and pyrazolo[3,4 d]pyrimidine derivatives 2-10. Furthermore, the preparation of sugar hydrazone derivatives 11a,b, 12a,b and their annelated C-nucleosides 13a,b was described. Some of the prepared products revealed promising antiviral activity against herpes simplex virus type-1 (HSV-1) in comparison to Acyclovir as a control. PMID- 19285758 TI - Synthesis of a series of N6-substituted adenosines with activity against trypanosomatid parasites. AB - The involvement of purine salvage in the accumulation of current trypanocidal drugs is important for the treatment of African sleeping sickness. The substrate specificity of essential nucleoside transporters is therefore of physiological and pharmacological interest. With the intention to contribute to the knowledge in the field, a series of 16 adenosine derivatives with substituents in N(6) position were prepared in order to evaluate their potential to inhibit Trypanosoma brucei spp. in vitro. An unmodified ribose moiety was selected to conserve key molecular recognition motifs of the arsenal of integral membrane proteins expressed in large numbers on the protozoan plasma membrane. Two of the new compounds prepared using a polymer-assisted acylation protocol showed antitrypanosomal activities in the single digit micromolar concentration range. PMID- 19285759 TI - Berbamine derivatives: a novel class of compounds for anti-leukemia activity. AB - Our previous studies showed that the natural compound berbamine, from Chinese herb Berberis amurensis, selectively induces apoptosis of imatinib (IM)-resistant Bcr/Abl-expressing leukemia cells from the K562 cell line and CML patients. Here, a series of new berbamine derivatives were obtained by synthesis. In this series, high to very high activity in vitro has been found. Compounds 2e, 2g, 3f, 3k, 3q and 3u exhibited consistent high anti-tumor activity for imatinib-resistant K562 leukemia cells. Their IC(50) values at 48h were 0.36-0.55 microM, whereas berbamine IC(50) value was 8.9 microM. Cell cycle analysis results showed that compound 3h could reduce G0/G1 cells. In particular, these compounds displayed potent inhibition of the cytoplasm-to-nucleus translocation of NF-kappaB p65 which plays a critical role in the survival of leukemia stem cells. These results suggest that berbamine could be a good starting point for the development of novel lead compounds in the fight against leukemia. PMID- 19285761 TI - An exploration of student nurses' experiences of formative assessment. AB - The idea that formative assessment has the potential to prepare students, not only to succeed in summative assessments during the course, but also in the world beyond the classroom [Melland, H., Volden, C., 1998. Classroom assessment: linking teaching and learning. Journal of Nursing Education 37(6), 275-277] fuelled the desire to explore student nurses experiences of being assessed formatively. Focus group discussion, within a UK Higher Education setting, captured the holistic, dynamic and individual experiences student nurses (n=14) have of formative assessment. Ethical approval was obtained. Findings from three separate focus group discussions indicate that lecturers do not use the term "formative assessment" in their communication with the student nurses; student preparation and effort is greater when assessment is for summative purposes; oral feedback is preferable to written feedback which can, at times, be illegible and utilise unfamiliar vocabulary; lecturer comments are regarded as being more valuable than grades; student nurses are not being prepared for the critical feedback associated with peer review and they may, therefore, be vulnerable to the process and outcome of peer review. Thus, the UK centric focus of this small qualitative research study need not detract from its ability to add to the global knowledge base on formative assessment in nursing. PMID- 19285762 TI - Nutritional support and tumour growth in humans: a narrative review of the literature. AB - BACKGROUND & AIMS: Although there is an experimental evidence that intravenous administration of nutrients may increase tumour growth in animals, data on humans are scanty and sparse. The purpose of this study was to review the literature that has specifically investigated the effects of nutritional support on tumour growth in cancer patients. METHODS: A review of the literature identified 12 suitable papers representing a total of 140 patients receiving nutritional support versus 84 controls. The studies were classified as randomized clinical trials (5), comparative non-randomized clinical trials (3) and trials with patients who were controls for themselves (4). The indicators of tumour growth used in the studies included the DNA index, ornithine decarboxylase activity, flow-cytometric DNA distribution, and the labeling index with tritiated thymidine or bromodeoxyuridine. RESULTS: Increased tumour growth was not observed in control patients without nutritional support but it was reported in 7 out of 12 studies in patients receiving nutritional support. CONCLUSIONS: Providing nutritional support to cancer patients may cause tumours to grow more quickly. However, nutritional support is recommended when nutritional status is so compromised that patients are at high risk for complications or cannot comply with the oncologic therapy as reported in the clinical practice ESPEN guidelines. PMID- 19285766 TI - Surface EMG based muscle fatigue evaluation in biomechanics. AB - In the last three decades it has become quite common to evaluate local muscle fatigue by means of surface electromyographic (sEMG) signal processing. A large number of studies have been performed yielding signal-based quantitative criteria of fatigue in primarily static but also in dynamic tasks. The non-invasive nature of this approach has been particularly appealing in areas like ergonomics and occupational biomechanics, to name just the most prominent ones. However, a correct appreciation of the findings concerned can only be obtained by judging both the scientific value and practical utility of methods while appreciating the corresponding advantages and limitations. The aim of this paper is to serve as a state of the art summary of this issue. The paper gives an overview of classical and modern signal processing methods and techniques from the standpoint of applicability to sEMG signals in fatigue-inducing situations relevant to the broad field of biomechanics. Time domain, frequency domain, time-frequency and time-scale representations, and other methods such as fractal analysis and recurrence quantification analysis are described succinctly and are illustrated with their biomechanical applications, research or clinical alike. Examples from the authors' own work are incorporated where appropriate. The future of this methodology is projected by estimating those methods that have the greatest chance to be routinely used as reliable muscle fatigue measures. PMID- 19285767 TI - ESB Clinical Biomechanics Award 2008: Complete data of total knee replacement loading for level walking and stair climbing measured in vivo with a follow-up of 6-10 months. AB - BACKGROUND: Detailed information about the loading of the knee joint is required for various investigations in total knee replacement. Up to now, gait analysis plus analytical musculo-skeletal models were used to calculate the forces and moments acting in the knee joint. Currently, all experimental and numerical pre clinical tests rely on these indirect measurements which have limitations. The validation of these methods requires in vivo data; therefore, the purpose of this study was to provide in vivo loading data of the knee joint. METHODS: A custom made telemetric tibial tray was used to measure the three forces and three moments acting in the implant. This prosthesis was implanted into two subjects and measurements were obtained for a follow-up of 6 and 10 months, respectively. Subjects performed level walking and going up and down stairs using a self selected comfortable speed. The subjects' activities were captured simultaneously with the load data on a digital video tape. Customized software enabled the display of all information in one video sequence. FINDINGS: The highest mean values of the peak load components from the two subjects were as follows: during level walking the forces were 276%BW (percent body weight) in axial direction, 21%BW (medio-lateral), and 29%BW (antero-posterior). The moments were 1.8%BW*m in the sagittal plane, 4.3%BW*m (frontal plane) and 1.0%BW*m (transversal plane). During stair climbing the axial force increased to 306%BW, while the shear forces changed only slightly. The sagittal plane moment increased to 2.4%BW*m, while the frontal and transversal plane moments decreased slightly. Stair descending produced the highest forces of 352%BW (axial), 35%BW (medio-lateral), and 36%BW (antero-posterior). The sagittal and frontal plane moments increased to 2.8%BW*m and 4.6%BW*m, respectively, while the transversal plane moment changed only slightly. INTERPRETATION: Using the data obtained, mechanical simulators can be programmed according to realistic load profiles. Furthermore, musculo-skeletal models can be validated, which until now often lacked the ability to predict properly the non-sagittal load values, e.g. varus-valgus and internal-external moments. PMID- 19285768 TI - Differences in gait parameters between healthy subjects and persons with moderate and severe knee osteoarthritis: a result of altered walking speed? AB - BACKGROUND: While knee osteoarthritis has been shown to affect a multitude of kinematic, kinetic and temporo-spatial gait parameters, few investigations have examined the effect of increasing levels of radiographic osteoarthritis severity on these gait parameters. Fewer still have investigated the effect of walking speed on gait variables in persons with knee osteoarthritis. The objective of this study was to investigate the influence of walking speed on biomechanical variables associated with joint loading in persons with varying severities of medial compartment knee osteoarthritis. METHODS: Twenty-one persons with moderate osteoarthritis (Kellgren-Lawrence score 2-3) and 13 persons with severe osteoarthritis (Kellgren-Lawrence score of 4) participated. Twenty-two persons without knee pain or radiographic evidence of arthritis comprised a healthy control group. Sagittal plane kinetics, knee adduction moment, sagittal plane knee excursion, ground reaction forces and knee joint reaction forces were calculated from three-dimensional motion analysis at 1.0m/s, self-selected and fastest tolerable walking speeds. Differences were analyzed using multivariate analysis of variance and multivariate analysis of covariance with speed as a covariate. FINDINGS: Persons with knee osteoarthritis showed significantly lower knee and ankle joint moments, ground reaction forces, knee reaction force and knee excursion when walking at freely chosen speeds. When differences in walking speed were accounted for in the analysis, the only difference found at all conditions was decreased knee joint excursion. INTERPRETATION: Compared to a healthy control group, persons with knee OA demonstrate differences in joint kinetics and kinematics. Except for knee excursion, these differences in gait parameters appear to be a result of slower freely chosen walking speeds rather than a result of disease progression. PMID- 19285769 TI - Ozone and Mediterranean ecology: plants, people, problems. PMID- 19285770 TI - Tissue accumulation of aluminium is not a predictor of toxicity in the freshwater snail, Lymnaea stagnalis. AB - The amount of toxic metal accumulated by an organism is often taken as an indicator of potential toxicity. We investigated this relationship in the freshwater snail, Lymnaea stagnalis, exposed to 500 microg l(-1) Al over 30 days, either alone or in the presence of phosphate (500 microg l(-1) P) or a fulvic acid surrogate (FAS; 10 mg l(-1) C). Behavioural activity was assessed and tissue accumulation of Al quantified. Lability of Al within the water column was a good predictor of toxicity. FAS increased both Al lability and behavioural dysfunction, whereas phosphate reduced Al lability, and completely abolished Al induced behavioural toxicity. Tissue accumulation of Al was not linked to toxicity. Higher levels of Al were accumulated in snails exposed to Al + P, compared to those exposed to Al alone, whereas FAS reduced Al accumulation. These findings demonstrate that the degree of tissue accumulation of a metal can be independent of toxicity. PMID- 19285771 TI - Inaccurate perception of asthma symptoms: a cognitive-affective framework and implications for asthma treatment. AB - Inaccurate perception of respiratory symptoms is often found in asthma patients. Typically, patients who inaccurately perceive asthma symptoms are divided into underperceivers and overperceivers. In this paper we point out that this division is problematic. We argue that little evidence exists for a trait-like stability of under- and overperception and that accuracy of respiratory symptom perception is highly variable within persons and strongly influenced by contextual information. Particularly, expectancy and affective cues appear to have a powerful influence on symptom accuracy. Based on these findings and incorporating recent work on associative learning, attention and mental representations in anxiety and symptom perception, we propose a cognitive-affective model of symptom perception in asthma. The model can act as a framework to understand both normal perception as well as under- and overperception of asthma symptoms and can guide the development of affect-related interventions to improve perceptual accuracy, asthma control and quality of life in asthma patients. PMID- 19285772 TI - Face-name repetition priming in semantic dementia: a case report. AB - Repetition priming (RP) has been employed as a measure of implicit processing in patients suffering from a breakdown of semantic memory, as in the case of semantic dementia (SD), a subtype of frontotemporal lobar degeneration (FTLD). Here, we investigated face-name representation in a case of SD using a paradigm of within- and cross-domain repetition priming. Compared to ten healthy participants, SD patient did not show any facilitation when a famous name was primed by its own face (cross-domain) or when the prime was the same proper name (within-domain). Results are discussed within the hypothesis of a degradation of face and name representation, one of the most consistent accounts explaining semantic deficits in SD. PMID- 19285773 TI - A review of recent insights into the role of the sarcoplasmic reticulum and Ca entry in uterine smooth muscle. AB - The uterine sacroplasmic reticulum (SR) takes up and stores calcium [Ca], using an ATPase (SERCA) and the Ca-buffering proteins, calsequestrin and calreticulin. This stored Ca can be released via IP(3)-gated Ca channels. Decreases in luminal Ca concentration [Ca] have been directly measured following agonist stimulation. During spontaneous contractions however, there appears to be no involvement of the SR, as Ca entry and efflux across the plasma membrane account for these phasic contractions. After over-viewing current knowledge concerning SR structure and function, we highlight three areas of research which suggest new ways of looking at the role of the SR in the uterus, although they may be controversial or speculative at the moment. Firstly, we review the evidence for the function, if any, of Ca-induced SR Ca release channels, the ryanodine receptor (RyR) and the lack of Ca sparks (the elemental release events from RyRs), in the uterus. Secondly, we ask does regulation of SERCA by the accessory protein, phospholamban, occur in the uterus and what is the effect of knocking out phospholamban on uterine activity? Thirdly, we address the question of when and how store-operated Ca entry occurs in the myometrium. By analogy with other, usually less excitable tissues, is there a mechanism that links store Ca depletion to plasma membrane Ca entry in smooth muscle cells within intact uterus and is it physiologically relevant and regulated? Are the recently described proteins ORAI and STIM-1 involved in uterine store-operated Ca entry? We end the review by integrating these new insights with previous data to present a new working model of the SR in the uterus. PMID- 19285774 TI - Uterus models for use in virtual reality hysteroscopy simulators. AB - OBJECTIVES: Virtual reality models of human organs are needed in surgery simulators which are developed for educational and training purposes. A simulation can only be useful, however, if the mechanical performance of the system in terms of force-feedback for the user as well as the visual representation is realistic. We therefore aim at developing a mechanical computer model of the organ in question which yields realistic force-deformation behavior under virtual instrument-tissue interactions and which, in particular, runs in real time. STUDY DESIGN: The modeling of the human uterus is described as it is to be implemented in a simulator for minimally invasive gynecological procedures. To this end, anatomical information which was obtained from specially designed computed tomography and magnetic resonance imaging procedures as well as constitutive tissue properties recorded from mechanical testing were used. In order to achieve real-time performance, the combination of mechanically realistic numerical uterus models of various levels of complexity with a statistical deformation approach is suggested. In view of mechanical accuracy of such models, anatomical characteristics including the fiber architecture along with the mechanical deformation properties are outlined. In addition, an approach to make this numerical representation potentially usable in an interactive simulation is discussed. RESULTS AND CONCLUSIONS: The numerical simulation of hydrometra is shown in this communication. The results were validated experimentally. In order to meet the real-time requirements and to accommodate the large biological variability associated with the uterus, a statistical modeling approach is demonstrated to be useful. PMID- 19285775 TI - Multiplex fluorescent PCR for noninvasive prenatal detection of fetal-derived paternally inherited diseases using circulatory fetal DNA in maternal plasma. AB - OBJECTIVE: To develop a fluorescent polymerase chain reaction (PCR) assay for the detection of circulating fetal DNA in maternal plasma and use the established multiplex in noninvasive prenatal genetic diagnosis and its further applications in forensic casework. STUDY DESIGN: The DNA template was extracted from 47 pregnant women and the whole blood samples from the stated biological fathers were used to detect genotype. Using multiplex fluorescent PCR at 16 different polymorphic short tandem repeat (STR) loci, maternal DNA extracted from plasma samples at early pregnancy, medium pregnancy and late pregnancy were used to detect genotype. Their husbands' DNA was also used for fetal genotype ascertainment. RESULTS: Multiplex fluorescent PCR with 16 polymorphic short tandem repeats revealed the presence of fetal DNA in all cases. Every pregnant women/husband pair was informative in at least 3 of 16 loci. The chances of detecting paternally inherited fetal alleles ranged from 66.67 to 94.12%. They are 66.67% in early pregnancy, 85.71% in medium pregnancy and 94.12% in late pregnancy. The accuracy of Multiplex PCR assay to detect fetal DNA was 100%. CONCLUSIONS: Circulating fetal DNA analysis can be used as a possible alternative tool in routine laboratory prenatal diagnosis in the near future; this highly polymorphic STR multiplex has greatly improved the chances of detecting paternally inherited fetal alleles compared with other fetal DNA detection systems that use fetus-derived Y sequences to detect only male fetal DNA in maternal plasma. Our proposed technique can be applied to both female and male fetuses, which provides a sensitive, accurate and efficient method for noninvasive prenatal genetic diagnosis and forensic casework. PMID- 19285776 TI - Tissue mechanics, animal models, and pelvic organ prolapse: a review. AB - Pelvic floor disorders such as pelvic organ prolapse, urinary incontinence, and fecal incontinence affect a large number of women each year. The pelvic floor can be thought of as a biomechanical structure due to the complex interaction between the vagina and its supportive structures that are designed to withstand the downward descent of the pelvic organs in response to increases in abdominal pressure. Although previous work has highlighted the biochemical changes that are associated with specific risk factors (i.e. parity, menopause, and genetics), little work has been done to understand the biomechanical changes that occur within the vagina and its supportive structures to prevent the onset of these pelvic floor disorders. Human studies are often limited due to the challenges of obtaining large tissue samples and ethical concerns. Therefore, it is necessary to investigate the use of animal models and their importance in understanding how different risk factors affect the biomechanical properties of the vagina and its supportive structures. In this review paper, we will discuss the different animal models that have been previously used to characterize the biomechanical properties of the vagina: including non-human primates, rodents, rabbits, and sheep. The anatomy and preliminary biomechanical findings are discussed along with the importance of considering experimental conditions, tissue anisotropy, and viscoelasticity when characterizing the biomechanical properties of vaginal tissue. Although there is not a lot of biomechanics research related to the vagina and pelvic floor, the future is exciting due to the significant potential for scientific findings that will improve our understanding of these conditions and hopefully lead to improvements in the prevention and treatment of pelvic disorders. PMID- 19285777 TI - Assessment of the in vivo biomechanical properties of the human uterine cervix in pregnancy using the aspiration test: a feasibility study. AB - OBJECTIVE: To date no diagnostic tool is yet available to objectively assess the in vivo biomechanical properties of the uterine cervix during gestation. METHODS: We show the first clinical application of an aspiration device to assess the in vivo biomechanical properties of the cervix in pregnancy with the aim to describe the physiological biomechanical changes throughout gestation in order to eventually detect pregnant women at risk for cervical insufficiency (CI). RESULTS: Out of 15 aspiration measurements, 12 produced valid results. The stiffness values were in the range between 0.013 and 0.068 bar/mm. The results showed a good reproducibility of the aspiration test. In our previous test series on non-pregnant cervices our repetitive measurements showed a standard deviation of >20% compared to <+/-10% to our data on pregnant cervices. Stiffness values are decreasing with gestational age which indicates a progressive softening of cervical tissue towards the end of pregnancy. Three pregnant women had two subsequent measurements within a time interval of four weeks. Decreasing stiffness values in the range of 20% were recorded. DISCUSSION: This preliminary study on the clinical practicability of aspiration tests showed promising results in terms of reproducibility (reliability) and clinical use (feasibility). Ongoing studies will provide further insights on its usefulness in clinical practice and in the detection of substantial changes of the cervix in pregnancy indicative for threatened preterm birth or cervical insufficiency. PMID- 19285778 TI - Uterine mechanisms of ipsilateral directed spermatozoa transport: Evidence for a contribution of the utero-ovarian countercurrent system. AB - OBJECTIVE: Spermatozoa transport into uterus and fallopian tubes is directed to the side of the dominant follicle and seems to be controlled by the ipsilateral ovary. The objective of this study was to evaluate the temperature in the fallopian tubes as well as the concentrations of estradiol and progesterone in the utero-ovarian veins draining the ipsilateral ovary and compare these to the contralateral side of the uterus. STUDY DESIGN: A prospective clinical study. SETTING: Academic-assisted reproductive technology program. SUBJECTS: Temperature was measured in both oviducts of 10 patients each in the early phase as well as during the late follicular phase during the course of examination of tubal patency and function. Blood samples of the ovarian veins were obtained during hysterectomy in 10 premenopausal patients with regular menstrual cycles. Five of the women were in the early follicular phase and 5 were in the late follicular phase. RESULTS: Late follicular phase temperature as well as concentrations of estradiol and progesterone were significantly higher in the ipsilateral tube and the utero-ovarian veins draining the ipsilateral ovary as compared to the contralateral side. No such differences were found during the early follicular phase of the cycle. CONCLUSIONS: These data support our view that the uterus and fallopian tubes during the late follicular phase immediately before ovulation are composed of two functional units with different functional properties acting as a peristaltic pump resulting in increased transport of spermatozoa into the oviduct ipsilateral to the ovary bearing the dominant follicle and that this effect is mediated in part by the utero-ovarian countercurrent system. PMID- 19285779 TI - Changes in haemodynamic and autonomous nervous system parameters measured non invasively throughout normal pregnancy. AB - OBJECTIVE: To examine non-invasively haemodynamic and autonomous parameters throughout normal pregnancy. STUDY DESIGN: We used the Task Force Monitor 3040i system to retrieve, record, and calculate haemodynamic as well as autonomous parameters. 20 healthy women were included and scheduled for longitudinal examinations throughout normal pregnancy. Heart rate (HR), blood pressure (BP), stroke volume (SV), cardiac output (CO), systemic vascular resistance (SVR), heart rate variability (HRV), blood pressure variability (BPV), and baroreceptor sensitivity (BRS) were measured. Measurements were performed at gestational week 10(+0)-13(+6), 15(+0)-18(+6), 20(+0)-22(+6), and >30(+0). RESULTS: HR increased during gestation showing a significant increase at III versus I trimester (74 bpm vs. 88 bpm, P<.05). Mean arterial pressure remained stable until III trimester, when a significant increase compared to I trimester could be noted (78 mm Hg vs. 86 mm Hg, P<.05). SV and CO remained relatively stable in I and II trimester, and in III trimester significant decreases were observed. In contrast, SVR increased significantly at III trimester (P<.001). Whereas HRV and BPV did not change at different gestational ages, BRS was significantly lower in III trimester compared to I trimester values (P<.05). CONCLUSION: The non-invasive determination of cardiovascular and autonomous parameters throughout pregnancy is possible and the results of this pilot study can serve as basic parameters for classifying and assessing cardiovascular and autonomous changes in pathological conditions in pregnancy such as hypertensive disorders. PMID- 19285780 TI - Copper and zinc bioavailabilities to ryegrass (Lolium perenne L.) and subterranean clover (Trifolium subterraneum L.) grown in biosolid treated Chilean soils. AB - The purpose of this study was assessing Cu and Zn availabilities in soils amended with a biosolid through the determination of their sequentially extracted chemical forms and their relationship with the contents of these metals in ryegrass (Lolium perenne L.) and subterranean clover (Trifolium subterraneum L.) plant tissues cultivated in a greenhouse using four soils classified as Aquic Xerochrepts and Ultic Haploxeralfs representatives of potential areas for biosolids application in the central zone of Chile. The soils were treated with sewage sludge at a rate of 0 and 30 Mg ha(-1). The greenhouse experiment was carried out through a completely randomized block design in a 2 x 4 (biosolid ratexsoil) arrangement, considering three repetitions per treatment. The soils used in the greenhouse experiment before and after cultivation, were sequentially extracted with specific reagents and conditions in order to obtain the following fractions: exchangeable, sodium acetate-soluble, soluble in moderately reducing condition, K(4)P(2)O(7)-soluble, soluble in reducing condition, and soluble in strongly acid and oxidizing condition. It was established that Cu and Zn were predominantly found in soils in less available forms, associated to organic matter, oxides and clay minerals. Zinc concentration in ryegrass plants was higher than that found in subterranean clover plants in biosolid-amended soils. Zinc contents in ryegrass shoot and root correlated with the exchangeable, bound to-carbonate, and bound-to-FeOx metal forms in control soil. Copper and Zn bioavailabilities were estimated through satisfactorily fitted multiple linear regression models, with determination coefficients from 0.77 to 0.99, which showed a positive contribution of the labile metal forms in soils, especially in relation to Zn in both plant species. PMID- 19285781 TI - The reliability of experts' opinions in constructing a composite environmental index: the case of ESI 2005. AB - The complexity of the environment demands a well-constructed composite environmental index (CEI) to provide a useful tool to draw attention to environmental conditions and trends for policy purposes. Among the common difficulties in constructing a proper CEI are uncertainties due to the selection of the most representative underlying variables or indicators. A degree of uncertainty accompanies experts' judgments, and to deal with vague, subjective or inconsistent information, logic other than classic is required. This study analyzes a procedure that uses different experts' opinions in constructing a CEI, with the use of paraconsistent annotated logic. For this, a sensitivity analysis of the Environmental Sustainability Index (ESI 2005) was used as an example to assess the reliability of experts' opinions. The uncertainty due to the disagreement in experts' opinions clearly indicates that the forms we presently use to measure and monitor the actual environment are insufficient, that is, there is a lack of a "science of sustainability". PMID- 19285782 TI - Expression of heat shock proteins after ultrasound exposure in HL-60 cells. AB - One of the important cellular defense mechanisms against stress is the induction of heat shock proteins (HSPs). We have recently demonstrated that a low frequency electromagnetic field is unable to induce the heat shock response (HSR). In the present study, we expanded our investigations to the induction of HSPs, particularly Hsp72, by ultrasound (US). Human promyelocytic leukemia HL-60 cells were exposed in suspension to US at 1, 3 and 10 MHz, as well as combinations of two of these frequencies. The ability of US to induce Hsp72 was tested for different frequencies, intensities and exposure times. In addition, the water bath temperature was varied from 30 to 36 degrees C. The Hsp72 protein expression was determined 4 and 24 h after treatment. We found that the amount of Hsp72 increased with increasing US frequency, reaching its highest level of about 1800%, induced by 10 MHz. After increasing the temperature of the water bath, the amount of Hsp72 in the treated cells was also increased, whereas no induction was observed at 30 degrees C. For all treatment conditions, ultrasound of 1 MHz was unable to significantly induce Hsp72. At 10 MHz, the exposure time was varied from 0 to 20 min. We found that the induction of Hsp72 took place after 5 min of exposure. For a fixed level of absorbed US energy, the continuous regime, as well as a pulsation of 1:2 (5 ms on and 5 ms off) induced the same Hsp72 level. Pulsation of 1:5 (2 ms on and 8 ms off) and 1:10 (1 ms on and 9 ms off) did not show any effect. A single sonication of 20 min, as well as a fractionated sonication of two 10 min exposures induced the same level of Hsp72, whereas four exposures of 5 min reduced the Hsp72 level. At the optimum exposure conditions (10 MHz, 10 min), the concentration of other HSPs was also determined. Hsp27 showed no effect but Hsp32, Hsp40 and Hsp72 were induced. Taken together, these results suggest a synergistic interaction between heat and US. PMID- 19285783 TI - Microbubble stability is a major determinant of the efficiency of ultrasound and microbubble mediated in vivo gene transfer. AB - In the search for an efficient nonviral gene therapy approach for the treatment of genetic disorders of cardiac and skeletal muscle such as Duchenne muscular dystrophy, ultrasound in combination with contrast enhancing microbubbles has emerged as a promising tool for safe and site-specific enhancement of gene delivery. Indeed, microbubble-enhanced gene transfer (MBGT) has been investigated for a wide variety of target sites using both reporter and therapeutic genes. Although a range of different microbubbles have been used for MBGT studies, comparison of their efficiencies is difficult because microbubble concentration and the ultrasound settings used for the application vary considerably. Only two studies to date have attempted a direct comparison of commercially available microbubbles, and both concluded that not all microbubbles show the same efficiencies with MBGT. Thus far, the reason for this is unclear. Here, the efficiency of three commercially available microbubbles--Optison, SonoVue and Sonazoid--was analyzed to understand the microbubble properties that are important for their function as an effective enhancer for gene transfer in vivo. In this study, plasmid DNA or antisense oligonucleotides were delivered by systemic injection with MBGT, focused on the heart. Gene delivery to the heart with equalized concentrations of the three microbubbles showed that Optison and Sonazoid are more efficient in MBGT compared with SonoVue, which showed the weakest gene transfer to the myocardium. Investigations into the properties of these microbubbles showed that size and shell composition did not directly influence MBGT, whereas the microbubbles with increased stability in an ultrasound field showed better MBGT results than those degrading faster. Moreover, the microbubble concentration used for MBGT was also found to be an important factor influencing the efficiency of MBGT. In conclusion, the stability of a microbubble was shown to be a major influential factor for its performance in MBGT, as is the concentration of the microbubbles used. These findings emphasize the importance of detailed investigations into the properties of microbubbles to allow the production of a microbubble specifically designed for optimum performance with MBGT. PMID- 19285784 TI - Blood clot disruption in vitro using shockwaves delivered by an extracorporeal generator after pre-exposure to lytic agent. AB - The standard methods for recanalyzing thrombosed vessels are vascular stenting or administration of thrombolytic drugs. However, these methods suffer from uncertain success rate and side-effects. Therefore, minimally-invasive ultrasound methods have been investigated. In this article, we propose to use shockwaves after pre-exposure to fibrinolytic agent for disrupting thrombus. Shockwaves were delivered by an extracorporeal piezocomposite generator (120 mm in diameter, focused at 97 mm, pulse length = 1.4 micros). In vitro blood clots, made from human blood, were placed at the focal point of the generator. The clots were exposed to shockwaves either with or without prior immersion in a solution of streptokinase. The percentage of lysed clot was determined by weighing the clot before and after treatment. The proportion of lysed clot increased with the pressure at the focus and with the number of shocks. A mean clot reduction of 91% was obtained for 42 MPa in 4-min treatment duration only, without using streptokinase. For a treatment of 2 min at 29 MPa, the clot reduction increased significantly (p < 0.01) from 47% without streptokinase to 82% when streptokinase was used prior to shockwaves. These results also showed no significant damage to streptokinase due to exposure to shockwaves. This study suggests that extracorporeal shockwaves combined with streptokinase is a promising pharmaco mechanical method for treating occlusive thrombus, and should be confirmed by in vivo trials. Additional studies must also be conducted with other fibrinolytic agents, whose abilities to penetrate clots are different. PMID- 19285785 TI - Measurement of the intimal thickness of the carotid artery: comparison between 40 MHz ultrasound and histology in rats. AB - Common carotid artery intimal thickness (IT) has been shown to be as useful as a measurement of the whole layer of intima-media thickness (IMT) for evaluating the early phase of atherosclerosis. The aim of this study was to elucidate the relationship between high echogenic intimal thickening (HEIT), which was measured using a 40 MHz ultrasound biomicroscope (UBM), and the histologically determined IT in rat carotid arteries. HEIT was estimated in 10 Wistar-Kyoto rats (group I), 15 spontaneous hypertensive rats (SHR) fed a standard diet (group II) and 10 SHR fed a high-fat diet (group III). IT and IMT measurements were determined in Masson trichrome-stained tissues and were compared with the HEIT and IMT evaluated using the UBM. In group I, the HEIT and the IT were 33 +/- 4 microm and 12 +/- 1 mirom, respectively. In group II, the HEIT and the IT were 68 +/- 8 microm and 16 +/- 2 microm, respectively. In group III, the HEIT and the IT were 65 +/- 26 microm and 33 +/- 14 microm, respectively. In SHR, the HEIT and the mean IMT measured with echography were significantly correlated with the IT and the IMT that were determined by histologic measurement (r = 0.60, p = 0.003 and r = 0.53, p = 0.01, respectively). Moreover, HEIT may be associated with the intimal pathology and atherosclerotic burden. The HEIT measurement is a noninvasive method that may be used to assess atherosclerosis in humans. PMID- 19285786 TI - Editorial comment on: A new generation of optical diagnostics for bladder cancer: technology, diagnostic accuracy, and future applications. PMID- 19285787 TI - A new generation of optical diagnostics for bladder cancer: technology, diagnostic accuracy, and future applications. AB - CONTEXT: New developments in optical diagnostics have a potential for less invasive and improved detection of bladder cancer. OBJECTIVE: To provide an overview of the technology and diagnostic yield of recently developed optical diagnostics for bladder cancer and to outline their potential future applications. EVIDENCE ACQUISITION: A PubMed literature search was performed, and papers on Raman spectroscopy (RS), optical coherence tomography (OCT), photodynamic diagnosis (PDD) and narrow-band imaging (NBI) regarding bladder cancer were reviewed. Technology, clinical evidence, and future applications of the techniques are discussed. EVIDENCE SYNTHESIS: With RS, the molecular components of tissue can be measured objectively in qualitative and quantitative ways. The first studies demonstrating human in vivo applicability are still awaited. OCT produces high-resolution, cross-sectional images of tissue, comparable with histopathology, and provides information about depth of tumour growth. The first in vivo studies of OCT demonstrated promising diagnostic accuracy. RS and OCT are not suitable for scanning the entire bladder. PDD is a technique using fluorescence to indicate pathologic tissue. Several studies have shown that PDD increases the detection rate of bladder tumours and improves resection, resulting in fewer early recurrences. The relatively low specificity of PDD remains a problem. NBI enhances contrast of mucosal surface and microvascular structures. The NBI technique has clear advantages over PDD, and the two studies published to date have shown promising preliminary results. PDD and NBI do not contribute to histopathologic diagnosis. CONCLUSIONS: RS and OCT aim at providing a real-time, minimally invasive, objective prediction of histopathologic diagnosis, while PDD and NBI aim at improving visualisation of bladder tumours. For RS, OCT, and NBI, more research has to be conducted before these techniques can be implemented in the management of bladder cancer. All techniques might be of value in specific clinical scenarios. PMID- 19285788 TI - The clinical effectiveness of retropubic (IVS-02) and transobturator (IVS-04) midurethral slings: randomized trial. AB - BACKGROUND: Few series comparing the clinical efficacy of retropubic slings versus transobturator slings for the treatment of female stress urinary incontinence (SUI) are available. OBJECTIVE: To compare clinical efficacy of retropubic tape operations and transobturator suburethral tape operations for the surgical treatment of female SUI. DESIGN, SETTING, AND PARTICIPANTS: From January 2003 to December 2005, 611 patients underwent clinical and urodynamic evaluation before surgical treatment for SUI. Patients with advanced urogenital prolapse (pelvic organ prolapse-quantification scale [POP-Q] scale grade >1) were excluded, and 537 patients were included in this study. After 18 mo, 398 women were available for follow-up efficacy evaluation at a tertiary academic center. INTERVENTION: All patients underwent either a retropubic sling procedure or a transobturator sling procedure. Patients were randomly allocated into two study groups at a ratio of 1:1. MEASUREMENTS: After 18 mo all enrolled patients were clinically checked for clinical efficacy of both procedures. RESULTS AND LIMITATIONS: Demographic and urodynamic parameters of patients were similar in both groups. No bladder injury occurred in the transobturator sling group (IVS 04), whereas 13 intraoperational bladder perforations (6.5%) occurred in the retropubic sling group (IVS-02) (p<0.001). The tape erosion rate was <2.5% in both groups (p=0.7). After 18 mo, 398 patients (201 in the IVS-02 group and 197 in the IVS-04 group) were evaluated in terms of clinical efficacy of the procedures. We found out that there was no statistically significant difference in clinical efficacy between these two procedures (chi(2)=1.88, p=0.39). In the IVS-02 group, 75.1% of patients (n=151) remained dry (cured), 16.9% of patients (n=34) reported significant improvement, and 8.0% of patients (n=16) were considered as failures. In the IVS-04 group, 74.1% of patients (n=146) remained dry, 14.2% of patients (n=28) reported significant improvement, and 11.7% (n=23) were considered as failures. CONCLUSIONS: Based on an 18-mo follow-up, the efficacies of both techniques are comparable; however, the retropubic route appears to be more efficient in the intrinsic sphincter deficiency (ISD) group. PMID- 19285789 TI - Editorial comment on: A new generation of optical diagnostics for bladder cancer: technology, diagnostic accuracy, and future applications. PMID- 19285790 TI - Adult medulloblastoma and the "dural-tail" sign: rare mimic of a posterior petrous meningioma. AB - The authors describe clinical and radiological features of a 32-year-old male patient with a posterior fossa lesion. The radiological characteristics resembled an extra-axial lesion; a meningioma, with attachment to the tentorium and petrous bone. The histological diagnosis was classical medulloblastoma. We review literature of this atypical presentation of medulloblastoma and "dural-tail" sign, which can be associated with other benign or malignant lesions. The role of magnetic resonance spectroscopy in the diagnosis of primitive neuroectodermal tumors is discussed. PMID- 19285791 TI - Naive human umbilical cord matrix derived stem cells significantly attenuate growth of human breast cancer cells in vitro and in vivo. AB - The effect of un-engineered (naive) human umbilical cord matrix stem cells (hUCMSC) on the metastatic growth of MDA 231 xenografts in SCID mouse lung was examined. Three weekly IV injections of 5x10(5) hUCMSC significantly attenuated MDA 231 tumor growth as compared to the saline-injected control. IV injected hUCMSC were detected only within tumors or in close proximity to the tumors. This in vivo result was corroborated by multiple in vitro studies such as colony assay in soft agar and [(3)H]-thymidine uptake. These results suggest that naive hUCMSC may be a useful tool for cancer cytotherapy. PMID- 19285792 TI - Photodegradation of bisphenol-A in a batch TiO2 suspension reactor. AB - In this work, the photocatalytic behaviors of bisphenol-A (BPA), which has been listed as one of endocrine disrupting chemicals, were carried out in a batch TiO(2) suspension reactor. The photodegradation efficiency has been investigated under the controlled process parameters including initial BPA concentration (i.e., 1-50 mg L(-1)), TiO(2) dosage (i.e., 5-600 mg/200 cm(3)), initial pH (i.e., 3-11), and temperature (i.e., 10-70 degrees C). It was found that the optimal conditions in the photoreaction process could be coped with at initial BPA concentration=20 mg L(-1), TiO(2) dosage=0.5 g L(-1) (100mg/200 cm(3)), initial pH=7.0, and temperature=25 degrees C. According to the Langmuir Hinshelwood model, the results showed that the photodegradation kinetics for the destruction of BPA in water also followed the first-order model well. The apparent first-order reaction constants (k(obs)), thus obtained from the fittings of the model, were in line with the destruction-removal efficiencies of BPA in all the photocatalytic experiments. Based on the intermediate products identified in the study, the possible mechanisms for the photodegradation of BPA in water were also proposed in the present study. PMID- 19285793 TI - Adsorption of Eu(III) onto TiO2: effect of pH, concentration, ionic strength and soil fulvic acid. AB - The effects of pH, initial Eu(III) concentration, ionic strength and fulvic acid (FA) on the adsorption of Eu(III) on TiO(2) are investigated by using batch techniques. The results indicate that the presence of FA strongly enhances the adsorption of Eu(III) on TiO(2) at low pH values. Besides, the adsorption of Eu(III) on TiO(2) is significantly dependent on pH values and independent of ionic strength. The adsorption of Eu(III) on TiO(2) is attributed to inner-sphere surface complexation. The diffuse layer model (DLM) is applied to simulate the adsorption data, and fits the experimental data well with the aid of FITEQL 3.2. X-ray photoelectron spectroscopy (XPS) is performed to study the species of Eu(III) adsorbed on the surfaces of TiO(2)/FA-TiO(2) hybrids at a molecular level, which suggest that FA act as "bridge" between Eu(III) and TiO(2) particles to enhance the ability to adsorb Eu(III) in solution. PMID- 19285794 TI - Effect of recycling flux on performance and characteristics of activated sludge hydrolytic-aerobic recycling process in degradation of 2,4-dichlorophenol. AB - The degradation of a model molecule, 2,4-dichlorophenol (2,4-DCP), was studied using an activated sludge hydrolytic-aerobic recycling process (HARP). 2,4-DCP and chemical oxygen demand (COD) total removal efficiency in the recycling process was 98% and 96% at the recycling flux of 15 mL/min after 24h degradation, respectively. With the recycling flux increasing, the peak values of the concentration of volatile fatty acid (VFA) declined. Polysaccharide and protein contents in EPS were dramatically increased as recycling flux increasing from 5 to 15 mL/min in the HARP. There is obviously fit to the linear correlation between the PN/PS ratios and recycling flux. The zeta potentials decreased with recycling flux increasing. As the recycling flux increasing at a certain degree, the increase in polysaccharide and protein contents of EPS could more favor the stability of the HARP. PMID- 19285795 TI - Oxidation of polyvinyl alcohol by persulfate activated with heat, Fe2+, and zero valent iron. AB - The oxidation of polyvinyl alcohol (PVA) by persulfate (S(2)O(8)(2-)) activated with heat, Fe(2+), and zero-valent iron (Fe(0)) was investigated via batch experiments. It was hypothesized that elevated temperature and the addition of Fe(2+) or Fe(0) into a persulfate-water system could enhance the oxidation of PVA by activated persulfate. Increasing the temperature from 20 to 60 degrees C or 80 degrees C accelerated the oxidation rate of PVA, which achieved complete oxidation in 30 and 10 min, respectively. At 20 degrees C, the addition of Fe(2+) or Fe(0) to the persulfate-water system significantly enhanced the oxidation of PVA. The optimal persulfate-to-Fe(2+) or Fe(0) molar ratio was found to be 1:1. Complete oxidation of PVA was obtained by Fe(0)-activated persulfate in 2h. Synergistic activation of persulfate by heat and Fe(2+) or Fe(0) was also shown to enhance the oxidation of PVA in the persulfate-water system. By using GC-MS analysis, an oxidation product of PVA was identified as vinyl acetic acid (C(4)H(6)O(2)), which is readily biodegradable. Our results suggest that the oxidative treatment of PVA by activated persulfate is a viable option for the pretreatment of PVA-laden wastewater to enhance its biodegradability. PMID- 19285796 TI - Thermophilic biofiltration of H2S and isolation of a thermophilic and heterotrophic H2S-degrading bacterium, Bacillus sp. TSO3. AB - Thermophilic biofiltration of H(2)S-containing gas was studied at 60 degrees C using polyurethane (PU) cubes and as a packing material and compost as a source of thermophilic microorganisms. The performance of biofilter was enhanced by pH control and addition of yeast extract (YE). With YE supplement and pH control, H(2)S removal efficiency remained above 95% up to an inlet concentration of 950 ppmv at a space velocity (SV) of 50h(-1) (residence time=1.2 min). H(2)S removal efficiency strongly correlated with the inverse of H(2)S inlet concentrations and gas flow rates. Thermophilic, sulfur-oxidizing bacteria, TSO3, were isolated from the biofilter and identified as Bacillus sp., which had high similarity value (99%) with Bacillus thermoleovorans. The isolate TSO3 was able to degrade H(2)S without a lag period at 60 degrees C in liquid cultures as well as in the biofilter. High H(2)S removal efficiencies were sustained with a periodic addition of YE. This study demonstrated that an application of thermophilic microorganism for a treatment of hot gases may be an economically attractive option since expensive pre-cooling of gases to accommodate mesophilic processes is not required. PMID- 19285797 TI - Assessment of odorous VOCs released from a main MSW landfill site in Istanbul Turkey via a modelling approach. AB - An air pollution modeling study was conducted to investigate the odorous effects of volatile organic compounds (VOCs) emissions from a sanitary landfill area on ambient air quality. The atmospheric dispersion of hydrogen sulfide (H(2)S) and 22 VOCs was modeled. Industrial Source Complex v3 Short Term (ISCST3) model was used to estimate hourly concentrations of odorous VOCs over the nearest residential area. Odor thresholds of VOCs of interest were also found in the literature. Results showed that short-term averages of three odorous VOCs, namely ethyl mercaptan, methyl mercaptan and hydrogen sulfide, exceeded their odor thresholds, which are reported to be 0.022, 0.138 and 11.1 microg/m(3), respectively, at several points within the domain. Their highest concentrations within Gokturk County were estimated to be 0.09387 microg/m(3) for ethyl mercaptan, 0.07934 microg/m(3) for methyl mercaptan and 6.315 microg/m(3) for hydrogen sulfide. Short-term model results revealed the occasional odor problems being reported for Gokturk County. Hourly concentrations were used to obtain frequencies of odor episodes in Gokturk County via a probability analysis. The results showed that ethyl mercaptan concentrations did not exceed its odor threshold during more than 8.84% of the time. Similarly, the maximum odor episode frequencies for methyl mercaptan and hydrogen sulfide were 0.98% and 0.34% of the time, respectively. PMID- 19285798 TI - Biosorption of cadmium (II) and lead (II) from aqueous solutions using mushrooms: a comparative study. AB - Sorption capacity of oyster mushroom (Pleurotus platypus), button mushroom (Agaricus bisporus) and milky mushroom (Calocybe indica) were evaluated on biosorption of heavy metals, viz. cadmium (II) and lead (II) from aqueous solutions. The optimum sorption conditions were studied for each metal separately. The desired pH of the aqueous solution was found to be 6.0 for the removal of cadmium (II) and 5.0 for removal of lead (II) for all the mushrooms. The percent removal of both the metals was found to increase with the increase in biosorbent dosage and contact time. The fitness of the biosorption data for Langmuir and Freundlich adsorption models was investigated. It was found that biosorption of cadmium (II) and lead (II) ions onto the biomass of the three mushrooms were better suitable to Langmuir than Freundlich adsorption model. P. platypus showed the highest metal uptake potential for cadmium (q(max) 34.96 mg/g) whereas A. bisporus exhibited maximum potential for lead (q(max) 33.78 mg/g). Milky mushroom showed the lowest metal uptake capacity for both the metals. The present data confirms that mushrooms may be used as efficient biosorbent for the removal of cadmium (II) and lead (II) ions from aqueous solution. PMID- 19285799 TI - Determination of parameters used to prevent ignition of stored materials and to protect against explosions in food industries. AB - There are always risks associated with silos when the stored material has been characterized as prone to self-ignition or explosion. Further research focused on the characterization of agricultural materials stored in silos is needed due to the lack of data found in the literature. The aim of this study was to determine the ignitability and explosive parameters of several agricultural products commonly stored in silos in order to assess the risk of ignition and dust explosion. Minimum Ignition Temperature, with dust forming a cloud and deposited in a layer, Lower Explosive Limit, Minimum Ignition Energy, Maximum Explosion Pressure and Maximum Explosion Pressure Rise were determined for seven agricultural materials: icing sugar, maize, wheat and barley grain dust, alfalfa, bread-making wheat and soybean dust. Following characterization, these were found to be prone to producing self-ignition when stored in silos under certain conditions. PMID- 19285800 TI - Optimal exponential feeding strategy for dual-substrate biostimulation of phenol degradation using Cupriavidus taiwanensis. AB - The exponential feeding strategy (EFS) of dual substrates (i.e., phenol and glycerol) was applied to optimize the overall performance of phenol degradation by Cupriavidus taiwanensis R186. Addition of a second substrate (e.g., glycerol) could stimulate the phenol biodegradation efficiency of strain R186. Hence, a feasible EFS was developed for fed-batch phenol biodegradation using the dual substrate biostimulation technique. The phenol degradation kinetics was well characterized with proposed model and response surface analysis. Our findings quantitatively revealed that glycerol could effectively enhance the phenol degradation performance, as the highest phenol degradation efficiency occurred with the supplementation of 0.8-1.2 g L(-1) of glycerol. The optimal dual substrate EFS was identified via contour analysis and kinetic modeling. With the optimal dual-substrate EFS (i.e., a feeding rate constant (alpha(1) and alpha(2)) of 0.5 and 0.3, respectively), the shortest time (ca. 13.80 h) for phenol degradation was achieved with a specific growth rate of ca. 0.281 h(-1). PMID- 19285801 TI - Study on the influence of moisture content on thermal stability of propellant. AB - This paper studies the influence of moisture content on the thermal stabilities of double-base propellant and multi-nitro ester propellant. The thermal behaviors and chemical kinetic parameters of the above two propellants and their mixtures with water were analyzed by using a CALVET heat flux calorimeter, C80. The thermal decomposition mechanisms of these two propellants with water were conjectured based on the tests. And then, the self-accelerating decomposition temperatures (SADT) of these two propellants and their mixtures with water were calculated and compared according to the kinetic parameters and Semenov model. The results show that the thermal decomposition mechanism of double-base propellant with water may be changing with the varying moisture content by transferring hydrogen proton (H(+)). However, the thermal decomposition mechanism of multi-nitro ester propellant with water may be unchanging due to the excess of formaldehyde (HCHO). Water plays the external physical factor on the thermal decomposition of multi-nitro ester propellant, and it plays both the physical and the chemical factors on the thermal decomposition of double-base propellant. The SADTs of their mixtures with water are much lower than that of pure propellants, and keep decreasing with the increasing of moisture content. PMID- 19285802 TI - Liquid-liquid extraction/separation of platinum(IV) and rhodium(III) from acidic chloride solutions using tri-iso-octylamine. AB - Liquid-liquid extraction/separation of platinum(IV) and rhodium(III) from acidic chloride solutions was carried out using tri-iso-octylamine (Alamine 308) as an extractant diluted in kerosene. The percentage extraction of platinum(IV) and rhodium(III) increased with increase in acid concentration up to 8 mol L(-1). However, at 10 mol L(-1) HCl concentration, the extraction behavior was reversed, indicating the solvation type mechanism during extraction. The quantitative extraction of approximately 98% platinum(IV) and 36% rhodium(III) was achieved with 0.01 mol L(-1) Alamine 308. The highest separation factor (S.F.=184.7) of platinum(IV) and rhodium(III) was achieved with 0.01 mol L(-1) Alamine 308 at 1.0 mol L(-1) of hydrochloric acid concentration. Alkaline metal salts like sodium chloride, sodium nitrate, sodium thiocyanate, lithium chloride, lithium nitrate, potassium chloride and potassium thiocyanate used for the salting-out effect. LiCl proved as best salt for the extraction of platinum(IV). Temperature effect demonstrates that the extraction process is exothermic. Hydrochloric acid and thiourea mixture proved to be better stripping reagents when compared with other mineral acids and bases. PMID- 19285803 TI - Removal of acid orange 7 by guava seed carbon: a four parameter optimization study. AB - The preparation of carbon from waste materials is a recent and economic alternative for the removal of dyes. In this study four samples of carbon were obtained by thermal treatment at 1000 degrees C using as precursor the guava seed with different particle sizes. The Taguchi method was applied as an experimental design to establish the optimum conditions for the removal of acid orange 7 in batch experiments. The chosen experimental factors and their ranges were: pH (2 12), temperature (15-35 degrees C), specific surface area (50-600 m(2)g(-1)) and adsorbent dosage (16-50 mg ml(-1)). The orthogonal array L(9) and the larger the better response category were selected to determine the optimum removal conditions: pH 2, temperature 15 degrees C, S(esp) 600 m(2)g(-1) and dosage 30 mg ml(-1). Under these conditions a total removal of acid orange 7 was achieved. Moreover, the most significant factors were the carbon specific surface area and the pH. The influence of the different factors on the adsorption of acid orange 7 from solution is explained in terms of electrostatic interactions by considering the dye species and the character of the surface. PMID- 19285804 TI - Treatment of door-manufacturing factories wastewaters using CDEO and other AOPs: a comparison. AB - In this work, three advanced oxidation technologies have been studied to improve the quality of the effluents of a physicochemical process and of a combined physicochemical-biological process during the treatment of actual industrial wastes of wooden door-manufacturing factories. From the treatment point of view (neglecting costs), advanced oxidation processes can be successfully used to treat both, coagulated and biologically treated wastes. Conductive-diamond electrochemical oxidation (CDEO) was found to be the more effective technology because it can reduce completely the chemical oxygen demand (COD) (no production of refractory compounds) with a very high current efficiency. However, from the economic viewpoint, the direct treatment of the coagulated wastes can not be recommended because it is very expensive. Only Fenton oxidation or conductive diamond electrochemical oxidation can be cost-efficiently used to refine the quality of the effluent of the biological process. PMID- 19285805 TI - Ablation of estrogen receptor alpha or beta eliminates sex differences in mechanical pain threshold in normal and inflamed mice. AB - We examined nociceptive responses to mechanical stimulation in mice of both sexes lacking the estrogen receptor alpha or beta and in respective wild types under normal conditions, after inflammation of a hindpaw or peripheral nerve injury. In normal wild-type mice, females had significantly lower paw withdrawal threshold than males. There was no significant difference between wild-type mice and knock outs of either estrogen receptor alpha or beta in mechanical response threshold in male mice. However, significantly elevated response threshold was observed in both knock-out female mice, which eliminated sex differences in nociception. After carrageenan-induced inflammation of a hindpaw, all wild-type and knock-out mice exhibited similar local edema with no difference between the sexes. Wild type mice developed hypersensitivity (allodynia) to mechanical stimulation, which was more profound in the females than in males. Again, such sex difference was not observed in the knock-outs of either estrogen receptor. Photochemically induced partial sciatic nerve injury caused similar persistent mechanical hypersensitivity in the wild types and both estrogen receptor knock-outs with no difference between the sexes. These results suggest that the sex difference in basal mechanical pain threshold and inflammatory hypersensitivity is eliminated in mice lacking either the estrogen alpha receptors or beta receptors. However, these receptors do not seem to be directly involved in mediating pain sensitivity in general or in the development of neuropathic pain. It is unclear whether the elimination of sex differences observed in the knock-outs reflects an ongoing effect of estrogen acting through its receptors in females or the developmental changes that predominantly affect females. PMID- 19285806 TI - Expression and activity of glycogen synthase kinase during vitellogenesis and embryogenesis of Rhipicephalus (Boophilus) microplus. AB - Glycogen synthase kinase 3 (GSK-3) is classically described as a key enzyme involved in glycogen metabolism in mammals. GSK-3 belongs to a highly conserved family of serine/threonine protein kinases, whose members are involved in hormonal regulation, nuclear signaling, and cell fate determination in higher eukaryotes. We have cloned and characterized the RmGSK-3 gene from Rhipicephalus (Boophilus) microplus tick embryos. DNA and protein sequence analysis depicted high similarity to the corresponding enzyme, from both vertebrate and invertebrate animals. In addition, the mRNA transcription profile identified during embryogenesis was analyzed. We observed that the RmGSK-3 mRNA rapidly decreases from the 1st to 3rd day of development, and increases from the 3rd to 15th day. After the 15th day of development, we observed a near 50% reduction in RmGSK-3 mRNA transcription in comparison to the 1st day. We detected the GSK 3beta isoform in egg homogenates throughout embryogenesis using Western blot analysis. RmGSK-3 mRNA was present in fat body, midgut and ovary from partially and fully engorged adult female ticks. The highest mRNA level was observed in ovaries from both developmental stages and in first-day eggs. Furthermore, RmGSK 3 activity correlated with glycogen content variation. Finally, kinase activity in egg homogenates was inhibited by the specific inhibitor, SB-216763. These data suggest that RmGSK-3beta may be involved in glycogen metabolism regulation during R. microplus embryogenesis. PMID- 19285807 TI - Dioctophyme renale (Goeze, 1782) in the abdominal cavity of a domestic cat from Brazil. AB - This study reports a case of parasitism by the giant kidney worm, Dioctophyme renale (Goeze, 1782), in the abdominal cavity of a domestic cat from Brazil. A female adult cat presenting prostration, dehydration, physical debility, pronounced jaundice and ascitis, was taken to the Department of Animal Parasitology of the Veterinary Institute of the Universidade Federal Rural do Rio de Janeiro, Brazil. Clinical signs suggested a case of peritonitis. The cat's clinical condition was grave and death occurred within a few days. During necropsy, a brownish-red nematode, 24.9cm long, was found in the abdominal cavity and was identified as a male adult D. renale. This study reports the first confirmed case of dioctophymatosis in the domestic cat. The parasite's aberrant location in the abdominal cavity suggests that the domestic cat is not a suitable host. PMID- 19285808 TI - Genetic diversity of Anaplasma marginale in Argentina. AB - Bovine anaplasmosis caused by Anaplasma marginale is a worldwide major constraint to cattle production. The A. marginale major surface protein 1 alpha (msp1alpha) gene contains a variable number of tandem repeats in the amino terminal region and has been used for the characterization of pathogen genetic diversity. This study reports the first characterization of A. marginale genetic diversity in Argentina based on msp1alpha genotypes and its putative relationship with Rhipicephalus (Boophilus) microplus infestations. Herein, we analyzed whole blood bovine samples from anaplasmosis outbreaks in R. microplus infested (9 samples) and eradicated/free (14 samples) regions. Sequence analysis revealed the existence of 15 different msp1alpha genotypes with 31 different repeat units. Six new repeat sequences were discovered in this study and 13/31 (42%) repeats were unique to Argentinean strains. The analysis of msp1alpha repeat sequences according to R. microplus infestations resulted in three repeat groups: (i) found in tick-infested regions (20 repeats), (ii) found in tick free regions (6 repeats) and (iii) randomly distributed (5 repeats). Moreover, A. marginale msp1alpha genetic diversity was higher in tick-infested regions than in tick free areas. These results, together with previous evidence suggesting that A. marginale msp1alpha repeat units co-evolved with the tick vector, might represent an evidence of the role of tick-mediated transmission for the generation of pathogen genetic diversity. PMID- 19285809 TI - Seroprevalence of Neospora caninum and Toxoplasma gondii antibodies in white tailed deer (Odocoileus virginianus) from Iowa and Minnesota using four serologic tests. AB - The white-tailed deer (Odocoileus virginianus) is considered one of the most important wildlife reservoirs of Neospora caninum and Toxoplasma gondii in the US. Sera from white-tailed deer from Minnesota and Iowa were tested for antibodies to N. caninum by four serologic tests including the indirect fluorescent antibody (IFA) test (cut-off 1:25), Neospora caninum agglutination test (cut-off 1:25), an enzyme-linked immunoabsorbent assay, and Western blot (WB). Sera were also tested for antibodies to T. gondii using the modified agglutination test (cut-off 1:25). Of 62 adult deer from Minnesota antibodies to T. gondii were found in 20 (32.2%), N. caninum in 44 (71%), with dual infections in 18 deer. Of 170 (73 fawns, 9 yearlings, 88 adults) deer from Iowa, T. gondii antibodies were present in 91 (53.5%) with 37.0, 55.6 and 67.0% seropositivity in fawns, yearlings, and adults, respectively. Antibodies to N. caninum were found in 150 of 170 (88.2%) by any of the 3 tests (99 by Western blots, 135 by ELISA, 106 by IFA, and 118 by NAT). Dual infections with T. gondii and N. caninum were detected in 47 deer. Very high (84.9%) seropositivity of N. caninum in fawns suggests high rate of congenital transmission of the parasite. Seropositivity in each test at different titers is discussed. PMID- 19285810 TI - Measuring principal substance of abuse in comorbid patients for clinical research. AB - Few individuals with substance use disorders limit their intake to one substance of abuse; however, many studies focus on a single substance. Unfortunately, the optimal method to determine the principal substance is unclear. In particular, this issue is problematic in patients with co-occurring psychiatric illness, who commonly use multiple substances. Hence we compared three methods for assessing the principal substance of abuse in 150 subjects with bipolar disorder and substance dependence: 1) the Addiction Severity Index interview, 2) a self administered questionnaire, and 3) the most frequently used substance. While most subjects were concordant on the interview and the other two methods, we found substantial disagreement (9.3% between the interview and the questionnaire, and 12.7% between the interview and the most frequently used substance) and partial agreement (14.0%). These findings from a comorbid population demonstrate that different methods to assess principal substance of abuse could lead to different conclusions about treatment outcomes. Hence studies of comorbid patients may benefit from 1) using more than one method to assess principal substance and 2) reporting use of all substances as well as a targeted substance. PMID- 19285811 TI - Comment on "Vitamin D deficiency is the cause of common obesity". PMID- 19285812 TI - Well-differentiated papillary mesothelioma of the tunica vaginalis: a case report with aspirate cytologic, immunohistochemical, and ultrastructural studies. AB - A case of a well-differentiated papillary mesothelioma of the tunica vaginalis testis in a 70-year-old man, who presented with a 1-month history of scrotal swelling with hydrocele, is described. Fine needle aspiration cytology of the hydrocele demonstrated a papillary cluster of uniform round cells with minimal atypia, reminiscent of mesothelial cells. Macroscopically, the lesion was characterized by a solitary, well-circumscribed papillary soft tumor measuring 1.2 x 1.2 x 1.0 cm(3). Histologically, the tumor exhibited papillary or tubulopapillary proliferation of predominantly bland cuboidal cells. No cellular pleomorphism, mitotic figures, necrosis, or stromal invasion was noted. Immunohistochemically, the tumor was positive for CAM5.2, epithelial membrane antigen, vimentin, calretinin, and D2-40, but negative for carcinoembryonic antigen. Ki-67 index was less than 1%. Ultrastructurally, the tumor cells showed numerous microvilli, mitochondria, rough endoplasmic reticulum, microfilaments, and desmosomes. The patient is alive with no evidence of disease at 18 months after right radical orchiectomy. Ki67 may be potentially a good maker for differentiating low-grade from high-grade mesothelioma. PMID- 19285813 TI - Exercise to reduce vasomotor and other menopausal symptoms: a review. AB - Many women are reluctant to consider HRT as a therapeutic option for menopausal symptoms and are keen to use non-pharmacological treatments. Evidence from randomised controlled trials (RCTs) concerning the effects of aerobic exercise on vasomotor and other menopausal symptoms is limited but what evidence we do have suggests that aerobic exercise can improve psychological health and quality of life in vasomotor symptomatic women. In addition, several RCTs of middle aged/menopausal-aged women have found that aerobic exercise can invoke significant improvements in several common menopause-related symptoms (e.g. mood, health-related QoL and insomnia), relative to non-exercise comparison groups. There is some evidence that alternative forms of low intensity exercise such as yoga are beneficial in reducing vasomotor symptoms and improving psychological well-being in menopausal women. Collectively, these RCTs highlight the broader potential that exercise could have for women during the menopause transition. Whilst both the Royal College of Obstetricians and Gynaecologists in the UK and the North American Menopause Society have recommended that women be advised to consider aerobic exercise as a treatment for vasomotor menopausal symptoms, to make any evidence-based conclusions regarding the effectiveness of exercise in managing these symptoms, more high quality research is needed. PMID- 19285814 TI - Endometrial hyperplasia and the risk of progression to carcinoma. AB - The primary presenting symptom of endometrial neoplasia is abnormal uterine bleeding, which typically prompts an endometrial biopsy to rule out carcinoma. Approximately 70% of women with abnormal uterine bleeding are diagnosed with benign findings and 15% are diagnosed with carcinoma. The remaining 15% receive a diagnosis of endometrial hyperplasia (EH), which includes a broad range of lesions, from mild, reversible proliferations to the immediate precursors of carcinoma. The widely used World Health Organization (WHO) system classifies EH according to four combinations of glandular crowding and nuclear atypia: simple (SH), complex (CH), simple atypical (SAH), or complex atypical hyperplasia (CAH), although the two forms of atypical hyperplasia (AH) are often collapsed into one category. Diagnoses of EH raise three issues. First, the low interobserver reproducibility-less than 50% in almost all studies-hinders the ability of WHO based classification to effectively guide clinical management. Second, approximately 50% of women diagnosed with AH have concurrent carcinoma. Not surprisingly, most women with AH undergo hysterectomy as primary treatment, but non-surgical management can be effective. Third, data on progression risks for women with EH who retain their uterus are extremely limited. Emerging data indicate the long-term risk among women with SH or CH is less than 5%, but the risk among women with AH is approximately 30%. These data highlight priority areas for future research, such as increasing the diagnostic reproducibility of EH, improving the discrimination between AH and carcinoma, and identifying biomarkers to stratify risks or serve as indicators of response to clinical treatment. PMID- 19285815 TI - The genetic message of a sudden, unexpected death due to thoracic aortic dissection. AB - Thoracic aortic aneurysms are associated with sudden, unexpected death due to dissection and/or rupture. In such cases, the latent, preceding state of aortic dilatation has often gone undiagnosed. As a consequence of the sudden unresolved death, medico-legal autopsy requested by a public prosecutor will be the consequence to establish the cause and manner of death. Usually, autopsy records do not include relevant information for differential diagnosis of heritable syndromic and non-syndromic diseases associated with thoracic aortic aneurysms/dissections (TAAD), including e.g. Marfan syndrome, Loeys-Dietz syndrome, and isolated thoracic aortic aneurysms/dissection. However, for at-risk relatives of the deceased, it could be of great benefit to be alerted to the potential heritable aetiology, because early diagnosis of the latent stage of the disease would allow preventive management. Such attempts, including recommendations to seek genetic counselling, are nevertheless rarely made in the context of medico-legal autopsies, in which primarily the legal aspects are considered. We report here on three cases to underline the practical relevance of (i) documentation of relevant information for differential diagnosis of TAAD associated disorders, (ii) storage of unfixed tissue samples for subsequent molecular genetic testing, and most importantly (iii) the information of relatives at risk. In view of the general ethical principal of nonmaleficience, direct or indirect contact with family members of victims of possible heritable forms of TAAD should be established as a standard of care, also in the medico legal setting. PMID- 19285816 TI - Evaluation of tinnitus patients with normal hearing sensitivity using TEOAEs and TEN test. AB - OBJECTIVES: This study was designed to investigate the possibility of underlying cochlear damage whether outer hair cells (OHCs) or inner hair cells (IHCs) in tinnitus suffering patients with normal hearing sensitivity, using transient evoked otoacoustic emission (TEOAEs) and threshold equalizing noise (TEN) test, if any. METHODS: Twenty patients suffering from unilateral tinnitus with normal hearing sensitivity participated in this study. Their other ear acted as control ears. They were subjected to full history taking, otoscopy, basic audiologic evaluation, TEOAEs and TEN test. RESULTS: TEOAEs were abnormal in 85% of the tinnitus ears compared to 20% in control ears; this difference was statistically significant. The abnormal TEOAEs frequency bands in the tinnitus ears were statistically significant above 2000 Hz when compared to the control ears and were more common for the 4000 and 5000 Hz. This suggests that OHCs dysfunction may be important in the generation of tinnitus. TEN test demonstrated dead regions in the cochlea in 15% of the tinnitus ears only. This might be attributed to increased resistance of IHCs to damage compared to OHCs vulnerability. The affected frequency location was at 500 Hz in 5%, 3000 and 4000 Hz in 10% of tinnitus ears. CONCLUSION: This work has shown a higher prevalence of OAE abnormalities in tinnitus patients with normal hearing in contrast to TEN test denoting the more vulnerability of OHCs to damage. PMID- 19285817 TI - [Awareness and adherence to Pertussis vaccination guidelines by occupational medicine physicians in Paris healthcare institution]. AB - OBJECTIVE: A questionnaire was used on 44 public and private hospital physicians in Paris to evaluate their knowledge of and adherence to Vaccination Guidelines, three years after their introduction. RESULTS: Eighty per cent of the physicians answered and 92.5% were aware of the vaccination guidelines but only 2 out of 4 respected the targeted vaccination in young adults even when the vaccine was available. A policy of pertussis vaccination was applied only in 12 institutions, but even in these, the rate of vaccinated healthcare workers remained low or was not documented. CONCLUSION: Pertussis is a potential risk to newborns not or partially vaccinated in France. Even if the vaccine is available, adherence to pertussis vaccination guidelines must be improved. Efforts should be made to better publicize and apply pertussis vaccination guidelines. PMID- 19285818 TI - Concept of a selective tumour therapy and its evaluation by near-infrared fluorescence imaging and flat-panel volume computed tomography in mice. AB - Conventional chemotherapy of cancer has its limitations, especially in advanced and disseminated disease and suffers from lack of specificity. This results in a poor therapeutic index and considerable toxicity to normal organs. Therefore, many efforts are made to develop novel therapeutic tools against cancer with the aim of selectively targeting the drug to the tumour site. Drug delivery strategies fundamentally rely on the identification of good-quality biomarkers, allowing unequivocal discrimination between cancer and healthy tissue. At present, antibodies or antibody fragments have clearly proven their value as carrier molecules specific for a tumour-associated molecular marker. This present review draws attention to the use of near-infrared fluorescence (NIRF) imaging to investigate binding specificity and kinetics of carrier molecules such as monoclonal antibodies. In addition, flat-panel volume computed tomography (fpVCT) will be presented to monitor anatomical structures in tumour mouse models over time in a non-invasive manner. Each imaging device sheds light on a different aspect; functional imaging is applied to optimise the dose schedule and the concept of selective tumour therapies, whereas anatomical imaging assesses preclinically the efficacy of novel tumour therapies. Both imaging techniques in combination allow the visualisation of functional information obtained by NIRF imaging within an adequate anatomic framework. PMID- 19285819 TI - Quantification of synovial and erosive changes in rheumatoid arthritis with ultrasound--revisited. AB - Synovitis is a predictive factor of irreversible changes in the joints, tendons, and ligaments in patients with rheumatoid arthritis (RA). Therefore, the early demonstration of reversible, pre-erosive inflammatory features to diagnose RA, the monitoring of disease activity, and the response to therapy are of great importance. Technical developments in ultrasound now allow the quantification of synovitis and erosions, and enable the assessment and follow-up of disease activity. However, both the subjective and objective quantification techniques are associated with different problems. This review article highlights the advantages and disadvantages of sonographic quantification, and revisits the somewhat controversial positions apparent in the current literature. Familiarity with the imaging findings and the scoring systems used to characterize erosive changes are prerequisites for considerably improving the detection and monitoring of synovitis and erosions. The role of ultrasound in the diagnostic approach to RA, particularly in the quantification of synovial and erosive changes, will be explored and the current literature will be reviewed. PMID- 19285820 TI - Bone metastasis in patients with non-small cell lung cancer: the diagnostic role of F-18 FDG PET/CT. AB - PURPOSE: To evaluate the performance of F-18 FDG PET/CT in the detection of bone metastasis in non-small cell lung cancer (NSCLC) patients. MATERIALS AND METHODS: Three hundred and sixty-two consecutive NSCLC patients who underwent F-18 FDG PET/CT scanning were retrospectively analyzed. Each image of PET/CT, combined CT, and PET was performed at 10 separate areas and interpreted blindly and separately. The sensitivity, specificity and accuracy of F-18 FDG PET/CT, combined CT and F-18 FDG PET were calculated and the results were statistically analyzed. RESULTS: Bone metastasis was confirmed in 82 patients with 331 positive segments based on the image findings and clinical follow-up. On patient-based analysis, the sensitivity of F-18 FDG PET/CT (93.9%) was significantly higher than those of combined CT (74.4%) and F-18 FDG PET (84.1%), respectively (p<0.05). The overall specificity and accuracy of combined CT, F-18 FDG PET, and F-18 FDG PET/CT were 90.7%, 93.2%, 98.9% and 87.0%, 91.2%, and 97.8%, respectively (compared with PET/CT, p<0.05). On segment-based analysis, the sensitivity of the three modalities were 79.5%, 94.3%, and 98.8%, respectively (compared with PET/CT, p<0.05). The overall specificity and accuracy of the three modalities were 87.9%, 89.2%, 98.6% and 84.5%, 91.2%, 98.7%, respectively (compared with PET/CT, p<0.05). CONCLUSION: F-18 FDG PET/CT is superior to F-18 FDG PET or combined CT in detecting bone metastasis of NSCLC patients because of the complementation of CT and PET. It is worth noting that the added value of F 18 FDG PET/CT may beneficially impact the clinical management of NSCLC. PMID- 19285821 TI - Automatic assessment of the knee alignment angle on full-limb radiographs. AB - In this study a fully automatic assessment of the knee alignment angles in full limb radiographs was developed and compared to manual standard of reference measurements in a prospective manner. The data consisted of 28 knees which were gathered from total-leg radiographs of 15 patients (12 males and 3 females with a mean age of 29.4+/-6.9 years) consecutively. For statistical evaluation, a leave one-out cross-validation was performed. The pattern recognition and consequently the fully automatic assessment were successful in all patients. The automatically measured angles highly correlated with the standard of reference (r=0.989). The mean absolute difference was 0.578 degrees (95% CI: 0.399-0.757 degrees ). 82% of the angles differed less than 1 degrees from the standard of reference, 46% differed less than 0.5 degrees and 31% differed less than 0.2 degrees . The automatic method showed a high agreement between repeated measurements (+0.515 degrees to -0.429 degrees ). The automatic assessment of alignment angles in full limb radiographs were equal to the manual assessment. No measurement related user interaction was necessary to achieve results. PMID- 19285822 TI - A sensitive method for the determination of entecavir at picogram per milliliter level in human plasma by solid phase extraction and high-pH LC-MS/MS. AB - Entecavir is a guanine nucleoside analogue used in the treatment of hepatitis B virus (HBV) infection. In this paper, we describe an LC-MS/MS method that was developed and validated for the quantitation of entecavir in human EDTA plasma with both high sensitivity (lower limit of quantitation (LLOQ) of 5 pg/mL) and a wide concentration range (5000-fold) intended for low dose ascending clinical studies. High enrichment was achieved by taking advantage of the excellent loading capacity and reproducibility of Oasis HLB 96-well solid phase extraction plate, which allowed 1 mL of plasma samples to be processed in two equal sequential loading steps. Lobucavir, a structural analogue, was used as the internal standard. A filtration step following the reconstitution proved to be vital for the method robustness. The analyte and internal standard were separated on an Xterra MS C18 column with a gradient elution and high-pH mobile phases. Analytes were detected by positive ion electrospray tandem mass spectrometry. The high-pH mobile phase provided both excellent analyte on-column retention and peak shape, leading to the desired sensitivity. Validation results show good intra assay (12.3%CV) and inter-assay (3.1%CV) precisions, and good assay accuracy (+/ 7.6%Dev). Recovery was high (approximately 80%), however, the large volume of plasma used did result in a considerable matrix effect (approximately 0.45) which was well compensated by the analog internal standard. The method was applied to sample analysis of a Phase I clinical study. PMID- 19285823 TI - An unusual cause of spontaneous paralysis. PMID- 19285824 TI - Emergency department control of blood pressure in intracerebral hemorrhage. AB - BACKGROUND: Early treatment of elevated blood pressure (BP) in patients presenting with spontaneous intracerebral hemorrhage (ICH) may decrease hematoma enlargement and lead to better neurologic outcome. STUDY OBJECTIVE: To determine whether early BP control in patients with spontaneous ICH is both feasible and tolerated when initiated in the Emergency Department (ED). METHODS: A single center, prospective observational study in patients with spontaneous ICH was performed to evaluate a protocol to lower, and maintain for 24 h, the mean arterial pressure (MAP) to a range of 100-110 mm Hg within 120 min of arrival to the ED. An additional goal of placing a functional arterial line within 90 min was specified in our protocol. Hematoma volume, neurologic disability, adverse events, and in-hospital mortality were recorded. RESULTS: A total of 22 patients were enrolled over a 1-year study period. The average time to achieve our target MAP after implementation of our protocol was 123 min (range 19-297 min). The average time to arterial line placement was 84 min (range 36-160 min). Overall, 77% of the patients tolerated the 24-h protocol. The in-hospital mortality rate in this group of patients was 41%. CONCLUSIONS: Adopting a protocol to reduce and maintain the MAP to a target of 100-110 mm Hg within 120 min of ED arrival was safe and well tolerated in patients presenting with spontaneous ICH. If future trials demonstrate a clinical benefit of early BP control in spontaneous ICH, EDs should implement similar protocols. PMID- 19285825 TI - The impact of communication barriers on diagnostic confidence and ancillary testing in the emergency department. AB - BACKGROUND: Communication barriers (CBs) compromise the diagnostic power of the medical interview and may result in increased reliance on diagnostic tests or incorrect test ordering. The prevalence and degree to which these barriers affect diagnosis, testing, and treatment are unknown. STUDY OBJECTIVES: To quantify and characterize CBs encountered in the Emergency Department (ED), and assess the effect of CBs on initial diagnosis and perceived reliance on ancillary testing. METHODS: This was a prospective survey completed by emergency physicians after initial adult patient encounters. CB severity, diagnostic confidence, and reliance on ancillary testing were quantified on a 100-mm Visual Analog Scale (VAS) from least (0) to most (100). RESULTS: Data were collected on 417 ED patient encounters. CBs were reported in 46%; with a mean severity of 50 mm on a 100-mm VAS with endpoints of "perfect communication and "no communication." Language was the most commonly reported form of CB (28%). More than one CB was identified in 6%. The 100-mm VAS rating of diagnostic confidence was lower in patients with perceived CBs (64 mm) vs. those without CBs (80 mm), p < 0.001. VAS ratings of physician reliance on ancillary testing was higher in patients with perceived CBs (50 mm) vs. patients without a perceived CB (38 mm), p < 0.001. CONCLUSIONS: Communication barriers in our ED setting were common, and resulted in lower diagnostic confidence and increased perception that ancillary tests are needed to narrow the diagnosis. PMID- 19285827 TI - Sentinel lymph node biopsy in papillary thyroid cancer: comparison study of blue dye method and combined radioisotope and blue dye method in papillary thyroid cancer. AB - AIM: Occult lymph node metastasis is common in differentiated thyroid cancer (DTC). However, the role of lymph node dissection in the treatment of DTC remains controversial. The authors investigated the usefulness of methylene blue dye only method and combined radioisotope and methylene blue dye method for detecting SLN and compared the values of these two methods in patients with DTC. METHODS: From February to May 2008, 97 patients with DTC underwent sentinel lymph node biopsy (SLNB). The methylene blue dye method (dye only method) was used in 54 of the 97 patients, and radioisotope and methylene blue dye method (combined method) in 43 patients. RESULTS: The SLNs were identified in 89 patients, and the sensitivity and specificity of SLNB in the 97 patients were 85% and 100% respectively. For the dye only method, sensitivity, specificity, and the false negative rate (FNR) were 79%, 100%, and 21%; and for the combined method (43 patients) the corresponding figures were, 91%, 100%, and 9%, respectively. Six patients with SLN metastasis in the lateral neck underwent additional modified radical neck dissection (MRND). CONCLUSIONS: SLNB was found to be feasible, repeatable, and accurate in evaluating the lymph node status in patient with DTC. The present study indicates that the combined method could reduce false negative rate and increase detection rates of sentinel lymph node metastases, especially in lateral neck, compared to the dye only method. PMID- 19285828 TI - Quantitative intra-operative assessment of peritoneal carcinomatosis - a comparison of three prognostic tools. AB - AIMS: Selecting patients for cytoreductive surgery and hyperthermic intra peritoneal chemotherapy (HIPEC) remains challenging. We compared the predictive power of three intra-operative assessment tools of peritoneal involvement of colorectal cancer. METHODS: Ninety-two procedures (1999-2005) were prospectively scored using the Simplified Peritoneal Cancer Index (SPCI) and 7 Region Count. The Peritoneal Cancer Index (PCI) was retrospectively scored using the SPCI tool, operative notes and pathological reports. Endpoints were completeness of cytoreduction and overall survival. Logistic regression and Receiver Operating Characteristic (ROC) curves were applied to compare the predictive value of the three scoring systems on completeness of cytoreduction. RESULTS: After a median follow-up of 31 months, the median overall survival was 25.6 months. It decreased to 7.3 months, when cytoreduction was incomplete (p=0.001). An increased PCI, SPCI or number of regions were all associated with a decrease in probability of complete cytoreduction (p<0.05). With complete cytoreduction as outcome, the ROC areas for the PCI, SPCI and 7 Region Count were 0.92, 0.94 and 0.90, respectively (p=0.14). Using a cut-off value of 16 in the PCI system (p=0.03), 13 in the SPCI system (p=0.04) and 6 regions in the 7 Region Count (p=0.0002) the probability of complete cytoreduction decreased significantly. CONCLUSION: The PCI, SPCI and 7 Region Count are useful and equally effective prognostic tools predicting completeness of cytoreduction and associated improved survival. The 7 Region Count may be preferred due to its practical simplicity. PMID- 19285830 TI - A technology-assisted learning setup as assessment supplement for three persons with a diagnosis of post-coma vegetative state and pervasive motor impairment. AB - Post-coma persons in an apparent condition of vegetative state and pervasive motor impairment pose serious problems in terms of assessment and intervention options. A technology-based learning assessment procedure might serve for them as a diagnostic supplement with possible implications for rehabilitation intervention. The learning assessment procedure adopted in this study relied on hand-closure and eye-blinking responses and on microswitch technology to detect such responses and to present stimuli. Three participants were involved in the study. The technology consisted of a touch/pressure sensor fixed on the hand or an optic sensor mounted on an eyeglasses' frame, which were combined with a control system linked to stimulus sources. The study adopted an ABABCB sequence, in which A represented baseline periods, B intervention periods with stimuli contingent on the responses, and C a control condition with stimuli presented non contingently. Data showed that the level of responding during the B phases was significantly higher than the levels observed during the A phases as well as the C phase for two of the three participants (i.e., indicating clear signs of learning by them). Learning might be deemed to represent basic levels of knowledge/consciousness. Thus, detecting signs of learning might help one revise a previous diagnosis of vegetative state with wide implications for rehabilitation perspectives. PMID- 19285831 TI - Focal hand dystonia: a historical perspective from a clinician scholar. PMID- 19285832 TI - Focal hand dystonia: effectiveness of a home program of fitness and learning based sensorimotor and memory training. AB - STUDY DESIGN: This was a pre post test design. INTRODUCTION: Retraining the brain is one approach to remediate movement dysfunction resulting from task specific focal hand dystonia (FHD(TSP)). PURPOSE: Document change in task specific performance (TSP) for patients with FHD(TSP) after 8 weeks of comprehensive home training (fitness activities, task practice, learning based memory and sensorimotor training). METHODS: Thirteen subjects were admitted and evaluated at baseline, immediately and 6 months post treatment for task specific performance, functional independence, sensory discrimination, fine motor speed and strength. In Phase I, 10 subjects were randomly assigned to home training alone or supervised practice prior to initiating the home training. In phase II, 2 subjects crossed over and 3 new subjects were added (18 hands). The intent to treat model was followed. Outcomes were summarized by median, effect size, and proportion improving with nonparametric analysis for significance. RESULTS: Immediately post-intervention, TSP, sensory discrimination, and fine motor speed improved 60-80% (p<0.00l respectively). Functional independence and strength improved by 50%. Eleven subjects (16 hands) were re-evaluated at 6 months; all but one subject reported a return to work. Task-specific performance was scored 84-90%. Supervised practice was associated with greater compliance and greater gains in performance. CONCLUSIONS: Progressive task practice plus learning based memory and sensorimotor training can improve TSP in patients with FHD(TSP). Compliance with home training is enhanced when initiated with supervised practice. PMID- 19285833 TI - Effects of bamboo shoot consumption on lipid profiles and bowel function in healthy young women. AB - OBJECTIVE: This study evaluated the short-term effect of bamboo shoot consumption as a dietary fiber source on blood glucose, lipid profiles, hepatic function, and constipation symptoms in healthy women. METHODS: Eight subjects, 21- to 23-y-old women, with normal health status received a dietary fiber-free diet (control), a diet containing 25 g of cellulose, and a diet containing 360 g of bamboo shoots, with each diet segment lasting 6 d. At the end of each diet, blood biochemical parameters, such as glucose, triacylglycerols, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, and atherogenic index were measured and a questionnaire test for the evaluation of fecal excretion was taken. For statistical analysis, analysis of variance was performed. RESULTS: Serum total cholesterol, low-density lipoprotein cholesterol, and the atherogenic index were decreased with the bamboo shoot diet feeding compared with the dietary fiber-free diet. There were no differences in serum glucose levels among the tested diets. Fecal volume and bowel movement frequency in subjects fed the bamboo shoot diet were significantly increased. CONCLUSION: Bamboo shoots as a dietary fiber source has beneficial effects on lipid profile and bowel function. PMID- 19285834 TI - Relationship between hypometabolic patterns and ictal scalp EEG patterns in patients with unilateral hippocampal sclerosis: An FDG-PET study. AB - This study was to explore the relationship between scalp ictal EEG patterns and interictal hypometabolic patterns in hippocampal sclerosis-associated mesial temporal lobe epilepsy (HS-MTLE) and determine the clinical significance of interictal hypometabolic patterns. Twenty-five patients were classified into 2 groups based on initial ictal discharge (IID) frequency on scalp EEG: (a) those with a sustained regular 5- to 9-Hz rhythm with a restricted temporal or subtemporal distribution (group 1, N=9); and (b) those with an irregular 2- to 5 Hz rhythm with a widespread distribution (group 2, N=16). Using statistical parametric mapping, the PET results of each group were compared with age- and sex matched controls to identify regions of significant hypometabolism, and the clinical characteristics were compared. Group 1 showed focal hypometabolism confined to the ipsilateral temporal lobe, whereas group 2 showed widespread hypometabolism in the ipsilateral temporal lobe, insular cortex and anterior part of the putamen. The two groups showed no significant differences in clinical characteristics. Among semiologic features, dystonic limb posturing was more frequently observed in group 2 (p=0.03). In summary, scalp EEG IID patterns in HS MTLE can be important in determining hypometabolic patterns on interictal PET. Differences in hypometabolic patterns may reflect preferential pathways of ictal propagation rather than intrinsic epileptogenic regions. PMID- 19285835 TI - Antimicrobial resistance in non-tuberculous mycobacteria in New South Wales, 2002 2008. PMID- 19285836 TI - How can antipsychotics cause Diabetes Mellitus? Insights based on receptor binding profiles, humoral factors and transporter proteins. AB - The prevalence of Diabetes Mellitus (DM) is becoming a serious public health problem. The use of atypical antipsychotics has been associated with disruption of the glucose metabolism and therefore with causing DM. The underlying mechanisms are unknown, but knowledge of the differences between the pharmacological features of various antipsychotics combined with their diabetogenic profile might help us to understand those mechanisms. This article describes how the binding of various essential receptors or transporters in essential body tissues, adipose tissue, pancreatic tissue and liver and skeletal muscle tissue can cause disruption of the glucose metabolism. With such knowledge in mind one can try to explain the differences between the diabetogenic propensities of various antipsychotics. It is well known that clozapine and olanzapine cause weight gain and DM, whereas aripiprazole and ziprasidone have much less disruptive clinical profiles. The most significant risk factor for adiposity seems to be strong blocking of histaminergic receptors. An agonistic activity on serotonergic-1a receptors, with a very low affinity for muscarinergic 3 receptors, might protect against the development of DM. More data will become available which may help to solve the puzzle. PMID- 19285837 TI - Antiadhesive action of a marine microbial surfactant. AB - The antiadhesive action of a lipopeptide biosurfactant from a marine bacterium was investigated. The effect of cultivation conditions on the adhesion property of few bacterial strains was studied. It was observed that the static cultures showed greater adhesion due to scarcity of oxygen. The biosurfactant upon surface conditioning was found to be effective in removal of the microbial adhesion at a concentration as low as 0.1 g L(-1). The percentages of inhibition of adhesion against different test bacterial strains ranged from 15 to 89% using 0.1-10 g L( 1) of purified biosurfactant. These percentages of adhesion inhibition were found to be significantly higher than the previously reported values. The antiadhesive efficacy of the biosurfactant was also evident from confocal laser scanning microscopy studies. PMID- 19285838 TI - Synthesis of TiO2 nanoparticles using microorganisms. AB - A low-cost green and reproducible microbes (Lactobacillus sp. and Sachharomyces cerevisae) mediated biosynthesis of TiO(2) nanoparticles is reported. The synthesis is performed akin to room temperature in the laboratory ambience. X-ray and transmission electron microscopy analyses are performed to ascertain the formation of TiO(2) nanoparticles. Individual nanoparticles as well as a few aggregate having the size of 8-35 nm are found. Concentric Scherrer rings in the selected area electron diffraction pattern indicated that the nanoparticles are having all possible orientations. A possible involved mechanism for the biosynthesis of nano-TiO(2) has also been proposed in which pH as well as partial pressure of gaseous hydrogen (rH(2)) or redox potential of the culture solution seems to play an important role in the process. PMID- 19285839 TI - Public health implications of wireless technologies. AB - Global exposures to emerging wireless technologies from applications including mobile phones, cordless phones, DECT phones, WI-FI, WLAN, WiMAX, wireless internet, baby monitors, and others may present serious public health consequences. Evidence supporting a public health risk is documented in the BioInitiative Report. New, biologically based public exposure standards for chronic exposure to low-intensity exposures are warranted. Existing safety standards are obsolete because they are based solely on thermal effects from acute exposures. The rapidly expanding development of new wireless technologies and the long latency for the development of such serious diseases as brain cancers means that failure to take immediate action to reduce risks may result in an epidemic of potentially fatal diseases in the future. Regardless of whether or not the associations are causal, the strengths of the associations are sufficiently strong that in the opinion of the authors, taking action to reduce exposures is imperative, especially for the fetus and children. Such action is fully compatible with the precautionary principle, as enunciated by the Rio Declaration, the European Constitution Principle on Health (Section 3.1) and the European Union Treaties Article 174. PMID- 19285840 TI - Hypoxia-HIF-1alpha-C/EBPalpha/Runx1 signaling in leukemic cell differentiation. AB - Acute myeloid leukemia (AML), a class of prevalent hematopoietic malignancies, is caused by the acquisition of gene mutations that confer deregulated proliferation, impaired differentiation and a survival advantage of hematopoietic progenitors. More recently, we reported that cobalt chloride (CoCl(2))/iron chelator desferrioxamine (DFO)-mimicked hypoxia or moderate hypoxia (2% and 3% O(2)) can directly trigger differentiation of many subtypes of AML cells. Also, intermittent hypoxia significantly prolongs the survival of the transplanted leukemic mice with differentiation induction of leukemic cells. Additionally, these hypoxia-simulating agents selectively stimulate differentiation in acute promyelocytic leukemic cells induced by arsenic trioxide, an effective second line drug for this unique type of leukemia. Based on this interesting evidence in vitro and in vivo, the ongoing investigations showed the role of hypoxia inducible factor-1alpha (HIF-1alpha) protein through its non-transcriptional activity in myeloid cell differentiation, as evidenced by chemical interference, the conditional HIF-1alpha induction, the specific short hairpin RNAs (shRNAs) against HIF-1alpha and HIF-1beta, an essential partner for transcription activity of HIF-1. Furthermore, HIF-1alpha and two hematopoietic transcription factors CCAAT/enhancer binding protein alpha (C/EBPalpha) and Runx1/AML1 interact directly with each other. Such interactions increase the transcriptional activities of C/EBPalpha and Runx1/AML1, while C/EBPalpha competes with HIF-1beta for direct binding to HIF-1alpha protein, and significantly inhibits the DNA binding ability of HIF-1. As a protein is rapidly responsive to all-trans retinoic acid (ATRA), a classical clinical differentiation-inducing drug for AML, HIF-1alpha also plays a role in ATRA-induced differentiation of leukemic cells. PMID- 19285841 TI - Genotoxic effects of radiofrequency electromagnetic fields. AB - 101 publications are exploited which have studied genotoxicity of radiofrequency electromagnetic fields (RF-EMF) in vivo and in vitro. Of these 49 report a genotoxic effect and 42 do not. In addition, 8 studies failed to detect an influence on the genetic material, but showed that RF-EMF enhanced the genotoxic action of other chemical or physical agents. The controversial results may in part be explained by the different cellular systems. Moreover, inconsistencies may depend from the variety of analytical methods being used, which differ considerably with respect to sensitivity and specificity. Taking altogether there is ample evidence that RF-EMF can alter the genetic material of exposed cells in vivo and in vitro and in more than one way. This genotoxic action may be mediated by microthermal effects in cellular structures, formation of free radicals, or an interaction with DNA-repair mechanisms. PMID- 19285842 TI - Scorpion venom polypeptide accelerate irradiated hematopoietic cells proliferation. AB - OBJECTIVE: To study the effects of Scorpion venom polypeptide (SVP) on the irradiated hematopoietic progenitor cells and the initial research of its mechanism. METHODS AND MATERIALS: (1) MTT array was used to select the effective concentration of SVP that had proliferate action on the irradiated early hematopoietic cells (K562), just like the doses of experiment in vitro; (2) The male BALB/c mice were divided into NS control group, SVP IV group and SVP V group. After treatment and sublethal irradiation, the C-KIT and IL-6Ralpha levels of bone marrow cells were detected by immunohistochemistry and tissue array; (3) The bone marrow cells of the normal BALB/c mice, given to SVP IV and SVP V after different action times respectively, were taken to extract the total proteins inside the cell, the phosphorylated STAT3 protein levels in JAK-STAT signal transduction pathway were detected by Western blot array. RESULTS: (1) 30mg/L SVP IV has an obvious effect to accelerate K562 cell proliferation; (2) The C-KIT and IL-6Ralpha expression on bone marrow cell surfaces in SVP IV and SVP V groups were negative (control with the saline group, p>0.05); (3) The phosphorylated STAT3 protein levels in bone marrow cells of SVP IV group had a rise-and-fall trend within 30min, while the test of SVP V group showed that the phosphorylated STAT3 protein levels obviously elevated after 30min. CONCLUSIONS: The results show that certain SVP IV concentration can protect the hematopoietic progenitor cells after irradiation, and the underlying mechanism of SVP accelerating the hematopoietic recovery in irradiated mice may be related to the activation of the JAK-STAT signal pathway. PMID- 19285843 TI - Hypermethylation of estrogen receptor-alpha gene in atheromatosis patients and its correlation with homocysteine. AB - OBJECTIVE: To investigate the aberrant DNA methylation in promoter region of estrogen receptor alpha (ERalpha) in atherosclerosis (As) and the possible involvement of homocysteine (Hcy) in its pathogenesis. METHODS: The blood samples were collected from 54 patients with As approved by carotid colorized ultrasound examination and 28 healthy control subjects. The methylation status of CpG islands in ER-alpha gene promoter region of genome DNA was analyzed by nested methylation-specific PCR (nMSP). tHcy was examined by fluorescent-biochemical method. Spearman rank correlation was used to analyse the relationship between the degree of methylation in ER-alpha gene and the level of tHcy. Cultured smooth muscle cells of Homo sapiens were treated by Hcy with different concentrations and different treating time, again the DNA methylation status was assayed by nMSP, and the proliferation of SMC was assayed by MTT. RESULTS: Hypermethylation of ER-alpha gene promoter region was found in 38 cases of atherosclerosis patients, and the methylation frequency was 70.4%. While in healthy controls, just 8 of 28 samples hypermethylation was found, only 28.6% methylation frequency was detected, much lower than the one in atherosclerosis group (p<0.05). Meanwhile, the level of tHcy in atherosclerosis group was significantly higher than that in control group (p<0.05). The spearman rank correlation analysis explored an obvious correlation between the degree of methylation in ER-alpha gene and the level of tHcy (r=0.809, p<0.05), and the severity of atherosclerotic lesion was also heightened along with the increment of plasma level of tHcy. The cultured SMCs treated by Hcy resulted in de novo methylation in promoter region of ERalpha gene with a concentration and treating time-dependent manner, and a dose-dependent promoting effect on SMC proliferation. The in vivo and in vitro data coincidently showed that the Hcy could promote the hypermethylation of ERalpha gene, which may be an important mechanism for the pathogenesis of As. CONCLUSION: Hypermethylation of CpG islands in ER-alpha gene promoter region was found in much higher frequency in atherosclerosis patients, which is positively correlated with the increased level of plasma tHcy and the severity of atherosclerotic lesion, and the in vitro experimental results further extended above clinical data that HHcy can lead to the hypermethylation of ER-alpha gene, and hence to promote the occurrence and development of As. PMID- 19285844 TI - Effect of vitamin C in reducing the toxicity of endosulfan in liver in rabbits. AB - In this study, the effect of endosulfan, an organochlorine pesticide, and the ameliorating effect of vitamin C on the livers of New Zealand white rabbits were studied. Livers of the rabbits were examined grossly and histopathologically, and caspase-3 activity was detected by immunohistochemical methods. A total of twenty four rabbits were divided into four groups (n=6). Rabbits in Group I (END) were daily given a sublethal dose of endosulfan (1mg/kgbw) in corn oil by oral gavage for 6 weeks. Group II (END+C) received the same dose of endosulfan and additionally Vit C (20mg/kgbw) every other day during this period. Group III (OIL+C) received corn oil daily by oral gavage and vitamin C every other day for 6 weeks. Group IV (OIL), the control group, received only corn oil daily, by oral gavage throughout the experiment. The concentration of alpha-endosulfan in the END group was higher in livers (0.102+/-0.012ppb) than the beta-endosulfan (0.072+/-0.001ppb). Decreased accumulation of alpha and beta endosulfan was observed in the END+C group (0.025+/-0.003 and 0.016+/-0.002ppb, respectively) (p<0.0001). The most prominent gross findings at the necropsy were seen in the END group, in which swollen and pale livers were commonly observed. Hemorrhages, degenerations, necrosis, and in some rabbits bile duct hyperplasia were the marked histopathological findings of the END group. Caspase-3 positive reaction was more severe in this group than in the others. An ameliorating effect of Vit C on gross, histopathological and immunohistochemical findings was observed in the END+C group. The results revealed that endosulfan is highly toxic for rabbit livers. However, toxicity was decreased by Vit C treatment, which reduced the accumulation of endosulfan in livers four-fold. PMID- 19285845 TI - 1st international ESTP expert workshop: "Larynx squamous metaplasia". A re consideration of morphology and diagnostic approaches in rodent studies and its relevance for human risk assessment. AB - Invited international experts participated in a 2-day workshop organized by the European Society of Toxicologic Pathology (ESTP) to evaluate and discuss spontaneous and induced laryngeal lesions in rodents. The main purpose of the workshop was to agree upon the terminology and relevance of a range of laryngeal changes that varied from very subtle epithelial alterations up to severe metaplastic or neoplastic lesions. The workshop experts concluded that minimal, focal epithelial changes of the laryngeal epithelium, predominantly occurring at the base of the epiglottis, should be given the descriptive term of "epithelial alteration" and assessed as "non-adverse". Although observed as induced effects they may also occur in non-treated animals and were not considered to have a potential for a laryngeal dysfunction. Also, cases of minimal to slight laryngeal squamous metaplasia that are not observed diffusely could occur spontaneously or as treatment-induced lesions and should be assessed as "non-adverse". Cases of moderate to severe laryngeal squamous metaplasia observed diffusely in multiple levels should be regarded as "adverse", as there is a potential for dysfunction of the larynx. The occurrence of dysplasia or cellular atypia linked to laryngeal squamous metaplasia should always be reported separately and described in detail. In the evaluation of treatment-related effects of the larynx in studies utilizing aged animals, it has to be considered that moderate or even severe cases of focal laryngeal squamous metaplasia may occasionally be found as age-related, spontaneous lesions. Although inhalation exposure of rodents to non-genotoxic compounds may cause laryngeal squamous metaplasia, none of the workshop experts were aware of any reported cases of tumor induction in the larynx with a non genotoxic compound. Therefore, for non-genotoxic compounds, the workshop experts did not regard laryngeal squamous metaplasia by itself as a precancerous lesion. PMID- 19285846 TI - Cardioprotective effect of 'Khamira Abresham Hakim Arshad Wala' a unani formulation in isoproterenol-induced myocardial necrosis in rats. AB - The present study was designed to investigate whether Khamira Abresham Hakim Arshad Wala (KAHAW), a preparation of Unani System of Medicine, is able to attenuate the isoproterenol (ISO)-induced myocardial necrosis on the basis of its effects on hemodynamic, antioxidant, histopathological and ultrastructural parameters. Male Wistar albino rats were administered KAHAW (200, 400 and 800mg/kg/day, orally) or vehicle for 14 days with concurrent ISO administration (85mg/kg, subcutaneously, 2 doses at 24h interval) on 13th and 14th day. On the 15th day, vehicle+ISO-treated rats exhibit cardiac dysfunctions as indicated by decrease in systolic, diastolic, and mean arterial pressures, reduction in both maximum positive and maximum negative rates of developed left ventricular pressure (+/-LVdp/dt) and an increase in left ventricular end-diastolic pressure (LVEDP). Biochemical analysis of their heart homogenate presented reduced levels of enzymes viz., superoxide dismutase (SOD), catalase (CAT), lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB) isoenzyme. A marked reduction in reduced glutathione (GSH) levels along with increase in levels of thiobarbituric acid reactive substances (TBARS) was also observed in rat myocardium. Myocardial necrosis, edema and inflammation were evident from the light microscopic and ultrastructural changes. KAHAW at dose of 800mg/kg/day significantly reversed majority of hemodynamic and antioxidant derangements. The protective role of KAHAW on ISO-induced myocardial necrosis was further confirmed by histopathological and ultrastructural examination. There was no significant change in heart rate in all experimental groups. KAHAW per se groups showed no significant change when compared with vehicle control group. The study results thus demonstrated the cardioprotective potential of KAHAW against ISO-induced myocardial necrosis and associated oxidative stress. PMID- 19285847 TI - Walther Flemming on histology in medicine 1878: a newly discovered letter to his father. AB - Histology is a child of the 19th century, and its status in medicine was long in doubt. Was "microscopic anatomy" to be viewed as a refinement of the traditional discipline or as a technically complex subject of doubtful practical value? This was one of the questions that faced the commission charged with reforming examinations in German medical schools in 1878. One of its members, Carl Friedrich Flemming [1799-1880], was able to refer this matter to an acknowledged expert, his son Walther Flemming, who was Professor of Anatomy and Histology at the University of Kiel. Walther's views are contained in a letter that was recently discovered among Carl Friedrich's papers held in the State Library of Mecklenburg-Vorpommern in Schwerin. To the best of our knowledge, this letter is the only surviving piece of personal correspondence from the hand of Walther Flemming, one of the most distinguished biologists of his time. The original text, together with an annotated English translation, is published here for the first time. The letter throws new light on Flemming's attitudes to medical education, and also illuminates his relationship with his father, himself a figure of considerable stature in psychiatric medicine in 19th-century Germany. The younger Flemming [1843-1905], an acknowledged master of microscopy, made his most distinguished contributions to the field we now call cell biology. In addition to the terms chromatin and mitosis, we owe to him the first comprehensive account of the latter, as well as the fundamental insight that meiosis involves two successive divisions. PMID- 19285848 TI - Adoptive immunotherapy of cancer using CD4(+) T cells. AB - CD4(+) T cells are central to the function of the immune system but their role in tumor immunity remains underappreciated. It is becoming clear that there is an enormous diversity of CD4(+) T cell polarization patterns including Th1, Th2, Th17, and regulatory T cells (Tregs). These functionally divergent T cell subsets can have opposing effects -- they can trigger tumor rejection or inhibit treatment after adoptive cell transfer. Some polarized CD4(+) cells have plasticity, and their phenotypes and functions can evolve in vivo. Recent advances in understanding of polarization and differentiation of lymphocytes, as well as some intriguing developments in the clinic, indicate that the use of CD4(+) T cell subsets in the immunotherapy of cancer has unrealized potential. PMID- 19285849 TI - Effect of mouth taping at night on asthma control--a randomised single-blind crossover study. AB - BACKGROUND: Nose breathing ensures that inspired air is warm, filtered and moist and may therefore benefit patients with asthma. It features in some complementary approaches to treat asthma and is encouraged at night in the Buteyko technique by the use of mouth taping. In this pragmatic study we sought to determine whether taping the mouth at night has any effect on asthma control compared with usual breathing in patients with symptomatic asthma, since if it was effective it would be a simple intervention to implement. METHODS: This was a randomised, single blind, crossover study of participants (n=51) with symptomatic asthma (mean FEV(1) 86% predicted). A 4-week period of usual breathing at night was followed by use of mouth taping with microporous tape, as in the Buteyko technique, or vice versa, with a 2-week run-in period and a minimum 2-week washout period of usual breathing between 'treatments'. Primary outcomes were morning peak expiratory flow and symptom scores (Asthma Control Diary). Outcomes were measured and analysed without knowledge of treatment allocation. RESULTS: Fifty participants completed the study and reported taping their mouth for a median 26 of 28 nights. Although 36 participants said mouth taping was very or fairly acceptable there were no differences between treatments for morning peak expiratory flow (mean difference -1l/min (95%CI, -9 to 7)) or symptoms scores (mean difference -0.12 (95%CI, -0.30 to 0.06)) nor for any secondary measures. CONCLUSIONS: Taping the mouth at night had no effect on asthma control in patients with symptomatic asthma. PMID- 19285850 TI - Smoking cessation in chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is increasing in prevalence, and is predicted to become the third leading cause of deaths worldwide by 2020. The precise prevalence of COPD is not known, as many individuals with the disease are left undiagnosed, despite the requirement of only simple spirometry testing for disease detection. The major risk factor for the development of COPD is cigarette smoking, with 90% of deaths from COPD directly attributable to smoking. Therefore smoking cessation is the most effective means of halting or slowing the progress of this disease. This review summarizes and compares the differential characteristics of smokers with COPD vs. those without COPD in relation to their smoking behavior and quitting attempts, and discusses the various strategies that can be used to help patients quit and improve their likelihood of long-term smoking cessation. Of the various behavioral interventions available that can increase the likelihood of smoking cessation, one of the simplest and most effective strategies that physicians can use is simply to advise their patients to quit, particularly if this advice is combined with informing the patients of their "lung age". We also discuss the pharmacologic therapies used to enhance the likelihood of quitting, including nicotine replacement, bupropion SR and varenicline, along with novel nicotine vaccines, which are currently undergoing clinical trials. PMID- 19285851 TI - Translational control from head to tail. AB - mRNA translation, a highly coordinated affair involving many proteins and RNAs, is generally divided into three steps: initiation, elongation, and termination. Each of these steps serves as a point of regulation to control the amount of protein that is produced. The protein 4E-HP has recently been shown to disrupt recruitment of the translation initiation complex by directly binding the 5' cap of cellular mRNAs. Recent work has shown elongation rates are probably altered during mitosis and certain types of synaptic transmission. Other work has shown premature termination of mRNAs lacking stop codons appears to repress their translation. Together, these studies highlight the importance of translational control in diverse processes such as development, cancer, and synaptic plasticity. PMID- 19285852 TI - Electrochemiluminescence from hydrophilic thin film Ru(bpy)3(2+)-modified electrode prepared using natural halloysite nanotubes and polyacrylamide gel. AB - Because Ru(bpy)(3)(2+)-modified electrodes are often used in aqueous condition, the development of a hydrophilic modified electrode is of critical importance. Herein a hydrophilic, thin film Ru(bpy)(3)(2+)-modified electrode is successfully developed using polyacrylamide gel to embed halloysite nanotubes on the electrode surface which is used to adsorb Ru(bpy)(3)(2+) by cation-exchange. X-ray photoelectron spectroscopy demonstrates the formation of the film on an electrode and the high adsorbing capacity of the halloysite nanotubes toward Ru(bpy)(3)(2+). The different electrochemiluminescence (ECL) behaviors of the electrode using nitrilotriacetic acid and tripropylamine, as co-reactants, illustrate the hydrophilic character of the modified electrode. Contrary to the previous works, the addition of carbon nanotubes into the modified electrode film leads to a decreased ECL emission, due to the reduction in the porosity of the film, which hinders the diffusion of the analyte. PMID- 19285853 TI - Novel U-bent fiber optic probe for localized surface plasmon resonance based biosensor. AB - The aim of this study is to develop an optical absorbance based biosensor suitable for wide scale use in resource-poor locales. A sensor for sensitive measurement of refractive index (RI) with the help of optical absorbance properties of gold nanoparticles (GNP) coupled to an efficient optical transducer in the form of a U-bent fiber optic probe is described. A U-bent probe was fabricated by a simple procedure. The absorbance due to the localized surface plasmon resonance (LSPR) of fiber-bound GNP was found to be linear to refractive index changes between 1.33 and 1.35. A U-bent probe of 200 microm diameter with a bend radius of 0.75 mm gave rise to a sensitivity of 35 DeltaA/RIU at 540 nm. The resolution of the sensor probe was 3.8x10(-5) RIU. Label-free biosensing was demonstrated using these probes with the help of IgG-anti IgG as bioreceptor analyte pair. PMID- 19285854 TI - A microelectrode-based sensor for label-free in vitro detection of ischemic effects on cardiomyocytes. AB - Heart diseases represent the most common cause of death in industrialised countries. For this reason target identification and development of novel anti target drugs are in the focus of pharmaceutical industry. Especially cardiac infarct is a topical field of research. A bottleneck in today's long-duration and high-cost drug development is the lack of fast, label-free and cell-based high throughput/high content screening (HTS/HCS) assays for bridging the gap between cell-free screening and animal experiments. Here, we report for the first time on an in vitro cardiac ischemic model, where pathological consequences of simulated cardiac infarct can be detected quantitatively by microelectrode array-based impedance spectroscopy. Using the contractile HL-1 cell line and defined ischemic conditions we were able to develop a standardised and reproducible pathologic model. We characterised and verified the HL-1 based ischemic model by apoptosis and proliferation assays as well as immunochemical analysis of cell-cell junctions. We showed that the observed cell and biomolecular effects correspond with results obtained by impedance spectroscopy. Functionality of the impedimetric assay was demonstrated by real-time detection of reduced pathological effects due to application of the selective Rac1 inhibitor NCS23766. Numerical analysis by means of an equivalent circuit allowed the quantification of changes in resistance and capacitance of the adherent cell layer after ischemic treatment and application of NSC23766 as drug model. Our findings provide a novel cell-based real-time screening system for testing drug candidates against cardiac infarct and its implications. PMID- 19285856 TI - Web-based resources for sun protection information--a French-language evaluation. AB - BACKGROUND: With the dramatic increase in ultraviolet-induced skin damage, advocating the benefits of reasonable and moderate exposure to sunlight is a public health priority in most Western countries. OBJECTIVE: The aim of this study was to investigate the nature and quality of the information posted on the Internet about the effects of the sun on health and about sun protection. METHODS: We used eight terms with eight search engines to identify relevant websites providing sun protection advice on the French-speaking web. This information was compared to the recommendations issued by nine official organisations. Influence of website quality and commercial links on the quality of the advice provided were evaluated. For the purposes of this comparison, we attributed marks to the following four criteria: 1: risks and benefits of sun exposure; 2: sun avoidance; 3: clothing recommendations; and 4: sunscreen use. RESULTS: The search identified 1551 links. After exclusion of sites duplicated either within or between search engines, and of websites used as references, we found 129 web pages dedicated to sun protection advice. Data on the effects of the sun and sun protection were deficient in most websites in comparison to official organisation websites. The results were superior if the web pages or websites respected quality criteria. Commercial links tended to score poor marks for 1, 2 and 3 but better marks for 4 (sunscreen use). CONCLUSION: We showed the limits of web-based resources on the French-speaking web regarding sun protection advice. The poor quality of most of the websites and commercial conflict of interest with sunscreen manufacturers may explain these limits. PMID- 19285855 TI - Novel genetic tools for studying food-borne Salmonella. AB - Nontyphoidal Salmonellae are highly prevalent food-borne pathogens. High throughput sequencing of Salmonella genomes is expanding our knowledge of the evolution of serovars and epidemic isolates. Genome sequences have also allowed the creation of complete microarrays. Microarrays have improved the throughput of in vivo expression technology (IVET) used to uncover promoters active during infection. In another method, signature tagged mutagenesis (STM), pools of mutants are subjected to selection. Changes in the population are monitored on a microarray, revealing genes under selection. Complete genome sequences permit the construction of pools of targeted in-frame deletions that have improved STM by minimizing the number of clones and the polarity of each mutant. Together, genome sequences and the continuing development of new tools for functional genomics will drive a revolution in the understanding of Salmonellae in many different niches that are critical for food safety. PMID- 19285857 TI - Estimation of an optimal chemotherapy utilisation rate for head and neck carcinoma: setting an evidence-based benchmark for the best-quality cancer care. AB - BACKGROUND: We estimated the optimal chemotherapy utilisation rate for head and neck cancer as a benchmark for measuring and improving the quality of cancer care. METHODS: An optimal chemotherapy utilisation tree was constructed using indications for chemotherapy that were identified from evidence-based treatment guidelines. Data on the proportion of patient and tumour-related attributes for which chemotherapy was indicated were obtained and merged with the treatment indications to calculate the optimal utilisation rate. The robustness of the model was tested with sensitivity analysis and Monte Carlo simulation. The optimal chemotherapy utilisation rate was compared with actual utilisation rates reported. RESULTS: Chemotherapy is indicated at least once in 36% (95% CI, 33 38%) of all patients with head and neck carcinoma. The optimal utilisation rates by subsites were as follows: lip, 8%; oral cavity, 40%; nasopharynx, 69%; oropharynx, 66%; hypopharynx, 74%; larynx, 43%; salivary gland, 48% and paranasal sinus with nasal cavity, 38%. CONCLUSIONS: The optimal proportion of patients who should receive chemotherapy in the head and neck carcinoma population has risen significantly over the past 20 years. This temporal rise does not appear to be reflected in the limited actual utilisation rates that are available for comparison. Large population-based studies are recommended to further assess the current practice and compliance to guideline recommended care. PMID- 19285858 TI - Late morbidity associated with a tumour-negative sentinel lymph node biopsy in primary breast cancer patients: a systematic review. AB - AIMS: To evaluate the relationship between late morbidity (i.e. > or =6 months) and a tumour-negative sentinel lymph node biopsy (SLNB) in primary breast cancer patients by using a systematic review approach, and to identify the predictors of late morbidity. METHODS: We performed a systematic review of the literature for studies concerning the late morbidity of patients who had undergone SLNB alone or SLNB followed by ALND when SLN metastases were found. A literature search over the last 16 years (1993-2008) was performed in the databases MEDLINE and EMBASE. The methodological quality of the selected studies was assessed according to a list of predefined criteria. The data of assessment and predictors of late morbidity were collected. RESULTS: We identified a total of 32 papers reporting 27 independent cohort studies, of which 17 were high quality studies and were further analysed in this review. There was a great variation in the prevalence of pain (7.5-36%), impairment of range of motion (0.0-31.0%), oedema (0.0-14.0%), decreased strength (11.0-19.0%) and sensory disorders (1.0-66.0%). Factors such as time after surgery and young age were strong predictors of late morbidity. Breast surgery, radiation to axilla, tumour location, body mass index (BMI) and two-step procedure, especially lymph mapping techniques, could also predict the late morbidity to different extents. CONCLUSIONS: SLNB-associated late morbidity, even with a low prevalence, remains a clinical problem which cannot be neglected in primary breast cancer patients. Time after surgery and young age are the important predictors for late morbidity in primary breast cancer patients after SLNB; breast surgery, radiation to axilla, tumour location, BMI and two-step procedure also have limited prognostic value. PMID- 19285859 TI - Immobilization of bioluminescent Escherichia coli cells using natural and artificial fibers treated with polyethyleneimine. AB - Biosensors based on whole-cell bioluminescence have the potential to become a cost-effective alternative to conventional detection methods upon validation of target selectivity and sensitivity. However, quantitative analysis of bioluminescence is greatly hindered due to lack of control over the total number of cells in a suspending culture. In this study, the effect of surface properties of genetically engineered luminous E. coli cells and fibrous matrices on the immobilization capacity and effectiveness under various environmental conditions were characterized. Four different fibers, including cotton, polyester, viscose rayon, and silk, were investigated. Although cell adhesion was observed on untreated viscose and cotton fibers, viscose fiber pretreated with 0.667% polyethyleneimine (PEI) was found capable of immobilizing the most viable E. coli DPD2234 cells, followed by viscose treated with 0.33% and 1% PEI. The cells immobilized on PEI-treated viscose remained viable and yielded 20% or more bioluminescence signals immediately upon contact with the inducer up to 72 h without feeding nutrients to the cells, suggesting that viscose treated with 0.667% PEI could provide a stable immobilization mechanism for bioluminescent E. coli cells with long sensing period, quick response time, and good signal reproducibility. PMID- 19285860 TI - Turn-on fluorescent probe with visible light excitation for labeling of hexahistidine tagged protein. AB - We report here the development of a novel fluorescein-based probe which shows selective fluorescence enhancement on binding to a hexahistidine-tagged protein. No fluorescence change was observed with untagged protein. This probe is excitable with visible light and is considered to be suitable for use in biological applications. PMID- 19285861 TI - Regioisomerism-dependent TLR7 agonism and antagonism in an imidazoquinoline. AB - Chronic immune activation is a hallmark of progressive HIV infection. Recent reports point to the engagement of toll-like receptor 7 (TLR7) and -9 by viral RNA as contributing to the activation of innate immune responses, which drive viral replication leading to immune exhaustion. The only known class of TLR7 antagonists is single-stranded phosphorothioate oligonucleotides, which has been demonstrated to inhibit immune activation in human and Rhesus macaque in vitro models. The availability of a selective and potent small-molecule TLR7 antagonist should allow the evaluation of potential benefits of suppression of TLR7-mediated immune activation in HIV/AIDS. Gardiquimod is a known N(1)-substituted 1H imidazoquinoline TLR7 agonist, the synthesis of which has not been published. We show that the 3H regioisomer is completely inactive as a TLR7 agonist and is weakly antagonistic. A des-amino precursor of the 3H regioisomer is more potent as a TLR7 antagonist, with an IC(50) value of 7.5 microM. This class of compound may serve as a starting point for the development of small-molecule inhibitors of TLR7. PMID- 19285862 TI - Piperidinyl-2-phenethylamino inhibitors of DPP-IV for the treatment of type 2 diabetes. AB - A highly ligand efficient lead molecule was rapidly developed into a DPP-IV selective candidate series using focused small library synthesis. A significant hurdle for series advancement was genetic safety since some agents in this series impaired chromosome division that was detected using the in vitro micronucleus assay. A recently developed high-throughput imaging-based in vitro micronucleus assay enabled the identification of chemical space with a low probability of micronucleus activity. Advanced profiling of a subset within this space identified a compound with a clean safety profile, an acceptable human DPP-IV inhibition profile based on the rat PK/PD model and a projected human dose that was suitable for clinical development. PMID- 19285863 TI - Structure-activity relationships of diverse annonaceous acetogenins against human tumor cells. AB - Twelve annonaceous acetogenins (ACGs) with different stereochemical structures and configuration were selected to test for their inhibitions on the growth of Hela, SMMC-7541, SGC-7901, MCF-7 and A-5408 tumor cell lines using MTT method. This was the first to simultaneously investigate effects of structural factors of stereochemical structures and configuration on cytotoxicities with structure activity relationship. The present study showed that cytotoxic selectivities of ACGs with threo/trans/threo/trans/erythro stereochemical arrangement were gently more active than those with threo/trans/threo/trans/threo stereochemical arrangement, and ACGs with cis THF ring partly produced notable cytotoxic selectivities. Furthermore, ACGs with S configuration at C-24 exhibited gently more cytotoxic selectivities potency than those with R configuration at C-24. PMID- 19285864 TI - Eosinophilic myositis in calpainopathy: could immunosuppression of the eosinophilic myositis alter the early natural course of the dystrophic disease? AB - An 11-year-old girl with a calpain-3 gene (CAPN-3) mutation and eosinophilic myositis on muscle biopsy had high serum CK levels and eosinophil counts which showed spontaneous fluctuations. After commencement of immunosuppressive therapy reciprocal changes occurred in response to alterations in doses of the medications. Subacutely evolving and spreading muscle weakness developed during tapering of the immunosuppressive medications. These observations suggest that either the occurrence of eosinophilic myositis or the withdrawal of the immunosuppressive treatment may have accelerated the clinical course of the calpainopathy in this case. The positive effect of immunosuppressive therapy might have implications for the management of calpainopathy with an inflammatory component. PMID- 19285865 TI - The m.3244G>A mutation in mtDNA is another cause of progressive external ophthalmoplegia. AB - We sequenced all mitochondrial tRNA genes in a 61-year-old man with chronic progressive external ophthalmoplegia and mitochondrial myopathy but without mtDNA rearrangements, and identified a heteroplasmic m.3244G>A mutation in the tRNA(Leu(UUR)) gene. This mutation had been previously associated with the MELAS phenotype, but not described in any detail. The mutation load in muscle was 84% and COX-negative fibers harbored greater levels of mutant genomes than COX positive fibers. The m.3244G>A mutation affects a highly conserved nucleotide in the dihydrouridine loop and has been associated with a wobble modification deficiency of the mutant tRNA. PMID- 19285866 TI - PRDM16: the interconvertible adipo-myocyte switch. AB - Both brown and white adipocytes were previously considered to be derived from the same precursor cell, despite being histologically and functionally different. However, a recent study shows that overexpression of the transcriptional regulator positive regulatory domain containing 16 (PRDM16) determines the development of brown adipocytes from a progenitor that expresses myoblast markers. Surprisingly, loss of PRDM16 from these precursors does not lead to white adipocyte differentiation. Thus, PRDM16 controls a bidirectional cell fate switch between skeletal myoblasts and brown adipocytes. PMID- 19285867 TI - Division plane control in plants: new players in the band. AB - Unique mechanisms are used to orient cell division planes in plants. A cortical ring of cytoskeletal filaments called the preprophase band (PPB) predicts the future division plane during G2 and is disassembled as the mitotic spindle forms, leaving behind a 'cortical division site' (CDS) that guides the placement of the new cell wall (cell plate) during cytokinesis. The molecular features of the CDS have remained elusive for decades. Recently, a few proteins have at last been identified that are specifically localized to or excluded from the CDS and that participate in the orientation, attachment or maturation of cell plates. Recent progress has also been made in identifying proteins needed for PPB formation and thus for division plane establishment. PMID- 19285868 TI - Concurrent cues for cytokinetic furrow induction in animal cells. AB - Animal cells are deformable, yet live together bound into tissues. Consequently, physical perturbations imposed by neighbors threaten to disrupt the spatial coordination of cell cleavage with chromosome segregation during mitosis. Emerging evidence demonstrates that animal cells integrate multiple positional cues during cleavage-furrow induction, perhaps to facilitate error correction. Classical work indicated that the asters provide the stimulus for furrow induction, but recent results implicate the central spindle at least as much. Similarly, although classical work concluded that the stimulus occurs at the cell equator, new evidence shows that asters modulate cortical contractility outside the equator as well. Meanwhile, a newly revealed distinction between stable and dynamic astral microtubules suggests that these subsets might have complementary effects on furrow induction. PMID- 19285869 TI - The actin cytoskeleton in spindle assembly and positioning. AB - The most dramatic changes in eukaryotic cytoskeletal organization and dynamics occur during passage through mitosis. Although both spindle self-organization and actin-dependent cytokinesis have long been the subject of intense investigation, it has only recently become apparent that the actin cortex also has a key role during early mitosis. This is most striking in animal cells, in which changes in the actin cytoskeleton drive mitotic cell rounding and cortical stiffening. This mitotic cortex then functions as a foundation for spindle assembly and to guide spindle orientation with respect to extracellular chemical and mechanical cues. Here, we discuss this recent work and the possible role of crosstalk between the mitotic actin cortex and the plus ends of astral microtubules in this process. PMID- 19285870 TI - Understanding the dopaminergic deficits in Parkinson's disease: insights into disease heterogeneity. AB - Parkinson's disease is a common condition with a broad clinical diversity suggesting the existence of distinct subgroups of patients. This paper describes how dopaminergic disruption within basal ganglia circuitry accounts for some of the major features of the disease and examines how the limited repertoire of the output nuclei within these pathways could allow for an element of "cross-talk" between competing inputs. It is proposed that such conditions could lead to an excessive inhibition of the thalamus and pedunculopontine nucleus and account for many of the familiar patterns of clinical phenotype. It is further postulated that this phenomenon may be acting via increased synchronization within the basal ganglia circuitry. PMID- 19285871 TI - Isolated cortical vein thrombosis in a patient with arteriovenous malformation. PMID- 19285872 TI - IKKbeta inhibitors identification part I: homology model assisted structure based virtual screening. AB - Control of NF-kappaB release through the inhibition of IKKbeta has been identified as a potential target for the treatment of inflammatory and autoimmune diseases. We have employed structure based virtual screening scheme to identify lead like molecule from ChemDiv database. Homology models of IKKbeta enzyme were developed based on the crystal structures of four kinases. The efficiency of the homology model has been validated at different levels. Docking of known inhibitors library revealed the possible binding mode of inhibitors. Besides, the docking sequence analyses results indicate the responsibility of Glu172 in selectivity. Structure based virtual screening of ChemDiv database has yielded 277 hits. Top scoring 75 compounds were selected and purchased for the IKKbeta enzyme inhibition test. From the combined approach of virtual screening followed by biological screening, we have identified six novel compounds that can work against IKKbeta, in which 1 compound had highest inhibition rate 82.09% at 10 microM and IC(50) 1.76 microM and 5 compounds had 25.35-48.80% inhibition. PMID- 19285873 TI - Application of gamma irradiation for the enhanced physiological properties of polysaccharides from seaweeds. AB - Polysaccharides from seaweeds, fucoidan and laminarin, were irradiated with gamma rays, and their structural changes and anti-oxidative activities were investigated. The gamma irradiation decreased the average molecular weights of polysaccharides, and UV spectra of irradiated polysaccharides showed increases in the numbers of carboxyl and carbonyl groups and double bonds. DPPH radical scavenging ability and reducing power of the gamma irradiated polysaccharides were significantly higher than those non-irradiated. PMID- 19285874 TI - Comparison of maximal elastance and systolic wall thickening using arterial tonometry and gated myocardial SPECT in patients undergoing coronary artery bypass grafting. AB - Myocardial SPECT using (99m)Tc-sestamibi, (99m)Tc-tetrofosmin, (201)Thallium is widely used in nuclear cardiology. Left ventricular systolic wall thickening (SWT) by SPECT and regional maximal elastance (rE(max)) using arterial tonometry were compared. rE(max) was calculated from time-pressure and time-volume curves. In normal heart, improvement of SWT was 4.1+/-11%, while 6.0+/-16% in dilated heart. Improvement of rE(max) was 0.67+/-1.0mmHg/mL in normal heart and 0.32+/ 0.7mmHg/mL in dilated heart (p<0.05). rE(max) can be an alternative variable as an index of regional contractility. PMID- 19285875 TI - Investigation on radiation degradation of carboxymethylcellulose by ionizing irradiation. AB - This study was done to compare the effects of irradiations with gamma-rays and electron beams, on the viscosity of the carboxymethylcellulose (CMC), on the functional groups of CMC, and on the production of radicals. It was observed that the relative viscosities decreased as the irradiation doses increased, but the decrease was more significant when irradiation with gamma rays. FT-IR spectra showed no significant difference between the gamma-ray and the electron beam irradiated samples. ESR spectra showed that the gamma-ray irradiation produced more radicals than electron beam irradiation in CMC. PMID- 19285876 TI - Effect of ionizing radiation on the physiological activities of ethanol extract from hizikia fusiformis cooking drips. AB - Although the byproduct from Hizikia fusiformis industry had many nutrients, it is being wasted. In this study, the physiological activities of cooking drip extracts from H. fusiformis (CDHF) were determined to investigate the effect of a gamma and an electron beam irradiations. DPPH radical scavenging activity and tyrosinase and ACE inhibition effects of the gamma and electron beam irradiated CDHF extracts were increased with increasing irradiation dose. These were reasoned by the increase in the content of the total polyphenolic compound of CDHF by the gamma and electron beam irradiation. There were no differences for the radiation types. These results show that ionizing radiation could be used for enhancing the functional activity of CDHF which is a major by-product in Hizikia fusiformis processing, in various applications. PMID- 19285877 TI - Accumulation and mobilization of arsenate by Fe(III) polyions trapped in a Ca polygalacturonate network. AB - The role of Fe(III) stored at the soil-root interface in the accumulation of arsenate and the influence of citric acid on the As(V) mobility were investigated by using Ca-polygalacturonate networks (PGA). The results indicate that in the 2.5-6.2 pH range Fe(III) interacts with As(V) leading to the sorption of As(V) on Fe(III) precipitates or Fe-As coprecipitates. The FT-IR analysis of these precipitates evidenced that the interaction produces Fe(III)-As(V) inner-sphere complexes with either monodentate or bidentate binuclear attachment of As(V) depending on pH. In the 3.0-6.0 pH range, As(V) diffuses freely through the polysaccharidic matrix that was found to exert a negligible reducing action towards As(V). At pH 6.0 citric acid is able to mobilize arsenate from the As-Fe PGA network through the complexation of the Fe(III) polyions that leads to the release of As(V). PMID- 19285878 TI - Ultraviolet radiation-A (366 nm) induced morphological and histological malformations during embryogenesis of Clarias gariepinus (Burchell, 1822). AB - Exposure to ultraviolet radiation has been associated with variety effects in many organisms ranging from molecular and tissue damage to population level effects. The exposure of embryos of the catfish, Clarias gariepinus (Burchell, 1822) to 366nm UVA at different doses 15, 30 and 60 min resulted in the hatching time delayed to 29 h-post-fertilization stage (29 h-PFS) in comparison with normal hatching time of 22h-PFS at 29 degrees C. In embryos exposed to 15 min/UVA, 30 min/UVA and 60 min/UVA the total percentage of hatched embryos/fertilized eggs were 90%, 89% and 85%, respectively, while in control was 95% at 29 h-PFS. The total percentage of mortality/ hatched embryos were (1-14)%, (2-22)%, (2-23)% and (3-40)% for control, 15 min, 30 min and 60 min groups, respectively, at 40 h-PFS. Also as a result some morphological malformations; (yolk sac oedema, body curvature, fin blistering, and dwarfism) were revealed. These destructive effects were also confirmed by histopathological changes in gills, eyes, intestinal tract, spinal cord, notochord, liver, skin and kidney. The results confirm that exposure to UVA caused an exposure time-dependent delay in hatching rate and reduced the percentage of the hatched embryos but the mortality rate increased with increase of the exposure time to UVA. PMID- 19285879 TI - Detection of UVBR-sensitive and -tolerant bacteria in surface waters of the western North Pacific. AB - In order to evaluate the effects of solar ultraviolet radiation (UVR) on eubacterial community composition, we examined the tolerance of eubacterial phylotypes to solar UV radiation in surface waters of the western North Pacific during September 2005. Bromodeoxyuridine (BrdU), a halogenated thymine analogue, was used for labeling newly synthesized DNA in proliferating cells. Thymine dimers (TD), which are specifically formed in DNA by biologically harmful ultraviolet B radiation (UVBR; 280-315nm), were also applied to detect UVB damaged genomes selectively. PCR-denaturing gradient gel electrophoresis (PCR DGGE) on the labeled samples revealed that UVBR-resistant cells showing active synthesis of DNA without accumulating TD, varied among phylotypes. In addition, UVBR-sensitive band positions with TD indicated inter-specific variations in sensitivity to UVBR. Phylogenetic analysis revealed that 12 DNA sequences were classified into eight phylogenetic groups: three Roseobacter, one Sphingomonas, two Gammaproteobacteria, one Actinobacteria, one Synechococcus, two Prochlorococcus, one plastid and one another group. A UVBR-resistant phylotype was affiliated to Erythrobacter sp. (previously designated as Sphingomonas sp.), which was distributed in warmer waters from the south of Oyashio to Kuroshio regions. A UVBR-sensitive phylotype was affiliated to Pseudoalteromonas sp. in Gammaproteobacteria. Dominant heterotrophic eubacteria were composed of both sensitive and resistant phylotypes. This is the first report on TD accumulated eubacterial phylotypes in oceanic surface waters. PMID- 19285880 TI - Comparative moleculo-immunological analysis of swamp- and riverine-type water buffaloes responses. AB - This moleculo-epidemiological and immunological study through cytokine response assessment was done to know the dynamics of cytokines in the initiation, persistence and association to physiological changes of a particular pathogen in water buffaloes. This is important to understand the magnitude and behavior of disease progression. Water buffalo blood samples gathered from different places in the Philippines revealed a 9.4%, 27.6%, 10.3% and 4.4% prevalence of bovine viral diarrhea virus (BVDV), bovine leukemia virus (BLV), Anaplasma marginale and Babesia bigemina infection, respectively. This was the first surveillance study of BVDV and BLV in the country. Furthermore, cytokine expression of these naturally infected animals was also quantified. BVDV-infected animals had up regulated expressions of TNFalpha, IL-2 and IL-4; and down-regulated expressions of IFNgamma and IL-12p40 while BLV positive animals had an up-regulated IL-4 and IL-6, and highly expressed IL-10 and IL-12p40 with unchanged IFNgamma expression. Meanwhile, animals infected with A. marginale had all interleukins and IFNgamma up-regulated with significant expression of IL-10 and IL-12p40 similar to the BLV positive animals. Since it was also observed that swamp-type buffaloes were more disease tolerant than riverine-type buffaloes based on the gathered infection rate of each examined pathogen, further assessment was done focusing on the two vital cytokines, IFNgamma and TNFalpha. We quantified IFNgamma and TNFalpha expressions in ConA-stimulated PBMC from both swamp and riverine buffaloes by real-time PCR. Cytokine expression from ConA-stimulated PBMC revealed that both IFNgamma and TNFalpha were more highly expressed in swamp than in riverine buffalo. To further examine the probable cause of expression differences, the proximal promoter region of these two cytokines were sequenced for the presence of nucleotide polymorphism followed by luciferase assay to analyze the effect of these polymorphisms in gene transcription. A single nucleotide polymorphism was found in the IFNgamma (-299) while eight polymorphisms in the TNFalpha promoter ( 541, -553, -562, -596, -609, -655, -659, -688). Luciferase assay showed that both IFNgamma promoter and TNFalpha promoter in swamp-type water buffalo had higher transcription activity compared to riverine-type water buffalo. These findings confirm that IFNgamma and TNFalpha transcriptions in these animals were highly affected by the disparity in the cytokine promoter region. This suggests that disease tolerance or susceptibility of these buffaloes could be due to the differences in their relative cytokine transcription and may relate to pathogen host specific pathogenesis. PMID- 19285881 TI - An experimental and theoretical investigation into the hydrolysis of dichloro(ethylenediamine)platinum(II) via electrospray mass spectrometry and density functional theory. AB - Dichloro(ethylenediamine)platinum(II), Pt(en)Cl(2), was dissolved in H(2)O and D(2)O, and the resulting aqueous solutions were electrosprayed into a quadrupole ion-trap mass spectrometer. A series of major and minor ionic hydrolysis products were detected. These ions were then subjected to collision-induced dissociation. As an aid in interpreting the experimental results, density functional theory calculations were carried out. These computations permitted the structures and energetics associated with the hydrolysis products to be determined. An understanding of the hydrolysis of PtenCl(2) and related coordination complexes is essential in the rational design of metal-based drugs. PMID- 19285882 TI - Issues associated with residual cell-substrate DNA in viral vaccines. AB - The presence of some residual cellular DNA derived from the production-cell substrate in viral vaccines is inevitable. Whether this DNA represents a safety concern, particularly if the cell substrate is derived from a tumor or is tumorigenic, is unknown. DNA has two biological activities that need to be considered. First, DNA can be oncogenic; second, DNA can be infectious. As part of our studies to assess the risk of residual cell-substrate DNA in viral vaccines, we have established assays that can quantify the biological activities of DNA. From data obtained using these assays, we have estimated the risk of an oncogenic or an infectious event from DNA. Because these estimates were derived from the most sensitive assays identified so far, they likely represent worst case estimates. In addition, methods that inactivate the biological activities of DNA can be assessed and estimations of risk reduction by these treatments can be made. In this paper, we discuss our approaches to address potential safety issues associated with residual cellular DNA from neoplastic cell substrates in viral vaccines, summarize the development of assays to quantify the oncogenic and infectivity activities of DNA, and discuss methods to reduce the biological activities of DNA. PMID- 19285884 TI - Understanding sleep-disordered breathing through mathematical modelling. AB - Recent studies have uncovered high prevalence of undiagnosed sleep-disordered breathing, and its linkage to metabolic or cardiovascular disorders which represent increasing health hazard. However, the mechanistic links behind these disorders as well as their contribution to the experimental observations and treatment responses remain poorly understood. Therefore, the screening of clinical measurements still relies upon relatively simple diagnostic features, such as signal averages or event frequencies, which may represent suboptimal or surrogate markers of the underlying abnormality. Consequently, most patients are being treated with general therapies regardless of the cause of their key dysfunction. Combining experimental measurements with mathematical modelling has the potential to provide mechanistic insights into the individual factors underlying the disease progression, which may finally enable tailored treatment alternatives for each patient. This review depicts a number of modelling approaches to elucidate sleep-related dysfunctions of the human respiratory system, and how these models are being used to translate the measurements first into new ideas and then into testable hypotheses. Such model-based investigations can provide systematic strategies towards better understanding, predicting or even preventing these dysfunctions. Along with the brief description of the modelling approaches, we discuss their relative merits and potential implications especially for clinical research. PMID- 19285885 TI - Biological and immunological characteristics of porcine follicle-stimulating hormone chemically modified with a polyethylene glycol derivative. AB - Porcine follicle-stimulating hormone (pFSH) was chemically modified with a polyethylene glycol (PEG) derivative to form PEGylated pFSH. The hormone was conjugated with a succinimidyl ester of methoxy PEG propionic acid (mPEG-SPA) at stoichiometric molar ratios of 1:0, 1:1, 1:3, 1:10, 1:30 and 1:100. The molecular weight of PEGylated pFSH increased depending on the stoichiometry of the conjugation reaction. Western blot analysis revealed a multifarious ladder pattern of PEGylated pFSH with apparent molecular weights of 31, 44, 66, 90 and 115 kDa. Differences between the immunopositive bands mostly coincided with the multiple of 5000 that was the molecular size of mPEG-SPA. PEGylated pFSH was biologically active and capable of stimulating both proliferation of and progesterone secretion by cultured bovine cumulus cells. PEGylated pFSH did not react with antibody against bovine FSH in a binding assay. The results indicate that PEGylated pFSH is still bioactive although this activity is reduced and the elimination of immunoreactivity by PEGylation suggests this modification may have merit in creating a product useful in inducing bovine superovulation. PMID- 19285886 TI - Chronic pain: avoidance or endurance? PMID- 19285887 TI - Improving postoperative follow-up of children receiving surgery for congenital or developmental cataracts in Africa. AB - PURPOSE: Access to cataract surgery for children has improved in recent years, and there are a number of tertiary facilities for children's eye health in sub Saharan Africa. However, surgery alone will have limited value if postoperative follow-up is poor. We have developed a program to improve follow-up after pediatric cataract surgery and compare the follow-up rates before and after implementation. METHODS: Baseline information was collected from 2003 to 2004. Program activities were implemented in 2005. Postintervention information was collected from 2006. During both periods all children younger than 16 who had cataract surgery at Kilimanjaro Christian Medical Centre Hospital were included. Follow-up, defined at both 2 weeks and 10 weeks, was recorded. RESULTS: Baseline data showed that 67% and 43% of children, respectively, came for 2-week and 10 week postoperative follow-up. After the intervention, 89% came for 2-week follow up and 83% came for 10-week follow-up. Gender inequity, documented in the baseline data, was eliminated in the postintervention period. CONCLUSION: Follow up of children who have had cataract surgery can be greatly enhanced in Africa. Improved follow-up enables children to receive the spectacles and low vision devices necessary for visual rehabilitation. Multiple strategies will probably be needed to improve follow-up. PMID- 19285888 TI - Recurrent isolated sixth nerve palsy after consecutive annual influenza vaccinations in a child. AB - Recurrent sixth nerve palsy in children in the absence of structural or other neurological abnormality is a rare occurrence. We report the case of recurrent isolated sixth (abducens) nerve palsy after consecutive annual influenza vaccinations in an otherwise-healthy 2-year-old boy. Investigations including magnetic resonance imaging of the brain and orbits after each episode failed to reveal any abnormality. The temporal relation to the immunizations supports but does not prove that the influenza immunization regimen was responsible. PMID- 19285889 TI - Visuospatial skills, ocular alignment, and magnetic resonance imaging findings in very low birth weight adolescents. AB - PURPOSE: To describe ocular alignment and stereoacuity in adolescents with very low birth weight (VLBW) in comparison with a matched control group and to investigate associations with white matter damage of immaturity (WMDI) and visuospatial skills in the VLBW group. METHODS: Fifty-nine 15-year-old VLBW subjects and 55 age- and sex-matched controls with normal birth weight underwent examination, including measurement of ocular alignment using cover test and Maddox rod and cycloplegic refraction. Stereoacuity was assessed with the TNO test, best-corrected visual acuity with a Konstantin Moutakis letter chart, and visuospatial skills with the performance tests, defined as performance intelligence quotient (IQ), in the Wechsler Intelligence Scale for Children (WISC III). All VLBW subjects underwent magnetic resonance imaging of the brain. RESULTS: Ocular misalignment was significantly more common in the VLBW group than in the control group (22% compared with 4%; p = 0.004). Exophoria, subnormal stereoacuity, and subnormal performance IQ were significantly more common in the VLBW group than in the control group (p = 0.006, p = 0.011, and p = 0.015, respectively). Ocular misalignment was associated with WMDI (p = 0.035) and subnormal performance IQ (p = 0.020). Of the VLBW subjects with ocular misalignment, 69% had WMDI and/or subnormal performance IQ. CONCLUSIONS: The VLBW adolescents had more visuospatial problems, lower stereoacuity, and more ocular misalignment than the control subjects. Ocular misalignment was associated with visuospatial deficiencies and/or WMDI in the VLBW group and was a better predictor for visuospatial deficits than WMDI. PMID- 19285890 TI - Intravitreal bevacizumab for idiopathic choroidal neovascularization. AB - An otherwise healthy 8-year-old girl with active juxtapapillary choroidal neovascularization was successfully treated with two intravitreal injections of bevacizumab. Intravitreal bevacizumab was well tolerated, choroidal neovascularization involuted, and subretinal and intraretinal serous fluid resorbed, with improvement in visual acuity from 20/400 to 20/50. The girl has remained stable for 6 months after the injections. PMID- 19285891 TI - Sympathetic ophthalmia secondary to cyclodiode laser in a 10-year-old boy. AB - A 10-year-old boy with tuberous sclerosis was treated with cyclodiode laser for right rubeotic glaucoma. Five months after treatment, he developed a bilateral granulomatous uveitis. Enucleation of the exciting eye demonstrated sympathetic ophthalmia. PMID- 19285892 TI - Position of helical kinks in membrane protein crystal structures and the accuracy of computational prediction. AB - The structural features of helical transmembrane (TM) proteins, such as helical kinks, tilts, and rotational orientations are important in modulation of their function and these structural features give rise to functional diversity in membrane proteins with similar topology. In particular, the helical kinks caused by breaking of the backbone hydrogen bonds lead to hinge bending flexibility in these helices. Therefore it is important to understand the nature of the helical kinks and to be able to reproduce these kinks in structural models of membrane proteins. We have analyzed the position and extent of helical kinks in the transmembrane helices of all the crystal structures of membrane proteins taken from the MPtopo database, which are about 405 individual helices of length between 19 and 35 residues. 44% of the crystal structures of TM helices showed a significant helical kink, and 35% of these kinks are caused by prolines. Many of the non-proline helical kinks are caused by other residues like Ser and Gly that are located at the center of helical kinks. The side chain of Ser makes a hydrogen bond with the main chain carbonyl of the i - 4th or i + 4th residue thus making a kink. We have also studied how well molecular dynamics (MD) simulations on isolated helices can reproduce the position of the helical kinks in TM helices. Such a method is useful for structure prediction of membrane proteins. We performed MD simulations, starting from a canonical helix for the 405 TM helices. 1 ns of MD simulation results show that we can reproduce about 79% of the proline kinks, only 59% of the vestigial proline kinks and 18% of the non proline helical kinks. We found that similar results can be obtained from choosing the lowest potential energy structure from the MD simulation. 4-14% more of the vestigial prolines were reproduced by replacing them with prolines before performing MD simulations, and changing the amino acid back to proline after the MD simulations. From these results we conclude that the position of the helical kinks is dependent on the TM sequence. However the extent of helical kinking may depend on the packing of the rest of the protein and the lipid bilayer. PMID- 19285893 TI - Contribution of fat-free mass and fat mass to bone mineral density among reproductive-aged women of white, black, and Hispanic race/ethnicity. AB - The objective of the study was to evaluate the contribution of fat-free mass (FFM) and fat mass (FM) to bone mineral density (BMD) and bone mineral apparent density (BMAD) among reproductive-aged women. Dual-energy X-ray absorptiometry scans were performed on 708 healthy black, white, and Hispanic women, 16-33 yr of age. The independent effect of FFM and FM on BMD and BMAD and the interaction of body composition measurements with race/ethnicity and age, were evaluated. FFM correlated more strongly than FM with BMD at the lumbar spine (r=0.52 vs r=0.39, p<0.01) and the femoral neck (r=0.54 vs r=0.41, p<0.01). There was a significant positive association between bone density measures [ln(BMD) and ln(BMAD)] and both ln(FFM) and ln(FM). The association of FFM with spinal BMD was stronger in 16-24-yr-old women than in 25-33-yr-old women (p<0.006). The effect of FFM on femoral neck BMD was greater in blacks (p<0.043) than Hispanics, whereas the effect of FM on spinal BMD was less (p<0.047). Both FM and FFM are important contributors to bone density although the balance of importance is slightly different between BMD and BMAD. PMID- 19285894 TI - Comparison of body composition by bioelectrical impedance and dual-energy x-ray absorptiometry in overweight/obese postmenopausal women. AB - The purpose of this study was to compare the Rudolph J. Liedtke (RJL) Sciences Quantum II system bioelectrical impedance analyzer (BIA) with the fan beam Hologic dual-energy X-ray absorptiometry (DXA, software V8.26a) for assessing body composition in postmenopausal obese women. Thirty-three postmenopausal overweight/obese females (mean age: 53.9+/-6.0 yr; mean weight: 91.3+/-17.5 kg; and mean body mass index [BMI]: 33.1+/-5.7 kg/m2) were evaluated for comparison of body weight (BW), fat mass (FM), percent FM (%FM), and fat free mass (FFM). The comparison was assessed by RJL Quantum 2 Cyprus 2.6 (Clinton Township, MI) BIA vs fan beam DXA Hologic QDR-4500A software V8.26a (ODR 4500 Hologic, Inc., Waltham, Mass). RJL-BIA and DXA measurements were performed at the same time. BW was measured using a balance scale (Detecto; Web City, MO) and these results were used for the RJL-BIA analysis. Balance weight was compared with DXA BW. Correlations between DXA and RJL-BIA for BW, FM, %FM, and FFM were 0.998, 0.980, 0.782, and 0.926 (p<0.01), respectively. Bland-Altman plots demonstrated general agreement between methods for BW, FM, %FM, and FFM. However, for the latter 3 metrics of body composition, one unit change using BIA does not correspond to one unit change using DXA, as there were systematic disagreements at either end of the range of values. But RJL-BIA could be a valid method for assessing body composition of overweight/obese postmenopausal women once appropriate validated regression equations have been developed. PMID- 19285895 TI - Observed occurrence of Tritrichomonas foetus and other enteric parasites in Australian cattery and shelter cats. AB - Cattery-housed pedigree cats, located mostly within the USA, have the highest reported prevalence of Tritrichomonas foetus (T foetus) to date. This prospective, multi-institutional, cross sectional study examines the occurrence of T foetus and other enteric parasites in cattery-housed and shelter cats within Australia, where T foetus has only recently been identified. Faecal specimens were collected from 134 cats, including 82 cattery-housed pedigree cats and 52 shelter cats. Faecal examinations performed for most cats included concentration techniques, Snap Giardia test, culture in InPouch medium, and polymerase chain reaction (PCR) amplification of T foetus ribosomal ribonucleic acid (rRNA) genes using species-specific primers. Observed occurrence of T foetus, Giardia species, Isospora species and Toxascaris leonina for cattery-housed cats (and catteries) were 0%, 7.4 (13.8)%, 10.9 (22.6)% and 1.6 (3.2)%, respectively. Observed occurrence of T foetus, Giardia species, Isospora species and hookworms for shelter cats were 0%, 11.5%, 9.8% and 4.9%, respectively. These results suggest the prevalence of T foetus in cattery-housed cats is currently much lower in Australia than in the USA, while Isospora and Giardia species infections are common. PMID- 19285896 TI - [Evaluation of an educational intervention in elderly patients with heart failure performed by nurses using a standardized care plan]. AB - OBJECTIVE: To determine the impact of an educational intervention after discharge in patients with heart failure (HF) performed by applying a standardized care plan centered on encouraging self-care and treatment adherence. MATERIAL AND METHODS: We performed a longitudinal study between January, 2006 and May, 2007 in a population of 112 patients selected after being included in a clinical pathway during their hospital stay. Five scheduled visits were made in the first 2 months after hospital discharge; during these visits, an educational care plan was applied to the patients individually and by means of an interview in the HF day hospital. The patients' level of knowledge was quantified by means of a 5-point Likert-like scale. The education was carefully evaluated at each visit. The results were expressed as means. To compare the variables, a repeated measures ANOVA was used. RESULTS: The mean age of the 100 patients who finalized both phases of the study was 78 years (SD: 7). The intervention was effective in evaluating the level of knowledge acquired by the patients about their disease (signs, symptoms, complications, etc.) and statistically significant differences were obtained in the education acquired at each visit and throughout the 60-day intervention. CONCLUSIONS: Health education in these patients and their families (main caregiver) improved their adaptation to their disease and increased autonomy and treatment adherence, thus improving quality of life. PMID- 19285897 TI - First-line anti-tuberculosis drug resistance patterns and trends at the national TB referral center in Iran--eight years of surveillance. AB - OBJECTIVE: Resistance to anti-tuberculosis (anti-TB) drugs is becoming a major and alarming threat in most regions worldwide. METHODS: This was a descriptive cross-sectional study at a tertiary hospital in Iran, using patient medical records for 2000-2003. The findings were analyzed following the same framework as that used for previous reports from this center. RESULTS: Among 1556 TB patients, drug susceptibility testing (DST) was performed for 548 culture-positive cases. Anti-TB drug resistance to both isoniazid and rifampin was identified in 10 (2.8%) of the new TB cases (multidrug-resistant TB; MDR-TB). Any resistance was detected in 228 (41.6%), showing an increasing trend in both new and retreatment cases. The data analysis revealed that drug-resistant TB had a statistically significant association with Afghan ethnicity, age>65 years, and the type of disease (retreatment vs. new TB case) (p<0.05). Also, assessment of the drug resistance trends showed a significant increase in resistance to any anti-TB agent, to isoniazid, and to streptomycin in new cases, and to all of the first line anti-TB drugs in retreatment patients. CONCLUSIONS: There has been an increasing trend in drug resistance in recent years, particularly in retreatment cases. Hence, revision of the national TB control program, reevaluation of the role of the World Health Organization category II (CAT II) regimen, as well as the conducting of a nationwide drug resistance survey, are recommended. PMID- 19285898 TI - Genetic profiling of Bolivian population using 15 STR markers of forensic importance. AB - Allele frequencies for the 13 CODIS markers plus D2S1358 and D19S433 loci were estimated in a sample of 148 unrelated individuals from Bolivia and parameters of forensic interest were calculated. Further, the STR data were analyzed using a distance-based method to assess the genetic relationships of this population with other ones living in Argentina, Brazil, Costa Rica, Mexico, Peru, Venezuela and three autochthonous populations living in the Beni Department of Bolivia (Quechua, Aymara and Beni population). PMID- 19285899 TI - Population data for 15 STR loci (Identifiler kit) in a Filipino population. AB - Allele frequencies for fifteen STR loci, D3S1358, TH01, D21S11, D18S51, D2S1338, D5S818, D13S317, D7S820, D16S539, CSF1PO, D19S433, vWA, D8S1179, TPOX and FGA, were investigated in a Filipino ethnic group resident in the United States and in the Philippines. Statistical evaluation of the data collected indicated the population to be in Hardy-Weinberg equilibrium and therefore acceptable for calculations in forensic and family relatedness casework. PMID- 19285900 TI - Alcohol study on blood concentration estimation: reliability and applicability of Widmark formula on Chinese male population. AB - Widmark formula suggests estimating blood alcohol concentration level (BAC) on a given amount of alcohol administrated with knowledge of subject's body weight and sex. The idea has been referenced extensively in forensic science with application in drink driving prosecution. A sample of Chinese subjects was collected in the drinking experiment, and in this study we mainly focus on the verification of the validity of Widmark formula on Chinese male population. A promising result is obtained where the extrapolated BAC measurement is directly proportional to the amount of alcohol administrated, and inversely to the subject's body weight. The results noted in the general linear model is further supported by the nonlinear regression analysis and a concordant argument reaches by deploying the concept to the BAC level attained at peak. PMID- 19285901 TI - Polymorphism studies of six miniSTR loci for three ethnic populations in Singapore. AB - MiniSTR loci have been demonstrated to be an effective approach in recovering genetic information from degraded specimen, because of the improved PCR efficiency of their reduced PCR amplicon sizes. This study constructed two miniSTR panels comprising six miniSTR loci, including D2S1776, D3S4529, D6S474, D9S2157, D10S1435 and D12ATA63. The allele frequency distribution, forensic parameters and heterozygosity in three Singapore populations are reported in this paper. All six loci showed relatively high polymorphism with observed heterozygosity > 0.7. However, many of these six loci also demonstrated significant population differentiation for the three populations. PMID- 19285902 TI - Allele frequencies of 15 autosomal STR loci in the southern Morocco population with phylogenetic structure among worldwide populations. AB - Polymerase chain reaction (PCR) amplification using the AmpFl STR Identifiler kit was performed in a random sample of 204 unrelated individuals from the Arabic speaking population of the southern Morocco. Allele frequencies of 15 STRs loci (D13S317, D16S539, D2S1338, vWA, TPOX, D18S51, D5S818, FGA, D8S1179, D21S11, D7S820, D19S433, CSF1PO, TH01 and D3S1358) have been reported in this population. Markers D18S51, FGA, D2S1338 and D21S11 had the highest power of discrimination (PD) values while TH01 was the most informative locus in the studied population. The phylogenetic tree established among worldwide populations and genetic distance values show a great affinity between the Southern Moroccan population, Saudian, Moroccan of Asni and Andalusian. Our data is useful for anthropological and other comparative studies of populations and is powerful for forensic and paternity testing in the Arabic-speaking population of the Southern Morocco. PMID- 19285903 TI - Forensic evaluation of STR data for the PowerPlex 16 System loci in a Bangladeshi population. AB - Allele frequencies of 15 autosomal STR loci included in PowerPlex 16 System were determined from a sample of 148 unrelated Bangladeshi individuals. Forensic efficiency parameters such as, the power of discrimination (PD), observed and expected heterozygosity (H), polymorphism information content (PIC), probability of match (PM), power of exclusion (PE), and typical paternity index were calculated for the loci. These parameters indicated the usefulness of the loci in paternity testing and personal identification in the Bangladeshi population. PMID- 19285904 TI - Environmental correlates of children's active transportation: a systematic literature review. AB - This systematic review investigated the environmental (physical, economic, socio cultural and political) correlates of active transportation (AT) among young people aged 5-18 years to better inform the promotion of active living. Greater distance, increasing household income and increasing car ownership are consistently associated with lower rates of AT among children. Having a non-white ethnic background has a convincing positive association with AT. Having recreation facilities and walk or bike paths present are possibly associated with higher rates of AT. Further research requires longitudinal and intervention studies, utilizing multi-level design methodologies and objective measures of environmental attributes. PMID- 19285905 TI - Auxin signaling in algal lineages: fact or myth? AB - Auxin is of major importance throughout the life cycle of a plant, affecting several physiological and developmental processes, such as cell expansion and division. However, the evolutionary time point at which auxin became involved in such diverse processes is currently unclear. Despite some controversy, numerous reports demonstrate the presence of auxin in algal lineages and its effects on algal development, suggesting an early evolutionary origin of auxin-dependent mechanisms. Here, we review these reports and discuss in silico analyses of auxin signaling components. It seems that, at least in microalgae, the assumed major components of auxin signaling in land plants are absent. However, these microalgae might have alternative auxin signaling pathways that could account for their responses to auxin. PMID- 19285906 TI - A morphogenetic trigger: is there an emerging concept in plant developmental biology? AB - Morphogens are involved in the establishment of positional information that is essential for pattern formation. In plants, the phytohormone auxin displays some characteristics of a morphogen. Gradients of auxin distribution are required for tissue patterning within the embryo and the root apex. In some other instances, such as de novo organogenesis, auxin action can be better described in terms of a morphogenetic trigger, which is defined as a factor that induces, through local increase of its concentration, acquisition of a new developmental fate in plant cells that were originally similar to their neighbours. A morphogenetic trigger specifies the site where a new organ will be formed. In plants, formation of reiterative and modular structures might need the action of both morphogenetic triggers and morphogens. PMID- 19285907 TI - Nuclear regulators with a second home in organelles. AB - In plants, increasing evidence points towards the existence of nuclear proteins that are also targeted to either mitochondria - a well-known phenomenon from yeast and mammalians - or to plastids. One such protein is Whirly1, which was the first protein to be identified in the nucleus and plastids of the same plant cell. Like Whirly1, most of the dual targeted (nucleus and organelle) proteins have functions in the maintenance of DNA, telomere structuring or gene expression. In some instances, proteins were even shown to be relocated from one compartment to another upon environmental or developmental clues. We hypothesize that one rationale of dual targeting is storage or sequestration of these proteins inside the organelles until specific conditions require their activity in the nucleus. PMID- 19285908 TI - Role of plant RNA-binding proteins in development, stress response and genome organization. AB - RNA-binding proteins (RBPs) in eukaryotes have crucial roles in all aspects of post-transcriptional gene regulation. They are important governors of diverse developmental processes by modulating expression of specific transcripts. The Arabidopsis (Arabidopsis thaliana) genome encodes for more than 200 different RBPs, most of which are plant specific and are therefore likely to perform plant specific functions. Indeed, recent identification and analysis of plant RBPs clearly showed that, in addition to the important role in diverse developmental processes, they are also involved in adaptation of plants to various environmental conditions. Clearly, they act by regulating pre-mRNA splicing, polyadenylation, RNA stability and RNA export, as well as by influencing chromatin modification. PMID- 19285909 TI - Thinking outside the F-box: novel ligands for novel receptors. AB - The importance of regulated proteolysis in the physiology and development of plants is highlighted by the large number of genes dedicated to proteasome dependent protein degradation. Within the SCF class of E3 ubiquitin ligases are more than 700 F-box proteins that act as recognition modules to specifically target their dedicated substrates for ubiquitylation. This review focuses on very recent studies indicating that some F-box proteins function as phytohormone or light receptors, which directly perceive signals and facilitate specific target protein degradation to regulate downstream pathways. If this new connection between ligand-regulated proteolysis and signaling proves to be more extensive, an entirely new way of understanding the control of signal transduction is in the offing. PMID- 19285910 TI - Automated model-based vertebra detection, identification, and segmentation in CT images. AB - For many orthopaedic, neurological, and oncological applications, an exact segmentation of the vertebral column including an identification of each vertebra is essential. However, although bony structures show high contrast in CT images, the segmentation and labelling of individual vertebrae is challenging. In this paper, we present a comprehensive solution for automatically detecting, identifying, and segmenting vertebrae in CT images. A framework has been designed that takes an arbitrary CT image, e.g., head-neck, thorax, lumbar, or whole spine, as input and provides a segmentation in form of labelled triangulated vertebra surface models. In order to obtain a robust processing chain, profound prior knowledge is applied through the use of various kinds of models covering shape, gradient, and appearance information. The framework has been tested on 64 CT images even including pathologies. In 56 cases, it was successfully applied resulting in a final mean point-to-surface segmentation error of 1.12+/-1.04mm. One key issue is a reliable identification of vertebrae. For a single vertebra, we achieve an identification success of more than 70%. Increasing the number of available vertebrae leads to an increase in the identification rate reaching 100% if 16 or more vertebrae are shown in the image. PMID- 19285911 TI - Interference: unique source of forgetting in working memory? PMID- 19285912 TI - Natural pedagogy. AB - We propose that human communication is specifically adapted to allow the transmission of generic knowledge between individuals. Such a communication system, which we call 'natural pedagogy', enables fast and efficient social learning of cognitively opaque cultural knowledge that would be hard to acquire relying on purely observational learning mechanisms alone. We argue that human infants are prepared to be at the receptive side of natural pedagogy (i) by being sensitive to ostensive signals that indicate that they are being addressed by communication, (ii) by developing referential expectations in ostensive contexts and (iii) by being biased to interpret ostensive-referential communication as conveying information that is kind-relevant and generalizable. PMID- 19285913 TI - How do emotion and motivation direct executive control? AB - Emotion and motivation have crucial roles in determining human behavior. Yet, how they interact with cognitive control functions is less understood. Here, the basic elements of a conceptual framework for understanding how they interact are introduced. More broadly, the 'dual competition' framework proposes that emotion and motivation affect both perceptual and executive competition. In particular, the anterior cingulate cortex is hypothesized to be engaged in attentional/effortful control mechanisms and to interact with several other brain structures, including the amygdala and nucleus accumbens, in integrating affectively significant signals with control signals in prefrontal cortex. An implication of the proposal is that emotion and motivation can either enhance or impair behavioral performance depending on how they interact with control functions. PMID- 19285915 TI - Resistance to antimalarial compounds: methods and applications. AB - Considerable progress has been made in antimalarial research; however, much more effort is required to develop new antimalarials which overcome plasmodial resistance to clinically available drugs and limit the emergence of resistance. In particular, research must focus on understanding the mechanisms by which Plasmodium falciparum impedes the action of these drugs. In this review, we discuss the different in vitro and in vivo models used to adequately study resistance to antimalarial compounds. For each molecule studied, the best in vitro and/or in vivo model is highlighted with a detailed discussion of its validity in relation to field data on P. falciparum resistance. PMID- 19285916 TI - Bis(azido) alpha-iminooxime cobalt macrocyclic complex: synthesis, spectroscopy and ZINDO/S calculations. AB - Herein we report on the macrocyclic complex [Co(dohpn)(N3)2] (dohpn=2,3,9,10 tetramethyl-1,4,8,11-tetraazaundecane-1,3,8,10-tetraen-11-ol-1-olate). According to the MM+ optimized structure, it shows an octahedral environment around the metal ion with an overall planar configuration of the macrocyclic ligand, while azido ligands are not perpendicular to the plane of the macrocycle. Quantum mechanic calculations through semi-empirical methods were performed and a detailed spectroscopic (ZINDO/S) assignment is also presented. PMID- 19285917 TI - Spectroscopic and DFT studies of flurbiprofen as dimer and its Cu(II) and Hg(II) complexes. AB - The vibrational study in the solid state of flurbiprofen and its Cu(II) and Hg(II) complexes was performed by IR and Raman spectroscopy. The changes observed between the IR and Raman spectra of the ligand and of the complexes allowed us to establish the coordination mode of the metal in both complexes. The comparative vibrational analysis of the free ligand and its complexes gave evidence that flurbiprofen binds metal (II) through the carboxylate oxygen. The fully optimized equilibrium structure of flurbiprofen and its metal complexes was obtained by density functional B3LYP method by using LanL2DZ and 6-31 G(d,p) basis sets. The harmonic vibrational frequencies, infrared intensities and Raman scattering activities of flurbiprofen were calculated by density functional B3LYP methods by using 6-31G(d,p) basis set. The scaled theoretical wavenumbers showed very good agreement with the experimental values. The electronic properties of the free molecule and its complexes were also performed at B3LYP/6-31G(d,p) level of theory. Detailed interpretations of the infrared and Raman spectra of flurbiprofen are reported. The UV-vis spectra of flurbiprofen and its metal complexes were also investigated in organic solvents. PMID- 19285918 TI - Safety study of 50 Hz repetitive transcranial magnetic stimulation in patients with Parkinson's disease. AB - OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) has shown promising results in treating Parkinson's disease (PD), but the best values for rTMS parameters are not established. Fifty Hertz rTMS may be superior to 25 Hz rTMS investigated so far. The objective of this study was to determine if 50 Hz rTMS could be delivered safely in PD patients since current safety limits are exceeded. METHODS: Fifty Hertz rTMS was applied with a circular coil on the primary motor cortex (M1). Stimulation intensity was first tested at 60% rest motor threshold [RMT] and 0.5 s train duration and then increased in 0.5 s steps to 2 s, and by 10% steps to 90% RMT. Multi-channel electromyography (EMG) was recorded to control for signs of increasing time-locked EMG activity including correlates of the spread of excitation and after-discharges, or an increase of M1 excitability. Pre- and post-50 Hz rTMS assessments included EEG, Unified Parkinson Disease Rating Scale (UPDRS), Grooved Pegboard Test, Serial Reaction Time Task (SRTT), Folstein Mini-Mental Status Examination (MMSE) and Verbal Fluency to control for motor and cognitive side effects. RESULTS: Ten PD patients were investigated. Multi-channel EMG showed no signs of increased time-locked EMG activity including correlates of the spread of excitation and after-discharges, or increased M1 excitability in 9 patients. A PD patient with bi-temporal spikes in the pre-testing EEG had clinical and EMG correlates of spread of excitation at 90% RMT, but no seizure activity. Pre- and post-50 Hz assessment showed no changes. No adverse events were observed. Fifty Hertz rTMS was well tolerated except by 1 patient who wished to terminate the study due to facial muscle stimulation. CONCLUSION: Fifty Hertz rTMS at an intensity of 90% RMT for 2 s appears safe in patients with PD, but caution should be taken for patients with paroxysmal EEG activity. For this reason, comprehensive screening should include EEG before higher-frequency rTMS is applied. SIGNIFICANCE: This is the first study to investigate safety of 50 Hz rTMS in humans. PMID- 19285919 TI - Purinergic receptors in airway epithelia. AB - Nucleotides and nucleosides within the airway surface liquid regulate mucociliary clearance (MCC) activities, the primary innate defense mechanism that removes foreign particles and pathogens from airway surfaces. Nucleotide and nucleoside actions in the airways are mediated mainly by two purinergic receptor subtypes, the Gq-coupled ATP/UTP-sensing P2Y2 receptor and the Gs-coupled A2b adenosine receptor. Activation of the A2b receptor results in cyclic AMP-dependent activation of the cystic fibrosis transmembrane regulator (CFTR) Cl- channel and stimulation of ciliary beat frequency. Agonist occupation of the P2Y2 receptor promotes inhibition of Na+ absorption as well as CFTR-dependent and CFTR independent Cl- secretion, ciliary beating, and mucin secretion. PMID- 19285920 TI - Increasing hazard of Sprint Fidelis implantable cardioverter-defibrillator lead failure. AB - BACKGROUND: The Medtronic Sprint Fidelis defibrillator lead is prone to fracture and was recalled in 2007 after 665 failures and five reported deaths. Approximately 150,000 patients at risk for sudden death in the United States have Sprint Fidelis leads. The rate of Sprint Fidelis lead failure may be increasing, and physicians are confronted with the decision to replace the lead prophylactically. OBJECTIVE: The purpose of this study was to determine if the hazard of Sprint Fidelis lead failure is changing and to compare its performance to other contemporary implantable cardioverter-defibrillator (ICD) leads. METHODS: Transvenous ICD leads implanted and followed at our two tertiary-care referral centers between January 2004 and December 2008 were included in the study. Lead failure data were entered prospectively by both centers via the Multicenter Registry. Clinical data were collected prospectively by each center and merged for the purpose of this study. RESULTS: During 5,700 implant years of follow-up (average 1.9 +/- 1.3 years), 94 of 3,037 defibrillator leads failed (1.65%/year), including 72 (8.5%) of 848 Sprint Fidelis leads. The cumulative hazard of Sprint Fidelis failure was significantly greater compared to 2,189 other defibrillator leads (P <.0001), and the hazard of Sprint Fidelis failure accelerated after the first year and continued to increase during the study. In contrast to other defibrillator leads, the Sprint Fidelis failure rate was significantly higher (3.75%/year vs 0.58%/year) and the 3-year estimated survival was significantly lower (87.9% [95% CI 84.8, 90.9] vs 98.5% [95% CI 97.8, 99.3]; P <.0001). The chance that a Sprint Fidelis lead would survive another year decreased progressively during the study. Most Sprint Fidelis failures were caused by pace-sense conductor fracture (n = 63 [87.5%]), which caused inappropriate shocks in 36 of 72 patients. CONCLUSION: The hazard of Sprint Fidelis lead failure is increasing, whereas the failure rates of other defibrillator leads are low and stable. Physicians should consider these data when managing patients who have Sprint Fidelis leads. PMID- 19285921 TI - Vaginal vault dehiscence after hysterectomy. AB - STUDY OBJECTIVE: The purpose of our study was to evaluate factors predisposing vault dehiscence after hysterectomy and its manifestation. DESIGN: Case series and review of the literature (Canadian Task Force classification II-3). SETTING: Multicenter study. PATIENTS: Retrospective analysis of 16 unpublished cases of vaginal vault dehiscence after total laparoscopic hysterectomy from physicians who participated in the exchange on the topic of vaginal vault dehiscence at the American Association of Gynecologic Laparoscopists Endo Exchange List (group A) and review of 38 reported cases in the literature (group B). INTERVENTIONS: The participating physicians were asked to complete a detailed questionnaire related to vault dehiscence. In addition, we performed literature search using the keywords "vault dehiscence," "vaginal vault dehiscence," "vault prolapse," and "hysterectomy," and conducted the search in MEDLINE, EMBASE, and Cochrane Database of Systematic Reviews. MEASUREMENTS AND MAIN RESULTS: We estimated risk factors and characteristic features for vaginal vault dehiscence. The incidence of vault dehiscence was higher after laparoscopic hysterectomy (1.14%) than after abdominal hysterectomy (0.10%, p <.0001, OR 11.5) and after vaginal hysterectomy (0.14%, p <.001, OR 8.3). The time interval between hysterectomy and occurrence of vault dehiscence in the laparoscopic group (8.4 +/- 1.2 weeks) was significantly shorter than in the abdominal hysterectomy (112.7 +/- 75.1 weeks, p = .01) and in vaginal hysterectomy (136.5 +/- 32.2 weeks, p <.0001) groups, respectively. It appears that sexual intercourse was the main triggering event for vault dehiscence (58.8%). Vaginal bleeding (50%) and vaginal evisceration (48.1%) were the main symptoms. CONCLUSION: Our data suggest that vaginal vault dehiscence is rare but may occur more often after laparoscopic hysterectomy than after other hysterectomy approaches. Whether it is related to the technique of laparoscopic suturing is unclear. Other risk factors such as early resumption of regular activities and sexual intercourse may play a role. PMID- 19285922 TI - Effect of upper abdomen tissue manipulation on adhesion formation between injured areas in a laparoscopic mouse model. AB - STUDY OBJECTIVE: These experiments were designed to examine the effect of manipulation during surgery as a cofactor in adhesion formation at trauma sites. DESIGN: Randomized, controlled trial. Canadian Task Force Classification-class 1. SETTING: University laboratory research center. SUBJECTS: A standardized laparoscopic mouse model (Balb?c mice 9-10 weeks old) for adhesion formation after opposing bipolar lesions and 60 minutes of carbon-dioxide pneumoperitoneum. In this model adhesions are known to decrease after the addition of 3% of oxygen, dexamethasone, or both. In addition, adhesions decrease with experience (i.e., with a decreasing amount of manipulation during the learning curve). INTERVENTIONS: A factorial design was used to evaluate the effects of dexamethasone and of adding 3% of oxygen on manipulation-enhanced adhesion formation during a learning curve. Blocks of 4 animals were thus randomized as controls (carbon-dioxide pneumoperitoneum only) or received an additional 3% of oxygen, dexamethasone, or both. In a second experiment, the effects of manipulation on adhesion formation were quantified. In a third experiment we evaluated whether dexamethasone had a specific effect on manipulation-enhanced adhesion formation. MEASUREMENTS AND MAIN RESULTS: Qualitative and quantitative adhesion scoring 7 days after the intervention. The first experiment confirmed that adhesion formation decreased during the learning curve (p <.0001) and after the addition of dexamethasone whether assessed as the total adhesion score (p <.0001 and p =.0009, respectively) or a quantitative score (p <.0001 and p <.0001, respectively). The second experiment showed that adhesion formation increased by standardized touching and grasping of omentum and bowels (proportion score p =.0059 and p =.0003, respectively) and this effect increased with duration of touching (p =.0301). In the third experiment, dexamethasone was confirmed to decreased adhesion formation (p =.0001) but this effect was not specific for manipulation-enhanced adhesion formation. CONCLUSION: Manipulation of intraperitoneal organs in the upper abdomen enhances adhesion formation at trauma sites, confirming that the peritoneal cavity is a cofactor in adhesion formation. Dexamethasone decreases adhesion formation but the effect is not specific for manipulation-enhanced adhesion formation. PMID- 19285923 TI - Determination of duloxetine in human plasma by capillary electrophoresis with laser-induced fluorescence detection. AB - A method based on capillary electrophoresis has been developed for the analysis of the novel antidepressant drug duloxetine in human plasma. The method makes use of laser-induced fluorescence detection after derivatisation of the analyte with 5-(4,6-dichlorotriazinyl)aminofluorescein at pH 11. A single step liquid/liquid extraction procedure with a mixture of hexane/2-propanol allows the sample clean up with extraction yields always >or=84% and interference removal. The electrophoretic separation is achieved using uncoated fused silica capillaries (60.0 cm effective length, 75.0 cm total length, 50 microm internal diameter) and a background electrolyte composed of borate buffer (40 mM, pH 10.3), tetrabutylammonium bromide (10 mM), and acetone (10%, v/v). The applied voltage is 20 kV; the samples are injected by pressure (50 mbar x 8 s). The method has been fully validated in terms of linearity range (2.5-150 ng mL(-1)), LOD and LOQ (1.0 and 2.5 ng mL(-1), respectively), precision (R.S.D.<6.7%) and accuracy (recovery >78%). Application to samples obtained from patients under treatment with duloxetine gave good results. The method represents the first application of capillary electrophoresis to the analysis of duloxetine in human plasma. PMID- 19285924 TI - Quantitative analysis of sphingosine-1-phosphate by HPLC after napthalene-2,3 dicarboxaldehyde (NDA) derivatization. AB - Sphingosine-1-phosphate (S1P) is an important sphingolipid signaling molecule that regulates numerous cellular processes. In this paper we report a new method to quantify the levels of S1P in biological samples that relies on derivatization with naphthalene-2,3-dicarboxaldehyde (NDA) and quantification by reverse-phase high performance liquid chromatography (HPLC). The limit of detection (LOD) using S1P standards was 20.9fmol (12.6nM), and the limit of quantification (LOQ) was 69.6fmol (41.7nM). The recovery of S1P standards was up to 97.5%. The mean relative standard deviations (RSD) for the intra- and inter-day assay were 4.1% and 7.7%, respectively. To validate this procedure, we quantified the S1P levels in plasma from human, horse, and mouse (mean values of 0.45, 0.25, and 1.23microM, respectively). We also used this technique to evaluate the S1P content in mouse tissues, as well as in rat neuronal cell cultures before and after sphingosine treatment. The results demonstrate that this new procedure can provide fast, sensitive, and reproducible S1P quantification, and offers several advantages over existing methods. The technique also may be used for determining the activity, as well as the inhibition, of sphingosine kinase. In the future it could be an important tool for investigators studying the role of S1P in signal transduction, cell growth and differentiation, and disease pathogenesis and treatment. PMID- 19285925 TI - Determination of lumefantrine in rat plasma by liquid-liquid extraction using LC MS/MS with electrospray ionization: assay development, validation and application to a pharmacokinetic study. AB - A simple, sensitive and rapid method for the analysis of lumefantrine in rat plasma using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed. Detection was performed by positive ion electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The method included a chromatographic run of 5 min using a C(18) analytical column and the calibration curve was linear over the concentration range of 2-500 ng/mL with a correlation coefficient (r) of 0.996 or better. The intra- and inter-day assay precision ranged from 1.5 to 7.5% and 5.5 to 7.7%, respectively, and intra- and inter-day assay accuracy was between 91.3-109.7% and 97.0-104.7%, respectively. The method was successfully applied for the pharmacokinetic study in rats. PMID- 19285926 TI - Quantification of peramivir (a novel anti-influenza drug) in human plasma by hydrophilic interaction chromatography/tandem mass spectrometry. AB - Peramivir is a novel influenza neuraminidase inhibitor. In this article, hydrophilic interaction chromatography coupled with tandem mass spectrometry was developed to determine peramivir in human plasma. The positive ion MRM mode was performed and the precursor to the product ion transitions of m/z 329-->100 and 285-->138 were used to measure peramivir and Ro 64-0802 (I.S.). Chromatographic separation was performed on an Amide-80 column with acetonitrile-water-formic acid (70:30:0.1, v/v/v, 0.5mL/min). The method was linear over a concentration range of 10-10,000ng/mL. The average inter-day/intra-day precision values were 3.7+/-1.8% and 4.3+/-1.8%, respectively, while the accuracy values were 97.0+/ 4.8%. This method has been successfully applied to Phase I of clinical research of peramivir. PMID- 19285927 TI - Development of a sensitive and selective LC-MS/MS method for the determination of alpha-fluoro-beta-alanine, 5-fluorouracil and capecitabine in human plasma. AB - A sensitive and selective quantitative method to determine alpha-fluoro-beta alanine (FBAL), 5-fluorouracil (5-FU), and capecitabine (Cape) from a single human plasma aliquot (50 microL) has been developed and validated. First, 5-FU and Cape were extracted by liquid-liquid extraction (LLE) using a mixture of acetonitrile and ethyl acetate. This was followed by derivatization with dansyl chloride. The dansyl-derivatives from 5-FU and Cape were further purified using LLE with methyl tertiary-butyl ether (MTBE) and analyzed using a reversed-phase analytical column "Primesep D" (2.1 mm x 50 mm; 5 microm) with embedded basic ion pairing groups. The remaining aqueous phase containing FBAL was treated with dansyl chloride and the dansyl-FBAL was purified by solid phase extraction. Ultra high pressure liquid chromatography (UPLC) technology on a BEH C18 stationary phase column with 1.7 microm particle size was used for analysis of dansyl-FBAL. The method was validated over the concentration ranges of 10-10,000, 5-5000, and 1-1000 ng/mL for FBAL, 5-FU, and Cape, respectively. The results from assay validation show that the method is rugged, precise, accurate, and well suited to support pharmacokinetic studies where approximately 300 samples can be extracted and analyzed in 1 day. PMID- 19285928 TI - Determination of lincomycin in urine and some foodstuffs by flow injection analysis coupled with liquid chromatography and electrochemical detection with a preanodized screen-printed carbon electrode. AB - An electroanalytical method for the determination of lincomycin in feeds, honey, milk and urine was demonstrated in this study. The procedure employed a solid phase extraction for the isolation of lincomycin from real samples. The antibiotic residues were subsequently analyzed by high-performance liquid chromatography (HPLC) coupled with a disposable electrochemical sensor. The use of a disposable sensor together with the application of solid-phase extraction is attractive in practical application and should be useful in fast screening assay. The electroanalysis of lincomycin was first investigated using a preanodized screen-printed carbon electrode (SPCE*). Note that the SPCE* holds the advantages of low cost and easy to handle. The analytical parameters, such as, preanodization potential, preanodization time, solution pH, detection potential, cartridge, wash solution, elute solution and mobile phase, were further studied in detail. Under optimized conditions, the linear detection range for lincomycin is up to 1mM (correlation coefficient=0.999) with a detection limit of 0.08microM (S/N=3) and a quantification limit of 0.27microM (S/N=10). The applicability of the method was successfully demonstrated in real sample analysis. PMID- 19285929 TI - Protein supply and nutritional status in nineteenth century Bavaria, Prussia and France. AB - What determined regional height differences in the 19th century? We compare anthropometric evidence with production estimates of different food products and other economic variables. To this end, we concentrate on 179 rural regions and 29 towns in Bavaria (Southeast Germany). This regionally disaggregated level of analysis enables us to study the influence of the local supply of different food products on the nutritional status of the population, among which milk turned out particularly important. This result is tested and confirmed with regional data from Prussia and France. PMID- 19285930 TI - Porous silicon as a potential electrode material in a nerve repair setting: Tissue reactions. AB - We compared porous silicon (pSi) with smooth Si as chip-implant surfaces in a nerve regeneration setting. Silicon chips can be used for recording neural activity and are potential nerve interface devices. A silicon chip with one smooth and one porous side inserted into a tube was used to bridge a 5 mm defect in rat sciatic nerve. Six or 12 weeks later, new nerve structures surrounded by a perineurium-like capsule had formed on each side of the chip. The number of regenerated nerve fibers did not differ on either side of the chip as shown by immunostaining for neurofilaments. However, the capsule that had formed in contact with the chip was significantly thinner on the porous side than on the smooth side. Cellular protrusions had formed on the pSi side and the regenerated nerve tissue was found to attach firmly to this surface, while the tissue was hardly attached to the smooth silicon surface. We conclude that a pSi surface, due to its large surface area, diminished inflammatory response and firm adhesion to the tissue, should be a good material for the development of new implantable electronic nerve devices. PMID- 19285931 TI - From research to clinical practice an interdisciplinary project of day surgery anaesthesiological course: from preoperative evaluation to patient discharge. AB - This paper describes the anaesthesiological course of patients cared by an independent multi skills Day Surgery Unit during four years activity from 2004 to 2007. This method of caring surgical patients, continuously expanding, calls for regular audits where the anaesthetist plays a major role in selecting patients, in choosing and applying the most suitable anaesthesia technique, in managing any postoperative complications, and discharging the patient. All this contributes to minimising the patient's discomfort, to a quick recovery, and to minimise all inconvenience that may invalidate the service provided. Our experience includes 13,014 surgical procedures performed by seven specialised Operative Units. The analysis of our case records shows we have established an adequate organisation model and achieved a high quality standard of care in connection with high volumes of activity. PMID- 19285932 TI - Tranquilli-Leali or Atasoy flap: an anatomical cadaveric study. AB - BACKGROUND: The Tranquilli-Leali and Atasoy volar V-Y advancement flaps are considered workhorse flaps in the reconstruction of fingertip amputations. However, their description in the literature in terms of surgical dissection and blood supply is often indistinct. This study describes the differences between the two flaps and highlights their unique blood supply based on a thorough cadaveric study and review of the literature. METHODS: Using 16 fresh cadaveric fingers, eight Tranquilli-Leali and eight Atasoy volar V-Y advancement flaps were dissected, mapping the arterial blood supply using an injectable blue resin. In addition, a thorough literature search on the subject was done. RESULTS: In all eight fingertips dissected as decribed by Tranquilli-Leali, the flap was supplied by the anastomotic connections between the terminal branches of the palmar digital arteries and dorsal nail-bed arcades via the fibro-osseous hiatus. In contrast, in all eight fingertips which were dissected as described by Atasoy, the flaps were perfused through the terminal branches of the palmar digital arteries. CONCLUSIONS: The Tranquilli-Leali and Atasoy volar V-Y advancement flaps, used to reconstruct fingertip amputations, are distinct from one another in several ways. The most obvious difference is their technique of flap dissection, which, in turn, dictates a unique blood supply. Through careful dissection and a review of the literature, this anatomical study has brought to light the specific vascular supply to each flap that was evaluated. PMID- 19285933 TI - Self-titration of biphasic insulin aspart 30/70 improves glycaemic control and allows easy intensification in a Dutch clinical practice. AB - AIMS: This 18-month study assessed the improvement in glycaemic control and proportion of patients reaching glycated haemoglobin (HbA(1c)) targets with biphasic insulin aspart 30/70 (BIAsp 30) in clinical practice. METHODS: Type-2 diabetes patients failing on oral antidiabetic drugs (n=90) or existing insulin regimens (n=59) started or switched to BIAsp 30. Thiazolidinediones were stopped, metformin was continued. BIAsp 30 was given once daily (n=41), twice daily (n=96), or three times daily (n=12). Patients were taught self-monitoring and self-titration using an algorithm, adding daily doses of BIAsp 30 when necessary. RESULTS: Mean baseline HbA(1c) was 8.4%, weight 85.4 kg, and age 57.9 years. All patients experienced significant reductions in HbA(1c) (mean 1.9%+/-0.1), fasting plasma glucose (mean 2.8 mmol/l), and post-prandial glycaemia (mean 2.9 mmol/l); 91% of patients achieved HbA(1c)<7% and 52% achieved HbA(1c) < or=6.5%. No major or nocturnal hypoglycaemia were reported; 15% of patients reported minor hypoglycaemia. Insulin-naive patients gained mean 2.7 kg; patients who switched from another insulin lost weight (mean -0.6kg). CONCLUSION: The results from this study from routine care suggest that BIAsp 30 may allow a large proportion of type-2 diabetes patients (90%) to improve glycaemic control and reach target HbA(1c)<7%, using self-titration. PMID- 19285934 TI - The evolution of the natriuretic peptides - Current applications in human and animal medicine. AB - Although natriuretic peptides have played an important role in the fluid homeostasis of vertebrates for over several million years, their importance has only been noticed in the last few decades. Yet, the family of natriuretic peptides have since their discovery, drawn the attention of a broad spectrum of physicians and researchers involved in the maintenance of fluid homeostasis, including marine biologists, basic scientists, physicians and veterinarians. While all natriuretic peptides share a common phylogenetic background, due to differences in receptor-binding affinities, they have evolved into different hormones with clear distinct functions. B-type natriuretic peptide (BNP) is the most studied member of the natriuretic peptide family, and together with its cleavage equivalent amino-terminal proB-type natriuretic peptide (NT-proBNP) these peptides have emerged as important cardiovascular serum markers. However, since their introduction, physicians involved in human or animal medicine have faced common but also different challenges in order to optimally interpret the diagnostic and prognostic value of these novel cardiovascular biomarkers. PMID- 19285935 TI - Midwives' emotional wellbeing: impact of conducting a structured antenatal psychosocial assessment (SAPSA). AB - RESEARCH PROBLEM: To investigate the impact of conducting structured antenatal psychosocial assessments (SAPSA) on midwives' emotional wellbeing. The SAPSA includes screening and assessment tools for domestic violence, childhood trauma, drug and alcohol use, depression, and vulnerability factors. PARTICIPANTS AND METHODS: Registered midwives who had conducted the SAPSA with women during the first hospital booking visit at two hospitals in NSW. Data was collected by means of focus group interviews. RESULTS: Four sub-themes were identified that directly impacted upon the midwives' emotional wellbeing: cumulative complex disclosures, frustration and stress, lack of support for midwives and unhealthy coping strategies. DISCUSSION AND CONCLUSIONS: There was a cumulative emotional effect with some midwives utilising unhealthy strategies to cope with feelings of frustration, inadequacy and vicarious trauma. Establishment of structured referral pathways for women and supportive systems for midwives is essential prior to implementing the SAPSA. PMID- 19285936 TI - Sickle-cell hip necrosis and intraosseous pressure. AB - INTRODUCTION: Osteonecrosis of the femoral head (ONFH) is a frequent complication of sickle-cell disease. Numerous studies have demonstrated increased intraosseous pressure (IOP) in idiopathic necrosis and necrosis secondary to corticotherapy or alcohol poisoning. Several reports have testified to the clinical interest of decompression by drilling which, when performed in the early course of the pathology, can arrest or slow evolution. To the best of our knowledge, no studies have reported IOP increase in sickle-cell ONFH. The present study sought to show that intraosseous hyperpressure plays a role in the physiopathology of sickle cell, like idiopathic, ONFH. MATERIALS AND METHODS: Sixteen intraosseous pressure (IOP) measurements were taken: eight in adult sickle-cell disease patients, four in sickle-cell trait carrying ONFH patients (AS) and four in non-sickle-cell ONFH patients (AA). Arterial blood-pressure equipment with bone-puncture needle was used to measure IOP in the great trochanter body. Three IOP measurements were made after zero calibration: before drilling (direct pressure: IOP-1), after hyperpressure test but before drilling (IOP-2), and after drilling (IOP-3). RESULTS: The present, admittedly short, series displayed elevated predrilling IOP 1 and IOP-2, reduced after drilling (IOP-3). Abnormal IOP fell after drilling performed for evolutive symptomatic ONFH. Significant differences in IOP-1 and IOP-2 were found, these being higher in the "sickle-cell disease" and "sickle cell trait carriers" groups (p<0.05). Only in the sickle-cell groups was there a significant correlation between pain score and hyperpressure level, with significantly reduced pain after drilling. DISCUSSION: The elevated IOP levels found in symptomatic sickle-cell hips were comparable to those reported in the literature. Ischemia due to femoral head sinusoid occlusion by falciform globules with secondary intraosseous hyperpressure is the cause of the pain and of the onset and evolution of ONFH. The drilling tunnel acts as a safety valve, achieving real decompression of the segment involved and immediate postoperative reduction in or disappearance of pain. CONCLUSION: Measuring pressure is of diagnostic interest in sickle-cell disease patients with symptomatic hips. Manometry can be performed independently of surgery, under local anesthesia, and provides early confirmation of ONFH in geographic regions in which MRI is not readily available. It can be carried out very straightforwardly, without pressure sensor, using a simple water column (physiological saline) and three-way tap. Peroperative comparison of IOP-1 and IOP-3 is a means of assessing the effectiveness of decompression drilling. PMID- 19285937 TI - Dangerous feather! AB - A 57-year-old man presented to an outside hospital with episodes of persistent fever. On further history, the patient stated that his current condition started when he "stuck" himself in the back with a "sharp feather" protruding from his sofa 2 weeks before. He underwent transesopheageal echocardiography (TEE) to assess for endocarditis. TEE showed no evidence of endocarditis; however, the midesophageal TEE view showed a large mobile mass in the long axis view of the descending thoracic aorta. On CT imaging of the thoracic aorta, the large mass with attachment just distal to the left subclavian artery was noted. The decision was made that this friable and highly mobile mass needed to be surgically removed because of the high embolic potential of the mass. Pathologic diagnosis of the mass noted it to be composed of thrombus. In this patient, we theorize that there was an infectious process to cause a bacteremia that may have damaged the intima of the aorta and caused seeding with thrombus. PMID- 19285938 TI - From promiscuity to precision: protein phosphatases get a makeover. AB - The control of biological events requires strict regulation using complex protein phosphorylation and dephosphorylation strategies. The bulk of serine-threonine dephosphorylations are catalyzed by a handful of phosphatase catalytic subunits, giving rise to the misconception that these phosphatases are promiscuous and unregulated enzymes in vivo. The reality is much more nuanced: PP1 and PP2A, the most abundant serine-threonine phosphatases, are, in fact, families of hundreds of protein serine/threonine phosphatases, assembled from a few catalytic subunits in combination with a highly diverse array of regulators. As recent publications illustrate, these regulatory subunits confer specificity, selectivity, localization, and regulation on these important enzymes. PMID- 19285939 TI - Single-stranded DNA orchestrates an ATM-to-ATR switch at DNA breaks. AB - ATM and ATR are two master checkpoint kinases activated by double-stranded DNA breaks (DSBs). ATM is critical for the initial response and the subsequent ATR activation. Here we show that ATR activation is coupled with loss of ATM activation, an unexpected ATM-to-ATR switch during the biphasic DSB response. ATM is activated by DSBs with blunt ends or short single-stranded overhangs (SSOs). Surprisingly, the activation of ATM in the presence of SSOs, like that of ATR, relies on single- and double-stranded DNA junctions. In a length-dependent manner, SSOs attenuate ATM activation and potentiate ATR activation through a swap of DNA-damage sensors. Progressive resection of DSBs directly promotes the ATM-to-ATR switch in vitro. In cells, the ATM-to-ATR switch is driven by both ATM and the nucleases participating in DSB resection. Thus, single-stranded DNA orchestrates ATM and ATR to function in an orderly and reciprocal manner in two distinct phases of DSB response. PMID- 19285940 TI - Sumoylation of RecQ helicase controls the fate of dysfunctional telomeres. AB - Genome stability depends upon the RecQ helicases, which are conserved from bacteria to man, but little is known about how their myriad activities are regulated. Fission yeast lacking the telomere protein Taz1 (mammalian TRF1/TRF2 ortholog) lose many hallmarks of telomeres, including accurate replication and local protection from DNA repair reactions. Here we show that the RecQ homolog, Rqh1, is sumoylated. Surprisingly, Rqh1 acts on taz1Delta telomeres in a deleterious way, promoting telomere breakage and entanglement. Mutation of Rqh1 sumoylation sites rescues taz1Delta cells from these hazards without dramatically affecting nontelomeric Rqh1 functions. The prominence of Rqh1 in the etiology of several different telomere defects supports the idea that they originate from a common underlying lesion--aberrant processing of the stalled telomeric replication forks that accumulate in the absence of Taz1. Our work underscores the principle that RecQ helicases are "double-edged swords" whose activity, while necessary for maintaining genome-wide stability, must be vigilantly controlled. PMID- 19285941 TI - Protection from isopeptidase-mediated deconjugation regulates paralog-selective sumoylation of RanGAP1. AB - Vertebrates express three small ubiquitin-related modifiers (SUMO-1, SUMO-2, and SUMO-3) that are conjugated in part to unique subsets of proteins and, thereby, regulate distinct cellular processes. Mechanisms regulating paralog-selective sumoylation, however, remain poorly understood. Despite being equally well modified by SUMO-1 and SUMO-2 in vitro, RanGAP1 is selectively modified by SUMO-1 in vivo. We have found that this paralog-selective modification is determined at the level of deconjugation by isopeptidases. Our findings indicate that, relative to SUMO-2-modified RanGAP1, SUMO-1-modified RanGAP1 forms a more stable, higher affinity complex with the nucleoporin Nup358/RanBP2 that preferentially protects it from isopeptidases. By swapping residues in SUMO-1 and SUMO-2 responsible for Nup358/RanBP2 binding, or by manipulating isopeptidase expression levels, paralog selective modification of RanGAP1 could be affected both in vitro and in vivo. Thus, protection from isopeptidases, through interactions with SUMO-binding proteins, represents an important mechanism defining paralog-selective sumoylation. PMID- 19285942 TI - Functionally distinct isoforms of Cik1 are differentially regulated by APC/C mediated proteolysis. AB - Cik1, in association with the kinesin Kar3, controls both the mitotic spindle and nuclear fusion during mating. Here, we show that there are two Cik1 isoforms, and that the mitotic form includes an N-terminal domain required for ubiquitination by the Anaphase-Promoting Complex/Cyclosome (APC/C). During vegetative growth, Cik1 is expressed during mitosis and regulates the mitotic spindle, allowing for accurate chromosome segregation. After mitosis, APC/C(Cdh1) targets Cik1 for ubiquitin-mediated proteolysis. Upon exposure to the mating pheromone alpha factor, a smaller APC/C-resistant Cik1 isoform is expressed from an alternate transcriptional start site. This shorter Cik1 isoform is stable and cannot be ubiquitinated by APC/C(Cdh1). Moreover, the two Cik1 isoforms are functionally distinct. Cells that express only the long isoform have defects in nuclear fusion, whereas cells expressing only the short isoform have an increased rate of chromosome loss. These results demonstrate a coupling of transcriptional regulation and APC/C-mediated proteolysis. PMID- 19285943 TI - ESRP1 and ESRP2 are epithelial cell-type-specific regulators of FGFR2 splicing. AB - Cell-type-specific expression of epithelial and mesenchymal isoforms of Fibroblast Growth Factor Receptor 2 (FGFR2) is achieved through tight regulation of mutually exclusive exons IIIb and IIIc, respectively. Using an application of cell-based cDNA expression screening, we identified two paralogous epithelial cell-type-specific RNA-binding proteins that are essential regulators of FGFR2 splicing. Ectopic expression of either protein in cells that express FGFR2-IIIc caused a switch in endogenous FGFR2 splicing to the epithelial isoform. Conversely, knockdown of both factors in cells that express FGFR2-IIIb by RNA interference caused a switch from the epithelial to mesenchymal isoform. These factors also regulate splicing of CD44, p120-Catenin (CTNND1), and hMena (ENAH), three transcripts that undergo changes in splicing during the epithelial-to mesenchymal transition (EMT). These studies suggest that Epithelial Splicing Regulatory Proteins 1 and 2 (ESRP1 and ESRP2) are coordinators of an epithelial cell-type-specific splicing program. PMID- 19285944 TI - A conserved docking surface on calcineurin mediates interaction with substrates and immunosuppressants. AB - The phosphatase calcineurin, a target of the immunosuppressants cyclosporin A and FK506, dephosphorylates NFAT transcription factors to promote immune activation and development of the vascular and nervous systems. NFAT interacts with calcineurin through distinct binding motifs: the PxIxIT and LxVP sites. Although many calcineurin substrates contain PxIxIT motifs, the generality of LxVP mediated interactions is unclear. We define critical residues in the LxVP motif, and we demonstrate its binding to a hydrophobic pocket at the interface of the two calcineurin subunits. Mutations in this region disrupt binding of mammalian calcineurin to NFATC1 and the interaction of yeast calcineurin with substrates including Rcn1, which contains an LxVP motif. These mutations also interfere with calcineurin-immunosuppressant binding, and an LxVP-based peptide competes with immunosuppressant-immunophilin complexes for binding to calcineurin. These studies suggest that LxVP-type sites are a common feature of calcineurin substrates, and that immunosuppressant-immunophilin complexes inhibit calcineurin by interfering with this mode of substrate recognition. PMID- 19285945 TI - PINK1-associated Parkinson's disease is caused by neuronal vulnerability to calcium-induced cell death. AB - Mutations in PINK1 cause autosomal recessive Parkinson's disease. PINK1 is a mitochondrial kinase of unknown function. We investigated calcium homeostasis and mitochondrial function in PINK1-deficient mammalian neurons. We demonstrate physiologically that PINK1 regulates calcium efflux from the mitochondria via the mitochondrial Na(+)/Ca(2+) exchanger. PINK1 deficiency causes mitochondrial accumulation of calcium, resulting in mitochondrial calcium overload. We show that calcium overload stimulates reactive oxygen species (ROS) production via NADPH oxidase. ROS production inhibits the glucose transporter, reducing substrate delivery and causing impaired respiration. We demonstrate that impaired respiration may be restored by provision of mitochondrial complex I and II substrates. Taken together, reduced mitochondrial calcium capacity and increased ROS lower the threshold of opening of the mitochondrial permeability transition pore (mPTP) such that physiological calcium stimuli become sufficient to induce mPTP opening in PINK1-deficient cells. Our findings propose a mechanism by which PINK1 dysfunction renders neurons vulnerable to cell death. PMID- 19285946 TI - SRC-induced disassembly of adherens junctions requires localized phosphorylation and degradation of the rac activator tiam1. AB - The Rac activator Tiam1 is required for adherens junction (AJ) maintenance, and its depletion results in AJ disassembly. Conversely, the oncoprotein Src potently induces AJ disassembly and epithelial-mesenchymal transition (EMT). Here, we show that Tiam1 is phosphorylated on Y384 by Src. This occurs predominantly at AJs, is required for Src-induced AJ disassembly and cell migration, and creates a docking site on Tiam1 for Grb2. We find that Tiam1 is associated with ERK. Following recruitment of the Grb2-Sos1 complex, ERK becomes activated and triggers the localized degradation of Tiam1 at AJs, likely involving calpain proteases. Furthermore, we demonstrate that, in human tumors, Y384 phosphorylation positively correlates with Src activity, and total Tiam1 levels are inversely correlated. Thus, our data implicate Tiam1 phosphorylation and consequent degradation in Src-mediated EMT and resultant cell motility and establish a paradigm for regulating local concentrations of Rho-GEFs. PMID- 19285947 TI - Resampling and editing of mischarged tRNA prior to translation elongation. AB - Faithful translation of the genetic code depends on the GTPase EF-Tu delivering correctly charged aminoacyl-tRNAs to the ribosome for pairing with cognate codons. The accurate coupling of cognate amino acids and tRNAs by the aminoacyl tRNA synthetases is achieved through a combination of substrate specificity and product editing. Once released by aminoacyl-tRNA synthetases, both cognate and near-cognate aminoacyl-tRNAs were considered to be committed to ribosomal protein synthesis through their association with EF-Tu. Here we show instead that aminoacyl-tRNAs in ternary complex with EF-Tu*GTP can readily dissociate and rebind to aminoacyl-tRNA synthetases. For mischarged species, this allows resampling by the product editing pathway, leading to a reduction in the overall error rate of aminoacyl-tRNA synthesis. Resampling of mischarged tRNAs was shown to increase the accuracy of translation over ten fold during in vitro protein synthesis, supporting the presence of an additional quality control step prior to translation elongation. PMID- 19285948 TI - Structural basis for the mutually exclusive anchoring of P body components EDC3 and Tral to the DEAD box protein DDX6/Me31B. AB - The DEAD box helicase DDX6/Me31B functions in translational repression and mRNA decapping. How particular RNA helicases are recruited specifically to distinct functional complexes is poorly understood. We present the crystal structure of the DDX6 C-terminal RecA-like domain bound to a highly conserved FDF sequence motif in the decapping activator EDC3. The FDF peptide adopts an alpha-helical conformation upon binding to DDX6, occupying a shallow groove opposite to the DDX6 surface involved in RNA binding and ATP hydrolysis. Mutagenesis of Me31B shows the relevance of the FDF interaction surface both for Me31B's accumulation in P bodies and for its ability to repress the expression of bound mRNAs. The translational repressor Tral contains a similar FDF motif. Together with mutational and competition studies, the structure reveals why the interactions of Me31B with EDC3 and Tral are mutually exclusive and how the respective decapping and translational repressor complexes might hook onto an mRNA substrate. PMID- 19285950 TI - Knockdown of the PsbP protein does not prevent assembly of the dimeric PSII core complex but impairs accumulation of photosystem II supercomplexes in tobacco. AB - The PsbP protein is an extrinsic subunit of photosystem II (PSII) specifically found in land plants and green algae. Using PsbP-RNAi tobacco, we have investigated effects of PsbP knockdown on protein supercomplex organization within the thylakoid membranes and photosynthetic properties of PSII. In PsbP RNAi leaves, PSII dimers binding the extrinsic PsbO protein could be formed, while the light-harvesting complex II (LHCII)-PSII supercomplexes were severely decreased. Furthermore, LHCII and major PSII subunits were significantly dephosphorylated. Electron microscopic analysis showed that thylakoid grana stacking in PsbP-RNAi chloroplast was largely disordered and appeared similar to the stromally-exposed or marginal regions of wild-type thylakoids. Knockdown of PsbP modified both the donor and acceptor sides of PSII; In addition to the lower water-splitting activity, the primary quinone Q(A) in PSII was significantly reduced even when the photosystem I reaction center (P700) was noticeably oxidized, and thermoluminescence studies suggested the stabilization of the charged pair, S(2)/Q(A)(-). These data indicate that assembly and/or maintenance of the functional MnCa cluster is perturbed in absence of PsbP, which impairs accumulation of final active forms of PSII supercomplexes. PMID- 19285949 TI - Physiological and pharmacological features of the novel gasotransmitter: hydrogen sulfide. AB - Hydrogen sulfide (H(2)S) has been known for hundreds of years because of its poisoning effect. Once the basal bio-production became evident its pathophysiological role started to be investigated in depth. H(2)S is a gas that can be formed by the action of two enzymes, cystathionine gamma-lyase and cystathionine beta-synthase, both involved in the metabolism of cysteine. It has several features in common with the other two well known "gasotransmitters" (nitric oxide and carbon monoxide) in the biological systems. These three gasses share some biological targets; however, they also have dissimilarities. For instance, the three gases target heme-proteins and open K(ATP) channels; H(2)S as NO is an antioxidant, but in contrast to the latter molecule, H(2)S does not directly form radicals. In the last years H(2)S has been implicated in several physiological and pathophysiological processes such as long term synaptic potentiation, vasorelaxation, pro- and anti-inflammatory conditions, cardiac inotropism regulation, cardioprotection, and several other physiological mechanisms. We will focus on the biological role of H(2)S as a molecule able to trigger cell signaling. Our attention will be particularly devoted on the effects in cardiovascular system and in cardioprotection. We will also provide available information on H(2)S-donating drugs which have so far been tested in order to conjugate the beneficial effect of H(2)S with other pharmaceutical properties. PMID- 19285951 TI - High affinity interaction between histidine-rich glycoprotein and the cell surface type ATP synthase on T-cells. AB - Histidine-rich glycoprotein (HRG) is a plasma protein implicated in the innate immune system. In recent studies, we showed that either HRG, or the Arg23-Lys66 glycopeptide derived from HRG, in concert with concanavalin A (Con A), promotes a morphological change and adhesion of the human leukemic T-cell line MOLT-4 to culture dishes, and that cell surface glycosaminoglycan or Fcgamma receptors do not participate in this cellular event. In the present study, we identified the alpha-subunit of ATP synthase as one of the HRG-binding proteins on the surface of T-cells by HRG-derived glycopeptide affinity chromatography and by a peptide mass finger printing method. HRG specifically interacted with mitochondrial ATP synthase with a dissociation constant of 66 nM. The presence of alpha- and beta subunits of ATP synthase on the plasma membrane of MOLT-4 cell was demonstrated by immunofluorescent staining and FACS analysis. The HRG/Con A-induced morphological changes of MOLT-4 cells were specifically inhibited by a monoclonal antibody against the beta-subunit of ATP synthase. These results strongly suggest that the cell surface ATP synthase functions as a binding protein for HRG on MOLT 4 cells, which is required for the morphological changes observed in MOLT-4 cells following treatment with HRG/Con A. PMID- 19285952 TI - Topoisomerase IIalpha inhibition following DNA transfection greatly enhances random integration in a human pre-B lymphocyte cell line. AB - DNA transfection can be too inefficient to establish a desired number of stable transfectants, particularly in lymphocytes; however, this could be circumvented by increasing the absolute frequency of random integration. In this paper, we show that treating cells with topoisomerase II inhibitor following electroporation greatly (approximately 10- to 20-fold) enhances random integration of input DNA in a human pre-B lymphocyte cell line, Nalm-6. With the use of various kinds of topoisomerase II-targeting agents, we also present evidence that topoisomerase IIalpha inhibition is critical for the enhancement of random integration, while the contribution of topoisomerase IIbeta may be negligible. As topoisomerase IIalpha is highly expressed in vigorously growing cells, our results show that topoisomerase IIalpha inhibition provides a promising way of enhancing random integration in virtually all cultured cell lines. PMID- 19285953 TI - Respiratory arsenate reductase as a bidirectional enzyme. AB - The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe-S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism. PMID- 19285954 TI - Tumorigenic properties of alternative osteopontin isoforms in mesothelioma. AB - Osteopontin (SPP1) is an inflammatory cytokine that we previously characterized as a diagnostic marker in patients with asbestos-induced malignant mesothelioma (MM). While SPP1 shows both pro- and anti-tumorigenic biological effects, little is known about the molecular basis of these activities. In this study, we demonstrate that while healthy pleura possesses all three differentially spliced SPP1 isoforms (A-C), in clinical MM specimens isoform A is markedly up-regulated and predominant. To provide a clue to possible functions of the SPP1 isoforms we next performed their functional evaluation via transient expression in MM cell lines. As a result, we report that isoforms A-C demonstrate different activities in cell proliferation, wound closure, and invasion assays. These findings suggest different functions for SPP1 isoforms and underline pro-tumorigenic properties of isoforms A and B. PMID- 19285955 TI - IEX-1-induced cell death requires BIM and is modulated by MCL-1. AB - MCL-1 (myeloid cell leukemia-1) is a distinguished and pivotal member of the pro survival BCL-2 family of proteins, and we isolated IEX-1 (immediate early response gene X-1) as a MCL-1-interacting protein using the yeast two-hybrid system and confirmed their endogenous association in human cells. The underlying mechanisms by which IEX-1 affects cell survival and death are largely unknown. Ectopic expression of IEX-1-induced caspase-dependent apoptosis in 293T cells, and the response was significantly modulated by changes in the MCL-1 expression level in cells. Forced expression of IEX-1 was unable to induce cell death or to perturb mitochondrial membrane potential in BIM-depleted cells. Additionally, knockouts of NOXA or PUMA did not affect the activities of IEX-1, indicating that the pro-death action of IEX-1 specifically requires BIM. Our findings provide insight into a new regulatory circuit that controls cell death and survival by the coordinated action of MCL-1, IEX-1, and BIM. PMID- 19285956 TI - The steady-state expression of connexin43 is maintained by the PI3K/Akt in osteoblasts. AB - The gap junction protein connexin43 (Cx43) plays an important role in bone development and its homeostasis. Therefore, it is important that the Cx43 expression is kept at a high-level in bone tissues under normal conditions. To investigate the mechanism to keep Cx43 expression level, the effects of protein kinase inhibitors on the basal expression of Cx43 were examined. It was found that the specific PI3K inhibitor LY294002 significantly decreased the steady state expression levels of Cx43 mRNA and protein in osteoblastic cell line, MC3T3 E1 cells. Furthermore, dominant-negative Akt expression reduced both Cx43 expression and gap junction activity. These results suggest an important role of PI3K/Akt in the regulation of basal Cx43 expression. A promoter assay for Cx43 and an actinomycin D chase experiment revealed that PI3K/Akt modulated Cx43 expression post-transcriptionally via mRNA stability. The present findings could provide new insights into the molecular understanding of Cx43 expression. PMID- 19285957 TI - Upregulation of immunity-related GTPase (IRG) proteins by TNF-alpha in murine astrocytes. AB - We examined the effect of tumor necrosis factor-alpha (TNF-alpha) on murine primary astrocytes. Proteomic analysis demonstrated that four new spots in the TNF-alpha-treated cells relative to untreated cells. Two of them were identified as Irgb6 and Irgd, members of immunity-related GTPase (IRG) proteins which are the key mediators of interferon-gamma (IFN-gamma)-induced resistance of pathogens in numerous cells. Gene expression analysis using RT-PCR showed that TNF-alpha dose-dependently increased the expression of both proteins. Immunocytochemical analysis showed that TNF-alpha increased the abundance of both proteins. A subcellular localization study demonstrated that TNF-alpha induced the partial colocalization of both proteins with the endoplasmic reticulum (ER) and Golgi apparatus, whereas IFN-gamma did not induce the colocalization of Irgd protein with the ER and Golgi. Combined stimulation with TNF-alpha and IFN-gamma had a synergistic effect on the expression of Irgb6 and an added effect on the expression of Irgd. PMID- 19285958 TI - The MAP1-LC3 conjugation system is involved in lipid droplet formation. AB - Lipid droplets (LDs) are ubiquitous in eukaryotic cells, while excess free fatty acids and glucose in plasma are converted to triacylglycerol (TAG) and stored as LDs. However, the mechanism for the generation and growth of LDs in cells is largely unknown. We show here that the LC3 lipidation system essential for macroautophagy is involved in LD formation. LD formation accompanied by accumulation of TAG induced by starvation was largely suppressed in the hepatocytes that cannot execute autophagy. Under starvation conditions, LDs in addition to autophagosomes were abundantly formed in the cytoplasm of these tissue cells. Moreover, LC3 was localized on the surface of LDs and LC3-II (lipidation form) was fractionated to a perilipin (LD marker)-positive lipid fraction from the starved liver. Taken together, these results indicate that the LC3 conjugation system is critically involved in lipid metabolism via LD formation. PMID- 19285959 TI - FGFR1 forms an FRS2-dependent complex with mTOR to regulate smooth muscle marker gene expression. AB - Vascular smooth muscle cells (VSMCs) switch from a contractile to a synthetic phenotype in human cardiovascular disease such as atherosclerosis and restenosis after angioplasty. VSMCs show reduced expression of contractile proteins and are capable of responding to mitogens by increasing expression of growth factor receptors. Fibroblast growth factor receptor-1 (FGFR1) signaling is one of several signaling pathways involved in this VSMC phenotypic switching. The aim of this study was to examine the signaling pathway downstream of FGFR1 in the regulation of SM marker gene expression. We found that FGFR1 activated Akt/mTOR pathway and that the mTOR inhibitor rapamycin partially reversed FGFR1-mediated downregulation of SM marker gene expression. Furthermore, we showed that mTOR forms a multi-protein complex with FGFR1 in VSMCs. These findings provide novel information for future development of therapeutic strategies for the treatment of human cardiovascular disease. PMID- 19285960 TI - Drosophila melanogaster deoxyribonucleoside kinase activates gemcitabine. AB - Drosophila melanogaster multisubstrate deoxyribonucleoside kinase (Dm-dNK) can additionally sensitize human cancer cell lines towards the anti-cancer drug gemcitabine. We show that this property is based on the Dm-dNK ability to efficiently phosphorylate gemcitabine. The 2.2A resolution structure of Dm-dNK in complex with gemcitabine shows that the residues Tyr70 and Arg105 play a crucial role in the firm positioning of gemcitabine by extra interactions made by the fluoride atoms. This explains why gemcitabine is a good substrate for Dm-dNK. PMID- 19285961 TI - Regulation of DNA repair by parkin. AB - Mutation of parkin is one of the most prevalent causes of autosomal recessive Parkinson's disease (PD). Parkin is an E3 ubiquitin ligase that acts on a variety of substrates, resulting in polyubiquitination and degradation by the proteasome or monoubiquitination and regulation of biological activity. However, the cellular functions of parkin that relate to its pathological involvement in PD are not well understood. Here we show that parkin is essential for optimal repair of DNA damage. Parkin-deficient cells exhibit reduced DNA excision repair that can be restored by transfection of wild-type parkin, but not by transfection of a pathological parkin mutant. Parkin also protects against DNA damage-induced cell death, an activity that is largely lost in the pathological mutant. Moreover, parkin interacts with the proliferating cell nuclear antigen (PCNA), a protein that coordinates DNA excision repair. These results suggest that parkin promotes DNA repair and protects against genotoxicity, and implicate DNA damage as a potential pathogenic mechanism in PD. PMID- 19285962 TI - Capsaicin inhibits the IL-6/STAT3 pathway by depleting intracellular gp130 pools through endoplasmic reticulum stress. AB - Capsaicin has been shown to have anti-carcinogenic effects on various tumor cells through multiple mechanisms. It was recently reported that capsaicin inhibited interleukin-6 (IL-6)-induced activation of signal transducer and activator of transcription 3 (STAT3), an anti-apoptotic transcription factor. Here we demonstrate that capsaicin induced downregulation of the IL-6 receptor gp130 within 2h in glial tumors. The downregulation of gp130 was not caused by enhanced degradation of gp130 or by inhibition of mRNA transcription. The downregulation was attributed to translation inhibition of gp130, which was associated with activation of endoplasmic reticulum (ER) stress. The depletion of the intracellular pool of gp130 by capsaicin and an ER stress inducer led to an immediate loss of the IL-6 response due to the short half-life of membrane localized gp130. These results suggest a novel mechanism for the anti-tumor effect of capsaicin. PMID- 19285963 TI - Identification of casein kinase-1 phosphorylation sites on TDP-43. AB - TAR DNA-binding protein of 43 kDa (TDP-43) is deposited as hyperphosphorylated cytoplasmic and intranuclear inclusions in brains of patients with frontotemporal lobar degeneration with ubiquitinated inclusions and amyotrophic lateral sclerosis. In this study, we identified 29 phosphorylation sites on recombinant TDP-43 that are phosphorylated by casein kinase-1 (CK1). Interestingly, 18 of them were located in the C-terminal glycine-rich region of TDP-43. Our results indicate that CK1-mediated phosphorylation may play a role in the pathogenesis of these diseases. PMID- 19285964 TI - Efficient inhibition of fibroblast proliferation and collagen expression by ERK2 siRNAs. AB - Transforming growth factor-beta1 and fibroblast growth factor-2 play very important roles in fibroblast proliferation and collagen expression. These processes lead to the formation of joint adhesions through the SMAD and MAPK pathways, in which ERK2 is supposed to be crucial. Based on these assumptions, lentivirus (LV)-mediated small interfering RNAs (siRNAs) targeting ERK2 were used to suppress the proliferation and collagen expression of rat joint adhesion tissue fibroblasts (RJATFs). Among four siRNAs examined, siRNA1 caused an 84% reduction in ERK2 expression (p<0.01) and was selected as the most efficient siRNA for use in this study. In subsequent experiments, significant downregulation of types I and III collagen were observed by quantitative RT-PCR and Western blot analyses. MTT assays and flow cytometry revealed marked inhibition of RJATF proliferation, but no apoptosis. In conclusion, LV-mediated ERK2 siRNAs may represent novel therapies or drug targets for preventing joint adhesion formation. PMID- 19285965 TI - Frontal eye field stimulation modulates the balance of salience between target and distractors. AB - Natural scenes generally include several possible objects that can be the target for a shift of gaze and attention. The oculomotor system may select a single target by boosting neural activation representing the target, and also by inhibiting neural activity associated with competing alternatives (distractors). We examine the role of the frontal eye field (FEF) in these processes through the effects of single-pulse transcranial magnetic stimulation (TMS) on the distractor related modulation of saccade trajectories. Participants made voluntary saccades to peripheral locations specified by a central arrow-cue. On some trials, visual distractors appeared remote from the target location. The competing distractor produced a deviation of saccade trajectory, away from the distractor location. Single-pulse TMS stimulation of the right frontal eye field increased this distractor-related deviation compared that observed when stimulation was applied to a control site (vertex). The increase in distractor-related deviation of trajectory, following FEF stimulation, was observed for saccades made in both the left and right visual fields and could not be attributed to an effect of TMS on saccade latency. The enhanced distractor-related deviation following FEF stimulation could reflect increased inhibition of the competing distractor, or reduced salience of the endogenous saccade goal. The results are interpreted in light of neurophysiological evidence that the human FEF is involved in the dynamic interaction between competing stimuli for the selection of a candidate target. PMID- 19285966 TI - An ERP investigation of orthographic priming with relative-position and absolute position primes. AB - The present study used event-related potentials to examine the time-course of relative-position and absolute-position orthographic priming. Relative-position priming was examined using primes formed by a concatenated subset of the target word's letters (e.g., cllet/COLLECT vs. dlema/COLLECT), and absolute-position priming was investigated using hyphenated versions of these primes (c-lle t/COLLECT vs. d-lem-a/COLLECT). Both manipulations modulated the ERP waveform starting at around 100 ms post-target onset and extending into the N400 component. The first clear manifestation of priming was found in the N250 component, where hyphenated primes were found to have an earlier, more robust and more widely distributed effect than the concatenated primes. On the other hand, both prime types had similar effects on N400 amplitude. These results provide important information about the time-course of activation of location-specific and location-invariant (word-centered) orthographic representations during visual word recognition. PMID- 19285967 TI - Mechanisms and genes in human strial presbycusis from animal models. AB - Schuknecht proposed a discrete form of presbycusis in which hearing loss results principally from degeneration of cochlear stria vascularis and decline of the endocochlear potential (EP). This form was asserted to be genetically linked, and to arise independently from age-related pathology of either the organ of Corti or cochlear neurons. Although extensive strial degeneration in humans coincides with hearing loss, EPs have never been measured in humans, and age-related EP reduction has never been verified. No human genes that promote strial presbycusis have been identified, nor is its pathophysiology well understood. Effective application of animal models to this issue requires models demonstrating EP decline, and preferably, genetically distinct strains that vary in patterns of EP decline and its cellular correlates. Until recently, only two models, Mongolian gerbils and Tyrp1(B-lt) mice, were known to undergo age-associated EP reduction. Detailed studies of seven inbred mouse strains have now revealed three strains (C57BL/6J, B6.CAST-Cdh23(CAST), CBA/J) showing essentially no EP decline with age, and four strains ranging from modest to severe EP reduction (C57BL/6-Tyr(c 2J), BALB/cJ, CBA/CaJ, NOD.NON-H2(nbl)/LtJ). Collectively, animal models support five basic principles regarding a strial form of presbycusis: 1) Progressive EP decline from initially normal levels as a defining characteristic; 2) Non universality, not all age-associated hearing loss involves EP decline; 3) A clear genetic basis; 4) Modulation by environment or stochastic events; and 5) Independent strial, organ of Corti, and neural pathology. Shared features between human strial presbycusis, gerbils, and BALB/cJ and C57BL/6-Tyr(c-2J) mice further suggest this condition frequently begins with strial marginal cell dysfunction and loss. By contrast, NOD.NON-H2(nbl) mice may model a sequence more closely associated with strial microvascular disease. Additional studies of these and other inbred mouse and rat models should reveal candidate processes and genes that promote EP decline in humans. PMID- 19285968 TI - The impact of hypoxia and low glucose on the release of acetylcholine and ATP from the incubated cat carotid body. AB - The carotid body (CB) is a polymodal sensor which increases its neural output to the nucleus tractus solitarii with a subsequent activation of several reflex cardiopulmonary responses. Current reports identify acetylcholine (ACh) and adenosine triphosphate (ATP) as two essential excitatory neurotransmitters in the cat and rat CBs. This study explored the impact of hypoxia, low glucose, and the two together on the release of both ACh and ATP from two incubated cat CBs. The CBs were prepared with standard procedures in accordance with the policies and regulations of the Institutional Animal Care and Use Committee. When normalized to their controls, a significant increase of ACh in the incubation medium was measured in response to hypoxia, low glucose, and the combined stimuli. When normalized to their controls, a significant increase in ATP in the incubation medium was measured in response to hypoxia and to the combined stimuli. Low glucose generated an increase in ATP which was not statistically significant (P>0.05). Second, normalizing the initial 3-4 or 2-3 min Time Segment of the challenge Stage to the final 3-4 or 2-3 min Time Segment of the control Stage for both ACh and ATP generated significant increases in response to hypoxia, low glucose (ACh only), and the combined stimuli. The data suggested the possibility that in the cat the increased CB neural output in response to low glucose might be due primarily to ACh. PMID- 19285969 TI - A splice variant of the TATA-box binding protein encoding the polyglutamine containing N-terminal domain that accumulates in Alzheimer's disease. AB - Previously we have reported the accumulation of an N-terminal fragment of the TATA-box binding protein (TBP) in Alzheimer's disease brain tissue and here we report the identification of a naturally occurring TBP splice variant as a likely mechanism for its production. The splice variant described here encodes the polyglutamine-containing N-terminal domain of this key transcription factor. We demonstrate the expression of the splice variant mRNA in a variety of human tissues and that the resulting protein forms inclusions in cell culture transfection studies. The unusual properties of the variant protein suggest that it may be functionally relevant in late onset neurodegenerative diseases. PMID- 19285970 TI - ERP correlates of the incidental retrieval of emotional information: effects of study-test delay. AB - Prior studies indicate that, in tests of recognition memory, ERPs elicited by correctly recognized test items differ according to whether the items were encoded in an emotionally arousing or an emotionally neutral study context. These prior studies employed only a relatively brief (ca. 10 min) retention interval, however. The present study contrasted the ERP correlates of incidental emotional retrieval as a function of study-test delay. Pictures of emotionally neutral objects were encoded in association with either emotionally negative or emotionally neutral scenes. In a repeated measures design (N=19), half of the objects were subjected to a recognition memory test 10 min after completion of the study phase, whereas the remainder were tested 24 h later. After the short delay, ERPs elicited by objects paired with emotional vs. neutral backgrounds differed from around 200 ms post-stimulus, the objects paired with the emotional scenes eliciting the more positive-going waveforms. After 24 h, differences between the ERPs elicited by the two classes of object were still apparent from around 200 ms post-stimulus. Strikingly, these effects differed from those obtained 10 min after study in both their polarity and scalp distribution. The early onset of these ERP effects suggests that they may reflect a form of memory independent of the conscious recollection of the associated study contexts. The qualitative differences in the effects at the two retention intervals raise the possibility that the encoded objects were subjected to consolidation processes that differed according to the emotional attributes of their study contexts. PMID- 19285971 TI - Theta and alpha oscillations linked to risk identifications. AB - Our recent functional MRI and event-related potential studies suggest that neural mechanisms underlying identifications of environmental and personal risks are characterized by distinct neural structures and time courses. The current work further investigated the role of non-phase locked activity in dissociation of the neural processes of environmental and personal risks. We recorded electroencephalogram (EEG) from healthy adults when they identified risky and safe environmental or personal events depicted in words or phrases. Event-related synchronization (ERS) and desynchronization (ERD) in association with risky and safe environmental and personal events were calculated using Morlet's wavelet analysis. We found that, relative to the identification of safe environmental events, the identification of risky environmental events induced increased theta band power at 260-380 ms over the frontal cortex and increased alpha band power after 700 ms with a wide scalp distribution. Moreover, theta band powers associated with the identification of risky environmental events correlated positively with subjective ratings of the risk degree of environmental events. In contrast, the identification of risky personal events was linked to decreased alpha band power at 740-900 ms with a wide scalp distribution compared with the identification of safe personal events. Our findings indicate that theta and alpha band neural oscillations are involved in differentiation between environmental and personal risks. PMID- 19285972 TI - Contribution of the pre-SMA to the production of words and non-speech oral motor gestures, as revealed by repetitive transcranial magnetic stimulation (rTMS). AB - An emerging theoretical perspective, largely based on neuroimaging studies, suggests that the pre-SMA is involved in planning cognitive aspects of motor behavior and language, such as linguistic and non-linguistic response selection. Neuroimaging studies, however, cannot indicate whether a brain region is equally important to all tasks in which it is activated. In the present study, we tested the hypothesis that the pre-SMA is an important component of response selection, using an interference technique. High frequency repetitive TMS (10 Hz) was used to interfere with the functioning of the pre-SMA during tasks requiring selection of words and oral gestures under different selection modes (forced, volitional) and attention levels (high attention, low attention). Results show that TMS applied to the pre-SMA interferes selectively with the volitional selection condition, resulting in longer RTs. The low- and high-attention forced selection conditions were unaffected by TMS, demonstrating that the pre-SMA is sensitive to selection mode but not attentional demands. TMS similarly affected the volitional selection of words and oral gestures, reflecting the response-independent nature of the pre-SMA contribution to response selection. The implications of these results are discussed. PMID- 19285973 TI - Siglec-1 on monocytes is a potential risk marker for monitoring disease severity in coronary artery disease. AB - OBJECTIVES: Siglec-1 has long been considered as an important biomarker of the activation of monocyte/macrophage and a type I interferon-specific imprint, but its role in atherosclerosis has not been elucidated. METHODS: We examined the expression of Siglec-1 by flow cytometry and RT-PCR in 83 CAD patients and 38 healthy controls. In addition, the levels of serum lipids, Gensini score, hs-CRP and homocysteine were determined. RESULTS: The transcriptional and protein levels of Siglec-1 on monocytes in CAD patients were significantly increased compared with healthy controls [3.17 versus 1.0, P<0.01; (11.5+/-3.9)% versus (1.8+/ 2.0)%, P<0.01], but the increased Siglec-1 had no correlation with the level of native serum lipids. Interestingly, the expression of Siglec-1 was positively correlated with Gensini score (r=0.338, P=0.015), hs-CRP (r=0.316, P=0.016) and homocysteine level (r=0.224, P=0.042). CONCLUSION: Siglec-1 may be considered as a potential non-invasive indicator for monitoring disease severity and a biomarker for predicting the relative risk of cardiovascular events. PMID- 19285974 TI - Are girls more insulin-resistant than boys? AB - OBJECTIVE: The objective was to determine whether girls were more insulin resistant than boys. DESIGN AND METHODS: Data from 1009 children (508 males) in 10 elementary schools, between April and September, 2007 were collected. BMI, waist circumference (WC), blood pressure, Tanner stage, lipids, insulin, and glucose were obtained. RESULTS: One hundred and sixty five (16.4%) of the children were obese (>95%ile), and 166 (16.5%) were overweight (85-95%ile). Mean HOMA-IR and insulin were higher among 10.0-13.9-year-old girls than boys. Multiple logistic regression using the 3rd quartile of HOMA-IR as the dependent variable showed that only BMI OR=1.18 (95% CI 1.12-1.24; p<0.001), Tanner OR=1.39 (95% CI 1.12-1.73; p=0.003) and triglycerides 1.005 (95% CI 1.00-1.01; p=0.04) were significantly associated with insulin resistance while sex and HDL-C were not. CONCLUSIONS: This study showed that no significant sex-related differences were found, and HOMA-IR was associated with adiposity and pubertal stage suggesting that the higher values of HOMA-IR in girls than in boys could be due to their earlier pubertal development. PMID- 19285976 TI - 4-Methylumbelliferone inhibits hyaluronan synthesis by depletion of cellular UDP glucuronic acid and downregulation of hyaluronan synthase 2 and 3. AB - Hyaluronan accumulation on cancer cells and their surrounding stroma predicts an unfavourable disease outcome, suggesting that hyaluronan enhances tumor growth and spreading. 4-Methylumbelliferone (4-MU) inhibits hyaluronan synthesis and retards cancer spreading in experimental animals through mechanisms not fully understood. These mechanisms were studied in A2058 melanoma cells, MCF-7 and MDA MB-361 breast, SKOV-3 ovarian and UT-SCC118 squamous carcinoma cells by analysing hyaluronan synthesis, UDP-glucuronic acid (UDP-GlcUA) content, and hyaluronan synthase (HAS) mRNA levels. The maximal inhibition in hyaluronan synthesis ranged 22-80% in the cell lines tested. Active glucuronidation of 4-MU produced large quantities of 4-MU-glucuronide, depleting the cellular UDP-GlcUA pool. The maximal reduction varied between 38 and 95%. 4-MU also downregulated HAS mRNA levels: HAS3 was 84-60% lower in MDA-MB-361, A2058 and SKOV-3 cells. HAS2 was the major isoenzyme in MCF-7 cells and lowered by 81%, similar to 88% in A2058 cells. These data indicate that both HAS substrate and HAS2 and/or HAS3 mRNA are targeted by 4-MU. Despite different target point sensitivities, the reduction of hyaluronan caused by 4-MU was associated with a significant inhibition of cell migration, proliferation and invasion, supporting the importance of hyaluronan synthesis in cancer, and the therapeutic potential of hyaluronan synthesis inhibition. PMID- 19285975 TI - Differential induction of ethanol-metabolizing CYP2E1 and nicotine-metabolizing CYP2B1/2 in rat liver by chronic nicotine treatment and voluntary ethanol intake. AB - Alcohol and nicotine are frequently co-used and co-abused, and use of both drugs alone can affect hepatic drug metabolism. We investigated the influences of chronic nicotine treatment and voluntary ethanol intake on the induction of rat hepatic cytochrome P450 (CYP) enzymes that metabolize ethanol and nicotine. Rats were trained to voluntarily drink ethanol (6% v/v, 1 h) with nicotine pretreatment for 10 days. Another group of rats were treated with the same nicotine doses alone. Hepatic CYP2E1, CYP2B1/2 and CYP2D1 proteins were assessed by immunoblotting. Nicotine pretreatment (0.4, 0.8 and 1.2 mg/kg) increased voluntary ethanol intake on day 10 by 1.8, 2.0, and 1.4 fold respectively compared to saline pretreatment (P<0.01-0.3). CYP2E1 was increased 1.7, 1.8, and 1.4 fold by the three doses of nicotine alone (P<0.02-0.21); CYP2E1 levels were increased by voluntary ethanol intake alone and a further 2.4, 2.2, and 1.8 fold by 0.4, 0.8, and 1.2 mg/kg nicotine respectively versus saline pretreatment (P<0.002-0.06). CYP2B1/2 proteins were not induced by nicotine alone, but were increased by 2.2-2.5 fold by ethanol drinking (P<0.05). CYP2E1 (r=0.67, P<0.001) and CYP2B1/2 levels (r=0.49, P=0.007) correlated with alcohol consumption on day 10. There was no change in CYP2D1. Chronic nicotine increased voluntary ethanol intake thereby enhancing CYP2E1 and CYP2B1/2 levels. Thus CYPs are regulated not only directly by nicotine and ethanol, but also indirectly via an increase in the ethanol consumption in the presence of nicotine pretreatment. Together this may contribute to the co-abuse of these drugs and alter the metabolism of clinical drugs and endogenous substrates. PMID- 19285977 TI - The role of the cytoskeleton in the formation of gap junctions by Connexin 30. AB - Mutations in the genes that encode Connexin 26 (GJB2) and Connexin 30 (GJB6) are the most common known cause of hereditary nonsyndromic sensorineural deafness. Cx26 and Cx30 share a similar protein structure, as well as the same expression distribution pattern in the cochlea. Cx26 has different intracellular trafficking properties compared to those of Cx43 and Cx32, whose trafficking manner is consistent with the classical membrane protein secretory pathway. Until now, however, the trafficking patterns of Cx30 have not been studied. By means of an immunofluorescence staining approach, we found that the targeting of Cx30 to gap junctions in transfected HeLa cells is not affected by brefeldin A, suggesting a Golgi-independent feature, similar to Cx26. Nocodazole had a minimal effect on assembly and distribution of Cx30 gap junctions. Cytochalasin B-induced actin filament depolymerization, however, affected both the pattern and the distribution of Cx30 gap junctions. Co-localization with and/or interaction between Cx30 and microtubules and cortical actin filaments, but not with the tight/adherens junction protein ZO-1, was confirmed by immunofluorescence and/or immunoprecipitation methods. The results suggest that the cytoskeleton, and especially actin filaments, are important components in the processes of assembly, trafficking and stabilization of Cx30 gap junctions. PMID- 19285978 TI - Extracellular calcium promotes the migration of breast cancer cells through the activation of the calcium sensing receptor. AB - Breast cancer is the most frequent form of cancer in women, with the highest incidence of metastasis to the bone. The reason for the preferential destination to the bone is believed to be due to chemoattractant factors released during bone resorption, which act on the cancer cells facilitating their metastasis. One of the factors released during osteolysis that may mediate breast cancer bone localization is Ca2+. Here, we show that extracellular Ca2+ (Ca2+(o)) acting via the calcium-sensing receptor (CaSR), greatly promotes the migration of bone preferring breast cancer cells. In Boyden Chamber and Scratch Wound migration assays, an increase in breast cancer cell migration was observed at 2.5 mM and 5 mM Ca2+(o) compared to basal levels for three of the four breast cancer cell lines tested. However, a significantly greater migratory response was observed for the highly bone metastatic MDA-MB-231 cells, compared to the MCF7 and T47D, which have a lower metastatic potential in vivo. The BT474 cells, which do not metastasize to the bone, did not respond to elevated concentrations of Ca2+(o) in the migration assays. Inhibition of either ERK1/2 MAPK or phospholipase Cbeta (PLCbeta) led to an abolition of the Ca2+(o)-induced migration, implicating these pathways in the migratory response. Knockdown of the CaSR by siRNA resulted in an inhibition of the Ca2+(o)-induced migration, demonstrating the involvement of this receptor in the effect. These results suggest that the activation of the CaSR by elevated Ca2+(o) concentrations, such as those found near resorbing bone, produces an especially strong chemoattractant effect on bone metastatic breast cancer cells toward the Ca2+-rich environment. PMID- 19285979 TI - Effects of mutations on HIV-1 infectivity and neutralization involving the conserved NNNT amino acid sequence in the gp120 V3 loop. AB - The N-glycan g15 within the HIV-1 gp120 V3 loop efficiently blocks antibodies to facilitate viral escape from humoral immune responses. However, we have isolated primary viruses all lacking the N-glycosylation site g15 due to mutations (NNNT>YRNA, HNTV, SIQK), which showed resistance to neutralizing antibodies present in autologous or heterologous HIV-1 positive sera. When introduced into the NL4-3 background, the sequences YRNA, HNTV and SIQK caused an increase of viral infectivity and resistance to neutralization. Thus, despite the lack of g15, primary isolates can escape from neutralization because of specific mutations of the NNNT sequence altering coreceptor usage. PMID- 19285980 TI - Brefeldin A inhibits expression of DNA packaging proteins and nucleocapsid formation of human cytomegalovirus. AB - In this study we used the fungal antibiotic brefeldin A (BFA) to analyze its effect on viral replication. Analysis by electron microscopy demonstrated that no viral particles were observed in cells treated before the onset of viral replication. In the presence of BFA expression of IE2, MCP, pUL104, pUL56 and pUL89 were reduced, while no or slight effect was observed on expression of pp65, pUL44 and pUL57. Strikingly, real time PCR revealed that de novo viral DNA synthesis is reduced but not completely abolished in the presence of BFA. These results indicated that BFA represents a multi-functional compound leading to inhibition of several steps of viral maturation such as expression of viral DNA packaging proteins and capsid formation. PMID- 19285981 TI - Identification of total and differentially expressed excreted-secreted proteins from Trypanosoma congolense strains exhibiting different virulence and pathogenicity. AB - Animal trypanosomosis is a major constraint to livestock productivity in the tropics and has a significant impact on the life of millions of people globally (mainly in Africa, South America and south-east Asia). In Africa, the disease in livestock is caused mainly by Trypanosoma congolense, Trypanosoma vivax, Trypanosoma evansi and Trypanosoma brucei brucei. The extracellular position of trypanosomes in the bloodstream of their host requires consideration of both the parasite and its naturally excreted-secreted factors (secretome) in the course of pathophysiological processes. We therefore developed and standardised a method to produce purified proteomes and secretomes of African trypanosomes. In this study, two strains of T. congolense exhibiting opposite properties of both virulence and pathogenicity were further investigated through their secretome expression and its involvement in host-parasite interactions. We used a combined proteomic approach (one-dimensional SDS-PAGE and two-dimensional differential in-gel electrophoresis coupled to mass spectrometry) to characterise the whole and differentially expressed protein contents of secretomes. The molecular identification of differentially expressed trypanosome molecules and their correlation with either the virulence process or pathogenicity are discussed with regard to their potential as new diagnostic or therapeutic tools against animal trypanosomosis. PMID- 19285982 TI - Controlling metabolism and cell death: at the heart of mitochondrial calcium signalling. AB - Transient increases in intracellular calcium concentration activate and coordinate a wide variety of cellular processes in virtually every cell type. This review describes the main homeostatic mechanisms that control Ca(2+) transients, focusing on the mitochondrial checkpoint. We subsequently extend this paradigm to the cardiomyocyte and to the interplay between cytosol, endoplasmic reticulum and mitochondria that occurs beat-to-beat in excitation-contraction coupling. The mechanisms whereby mitochondria decode fast cytosolic calcium spikes are discussed in the light of the results obtained with recombinant photoproteins targeted to the mitochondrial matrix of contracting cardiomyocytes. Mitochondrial calcium homeostasis is then highlighted as a crucial point of convergence of the environmental signals that mediate cardiac cell death, both by necrosis and by apoptosis. Altogether we point to a role of the mitochondrion as an integrator of calcium signalling and a fundamental decision maker in cardiomyocyte metabolism and survival. PMID- 19285983 TI - Bone marrow mesenchymal stem cells upregulate transient outward potassium currents in postnatal rat ventricular myocytes. AB - Bone marrow mesenchymal stem cell (BMSC) transplantation has been shown to effectively improve cardiac function in experimental animals and patients with myocardial infarction and heart hypertrophy. BMSCs exert potent effects on cardiomyocytes through the inhibition of cardiac apoptosis, the attenuation of cardiac inflammation, etc. However, novel biological actions of BMSCs on cardiomyocytes remain to be explored. The present study was designed to investigate whether BMSCs affect electrophysiological features of neonatal rat ventricular myocytes (NRVMs). BMSCs and NRVMs were indirectly co-cultured at a ratio of 1:10 with a semi-permeable membrane. We found that compared with mono cultured NRVMs, co-cultured NRVMs exhibited an obvious increase of transient outward potassium current (I(to)), accompanied by significant changes in activation, inactivation and recovery of I(to). Meanwhile, K(V)4.2 mRNA which encodes the channel carrying I(to) was more abundant in co-cultured NRVMs than mono-cultured NRVMs. The increases in basic fibroblast growth factor (bFGF) and insulin growth factor-1 (IGF-1) levels were observed in culture medium of BMSCs. bFGF but not IGF-1 upregulated the K(V)4.2 mRNA expression and enhanced I(to) currents. Taken together, we conclude that BMSCs upregulate I(to) of NRVMs, at least partially, by secreting bFGF that in turn upregulates K(V)4.2 expression and alters the kinetics of I(to). PMID- 19285984 TI - Epoxyeicosatrienoic acids limit damage to mitochondrial function following stress in cardiac cells. AB - Epoxyeicosatrienoic acids (EETs) are polyunsaturated fatty acids synthesized from arachidonic acid by CYP2J2 epoxygenase and inactivated by soluble epoxide hydrolase (sEH or Ephx2) to dihydroxyeicosatrienoic acids. Mitochondrial function following ischemic insult is a critical determinant of reperfusion-induced cell death in the myocardium. The objectives of the current study were to investigate the protective role of EETs in mitochondrial function. Mice with the targeted disruption of the Ephx2 gene, cardiomyocyte-specific overexpression of CYP2J2 or perfused with EETs all have improved postischemic LVDP recovery compared to wild type (WT). Perfusion with the mPTP opener, atractyloside, abolished the improved postischemic functional recovery observed in CYP2J2 Tr, sEH null and EET perfused hearts. Electron micrographs demonstrated WT hearts to have increased mitochondrial fragmentation and T-tubule swelling compared to CYP2J2 Tr hearts following 20 min global ischemia and 20 min reperfusion. Direct effects of EETs on mitochondria were assessed in isolated rat cardiomyocytes and H9c2 cells. Laser-induced loss of mitochondrial membrane potential (DeltaPsi(m)) and mPTP opening was significantly reduced in cells treated with 14, 15-EET (1 microM). The EET protective effect was blocked by the putative EET antagonist 14,15 epoxyeicosa-5(Z)-enoic acid (1 muM, 14, 15-EEZE), paxilline (10 microM, BK(Ca) inhibitor) and 5HD (100 microM, K(ATP) inhibitor). Our studies show that EETs can limit mitochondrial dysfunction following cellular stress via a K(+) channel dependent mechanism. PMID- 19285985 TI - Globins synthesize the second messenger bis-(3'-5')-cyclic diguanosine monophosphate in bacteria. AB - Globin-coupled sensors are heme-binding signal transducers in Bacteria and Archaea in which an N-terminal globin controls the activity of a variable C terminal domain. Here, we report that BpeGReg, a globin-coupled diguanylate cyclase from the whooping cough pathogen Bordetella pertussis, synthesizes the second messenger bis-(3'-5')-cyclic diguanosine monophosphate (c-di-GMP) upon oxygen binding. Expression of BpeGReg in Salmonella typhimurium enhances biofilm formation, while knockout of the BpeGReg gene of B. pertussis results in decreased biofilm formation. These results represent the first identification a signal ligand for any diguanylate cyclase and provide definitive experimental evidence that a globin-coupled sensor regulates c-di-GMP synthesis and biofilm formation. We propose that the synthesis of c-di-GMP by globin sensors is a widespread phenomenon in bacteria. PMID- 19285986 TI - Control elements in the neighboring ATPase gene influence spatiotemporal expression of the human agouti-related protein. AB - The agouti-related protein (AgRP) is an orexigenic peptide that plays a significant role in the regulation of energy balance. It is expressed in the hypothalamus, the adrenal glands, and the testis, but sequences determining its spatial and temporal expression have not been identified. Using an elaborate in vitro screening approach, we show here that two adjacent enhancers inside the first intron of the neighboring (1.4 kb downstream) ATPase gene (ATP6V0D1) modulate the human AgRP promoter with profound spatiotemporal variation despite their diminutive sizes (221 and 231 nt). In transgenic mice, the proximal enhancer displayed specificity for the testis, tail, and ears, and the distal one for the testis, front feet, bone, heart, muscle, brain, spinal cord, and tongue, while dietary fat and overnight fasting had differential effects on enhancer activities. AgRP in the testis was localized to pachytene spermatocytes and in the tongue to epithelial cells. Comparative sequence analysis showed that the AgRP-ATP6V0D1 intergenic region is two times longer in humans than in mice and that the two enhancers are conserved in the rhesus monkey genome but not in the mouse genome. These data show that spatiotemporal expression of the human AgRP gene is influenced by diversified primate-specific intronic sequences in its neighboring ATP6V0D1 gene. PMID- 19285987 TI - Affinity maturation of a humanized rat antibody for anti-RAGE therapy: comprehensive mutagenesis reveals a high level of mutational plasticity both inside and outside the complementarity-determining regions. AB - Antibodies that neutralize RAGE (receptor for advanced glycation end products) ligand interactions have potential therapeutic applications in both acute and chronic diseases. We generated XT-M4, a rat anti-RAGE monoclonal antibody that has in vivo efficacy in an acute sepsis model. This antibody was subsequently humanized. To improve the affinity of this antibody for the treatment of chronic indications, we used random and targeted mutagenesis strategies in combination with ribosome and phage-display technologies, respectively, to generate libraries of XT-M4 variants. We identified a panel of single-chain Fv antibody fragments (scFv's) that was improved up to 110-fold in a homogeneous time-resolved fluorescence competition assay against parental XT-M4 immunoglobulin G (IgG). After reformatting to bivalent scFv-Fc fusions and IgGs, we observed similar gains in potency in the same assay. Further analysis of binding kinetics as IgG revealed multiple variants with subnanomolar apparent affinity that was dictated primarily by improvements in the off-rate. All variants also had improved binding to cell surface-expressed human RAGE, and all retained, or had improved, apparent affinity for mouse RAGE. F100bL in V(H) (variable region of the heavy chain) complementarity-determining region 3 (CDR3) was one of a number of key mutations that correlated with affinity improvements and was independently identified by both mutagenesis strategies. Random mutagenesis coupled with ribosome display and high-throughput screening revealed an unexpectedly high level of mutational plasticity across the whole length of the humanized scFv, suggesting greater scope for structural optimization outside of the primary antigen-combining site defined by V(H) CDR3 and V(kappa) CDR3. In summary, our comprehensive mutagenesis approach not only achieved the desired affinity maturation of XT-M4 but also defined multiple mutational hotspots across the antibody sequence, provided an insight into the specificity-determining residues of the antibody paratope, and identified additional sites within the CDR loops where human germ-line amino acids may be introduced without affecting function. PMID- 19285988 TI - Proton-transfer pathways in photosynthetic reaction centers analyzed by profile hidden markov models and network calculations. AB - In the bacterial reaction center (bRC) of Rhodobacter sphaeroides, the key residues of proton transfer to the secondary quinone (Q(B)) are known. Also, several possible proton entry points and proton-transfer pathways have been proposed. However, the mechanism of the proton transfer to Q(B) remains unclear. The proton transfer to Q(B) in the bRC of Blastochloris viridis is less explored. To analyze whether the bRCs of different species use the same key residues for proton transfer to Q(B), we determined the conservation of these residues. We performed a multiple-sequence alignment based on profile hidden Markov models. Residues involved in proton transfer but not located at the protein surface are conserved or are only exchanged to functionally similar amino acids, whereas potential proton entry points are not conserved to the same extent. The analysis of the hydrogen-bond network of the bRC from R. sphaeroides and that from B. viridis showed that a large network connects Q(B) with the cytoplasmic region in both bRCs. For both species, all non-surface key residues are part of the network. However, not all proton entry points proposed for the bRC of R. sphaeroides are included in the network in the bRC of B. viridis. From our analysis, we could identify possible proton entry points. These proton entry points differ between the two bRCs. Together, the results of the conservation analysis and the hydrogen-bond network analysis make it likely that the proton transfer to Q(B) is not mediated by distinct pathways but by a large hydrogen bond network. PMID- 19285989 TI - The first N-terminal amino acids of alpha-synuclein are essential for alpha helical structure formation in vitro and membrane binding in yeast. AB - Alpha-synuclein (alpha-syn), a protein implicated in Parkinson's disease, is structurally diverse. In addition to its random-coil state, alpha-syn can adopt an alpha-helical structure upon lipid membrane binding or a beta-sheet structure upon aggregation. We used yeast biology and in vitro biochemistry to detect how sequence changes alter the structural propensity of alpha-syn. The N-terminus of the protein, which adopts an alpha-helical conformation upon lipid binding, is essential for membrane binding in yeast, and variants that are more prone to forming an alpha-helical structure in vitro are generally more toxic to yeast. beta-Sheet structure and inclusion formation, on the other hand, appear to be protective, possibly by sequestering the protein from the membrane. Surprisingly, sequential deletion of residues 2 through 11 caused a dramatic drop in alpha helical propensity, vesicle binding in vitro, and membrane binding and toxicity in yeast, part of which could be mimicked by mutating aspartic acid at position 2 to alanine. Variants with distinct structural preferences, identified here by a reductionist approach, provide valuable tools for elucidating the nature of toxic forms of alpha-syn in neurons. PMID- 19285990 TI - Targeting of adenovirus serotype 5 pseudotyped with short fiber from serotype 41 to c-erbB2-positive cells using bispecific single-chain diabody. AB - The purpose of the current study was to alter the broad native tropism of human adenovirus for virus targeting to c-erbB2-positive cancer cells. First, we engineered a single-chain antibody (scFv) against the c-erbB2 oncoprotein into minor capsid protein IX (pIX) of adenovirus serotype 5 (Ad5) in a manner commensurate with virion integrity and binding to the soluble extracellular c erbB2 domain. To ablate native viral tropism and facilitate binding of the pIX incorporated scFv to cellular c-erbB2, we replaced the Ad5 fiber with the Ad41 short (41s) fiber devoid of all known cell-binding determinants. The resultant Ad5F41sIX6.5 vector demonstrated increased cell binding and gene transfer as compared to the Ad5F41s control; however, this augmentation of virus infectivity was not c-erbB2 specific. Incorporation of a six-histidine (His(6)) peptide into the C-terminus of the 41s fiber protein resulted in markedly increased Ad5F41s6H infectivity in 293AR cells, which express a membrane-anchored scFv against the C terminal oligohistidine tag, as compared to the Ad5F41s vector and the parental 293 cells. These data suggested that a 41s-fiber-incorporated His(6) tag could serve for attachment of an adapter protein designed to guide Ad5F41s6H infection in a c-erbB2-specific manner. We therefore engineered a bispecific scFv diabody (scDb) combining affinities for both c-erbB2 and the His(6) tag and showed its ability to provide up to 25-fold increase of Ad5F41s6H infectivity in c-erbB2 positive cells. Thus, Ad5 fiber replacement by a His(6)-tagged 41s fiber coupled with virus targeting mediated by an scDb adapter represents a promising strategy to confer Ad5 vector tropism for c-erbB2-positive cancer cells. PMID- 19285991 TI - Functional analysis of yeast bcs1 mutants highlights the role of Bcs1p-specific amino acids in the AAA domain. AB - The mitochondrial protein Bcs1p is conserved from Saccharomyces cerevisiae to humans and its C-terminal region exhibits an AAA (ATPases associated with diverse cellular activities) domain. The absence of the yeast Bcs1p leads to an assembly defect of the iron-sulfur protein (ISP) subunit within the mitochondrial respiratory complex III, whereas human point mutations located all along the protein cause various pathologies. We have performed a structure-function analysis of the yeast Bcs1p by randomly generating a collection of respiratory deficient point mutants. We showed that most mutations are in the C-terminal region of Bcs1p and have localized them on a theoretical three-dimensional model based on the structure of several AAA proteins. The mutations can be grouped into classes according to their respiratory competence and their location on the three dimensional model. We have further characterized five mutants, each substituting an amino acid conserved in yeast and mammalian Bcs1 proteins but not in other AAA proteins. The effects on respiratory complex assembly and Bcs1p accumulation were analyzed. Intragenic and extragenic compensatory mutations able to restore complex III assembly to the mutants affecting the AAA domain were isolated. Our results bring new insights into the role of specific residues in critical regions that are also conserved in the human Bcs1p. We show that (1) residues located at the junction between the Bcs1p-specific and the AAA domains are important for the activity and stability of the protein and (2) the residue F342 is important for interactions with other partners or substrate proteins. PMID- 19285992 TI - Mechanism of ATP-driven PCNA clamp loading by S. cerevisiae RFC. AB - Circular clamps tether polymerases to DNA, serving as essential processivity factors in genome replication, and function in other critical cellular processes as well. Clamp loaders catalyze clamp assembly onto DNA, and the question of how these proteins construct a topological link between a clamp and DNA, especially the mechanism by which ATP is utilized for the task, remains open. Here we describe pre-steady-state analysis of ATP hydrolysis, proliferating cell nuclear antigen (PCNA) clamp opening, and DNA binding by Saccharomyces cerevisiae replication factor C (RFC), and present the first kinetic model of a eukaryotic clamp-loading reaction validated by global data analysis. ATP binding to multiple RFC subunits initiates a slow conformational change in the clamp loader, enabling it to bind and open PCNA and to bind DNA as well. PCNA opening locks RFC into an active state, and the resulting RFC.ATP.PCNA((open)) intermediate is ready for the entry of DNA into the clamp. DNA binding commits RFC to ATP hydrolysis, which is followed by PCNA closure and PCNA.DNA release. This model enables quantitative understanding of the multistep mechanism of a eukaryotic clamp loader and furthermore facilitates comparative analysis of loaders from diverse organisms. PMID- 19285993 TI - Single-stranded DNA-binding protein complex from Helicobacter pylori suggests an ssDNA-binding surface. AB - Single-stranded DNA (ssDNA)-binding protein (SSB) plays an important role in DNA replication, recombination, and repair. SSB consists of an N-terminal ssDNA binding domain with an oligonucleotide/oligosaccharide binding fold and a flexible C-terminal tail involved in protein-protein interactions. SSB from Helicobacter pylori (HpSSB) was isolated, and the ssDNA-binding characteristics of HpSSB were analyzed by fluorescence titration and electrophoretic mobility shift assay. Tryptophan fluorescence quenching was measured as 61%, and the calculated cooperative affinity was 5.4x10(7) M(-1) with an ssDNA-binding length of 25-30 nt. The crystal structure of the C-terminally truncated protein (HpSSBc) in complex with 35-mer ssDNA [HpSSBc-(dT)(35)] was determined at a resolution of 2.3 A. The HpSSBc monomer folds as an oligonucleotide/oligosaccharide binding fold with a Y-shaped conformation. The ssDNA wrapped around the HpSSBc tetramer through a continuous binding path comprising five essential aromatic residues and a positively charged surface formed by numerous basic residues. PMID- 19285994 TI - The rate at which asexual populations cross fitness valleys. AB - Complex traits often involve interactions between different genetic loci. This can lead to sign epistasis, whereby mutations that are individually deleterious or neutral combine to confer a fitness benefit. In order to acquire the beneficial genotype, an asexual population must cross a fitness valley or plateau by first acquiring the deleterious or neutral intermediates. Here, we present a complete, intuitive theoretical description of the valley-crossing process across the full spectrum of possible parameter regimes. We calculate the rate at which a population crosses a fitness valley or plateau of arbitrary width, as a function of the mutation rates, the population size, and the fitnesses of the intermediates. We find that when intermediates are close to neutral, a large population can cross even wide fitness valleys remarkably quickly, so that valley crossing dynamics may be common even when mutations that directly increase fitness are also possible. Thus the evolutionary dynamics of large populations can be sensitive to the structure of an extended region of the fitness landscape the population may not take directly uphill paths in favor of paths across valleys and plateaus that lead eventually to fitter genotypes. In smaller populations, we find that below a threshold size, which depends on the width of the fitness valley and the strength of selection against intermediate genotypes, valley-crossing is much less likely and hence the evolutionary dynamics are less influenced by distant regions of the fitness landscape. PMID- 19285995 TI - Optimization under frequency-dependent selection. AB - We consider a model of frequency-dependent selection, which we refer to as the Wildcard Model. A variety of more specific models, representing quite diverse biological situations, are covered by the Wildcard Model as particular cases. Two very different particular models that are subsumed by the Wildcard Model are the game theoretically motivated two-phenotype model of Lessard [Lessard, S.,1984. Evolutionary dynamics in frequency-dependent two-phenotype models, Theor. Popul. Biol. 25, 210-234], and the model of selection on a continuous trait due to intraspecific competition of Burger [Burger, R., 2005. A multilocus analysis of intraspecific competition and stabilizing selection on a quantitative trait. J. Math. Biol. 50 (4), 355-396] and Schneider [Schneider, K.A., 2006. A multilocus multiallele analysis of frequency-dependent selection induced by intraspecific competition. J. Math. Biol. 52 (4), 483-523]. Both these models have been shown in the past to have a global Lyapunov function (LF) under appropriate genetic assumptions. We show that (i) the Wildcard Model in continuous time for a single multiallelic locus, or for multiple multiallelic loci in linkage equilibrium, has a global LF, of which the Lessard and Burger-Scheneider LF are special cases in spite of their widely different biological interpretations; (ii) the LF of the Wildcard Model can be derived from an LF previously identified for a model of density- and frequency-dependent selection due to Lotka-Volterra competition, with one locus, multiple alleles, multiple species and continuous-time dynamics [Matessi, C., Jayakar, S.D., 1981. Coevolution of species in competition: A theoretical study. Proc. Natl. Acad. Sci. USA, 78 (2, part2), 1081-1084]. We extend the LF with density and frequency dependence to the multilocus case with linkage-equilibrium dynamics. As a possible application of our results, the optimization principle we established can be used as a tool in the study of long term evolution of various models subsumed by the Wildcard Model based on explicit short-term dynamics. PMID- 19285996 TI - Structures of sea anemone toxins. AB - Sea anemones produce a variety of toxic peptides and proteins, including many ion channel blockers and modulators, as well as potent cytolysins. This review describes the structures that have been determined to date for the major classes of peptide and protein toxins. In addition, established and emerging methods for structure determination are summarized and the prospects for modelling newly described toxins are evaluated. In common with most other classes of proteins, toxins display conformational flexibility which may play a role in receptor binding and function. The prospects for obtaining atomic resolution structures of toxins bound to their receptors are also discussed. PMID- 19285997 TI - Two volatiles from the venom gland of the Samsum ant, Pachycondyla sennaarensis. AB - Allergy and anaphylactic reactions after the stings of the Samsum ant Pachycondyla sennaarensis (Mayr, 1862) (Hymenoptera: Formicidae) have been reported from several countries along the Persian Gulf coast, but no analysis has been yet carried out on the ant's venom gland secretions. This study is focused on the identification of volatiles from the venom gland of Pachycondyla sennaarensis using gas chromatography-mass spectrometry, which showed the presence of The main volatile components of the venom gland were phenol-2,4 bis(1,1 dimethylethyl) and trimethyl pyrazine. This is the first record of the occurrence of phenol-2,4-bis(1,1 dimethylethyl) in insects. The venom gland secretions of Pachycondyla species are known contain a variety of volatiles, making the members of this genus distinctive among the ponerine ants. PMID- 19285998 TI - Beta-N-methylaminoalanine (BMAA): metabolism and metabolic effects in model systems and in neural and other tissues of the rat in vitro. AB - The non-protein amino acid, beta-N-methylaminoalanine (BMAA), is neurotoxic and has been implicated in the amyotrophic lateral sclerosis-Parkinsonism-dementia (ALS-PD) complex of Guam. This concept remains controversial, in part because of the lack of a convincing animal model. The neuropharmacology of BMAA is well established, but little is known of its metabolism. This paper reports aspects of the metabolism, and metabolic effects, of BMAA in rat tissues. BMAA changed the distribution of taurine, glycine and serine between rat brain slices and their incubation medium; the glutamate/glutamine cycle between neurones and glia was also compromised. In model experiments BMAA reacted non-enzymatically with pyridoxal-5'-phosphate, releasing methylamine. Rat liver and kidney homogenates, but not brain homogenates, also formed methylamine and 2,3-diaminopropanoic acid when incubated with BMAA. These results provide evidence that several biochemical mechanisms are involved in the neurotoxicity of BMAA. The novel discovery that methylamine is formed from BMAA in rat liver and kidney preparations may be significant since chronic administration of methylamine to rats causes oxidative stress. The extent to which this reaction occurs in different animal species might be a decisive factor in selecting an animal model. PMID- 19285999 TI - Characterization of inflammatory reaction induced by neuwiedase, a P-I metalloproteinase isolated from Bothrops neuwiedi venom. AB - The Snake Venom Metalloproteinases (SVMPs) play a relevant role in the multifactorial inflammatory response induced by Bothrops envenomations. Neuwiedase, an SVMP isolated from Bothrops neuwiedi venom, is devoid of hemorrhagic activity on skin tests, but is able to induce myonecrosis and degrade fibrinogen, fibrin, type I collagen, fibronectin and laminin. In this study, we analyzed the inflammatory reaction induced by neuwiedase in gastrocnemius muscle, with special focus on cytokines release. Our results showed clear evidence of inflammatory infiltrate in the gastrocnemius muscle and an increase of MMP-9, and the cytokines KC, IL-1 beta and IL-6 in the early periods after toxin injection. The cytokine release was also evaluated in inflammatory and muscular cell culture. Both murine peritoneal adherent cells (MPACs) and muscle cells (C2C12) released pro-inflammatory cytokines after stimulus with neuwiedase. MPACs showed increased production of KC, IL-1 beta and IL-6 in the cell culture supernatant while in C2C12, the predominant chemokine expressed was KC. These data reinforce the importance of SVMPs in the inflammatory response caused by envenomation and point out the role of muscle cells in this event by releasing pro-inflammatory mediators able to attract leukocytes to the muscle, thus starting and amplifying the setting of the inflammatory reaction. PMID- 19286000 TI - Cnidocyst structure and the biomechanics of discharge. AB - The cnidocyst is the defining organelle of the cnidarians, used for capture of prey and defense. It consists of a cylindrical capsule, which releases a long tubule upon triggering. Cnidocysts develop inside a giant post-Golgi vesicle by a sequential accumulation of proteins from the Golgi apparatus. Traditionally three types of cnidocysts are distinguished: nematocysts, spirocysts, and ptychocysts. Here we focus on nematocysts, the prototypic cnidocyst and by far most diverse group of cnidocysts in this phylum. The mature nematocyst capsule comprises a collagenous polymer with remarkable biophysical properties, able to withstand an osmotic pressure of 150 bar. Release of the capsule and discharge is probably initiated by classical exocytosis. High-speed studies revealed the kinetics of discharge to be as short as 700 ns, generating an acceleration of 5,400,000 x g and a pressure of 7.7 GPa at the site of impact of the spines onto the prey. Thus nematocysts comprise a powerful molecular spring mechanism releasing energy stored in the wall polymer in the nanosecond time range. During the last few years, genomic, biochemical and structural studies have helped to unravel the molecular composition of the nematocyst supra-structure. Here we summarize these findings and present an integrative view of mechanical and molecular aspects that have shaped the nematocyst during evolution. PMID- 19286001 TI - NeuroBloc/Myobloc: unique features and findings. AB - This review outlines factors that differentiate botulinum toxin serotypes and focuses on the unique features of the commercially available form of BoNT-B (i.e., Myobloc/NeuroBloc). A series of preclinical studies in Cynomolgus monkeys are reviewed. Each of these studies used electrophysiologic measures of changes in the compound muscle action potential (CMAP) following supramaximal nerve stimulation to evaluate the direct effects of the toxin in the injected muscle, as well as the spread of the effects to non-injected muscles. The results of 14 studies were summarized, including several that compared the effects of equivalent doses of BoNT-A and BoNT-B injected into muscles on the opposite side of the same monkey. There is clear evidence that when equivalent doses of BoNT-A and BoNT-B are assessed, there is greater spread to both nearby and remote non injected muscles associated with BoNT-A. Similar studies in the mouse model demonstrated that high, but non-lethal, doses of BoNT-A unilaterally injected into the foot resulted in spread of the effects across the midline to the opposite non-treated foot, while there was no evidence of bilateral effects with equivalent unilateral injections of BoNT-B. Finally, this review summarizes a series of studies in the trapezius and gastrocnemius muscles of monkeys demonstrating that when doses producing equivalent initial effects of BoNT-A and BoNT-B are compared, the duration of effects and the time course of recovery are almost identical across toxins. PMID- 19286002 TI - Visual processing of the bee innately encodes higher-order image statistics when the information is consistent with natural ecology. AB - Determining the statistical relationships of images that facilitate robust visual behaviour is nontrivial. Here we ask if some spatial relationships are more easily learned by the visual brain than others. Visually naive bumblebees were trained to recognise coloured artificial flowers in scenes of equal spatial complexity but differing patterns of stimulus intensity. When flowers of similar intensity were grouped into extended regions across the array (coincident with natural patterns of light), the accuracy of the bees' foraging behaviour was dependent on spatial context, even though this information was redundant to the task. When the same intensity information was organised into a pattern that was less consistent with natural patterns of illumination but of equal order, their behaviour was independent of spatial context and they required double the training time to solve the same conditional task. These observations suggest the brain is biased to more efficiently encode/learn ecologically 'meaningful' image correlations. PMID- 19286003 TI - Body mass index associated with elevated blood pressure in Mexican school-aged adolescents. AB - OBJECTIVE: To evaluate the relationship between blood pressure and body mass index among adolescents. METHODS: A 14-month period cross-sectional study was carried out among 2387 adolescents attending public schools in the state of Morelos, Mexico between 2005 and 2007. We measured blood pressure during four visits, obtaining elevated blood pressure prevalence according to the guidelines for children and adolescents of the United States National Educational Program on High Blood Pressure (at least three visits needed to assess elevated blood pressure on the basis of 95th percentile specific for gender, age and height). We used specific body mass index percentiles for age and gender. We employed multiple linear and Cox proportional hazards models to identify factors related to elevated blood pressure. RESULTS: The overall prevalence of elevated blood pressure was 3.9%. Multiple linear models showed that overweight subjects had systolic and diastolic blood pressures that were 5.1 and 2.5 mmHg higher, respectively, compared with adolescents with a normal body mass index, while obese subjects had 11.3 and 6.2 mmHg higher levels, respectively. Cox proportional hazards models indicated high risk of elevated blood pressure among overweight (RR, 3.6; 95%CI, 1.5-8.5) and obese subjects (RR, 14.2; 95%CI, 7.2 27.75) compared with adolescents with a normal body mass index. CONCLUSION: Our results suggest that a higher body mass index is associated with elevated systolic and diastolic blood pressure levels, indicating the importance of incorporating strategies for ongoing screening and for promoting educational programs on healthy lifestyles to prevent hypertension in adolescents. PMID- 19286005 TI - Analysis of thyreostats: a history of 35 years. AB - Thyreostatic drugs (TS), illegally administrated to livestock for fattening purposes, are banned in the European Union since 1981 (Council Directive 81/602/EC). This paper reviews the trends in the analytical approaches for the determination of TS drugs in biological matrices. After a brief introduction on the different groups of compounds with a thyreostatic action, the most relevant legislation regarding the residue control of these compounds is presented. An overview of the analytical possibilities for the determination of TS in animal matrices, covering sample extraction, purification, separation techniques and detection methods is provided. Additionally, a brief outline of animal experiments is described that illustrates the excretion and distribution profiles of TS residues. Finally, the novel developments in TS analysis are highlighted. Also the possible semi-endogenous status of thiouracil is discussed. PMID- 19286006 TI - Risks of antibiotic residues in milk following intramammary and intramuscular treatments in dairy sheep. AB - Very few drugs on the market are approved for use in lactating ewes. Veterinarians in the European Union are allowed to prescribe drugs in an off label manner but are then obligated to assure that residues do not enter the food chain. In case of mastitis treatment in dairy ewes antibiotic preparations designed and authorized for the bovine mammary gland are usually used. Due to inter-species differences, available bovine data cannot be accurately extrapolated for use in the dairy ewe. The objective of the study was therefore to determine appropriate withdrawal periods for ewe's milk following mastitis treatment with two commercial lactating cow products. For the detection of all components standard agar plate diffusion techniques were used. Regardless of the therapy regime and the product used, residues of antibiotics in milk were detected up to 192h after the last infusion. These results indicate that the required withholding periods for ewe's milk are considerably longer than recommended on the label for bovine milk. PMID- 19286007 TI - Development and validation of a confirmatory method for the determination of sulphonamides in milk by liquid chromatography with diode array detection. AB - A simple and rapid multiresidue method for the determination of seven sulphonamides residues (sulfadiazine, sulfapyridine, sulfamerazine, sulfamethazine, sulfamonomethoxine, sulfadimethoxine and sulfaquinoxaline) in milk samples was developed and validated. The drugs were extracted with a mixture chloroform/acetone and simply cleaned up on a cation exchange solid phase extraction column. The analytes determination was carried out using liquid chromatography with diode array detection (DAD). The procedure has validated as a quantitative confirmatory method according to the European Union (EU) Decision 2002/657/EC. The developed method shows good linearity, specificity, precision (repeatability and intra-laboratory reproducibility), ruggedness and is able to confirm each sulphonamide residue above 30mugkg(-1). Decision limits (CCalpha) around 110mugkg(-1) and recovery above 56% were obtained for all the analytes. The results of the validation process demonstrate that the method is suitable for application, as confirmatory method, in European Union statutory veterinary drug residue surveillance programmes. In addition, a hypothetical situation of sample judgement (compliance or not) in the case in which, at the same time, two different sulphonamides are found, is discussed. PMID- 19286008 TI - Development, validation and implementation of a receptor based bioassay capable of detecting a broad range of beta-agonist drugs in animal feedingstuffs. AB - A bioassay was developed for the detection of a broad range of beta-agonist compounds in animal feeds. A solubilised beta2-adrenoceptor was utilised as the binding protein in the assay. This protein was found to be highly stable when stored at 80 degrees C. The assay was developed and initially validated to determine the sensitivity and relative selectivity against a panel of commonly used beta-agonist compounds. It was also shown that when beta-agonists were present as cocktails in samples a pronounced synergistic effect could be measured. The method was further validated according to EC Decision 2002/657 and proved capable of detecting 250ng clenbuterol equivalents per gram of sample. This is well below the quantities normally associated with beta-agonist medicated feeds. The beta2-adrenoceptor used in the study only failed to bind the compound zilpaterol, raising doubts as to whether this compound is a true beta2-adrenergic drug. PMID- 19286009 TI - Concentrations of danofloxacin 18% solution in plasma, milk and tissues after subcutaneous injection in dairy cows. AB - Danofloxacin is a fluoroquinolone developed for use in veterinary medicine. Its concentrations and pharmacokinetic profile in plasma, milk and tissues of lactating dairy cows were determined, and its milk withdrawal time (WT) calculated. Twenty-one dairy cows received a single subcutaneous administration of 18% mesylate danofloxacin salt (6 mgkg(-1)). Plasma and milk samples were obtained at different times until 48 h. Groups of three animals were sacrificed at different post-administration times and tissue samples (mammary gland, uterus, duodenum, jejunum, ileum, colon and mesenteric lymph nodes) obtained. Danofloxacin concentrations were determined by liquid chromatography with fluorescence detection. The milk WT was calculated by the Time to Safe Concentration method (Software WTM 1.4, EMEA). Danofloxacin was rapidly absorbed and its distribution from plasma to all sampled tissues and milk was extensive. Milk and tissues concentrations were several times above those found in plasma. Plasma area under the curve (AUCp) was 9.69 microghmL(-1) and its elimination half life (T(beta)(1/2)) was 12.53 h. AUC values for the various tissues and milk greatly exceeded AUCp. T(beta)(1/2) from milk and tissues ranged between 4.57 and 21.91 h and the milk withdrawal time was 73.48 h. The reported results support the potential use of danofloxacin in the treatment of mastitis and other infections in milk cows with 3 days of withdrawal. PMID- 19286010 TI - A confirmatory method for the determination of tetracyclines in muscle using high performance liquid chromatography with diode-array detection. AB - Using high-performance liquid chromatography with diode-array detection (HPLC DAD) technique, a confirmatory method for the determination of trace levels of tetracyclines (oxytetracycline, tetracycline, chlortetracycline and doxycycline) and their 4-epimers (4-epioxytetracycline, 4-epitetracycline and 4 epichlortetracycline) in animal tissues (muscle) was developed. The samples are extracted with a mixture of succinic acid 0.1M (pH 4) and methanol after the addition of metacycline as internal standard. The clean-up is carried out by metal chelate affinity chromatography with a following concentration step on an OASIS HLB polymeric reversed phase column. The chromatographic separation of the seven analytes is achieved in 10 min on a short monolithic column (50 mm x 4.6 mm i.d.) using a gradient elution. The method was validated in bovine muscle following the Commission Decision 2002/657/EC criteria: samples spiked at four concentration levels (0.25, 0.5, 1 and 1.5 times the maximum residue limit) were analysed. Method trueness and precision (repeatability and intra-laboratory reproducibility) as well as decision limits (CCalpha) and detection capabilities (CCbeta) are reported. PMID- 19286011 TI - Confirmatory method for the determination of resorcylic acid lactones in urine sample using immunoaffinity cleanup and liquid chromatography-tandem mass spectrometry. AB - The presence of zeranol (alpha-zearalanol) in urine samples due to natural contamination or illegal treatment is under debate within the European Union. The simultaneous determination of zeranol, its epimer taleranol (beta-zearalanol), zearalanone and the structurally related mycotoxin zearalenone with the corresponding alpha- and beta-zearalenol metabolites appears to be critical in deciding whether an illegal use has occurred. The aim of this study is to develop and validate a simple analytical procedure applicable to bovine and swine urine samples for the determination of all six resorcylic acid lactones. After an enzymatic deconjugation, the urine was subjected to a one-step cleanup on a commercially available immunoaffinity chromatography cartridge. The analytes were detected by liquid chromatography-negative-ion electrospray tandem mass spectrometry using deuterium-labelled internal standards. The method was validated as a quantitative confirmatory method according to European Commission Decision 2002/657/EC. The evaluated parameters were: linearity, specificity, precision (repeatability and intra-laboratory reproducibility), recovery, decision limit, detection capability and ruggedness. The decision limits (CCalpha) obtained, were between 0.56 and 0.68 microgL(-1); recovery above 66% for all the analytes. Repeatability was between 1.4 and 5.3% and within laboratory reproducibility between 1.9 and 16.1% for the six resorcylic acid lactones. PMID- 19286012 TI - Multi-functional sample preparation procedure for measuring phytoestrogens in milk, cereals, and baby-food by liquid-chromatography tandem mass spectrometry with subsequent determination of their estrogenic activity using transcriptomic assay. AB - A method dedicated to the determination of a multiple range of phytoestrogens as endocrine disruptor compounds in infant food products was developed, with as double objective the specific measurement of 13 parameters and the evaluation of the estrogenic potency associated to this quantitative profile. A combined enzymatic and acidic chemical hydrolysis followed by a double purification on two successive C(18) and SiOH Solid Phase Extraction cartridges permitted to efficiently purify milk, cereals and baby-food samples while eliminating naturally occurring estrogen hormones. A specific liquid chromatography-tandem mass spectrometric measurement authorised unambiguous identification and quantification of the target compounds. The proposed methodology was fully validated and applied to a set of around 30 real samples, demonstrating the presence of phytoestrogens at levels globally ranging from several microgkg(-1) (ppb) to several tens mgkg(-1) (ppm). The prepared sample extracts were proven to be suitable and compatible with the evaluation of their induced biological transcriptional activity on MCF-7 cell lines. Because permitting to cope with difficult issues such as low-dose and mixture effects, this proposed methodology may appear of particular interest for further exposure assessment studies and hazard characterisation investigations related to this class of endocrine disruptor compounds. PMID- 19286013 TI - Determination of streptomycin residues in honey by liquid chromatography-tandem mass spectrometry. AB - Streptomycin is an aminoglycoside antibiotic used in apiculture to protect bees against a variety of brood diseases. Brazilian authorities have included it in the National Regulatory Monitoring Program for honey production. A simple and reliable method using liquid chromatography-tandem mass spectrometry has been developed and validated for the determination of streptomycin in honey. The chromatography separation was performed on a Gemini 5 microm C18 (50 mm x 2 mm) column using 5mM heptafluorbutiric acid/acetonitrile (85:15) as the mobile phase at a flow rate of 0.2 mLmin(-1). The detection of the analyte was achieved by positive ionization electrospray in multiple reaction-monitoring modes. Two characteristic transitions were monitored for streptomycin. Some analytical parameters were validated according to the guidelines laid down by European Commission Decision 2002/657/EC: decision limit, detection capability, recovery, precision and ruggedness. The recoveries of streptomycin from honey fortified at 2.5, 10, 15 and 20 microgkg(-1) levels are around 100%. The decision limit and detection capability of streptomycin was 3 microgkg(-1) and 4.7 microgkg(-1) respectively. PMID- 19286014 TI - The development and validation of a multiclass liquid chromatography tandem mass spectrometry (LC-MS/MS) procedure for the determination of veterinary drug residues in animal tissue using a QuEChERS (QUick, Easy, CHeap, Effective, Rugged and Safe) approach. AB - A novel rapid multiresidue/multiclass procedure with liquid chromatography tandem mass spectrometry (LC-MS/MS) detection has been developed to screen for the presence of veterinary drug residues in animal tissues. The method uses a new sample preparation procedure loosely based on QuEChERS (QUick, Easy, CHeap, Effective, Rugged and Safe) methodology. Validation to date has been restricted to chicken muscle and has been performed according to European Commission guidelines [COMMISSION DECISION of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results] for nitroimidazoles, sulphonamides, fluoroquinolones, quinolones, ionophores and dinitrocarbanilide. Recent work has shown that the method is also applicable to macrolide and lincosamide antibiotics, benzimidazoles, levamisole, avermectins and tranquillisers. PMID- 19286015 TI - Determination of 5-nitroimidazoles and corresponding hydroxy metabolites in swine kidney by ultra-performance liquid chromatography coupled to electrospray tandem mass spectrometry. AB - A rapid, sensitive and reliable multi-residue method for the simultaneous determination of four 5-nitroimidazoles (NIIMs) and their three corresponding metabolites in swine kidney was developed and validated. The compounds of interest were extracted from tissues with ethyl acetate. The crude extracts were subject to liquid-liquid partition with hexane followed by solid-phase extraction using mixed-mode strong cation-exchange column. Chromatographic separation was achieved on an AcQuity BEH C(18) column and was completed within 4 min for each injection. Data acquisition under positive electrospray tandem mass spectrometry was performed by applying multiple reaction monitoring for both identification and quantification. Mean relative recoveries from fortified samples ranged from 83% to 111%, with coefficients of variation lower than 12%. The limits of detection and quantification for the NIIMs were in the range of 0.05-0.5 and 0.1 0.5 microgkg(-1), respectively. PMID- 19286016 TI - Rapid multi-residue analysis of antibiotics in muscle by liquid chromatography tandem mass spectrometry. AB - A way of carrying out simple and rapid multi-residue analysis of antibiotics in porcine and bovine muscle by liquid chromatography-tandem mass spectrometry (LC MS/MS) is described. The method has previously been published as a screening method, but the scope has now been extended to quantification and confirmation. Nineteen compounds from five different classes of antibiotics, i.e. tetracyclines, sulfonamides, quinolones, (-lactams and macrolides, are included in the method. The samples are extracted by a single extraction using 70% methanol, diluted with water and injected in the LC-MS/MS. The run time is 7min per injection. About 60 samples can be analysed in 24h. By using this method the need for a separate screening step prior to confirmation is eliminated and consequently the total time from sampling to a confirmed result will be considerably reduced. Validation was performed according to Commission Decision 2002/657/EC. PMID- 19286017 TI - Desorption electrospray ionisation mass spectrometry: A rapid screening tool for veterinary drug preparations and forensic samples from hormone crime investigations. AB - Hormone and veterinary drug screening and forensics can benefit from the recent developments in desorption electrospray ionisation (DESI) mass spectrometry (MS). In this work the feasibility of DESI application has been studied. Using a linear ion trap or quadrupole time-of-flight (TOF) MS instrument both full-scan and data dependent collision-induced dissociation MS(n) spectra were acquired in seconds without sample preparation. Preliminary data are presented for the rapid screening of (pro)hormone supplement samples, an illegal steroid cocktail and forensic samples from veterinary drug investigations. The potential of this DESI approach is clearly demonstrated since compounds observed could be independently confirmed by liquid chromatography/TOFMS with accurate mass measurement, and/or proton nuclear magnetic resonance spectroscopy. Specific concerns related to false-positive and false-negative findings due to limitations in quantification and memory-effects are briefly discussed. It is envisaged that DESI will achieve a prominent role in hormone and veterinary drug analysis in the near future. PMID- 19286018 TI - Determination of cyanuric acid residues in catfish, trout, tilapia, salmon and shrimp by liquid chromatography-tandem mass spectrometry. AB - In May 2007, investigators discovered that waste material from the pet food manufacturing process contaminated with melamine (MEL) and/or cyanuric acid (CYA) had been added to hog and chicken feeds. At this time, investigators also learned that adulterated wheat gluten had been used in the manufacture of aquaculture feeds. Concern that the contaminated feed had been used in aquaculture and could enter the human food supply prompted the development of a method for the determination of CYA residues in the edible tissues of fish and shrimp. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed as a sensitive technique for the analysis of CYA in catfish, tilapia, salmon, trout and shrimp tissue. CYA was extracted from ground fish or shrimp with an acetic acid solution, defatted with hexane, and isolated with a graphitic carbon black solid phase extraction column. Residues were separated from matrix components using a porous graphitic carbon LC column, and then analyzed with electrospray ionization in negative ion mode on a triple quadrupole mass spectrometer. Selective reaction monitoring was performed on the [M-H](-)m/z 128 ion resulting in the product ions m/z 85 and 42. Recoveries from catfish, tilapia and trout fortified with 10-100 microgkg(-1) of CYA averaged 67% with a relative standard deviation (R.S.D.) of 18% (n=107). The average method detection limit (MDL) for catfish, tilapia and trout is 3.5 microgkg(-1). An internal standard, (13)C(3)-labeled CYA, was used in the salmon and shrimp extractions. Average recovery of CYA from salmon was 91% (R.S.D.=15%, n=18) with an MDL of 7.4 microgkg(-1). Average recovery of CYA from shrimp was 85% (R.S.D.=10%, n=13) with an MDL of 3.5 microgkg(-1). PMID- 19286019 TI - Development of a rapid method for the analysis of synthetic growth promoters in bovine muscle using liquid chromatography tandem mass spectrometry. AB - A rapid liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous identification, confirmation and quantitation of thirteen synthetic growth promoters in bovine muscle. The method was validated in accordance with the criteria defined in Commission Decision 2002/657/EC. A value of 1mugkg(-1) was chosen as the required performance level (RPL) for all analytes. The growth promoters investigated were alpha and beta trenbolone, 16-beta-OH stanozolol, methylboldenone, fluoxymesterone, methyltestosterone, medroxyprogesterone acetate, megestrol acetate, melengestrol acetate, dexamethasone, flumethasone, dienestrol and hexestrol. The method involved enzymatic hydrolysis, purification by solid phase extraction followed by analysis by UPLC-MS/MS using electrospray ionization operated in both positive and negative polarities with a total run time of 14 min. The decision limit (CCalpha) values obtained, ranged from 0.09 to 0.19 microgkg(-1) and the detection capability (CCbeta) values obtained, ranged from 0.15 to 0.32 microgkg( 1). The results of the inter-assay study, which was performed by fortifying bovine muscle samples (n=18) on three separate days, show the accuracy calculated for the various analytes to range between 98% and 102%. The precision of the method, expressed as R.S.D. values for the inter-assay variation of each analyte at the three levels of fortification (1, 1.5 and 2.0 microgkg(-1)), ranged between 3.1% and 5.8%. A Day 4 assay was carried out to examine variations due to different animals and different muscle types. PMID- 19286020 TI - Elimination kinetic of recombinant somatotropin in bovine. AB - Bovine somatotropin (bST), also called growth hormone is a protein hormone produced by the pituitary gland and responsible directly or indirectly for various effects on growth, development and reproductive functions. Its recombinant bovine somatotropin form (rbST) is used in dairy cattle to enhance milk production. Even if the effects of treatment with rbST have been largely studied, until now analytical methods able to detect rbST were limited to immunoassays, which suffer from the impossibility to distinguish between the endogenous and the recombinant form. In this study, a sample preparation procedure based on different precipitation steps, extraction on solid phase and enzymatic digestion was used to purify rbST from serum. The detection was performed by liquid chromatography coupled to tandem mass spectrometry in positive electrospray ionization mode (LC-ESI(+)-MS/MS) allowing the unambiguous identification and quantification of rbST in serum. Samples collected from a cow treated with recombinant bovine somatotropin were analysed and for the first time, the elimination kinetic specific to recombinant somatotropin has been characterized in serum. Detection of rbST was possible from 4h 30min to 4 days after administration and concentration was found up to 10ngmL(-1) during the kinetic. PMID- 19286021 TI - A determinative and confirmatory method for residues of the metabolites of carbadox and olaquindox in porcine tissues. AB - Carbadox (CBX) and olaquindox (OLQ) are used in swine feed for growth promotion, to improve feed efficiency, increase the rate of weight gain, control swine dysentery and bacterial enteritis in young swine. In 1991, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) recommended maximum residue limits (MRLs) of 30 and 5mugkg(-1) in liver and muscle tissues of pigs, respectively, based on the concentration of, and expressed as, quinoxaline-2-carboxylic acid (QCA) as marker residue. In 1998, the European Commission (EC) banned the use of CBX and OLQ in food animal production together with four other feed additives, following reports that CBX and desoxycarbadox (DCBX) are suspect carcinogens and mutagens. In 2001, the sale of CBX was halted in Canada. In 2003, JECFA recommended the withdrawal of the previously recommended acceptable daily intake (ADI) and MRLs and concluded that QCA was not a suitable marker residue for CBX, based on new sponsor studies reporting that DCBX, the suspect carcinogen, persisted in animal tissues much longer than had previously been thought. This paper presents a very sensitive LC-MS/MS method that was developed by CFIA scientists for the simultaneous determination and confirmation of DCBX residues at concentrations >/=0.050 ngkg(-1) and QCA and mQCA residues at concentrations >/=0.50 ngkg(-1)in bovine muscle, pork liver and muscle tissues. PMID- 19286022 TI - A generic method for the quantitative analysis of aminoglycosides (and spectinomycin) in animal tissue using methylated internal standards and liquid chromatography tandem mass spectrometry. AB - Aminoglycosides (AGs) are a large and diverse group of antibiotics. Although AGs may cause side effects of nephrotoxicity and ototoxicity, they are still occasionally being used for the treatment of serious infections. In this study the development of a method is described for the quantitative determination and confirmation of seven aminoglycosides (and relevant isomers) and spectinomycin in animal tissues. The extraction was based on an extraction followed by a concentration and clean-up step using weak cation exchange solid phase extraction. The separation was performed by ion-pair liquid chromatography on a C(18) column followed by mass spectrometric detection. The method was validated according to the EU requirements for a quantitative confirmatory method. Permethylated aminoglycosides (in-house synthesised internal standards) were used for accurate quantification. The accuracy of the analyses of AGs in kidney ranged from 94 to 111%, intra-day precision ranged between 2.5 and 7.4% (R.S.D.(r)) and inter-day precision ranged between 2.2 and 17.3% (R.S.D.(RL), n=21, MRL level). Accuracy (muscle tissue) varied from 83 to 128% with an intra-day precision between 2.2 and 17.3% (R.S.D.(r), n=7, MRL level). From the results it was concluded that the method was able to monitor MRL levels which ranged from 750 to 20,000 microgkg(-1) for kidney and from 50 to 10,000 microgkg(-1) for muscle tissue. PMID- 19286023 TI - Validation of multiresidue methods for veterinary drug residues; related problems and possible solutions. AB - This paper describes problems and possibly solutions encountered when validating a multiresidue method used for analyzing some 100 different veterinary drugs at trace levels in a number of matrices. The validation concept as proposed by the commission decision 2002/657/EC (CD) has been utilized. Slightly deviating approaches had to be developed in order to produce and access the data generated by a multiresidue method. This included concepts, which permit the common validation of banned and regulated substance within the same analytical method. In addition, an alternative calculation of performance parameters (e.g., CCalpha) was developed. This was done in order to ensure correct results even in the absence of measurable bank noise, as typically observed when using high resolution mass spectrometry techniques. The economical aspect of validation (number of required assays) as well as the relative importance of some performance criteria (e.g., CCbeta) have been critically discussed. PMID- 19286024 TI - In-house validation and factorial effect analysis of a liquid chromatography tandem mass spectrometry method for the determination of steroids in bovine muscle. AB - Anabolic steroids are banned from use in food-producing animals in the EU (Council Directive 96/22/EC). To control the zero-tolerance concept a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the screening and confirmation of most of the relevant natural and synthetic estrogenic and androgenic steroids in bovine muscle was developed and validated. The method permits to confirm and quantify almost all steroids below 1 microgkg(-1). The validation was carried out according to Commission Decision 2002/657/EC, chapter 3.1.3 "alternative validation", by applying a matrix-comprehensive in-house validation concept. Decision limit CCalpha, detection capability CCbeta, recovery, repeatability, within-laboratory reproducibility and the uncertainty of measurement were calculated. Furthermore, a factorial effect analysis was carried out to identify factors that have a significant influence on the method. Factors considered to be relevant for the method in routine analysis (e.g., operator, storage duration of the extracts before measurement, different cartridge lots) were systematically varied on two levels. The factorial analysis showed that different cartridge lots and different storage durations of the extracts before measurement can exert a relevant influence on the method. PMID- 19286025 TI - Determination of hormonal growth promoters in bovine hair: comparison of liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry methods for estradiol benzoate and nortestosterone decanoate. AB - The detection of steroid residues in hair is a powerful strategy to demonstrate long-term administration of these growth promoters in meat production animals. Analysis of the ester form of administered steroids is an unambiguous approach to prove the illegal use of natural hormones. For detection, gas chromatography-mass spectrometry (GC-MS/MS) was generally used. However, recent advances in liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology have improved the robustness and potential sensitivity of this method. This paper describes development and validation, according to Commission Decision 2002/657/EC, of LC MS/MS and GC-MS/MS methods, in two separate laboratories, for determination of steroid esters in bovine hair. Bovine hair samples taken from animals treated with estradiol-3-benzoate and nortestosterone decanoate, as well as from an untreated animal, were used to evaluate the comparability of the results of the two validated methods. The results of the inter-comparison demonstrate that both the LC-MS/MS and the GC-MS/MS methods are fit for purpose and capable of determining steroid esters in hair samples from treated bovine animals. PMID- 19286026 TI - Depletion of chloramphenicol in trout after a hypothetic therapeutic treatment. AB - The study was intended to evaluate the depletion of chloramphenicol (CAP) in rainbow trout (about 300-550 g body weight), after 10 days treatment with fish feedstuff containing chloramphenicol. A total of 60 animals were separated in two groups: one was fed with CAP containing feedstuff in order to have a dosage of about 80 mgkg(-1)day(-1), while a second group of fishes was fed with feedstuff not containing any CAP formulation (negative controls). The treatment was maintained for 10 days. After this period, groups of 2-5 animals were sacrificed at different withdrawal times up to a maximum of 31 days. Muscle tissues of each group of animals were then analysed for quantitative residual CAP determination both by enzyme linked immunoassay (ELISA) and liquid chromatography coupled to mass spectrometry (HPLC/MSMS). The methods applied were in house validated according to the guidelines laid down by the European Decision 657/2002/EC. Results and considerations are presented. PMID- 19286027 TI - Detection and identification of 20-hydroxyecdysone metabolites in calf urine by liquid chromatography-high resolution or tandem mass spectrometry measurements and establishment of their kinetics of elimination after 20-hydroxyecdysone administration. AB - Ecdysteroids, which are steroid hormones in invertebrates, but are also present in plants, could be potentially used as anabolic agents in food-producing animals because of their growth-promoting properties. In this context, the metabolism of 20-hydroxyecdysone (20E) has been investigated in cattle in order to efficiently control its potential misuse. The analytical procedure involves purification on two solid-phase extraction cartridges (SPE octadecylsilyl and SPE silica) prior to detection based on liquid chromatography coupled to high resolution mass spectrometry in negative electrospray ionization mode (LC-(ESI-)-HRMS). Each new signal appearing on full-scan HRMS (30,000) during the analysis was investigated by tandem mass spectrometry (MS/MS). Comparison of the mass spectra pattern between 20E and potential metabolites has given informations on the chemical structures of the metabolites. This targeted approach, combining HRMS and MS(n) measurements on a linear trap in tandem with an orbital trap, allowed us to elucidate the structure of several 20E metabolites in calf urine: 14-deoxy-20 hydroxyecdysone, 20,26-dihydroxyecdysone and 14-deoxy-20,26-dihydroxyecdysone, the last of which had never previously been reported in bovine. The kinetics of elimination of these metabolites were investigated, and two of them were monitored by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) measurements over 6 days after 20E administration to calves, thus increasing the time-window for detection after 20E administration and thereby allowing for more efficient control of its misuse. PMID- 19286028 TI - Rapid method for the determination of tranquilizers and a beta-blocker in porcine and bovine kidney by liquid chromatography with tandem mass spectrometry. AB - A fast and simple liquid chromatography with tandem mass spectrometry method for detection and confirmation of tranquilizers (chlorpromazine, propionylpromazine, acepromazine, triflupromazine, promazine, azaperone and its metabolite, azaperol) and beta-blocker (carazolol) in porcine and bovine kidney has been presented. The method relies on the extraction with acetonitrile followed by centrifugation. After evaporation of acetonitrile, the residue was reconstituted in a mobile phase and filtrated. The separation of analytes was performed on a C18 column using a mobile phase of acetonitrile and ammonium formate buffer (0.05 M, pH 4.5) with gradient elution. The electrospray ionization was used to obtain the protonated molecules [M+H](+) and two product ions were monitored for each compound. For quantification deutered internal standards were used. The whole method has been validated according to the European Union requirements. Specificity, decision limit (CCalpha), detection capability (CCbeta), trueness and precision were determined. The results showed good trueness ranged from 73.2% to 110.6% with a good R.S.D., less than 13.0% under within-laboratory reproducibility conditions. The calculated critical concentrations of CCalpha for phenothiazines were between 5.8 and 6.6 microgkg(-1) while for azaperone CCalpha was 105.5 microgkg(-1) and for azaperol was 121.4 microgkg(-1). CCalpha for carazolol was 16.7 microgkg(-1) in bovine and 21.9 microgkg(-1) in porcine kidney. CCbeta for phenothiazines were between 6.3 and 7.6 microgkg(-1), for azaperone was 119.0 microgkg(-1) and for azaperol was 140.0 microgkg(-1). For carazolol in bovine kidney CCbeta was 18.6 microgkg(-1) whereas in porcine kidney was 24.4 microgkg(-1). PMID- 19286029 TI - How to decrease ion suppression in a multiresidue determination of beta-agonists in animal liver and urine by liquid chromatography-mass spectrometry with ion trap detector. AB - A new liquid chromatography-mass spectrometry with ion-trap detection method for the determination of clenbuterol, ractopamine, clenpenterol, brombuterol, hydroxyclenbuterol, mapenterol and mabuterol in animal liver and urine is described to decrease ion suppression in both matrices. The developed method consists of an aqueous extraction and a two-step clean-up. Final extract was injected in a liquid chromatograph with ion trap mass spectrometer optimized to obtain MS(3) ions which provided more specificity and better signal to noise ratio in the investigated analytes. Those steps have been essential to reach decision limits (CCalpha) values below the proposed MRPL values for each substance. The CCalpha were determined and their values were 0.05 microgkg(-1) for clenbuterol, 0.2 microgkg(-1) for ractopamine and 0.1 microgkg(-1) for the other analytes in liver and the same values in microgL(-1) for urine samples. PMID- 19286030 TI - New method for the analysis of flukicide and other anthelmintic residues in bovine milk and liver using liquid chromatography-tandem mass spectrometry. AB - A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) multi-residue method for the simultaneous quantification and identification of 38 residues of the most widely used anthelmintic veterinary drugs (including benzimidazoles, macrocyclic lactones, and flukicides) in milk and liver has been developed and validated. For sample preparation, we used a simple modification of the QuEChERS method, which was initially developed for pesticide residue analysis. The method involved extracting sample (10 g) with acetonitrile (10 mL), followed by phase separation from water (salting out) with MgSO(4):NaCl (4:1, w/w). After centrifugation, an aliquot of the extract (1 mL) was purified by dispersive solid phase extraction with MgSO(4) (150 mg) and C(18) (50mg), prior to LC-MS/MS analysis. Two injections of the same extract were required with the LC-MS/MS instrument to cover the 30 electrospray positive and 8 electrospray negative analytes. The limit of quantitation of the method was 5 microgkg(-1) for 37 analytes (and 10 microgkg(-1) for dichlorvos). The method was successfully validated according to the 2002/657/EC guidelines. Recovery of analytes was typically in the 70-120% range, with repeatabilities and reproducibilities typically <15% in milk and <20% in liver. PMID- 19286031 TI - Simple and rapid determination of enrofloxacin and ciprofloxacin in edible tissues by turbulent flow chromatography/tandem mass spectrometry (TFC-MS/MS). AB - A simple and rapid method for the determination of residues of enrofloxacin and its metabolite ciprofloxacin in edible tissues of farm animals using turbulent flow chromatography/tandem mass spectrometry (TFC-MS/MS) is described. The tissue samples were extracted with a mixture of acetonitrile, water and formic acid. After addition of internal standard solution, an aliquot of the extract was injected into the turbulent flow chromatography system. Matrix components contained in the injected sample were separated from the retained analytes on a polymer-based extraction column suited for pretreatment of samples at high flow rates. The analytes were eluted to an analytical column and the quantitative determination was performed using a tandem mass spectrometric detector. The run time for the analysis was 4 min. The limit of quantitation (LOQ) was 25 microgkg( 1) for each analyte and tissue material. Validation was performed in edible tissues of cattle, pig, turkey and rabbit in the range from 25 microgkg(-1) to at least twice the MRL. Mean recovery rates for the tissues of the different species were in the range from 72 to 105% with a coefficient of variation (CV) between 4.3 and 18%. PMID- 19286032 TI - Determination of ten sulphonamides in egg by liquid chromatography-tandem mass spectrometry. AB - A precise and reliable method for the determination of 10 sulphonamide antibiotics (sulfadiazine, sulfathiazole, sulfamerazine, sulfamethazine, sulfamethoxypyridazine, sulfachloropyridazine, sulfamethoxazole, sulfamonomethoxine, sulfadimethoxine and sulfaquinoxaline) in egg by liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed. Drugs were extracted using a mixture of dichloromethane/acetone (50:50, v/v), acidified with acetic acid and then cleaned-up on a cation-exchange solid-phase extraction (SPE) cartridge. The chromatographic separation was performed by gradient on a C(18) column with a mobile phase of methanol-water containing 0.1% formic acid and 5mM ammonium acetate, then sulphonamides were detected in a triple-quadrupole mass spectrometer operated in positive electrospray ionization mode (ESI(+)). The method was validated at 15, 30 and 45 microgkg(-1). These levels were much lower than the corresponding maximum residue limit of 100 microgkg(-1) set for sulphonamides in several matrices but not in eggs, where the presence of such residues is not permitted. Results were quantitated against the selected internal standard (13)C(6)-sulphamethazine and also according to the matrix-matched approach. The within-laboratory reproducibility, expressed as a relative standard deviation, never exceeded 21%. All decision limit (CCalpha) values lied in the range between 16.1 and 20.5 microgkg(-1) and the corresponding results for detection capability (CCbeta) were 16.9 and 25.7 microgkg(-1). Ruggedness was estimated according to the Youden robustness test. PMID- 19286033 TI - Determination of dapsone in meat and milk by liquid chromatography tandem mass spectrometry. AB - Within the EU the use of dapsone (4,4-diaminodiphenylsulfone) is prohibited in food-producing animals and consequently it's included in the Annex IV of the Directive 90/2377/EC. A quantitative confirmatory method has been developed and validated according to the criteria defined in the Commission Decision 2002/657/EC, for the determination of dapsone in meat and milk. Samples, after homogenization in alkaline conditions and organic solvent extraction, were purified on silica gel solid phase extraction cartridges. The eluate was evaporated and redissolved in mobile phase and was analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS) in positive electrospray ionisation (ESI) using deuterium labelled Sulphadimidine-d7 as internal standard. The calculated value for, decision limit, CCalpha is 0.12 microgkg(-1), and the detection capability; CCbeta value is 0.16 microgkg(-1). PMID- 19286034 TI - Validation and application of a yeast bioassay for screening androgenic activity in calf urine and feed. AB - Bioassays are valuable tools for combating the illegal use of steroids in cattle fattening. Previously we described the construction and properties of a rapid and robust yeast androgen bioassay stably expressing the human androgen receptor (hAR) and yeast enhanced green fluorescent protein (yEGFP), the latter in response to androgens. In the present study this yeast androgen bioassay was validated as a qualitative screening method for the determination of androgenic activity in calf urine and animal feed. This validation was performed according to EC Decision 2002/657. 20 blank samples were spiked with testosterone, 17alpha methyltestosterone, 19-nortestosterone, 17beta-trenbolone, 17beta-boldenone or 17alpha-methylboldenone at 2 or 15 ngmL(-1) in urine and 50 or 100 ngg(-1) in feed. All blank and spiked samples fulfilled the CCalpha and CCbeta criterions, meaning that all 20 blank samples gave signals below the determined decision limits CCalpha and were thus classified as compliant (alpha=1%). For each component, at least 19 out of the 20 spiked samples gave a signal above the CCalpha and were thus classified as suspect (beta=5%). The method was specific, and high amounts of dexamethasone did not interfere with the outcome of the test. Although high levels of 17alpha-ethynylestradiol can significantly inhibit the response obtained with low amounts of androgens, that situation is not relevant in veterinary practice. When stored at their specific conditions, the androgens in feed were stable for at least 91 days. Real urine samples from a national control program were screened and a representative part of the compliant and suspect samples were confirmed by gas chromatography-tandem mass spectrometry. PMID- 19286035 TI - Biosensor-based detection of reduced sex hormone-binding globulin binding capacities in response to growth-promoter administrations. AB - Growth-promoting agents are illicitly used during animal rearing processes and the detection of their use is limited by new compounds and dosing practices that limit the efficiency of current testing which is based on residue analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography mass spectrometry (GC-MS) methodology. An alternative approach is to use indirect biological evidence as a screening tool to identify growth-promoter treated animals thus improving the effectiveness of residue testing through the targeted sampling of these animals. Sex hormone-binding globulin (SHBG) is a glycoprotein which binds and controls the levels of sex-hormones within the circulation. Using a biosensor assay based on measurement of binding to an immobilised 1alpha dihydrotestosterone (1alpha-DHT) derivative, reduced SHBG binding capacities were detected in growth-promoter treated animals. During the course of a veal treatment regime based on repeated oestradiol benzoate, nortestosterone decanoate and dexamethasone administrations, treated male and female calves were shown to have significantly lower SHBG capacities. To assess the effectiveness of using SHBG binding capacities as a biomarker of treatment and to investigate the role of individual growth-promoter components to the SHBG capacity lowering effects, adult heifer animals were subjected to repeated doses of nortestosterone decanoate. These animals also demonstrated a reduction in SHBG capacity levels at Day 39 of the study, in contrast to oestradiol benzoate treated adult steers who were found to have unaltered levels. These findings suggest that the measurement of SHBG binding capacities using a biosensor assay has potential in the identification of illegally treated animals, particularly those exposed to androgens. PMID- 19286036 TI - A rapid microbial inhibition-based screening strategy for fluoroquinolone and quinolone residues in foods of animal origin. AB - A rapid, high-throughput antimicrobial screening assay for the detection of fluoroquinolone and 4-quinolone residues in foods of animal origin has been developed in ampoule format. The assay employs a single Escherichia coli species sensitive to those Gram-negative inhibitiory antimicrobial compounds and is presented in a comparable format to the existing commercially available Premi Test and Delvotest ampoule-based microbial inhibition tests (DSM, Delft, The Netherlands). In the novel E. coli assay the microorganism, in vegetative state, is inoculated into a nutrient agar pellet containing a pH sensitive acid-base indicator dye. A simple extraction protocol that is selective for fluoroquinolone and quinolone compounds was developed to recover, cleanup and concentrate the target analyte(s) from a variety of tissue types and matrices prior to screening analysis. The method detected 16 target compounds at concentrations equal to or below the maximum residue limits (where applicable). The method has been validated using the prototype assay in accordance with the 2002/657/EC guidelines for the validation of qualitative screening assays. False positive and false negative responses rates for the procedure have been determined as less than 5%. The stability of a selection of representative target analytes has been demonstrated for a 20-week period under a variety of storage conditions both in tissue and in extract. PMID- 19286037 TI - Detection of anabolic androgenic steroid abuse in doping control using mammalian reporter gene bioassays. AB - Anabolic androgenic steroids (AAS) are a class of steroid hormones related to the male hormone testosterone. They are frequently detected as drugs in sport doping control. Being similar to or derived from natural male hormones, AAS share the activation of the androgen receptor (AR) as common mechanism of action. The mammalian androgen responsive reporter gene assay (AR CALUX bioassay), measuring compounds interacting with the AR can be used for the analysis of AAS without the necessity of knowing their chemical structure beforehand, whereas current chemical-analytical approaches may have difficulty in detecting compounds with unknown structures, such as designer steroids. This study demonstrated that AAS prohibited in sports and potential designer AAS can be detected with this AR reporter gene assay, but that also additional steroid activities of AAS could be found using additional mammalian bioassays for other types of steroid hormones. Mixtures of AAS were found to behave additively in the AR reporter gene assay showing that it is possible to use this method for complex mixtures as are found in doping control samples, including mixtures that are a result of multi drug use. To test if mammalian reporter gene assays could be used for the detection of AAS in urine samples, background steroidal activities were measured. AAS-spiked urine samples, mimicking doping positive samples, showed significantly higher androgenic activities than unspiked samples. GC-MS analysis of endogenous androgens and AR reporter gene assay analysis of urine samples showed how a combined chemical-analytical and bioassay approach can be used to identify samples containing AAS. The results indicate that the AR reporter gene assay, in addition to chemical-analytical methods, can be a valuable tool for the analysis of AAS for doping control purposes. PMID- 19286038 TI - Improved screening method for the detection of a range of nitroimidazoles in various matrices by optical biosensor. AB - An immunobiosensor assay was developed for the multi-residue screening of a range of nitroimidazole compounds in various species and sample types including porcine, bovine and ovine kidney, avian liver, serum and eggs and bovine milk. A polyclonal antibody which binds at least seven of the major nitroimidazoles and their metabolites was raised in a sheep after inoculation with a metronidazole protein conjugate. Sample homogenates were extracted into acetonitrile and subjected to micro-centrifugation prior to biosensor analysis. Validation data obtained from the analysis of 20 fortified samples has shown that the method has a detection capability (CCbeta) of less than 1 microgkg(-1) (or microgL(-1)) for dimetridazole (DMZ) in all species and matrices investigated. In addition, cross reactivity data and the analysis of a small number of fortified samples have shown that the method will also detect a range of other major parent nitroimidazoles and their metabolites including ronidazole (RNZ), ipronidazole (IPZ), metronidazole (MNZ), hydroxymetronidazole (MNZOH), hydroxydimetridazole (DMZOH) and hydroxyipronidazole (IPZOH). The cross-reactivity profile and validation data for the detection of these nitroimidazoles are presented together with the results obtained following the analysis of a small number of incurred samples using the developed method. PMID- 19286039 TI - Inter-laboratory comparison of a yeast bioassay for the determination of estrogenic activity in biological samples. AB - An inter-laboratory exercise was performed with a yeast estrogen bioassay, based on the expression of yeast enhanced green fluorescent protein (yEGFP), for the determination of estrogenic activity in extracts of calf urine samples. Urine samples were spiked with 1 and 5 ngmL(-1) 17beta-estradiol and 17alpha ethynylestradiol, 10 and 50 ngmL(-1) mestranol, and 100 ngmL(-1) testosterone and progesterone. Sample extracts of blank and spiked urine samples were prepared at our laboratory and sent to seven laboratories together with a reagent blank, a DMSO blank, and eight 17beta-estradiol stock solutions in DMSO ranging in concentration from 0 to 545 ngmL(-1). Sample extracts and standards were coded and tested blindly. A decision limit (CCalpha) was determined based on the response of seven blank urine samples. Signals of the negative controls, e.g. urine samples spiked with 100 ngmL(-1) testosterone or progesterone, were all below the determined CCalpha and were thus screened as compliant. Positive controls, i.e. the urine samples spiked at two levels with 17beta-estradiol, 17alpha-ethynylestradiol and mestranol, were almost all screened as suspect, i.e. gave signals above the determined CCalpha. Determined EC(50) values calculated from the 17beta-estradiol dose-response curves obtained by the seven laboratories ranged from 0.59 to 0.95 nM. PMID- 19286040 TI - Validation of an enzyme-linked immunosorbent assay screening for quinolones in egg, poultry muscle and feed samples. AB - Quinolones are a group of chemotherapeutic agents with an excellent efficiency against poultry pathogens. Two commercial enzyme-linked immunosorbent assay (ELISA) tests have been applied in parallel for the qualitative screening analysis of several quinolones in eggs, poultry muscle and feeds at the required levels. During the validation study, carried out according to the Commission Decision 2002/657/EC criteria, two different sample treatments were compared in foods: the simple and fast procedure suggested by the kit producer (Euro Diagnostica) and a more complex solid-phase extraction (SPE) sample preparation. The results demonstrated that the method based on SPE clean up exhibited better characteristic performances, particularly in eggs for which lower detection levels are required. Despite the fact that screening methods should be rapid and cheap, the use of non-chromatographic techniques such as ELISA for multiresidual detection of a class of substances involves some additional attention to sample preparation. PMID- 19286041 TI - Multiresidue method for the triphenylmethane dyes in fish: Malachite green, crystal (gentian) violet, and brilliant green. AB - Liquid chromatographic methods are presented for the quantitative and confirmatory determination of crystal violet (CV; also known as gentian violet), leucocrystal violet (LCV), brilliant green (BG), and leucobrilliant green (LBG) in catfish. LCV and LBG were oxidized to the chromic CV and BG by reaction with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, and residues were measured as the combined CV+/-LCV and BG+/-LBG. These methods are extensions of published methods for malachite green (MG) analysis to allow simultaneous determination of MG, CV, and BG. Residues were extracted from muscle with ammonium acetate buffer and acetonitrile, and extracts cleaned up using dichloromethane partitioning and solid-phase extraction. Extracts were analyzed by liquid chromatography with visible detection (LC-VIS). The method was validated for catfish fortified with LCV over the range 0.25-10 ngg(-1) and CV at 2 ngg(-1). Average recoveries were 90.6% (+/-8.1% R.S.D., n=45) for LCV and 84.4% (+/-4.2% R.S.D., n=6) for CV. The average recovery for samples fortified with BG or LBG over the range 0.5-10 ngg( 1) was 67.2% (+/-14.8% R.S.D., n=31). CV and BG were confirmed in fish extracts by ion trap LC-mass spectrometry (LC-MS(n)) with no discharge-atmospheric pressure chemical ionization. Average LC-MS(n) recoveries were 96.5, 96.6, and 70.2% for samples fortified with CV, LCV, and BG or LBG. The limits of detection for CV, BG, and MG were in the range of 0.07-0.24 ngg(-1) (ppb) for the two different instrumental methods. This methodology was applied to the analysis of catfish treated with CV and BG. PMID- 19286042 TI - Comparison of screening methods for antibiotics in beef kidney juice and serum. AB - Rapid screening tests can be used as part of an efficient program designed to monitor veterinary drug residues in cattle. In this work, three rapid tests designed to screen samples for the presence of antibiotic residues, the Fast Antimicrobial Screen Test (FAST), Premi and Kidney Inhibition Swab (KIS) tests, were compared using beef kidney juice and serum samples. In order to provide a realistic assessment, potentially incurred samples of beef kidney juice and serum were obtained from 235 carcasses which had been retained by inspectors in a processing plant for further testing. In addition, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was conducted on these samples to identify what antibiotics were present, if any, and their levels. The comparison of the three rapid screening test results with those from LC-MS/MS analysis allowed for a more complete comparison of the relative sensitivity of these analytical methods, as well as valuable information on false positive and negative response rates. PMID- 19286043 TI - Comparison of three microbial screening methods for antibiotics using routine monitoring samples. AB - Monitoring large numbers of slaughter animals for the presence of antimicrobial residues is preferably carried out using microbiological screening methods, because of their high cost-effectiveness. An evaluation of the Nouws antibiotic test (NAT) was performed on routine monitoring samples and the performance of the method was compared with two other microbial screening methods: Screening test for antibiotic residues (STAR) and Premi Test. Analysis of 591 samples yielded four MRL violations. Three of them concerned tetracyclines that were only detected with the NAT and the STAR method. The fourth, 172 microgkg(-1) Sulfadiazine, was detected by all three methods. Additionally, 156 microgkg(-1) Tulathromycin was found in porcine meat, while for this residue no MRL in muscle has been established. PMID- 19286044 TI - Detection of anabolic steroids in dietary supplements: the added value of an androgen yeast bioassay in parallel with a liquid chromatography-tandem mass spectrometry screening method. AB - Recently we constructed a recombinant yeast cell that expresses the human androgen receptor (hAR) and yeast enhanced green fluorescent protein (yEGFP), the latter in response to androgens. When exposed to testosterone, the concentration where half-maximal activation is reached (EC(50)) was 50 nM. Eighteen different dietary supplements, already analysed by a liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for the presence of anabolic steroids, were screened for androgenic activity. Eleven samples containing at least one anabolic steroid, with a concentration that was around or above 0.01 mgunit(-1) according to LC-MS/MS, were also positive in the bioassay. Seven samples did not contain any of the 49 compounds screened for in LC-MS/MS. In contrast two of them were positive in the bioassay. Bioassay-directed identification, using the bioassay as an off-line LC-detector and LC-time of flight-MS with accurate mass measurement was carried out in these two samples and revealed the presence of 4-androstene 3beta,17beta-diol and 5alpha-androstane-3beta,17beta-diol in the first and 1 testosterone in the second supplement, showing the added value of the bioassay in comparison with a LC-MS/MS screening method alone. PMID- 19286045 TI - Study of the depletion of lincomycin residues in honey extracted from treated honeybee (Apis mellifera L.) colonies and the effect of the shook swarm procedure. AB - Bee colonies were treated with 1.2g lincomycin hydrochloride per hive (single treatment in sucrose solution) and samples of honey were then collected at intervals over a 41-week period. The samples were analysed for lincomycin using Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS). The highest mean concentration of lincomycin (pooled analytical results for brood and super honey) was 24 microugg(-1) 3 days after treatment, a mean of 3.5 microgg(-1) after 129 days. The shook swarm procedure was investigated and resulted in a lincomycin concentration of 34 microgg(-1) in honey (pooled results for brood and super honey) 3 days after treatment, declining to 0.38 microgg(-1) 129 days after treatment. Lincomycin was persistent in the hive and detected in all over winter (290 days after dosing) samples of honey collected from both non-shook swarmed and shook swarmed colonies. The results overall indicate that lincomycin parent is a suitable marker compound to detect lincomycin misuse in apiculture. PMID- 19286046 TI - Detection of glucocorticoid bioactivity in bovine urine samples using a reporter gene assay. AB - The illegal use of anabolic substances in the meat producing industry is an ongoing problem due to the continual production of new synthetic compounds and/or the practice of low-level cocktail administration to avoid detection by the surveillance schemes of EU member states National Plan surveillance systems. We present a highly sensitive reporter gene assay and sample extraction procedure based on a two step solid phase extraction and high performance liquid chromatography, developed for the detection of glucocorticoid abuse in bovine urine. The assay is capable of detecting compounds with glucocorticoid bioactivity and is extremely sensitive with an EC(50) of 0.79 ngmL(-1) for dexamethasone. New or unknown compounds with glucocorticoid bioactivity and low level cocktail mixtures are detectable by this assay. Cross-reactivity data for a range of 11beta-hydroxyglucocorticoids has been provided. This assay shows low interference from the 11-keto prohormones and other steroidal hormones. The assay may be suitable for application in other matrices such as hair. In conclusion this screening assay offers advantages over existing analytical techniques. PMID- 19286047 TI - Antibody production: Low dose immunogen vs. low incorporation hapten using salmeterol as a model. AB - Haptens are low molecular weight compounds that are non-immunogenic and so must be conjugated to carrier molecules to elicit an immune response. Doses of 50-1000 microg protein conjugate have been suggested for immunisation of rabbits with hapten-protein immunogens. Although larger doses may give a faster response, lower doses may result in higher affinity antibodies. The amount of hapten presented to the host's immune system can be controlled by varying either the amount of immunogen administered or the quantity coupled to a fixed amount of protein. This study compares the two approaches for the production of antibodies to the beta-agonist salmeterol as a model. A range of salmeterol-HSA conjugates was prepared, with varying molar ratios of hapten:protein (80:1, 15:1, 7.5:1, 4:1), for use as immunogens. The 80:1 immunogen was administered to different animals at four concentrations (0.5, 0.2, 0.1 and 0.05 mgdose(-1)) while the remaining three immunogens were administered at 0.5 mgdose(-1). The effects of immunogen dose and hapten incorporation on the titre and affinity of the antibodies produced to salmeterol were investigated. It was found that both approaches resulted in the production of more sensitive antibodies, although reducing the degree of hapten incorporation brought about a larger reduction in antibody titre. Reducing the degree of hapten incorporation also produced a more consistent pattern of results regarding antibody sensitivity (after the sixth immunisation in this study) which may make it easier to predict the most suitable time for antibody harvesting. PMID- 19286048 TI - A National Residue Control Plan from the analytical perspective--the Brazilian case. AB - Food safety is a strategic topic entailing not only national public health aspects but also competitiveness in international trade. An important component of any food safety program is the control and monitoring of residues posed by certain substances involved in food production. In turn, a National Residue Control Plan (NRCP) relies on an appropriate laboratory network, not only to generate analytical results, but also more broadly to verify and co-validate the controls built along the food production chain. Therefore laboratories operating under a NRCP should work in close cooperation with inspection bodies, fostering the critical alignment of the whole system with the principles of risk analysis. Beyond producing technically valid results, these laboratories should arguably be able to assist in the prediction and establishment of targets for official control. In pursuit of analytical excellence, the Brazilian government has developed a strategic plan for Official Agricultural Laboratories. Inserted in a national agenda for agricultural risk analysis, the plan has succeeded in raising laboratory budget by approximately 200%, it has started a rigorous program for personnel capacity-building, it has initiated strategic cooperation with international reference centres, and finally, it has completely renewed instrumental resources and rapidly triggered a program aimed at full laboratory compliance with ISO/IEC 17025 requirements. PMID- 19286049 TI - Food flavonoid aryl hydrocarbon receptor-mediated agonistic/antagonistic/synergic activities in human and rat reporter gene assays. AB - Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor mediating the adverse effects of dioxins and polycyclic aromatic hydrocarbons (PAHs). In this study, we investigated the genetic-, time-, dose-, species- and tissue-dependent AhR-mediated agonistic/antagonistic activities of three food flavonoids: quercetin, chrysin and genistein. To that end, four stably transfected cell lines were used in cell-based luciferase reporter gene assays: three lines were transformed with the ptKLuc vector harbouring four dioxin responsive elements (DREs) upstream of the thymidine kinase promoter and the luciferase gene (HepG2-Luc, T-47D-Luc and H4IIE-ULg). The fourth is a patented cell line transformed with a different construct: H4IIE DR-CALUX((R)). Both H4IIE cells were compared for their genetic construction. Human hepatoma (HepG2-Luc) and human breast tumour (T-47D-Luc) cells were compared for tissue-dependent effects. Rat hepatoma (H4IIE-ULg) and human hepatoma (HepG2-Luc) cells were compared for species-dependent activities. We concluded that quercetin, chrysin and genistein act in a time-, dose-, species- and tissue-specific way. For example, genistein displayed agonistic activities when exposed to rat hepatoma cells during 6h but not after 24h. Flavonoids displayed agonistic/antagonistic activities in human breast tumour cells, depending on the exposure time, while in human hepatoma cells, only antagonistic activities of flavonoids were measured. In addition, we report, in all the cells, a synergy between an isoflavone and two food contaminants; the 2,3,7,8-tetrachlorodibenzo-p-dioxin and 3 methylcholanthrene, a PAH. In rat cells, this synergy occurred when cells were exposed to flavonoids and contaminant for 6h, while it was observed in human cells only after 24h. PMID- 19286050 TI - In-house reference materials: 5-hydroxyflunixin and meloxicam in cow milk preparation and evaluation. AB - Reference materials are helpful to evaluate the performance of laboratories as well as being useful for the quality control of analytical procedures. Certified reference materials and other reference materials containing non-steroidal anti inflammatory drugs in milk are however, not available. Therefore, production and evaluation of in-house reference materials with incurred residues of 5 hydroxyflunixin (5OHFLU) and meloxicam (MEL) in cow milk has been performed. The milk was collected 12h after dosing from cows which received meloxicam (0.5 mgkg( 1) b.w., i.v., single dose) or flunixin meglumine (2.2 mgkg(-1) b.w., i.v. during three days). The concentrations of analytes were checked in the milk samples. The milk was diluted with milk free from NSAIDs residues, homogenised, put into sterile 20 mL vials, frozen and lyophilised. The vials were weighed before and after lyophilisation, in order to calculate the amount of water necessary for reconstitution, and were stored at a temperature of -20+/-2 degrees C. For the homogeneity study, 10 random samples were analysed in duplicate and the results were interpreted using Cochran's test, Horwitz standard deviation and the test for a sufficient homogeneity. The assigned values, calculated from the results of the homogeneity test were 54.3 microgkg(-1) for 5OHFLU and 46.4 microgkg(-1) for MEL. The samples were tested for their stability every 14 days for 2 months and after 9 months. It has been confirmed that an appropriate homogeneity and stability of the produced in-house reference material has been obtained. PMID- 19286051 TI - Effect of growth-promoting 17beta-estradiol, 19-nortestosterone and dexamethasone on circulating levels of nine potential biomarker candidates in veal calves. AB - The use of screening methods based on the detection of biological effects of growth promoters is a promising approach to assist residue monitoring. To reveal useful effects on protein metabolism, male and female veal calves at 10 weeks of age were treated thrice with a combination of 25mg 17beta-estradiol 3-benzoate and 150 mg 19-nortestosterone decanoate with 2 weeks intervals and finally once with 4 mg dexamethasone. Hormone-treated calves showed a significant accelerated growth rate over 6 weeks. Plasma samples of treated and control calves were analysed for immunoreactive inhibin (ir-inhibin), osteocalcin, insulin-like growth factor 1 (IGF-1), insulin-like growth factor-binding protein 2 (IGFBP-2), IGFBP-3, luteinzing hormone (LH), follicle-stimulating hormone (FSH) and prolactin using immunoaffinity assays. Hormone treatment did not affect levels of IGF-1, IGFBP-2, IGFBP-3, LH, FSH and prolactin. The concentration of circulating ir-inhibin decreased, however, significantly (P<0.05) in bull calves upon administration of the sex steroids, whereas it remained unchanged in the female animals. Dexamethasone treatment decreased significantly (P<0.05) circulating levels of osteocalcin in both female and male animals. Ir-inhibin and osteocalcin were, therefore, considered as candidates for a protein biomarker-based screening assay for detection of abuse of estrogens, androgens and/or glucocorticoids in cattle fattening, which is being developed in the framework of EU research project BioCop (www.biocop.org). PMID- 19286052 TI - Adiponectin levels in patients undergoing coronary stenting. PMID- 19286053 TI - Cosmetic procedures for the primary care physician. Foreword. PMID- 19286054 TI - Complications of botulinum neurotoxin. PMID- 19286055 TI - Varicose veins. PMID- 19286056 TI - Chemical peels, dermabrasion, and laser therapy. PMID- 19286057 TI - Skin fillers. PMID- 19286058 TI - George Duncan special issue. PMID- 19286059 TI - Health governance: law, regulation and policy. PMID- 19286060 TI - The role of public health governance. Foreward. PMID- 19286061 TI - No more business as usual: enticing companies to sharply lower the public health costs of the products they sell. AB - Cigarettes, alcohol, junk food and motor vehicles cause a staggeringly high level of death, injury and disease. Business leaders from the industries that make these products currently try to frame these negative outcomes as 'collateral damage' that is someone else's problem. That framing is not only morally objectionable, but also overlooks the possibility that, with proper prodding, industry could substantially mitigate these public health disasters. A promising regulatory tool called 'performance-based regulation' is a new approach to combating the problem. Simply put, performance-based regulation would impose a legal obligation on manufacturers to reduce their negative social costs. Rather than suing the firms for damages, or telling them how they should run their businesses differently (as typical 'command and control' regimes do), performance based regulation allows the firms to determine how best to decrease today's negative public health consequences. Like other public health strategies, performance-based regulation shifts the focus away from individual consumers on to those who are far more likely to achieve real public health gains. Analogous to a tax on causing harm that exceeds a threshold level, performance-based regulation seeks to harness private initiative in pursuit of the public good. PMID- 19286062 TI - [Abstracts from the French-speaking Meeting of Neurology, 1-4 April 2009, Lille, France]. PMID- 19286063 TI - Cardiologists should be less passive about smoking cessation. PMID- 19286064 TI - Listening to patients with rare diseases. PMID- 19286065 TI - Shining a spotlight on glaucoma. PMID- 19286066 TI - The bioabsorbable stent as a virtual prosthesis. PMID- 19286067 TI - Effectiveness of PCI for non-acute coronary artery disease. PMID- 19286068 TI - The ischaemia/bleeding balance in PCI. PMID- 19286069 TI - Secondary cardiovascular risk prevention--we can do better. PMID- 19286070 TI - Threats to the future of cardiovascular research. PMID- 19286071 TI - Home blood-pressure monitoring: US and European consensus. PMID- 19286073 TI - Crisis. PMID- 19286075 TI - Israel-Gaza conflict. PMID- 19286076 TI - Israel-Gaza conflict. PMID- 19286077 TI - Israel-Gaza conflict. PMID- 19286078 TI - Israel-Gaza conflict. PMID- 19286079 TI - Israel-Gaza conflict. PMID- 19286080 TI - Israel-Gaza conflict. PMID- 19286081 TI - Antenatal corticosteroids to prevent preterm birth. PMID- 19286083 TI - Trade and health: dealing in death. PMID- 19286084 TI - Antenatal corticosteroids to prevent preterm birth. PMID- 19286085 TI - Achieving South Africa's National Strategic Plan for HIV/AIDS. PMID- 19286086 TI - Homelessness and soul murder. PMID- 19286087 TI - The British Medical Association--a toothless lion. PMID- 19286088 TI - Medical ethics and prisoners. PMID- 19286089 TI - A bioabsorbable everolimus-eluting coronary stent system (ABSORB): 2-year outcomes and results from multiple imaging methods. AB - BACKGROUND: Drug-eluting metallic coronary stents predispose to late stent thrombosis, prevent late lumen vessel enlargement, hinder surgical revascularisation, and impair imaging with multislice CT. We assessed the safety of the bioabsorbable everolimus-eluting stent (BVS). METHODS: 30 patients with a single de-novo coronary artery lesion were followed up for 2 years clinically and with multiple imaging methods: multislice CT, angiography, intravascular ultrasound, derived morphology parameters (virtual histology, palpography, and echogenicity), and optical coherence tomography (OCT). FINDINGS: Clinical data were obtained from 29 of 30 patients. At 2 years, the device was safe with no cardiac deaths, ischaemia-driven target lesion revascularisations, or stent thromboses recorded, and only one myocardial infarction (non-Q wave). 18-month multislice CT (assessed in 25 patients) showed a mean diameter stenosis of 19% (SD 9). At 2-year angiography, the in-stent late loss of 0.48 mm (SD 0.28) and the diameter stenosis of 27% (11) did not differ from the findings at 6 months. The luminal area enlargement on OCT and intravascular ultrasound between 6 months and 2 years was due to a decrease in plaque size without change in vessel size. At 2 years, 34.5% of strut locations presented no discernible features by OCT, confirming decreases in echogenicity and in radiofrequency backscattering; the remaining apparent struts were fully apposed. Additionally, vasomotion occurred at the stented site and adjacent coronary artery in response to vasoactive agents. INTERPRETATION: At 2 years after implantation the stent was bioabsorbed, had vasomotion restored and restenosis prevented, and was clinically safe, suggesting freedom from late thrombosis. Late luminal enlargement due to plaque reduction without vessel remodelling needs confirmation. PMID- 19286090 TI - Percutaneous coronary interventions for non-acute coronary artery disease: a quantitative 20-year synopsis and a network meta-analysis. AB - BACKGROUND: Over the past 20 years, percutaneous transluminal balloon coronary angioplasty (PTCA), bare-metal stents (BMS), and drug-eluting stents (DES) succeeded each other as catheter-based treatments for coronary artery disease. We undertook a systematic overview of randomised trials comparing these interventions with each other and with medical therapy in patients with non-acute coronary artery disease. METHODS: We searched Medline for trials contrasting at least two of the four interventions (PTCA, BMS, DES, and medical therapy). Eligible outcomes were death, myocardial infarction, coronary artery bypass grafting, target lesion or vessel revascularisation, and any revascularisation. Random effects meta-analyses summarised head-to-head (direct) comparisons, and network meta-analyses integrated direct and indirect evidence. FINDINGS: 61 eligible trials (25 388 patients) investigated four of six possible comparisons between the four interventions; no trials directly compared DES with medical therapy or PTCA. In all direct or indirect comparisons, succeeding advancements in percutaneous coronary intervention did not produce detectable improvements in deaths or myocardial infarction. The risk ratio (RR) for indirect comparisons between DES and medical therapy was 0.96 (95% CI 0.60-1.52) for death and 1.15 (0.73-1.82) for myocardial infarction. By contrast, we recorded sequential significant reductions in target lesion or vessel revascularisation with BMS compared with PTCA (RR 0.68 [0-60.0.77]) and with DES compared with BMS (0.44 [0.35-0.56]). The RR for the indirect comparison between DES and PTCA for target lesion or vessel revascularisation was 0.30 (0.17-0.51). INTERPRETATION: Sequential innovations in the catheter-based treatment of non-acute coronary artery disease showed no evidence of an effect on death or myocardial infarction when compared with medical therapy. These results lend support to present recommendations to optimise medical therapy as an initial management strategy in patients with this disease. PMID- 19286093 TI - Heart failure. AB - Despite advances in management of heart failure, the condition remains a major public-health issue, with high prevalence, poor clinical outcomes, and large health-care costs. Risk factors are well known and, thus, preventive strategies should have a positive effect on disease burden. Treatment of established systolic chronic heart failure includes use of agents that block the renin angiotensin-aldosterone and sympathetic nervous systems to prevent adverse remodelling, to reduce symptoms and prolong survival. Diuretics are used to achieve and maintain euvolaemia. Devices have a key role in management of advanced heart failure and include cardiac resynchronisation in patients with evidence of cardiac dyssynchrony and implantation of a cardioverter defibrillator in individuals with low ejection fraction. Approaches for treatment of acute heart failure and heart failure with preserved ejection fraction are supported by little clinical evidence. Emerging strategies for heart failure management include individualisation of treatment, novel approaches to diagnosis and tracking of therapeutic response, pharmacological agents aimed at new targets, and cell-based and gene-based methods for cardiac regeneration. PMID- 19286094 TI - Slurred speech and spirochaetes. PMID- 19286091 TI - Safety and tolerability of SCH 530348 in patients undergoing non-urgent percutaneous coronary intervention: a randomised, double-blind, placebo controlled phase II study. AB - BACKGROUND: An antithrombotic drug is needed that safely reduces cardiovascular events in patients undergoing percutaneous coronary intervention (PCI). We therefore assessed the tolerability and safety of SCH 530348-an oral platelet protease-activated receptor-1 antagonist. METHODS: We randomly assigned patients aged 45 years or older and undergoing non-urgent PCI or coronary angiography with planned PCI to an oral loading dose of SCH 530348 (10 mg, 20 mg, or 40 mg) or matching placebo in a 3:1 ratio in a multicentre international study. Those in the SCH 530348 group who subsequently underwent PCI (primary PCI cohort) continued taking an oral maintenance dose (0.5 mg, 1.0 mg, or 2.5 mg per day), and patients in the placebo group continued placebo for 60 days. The primary endpoint was the incidence of clinically significant major or minor bleeding according to the thrombolysis in myocardial infarction (TIMI) scale. Both investigators and patients were unaware of treatment allocation. Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00132912. FINDINGS: 257 patients were assigned to placebo and 773 to SCH 530348. The primary endpoint occurred in 2 (2%) of 129, 3 (3%) of 120, and 7 (4%) of 173 patients, respectively, in the SCH 530348 10 mg, 20 mg, and 40 mg groups compared with 5 (3%) of 151 patients in the placebo group (p=0.5786). TIMI major plus minor bleeding occurred in 3 (2%) of 136, 5 (4%) of 139, and 4 (3%) of 138 patients given SCH 530348 0.5 mg, 1.0 mg, and 2.5 mg once per day, respectively (p=0.7561). INTERPRETATION: Oral SCH 530348 was generally well tolerated and did not cause increased TIMI bleeding, even when administered concomitantly with aspirin and clopidogrel. Further testing in phase III trials to accurately define the safety and efficacy of SCH 530348 is warranted. PMID- 19286092 TI - Cardiovascular prevention guidelines in daily practice: a comparison of EUROASPIRE I, II, and III surveys in eight European countries. AB - BACKGROUND: The first and second EUROASPIRE surveys showed high rates of modifiable cardiovascular risk factors in patients with coronary heart disease. The third EUROASPIRE survey was done in 2006-07 in 22 countries to see whether preventive cardiology had improved and if the Joint European Societies' recommendations on cardiovascular disease prevention are being followed in clinical practice. METHODS: EUROASPIRE I, II, and III were designed as cross sectional studies and included the same selected geographical areas and hospitals in the Czech Republic, Finland, France, Germany, Hungary, Italy, the Netherlands, and Slovenia. Consecutive patients (men and women /=30 kg/m(2)) increased from 25.0% in EUROASPIRE I, to 32.6% in II, and 38.0% in III (p=0.0006). The proportion of patients with raised blood pressure (>/=140/90 mm Hg in patients without diabetes or >/=130/80 mm Hg in patients with diabetes) was similar (58.1% in EUROASPIRE I, 58.3% in II, and 60.9% in III; p=0.49), whereas the proportion with raised total cholesterol (>/=4.5 mmol/L) decreased, from 94.5% in EUROASPIRE I to 76.7% in II, and 46.2% in III (p<0.0001). The frequency of self-reported diabetes mellitus increased, from 17.4%, to 20.1%, and 28.0% (p=0.004). INTERPRETATION: These time trends show a compelling need for more effective lifestyle management of patients with coronary heart disease. Despite a substantial increase in antihypertensive and lipid-lowering drugs, blood pressure management remained unchanged, and almost half of all patients remain above the recommended lipid targets. To salvage the acutely ischaemic myocardium without addressing the underlying causes of the disease is futile; we need to invest in prevention. PMID- 19286095 TI - Resource overkill: we can do more for less. PMID- 19286096 TI - House syndrome. PMID- 19286097 TI - A weight lifted. PMID- 19286099 TI - Problems of practice: a new outlet to share them & improve volunteer EMS. PMID- 19286100 TI - No bailout for EMS: running your system on less without running it into the ground. PMID- 19286101 TI - Barking back: how the pros control dogs. PMID- 19286102 TI - Broken heart syndrome: what a rare cardiomyopathy can teach us. PMID- 19286103 TI - The value quotient: looking at the combined effects of quality & cost. PMID- 19286104 TI - The Aussie anesthetic: why the U.S. should consider equipping medics with ketamine. PMID- 19286105 TI - When dead is only mostly dead: understanding which causes of sudden cardiac death have good outcomes. PMID- 19286106 TI - A lot of nerve: how to perform a full neurological assessment for medical & trauma patients. AB - Rational thought, control of bodily movement and interpretation of external stimuli comprise but a short list of activities controlled by the nervous system. Disease or injury can adversely affect this system, which in turn negatively affects the entire body. Given the critical importance of the nervous system, prehospital providers must be adept in assessing and treating emergencies involving this vital system. After a brief review of related anatomy and physiology, consider the cases that follow, which illustrate scenarios that call for a thorough and comprehensive neurological exam. PMID- 19286107 TI - The disappearing endotracheal tube: historic skill threatened by lack of practice & new devices. AB - When paramedics were first introduced in the 1970s, one of the most controversial aspects of their training program was endotracheal intubation (ETI). Prior to that, the skill of intubating the trachea was purely the domain of physicians and nurse anesthetists. Thus, it was difficult to secure time when paramedic students could practice their intubations skills on live patients. In actuality, many paramedics of that era were graduated without ever having the opportunity to perform an ETI on a living patient. PMID- 19286108 TI - Don't be a minimizer: avoiding the trap of underestimating signs & symptoms. PMID- 19286109 TI - Rig ragging. PMID- 19286110 TI - [The future of telemedicine in the management of sleep-related respiratory disorders]. PMID- 19286111 TI - [Analysis of the current situation and outlook of thoracic surgery]. AB - OBJECTIVE: The objective of this descriptive study was to analyze the current situation and forecast the future requirements for specialists in thoracic surgery, taking into account the number of doctors entering and those possibly leaving this specialty. MATERIAL AND METHODS: The data for this study were taken from the Spanish Society of Pulmonology and Thoracic Surgery (SEPAR) (n=304), Thoracic Surgeons' Club (n=122), and the Spanish Council of Medical Associations (n=225). We also took into account the current number of resident surgeons (n=84). Other specialists were included who are not recorded in these databases but who are known to be practicing (n=10). The total number of practicing specialists obtained was 211. RESULTS: There are currently 52 working thoracic surgery departments and the highest number of practicing specialists was recorded in Madrid (n=44), Catalonia (n=33), and Andalusia (n=33). The forecast number of retirements (at age 65 years) and incorporations of new specialists means that there will be a surplus of 57 thoracic surgeons in the next 5 years. CONCLUSIONS: Thoracic surgery needs to limit the intake of new trainee specialists for at least the next 5 years. PMID- 19286112 TI - [Bronchoalveolar lavage findings in patients with diffuse interstitial lung disease: prospective study of a cohort of 562 patients]. AB - OBJECTIVE: Study of the bronchoalveolar lavage (BAL) fluid in some interstitial lung diseases can reveal patterns typical to each disease and that can support the diagnosis. The objective of this study was to perform a descriptive analysis of the cytologic study and of the lymphocyte subpopulations in BAL fluid from patients with interstitial lung disease. MATERIAL AND METHODS: In this prospective, observational study of 562 patients between January 1991 and January 2005, BAL fluid was analyzed to determine the distribution of cell populations and of lymphocyte subsets: CD3, CD4, CD8, CD3(+)CD4(-)CD8(-), and CD56. RESULTS: The mean age was 53.4 years and 53.3% of the patients were women. The following diseases were studied: idiopathic pulmonary fibrosis (n=132), sarcoidosis (n=123), connective tissue diseases (n=133), cryptogenic organizing pneumonia (n=89), and extrinsic allergic alveolitis (n=85). Isolated lymphocytic alveolitis was common in sarcoidosis and extrinsic allergic alveolitis. Mixed alveolitis was the most common pattern in the other interstitial lung diseases. The CD4:CD8 ratio was the most useful parameter. It was high in sarcoidosis (median, 2.3); the ratio was low or inverted in the other interstitial lung diseases, with median values of 1.76 in idiopathic pulmonary fibrosis, 0.45 in extrinsic allergic alveolitis, 0.35 in cryptogenic organizing pneumonia, and 0.33 in the connective tissue diseases. CONCLUSIONS: BAL parameters, in association with clinical and radiologic data, help to discriminate between interstitial lung diseases. BAL should therefore be considered a very useful tool in clinical management, particularly when pulmonary biopsy is not conclusive or is not possible. PMID- 19286113 TI - [Trends in asthma mortality in Spain from 1960 to 2005]. AB - BACKGROUND AND OBJECTIVE: No data have been published on trends in asthma mortality in Spain between 1960 and 2005. The aim of this study was to trace these trends for both the general population and the 5- to 34-year age group in Spain. MATERIAL AND METHODS: The data on asthma mortality for the general population and by age and sex were obtained from the Spanish National Institute of Statistics and used to calculate the asthma mortality rates per 100 000 inhabitants. RESULTS: The overall asthma mortality rate per 100 000 inhabitants was 9.36 in 1960 and 2.22 in 2005, showing a reduction from 12.26 to 0.96 in males and from 6.58 to 3.44 in females. In the 5- to 34-year age group, the mortality rate for both sexes ranged from 0.4 to 0.1, showing great variability over time. CONCLUSIONS: Asthma mortality decreased in the period analyzed, but less in women than in men. The trend and rates in Spain are similar to those of most countries for which data are available. PMID- 19286114 TI - [Determining the alveolar component of nitric oxide in exhaled air: procedures and reference values for healthy persons]. AB - Nitric oxide (NO) production has been described using a 2-compartment model for the synthesis and movement of NO in both the alveoli and the airways. The alveolar concentration of NO (Ca(NO)), an indirect marker of the inflammatory state of the distal portions of the lung, can be deduced through exhalation at multiple flow rates. Our objective was to determine reference values for Ca(NO). The fraction of exhaled NO (Fe(NO)) was measured in 33 healthy individuals at a rate of 50mL/s; the subjects then exhaled at 10, 30, 100, and 200mL/s to calculate Ca(NO). A chemiluminescence analyzer (NIOX Aerocrine) was used to perform the measurements. The mean (SD) Fe(NO) was 15 (6)ppb. The mean Ca(NO) was 3.04 (1.30)ppb. These values of Ca(NO) measured in healthy individuals will allow us to analyze alveolar inflammatory behavior in respiratory and systemic processes. PMID- 19286115 TI - [Response to inhaled granulocyte-macrophage colony-stimulating factor in a patient with alveolar proteinosis]. AB - Pulmonary alveolar proteinosis is a rare disease characterized by the accumulation of lipoproteinaceous material derived from alveolar surfactant in the alveoli, with a consequent deterioration in gas exchange. Pathogenesis is related to impaired phagocytic function of alveolar macrophages. In recent years, a new treatment for pulmonary alveolar proteinosis-consisting of subcutaneous administration of granulocyte-macrophage colony-stimulating factor (GM-CSF)-has become available. The commonly accepted treatment, and the one to have shown greatest efficacy in pulmonary alveolar proteinosis, is whole lung lavage. Instead of subcutaneous administration, GM-CSF can also be inhaled as an aerosol. This route of administration of GM-CSF is safe and effective in the treatment of pulmonary alveolar proteinosis and represents an alternative to subcutaneous administration or whole lung lavage. We present a patient with pulmonary alveolar proteinosis who was treated with inhaled GM-CSF and describe her clinical and functional outcome after 1 year of treatment. PMID- 19286116 TI - [Choriocarcinoma of the lung]. AB - Primary choriocarcinoma of the lung is an extremely rare condition with a fatal outcome in most patients. We report the case of a 31-year-old woman with cough, chest pain, and hemoptysis, whose chest radiograph displayed nodules in the right upper lobe. After study and an unsuccessful attempt at histological characterization of the lesions, exploratory surgery was indicated. The patient underwent lobectomy and lymphadenectomy after diagnosis of a malignant process in the intraoperative biopsy. The definitive diagnosis was choriocarcinoma. Gonadal involvement was ruled out and the patient was referred to the oncology department for adjuvant therapy. She remains free of disease after 42 months. Although choriocarcinoma of the lung is extremely rare and it is hard to establish a standard treatment, in cases of localized disease, surgical resection followed by chemotherapy seems to be useful and can achieve long survivals. PMID- 19286117 TI - [Intrasternal parathyroid adenoma]. PMID- 19286118 TI - [Recurrent lithoptysis in a patient with bronchiectasis]. PMID- 19286120 TI - Nothing to lose: a harm reduction approach to routine HIV testing. PMID- 19286121 TI - Suggestions for strengthening voluntary counseling and testing services. PMID- 19286122 TI - Prevention with positives: a review of published research, 1998-2008. AB - HIV prevention education and counseling efforts have historically been directed toward those individuals considered at risk for exposure to HIV and assumed to be uninfected with HIV. In the late 1990s, prevention efforts began to include individuals who were HIV-infected. In 2003, the Centers for Disease Control and Prevention recommended that HIV prevention be incorporated into the medical care of persons living with HIV. This domain of HIV prevention work is known as prevention with positives or positive prevention, and research within this domain has been ongoing for a decade. This article provides a review of the scientific evidence within the prevention with positives domain from 1998 to 2008. A discussion is provided regarding early descriptive and formative studies as well as more recent and ongoing intervention trials specifically designed for persons living with HIV. A summary of current knowledge, a description of ongoing research, and gaps in knowledge are identified. Topics for future research are suggested. PMID- 19286123 TI - Effects of a culturally adapted HIV prevention intervention in Haitian youth. AB - This study assessed the impact of an 8-week community-based translation of Becoming a Responsible Teen (BART), an HIV intervention that has been shown to be effective in other at-risk adolescent populations. A sample of Haitian adolescents living in the Miami area was randomized to a general health education control group (n = 101) or the BART intervention (n = 145), which was based on the information-motivation-behavior (IMB) model. Improvement in various IMB components (i.e., attitudinal, knowledge, and behavioral skills variables) related to condom use was assessed 1 month after the intervention. Longitudinal structural equation models using a mixture of latent and measured multi-item variables indicated that the intervention significantly and positively impacted all IMB variables tested in the model. These BART intervention-linked changes reflected greater knowledge, greater intentions to use condoms in the future, higher safer sex self-efficacy, an improved attitude about condom use, and an enhanced ability to use condoms after the 8-week intervention. PMID- 19286124 TI - Life improvement, life satisfaction, and care arrangement among AIDS orphans in rural Henan, China. AB - The Chinese government's response to the increasing number of children orphaned in the HIV epidemic included setting up AIDS orphanages and supporting community based group homes for double orphans (children who lost both parents to HIV). The impact of these strategies, compared to traditional kinship care, on children's outcomes has not been studied in China. The purpose of this study was to compare perceived life improvement and life satisfaction among double orphans in three main care arrangements (group home, AIDS orphanage, kinship care) in two rural Chinese counties. Participants included 176 children from four orphanages, 30 from eight group homes, and 90 from kinship households. Results indicated that children living in government-supported group homes were more likely to report greater life improvement and positive attitudes toward their current lives than children in orphanages and kinship care. Results suggested that perceived life improvements may have resulted from access to basic needs in extremely poor communities. PMID- 19286125 TI - African American grandfamilies' attitudes and feelings about sexual communication: focus group results. AB - Little attention has been given to the responsibilities that African American grandparent caregivers face when they assume the role of raising their grandchildren. One particularly challenging task of grandparent caregivers is communicating with their grandchildren about sexuality issues. This study therefore examined the sexual communications and attitudes and feelings toward these communications between African American grandparent caregivers and their adolescent grandchildren (11-13 years of age). Focus group methodology was used to generate data about the grandparent/grandchild sexuality communication process and attitudes and feelings toward that process. The sample included 40 dyads of African American grandparents/grandchildren, for a total of 80 participants. Grandparents as well as their adolescent grandchildren desired open communication about sexuality issues, said that societal pressure makes sexual encounters hard to resist, and felt that they needed assistance with the sexuality communication process. Nurses can build upon the study's results to assist African American grandparents and their adolescent grandchildren with sexual communication. PMID- 19286126 TI - Self-help groups can improve utilization of postnatal care by HIV-infected mothers. AB - HIV prevention within maternal-child health services has increased in many developing countries, but many HIV-infected women in developing countries still receive insufficient postnatal care. This study explored the experience of 30 HIV infected women in Vietnam in accessing HIV-related postnatal care, the role of felt and enacted stigma in accessing services, and the effects of participation in a self-help group on utilization of available services. Many HIV-infected women were not provided with adequate information on postnatal care by health workers. Most women reported both felt and enacted stigma that affected their access to care. Involvement in self-help groups improved the women's self-esteem, increased knowledge about HIV, and had a positive effect on both felt and enacted stigma from family, community, and health services. These results suggest the need for better information provision and better referral systems within the health services and suggest that establishing self-help groups can diminish felt stigma and facilitate access to services for women and their children. PMID- 19286127 TI - ESVS guidelines. Invasive treatment for carotid stenosis: indications, techniques. AB - The European Society for Vascular Surgery brought together a group of experts in the field of carotid artery disease to produce updated guidelines for the invasive treatment of carotid disease. The recommendations were rated according to the level of evidence. Carotid endarterectomy (CEA) is recommended in symptomatic patients with >50% stenosis if the perioperative stroke/death rate is <6% [A], preferably within 2 weeks of the patient's last symptoms [A]. CEA is also recommended in asymptomatic men <75 years old with 70-99% stenosis if the perioperative stroke/death risk is <3% [A]. The benefit from CEA in asymptomatic women is significantly less than in men [A]. CEA should therefore be considered only in younger, fit women [A]. Carotid patch angioplasty is preferable to primary closure [A]. Aspirin at a dose of 75-325 mg daily and statins should be given before, during and following CEA. [A] Carotid artery stenting (CAS) should be performed only in high-risk for CEA patients, in high-volume centres with documented low peri-operative stroke and death rates or inside a randomized controlled trial [C]. CAS should be performed under dual antiplatelet treatment with aspirin and clopidogrel [A]. Carotid protection devices are probably of benefit [C]. PMID- 19286128 TI - Bacterial martyrdom: phagocytes disabled by type VI secretion after engulfing bacteria. AB - To colonize during disease and prevent consumption by environmental unicellular eukaryotes, bacteria often disrupt phagocytosis. In this issue, Ma et al. (2009) show that Vibrio cholerae delivers the actin-crosslinking T6SS effector VgrG-1 following phagocytosis. The effector then causes irreversible cytoskeleton destruction, leading to bystander protection of the extracellular bacterial population. PMID- 19286129 TI - Putting enterohemorrhagic E. coli on a pedestal. AB - Many pathogenic bacteria exploit host cytoskeletal pathways to promote infection. In this issue of Cell Host & Microbe, Weiss et al. (2009) identify the host factor IRSp53 as the missing link that connects two intracellular bacterial proteins, thereby completing an actin cytoskeletal signaling pathway critical to enterohemorrhagic Escherichia coli pathogenesis. PMID- 19286130 TI - This bud's for Vpu. AB - Successful viruses must overcome the body's immune defenses. In this issue of Cell Host & Microbe, Goffinet et al. (2009) provide evidence that the host protein CD317, the target of the HIV Vpu protein, is part of an ancient innate immune response directed against budding viruses. PMID- 19286131 TI - Taming tuberculosis: a challenge for science and society. AB - Despite scientific advances, we are losing ground against Mycobacterium tuberculosis, a pathogen with which a third of us share our lives. Challenges include fundamental biologic questions; hurdles for translational medicine; and societal deficits in resources, incentives, collaboration, and leadership. Fortunately, the crisis is inspiring scientific and organizational creativity. PMID- 19286132 TI - The Salmonella effector SptP dephosphorylates host AAA+ ATPase VCP to promote development of its intracellular replicative niche. AB - Virulence effectors delivered into intestinal epithelial cells by Salmonella trigger actin remodeling to direct pathogen internalization and intracellular replication in Salmonella-containing vacuoles (SCVs). One such effector, SptP, functions early during pathogen entry to deactivate Rho GTPases and reverse pathogen-induced cytoskeletal changes following uptake. SptP also harbors a C terminal protein tyrosine phosphatase (PTPase) domain with no clear host substrates. Investigating SptP's longevity in infected cells, we uncover a late function of SptP, showing that it associates with SCVs, and its PTPase activity increases pathogen replication. Direct SptP binding and specific dephosphorylation of the AAA+ ATPase valosin-containing protein (VCP/p97), a facilitator of cellular membrane fusion and protein degradation, enhanced pathogen replication in SCVs. VCP and its adaptors p47 and Ufd1 were necessary for generating Salmonella-induced filaments on SCVs, a membrane fusion event characteristic of the pathogen replicative phase. Thus, Salmonella regulates the biogenesis of an intracellular niche through SptP-mediated dephosphorylation of VCP. PMID- 19286134 TI - IRSp53 links the enterohemorrhagic E. coli effectors Tir and EspFU for actin pedestal formation. AB - Actin pedestal formation by pathogenic E. coli requires signaling by the bacterial intimin receptor Tir, which induces host cell actin polymerization mediated by N-WASP and the Arp2/3 complex. Whereas canonical enteropathogenic E. coli (EPEC) recruit these actin regulators through tyrosine kinase signaling cascades, enterohemorrhagic E. coli (EHEC) O157:H7 employ the bacterial effector EspF(U) (TccP), a potent N-WASP activator. Here, we show that IRSp53 family members, key regulators of membrane and actin dynamics, directly interact with both Tir and EspF(U). IRSp53 colocalizes with EspF(U) and N-WASP in actin pedestals. In addition, targeting of IRSp53 is independent of EspF(U) and N-WASP but requires Tir residues 454-463, previously shown to be essential for EspF(U) dependent actin assembly. Genetic and functional loss of IRSp53 abrogates actin assembly mediated by EHEC. Collectively, these data indentify IRSp53 family proteins as the missing host cell factors linking bacterial Tir and EspF(U) in EHEC pedestal formation. PMID- 19286133 TI - Translocation of a Vibrio cholerae type VI secretion effector requires bacterial endocytosis by host cells. AB - The type VI secretion system (T6SS) is a virulence mechanism common to several Gram-negative pathogens. In Vibrio cholerae, VgrG-1 is required for T6SS dependent secretion. VgrG-1 is also secreted by T6SS and displays a C-terminal actin crosslinking domain (ACD). Using a heterologous reporter enzyme in place of the ACD, we show that the effector and secretion functions of VgrG-1 are genetically dissociable with the ACD being dispensable for secretion but required for T6SS-dependent phenotypes. Furthermore, internalization of bacteria is required for ACD translocation into phagocytic target cells. Inhibiting bacterial uptake abolishes actin crosslinking, while improving intracellular survival enhances it. Otherwise resistant nonphagocytic cells become susceptible to T6SS mediated actin crosslinking when engineered to take up bacteria. Our results support a model for translocation of VgrG C-terminal effector domains into target cell cytosol by a process that requires trafficking of bacterial cells into an endocytic compartment where translocation is triggered by an unknown signal. PMID- 19286135 TI - Host cell entry by apicomplexa parasites requires actin polymerization in the host cell. AB - Apicomplexa are obligate intracellular parasites that actively invade host cells using their membrane-associated, actin-myosin motor. The current view is that host cell invasion by Apicomplexa requires the formation of a parasite-host cell junction, which has been termed the moving junction, but does not require the active participation of host actin. Using Toxoplasma gondii tachyzoites and Plasmodium berghei sporozoites, we show that host actin participates in parasite entry. Parasites induce the formation of a ring-shaped F-actin structure in the host cell at the parasite-cell junction, which remains stable during parasite entry. The Arp2/3 complex, an actin-nucleating factor, is recruited at the ring structure and is important for parasite entry. We propose that Apicomplexa invasion of host cells requires not only the parasite motor but also de novo polymerization of host actin at the entry site for anchoring the junction on which the parasite pulls to penetrate the host cell. PMID- 19286136 TI - Two mosquito LRR proteins function as complement control factors in the TEP1 mediated killing of Plasmodium. AB - Plasmodium development within Anopheles mosquitoes is a vulnerable step in the parasite transmission cycle, and targeting this step represents a promising strategy for malaria control. The thioester-containing complement-like protein TEP1 and two leucine-rich repeat (LRR) proteins, LRIM1 and APL1, have been identified as major mosquito factors that regulate parasite loads. Here, we show that LRIM1 and APL1 are required for binding of TEP1 to parasites. RNAi silencing of the LRR-encoding genes results in deposition of TEP1 on Anopheles tissues, thereby depleting TEP1 from circulation in the hemolymph and impeding its binding to Plasmodium. LRIM1 and APL1 not only stabilize circulating TEP1, they also stabilize each other prior to their interaction with TEP1. Our results indicate that three major antiparasitic factors in mosquitoes jointly function as a complement-like system in parasite killing, and they reveal a role for LRR proteins as complement control factors. PMID- 19286137 TI - HIV-1 antagonism of CD317 is species specific and involves Vpu-mediated proteasomal degradation of the restriction factor. AB - Mammals encode proteins that inhibit viral replication at the cellular level. In turn, certain viruses have evolved genes that can functionally counteract these intrinsic restrictions. Human CD317 (BST-2/HM1.24/tetherin) is a restriction factor that blocks release of human immunodeficiency virus type 1 (HIV-1) from the cell surface and can be overcome by HIV-1 Vpu. Here, we show that mouse and rat CD317 potently inhibit HIV-1 release but are resistant to Vpu. Interspecies chimeras reveal that the rodent-specific resistance and human-specific sensitivity to Vpu antagonism involve all three major structural domains of CD317. To promote virus release, Vpu depletes cellular pools of human CD317, but not of the rodent orthologs, by accelerating its degradation via the 20S proteasome. Thus, HIV-1 Vpu suppresses the expression of the CD317 antiviral factor in human cells, and the species-specific resistance to this suppression may guide the development of small animal models of HIV infection. PMID- 19286138 TI - A functional genomic screen identifies cellular cofactors of hepatitis C virus replication. AB - Hepatitis C virus (HCV) chronically infects 3% of the world's population, and complications from HCV are the leading indication for liver transplantation. Given the need for better anti-HCV therapies, one strategy is to identify and target cellular cofactors of the virus lifecycle. Using a genome-wide siRNA library, we identified 96 human genes that support HCV replication, with a significant number of them being involved in vesicle organization and biogenesis. Phosphatidylinositol 4-kinase PI4KA and multiple subunits of the COPI vesicle coat complex were among the genes identified. Consistent with this, pharmacologic inhibitors of COPI and PI4KA blocked HCV replication. Targeting hepcidin, a peptide critical for iron homeostasis, also affected HCV replication, which may explain the known dysregulation of iron homeostasis in HCV infection. The host cofactors for HCV replication identified in this study should serve as a useful resource in delineating new targets for anti-HCV therapies. PMID- 19286139 TI - Self awareness and the body image. AB - What mental representations give us the sense of our body as a unique object in the world? We investigated this issue in the context of the rubber hand illusion (RHI), an illusion of body image in which a prosthetic hand brushed synchronously, but not asynchronously, with one's own hand is perceived as actually being one's hand. We conducted a large-scale study of the RHI, and used psychometric analysis to reveal the structure of the subjective experience of embodiment [Longo et al. (2008). What is embodiment? A psychometric approach. Cognition,107, 978-998]. Here, we use this dataset to investigate the relation between incorporation of a rubber hand into the body image and the perceived similarity between the participant's hand and the rubber hand. Objective similarity (as measured by skin luminance, hand shape, and third-person similarity ratings) did not appear to influence participants' experience of the RHI. Conversely, incorporation of the rubber hand into the body image did affect the similarity that participants perceived between their own hand and the rubber hand. Participants who had experienced the RHI perceived their hand and the rubber hand as significantly more similar than participants who had not experienced the illusion. That is, embodiment leads to perceived similarity, but perceived similarity does not lead to embodiment. Furthermore, similarity ratings following the illusion were selectively correlated with some components of embodiment, but not with others. These results suggest an important role of a mental body image in the perception of the relation between the self and others. PMID- 19286140 TI - Long-term histologic response to synthetic and biologic graft materials implanted in the vagina and abdomen of a rabbit model. AB - OBJECTIVE: We sought to evaluate the effects of graft type, site of implantation, and ovariectomy on the long-term histologic response to graft materials used in pelvic reconstructive surgery. STUDY DESIGN: In all, 45 rabbits underwent ovariectomy or sham laparotomy and were implanted with polypropylene (PP) mesh (n = 23) or cross-linked porcine dermis (PS) (n = 22) in the vagina and abdomen. Grafts were harvested 9 months later and were processed for histology to evaluate the host inflammatory response and degree of tissue incorporation within the grafts. RESULTS: Polypropylene induced a milder (P < .007), more uniform response than PS, whereas PS elicited a more variable response and degraded by 9 months. Vaginal grafts had higher scores for inflammation (P = .005) and neovascularization (P < .001), and had lower scores for fibroblastic proliferation (P < .001) than abdominal grafts. Ovariectomy had no effect (P > .05). CONCLUSION: Synthetic and biologic grafts evoke different foreign body responses, which may have implications for surgical outcomes in women. PMID- 19286141 TI - Contractile properties of the denervated external anal sphincter. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of denervation on contractile properties of the external anal sphincter (EAS) of the female rat. STUDY DESIGN: Sham operation, pudendal nerve transection, pelvic neurectomy, or combined pudendal nerve transection/pelvic neurectomy was performed in young female rats. Contractile function of the EAS was determined after 2 weeks. RESULTS: Maximal force-generating capacity of the EAS was not impaired by bilateral pudendal denervation or pelvic neurectomy. Twitch tension, however, was decreased, and fatigability increased after pelvic neurectomy. Combined bilateral pudendal nerve-transection plus pelvic neurectomy resulted in compromised force generating capacity, decreased twitch tension, and increased fatigability of the anal sphincter. CONCLUSION: Subtle changes in EAS function are detectable after pelvic neurectomy, but not pudendal denervation. In contrast, combination pudendal and pelvic neurectomy resulted in severe compromise of EAS function. These data suggest that EAS function is relatively preserved unless injury occurs to > 1 source of innervation. PMID- 19286142 TI - Transient lipopolysaccharide-induced cytokine responses in the maternal serum and amniotic fluid of the guinea pig. AB - OBJECTIVE: The aim of this study was to characterize the pregnant guinea pig cytokine time course following a maternal inflammatory insult. STUDY DESIGN: Pregnant guinea pigs (n = 34) were injected intraperitoneally with 100 microg/kg lipopolysaccharide (LPS) at 70% gestation and euthanized at 24 hours, 48 hours, or 5 days. Control animals were euthanized at 70% gestation without LPS exposure. Interleukin (IL)-6, IL-1beta, and tumor necrosis factor-alpha (TNF-alpha) were quantified in the maternal serum and amniotic fluid by an enzyme-linked immunosorbent assay. RESULTS: IL-6 and IL-1beta concentrations were elevated in the maternal serum at 24 hours and returned to control levels by 5 days. In the amniotic fluid, IL-6 peaked at 48 hours and IL-1beta at 24 hours. TNF-alpha levels were not significantly increased. CONCLUSION: Maternal intraperitoneal LPS injection produces transient increases in cytokine concentrations in the maternal serum and amniotic fluid within 5 days, further implicating the cytokines as mediators of fetal white matter damage. PMID- 19286144 TI - Cookie cutter doesn't cut it. PMID- 19286143 TI - The impact of stress incontinence surgery on female sexual function. AB - OBJECTIVE: We sought to describe change in sexual function 2 years after surgery to treat stress urinary incontinence. STUDY DESIGN: This analysis included 655 women randomized to Burch colposuspension or sling surgery. Sexual activity was assessed by the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) among those sexually active at baseline and 2 years after surgery. RESULTS: Mean PISQ-12 total score improved from baseline 32.23 +/- 6.85 to 36.85 +/- 5.89. After surgery, fewer subjects reported incontinence (9% vs 53%; P < .0001), restriction of sexual activity as a result of fear of incontinence (10% vs 52%; P < .0001), avoidance of intercourse because of vaginal bulging (3% vs 24%; P < .0001), or negative emotional reactions during sex (9% vs 35%; P < .0001). Women with successful surgery had greater improvement PISQ-12 scores (5.77 vs 3.79; P < .006). Sexually active women were younger, thinner, and had lower Medical, Epidemiological, and Social Aspects of Aging scores (total and urge subscale) than sexually inactive women. CONCLUSION: Sexual function improves after successful surgery and does not differ between Burch and sling. PMID- 19286145 TI - Effect of experimental diabetes on the development and maintenance of vulvovaginal candidiasis in female rats. AB - OBJECTIVE: The purpose of this study was to develop an experimental model of diabetes in female rats and verify its influence on vulvovaginal candidiasis. STUDY DESIGN: The animals were divided into control and diabetic groups. Diabetes was induced with the use of an intravenous solution of alloxan (42 mg/kg bodyweight). One week after confirmation of hyperglycemia, the inoculation of Candida albicans yeast, previously standardized from a vaginal isolate, in concentrations of about 5 x 10(8), was performed. Infection control was made through vaginal culture, Papanicolaou cytology, and scanning electron microscopy (SCEM). RESULTS: The results pointed to different glycemias between the control (74.8 +/- 2.6) and experimental groups (543.1 +/- 12.1) and a significant bodyweight decrease (227.6 +/- 4.77 and 204 +/- 6.39, respectively). The positive infection was shown by culture, Papanicolaou test, and SCEM in the experimental group. CONCLUSION: Diabetes mellitus causes hyperglycemia, which was favorable to the vaginal colonization and infection by C albicans. PMID- 19286146 TI - Episiotomy use has changed. PMID- 19286147 TI - Elective appendectomy and cesarean delivery. PMID- 19286148 TI - To push or not to push? The patient had not disclosed a problem that would complicate labor. PMID- 19286149 TI - Differences and consequences in interobserver variation in intrapartum fetal heart rate assessment. PMID- 19286150 TI - Vulvar cancer in young women: demographic features and outcome evaluation. AB - OBJECTIVE: The objective of the study was to identify prognostic and environmental factors associated with vulvar carcinoma in young women. STUDY DESIGN: This study was a review of patients younger than 45 years who were diagnosed with vulvar squamous cell carcinoma between 1994 and 2006. RESULTS: Fifty-six patients were identified. Median age was 38 years and median follow-up was 25.3 months. Fifty-eight percent of patients presented with stage I disease; 77% smoked tobacco. Of patients with advanced disease, 53.3% were smokers, 40% had human papillomavirus (HPV) exposure, 46.7% had a history of vulvar intraepithelial neoplasia (VIN), and 6.7% were immunocompromised. Symptoms were present for more than 12 months in 47%, but symptom duration did not correlate with stage (P = .42) or positive lymph nodes (P = .28). Disease recurred in 10.7% and 5.4% died of disease. CONCLUSION: Young women with vulvar cancer tend to have early-stage disease, smoke, have a history of HPV, and have VIN. Many of the factors that place these patients at continuous risk are modifiable. PMID- 19286151 TI - Ethics and referral for abortion. PMID- 19286152 TI - The rate of retinal vessel dilation in severe retinopathy of prematurity requiring treatment. AB - PURPOSE: To assess whether the rate of change of retinal vessel diameter can help identify infants at the highest risk for severe retinopathy of prematurity (ROP). DESIGN: Thirty-five infants at risk for ROP were included in this prospective, longitudinal study. METHODS: Images were obtained using the NIDEK NM200D noncontact camera (NIDEK Inc, Aichi, Japan) at the time of ROP examinations in the intensive care unit. Vessel diameters were measured from digital fundus photographs of right eyes in a masked fashion using VesselMap image analysis software (IM-EDOS GmbH, Weimar, Germany). The rate of change of vessel diameter was calculated based on the linear regression slope and was compared between eyes in which type 1 ROP requiring treatment developed and in controls without ROP or with ROP less severe than type 1. RESULTS: Multivariate analysis showed that the group of eyes in which type 1 ROP developed had a greater increase in diameter over time in the inferior temporal veins (P = .01), superior temporal veins (P < .0001), mean temporal veins (P < .0001), superior temporal arteries (P = .02), and mean temporal arteries (P = .004). The area under receiver operator characteristic curve for venous diameter change was 0.96 for the superior temporal vein, 0.86 for the inferior temporal vein, and 0.96 for the mean temporal vein. CONCLUSIONS: On average, the rate of retinal vessel change was greater in eyes with type 1 ROP requiring treatment than in control eyes. The rate of venous diameter change had the best discriminative ability to differentiate between the 2 groups. PMID- 19286153 TI - Refractive lens exchange with foldable toric intraocular lens. AB - PURPOSE: To assess visual and refractive outcomes, and rotational stability after refractive lens exchange (RLE) with toric intraocular lens (IOL) implantation to correct ametropia and preexisting astigmatism. DESIGN: Prospective, nonrandomized, observational case series (self-controlled). METHODS: This prospective, nonrandomized, and self-controlled study included 32 eyes of 19 consecutive patients with more than 1.00 diopter (D) of preexisting corneal astigmatism having RLE with AcrySof Toric IOL implantation (Alcon Laboratories Inc, Fort Worth, Texas, USA). Uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), refractive sphere, and keratometric and refractive cylinder were recorded preoperatively and 6 months after surgery. Toric IOL axis shift was also measured. A patient satisfaction, visual phenomena, and spectacle dependency questionnaire was also carried out. RESULTS: At 6 months postoperatively, UCVA was 20/32 or better in 100% of the eyes, with 84.3% achieving 20/25 or better. One hundred percent of eyes achieved 20/25 or better BCVA. No eye lost >or=2 lines, 1 eye lost 1 line, 16 eyes did not change, 4 eyes gained 1 line, and 11 eyes gained >or=2 lines of BCVA after the surgery. Mean refractive cylinder was reduced significantly after surgery from -2.46 +/- 0.99 D to -0.53 +/- 0.30 D (P < .001). Vector analysis to compare attempted vs achieved correction showed that 100% of eyes were within +/- 1.00 D for the spherical equivalent, and 100% of eyes were within +/-0.50 D for the astigmatic components (J(0) and J(45)). Mean toric IOL axis rotation was 0.90 +/- 1.76 degrees, being 3GalNAc core in the glycan portion of the glycoprotein antigen from the parasite Echinococcus multilocularis have been accomplished. Trisaccharide Gal beta 1-->3(GlcNAc beta 1-->6)GalNAc alpha 1-OR (A), tetrasaccharide Gal beta 1-->3(Gal beta 1-->4GlcNAc beta 1-->6)GalNAc alpha 1-OR (D), and pentasaccharides Gal beta 1-->3(Gal beta 1-->4Gal beta 1-->4GlcNAc beta 1-->6)GalNAc alpha 1-OR (E) and Gal beta1-->3(Gal alpha 1-->4Gal beta 1-->4GlcNAc beta 1-->6)GalNAc alpha 1-OR (F) (R = 2-(trimethylsilyl)ethyl) were synthesized by block synthesis. The disaccharide 2-(trimethylsilyl)ethyl 2,3,4,6-tetra-O acetyl-beta-D-galactopyranosyl-(1-->3)-2-azido-4-O-benzyl-2-deoxy-alpha-d galactopyranoside served as a common glycosyl acceptor in the synthesis of the branched oligosaccharides. Moreover, linear trisaccharide Gal beta 1-->4Gal beta 1-->3GalNAc alpha 1-OR (B) and branched tetrasaccharide Gal beta 1-->4Gal beta 1- >3(GlcNAc beta 1-->6)GalNAc alpha 1-OR (C) were synthesized by stepwise condensation. PMID- 19286170 TI - [Single incision laparoscopic appendectomy (SILS): Initial experience]. PMID- 19286171 TI - [Colonic neoplasm with liver metastasis. Sequential treatment using laparoscopic surgery]. PMID- 19286172 TI - Hemispheric lateralization and language skill coherence in temporal lobe epilepsy. AB - Patients with an early onset of temporal lobe epilepsy (TLE) are at an increased risk for language reorganization. It is unknown whether this reorganization involves a full shift of all language skills to the contralateral hemisphere, or whether it can be partial and involve only a subset of language skills. In this study we report dominance concordance patterns for five separate language skills measured during the Intracarotid Amobarbital Procedure (IAP) for 124 TLE patients. We examined whether the language skills show similar or independent lateralization patterns. We compare these patterns in early versus late seizure onset groups with either a left or right temporal lobe seizure focus. The data showed that the rates of atypical representation ranged from 25.8% for reading to 14.5% of the sample for speech. A majority of patients (60%) showing atypical language representation do so on more than one skill. While multiple atypicalities were common, the proportion of patients showing atypical representation on all five skills was strikingly low (5.6% of the total sample). Our data suggest that language systems are not independent and do not shift and reorganize in isolation, and no pairs of skills seem more likely to reorganize than others. There was also evidence that language is not monolithic with all language skills reorganizing together. The latter suggests that the pressures compelling atypical representation may not work equally on all language skills. PMID- 19286173 TI - Association between thyroid hormone levels and 4,4'-DDE concentrations in pregnant women (Valencia, Spain). AB - INTRODUCTION: Several epidemiological studies have reported that some organochlorine compounds (OCs), such as polychlorobiphenyls (PCBs) and 1,1 dichloro-2,2-bis(4-chlorophenyl)ethylene (4,4'-DDE), may alter thyroid function. OBJECTIVE: The aim of the present study was to investigate the association of maternal serum OC concentrations of 4,4'-DDE and the sum of seven PCB congeners (PCB 28, 52, 101, 118, 138, 153, and 180) with thyroid hormone (TH) status. METHODS: We measured OC concentrations in 157 maternal serum samples at 12 weeks of pregnancy in a cohort from Valencia (Spain). Thyroid-stimulating hormone (TSH), total triiodothyronine (TT3), and free thyroxine (FT4) were measured as biomarkers of thyroid function in the same samples. Linear and logistic regression analyses were performed between OCs and TH levels, and variables were log transformed. RESULTS: Mothers with higher levels of 4,4'-DDE had higher odds of having TSH levels >2.5 mIU/L (OR=2.53; 95% CI=1.36; 4.73; p=0.004), and we found a significant negative association between serum 4,4'-DDE concentrations and FT4 levels (beta=-0.03; 95% CI=-0.05; 0.00; p=0.050) after adjustment for covariates and total lipids. No association was found between sum of PCBs and TH levels. CONCLUSION: Serum concentrations of 4,4'-DDE were associated with increased TSH and reduced FT4 but not TT3 levels. Our results suggest that some environmental chemicals may interfere with the thyroid system of pregnant women. The major role that maternal THs may play in fetal neurodevelopment makes these findings especially relevant. PMID- 19286174 TI - Site-specific effects of 17beta-estradiol in hornyhead turbot (Pleuronichthys verticalis) collected from a wastewater outfall and reference location. AB - Studies throughout the southern California bight have indicated persistent estrogenic activity in male hornyhead turbot (Pleuronichthys verticalis). Plasma 17beta-estradiol (E2) concentrations correlated with gonadal DNA damage in fish collected near a wastewater treatment plant outfall, but not from fish collected at the reference location. When the same species was collected from the same reference location and treated with E2, no relationship between uptake and gonadal DNA damage was observed. To evaluate the site-specific effects of E2 in fish from a wastewater outfall and fish from a reference location, male hornyhead turbot from each location were exposed to 15 microg/L aqueous E2 in a time-course experiment, with fish sampled every 12 h for 48 h. Concentrations of E2 were measured in the aqueous exposure and in plasma from the fish. Vitellogenin (vtg) was also measured in the plasma, and 8-oxo-7,8-dihydro-2'-deoxyguanosine levels in male gonads were measured as an indicator of DNA damage. Untreated fish from the outfall had significantly lower E2 in the plasma relative to the untreated reference fish, and this trend was consistent at each time point in the E2 treated fish. Vtg was significantly induced after 36 h of exposure in fish from both sites and no significant differences were observed between the sites. A significant increase of oxidative DNA damage was observed in E2-treated fish from the outfall population and the damage was significantly correlated with plasma E2 concentrations only in fish from the outfall after 48 h. These results indicated that there were significant differences in E2 disposition and gonadal genotoxicity between the hornyhead turbot populations following exposure to E2, suggesting that fish at wastewater outfalls may be more sensitive to DNA damage, which may be temporally related to concentrations of E2 in plasma. PMID- 19286175 TI - Human exposure to mercury in the vicinity of chlor-alkali plant. AB - The main objectives of our study were to estimate the impact of a mercury cell chlor-alkali (MCCA) complex in Rosignano Solvay (Tuscany, Italy) on the local environment and to assess mercury exposure of inhabitants living near the plant. Measurement campaigns of atmospheric Hg near the MCCA plant showed that the impact of the emitted Hg from the industry on the terrestrial environment is restricted to a close surrounding area. Total gaseous mercury concentrations in ambient air of inhabited area around the MCCA plant were in the range of 8.0-8.7 ng/m3 in summer and 2.8-4.2 ng/m3 in winter. Peaks of up to 100 ng/m3 were observed at particular meteorological conditions. Background levels of 2 ng/m3 were reached within a radius of 3 km from the plant. Reactive gaseous mercury emissions from the plant constituted around 4.2% of total gaseous mercury and total particulate mercury emission constituted around 1.0% of total gaseous mercury emitted. Analysis of local vegetables and soil samples showed relatively low concentrations of total mercury (30.1-2919 microgHg/kg DW in the soil; <0.05 111 microgHg/kg DW in vegetables) and methylmercury (0.02-3.88 microgHg/kg DW in the soil; 0.03-1.18 microgHg/kg DW in vegetables). Locally caught marine fish and fresh marine fish from the local market had concentrations of total Hg from 0.049 to 2.48 microgHg/g FW, of which 37-100% were in the form of methylmercury. 19% of analysed fish exceeded 1.0 microgHg/g FW level, which is a limit set by the European Union law on Hg concentrations in edible marine species for tuna, swordfish and shark, while 39% of analysed fish exceeded the limit of 0.5 microgHg/g FW set for all other edible marine species. Risk assessment performed by calculating ratio of probable daily intake (PDI) and provisional tolerable daily intake (PTDI) for mercury species for various exposure pathways showed no risks to human health for elemental and inorganic mercury, except for some individuals with higher number of amalgam fillings, while PDI/PTDI ratio for methylmercury and total mercury exceeded the toxicologically tolerable value due to the potential consumption of contaminated marine fish. PMID- 19286176 TI - Randomized controlled study of EMR versus endoscopic submucosal dissection with a water-jet hybrid-knife of esophageal lesions in a porcine model. AB - BACKGROUND: Endoscopic submucosal dissection (ESD) seems to be more effective than EMR for curative resection of GI mucosal neoplasia. However, ESD is technically difficult, hazardous, and time consuming. New technologies may overcome these drawbacks. OBJECTIVE: Our purpose was to compare a new simplified ESD technique with conventional EMR. DESIGN: Prospective, randomized, and controlled trial in anesthetized pigs. METHODS: A total of 25 esophageal areas with a diameter of 20 mm were marked with coagulation points in 6 pigs under general anesthesia. These lesions were then randomized to either EMR by use of the cap technique or ESD. Submucosal injection of saline solution was used for both methods. ESD was performed with a hybrid-knife (ESDH), which allows cutting and coagulation as well as a needleless, tissue-selective hydrodissection through an axial water-jet channel with a preselected pressure by using a high-pressure water-jet system (ErbeJet 2). Intraoperative bleedings were treated with hemostatic forceps. MAIN OUTCOME MEASUREMENTS: A comparison of the rates of complete resection of the lesions, the number and the size of resected specimens, the complication rate, and the procedural duration. RESULTS: Thirteen lesions were randomized to EMR and 12 to ESDH. ESDH achieved complete resection significantly more frequently than EMR (10/12 vs 6/13; P = .05). All ESDH resections were performed as a single piece, whereas a mean (SD) of 2.5 +/- 0.9 resections were needed for EMR (P < .05). The mean (SD) areas of the specimen were 3.63 +/- 1.20 cm(2) in the ESDH group and 1.83 +/- 0.78 cm(2) in the EMR group (P < .0001). ESDH was performed with a larger amount of fluid for injection and hydrodissection compared with EMR (78.1 +/- 32.8 mL vs 20.9 +/- 7.6 mL; P < .001). The procedural duration was longer for ESDH than for EMR (28.2 +/- 11.9 minutes vs 12.2 +/- 4.9 minutes; P < .001). ESDH and EMR caused bleedings in 6 of 12 and 5 of 13 of the cases (P = .09), respectively. Hemostasis could be achieved in all cases. There was no perforation in either group. LIMITATIONS: Resection of artificial non-neoplastic lesions, small numbers, an animal trial. CONCLUSIONS: ESDH achieves complete resection of esophageal lesions significantly more frequently with a fewer number of specimens than EMR. ESDH is more time consuming, but the procedural duration seems to be shorter than conventional ESD because there is no need for exchange of devices for injection and cutting. The easy use of water-jet-assisted repeated injections of fluids may explain why ESDH was as safe as EMRC. PMID- 19286177 TI - Early detection of extrahepatic bile-duct carcinomas in the nonicteric stage by using MRCP followed by EUS. AB - BACKGROUND: Most extrahepatic bile-duct carcinomas are usually diagnosed when they are already in an advanced stage, which is the main reason for the poor prognosis of this tumor. OBJECTIVE: To examine the usefulness of MRCP followed by EUS in the early diagnosis of extrahepatic bile-duct carcinoma in the nonicteric stage. PATIENTS: This study included patients who were nonicteric, who had abnormal serum concentrations of alkaline phosphatase and gamma glutamyl transpeptidase, and whose common hepatic duct was more than 8 mm in diameter on abdominal US because of unknown reasons. DESIGN: A single-center, prospective study. SETTING: An academic medical center. MAIN OUTCOME MEASUREMENTS: The sensitivity and specificity of MRCP followed by EUS for the early diagnosis of extrahepatic bile duct carcinoma in the nonicteric stage. RESULTS: A total of 142 patients who were nonicteric underwent prospective MRCP, and 26 of them underwent EUS. Ten patients (5 with stricture, 4 with filling defect, and 1 with no stricture or filling defect) had extrahepatic bile-duct carcinoma, including 5 patients with an incidentally diagnosed T1 stage tumor. The sensitivity and specificity of MRCP followed by EUS were 90% and 98%, respectively. LIMITATIONS: A single center and small number of patients. CONCLUSIONS: MRCP followed by EUS was highly sensitive and specific for the early diagnosis of extrahepatic bile duct carcinoma in the nonicteric stage, including T1 stage tumors. Filling defects, as well as stenosis in the bile duct, are important MRCP findings of T1 stage carcinoma. PMID- 19286178 TI - Risk factors for complications after ERCP: a multivariate analysis of 11,497 procedures over 12 years. AB - BACKGROUND: Complications of ERCP are an important concern. We sought to determine predictors of post-ERCP complications at our institution. METHODS: GI TRAC is a comprehensive data set of patients who underwent ERCP at our institution from 1994 through 2006. Logistic regression models were used to evaluate 4 categories of complications: (1) overall complications, (2) pancreatitis, (3) bleeding, and (4) severe or fatal complications. Independent predictors of complications were determined with multivariable logistic regression. RESULTS: A total of 11,497 ERCP procedures were analyzed. There were 462 complications (4.0%), 42 of which were severe (0.36%) and 7 were fatal (0.06%). Specific complications of pancreatitis (2.6%) and bleeding (0.3%) were identified. Overall complications were statistically more likely among individuals with suspected sphincter of Oddi dysfunction (SOD) (odds ratio [OR] 1.91) and after a biliary sphincterotomy (OR 1.32). Subjects with a history of acute or chronic pancreatitis (OR 0.78) or who received a temporary small-caliber pancreatic stent (OR 0.69) had fewer complications. Post-ERCP pancreatitis was more likely to occur after a pancreatogram via the major papilla (OR 1.70) or minor papilla (OR 1.54) and among subjects with suspected SOD with stent placement (OR 1.45) or without stent placement (OR 1.84). Individuals undergoing biliary-stent exchange had less-frequent pancreatitis (OR 0.38). Biliary sphincterotomy was associated with bleeding (OR 4.71). Severe or fatal complications were associated with severe (OR 2.38) and incapacitating (OR 7.65) systemic disease, obesity (OR 5.18), known or suspected bile-duct stones (OR 4.08), pancreatic manometry (OR 3.57), and complex (grade 3) procedures (OR 2.86). CONCLUSIONS: This study characterizes a large series of ERCP procedures from a single institution and outlines the incidence and predictors of complications. PMID- 19286179 TI - From iatrogenic digestive perforation to complete anastomotic disunion: endoscopic stenting as a new concept of "stent-guided regeneration and re epithelialization". AB - BACKGROUND: Iatrogenic intestinal tract perforation and anastomotic disunion traditionally required surgical treatment. Complete anastomotic break was considered until now as an absolute contraindication for endoscopic management. OBJECTIVE: The aim of this series was to show that endoscopic management is able to treat a spectrum of bowel wall breaks, from focal perforation to complete anastomotic disunion. SETTING: A single-center prospective cohort study. PATIENTS: Nine consecutive patients with nonmalignant gastrointestinal perforations were treated with endoscopic stenting between 2005 and 2008. Perforations were related to endoscopic perforations (4 cases: 2 esophageal and 2 colorectal), postoperative fistula or leakage (2 cases: 1 colorectal anastomosis and 1 esophageal), and complete anastomotic disunion (3 cases: 2 ileoanal anastomosis and 1 esophagogastric anastomosis). INTERVENTIONS: All 9 patients underwent endoscopic installation of fully covered stents under endoscopic and radiologic guidance, sometimes associated with simultaneous endoscopic collection drainage. Oral feeding was resumed when radiologic contrast studies showed no residual leak. RESULTS: The outcome in all 9 patients was favorable. Two migrated stents were replaced, and 2 stents were spontaneously expelled without consequence. All stents were withdrawn within an average of 5 weeks. LIMITATIONS: Uncontrolled pilot study, small sample size. CONCLUSION: The successful endoscopic management of bowel wall breaks ranging from perforation to complete postoperative disunion with fully covered stent could support a new concept of "stent-guided regeneration and re-epithelialization." Controlled trials are needed before this new endoscopic treatment can be proposed as a substitute for traditional treatments. PMID- 19286180 TI - Thirty years of overt, obscure GI bleeding solved by modern technology. PMID- 19286181 TI - Attempted removal and subsequent fragmentation of 3 self-expanding metal stents. PMID- 19286182 TI - Sleeve string capsule endoscopy for real-time viewing of the esophagus: a pilot study (with video). AB - BACKGROUND: Previous studies suggest that string capsule endoscopy is feasible, safe, accurate, highly acceptable, and likely to be proven as a more cost effective technique than conventional EGD. However, this technique needs proper high-level disinfection, and the string attachment is not easy to perform. We developed a modified esophageal capsule endoscopy, called sleeve string capsule endoscopy (SSCE). OBJECTIVE: To assess the feasibility and safety of SSCE in the evaluation of esophageal diseases. DESIGN: A pilot study. SETTING: Outpatient setting in a tertiary hospital, Shanghai, China. PATIENTS: Two healthy volunteers and 8 patients with suspected esophageal diseases were enrolled. INTERVENTIONS: The OMOM capsule endoscope was enclosed in a small, transparent, thin, latex sleeve, with a string attached to the caudal end. Both the sleeve and the string were for single use and were discarded after completion of the procedure. Two healthy volunteers without previous EGD and 8 patients who had undergone EGD swallowed the capsules. Pictures of the esophagus were viewed in real time. MAIN OUTCOME MEASUREMENTS: Discomfort associated with the procedure, quality, and diagnostic value of the pictures were documented. In addition, patient preference between SSCE and conventional EGD was recorded. RESULTS: SSCE was successfully carried out in the 10 subjects. The procedures were easy and safe to perform. No sleeves and strings were disrupted or broken, and no capsule was lost. The mean overall discomfort score during SSCE in the 8 patients was 2.88 (range 2-5). Pictures generated during SSCE were generally of high quality and produced identical diagnoses to those obtained by EGD in all 8 patients. In addition, all the 8 patients preferred SSCE to EGD. LIMITATIONS: This was a single-center nonrandomized study with a small sample size. CONCLUSIONS: SSCE was a feasible, easy-to-operate, and safe method for the diagnosis of esophageal diseases without the need for disinfection. SSCE appears to possess the same diagnostic capacity as that of EGD. PMID- 19286183 TI - Assessing the global availability of misoprostol. AB - OBJECTIVE: To assess the worldwide availability of misoprostol. Documenting the extent of misoprostol use in obstetrics-gynecology is difficult because the drug typically is unregistered for such indications. METHODS: Data for 2002-2007 on annual sales (measured in weight) to hospitals and retail pharmacies, plus manufacturer prices per 200-microg misoprostol, were analyzed for medications containing misoprostol alone or combined with a nonsteroidal anti-inflammatory drug (NSAID); regional and country-specific trends were identified. Consumer prices per pill are documented for all formulations of registered medications. RESULTS: Of the misoprostol sold worldwide, 70% was misoprostol-NSAID-combination drugs; of this, 91% was sold in North America and Western Europe. Asia sold the most misoprostol-only drugs; sales increased dramatically in Bangladesh (by 128%) and India (646%), where various low-price brands are sold. Misoprostol sales decreased in Latin America but increased in the Middle East-North Africa and Sub Saharan Africa; these regions generally had low amounts sold per population. CONCLUSION: Availability is improving in some low-income regions where misoprostol could significantly reduce maternal deaths due to postpartum hemorrhage and unsafe abortion. PMID- 19286184 TI - Simultaneous analysis of plasma thiols by high-performance liquid chromatography with fluorescence detection using a new probe, 1,3,5,7-tetramethyl-8-phenyl-(4 iodoacetamido)difluoroboradiaza-s-indacene. AB - The design, synthesis and properties of a new derivatizing reagent, 1,3,5,7 tetramethyl-8-phenyl-(4-iodoacetamido)difluoroboradiaza-s-indacene (TMPAB-I), for thiol groups are presented. Using the derivatization of TMPAB-I with thiols, a new high-performance liquid chromatographic method for measuring low-molecular weight thiol-containing compounds, including coenzyme A (CoA), glutathione, N acetylcysteine, cysteine, homocysteine (HCys) and 6-mercaptopurine has been developed. The reaction of TMPAB-I with thiols is specific, fast and stable for both TMPAB-I and the derivatives. A baseline separation of all the six derivatives is achieved by isocratic elution on reversed-phase column within 20 min with detection wavelengths of 500 and 510 nm for the excitation and emission, respectively, and the limits of detection (signal-to-noise ratio=3) are from 1.8 fmol (CoA) to 14.0 fmol (HCys), respectively, per 20 microL injection. The utility of the proposed method has been validated by measuring thiol-containing compounds in human plasma samples from healthy persons and patients with hypertension, with recoveries of 94.2-106.8%. PMID- 19286185 TI - Comprehensive supercritical fluid chromatography x reversed phase liquid chromatography for the analysis of the fatty acids in fish oil. AB - The separation of the phenacyl esters of the fatty acids originating from a fish oil extract by means of a comprehensive analysis using silver-ion (SI) supercritical fluid chromatography (SFC) and reversed phase liquid chromatography (RP-LC) in the first and second dimensions, respectively, is described. The combination ensured a high orthogonality and peak capacity, particularly when compared to the comprehensive RP-LC x 2RP-LC separation achieved by using a configuration with two columns in parallel in the second dimension. The construction of the SI-SFC x RP-LC interface consists of two two-position/ten port switching valves, of which one is equipped with two loops packed with octadecyl silica (ODS) particles. Compared to the SFC x RP-LC configuration described in an earlier publication, the peak capacity in the second dimension was increased. Water was not only added as make-up fluid to the SFC effluent to ensure analyte focusing, but also as rinsing medium of the loops prior to the transfer of the fractions to the second dimension. In the SFC dimension, high efficiency and loadability were obtained by coupling two wide-bore columns (4.6 mm ID) in series. Evaporative light scattering (ELSD) and ultraviolet (UV) detection with standard and high-pressure flow cells were evaluated in terms of data acquisition speed and suppression of signal interferences originating from the supercritical carbon dioxide (CO(2)) expansion. PMID- 19286186 TI - Rapid identification and quantification of methamphetamine and amphetamine in hair by gas chromatography/mass spectrometry coupled with micropulverized extraction, aqueous acetylation and microextraction by packed sorbent. AB - We developed a rapid identification and quantification method for the toxicological analysis of methamphetamine and amphetamine in human hair by gas chromatography/mass spectrometry coupled with a novel combination of micropulverized extraction, aqueous acetylation and microextraction by packed sorbent (MEPS) named MiAMi-GC/MS. A washed hair sample (1-5 mg) was micropulverized for 5 min in a 2 mL plastic tube with 250 microL of water. An anion-exchange sorbent was added to adsorb anionic interferences. After removing the residue with a membrane-filter unit, sodium carbonate and acetic anhydride was admixed in turn. Acetylation was completed in approximately 20 min at room temperature. The acetylated analytes in the reaction liquid were concentrated to an octadecylsilica sorbent packed in the needle of a syringe by a CombiPAL autosampler. Elution was carried out with 50 microL of methanol, and the entire eluate injected into a gas chromatograph using a programmable temperature vaporizing (PTV) technique. The time required for sample preparation and GC/MS analysis was approximately 1 h from a washed hair sample, and an evaporation process was not required. Ranges for quantification were 0.20-50 (ng/mg) each for methamphetamine and amphetamine using 1 mg of hair. Accuracy and relative standard deviation (RSD) were evaluated intraday and interday at three concentrations, and the results were within the limit of a guidance issued by U.S. Food and Drug Administration. For identification, full-scan mass spectra of methamphetamine and amphetamine were obtained using 5 mg of fortified hair samples at 0.2 ng/mg. The extraction device of MEPS was durable for at least 300 extractions, whereas the liner of the gas chromatograph should be replaced after 20-30 times use. The carry over was estimated to be about 1-2%. This sample preparation method coupled with GC/MS is fast and labor-saving in comparison with conventional methods. PMID- 19286187 TI - Enantioseparation of flurbiprofen on amylose-derived chiral stationary phase by supercritical fluid chromatography. AB - The separation of the enantiomers of flurbiprofen on an amylose-derived chiral stationary phase, Chiralpak AD-H, by supercritical fluid chromatography (SFC) under both linear and non-linear conditions is studied. Pulse injections were implemented using supercritical CO2 modified with methanol as a mobile phase at a temperature of 30 degrees C. At linear conditions, the isotherm is determined directly from the chromatogram. Under overload conditions, the elution profiles were described by competitive Langmuir and bi-Langmuir isotherm. Isotherm parameters were estimated using the inverse method and the effects of operation variables such as pressure and modifier composition were studied. The value of selectivity is from 1.9 to 2.1 while the value of resolution is from 5.3 to 11.8. The number of theoretical plates is always greater than 5000 indicating high efficiency of SFC. PMID- 19286188 TI - Thin-layer chromatography for the identification and semi-quantification of biogenic amines produced by bacteria. AB - The screening TLC method described enables the simultaneous identification and semi-quantification of eight biogenic amines with a large number of samples handled in a short period of time. The dansylation process time was drastically reduced from 80 to 18 min by increasing the reaction temperature to 100 degrees C. Bacteria could be classified as no, low, moderate or powerful amine producers when no spots were noticeable in TLC plates, less than 50 mg/L, from 50 to 500 mg/L, or more than 500 mg/L of amines were present in the decarboxylase broth medium, respectively. This TLC method is a simpler, faster and less expensive alternative to other methods, such as differential culture media, HPLC and even more sensitive than other TLC procedures. PMID- 19286189 TI - Determination of food emulsifiers in commercial additives and food products by liquid chromatography/atmospheric-pressure chemical ionisation mass spectrometry. AB - A new, reliable liquid chromatography/atmospheric-pressure chemical ionisation mass spectrometry (LC-APCI-MS) method was developed for the quantitative determination of food emulsifiers composed of mono- and diacylglycerols of fatty acids (E471 series) in complex food matrices. These additives are extremely interesting for the food industry because of their useful properties. Indeed, they improve the manufacture of products by acting as foams and creams stabilisers, crumb-softeners, or antistaling agents. The proposed method also allows us to qualitatively characterise new food emulsifiers composed of other acid esters of mono- and diacylglycerols (E472 series). The validation of the method was performed on blank minicake spiked samples for detection limits (reaching ppm levels), linearity, recovery, precision, and accuracy. The method was then successfully applied to commercial additives containing mixtures of emulsifiers, as well as to food products such as margarines and minicakes. PMID- 19286190 TI - Synthesis and morphology control of ZnO nanostructures in microemulsions. AB - ZnO nanostructures with different morphologies and optical properties were prepared by a simple microemulsion process, and PEG400 was used as a directing agent. The samples were characterized by TEM, XRD, FTIR, and TG-DTA analysis. The XRD spectra indicate that the ZnO crystal has a hexagonal wurtzite structure. Needle-like, columnar, and spherical ZnO samples were synthesized respectively with the increase of PEG400 concentration in Zn(NO(3))(2) solution. TEM images and thermogravimetric analysis reveal that the microemulsion interface and the PEG400 agent have a synergistic effect on the morphology and crystalline size transition of ZnO nanostructures. The optical properties of the samples were investigated by measuring the UV-Vis absorbance spectra at room temperature. All the samples exhibit strong UV absorption at around 365 nm. ZnO products with band gap energies at 3.06, 3.02, 2.95, and 2.85 eV were obtained with 0, 12.5, 25.0, and 50.0% of PEG400 in Zn(NO(3))(2) solution, respectively. The formation mechanism of the ZnO nanostructures was proposed and discussed in detail. The synergistic control of the microemulsion interface and the agent on the growth of crystal nuclei reported here provides an alternative approach for preparation of other well-defined nanostructures. PMID- 19286191 TI - Expression and clinical significance of CD147 in genitourinary carcinomas. AB - BACKGROUND: CD147/extracellular matrix metalloproteinase inducer (EMMPRIN) expressed by tumor cells stimulates peri-tumorous fibroblasts to produce matrix metalloproteinases (MMPs), thus contributing to tumor invasion and metastasis. To assess its suitability as a potential therapeutic target, as well as its association with the clinicopathologic features and the prognosis of patients, the expression of CD147/EMMPRIN in neoplastic tissues of the genitourinary system were analyzed. METHODS: CD147/EMMPRIN expression in 52 patients with renal carcinoma, 58 patients with bladder carcinoma, 101 patients with prostate carcinoma, 17 patients of penis carcinoma, and 17 patients of testis carcinoma were examined by immunostaining on paraffin-embedded tumor specimens using monoclonal antibodies. Then, the association of its expression with clinicopathologic characteristics to the patients' prognosis was analyzed. The RNA interference approach was used to silence CD147/EMMPRIN expression in the human prostate carcinoma cell line LNCAP and human bladder carcinoma cell line J82. The in vitro proliferative ability of CD147/EMMPRIN-deficient cells was determined by a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay. RESULTS: CD147/EMMPRIN was expressed in neoplastic tissues, but not in normal tissues. Positive expression was shown in 42 of 52 (80.77%) of the patients with renal carcinoma, 41 of 58 (70.69%) of the patients with bladder carcinoma, 67 of 101 (66.34%) of the patients with prostate carcinoma, 16 of 17 (94.12%) of the patients with penis carcinoma and testis carcinoma. Positive CD147/EMMPRIN staining was significantly associated with TNM stages and histological subtypes of patients with various urinary carcinomas (P < 0.05). In all five groups, for different expression levels of CD147/EMMPRIN, the patients with a highly positive expression of CD147/EMMPRIN had the poorest prognosis. The siRNA-treated cells exhibited significantly decreased growth ability compared with control cells in vitro. CONCLUSION: These results may assist in defining the suitability of CD147/EMMPRIN as a therapeutic target and as a method for predicting a poor outcome in patients with various urinary carcinomas. PMID- 19286192 TI - Efficacy and tolerability of Ginkgo biloba extract EGb 761 by type of dementia: analyses of a randomised controlled trial. AB - Secondary analyses of a randomised controlled trial were performed to find out whether treatment effects of Ginkgo biloba extract EGb 761 differed by type of dementia. Three hundred ninety-five patients aged 50 years or above, with dementia with neuropsychiatric features were treated with EGb 761 (240 mg/day) or placebo for 22 weeks. Patients scored between 9 and 23 on the Short Syndrome Test (SKT), a cross-culturally validated cognitive test battery. Their total score on the Neuropsychiatric Inventory (NPI) was at least 5. Efficacy was assessed by the SKT test battery (primary outcome measure), the Verbal Fluency Test, the Clock Drawing Test, the NPI, the Hamilton Rating Scale for Depression (HAMD), and the Gottfries-Brane-Steen Scale (GBS). Applying standard research diagnostic criteria 214 patients were diagnosed with Alzheimer's disease (probable AD or possible AD with cerebrovascular disease) and 181 with probable vascular dementia (VaD). Under EGb 761 treatment the SKT total score improved by -3.0+/-2.3 and -3.4+/-2.3 points in patients with AD and VaD, respectively, whereas the patients on placebo deteriorated by +1.2+/-2.5 and +1.5+/-2.2 points, respectively (p<0.01 for both drug-placebo differences). Significant drug-placebo differences were found for all secondary outcome variables with no major differences between AD and VaD subgroups. The rate of adverse events tended to be higher for the placebo group. PMID- 19286193 TI - Voxel-wise assessment of progression of regional brain atrophy in relapsing remitting multiple sclerosis. AB - OBJECTIVE: To perform voxel-wise assessments of regional brain atrophy state and rate in subjects with relapsing-remitting (RR) multiple sclerosis (MS). BACKGROUND: Recently, attention has focused on defining the tissue compartments and regions within which brain atrophy occurs. These regional measures of brain volume changes may help to better define the nature of the pathology underlying MS. In this context, specific regional measures of grey matter (GM) volume changes can be obtained by using the voxel-based morphometry (VBM) approach. METHODS: Fifty-nine subjects with RR MS underwent conventional MRI at baseline and after a mean follow-up period of 3 years. Cross-sectionally, two VBM analyses (SPM-VBM, based on the Statistical Parametric Mapping software package, and FSL VBM, based on the FMRIB Software Library tools) were performed to assess cortical GM volumes in RR MS patients compared to 25 age- and sex-matched normal controls (NC). Longitudinally, FSL-VBM and the regional extension of the SIENA method (SIENAr) were both used to assess regional brain atrophy rate in the RR MS patients and its relationship with increases in T(2)-weighted white matter (WM) lesion volume over the follow-up period. RESULTS: Widespread decrease in cortical GM volume was found in the RR MS patients compared to NC. Both SPM-VBM and FSL VBM showed similar involvement of cortical regions (frontal, temporal, parietal, occipital lobes and insula), with a close correlation between the numbers of significant voxels obtained with the two different procedures (r=0.73, p<0.001). After 3-year follow-up, both FSL-VBM and SIENAr showed a further significant reduction in GM volume in the lateral frontal and parietal cortices, bilaterally. Regional volume changes also appeared significantly pronounced in correspondence to the increase in T(2)-weighted WM lesion volume over the follow-up period. CONCLUSIONS: By using different methodologies, we showed similar widespread tissue loss in the cerebral cortex of patients with RR MS. This brain tissue loss further progresses over time, particularly in the fronto-parietal cortex and seems to be partially dependent upon the increase of lesion load. PMID- 19286194 TI - Can beta stiffness index be proposed as risk factor for dementia. AB - BACKGROUND AND AIM: Changes of arterial stiffness indicate alteration in arterial mechanics predisposing to the evolution of stroke or vascular dementia. The aim of this pilot study was to investigate whether ultrasound parameters, particularly beta stiffness index (BSI), should be further explored as independent markers or risk factors for dementia. PATIENTS AND METHODS: There were 38 demented patients included in this study (72.53+/-7.87 yrs) and 33 clinically healthy controls (68.85+/-3.52 yrs). Risk factors were noted and ultrasound measurements performed on common carotid artery (CCA) using eTracking software on Aloka ProSound ALPHA 10 with 13 MHz linear probe. Level of significance was p<0.05. RESULTS: Arterial hypertension was present in 24 patients, atrial fibrillation in 9, diabetes in 3, and hypercholesterolemia in 18. Hypertension was present as a single risk factor in 15 controls with average diastolic blood pressure significantly lower in patients. Significantly higher in patients were mean intima-media thickness, systolic and diastolic CCA diameters (CD), and mean BSI in CCA bilaterally. Linear regression analysis for groups of Alzheimer's dementia and vascular dementia proved that MMSE of the two groups relates to CCA diameter change (CDc) and BSI change explaining up to 5% of variability. CONCLUSIONS: CD, CDc and BSI should be monitored in patients with cognitive decline and further explored as possible independent markers or risk factors; in future studies groups of demented and non-demented patients should be age, sex and risk factor matched. PMID- 19286195 TI - Evaluation of the effect of thyrotropin releasing hormone (TRH) on regional cerebral blood flow in spinocerebellar degeneration using 3DSRT. AB - Thyrotropin releasing hormone (TRH) therapy improves cerebellar ataxia in patients with spinocerebellar degeneration (SCD). We investigated the effect of TRH on regional cerebral blood flow (rCBF) using the fully automated region of interest (ROI) technique, 3DSRT. Ten patients with SCD received TRH intravenously (2 mg/day) for 14 days and underwent brain perfusion single photon emission computed tomography before and after therapy. Clinical efficacy was assessed using the International Cooperative Ataxia Rating Scale (ICARS). The rCBF in each ROI was measured using the noninvasive Patlak plot method and calculated using 3DSRT. TRH significantly improved the ICARS scores and increased rCBF in the callosomarginal segment and cerebellum. Cerebellar rCBF increased in 4 of 5 patients with improved ICARS scores and in 3 of 5 patients without improved ICARS scores after TRH therapy. The correlation between the change in cerebellar rCBF and the improved ICARS score, however, was not significant. These findings indicate that TRH therapy may increase cerebellar rCBF in some patients with cerebellar forms of SCD and that 3DSRT may be useful for evaluating the efficacy of TRH for increasing CBF. The beneficial effects of TRH may be due to increased cerebellar rCBF or the increased rCBF may be a secondary effect of TRH therapy. PMID- 19286197 TI - Exogenous estrogen and urinary incontinence. PMID- 19286198 TI - Surgical implications of the obesity epidemic. PMID- 19286199 TI - Increased startle responses in interstitial cystitis: evidence for central hyperresponsiveness to visceral related threat. AB - PURPOSE: Hypersensitivity to visceral stimuli in interstitial cystitis/painful bladder syndrome may result from enhanced responsiveness of affective circuits (including the amygdala complex) and associated central pain amplification. Potentiation of the eyeblink startle reflex under threat is mediated by output from the amygdala complex and, therefore, represents a noninvasive marker to study group differences in responsiveness in this brain circuit. MATERIALS AND METHODS: Acoustic startle responses were examined in female patients with interstitial cystitis/painful bladder syndrome (13) and healthy controls (16) during context threat (application of muscle stimulation electrodes to the lower abdomen overlying the bladder), and cued conditions for safety (no stimulation possible), anticipation and imminent threat of aversive abdominal stimulation over the bladder. RESULTS: Patients showed significantly greater startle responses during nonimminent threat conditions (baseline, safe and anticipation periods) while both groups showed similar robust startle potentiation during the imminent threat condition. Higher rates of anxiety and depression symptoms in the patient group did not account for the group differences in startle reflex magnitude. CONCLUSIONS: Compared to controls, female patients with interstitial cystitis/painful bladder syndrome showed increased activation of a defensive emotional circuit in the context of a threat of abdominal pain. This pattern is similar to that previously reported in patients with anxiety disorders as well as those with irritable bowel syndrome. Since these circuits have an important role in central pain amplification related to affective and cognitive processes, these results support the hypothesis that the observed abnormality may be involved in the enhanced perception of bladder signals associated with interstitial cystitis/painful bladder syndrome. PMID- 19286200 TI - Does transrectal ultrasound probe configuration really matter? End fire versus side fire probe prostate cancer detection rates. AB - PURPOSE: We compared prostate cancer detection rates for the 2 most commonly used transrectal ultrasound prostate biopsy probes, end fire and side fire, to determine whether the probe configuration affects detection rates. MATERIALS AND METHODS: We evaluated 2,674 patients who underwent initial prostate biopsy between 2000 and 2008 with respect to prostate specific antigen, biopsy technique and pathological findings. Patients were divided into 1,124 in whom biopsies were performed with an end fire probe and 1,550 in whom biopsies were performed with a side fire probe. RESULTS: There was a significant difference in the overall cancer detection rate in the end vs side fire arms (45.8% vs 38.5%, p <0.001). In the subsets of patients with prostate specific antigen greater than 4 to 10 ng/ml or less and greater than 10 ng/ml a significant difference persisted (46.4% vs 38.9% and 61.7% vs 49.1%, p <0.004 and <0.015, respectively). There was also a significant difference in detection rates between probes in those who underwent 8 to 19 biopsy cores (p <0.009). Biopsies of greater than 20 cores failed to attain statistical significance (p >0.105). We also found that prostate volume, patient age, prostate specific antigen and hypoechoic findings were independent variables for predicting cancer detection on multivariate analysis (p <0.001). CONCLUSIONS: The type of probe significantly affects the overall prostate cancer detection rate, particularly in patients with prostate specific antigen greater than 4 ng/ml and/or nonsaturation (8 to 19 cores) prostate biopsy. This may be because the end fire probe allows better mechanical sampling of the lateral and apical regions of the peripheral zone, where cancer is most likely to reside. We set the stage for a randomized, controlled trial to confirm our observations. PMID- 19286201 TI - Renal sinus fat invasion in pT3a clear cell renal cell carcinoma affects outcomes of patients without nodal involvement or distant metastases. AB - PURPOSE: According to the 2002 American Joint Committee on Cancer TNM classification, perinephric and renal sinus fat invasion are classified as pT3a renal cell carcinoma. However, only a few studies have assessed the impact of sinus fat invasion on patient survival and with controversial results. We analyzed the impact of sinus fat invasion on cancer specific survival in a cohort of patients with pT3a clear cell renal cell carcinoma. MATERIALS AND METHODS: We retrospectively analyzed data on 115 consecutive patients treated with open radical nephrectomy for unilateral, sporadic pT3a clear cell renal cell carcinoma at our department from 1989 to 2006. All pathological specimens were rereviewed by a single uropathologist. The prognostic role of sinus fat invasion in cancer specific survival was assessed by Cox proportional hazards regression models. RESULTS: Ten patients had direct ipsilateral adrenal invasion and were excluded from analysis. A total of 105 patients with clear cell renal cell carcinoma were evaluated. Median followup was 38 months. In the overall population sinus fat invasion did not reach independent predictive status in terms of cancer specific survival on multivariate Cox regression analysis after adjusting for age, performance status, tumor dimension, tumor grade, synchronous metastases, nodal involvement, sarcomatoid differentiation and coagulative necrosis. In the subset of patients with pNx/pN0 M0 (83) the actuarial 5-year cancer specific survival was 71.9% and 45.5% for those with perinephric fat invasion only and sinus fat invasion, respectively (p = 0.025). Sinus fat invasion achieved an independent predictive role on multivariable Cox regression analysis (p = 0.048, HR 2.06). CONCLUSIONS: Sinus fat invasion in clear cell renal cell carcinoma significantly affects cancer specific survival in patients without nodal or distant metastases. However, sinus fat invasion is not associated with worse cancer specific survival in cases of metastatic disease. PMID- 19286205 TI - Yearly prostate specific antigen and digital rectal examination fluctuations in a screened population. AB - PURPOSE: Prostate biopsy is often recommended based on increases in prostate specific antigen and/or abnormal digital rectal examination. We investigated the stability of a single positive test during the next 3 consecutive years. MATERIALS AND METHODS: A total of 2,578 participants in a San Antonio screening cohort with 2 or more consecutive annual prostate specific antigen and digital rectal examination tests were identified. Occurrences of an increased prostate specific antigen (2.5 ng/ml or greater) followed by 1 or more nonincreased prostate specific antigen results were compared with similar fluctuations of digital rectal examination from abnormal to normal. RESULTS: In 2,272 men who did not have a biopsy during the study, in 23.3% of 744 incidences of an increased prostate specific antigen with 1 year of followup, the next prostate specific antigen was not increased. In 19.5% of 462 incidences of an increased prostate specific antigen with 2 years of followup, the next 2 consecutive prostate specific antigen levels were not increased. Finally, in 17.5% of 285 incidences of an increased prostate specific antigen with 3 years of followup, the next 3 consecutive prostate specific antigens were not increased. Rates were similar but lower in 221 men with 1 or more negative biopsies during the study and in 85 men in whom prostate cancer eventually developed during the study. In contrast, approximately 70% of abnormal digital rectal examinations were normal the following year even in patients with prostate cancer, and in the majority of incidences remained normal the next 2 to 3 consecutive years. CONCLUSIONS: Occurrences of reversed prostate specific antigen cut point or abnormal digital rectal examination based decisions to biopsy 1 or more years after the initial test are not uncommon, suggesting repetition of these tests. PMID- 19286207 TI - Hospice and emergency room use by disadvantaged men dying of prostate cancer. AB - PURPOSE: Hospice care has been found to improve symptom management, quality of death and quality of life at the end of life. We describe hospice use by a cohort of low income, uninsured men with prostate cancer enrolled in a public assistance program. We ascertained whether hospice enrollment was associated with a decrease in the number of prostate cancer related emergency room visits made before death. MATERIALS AND METHODS: We studied all 57 low income, uninsured men in a public assistance program who had died since its inception in 2001. The association between sociodemographic and clinical data, and hospice enrollment data were evaluated. RESULTS: The overall rate of hospice use was 28% (16 of 57 patients). The mean +/- SD duration of hospice enrollment before death was 44 +/- 43 days (median 34, range 2 to 143). Two patients (12%) were enrolled fewer than 7 days and none were enrolled more than 180 days. Prostate cancer related emergency room visits, adjuvant chemotherapy treatment, evidence of metastasis at initial presentation and death from prostate cancer were significantly associated with hospice use (p <0.05). We noted a trend toward fewer mean emergency room visits made by men enrolled in hospice care than by those not enrolled (0.7 +/- 1.3 vs 1.1 +/- 0.9, p = 0.15). CONCLUSIONS: Hospice use and the duration of enrollment by low income, uninsured men dying of prostate cancer was comparable to previously reported hospice use by insured individuals. Hospice enrollment was associated with fewer prostate cancer related emergency room visits. PMID- 19286209 TI - The case for neoadjuvant chemotherapy and cystectomy for muscle invasive bladder cancer. PMID- 19286210 TI - Association of nonsteroidal anti-inflammatory drugs, prostate specific antigen and prostate volume. AB - PURPOSE: Nonsteroidal anti-inflammatory drugs such as aspirin prevent cardiovascular disease and several prior studies suggest that nonsteroidal anti inflammatory drugs also decrease prostate inflammation and prostate cancer risk. We investigated the association between nonsteroidal anti-inflammatory drug use, prostate specific antigen and prostate volume, hypothesizing that there would be lower prostate specific antigen and prostate volume with nonsteroidal anti inflammatory drug use. MATERIALS AND METHODS: The Nashville Men's Health Study uses a multicenter, rapid recruitment protocol to collect clinical, biological, behavioral and body measurement data on 1,277 men older than 40 years who are scheduled for diagnostic prostate biopsy. Nonsteroidal anti-inflammatory drug use was ascertained by survey and clinical interview. Medical charts were reviewed to ascertain current prostate specific antigen, prostate volume and clinical diagnoses following biopsy. RESULTS: Approximately 46% of patients reported receiving nonsteroidal anti-inflammatory drugs, primarily aspirin (37%). After adjusting for age, race and other factors prostate volume was similar between aspirin users and nonusers (47.6 vs 46.0 ml, p = 0.16). In contrast, prostate specific antigen was significantly lower in aspirin users (7.3 vs 8.0 ng/ml, p = 0.01). The association between prostate specific antigen and aspirin was significant in men with latent prostate cancer (6.1 vs 7.3 ng/ml, p <0.01), marginal in patients with high grade prostatic intraepithelial neoplasia (5.0 vs 5.9 ng/ml, p = 0.09) and nonsignificant in those with a negative biopsy (5.6 vs 5.7 ng/ml, p = 0.64). The strongest prostate specific antigen-aspirin association was in men with cancer and a prostate volume of 60 ml or more (7.3 vs 12.7 ng/ml, p <0.01). CONCLUSIONS: Prostate specific antigen was significantly lower in aspirin users with latent cancer. Prostate volume was not associated with nonsteroidal anti-inflammatory drug use. Results suggest that aspirin may affect prostate cancer detection, suggesting a potential detection bias to address in future studies of nonsteroidal anti-inflammatory drugs and prostate cancer prevention. PMID- 19286211 TI - Prospectively packaged ilioinguinal lymphadenectomy for penile cancer: the disseminative pattern of lymph node metastasis. AB - PURPOSE: We prospectively evaluated the disseminative pattern of lymph node metastasis in penile cancer cases using packaged lymphadenectomy. In addition, we analyzed prognostic factors of the extent of lymph node metastasis. MATERIALS AND METHODS: Packaged inguinal lymphadenectomy was performed in 46 patients. A total of 24 patients with 1 or more positive inguinal lymph nodes underwent packaged iliac lymphadenectomy. Inguinal lymphadenectomy was divided into 3 packages, including medial inguinal, lateral inguinal and Cloquet's node packages. Medial and lateral inguinal packages were separated by the lateral surface of the femoral artery and the saphenous vein. Iliac lymphadenectomy was divided into 3 packages, including external iliac, obturator and common iliac packages. Clinicopathological features of the primary tumor and lymph nodes were correlated with the extent of lymph node metastasis. RESULTS: Of 92 groin basins 27 cases of inguinal lymphadenectomy and 7 of iliac lymphadenectomy had lymph node metastasis. Medial inguinal and external iliac packages were the most common involved regions in inguinal and iliac lymphadenectomy cases, respectively. No extended lymph node metastasis was observed in the absence of positive lymph nodes in the medial inguinal package. In groin basins with lymph node metastasis in the medial inguinal package extranodal extension was a significant predictor of extended lymph node metastasis. Cloquet's node was associated with iliac lymph node metastasis on univariate analysis. However, it was of limited predictive value in patients with 1 or 2 positive inguinal lymph nodes. CONCLUSIONS: The medial inguinal package defined in our study was the first involved lymph node region in penile cancer cases. Extranodal extension was an important predictor of extended lymph node metastasis beyond the medial inguinal package. PMID- 19286212 TI - Food for thought: obesity as the major contributing factor for most urological disorders. PMID- 19286213 TI - Long-term oncological outcomes in women undergoing radical cystectomy and orthotopic diversion for bladder cancer. AB - PURPOSE: We compared oncological outcomes in women undergoing radical cystectomy and orthotopic diversion for bladder transitional cell carcinoma. MATERIALS AND METHODS: From 1990 to 2005, 201 women underwent radical cystectomy, including 120 with an orthotopic neobladder. Median followup was 8.6 years. The clinical course, and pathological and oncological outcomes in these 120 women were analyzed and compared to those in 81 women undergoing radical cystectomy and cutaneous diversion during the same period. RESULTS: Overall 3 of 120 women (2.5%) who received a neobladder died perioperatively. In this group the tumor was pathologically organ confined in 73 patients (61%), extravesical in 18 (15%) and lymph node positive in 29 (24%). Overall 5 and 10-year recurrence-free survival was 62% and 55%, respectively. Five and 10-year recurrence-free survival in patients with organ confined and extravesical disease was similar at 75% and 67%, and 71% and 71%, respectively. Patients with lymph node positive disease had significantly worse 5 and 10-year recurrence-free survival (24% and 19%, respectively). One woman had recurrence in the urethra and 2 (1.7%) had local recurrence. As stratified by pathological subgroups, similar outcomes were observed when comparing women with an orthotopic neobladder to the 81 who underwent cutaneous diversion. CONCLUSIONS: Orthotopic diversion does not compromise the oncological outcome in women after radical cystectomy for bladder transitional cell carcinoma. Excellent local and urethral control may be expected. Women with node positive disease are at highest risk for recurrence. Similar outcomes were observed in women undergoing cutaneous diversion. PMID- 19286214 TI - Prostate cancer screening and treatment in the transplant population: current status and recommendations. AB - PURPOSE: We reviewed the current status of and recommendations for prostate cancer screening and treatment in the solid organ transplant population. MATERIALS AND METHODS: We performed a MEDLINE search to identify published data regarding prostate cancer screening, risk, treatment and outcomes in the solid organ transplant population. The literature was reviewed and summarized. RESULTS: Most data regarding outcomes of prostate cancer treatment in the transplant population are limited to case reports and small series, and primarily involve renal insufficiency. It does not appear that the development or natural history of prostate cancer is significantly affected by organ failure or subsequent transplantation. Thus, prostate specific antigen testing and screening protocols can be extrapolated from the general population. However, the balance of comorbid diseases and estimated limitations in life expectancy must be carefully considered, and emphasis should be placed on risk assessment. Prostatectomy appears to be feasible with outcomes comparable to those in the non-transplant population, while data regarding the use of radiation therapy are limited. CONCLUSIONS: The expansion of organ transplant criteria, including older donors and recipients, combined with improved allograft survival has enhanced the relevance of prostate cancer screening and treatment in this group. Greater awareness of the issues surrounding prostate cancer incidence, detection and natural history should promote improved data collection, screening and treatment of prostate cancer in the transplant population. PMID- 19286215 TI - New treatments for recurrent stress incontinence. PMID- 19286216 TI - The necessity of adrenalectomy at the time of radical nephrectomy: a systematic review. AB - PURPOSE: We describe the literature base pertaining to adrenalectomy at radical nephrectomy and present a pragmatic approach based on primary tumor and disease characteristics. MATERIALS AND METHODS: Literature searches were performed via the National Center for Biotechnology Information databases using various keywords. Articles that pertained to the concomitant use of adrenalectomy with radical nephrectomy were surveyed. RESULTS: The incidence of solitary, synchronous, ipsilateral adrenal involvement, ie that which is potentially curable with ipsilateral adrenalectomy along with nephrectomy, is much lower than previously thought at 1% to 5%. Evidence to date supports increased size and T stage, multifocality, upper pole location and venous thrombosis as risk factors for adrenal involvement. Cross-sectional imaging is now accurate at demonstrating the absence of adrenal involvement but still carries a significant risk of false positives. The morbidity of adrenalectomy is minimal except in those patients with metachronous contralateral adrenal metastasis in whom the impact of adrenal insufficiency can be devastating. Disease specific and overall survival of those undergoing radical nephrectomy, with or without adrenalectomy, are similar. The survival of patients with widespread metastatic disease is historically poor regardless of whether adrenalectomy is performed. There is evidence for a survival advantage in patients with isolated adrenal metastasis, although this group comprises no more than 2% of those undergoing surgery for renal tumors. CONCLUSIONS: The apparent benefit of ipsilateral adrenalectomy does not support it as a standard practice in all patients with normal imaging. However, it should be considered in select cases in which there are risk factors for adrenal involvement. PMID- 19286217 TI - Tumor size is associated with malignant potential in renal cell carcinoma cases. AB - PURPOSE: We evaluated our experience with renal cortical tumors to determine whether tumor size is associated with malignant histology and/or nuclear grade. MATERIALS AND METHODS: We identified 2,675 patients treated surgically at our institution for renal cell carcinoma or a benign tumor between 1989 and 2007. Histological subtype and tumor size were obtained from our kidney cancer database and logistic regression analysis was performed. RESULTS: Of the 2,675 tumors 311 (12%) were benign and 2,364 (88%) were renal cell carcinoma. The OR for the association of malignancy with tumor size was 1.16 (95% CI 1.11-1.22, p <0.001), indicating that each 1 cm increase in tumor size was associated with a 16% increase in the odds of malignancy. The incidence of benign tumors decreased from 38% for tumors less than 1 cm to 7% for tumors 7 cm or greater. In patients with clear cell renal cell carcinoma each 1 cm increase in tumor size increased the odds of high grade disease (Fuhrman grade 3-4) compared with low grade disease (Fuhrman grade 1-2) by 25% (OR 1.25, 95% CI 1.21-1.30, p <0.001). In this subset the incidence of high grade lesions increased from 0% for tumors less than 1 cm to 59% for tumors greater than 7 cm. CONCLUSIONS: Our results confirm previous observations suggesting that the risks of malignancy and high grade tumors increase with tumor size. Patients with small renal masses are at low risk for harboring a high grade clear cell malignancy, which may be useful during initial consultation. PMID- 19286220 TI - Predictors of oncological outcome after resection of locally recurrent renal cell carcinoma. AB - PURPOSE: Local renal cell carcinoma recurrence is rare after radical nephrectomy with curative intent. We examined our experience to describe the natural history of isolated local recurrence and characterize important prognostic factors that predict the outcome in patients treated with aggressive resection. MATERIALS AND METHODS: In an institutional database of 4,800 patients with renal cell carcinoma, of whom 2,945 underwent radical nephrectomy with curative intent, 54 were subsequently found to have isolated local recurrence in the renal fossa, ipsilateral adrenal gland or ipsilateral retroperitoneal lymph nodes, which was managed by surgical resection. In 69% of patients perioperative systemic therapy was done as an adjunct to surgical resection of local recurrence. RESULTS: Estimated median recurrence-free and cancer specific survival was 11 and 61 months, respectively. A positive surgical margin after resection of local recurrence, recurrent tumor size, sarcomatoid features in the recurrence specimen, abnormal serum alkaline phosphatase and increased lactate dehydrogenase at local recurrence were adverse risk factors associated with an increased risk of cancer specific death after resection for recurrent renal cell carcinoma. Patients with 0, 1 and greater than 1 adverse risk features demonstrated cancer specific survival times of 111, 40 and 8 months, respectively. CONCLUSIONS: Aggressive resection of isolated local recurrence offers durable local tumor control and cancer specific survival in a significant proportion of patients with renal cell carcinoma. Clinical and pathological prognostic features at local recurrence can be used for patient selection for surgery and also the thoughtful integration of systemic therapy with surgical extirpation. PMID- 19286222 TI - Sex cord stromal testicular tumors: a clinical series--uniformly stage I disease. AB - PURPOSE: Sex cord stromal testicular tumors are rare. Historically 10% of lesions are said to be malignant but to our knowledge there are no clinical or histological features that can accurately predict potential malignant behavior. Because of this, groups at some centers have advocated prophylactic retroperitoneal lymph node dissection in patients with clinical stage I disease. We reviewed our experience with these tumors to determine whether this policy is justified. MATERIALS AND METHODS: We retrospectively reviewed the records of all 38 men older than 18 years with sex cord stromal testicular tumors who were referred to the Wessex regional cancer center for treatment or pathological review during the 25-year period of 1982 to 2006. We then compared our series with a malignant sex cord stromal testicular tumor database generated from the world literature. RESULTS: All Wessex patients were treated with excision of the primary tumor alone and metastatic disease developed in none. All remained disease-free with an overall median survival of 6.8 years (range 1.4 to 25). Features in the literature favoring malignant behavior, ie metastatic disease, included larger tumors (mean 6.43 vs 1.71 cm), a high mitotic rate, tumor necrosis, angiolymphatic invasion, infiltrative margins and extratesticular extension (each p <0.0001). The malignant group had an overall median survival of 2.3 years (range 0.02 to 17.3). CONCLUSIONS: No patient had disease progression in our study, which is to our knowledge the largest reported United Kingdom series of sex cord stromal testicular tumors. Our data suggest that malignancy is uncommon and prophylactic retroperitoneal lymph node dissection is unjustified for clinical stage I disease. PMID- 19286223 TI - Learning to control cyclophosphamide induced cystitis. PMID- 19286224 TI - Fournier's Gangrene: population based epidemiology and outcomes. AB - PURPOSE: Case series have shown a Fournier's gangrene mortality rate of 20% to 40% with an incidence of as high as 88% in some studies. Because to our knowledge there are no population based data, we used a national database to investigate the epidemiology of Fournier's gangrene. MATERIALS AND METHODS: We used the State Inpatient Databases, the largest hospital based database available in the United States, which includes 100% of hospital discharges from participating states. Inpatients diagnosed with Fournier's gangrene (ICD-9 CM 608.83) who underwent genital/perineal debridement or died in the hospital were identified from 13 participating states in 2001 and from 21 in 2004. Population based incidence, regional trends and case fatality rates were estimated. RESULTS: We identified 1,641 males and 39 females with Fournier's gangrene. Cases represented less than 0.02% of hospital admissions. The overall incidence was 1.6/100,000 males, which peaked in males who were 50 to 79 years old (3.3/100,000) with the highest rate in the South (1.9/100,000). The overall case fatality rate was 7.5%. Patients with Fournier's gangrene were rarely treated at hospitals (mean +/- SD 0.6 +/- 1.2 per year, median 0, range 0 to 23). Overall 0 to 4 and 5 or greater cases were treated at 66%, 17%, 10%, 4%, 1% and 1% of hospitals, respectively. CONCLUSIONS: Patients with Fournier's gangrene are rarely treated at most hospitals. The population based mortality rate of 7.5% was substantially lower than that reported in case series from tertiary care centers. PMID- 19286225 TI - Metabolic complications of androgen deprivation therapy for prostate cancer. AB - PURPOSE: Androgen deprivation therapy has a variety of well recognized adverse effects including vasomotor flushing, loss of libido, fatigue, gynecomastia, anemia and osteoporosis. This review focuses on the more recently described metabolic complications of androgen deprivation therapy including obesity, insulin resistance and lipid alterations as well as the association of androgen deprivation therapy with diabetes and cardiovascular disease. MATERIALS AND METHODS: We reviewed the medical literature using the PubMed(R) search terms prostate cancer, androgen deprivation therapy, gonadotropin-releasing hormone agonists, obesity, insulin resistance, lipids, diabetes, cardiovascular disease and myocardial infarction. We provide a focused review and our perspective on the relevant literature. RESULTS: Androgen deprivation therapy decreases lean mass and increases fat mass. It also decreases insulin sensitivity while increasing low density lipoprotein cholesterol, high density lipoprotein cholesterol and triglycerides. Consistent with these adverse metabolic effects, androgen deprivation therapy may be associated with a greater incidence of diabetes and cardiovascular disease. Some of these androgen deprivation therapy related metabolic changes (obesity, insulin resistance and increased triglycerides) overlap with features of the metabolic syndrome. However, in contrast to the metabolic syndrome, androgen deprivation therapy increases subcutaneous fat and high density lipoprotein cholesterol. CONCLUSIONS: Androgen deprivation therapy increases obesity, decreases insulin sensitivity and adversely alters lipid profiles. It may be associated with a greater incidence of diabetes and cardiovascular disease. The benefits of androgen deprivation therapy should be weighed against these and other potential harms. Little is known about the optimal strategy to mitigate the adverse metabolic effects of androgen deprivation therapy. Thus, we recommend an emphasis on existing strategies for screening and treatment that have been documented to reduce the risk of diabetes and cardiovascular disease in the general population. PMID- 19286227 TI - Retroperitoneal lymph node dissection in patients with high risk testicular cancer. AB - PURPOSE: In patients with testicular cancer the percent of embryonal carcinoma and lymphovascular invasion in the primary tumor have been identified as risk factors for occult metastatic disease. We reviewed differences between primary and post-chemotherapy retroperitoneal lymph node dissection in patients at high risk. MATERIALS AND METHODS: Patients who underwent retroperitoneal lymph node dissection at our institution from 1993 to 2006 were identified and the clinical charts were reviewed. A total of 247 patients with orchiectomy specimens containing greater than 30% embryonal carcinoma were identified and perioperative data were obtained. RESULTS: Of 247 patients 133 (53%) had greater than 30% embryonal carcinoma, including 76 (57%) with combined lymphovascular invasion. Median followup was 3.49 years. Of the patients 76 (57%) and 57 (43%) underwent primary and post-chemotherapy retroperitoneal lymph node dissection, respectively, of whom most received bleomycin, etoposide and cisplatin. Positive lymph nodes were identified at surgery in 37 (49%) and 35 patients (61%) with primary and post-chemotherapy retroperitoneal lymph node dissection, respectively. Of patients with negative pathological findings at surgery surveillance computerized tomography postoperatively identified retroperitoneal masses in 2 (5%) and 3 (14%) of those who underwent a primary and a post chemotherapy procedure, respectively. Operative data on the primary vs post chemotherapy groups showed an estimated blood loss of 166 vs 371 cc, an operative time of 2.7 vs 3.3 hours and a hospital stay of 4.4 vs 4.7 days. There were no deaths in either group. CONCLUSIONS: Patients with greater than 30% embryonal carcinoma with or without lymphovascular invasion are at significant risk for metastatic disease and they can be successfully treated with primary retroperitoneal lymph node dissection. Recurrence rates based on computerized tomography evaluation were low and similar between the chemotherapy and nonchemotherapy treated groups. PMID- 19286230 TI - The use of delta(15)N in assessing sewage stress on coral reefs. AB - While coral reefs decline, scientists argue, and effective strategies to manage land-based pollution lag behind the extent of the problem. There is need for objective, cost-effective, assessment methods. The measurement of stable nitrogen isotope ratios, delta(15)N, in tissues of reef organisms shows promise as an indicator of sewage stress. The choice of target organism will depend upon study purpose, availability, and other considerations such as conservation. Algae are usually plentiful and have been shown faithfully to track sewage input. The organic matrix of bivalve shells can provide time series spanning, perhaps, decades. Gorgonians have been shown to track sewage, and can provide records potentially centuries-long. In areas where baseline data are lacking, which is almost everywhere, delta(15)N in gorgonians can provide information on status and trends. In coral tissue, delta(15)N combined with insoluble residue determination can provide information on both sewage and sediment stress in areas lacking baseline data. In the developed world, delta(15)N provides objective assessment in a field complicated by conflicting opinions. Sample handling and processing are simple and analysis costs are low. This is a method deserving widespread application. PMID- 19286231 TI - [Evaluation of the quick diagnosis and treatment program for lung cancer]. AB - OBJECTIVE: To analyze the efficiency of the program of quick diagnosis and treatment (PDTR, programa de diagnostico y tratamiento rapido) of lung cancer established in the Hospital de la Santa Creu i Sant Pau of Barcelona to review the epidemiology of lung cancer. METHODS AND MATERIALS: Fifty-eight patients with lung cancer were studied. Twenty-nine of them were included in the program between October 2005 and May 2006, and the remaining were randomly selected among those diagnosed the year before (control group). Time between first visit, diagnosis and treatment and other variables (age, sex, histological type and TNM stage) were compared between groups. RESULTS: Significant differences were found between the two groups. PDTR patients had a mean time between first visit and treatment of 26.7 days (Standard Deviation [SD]=13.6), whereas this was 84 days (SD=53) in the control group. The PDTR group had a lower TNM stage, but statistical significance was only found in N (lymph node involvement) (p=0.007). CONCLUSION: Most patients included in the PDTR program spend less than 30 days between first visit and treatment, which represents a significant reduction in time (p<0.001). The effect on prognosis is controversial and will need long term studies. PMID- 19286232 TI - [Radiologia and the circuit that fails to square.]. PMID- 19286233 TI - [Why should a radiologist publish?]. PMID- 19286234 TI - [Intracranial cavernous angioma.]. AB - We report the case of a 13-year-old boy who presented at the emergency department for nausea and vomiting with ataxia and dissymmetry. He had strabismus as a consequence of palsy of the VI cranial nerve when 9 months old that was attributed to an allergy to penicillin. He had no relevant family history. Urgent CT and posterior MRI examination showed multiple supratentorial and infratentorial cavernous angiomas, one of which was located in the brainstem and presented acute hemorrhage, causing the symptoms. Angiography performed later showed no evidence of vascular malformations. The patient was initially managed conservatively, but the cavernous angioma in the brainstem was surgically extirpated after the patient's condition progressively worsened. PMID- 19286235 TI - [Magnetic resonance imaging of the left ventricle in healthy subjects: normal values for morphology, function, perfusion, and viability]. AB - OBJECTIVE: To define normal values of MRI parameters related to cardiac morphology, function, perfusion, and delayed enhancement of the left ventricle and to analyze differences based on age and sex. MATERIAL AND METHODS: We used 1.5T and 3T MRI scanners to analyze 18 healthy subjects ranging in age from 15 to 77 years old. Dedicated image processing software (Cardio-MR, View Forum, Philips Medical Systems) was used to evaluate morphological and functional parameters (end-diastolic and end-systolic volume, stroke volume, ejection fraction, cardiac output, wall mass, wall thickness, wall thickening, wall motion), perfusion parameters (relative maximum upslope, relative maximum enhancement), and delayed enhancement (percentage of late hyperenhancement). Student's t-test was used for statistical analyses. RESULTS: Sex differences were observed: end-diastolic and end-systolic volumes, stroke volume, and wall mass were significantly higher in men and the ejection fraction was significantly larger in women. Healthy subjects over 45 years of age had significantly greater wall thickness. CONCLUSION: We report cardiac MRI reference values for morphological, functional, perfusion, and delayed enhancement parameters. Sex and age should be taken into account as important variables related to some of these parameters. PMID- 19286236 TI - [Abdominal manifestations in von Hippel Lindau disease in a group of 7 patients and literature review.]. AB - OBJECTIVE: To evaluate the radiologic findings (ultrasound, computed tomography and magnetic resonance) of the abdominal manifestations in von Hippel-Lindau disease (VHL). MATERIAL AND METHODS: We retrospectively reviewed the imaging studies performed to seven patients diagnosed of VHL in our hospital. RESULTS: In all exmined patients, abdominal involvement was found. All of them showed renal affectation: cysts or renal cell carcinoma. Pancreatic involvement was found in the majority of patients: simple cysts, serous cystadenomas and one neuroendocrine tumor. One patient had a papillary cystadenoma of the broad ligament. PMID- 19286237 TI - [Quantification of left ventricular function and mass in dual-source CT (DSCT).]. AB - OBJECTIVES: To evaluate the interobserver agreement in quantifying left ventricular function and mass and to assess the accuracy of conventional manual contour tracing compared to semiautomatic segmentation analysis software. MATERIAL AND METHODS: Twenty consecutive subjects who underwent cardiac DSCT with retrospective ECG-gating were included. Two different multiphase image reconstructions were done in 5% steps throughout the entire cardiac cycle (0-95% of the R-R interval) with effective slice thickness of 1mm in the axial plane and 8mm in the short-axis orientation. Left ventricular function and mass were calculated by two independent observers, tracing endocardial and epicardial borders manually and using a semiautomatic software tool (Circulation II, Siemens). Ejection fraction (EF), end-diastolic volume (EDV), end-systolic volume (ESV), stroke volume (SV), cardiac output (CO), and myocardial mass were evaluated by two independent observers blind to each other's assessments. The interobserver agreement and the reliability of the different segmentation methods were calculated. The time required for manual contouring and semiautomatic contour tracing was also registered. RESULTS: We found an excellent correlation (r>0.94; p<0.05) between the two independent observers for the quantification of left ventricular function and mass. Left ventricular functional parameters derived from semiautomatic contour software and conventional manual tracing method were not significantly different (p>0.05). The semiautomatic contour detection algorithm overestimated LV mass significantly compared with the manual contouring method (mean difference 29.45+/-1.64g; p<0.05). The time needed to calculate these parameters with the semiautomatic tool was significantly lower (248.85+/-99.8s) than with manual contouring (452.7+/-73.92s) (p<0.05). CONCLUSIONS: Interobserver agreement for quantifying left ventricular function and mass using DSCT is excellent. Despite overestimating left ventricular mass, the semiautomatic software tool allows cardiac parameters to be quantified with the same reliability as the conventional manual method in half the time. PMID- 19286238 TI - Scrub typhus in Darjeeling, India: opportunities for simple, practical prevention measures. AB - To identify risk factors for scrub typhus in Darjeeling, India, we compared 62 scrub typhus cases (acute fever with eschar and specific IgM) with 62 neighbourhood controls. Cases were more likely to live close to bushes [matched odds ratio (MOR) 10; 95% CI 2.3-63] and wood piles (MOR 3.5; 95% CI 1.5-9.5), to work on farms (MOR 10; 95% CI 2.7-63), to observe rodents at home (MOR 3.6; 95% CI 1.4-11) and at work (MOR 9; 95% CI 2.4-57), and to rear domestic animals (MOR 2.4; 95% CI 1.1-5.7). Cases were less likely to wash after work (MOR 0.4; 95% CI 0.1-0.9) and change clothes to sleep (MOR 0.2; 95% CI 0.1-0.5). A cleaner, rodent controlled environment may prevent exposure to scrub typhus. Personal protection measures and better hygiene could further reduce individual risk. PMID- 19286239 TI - Achieving trachoma control in Ghana after implementing the SAFE strategy. AB - The Ghana Health Service plans to eliminate blinding trachoma by 2010 and has implemented the SAFE strategy since 2001. The programme impact was assessed in all endemic districts. A two-stage, cluster random sample of 720 households was selected in each of 18 endemic districts in Upper West and Northern Regions. All eligible residents were examined for trachoma signs. Household environmental risk factors were assessed. In total, 74,225 persons from 12,679 households were examined. Prevalence of trachomatous inflammation-follicular in 1-9 year-old children was 0.84% (95% CI 0.63-1.05, range of point estimates by district 0.14 2.81%) and prevalence of trichiasis in adults aged > or = 15 years was 0.31% (95% CI 0.24-0.38, range by district 0.00-1.07%). An estimated 4950 persons have trichiasis, of whom 72.6% are aged > or = 60 years and 71.4% are women. Latrines were observed in 11.6% of households and 79.2% of interview respondents reported use of an improved water source. Active trachoma is no longer a public health problem in Ghana after successful implementation of the SAFE strategy. The programme should maintain health education, advocate for improved water and sanitation and focus on providing surgery. Surveillance activities are needed to ensure sustained control. PMID- 19286240 TI - Degradation of atrazine by cobalt-mediated activation of peroxymonosulfate: Different cobalt counteranions in homogenous process and cobalt oxide catalysts in photolytic heterogeneous process. AB - The degradation of atrazine (ATZ) by cobalt-mediated activation of peroxymonosulfate (PMS) has been studied in this work. For the homogenous process, different cobalt counteranions: cobalt(II) nitrate Co(NO(3))(2), cobalt(II) sulfate CoSO(4), cobalt(II) chloride CoCl(2), and cobalt(II) acetate Co(CH(3)COO)(2), have been examined. The inhibitory effect was observed in the process initiated by CoCl(2). For the pH test, wide range of pH level (2-10) has been investigated. It was found that the higher rates were obtained in the normal pH levels. At extreme pH levels, the process was impeded by inactivation of PMS at acidic pH and prohibited by precipitation at basic pH. On the other hand, the recycling capability of cobalt oxide and the oxidative potential of cobalt immobilized titanium dioxide Co-TiO(2) catalyst were analyzed in the heterogeneous process. It was found that the higher the cobalt content in the catalyst, the better the removal performance was resulted. At last, the Co-TiO(2) catalyst synthesized in this work was found to be very effective in transforming ATZ as well as its intermediate in the presence of UV-vis irradiation. PMID- 19286241 TI - Protection of palak (Beta vulgaris L. var Allgreen) plants from ozone injury by ethylenediurea (EDU): roles of biochemical and physiological variations in alleviating the adverse impacts. AB - Ameliorative effects of ethylenediurea (N-[2-(2-oxo-1-imidazolinidyl) ethyl]-N' phenylurea, abbreviated as EDU) against ozone stress were studied on selected growth, biochemical, physiological and yield characteristics of palak (Beta vulgaris L. var Allgreen) plants grown in field at a suburban site of Varanasi, India. Mean eight hourly ozone concentration varied from 52 to 73 ppb which was found to produce adverse impacts on plant functioning and growth characteristics. The palak plants were treated with 300 ppm EDU at 10 days after germination at 10 days interval up to the plant maturity. Lipid peroxidation in EDU treated plants declined significantly as compared to non-EDU treated ones. Significant increment in F(v)/F(m) ratio in EDU treated plants as compared to non-EDU treated ones was recorded. EDU treated plants showed significant increment in ascorbic acid contents and reduction in peroxidase activity as compared to non-EDU treated ones. As a result of the protection provided by EDU against ozone induced stress on biochemical and physiological characteristics of palak, the morphological parameters also responded positively. Significant increments were recorded in shoot length, number of leaves plant(-1), leaf area and root and shoot biomass of EDU treated plants as compared to non-EDU treated ones. Contents of Na, K, Ca, Mg and Fe were higher in EDU treated plants as compared to non-EDU treated ones. The present investigation proves the usefulness of EDU in partially ameliorating ozone injury in ambient conditions. PMID- 19286242 TI - Cr(VI) reduction by an Aspergillus tubingensis strain: role of carboxylic acids and implications for natural attenuation and biotreatment of Cr(VI) contamination. AB - Experiments conducted in shake flask cultures, in minimal medium of pH 5.3 containing 50 microg mL(-1)Cr(VI) with glucose as a carbon source, indicated that the biomass of Aspergillus sp. strain Ed8, a chromate-tolerant fungal strain previously isolated from a chromium-polluted soil, responds to the presence of citrate in the medium by increasing the rate of Cr(VI) reduction; this effect required the use of live biomass and was not observed in medium with lactate. Other natural carboxylic acids or non-natural metal chelating agents showed a stimulatory effect of Cr(VI) reduction by Ed8 biomass; salicylate, tartrate and citrate were the stronger stimulators of the specific rate of Cr(VI) reduction, with about 12, 8 and 7-fold stimulatory effects, respectively, as compared to control cultures without additions. A procedure for Cr(VI) removal from a diluted electroplating effluent was devised, based on the use of growth medium amended with citrate or a mixture of salycilate-tartrate and cycles of recharge of growth medium-diluted effluent. In addition, conditions were adjusted in a 2-L bioreactor to reach a 20-fold increase in the volume of the reduction system with no loss of efficiency. Strain Ed8 was identified as an Aspergillus tubingensis isolate (included in Aspergillus niger species complex) on the basis of the ITS1 5.8s rDNA-ITS2 sequence similarity. PMID- 19286243 TI - Bioavailability and detoxification of cationics: II. Relationship between toxicity and CEC of cationic surfactants on Caenorhabditis elegans (Nematoda) in artificial and natural substrates. AB - The toxicity of the dialkyl quat, didecyldimethylammonium bromide (DDAB), was used as a typical quaternary ammonium compound in studies investigating the role of sorption in reducing DDAB bioavailability in sediment and soil for natural and artificial substrates. Fatty acid derivatives are known to interact ionically with negative charged particles such as clays, humic and fulvic acids, dramatically reducing their bioavailability. Sorption potential was measured using cationic exchange capacities (CEC). The CEC of the substrates was correlated with toxicity of DDAB to the nematode Caenorhabditis elegans considered to be representative of soil and sediment dwelling, free-living nematodes in terms of its sensitivity, size and feeding strategy. Decreased toxicity was found with increasing CEC for both laboratory and field substrates when tested under both soil and sediment conditions and QSARs developed. Testing under soil or sediment conditions had less impact on the toxicity than the CEC of the soil/sediment or whether the substrate was artificial or natural. Habitat preferences were observed during a test in which nematodes were placed into substrates with different CECs. The worms favoured mid-range CECs. Similar preference behaviour may be expected in the environment and a threshold CEC for likely presence of nematodes in a substrate is proposed. Coupled with the substrate toxicity QSAR, threshold CEC preference can be used to provide a no observed effect concentration for DDAB. Expressed as a molar fraction of the CEC, the QSAR obtained for DDAB may be extrapolable to other fatty amine derivatives. If supported by further experimentation and complemented with data from other sediment and soil dwellers the QSAR and threshold CEC value can be validated for use in future regulatory risk assessments of fatty amine derivatives. PMID- 19286244 TI - Profiles and seasonal distribution of airborne fungi in indoor and outdoor environments at a French hospital. AB - A one-year prospective survey of fungal air contamination was conducted in outdoor air and inside two haematological units of a French hospital. Air was sampled with a portable Air System Impactor. During this period of survey, the mean viable fungal load was 122.1 cfu/m(3) in outdoor air samples, and 4.1 and 3.9 cfu/m(3) in samples from adult and pediatric haematology units, respectively. In outdoor samples, Cladosporium was the dominant genus (55%) while in the clinical units, Penicillium sp. (23 to 25%), Aspergillus sp. (15 to 23%) and Bjerkandera adusta (11 to 13%) were the most frequently recovered airborne fungi. The outdoor fungal load was far higher in autumn (168 cfu/m(3)), spring (110 cfu/m(3)) and summer (138 cfu/m(3)) than in winter (49 cfu/m(3)). In indoor air, fungal concentrations were significantly lower in winter (2.7 to 3.1 cfu/m(3)) than in summer (4.2 to 5.0 cfu/m(3)) in both haematology units. In the outdoor environment, Penicillium sp. and Aspergillus sp. were more abundant in winter while the levels of Cladosporium were lowest during this season. In the haematological units, the presence of Aspergillus sp. was stable during the year (close to 20%), Bjerkandera sp. was particularly abundant in winter (close to 30%); levels of Penicillium sp. were highest in autumn while levels of Cladosporium sp. were highest in spring and summer. PMID- 19286245 TI - Are total prostate-specific antigen serum levels in cirrhotic men different from those in normal men? AB - OBJECTIVES: To determine the serum total prostate-specific antigen (tPSA) levels in cirrhotic men and compare them with those in noncirrhotic men. METHODS: We prospectively evaluated 113 cirrhotic patients listed for liver transplantation using the serum tPSA, total testosterone level, and Child-Pugh liver function score according to age and severity of liver disease. The tPSA levels were compared with those of 661 healthy men. The Mann-Whitney U test was used for statistical analysis, with a significance level of .05. RESULTS: The median age of the cirrhotic and noncirrhotic patients was 55 years (range 28-70) and 58 years (range 46-70), respectively (P < .01). However, when stratified by age group (<49, 50-59, and >60 years), this difference was not significant. The median serum tPSA level was 0.3 ng/mL (range 0.04-9.9) and 1.3 ng/mL (range 0.04 65.8) in the cirrhotic and noncirrhotic group, respectively (P < .0001). Stratifying both groups according to age, the cirrhotic patients had significantly lower tPSA levels than did the noncirrhotic patients. According to the Child-Pugh score (A, B, and C), Child-Pugh class C patients had significantly lower tPSA levels than did Child-Pugh class A patients and also had lower testosterone levels than did Child-Pugh class A and B patients. The tPSA levels correlated significantly with the testosterone levels in the cirrhotic patients (P = .028). CONCLUSIONS: The results of our study have shown that cirrhotic patients have approximately 4 times lower serum tPSA levels than noncirrhotic men. Patients with more severe liver disease have lower tPSA and testosterone levels than patients less affected. The tPSA levels in cirrhotic men are affected by the total testosterone levels. PMID- 19286246 TI - Prevalence of urovagina and its effects on reproductive performance in Holstein cows. AB - Two hundred fifty-eight Holstein cows (n=344 lactations) from seven dairy herds were examined every month by vaginoscopy and transrectal palpation to determine the prevalence of urovagina, to investigate its effects on fertility, and to determine some risk factors for urovagina. Accumulation of urine in the vagina of a cow was diagnosed as urovagina and was classified into mild (urine only on the floor of vagina), moderate (urine covering less than or equal to half portion of the external cervical os), or severe (urine covering more than half or whole portion of the external cervical os). In total, 26.7% of lactations had urovagina (mild 11.3%, moderate 11.0%, and severe 4.4%). Survival analysis was used to derive the case definition of clinically relevant urovagina based on the factors associated with increased time to conception. Only moderate (hazard ratio [HR]=0.39) and severe (HR=0.20) degrees of urovagina were associated with reduction in pregnancy rate and thus defined as clinically relevant urovagina. Cows with clinically relevant urovagina had decreased AI submission rate (HR=0.52) and pregnancy rate (HR=0.35), required more inseminations per conception (5 vs. 2; P<0.001), had more days open (370 vs. 136; P<0.001), and were more likely not to get pregnant by 210 d postpartum (odds ratio [OR]=6.62) and to be culled for any reasons (OR=5.32) or for reproductive reasons (OR=9.54) compared with those in cows with no urovagina. Cows with clinically relevant urovagina had a higher risk of endometritis (36.4% vs. 9.2%; P<0.001) compared with that in cows without urovagina. Cow, low BCS at first postpartum examination (OR=2.85), endometritis within 60 d (OR=2.50), and horizontal vulva (OR=9.30) were risk factors for urovagina. In conclusion, 15.4% of lactations had clinically relevant urovagina that increased the risk of endometritis and had detrimental effects on fertility. Individual susceptibility, low BCS, and horizontal vulva increased the risk of diagnosing urovagina in Holstein cows. PMID- 19286247 TI - Macro-, micro- and nano-mechanical investigations on silorane and methacrylate based composites. AB - OBJECTIVES: Cationic ring-opening monomers with a reduced shrinkage stress- silorane--are being used as matrix resins for recently introduced dental composites. Since shrinkage and mechanical properties are generally opposite properties, our study aimed to analyze the mechanical behavior of the silorane based composite in comparison to six homologous clinical successful methacrylate based composites. METHODS: Mechanical properties were determined at macro- (strength and modulus of elasticity), micro- (hardness, modulus of elasticity and creep) and nano-scale (nano-dynamic-mechanical parameters: complex, storage, loss modulus and tandelta) after storing the materials for 24 h in distilled water as well as after aging (thermocycling and storing for 4 weeks in water, saliva or alcohol). RESULTS: The strongest influence on the mechanical properties at macroscopic scale was exerted by the storage media although the influence of the material was also significant. At micro- and nano-scale, the material itself influenced the mechanical properties stronger than the storage. By comparing the four storage conditions, a multivariate analysis revealed a significant decrease in all measured mechanical properties by storing for 4 weeks in water, saliva or alcohol, when compared to the 24 h storage. The strongest effect was exerted through the alcohol storage, whereas the effect of storing for 4 weeks in water or saliva was comparable. The nano-composites were more degraded by a 4-week storage in water and saliva than the micro-hydrids, whereas the effect of storing in alcohol was material depended. The tested silorane-based composite revealed good mechanical properties. The material was very stable in all applied solvents and more reliable by storing in alcohol solutions than the measured methacrylate based composites. SIGNIFICANCE: From the point of view of the mechanical properties measured at macro-, micro- and nano-scale, the silorane-based composite was comparable to clinically successful methacrylate-based composite materials, encouraging the clinical use of the new composite material. PMID- 19286248 TI - The relationship between Shore hardness of elastomeric dental materials and Young's modulus. AB - OBJECTIVES: Hardness of elastomers can be directly related to Young's modulus, a relationship that was investigated in detail by Gent in a paper in 1958. The aim of this study was to test this relationship for 13 dental elastomers (12 silicone and 1 polyether) using the equation derived by Gent and one from BS 903 (1950) that accounts for departures at low values. METHODS: The dental elastomers were subjected to tensile testing and Shore A scale hardness measurements. Young's moduli were calculated from the hardness values using the Gent equation and the BS 903 equation. These calculated values were then compared with values derived experimentally from the tensile tests. RESULTS: Hardness values were in the range 30.2 (+/-0.5)-62.9 (+/-0.8) with the corresponding calculated modulus values in the range 1.1-4.1MPa and 0.9-4.3MPa for the Gent and modified equations, respectively. Young's modulus values derived from the tensile data were in the range 0.8 (+/-0.3)-4.1 (+/-0.3)MPa, showing good agreement with those calculated from the hardness values. Providing viscoelastic creep is minimal during the duration of the test, there is a reasonably well-defined relationship between Shore hardness and Young's modulus in the hardness range studied. SIGNIFICANCE: Simple, non-destructive hardness measurements can be used to determine Young's modulus values. Such values are needed in any calculations of stress distributions in soft lining materials, e.g. by FEA. PMID- 19286249 TI - Using the Identity Processing Style Q-Sort to examine identity styles of Turkish young adults. AB - This paper reports on two studies with Turkish young adults that used the Turkish version of the Identity Processing Style Q-Sort (IPSQ). The IPSQ is based on Berzonsky's informational, normative, and diffused identity styles. Participants sort descriptors of the styles into columns ranging from most to least like them. Patterns in Turkish young adults' identity styles were examined in the first study. The construct validity of the IPSQ-Turkish version based on associations between the IPSQ and measures of cultural orientation, self-definitions, self concept clarity, self-esteem, and coping was tested in the second study. Results of the first study were consistent with the current literature in terms of a negative correlation between informational and diffuse/avoidant styles, and a relatively weaker relationship between informational and normative styles. Support for the construct validity of the Turkish version of the IPSQ was found in the second study, with results consistent with other IPSQ research. PMID- 19286250 TI - In vitro engineering of fibrocartilage using CDMP1 induced dermal fibroblasts and polyglycolide. AB - This study was designed to explore the feasibility of using cartilage-derived morphogenetic protein-1 (CDMP1) induced dermal fibroblasts (DFs) as seed cells and polyglycolide (PGA) as scaffold for fibrocartilage engineering. DFs isolated from canine were expanded and seeded on PGA scaffold to fabricate cell/scaffold constructs which were cultured with or without CDMP1. Proliferation and differentiation of DFs in different constructs were determined by DNA assay and glycosaminoglycan (GAG) production. Histological and immunohistochemical staining of the constructs after being in vitro cultured for 4 and 6 weeks were carried out to observe the fibrocartilage formation condition. The fibrocartilage specific gene expression by cells in the constructs was analyzed by real-time PCR. It was shown that in the presence of CDMP1 the proliferation and GAG synthesis of DFs were significantly enhanced compared to those without CDMP1. Fibrocartilage-like tissue was formed in the CDMP1 induced construct after being cultured for 4 weeks, and it became more matured at 6 weeks as stronger staining for GAG and higher gene expression of collagen type II was observed. Since only weak staining for GAG and collagen type II was observed for the construct engineered without CDMP1, the induction effect on the fibrocartilage engineering can be ascertained when using DFs as seed cells. Furthermore, the potential of using DFs as seed cells to engineer fibrocartilage is substantiated and further study on using the engineered tissue to repair fibrocartilage defects is currently ongoing in our group. PMID- 19286251 TI - X-ray microcomputed tomography for the measurement of cell adhesionand proliferation in polymer scaffolds. AB - We have explored the use of X-ray microcomputed tomography (microCT) for assessing cell adhesion and proliferation in polymer scaffolds. Common methods for examining cells in scaffolds include fluorescence microscopy and soluble assays for cell components such as enzymes, protein or DNA. Fluorescence microscopy is generally qualitative and cannot visualize the scaffold interior. Soluble assays quantitatively measure cell number but do not yield information on cell spatial distribution. Herein, the ability of microCT to detect cells in scaffolds was compared with fluorescence microscopy and a soluble DNA assay. Comparisons were performed using polymer scaffolds that were seeded with cells at different densities and cultured for different times. The results showed that fluorescence microscopy had better resolution than muicroCT and that the soluble DNA assay was approximately 5x more sensitive than microCT under the conditions tested. However, microCT was able to image through opaque scaffolds to yield quantitative 3D imaging and analysis via a single, non-invasive modality. Quantitative microCT analysis of cell penetration into scaffolds was demonstrated. Further, quantitative microCT volume analysis required that the cell density in the scaffolds be greater than 1 million cells per mL indicating that microCT is best suited for quantifying cells at relatively high density during culture in scaffolds. In sum, the results demonstrate the benefits and limitations of using microCT for 3D imaging and analysis of cell adhesion and proliferation in polymer scaffolds. PMID- 19286252 TI - Biocompatibility of biodegradable semiconducting melanin films for nerve tissue engineering. AB - The advancement of tissue engineering is contingent upon the development and implementation of advanced biomaterials. Conductive polymers have demonstrated potential for use as a medium for electrical stimulation, which has shown to be beneficial in many regenerative medicine strategies including neural and cardiac tissue engineering. Melanins are naturally occurring pigments that have previously been shown to exhibit unique electrical properties. This study evaluates the potential use of melanin films as a semiconducting material for tissue engineering applications. Melanin thin films were produced by solution processing and the physical properties were characterized. Films were molecularly smooth with a roughness (R(ms)) of 0.341 nm and a conductivity of 7.00+/-1.10 x 10(-5)S cm(-1) in the hydrated state. In vitro biocompatibility was evaluated by Schwann cell attachment and growth as well as neurite extension in PC12 cells. In vivo histology was evaluated by examining the biomaterial-tissue response of melanin implants placed in close proximity to peripheral nerve tissue. Melanin thin films enhanced Schwann cell growth and neurite extension compared to collagen films in vitro. Melanin films induced an inflammation response that was comparable to silicone implants in vivo. Furthermore, melanin implants were significantly resorbed after 8 weeks. These results suggest that solution processed melanin thin films have the potential for use as a biodegradable semiconducting biomaterial for use in tissue engineering applications. PMID- 19286253 TI - New insights into the role of the tuberous sclerosis genes in leukemia. AB - The genes TSC1 and TSC2, encoding hamartin and tuberin, respectively, have been shown to be involved in the development of the autosomal dominantly inherited tumor syndrome tuberous sclerosis (TSC). However, inactivation of these genes has also been demonstrated to be associated with sporadic bladder cancer, ovarian and gall bladder carcinoma, non-small-cell carcinoma of the lung, breast cancer, pancreatic cancer, astrocytoma, xanthoastrocytoma, ependymomas, oral squamous cell carcinoma and endometrial cancer. The hamartin/tuberin protein complex plays a central role in the regulation of the mammalian target of rapamycin (mTOR) signalling network. A wide variety of components of the mTOR cascade have been demonstrated to be involved in many different human cancers. Mutations in several mTOR pathway component genes are known to cause specific monogenic human genetic diseases and this signalling cascade has been shown to be of relevance for Alzheimer's disease, type 2 diabetes, obesity and hypertrophy. Consequently, e.g. clinical trials for the treatment with rapamycin, a negative regulator of mTOR, of hamartomas in TSC have already been initiated. Now the first evidence is provided for an involvement of the TSC genes in acute leukemia. PMID- 19286254 TI - Prognostic [corrected] significance of angiogenic/lymphangiogenic, anti apoptotic, inflammatory and viral factors in 88 cases with diffuse large B cell lymphoma and review of the literature. AB - BACKGROUND: Diffuse large B cell lymphoma (DLBCL) is the most common subtype of Non-Hodgkins lymphomas (NHL). The outcome of these patients shows a wide variation. We evaluated the effect of six biologic parameters including Cyclooxygenase-2 (Cox-2), Survivin, Epstein Barr Virus (EBV), Vascular Endothelial Growth Factor-A (VEGF-A), Vascular Endothelial Growth Factor-C (VEGF C), Thrombospondin-1 (TSP-1) and clinical parameters. PATIENTS AND METHODS: A follow up study was conducted and 88 cases with DLBCL were included in the study. The data of 72 patients were eligible for the survival analyses. Immunohistochemistry was used to detect these parameters. RESULTS: The ratio of positive cases for Cox-2, VEGF-A, Survivin, VEGF-C, EBV, TSP-1 were 71.6%, 64.8%, 60.2%, 36.4%, 21.6%, 14.8%, respectively. Survivin (+) cases showed higher LDH levels and VEGF-A (+) cases showed higher beta 2 microglobulin (B2M) levels compared with (-) cases. Mean survival rates were found to be significantly shorter in cases expressing VEGF-A, VEGF-C, EBV and Survivin than cases not expressing these. EBV expression (HR: 3.78; 95%CI: 1.47-9.74; p=0.006), VEGF-C expression (HR: 3.22; 95%CI: 1.07-9.68; p=0.037) and extranodal involvement (HR: 3.02; 95%CI: 1.01-8.97; p=0.047) were found to be independent risk factors for prognosis according to the Cox regression analysis. COMMENT: Lymphangiogenesis (VEGF-C) and EBV related viral lymphomagenesis have been found to be related with prognosis in DLBCL patients. PMID- 19286255 TI - Infant acute bilineal leukemia. AB - Most cases of acute leukemia can be assigned to the myeloid, B or T lineage. There are rare cases of acute leukemia, which cannot be clearly classified, because either blasts express antigens of more than one lineage (acute biphenotypic leukemias) or distinct blast populations of two lineages co-exist (acute bilineal leukemias, aBLL). We present a 10-month-old infant with de novo aBLL, characterized by blasts of monocytic and B-cell precursor lineages. All leukemic cells harbored identical complex MLL gene rearrangement. Despite poor initial response, both to acute lymphoblastic leukemia (ALL) induction treatment and acute myeloid leukemia induction blocks, the child reached complete clinical remission with minimal residual disease negative status and was transplanted. Unfortunately, 16 months from HSCT the patient experienced BM relapse with all blasts characterized by pro-B-ALL immunophenotype. This case report illustrates that aBLL is a very aggressive type of acute leukemia that should be individually treated and monitored, particularly in children less than 1 year of age. PMID- 19286256 TI - Dasatinib in the management of lymphoid blast crisis of Philadelphia-positive chronic myeloid leukemia with multiple extra-medullary and intracranial localizations. PMID- 19286257 TI - Physiological changes in reproductively active rainbowfish (Melanotaenia fluviatilis) following exposure to naphthalene. AB - Naphthalene makes up a substantial fraction of polycyclic aromatic hydrocarbons (PAHs) in crude oil and is an important by-product of industry; however, few studies have investigated the toxicity of naphthalene to aquatic organisms. We examined the toxicity of increasing concentrations (0, carrier control, 130, 200 and 400microg/l) of naphthalene to adult rainbowfish (Melanotaenia fluviatilis) for 3 and 14 days to determine its potential to act as an endocrine disruptor. After exposure for 3 days, no changes in sex steroids were measured. After 14 days, a decrease of serum estradiol in females and an increase in serum testosterone in males was observed. These results suggest that naphthalene has the potential to act as an endocrine disruptor, although since no changes in plasma vitellogenin concentrations were observed in either sex, it is unlikely that naphthalene is acting as a xenoestrogen. There was a positive correlation between the incidences of deformities in larval offspring with increasing naphthalene concentrations, suggesting parental transfer of the toxicant. Egg production, hatchability and larval lengths remained unaltered, whilst few changes were measured in gamma-glutamyltranspeptidase (GTP), an enzymatic indicator of spermatogenesis. Contrary to other PAHs, hepatic ethoxyresorufin-O deethylase (EROD) activities declined with increasing exposure concentration, suggesting that naphthalene was either having a cytotoxic effect or disrupting enzyme synthesis. PMID- 19286258 TI - Sublethal cyfluthrin toxicity to carp (Cyprinus carpio L.) fingerlings: biochemical, hematological, histopathological alterations. AB - This study measures a whole series of biological endpoints, and histopathological and behavioral changes of carp (Cyprinus carpio L.) fingerlings in order to investigate the adverse effects of a sublethal concentration of 10 microg/L cyfluthrin for 48h and one week using the semi-static test system. The most significantly altered findings of this study was that total antioxidant status levels decreased in 48h, chloride levels decreased in one week, and sodium and phosphorous levels increased in the 48h-exposed group. Plasma cortisol increased in both the 48h-exposed and the one-week-exposed groups. The unique tissue was the brain that had an increased level of malondialdehyde after one week of exposure. Such biomarkers may be used in assessing adverse/toxic effects of pesticides as environmental stressors. Potential oxidative-stress-inducing effect of synthetic pyrethroid cyfluthrin, as lipid peroxidation, in addition to its sodium channel blocking, is reported for the first time as an additional mechanism of action. PMID- 19286259 TI - The role of the activating receptor NKG2D in autoimmunity. AB - T and NK cell subsets play a crucial role in the immune surveillance designed to fend off dangerous pathogens while leaving endogenous cells untouched. However, occasionally T cells do initiate an attack against self and are therefore termed autoreactive. Moreover, both NK and T cells can be stimulated by danger signals released by cells under "stress", i.e. infection, ongoing necrotic cell death, etc. Consequently, the inappropriate triggering of danger signals, or a failure to switch these off once the immune response has been resolved, can have serious consequences for the host. Ligands for the activating receptor NKG2D (natural killer group 2 member D) are such key danger signals that are presented by "stressed" cells. In this review, we discuss the current knowledge on NKG2D and its ligands in the context of autoimmune diseases and immune-mediated diseases that inadvertently target endogenous cells and/or tissue. PMID- 19286260 TI - A region of the N-terminal domain of meningococcal factor H-binding protein that elicits bactericidal antibody across antigenic variant groups. AB - Meningococcal factor H-binding protein (fHbp) is a promising vaccine antigen. Previous studies described three fHbp antigenic variant groups and identified amino acid residues between 100 and 255 as important targets of variant-specific bactericidal antibodies. We investigated residues affecting expression of an epitope recognized by a murine IgG2a anti-fHbp mAb, designated JAR 4, which cross reacted with fHbps in variant group 1 or 2 (95% of strains), and elicited human complement-mediated, cooperative bactericidal activity with other non bactericidal anti-fHbp mAbs with epitopes involving residues between 121 and 216. From filamentous bacteriophage libraries containing random peptides that were recognized by JAR 4, we identified a consensus tripeptide, DHK that matched residues 25-27 in the N-terminal domain of fHbp. Since DHK was present in both JAR 4-reactive and non-reactive fHbps, the tripeptide was necessary but not sufficient for reactivity. Based on site-directed mutagenesis studies, the JAR 4 epitope could either be knocked out of a reactive variant 1 fHbp, or introduced into a non-reactive variant 3 protein. Collectively, the data indicated that the JAR 4 epitope was discontinuous and involved DHK residues beginning at position 25; YGN residues beginning at position 57; and a KDN tripeptide that was present in variant 3 proteins beginning at position 67 that negatively affected expression of the epitope. Thus, the region of fHbp encompassing residues 25-59 in the N-terminal domain is important for eliciting antibodies that can cooperate with other anti-fHbp antibodies for cross-reactive bactericidal activity against strains expressing fHbp from different antigenic variant groups. PMID- 19286261 TI - Complement-mediated tumour growth: implications for cancer nanotechnology and nanomedicines. AB - The recent unexpected observation that complement activation helps tumour growth and progression has an important bearing on the future development of cancer nanomedicines for site-specific tumour targeting as these entities are capable of triggering complement. These issues are discussed and suggestions are provided for future design and development of safer and effective cancer nanomedicines. PMID- 19286262 TI - Mathematical analysis of clinical data reveals a homunculus of bacterial mimotopes protecting from autoimmunity via oral tolerance in human. AB - Oral tolerance (OT) means systemic immunological unresponsiveness to harmless antigens present in the gastrointestinal tract. We presumed that tolerance to these antigens may also protect self-proteins that show immunological similarity to the intestinal normal flora. To investigate the existence and in vivo relevance of such a tolerogenic molecular mimicry, we focused our attention to Autoimmune Polyendocrine Syndrome type 1 (APS1) and Hemolysis, Elevated Liver Enzymes, Low Platelet count (HELLP) syndrome. APS1 is a human form of Autoimmune Regulator (AIRE) dysfunction with severely impaired central immunotolerance to a specific set of autoantigens, allowing investigation of tolerogenic mimicry by itself, without a disturbing background. HELLP syndrome is a mediocre manifestation of thrombotic microangiopathy, complicating pregnancy, with platelet-fibrin deposits in small blood vessels and transient development of autoantibodies. Impaired microcirculation in the liver is well described, while intestinal ischemia is possible but has not yet been studied. As the harmless nature of an antigen is essential for OT, ischemia-induced bacterial microinvasion represses this process. In case that oral tolerance to a bacterial homunculus is an existing way of self-protection and has an in vivo relevance when central tolerance is intact, significant intestinal ischemia--if present--is expected to promote autoimmunity in HELLP syndrome. We used an experimentally validated, highly reliable mathematical algorithm to predict the extent of immunological similarity between a certain autoantigen and intestinal bacteria. We found a strong negative correlation between the similarity of autoantigens to intestinal bacteria and the production of specific autoantibodies in APS1 (R= 0.70, P=0.002), while a positive correlation was observed in patients with predominantly the severe/moderately severe form of HELLP syndrome according to Mississippi classification (R=0.94, P=0.005). Autoantigen length inversely correlated with the production of autoantibodies in APS1 (R=-0.68, P=0.004). As a longer chain with more epitopes associates with an increased possibility of mimicry to any proteome, molecular mimicry in general--regarding at least major tissue-specific autoantigens--seems to be rather protective. Our calculations support the hypothesis that OT to an intestinal "bacterial homunculus" is an in vivo relevant mechanism of self-protection in humans, furthermore, HELLP syndrome presumably associates with significant intestinal ischemia and leak, resulting in transient autoimmunity via loss of OT. PMID- 19286263 TI - Duration of antibiotic prophylaxis for cardiac surgery: prospective observational study. AB - OBJECTIVES: To assess the effect of prolonging antibiotic prophylaxis in cardiac surgery. METHODS: Prospective before-after cohort study. In 2004, cefazolin was given pre- and intraoperatively at 1g doses while in 2007 it was continued after surgery for 24h. All consecutive adult patients undergoing coronary artery bypass graft, valve, and/or aortic operations during the study periods were included. The primary outcomes were deep sternal wound infection (DSWI) and mortality. Univariate and multivariate analyses were conducted to assess risk factors for DSWI. RESULTS: 954 patients between 1/2004 and 12/2004 were compared to 424 patients between 1/2007 and 6/2007. In 2007, there were significantly more patients >60yrs., emergency and combined operations and the mean logistic EuroSCORE was higher compared to 2004 (8.53% vs. 6.92%, p=0.006). The rate of DSWI decreased non-significantly from 3.8% (36/954) in 2004 to 2.6% (11/424) in 2007, p=0.27. The adjusted odds ratio of the study period for DSWI was 0.89 (95% confidence interval 0.70-1.13). There was no difference in 30-day (5.2% vs. 5.4%) or 6-month mortality (9.2% in both periods), despite increasing patients' risk. CONCLUSIONS: Increasing the duration of antibiotic prophylaxis did not result in a significant decrease in DSWI. The value of prolonging antibiotic prophylaxis after cardiac operations should be further evaluated. PMID- 19286264 TI - Subcutaneous abscess caused by Mycobacterium conceptionense in an immunocompetent patient. PMID- 19286265 TI - Transcranial direct current stimulation in severe, drug-resistant major depression. AB - BACKGROUND: Though antidepressant drugs are the treatment of choice for severe major depression, a number of patients do not improve with pharmacologic treatment. This study aimed to assess the effects of transcranial direct current stimulation (tDCS) in patients with severe, drug-resistant depression. METHODS: Fourteen hospitalized patients aged 37-68, with severe major depressive disorder according to DSM-IV.TR criteria, drug resistant, with high risk of suicide and referred for ECT were included. Mood was evaluated using the Beck Depression Inventory (BDI), the Hamilton Depression Rating Scale (HDRS) and the Visual Analogue Scale (VAS). We also administered cognitive tasks to evaluate the possible cognitive effects on memory and attention. tDCS was delivered over the dorsolateral prefrontal cortex (DLPC) (2 mA, 20 min, anode left, cathode right) twice a day. RESULTS: After five days of treatment although cognitive performances remained unchanged, the BDI and HDRS scores improved more than 30% (BDI p=0.001; HDRS p=0.017). The mood improvement persisted and even increased at four (T2) weeks after treatment ended. The feeling of sadness and mood as evaluated by VAS improved after tDCS (Sadness p=0.007; Mood p=0.036). CONCLUSIONS: We conclude that frontal tDCS is a simple, promising technique that can be considered in clinical practice as adjuvant treatment for hospitalized patients with severe, drug-resistant major depression. PMID- 19286266 TI - Dysfunctional response preparation and inhibition during a visual Go/No-go task in children with two subtypes of attention-deficit hyperactivity disorder. AB - While a response inhibition problem is well-established in children with attention-deficit/hyperactivity disorder of the combined subtype (AD/HDcom), the predominantly inattentive subtype (AD/HDin) has not been investigated previously. This study examined control versus subtype differences in visually evoked response inhibition using task performance and event-related potential (ERP) measures. Children with AD/HDcom (n=15) and AD/HDin (n=15) and age-matched controls (n=15) performed a cued visual Go/Nogo task requiring either activation or inhibition (30%) of a button-press response to the S2 (Go or Nogo stimulus) following the S1 (warning stimulus), presented 1380 ms earlier. Task performance and ERP indices of Warning, Go and Nogo stimulus processing, as well as preparation during the S1-S2 interval, were examined for group differences. Behavioural results indicated a response inhibition deficit in children with AD/HDcom and AD/HDin, with additional response activation problems in AD/HDcom. Topographic ERP differences between controls and both clinical groups suggested atypical (a) preparation for S2 as indexed by the late CNV, (b) early sensory/attentional processing of both S1 and S2, and (c) response inhibition as indexed by N2 and P3. In addition to replicating previous AD/HDcom findings, these results indicate that children with AD/HDin differ from controls in response preparation and inhibition during a cued visual Go/Nogo task. PMID- 19286267 TI - The complement factor C5a receptor is upregulated in NFL-/- mouse motor neurons. AB - In NFL-/- mice, a model of motor neuron degeneration in ALS, degenerating spinal motor neurons express high levels of the receptor for the C5a anaphylatoxin (C5aR) early in the disease process. C5a is a potent in vitro neurotoxin for both Neuro2A and NGF-differentiated PC12 cells. While no interaction was observed between glutamate and C5a, both C5a and kainate upregulated the expression of activated C5aR. C5aR expression was increased in motor neurons in ALS. This data suggests that the early upregulation of C5aR may contribute to motor neuron damage that potentiates excitotoxicity in ALS. PMID- 19286268 TI - Avoiding airway obstruction after pediatric adenotonsillectomy. AB - OBJECTIVE: To evaluate the efficacy of a protocol designed to prevent post adenotonsillectomy airway obstruction in small children with obstructive sleep apnea. DESIGN: Computerized retrospective review of single surgeon case series. SETTING: Tertiary children's medical center. METHODS: Children with sleep study proven obstructive sleep apnea or children under the age of 3 years with clinically suspected obstructive sleep apnea were treated according to a protocol that included: (1) rapid bloodless tonsillectomy; (2) repeated release of the tonsillar retractor; (3) avoidance of uvular edema; (4) routine intra-operative intranasal oxymetazoline, and placement of nasal airway; (5) extended recovery room observation. Primary outcome measures were (1) avoidance of unexpected intensive care unit admission; (2) post-extubation pulmonary edema; (3) aspiration pneumonia. RESULTS: During the period March 2004-August 2007, 864 children underwent adenotonsillectomy by a single surgeon-604 for the indication of obstructive sleep apnea or adenotonsillar hypertrophy with obstruction. Two hundred and ten were under the age of 3 years or had sleep study proven obstructive sleep apnea. There were two unexpected admissions to the pediatric intensive care unit for persistent upper airway obstruction-none required intubation. No child developed post-obstructive pulmonary edema. Three children were treated for infiltrates consistent with aspiration pneumonitis. CONCLUSION: Most cases of post-extubation pulmonary edema and pneumonia can be avoided in young children and those with mild-to-moderate obstructive sleep apnea following a protocol that anticipates and avoids precipitating causes of upper airway obstruction. PMID- 19286269 TI - What's in a name? Another unexplained acronym! PMID- 19286270 TI - Successful desensitization by glargine administration in a patient with insulin allergy: a case report. AB - The patient was a 56-year-old man with type 2 diabetes and insulin allergy. He was administered glargine, which did not produce any allergic reactions, except for a small non-pruritic wheal. Thereafter, other insulin preparation could be administered. We consider this the first case of successful insulin desensitization by glargine administration. PMID- 19286271 TI - Glucose Tranporter-4 expression in monocytes: a systematic review. AB - OBJECTIVE: The purpose of this review was to systematically assess the extent of current knowledge of Glucose Tranporter-4 (GLUT-4) expression in monocytes in humans to address its potential use as a non-invasive and reliable model to investigate the relationships between insulin signalling, GLUT-4 expression and insulin action in vivo. METHOD: Electronic database searches were performed with the keywords 'monocyte', 'leukocyte' and 'white blood cells', and the terms 'GLUT', 'glucose transporter' and 'SLC2A4' (solute carrier family 2 member 4). Studies were examined for robustness of design and outcomes by consensus of three reviewers. RESULTS: Six cross-sectional or observational studies met the criteria for review. Insulin-stimulated GLUT-4 expression in monocytes from subjects likely to have impaired insulin sensitivity appeared blunted relative to healthy subjects. CONCLUSION: The available results provide evidence that monocyte GLUT-4 translocation does occur in response to acute insulin exposure, and may be sensitive to the relative state of insulin resistance of the individual. However, due to the limited quantity and robustness of published data, the ultimate utility of monocyte GLUT-4 expression as an index of whole body insulin responsiveness and the clinical relevance of this methodology is unresolved at this time. PMID- 19286272 TI - Neuro-otologic symptoms in patients with type 2 diabetes mellitus. AB - To assess the prevalence of balance symptoms in type-2 diabetes mellitus (DM2), at the first level of health care. Compared to 101 controls, 101 patients showed a higher frequency of all the symptoms investigated through a standardized questionnaire (p<0.01). In DM2 patients, balance symptoms should be intentionally investigated. PMID- 19286273 TI - pH dependence of persulfate activation by EDTA/Fe(III) for degradation of trichloroethylene. AB - The ability of free ferrous ion activated persulfate (S(2)O(8)(2-)) to generate sulfate radicals (SO(4)(-)) for the oxidation of trichloroethylene (TCE) is limited by the scavenging of SO(4)(-) with excess Fe(2+) and a quick conversion of Fe(2+) to Fe(3+). This study investigated the applicability of ethylene diamine-tetra-acetic acid (EDTA) chelated Fe(3+) in activating persulfate for the destruction of TCE in aqueous phase under pH 3, 7 and 10. Fe(3+) and EDTA alone did not appreciably degrade persulfate. The presence of TCE in the EDTA/Fe(3+) activated persulfate system can induce faster persulfate and EDTA degradation due to iron recycling to activate persulfate under a higher pH condition. Increasing the pH leads to increases in pseudo-first-order-rate constants for TCE, S(2)O(8)(2-) and EDTA degradations, and Cl generation. Accordingly, the experiments at pH 10 with different EDTA/Fe(3+) molar ratios indicated that a 1/1 ratio resulted in a remarkably higher degradation rate at the early stage of reaction as compared to results by other ratios. Higher persulfate dosage under the EDTA/Fe(3+) molar ratio of 1/1 resulted in greater TCE degradation rates. However, increases in persulfate concentration may also lead to an increase in the rate of persulfate consumption. PMID- 19286274 TI - Apocynin-induced nitric oxide production confers antioxidant protection in maize leaves. AB - The effect of apocynin on nitric oxide (NO) synthesis and oxidative stress was studied in corn (Zea mays) seedlings. After treatment with 100 microM apocynin, strongly increased amounts of NO were detected in the leaves. This NO production was reduced by more than 70% by N(G)-nitro-l-arginine methyl ester (L-NAME), a NO synthase (NOS) inhibitor, but there was no reduction in NO production when apocynin was applied in combination with diphenylene iodonium (a plant NOX inhibitor). When maize seedlings were UV-B-irradiated, cellular damage occurred and reactive oxygen species (ROS) were found widely distributed in chloroplasts and mesophyll cells. Pre-treatment with apocynin and coinciding NO accumulation prevented this damage. However, the protective effect was averted by L-NAME application. Leaf discs placed in 1M H(2)O(2) for 24h showed a reduction in chlorophyll content that could also be avoided by apocynin treatment. Our results show that apocynin induces the accumulation of NO in leaves of maize seedlings through a NOS-like activity, a mechanism alternative to NOX inhibition, and confers an augmented tolerance to different types of abiotic oxidative stress. Indeed, we propose the use of apocynin as an alternative approach to study NO functionality in plants. PMID- 19286275 TI - Raffinose and stachyose metabolism are not required for efficient soybean seed germination. AB - Raffinose family oligosaccharides (RFOs), which include raffinose and stachyose, are thought to be an important source of energy during seed germination. In contrast to their potential for promoting germination, RFOs represent anti nutritional units for monogastric animals when consumed as a component of feed. The exact role for RFOs during soybean seed development and germination has not been experimentally determined; but it has been hypothesized that RFOs are required for successful germination. Previously, inhibition of RFO breakdown during imbibition and germination was shown to significantly delay germination in pea seeds. The objective of this study was to compare the germination potential for soybean seeds with either wild-type (WT) or low RFO levels and to examine the role of RFO breakdown in germination of soybean seeds. There was no significant difference in germination between normal and low RFO soybean seeds when imbibed/germinated in water. Similar to the situation in pea, soybean seeds of wild-type carbohydrate composition experienced a delay in germination when treated with a chemical inhibitor of alpha-galactosidase activity (1 deoxygalactonojirimycin or DGJ) during imbibition. However, low RFO soybean seed germination was not significantly delayed or reduced when treated with DGJ. In contrast to the situation in pea, the inhibitor-induced germination delay in wild type soybean seeds was not partially overcome by the addition of galactose or sucrose. We conclude that RFOs are not an essential source of energy during soybean seed germination. PMID- 19286276 TI - Stress is critical for LPS-induced activation of microglia and damage in the rat hippocampus. AB - The hippocampus is insensitive to strong inflammatory stimulus under normal conditions and one of the most severely affected areas in Alzheimer's disease. We have analyzed the effect of chronic stress for 9 days in the hippocampus unilaterally injected with LPS. In non-stressed rats, LPS injection failed to activate microglia although a subset of degenerating cells in the CA1 area was evident. This effect was not accompanied by loss of Neu-N positive neurons in the CA1 area. In stressed rats, LPS injection had a dramatic effect in activating microglia along with astrogliosis and BDNF mRNA induction. NeuN immunostaining demonstrated a loss of about 50% of CA1 pyramidal neurons under these conditions. Fluoro jade B histochemistry demonstrated the presence of degenerating cells in most of CA1 area. Mechanistically, combination of chronic stress and LPS resulted in prominent activation of MAPKs including JNK, p38 and ERK clearly different from LPS injection in controls. Further, LPS+stress induced a dramatic decrease in phosphorylated levels of both Akt and CREB, which fully supports a consistent deleterious state in the hippocampal system under these conditions. Treatment with RU486, a potent inhibitor of glucocorticoid receptor activation, significantly protected animals against the deleterious effects observed in LPS stressed animals. PMID- 19286277 TI - [Climate, longevity and ageing]. PMID- 19286278 TI - [Reflections on practices for early cancer diagnosis in Spain]. PMID- 19286279 TI - [User satisfaction with teleophthalmology with nonmydriatic camera for diabetic retinopathy screening]. AB - OBJECTIVE: To determine satisfaction with the retinography service among patients with diabetes. METHODS: We performed a telephone survey of 64 users from July 2006 to March 2007. The mean age was 65.2 years, 57.8% were men, and 54.7% were from urban primary care centers. The variables analyzed were sex, age, primary care team, retinography/tonometry (normal/pathologic), accessibility, punctuality, hygiene, consultation length, explanations, good hands, kindness on a scale rated bad/average/good/very good/perfect, satisfaction with the telephone call informing users of the results of the examination, and overall satisfaction, both rated on a scale from 0 to 10. RESULTS: Accessibility, punctuality, hygiene, consultation length, good hands and kindness received scores of >80%. The mean overall satisfaction score was 8.38% (95% confidence interval [95%CI]: 8.03 8.72), while satisfaction with the telephone call was 7.88% (95%CI: 7.4-8.36). The variables associated (p<0.05) with overall satisfaction of 8 were consultation length, receiving comprehensible explanations, and the telephone call informing patients of the results of the examination. Logistic regression showed (p<0.05) that the variable with the greatest influence on satisfaction was the telephone call. CONCLUSIONS: the retinography service was favorably evaluated. The variable with the greatest influence on high satisfaction was communicating the results by telephone. The service will promote new technologies (SMS, e-mail). PMID- 19286280 TI - [Effects of influenza vaccination on the percentage of CD4+ T lymphocytes in HIV 1/2 positive patients: a cohort study]. AB - OBJECTIVE: To determine the incidence of immunologic change in the percentage of CD4+ T lymphocytes in HIV 1/2 positive patients in the first quarter after influenza vaccination (P-CIR) and to compare the demographic and clinical characteristics associated with this change. METHODS: We studied 105 patients with HIV/AIDS in a retrospective hospital cohort between 2001-2006. P-CIR was considered as a decrease of >3% in the prevaccination CD4+ percentage. Crude and adjusted OR (sex, age, antiretroviral therapy, clinical stability, prevaccination viremia and prevaccination total CD4) were evaluated by logistic regression (95%CI). RESULTS: The incidence of P-CIR was 33.3%. P-CIR was persistent in 31.4% for 7 months after vaccination regardless of high prevaccination viremia. No association was found between demographic and clinical variables and P-CIR [crude OR: 0.90 (0.17-4.8); adjusted OR: 1.09 (0.17-6.8)]. CONCLUSIONS: The results showed that the immunological change after vaccination was not inconsiderable. However, this change was mainly transient. PMID- 19286281 TI - [Policies to reduce health inequalities in Spain]. PMID- 19286282 TI - Synthesis and anticonvulsant activity of some new bishydrazones derived from 3,4 dipropyloxythiophene. AB - A series of new 3,4-dipropyloxy-N(2),N(5)-bis(substituted)thiophene-2,5 dicarbohydrazides (4-30) were synthesized from ethyl thiodiglycolate and diethyloxalate through multistep reactions. Following Dieckmann-Komppa reaction, the required precursor 3,4-dihydroxythiophene-2,5-diester (1) was prepared. This was derivatized with propyl bromide and further converted to corresponding hydrazide (3), which was finally transformed to targeted hydrazones (4-30) by conventional methods. The newly synthesized compounds were characterized using FT IR, (1)H and (13)C NMR, EI-MS and elemental analyses. The anticonvulsant activity of all the title compounds was investigated against maximal electroshock induced seizures (MES) and subcutaneous pentylenetetrazole (scMET) models and their neurotoxicity was also evaluated. Some of the selected compounds were subjected to 6 Hz test in order to evaluate their uncover activities. Compound 3,4 dipropyloxy-N(2),N(5)-bis[1-(2-thienyl)ethylidene]thiophene-2,5-dicarbohydrazide (15) has emerged as a lead in this series with less neurotoxicity. PMID- 19286284 TI - [An abdominal tumor]. PMID- 19286283 TI - Bibenzyl- and stilbene-core compounds with non-polar linker atom substituents as selective ligands for estrogen receptor beta. AB - A series of structurally simple bibenzyl-diol and stilbene-diol core molecules, structural analogs of the well-known hexestrol and diethylstilbestrol non steroidal estrogens, were prepared and evaluated as estrogen receptor (ER) subtype-selective ligands. Analysis of their ERalpha and ERbeta binding showed that certain substitution patterns engendered binding affinities that were >100 fold selective for ERbeta. When further investigated in cell-based gene transcription assays, some molecules showed similarly high relative transcriptional potency selectivity in favor of ERbeta. Interestingly, the most ERbeta-selective molecules were those bearing non-polar substituents on one of the internal carbon atoms. These compounds should be useful probes for determining the physiological roles of ERbeta, and they might lead to the development of more selective and thus safer pharmaceuticals. PMID- 19286285 TI - [Cholesterol crystal emboli presenting as a mononeuropathy]. AB - Cholesterol crystal embolism is observed in polyvascular patients usually following a medical procedure. We report a 93-year-old patient with lower limb mononeuropathy in whom cholesterol crystal embolism was confirmed by neuromuscular biopsy. Previously, an ophthalmoscopy and a livedo biopsy had been performed and the results were normal. Mononeuropathy simplex or multiplex is a clinical expression for cholesterol crystal embolism that may thus be confounded with polyarteritis nodosa. Neuromuscular biopsy can provide accurate diagnosis if other investigations are inconclusive. PMID- 19286286 TI - [Cardiac involvement in Steinert myotonic dystrophy]. AB - INTRODUCTION: Myotonic dystrophy type 1 (DM1) is an autosomal dominant multisystemic disease with frequent cardiac involvement that may cause sudden death. This study was performed to determine the various cardiac manifestations in DM1, their frequency and the relevance of cardiac electrophysiological study in this disease. METHODS: Ten patients with DM1, five men and five women, mean age 44.3+/-7.8 years underwent neurological and cardiac assessments. RESULTS: The most frequent electrocardiographic findings were conduction abnormalities, essentially by intraventricular conduction defects (eight out of ten cases) such as bundle branch or fascicular blocks. Echocardiography showed alterations in systolic left ventricular function in two cases. Invasive electrophysiology testing showed sub-hisien block in three patients, requiring cardiac pacemaker implantation. These three patients had normal duration of PR interval and normal width of QRS complex. CONCLUSIONS: We recommend that all patients with DM1 should undergo cardiac investigation to detect subclinical cardiac involvement. PMID- 19286287 TI - Attitudes towards mental illness among health care students at Swedish universities--a follow-up study after completed clinical placement. AB - The aim of the study was to examine the changes in attitudes towards mental illness after theoretical education and clinical placement among students from university programmes preparing for different kinds of health professions. Three different questionnaires were used, measuring the level of familiarity with mental illness and attitudes towards mental illness in general and towards specific mental illnesses. The data were collected on two occasions, before the theoretical course and after the completed clinical placement. The result showed that the attitudes toward mental illness in general had changed in a less stigmatising direction after the clinical placement. On the other hand, attitudes toward specific mental illnesses did not show any major changes. A conclusion is that the clinical placement included in the university programmes to some extent could affect attitudes in a de-stigmatizing direction, possibly because of the interaction with persons suffering from mental illness and experienced supervisors. PMID- 19286288 TI - An exploration of the assessment experiences of new academics as they engage with a community of practice in higher education. AB - This paper considers a phenomenological research study that attempted to explore how new academics were introduced to the assessment process within a Higher Education context. Two key educational perspectives have shaped the interpretation of the studies findings. These are Nonaka and Takeuchi's [Nonaka, I., Takeuchi, H., 1995. The Knowledge Creating Company: How Japanese Companies Create the Dynamics of Innovation. Oxford University Press, New York] model of knowledge conversion and Lave and Wenger's work on communities of practice (1991, 2002). Three key findings emerged from this work. Firstly, the study highlights a number of issues relating to the types of support and guidance that new academics receive. These were divided into formal and informal types that either promoted conformity or facilitated challenge. Secondly, the study suggests that the ways in which experienced academic staff communicate their assessment knowledge and interact with new academics may require further consideration. Finally, the study raises questions about the type of academic that the organisation would wish to develop. PMID- 19286289 TI - Endoscopic placement of postpyloric nasoenteric feeding tubes: the importance of the guidewire used. PMID- 19286290 TI - Survey of maxillofacial 'out of hours' operating in the United Kingdom. AB - This survey looks at 'out of hours' maxillofacial operating in the United Kingdom during September 2006 and was sent out to all the maxillofacial consultants in the United Kingdom. Most respondents consider out of hours as '2200-0759'; however many hospitals now routinely work extended lists up to 2200 h. Since nearly 65% of respondents had no dedicated trauma list, the provision of theatre flexibility with prioritized trauma slots which would be used for other purposes depending upon the immediate departmental needs appears to be a viable compromise for the foreseeable future. PMID- 19286291 TI - Contaminants reduce the richness and evenness of marine communities: a review and meta-analysis. AB - Biodiversity of marine ecosystems is integral to their stability and function and is threatened by anthropogenic processes. We conducted a literature review and meta-analysis of 216 studies to understand the effects of common contaminants upon diversity in various marine communities. The most common diversity measures were species richness, the Shannon-Wiener index (H') and Pielou evenness (J). Largest effect sizes were observed for species richness, which tended to be the most sensitive index. Pollution was associated with marine communities containing fewer species or taxa than their pristine counterparts. Marine habitats did not vary in their susceptibility to contamination, rather a approximately 40% reduction in richness occurred across all habitats. No class of contaminant was associated with significantly greater impacts on diversity than any other. Survey studies identified larger effects than laboratory or field experiments. Anthropogenic contamination is strongly associated with reductions in the species richness and evenness of marine habitats. PMID- 19286292 TI - National baseline survey of soil quality in The Netherlands. AB - To determine background values of the 252 chemical compounds listed in Dutch legislation, a survey was designed with the aim of estimating percentiles of the cumulative frequency distributions and areal fractions exceeding the former, legislative reference values. Sampling locations were selected by probability sampling, so that in estimating the target quantities no model assumptions on the spatial variation were required, and valid estimates could be obtained by design based inference. Strata in random sampling were formed by overlaying an aggregated soil map and land use map. For most of the heavy metals the areal fraction with concentrations in the topsoil (0-10 cm) exceeding the reference value was smaller than the allowable maximum of 5%. For these compounds a background value was determined smaller than the reference value. Exceptions were V, Co, Ba and Cu, for which a background value was defined (slightly) larger than the reference value. PMID- 19286293 TI - Do conditions in early life affect old-age mortality directly and indirectly? Evidence from 19th-century rural Sweden. AB - Previous research has shown that the disease load experienced during the birth year, measured as the infant mortality rate, had a significant influence on old age mortality in nineteenth-century rural Sweden. We know that children born in years with very high rates of infant mortality, due to outbreaks of smallpox or whooping cough, and who still survived to adulthood and married, faced a life length several years shorter than others. We do not know, however, whether this is a direct effect, caused by permanent physical damage leading to fatal outcomes later in life, or an indirect effect, via its influence on accumulation of wealth and obtained socio-economic status. The Scanian Demographic Database, with information on five rural parishes in southern Sweden between 1813 and 1894, contains the data needed to distinguish between the two mechanisms. First, the effects of conditions in childhood on obtained socio-economic status as an adult are analyzed, then the effects of both early-life conditions and socio-economic status at various stages of life on old-age mortality. By including random effects, we take into account possible dependencies in the data due to kinship and marriage. We find that a high disease load during the first year of life had a strong negative impact on a person's ability to acquire wealth, never before shown for a historical setting. This means that it is indeed possible that the effects of disease load in the first year of life indirectly affect mortality in old age through obtained socio-economic status. We find, however, no effects of obtained socio-economic status on old-age mortality. While the result is interesting per se, constituting a debatable issue, it means that the argument that early-life conditions indirectly affect old-age mortality is not supported. Instead, we find support for the conclusion that the effect of the disease load in early-life is direct or, in other words, that physiological damage from severe infections at the start of life leads to higher mortality at older ages. Taking random effects at family level into account did not alter this conclusion. PMID- 19286294 TI - Harms and benefits: collecting ethnicity data in a clinical context. AB - Although ethnicity data are collected in most countries at the population level, it has become more common to collect such data in healthcare settings, partially in response to growing health and social inequities worldwide. However, the implications of doing so have not been studied. This two-year study was designed to critically examine the implications of collecting ethnicity data in healthcare settings. Using a critical ethnographic approach, we interviewed 104 patients, community and healthcare leaders, and healthcare workers within diverse clinical contexts in a large city in Western Canada in 2006-2007. This paper presents an interpretive thematic analysis, using an ethical lens, of the harms and benefits associated with the process of data collection in a clinical context. While most leaders and healthcare workers and some patients envisioned potential benefits associated with having ethnicity data, these benefits were seen as largely contingent upon action being taken to ameliorate inequities. Overwhelmingly, however, leaders from ethno-cultural communities and patients of diverse identities anticipated potential harm arising both from having ethnicity data and the process of collection. The analysis illustrates that in today's sociopolitical context, collecting ethnicity data in clinical contexts may engender considerable harm, particularly for racialized, vulnerable patients. If ethnicity data are currently collected at the population level, evidence of benefit is required before proceeding to collect these data at the point of care. PMID- 19286295 TI - Probing the human brain with stimulating electrodes: the story of Roberts Bartholow's (1874) experiment on Mary Rafferty. AB - Roberts Bartholow's 1874 experiment on Mary Rafferty is widely cited as the first demonstration, by direct application of stimulating electrodes, of the motor excitability of the human cerebral cortex. The many accounts of the experiment, however, leave certain questions and details unexamined or unresolved, especially about Bartholow's goals, the nature and quality of the evidence, and the experiment's role in the history of theory and research on localisation of function. In this article, we try to fill these gaps and to tell the full story. We describe Bartholow's career up to 1874, review the theoretical and empirical background for the experiment, and present Bartholow's own account of the experiment as well as those of his supporters and critics. We then present our own analysis, assess the experiment's influence on contemporaneous scientific opinion about cortical excitability, and trace its citation record into our own time. We also review and assess ethical criticisms of Bartholow and their effects on his career, and we close by discussing the role we think the experiment deserves to play in the history of theory and research on cortical excitability. PMID- 19286296 TI - [Diagnostic yield of culture for mycobacteria in tuberculous pleural effusions]. PMID- 19286297 TI - [Activity of an intermediate respiratory care unit attached to a respiratory medicine department]. AB - BACKGROUND AND OBJECTIVE: With the development of noninvasive ventilation (NIV), patients with increasingly complex needs have been admitted to respiratory medicine departments. For this reason, such departments in Spain and throughout Europe have been adding specialized respiratory intermediate care units (RICUs) for monitoring and treating patients with severe respiratory diseases. The aim of the present study was to describe the activity of such a RICU. The description may be of use in facilitating the setting up of RICUs in other hospitals of the Spanish National Health Service. METHODS: A systematic record of activity carried out in the RICU of the Hospital Universitario Son Dureta between January and December 2006 was kept prospectively. RESULTS: Of 206 patients with a mean (SD) age of 65 (14) years admitted to the unit, 67% came from the emergency department, 14% from the respiratory medicine department, and 12% from the intensive care unit (ICU). The most common admission diagnoses were exacerbated chronic obstructive pulmonary disease (COPD) (n=97, 47.1%), pneumonia (n=39, 18.9%), heart failure (n=17, 8.2%), and pulmonary vascular diseases (n=18, 8.7%). One hundred twenty-one patients (59%) required NIV. Mean length of stay in the RICU was 5 (5) days. Patients were discharged to the conventional respiratory ward in 79.1% of the cases; 7.8% required subsequent admission to the ICU, and 9.7% died. Of the patients with exacerbated COPD (mean age, 66.5 [10] years; mean length of stay, 4.6 [4.5] days), 67% required NIV, 7.2% required subsequent admission to the ICU, and 8.2% died. CONCLUSIONS: The creation of a RICU by a respiratory medicine department is viable in Spain. Such units make it possible to treat a large number of patients with a low rate of therapeutic failures. Exacerbated COPD was the most common diagnosis on admission to our RICU, and the need for NIV the most common criterion for admission. PMID- 19286298 TI - [Effective anti-immunoglobulin-E antibody treatment of a patient with allergic bronchopulmonary aspergillosis]. PMID- 19286299 TI - Do we know the strength of the chorioamnion? A critical review and analysis. AB - The chorioamnion is the membrane that surrounds the fetus during gestation. Normally, it must remain intact for the duration of pregnancy, 37-42 weeks, and only rupture during or just before labour and delivery of the fetus. In a significant number (3%) of all births, this does not happen, and membranes rupture before term, resulting in preterm birth and significant perinatal morbidity. It is known that the material properties of chorioamnion may play a major role in mechanical rupture; a number of studies have been undertaken to characterise the physical nature of chorioamnion and examine factors that may predispose to rupture. However, the existing literature is inconsistent in its choice of both physical testing methods and data analysis techniques, motivating the current review. Experimental data from a large number of chorioamnion mechanical studies were collated, and data were converted to standard engineering quantities. The failure strength of the chorioamnion membrane was found consistently to value approximately 0.9 MPa. It is hoped that past and future studies of membrane mechanics can provide insight into the role of chorioamnion in labour and delivery. In addition, biomechanical approaches can help elucidate the potential causes of early rupture, and suggest future protocols or treatments that could both diagnose and prevent its occurrence. PMID- 19286300 TI - Enhanced prion protein stability coupled to DNA recognition and milieu acidification. AB - The prion protein (PrP) is the major agent involved in the transmissible spongiform encephalopathies (TSEs). Nucleic acids have been reported to bind PrP with high affinity, although the physiopathological roles for recognition are still not clear. In this work we investigate the stability of a soluble, 1:1 complex formed between an 18 base-pair DNA fragment and the full-length murine recombinant prion protein (mrPrP). DNA confers a gain in mrPrP stability against urea and guanidinium denaturation, which is enhanced at lower pHs and in moderate concentrations of NaCl. We discuss the cooperative folding transition coupled to DNA binding and acidification in terms of the possible cellular scenarios found during complex internalization and degradation. PMID- 19286301 TI - Designing of an environmental assessment algorithm for surface mining projects. AB - This paper depicts the method used to quantify the environmental impact of mining activities in surface mine projects. The affected environment was broken down into thirteen components, such as Human health and immunity, Surface water, Air quality, etc. The effect of twenty impacting factors from the mining and milling activities was then calculated for each Environmental Component. Environmental assessments are often performed by using matrix methods in which one dimension of the matrix is the "Impacting Factor" and the other one is the "Environmental Components". For the presented matrix method, each Impacting Factor was first given a magnitude between -10 and 10. These factors are used to set up a matrix named Impacting Factor Matrix, whose elements represent the Impacting Factor values. The effects of each Impacting Factor on each Environmental Component were then quantified by multiplying the Impacting Factor Matrix by Weighting Factor Matrix. The elements of the weighting factors matrix reflect the effects of each Impacting Factor on each Environmental Component. The outlined method was originally developed for a mining and milling operation in Iran, but it can successfully be used for mining ventures and more general industrial activities in other countries in accordance to their environmental regulations and laws. PMID- 19286302 TI - Insignificant prostate cancer and active surveillance: from definition to clinical implications. AB - CONTEXT: Due to early detection strategies, prostate cancer is diagnosed early in its natural history. It remains unclear whether all patients diagnosed with prostate cancer warrant radical treatment or may benefit from delayed intervention following active surveillance. OBJECTIVE: A systematic review of active surveillance protocols to investigate the inclusion criteria for active surveillance and the outcome of treatment. EVIDENCE ACQUISITION: Medline was searched using the following terms: prostate cancer, active surveillance and expectant management for dates up to October 2008. Further studies were chosen on the basis of manual searches of reference lists and review papers. EVIDENCE SYNTHESIS: Numerous studies on active surveillance were identified. The recent inclusion criteria of the studies are rather similar. Keeping the short follow-up of all studies in mind, the majority of men stay on active surveillance, and the percentage of patients receiving active treatment is as high as 35% of all patients. Once a patients requires active treatment, most patients still present with curable prostate cancer. Furthermore, only few deaths due to prostate cancer have occurred. CONCLUSIONS: Active surveillance is an alternative option to immediate treatment of men with presumed insignificant prostate cancer. It seems that criteria used to identify men with low-risk prostate cancer are rather similar, and immediate treatment of men meeting these criteria may result in an unnecessary number of treatments in these highly selected patients. Data from randomised trials comparing active surveillance and active treatment will provide additional insight into outcome and follow-up strategies. PMID- 19286303 TI - Editorial comment on: Insignificant prostate cancer and active surveillance: from definition to clinical implications. PMID- 19286304 TI - Editorial comment on: Insignificant prostate cancer and active surveillance: from definition to clinical implications. PMID- 19286305 TI - Re: Mesut Remzi, Michael Marberger. Renal tumor biopsies for evaluation of small renal tumors: why, in whom, and how? Eur urol 2009;55:359-67. PMID- 19286306 TI - Aberration of the enzymatic activity of Fhit tumor suppressor protein enhances cancer cell death upon photodynamic therapy similarly to that driven by wild-type Fhit. AB - The tumor suppressor Fhit protein lost in many human pre-malignant tissues, possesses diadenosine triphosphate activity regulated by a photosensitizer, protoporphyrin IX (PpIX) in vitro. Interestingly, when exogenously restored, the protein suppresses the growth of human cervical carcinoma HeLa cells which is further enhanced by PpIX. Additionally, Fhit production enhances the overall response of cells to PpIX-mediated photodynamic reaction. In the present study, we have estimated, for the first time, the biological activity of two Fhit mutated forms exhibiting aberrant Ap(3)A hydrolase activity in vitro which emphasizes the recent findings that hydrolysis of Ap(3)A is not necessary for Fhit tumor suppression function. Using several biophysical methods we revealed the dynamic nature of mutant Fhit-PpIX complexes in vitro which support our previous hypothesis that Fhit-Ap(3)A-PpIX might be a signaling molecule driving apoptosis in cancer cells. Moreover, according to our findings, substitution at histidine94 in Fhit active site induces the vulnerability of HeLa cells to PpIX PDT in a similar manner to that caused by wild-type Fhit protein. These results support the view that inhibition of Fhit hydrolase activity might be a crucial element in a Fhit-driven cancer cells death. PMID- 19286307 TI - Indomethacin-loaded nanocapsules treatment reduces in vivo glioblastoma growth in a rat glioma model. AB - Multimodal combinations of target agents with radiation and chemotherapy may enhance cancer treatment efficacy; however, despite these treatments, gliomas recur early due to their highly proliferative, infiltrative and invasive behaviors. Nanoparticles of biodegradable polymers for anticancer drug delivery have attracted intensive interest in recent years since they may provide a sustained, controlled and targeted delivery. In the present study, we investigated the effect of indomethacin-loaded nanocapsules in an experimental glioma model. The rats treated with indomethacin-loaded nanocapsules demonstrated a significant reduction in tumor size and half of these animals presented just cells with characteristics of a residual tumor, as shown by immunostaining for nestin. Pathological analyses showed that the treated gliomas presented a significant reduction in the mitotic index and other histological characteristics that indicate a less invasive/proliferative tumor. An important finding of the present study is that indomethacin carried by polymeric nanocapsules achieved higher intracerebral drug concentrations than those of indomethacin in solution. Furthermore, indomethacin achieved a greater concentration in the hemisphere where the glioma was implanted, compared with the contralateral healthy hemisphere. Indomethacin-loaded nanocapsule treatment did not cause characteristics of toxicity and increased the survival of animals. Thus, our results show that polymeric nanocapsules are able to increase the intratumoral bioavailability of indomethacin and reduce the growth of implanted gliomas. Data suggest that indomethacin-loaded nanocapsules could offer new and potentially highly effective strategies for the treatment of malignant gliomas. PMID- 19286308 TI - Bruceine D induces apoptosis in pancreatic adenocarcinoma cell line PANC-1 through the activation of p38-mitogen activated protein kinase. AB - Pancreatic cancer is a malignant disease with extremely high mortality. Our previous work found that Brucea javanica fruit possesses potent anti-pancreatic cancer activity. In the present study, brucein D (BD), a quassinoid found abundantly in B. javanica fruit, was evaluated for its anti-proliferative and apoptogenic actions. BD inhibited the growth of three pancreatic cancer cell lines, i.e., PANC-1, SW1990 and CAPAN-1, but exerted only modest cytotoxicity on non-tumorigenic Hs68 cells. Hoechst 33342 staining and Cell Death Detection ELISA(PLUS) assay revealed that BD-induced DNA fragmentation in PANC-1 cells. Moreover, subG1 phase was observed in the BD-treated cells. Western blot experiments indicated that BD exposure augmented caspase 3, 8, 9 and bak protein levels, while attenuating the expression of bcl-2. Furthermore, BD treatment promoted phosphorylation of p38-MAPK. The selective p38-MAPK inhibitor SB203580 effectively mitigated the BD-induced apoptosis in PANC-1 cells, suggesting that p38-MAPK signaling pathway was involved in the BD-induced apoptosis in pancreatic cancer cells. Taken together, our results provide experimental evidence to support the traditional use of B. javanica fruit in cancer treatment, and render BD a promising chemical candidate for further development into anti-pancreatic cancer agent. PMID- 19286309 TI - CXCR4 receptor positive spheroid forming cells are responsible for tumor invasion in vitro. AB - Stem cells have been found to be involved in breast cancer growth, but the specific contribution of cancer stem cells in tumor biology, including metastasis, is still uncertain. We found that murine breast cancer cell lines 4T1, 4TO7, 167Farn and 67NR contains cancer stem cells defined by CXCR4 expression and their capability of forming spheroids in suspension culture. Importantly, we showed that CXCR4 expression is essential for tumor invasiveness because both CXCR4 neutralizing antibody and shRNA knockdown of the CXCR4 receptor significantly reduced tumor cell invasion. PMID- 19286310 TI - Action of palmatine on Tetrahymena thermophila BF5 growth investigated by microcalorimetry. AB - Using a thermal activity monitor (TAM) air isothermal microcalorimeter with ampoule mode, the thermo-genic curves of the metabolism of Tetrahymena thermophila BF(5) growth at 28 degrees C were obtained and the action of palmatine on it was investigated. Meanwhile, the biomass change during the process of T. thermophila BF(5) growth coexisted with palmatine was studied by a haemacytometer. The results showed that a low concentration (50 microg/mL) of palmatine began to inhibit the growth of T. thermophila BF(5), and when the concentration of palmatine reached 600 microg/mL, T. thermophila BF(5) could not grow at all. The relationship between the growth rate constant (k) and the concentration c was almost linear with the correlation coefficient of 0.9957, showing the strong toxic action of palmatine on T. thermophila BF(5) growth. The biomass during T. thermophila BF(5) growth decreased obviously by the addition of palmatine at different concentrations. The investigation of biomass agreed well with the results obtained by means of microcalorimetry. PMID- 19286311 TI - Dissolution kinetics of spent petroleum catalyst using sulfur oxidizing acidophilic microorganisms. AB - Bioleaching studies of spent petroleum catalyst were carried out using sulfur oxidizing, Acidithiobacillus species. Leaching studies were carried out in two stage, in the first stage bacteria were grown and culture filtrate was used in the second stage for leaching purpose. XRD analysis of spent petroleum catalyst showed oxides of V, Fe and Al and sulfides of Mo and Ni. The leaching kinetics followed dual rate, initial faster followed by slower rate and equilibrium could be achieved within 7 days. The leaching rate of Ni and V were high compared to Mo. The low Mo leaching rate may be either due to formation of impervious sulfur layer or refractoriness of sulfides or both. The leaching kinetics followed 1st order rate. Using leaching kinetics, rate equations for dissolution process for different metal ions were evaluated. The rate determining step observed to be pore diffusion controlled. PMID- 19286312 TI - Enhanced production of lactic acid with reducing excess sludge by lactate fermentation. AB - The development of a facile technology for utilizing effectively and/or reducing excess sludge is one of the urgent problems since a large quantity of sewage sludge is formed by activated sludge processes. Excess sludge containing 50mM sucrose was fermented at 50 degrees C using endogenous bacteria in excess sludge, resulting in a high lactic acid production (8.45 g/L) and in an increased sludge reduction (38.2%). Conversion rate to lactic acid was up to 106.0% by standard fermentation at 50 degrees C compared to 43.8% at 30 degrees C and this phenomenon that conversion rate was higher was observed only at 50 degrees C as the fermentation at less or more than 50 degrees C had lower conversion rate than that at 50 degrees C. Lactic acid bacteria increased at 50 degrees C during 1-d fermentation whereas the number of total viable bacteria only increased slightly, indicating that lactic acid bacteria in sludge at 50 degrees C were preferentially able to utilize the sucrose for producing lactic acid. Finally, pH vibration fermentation at 50 degrees C enabled to completely consume residual sucrose in the normal fermentation, resulting in the maximum production of lactic acid. Lactate fermentation by a purely cultured lactic acid bacterium TS1 with autoclaved excess sludge containing 50mM sucrose had more than 100% of conversion rate to lactic acid, indicating that a part of sludge was converted into lactic acid during the fermentation. Our technique is useful as a facile engineering for reducing excess sludge concomitantly with producing lactic acid by lactate fermentation. PMID- 19286313 TI - Photodegradation of SF6 on polyisoprene surface: implication on elimination of toxic byproducts. AB - Photodegradation of SF(6) was performed on the surface of polyisoprene (PI) based on a brand new mechanism of "controlled release of radicals". Effective decomposition of SF(6) (60% of SF(6) was degraded in 4h) was achieved due to the highly reductive radicals (mainly allylic radicals and excited CC bond) which were generated from the photolysis of PI. No toxic fluoride was detected by FT IR. The PI irradiated for 200 h in SF(6) circumstance was examined by XPS to be doped with fluorine and sulfur. Fouling due to photoinitiated polymerization on UV lamp was avoided because the radicals were released slowly. Photolysis of SF(6) in pure argon with the presence of irradiated PI showed kinetics of pseudo first-order reaction and the degradation rate constant was 5.16 x 10(-5)s(-1). Factors which may affect the photolysis process such as introduction of O(2) and H(2)O were also examined. PMID- 19286314 TI - Immobilization of Trichoderma viride for enhanced methylene blue biosorption: batch and column studies. AB - An efficient dye biosorbent was developed by entrapping a fungus mold, Trichoderma viride, within loofa sponge (LS) matirx. Immobilization enhanced the sorption of dye by 30% at equilibrium as compared with T. viride free biomass (TVFB). The maximum dye biosorption capacity of T. viride immobilized onto loofa sponge (TVILS) and TVFB was found to be 201.52 and 155.06 mg g(-1) biomass, respectively. The kinetics of dye removal by TVILS was rapid, with 84.3% sorption within the first 30 min and equilibrium after 90 min, whereas sorption by TVFB was slower as 61.4% dye was removed in first 30 min and equilibrium was achieved in 120 min. Biosorption kinetics and equilibria followed the pseudo-second-order and Langmuir adsorption models. FTIR spectroscopy of T. viride biomass showed that amine, hydroxyl, carbonyl and amide bonds were involved in the sorption of dye. Dye desorption from dye-laden TVILS with 0.1M HCl was 99%. Regenerated TVILS was reusable without any appreciable decrease in its biosorption capacity during five repeated cycles. The dye removing capacity of TVILS in a continuous-flow column bioreactor was better than in batch-scale procedures. The study shows that TVILS has the potential of application as an efficient biosorbent for the removal of methylene blue from aqueous solutions. PMID- 19286315 TI - Treatment of spent catalyst from the nitrogenous fertilizer industry--a review of the available methods of regeneration, recovery and disposal. AB - Disposal of spent catalyst is a problem as it falls under the category of hazardous industrial waste. The recovery of metals from these catalysts is an important economic aspect as most of these catalysts are supported, usually on alumina/silica with varying percent of metal; metal concentration could vary from 2.5 to 20%. Metals like Ni, Mo, Co, Rh, Pt, Pd, etc., are widely used as a catalyst in chemical and petrochemical industries and fertilizer industries. They are generally supported on porous materials like alumina and silica through precipitation or impregnation processes. Many workers have adapted pyrometallurgy and Hydrometallurgy process for recovery of precious metals. Many workers have studied the recovery of nickel from a spent catalyst in an ammonia plant by leaching it in sulphuric acid solution (Hydrometallurgy). Ninety-nine percent of the nickel was recovered as nickel sulphate when the catalyst, having a particle size of 0.09 mm was dissolved in an 80% sulphuric acid solution for 50 min in at 70 degrees C. Many researcher have studied the extraction of metals from spent catalyst by roasting-extraction method (Pyrometallurgy). Chelating agents are the most effective extractants, which can be introduced in the soil washing fluid to enhance heavy metal extraction from contaminated soils. The advantages of chelating agents in soil cleanup include high efficiency of metal extraction, high thermodynamic stabilities of the metal complexes formed, good solubilities of the metal complexes, and low adsorption of the chelating agents on soils, But very few workers have attempted chelating agent to extract metals from spent catalyst. PMID- 19286316 TI - Converter slag-coal cinder columns for the removal of phosphorous and other pollutants. AB - A mixture of converter slag and coal cinder as adsorbent for the removal of phosphorous and other pollutants was studied in the paper. The maximum P adsorption capacity, pH of solution, contact time and initial phosphate concentration were evaluated in batch experiments for the two materials firstly. The data of P sorption were best fitted to Langumir equation, and the maximum adsorption capacities of converter slag and coal cinder were 2.417 and 0.398 mg P/g, respectively. The pH of solutions with converter slag and coal cinder changed dramatically with time and closed to 8 in 8h, and the influence of initial pH on phosphate removal by coal cinder was more significant than by converter slag. Phosphate removal rate by converter slag decreased with increase of initial phosphate concentrations. Subsequently, two flow-through columns (Column 1#, V(converter slag):V(coal cinder)=1:5; Column 2#, V(converter slag):V(coal cinder)=1:3) were operated for the removal of phosphorous and other pollutants from the effluents of a vermifilter for nearly eleven months. Results indicated the average removal efficiency of total phosphorus, dissolved phosphorus, COD and NH(4)(+)-N by Column 1# were 44%, 56%, 31% and 67%, and by Column 2# were 42%, 54%, 24% and 57%, respectively. Column 1# had higher removal efficiency for P and other pollutants. PMID- 19286317 TI - Effect of increasing nitrobenzene loading rates on the performance of anaerobic migrating blanket reactor and sequential anaerobic migrating blanket reactor/completely stirred tank reactor system. AB - A laboratory scale anaerobic migrating blanket reactor (AMBR) reactor was operated at nitrobenzene (NB) loading rates increasing from 3.33 to 66.67 g NB/m(3)day and at a constant hydraulic retention time (HRT) of 6 days to observe the effects of increasing NB concentrations on chemical oxygen demand (COD), NB removal efficiencies, bicarbonate alkalinity, volatile fatty acid (VFA) accumulation and methane gas percentage. Moreover, the effect of an aerobic completely stirred tank reactor (CSTR) reactor, following the anaerobic reactor, on treatment efficiencies was also investigated. Approximately 91-94% COD removal efficiencies were observed up to a NB loading rate of 30.00 g/m(3)day in the AMBR reactor. The COD removal efficiencies decreased from 91% to 85% at a NB loading rate of 66.67 g/m(3)day. NB removal efficiencies were approximately 100% at all NB loading rates. The maximum total gas, methane gas productions and methane percentage were found to be 4.1, 2.6l/day and 59%, respectively, at a NB loading rate of 30.00 g/m(3)day. The optimum pH values were found to be between 7.2 and 8.4 for maximum methanogenesis. The total volatile fatty acid (TVFA) concentrations in the effluent were 110 and 70 mg/l in the first and second compartments at NB loading rates as high as 66.67 and 6.67 g/m(3)day, respectively, while they were measured as zero in the effluent of the AMBR reactor. In this study, from 180 mg/l NB 66 mg/l aniline was produced in the anaerobic reactor while aniline was completely removed and transformed to 2mg/l of cathechol in the aerobic CSTR reactor. Overall COD removal efficiencies were found to be 95% and 99% for NB loading rates of 3.33 and 66.67 g/m(3)day in the sequential anaerobic AMBR/aerobic CSTR reactor system, respectively. The toxicity tests performed with Photobacterium phosphoreum (LCK 480, LUMIStox) and Daphnia magna showed that the toxicity decreased with anaerobic/aerobic sequential reactor system from the influent, anaerobic and to aerobic effluents. PMID- 19286318 TI - Utilization of ferroalumina as raw material in the production of Ordinary Portland Cement. AB - Red mud (RM), the digestion by-product of bauxite processing from "Aluminum Hellas", was dewatered by means of a filter press. The filtrand with water content from 28 to 32 wt% was named ferroalumina (FA). In order to utilize it as a raw material in the production of Ordinary Portland Cement (OPC), mixtures were prepared with limestone, sandstone and 1, 3 and 5 wt% FA addition, respectively. The design of the mixtures was based on the cement's compositional indexes LSF, AM and SM. Burnability tests showed that less than 1 wt% free lime can be obtained in all cases for firing at 1450 degrees C, except for the mixture with 1 wt% FA addition, which requires 1550 degrees C. XRD analysis and optical microscopy examination showed that FA addition did not affect the mineralogical phases of the produced clinkers. The characterisation of the produced Portland cements indicated that differences on surface area, water requirement and setting time are negligible. Compressive strength results after 28 days of curing varies from 55 to 63 MPa, which ranks the produced cements in CEM I 42.5N category. More specifically, the cements with FA addition due to their relatively high 2-day strengths (>20 MPa) can be ranked in CEM I 52.5N category. Addition of FA increases the amount of water-soluble chromium proportionally to the amount of total chromium to the mixture; however, conversion of total Cr to hexavalent Cr remains practically constant, in the range of 32-35 wt%. The results indicate that FA can be used as raw material in the production of OPC up to 5 wt% according to the chemical composition of the other raw materials. PMID- 19286319 TI - Optimization of process parameters for the extraction of chromium (VI) by emulsion liquid membrane using response surface methodology. AB - The emulsion liquid membrane technique was used for the extraction of hexavalent chromium ions from aqueous solution of waste sodium dichromate recovered from the pharmaceutical industry wastewater. The liquid membrane used was composed of kerosene oil as the solvent, Span-80 as the surfactant and potassium hydroxide as internal reagent. Trioctyl amine and Aliquat-336 were used as carriers. The emulsion stability was carried out at different surfactant concentration, agitation speed and emulsification time. Statistical experimental design was applied for the optimization of process parameters for the extraction of chromium by emulsion liquid membrane. The effects of process parameters namely, agitation speed, membrane to emulsion (M/E) ratio and carrier concentration on the extraction of chromium were optimized using a response surface method. The optimum conditions for the extraction of chromium (VI) using response surface methodology for Trioctyl amine were: agitation speed--201.369rpm, M/E ratio- 0.5887% (v/v) and carrier concentration--4.0932% (v/v) and for Aliquat-336: agitation speed--202.097 rpm, M/E ratio--0.5873% (v/v) and carrier concentration -3.9211% (v/v). At the optimized condition the maximum chromium extraction was found to be 89.2% and 96.15% using Trioctyl amine and Aliquat-336, respectively. PMID- 19286320 TI - Molecular profiles of Venezuelan isolates of Trypanosoma sp. by random amplified polymorphic DNA method. AB - Nine Trypanosoma sp. Venezuelan isolates, initially presumed to be T. evansi, were collected from three different hosts, capybara (Apure state), horse (Apure state) and donkey (Guarico state) and compared by the random amplification polymorphic DNA technique (RAPD). Thirty-one to 46 reproducible fragments were obtained with 12 of the 40 primers that were used. Most of the primers detected molecular profiles with few polymorphisms between the seven horse, capybara and donkey isolates. Quantitative analyses of the RAPD profiles of these isolates revealed a high degree of genetic conservation with similarity coefficients between 85.7% and 98.5%. Ten of the primers generated polymorphic RAPD profiles with two of the three Trypanosoma sp. horse isolates, namely TeAp-N/D1 and TeGu N/D1. The similarity coefficient between these two isolates and the rest, ranged from 57.9% to 68.4% and the corresponding dendrogram clustered TeAp-N/D1 and Te Gu-N/D1 in a genetically distinct group. PMID- 19286321 TI - Histopathological changes of Korea cultured olive flounder, Paralichthys olivaceus due to scuticociliatosis caused by histophagous scuticociliate, Philasterides dicentrarachi. AB - Recently, mass mortality due to histophagous scuticociliate, Philasterides dicentrarachi in olive flounder, Paralichthys olivaceus occurred in aquaculture farms of Jeju Island, South Korea. We have artificially induced infection through immersion on day 12, the density of ciliates slowly decreased in the experimental tanks; on day 28 to 2300cells/ml in 3cm group and 1800cells/ml in 5cm group. The survival rate of P. olivaceus on days 24 and 28 of infection in the 3cm group was decreased to 4 and 6, respectively; however, no mortality was recorded in the 5cm group. In the control group 100% survival in the chosen period of both groups occurred except in the 3cm group whereas on days 24 and 28 the death rate was 1 and 2, respectively. Many ciliates with ingested red blood cells were observed in the gills, skeletal muscle, skin, fins and brains of infected flounder. The parasites were also observed in the lamina propria of the digestive tract, pharynx, peritoneal fluid, cerebrospinal fluid and cornea. Histopathological and inflammatory changes were characterized by swelling and generalized erythema and muscle tissue necrosis. Affected fish showed varying levels of tissue damage including severe epidermal and dermal necrotic lesions. Loss of scales, appearance of bleached spots that coalesced to form brownish patches, haemorrhagic and severe dermal necrotic lesions were the major clinico pathological manifestations. The parasite was also seen distributed extensively in the entire brain causing widespread nerve necrosis after a prolonged period of infection. These results appear to support the hypothesis that the P. dicentrarchi may be penetrating via the gills or the skin, and then travel via the bloodstream to other parts of the body, including the body cavity. PMID- 19286322 TI - The efficiency of avermectins (abamectin, doramectin and ivermectin) in the control of Boophilus microplus, in artificially infested bovines kept in field conditions. AB - Tests were performed on artificially infested bovines, kept in field conditions, to assess the efficiency of avermectins (abamectin, doramectin and ivermectin) on Boophilus microplus (Canestrini, 1887). This assessment was carried out on 40 bovines, in the Paraiba Valley, in the State of Sao Paulo, Brazil. These bovines were distributed into four groups (abamectin, doramectin, ivermectin and a control group), after artificial infestation with some 4000 larvae per animal on days -21, -14, -7, -1, 7 and 14. The animals from the treated groups were subcutaneously injected with the commercial avermectins, at a dose of 200microg/kg of live weight (1mL/50kg). The best results were shown by the group treated with doramectin, both in relation to the reduction of female count (an average of 85.92% between 3rd and 28th day after treatment) and also in the reduction of oviposition among the females collected from the bovines after treatment and then kept in the laboratory (an average of 83.51%). None of the avermectins proved to be efficient in inhibiting the hatching of the larvae. PMID- 19286323 TI - Propetamphos resistance in Rhipicephalus bursa (Acari, Ixodidae). AB - Hard tick infestation is an important problem in both traditional and industrial animal husbandry as well as in veterinary and public health. Several acaricides have been used to control tick infestation in Iran. Poor control has been attributed to acaricide resistance in tick populations although this has not been demonstrated experimentally. In this study, susceptibility status to propetamphos (Blotic) of 2-3-week-old larvae of Rhipicephalus bursa, one of the most common tick species in Iran, was evaluated using the FAO recommended larval packet test (LPT) method with some modifications. Log dosage probit analysis of propetamphos dose-response regression lines produced a resistance ratio based on LC(50) in the most resistant strain of approximately 103-fold. Biochemical assays suggested that the tick strains contained multiple insecticide resistance mechanisms involving elevation of esterases, GSTs, MFOs and alteration of acetylcholinesterase. In light of this data, practical implications for pest and pesticide resistance management strategies are discussed. PMID- 19286324 TI - Pseudomonas pyocyanin inhibits wound repair by inducing premature cellular senescence: role for p38 mitogen-activated protein kinase. AB - Pseudomonas aeruginosa is an important nosocomial pathogen of burn wounds. Pyocyanin, a virulence factor produced by the bacterium, induces persistent intracellular oxidative stress and premature senescence in mammalian cells. Our aims were to evaluate pyocyanin levels in infected wound dressings and the potential of the toxin to influence wound repair. Surgical dressings from infected burn patients were examined for pyocyanin and normal primary human diploid fibroblasts (HDFs) were treated with comparable concentrations and their replicative ability examined. Pyocyanin was detected in the exudate of infected wound dressings in amounts up to 5.3 microg/g (mean: 2.0+/-2.3 microg/g). HDFs exposed to pyocyanin (1-50 microM; 0.2-10.5 microg/ml) underwent growth arrest at all concentrations and developed morphological characteristics associated with cellular senescence, including expression of senescence-associated beta galactosidase. Using an in vitro wound repair model, a single exposure to pyocyanin inhibited wound repair in a concentration-dependent manner. Prior treatment with a specific p38(MAPK) inhibitor allowed cells to maintain their replicative ability and pre-senescent morphology indicating pyocyanin operates through the Erk/p38(MAPK) senescence pathway. These data support the hypothesis that bacterial virulence factors capable of inducing persistent low-level oxidative stress play a pivotal role in modulating the tissue repair response to infection by inducing premature cellular senescence. PMID- 19286325 TI - The antimicrobial effect of acetic acid--an alternative to common local antiseptics? AB - Acetic acid has been commonly used in medicine for more than 6000 years for the disinfection of wounds and especially as an antiseptic agent in the treatment and prophylaxis of the plague. The main goal of this study was to prove the suitability of acetic acid, in low concentration of 3%, as a local antiseptic agent, especially for use in salvage procedures in problematic infections caused by organisms such as Proteus vulgaris, Acinetobacter baumannii or Pseudomonas aeruginosa. This study was designed to compare the in vitro antimicrobial effect of acetic acid with those of common local antiseptics such as povidone-iodine 11% (Betaisodona), polyhexanide 0.04% (Lavasept), mafenide 5% and chlohexidine gluconate 1.5% cetrimide 15% (Hibicet). Former studies suggest the bactericidal effect of acetic acid, but these data are very heterogeneous; therefore, a standardised in vitro study was conducted. To cover the typical bacterial spectrum of a burn unit, the following Gram-negative and Gram-positive bacterial strains were tested: Escherichia coli, P. vulgaris, P. aeruginosa, A. baumannii, Enterococcus faecalis, Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus (MRSA) and beta-haemolytic Streptococcus group A and B. The tests showed excellent bactericidal effect of acetic acid, particularly with problematic Gram-negative bacteria such as P. vulgaris, P. aeruginosa and A. baumannii. The microbiological spectrum of acetic acid is wide, even when tested at a low concentration of 3%. In comparison to our currently used antiseptic solutions, it showed similar - in some bacteria, even better - bactericidal properties. An evaluation of the clinical value of topical application of acetic acid is currently underway. It can be concluded that acetic acid in a concentration of 3% has excellent bactericidal effect and, therefore, seems to be suitable as a local antiseptic agent, but further clinical studies are necessary. PMID- 19286326 TI - Safety preparedness of urban community for New Year fireworks in Tehran. AB - Fireworks are used worldwide during national and cultural celebrations. Iranians celebrate the last Wednesday of the Persian year (ILWEF) as an ancient custom. The aim of this study was to determine safety preparedness of inhabitation in Tehran, the capital city of Iran, for 2007 ILWEF fireworks. In this study, interviewers referred to randomly selected addresses after the end of ILWEF. They asked responders about source of information on how to use firework items and having education of safe use of fireworks. Also people were asked about having first aid kits and fire extinguisher at home. The study included 2475 families. To learn how to use fireworks, 1.3% had read brochures; the remainder of responders obtained their information from miscellaneous sources with unknown validity. Only 1.1% of them have had some sort of education on the safe use of fireworks. According to responders, 98.4% did not have any information about the safety standards of fireworks. We conclude that Tehranian households were not sufficiently prepared to guarantee a safe festival. It seems safety preparedness of the community to prevent firework related fires and injuries, by relevant education and promotion of fire safety equipments, needs to be reconsidered in Tehran. Considering the impaired safety preparedness of community for fireworks festival, individuals should attend public fireworks displays conducted by professionals rather than engage in personal use of fireworks. Restriction of the use of fireworks and banning firecrackers lacking appropriate insertions on safety instructions are other solutions to be considered. PMID- 19286327 TI - Nodular lymphoid hyperplasia and histologic changes mimicking celiac disease, collagenous sprue, and lymphocytic colitis in a patient with selective IgA deficiency. AB - Selective IgA deficiency is the most common primary immunoglobulin deficiency. The clinical manifestations of selective IgA deficiency, including gastrointestinal (GI) complications, are rare and typically milder than those seen with common variable immunodeficiency or X-linked agammaglobulinemia. We present a rare case of selective IgA deficiency that shows a number of interesting histological features in the GI tract, including diffuse nodular lymphoid hyperplasia involving the entire small and large intestine, celiac disease-like and collagenous sprue-like changes in the small intestine, as well as lymphocytic colitis pattern. However, this patient had no particular GI symptoms suggestive of celiac sprue or microscopic colitis. These findings suggest that the GI tract in patients with selective IgA deficiency can show peculiar histologic changes that mimic celiac disease, collagenous sprue, or lymphocytic colitis, which may be a pattern of injury related to infection or immunoglobulin immunodeficiency-associated autoimmune phenomena. PMID- 19286328 TI - Correlation of point B and lymph node dose in 3D-planned high-dose-rate cervical cancer brachytherapy. AB - PURPOSE: To compare high dose rate (HDR) point B to pelvic lymph node dose using three-dimensional-planned brachytherapy for cervical cancer. METHODS AND MATERIALS: Patients with FIGO Stage IB-IIIB cervical cancer received 70 tandem HDR applications using CT-based treatment planning. The obturator, external, and internal iliac lymph nodes (LN) were contoured. Per fraction (PF) and combined fraction (CF) right (R), left (L), and bilateral (Bil) nodal doses were analyzed. Point B dose was compared with LN dose-volume histogram (DVH) parameters by paired t test and Pearson correlation coefficients. RESULTS: Mean PF and CF doses to point B were R 1.40 Gy +/- 0.14 (CF: 7 Gy), L 1.43 +/- 0.15 (CF: 7.15 Gy), and Bil 1.41 +/- 0.15 (CF: 7.05 Gy). The correlation coefficients between point B and the D100, D90, D50, D2cc, D1cc, and D0.1cc LN were all less than 0.7. Only the D2cc to the obturator and the D0.1cc to the external iliac nodes were not significantly different from the point B dose. Significant differences between R and L nodal DVHs were seen, likely related to tandem deviation from irregular tumor anatomy. CONCLUSIONS: With HDR brachytherapy for cervical cancer, per fraction nodal dose approximates a dose equivalent to teletherapy. Point B is a poor surrogate for dose to specific nodal groups. Three-dimensional defined nodal contours during brachytherapy provide a more accurate reflection of delivered dose and should be part of comprehensive planning of the total dose to the pelvic nodes, particularly when there is evidence of pathologic involvement. PMID- 19286329 TI - Synergistic effects of hemoglobin and tumor perfusion on tumor control and survival in cervical cancer. AB - PURPOSE: The tumor oxygenation status is likely influenced by two major factors: local tumor blood supply (tumor perfusion) and its systemic oxygen carrier, hemoglobin (Hgb). Each has been independently shown to affect the radiotherapy (RT) outcome in cervical cancer. This study assessed the effect of local tumor perfusion, systemic Hgb levels, and their combination on the treatment outcome in cervical cancer. METHODS AND MATERIALS: A total of 88 patients with cervical cancer, Stage IB2-IVA, who were treated with RT/chemotherapy, underwent serial dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) before RT, at 20 22 Gy, and at 45-50 Gy. The DCE-MRI perfusion parameters, mean and lowest 10th percentile of the signal intensity distribution in the tumor pixels, and the Hgb levels, including pre-RT, nadir, and mean Hgb (average of weekly Hgb during RT), were correlated with local control and disease-specific survival. The median follow-up was 4.6 years. RESULTS: Local recurrence predominated in the group with both a low mean Hgb (<11.2 g/dL) and low perfusion (lowest 10th percentile of signal intensity <2.0 at 20-22 Gy), with a 5-year local control rate of 60% vs. 90% for all other groups (p = .001) and a disease-specific survival rate of 41% vs. 72% (p = .008), respectively. In the group with both high mean Hgb and high perfusion, the 5-year local control rate and disease-specific survival rate was 100% and 78%, respectively. CONCLUSION: These results suggest that the compounded effects of Hgb level and tumor perfusion during RT influence the radioresponsiveness and survival in cervical cancer patients. The outcome was worst when both were impaired. The management of Hgb may be particularly important in patients with low tumor perfusion. PMID- 19286330 TI - Predicting the risk of pelvic node involvement among men with prostate cancer in the contemporary era. AB - PURPOSE: The "Roach formula" for the risk of pelvic lymph node metastases [(2/3) ( *) PSA + (Gleason score - 6) ( *) 10] was developed in the early prostate specific antigen (PSA) era. We examined the accuracy of this formula in contemporary patients. METHODS: We included men in the Surveillance, Epidemiology, and End Results Registry with a diagnosis of clinical T1c-T4 prostate cancer in 2004 who had a surgical lymph node evaluation, Gleason score (typically from prostatectomy), and baseline PSA level (n = 9,387). Expected and observed rates of node positivity were compared. RESULTS: Ninety-eight percent were clinical T1c/T2, and 97% underwent prostatectomy. Overall, 309 patients (3.29%) had positive lymph nodes. Roach scores overestimated the actual rate of positive lymph nodes in the derivation set by 16-fold for patients with Roach score less than or equal to 10%, by 7-fold for scores greater than 10-20%, and by approximately 2.5-fold for scores greater than 20%. Applying these adjustment factors to Roach scores in the validation data set yielded accurate predictions of risk. For those with Roach score less than or equal to 10%, adjusted expected risk was 0.2% and observed risk was 0.2%. For Roach score greater than 10-20%, adjusted expected risk was 2.0% and observed risk was 2.1%. For Roach score greater than 20-30%, adjusted expected risk was 9.7% and observed risk was 6.5%. For Roach score greater than 30-40%, adjusted expected risk was 13.9% and observed risk was 13.9%. CONCLUSION: Applied to contemporary patients with mainly T1c/T2 disease, the Roach formula appears to overestimate pelvic lymph node risk. The adjustment factors presented here should be validated by using biopsy Gleason scores and extended lymphadenectomies. PMID- 19286331 TI - Dose de-escalation with gamma knife radiosurgery in the treatment of choroidal melanoma. AB - PURPOSE: Single-fraction targeted radiation therapy delivered by the Leksell Gamma Knife system is a minimally invasive treatment option for choroidal melanoma that has been used as an alternative to enucleation, proton beam therapy, or brachytherapy. Previously reported Gamma Knife series involved the treatment of choroidal melanomas with a dose of 40 to 50 Gy at the tumor margin. We report our institutional experience using a significantly lower dose. METHODS AND MATERIALS: Fourteen patients with choroidal melanoma were treated with the Leksell Gamma Knife at our institution over a 7-year period. The treatment and clinical data were analyzed in a retrospective fashion after a mean follow-up of 32.2 months. RESULTS: The mean dose to the tumor margin was 22.2 +/- 2.4 Gy (range, 20- 25 Gy). Mean treated tumor volume was 1.1 +/- 1.2 cc. Local control was achieved in 13 cases (93%). In 1 patient both intraocular spread and distant metastatic disease developed after treatment. Visual function of the affected eye was preserved in 5 patients (36%) at latest follow-up, in 9 patients (64%) visual loss ensued. Mild to moderate radiation toxicity developed in 8 patients. CONCLUSIONS: Choroidal melanoma can be safely and effectively treated using Leksell Gamma Knife stereotactic radiosurgery with a marginal dose of less than 25 Gy. PMID- 19286332 TI - Thyroid function in women after multimodal treatment for breast cancer stage II/III: comparison with controls from a population sample. AB - PURPOSE: A possible association between thyroid diseases (TD) and breast cancer (BC) has been debated. We examined prevalence and development of TD in women after multimodal treatment for Stage II/III BC compared with women from a general population. Secondarily, we explored the impact of two different radiotherapy (RT) techniques (standardized field arrangements vs. computed tomography [CT] based dose planning) on TD in BC patients examined 35-120 months after primary BC treatment. METHODS AND MATERIALS: A total of 403 BC patients completed a questionnaire about TD and had blood samples taken for analyses of thyroid function. All had undergone postoperative RT with or without (2%) adjuvant systemic treatment. The results in the BC patients were compared with a cancer free, age-matched control group from a general population (CGr). RESULTS: There was higher prevalence of self-reported hypothyroidism in the BC patients as compared with the CGr (18% vs. 6%, p < 0.001). The raised prevalence was predominantly due to a substantial increase in the development of hypothyroidism after BC diagnosis, whereas the prevalence of hypothyroidism before BC diagnosis was similar to that observed in the CGr. Patients treated with CT-based RT showed a trend for increased post-BC development of hypothyroidism as compared with those treated with standardized field arrangements (p = 0.08). CONCLUSIONS: Hypothyroidism is significantly increased in women after multimodal treatment for Stage II/III BC. Radiation to the thyroid gland may be a contributing factor. BC patients should be routinely screened for hypothyroidism. PMID- 19286333 TI - [MiniArc: preliminary prospective study on 72 cases]. AB - OBJECTIVES: To present a new mini-invasive tape requiring only a single incision, to assess the complications and short-term results. MATERIALS AND METHODS: Prospective study of 72 patients with stress urinary incontinence. Were selected for the study patients with stress urinary incontinence with or without intrinsic sphincter deficiency and mixed urinary incontinence with a predominance of leaks to the effort. Patients were evaluated at 2 months of the intervention. RESULTS: Isolated stress urinary incontinence for 58 patients, mixed incontinence for 14 and intrinsic sphincter deficiency for five. The complications per procedure revealed one bleeding over 100ml, one malaise vagal and a failure of the procedure. At 2 months, the authors found: two urgenturies de novo, one de novo micturition disorder, one erosion, one urinary tract infection and four lateral vaginal bands. The early objective results are overall 75.7% of dry patients (79.6% for pure SUI, 66.6% for IUE with ISD and 61.5% for mixed incontinence). The overall failure rate was 2.9%. CONCLUSION: The results of this simplified tape are worse than those of TVT and transobturator tapes. PMID- 19286334 TI - [Perinatal health: situation in France and in the other members of the European Union]. PMID- 19286335 TI - TCF7L2 is associated with type 2 diabetes in nonobese individuals from Tunisia. AB - The transcription factor 7-like 2 (TCF7L2) rs7903146 T allele was associated with type 2 diabetes (T2D) in most populations worldwide. In individuals of European descent, the association with T2D was recently found to be modulated by obesity status. However, further studies are necessary to clarify if whether interaction exists among subjects of non-European descent. In the present study, we analyzed the association of rs7903146 with T2D in 90 nonobese (Body Mass Index [BMI] <25kg/m(2)), 171 overweight (25<=BMI<30kg/m(2)) et 98 obese (BMI>=30kg/m(2)) individuals from Tunisia. The T allele was nominally associated with T2D in nonobese subjects (Odds Ratio [OR]=3.24 [1.10-9.53], P=0.021) whereas no effect was detected in overweight (P=0.3) and obese (P=0.22) individuals. Consequently, the same risk allele decreased susceptibility to obesity in T2D subjects (OR=0.47 [0.23-0.94], P=0.029) but not in normoglycemic controls (P=0.44). When analyzed all together, no allelic association was observed with T2D (P=0.20) whereas an artefactual association with decreased obesity (0.59 [0.38-0.90], P=0.013) was detected. As in Europeans, TCF7L2 is therefore not a risk factor for obesity in Tunisians, but its effect on T2D risk is modulated by obesity. In conclusion, the TCF7L2 rs7903146 T allele is nominally associated with T2D susceptibility in nonobese individuals from Tunisia. PMID- 19286336 TI - Antioxidant vitamin status during pregnancy in relation to cognitive development in the first two years of life. AB - OBJECTIVE: To investigate the correlation of the antioxidant vitamins status (vitamins A, E and C) during pregnancy and the intellectual development of early childhood. METHOD: A total of 150 paired maternal-neonatal subjects were recruited into the present study. The serum concentrations of antioxidant vitamins (vitamins A, E and C) in maternal blood and cord blood after delivery were determined by high performance liquid chromatography and the intellectual development was evaluated by Gesell Development Schedules (GDS) at two-years-old. RESULT: Children with higher cord serum vitamin E level showed higher scores of motor, adaptive domain and average compared to children with lower cord serum vitamin E level (p<0.01 or 0.05), respectively. Cord serum vitamin A level had significant positive correlation with effect on motor DQs (beta=4.227, p<0.05), and vitamin E level in cord blood showed a positive relation with motor DQ and average DQ (beta=0.329 and 0.1875, respectively, p<0.05) in multiple linear regression model. The language and social DQs were influenced by placental vitamin E transport rate (beta=3.1968 and 3.0194, respectively, p<0.05). The placental transport rate of vitamin E also was a protective factor for the prevalence of motor behavior developmental delay [OR: 0.118, 95% confident interval (95% CI), 0.018-0.765, p=0.0251], personal and social behavior developmental delay (OR: 0.052, 95% CI: 0.004-0.610, p=0.0185) and average developmental delay (OR: 0.041, 95% CI: 0.003-0.642, p=0.0229) in logistic multiple regression model. CONCLUSION: Data suggested that vitamin A, E status and vitamin E transfer rate at delivery had beneficial influence on children's cognitive and behavior development quotients. PMID- 19286337 TI - Estimating stature from percutaneous length of tibia and ulna in Indo-Mauritian population. AB - Stature estimation from percutaneous body measurements forms part of forensic anthropological analysis for the purpose of identification. This study is aimed at modeling the stature on the basis of percutaneous tibial and/or ulnar length in human subjects comprising of Indo-Mauritian population. The study was conducted in the Department of Forensic Medicine and Toxicology, SSR Medical College, Mauritius on 180 young and healthy students comprising of 90 males and 90 females in the age group ranging from 18 to 28 years. The measurements were taken by using standard anthropometric instruments. It is remarked that tibial as well as ulnar length show a linear relationship with the stature, where stature is normally distributed. However, it is required to transform the measurements on stature to obtain appropriate regression equations. Moreover, since ulna and tibia are significantly correlated, it is recommended to use the sum of the ulnar and tibial length to estimate stature, in case both are available for an individual. Our regression models are sufficiently validated and highly efficient. PMID- 19286338 TI - Improved SNR in linear reordered 2D bSSFP imaging using variable flip angles. AB - This article presents a variable flip-angle approach for balanced steady-state free precession (bSSFP) imaging, which allows increases in signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) while keeping specific absorption rate (SAR) constant or reduces SAR for given CNR and SNR. The gain in SNR is achieved by utilizing the higher signal in the transient phase. Flip-angle variation during the echo train is realized using a trigonometric function with M steps (ramp length). Variation is combined with a linear k-space reordering such that outer parts of k-space are sampled using a lower flip angle alpha(min), while the central part of k-space is acquired with a higher flip angle alpha(max). No additional preparation or dummy cycles are applied prior to data acquisition. Several variation schemes with different starting flip angles alpha(min) and ramp length M are considered. For example, using alpha(min)=1 degrees and M=96, alpha(max) can be set to 47 degrees without exceeding SAR limits at 3 T and gaining up to 50% in SNR, while, conventionally, alpha=34 degrees is the maximal possible flip angle. Resolution seems unaffected in volunteer imaging. In all cases, no transient artifacts due to flip-angle variation were observed. This article demonstrates the use of flip-angle variations in bSSFP to increase SNR and CNR while keeping SAR constant, which is especially important at higher field strengths. Flip-angle variation can also be combined with other methods such as parallel imaging techniques for further SAR reduction. PMID- 19286339 TI - Development and validation of a reversed-phase HPLC method for the simultaneous analysis of simvastatin and tocotrienols in combined dosage forms. AB - A RP-HPLC method for the simultaneous analysis of tocotrienol isoforms (TRF) and simvastatin (SIM) in SIM-TRF nanoparticles (NPs) was developed. Analytes were monitored by UV detection at 238 and 295 nm for SIM and TRF, respectively, using a gradient methanol/water elution. Calibration curves for TRF and SIM were linear over concentration range of 20-80 microg/mL and 1-10 microg/mL with correlation coefficients 0.9990 and 0.9991, respectively. The recovery of TRF and SIM from the NPs was in the range from 97.35 to 102.19% and from 92.71 to 104.35%, respectively. This developed method was successfully employed in quantifying both drugs in NPs for future use in cancer therapy. PMID- 19286340 TI - Production of uremic toxin methylguanidine from creatinine via creatol on activated carbon. AB - The mixture of creatinine, activated charcoal and water was stirred. As a result the conversion of creatinine into two products was observed. (1)H, (13)C NMR and HMBC spectra were recorded and the chemical shifts assigned. Two uremic toxins: creatol and N-methylguanidine were identified. To interpret the NMR data obtained, the optimum structure of creatol, which can exist in the forms of seven tautomers, has been calculated using the DFT B3LYP/6-311G(2d,p) method. The influence of the solvent was described by the polarizable continuum model (PCM). The calculated energy of the most energetically stable tautomeric form A is lower by 12.2, 16.9, 33.8, 81.5, 106.3, 130.4kJ/mol in water than that of the tautomers B-G, respectively, which suggests that the A form of creatol should prevail in solution. In DMSO, the calculated energy of the most energetically stable tautomeric form A is lower than that of both D and B and the remaining tautomeric forms (C, E-G) are less energetically stable. Subsequently, we sought the correlations between the experimental and the calculated chemical shifts of protons and carbons-13 for the forms -A, B (in water) and A, B, D (in DMSO) - of creatol. The population of the A tautomer is predominant in both H(2)O and DMSO. We have also recorded the spectra of creatol and N-methylguanidine at different pH. Our data are complete enough to be used in the analysis of body fluids. PMID- 19286341 TI - Validity aspects of the patient feedback questionnaire on consultation skills (PFC), a promising learning instrument in medical education. AB - OBJECTIVE: A focus on the communicator competency is considered to be an important requirement to help physicians to acquire consultation skills. A feedback questionnaire, in which patients assess consultation skills might be a useful learning tool. An existing questionnaire on patient perception of patient centeredness (PPPC) was adapted to cover the 'communicator' items in the competency profile. We assessed the face and content validity, the construct validity and the internal consistency of this new patient feedback on consultation skills (PFC) questionnaire. METHODS: We assessed the face validity of the PFC by interviewing patients and general practice trainees (GPTs) during the developmental process. The content validity was determined by experts (n=10). First-year GPTs (23) collected 222 PFCs, from which the data were used to assess the construct validity (factor analysis), internal consistency, response rates and ceiling effects. RESULTS: The PFC adequately covers the corresponding 'communicator' competency (face and content validity). Factor analysis showed a one-dimensional construct. The internal consistency was high (Cronbach's alpha 0.89). For the single items, the response rate varied from 89.2% to 100%; the maximum score (ceiling effect) varied from 45.5% to 89.2%. CONCLUSION: The PFC appears to be a valid, internally consistent instrument. PRACTICAL IMPLICATIONS: The PFC may be a valuable learning tool with which GPTs, other physicians and medical students can acquire feedback from patients regarding their consultation skills. PMID- 19286342 TI - Individuals' responses to global CHD risk: a focus group study. AB - OBJECTIVE: To explore how individuals respond to global coronary heart disease (CHD) risk and use it in combination with treatment information to make decisions to initiate and maintain risk reducing strategies. METHODS: We conducted four focus groups of individuals at risk for CHD (n=29), purposively sampling individuals with each of several risk factors. Two reviewers coded verbatim transcripts and arbitrated differences, using ATLAS.ti 5.2 to facilitate analysis. RESULTS: Participants generally regarded the concept of global CHD risk as useful and motivating, although had questions about its precision and comprehensiveness. They identified several additional influential factors in decision-making (e.g. achievable risk, the quickness and self-evidence of results) and generally preferred lifestyle changes to medications (although most would accept medications under certain circumstances). They also noted the importance of participating in decision-making. CONCLUSION: Our results underscore the motivating potential of global CHD risk and the importance of patient participation in decision-making. PRACTICE IMPLICATIONS: Global CHD risk is a useful adjunct to CHD prevention and can be presented in ways, and with information, that might improve CHD outcomes. PMID- 19286343 TI - The use of illustration to improve older adults' comprehension of health-related information: is it helpful? AB - OBJECTIVE: To examine whether explanatory illustrations can improve older adults' comprehension of written health information. METHODS: Six short health-related texts were selected from websites and pamphlets. Young and older adults were randomly assigned to read health-related texts alone or texts accompanied by explanatory illustrations. Eye movements were recorded while reading. Word recognition, text comprehension, and comprehension of the illustrations were assessed after reading. RESULTS: Older adults performed as well as or better than young adults on the word recognition and text comprehension measures. However, older adults performed less well than young adults on the illustration comprehension measures. Analysis of readers' eye movements showed that older adults spent more time reading illustration-related phrases and fixating on the illustrations than did young adults, yet had poorer comprehension of the illustrations. CONCLUSION: Older adults might not benefit from text illustrations because illustrations can be difficult to integrate with the text. PRACTICE IMPLICATIONS: Health practitioners should not assume that illustrations will increase older adults' comprehension of health information. PMID- 19286344 TI - How a hunger strike taught me to listen. PMID- 19286345 TI - Artifactual hypoglycemia in a patient with alcoholic hepatitis and hyperleukocytosis. PMID- 19286346 TI - Pulmonary artery sarcoma, a paradigm of orphan thoracic oncology. PMID- 19286347 TI - Recognition of emotional and nonemotional facial expressions: a comparison between Williams syndrome and autism. AB - The aim of our study was to compare two neurodevelopmental disorders (Williams syndrome and autism) in terms of the ability to recognize emotional and nonemotional facial expressions. The comparison of these two disorders is particularly relevant to the investigation of face processing and should contribute to a better understanding of social behaviour and social cognition. Twelve participants with WS (from 6;1 to 15 years) and twelve participants with autism (from 4;9 to 8 years) were matched on verbal mental age. Their performances were compared with those of twelve typically developing controls matched on verbal mental age (from 3;1 to 9;2). A set of five tasks assessing different dimensions of emotional and nonemotional facial recognition were administered. Results indicated that recognition of emotional facial expressions is more impaired in Williams syndrome than in autism. Our study comparing Williams syndrome and autism over a small age range highlighted two distinct profiles which call into question the relationships between social behaviour/cognition and emotion perception. PMID- 19286348 TI - Analysis of T cell receptor beta chains that combine with dominant conserved TRAV5D-4*04 anti-insulin B:9-23 alpha chains. AB - OBJECTIVE: The objective of this study was to define the spectrum of TCR beta chains permissive for T cells with alpha chains containing the conserved TRAV5D 4*04 sequence to target the insulin B:9-23 peptide, a major epitope for initiation of diabetes in the NOD mouse. MATERIALS AND METHODS: We produced T cell hybridomas from mice with single T cell receptors (BDC12-4.1 TCR alpha(+)beta(+) double transgenic mice and BDC12-4.4 TCR alpha(+)beta(+) double retrogenic mice) or from mice with only the corresponding alpha chains transgene or retrogene and multiple endogenous TCR beta chains. RESULTS: Hybridomas with the complete BDC12-4.1 and BDC12-4.4 T cell receptors, despite having markedly different TCR beta chains, responded to similar B:9-23 peptides. Approximately 1% of the hybridomas from mice with the fixed TRAV5D-4*04 alpha chains and multiple endogenous beta chains responded to B:9-23 peptides while the majority of hybridomas with different beta chains did not respond. There was no apparent conservation of TCR beta chain sequences in the responding hybridomas. CONCLUSIONS: Approximately 1% of hybridomas utilizing different TCR beta chains paired with the conserved TRAV5D-4*04 containing alpha chains respond to insulin peptide B:9-23. Therefore, TCR beta chain sequences make an important contribution to insulin B:9-23 peptide recognition but multiple beta chain sequences are permissive for recognition. PMID- 19286349 TI - Effects of nutritional intervention on body weight and body composition of obese psychiatric patients taking olanzapine. AB - OBJECTIVE: Weight gain is an established side effect of atypical antipsychotics in patients with severe mental illness (SMI). Previous studies have shown positive effects of nutritional interventions in weight loss. The purpose of this study was to investigate the effects of a nutritional intervention on the body weight and body composition of patients with SMI taking olanzapine in Greece. METHODS: Eighty-two patients with SMI treated with olanzapine (22 men, 60 women) and 58 healthy controls (12 men, 46 women) were followed for 3 mo. All patients with SMI were obese, with an average body mass index of 33.12 +/- 0.74 kg/m(2) and body weight of 94.61 +/- 2.50 kg. A nutritional program was designed for each participant based on anthropometric characteristics, health profile, and dietary needs. Pre- and postintervention anthropometric and body composition measurements were performed. RESULTS: Significant weight loss and fat loss were found in the healthy controls and patients with SMI from baseline to 3 mo (P < 0.05). However, the patients with SMI had a less significant decrease in waist circumference (P < 0.05) compared with healthy controls. The healthy male controls and male patients with SMI demonstrated greater decreases in body weight and waist circumference compared with female participants (P < 0.05). CONCLUSION: Patients with SMI appear to respond effectively to a nutritional program demonstrating significant decreases in body weight and body composition despite the use of olanzapine. Because gender differences may exist in weight loss, it is possible that gender should be taken into account for a more appropriate treatment of obesity in this population. PMID- 19286350 TI - Glutamine attenuates lipopolysaccharide-induced acute lung injury. AB - OBJECTIVES: It has been reported that glutamine (GLN) can attenuate acute lung injury after sepsis. GLN is also thought to be a precursor of glutathione (GSH) synthesis. Using the GSH synthesis blocker, L-buthionine-(S,R)-sulfoximine (BSO), we investigated the role of GSH synthesis in the protective effect of GLN on acute lung injury. METHODS: In this study, we used an acute lung injury model induced by intratracheal injection of lipopolysaccharide (1 mg mL(-1) kg(-1)). GLN (0.75 g/kg, intravenous) and BSO (2 mmol/kg, intraperitoneal) were administrated simultaneously. At 2 and 18 h after the injections, the rats were sacrificed by right ventricular puncture and bronchoalveolar lavage was done. The lower right lung was excised for histologic examination. Total protein concentration and total cell and neutrophil counts in the bronchoalveolar lavage fluid were determined. CD11b expression in the blood was determined by flow cytometry. We also analyzed myeloperoxidase activity, and GSH and interleukin-8 levels in lung tissues. RESULTS: GLN supplementation reduced the total protein concentration and total cell and neutrophils counts in bronchoalveolar lavage fluid after lipopolysaccharide challenge. GLN enhanced GSH synthesis and attenuated interleukin-8 release and myeloperoxidase activity in lung tissues. GLN also decreased CD11b expression in blood neutrophils and prevented lung histologic changes. BSO abolished the effects of GLN and attenuated its protection on acute lung injury. CONCLUSION: These results indicate that GLN could prevent neutrophil recruitment and infiltration, protect the alveolar barrier, and attenuate inflammatory injury during sepsis. This effect may be related to enhanced GSH synthesis. PMID- 19286351 TI - Recruitment of motor cortex inhibition differentiates between generalized and focal epilepsy. AB - Intracortical disinhibitory mechanisms play a crucial role in epilepsy. Therefore, the recruitment of motor cortical excitability was evaluated to distinct between focal and generalized epileptic syndromes. Twenty-five untreated patients with epilepsy and 20 controls were enrolled. Classification into focal (FE, n=10) or idiopathic generalized (IGE, n=15) epilepsy was based on seizure semiology, EEG and MRI. The recruitment of motor cortical inhibition and facilitation was measured by varying the stimulus intensity (SI) of the first conditioning stimulus in a paired-pulse transcranial magnetic stimulation (TMS) paradigm producing stimulus-response (S-R) curves of intracortical excitability. S-R curves were then compared with other commonly used TMS measures of cortical excitability [cortical silent period (CSP) and motor threshold (MT)]. In patients with IGE, inhibition occurred only at higher conditioning SIs compared to patients with focal epilepsy and controls. Recruitment of inhibition was unchanged in patients with focal epilepsy compared to controls. Recruitment of facilitation (ICF), CSP duration and MT, were not different between patients with FE or IGE or between patients and controls. These results suggest that the recruitment for motor cortical inhibition in patients with IGE is less effective. This may reflect a disturbed access to or an increased threshold of inhibitory neurons within the motor cortex. Impaired recruitment of inhibition might be a helpful parameter to access cortical excitability in newly diagnosed patients with generalized or focal epilepsy. PMID- 19286352 TI - Dendritic cells as controllers of antigen-specific Foxp3+ regulatory T cells. AB - Regulatory T cells (Treg) are a subpopulation of CD4(+) lymphocytes that maintain immunological self-tolerance in the periphery. Treg also regulate or suppress other classes of immune response such as allograft rejection, allergy, tumor immunity, and responses to microbes. Treg express the Foxp3 transcription factor and CD25, the high affinity interleukin-2 receptor (IL-2R). Treg are divided into two types: naturally occurring Treg derived from thymus (natural Treg) and Treg induced from Foxp3(-) CD4(+) T cells in the periphery (induced Treg). It would be valuable to understand how to control the generation of antigen-specific Treg, which could also provide a new approach to treat autoimmunity, allergy or allograft rejection without suppressing immune responses to tumor and microbes. In this review, we will discuss the role of dendritic cells (DCs) in controlling antigen-specific natural Treg and induced Treg. Natural Treg are anergic upon T cell receptor stimulation generally, however, we found that the antigen-specific natural Treg can be expanded by antigen-presenting mature bone marrow-derived dendritic cells (BM-DCs). Furthermore, recent studies showed that antigen specific Treg can be induced from Foxp3(-) CD25(-) CD4(+) T cells by antigen presenting DCs, particularly select subsets of DCs in the periphery. These findings need to be pursued to develop novel immune suppressive therapies using antigen-specific Treg educated by DCs. PMID- 19286353 TI - CYP46A1 variants influence Alzheimer's disease risk and brain cholesterol metabolism. AB - BACKGROUND: Cholesterol 24S-hydroxylase (CYP46) catalyzes the conversion of cholesterol to 24S-hydroxycholesterol, the primary cerebral cholesterol elimination product. Only few gene variations in CYP46 gene (CYP46A1) have been investigated for their relevance as genetic risk factors of Alzheimer's disease (AD) and results are contradictory. METHODS: We performed a gene variability screening in CYP46A1 and investigated the effect of gene variants on the risk of AD and on CSF levels of cholesterol and 24S-hydroxycholesterol. RESULTS: Two of the identified 16 SNPs in CYP46A1 influenced AD risk in our study (rs7157609: p=0.016; rs4900442: p=0.019). The interaction term of both SNPs was also associated with an increased risk of AD (p=0.006). Haplotypes including both SNPs were calculated and haplotype G-C was identified to influence the risk of AD (p=0.005). AD patients and non-demented controls, who were carriers of the G-C haplotype, presented with reduced CSF levels of 24S-hydroxycholesterol (p=0.001) and cholesterol (p<0.001). CONCLUSION: Our results suggest that CYP46A1 gene variations might act as risk factor for AD via an influence on brain cholesterol metabolism. PMID- 19286354 TI - Identifying predictors for good lithium response - a retrospective analysis of 100 patients with bipolar disorder using a life-charting method. AB - PURPOSE: Our aim was to investigate bipolar patients in order to test the validity of various outcome measures and to identify prognostic predictors for pharmacological treatment. MATERIAL AND METHOD: One hundred patients were interviewed using a computerized life-charting program in a descriptive, retrospective analysis. The concept "Burden of illness" was defined as a combination of severity and duration of episodes. Response to treatment was defined as the difference in burden before and after treatment, a low burden during treatment, and freedom of episodes for at least 3 years after insertion of treatment. RESULTS: The absence of mixed episodes and a high initial burden predicted a good response measured as the difference in burden. If remission for 3 years or a low burden during lithium treatment was used, the absence of rapid cycling and of mixed episodes were the most important predictors. The severity of illness before treatment had no impact. DISCUSSION AND CONCLUSION: We suggest the use of absolute measures of severity during treatment as the most appropriate measure of the outcome. Furthermore, our data provide corroboration that treatment with lithium ameliorates the prognosis of the illness, but that mixed episodes and rapid cycling predict a poorer response to lithium. PMID- 19286355 TI - Investigation on the Beckmann rearrangement reaction catalyzed by porous solids: MAS NMR and theoretical calculations. AB - In the last years, 'in situ' solid-state NMR has been applied to investigate the Beckmann rearrangement of oximes into amides using zeolites and mesoporous materials of different structure containing Bronsted acids or silanol groups as active sites. DFT methods have been applied to model the geometry of the complexes resulting from adsorption of reactants, reaction intermediates and products on clusters representing the zeolite centers, and their (15)N and (13)C NMR chemical shift calculated theoretically. This article reviews the results reported in the bibliography on the Beckmann rearrangement of various oximes (acetophenone oxime, cyclohexanone oxime and cyclododecanone oxime) mainly using 'in situ'(15)N NMR spectroscopy and theoretical calculations, and are compared with those obtained by 'in situ' infrared spectroscopy. The combination of experiment and theory has been shown to be very useful for the interpretation of the NMR spectra and the identification of the species present at different reaction temperatures. PMID- 19286356 TI - (1)H and (13)C MAS NMR studies of light alkanes activation over MFI zeolite modified by Zn vapour. AB - The early stages of methane, ethane and propane conversion were studied by in situ (1)H and (13)C MAS NMR techniques over fully exchanged Zn(2+)/MFI catalyst obtained by the reaction with zinc vapour. The in situ techniques revealed strong interaction of alkanes with Zn(2+) cations evidenced by significant shift of the corresponding NMR lines. Besides that, the formation of methyl zinc, ethyl zinc and n-propyl zinc species along with bridging and silanol surface OH-groups was detected already at the ambient temperature. These results pointed to dissociative adsorption of alkanes over (ZO)-Zn(2+)-(OZ) and (ZO)-Zn(2+)-(OSi) active sites of the catalyst. The dissociative adsorption was shown to be a dead end surface reaction in the case of methane starting reactant, while in the case of ethane and propane, it appeared to be responsible for the initiation of the catalytic cycle leading to alkenes and dihydrogen formation and regeneration of zinc containing catalytic sites. PMID- 19286357 TI - Implications of regular solution theory on the release mechanism of catanionic mixtures from gels. AB - The aim of this study was to apply the regular solution theory of mixed micelles to gain new insights on the drug release mechanism, when using catanionic mixtures as a method of obtaining prolonged release from gels. Synergistic effects were investigated at equilibrium and quantified in terms of regular solution theory interaction parameters. The drug release from catanionic aggregates was studied both in a polymer free environment, using dialysis membranes, and in gels, using a modified USP paddle method. The drug release kinetics was modelled theoretically by combining the regular solution theory with Fick's diffusion laws assuming a contribution to the transport only from monomeric species (stationary aggregates). The theoretical predictions were found to be in reasonably good agreement with experiments. An analysis of the calculated distribution of species between aggregated and monomeric states was shown to provide further insights into the release mechanism. PMID- 19286358 TI - Immuno-laser capture microdissection of frozen prolactioma sections to prepare proteomic samples. AB - Laser capture microdissection (LCM) technology combined with immunohistochemistry (immuno-LCM) is a valuable tool to obtain specific target cell populations and therefore this technique enables more accurate proteomic profile. In this study, we optimized the regular immuno-LCM technique to isolate and stain pure prolactin cells from either normal human pituitary (n=6) or prolactioma (n=11). Compared with the routine procedure, more intense and specific staining could be obtained when sections were pretreated with 0.2% Triton X-100 for 4 min. Interestingly, longer pretreatment (0.2% Triton X-100 for 10 min) or higher concentration (2% Triton X-100 for 4 and 10 min) greatly impaired labeling intensity and cell shape. Further scanning electron microscope study revealed that the component extracted from the cell surface by Triton X-100 was lipid. Using the optimized immuno-LCM technique, more pure prolactin cells could be isolated and prepared for further proteomic analysis. Taken together, we reported an optimized immuno LCM technique that could effectively dissect pure target cells in different type pituitary adenomas for further proteomics analysis. PMID- 19286359 TI - Stem cells based transplantation for cardiovascular diseases in China. AB - Stem cells based therapy has been a realistic option for cardiovascular diseases. Since 1990s, Chinese researchers and doctors have been starting to seek for optimal stem cells sources, effective methods of stem cells proliferation and differentiation with traditional Chinese medicine and clinical application of stem cells based transplantation for cardiovascular diseases. This review will summarize the investigation of stem cells in the field of cardiovascular diseases in China. PMID- 19286360 TI - Effects of late postconditioning on gene expression and cell death in neonatal rat cardiomyocyte cultures. AB - The cell death and gene expression in neonatal cardiomyocyte cultures were investigated in a late postconditioning model. The primary cultures were subjected to a 30min of anoxia followed by 60min or 24h of reoxygenation. Postconditioning was carried out in three cycles of 1min reoxygenations followed by 1min anoxia, respectively. After 24h of reperfusion the percentages of living, necrotic, and apoptotic cells were determined by staining with bis-benzimide and propidium iodide. Anoxia-reoxygenation significantly increased the necrotic and apoptotic cells both at its first and second episodes. Postconditioning in remote period did not protect the cells from the second anoxia. Postconditioning decreased the anoxia-reoxygenation-induced increase of HSP70 and HSP90 mRNA expression. We observed a decrease of HIF-3alpha gene expression in remote postconditioning. The FRAP gene expression was leveled to control value. Thus, the changes of mRNA gene expression did not show cytoprotection of cardiomyocytes in remote postconditioning model. PMID- 19286361 TI - Safety, pharmacodynamics and pharmacokinetics of TPI 1020 in smokers with asthma. AB - BACKGROUND: TPI 1020 is a novel compound with potential for anti-neutrophil effects. TPI 1020 exerts its effects by a dual mechanism of action involving corticosteroid activity and controlled donation of nitric oxide. OBJECTIVES: We assessed the safety, pharmacodynamic and pharmacokinetic activity of ascending doses of TPI 1020 compared to budesonide in asthma. METHODS: Smokers with mild asthma (n=27) were randomized to receive either 600mcg of TPI 1020 (n=13) or 400mcg of budesonide (n=14) bid for 2weeks followed by 1200 and 800mcg bid, respectively, for an additional week. RESULT: There was no serious adverse event and all but one adverse event were mild or moderate (severe headache with budesonide). Patients receiving TPI 1020 reported three-fold fewer treatment emergent AEs (n=13) than those receiving budesonide (n=39). TPI 1020 had similar effects as budesonide on FEV(1), PEF, rescue medication, asthma scoring system, methacholine response, sputum eosinophils and exhaled NO. Sputum neutrophils (%) tended to decrease more with TPI 1020 (32.6% decrease versus 3.7% increase for budesonide); the decrease occurring only in patients with high neutrophils at baseline. A significant difference favoring TPI 1020 was noted for CRP. Budesonide caused a statistically significant decrease in 24h urinary free cortisol over 22days (median of 4.4-2.8mcg/ml, p=0.01) whereas TPI 1020 had no such effect (4.4-5.8mcg/ml), suggesting lower systemic corticosteroid exposure following TPI 1020 treatment. CONCLUSION: TPI 1020 appears safe in asthmatic smokers and warrants further investigation in respiratory conditions. PMID- 19286362 TI - Real-life experience of older patients requiring non-invasive ventilation for exacerbations of chronic obstructive pulmonary disease. PMID- 19286363 TI - The role of RNP biogenesis in spinal muscular atrophy. AB - Mutations that affect pre-mRNA processing are the cause for many genetic diseases. Most such mutations target cis-acting regulatory sequences in a given transcript, thus preventing its proper maturation. Only recently however, mutations in trans-acting factors involved in pre-mRNA processing have likewise been linked to disease. One prominent example is spinal muscular atrophy (SMA), a monogenic, neuromuscular disorder caused by reduced levels of functional survival motor neuron (SMN) protein. This ubiquitous factor is part of a complex that mediates the formation of spliceosomal snRNPs. The detailed biochemical investigation of SMN under normal conditions and in SMA has provided clues how mutations in factors with general functions elicit tissue-specific phenotypes. PMID- 19286364 TI - Non-invasive glucose monitoring in patients with Type 1 diabetes: a Multisensor system combining sensors for dielectric and optical characterisation of skin. AB - In vivo variations of blood glucose (BG) are affecting the biophysical characteristics (e.g. dielectric and optical) of skin and underlying tissue (SAUT) at various frequencies. However, the skin impedance spectra for instance can also be affected by other factors, perturbing the glucose related information, factors such as temperature, skin moisture and sweat, blood perfusion as well as body movements affecting the sensor-skin contact. In order to be able to correct for such perturbing factors, a Multisensor system was developed including sensors to measure the identified factors. To evaluate the quality of glucose monitoring, the Multisensor was applied in 10 patients with Type 1 diabetes. Glucose was administered orally to induce hyperglycaemic excursions at two different study visits. For analysis of the sensor signals, a global multiple linear regression model was derived. The respective coefficients of the variables were determined from the sensor signals of this first study visit (R(2)=0.74, MARD=18.0%--mean absolute relative difference). The identical set of modelling coefficients of the first study visit was re-applied to the test data of the second study visit to evaluate the predictive power of the model (R(2)=0.68, MARD=27.3%). It appears as if the Multisensor together with the global linear regression model applied, allows for tracking glucose changes non invasively in patients with diabetes without requiring new model coefficients for each visit. Confirmation of these findings in a larger study group and under less experimentally controlled conditions is required for understanding whether a global parameterisation routine is feasible. PMID- 19286365 TI - A systematic investigation into the electrical properties of single HeLa cells via impedance measurements and COMSOL simulations. AB - The electrical properties of single cells provide fundamental insights into their pathological condition and are therefore of immense interest to medical practitioners. Accordingly, this study captures single HeLa cells using a microfluidic device and then measures their impedance properties using a commercial impedance spectroscopy system. The experimental system is modeled by an equivalent electrical circuit and COMSOL simulations are then performed to establish the conductivity, permittivity and impedance of single HeLa cells under various operational frequencies and voltages. At an operational voltage of 0.2 V, the maximum deviation between the experimental and simulation results for the magnitude and phase of the HeLa cell impedance is found to be 9.5% and 4.2%, respectively. In general, both sets of results show that the conductivity and permittivity of single HeLa cells increase with an increasing operational voltage. Moreover, an increasing frequency is found to increase the conductivity of HeLa cells at all values of the operational voltage, but to reduce the permittivity for operational voltages in the range 0.6-1.0 V. Based upon the simulation and experimental results, empirical equations are constructed to predict the conductivity and permittivity of single HeLa cells under specified values of the operational voltage and frequency, respectively. The maximum discrepancy between the predicted results and the simulation results for the permittivity and conductivity of the HeLa cells at an operational voltage of 0.2 V is found to be just 0.5% and 4.5%, respectively. PMID- 19286366 TI - Structural biology of the p53 tumour suppressor. AB - The p53 tumour suppressor protein has presented a challenge for structural biology for more than two decades. The complete p53 molecule has eluded numerous attempts to determine its structure, presumably owing to the intrinsic conformational flexibility that is essential to the protein's function. Recent data obtained by X-ray crystallography, NMR spectroscopy and electron microscopy provide new insight into the quaternary architecture of the whole molecule and new strategies for examining how these structures correlate with the cell and molecular biology of the 'Guardian of the Genome'. PMID- 19286367 TI - Toward an atomic model of the 26S proteasome. AB - Since the discovery of the 26S proteasome, much progress has been made in determining the structure of this large dynamic protein complex. Until now, a vast amount of structural information of the proteasome has been obtained from all kinds of structure determination techniques, and the function of the protease core is well understood at atomic detail. Yet our understanding of the entire 26S proteasome structure, particularly its 19S regulatory complex, is still limited at a low-resolution blob-ology level. In this review, we highlight the recent progress made in understanding the mechanism of 20S gate opening by the proteasomal activators. We also emphasized the recent methodological advances, particularly in achieving the near atomic resolution by single particle electron cryomicroscopy, and the possible approaches that will enable more detailed structural analysis of the entire 26S proteasome. PMID- 19286368 TI - Adjuvant treatment of GIST with imatinib: solid ground or still quicksand? A comment on behalf of the EORTC Soft Tissue and Bone Sarcoma Group, the Italian Sarcoma Group, the NCRI Sarcoma Clinical Studies Group (UK), the Japanese Study Group on GIST, the French Sarcoma Group and the Spanish Sarcoma Group (GEIS). PMID- 19286369 TI - The prognostic significance of inflammation and medullary histological type in invasive carcinoma of the breast. AB - The new gene expression molecular taxonomy of breast cancer places medullary carcinoma in the basal group. The basal group is considered to have a poor prognosis, but medullary carcinoma is considered to have a better prognosis than other grade 3 carcinomas. The prognostic significance of tumour associated inflammation, an important feature of medullary carcinomas, remains controversial. The aim of this study was to assess the prognostic importance of medullary histological type and inflammation in breast cancer. One thousand five hundred and ninety-seven patients who received no systemic adjuvant treatment and who had a median follow up of 9.5 years were studied. RESULTS: Prominent inflammation was associated with high histological grade and with better survival [relative risk (RR) 0.57, 95% confidence intervals (CI) 0.44-0.74] on multivariate analysis. Typical and atypical medullary carcinomas (n=132) did not have significantly different survival and were grouped together. Medullary carcinoma did not have significantly different prognosis than grade 3 ductal carcinoma with prominent inflammation, but both had a better prognosis than grade 3 ductal carcinoma without prominent inflammation (P<0.0001 and P=0.03). These differences were independent of other prognostic factors. These results question the current separation of typical and atypical medullary carcinoma. Prominent inflammation is associated with a better prognosis, and may explain the better prognosis in medullary carcinoma compared with grade 3 ductal carcinoma without prominent inflammation. The good prognosis of medullary carcinoma emphasises the heterogeneity of basal-like breast carcinomas. Further studies are needed to investigate the difference in survival between medullary carcinoma and other forms of basal carcinomas and the role of inflammation in any such differences in behaviour. PMID- 19286370 TI - Open, laparoscopic and robotic radical prostatectomy: optimizing the surgical approach. AB - As advances in the understanding of prostatic anatomy led to improvements in functional and oncologic outcomes after prostatectomy of the past few decades, advances in technology and surgical technique have made minimally-invasive prostate surgery a reality. Today patients diagnosed with clinically localized prostate cancer have more surgical treatment options than in the past including open, laparoscopic and robot-assisted laparoscopic radical prostatectomy. Advantages and disadvantages exist for each modality and lead to subtle differences in the technical execution of the procedure. Evidence from centers of excellence and from experienced surgeons demonstrates that both laparoscopic and robotic-assisted laparoscopic radical prostatectomy appear to be comparable to outcomes achieved with open radical retropubic prostatectomy series. Individual surgeon skill, experience and clinical judgment are likely the stronger predictors of outcome rather than the technique chosen. However, learning curves, oncologic outcomes and cost-efficacy remain important considerations in the dissemination of minimally-invasive prostate surgery. A greater appreciation of the periprostatic anatomy and further modification of surgical technique will result in continued improvement in functional outcomes and oncological control for patients undergoing radical prostatectomy, whether by open or minimally invasive surgery. PMID- 19286371 TI - Development of environmentally superior technologies in the US and policy. AB - "Environmentally superior technology" (EST) represents a research initiative in North Carolina (NC) to develop alternatives to lagoon treatment and land application of swine manure. EST performance standards included impacts of animal waste to surface and groundwater, emission of ammonia and odor, release of disease-transmitting pathogens, and heavy metal contamination of soil and groundwater. Five technologies were shown to meet these standards: a solids separation/nitrification-denitrification/soluble phosphorus removal system; a thermophilic anaerobic digester system; a centralized composting system; a gasification system; and a fluidized bed combustion system. Economic data compiled for all EST showed annualized (10-year) costs of retrofitting existing swine farms with the technologies ranged between $90 and over $400 per 1000 lbs. steady state live-weight. Value-engineering to reduce the costs of targeted EST has been successful. Policy providing institutional incentives to incorporate EST has been enacted as a result of this study. PMID- 19286372 TI - Characterization of enzymatic hydrolyzed snow crab (Chionoecetes opilio) by product fractions: a source of high-valued biomolecules. AB - Snow crab (Chionoecetes opilio) constitutes valuable and nutritional sources of components, such as proteins, lipids and chitin. The present investigation was undertaken to evaluate the feasibility of applying a pilot scale enzymatic hydrolysis process of snow crab by-products, followed by fractionation, in order to recover enriched high-valued compounds. The yield of snow crab by-products recovered after manual processing; on a dry weight was 87.4%. The by-products (raw materials) were mainly moist (approximately 78%), and contained 42.9% proteins, 14.8% lipids, 25.7% minerals, 16.2% chitin, all expressed on a dry weight. The fatty acid profile of snow crab by-products and all fractions obtained following processing showed a higher content in mono-unsaturated fatty acids (MUFAs; approximately 50%), followed by polyunsaturated fatty acids (PUFAs; approximately 20%) and saturated fatty acids (SFAs; approximately 15%). The n-3/n 6 ratio was approximately 10 and represents a good index of nutritional value for snow crab oil by-products. Most protein enriched fractions demonstrate a well balanced amino acid composition, notably the most essential amino acids. These protein fractions are characterized by biomolecules having a relatively low molecular weight (35 kDa and less) range. The enzymatic hydrolysis process developed in this study shows that snow crab by-products should to be viewed as having the potential of being identified as high-valued products. Even though the process could be optimized, it is controllable, and depending on hydrolyses conditions, the products obtained are reproducible and well defined. Results presented in this study indicate that snow crab by-products may serve as excellent nutritional components for future applications in the health and food sectors. PMID- 19286373 TI - Development of a second-generation environmentally superior technology for treatment of swine manure in the USA. AB - New swine waste management systems in North Carolina need to meet high performance standards of an environmentally superior technology (EST) regarding nitrogen, phosphorus, heavy metals, pathogens, ammonia and odor emissions, and remain affordable and simple to operate. The objective of this study was to develop a second-generation treatment system that can achieve high EST standards at reduced costs. The system used solids separation, nitrification/denitrification and phosphorus removal/disinfection, and was demonstrated at full-scale on a 5145-head swine farm during three production cycles (15-months). Removal efficiencies were: 98% suspended solids, 97% ammonia, 95% phosphorus, 99% copper and zinc, 99.9% odors, and 99.99% pathogens. The system met EST standards at 1/3 the cost of the previous version. Animal health and productivity were enhanced; hog sales increased 32,900 kg/cycle (5.6%). These results demonstrated that: (1) significant cost reductions were achieved by on farm implementation and continued engineering improvements, and (2) the new waste management system substantially benefited livestock productivity. PMID- 19286374 TI - Predicting ultimate methane yields of Jatropha curcus and Morus indica from their chemical composition. AB - In this study, all the components of Jatropha curcus and Morus indica were chemically characterized and their biochemical methane potentials (BMP) were determined. From the variables that showed strong influence on the ultimate methane yield (B(o)) of J. curcus, a multiple regression Jatropha model was developed. This model comprised of total carbohydrates, protein, lipid, acid detergent fiber (ADF), cellulose and ash in ADF as independent variables, with r(2) value of 0.943. The Jatropha model was validated on 7 samples of M. indica parts and wastes from silkworm rearing trays of this study and 13 samples of heterogeneous organic wastes of earlier studies, to judge the prediction quality. It was found that most of the predicted values differed by less than 15% of their experimental B(o). PMID- 19286375 TI - Synthesis and cytotoxicity of 2-cyano-28-hydroxy-lup-1-en-3-ones. AB - New A-ring modified betulin and dihydrobetulin derivatives possessing the 2-cyano 1-en-3-one moiety were prepared and tested for cytotoxicity in seven cancer cell lines. The most active agent 9a synthesized in this account was further demonstrated to induce apoptosis and to activate caspases in malignant melanoma cells. PMID- 19286376 TI - Fluorinated non-imidazole histamine H3 receptor antagonists. AB - Fluorine substituents have become a widespread and important component in drug design and development. Here, the synthesis of fluorine containing compounds and some corresponding precursor molecules are presented for potential isotope labelling as well as their data obtained with in vitro and in vivo screenings. The compounds vary in the basic centres (piperidine or pyrrolidine) and are fluoro substituted in different positions of the basic alicyclic moiety. Pharmacological evaluation resulted in ligands with high affinities at hH(3) receptor in the nanomolar and subnanomolar concentration range and some with high antagonist in vivo potencies. PMID- 19286377 TI - 8-[2-(4-Aryl-1-piperazinyl)ethyl]-2H-1,4-benzoxazin-3(4H)-ones: dual-acting 5-HT1 receptor antagonists and serotonin reuptake inhibitors--part II. AB - 8-[2-(4-Aryl-1-piperazinyl)ethyl]-2H-1,4-benzoxazin-3(4H)-ones have been identified as highly potent 5-HT(1A/B/D) receptor antagonists with and without additional SerT activity and a high degree of selectivity over hERG potassium channels. Modulation of the different target activities gave compounds with a range of profiles suitable for further in vivo characterization. PMID- 19286378 TI - Inhibition of carbonic anhydrase isozymes with benzene sulfonamides incorporating thio, sulfinyl and sulfonyl glycoside moieties. AB - A series of benzene sulfonamides incorporating thio, sulfinyl or sulfonyl glycoside moieties were synthesized. These glycoconjugates were investigated for their ability to inhibit the enzymatic activity of four human carbonic anhydrases (hCA): isozymes I, II and tumour-associated isozymes IX and XII. The oxidation state of the sulfur in the carbohydrate tail moiety did not influence either enzyme inhibition potency or isozyme selectivity even though presenting opportunities for differing interactions with the target isozymes. PMID- 19286379 TI - Synthesis and anti-inflammatory activity of the major metabolites of imrecoxib. AB - We have developed a novel and moderately selective COX-2 inhibitor, imrecoxib, as a new anti-inflammatory drug. We describe herein the preparation of the major metabolites M2 and M4 of imrecoxib, as well as the in vitro and in vivo activities of the two compounds. The results showed that both M2 and M4 are potential COXs inhibitors with a moderate COX-1/COX-2 selectivity, and their anti inflammatory activity in vivo was equal to or slightly higher than the clinical celecoxib. PMID- 19286380 TI - Diphenyl ethers as androgen receptor antagonists for the topical suppression of sebum production. AB - A series of diphenyl ethers was prepared and evaluated for androgen receptor antagonist activity in human androgen receptor binding and cellular functional assays. Analogs with potent in vitro activities were evaluated for topical in vivo efficacy in the Golden Syrian Hamster ear model. Several compounds showed reduction in wax esters in this validated animal model. PMID- 19286381 TI - Synthesis and evaluation of 2,7-diamino-thiazolo[4,5-d] pyrimidine analogues as anti-tumor epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors. AB - 2,7-Diamino-thiazolo[4,5-d]pyrimidine analogues were synthesized as novel epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors. Representative compounds showed potent and selective EGFR inhibitory activities and inhibited in vitro cellular proliferation in EGFR-overexpressing human tumor cells. The synthesis and preliminary biological, physical, and pharmacokinetic evaluation of these thiazolopyrimidine compounds are reported. PMID- 19286382 TI - Olfactory stimuli and enhanced postural stability in older adults. AB - Ameliorating postural instability is an important component of geriatric health care. The effect of olfactory stimuli (lavender and black pepper oils) on postural control in 17 older adults (78+/-6 years old) who had no apparent neurological deficits was studied. Measurements of center of pressure (CoP) trajectories were done with subjects standing quietly on a force plate. Control measurements were compared with olfactory interventions: brief exposure to sham (distilled water), lavender oil, and black pepper oil; experiments were repeated with eyes open and eyes closed. From the CoP data, the root mean square (RMS) displacement and velocity in mediolateral (ML) and antero-posterior (AP) directions, and the total trajectory length were computed. This study found that with eyes closed, olfactory stimulation with either lavender or black pepper oil significantly decreased both ML and AP RMS velocities and trajectory lengths compared with baseline. In contrast, little effect was observed under the eyes open condition. Decreases in RMS displacements were small and mostly insignificant. The study suggests that olfactory stimulation may improve posture stability in older adults through decreasing the velocities of postural adjustments during normal sway. PMID- 19286383 TI - Skeletal fluorosis causing high cervical myelopathy. AB - Skeletal fluorosis is endemic in some parts of the world and is the result of life-long ingestion of high amounts of fluoride in drinking water. Its clinical presentation is characterized mostly by bone and dental changes with later ossification of many ligaments and interosseous membranes. We present a rare case of high cervical myelopathy caused by ossification of the posterior longitudinal ligament and ligamentum flavum in a patient from an area endemic for skeletal fluorosis. The clinical presentation of skeletal fluorosis and treatment options are discussed. PMID- 19286384 TI - The neurofilament light chain gene (NEFL) mutation Pro22Ser can be associated with mixed axonal and demyelinating neuropathy. AB - We report the detailed clinical, electrophysiological and molecular analysis of a patient with Charcot-Marie-Tooth (CMT) disease. DNA sequencing of the coding sequences of the neurofilament light chain polypeptide (NEFL) gene revealed a c.64C>T heterozygous, missense mutation resulting in a Pro22Ser amino acid substitution. Clinical and electrophysiological studies revealed a mixed axonal and demyelinating neuropathy, with widespread demyelination involving both proximal and distal nerve segments. Mutations at this site in the NEFL gene have been previously linked to an axonal neuropathy or distal nerve demyelination. Our results emphasize the complexity of genotype-phenotype correlations in CMT and underline the possible importance of host factors and gene interactions in the development of clinical phenotypes. PMID- 19286385 TI - Overnight switching from ergot-derived dopamine agonists to pramipexole in patients with Parkinson's disease: an open preliminary trial in Japan. AB - The aim of this study was to evaluate the safety and tolerability of overnight switching from ergot-derived dopamine agonists such as cabergoline to equivalent doses of pramipexole in patients with Parkinson's disease. The safety of overnight switching to pramipexole from cabergoline, which has a long plasma half life and may cause dopaminergic excess after switchover, has not been established. Twenty-two consecutive patients with Parkinson's disease were included, 18 of them on cabergoline treatment. Patients were assessed with the Unified Parkinson's Disease Rating Scale (UPDRS) given just prior to switching as well as after 2, 4, 8, and 12 weeks of treatment. Eight patients (36.4%) experienced adverse events, of whom two were withdrawn from the study. Generally, however, significant improvement in the UPDRS was obtained after 2 weeks and improvement was maintained up to 12 weeks of treatment. Therefore, our study showed that overnight switching from ergot-derived dopamine agonists including cabergoline to dose-equivalent pramipexole was safe when associated with good patient compliance. PMID- 19286386 TI - Incorporation of an inducible nucleotide analog into DNA by DNA polymerases. AB - Non-natural nucleotides with diverse functionalities are highly useful in many areas of basic research and practical applications. We have previously developed an efficient method for post-synthetic modifications of 2-amino-6-vinylpurine (AVP)-containing oligonucleotides, which permits conjugations of a variety of useful functional appendages to the AVP moiety in DNA. Here we report an investigation on the ability of various DNA polymerases to use 5'-triphosphate of 2'-deoxyribosyl-2-amino-6-(2-methylthioethyl)purine (a stable precursor of AVP) as the substrate for templated DNA synthesis. PMID- 19286387 TI - Nanoindentation of gold nanoparticles functionalized with proteins. AB - The hardness and Young's modulus of 10 and 20 nm gold nanoparticles (Au NPs) modified with bovine serum albumin and streptavidin were measured using a nanoindenter. The Au NPs were immobilized on a semiconductor surface through organic self-assembled monolayers. Changes in mechanical properties occurred when the Au NPs were immobilized on the surface. The hardness and Young's modulus were dependent on the size of the NPs, and the proteins on the particles showed highly plastic and elastic behavior compared to flat surfaces modified with self assembled monolayers. PMID- 19286388 TI - Quality of life evolution after lung cancer surgery in septuagenarians: a prospective study. AB - OBJECTIVE: To prospectively evaluate quality of life (QoL) evolution after lung cancer surgery in a cohort of septuagenarians with the European Organisation for Research and Treatment of Cancer (EORTC) QoL Questionnaire-C30 and LC13. METHODS: Between January 2003 and December 2006, QoL was prospectively recorded in 60 consecutive septuagenarians undergoing lung cancer surgery. Forty-nine lobectomies and 11 pneumonectomies were performed. Questionnaires were administered before surgery and 1, 3, 6 and 12 months postoperatively (MPO) with response rates of 100%, 83%, 87%, 90% and 77%, respectively. RESULTS: After lobectomy, QoL scores returned to baseline 3-6 months after surgery, with the exception of a persistent decrease in physical functioning and an increase in dyspnea within the 12 months follow-up. In the 12 months follow-up period after pneumonectomy, there was no return to baseline in physical, role and social functioning. After pneumonectomy, most quality of life scores returned to baseline at 1-month follow-up, with the exception of dyspnea and general pain, which returned to baseline at 3 and 6 months, respectively. Comparing both resections, significant differences in evolution of physical functioning (6MPO p=0.045), role functioning (3MPO p=0.035), social functioning (6MPO p=0.006, 12MPO p=0.001) and general pain (6MPO p=0.037) were reported in favor of lobectomy. CONCLUSIONS: The present study documented QoL evolution profiles of septuagenarians after pulmonary surgery. The results indicate that both resections have a major impact on elderly patients, especially physical functioning and dyspnea status. If both resections are compared, lobectomy patients have a more favorable evolution in QoL subscales compared to pneumonectomy. PMID- 19286389 TI - Tumors that look for their springtime in APRIL. AB - Inflammatory cells produce a proliferation inducing ligand (APRIL), one of the most recently cloned members of the tumor necrosis factor (TNF) family. Early experiments implicated APRIL as a promoting factor in the natural course of various cancers, reinforcing the concept that host inflammatory reactions are part of a tumor development. Recent studies have further analyzed the tumor promoting role of APRIL in patients with solid tumors or with hematological malignancies. Here, we will review the recent literature, and provide evidence that APRIL may be a useful prognostic tool and a potential target in the treatment of some cancers. PMID- 19286390 TI - Synergistic effects of granulocyte-colony stimulating factor on bone marrow stromal cell transplantation for mice cerebral infarct. AB - This study was aimed to assess whether ex vivo treatment with granulocyte-colony stimulating factor (G-CSF) modifies biological properties of bone marrow stromal cells (BMSC) and enhances functional recovery by BMSC transplantation into infarct brain. Immunohistochemistry was conducted to characterize the cultured BMSC. The pharmacological effects of G-CSF on their proliferation, cell cycle, and growth factor production were precisely analyzed, using FACS and ELISA techniques. Non-treated or G-CSF treated BMSC were stereotactically transplanted into the mice brain subjected to cerebral infarct, and its effects on functional and histological aspects were evaluated. The BMSC expressed the receptor for G CSF. Treatment with 0.1muM of G-CSF significantly enhanced the proliferation of BMSC by increasing their population in S phase, and increased their production of SDF-1alpha, HGF, and NGF. When transplanted into infarct brain, G-CSF treated BMSC significantly improved motor function as early as 2 weeks after transplantation, whereas non-treated BMSC did 4 weeks after transplantation. These findings strongly suggest that G-CSF may enhance the proliferation and growth factor production of the cultured BMSC and accelerate functional restoration by BMSC transplantation. Such pharmacological "activation" of the BMSC may contribute to successful clinical application of BMSC transplantation therapy for ischemic stroke. PMID- 19286391 TI - Echinacea tennesseensis ethanol tinctures harbor cytokine- and proliferation enhancing capacities. AB - BACKGROUND: Members of the genus Echinacea are used medicinally to treat upper respiratory infections such as colds and influenza. The aim of the present investigation was to characterize the phytomedicinal properties of the American federally endangered species Echinacea tennesseensis. METHODS: Fifty-percent ethanol tinctures were prepared from roots, stems, leaves, and flowers and tested separately for their ability to influence production of IL-1beta, IL-2, IL-10, and TNF-alpha as well as proliferation by young human adult peripheral blood mononuclear cells (PMBC) in vitro. Tincture aliquots were stored at three different temperatures (4, -20, and -80 degrees C) for 21h before testing. At 1 month post-extraction, tinctures stored at -20 degrees C were tested again for cytokine modulation. Phytochemical analyses were performed using HPLC. RESULTS: Fresh root, leaf, and flower tinctures stimulated PBMC proliferation. Fresh root tinctures alone stimulated IL-1beta, IL-10, and TNF-alpha production. No tinctures modulated IL-2 production. Stem tinctures showed no activity. Storage temperature did not influence any outcomes. Root tinctures maintained their ability to modulate IL-1beta, IL-10, and TNF-alpha production after 1month of storage at -20 degrees C. CONCLUSIONS: These results suggest E. tennesseensis harbors phytomedicinal properties that vary by plant organ, with roots demonstrating the strongest activities. PMID- 19286392 TI - TNF-alpha gene expression in colorectal mucosa as a predictor of remission after induction therapy with infliximab in ulcerative colitis. AB - BACKGROUND: It has been documented that treatment with infliximab (IFX) induces remission in 1/3 of patients with moderate to severe ulcerative colitis (UC). Predictors of response could improve selection of patients with a higher probability of favorable outcome. AIM: To determine predictor factors for the clinical outcome of IFX induction therapy in UC. METHODS: UC patients with moderate to severe disease who received 5mg/kg IFX at weeks 0, 2 and 6weeks were included. Ulcerative colitis disease activity index (UCDAI) score including endoscopic sub-scores were assessed before and after treatment. Several predictors, including TNF-alpha mRNA expression, were tested in a regression model. RESULTS: Fifty-nine patients completed the study. Age, gender, steroid therapy, immunosuppressive, pancolitis, endoscopic sub-score, disease duration, C reactive protein, interleukin-(IL)-4, IL-10 or interferon-gamma (IFN-gamma) did not predict mucosal or clinical remission. There was an inverse and independent association between pre-treatment TNF-alpha expression levels and clinical and endoscopic remission of IFX treatment (logistic regression, p=0.01 and p=0.003, odds ratio 2.5 and 4.8, respectively). CONCLUSION: The clinical outcome of an induction therapy with IFX in UC is inversely associated with the pre-treatment gene expression levels of TNF-alpha in colorectal mucosa. PMID- 19286393 TI - Direct determination of the primary binding site of cisplatin on cytochrome C by mass spectrometry. AB - Protein-cisplatin interactions lie at the heart of both the effectiveness of cisplatin as a therapeutic agent and side effects associated with cisplatin treatment. Because a greater understanding of the protein-cisplatin interactions at the molecular level can inform the design of cisplatin-like agents for future use, mass spectrometric determination of the binding site of cisplatin on a model protein, cytochrome c, was undertaken in this paper. The monoadduct cytochrome c Pt(NH(3))(2)(H(2)O) is found to be the primary adduct produced by the cytochrome c-cisplatin interactions under native conditions. To locate the primary binding site of cisplatin, both free cytochrome c and the cytochrome c adducts underwent trypsin digestion, followed by Fourier transform mass spectrometry (FT-MS) to identify unique fragments in the adduct digest. Four such fragments were found in the adduct digest. Tandem mass spectrometry (MS/MS and MS(3) indicates that two fragments are Pt(NH(3))(2)(H(2)O) bound peptides (Gly56-Glu104 and Asn54-Glu104) with one water associated at the peptide bond Lys79-Met80, and the other two fragments are heme containing peptides (acety1-Gly1-Lys53 and acety1-Gly1-Lys55). The product-ion spectra of the four fragments reveal that Met65 is the primary binding site of cisplatin on cytochrome c. PMID- 19286394 TI - Tracking radical migration in large hydrogen deficient peptides with covalent labels: facile movement does not equal indiscriminate fragmentation. AB - Photodissociation of iodo-tyrosine modified peptides yields localized radicals on the tyrosine side chain, which can be further dissociated by collisional activation. We have performed extensive experiments on model peptides, RGYALG, RGYG, and their derivatives, to elucidate the mechanisms underlying backbone fragmentation at tyrosine. Neither acetylation nor deuteration of the tyrosyl phenolic hydrogen significantly affects backbone fragmentation. However, deuterium migration from the tyrosyl beta carbon is concomitant with cleavage at tyrosine. Substitution of tyrosine with 4-hydroxyphenylglycine, which does not have beta hydrogens, results in almost complete elimination of backbone fragmentation at tyrosine. These results suggest that a radical situated on the beta carbon is required for a-type fragmentation in hydrogen-deficient radical peptides. Replacement of the alphaH of the residue adjacent to tyrosine with methyl groups results in significant diminution of backbone fragmentation. The initial radical abstracts an alphaH from the adjacent amino acid, which is poised to "rebound" and abstract the betaH of tyrosine through a six-membered transition state. Subsequent beta-scission leads to the observed a-type backbone fragment. These results from deuterated peptides clearly reveal that radical migration in peptides can occur and that multiple migrations are not infrequent. Counterintuitively, close examination of all experimental results reveals that the probability for fragmentation at a particular residue is well correlated with thermodynamic radical stability. A-type fragmentation therefore appears to be most likely when favorable thermodynamics are combined with the relevant kinetic control. These results are consistent with ab initio calculations, which demonstrate that barriers to migration are significantly smaller in magnitude than probable dissociation thresholds. PMID- 19286395 TI - What is the double shadow in the chest? PMID- 19286396 TI - Semantic and subword priming during binocular suppression. AB - In general, stimuli that are familiar and recognizable have an advantage of predominance during binocular rivalry. Recent research has demonstrated that familiar and recognizable stimuli such as upright faces and words in a native language could break interocular suppression faster than their matched controls. In this study, a visible word prime was presented binocularly then replaced by a high-contrast dynamic noise pattern presented to one eye and either a semantically related or unrelated word was introduced to the other eye. We measured how long it took for target words to break from suppression. To investigate word-parts priming, a second experiment also included word pairs that had overlapping subword fragments. Results from both experiments consistently show that semantically related words and words that shared subword fragments were faster to gain dominance compared to unrelated words, suggesting that words, even when interocularly suppressed and invisible, can benefit from semantic and subword priming. PMID- 19286397 TI - Conservative or surgical treatment for subacromial impingement syndrome? A systematic review. AB - BACKGROUND: Patients with subacromial impingement syndrome are often operated on when conservative treatments fail. But does surgery really lead to better results than nonoperative measures? This systematic review compared effects of conservative and surgical treatment for subacromial impingement syndrome in terms of improvement of shoulder function and reduction of pain. METHODS: A literature search for randomized controlled trials (RCTs) in PubMed, EMBASE, PEDro, and the Cochrane Central Register of Controlled Trials was conducted. Two reviewers assessed the methodological quality of the selected studies. A best-evidence synthesis was used to summarize the results. RESULTS: Four RCTs were included in this review. Two RCTs had a medium methodological quality, and 2 RCTS had a low methodological quality. No differences in outcome between the treatment groups were reported for any of the studies, irrespective of quality. CONCLUSION: No high-quality RCTs are available so far to provide possible evidence for differences in outcome; therefore, no confident conclusion can be made. According to the best-evidence synthesis, however, there is no evidence from the available RCTs for differences in outcome in pain and shoulder function between conservatively and surgically treated patients with SIS. LEVEL OF EVIDENCE: Review. PMID- 19286398 TI - Cement extrusion causing radial nerve palsy after shoulder arthroplasty: a case report. PMID- 19286399 TI - Arthroscopic repair of large and massive rotator cuff tears performed under local anaesthesia and sedation. PMID- 19286400 TI - Reliability and accuracy of templating the proximal humeral component for shoulder arthroplasty. AB - BACKGROUND: The purpose of this paper is to examine the intra-observer and inter observer reliability of preoperative templating to help determine its usefulness for approximating proximal humeral stem and head size in shoulder arthroplasty. METHODS: Thirty-one nonconsecutive patients underwent uncemented total shoulder arthroplasty using 1 of 2 commonly used prostheses. Templating was performed by 4 independent observers (2 experienced shoulder surgeons and 2 PGY-3 residents). Two readings were performed with a minimum 2-week interval between templating sessions. Templating was performed in a blinded manner using appropriately magnified templates provided by the prosthesis manufacturer. Postoperative radiographs were analyzed in the same fashion and each observer graded qualitative stem and head size radiographically. Pre-operative templated sizes were compared to actual implant sizes obtained from dictated operative reports. RESULTS: There was substantial (kappa > 0.7) intraobserver reliability for stem and head size selection for both prostheses. Interobserver reliability was moderate for stem size (kappa = 0.53) and substantial (kappa = 0.66) for head size. Preoperative templating accurately predicted stem (84-95%) and head (44 66%) size within 1 size variation (stem width, head thickness, or diameter) for shoulder replacement surgery. There was no significant difference in accuracy between attendings and residents when templating for stem or head size. CONCLUSION: Preoperative templating is a reliable and reasonably accurate step when planning shoulder replacement surgery. Templating for stem size is more accurate than for head size. PMID- 19286401 TI - Characterization of porcine factor VII, X and comparison with human factor VII, X. AB - OBJECTIVE: Factor VII (FVII) and factor X (FX) are two predominant molecules of coagulation cascade. Whether porcine FVII and FX could efficiently work in human circulation is important for successful pig to human liver transplantation. We compared the genetic characterizations and coagulation activities of porcine and human FVII and FX to shed insight into the further investigation of potential inter-species molecular incompatibility between porcine FVII, FX and human derived procoagulants and anticoagulants in xenotransplantation. METHODS: Multiple rounds of PCR were used to screen the positive clones from a porcine liver tissue cDNA library. 5' RACE and 3' RACE were conducted to get the full length cDNA. The three-dimensional structure of protein was modeled by Swiss Model program. Prothrombin Time (PT) of porcine and human plasma was determined by coagulation autoanalyzer. Activities of porcine FVII and FX were detected by adding the porcine plasma into FVII or FX-deficient human plasma. RESULTS: We cloned the full-length cDNA of porcine FVII and FX, which contained 1416 bp and 1856 bp, coding 445 and 479 amino acids, respectively. Porcine FVII and FX shared 74.08% and 73.1% amino acid identities with human FVII and FX. Sequence alignments showed that porcine FVII might have additional gamma-carboxyglutamic acid in Gla domain, and one important variation of Lys62-Glu in light chain. No significant difference was observed in TF binding region of heavy chain, while 4 variations were identified in the important functional residues responsible for proteolysis activity, as Gln217-Glu, Thr151-Lys, Glu154-Val and Gln40-Leu. However, no apparent change was displayed in the 3-D model of the heavy chain of porcine FVII. When porcine FX was analyzed, great variations have been found at active peptide (Ser143 to Arg194) with only 11.6% identity. Some important variations at gamma-carboxyglutamic acids and Ca(2+) binding sites were identified, while high conservations were discovered at other functional sites. Comparisons on 3-D protein models demonstrated that the protein backbones of porcine and human FX were highly conserved, and little difference was shown at the molecular surface of anticoagulant binding sites S2 and S3. PT detection of porcine and human plasma showed similar results, while coagulation activities of porcine FVII and FX were remarkably higher than that of human. CONCLUSION: Porcine FVII and FX showed relatively high homology with human FVII and FX in nucleotide, amino acid sequences and three-dimensional structure. However, the different affinities to important macromolecules caused by genetic differences might contribute to the molecular incompatibilities in liver xenotransplantation. PMID- 19286402 TI - Up-regulated expression of peroxisome proliferator-activated receptor gamma in the hypothalamic-pituitary-adrenal axis of weaned pigs after Escherichia coli lipopolysaccharide challenge. AB - The expression of peroxisome proliferator-activated receptor gamma (PPARgamma) was investigated in the hypothalamic-pituitary-adrenal (HPA) axis of weaned pigs after injection with 100 microg/kg bodyweight Escherichia coli lipopolysaccharide (LPS) (n=6) and control pigs injected with sterile saline (n=6). LPS increased PPARgamma mRNA and protein expression in the hypothalamus (23.8 and 3.1-fold relative to controls, respectively), pituitary gland (9.2 and 2.0-fold, respectively) and adrenal gland (3.5 and 2.3-fold, respectively) (P<0.05). LPS also induced an increase in PPARgamma immunohistochemical staining in the hypothalamus (1.3-fold), adenohypophysis (1.3-fold), adrenal cortex (1.4-fold) and adrenal medulla (1.6-fold) (P<0.05). Concurrent with up-regulated expression of PPARgamma, LPS increased the concentrations of plasma corticotrophin-releasing hormone (2.1-fold) and adrenocorticotrophin (1.4-fold) (P<0.05). LPS also induced elevations of interleukin 6 and tumour necrosis factor alpha mRNA levels in the hypothalamus (4.0 and 3.2-fold, respectively), pituitary gland (20.7 and 5.1 fold, respectively) and adrenal gland (3.9 and 3.3-fold, respectively) (P<0.05). PPARgamma may play a role in the regulation of neuroendocrine responses associated with immunological stress in pigs. PMID- 19286403 TI - Characterisation of acid-base abnormalities in pigs experimentally infected with Chlamydia suis. AB - This study characterises the acid-base abnormalities in pigs experimentally infected with Chlamydia suis (Henderson-Hasselbalch equation and Constable's simplified strong ion equation). Eight pigs were challenged with the respiratory pathogen C. suis and four pigs served as non-infected controls. Pigs were monitored from 7 days before challenge to 8 days post-inoculation. Clinical examination was performed twice daily and venous blood samples were collected every two days. Blood-gas analysis, haemoxymetry, serum biochemical analysis and electrophoresis were performed in order to characterise the acid-base derangement. Aerosol challenge with C. suis resulted in severe acid-base disturbance characterised by acute respiratory acidosis and strong ion (metabolic) acidosis secondary to anaerobic metabolism and hyper L-lactataemia. Maximal changes were seen at day 3 post-inoculation when severe clinical signs of respiratory dysfunction were evident. The results of the study provide new information regarding the pathophysiology of respiratory infection caused by C. suis and the applicability and diagnostic utility of different approaches for assessing acid-base status in pigs. PMID- 19286404 TI - Evolution of the BSE epidemic in Catalonia (1990-2015) based on a stochastic model. AB - A stochastic model was used to estimate the number of bovine spongiform encephalopathy (BSE) cases that may have occurred in Catalonia (north-east Spain) from January 1990 to December 2015. The model allowed the evolution of the unobserved cases to be estimated differentiating the population into two subgroups: (1) livestock slaughtered for human consumption without clinical signs and (2) cattle at increased risk (including fallen stock, clinical suspects, emergency slaughtered cattle and cattle with clinical signs at ante mortem inspection). If the first bovine affected by BSE had been introduced into the region in 1986, the model predicted that during the period of study the surveillance system would detect a median of 197 outbreaks (2.5-97.5 percentiles: 157-245). The estimated median number of outbreaks that may have appeared before the establishment of the monitoring program was 80 (2.5-97.5 percentiles: 35 152), and the number of cases would have peaked in 2001. PMID- 19286405 TI - Applying linear interaction energy method for binding affinity calculations of podophyllotoxin analogues with tubulin using continuum solvent model and prediction of cytotoxic activity. AB - Podophyllotoxin and its analogues have important therapeutic value in the treatment of cancer, due to their ability to induce apoptosis in cancer cells in a proliferation-independent manner. These ligands bind to colchicine binding site of tubulin near the alpha- and beta-tubulin interface and interfere with tubulin polymerization. The binding free energies of podophyllotoxin-based inhibitors of tubulin were computed using a linear interaction energy (LIE) method with a surface generalized Born (SGB) continuum solvation model. A training set of 76 podophyllotoxin analogues was used to build a binding affinity model for estimating the free energy of binding for 36 inhibitors (test set) with diverse structural modifications. The average root mean square error (RMSE) between the experimental and predicted binding free energy values was 0.56kcal/mol which is comparable to the level of accuracy achieved by the most accurate methods, such as free energy perturbation (FEP) or thermodynamic integration (TI). The squared correlation coefficient between experimental and SGB-LIE estimates for the free energy for the test set compounds is also significant (R(2)=0.733). On the basis of the analysis of the binding energy, we propose that the three-dimensional conformation of the A, B, C and D rings is important for interaction with tubulin. On the basis of this insight, 12 analogues of varying ring modification were taken, tested with LIE methodology and then validated with their experimental potencies of tubulin polymerization inhibition. Low levels of RMSE for the majority of inhibitors establish the structure-based LIE method as an efficient tool for generating more potent and specific inhibitors of tubulin by testing rationally designed lead compounds based on podophyllotoxin derivatization. PMID- 19286406 TI - VDAC-cored CaCC, CF patients best friend - finding novel therapeutics for CF. PMID- 19286407 TI - Cost-effectiveness of the modifications in the quality assurance system in radiotherapy in the example of in-vivo dosimetry. AB - PURPOSE: To present the methodology for the evaluation of cost-effectiveness of the quality assurance protocol modifications associated with increasing demands on accuracy and reliability in radiotherapy and to present results on cost effectiveness of in-vivo dosimetry as the chosen example of a technical procedure. MATERIAL AND METHODS: In-vivo dosimetry was used as an example of a quality assurance procedure, whose modifications have an impact on several procedures in the QA system and thus on the cost of radiotherapy. An analysis of 6864 patients, treated between 2001 and 2005 for tumours in the head and neck, breast, pelvis, or lung, was performed. The quality of radiotherapy was expressed as the accuracy of dose delivery and the cost was estimated from labour, equipment and materials. RESULTS: Modifications implemented in the quality assurance protocol have gradually improved the quality of irradiation. Mean deviations between measured and calculated doses, recorded for several groups of treatment sites, were reduced from -1.5% to 0.5%, 3.4% to 1.4%, 3.9% to 0.1% and 2.1% to 1.8% for head and neck, breast, pelvis and lung respectively. The standard deviations of the measured values decreased also consistently. Total monthly cost in radiotherapy (related to in-vivo dosimetry) increased from euro 4376 to euro 10,696 while the unitary cost of radiotherapy procedures remained at the same level. The predominant cost component of in-vivo dosimetry was labour, limited at first to physics staff and later extended to quality assurance personnel and technicians. CONCLUSION: The application of the presented methodology revealed cost-effectiveness relationships in tested technical procedures. PMID- 19286408 TI - Automated peritoneal dialysis-associated peritonitis due to Haemophilus influenzae showing the BLNAR phenotype. AB - A rare case of peritonitis due to beta-lactamase-negative, ampicillin-resistant Haemophilus influenzae is described in an 18-year-old male undergoing automated peritoneal dialysis. The infection was probably of respiratory origin. Two strains of H. influenzae cultured from peritoneal fluid and a nasal swab were indistinguishable by molecular methods. The patient was successfully treated with oral ciprofloxacin. The authors suggest that this species should be taken into account as the etiologic agent of peritoneal dialysis-associated peritonitis. Targeted antimicrobial therapy should be based on local antibiotic resistance patterns. PMID- 19286409 TI - Polyarticular septic arthritis due to Moraxella canis revealing multiple myeloma. PMID- 19286410 TI - High frequency of ultrasonographic effusion in interphalangeal joints of healthy subjects: a descriptive study. AB - OBJECTIVE: To describe echographic abnormalities on the proximal (PIP) and distal (DIP) interphalangeal joints of healthy subjects. METHODS: Healthy asymptomatic volunteers under forty were enrolled. Ultrasonography was performed on PIP and DIP 2-5 in the presence of two operators. In B mode, synovial hypertophy and effusion were defined with the OMERACT criteria. In power Doppler mode, the inflammatory activity was evaluated. RESULTS: 46 subjects were enrolled: mean age 25.5 years, 89% right handed. 368 PIPs and DIPs were scanned. Effusion was found in 19.6% of dominant PIPs versus 22.3% of non-dominant PIPs (p>0.05). Effusion could be uni- or bilateral. DIP effusion was less frequent (3% DIPs). Average size of effusion was 0.83+/-0.25 mm for PIP and 0.72+/-0.16 mm for DIP. CONCLUSION: Our study proposes a descriptive analysis of the PIP and the DIP joints. It confirms, with a bigger number of studied joints, the possible existence of a physiological effusion of the PIP of healthy subjects. It is the first descriptive study of the normal DIP where the presence of an effusion is very rare. PMID- 19286411 TI - [How I do... to realize a spatio-temporal-imaging-correlation (STIC)]. PMID- 19286412 TI - Gravidity complicated by abdominal neoplastic process. AB - The case of 31-year-old pregnant woman in the 28th week of pregnancy is presented. She was brought in a bad condition to a small hospital by her parents. The case history of only 5 h included e.g. nausea, multiple emesis, cephalea, deteriorated respiration. Shock status was diagnosed in the hospital intensive care unit. After the patient lost her consciousness, resuscitation, intubation and artificial ventilation breath control were realised immediately, the doses of 13 mg of adrenalin were applied. PMID- 19286413 TI - Relevance of collagen piezoelectricity to "Wolff's Law": a critical review. AB - According to "Wolff's Law", bone is deposited and reinforced at areas of greatest stress. From a clinical perspective, this "law" is supported by the strong association between bone density and physical activity. From a mechanistic standpoint, however, the law presents a challenge to scientists seeking to understand how osteocytes and osteoblasts sense the mechanical load. In the 1960s, collagen piezoelectricity was invoked as a potential mechanism by which osteocytes could detect areas of greater stress but piezoelectricity diminished in importance as more compelling mechanisms, such as streaming potential, were identified. In addition, accumulating evidence for the role of fluid-related shear stress in osteocyte's mechanosensory function has made piezoelectricity seemingly more obsolete in bone physiology. This review critically evaluates the role of collagen piezoelectricity (if any) in Wolff's Law--specifically, the evidence regarding its involvement in strain-generated potentials, existing alternate mechanisms, the present understanding of bone mechanosensation, and whether piezoelectricity serves an influential role within the context of this newly proposed mechanism. In addition to reviewing the literature, this review generates several hypotheses and proposes future research to fully address the relevance of piezoelectricity in bone physiology. PMID- 19286414 TI - New insights into rhythmic brain activity from TMS-EEG studies. AB - There is renewed interest in the functional role of oscillatory brain activity in specific frequency bands, investigated in humans through electroencephalography (EEG) and magnetoencephalography (MEG) recordings. In parallel, there is a growing body of research on non-invasive direct stimulation of the human brain via repetitive (rhythmic) transcranial magnetic stimulation (TMS), and on those frequencies that have the strongest behavioural impact. There is, therefore, great potential in combining these two lines of research to foster knowledge on brain rhythms, in addition to potential therapeutic applications of rhythmic brain stimulation. Here, we review findings from this rapidly evolving field linking intrinsic brain oscillations to distinct sensory, motor and cognitive operations. The findings emphasize that brain rhythms are causally implicated in cognitive functions. PMID- 19286415 TI - Copper-dioxygen complex mediated C-H bond oxygenation: relevance for particulate methane monooxygenase (pMMO). AB - Particulate methane monooxygenase (pMMO), an integral membrane protein found in methanotrophic bacteria, catalyzes the oxidation of methane to methanol. Expression and greater activity of the enzyme in the presence of copper ion suggest that pMMO is a cuprous metalloenzyme. Recent advances - especially the first crystal structures of pMMO - have energized the field, but the nature of the active site(s) and the mechanism of methane oxidation remain poorly understood-yet hotly contested. Herein the authors briefly review the current understanding of the pMMO metal sites and discuss advances in small molecule Cu O(2) chemistry that may contribute to an understanding of copper-ion mediated hydrocarbon oxidation chemistry. PMID- 19286416 TI - EPR studies of VO2+ doped bis(saccharinato)bis(pyridine) zinc (II) single crystals. AB - Electron paramagnetic resonance (EPR) studies of vanadyl ions, VO2+, as paramagnetic impurity in bis(saccharinato)bis(pyridine)zinc (II) [hereafter, Zn(sac)2(py)2] are made at room temperature. The angular variation of the EPR spectra has shown that three different VO2+ complexes are located in different chemical environments and each environment contains two magnetically inequivalent VO2+ sites in distinct orientations occupying substitutional positions in the lattice and show very high angular dependence. The spin-Hamiltonian parameters are determined and these parameters have been used to assess the bonding coefficients of the VO2+ ion in [Zn(sac)2(py)2] lattice. The values of spin Hamiltonian parameters indicate that the VO2+ ions in [Zn(sac)2(py)2] were present in octahedral coordination with a tetragonal compression and belong to C4v symmetry. The parallel and perpendicular components of axially symmetric g and hyperfine tensors are evaluated and the results are discussed. PMID- 19286417 TI - Spectroscopic studies of interactions between C.I. Reactive Orange 16 with alkyltrimethylammonium bromide surfactants. AB - In the present study, the influence of surfactants on spectral properties of an azo dye in aqueous solutions has been investigated by means of UV-vis spectroscopy in submicellar and micellar concentration range. The spectral signature of the polarity of the azo dye C.I. Reactive Orange 16 (RO16) exhibits sensitivity to the polarity of the dye's environment. This dependence of absorption on microenvironment was used to investigate the ion pair complex formed from electrostatic interaction of a series of alkyltrimethylammonium bromide surfactants (CmTAB, m=12, 14, 16 and 18) with the anionic azo dye RO16. It was observed that the aggregation of surfactant and dye takes place at surfactant concentration far below the critical micelle concentration of the individual surfactant. Aggregation is reflected by the appearance of a new absorption band in the spectrum of the dye. Spectral behavior of dye-surfactant solution with varying concentration of surfactant confirms that electrostatic interaction between dye and surfactant occurs up to a certain level. Beyond this concentration, with addition of surfactant, micelles occur and all dye molecules are accommodated into a normal micelle as monomeric molecules. The short-range hydro phobic interactions are very important factors as the long-range electrostatic forces on the dye-surfactant aggregation in aqueous solution. The effect of the length of the alkyl chain of the surfactant on the complex formation between cationic surfactant and reactive dye was that the hydrophobicity of alkyl chains plays an important role in complex formation. Going from less hydrophobic solution to the more hydrophobic micellar environment, the occurrence of complex is found at lower surfactant concentration. Because the CMC values of dye-surfactant solution are decreased with increasing alkyl chain length. PMID- 19286418 TI - Copper(II) complexes with 2-pyridineformamide-derived thiosemicarbazones: spectral studies and toxicity against Artemia salina. AB - The copper(II) complexes [Cu(H2Am4DH)Cl2] (1), [Cu(H2Am4Me)Cl2] (2), [Cu(H2Am4Et)Cl2] (3) and [Cu(2Am4Ph)Cl] (4) with 2-pyridineformamide thiosemicarbazone (H2Am4DH) and its N(4)-methyl (H2Am4Me), N(4)-ethyl (H2Am4Et) and N(4)-phenyl (H2Am4Ph) derivatives were studied by means of infrared and EPR spectral techniques. The crystal structure of 4 was determined. The studied compounds proved to be toxic to Artemia salina, suggesting that they could present cytotoxic activity against solid tumors. Among the free thiosemicarbazones H2Am4Ph presented higher toxicity than all other compounds, which showed comparable effects. In the case of complexes 2 and 3 toxicity is probably attributable to the complex as an entity or to a synergistic effect involving the thiosemicarbazone and copper. H2Am4Ph and complexes 2 and 3 revealed to be the most promising compounds as potential antineoplasic agents. PMID- 19286419 TI - Performance evaluation of the QIAGEN EZ1 DSP Virus Kit with Abbott RealTime HIV 1, HBV and HCV assays. AB - BACKGROUND: Automated sample preparation systems must meet the demands of routine diagnostics laboratories with regard to performance characteristics and compatibility with downstream assays. OBJECTIVES: In this study, the performance of QIAGEN EZ1 DSP Virus Kit on the BioRobot EZ1 DSP was evaluated in combination with the Abbott RealTime HIV-1, HCV, and HBV assays, followed by thermalcycling and detection on the Abbott m2000rt platform. STUDY DESIGN: The following performance characteristics were evaluated: linear range and precision, sensitivity, cross-contamination, effects of interfering substances and correlation. RESULTS: Linearity was observed within the tested ranges (for HIV-1: 2.0-6.0 log copies/ml, HCV: 1.3-6.9 log IU/ml, HBV: 1.6-7.6 log copies/ml). Excellent precision was obtained (inter-assay standard deviation for HIV-1: 0.06 0.17 log copies/ml (>2.17 log copies/ml), HCV: 0.05-0.11 log IU/ml (>2.09 log IU/ml), HBV: 0.03-0.07 log copies/ml (>2.55 log copies/ml)), with good sensitivity (95% hit rates for HIV-1: 50 copies/ml, HCV: 12.5 IU/ml, HBV: 10 IU/ml). No cross-contamination was observed, as well as no negative impact of elevated levels of various interfering substances. In addition, HCV and HBV viral load measurements after BioRobot EZ1 DSP extraction correlated well with those obtained after Abbott m2000sp extraction. CONCLUSIONS: This evaluation demonstrates that the QIAGEN EZ1 DSP Virus Kit provides an attractive solution for fully automated, low throughput sample preparation for use with the Abbott RealTime HIV-1, HCV, and HBV assays. PMID- 19286420 TI - Identification of cognitive deficits in amyotrophic lateral sclerosis. PMID- 19286421 TI - Osmo-therapy for seizures: a new treatment paradigm? PMID- 19286422 TI - Quality of life in patients with locally advanced prostate cancer given endocrine treatment with or without radiotherapy: 4-year follow-up of SPCG-7/SFUO-3, an open-label, randomised, phase III trial. AB - BACKGROUND: Androgen treatment for prostate cancer can adversely affect functional domains of quality of life. We aimed to assess quality of life in men with locally advanced prostate cancer in an open-label phase III randomised comparison between lifelong endocrine treatment with and without radiotherapy. METHODS: We obtained quality-of-life information from 872 (99%) of 875 eligible men with locally advanced prostate cancer (T3; 78%) who were randomly assigned, between 1996 and 2002, to 3 months of total androgen blockade followed by continuous endocrine treatment (439 patients) or the same hormonal treatment with radiotherapy 3 months after randomisation (436 patients). Prospective outcomes included patient-reported symptoms and quality of life assessed with questionnaires from baseline to 4 years after randomisation. Analysis was by intention to treat. This study is registered as an international standard randomised controlled trial, number ISRCTN01534787. FINDINGS: 438 of 439 men assigned endocrine treatment and 434 of 436 assigned endocrine plus radiotherapy completed at least one questionnaire. Missing data at baseline and during follow up was equally distributed between groups. At 4 years, 64 (18%) of 353 patients on combined therapy and 39 (12%) of 337 on endocrine-alone therapy had moderate to severe urinary bother (p=0.005), and 16 (4%) of 355 on combined therapy and five (2%) of 338 on endocrine treatment alone had pain while urinating (p=0.024). 37 (11%) of 350 in the combined group and 23 (7%) of 35 in the endocrine-only group had overall bother from all bowel symptoms (p=0.022). 281 (85%) of 332 in the combined-treatment group and 227 (72%) of 313 in the endocrine-only group had erectile dysfunction (p=0.0002). Quality of life at 4 years was similar, with the exception of decreased social function in patients receiving endocrine treatment plus radiotherapy. INTERPRETATION: Although addition of radiotherapy to endocrine treatment significantly increased some treatment-related symptoms, none were serious. Given the substantial survival benefit of combined treatment, the increase of symptoms seems acceptable and has little extra effect on quality of life after 4 years compared with endocrine treatment alone. PMID- 19286423 TI - Inhaled muscarinic antagonists for COPD--does an anti-inflammatory mechanism really play a role? AB - The long acting inhaled muscarinic antagonist tiotropium (Spiriva) improves lung function in patients with COPD. In addition, tiotropium reduces exacerbation frequency, dyspnoea and improves exercise capacity. As the latter has been associated with airway inflammation then this suggests that, in addition to the well-known anti-bronchoconstrictor effect, tiotropium might also display anti inflammatory properties. With our current state of knowledge, however, it is not necessary to postulate an anti-inflammatory effect for tiotropium (Spiriva), rather inhibition of smooth muscle constriction with subsequent effects on lung hyperinflation (and possibly pulmonary circulation) can explain the effects on exacerbation frequency, dyspnoea and exercise capacity. Recent reports suggest that tiotropium can inhibit viral activation of inflammation and vagal nerve stimulation, suggesting a mechanism by which tiotropium can inhibit viral induction of exacerbations in COPD. PMID- 19286424 TI - Serotonin and pulmonary hypertension--from bench to bedside? AB - The serotonin hypothesis of pulmonary arterial hypertension (PAH) arose owing to anorexigens, acting as indirect serotinergic agonists, causing PAH. However, it is now thought that serotonin plays an important role in the pathobiology of PAH per se. The rate-limiting enzyme in the synthesis of peripheral serotonin is tryptophan hydroxylase 1 (TPH1), serotonin can mediate pulmonary arterial smooth muscle cell proliferation via the serotonin transporter (SERT) and serotonin can induce pulmonary vasoconstriction via the 5-HT1B receptor in man. There is evidence that TPH1, SERT and 5-HT1B expression/activity can be upregulated in clinical PAH. This review discusses recent evidence implicating serotonin in the development of experimental and clinical PAH and suggests potential therapeutic targets. PMID- 19286425 TI - Lymphocyte kinetics in health and disease. AB - Quantitative understanding of immunology requires the development of experimental and mathematical techniques for estimation of rates of division and death of lymphocytes under different conditions. Here, we review the advantages and limitations of several labelling methods that are currently used to quantify turnover of lymphocytes in vivo. In addition to highlighting insights into lymphocyte kinetics which have recently been gained thanks to the development of novel techniques, we discuss important directions for future experimental and theoretical work in the field of lymphocyte turnover. PMID- 19286426 TI - Do we have memory of danger as well as antigen? AB - Recognition of specific protein antigens leads to immunological memory of antigen, whereas recognition of danger signals by the innate immune system determines the size, nature and longevity of a response. Recent data indicate that recognition of danger might have long-lasting effects on CD8 memory T-cell populations, specifically enhancing early cytokine release and thus altering the nature of subsequent immune responses. Here, a modified model of immune regulation is proposed in which a nonspecific immunological memory of danger accumulates during maturation of the immune system, allowing more potent cell mediated immunity to develop in dangerous environments. Such a mechanism could be involved in phenomena leading to the 'hygiene hypothesis'. PMID- 19286427 TI - Simple and rapid liquid chromatography method for determination of moxifloxacin in plasma. AB - A high performance liquid chromatographic method for determination of moxifloxacin in human plasma was developed. The method involved deproteinisation of the sample with perchloric acid and analysis of the supernatant using a reversed-phase C(18) column (150 mm) and fluorescence detection at an excitation wavelength of 290 nm and an emission wavelength of 460 nm. The assay was specific for moxifloxacin and linear from 0.125 to 10.0 microg/ml. The relative standard deviation of intra- and inter-day assays was lower than 10%. The average recovery of moxifloxacin from plasma was 101%. Due to its simplicity, the assay can be used for pharmacokinetic studies of moxifloxacin. PMID- 19286428 TI - High-throughput shotgun lipidomics by quadrupole time-of-flight mass spectrometry. AB - Technological advances in mass spectrometry and meticulous method development have produced several shotgun lipidomic approaches capable of characterizing lipid species by direct analysis of total lipid extracts. Shotgun lipidomics by hybrid quadrupole time-of-flight mass spectrometry allows the absolute quantification of hundreds of molecular glycerophospholipid species, glycerolipid species, sphingolipid species and sterol lipids. Future applications in clinical cohort studies demand detailed lipid molecule information and the application of high-throughput lipidomics platforms. In this review we describe a novel high throughput shotgun lipidomic platform based on 96-well robot-assisted lipid extraction, automated sample infusion by mircofluidic-based nanoelectrospray ionization, and quantitative multiple precursor ion scanning analysis on a quadrupole time-of-flight mass spectrometer. Using this platform to compile comprehensive lipid arrays associated with metabolic dysfunctions is a powerful strategy for pinpointing the mechanistic details by which alterations in tissue specific lipid metabolism are directly linked to the etiology of many lipid mediated disorders. PMID- 19286429 TI - Displacement effect during HPLC preparative purification of human insulin. AB - HPLC plays a key role in the preparative purification of human insulin. A21 desamidoinsulin is one of the impurities that possesses the chromatographic behavior similar to that of insulin and hence separation from this by-product is rather difficult at the process scale. During the optimization of insulin reversed-phase HPLC purification, when a column was sufficiently overloaded, the effect of displacement of A21-desamidoinsulin molecules from active groups of sorbent by insulin ones was observed. It was suggested that monocarboxylic acid and organic modifier in mobile phase are responsible for the esterification during which the formed ester promotes the displacement effect. This effect was studied in order to optimize the purification of human insulin at the process scale. PMID- 19286430 TI - Determination of pyrrole-imidazole polyamide in rat plasma by liquid chromatography-tandem mass spectrometry. AB - Pyrrole (Py)-imidazole (Im) polyamides synthesized by combining N-methylpyrrole and N-methylimidazole amino acids have been identified as novel candidates for gene therapy. In this study, a sensitive method using liquid chromatography tandem mass spectrometry (LC-MS/MS) with an electrospray ionization (ESI) source was developed and validated for the determination and quantification of Py-Im polyamide in rat plasma. Py-Im polyamide was extracted from rat plasma by solid phase extraction (SPE) using a Waters Oasis HLB cartridge. Separation was achieved on an ACQUITY UPLC HSS T3 (1.8 microm, 2.1 x 50 mm) column by gradient elution using acetonitrile:distilled water:acetic acid (5:95:0.1, v/v/v) and acetonitrile:distilled water:acetic acid (95:5:0.1, v/v/v). The method was validated over the range of 10-1000 ng/mL and the lower limit of quantification (LLOQ) was 10 ng/mL. This method was successfully applied to the investigation of the pharmacokinetics of Py-Im polyamide after intravenous administration. PMID- 19286431 TI - Polysaccharide surface modified Fe3O4 nanoparticles for camptothecin loading and release. AB - Fe(3)O(4) nanoparticles were stabilized using different functional polysaccharides, such as chitosan (CS), O-carboxymethylchitosan (OCMCS) and (N succinyl-O-carboxymethylchitosan (NSOCMCS) to improve their bioactivity. The release profile and the in vitro cancer cell inhibition activity of camptothecin (CPT) loaded polysaccharide modified Fe(3)O(4) nanoparticles were systematically studies. The particle size and size distribution of CPT-loaded polysaccharide modified Fe(3)O(4) nanoparticles were found to be strongly dependent on polysaccharide character. Such polysaccharide character could also affect CPT adsorption efficiency, CPT release behavior and bovine serum albumin (BSA) unspecific binding capacity. After 24 h incubation of 7721 cancer cells with CPT loaded polysaccharide modified Fe(3)O(4) nanoparticles, significant changes in cell morphology could be discernible from phase contrast microscopy. Cytotoxicity assay showed these polysaccharide modified Fe(3)O(4) nanoparticles did not exhibit noteworthy cytotoxicity against 7721, however, the in vitro inhibition rate of CPT-loaded polysaccharide modified Fe(3)O(4) nanoparticles against 7721 liver cancer cell increased significantly in comparison with that of CPT-free drug. PMID- 19286432 TI - Ocular injectable formulation assessment for oxidized dextran-based hydrogels. AB - Initiator-free injectable hydrogels are very interesting for drug and/or cell delivery applications, since they can be administered in a minimally invasive way, and avoid the use of potentially harmful chemical initiators. In the current work, oxidized dextran crosslinked with adipic acid dihydrazide hydrogels were further characterized and tuned to produce formulations, with the aim of producing an injectable formulation for the possible treatment of posterior eye diseases. The gelation rate and the hydrogel dissolution profile were shown to be dependent on the balance between the degree of dextran oxidation, and the concentration of both components. For the in vitro studies, rabbit corneal endothelial cells were seeded on the hydrogels to assess cytotoxicity. Hydrogels prepared with low oxidized dextrans were able to promote cell adhesion and proliferation to confluence in just 24h, while more highly oxidized samples promoted cell adhesion and proliferation, but without achieving confluence. Cell viability studies were performed using MTS assays to verify the non-cytotoxicity of hydrogels and their degradation byproducts, rendering these formulations attractive for further in vivo studies. PMID- 19286433 TI - In vivo biocompatibility and vascularization of biodegradable porous polyurethane scaffolds for tissue engineering. AB - Scaffolds for tissue engineering should be biocompatible and stimulate rapid blood vessel ingrowth. Herein, we analyzed in vivo the biocompatibility and vascularization of three novel types of biodegradable porous polyurethane scaffolds. The polyurethane scaffolds, i.e., PU-S, PU-M and PU-F, were implanted into dorsal skinfold chambers of BALB/c mice. Using intravital fluorescence microscopy we analyzed vascularization of the implants and venular leukocyte endothelial cell interaction in the surrounding host tissue over a 14 day period. Incorporation of the scaffolds was analyzed by histology, and a WST-1 assay was performed to evaluate their cell biocompatibility in vitro. Our results indicate that none of the polyurethane scaffolds was cytotoxic. Accordingly, rolling and adherent leukocytes in venules of the dorsal skinfold chamber were found in a physiological range after scaffold implantation and did not significantly differ between the groups, indicating a good in vivo biocompatibility. However, the three scaffolds induced a weak angiogenic response with a microvessel density of only approximately 47-60 and approximately 3-10cm/cm(2) in the border and centre zones of the scaffolds at day 14 after implantation. Histology demonstrated that the scaffolds were incorporated in a granulation tissue, which exhibited only a few blood vessels and inflammatory cells. In conclusion, PU-S, PU-M and PU-F scaffolds may be used to generate tissue constructs which do not induce a strong inflammatory reaction after implantation into patients. However, the scaffolds should be further modified or conditioned in order to accelerate and improve the process of vascularization. PMID- 19286434 TI - Decellularization of bovine corneas for tissue engineering applications. AB - Scaffolds derived from processed tissues offer viable alternatives to synthetic polymers as biological scaffolds for regenerative medicine. Tissue-derived scaffolds provide an extracellular matrix (ECM) as the starting material for wound healing and the functional reconstruction of tissues, offering a potentially valuable approach for the replacement of damaged or missing tissues. Additionally, acellular tissue may provide a natural microenvironment for host cell migration and the induction of stem cell differentiation to contribute to tissue regeneration. There are a number of processing methods that aim to stabilize and provide an immunologically inert tissue scaffold. Furthermore, these tissue-processing methods can often be applied to xenogenic transplants because the essential components of the ECM are often maintained between species. In this study, we applied several tissue-processing protocols to the cornea in order to obtain a decellularized cornea matrix that maintained the clarity and mechanical properties of the native tissue. Histology, mechanical testing and electron microscopy techniques were used to assess the cell extraction process and the organization of the remaining ECM. In vitro cell seeding experiments confirmed the processed corneas' biocompatibility. PMID- 19286435 TI - Preparation of nanometer-scale rod array of hydroxyapatite crystal. AB - Fabrication of nano- or micro-structured scaffolds to mimic structural and three dimensional details of natural bone or teeth has been the subject of much interest, and this study proposes a new strategy for self-assembling one dimensional hydroxyapatite (HAp) nanorods into organized superstructures. A nanometer-scale rod array of HAp having preferred orientation to the c-axis was successfully prepared simply by soaking calcium-containing silicate glass substrates in Na(2)HPO(4) aqueous solution at 80 degrees C for various periods. Those HAp rods grew perpendicularly to the glass surface, and the crystallites covered the glass surface uniformly, resulting in a "dental enamel-like" rod array structure consisting of "pine-leaf-like" structure units. PMID- 19286436 TI - A simple nasal-packing technique for minor nasal surgery which reduces pain associated with removal of packs: a surgical technique. PMID- 19286437 TI - Giant naevus, giant excision, eleg(i)ant closure? Reconstructive surgery with Integra Artificial Skin to treat giant congenital melanocytic naevi in children. AB - BACKGROUND: The optimal surgical treatment of giant congenital melanocytic naevi remains a considerable challenge in reconstructive surgery. None of the currently available techniques is universally applicable. The goal of this pilot study was to test Integra Artificial Skin (Integra) in the surgical treatment of giant congenital melanocytic naevi. METHODS: Between May 2000 and March 2004, Integra was used in 12 children (n=12; aged seven months to 11 years, mean 3.8 years). Giant congenital melanocytic naevi covered 1-12% of the total body surface area (mean 4.2%) located over the trunk in 50%, and over face and extremities in 25% each. RESULTS: In eight children, Integra implantation was primarily successful; in four patients a partial or complete removal and re-implantation was necessary due to complications. The final take rate of Integra ranged from 95-100%, except for one patient with a take rate of 30% (mean 93%). Second stage split-thickness skin grafting yielded take rates from 95-100% (mean 98%). Functional and cosmetic outcome was rated excellent in 58%, good in 25% and fair in 17% (follow-up six months to four years, mean 2.2 years). CONCLUSIONS: These results suggest that Integra is a new and valid method to successfully treat giant congenital melanocytic naevi in early childhood in a definitive manner and with high-quality results. PMID- 19286438 TI - The deepithelialisation technique for monitoring free flaps in heavily pigmented skin. PMID- 19286439 TI - A twitching free muscle flap: causes and implications. PMID- 19286440 TI - Reconstruction for sternal osteomyelitis at the lower third of sternum. AB - BACKGROUND: Sternal wound infection causes considerable morbidity and mortality for open-heart patients. Treatment of the wounds at the upper two-thirds is easier with pectoralis major muscle or other flaps. However, there would be more problems with the lower one-third sternal wounds. METHODS: From 1983 to 2007, 32 patients of osteomyelitis involving the lower sternum were treated with one of the following methods: (1) Latissimus dorsi with fasciocutaneous extension flap (2) Tri-pedicled pectoralis major musculocutaneous flap (3) Pectoralis major muscle with rectus abdominis muscle flap (4) Pectoralis major muscle with omentum flap (5) Free vastus lateralis muscle flap and skin grafting RESULTS: The viability of these flaps was good except for one of the five patients with pectoralis major-rectus abdominis muscle. One of the patients from the free vastus lateralis muscle group died of heart failure 6 weeks after surgery, but the coverage of sternal wound was successful. No recurrent sternal infection was found. CONCLUSIONS: For coverage of sternal wounds, the transferred tissue must have optimal blood supply in order to overcome the infection. According to the descending degree of ease, the ladder of reconstruction is from (1) to (5), depending on the relative length of the sternal wound and the arc of rotation of these flaps. In pectoralis major with rectus abdominis flap group, it is suggested that the upper sternal wound be covered with pectoralis major muscle but lower third sternal wounds with omentum instead of rectus abdominis muscle. PMID- 19286441 TI - Two challenges (amongst the many). PMID- 19286442 TI - Platelet glycoprotein IIb/IIIa (HPA-1 and HPA-3) polymorphisms in patients with hemorrhagic fever with renal syndrome. AB - This study investigated the relationship between human platelet alloantigen (HPA) polymorphisms of glycoprotein IIb/IIIa and hemorrhagic fever with renal syndrome (HFRS). HPA-1 and HPA-3 genotyping was performed with allele-specific primer polymerase chain reaction in 104 patients with HFRS and 100 normal individuals as controls. The relationships between gene polymorphisms of HPAs and HFRS and the disease severity were analyzed. The results indicated no significant difference in HPA-1 genotype distributions (p > 0.05), but a significant difference in the distributions of genotype and allele frequencies of HPA-3 between HFRS patients and controls (p < 0.01). The distributions of HPA-3 genotype and allele frequencies differed significantly between patients with different clinical types and the HPA-3 b allele was more frequently observed in patients with more severe clinical types. These results indicate that HPA-3 polymorphism may be one of the inherited risk factors associated with the susceptibility of hantavirus infection and the disease severity of HFRS. PMID- 19286443 TI - Prevalence of vitamin D receptor gene polymorphisms FokI and BsmI in Brazilian individuals with type 1 diabetes and their relation to beta-cell autoimmunity and to remaining beta-cell function. AB - The effect of the vitamin D receptor (VDR) gene polymorphisms on susceptibility to type 1 diabetes (T1DM) is heterogeneous. Genetic factors may also influence the residual beta-cell function. We studied the frequency of VDR FokI (rs10735810) and BsmI (rs154410) polymorphisms in T1DM and their relationship to beta-cell autoimmunity and residual beta-cell function. We genotyped 189 T1DM (diabetes duration, 7.1 +/- 5.4 years) and 194 controls (C) by restriction length polymorphism-polymerase chain reaction. GAD65Ab, IA2Ab, ionized calcium (iCa), HbA(1c)and fasting C-peptide (FCP) were evaluated. FCP values greater than 0.6 ng/ml were considered as residual beta-cell function. The BsmI was more frequent in the C (bb plus Bb 79.1 C vs. 66.1% T1DM, p = 0.006), and the FokI polymorphism frequencies were similar between T1DM and C. We did not observe differences in pancreatic autoantibody profiles according to VDR genotypes. We observed that T1DM with f allele tended to have lower residual pancreatic beta-cell function (5.8% ff and Ff vs. 14.3% FF, p = 0.074) with similar age, diabetes duration, AAb positivity, HbA(1c), and iCa. Age at diagnosis of T1DM with BsmI polymorphism tended to be greater (10.7 +/- 4.9 bb and Bb vs. 9.3 +/- 4.5 years BB, p = 0.06). In conclusion, the results of this study showed no relationship between VDR polymorphisms and beta-cell autoimmunity; however we observed a relationship with age and remaining beta-cell function in Brazilian individuals with T1DM. These data may contribute to understanding the heterogeneous relationship between genetic markers and clinical features observed in this disease. PMID- 19286444 TI - Possible impact of MADCAM1 gene single nucleotide polymorphisms to the outcome of allogeneic hematopoietic stem cell transplantation. AB - Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) contributes to the recruitment of donor T cells into the mucosal tissues of the recipient after allogeneic hematopoietic stem cell transplantation (aHSCT). The aim of our study was to determine whether selected single nucleotide polymorphisms (SNPs) of the MADCAM1 gene are associated with development of serious complications after aHSCT. Three MADCAM1 gene single nucleotide polymorphisms (rs758502 C/T, rs2302217 A/G, rs3745925 G/T) were genotyped by polymerase chain reaction with sequence-specific primers in 87 Czech, HLA-identical donor-recipient aHSCT pairs. MADCAM1 rs2302217 AA homozygous recipients developed chronic GVHD more frequently than patients with other genotypes (65% vs. 34%; p = 0.025). Furthermore, multivariate analysis revealed the MADCAM1 rs2302217 AA genotype in recipient being also an independent factor associated with development of acute GVHD (p = 0.036) and decreased overall survival (p = 0.001). These data suggest that MADCAM1 gene polymorphisms may be associated with the risk of chronic GVHD and may, also, affect mortality related to aHSCT. PMID- 19286445 TI - Additional sequence analysis outside exon 2 clarifies DRB1*12 and DRB1*14 allelic frequencies in Koreans. AB - Polymorphisms outside exon 2 of the DRB1 coding region exhibit increased ambiguities in allele assignments in typing laboratories analyzing the exon 2 sequence alone. This study investigated the frequencies of alleles in DRB1*120101/1206/1210 and DRB1*140101/1454 groups by DNA sequencing of exons 1 and/or 3 and examined their associations with class II alleles at adjacent loci in Koreans. Of 103 individuals, 97 were DRB1*120101 and 6 were DRB1*1210. None carried DRB1*1206, an allele initially characterized from a Korean individual. Association of DRB1*120101 with various DRB3, DQA1, and DQB1 alleles was reflected in 11 different four-locus haplotypes. In contrast, DRB1*1210 was exclusively associated with DRB3*020201, DQA1*0505, and DQB1*0301. All 97 DRB1*140101/1454 samples were DRB1*1454. DRB1*1454 was strongly associated with DRB3*020201, DQA1*010401, and two different DQB1 alleles (*0502, *0503). Continuous updating of HLA high-resolution typing information will be useful in the development of typing and matching strategies for hematopoietic stem cell transplantation. PMID- 19286446 TI - Antidepressant-like effect of the extract of Rosmarinus officinalis in mice: involvement of the monoaminergic system. AB - Rosemary, Rosmarinus officinalis L. (Labiatae) has several therapeutic applications in folk medicine in curing or managing a wide range of diseases, including depression. In this study, the effect of the hydroalcoholic extract of the stems and leaves of this plant was investigated in two behavioral models, the forced swimming test (FST) and tail suspension test (TST) in mice. The extract of R. officinalis produced an antidepressant-like effect, since the acute treatment of mice with the extract by p.o. route significantly reduced the immobility time in the FST (100 mg/kg) and TST (10-100 mg/kg), as compared to a control group, without accompanying changes in ambulation in the open-field test. Moreover, the repeated administration (14 days) of the hydroalcoholic extract of R. officinalis by p.o. route also produced an antidepressant-like effect in the TST (100-300 mg/kg). The pretreatment of mice with p-chlorophenylalanine (PCPA, 100 mg/kg, i.p., an inhibitor of serotonin synthesis, for 4 consecutive days), NAN-190 (0.5 mg/kg, i.p., a 5-HT(1A) receptor antagonist), ketanserin (5 mg/kg, i.p., a 5 HT(2A) receptor antagonist), 1-(m-chlorophenyl) biguanide (mCPBG, 10 mg/kg, i.p., a 5-HT(3) receptor agonist), prazosin (1 mg/kg, i.p., an alpha(1-)adrenoceptor antagonist), SCH23390 (0.05 mg/kg, s.c., a dopamine D(1) receptor antagonist) or sulpiride (50 mg/kg, i.p., a dopamine D(2) receptor antagonist), but not yohimbine (1 mg/kg, i.p., an alpha(2-)adrenoceptor antagonist) was able to reverse the anti-immobility effect of the extract (10 mg/kg, p.o.) in the TST. The combination of MDL72222, (0.1 mg/kg, i.p., a 5-HT(3) receptor antagonist) with a sub-effective dose of the extract of R. officinalis (1 mg/kg, p.o.) produced an anti-immobility effect in the TST. The results suggest that the antidepressant action of the extract of R. officinalis is mediated by an interaction with the monoaminergic system and that this plant should be further investigated as an alternative therapeutic approach for the treatment of depression. PMID- 19286447 TI - Studies on protein characteristics and toxic constituents of Simarouba glauca oilseed meal. AB - In order to exploit the protein rich (47.7 g/100g) simarouba meal in food/feed, studies were conducted on its chemical composition with emphasis on protein characteristics and toxic constituents. Simarouba meal contained high calcium (143 mg/100g) and sodium (79 mg/100g). Saponins with triterpenoid aglycone (3.7 g/100g), alkaloids (1.01 g/100g), phenolics (0.95 g/100g) and phytic acid (0.73 g/100g) were the major toxic constituents identified in simarouba meal. TLC and HPLC results indicated that among different fractions of simarouba saponins, one dominant fraction accounted for about 28%. Proteins of simarouba recorded high in vitro digestibility (88%). SDS-PAGE revealed four major protein bands in molecular weight ranges of 20-24, 36-45 and 55-66 kDa. Apart from, glutamic acid (23.43 g/100g protein) and arginine (10.75 g/100g protein), simarouba protein contained high essential amino acids like leucine (7.76 g/100g protein), lysine (5.62 g/100g protein) and valine (6.12 g/100g protein). Among nutritional indices, simarouba meal recorded a good EAA Index (75.02), C-PER (1.90) and PDCAAS (1.0-Adult group). PMID- 19286448 TI - Determination hematotoxic and hepatotoxic effects of trichloroacetic acid at sublethal dosage in rats. AB - The present study was designed to understand the effects of sublethal concentration of trichloroacetic acid (TCA) on hematological constituents [Red Blood Corpuscles (RBC) White Blood Corpuscles (WBC), Mean Cell Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC), Platelet (PLT) counts, Hemoglobin (HB) and Hematocrite (HCT) levels] and liver damage biomarker [cholesterol (CHL), protein (PRT) and bilirubin (BIL)] in rats at subchronic period under laboratory conditions. 2000 ppm dosage of TCA was administered orally to 6 male rats ad libitum during the tests for 52 days consecutively. Results showed that TCA caused a significant increase in the serum BIL whereas decrease in PRT and CHL. With regards to hematological constituents, while RBC, MCV, MCH, MCHC, HB and HCT level decreased significantly, PLT counts increased in rats treated with 2000 ppm of TCA exposure. The observations presented led us to conclude that TCA administration of subchronic at sublethal dosage induced the liver damage and possessed hematotoxic in rat. These data, along with the determined changes suggest that TCA produce substantial systemic organ toxicity in rats during the period of a 52-day subchronic exposure. PMID- 19286449 TI - A semi-quantitative model for risk appreciation and risk weighing. AB - Risk managers need detailed information on (1) the type of effect, (2) the size (severity) of the expected effect(s) and (3) the fraction of the population at risk to decide on well-balanced risk reduction measures. A previously developed integrated probabilistic risk assessment (IPRA) model provides quantitative information on these three parameters. A semi-quantitative tool is presented that combines information on these parameters into easy-readable charts that will facilitate risk evaluations of exposure situations and decisions on risk reduction measures. This tool is based on a concept of health impact categorization that has been successfully in force for several years within several emergency planning programs. Four health impact categories are distinguished: No-Health Impact, Low-Health Impact, Moderate-Health Impact and Severe-Health Impact. Two different charts are presented to graphically present the information on the three parameters of interest. A bar plot provides an overview of all health effects involved, including information on the fraction of the exposed population in each of the four health impact categories. Secondly, a Health Impact Chart is presented to provide more detailed information on the estimated health impact in a given exposure situation. These graphs will facilitate the discussions on appropriate risk reduction measures to be taken. PMID- 19286450 TI - Probabilistic modelling of exposure doses and implications for health risk characterization: glycoalkaloids from potatoes. AB - Potatoes are a source of glycoalkaloids (GAs) represented primarily by alpha solanine and alpha-chaconine (about 95%). Content of GAs in tubers is usually 10 100 mg/kg and maximum levels do not exceed 200 mg/kg. GAs can be hazardous for human health. Poisoning involve gastrointestinal ailments and neurological symptoms. A single intake of >1-3 mg/kg b.w. is considered a critical effect dose (CED). Probabilistic modelling of acute and chronic (usual) exposure to GAs was performed in the Czech Republic, Sweden and The Netherlands. National databases on individual consumption of foods, data on concentration of GAs in tubers (439 Czech and Swedish results) and processing factors were used for modelling. Results concluded that potatoes currently available at the European market may lead to acute intakes >1 mg GAs/kg b.w./day for upper tail of the intake distribution (0.01% of population) in all three countries. 50 mg GAs/kg raw unpeeled tubers ensures that at least 99.99% of the population does not exceed the CED. Estimated chronic (usual) intake in participating countries was 0.25, 0.29 and 0.56 mg/kg b.w./day (97.5% upper confidence limit). It remains unclear if the incidence of GAs poisoning is underreported or if assumptions are the worst case for extremely sensitive persons. PMID- 19286451 TI - Suppression of nuclear factor kappa B ameliorates astrogliosis but not amyloid burden in APPswe/PS1dE9 mice. AB - Neuroinflammation has been linked to the pathologies of Alzheimer's disease (AD), however, its effects on beta-amyloid (Abeta) burden are unclear. This study investigated the role of nuclear factor kappa B (NF-kappaB) in regulating neuroinflammation and Abeta deposition in a transgenic mouse model of AD. The APPswe/PS1dE9 mice and their wild-type controls received either the NF-kappaB inhibitor pyrrolidine dithiocarbamate (PDTC, i.p. 50 mg/kg daily) or saline starting at 7 months of age for 5 months. Expression of cyclooxygenase-2 (COX-2), tissue necrosis factor alpha (TNFalpha) precursor protein and microtubule associated protein 2 was determined, and astrogliosis was assessed. Hippocampal and cortical levels of Abeta(1-40) and Abeta(1-42) were measured using ELISA. PDTC treatment effectively suppressed NF-kappaB signaling in APPswe/PS1dE9 mice as evidenced by the abolishment of COX-2 and TNFalpha induction. Inhibition of NF kappaB further attenuated astrogliosis in the transgenic AD mice, yet markedly increased cerebral Abeta(1-42) burden. Our findings suggest that NF-kappaB can mediate induction of COX-2, TNFalpha and astrogliosis in APPswe/PS1dE9 mice. Additionally, these results support the idea that neuroinflammation contributes to the clearance of Abeta. PMID- 19286452 TI - Ebselen attenuates cisplatin-induced ROS generation through Nrf2 activation in auditory cells. AB - Ebselen, an organoselenium compound that acts as a glutathione peroxidase mimetic, has been demonstrated to possess antioxidant and anti-inflammatory activities. However, the molecular mechanism underlying this effect is not fully understood in auditory cells. The purpose of the present study is to investigate the protective effect of ebselen against cisplatin-induced toxicity in HEI-OC1 auditory cells, organotypic cultures of cochlear explants from two-day postnatal rats (P(2)) and adult Balb/C mice. Pretreatment with ebselen ameliorated apoptotic death induced by cisplatin in HEI-OC1 cells and organotypic cultures of Corti's organ. Ebselen pretreatment also significantly suppressed cisplatin induced increases in intracellular reactive oxygen species (ROS), intracellular reactive nitrogen species (RNS) and lipid peroxidation levels. Ebselen dose dependently increased the expression level of an antioxidant response element (ARE)-luciferase reporter in HEI-OC1 cells through the translocation of Nrf2 into the nucleus. Furthermore, we found that pretreatment with ebselen significantly restored Nrf2 function, whereas it ameliorated the cytotoxicity of cisplatin in cells transfectants with either a pcDNA3.1 (control) or a DN-Nrf2 (dominant negative) plasmid. We also observed that Nrf2 activation by ebselen increased the expression of phase II antioxidant genes, including heme oxygenase (HO-1), NAD(P)H:quinine oxidoreductase, and gamma-glutamylcysteine synthetase (gamma GCS). Treatment with ebselen resulted in an increased expression of HO-1 and intranuclear Nrf2 in hair cells of organotypic cultured cochlea. After intraperitoneal injection with cisplatin, auditory brainstem responses (ABRs) threshold was measured on 8th day in Balb/C mice. ABR threshold shift was marked occurred in mice injected with cisplatin (16 mg/kg, n=5; Click and 8-kHz stimuli, p<0.05; 4, 16 and 32 kHz, p<0.01), whereas that of animal group which was treated with cisplatin and ebselen was not significantly changed. These results suggest that ebselen activates the Nrf2-ARE signaling pathway, which ultimately prevents free radical stresses from cisplatin and further contributes to protect auditory sensory hair cells from free radicals produced by cisplatin. PMID- 19286453 TI - Nitrite-glucose-glucose oxidase system directly induces rat heart homogenate oxidation and tyrosine nitration: effects of some flavonoids. AB - Protein tyrosine nitration is a common post-translational modification occurring under conditions of nitrative/oxidative stress in a number of diseases. It has been found that in the presence of nitrite and hydrogen peroxide, hemoprotein catalyzes protein tyrosine nitration. In this paper, it was found that in heart homogenate, protein nitration and oxidation could be induced by a nitrite-glucose glucose oxidase system without addition of exogenous heme or hemoprotein. Several structural diversity flavonoids (quercetin, rutin, baicalein, baicalin, apigenin, puerarin, and (+)-catechin) could, more or less, protect rat heart homogenate from oxidative and nitrative injury induced by nitrite-glucose-glucose oxidase in vitro. The inhibitory effects of flavonoids on protein nitration and lipid peroxidation were consistent with their antioxidant activities, whereas the inhibitory effects on protein oxidation were almost contrary to their antioxidant activities. These results mean that nitrite-glucose-glucose oxidase system can cause heart homogenate protein nitration and protein oxidation in different pathways, and those flavonoids with strong antioxidant activities may contribute their protective effect partly through inhibiting protein nitration. PMID- 19286454 TI - The new IS1595 family, its relation to IS1 and the frontier between insertion sequences and transposons. AB - A picture of insertion sequence (IS) diversity is emerging in which previously well-defined groups or families, while remaining tightly clustered, no longer have defined borders but tend to be joined by ISs with partially shared characteristics. This is to some extent due to the presence or absence of different structurally defined transposase domains (and their spacing) and sequence similarities between IS ends. A surprising result arising from the detailed analysis of ISs in various bacterial genomes is the presence of close relatives carrying passenger genes (e.g. antibiotic resistances, methyltransferases, or transcriptional regulators as well as unknown functions). This is beginning to obscure the previously defined line between ISs (no additional orfs) and transposons. We include the lowercase prefix "t" to distinguish them from classical ISs. This is illustrated here by the IS1595 family, distantly related to IS1, which can be subdivided into several groups based on BLAST analysis of transposases, the genetic organization (number and position of the orfs) and the inverted repeats at their ends. The classification was subsequently confirmed using MCL (Markov cluster algorithm) software with parameters derived from the well-defined IS3 family. Many new ISs were identified from the public databases using a reiterative BLAST approach. PMID- 19286455 TI - Biosimilars--science, status, and strategic perspective. AB - Biopharmaceuticals based on recombinant proteins have started to go off-patent, opening the way for other manufacturers to place follow-on products to the market. Meanwhile it has been recognized by all stakeholders that there are fundamental differences between conventional small-molecule based drugs and biopharmaceuticals. This has led to the adoption of distinct legal and regulatory frameworks for biosimilars (follow-on products to biopharmaceuticals) in various parts of the world. This review gives an overview on the scientific basis for the approval requirements, the regulatory and market status, open issues, and the strategic perspective. PMID- 19286456 TI - Hybridization of an ITS-based macroarray with ITS community probes for characterization of complex communities of fungi and fungal-like protists. AB - The ability to characterize fungal community structure and dynamics in the environment is constantly challenged by the high levels of diversity that must be confronted. Large-scale oligonucleotide arrays for use in such analytical studies are currently under development; however, the implementation of this approach generally requires substantial time and financial resources. To address the need for a more accessible tool for fungal community profiling and broad diagnostics, we evaluated the potential utility of a reverse dot blot approach utilizing macroarray targets and probes that each consisted of a PCR product of the entire fungal ITS1-5.8S-ITS2 gene region. Samples used to generate the array targets included both culturable and non-culturable fungi and fungal-like protists representing a range of ecological functions. Tests performed using single species probes within the genus Pythium demonstrated that taxonomic lineages could generally be distinguished when ITS DNA sequence similarity differed by greater than 5-10%. An artificially constructed community probe of known composition successfully detected eight of the 10 lineages contained on the array with only one clear false positive in 95 targets. The approach was also successfully applied to environmental samples. Taxa resident in the soil of a local orchard were identified using the array and matched those documented in previous studies. Closely related taxa from a previously uncharacterized and geographically distant orchard soil were also identified by the array and had affinities to Leptodontium, Cadophora, Zalerion, and Geomyces. These taxa were further confirmed to be present in the sample by cloning and DNA sequencing. A minority of lineages had DNA targets with low melting temperatures which were not detected on the arrays except under conditions that compromised specificity. Membrane-based ITS macroarrays coupled with community ITS probes possessed sufficient power to detect multiple genus-level lineages of fungi in complex samples and should have broad applications in the study of fungal communities. PMID- 19286457 TI - NOV/CCN3 promotes maturation of cerebellar granule neuron precursors. AB - A body of evidence points to the matricial CCN proteins as key regulators of organogenesis. NOV/CCN3, a founder CCN member, is expressed in the developing central nervous system but its functions during neural development have not been studied yet. Here we describe the pattern of NOV expression during rat cerebellar postnatal development and show that NOV expression increases during the second postnatal week, a critical period for the maturation of granule neuron precursors (GNP). NOV transcripts are specifically produced by Purkinje neurons and NOV protein localises extracellularly in the molecular layer and the inner part of the external granule layer, at a key position to control GNP proliferation and migration. In vitro, NOV reduces Sonic Hedgehog-induced GNP proliferation through beta3 integrins and stimulation of GSK3-beta activity whereas NOV stimulates GNP migration through distinct RGD-dependent integrins. These findings identify a new paracrine role of NOV in the development of cerebellar granule neurons. PMID- 19286458 TI - Selection and analysis of HIV-1 integrase strand transfer inhibitor resistant mutant viruses. AB - This report describes methods for the selection and analysis of antiretroviral resistance to HIV integrase strand transfer inhibitors (InSTIs) in cell culture. The method involves the serial passage of HIV-1 in the presence of increasing concentrations of test inhibitors, followed by the cloning and sequencing of the integrase coding region from the selected viruses. The identified mutations are subsequently re-engineered into a reference wild-type molecular clone, and the resulting replication capacity and level of drug resistance are determined relative to the wild-type virus. Here we describe examples of selection and analysis of InSTI-resistant viruses using four integrase inhibitors from three structurally distinct chemical classes; a diketo acid, two naphthyridines, and a pyrimidinecarboxamide. Each inhibitor selected an independent route to resistance. Interestingly, the shift in the IC50 required to suppress the re engineered resistant mutant viruses closely matched the concentration of compound used during the selection of drug resistance. PMID- 19286459 TI - Expression, purification and characterization of Gloydius shedaoensis venom gloshedobin as Hsp70 fusion protein in Pichia pastoris. AB - Gloshedobin, a thrombin-like enzyme from the venom of Gloydius shedaoensis was expressed as Hsp70 fusion protein from the construct pPIC9K/hsp70-TLE in the yeast Pichia pastoris. By fusing gloshedobin to the C-terminus of Hsp70, an expression level of 44.5mg Hsp70-gloshedobin per liter of culture was achieved by methanol induction. The fusion protein secreted in the culture medium was conveniently purified by two chromatographic steps: Q-Sepharose FF and Superdex 200. The purified enzyme had an apparent molecular mass of 98 kDa according to SDS-PAGE analysis, and exhibited fibrinogenolytic activity that preferentially degraded fibrinogen alpha-chain. The enzyme also degraded fibrinogen beta-chain to a lesser extent, while showing no degradation toward the gamma-chain. A fibrinogen clotting activity of 499.8 U/mg was achieved by the enzyme, which is within the range reported for other thrombin-like enzymes. Hsp70-gloshedobin had strong esterase activity toward the chromogenic substrate N alpha-p-tosyl-Gly-Pro Arg-p-nitroanilide, and this activity was optimal at pH 7.5 and 50 degrees C, and was completely inhibited by PMSF, but not by EDTA. We concluded that Hsp70 has no effect on the physiochemical and biochemical properties of gloshedobin. Although applying a fusion partner with very big molecular weight is unusual, Hsp70 proved its advantage in soluble expression of gloshedobin without affecting its fibrinogenolytic activity. And this positive result may provide an alternative strategy for the expression of thrombin-like enzymes in microbial system. PMID- 19286460 TI - A novel framework for segmentation of deep brain structures based on Markov dependence tree. AB - The aim of this work is to develop a new framework for multi-object segmentation of deep brain structures (caudate nucleus, putamen and thalamus) in medical brain images. Deep brain segmentation is difficult and challenging because the structures of interest are of relatively small size and have significant shape variations. The structure boundaries may be blurry or even missing, and the surrounding background is full of irrelevant edges. To tackle these problems, we propose a template-based framework to fuse the information of edge features, region statistics and inter-structure constraints for detecting and locating all target brain structures such that initialization by hand is unnecessary. The multi-object template is organized in the form of a hierarchical Markov dependence tree (MDT), and multiple objects are efficiently matched to a target image by a top-to-down optimization strategy. The final segmentation is obtained through refinement by a B-spline based non-rigid registration between the exemplar image and the target image. Our approach needs only one example as training data. We have validated the proposed method on a publicly available T1 weighted magnetic resonance image database with expert-segmented brain structures. In the experiments, the proposed approach has obtained encouraging results with 0.80 Dice score for the caudate nuclei, 0.81 Dice score for the putamina and 0.84 Dice score for the thalami on average. PMID- 19286461 TI - Magnetic resonance microscopy of mammalian neurons. AB - Magnetic resonance imaging (MRI) is now a leading diagnostic technique. As technology has improved, so has the spatial resolution achievable. In 1986 MR microscopy (MRM) was demonstrated with resolutions in the tens of micrometers, and is now an established subset of MRI with broad utility in biological and non biological applications. To date, only large cells from plants or aquatic animals have been imaged with MRM limiting its applicability. Using newly developed microsurface coils and an improved slice preparation technique for correlative histology, we report here for the first time direct visualization of single neurons in the mammalian central nervous system (CNS) using native MR signal at a resolution of 4-8 microm. Thus MRM has matured into a viable complementary cellular imaging technique in mammalian tissues. PMID- 19286462 TI - Quantitative analysis of donepezil binding to acetylcholinesterase using positron emission tomography and [5-(11)C-methoxy]donepezil. AB - The aim of this study was to establish kinetic analysis of [5-(11)C methoxy]donepezil ([(11)C]donepezil), which was developed for the in-vivo visualization of donepezil binding to acetylcholinesterase (AChE) using positron emission tomography (PET). Donepezil is an AChE inhibitor that is widely prescribed to ameliorate the cognitive impairment of patients with dementia. Six healthy subjects took part in a dynamic study involving a 60-min PET scan after intravenous injection of [(11)C]donepezil. The total distribution volume (tDV) of [(11)C]donepezil was quantified by compartmental kinetic analysis and Logan graphical analysis. A one-tissue compartment model (1TCM) and a two-tissue compartment model (2TCM) were applied in the kinetic analysis. Goodness of fit was assessed with chi(2) criterion and Akaike's Information Criterion (AIC). Compared with a 1TCM, goodness of fit was significantly improved by a 2TCM. The tDVs provided by Logan graphical analysis were slightly lower than those provided by a 2TCM. The rank order of the mean tDVs in 10 regions was in line with the AChE activity reported in a previous post-mortem study. Logan graphical analysis generated voxel-wise images of tDV, revealing the overall distribution pattern of AChE in individual brains. Significant correlation was observed between tDVs calculated with and without metabolite correction for plasma time-activity curves, indicating that metabolite correction could be omitted. In conclusion, this method enables quantitative analysis of AChE and direct investigation of the pharmacokinetics of donepezil in the human brain. PMID- 19286463 TI - Study of neurovascular coupling in humans via simultaneous magnetoencephalography and diffuse optical imaging acquisition. AB - By combining diffuse optical imaging (DOI) and magnetoencephalography (MEG) we investigate neurovascular coupling non-invasively in human subjects using median nerve stimulation. Previous fMRI studies have shown a habituation effect in the hemodynamic blood oxygen level-dependent (BOLD) response for stimulation periods longer than 2 s. With DOI and MEG we can test whether this effect in hemodynamic response can be accounted for by a habituation effect in the neural response. Our experimental results show that the habituation effect in the hemodynamic response is stronger than that in the earliest cortical neural response (N20). Using a linear convolution model to predict hemodynamic responses we found that including late neural components (> or = 30 ms) improves the prediction of the hemoglobin response. This finding suggests that in addition to the initial evoked-response deflections related to the talamic afferent input, later cortical activity is needed to predict the hemodynamic response. PMID- 19286464 TI - Connectivity alterations assessed by combining fMRI and MR-compatible hand robots in chronic stroke. AB - The aim of this study was to investigate functional reorganization of motor systems by probing connectivity between motor related areas in chronic stroke patients using functional magnetic resonance imaging (fMRI) in conjunction with a novel MR-compatible hand-induced, robotic device (MR_CHIROD). We evaluated data sets obtained from healthy volunteers and right-hand-dominant patients with first ever left-sided stroke > or =6 months prior and mild to moderate hemiparesis affecting the right hand. We acquired T1-weighted echo planar and fluid attenuation inversion recovery MR images and multi-level fMRI data using parallel imaging by means of the GeneRalized Autocalibrating Partially Parallel Acquisitions (GRAPPA) algorithm on a 3 T MR system. Participants underwent fMRI while performing a motor task with the MR_CHIROD in the MR scanner. Changes in effective connectivity among a network of primary motor cortex (M1), supplementary motor area (SMA) and cerebellum (Ce) were assessed using dynamic causal modeling. Relative to healthy controls, stroke patients exhibited decreased intrinsic neural coupling between M1 and Ce, which was consistent with a dysfunctional M1 to Ce connection. Stroke patients also showed increased SMA to M1 and SMA to cerebellum coupling, suggesting that changes in SMA and Ce connectivity may occur to compensate for a dysfunctional M1. The results demonstrate for the first time that connectivity alterations between motor areas may help counterbalance a functionally abnormal M1 in chronic stroke patients. Assessing changes in connectivity by means of fMRI and MR_CHIROD might be used in the future to further elucidate the neural network plasticity that underlies functional recovery in chronic stroke patients. PMID- 19286465 TI - Spatial and temporal reproducibility-based ranking of the independent components of BOLD fMRI data. AB - Independent component analysis (ICA) decomposes fMRI data into spatially independent maps and their corresponding time courses. However, distinguishing the neurobiologically and biophysically reasonable components from those representing noise and artifacts is not trivial. We present a simple method for the ranking of independent components, by assessing the resemblance between components estimated from all the data, and components estimated from only the odd- (or even-) numbered time points. We show that the meaningful independent components of fMRI data resemble independent components estimated from downsampled data, and thus tend to be highly ranked by the method. PMID- 19286466 TI - IGF-1 and G-CSF complement each other in BMSC migration towards infarcted myocardium in a novel in vitro model. AB - Stem cell capability enhanced with cytokine administration is a promising treatment for myocardial infarction. Bone marrow stem cells (BMSCs) were isolated from C57BL/6 mice (8-12 weeks old) expressing GFP and characterized with c-kit and CD34. Infarcted heart tissue fragments were placed into dishes with BMSCs and medium supplemented with G-CSF, SCF, IGF-1 or combinations thereof were given to the BMSC-infarcted myocardium in vitro model. The IGF-1-G-CSF group showed significantly higher migration (67.7% +/- 2.6) of c-kit(+) BMSCs towards the ischemic tissue and expressed MEF-2 (43.7% +/- 1.7). Of the single treatment groups, the G-CSF group demonstrated significantly higher migration of c-kit(+) BMSCs (60.5 +/- 2.7) with MEF-2 expression (38.7 +/- 1.4). IGF-1 complements G CSF and was relatively more significant in its effects on BMSC migration and cardiac lineage commitment towards ischemic heart tissue. PMID- 19286467 TI - Rat marrow-derived mesenchymal stem cells developed in a medium supplemented with the autologous versus bovine serum. AB - The controversial effect of autologous serum (AS) on human mesenchymal stem cells (MSC) was studied in rat MSC culture. Rat bone marrow cells were plated in a medium containing either FBS (fetal bovine serum) or AS were cultured to passage 3, during which the population doubling number (PDN) of both cultures were measured and compared statistically. The number of viable cells, the cell colonogic activity, and cell growth rate were also compared. In addition, mineralization in the osteogenic cultures from each system was measured. Our data indicated that AS enriched medium provided a microenvironment in which growth rate as well as bone differentiation of the isolated MSCs were significantly higher than in FBS enriched medium. PMID- 19286468 TI - Pre-adipocytes commitment to neurogenesis 1: preliminary localisation of cholinergic molecules. AB - A great effort has recently been made to obtain human stem cells able to differentiate into cholinergic neurons, as a number of diseases are associated to the cholinergic neuron loss, degeneration or incorrect function (Alzheimer's disease and motor neuron disease). A stem cell population (i.e. pre-adipocytes) is present in the adipose stromal compartment. Pre-adipocytes, like the mesodermic derivative cells, retain high plasticity and potentiality to convert in vitro from one phenotype into many others, and they can be isolated from adult adipose tissue. Pre-adipocytes committed in vitro to neural differentiation were followed up to the acquisition of neural morphology. Acetylcholinesterase and choline acetyltransferase are expressed from the native cell stage, with different localisations and roles during neural commitment. Western blots show the beginning of a new synthesis of these enzymes at 4 weeks of culture of neurogenic pre-adipocytes, in parallel with neural morphology. The passage of the choline-acetyltransferase immunoreactivity from cytoplasmic to membrane localisation shows the possible onset of catalytic activity and the histochemical reaction confirms the activity of acetylcholinesterase. This explains the possibility of obtaining cholinergic-like phenotype from pre-adipocytes. PMID- 19286469 TI - ST2 deficient mice display a normal host defense against pulmonary infection with Mycobacterium tuberculosis. AB - Tuberculosis, caused by Mycobacterium (M.) tuberculosis, is a devastating infectious disease causing many deaths world-wide every year. Successful host defense mainly depends on a strong Th type 1 response. We investigated the role of T1/ST2 (recently identified as the receptor for IL-33), a typical Th2 marker in the assumption that a shift towards a beneficial Th1 response would occur in the absence of ST2. For this, ST2 KO and WT mice were intranasally infected with a virulent strain of M. tuberculosis (150 CFU). In line with our hypothesis, ST2 KO animals displayed increased numbers of lymphocytes infiltrating the lung after 2 weeks of infection, increased IFNgamma production by splenocytes in ST2 KO mice early in infection and enhanced lung IFNgamma levels at the chronic phase of the disease. However, we did not detect any differences between ST2 KO and WT mice in mycobacterial loads in lungs or liver after M. tuberculosis infection. The pulmonary inflammatory response, as measured by relative lung weights, cytokine and chemokine levels as well as histopathological analysis, was similar in ST2 KO and WT mice. These data suggest that apart from inducing a modest shift towards the Th1 response, the role of ST2 during murine M. tuberculosis infection is limited. PMID- 19286470 TI - Linking phospholipid flippases to vesicle-mediated protein transport. AB - Type IV P-type ATPases (P4-ATPases) are a large family of putative phospholipid translocases (flippases) implicated in the generation of phospholipid asymmetry in biological membranes. P4-ATPases are typically the largest P-type ATPase subgroup found in eukaryotic cells, with five members in Saccharomyces cerevisiae, six members in Caenorhabditis elegans, 12 members in Arabidopsis thaliana and 14 members in humans. In addition, many of the P4-ATPases require interaction with a noncatalytic subunit from the CDC50 gene family for their transport out of the endoplasmic reticulum (ER). Deficiency of a P4-ATPase (Atp8b1) causes liver disease in humans, and studies in a variety of model systems indicate that P4-ATPases play diverse and essential roles in membrane biogenesis. In addition to their proposed role in establishing and maintaining plasma membrane asymmetry, P4-ATPases are linked to vesicle-mediated protein transport in the exocytic and endocytic pathways. Recent studies have also suggested a role for P4-ATPases in the nonvesicular intracellular trafficking of sterols. Here, we discuss the physiological requirements for yeast P4-ATPases in phospholipid translocase activity, transport vesicle budding and ergosterol metabolism, with an emphasis on Drs2p and its noncatalytic subunit, Cdc50p. PMID- 19286472 TI - Modulation of phospholipase D activity in vitro. AB - Most phospholipases D (PLDs) occurring in microorganisms, plants and animals belong to a superfamily which is characterized by several conserved regions of amino acid sequence including the two HKD motifs necessary for catalytic activity. Most eukaryotic PLDs possess additional regulatory structures such as the Phox and Pleckstrin homology domains in mammalian PLDs and the C2 domain in most plant PLDs. Owing to recombinant expression techniques, an increasing number of PLDs from different organisms has been obtained in purified form, allowing the investigation of specific and unspecific interactions of the enzymes with regulatory components in vitro. The present paper gives an overview on different factors which can modulate PLD activity and compares their influence on the enzymes from different sources. While no biological regulator can be recognized for extracellular bacterial PLDs, the most prominent specific activator of eukaryotic PLDs is phosphatidylinositol-4,5-bisphosphate (PIP(2)). In a sophisticated interplay PIP(2) seems to cooperate with several regulatory proteins in mammalian PLDs, whereas in plant PLDs it mainly acts in concert with Ca(2+) ions. Moreover, curvature, charges and heterogeneities of membrane surfaces are assessed as unspecific modulators. A possible physiological role of the transphosphatidylation reaction catalyzed by PLDs in competition with phospholipid hydrolysis is discussed. PMID- 19286471 TI - Intracellular sterol dynamics. AB - We review the cellular mechanisms implicated in cholesterol trafficking and distribution. Recent studies have provided new information about the distribution of sterols within cells, including analysis of its transbilayer distribution. The cholesterol interaction with other lipids and its engagement in various trafficking processes will determine its proper level in a specific membrane; making the cholesterol distribution uneven among the various intracellular organelles. The cholesterol content is important since cholesterol plays an essential role in membranes by controlling their physicochemical properties as well as key cellular events such as signal transduction and protein trafficking. Cholesterol movement between cellular organelles is highly dynamic, and can be achieved by vesicular and non-vesicular processes. Various studies have analyzed the proteins that play a significant role in these processes, giving us new information about the relative importance of these two trafficking pathways in cholesterol transport. Although still poorly characterized in many trafficking routes, several potential sterol transport proteins have been described in detail; as a result, molecular mechanisms for sterol transport among membranes start to be appreciated. PMID- 19286474 TI - Gelastic seizures: A case of lateral frontal lobe epilepsy and review of the literature. AB - We describe a 40-year-old patient with gelastic seizures triggered by hand movement. Despite nonlesional magnetic resonance imaging (MRI), electroencephalography (EEG), functional magnetic resonance imaging (fMRI), and diffusion tensor imaging (DTI) are concordant with seizure onset in the right frontocentral area. Seizure semiology and EEG recordings imply involvement of mesial frontal structures remote from seizure initiation site. We reviewed all published cases on gelastic seizures of frontal lobe origin to find characteristic features. For further investigation of the phenomenon of movement induced seizures, fMRI was performed using a finger tapping paradigm. Interictal fMRI revealed widespread activation of right motor cortex during finger tapping on either side outreaching the anatomical representation of the left finger. In line with this finding DTI revealed fiber track impairment in the right frontocentral region, supporting the hypothesis of a focal derangement. This case highlights the importance of complementary functional investigations in MRI negative epilepsies. PMID- 19286475 TI - Quantitative MRI of the prefrontal cortex and executive function in patients with temporal lobe epilepsy. AB - We investigated the relationship between prefrontal cortex (PFC) and hippocampal volume and executive functioning in patients with temporal lobe epilepsy (TLE). Prefrontal volume and hippocampal volume were studied using stereology in conjunction with point counting and voxel-based morphometry on MR images. Executive functioning was assessed using tests routinely incorporated into presurgical neuropsychological evaluation. Relative to 30 healthy controls, 43 patients (26 left, 17 right) with TLE had volume atrophy of the ipsilateral hippocampus and bilateral dorsal PFC. Performance on the working memory index of the Wechsler Memory Scale was positively correlated with the volume of all prefrontal regions, and the Controlled Oral Word Association Test with the left dorsal PFC, whole left PFC, and left hippocampus. Stroop Color-Word Interference performance was not related to volume of dorsal PFC. The "extratemporal neuropsychological profile" frequently observed in patients with TLE may be due to extended damage to brain regions remote from the epileptogenic focus. In particular, volume atrophy of the dorsal PFC may account for deficits in executive functioning. PMID- 19286473 TI - Cholesterol transport in steroid biosynthesis: role of protein-protein interactions and implications in disease states. AB - The transfer of cholesterol from the outer to the inner mitochondrial membrane is the rate-limiting step in hormone-induced steroid formation. To ensure that this step is achieved efficiently, free cholesterol must accumulate in excess at the outer mitochondrial membrane and then be transferred to the inner membrane. This is accomplished through a series of steps that involve various intracellular organelles, including lysosomes and lipid droplets, and proteins such as the translocator protein (18 kDa, TSPO) and steroidogenic acute regulatory (StAR) proteins. TSPO, previously known as the peripheral-type benzodiazepine receptor, is a high-affinity drug- and cholesterol-binding mitochondrial protein. StAR is a hormone-induced mitochondria-targeted protein that has been shown to initiate cholesterol transfer into mitochondria. Through the assistance of proteins such as the cAMP-dependent protein kinase regulatory subunit Ialpha (PKA-RIalpha) and the PKA-RIalpha- and TSPO-associated acyl-coenzyme A binding domain containing 3 (ACBD3) protein, PAP7, cholesterol is transferred to and docked at the outer mitochondrial membrane. The TSPO-dependent import of StAR into mitochondria, and the association of TSPO with the outer/inner mitochondrial membrane contact sites, drives the intramitochondrial cholesterol transfer and subsequent steroid formation. The focus of this review is on (i) the intracellular pathways and protein-protein interactions involved in cholesterol transport and steroid biosynthesis and (ii) the roles and interactions of these proteins in endocrine pathologies and neurological diseases where steroid synthesis plays a critical role. PMID- 19286476 TI - Health, wealth, and culture as predominant factors in psychosocial morbidity. AB - Depression is the most common psychological morbidity in epilepsy, yet this comorbidity is not well understood. Possible explanations for this comorbidity include recurrence of premorbid depression, increased risk for severe epilepsy due to a history of depression, shared risk factors for depression and epilepsy, AED-induced depression in vulnerable individuals, and coping styles in the face of stressors linked to epilepsy. Preexisting vulnerability to depression may contribute to each of these explanations. Vulnerability may arise from the influence of common risk factors, family history of depression, a history of depression before initiation of relevant AEDs, or coping styles and may reflect allostatic load. These exposures may precede the occurrence of epilepsy or follow the onset of epilepsy, in both cases increasing the risk for depression in prevalent epilepsy. Their careful evaluation is vital to identifying people at greatest risk for depression in epilepsy and for informing interventions to prevent the occurrence of this disabling epilepsy comorbidity. PMID- 19286477 TI - Primary antral duodenal extramedullary plasmacytoma presenting with melena. PMID- 19286478 TI - What makes individuals with gastroesophageal reflux disease dissatisfied with their treatment? AB - BACKGROUND & AIMS: Despite the efficacy of proton pump inhibitors (PPIs) as therapeutics for patients with gastroesophageal reflux disease (GERD) in randomized controlled trials, a number of studies have shown that a proportion of patients with GERD are not satisfied with their treatment. This article reviews the possible reasons why patients are dissatisfied with the way their disease is managed. METHODS: Studies published between 1970 and 2007 were identified from PubMed, EMBASE, and the author's existing database. The 2708 publications were reviewed, and irrelevant ones were excluded. Eleven studies were found to be appropriate for use in this review. RESULTS: Patients who are given prescriptions for PPIs tend to be more satisfied than those given H(2)-receptor antagonists. Partial responders are likely to be more dissatisfied than patients whose symptoms are fully resolved. A decrease in health-related quality of life is associated with greater dissatisfaction. Patients are more likely to be satisfied if they are taken seriously by their physician and if their symptoms are investigated. They are also more likely to be satisfied if the patient-physician consultation is interactive. CONCLUSIONS: Patient satisfaction is a complex issue that depends on many factors. Patient satisfaction can be influenced by treatment regimen, general level of well-being, the bedside manner of the physician, and the quality of patient-physician communication. Improvements in recognition of GERD can improve management of the disease as well as patient satisfaction with their care and treatment. PMID- 19286479 TI - Gardner variant of familial adenomatous polyposis: from extensive skull osteomatosis to metastatic rectal remnant cancer. PMID- 19286480 TI - Face to face with teniasis. PMID- 19286481 TI - A population-based study showing an association between gastroesophageal reflux disease and sleep problems. AB - BACKGROUND & AIMS: Gastroesophageal reflux disease and sleep problems are common health problems in Western nations. It is important to clarify the association between sleep and gastroesophageal reflux disease, but only a few population based studies have been conducted. METHODS: A population-based, cross-sectional, case-control study was based on 2 large health surveys performed in the Norwegian county Nord-Trondelag in 1984-1986 and 1995-1997. Gastroesophageal reflux disease was assessed in the second survey, which included 65,333 participants (70% of the county's adult population). The 3153 persons who reported severe reflux symptoms constituted the cases, and the 40,210 persons without reflux symptoms constituted the controls. Data on insomnia, sleep problems, and several potential confounders were collected in questionnaires. Odds ratios (ORs) with 95% confidence intervals (CIs) were estimated by using unconditional logistic regression in crude and multivariable models. RESULTS: In models adjusted for age, sex, tobacco smoking, obesity, and socioeconomic status, positive associations were observed between presence of insomnia (OR, 3.2; 95% CI, 2.7-3.7), sleeplessness (OR, 3.3; 95% CI, 2.9-3.8), problems falling asleep (OR, 3.1; 95% CI, 2.5-3.8), and risk of gastroesophageal reflux disease. These associations were attenuated after further adjustments for anxiety, depression, myocardial infarction, angina pectoris, stroke, and gastrointestinal symptoms, but they remained statistically significant. CONCLUSIONS: A large population-based study indicated a link between sleep problems and gastroesophageal reflux disease that might be bidirectional. PMID- 19286482 TI - Dietary supplementation with chitosan derived from mushrooms changes adipocytokine profile in diet-induced obese mice, a phenomenon linked to its lipid-lowering action. AB - Recent data reported that chitosan reduces high-fat (HF) diet-induced obesity in mice without describing the metabolic consequences of such an effect. The aim of this study was to investigate the capacity of chitosan derived from edible mushrooms to modify adipocytokine levels and to assess the relevance of this effect on the development of fat mass, and on glucose and lipid metabolism in obese mice. Mice were fed a HF diet or a HF diet supplemented with 5% fungal chitosan for ten weeks. HF-induced hypertriglyceridaemia, fasting hyperinsulinaemia and fat accumulation in liver, muscle and white adipose tissue (WAT) were reduced after chitosan treatment. The higher lipid content in the caecum following treatment with chitosan suggested that this dietary fiber reduced lipid absorption. We postulated that the lower triglyceridaemia observed upon chitosan treatment could also be the result of the lower FIAF (fasting induced adipose factor) expression observed in visceral adipose tissue. IL-6, resistin and leptin levels decreased in the serum after chitosan supplementation. We conclude that fungal chitosan counteracts some inflammatory disorders and metabolic alterations occurring in diet-induced obese mice since it decreases feed efficiency, fat mass, adipocytokine secretion and ectopic fat deposition in the liver and the muscle. PMID- 19286483 TI - Decreased inflammation and augmented expression of trophic factors correlate with MOG-induced neuroprotection of the injured nigrostriatal system in the murine MPTP model of Parkinson's disease. AB - The response of the immune system during injury of the central nervous system may play a role in protecting neurons. We have previously reported that immunization with MOG 35-55 prior to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced injury of the dopaminergic system promotes less dopamine depletion and less dopaminergic damage of neurons in mice. In this study, we evaluate the influence of MOG immunization on the inflammatory reaction that occurs at the place of injury. C57Bl male mice, 2 and 12 months old, received i.p. injections of MPTP (40 mg/kg) and some groups animals also received an additional injection with myelin oligodendrocyte glycoprotein (MOG) 35-55 in CFA 6 days before MPTP administration. MPTP caused a common inflammatory reaction characterized by microglial activation, infiltration of T cells into the substantia nigra and striatum and increased expression of mRNA encoding pro-inflammatory cytokines (IL 1 beta, TNFalpha, INF gamma) and trophic factors (TGFbeta, GDNF). MOG immunization prior to MPTP administration significantly diminished the microglial reaction and reduced the levels of infiltrating CD8+ lymphocytes. The number of CD4+ T cells remained at the same level as in the MPTP group. Expression of pro inflammatory cytokines was diminished. The mRNA expression of GDNF was significantly higher in the MOG pretreated mice relative to the MPTP group, both in the 2 month old and 12 month old groups. Since MOG immunization prior to MPTP intoxication appears to prevent nigrostriatal injury, the observed decrease of inflammation and increase of GDNF mRNA expression in the injured areas might represent one of the mechanisms of observed neuroprotection. PMID- 19286484 TI - Differential stabilities of alternative exon-skipped rod motifs of dystrophin. AB - Exon skipping repair is a strategy being investigated in early stage clinical trials to treat Duchenne muscular dystrophy. This is most applicable to the majority of cases which arise when genetic defects cause frame shift mutations, and induced exon skipping of out-of-phase exons restores the reading frame. However, the consequences to the edited protein so produced have not been considered. In many cases alternative routes to restoring the reading frame are possible, and we show in a test case involving exon 44 that the resulting differently edited proteins greatly vary in stability, with one of them very similar to normal unskipped dystrophin, and the other much less stable as assessed by the thermodynamics of folding as well as resistance to proteolysis. This has implications for the design of optimal therapeutic exon skipping strategies, which presumably wish to result repairs with as much fidelity to normal dystrophin as possible. PMID- 19286485 TI - Severe proctitis due to lymphogranuloma venereum (Chlamydia trachomatis). PMID- 19286486 TI - Syphilis surveillance in Portugal, 2000-2005. PMID- 19286487 TI - Leishmaniasis due to L. infantum presenting as macrocheilitis and responding to liposomal amphotericin B. PMID- 19286488 TI - High prevalence of Staphylococcus aureus cultivation and superantigen production in patients with psoriasis. AB - The aim of this study was to evaluate the association of Staphylococcal enterotoxins (se) a through e, exfoliative toxin (et) a and b, toxin and toxic shock syndrome toxin (tst) and mecA with psoriasis. We also investigated the distribution of Staphylococcus aureus (S. aureus) in the skin and nares. Fifty consecutive patients with chronic plaque-type psoriasis and 50 sex- and age matched healthy controls were included in this study. There was a statistical difference in cultivation of S. aureus between lesional (64%) and non-lesional skin (14%) in patients with psoriasis (p = 0.037). S. aureus was cultivated from the nares in 25 (50%) of 50 patients with psoriasis and in 17 (34%) of 50 healthy controls (p > 0.05). In psoriasis patients, 31 (96.8%) out of the 32 strains isolated from the lesional skin and 3 (42.3%) out of the 7 strains isolated from the non-lesional skin were toxigenic (p = 0.01). Isolated strains from the nares were toxigenic in 96% (24/25) for patients with psoriasis and in 41.2% (7/17) for healthy controls, respectively (p = 0.006). Patients with cultivation-positive in lesional skin had a significantly higher PASI score than patients who were cultivation-negative in lesional skin (8.28 +/- 3.97 vs. 5.89 +/- 2.98, p = 0.031). Our results confirm that S. aureus colonization and its toxigenic-strains are associated with psoriasis. According to our findings, non-classical superantigens such as methicillin resistance gene (mecA), see and etb may also be associated with psoriasis. PMID- 19286489 TI - Tongue hyperpigmentation during interferon-alpha and ribavirin therapy. PMID- 19286490 TI - Kinesigenic reflex epilepsy associated with a glioma in the lateral peri-rolandic region. AB - A patient with kinesigenic focal motor seizures induced by tongue-jaw movement had a grade III astrocytoma clearly co-localizing with the epileptic network in the appropriate peri-rolandic, motor-sensory, lingual-jaw cortical area. The clinical seizure phenomena were time-locked with the EEG epileptic activity. [Published with video sequences]. PMID- 19286491 TI - Epileptic negative drop attacks in atypical benign partial epilepsy: a neurophysiological study. AB - PURPOSE: We conducted a computer-assisted polygraphic analysis of drop attacks in a child with atypical benign partial epilepsy (ABPE) to investigate neurophysiological characteristics. SUBJECT AND METHODS: The patient was a six year two-month-old girl, who had started to have focal motor seizures, later combined with daily epileptic negative myoclonus (ENM) and drop attacks, causing multiple injuries. We studied episodes of ENM and drop attacks using video polygraphic and computer-assisted back-averaging analysis. RESULTS: A total of 12 ENM episodes, seven involving the left arm (ENMlt) and five involving both arms (ENMbil), and five drop attacks were captured for analysis. All episodes were time-locked to spike-and-wave complexes (SWC) arising from both centro-temporo parietal (CTP) areas. The latency between the onset of SWC and ENMlt, ENMbil, and drop attacks reached 68 ms, 42 ms, and 8 ms, respectively. The height of the spike as well as the slow-wave component of SWC for drop attacks were significantly larger than that for both ENMlt and ENMbil (p < 0.05). CONCLUSION: Drop attacks were considered to be epileptic negative myoclonus involving not only upper proximal but also axial muscles, causing the body to fall. Thus, drop attacks in ABPE are considered to be epileptic negative drop attacks arising from bilateral CTP foci and differ from drop attacks of a generalized origin seen in Lennox-Gastaut syndrome and myoclonic-astatic epilepsy. PMID- 19286492 TI - Topical imiquimod associated to a reduction of heel hyperkeratosis for the treatment of recalcitrant mosaic plantar warts. PMID- 19286493 TI - Altered EEG spectral activity and attentional behavioral deficits following prolonged febrile seizures: a pilot study. AB - The consequences of febrile seizures (FSs) in infants are still a matter of debate. It is important to develop non-invasive tools to determine markers of brain function that could have predictive value for the outcome of FSs infants.Pattern visual evoked potentials (pVEPs) were recorded in 18 FS infants (mean age of seizure 15.97 months). Spectral density and coherence analyses were performed in infants evaluated at 1.1 month (n = 4), 5.75 months (n = 4) or 30.33 months (n = 6) following a prolonged FS and compared to age-matched healthy controls. The impact of severity of seizures was assessed by comparing the children who had prolonged FSs to 4 infants that had experienced a simple FS. Cognitive tests (Bayley, Stanford-Binet) were administered at the time of testing in FS and control children. Behavioural measures (Achenbach Child Behavior Check List) were administered two years after the FS. pVEP responses and coherence measures failed to yield significant differences between the FS groups and healthy controls. However, spectral density measures showed a significant increase in delta band activity in both FS groups and a reduced high frequency density only in the prolonged FS groups that was seen up to 39 months post seizure. Behavioural and cognitive measures revealed cognitive development within average, but lower attentional capacities in the FS infants. The persistent changes in spectral density patterns seen in children with prolonged FS may reflect seizure induced alterations in the developing brain or a result of a complex mode of inheritance. Further studies are needed to determine whether these observations can be used as a marker to predict the vulnerability of the child in developing behavioral deficits or epilepsy. PMID- 19286494 TI - Quality of life after vagal nerve stimulator insertion. AB - AIM: Assess quality-of-life after vagal nerve stimulation and determine patient characteristics associated with improvement in quality-of-life. METHODS: Sixteen patients (11 children, 5 adults) who had vagal nerve stimulation at our center were studied. Quality-of-life was assessed pre- and post-vagal nerve stimulation using the Quality-of-Life in Childhood Epilepsy questionnaire for children and the Epilepsy Surgery Inventory-55 for adults. RESULTS: Sixteen patients who did not qualify for resective surgery were included; seven (43.75%) were males and 9 (56.25%) were females. Mean age at onset of seizures was 3.96 +/- 4.00 years and at surgery was 15.78 +/- 10.78. Follow-up time was 1.26 +/- 0.92 years. Fourteen patients (87.5%) were mentally retarded. Ten (62.5%) had cryptogenic etiology and 6 patients (37.5%) symptomatic etiology. Fifty percent had localization-related epilepsy. Six of 7 patients with generalized cryptogenic etiology (85.71%) had Lennox-Gastaut syndrome. Seizures dropped from 122.31 +/- 159.49 to 67.84 +/- 88.22 seizures/month. Seizure reduction (> 50%) correlated with improvement in total quality-of-life (p = 0.034). Post-vagal nerve stimulation, the total group scored significantly higher in the social domain (p = 0.039). In patients with localization-related epilepsy, significant improvements were detected in the social domain (p = 0.049) and in total quality-of-life (p = 0.042). CONCLUSION: Despite a diverse and small population size, we observed significant improvements in the social domain 1.26 years post-vagal nerve stimulation. In addition, there was an improvement in total quality-of-life amongst patients with partial seizures. Finally, seizure reduction was associated with quality-of-life improvement. Our results support previous studies from the West reporting improvement in quality-of-life following vagal nerve stimulation, contradict those studies that did not show such differences, and are the first coming from a developing country. PMID- 19286495 TI - Field evaluation of a multiplex real-time reverse transcription polymerase chain reaction assay for detection of Vesicular stomatitis virus. AB - Sporadic outbreaks of vesicular stomatitis (VS) in the United States result in significant economic losses for the U.S. livestock industries because VS is a reportable disease that clinically mimics foot-and-mouth disease. Rapid and accurate differentiation of these 2 diseases is critical because their consequences and control strategies differ radically. The objective of the current study was to field validate a 1-tube multiplexed real-time reverse transcription polymerase chain reaction (real-time RT-PCR) assay for the rapid detection of Vesicular stomatitis New Jersey virus and Vesicular stomatitis Indiana virus strains occurring in Mexico and North and Central America. A comprehensive collection of 622 vesicular lesion samples obtained from cattle, horses, and swine from throughout Mexico and Central America was tested by the real-time RT-PCR assay and virus isolation. Overall, clinical sensitivity and specificity of the real-time RT-PCR were 83% and 99%, respectively. Interestingly, VS virus isolates originating from a specific region of Costa Rica were not detected by real-time RT-PCR. Sequence comparisons of these viruses with the real-time RT-PCR probe and primers showed mismatches in the probe and forward and reverse primer regions. Additional lineage-specific primers and a probe corrected the lack of detection of the missing genetic lineage. Thus, this assay reliably identified existing Mexican and Central American VS viruses and proved readily adaptable as new VS viruses were encountered. An important secondary result of this research was the collection of hundreds of new VS virus isolates that provide a foundation from which many additional studies can arise. PMID- 19286496 TI - Enzyme-linked immunospot: an alternative method for the detection of interferon gamma in Johne's disease. AB - To date, the sensitivity of the interferon gamma (IFN-gamma) enzyme-linked immunosorbent assay (ELISA) to detect Johne's disease (JD) has been poor, especially in the early stages of disease. To improve the sensitivity of IFN gamma detection in the early stages of infection, an alternate assay needs to be developed. The enzyme-linked immunospot (ELISPOT) assay is a highly sensitive technique for the detection of cytokines and has the potential to improve the diagnosis of JD. Of the variables examined, choice of capture antibody and the method by which the peripheral blood mononuclear cells were isolated significantly affected the ability to enumerate IFN-gamma-secreting cells. The ELISPOT assay was as sensitive as or better than the IFN-gamma ELISA at detecting ovine JD and could also detect disease at early time points postinoculation. The IFN-gamma ELISPOT could distinguish infected from unexposed animals; however, neither the IFN-gamma ELISA nor the ELISPOT assay could distinguish between sheep experimentally infected with Mycobacterium avium subspecies paratuberculosis and those exposed to the bacterium but diagnosed as uninfected at necropsy. PMID- 19286497 TI - Immunophenotyping and gene rearrangement analysis in dogs with lymphoproliferative disorders characterized by small-cell lymphocytosis. AB - Lymphocytosis caused by neoplastic proliferation of small lymphocytes is occasionally difficult to distinguish by morphological examination from nonneoplastic lymphocytosis. To examine the clinical utility of gene rearrangement analysis for demonstrating neoplastic proliferation of small lymphocytes, gene rearrangement analysis was performed in comparison with immunophenotyping using peripheral lymphocytes in dogs with small lymphocytosis. Thirty-one dogs with small-cell lymphocytosis (8,100-884,300/microl) were enrolled. By immunophenotyping, lymphocytosis of all dogs was suggested to be neoplastic in nature based on the detection of marked expansion of phenotypically homogeneous lymphocytes or the presence of an aberrant antigen-expressing population of lymphocytes. In contrast, gene rearrangement analysis represented clonality in 27 dogs (detection rate of 87%). From the present study, gene rearrangement analysis was considered to be worthwhile to strengthen the evidence of neoplastic proliferation of small lymphocytes when coupled with immunophenotyping and to be a suitable diagnostic substitute if immunophenotyping is not available in clinical practice. PMID- 19286498 TI - Chromatographic analysis of lipid fractions in healthy dogs and dogs with obesity or hyperadrenocorticism. AB - Obesity and endogenous hyperadrenocorticism (HAC) are common clinical conditions in veterinary practice, and both conditions have clinical and laboratory similarities, such as weight gain and dyslipidemia. The objective of the present study was to characterize and compare the lipid profiles and plasma lipoprotein fractions in healthy dogs (n = 10), in obese dogs (n = 10), and in dogs with HAC (n = 6). All of the dogs were client owned. The lipoproteins were separated by fast protein liquid chromatography, and the plasma concentrations of total cholesterol and total triacylglycerol (TAG) were determined by enzymatic methods. When compared with the healthy and obese groups, dogs with HAC had a significant increase (P < 0.01) in the total concentrations of TAGs and cholesterol (CHOL), with higher distribution in the very low-density lipoprotein (VLDL)-CHOL fractions. In addition, the distributions of the high-density lipoprotein (HDL) CHOL and HDL-TAG fractions were significantly lower (P < 0.01) in dogs with HAC than in healthy dogs. Considering the animals in this study, it was determined that the dogs with HAC differed significantly from the healthy and obese dogs regarding the metabolism of CHOL and TAG, as well as their VLDL and HDL fractions. Similar laboratory findings could allow veterinarians to distinguish obese dogs from those with HAC. In addition, dogs with HAC may be at higher risk for developing metabolic and atherosclerotic complications. PMID- 19286499 TI - Baccharis pteronioides toxicity in livestock and hamsters. AB - Baccharis pteronioides DC has been intermittently associated with livestock poisoning in the southwestern United States. In 2004, nearly 100 cows were reported poisoned by B. pteronioides in southern New Mexico. Initial field studies and postmortem examinations found drought conditions, evidence of B. pteronioides consumption, and a reported mortality of nearly 40%. Because postmortem materials were unsuitable for further examination, plant samples were collected for feeding trials and chemical evaluation. Forty-eight Syrian hamsters (8 weeks old) were randomly divided into 4 groups and dosed with 0, 50, 100, and 200 mg of B. pteronioides for 10 days. After dosing, the hamsters were necropsied; sera were analyzed biochemically; and tissues were collected and evaluated histologically. The hamsters treated with 200 mg and several of the 100 mg animals developed anorexia and diarrhea. These animals developed multiple hemorrhagic infarcts in the liver and kidney, with severe hemorrhagic enteritis. Histologically, the higher-dosed animals had severe necrotizing vasculitis with vascular thrombosis of hepatic and renal vessels. Many glomerular capillaries contained fibrin thrombi. The superficial intestinal and colonic mucosa was necrotic, with extensive hemorrhage and proliferation of luminal bacteria. Lower dosed animals had mild hepatocellular swelling, with proliferation of intestinal and gastric bacteria and yeast. The findings indicate that at high doses, B. pteronioides is toxic to hamsters and produces lesions that are very similar to bacterial endotoxin-produced vasculitis and infarction. Research to purify and identify the toxin, the toxic dose, and mechanism of toxicity are ongoing. PMID- 19286500 TI - Serum acute phase protein concentrations in female dogs with mammary tumors. AB - Acute phase proteins (APPs) are proteins whose concentrations in serum change after any inflammatory stimulus or tissue damage. The aim of the current study was to evaluate 3 positive APPs (C-reactive protein, serum amyloid A, and haptoglobin) and 1 negative APP (albumin) in female dogs with mammary neoplasia. Acute phase proteins were studied in 70 female dogs aged 8-12 years in the following groups: healthy (n = 10); mammary tumors in stages I (n = 19), II (n = 5), III (n = 6), IV (n = 5), and V (n = 7); and with mammary neoplasia plus a concomitant disease (n = 18). In animals with mammary neoplasia, significant increases of positive APPs were only detected in those that had metastasis or a neoplasm with a diameter greater than 5 cm and ulceration. Dogs with mammary neoplasia and a concomitant disease also had high C-reactive protein concentrations. Albumin concentration was decreased in animals with metastasis and with a concomitant disease. The results of the present study indicate that the acute phase response could be stimulated in female dogs with mammary gland tumors because of different factors, such as metastasis, large size of the primary mass, and ulceration or secondary inflammation of the neoplasm. PMID- 19286501 TI - Detection of all eight serotypes of Epizootic hemorrhagic disease virus by real time reverse transcription polymerase chain reaction. AB - Epizootic hemorrhagic disease virus (EHDV) has been associated with bluetongue like disease in cattle. Although U.S. EHDV strains have not been experimentally proven to cause disease in cattle, there is serologic evidence of infection. Differentiation of Bluetongue virus (BTV) and EHDV is necessary because diagnosis of infection caused by these viruses is often confused. The previously developed nested reverse transcription polymerase chain reaction (nRT-PCR) test for indigenous EHDV disease is sensitive and specific, but it is prone to contamination problems. Additionally, the EHDV nRT-PCR only detects 7 of the 8 serotypes. To develop an improved diagnostic test, sequence analysis was performed on 2 conserved target genes; one is highly expressed in infected mammalian cells, whereas the other is highly expressed in infected insect cells. This information was used to develop a rapid EHDV real-time PCR that detects all 8 EHDV serotypes. The EHDV assay did not cross-react with BTV strains and performed similarly to the nRT-PCR tests with archived clinical samples. In addition, it is superior to the nRT-PCR, not only because it is a closed system with fewer cross-contamination problems, but also because it detects all 8 serotypes and is less labor and time intensive. PMID- 19286502 TI - Comparative efficacy of conventional and taqman polymerase chain reaction assays in the detection of capripoxviruses from clinical samples. AB - Sheeppox and goatpox are economically important viral diseases of sheep and goats, respectively. Both diseases are reportable to the World Organization for Animal Health. To implement a control and eradication program for these diseases, a rapid and user-friendly diagnostic tool is imperative for screening. Therefore, in the present study, TaqMan quantitative polymerase chain reaction (qPCR) and conventional PCR assays targeting the DNA polymerase (DNA pol) gene were developed for the detection of Capripoxvirus DNA from clinical specimens of sheep and goats. The 2 assays used different primer sets. Conventional PCR yielded a specific product of 134 bp, whereas qPCR yielded a 180-bp product. The specificity of amplified DNA pol gene products was confirmed by their size and by sequence analysis. The 2 assays were specific for Sheeppox virus and Goatpox virus. However, in comparison to conventional PCR, the qPCR was more rapid, specific, and 100 times more sensitive, with a detection limit as low as 0.042 pg of purified DNA. The qPCR assay was more sensitive (84.05%) than conventional PCR (76.06%) when used on clinical samples (n = 71) from sheep and goats. PMID- 19286503 TI - A polymerase chain reaction strategy for the diagnosis of camelpox. AB - Camelpox is a contagious viral skin disease that is mostly seen in young camels. The disease is caused by the Camelpox virus (CMLV). In the present study, a polymerase chain reaction (PCR) assay based on the C18L gene (encoding ankyrin repeat protein) and a duplex PCR based on the C18L and DNA polymerase (DNA pol) genes were developed. The former assay yields a specific amplicon of 243 bp of the C18L gene, whereas the duplex PCR yields 243- and 96-bp products of the C18L and DNA pol genes, respectively, in CMLV, and only a 96-bp product of the DNA pol gene in other orthopoxviruses. The limit of detection was as low as 0.4 ng of viral DNA. Both PCR assays were employed successfully for the direct detection and differentiation of CMLV from other orthopoxviruses, capripoxviruses, and parapoxviruses in both cell culture samples and clinical material. Furthermore, a highly sensitive SYBR Green dye-based, real-time PCR was optimized for quantitation of CMLV DNA. In the standard curve of the quantitative assay, the melting temperature of the specific amplicon at 77.6 degrees C with peak measured fluorescence in dissociation plot was observed with an efficiency of 102%. To the authors' knowledge, this is the first report to describe a C18L gene-based PCR for specific diagnosis of camelpox infection. PMID- 19286504 TI - Using roquefortine C as a biomarker for penitrem A intoxication. AB - Penitrem A is a well-recognized tremorgenic mycotoxin produced by several Penicillium spp. However, most natural cases of penitrem A intoxication have been associated with Penicillium crustosum. Another Penicillium sp., Penicillium roqueforti, is used for the production of blue cheese and is found in silage and feeds. Penicillium roqueforti produces a mycotoxin, roquefortine C, which is also produced by P. crustosum. In contrast to a tremorgenic syndrome produced by penitrem A, roquefortine C toxicosis is characterized by a paralytic syndrome. Two cases of penitrem A intoxication in dogs are presented to investigate the use of roquefortine C as a biomarker for penitrem A exposure. The vomitus, serum, and urine were analyzed for roquefortine C and penitrem A. Results suggest that roquefortine C can be a sensitive biomarker for penitrem A intoxication. However, the detection of roquefortine C in the absence of penitrem A could merely suggest ingestion of blue cheese or spoilt silage or feed. A review of the literature did not identify any case positive for penitrem A but negative for roquefortine C. In cases in which both mycotoxins were detected, roquefortine C concentration was always higher than penitrem A concentration. In contrast, several cases have been described where the clinical history suggested penitrem A intoxication, but only roquefortine C was detected. In conclusion, roquefortine C can serve as a sensitive biomarker for penitrem A intoxication, but the clinical presentation needs to be considered for proper interpretation of its detection in the absence of penitrem A. PMID- 19286505 TI - Field estimation of the flock-level diagnostic specificity of an enzyme-linked immunosorbent assay for Avian metapneumovirus antibodies in turkeys. AB - Routine serologic testing for Avian metapneumovirus (AMPV) infection of turkey flocks at slaughter is currently being used to monitor changes in the occurrence of AMPV infection in endemic areas and can also be used to detect the emergence of infection in currently unaffected areas. Because of the costs associated with false-positive results, particularly in areas that are free of AMPV infection, there is a need to obtain improved estimates of flock-level specificity (SP). The objective of this study was to estimate flock-level SP of a program to monitor AMPV infection in turkey flocks at processing using a standard enzyme-linked immunosorbent assay (ELISA). A study was carried out in which 37 AMPV-free flocks from 7 Midwest operations were followed serologically. Six percent, 3%, and 0.2% of total samples tested AMPV positive at 8 weeks, 12 weeks, and at processing, respectively. Overall, flock-level SP increased as the cutoff increased and as age increased. Flock-level SP at processing was 97%, if a cutoff of 1 was used (the flock was classified as positive if at least 1 sample tested positive), and 100%, if any other cutoff was used. Administration of antibiotics (P = 0.02) and vaccination for Bordetella avium (P = 0.08) were positively associated with the probability of (false) positive test results. These findings suggest possible cross-reactions with other infections and highlight the need to consider variable diagnostic performance depending on farm conditions. PMID- 19286506 TI - Systemic toxoplasmosis and Gram-negative sepsis in a southern chamois (Rupicapra pyrenaica) from the Pyrenees in northeast Spain. AB - A 6-year-old, male southern chamois (Rupicapra pyrenaica) had an absence of flight response and was captured by hand in the Catalan Pyrenees in northeast Spain. On clinical examination, the animal was in good body condition, and only atrophy of the right eye was observed. Blood samples were collected and hematologic analysis performed, but no alterations were observed. The animal was sent to a Wildlife Rescue Centre, where it developed chronic wasting and died after 32 days in captivity. At necropsy, the animal was cachectic and had edematous, mottled lungs. Histopathologic examination revealed systemic toxoplasmosis and acute Gram-negative septicemia. The protozoan organisms were identified as Toxoplasma gondii based on immunohistochemistry. An indirect fluorescent antibody test was performed, and the animal was positive with an antibody titer of 150. PMID- 19286507 TI - Cotoneaster sp. poisoning in a llama (Lama glama). AB - A case of poisoning with a Cotoneaster sp. is described in a llama (Lama glama). The clinical signs were dyspnea, with congested mucous membranes, chewing, recumbency, and opisthotonos. Several hours after ingestion of the plant, the llama died, despite treatment with sodium thiosulfate and nitrite. Necropsy revealed approximately 676 g of leaves and fruit of Cotoneaster in the first compartment of the stomach. The blood was light red and did not clot. Because Cotoneaster sp. contains low concentrations of cyanogenic glycosides, toxicosis occurs only following massive ingestion of plant material. PMID- 19286508 TI - Naturally occurring sheep-associated malignant catarrhal fever in North American pigs. AB - Two cases of sheep-associated malignant catarrhal fever (MCF) in pigs were diagnosed on a small farm in New York, and in Kentucky. In both cases, the initial diagnosis was based on histopathological changes representing typical lymphoproliferative vasculitis in multiple tissues of the affected pigs. Ovine herpesvirus 2 (OvHV-2) DNA was detected by polymerase chain reaction in the tissues of affected pigs in both cases. The amplified sequences were identical between the clinically affected pigs and the OvHV-2-infected sheep. Additional virological and bacteriological examination showed that the affected pigs were negative for agents that cause Aujeszky's disease, classical swine fever, porcine enterovirus, and rabies. An antibody against a conserved epitope among MCF viruses was detected in 1 clinically affected pig and 2 unaffected cohort pigs, as well as in all the associated sheep. To the authors' knowledge, this is the first report of porcine MCF in North America, and suggests that OvHV-2 is associated with clinical MCF in pigs. PMID- 19286509 TI - Spinal lymphoma and pulmonary filariasis in a pet domestic rabbit (Oryctolagus cuniculus domesticus). AB - Spinal lymphoma and concurrent pulmonary filariasis are reported in a pet rabbit. The rabbit presented for pelvic limb paralysis resulting from extradural spinal lymphoma, presumably rising from the body of the sixth lumbar vertebra. The neoplasm was subsequently immunophenotyped as a B-cell lymphoma. Pulmonary filariasis was an incidental finding at necropsy. PMID- 19286510 TI - Infection with Ovine herpesvirus 2 in Norwegian herds with a history of previous outbreaks of malignant catarrhal fever. AB - Infection with Ovine herpesvirus 2 (OvHV-2) in healthy cattle, swine, sheep, and goats was investigated on 43 selected Norwegian farms; of which, 41 (95%) had experienced outbreaks of malignant catarrhal fever (MCF) in cattle and/or swine during the preceding 5 years. Two of the farms had no history of MCF and were included for control purposes. Blood samples from 384 cattle, 40 sows, 75 sheep, and 4 goats were examined for OvHV-2 by polymerase chain reaction assay (PCR) and for antibodies using a competitive inhibition enzyme-linked immunosorbent assay (ciELISA). All samples were also tested for antibodies reactive to Alcelaphine herpesvirus 1 with an indirect fluorescent antibody test (IFAT). All but 4 of the sheep and all 4 goats tested positive with 1 or more of the tests. Eighty-nine (25%) of the cattle and 17 (43%) of the swine on the farms with previous MCF outbreaks tested positive with 1 or more of the tests. On 22 of the farms, at least 1 bovine tested positive with ciELISA and/or PCR, whereas 8 other farms had test-positive cattle with IFAT only. The 2 control farms yielded no positive results with any of the tests. Four of the farms had swine that tested positive with PCR, but none with ciELISA, whereas 4 other farms had test-positive swine with IFAT only. The prevalence of infection in cattle and swine seemed not to be influenced either by their age or the degree of contact with the sheep and goats. PMID- 19286511 TI - Pituitary acidophil macroadenoma in a pygmy goat (Capra hircus hircus). AB - This report describes a pituitary acidophil macroadenoma in a goat. Antemortem clinical findings included hypothermia and rumen stasis. Clinicopathologic findings included refractory hypoglycemia, low total thyroxin and insulin concentrations, elevated bile acid concentration, and hyposthenuria. In addition to the pituitary macroadenoma, bilateral atrophy of the zona reticularis of the adrenal glands was observed histologically. PMID- 19286512 TI - Isolation of Campylobacter fetus subspecies fetus in a two-year-old quarterhorse with chronic diarrhea of an undetermined etiology. AB - A 2-year-old quarterhorse was evaluated for chronic diarrhea and weight loss of 5 weeks duration after numerous diagnostic tests failed to identify an underlying cause. Historically, the horse was housed at pasture where human household waste vehicles were routinely cleaned and the effluent could run onto the field. Physical examination revealed poor body condition and frequent high-volume diarrhea. Diagnostic testing for Salmonella spp., endoparasites, Cryptosporidium spp., Clostridium spp., and diffuse infiltrative bowel disease were negative. Rectal tissue histopathology failed to identify Mycobacterium spp., spirochetal organisms, or submucosal infiltration with cells. Rectal tissue biopsy and a fresh fecal sample identified numerous Campylobacter organisms with microaerobic culture. Molecular testing revealed the species as Campylobacter fetus subsp. fetus as a possible organism associated with large bowel disease in this filly. The organism was sensitive to fluoroquinolone antimicrobials in vitro. The filly responded transiently to therapy, forming discrete fecal balls after 72 hr of treatment. At 5 months follow-up, the horse had gained weight, was alert and responsive, but reverted back to having soft "cow-pie" feces. Reculture of the feces at 9 months failed to identify any Campylobacter organisms. To the authors' knowledge, this is the first report to identify C. fetus subsp. fetus from feces and tissue of a horse with the use of molecular methods. This organism could play a role in the etiology of chronic diarrhea in horses. PMID- 19286513 TI - Pneumonia from Angiostrongylus vasorum infection in a red panda (Ailurus fulgens fulgens). AB - A 9-year-old, male, captive red panda (Ailurus fulgens fulgens) in an urban zoo in the United Kingdom presented with respiratory distress and weight loss. The animal was euthanatized, and a postmortem examination was performed. The lungs were diffusely consolidated with extensive mineralization. Microscopically, there was extensive obliteration of normal pulmonary architecture by sheets and coalescing nodules of partially mineralized fibrous tissue and granulomatous inflammation centered on large numbers of nematode larvae and eggs. First stage nematode larvae were isolated from lung tissue and were characterized as Angiostrongylus vasorum on the basis of their morphology and sequencing of the 18S ribosomal RNA gene and the entire second internal transcribed spacer. Although A. vasorum has previously been reported in red pandas in a zoological collection in Denmark, this study is the first reported case in the United Kingdom and occurs against a background of geographical spread and increased incidence of disease in domestic and wild canids. Angiostrongylus vasorum should be considered a differential diagnosis for respiratory disease in the red panda and taken into account when planning parasite and pest control programs for zoological collections. PMID- 19286514 TI - Isolation of Propionibacterium acnes from a case of placentitis and abortion in a cow. AB - On the basis of the scarcity of reports in the veterinary literature, it appears that Propionibacterium spp. are rarely associated with disease or isolated from cattle tissues. Recently, Propionibacterium spp. has been associated with multifocal abscessation in cattle. This report describes a case of necrosuppurative placentitis and abortion in an adult Holstein cow. Numerous colonies of small, pleomorphic, Gram-positive, rod-shaped bacteria were observed within the fibrin lattice associated with placental lesions and within the fetal atelectatic lung. Propionibacterium acnes was isolated in high numbers from the placenta, fetal lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with propionibacteria in a cow. PMID- 19286515 TI - Feline intestinal T-cell lymphoma: assessment of morphologic and kinetic features in 30 cases. AB - In this study, 30 feline intestinal T-cell lymphomas (ITCLs) from 77 cats with gastrointestinal lymphoma were evaluated. Neoplastic lesions were composed predominantly of small (n = 21) or medium to large (n = 9) anaplastic cells. Different patterns of tumor growth were observed. A starry-sky pattern was evident in 7 cases (23.3%), fibrosis in 18 cases (60%), and neovascularization in 19 cases (63.3%). The cell proliferation index (assessed by MIB1 [mindbomb homolog 1] immunohistochemistry) ranged from 0.21% to 66.91%. Mean MIB1 index was 3.49% within the first 33rd percentile, 18.31% within the second 33rd percentile, and 40.16% within the third 33rd percentile (P < 0.000001). Microscopic and kinetic features provided evidence that ITCL in cats exhibits a spectrum in cytological composition and growth patterns that could putatively reflect differences in biologic behavior. PMID- 19286516 TI - Metastatic uterine adenocarcinoma and hepatic lipomatosis in a llama (Lama glama). AB - A 6-year-old, female llama (Lama glama) presented with progressive anorexia and ascites. Postmortem examination revealed an infiltrative uterine adenocarcinoma with widespread metastases. The neoplasm completely replaced and infiltrated the myometrium of the uterine body and cervix and metastasized largely to the serosal surfaces of the peritoneal cavity. Histopathology identified a highly invasive growth of neoplastic cells in solid packets or tubular arrangements and marked fibroplasia. No bovine or llama papillomavirus DNA was detected intralesionally by polymerase chain reaction. PMID- 19286517 TI - Sarcocystis falcatula-associated encephalitis in a free-ranging great horned owl (Bubo virginianus). AB - A great horned owl (Bubo virginianus) was admitted to a rehabilitation clinic with severe neurologic signs that were unresponsive to supportive care. The animal was euthanatized because of a poor prognosis. Marked granulomatous encephalitis with focal brainstem malacia was detected microscopically. The brainstem was the most severely affected brain location and the only place in which schizonts and merozoites, morphologically compatible with Sarcocystis spp., were detected. Immunohistochemistry with the use of polyclonal antisera indicated the presence of Sarcocystis falcatula. The species identification of the protozoa as S. falcatula was confirmed by polymerase chain reaction. To the author's knowledge, this is the first report of spontaneous S. falcatula-associated encephalitis in a great horned owl. PMID- 19286518 TI - Circadian control of the NAD+ salvage pathway by CLOCK-SIRT1. AB - Many metabolic and physiological processes display circadian oscillations. We have shown that the core circadian regulator, CLOCK, is a histone acetyltransferase whose activity is counterbalanced by the nicotinamide adenine dinucleotide (NAD+)-dependent histone deacetylase SIRT1. Here we show that intracellular NAD+ levels cycle with a 24-hour rhythm, an oscillation driven by the circadian clock. CLOCK:BMAL1 regulates the circadian expression of NAMPT (nicotinamide phosphoribosyltransferase), an enzyme that provides a rate-limiting step in the NAD+ salvage pathway. SIRT1 is recruited to the Nampt promoter and contributes to the circadian synthesis of its own coenzyme. Using the specific inhibitor FK866, we demonstrated that NAMPT is required to modulate circadian gene expression. Our findings in mouse embryo fibroblasts reveal an interlocked transcriptional-enzymatic feedback loop that governs the molecular interplay between cellular metabolism and circadian rhythms. PMID- 19286519 TI - In vivo analysis of dendritic cell development and homeostasis. AB - Dendritic cells (DCs) in lymphoid tissue arise from precursors that also produce monocytes and plasmacytoid DCs (pDCs). Where DC and monocyte lineage commitment occurs and the nature of the DC precursor that migrates from the bone marrow to peripheral lymphoid organs are unknown. We show that DC development progresses from the macrophage and DC precursor to common DC precursors that give rise to pDCs and classical spleen DCs (cDCs), but not monocytes, and finally to committed precursors of cDCs (pre-cDCs). Pre-cDCs enter lymph nodes through and migrate along high endothelial venules and later disperse and integrate into the DC network. Further cDC development involves cell division, which is controlled in part by regulatory T cells and fms-like tyrosine kinase receptor-3. PMID- 19286520 TI - Local DNA topography correlates with functional noncoding regions of the human genome. AB - The three-dimensional molecular structure of DNA, specifically the shape of the backbone and grooves of genomic DNA, can be dramatically affected by nucleotide changes, which can cause differences in protein-binding affinity and phenotype. We developed an algorithm to measure constraint on the basis of similarity of DNA topography among multiple species, using hydroxyl radical cleavage patterns to interrogate the solvent-accessible surface area of DNA. This algorithm found that 12% of bases in the human genome are evolutionarily constrained-double the number detected by nucleotide sequence-based algorithms. Topography-informed constrained regions correlated with functional noncoding elements, including enhancers, better than did regions identified solely on the basis of nucleotide sequence. These results support the idea that the molecular shape of DNA is under selection and can identify evolutionary history. PMID- 19286521 TI - Solar power wires based on organic photovoltaic materials. AB - Organic photovoltaics in a flexible wire format has potential advantages that are described in this paper. A wire format requires long-distance transport of current that can be achieved only with conventional metals, thus eliminating the use of transparent oxide semiconductors. A phase-separated, photovoltaic layer, comprising a conducting polymer and a fullerene derivative, is coated onto a thin metal wire. A second wire, coated with a silver film, serving as the counter electrode, is wrapped around the first wire. Both wires are encased in a transparent polymer cladding. Incident light is focused by the cladding onto to the photovoltaic layer even when it is completely shadowed by the counter electrode. Efficiency values of the wires range from 2.79% to 3.27%. PMID- 19286522 TI - Scientist citizens. PMID- 19286523 TI - Biomedical research. A first step in relaxing restrictions on stem cell research. PMID- 19286524 TI - Research funding. England spreads its funds widely, sparking debate. PMID- 19286525 TI - Biomedical research. Humane Society launches offensive to ban invasive chimp research. PMID- 19286526 TI - Solar physics. Report puts NASA's solar program under a cloud. PMID- 19286527 TI - Biosecurity. Paul Keim on his life with the FBI during the anthrax investigation. Interview by Yudhijit Bhattacharjee. PMID- 19286528 TI - China. Biologists muscle up with major new protein facilities. PMID- 19286529 TI - Planetary science. In dune map, Titan's winds seem to blow backward. PMID- 19286530 TI - Paleoanthropology. Ice Age no barrier to 'Peking Man'. PMID- 19286532 TI - Geology. How much coal remains? PMID- 19286533 TI - Neuroscience. A memorable device. PMID- 19286534 TI - Nuclear physics. Fathoming matter's heart unbound. PMID- 19286535 TI - A national initiative for social participation. PMID- 19286536 TI - Indian neutrino detector: the elephant in the room. PMID- 19286537 TI - Indian neutrino detector: environmental costs. PMID- 19286538 TI - Comment on "Detection, stimulation, and inhibition of neuronal signals with high density nanowire transistor arrays". AB - Patolsky et al. (Reports, 25 August 2006, p. 1100) used silicon nanowires to record action potentials in rat neuronal axons and found increases in conductance of about 85 nanosiemens. We point out that the data correspond to voltage changes of about -85 millivolts on the nanowire and that conceivable mechanisms of axon nanowire interaction lead to signals that are opposite in sign or smaller by orders of magnitude. PMID- 19286539 TI - Resource policy. Wood energy in America. PMID- 19286540 TI - Climate change. Shifting westerlies. PMID- 19286541 TI - Chemistry. Inducing chirality with circularly polarized light. PMID- 19286542 TI - Biochemistry. The evolution of ribozyme chemistry. PMID- 19286543 TI - Mathematics. Emergence of connectivity in networks. PMID- 19286544 TI - Plant science. Paternal patterning cue. PMID- 19286545 TI - Circadian rhythms. Linking the loops. PMID- 19286546 TI - Time-variable deformation in the New Madrid seismic zone. AB - New geodetic measurements show that the New Madrid is currently deforming too slowly, if at all, to account for large earthquakes in the region over the past 5000 years. This result, together with increasing evidence for temporal clustering and spatial migration of earthquake sequences in continental interiors, indicates that either tectonic loading rates or fault properties vary over a few thousand years. PMID- 19286547 TI - Wind-driven upwelling in the Southern Ocean and the deglacial rise in atmospheric CO2. AB - Wind-driven upwelling in the ocean around Antarctica helps regulate the exchange of carbon dioxide (CO2) between the deep sea and the atmosphere, as well as the supply of dissolved silicon to the euphotic zone of the Southern Ocean. Diatom productivity south of the Antarctic Polar Front and the subsequent burial of biogenic opal in underlying sediments are limited by this silicon supply. We show that opal burial rates, and thus upwelling, were enhanced during the termination of the last ice age in each sector of the Southern Ocean. In the record with the greatest temporal resolution, we find evidence for two intervals of enhanced upwelling concurrent with the two intervals of rising atmospheric CO2 during deglaciation. These results directly link increased ventilation of deep water to the deglacial rise in atmospheric CO2. PMID- 19286548 TI - Explosive percolation in random networks. AB - Networks in which the formation of connections is governed by a random process often undergo a percolation transition, wherein around a critical point, the addition of a small number of connections causes a sizable fraction of the network to suddenly become linked together. Typically such transitions are continuous, so that the percentage of the network linked together tends to zero right above the transition point. Whether percolation transitions could be discontinuous has been an open question. Here, we show that incorporating a limited amount of choice in the classic Erdos-Renyi network formation model causes its percolation transition to become discontinuous. PMID- 19286549 TI - The initial stages of template-controlled CaCO3 formation revealed by cryo-TEM. AB - Biogenic calcium carbonate forms the inorganic component of seashells, otoliths, and many marine skeletons, and its formation is directed by an ordered template of macromolecules. Classical nucleation theory considers crystal formation to occur from a critical nucleus formed by the assembly of ions from solution. Using cryotransmission electron microscopy, we found that template-directed calcium carbonate formation starts with the formation of prenucleation clusters. Their aggregation leads to the nucleation of amorphous nanoparticles in solution. These nanoparticles assemble at the template and, after reaching a critical size, develop dynamic crystalline domains, one of which is selectively stabilized by the template. Our findings have implications for template-directed mineral formation in biological as well as in synthetic systems. PMID- 19286550 TI - Self-repairing oxetane-substituted chitosan polyurethane networks. AB - Polyurethanes have many properties that qualify them as high-performance polymeric materials, but they still suffer from mechanical damage. We report the development of polyurethane networks that exhibit self-repairing characteristics upon exposure to ultraviolet light. The network consists of an oxetane substituted chitosan precursor incorporated into a two-component polyurethane. Upon mechanical damage of the network, four-member oxetane rings open to create two reactive ends. When exposed to ultraviolet light, chitosan chain scission occurs, which forms crosslinks with the reactive oxetane ends, thus repairing the network. These materials are capable of repairing themselves in less than an hour and can be used in many coatings applications, ranging from transportation to packaging or fashion and biomedical industries. PMID- 19286551 TI - Nonporous organic solids capable of dynamically resolving mixtures of diiodoperfluoroalkanes. AB - Halogen bonding has increasingly facilitated the assembly of diverse host-guest solids. Here, we show that a well-known class of organic salts, bis(trimethylammonium) alkane diiodides, can reversibly encapsulate alpha,omega diiodoperfluoroalkanes (DIPFAs) through intermolecular interactions between the host's I- anions and the guest's terminal iodine substituents. The process is highly selective for the fluorocarbon that forms an I-...I(CF2)mI...I- superanion that is matched in length to the chosen dication. DIPFAs that are 2 to 12 carbons in length (common industrial intermediates) can thereby be isolated from mixtures by means of crystallization from solution upon addition of the dissolved size matched ionic salt. The solid-state salts can also selectively capture the DIPFAs from the vapor phase, yielding the same product formed from solution despite a lack of porosity of the starting lattice structure. Heating liberates the DIPFAs and regenerates the original salt lattice, highlighting the practical potential for the system in separation applications. PMID- 19286552 TI - Time-resolved molecular frame dynamics of fixed-in-space CS2 molecules. AB - Random orientation of molecules within a sample leads to blurred observations of chemical reactions studied from the laboratory perspective. Methods developed for the dynamic imaging of molecular structures and processes struggle with this, as measurements are optimally made in the molecular frame. We used laser alignment to transiently fix carbon disulfide molecules in space long enough to elucidate, in the molecular reference frame, details of ultrafast electronic-vibrational dynamics during a photochemical reaction. These three-dimensional photoelectron imaging results, combined with ongoing efforts in molecular alignment and orientation, presage a wide range of insights obtainable from time-resolved studies in the molecular frame. PMID- 19286553 TI - Clear sky visibility has decreased over land globally from 1973 to 2007. AB - Visibility in the clear sky is reduced by the presence of aerosols, whose types and concentrations have a large impact on the amount of solar radiation that reaches Earth's surface. Here we establish a global climatology of inverse visibilities over land from 1973 to 2007 and interpret it in terms of changes in aerosol optical depth and the consequent impacts on incident solar radiation. The aerosol contribution to "global dimming," first reported in terms of strong decreases in measured incident solar radiation up to the mid-1980s, has monotonically increased over the period analyzed. Since that time, visibility has increased over Europe, consistent with reported European "brightening," but has decreased substantially over south and east Asia, South America, Australia, and Africa, resulting in net global dimming over land. PMID- 19286554 TI - Recent changes in phytoplankton communities associated with rapid regional climate change along the western Antarctic Peninsula. AB - The climate of the western shelf of the Antarctic Peninsula (WAP) is undergoing a transition from a cold-dry polar-type climate to a warm-humid sub-Antarctic-type climate. Using three decades of satellite and field data, we document that ocean biological productivity, inferred from chlorophyll a concentration (Chl a), has significantly changed along the WAP shelf. Summertime surface Chl a (summer integrated Chl a approximately 63% of annually integrated Chl a) declined by 12% along the WAP over the past 30 years, with the largest decreases equatorward of 63 degrees S and with substantial increases in Chl a occurring farther south. The latitudinal variation in Chl a trends reflects shifting patterns of ice cover, cloud formation, and windiness affecting water-column mixing. Regional changes in phytoplankton coincide with observed changes in krill (Euphausia superba) and penguin populations. PMID- 19286555 TI - A recessive mutation in the APP gene with dominant-negative effect on amyloidogenesis. AB - beta-Amyloid precursor protein (APP) mutations cause familial Alzheimer's disease with nearly complete penetrance. We found an APP mutation [alanine-673-->valine 673 (A673V)] that causes disease only in the homozygous state, whereas heterozygous carriers were unaffected, consistent with a recessive Mendelian trait of inheritance. The A673V mutation affected APP processing, resulting in enhanced beta-amyloid (Abeta) production and formation of amyloid fibrils in vitro. Co-incubation of mutated and wild-type peptides conferred instability on Abeta aggregates and inhibited amyloidogenesis and neurotoxicity. The highly amyloidogenic effect of the A673V mutation in the homozygous state and its anti amyloidogenic effect in the heterozygous state account for the autosomal recessive pattern of inheritance and have implications for genetic screening and the potential treatment of Alzheimer's disease. PMID- 19286556 TI - Structure of the anaphase-promoting complex/cyclosome interacting with a mitotic checkpoint complex. AB - Once all chromosomes are connected to the mitotic spindle (bioriented), anaphase is initiated by the protein ubiquitylation activity of the anaphase-promoting complex/cyclosome (APC/C) and its coactivator Cdc20 (APC/C(Cdc20)). Before chromosome biorientation, anaphase is delayed by a mitotic checkpoint complex (MCC) that inhibits APC/C(Cdc20). We used single-particle electron microscopy to obtain three-dimensional models of human APC/C in various functional states: bound to MCC, to Cdc20, or to neither (apo-APC/C). These experiments revealed that MCC associates with the Cdc20 binding site on APC/C, locks the otherwise flexible APC/C in a "closed" state, and prevents binding and ubiquitylation of a wide range of different APC/C substrates. These observations clarify the structural basis for the inhibition of APC/C by spindle checkpoint proteins. PMID- 19286557 TI - A functional genomics approach reveals CHE as a component of the Arabidopsis circadian clock. AB - Transcriptional feedback loops constitute the molecular circuitry of the plant circadian clock. In Arabidopsis, a core loop is established between CCA1 and TOC1. Although CCA1 directly represses TOC1, the TOC1 protein has no DNA binding domains, which suggests that it cannot directly regulate CCA1. We established a functional genomic strategy that led to the identification of CHE, a TCP transcription factor that binds specifically to the CCA1 promoter. CHE is a clock component partially redundant with LHY in the repression of CCA1. The expression of CHE is regulated by CCA1, thus adding a CCA1/CHE feedback loop to the Arabidopsis circadian network. Because CHE and TOC1 interact, and CHE binds to the CCA1 promoter, a molecular linkage between TOC1 and CCA1 gene regulation is established. PMID- 19286558 TI - Paternal control of embryonic patterning in Arabidopsis thaliana. AB - The YODA (YDA) mitogen-activated protein kinase pathway promotes elongation of the Arabidopsis zygote and development of its basal daughter cell into the extra embryonic suspensor. Here, we show that the interleukin-1 receptor-associated kinase (IRAK)/Pelle-like kinase gene SHORT SUSPENSOR (SSP) regulates this pathway through a previously unknown parent-of-origin effect. SSP transcripts are produced in mature pollen but do not appear to be translated. Instead, they are delivered via the sperm cells to the zygote and the endosperm, where SSP protein transiently accumulates. Ectopic expression of SSP protein in the leaf epidermis is sufficient to activate YDA-dependent signaling. We propose that SSP protein produced from paternal transcripts upon fertilization triggers zygotic YDA activity, providing an essential temporal cue for the regulation of the asymmetric first division. PMID- 19286559 TI - Preferential generation of follicular B helper T cells from Foxp3+ T cells in gut Peyer's patches. AB - Most of the immunoglobulin A (IgA) in the gut is generated by B cells in the germinal centers of Peyer's patches through a process that requires the presence of CD4+ follicular B helper T(TFH) cells. The nature of these T(FH) cells in Peyer's patches has been elusive. Here, we demonstrate that suppressive Foxp3+CD4+ T cells can differentiate into TFH cells in mouse Peyer's patches. The conversion of Foxp3+ T cells into TFH cells requires the loss of Foxp3 expression and subsequent interaction with B cells. Thus, environmental cues present in gut Peyer's patches promote the selective differentiation of distinct helper T cell subsets, such as TFH cells. PMID- 19286560 TI - Selective erasure of a fear memory. AB - Memories are thought to be encoded by sparsely distributed groups of neurons. However, identifying the precise neurons supporting a given memory (the memory trace) has been a long-standing challenge. We have shown previously that lateral amygdala (LA) neurons with increased cyclic adenosine monophosphate response element-binding protein (CREB) are preferentially activated by fear memory expression, which suggests that they are selectively recruited into the memory trace. We used an inducible diphtheria-toxin strategy to specifically ablate these neurons. Selectively deleting neurons overexpressing CREB (but not a similar portion of random LA neurons) after learning blocked expression of that fear memory. The resulting memory loss was robust and persistent, which suggests that the memory was permanently erased. These results establish a causal link between a specific neuronal subpopulation and memory expression, thereby identifying critical neurons within the memory trace. PMID- 19286561 TI - Human substantia nigra neurons encode unexpected financial rewards. AB - The brain's sensitivity to unexpected outcomes plays a fundamental role in an organism's ability to adapt and learn new behaviors. Emerging research suggests that midbrain dopaminergic neurons encode these unexpected outcomes. We used microelectrode recordings during deep brain stimulation surgery to study neuronal activity in the human substantia nigra (SN) while patients with Parkinson's disease engaged in a probabilistic learning task motivated by virtual financial rewards. Based on a model of the participants' expected reward, we divided trial outcomes into expected and unexpected gains and losses. SN neurons exhibited significantly higher firing rates after unexpected gains than unexpected losses. No such differences were observed after expected gains and losses. This result provides critical support for the hypothesized role of the SN in human reinforcement learning. PMID- 19286562 TI - RSY-1 is a local inhibitor of presynaptic assembly in C. elegans. AB - As fundamental units of neuronal communication, chemical synapses are composed of presynaptic and postsynaptic specializations that form at specific locations with defined shape and size. Synaptic assembly must be tightly regulated to prevent overgrowth of the synapse size and number, but the molecular mechanisms that inhibit synapse assembly are poorly understood. We identified regulator of synaptogenesis-1 (RSY-1) as an evolutionarily conserved molecule that locally antagonized presynaptic assembly. The loss of RSY-1 in Caenorhabditis elegans led to formation of extra synapses and recruitment of excessive synaptic material to presynaptic sites. RSY-1 directly interacted with and negatively regulated SYD 2/liprin-alpha, a master assembly molecule that recruits numerous synaptic components to presynaptic sites. RSY-1 also bound and regulated SYD-1, a synaptic protein required for proper functioning of SYD-2. Thus, local inhibitory mechanisms govern synapse formation. PMID- 19286564 TI - Tracking early autoimmune disease by bioluminescent imaging of NF-kappaB activation reveals pathology in multiple organ systems. AB - It is desirable to have an early and sensitive detection marker of autoimmune disease in intact animals. Nuclear factor (NF)-kappaB is a transcription factor that is associated with inflammatory responses and immune disorders. Previously, we demonstrated that so-called idiotypic-driven T-B cell collaboration in mice doubly transgenic for paired immunoglobulin and T cell receptor transgenes resulted in a systemic autoimmune disease with systemic lupus erythematosus-like features. Here, we investigated NF-kappaB activation by including an NF-kappaB responsive luciferase reporter transgene in this animal model. Triply transgenic mice developed bioluminescence signals from diseased organs before onset of clinical symptoms and autoantibody production, and light emissions correlated with disease progression. Signals were obtained from secondary lymphoid organs, inflamed intestines, skin lesions, and arthritic joints. Moreover, bioluminescence imaging and immunohistochemistry demonstrated that a minority of mice suffered from an autoimmune disease of the small intestine, in which light emissions correlated with antibodies against tissue transglutaminase and gliadin. Detection of luciferase by immunohistochemistry revealed NF-kappaB activation in collaborating B and T cells, as well as in macrophages. These results demonstrate that bioluminescent in vivo imaging of NF-kappaB activation can be used for early and sensitive detection of autoimmune disease in an experimental mouse model, offering new possibilities for the evaluation of anti-inflammatory drugs. PMID- 19286565 TI - The bacterial virulence factor lymphostatin compromises intestinal epithelial barrier function by modulating rho GTPases. AB - Lymphocyte inhibitory factor A (lifA) in Citrobacter rodentium encodes the large toxin lymphostatin, which contains two enzymatic motifs associated with bacterial pathogenesis, a glucosyltransferase and a protease. Our aim was to determine the effects of each lymphostatin motif on intestinal epithelial-barrier function. In frame mutations of C. rodentium lifA glucosyltransferase (CrGlM21) and protease (CrPrM5) were generated by homologous recombination. Infection of both model intestinal epithelial monolayers and mice with C. rodentium wild type resulted in compromised epithelial barrier function and mislocalization of key intercellular junction proteins in the tight junction and adherens junction. In contrast, CrGlM21 was impaired in its ability to reduce barrier function and influenced the tight junction proteins ZO-1 and occludin. CrPrM5 demonstrated decreased effects on the adherens junction proteins beta-catenin and E-cadherin. Analysis of the mechanisms revealed that C. rodentium wild type differentially influenced Rho GTPase activation, suppressed Cdc42 activation, and induced Rho GTPase activation. CrGlM21 lost its suppressive effects on Cdc42 activation, whereas CrPrM5 was unable to activate Rho signaling. Rescue experiments using constitutively active Cdc42 or C3 exotoxin to inhibit Rho GTPase supported a role of Rho GTPases in the epithelial barrier compromise induced by C. rodentium. Taken together, our results suggest that lymphostatin is a bacterial virulence factor that contributes to the disruption of intestinal epithelial-barrier function via the modulation of Rho GTPase activities. PMID- 19286566 TI - The long pentraxin PTX3 is crucial for tissue inflammation after intestinal ischemia and reperfusion in mice. AB - The pentraxin superfamily is a group of evolutionarily conserved proteins that play important roles in the immune system. The long pentraxin PTX3 protein was originally described as able to be induced by pro-inflammatory stimuli in a variety of cell types. In this study, we evaluated the phenotype of Ptx3(-/-) mice subjected to ischemia followed by reperfusion of the superior mesenteric artery. In reperfused wild-type mice, there was significant local and remote injury as demonstrated by increases in vascular permeability, neutrophil influx, nuclear factor-kappaB activation, and production of CXCL1 and tumor necrosis factor-alpha. PTX3 levels were elevated in both serum and intestine after reperfusion. In Ptx3(-/-) mice, local and remote tissue injury was inhibited, and there were decreased nuclear factor-kappaB translocation and cytokine production. Intestinal architecture was preserved, and there were decreased neutrophil influx and significant prevention of lethality in Ptx3(-/-) mice as well. PTX3 given intravenously before reperfusion reversed the protection observed in Ptx3(-/-) mice in a dose-dependent manner, and PTX3 administration significantly worsened tissue injury and lethality in wild-type mice. In conclusion, our studies demonstrate a major role for PTX3 in determining acute reperfusion-associated inflammation, tissue injury, and lethality and suggest the soluble form of this molecule is active in this system. Therapeutic blockade of PTX3 action may be useful in the control of the injuries associated with severe ischemia and reperfusion syndromes. PMID- 19286567 TI - GLUT1 expression is increased in hepatocellular carcinoma and promotes tumorigenesis. AB - Accelerated glycolysis is one of the biochemical characteristics of cancer cells. The glucose transporter isoform 1 (GLUT1) gene encodes a key rate-limiting factor in glucose transport into cancer cells. However, its expression level and functional significance in hepatocellular cancer (HCC) are still disputed. Therefore, we aimed to analyze the expression and function of the GLUT1 gene in cases of HCC. We found significantly higher GLUT1 mRNA expression levels in HCC tissues and cell lines compared with primary human hepatocytes and matched nontumor tissue. Immunohistochemical analysis of a tissue microarray of 152 HCC cases revealed a significant correlation between Glut1 protein expression levels and a higher Ki-67 labeling index, advanced tumor stages, and poor differentiation. Accordingly, suppression of GLUT1 expression by siRNA significantly impaired both the growth and migratory potential of HCC cells. Furthermore, inhibition of GLUT1 expression reduced both glucose uptake and lactate secretion. Hypoxic conditions further increased GLUT1 expression levels in HCC cells, and this induction was dependent on the activation of the transcription factor hypoxia-inducible factor-1alpha. In summary, our findings suggest that increased GLUT1 expression levels in HCC cells functionally affect tumorigenicity, and thus, we propose GLUT1 as an innovative therapeutic target for this highly aggressive tumor. PMID- 19286568 TI - Microtubule depolymerization suppresses alpha-synuclein accumulation in a mouse model of multiple system atrophy. AB - Multiple system atrophy (MSA) is a neurodegenerative disease caused by an accumulation of alpha-synuclein (alpha-syn) in oligodendrocytes. Little is known about the cellular mechanisms by which alpha-syn accumulation causes neuronal degeneration in MSA. Our previous research, however, revealed that in a mouse model of MSA, oligodendrocytic inclusions of alpha-syn induced neuronal accumulation of alpha-syn, as well as progressive neuronal degeneration. Here we identify the mechanisms that underlie neuronal accumulation of alpha-syn in a mouse MSA model. We found that the alpha-syn protein binds to beta-III tubulin in microtubules to form an insoluble complex. The insoluble alpha-syn complex progressively accumulates in neurons and leads to neuronal dysfunction. Furthermore, we demonstrated that the neuronal accumulation of insoluble alpha syn is suppressed by treatment with a microtubule depolymerizing agent. The underlying pathological process appeared to also be inhibited by this treatment, providing promise for future therapeutic approaches. PMID- 19286569 TI - Essential role for macrophage migration inhibitory factor in gastritis induced by Helicobacter pylori. AB - Macrophage migration inhibitory factor (MIF) is an upstream regulator of immune and inflammatory responses; however, its role in Helicobacter pylori (HP) associated gastritis remains unknown. We infected MIF knockout (KO) and wild-type mice with SS1 HP and found that 2 weeks after infection, MIF and its receptor CD74 were markedly up-regulated in wild-type mice. This up-regulation preceded the up-regulation of both tumor necrosis factor-alpha and intercellular adhesion molecule-1, as well as the development of moderate gastritis at 8 weeks, as determined by a significant infiltration of neutrophils, T cells, and macrophages. In contrast, KO mice were protected against HP-induced gastritis by preventing the up-regulation of CD74 and Th1-mediated immune injury, including a reduction in the Th1 transcriptional factor T-bet and the expression of interferon-gamma. Additionally, inhibition of skin delayed type hypersensitivity reactions to HP antigens in KO mice also suggested a critical role for MIF in cell-mediated injury. A regulatory role for MIF in Th1-immune responses was further demonstrated by the finding that antigen-primed CD4(+) T cells lacking MIF failed to differentiate into the Th1 phenotype; these cells were instead promoted to Th2 differentiation after challenge with HP antigen in vitro. Results from this study indicated that inhibition of HP-induced innate immune responses and Th1-mediated immune injury may be the key mechanisms by which KO mice failed to develop gastritis after HP infection. PMID- 19286570 TI - Regulation of prostate cancer progression by galectin-3. AB - Galectin-3, a beta-galactoside-binding protein, has been implicated in a variety of biological functions including cell proliferation, apoptosis, angiogenesis, tumor progression, and metastasis. The present study was undertaken to understand the role of galectin-3 in the progression of prostate cancer. Immunohistochemical analysis of galectin-3 expression revealed that galectin-3 was cleaved during the progression of prostate cancer. Galectin-3 knockdown by small interfering RNA (siRNA) was associated with reduced cell migration, invasion, cell proliferation, anchorage-independent colony formation, and tumor growth in the prostates of nude mice. Galectin-3 knockdown in human prostate cancer PC3 cells led to cell-cycle arrest at G(1) phase, up-regulation of nuclear p21, and hypophosphorylation of the retinoblastoma tumor suppressor protein (pRb), with no effect on cyclin D1, cyclin E, cyclin-dependent kinases (CDK2 and CDK4), and p27 protein expression levels. The data obtained here implicate galectin-3 in prostate cancer progression and suggest that galectin-3 may serve as both a diagnostic marker and therapeutic target for future disease treatments. PMID- 19286571 TI - Endurance exercise is protective for mice with mitochondrial myopathy. AB - Defects in the mitochondrial ATP-generating system are one of the most commonly inherited neurological disorders, but they remain without treatment. We have recently shown that modulation of the peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) level in skeletal muscle of a mitochondrial myopathy mouse model offers a therapeutic approach. Here we analyzed if endurance exercise, which is known to be associated with an increased PGC-1alpha level in muscle, offers the same beneficial effect. We subjected male and female mice that develop a severe mitochondrial myopathy due to a cytochrome-c oxidase deficiency at 3 mo of age to endurance exercise training and monitored phenotypical and metabolic changes. Sedentary myopathy and wild-type mice were used as controls. Exercise increased PGC-1alpha in muscle, resulting in increased mitochondrial biogenesis, and successfully stimulated residual respiratory capacity in muscle tissue. As a consequence, ATP levels were increased in exercised mice compared with sedentary myopathy animals, which resulted in a delayed onset of the myopathy and a prolonged lifespan of the exercised mice. As an added benefit, endurance exercise induced antioxidant enzymes. The overall protective effect of endurance exercise delayed the onset of the mitochondrial myopathy and increased life expectancy in the mouse model. Thus stimulating residual oxidative phosphorylation function in the affected muscle by inducing mitochondrial biogenesis through endurance exercise might offer a valuable therapeutic intervention for mitochondrial myopathy patients. PMID- 19286572 TI - Anatomic capillarization is elevated in the medial gastrocnemius muscle of mighty mini mice. AB - House mice selectively bred for high voluntary wheel running display a mini muscle (MM) phenotype wherein mass-specific mitochondrial enzyme activities are double that of normal, but muscle mass is reduced by half. In addition, mini muscles are characterized by small muscle fibers in the superficial region of the plantaris and medial gastrocnemius muscles. To determine the structural alterations facilitating aerobic metabolism in these mini-muscles, cross-sections of the medial gastrocnemius muscle of normal (N; n = 6) and mini-muscle (MM; n = 6) mice were histo- and immunochemically labeled and analyzed for fiber size, capillarization, and fiber type. On the basis of the higher mitochondrial enzyme activities in muscles of MM mice, we hypothesized that they would have greater fiber capillarization in the medial gastrocnemius than N mice. Furthermore, we hypothesized that augmented capillarization in MM would principally be a function of the smaller fibers in the superficial aspect of this muscle. On average, MM had higher capillary-to-fiber ratio and higher capillary density. Binning fibers according to size revealed that it was primarily the normal-sized fibers of the MM that had higher capillarity. The small fibers seen in the superficial region of MM were distinct from N mice in that they had heterogeneous myofibrillar ATPase staining and patchy succinate dehydrogenase staining in the interior of the fibers. These results support the hypothesis that the MM have higher indexes of capillarity, caused primarily by greater capillary number around normally sized fibers. These alterations are consistent with the superior mass-specific aerobic function of these muscles. PMID- 19286573 TI - Artificial gravity as a countermeasure to microgravity: a pilot study examining the effects on knee extensor and plantar flexor muscle groups. AB - The goal of this project was to examine the effects of artificial gravity (AG) on skeletal muscle strength and key anabolic/catabolic markers known to regulate muscle mass. Two groups of subjects were selected for study: 1) a 21 day-bed rest (BR) group (n = 7) and 2) an AG group (n = 8), which was subjected to 21 days of 6 degrees head-down tilt bed rest plus daily 1-h exposures to AG (2.5 G at the feet). Centrifugation was produced using a short-arm centrifuge with the foot plate approximately 220 cm from the center of rotation. The torque-velocity relationships of the knee extensors and plantar flexors of the ankle were determined pre- and posttreatment. Muscle biopsy samples obtained from the vastus lateralis and soleus muscles were used for a series of gene expression analyses (mRNA abundance) of key factors implicated in the anabolic vs. catabolic state of the muscle. Post/pre torque-velocity determinations revealed greater decrements in knee extensor performance in the BR vs. AG group (P < 0.04). The plantar flexors of the AG subjects actually demonstrated a net gain in the torque velocity relationship, whereas in the BR group, the responses declined (AG vs. BR, P < 0.001). Muscle fiber cross-sectional area decreased by approximately 20% in the BR group, whereas no losses were evident in the AG group. RT-PCR analyses of muscle biopsy specimens demonstrated that markers of growth and cytoskeletal integrity were higher in the AG group, whereas catabolic markers were elevated in the BR group. Importantly, these patterns were seen in both muscles. We conclude that paradigms of AG have the potential to maintain the functional, biochemical, and structural homeostasis of skeletal muscle in the face of chronic unloading. PMID- 19286574 TI - To phase 3 or not to phase 3? PMID- 19286575 TI - Metabolic penumbra in intracerebral hemorrhage. PMID- 19286576 TI - Intracerebral hemorrhage volume predicts poor neurologic outcome in children. AB - BACKGROUND AND PURPOSE: Although intracerebral hemorrhage (ICH) volume and location are important predictors of outcome in adults, few data exist in children. METHODS: A consecutive cohort of children, including full-term newborns to those younger than 18 years of age with nontraumatic, acute ICH and head CT available for analysis were studied. Clinical information was abstracted via chart review. Hemorrhage volume was expressed as percentage of total brain volume (TBV) with large hemorrhage defined as >or=4% of TBV. Hemorrhages were manually traced on each head CT slice and volumes were calculated by multiplying by slice thickness. Location was classified as supratentorial or infratentorial. Logistic regression was used to identify predictors of poor neurological outcome, defined as a Glasgow outcome scale or=4% of TBV (P=0.03). In multivariate analysis, hemorrhage >or=4% of TBV (OR, 22.5; 95% CI, 1.4-354; P=0.03) independently predicted poor outcome 30 days after ICH. In this small sample, infratentorial hemorrhage location and the presence of intraventricular hemorrhage did not predict poor outcome. CONCLUSIONS: ICH volume predicts neurological outcome at 30 days in children, with worst outcome when hemorrhage is >or=4% of TBV. Location and ICH etiology may also be important. These findings identify children with ICH who are candidates for aggressive management and may influence counseling regarding prognosis. PMID- 19286577 TI - Postcontrast CT extravasation is associated with hematoma expansion in CTA spot negative patients. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess the effect of postcontrast CT (PCCT) leakage (PCL) on hematoma growth in CTA spot negative patients. METHODS: A retrospective study of 61 patients presenting within 6 hours of primary ICH onset imaged with CT angiography (CTA) and PCCT. Presence of CTA spot sign and PCL were documented. PCL was defined as the presence of contrast extravasation on the PCCT study at a location remote from the CTA spot sign if present. Hematoma expansion was defined as >6 mL or 30% hematoma enlargement. Patients were dichotomized by CTA spot sign presence and PCL and compared for baseline demographic data, hematoma size, and growth using the unpaired t test and Mann-Whitney test for continuous and categorical data, respectively. A probability value <0.05 was considered significant. RESULTS: PCL was present in 11/61 patients (18%), occurring in 5 without a spot sign (45%). Spot negative PCL patients demonstrated larger absolute (P=0.02) and percentage hematoma growth (P=0.02) compared to those without PCL. The mean volume and percent increase was 6.7 mL and 26%, respectively. Inclusion of PCL together with CTA spot sign as risk factor for hematoma expansion increased sensitivity from 0.78 (95% CI; 0.52 to 0.94) to 0.94 (95% CI; 0.72 to 1.00) and NPV from 0.90 (95% CI; 0.76 to 0.97) to 0.97 (95% CI; 0.85 to 1.00). CONCLUSIONS: Inclusion of PCCT in the investigation of ICH patients allows detection of PCL which, together with the CTA spot sign, increases sensitivity and negative predictive value for predicting hematoma expansion. This finding should be validated in larger studies. PMID- 19286578 TI - Inducible nitric oxide synthase promoter polymorphism affords protection against cognitive dysfunction after carotid endarterectomy. AB - BACKGROUND AND PURPOSE: Cognitive dysfunction occurs in 9% to 23% of patients during the first month after carotid endarterectomy (CEA). A 4-basepair (AAAT) tandem repeat polymorphism (either 3 or 4 repeats) has been described in the promoter region of inducible nitric oxide synthase (iNOS), a gene with complex roles in ischemic injury and preconditioning against ischemic injury. We investigated whether the 4-repeat variant (iNOS(+)) affects the incidence of cognitive dysfunction after CEA. METHODS: One-hundred eighty-five CEA and 60 spine surgery (control) subjects were included in this nested cohort analysis. Subjects underwent a battery of 7 neuropsychometric tests before and 1 day and 1 month after surgery. Multivariate logistic regression analyses were performed to determine if the iNOS promoter variant was independently associated with the incidence of cognitive dysfunction at 1 day and 1 month. Further, all right-hand dominant CEA subjects were grouped by operative side and performance on each test was compared between iNOS(+) and iNOS(-) groups. RESULTS: Forty-four of 185 CEA subjects had at least 1 iNOS promoter allele containing 4 copies of the tandem repeat (iNOS(+)). iNOS(+) status was significantly protective against moderate/severe cognitive dysfunction 1 month after CEA. Right-hand-dominant iNOS(+) CEA subjects undergoing left-side CEA performed significantly better than iNOS(-) subjects on a verbal learning test and those undergoing right-side CEA performed significantly better on a test of visuospatial function. CONCLUSIONS: We demonstrate an iNOS promoter polymorphism variant provides protection against moderate/severe cognitive dysfunction 1 month after CEA. Further, this protection appears to involve cognitive domains localized ipsilateral to the operative carotid artery. PMID- 19286579 TI - Effects of combined peripheral nerve stimulation and brain polarization on performance of a motor sequence task after chronic stroke. AB - BACKGROUND AND PURPOSE: Recent work demonstrated that application of peripheral nerve and cortical stimulation independently can induce modest improvements in motor performance in patients with stroke. The purpose of this study was to test the hypothesis that combining peripheral nerve stimulation (PNS) to the paretic hand with anodal direct current stimulation (tDCS) to the ipsilesional primary motor cortex (M1) would facilitate beneficial effects of motor training more than each intervention alone or sham (tDCS(Sham) and PNS(Sham)). METHODS: Nine chronic stroke patients completed a blinded crossover designed study. In separate sessions, we investigated the effects of single applications of PNS+tDCS, PNS+tDCS(Sham), tDCS+PNS(Sham), and PNS(Sham)+tDCS(Sham) before motor training on the ability to perform finger motor sequences with the paretic hand. RESULTS: PNS+tDCS resulted in a 41.3% improvement in the number of correct key presses relative to PNS(Sham)+tDCS(Sham), 15.4% relative to PNS+tDCS(Sham), and 22.7% relative to tDCS+PNS(Sham). These performance differences were maintained 1 and 6 days after the end of the training. CONCLUSIONS: These results indicate that combining PNS with tDCS can facilitate the beneficial effects of training on motor performance beyond levels reached with each intervention alone, a finding of relevance for the neurorehabilitation of motor impairments after stroke. PMID- 19286580 TI - Fibrinogen independently predicts the development of ischemic stroke in a Taiwanese population: CVDFACTS study. AB - BACKGROUND AND PURPOSE: Of few prospective studies that have focused on the relationship between fibrinogen and ischemic stroke (IS) in Asian populations, the findings were inconsistent with those conducted in Western countries. Therefore, we aimed to investigate the temporal relationship between fibrinogen levels (plus several related parameters) and IS in a community-based study in Taiwan. METHODS: Baseline data from 3281 adults (>or=20 years of age) in the Cardiovascular Diseases Risk Factor Two-Township Study were linked to incidental IS status derived from insurance claims and death certificate records. Hazard ratios and 95% CIs of clotting factors (fibrinogen, factor VII, factor VIII, and antithrombin-III) for IS events were estimated using Cox proportional hazard models. RESULTS: With 10.4 years (average) follow-up, 128 persons developed IS (3.75 per 1000 person-years). As expected, elevated blood pressure and diabetes were independent predictors of IS events. A dose-response relationship was found in univariate analysis between IS risk and tertiles of fibrinogen (hazard ratio, 3.73; 2.19 to 1.00), factor VII (hazard ratio, 1.86; 1.35 to 1.00), and factor VIII (2.97; 1.70 to 1.00), respectively, but not for antithrombin-III. After adjusting for confounding and known risk factors, fibrinogen independently predicted IS events. A 72% increase (hazard ratio, 1.72; 1.02 to 2.90) in IS risk was observed for individuals with fibrinogen >or=8.79 mumol/L compared with those <7.03 micromol/L. CONCLUSIONS: In addition to hypertension and diabetes, fibrinogen independently predicted future IS risk. We suggest that fibrinogen may be considered in the risk assessment model for IS in the Taiwanese population. PMID- 19286581 TI - Cost-effectiveness of patient selection using penumbral-based MRI for intravenous thrombolysis. AB - BACKGROUND AND PURPOSE: Better selection of patients for intravenous recombinant tissue plasminogen activator (IV tPA) treatment may improve clinical outcomes. We examined the cost-effectiveness of adding penumbral-based MRI to usual computed tomography (CT)-based methods to identify patients for IV tPA treatment. METHODS: A decision-analytic model estimated the lifetime costs and outcomes associated with penumbral-based MRI selection in a patient population similar to that enrolled in the IV tPA clinical trials. Inputs were obtained from published literature, clinical trial data, claims databases, and expert opinion. Outcomes included cost per life-year saved and cost per quality-adjusted life-year (QALY) gained. Costs and outcomes were discounted at 3% annually. Sensitivity analyses were conducted. RESULTS: The addition of penumbral-based MRI selection increased total cost by $103 over the patient's remaining lifetime. Penumbral-based MRI selection resulted in favorable outcomes (modified Rankin Scale or=33% or >or=12.5 mL ICH growth; and (3) radial growth >1 mm between baseline and 24-hour CT scan. Patients were divided into tertiles of baseline ICH volume: "small" (0 to 10 mL), "medium" (10 to 25 mL), and "large" (25 to 106 mL). RESULTS: Inter- and intrarater agreements for the novel scales exceeded 85% (+/-1 category). Median growth was significantly higher in the large volume group compared with the small group (P<0.001) and in heterogeneous compared with homogeneous ICH (P=0.008). Median growth trended higher in irregular ICHs compared with regular ICHs (P=0.084). Small ICHs were more regularly shaped (43%) than medium (17%) and large (3%) ICHs (P<0.001). Small ICHs were more homogeneous (73%) compared with medium (37%) and large (17%) ICHs (P<0.001). Adjusting for baseline ICH volume and time to scan, density heterogeneity, but not shape irregularity, independently predicted ICH growth (P=0.046) on a continuous growth scale. CONCLUSIONS: Large ICHs were significantly more irregular in shape, heterogeneous in density, and had greater growth. Density heterogeneity independently predicted ICH growth using some definitions. PMID- 19286591 TI - Flow-induced dilation is mediated by Akt-dependent activation of endothelial nitric oxide synthase-derived hydrogen peroxide in mouse cerebral arteries. AB - BACKGROUND AND PURPOSE: Endothelial nitric oxide synthase produces superoxide under physiological conditions leading to hydrogen peroxide (H(2)O(2)) -dependent dilations to acetylcholine in isolated mouse cerebral arteries. The purpose of this study was to investigate whether H(2)O(2) was involved in flow-mediated dilation (FMD). METHODS: Cerebral arteries were isolated from 12+/-2-week-old C57Bl/6 male mice. FMD (0 to 10 microL/min, 2-microL step increase at constant internal pressure) was induced in vessels preconstricted with phenylephrine (30 micromol/L). Simultaneously to diameter acquisition, H(2)O(2) or nitric oxide production was detected by the fluorescent dyes CMH(2)CFDA or 4,5 diaminofluorescein diacetate, respectively. Results are expressed as mean+/-SEM of 6 to 8 mice. RESULTS: FMD (at 10 microL/min, 25+/-3% of maximal diameter) was prevented (P<0.05) by endothelium removal (6+/-1%) or endothelial nitric oxide synthase inhibition with N-nitro-L-arginine (11+/-1%) but not by the specific nitric oxide scavenger 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl3-oxide (24+/-3%). Addition of PEG-catalase and silver diethyl dithio-carbamate (superoxide dismutase inhibitor) reduced (P<0.05) FMD to 10+/-2% and 15+/-1%, respectively. Simultaneously to FMD, H(2)O(2)-associated rise in fluorescence (+133+/-19 a.u.) was prevented by N-nitro-L-arginine, PEG-catalase, and silver diethyl dithio-carbamate (+55+/-10, +64+/-4, and +50+/-10 a.u., respectively; P<0.05). Inhibition of FMD by PEG-catalase was fully restored by the addition of tetrahydrobiopterin, a cofactor of endothelial nitric oxide synthase (23+/-3%); this functional reversal in dilation was associated with the simultaneous increase in nitric oxide-associated fluorescence (+418+/-58 a.u., P<0.05), which was prevented by 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl3-oxide (+93+/-26 a.u.). Akt inhibition with triciribine prevented FMD and H(2)O(2)-associated rise in fluorescence (3+/-1% and +23+/-4% a.u., respectively; P<0.05), but not acetylcholine-induced dilation. CONCLUSIONS: In healthy C57Bl/6 mouse cerebral arteries, Akt-dependent activation of endothelial nitric oxide synthase-derived H(2)O(2) mediates flow-dependent dilation. PMID- 19286593 TI - Cerebrovascular reactivity measured by near-infrared spectroscopy. AB - BACKGROUND AND PURPOSE: The pressure reactivity index (PRx) describes cerebral vessel reactivity by correlation of slow waves of intracranial pressure (ICP) and arterial blood pressure. In theory, slow changes in the relative total hemoglobin (rTHb) measured by near-infrared spectroscopy are caused by the same blood volume changes that cause slow waves of ICP. Our objective was to develop a new index of vascular reactivity, the hemoglobin volume index (HVx), which is a low-frequency correlation of arterial blood pressure and rTHb measured with near-infrared spectroscopy. METHODS: Gradual hypotension was induced in piglets while cortical laser-Doppler flux was monitored. ICP was monitored, and rTHb was measured continuously using reflectance near-infrared spectroscopy. The HVx was recorded as a moving linear correlation between slow waves (20 to 300 seconds) of arterial blood pressure and rTHb. Autoregulation curves were constructed by averaging values of the PRx or HVx in 5-mm Hg bins of cerebral perfusion pressure. RESULTS: The laser-Doppler flux-determined lower limit of autoregulation was 29.4+/-6.7 mm Hg (+/-SD). Coherence between rTHb and ICP was high at low frequencies. HVx was linearly correlated with PRx. The PRx and HVx both showed higher values below the lower limit of autoregulation and lower values above the lower limit of autoregulation. Areas under the receiver operator characteristic curves were 0.88 and 0.85 for the PRx and HVx, respectively. CONCLUSIONS: Coherence between the rTHb and ICP waveforms at the frequency of slow waves suggests that slow waves of ICP are related to blood volume changes. The HVx has potential for further development as a noninvasive alternative to the PRx. PMID- 19286594 TI - Transient focal increase in perihematomal glucose metabolism after acute human intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Progressive perihematomal cell death over 3 to 4 days has been described after experimental intracerebral hemorrhage (ICH). We investigated whether progressive perihematomal damage occurs in human subjects by measuring relative changes in regional cerebral glucose metabolism with (18)F fluorordeoxyglucose (FDG) positron emission tomography at multiple time points during the first week after ICH. METHODS: Thirteen subjects with a median hematoma volume of 22 cm(3) were studied 1.0+/-0.3, 2.9+/-0.8, and 6.7+/-1.6 days after ICH. Normalized mean counts in 5 concentric annular 2-mm-thick perihematomal volumes-of-interest (VOIs) were compared to the initial study. Next, automated searches with 0.5 to 5.0 mL spherical VOIs identified maximum focal changes in normalized counts compared to the initial study. RESULTS: No annular or focal decrease in perihematomal FDG uptake developed. Instead, FDG uptake significantly increased at session #2 in the first 3 2-mm annular VOIs (9.2%+/-14.2, 7.8%+/-11.3, 5.9%+/-9.0), returning to baseline at session #3. The VOI search identified focal regions of increased perihematomal FDG uptake relative to the contralateral control hemispheres in 6 subjects, which accounted for the annular increase. CONCLUSIONS: Perihematomal glucose metabolism increased transiently in a subset of patients 2 to 4 days after acute ICH. These transient focal increases in glucose metabolism occurring in the brain after acute ICH demonstrate that there are ongoing processes in response to injury that last for days. Although further studies are needed to elucidate their pathophysiology, these processes may be indicative of a prolonged window for intervention to improve neurological outcome. PMID- 19286595 TI - Effects of deferoxamine on intracerebral hemorrhage-induced brain injury in aged rats. AB - BACKGROUND AND PURPOSE: Deferoxamine (DFX) reduces brain edema, neuronal death, and neurological deficits after intracerebral hemorrhage (ICH) in young rats. In the present study, we investigated whether DFX is effective on brain injury after ICH in aged rats and examined dose dependency. METHODS: Male Fischer 344 rats (18 months old) had an intracaudate injection of 100 microL autologous whole blood and were treated with different doses of DFX (10, 50, and 100 mg/kg) or vehicle 2 and 6 hours post-ICH and then every 12 hours up to 7 days. Rats were euthanized at Day 3 for brain edema determination and Day 56 for brain atrophy measurement. Behavioral tests were performed during the experiments. RESULTS: All 3 doses of DFX attenuated perihematomal brain edema at 3 days (eg, at dose 50 mg/kg, 80.4+/ 0.5 versus 81.6+/-0.9% in the vehicle-treated group, P<0.01). Fifty and 100 mg/kg DFX also reduced ICH-induced ventricle enlargement, caudate atrophy, and ICH induced neurological deficits in aged rats. However, although 10 mg/kg DFX reduced ventricle enlargement and forelimb-placing deficits, it did not reduce caudate atrophy and corner turn deficits. CONCLUSIONS: These results indicate that DFX can reduce ICH-induced brain injury in aged as well as young rats and that a dose >10 mg/kg is the optimal dose of DFX in this model. PMID- 19286596 TI - Effective glycemic control with aggressive hyperglycemia management is associated with improved outcome in aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Hyperglycemia strongly predicts poor outcome in patients with aneurysmal subarachnoid hemorrhage, but the effect of hyperglycemia management on outcome is unclear. We studied the impact of glycemic control on outcome of patients with aneurysmal subarachnoid hemorrhage. METHODS: A prospective intensive care unit database was used to identify 332 patients with hyperglycemic aneurysmal subarachnoid hemorrhage admitted between January 2000 and December 2006. Patients treated with an aggressive hyperglycemia management (AHM) protocol after 2003 (N=166) were compared with 166 patients treated using a standard hyperglycemia management before 2003. Within the AHM group, outcome was compared between patients who achieved good (mean glucose burden <1.1 mmol/L) and poor (mean glucose burden >or=1.1 mmol/L) glycemic control. Poor outcome was defined as modified Rankin scale >or=4 at 3 to 6 months. Multivariable logistic regression models correcting for temporal trend were used to quantify the effect of AHM on poor outcome. RESULTS: Poor outcome in AHM-treated patients was lower (28.31% versus 40.36%) but was not statistically significant after correcting for temporal trend. However, good glycemic control significantly reduced the incidence of poor outcome (OR, 0.25; 95% CI, 0.08 to 0.80; P=0.02) compared with patients with poor glycemic control within the AHM group. No difference in the rate of clinical vasospasm or the development of delayed ischemic neurological deficit was seen before and after AHM protocol implementation. CONCLUSIONS: AHM results in good glucose control and significantly reduces the odds for poor outcome after aneurysmal subarachnoid hemorrhage in glucose-controlled patients. Further studies are needed to confirm these results. PMID- 19286597 TI - Swallowing disturbance pattern relates to brain lesion location in acute stroke patients. AB - BACKGROUND AND PURPOSE: The relationship of brain lesion location and swallowing disturbance pattern has been poorly studied in acute stroke patients. METHODS: Sixty patients with first-ever acute ischemic stroke at clearly assessed location and clinical signs of dysphagia were studied. Swallowing-related parameters rated clinically and fiberendoscopically were attention deficit, buccofacial apraxia, orofacial paresis, gag reflex, delay of pharyngeal swallow, pharyngeal contraction, larynx elevation, function of upper esophageal sphincter (UES), and aspiration severity. RESULTS: Attention deficit was independently predicted only by parietotemporal infarction, buccofacial apraxia by left-sided parietotemporal infarction, orofacial paresis by infarction encompassing upper motor neuron of cranial nerves, and impaired UES opening by lateral medullary infarction. Other swallowing parameters were not related to lesion topology. On posthoc analysis, pneumonia within 21 days after stroke was predicted only by insular lesion. CONCLUSIONS: Distinct acute brain lesion locations result in characteristic swallowing disturbance patterns. Dysphagic patients with insular stroke appear to have even higher risk of pneumonia suggesting a further associated factor promoting infection in these subjects. PMID- 19286598 TI - A method to determine stroke trial success using multidimensional pooled control functions. AB - BACKGROUND AND PURPOSE: Many early phase trials in stroke have not been subsequently confirmed. Randomization balance in baseline factors that influence outcome are difficult to achieve and may be partly responsible for misleading early results. We hypothesized that comparison with an outcome function derived from a large number of pooled control arms would mitigate these randomization problems and provide a reliable predictor for decision-making before proceeding to later phase trials. We developed such a model and added a novel feature of generation of multidimensional 95% prediction surfaces by which individual studies could be compared. We performed a proof-of-principle study with published clinical trials, determining whether our method correctly identified known outcomes. METHODS: The control arms from all randomized, controlled trials for acute stroke with >or=10 subjects, including baseline National Institute of Health Stroke Scale, age, and 3-month outcomes published between 1994 and May 2008, were identified. A Matlab program (PPREDICTS) was written to generate outcome functions based on these parameters. Published treatment trials were compared with these 95% intervals to determine whether it successfully identified positive and negative trials. RESULTS: Models of mortality and functional outcome were successfully generated (mortality: R(2)=0.69; functional outcome, modified Rankin Scale 0 to 2: R(2)=0.81; both P<0.0001). The National Institute of Neurological Diseases and Stroke intravenous recombinant tissue plasminogen activator trial and 3 studies yet to be subjected to Phase III study had modified Rankin Scale 0 to 2 outcomes above the 95% prediction interval. Sixteen treatment arm outcomes fell within prediction surface bounds. This group included 2 major trials, Stroke-Acute Ischemic NXY Treatment and Abciximab Emergent Stroke Treatment Trial, that initially appeared promising but went on to negative Phase III results. CONCLUSIONS: This proof-of-principle analysis confirmed all positive and negative clinical stroke trial results and identified some promising therapies. The use of a pooled standard treatment group function combined with statistical bounds may improve selection of early studies for further study. This method may be applicable to any condition in which baseline factors influence outcome and at any stage of the development process. PMID- 19286599 TI - Framingham stroke risk function in a large population-based cohort of elderly people: the 3C study. AB - BACKGROUND AND PURPOSE: External validation of the Framingham stroke risk function has been rarely performed. We assessed its predictive ability in a population-based cohort of French elderly. METHODS: The sample comprised 6913 subjects from the 3C Study, aged 65 to 84 at baseline, who were followed up to 6 years. Predictive accuracy of the original Framingham stroke risk function was assessed in a 3-step procedure: comparison between the Framingham and 3C cohorts of the prevalence of selected risk factors and the associated relative risks (RR) for stroke, comparison of the predicted to the observed number of stroke events (calibration), and ability to separate high-risk from low-risk participants (discrimination). We also compared predictive performances of the original Framingham, the recalibrated Framingham, and the local stroke risk functions. RESULTS: During follow-up, 110 incident strokes occurred. For most risk factors, RRs were comparable between the 2 cohorts, except for age in women. The original Framingham stroke risk function applied to the 3C cohort overestimated the 6-year absolute risk for stroke by a factor of 3.7 for men and 4.4 for women. However, the recalibrated Framingham and 3C functions did not show any over- or underestimation of stroke risk. The 3 stroke risk functions (original, recalibrated, and 3C) provided acceptable discrimination with areas under the ROC curve ranging from 0.67 to 0.73. CONCLUSIONS: The original Framingham stroke risk function strongly overestimated the stroke risk for 3C participants. Derived Framingham stroke score sheets should not be directly used by physicians in the French elderly population. PMID- 19286600 TI - Is it ethical to have a placebo arm in reperfusion trials in the 3- to 6-hour time window? Yes. PMID- 19286601 TI - Early-stage investigations of ultrasmall superparamagnetic iron oxide-induced signal change after permanent middle cerebral artery occlusion in mice. AB - BACKGROUND AND PURPOSE: MR signal changes after intravenous ultrasmall superparamagnetic iron oxide (USPIO) injection are related to inflammatory cells at the subacute stages after focal cerebral injury. However, at the early stages, the interpretation of USPIO-related MR signal alterations remains controversial. Here, we compared MR signal changes after intravenous USPIO injection with the histological iron and macrophage distribution during the first 24 hours in a rodent model of acute stroke. METHODS: Multiparametric MRI at 7T and histological USPIO distribution were confronted from 6 to 24 hours after permanent middle cerebral artery occlusion in mice. Blood-brain barrier disruption was assessed using gadolinium MRI and immunoglobulin staining. Prussian blue staining was performed to depict the USPIO brain distribution. USPIO uptake by phagocytes was assessed by immunochemistry on brain tissue, peripheral blood cells, and monocyte cells derived from bone marrow culture. RESULTS: After USPIO injection, 4 areas of early signal change were observed on every MRI. In all these areas, iron particles were mostly free whether detected in the vascular and cerebrospinal fluid compartments or in the interstitium. Within the first 24 hours, USPIO loaded cells were not detected in the blood of injured mice or in cultured monocytic cells incubated with USPIO at plasmatic concentration. CONCLUSIONS: These results suggest that, in this model, early reproducible USPIO-related MR signal changes are mainly caused by passive diffusion of free USPIO after blood brain barrier leakage and by intravascular trapping rather than by peripheral phagocyte infiltration. PMID- 19286602 TI - Blood markers for the prognosis of ischemic stroke: a systematic review. AB - BACKGROUND AND PURPOSE: The performance of validated prognostic clinical models in acute ischemic stroke might be improved by addition of data on blood biomarkers. METHODS: We searched Medline and EMBASE from 1966 to January 2007 for studies of blood markers in patients with ischemic stroke and an assessment of outcome (death, disability, or handicap). We adopted several strategies to reduce bias. RESULTS: Studies were generally small (median number of subjects, 85; interquartile range, 49 to 184). Few had evidence of a sample size calculation (7 of 82 [9%]) or reported blinding to whether patients had stroke (21 of 82 [26%]). Of the 66 studies reporting a measure of association, 10 did not adjust for age or stroke severity, 14 adjusted for age, 7 adjusted for severity, and 35 adjusted for both; 30% (20 of 66) used a data-dependent threshold to predict good or bad outcome. There was evidence of within-study reporting bias and publication bias. Cardiac markers showed the most consistent association with poor outcome. CONCLUSIONS: Blood biomarkers might provide useful information to improve the prediction of outcome after acute ischemic stroke. However, this review showed that many studies were subject to bias. Although some markers had some predictive ability, none of the studies was able to demonstrate that the biomarker added predictive power to a validated clinical model. The clinical usefulness of blood biomarkers for predicting prognosis in the setting of ischemic stroke has yet to be established. PMID- 19286603 TI - Long-term recurrent subarachnoid hemorrhage after adequate coiling versus clipping of ruptured intracranial aneurysms. AB - BACKGROUND AND PURPOSE: Coiling is increasingly used as treatment for intracranial aneurysms. Despite its favorable short-term outcome, concerns exist about long-term reopening and inherent risk of recurrent subarachnoid hemorrhage (SAH). We hypothesized a higher risk for recurrent SAH after adequate coiling compared with clipping. METHODS: Patients with ruptured intracranial aneurysms coiled between 1994 and 2002 with adequate (>90%) aneurysm occlusion at 6-month follow-up angiograms were included. We interviewed these patients about new episodes of SAH. By survival analysis, we assessed the cumulative incidence of recurrent SAH after coiling and compared it with the incidence of recurrent SAH in a cohort of 748 patients with clipped aneurysms by calculating age and sex adjusted hazard ratios. RESULTS: Of 283 coiled patients with a total follow-up of 1778 patient-years (mean, 6.3 years), one patient had a recurrent SAH (0.4%) and 2 patients had a possible recurrent SAH. For recurrent SAH within the first 8 years after treatment, the cumulative incidence was 0.4% (95% CI, -0.4 to 1.2) after coiling versus 2.6% (95% CI, 1.2 to 4.0) after clipping (hazard ratio, 0.2; 95% CI, 0.03 to 1.6). For possible and confirmed recurrent SAH combined, the cumulative incidence was 0.7% (95% CI, 0.3 to 1.7) after coiling versus 3.0% (95% CI, 1.3 to 4.6) after clipping (hazard ratio, 0.7; 95% CI, 0.2 to 2.3). CONCLUSIONS: Patients with adequately occluded aneurysms by coiling at short-term follow-up are at low risk for recurrent SAH in the long term. Within the first 8 years after treatment, the risk of recurrent SAH is not higher after adequate coiling than after clipping. PMID- 19286604 TI - Periventricular white matter lucencies relate to low vitamin B12 levels in patients with small vessel stroke. AB - BACKGROUND AND PURPOSE: Blood-brain barrier dysfunction may be an early phenomenon in the development of the small vessel disease, which underlies white matter lesions. Because vitamin B12 plays a role in maintaining the integrity of the blood-brain barrier, we studied serum vitamin B12 level in relation to such lesions. METHODS: In 124 patients with first lacunar stroke, we measured serum vitamin B12 level and rated the degree of white matter lesions on MRI. RESULTS: Mean vitamin B12 level was 202 pmol/L (SD, 68.9). Thirty-nine patients (31.5%) had a vitamin B12 level less than the lower reference value of 150 pmol/L. Lower vitamin B12 level was (statistically significant) associated with more severe periventricular white matter lesions (odds ratio/100 pmol/L decrease, 1.773; 95% CI, 1.001-3.003), but not with deep white matter lesions (odds ratio/100 pmol/L decrease, 1.441; 95% CI, 0.881-2.358; ordered multivariate regression analysis). CONCLUSIONS: More severe periventricular white matter lesions in lacunar stroke patients relate to lower vitamin B12 levels. A possible causal relationship should now be studied prospectively. PMID- 19286605 TI - The ethics of thrombolytic trials beyond 3 (or 4.5) hours: randomized controlled trials are required to change clinical practice. PMID- 19286606 TI - Is it ethical to have a placebo arm in reperfusion trials in the 3- to 6-hour time window? No: time frame or time gain? PMID- 19286607 TI - Akt-mediated transactivation of the S1P1 receptor in caveolin-enriched microdomains regulates endothelial barrier enhancement by oxidized phospholipids. AB - Endothelial cell (EC) barrier dysfunction results in increased vascular permeability, leading to increased mass transport across the vessel wall and leukocyte extravasation, the key mechanisms in pathogenesis of tissue inflammation and edema. We have previously demonstrated that OxPAPC (oxidized 1 palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine) significantly enhances vascular endothelial barrier properties in vitro and in vivo and attenuates endothelial hyperpermeability induced by inflammatory and edemagenic agents via Rac and Cdc42 GTPase dependent mechanisms. These findings suggested potential important therapeutic value of barrier-protective oxidized phospholipids. In this study, we examined involvement of signaling complexes associated with caveolin enriched microdomains (CEMs) in barrier-protective responses of human pulmonary ECs to OxPAPC. Immunoblotting from OxPAPC-treated ECs revealed OxPAPC-mediated rapid recruitment (5 minutes) to CEMs of the sphingosine 1-phosphate receptor (S1P(1)), the serine/threonine kinase Akt, and the Rac1 guanine nucleotide exchange factor Tiam1 and phosphorylation of caveolin-1, indicative of signaling activation in CEMs. Abolishing CEM formation (methyl-beta-cyclodextrin) blocked OxPAPC-mediated Rac1 activation, cytoskeletal reorganization, and EC barrier enhancement. Silencing (small interfering RNA) Akt expression blocked OxPAPC mediated S1P(1) activation (threonine phosphorylation), whereas silencing S1P(1) receptor expression blocked OxPAPC-mediated Tiam1 recruitment to CEMs, Rac1 activation, and EC barrier enhancement. To confirm our in vitro results in an in vivo murine model of acute lung injury with pulmonary vascular hyperpermeability, we observed that selective lung silencing of caveolin-1 or S1P(1) receptor expression blocked OxPAPC-mediated protection from ventilator-induced lung injury. Taken together, these results suggest Akt-dependent transactivation of S1P(1) within CEMs is important for OxPAPC-mediated cortical actin rearrangement and EC barrier protection. PMID- 19286608 TI - The subendothelial extracellular matrix modulates JNK activation by flow. AB - Atherosclerosis begins as local inflammation of artery walls at sites of disturbed flow. JNK (c-Jun NH(2)-terminal kinase) is thought to be among the major regulators of flow-dependent inflammatory gene expression in endothelial cells in atherosclerosis. We now show that JNK activation by both onset of laminar flow and long-term oscillatory flow is matrix-specific, with enhanced activation on fibronectin compared to basement membrane protein or collagen. Flow induced JNK activation on fibronectin requires new integrin ligation and requires both the mitogen-activated protein kinase kinase MKK4 and p21-activated kinase. In vivo, JNK activation at sites of early atherogenesis correlates with the deposition of fibronectin. Inhibiting p21-activated kinase reduces JNK activation in atheroprone regions of the vasculature in vivo. These results identify JNK as a matrix-specific, flow-activated inflammatory event. Together with other studies, these data elucidate a network of matrix-specific pathways that determine inflammatory events in response to fluid shear stress. PMID- 19286609 TI - In Memoriam: B. Lowell Langille (1947-2008). PMID- 19286610 TI - Tiny bubbles and endocytosis? PMID- 19286611 TI - Intracellular signaling of LOX-1 in endothelial cell apoptosis. PMID- 19286612 TI - Maternal-fetal cholesterol transport in the placenta: good, bad, and target for modulation. PMID- 19286613 TI - Arterial-venous specification during development. AB - The major arteries and veins of the vertebrate circulatory system are formed early in embryonic development, before the onset of circulation, following de novo aggregation of "angioblast" progenitors in a process called vasculogenesis. Initial embryonic determination of artery or vein identity is regulated by variety of genetic factors that work in concert to specify endothelial cell fate, giving rise to 2 distinct components of the circulatory loop possessing unique structural characteristics. Work in multiple in vivo animal model systems has led to a detailed examination of the interacting partners that determine arterial and venous specification. We discuss the hierarchical arrangement of many signaling molecules, including Hedgehog (Hh), vascular endothelial growth factor (VEGF), Notch, and chicken ovalbumin upstream-transcription factor II (COUP-TFII) that promote or inhibit divergent pathways of endothelial cell fate. Elucidation of the functional role of these genetic determinants of blood vessel specification together with the epigenetic factors involved in subsequent modification of arterial-venous identity will allow for potential new therapeutic targets for vascular disorders. PMID- 19286614 TI - Painful swallowing after herpes stomatitis. PMID- 19286615 TI - Extreme hyponatremia in a child with vegetative state and water intoxication. PMID- 19286616 TI - A four-year-old with Streptococcus pneumoniae--associated hemolytic uremic syndrome and gall bladder disease necessitating cholecystectomy. PMID- 19286617 TI - Ludwig's angina in the pediatric population. AB - As many as 1 in 3 of cases of Ludwig's angina occur in children and adolescents, and pediatricians are therefore ideally situated to detect these individuals at an early stage of their potentially life-threatening disease. The early identification and referral of children afflicted with Ludwig's angina to tertiary care centers allows for the rapid initiation of medical therapy and the consultation of those emergency services critical to providing such patients with optimal diagnostic and therapeutic interventions. This review provides an overview of the anatomical and pathophysiological considerations in Ludwig's angina and describes practical management principles to assist pediatricians in the diagnosis and treatment of this disease. Included in this review is an evidence-based algorithm for airway management. PMID- 19286618 TI - Formulary coverage for lipid-lowering drugs recommended for children. AB - OBJECTIVE/METHODS: A cross-sectional assessment to describe availability, coverage, and pediatric labeling status of lipid-lowering drugs offered by 1 private and 1 public insurance plan formularies. RESULTS: Both insurance plans had equal medication availability of bile acid sequestrants (7), statins (10), and cholesterol-absorption blockers (CAB; 1). The private plan had 3 bile acid sequestrants and 3 statins listed as preferred drugs; the CAB was not preferred. In contrast, the public plan had 5 bile acid sequestrants, 7 statins, and the CAB as preferred drugs. For medications with pediatric labeling, the private plan covered 50% as preferred drugs whereas the public plan covered 70% as preferred drugs. CONCLUSIONS: If new recommendations of the American Academy of Pediatrics for treatment of dyslipidemia in children were implemented today, children with the public plan would have equal choice but better coverage of lipid-lowering drugs as preferred drugs, including those with FDA approval, compared with children with the private plan. PMID- 19286619 TI - Purulent vaginal discharge secondary to ureteral ectopia. PMID- 19286620 TI - Successful treatment with high-dose intravenous immunoglobulin for parvovirus b19 infection associated with acute fulminant hepatitis in a chinese child. PMID- 19286621 TI - Building communication between professionals at children's specialty hospitals and the medical home. AB - OBJECTIVE: Build a quality improvement (QI) intervention to improve communication between a children's specialty hospital and referring primary care providers (PCPs). METHODS: A network of charitable children's hospitals identified improving communication as a systemwide goal. At one model hospital, we used qualitative telephone interviewing of hospital specialists and staff, and referring PCPs, to characterize the communication system and identify potential improvements. We identified potential high-impact areas through content analysis and developed a QI change package with hospital leadership. RESULTS: Participants described inconsistent communication, with no systematic identification of PCPs. Families were the typical means of inter-physician communication. Multiple non PCP referral sources were a major contributor to communication breakdowns. Respondents identified a system for identification and communication with PCPs as an essential first step. CONCLUSIONS: Systems for communication with PCPs are underdeveloped at a children's charitable specialty hospital. Straightforward changes could build an effective system that is generalizable to other hospitals. PMID- 19286622 TI - An infant with diffuse lung masses. AB - Chronic granulomatous disease presenting as invasive pulmonary aspergillosis in an infant is described in this report. Symptoms of chronic, intermittent respiratory distress had been previously attributed to gastroesophageal regurgitation and asthma. The isolation of an unusual pathogen even from a nonsterile site, particularly when symptoms persist, should not be considered as a contamination but prompt further investigation to exclude phagocytic or other immune deficiencies. PMID- 19286623 TI - Another duel in the sun: weighing the balances between sun protection, tanning beds, and malignant melanoma. AB - OBJECTIVE: The purpose of this report is to put the dueling factors of risk and prevention for melanoma in perspective for the thoughtful pediatric specialist to facilitate preteen preventive health counseling. STUDY DESIGN: We examined the rate of malignant melanoma among Kentucky residents and compared this rate with indicators of tanning bed prevalence in a large metropolitan area and sunscreen sales from a major distributor. We obtained malignant melanoma annual incidence data from the Kentucky Cancer Registry, which recorded Kentucky population incidence rates over the years 1995 to 2004. The rates reflected 2 malignant melanoma classifications: pre-invasive cancer only, or both invasive and noninvasive cancers combined. RESULTS: The age-adjusted incidence rate per hundred thousand for combined invasive and pre-invasive malignant melanoma swelled from 21.9 in 1995 to 31.3 in 2004. The respective invasive-only malignant melanoma incidence rates increased less dramatically, from 17.3 to 20.7, during this same 10-year time period. Since 1983, the number of separate tanning bed businesses increased from 1 in 1983 to 119 by the mid-1990s, and then declined to about 74 separate businesses by 2003. Sunscreen sales data is uneven between states and is currently inconclusive. CONCLUSIONS: Although current data cannot draw a precise link between melanoma and the use of tanning beds, the associated risk is implicit, as the ultraviolet A (UVA) and ultraviolet B (UVB) radiation in tanning bed usage is a well-established melanoma risk factor. In advising patients, the pediatric specialist should consider that melanoma rates are poised as a balance of some known risk factors and a few potential preventive factors. PMID- 19286624 TI - The beta-agonist saga and its clinical relevance: on and on it goes. AB - The decision by the Food and Drug Administration to issue a "black box" warning for long-acting beta-agonists has been followed by a series of pharmacoepidemiological studies focusing on the safety of these drugs. However, these provide the clinician with mixed messages and do not offer clear guidance as to whether adverse responses to beta-agonists are a relevant consideration in individual patients. Simultaneously, there is a growing body of evidence that continuous or high dose-beta-agonist exposure is proinflammatory and that, paradoxically, airway hyperresponsiveness is enhanced, not attenuated. Also, pharmacological theory regarding the pathophysiological function of the beta adrenoceptor is having to be revised. A recent clinical study has even suggested that beta-blockers rather than beta-agonists may be beneficial in asthma. In practice, there are individuals in whom excessive beta-agonist use contributes adversely to poor asthma control. The recommendation that concomitant use of inhaled steroids will obviate any risks associated with beta-agonists is not in fact fool-proof. Clinicians need to be aware of how to identify and manage patients for whom beta-agonist treatment is a problem rather than a solution. They constitute a small but important subgroup of patients with difficult asthma. PMID- 19286625 TI - Repeated aerosol delivery of carboxyl-terminal modulator protein suppresses tumor in the lungs of K-rasLA1 mice. AB - RATIONALE: Difficulties in achieving long-term survival of patients with lung cancer treated with conventional therapies suggest that novel approaches are required. Recent advances in aerosol-mediated gene delivery have provided the possibility of an alternative for the safe and effective treatment of lung cancer. OBJECTIVES: To investigate the repeated effect of carboxyl-terminal modulator protein (CTMP) on multistage lung tumorigenesis. In this study, we addressed this question by studying the effects of lentivirus-based CTMP in the lungs of 9- and 13-week-old K-ras(LA1) mice, a model of lung cancer. METHODS: An aerosol of lentivirus-based CTMP was delivered into 9- and 13-week-old K-ras(LA1) mice, a model of lung cancer, through a nose-only inhalation system twice a week for 4 weeks. The effects of CTMP on lung cancer progression and Akt-related signals were evaluated. MEASUREMENTS AND MAIN RESULTS: Long-term repeated delivery of CTMP effectively reduced tumor progression in the lungs at different stages of development. Lentiviral-CTMP inhibited protein synthesis and cell cycle and altered Akt signaling pathway in the lungs of 9-week-old K-ras(LA1) mice, and increased apoptosis was observed in the lungs of 13-week-old K-ras(LA1) mice. CONCLUSIONS: Long-term repeated viral delivery of CTMP may provide a useful tool for designing lung tumor treatment. PMID- 19286626 TI - Exploring the association between severe respiratory syncytial virus infection and asthma: a registry-based twin study. AB - RATIONALE: Severe respiratory syncytial virus (RSV) infection is associated with asthma but the nature of this association is imperfectly understood. OBJECTIVES: To examine the nature of the association between severe RSV infection and asthma in a population-based sample of twins. METHODS: Data on hospitalization due to RSV infection was gathered for all twins born in Denmark between 1994 and 2000 (8,280 pairs) and linked to information on asthma obtained from hospital discharge registries and parent-completed questionnaires. Genetic variance components models and direction of causation models were fitted to the observed data. MEASUREMENTS AND MAIN RESULTS: RSV hospitalization and asthma were positively associated (r = 0.43), and genetic determinants for the two disorders overlapped completely. Modeling the direction of causation between RSV hospitalization and asthma showed that a model in which asthma "causes" RSV hospitalization fitted the data significantly better (P = 0.39 for deterioration in model fit) than a model in which RSV hospitalization "causes" asthma (P < 0.001 for deterioration in model fit), even when sex, birth weight, and maternal smoking during pregnancy were accounted for. CONCLUSIONS: RSV infection that is severe enough to warrant hospitalization does not cause asthma but is an indicator of the genetic predisposition to asthma. PMID- 19286627 TI - Baroreflex gain in children with obstructive sleep apnea. AB - RATIONALE: We previously demonstrated that children with obstructive sleep apnea have increased blood pressure associated with changes in left ventricular mass index. Others have shown in adults that blood pressure variability is an important predictor of changes in left ventricular mass. The baroreflex system buffers blood pressure changes by varying heart rate. We have thus hypothesized that (1) baroreflex system gain is increased during sleep, improving blood pressure buffering; (2) children with obstructive sleep apnea lack this baroreflex gain increase; and (3) reduced blood pressure buffering results in exaggerated blood pressure variability that is associated with end-organ damage. OBJECTIVES: Compare measures of left ventricular mass index and nighttime baroreflex gain of healthy children to those of children with obstructive sleep apnea. METHODS: A total of 169 children (50 control subjects, 63 with mild obstructive sleep apnea, and 56 with severe obstructive sleep apnea) with a mean age of 9.9 years (+/-2.2) underwent echocardiography followed by polysomnography with continuous blood pressure measurement. Baroreflex gain was calculated in time and frequency domains. MEASUREMENTS AND MAIN RESULTS: Healthy children demonstrated a nighttime pattern of increasing baroreflex gain. Children with obstructive sleep apnea had decreased nighttime baroreflex gain compared with control subjects. Nighttime blood pressure and blood pressure variability were significantly correlated with left ventricular mass index. CONCLUSIONS: Obstructive sleep apnea is associated with a decrease in nighttime baroreflex gain and an increase in blood pressure variability. This increase is correlated with changes in left ventricular mass index. PMID- 19286628 TI - Alveolar extracellular 20S proteasome in patients with acute respiratory distress syndrome. AB - RATIONALE: Repair mechanisms resulting in alveolar protein degradation in acute respiratory distress syndrome (ARDS) are largely unknown. OBJECTIVES: To test whether the 20S proteasome is present and functional in the alveolar space in patients with ARDS. METHODS: Proteasome antigenic concentration in bronchoalveolar lavage (BAL) supernatants was measured by ELISA in patients with ARDS (n = 64), acute lung injury (ALI) (n = 8), sarcoidosis (n = 13), and in healthy subjects (n = 8). Cleavage of specific fluorogenic substrates (+/ epoxomicin), I(125) albumin degradation rate, and gel filtration were used to quantify and characterize proteasomal activity. The presence of proteasomes was confirmed independently by electron microscopic techniques. MEASUREMENTS AND MAIN RESULTS: Proteasome concentrations in patients with ARDS were markedly increased (1,069 +/- 1,194 ng/ml) in comparison to healthy subjects (60.8 +/- 49.8; P < 0.001), ALI (154 +/- 43; P = 0.006), and sarcoidosis (97.6 +/- 42.2; P = 0.037). All fluorogenic substrates were hydrolyzed (Suc-LLVY-AMC, 3.6 +/- 8.8 pkat/mg; BZ VGR-AMC, 1.8 +/- 3.1; Suc-LLE-AMC, 1 +/- 1.7) by BAL supernatants of patients with ARDS, with inhibition by epoxomicin (P = 0.0001), and the majority of proteolytic activity was detected in BAL supernatant. Maximum hydrolyzing activity occurred at 660 kD and 20S proteasome was seen microscopically after purification and being released by pneumocytes type II. Proteasomal activity and albumin degradation rate in patients with ARDS were approximately 17-fold lower than in healthy subjects. Proteasomal activity in normal BAL was inhibited by BAL aliquots from patients with ARDS but not by denatured BAL, and returned to normal by purification. CONCLUSIONS: For the first time, we identified extracellular, biologically active 20S proteasome in the alveolar space of patients with ARDS in concentrations much higher than in normal subjects or in those with ALI. PMID- 19286629 TI - An expanded seed sequence definition accounts for full regulation of the hid 3' UTR by bantam miRNA. AB - MicroRNAs (miRNAs) are an abundant class of approximately 22 nucleotide (nt) long noncoding RNAs that negatively regulate gene expression post-transcriptionally through imperfect base-pairing interactions with sequences in the target messenger RNA (mRNA). We examined the interactions of the bantam miRNA with the 3' untranslated region (UTR) of the hid mRNA, and a synthetic derivative, in Drosophila S2 cells in order to define the relative contributions of proposed bantam binding sites. The contribution of the bantam miRNA to repression of reporter constructs carrying different 3' UTRs was evaluated by measuring derepression of reporter expression following the transfection of bantam complementary oligoribonucleotides (anti-bantam). Systematic excision of bantam miRNA target sequences in the hid 3' UTR identified by commonly used miRNA target prediction programs failed to relieve repression to the extent predicted by the anti-bantam experiment. However, removal of additional bantam complementary sequences (with a "seed" match to nucleotide 3-9) derepressed the reporter constructs to the full extent, arguing for a less narrow definition of the seed sequence. Further support for the potential contribution of the 3-9 seed register to microRNA-mediated gene regulation is provided by the experimental validation of several novel bantam targets identified with a more relaxed search algorithm. PMID- 19286631 TI - TIP47, a lipid cargo protein involved in macrophage triglyceride metabolism. AB - OBJECTIVE: Uptake of lipids by macrophages (MPhi) leads to lipid droplet accumulation and foam cell formation. The PAT family proteins are implicated in lipid droplet formation, but the precise function of the 47-kDa tail interacting protein (TIP47), a member of this family, is poorly defined. The present study was performed to determine the function of TIP47 in MPhi lipid metabolism. METHODS AND RESULTS: Freeze-fracture cytochemistry demonstrates that TIP47 is present in the plasma membrane of MPhi and is aggregated into clusters when the cells are incubated with oleate. Suppression of adipophilin levels using siRNA knockdown leads to migration of TIP47 from a cytoplasmic pool to the lipid droplet. Further, reduction of TIP47 decreases triglyceride levels, whereas raising TIP47 levels by expression of EGFP-TIP47 shows the opposite effect. CONCLUSION: Our results show that the TIP47 protein levels directly correlate with triglyceride levels. We propose that TIP47 may act as a carrier protein for free fatty acids and in this way participates in conversion of MPhi into foam cells. PMID- 19286630 TI - Apolipoprotein A-I and its role in lymphocyte cholesterol homeostasis and autoimmunity. AB - OBJECTIVE: The purpose of this study was to determine the effects of an atherogenic diet on immune function in LDLr(-/-), ApoA-I(-/-) mice. METHODS AND RESULTS: When LDLr(-/-), ApoA-I(-/-) (DKO), and LDLr(-/-) (SKO) mice were fed an atherogenic diet, DKO had larger peripheral lymph nodes (LNs) and spleens compared to SKO mice. LNs were enriched in cholesterol and contain expanded populations of T, B, dendritic cells, and macrophages. Expansion of all classes of LN cells was accompanied by a approximately 1.5-fold increase in T cell proliferation and activation. Plasma antibodies to dsDNA, beta2-glycoprotein I, and oxidized LDL were increased in DKO, similar to levels in diet-fed Fas(lpr/lpr) mice, suggesting the development of an autoimmune phenotype. Both LN enlargement and cellular cholesterol expansion were "prevented" when diet-fed DKO mice were treated with helper dependent adenovirus expressing apoA-I. Independent of the amount of dietary cholesterol, DKO mice consistently showed lower plasma cholesterol than SKO mice, yet greater aortic cholesterol deposition and inflammation. CONCLUSIONS: ApoA-I prevented cholesterol-associated lymphocyte activation and proliferation in peripheral LN of diet-fed DKO mice. A approximately 1.5-fold increase in T cell activation and proliferation was associated with a approximately 3-fold increase in concentrations of circulating autoantibodies and approximately 2-fold increase in the severity of atherosclerosis suggesting a common link between plasma apoA-I, inflammation, and atherosclerosis. PMID- 19286632 TI - Decision analysis supports the paradigm that indiscriminate supplementation of vitamin E does more harm than good. AB - OBJECTIVE: For many years, the prevailing concept was that LDL oxidation plays a central role in atherogenesis. As a consequence, supplementation of antioxidants, particularly vitamin E, became very popular. Unfortunately, however, the major randomized clinical trials have yielded disappointing results on the effects of vitamin E on both mortality and morbidity. Moreover, recent meta-analyses have concluded that vitamin E supplementation increases mortality. This conclusion has raised much criticism, most of it relating to three issues: (1) the choice of clinical trials to be included in the meta-analyses; (2) the end point of these meta-analyses (only mortality); and (3) the heterogeneity of the analyzed clinical trials with respect to both population and treatment. Our goal was to bring this controversy to an end by using a Markov-model approach, which is free of most of the limitations involved in using meta-analyses. METHODS AND RESULTS: We used a Markov model to compare the vitamin E supplemented virtual cohorts with nonsupplemented cohorts derived from published randomized clinical trials that were included in at least one of the major meta-analyses. The difference between the virtual supplemented and nonsupplemented cohorts is given in terms of a composite end point denoted quality-adjusted life year (QALY). The vitamin E supplemented virtual cohort had 0.30 QALY (95%CI 0.21 to 0.39) less than the nontreated virtual cohort. CONCLUSIONS: Our study demonstrates that in terms of QALY, indiscriminate supplementation of high doses of vitamin E is not beneficial in preventing CVD. Selective supplementation of vitamin E to individuals under oxidative stress requires further investigation. PMID- 19286633 TI - Role of the low-affinity leukotriene B4 receptor BLT2 in VEGF-induced angiogenesis. AB - OBJECTIVE: The leukotriene B(4) (LTB(4)) receptor BLT2 is expressed in endothelium, but no clear physiological function for it has yet been identified, especially in vascular angiogenesis. The purpose of this study is to characterize the potential function of BLT2 in vascular endothelial growth factor (VEGF) induced angiogenesis. METHODS AND RESULTS: VEGF significantly upregulates BLT2 expression in human umbilical vein endothelial cells (HUVECs), and BLT2 knockdown by siRNA or inhibition of BLT2 by a specific BLT2 antagonist LY255283 attenuates VEGF-induced angiogenesis, which was determined by its effect on the formation of tube-like structures and on transmigration. The role of BLT2 in VEGF-induced angiogenesis was more evident in vivo, where BLT2 inhibition by LY255283 almost completely blocked VEGF-induced vessel formation in Matrigel-plug assays. In addition, we found that VEGF upregulates synthesis of the BLT2 ligand, 12(S) hydroxyeicosatetraenoic acid (HETE). siRNA knockdown of 12-lipoxygenase (12-LO) expression attenuates VEGF-induced angiogenesis in HUVECs, and the addition of 12(S)-HETE to the 12-LO knockdown-HUVECs restores VEGF-induced angiogenesis. The activation of BLT2 itself by either 12(S)-HETE or LTB(4) evoked significant angiogenic phenotypes, both in vitro and in vivo. CONCLUSIONS: Our findings indicate that BLT2 plays an essential role in mediating VEGF-induced angiogenesis. PMID- 19286634 TI - Protective role of SIRT1 in diabetic vascular dysfunction. AB - OBJECTIVE: Calorie restriction (CR) prolongs the lifespan of various species, ranging from yeasts to mice. In yeast, CR extends the lifespan by increasing the activity of silencing information regulator 2 (Sir2), an NAD(+)-dependent deacetylase. SIRT1, a mammalian homolog of Sir2, has been reported to downregulate p53 activity and thereby prolong the lifespan of cells. Although recent evidence suggests a link between SIRT1 activity and metabolic homeostasis during CR, its pathological role in human disease is not yet fully understood. METHODS AND RESULTS: Treatment of human endothelial cells with high glucose decreases SIRT1 expression and thus activates p53 by increasing its acetylation. This in turn accelerates endothelial senescence and induces functional abnormalities. Introduction of SIRT1 or disruption of p53 inhibits high glucose induced endothelial senescence and dysfunction. Likewise, activation of Sirt1 prevents the hyperglycemia-induced vascular cell senescence and thereby protects against vascular dysfunction in mice with diabetes. CONCLUSIONS: These findings represent a novel mechanism of vascular cell senescence induced by hyperglycemia and suggest a protective role of SIRT1 in the pathogenesis of diabetic vasculopathy. PMID- 19286636 TI - Effects of aspirin on clot structure and fibrinolysis using a novel in vitro cellular system. AB - OBJECTIVES: The purpose of this study was to investigate the direct effects of aspirin on fibrin structure/function. METHODS AND RESULTS: Chinese Hamster Ovary cell lines stably transfected with fibrinogen were grown in the absence (0) and presence of increasing concentrations of aspirin. Fibrinogen was purified from the media using affinity chromatography, and clots were made from recombinant protein. Mean final turbidity [OD(+/-SEM)] was 0.083(+/-0.03), 0.093(+/-0.002), 0.101(+/-0.005), and 0.125(+/-0.003) in clots made from 0, 1, 10, and 100 mg/L aspirin-treated fibrinogen, respectively (P<0.05). Permeability coefficient (Ks cm2 x 10(-8)) was 1.68(+/-0.29) and 4.13(+/-0.33) comparing fibrinogen produced from cells grown with 0 mg/L and 100 mg/L aspirin respectively (P<0.05). Scanning electron microscopy confirmed a looser clot structure and increased fiber thickness of clots made from aspirin-treated fibrinogen, whereas rheometer studies showed a significant 30% reduction in clot rigidity. Fibrinolysis was quicker in clots made from aspirin-treated fibrinogen. Ex vivo studies in 3 normal volunteers given 150 mg aspirin daily for 1 week demonstrated similar changes in clot structure/function. CONCLUSION: Aspirin directly altered clot structure resulting in the formation of clots with thicker fibers and bigger pores, which are easier to lyse. This study clearly demonstrates an alternative mode of action for aspirin, which should be considered in studies evaluating the biochemical efficacy of this agent. PMID- 19286635 TI - Sphingomyelin synthase 2 is one of the determinants for plasma and liver sphingomyelin levels in mice. AB - BACKGROUND: It has been proposed that plasma sphingomyelin (SM) plays a very important role in plasma lipoprotein metabolism and atherosclerosis. Sphingomyelin synthase (SMS) is the last enzyme for SM de novo biosynthesis. Two SMS genes, SMS1 and SMS2, have been cloned and characterized. METHODS AND RESULTS: To evaluate the in vivo role of SMS2 in SM metabolism, we prepared SMS2 knockout (KO) and SMS2 liver-specific transgenic (LTg) mice and studied their plasma SM and lipoprotein metabolism. On a chow diet, SMS2 KO mice showed a significant decrease in plasma SM levels (25%, P<0.05), but no significant changes in total cholesterol, total phospholipids, or triglyceride, compared with wild-type (WT) littermates. On a high-fat diet, SMS2 KO mice showed a decrease in plasma SM levels (28%, P<0.01), whereas SMS2LTg mice showed a significant increase in those levels (29%, P<0.05), but no significant changes in other lipids, compared with WT littermates. Atherogenic lipoproteins from SMS2LTg mice displayed a significantly stronger tendency toward aggregation after mammalian sphingomyelinase treatment, compared with controls. Moreover, SMS2 deficiency significantly increased plasma apoE levels (2.0-fold, P<0.001), whereas liver specific SMS2 overexpression significantly decreased those levels (1.8-fold, P<0.01). Finally, SMS2 KO mouse plasma promoted cholesterol efflux from macrophages, whereas SMS2LTg mouse plasma prevented it. CONCLUSIONS: We therefore believe that regulation of liver SMS2 activity could become a promising treatment for atherosclerosis. PMID- 19286637 TI - Editorial comment: Montani versus Osborn exchange of views. PMID- 19286638 TI - Understanding the contribution of Guyton's large circulatory model to long-term control of arterial pressure. AB - With the publication in 1972 of a large computer model of circulatory control, Guyton and colleagues challenged the then prevailing views on how blood pressure and cardiac output were controlled. At that time, it was widely accepted that the heart controlled cardiac output and that peripheral resistance controlled arterial blood pressure. By incorporating the empirically demonstrated concepts of blood flow autoregulation and the pressure-natriuresis relationship into their mathematical model, Guyton and colleagues were able to develop a number of revolutionary concepts. Guyton's circulatory model was particularly instrumental in exploring the linkage between blood pressure and sodium balance and in demonstrating an overriding importance of renal salt and water balance in setting the long-term blood pressure level. In both the model and experimental data, any long-lasting imbalance between salt intake and salt excretion leads to a progressive alteration of the degree of filling of the vascular system and thus to parallel changes in blood pressure. In turn, changes in blood pressure alter sodium excretion, opposing the initial salt imbalance. Although Guyton's model does not include the most recent cardiovascular discoveries, the concepts underlying the basic functioning of the cardiovascular system can serve as a well built basis for the development of new, large and integrative cardiovascular models. PMID- 19286639 TI - Commentary on 'Understanding the contribution of Guyton's large circulatory model to long-term control of arterial pressure'. PMID- 19286640 TI - Current computational models do not reveal the importance of the nervous system in long-term control of arterial pressure. AB - Arterial pressure is regulated over long periods of time by neural, hormonal and local control mechanisms, which ultimately determine the total blood volume and how it is distributed between the various vascular compartments of the circulation. A full understanding of the complex interplay of these mechanisms can be greatly facilitated by the use of mathematical models. In 1967, Guyton and Coleman published a model for long-term control of arterial pressure that focused on renal control of body sodium and water and thus total blood volume. The central point of their model is that the long-term level of arterial pressure is determined exclusively by the 'renal function curve', which relates arterial pressure to urinary excretion of salt and water. The contribution of the sympathetic nervous system to setting the long-term level of arterial pressure in the model is limited. In light of the overwhelming evidence for a major role of the sympathetic nervous system in long-term control of arterial pressure and the pathogenesis of hypertension, new mathematical models for long-term control of arterial pressure may be necessary. Despite the prominence and general acceptance of the Guyton-Coleman model in the field of hypertension research, we argue here that it overestimates the importance of renal control of body fluids and total blood volume in blood pressure regulation. Furthermore, we suggest that it is possible to construct an alternative model in which sympathetic nervous system activity plays an important role in long-term control of arterial pressure independent of its effects on total blood volume. PMID- 19286641 TI - Commentary on 'Current computational models do not reveal the importance of the nervous system in long-term control of arterial pressure'. PMID- 19286642 TI - Inspirational lessons from watching swallows. PMID- 19286643 TI - Text comprehension products and processes in young, young-old, and old-old adults. AB - Aging may be associated with an increase in generalized text processing, particularly in adults older than 75 years. The current study examined text comprehension in young, young-old, and old-old adults. Experiment 1 included a comprehension measure (product) and Experiment 2 examined inferences generated during reading (process). Comprehension scores were lowest in old-old adults. Generalized and elaborative inference scores were highest in old-old adults. Participants over 65 years with the lowest scores on cognitive integrity variables also had significantly lower comprehension scores, but there was no effect of cognitive integrity on inference scores. This dissociation suggests that inferential processes may be maintained even when cognitive integrity and comprehension declines are present. Relevance to cognitive aging theories addressing text processing and self-regulatory processes is addressed. PMID- 19286644 TI - Self-rated health and depressive symptoms in patients with end-stage renal disease and their spouses: a longitudinal dyadic analysis of late-life marriages. AB - Limited research has examined the ways in which changes in self-rated health experienced by aging spouses affect depressive symptoms of both members of the dyad. Longitudinal data from 315 older couples in which one partner had end-stage renal disease were analyzed using multilevel modeling. Results indicate that for both patients and spouses, own mean self-rated health was associated with own depressive symptoms, and change in self-rated health had a significant negative association with change in own depressive symptoms. Both mean self-rated health of the patient and change in patient's self-rated health had negative relationships with spouse depressive symptoms, with changes in patient's self rated health having a stronger impact on spouse depressive symptoms than changes in spouse's own self-rated health. Results suggest the importance of understanding physical and mental health in the context of the marital dyad. PMID- 19286645 TI - Interracial roommate relationships: negotiating daily interactions. AB - Jobs, social group memberships, or living arrangements lead many people to interact every day with another person from a different racial background. Given that research has shown that interracial interactions are often stressful, it is important to know how these daily interactions unfold across time and what factors contribute to the success or failure of these interactions. Both members of same-race and mixed-race college roommate pairs completed daily questionnaires measuring their emotional experiences and their perceptions of their roommate. Results revealed that roommates in mixed-race dyads experienced less positive emotions and intimacy toward their roommates than did roommates in same-race dyads and that the experience of positive emotions declined over time for ethnic minority students with White roommates. Mediation analyses showed that the negative effects of roommate race were mediated by the level of intimacy-building behaviors performed by the roommate. Implications for future research and university policies are discussed. PMID- 19286646 TI - Serum amyloid A3 does not contribute to circulating SAA levels. AB - Adipose tissue secretes proteins like serum amyloid A (SAA), which plays important roles in local and systemic inflammation. Circulating SAA levels increase in obese humans, but the roles of adipose-derived SAA and hyperlipidemia in this process are unclear. We took advantage of the difference in the inducible isoforms of SAA secreted by adipose tissue (SAA3) and liver (SAA1 and 2) of mice to evaluate whether adipose tissue contributes to the circulating pool of SAA in obesity and hyperlipidemia. Genetically obese (ob/ob) mice, but not hyperlipidemic mice deficient in apolipoprotein E (Apoe(-/-)), had significantly higher circulating levels of SAA than their littermate controls. SAA1/2 mRNA expression in the liver and SAA3 mRNA expression in intra-abdominal fat were significantly higher in obese than thin mice, but they were not affected by hyperlipidemia in Apoe(-/-) mice. However, only SAA1/2 and the constitutive form of SAA (SAA4) could be detected in the circulation by mass spectrometric analysis of HDL, the major carrier of circulating SAA. In contrast, SAA3 could be detected in medium from cultured adipocytes. Our findings indicate that the expression of SAA3 in adipose tissue is upregulated by obesity, but it does not contribute to the circulating pool of SAA in mice. PMID- 19286647 TI - ABCA1 plays no role in the centripetal movement of cholesterol from peripheral tissues to the liver and intestine in the mouse. AB - This study uses the mouse to explore the role of ABCA1 in the movement of this cholesterol from the peripheral organs to the endocrine glands for hormone synthesis and liver for excretion. The sterol pool in all peripheral organs was constant and equaled 2,218 and 2,269 mg/kg, respectively, in abca1(+/+) and abca1(-/-) mice. Flux of cholesterol from these tissues equaled the rate of synthesis plus the rate of LDL-cholesterol uptake and was 49.9 mg/day/kg in control animals and 62.0 mg/day/kg in abca1(-/-) mice. In the abca1(+/+) animals, this amount of cholesterol moved from HDL into the liver for excretion. In the abca1(-/-) mice, the cholesterol from the periphery also reached the liver but did not use HDL. Fecal excretion of cholesterol was just as high in abac1(-/-) mice (198 mg/day/kg) as in the abac1(+/+) animals (163 mg/day/kg), although the abac1(-/-) mice excreted relatively more neutral than acidic sterols. This study established that ABCA1 plays essentially no role in the turnover of cholesterol in peripheral organs or in the centripetal movement of this sterol to the endocrine glands, liver, and intestinal tract for excretion. PMID- 19286648 TI - Adult-onset degeneration of adipose tissue in mice deficient for the Sox8 transcription factor. AB - Although the transcription factor Sox8 is broadly expressed during embryogenesis in developing ectodermal and mesodermal tissues, mice develop surprisingly normally in the absence of Sox8. Phenotypes in adult Sox8-deficient mice include mild osteopenia, late-onset male infertility, and reduced weight. We show here that progressive degeneration of adipose tissue in adult Sox8-deficient mice significantly contributes to weight reduction. Although serum levels of leptin, IGF-1, and noradrenaline were altered in Sox8-deficient mice, these changes could not explain the observed phenotype. Other serum parameters, including indicators of glucose metabolism, were largely normal. However, expression of the preadipocyte marker Pref-1 was elevated in adipose tissues of Sox8-deficient mice. This increase correlated with an impaired differentiation of Sox8-deficient fibroblasts to adipocytes in culture, a defect that could be rescued by reintroducing Sox8 into the cells. Furthermore, Sox8 levels were higher in mesodermal precursors than in mature adipocytes. We postulate a precursor intrinsic role of Sox8 during replenishment of the adipocyte pool in adult mice and assume that disturbance of this function significantly contributes to adipose tissue degeneration in Sox8-deficient mice. PMID- 19286649 TI - A novel direct homogeneous assay for ATP citrate lyase. AB - ATP citrate lyase (ACL) is a cytosolic enzyme that catalyzes the synthesis of acetyl-CoA and oxaloacetate using citrate, CoA, and ATP as substrates and Mg(2+) as a necessary cofactor. The ACL-dependent synthesis of acetyl-CoA is thought to be an essential step for the de novo synthesis of fatty acids and cholesterol. For this reason, inhibition of ACL has been pursued as a strategy to treat dyslipidemia and obesity. Traditionally, ACL enzyme activity is measured indirectly by coupling to enzymes such as malate dehydrogenase or chloramphenicol acetyl transferase. In this report, however, we describe a novel procedure to directly measure ACL enzyme activity. We first identified a convenient method to specifically detect [(14)C]acetyl-CoA without detecting [(14)C]citrate by MicroScint-O. Using this detection system, we devised a simple, direct, and homogeneous ACL assay in 384-well plate format that is suitable for high throughput screening. The current assay consists of 1) incubation of ACL enzyme with [(14)C]citrate and other substrates/cofactors CoA, ATP, and Mg(2+), 2) EDTA quench, 3) addition of MicroScint-O, the agent that specifically detects product [(14)C]acetyl-CoA, and 4) detection of signal by TopCount. This unique ACL assay may provide more efficient identification of new ACL inhibitors and allow detailed mechanistic characterization of ACL/inhibitor interactions. PMID- 19286650 TI - Arsenate reductase, mycothiol, and mycoredoxin concert thiol/disulfide exchange. AB - We identified the first enzymes that use mycothiol and mycoredoxin in a thiol/disulfide redox cascade. The enzymes are two arsenate reductases from Corynebacterium glutamicum (Cg_ArsC1 and Cg_ArsC2), which play a key role in the defense against arsenate. In vivo knockouts showed that the genes for Cg_ArsC1 and Cg_ArsC2 and those of the enzymes of the mycothiol biosynthesis pathway confer arsenate resistance. With steady-state kinetics, arsenite analysis, and theoretical reactivity analysis, we unraveled the catalytic mechanism for the reduction of arsenate to arsenite in C. glutamicum. The active site thiolate in Cg_ArsCs facilitates adduct formation between arsenate and mycothiol. Mycoredoxin, a redox enzyme for which the function was never shown before, reduces the thiol-arseno bond and forms arsenite and a mycothiol-mycoredoxin mixed disulfide. A second molecule of mycothiol recycles mycoredoxin and forms mycothione that, in its turn, is reduced by the NADPH-dependent mycothione reductase. Cg_ArsCs show a low specificity constant of approximately 5 m(-1) s( 1), typically for a thiol/disulfide cascade with nucleophiles on three different molecules. With the in vitro reconstitution of this novel electron transfer pathway, we have paved the way for the study of redox mechanisms in actinobacteria. PMID- 19286651 TI - Structural and functional dissection of the heterocyclic peptide cytotoxin streptolysin S. AB - The human pathogen Streptococcus pyogenes secretes a highly cytolytic toxin known as streptolysin S (SLS). SLS is a key virulence determinant and responsible for the beta-hemolytic phenotype of these bacteria. Despite over a century of research, the chemical structure of SLS remains unknown. Recent experiments have revealed that SLS is generated from an inactive precursor peptide that undergoes extensive post-translational modification to an active form. In this work, we address outstanding questions regarding the SLS biosynthetic process, elucidating the features of substrate recognition and sites of posttranslational modification to the SLS precursor peptide. Further, we exploit these findings to guide the design of artificial cytolytic toxins that are recognized by the SLS biosynthetic enzymes and others that are intrinsically cytolytic. This new structural information has ramifications for future antimicrobial therapies. PMID- 19286652 TI - Novel protective mechanism against irreversible hyperoxidation of peroxiredoxin: Nalpha-terminal acetylation of human peroxiredoxin II. AB - Peroxiredoxins (Prxs) are a group of peroxidases containing a cysteine thiol at their catalytic site. During peroxidase catalysis, the catalytic cysteine, referred to as the peroxidatic cysteine (C(P)), cycles between thiol (C(P)-SH) and disulfide (-S-S-) states via a sulfenic (C(P)-SOH) intermediate. Hyperoxidation of the C(P) thiol to its sulfinic (C(P)-SO(2)H) derivative has been shown to be reversible, but its sulfonic (C(P)-SO(3)H) derivative is irreversible. Our comparative study of hyperoxidation and regeneration of Prx I and Prx II in HeLa cells revealed that Prx II is more susceptible than Prx I to hyperoxidation and that the majority of the hyperoxidized Prx II formation is reversible. However, the hyperoxidized Prx I showed much less reversibility because of the formation of its irreversible sulfonic derivative, as verified with C(P)-SO(3)H-specific antiserum. In an attempt to identify the multiple hyperoxidized spots of the Prx I on two-dimensional PAGE analysis, an N acetylated Prx I was identified as part of the total Prx I using anti-acetylated Lys antibody. Using peptidyl-Asp metalloendopeptidase (EC 3.4.24.33) peptide fingerprints, we found that N(alpha)-terminal acetylation (N(alpha)-Ac) occurred exclusively on Prx II after demethionylation. N(alpha)-Ac of Prx II blocks Prx II from irreversible hyperoxidation without altering its affinity for hydrogen peroxide. A comparative study of non-N(alpha)-acetylated and N(alpha)-terminal acetylated Prx II revealed that N(alpha)-Ac of Prx II induces a significant shift in the circular dichroism spectrum and elevation of T(m) from 59.6 to 70.9 degrees C. These findings suggest that the structural maintenance of Prx II by N(alpha)-Ac may be responsible for preventing its hyperoxidation to form C(P) SO(3)H. PMID- 19286653 TI - Epigenetic control of the invasion-promoting MT1-MMP/MMP-2/TIMP-2 axis in cancer cells. AB - Membrane type-1 matrix metalloproteinase (MT1-MMP) is an activator of soluble MMP 2. The activity of both MMPs is regulated by their physiological inhibitor TIMP 2. An MT1-MMP/MMP-2/TIMP-2 axis plays a key role in the invasive behavior of many cell types. Despite its importance, epigenetic control of this pro-invasive axis is insufficiently studied, and, as a result, its modification in a rational and clinically beneficial manner is exceedingly difficult. Therefore, we performed an epigenetic analysis of the MT1-MMP, MMP-2, and TIMP-2 gene promoters in highly migratory glioblastoma cells and in low migratory breast carcinoma MCF-7 cells. We determined, for the first time, that the epigenetic control leading to the transcriptional silencing of both MMPs includes hypermethylation of the corresponding CpG regions and histone H3 lysine-27 trimethylation (H3K27me3). In turn, undermethylation of the CpG islands and low levels of histone H3 lysine-27 trimethylation are features of transcriptionally active MT1-MMP and MMP-2 genes in invasive cancer cells. Additional histone modifications we have analyzed, including H3ac and H3K4me2, are present in both transcriptionally active and inactive promoters of both MMPs. Histone H3 lysine-4 trimethylation is likely to play no significant role in regulating MT1-MMP and MMP-2. The pattern of epigenetic regulation of TIMP-2 was clearly distinct from that of MMPs and included the coordinated methylation and demethylation of the two CpG regions in the promoter. Our results suggest that the epigenetic control plays an important role in both the balanced regulation of the MT1-MMP/MMP-2/TIMP-2 axis and the invasive behavior in cancer cells. PMID- 19286654 TI - Ligand-specific conformational changes in the alpha1 glycine receptor ligand binding domain. AB - Understanding the activation mechanism of Cys loop ion channel receptors is key to understanding their physiological and pharmacological properties under normal and pathological conditions. The ligand-binding domains of these receptors comprise inner and outer beta-sheets and structural studies indicate that channel opening is accompanied by conformational rearrangements in both beta-sheets. In an attempt to resolve ligand-dependent movements in the ligand-binding domain, we employed voltage-clamp fluorometry on alpha1 glycine receptors to compare changes mediated by the agonist, glycine, and by the antagonist, strychnine. Voltage clamp fluorometry involves labeling introduced cysteines with environmentally sensitive fluorophores and inferring structural rearrangements from ligand induced fluorescence changes. In the inner beta-sheet, we labeled residues in loop 2 and in binding domain loops D and E. At each position, strychnine and glycine induced distinct maximal fluorescence responses. The pre-M1 domain responded similarly; at each of four labeled positions glycine produced a strong fluorescence signal, whereas strychnine did not. This suggests that glycine induces conformational changes in the inner beta-sheet and pre-M1 domain that may be important for activation, desensitization, or both. In contrast, most labeled residues in loops C and F yielded fluorescence changes identical in magnitude for glycine and strychnine. A notable exception was H201C in loop C. This labeled residue responded differently to glycine and strychnine, thus underlining the importance of loop C in ligand discrimination. These results provide an important step toward mapping the domains crucial for ligand discrimination in the ligand binding domain of glycine receptors and possibly other Cys loop receptors. PMID- 19286655 TI - Mismatch repair protein deficiency compromises cisplatin-induced apoptotic signaling. AB - Mismatch repair (MMR) proteins participate in cytotoxicity induced by certain DNA damage-inducing agents, including cisplatin (cis-diamminedichloroplatinum(II), CDDP), a cancer chemotherapeutic drug utilized clinically to treat a variety of malignancies. MMR proteins have been demonstrated to bind to CDDP-DNA adducts and initiate MMR protein-dependent cell death in cells treated with CDDP; however, the molecular events underlying this death remain unclear. As MMR proteins have been suggested to be important in clinical responses to CDDP, a clear understanding of MMR protein-dependent, CDDP-induced cell death is critical. In this report, we demonstrate MMR protein-dependent relocalization of cytochrome c to the cytoplasm and cleavage of caspase-9, caspase-3, and poly(ADP-ribose) polymerase upon treatment of cells with CDDP. Chemical inhibition of caspases specifically attenuates CDDP/MMR protein-dependent cytotoxicity, suggesting that a caspase-dependent signaling mechanism is required for the execution of this cell death. p53 protein levels were up-regulated independently of MMR protein status, suggesting that p53 is not a mediator of MMR-dependent, CDDP-induced death. This work is the first indication of a required signaling mechanism in CDDP-induced, MMR protein-dependent cytotoxicity, which can be uncoupled from other CDDP response pathways, and defines a critical contribution of MMR proteins to the control of cell death. PMID- 19286656 TI - Role of bound Zn(II) in the CadC Cd(II)/Pb(II)/Zn(II)-responsive repressor. AB - The Staphylococcus aureus plasmid pI258 cadCA operon encodes a P-type ATPase, CadA, that confers resistance to Cd(II)/Pb(II)/Zn(II). Expression is regulated by CadC, a homodimeric repressor that dissociates from the cad operator/promoter upon binding of Cd(II), Pb(II), or Zn(II). CadC is a member of the ArsR/SmtB family of metalloregulatory proteins. The crystal structure of CadC shows two types of metal binding sites, termed Site 1 and Site 2, and the homodimer has two of each. Site 1 is the physiological inducer binding site. The two Site 2 metal binding sites are formed at the dimerization interface. Site 2 is not regulatory in CadC but is regulatory in the homologue SmtB. Here the role of each site was investigated by mutagenesis. Both sites bind either Cd(II) or Zn(II). However, Site 1 has higher affinity for Cd(II) over Zn(II), and Site 2 prefers Zn(II) over Cd(II). Site 2 is not required for either derepression or dimerization. The crystal structure of the wild type with bound Zn(II) and of a mutant lacking Site 2 was compared with the SmtB structure with and without bound Zn(II). We propose that an arginine residue allows for Zn(II) regulation in SmtB and, conversely, a glycine results in a lack of regulation by Zn(II) in CadC. We propose that a glycine residue was ancestral whether the repressor binds Zn(II) at a Site 2 like CadC or has no Site 2 like the paralogous ArsR and implies that acquisition of regulatory ability in SmtB was a more recent evolutionary event. PMID- 19286657 TI - Mechanism of activation and functional role of protein kinase Ceta in human platelets. AB - The novel class of protein kinase C (nPKC) isoform eta is expressed in platelets, but not much is known about its activation and function. In this study, we investigated the mechanism of activation and functional implications of nPKCeta using pharmacological and gene knock-out approaches. nPKCeta was phosphorylated (at Thr-512) in a time- and concentration-dependent manner by 2MeSADP. Pretreatment of platelets with MRS-2179, a P2Y1 receptor antagonist, or YM 254890, a G(q) blocker, abolished 2MeSADP-induced phosphorylation of nPKCeta. Similarly, ADP failed to activate nPKCeta in platelets isolated from P2Y1 and G(q) knock-out mice. However, pretreatment of platelets with P2Y12 receptor antagonist, AR-C69331MX did not interfere with ADP-induced nPKCeta phosphorylation. In addition, when platelets were activated with 2MeSADP under stirring conditions, although nPKCeta was phosphorylated within 30 s by ADP receptors, it was also dephosphorylated by activated integrin alpha(IIb)beta3 mediated outside-in signaling. Moreover, in the presence of SC-57101, a alpha(IIb)beta3 receptor antagonist, nPKCeta dephosphorylation was inhibited. Furthermore, in murine platelets lacking PP1cgamma, a catalytic subunit of serine/threonine phosphatase, alpha(IIb)beta3 failed to dephosphorylate nPKCeta. Thus, we conclude that ADP activates nPKCeta via P2Y1 receptor and is subsequently dephosphorylated by PP1gamma phosphatase activated by alpha(IIb)beta3 integrin. In addition, pretreatment of platelets with eta-RACK antagonistic peptides, a specific inhibitor of nPKCeta, inhibited ADP-induced thromboxane generation. However, these peptides had no affect on ADP-induced aggregation when thromboxane generation was blocked. In summary, nPKCeta positively regulates agonist-induced thromboxane generation with no effects on platelet aggregation. PMID- 19286658 TI - Intracellular transport of human immunodeficiency virus type 1 genomic RNA and viral production are dependent on dynein motor function and late endosome positioning. AB - Our earlier work indicated that the human immunodeficiency virus type 1 (HIV-1) genomic RNA (vRNA) is trafficked to the microtubule-organizing center (MTOC) when heterogeneous nuclear ribonucleoprotein A2/B1 is depleted from cells. Also, Rab7 interacting lysosomal protein promoted dynein motor complex, late endosome and vRNA clustering at the MTOC suggesting that the dynein motor and late endosomes were involved in vRNA trafficking. To investigate the role of the dynein motor in vRNA trafficking, dynein motor function was disrupted by small interference RNA mediated depletion of the dynein heavy chain or by p50/dynamitin overexpression. These treatments led to a marked relocalization of vRNA and viral structural protein Gag to the cell periphery with late endosomes and a severalfold increase in HIV-1 production. In contrast, rerouting vRNA to the MTOC reduced virus production. vRNA localization depended on Gag membrane association as shown using both myristoylation and Gag nucleocapsid domain proviral mutants. Furthermore, the cytoplasmic localization of vRNA and Gag was not attributable to intracellular or internalized endocytosed virus particles. Our results demonstrate that dynein motor function is important for regulating Gag and vRNA egress on endosomal membranes in the cytoplasm to directly impact on viral production. PMID- 19286659 TI - GTP cyclohydrolase I expression, protein, and activity determine intracellular tetrahydrobiopterin levels, independent of GTP cyclohydrolase feedback regulatory protein expression. AB - GTP cyclohydrolase I (GTPCH) is a key enzyme in the synthesis of tetrahydrobiopterin (BH4), a required cofactor for nitricoxide synthases and aromatic amino acid hydroxylases. Alterations of GTPCH activity and BH4 availability play an important role in human disease. GTPCH expression is regulated by inflammatory stimuli, in association with reduced expression of GTP cyclohydrolase feedback regulatory protein (GFRP). However, the relative importance of GTPCH expression versus GTPCH activity and the role of GFRP in relation to BH4 bioavailability remain uncertain. We investigated these relationships in a cell line with tet-regulated GTPCH expression and in the hph-1 mouse model of GTPCH deficiency. Doxycycline exposure resulted in a dose dependent decrease in GTPCH protein and activity, with a strong correlation between GTPCH expression and BH4 levels (r(2) = 0.85, p < 0.0001). These changes in GTPCH and BH4 had no effect on GFRP expression or protein levels. GFRP overexpression and knockdown in tet-GCH cells did not alter GTPCH activity or BH4 levels, and GTPCH-specific knockdown in sEnd.1 endothelial cells had no effect on GFRP protein. In mouse liver we observed a graded reduction of GTPCH expression, protein, and activity, from wild type, heterozygote, to homozygote littermates, with a striking linear correlation between GTPCH expression and BH4 levels (r(2) = 0.82, p < 0.0001). Neither GFRP expression nor protein differed between wild type, heterozygote, nor homozygote mice, despite the substantial differences in BH4. We suggest that GTPCH expression is the primary regulator of BH4 levels, and changes in GTPCH or BH4 are not necessarily accompanied by changes in GFRP expression. PMID- 19286660 TI - Neutral cysteine protease bleomycin hydrolase is essential for the breakdown of deiminated filaggrin into amino acids. AB - Filaggrin is a component of the cornified cell envelope and the precursor of free amino acids acting as a natural moisturizing factor in the stratum corneum. Deimination is critical for the degradation of filaggrin into free amino acids. In this study, we tried to identify the enzyme(s) responsible for the cleavage of deiminated filaggrin in vitro. First, we investigated citrulline aminopeptidase activity in the extract of newborn rat epidermis by double layer fluorescent zymography and detected strong activity at neutral pH. Monitoring the citrulline releasing activity, we purified an enzyme of 280 kDa, comprised of six identical subunits of 48 kDa. The NH(2) terminus of representative tryptic peptides perfectly matched the sequence of rat bleomycin hydrolase (BH). The enzyme released various amino acids except Pro from beta-naphthylamide derivatives and hydrolyzed citrulline-beta-naphthylamide most effectively. Thus, to break down deiminated filaggrin, another protease would be required. Among proteases tested, calpain I degraded the deiminated filaggrin effectively into many peptides of different mass on the matrix-assisted laser desorption/ionization-time of flight mass spectrum. We confirmed that various amino acids including citrulline were released by BH from those peptides. On the other hand, caspase 14 degraded deiminated filaggrin into a few peptides of limited mass. Immunohistochemical analysis of normal human skin revealed co-localization of BH and filaggrin in the granular layer. Collectively, our results suggest that BH is essential for the synthesis of natural moisturizing factors and that calpain I would play a role as an upstream protease in the degradation of filaggrin. PMID- 19286661 TI - Plasmin-mediated proteolysis is required for hepatocyte growth factor activation during liver repair. AB - The physiological relevance of the activation of hepatocyte growth factor (Hgf) by the plasminogen (Plg) system of proteases and its contribution to tissue repair are largely undefined. Here, we investigated whether the defective liver repair in mice lacking Plg is due to impaired activation of Hgf. Loss of Plg in vivo suppressed Hgf activation and signaling through its Met tyrosine kinase receptor. Without Plg, hepatocytes were unresponsive to Hgf-induced proliferation and migration, with a more pronounced impairment in hepatocyte movement within the hepatic environment. Most notably, circumventing the defect in proteolytic activation of Hgf by the downstream expression of an activated Met receptor corrected the functional deficits and improved liver repair in Plg-deficient mice. These findings support a fibrinolysis-unrelated role for Plg in modulating cell proliferation and migration by activation of Hgf. PMID- 19286662 TI - Lipid G protein-coupled receptor ligand identification using beta-arrestin PathHunter assay. AB - A growing number of orphan G-protein-coupled receptors (GPCRs) have been reported to be activated by lipid ligands, such as lysophosphatidic acid, sphingosine 1 phosphate (S1P), and cannabinoids, for which there are already well established receptors. These new ligand claims are controversial due to either lack of independent confirmations or conflicting reports. We used the beta-arrestin PathHunter assay system, a newly developed, generic GPCR assay format that measures beta-arrestin binding to GPCRs, to evaluate lipid receptor and ligand pairing. This assay eliminates interference from endogenous receptors on the parental cells because it measures a signal that is specifically generated by the tagged receptor and is immediately downstream of receptor activation. We screened a large number of newly "deorphaned" receptors (GPR23, GPR92, GPR55, G2A, GPR18, GPR3, GPR6, GPR12, and GPR63) and control receptors against a collection of approximately 400 lipid molecules to try to identify the receptor ligand in an unbiased fashion. GPR92 was confirmed to be a lysophosphatidic acid receptor with weaker responses to farnesyl pyrophosphate and geranylgeranyl diphosphate. The putative cannabinoid receptor GPR55 responded strongly to AM251, rimonabant, and lysophosphatidylinositol but only very weakly to endocannabinoids. G2A receptor was confirmed to be an oxidized free fatty acid receptor. In addition, we discovered that 3,3'-diindolylmethane, a dietary molecule from cruciferous vegetables, which has known anti-cancer properties, to be a CB(2) receptor partial agonist, with binding affinity around 1 microm. The anti-inflammatory effect of 3,3'-diindolylmethane in RAW264.7 cells was shown to be partially mediated by CB(2). PMID- 19286663 TI - Direct magnetic resonance evidence for peroxymonocarbonate involvement in the cu,zn-superoxide dismutase peroxidase catalytic cycle. AB - Cu,Zn-superoxide dismutase (SOD1) is a copper- and zinc-dependent enzyme. The main function of SOD1 is believed to be the scavenging and detoxification of superoxide radicals. Nevertheless, the last 30 years have seen a rapid accumulation of evidence indicating that SOD1 may also act as a peroxidase, an alternative function that was implicated in the onset and progression of familial amyotrophic lateral sclerosis. Although SOD1 peroxidase activity and its dependence on carbon dioxide have been well described, the molecular basis of the SOD1 peroxidase cycle remains obscure, because none of the proposed catalytic intermediates have so far been identified. In view of recent observations, we hypothesized that the SOD1 peroxidase cycle relies on two steps: 1) reduction of SOD-Cu(II) by hydrogen peroxide followed by 2) oxidation of SOD-Cu(I) by peroxymonocarbonate, the product of the spontaneous reaction of bicarbonate with hydrogen peroxide, to produce SOD-Cu(II) and carbonate radical anion. This hypothesis has been investigated through electron paramagnetic resonance and nuclear magnetic resonance to provide direct evidence for a peroxycarbonate driven, SOD1-catalyzed carbonate radical production. The results gathered herein indicate that peroxymonocarbonate (HOOCO(2)(-)) is a key intermediate in the SOD1 peroxidase cycle and identify this species as the precursor of carbonate radical anions. PMID- 19286664 TI - Turning cancer stem cells inside out: an exploration of glioma stem cell signaling pathways. AB - Tumors are complex collections of heterogeneous cells with recruited vasculature, inflammatory cells, and stromal elements. Neoplastic cells frequently display a hierarchy in differentiation status. Recent studies suggest that brain tumors have a limited population of neoplastic cells called cancer stem cells with the capacity for sustained self-renewal and tumor propagation. Brain tumor stem cells contribute to therapeutic resistance and tumor angiogenesis. In this minireview, we summarize recent data regarding critical signaling pathways involved in brain tumor stem cell biology and discuss how targeting these molecules may contribute to the development of novel anti-glioma therapies. PMID- 19286665 TI - Identification of PpoA from Aspergillus nidulans as a fusion protein of a fatty acid heme dioxygenase/peroxidase and a cytochrome P450. AB - The homothallic ascomycete Aspergillus nidulans serves as model organism for filamentous fungi because of its ability to propagate with both asexual and sexual life cycles, and fatty acid-derived substances regulate the balance between both cycles. These so-called psi (precocious sexual inducer) factors are produced by psi factor-producing oxygenases (Ppo enzymes). Bioinformatic analysis predicted the presence of two different heme domains in Ppo proteins: in the N terminal region, a fatty acid heme dioxygenase/peroxidase domain is predicted, whereas in the C-terminal region, a P450 heme thiolate domain is predicted. To analyze the reaction catalyzed by Ppo enzymes, PpoA was expressed in Escherichia coli as an active enzyme. The protein was purified by 62-fold and identified as a homotetrameric ferric heme protein that metabolizes mono- as well as polyunsaturated C(16) and C(18) fatty acids at pH approximately 7.25. The presence of thiolate-ligated heme was confirmed on the basis of sequence alignments and the appearance of a characteristic 450 nm CO-binding spectrum. Studies on its reaction mechanism revealed that PpoA uses different heme domains to catalyze two separate reactions. Within the heme peroxidase domain, linoleic acid is oxidized to (8R)-hydroperoxyoctadecadienoic acid by abstracting a H-atom from C-8 of the fatty acid, yielding a carbon-centered radical that reacts with molecular dioxygen. In the second reaction step, 8-hydroperoxyoctadecadienoic acid is isomerized within the P450 heme thiolate domain to 5,8 dihydroxyoctadecadienoic acid. We identify PpoA as a bifunctional P450 fusion protein that uses a previously unknown reaction mechanism for forming psi factors. PMID- 19286666 TI - Chromosome end maintenance by telomerase. AB - Telomeres, protein-DNA complexes at the ends of eukaryotic linear chromosomes, are essential for genome stability. The accumulation of chromosomal abnormalities in the absence of proper telomere function is implicated in human aging and cancer. Repetitive telomeric sequences are maintained by telomerase, a ribonucleoprotein complex containing a reverse transcriptase subunit, a template RNA, and accessory components. Telomere elongation is regulated at multiple levels, including assembly of the telomerase holoenzyme, recruitment of telomerase to the chromosome terminus, and telomere accessibility. This minireview provides an overview of telomerase structure, function, and regulation and the role of telomerase in human disease. PMID- 19286667 TI - Tetrahydrobiopterin recycling, a key determinant of endothelial nitric-oxide synthase-dependent signaling pathways in cultured vascular endothelial cells. AB - Tetrahydrobiopterin (BH4) is a key redox-active cofactor in endothelial isoform of NO synthase (eNOS) catalysis and is an important determinant of NO-dependent signaling pathways. BH4 oxidation is observed in vascular cells in the setting of the oxidative stress associated with diabetes. However, the relative roles of de novo BH4 synthesis and BH4 redox recycling in the regulation of eNOS bioactivity remain incompletely defined. We used small interference RNA (siRNA)-mediated "knockdown" GTP cyclohydrolase-1 (GTPCH1), the rate-limiting enzyme in BH4 biosynthesis, and dihydrofolate reductase (DHFR), an enzyme-recycling oxidized BH4 (7,8-dihydrobiopterin (BH2)), and studied the effects on eNOS regulation and biopterin metabolism in cultured aortic endothelial cells. Knockdown of either DHFR or GTPCH1 attenuated vascular endothelial growth factor (VEGF)-induced eNOS activity and NO production; these effects were recovered by supplementation with BH4. In contrast, supplementation with BH2 abolished VEGF-induced NO production. DHFR but not GTPCH1 knockdown increased reactive oxygen species (ROS) production. The increase in ROS production seen with siRNA-mediated DHFR knockdown was abolished either by simultaneous siRNA-mediated knockdown of eNOS or by supplementing with BH4. In contrast, addition of BH2 increased ROS production; this effect of BH2 was blocked by BH4 supplementation. DHFR but not GTPCH1 knockdown inhibited VEGF-induced dephosphorylation of eNOS at the inhibitory site serine 116; these effects were recovered by supplementation with BH4. These studies demonstrate a striking contrast in the pattern of eNOS regulation seen by the selective modulation of BH4 salvage/reduction versus de novo BH4 synthetic pathways. Our findings suggest that the depletion of BH4 is not sufficient to perturb NO signaling, but rather that concentration of intracellular BH2, as well as the relative concentrations of BH4 and BH2, together play a determining role in the redox regulation of eNOS-modulated endothelial responses. PMID- 19286668 TI - Identification of interactive networks of gene expression associated with osteosarcoma oncogenesis by integrated molecular profiling. AB - Altered gene expression in tumors can be caused by copy number alterations to DNA or mutation affecting coding or regulatory regions of genes. However, epigenetic events may also influence gene expression. Malignant cells can show major disruptions in DNA methylation profiles, which are manifested as aberrant hypermethylation or as hypomethylation of gene promoters, as well as global genomic hypomethylation. In this study we performed integrative whole-genome analysis of DNA copy number, promoter methylation and gene expression using 10 osteosarcomas. We identified significant changes including: hypomethylation, gain, and overexpression of histone cluster 2 genes at chromosome 1q21.1-q21.3; loss of chromosome 8p21.2-p21.3 and underexpression of DOCK5 and TNFRSF10A/D genes; and amplification-related overexpression of RUNX2 at chromosome 6p12.3 p21.1. Amplification and overexpression of RUNX2 could disrupt G2/M cell cycle checkpoints, and downstream osteosarcoma-specific changes, such as failure of bone differentiation and genomic polyploidization. Failure of DOCK5-signaling, together with p53 and TNFRSF10A/D-related cell cycle and death pathways, may play a critical role in abrogating apoptosis. Our analyses show that the RUNX2 interactome may be constitutively activated in osteosarcoma, and that the downstream intracellular pathways are strongly associated with the regulation of osteoblast differentiation and control of cell cycle and apoptosis in osteosarcoma. PMID- 19286669 TI - Attenuated muscle regeneration is a key factor in dysferlin-deficient muscular dystrophy. AB - Skeletal muscle requires an efficient and active membrane repair system to overcome the rigours of frequent contraction. Dysferlin is a component of that system and absence of dysferlin causes muscular dystrophy (dysferlinopathy) characterized by adult onset muscle weakness, high serum creatine kinase levels and a prominent inflammatory infiltrate. We have observed that dysferlinopathy patient biopsies show an excess of immature fibres and therefore investigated the role of dysferlin in muscle regeneration. Using notexin-induced muscle damage, we have shown that regeneration is attenuated in a mouse model of dysferlinopathy, with delayed removal of necrotic fibres, an extended inflammatory phase and delayed functional recovery. Satellite cell activation and myoblast fusion appear normal, but there is a reduction in early neutrophil recruitment in regenerating and also needle wounded muscle in dysferlin-deficient mice. Primary mouse dysferlinopathy myoblast cultures show reduced cytokine release upon stimulation, indicating that the secretion of chemotactic molecules is impaired. We suggest an extension to the muscle membrane repair model, where in addition to fusing patch repair vesicles with the sarcolemma dysferlin is also involved in the release of chemotactic agents. Reduced neutrophil recruitment results in incomplete cycles of regeneration in dysferlinopathy which combines with the membrane repair deficit to ultimately trigger dystrophic pathology. This study reveals a novel pathomechanism affecting muscle regeneration and maintenance in dysferlinopathy and highlights enhancement of the neutrophil response as a potential therapeutic avenue in these disorders. PMID- 19286670 TI - The STAT4 gene influences the genetic predisposition to systemic sclerosis phenotype. AB - The aim of this study was to investigate the possible role of STAT4 gene in the genetic predisposition to systemic sclerosis (SSc) susceptibility or clinical phenotype. A total of 1317 SSc patients [896 with limited cutaneous SSc (lcSSc) and 421 with diffuse cutaneous SSc (dcSSc)] and 3113 healthy controls, from an initial case-control set of Spanish Caucasian ancestry and five independent cohorts of European ancestry (The Netherlands, Germany, Sweden, Italy and USA), were included in the study. The rs7574865 polymorphism was selected as STAT4 genetic marker. We observed that the rs7574865 T allele was significantly associated with susceptibility to lcSSc in the Spanish population [P = 1.9 x 10( 5) odds ratio (OR) 1.61 95% confidence intervals (CI) 1.29-1.99], but not with dcSSc (P = 0.41 OR 0.84 95% CI 0.59-1.21). Additionally, a dosage effect was observed showing individuals with rs7574865 TT genotype higher risk for lcSSc (OR 3.34, P = 1.02 x 10(-7) 95% CI 2.11-5.31). The association of the rs7574865 T allele with lcSSc was confirmed in all the replication cohorts with different effect sizes (OR ranging between 1.15 and 1.86), as well as the lack of association of STAT4 with dcSSc. A meta-analysis to test the overall effect of the rs7574865 polymorphism showed a strong risk effect of the T allele for lcSSc susceptibility (pooled OR 1.54 95% CI 1.36-1.74; P < 0.0001). Our data show a strong and reproducible association of the STAT4 gene with the genetic predisposition to lcSSc suggesting that this gene seems to be one of the genetic markers influencing SSc phenotype. PMID- 19286671 TI - Pathway and network-based analysis of genome-wide association studies in multiple sclerosis. AB - Genome-wide association studies (GWAS) testing several hundred thousand SNPs have been performed in multiple sclerosis (MS) and other complex diseases. Typically, the number of markers in which the evidence for association exceeds the genome wide significance threshold is very small, and markers that do not exceed this threshold are generally neglected. Classical statistical analysis of these datasets in MS revealed genes with known immunological functions. However, many of the markers showing modest association may represent false negatives. We hypothesize that certain combinations of genes flagged by these markers can be identified if they belong to a common biological pathway. Here we conduct a pathway-oriented analysis of two GWAS in MS that takes into account all SNPs with nominal evidence of association (P < 0.05). Gene-wise P-values were superimposed on a human protein interaction network and searches were conducted to identify sub-networks containing a higher proportion of genes associated with MS than expected by chance. These sub-networks, and others generated at random as a control, were categorized for membership of biological pathways. GWAS from eight other diseases were analyzed to assess the specificity of the pathways identified. In the MS datasets, we identified sub-networks of genes from several immunological pathways including cell adhesion, communication and signaling. Remarkably, neural pathways, namely axon-guidance and synaptic potentiation, were also over-represented in MS. In addition to the immunological pathways previously identified, we report here for the first time the potential involvement of neural pathways in MS susceptibility. PMID- 19286672 TI - A novel interaction between fibroblast growth factor receptor 3 and the p85 subunit of phosphoinositide 3-kinase: activation-dependent regulation of ERK by p85 in multiple myeloma cells. AB - Ectopic activation of fibroblast growth factor receptor 3 (FGFR3) is associated with several cancers, including multiple myeloma (MM). FGFR3 inhibition in these cells inhibits proliferation and induces apoptosis, validating FGFR3 signaling as a therapeutic target in t(4;14) MM cases. We have identified the PI3K regulatory subunit, p85alpha, as a novel interactor of FGFR3 by yeast two-hybrid, and confirmed an interaction with both p85alpha and p85beta in mammalian cells. The interaction of FGFR3 with p85 is dependent upon receptor activation. In contrast to the Gab1-mediated association of FGFRs with p85, the FGFR3-p85 interaction we observed requires FGFR3 Y760, previously identified as a PLCgamma binding site. The interaction of p85 with FGFR3 does not require PLCgamma, suggesting the p85 interaction is direct and independent of PLCgamma binding. FGFR3 and p85 proteins also interact in MM cell lines which consistently express p85alpha and p85beta, but not p50 or p55 subunits. siRNA knockdown of p85beta in MM cells caused an increased ERK response to FGF2. These data suggest that an endogenous negative regulatory role for the p85-FGFR3 interaction on the Ras/ERK/MAPK pathway may exist in response to FGFR3 activity and identifies a novel therapeutic target for MM. PMID- 19286673 TI - ITGAM is associated with disease susceptibility and renal nephritis of systemic lupus erythematosus in Hong Kong Chinese and Thai. AB - ITGAM was recently found to be associated with systemic lupus erythematosus (SLE) in populations of not only European ancestry, but also in Hispanic- and African Americans, Mexicans and Colombians. The risk alleles in the gene, however, were found to be monomorphic in two Asian populations examined: Japanese and Korean. In this study, using a collection of 910 SLE patients and 2360 controls from Chinese living in Hong Kong, analyzed by both genome-wide association and direct sequencing, we confirmed the association of the same risk alleles in ITGAM with the disease. These findings were further replicated in the Thai population with 278 patients and 383 ethnicity- and geography-matched controls. Subphenotype stratification analyses showed significantly more involvement of the gene in patients with renal nephritis and neurological disorders. Although our results support a pivotal role by rs1143679 (R77H) in disease association, our data also suggests an additional contribution from rs1143683, another non-synonymous polymorphism in this gene (A858V). Therefore, despite the low-allele frequencies of the risk alleles of the gene in our two Asian populations, ITGAM was confirmed to be a risk factor related to disease susceptibility and probably severe manifestations of SLE. PMID- 19286674 TI - Tubby-like protein 3 (TULP3) regulates patterning in the mouse embryo through inhibition of Hedgehog signaling. AB - Tubby-like protein 3 (TULP3) is required for proper embryonic development in mice. Disruption of mouse Tulp3 results in morphological defects in the embryonic craniofacial regions, the spinal neural tube and the limbs. Here, we show that TULP3 functions as a novel negative regulator of Sonic hedgehog (Shh) signaling in the mouse. In Tulp3 mutants, ventral cell types in the lumbar neural tube, which acquire their identities in response to Shh signaling, are ectopically specified at the expense of dorsal cell types. Genetic epistasis experiments show that this ventralized phenotype occurs independently of Shh and the transmembrane protein Smoothened, but it is dependent on the transcription factor Gli2. The ventralized phenotype is also dependent on the kinesin II subunit Kif3A, which is required for intraflagellar transport and ciliogenesis. In addition, TULP3 is required for proper Shh-dependent limb patterning and for maintaining the correct balance between differentiation and proliferation in the neural tube. Finally, the localization of TULP3 to the tips of primary cilia raises the possibility that it regulates the Hedgehog pathway within this structure. PMID- 19286675 TI - Predictors of left ventricular remodelling and failure in right ventricular pacing in the young. AB - AIMS: To identify risk factors for left ventricular (LV) dysfunction in right ventricular (RV) pacing in the young. methods and results: Left ventricular function was evaluated in 82 paediatric patients with either non-surgical (n = 41) or surgical (n= 41) complete atrioventricular block who have been 100% RV paced for a mean period of 7.4 years. Left ventricular shortening fraction (SF) decreased from a median (range) of 39 (24-62)% prior to implantation to 32 (8 49)% at last follow-up (P < 0.05). Prevalence of a combination of LV dilatation (LV end-diastolic diameter >+2z-values) and dysfunction (SF < 0.26) was found to increase from 1.3% prior to pacemaker implantation to 13.4% (11/82 patients) at last follow-up (P = 0.01). Ten of these 11 patients had progressive LV remodelling and 8 of 11 were symptomatic. The only significant risk factor for the development of LV dilatation and dysfunction was the presence of epicardial RV free wall pacing (OR = 14.3, P < 0.001). Other pre-implantation demographic, diagnostic, and haemodynamic factors including block aetiology, pacing variables, and pacing duration did not show independent significance. CONCLUSION: Right ventricular pacing leads to pathologic LV remodelling in a significant proportion of paediatric patients. The major independent risk factor is the presence of epicardial RV free wall pacing, which should be avoided whenever possible. PMID- 19286676 TI - Use of emergency departments by older people from residential care: a population based study. AB - OBJECTIVE: to investigate the differences between emergency department (ED) presentations of older people who do and do not live in residential care facilities (RCFs). DESIGN: a population-based retrospective cohort study. ED records linked to ambulance and hospital morbidity records. SETTING: public EDs in Perth, Western Australia. SUBJECTS: all patients 65 years and older who presented to EDs between 1 January 2003 and 31 December 2006 (n = 97,161). MEASUREMENTS: patient demographic and clinical characteristics. Logistic regression was used to model the effect of living in RCFs on the likelihood of a particular ED diagnosis, hospital admission and in-hospital death. RESULTS: the age-sex standardised rate of ED presentation for those living in RCFs was 1.69 times that of community dwellers. Compared to community dwellers, people from RCFs were older (mean age 84.7 years vs. 76.0 years, P < 0.001) and a lower proportion were male (28.7% vs. 46.7%, P < 0.001). Adjusting for age and sex revealed that people from RCFs were more likely to be diagnosed with pneumonia/influenza [odds ratio (OR) 1.94, 95% confidence interval (CI) = 1.72 2.19], urinary tract infections (OR 1.72, 95% CI 1.49-1.98) or hip fractures (OR 1.16, 95% CI 1.03-1.32); less likely to be diagnosed with circulatory system diseases (OR 0.69, 95% CI 0.64-0.75) or neoplasms (OR 0.47, 95% CI 0.31-0.72); more likely to be admitted (OR 1.13, 95% CI 1.06-1.20) and to die in hospital (OR 1.57, 95% CI 1.40-1.75). CONCLUSION: there are different patterns of ED presentations and hospital admissions of older people who do and do not live in RCFs. The appropriateness of these differences is uncertain. PMID- 19286677 TI - Is vascular stiffness associated with the diameter of the abdominal aorta? PMID- 19286678 TI - Pseudoperipheral palsy due to cortical infarction. AB - We present two cases of isolated hand weakness that were initially thought to be due to lower motor neurone lesions until subsequent MRI confirmed strokes in both. Small strokes are important to identify in order to optimise secondary prevention. PMID- 19286679 TI - Severity of alcohol dependence in decompensated alcoholic liver disease: comparison with heavy drinkers without liver disease and relationship to family drinking history. AB - AIM: The aim of this study was to compare alcohol dependence severity in patients with severe alcoholic liver disease (ALD) with that in heavy drinkers without liver disease. METHODS: Short alcohol dependence data and lifetime alcohol questionnaires applied to unselected heavy alcohol drinkers (>60 units/week (M) or 40 units/week (F) for >5 years) with either (a) decompensated ALD (patients n = 136) or (b) no evidence of serious liver disease by clinical, biochemical and ultrasound evaluation ('controls' n = 148). RESULTS: The SADD alcohol dependence severity score (range 0-42) in patients with ALD was >28 (severe dependence) in 36 cases (26%); slightly higher than that in heavy-drinking controls taken as a whole; similar to that in controls who were seeking healthcare but higher than that in controls who were not; and lower than that in controls who attended specialist alcohol services. In ALD patients and controls, the SADD score was higher in those with three or more heavy-drinking first-degree relatives than in those with none. In multiple regression analysis, the SADD score showed independent associations with young age, clinically manifest alcohol dependence, seeking healthcare and the presence of multiple heavy drinking relatives, but not with ALD. CONCLUSIONS: Alcohol dependence severity in patients with ALD varies and tends to be lower than that in heavy drinkers seeking treatment at alcohol treatment centres but is not as low as implied in some previous studies. Alcohol dependence severity is associated with young age and family drinking history but is not specifically associated with the development of liver disease. PMID- 19286680 TI - Successful transapical aortic valve implantation four weeks before 97th birthday. AB - Aortic valve replacement (AVR) is currently the treatment of choice in patients with symptomatic aortic stenosis, but transcatheter off-pump aortic valve implantation is a novel and emerging technique, which might be performed more frequently in the near future and may be a less invasive alternative to conventional AVR in selected high-risk patients. Up to date, the preoperative patient selection is debated and potential candidates for these new techniques are widely discussed. We report a successful transapical aortic valve implantation performed in a 96-year-old woman demonstrating the potential of the novel technique as an alternative treatment option in old and multimorbid patients at high risk for conventional AVR. PMID- 19286681 TI - Intraoperative heart rate variability of a cardiac surgeon himself in coronary artery bypass grafting surgery. AB - The mental strain of a cardiac surgeon may differ when he performs coronary surgery from and when he only assists in performing coronary surgery. In 50 selected cases of on-pump heart arrested coronary artery bypass grafting (CABG), an attending-consultant surgeon performed 30 procedures of CABG (Group A) and an attending-consultant surgeon supervised the remaining 20 cases of CABG performed by two resident surgeons (Group B). Intraoperative Holter electrocardiograms of the attending-consultant surgeon were recorded and analyzed for heart rate variability (HRV). In Group A, the ratio of low frequency to high frequency was at a peak in the beginning of the operation and gradually decreased toward the end of the operation. In Group B, the ratio of low frequency to high frequency was at a peak in the phase of aortic cross-clamp, coronary anastomosis, and unclamping. When an attending-consultant surgeon performed the operation himself, the most anxious part of the operation was at the beginning and thereafter the level of anxiety gradually declined. In contrast, when he assisted a resident, the highest level of anxiety was when the aortic cross-clamp was in place and out of place and during the coronary anastomosis. PMID- 19286682 TI - Image-guided surgical repair of ventricular septal rupture using self-expanding device. AB - The purpose of this study was to determine the possibility of a new surgical technique for the treatment of ventricular septal rupture after acute myocardial infarction. The operations were conducted under the guidance of real-time three dimensional echocardiography (RT3DE) (iE33, Philips Medical Systems, Andover, MA). Six pigs were anesthetized, and after median sternotomy, the echo probe was applied directly to the surface of the heart. A ventricular septal defect (VSD) was created in all the six porcine hearts. The VSDs were closed with an Amplatzer septal occluder (AGA Medical Corp, Golden Valley, MN) through the right ventricular free wall under RT3DE monitoring. The procedure was successful in all the six pigs. The VSDs were precisely closed with the septal occluder under RT3DE guidance. Both left ventriculography and color-Doppler echocardiography showed no residual shunt in any of the six pigs. The use of the Amplatzer septal occluder under the guidance of real-time three-dimensional echocardiography made it possible to close the ventricular septal defect safely and successfully without the need to resort to cardiopulmonary bypass. Our results strongly suggest that the application of this new technique is feasible for the treatment of ventricular septal rupture after acute myocardial infarction. PMID- 19286683 TI - A case-crossover study on transient risk factors of work-related eye injuries. AB - OBJECTIVES: To investigate modifiable risk and preventive factors of work-related eye injuries. METHODS: A case-crossover study conducted to explore the associations between transient risk factors and work-related eye injuries. Patients seen at seven medical centres in Taiwan with work-related eye injuries over a 4-year period were enrolled in the study. Clinical information was collected from medical charts and detailed information on exposure to eight potentially modifiable factors during the 60 minutes prior to the occurrence of each injury, as well as during the same time interval on the last work day prior to the injury, were obtained using questionnaire surveys. Matched-pair interval analysis was adopted to assess the odds ratios (ORs) for work-related eye injuries given exposure to the eight modifiable factors. RESULTS: A total of 283 subjects were interviewed. Most of these injured workers were young, male, and self-employed or small enterprise workers. The most common injury type was photokeratitis (33.2%), mainly caused by welding (30.4%). The OR for a work related eye injury was increased with the performance of an unfamiliar task (57.0), operation of a faulty tool or piece of equipment (48.5), distractions (24.0), being rushed (13.0), or fatigued (10.0), and a poor work environment (4.3). Wearing eye protection devices was found to have a significant protective effect on workers who might otherwise have been exposed to eye injuries (OR = 0.4; 95% CI 0.2 to 0.7). CONCLUSION: Potential modifiable risk and preventive factors for work-related eye injuries were identified using a case-crossover study. This information should be helpful in the development of preventive strategies. PMID- 19286684 TI - Occupational exposure to potential endocrine disruptors: further development of a job exposure matrix. AB - OBJECTIVES: The aim was to develop a new up-to-date and comprehensive job exposure matrix (JEM) for estimating exposure to potential endocrine disruptors in epidemiological research. METHODS: Chemicals with endocrine disrupting properties were identified from the literature and classified into 10 chemical groups: polycyclic aromatic hydrocarbons (PAHs), polychlorinated organic compounds, pesticides, phthalates, organic solvents, bisphenol A, alkylphenolic compounds, brominated flame retardants, metals and a miscellaneous group. Most chemical groups were divided into three to six subgroups. Focusing on the years 1996-2006, three experts scored the probability of exposure to each chemical group and subgroup for 353 job titles as "unlikely" (0), "possible" (1) or "probable" (2). Job titles with positive exposure probability scores were provided with exposure scenarios that described the reasoning behind the scores. RESULTS: Exposure to any chemical group was unlikely for 238 job titles (67%), whereas 102 (29%) job titles were classified as possibly (17%) or probably (12%) exposed to one or several endocrine disruptors. The remaining 13 job titles provided too little information to classify exposure. PAHs, pesticides, phthalates, organic solvents, alkylphenolic compounds and metals were often linked to a job title in the JEM. The remaining chemical groups were found to involve very few occupations. CONCLUSIONS: Despite some important limitations, this JEM could be a valuable tool for exposure assessment in studies on the health risks of endocrine disruptors, especially when task specific information is incorporated. The documented exposure scenarios are meant to facilitate further adjustments to the JEM to allow more widespread use. PMID- 19286685 TI - Paraneoplastic retinopathy with multiple detachments of the neurosensory retina and autoantibodies against interphotoreceptor retinoid binding protein (IRBP) in cutaneous melanoma. PMID- 19286686 TI - Impediments to eye transplantation: ocular viability following optic-nerve transection or enucleation. AB - Maintenance of ocular viability is one of the major impediments to successful whole-eye transplantation. This review provides a comprehensive understanding of the current literature to help guide future studies in order to overcome this hurdle. A systematic multistage review of published literature was performed. Three specific questions were addressed: (1) Is recovery of visual function following eye transplantation greater in cold-blooded vertebrates when compared with mammals? (2) Is outer retina function following enucleation and reperfusion improved compared with enucleation alone? (3) Following optic-nerve transection, is there a correlation between retinal ganglion cell (RGC) survival and either time after transection or proximity of the transection to the globe? In a majority of the studies performed in the literature, recovery of visual function can occur after whole-eye transplantation in cold-blooded vertebrates. Following enucleation (and reperfusion), outer retinal function is maintained from 4 to 9 h. RGC survival following optic-nerve transection is inversely related to both the time since transection and the proximity of transection to the globe. Lastly, neurotrophins can increase RGC survival following optic-nerve transection. This review of the literature suggests that the use of a donor eye is feasible for whole-eye transplantation. PMID- 19286687 TI - GSTM1 and GSTT1 deletion genotypes in various spontaneous optic neuropathies in Arabs. AB - AIM: To investigate whether the prevalence GSTT1 and GSTM1 deletion genotypes (T0M1, T1M0 and T0M0) are increased in certain spontaneous optic neuropathies. METHODS: We compared the prevalence of GSTT1 and GSTM1 deletion genotypes in 108 Arab patients with optic neuritis (ON, 26 patients), LHON-like optic neuropathy (LLON, 35 patients), sporadic bilateral optic neuropathy in children (SBON, 21 patients) and non-arteritic ischaemic optic neuropathy (NAION, 26 patients) to 120 ethnicity-matched controls. Genotypes were determined by multiplex polymerase chain reaction. RESULTS: All three GST deletion genotypes were significantly more prevalent in the entire optic neuropathy group than in controls. When patients were stratified by optic neuropathy type, the prevalence of at least one deletion genotype was significantly increased in each type of optic neuropathy. CONCLUSIONS: These results imply that GST malfunction in the setting of GST deletion genotypes may interfere with metabolism of oxidative intermediates and may exacerbate direct or indirect pathological effects of oxidative stress on the optic nerve in the setting of these spontaneous optic neuropathies. It is possible that these GST polymorphisms are risk factors for the types of optic neuropathies investigated here. PMID- 19286688 TI - Alzheimer's disease and glaucoma: is there a causal relationship? AB - Evidence of a link between Alzheimer's disease (AD) and glaucoma has emerged from studies showing that patients with AD may have a significantly increased rate of glaucoma occurrence. In addition, it has been reported that patients with AD exhibit optic nerve degeneration and loss of retinal ganglion cells. In spite of intensive research, the clinical and genetic relationships between AD and glaucoma remain obscure. It is unclear whether the clinical correlation between the two diseases might be due to shared risk factors or the influence of one disorder on the other. Interestingly, certain observations may provide a clue towards a better understanding of the high rate of comorbidity reported between AD and glaucoma. In this article, we hypothesise that there may be a causal relationship between AD and glaucoma that may be explained by decreased cerebrospinal fluid pressure (CSFP) in patients with AD. A very recent study reported the intriguing new observation that mean CSFP was 33% lower in subjects with primary open-angle glaucoma than that of non-glaucomatous controls. It was noted that this observation supports the concept that an abnormal high trans lamina cribrosa pressure difference, whether the result of elevated intraocular pressure, reduced CSFP, or both, plays an important role in glaucomatous optic nerve damage. Interestingly, it was also reported that a substantial proportion of AD patients have very low CSFP. Therefore, we hypothesise that an abnormal high trans-lamina cribrosa pressure difference may explain why patients with AD have a greater risk for developing glaucoma. PMID- 19286689 TI - Primary birthing attendants and birth outcomes in remote Inuit communities--a natural "experiment" in Nunavik, Canada. AB - BACKGROUND: There is a lack of data on the safety of midwife-led maternity care in remote or indigenous communities. In a de facto natural "experiment", birth outcomes were assessed by primary birthing attendant in two sets of remote Inuit communities. METHODS: A geocoding-based retrospective birth cohort study in 14 Inuit communities of Nunavik, Canada, 1989-2000: primary birth attendants were Inuit midwives in the Hudson Bay (1529 Inuit births) vs western physicians in Ungava Bay communities (1197 Inuit births). The primary outcome was perinatal death. Secondary outcomes included stillbirth, neonatal death, post-neonatal death, preterm, small-for-gestational-age and low birthweight birth. Multilevel logistic regression was used to obtain the adjusted odds ratios (aOR) controlling for maternal age, marital status, parity, education, infant sex and plurality, community size and community-level random effects. RESULTS: The aORs (95% confidence interval) for perinatal death comparing the Hudson Bay vs Ungava Bay communities were 1.29 (0.63 to 2.64) for all Inuit births and 1.13 (0.48 to 2.47) for Inuit births at > or =28 weeks of gestation. There were no statistically significant differences in the crude or adjusted risks of any of the outcomes examined. CONCLUSION: Risks of perinatal death were somewhat but not significantly higher in the Hudson Bay communities with midwife-led maternity care compared with the Ungava Bay communities with physician-led maternity care. These findings are inconclusive, although the results excluding extremely preterm births are more reassuring concerning the safety of midwife-led maternity care in remote indigenous communities. PMID- 19286690 TI - Work-based social networks and health status among Japanese employees. AB - BACKGROUND: Despite the worldwide trend towards more time being spent at work by employed people, few studies have examined the independent influences of work based versus home-based social networks on employees' health. We examined the association between work-based social networks and health status by controlling for home-based social networks in a cross-sectional study. METHODS: By employing a two-stage stratified random sampling procedure, 1105 employees were identified from 46 companies in Okayama, Japan, in 2007. Work-based social networks were assessed by asking the number of co-workers whom they consult with ease on personal issues. The outcome was self-rated health; the adjusted OR for poor health compared employees with no network with those who have larger networks. RESULTS: Although a clear (and inverse) dose-response relationship was found between the size of work-based social networks and poor health (OR 1.53, 95% CI 1.03 to 2.27, comparing those with the lowest versus highest level of social network), the association was attenuated to statistical non-significance after we controlled for the size of home-based social networks. In further analyses stratified on age groups, in older workers (> or =50 years) work-based social networks were apparently associated with better health status, whereas home-based networks were not. The reverse was true among middle-aged workers (30-49 years). No associations were found among younger workers (<30 years). CONCLUSIONS: The present study suggests a differential association of alternative sources of social support on health according to age groups. We hypothesise that these patterns reflect generational differences in workers' commitment to their workplace. PMID- 19286691 TI - A population-based assessment of the familial component of acute kidney allograft rejection. AB - BACKGROUND: The genetic determinants of acute kidney transplant rejection (AR) are not well studied, and familial aggregation has never been demonstrated. The goal of this retrospective case-control study was to exploit the unique nature of the Utah Population Database (UPDB) to evaluate if AR or rejection-free survival aggregates in families. METHODS: We identified 891 recipients with genealogy data in the UPDB with at least one year of follow-up, of which 145 (16.1%) had AR and 77 recipients had biopsy-proven rejection graded >or=1A. We compared the genealogical index of familiality (GIF) in cases and controls (i.e. recipients with random assignment of rejection status). RESULTS: We did not find evidence for familial clustering of AR in the entire patient population or in the subgroup with early rejection (n = 52). When the subgroup of recipients with rejection grade >or=1A (n = 77) was analysed separately, we observed increased familial clustering (GIF = 3.02) compared to controls (GIF = 1.96), although the p-value did not reach the level of statistical significance (p = 0.17). Furthermore, we observed an increase in familial clustering in recipients who had a rejection free course (GIF = 2.45) as compared to controls (GIF = 2.08, p = 0.04). When all recipients were compared to non-transplant controls, they demonstrated a much greater degree of familiality (GIF = 2.03 versus GIF 0.63, p < 0.001). CONCLUSIONS: There is a familial component to rejection-free transplant course and trend to familial aggregation in recipients with AR grade 1A or higher. If a genetic association study is performed, there are families in Utah identified in the current study that can be targeted to increase the power of the test. PMID- 19286692 TI - Do patients with uric acid stones exhibit abnormal circadian blood pressure--a hypothesis. PMID- 19286693 TI - Restoration of hand function in a rat model of repair of brachial plexus injury. AB - The incurability of spinal cord injury and subcortical strokes is due to the inability of nerve fibres to regenerate. One of the clearest clinical situations where failure of regeneration leads to a permanent functional deficit is avulsion of the brachial plexus. In current practice, surgical re-implantation of avulsed spinal roots provides a degree of motor recovery, but the patients neither recover sensation nor the use of the hand. In the present rat study, we show that transplantation of cultured adult olfactory ensheathing cells restores the sensory input needed for a complex, goal-directed fore-paw function and re establishes synaptic transmission to the spinal grey matter and cuneate nucleus by providing a bridge for regeneration of severed dorsal root fibres into the spinal cord. Success in a first application of human olfactory ensheathing cells in clinical brachial plexus injury would open the way to the wider field of brain and spinal cord injuries. PMID- 19286694 TI - Role of subdural electrocorticography in prediction of long-term seizure outcome in epilepsy surgery. AB - Since prediction of long-term seizure outcome using preoperative diagnostic modalities remains suboptimal in epilepsy surgery, we evaluated whether interictal spike frequency measures obtained from extraoperative subdural electrocorticography (ECoG) recording could predict long-term seizure outcome. This study included 61 young patients (age 0.4-23.0 years), who underwent extraoperative ECoG recording prior to cortical resection for alleviation of uncontrolled focal seizures. Patient age, frequency of preoperative seizures, neuroimaging findings, ictal and interictal ECoG measures were preoperatively obtained. The seizure outcome was prospectively measured [follow-up period: 2.5 6.4 years (mean 4.6 years)]. Univariate and multivariate logistic regression analyses determined how well preoperative demographic and diagnostic measures predicted long-term seizure outcome. Following the initial cortical resection, Engel Class I, II, III and IV outcomes were noted in 35, 6, 12 and 7 patients, respectively. One child died due to disseminated intravascular coagulation associated with pseudomonas sepsis 2 days after surgery. Univariate regression analyses revealed that incomplete removal of seizure onset zone, higher interictal spike-frequency in the preserved cortex and incomplete removal of cortical abnormalities on neuroimaging were associated with a greater risk of failing to obtain Class I outcome. Multivariate logistic regression analysis revealed that incomplete removal of seizure onset zone was the only independent predictor of failure to obtain Class I outcome. The goodness of regression model fit and the predictive ability of regression model were greatest in the full regression model incorporating both ictal and interictal measures [R(2) 0.44; Area under the receiver operating characteristic (ROC) curve: 0.81], slightly smaller in the reduced model incorporating ictal but not interictal measures (R(2) 0.40; Area under the ROC curve: 0.79) and slightly smaller again in the reduced model incorporating interictal but not ictal measures (R(2) 0.27; Area under the ROC curve: 0.77). Seizure onset zone and interictal spike frequency measures on subdural ECoG recording may both be useful in predicting the long term seizure outcome of epilepsy surgery. Yet, the additive clinical impact of interictal spike frequency measures to predict long-term surgical outcome may be modest in the presence of ictal ECoG and neuroimaging data. PMID- 19286696 TI - Focal laser-lesions activate an endogenous population of neural stem/progenitor cells in the adult visual cortex. AB - CNS lesions stimulate adult neurogenic niches. Endogenous neural stem/progenitor cells represent a potential resource for CNS regeneration. Here, we investigate the response to unilateral focal laser-lesions applied to the visual cortex of juvenile rats. Within 3 days post-lesion, an ipsilateral increase of actively cycling cells was observed in cortical layer one and in the callosal white matter within the lesion penumbra. The cells expressed the neural stem/progenitor cell marker Nestin and the 473HD-epitope. Tissue prepared from the lesion area by micro-dissection generated self-renewing, multipotent neurospheres, while cells from the contralateral visual cortex did not. The newly formed neural stem/progenitor cells in the lesion zone might support neurogenesis, as suggested by the expression of Pax6 and Doublecortin, a marker of newborn neurons. We propose that focal laser-lesions may induce the emergence of stem/progenitor cells with neurogenic potential. This could underlie the beneficial effects of laser application in neurosurgery. PMID- 19286695 TI - Glucocerebrosidase mutations in clinical and pathologically proven Parkinson's disease. AB - Mutations in the glucocerebrosidase gene (GBA) are associated with Gaucher's disease, the most common lysosomal storage disorder. Parkinsonism is an established feature of Gaucher's disease and an increased frequency of mutations in GBA has been reported in several different ethnic series with sporadic Parkinson's disease. In this study, we evaluated the frequency of GBA mutations in British patients affected by Parkinson's disease. We utilized the DNA of 790 patients and 257 controls, matched for age and ethnicity, to screen for mutations within the GBA gene. Clinical data on all identified GBA mutation carriers was reviewed and analysed. Additionally, in all cases where brain material was available, a neuropathological evaluation was performed and compared to sporadic Parkinson's disease without GBA mutations. The frequency of GBA mutations among the British patients (33/790 = 4.18%) was significantly higher (P = 0.01; odds ratio = 3.7; 95% confidence interval = 1.12-12.14) when compared to the control group (3/257 = 1.17%). Fourteen different GBA mutations were identified, including three previously undescribed mutations, K7E, D443N and G193E. Pathological examination revealed widespread and abundant alpha-synuclein pathology in all 17 GBA mutation carriers, which were graded as Braak stage of 5 6, and had McKeith's limbic or diffuse neocortical Lewy body-type pathology. Diffuse neocortical Lewy body-type pathology tended to occur more frequently in the group with GBA mutations compared to matched Parkinson's disease controls. Clinical features comprised an early onset of the disease, the presence of hallucinations in 45% (14/31) and symptoms of cognitive decline or dementia in 48% (15/31) of patients. This study demonstrates that GBA mutations are found in British subjects at a higher frequency than any other known Parkinson's disease gene. This is the largest study to date on a non-Jewish patient sample with a detailed genotype/phenotype/pathological analyses which strengthens the hypothesis that GBA mutations represent a significant risk factor for the development of Parkinson's disease and suggest that to date, this is the most common genetic factor identified for the disease. PMID- 19286697 TI - Expression of endothelial protein C receptor in cortical peritubular capillaries associates with a poor clinical response in lupus nephritis. AB - OBJECTIVE: To study the membrane expression of endothelial protein C receptor (mEPCR) in the renal microvasculature in lupus nephritis (LN) as a potential marker of injury and/or prognostic indicator for response to therapy. METHODS: mEPCR expression was analysed by immunohistochemistry in normal kidney and in 59 biopsies from 49 patients with LN. Clinical parameters were assessed at baseline, 6 months and 1 year. RESULTS: mEPCR was expressed in the medulla, arterial endothelium and cortical peritubular capillaries (PTCs) in all biopsies with LN but not in the cortical PTCs of normal kidney. Positive mEPCR staining in >25% of the PTCs was observed in 16/59 biopsies and associated with poor response to therapy. Eleven (84.6%) of 13 patients with positive staining for mEPCR in >25% of the PTCs and follow-up at 6 months did not respond to therapy, compared with 8/28 (28.6%) with mEPCR staining in < or =25% PTCs, P = 0.0018. At 1 year, 10 (83.3%) of 12 patients with positive mEPCR staining in >25% of the PTCs did not respond to therapy (with two progressing to end-stage renal disease) compared with 8/24 (33.3%) with positive staining in < or =25% of the PTCs, P = 0.0116. Although tubulo-interstitial damage (TID) was always accompanied by mEPCR, this endothelial marker was extensively expressed in the absence of TID suggesting that poor response could not be attributed solely to increased TID. mEPCR expression was independent of International Society of Nephrology/Renal Pathology Society class, activity and chronicity indices. CONCLUSION: Increased mEPCR expression in PTCs may represent a novel marker of poor response to therapy for LN. PMID- 19286699 TI - Abstracts of Rheumatology '09. April 28-May 1, 2009. Glasgow, Scotland. PMID- 19286698 TI - Association between depression and coronary artery calcification in women with systemic lupus erythematosus. AB - OBJECTIVES: To determine the associations between depression, cardiovascular risk factors and coronary artery calcification (CAC) in women with SLE and controls. METHODS: CAC was measured using electron-beam CT (EBCT). Traditional, inflammatory and lupus-related risk factors as well as depressive symptoms (Center for Epidemiologic Studies Depression Scale-CES-D) were measured at a single study visit in 161 women with SLE and 161 age- and race frequency-matched female healthy controls. RESULTS: Women with SLE reported more depressive symptoms than controls, with 27% of SLE and 15% of controls having CES-D scores suggestive of clinical depression. SLE women were more likely to have CAC, as well as more severe CAC compared with controls. Among the SLE women, depression was associated with greater than 2-fold odds of having any CAC [odds ratio (OR) 2.48; 95% CI 1.05, 5.87; P = 0.04], independent of traditional risk factors (age, hypertension and triglycerides) and inflammatory markers. However, when BMI was included among the covariates, the association between depression and CAC was attenuated, indicating the potential mediating role of BMI. Depression was not a risk factor for CAC in controls. CONCLUSIONS: In women with SLE, depression was associated with CAC. This association was mediated by BMI. Depression and adiposity may add to the inflammatory burden of SLE, thus contributing to cardiovascular disease risk. PMID- 19286741 TI - Classification of pain in cancer patients--a systematic literature review. AB - One of the aims of the European Palliative Care Research Collaborative (EPCRC) is to achieve consensus on a classification system for cancer pain. We performed a systematic literature review to identify existing classification systems and domains/items used to classify cancer patients with pain. In a systematic search in the databases Medline and Embase, covering 1986-2006, 692 hits were obtained. 92 papers were evaluated to address pain classification. Six standardised classification systems were identified; three of them systematically developed and partially validated. Both pain characteristics and patient characteristics relevant for cancer pain classification were included in the classification systems. All but one of the standardised systems aim at predicting treatment response or adequacy of treatment. Several domains and items used to describe cancer pain but not formally described as part of a classification system were also identified and systematized. The existing approaches to pain classification in cancer patients are different, mostly not thoroughly validated, and none is widely applied. An internationally accepted classification system for cancer pain could improve research and cancer pain management. This systematic review suggests a need for developing an international consensus on how to classify pain in cancer patients. PMID- 19286742 TI - The neural basis of effective memory therapy in a patient with limbic encephalitis. AB - BACKGROUND: An fMRI study is described in which a postencephalitic woman with amnesia used a wearable camera which takes photographs passively, without user intervention, to record and review recent autobiographical events. "SenseCam" generates hundreds of images which can subsequently be reviewed quickly or one by one. RESULTS: Memory for a significant event was improved substantially when tested after 4.5 weeks, if the patient viewed SenseCam images of the event every 2 days for 3 weeks. In contrast, after only 3.5 weeks, her memory was at chance levels for a similarly significant event which was reviewed equally often, but using a written diary. During the fMRI scan, the patient viewed images of these two events, plus images of an unrehearsed event and images from a novel "control" event that she had never experienced. There was no difference in behavioural responses or in activation when the unrehearsed and novel conditions were compared. Relative to the written-rehearsed condition, successful recognition of the images in the SenseCam-rehearsed condition was associated with activation of frontal and posterior cortical regions associated with normal episodic memory. CONCLUSION: SenseCam images may provide powerful cues that trigger the recall and consolidation of stored but inaccessible memories. PMID- 19286745 TI - Number of global clinical trials done in UK fell by two thirds after EU directive. PMID- 19286746 TI - Comprehensive care and HIV prophylaxis after sexual assault in rural South Africa: the Refentse intervention study. AB - PROBLEM: Although international guidelines specify the central role of the health sector in providing comprehensive care, including HIV post-exposure prophylaxis (PEP), after sexual assault, in both industrialised and developing countries there are many challenges to providing timely and comprehensive services. DESIGN: A nurse driven model of post-rape care was integrated into existing hospital services; the before and after study design evaluated impacts on quality of care, reviewing 334 hospital charts and conducting interviews with 16 service providers and 109 patients. SETTING: 450 bed district hospital in rural South Africa. KEY MEASURES FOR IMPROVEMENT: Quality of care after rape (forensic history and examination, provision of emergency contraception, prophylaxis for sexually transmitted infections, referrals); provision of HIV counselling and testing and provision and completion of full 28 day course of PEP; and service utilisation (number of service providers seen on first visit and number of rape cases presenting to hospital per month). STRATEGIES FOR CHANGE: After completing baseline research, we introduced a five part intervention model, consisting of a sexual violence advisory committee, hospital rape management policy, training workshop for service providers, designated examining room, and community awareness campaigns. Effect of change Existing services were fragmented and of poor quality. After the intervention, there were considerable improvements in clinical history and examination, pregnancy testing, emergency contraception, prophylaxis for sexually transmitted infections; HIV counselling and testing, PEP, trauma counselling, and referrals. Completion of the 28 day course of PEP drugs increased from 20% to 58%. LESSONS LEARNT: It is possible to improve the quality of care after sexual assault, including HIV prophylaxis, within a rural South African hospital at modest cost, using existing staff. With additional training, nurses can become the primary providers of this care. PMID- 19286747 TI - Investigating the thyroid nodule. PMID- 19286748 TI - Sex differences in child and adolescent mortality in the Nordic countries, 1981- 2000. AB - AIMS: Unlike the situation for infant and adult mortality, there are only a few studies on child and adolescent mortality. The purpose of this study was to examine sex differences in child and adolescent mortality by age and cohort in four Nordic countries over a 20-year period. METHODS: Data on all live-born children were received from national population-covering birth registries from 1981 to 2000 (Denmark, n = 1,184,926; Finland, n = 841,470 (from 1987 to 2000); Norway, n = 1,090,127; and Sweden, n = 1,961,911). Data on mortality and causes of death until the age of 20 years were received from the national cause-of-death registers. RESULTS: The overall mortality rates were higher in Denmark and Norway than in Finland and Sweden, among both boys and girls, and the difference between countries increased over time. In all countries, boys had higher mortality rates than girls. Overall, the sex difference was larger in Denmark and Norway (36% and 33% higher, respectively) than in Finland and Sweden (both 24%). The sex differences were more pronounced for deaths of those aged 5-19 years than for those aged 0-4 years. Twelve per cent of all deaths among boys and 10% of those among girls were due to external causes, mainly unintentional injury deaths or intentional deaths. For children and adolescents aged 5-19 years, the corresponding figures were 43% for boys and 35% for girls. CONCLUSIONS: Boys have excess mortality, but the sex difference is lower in countries with lower mortality. Boys' excess mortality was only partly accounted for by deaths from external causes. Avoidable deaths and causes of death need further investigation. PMID- 19286749 TI - Clerics die, doctors survive: a note on death risks among highly educated professionals. AB - AIMS: Mortality is strongly associated with education. We present relative death risks of men and women in 12 educational/ occupational groups in Sweden today, with a focus on individuals with higher education. METHODS: Results from Cox regressions are reported for 12 educational groups with special emphasis on those with professional education, e.g. clerics, physicians, people with medical PhDs, and university teachers. The study is based on register data of the total Swedish population in the age group of 30-64 (n = 3,734,660). RESULTS: There is a considerable variation in mortality between educational groups. Men with compulsory education run a risk that is more than three times higher than that of professors outside medicine, and other educational groups fall in between. Medical doctors and physicians have relatively low death risks compared to those with compulsory education - less than 50% among men and less than 60% among women - although professors in medicine deviate by having higher risks than their colleagues in other subjects. Those with a theological exam show higher risks of dying during the follow-up period compared to others of a similar educational level. Professors outside medicine experience the lowest death risks of all identified groups. CONCLUSIONS: Men and women with a professional education have comparatively low death risks, particularly low among medical doctors and university employees, while the clergy seems to experience relatively higher death risks than others with a similar level of education. These patterns may reflect the effects of education as well as the selection of men and women to higher education. PMID- 19286750 TI - Direct observation and completion of treatment of tuberculosis in rural areas of China. AB - AIMS: To analyse the current status of directly observed therapy (DOT) and completion of treatment of tuberculosis (TB) in two rural areas of China. METHODS: Two rural counties with low DOT rates were deliberately selected as study sites. Face-to-face interviews were conducted by trained investigators with a structured questionnaire to investigate the characteristics of patients and the TB service that they had received. The associations between treatment completion and potential factors were estimated by computing odds ratios (ORs), as well as their 95% confidence intervals (CIs), from an unconditional logistic regression model. RESULTS: Among 601 patients, 2.2% were treated with direct observation by health workers, 6.2% were supervised by family members, and 91.7% were treated with self-administered therapy. The treatment completion rate was found to be significantly associated with sputum smear test and adverse reaction to anti tuberculosis drugs, but not with direct observation by health workers (OR 1.81, 95% CI 0.23-14.38) or by family members (OR 1.14, 95% CI 0.38-3.41). Frequent home visiting by health workers (!1 visit/month) could help to increase the completion rate (OR 3.15, 95% CI 1.30-7.63). CONCLUSIONS: No significant difference was found in the rate of completion of TB treatment between direct observation and self-supervision groups in two rural areas with lower DOT coverage. How to build a feasible DOT strategy that is accepted by both patients and healthcare providers needs to be considered by policy-makers. Other elements apart from DOT are necessary to ensure a successful TB programme. PMID- 19286751 TI - Perceived change in life satisfaction following epilepsy diagnosis. AB - AIMS: To study the development in perceived life satisfaction of people with epilepsy. METHODS: In 2005, members of the Norwegian Epilepsy Association responded to a version of the Self-Anchoring Striving Scale (termed Cantril's Ladder). In this article, we compare the answers given to two questions, one related to the person's life satisfaction today, and the other related to the person's life satisfaction 5 years ago. The same people were asked about time since epilepsy onset. Those who reported being diagnosed during the previous 5 years were compared with those who had been diagnosed earlier, with regard to reported life satisfaction, at the present and 5 years ago. The hypothesis to be tested was that people who have been diagnosed with epilepsy report their life satisfaction as being reduced in comparison to their life satisfaction before the diagnosis. RESULTS: The hypothesis was confirmed. The respondents in our study who had been diagnosed less than 5 years ago perceived their life satisfaction to be reduced by almost a standard deviation as compared to their life satisfaction before the diagnosis. CONCLUSIONS: People diagnosed with epilepsy perceive their life satisfaction to be reduced as a consequence of the diagnosis, and should be supported in their coping with the disease. This conclusion should be drawn with caution, owing to a low response rate and difficulties related to retrospective self-reports. Prospective population-based studies are needed to explore the causal pathways. PMID- 19286752 TI - Review: renal protection by inhibition of the renin-angiotensin-aldosterone system. AB - Renin-angiotensin-aldosterone system (RAAS) inhibition exerts a renoprotective effect independent of blood pressure reduction. Many studies using an end-point of proteinuria compared the effects of angiotensin-converting enzyme inhibitor (ACE-I) or angiotensin receptor blocker (ARB) monotherapy with combination ACE I/ARB therapy. Despite methodological limitations, most studies suggest that combination therapy provides a greater antiproteinuric effect than monotherapy, perhaps because of more prolonged and complete RAAS inhibition. COOPERATE and ONTARGET used more robust end-points to study renoprotective effects. In COOPERATE, combination therapy resulted in significantly longer times to doubling serum creatinine or developing end-stage renal disease than trandolapril or losartan monotherapy. However, a secondary ONTARGET finding was that combination therapy significantly increased the risk for renal dysfunction compared with ramipril or telmisartan alone. Eventually, the VA NEPHRON-D trial should provide definitive data relating to patients with diabetic nephropathy. RESULTS: of AVOID suggest the renoprotective benefits of combination therapy extend to the direct renin inhibitors (DRI). In AVOID, combination therapy with aliskiren, a DRI, and losartan resulted in 20% greater protein excretion decrement than losartan monotherapy. Future trials should examine higher RAAS inhibitor doses, facilitate differentiation of renoprotective and antihypertensive effects of RAAS blockade, and use end-points that robustly demonstrate renoprotective effects. PMID- 19286753 TI - Role of oxidative stress in the natriuresis induced by central administration of angiotensin II. AB - INTRODUCTION: Central administration of angiotensin II (Ang II) is known to reduce urinary volume and to increase sodium and potassium excretion. Recently, a novel signalling mechanism for Ang II in the periphery has been shown to involve reduced nicotinamide adenine dinucleotide phosphate [NAD(P)H] oxidase-derived reactive oxygen species (ROS).Although ROS are now known to be involved in numerous Ang II-regulated processes in peripheral tissues, and are increasingly implicated in CNS neurodegenerative diseases, the role of ROS in central regulation of Ang II-induced hydromineral metabolism remains unexplored.The hypothesis that ROS are involved in central Ang II signalling and in Ang II dependent antidiuresis, natriuresis and kaliuresis was tested by the use of selective antagonists of the NAD(P)H oxidase cascade. MATERIALS AND METHODS: In intracerebroventricular (ICV)-cannulated rats,Ang II was injected ICV and urinary sodium and potassium excretion was assessed at 1-, 3-, and 6-hour periods of urine collection. Urine sample was analysed for sodium and potassium concentration using a flame photometer. The role of NAD(P)H oxidase-dependent signalling cascade was evaluated using the selective NAD(P)H oxidase inhibitor, apocynin; the superoxide dismutase mimetic, 4-hydroxy-2,2,6,6 tetramethylpiperidine-1-oxyl (tempol); and the protein kinase C inhibitor, chelerythrine. RESULTS: ICV administration of Ang II to conscious hydrated rats resulted in a significant decrease in urinary volume in the first hour, and an increased sodium and potassium excretion during the 6-hour period of urine collection, which was most effective during the 3 and 6 h. Interference with the NAD(P)H oxidase signalling by central administration of apocynin, tempol or chelerythrine, blunted the natriuretic and kaliuretic effect induced by central administration of Ang II, without affecting its antidiuretic action. CONCLUSION: This study demonstrates that increases of urinary sodium and potassium excretion elicited by central administration of Ang II are mediated by NAD(P)H oxidase- dependent production of superoxide and protein kinase C activation in conscious hydrated rats. PMID- 19286754 TI - Local renin-angiotensin system regulates left ventricular hypertrophy induced by swimming training independent of circulating renin: a pharmacological study. AB - INTRODUCTION: This study addressed the role of the local renin-angiotensin system (RAS) in the left ventricular hypertrophy (LVH) induced by swimming training using pharmacological blockade. MATERIALS AND METHODS: Female Wistar rats treated with enalapril maleate (60 mg.kg(-1).d( -1), n=38), losartan (20 mg.kg(-1).d(-1), n=36) or high salt diet (1% NaCl, n=38) were trained by two protocols (T1: 60-min swimming session, 5 days per week for 10 weeks and T2: the same T1 protocol until the 8(th) week, then 9(th) week they trained twice a day and 10(th) week they trained three times a day). Salt loading prevented activation of the systemic RAS. Haemodynamic parameters, soleus citrate synthase (SCS) activity and LVH (left ventricular/body weight ratio, mg/g) were evaluated. RESULTS: Resting heart rate decreased in all trained groups. SCS activity increased 41% and 106% in T1 andT2 groups, respectively. LVH was 20% and 30% in T1 andT2 groups, respectively. Enalapril prevented 39% of the LVH in T2 group (p<0.05). Losartan prevented 41% in T1 and 50% inT2 (p<0.05) of the LVH in trained groups. Plasma renin activity (PRA) was inhibited in all salt groups and it was increased in T2 group. CONCLUSIONS: These data provide evidence that the physiological LVH induced by swimming training is regulated by local RAS independent from the systemic, because the hypertrophic response was maintained even when PRA was inhibited by chronic salt loading. However, other systems can contribute to this process. PMID- 19286755 TI - Superoxide dismutase and catalase anti-oxidant activity in leucocyte lysates from hypertensive patients: effects of eprosartan treatment. AB - INTRODUCTION: In an earlier study, our group reported that circulating leucocytes in hypertensive (HT) patients show a significant increase in oxidative stress compared to the control group, and this normalised after two months of treatment with eprosartan.(1) It can be speculated that these facts may be attributable to a possible reduction in anti-oxidative activity in untreated HT patients, which would be corrected by eprosartan. MATERIALS AND METHODS: In this observational pilot study, superoxide dismutase and catalase activities were evaluated in leucocyte lysates in a group of 21 HT patients at baseline and after two months of treatment with eprosartan (600 mg/ day). For the control group, 25 normotensive volunteers were recruited with comparable characteristics to the patients. RESULTS: The results obtained indicate, paradoxically, that the untreated HT patients present greater anti-oxidant enzyme activity than the control group. CONCLUSION: This result could be interpreted as a cell defence mechanism against the greater oxidative stress that exists in these patients.This hypothesis is consistent with the facts reported previously by our group in which a reduction in oxidative stress was found after two months of treatment with eprosartan.( 1) Upon reducing this stress, less anti-oxidative activity would be necessary, just as was observed in the present study after two months of treatment with eprosartan. PMID- 19286756 TI - Meta-analysis of association of ACE2 G8790A polymorphism with Chinese Han essential hypertension. AB - INTRODUCTION: We performed a meta-analysis to assess the association of ACE2 G8790A polymorphism with essential hypertension. MATERIALS AND METHODS: The references were retrieved through MEDLINE, Wanfang and VIP Information.The analyses were performed by the software STATA 9.0. RESULTS: No significant publication bias was observed.The combined data showed no association of the frequencies of the A allele with essential hypertension odds ratio (OR)=1.09 95% confidence interval (CI)=0.98-1.20;p=0.11 in female patients. No associations were shown between the frequencies of the A allele with the genetic susceptibility to essential hypertension in male patients (OR=1.11 95% CI=0.95 1.29; p=0.19). CONCLUSION: The meta-analysis suggests that ACE2 G8790A polymorphism may not be a genetic risk factor for essential hypertension in a Chinese Han population. PMID- 19286757 TI - Association of the insertion/deletion polymorphism of the angiotensin-converting enzyme gene with type 2 diabetes in two ethnic groups of Jerba Island in Tunisia. AB - INTRODUCTION: The aim of the current study was to evaluate the role of angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism on the prediction of type 2 diabetes in two ethnic populations from Jerba Island,Tunisia. METHODS: In this study, we analysed the genotypic and the allelic distributions of the ACE I/D polymorphism and conducted a case/control association study between healthy normoglycaemic controls and diabetic patients in the two studied groups.ACE gene polymorphism was analysed by polymerase chain reaction in 272 individuals consisting of 172 diabetic subjects and 100 controls. RESULTS: The genotype frequencies for DD, ID and II were 75.50%, 19.60% and 4.89% inArabs and 76.66%, 16.66% and 6.67% in Berbers, respectively, in the case group, and 42.85%, 35.71% and 21.43% inArabs and 57.50%, 22.50% and 20.00% in Berbers, respectively, in the control group.The DD frequency was significantly higher in the case group than in the control group (p<0.001), suggesting that the DD genotype is associated with an increased susceptibility to type 2 diabetes in our study populations. CONCLUSIONS: The current investigation provides new evidence regarding the role of the ACE I/D polymorphism in the pathogenesis of type 2 diabetes in Jerbian populations. Furthermore, it underlines the importance of ethnicity, which should be considered in all studies aiming to test the genetic effects on the susceptibility to type 2 diabetes. PMID- 19286758 TI - Impact of renin-angiotensin system polymorphisms on renal haemodynamic responsiveness to acute angiotensin-converting enzyme inhibition in type 2 diabetes mellitus. AB - INTRODUCTION: The aim of this study was to document the impact of renin angiotensin system (RAS) polymorphisms on renal haemodynamics and renal hormones in type 2 diabetes mellitus. MATERIALS AND METHODS: Fifty-nine adult patients were studied. Renal haemodynamics were evaluated using 99mTc-MAG3 clearance (MAG3( Cle)) using Bubeck's method and captopril renogram. RAS hormones and angiotensin-converting enzyme (ACE) levels were measured before and after captopril.ACE, angiotensin II type 1 receptor and angiotensinogen gene polymorphisms were analysed. RESULTS: Post-captopril MAG3(Cle) values were significantly lower in patients with microalbuminuria compared to nonproteinuric patients. Statistically significant negative correlation was found between clearance percentage change values and HbA(1c) levels (r: -0.42, p=0.009). MAG3(Cle) was relatively lower following captopril administration in DD patients, while a relative increment was observed in I allele carriers (p=0.02).TheAC-CC group had significantly higher mean post-captopril clearance value compared to the AA genotype (480.9+/-56.1 ml/min/1.73 m(2) vs. 428.4+/-74.8 ml/min/1.73 m(2), p=0.022). CONCLUSIONS: Our data indicate that the heterogeneity of patients' response to ACE inhibition is, at least partly, genetically determined, and the genetic polymorphisms in RAS might predict the acute responsiveness to ACE inhibitors. PMID- 19286759 TI - Association between the angiotensin-converting enzyme (insertion/deletion) and angiotensin II type 1 receptor (A1166C) polymorphisms and breast cancer among Brazilian women. AB - INTRODUCTION: We evaluated the association between components of the renin angiotensin system and the development of breast cancer in a case-control study by means of angiotensin-converting enzyme (ACE) insertion/deletion (I/D) and angiotensin II type 1 (AT( 1))-receptor A1166C polymorphisms. METHODS: Genotyping was performed by PCR-RFLP (restriction fragment length polymorphism) or PCR (polymerase chain reaction) using genomic DNA extracted from buccal cells of subjects with (101 cases) or without (307 controls) breast cancer. RESULTS: The frequencies of genotypes for ACE were: DD, ID and II (in %: cases: 60; 20; 20; controls: 46; 37; 17; p=0.019, chi(2)); and for AT(1)receptor were:AA,AC and CC (in %: cases: 65; 30; 5; controls: 51; 44; 5; p=0.114, chi( 2)).The results suggested that the A1166C polymorphism was not associated with breast cancer risk. On the other hand, for the ACE (I/D), there seemed to be different risks for cancer between cases and controls. CONCLUSIONS: The ID genotype was less frequently associated with the disease than were the DD or II; that is, women with the ID genotype were 3.1 times less likely to develop breast cancer than those with the other genotypes.The ID genotype might be protective against breast cancer and the ACE (I/D) polymorphism a possible target for developing genetic markers for breast cancer. PMID- 19286760 TI - Bone-resorbing cells in multiple myeloma: osteoclasts, myeloma cell polykaryons, or both? AB - Myeloma bone disease (MBD) leads to progressive destruction of the skeleton and is the most severe cause of morbidity in multiple myeloma. Its pathogenetic mechanisms are not fully understood, though the current evidence points to osteoclast (OC) hyperactivity coupled with defective osteoblast function unable to counteract bone resorption. OCs are generated in bone marrow by myeloid progenitors through increased levels of receptor activator of nuclear factor kappaB ligand and M-CSF, whose intracellular pathways propagate signals that activate sequential transcription factors, resulting in the production of major OC enzymes that drive specific functions such as acidification and degradation of the bone matrix. Osteolytic lesions, however, are not characterized by massive OC content, whereas malignant plasma cells, which are usually present in a high number, may occur as large multinucleated cells. The possibility that myeloma cells fuse and generate polykaryons in vivo is suggested by the in vitro formation of multinuclear cells that express tartrate-resistant acid phosphatase and produce pits and erosive lacunae on experimental osteologic substrates. Further, the detection in vivo of polykaryons with chromosome translocations typical of myeloma cells lends support to the view that myeloma polykaryons may act as functional OCs and participate in the skeletal destruction by resorbing bone. PMID- 19286761 TI - Myeloma bone disease: recent advances in biology, diagnosis, and treatment. AB - Bone disease is a hallmark of multiple myeloma (MM). Occurring in the majority of MM patients, it is associated with bone pain, fractures, and hypercalcemia and has major impacts on quality of life. Furthermore, bone resorption activity has been shown to be an independent risk factor for overall survival in patients with symptomatic MM. Myeloma is characterized by a unique form of bone disease with lytic bone destruction that is not followed by reactive bone formation (uncoupling). This review focuses on recent advances in our understanding of the biology of osteoclast activation and osteoblast inhibition in MM, diagnostic standards, and recent progress in treatment options for myeloma bone disease. Translational research has enabled a rapid transfer of mechanistic insights from the bench to the bedside and will hopefully result in better treatment options and outcome for patients in near future. PMID- 19286762 TI - Thyroid storm after intensity-modulated radiation therapy: a case report and discussion. AB - A 43-year-old man with locally advanced squamous cell carcinoma of the base of tongue was treated with induction chemotherapy followed by intensity-modulated radiation therapy (IMRT). Within 20 days post-treatment, the patient developed clinical symptoms highly suggestive of hyperthyroidism. Two and one half months after completion of therapy, the patient developed severe thyrotoxicosis, which, in retrospect, appears to have met the criteria for thyroid storm. This case history illustrates a previously unreported, life-threatening complication of external-beam radiation that should be considered in patients receiving IMRT therapy involving the thyroid. Diagnosis of the patient's hyperthyroidism and probable thyroid storm was difficult to recognize because of the significant overlap between the signs and symptoms of severe thyrotoxicosis and the expected toxicities of his cancer therapy. PMID- 19286763 TI - Ghost writers in the sky. PMID- 19286764 TI - Association of duration of television viewing in early childhood with the subsequent development of asthma. AB - OBJECTIVE: To investigate whether duration of television (TV) viewing in young children is associated with subsequent development of asthma. METHODS: Children taking part in the Avon Longitudinal Study of Parents and Children (ALSPAC) with no wheeze up to the age of 3.5 years and follow-up data at 11.5 years of age took part in a prospective longitudinal cohort study. The main outcome measure was asthma, defined as doctor-diagnosed asthma by 7.5 years of age with symptoms and/or treatment in the previous 12 months at 11.5 years of age. Parental report of hours of TV viewing per day by the children was ascertained at 39 months. RESULTS: In children with no symptoms of wheeze at 3.5 years of age and follow-up data at 11.5 years of age, the prevalence of asthma was 6% (185/3065). Increased TV viewing at 3.5 years was associated with increased prevalence of asthma at 11.5 years of age (p for linear trend = 0.0003). Children who watched television for >2 h/day were almost twice as likely to develop asthma by 11.5 years of age as those watching TV for 1-2 h/day (adjusted odds ratio 1.8 (95% CI 1.2 to 2.6)). CONCLUSION: Longer duration of TV viewing in children with no symptoms of wheeze at 3.5 years of age was associated with the development of asthma in later childhood. PMID- 19286766 TI - Intra-articular bupivacaine: potentially chondrotoxic? PMID- 19286767 TI - Assessment of platelet inhibition secondary to clopidogrel and aspirin therapy in preoperative acute surgical patients measured by Thrombelastography Platelet Mapping. AB - BACKGROUND: Increasing numbers of patients prescribed clopidogrel and aspirin are presenting for non-elective surgery. No consensus on the timing of surgery exists after withdrawal of antiplatelet and tests of platelet function are not routinely available. The Thrombelastography Platelet Mapping (TEG-PM) assay is designed to assess platelet inhibition secondary to antiplatelet therapy. We assessed its ability to detect platelet inhibition in preoperative acute surgical patients. METHODS: We conducted a prospective observational study in three groups of preoperative patients: those taking clopidogrel or aspirin up to admission, and a control group. TEG-PM was performed on the day of admission and alternate days until surgery. RESULTS: Mean (SD) platelet thromboxane A(2) receptor inhibition in the control group was 17.5% (23.8) (n=20), 52.6% (32.3) (n=18) in the aspirin group, and 31.9% (27.6) (n=21) in the clopidogrel group (P<0.01). Mean (SD) platelet adenosine diphosphate (ADP) receptor inhibition in the control group was 47.8% (18.9) (n=20), 52.6% (19.7) (n=18) in the aspirin group, and 71.5% (18.4) (n=21) in the clopidogrel group (P<0.01). Among the clopidogrel group awaiting surgery, mean platelet ADP channel inhibition decreased on day 3 to 67.1% (24.7) (n=11), 48.8% (24.4) (n=4) on day 5, and 36.1% (15.9) (n=2) on day 7 (P=0.57). CONCLUSIONS: TEG-PM can identify statistically significant platelet inhibition after antiplatelet therapy; however, the overlap in platelet receptor inhibition between the three groups is likely to limit the clinical usefulness of this test. PMID- 19286768 TI - Intraoperative cardiac arrest in acquired long QT syndrome. AB - A healthy female sustained a life-threatening arrhythmia and cardiac arrest while undergoing routine surgery under general anaesthesia. Resuscitation was prolonged but successful, with a complete neurological recovery. PMID- 19286769 TI - Real-time visualization of ultrasound-guided retrobulbar blockade: an imaging study. PMID- 19286770 TI - Is a decrease in capillary density dangerous? PMID- 19286771 TI - Convulsions associated with ropivacaine 300 mg for brachial plexus block. PMID- 19286772 TI - Urinary catheterization in labour. PMID- 19286773 TI - Tracheal intubation using an AirWay Scope in a patient with Halo-Vest Fixation for upper cervical spine injury. PMID- 19286774 TI - Vallecular cyst causing a difficult intubation. PMID- 19286775 TI - Weight loss and 2,4-dinitrophenol poisoning. PMID- 19286776 TI - Interaction between clonidine and escitalopram. PMID- 19286777 TI - Tilting at windmills? PMID- 19286778 TI - De novo biosynthesis of vanillin in fission yeast (Schizosaccharomyces pombe) and baker's yeast (Saccharomyces cerevisiae). AB - Vanillin is one of the world's most important flavor compounds, with a global market of 180 million dollars. Natural vanillin is derived from the cured seed pods of the vanilla orchid (Vanilla planifolia), but most of the world's vanillin is synthesized from petrochemicals or wood pulp lignins. We have established a true de novo biosynthetic pathway for vanillin production from glucose in Schizosaccharomyces pombe, also known as fission yeast or African beer yeast, as well as in baker's yeast, Saccharomyces cerevisiae. Productivities were 65 and 45 mg/liter, after introduction of three and four heterologous genes, respectively. The engineered pathways involve incorporation of 3-dehydroshikimate dehydratase from the dung mold Podospora pauciseta, an aromatic carboxylic acid reductase (ACAR) from a bacterium of the Nocardia genus, and an O-methyltransferase from Homo sapiens. In S. cerevisiae, the ACAR enzyme required activation by phosphopantetheinylation, and this was achieved by coexpression of a Corynebacterium glutamicum phosphopantetheinyl transferase. Prevention of reduction of vanillin to vanillyl alcohol was achieved by knockout of the host alcohol dehydrogenase ADH6. In S. pombe, the biosynthesis was further improved by introduction of an Arabidopsis thaliana family 1 UDP-glycosyltransferase, converting vanillin into vanillin beta-D-glucoside, which is not toxic to the yeast cells and thus may be accumulated in larger amounts. These de novo pathways represent the first examples of one-cell microbial generation of these valuable compounds from glucose. S. pombe yeast has not previously been metabolically engineered to produce any valuable, industrially scalable, white biotech commodity. PMID- 19286779 TI - Infection of Melanoplus sanguinipes grasshoppers following ingestion of rangeland plant species harboring vesicular stomatitis virus. AB - Knowledge of the many mechanisms of vesicular stomatitis virus (VSV) transmission is critical for understanding of the epidemiology of sporadic disease outbreaks in the western United States. Migratory grasshoppers [Melanoplus sanguinipes (Fabricius)] have been implicated as reservoirs and mechanical vectors of VSV. The grasshopper-cattle-grasshopper transmission cycle is based on the assumptions that (i) virus shed from clinically infected animals would contaminate pasture plants and remain infectious on plant surfaces and (ii) grasshoppers would become infected by eating the virus-contaminated plants. Our objectives were to determine the stability of VSV on common plant species of U.S. Northern Plains rangelands and to assess the potential of these plant species as a source of virus for grasshoppers. Fourteen plant species were exposed to VSV and assayed for infectious virus over time (0 to 24 h). The frequency of viable virus recovery at 24 h postexposure was as high as 73%. The two most common plant species in Northern Plains rangelands (western wheatgrass [Pascopyrum smithii] and needle and thread [Hesperostipa comata]) were fed to groups of grasshoppers. At 3 weeks postfeeding, the grasshopper infection rate was 44 to 50%. Exposure of VSV to a commonly used grasshopper pesticide resulted in complete viral inactivation. This is the first report demonstrating the stability of VSV on rangeland plant surfaces, and it suggests that a significant window of opportunity exists for grasshoppers to ingest VSV from contaminated plants. The use of grasshopper pesticides on pastures would decrease the incidence of a virus amplifying mechanical vector and might also decontaminate pastures, thereby decreasing the inter- and intraherd spread of VSV. PMID- 19286780 TI - Fusobacterium nucleatum ATCC 10953 requires Actinomyces naeslundii ATCC 43146 for growth on saliva in a three-species community that includes Streptococcus oralis 34. AB - Formation of dental plaque is a developmental process involving initial and late colonizing species that form polymicrobial communities. Fusobacteria are the most numerous gram-negative bacteria in dental plaque, but they become prevalent after the initial commensal colonizers, such as streptococci and actinomyces, have established communities. The unusual ability of these bacteria to coaggregate with commensals, as well as pathogenic late colonizers, has been proposed to facilitate colonization by the latter organisms. We investigated the integration of Fusobacterium nucleatum into multispecies communities by employing two in vitro models with saliva as the sole nutritional source. In flow cell biofilms, numbers of cells were quantified using fluorescently conjugated antibodies against each species, and static biofilms were analyzed by quantitative real-time PCR (q-PCR) using species-specific primers. Unable to grow as single-species biofilms, F. nucleatum grew in two-species biofilms with Actinomyces naeslundii but not with Streptococcus oralis. However, enhanced growth of fusobacteria was observed in three-species biofilms, indicating that there was multispecies cooperation. Importantly, these community dynamics yielded an 18-fold increase in the F. nucleatum biomass between 4 h and 18 h in the flow cell inoculated with three species. q-PCR analysis of static biofilms revealed that maximum growth of the three species occurred at 24 h to 36 h. Lower numbers of cells were observed at 48 h, suggesting that saliva could not support higher cell densities as the sole nutrient. Integration of F. nucleatum into multispecies commensal communities was evident from the interdigitation of fusobacteria in coaggregates with A. naeslundii and S. oralis and from the improved growth of fusobacteria, which was dependent on the presence of A. naeslundii. PMID- 19286781 TI - Molecular epidemiology and characterization of Campylobacter spp. isolated from wild bird populations in northern England. AB - Campylobacter infections have been reported at prevalences ranging from 2 to 50% in a range of wild bird species, although there have been few studies that have investigated the molecular epidemiology of Campylobacter spp. Consequently, whether wild birds are a source of infection in humans or domestic livestock or are mainly recipients of domestic animal strains and whether separate cycles of infection occur remain unknown. To address these questions, serial cross sectional surveys of wild bird populations in northern England were carried out over a 2-year period. Fecal samples were collected from 2,084 wild bird individuals and screened for the presence of Campylobacter spp. A total of 56 isolates were recovered from 29 birds sampled at 15 of 167 diverse locales. Campylobacter jejuni, Campylobacter lari, and Campylobacter coli were detected by PCR, and the prevalences of different Campylobacter spp. in different avian families ranged from 0% to 33%. Characterization of 36 C. jejuni isolates by multilocus sequence typing revealed that wild birds carry both livestock associated and unique strains of C. jejuni. However, the apparent absence of unique wild bird strains of C. jejuni in livestock suggests that the direction of infection is predominantly from livestock to wild birds. C. lari was detected mainly in wild birds sampled in an estuarine or coastal habitat. Fifteen C. lari isolates were analyzed by macrorestriction pulsed-field gel electrophoresis, which revealed genetically diverse populations of C. lari in Eurasian oystercatchers (Haematopus ostralegus) and clonal populations in magpies (Pica pica). PMID- 19286782 TI - Activation and transfer of the chromosomal phage resistance mechanism AbiV in Lactococcus lactis. AB - AbiV is a chromosomally encoded phage resistance mechanism that is silent in the wild-type phage-sensitive strain Lactococcus lactis subsp. cremoris MG1363. Spontaneous phage-resistant mutants of L. lactis MG1363 were analyzed by reverse transcriptase PCR and shown to express AbiV. This expression was related to a reorganization in the upstream region of abiV. Transfer of abiV between two lactococcal strains, most likely by conjugation, was also demonstrated. To our knowledge, this is the first report of natural transfer of a chromosomally encoded phage resistance mechanism. PMID- 19286783 TI - Negative fitness consequences and transmission dynamics of a heritable fungal symbiont of a parasitic wasp. AB - Heritable bacterial symbionts are widespread in insects and can have many important effects on host ecology and fitness. Fungal symbionts are also important in shaping their hosts' behavior, interactions, and evolution, but they have been largely overlooked. Experimental tests to determine the relevance of fungal symbionts to their insect hosts are currently extremely rare, and to our knowledge, there have been no such tests for strictly predacious insects. We investigated the fitness consequences for a parasitic wasp (Comperia merceti) of an inherited fungal symbiont in the Saccharomycotina (Ascomycota) that was long presumed to be a mutualist. In comparisons of wasp lines with and without this symbiont, we found no evidence of mutualism. Instead, there were significant fitness costs to the wasps in the presence of the yeast; infected wasps attacked fewer hosts and had longer development times. We also examined the relative competitive abilities of the larval progeny of infected and uninfected mothers, as well as horizontal transmission of the fungal symbiont among larval wasps that shared a single host cockroach egg case. We found no difference in larval competitive ability when larvae whose infection status differed shared a single host. We did find high rates of horizontal transmission of the fungus, and we suggest that this transmission is likely responsible for the maintenance of this infection in wasp populations. PMID- 19286784 TI - Antibody recognition force microscopy shows that outer membrane cytochromes OmcA and MtrC are expressed on the exterior surface of Shewanella oneidensis MR-1. AB - Antibody recognition force microscopy showed that OmcA and MtrC are expressed on the exterior surface of living Shewanella oneidensis MR-1 cells when Fe(III), including solid-phase hematite (Fe(2)O(3)), was the terminal electron acceptor. OmcA was localized to the interface between the cell and mineral. MtrC displayed a more uniform distribution across the cell surface. Both cytochromes were associated with an extracellular polymeric substance. PMID- 19286785 TI - Suppression of the bacterial spot pathogen Xanthomonas euvesicatoria on tomato leaves by an attenuated mutant of Xanthomonas perforans. AB - A bacteriocin-producing strain of the bacterial spot of tomato plant pathogen, Xanthomonas perforans, with attenuated pathogenicity was deployed for biocontrol of a bacteriocin-sensitive strain of the genetically closely related bacterial spot of tomato plant pathogen, X. euvesicatoria. The attenuated mutant (91 118DeltaopgHDeltabcnB) of X. perforans was tested in leaf tissue and shown to significantly inhibit internal populations of the wild-type X. euvesicatoria strain although significantly less than the wild-type 91-118 strain, whereas in a phyllosphere inhibition assay, the mutant strain reduced epiphytic populations comparably to 91-118. Thus, the attenuated mutant limited the sensitive bacterium more efficiently on the leaf surface than inside the leaf. In field experiments, weekly application of 91-118DeltaopgHDeltabcnB significantly reduced X. euvesicatoria populations compared to the growers' standard control (copper hydroxide and mancozeb applied weekly and acibenzolar-S-methyl applied every 2 weeks). The biological control agent, 91-118DeltaopgHDeltabcnB, applied every 2 weeks also significantly reduced X. euvesicatoria populations in one season but was not significantly different from the growers' standard control. Potentially, attenuated pathogenic strains could be deployed as biological control agents in order to improve disease control of foliar plant pathogens. PMID- 19286786 TI - prtH2, not prtH, is the ubiquitous cell wall proteinase gene in Lactobacillus helveticus. AB - Lactobacillus helveticus strains possess an efficient proteolytic system that releases peptides which are essential for lactobacillus growth in various fermented dairy products and also affect textural properties or biological activities. Cell envelope proteinases (CEPs) are bacterial enzymes that hydrolyze milk proteins. In the case of L. helveticus, two CEPs with low percentages of amino acid identity have been described, i.e., PrtH and PrtH2. However, the distribution of the genes that encode CEPs still remains unclear, rendering it difficult to further control the formation of particular peptides. This study evaluated the diversity of genes that encode CEPs in a collection of strains of L. helveticus isolated from various biotopes, both in terms of the presence or absence of these genes and in terms of nucleotide sequence, and studied their transcription in dairy matrices. After defining three sets of primers for both the prtH and prtH2 genes, we studied the distribution of the genes by using PCR and Southern blotting experiments. The prtH2 gene was ubiquitous in the 29 strains of L. helveticus studied, whereas only 18 of them also exhibited the prtH gene. Sequencing of a 350-bp internal fragment of these genes revealed the existence of intraspecific diversity. Finally, expression of these two CEP encoding genes was followed during the growth in dairy matrices of two strains, ITG LH77 and CNRZ32, which possess one and two CEP-encoding genes, respectively. Both genes were shown to be expressed by L. helveticus at each stage of growth in milk and at different stages of mini-Swiss-type cheese making and ripening. PMID- 19286787 TI - Candicidin biosynthesis gene cluster is widely distributed among Streptomyces spp. isolated from the sediments and the neuston layer of the Trondheim fjord, Norway. AB - A large number of Streptomyces bacteria with antifungal activity isolated from samples collected in the Trondheim fjord (Norway) were found to produce polyene compounds. Investigation of polyene-containing extracts revealed that most of the isolates produced the same compound, which had an atomic mass and UV spectrum corresponding to those of candicidin D. The morphological diversity of these isolates prompted us to speculate about the involvement of a mobile genetic element in dissemination of the candicidin biosynthesis gene cluster (can). Eight candicidin-producing isolates were analyzed by performing a 16S rRNA gene-based taxonomic analysis, pulsed-field gel electrophoresis, PCR, and Southern blot hybridization with can-specific probes. These analyses revealed that most of the isolates were related, although they were morphologically diverse, and that all of them contained can genes. The majority of the isolates studied contained large plasmids, and two can-specific probes hybridized to a 250-kb plasmid in one isolate. Incubation of the latter isolate at a high temperature resulted in loss of the can genes and candicidin production, while mating of the "cured" strain with a plasmid-containing donor restored candicidin production. The latter result suggested that the 250-kb plasmid contains the complete can gene cluster and could be responsible for conjugative transfer of this cluster to other streptomycetes. PMID- 19286788 TI - Characterization of a new plasmid-like prophage in a pandemic Vibrio parahaemolyticus O3:K6 strain. AB - Vibrio parahaemolyticus is a common food-borne pathogen that is normally associated with seafood. In 1996, a pandemic O3:K6 strain abruptly appeared and caused the first pandemic of this pathogen to spread throughout many Asian countries, America, Europe, and Africa. The role of temperate bacteriophages in the evolution of this pathogen is of great interest. In this work, a new temperate phage, VP882, from a pandemic O3:K6 strain of V. parahaemolyticus was purified and characterized after mitomycin C induction. VP882 was a Myoviridae bacteriophage with a polyhedral head and a long rigid tail with a sheath-like structure. It infected and lysed high proportions of V. parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae strains. The genome of phage VP882 was sequenced and was 38,197 bp long, and 71 putative open reading frames were identified, of which 27 were putative functional phage or bacterial genes. VP882 had a linear plasmid-like genome with a putative protelomerase gene and cohesive ends. The genome does not integrate into the host chromosome but was maintained as a plasmid in the lysogen. Analysis of the reaction sites of the protelomerases in different plasmid-like phages revealed that VP882 and PhiHAP-1 were highly similar, while N15, PhiKO2, and PY54 made up another closely related group. The presence of DNA adenine methylase and quorum-sensing transcriptional regulators in VP882 may play a specific role in this phage or regulate physiological or virulence-associated traits of the hosts. These genes may also be remnants from the bacterial chromosome following transduction. PMID- 19286789 TI - Dual role of the oligopeptide permease Opp3 during growth of Staphylococcus aureus in milk. AB - Staphylococcus aureus RN6390 presents a diauxic growth in milk, due to amino acid limitation. Inactivation of the oligopeptide permease Opp3 (dedicated to the nitrogen nutrition of the strain) not only affects the growth of the strain but also results in reduced expression levels of three major extracellular proteases. PMID- 19286790 TI - Mixed-species genomic microarray analysis of fecal samples reveals differential transcriptional responses of bifidobacteria in breast- and formula-fed infants. AB - Although their exact function remains enigmatic, bifidobacteria are among the first colonizers of the newborn infant gut and further develop into abundant communities, notably in response to diet. Therefore, the transcriptional responses of bifidobacteria in rapidly processed fecal samples from young infants that were fed either breast milk or a formula containing a mixture of galacto- and fructo-oligosaccharides were studied. The presence and diversity of the bifidobacterial fecal communities were determined using PCR-denaturing gradient gel electrophoresis and quantitative real-time PCR for specific species. Changes in the total number of bifidobacteria as well as in species diversity were observed, indicating the metabolic activities of the bifidobacteria within the infant gut. In addition, total RNAs isolated from infant feces were labeled and hybridized to a bifidobacterium-specific microarray comprising approximately 6,000 clones of the major bifidobacterial species of the human gut. Approximately 270 clones that showed the most prominent hybridization with the samples were sequenced. Fewer than 10% of the hybridizing clones contained rRNA genes, whereas the vast majority of the inserts showed matches with protein-encoding genes predicted to originate from bifidobacteria. Although a wide range of functional groups was covered by the obtained sequences, the largest fraction (14%) of the transcribed genes assigned to a functional category were predicted to be involved in carbohydrate metabolism, while some were also implicated in exopolysaccharide production or folate production. A total of three of the above-described protein encoding genes were selected for quantitative PCR and sequence analyses, which confirmed the expression of the corresponding genes and the expected nucleotide sequences. In conclusion, the results of this study show the feasibility of obtaining insight into the transcriptional responses of intestinal bifidobacteria by analyzing fecal RNA and highlight the in vivo expression of bifidobacterial genes implicated in host-related functions. PMID- 19286791 TI - High rate of N2 fixation by East Siberian cryophilic soil bacteria as determined by measuring acetylene reduction in nitrogen-poor medium solidified with gellan gum. AB - For evaluating N(2) fixation of diazotrophic bacteria, nitrogen-poor liquid media supplemented with at least 0.5% sugar and 0.2% agar are widely used for acetylene reduction assays. In such a soft gel medium, however, many N(2)-fixing soil bacteria generally show only trace acetylene reduction activity. Here, we report that use of a N(2) fixation medium solidified with gellan gum instead of agar promoted growth of some gellan-preferring soil bacteria. In a soft gel medium solidified with 0.3% gellan gum under appropriate culture conditions, bacterial microbiota from boreal forest bed soils and some free-living N(2)-fixing soil bacteria isolated from the microbiota exhibited 10- to 200-fold-higher acetylene reduction than those cultured in 0.2% agar medium. To determine the N(2) fixation activating mechanism of gellan gum medium, qualitative differences in the colony forming bacterial components from tested soil microbiota were investigated in plate cultures solidified with either agar or gellan gum for use with modified Winogradsky's medium. On 1.5% agar plates, apparently cryophilic bacterial microbiota showed strictly distinguishable microbiota according to the depth of soil in samples from an eastern Siberian Taiga forest bed. Some pure cultures of proteobacteria, such as Pseudomonas fluorescens and Burkholderia xenovorans, showed remarkable acetylene reduction. On plates solidified with 1.0% gellan gum, some soil bacteria, including Luteibacter sp., Janthinobacterium sp., Paenibacillus sp., and Arthrobacter sp., uniquely grew that had not grown in the presence of the same inoculants on agar plates. In contrast, Pseudomonas spp. and Burkholderia spp. were apparent only as minor colonies on the gellan gum plates. Moreover, only gellan gum plates allowed some bacteria, particularly those isolated from the shallow organic soil layer, to actively swarm. In consequence, gellan gum is a useful gel matrix to bring out growth potential capabilities of many soil diazotrophs and their consortia in communities of soil bacteria. PMID- 19286792 TI - Comparison of planktonic and biofilm cultures of Pseudomonas fluorescens DSM 8341 cells grown on fluoroacetate. AB - Comparisons between the physiological properties of Pseudomonas fluorescens biofilm cells grown in a tubular biofilm reactor and planktonic cells grown in a chemostat were performed. Fluoroacetate was the sole carbon source for all experiments. The performance of cells was assessed using cell cycle kinetics and by determining specific fluoroacetate utilization rates. Cell cycle kinetics were studied by flow cytometry in conjunction with the fluorescent stain propidium iodide. Determination of the DNA content of planktonic and biofilm cultures showed little difference between the two modes of growth. Cultures with comparable specific glycolate utilization rates had similar percentages of cells in the B phase of the cell cycle, indicating similar growth rates. Specific fluoroacetate utilization rates showed the performance of planktonic cells to be superior to that of biofilm cells, with more fluoroacetate utilized per cell at similar specific fluoroacetate loading rates. A consequence of this decreased biofilm performance was the accumulation of glycolate in the effluent of biofilm cultures. This accumulation of glycolate was not observed in the effluent of planktonic cultures. Spatial stratification of oxygen within the biofilm was identified as a possible explanation for the overflow metabolism of glycolate and the decreased performance of the biofilm cells. PMID- 19286793 TI - Global distribution and evolution of a toxinogenic Burkholderia-Rhizopus symbiosis. AB - Toxinogenic endobacteria were isolated from a collection of Rhizopus spp. representing highly diverse geographic origins and ecological niches. All endosymbionts belonged to the Burkholderia rhizoxinica complex according to matrix-assisted laser desorption ionization-time of flight biotyping and multilocus sequence typing, suggesting a common ancestor. Comparison of host and symbiont phylogenies provides insights into possible cospeciation and horizontal transmission events. PMID- 19286794 TI - Osmoadaptation among Vibrio species and unique genomic features and physiological responses of Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus is a moderately halophilic bacterium found in estuarine and marine coastal ecosystems worldwide. Although the ability of V. parahaemolyticus to grow and proliferate in fluctuating saline environments is well known, the underlying molecular mechanisms of osmoadaptation are unknown. We performed an in silico analysis of V. parahaemolyticus strain RIMD2210633 for genes homologous to osmotic stress response genes in other bacteria. We uncovered two putative compatible solute synthesis systems (encoded by ectABC and betABI) and six putative compatible solute transporters (encoded by four bcct loci and two proVWX loci). An ectoine synthesis system clustered with a betaine/carnitine/choline transporter and a ProU transporter (encoded by homologues of proVWX from Escherichia coli), and a betaine synthesis system clustered with a ProU transporter (encoded by homologues of proVXW from Pseudomonas syringae). This is at least double the number present in V. cholerae, V. fischeri, or V. vulnificus. Six additional Vibrio species contain both ectABC and betABI, i.e., V. alginolyticus 12G01, V. angustum, V. harveyi BAA-1116, V. splendidus LGP32, Vibrio sp. strain MED222, and Vibrio sp. strain Ex25. V. harveyi HY01 and V. splendidus 12B01 only encoded the betaine system. In addition, V. alginolyticus had a compendium of systems identical to that found in V. parahaemolyticus. Comparative physiological analysis of RIMD2210633 with V. vulnificus YJ016, V. cholerae N16961, and V. fischeri ES114 grown at different salinities and temperatures demonstrated that V. parahaemolyticus had a growth advantage under all of the conditions examined. We demonstrate, by one dimensional nuclear magnetic resonance analysis, that V. parahaemolyticus is capable of de novo synthesis of ectoine at high salinity whereas a Delta ectB knockout strain is not. We constructed a single-knockout mutation in proU1, but no growth defect was noted, indicating transporter system redundancy. We complemented E. coli MKH13, a compatible solute transporter-negative strain, with bcct2 and demonstrated uptake of betaine at high salt concentrations. PMID- 19286795 TI - Migratory response of soil bacteria to Lyophyllum sp. strain Karsten in soil microcosms. AB - In this study, the selection of bacteria on the basis of their migration via fungal hyphae in soil was investigated in microcosm experiments containing Lyophyllum sp. strain Karsten (DSM2979). One week following inoculation with a bacterial community obtained from soil, selection of a few specific bacterial types was noticed at 30 mm in the growth direction of Lyophyllum sp. strain Karsten in sterile soil. Cultivation-based analyses showed that the migration proficient types encompassed 10 bacterial groups, as evidenced by (GTG)(5) genomic fingerprinting as well as 16S rRNA gene sequencing. These were (>97% similarity) Burkholderia terrae BS001, Burkholderia sordidicola BS026, Burkholderia sediminicola BS010, and Burkholderia phenazinium BS028; Dyella japonica BS013, BS018, and BS021; "Sphingoterrabacterium pocheensis" BS024; Sphingobacterium daejeonense BS025; and Ralstonia basilensis BS017. Migration as single species was subsequently found for B. terrae BS001, D. japonica BS018 and BS021, and R. basilensis BS017. Typically, migration occurred only when these organisms were introduced at the fungal growth front and only in the direction of hyphal growth. Migration proficiency showed a one-sided correlation with the presence of the hrcR gene, used as a marker for the type III secretion system (TTSS), as all single-strain migrators were equipped with this system and most non-single-strain migrators were not. The presence of the TTSS stood in contrast to the low prevalence of TTSSs within the bacterial community used as an inoculum (<3%). Microscopic examination of B. terrae BS001 in contact with Lyophyllum sp. strain Karsten hyphae revealed the development of a biofilm surrounding the hyphae. Migration-proficient bacteria interacting with Lyophyllum sp. strain Karsten may show complex behavior (biofilm formation) at the fungal tip, leading to their translocation and growth in novel microhabitats in soil. PMID- 19286796 TI - Proliferation of antibiotic-producing bacteria and concomitant antibiotic production as the basis for the antibiotic activity of Jordan's red soils. AB - Anecdotes, both historical and recent, recount the curing of skin infections, including diaper rash, by using red soils from the Hashemite Kingdom of Jordan. Following inoculation of red soils isolated from geographically separate areas of Jordan, Micrococcus luteus and Staphylococcus aureus were rapidly killed. Over the 3-week incubation period, the number of specific types of antibiotic producing bacteria increased, and high antimicrobial activity (MIC, approximately 10 microg/ml) was observed in methanol extracts of the inoculated red soils. Antibiotic-producing microorganisms whose numbers increased during incubation included actinomycetes, Lysobacter spp., and Bacillus spp. The actinomycetes produced actinomycin C(2) and actinomycin C(3). No myxobacteria or lytic bacteriophages with activity against either M. luteus or S. aureus were detected in either soil before or after inoculation and incubation. Although protozoa and amoebae were detected in the soils, the numbers were low and did not increase over the incubation period. These results suggest that the antibiotic activity of Jordan's red soils is due to the proliferation of antibiotic-producing bacteria. PMID- 19286797 TI - Glycerol utilization gene cluster in Streptomyces clavuligerus. AB - The Streptomyces clavuligerus ATCC 27064 glycerol cluster gylR-glpF1K1D1 is induced by glycerol but is not affected by glucose. S. clavuligerus growth and clavulanic acid production are stimulated by glycerol, but this does not occur in a glpK1-deleted mutant. Amplification of glpK1D1 results in transformants yielding larger amounts of clavulanic acid in the wild-type strain and in overproducer S. clavuligerus Gap15-7-30 or S. clavuligerus Delta relA strains. PMID- 19286798 TI - Competence for genetic transformation in Streptococcus pneumoniae: termination of activity of the alternative sigma factor ComX is independent of proteolysis of ComX and ComW. AB - Competence for genetic transformation in Streptococcus pneumoniae is a transient physiological state whose development is coordinated by a peptide pheromone (CSP) and its receptor, which activates transcription of two downstream genes, comX and comW, and 15 other "early" genes. ComX, a transient alternative sigma factor, drives transcription of "late" genes, many of which are essential for transformation. In vivo, ComW both stabilizes ComX against proteolysis by the ClpE-ClpP protease and stimulates its activity. Interestingly, stabilization of ComX by deletion of the gene encoding the ClpP protease did not extend the period of competence. We considered the hypothesis that the rapid decay of competence arises from a rapid loss of ComW and thus of its ComX stimulating activity, so that ComX might persist but lose its transcriptional activity. Western analysis revealed that ComW is indeed a transient protein, which is also stabilized by deletion of the gene encoding the ClpP protease. However, stabilizing both ComX and ComW did not prolong either ComX activity or the period of transformation, indicating that termination of the transcriptional activity of ComX is not dependent on proteolysis of ComW. PMID- 19286799 TI - Identification and characterization of inorganic pyrophosphatase and PAP phosphatase from Thermococcus onnurineus NA1. AB - Two hypothetical genes were functionally verified to be a pyrophosphatase and a PAP phosphatase in Thermococcus onnurineus NA1. This is the first report of the pyrophosphatases and the PAP phosphatases being organized in the gene clusters of the sulfate activation system only in T. onnurineus NA1 and "Pyrococcus abyssi." PMID- 19286800 TI - Whole-genome tiling array analysis of Mycobacterium leprae RNA reveals high expression of pseudogenes and noncoding regions. AB - Whole-genome sequence analysis of Mycobacterium leprae has revealed a limited number of protein-coding genes, with half of the genome composed of pseudogenes and noncoding regions. We previously showed that some M. leprae pseudogenes are transcribed at high levels and that their expression levels change following infection. In order to clarify the RNA expression profile of the M. leprae genome, a tiling array in which overlapping 60-mer probes cover the entire 3.3 Mbp genome was designed. The array was hybridized with M. leprae RNA from the SHR/NCrj-rnu nude rat, and the results were compared to results from an open reading frame array and confirmed by reverse transcription-PCR. RNA expression was detected from genes, pseudogenes, and noncoding regions. The signal intensities obtained from noncoding regions were higher than those from pseudogenes. Expressed noncoding regions include the M. leprae unique repetitive sequence RLEP and other sequences without any homology to known functional noncoding RNAs. Although the biological functions of RNA transcribed from M. leprae pseudogenes and noncoding regions are not known, RNA expression analysis will provide insights into the bacteriological significance of the species. In addition, our study suggests that M. leprae will be a useful model organism for the study of the molecular mechanism underlying the creation of pseudogenes and the role of microRNAs derived from noncoding regions. PMID- 19286801 TI - Structural requirements of the Pseudomonas quinolone signal for membrane vesicle stimulation. AB - Pseudomonas aeruginosa produces the quorum signal 2-heptyl-3-hydroxy-4-quinolone (Pseudomonas quinolone signal), which is important for stimulating outer membrane vesicle (MV) formation. Here we describe the importance of the 3-hydroxyl and 2 alkyl chain for MV production and the length of the 2-alkyl chain for association with MVs. PMID- 19286802 TI - Mechanism for sortase localization and the role of sortase localization in efficient pilus assembly in Enterococcus faecalis. AB - Pathogenic streptococci and enterococci primarily rely on the conserved secretory (Sec) pathway for the translocation and secretion of virulence factors out of the cell. Since many secreted virulence factors in gram-positive organisms are subsequently attached to the bacterial cell surface via sortase enzymes, we sought to investigate the spatial relationship between secretion and cell wall attachment in Enterococcus faecalis. We discovered that sortase A (SrtA) and sortase C (SrtC) are colocalized with SecA at single foci in the enterococcus. The SrtA-processed substrate aggregation substance accumulated in single foci when SrtA was deleted, implying a single site of secretion for these proteins. Furthermore, in the absence of the pilus-polymerizing SrtC, pilin subunits also accumulate in single foci. Proteins that localized to single foci in E. faecalis were found to share a positively charged domain flanking a transmembrane helix. Mutation or deletion of this domain in SrtC abolished both its retention at single foci and its function in efficient pilus assembly. We conclude that this positively charged domain can act as a localization retention signal for the focal compartmentalization of membrane proteins. PMID- 19286803 TI - Regulation of superoxide dismutase (sod) genes by SarA in Staphylococcus aureus. AB - The scavenging of reactive oxygen species (ROS) within cells is regulated by several interacting factors, including transcriptional regulators. Involvement of sarA family genes in the regulation of proteins involved in the scavenging of ROS is largely unknown. In this report, we show that under aerobic conditions, the levels of sodM and sodA transcription, in particular the sodM transcript, are markedly enhanced in the sarA mutant among the tested sarA family mutants. Increased levels of sod expression returned to near the parental level in a single-copy sarA complemented strain. Under microaerophilc conditions, transcription of both sodM and sodA was considerably enhanced in the sarA mutant compared to the wild-type strain. Various genotypic, phenotypic, and DNA binding studies confirmed the involvement of SarA in the regulation of sod transcripts in different strains of Staphylococcus aureus. The sodA mutant was sensitive to an oxidative stress-inducing agent, methyl viologen, but the sarA sodA double mutant was more resistant to the same stressor than the single sodA mutant. These results suggest that overexpression of SodM, which occurs in the sarA background, can rescue the methyl viologen-sensitive phenotype observed in the absence of the sodA gene. Analysis with various oxidative stress-inducing agents indicates that SarA may play a greater role in modulating oxidative stress resistance in S. aureus. This is the first report that demonstrates the direct involvement of a regulatory protein (SarA) in control of sod expression in S. aureus. PMID- 19286804 TI - Nitrite transport activity of the ABC-type cyanate transporter of the cyanobacterium Synechococcus elongatus. AB - In addition to the ATP-binding cassette (ABC)-type nitrate/nitrite-bispecific transporter, which has a high affinity for both substrates (K(m), approximately 1 microM), Synechococcus elongatus has an active nitrite transport system with an apparent K(m) (NO(2)(-)) value of 20 microM. We found that this activity depends on the cynABD genes, which encode a putative cyanate (NCO(-)) ABC-type transporter. Accordingly, nitrite transport by CynABD was competitively inhibited by NCO(-) with a K(i) value of 0.025 microM. The transporter was induced under conditions of nitrogen deficiency, and the induced cells showed a V(max) value of 11 to 13 micromol/mg of chlorophyll per h for cyanate or nitrite, which could supply approximately 30% of the amount of nitrogen required for optimum growth. Its relative specificity for the substrates and regulation at transcriptional and posttranslational levels suggested that the physiological role of the bispecific cyanate/nitrite transporter in S. elongatus is to allow nitrogen-deficient cells to assimilate low concentrations of cyanate in the medium. Its contribution to nitrite assimilation was significant in a mutant lacking the ABC-type nitrate/nitrite transporter, suggesting a possible role for CynABD in nitrite assimilation by cyanobacterial species that lack another high-affinity mechanism(s) for nitrite transport. PMID- 19286805 TI - Oxidation of cysteine 645 of cobalamin-independent methionine synthase causes a methionine limitation in Escherichia coli. AB - Cobalamin-independent methionine synthase (MetE) catalyzes the final step in Escherichia coli methionine biosynthesis but is inactivated under oxidative conditions, triggering a methionine deficiency. This study demonstrates that the mutation of MetE cysteine 645 to alanine completely eliminates the methionine auxotrophy imposed by diamide treatment, suggesting that modulation of MetE activity via cysteine 645 oxidation has significant physiological consequences for oxidatively stressed cells. PMID- 19286806 TI - Cytoskeletal asymmetrical dumbbell structure of a gliding mycoplasma, Mycoplasma gallisepticum, revealed by negative-staining electron microscopy. AB - Several mycoplasma species feature a membrane protrusion at a cell pole, and unknown mechanisms provide gliding motility in the direction of the pole defined by the protrusion. Mycoplasma gallisepticum, an avian pathogen, is known to form a membrane protrusion composed of bleb and infrableb and to glide. Here, we analyzed the gliding motility of M. gallisepticum cells in detail. They glided in the direction of the bleb at an average speed of 0.4 microm/s and remained attached around the bleb to a glass surface, suggesting that the gliding mechanism is similar to that of a related species, Mycoplasma pneumoniae. Next, to elucidate the cytoskeletal structure of M. gallisepticum, we stripped the envelopes by treatment with Triton X-100 under various conditions and observed the remaining structure by negative-staining transmission electron microscopy. A unique cytoskeletal structure, about 300 nm long and 100 nm wide, was found in the bleb and infrableb. The structure, resembling an asymmetrical dumbbell, is composed of five major parts from the distal end: a cap, a small oval, a rod, a large oval, and a bowl. Sonication likely divided the asymmetrical dumbbell into a core and other structures. The cytoskeletal structures of M. gallisepticum were compared with those of M. pneumoniae in detail, and the possible protein components of these structures were considered. PMID- 19286807 TI - Crystal structure of a chimeric receptor binding protein constructed from two lactococcal phages. AB - Lactococcus lactis, a gram-positive bacterium widely used by the dairy industry to manufacture cheeses, is subject to infection by a diverse population of virulent phages. We have previously determined the structures of three receptor binding proteins (RBPs) from lactococcal phages TP901-1, p2, and bIL170, each of them having a distinct host range. Virulent phages p2 and bIL170 are classified within the 936 group, while the temperate phage TP901-1 is a member of the genetically distinct P335 polythetic group. These RBPs comprise three domains: the N-terminal domain, binding to the virion particle; a beta-helical linker domain; and the C-terminal domain, bearing the receptor binding site used for host recognition. Here, we have designed, expressed, and determined the structure of an RBP chimera in which the N-terminal and linker RBP domains of phage TP901-1 (P335) are fused to the C-terminal RBP domain of phage p2 (936). This chimera exhibits a stable structure that closely resembles the parental structures, while a slight displacement of the linker made RBP domain adaptation efficient. The receptor binding site is structurally indistinguishable from that of native p2 RBP and binds glycerol with excellent affinity. PMID- 19286808 TI - Identification and characterization of a novel multidrug resistance operon, mdtRP (yusOP), of Bacillus subtilis. AB - Using comparative genome sequencing analysis, we identified a novel mutation in Bacillus subtilis that confers a low level of resistance to fusidic acid. This mutation was located in the mdtR (formerly yusO) gene, which encodes a MarR-type transcriptional regulator, and conferred a low level of resistance to several antibiotics, including novobiocin, streptomycin, and actinomycin D. Transformation experiments showed that this mdtR mutation was responsible for multidrug resistance. Northern blot analysis revealed that the downstream gene mdtP (formerly yusP), which encodes a multidrug efflux transporter, is cotranscribed with mdtR as an operon. Disruption of the mdtP gene completely abolished the multidrug resistance phenotype observed in the mdtR mutant. DNase I footprinting and primer extension analyses demonstrated that the MdtR protein binds directly to the mdtRP promoter, thus leading to repression of its transcription. Moreover, gel mobility shift analysis indicated that an Arg83 --> Lys or Ala67 --> Thr substitution in MdtR significantly reduces binding affinity to DNA, resulting in derepression of mdtRP transcription. Low concentrations of fusidic acid induced the expression of mdtP, although the level of mdtP expression was much lower than that in the mdtR disruptant. These findings indicate that the MdtR protein is a repressor of the mdtRP operon and that the MdtP protein functions as a multidrug efflux transporter in B. subtilis. PMID- 19286809 TI - A novel nitroreductase of Staphylococcus aureus with S-nitrosoglutathione reductase activity. AB - In this report we show that inactivation of the putative nitroreductase SA0UHSC_00833 (ntrA) increases the sensitivity of Staphylococcus aureus to S nitrosoglutathione (GSNO) and augments its resistance to nitrofurans. S. aureus NtrA is a bifunctional enzyme that exhibits nitroreductase and GSNO reductase activity. A phylogenetic analysis suggests that NtrA is a member of a novel family of nitroreductases that seems to play a dual role in vivo, promoting nitrofuran activation and protecting the cell against transnitrosylation. PMID- 19286810 TI - Large intergenic cruciform-like supermotifs in the Lactobacillus plantarum genome. AB - Twenty-four Lactobacillus plantarum supermotifs (LPSMs) with lengths from approximately 800 to 1,000 nucleotides were identified in the L. plantarum genome. LPSMs were conserved in other L. plantarum strains but not in other species. Secondary structure analysis predicted that LPSMs may fold into cruciform-like structures. Preliminary experiments indicate that the LPSMs are transcribed. PMID- 19286811 TI - Thioredoxins in redox maintenance and survival during oxidative stress of Bacteroides fragilis. AB - The anaerobe Bacteroides fragilis is a gram-negative, opportunistic pathogen that is highly aerotolerant and can persist in aerobic environments for extended periods. In this study, the six B. fragilis thioredoxins (Trxs) were investigated to determine their role during oxidative stress. Phylogenetic analyses of Trx protein sequences indicated that four of the six Trxs (TrxA, TrxC, TrxD, and TrxF) belong to the M-type Trx class but were associated with two different M type lineages. TrxE and TrxG were most closely associated to Y-type Trxs found primarily in cyanobacteria. Single and multiple trx gene deletions were generated to determine functional differences between the Trxs. The trxA gene was essential, but no anaerobic growth defects were observed for any other single trx deletion or for the DeltatrxC DeltatrxD::cfxA DeltatrxE DeltatrxF DeltatrxG quintuple mutant. Regulation of the trx genes was linked to the oxidative stress response, and all were induced by aerobic conditions. The DeltatrxC DeltatrxE DeltatrxF DeltatrxG and the DeltatrxC DeltatrxD::cfxA DeltatrxE DeltatrxF DeltatrxG multiple deletion strains were impaired during growth in oxidized media, but single trx gene mutants did not have a phenotype in this assay. TrxD was protective during exposure to the thiol oxidant diamide, and expression of trxD was induced by diamide. Diamide-induced expression of trxC, trxE, and trxF increased significantly in a trxD mutant strain, suggesting that there is some capacity for compensation in this complex Trx system. These data provide insight into the role of individual Trxs in the B. fragilis oxidative stress response. PMID- 19286812 TI - Genetic parameters and trends for lamb survival and birth weight in a Merino flock divergently selected for multiple rearing ability. AB - Data of 5,390 Merino lambs born from 1986 to 2007 (approximately 6.9 generations) were used to derive genetic parameters and trends for age-specific and overall lamb survival on the underlying scale, as well as for lamb birth weight, using Gibbs sampling. The majority of lambs were descended from lines that were divergently selected for the ability of ewes to rear multiples. The line selected in the upward direction was denoted as the high line (H line), whereas the line selected in the downward direction was the low line (L line). Analyses included the covariance between direct and maternal genetic effects, except where it was not estimable owing to small direct additive variance components, a high incidence of lambs surviving, or both. Direct heritability estimates were 0.02 for lamb survival at birth, 0.12 for lamb survival from birth to tail docking, 0.39 for lamb survival from docking to weaning, 0.28 for overall lamb survival, and 0.17 for birth weight. Corresponding estimates for the maternal genetic effect were 0.26, 0.14, 0.16, 0.14, and 0.29. Dam permanent environmental variance ratios were, respectively, 0.14, 0.09, 0.05, 0.07, and 0.07. Estimates of the direct-maternal genetic correlation were -0.60 for lamb survival from docking to weaning, -0.61 for overall lamb survival, and -0.15 to -0.23 for birth weight. Genetic, maternal genetic, and dam permanent environmental correlations between lamb birth weight and overall or age-specific lamb survival were not different from zero. Expressed relative to overall means, annual direct genetic change in the H line amounted to -0.01% per annum for lamb survival at birth, 0.52% per annum for lamb survival from birth to docking, and 1.3% per annum for lamb survival from docking to weaning. Corresponding trends in the L line were, respectively, -0.01, -0.42, and 0.042% per annum. Maternal genetic trends amounted to, respectively, 0.23, -0.11, and -0.24% per annum in the H line and 0.78, -0.50, and -0.16% per annum in the L line. It was concluded that sustained genetic progress in lamb survival is feasible if directed selection is applied to a correlated trait such as the ability of ewes to rear multiples. PMID- 19286813 TI - Bioaccumulation of ergovaline in bovine lateral saphenous veins in vitro. AB - Ergot alkaloids have been associated with vasoconstriction in grazing livestock affected by the fescue toxicosis syndrome. Previous in vitro investigations studying how ergot alkaloids caused vasoconstriction have shown that ergovaline has a distinct receptor affinity and sustained contractile response. A similar contractile response has not been noted for lysergic acid. The objectives of this study were to determine if repetitive in vitro exposure of bovine lateral saphenous vein to lysergic acid or ergovaline would result in an increasing contractile response and if a measurable bioaccumulation of the alkaloids in the vascular tissue occurs over time. Segments of vein were surgically biopsied from healthy, Angus x Brangus cross-bred, fescue-naive yearling heifers (n = 16) or collected from healthy mixed breed and sex cattle immediately after slaughter (n = 12) at a local abattoir. Veins were trimmed of excess fat and connective tissue, sliced into cross-sections, and suspended in a myograph chamber containing 5 mL of oxygenated Krebs-Henseleit buffer (95% O(2)/5% CO(2); pH = 7.4; 37 degrees C). Contractile responses to repetitive additions of ergovaline (1 x 10(-9) and 1 x 10(-7) M) and lysergic acid (1 x 10(-5) and 1 x 10(-4) M) were evaluated using the biopsied veins. For the bioaccumulation experiments, veins collected at the abattoir underwent repetitive additions of 1 x 10(-7) M ergovaline and 1 x 10(-5) M lysergic acid and the segments were removed after every 2 additions and media rinses for alkaloid quantification via HPLC/mass spectrometry. Contractile data were normalized as a percentage of contractile response induced by a reference dose of norepinephrine (1 x 10(-4) M). Repetitive additions of 1 x 10(-9) M ergovaline and 1 x 10(-5) and 1 x 10(-4) M lysergic acid resulted in contractile response with a negative slope (P < 0.02). In contrast, repetitive addition of 1 x 10(-7) M ergovaline resulted in a contractile response that increased with each addition (P < 0.01). Lysergic acid and ergovaline were detected at all 4 exposure levels (2x to 8x), but only the 1 x 10(-7) M ergovaline treatment resulted in increased tissue content as the number of exposures increased (P < 0.05). These data indicate that ergovaline, but not lysergic acid, bioaccumulates with repetitive exposure in vitro. These results suggest that ergovaline may have a greater potential for inducing toxicosis in grazing animals than lysergic acid because of its potential to bioaccumulate at the cellular site of action. PMID- 19286814 TI - Effects of dietary vitamin supplementation and semen collection frequency on reproductive performance and semen quality in boars. AB - The present study was undertaken to assess the relevance of increasing the daily provision of dietary vitamins on vitamin metabolic status and semen characteristics of boars under controlled and commercial conditions as well as to evaluate the efficiency of this vitamin supplement to allow boars to cope with intensive semen collection frequency. In the first experiment, 39 boars were allocated to 2 dietary treatments, a basal diet (control) and the basal diet supplemented with extra fat- and water-soluble vitamins (Vit). Within each treatment, boars were submitted to 2 regimens of semen collection frequency: 3 times per 2 wk (3/2) and 3 times per week (3/1) over a 12-wk period. Afterwards, all boars were intensively collected (daily) for 2 wk. A resting period of 4 wk followed, and all boars were collected 2 times per week. Thereafter, collection frequencies were reversed, and the same procedure was followed until the end of the intensive collection period. A second experiment was conducted in commercial conditions at a commercial stud, and 252 boars were randomly allocated to the control and Vit dietary treatments. All boars were collected 2 times per week over a 6-mo period. Classical measurements of ejaculate and sperm quality were assessed, and blood samples were collected throughout both experiments to quantify vitamin concentrations. In the first experiment, vitamin concentrations in blood and seminal plasma increased in Vit boars (P < 0.05); however, vitamin concentrations were not affected by collection frequency (P > 0.14). The Vit supplement did not affect sperm production or sperm quality (P > 0.28), although semen volume increased during the 12-wk periods for Vit boars (P < 0.05). The 3/1 boars produced fewer doses per ejaculate than 3/2 boars (P < 0.01); however, the cumulative sperm production for the 12-wk periods increased by 19% in 3/1 boars compared with 3/2 boars. In the second experiment, blood plasma concentrations of vitamin B(9) were greater (P < 0.01) in Vit than control boars. The vitamin supplement did not increase sperm production of boars (P > 0.61). In conclusion, dietary supplements of fat- and water-soluble vitamins increase the amount of vitamins available for the animal, and the collection frequencies had no effect on vitamin status. Moreover, in spite of an effect on the ejaculate volume, the dietary supplement of extra vitamins had no effect on sperm production or quality. PMID- 19286815 TI - Effects of on-arrival versus delayed clostridial or modified live respiratory vaccinations on health, performance, bovine viral diarrhea virus type I titers, and stress and immune measures of newly received beef calves. AB - Stress, commonly associated with weaning, marketing, and shipment of feeder cattle, can compromise immune function, and vaccine administration during immunosuppression may reduce vaccine efficacy and calf growth. Four treatments were compared in a 2 x 2 factorial arrangement to evaluate the effect of on arrival (d 0) vs. delayed (d 14) administration of clostridial (CLOS) and respiratory (RESP) vaccines on health, performance, bovine viral diarrhea virus (BVDV) antibody titers, and physiological immune measurements of high-risk, newly received calves. Crossbred bull and steer calves (n = 263) were weighed (239 +/- 1.2 kg), stratified by sex, and randomly assigned to vaccination treatment: 1) arrival CLOS, arrival RESP (ACAR); 2) arrival CLOS, delayed RESP (ACDR); 3) delayed CLOS, arrival RESP (DCAR); and 4) delayed CLOS, delayed RESP (DCDR). Body weight and blood samples were collected on d 0, 14, 28, 42, and 56. Average daily gain did not differ (P > or = 0.34), averaging 0.98, 0.93, 0.95, and 0.91 kg/d for ACAR, ACDR, DCAR, and DCDR, respectively, for the entire 56-d trial. Vaccination timing did not affect morbidity (P > or = 0.23); however, there tended to be a CLOS timing effect (P = 0.07) and RESP timing effect (P = 0.09) on days to initial bovine respiratory disease (BRD) treatment. Average days to initial BRD treatment were less for ACAR (6 +/- 0.8 d) compared with DCDR (8 +/- 0.8 d; P = 0.01). Greater white blood cell counts were observed for DCDR than ACDR (P = 0.01), with ACAR and DCAR being intermediate. Serum cortisol concentrations were greater on d 0 than d 14 (P < 0.01) or d 28 (P = 0.01) but no treatment x day interaction (P = 0.21) was observed. Timing of RESP administration affected (P = 0.001) serum BVDV type I titers, with greater (P < 0.01) levels in calves receiving RESP vaccine on arrival. Delaying CLOS or RESP vaccination did not affect BW gain or morbidity in high risk, newly received stocker calves. Calves administered RESP vaccine on d 0 developed antibody titers to BVDV type I earlier than delayed RESP treatments. Total white blood cell count was greatest when RESP and CLOS vaccination were delayed (DCDR). PMID- 19286816 TI - The value of reproductive tract scoring as a predictor of fertility and production outcomes in beef heifers. AB - In this study, 272 beef heifers were studied from just before their first breeding season (October 15, 2003), through their second breeding season, and until just after they had weaned their first calves in March, 2005. This study was performed concurrently with another study testing the economic effects of an estrous synchronization protocol using PG. Reproductive tract scoring (RTS) by rectal palpation was performed on the group of heifers 1 d before the onset of their first breeding season. The effect of RTS on several fertility and production outcomes was tested, and the association of RTS with the outcomes was compared with that of other input variables such as BW, age, BCS, and Kleiber ratio using multiple or univariable linear, logistic, or Cox regression. Area under the curve for receiver operating characteristic analysis was used to compare the ability of different input variables to predict pregnancy outcome. After adjustment for BW and age, RTS was positively associated with pregnancy rate to the 50-d AI season (P < 0.01), calf weaning weight (r = 0.22, P < 0.01), and pregnancy rate to the subsequent breeding season (P < 0.01), and negatively associated with days to calving (r = 0.28, P < 0.01). Reproductive tract scoring was a better predictor of fertility than was Kleiber ratio and similar in its prediction of calf weaning weight. It was concluded from this study that RTS is a predictor of heifer fertility, compares well with other traits used as a predictor of production outcomes, and is likely to be a good predictor of lifetime production of the cow. PMID- 19286817 TI - The environmental impact of dairy production: 1944 compared with 2007. AB - A common perception is that pasture-based, low-input dairy systems characteristic of the 1940s were more conducive to environmental stewardship than modern milk production systems. The objective of this study was to compare the environmental impact of modern (2007) US dairy production with historical production practices as exemplified by the US dairy system in 1944. A deterministic model based on the metabolism and nutrient requirements of the dairy herd was used to estimate resource inputs and waste outputs per billion kg of milk. Both the modern and historical production systems were modeled using characteristic management practices, herd population dynamics, and production data from US dairy farms. Modern dairy practices require considerably fewer resources than dairying in 1944 with 21% of animals, 23% of feedstuffs, 35% of the water, and only 10% of the land required to produce the same 1 billion kg of milk. Waste outputs were similarly reduced, with modern dairy systems producing 24% of the manure, 43% of CH(4), and 56% of N(2)O per billion kg of milk compared with equivalent milk from historical dairying. The carbon footprint per billion kilograms of milk produced in 2007 was 37% of equivalent milk production in 1944. To fulfill the increasing requirements of the US population for dairy products, it is essential to adopt management practices and technologies that improve productive efficiency, allowing milk production to be increased while reducing resource use and mitigating environmental impact. PMID- 19286818 TI - Impact of water troughs on cattle use of riparian zones in the Georgia Piedmont in the United States. AB - Cattle use of riparian areas may lead to stream water contamination with nutrients, pathogens, and sediments. Providing alternative water away from the stream may reduce the amount of time cattle spend near streams and therefore reduce contamination. We conducted this study to 1) evaluate the effect of providing water troughs outside of the riparian zones on the amount of time cattle spend in riparian zones, and 2) evaluate if environmental factors such as temperature and humidity affect the impact of water trough availability on the amount of time cattle spend within riparian and nonriparian locations. Global positioning system (GPS) collars were used to document cow locations every 5 min in 2 mixed tall fescue/common bermuda-grass pastures of the Georgia Piedmont in the United States. We found that when the temperature and humidity index (THI) ranged between 62 and 72, providing cattle with water troughs outside of riparian zones tended to decrease time cattle spent in riparian zones by 63% (52 min x d( 1); P = 0.11). When THI ranged between 72 and 84, nonriparian water availability did not have a significant impact on the amount of time cattle spent in the riparian zone or in riparian shade. These results suggest that water troughs placed away from unfenced streams may improve water quality by reducing the amount of time cattle spend in riparian zones when environmental conditions as evaluated by THI are not stressful. PMID- 19286819 TI - A comparative study on urinary purine derivative excretion of yak (Bos grunniens), cattle (Bos taurus), and crossbred (Bos taurus x Bos grunniens) in the Qinghai-Tibetan plateau, China. AB - Two experiments were conducted to determine the urinary purine derivative (PD) excretion rates and to analyze potential variations in PD excretions between yak, Tibetan indigenous cattle, and their crossbreds under similar ecological environment, and to develop prediction equations based on PD excretion for estimating rumen microbial protein supply. Three Tibetan castrated male yaks, and similar numbers of indigenous cattle and their crossbreds, were used in a fasting trial followed by a feeding trial, using three 3 x 3 Latin squares (1 for each breedtype), to measure responses of PD excretion to different feeding levels. The results showed that i) daily endogenous PD excretion for yak, indigenous cattle, and their crossbreds was 134, 163, and 138 micromol/kg of BW(0.75) (P = 0.38), respectively; and ii) crossbreds have greater PD excretion rate per unit digestible OM intake than indigenous cattle (P = 0.03). However, there were no significant differences between yaks and the crossbreds (P = 0.24) or between yaks and the indigenous cattle (P = 0.25). The proportion of allantoin to total PD in urine ranged from 0.83 to 0.88, with the crossbreds having greater values compared with the 2 parents (P = 0.03). Daily glomerular filtration rate, calculated using endogenous creatinine as an internal marker, for the above animals was 3.85, 4.23, and 3.61 L/kg of BW(0.75), respectively, in fasting trial (P = 0.59). The alpine animals may develop special regulating mechanisms in the kidney in terms of glomerular filtration rate and PD excretion, which would help the animals in adapting to the harsh environment of the Qinghai-Tibetan plateau. PMID- 19286820 TI - Intake and digestibility of naive kids differing in genetic resistance and experimentally parasitized (indoors) with Haemonchus contortus in two successive challenges. AB - We evaluated the effects of infection with Haemonchus contortus on feed intake, digestibility, fecal egg count, circulating eosinophils, and packed cell volume in Creole kids differing in genetic resistance (susceptible, S; resistant, R) to gastrointestinal parasitism and maintained on a similar level of nutrition. The experiment was carried out during 2 periods of 6 wk each differing in immunity development stage. In the first period (acquisition of immunity; period I), 22 naive male kids (23.4 +/- 0.65 kg of BW) were housed in individual boxes and fed a hay-based diet, and a primary infection was induced. In the second period (expression of immunity; period II), 15 of the initial 22 kids (28.4 +/- 0.77 kg of BW) were submitted to a secondary infection. Housing and management were uniform throughout the experiment. For each period, measurements of intake and digestibility were made at 0, 2, and 4 wk postinfection (WPI) with a single dose of 10,000 infective larvae (L(3)). The DMI and total-tract DM, OM, CP, NDF, and ADF digestibilities were determined using the total feces collection and ad libitum forage supply method. Fecal and blood samples were collected weekly to measure fecal egg count, circulating eosinophils, and packed cell volume. Infection with Haemonchus contortus decreased feed intake during period I. The absence of anorexia in period II was probably due to the acquired immunity of kids. The DMI was affected (P = 0.05) by genetic predisposition to resistance (626 vs. 583 +/- 26 g/d, for R vs. S) and WPI, being greatest in the second WPI (693 vs. 614 and 657 g/d, for WPI-2 vs. WPI-0 and WPI-4, respectively). The latter was related to worm establishment phase and was linked to the lower total tract digestibilities at this point. Digestibilities were least at WPI-2. The fecal egg counts were greater (P < 0.001) in period I than II, and differences between S and R were evident after the fifth WPI in period II. Circulating eosinophils were greater (P < 0.001) in S vs. R. The results suggest that effects of these parasites on intake and digestibility are influenced by the individual genetic resistance and the immunological stage, and the strongest impact occurs between the second and the third WPI, a period during which the immune response is more pronounced, probably due to parasite maturation. PMID- 19286821 TI - Effect of dietary conjugated linoleic acid supplementation in sows on performance and immunoglobulin concentration in piglets. AB - Conjugated linoleic acid isomers may affect the onset and severity of several diseases, including tumors, atherogenesis, and obesity. They may also modulate the immune response. However, little information regarding the most advantageous duration of CLA supplementation is available. The purpose of this study was to determine whether the length of dietary CLA supplementation of a sow affects growth, immune components, and metabolic and hormonal factors in lactating sows and piglets. Gestating sows were fed a control (0%) and a 0.5% CLA-supplemented diet beginning 7 d before parturition and ending 7 d postpartum (T1), or until weaning (T2; 7 sows per treatment). Colostrum and sow and piglet blood samples were collected for the determination of serum metabolite concentrations and immunoglobulin titer. Piglet BW at weaning were greater (P < 0.05) in the CLA groups compared with the control. Dietary CLA supplementation increased (P < 0.05) serum thyroxine concentration in sows, but serum insulin, glucose, NEFA, IGF-I, and leptin concentrations were not affected by CLA supplementation. Colostral IgG, IgA, and IgM titers were greater in sows fed CLA than in control sows (P < 0.05). At weaning (21 d), serum IgG titer of the piglets was greater (P < 0.05) in the T1 and T2 groups than the control group, but at 13 d postweaning, a difference (P < 0.05) was observed between the control and T2 group. The results from this study indicate potential beneficial effects of 0.5% dietary CLA supplementation from 7 d before parturition until 7 d postpartum in improving BW at weaning and immune components in piglets. PMID- 19286822 TI - Development and in vitro evaluation of an Escherichia coli probiotic able to inhibit the growth of pathogenic Escherichia coli K88. AB - Weaned piglets commonly suffer from gastroenteritis caused by enterotoxigenic Escherichia coli K88. Our aim was to produce E. coli strains that inhibited the growth of E. coli K88 and could be used as a probiotic against postweaning diarrhea. The inclusion criteria for the probiotics were that in addition to being able to inhibit E. coli K88, they also needed to be negative for virulence genes commonly associated with E. coli. A total of 463 E. coli isolates from the cattle rumen, cattle feces, swine feces, and soil were screened against 18 E. coli K88 clinical isolates using an agar diffusion technique. Growth inhibition of the most sensitive K88 indicator strain 2-12 occurred for 121/463 isolates: 96/358 from cattle feces, 0/33 from rumen fluid, 9/35 from swine feces, and 16/37 from soil. Of the 121 positive strains, 71/121 were negative for toxin genes (LT, STa, STb, VT1, and VT2). The 14 most inhibitory strains were screened against a range of substrates to assess the ability to utilize carbohydrates that could be included in the diet to enhance their ability to compete in the gut. Two strains, UM-2 and UM-7, were weak utilizers of starch and inulin. In vitro competition assays between the probiotic strains and E. coli K88 strain 2-12 were conducted with glucose as the only carbon source (minimal medium; MM), MM + 2% starch, or MM + 2% inulin. The UM-2 and UM-7 strains were able to outcompete strain 2-12 when glucose was the only carbon source, indicating that inhibitory activity was produced against 2-12 independent of carbon source. The UM-2 strain outcompeted strain 2-12 in assays in which potato starch or inulin was the only carbon source; the ability of 2-12 to maintain its concentrations in the culture were probably the result of cross feeding of breakdown sugars of starch and inulin that could be utilized by 2-12. In contrast, UM-7 did not grow as well as UM-2 on starch and inulin and 2-12 declined rapidly in successive cultures likely because of the lack of breakdown products of starch and inulin produced by UM-7. We conclude that probiotic E. coli without known toxins and that produce inhibitory activity against E. coli that cause postweaning diarrhea can be produced. In addition, the ability to utilize starch or inulin is an important phenotype because it likely gives the probiotic a competitive advantage in the gut. PMID- 19286823 TI - Development of estrous synchronization protocols using melengestrol acetate in Bos indicus cattle. AB - Five experiments were conducted on commercial farms in Brazil designed to develop the basis for an estrus synchronization protocol using melengestrol acetate (MGA) in Bos indicus cattle. These studies resulted in the development of the following protocol: 0.5 mg x d(-1) of MGA between d -14 and -1; 2.0 mg i.m. injection of estradiol cypionate on d -9; 48 h temporary weaning between d 0 and 2; and natural service beginning on d 0. The basis of this protocol was to induce estrous cyclicity before postpartum loss of body condition, prevent premature luteolysis, eliminate the need for labor required to detect estrus, and consequently increase the likelihood of pregnancy early during the postpartum period. This treatment effectively induced estrous cyclicity among anestrous cows, synchronized estrus activity, and prevented premature luteolysis with no negative effect on pregnancy. PMID- 19286824 TI - Substrate utilization and dose response to insulin by subcutaneous adipose tissue of Angus steers fed corn- or hay-based diets. AB - We hypothesized that, at a common age endpoint, adipose tissue from corn-fed steers would be less sensitive to insulin than adipose tissue from hay-fed steers. Angus steers were assigned to either a corn-based diet (n = 6) or hay based diet (n = 6) and fed to common days on feed. Steers fed the corn-based diet had 2.44 cm of fat thickness over the 12th thoracic rib, whereas hay-fed steers had 1.04 cm of fat thickness. At slaughter, subcutaneous adipose samples were collected and portions of subcutaneous adipose tissue were incubated with [U (14)C]acetate to quantify fatty acid synthesis or with [U-(14)C]glucose to assess glucose utilization in the presence of 0, 100, or 500 ng/mL of insulin. Additional subcutaneous samples were used to evaluate glycolytic intermediate concentrations as indicators of glycolytic flux. Data were analyzed as a split plot with diet in the main plot and insulin concentration and its interaction with diet in the sub-plot. Within diet, linear and quadratic contrasts of insulin concentration were tested. Diet had no effect (P > or = 0.31) on glucose metabolism or acetate carbon incorporation into total lipids (P = 0.32). Insulin had no effect (P > 0.21) on glucose conversion to CO(2), lactate, or total lipids, nor did it affect (P = 0.28) acetate conversion to total lipids. No diet x insulin interaction (P > 0.36) was observed for any measure of subcutaneous metabolism in vitro. Steers fed the corn-based diet exhibited neither a linear (P > 0.22) nor a quadratic (P > 0.24) effect to increasing insulin concentration. However, when steers were fed the hay-based diet, there was a positive linear (P = 0.06) effect for glucose oxidation. These results suggest that subcutaneous adipose tissue may become resistant to stimulation by insulin in steers fed to a fat thickness above the average feedlot steer, but this is independent of diet. PMID- 19286825 TI - Effects of colostrum replacer supplemented with lactoferrin on the blood plasma immunoglobulin G concentration and intestinal absorption of xylose in the neonatal calf. AB - The objective of this experiment was to determine the effect of lactoferrin (Lf) supplementation of colostrum replacer (CR) fed as 1 or 2 doses on apparent efficiency of IgG absorption, serum IgG concentrations, and xylose absorption/intestinal development in neonatal Holstein bull calves. Eighty bull calves were assigned to a 2 x 4 factorial arrangement of treatments in a randomized complete block design. Calves used were from the University of New Hampshire (n = 48) or a commercial dairy (n = 32). All calves were fed CR according to manufacturer's recommendations +/- Lf treatment within 90 min of birth. Lactoferrin treatments were 0 (control), 0.5, 1, and 2 g/d of supplemental Lf. At 12 h of age, calves were fed a second feeding of CR +/- supplemental Lf or 2 L of milk replacer +/- supplemental Lf. Calves continued to be fed milk replacer +/- supplemental Lf every 12 h for the duration of study. Blood samples were collected for determination of IgG and total serum protein at 0, 6, 12, 18, 24, and 48 h. Calves, except those fed 1 dose of CR plus 1 or 2 g of Lf, had serum IgG concentrations > or = 10 g/L at 24 h. Lactoferrin supplementation had no effect on serum IgG or total serum protein concentrations. Calves fed 2 doses of CR had greater serum IgG concentrations compared with calves fed 1 dose of CR. Apparent efficiency of absorption of IgG was less in calves fed 2 doses of CR compared with calves fed 1 dose of CR. Lactoferrin supplementation (up to 1 g/d) resulted in decreased apparent efficiency of absorption of IgG. At the sixth feeding (60 +/- 2 h of age), D-xylose (0.5 g/kg BW) was mixed with milk replacer +/- supplemental Lf (n = 48) to determine xylose absorption by the small intestine. Blood was collected at 0, 2, 4, 6, 8, and 12 h after feeding xylose for determination of plasma glucose and xylose concentrations. Xylose means and area under the curve resulted in quadratic effects. Feeding calves 0.5 or 1 g/d supplemental Lf resulted in decreased plasma xylose concentrations compared with calves fed 0 or 2 g/d of supplemental Lf. Colostrum replacer or supplemental Lf did not affect plasma glucose concentrations. This study indicates that supplementing Lf at 0.5 or 1 g/d to calves fed CR has a negative effect on apparent efficiency of IgG absorption and xylose absorption. PMID- 19286826 TI - Evaluation of antral follicle count and ovarian morphology in crossbred beef cows: investigation of influence of stage of the estrous cycle, age, and birth weight. AB - Depletion of the ovarian reserve is associated with reproductive senescence in mammalian females, and there is a positive relationship between the size of the ovarian reserve and the number of antral follicles on the surface of the ovary. Therefore, we conducted a series of experiments to investigate the influence of stage of the estrous cycle, age, and birth weight on antral follicle counts (AFC) in beef cows and heifers. Pairs of ovaries were collected from crossbred beef cows at slaughter (n = 72) or at necropsy (n = 333; 0 to 11 yr of age); all visible antral follicles were counted, the ovaries were weighed, and stage of the estrous cycle was estimated based on ovarian morphology. There was no influence of estimated stage of the estrous cycle on AFC (P = 0.36). There was a small but positive effect of birth weight on AFC [AFC = -1.7 + 0.31(birth weight); P = 0.007, r(2) = 0.05]. When antral follicle counts were regressed on age, there was a quadratic effect of age such that AFC increased until 5 yr of age and decreased thereafter [AFC = 12.9 + 9.0(yr) - 0.86(yr(2)); P < 0.001, r(2) = 0.22]. In a third experiment, crossbred beef heifers (n = 406; 353 to 463 d of age) at 3 locations were subjected to ovarian ultrasonography on unknown day of the estrous cycle. Heifers were classified as low AFC (<15 follicle, n = 84) or high AFC (>24 follicles, n = 178). Whereas estimated stage of the estrous cycle did not influence AFC (P = 0.62), heifers classified as low AFC had smaller ovaries (P = 0.001), decreased birth weight (P = 0.003), and a decreased heifer pregnancy rate (P = 0.05) compared with heifers in the high AFC group. From these results, we conclude that AFC in beef cows and heifers is influenced by birth weight and age but not by stage of the estrous cycle. In beef cows, the number of antral follicles increases to 5 yr of age and then begins to decline. This may indicate that a decrease in fertility due to decline of the ovarian reserve may begin earlier than previously thought in beef cows. PMID- 19286827 TI - Amino acid digestibility and concentration of digestible and metabolizable energy in soybean meal produced from conventional, high-protein, or low-oligosaccharide varieties of soybeans and fed to growing pigs. AB - Two experiments were conducted to determine AA digestibility and the concentration of DE and ME in 5 sources of soybean meal (SBM). The 5 sources included hexane-extracted SBM produced from high-protein soybeans (SBM-HP) and conventional soybeans (SBM-CONV), and mechanically extruded-expelled SBM produced from high-protein soybeans (EE-SBM-HP), low-oligosaccharide soybeans (EE-SBM-LO), and conventional soybeans (EE-SBM-CONV). Five diets that each contained 1 source of SBM and a N-free diet were used in Exp. 1 to determine AA digestibility in each meal. Twelve growing barrows (initial BW: 67.7 +/- 1.34 kg) were allotted to a replicated 6 x 6 Latin square design with 6 periods and 6 diets in each square. Each period lasted 7 d, and ileal digesta were collected on d 6 and 7 of each period. Results of the experiment showed that the standardized ileal digestibility (SID) of all AA except Trp was similar for SBM-HP and SBM-CONV, but EE-SBM-HP and EE-SBM-LO had greater (P < 0.05) SID of His, Ile, Lys, Thr, and Val than EE-SBM-CONV. The SID of all indispensable AA in EE-SBM-HP was greater (P < 0.05) than in SBM-HP. The SID of Arg, Ile, Leu, and Phe in EE-SBM-CONV was greater (P < 0.05) than in SBM-CONV, but the SID of Trp was also greater (P < 0.05) in SBM-CONV than in EE-SBM-CONV. Experiment 2 was conducted to measure DE and ME in the same 5 sources of SBM as used in Exp. 1. Forty-eight growing barrows (initial BW: 38.6 +/- 3.46 kg) were placed in metabolism cages and randomly allotted to 6 diets with 8 replicates per diet. A corn-based diet and 5 diets based on a mixture of corn and each source of SBM were formulated. Urine and feces were collected during a 5-d collection period, and values for DE and ME in each source of SBM were calculated using the difference procedure. Results showed that the ME in SBM-HP tended to be greater (P = 0.10) than in SBM-CONV (4,074 vs. 3,672 kcal/kg of DM). The ME in EE-SBM-HP also tended to be greater (P = 0.10) than in EE-SBM-CONV and in EE-SBM-LO (4,069 vs. 3,620 and 3,721 kcal/kg of DM), but there was no difference in ME between extracted and extruded-expelled meals. It is concluded that SBM-HP has a greater feeding value than SBM-CONV because of greater concentrations of digestible AA and ME. Likewise, EE-SBM-LO has a greater concentration of most indispensable AA than EE-SBM-CONV, but the concentration of ME is similar in these 2 meals. Results of this experiment also showed that AA digestibility values in extruded-expelled SBM are greater than in hexane-extracted SBM. PMID- 19286829 TI - Development of the Korean primary care assessment tool--measuring user experience: tests of data quality and measurement performance. AB - OBJECTIVE: To develop a tool for assessing the performance of primary care services in South Korea from the patient's perspective and to test the validity of the tool under the conceptual framework of the recently developed definition of primary care in Korea. DESIGN: Item development for questionnaire and a cross sectional survey for tool validation at 16 primary care clinics. All family physicians included in this study were required to have practiced at their current clinic for at least 2 years. PARTICIPANTS: A nine expert panel was assembled for tool development and patients (or guardians) who had visited their primary care clinic on six or more occasions over a period of more than 6 months participated in the survey. MAIN OUTCOME MEASURES: Scores corresponding to each domain of primary care. RESULTS: A total of 722 effective data sets were used for the analysis. Five items were eliminated from the preliminary 30-item tool after expert discussions at two seminars. Another four items were eliminated by principle component analysis. For each of the four domains (comprehensiveness, coordination function, personalized care, and family/community orientation), tests of scaling assumptions were well satisfied by all Likert-scaled measures. On the other hand, 'first contact' turned out to be a composite domain with five independent single-item scales. CONCLUSION: The Korean primary care assessment tool (version 1) consists of four multi-item scales and one composite scale. Widespread application of this tool will provide an empirical basis for the measurement, monitoring and continuous improvement of primary care in South Korea. PMID- 19286830 TI - Evolutionary construction of multiple graph alignments for the structural analysis of biomolecules. AB - The concept of multiple graph alignment (MGA) has recently been introduced as a novel method for the structural analysis of biomolecules. Using approximate graph matching techniques, this method enables the robust identification of approximately conserved patterns in biologically related structures. In particular, MGA enables the characterization of functional protein families independent of sequence or fold homology. This article first recalls the concept of MGA and then addresses the problem of computing optimal alignments from an algorithmic point of view. In this regard, a method from the field of evolutionary algorithms is proposed and empirically compared with a hitherto existing heuristic approach. Empirically, it is shown that the former yields significantly better results than the latter, albeit at the cost of an increased runtime. PMID- 19286831 TI - Site of metabolism prediction for six biotransformations mediated by cytochromes P450. AB - MOTIVATION: One goal of metabolomics is to define and monitor the entire metabolite complement of a cell, while it is still far from reach since systematic and rapid approaches for determining the biotransformations of newly discovered metabolites are lacking. For drug development, such metabolic biotransformation of a new chemical entity (NCE) is of more interest because it may profoundly affect its bioavailability, activity and toxicity profile. The use of in silico methods to predict the site of metabolism (SOM) in phase I cytochromes P450-mediated reactions is usually a starting point of metabolic pathway studies, which may also assist in the process of drug/lead optimization. RESULTS: This article reports the Cytochromes P450 (CYP450)-mediated SOM prediction for the six most important metabolic reactions by incorporating the use of machine learning and semi-empirical quantum chemical calculations. Non local models were developed on the basis of a large dataset comprising 1858 metabolic reactions extracted from 1034 heterogeneous chemicals. For validation, the overall accuracies of all six reaction types are higher than 0.81, four of which exceed 0.90. In further receiver operating characteristic (ROC) analyses, each of the SOM model gave a significant area under curve (AUC) value over 0.86, indicating a good predicting power. An external test was made on a previously published dataset, of which 80% of the experimentally observed SOMs can be correctly identified by applying the full set of our SOM models. AVAILABILITY: The program package SOME_v1.0 (Site Of Metabolism Estimator) developed based on our models is available at http://www.dddc.ac.cn/adme/myzheng/SOME_1_0.tar.gz. PMID- 19286832 TI - A novel parallel approach to the likelihood-based estimation of admixture in population genetics. AB - SUMMARY: Inferring population admixture from genetic data and quantifying it is a difficult but crucial task in evolutionary and conservation biology. Unfortunately state-of-the-art probabilistic approaches are computationally demanding. Effectively exploiting the computational power of modern multiprocessor systems can thus have a positive impact to Monte Carlo-based simulation of admixture modeling. A novel parallel approach is briefly described and promising results on its message passing interface (MPI)-based C++ implementation are reported. AVAILABILITY: The software package parLEA is freely available at (http://dm.unife.it/parlea). PMID- 19286833 TI - Statistical detection of cooperative transcription factors with similarity adjustment. AB - MOTIVATION: Statistical assessment of cis-regulatory modules (CRMs) is a crucial task in computational biology. Usually, one concludes from exceptional co occurrences of DNA motifs that the corresponding transcription factors (TFs) are cooperative. However, similar DNA motifs tend to co-occur in random sequences due to high probability of overlapping occurrences. Therefore, it is important to consider similarity of DNA motifs in the statistical assessment. RESULTS: Based on previous work, we propose to adjust the window size for co-occurrence detection. Using the derived approximation, one obtains different window sizes for different sets of DNA motifs depending on their similarities. This ensures that the probability of co-occurrences in random sequences are equal. Applying the approach to selected similar and dissimilar DNA motifs from human TFs shows the necessity of adjustment and confirms the accuracy of the approximation by comparison to simulated data. Furthermore, it becomes clear that approaches ignoring similarities strongly underestimate P-values for cooperativity of TFs with similar DNA motifs. In addition, the approach is extended to deal with overlapping windows. We derive Chen-Stein error bounds for the approximation. Comparing the error bounds for similar and dissimilar DNA motifs shows that the approximation for similar DNA motifs yields large bounds. Hence, one has to be careful using overlapping windows. Based on the error bounds, one can precompute the approximation errors and select an appropriate overlap scheme before running the analysis. AVAILABILITY: Software to perform the calculation for pairs of position frequency matrices (PFMs) is available at http://mosta.molgen.mpg.de as well as C++ source code for downloading. PMID- 19286834 TI - SimMLST: simulation of multi-locus sequence typing data under a neutral model. AB - SUMMARY: Multi-locus sequence typing (MLST) is a widely used method of characterization of bacterial isolates. It has been applied to over 50,000 isolates in over 50 different species. Here, we present a coalescent method to jointly simulate MLST data and the clonal genealogy that gave rise to the sample. AVAILABILITY AND IMPLEMENTATION: SimMLST was implemented in C++ and Qt4 for the graphical user interface. It is distributed under the terms of the GNU General Public License. Source code and binaries for Windows and Linux are available from (http://go.warwick.ac.uk/SimMLST). A user guide and a technical description of the algorithm are provided with the program. PMID- 19286835 TI - Activation of Rho and Rho-associated kinase by GPR54 and KiSS1 metastasis suppressor gene product induces changes of cell morphology and contributes to apoptosis. AB - The mechanism of action of the metastasis suppressor KiSS1 and its receptor GPR54 is still incompletely characterized. Although the loss of KiSS1 expression by tumor cells has been associated with a metastatic phenotype, the nature of the cellular target of the secreted kisspeptins is unknown. Although an autocrine model of action has been generally assumed, metastasis suppression by KiSS1 has also been shown in cells that do not express GPR54, suggesting a paracrine mechanism in which kisspeptins affect cells in the metastatic niche. Activation of GPR54 was shown to inhibit cell motility and invasion of tumor cells, induce the formation of stress fibers, and reduce the expression of matrix metalloproteinase 9. We showed previously that the activation of GPR54 by kisspeptin-10 suppressed CXCR4-mediated chemotaxis in response to stromal cell derived factor 1/CXCL12 and abolished the phosphorylation of Akt by CXCR4. We also demonstrated that activation of GPR54 inhibited Akt phosphorylation after the activation of epidermal growth factor receptor and the insulin receptor and triggered apoptosis in epithelial and lymphoid cell lines through a mechanism involving extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase. We show here that the activation of GPR54 induced immediate and profound changes of cell morphology, including cytoplasmic condensation and formation of unpolarized plasma membrane protrusions. These events were dependent on Rho and Rho-Associated Kinase (ROCK) activation. The activation of ROCK also contributed to GPR54-mediated apoptosis in 293 cells, and its effect was additive to and independent of ERK activation. These results suggest that RhoA and ROCK are additional key components of the antimetastatic effect of kisspeptins. PMID- 19286836 TI - Regulation of human vitamin D(3) 25-hydroxylases in dermal fibroblasts and prostate cancer LNCaP cells. AB - In this study, we examined whether 1alpha,25-dihydroxyvitamin D(3) (calcitriol), phenobarbital, and the antiretroviral drug efavirenz, drugs used by patient groups with high incidence of low bone mineral density, could affect the 25 hydroxylase activity or expression of human 25-hydroxylases in dermal fibroblasts and prostate cancer LNCaP cells. Fibroblasts express the 25-hydroxylating enzymes CYP2R1 and CYP27A1. LNCaP cells were found to express two potential vitamin D 25 hydroxylases-CYP2R1 and CYP2J2. The presence in different cells of nuclear receptors vitamin D receptor (VDR), pregnane X receptor (PXR), and constitutive androstane receptor (CAR) was also determined. Phenobarbital suppressed the expression of CYP2R1 in fibroblasts and CYP2J2 in LNCaP cells. Efavirenz suppressed the expression of CYP2R1 in fibroblasts but not in LNCaP cells. CYP2J2 was slightly suppressed by efavirenz, whereas CYP27A1 was not affected by any of the two drugs. Calcitriol suppressed the expression of CYP2R1 in both fibroblasts and LNCaP cells but had no clear effect on the expression of either CYP2J2 or CYP27A1. The vitamin D(3) 25-hydroxylase activity in fibroblasts was suppressed by both calcitriol and efavirenz. In LNCaP cells, consumption of substrate (1alpha-hydroxyvitamin D(3)) was used as indicator of metabolism because no 1alpha,25-dihydroxyvitamin D(3) product could be determined. The amount of 1alpha hydroxyvitamin D(3) remaining in cells treated with calcitriol was significantly increased. Taken together, 25-hydroxylation of vitamin D(3) was suppressed by calcitriol and drugs. The present study provides new information indicating that 25-hydroxylation of vitamin D(3) may be regulated. In addition, the current results may offer a possible explanation for the impaired bone health after treatment with certain drugs. PMID- 19286837 TI - The epidemiological patterns of honour killing of women in Pakistan. AB - BACKGROUND: Honour killing (HK) is a problem of public health concern but published data on the phenomenon are limited and many cases likely go unrecognized. Our study focuses on the epidemiological patterns of HK of women in Pakistan, where domestic violence is common and HK occurs but is poorly described. METHODS: Human Rights Commission of Pakistan (HRCP) systematically collected data on HK of women using newspaper reports from January 2004 till December 2007. We analysed the aggregated data on HK through December 2007 and estimated the rates of HK. RESULTS: A total of 1957 HK events occurred from 2004 to 2007; complete data was not available for all variables. Adults (>or=18 years) constituted 82% (803/978) of death toll with 88% (1257/1435) being married. Alleged extramarital relation was the major reason for the killing (92%; 1759/1902). Husbands (43%; 749/1739), brothers (24%; 421/1739) and 'other' close relatives (12%; 200/1739) were the perpetrators in known HK events. Among the weapons/methods used for killing, firearms (61%; 1071/1768), stabbing (4%; 65/1768), use of axe (12%; 220/1768), edged tool (8%;136/1768) and strangulation (9%; 167/1768) were the main means of execution. The mean annual rate of HK in females (age 15-64 years) was found to be 15.0 per million. CONCLUSIONS: Newspaper reports are good source of surveillance when information is limited. We found that adult married women constituted the majority of victims of HK. Ongoing surveillance would serve to better characterize HK in Pakistan and assess the effectiveness of preventive strategies. PMID- 19286838 TI - Age and closeness of death as determinants of health and social care utilization: a case-control study. AB - BACKGROUND: We used case-control design to compare utilization of health and social services between older decedents and survivors, and to identify the respective impact of age and closeness of death on the utilization of services. METHODS: Data were derived from multiple national registers. The sample consisted of 56,001 persons, who died during years 1998-2000 at the age of > or = 70, and their pairs matched on age, gender and municipality of residence, who were alive at least 2 years after their counterpart's death. Data include use of hospitals, long-term care and home care. Decedents' utilization within 2 years before death and survivors' utilization in the same period of time was assessed in three age groups (70-79, 80-89 and > or = 90 years) and by gender. RESULTS: Decedents used hospital and long-term care more than their surviving counterparts, but the time patterns were different. In hospital care the differences between decedents and survivors rose in the last months of the study period, whereas in long-term care there were clear differences during the whole 2-year period. The differences were smaller in the oldest age group than in younger age groups. CONCLUSION: Closeness of death is an important predictor of health and social service use in old age, but its influence varies between age groups. Not only the changing age structure, but also the higher average age at death affects the future need for services. PMID- 19286839 TI - Effect of a 24+ hour fast on breast milk composition. AB - In this preliminary prospective study, breast milk is sampled surrounding 4 religious fast days to determine the effect of a more than 24-hour fast on breast milk composition. The participants are 48 healthy women nursing healthy babies between 1 and 6 months of age. Samples are collected within 2 days before the fast (baseline), immediately after the fast, and 24 hours after fast completion. Samples are tested for sodium, calcium, phosphorus, triglycerides, total protein, and lactose. From baseline to immediately after fast, mean sodium, calcium, and protein levels increase (P = .013, P < .0001, and P < .0001, respectively) and mean phosphorus and lactose levels decrease (P < .0001 and P = .003, respectively). Mean triglycerides are unchanged. Twenty-four hours after fast, parameters are no longer significantly different from baseline except for elevated mean protein levels (P = .022) and lactose that is still reduced (P = .017). A fast of this nature is statistically associated with certain biochemical changes in breast milk. PMID- 19286840 TI - US national breastfeeding monitoring and surveillance: current status and recommendations. AB - Eleven federally funded datasets assessing breastfeeding behaviors in the United States (Early Childhood Longitudinal Survey, Infant Feeding Practices Survey II, National Health and Nutrition Examination Survey, National Immunization Survey, National Survey of Children's Health, National Survey of Early Childhood Health, National Survey of Family Growth, Pediatric Nutrition Surveillance System, Pregnancy Nutrition Surveillance System, Pregnancy Risk Assessment Monitoring Survey, and WIC Participant and Program Characteristics) were reviewed to evaluate breastfeeding variables (initiation, duration and exclusivity) and determine whether relevant breastfeeding determinants were collected to evaluate breastfeeding practices from a health disparities perspective. The datasets used inconsistent breastfeeding definitions, limited ethnic descriptors, and varied regarding availability of relevant determinants. Multiple datasets collect breastfeeding data, but a coordinated US breastfeeding monitoring and surveillance system does not exist. Suggestions to improve this system include: standardizing breastfeeding definitions, expanding ethnic/racial descriptors, collecting additional relevant variables, and reducing recall periods. PMID- 19286841 TI - Barriers to best outcomes in breastfeeding for Maori: mothers' perceptions, whanau perceptions, and services. AB - This research explores the perceptions of New Zealand Maori women and their whanau (customary Maori extended family) toward barriers to achieving best outcomes in infant feeding: exclusively breastfed infants at 6 months. Interviews are undertaken with 59 Maori women who have given birth in the previous 3 years and 27 whanau members. Although mothers and whanau members feel positively toward breastfeeding and generally expect to breastfeed exclusively, these expectations are unmet in many cases because of lack of support when establishing breastfeeding; lack of support when life circumstances change; lack of timely, culturally relevant, and comprehensible information; confusion about smoking while breastfeeding; uncertainty about the safety of bed-sharing, and perceived lack of acceptability of breastfeeding in public. The relatively high rates of tobacco use by Maori create a tension for breastfeeding mothers, cited by some as a reason for ending breastfeeding prematurely. PMID- 19286842 TI - Fentanyl transdermal analgesia during pregnancy and lactation. AB - This report describes an infant who was born to a mother with chronic pain treated with fentanyl 100 microg/h transdermal patch throughout her pregnancy and during lactation. On day of life 27, when the baby was feeding and gaining weight on maternal milk, samples of the baby's blood and maternal milk were sent for analysis. The mother's milk fentanyl level was 6.4 ng/ mL. The infant's blood fentanyl level was undetectable. This preliminary report suggests that fentanyl transdermal patch treatment might be a viable option for managing chronic pain during lactation. PMID- 19286843 TI - Polymorphisms of DNA damage response genes in radiation-related and sporadic papillary thyroid carcinoma. AB - Papillary thyroid carcinoma (PTC) etiologically occurs as a radiation-induced or sporadic malignancy. Genetic factors contributing to the susceptibility to either form remain unknown. In this retrospective case-control study, we evaluated possible associations between single-nucleotide polymorphisms (SNPs) in the candidate DNA damage response genes (ATM, XRCC1, TP53, XRCC3, MTF1) and risk of radiation-induced and sporadic PTC. A total of 255 PTC cases (123 Chernobyl radiation-induced and 132 sporadic, all in Caucasians) and 596 healthy controls (198 residents of Chernobyl areas and 398 subjects without history of radiation exposure, all Caucasians) were genotyped. The risk of PTC and SNPs interactions with radiation exposure were assessed by logistic regressions. The ATM G5557A and XRCC1 Arg399Gln polymorphisms, regardless of radiation exposure, associated with a decreased risk of PTC according to the multiplicative and dominant models of inheritance (odds ratio (OR) = 0.69, 95% confidence interval (CI) 0.45-0.86 and OR = 0.70, 95% CI 0.59-0.93 respectively). The ATM IVS22-77 T > C and TP53 Arg72Pro SNPs interacted with radiation (P = 0.04 and P = 0.01 respectively). ATM IVS22-77 associated with the increased risk of sporadic PTC (OR = 1.84, 95% CI 1.10-3.24) whereas TP53 Arg72Pro correlated with the higher risk of radiogenic PTC (OR = 1.80, 95% CI 1.06-2.36). In the analyses of ATM/TP53 (rs1801516/rs664677/rs609429/rs1042522) combinations, the GG/TC/CG/GC genotype strongly associated with radiation-induced PTC (OR = 2.10, 95% CI 1.17-3.78). The GG/CC/GG/GG genotype displayed a significantly increased risk for sporadic PTC (OR = 3.32, 95% CI 1.57-6.99). The results indicate that polymorphisms of DNA damage response genes may be potential risk modifiers of ionizing radiation induced or sporadic PTCs. PMID- 19286844 TI - Plasma 25,OH vitamin D concentrations are not associated with rheumatoid arthritis (RA)-related autoantibodies in individuals at elevated risk for RA. AB - OBJECTIVE: To evaluate the association between rheumatoid arthritis (RA)-related autoantibodies and plasma 25,OH vitamin D in subjects at risk for RA. METHODS: In 1210 subjects without RA, 76 were positive for anti-cyclic citrullinated peptide antibodies or for at least 2 rheumatoid factors (RF; by nephelometry: RF-IgM, RF IgG, RF-IgA). 25,OH vitamin D was measured in these cases and 154 autoantibody negative controls from this cohort. RESULTS: 25,OH vitamin D levels did not differ between cases and controls (adjusted OR 1.23, 95% CI 0.93-1.63). CONCLUSION: Vitamin D concentrations are not associated with RA-related autoimmunity in unaffected subjects at increased risk for RA. PMID- 19286845 TI - Structural polymorphisms in the mannose-binding lectin gene are associated with juvenile idiopathic arthritis. AB - OBJECTIVE: To investigate the possible association between polymorphisms of the mannose-binding lectin gene (MBL2) and susceptibility to juvenile idiopathic arthritis (JIA). METHODS: We performed a case-control association study including 118 Hungarian patients with JIA and 118 sex-matched healthy controls. MBL genotyping for the 3 mutant structural alleles at codons 54 (B), 57 (C), and 52 (D) in exon 1 and the promoter polymorphisms at position -550 (HL) and -221 (YX) were carried out by real-time PCR allelic discrimination. Serum level of MBL was determined by ELISA. RESULTS: Variant allele frequencies of both codon 52 and 57 polymorphisms in the MBL2 gene were significantly overrepresented in JIA (p=0.001 and p=0.004, respectively). The frequency of low MBL genotypes (XA/XA, YA/YO, XA/YO, and YO/YO) in JIA was higher than that in healthy controls (p=0.001). Serum MBL concentrations were found to be significantly lower in JIA patients versus control subjects (p=0.001). The 2 promoter polymorphisms and codon 54 SNP of the MBL2 gene were not associated with JIA. CONCLUSION: Our findings suggest that genetically determined low MBL levels may predispose children to JIA in a Hungarian population. These data warrant further research to investigate the role of the lectin-dependent complement system in the pathogenesis of JIA. PMID- 19286846 TI - Anti-cyclic citrullinated peptide antibodies distinguish hepatitis B virus (HBV) associated arthropathy from concomitant rheumatoid arthritis in patients with chronic HBV infection. AB - OBJECTIVE: To determine whether anti-cyclic citrullinated peptide (anti-CCP) antibodies, which are a highly specific test for rheumatoid arthritis (RA), could differentiate between hepatitis B virus (HBV)-associated arthropathy and concomitant RA in Korean patients with chronic HBV infection. METHODS: We investigated 240 patients with HBV infection. Anti-CCP antibodies were measured by ELISA and rheumatoid factor (RF) by the latex fixation test. Patient records were reviewed, and a standard form was used to record all demographic, clinical, and laboratory characteristics. Patients were divided into 4 groups according to joint symptoms: asymptomatic, arthralgia, oligoarthritis, and RA. We categorized liver disease into 3 groups: carrier, chronic hepatitis, and cirrhosis. RESULTS: Anti-CCP antibodies and RF were detected in 11 and 28 of 240 patients, respectively. Anti-CCP antibodies were detected in 9 of 10 RA (90%) and 2 of 230 non-RA patients (0.86%). The positive rate for RF was 90% in RA and 8.3% in non RA. Eight of 10 RA patients were positive for both RF and anti-CCP antibodies. RF was detected in 11 patients without joint symptoms, 4 with arthralgia, and 4 with oligoarthritis, whereas anti-CCP antibodies were found in 1 patient without joint symptoms and 1 with oligoarthritis. Specificity of anti-CCP antibody for RA was 99.1%, whereas RF showed 91.7% specificity (p<0.0002). We compared the titers and positive detection rates of anti-CCP antibodies and RF among liver disease subgroups. There was no significant between-subgroup difference. CONCLUSION: Measurement of anti-CCP antibodies is better than RF detection to discriminate HBV-associated arthropathy from concomitant RA in patients with chronic HBV infection. PMID- 19286847 TI - Clinical efficacy and safety of successful longterm urate lowering with febuxostat or allopurinol in subjects with gout. AB - OBJECTIVE: To determine longterm urate-lowering efficacy and clinical benefits and safety of therapy with febuxostat or allopurinol in subjects with gout. METHODS: Subjects (n=1086) in this open-label extension study were assigned to fixed-dose daily urate-lowering treatment (ULT) with febuxostat (80 mg or 120 mg) or allopurinol (300 mg). ULT reassignment was permitted during months 1 to 6 to achieve serum urate (SUA) concentrations between 3.0 and <6.0 mg/dl. Flares requiring treatment, tophus size, safety, and SUA levels were monitored during up to 40 months of ULT maintenance. RESULTS: After 1 month initial treatment, >80% of subjects receiving either febuxostat dose, but only 46% of subjects receiving allopurinol, achieved SUA<6.0 mg/dl. After ULT reassignment, >80% of all remaining subjects maintained the primary efficacy endpoint of SUA<6.0 mg/dl at each visit. More subjects initially randomized to allopurinol required ULT reassignment to achieve SUA<6.0 mg/dl compared with subjects receiving febuxostat. Maintenance of SUA<6.0 mg/dl resulted in progressive reduction to nearly 0 in proportion of subjects requiring gout flare treatment. Baseline tophus resolution was achieved by 46%, 36%, and 29% of subjects maintained on febuxostat 80 mg, febuxostat 120 mg, and allopurinol, respectively. Overall adverse event rates (including cardiovascular adverse event rates), adjusted for 10-fold greater febuxostat than allopurinol exposure, did not differ significantly among treatment groups. CONCLUSION: Durable maintenance of goal range SUA level with either dose of febuxostat or in smaller numbers of subjects with allopurinol resulted in near elimination of gout flares and improved tophus status over time. Registered as NCT00175019. PMID- 19286848 TI - Detection of unsaturated disaccharides, pyridinoline, and hydroxyproline in urine of patients with Kashin-Beck disease: comparison with controls in an endemic area. AB - OBJECTIVE: To investigate the pathologic status of adult patients with Kashin Beck disease (KBD) in an endemic area of China through detection of 5 biochemical markers in their urine, and to study the correlations between these markers and KBD. METHODS: A total of 55 patients with KBD over age 40 years were recruited and divided into groups, Grade 1 and Grade 2, according to clinical diagnosis criteria for KBD and our inclusion criteria; 25 healthy persons were enrolled into a control group. The first-time urine of the 80 participants was collected in the morning. Three unsaturated disaccharides, pyridinoline (PYD), and hydroxyproline (HYP) were detected in urine samples with high performance liquid chromatography, ELISA, and a chemical kit. Mean levels of these markers were compared in the 3 groups. RESULTS: The mean concentrations of 3 unsaturated disaccharides and PYD in the Grade 2 group were significantly higher than levels in the Grade 1 group and controls (p<0.05). There was no significant difference between findings in the Grade 1 group and controls. Levels of 3 unsaturated disaccharides correlated with each other (p<0.01). The correlation coefficient between PYD and HYP was 0.470 (p<0.01). Except for HYP, the other markers all correlated with grade of KBD, rather than age or sex of subjects. CONCLUSION: The cartilage degradation of patients with Grade 2 KBD was more severe than that of Grade 1 patients and controls. The pathologic condition of Grade 1 patients was mild. Except for HYP, the markers we investigated specifically reflected the pathologic bone metabolism of adult patients with KBD. Trial registration number ChiCTR-TRC-00000140. PMID- 19286849 TI - Surfactant protein D and KL-6 as serum biomarkers of interstitial lung disease in patients with scleroderma. AB - OBJECTIVE: To assess whether serum concentrations of surfactant protein D (SP-D) and Krebs von den Lungen-6 (KL-6), glycoproteins expressed by type II pneumocytes, correlate with the presence of "alveolitis" and measures of lung function in patients enrolled in the Scleroderma Lung Study (SLS). METHODS: Serum obtained at baseline screening of patients with systemic sclerosis (SSc, scleroderma) in the SLS was assayed. "Alveolitis" was defined by either bronchoalveolar lavage or thoracic high-resolution computed tomography (HRCT) by SLS criteria. SP-D and KL-6 levels were measured by ELISA in 66 SSc patients (44 with "alveolitis," 22 without "alveolitis") and in 10 healthy controls. These were compared to clinical measures of lung disease and "alveolitis" in the SLS patients. RESULTS: SP-D levels were 300+/-214 ng/ml (mean+/-SD) in the SSc patients compared to 40+/-51 ng/ml in controls (p<0.0001). KL-6 levels were 1225+/-984 U/ml in the SSc patients and 333+/-294 U/ml in controls (p<0.0001). SSc patients with "alveolitis" had higher levels of both SP-D and KL-6 than those without "alveolitis." The level of SP-D was 353+/-219 ng/ml in patients with "alveolitis" and 161+/-143 ng/ml without "alveolitis" (p=0.0002). The level of KL 6 was 1458+/-1070 U/ml in patients with "alveolitis" and 640+/-487 U/ml without "alveolitis" (p=0.0001). Receiver operator characteristic curve analysis demonstrated high sensitivity and specificity of both SP-D and KL-6 for the determination of "alveolitis." KL-6 and SP-D were positively correlated with maximum fibrosis scores, but not with maximum ground-glass opacities, on HRCT. CONCLUSION: Serum levels of SP-D and KL-6 appear to be indicative of "alveolitis" in SSc patients as defined by the SLS, and are significantly higher than in SSc patients without "alveolitis." Serum SP-D and KL-6 may serve as noninvasive serological means of assessing interstitial lung disease in patients with SSc. PMID- 19286850 TI - The gap between practice and guidelines in the choice of first-line disease modifying antirheumatic drug in early rheumatoid arthritis: results from the ESPOIR cohort. AB - OBJECTIVE: To compare rheumatologists' prescription for first disease modifying antirheumatic drug (DMARD) in early rheumatoid arthritis (RA) in real-life settings with 2 clinical practice guidelines (CPG), the French Society of Rheumatology/STPR 2004 and EULAR/ESCISIT 2007, and thus assess the gap between practices and guidelines. Method. ESPOIR was a French multicenter cohort study of 813 patients with early arthritis between 2002 and 2005. "Definite" and "probable" RA were defined according to ACR criteria and the level of diagnostic certainty. The objectives were to assess conformity between the observed first line DMARD prescribed for those patients and the DMARD recommended in the guidelines; and to conduct a mail survey of patients' usual rheumatologists to investigate the reasons for their nonconformity with guidelines. RESULTS: In total 627 patients with definite or probable RA were identified. Conformity rates were 58% for STPR guidelines and 54% for EULAR guidelines. At 6 months, 83 (34%) patients with early RA did not receive any DMARD. Main determinants associated with conformity to guidelines were disease activity and presence of severity predictive factors. The main reason leading to a discrepancy between guidelines and daily practice appeared to be diagnostic uncertainty, i.e., the difficulty to reliably assess RA diagnosis as early as the first visits to the rheumatologist. CONCLUSION: There is a substantial gap between CPG and rheumatologists' daily practice concerning the first DMARD to prescribe in early RA. This is explained mainly by diagnostic uncertainty. More attention should be paid in future guidelines to the diagnostic difficulties of early RA. PMID- 19286851 TI - Implementation of Z-scores as an age- and sex-independent parameter for estimating joint space widths in rheumatoid arthritis. AB - OBJECTIVE: To compare normative data of joint space distances (JSD) with the JSD of patients with rheumatoid arthritis (RA) as measured by computer-aided joint space analysis (CAJSA) at the metacarpophalangeal (MCP) articulations, and to differentiate age- and sex-related alterations from the disease-related joint space narrowing. METHODS: In total, 256 healthy subjects and 248 patients with verified RA (following revised ACR criteria) underwent computerized semiautomated measurements of JSD (CAJSA, version 1.3.6) at the MCP articulation (JSD-MCP) based on digital radiographs. The Z-score, a comparative parameter that differentiates joint space alterations caused by RA-related cartilage destruction from age- and sex-related changes, was calculated. RESULTS: Our data showed a relationship between measured joint space widths (MCP total and MCP thumb to little finger) and age for healthy subjects and also the RA group. The RA group revealed an age-related joint space narrowing that was surpassed by the RA related narrowing of joint space widths classified by Sharp joint space narrowing score and resulting in smaller Z-scores for RA patients. CONCLUSION: The CAJSA technique seems to distinguish age-related JSD changes in healthy volunteers from RA-induced alterations. In addition the Z-score was also able to differentiate RA dependent narrowing of JSD. Calculation of the Z-scores based on sex- and age specific reference data may facilitate earlier identification of patients with RA, allowing initiation of a more optimal, individually adapted therapeutic strategy. PMID- 19286852 TI - Gout in the elderly--a population health study. AB - OBJECTIVE: To determine the incidence, healthcare utilization, and costs in older adults with gout. METHODS: A 5-year retrospective case-control study of patients with incident gout and matched controls was performed. Study variables were derived from health administrative data and included patient demographics, International Classification of Diseases diagnostic codes, and healthcare cost information. RESULTS: There were 4,071 cases and 16,281 controls, providing a 5 year incidence of gout of 4.4%. The mean (+/-SD) age (77+/-7.3 and 76+/-7.1 yrs) and the male:female ratio (1.0:1.04) were similar in both groups. Gout was diagnosed by family physicians (77%), nonrheumatology subspecialists (18%), general internists (4%), and rheumatologists (0.02%). Hospitalizations were significantly higher in cases (p<0.001) in the year of diagnosis. Patients with gout had an average of 28.1 physician visits per year compared to 20.6 for controls (p<0.0001). Drug utilization for the treatment (nonsteroidal antiinflammatory drugs, colchicine, corticosteroids) and prevention (allopurinol, probenecid, sulfinpyrazone) of gout was significantly higher (p<0.0001). The average healthcare cost differential was +$134 (Cdn) per month (p<0.001) and +$8,020 per case over 5 years. These costs were due to hospital utilization (64.4%), medications (23.1%), and physician visits (12.5%). CONCLUSION: Gout is associated with a high disease burden in older men and women. The cost is primarily attributable to hospitalization, probably due to the comorbidities associated with gout. As the majority of cases are managed by nonrheumatologists, it is important that guidelines for the diagnosis and treatment of gout are disseminated to and met by all physician groups. PMID- 19286853 TI - CENP-O, a protein localized at the centromere throughout the cell cycle, is a novel target antigen in systemic sclerosis. AB - OBJECTIVE: CENP-A, -B, and -C are major centromere components and the main targets of anticentromere antibodies (ACA). Many other proteins are also assembled around CENP-A nucleosomes in interphase nuclei to form the interphase centromere complex (ICEN). The CENP-O protein is a component of the ICEN that localizes at the centromere throughout the cell cycle. We investigated whether CENP-O is also targeted by sera from patients with systemic autoimmune diseases. METHODS: Sera from 114 patients with ACA and 142 patients without ACA were analyzed. Western blotting and an ELISA with bacterially expressed recombinant CENP-O protein were performed to screen for the presence of anti-CENP-O antibodies. In addition, anti-CENP-O antibody-positive sera were tested by Western blotting HeLa cell extracts to examine reactivity with the major centromere antigens. RESULTS: Four female patients with ACA had anti-CENP-O antibodies. There was no correlation of anti-CENP-O antibodies with specific clinical features or other serological features. However, one of the 4 patients, who showed a unique clinical course of scleroderma, had sera with markedly high reactivity to CENP-O. CONCLUSION: CENP-O protein is a novel centromere antigen that is recognized by a very minor population of ACA-positive patients with scleroderma. Because CENP-O is an ICEN component, ICEN may be a large antigenic structure in systemic autoimmunity. PMID- 19286854 TI - Osteoarthritis Measurement in Routine Rheumatology Outpatient Practice (OMIRROP) in Australia: a survey of practice style, instrument use, responder criteria, and state-attainment criteria. AB - OBJECTIVE: The purpose of the 2007 Osteoarthritis Measurement in Routine Rheumatology Outpatient Practice survey was to describe practice styles, instrument usage, and perceptions of responder criteria and state-attainment criteria in osteoarthritis (OA) management in routine clinical rheumatology practice. METHODS: A 16-item questionnaire (65 subcomponents) was developed, pretested, revised, formatted, and mailed to rheumatologists residing in Australia. Responses were obtained from 136 rheumatologists (response rate 58%). RESULTS: Approximately half the Australian respondents did not follow up their patients with hip and knee OA and two-thirds did not follow up their patients with hand OA. Health status measures (HSM) were infrequently used, even by those respondents who followed their patients with OA longitudinally, and the scores from those HSM that were used, were rarely if ever formally recorded. Respondents rated the following 6 requirements of a measure for use in clinical practice as very important: validity, reliability, responsiveness, simplicity, quick completion, and easy scoring. One-fifth to one-quarter of respondents indicated they did not know quantitatively what constituted a clinically important improvement, or a health state acceptable to patients with OA. The majority of the remainder selected values not closely aligned with published values in the peer review literature. CONCLUSION: While simply describing the health status of the patient is interesting, the more strategic applications are in benchmarking, and using the data to inform shared decision-making and therapeutic goal-setting. The OMIRROP survey suggests that further investigation of interpretation issues are essential, before evaluating the role of quantitative measurement in routine OA clinical practice. PMID- 19286856 TI - Do tumor necrosis factor inhibitors cause malformations in humans? PMID- 19286855 TI - Prevalence of hip symptoms and radiographic and symptomatic hip osteoarthritis in African Americans and Caucasians: the Johnston County Osteoarthritis Project. AB - OBJECTIVE: To report contemporary estimates of the prevalence of hip-related osteoarthritis (OA) outcomes in African Americans and Caucasians aged>or=45 years. METHODS: Weighted prevalence estimates and their corresponding 95% confidence intervals for hip symptoms, radiographic hip OA, symptomatic hip OA, and severe radiographic hip OA were calculated using SUDAAN for age, race, and sex subgroups among 3068 participants (33% African Americans, 38% men) in the baseline examination (1991-97) of The Johnston County Osteoarthritis Project, a population-based study of OA in North Carolina. Radiographic hip OA was defined as Kellgren-Lawrence radiographic grade>or=2, moderate/severe radiographic hip OA as grades 3 and 4, and symptomatic hip OA as hip symptoms in a hip with radiographic OA. RESULTS: Hip symptoms were present in 36%; 28% had radiographic hip OA; nearly 10% had symptomatic hip OA; and 2.5% had moderate/severe radiographic hip OA. Prevalence of all 4 outcomes was higher in older individuals; most outcomes were higher for women and African Americans. CONCLUSION: African Americans in this population do not have a lower prevalence of hip-related OA outcomes as previous studies suggested. Increasing public and health system awareness of the relatively high prevalence of these outcomes, which can be disabling, may help to decrease their effects and ultimately prevent them. PMID- 19286857 TI - Cardiovascular outcomes in systemic lupus erythematosus: big studies for big questions. PMID- 19286858 TI - The challenges of developing online learning. PMID- 19286859 TI - Optimizing limb position for measuring knee anatomical axis alignment from standing knee radiographs. PMID- 19286860 TI - Bisphosphonate associated osteonecrosis of the jaw. AB - In 2003, the first reports describing osteonecrosis of the jaw (ONJ) in patients receiving bisphosphonates (BP) were published. These cases occurred in patients with cancer receiving high-dose intravenous BP; however, 5% of the cases were in patients with osteoporosis receiving low-dose bisphosphonate therapy. We present the results of a systematic review of the incidence, risk factors, diagnosis, prevention, and treatment of BP associated ONJ. We conducted a comprehensive literature search for relevant studies on BP associated ONJ in oncology and osteoporosis patients published before February 2008.All selected relevant articles were sorted by area of focus. Data for each area were abstracted by 2 independent reviewers. The results showed that the diagnosis is made clinically. Prospective data evaluating the incidence and etiologic factors are very limited. In oncology patients receiving high-dose intravenous BP, ONJ appears to be dependent on the dose and duration of therapy, with an estimated incidence of 1% 12% at 36 months of exposure. In osteoporosis patients, it is rare, with an estimated incidence < 1 case per 100,000 person-years of exposure. The incidence of ONJ in the general population is not known. Currently, there is insufficient evidence to confirm a causal link between low-dose BP use in the osteoporosis patient population and ONJ. We concluded BP associated ONJ is associated with high-dose BP therapy primarily in the oncology patient population. Prevention and treatment strategies are currently based on expert opinion and focus on maintaining good oral hygiene and conservative surgical intervention. PMID- 19286861 TI - Catastrophic antiphospholipid syndrome in the central nervous system. PMID- 19286862 TI - Computed tomography angiography of type III Takayasu arteritis. PMID- 19286863 TI - Abdominal radiograph in systemic sclerosis. PMID- 19286864 TI - A forgotten cause of central nervous system vasculitis. PMID- 19286865 TI - GRAPPA at the European League Against Rheumatism (EULAR) 2008. PMID- 19286866 TI - A unique articular manifestation in a child with relapsing polychondritis. PMID- 19286867 TI - New onset of inflammatory polyarthritis in a patient taking adalimumab. PMID- 19286868 TI - A nice approach to TADs. PMID- 19286869 TI - British Orthodontic Society--Maurice Berman Prize 2007. AB - This paper describes the orthodontic treatment of a multidisciplinary case, which was awarded the Maurice Berman Prize in 2007 at the British Orthodontic Conference, Harrogate. PMID- 19286870 TI - How to ... take a wax bite for a Twin Block appliance. AB - The Twin Block appliance is the most commonly used functional appliance in the UK. It is very important to take a satisfactory wax bite, as it not only helps the technician to make an accurate appliance but it is essential for the successful use of this appliance. In this article, we suggest the appropriate clinical technique for taking an effective wax bite for the Twin Block appliance. PMID- 19286872 TI - Frictional resistance in plastic preadjusted brackets ligated with low-friction and conventional elastomeric ligatures. AB - OBJECTIVE: To compare frictional resistance of the plastic preadjusted brackets ligated with the low-friction ligatures with those of the conventional elastomeric ligatures. DESIGN: In vitro study. SETTING: Department of Orthodontics, Faculty of Dentistry, Kyushu University, Fukuoka, Japan. MATERIALS AND METHODS: The testing model consisted of four 0.022-inch plastic preadjusted brackets for the first premolar, the canine, the lateral incisor, and the central incisor. A superelastic nickel-titanium 0.014-inch wire and a stainless steel 0.019 x 0.025-inch wire were used for this test. The brackets were either aligned or out of line by 0.5, 1.0, 1.5 and 2.0 mm for the 0.014-inch wire and aligned for the 0.019 x 0.025-inch wire. The frictional forces in plastic preadjusted brackets with low-friction ligatures and conventional elastomeric ligatures were measured at a pulling speed of 0.1 mm/second. Welch t-tests were used to compare the mean differences of each testing measurement between the low-friction and the conventional elastomeric ligatures. RESULTS: In both use of the superelastic nickel-titanium 0.014-inch wire and the stainless steel 0.019 x 0.025-inch wire, the brackets with the low-friction ligatures showed significantly lower frictional forces than those of the conventional elastomeric ligatures in both aligned and all misaligned brackets (P < 0.0001). CONCLUSION: The study found the significantly lower frictional forces for the low-friction ligatures than those of the conventional elastomeric ligatures. PMID- 19286873 TI - Comparison of the Bolton Standards to longitudinal cephalograms superimposed on the occipital condyle (I-point). AB - OBJECTIVE: To compare traditional superimposition on sella turcica and the anterior cranial base (SACB) to superimposition referenced at the occipital condyle (I-point) for demonstrating craniofacial growth and development. MATERIALS AND METHODS: Tracings for ages 8, 10, 12, 15 and 18 were chosen from the Bolton Standards of Dentofacial Developmental Growth to compare superimposition with the traditional longitudinal reference at SACB (anterior curvature of sella turcica and anterior cranial base) to reference at I-point on the antero-inferior contour of the occipital condyles in norma lateralis. The serial tracings were superimposed using both the sagittal and postero-anterior (PA) tracings. Incremental growth of landmarks was measured in relation to Cartesian coordinates and compared between the superimposition methods. RESULTS: Sagittal and PA tracing superimpositions displayed an average 7 mm greater cephalad movement of landmarks, an average 2.4 mm greater ventral movement, and comparable transverse dimension with superimposition referenced at I-point as compared to the SACB reference. CONCLUSION: I-point superimposition demonstrates physiologic growth patterns concealed by traditional registration at sella turcica. The evolution of superimposition on SACB was based on convenience and reproducibility. Fundamental principles of bone development are consistent with the occipital condyles as a more biologic reference for relative craniofacial growth. Actual vertical growth is believed to be greater than displayed in this study, due to the cross-sectional and blended nature of the sample. PMID- 19286874 TI - From alginate impressions to digital virtual models: accuracy and reproducibility. AB - OBJECTIVE: To compare the accuracy and reproducibility of measurements performed on digital virtual models with those taken on plaster casts from models poured immediately after the impression was taken, the 'gold standard', and from plaster models poured following a 3-5 day shipping procedure of the alginate impression. DESIGN: Direct comparison of two measuring techniques. SETTING: The study was conducted at the Department of Orthodontics, School of Dentistry, University of Aarhus, Denmark in 2006/2007. PARTICIPANTS: Twelve randomly selected orthodontic graduate students with informed consent. METHODS: Three sets of alginate impressions were taken from the participants within 1 hour. Plaster models were poured immediately from two of the sets, while the third set was kept in transit in the mail for 3-5 days. Upon return a plaster model was poured as well. Finally digital models were made from the plaster models. A number of measurements were performed on the plaster casts with a digital calliper and on the corresponding digital models using the virtual measuring tool of the accompanying software. Afterwards these measurements were compared statistically. RESULTS: No statistical differences were found between the three sets of plaster models. The intra- and inter-observer variability are smaller for the measurements performed on the digital models. CONCLUSIONS: Sending alginate impressions by mail does not affect the quality and accuracy of plaster casts poured from them afterwards. Virtual measurements performed on digital models display less variability than the corresponding measurements performed with a calliper on the actual models. PMID- 19286875 TI - A systematic review of clinical trials of aligning archwires. AB - OBJECTIVES: This review aimed to identify the evidence for the efficacy of archwires used in the alignment stage of orthodontic treatment by undertaking a systematic review of the literature. DATA SOURCES: MEDLINE, the Cochrane Central Register of Controlled trials (CENTRAL), EMBASE, and the meta Register of Controlled Trials were searched up to July 2008. Reference lists of identified articles and relevant review articles were checked for further possible studies. REVIEW METHODS: Controlled clinical trials and randomised clinical trials that compared aligning archwires and reported objective measures of alignment were selected for inclusion. Validity and quality assessment were undertaken to identify studies with a low risk of bias. Details of the study methodology and the reported results were then abstracted. RESULTS: 100 studies were identified by the searches and 7 of these were identified as meeting the selection criteria. Four studies were deemed, after quality assessment, to have a low risk of bias and data was extracted from these. No two studies shared a common methodology or common reporting of outcome. Meta-analysis was therefore not possible. CONCLUSIONS: There is insufficient data in these studies to make clear recommendations regarding the most effective archwire for alignment. Recommendations on future study design have been made. PMID- 19286876 TI - Structured abstracts: do they improve citation retrieval from dental journals? AB - OBJECTIVES: To assess whether structured abstracts improved the sensitivity, precision and yield of retrieving clinical trials, using electronic searches, for example, MEDLINE, from dental journals. DESIGN: Retrospective, observational study. SAMPLE: Clinical trials, published in six dental journals. Three that adopted structured abstracts (BDJ, CPJ, JO) and three that remained unchanged (JDR, EJO, AJODO) between January 1995 and December 1998 (extended to December 2002 for the JO). INTERVENTION: Adoption of a structured abstract format. CONTROL: Continued use of a non-structured abstract format. METHOD: A combination of handsearching and the Cochrane Collaboration Oral Health Group's Trials Register and/or CENTRAL were used to identify randomised controlled trials (RCTs) and controlled clinical trials (CCTs) over the selected time period. MEDLINE was used to identify clinical trials in the selected journals over the same time period. RESULTS: There was no significant difference in the sensitivity or yield of clinical trial retrieval in journals with either abstract format over time. However, there was a significant increase in precision in journals that did not change their format (OR=4.96 (95% CI 1.18, 20.86) but not those that did. There was no significant difference in the sensitivity or yield of clinical trial retrieval either before or after the change in format or precision of retrieval before the change. However, in the later period, the precision of retrieval was significantly better in journals with unstructured abstracts compared to those with structured abstracts (OR=0.17 (95% CI 0.04, 0.7). CONCLUSION: The use of a structured abstract format does not improve the sensitivity, precision or yield of retrieval of clinical trials from MEDLINE. PMID- 19286877 TI - Orthognathic treatment: see how they feel? AB - As clinicians we are becoming increasingly careful in our pre-treatment screening processes and in acknowledging the importance of psychological assessment of potential orthognathic patients. However, this does not necessarily guarantee post-treatment satisfaction, even if the clinician thinks the clinical outcome is good. This paper provides the clinician with a schematic framework of those factors which may affect post-treatment outcomes. PMID- 19286879 TI - A new mutation in the hepcidin promoter impairs its BMP response and contributes to a severe phenotype in HFE related hemochromatosis. AB - Low levels of hepcidin are responsible for the development of iron overload in p.Cys282Tyr HFE related hemochromatosis. Every genetic factor lowering the hepcidin gene expression could contribute to a more severe phenotype in HFE hemochromatosis. Based on this hypothesis, we identified a heterozygous nc.-153 C>T mutation in the hepcidin gene promoter sequence in a patient homozygous for the p.Cys282Tyr HFE mutation who presented massive iron overload, resisting to well conducted iron depletive treatment. Our results demonstrate that the nc.-153 C>T mutation, located within a BMP-RE (Bone Morphogenetic Protein-Responsive Element): i) decreases the transcriptional activity of the hepcidin promoter, ii) alters its IL-6 (Interleukin-6) total responsiveness, and iii) prevents the binding of the SMAD protein complex (1/5/8 and 4) to the BPM-RE. In conclusion, our results suggest that a mutation in the BMP-RE of hepcidin promoter may impact on human iron metabolism. PMID- 19286880 TI - Autosomal dominant C1149R von Willebrand disease: phenotypic findings and their implications. AB - BACKGROUND: Mutation C1149R in the von Willebrand factor (VWF) gene has been thought to cause autosomal dominant severe type 1 von Willebrand disease (VWD). DESIGN AND METHODS: Eight patients from three unrelated families with this mutation were included in the present study who had distinct VWF abnormalities, not described in earlier studies. RESULTS: The patients showed notably low levels of VWF antigen (VWF:Ag), VWF ristocetin cofactor activity (VWF:RCo), VWF collagen binding (VWF:CB), and a reduced ristocetin-induced platelet aggregation (RIPA). VWF:RCo/VWF:Ag and VWF:CB/VWF:Ag ratios were lower than 0.7. At basal conditions, all the VWF multimers were decreased in plasma, with a clearly lower relative proportion of the high molecular weight VWF multimers (HMWM). In high-resolution agarose gels, a large decrease in the relative proportions of the satellite bands was seen. The patients had a brief good response to desmopressin (DDAVP) administration, but the released VWF half-life was shorter than normal, indicating an accelerated clearance of their VWF. Platelet VWF was abnormal. CONCLUSIONS: We conclude from the results obtained in these patients for plasma phenotypic data that this mutation should be classified as a VWD type 2A (IIE). DDAVP therapy may be somewhat helpful for this mutation, at least for mild to moderate bleeding. These data provide evidence that for VWD classification factors other than basal VWF, such as DDAVP response and platelet VWF, should be considered. PMID- 19286881 TI - Improved treatment feasibility in children with hemophilia using arteriovenous fistulae: the results after seven years of follow-up. AB - BACKGROUND: An easy and stable venous access is essential in hemophilic children who receive regular prophylaxis or immune tolerance induction treatment. Central venous access devices improve treatment feasibility, but their use is complicated by infection and/or thrombosis. Arteriovenous fistula (AVF) has been evaluated as an alternative to central venous access devices in hemophilic children since 1999. DESIGN AND METHODS: This study provides results obtained in a large series after seven years of follow-up. RESULTS: From 1999 to 2008, 43 procedures were performed in 38 children (median age: 2.7 years). Thirty-five AVFs (81%) achieved maturation after a median of 58 days and were used for a median of five years (range: 0.4-8.5). A brachial artery caliber larger than 1.2 mm was associated with successful maturation (p<0.05). Complications with some impact on arteriovenous fistula use or duration were observed in 14/43 procedures (32%) and in 13/38 children (34%). Age at arteriovenous fistula creation was younger in children who lost arteriovenous fistula patency (p<0.05) and aneurysms were more frequent in children who were on daily treatment regimen and thus had a greater cumulative number of arteriovenous fistula accesses (p<0.05). At the end of the follow-up period, 22 AVFs were still in use and 9 had been surgically dismantled. Arteriovenous fistula use allowed long-term prophylaxis (up to 8.5 years) in 11 children and the completion of immune tolerance induction without interruptions in 18 children. CONCLUSIONS: This study confirms the feasibility of arteriovenous fistula with an acceptable rate of complications and suggests that its use is particularly favorable in children with inhibitors in whom it should be considered as first-choice venous access. PMID- 19286882 TI - A novel (epsilongammadeltabeta)(o)-thalassemia deletion associated with an alpha globin gene triplication leading to a severe transfusion dependent fetal thalassemic syndrome. PMID- 19286883 TI - F9 Malmo, factor IX and deep vein thrombosis. AB - BACKGROUND: We recently reported the association between the Malmo sequence variant in F9 (rs6048) and deep vein thrombosis. DESIGN AND METHODS: We aimed to study whether the association between F9 Malmo and deep vein thrombosis is explained by linkage disequilibrium with nearby single-nucleotide polymorphisms, and whether the association is explained biologically by F9 Malmo affecting factor IX antigen levels or activation of factor IX. We investigated the association of F9 Malmo and 28 nearby single-nucleotide polymorphisms with deep vein thrombosis in men from two case-control studies, LETS (n=380) and MEGA (n=1,469). We assessed the association of F9 Malmo with factor IX antigen level in male control subjects from LETS (n=191) and two subsets of MEGA (n=823 and n=484) and the association with endogenous thrombin potential in LETS control men. We studied the association between F9 Malmo and factor IX activation peptide in 1,199 healthy middle-aged men from the NPHS-II cohort. RESULTS: In the combined LETS and MEGA studies, the odds ratio (95% confidence interval) for the G allele of F9 Malmo, compared with the A allele, was 0.80 (0.69-0.93). One single-nucleotide polymorphism in F9, rs422187, was strongly linked to F9 Malmo (r(2)=0.94) and was similarly associated with deep vein thrombosis. No other single-nucleotide polymorphism or haplotype tested was more strongly associated. Factor IX antigen level, factor IX activation peptide levels and endogenous thrombin potential did not differ between F9 Malmo genotypes. CONCLUSIONS: The F9 Malmo sequence variant was the most strongly associated with deep vein thrombosis among common single-nucleotide polymorphisms in the region. However, the biological mechanism by which F9 Malmo affects risk remains unknown. PMID- 19286885 TI - Platelet receptors and signaling in the dynamics of thrombus formation. AB - Hemostasis and pathological thrombus formation are dynamic processes that require a co-ordinated series of events involving platelet membrane receptors, bidirectional intracellular signals, and release of platelet proteins and inflammatory substances. This review aims to summarize current knowledge in the key steps in the dynamics of thrombus formation, with special emphasis on the crucial participation of platelet receptors and signaling in this process. Initial tethering and firm adhesion of platelets to the exposed subendothelium is mediated by glycoprotein (GP) Ib/IX/V complex and collagen receptors, GP VI and alpha(2)beta(1) integrin, in the platelet surface, and by VWF and fibrillar collagen in the vascular site. Interactions between these elements are largely influenced by flow and trigger signaling events that reinforce adhesion and promote platelet activation. Thereafter, soluble agonists, ADP, thrombin, TxA(2), produced/released at the site of vascular injury act in autocrine and paracrine mode to amplify platelet activation and to recruit circulating platelets to the developing thrombus. Specific interactions of these agonists with their G-protein coupled receptors generate inside-out signaling leading to conformational activation of integrins, in particular alpha(IIb)beta(3), increasing their ligand affinity. Binding of alpha(IIb)beta(3) to its ligands, mainly fibrinogen, supports processes such as clot retraction and platelet aggregation. Stabilization of thrombi is supported by the late wave of signaling events promoted by close contact between aggregated platelets. The best known contact dependent signaling is outside-in signaling through alphaIb beta(3), but new ones are being clarified such as those mediated by interaction of Eph receptors with ephrins, or by Sema 4D and Gas-6 binding to their receptors. Finally, newly identified mechanisms appear to control thrombus growth, including back-shifting of activated integrins and actuation of compensatory molecules such as ESAM or PECAM-1. The expanding knowledge of thrombotic disease is expected to translate into the development of new drugs to help management and prevention of thrombosis. PMID- 19286886 TI - Elevated risk of chronic lymphocytic leukemia and other indolent non-Hodgkin's lymphomas among relatives of patients with chronic lymphocytic leukemia. AB - BACKGROUND: Previous studies have shown increased familial risk for chronic lymphocytic leukemia. In the most comprehensive study to date, we evaluated risk of chronic lymphocytic leukemia and lymphoproliferative disorders among first degree relatives of chronic lymphocytic leukemia cases compared to first-degree relatives of controls. DESIGN AND METHODS: Population-based registry data from Sweden were used to evaluate outcomes in 26,947 first-degree relatives of 9,717 chronic lymphocytic leukemia patients (diagnosed 1958-2004) compared with 107,223 first-degree relatives of 38,159 matched controls. Using a marginal survival model, we calculated relative risks (RR) and 95% confidence intervals as measures of familial aggregation. RESULTS: Compared to relatives of controls, relatives of chronic lymphocytic leukemia patients had an increased risk for chronic lymphocytic leukemia (RR=8.5, 6.1-11.7) and other non-Hodgkin's lymphomas (NHLs) (RR=1.9, 1.5-2.3). Evaluating NHL subtypes, we found a striking excess of indolent B-cell NHL, specifically lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia and hairy cell leukemia. No excesses of aggressive B-cell or T cell lymphomas were found. There was no statistical excess of Hodgkin's lymphoma, multiple myeloma, or the precursor condition, monoclonal gammopathy of undetermined significance, among chronic lymphocytic leukemia relatives. CONCLUSIONS: These familial aggregations are striking and provide novel clues to research designed to uncover early pathogenetic mechanisms in chronic lymphocytic leukemia including studies to identify germ line susceptibility genes. However, clinicians should counsel their chronic lymphocytic leukemia patients emphasizing that because the baseline population risks are low, the absolute risk for a first degree relative to develop chronic lymphocytic leukemia or another indolent lymphoma is low. At this time, an increased medical surveillance of first-degree relatives of chronic lymphocytic leukemia patients has no role outside research studies. PMID- 19286887 TI - Exploring psychophysiological markers of vulnerability to somatic illnesses in females. AB - OBJECTIVE: To examine the association between biological stress regulation and somatic complaints in young girls prior to the onset of clear psychopathology such as somatization disorder. METHODS: Salivary cortisol, heart rate variability (HRV), and negative mood were assessed in 48 12-year-old girls in response to the Trier Social Stress Test for Children (TSST-C). Parent and child report on the Children's Somatization Inventory was used to identify girls with high and low somatic complaints. RESULTS: Girls with high levels of somatic complaints had significantly higher initial levels of cortisol, which decreased over time, and showed a trend for a more limited HRV in response to the TSST-C than girls with low levels of somatic complaints. CONCLUSIONS: High levels of cortisol and possibly low HRV among girls with somatic complaints may interfere with flexibility in responding to typical psychosocial stressors, which may increase vulnerability to the onset of somatic illnesses in females. PMID- 19286888 TI - Severity of sickle cell disease in Yemeni children. PMID- 19286889 TI - Did Darwin read Mendel? PMID- 19286890 TI - Epilepsy genetics: clinical beginnings and social consequences. AB - The approach to epilepsy care has transformed in the last 30 years, with more and better anti-epileptic medications, improved cerebral imaging and increased surgical options. Alongside this, developments in neuroscience and molecular genetics have furthered the understanding of epileptogenesis. Future developments in pharmacogenomics hold the promise of antiepileptic drugs matched to specific genotypes. Despite this rapid progress, one-third of epilepsy patients remain refractory to medication, with their seizures impacting upon day-to-day activity, social well-being, independence, economic output and quality of life. International genome collaborations, such as HapMap and the Welcome Trust Case Control Consortium single nucleotide polymorphism (SNP) mapping project have identified common genetic variations in diseases of major public health importance. Such genetic signposts should help to identify at-risk populations with a view to producing more effective pharmaceutical treatments. Neurological disorders, despite comprising one-fifth of UK acute medical hospital admissions, are surprisingly under-represented in these projects. Epilepsy is the commonest serious neurological disorder worldwide. Although physically, psychologically, socially and financially disabling, it rarely receives deserved attention from physicians, scientists and governmental bodies. As outlined in this article, research into epilepsy genetics presents unique challenges. These help to explain why the identification of its complex genetic traits has lagged well behind other disciplines, particularly the efforts made in neuropsychiatric disorders. Clinical beginnings must underpin any genetic understanding in epilepsy. Success in identifying genetic traits in other disorders does not make the automatic case for genome-wide screening in epilepsy, but such is a desired goal. The essential clinical approach of accurately phenotyping, diagnosing and interpreting the dynamic nature of epilepsy remains fundamental to harvesting its potential translational outcomes. PMID- 19286891 TI - Myocardial infarction after percutaneous coronary intervention: a meta-analysis of troponin elevation applying the new universal definition. AB - AIM: Elevation of Troponin after scheduled percutaneous coronary intervention (PCI) is a recognized consequence. We sought to evaluate the prognostic significance and impact of the newly published definition of PCI-related myocardial infarction (MI) according to which any troponin elevation >3 times the upper reference limit identify a peri-procedural MI. METHODS: Search of BioMedCentral, CENTRAL, mRCT and PubMed (updated May 2008). Outcomes of interest were: MACE [the composite of all cause death, MI, repeat target vessel PCI (re PCI) and coronary artery bypass grafting (CABG)]; single end points were also assessed. RESULTS: Fifteen studies have been included totalling 7578 patients. Troponin elevation occurred in 28.7% of the procedures. The incidence of PCI related MI according to the new definition was 14.5%. During the hospitalization, any level of raised troponin was associated with an increased risk of MACE [OR 11.29 (3.00-42.48), Number needed to harm (NNH) 5], death [OR 7.16 (1.95-26.27), NNH = 100], MI [OR 30.85 (6.05-157.38), NNH = 4] and re-PCI [OR 4.13 (1.23 13.88), NNH = 50]. Patients with PCI-related MI had an increased risk of death [OR 17.25 (2.71-109.96), NNH = 100] and re-PCI [OR 10.86 (3.2-36.94), NNH = 25]. At follow up of 18 months any troponin elevation was associated with an increased risk of MACE [OR 1.48 (1.12-1.96), NNH = 20], death [OR 2.19 (1.59-3.00), NNH = 50], MI [OR 3.29 (2.71-6.31), NNH = 33] and re-PCI [OR 1.47 (1.06-2.03), NNH = 25]. In patients with PCI-related MI the risk of MACE was further increased: OR 2.25 (1.26-4.00), NNH = 3. An increase of the troponin level below the cut-off was not associated with MACE. CONCLUSION: A diagnosis of MI according to the new guidelines applies to 15% of patients undergoing PCI and these patients are at high risk of further adverse events both during the hospital stay and at 18 months. PMID- 19286892 TI - Refinements in husbandry, care and common procedures for non-human primates: Ninth report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement. AB - Preface Whenever animals are used in research, minimizing pain and distress and promoting good welfare should be as important an objective as achieving the experimental results. This is important for humanitarian reasons, for good science, for economic reasons and in order to satisfy the broad legal principles in international legislation. It is possible to refine both husbandry and procedures to minimize suffering and improve welfare in a number of ways, and this can be greatly facilitated by ensuring that up-to-date information is readily available. The need to provide such information led the British Veterinary Association Animal Welfare Foundation (BVAAWF), the Fund for the Replacement of Animals in Medical Experiments (FRAME), the Royal Society for the Prevention of Cruelty to Animals (RSPCA) and the Universities Federation for Animal Welfare (UFAW) to establish a Joint Working Group on Refinement (JWGR) in the UK. The chair is Professor David Morton and the secretariat is provided by the RSPCA. This report is the ninth in the JWGR series. The RSPCA is opposed to the use of animals in experiments that cause pain, suffering, distress or lasting harm and together with FRAME has particular concerns about the continued use of non-human primates. The replacement of primate experiments is a primary goal for the RSPCA and FRAME. However, both organizations share with others in the Working Group, the common aim of replacing primate experiments wherever possible, reducing suffering and improving welfare while primate use continues. The reports of the refinement workshops are intended to help achieve these aims. This report produced by the British Veterinary Association Animal Welfare Foundation (BVAAWF)/Fund for the Replacement of Animals in Medical Experiments (FRAME)/Royal Society for the Prevention of Cruelty to Animals (RSPCA)/Universities Federation for Animal Welfare (UFAW) Joint Working Group on Refinement (JWGR) sets out practical guidance on refining the husbandry and care of non-human primates (hereinafter primates) and on minimizing the adverse effects of some common procedures. It provides a valuable resource to help understand the physical, social and behavioural characteristics and needs of individual primates, and is intended to develop and complement the existing literature and legislative guidelines. Topics covered include refinements in housing, husbandry and common procedures such as restraint, identification and sampling, with comprehensive advice on issues such as primate communication, assessing and facilitating primate wellbeing, establishing and maintaining social groups, environmental and nutritional enrichment and animal passports. The most commonly used species are the key focus of this resource, but its information and recommendations are generally applicable to other species, provided that relevant individual species characteristics are taken into account. PMID- 19286893 TI - Computer-tailored dietary behaviour change interventions: a systematic review. AB - Improving dietary behaviours such as increasing fruit and vegetable consumption and reducing saturated fat intake are important in the promotion of better health. Computer tailoring has shown promise as a strategy to promote such behaviours. A narrative systematic review was conducted to describe the available evidence on 'second'-generation computer-tailored primary prevention interventions for dietary behaviour change and to determine their effectiveness and key characteristics of success. Systematic literature searches were conducted through five databases: Medline, Embase, PsycINFO, CINAHL and All EBM Reviews and by examining the reference lists of relevant articles to identify studies published in English from January 1996 to 2008. Randomized controlled trials or quasi-experimental designs with pre-test and post-test behavioural outcome data were included. A total of 13 articles were reviewed, describing the evaluation of 12 interventions, seven of which found significant positive effects of the computer-tailored interventions for dietary behaviour outcomes, one also for weight reduction outcomes. Although the evidence of short-term efficacy for computer-tailored dietary behaviour change interventions is fairly strong, the uncertainty lies in whether the reported effects are generalizable and sustained long term. Further research is required to address these limitations of the evidence. PMID- 19286894 TI - Planned, motivated and habitual hygiene behaviour: an eleven country review. AB - Handwashing with soap (HWWS) may be one of the most cost-effective means of preventing infection in developing countries. However, HWWS is rare in these settings. We reviewed the results of formative research studies from 11 countries so as to understand the planned, motivated and habitual factors involved in HWWS. On average, only 17% of child caretakers HWWS after the toilet. Handwash 'habits' were generally not inculcated at an early age. Key 'motivations' for handwashing were disgust, nurture, comfort and affiliation. Fear of disease generally did not motivate handwashing, except transiently in the case of epidemics such as cholera. 'Plans' involving handwashing included to improve family health and to teach children good manners. Environmental barriers were few as soap was available in almost every household, as was water. Because much handwashing is habitual, self-report of the factors determining it is unreliable. Candidate strategies for promoting HWWS include creating social norms, highlighting disgust of dirty hands and teaching children HWWS as good manners. Dividing the factors that determine health-related behaviour into planned, motivated and habitual categories provides a simple, but comprehensive conceptual model. The habitual aspects of many health-relevant behaviours require further study. PMID- 19286895 TI - The influence of tobacco marketing on adolescent smoking intentions via normative beliefs. AB - Using cross-sectional data from three waves of the Youth Tobacco Policy Study, which examines the impact of the UK's Tobacco Advertising and Promotion Act (TAPA) on adolescent smoking behaviour, we examined normative pathways between tobacco marketing awareness and smoking intentions. The sample comprised 1121 adolescents in Wave 2 (pre-ban), 1123 in Wave 3 (mid-ban) and 1159 in Wave 4 (post-ban). Structural equation modelling was used to assess the direct effect of tobacco advertising and promotion on intentions at each wave, and also the indirect effect, mediated through normative influences. Pre-ban, higher levels of awareness of advertising and promotion were independently associated with higher levels of perceived sibling approval which, in turn, was positively related to intentions. Independent paths from perceived prevalence and benefits fully mediated the effects of advertising and promotion awareness on intentions mid- and post-ban. Advertising awareness indirectly affected intentions via the interaction between perceived prevalence and benefits pre-ban, whereas the indirect effect on intentions of advertising and promotion awareness was mediated by the interaction of perceived prevalence and benefits mid-ban. Our findings indicate that policy measures such as the TAPA can significantly reduce adolescents' smoking intentions by signifying smoking to be less normative and socially unacceptable. PMID- 19286896 TI - Effects of HIV-1 infection in vitro on transendothelial migration by monocytes and monocyte-derived macrophages. AB - Monocytes constitutively migrate from the bloodstream across the vascular endothelium for systemic immune surveillance and maintenance of macrophage populations. They also perform reverse transendothelial migration (TEM) across the endothelium, which is required for entry of tissue monocytes/macrophages into the lymphatics or back into the bloodstream. We have modeled these processes previously using HUVEC monolayers grown on three-dimensional collagen matrices. The aim of the present study was to determine whether HIV-1 infection of monocytes/macrophages in vitro affects TEM. Purified primary human monocytes and monocyte-derived macrophages (MDM) expressed important TEM proteins such as CD62L, CD18, PECAM-1, CCR2, and CCR8. Purified monocytes underwent efficient forward and reverse TEM across HUVEC, and this function was maintained by MDM after up to 15 days of culture. Monocytes exposed to HIV-1 for 2 days had unaltered forward or reverse TEM. However, HIV-1 infection of MDM for 7 days decreased reverse TEM by an average of 66.5% compared with mock-infected MDM (n=9 independent donors; P=0.004), without affecting forward TEM. Decreased reverse TEM by HIV-infected MDM required viral RT and was not a result of alterations in surface expression of CCR8 or p-glycoprotein or a general impairment in mobility, as assessed by migration toward fMLP. This study indicates that HIV-1 infection of macrophages reduces their capacity to emigrate from the subendothelial extracellular matrix in vitro, which could result in defective cell-mediated immune responses to infections and promote establishment of viral reservoirs of HIV in tissue macrophages in vivo. PMID- 19286897 TI - Pendulous heart with tamponade. PMID- 19286899 TI - Moderate patient-prosthesis mismatch can impact on mortality after aortic valve replacement. PMID- 19286900 TI - Coronary calcium scoring: the first step in case of suspected significant coronary atherosclerosis. PMID- 19286901 TI - Stroke and migraine: a cardiologist's headache. PMID- 19286902 TI - Role of adrenergic receptors in human coronary vasomotion. PMID- 19286903 TI - Dyslipidaemia, statins and rheumatoid arthritis. PMID- 19286904 TI - Guidelines for initiation of anti-tumour necrosis factor therapy in rheumatoid arthritis: similarities and differences across Europe. PMID- 19286905 TI - Interpreting lipid levels in the context of high-grade inflammatory states with a focus on rheumatoid arthritis: a challenge to conventional cardiovascular risk actions. AB - In severe untreated rheumatoid arthritis (RA), reductions in high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol and total cholesterol have been noted; this is in line with findings in other pathologies/conditions associated with inflammation or infection, such as sepsis, cancer, trauma or the postoperative period. Although the precise mechanisms remain to be established, cytokine-induced activation of the reticuloendothelial system is potentially critical to such changes. Consequently, dampening of inflammation in severe RA-as occurs with several biologics-may lead to increases, not only in high-density lipoprotein-cholesterol, but also with other lipid moieties, including total and low-density lipoprotein-cholesterol and, perhaps, triglycerides. This concept is consistent with findings following antitumour necrosis factor treatment and interleukin-6 receptor inhibition in patients with RA. At the same time, it is increasingly apparent that potent dampening of inflammation, however achieved, broadly reduces the risk of cardiovascular disease in RA. Therefore, changes in lipid profiles, particularly increases in cholesterol and triglycerides that occur with treatments for severe inflammation, may not represent increased cardiovascular risk as in the usual understanding of lipid-level elevations in individuals without significant inflammation. Rather, changes in lipid levels, in part or largely, may represent a predictable response to attenuation of inflammation. These observations are increasingly important clinically and should aid in the understanding and interpretation of lipid changes under inflammatory conditions, as well as in the context of potent anti inflammatory interventions. PMID- 19286906 TI - The effect of joint position on Doppler flow in finger synovitis. PMID- 19286907 TI - Decreased plasma IL22 levels, but not increased IL17 and IL23 levels, correlate with disease activity in patients with systemic lupus erythematosus. PMID- 19286908 TI - Severe bronchospasm associated with rituximab for refractory Churg-Strauss syndrome. PMID- 19286909 TI - Rituximab therapy for refractory systemic-onset juvenile idiopathic arthritis. PMID- 19286910 TI - Arthroscopic repair for chronic anterior shoulder instability: a comparative study between patients with Bankart lesions and patients with combined Bankart and superior labral anterior posterior lesions. AB - BACKGROUND: Although labrum lesions in patients with chronic anterior shoulder instability may not only involve detachment of the anteroinferior labrum but a lesion of the superior glenoid labrum as well, no studies have compared the clinical outcome between patients with a lesion of the anteroinferior labrum and patients with a combined lesion of the anterior and superior labrum after arthroscopic shoulder stabilization. HYPOTHESIS: Arthroscopic repair of a combined lesion of the anterior and superior labrum may have inferior clinical outcome to repair of an anterior lesion only in patients with anterior shoulder instability. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Sixty three patients operated on for anterior shoulder instability between April 2002 and June 2006 were included in this study. Patients with bone deficiency were excluded. Fixation of the detached labrum was performed using suture anchors. Thirty-eight patients had a lesion of the anterior labrum (group A), and 25 had a combined lesion of the anterior and superior labrum (group B). Patients were evaluated after a 2-year minimum follow-up with Constant and Rowe scores. Failure was defined as a redislocation or a subluxation episode. RESULTS: Patients in group B experienced a significantly higher number of dislocations preoperatively (P < .05). However, there was no difference between the 2 groups regarding the failure rate postoperatively. One patient from each group had a failed result. A mean loss of 5 degrees and 8 degrees of external rotation at 90 degrees of abduction was noted in patients in groups A and B, respectively (P = .113). The Constant score was 94 in group A and 93 in group B (P = .435). The Rowe score was 91 in group A and 90 in group B (P = .338). CONCLUSION: There are no differences in shoulder stability and function in patients with anterior shoulder instability and a lesion of the anteroinferior labrum and patients with an extended lesion of the anterior and superior labrum after arthroscopic shoulder stabilization. PMID- 19286911 TI - Outcomes of autologous chondrocyte implantation in a diverse patient population. AB - BACKGROUND: Autologous chondrocyte implantation is indicated as a second-line treatment of large, irregularly shaped chondral defects after failure of first line surgical intervention. This study examines the clinical results of a patient cohort undergoing autologous chondrocyte implantation and elucidates factors associated with subjective improvement after implantation. HYPOTHESIS: Autologous chondrocyte implantation will result in long-term functional and symptomatic improvement. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The cohort included 137 subjects (140 knees) who underwent autologous chondrocyte implantation of the knee. Mean defect size per patient was 5.2 +/- 3.5 cm(2) (range, 0.8-26.6 cm(2)). Patients averaged 30.3 +/- 9.1 years of age (range, 13.9 49.9 years) and were followed for 4.3 +/- 1.8 years (range, 2.0-9.7 years). Outcomes were assessed via clinical assessment and established outcome scales, including the Lysholm scale, International Knee Documentation Committee scale, and Short Form-12. RESULTS: A significant improvement after surgery was observed in all outcome assessments including the Lysholm (41 to 69; P < .001) and International Knee Documentation Committee (34 to 64; P < .001) scales. Subjectively, 75% of patients indicated they were completely or mostly satisfied with the outcome and 83% would have the procedure again. Preoperatively, 32% of patients had a Tegner score of 6 or greater, compared with 82% before injury and 65% at most recent follow-up. Multivariate analysis identified age (P < .021) and receiving workers' compensation (P < .018) as independent predictors of follow-up Lysholm score. Twenty-one patients (16%) required debridement of the autologous chondrocyte implantation site secondary to persistent symptoms, whereas 9 knees (6.4%) clinically failed and underwent a revision procedure. CONCLUSION: Autologous chondrocyte implantation is a viable treatment option for chondral defects of the knee, resulting in durable functional and symptomatic improvement. Age and workers' compensation status are independent predictors of outcome. PMID- 19286912 TI - Treatment of recalcitrant iliotibial band friction syndrome with open iliotibial band bursectomy: indications, technique, and clinical outcomes. AB - BACKGROUND: Iliotibial band friction syndrome (ITBFS) is an overuse injury causing lateral knee pain. There is evidence that the pathological lesion is in fact an inflamed bursa underlying the iliotibial band (ITB) rather than an inflamed ITB itself. HYPOTHESIS: Resection of the bursa underlying the ITB in ITBFS patients will relieve their pain and allow them to return to their preinjury activity level. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: We describe the technique of ITB bursectomy and report a minimal 20 month follow-up of patients who had ITB bursectomies performed by a single surgeon. The patients completed a survey detailing their preoperative and postoperative symptoms and activities. RESULTS: The senior author performed 12 consecutive cases of ITB bursectomies (12 patients). One was excluded from the study (previous microfracture). The average age at surgery was 32 years (standard deviation, 5; range, 24-41). There were 7 men and 4 women. Postoperatively, patients were able to return to their preinjury Tegner activity levels, and the visual analog pain scores decreased by an average of 6 points (P < .001). Six patients were completely satisfied with the surgical outcome, 3 were mostly satisfied, 2 were somewhat satisfied, and none were dissatisfied. Nine of 11 patients said that knowing what they know now, they would have the surgery performed again for the same problem. CONCLUSION: Iliotibial band bursectomy successfully reduces knee pain in patients with ITBFS and allows them to return to their preinjury level of activity. The great majority of patients were satisfied with the results of the procedure. PMID- 19286913 TI - Injury patterns and injury rates in the circus arts: an analysis of 5 years of data from Cirque du Soleil. AB - BACKGROUND: Human circus arts are gaining increasing popularity as a physical activity with more than 500 companies and 200 schools. The only injury data that currently exist are a few case reports and 1 survey. HYPOTHESIS: To describe injury patterns and injury rates among Cirque du Soleil artists between 2002 and 2006. STUDY DESIGN: Descriptive epidemiology study. METHODS: The authors defined an injury as any work-related condition recorded in an electronic injury database that required a visit to the show therapist. Analyses for treatments, missed performances, and injury rates (per 1000 artist performances) were based on a subset of data that contained appropriate denominator (exposure) information (began in 2004). RESULTS: There were 1376 artists who sustained a total of the 18 336 show- or training-related injuries. The pattern of injuries was generally similar across sex and performance versus training. Most injuries were minor. Of the 6701 injuries with exposure data, 80% required < or =7 treatments and resulted in < or =1 completely missed performance. The overall show injury rate was 9.7 (95% confidence interval, 9.4-10.0; for context, published National Collegiate Athletic Association women's gymnastics rate was 15.2 injuries per 1000 athlete-exposures). The rate for injuries resulting in more than 15 missed performances for acrobats (highest risk group) was 0.74 (95% confidence interval, 0.65-0.83), which is much lower than the corresponding estimated National Collegiate Athletic Association women's gymnastics rate. CONCLUSION: Most injuries in circus performers are minor, and rates of more serious injuries are lower than for many National Collegiate Athletic Association sports. PMID- 19286914 TI - Knee medial compartment contact pressure increases with release of the type I anterior intermeniscal ligament. AB - BACKGROUND: The anterior intermeniscal ligament of the knee is at risk during knee arthroscopy, anterior cruciate ligament reconstruction, and tibial nail insertion. HYPOTHESIS: Release of the anterior intermeniscal ligament, in knees with type I ligaments, will result in altered contact pressures in the medial compartment. STUDY DESIGN: Controlled laboratory study. METHODS: Five fresh frozen human cadaveric knees with intact type I anterior intermeniscal ligaments were chosen for testing in a modified MTS machine from 0 degrees to 60 degrees of flexion under 2 conditions: (1) intact and (2) after sharp sectioning of the anterior intermeniscal ligament. Measurements were made using inframeniscal contact pressure sensors covering the medial compartment. Poststudy analysis was done in 10 degrees increments between 0 degrees and 60 degrees of flexion, looking at peak contact pressure and the amount of contact area seeing pressure. RESULTS: Sectioning of the anterior intermeniscal ligament caused a statistically significant increase in the peak pressure at 20 degrees , 30 degrees , 40 degrees , and 50 degrees of knee flexion. The largest change occurred at 40 degrees of knee flexion, when the peak pressure increased by 27.5% (3.68 MPa to 4.69 MPa). Contact area decreased, although this difference was not statistically significant. CONCLUSION: Release of the anterior intermeniscal ligament results in increased peak contact pressures in the medial compartment of the knee. CLINICAL RELEVANCE: Care should be taken to avoid sacrifice of this ligament during surgery. PMID- 19286915 TI - Characterization of leaf apoplastic peroxidases and metabolites in Vigna unguiculata in response to toxic manganese supply and silicon. AB - Previous work suggested that the apoplastic phenol composition and its interaction with apoplastic class III peroxidases (PODs) are decisive in the development or avoidance of manganese (Mn) toxicity in cowpea (Vigna unguiculata L.). This study characterizes apoplastic PODs with particular emphasis on the activities of specific isoenzymes and their modulation by phenols in the Mn sensitive cowpea cultivar TVu 91 as affected by Mn and silicon (Si) supply. Si reduced Mn-induced toxicity symptoms without affecting the Mn uptake. Blue Native PAGE combined with Nano-LC-MS/MS allowed identification of a range of POD isoenzymes in the apoplastic washing fluid (AWF). In Si-treated plants Mn mediated induction of POD activity was delayed. Four POD isoenzymes eluted from the BN gels catalysed both H(2)O(2)-consuming and H(2)O(2)-producing activity with pH optima at 6.5 and 5.5, respectively. Four phenols enhanced NADH peroxidase activity of these isoenzymes in the presence of Mn(2+) (p coumaric=vanillic>>benzoic>ferulic acid). p-Coumaric acid-enhanced NADH peroxidase activity was inhibited by ferulic acid (50%) and five other phenols (50-90%). An independent component analysis (ICA) of the total and apoplastic GC MS-based metabolome profile showed that Mn, Si supply, and the AWF fraction (AWF(H(2)O), AWF(NaCl)) significantly changed the metabolite composition. Extracting non-polar metabolites from the AWF allowed the identification of phenols. Predominantly NADH-peroxidase activity-inhibiting ferulic acid appeared to be down-regulated in Mn-sensitive (+Mn, -Si) and up-regulated in Mn-tolerant (+Si) leaf tissue. The results presented here support the previously hypothesized role of apoplastic NADH-peroxidase and its activity-modulating phenols in Mn toxicity and Si-enhanced Mn tolerance. PMID- 19286916 TI - Characterization of two cotton cDNAs encoding trans-2-enoyl-CoA reductase reveals a putative novel NADPH-binding motif. AB - Very long chain fatty acids are important components of plant lipids, suberins, and cuticular waxes. Trans-2-enoyl-CoA reductase (ECR) catalyses the fourth reaction of fatty acid elongation, which is NADPH dependent. In the present study, the expression of two cotton ECR (GhECR) genes revealed by quantitative RT PCR analysis was up-regulated during cotton fibre elongation. GhECR1 and 2 each contain open reading frames of 933 bp in length, both encoding proteins consisting of 310 amino acid residues. GhECRs show 32% identity to Saccharomyces cerevisiae Tsc13p at the deduced amino acid level, and the GhECR genes were able to restore the viability of the S. cerevisiae haploid tsc13-deletion strain. A putative non-classical NADPH-binding site in GhECR was predicted by an empirical approach. Site-directed mutagenesis in combination with gas chromatography-mass spectrometry analysis suggests that G(5X)IPXG presents a putative novel NADPH binding motif of the plant ECR family. The data suggest that both GhECR genes encode functional enzymes harbouring non-classical NADPH-binding sites at their C termini, and are involved in fatty acid elongation during cotton fibre development. PMID- 19286917 TI - HHP1 is involved in osmotic stress sensitivity in Arabidopsis. AB - HHP1 (heptahelical protein 1), a protein with a predicted seven transmembrane domain structure homologous to adiponectin receptors (AdipoRs) and membrane progestin receptors (mPRs), has been characterized. Expression of HHP1 was increased in response to abscisic acid (ABA) and salt/osmotic stress as shown by quantitative real-time PCR and HHP1 promoter-controlled GUS activity. The HHP1 T DNA insertion mutant (hhp1-1) showed a higher sensitivity to ABA and osmotic stress than the wild-type (WT), as revealed by the germination rate and post germination growth rate. The induced expression of stress-responsive genes (RD29A, RD29B, ADH1, KIN1, COR15A, and COR47) was more sensitive to exogenous ABA and osmotic stress in hhp1-1 than in the WT. The hypersensitivity in the hhp1-1 mutant was reversed in the complementation mutant of HHP1 expressing the HHP1 gene. The data suggest that the mutation of HHP1 renders plants hypersensitive to ABA and osmotic stress and HHP1 might be a negative regulator in ABA and osmotic signalling. PMID- 19286918 TI - Plant phenology: a critical controller of soil resource acquisition. AB - Plant phenology, the timing of plant growth and development, is changing in response to global climate change. Changing temperature, soil moisture, nitrogen availability, light, and elevated CO(2) are all likely to affect plant phenology. Alteration of plant phenology by global climate change may alter the ability of plants to acquire soil resources (water and nutrients) by altering the timing and duration of the deployment of roots and leaves, which drive resource acquisition. The potential importance of phenologically-driven changes in soil resource acquisition for plant fitness and productivity have received little attention. General hypotheses are proposed for how plant acquisition of soil resources may be affected by the alteration of phenology. It is expected that the acquisition of mobile resources will be approximately proportional to total transpiration. Alteration of phenology that increases total transpiration should increase, while changes in phenology that reduce transpiration should decrease the acquisition of mobile resources. The acquisition of immobile resources will be approximately proportional to root length duration, thus changes in phenology that increase growth duration should increase the acquisition of immobile resources and vice versa. For both groups of resources, longer growing seasons would tend to increase resource acquisition, and shorter growing seasons would tend to decrease resource acquisition. In the case of resources that exhibit seasonal variability in availability, the synchrony of resource availability and acquisition capacity is important, and subject to disturbance by the alteration of phenology. PMID- 19286919 TI - Influence of leaf dry mass per area, CO2, and irradiance on mesophyll conductance in sclerophylls. AB - Leaf photosynthesis (A) is limited by mesophyll conductance (g(m)), which is influenced by both leaf structure and the environment. Previous studies have indicated that the upper bound for g(m) declines as leaf dry mass per area (LMA, an indicator of leaf structure) increases, extrapolating to zero at a LMA of about 240 g m(-2). No data exist on g(m) and its response to the environment for species with LMA values higher than 220 g m(-2). In this study, laboratory measurements of leaf gas exchange and in vivo chlorophyll a fluorescence were used concurrently to derive estimates of g(m) in seven species of the Australian sclerophyllous genus Banksia covering a wide range of LMA (130-480 g m(-2)). Irradiance and CO(2) were varied during those measurements to gauge the extent of environmental effects on g(m). A significant decrease of g(m) with increasing LMA was found. g(m) declined by 35-60% in response to increasing atmospheric CO(2) concentrations at high irradiance, with a more variable response (0-60%) observed at low irradiance, where g(m) was, on average, 22% lower than at high irradiance at ambient CO(2) concentrations. Despite considerable variation in A and LMA between the Banksia species, the CO(2) concentrations in the intercellular air spaces (C(i), 262+/-5 micromol mol(-1)) and in the chloroplasts (C(c), 127+/-4 micromol mol(-1)) were remarkably stable. PMID- 19286920 TI - Is habitual alcohol drinking associated with reduced electrophoretic DNA migration in peripheral blood leukocytes from ALDH2-deficient male Japanese? AB - Alcohol drinking-derived acetaldehyde is believed to cross-link DNA and induce sister chromatid exchanges in peripheral blood lymphocytes. However, little population data are available to illustrate effects of alcohol-derived acetaldehyde on DNA migration as assayed by the comet assay in peripheral lymphocytes. In the present study, we investigated lifestyle behaviours, including alcohol consumption, in 150 Japanese males by questionnaire, determined their aldehyde dehydrogenase 2 (ALDH2) family genotypes by polymerase chain reaction and measured the DNA migration in peripheral blood leukocytes by the alkaline comet assay. The results showed that habitual alcohol drinking is significantly negatively associated with DNA migration in peripheral blood leukocytes (r = -0.321, P = 0.005) of ALDH2-deficient, but not of ALDH2 proficient genotypes (r = 0.048, P = 0.683). The amount of pure alcohol consumed per time by the subjects showed a similar phenomenon (r = -0.257, P = 0.025 for the ALDH2-deficient, but r = -0.061, P = 0.606 for the ALDH2-proficient genotype). Further stepwise multiple regression analysis showed that alcohol drinking frequency was a significant predictor of DNA migration for subjects with ALDH2-deficient genotype, but not for subjects with ALDH2-proficient genotype. In summary, the present result suggests that frequent alcohol drinking is significantly associated with a reduced electrophoretic DNA migration in peripheral blood leukocytes from ALDH2-deficient male Japanese subjects. PMID- 19286921 TI - The protein synthesis inhibitor anisomycin induces macrophage apoptosis in rabbit atherosclerotic plaques through p38 mitogen-activated protein kinase. AB - Because macrophages play a major role in atherosclerotic plaque destabilization, selective removal of macrophages represents a promising approach to stabilize plaques. We showed recently that the protein synthesis inhibitor cycloheximide, in contrast to puromycin, selectively depleted macrophages in rabbit atherosclerotic plaques without affecting smooth muscle cells (SMCs). The mechanism of action of these two translation inhibitors is dissimilar and could account for the differential effects on SMC viability. It is not known whether selective depletion of macrophages is confined to cycloheximide or whether it can also be achieved with translation inhibitors that have a similar mechanism of action. Therefore, in the present study, we investigated the effect of anisomycin, a translation inhibitor with a mechanism of action similar to cycloheximide, on macrophage and SMC viability. In vitro, anisomycin induced apoptosis of macrophages in a concentration-dependent manner, whereas SMCs were only affected at higher concentrations. In vivo, anisomycin selectively decreased the macrophage content of rabbit atherosclerotic plaques through apoptosis. The p38 mitogen-activated protein kinase (MAPK) inhibitor SB202190 [4-(4 fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)-1H-imidazole] prevented anisomycin-induced macrophage death, without affecting SMC viability. SB202190 decreased anisomycin-induced p38 MAPK phosphorylation, did not alter c-Jun NH(2) terminal kinase (JNK) phosphorylation, and increased extracellular signal regulated kinase (ERK) 1/2 phosphorylation. The latter effect was abolished by the mitogen-activated protein kinase kinase 1/2 inhibitor U0126 [1,4-diamino-2,3 dicyano-1,4-bis(2-aminophynyltio)butadiene ethanolate], although the prevention of anisomycin-induced macrophage death by SB202190 remained unchanged. The JNK phosphorylation inhibitor SP600125 did not affect anisomycin-induced macrophage or SMC death. In conclusion, anisomycin selectively decreased the macrophage content in rabbit atherosclerotic plaques, indicating that this effect is not confined to cycloheximide. p38 MAPK, but not ERK1/2 or JNK, plays a major role in anisomycin-induced macrophage death. PMID- 19286922 TI - Therapeutic letters as relationally responsive practice. AB - Therapeutic letters have become a practice adopted by many clinicians as an adjunct to therapeutic conversations. This article presents selected findings from a larger study that examined both the letter-writing practices of nine clinicians as well as the experiences of seven adult clients who received a therapeutic letter or letters during the course of individual or family therapy. A novel aspect of this study is that data from clients were gathered in the form of letters-eight letters written by the clients to the researcher about their experience of receiving a therapeutic letter or letters from their clinician. Thematic analysis guided the analysis of the data. Findings are placed within the context of relationally responsive (or relationally engaged) practice. An invitation is extended to conceptualize letters not as monologic documents but as a means of dialogically relating to clients and to the "goings-on" of therapeutic conversations. PMID- 19286924 TI - A severe deficiency of coagulation factor VIIa results in attenuation of the asthmatic response in mice. AB - Eosinophil counts in the bronchoalveolar lavage fluid of wild-type (WT) mice increased after ovalbumin (OVA) challenge, a response that was diminished in comparably challenged low-expressing coagulation factor VII (FVII(tTA/tTA)) mice. Levels of T helper type 2 (Th2) cytokines, IL-4, IL-5, and IL-13, and eosinophil attracting chemokines, eotaxin and RANTES, were also lower in the OVA-challenged FVII(tTA/tTA) mice. Eosinophils purified from low-FVII mice underwent apoptosis at a faster rate compared with WT eosinophils, and eosinophil migration in response to eotaxin was reduced in eosinophils obtained from FVII(tTA/tTA) mice. Airway hyperresponsiveness and mucous layer thickness were reduced in OVA-treated FVII(tTA/tTA) mice, and addition of exogenous coagulation factor X (FX) enhanced mucin production in human epithelial NCI-H292 cells. Correspondingly, incubation of FX with NCI-H292 cells resulted in activated (a) FX production, suggesting that the components required for FX activation were present on NCI-H292 cells. These results demonstrate that FVIIa functions in the asthmatic response to an allergen by stimulating lung eosinophilia, airway hyperresponsiveness, and mucin production, this latter effect through its ability to activate FX in conjunction with tissue factor. PMID- 19286925 TI - Secretory group V phospholipase A2 regulates acute lung injury and neutrophilic inflammation caused by LPS in mice. AB - We investigated the regulatory role of 14-kDa secretory group V phospholipase A(2) (gVPLA(2)) in the development of acute lung injury (ALI) and neutrophilic inflammation (NI) caused by intratracheal administration of LPS. Experiments were conducted in gVPLA(2) knockout (pla2g5(-/-)) mice, which lack the gene, and gVPLA(2) wild-type littermate control (pla2g5(+/+)) mice. Indices of pulmonary injury were evaluated 24 h after intratracheal administration of LPS. Expression of gVPLA(2) in microsections of airways and mRNA content in lung homogenates were increased substantially in pla2g5(+/+) mice after LPS-administered compared with saline-treated pla2g5(+/+) mice. By contrast, expression of gVPLA(2) was neither localized in LPS- nor saline-treated pla2g5(-/-) mice. LPS also caused 1) reduced transthoracic static compliance, 2) lung edema, 3) neutrophilic infiltration, and 4) increased neutrophil myeloperoxidase activity in pla2g5(+/+) mice. These events were attenuated in pla2g5(-/-) mice exposed to LPS or in pla2g5(+/+) mice receiving MCL-3G1, a neutralizing MAb directed against gVPLA(2), before LPS administration. Our data demonstrate that gVPLA(2) is an inducible protein in pla2g5(+/+) mice but not in pla2g5(-/-) mice within 24 h after LPS treatment. Specific inhibition of gVPLA(2) with MCL-3G1 or gene-targeted mice lacking gVPLA(2) blocks ALI and attenuates NI caused by LPS. PMID- 19286926 TI - alpha,beta-Unsaturated aldehydes contained in cigarette smoke elicit IL-8 release in pulmonary cells through mitogen-activated protein kinases. AB - Cigarette smoking is the major risk factor for chronic obstructive pulmonary disease (COPD), a syndrome characterized by pulmonary neutrophil infiltration, chronic inflammation, and progressive tissue destruction. We examined here the acute effect of aqueous cigarette smoke extract (CSE) and of two alpha,beta unsaturated aldehydes (acrolein and crotonaldehyde) contained in CSE in cultured normal human lung fibroblasts and small airway epithelial cells. By examining a panel of 19 cytokines and chemokines, we found that IL-8 release was elevated by CSE as well as by acrolein, whereas other inflammatory mediators were mostly unaffected. CSE-evoked IL-8 release was mimicked by acrolein and crotonaldehyde at concentrations (3-60 microM each) found in CSE and fully prevented by 1 mM alpha,beta-unsaturated aldehydes scavengers N-acetylcysteine (NAC) or sodium 2 mercaptoethanesulfonate. Neither the saturated aldehyde acetaldehyde nor H(2)O(2) evoked IL-8 release. In addition, CSE or crotonaldehyde upregulated the release of IL-8 from alveolar macrophages from both COPD patients and healthy nonsmokers, indicating that this is a response common to cells involved in lung inflammation. CSE-evoked IL-8 release was accompanied by increased phosphorylation of p38 MAPK and ERK1/2. CSE-evoked p38 and ERK1/2 phosphorylation was mimicked by acrolein and inhibited by NAC. IL-8 release elicited by both acrolein and CSE was blocked by pharmacological inhibition of p38 and ERK1/2 phosphorylation. In summary, our data show that alpha,beta-unsaturated aldehydes-evoked phosphorylation of p38 and ERK1/2 underlies IL-8 release elicited by CSE, thus shedding light on the mechanisms through which cigarette smoke can initiate inflammation in the lung. PMID- 19286927 TI - Ceramide-dependent PP2A regulation of TNFalpha-induced IL-8 production in respiratory epithelial cells. AB - IL-8 is a key mediator in the pathophysiology of acute lung injury. TNFalpha stimulates IL-8 production in respiratory epithelial cells by activating both the NF-kappaB and MAP kinase pathways. The precise mechanism by which these pathways are downregulated to terminate IL-8 production remains unclear. We studied the regulatory role of the serine/threonine phosphatase, PP2A, on the signaling pathways involved in IL-8 production from respiratory epithelial cells. Inhibition of PP2A using okadaic acid or gene knockdown using siRNA resulted in an augmentation of TNFalpha-induced IL-8 production. We also found that PP2A inhibition resulted in prolonged activation of JNK, p38, and ERK resulting in both increased transcriptional activation of the IL-8 promoter and posttranscriptional stabilization of IL-8 mRNA. Because TNFalpha had been shown to activate ceramide accumulation, and separate studies had linked ceramide with activation of PP2A, we hypothesized the pathway of TNFalpha-inducing ceramide to activate PP2A comprised an endogenous regulatory pathway. Inhibition of the immediate sphingomyelinase-dependent pathway as well as the de novo synthesis pathway of ceramide production reduced serine/threonine phosphatase activity and augmented IL-8 production. These data suggest that ceramide plays a role in activating PP2A to terminate ongoing IL-8 production. In summary, our data suggest that in respiratory epithelium, TNFalpha induces ceramide accumulation, resulting in subsequent activation of PP2A, which targets those kinases responsible for transcriptional activation of IL-8. PMID- 19286928 TI - Identification of putative endothelial progenitor cells (CD34+CD133+Flk-1+) in endarterectomized tissue of patients with chronic thromboembolic pulmonary hypertension. AB - Chronic thromboembolic pulmonary hypertension (CTEPH) is characterized by a fibrotic thrombus persisting and obliterating the lumen of pulmonary arteries; its pathogenesis remains poorly defined. This study investigates a potential contribution for progenitor cell types in the development of vascular obliteration and remodeling in CTEPH patients. Endarterectomized tissue from patients undergoing pulmonary thromboendarterectomy was collected and examined for the structure and cellular composition. Our data show an organized fibrin network structure in unresolved thromboemboli and intimal remodeling in vascular wall tissues, characterized by smooth muscle alpha-actin (SM-alphaA)-positive cell proliferation in proximal regions (adjacent to thromboemboli) and neoangiogenesis/recanalization in distal regions (downstream from thromboemboli). Cells that are positively stained with CD34 and fetal liver kinase-1 (Flk-1) (CD34(+)Flk-1(+)) were identified in both the proximal and distal vascular tissues; a subpopulation of CD34(+)Flk-1(+)CD133(+) cells were further identified by immunostaining. Triple-positive cells are indicative of a population of putative endothelial progenitor cells or potential colony-forming units of endothelial cells. In addition, inflammatory cells (CD45(+)) and collagen secreting cells (procollagen-1(+)) were detected in the proximal vascular wall. Some of the CD34(+) cells in CTEPH cells isolated from proximal regions were also positive for SM-alphaA. Our data indicate that putative progenitor cell types are present in the neointima of occluded vessels of CTEPH patients. It is possible that the microenvironment provided by thromboemboli may promote these putative progenitor cells to differentiate and enhance intimal remodeling. PMID- 19286929 TI - Cigarette smoke-induced emphysema in A/J mice is associated with pulmonary oxidative stress, apoptosis of lung cells, and global alterations in gene expression. AB - Cigarette smoking is the major risk factor for developing chronic obstructive pulmonary disease, the fourth leading cause of deaths in the United States. Despite recent advances, the molecular mechanisms involved in the initiation and progression of this disease remain elusive. We used Affymetrix Gene Chip arrays to determine the temporal alterations in global gene expression during the progression of pulmonary emphysema in A/J mice. Chronic cigarette smoke (CS) exposure caused pulmonary emphysema in A/J mice, which was associated with pronounced bronchoalveolar inflammation, enhanced oxidative stress, and increased apoptosis of alveolar septal cells. Microarray analysis revealed the upregulation of 1,190, 715, 260, and 246 genes and the downregulation of 1,840, 730, 442, and 236 genes in the lungs of mice exposed to CS for 5 h, 8 days, and 1.5 and 6 mo, respectively. Most of the genes belong to the functional categories of phase I genes, Nrf2-regulated antioxidant and phase II genes, phase III detoxification genes, and others including immune/inflammatory response genes. Induction of the genes encoding multiple phase I enzymes was markedly higher in the emphysematous lungs, whereas reduced expression of various cytoprotective genes constituting ubiquitin-proteasome complex, cell survival pathways, solute carriers and transporters, transcription factors, and Nrf2-regulated antioxidant and phase II responsive genes was noted. Our data indicate that the progression of CS-induced emphysema is associated with a steady decline in the expression of various genes involved in multiple pathways in the lungs of A/J mice. Many of the genes discovered in this study could rationally play an important role in the susceptibility to CS-induced emphysema. PMID- 19286930 TI - Gene association studies in acute lung injury: replication and future direction. PMID- 19286931 TI - Functionally important role for arginase 1 in the airway hyperresponsiveness of asthma. AB - l-Arginine metabolism by the arginase and nitric oxide (NO) synthase (NOS) families of enzymes is important in NO production, and imbalances between these pathways contribute to airway hyperresponsiveness (AHR) in asthma. To investigate the role of arginase isozymes (ARG1 and ARG2) in AHR, we determined the protein expression of ARG1, ARG2, the NOS isozymes, and other proteins involved in l arginine metabolism in lung tissues from asthma patients and in acute (3-wk) and chronic (12-wk) murine models of ovalbumin-induced airway inflammation. Expression of ARG1 was increased in human asthma, whereas ARG2, NOS isoforms, and the other l-arginine-related proteins (i.e., cationic amino acid transporters 1 and 2, agmatinase, and ornithine decarboxylase) were unchanged. In the acute murine model of allergic airway inflammation, augmentation of ARG1 expression was similarly the most dramatic change in protein expression. However, ARG2, NOS1, NOS2, and agmatinase were also increased, whereas NOS3 expression was decreased. Arginase inhibition in vivo with nebulized S-(2-boronoethyl)-l-cysteine attenuated the methacholine responsiveness of the central airways in mice from the acute model. Further investigations in the chronic murine model revealed an expression profile that more closely paralleled the human asthma samples: only ARG1 expression was significantly increased. Interestingly, in the chronic mouse model, which generates a remodeling phenotype, arginase inhibition attenuated methacholine responsiveness of the central and peripheral airways. The similarity in arginase expression between human asthma and the chronic model and the attenuation of AHR after in vivo treatment with an arginase inhibitor suggest the potential for therapeutic modification of arginase activity in asthma. PMID- 19286932 TI - Is greater self-neglect severity associated with lower levels of physical function? AB - OBJECTIVE: This study examined the association between severity of self-neglect and physical function in a population of community-dwelling older adults. METHOD: Participants were older adults (N = 1,094) reported to the Chicago Department on Aging for suspected self-neglect from 1993 to 2005, who also participated in the Chicago Health Aging Project. The primary outcome of physical function was assessed using physical performance tests. Secondary outcomes were assessed using the Katz, Nagi, and Rosow-Breslau scales. Multiple regression models were used to assess these associations. RESULTS: After adjusting for confounders, higher self neglect severity scores were associated with lower physical performance testing (coefficient = -.062, p = .001). Greater self-neglect severity was also correlated with the reported number of impairments on the Katz, Nagi, and Rosow Breslau scales (coefficients = .024, .024, and .016, respectively, p = .001). DISCUSSION: Higher self-neglect severity is associated with lower levels of physical function among older adults. PMID- 19286933 TI - A binding resolution. PMID- 19286934 TI - Reevaluation of abscisic acid-binding assays shows that G-Protein-Coupled Receptor2 does not bind abscisic Acid. PMID- 19286935 TI - The Arabidopsis RING finger E3 ligase RHA2a is a novel positive regulator of abscisic acid signaling during seed germination and early seedling development. AB - The phytohormone abscisic acid (ABA) is well known for its regulatory roles in integrating environmental constraints with the developmental programs of plants. Here, we characterize the biological function of the Arabidopsis (Arabidopsis thaliana) RING-H2 protein RHA2a in ABA signaling. The rha2a mutant is less sensitive to ABA than the wild type during seed germination and early seedling development, whereas transgenic plants overexpressing RHA2a are hypersensitive, indicating that RHA2a positively regulates ABA-mediated control of seed germination and early seedling development. Double mutant analyses of rha2a with several known ABA-insensitive mutants suggest that the action of RHA2a in ABA signaling is independent of that of the transcription factors ABI3, ABI4, and ABI5. We provide evidence showing that RHA2a also positively regulates plant responses to salt and osmotic stresses during seed germination and early seedling development. RHA2a is a functional E3 ubiquitin ligase, and its conserved RING domain is likely important for the biological function of RHA2a in ABA signaling. Together, these results suggest that the E3 ligase RHA2a is an important regulator of ABA signaling during seed germination and early seedling development. PMID- 19286936 TI - The Arabidopsis callose synthase gene GSL8 is required for cytokinesis and cell patterning. AB - Cytokinesis is the division of the cytoplasm and its separation into two daughter cells. Cell plate growth and cytokinesis appear to require callose, but direct functional evidence is still lacking. To determine the role of callose and its synthesis during cytokinesis, we identified and characterized mutants in many members of the GLUCAN SYNTHASE-LIKE (GSL; or CALLOSE SYNTHASE) gene family in Arabidopsis (Arabidopsis thaliana). Most gsl mutants (gsl1-gsl7, gsl9, gsl11, and gsl12) exhibited roughly normal seedling growth and development. However, mutations in GSL8, which were previously reported to be gametophytic lethal, were found to produce seedlings with pleiotropic defects during embryogenesis and early vegetative growth. We found cell wall stubs, two nuclei in one cell, and other defects in cell division in homozygous gsl8 insertional alleles. In addition, gsl8 mutants and inducible RNA interference lines of GSL8 showed reduced callose deposition at cell plates and/or new cell walls. Together, these data show that the GSL8 gene encodes a putative callose synthase required for cytokinesis and seedling maturation. In addition, gsl8 mutants disrupt cellular and tissue-level patterning, as shown by the presence of clusters of stomata in direct contact and by islands of excessive cell proliferation in the developing epidermis. Thus, GSL8 is required for patterning as well as cytokinesis during Arabidopsis development. PMID- 19286937 TI - Suppression of a phospholipase D gene, OsPLDbeta1, activates defense responses and increases disease resistance in rice. AB - Phospholipase D (PLD) plays an important role in plants, including responses to abiotic as well as biotic stresses. A survey of the rice (Oryza sativa) genome database indicated the presence of 17 PLD genes in the genome, among which OsPLDalpha1, OsPLDalpha5, and OsPLDbeta1 were highly expressed in most tissues studied. To examine the physiological function of PLD in rice, we made knockdown plants for each PLD isoform by introducing gene-specific RNA interference constructs. One of them, OsPLDbeta1-knockdown plants, showed the accumulation of reactive oxygen species in the absence of pathogen infection. Reverse transcription-polymerase chain reaction and DNA microarray analyses revealed that the knockdown of OsPLDbeta1 resulted in the up-/down-regulation of more than 1,400 genes, including the induction of defense-related genes such as pathogenesis-related protein genes and WRKY/ERF family transcription factor genes. Hypersensitive response-like cell death and phytoalexin production were also observed at a later phase of growth in the OsPLDbeta1-knockdown plants. These results indicated that the OsPLDbeta1-knockdown plants spontaneously activated the defense responses in the absence of pathogen infection. Furthermore, the OsPLDbeta1-knockdown plants exhibited increased resistance to the infection of major pathogens of rice, Pyricularia grisea and Xanthomonas oryzae pv oryzae. These results suggested that OsPLDbeta1 functions as a negative regulator of defense responses and disease resistance in rice. PMID- 19286938 TI - Myocardial oxygenation and functional recovery in infarct rat hearts transplanted with mesenchymal stem cells. AB - Stem cell therapy for myocardial tissue repair is limited by the poor survival of transplanted cells, possibly because of inadequate supply of oxygen and nutrients. The purpose of this study was to assess the oxygenation level and functional recovery after allogenic transplantation of mesenchymal stem cells (MSC) in a rat model of myocardial infarction (MI). Myocardial oxygen tension (Po(2)) was measured by electron paramagnetic resonance oximetry using an implantable oxygen-sensing spin probe (OxySpin). MSCs incubated with OxySpins showed substantial uptake of the probe without affecting its oxygen sensitivity or calibration. The cells internalized with OxySpins were able to differentiate into osteogenic, adipogenic, cardiomyocyte, and endothelial cell lineages. The labeled cells tested positive for CD44 and CD29 markers and negative for the hematopoietic markers CD14 and CD45. For the in vivo studies, MI was induced in rats by permanently ligating the left anterior descending coronary artery. MSCs with OxySpins were transplanted in the infarct region of hearts. A significant increase in Po(2) was observed in the MSC group compared with the untreated MI group (18.1 +/- 2.6 vs. 13.0 +/- 1.8 mmHg, n = 4, P < 0.05) at 4 wk after transplantation. Echocardiography showed a significant improvement in ejection fraction and fraction shortening, which inversely correlated with the magnitude of fibrosis in the treated hearts. The cell-transplanted hearts also showed an increase in vascular endothelial growth factor level and capillary density in the infarct region. The study established our ability to measure and correlate changes in myocardial tissue oxygenation with cardiac function in infarcted rat hearts treated with MSCs. PMID- 19286939 TI - Voltage-independent calcium channels mediate lipopolysaccharide-induced hyporeactivity to endothelin-1 in the rat aorta. AB - The roles of intracellular calcium concentration ([Ca(2+)](i)) and Ca(2+) sensitization in lipopolysaccharide (LPS)-induced vascular smooth muscle (VSM) hyporesponsiveness are incompletely understood. To investigate these roles, contraction responses to endothelin-1 (ET-1) and 80 mM KCl; relaxation responses to nifedipine; the expression levels of mRNAs of ET-1 and its receptors (ET(A) or ET(B)); the expression levels of protein kinase C (PKC) and phosphorylation of Rho kinase (ROKalpha), CPI-17, and myosin phosphatase target subunit-1 (MYPT1); and changes in aortic VSM cell [Ca(2+)](i) were measured in LPS-treated aortic rings from male Wistar rats (250-300 g). LPS (10 mug/ml, 20 h) decreased contraction induced by ET-1 (0.3-100 nM) or 80 mM KCl. LPS-induced hypocontractility was not observed in the absence of external Ca(2+), but LPS treated aorta remained hypocontractile on subsequent stepwise restoration of extracellular Ca(2+) (0.01-10 mM). Vascular relaxation to nifedipine; mRNA expression levels of ET-1, ET(A), or ET(B); protein expression levels of PKC; and phosphorylation levels of ROKalpha, CPI-17, and MYPT1 were not affected by LPS. In isolated aortic VSM cells, ET-1 caused a transient initial increase in [Ca(2+)](i), followed by a maintained tonic increase in [Ca(2+)](i), which was decreased by LPS pretreatment and was dependent on external Ca(2+). Subsequent restoration of extracellular Ca(2+) increased [Ca(2+)](i), but this increase was lower in the LPS-treated group. This difference in response to extracellular Ca(2+) addition was not affected by diltiazem, but was abolished by SKF-96365. Therefore, LPS induces hyporeactivity to ET-1 in rat aorta that depends on external Ca(2+) influx through non-voltage-operated Ca(2+) channels, but not on ET-1 receptor expression or Ca(2+) sensitization. PMID- 19286940 TI - Regulation of nitrite transport in red blood cells by hemoglobin oxygen fractional saturation. AB - Allosteric regulation of nitrite reduction by deoxyhemoglobin has been proposed to mediate nitric oxide (NO) formation during hypoxia. Nitrite is predominantly an anion at physiological pH, raising questions about the mechanism by which it enters the red blood cell (RBC) and whether this is regulated and coupled to deoxyhemoglobin-mediated reduction. We tested the hypothesis that nitrite transport by RBCs is regulated by fractional saturation. Using human RBCs, nitrite consumption was faster at lower fractional saturations, consistent with faster reactions with deoxyheme. A membrane-based regulation was suggested by slower nitrite consumption with intact versus lysed RBCs. Interestingly, upon nitrite addition, intracellular nitrite concentrations attained a steady state that, despite increased rates of consumption, did not change with decreasing oxygen tensions, suggesting a deoxygenation-sensitive step that either increases nitrite import or decreases the rate of nitrite export. A role for anion exchanger (AE)-1 in the control of nitrite export was suggested by increased intracellular nitrite concentrations in RBCs treated with DIDS. Moreover, deoxygenation decreased steady-state levels of intracellular nitrite in AE-1 inhibited RBCs. Based on these data, we propose a model in which deoxyhemoglobin binding to AE-1 inhibits nitrite export under low oxygen tensions allowing for the coupling between deoxygenation and nitrite reduction to NO along the arterial to-venous gradient. PMID- 19286941 TI - Thyroid hormone analog 3,5-diiodothyropropionic acid promotes healthy vasculature in the adult myocardium independent of thyroid effects on cardiac function. AB - Patients with hypothyroidism are at a higher risk for coronary vascular disease. Patients with diabetes and related vascular complications also have an increased incidence of low thyroid function. While thyroid hormones (THs) may be key regulators of a healthy vasculature, potential undesirable side effects hinder their use in the treatment of vascular disorders. TH analogs such as 3,5 diiodothyropropionic acid (DITPA) may provide a safer treatment option. However, the relative potency of DITPA on vascular growth, cardiac function, and metabolism is poorly understood. We hypothesized that the vascular growth promoting effects of DITPA can be obtained with a minimum effect on cardiac function. Thyroidectomized Sprague-Dawley rats were given slow-release pellets with either thyroxine (T4, 2.7 or 5.2 mg) or DITPA (80 mg) for 6 wk and were compared with placebo. Heart mass, body mass, body temperature, serum THs, cardiac function (echocardiograms and hemodynamics), and myocardial arteriolar density were determined. Hypothyroidism led to reductions in cardiac function, heart mass, body temperature, and myocardial arterioles. High-dose T4 prevented arteriolar loss and the development of hypothyroidism. Low-dose T4 partially prevented the reduction in cardiac function but had minimal effects on arteriolar loss. In contrast, DITPA treatment prevented myocardial arteriolar loss but not the progression of hypothyroid-induced changes in cardiac function. The results suggested that DITPA can promote a healthy vasculature independently from its thyroid-related metabolic effects. Drugs in this class may provide new therapeutic options for patients with vascular disease. PMID- 19286942 TI - Paraplegia increased cardiac NGF content, sympathetic tonus, and the susceptibility to ischemia-induced ventricular tachycardia in conscious rats. AB - Midthoracic spinal cord injury is associated with ventricular arrhythmias that are mediated, in part, by enhanced cardiac sympathetic activity. Furthermore, it is well known that sympathetic neurons have a lifelong requirement for nerve growth factor (NGF). NGF is a neurotrophin that supports the survival and differentiation of sympathetic neurons and enhances target innervation. Therefore, we tested the hypothesis that paraplegia is associated with an increased cardiac NGF content, sympathetic tonus, and susceptibility to ischemia induced ventricular tachyarrhythmias. Intact and paraplegic (6-9 wk posttransection, T(5) spinal cord transection) rats were instrumented with a radiotelemetry device for recording arterial pressure, temperature, and ECG, and a snare was placed around the left main coronary artery. Following recovery, the susceptibility to ventricular arrhythmias (coronary artery occlusion) was determined in intact and paraplegic rats. In additional groups of matched intact and paraplegic rats, cardiac nerve growth factor content (ELISA) and cardiac sympathetic tonus were determined. Paraplegia, compared with intact, increased cardiac nerve growth factor content (2,146 +/- 286 vs. 180 +/- 36 pg/ml, P < 0.05) and cardiac sympathetic tonus (154 +/- 4 vs. 68 +/- 4 beats/min, P < 0.05) and decreased the ventricular arrhythmia threshold (3.6 +/- 0.2 vs. 4.9 +/- 0.2 min, P < 0.05). Thus altered autonomic behavior increases the susceptibility to ventricular arrhythmias in paraplegic rats. PMID- 19286943 TI - Air pollution and cardiac remodeling: a role for RhoA/Rho-kinase. AB - Exposure to ambient air pollution has been associated with increases in blood pressure. We have previously demonstrated activation of the Rho/Rho kinase pathway in experimental hypertension in rats. In this investigation, we evaluated the effects of particulate matter of < 2.5 microm (PM(2.5)) exposure on cardiovascular responses and remodeling and tested the effect of Rho kinase inhibition on these effects. C57BL/6 mice were exposed to concentrated ambient PM(2.5) or filtered air for 12 wk followed by a 14-day ANG II infusion in conjunction with fasudil, a Rho kinase antagonist, or placebo treatment. Blood pressure was monitored, followed by analysis of vascular function and ventricular remodeling indexes. PM(2.5) exposure potentiated ANG II-induced hypertension, and this effect was abolished by fasudil treatment. Cardiac and vascular RhoA activation was enhanced by PM(2.5) exposure along with increased expression of the guanine exchange factors (GEFs) PDZ-RhoGEF and p115 RhoGEF in PM(2.5)-exposed mice. Parallel with increased RhoA activation, PM(2.5) exposure increased ANG II induced cardiac hypertrophy and collagen deposition, with these increases being normalized by fasudil treatment. In conclusion, PM(2.5) potentiates cardiac remodeling in response to ANG II through RhoA/Rho kinase-dependent mechanisms. These findings have implications for the chronic cardiovascular health effects of air pollution. PMID- 19286944 TI - Increased O2 cost of basal metabolism and excitation-contraction coupling in hearts from type 2 diabetic mice. AB - We have reported previously that hearts from type 2 diabetic (db/db) mice show decreased cardiac efficiency due to increased work-independent myocardial O(2) consumption (unloaded MVo(2)), indicating higher O(2) use for nonmechanical processes such as basal metabolism (MVo(2)(BM)) and excitation-contraction coupling (MVo(2)(ECC)). Although alterations in cardiac metabolism and/or Ca(2+) handling may contribute to increased energy expenditure in diabetic hearts, direct measurements of the O(2) cost for these individual processes have not been determined. In this study, we 1) validate a procedure for measuring unloaded MVo(2) directly (MVo(2)(unloaded)) and for determining MVo(2)(BM) and MVo(2)(ECC) separately in isolated perfused mouse hearts and 2) determine O(2) cost for these processes in hearts from db/db mice. Unloaded MVo(2), extrapolated from the relationship between cardiac work (measured as pressure-volume area, PVA) and MVo(2), was found to correspond with MVo(2) measured directly in unloaded retrograde perfused hearts (MVo(2)(unloaded)). MVo(2) in K(+)-arrested hearts was defined as MVo(2)(BM); the difference between MVo(2)(unloaded) and MVo(2)(BM) represented MVo(2)(ECC). This procedure was validated by demonstrating that elevations in perfusate fatty acid (FA) and/or Ca(2+) concentrations resulted in changes in either MVo(2)(BM) and/or MVo(2)(ECC). The higher MVo(2)(unloaded) in db/db mice was due to both a higher MVo(2)(BM) and MVo(2)(ECC). Elevation of glucose and insulin decreased FA oxidation and reduced both MVo(2)(unloaded) and MVo(2)(BM). In conclusion, this study provides direct evidence that MVo(2)(BM) and MVo(2)(ECC) are elevated in diabetes and that acute metabolic interventions can have a therapeutic benefit in diabetic hearts due to a MVo(2)-lowering effect. PMID- 19286945 TI - Transfer function characteristics of the neural and peripheral arterial baroreflex arcs at rest and during postexercise muscle ischemia in humans. AB - Previous studies have demonstrated an increase in the arterial baroreflex (ABR) control of muscle sympathetic nerve activity (MSNA) during isolated activation of the muscle metaboreflex with postexercise muscle ischemia (PEMI). However, the increased ABR-MSNA control does not appear to manifest in an enhancement in the ABR control of arterial blood pressure (BP), suggesting alterations in the transduction of MSNA into a peripheral vascular response and a subsequent ABR mediated change in BP. Thus we examined the operating gains of the neural and peripheral arcs of the ABR and their interactive relationship at rest and during muscle metaboreflex activation. In nine healthy subjects, graded isolation of the muscle metaboreflex was achieved by PEMI following isometric handgrip performed at 15% and 30% maximal voluntary contraction (MVC). To obtain the sensitivities of the ABR neural and peripheral arcs, the transfer function gain from BP to MSNA and MSNA to femoral vascular conductance, respectively, was analyzed. No changes from rest were observed in the ABR neural or peripheral arcs during PEMI after 15% MVC handgrip. However, PEMI following 30% MVC handgrip increased the low frequency (LF) transfer function gain between BP and MSNA (ABR neural arc; +58 +/ 28%, P = 0.036), whereas the LF gain between MSNA and femoral vascular conductance (ABR peripheral arc) was decreased from rest (-36 +/- 8%, P = 0.017). These findings suggest that during high-intensity muscle metaboreflex activation an increased ABR gain of the neural arc appears to offset an attenuation of the peripheral arc gain to help maintain the overall ABR control of systemic BP. PMID- 19286946 TI - Is mean blood saturation a useful marker of tissue oxygenation? AB - Increasingly we are monitoring the distribution of oxygen through the microcirculation using optical techniques such as optical reflectance spectroscopy (ORS) and near-infrared spectroscopy. Mean blood oxygen saturation (S(mb)O(2)) and tissue oxygenation index measured by these two techniques, respectively, evoke a concept of the measurement of oxygen delivery to tissue. This study aims to establish whether S(mb)O(2) is an appropriate indicator of tissue oxygenation. Spontaneous fluctuations in S(mb)O(2) observed as changes in concentration of oxyhemoglobin ([HbO(2)]) and deoxyhemoglobin ([Hb]) were measured by ORS in the skin microcirculation of 30 healthy subjects (15 men, age 21-42 yr). Fourier analysis identified two distinctly different spontaneous falls in S(mb)O(2). The first type of swing, thought to be induced by fluctuations in arterial blood volume, resulted from the effects of respiration, endothelial, sympathetic, and myogenic activity. There was no apparent change in [Hb]. In contrast, a second type of swing resulted from a fall in [HbO(2)] accompanied by a rise in [Hb] and was only induced by endothelial and sympathetic activity. Thus the same fall in S(mb)O(2) can be induced by two distinct responses. A "type I" swing does not suggest an inadequacy in oxygen delivery whereas a "type II" swing may indicate a change in oxygen delivery from blood to tissue. S(mb)O(2) alone cannot therefore be accepted as a definitive marker of tissue oxygenation. PMID- 19286947 TI - Sex-dependent impairment of cardiac action potential conduction in type 1 diabetic rats. AB - The incidence of diabetes mellitus is increasing. Cardiac dysfunction often develops, resulting in diverse arrhythmias. These arise from ion channel remodeling or from altered speed and pattern of impulse propagation. Few studies have investigated impulse propagation in the diabetic heart. We previously showed a reduced conduction reserve in the diabetic heart, with associated changes in intercellular gap junctions. The present study investigated whether these effects are sex specific. Hearts from control and streptozotocin-diabetic male and female rats were used. Optical mapping was performed with the voltage-sensitive dye di-4 ANEPPS, using Langendorff-perfused hearts. Isolated ventricular cells and tissue sections were used for immunofluorescent labeling of the gap junction protein connexin43 (Cx43). The gap junction uncoupler heptanol (0.75 mM) or elevated K(+) (9 mM, to reduce cell excitability) produced significantly greater slowing of propagation in diabetic males than females. In ovariectomized diabetic females, 9 mM K(+) slowed conduction significantly more than in nonovariectomized females. The subcellular redistribution (lateralization) of the gap junction protein Cx43 was smaller in diabetic females. Pretreatment of diabetic males with the angiotensin-converting enzyme inhibitor quinapril reduced Cx43 lateralization and the effects of 9 mM K(+) on propagation. In conclusion, the slowing of cardiac impulse propagation in type 1 diabetes is smaller in female rats, partly due to the presence of female sex hormones. This difference is (partly) mediated by sex differences in activation of the cardiac renin-angiotensin system. PMID- 19286948 TI - Systemic injection of planktonic forms of mammalian-derived nanoparticles alters arterial response to injury in rabbits. AB - Experiments were designed to test the hypothesis that the systemic delivery of planktonic forms of nanoparticles (NPs) derived from calcified, diseased human tissue or bovine blood are transmissible particles that exacerbate arterial response to injury. New Zealand White rabbits in which the endothelium was mechanically removed from one carotid artery were injected intravenously with either saline (control), lipopolysaccharide (LPS; surrogate for subclinical infection), hydroxyapatite crystals (HA; surrogate for NP shell), HA crystals exposed to culture media, or planktonic forms of bovine- or human-derived NPs. Carotid arteries were monitored by ultrasonography for 5 wk and then removed for histological examination. Uninjured arteries from all animals in each group remained patent with a normal anatomy. Injured arteries from 6 of 11 animals injected with human-derived NPs occluded and/or calcified; none of the injured arteries from animals in the other groups occluded (n = 28; P < or = 0.05). Injured arteries of rabbits injected with LPS or HA crystals developed eccentric hyperplasia. Discontinuous internal elastic laminae and thinning media characterized arteries from animals injected with bovine-derived NPs or cultured HA crystals. In conclusion, the systemic administration of planktonic forms of human-derived NPs exacerbated arterial response to injury distinct from that of bovine-derived NPs and other inflammatory agents. PMID- 19286949 TI - Angiotensin II upregulates hypothalamic AT1 receptor expression in rats via the mitogen-activated protein kinase pathway. AB - ANG II type 1 receptors (AT(1)R) mediate most of the central effects of ANG II on cardiovascular function, fluid homeostasis, and sympathetic drive. The mechanisms regulating AT(1)R expression in the brain are unknown. In some tissues, the AT(1)R can be upregulated by prolonged exposure to ANG II. We examined the hypothesis that ANG II upregulates the AT(1)R in the brain by stimulating the intracellular mitogen-activated protein kinase (MAPK) signaling pathway. Using molecular and immunochemical approaches, we examined expression of the AT(1)R and phosphorylated MAPK in the paraventricular nucleus of the hypothalamus (PVN) and the subfornical organ (SFO) of rats receiving a chronic (4-wk) subcutaneous infusion of ANG II (0.6 microg/h) or saline (vehicle control), with or without concomitant (4-wk) intracerebroventricular (ICV) infusions of MAPK inhibitors or the AT(1)R blocker losartan. Subcutaneous infusion of ANG II markedly increased phosphorylation of MAPK and expression of AT(1)R mRNA and protein and AT(1)R-like immunoreactivity in the PVN and SFO. ANG II-induced AT(1)R expression was blocked by ICV infusion of the p44/42 MAPK inhibitor PD-98059 (0.025 microg/h) and the JNK inhibitor SP-600125 (0.125 microg/h), but not by the p38 MAPK inhibitor SB 203580 (0.125 microg/h). Upregulation of the AT(1)R in the PVN and SFO by peripheral ANG II was abolished by ICV losartan (10 microg/h). The data indicate that blood-borne ANG II upregulates brain AT(1)R by activating intracellular p44/42 MAPK and JNK signaling pathways. PMID- 19286950 TI - Prenatal cocaine exposure abolished ischemic preconditioning-induced protection in adult male rat hearts: role of PKCepsilon. AB - Prenatal cocaine exposure in rats resulted in decreased PKCepsilon protein expression in the heart of adult male but not female offspring. The present study determined its functional consequence of inhibiting cardioprotection mediated by ischemic preconditioning. Pregnant Sprague-Dawley rats were administered intraperitoneally saline or cocaine (30 mg.kg(-1).day(-1)) from day 15 to day 21 of gestational age. Hearts were isolated from 3-mo-old offspring and were subjected to ischemia and reperfusion injury in a Langendorff preparation, with or without prior ischemic preconditioning. Preischemic values of left ventricular function were the same between the saline control and cocaine-treated animals. Ischemic preconditioning of two episodes of 5-min ischemia significantly decreased infarct size and enhanced postischemic functional recovery of the left ventricle in the saline control animals. This ischemic preconditioning was associated with increased phospho-PKCepsilon, but not phospho-PKCdelta, levels and was blocked by a PKCepsilon translocation inhibitor peptide. Prenatal cocaine treatment abolished the ischemic preconditioning-mediated increase in phospho PKCepsilon and cardioprotection in the heart of male offspring. In contrast, the cardioprotective effect was fully maintained in female offspring that were exposed to cocaine before birth. The results suggest that prenatal cocaine exposure causes a sex-specific loss of cardioprotection by ischemic preconditioning in adult offspring, which is most likely due to fetal programming of PKCepsilon gene repression, resulting in a downregulation of PKCepsilon function in the heart of adult male offspring. PMID- 19286951 TI - The endothelial glycocalyx mediates shear-induced changes in hydraulic conductivity. AB - Recent in vitro and in vivo studies have reported fluid shear stress-induced increases in endothelial layer hydraulic conductivity (L(p)) that are mediated by an increased production of nitric oxide (NO). Other recent studies have shown that NO induction by shear stress is mediated by the glycocalyx that decorates the surface of endothelial cells. Here we find that a selective depletion of the major components of the glycocalyx with enzymes can block the shear stress induced response of L(p). Heparinase and hyaluronidase block shear-induced increases in L(p), which is consistent with their effects on NO production. But chondroitinase, which does not suppress shear-induced NO production, also inhibits shear-induced L(p). A further surprise is that treatment with the general proteolytic enzyme pronase does not suppress the shear L(p) response. We also find that heparinase does not alter baseline L(p) significantly, whereas chondroitinase, hyaluronidase, and pronase increase it significantly. PMID- 19286952 TI - Stretch-activated channel activation promotes early afterdepolarizations in rat ventricular myocytes under oxidative stress. AB - Mechanical stretch and oxidative stress have been shown to prolong action potential duration (APD) and produce early afterdepolarizations (EADs). Here, we developed a simulation model to study the role of stretch-activated channel (SAC) currents in triggering EADs in ventricular myocytes under oxidative stress. We adapted our coupling clamp circuit so that a model ionic current representing the actual SAC current was injected into ventricular myocytes and added as a real time current. This current was calculated as I(SAC) = G(SAC) * (V(m) - E(SAC)), where G(SAC) is the stretch-activated conductance, V(m) is the membrane potential, and E(SAC) is the reversal potential. In rat ventricular myocytes, application of G(SAC) did not produce sustained automaticity or EADs, although turn-on of G(SAC) did produce some transient automaticity at high levels of G(SAC). Exposure of myocytes to 100 microM H(2)O(2) induced significant APD prolongation and increase in intracellular Ca(2+) load and transient, but no EAD or sustained automaticity was generated in the absence of G(SAC). However, the combination of G(SAC) and H(2)O(2) consistently produced EADs at lower levels of G(SAC) (2.6 +/- 0.4 nS, n = 14, P < 0.05). Pacing myocytes at a faster rate further prolonged APD and promoted the development of EADs. SAC activation plays an important role in facilitating the development of EADs in ventricular myocytes under acute oxidative stress. This mechanism may contribute to the increased propensity to lethal ventricular arrhythmias seen in cardiomyopathies, where the myocardium stretch and oxidative stress generally coexist. PMID- 19286953 TI - Role of superoxide, nitric oxide, and peroxynitrite in doxorubicin-induced cell death in vivo and in vitro. AB - Doxorubicin (DOX) is a potent available antitumor agent; however, its clinical use is limited because of its cardiotoxicity. Cell death is a key component in DOX-induced cardiotoxicity, but its mechanisms are elusive. Here, we explore the role of superoxide, nitric oxide (NO), and peroxynitrite in DOX-induced cell death using both in vivo and in vitro models of cardiotoxicity. Western blot analysis, real-time PCR, immunohistochemistry, flow cytometry, fluorescent microscopy, and biochemical assays were used to determine the markers of apoptosis/necrosis and sources of NO and superoxide and their production. Left ventricular function was measured by a pressure-volume system. We demonstrated increases in myocardial apoptosis (caspase-3 cleavage/activity, cytochrome c release, and TUNEL), inducible NO synthase (iNOS) expression, mitochondrial superoxide generation, 3-nitrotyrosine (NT) formation, matrix metalloproteinase (MMP)-2/MMP-9 gene expression, poly(ADP-ribose) polymerase activation [without major changes in NAD(P)H oxidase isoform 1, NAD(P)H oxidase isoform 2, p22(phox), p40(phox), p47(phox), p67(phox), xanthine oxidase, endothelial NOS, and neuronal NOS expression] and decreases in myocardial contractility, catalase, and glutathione peroxidase activities 5 days after DOX treatment to mice. All these effects of DOX were markedly attenuated by peroxynitrite scavengers. Doxorubicin dose dependently increased mitochondrial superoxide and NT generation and apoptosis/necrosis in cardiac-derived H9c2 cells. DOX- or peroxynitrite-induced apoptosis/necrosis positively correlated with intracellular NT formation and could be abolished by peroxynitrite scavengers. DOX-induced cell death and NT formation were also attenuated by selective iNOS inhibitors or in iNOS knockout mice. Various NO donors when coadministered with DOX but not alone dramatically enhanced DOX-induced cell death with concomitant increased NT formation. DOX induced cell death was also attenuated by cell-permeable SOD but not by cell permeable catalase, the xanthine oxidase inhibitor allopurinol, or the NADPH oxidase inhibitors apocynine or diphenylene iodonium. Thus, peroxynitrite is a major trigger of DOX-induced cell death both in vivo and in vivo, and the modulation of the pathways leading to its generation or its effective neutralization can be of significant therapeutic benefit. PMID- 19286954 TI - Antidiabetic drug pioglitazone protects the heart via activation of PPAR-gamma receptors, PI3-kinase, Akt, and eNOS pathway in a rabbit model of myocardial infarction. AB - The insulin-sensitizing drug pioglitazone has been reported to be protective against myocardial infarction. However, its precise mechanism is unclear. Rabbits underwent 30 min of coronary occlusion followed by 48 h of reperfusion. Rabbits were assigned randomly to nine groups (n = 10 in each): the control group (fed a normal diet), pioglitazone group (fed diets containing 1 mg.kg(-1).day(-1) pioglitazone), pioglitazone + 5-hydroxydecanoic acid (HD) group [fed the pioglitazone diet + 5 mg/kg iv 5-HD, a mitochondrial ATP-sensitive K(+) (K(ATP)) channel blocker], pioglitazone + GW9662 group [fed the pioglitazone diet + 2 mg/kg iv GW9662, a peroxisome proliferator activated receptor (PPAR)-gamma antagonist], GW9662 group (fed a normal diet + iv GW9662), pioglitazone + wortmannin group [fed the pioglitazone diet + 0.6 mg/kg iv wortmannin, a phosphatidylinositol (PI)3-kinase inhibitor], wortmannin group (fed a normal diet + iv wortmannin), pioglitazone + nitro-l-arginine methyl ester (l-NAME) group [fed the pioglitazone diet + 10 mg/kg iv l-NAME, a nitric oxide synthase (NOS) inhibitor], and l-NAME group (fed a normal diet + iv l-NAME). All groups were fed the diets for 7 days. The risk area and nonrisk area of the left ventricle (LV) were separated by Evans blue dye, and the infarct area was determined by triphenyltetrazolium chloride staining. The infarct size was calculated as a percentage of the LV risk area. Western blotting was performed to assess levels of Akt and phospho-Akt and phospho-endothelial NOS (eNOS) in the myocardium following reperfusion. The infarct size was significantly smaller in the pioglitazone group (21 +/- 2%) than in the control group (43 +/- 3%). This effect was abolished by GW9662 (42 +/- 3%), wortmannin (40 +/- 3%), or l-NAME (42 +/- 7%) but not by 5-HD (24 +/- 5%). Western blotting showed higher levels of phospho Akt and phospho-eNOS in the pioglitazone group. Pioglitazone reduces the myocardial infarct size via activation of PPAR-gamma, PI3-kinase, Akt, and eNOS pathways, but not via opening the mitochondrial K(ATP) channel. Pioglitazone may be a novel strategy for the treatment of diabetes mellitus with coronary artery disease. PMID- 19286955 TI - Acidosis and ischemia increase cellular Ca2+ transient alternans and repolarization alternans susceptibility in the intact rat heart. AB - Cardiac cellular Ca(2+) transient (CaT) alternans and electrocardiographic T-wave alternans (TWA) often develop in myocardial ischemia, but the mechanisms for this relationship have not been elucidated. Acidosis is a major component of ischemia, but there is no direct evidence linking acidosis-induced cellular CaT alternans to ischemia-induced CaT alternans and TWA in whole heart. We used laser-scanning confocal microscopy to measure intracellular Ca(2+) (Ca(i)(2+)) cycling in individual myocytes of fluo-4 AM-loaded rat hearts and simultaneously recorded pseudo-ECGs to investigate changes in CaTs and late-phase repolarization, respectively, during baseline and rapid pacing under control and either globally acidic or globally ischemic conditions. Acidosis (hypercapnia; pH 6.6) increased diastolic Ca(i)(2+) levels, prolonged CaT duration, and shifted to slower heart rates both the development of pacing-induced acidosis-induced CaT alternans (both concordant and discordant) and of repolarization alternans (RPA, a measure of TWA in rat ECGs). The magnitudes of these shifts were equivalent for both CaT alternans and RPA, suggesting a close association between them. Nearly identical results were found in low-flow global ischemia. Additionally, ischemic preconditioning reduced the increased propensity for CaT alternans and RPA development and was mimicked by preconditioning by acidosis alone. Our results demonstrate that global acidosis or ischemia modifies Ca(i)(2+) cycling in myocytes such that the diastolic Ca(i)(2+) rises and the cellular CaT duration is prolonged, causing spatially concordant as well as spatially discordant cellular CaT alternans to develop at slower heart rates than in controls. Since RPA also developed at slower heart rates, our results suggest that acidosis is a major contributor to CaT alternans, which underlies the proarrhythmic state induced by myocardial ischemia and therefore may play a role in its modulation and prevention. PMID- 19286956 TI - Detrimental effect of combined exercise training and eNOS overexpression on cardiac function after myocardial infarction. AB - It has been reported that exercise after myocardial infarction (MI) attenuates left ventricular (LV) pump dysfunction by normalization of myofilament function. This benefit could be due to an exercise-induced upregulation of endothelial nitric oxide synthase (eNOS) expression and activity. Consequently, we first tested the hypothesis that the effects of exercise after MI can be mimicked by elevated eNOS expression using transgenic mice with overexpression of human eNOS (eNOSTg). Both exercise and eNOSTg attenuated LV remodeling and dysfunction after MI in mice and improved cardiomyocyte maximal force development (F(max)). However, only exercise training restored myofilament Ca(2+)-sensitivity and sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA)2a protein levels and improved the first derivative of LV pressure at 30 mmHg. Conversely, only eNOSTg improved survival. In view of these partly complementary actions, we subsequently tested the hypothesis that combining exercise and eNOSTg would provide additional protection against LV remodeling and dysfunction after MI. Unexpectedly, the combination of exercise and eNOSTg abolished the beneficial effects on LV remodeling and dysfunction of either treatment alone. The latter was likely due to perturbations in Ca(2+) homeostasis, as myofilament F(max) actually increased despite marked reductions in the phosphorylation status of several myofilament proteins, whereas the exercise-induced increases in SERCA2a protein levels were lost in eNOSTg mice. Antioxidant treatment with N-acetylcysteine or supplementation of tetrahydrobiopterin and l-arginine prevented these detrimental effects on LV function while partly restoring the phosphorylation status of myofilament proteins and further enhancing myofilament F(max). In conclusion, the combination of exercise and elevated eNOS expression abolished the cardioprotective effects of either treatment alone after MI, which appeared to be, at least in part, the result of increased oxidative stress secondary to eNOS "uncoupling." PMID- 19286957 TI - CBP and p300 in renin homeostasis: can they drive the fate? PMID- 19286958 TI - The shy Angeli and his elusive creature: the HNO route to vasodilation. PMID- 19286959 TI - Partial downregulation of junctin enhances cardiac calcium cycling without eliciting ventricular arrhythmias in mice. AB - Human failing hearts exhibit significant decreases in junctin expression levels with almost nondetectable levels, which may be associated with premature death, induced by lethal cardiac arrhythmias, based on mouse models. However, the specific contribution of junctin to the delayed afterdepolarizations has been difficult to delineate in the phase of increased Na(+)-Ca(2+) exchanger activity accompanying junctin ablation. Thus we characterized the heterozygous junctin deficient hearts, which expressed 54% of junctin levels and similar increases in Na(+)-Ca(2+) exchanger activity, as the null model. Cardiac contractile parameters, Ca(2+) transients, and sarcoplasmic reticulum Ca(2+) content were significantly increased in junctin heterozygous hearts, although they did not reach the levels of null hearts. However, Ca(2+) spark properties were not altered in heterozygous cardiomyocytes, compared with wild-types, and there were no aftercontractions elicited by the increased frequency of stimulation in the presence of isoproterenol, unlike the junctin-deficient cells. Furthermore, heterozygous mice did not exhibit an increased susceptibility to arrhythmia upon catecholamine challenge in vivo, and there were no premature deaths up to 1 yr of age. These findings suggest that a partial downregulation of junctin enhances sarcoplasmic reticulum Ca(2+) cycling but does not elicit cardiac arrhythmias even in the context of increased Na(+)-Ca(2+) exchanger activity. PMID- 19286960 TI - HERG biosynthesis: the positive influence of negative charge. PMID- 19286961 TI - ERK phosphorylation mediates sildenafil-induced myocardial protection against ischemia-reperfusion injury in mice. AB - Sildenafil, a selective inhibitor of phosphodiesterase type 5, induces powerful protection against myocardial ischemia-reperfusion injury through activation of cGMP-dependent protein kinase (PKG). We further hypothesized that PKG-dependent activation of survival kinase ERK may play a causative role in sildenafil-induced cardioprotection via induction of endothelial nitric oxide synthase (eNOS)/inducible nitric oxide synthase (iNOS) and Bcl-2. Our results show that acute intracoronary infusion of sildenafil in Langendorff isolated mouse hearts before global ischemia-reperfusion significantly reduced myocardial infarct size (from 29.4 +/- 2.4% to 15.9 +/- 3.0%; P < 0.05). Cotreatment with ERK inhibitor PD98059 abrogated sildenafil-induced protection (31.8 +/- 4.4%). To further evaluate the role of ERK in delayed cardioprotection, mice were treated with sildenafil (ip) 24 h before global ischemia-reperfusion. PD98059 was administered (ip) 30 min before sildenafil treatment. Infarct size was reduced from 27.6 +/- 3.3% in controls to 7.1 +/- 1.5% in sildenafil-treated mice (P < 0.05). The delayed protective effect of sildenafil was also abolished by PD98059 (22.5 +/- 2.3%). Western blots revealed that sildenafil significantly increased phosphorylation of ERK1/2 and GSK-3beta and induced iNOS, eNOS, Bcl-2, and PKG activity in the heart 24 h after treatment. PD98059 inhibited the enhanced expression of iNOS, eNOS, and Bcl-2 and the phosphorylation of GSK-3beta. PD98059 had no effect on the sildenafil-induced activation of PKG. We conclude that these studies provide first direct evidence that PKG-dependent ERK phosphorylation is indispensable for the induction of eNOS/iNOS and Bcl-2 and the resulting cardioprotection by sildenafil. PMID- 19286962 TI - Lack of S100A1 in mice confers a gender-dependent hypertensive phenotype and increased mortality after myocardial infarction. AB - S100A1 is a small Ca(2+)-binding protein expressed in the myocardium and blood vessels that is downregulated in the diseased heart and plays a role in the regulation of cardiac muscle Ca(2+) homeostasis and contractility. To understand its physiological role under basal conditions and after myocardial infarction (MI), we used a mouse strain with targeted deletion of the S100A1 gene [S100A1 knockout (KO) mice]. We compared 49 wild-type (WT) and 56 S100A1 KO mice (6-8 wk old) over 28 days after MI with sham-operated controls. We also examined the effect of S100A1 deficiency on vascular function of isolated blood vessels. S100A1 KO mice demonstrated worse survival compared with WT mice (21% vs. 69%, respectively, P < 0.001). Hemodynamic evaluation revealed a higher mean arterial pressure (MAP) in sham-operated KO animals compared with WT animals (99 +/- 4 vs. 77 +/- 3 mmHg, respectively, P < 0.001) that persisted in both groups after MI (86 +/- 2 vs. 66 +/- 4 mmHg, respectively, P < 0.001). Sham-operated male S100A1 KO mice had higher MAP than female KO mice (122 +/- 5 vs. 93 +/- 3 mmHg, respectively P < 0.05) and reduced survival after MI (4% vs. 27%, respectively, P < 0.05). In isolated aortas and mesenteric arteries, ACh-evoked vasodilatation in KO mice was significantly reduced compared with WT mice (P < 0.05). Nitric oxide production was reduced in endothelial cells isolated from KO mice. Thus, absence of S100A1 results in acute functional impairment and high mortality after MI associated with a gender-specific hypertensive phenotype. S100A1 appears to play a role in the endothelium-dependent regulation of blood pressure. PMID- 19286963 TI - Contribution of nerve growth factor to augmented TRPV1 responses of muscle sensory neurons by femoral artery occlusion. AB - In rats, hindlimb muscle ischemia induced by femoral artery occlusion augments the sympathetic nervous response to stimulation of transient receptor potential vanilloid type 1 (TRPV1) by injection of capsaicin into the arterial blood supply of the hindlimb muscles. The enhanced sympathetic response is due to alterations in TRPV1 receptor expression and its responsiveness in sensory neurons. The underlying mechanism by which TRPV1 receptor responses are increased after muscle vascular insufficiency/ischemia is unclear. In this report we tested the hypothesis that muscle ischemia elevates nerve growth factor (NGF) levels in primary afferent neurons, thereby increasing TRPV1 responsiveness. Muscle vascular insufficiency induced by the femoral artery ligation significantly increased NGF in the dorsal root ganglion (DRG) compared with sham controls. Furthermore, when NGF was infused in the hindlimb muscles of healthy rats (72 h using an osmotic minipump), the magnitude of the DRG neuron response to capsaicin was augmented (5.4 +/- 0.54 nA with NGF infusion vs. 3.0 +/- 0.17 nA in control; P < 0.05). With the addition of NGF in the culture dish containing the DRG neurons, the magnitude of the DRG neuron response to capsaicin was greater (6.4 +/- 0.27 nA; P < 0.05 vs. control) than that seen in control (2.9 +/- 0.16 nA). Note that this NGF effect was seen in isolectin B(4)-negative DRG neurons, a group of thin fiber nerves that contain neuropeptides and depend on NGF for survival. These data suggest that NGF affects a selective subpopulation of the afferent neurons in mediating augmented TRPV1 responses after femoral artery occlusion. PMID- 19286964 TI - Involvement of NO and MEK/ERK pathway in enhancement of endothelin-1-induced mesenteric artery contraction in later-stage type 2 diabetic Goto-Kakizaki rat. AB - Endothelin (ET)-1 is a likely candidate for a key role in diabetic vascular complications. However, no abnormalities in the vascular responsiveness to ET-1 have been identified in the chronic stage of type 2 diabetes. Our goal was to look for abnormalities in the roles played by ET receptors (ET(A) and ET(B)) in the mesenteric artery of the type 2 diabetic Goto-Kakizaki (GK) rat and to identify the molecular mechanisms involved. Using mesenteric arteries from later stage (32-38 wk old) individuals, we compared the ET-1-induced contraction and the relaxation induced by the selective ET(B) receptor agonist IRL1620 between GK rats and control Wistar rats. Mesenteric artery ERK activity and the protein expressions for ET receptors and MEK were also measured. In GK rats (vs. age matched Wistar rats), we found as follows. 1) The ET-1-induced contraction was greater and was attenuated by BQ-123 (ET(A) antagonist) but not by BQ-788 (ET(B) antagonist). In the controls, BQ-788 augmented this contraction. 2) Both the relaxation and nitric oxide (NO) production induced by IRL1620 were reduced. 3) ET-1-induced contraction was enhanced by N(G)-nitro-l-arginine (l-NNA; NO synthase inhibitor) but suppressed by sodium nitroprusside (NO donor). 4) The enhanced ET-1-induced contraction was reduced by MEK/ERK pathway inhibitors (PD 98059 or U0126). 5) ET-1-stimulated ERK activation was increased, as were the ET(A) and MEK1/2 protein expressions. 6) Mesenteric ET-1 content was increased. These results suggest that upregulation of ET(A), a defect in ET(B)-mediated NO signaling, and activation of the MEK/ERK pathway together represent a likely mechanism mediating the hyperreactivity to ET-1 examined in this study. PMID- 19286965 TI - Atherosclerotic plaque disruption induced by stress and lipopolysaccharide in apolipoprotein E knockout mice. AB - To establish an animal model with disruptions of atherosclerotic plaques, 96 male apolipoprotein E knockout (apoE(-/-)) mice were randomly divided into stress, lipopolysaccharide (LPS), stress+LPS, and control groups (n = 24 each). All mice were fed a high-fat diet throughout the experiment, and carotid atherosclerotic lesions were induced by placement of a constrictive perivascular collar. Four weeks after surgery, mice in the LPS and stress+LPS groups were intraperitoneally injected with LPS (1 mg/kg twice per week for 8 wk). Eight weeks after surgery, mice in the stress and stress+LPS groups were treated with intermittent physical stress (electric foot shock and noise stimulation) for 4 wk. Morphological analysis revealed a plaque disruption rate of 16.7% in control, 34.8% in LPS, 54.2% in stress, and 60.9% in stress+LPS groups. The disruption rates in stress and stress+LPS groups were both significantly higher than those of controls (P = 0.007 and P = 0.002, respectively). Luminal thrombosis secondary to plaque disruption was observed only in the stress+LPS group. Both stress and LPS stimulation significantly decreased fibrous cap thickness and increased macrophage and lipid contents in plaques. Moreover, the combination of stress and LPS stimulation further lowered cap thickness and enhanced accumulation of macrophages and expression of inflammatory cytokines and matrix metalloproteinases. Stress activated the sympathetic nervous system, as manifested by increased blood pressure and flow velocity. Plasma fibrinogen levels were remarkably elevated in the stress and stress+LPS groups. In conclusion, stress- and LPS-costimulated apoE(-/-) mice provide a useful model for studies of plaque vulnerability and interventions. PMID- 19286966 TI - Opposites attract: some phytochromes do not form homodimers. PMID- 19286967 TI - Obligate heterodimerization of Arabidopsis phytochromes C and E and interaction with the PIF3 basic helix-loop-helix transcription factor. AB - Phytochromes are dimeric chromoproteins that regulate plant responses to red (R) and far-red (FR) light. The Arabidopsis thaliana genome encodes five phytochrome apoproteins: type I phyA mediates responses to FR, and type II phyB-phyE mediate shade avoidance and classical R/FR-reversible responses. In this study, we describe the complete in vivo complement of homodimeric and heterodimeric type II phytochromes. Unexpectedly, phyC and phyE do not homodimerize and are present in seedlings only as heterodimers with phyB and phyD. Roles in light regulation of hypocotyl length, leaf area, and flowering time are demonstrated for heterodimeric phytochromes containing phyC or phyE. Heterodimers of phyC and chromophoreless phyB are inactive, indicating that phyC subunits require spectrally intact dimer partners to be active themselves. Consistent with the obligate heterodimerization of phyC and phyE, phyC is made unstable by removal of its phyB binding partner, and overexpression of phyE results in accumulation of phyE monomers. Following a pulse of red light, phyA, phyB, phyC, and phyD interact in vivo with the PHYTOCHROME INTERACTING FACTOR3 basic helix-loop-helix transcription factor, and this interaction is FR reversible. Therefore, most or all of the type I and type II phytochromes, including heterodimeric forms, appear to function through PIF-mediated pathways. These findings link an unanticipated diversity of plant R/FR photoreceptor structures to established phytochrome signaling mechanisms. PMID- 19286968 TI - A chloroplastic UDP-glucose pyrophosphorylase from Arabidopsis is the committed enzyme for the first step of sulfolipid biosynthesis. AB - Plants synthesize a sulfur-containing lipid, sulfoquinovosyldiacylglycerol, which is one of three nonphosphorus glycerolipids that provide the bulk of the structural lipids in photosynthetic membranes. Here, the identification of a novel gene, UDP-glucose pyrophosphorylase3 (UGP3), required for sulfolipid biosynthesis is described. Transcriptome coexpression analysis demonstrated highly correlated expression of UGP3 with known genes for sulfolipid biosynthesis in Arabidopsis thaliana. Liquid chromatography-mass spectrometry analysis of leaf lipids in two Arabidopsis ugp3 mutants revealed that no sulfolipid was accumulated in these mutants, indicating the participation of UGP3 in sulfolipid biosynthesis. From the deduced amino acid sequence, UGP3 was presumed to be a UDP glucose pyrophosphorylase (UGPase) involved in the generation of UDP-glucose, serving as the precursor of the polar head of sulfolipid. Recombinant UGP3 was able to catalyze the formation of UDP-glucose from glucose-1-phosphate and UTP. A transient assay using fluorescence fusion proteins and UGPase activity in isolated chloroplasts indicated chloroplastic localization of UGP3. The transcription level of UGP3 was increased by phosphate starvation. A comparative genomics study on UGP3 homologs across different plant species suggested the structural and functional conservation of the proteins and, thus, a committing role for UGP3 in sulfolipid synthesis. PMID- 19286969 TI - HISTONE MONOUBIQUITINATION1 interacts with a subunit of the mediator complex and regulates defense against necrotrophic fungal pathogens in Arabidopsis. AB - This work examines the role of the Arabidopsis thaliana RING E3 ligase, HISTONE MONOUBIQUITINATION1 (HUB1) in disease resistance. Loss-of-function alleles of HUB1 show increased susceptibility to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola, whereas HUB1 overexpression conferred resistance to B. cinerea. By contrast, responses to the bacterial pathogen Pseudomonas syringae are unaltered in hub1 plants. hub1 mutants have thinner cell walls but increased callose around an infection site. HUB1 acts independently of jasmonate, but ethylene (ET) responses and salicylate modulate the resistance of hub1 mutants to necrotrophic fungi. The ET response factor ETHYLENE INSENSITIVE2 is epistatic to HUB1 for A. brassicicola resistance but additive to HUB1 for B. cinerea resistance. HUB1 interacts with MED21, a subunit of the Arabidopsis Mediator, a conserved complex that regulates RNA polymerase II. RNA interference lines with reduced MED21 expression are highly susceptible to A. brassicicola and B. cinerea, whereas T-DNA insertion alleles are embryonic lethal, suggesting an essential role for MED21. However, HUB1-mediated histone H2B modification is independent of histone H3 and DNA methylation. In sum, histone H2B monoubiquitination is an important chromatin modification with regulatory roles in plant defense against necrotrophic fungi most likely through modulation of gene expression. PMID- 19286970 TI - Retroposon analysis and recent geological data suggest near-simultaneous divergence of the three superorders of mammals. AB - As a consequence of recent developments in molecular phylogenomics, all extant orders of placental mammals have been grouped into 3 lineages: Afrotheria, Xenarthra, and Boreotheria, which originated in Africa, South America, and Laurasia, respectively. Despite this advancement, the order of divergence of these 3 lineages remains unresolved. Here, we performed extensive retroposon analysis with mammalian genomic data. Surprisingly, we identified a similar number of informative retroposon loci that support each of 3 possible phylogenetic hypotheses: the basal position for Afrotheria (22 loci), Xenarthra (25 loci), and Boreotheria (21 loci). This result indicates that the divergence of the placental common ancestor into the 3 lineages occurred nearly simultaneously. Thus, we examined whether these molecular data could be integrated into the geological context by incorporating recent geological data. We obtained firm evidence that complete separation of Gondwana into Africa and South America occurred 120 +/- 10 Ma. Accordingly, the previous reported time frame (division of Pangea into Gondwana and Laurasia at 148-138 Ma and division of Gondwana at 105 Ma) cannot be used to validate mammalian divergence order. Instead, we use our retroposon results and the recent geological data to propose that near-simultaneous divisions of continents leading to isolated Africa, South America, and Laurasia caused nearly concomitant divergence of the ancient placental ancestor into 3 lineages, Afrotheria, Xenarthra, and Boreotheria, approximately 120 Ma. PMID- 19286971 TI - Sympathetic nervous system control of anti-influenza CD8+ T cell responses. AB - Despite the longstanding appreciation of communication between the nervous and the immune systems, the nature and significance of these interactions to immunity remain enigmatic. Here, we show that 6-hydroxydopamine-mediated ablation of the mouse peripheral sympathetic nervous system increases primary CD8(+) T cell responses to viral and cellular antigens presented by direct priming or cross priming. The sympathetic nervous system also suppresses antiviral CD4(+) T cell responses, but this is not required for suppressing CD8(+) T cell responses. Adoptive transfer experiments indicate that enhanced CD8(+) responses do not result from permanent alterations in CD8(+) T cell function in sympathectomized mice. Rather, additional findings suggest that the sympathetic nervous system tempers the capacity of antigen-presenting cells to activate naive CD8(+) T cells. We also show that antiviral CD8(+) T cell responses are enhanced by administration of a beta(2) (but not beta(1) or alpha) adrenergic antagonist. These findings demonstrate a critical role for the sympathetic nervous system in limiting CD8(+) T cell responses and indicate that CD8(+) T cell responses may be altered in patients using beta-blockers, one of the most widely prescribed classes of drugs. PMID- 19286972 TI - Bilirubin and glutathione have complementary antioxidant and cytoprotective roles. AB - Glutathione (GSH) and bilirubin are prominent endogenous antioxidant cytoprotectants. Despite tissue levels that are thousands of times lower than GSH, bilirubin is effective because of the biosynthetic cycle wherein it is generated from biliverdin by biliverdin reductase (BVR). When bilirubin acts as an antioxidant, it is oxidized to biliverdin, which is immediately reduced by BVR to bilirubin. Why does the body employ both of these 2 distinct antioxidant systems? We show that the water-soluble GSH primarily protects water soluble proteins, whereas the lipophilic bilirubin protects lipids from oxidation. Mice with deletion of heme oxygenase-2, which generates biliverdin, display greater lipid than protein oxidation, while the reverse holds for GSH depletion. RNA interference depletion of BVR increases oxidation of lipids more than protein. Depletion of BVR or GSH augments cell death in an oxidant-specific fashion. PMID- 19286974 TI - Retro-nasal aroma release depends on both subject and product differences: a link to food intake regulation? AB - It is hypothesized that differences in the extent of retro-nasal aroma release during consumption may be 1 of the reasons that people vary in their satiation characteristics. Using real-time atmospheric pressure chemical ionization mass spectrometry (APcI-MS), in vivo retro-nasal aroma release was determined for 30 subjects consuming 9 different food products, varying in physical structure (i.e., [semi]liquid and solid food products). Additionally, for a subset of the subjects ad libitum food intake was measured. Retro-nasal aroma release intensity and profile morphology appeared to be subject specific and relatively independent of the type of food product subjects consumed. A subject who was observed as having a relatively high retro-nasal aroma release intensity for a (semi)liquid food product also appeared to have a relatively high retro-nasal aroma release intensity for a solid food product. However, for all subjects, there were absolute differences between food products in the extent of retro-nasal aroma release comparing (semi)liquid and solid food products. This implies that the extent of retro-nasal aroma release is a valid physiological feature that characterizes any individual. Interestingly, a negative trend was observed between extent of retro-nasal aroma release and amount of ad libitum food intake (P = 0.07). This may have implications for the regulation of food intake. PMID- 19286975 TI - Alterations in oxygen consumption, respiratory quotient, and heat production in long-lived GHRKO and Ames dwarf mice, and short-lived bGH transgenic mice. AB - Growth hormone (GH) signaling influences longevity in mice, with decreased GH signaling associated with longer life span and increased GH signaling with shortened life span. A proposed mechanism through which GH signaling influences life span postulates that decreased GH signaling lowers metabolic rate, thus slowing aging by decreasing production of damaging free radicals. The influence of altered GH signaling on metabolism was tested by monitoring oxygen consumption (VO(2)), respiratory quotient (RQ), and heat production in long-lived GH receptor knockout (GHRKO) and Ames dwarf mice, and short-lived bovine GH-overexpressing transgenic (bGH TG) mice. Intriguingly, both GHRKO and Ames dwarf mice have increased VO(2) and heat per gram body weight, and decreased RQ, whereas bGH TG mice have decreased VO(2) and heat per gram body weight and increased RQ. In conclusion, decreased GH signaling associates with increased metabolism per body weight and may beneficially affect mitochondrial flexibility by increasing the capacity for fat oxidation; generally, GH excess produces opposite metabolic effects. PMID- 19286976 TI - Provisional clinical opinion. PMID- 19286977 TI - Electrophilic peroxisome proliferator-activated receptor-gamma ligands have potent antifibrotic effects in human lung fibroblasts. AB - Pulmonary fibrosis is a progressive scarring disease with no effective treatment. Transforming growth factor (TGF)-beta is up-regulated in fibrotic diseases, where it stimulates differentiation of fibroblasts to myofibroblasts and production of excess extracellular matrix. Peroxisome proliferator-activated receptor (PPAR) gamma is a transcription factor that regulates adipogenesis, insulin sensitization, and inflammation. We report here that a novel PPARgamma ligand, 2 cyano-3,12-dioxoolean-1,9-dien-28-oic acid (CDDO), is a potent inhibitor of TGF beta-stimulated differentiation of human lung fibroblasts to myofibroblasts, and suppresses up-regulation of alpha-smooth muscle actin, fibronectin, collagen, and the novel myofibroblast marker, calponin. The inhibitory concentration causing a 50% decrease in aSMA for CDDO was 20-fold lower than the endogenous PPARgamma ligand, 15-deoxy-Delta(12,14)-prostaglandin J(2) (15 d-PGJ(2)), and 400-fold lower than the synthetic ligand, rosiglitazone. Pharmacologic and genetic approaches were used to demonstrate that CDDO mediates its activity via a PPARgamma-independent pathway. CDDO and 15 d-PGJ(2) contain an alpha/beta unsaturated ketone, which acts as an electrophilic center that can form covalent bonds with cellular proteins. Prostaglandin A(1) and diphenyl diselenide, both strong electrophiles, also inhibit myofibroblast differentiation, but a structural analog of 15 d-PGJ(2) lacking the electrophilic center is much less potent. CDDO does not alter TGF-beta-induced Smad or AP-1 signaling, but does inhibit acetylation of CREB binding protein/p300, a critical coactivator in the transcriptional regulation of TGF-beta-responsive genes. Overall, these data indicate that certain PPARgamma ligands, and other small molecules with electrophilic centers, are potent inhibitors of critical TGF-beta-mediated profibrogenic activities through pathways independent of PPARgamma. As the inhibitory concentration causing a 50% decrease in aSMA for CDDO is 400-fold lower than that in rosiglitazone, the translational potential of CDDO for treatment of fibrotic diseases is high. PMID- 19286978 TI - Ubiquitination participates in the lysosomal degradation of Na,K-ATPase in steady state conditions. AB - The alveolar epithelial cell (AEC) Na,K-ATPase contributes to vectorial Na(+) transport and plays an important role in keeping the lungs free of edema. We determined, by cell surface labeling with biotin and immunofluorescence, that approximately 30% of total Na,K-ATPase is at the plasma membrane of AEC in steady state conditions. The half-life of the plasma membrane Na,K-ATPase was about 4 hours, and the incorporation of new Na,K-ATPase to the plasma membrane was Brefeldin A sensitive. Both protein kinase C (PKC) inhibition with bisindolylmaleimide (10 microM) and infection with an adenovirus expressing dominant-negative PKCzeta prevented Na,K-ATPase degradation. In cells expressing the Na,K-ATPase alpha1-subunit lacking the PKC phosphorylation sites, the plasma membrane Na,K-ATPase had a moderate increase in half-life. We also found that the Na,K-ATPase was ubiquitinated in steady-state conditions and that proteasomal inhibitors prevented its degradation. Interestingly, mutation of the four lysines described to be necessary for ubiquitination and endocytosis of the Na,K-ATPase in injurious conditions did not have an effect on its half-life in steady-state conditions. Lysosomal inhibitors prevented Na,K-ATPase degradation, and co localization of Na,K-ATPase and lysosomes was found after labeling and chasing the plasma membrane Na,K-ATPase for 4 hours. Accordingly, we provide evidence suggesting that phosphorylation and ubiquitination are necessary for the steady state degradation of the plasma membrane Na,K-ATPase in the lysosomes in alveolar epithelial cells. PMID- 19286979 TI - The Pneumocystis meiotic PCRan1p kinase exhibits unique temperature-regulated activity. AB - Pneumocystis organisms are opportunistic fungal pathogens that cause significant pneumonia in immune-compromised hosts. Recent evidence has suggested that Pneumocystis carinii exists as separate mating types, and expresses and regulates proteins that govern meiosis and progression of the life cycle. This study was undertaken to investigate the activity of three life cycle-regulatory proteins in Pneumocystis, including two proteins essential in mating signaling, and a putative meiotic regulator, to determine the conditions under which they are most active. This study used V5/HIS-tagged PCRan1p, PCSte20p, and PCCbk1, purified from Saccharomyces cerevisiae strain, INVSC, as well as an in vitro Escherichia coli protein expression system to determine the optimal expression conditions of each protein in the presence of varying pH, temperature, and metal ions. These studies demonstrate an atypical enzymatic activity in PCRan1p, whereby the kinase was most active in the environmental conditions between 10 and 25 degrees C, compared with a dramatic reduction in activity above 30 degrees C, temperatures typically found within mammalian hosts. Circular dichroism and fluorescence spectroscopy suggest that PCRan1p becomes partially unfolded at 25 degrees C, leading to its most active conformation, whereas continued unfolding as temperature increases results in strongly suppressed activity. These studies suggest that, in vivo, while under conditions within the mammalian lung (typically 37 degrees C), PCRan1p kinase activity is largely suppressed, allowing better conditions for the activation of meiosis, whereas in ex vivo environments, PCRan1p kinase activity increases to arrest progression of the life cycle until conditions become more favorable. PMID- 19286980 TI - Phosphatidylethanolamine is required for normal cell morphology and cytokinesis in the fission yeast Schizosaccharomyces pombe. AB - To investigate the contributions of phosphatidylethanolamine to the growth and morphogenesis of the fission yeast Schizosaccharomyces pombe, we have characterized three predicted genes in this organism, designated psd1, psd2, and psd3, encoding phosphatidylserine decarboxylases, which catalyze the conversion of phosphatidylserine to phosphatidylethanolamine in both eukaryotic and prokaryotic organisms. S. pombe mutants carrying deletions in any one or two psd genes are viable in complex rich medium and synthetic defined minimal medium. However, mutants carrying deletions in all three psd genes (psd1-3Delta mutants) grow slowly in rich medium and are inviable in minimal medium, indicating that the psd1 to psd3 gene products share overlapping essential cellular functions. Supplementation of growth media with ethanolamine, which can be converted to phosphatidylethanolamine by the Kennedy pathway, restores growth to psd1-3Delta cells in minimal medium, indicating that phosphatidylethanolamine is essential for S. pombe cell growth. psd1-3Delta cells produce lower levels of phosphatidylethanolamine than wild-type cells, even in medium supplemented with ethanolamine, indicating that the Kennedy pathway can only partially compensate for the loss of phosphatidylserine decarboxylase activity in S. pombe. psd1 3Delta cells appear morphologically indistinguishable from wild-type S. pombe cells in medium supplemented with ethanolamine, but when cultured in nonsupplemented medium, they produce high frequencies of abnormally shaped cells as well as cells exhibiting severe septation defects, including multiple, mispositioned, deformed, and misoriented septa. Our results demonstrate that phosphatidylethanolamine is essential for cell growth and for normal cytokinesis and cellular morphogenesis in S. pombe, and they illustrate the usefulness of this model eukaryote for investigating potentially conserved biological and molecular functions of phosphatidylethanolamine. PMID- 19286981 TI - Lessons from fungal F-box proteins. PMID- 19286982 TI - Altering sphingolipid metabolism in Saccharomyces cerevisiae cells lacking the amphiphysin ortholog Rvs161 reinitiates sugar transporter endocytosis. AB - Amphiphysins are proteins thought to be involved in synaptic vesicle endocytosis. Amphiphysins share a common BAR domain, which can sense and/or bend membranes, and this function is believed to be essential for endocytosis. Saccharomyces cerevisiae cells lacking the amphiphysin ortholog Rvs161 are inviable when starved for glucose. Altering sphingolipid levels in rvs161 cells remediates this defect, but how lipid changes suppress remains to be elucidated. Here, we show that the sugar starvation-induced death of rvs161 cells extends to other fermentable sugar carbon sources, and the loss of sphingolipid metabolism suppresses these defects. In all cases, rvs161 cells respond to the starvation signal, elicit the appropriate transcriptional response, and properly localize the requisite sugar transporter(s). However, Rvs161 is required for transporter endocytosis. rvs161 cells accumulate transporters at the plasma membrane under conditions normally resulting in their endocytosis and degradation. Transporter endocytosis requires the endocytosis (endo) domain of Rvs161. Altering sphingolipid metabolism by deleting the very-long-chain fatty acid elongase SUR4 reinitiates transporter endocytosis in rvs161 and rvs161 endo(-) cells. The sphingolipid-dependent reinitiation of endocytosis requires the ubiquitin regulating factors Doa1, Doa4, and Rsp5. In the case of Doa1, the phospholipase A(2) family ubiquitin binding motif is dispensable. Moreover, the conserved AAA ATPase Cdc48 and its accessory proteins Shp1 and Ufd1 are required. Finally, rvs161 cells accumulate monoubiquitin, and this defect is remediated by the loss of SUR4. These results show that defects in sphingolipid metabolism result in the reinitiation of ubiquitin-dependent sugar transporter endocytosis and suggest that this event is necessary for suppressing the nutrient starvation-induced death of rvs161 cells. PMID- 19286983 TI - Live-Cell imaging and measurement of intracellular pH in filamentous fungi using a genetically encoded ratiometric probe. AB - A novel, genetically encoded, ratiometric pH probe (RaVC) was constructed to image and measure intracellular pH in living hyphae of Aspergillus niger. RaVC is a chimeric protein based on the pH-sensitive probe pHluorin, which was partially codon optimized for expression in Aspergillus. Intracellular pH imaging and measurement was performed by simultaneous, dual-excitation confocal ratio imaging. The mean cytoplasmic pH measured was 7.4 to 7.7 based on calibrating RaVC in situ within nigericin-treated hyphae. Pronounced, longitudinal cytoplasmic pH gradients were not observed in the apical 20 microm of actively growing hyphae at the periphery of 18-h-old colonies. The cytoplasmic pH remained unchanged after prolonged growth in buffered medium with pH values between 2.5 or 9.5. Sudden changes in external pH significantly changed cytoplasmic pH by <1.3 pH units, but it returned to its original value within 20 min following treatment. The weak acid and antifungal food preservative sorbic acid caused prolonged, concentration-dependent intracellular acidification. The inhibition of ATPases with N-ethylmaleimide, dicychlohexylcarbodimide, or sodium azide caused the cytoplasmic pH to decrease by <1 pH unit. Treatment with the protonophore carbonyl cyanide m-chlorophenylhydrazone or cyanide p-(trifluoromethoxy) phenylhydrazone reduced the cytoplasmic pH by <1 pH unit. In older hyphae from 32 h-old cultures, RaVC became sequestered within large vacuoles, which were shown to have pH values between 6.2 and 6.5. Overall, our study demonstrates that RaVC is an excellent probe for visualizing and quantifying intracellular pH in living fungal hyphae. PMID- 19286984 TI - Microtubule-nucleus interactions in Dictyostelium discoideum mediated by central motor kinesins. AB - Kinesins are a diverse superfamily of motor proteins that drive organelles and other microtubule-based movements in eukaryotic cells. These motors play important roles in multiple events during both interphase and cell division. Dictyostelium discoideum contains 13 kinesin motors, 12 of which are grouped into nine families, plus one orphan. Functions for 11 of the 13 motors have been previously investigated; we address here the activities of the two remaining kinesins, both isoforms with central motor domains. Kif6 (of the kinesin-13 family) appears to be essential for cell viability. The partial knockdown of Kif6 with RNA interference generates mitotic defects (lagging chromosomes and aberrant spindle assemblies) that are consistent with kinesin-13 disruptions in other organisms. However, the orphan motor Kif9 participates in a completely novel kinesin activity, one that maintains a connection between the microtubule organizing center (MTOC) and nucleus during interphase. kif9 null cell growth is impaired, and the MTOC appears to disconnect from its normally tight nuclear linkage. Mitotic spindles elongate in a normal fashion in kif9(-) cells, but we hypothesize that this kinesin is important for positioning the MTOC into the nuclear envelope during prophase. This function would be significant for the early steps of cell division and also may play a role in regulating centrosome replication. PMID- 19286985 TI - Fungal functional genomics: tunable knockout-knock-in expression and tagging strategies. AB - Strategies for promoting high-efficiency homologous gene replacement have been developed and adopted for many filamentous fungal species. The next generation of analysis requires the ability to manipulate gene expression and to tag genes expressed from their endogenous loci. Here we present a suite of molecular tools that provide versatile solutions for fungal high-throughput functional genomics studies based on locus-specific modification of any target gene. Additionally, case studies illustrate caveats to presumed overexpression constructs. A tunable expression system and different tagging strategies can provide valuable phenotypic information for uncharacterized genes and facilitate the analysis of essential loci, an emerging problem in systematic deletion studies of haploid organisms. PMID- 19286986 TI - Type II NADH dehydrogenase inhibitor 1-hydroxy-2-dodecyl-4(1H)quinolone leads to collapse of mitochondrial inner-membrane potential and ATP depletion in Toxoplasma gondii. AB - The apicomplexan parasite Toxoplasma gondii expresses type II NADH dehydrogenases (NDH2s) instead of canonical complex I at the inner mitochondrial membrane. These non-proton-pumping enzymes are considered to be promising drug targets due to their absence in mammalian cells. We recently showed by inhibition kinetics that T. gondii NDH2-I is a target of the quinolone-like compound 1-hydroxy-2-dodecyl 4(1H)quinolone (HDQ), which inhibits T. gondii replication in the nanomolar range. In this study, the cationic fluorescent probes Mitotracker and DiOC(6)(3) (3,3'-dihexyloxacarbocyanine iodine) were used to monitor the influence of HDQ on the mitochondrial inner membrane potential (Delta Psi m) in T. gondii. Real-time imaging revealed that nanomolar HDQ concentrations led to a Delta Psi m collapse within minutes, which is followed by severe ATP depletions of 30% after 1 h and 70% after 24 h. Delta Psi m depolarization was attenuated when substrates for other dehydrogenases that can donate electrons to ubiquinone were added to digitonin-permeabilized cells or when infected cultures were treated with the F(o)-ATPase inhibitor oligomycin. A prolonged treatment with sublethal concentrations of HDQ induced differentiation into bradyzoites. This dormant stage is likely to be less dependent on the Delta Psi m, since Delta Psi m positive parasites were found at a significantly lower frequency in alkaline-pH induced bradyzoites than in tachyzoites. Together, our studies reveal that oxidative phosphorylation is essential for maintaining the ATP level in the fast growing tachyzoite stage and that HDQ interferes with this pathway by inhibiting the electron transport chain at the level of ubiquinone reduction. PMID- 19286987 TI - Differential roles of the ChiB chitinase in autolysis and cell death of Aspergillus nidulans. AB - Autolysis is a natural event that occurs in most filamentous fungi. Such self degradation of fungal cells becomes a predominant phenomenon in the absence of the regulator of G protein signaling FlbA in Aspergillus nidulans. Among a number of potential hydrolytic enzymes in the A. nidulans genome, the secreted endochitinase ChiB was shown to play a major role in autolysis. In this report, we investigate the roles of ChiB in fungal autolysis and cell death processes through genetic, biochemical, and cellular analyses using a set of critical mutants. Determination of mycelial mass revealed that, while the flbA deletion (DeltaflbA) mutant autolyzed completely after a 3-day incubation, the DeltaflbA DeltachiB double mutant escaped from hyphal disintegration. These results indicate that ChiB is necessary for the DeltaflbA-induced autolysis. However, importantly, both DeltaflbA and DeltaflbA DeltachiB strains displayed dramatically reduced cell viability compared to the wild type. These imply that ChiB is dispensable for cell death and that autolysis and cell death are separate processes. Liquid chromatography-tandem mass spectrometry analyses of the proteins that accumulate at high levels in the DeltaflbA and DeltaflbA DeltachiB mutants identify chitinase (ChiB), dipeptidyl peptidase V (DppV), O-glycosyl compound hydrolase, beta-N-acetylhexosaminidase (NagA), and myo-inositol-1 phosphate synthase (InoB). Functional characterization of these four genes reveals that the deletion of nagA results in reduced cell death. A working model bridging G protein signaling and players in autolysis/cell death is proposed. PMID- 19286988 TI - Nested genes CDA12 and CDA13 encode proteins associated with membrane trafficking in the ciliate Tetrahymena thermophila. AB - We describe a novel pair of nested genes, CDA12 and CDA13, from Tetrahymena thermophila. Both are implicated in membrane trafficking associated with cell division and conjugation. Green fluorescent protein localization reveals Cda12p decoration of diverse membrane-bound compartments, including mobile, subcortical tubulovesicular compartments; perinuclear vesicles; and candidates for recycling endosomes. Cda13p decorates intracellular foci located adjacent to cortically aligned mitochondria and their neighboring Golgi networks. The expression of antisense CDA12 RNA in transformants produces defects in cytokinesis, macronuclear segregation, and the processing of pinosomes to downstream compartments. Antisense CDA13 RNA expression produces a conjugation phenotype, resulting in the failure of mating pairs to separate, as well as failures in postconjugation cytokinesis and macronuclear fission. This study offers insight into the membrane trafficking events linking endosome and Golgi network activities, cytokinesis, and karyokinesis and the unique membrane-remodeling events that accompany conjugation in the ciliate T. thermophila. We also highlight an unusual aspect of genome organization in Tetrahymena, namely, the existence of nested, antisense genes. PMID- 19286989 TI - Are female healthcare workers at higher risk of occupational injury? AB - BACKGROUND: Differential risks of occupational injuries by gender have been examined across various industries. With the number of employees in healthcare rising and an overwhelming proportion of this workforce being female, it is important to address this issue in this growing sector. AIMS: To determine whether compensated work-related injuries among females are higher than their male colleagues in the British Columbia healthcare sector. METHODS: Incidents of occupational injury resulting in compensated days lost from work over a 1-year period for all healthcare workers were extracted from a standardized operational database and the numbers of productive hours were obtained from payroll data. Injuries were grouped into all injuries and musculoskeletal injuries (MSIs). Detailed analysis was conducted using Poisson regression modelling. RESULTS: A total of 42 332 employees were included in the study of whom 11% were male and 89% female. When adjusted for age, occupation, sub-sector, employment category, health region and facility, female workers had significantly higher risk of all injuries [rate ratio (95% CI) = 1.58 (1.24-2.01)] and MSIs [1.43 (1.11-1.85)] compared to their male colleagues. CONCLUSIONS: Occupational health and safety initiatives should be gender sensitive and developed accordingly. PMID- 19286990 TI - Continuing professional development in occupational medicine for general practitioners. AB - BACKGROUND: The Health and Occupation Reporting (THOR) network for general practitioners (GPs) offers free online continuing professional development (CPD) to contributing GPs. Use of this resource by members is relatively low. Non uptake is most frequently attributed to lack of time. AIMS: The primary aim was to assess the effects of changes made to educational material, available online to GPs participating in THOR-GP, 1 year after a needs assessment which informed the changes. The secondary aim, developed from the findings of the original study, was to compare contributing GPs who undertake work in occupational medicine, with those who do not, in terms of uptake of the educational material and of educational need. METHODS: GPs participating in THOR-GP, who responded to a questionnaire to assess their use of THOR-GP's website for CPD, were sent a follow-up questionnaire 1 year after the original survey. Both questionnaires comprised scales derived from the syllabus for the Diploma of the Faculty of Occupational Medicine and questions about attitudes to CPD in occupational medicine. RESULTS: No change was found in uptake of or rating of components of the website, following modification. Responders worked on average seven sessions per week in general practice and 1.5 sessions in occupational medicine. GPs working in occupational medicine reported greater confidence in some subject areas than GPs not currently working in occupational medicine and were also more likely to engage in CPD activity within the specialty of occupational medicine. CONCLUSIONS: Undertaking work in occupational medicine increases confidence in the subject and stimulates the use of related educational facilities. PMID- 19286991 TI - Pulmonary effects of spot welding in automobile assembly. AB - BACKGROUND: Spot welding is a type of resistance welding in which pieces of metals are pressed together and an electric current is passed through them. Spot welders are at risk of contact with some potentially hazardous agents but there are few studies about the respiratory effects of spot welding. AIMS: Our objective was to study lung function and respiratory symptoms among spot welders and office workers at an automobile assembly factory in Iran. METHODS: This was a cross-sectional study of 137 male spot welders and 129 office workers. We used a questionnaire to record demographic data, smoking habits, work history and respiratory symptoms. Spirometry was performed to assess lung function status. Metal fume samples from the respiratory zone of spot welders were analysed. RESULTS: The concentrations of metal fume were less than the American Conference of Industrial Hygienists (ACGIH) threshold limit values. There were significantly lower values for average forced expiratory volume in 1st second (FEV(1)), FEV(1)/forced vital capacity and 25-75% forced expiratory flow in spot welders compared to controls. There was also a significantly raised prevalence of respiratory symptoms (sputum and dyspnoea) in spot welders. Fifteen per cent of spot welders and 1% of controls had an obstructive pattern in spirometry. CONCLUSIONS: Our survey suggests that spot welders are at risk of developing respiratory symptoms and decreasing pulmonary function values despite their exposure to components of welding fume being within ACGIH guidelines. PMID- 19286992 TI - Occupational health advice in NICE guidelines. AB - BACKGROUND: Sickness absence is a major concern for society costing > pound 13 billion annually. Seeking medical advice is highly recommended in managing sickness absence. The National Institute for Health and Clinical Excellence (NICE) provides guidance on treating ill-health and promoting public health. NICE has published >500 guidelines. There are three versions of each guideline including full (comprehensive version), NICE (summary of recommendations) and public version. AIMS: To evaluate the availability of occupational health advice within relevant NICE guidelines. METHODS: Guidelines were selected on the basis of the impact of the disease and its treatment on work ability. All selected guidelines were reviewed for occupational health advice. The findings were categorized in five main domains: impact of the disease on work, impact of work on the disease, rehabilitation, fitness to work and modifications. RESULTS: Thirty-one published guidelines met the inclusion criteria. Advice on rehabilitation is most frequently referred to with 12 (39%) full, 8 (26%) NICE and 4 (13%) public information guidelines offering advice on this domain. Advice on return to work is the least frequent. The public versions of guidelines rarely offer advice on the studied domains. Occupational health organizations are enlisted as stakeholders in one guideline only. Two occupational health professionals have contributed to the development of two guidelines. CONCLUSIONS: This study suggests that occupational health professionals and organizations should participate more actively in developing NICE guidelines to integrate work related issues and occupational health advice into NICE guidelines. PMID- 19286993 TI - Occupational injuries and fatalities in copper mining in Zambia. AB - BACKGROUND: The metal mining industry employs approximately 15% of formally employed workers in Zambia, but there is little information about the magnitude of occupational injuries among the miners. AIMS: To determine the frequency rates of occupational injuries and fatalities among copper miners in Zambia. METHODS: A retrospective study of occupational injuries and fatalities at one of the largest copper mining companies in Zambia was undertaken for the period January 2005 to May 2007. Information on injuries and fatalities was obtained from the electronic accident survey database of the company. Analysis was restricted to fatalities and those injuries that had prompted medical attention and at least 1 day of absence from work. Annual injury and fatality frequency rates (injuries per 1000 employee years and fatalities per 100 000 employee years, respectively) were calculated. RESULTS: In the selected period, 165 injuries and 20 fatalities were recorded. The underground department had the highest frequency rates of fatalities (111/100 000 employee years) and injuries (5.5/1000 employee years). The most common cause of fatal injuries was fall of rock in the underground mines. The most frequent mechanism of injury was handling of tools and materials, and the most commonly injured body parts were the hands and fingers. CONCLUSIONS: The fatality rate is high compared to reported values from the metalliferous mining industry in developed countries, strongly suggesting that measures should be taken to reduce risks, particularly at underground sites. PMID- 19286994 TI - Musculoskeletal pain and night-shift naps in nursing home care workers. AB - BACKGROUND: Care workers in nursing homes are at high risk of developing musculoskeletal disorders (MSDs). Many care workers work in shifts, which may compromise both the quality of care they give and their working life. Taking a nap during night shifts has been proposed to ameliorate shift work-related problems, but its relationship with MSDs is not clear. AIMS: To explore how MSD pain differs according to frequency of night-shift naps. METHODS: A questionnaire study was conducted on 111 care workers at three nursing homes. Of 98 respondents, data from 66 shift workers (54 women) were analysed. Data on self rated pain in multiple sites (neck, shoulder, arm, leg and low back), naps during night shifts and relevant variables were collected. Participants were categorized into three groups on the basis of frequency of night-shift naps taken during the previous month: non-nappers, <50% nappers and >or =50% nappers. RESULTS: Pain at all sites, with the exception of low back pain, differed significantly among the three groups. Pain scores were lowest at the arm and leg for the > or =50% nappers. Neck and shoulder pain was lower for the > or =50% nappers and the non nappers compared to the <50% nappers. CONCLUSIONS: Reduced pain in the arm and leg was associated with taking a nap at least once every two night shifts among the nursing home care workers. No association was found between low back pain and night-shift naps in this sample. PMID- 19286995 TI - Influence of vibration exposure on tactile and thermal perception thresholds. AB - AIMS: To establish if intermittent exposure to hand-transmitted vibration had the same effect as continuous exposure on the temporary response of finger tactile and thermal perception thresholds. METHODS: Two laboratory experiments were conducted. In each, 10 healthy subjects, five males and five females, participated. The subjects' fingers were exposed to vibration under four conditions with a combination of different periods of exposure and rest periods. The vibration frequency was 125 Hz and the frequency-weighted acceleration was 5 m/s(2). A measure of the tactile or thermal perception was conducted before the different exposures to vibration. Immediately after the vibration exposure, the acute effect was measured continuously for the first 75 s. This was followed by regular measures for a maximum of 30 min. RESULTS: The results showed that combinations of vibration with different periods of exposure and rest periods significantly influenced vibrotactile perception, but not thermal perception. CONCLUSIONS: These findings suggest that intermittent exposure to hand transmitted vibration might be more beneficial for the response of the finger vibrotactile sensation than continuous exposure. This finding is inconsistent with the evaluation methods in ISO 5349-1 for vibrotactile sensation, but accurate for thermal perception. PMID- 19286996 TI - microRNA 184 regulates expression of NFAT1 in umbilical cord blood CD4+ T cells. AB - The reduced expression of nuclear factor of activated T cells-1 (NFAT1) protein in umbilical cord blood (UCB)-derived CD4+ T cells and the corresponding reduction in inflammatory cytokine secretion after stimulation in part underlies their phenotypic differences from adult blood (AB) CD4+ T cells. This muted response may contribute to the lower incidence and severity of high-grade acute graft-versus-host disease (aGVHD) exhibited by UCB grafts. Here we provide evidence that a specific microRNA, miR-184, inhibits NFAT1 protein expression elicited by UCB CD4+ T cells. Endogenous expression of miR-184 in UCB is 58.4 fold higher compared with AB CD4+ T cells, and miR-184 blocks production of NFAT1 protein through its complementary target sequence on the NFATc2 mRNA without transcript degradation. Furthermore, its negative effects on NFAT1 protein and downstream interleukin-2 (IL-2) transcription are reversed through antisense blocking in UCB and can be replicated via exogenous transfection of precursor miR 184 into AB CD4+ T cells. Our findings reveal a previously uncharacterized role for miR-184 in UCB CD4+ T cells and a novel function for microRNA in the early adaptive immune response. PMID- 19286997 TI - Efficient construction of producer cell lines for a SIN lentiviral vector for SCID-X1 gene therapy by concatemeric array transfection. AB - Retroviral vectors containing internal promoters, chromatin insulators, and self inactivating (SIN) long terminal repeats (LTRs) may have significantly reduced genotoxicity relative to the conventional retroviral vectors used in recent, otherwise successful clinical trials. Large-scale production of such vectors is problematic, however, as the introduction of SIN vectors into packaging cells cannot be accomplished with the traditional method of viral transduction. We have derived a set of packaging cell lines for HIV-based lentiviral vectors and developed a novel concatemeric array transfection technique for the introduction of SIN vector genomes devoid of enhancer and promoter sequences in the LTR. We used this method to derive a producer cell clone for a SIN lentiviral vector expressing green fluorescent protein, which when grown in a bioreactor generated more than 20 L of supernatant with titers above 10(7) transducing units (TU) per milliliter. Further refinement of our technique enabled the rapid generation of whole populations of stably transformed cells that produced similar titers. Finally, we describe the construction of an insulated, SIN lentiviral vector encoding the human interleukin 2 receptor common gamma chain (IL2RG) gene and the efficient derivation of cloned producer cells that generate supernatants with titers greater than 5 x 10(7) TU/mL and that are suitable for use in a clinical trial for X-linked severe combined immunodeficiency (SCID-X1). PMID- 19286998 TI - Targeted deletion of tumor suppressor PTEN augments neutrophil function and enhances host defense in neutropenia-associated pneumonia. AB - Neutropenia and related infections are the most important dose-limiting toxicities in anticancer chemotherapy and radiotherapy. In this study, we explored a new strategy for augmenting host defense in neutropenia-related pneumonia. Phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) signaling in neutrophils was elevated by depleting PTEN, a phosphatidylinositol 3' phosphatase that hydrolyzes PtdIns(3,4,5)P(3). In myeloid-specific PTEN knockout mice, significantly more neutrophils were recruited to the inflamed lungs during neutropenia-associated pneumonia. Using an adoptive transfer technique, we demonstrated that this enhancement could be caused directly by PTEN depletion in neutrophils. In addition, disruption of PTEN increased the recruitment of macrophages and elevated proinflammatory cytokines/chemokine levels in the inflamed lungs, which could also be responsible for the enhanced neutrophil recruitment. Depleting PTEN also significantly delayed apoptosis and enhanced the bacteria-killing capability of the recruited neutrophils. Finally, we provide direct evidence that enhancement of neutrophil function by elevating PtdIns(3,4,5)P(3) signaling can alleviate pneumonia-associated lung damage and decrease pneumonia-elicited mortality. Collectively, these results not only provide insight into the mechanism of action of PTEN and PtdIns(3,4,5)P(3) signaling pathway in modulating neutrophil function during lung infection and inflammation, but they also establish PTEN and related pathways as potential therapeutic targets for treating neutropenia-associated pneumonia. PMID- 19286999 TI - NPM-ALK inhibits the p53 tumor suppressor pathway in an MDM2 and JNK-dependent manner. AB - Anaplastic large cell lymphoma (ALCL) is characterized by the presence of the t(2;5)(p23;q35) generating the nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) fusion protein, a hyperactive kinase with transforming properties. Among these properties is the ability to regulate activity of the p53 tumor suppressor protein. In many human cancers, p53 is inactivated by mutation or other means, in some cases as a result of up-regulation of the negative regulator MDM2. However, the majority of ALK-expressing ALCL carry wild-type p53 and do not over express MDM2. We demonstrate a novel p53-dependent pathogenetic mechanism in ALK expressing lymphoma. We confirm previously published reports of NPM-ALK-induced activation of the phosphoinositide (PI) 3-kinase and Jun N-terminal kinase (JNK) stress-activated protein (SAP) kinase proteins, but in this study demonstrate a role for these in the regulation of p53 activity in an intricate signaling system. Specifically, constitutive ALK signaling leads to the functional inactivation and/or degradation of p53 in JNK and MDM2 dependent manners. We also show nuclear exclusion of p53 in a PI 3-kinase-dependent manner. Furthermore, we demonstrate that reactivation of p53 in ALK-expressing cells as a result of pharmacologic inhibition of JNK, PI 3-kinase, and/or MDM2 activities results in the induction of apoptosis suggesting a novel therapeutic modality. PMID- 19287000 TI - Multimorbidity in primary care: developing the research agenda. PMID- 19287001 TI - Minimally invasive surgical technique in total knee arthroplasty: a learning curve. AB - Clinical experience of learning a new technique of minimally surgery for total knee arthroplasty is presented. Close monitoring of the technique, pitfalls, learning tips, and tricks are discussed. A "learning phase" is identified as approximately 10 months or 21 knee replacements using minimally invasive technique. It took 50 operations before the surgical time equaled the open technique. There was no incidence of increased complications during the learning phase. Functional results such as stair climbing, walking distance, and walking with aids was significantly better after minimally invasive technique than after standard technique. PMID- 19287002 TI - CP12 from Chlamydomonas reinhardtii, a permanent specific "chaperone-like" protein of glyceraldehyde-3-phosphate dehydrogenase. AB - A new role is reported for CP12, a highly unfolded and flexible protein, mainly known for its redox function with A(4) glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Both reduced and oxidized CP12 can prevent the in vitro thermal inactivation and aggregation of GAPDH from Chlamydomonas reinhardtii. This mechanism is thus not redox-dependent. The protection is specific to CP12, because other proteins, such as bovine serum albumin, thioredoxin, and a general chaperone, Hsp33, do not fully prevent denaturation of GAPDH. Furthermore, CP12 acts as a specific chaperone, since it does not protect other proteins, such as catalase, alcohol dehydrogenase, or lysozyme. The interaction between CP12 and GAPDH is necessary to prevent the aggregation and inactivation, since the mutant C66S that does not form any complex with GAPDH cannot accomplish this protection. Unlike the C66S mutant, the C23S mutant that lacks the N-terminal bridge is partially able to protect and to slow down the inactivation and aggregation. Tryptic digestion coupled to mass spectrometry confirmed that the S-loop of GAPDH is the interaction site with CP12. Thus, CP12 not only has a redox function but also behaves as a specific "chaperone-like protein" for GAPDH, although a stable and not transitory interaction is observed. This new function of CP12 may explain why it is also present in complexes involving A(2)B(2) GAPDHs that possess a regulatory C-terminal extension (GapB subunit) and therefore do not require CP12 to be redox-regulated. PMID- 19287003 TI - A structural model for the damage-sensing complex in bacterial nucleotide excision repair. AB - Nucleotide excision repair is distinguished from other DNA repair pathways by its ability to process a wide range of structurally unrelated DNA lesions. In bacteria, damage recognition is achieved by the UvrA.UvrB ensemble. Here, we report the structure of the complex between the interaction domains of UvrA and UvrB. These domains are necessary and sufficient for full-length UvrA and UvrB to associate and thereby form the DNA damage-sensing complex of bacterial nucleotide excision repair. The crystal structure and accompanying biochemical analyses suggest a model for the complete damage-sensing complex. PMID- 19287004 TI - JAK2 and SHP2 reciprocally regulate tyrosine phosphorylation and stability of proapoptotic protein ASK1. AB - Previously we have shown that tyrosine 718 of ASK1 when phosphorylated is critical for SOCS1 binding and SOCS1-mediated degradation of ASK1. However, the kinase and phosphatase responsible for phosphorylation and dephosphorylation of ASK1 at Tyr-718 are unknown. In this study, we identified JAK2 and SHP2 as a Tyr 718-specific kinase and phosphatase, respectively. Interferon-gamma (IFN-gamma) induced degradation of ASK1 in normal but not in SOCS1-KO endothelial cells (EC). IFN-gamma-induced tyrosine phosphorylation of ASK1 at Tyr-718 was blocked by a JAK2-specific inhibitor. IFN-gamma enhanced the association between JAK2 and ASK1, and the ASK1-JAK2 complex was labile and was stabilized by the proteasomal inhibitor MG132. Furthermore, JAK2, but not JAK1, directly bound to and phosphorylated ASK1 at Tyr-718, leading to an enhanced association of ASK1 with SOCS1 and subsequent ASK1 degradation. Next, we showed that overexpression of the SH2-containing protein-tyrosine phosphatase-2 (SHP2) augmented, whereas a phosphatase-inactive mutant of SHP2 inhibited, TNF-induced ASK1 dephosphorylation. SHP2 associated with ASK1 in response to tumor necrosis factor in EC. An SHP-2 substrate-trapping mutant formed a complex with tyrosine phosphorylated ASK1, suggesting that ASK1 is a direct SHP2 substrate. Moreover, SHP2 wild type, but not a catalytically inactive mutant, dissociated SOCS1 from ASK1. IFN-gamma-induced ASK1 Tyr(P)-718 was enhanced in mouse EC deficient in SHP2 (SHP2-KO). In contrast, tumor necrosis factor-induced dephosphorylation of ASK1 at Tyr(P)-718 and activation of ASK1-JNK signaling, as well as EC apoptosis, are significantly reduced in SHP2-KO EC. Our data suggest that JAK2-SOCS1 and SHP2 reciprocally regulate ASK1 phosphorylation and stability in response to cytokines. PMID- 19287005 TI - Clathrin regulates the association of PIPKIgamma661 with the AP-2 adaptor beta2 appendage. AB - The AP-2 clathrin adaptor differs fundamentally from the related AP-1, AP-3, and AP-4 sorting complexes because membrane deposition does not depend directly on an Arf family GTPase. Instead phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) appears to act as the principal compartmental cue for AP-2 placement at the plasma membrane as well as for the docking of numerous other important clathrin coat components at the nascent bud site. This PtdIns(4,5)P(2) dependence makes type I phosphatidylinositol 4-phosphate 5-kinases (PIPKIs) lynchpin enzymes in the assembly of clathrin-coated structures at the cell surface. PIPKIgamma is the chief 5-kinase at nerve terminals, and here we show that the 26-amino acid, alternatively spliced C terminus of PIPKIgamma661 is an intrinsically unstructured polypeptide that binds directly to the sandwich subdomain of the AP-2 beta2 subunit appendage. An aromatic side chain-based, extended interaction motif that also includes the two bulky C-terminal residues of the short PIPKIgamma635 variant is necessary for beta2 appendage engagement. The clathrin heavy chain accesses the same contact surface on the AP-2 beta2 appendage, but because of additional clathrin binding sites located within the unstructured hinge segment of the beta2 subunit, clathrin binds the beta2 chain with a higher apparent affinity than PIPKIgamma661. A clathrin-regulated interaction with AP-2 could allow PIPKIgamma661 to be strategically positioned for regional PtdIns(4,5)P(2) generation during clathrin-coated vesicle assembly at the synapse. PMID- 19287006 TI - Structural basis of tRNA modification with CO2 fixation and methylation by wybutosine synthesizing enzyme TYW4. AB - Wybutosine (yW), one of the most complicated modified nucleosides, is found in the anticodon loop of eukaryotic phenylalanine tRNA. This hypermodified nucleoside ensures correct codon recognition by stabilizing codon-anticodon pairings during the decoding process in the ribosome. TYW4 is an S adenosylmethionine (SAM)-dependent enzyme that catalyzes the final step of yW biosynthesis, methylation and methoxycarbonylation. However, the structural basis for the catalytic mechanism by TYW4, and especially that for the methoxycarbonylation, have remained elusive. Here we report the apo and cofactor bound crystal structures of yeast TYW4. The structures revealed that the C terminal domain folds into a beta-propeller structure, forming part of the binding pocket for the target nucleoside. A comparison of the apo, SAM-bound, and S-adenosylhomocysteine-bound structures of TYW4 revealed a drastic structural change upon cofactor binding, which may sequester solvent from the catalytic site during the reaction and facilitate product release after the reaction. In conjunction with the functional analysis, our results suggest that TYW4 catalyzes both methylation and methoxycarbonylation at a single catalytic site, and in the latter reaction, the methoxycarbonyl group is formed through the fixation of carbon dioxide. PMID- 19287007 TI - The universal YrdC/Sua5 family is required for the formation of threonylcarbamoyladenosine in tRNA. AB - Threonylcarbamoyladenosine (t(6)A) is a universal modification found at position 37 of ANN decoding tRNAs, which imparts a unique structure to the anticodon loop enhancing its binding to ribosomes in vitro. Using a combination of bioinformatic, genetic, structural and biochemical approaches, the universal protein family YrdC/Sua5 (COG0009) was shown to be involved in the biosynthesis of this hypermodified base. Contradictory reports on the essentiality of both the yrdC wild-type gene of Escherichia coli and the SUA5 wild-type gene of Saccharomyces cerevisiae led us to reconstruct null alleles for both genes and prove that yrdC is essential in E. coli, whereas SUA5 is dispensable in yeast but results in severe growth phenotypes. Structural and biochemical analyses revealed that the E. coli YrdC protein binds ATP and preferentially binds RNA(Thr) lacking only the t(6)A modification. This work lays the foundation for elucidating the function of a protein family found in every sequenced genome to date and understanding the role of t(6)A in vivo. PMID- 19287008 TI - Padua University: the role it has played in the History of Medicine and Cardiology and its position today. PMID- 19287011 TI - Treatment of congenital cytomegalovirus infection: implications for future therapeutic strategies. AB - Cytomegalovirus (CMV) infection is the most common cause of congenital infection in the developed world, occurring in approximately 1% of all liveborns. Symptomatic disease occurs in 10% of all congenitally infected infants, resulting in a spectrum of clinical manifestations that include microcephaly, chorioretinitis, hepatosplenomegaly and sensorineural hearing loss, among others. Even those children who are asymptomatic at birth have a risk of hearing loss, with approximately 8% experiencing this sequela. Overall, congenital CMV infection accounts for one-third of all cases of sensorineural hearing loss. The economic burden of disease exceeds $2 billion annually in the USA. Therefore, this infection has been the target for antiviral therapy. Studies performed by the National Institute of Allergy and Infectious Diseases Collaborative Antiviral Study Group (CASG) have evaluated ganciclovir for the treatment of symptomatic congenital CMV infection with central nervous system involvement. In a randomized, controlled clinical trial of ganciclovir treatment (6 mg/kg iv every 12 h for 6 weeks) brainstem-evoked responses were utilized as the primary endpoint and demonstrated stabilization of hearing both at 6 months and >1 year. Treatment was associated with neutropenia in over 60% of treated patients. Since ganciclovir must be given intravenously, studies with its prodrug, valganciclovir, have been performed to assess pharmacokinetics and pharmacodynamics. Currently, a clinical trial of 6 weeks versus 6 months of valganciclovir is being performed by the CASG. Notably, only intravenous ganciclovir and orally administered valganciclovir have been used to treat congenital CMV infection. Hopefully, other drugs such as maribavir will be available for evaluation in this population. PMID- 19287012 TI - New model for dwelling dose calculation using Monte Carlo integration. AB - A new methodology and computer model using Monte Carlo simulation for indoor dose calculation are developed. A room model of six rectangular slabs of finite thickness with door or window in each slab was used. Point-kernel photon transport model with self-absorption correction was applied for dose calculations. New software was designed and programmed using Pascal programming language, which was evaluated for standard room design. The calculated dose due to natural radionuclides in the concert walls has differences from the average model results of 0.21% for (238)U, 12.3% for (232)Th and 13.9% for (40)K; and the variability of specific dose rate with changing position density and composition of walls was studied. The new model has more flexibility for real dose calculation of any room structure and tailing, which is not given in the published models. PMID- 19287013 TI - Evaluation of the entrance surface air kerma in mammographic examinations in Rio de Janeiro, Brazil. AB - The aim of this work was to evaluate the distribution of the entrance surface air kerma (ESAK) and the average glandular dose (D(G)) in four mammography facilities located in the city of Rio de Janeiro. The ESAK values were estimated from the X ray tube output rate (mGy/mAs) parameters. The image quality was evaluated by the radiologists in each clinic. The ESAK values obtained for a breast thickness of 45 mm were 5.58 mGy in Clinic A, 10.07 mGy in Clinic B, 13.89 mGy in Clinic C and 7.21 mGy in Clinic D. For D(G), it can be seen that, for the same compressed breast thickness (50 mm), the value varied from 0.20 to 3.60 mGy, with a mean value of 1.50 mGy for all the clinics. In image quality evaluation, Clinic D was the only one that presented a very low acceptability for quality criteria and inadequacies in relation to specks, masses and optical density. PMID- 19287014 TI - Reproductive biology of Datura wrightii: the benefits of a herbivorous pollinator. AB - BACKGROUND AND AIMS: A deeper understanding of mutualism can be reached by studying systems with measurable costs and benefits. Most studies of this type focus on an unusual class of obligate, species-specific pollination mutualisms. The interaction between Datura wrightii (Solanaceae) and the hawkmoth Manduca sexta offers similar advantages but greater generality. Adult moths both nectar at and deposit eggs on the same plant; larvae are herbivorous. The antagonistic component of this interaction has been well studied. Here the role of M. sexta as a pollinator of D. wrightii, particularly in the context of this moth's frequent nectaring visits to the bat-pollinated plant Agave palmeri, is documented. METHODS: Hand-pollinations were used to determine breeding system and the reproductive consequences of mixed loads of A. palmeri and D. wrightii pollen. Plants and moths were caged overnight to assess whether nectaring visits led to fruit and seed set. Finally, pollen deposited on field-collected stigmas was identified, with a particular focus on documenting the presence of D. wrightii and A. palmeri grains. KEY RESULTS: Datura wrightii is highly self-compatible, and a visit that deposits either outcross or self pollen almost doubles fruit and seed set compared with unvisited flowers. Manduca sexta transferred enough pollen to produce fruit and seed sets comparable to hand-pollination treatments. Agave palmeri did not interfere with D. wrightii success: in the field, stigmas received almost pure D. wrightii pollen, and hand-addition of large quantities of A. palmeri pollen had no measurable effect on fruit and seed set. CONCLUSIONS: The floral visitation component of the D. wrightii-M. sexta interaction is indeed mutualistic. This finding is essential background to future development of this interaction as a model system for studying mutualism's costs and benefits. It is already proving valuable for dissecting third-species effects on the outcome of mutualism. Results indicate that M. sexta's heavy visitation to A. palmeri has no negative effect on the benefits conferred to D. wrightii. However, it can be predicted to augment M. sexta populations to the point where the costs of the interaction begin to exceed its benefits. PMID- 19287015 TI - The quest for perfection: the contribution of the electrocardiogram to the prevention of sudden death in hypertrophic cardiomyopathy. PMID- 19287016 TI - Atrial fibrillatory rate and risk of stroke in atrial fibrillation. AB - AIMS: In atrial fibrillation (AF), a relation between electrocardiogram (ECG) parameters such as fibrillatory wave amplitude and stroke has been sought with conflicting results. In this study, we tested the hypothesis that the atrial fibrillatory rate of surface ECG lead V1 is related to stroke risk and may consequently be helpful for identifying high-risk patients. METHODS AND RESULTS: Atrial fibrillatory rate of 79 consecutive patients with AF and embolic stroke (age 83 +/- 7 years, 41% male) was compared with those of a matched AF population without stroke (n = 79). Atrial fibrillatory rate was determined from the surface ECG using spatiotemporal QRST cancellation and time-frequency analysis of lead V1. There was no significant difference in any clinical or echocardiographic variable in patients with stroke compared with AF controls without stroke. Atrial fibrillatory rate measured 373 +/- 55 fibrillations per minute (fpm; range 235 505 fpm) in the entire population. There was no fibrillatory rate difference between stroke patients (369 +/- 54 fpm, range 256-505 fpm) and AF controls without stroke (378 +/- 56 fpm, range 235-488 fpm). There was an inverse correlation between fibrillatory rate and age (R = -0.219, P = 0.006). Individuals aged >or=85 years had a significantly lower fibrillatory rate (356 +/ 44 fpm) than individuals aged 65-74 years (384 +/- 56 fpm, P = 0.033) and individuals aged 75-84 years (384 +/- 60 fpm, P = 0.016). In those subgroups, fibrillatory rates were, however, also similar in stroke patients and AF controls. CONCLUSION: Atrial fibrillatory rate obtained from surface ECG lead V1 is not a risk marker for stroke in AF. PMID- 19287017 TI - Long-term left ventricular reverse remodelling with cardiac resynchronization therapy: results from the CARE-HF trial. AB - AIMS: The aim of the present study was to assess the long-term effects of cardiac resynchronization therapy (CRT) on the reverse remodelling of the left ventricle (LV). METHODS AND RESULTS: The effects of CRT compared with controls on LV dimensions and function were assessed at 3, 9, and 18 months and at the end of study (average 29 months) in 735 (90%) patients with adequate echocardiographic examinations, randomized in the CARE-HF trial. Echocardiographic recordings were submitted to a core laboratory to ensure consistent quantitative analysis. LV volume decreased and ejection fraction increased substantially in the CRT group by 3 months and improved further at each assessment when compared with the control group. Effects were less marked in patients with ischaemic heart disease and those with right ventricular dysfunction, but not in patients with a restrictive LV filling pattern. The extent of reverse remodelling at 18 months showed a modest relationship with baseline interventricular mechanical delay (IVMD). CONCLUSION: CRT induces sustained LV reverse remodelling with the most marked effects occurring within the first 3-9 months. The extent of remodelling in response to CRT is related to the aetiology of heart failure and, to a lesser extent, to the IVMD. PMID- 19287019 TI - Response to: Trends in drug misuse recorded in primary care in the UK from 1998 to 2005. PMID- 19287020 TI - Women's health and gender-based clinical trials on etoricoxib: methodological gender bias. AB - BACKGROUND: The aim of this study was to determine compliance with published good practice guidelines for gender and clinical trials using etoricoxib. The rationale for choosing etoricoxib was that it is widely used by women and there is evidence of potential interaction with contraceptives and hormone replacement therapy as highlighted in the product characteristics. METHODS: The study reviewed 58 etoricoxib published trials (54 papers) to determine if they met the gender recommendations of the Guidelines of Food and Drug Administration (1993) and the Sex, Gender and Pain Special Interest Group Consensus Working Group Report (2007). RESULTS: Women formed 70% of a total of 49 835 subjects included in the etoricoxib trials, but only 31% of the subjects were in Phase I. About 85.7% of trials did not show sex-stratified data. About 90.6 and 93.3% did not provide efficacy and adverse effects data by sex, respectively. There is scarce information about the influence of issues that specifically affect women. Discussion Women are under-represented in the published etoricoxib trials, specifically, in Phase I. Sex-stratified data on efficacy and adverse effects are scarce in etoricoxib trials. Together with the lack of data on women-specific issues, this suggests that etoricoxib may pose the same potential problems for women as other cyclooxygenase-2 inhibitors. PMID- 19287021 TI - A to-do list for the new FDA commissioner. PMID- 19287022 TI - Rapid progression of submucosal invasive micropapillary carcinoma of the colon in progressive systemic sclerosis: report of a case. AB - A 71-year-old woman presented with hematochezia and narrowing of the stool. She suffered from progressive systemic sclerosis for 12 years and underwent home oxygen therapy due to pulmonary fibrosis and moderate pulmonary hypertension. Colonoscopy revealed a pedunculated, cauliflower-like polyp with a depressed surface in the sigmoid colon. The polyp was regarded as early colon cancer with possible submucosal invasion, and subsequent computed tomographic (CT) scans showed no evidence of lymph node involvement or distant metastases. Because of perioperative risks due to moderate pulmonary hypertension, she underwent an endoscopic resection of the early colon cancer. Pathological examination of the resected specimen of 20 mm diameter revealed the peculiar morphology of an adenocarcinoma with moderate lymphatic invasion. Immunohistochemical analysis for epithelial membrane antigen showed the specific 'inside-out growth pattern' indicative of invasive micropapillary carcinoma (IMPC). Taking the perioperative risks into consideration, she opted to undergo close follow-ups without an additional sigmoidectomy. At 6 months after the resection, the follow-up colonoscopy revealed a local recurrence of the colon cancer, and subsequent CT scans revealed multiple distant metastases including the lung, liver, lymph nodes and spleen. This is a rare case of a pure, submucosal IMPC of the colon. Furthermore, pure IMPC of the colon may represent a reliable predictor of lymphogenous and/or hematogenous metastases. Therefore, one should recommend an additional colectomy after endoscopic mucosal resection treatment when pathological findings confirmed IMPC of the colon and should continue a close follow-up for IMPC patients even when curative resections were performed at an early stage. PMID- 19287023 TI - A phase 2 clinical trial of panitumumab monotherapy in Japanese patients with metastatic colorectal cancer. AB - OBJECTIVE: Panitumumab, a fully human monoclonal antibody targeting epidermal growth factor receptor (EGFR), has antitumor activity and an acceptable safety profile in patients with metastatic colorectal cancer (mCRC). This Phase 2 study evaluated efficacy, pharmacokinetics and safety of panitumumab in Japanese patients with mCRC who developed progressive disease during or after fluoropyrimidine, irinotecan and oxaliplatin chemotherapy. METHODS: Eligible patients had histologically proven colorectal adenocarcinoma and EGFR tumor expression in > or =1% of tumor cells by immunohistochemistry. Patients received panitumumab 6 mg/kg every 2 weeks until disease progression or unacceptable toxicity. The primary endpoint was objective response rate (ORR) per modified Response Evaluation Criteria in Solid Tumors (RECIST) by independent central review. Secondary endpoints included progression-free survival (PFS), overall survival (OS), pharmacokinetic parameters and incidence of adverse events. RESULTS: Fifty-two patients received at least one dose of panitumumab. Seven patients had partial responses for a confirmed ORR of 13.5% (95% CI: 5.6, 25.8). Median PFS was 8.0 weeks (95% CI: 7.4, 11.4) and median OS was 9.3 months (95% CI: 7.1, 12.8). Panitumumab pharmacokinetics were consistent with prior studies in Japanese and non-Japanese patients. The most common treatment-related adverse events (all, worst grade 3) were acne (81%, 2%), dry skin (62%, 0%), rash (46%, 2%), paronychia (33%, 2%), pruritus (33%, 0%) and hypomagnesemia (33%, 0%). No adverse event of infusion reaction was reported by the investigators. CONCLUSIONS: Panitumumab monotherapy was active in Japanese patients with chemotherapy-refractory mCRC, with pharmacokinetic and safety profiles similar to those seen in prior studies. PMID- 19287024 TI - Recognition of non-self-polysaccharides by C-type lectin receptors dectin-1 and dectin-2. AB - The discovery of several transmembrane receptors expressed by antigen presenting cells, including those that detect and interact with specific sugar moieties on the surface of microbes, has improved our understanding of how immunity against infection is generated. This knowledge, in turn, prompted us to review such interactions with emphasis on C-type lectin receptors and a focus on the roles of dectin-1 and dectin-2 in anti-fungal immunity. PMID- 19287025 TI - Action potential initiation in unmyelinated afferents comes in from the cold. PMID- 19287026 TI - A self-activating intrinsic brake on bursting in CA3 neurons. PMID- 19287027 TI - Effect of muscle injection site on preanaesthetic sedation in dogs. AB - The demeanour of 122 dogs was assessed, and a mixture of 0.025 mg/kg acepromazine and 0.3 mg/kg morphine was injected intramuscularly into one of four randomly assigned muscle sites (cervical epaxial, triceps brachii, middle gluteal and quadriceps femoris) and the dogs' reactions to the injections were assessed. The development of sedation and the occurrence of side effects were assessed after 10, 20 and 30 minutes, and each dog's reaction to venous catheterisation was scored. All the dogs became similarly sedated after 30 minutes. The degree of sedation and the incidence of side effects were independent of the injection site, but the dogs receiving intragluteal injections had lower sedation scores after 10 minutes. The dogs' demeanour had no effect on their response to venous catheterisation. The dogs receiving an injection into the postural quadriceps and triceps muscles showed more marked reactions than those injected into the non postural cervical epaxial and gluteal muscles. PMID- 19287028 TI - Neurological signs in juvenile harbour seals (Phoca vitulina) with fatal phocine distemper. AB - In 2002, the northern European harbour seal (Phoca vitulina) population experienced an epidemic of phocine distemper virus (PDV) in which 22,000 seals died. Clinical signs were recorded in 20 harbour seal pups admitted to the Seal Rehabilitation and Research Centre with clinical disease, and they were diagnosed PDV infection-positive by RT-PCR postmortem. All 20 had respiratory signs, 14 had conjunctivitis and 10 had neurological signs. Severe neurological signs were one of the criteria for euthanasia during the epidemic, and many pups that were euthanased were not included in this study owing to the lack of complete datasets. Neurological signs were therefore among the most prevalent signs of fatal PDV infection in harbour seal pups. The lymphoid depletion reported in dead seals during the epidemic was not reflected in the total mononuclear leucocyte count of the seal pups, but they had an absolute granulocytosis, thrombocytosis, anaemia, and high total white blood cell counts. When first examined, 11 of the pups had a positive serum IgG titre, and four had a positive serum IgM titre. High levels of PDV-specific serum IgG antibodies were not correlated with an absence of clinical signs or longer survival. PMID- 19287029 TI - Hypospadias in six dogs. AB - Six dogs, five males and one hermaphrodite, were diagnosed with hypospadias. Two of the five males had the penile form of the condition, two had the perineal form and one had the glandular form; the hermaphrodite had the scrotal form. The hermaphrodite had no prostate gland and no right testicle; it had a normal right ovary and horn of the uterus but the left horn was joined to the testicle. Its karyotype showed 78 chromosomes, all in metaphase, and two typical sex chromosomes X and Y. PMID- 19287030 TI - Time course of plumage damage in commercial layers. PMID- 19287031 TI - Clinical and laboratory findings in a Eurasian badger (Meles meles) with pituitary dwarfism. PMID- 19287032 TI - Molecular characterisation of two strains of Anaplasma platys in Brazil. PMID- 19287033 TI - Doppler-derived evidence of severe pulmonary arterial hypertension in a pony with recurrent airway obstruction. PMID- 19287034 TI - Fatal candidiasis in a wild red squirrel (Sciurus vulgaris). PMID- 19287035 TI - Rubber bands as a cause of lameness in cattle. PMID- 19287036 TI - Cutaneous mast cell tumours in dogs. PMID- 19287037 TI - Mastitis control plan. PMID- 19287038 TI - Upside to the downturn? PMID- 19287039 TI - What is veterinary forensics? PMID- 19287040 TI - Reindeer health and welfare. PMID- 19287041 TI - RBP4 variants are significantly associated with plasma RBP4 levels and hypertriglyceridemia risk in Chinese Hans. AB - We previously found that plasma RBP4 levels were strongly associated with metabolic syndrome components. This study aimed to determine whether RBP4 variants are associated with the metabolic syndrome components and plasma RBP4 levels, and to investigate whether the associations between plasma RBP4 and the metabolic syndrome components are causal. Five tagSNPs were tested for their associations with plasma RBP4 levels and metabolic syndrome components in a population-based sample of 3,210 Chinese Hans. A possible causal relationship between plasma RBP4 levels and hypertriglyceridemia was explored by Mendelian randomization. Plasma RBP4 levels were significantly associated with rs10882273 (betaz -0.10SD[-0.17, -0.03], P = 0.0050), rs3758538 (betaz -0.13SD[-0.24, 0.02], P = 0.0249) in all participants, and with rs17108993 in Shanghai participants (betaz -0.19SD[-0.32, -0.05], P = 0.0061). The single nucleotide polymorphism (SNP) rs3758538 was significantly associated with hypertriglyceridemia (OR 0.62[0.45-0.85], P = 0.0026) and triglycerides (betaz 0.19SD[-0.30, -0.07], P = 0.001) in all participants. In Mendelian randomization analysis, the observed effect size of association between rs3758538 and hypertriglyceridemia was different from the expected effect size (P = 0.0213). This is the first study to show that the RBP4 variants are significantly associated with plasma RBP4 levels and hypertriglyceridemia risk in Chinese Hans. However, results of Mendelian randomization do not support the hypothesis that RBP4 levels are causally related to hypertriglyceridemia risk. PMID- 19287042 TI - Reduced insulin-mediated inhibition of VLDL secretion upon pharmacological activation of the liver X receptor in mice. AB - The nuclear liver X receptor (LXR) regulates multiple aspects of cholesterol, triacylglycerol (TG), and carbohydrate metabolism. Activation of LXR induces the expression of genes encoding enzymes involved in de novo lipogenesis (DNL) resulting in hepatic steatosis in mice. Pharmacological LXR activation has also been reported to improve insulin sensitivity and glucose homeostasis in diabetic rodents. The effects of pharmacological LXR ligands on insulin's action on hepatic lipid metabolism are not known. We evaluated secretion of VLDL during a hyperinsulinemic euglycemic clamp in mice treated with the LXR-ligand T0901317. In untreated mice, hyperinsulinemia reduced the availability of plasma NEFA for VLDL-TG synthesis, increased the contribution of DNL to VLDL-TG, reduced VLDL particle size, and suppressed overall VLDL-TG production rate by approximately 50%. Upon T0901317 treatment, hyperinsulinemia failed to reduce VLDL particle size or suppress VLDL-TG production rate, but the contribution of DNL to VLDL-TG was increased. In conclusion, the effects of LXR activation by T0901317 on lipid metabolism can override the normal control of insulin to suppress VLDL particle secretion. PMID- 19287043 TI - Period correction of the QTc of moxifloxacin with multiple predose baseline ECGs is the least variable of 4 methods tested. AB - This study compares 4 baseline correction methods on the effect of moxifloxacin on the QT/QTc interval: (1) day -1 time-matched baseline electrocardiograms (ECGs), (2) 3 triplicate predose ECGs, (3) 1 triplicate predose ECG, and (4) no baseline correction. Forty-four healthy subjects receive a single dose of moxifloxacin (400 mg), placebo, and 2 doses of an investigational agent in a 4 period crossover fashion. For all 4 methods, the largest mean difference from placebo in the moxifloxacin study-specific QTc is 11.97 to 13.23 ms and occurs at 3 to 4 hours postdose; the lower 90% confidence interval is greater than 5 ms from 2 to 8 hours. The average standard error of the mean is 1.36 ms for 3 triplicate predose ECGs, 1.40 ms for 1 triplicate predose ECG, 1.60 ms for day -1 time-matched baseline ECGs, and 1.65 ms for no baseline correction. Predose baseline methods (3 or 1 triplicate ECGs) are superior to the day -1 time-matched baseline correction or without baseline correction. PMID- 19287044 TI - Dysfunction of the social brain in schizophrenia is modulated by intention type: an fMRI study. AB - In this fMRI study, we investigated theory of mind (ToM) in patients with paranoid schizophrenia. We hypothesized that the network supporting the representation of intentions is dysfunctional in patients with schizophrenia dependent on the type of intention involved. We used a paradigm including a control condition (physical causation) and three intention conditions (private intention, prospective social intention and communicative intentions) differing in the degree of social interaction. In all four experimental conditions patients performed worse than controls regarding accuracy and reaction time. They showed significantly less activation in three regions typically activated in ToM tasks, i.e. paracingulate cortex and bilateral temporo-parietal junctions. However, this dysfunction was dependent on the type of intention represented, i.e. was present only for social but not for non-social intentions. Moreover, part of the reduced activation was related to the fact that there was no signal drop in these regions for the physical causality condition as usually found in controls. This may be due to the tendency of schizophrenic patients to attribute intentionality to physical objects. Our findings have implications for the study and understanding of ToM in schizophrenia but also in other disorders like autism. PMID- 19287045 TI - Doses to patients from photoneutrons emitted in a medical linear accelerator. AB - This study of doses to patients from emitted photoneutrons in a medical linear accelerator (Varian 2100C) was carried out. Dose calculation was performed using Monte Carlo Geant4 code. The model was used to calculate the neutron fluence, as a function of the neutron energy inside the treatment room to estimate the effective dose to patients. The ambient dose equivalent versus field size for patients is reported in this study. The ambient dose equivalent using 1 x 1 cm(2) field size, at isocentre and X-ray modes of 20, 18, 15 and 10 MV, was found to be 1.85, 1.79, 0.61 and 0.06 mSv Gy(-1), respectively. The mean energies of emitted photoneutrons were 0.48, 0.44, 0.40 and 0.16 MeV at X-ray modes of 20, 18, 15 and 10 MV, respectively. The results of ambient dose equivalent from emitted photoneutrons cannot be ignored and can represent a risk for healthy tissues. This study emphasised that Geant4 Monte Carlo code is an appropriate choice for studying photoneutron production and transport. PMID- 19287046 TI - RCT of real versus placebo acupuncture in IVF. PMID- 19287047 TI - The longitudinal impact of partner coping in couples following 5 years of unsuccessful fertility treatments. AB - BACKGROUND: Because there is a lack of longitudinal research examining the impact of partner coping in couples experiencing infertility, we know very little about the long-term nature of coping with infertility and how partner coping strategies impact personal, marital and social distress. METHODS: Participants were Danish men and women about to start a cycle of assisted reproduction treatment who were followed for a 5 year period of unsuccessful treatments. Multilevel modeling using the actor-partner interdependence model was used to examine the couple as the unit of analysis. RESULTS: Active and passive avoidance coping strategies were significantly related to increased personal, marital and social distress at the individual and partner level. Meaning-based coping strategies were related to decreases in a woman's individual distress and her partner's marital distress. CONCLUSIONS: Partner coping strategies have a significant impact on the other member of the couple over time in men and women undergoing infertility treatments over a 5 year period. Physicians and mental health professionals can educate men and women regarding the ineffectiveness of avoidance coping strategies as well as the beneficial nature of finding new meaning and life goals while experiencing the stress of infertility. PMID- 19287048 TI - Regulation of vascular physiology and pathology by the S1P2 receptor subtype. AB - Sphingosine-1-phosphate (S1P) is now recognized as a lipid mediator that acts via G-protein-coupled receptors. S1P receptors couple to various heterotrimeric G proteins and regulate downstream targets and ultimately cell behaviour. The prototypical S1P1 receptor is known to couple to Gi and regulates angiogenesis, vascular development, and immune cell trafficking. In this review, we focus our attention on the S1P2 receptor, which has a unique G-protein-coupling property in that it preferentially activates the G(12/13) pathway. Recent studies indicate that the S1P2 receptor regulates critical intracellular signalling pathways, such as Rho GTPase, the phosphatase PTEN, and VE-cadherin-based adherens junctions. Analysis of mutant mice has revealed the critical role of this receptor in inner ear physiology, heart and vascular development, vascular remodelling, and vascular tone, permeability, and angiogenesis in vertebrates. These studies suggest that selective modulation of S1P2 receptor function by pharmacological tools may be useful in a variety of pathological conditions. PMID- 19287049 TI - Flux balance analysis of biological systems: applications and challenges. AB - Systems level modelling and simulations of biological processes are proving to be invaluable in obtaining a quantitative and dynamic perspective of various aspects of cellular function. In particular, constraint-based analyses of metabolic networks have gained considerable popularity for simulating cellular metabolism, of which flux balance analysis (FBA), is most widely used. Unlike mechanistic simulations that depend on accurate kinetic data, which are scarcely available, FBA is based on the principle of conservation of mass in a network, which utilizes the stoichiometric matrix and a biologically relevant objective function to identify optimal reaction flux distributions. FBA has been used to analyse genome-scale reconstructions of several organisms; it has also been used to analyse the effect of perturbations, such as gene deletions or drug inhibitions in silico. This article reviews the usefulness of FBA as a tool for gaining biological insights, advances in methodology enabling integration of regulatory information and thermodynamic constraints, and finally addresses the challenges that lie ahead. Various use scenarios and biological insights obtained from FBA, and applications in fields such metabolic engineering and drug target identification, are also discussed. Genome-scale constraint-based models have an immense potential for building and testing hypotheses, as well as to guide experimentation. PMID- 19287050 TI - Shift to Th2 cytokine response in dengue haemorrhagic fever. PMID- 19287051 TI - Ectopic Cushing's syndrome: some facts. PMID- 19287052 TI - Challenges in providing HIV care to paediatric age group in India. PMID- 19287053 TI - Animal models of tuberculosis for vaccine development. AB - Animal models for testing different vaccine candidates have been developed since a long time for studying tuberculosis. Mice, guinea pigs and rabbits are animals most frequently used. Each model has its own merits for studying human tuberculosis, and none completely mimics the human disease. Different animal models are being used depending upon the availability of the space, trained manpower as well as other resources. Efforts should continue to develop a vaccine which can replace/outperform the presently available vaccine BCG. PMID- 19287054 TI - Informed consent & ethical issues in paediatric psychopharmacology. AB - Issues relating to informed consent and ethics in paediatric psychopharmacology limit research in this population. Children vary in their levels of cognitive development, and presence of psychiatric disorder may further impair their ability to give informed consent. In decisional impairment subjects, various methods used for consent are assent/dissent; inclusion of advance directives; and/or alternative decision-makers. India is emerging as a new market for clinical trials in recent years. Moreover, in India the sociocultural realities are different from those in the western countries making it necessary for professionals to be cautious in conducting drug trials. In this review, issues regarding informed consent in children and adolescent with psychiatric diagnosis are discussed for information, discussion and debate by professionals, parents, society and legal experts to create awareness and to facilitate development of guidelines that are appropriate and applicable to the Indian system. PMID- 19287055 TI - Ectopic Cushing's syndrome: experience from a tertiary care centre. AB - BACKGROUND & OBJECTIVE: Ectopic secretion of adrenocorticotropic hormone (ACTH) is rare, contributing to 10 per cent cases of endogenous Cushing's syndrome. We describe our experience of about two decades of patients with ectopic Cushing's syndrome (ECS) seen at a tertiary care centre from north India. METHODS: Records of patients with ECS from 1985 to 2006 were retrospectively reviewed that included the presenting manifestations, clinical symptoms and signs, biochemical investigations including plasma cortisol, ACTH and high dose dexamethasone suppression test (HDDST), imaging modalities to localize the non pituitary source of ACTH production, management and follow up of these patients. RESULTS: The study group included 12 patients (7 men) with mean (+/-SD) age at presentation 27.6 +/- 9.5 yr (range 13 to 48 yr) and the mean lag period between onset of symptoms and the diagnosis was 18.3 +/- 12.9 months with a range of 3 to 48 months. The weight loss (41.7%) followed by hyperpigmentation (25%) and infections (16.7%) were the common presenting manifestations. Cuticular atrophy (100%), hypertension (100%), bruise (92%) and proximal myopathy (83%) were the commonest signs. Plasma cortisol at 0800 h was 1267.3 +/- 483.3 nmol/l and at 2200 h was 1214.9 +/- 442.6 nmol/l indicating loss of circadian rhythm. The mean plasma ACTH was 221.1 +/- 55.9 (range 21.7 to 950 pg/ml). All but 2 patients had non-suppressibility of 0800 h plasma cortisol with HDDST. Five patients had thymic carcinoid, 3 had bronchial and one each had islet, hepatic and gut carcinoid and one had medullary thyroid carcinoma as a cause of ectopic ACTH secretion. The mean duration of follow up of these patients was 42.6 months and only two could sustain cure while remaining had either residual or recurrence. INTERPRETATION & CONCLUSION: Ectopic Cushing's syndrome is a rare disease with varied manifestations and associated with increased morbidity and mortality. It presents with clinical features quite similar to classical Cushing's. Surgery with removal of primary tumour was found to be treatment of choice. PMID- 19287056 TI - Demographic & clinical profile of HIV infected children accessing care at Tambaram, Chennai, India. AB - BACKGROUND & OBJECTIVE: Human immunodeficiency virus (HIV) is severely affecting the poorly educated and economically disadvantaged in Indian society. When children start developing clinical manifestations, needing treatment, they have to travel long distances for accessing care and support at tertiary institutions. This places an extra burden on patients, who are already struggling to cope with their illness. Sufficient data are needed for the government to evolve appropriate policy for providing care to the children affected with HIV. We undertook this study to present the socio-demographic characteristics, signs and symptoms, clinical profile, distance travelled and follow up pattern of HIV positive children who accessed care for the first time in a referral hospital at Chennai, India. METHODS: Electronic medical records from patients diagnosed with HIV between 2002 and 2004 at the Government Hospital for Thoracic Medicine (GHTM) in Tambaram (Chennai) in India were analyzed to understand care-seeking behaviours. Demographic variables such as age, sex, education and occupation, data on clinical manifestations were examined together with geographic information. RESULTS: At GHTM 1,768 new paediatric patients accessed care from 2002 to 2004. Children aged less than 5 yr were 49.9 per cent; 1115 children had (63%) tuberculosis. Significantly, 14.9 and 20.6 per cent children had extra pulmonary TB and disseminated TB respectively. Lower respiratory infection (15.8%), Pneumocystis carinii pneumonia (15.20%), oral/oesophageal candidiasis (13.5%), wasting (6.1%) and diarrhoeal disorders (3.5%) were the common clinical manifestations. In all 47 per cent children traveled between 200-400 km from home and 14 per cent travelled over 400 km. INTERPRETATION & CONCLUSION: Our findings showed that tuberculosis should be regarded as the indicator disease for HIV infection in children, especially when they have clinical manifestations of progressive, non pulmonary and disseminated disease. The primary and secondary health care centres should have the trained capacity to diagnose and treat HIV disease and opportunistic infections so as the children to have much needed care and support nearer to their residence. PMID- 19287057 TI - Improved estimates of India's HIV burden in 2006. AB - BACKGROUND & OBJECTIVE: HIV estimates in India were based on HIV sentinel surveillance (HSS) data and several assumptions. Expansion of sentinel surveillance to all districts and community based HIV prevalence measured by National Family Health Survey-3 (NFHS-3) in 2006 provided opportunity to replace many of the assumptions with evidence based information and improve the HIV estimate closer to reality. This article presents a detailed account of the methodology used for the 2006 HIV burden estimates for India. METHODS: State-wise adult HIV prevalence among different risk groups observed from HSS 2006 was adjusted for site level variations using a random effects model and for the previous four years the same was back calculated using trend equations derived from a mixed effects logistic regression model based on consistent sites prevalence. The adjusted HIV prevalence among the general population was calibrated to the estimates from NFHS-3. Overall point estimates of adult HIV prevalence in each State for 2002-2006 were derived from the UNAIDS Workbook and projected for the period 1985-2010. The results were put into Spectrum to derive estimates of the number of people living with HIV in all ages and other epidemic impacts. RESULTS: National adult HIV prevalence was 0.36 per cent (range 0.29 0.46%) and the estimated number of people living with HIV was 2.47 million (range 2.0-3.1 million) in 2006. The national adult HIV prevalence remains stable around 0.4 per cent between 2002 and 2006. The States with the highest estimated prevalence were Manipur, Nagaland and Andhra Pradesh. The States with the highest burden were Andhra Pradesh, Maharashtra, Karnataka and Tamil Nadu. INTERPRETATION & CONCLUSION: The improvement in the 2006 estimates of the HIV burden in India is attributable to the expanded sentinel surveillance and representative data from the population-based survey in 2006, combined with an improved analysis. Despite the downward revision, India continues to face a formidable challenge to provide prevention, treatment and care to those in need. PMID- 19287058 TI - Establishment of T-lymphocyte subset reference intervals in a healthy adult population in Chennai, India. AB - Estimation of CD4+ T-lymphocytes continues to be an important aspect for monitoring HIV disease progression and response to antiretroviral therapy. Most of the diagnostic laboratories often rely on western text books for CD4+ T lymphocyte reference values, which could, often be unreliable for usage in local settings. Therefore, we attempted to establish the reference values for T lymphocyte subsets among healthy adults in a cross-sectional study carried out at the YRG Centre for AIDS Research and Education (YRG CARE) in Chennai, south India, in 213 (84 female and 129 male) healthy, HIV-1/2 seronegative adults as volunteers. Whole blood specimens were processed for CD4+, CD8+ T-lymphocyte estimation and haematological parameters. The established range of CD4+ T lymphocyte counts for men and women were 383-1347 cells/microl (mean 865 and median 845 cells/microl) and 448-1593 cells/microl (mean 1021 and median 954 cells/microl), respectively. Women had significantly higher absolute CD4+ Tlymphocyte counts (P<0.001) and CD4+:CD8+ T-lymphocyte ratio as compared to men. The established normal range of CD4+ T-lymphocyte % was 21-59 (mean 40.2 and median 40.1). The influence of age was not observed in any of the parameters except CD4+/CD8+ T-lymphocyte ratio with the >45 yr age group. Further studies with greater sample size may be required to define the staging of HIV disease in relation to the normal CD4 T-lymphocyte count in the general population. PMID- 19287059 TI - Role of acute viral hepatitis as a confounding factor in antituberculosis treatment induced hepatotoxicity. AB - BACKGROUND & OBJECTIVE: Drug induced hepatotoxicity (DIH) is an important and commonly encountered adverse effect with antituberculosis (anti-TB) treatment. Acute viral hepatitis (AVH) is an important confounding reason which clinically, biochemically and histologically mimics DIH. METHODS: The contributory role of acute viral hepatitis as a confounding factor in patients with normal baseline liver functions who developed acute hepatitis while receiving short-course anti TB treatment was prospectively studied. The sera of all patients who developed acute hepatitis were analysed for markers for hepatitis A, B, C and E viruses. RESULTS: Viral hepatitis was present in 15 of the 102 (14.7%) patients who developed acute hepatitis while receiving anti-TB treatment with hepatitis E virus being the most common cause Later onset of acute hepatitis [58 (5-133) vs. 26 (3-221) days; P=0.04], large elevations in aspartate aminotransferase (AST) [371 (30-2643) vs. 212 (63-1990 IU/l); P=0.03] and alanine aminotransferase (ALT) [388 (31-2997) vs. 225 (52- 1670 IU/l); P= 0.002] and a longer time for normalization of deranged liver functions [36.7 +/- 13.3 vs. 24.5 +/- 19.3 days; P=0.02] indicated acute viral hepatitis as the cause of liver function derangement. INTERPRETATION & CONCLUSION: Our findings showed AVH in 14.7 per cent patients who developed hepatotoxicity while an anti-TB treatment. Therefore, in endemic areas, viral hepatitis should be sought after and excluded in all patients suspected to have DIH before attributing the hepatotoxic effect to the anti-TB drugs. PMID- 19287060 TI - Endothelial nitric oxide synthase (eNOS) Glu298-->Asp polymorphism (G894T) among south Indians. AB - BACKGROUND & OBJECTIVE: Nitric oxide (NO) synthesized by endothelial cells is known to be a potent vasodilator. It has been suggested that the single nucleotide polymorphism (SNP) in endothelial nitric oxide synthase (eNOS) (G894T) may affect the response of vascular endothelium to increased oxidative stress. The Glu298Asp (GT) polymorphism located in exon 7, may be associated with several diseases. The objective of the present study was to determine the presence of genotype frequencies of Glu298-->Asp (G894T) single nucleotide polymorphism in the eNOS gene among south Indian male Tamil speaking population. METHODS: Polymerase chain reaction and restriction fragmant analysis was done to detect the presence of Glu298-->Asp (G894T) variant of the eNOS gene in 105 healthy male volunteers. RESULTS: The frequency of the eNOS GG, GT, and TT genotypes was found to be 74.3, 25.7, and 0 per cent respectively. The 'T' allele frequency among this population was found to be 0.13. Chi square analysis showed that the study population lies in accordance with Hardy-Weinberg equilibrium. INTERPRETATION & CONCLUSION: The occurrence of the homozygous mutant (TT) was completely absent among the study population. Moreover, our result indicated that the presence of the homozygous mutant genotype for eNOS polymorphism was absent/less common among south Indian male Tamil population of Asian race. PMID- 19287061 TI - Murine monoclonal anti-H as typing reagent. AB - BACKGROUND & OBJECTIVE: The antigen H present on the surface of red cells in varying concentration, is maximum in O group red cells, but absent in Bombay phenotype individuals. This differentiation is generally detected by seed extracts of Ulex europaeus. The titre of such an extract is usually low and is subjected to batch variation. Hence, we carried out this study to raise potent murine monoclonal antibody against H antigen. METHODS: Spleen cells of female BALB/c mice immunized with O group red cells were fused in presence of polyethylene glycol (PEG) 1500 with a mouse myeloma cell line Sp2/0 Ag14 in hypoxanthine aminopterine thymidine (HAT) selective medium and incubated at 37 degrees, 5 per cent CO(2) and 95 per cent humidity for a week. RESULTS: The culture supernatants showing anti-H activity, were further subcloned and two clones 3E8A10 and 3E8A11 generated which showed a good potency, avidity and specificity. INTERPRETATION & CONCLUSION: The anti-H clones thus produced indigenously provided a potent reagent in distinguishing normal O group from Bombay phenotype individuals. The unlimited availability makes this reagent cost effective to ensure a constant supply of hybrid clones with the similar specificities. PMID- 19287062 TI - Systemic manifestations in chronic arsenic toxicity in absence of skin lesions in West Bengal. AB - BACKGROUND & OBJECTIVE: Pigmentation and keratosis are the prerequisites to diagnose arsenicosis. However, many systemic manifestations occur in association with pigmentation and keratosis in people exposed to chronic drinking of arsenic contaminated water. The present study aim to find out whether systemic manifestations occur in significant number of cases in arsenic exposed people in the absence of skin lesions in an affected district in West Bengal, India. METHODS: A cross-sectional study was carried out in South 24 Parganas, an arsenic affected district of West Bengal, India. Both dermatological and systemic manifestations were recorded and water samples collected for arsenic analysis from 7683 participants. A correlation of systemic manifestations in relation to arsenic exposure was carried out in subjects having no arsenical skin lesion. Prevalence odds ratio (POR) was calculated for each outcome comparing those with high arsenic exposure with those with lowest exposure. RESULTS: The frequency of occurrence of various clinical manifestations like weakness, anaemia, diarrhoea, hepatomegaly and lung disease was found to be significantly higher among participants drinking water having arsenic concentration > or = 50 microg/l in comparison to those taking water with arsenic content below this level. Further, there was increased occurrence of these manifestations with increasing concentration of arsenic level in drinking water, and this followed a dose response relationship. INTERPRETATION & CONCLUSION: It appears that it is worthwhile to include people with systemic manifestations in absence of skin lesions with evidence of arsenic exposure as suspected cases of arsenicosis for case detection and in surveillance programme. PMID- 19287063 TI - An exploratory clinical study of adult attention deficit/hyperactivity disorder from India. AB - BACKGROUND & OBJECTIVE: Although adult attention-deficit/hyperactivity disorder (ADHD) is well recognized in western countries, there are no reports of adult ADHD from India. Therefore, this study was planned to identify ADHD in adults in an Indian psychiatry outpatient setting and elicit its phenomenology and co morbidities. METHODS: A total of 283 adults were screened using Adult ADHD Self Report Scale-v1.1 (ASRSV1.1) screener. Screen positives were evaluated on ASRSv1.1 symptom checklist, World Mental Health Survey Initiative Version of the World Health Organization Composite International Diagnostic Interview (WMH-CIDI) and by clinical assessments. DSM-IV-TR and Wender-Utah criteria were used for diagnosis. RESULTS: Twenty five men (mean age +/- SD, 23.4 +/- 5.6 yr) were diagnosed as having adult ADHD. The subjects' most common presentations were of losing temper and poor academic performance. The most common ADHD symptoms were difficulty in sustaining attention, easy distractibility, often losing things, and blurting out answers. A majority (80%) of the subjects had one or more psychiatric co-morbidity like oppositional defiant disorder, major depressive disorder and substance abuse/dependence. INTERPRETATION & CONCLUSION: Adult ADHD can be diagnosed in an Indian psychiatry outpatient setting. ADHD adults infrequently presented with the core symptoms of the disorder and had high psychiatric co-morbidity rates. PMID- 19287064 TI - Expression of class III beta-tubulin in colorectal carcinomas: an immunohistochemical study using TU-20 & TuJ-1 antibody. AB - BACKGROUND & OBJECTIVE: Expression of class III beta-tubulin represents newly discovered marker of resistance to taxol-based chemotherapy in a wide spectra of carcinomas. However, very little is known about its expression in colorectal carcinomas. This study was done to determine class III beta-tubulin expression in a large series of colonic carcinomas, covering tumours with different degree of differentiation in order to evaluate its prospective significance in resistance to taxol-based chemotherapeutics and to compare the immunostaining profile of two widely used monoclonal antibodies, TU-20 and TuJ-1 METHODS: Sixty patients with colorectal carcinoma were enrolled; all of them were treated surgically by the resection. Twenty tumours were histologically assessed as G1, 20 as G2 and 20 as G3. Routine immunohistochemical procedure using TU-20 and TuJ-1 mouse monoclonal antibodies was applied to all 60 specimen and slides were evaluated using an optical microscope. RESULTS: Expression of class III beta-tubulin was detected in 14 tumours (23.3%), while remaining tumours were negative. Relatively higher frequency of class III beta-tubulin expression was observed in G3 tumours (10 cases) in comparison with G1 (3 cases) and G2 (1 case), respectively. Seven tumours displayed positive immunostaining with both tested antibodies TU-20 and TuJ-1. Six tumours showed expression of class III beta- tubulin in more than 1 per cent of neoplastic cell population. In remaining 8 tumours only individual scattered neoplastic cells exhibited class III beta-tubulin expression either with TU-20, or with TuJ-1 antibody. INTERPRETATION & CONCLUSION: Higher frequency of immunoreactivity was observed in poorly differentiated tumours. However, more than 90 per cent of neoplastic cell population did not express class III beta tubulin in almost all tumours. These negative cells of colonic cancer could represent the potential target for taxane-based chemotherapy in the future. Our results indicate that TU-20 and TuJ-1 antibodies exhibit very similar immunoreactivity in neoplastic tissue. PMID- 19287065 TI - An increase in enteric fever cases due to Salmonella Paratyphi A in & around Chandigarh. AB - BACKGROUND & OBJECTIVE: Enteric fever is a major public health problem in India. It is classically caused by Salmonella enterica serotype Typhi. Salmonella enterica serotype Paratyphi A which had been reported less frequently from cases of enteric fever has shown an increasing trend since 1996 in India. There is also variation in the antimicrobial susceptibility of Salmonella Paratyphi A from different parts of the country. An attempt is therefore made to study the rate of isolation and antimicrobial susceptibility pattern of Salmonella Paratyphi A from cases of enteric fever coming to a tertiary care hospital at Chandigarh. METHODS: The blood samples of patients suspected of having enteric fever and admitted to Government Medical College and Hospital, Chandigarh, from January 2006 to April 2007 (11,240) were processed by conventional methods. Antimicrobial susceptibility was tested by Kirby-Bauer disc diffusion method. The minimum inhibitory concentration to two antibiotics- ciprofloxacin and chloramphenicol was determined by agar dilution technique. Simultaneously, retrospective analysis was done from January 2003-December 2005 to study any difference in the incidence and antimicrobial susceptibility pattern of Salmonella Paratyphi A among enteric fever patients. RESULTS: Of 305 total isolates, 231 were S. Typhi and 84 S. Paratyphi A rise. The number of Salmonella Paratyphi A cases rose from 27 in 2006 (34.18%) to 13 (40.63%) in four months of 2007. All were sensitive to ciprofloxacin and cefotaxime but MIC to ciprofloxacin was raised (0.125-0.5 microg/ml). Resistance to nalidixic acid was 92.5 per cent. Chloramphenicol sensitivity re-emerged with 90 per cent isolates sensitive to it while sensitivity to ampicillin dropped (72.5%) as compared to previous years. Only one isolate was multi-drug resistant. INTERPRETATION & CONCLUSION: The present study conferencing Salmonella Paratyphi A as the rapidly emerging pathogen of enteric fever. With increasing resistance to fluoroquinolones and possibility of re emergence of sensitivity to chloramphenicol, the policy of empirical treatment of enteric fever needs to be rationalized. PMID- 19287066 TI - Addition of isovitalex in chocolate agar for the isolation of Haemophilus influenzae. AB - BACKGROUND & OBJECTIVE: The reason for lack of data on burden of Haemophilus influenzae type b (Hib) in developing countries was mainly failure of detection of this fastidious organism in laboratories. Use of isovitalex (IVX) was suggested as an essential supplement for growing this organism. This study was carried out to investigate the impact of IVX supplementation to chocolate agar for detection of Hib. METHODS: Chocolate agar with and without supplementation of IVX was prepared. Clinical samples as well as reference strains of Hib were simultaneously cultured on both the media. RESULTS: H. influenzae isolates (N=194) were simultaneously grown on chocolate agar (CA) with and without isovitalex (IVX). Average colony size of H. influenzae on CA with IVX (CA-IVX) was larger only by 0.10 cm (range 0.05 to 0.16 cm) compared to CA alone. Addition of IVX to CA increased the cost of media by 2.1-fold. INTERPRETATION & CONCLUSION: Isovitalex is not essential for the isolation and growth of H. influenzae almost halving the cost. PMID- 19287067 TI - Newer strategies for the kala-azar elimination programme in India. PMID- 19287068 TI - PCR on formalin-fixed necropsy tissues to diagnose leptospirosis. PMID- 19287069 TI - Association of mutations in rpsL gene with high degree of streptomycin resistance in clinical isolates of Mycobacterium tuberculosis in India. PMID- 19287070 TI - The protein expression of TRP-1 and galectin-1 in cutaneous malignant melanomas. AB - BACKGROUND: Patients with metastazing malignant melanoma have a poor outcome and determination of thickness of the primary tumor remains as the most important prognostic predictor. The aim of this study was to use an antibody-based proteomics strategy to search for new molecular markers associated with melanoma progression. Two proteins, TRP-1 and galectin-1, were identified as proteins with enhanced expression in cells from the melanocytic lineage. PATIENTS AND METHODS: Protein profiling of TRP-1 and galectin-1 together with proliferation marker Ki 67 and melanocyte marker Melan-A was performed in normal tissues from 144 individuals and in 216 different tumors using tissue microarrays and immunohistochemistry. The protein expression pattern was further analyzed in a defined cohort of 157 patients diagnosed with invasive cutaneous malignant melanoma. RESULTS: Both TRP-1 and galectin-1 were highly expressed in normal melanocytes and melanoma. The expression of TRP-1 was inversely correlated with tumor stage (p=0.002, (R=-0.28)). Neither TRP-1 or galectin-1 was associated with overall or disease free survival (p>0.14, p>0.46 respectively). Ki-67 was associated with tumor stage and survival (p<0.001). CONCLUSION: TRP-1 and galectin-1 protein expression patterns were determined in normal and cancer tissues and both proteins were expressed in the majority of the malignant melanomas. There was no correlation between TRP-1 or galectin-1 expression and survival. PMID- 19287071 TI - Genomic analysis of prostate cancer stem cells isolated from a highly metastatic cell line. AB - BACKGROUND: Tumor-initiating or cancer stem cells (CSCs) were recently isolated from all major human cancers, including prostate cancer. However, the extreme heterogeneity of tumor cells in terms of biological behavior and gene expression patterns and difficulties isolating a pure population of CSCs from tumor tissues significantly impede a comparative analysis of CSCs. MATERIALS AND METHODS: Different phenotypic populations were isolated from a metastatic derivative of PC 3 cell line, PC3-MM2, and tested for their ability to form tumors in NOD/SCID mice and floating spheroids in 3D culture systems. RESULTS: All tested cell lines possessed minor populations of cells with highest expression of CD133, CD44 and CD166, whereas the vast majority of cells were CD133-negative. Several experimental approaches promoted a higher proportion of CD133-positive cells with increased in vivo tumorigenicity and the ability to produce floating spheres. Genome-wide microarray analysis (Affymetrix; DAVID) of CSC-enriched versus CSC depleted cell populations revealed 213 genes with 10-100 fold increased activity out of 8994 differentially expressed ones and 87 genes with 5-50 fold decreased activity. CONCLUSIONS: The proposed in vitro prostate CSC model allows for reliable isolation and propagation of highly tumorigenic cells. This study may contribute to the identification of novel targets for CSC-targeted prostate cancer treatment. PMID- 19287072 TI - Adenovirus-mediated thymidine kinase gene therapy and coxsackie adenovirus receptor expression in ovarian cancer cells. AB - Coxsackie adenovirus receptor (CAR) expression is the main mechanism of adenovirus entry into target cells. It is unclear whether CAR expression itself is influenced by transduction with the adenovirus-Rous sarcoma virus-thymidine kinase (ADV-RSV-TK) gene therapy construct or by the subsequent intracellular accumulation of the TK gene product. Antibody generation and characterization, immunocytochemistry, Western blotting and 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide (MTT) assay were performed to investigate the relationship of gene transfer and CAR expression as well as differences in therapeutic susceptibility of MDAH-2774 and OVCAR-3 cell lines to ADV-RSV-TK gene therapy. CAR expression was observed on the membranes but intracellular translocation of CAR also took place dependent on cellular growth patterns. TK gene expression was dependent on multiplicity of infection (MOI) and thus on vector dose in a linear fashion. Neither TK expression nor ADV transduction influenced CAR expression, or ADV-RSV-TK transduction. Differential susceptibility of different cell lines to TK-induced cell killing by acyclovir metabolites was observed. CAR expression appears not to be influenced by adenoviral transduction or by the accumulation of the TK gene product. Differences in therapeutic sensitivity are most likely mediated by intracellular mechanisms and not by modulation of CAR expression. PMID- 19287073 TI - Ochratoxin a lowers mRNA levels of genes encoding for key proteins of liver cell metabolism. AB - Ochratoxin A (OTA) is a nephro- and hepatotoxic mycotoxin that frequently contaminates food and feedstuffs. Although recent studies have indicated that OTA modulates renal gene expression, little is known regarding its impact on differential gene expression in the liver. Therefore a microarray study of the HepG2 liver cell transcriptome in response to OTA exposure (0, 0.25, 2.5 micromol/l for 24 h) was performed using Affymetrix GeneChip technology. Selected microarray results were verified by real-time PCR and Western blotting as independent methods. Out of 14,500 genes present on the microarray, 13 and 250 genes were down-regulated by 0.25 and 2.5 micromol/l OTA, respectively. Reduced mRNA levels of calcineurin A beta (PPP3CB), which regulates inflammatory signalling pathways in immune cells, and of the uncoupling protein 2 (UCP2), which has been suggested to control the production of reactive oxygen species (ROS), were observed in response to 0.25 micromol/l OTA. A particularly strong down-regulation due to 2.5 micromol/l OTA was evident for the mRNA levels of insulin-like growth factor binding protein 1 (IGFBP1) and tubulin beta 1 (TUBB1) which have been demonstrated to function as a pro-survival factor in hepatocytes and as an important cytoskeletal component, respectively. In addition, many genes involved in energy and xenobiotic metabolism, including phosphoglycerate kinase 1 (PGK1), stearoyl-Coenzyme A desaturase 1 (SCD), and glutathione S-transferase omega 1 (GSTO1), were down-regulated by OTA. Furthermore, OTA significantly inhibited the capacitative calcium entry into the HepG2 cells, indicating an alteration of calcium homeostasis. Overall, OTA dose-dependently affects multiple genes encoding for key proteins of liver cell metabolism. PMID- 19287074 TI - Prognostic utility of glycosyltransferase expression in breast cancer. AB - BACKGROUND: The post-translational modification of proteins, including glycosylation, is known to differ between normal and tumour cells. In this study, the expression profile of two glycosyltranferases, UDP-N-acetyl-D-galactosamine: polypeptide N-acetylgalactosaminyltransferase-6 (ppGalNAc-T6) and alpha6-sialyl transferase-I (ST6GalNAc-I) was assessed, in a cohort of women with breast cancer. PATIENTS AND METHODS: Breast cancer tissues (n=127) and normal background tissues (n=33) were collected immediately after excision during surgery. Following RNA extraction, reverse transcription was carried out and transcript levels were determined using real-time quantitative PCR and normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. Transcript levels within the breast cancer specimens were compared to the normal background tissues and analyzed against conventional pathological parameters and clinical outcome over a 10 year follow-up period. RESULTS: Significantly higher levels of ppGalNAc T6 were found in the breast cancer specimens compared to the background tissue (p=0.015). There was a non-significant trend for levels to increase with the Nottingham Prognostic Index (NPI) and TNM stage and those who died from breast cancer. ST6GalNAc-I expression was associated with better prognosis, reaching significance when comparing patients who remained disease free to those with distant recurrence (p=0.0096). The relationship approached significance when comparing NPI 2 to NPI 3 (p=0.058) and disease free patients to non-disease free patients (p=0.052) or those who died of breast cancer (p=0.060). For both enzymes a significant association with ductal type was found. CONCLUSION: Expression of ppGalNAc-T6 is significantly higher in breast cancer compared to 'normal'/benign breast tissue samples. ST6GalNAc-I expression in breast cancer is associated with better prognosis. PMID- 19287075 TI - Is there an influence of the surrounding material on the response of the alanine dosimetry system? AB - In a combined experimental and Monte Carlo study the possible influence of the surrounding material on the response of the alanine dosimetry system was investigated. The aim of this work was to estimate the uncertainties induced by the surroundings with respect to quality assurance measurements for radiotherapy, for example in humanoid phantoms. Six different materials were tested. The electron density range covered comprises the range present in human tissue. No significant influence of the surrounding material could be found for irradiations in the (60)Co reference field of the Physikalisch-Technische Bundesanstalt (PTB). PMID- 19287076 TI - Design and evaluation of a probe for simultaneous EEG and near-infrared imaging of cortical activation. AB - We present a novel probe design which enables simultaneous electroencephalography (EEG) and near-infrared (NIR) optical imaging to be performed in a manner which is easy to apply, allows for optimum co-registration of the two forms of data and maximizes the number of sensors which can be applied to a given area. Our probe design is evaluated using a dual-modality, tissue-mimicking phantom and by performing a simple functional activation study of the human motor cortex. We successfully acquired NIR optical and EEG data simultaneously for both our phantom and our human motor cortex experiments, clearly demonstrating the effectiveness and suitability of our 'opto-electrode'. PMID- 19287077 TI - Functional localization and visualization of the subthalamic nucleus from microelectrode recordings acquired during DBS surgery with unsupervised machine learning. AB - Microelectrode recordings are a useful adjunctive method for subthalamic nucleus localization during deep brain stimulation surgery for Parkinson's disease. Attempts to quantitate and standardize this process, using single computational measures of neural activity, have been limited by variability in patient neurophysiology and recording conditions. Investigators have suggested that a multi-feature approach may be necessary for automated approaches to perform within acceptable clinical standards. We present a novel data visualization algorithm and several unique features that address these shortcomings. The algorithm extracts multiple computational features from the microelectrode neurophysiology and integrates them with tools from unsupervised machine learning. The resulting colour-coded map of neural activity reveals activity transitions that correspond to the anatomic boundaries of subcortical structures. Using these maps, a non-neurophysiologist is able to achieve sensitivities of 90% and 95% for STN entry and exit, respectively, to within 0.5 mm accuracy of the current gold standard. The accuracy of this technique is attributed to the multi feature approach. This activity map can simplify and standardize the process of localizing the subthalamic nucleus (STN) for neurostimulation. Because this method does not require a stationary electrode for careful recording of unit activity for spike sorting, the length of the operation may be shortened. PMID- 19287078 TI - In vivo evaluation of a neural stem cell-seeded prosthesis. AB - Neural prosthetics capable of recording or stimulating neuronal activity may restore function for patients with motor and sensory deficits resulting from injury or degenerative disease. However, overcoming inconsistent recording quality and stability in chronic applications remains a significant challenge. A likely reason for this is the reactive tissue response to the devices following implantation into the brain, which is characterized by neuronal loss and glial encapsulation. We have developed a neural stem cell-seeded probe to facilitate integration of a synthetic prosthesis with the surrounding brain tissue. We fabricated parylene devices that include an open well seeded with neural stem cells encapsulated in an alginate hydrogel scaffold. Quantitative and qualitative data describing the distribution of neuronal, glial, and progenitor cells surrounding seeded and control devices are reported over four time points spanning 3 months. Neuronal loss and glial encapsulation associated with cell seeded probes were mitigated during the initial week of implantation and exacerbated by 6 weeks post-insertion compared to control conditions. We hypothesize that graft cells secrete neuroprotective and neurotrophic factors that effect the desired healing response early in the study, with subsequent cell death and scaffold degradation accounting for a reversal of these results later. Applications of this biohybrid technology include future long-term neural recording and sensing studies. PMID- 19287079 TI - A fully automated non-external marker 4D-CT sorting algorithm using a serial cine scanning protocol. AB - Current 4D-CT methods require external marker data to retrospectively sort image data and generate CT volumes. In this work we develop an automated 4D-CT sorting algorithm that performs without the aid of data collected from an external respiratory surrogate. The sorting algorithm requires an overlapping cine scan protocol. The overlapping protocol provides a spatial link between couch positions. Beginning with a starting scan position, images from the adjacent scan position (which spatial match the starting scan position) are selected by maximizing the normalized cross correlation (NCC) of the images at the overlapping slice position. The process was continued by 'daisy chaining' all couch positions using the selected images until an entire 3D volume was produced. The algorithm produced 16 phase volumes to complete a 4D-CT dataset. Additional 4D-CT datasets were also produced using external marker amplitude and phase angle sorting methods. The image quality of the volumes produced by the different methods was quantified by calculating the mean difference of the sorted overlapping slices from adjacent couch positions. The NCC sorted images showed a significant decrease in the mean difference (p < 0.01) for the five patients. PMID- 19287080 TI - The relevance of very low energy ions for heavy-ion therapy. AB - Heavy-ion radiotherapy exploits the high biological effectiveness of localized energy deposition delivered by so-called Bragg-peak particles. Recent publications have challenged the established procedures to calculate biological effective dose distributions in treatment planning. They emphasize the importance of very low energy (<500 keV amu(-1)) ions, either as primary particles or originating from molecular and nuclear fragmentations. We show, however, that slow heavy ions with energies below 500 keV amu(-1) only play a negligible role in cancer treatments for several reasons. Their residual range is very small compared to the relevant length scale of treatment planning. Moreover, their relative frequency and also their relative dose distribution are insignificant, since energy loss and range straggling in ion slowing down processes as well as the necessary superposition of Bragg peaks wash out small-scale special effects. Additionally, we show that even a 1000 times larger biological damage of such slow ions would not result in a clinically relevant increase of the photon equivalent dose. Therefore, neither a more precise physical description of ions in the very distal part of the Bragg peak nor the consideration of radiation damage induced by hyperthermal ions would result in a meaningful improvement of current models for heavy-ion treatment planning. PMID- 19287081 TI - Real-time MRI-guided hyperthermia treatment using a fast adaptive algorithm. AB - Magnetic resonance (MR) imaging is promising for monitoring and guiding hyperthermia treatments. The goal of this work is to investigate the stability of an algorithm for online MR thermal image guided steering and focusing of heat into the target volume. The control platform comprised a four-antenna mini annular phased array (MAPA) applicator operating at 140 MHz (used for extremity sarcoma heating) and a GE Signa Excite 1.5 T MR system, both of which were driven by a control workstation. MR proton resonance frequency shift images acquired during heating were used to iteratively update a model of the heated object, starting with an initial finite element computed model estimate. At each iterative step, the current model was used to compute a focusing vector, which was then used to drive the next iteration, until convergence. Perturbation of the driving vector was used to prevent the process from stalling away from the desired focus. Experimental validation of the performance of the automatic treatment platform was conducted with two cylindrical phantom studies, one homogeneous and one muscle equivalent with tumor tissue (conductivity 50% higher) inserted, with initial focal spots being intentionally rotated 90 degrees and 50 degrees away from the desired focus, mimicking initial setup errors in applicator rotation. The integrated MR-HT treatment platform steered the focus of heating into the desired target volume in two quite different phantom tissue loads which model expected patient treatment configurations. For the homogeneous phantom test where the target was intentionally offset by 90 degrees rotation of the applicator, convergence to the proper phase focus in the target occurred after 16 iterations of the algorithm. For the more realistic test with a muscle equivalent phantom with tumor inserted with 50 degrees applicator displacement, only two iterations were necessary to steer the focus into the tumor target. Convergence improved the heating efficacy (the ratio of integral temperature in the tumor to integral temperature in normal tissue) by up to six-fold, compared to the first iteration. The integrated MR-HT treatment algorithm successfully steered the focus of heating into the desired target volume for both the simple homogeneous and the more challenging muscle equivalent phantom with tumor insert models of human extremity sarcomas after 16 and 2 iterations, correspondingly. The adaptive method for MR thermal image guided focal steering shows promise when tested in phantom experiments on a four-antenna phased array applicator. PMID- 19287082 TI - Simulating oblique incident irradiation using the BEAMnrc Monte Carlo code. AB - A new source for the simulation of oblique incident irradiation has been developed for the BEAMnrc Monte Carlo code. In this work, we describe a method for the simulation of any component that is rotated at some angle relative to the central axis of the modelled radiation unit. The performance of the new BEAMnrc source was validated against experimental measurements. The comparison with ion chamber data showed very good agreement between experiments and calculation for a number of oblique irradiation angles ranging from 0 degrees to 30 degrees . The routine was also cross-validated, in geometrically equivalent conditions, against a different radiation source available in the DOSXYZnrc code. The test showed excellent consistency between the two routines. The new radiation source can be particularly useful for the Monte Carlo simulation of radiation units in which the radiation beam is tilted with respect to the unit's central axis. To highlight this, a modern cone-beam CT unit is modelled using this new source and validated against measurement. PMID- 19287083 TI - Quantifying lung morphology with respiratory-gated micro-CT in a murine model of emphysema. AB - Non-invasive micro-CT imaging techniques have been developed to investigate lung structure in free-breathing rodents. In this study, we investigate the utility of retrospectively respiratory-gated micro-CT imaging in an emphysema model to determine if anatomical changes could be observed in the image-derived quantitative analysis at two respiratory phases. The emphysema model chosen was a well-characterized, genetically altered model (TIMP-3 knockout mice) that exhibits a homogeneous phenotype. Micro-CT scans of the free-breathing, anaesthetized mice were obtained in 50 s and retrospectively respiratory sorted and reconstructed, providing 3D images representing peak inspiration and end expiration with 0.15 mm isotropic voxel spacing. Anatomical measurements included the volume and CT density of the lungs and the volume of the major airways, along with the diameters of the trachea, left bronchus and right bronchus. From these measurements, functional parameters such as functional residual capacity and tidal volume were calculated. Significant differences between the wild-type and TIMP-3 knockout groups were observed for measurements of CT density over the entire lung, indicating increased air content in the lungs of TIMP-3 knockout mice. These results demonstrate retrospective respiratory-gated micro-CT, providing images at multiple respiratory phases that can be analyzed quantitatively to investigate anatomical changes in murine models of emphysema. PMID- 19287084 TI - Efficient gamma index calculation using fast Euclidean distance transform. AB - The gamma index is a tool for dose distribution comparison. It combines both dose difference (DD) and distance to agreement (DTA) into a single quantity. Though it is an effective measure, making up for the inadequacy of DD or DTA alone, its calculation can be very time-consuming. For a k-D space with N quantization levels in each dimension, the complexity of the exhaustive search is O(N(2k)). In this work, we proposed an efficient method that reduces the complexity from O(N(2k)) to O(N(k)M), where M is the number of discretized dose values and is comparable to N. More precisely, by embedding the reference dose distribution in a (k+1)-D spatial-dose space, we can use fast Euclidean distance transform with linear complexity to obtain a table of gamma indices evaluated over a range of the (k+1)-D spatial-dose space. Then, to obtain gamma indices for the test dose distribution, it requires only table lookup with complexity O(N(k)). Such a table can also be used for other test dose distributions as long as the reference dose distribution is the same. Simulations demonstrated the efficiency of our proposed method. The speedup for 3D gamma index calculation is expected to be on the order of tens of thousands (from O(N(6)) to O(N(3)M)) if N is a few hundreds, which makes clinical usage of the 3D gamma index feasible. A byproduct of the gamma index table is that the gradient of the gamma index with respect to either the spatial or dose dimension can be easily derived. The gradient can be used to identify the main causes of the discrepancy from the reference distribution at any dose point in the test distribution or incorporated in treatment planning and machine parameter optimization. PMID- 19287085 TI - Dynamic splitting of Gaussian pencil beams in heterogeneity-correction algorithms for radiotherapy with heavy charged particles. AB - The pencil-beam algorithm is valid only when elementary Gaussian beams are small enough compared to the lateral heterogeneity of a medium, which is not always true in actual radiotherapy with protons and ions. This work addresses a solution for the problem. We found approximate self-similarity of Gaussian distributions, with which Gaussian beams can split into narrower and deflecting daughter beams when their sizes have overreached lateral heterogeneity in the beam-transport calculation. The effectiveness was assessed in a carbon-ion beam experiment in the presence of steep range compensation, where the splitting calculation reproduced a detour effect amounting to about 10% in dose or as large as the lateral particle disequilibrium effect. The efficiency was analyzed in calculations for carbon-ion and proton radiations with a heterogeneous phantom model, where the beam splitting increased computing times by factors of 4.7 and 3.2. The present method generally improves the accuracy of the pencil-beam algorithm without severe inefficiency. It will therefore be useful for treatment planning and potentially other demanding applications. PMID- 19287086 TI - In vitro kinetic studies on the mechanism of oxygen-dependent cellular uptake of copper radiopharmaceuticals. AB - The development of hypoxia-selective radiopharmaceuticals for use as therapeutic and/or imaging agents is of vital importance for both early identification and treatment of cancer and in the design of new drugs. Radiotracers based on copper for use in positron emission tomography have received great attention due to the successful application of copper(II) bis(thiosemicarbazonato) complexes, such as [(60/62/64)Cu(II)ATSM] and [(60/62/64)Cu(II)PTSM], as markers for tumour hypoxia and blood perfusion, respectively. Recent work has led to the proposal of a revised mechanism of hypoxia-selective cellular uptake and retention of [Cu(II)ATSM]. The work presented here describes non-steady-state kinetic simulations in which the reported pO(2)-dependent in vitro cellular uptake and retention of [(64)Cu(II)ATSM] in EMT6 murine carcinoma cells has been modelled by using the revised mechanistic scheme. Non-steady-state (NSS) kinetic analysis reveals that the model is in very good agreement with the reported experimental data with a root-mean-squared error of less than 6% between the simulated and experimental cellular uptake profiles. Estimated rate constants are derived for the cellular uptake and washout (k(1) = 9.8 +/- 0.59 x 10(-4) s(-1) and k(2) = 2.9 +/- 0.17 x 10(-3) s(-1)), intracellular reduction (k(3) = 5.2 +/- 0.31 x 10( 2) s(-1)), reoxidation (k(4) = 2.2 +/- 0.13 mol(-1) dm(3) s(-1)) and proton mediated ligand dissociation (k(5) = 9.0 +/- 0.54 x 10(-5) s(-1)). Previous mechanisms focused on the reduction and reoxidation steps. However, the data suggest that the origins of hypoxia-selective retention may reside with the stability of the copper(I) anion with respect to protonation and ligand dissociation. In vitro kinetic studies using the nicotimamide adenine dinucleotide (NADH)-dependent ferredoxin reductase enzyme PuR isolated from the bacterium Rhodopseudomonas palustris have also been conducted. NADH turnover frequencies are found to be dependent on the structure of the ligand and the results confirm that the proposed reduction step in the mechanism of hypoxia selectivity is likely to be mediated by NADH-dependent enzymes. Further understanding of the mechanism of hypoxia selectivity may facilitate the development of new imaging and radiotherapeutic agents with increased specificity for tumour hypoxia. PMID- 19287087 TI - The adaptation of megavoltage cone beam CT for use in standard radiotherapy treatment planning. AB - Potential areas where megavoltage computed tomography (MVCT) could be used are second- and third-phase treatment planning in 3D conformal radiotherapy and IMRT, adaptive radiation therapy, single fraction palliative treatment and for the treatment of patients with metal prostheses. A feasibility study was done on using MV cone beam CT (CBCT) images generated by proprietary 3D reconstruction software based on the FDK algorithm for megavoltage treatment planning. The reconstructed images were converted to a DICOM file set. The pixel values of megavoltage cone beam computed tomography (MV CBCT) were rescaled to those of kV CT for use with a treatment planning system. A calibration phantom was designed and developed for verification of geometric accuracy and CT number calibration. The distance measured between two marker points on the CBCT image and the physical dimension on the phantom were in good agreement. Point dose verification for a 10 cm x 10 cm beam at a gantry angle of 0 degrees and SAD of 100 cm were performed for a 6 MV beam for both kV and MV CBCT images. The point doses were found to vary between +/-6.1% of the dose calculated from the kV CT image. The isodose curves for 6 MV for both kV CT and MV CBCT images were within 2% and 3 mm distance-to-agreement. A plan with three beams was performed on MV CBCT, simulating a treatment plan for cancer of the pituitary. The distribution obtained was compared with those corresponding to that obtained using the kV CT. This study has shown that treatment planning with MV cone beam CT images is feasible. PMID- 19287088 TI - In vivo micro-CT analysis of bone remodeling in a rat calvarial defect model. AB - The rodent calvarial defect model is commonly used to investigate bone regeneration and wound healing. This study presents a micro-computed tomography (micro-CT) methodology for measuring the bone mineral content (BMC) in a rat calvarial defect and validates it by estimating its precision error. Two defect models were implemented. A single 6 mm diameter defect was created in 20 rats, which were imaged in vivo for longitudinal experiments. Three 5 mm diameter defects were created in three additional rats, which were repeatedly imaged ex vivo to determine precision. Four control rats and four rats treated with bone morphogenetic protein were imaged at 3, 6, 9 and 12 weeks post-surgery. Scan parameters were 80 kVp, 0.45 mA and 180 mAs. Images were reconstructed with an isotropic resolution of 45 microm. At 6 weeks, the BMC in control animals (4.37 +/- 0.66 mg) was significantly lower (p < 0.05) than that in treated rats (11.29 +/- 1.01 mg). Linear regression between the BMC and bone fractional area, from 20 rats, showed a strong correlation (r(2) = 0.70, p < 0.0001), indicating that the BMC can be used, in place of previous destructive analysis techniques, to characterize bone growth. The high precision (2.5%) of the micro-CT methodology indicates its utility in detecting small BMC changes in animals. PMID- 19287089 TI - A feasibility study of novel plastic scintillation dosimetry with pulse-counting mode. AB - The purpose of this study was to develop a novel scintillation dosimeter for in vivo dosimetry in Ir-192 brachytherapy via the pulse-counting mode. The new dosimeter was made from a plastic scintillator shaped into a hemisphere of diameter 1 mm and connected to the tip of a plastic optical fiber. The relationship between pulse counts and absorbed dose was derived based on the assumption that scintillation photons from the incident gamma ray are proportional to the absorbed dose. An equation for the conversion of pulse counts to water-equivalent dose was deduced wherein the pulse height spectrum from scintillation photons was assumed to be exponential. To confirm its accuracy, the dose rate distribution in a water phantom was measured by the present dosimeter and this was compared with Monte Carlo simulations, resulting in a discrepancy of less than 1.97%. It was found that the dosimeter has a wide dynamic range of linearity up to an order of magnitude of almost 10(3), including corrections for loss of counts due to pile-up. PMID- 19287090 TI - Enhanced T cell recovery in HIV-1-infected adults through IL-7 treatment. AB - HIV infection results in CD4+ T cell deficiency, but efficient combination antiretroviral therapy (c-ART) restores T cells and decreases morbidity and mortality. However, immune restoration by c-ART remains variable, and prolonged T cell deficiency remains in a substantial proportion of patients. In a prospective open-label phase I/IIa trial, we evaluated the safety and efficacy of administration of the T cell regulator IL-7. The trial included 13 c-ART-treated HIV-infected patients whose CD4+ cell counts were between 100 and 400 cells/microl and plasma HIV RNA levels were less than 50 copies/ml. Patients received a total of 8 subcutaneous injections of 2 different doses of recombinant human IL-7 (rhIL-7; 3 or 10 microg/kg) 3 times per week over a 16-day period. rhIL-7 was well tolerated and induced a sustained increase of naive and central memory CD4+ and CD8+ T cells. In the highest dose group, 4 patients experienced transient increases in viral replication. However, functional assays showed that the expanded T cells responded to HIV antigen by producing IFN-gamma and/or IL-2. In conclusion, in lymphopenic HIV-infected patients, rhIL-7 therapy induced substantial functional and quantitative changes in T cells for 48 weeks. Therefore, patients may benefit from intermittent therapy with IL-7 in combination with c-ART. PMID- 19287091 TI - Macrophage deficiency of p38alpha MAPK promotes apoptosis and plaque necrosis in advanced atherosclerotic lesions in mice. AB - ER stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. Therefore, signaling pathways that alter ER stress-induced apoptosis may affect advanced atherosclerosis. Here we placed Apoe-/- mice deficient in macrophage p38alpha MAPK on a Western diet and found that they had a marked increase in macrophage apoptosis and plaque necrosis. The macrophage p38alpha-deficient lesions also exhibited a significant reduction in collagen content and a marked thinning of the fibrous cap, which suggests that plaque progression was advanced in these mice. Consistent with our in vivo data, we found that ER stress-induced apoptosis in cultured primary mouse macrophages was markedly accelerated under conditions of p38 inhibition. Pharmacological inhibition or genetic ablation of p38 suppressed activation of Akt in cultured macrophages and in atherosclerotic lesions. In addition, inhibition of Akt enhanced ER stress-induced macrophage apoptosis, and expression of a constitutively active myristoylated Akt blocked the enhancement of ER stress-induced apoptosis that occurred with p38 inhibition in cultured cells. Our results demonstrate that p38alpha MAPK may play a critical role in suppressing ER stress-induced macrophage apoptosis in vitro and advanced lesional macrophage apoptosis in vivo. PMID- 19287092 TI - Loss-of-function variants in endothelial lipase are a cause of elevated HDL cholesterol in humans. AB - Elevated plasma concentrations of HDL cholesterol (HDL-C) are associated with protection from atherosclerotic cardiovascular disease. Animal models indicate that decreased expression of endothelial lipase (LIPG) is inversely associated with HDL-C levels, and genome-wide association studies have identified LIPG variants as being associated with HDL-C levels in humans. We hypothesized that loss-of-function mutations in LIPG may result in elevated HDL-C and therefore performed deep resequencing of LIPG exons in cases with elevated HDL-C levels and controls with decreased HDL-C levels. We identified a significant excess of nonsynonymous LIPG variants unique to cases with elevated HDL-C. In vitro lipase activity assays demonstrated that these variants significantly decreased endothelial lipase activity. In addition, a meta-analysis across 5 cohorts demonstrated that the low-frequency Asn396Ser variant is significantly associated with increased HDL-C, while the common Thr111Ile variant is not. Functional analysis confirmed that the Asn396Ser variant has significantly decreased lipase activity both in vitro and in vivo, while the Thr111Ile variant has normal lipase activity. Our results establish that loss-of-function mutations in LIPG lead to increased HDL-C levels and support the idea that inhibition of endothelial lipase may be an effective mechanism to raise HDL-C. PMID- 19287093 TI - Plasma membrane Ca2+-ATPase isoform 4 antagonizes cardiac hypertrophy in association with calcineurin inhibition in rodents. AB - How Ca2+-dependent signaling effectors are regulated in cardiomyocytes, given the extreme cytoplasmic Ca2+ concentration changes that underlie contraction, remains unknown. Cardiomyocyte plasma membrane Ca2+-ATPase (PMCA) extrudes Ca2+ but has little effect on excitation-contraction coupling, suggesting its potential role in controlling Ca2+-dependent signaling effectors such as calcineurin. We generated cardiac-specific inducible PMCA4b transgenic mice that displayed normal global Ca2+ transient and cellular contraction levels and reduced cardiac hypertrophy following transverse aortic constriction (TAC) or phenylephrine/Ang II infusion, but showed no reduction in exercise-induced hypertrophy. Transgenic mice were protected from decompensation and fibrosis following long-term TAC. The PMCA4b transgene reduced the hypertrophic augmentation associated with transient receptor potential canonical 3 channel overexpression, but not that associated with activated calcineurin. Furthermore, Pmca4 gene-targeted mice showed increased cardiac hypertrophy and heart failure events after TAC. Physical associations between PMCA4b and calcineurin were enhanced by TAC and by agonist stimulation of cultured neonatal cardiomyocytes. PMCA4b reduced calcineurin nuclear factor of activated T cell-luciferase activity after TAC and in cultured neonatal cardiomyocytes after agonist stimulation. PMCA4b overexpression inhibited cultured cardiomyocyte hypertrophy following agonist stimulation, but much less so in a Ca2+ pumping-deficient PMCA4b mutant. Thus, Pmca4b likely reduces the local Ca2+ signals involved in reactive cardiomyocyte hypertrophy via calcineurin regulation. PMID- 19287094 TI - The TEL-AML1 leukemia fusion gene dysregulates the TGF-beta pathway in early B lineage progenitor cells. AB - Chromosome translocation to generate the TEL-AML1 (also known as ETV6-RUNX1) chimeric fusion gene is a frequent and early or initiating event in childhood acute lymphoblastic leukemia (ALL). Our starting hypothesis was that the TEL-AML1 protein generates and maintains preleukemic clones and that conversion to overt disease requires secondary genetic changes, possibly in the context of abnormal immune responses. Here, we show that a murine B cell progenitor cell line expressing inducible TEL-AML1 proliferates at a slower rate than parent cells but is more resistant to further inhibition of proliferation by TGF-beta. This facilitates the competitive expansion of TEL-AML1-expressing cells in the presence of TGF-beta. Further analysis indicated that TEL-AML1 binds to a principal TGF-beta signaling target, Smad3, and compromises its ability to activate target promoters. In mice expressing a TEL-AML1 transgene, early, pre pro-B cells were increased in number and also showed reduced sensitivity to TGF beta-mediated inhibition of proliferation. Moreover, expression of TEL-AML1 in human cord blood progenitor cells led to the expansion of a candidate preleukemic stem cell population that had an early B lineage phenotype (CD34+CD38-CD19+) and a marked growth advantage in the presence of TGF-beta. Collectively, these data suggest a plausible mechanism by which dysregulated immune responses to infection might promote the malignant evolution of TEL-AML1-expressing preleukemic clones. PMID- 19287095 TI - The OTT-MAL fusion oncogene activates RBPJ-mediated transcription and induces acute megakaryoblastic leukemia in a knockin mouse model. AB - Acute megakaryoblastic leukemia (AMKL) is a form of acute myeloid leukemia (AML) associated with a poor prognosis. The genetics and pathophysiology of AMKL are not well understood. We generated a knockin mouse model of the one twenty-two megakaryocytic acute leukemia (OTT-MAL) fusion oncogene that results from the t(1;22)(p13;q13) translocation specifically associated with a subtype of pediatric AMKL. We report here that OTT-MAL expression deregulated transcriptional activity of the canonical Notch signaling pathway transcription factor recombination signal binding protein for immunoglobulin kappa J region (RBPJ) and caused abnormal fetal megakaryopoiesis. Furthermore, cooperation between OTT-MAL and an activating mutation of the thrombopoietin receptor myeloproliferative leukemia virus oncogene (MPL) efficiently induced a short latency AMKL that recapitulated all the features of human AMKL, including megakaryoblast hyperproliferation and maturation block, thrombocytopenia, organomegaly, and extensive fibrosis. Our results establish that concomitant activation of RBPJ (Notch signaling) and MPL (cytokine signaling) transforms cells of the megakaryocytic lineage and suggest that specific targeting of these pathways could be of therapeutic value for human AMKL. PMID- 19287096 TI - Glomerular type 1 angiotensin receptors augment kidney injury and inflammation in murine autoimmune nephritis. AB - Studies in humans and animal models indicate a key contribution of angiotensin II to the pathogenesis of glomerular diseases. To examine the role of type 1 angiotensin (AT1) receptors in glomerular inflammation associated with autoimmune disease, we generated MRL-Faslpr/lpr (lpr) mice lacking the major murine type 1 angiotensin receptor (AT1A); lpr mice develop a generalized autoimmune disease with glomerulonephritis that resembles SLE. Surprisingly, AT1A deficiency was not protective against disease but instead substantially accelerated mortality, proteinuria, and kidney pathology. Increased disease severity was not a direct effect of immune cells, since transplantation of AT1A-deficient bone marrow did not affect survival. Moreover, autoimmune injury in extrarenal tissues, including skin, heart, and joints, was unaffected by AT1A deficiency. In murine systems, there is a second type 1 angiotensin receptor isoform, AT1B, and its expression is especially prominent in the renal glomerulus within podocytes. Further, expression of renin was enhanced in kidneys of AT1A-deficient lpr mice, and they showed evidence of exaggerated AT1B receptor activation, including substantially increased podocyte injury and expression of inflammatory mediators. Administration of losartan, which blocks all type 1 angiotensin receptors, reduced markers of kidney disease, including proteinuria, glomerular pathology, and cytokine mRNA expression. Since AT1A-deficient lpr mice had low blood pressure, these findings suggest that activation of type 1 angiotensin receptors in the glomerulus is sufficient to accelerate renal injury and inflammation in the absence of hypertension. PMID- 19287098 TI - Role of anticoagulation in the management of pancreatic cancer. AB - Pancreatic cancer remains a major clinical challenge. Recent advances in chemotherapeutic and targeted agents have offered a modest survival benefit. One of the major complications of pancreatic cancer is venous thromboembolism. Although it is well-known fact that patients with mucinous carcinoma of the pancreas and gastrointestinal tract pose an increased risk of developing thromboembolic complications, scarce data exists regarding the incidence and pathogenesis of venous thromboembolism in pancreatic cancer patients. The incidence of venous thromboembolism in pancreatic cancer patients ranges from 17% to 57%. Clinical data also suggest that the occurrence of venous thromboembolism may be associated with poorer prognosis in such patients. Recent data suggest that anticoagulant treatments may improve cancer patient survival by decreasing thromboembolic complications as well as by anticancer effects. Thromboembolic disease in pancreatic cancer presents a life-threatening complication and is regarded as paraneoplastic manifestation of the disease. Effective management of this risk factor is very important in the management of pancreatic cancer. Given the lack of extensive data and the clinical relevance of this topic for both physicians and basic research scientists, the authors review the incidence, pathogenesis and clinical implications of venous thromboembolism in pancreatic cancer patients. PMID- 19287097 TI - WNT1-inducible signaling protein-1 mediates pulmonary fibrosis in mice and is upregulated in humans with idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is characterized by distorted lung architecture and loss of respiratory function. Enhanced (myo)fibroblast activation, ECM deposition, and alveolar epithelial type II (ATII) cell dysfunction contribute to IPF pathogenesis. However, the molecular pathways linking ATII cell dysfunction with the development of fibrosis are poorly understood. Here, we demonstrate, in a mouse model of pulmonary fibrosis, increased proliferation and altered expression of components of the WNT/beta catenin signaling pathway in ATII cells. Further analysis revealed that expression of WNT1-inducible signaling protein-1 (WISP1), which is encoded by a WNT target gene, was increased in ATII cells in both a mouse model of pulmonary fibrosis and patients with IPF. Treatment of mouse primary ATII cells with recombinant WISP1 led to increased proliferation and epithelial-mesenchymal transition (EMT), while treatment of mouse and human lung fibroblasts with recombinant WISP1 enhanced deposition of ECM components. In the mouse model of pulmonary fibrosis, neutralizing mAbs specific for WISP1 reduced the expression of genes characteristic of fibrosis and reversed the expression of genes associated with EMT. More importantly, these changes in gene expression were associated with marked attenuation of lung fibrosis, including decreased collagen deposition and improved lung function and survival. Our study thus identifies WISP1 as a key regulator of ATII cell hyperplasia and plasticity as well as a potential therapeutic target for attenuation of pulmonary fibrosis. PMID- 19287099 TI - The Prevention, recognition and treatment of post-ERCP pancreatitis. PMID- 19287100 TI - Pancreatic cancer: translating lessons from mouse models. AB - Pancreatic cancer is the 10th most common cancer and 4th cause of cancer related deaths. Progress in diagnosis and treatment has been slow and disappointing but improvement in understanding of pathogenesis and of molecular changes may offer some ground for rational and etiological approach. During the last ten years the first evidence about the benefit of targeting dysregulated pathways was provided by the study that tried the addition of the EGFR inhibitor erlotinib to the standard cytotoxic gemcitabine. Since then, despite other numerous negative studies, various agents have been investigated in the preclinical and clinical setting and are currently through drug development pipeline. In this year's Gastrointestinal Symposium of the American Society of Clinical Oncology (GI ASCO, San Francisco, 15-17 January 2009), translational and clinical researchers presented evidence of specific genetic variations predicting toxicity (Abstract #115) or efficacy (Abstract #118) of gemcitabine-based treatment and of clinical biomarkers which may serve as predictors of therapy (Abstract #117) or mortality (Abstract #202). We were also informed about the presence of a new surface antigen (CD133) in pancreatic cancer stem cells (Abstract #150) and the development of a recombinant viral vector carrying the G antigen 1 (GAGE1) gene able for B-cells transduction (Abstract #178), which may lead potentially to the development of new immunotherapies and targeted agents. Study and efficacy of novel targeted molecules in preclinical models in vitro and in vivo was also presented (Abstracts #144, 145, 158, 163). In contrast to other malignancies, no mutations of the EGFR/PI3K pathway were found in pancreatic cancer cells not allowing thus a patients' selection approach for EGFR antibodies (Abstract #173). PMID- 19287101 TI - Screening for early pancreatic ductal adenocarcinoma: an urgent call! AB - Cancer of the pancreas is the fourth leading cause of cancer mortality in the United States. The annual death rate from the disease almost equals the annual incidence due to the aggressive nature of the cancer as well as to the lack of effective means of screening for it during its early curable stage. Molecular markers and imaging have not proven to be accurate modalities for screening for pancreatic cancer. The diagnosis and management of pancreatic cancer continues to be an overwhelming challenge. The authors discuss the current status of screening for pancreatic cancer and summarize relevant studies presented in the 2009 GI Cancers Symposium: utility of endoscopic ultrasound in screening high risk patients (Abstract #112), diagnostic performance of a highly specific antibody for MUC1 (Abstract #113), use of metabonomics for the early detection of pancreatic adenocarcinoma (Abstract #126), and a report on the potential impact of delay in diagnosis and treatment on pancreatic cancer outcomes at a tertiary care center (Abstract #137). PMID- 19287102 TI - Advancements in the management of pancreatic cancer. AB - Management of pancreatic cancer remains the most challenging work in oncology. Though pancreatic cancer represents only 2-3% of all cancers, it is the most fatal one accounting for the 6% of all cancer death. It remains the 4th cause of death by cancer since 1970s in the U.S.. Gemcitabine remains the only standard of care for this disease. More and more combination therapies containing gemcitabine have been tested or undergoing investigation. The interest in treating pancreatic cancer is apparently global. Over 75 abstracts were presented in the 2009 ASCO Gastrointestinal Cancers Symposium at San Francisco in the field of pancreatic cancer. In this highlights article, authors summarize the critical studies in the management of pancreatic cancer. A large retrospective study evaluated the role of post-operative adjuvant radiation (Abstract #181) and correlated the receipt of radiation with survival benefit. Borderline resectable pancreatic cancer remains an area that requires multi-disciplinary approach. Neo-adjuvant therapy very likely plays a role to downstage to a resectable state in these subgroup patients (Abstracts #197 and #248). In advanced or metastatic setting, studies aiming at the gemcitabine-based triplet or doublet combinations are still the mainstream. FFCD 0301 trial (Abstract #180), the only large phase III trial presented in the first-line setting, failed to demonstrate any survival advantage of either 5-FU and leucovorin plus cisplatin followed by gemcitabine or vice versa. Biologic agents containing regimens were also presented. Of note, gemcitabine and oxaliplatin plus bevacizumab achieved a high response rate of 39% (Abstract #182) while gemcitabine with dual monoclonal antibody regimen was disappointing (Abstract #183). The clear benefit of all other combinations over gemcitabine alone remains questionable given most studies are small. Newer agents, especially S-1 (Abstracts #213 and #251), are very promising, and further studies are warranted. In a nut shell, pancreatic cancer continues to pose an enormous challenge to clinicians and cancer scientists. With a more affluent world the global incidence of pancreatic cancer is rising. This meeting again emphasizes us that it is urgent to make big inroads into what still remains the most lethal of the common. PMID- 19287103 TI - Therapeutic tools in pancreatic cancer. AB - Pancreatic cancer is the fourth leading cause of cancer death in the United States and has a lower survival rate than other digestive tract tumors. It remains a therapeutic challenge with limited active agents. Honing our current understanding of markers of toxicity and response, and individualizing treatment with the prognostic and therapeutic tools available are important to make a worthy impact on a patient's course. The authors summarize selected abstracts from the ASCO Gastrointestinal Cancers Symposium, San Francisco, CA, USA, January 15-17, 2009. The Symposium featured pancreatic cancer in 84 research abstracts, of which, seven are reviewed that focus on markers of toxicity: cytidine deaminase (Abstract #151) and haptogloin (Abstract #167) as markers of gemcitabine toxicity; markers of response: use of PET scan for prognosis (Abstract #157), and correlations with CA 19-9 to post-chemo-radiation resectability (Abstract #215) and time to progression (Abstract #160); and individualized applications: characterizing the phenotypic similarities between a patient tumor and the direct xenograft (Abstract #154) and a report about the poor outcome of patients with ascites (Abstract #220). Validated clinical tools that can assist in managing patients through the narrow therapeutic window are needed. PMID- 19287104 TI - Early clinical experience using high intensity focused ultrasound for palliation of inoperable pancreatic cancer. AB - OBJECTIVE: To evaluate the safety and efficacy of high intensity focused ultrasound for palliation of inoperable pancreatic cancer in humans. PATIENTS: Eighty-nine patients with advanced pancreatic cancer were treated with high intensity focused ultrasound. There were 4 patients with stage II, 39 patients with stage III, and 46 patients with stage IV disease. The location of the tumors was as follows: head of pancreas 34 patients, body and/or tail of pancreas 55 patients. METHODS: Pain relief, local tumor control rate, median survival and complications were observed after high intensity focused ultrasound treatment. RESULTS: In the clinical treatments in humans the following local tumor control was seen: complete response, 0%; partial response, 14.6%; no change, 57.3%; progressive disease, 28.1%. Pain relief was achieved in 80.6% of patients who had pain prior to high intensity focused ultrasound therapy. The median survival was 26.0 months for patients with stage II disease, 11.2 months for patients with stage III disease, and 5.4 months for patients with stage IV disease. One-year survival rate was as follows: stage II, 100%; stage III, 41.0%; and stage IV, 6.5%. Two-year survival rate was as follows: stage II, 75.0%; stage III, 10.3%; and stage IV, 0%. Complications included superficial skin burns (3.4%), subcutaneous fat sclerosis (6.7%), and an asymptomatic pancreatic pseudocyst (1.1%). There were no severe complications or adverse events related to high intensity focused ultrasound therapy seen in any of the patients treated. CONCLUSIONS: Although this retrospective study has significant limitations, preliminary results suggest that the clinical application of high intensity focused ultrasound for pancreatic cancer appears to be safe and is a promising modality of treatment for palliation of pain related to pancreatic cancer. PMID- 19287105 TI - The proteome of mesenteric lymph during acute pancreatitis and implications for treatment. AB - CONTEXT: The protein fraction of mesenteric lymph during acute pancreatitis and other critical illness is thought to contain toxic factors. However, we do not have a complete description of the mesenteric lymph proteome during acute pancreatitis. OBJECTIVE: The aim of this study was to define the proteomic changes in mesenteric lymph during acute pancreatitis. SETTING: Animal Laboratory, University of Auckland, New Zealand. DESIGN: Mesenteric lymph was collected from sixteen male Wistar rats randomised to Group 1 (n=8) with taurocholate induced acute pancreatitis and Group 2 (n=8) sham control. The lymph was subjected to proteomic analysis using iTRAQ (Applied Biosystems, Foster City, CA, USA) and liquid chromatography-tandem mass spectrometry. RESULTS: Two hundred and forty-five proteins including 35 hypothetical proteins were identified in mesenteric lymph. Eight of the 245 proteins had a significant increase in their relative abundance in acute pancreatitis conditioned mesenteric lymph, and 7 of these were pancreatic catabolic enzymes (pancreatic amylase 2, pancreatic lipase, carboxypeptidase A2, chymotrypsinogen B, carboxypeptidase B1, cationic trypsinogen, ribonuclease 1). CONCLUSIONS: This is the first comprehensive description of the proteome of mesenteric lymph during acute pancreatitis and has demonstrated a significantly increased relative abundance of 7 secreted pancreatic catabolic enzymes in acute pancreatitis conditioned mesenteric lymph. This study provides a clear rationale for further research to investigate the efficacy of enteral protease inhibitors in the treatment of acute pancreatitis. PMID- 19287106 TI - Does endoscopic ultrasound have anything to offer in the diagnosis of idiopathic acute pancreatitis? AB - OBJECTIVE: The aim of this study was to investigate the role of EUS where other investigative techniques had failed to identify the cause of biochemically proven acute pancreatitis. SETTING: All biliary EUS examinations performed between January 2000 and December 2004 were identified from the radiology computerised database. PATIENTS: Forty-two patients (25 male, 17 female; mean age: 53+/-3.2 years) with negative prior radiological investigations underwent EUS. MAIN OUTCOME MEASURES: Prior and later radiological investigations, hospital readmission, and the need for further surgical intervention were also analysed. RESULTS: EUS was normal in 17 patients (40.5%) and demonstrated signs of recent acute pancreatitis but no other aetiological factor in 8 patients (19.0%). Cholelithiasis or microlithiasis was identified in 9 patients (21.4%), combined gallstones/microlithiasis and choledocholithiasis in was seen in 6 patients (14.3%). In one patient (2.4%), calculi were seen in the common bile duct but not the gallbladder. In a further case with recurrent acute pancreatitis (2.4%), chronic pancreatitis was diagnosed on EUS. All patients with common bile duct stones underwent ERCP and sphincterotomy, and stones were universally confirmed. One patient with gallbladder calculi alone required an ERCP after developing jaundice whilst awaiting cholecystectomy. CONCLUSIONS: EUS provided additional diagnostic information in 17 of the 42 patients (40.5%). Moreover, exclusion of gallstones/microlithiais is also important as it facilitates a search for other causes of pancreatitis. In conclusion, most cases of cholelithiasis can be diagnosed with standard imaging modalities but when these fail to identify a cause, EUS has an important role to play. PMID- 19287107 TI - A brief evaluation for pre-estimating the severity of gallstone pancreatitis. AB - CONTEXT: An uncomplicated and easy-to-use method of pre-estimating the severity of gallstone pancreatitis shortly after admission was required in order to direct clinicians for monitoring and/or transferring to a specialized center. OBJECTIVE: To determine the role of brief assessment in pre-estimating the severity of gallstone pancreatitis at admission. PATIENTS: Fifty-eight patients with gallstone pancreatitis were consecutively followed regarding the course of complications. MAIN OUTCOME VARIABLES: Sensitivity analysis of the Biliary Ranson score (>3), the modified Imrie score (>3), the acute physiology and chronic health evaluation II (APACHE-II) score (>5), white blood cell count (>14.5 x10(3)/dL), blood urea nitrogen (>12 mg/dL), random blood sugar (>150 mg/dL), pulse rate (>100 beats/min) and combinations of the four brief assessments were compared using the z-test. Two-tailed P values less than 0.05 were considered statistically significant. RESULTS: Both the Biliary Ranson score >3 and the modified Imrie score >3 had a sensitivity of 96.4% and a specificity of 96.7%. Both the APACHE-II score >5 and random blood sugar >150 mg/dL had a sensitivity of 89.3% which is comparable to a Biliary Ranson score >3 and a modified Imrie score >3. CONCLUSION: As compared to a Biliary Ranson score >3, a modified Imrie score >3 and an APACHE-II score >5, random blood sugar >150 mg/dL can be considered as an oversimplified and effective prognostic indicator at admission in patients with gallstone pancreatitis. PMID- 19287108 TI - The cyan fluorescent protein (CFP) transgenic mouse as a model for imaging pancreatic exocrine cells. AB - CONTEXT: The use of fluorescent proteins for in vivo imaging has opened many new areas of research. Among the important advances in the field have been the development of transgenic mice expressing various fluorescent proteins. OBJECTIVE: To report whole-body and organ-specific fluorescence imaging to characterize the transgenic cyan fluorescent protein mouse. DESIGN: Mice were imaged using two devices. Brightfield images were obtained with the OV100 Small Animal Imaging System (Olympus Corp., Tokyo, Japan). Fluorescence imaging was performed under the cyan fluorescent protein filter using the iBox Small Animal Imaging System (UVP, Upland, CA, USA). INTERVENTION: All animals were sacrificed immediately before imaging. They were imaged before and throughout multiple steps of a complete necropsy. Harvested organs were also imaged with both devices. Selected organs were then frozen and processed for histology, fluorescence microscopy, and H&E staining. Fluorescence microscopy was performed with an Olympus IMT-2 inverted fluorescence microscope. MAIN OUTCOME MEASURE: Determination of fluorescence intensity of different organs. RESULTS: Surprisingly, we found that there is differential enhancement of fluorescence among organs; most notably, the pancreas stands out from the rest of the gastrointestinal tract, displaying the strongest fluorescence of all organs in the mouse. Fluorescence microscopy demonstrated that the cyan fluorescent protein fluorescence resided in the acinar cells of the pancreas and not the islet cells. CONCLUSIONS: The cyan fluorescent protein mouse should lead to a deeper understanding of pancreatic function and pathology, including cancer. PMID- 19287109 TI - Enteral nutrition in severe acute pancreatitis. AB - CONTEXT: There is controversy concerning the merits of enteral and parenteral nutrition in the management of patients with severe acute pancreatitis. OBJECTIVE: This study was undertaken to evaluate the effect of enteral nutrition versus parenteral nutrition on serum markers of inflammation and outcome in patients with severe acute pancreatitis. SETTING: Tertiary care centre in North India. DESIGN: A prospective clinical trial. METHODS: Fifty consecutive patients with severe acute pancreatitis were randomized in a prospective trial to receive total enteral nutrition (n=25) or total parenteral nutrition (n=25). Enteral nutrition was delivered distal to the ligament of Treitz. Serum C-reactive protein, transferrin levels, albumin, surgical intervention, infections, duration of hospital stay and mortality were compared in the two groups. RESULTS: The mean age in the enteral nutrition group was 38.4+/-13.8 years and in the total parenteral nutrition group 41.1+/-11.3 years. The etiological factors were alcohol (n=19), gallstones (n=23), idiopathic (n=7) and drug-induced (n=1). There was a significant decrease in serum C-reactive protein values in both the enteral nutrition group and the total parenteral nutrition group at one week and two weeks (P<0.001 for both). Serum albumin rose from a prenutritional value of 2.82+/-0.51 g/dL to 3.34+/-0.45 g/dL on day 14 of nutritional support in the enteral nutrition group (P=0.003); in the total parenteral nutrition group, the level rose from 3.10+/-0.59 g/dL to 3.21+/-0.30 g/dL (P=0.638). A significant rise in transferrin value was observed from day 0 to day 14 in enteral nutrition group (169+/-30 to 196+/-36 mg/dL; P<0.001) whereas, in the total parenteral nutrition group, a less significant difference (191+/-41 to 201+/-29 mg/dL; P=0.044) was observed. There was no significant difference in surgical intervention (56.0% versus 60.0%; P=1.000), infective complications (64.0% versus 60.0%; P=1.000), hospital stay (42 days, 15-108 days, versus 36 days, 20-77 days; median, range; P=0.755), or mortality (20.0% versus 16.0%; P=1.000) in enteral nutrition versus total parenteral nutrition, respectively. CONCLUSION: Enteral nutrition and total parenteral nutrition are comparable in the management of severe acute pancreatitis in terms of hospital stay, need for surgical intervention, infections and mortality. PMID- 19287110 TI - A comparative analysis of pancreas cyst fluid CEA and histology with DNA mutational analysis in the detection of mucin producing or malignant cysts. AB - CONTEXT: The main objective of pancreatic cyst fluid analysis is to differentiate mucin-producing or malignant cysts from other cysts which have a benign course. K ras-2 point mutation and at least two mutations of allelic imbalance or loss of heterozygosity with good quality DNA has been suggested to predict mucinous cystic neoplasm (MCN). Elevated carcinogenic embryonic antigen (CEA) level in cyst fluid has also been shown to be predictive of mucinous or malignant cysts. OBJECTIVE: Identify the clinical impact of DNA mutational analysis of pancreatic cyst fluid with its correlation to cyst fluid chemistry and histologic analysis. PATIENTS: This retrospective analysis included all consecutive patients with pancreatic cysts who presented for evaluation by endoscopic ultrasound (EUS) with fine needle aspiration (FNA) over an 18 month period until November 2007. MAIN OUTCOME MEASURES: DNA analysis performed by Pathfinder TG (RedPath Integrated Pathology, Inc., Pittsburgh, PA, USA) and fluid CEA exceeding 192 ng/dL were used to suggest mucinous or malignant cysts. These parameters were compared to surgical histology or cytopathology of FNA specimens. RESULTS: Twenty-seven consecutive patients with cysts had samples submitted for DNA analysis which included 15 men and 12 women (mean age 62.8 and 61.3 years, respectively). In 20 patients, all parameters including cyst fluid, DNA analysis, and histology were available for comparison. Consistent findings were seen in 7/20 (35%) in which all parameters suggested negative benign findings. CEA level was elevated in 7 patients of which 4 had mucinous or malignant histology. In the remaining 13 patients with low CEA levels, 11 had negative histology. The sensitivity and specificity of CEA based on these results was 66% and 78.6% respectively. The positive predictive value (PPV) of CEA was 57% and the negative predictive value (NPV) was 84.6%. K-ras-2 mutation was detected in 3 patients, absent in 17 patients and falsely negative in 4 cases based on histology. The sensitivity and specificity were 33% and 92.6% respectively. The PPV was 66% and NPV was 76%. Detection of loss of heterozygosity mutations was noted in 7 patients, of which 4 were falsely positive. In the remaining 13 patients, 3 were falsely negative. The sensitivity and specificity were 50% and 71% respectively. The PPV was 42.9% and NPV was 76.9%. In a group of 6 patients with available surgical histology demonstrating mucin-producing or malignant cysts, fluid CEA level had a sensitivity of 66.7%. However, K-ras-2 and loss of heterozygosity mutational analysis had a much lower sensitivity at 33% and 50% respectively. CONCLUSIONS: Consistency in histology, CEA levels, and K-ras-2 and loss of heterozygosity mutations was seen in only 35% of cases, all of which were benign cysts. In the detection of malignant cysts, elevated CEA levels were more predictive of histology in comparison to K-ras-2 or loss of heterozygosity mutations. Additionally, false positivity of loss of heterozygosity mutations was noted to be considerably higher than K-ras-2 mutations or even fluid CEA levels. These findings suggest that DNA mutation analysis should not be used routinely but rather selectively in the evaluation of pancreatic cysts. PMID- 19287111 TI - Pancreaticoduodenectomy in patients with a history of Roux-en Y gastric bypass surgery. AB - CONTEXT: Roux-en Y gastric bypass surgery is the most common operation for treatment of morbid obesity. The approach to pancreaticoduodenal resection in patients with a history of Roux-en Y gastric bypass is not well described. CASE REPORTS: Pancreaticoduodenal resection was performed in two patients with distal bile duct strictures, with a past history of Roux-en Y gastric bypass. In both cases the remnant stomach, distal bile duct, duodenum and pancreas were excised. The biliopancreatic limb was divided close to the ligament of Treitz and brought up into the supracolic compartment in a retromesenteric manner and pancreatic and biliary anastomoses performed. The previous enteroenterostomy and gastrojejunal anastomoses were left intact. Both patients had an uncompleted post-operative recovery. The mean operating time was 6.5 hours and mean estimated blood loss was 525 mL. They were discharged home by days 6 and 7 post-operatively. CONCLUSIONS: Pancreaticoduodenal resection can be successfully performed following Roux-en Y gastric bypass with en-bloc excision of the remnant stomach, with the pancreas and bile duct anastomosed to the divided biliopancreatic limb. PMID- 19287112 TI - Disparities in demographics among patients with pancreatitis-related mortality. AB - CONTEXT: Hospital admissions for pancreatitis are increasing. Factors involved in inpatient mortality have not been previously assessed on a large-scale basis. OBJECTIVE: The aim was to study factors associated with pancreatitis-related death in hospitalized patients. SETTING: Retrospective analysis of the 2004 U.S. Healthcare Cost and Utilization Project (HCUP) database was performed using "pancreatitis" as admitting diagnosis and "mortality" as primary endpoint. MAIN OUTCOME MEASURES: Age, race, gender, income, length of stay, number of diagnoses, and number of procedures were identified as candidate risk factors associated with death. DESIGN: Univariate and multivariate logistic regression analyses were performed to identify significant covariates. RESULTS: In 2004, total of 78,864 patients were admitted with pancreatitis; 2,129 (2.7%) patients died. Complete data were available for 57,068 patients. Age greater than 65 was 3 times more often associated with mortality (OR=2.92; P<0.001), while females were 19% less likely to die (OR=0.81; P<0.001). African American patients were 18% more likely to die than whites (OR=1.18, P=0.025), and increasing length of stay was associated with increasing mortality (more than 14 days compared with less than 3 days: OR=1.24; P=0.004). Patients with more than 3 diagnoses and more than one hospital procedure were 17 times (OR=16.7; P<0.001) and 5 times (OR=5.42; P<0.001) more likely to die, respectively. Compared to the lowest income quartile, patients in the 2nd and 3rd quartiles were 19% (OR=0.81; P=0.004) and 17% (OR=0.83; P=0.016) less likely to die, respectively. CONCLUSION: Age greater than 65 years, male gender, multiple diagnoses, African American race and low income are strongly associated with inpatient mortality from pancreatitis. Increased number of procedures and longer length of stay are also highly correlative with death. PMID- 19287113 TI - Endoscopic diagnosis of squamous cell carcinoma of the pancreas invading the stomach. AB - CONTEXT: Squamous cell carcinoma of the pancreas is an unusual tumor due to the absence of squamous cells in the normal pancreas. Its clinical presentation is similar to that of adenocarcinoma of the pancreas and is usually diagnosed intraoperatively or at autopsy. CASE REPORT: We report a 50-year-old male who presented with obstructive jaundice and symptoms suggestive of gastric outlet obstruction. Contrast-enhanced computerized tomography revealed a heterogeneous enhancing mass in the head of pancreas and upper gastrointestinal endoscopy revealed an ulcerated polypoidal lesion in the stomach; the endoscopic biopsies taken from this region revealed infiltration of the lamina propria with malignant squamous cells. CONCLUSION: A rare case of squamous cell carcinoma of the head of the pancreas presenting with obstructive jaundice and gastric outlet obstruction and in which diagnosis was established by endoscopic biopsies of the stomach has been described. PMID- 19287114 TI - Isolated tuberculosis of the ampulla of vater masquerading as periampullary carcinoma: a case report. AB - CONTEXT: Isolated tuberculosis of the ampulla of Vater has not yet been reported. The clinical features of isolated periampullary tuberculosis are at times similar to those seen in patients with periampullary carcinoma. Diagnosis is difficult, and biopsy and culture of the suspected lesion are often negative for Mycobacterium tuberculosis. CASE REPORT: We herein describe one such case masquerading as periampullary carcinoma in a 70-year-old woman. Due to comorbid conditions only a local excision of the ampulla was carried out. Histopathology revealed giant cells in the absence of caseation necrosis and the presence of Mycobacterium tuberculosis was proven using the polymerase chain reaction. CONCLUSION: Isolated tuberculosis of ampulla of Vater is extremely rare but must be kept in mind when making the differential diagnosis of isolated ampullary lesion. PMID- 19287115 TI - Pancreatic plasmacytoma presenting as variceal hemorrhage: life threatening complication of a rare entity. AB - CONTEXT: Pancreatic plasmacytoma is a rare entity and presents with features of mass lesion of pancreas. CASE REPORT: We present an interesting case of pancreatic plasmacytoma with life threatening gastrointestinal bleeding secondary to isolated gastric varices. CONCLUSION: This case highlights the importance of considering it in differential diagnosis of patients with anemia, recurrent pancreatitis or jaundice and isolated gastric varices, prompting a CT scan to evaluate for any pancreatic mass lesions. PMID- 19287116 TI - Extra-gastrointestinal stromal tumor presenting as hemorrhagic pancreatic cyst diagnosed by EUS-FNA. AB - CONTEXT: Pancreatic gastrointestinal stromal tumors (GIST) are rare mesenchymal tumor with only 6 cases reported to date. We report a case of pancreatic GIST presenting as hemorrhagic cyst. CASE REPORT: A 63-year-old female with past medical history of hypertension and pancreatic mass presented with fatigue. She was found to have anemia requiring blood transfusion. An abdominal CT scan revealed an 11x16 cm cystic mass at the pancreatic body that had increased in size compared with previous CT scan. Endoscopic ultrasound confirmed a large complex pancreatic mass and fine needle aspiration demonstrated gross bloody fluid. Cytology revealed a spindle cell lesion. Immunohistochemistry from cyst wall biopsy was strongly positive for CD34 and CD117 confirming the diagnosis of pancreatic GIST. CONCLUSION: We report a case of pancreatic GIST which presented as hemorrhagic cyst. Endoscopic ultrasound guided fine needle aspiration plays an important role in the diagnosis. Although it is an uncommon tumor, pancreatic GIST should be in differential diagnosis of hemorrhagic pancreatic cystic lesions as well as a rare cause of solid pancreatic lesions. PMID- 19287117 TI - Pancreatoblastoma in an adult patient. A case report. AB - CONTEXT: Preoperative diagnosis of a pancreatoblastoma in adults is challenging because of its rarity. Furthermore, difficulties increase since pancreatoblastomas share radiological findings similar to those found in other masses of the pancreas. CASE REPORT: A 36-year-old woman was studied with ultrasonography and CT for a mass of the pancreatic head causing obstructive jaundice. Diagnosis of pancreatoblastoma was obtained with histology and immunohistochemistry of the resected specimen. CONCLUSION: We reviewed the radiological findings of pancreatoblastomas and possible radiological criteria of differentiation from other pancreatic tumors. A pancreatoblastoma should be considered in the differential diagnosis of a pancreatic mass presenting atypical radiological features. PMID- 19287118 TI - Pancreas sparing resection for giant hamartoma of Brunner's glands. AB - CONTEXT: Benign proliferative changes of the Brunner's glands account for about 10% of neoplasias of the duodenal bulb. CASE REPORT: The authors present a case of giant hamartoma of Brunner's glands. The patient presented clinically with vague epigastric discomfort. A pancreas-sparing duodenal resection was performed. The patient made an uneventful recovery and is well at a one year follow-up. The diagnosis was established on histopathology. CONCLUSION: Duodenal hamartomas are rare and may become very large, mandating surgical removal. A pancreas-sparing duodenal resection is one of the modalities of treating such lesions. PMID- 19287119 TI - Pancreatic lipoma diagnosed using EUS-FNA. A case report. AB - CONTEXT: Pancreatic mesenchymal tumors are rare, accounting for only 1-2% of pancreatic tumors. A pancreatic lipoma is an especially rare condition. This is only the second report of a pancreatic lipoma diagnosed before surgery using endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNA). CASE REPORT: A 75-year-old woman with a round low-density lesion which had been detected by CT was referred to our hospital. EUS revealed an oval 49x32 mm homogeneous and almost isoechoic mass (as compared to normal pancreatic parenchyma) in the pancreatic head. Its outer margin was not clearly differentiated from the parenchyma. Although a benign pancreatic lipomatous lesion was deemed most probable from information obtained using CT/MRI, a mass with malignancy such as a liposarcoma could not be ruled out as a differential diagnosis. Therefore, EUS-FNA was performed for a definitive diagnosis. Two passes were performed with on-site pathology. The results obtained from the sample analysis indicated a pancreatic lipoma consisting of mature fat cells with no atypia. CONCLUSION: Although imaging modalities are useful for the diagnosis of pancreatic lipomatous lesions, it is sometimes difficult to diagnose lipomatous malignancies mimicking benign lipoma. This case demonstrates the usefulness of EUS-FNA for the differential diagnosis of pancreatic lipomatous lesions. PMID- 19287120 TI - An unusual mixed tumor of the pancreas: sonographic and MDCT features. AB - CONTEXT: Mixed tumors of the pancreas are exceedingly rare. CASE REPORT: We herein report on a 54-year-old female who presented with an enlarging cystic lesion in the head of the pancreas. Right upper quadrant ultrasound and multidetector-row CT scan showed a well-defined unilocular cystic tumor located in the head of the pancreas and surrounded, in part, by a hypervascular solid mass. CONCLUSION: Although mixed exocrine/endocrine pancreatic tumors have been described previously, to the best of our knowledge, this is the first case of a pancreatic mixed intraductal papillary mucinous neoplasm/endocrine tumor with illustration of its ultrasound and CT features. Moreover, the importance of preoperative analysis of imaging features in the assessment of pancreatic neoplasms is discussed. PMID- 19287121 TI - Frantz's tumor (solid pseudopapillary tumor) of the pancreas. A case report. AB - CONTEXT: A solid pseudopapillary tumor of the pancreas is a rare neoplasm which, for the most part, affects young women and has a relatively favorable prognosis with a low malignant potential. These tumors usually have unclear clinical features and may form very large masses before being diagnosed. CASE REPORT: We report the case of a 29-year-old woman who underwent complete resection of the tumor using a distal pancreatectomy and splenectomy procedure. The patient is being followed-up and in good condition. A review of the relevant literature is also presented. CONCLUSIONS: A solid pseudopapillary tumor of the pancreas is a rare condition with a low potential for malignancy and favorable prognosis; surgical resection is generally curative. PMID- 19287122 TI - Oral osteonecrosis associated with the use of zoledronic acid: first case of a patient with advanced pancreatic cancer and bone metastases. PMID- 19287123 TI - Cardiotoxicity of 5-fluorouracil and capecitabine in a pancreatic cancer patient with a novel mutation in the dihydropyrimidine dehydrogenase gene. AB - CONTEXT: 5-fluorouracil (5-FU) is an antimetabolite that acts during the S phase of the cell cycle. Dihydropyrimidine dehydrogenase (DPD) is the initial and rate limiting enzyme in the pathway that catabolises the pyrimidines. 5-fluorouracil and its oral prodrug capecitabine are used in the treatment of a number of solid tumors, including colorectal, breast, gastric, pancreatic, prostate, and bladder cancers. Common side effects include leukopenia, diarrhea, stomatitis, nausea, vomiting, and alopecia. Cardiotoxicity is a relatively uncommon side effect of 5 fluorouracil and capecitabine. CASE REPORT: This article reports the case of a 63 year-old male with locally invasive pancreatic cancer who developed recurrent chest pain and ischemic electrocardiogram changes after treatment with 5 fluorouracil and capecitabine. Full sequencing of the dihydropyrimidine dehydrogenase (DPYD) gene and analysis of the thymidylate synthetase (TYMS) gene promoter region was performed. Pharmacogenetic testing revealed p453L (1358C>T) type DPYD germ line mutation. This mutation has not been reported previously in association with 5-fluorouracil induced cardiotoxicity. CONCLUSION: Cardiotoxicity associated with 5-fluorouracil and capecitabine administration is infrequently reported in the literature and appears to be dose and schedule dependent. Genetic variations such as polymorphic abnormality of DPYD are potential causative factors for a significant portion of serious adverse reactions to 5-fluorouracil-based therapy. PMID- 19287124 TI - Ischemic acute necrotizing pancreatitis in a marathon runner. Comment. PMID- 19287126 TI - Antibiotic prophylaxis in severe acute pancreatitis: do we need more meta analytic studies? PMID- 19287127 TI - Genipin cross-linked fibrin hydrogels for in vitro human articular cartilage tissue-engineered regeneration. AB - Our objective was to examine the potential of a genipin cross-linked human fibrin hydrogel system as a scaffold for articular cartilage tissue engineering. Human articular chondrocytes were incorporated into modified human fibrin gels and evaluated for mechanical properties, cell viability, gene expression, extracellular matrix production and subcutaneous biodegradation. Genipin, a naturally occurring compound used in the treatment of inflammation, was used as a cross-linker. Genipin cross-linking did not significantly affect cell viability, but significantly increased the dynamic compression and shear moduli of the hydrogel. The ratio of the change in collagen II versus collagen I expression increased more than 8-fold over 5 weeks as detected with real-time RT-PCR. Accumulation of collagen II and aggrecan in hydrogel extracellular matrix was observed after 5 weeks in cell culture. Overall, our results indicate that genipin appeared to inhibit the inflammatory reaction observed 3 weeks after subcutaneous implantation of the fibrin into rats. Therefore, genipin cross linked fibrin hydrogels can be used as cell-compatible tissue engineering scaffolds for articular cartilage regeneration, for utility in autologous treatments that eliminate the risk of tissue rejection and viral infection. PMID- 19287128 TI - Cartilage collagen fibril network in newborn transgenic mice analyzed by electron microscopic stereology. AB - The objective was to investigate by electron microscopic stereology the properties of the cartilage collagen fibril network in newborn transgenic mice. The mice harbored transgenes targeted to affect the structure or assembly of the collagen fibrils. The mouse lines investigated here harbored either (i) one or (ii) two human COL2A1 genes with Arg519Cys mutation in addition to one or (iii) no active allele(s) of the murine COL2A1 gene, (iv) two inactive alleles of the procollagen N-proteinase genes, or (v) a human COL2A1 gene with deleted exons 16 27. In all newborn mice carrying the COL2A1 transgene with Arg519Cys mutation, the growth plate collagen fibrils were thinner than in the wild-type (wt) mice and showed clearly reduced volume fraction of the fibril network. In mice with the inactive procollagen N-proteinase genes, the fibril thickness and the volume fraction of collagen did not differ from the wt mice. In mice harboring the transgene of human COL2A1 gene with internally deleted exons 16-27, the collagen fibril diameter remained the same, but the volume density of collagen network was reduced. Using the indirect stereology, the differences in the collagen fibril stereological estimates could be reliably detected in newborn mice harboring mutations that affect the structure and assembly of collagen fibrils. The EM stereology permitted early detection of altered phenotype of the collagen fibril network in newborn transgenic mice. It is recommended that the indirect model based stereological technique is utilized instead of the direct design-based technique for the estimation of collagen volume, surface, and length densities. PMID- 19287129 TI - Efferent projections of the anterior and posterodorsal regions of the medial nucleus of the amygdala in the mouse. AB - The efferent projections of the anterior and posterodorsal part of the medial nucleus (MePD) in the mouse were studied by means of anterograde axonal tracing using biotinylated dextran amine. The MePD axons ran mainly via the stria terminalis and to a lesser extent via the ventral amygdalofugal pathway. The projections to the forebrain were broadly distributed and varied from very strong to scant. The most significant connections were destined to the bed nucleus of the stria terminalis in which all parts of the medial division were innervated by MePD neurons. Moderate projections reached the limbic striatum (nucleus accumbens), olfactory tubercle and the lateral septal nucleus. The substantia innominata was also innervated by the MePD, and especially the projection to its ventral portion was substantial. The profuse innervation of the medial preoptic nucleus and medial preoptic area indicated significant involvement of the MePD in sexual behavior. Many hypothalamic nuclei were innervated but to a different extent. The very strong innervation of the ventral premammillary nucleus further indicated the involvement of the MePD in the neuronal circuitry for sexual behavior. Substantial projections also reached the anterior hypothalamus and tuber cinereum, while the connections to the lateral hypothalamus were widespread but showed moderate density. MePD strongly innervated the ventrolateral part of the ventromedial hypothalamic nucleus and moderately its remaining parts. The neurosecretory hypothalamic nuclei and the arcuate nucleus contained only a few MePD terminals. The thalamic innervation was very scant and reached the lateral habenular nucleus and the nuclei of the midline. The mesencephalic connections were moderate to sparse and projected to the mesolimbic dopaminergic groups in the ventral tegmental area, the pars lateralis and the dorsal tier of the substantia nigra pars compacta, the periaqueductal gray and the dorsal raphe nucleus. The present results principally resembled data known in other rodent species; however, the efferents of the MePD often differed in extent and/or topical distribution. PMID- 19287130 TI - Effect of severe aortic stenosis on the outcome in elderly patients undergoing repair of hip fracture. AB - BACKGROUND: The perioperative assessment and management of elderly patients with hip fracture and significant aortic stenosis (AS) is an increasingly common clinical problem with little data available to guide perioperative management. OBJECTIVES: It was the aim of this study to examine the incidence of perioperative events in an elderly population of patients with severe AS undergoing repair of hip fracture as compared with controls without severe AS. METHODS: Patients over the age of 70 with an echocardiographic diagnosis of severe AS defined as an aortic valve area /=2 independent control studies) amongst 221 subjects with an autism spectrum disorder (ASD) and/or intellectual disability (ID) in comparison to 40 controls using three array platforms of increasing resolution (Spectral Genomics (1 Mb), Agilent (0.03 Mb) and NimbleGen (0.01 Mb)). We determined that the number of bCNVs/subject, type and frequency of most common bCNVs were similar for both the test and control cohorts when the same array platform was used. The comparison of the 'load' of bCNVs (i.e. number/subject) to a standardized metric of phenotypic features (see de Vries et al., 2001) in 91 ASD subjects revealed that a phenotype score >/=4 is significantly more common (P < 0.05) in persons with an ASD having one or more bCNVs via 1 Mb array-CGH, whereas individuals without any recognizable bCNVs are significantly more likely to have a less complex phenotype and a score 60 years old) people as compared to healthy young (<45 years old) people in the frequency of the individuals with a low copy number of the C4B gene (one of the two genes (C4A and C4B) encoding the C4 complement protein). This observation indicated that subjects with a low C4B copy number are selected out from the population of healthy individuals due to their increased morbidity/mortality of some disease(s). In accordance with this assumption a marked accumulation of low C4B copy number carriers was found in patients with acute myocardial infarction (AMI) and stroke as compared to age-matched healthy subjects. In addition it was demonstrated that AMI patients who carry a low copy number of the C4B gene have a highly significantly increased risk for short-term (1 year) post-AMI mortality as compared to non-carriers. These relationships, which were first revealed more than 15 years ago by our group, have been recently supported by new data obtained using up-to-date genotyping methods worked out by our group in two Caucasian populations, Hungarian and Icelandic. In addition, we proved that this relationship concerns only recent smokers. These findings indicate that there is strong connection between low copy number of the C4B gene on one hand and cardiovascular disease morbidity and mortality on the other. Hypotheses and the results of recent studies aiming to elucidate the mechanism of this association as well as other diseases connected with the low C4B copy number are discussed. PMID- 19287147 TI - Phenotypes, genotypes and disease susceptibility associated with gene copy number variations: complement C4 CNVs in European American healthy subjects and those with systemic lupus erythematosus. AB - A new paradigm in human genetics is high frequencies of inter-individual variations in copy numbers of specific genomic DNA segments. Such common copy number variation (CNV) loci often contain genes engaged in host-environment interaction including those involved in immune effector functions. DNA sequences within a CNV locus often share a high degree of identity but beneficial or deleterious polymorphic variants are present among different individuals. Thus, common gene CNVs can contribute, both qualitatively and quantitatively, to a spectrum of phenotypic variants. In this review we describe the phenotypic and genotypic diversities of complement C4 created by copy number variations of RCCX modules (RP-C4-CYP21-TNX) and size dichotomy of C4 genes. A direct outcome of C4 CNV is the generation of two classes of polymorphic proteins, C4A and C4B, with differential chemical reactivities towards peptide or carbohydrate antigens, and a range of C4 plasma protein concentrations (from 15 to 70 mg/dl) among healthy subjects. Deliberate molecular genetic studies enabled development of definitive techniques to determine exact patterns of RCCX modular variations, copy numbers of long and short C4A and C4B genes by Southern blot analyses or by real-time quantitative PCR. It is found that in healthy European Americans, the total C4 gene copy number per diploid genome ranges from 2 to 6: 60.8% of people with four copies of C4 genes, 27.2% with less than four copies, and 12% with more than four copies. Such a distribution is skewed towards the low copy number side in patients with systemic lupus erythematosus (SLE), a prototypic autoimmune disease with complex etiology. In SLE, the frequency of individuals with less than four copies of C4 is significantly increased (42.2%), while the frequency of those with more than four copies is decreased (6%). This decrease in total C4 gene copy number in SLE is due to increases in homozygous and heterozygous deficiencies of C4A but not C4B. Therefore, it is concluded that lower copy number of C4 is a risk factor for and higher gene copy number of C4 is a protective factor against SLE disease susceptibility. PMID- 19287149 TI - Copy number variation of beta-defensins and relevance to disease. AB - Beta-defensins are multifunctional proteins that have an important role in the innate immune system. A cluster of beta-defensin genes shows common and extensive variation in copy number in humans. This copy number has been associated with differing susceptibility to Crohn's disease and psoriasis. In this review, I summarise what is currently known about copy number variation (CNV) of beta defensins, and discuss practical considerations when studying these genes, and copy number variation in general. I also suggest future directions for research, with an emphasis on improvement of methods for accurately typing CNV. PMID- 19287148 TI - Copy number variants in genetic susceptibility and severity of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a systemic autoimmune disorder characterized by the presence of auto-antibodies to nuclear antigens, immune complex deposition, and subsequent tissue destruction. Early studies in twins suggested that SLE has, at least in part, a genetic basis, and a role for class II alleles in the major histocompatibility complex has been known for over 30 years. Through both linkage studies and candidate gene studies, numerous additional genetic risk factors have been identified. The recent publication of two SNP-based genome-wide association studies (GWAS) has resulted in the confirmation of a number of previously identified genetic risk loci and has identified new previously unappreciated loci conferring risk for development of SLE. A role for gene copy number variation (CNV) in SLE has also been appreciated through studies of the complement component 4 (C4) loci and more recent work in the IgG Fc receptor loci. The availability of large SNP-based GWAS datasets will undoubtedly lead to the genome-wide analysis and identification of copy number variants related to genetic susceptibility for development of SLE. We review current studies of CNV in SLE susceptibility that include reports of association between SLE and CNV in C4, IgG Fc receptors, TLR7, and CCL3L1. PMID- 19287150 TI - HIV-1/AIDS susceptibility and copy number variation in CCL3L1, a gene encoding a natural ligand for HIV-1 co-receptor CCR5. AB - Variations of gene copy number in the human genome are increasingly recognized as a genetic factor in phenotypic variation. Human CC chemokine ligand 3-like 1 gene (CCL3L1), which is located on human chromosome 17q11.2, is highly variable in copy number owing to having a hot spot for segmental duplications. CCL3L1, a natural ligand for HIV-1 co-receptor CCR5, is a potent HIV-1-suppressive chemokine. CCL3L1 copy number variation (CNV) is tightly linked to HIV-1/AIDS susceptibility, and a lower copy number is associated with an enhanced risk for acquiring HIV-1 and also progressing more rapidly to AIDS and death. In this article we review recent studies to evaluate the association between the CCL3L1 copy number and HIV-1/AIDS susceptibility. PMID- 19287151 TI - Copy number variation of Fc gamma receptor genes and disease predisposition. AB - Naturally occurring variation in gene copy number is increasingly recognized as a major source of inter-individual differences in phenotype and is an important susceptibility factor for genetically complex diseases. Several studies provide evidence of copy number variation at genes involved in inflammation and immunity highlighting their possible contribution to the inter-individual variation observed in immune responses. This review will explore copy number variation at the Fc gamma receptor cluster and its relevance in the pathogenesis of common human diseases. PMID- 19287152 TI - Copy number variation in metabolic phenotypes. AB - Despite successes in identifying genetic contributors to common metabolic phenotypes, only part of the heritable component of these traits has thus far been explained. Copy number variation (CNV) is likely to be responsible for some of the unexplained variation. As observed with single nucleotide changes, it is probable that both rare and common CNVs will contribute to susceptibility to metabolic disease. For instance, CNVs in the LDLR gene underlie a substantial portion of disease in patients with heterozygous familial hypercholesterolemia. As well, a common CNV in LPA encoding apolipoprotein(a) is the primary determinant of plasma lipoprotein(a) concentrations, a risk factor for atherosclerosis. Recent efforts to map CNVs in control populations have defined their size, frequency and distribution. Many of the identified CNVs overlap genes with important functions in metabolic pathways. The overlap of CNVs that were found in control datasets with functional candidate genes or genes with previous evidence of association with metabolic syndrome presents an important subset for future CNV association studies. Finally, we describe an approach to search for CNVs in a rare high-penetrance metabolic disorder, namely lipodystrophy. As methods to identify CNVs increase in precision and accuracy, the prospect of identifying their role in both rare Mendelian and common complex metabolic phenotypes will become a reality. PMID- 19287153 TI - Copy number alterations and copy number variation in cancer: close encounters of the bad kind. AB - Recent studies have unveiled copy number variants (CNVs) as an important source of genetic variation. Many of these CNVs contain coding sequences, which have been shown to be dosage sensitive. Evidence is accumulating that certain CNVs have impact on susceptibility to human diseases such as HIV infection and autoimmune diseases, as well as on adaptability to environmental conditions or nutrition. The possible role and impact of CNVs on cancer development and progression is only now emerging. In this review we look into the role of CNVs and their associated genomic structural features in relation to the formation of chromosome alterations in cancer cells and evolutionary genomic plasticity, as well as the de novo occurrence of known or putative CNVs as somatic events during oncogenesis. The role of germline CNVs in cancer predisposition is still largely unexplored. A number of observations seem to warrant the importance of further studies to elucidate the impact of these variants in the early steps of carcinogenesis. PMID- 19287154 TI - Chromosome copy number variation and breast cancer risk. AB - Breast cancer is the most frequent cancer among women. It is caused by genetic and environmental factors. Whereas mutations in high-penetrance susceptibility genes have been identified in familial breast cancer and several single nucleotide polymorphisms (SNPs) have been shown to be associated with both familial and sporadic breast cancer risk, the impact of genomic copy number variants (CNVs) on breast cancer risk has so far poorly been studied. An example of a CNV affecting the tumor suppressor gene MTUS1 that has been shown to be associated with familial breast cancer risk is given. Moreover, we discuss CNVs affecting detoxification genes like GSTM1 and GSTT1 whose association with breast cancer risk is controversial. Finally, the potential of array-based genome-wide CNV association studies is discussed. PMID- 19287155 TI - Predisposition to colorectal cancer: exploiting copy number variation to identify novel predisposing genes and mechanisms. AB - Although cancer is mostly regarded as an acquired disease, familial predisposition plays a significant role in many cancer types. Thus far, several high penetrant cancer predisposing genes have been identified. As yet, however, these genes explain only a fraction of the familial and/or hereditary cases of cancer. This has led to the exploration of the human genome for novel cancer predisposing genes. The identification of such genes will not only increase our understanding of cancer predisposition and development, but will also have direct implications for genetic counseling and personalized management of the patients and their family members. Here we provide an inventory of currently known molecular mechanisms related to familial colorectal cancer development and an outline of copy number analysis-based strategies to identify new predisposing genes. Finally, we discuss a novel copy number-associated epigenetic mechanism underlying the predisposition to colorectal cancer. PMID- 19287156 TI - CNVs of human genes and their implication in pharmacogenetics. AB - Pharmacogenetics encompasses genetic variation with importance for drug response and adverse drug reactions with emphasis on drug transporters, drug metabolizing enzymes, and drug receptors. The highest penetrance with respect to drug action is generally observed for variability in genes encoding drug metabolizing enzymes, and gene copy number variations play a very important role in this respect. Alleles containing 0-13 active gene copies have been described, and this variation affects the clinical outcome of treatment for about 20-30% of all drugs. Gene copy number variation has also an influence on nicotine metabolism and detoxification by glutathione transferases and sulfotransferases. In the current overview we provide an update of the situation with emphasis on clinically important examples. PMID- 19287157 TI - Sulfotransferase gene copy number variation: pharmacogenetics and function. AB - Pharmacogenetics is the study of the role of inheritance in variation to drug response. Drug response phenotypes can vary from adverse drug reactions at one end of the spectrum to equally serious lack of the desired effect of drug therapy at the other. Many of the current important examples of pharmacogenetics involve inherited variation in drug metabolism. Sulfate conjugation catalyzed by cytosolic sulfotransferase (SULT) enzymes, particularly SULT1A1, is a major pathway for drug metabolism in humans. Pharmacogenetic studies of SULT1A1 began over a quarter of a century ago and have advanced from biochemical genetic experiments to include cDNA and gene cloning, gene resequencing, and functional studies of the effects of single nucleotide polymorphisms (SNPs). SNP genotyping, in turn, led to the discovery of functionally important copy number variations (CNVs) in the SULT1A1 gene. This review will briefly describe the evolution of our understanding of SULT1A1 pharmacogenetics and CNV, as well as challenges involved in utilizing both SNP and CNV data in an attempt to predict SULT1A1 function. SULT1A1 represents one example of the potential importance of CNV for the evolving disciplines of pharmacogenetics and pharmacogenomics. PMID- 19287158 TI - Germline copy number variation in control populations. AB - Copy number variation (CNV) is an important source of genomic variation. Characterizing CNVs in phenotypically normal individuals is important for understanding the functional significance of these variants. Many studies have reported CNVs in control populations, but wide variability is observed in their design and outcome. Importantly, medical and phenotypic information for control populations must be carefully documented, and accurate genotyping will be necessary to determine the population genetics of CNVs. Despite existing challenges in studying this class of variants, it is evident that CNVs are ubiquitous in human genomes, with non-random distribution, and they affect thousands of coding regions, potentially contributing to human disease and phenotypic variability. Higher-resolution detection platforms and improved algorithms will further define our understanding of CNVs in control populations, leading to development of effective disease-association studies. PMID- 19287159 TI - Large-scale copy number variants (CNVs) detected in different ethnic human populations. AB - The large-scale copy-number variations (CNVs) in the human genome are associated with developmental disorders and susceptibility to diseases. More importantly, CNVs may represent a major genetic component of our phenotypic diversity. Following the development of methodologies and introduction of new research platforms, accumulation of the nature and pattern of CNVs from normal populations has progressed. The examination of relatively large numbers of specific ethnic groups has recently started. Although the results are not always consistent, it is likely that different human populations bear different CNVs, as is the case for single-nucleotide polymorphisms (SNPs) and insertion-deletion (indel) polymorphisms. We review recent publications about the nature of inter population, especially inter-ethnic group, differences of CNVs. PMID- 19287160 TI - Human copy number polymorphic genes. AB - Recent large-scale genomic studies within human populations have identified numerous genomic regions as copy number variant (CNV). As these CNV regions often overlap coding regions of the genome, large lists of potentially copy number polymorphic genes have been produced that are candidates for disease association. Most of the current data regarding normal genic variation, however, has been generated using BAC or SNP microarrays, which lack precision especially with respect to exons. To address this, we assessed 2,790 candidate CNV genes defined from available studies in nine well-characterized HapMap individuals by designing a customized oligonucleotide microarray targeted specifically to exons. Using exon array comparative genomic hybridization (aCGH), we detected 255 (9%) of the candidates as true CNVs including 134 with evidence of variation over the entire gene. Individuals differed in copy number from the control by an average of 100 gene loci. Both partial- and whole-gene CNVs were strongly associated with segmental duplications (55 and 71%, respectively) as well as regions of positive selection. We confirmed 37% of the whole-gene CNVs using the fosmid end sequence pair (ESP) structural variation map for these same individuals. If we modify the end sequence pair mapping strategy to include low-sequence identity ESPs (98 99.5%) and ESPs with an everted orientation, we can capture 82% of the missed genes leading to more complete ascertainment of structural variation within duplicated genes. Our results indicate that segmental duplications are the source of the majority of full-length copy number polymorphic genes, most of the variant genes are organized as tandem duplications, and a significant fraction of these genes will represent paralogs with levels of sequence diversity beyond thresholds of allelic variation. In addition, these data provide a targeted set of CNV genes enriched for regions likely to be associated with human phenotypic differences due to copy number changes and present a source of copy number responsive oligonucleotide probes for future association studies. PMID- 19287161 TI - Human subtelomeric copy number variations. AB - Copy number variation is a defining characteristic of human subtelomeres. Human subtelomeric segmental duplication regions ('Subtelomeric Repeats') comprise about 25% of the most distal 500 kb and 80% of the most distal 100 kb in human DNA. Huge allelic disparities seen in subtelomeric DNA sequence content and organization are postulated to have an impact on the dosage of transcripts embedded within the duplicated sequences, on the transcription of genes in adjacent single copy DNA regions, and on the chromatin structures mediating telomere functions including chromosome stability. In addition to the complex duplicon substructure and huge allelic variations in extended subtelomere regions, both copy number variation and alternative sequence organizations for DNA characterize the sequences immediately adjacent to terminal (TTAGGG)n tracts ('subterminal DNA'). The structural variation in subterminal DNA is likely to have important consequences for expression of subterminal transcripts such as a newly-discovered gene family encoding actin-interacting proteins and a non-coding telomeric repeat containing RNA (TERRA) transcript family critical for telomere integrity. Major immediate challenges include discovering the full extent and nature of subtelomeric structural and copy number variation in humans, and developing methods for tracking individual allelic variants in the context of total genomic DNA. PMID- 19287162 TI - Copy number variation on the human Y chromosome. AB - The Y chromosome is unusual in being constitutively haploid and escaping recombination for most of its length. This has led to a correspondingly unusual genomic landscape, rich in segmental duplications, which provide a potent environment for the generation of copy number variation (CNV). Interest in the chromosome comes from diverse fields, including infertility research, population genetics, forensics, and genealogy. Together with inclusion in more systematic surveys, this has led to the ascertainment of a variety of CNVs. Assessment in the context of the well-resolved Y phylogeny allows their mutational history to be deciphered and an estimation of mutation rate. The functional consequences of variants are moderated by the specialization of the chromosome and the presence of functionally equivalent X-chromosomal homologues for some genes. However, deletions of the AZFa, b, and c regions cause impaired spermatogenesis, while partial deletions and duplications within these regions, and deletions and duplications elsewhere, may be selectively neutral or have subtle phenotypes. PMID- 19287164 TI - Copy number variation and mosaicism. AB - Mosaicism is the presence of cells within an organism that have a different genetic composition despite deriving from a single zygote. The consequence of this depends on the number and type of cells that are affected as well as the specific DNA involved. There are several diseases where mosaicism is known to occur, but the currently observed frequency is presumably an underestimation due to the difficulty of detecting changes in only a percentage of cells. Recent advances in technology have provided a greater insight into the frequency and mechanisms of mosaicism in all forms. This review will cover the different techniques that can be used for detecting copy number variation (CNV) in mosaic form, and describe some of the insights that different diseases have given on the true frequencies and mechanisms responsible for somatic rearrangements. It will conclude with a discussion of the implications of the recent description of CNV in identical twins, and what remains to be elucidated in the world of mosaic CNV. PMID- 19287163 TI - Genomic drift and copy number variation of chemosensory receptor genes in humans and mice. AB - Recent studies about the structural variation of genomic sequences have shown that there is a large amount of copy number variations (CNVs) of genes within species. Analyzing Redon et al.'s (2006) crude data on copy number variable regions (CNVRs), we previously showed that CNVs are particularly high for chemosensory receptor genes in human populations. In this paper, we reanalyzed the CNVs of these genes using more refined data by Perry et al. (2008). The results showed that the extent of CNVs is somewhat lower in this dataset than in the previous one, but that the extent is still substantial for olfactory receptor (OR), vomeronasal receptor (VR), and taste receptor (TR) genes. We also studied the CNVs for chemosensory receptor genes in mice, using CNVR data obtained from inbred strains. It was found that the extent of CNVs is quite substantial but is lower than that for human populations. However, because the mouse data came from inbred strains and might be biased, this conclusion should be regarded as tentative. Despite this reservation, the distribution of gene copy number for the OR gene family was approximately normal in both humans and mice, suggesting that genomic drift caused by random duplication and deletion of genes plays important roles in determining the evolutionary change of chemosensation. PMID- 19287165 TI - Expression divergence and copy number variation in the human genome. AB - DNA copy number variation (CNV) was recently discovered as a significant part of human genetic variation. This variation affects genes as well as intergenic regions. Herein, current insight into the effects of CNV on gene expression is summarized. The consequences of intergenic CNVs are poorly understood. For CNV affecting genes, a dosage compensation mechanism seems to be applied on a subset of genes only, while others show augmented expression caused by increasing copy number. For the latter case, extensive literature reports a positive correlation between gene copy number and gene expression. Recent advances in techniques for genome-wide measurements of CNV and gene expression are described. These advances will soon allow the generation of comprehensive maps of these two phenomena, which will contribute to our understanding of the mechanisms underlying how CNV affects gene expression. PMID- 19287166 TI - The evolutionary significance of copy number variation in the human genome. AB - Copy number variation provides the raw material for gene family expansion and diversification, which is an important evolutionary force. Moreover, copy number variants (CNVs) can influence gene transcriptional and translational levels and have been associated with complex disease susceptibility. Therefore, natural selection may have affected at least some of the greater than one thousand CNVs thus far discovered among the genomes of phenotypically normal humans. While identifying and understanding particular instances of natural selection may shed light on important aspects of human evolutionary history, our ability to analyze CNVs in traditional population genetic frameworks has been limited. However, progress has been made by adapting some of these frameworks for use with copy number data. Moving forward, these efforts will be aided by non-human organism studies of the population genetics of copy number variation, and by more direct comparisons of within-species copy number variation and between-species copy number fixation. PMID- 19287167 TI - Comparative analysis of copy number variation in primate genomes. AB - Copy number differences between human and chimpanzee genomic sequences often overlap with regions of intraspecies copy number variation. Copy number variations (CNVs) that occur at orthologous sites in both humans and chimpanzees ('shared' CNVs) are likely to represent unstable genomic regions that have been prone to recurrent rearrangements during primate evolution. Lineage-specific CNVs are also important because they may have been subject to positive selection in specific primate lineages and hence could have contributed both to intra- and interspecies phenotypic diversity. In this study, we reinvestigated the data originally obtained by Perry et al. (2006) relating to chimpanzee CNVs and identified 24 genomic regions with the potential to be chimpanzee-specific CNVs. Since every putative chimpanzee-specific CNV was found in at least two of the 20 chimpanzees originally studied, it would appear as if these 24 CNVs are fairly frequent in chimpanzees. A combination of new mutation, genetic drift, and directional or balancing selection is likely to have influenced the maintenance of these CNVs in the chimpanzee population. Several genes map to the regions encompassed by the chimpanzee-specific CNVs. Some of their human orthologues are already known either to influence the phenotype (e.g., SLC24A4) or to be associated with inherited diseases (e.g., PAK3 and DTNBP1). Although the relatively small number of chimpanzee-specific CNVs (N = 24) among the 355 CNVs originally identified may be in part due to the use of a human BAC array to detect them, we nevertheless surmise that lineage-specific CNVs are not abundant in the chimpanzee. The thorough characterization of CNVs in the great ape genomes is a sine qua non for identifying the human-specific CNVs that may constitute genomic regions which have experienced either positive or negative selection during human evolution. PMID- 19287168 TI - Copy number variation in the mouse genome: implications for the mouse as a model organism for human disease. AB - Individuals within a species have genetic differences which ultimately result in the spectrum of phenotypic variation that we observe. Genetic variation exists at the nucleotide level in the form of single nucleotide polymorphisms (SNPs), and at a structural level as inversions, deletions and amplifications of larger stretches of nucleotides. Profiling of human and mouse genomes has identified numerous genomic segmental copy number variations (CNVs) throughout these genomes. Since inbred mice are widely used laboratory models for the study of both normal and disease biology, it is crucial that we understand the full scope of genetic variation, including CNVs, within these animals. These genetic differences can inform us about the history of a population or species, enlighten us on gene function, and guide our selection of a model system for the study of human disease. PMID- 19287169 TI - CNV discovery using SNP genotyping arrays. AB - Genome-wide single nucleotide polymorphism (SNP) genotyping platforms have made an important contribution to population genetics and genetic epidemiology. Recently there has been a realisation that these SNP platforms can also be used for typing copy number variants (CNVs). This allows for 'generalised' genotyping of both SNPs and CNVs simultaneously on a common sample set, with advantages in terms of cost and unified analysis. In this article we review various statistical approaches to calling CNVs from SNP data. We highlight three tiers of algorithms depending on the level of information used. PMID- 19287170 TI - Methods to detect CNVs in the human genome. AB - The detection of quantitative changes in genomic DNA, i.e. deletions and duplications or Copy Number Variants (CNVs), has recently gained considerable interest. First, detailed analysis of the human genome showed a surprising amount of CNVs, involving thousands of genes. Second, it was realised that the detection of CNVs as a cause of genetic disease was often neglected, but should be an essential part of a complete screening strategy. In both cases new efficient CNV screening methods, covering the entire range from specific loci to genome-wide, were behind these developments. This paper will briefly review the methods that are available to detect CNVs, discuss their strong and weak points, show some new developments and look ahead. Methods covered include microscopy, fluorescence in situ hybridization (including fiber-FISH), Southern blotting, PCR-based methods (including MLPA), array technology and massive parallel sequencing. In addition, we will show some new developments, including a 1400-plex CNV bead assay, fast MLPA (from DNA to result in approximately 6 h) and a simple Melting Curve Analysis assay to confirm potential CNVs. Using the 1400-plex CNV bead assay, targeting selected chromosomal regions only, we detected confirmed rearrangements in 9% of 320 mental retardation patients studied. PMID- 19287171 TI - Detection, breakpoint identification and detailed characterisation of a CNV at the FRA16D site using SNP assays. AB - Copy Number Variants (CNV) and other submicroscopic structural changes are now recognised to be widespread across the human genome. We show that SNP data generated for association study can be utilised for the identification of deletion CNVs. During analysis of data for an SNP association study for Specific Language Impairment (SLI) a deletion was identified. SLI adversely affects the language development of children in the absence of any obvious cause. Previous studies have found linkage to a region on chromosome 16. The deletion was located in a known fragile site FRA16D in intron 5-6 of the WWOX gene (also known as FOR). Changes in the FRA16D site have been previously linked to cancer and are often characterised in cell lines. A long-range PCR assay was used to confirm the existence of the deletion. We also show the breakpoint identification and large scale characterisation of this CNV in a normal human sample set. PMID- 19287172 TI - Methods to detect and analyze copy number variations at the genome-wide and locus specific levels. AB - Copy number variations (CNVs) have effects on phenotypes by altering transcription levels of genes and may have major impacts on protein sequence, structure and function. Therefore, CNV screening and analysis focused on the identification of CNV-genetic disease relations are actively progressing. CNVs can be detected and analyzed by various methodologies at the genome-wide and locus-specific levels. The genome-wide analysis of CNVs has been enhanced by bioinformatic tools for long-range sequence analysis, and comparative genome hybridization using microarrays containing either single nucleotide polymorphisms or bacterial artificial chromosome clones that represent the whole genome. RFLP followed by Southern blot analysis, quantitative real-time PCR, pyrosequencing, ligation detection reaction and the invader assay have become the main tools for locus-specific analysis so far. In this review, we present a brief principle, application history, and strengths and weaknesses of the methods used to detect CNVs at the genome-wide and locus-specific levels. PMID- 19287173 TI - Computational methods for identification of recurrent copy number alteration patterns by array CGH. AB - Recurrent DNA copy number alterations (CNA) are widely studied in diagnostic and cytogenetic cancer research. CNAs reveal locations that may alter gene dosage and thus expression of the genes contained within. Array comparative genomic hybridization has emerged as a popular high-throughput, genome-wide technique to interrogate tumor genomes for copy number alterations. When studying a group of tumors derived from a patient cohort, it is of great interest to detect the copy number alterations that are common across the population and thus assumed to be potential diagnostic markers and/or predictors of clinical outcome. In this paper, we review extant and available computational approaches for detecting such recurrent copy number alterations from array comparative genomic hybridization (aCGH) data. This is a challenging computational problem due to various sources of noise in the data that can obscure the recurrent copy number signals or induce false positives in their prediction. In this paper, we qualitatively evaluate methods designed to detect recurrent copy number alterations for aCGH data based on their analytical strengths and limitations, and discuss expected future directions in this important area of cancer research. PMID- 19287174 TI - Successful outcome after prenatal treatment of a cardiac diverticulum with massive pericardial effusion. AB - Congenital ventricular cardiac diverticula are rare. They may occur prenatally in association with a pericardial effusion which, if large enough, can compromise fetal circulatory and lung development. Parental counseling is difficult because some cases resolve in the second trimester and others progress to worsening hydrops and intrauterine death. We present a case associated with a massive pericardial effusion that had a good outcome after successful pericardiocentesis relatively late in gestation considering the time of critical pulmonary development. We also review similar cases presented in the literature and discuss management options. PMID- 19287175 TI - The effect of rate control on speech rate and intelligibility of dysarthric speech. AB - PURPOSE: This study investigated the effect of rate control methods (RCMs) on speaking rate (SR), articulation rate (AR), and intelligibility in dysarthric speakers. METHOD: Nineteen dysarthric patients (7 unilateral upper motor neuron dysarthria, 6 hypokinetic, 3 flaccid, 3 ataxic) participated. SR, AR and intelligibility ratings were determined on the basis of 1-min recorded reading passages. Seven RCMs were applied: voluntary rate control, hand tapping, alphabet board, pacing board and delayed auditory feedback with a delay of 50, 100 and 150 ms. RESULTS: Almost all methods resulted in lower mean SRs and ARs (p < 0.05). Rate control did not improve overall intelligibility of the dysarthric population. However, a meaningful increase of intelligibility was found in 5 participants. This study indicates that the effect of rate control on intelligibility is independent of habitual speech rate and type of dysarthria. Degree of intelligibility may be an influencing factor. The most effective methods are: voluntary rate control, alphabet board, hand tapping and pacing board. CONCLUSION: RCMs do result in lower speech rates. Some dysarthric individuals do benefit from one or more RCMs, but rate control may also have an inverse effect on intelligibility. PMID- 19287176 TI - Vocal symptoms in telemarketers: a random and controlled field trial. AB - The present study was carried out to evaluate the effectiveness of a specific program regarding the occurrence of vocal attrition symptoms in telemarketers. A total of 71 subjects participated in this study: 28 completed the Vocal Symptoms questionnaire to test its reliability, and 43 were randomly assigned to two groups: an 8-week vocal training group (n = 14) and a no-training control group (n = 29), to evaluate the effectiveness of the training program with this tool. The voice training group also filled in the posttraining questionnaire 'Benefits Obtained with Voice Training' (BVT). The vocal training program was not considered effective with regard to the occurrence of vocal symptoms. However, due to a probable increase in symptoms in untrained telemarketers, it can work as a protective factor. According to BVT answers, the vocal training contributed to an improvement in vocal use as a communication tool for telemarketers. PMID- 19287177 TI - Voice handicap and health-related quality of life in laryngectomees: assessments with the use of VHI and EORTC questionnaires. AB - AIMS: To investigate the Voice Handicap Index (VHI), the health-related quality of life (HRQL), and the correlations between VHI and HRQL in laryngectomees. METHODS: Forty-three laryngectomized persons participated (mean age 68;6 years, time since laryngectomy between 0;6 and 12 years). Evaluation of voice handicap was done with the VHI. HRQL was evaluated with questionnaires from the European Organization for Research and Treatment of Cancer, EORTC QLQ-C30 and EORTC QLQ H&N35. RESULTS: VHI for the whole group demonstrated a moderate voice handicap, with a mean score of 48/120. The functional scales of EORTC QLQ-C30 resulted in scores on the same level as the normal population with the exception of a lower global quality of life scale (Global QOL). EORTC QLQ-H&N35 revealed problems with smell and taste, speech, coughing, xerostomia, and sexuality. VHI correlated significantly with the Global QOL, the functional scales, dyspnea, pain, nausea and financial difficulties (EORTC QLQ-C30). Significant correlations were also found between VHI and speech problems, social contact, pain from the head and neck area, sense problems, sexuality and social eating (EORTC QLQ-H&N35). CONCLUSION: The EORTC questionnaires in combination with the VHI questionnaire seem to capture most of the problems following laryngectomy, including voice problems. PMID- 19287178 TI - Effect of second pregnancy on maternal carriage and outcome of high-risk human papillomavirus (HPV). Experience from the prospective finnish family HPV study. AB - OBJECTIVES: The effect of second pregnancy on human papillomavirus (HPV) carriage and outcome is modelled in longitudinal setting covering two subsequent pregnancies. STUDY DESIGN: Among 329 (baseline pregnant) women prospectively followed up in the Finnish Family HPV Study, two subcohorts were compiled: (i) 78 women (Group B) who became pregnant for the second time during the follow-up, and (ii) 100 women (Group A) who did not develop 2nd pregnancy. The effect of pregnancy on high-risk HPV (HR-HPV) carriage and outcome was analysed using Kaplan-Meier and Cox survival analyses and generalized estimating equation (GEE) modelling of the longitudinal data. RESULTS: Women in the two groups were similar in their baseline HR-HPV status but significantly different in several known risk factors of HR-HPV infection. Group A women showed higher point prevalence of cervical and oral HR-HPV at the 36-month (p = 0.015) and 6-month (p = 0.024) follow-up visit, respectively. Among Group B women, prevalence of both cervical and oral HR-HPV significantly decreased during 2nd pregnancy (p = 0.005 and p = 0.002) as compared with inter-pregnancy period, but increased again after 2nd pregnancy. There was no difference in acquisition or clearance of cervical or oral HR-HPV between the two groups in univariate (Kaplan-Meier) or multivariate (Cox) survival analysis. In the GEE approach, 2nd pregnancy was not significantly associated with cervical or oral HR-HPV carriage or persistence when adjusted for all other covariates. CONCLUSIONS: Second pregnancy is of little impact on carriage and persistence of oral and cervical HR-HPV infections in a longitudinal setting over time. PMID- 19287179 TI - Hemojuvelin: the hepcidin story continues. AB - Hemojuvelin (HJV) is a membrane protein that is responsible for the iron overload condition known as juvenile hemochromatosis. HJV, highly expressed in the liver, skeletal muscle and heart, seems to play a role in iron absorption and release from cells and has anti-inflammatory properties. HJV is a bone morphogenetic protein (BMP) co-receptor and signals via the SMAD (human homolog of Drosophila mad--mother against decapentaplegic) pathway to regulate hepcidin expression. HJV acts as a BMP co-receptor. Moreover, HJV plays an essential role in the regulation of hepcidin expression, specifically in the iron-sensing pathway, although through unknown mechanisms. Dietary iron sensing and inflammatory pathways converge in the regulation of the key regulator hepcidin, but how these two pathways intersect remains unclear. Inflammation, through downregulation of hepatic HJV, might induce temporary elimination of iron sensing. Despite enormous scientific achievements in explaining the pathogenetic mechanisms of iron metabolism, many questions still remain unanswered: What is the functional role of HJV in iron metabolism? How it is related to hepcidin expression in different settings? How do iron-sensing and inflammatory pathways cooperate in hepcidin gene expression? PMID- 19287180 TI - Serum neutrophil gelatinase-associated lipocalin as a marker of renal function in patients with chronic heart failure and coronary artery disease. AB - Heart failure and chronic kidney disease share a number of risk factors and pathophysiological pathways. Renal insufficiency is common in patients with chronic heart failure (CHF). The aim of the study was to assess whether neutrophil gelatinase-associated lipocalin (NGAL) could represent a novel, sensitive marker of kidney function in adult patients with chronic heart failure and normal serum creatinine. The study was performed on 150 patients with chronic heart failure due to coronary artery disease. Serum and urinary NGAL as well as serum cystatin C were measured using commercially available kits. Serum NGAL was related, in univariate analysis, to serum creatinine, urinary NGAL, hemoglobin, hematocrit, leukocyte count, eGFR, cystatin C. Urinary NGAL correlated with age, hemoglobin, hematocrit, serum creatinine, eGFR. In multiple regression analysis predictors of serum NGAL were NYHA class, cystatin C, and eGFR. Taking into consideration the fact that the recent DOQI states that individuals with a reduced GFR is at greater risk for cardiovascular disease and cardiac deaths, precise evaluation of renal function is important in order to select the appropriate strategy to reduce the cardiovascular risk. NGAL should be investigated as a potential early and sensitive marker of kidney impairment/injury. PMID- 19287181 TI - Tripterygium wilfordii Hook F treatment for idiopathic refractory nephrotic syndrome in adults: a meta-analysis. AB - Tripterygium wilfordii Hook F (TwHF) is a Chinese herbal medicine with immunosuppressive effects and an established history of use in the treatment of inflammatory and autoimmune diseases. We have carried out a meta-analysis of randomized controlled trials (RCTs) and quasi-RCTs which assessed the effectiveness of TwHF in the treatment for idiopathic refractory nephrotic syndrome (IRNS). After reading the full text, only three RCTs or quasi-RCTs meeting our inclusion criteria were selected. Our meta-analysis indicated that TwHF has beneficial effects on the remission of IRNS. There was no significant publication bias in the meta-analysis studies. However, studies with a larger sample size and including patients with both a mild and severe histopathological change involvement separately are needed to assess the effects of TwHF in IRNS patients before exact conclusions can be drawn. PMID- 19287182 TI - Can pulsatile cardiopulmonary bypass prevent perioperative renal dysfunction during myocardial revascularization in elderly patients? AB - BACKGROUNDS/AIMS: We recently demonstrated that pulsatile cardiopulmonary bypass (CPB) versus standard linear CPB is associated with better perioperative renal function. Since older subjects have a higher risk of acute renal failure, we have extended our study to evaluate the specific impact of pulsatile CPB on the perioperative renal function in elderly patients. METHODS: We enrolled 50 patients with normal preoperative renal function: they were stratified by age (65 75 vs. 50-64 years) and randomized to nonpulsatile (group A) or pulsatile CPB (group B). Twenty-seven patients aged > or =50 years and <65 years were randomized to group A (n = 12) or to group B (n = 15) and 23, aged > or =65 years and < or =75 years, to group A (n = 13) or to group B (n = 10). Glomerular filtrate rate (GFR), daily diuresis, lactatemia and other parameters were measured during the pre- and perioperative period. RESULTS: The percent perioperative decrease in GFR was lower in group A than in group B (p < 0.001), without differences between older and younger patients. By contrast, perioperative plasma lactate levels were higher in group A than in group B (p < 0.001), both in older and younger patients. No difference was observed for 24 h urine output and blood urea nitrogen. CONCLUSIONS: Pulsatile CPB preserves renal function better than standard CPB even in patients older than 65. CPB could be adopted as the procedure of choice in this subgroup of patients. PMID- 19287183 TI - Statins do not affect mineral metabolism in chronic kidney disease: a retrospective analysis. AB - It has recently been proposed that statins act as vitamin D analogs in binding the ubiquitously expressed vitamin D receptor, accounting for the perceived pleiotropic effects of statins (a reduction in cancer risk, prevention of organ transplant rejection and autoimmune disease). Chronic kidney disease (CKD) offers a useful test of this hypothesis: serum 25-hydroxyvitamin D levels are insufficient (<75 nmol/l) in as many as 76% of patients with advanced CKD, associated with secondary hyperparathyroidism and reduced bone mineralization. Vitamin D suppresses parathyroid hormone (PTH) secretion in part through its action on the vitamin D receptor. If statins act as vitamin D analogs, they may then be able to suppress PTH secretion in CKD. We examined data on 714 vitamin D analog naive patients with stage 3-4 CKD. 404 patients were treated with a statin indicated almost exclusively for primary prevention of coronary heart disease, and 310 patients were not. Both groups were similar in characteristics. Statins had no effect on the intact PTH concentration, the percentage of patients achieving K/DOQI PTH targets, or on calcium or phosphate concentrations. In patients with stage 3-4 CKD, statins had no effect on secondary hyperparathyroidism. If the hypothesis contending that statins act as vitamin D analogs to exert pleiotropic effects is true, this is of no clinical benefit in the prevention of secondary hyperparathyroidism in CKD. PMID- 19287184 TI - Recent racial/ethnic disparities in stroke hospitalizations and outcomes for young adults in Florida, 2001-2006. AB - BACKGROUND: Black-white disparities in stroke mortality are well documented, but few recent studies have examined racial/ethnic disparities in stroke hospitalizations among young adults. We analyzed recent (2001-2006) trends in stroke hospitalizations and hospital case-fatality for black, Hispanic, and white adults aged 25-49 years in Florida. METHODS: Hospitalization rates were calculated using population estimates from the census, and hospital discharges with a primary diagnosis of stroke (ICD-9-CM 430, 431, 434, 436) (n = 16,317). Multivariate logistic regression modeling was used to examine racial/ethnic disparities in stroke mortality prior to discharge, after adjustment for patient sociodemographics, stroke subtype, risk factors, and comorbidities. RESULTS: Age adjusted stroke hospitalization rates for blacks were over 3 times higher than rates for whites, while rates for Hispanics were slightly higher than rates for whites. Hemorrhagic strokes were proportionally greater among Hispanics compared with blacks and whites (p < 0.0001). Blacks were most likely to have diagnosed hypertension (62.3%), morbid obesity (10.9%) or drug abuse (13.6%). Whites were most likely to have diagnosed hyperlipidemia (21.0%), alcohol abuse (9.5%), and to be smokers (30.6%). The in-hospital fatality rate for all strokes was highest among blacks (10.0%) compared with whites (9.0%) and Hispanics (8.2%). After adjustment for age, gender, insurance status, and all diagnosed risk factors and comorbidities, the black excess was no longer observed [odds ratio (OR) 1.01, 95% confidence interval (CI) 0.88-1.15, p = 0.93]. However, the Hispanic advantage in case-fatality was strengthened (OR 0.66, 95% CI 0.55-0.79, p < 0.0001). Separate case-fatality analyses for ischemic versus hemorrhagic strokes yielded similar results. CONCLUSIONS: Our study found a strong and persistent black-white disparity in stroke hospitalization rates for young adults. In contrast, rates were similar for Hispanics and whites. Multivariate adjustment explained the 15% excess case-fatality for blacks; the short-term mortality advantage among Hispanics was strengthened after adjustment. PMID- 19287185 TI - Racial/ethnic differences in stroke in young adults. PMID- 19287186 TI - Improving vaccines by targeting antigens to dendritic cells. AB - A new approach to enhancing the effectiveness of vaccines is to deliver antigens selectively to dendritic cells (DC) in situ, via monoclonal antibodies specific for particular DC surface molecules. This can markedly enhance CTL responses and, via helper T cells, also enhance antibody responses. DC activation agents or adjuvants must also be administered for effective CTL responses, but in some cases good antibody responses can be obtained without adjuvants. Here we review the role of different DC subsets and different DC target molecules in obtaining enhanced immune responses. PMID- 19287187 TI - Myocardial infarction patients show altered lipoprotein properties and functions when compared with stable angina pectoris patients. AB - Several parameters and risk factors were compared between Korean male myocardial infarction (MI) patients (n=10) and angina pectoris (AP) patients (n=17) to search unique biomarkers for myocardial infarction (MI) in lipoprotein level. Individual serum and lipoprotein fractions (VLDL, LDL, HDL2, HDL3) were isolated and analyzed by lipid and protein determination and enzyme assay. The MI group was found to have a 25 and 30% higher serum cholesterol and triacylglycerol (TG) level than the AP group, respectively, however, their body mass index (BMI), LDL cholesterol (C), HDL-C, and glucose levels fell within the normal range. MI patients were found to have an approximately two-fold higher level of serum IL-6 and an 18% lower serum apoA-I level than that of the AP group. LDL and HDL2 fraction of the MI group were more enriched with TG than those of AP group. The increased TG was correlated well with the increased level of apoC-III in the same fraction. Cholesteryl ester transfer protein (CETP) activity and protein level were greatly increased in MI patients in the LDL and HDL3 fractions. MI patients showed more severely oxidized LDL fraction than patients in the AP group, as well as the weakest antioxidant ability of serum. Conclusively, MI patients were found to have unique serum and lipoprotein characteristics including increased IL-6 and TG in serum, with CETP and apoC-III in the LDL and HDL fractions, as well as severely impaired antioxidant ability of HDL. PMID- 19287188 TI - Expression analyses of human cleft palate tissue suggest a role for osteopontin and immune related factors in palatal development. AB - Cleft lip and/or palate (CL/P) is a common congenital malformation with a complex etiology which is not fully elucidated yet. Epidemiological studies point to different etiologies in the cleft lip and palate subgroups, isolated cleft lip (CL), isolated cleft palate (CP) and combined cleft lip and palate (CLP). In order to understand the biological basis in these cleft lip and palate subgroups better we studied the expression profiles in human tissue from patients with CL/P. In each of the CL/P subgroups, samples were obtained from three patients and gene expression analysis was performed. Moreover, selected differentially expressed genes were analyzed by quantitative RT-PCR, and by immunohistochemical staining of craniofacial tissue from human embryos. Osteopontin (SPP1) and other immune related genes were significantly higher expressed in palate tissue from patients with CLP compared to CP and immunostaining in palatal shelves against SPP1, chemokine receptor 4 (CXCR4) and serglycin (PRG1) in human embryonic craniofacial tissue were positive, supporting a role for these genes in palatal development. However, gene expression profiles are subject to variations during growth and therefore we recommend that future gene expression in CL/P studies should use tissue from the correct embryonic time and place if possible, to overcome the biases in the presented study. PMID- 19287189 TI - Extracellular HIV-1 Tat up-regulates expression of matrix metalloproteinase-9 via a MAPK-NF-kappaB dependent pathway in human astrocytes. AB - The infiltration of monocytes into the CNS represents one of the early steps to inflammatory events in AIDS-related encephalitis and dementia. Increased activity of selected matrix metalloproteinases (MMPs) such as MMP-9 impairs the integrity of blood-brain barrier leading to enhanced monocyte infiltration into the CNS. In this study, we examined the effect of HIV-1 Tat on the expression of MMP-9 in CRT MG human astroglioma cells. Treatment of CRT-MG cells with HIV-1 Tat protein significantly increased protein levels of MMP-9, as measured by Western blot analysis, zymography and an ELISA. Treatment of CRT-MG cells with HIV-1 Tat protein markedly increased mRNA levels of MMP-9, as analyzed by RT-PCR. Pretreatment of CRT-MG cells with NF-kappaB inhibitors led to decrease in Tat induced protein and mRNA expression of MMP-9. Pretreatment of CRT-MG cells with MAPK inhibitors suppressed Tat-induced MMP-9 expression. Furthermore, HIV-1 Tat induced expression of MMP-9 was significantly inhibited by neutralization of TNF alpha, but not IL-1beta and IL-6. Taken together, our results indicate that HIV-1 Tat can up-regulate expression of MMP-9 via MAPK-NF-kappaB-dependent mechanisms as well as Tat-induced TNF-alpha production in astrocytes. PMID- 19287190 TI - STAT3 is involved in phosphatidic acid-induced Bcl-2 expression in HeLa cells. AB - Phosphatidic acid (PA), the product of a PLD-mediated reaction, is a lipid second messenger that participates in various intracellular signaling events and is known to regulate a growing list of signaling proteins. We found that Bcl-2 was upregulated by PA treatment in HeLa cells. However, how PA upregulates Bcl-2 expression has not yet been studied. In this study, we tried to discover the mechanisms of Bcl-2 up-regulation by PA treatment in HeLa cells. Treatment with PA resulted in significantly increased expression of Bcl-2 in HeLa cells. Moreover, PA-induced Bcl-2 expression was blocked by mepacrine, an inhibitor of PLA2, but not by propranolol, an inhibitor of PA phospholyhydrolase (PAP). Treatment of 1,2-dipalmitoryl-sn-glycero-3- phosphate (DPPA) also increased Bcl-2 expression. These results indicate that Bcl-2 expression is mediated by lysophosphatidic acid (LPA), not by arachidonic acid (AA). Thereafter, we used MEK1/2 inhibitor, PD98059 to investigate the relationship between ERK1/2 MAPK and PA-induced Bcl-2 expression. PA-induced Bcl-2 expression was decreased when ERK1/2 was inhibited by PD98059. The transcription factor such as STAT3 which is controlled by ERK1/2 MAPK was increased along with Bcl-2 expression when the cells were treated with PA. Furthermore, STAT3 siRNA treatments inhibited PA induced Bcl-2 expression, suggesting that STAT3 (Ser727) is involved in PA induced Bcl-2 expression. Taken together, these findings indicate that PA acts as an important mediator for increasing Bcl-2 expression through STAT3 (Ser727) activation via the ERK1/2 MAPK pathway. PMID- 19287191 TI - Genetic and expression alterations in association with the sarcomatous change of cholangiocarcinoma cells. AB - Cholangiocarcinoma (CC) is an intrahepatic bile duct carcinoma with a high mortality rate and a poor prognosis. Sarcomatous change/epithelial mesenchymal transition (EMT) of CC frequently leads to aggressive intrahepatic spread and metastasis. The aim of this study was to identify the genetic alterations and gene expression pattern that might be associated with the sarcomatous change in CC. Previously, we established 4 human CC cell lines (SCK, JCK1, Cho-CK, and Choi CK). In the present study, we characterized a typical sarcomatoid phenotype of SCK, and classified the other cell lines according to tumor cell differentiation (a poorly differentiated JCK, a moderately differentiated Cho-CK, and a well differentiated Choi-CK cells), both morphologically and immunocytologically. We further analyzed the genetic alterations of two tumor suppressor genes (p53 and FHIT) and the expression of Fas/FasL gene, well known CC-related receptor and its ligand, in these four CC cell lines. The deletion mutation of p53 was found in the sarcomatoid SCK cells. These cells expressed much less Fas/FasL mRNAs than did the other ordinary CC cells. We further characterize the gene expression pattern that is involved in the sarcomatous progression of CC, using cDNA microarrays that contained 18,688 genes. Comparison of the expression patterns between the sarcomatoid SCK cells and the differentiated Choi-CK cells enabled us to identify 260 genes and 247 genes that were significantly over-expressed and under-expressed, respectively. Northern blotting of the 14 randomly selected genes verified the microarray data, including the differential expressions of the LGALS1, TGFBI, CES1, LDHB, UCHL1, ASPH, VDAC1, VIL2, CCND2, S100P, CALB1, MAL2, GPX1, and ANXA8 mRNAs. Immunohistochemistry also revealed in part the differential expressions of these gene proteins. These results suggest that those genetic and gene expression alterations may be relevant to the sarcomatous change/EMT in CC cells. PMID- 19287192 TI - Bone morphogenetic protein 4 stimulates neuronal differentiation of neuronal stem cells through the ERK pathway. AB - Bone morphogenic protein 4 (BMP4), a member of the TGF-beta superfamily, induced neural differentiation of neural stem cells (NSCs) grown in a medium containing basic fibroblast growth factor (bFGF). The Ras protein level and the activities of the downstream ERKs were increased by transfection of BMP4 or treatment with recombinant BMP4. The effects of BMP4, including activation of the Ras-ERK pathway and induction of the neuron marker beta-tubulin type III (Tuj1), were blocked by co-treatment of the BMP4 antagonist, noggin. The roles of the Ras-ERK pathway in neuronal differentiation by BMP4 were revealed by measuring the effect of the ERK pathway inhibition by dominant negative Ras or PD98059, the MEK specific inhibitor. BMP4 is a transcriptional target of Wnt/beta-catenin signaling, and both the mRNA and protein levels of BMP4 were increased by treatment of valproic acid (VPA), a chemical inhibitor of glycogen synthase kinase 3beta (GSK3beta) activating the Wnt/beta-catenin pathway. The BMP4- mimicking effects of VPA, activation of the Ras-ERK pathway and induction of Tuj1, also were blocked by noggin. These results indicate the potential therapeutic usage of VPA as a replacement for BMP4. PMID- 19287193 TI - High glucose, unsaturated and saturated fatty acids differentially regulate expression of ATP-binding cassette transporters ABCA1 and ABCG1 in human macrophages. AB - The ATP-binding cassette transporters ABCA1 and ABCG1 are highly expressed in macrophage-derived foam cells and promote reverse cholesterol efflux via biogenesis of high-density lipoproteins. The aim of this study was to analyze the direct effects of bioactive factors related to the metabolic syndrome on macrophage transcript levels of all 47 human ABC transporters. Using in vitro M CSF predifferentiated macrophages and TaqMan low density arrays we could show that linoleic acid, palmitic acid, and high glucose levels have a major impact on ABCA1 and ABCG1 expression but do not strongly affect most other human ABC transporters. In Western blot experiments we demonstrate that ABCA1 and ABCG1 protein levels are synchronously suppressed by high glucose levels and the w6 unsaturated fatty acid linoleic acid. We conclude that metabolites associated with the metabolic syndrome enhance the formation of atherosclerotic lesions by diminishing the reverse cholesterol transport function of ABCA1 and ABCG1. PMID- 19287194 TI - Effects of a chemical chaperone on genetic mutations in alpha-galactosidase A in Korean patients with Fabry disease. AB - Fabry disease is an X-linked inborn error of glycosphingolipid catabolism that results from mutations in the gene encoding the alpha-galactosidase A (GLA) enzyme. We have identified 15 distinct mutations in the GLA gene in 13 unrelated patients with classic Fabry disease and 2 unrelated patients with atypical Fabry disease. Two of the identified mutations were novel (i.e., the D231G missense mutation and the L268delfsX1 deletion mutation). This study evaluated the effects of the chemical chaperones 1-deoxygalactonojirimycin (DGJ) on the function of GLA in vitro, in cells containing missense mutations in the GLA gene. Nine missense and a nonsense mutations, including one novel mutation were cloned into mammalian expression vectors. After transient expression in COS-7 cells, GLA enzyme activity and protein expression were analyzed using fluorescence spectrophotometry and Western blot analysis, respectively. DGJ enhanced GLA enzyme activity in the M42V, I91T, R112C and F113L mutants. Interestingly, the I91T and F113L mutations are associated with the atypical form of Fabry disease. However, DGJ treatment did not have any significant effect on the GLA enzyme activity and protein expression of other mutants, including C142W, D231G, D266N, and S297F. Of note, GLA enzyme activity was not detected in the novel mutant (i.e., D231G), although protein expression was similar to the wild type. In the absence of DGJ, the E66Q mutant had wild-type levels of GLA protein expression and approximately 40% GLA activity, indicating that E66Q is either a mild mutation or a functional single nucleotide polymorphism (SNP). Thus, the results of this study suggest that the chemical chaperone DGJ enhances GLA enzyme activity and protein expression in milder mutations associated with the atypical form of Fabry disease. PMID- 19287195 TI - Functional characterization of human oncoprotein gankyrin in Zebrafish. AB - Gankyrin is an oncoprotein containing seven ankyrin repeats that is overexpressed in hepatocellular carcinoma (HCC). Gankyrin binds to Mdm2, which results in accelerated ubiquitylation via degradation of p53, and it also plays an important role in cell proliferation. However, little is known about the relationships between p53 levels, cell proliferation, and gankyrin over-expression. In order to investigate the influence of gankyrin protein on p53 and Mdm2 in a zebrafish model, we injected human gankyrin (hgankyrin) containing expression vectors (pCS2 hgankyrin, pCS2- hgankyrin-EGFP) into zebrafish embryos. To measure p53 and Mdm2 expression in hgankyrin-injected embryos, RT-PCR, Northern blot and in-situ hybridization and BrdU immunostaining were used. In addition, to know the effect of hgankyrin on cell proliferation in vitro, cell viability assays such as MTT, trypan blue staining and RT-PCR following transfection of hgankyrin-containing vector into HEK 293 cell line were performed. In vivo results indicated that p53 mRNA levels decreased but those of Mdm2 were not decreased in the presence of hgankyrin. These results suggest that gankyrin downregulates p53 expression and not Mdm2 expression. In the study of cell proliferation, BrdU-positive cells were predominantly increased in the head and tail regions in hgankyrin-injected zebrafish. Additional in vitro studies using trypan blue staining and MTT assay showed that gankyrin-expressing HEK 293 cells proliferated at a faster rate, indicating that gankyrin promotes cell proliferation. Our results demonstrate that hgankyrin overexpression downregulates p53 expression and promotes cell proliferation in zebrafish. Gankyrin may play an important role in tumorigenesis via its effects on p53 and cell proliferation. PMID- 19287196 TI - Thromboxane A(2) modulates migration, proliferation, and differentiation of adipose tissue-derived mesenchymal stem cells. AB - Prostanoid metabolites are key mediators in inflammatory responses, and accumulating evidence suggests that mesenchymal stem cells (MSCs) can be recruited to injured or inflamed tissues. In the present study, we investigated whether prostanoid metabolites can regulate migration, proliferation, and differentiation potentials of MSCs. We demonstrated herein that the stable thromboxane A(2) (Tx(2)) mimetic U46619 strongly stimulated migration and proliferation of human adipose tissue-derived MSCs (hADSCs). Furthermore, U46619 treatment increased expression of alpha-smooth muscle actin (alpha-SMA), a smooth muscle marker, in hADSCs, suggesting differentiation of hADSCs into smooth muscle like cells. U46619 activated ERK and p38 MAPK, and pretreatment of the cells with the MEK inhibitor U0126 or the p38 MAPK inhibitor SB202190 abrogated the U46619 induced migration, proliferation, and alpha-SMA expression. These results suggest that TxA2 plays a key role in the migration, proliferation, and differentiation of hADSCs into smooth muscle-like cells through signaling mechanisms involving ERK and p38 MAPK. PMID- 19287197 TI - Fabrication of an alpha-lipoic acid-eluting poly-(D,L-lactide-co-caprolactone) cuff for the inhibition of neointimal formation. AB - The purpose of this study was to develop a novel polymer cuff for the local delivery of alpha-lipoic acid (ALA) to inhibit neointimal formation in vivo. The polymer cuff was fabricated by incorporating the ALA into poly- (D,L-lactide-co caprolactone) 40:60 (PLC), with or without methoxy polyethylene glycol (MethoxyPEG). The release kinetics of ALA and in vitro degradation by hydrolysis were analyzed by HPLC and field emission scanning electron microscopy (FE-SEM), respectively. In vivo evaluation of the effect of the ALA-containing polymer cuff was carried out using a rat femoral artery cuff injury model. At 24 h, 48% or 87% of the ALA was released from PCL cuffs with or without MethoxyPEG. FE-SEM results indicated that ALA was blended homogenously in the PLC with MethoxyPEG, whereas ALA was distributed on the surface of the PLC cuff without MethoxyPEG. The PLC cuff with MethoxyPEG showed prolonged and controlled release of ALA in PBS, in contrast to the PLC cuff without MethoxyPEG. Both ALA-containing polymer cuffs had a significant effect on the inhibition of neointimal formation in rat femoral artery. Novel ALA-containing polymer cuffs made of PLC were found to be biocompatible and effective in inhibiting neointimal formation in vivo. Polymer cuffs containing MethoxyPEG allowed the release of ALA for one additional week, and the rate of drug release from the PLC could be controlled by changing the composition of the polymer. These findings demonstrate that polymer cuffs may be an easy tool for the evaluation of anti-restenotic agents in animal models. PMID- 19287198 TI - Association of TBX21 polymorphisms in a Korean population with rheumatoid arthritis. AB - TBX21 (T-bet) is a member of the T-box family of transcriptional factors that contain a conserved DNA binding domain. TBX21 is a critical regulator of the commitment to the Th1 lineage and IFN-gamma production. Th1 and Th2 cells cross regulate the differentiation of each other, and in this way TBX21 could be an attractive candidate gene for treating autoimmune disease such as rheumatoid arthritis (RA). In present study, we analyzed the genotypic frequencies of six polymorphisms of the TBX21 gene between the 367 RA patients and the 572 healthy controls. We showed that the g.-1514T>C and c.99C>G polymorphisms are suggestively associated with RA susceptibility. It is interesting that the genotypic frequencies of the TBX21 polymorphisms (g.-1514T>C and c.2103A>C) in the male RA patients were significantly different from the male control group (P=0.0016 and 0.045, respectively). We also found that the g.-1514T>C and c.2103A>C polymorphisms of the TBX21 gene in the male RA patients have significant association with the levels of anti-CCP (P=0.05) and rheumatoid factor (P=0.03), respectively. These results suggest that the polymorphisms of the TBX21 gene might be associated with the susceptibility to male RA patients. PMID- 19287199 TI - Expression of N-terminal truncated desmoglein 3 (deltaNDg3) in epidermis and its role in keratinocyte differentiation. AB - During a search for keratinocyte differentiation-related genes, we obtained a cDNA fragment from the 5'-untranslated region of a previously identified splicing variant of desmoglein 3 (Dg3). This transcript encodes a protein of 282 amino acids, which corresponds to the N-terminal truncated intracellular domain of Dg3 (deltaNDg3). Northern blot analysis detected a 4.6-kb transcript matching the predicted size of deltaNDg3 mRNA, and Western blot analysis with an antibody raised against the Dg3 C-terminus (H-145) detected a 31-kDa protein. Increased deltaNDg3 expression was observed in differentiating keratinocytes by RT-PCR and Western blot analysis, suggesting that deltaNDg3 is indeed a differentiation related gene product. In immunohistochemical studies of normal and pathologic tissues, H-145 antibody detected the protein in the cytoplasm of suprabasal layer cells, whereas an antibody directed against the N-terminal region of Dg3 (AF1720) reacted with a membrane protein in the basal layer. In addition, deltaNDg3 transcript and protein were upregulated in psoriatic epidermis, and protein expression appeared to increase in epidermal tumors including Bowen's disease and squamous cell carcinoma. Moreover, overexpression of deltaNDg3 led to increased migration and weakening of cell adhesion. These results suggest that deltaNDg3 have a role in keratinocyte differentiation, and that may be related with tumorigenesis of epithelial origin. PMID- 19287200 TI - Regulation of glucose metabolism-related genes and VEGF by HIF-1alpha and HIF 1beta, but not HIF-2alpha, in gastric cancer. AB - Hypoxia-inducible factors (HIFs) are transcription factors that activate the transcription of target genes involved in crucial aspects of cancer development. This study investigated the expression of HIFs and their contribution to the regulation of target genes related to angiogenesis and glucose metabolism in gastric cancer. The data showed that HIFs were over-expressed in gastric cancer and that activation of the target genes was observed mainly in the early stages. Moreover, the results of the present study revealed that only HIF-1alpha, but not HIF-2alpha dimerizes with HIF-1beta and then regulates expression of target genes in response to hypoxia. The results of the present study demonstrate that HIF 1alpha and HIF-1beta enhances expression of VEGF and glucose metabolism-related genes in response to hypoxia in gastric cancer. These data offer important information regarding HIF pathways in the development of gastric cancer. PMID- 19287201 TI - Transcription factor sequestration by nuclear envelope components. PMID- 19287202 TI - Understanding protective immune mechanisms induced by malaria vaccines in the context of clinical trials. AB - Malaria is one of the world's most important diseases, and an effective, licensed vaccine would be a major public health tool. There is currently a surge in efforts to develop malaria vaccines, and many candidates are entering and progressing through clinical trials. However, inadequate understanding of host immunity, gaps in knowledge of parasite biology and the lack of surrogate markers of protection have hindered rational vaccine development and led to failure to control malaria. A greater understanding of naturally acquired immunity would certainly help guide vaccine development and deployment. The research tools at our disposal are greater and more superior than before, and the growing capacity to perform immunological assays in the field brings new possibilities for investigating vaccine-induced immunity and immune correlates of protection within clinical trials. The malaria vaccine community needs to learn from the TB and HIV vaccine communities, who have developed and optimized multi-parameter immunological assays, some of which are amenable for studies conducted in African children. The best strategy is to take advantage of large clinical trials in endemic countries to better understand immunity to malaria, vaccine take and identify surrogates of protection. PMID- 19287203 TI - Rac1 regulates cardiovascular development and postnatal function of endothelium. AB - Rac1 is a member of the small Rho GTPase family, which controls actin cytoskeleton and focal adhesion dynamics in cellular protrusions. While Rac1 therefore contributes to regulation of endothelial cell-cell and cell-matrix interactions, a detailed understanding of its role in endothelium function is lacking. Recently, the role of Rac1 in development and postnatal regulation of the cardiovascular system has been investigated in murine models lacking Rac1 specifically in endothelium. Homozygous endothelial deletion was lethal, primarily due to defects in angiogenesis. Rac1-deficient endothelial cells were unable to form cellular protrusions/lamellipodia, leading to impaired cell-cell and cell-matrix interactions, and resulting in dysfunctional adhesion, motility, permeability and capillary morphogenesis. Development was normal in the heterozygous model, however a hypertensive phenotype was observed as a result of reduced nitric oxide signalling. Nitric oxide synthase activity was regulated by Rac1 at multiple levels; expression, mRNA stability and uptake of the nitric oxide synthase substrate L-arginine. Therefore, Rac1 activity is essential in regulating developmental and postnatal angiogenesis and cardiovascular function, by controlling nitric oxide production, and formation of endothelial cell protrusions. PMID- 19287204 TI - SRF in angiogenesis: branching the vascular system. AB - Cell cytoskeleton proteins are fundamental to cell shape, cell adhesion and cell motility, and therefore play an important role during angiogenesis. One of the major regulators of cytoskeletal protein expression is serum response factor (SRF), a MADS-box transcription factor that regulates multiple genes implicated in cell growth, migration, cytoskeletal organization, energy metabolism and myogenesis. Recent data have demonstrated a crucial role of SRF downstream of VEGF and FGF signalling during sprouting angiogenesis, regulating endothelial cell (EC) migration, actin polymerisation, tip cell morphology, EC junction assembly and vascular integrity. Here, we review the role of SRF in the regulation of angiogenesis and EC function, integrate SRF function into a broader mechanism regulating branching morphogenesis, and discuss future directions and perspectives of SRF in EC biology. PMID- 19287205 TI - The role of the transcriptional regulator snail in cell detachment, reattachment and migration. AB - In order to metastasize, cancer cells must first detach from the primary tumor, migrate, invade through tissues and attach to a second site. The transcription factor snail is an important mediator of epithelial-mesenchymal transitions and is involved in tumor progression. Recent data have provided evidence for a requirement for snail expression in metastatic dissemination. Although very little is known about the molecular mechanisms governing metastatic dissemination, we review the possible roles of snail expression in this process. We also review the regulation of snail expression. PMID- 19287207 TI - The eukaryotic translation initiation factor 4E (eIF4E) and HuR RNA operons collaboratively regulate the expression of survival and proliferative genes. PMID- 19287206 TI - Functional diversity of SDF-1 splicing variants. AB - SDF-1 is ubiquitously expressed in vertebrate tissues in a constitutive manner. It performs an essential role in cell migration and proliferation as well as participates in tissue-specific physiological processes such as neuromodulation. It is also involved in many pathological processes including: HIV infection, metastatic malignancy, chronic inflammatory disorders and benign proliferative diseases. SDF-1 is mostly regulated at the splicing, and not transcriptional level. Different splicing variants share agonist potency to their cognate receptor, CXCR4, but are characterized by distinct properties. SDF-1alpha is the predominant isoform found in all organs, but undergoes rapid proteolysis in blood. SDF-1beta is more resistant to blood-dependent degradation, stimulates angiogenesis and is present in highly vascularized organs such as: the liver, spleen and kidneys. In contrast, SDF-1gamma is located in very active, less vascularized organs susceptible to infarction such as the heart and the brain. The understanding of the functional diversity of the different splicing variants will help in developing therapeutic strategies. PMID- 19287208 TI - TORCs/CRTCs: more than mere coincidence. PMID- 19287209 TI - Enhancing radiosensitivity: targeting the DNA repair pathways. AB - Radiotherapy is very effective in local control of cancerous tumors, but its curative potential is often limited by intrinsic radioresistance of the tumor cells. Since DNA repair pathways remove radiation-induced DNA lesions and protect cells from lethality, these pathways represent potential therapeutic targets to radiosensitize tumors. In order to achieve a therapeutic gain, however, there must be a differential between tumor and normal cells that can be exploited to preferentially target the DNA repair of the tumor, while sparing surrounding normal tissues, and this has represented a significant challenge to progress. Nevertheless, recent advances in our understanding of DNA repair mechanisms and tumor biology, on both the biochemical and genetic levels, have identified molecular differentials that may increase tumor specificity. This mechanistic insight suggests new strategies for radiotherapeutic targeting of DNA repair. Some of these strategies are reviewed here, including synthetic lethal, replicative stress, cell cycle and hypoxia-based approaches. The example of PARP1 inhibitor use in BRCA1 and 2 mutated breast cancer therapy is discussed, and future directions and challenges are explored. PMID- 19287210 TI - Monocyte attachment to laminin in diabetes mellitus: The role of ATP. AB - Monocyte-extracellular matrix interactions have been implicated in atherosclerosis pathophysiology. Laminin, the main basement membrane protein contains cell binding domains that can be cryptic, presented only after protein modification. In the present study we evaluated monocyte attachment to laminin-1 in the presence of ATP. Monocytes were derived from either healthy volunteers or patients with diabetes mellitus type II. For the estimation of monocyte attachment to laminin the myeloperoxidase assay was used. Monocytes derived from diabetic patients, showed an increased ability to attach to laminin (p = 0.0055). The presence of ATP increased the attachment of control monocytes to laminin (p = 0.0022). On the contrary, the presence of ATP did not affect the attachment of monocytes derived from diabetic patients to laminin. Our results indicate a modified interaction between monocytes and laminin-1 in diabetes mellitus. PMID- 19287211 TI - A novel, human Atg13 binding protein, Atg101, interacts with ULK1 and is essential for macroautophagy. AB - Macroautophagy is an intracellular, vesicle-mediated mechanism for the sequestration and ultimate lysosomal degradation of cytoplasmic proteins, organelles and macromolecules. The macroautophagy process and many of the autophagy-specific (Atg) proteins are remarkably well conserved in higher eukaryotes. In yeast, the Atg1 kinase complex includes Atg1, Atg13, Atg17, and at least four other interacting proteins, some of which are phosphorylated in a TOR dependent manner, placing the Atg1 signaling complex downstream of a major nutrient-sensing pathway. Atg1 orthologs, including mammalian unc-51-like kinase 1 (ULK1), have been identified in higher eukaryotes and have been functionally linked to autophagy. This suggests that other components of the Atg1 complex exist in higher eukaryotes. Recently, a putative human Atg13 ortholog, FLJ20698, was identified by gapped-BLAST analysis. We show here that FLJ20698 (Atg13) is a ULK1-interacting phosphoprotein that is essential for macroautophagy. Furthermore, we identify a novel, human Atg13-interacting protein, FLJ11773, which we have termed Atg101. Atg101 is essential for autophagy and interacts with ULK1 in an Atg13-dependent manner. Additionally, we present evidence that intracellular localization of the ULK1 complex is regulated by nutrient conditions. Finally, we demonstrate that Atg101 stabilizes the expression of Atg13 in the cell, suggesting that Atg101 contributes to Atg13 function by protecting Atg13 from proteasomal degradation. Therefore, the identification of the novel protein, Atg101, and the validation of Atg13 and Atg101 as ULK1 interacting proteins, suggests an Atg1 complex is involved in the induction of macroautophagy in mammalian cells. PMID- 19287212 TI - Epigenetic mechanisms in sequential differentiation of neural stem cells. AB - The major cell types in the central nervous system, neurons and glial cells (astrocytes and oligodendrocytes), are generated from common neural stem cells (NSCs). However, neurogenesis precedes gliogenesis, which normally commences at later stages of fetal brain development. Recent studies have shown that epigenetic gene regulation, in concert with cues from outside the cells, plays important roles in regulating the differentiation potential and fate specification of NSCs. In this review, we discuss emerging molecular explanations for the transitions that occur during the stepwise process of NSC fate specification. PMID- 19287213 TI - Multi-drug resistance protein 4 (MRP4/ABCC4) and cyclic nucleotides signaling pathways. PMID- 19287214 TI - Hemimethylation footprints of DNA demethylation in cancer. AB - Hypomethylation of DNA repeats, including satellite 2 DNA (Sat2), is one of the most frequent epigenetic changes in cancer. We examined ovarian epithelial tumors and diverse control tissues for methylation on only one strand (hemimethylation), both strands (symmetrical methylation), or neither strand at Sat2 CpG dyads using hairpin genomic sequencing. Analysis of the resulting cloned DNA molecules indicated that although carcinomas displayed much symmetrical hypomethylation of CpG dyads, there was cancer-linked hypermethylation at one of the thirteen dyads in the examined 0.2 kb Sat2 region. Hemimethylated sites were seen in both carcinomas and controls but, importantly, in carcinoma DNA molecules, they were significantly more likely to occur in clusters displaying the same orientation (the same strand methylated). Our data suggest that hemimethylated CpG dyads are intermediates in active demethylation during carcinogenesis and not just due to a failure of maintenance methylation during replicative DNA synthesis. Constitutive heterochromatin may be especially suitable for providing a snapshot of demethylation intermediates because hemimethylation might be more long-lived in heterochromatin due to its highly condensed state. PMID- 19287215 TI - Integrin switching modulates adhesion dynamics and cell migration. AB - When cells are stimulated to move, for instance during development, wound healing or angiogenesis, they undergo changes in the turnover of their cell-matrix adhesions. This is often accompanied by alterations in the expression profile of integrins-the extracellular matrix receptors that mediate anchorage within these adhesions. Here, we discuss how a shift in expression between two different types of integrins that bind fibronectin can have dramatic consequences for cell-matrix adhesion dynamics and cell motility. PMID- 19287216 TI - Specific antitumor effects of tumor vaccine produced by autologous dendritic cells transfected with allogeneic osteosarcoma total RNA through electroporation in rats. AB - OBJECTIVE: Transfection of dendritic cells with tumor-derived RNA has recently been shown to elicit tumor-specific CTL capable of recognizing and lysing a variety of tumor cells. However, the integration of allogeneic osteosarcoma mRNA and autologous DCs has not been fully examined. This study was designed to investigate the antitumor effects of tumor vaccine produced by autologous dendritic cells transfected with allogeneic osteosarcoma total RNA through electroporation in tumor-bearing rats model. RESULTS: The immunization using autologous DCs electrotransfected with allogeneic osteosarcoma total RNA induced UMR106-specific CTL responses which were statistically significant and the cytotoxic activity was inhibited by the treatment with anti-CD8 and anti-MHC class I monoclonal antibodies. In in vivo experiments, 80% of the rats immunized with allogeneic osteosarcoma RNA transfected to DCs were typically able to reject tumor challenge and remained tumor-free. Vaccinated survivors developed long immunological memory and were able to reject a subsequent rechallenge with the same tumor cells but not a syngeneic unrelated tumor line. METHODS: In the present study, we transfected Wistar rat osteosarcoma cells derived total RNA to SD rat bone marrow-derived DCs through electroporation. The tumor vaccine was applied to in tumor-bearing rats model and the specific antitumor effects of the tumor vaccine were observed. Then CTL activity was evaluated one week after the first immunization of SD rats with electroporated DCs and the specificity of the cytotoxic activity was confirmed in cold target inhibition assays and using mAbs blocking MHC class I or CD8 molecules. CONCLUSION: The present study provided valid evidence of integration of rat allogeneic tumor-derived mRNA and autologous DCs through electroporation and confirmed this novel tumor vaccine have the potential to induced osteosarcoma-specific CTL response and reject osteosarcoma challege. This technique and its products may thus represent a promising strategy for DC-based immunotherapy of patients with osteosarcoma. PMID- 19287242 TI - Verification of consumers' experiences and perceptions of genetic discrimination and its impact on utilization of genetic testing. AB - PURPOSE: To undertake a systematic process of verification of consumer accounts of alleged genetic discrimination. METHODS: Verification of incidents reported in life insurance and other contexts that met the criteria of genetic discrimination, and the impact of fear of such treatment, was determined, with consent, through interview, document analysis and where appropriate, direct contact with the third party involved. The process comprised obtaining evidence that the alleged incident was accurately reported and determining whether the decision or action seemed to be justifiable and/or ethical. RESULTS: Reported incidents of genetic discrimination were verified in life insurance access, underwriting and coercion (9), applications for worker's compensation (1) and early release from prison (1) and in two cases of fear of discrimination impacting on access to genetic testing. Relevant conditions were inherited cancer susceptibility (8), Huntington disease (3), hereditary hemochromatosis (1), and polycystic kidney disease (1). In two cases, the reversal of an adverse underwriting decision to standard rate after intervention with insurers by genetics health professionals was verified. The mismatch between consumer and third party accounts in three life insurance incidents involved miscommunication or lack of information provision by financial advisers. CONCLUSION: These first cases of verified genetic discrimination make it essential for policies and guidelines to be developed and implemented to ensure appropriate use of genetic test results in insurance underwriting, to promote education and training in the financial industry, and to provide support for consumers and health professionals undertaking challenges of adverse decisions. PMID- 19287244 TI - SQUIRE Guidelines for reporting improvement studies in healthcare: implications for nursing publications. PMID- 19287243 TI - Clinical outcomes after long-term treatment with alglucosidase alfa in infants and children with advanced Pompe disease. AB - PURPOSE: A clinical trial was conducted to evaluate the safety and efficacy of alglucosidase alfa in infants and children with advanced Pompe disease. METHODS: Open-label, multicenter study of IV alglucosidase alfa treatment in 21 infants 3 43 months old (median 13 months) with minimal acid alpha-glucosidase activity and abnormal left ventricular mass index by echocardiography. Patients received IV alglucosidase alfa every 2 weeks for up to 168 weeks (median 120 weeks). Survival results were compared with an untreated reference cohort. RESULTS: At study end, 71% (15/21) of patients were alive and 44% (7/16) of invasive-ventilator free patients remained so. Compared with the untreated reference cohort, alglucosidase alfa reduced the risk of death by 79% (P < 0.001) and the risk of invasive ventilation by 58% (P = 0.02). Left ventricular mass index improved or remained normal in all patients evaluated beyond 12 weeks; 62% (13/21) achieved new motor milestones. Five patients were walking independently at the end of the study and 86% (18/21) gained functional independence skills. Overall, 52% (11/21) of patients experienced infusion-associated reactions; 95% (19/20) developed IgG antibodies to recombinant human lysosomal acid alpha-glucosidase; no patients withdrew from the study because of safety concerns. CONCLUSIONS: In this population of infants with advanced disease, biweekly infusions with alglucosidase alfa prolonged survival and invasive ventilation-free survival. Treatment also improved indices of cardiomyopathy, motor skills, and functional independence. PMID- 19287245 TI - Importance of recommendations for gestational weight gain. PMID- 19287246 TI - Experience with implementation of a quality improvement project for the care of nursing home residents. PMID- 19287247 TI - The Magnet Model as a framework for excellence. PMID- 19287248 TI - Ticket to ride: reducing handoff risk during hospital patient transport. AB - A significant number of medical errors occur during patient handoffs, leading to less than optimal care, patient harm, and even death. The Joint Commission National Patient Safety Goals require hospitals to implement a standardized approach to "handoff" communications. The authors describe an initiative that focuses on standardization of the physical and informational handoff during hospital transport; outcomes are promising in terms of both patient safety and patient satisfaction. PMID- 19287249 TI - Why do chronic venous leg ulcers not heal? AB - Chronic venous leg ulcers account for the majority of lower extremity ulcers and represent a serious health and financial burden. Although several clinical practice guidelines and systematic reviews are available to guide treatment decisions, healing rates have not improved over the last decade. This research study illustrates some of the limitations of relying on existing databases to guide wound management as well as the importance of considering patient preferences, staff knowledge and skill, and available resources. PMID- 19287251 TI - Body mass index, weight, and pressure ulcer prevalence: an analysis of the 2006 2007 International Pressure Ulcer Prevalence Surveys. AB - To assess the relationships among pressure ulcer prevalence, body mass index (BMI), and weight, this report analyzed the US data from the 2006 and 2007 International Pressure Ulcer Prevalence Surveys. Findings indicated an overall reduction in pressure ulcer prevalence from 2004 and 2005 to 2006 and 2007; there was a higher prevalence of pressure ulcers in patients with low BMI and patients with both low and high weights. One in 10 patients were extremely obese. PMID- 19287252 TI - Bedside handover: quality improvement strategy to "transform care at the bedside". AB - This quality improvement project implemented bedside handover in nursing. Using Lewin's 3-Step Model for Change, 3 wards in an Australian hospital changed from verbal reporting in an isolated room to bedside handover. Practice guidelines and a competency standard were developed. The change was received positively by both staff and patients. Staff members reported that bedside handover improved safety, efficiency, teamwork, and the level of support from senior staff members. PMID- 19287253 TI - Standardization of change-of-shift report. AB - This article describes a clinical nurse-led initiative that changed the traditional group shift report in the conference room to a combination of a written report with a nurse-to-nurse verbal exchange at the patient's bedside. The new process resulted in less time spent in shift report, financial savings from reduced overtime, and a decrease in the number of patient falls and call lights during change of shift. PMID- 19287254 TI - Nurse satisfaction and creation of an admission, discharge, and teaching nurse position. AB - The goal of this study was to evaluate the effect of a newly developed nurse admission, discharge, and teaching position on nurse satisfaction and the quality of the admission and discharge process. A pretest-posttest design was used to collect data on nurse satisfaction, workload, and medication reconciliation. Study results supported the use of an admission, discharge, and teaching nurse to improve nurse satisfaction and the quality of the admission and discharge process. PMID- 19287255 TI - Evaluation of quality improvement initiative in pediatric oncology: implementation of aggressive hydration protocol. AB - Our goal was to improve the efficiency of chemotherapy administration for pediatric oncology patients. We identified prechemotherapy hydration as the process that most often delayed chemotherapy administration. An aggressive hydration protocol, supported by fluid order sets, was developed for patients receiving planned chemotherapy. The mean interval from admission to achieving adequate hydration status was reduced significantly from 4.9 to 1.4 hours with a minor reduction in the time to initiate chemotherapy from 9.6 to 8.6 hours. Chemotherapy availability became the new rate-limiting process. PMID- 19287256 TI - A quality improvement project to optimize patient outcomes following the maze procedure. AB - Atrial fibrillation (AF) is the most common of all clinically sustained heart arrhythmias with associated morbidities (shortness of breath, fatigue, and stroke). The maze cardiac surgical procedure is a new treatment option available for patients who have medical refractory AF. The purpose of this study was to determine whether a written postdischarge protocol was necessary to improve outcomes following the maze procedure. Only 3 (27%) patients with AF were actively treated by an arrhythmia protocol for restoration of sinus rhythm. Unnecessary pharmacologic management for AF was found in 44 patients with normal sinus rhythm. A postdischarge protocol was developed that improved outcomes. PMID- 19287257 TI - Leveraging safer nursing care by conceptualizing near misses as recovery processes. AB - The purpose of this study was to elicit perceptions and experiences of nurses associated with a recovery process (near miss) intervention (wound care management). From the content analysis of interview data involving 12 registered nurses, 2 key themes emerged. Both themes illuminated the links among pattern recognition, alignment of care processes and practice guidelines, and nurses' accountability for safe pressure ulcer care. PMID- 19287258 TI - Facilitating patients' diabetes self-management: a primary care intervention framework. AB - A quality primary care framework for diabetes self-management (DSM) is presented on the basis of prevalent office setting changes. Recent DSM research and literature are reviewed concerning (a) patients' DSM perceptions and office visit experiences and (b) more effective office-based DSM programs. This quality primary care framework enables nurse clinicians to more consistently assess and integrate individual patient factors using typically limited clinic resources into office visits to optimize DSM patient outcomes. PMID- 19287259 TI - Context for WOC practice: synthesizing the evidence to guide facility wide policies for wound, ostomy and continence care. PMID- 19287261 TI - Data collection methods series: part 2: select the most feasible data collection mode. PMID- 19287262 TI - Nursing interventions to reduce the risk of catheter-associated urinary tract infection: part 2: staff education, monitoring, and care techniques. AB - BACKGROUND: The US Centers for Medicare & Medicaid Services has enacted 2 policies that have focused considerable attention on the optimal use of indwelling catheters in the acute and long-term care settings and the prevention of complications including catheter-associated urinary tract infection (CAUTI). OBJECTIVES: This is the second of a 2-part Evidence-Based Report Card reviewing current evidence pertaining to nursing actions for prevention of CAUTI in patients with short- and long-term indwelling catheters. Part 2 reviews multiple interventions for CAUTI prevention including staff education, monitoring of catheter use and CAUTI incidence, insertion technique, urethral meatal care, securement, use of a closed drainage system, bladder irrigation, frequency of catheter change, and antiseptic solutions in the drainage bag. SEARCH STRATEGY: Nursing actions for prevention of CAUTI were identified based on search of electronic databases and Web-based search engines for national or international clinical practice guidelines focusing on this topic. Evidence related to the above nursing interventions was identified by searching electronic databases MEDLINE, CINAHL, the Cochrane Library, the ancestry of articles identified in these searches and Google scholar. RESULTS: Limited evidence suggests that the following interventions reduce the incidence of CAUTI in patients managed by short-term indwelling catheterization: (1) staff education about catheter management, combined with regular monitoring of CAUTI incidence, (2) a facility wide program to ensure catheterization only when indicated and prompt removal of indwelling catheters, (3) daily cleansing of the urethral meatus using soap and water or perineal cleanser, and (4) maintenance of a closed urinary drainage system. Mixed evidence suggests that use of a preconnected system reduces inadvertent interruption of a closed urinary drainage system and may prevent CAUTI. Limited evidence suggests that routine catheter changes every 4 to 6 weeks reduce CAUTI incidence in patients managed by long-term catheterization. Existed evidence suggests that the following interventions are not effective for reducing CAUTI incidence: (1) use of sterile technique for catheter insertion, (2) use of antiseptic solutions or ointments during routine meatal care, (3) use of a 2 chambered urinary drainage bag, (4) use of antiseptic filters incorporated into a urinary drainage bag, (5) bladder or catheter irrigation, (6) frequent changes of the urinary drainage bag, and (7) placement of an antiseptic solution in the urinary drainage bag. IMPLICATIONS FOR PRACTICE: Evidence from parts 1 and 2 of this Evidence-Based Report Card provides a sound basis for designing an evidence based program to prevent CAUTI. Essential elements of a CAUTI prevention program include staff education, ongoing monitoring of CAUTI incidence, monitoring catheter insertion and ensuring prompt removal, and careful attention to techniques for catheterization and catheter care. PMID- 19287263 TI - Catheter-associated urinary tract infections: fact sheet. PMID- 19287264 TI - Reducing the risks of wound consultation: adding digital images to verbal reports. AB - PURPOSE: The purpose of this study was to examine the impact of digital images on the assessment and recommendations of a WOC nurse who was providing remote nurse to-nurse consultations on home care patients with wounds. METHODS: In a descriptive comparative study, data were collected by home care nurses from a sample of 43 adult patients with a total of 89 wounds with various etiologies. To determine whether or not the addition of a digital photograph influenced the WOC nurse's assessment and recommendations, the WOC nurse first completed a wound assessment and recommendation form based on a verbal report from the home care nurse. The WOC nurse then accessed digital images of the wounds and made any indicated modifications to the original assessment and management plan, providing a rationale for any changes. Comparisons were made between the assessment completed by the home care nurse and the WOC nurse's assessment and between the WOC nurse's assessment and recommendations based only on a verbal report and his or her assessment and recommendations based on the combination of a verbal report and a digital photograph. RESULTS: Although there was a high percentage of agreement between the wound assessments completed by the home care nurse and those completed by the WOC nurse, areas of disagreement often impacted on the overall assessment. The agreement rates between the WOC nurse's assessment and recommendations based only on a verbal report versus those based on a combination of verbal report and digital photographs were as follows: total agreement (26/89 = 29.2%), trivial disagreement (11/89 = 12.4%), and clinically relevant disagreement (52/89 = 58.4%). CONCLUSIONS: WOC nurses who provide remote nurse-to nurse consultations without directly visualizing the patients' wounds through digital images are at risk for under-or overtreating patients' wounds. Digital images also provide an opportunity for the WOC nurse to mentor home care nurses in wound assessment and care. PMID- 19287265 TI - Adjuvant use of acoustic pressure wound therapy for treatment of chronic wounds: a retrospective analysis. AB - PURPOSE: Small studies have indicated that the addition of acoustic pressure wound therapy (APWT) to conventional wound care may hasten healing of chronic wounds. We evaluated our early clinical experience using APWT as an adjunct to conventional wound care. DESIGN: The study was a retrospective chart review of consecutive patients receiving APWT in addition to conventional wound care in a hospital-based, primarily outpatient setting. METHODS: Medical records of all patients treated with APWT between August 2006 and October 2007 were reviewed. Analysis included the 41 patients with 52 wounds who received APWT at least 2 times per week during the study period. Statistical comparisons were made for wound dimensions, tissue characteristics, and pain at start versus end of APWT. RESULTS: Thirty-eight percent of wounds (N = 20) healed completely with a mean 6.8 weeks of APWT. Median wound area and volume decreased significantly (88% [P < .0001] and 100% [P < .0001], respectively) from start to end of APWT. The proportion of wounds with greater than 75% granulation tissue increased from 26% (n = 12) to 80% (n = 41) (P < .0001), and normal periwound skin increased from 25% (n = 13) to 54% (n = 28) (P = .0001). Presence of greater than 50% fibrin slough decreased from 50% (n = 24) to 9% (n = 4) of wounds (P = .006). CONCLUSIONS: This early experience supplementing conventional wound care with APWT suggests it may promote healing in chronic wounds, where the ordered cellular and molecular processes leading to healing have stalled. PMID- 19287266 TI - Pressure ulcer awareness and prevention program: a quality improvement program through the Canadian Association of Wound Care. AB - The Canadian Association of Wound Care funded a study to determine the extent of pressure ulcers in Canada and discovered that the mean prevalence rate was 26%. Recognizing this as a significant health-related problem, the Canadian Association of Wound Care created a continuous quality improvement program known as the Pressure Ulcer Awareness and Prevention program to support a culture shift in healthcare settings, that is, shifting the emphasis on management from treatment of existing ulcers to their prevention. This program has been pilot tested, revised, and is being implemented in many Canadian healthcare facilities. The program is described and results are presented that demonstrate a reduction rate up to 57% (prevalence) and 71% (incidence). PMID- 19287267 TI - The improvement of wound-associated pain and healing trajectory with a comprehensive foot and leg ulcer care model. AB - PURPOSE: Pain is a major concern for subjects with chronic wounds, but its optimal management remains elusive. The aim of this study was to validate an organized pain management approach using the Wound Associated Pain model in subjects with chronic leg and foot ulcers. DESIGN: We completed a prospective cohort study that documented pain in chronic wound subjects over a 4-week period. SUBJECTS AND SETTING: A total of 111 subjects with chronic leg and foot ulcers were recruited from the community and ambulatory wound care clinics. RESULTS: Using a systematic approach based on the Wound Associated Pain model, we demonstrated improved overall wound healing outcomes in 111 subjects with chronic leg and foot ulcers. Using an 11-point numerical rating scale, the average level of pain was reduced from 6.3 at week 0 to 2.8 at week 4 (P < .001). The average healing rate was 0.39 cm per week and the average relative reduction in size was 59.36% (t = 2.31; P = .023). To examine the relationship between pain and wound healing, pain levels were compared in subjects who achieved wound closure and those who did not. The mean pain score was 1.67 for the healed subjects in contrast to 3.21 for those who did not achieve complete wound closure (P < .041). CONCLUSIONS: A comprehensive patient assessment can improve chronic leg and foot ulcer wound-related pain and healing rates. The mean pain scores are lower for patients with healed ulcers than for those who do not obtain complete wound closure. PMID- 19287269 TI - Knowledge and attitudes about urinary incontinence among community-dwelling Korean American women. AB - PURPOSE: The purpose of this study was to explore knowledge and attitudes about urinary incontinence (UI) among Korean American women with incontinence and provide initial information needed to design education programs and culture specific interventions. SUBJECTS AND SETTING: One hundred eighty-two community dwelling Korean American women who were 30 years and older and self-identified as having UI were invited to participate in the study. Data collection was conducted in 12 Korean religious organizations. DESIGN: This study uses a cross-sectional descriptive design by means of interviews. INSTRUMENTS: The Incontinence Quiz was used to measure knowledge and attitudes about UI. Higher scores indicate greater knowledge and more positive attitudes. RESULTS: The mean Incontinence Quiz was 4.85 (SD = 2.75) out of 14, which was much lower than the midpoint of 7.0, indicating that respondents tended to have limited knowledge and negative attitudes toward UI. The number of correct responses to the items on the Incontinence Quiz in this sample was lower than that reported in other studies that sampled the general population. CONCLUSIONS: Interventions to improve Korean women's knowledge of, and attitudes toward, UI are needed. WOC and continence nurses should take an active role in educating women about the prevention and treatment of UI. PMID- 19287270 TI - Ostomy: general principles of management and patient teaching. PMID- 19287271 TI - Options in negative pressure wound therapy: five case studies. PMID- 19287273 TI - Clinical, electrocardiographic, and laboratory findings in children with amitriptyline intoxication. AB - BACKGROUND: Amitriptyline is one of the major tricyclic antidepressants, and the data on amitriptyline poisoning in children are limited. OBJECTIVES: To present our experiences with amitriptyline poisoning in children, particularly with regard to its effects on electrocardiogram (ECG) and relation with clinical status. METHODS: Clinical, laboratory, and electrocardiographic findings in 52 children admitted with amitriptyline poisoning were reviewed. Patients were divided into 2 groups according to age, as 6 years or younger (group A) and older than 6 years (group B). RESULTS: Mean age was 4.6 +/- 3.0 years. Thirty-one patients were male, and 41 were 6 years or younger. Ingested amitriptyline dose was known in 23 patients (range, 2.3 mg/kg-27 mg/kg). The most frequent findings were lethargy (76.9%), sinus tachycardia (57.7%), and coma (48.1%). Four patients had a history of convulsion. The most common laboratory abnormalities were hyponatremia (26.9%) and leukocytosis (25%). Elevated transaminase levels were observed in 4 patients. In ECG, 11 (22.4%) patients had QTc prolongation and in 4 (8.2%) of them, it was significant. In 4 patients (8.2%), the QRS duration was 100 ms or longer and in 15 patients, the R wave in aVR was 3 mm or longer. The frequencies of clinical, laboratory, and electrocardiographic findings were similar between the 2 age groups (P > 0.05). No clinically apparent arrhythmias were observed. The positive predictive value of a widened QRS was 100% in terms of coma. None of the patients with an R wave in aVR of less than 3 mm developed convulsion; thus, the negative predictive value of an R wave in aVR of 3 mm or longer was 100% in terms of convulsion. CONCLUSIONS: Amitriptyline poisoning may result in severe toxicity. Frequencies of clinical, laboratory, and ECG findings were similar in the 2 age groups. Amitriptyline overdose results in some ECG changes that can possibly help to predict the results of poisoning. Absence of an R wave in aVR of 3 mm or longer predicts seizures with a high negative predictive value, and a QRS duration of 100 or longer ms predicts coma with a high positive predictive value. PMID- 19287274 TI - Cerebral sinovenous thrombosis masquerading as migraine: a case report. AB - Headaches are common during childhood and a common pediatric presentation to emergency departments (EDs). The most common diagnoses for acute headache in the pediatric ED are viral upper respiratory tract infection with fever, sinusitis, and migraine, accounting for 70% to 75% of presentations; serious neurologic diagnoses are rare. Most literature recommends against neuroimaging for uncomplicated headache given the absence of any other signs or symptoms associated with causes of intracranial pathology and the presence of a normal physical examination. Cerebral sinovenous thrombosis (CSVT) is a rare entity whose diagnosis relies heavily on neuroimaging and which carries high morbidity and mortality if untreated. Pediatric CSVT has yet to be presented in the emergency medicine literature. We report the case of a child who presented to the pediatric ED with headache, which was complicated by background difficulties with aggression and other behavioral issues. This case illustrates an extremely rare ED presentation of CSVT masquerading as migraine headache and discusses the diagnostic dilemma of neuroimaging. PMID- 19287275 TI - Diagnosis of pediatric intussusception by an emergency physician-performed bedside ultrasound: a case report. AB - The past decade has seen a rapid evolution in the use of bedside ultrasound (BUS) in the emergency department (ED). In addition, it has been increasingly used in the pediatric population for a variety of indications. As ED BUS is a relatively new modality in pediatric emergency medicine, novel indications are increasingly being recognized. We report a case of a 9-month-old infant who presented with signs and symptoms suggestive of intussusception, whose diagnosis was made using ED BUS in the pediatric ED. Knowledge of the sonographic appearance of intussusception can aid the emergency physician in the prompt diagnosis of this condition while potentially minimizing complications that may result from a delay in diagnosis. PMID- 19287276 TI - Retropharyngeal abscess with mediastinal extension in an infant--still existing? AB - OBJECTIVES: The objective of the present case report was to note that despite the low incidence of retropharyngeal abscess in the present antibiotic era, children, especially infants, can develop this condition. If early diagnosis is missed, the abscess may extend into the mediastinum and can be life threatening. METHODS: Case reports of mediastinal abscess in the past literature along with the current case. RESULTS: The present case was successfully managed by surgical drainage of the abscess via transcervical route along with intravenous antibiotic therapy, and the recovery was uneventful. CONCLUSIONS: Although the incidence of retropharyngeal abscess has declined with the widespread use of antibiotics, the presenting symptom can be overlooked in a pediatric population, especially infants, and can extend along the fascial planes to the mediastinum when the recovery becomes difficult. The present report is an effort to draw attention to this life-threatening complication of retropharyngeal abscess, emphasize the role of early diagnosis, and prompt surgical management. PMID- 19287277 TI - Acute appendicitis after a blunt perineal trauma: an illustrative case. AB - Acute appendicitis after abdominal trauma, although rare, has been described in the past. We herein report, for the first time, a case of acute appendicitis after a blunt trauma to the perineum in a boy. Clinicians taking care of children with injuries to the abdomen or perineal area should be aware of the possibility that they can develop posttraumatic acute appendicitis. PMID- 19287278 TI - A transatlantic caterpillar. AB - BACKGROUND: Saddleback caterpillar sting has been very rarely reported in European countries. We report a French case of a stung toddler. CASE: A 2-year old girl was brought to the pediatric emergency department after being stung by a caterpillar in a furniture store. The emergency department physical examination revealed an inflammatory edema of the stung finger, normal vital signs, and no fever. Three hours after presentation, she was discharged with local ointment prescription. The caterpillar brought in by the parents was unusual compared to common French caterpillar species. The regional poison center was unable to identify it. With the help of the Internet, we succeeded in its identification as a saddleback caterpillar. Tracing its transatlantic importation was the most difficult. CONCLUSIONS: Saddleback caterpillars can be imported to France and carried across the Atlantic Ocean on house or garden plants especially Areca trees. French garden store owners should be informed about this risk and should check and treat host plants (especially Areca trees) at the arrival time. PMID- 19287279 TI - Right atrial rupture from blunt thoracic trauma in a 4-year-old child. AB - Blunt cardiac injury in the pediatric population has been less frequently reported than in the adult population. Cardiac chamber rupture is a rare but highly lethal injury in both populations. The lethality of this condition is further enhanced by the multiplicity of injuries that are frequently present. The diagnosis of cardiac chamber rupture is often delayed while other injuries are being addressed. A high index of suspicion and early use of the appropriate imaging studies are essential for the timely diagnosis of this condition. In this report, we present a 4-year-old child who survived 10 hours of pericardial tamponade secondary to blunt right atrial rupture. PMID- 19287280 TI - Meningococcal group A sepsis associated with rare manifestations and complicated by Kawasaki-like disease. AB - Neisseria meningitidis serogroup A septicemia and the development of Kawasaki disease in a 10-month-old male infant are described. The patient also experienced a number of primary/septic manifestations as consequences of the direct infectious assault and secondary/hypersensitivity immune-mediated complications. The present observation indicates an etiological relationship between invasive meningococcal infection group A and Kawasaki disease, an association in favor of superantigen-induced theory for the latter occurrence. PMID- 19287281 TI - Straight from the spring? PMID- 19287282 TI - The risks and the identification of ingested button batteries in the esophagus: a child safety issue. PMID- 19287283 TI - Newer agents for rapid sequence intubation: etomidate and rocuronium. AB - The emergency airway management of children and adolescents with critical illnesses may necessitate rapid sequence intubation with a sedating and a neuromuscular blocking agent. Etomidate and rocuronium have become increasingly popular for the sedation and paralysis, respectively, of pediatric patients in rapid sequence intubation, and there are many advantages to the use of both agents. Both etomidate and rocuronium have a rapid onset of action, and both agents are relatively free of hemodynamic adverse effects. Etomidate does, however, suppress adrenal function, and consequently, its use in patients with septic shock is controversial. Rocuronium can produce optimal intubating conditions without the serious complications that can accompany succinylcholine. The available evidence supports the safety of etomidate and rocuronium in rapid sequence intubation but also suggests that more prospective studies are needed in pediatric patients. PMID- 19287284 TI - ECGs in the ED. PMID- 19287285 TI - Torticollis as an initial sign of Chiari I malformation. PMID- 19287286 TI - Efficacy of 1.25 MG versus 2.5 MG intravitreal bevacizumab for diabetic macular edema: six-month results of a randomized controlled trial. AB - PURPOSE: To evaluate the efficacy of intravitreal injections of two different dosages of bevacizumab (Avastin) for treating diffuse diabetic macular edema. METHODS: Fifty-two eyes of 52 patients with diabetic macular edema were randomized to receive three monthly intravitreal injections of 1.25 mg or 2.5 mg bevacizumab. Patients were observed for 6 months and optical coherence tomography central foveal thickness, logMAR best-corrected visual acuity (BCVA), and adverse events were assessed. RESULTS: Forty-eight eyes of 48 patients completed the 6 month follow-up and were analyzed. Significant mean central foveal thickness reductions were observed in both groups at all follow-up visits (P < 0.013). Significant improvements between baseline and 6-month mean logMAR BCVAs were seen, with the mean logMAR BCVA improved from 0.63 to 0.52 in the 1.25 mg group and 0.60 to 0.47 in the 2.5 mg group. No significant difference in BCVA was observed between the two groups at any time point (P > 0.56). Subgroup analysis showed that intravitreal bevacizumab seemed to be more effective in eyes without any previous diabetic macular edema treatment. CONCLUSIONS: Three monthly intravitreal bevacizumab injections resulted in significant reduction in central foveal thickness and improvements in BCVA in diabetic macular edema patients. Both 1.25 mg and 2.5 mg seemed to have similar treatment efficacy. PMID- 19287287 TI - Safety of repeat intravitreal injections of bevacizumab versus ranibizumab: our experience after 2,000 injections. AB - PURPOSE: To compare the safety of repeat intravitreal injections of bevacizumab versus ranibizumab performed on a large series of patients during the past 2 years period of time. METHODS: Four hundred fifty patients receiving 2,000 injections (1,275 bevacizumab and 725 ranibizumab) were studied retrospectively. Injections performed in a usual examination room under the standard sterile conditions. Follow-up varied from 3 to 24 months. RESULTS: Serious ocular adverse events were uncommon. Only one patient developed retinal detachment (0.05%). Most common procedure-related ocular adverse event was injection-site redness (64.75%). Postoperative subconjuctival hemorrhage occurred after 200 (10%) injections. Patients receiving aspirin treatment were more prone to have subconjuctival hemorrhage (P = 0.0002). Most common drug-related ocular adverse event was uveitis (1.90%), which was treated successfully and lasted no >12 days. There was no statistically significant difference between the patients treated with bevacizumab or ranibizumab regarding the noted adverse events (P > 0.5%). CONCLUSION: Multiple intravitreal injections of bevacizumab or ranibizumab were both well tolerated and safe. Performing injections on a usual examination room proved safe. Injection-site redness, subconjuctival hemorrhage, and uveitis were the most common ocular adverse events. Aspirin treatment was a risk factor for the development of subconjuctival hemorrhage. PMID- 19287288 TI - Intravitreal injection of 2.5 mg versus 1.25 mg bevacizumab (Avastin) for treatment of CNV associated with AMD. AB - PURPOSE: To compare the safety and efficacy of intravitreal injections of 1.25 and 2.5 mg bevacizumab for treatment of choroidal neovascularization associated with age-related macular degeneration. METHODS: In this prospective, randomized, comparative clinical trial, 86 patients with active choroidal neovascularization associated with age-related macular degeneration were studied. Baseline best corrected visual acuity in the study eye was from 20/40 to 20/2000. Patients were randomly assigned to receive intravitreal injections of 2.5 (39 patients) or 1.25 mg (47 patients) of intravitreal bevacizumab. Best-corrected visual acuity measurement and clinical ocular examination were performed at 1 week, 1 month and then monthly for 5 months. Fluorescein angiography and optical coherence tomography were performed at 1 month and 3 months after each injection. RESULTS: The mean change in best-corrected visual acuity was -0.06 +/- 0.3 logMAR in 1.25 mg and -0.07 +/- 0.34 in 2.5 mg groups in 3 months (P = 0.9) and -0.06 +/- 0.27 logMAR in 1.25 mg and -0.09 +/- 0.28 in 2.5 mg groups in 5 months (P = 0.6). There was no significant difference in visual acuity between the two groups at any time point during the study. The mean change in foveal thickness was -49 +/- 36 micro in 1.25 mg and -65 +/- 31 micro in 2.5 mg group (P = 0.6). In 2.5 mg group, three cases of vitreous reaction and one case of massive subretinal hemorrhage were observed. CONCLUSION: Intravitreal injection of 2.5 mg bevacizumab has the same efficacy as 1.25 mg, but may be associated with a higher rate of adverse events. PMID- 19287289 TI - First versus second eye intravitreal ranibizumab therapy for wet AMD. AB - PURPOSE: To evaluate the short-term visual outcomes after intravitreal ranibizumab for wet age-related macular degeneration, when used in first eyes (good vision in the untreated eye) compared with second eyes (significant visual impairment in the untreated eye). METHODS: Seventy-five consecutive patients who received intravitreal ranibizumab injection were divided into Group A, comprising 35 first eyed patients and Group B, comprising 40 second eyes. Visual acuity and contrast sensitivity was compared before treatment, and 3 months after the 3rd injection. Results were compared at 95% confidence interval. RESULTS: Mean pretreatment logMar visual acuity was 0.86 (standard deviation 0.28) in Group A whereas Group B was 0.66 (standard deviation 0.36) (P = 0.007). Posttreatment the mean visual acuity in Group A was 0.63 (standard deviation 0.37) and in Group B was 0.44 (standard deviation 0.33) (P = 0.02). The mean numbers of letters gained per patient were 11.1 (Group A) and 10.6 (Group B). Half of all patients showed significant improvement of visual acuity (> or =15 letters gain). Contrast sensitivity significantly improved in both groups and was usually, but not always, associated with visual gain. CONCLUSION: Second eye patients tend to present to clinical diagnosis at a better visual acuity than first ones and subsequently have better chances for better posttreatment visual acuity. However, both groups have an equal chance of significant visual improvement. PMID- 19287290 TI - Marking of retinal breaks in detached retina with laser photocoagulation before pneumatic retinopexy: a prospective case series. PMID- 19287291 TI - The recent article by Shah et al titled "short-term outcomes of 25-gauge vitrectomy with silicone oil for repair of complicated retinal detachment" (Retina 2008;28:723-728). PMID- 19287292 TI - Article on intraocular lenses (IOLs). PMID- 19287295 TI - Article by Drs Bakri and Ekdawi reporting intravitreal silicone oil droplets after intravitreal injections. PMID- 19287297 TI - What's new in Shock, April 2009? PMID- 19287298 TI - Women exposed to single-dose nevirapine in successive pregnancies: effectiveness and nonnucleoside reverse transcriptase inhibitor resistance. AB - OBJECTIVE: To assess the impact of prior exposure to single-dose nevirapine (sdNVP) on mother-to-child transmission and genotypic resistance in HIV-infected women. DESIGN: Prospective study of 120 women exposed to the HIVNET 012 sdNVP regimen in two successive pregnancies and 240 antiretroviral (ARV)-naive, multiparous women who received sdNVP for the first time. RESULTS: One hundred and eight of 120 and 193 of 240 women returned for a postpartum visit by 6 weeks. HIV 1 was detected in 11.1% (95% confidence interval = 5.9-18.6) of the infants of women previously exposed to sdNVP and 4.2% (95% confidence interval = 1.3-7.0) of those exposed for the first time (P = 0.028). Rates of maternal HIV-1 genotypic resistance at 6 weeks postdelivery were 37.5% and 46.4%, respectively (P = 0.119). Sensitive mutation-specific real-time PCR testing found three of 12 previously exposed women who transmitted HIV-1 to their infants had either K103N or Y181C at baseline compared with one of eight ARV-naive, transmitting women who had Y181C. None of 40 randomly selected nontransmitting women from either group had detectable NVP resistance mutations prior to sdNVP exposure. CONCLUSION: This study shows that effectiveness of sdNVP may be compromised by prior exposure to sdNVP, although the increase in transmission rate after prior exposure could not be explained by the detection of NVP resistance mutations prior to re-exposure as measured both by standard genotyping and highly sensitive allele-specific PCR assays. Furthermore, transmission rates of women with prior exposure were not higher than those reported elsewhere. PMID- 19287299 TI - Weight gain at 3 months of antiretroviral therapy is strongly associated with survival: evidence from two developing countries. AB - BACKGROUND: In developing countries, access to laboratory tests remains limited, and the use of simple tools such as weight to monitor HIV-infected patients treated with antiretroviral therapy should be evaluated. METHODS: Cohort study of 2451 Cambodian and 2618 Kenyan adults who initiated antiretroviral therapy between 2001 and 2007. The prognostic value of weight gain at 3 months of antiretroviral therapy on 3-6 months mortality, and at 6 months on 6-12 months mortality, was investigated using Poisson regression. RESULTS: Mortality rates [95% confidence interval (CI)] between 3 and 6 months of antiretroviral therapy were 9.9 (7.6-12.7) and 13.5 (11.0-16.7) per 100 person-years in Cambodia and Kenya, respectively. At 3 months, among patients with initial body mass index less than or equal to 18.5 kg/m (43% of the study population), mortality rate ratios (95% CI) were 6.3 (3.0-13.1) and 3.4 (1.4-8.3) for those with weight gain less than or equal to 5 and 5-10%, respectively, compared with those with weight gain of more than 10%. At 6 months, weight gain was also predictive of subsequent mortality: mortality rate ratio (95% CI) was 7.3 (4.0-13.3) for those with weight gain less than or equal to 5% compared with those with weight gain of more than 10%. CONCLUSION: Weight gain at 3 months is strongly associated with survival. Poor compliance or undiagnosed opportunistic infections should be investigated in patients with initial body mass index less than or equal to 18.5 and achieving weight gain less than or equal to 10%. PMID- 19287300 TI - Detection of tuberculosis in HIV-infected children using an enzyme-linked immunospot assay. AB - OBJECTIVE: To evaluate an enzyme-linked immunospot assay (ELISPOT) for the diagnosis of tuberculosis (TB) in HIV-infected children with suspected TB and to compare the performance of ELISPOT with the tuberculin skin test (TST). METHODS: Interferon-gamma responses to Mycobacterium tuberculosis-specific antigens were measured by ELISPOT in HIV-infected children with suspected TB. HIV-infected and HIV-uninfected children without TB were taken for comparison. RESULTS: Results were available for 188 children, of whom 139 (74%) were HIV-infected. Of these, 22 were classified as having definite TB: 24 probable TB, 14 possible TB and 128 not having TB. The median (range) age of patients was 20 (10-54.1) months. Median interferon-gamma responses to early-secreted antigenic target-6 and culture filtrate protein-10 were higher in children with definite or probable TB compared with children without TB (P < 0.002). In HIV-infected children with an interpretable ELISPOT result, the ELISPOT was positive in 14/21 (66%) with definite TB. A significantly higher proportion of HIV-infected children with definite or probable TB had a positive ELISPOT compared with a positive TST [25/39 (64%) vs. 10/34 (29%), P = 0.005]. In contrast to TST, results from ELISPOT were not affected by young age or severe immunosuppression. In HIV infected children without active TB disease, 27% had a positive ELISPOT, suggesting latent TB infection. CONCLUSION: ELISPOT is more sensitive than TST for the detection of active TB in HIV-infected children. However, the sensitivity of current ELISPOT assays is not sufficiently high to be used as a rule out test for TB. PMID- 19287301 TI - Multiple T-cell epitopes overlap positively-selected residues in the p1 spacer protein of HIV-1 gag. AB - OBJECTIVES: The p1 region of HIV-1 gag contains the frameshift stem-loop, gag-pol transframe and a protease cleavage site that are crucial for viral assembly, replication and infectivity. Identifying and characterizing CD8+ epitopes that are under host immune selection in this region will help in designing effective vaccines for HIV-1. DESIGN: An approach combining bioinformatical analysis and interferon gamma enzyme-linked immunosorbent spot (ELISPOT) assays is used to identify and characterize the epitopes. Potential human leukocyte antigen (HLA) restricted epitopes were identified by correlating the positively-selected mutations with host HLA alleles. METHODS: ELISPOT analysis with overlapping peptides was used to confirm and characterize the epitopes. RESULTS: Four positively-selected residues were significantly associated with HLA class I alleles, including HLA B*1302 (K4R, P = 0.0008 and I5L, P = 0.0108), A*7401 (S9N, P = 0.0002) and A*30 genotypes (P7S, P = 0.009), suggesting epitopes restricted by these alleles are present in this region. ELISPOT analysis with patient peripheral blood mononuclear cells (PBMCs) identified seven novel epitopes restricted by the 3 alleles. Two types of epitopes were observed in this region based on the ELISPOT responses, Type I: the positively-selected variation does not affect CD8+ T-cell responses; and Type II: the CD8+ T-cell responses are determined by the epitope variants. CONCLUSION: We identified and characterized seven novel CD8+ epitopes in the p1 spacer protein region. Classifying the effects of positively-selected variants on CD8+ T-cell responses will help in designing effective vaccines for HIV-1. PMID- 19287302 TI - Excision of nucleoside analogs in mitochondria by p53 protein. AB - OBJECTIVE: Nucleoside analogs, used against HIV, can be incorporated into a mitochondrial DNA by DNA polymerase gamma. Both the decrease in mitochondrial DNA and increased mutations of mitochondrial DNA may lead to mitochondrial diseases. The tumor suppressor protein p53 exhibits 3' --> 5' exonuclease activity and can provide a proofreading function for DNA polymerases. In the present study, we investigated the ability of p53 to excise incorporated nucleoside analogs from DNA in mitochondria. DESIGN: The functional interaction of p53 and DNA polymerase gamma during the incorporation of nucleoside analog was examined in mitochondrial fractions of p53-null H1299 cells, as the source of DNA polymerase gamma. METHODS: Primer extension reactions were carried out to elucidate the incorporation and removal of nucleoside analogs. RESULTS: The results demonstrate that the excision of incorporated nucleoside analogs in mitochondrial fractions of H1299 cells increased in the presence of purified recombinant p53, or cytoplasmic extracts of large cell carcinoma 2 cells expressing endogenous wild type p53 (but not specifically predepleted extracts) or cytoplasmic extracts of H1299 cells overexpressing wild-type p53, but not exonuclease-deficient mutant p53-R175H. The amount of nucleoside analogs incorporated into the elongated DNA with mitochondrial fractions of human colon carcinoma 116 (HCT116)(p53+/+) cells was lower than that of HCT116(p53-/-) cells. Furthermore, mitochondrion-localized elevation of p53 in HCT116(p53+/+) cells, following the irradiation-stress stimuli, correlates with the reduction in incorporation of nucleoside analogs and wrong nucleotides. CONCLUSION: p53 in mitochondria may functionally interact with DNA polymerase gamma, thus providing a proofreading function during mitochondrial DNA replication for excision of nucleoside analogs and polymerization errors. PMID- 19287303 TI - Effects of long-term testosterone administration in HIV-infected women: a randomized, placebo-controlled trial. AB - OBJECTIVE: Androgen deficiency is common in HIV-infected women. We investigated the long-term effects of transdermal testosterone on body composition, bone mineral density, quality of life, and safety. DESIGN: Twenty-five HIV-infected women with free testosterone below the median (< or =3 pg/ml) of the female normal range were randomized to receive transdermal testosterone (300 microg twice weekly) or identical placebo over 18 months. RESULTS: Women demonstrated low androgen levels (1.3 +/- 0.1 pg/ml) with relatively low weight (22.8 +/- 0.6 kg/m) and low bone mineral density (-0.61 +/- 0.17 SD hip T score) at baseline. No statistically significant differences were seen between the groups at baseline. The discontinuation rate was 16% and did not differ between treatment groups (P = 0.24). Free testosterone by equilibrium dialysis increased over 18 months (7.9 +/- 1.8 vs. 0.3 +/- 0.4 pg/ml; P = 0.002, testosterone vs. placebo). Testosterone was well tolerated and did not affect lipids, liver, or safety indices. Lean mass (1.8 +/- 0.5 vs. 0.8 +/- 0.9 kg; P = 0.04) and BMI (1.6 +/- 0.4 vs. 0.8 +/- 0.6 kg/m; P = 0.03, testosterone vs. placebo) increased in response to testosterone, whereas fat mass remained unchanged. Testosterone increased bone mineral density at the hip (0.01 +/- 0.01 vs. -0.01 +/- 0.01 g/cm; P = 0.02) and trochanter (0.01 +/- 0.01 vs. -0.02 +/- 0.01 g/cm; P = 0.01, testosterone vs. placebo). Testosterone significantly improved depression indices (-6.8 +/- 2.2 vs. -1.9 +/- 3.1; P = 0.02) and problems affecting sexual function (-1.8 +/- 0.8 vs. 0.5 +/- 0.5; P = 0.01, testosterone vs. placebo). CONCLUSION: Long-term testosterone administration was well tolerated in HIV-infected women and resulted in significant improvements in body composition, bone mineral density, and quality of life indices. Further evaluation of the safety and efficacy of testosterone use among HIV-infected women is warranted. PMID- 19287304 TI - Lapatinib plus capecitabine resolved human epidermal growth factor receptor 2 positive brain metastases. AB - Brain metastases affect 25%-30% of women with human epidermal growth factor receptor 2 (HER2)-positive metastatic breast cancer and are associated with a high burden of disease and poor prognosis. A 55-year-old woman presented with HER2-positive, hormone receptor-positive, locally advanced infiltrating ductal carcinoma. She received 4 cycles of neoadjuvant docetaxel (75 mg/m) plus trastuzumab (6 mg/kg) on a 21-day cycle, resulting in complete pathologic response at the time of surgery. Trastuzumab (6 mg/kg every 21 days) plus anastrozole (1 mg/d) was continued for 1 year. Two years later, the patient progressed with pulmonary nodules and a large pleural effusion. Computed tomography and positron emission tomography revealed multiple lesions in the liver and thoracic spine but no evidence of brain metastases. The patient received weekly trastuzumab (2 mg/kg), paclitaxel (80 mg/m), and carboplatin (area under the curve 2) for 6 months; her symptoms resolved and her disease stabilized. Seven months later, she developed diplopia and gait difficulties, and magnetic resonance imaging revealed multiple brain lesions. Whole-brain radiotherapy (30 Gy in 10 fractions) was delivered with excellent clinical results. The patient remained progression free without symptoms for approximately 3 months. When she developed central nervous system symptoms, she was treated with lapatinib (1250 mg/d continuously) plus capecitabine (2000 mg/m given on days 1-14 of a 21-day cycle). Four months later, a brain computed tomography performed shortly before her death from progressive systemic disease revealed near complete resolution of brain metastases. Lapatinib plus capecitabine seems to have clinical activity in HER2-positive brain metastases. PMID- 19287305 TI - Multiple hepatic arterial injections of recombinant adenovirus p53 and 5 fluorouracil after transcatheter arterial chemoembolization for unresectable hepatocellular carcinoma: a pilot phase II trial. AB - This pilot phase II study was designed to determine the efficacy, toxicities, and biological activity of multiple hepatic arterial injections of recombinant adenovirus p53 (rAd-p53) and 5-fluorouracil (5-FU) after transcatheter arterial chemoembolization (TACE) when compared with TACE alone in patients with unresectable hepatocellular carcinoma (HCC). Forty-six patients with unresectable HCC were randomized in either group 1 [23 patients, multiple hepatic arterial injections of Ad-p53 (1x10 viral particles) and 5-FU (500-750 mg), after TACE] or group 2 (23 patients, TACE alone). In group 1, the number of Ad-p53/5-FU courses administered was 166 (median 7, range 3-12). In group 2, the number of TACE courses administered was 47 (median 2, range 1-3). Partial response and stable disease were 69.5% in group 1 and 65.2% in group 2. Times to progression were 9.6 months (range 2.1-21.7) in group 1 and 8.3 months (range 2.1-16.8) in group 2. Overall survivals were 12.8 months (range 2.7-26.2) in group 1 and 10.4 months (range 2.7-22.5) in group 2. Toxicities in both groups were generally mild and reversible. The most common Ad-p53-related toxicity was a transient fever. Specific p53 transgene expression was detected using reverse-transcriptase polymerase chain reaction in biopsied tumor tissues. Distribution studies revealed that the vector was detected in the plasma, but rarely in the gargle and urine. This study shows that multiple hepatic arterial injections of Ad-p53 and 5 FU after TACE can be active and safe as a treatment for patients with unresectable HCC. PMID- 19287306 TI - Administration of reduced glutathione in FOLFOX4 adjuvant treatment for colorectal cancer: effect on oxaliplatin pharmacokinetics, Pt-DNA adduct formation, and neurotoxicity. AB - Oxaliplatin is a promising drug for cancer therapy and the oxaliplatin/5 fluorouracil/leucovorin (FOLFOX) regimen has become the standard adjuvant treatment for colorectal cancer. However, the oxaliplatin-induced neurotoxicity still represents a clinical problem leading to a discontinuation of the therapy. Many strategies have been proposed in order to manage the neurotoxicity, but their effect on antitumoral efficacy is still unclear. In this study, we investigated the effect of reduced glutathione administration on neurotoxicity, oxaliplatin pharmacokinetics, and platinum-DNA (Pt-DNA) adduct formation in patients affected by colorectal cancer treated with FOLFOX4 adjuvant regimen. Twenty-seven patients were randomized to receive GSH 1500 mg/m or saline solution before oxaliplatin infusion. Evaluation of neurotoxicity, pharmacokinetics of plasmatic total and ultrafiltered Pt, and determination of Pt-DNA adduct formation on white blood cells was performed during the 5th, 9th, and 12th cycles. At the end of all cycles of therapy, the patients in the GSH arm showed a statistically significant reduction of neurotoxicity (P=0.0037) compared with the placebo arm. There were no significant differences in the main pharmacokinetic parameters between the two arms except a lower area under the plasma concentration-time curve and a smaller apparent steady-state volume of distribution (Vss) when GSH was coadministered. This difference can be explained by the natural function of GSH in the detoxification of oxaliplatin and by its ability to remove the Pt bound to plasma proteins. The determination of Pt-DNA adduct formation shows no statistically significant differences between the two arms. In conclusion, this study indicates that coadministration of GSH is an effective strategy to reduce the oxaliplatin-induced neurotoxicity without impairing neither the pharmacokinetics of oxaliplatin, nor the Pt-DNA adduct formation. PMID- 19287308 TI - Adult cases of leukoencephalopathy, cerebral calcifications, and cysts: expanding the spectrum of the disorder. AB - Leukoencephalopathy with cerebral calcifications and cysts (LCC) was first reported in children who developed cognitive decline and variable extrapyramidal, cerebellar, and pyramidal signs, with or without seizures. Leukoencephalopathy with cerebral calcifications and cysts is characterized by progressive formation of brain cysts that can generate a mass effect simulating a neoplasm. Retinal changes that overlap with Coats disease, a microangiopathy with retinal telangiectasias and exudates, may also occur. We and others have reported LCC cases in adults. Neuroimaging shows diffuse leukoencephalopathy, multifocal calcifications especially of deep gray and white matter, multifocal enhancement, and variably sized cysts that may require surgical decompression. Biopsies adjacent to cysts have shown angiomatous and/or severely hyalinized blood vessels surrounded by myelin loss and gliosis, calcifications, and Rosenthal fibers. We report 2 additional adult-onset cases of LCC. Case 1 is a 40-year-old man who developed neurological symptoms and cirrhosis and died of acute gastrointestinal bleeding; he had numerous retinal microinfarcts at autopsy. Case 2 is a 55-year old woman who was found by chance to have LCC; one and a half years later, her course remains benign. These cases expand the spectrum of adult-onset LCC, the etiology of which is unknown. PMID- 19287309 TI - Caveolin 1 expression independently predicts shorter survival in oligodendrogliomas. AB - Caveolin 1 (cav-1) is the basic component of the flask-shaped membrane microdomains known as caveolae that are involved in various cell functions. Caveolin 1 can be overexpressed in tumors,suggesting a proneoplastic role, or it can be downregulated. We previously reported that cav-1 expression increases with tumor grade in astrocytomas. Here, we studied cav-1 immunoreactivity in brain umors with an oligodendroglial component to determine the prognostic value of cav 1 expression and to correlate it with 1p/19q deletions. Fifty-four oligodendrogliomas, 26 mixed oligoastrocytomas,and 7 glioblastomas with an oligodendroglial component were assessed for cav-1 expression by immunohistochemistry and for 1p/19q status by fluorescence in situ hybridization. Caveolin-1 was detected in a minority of cases (22%) and was associated mostly with Grade III mixed oligoastrocytomas and glioblastomas with anoligodendroglial component; cav-1 expression was significantly correlated with the absence of a 1p/19q deletion (p = 0.0002). In the 63 cases in which survival data were available, cav-1 expression was also significantly associated with shorter survivals, whereas 1p/19q deletion was associated with longer survivals. Among high-grade tumors, cav-1 expression was the only factor that retained a statistical significance after multivariate analysis for the prediction ofa short survival (p G 0.015). These data are the first evidence that cav-1 immunohistochemistry is an independent prognostic marker in tumors with an oligodendroglial component regardless of the 1p/19q status. PMID- 19287307 TI - EUROASPIRE III: a survey on the lifestyle, risk factors and use of cardioprotective drug therapies in coronary patients from 22 European countries. AB - AIM: The aim of the European Action on Secondary and Primary Prevention by Intervention to Reduce Events III (EUROASPIRE III) survey was to determine whether the Joint European Societies' guidelines on cardiovascular prevention are being followed in everyday clinical practice and to describe the lifestyle, risk factor and therapeutic management in patients with coronary heart disease (CHD) in Europe. METHODS: The EUROASPIRE III survey was carried out in 2006-2007 in 76 centres from selected geographical areas in 22 countries in Europe. Consecutive patients, with a clinical diagnosis of CHD, were identified retrospectively and then followed up, interviewed and examined at least 6 months after their coronary event. RESULTS: Thirteen thousand nine hundred and thirty-five medical records (27% women) were reviewed and 8966 patients were interviewed. At interview, 17% of patients smoked cigarettes, 35% were obese and 53% centrally obese, 56% had a blood pressure >or=140/90 mmHg (>or=130/80 in people with diabetes mellitus), 51% had a serum total cholesterol >or=4.5 mmol/l and 25% reported a history of diabetes of whom 10% had a fasting plasma glucose less than 6.1 mmol/l and 35% a glycated haemoglobin A1c less than 6.5%. The use of cardioprotective medication was: antiplatelets 91%; beta-blockers 80%; angiotensin-converting enzyme inhibitors/angiotensin-receptor blockers 71%; calcium channel blockers 25% and statins 78%. CONCLUSION: The EUROASPIRE III survey shows that large proportions of coronary patients do not achieve the lifestyle, risk factor and therapeutic targets for cardiovascular disease prevention. Wide variations in risk factor prevalences and the use of cardioprotective drug therapies exist between countries. There is still considerable potential throughout Europe to raise standards of preventive care in order to reduce the risk of recurrent disease and death in patients with CHD. PMID- 19287310 TI - Cerebrovascular smooth muscle actin is increased in nondemented subjects with frequent senile plaques at autopsy: implications for the pathogenesis of Alzheimer disease. AB - We previously found that vascular smooth muscle actin (SMA) is reduced in the brains of patients with late stage Alzheimer disease (AD) compared with brains of nondemented, neuropathologically normal subjects. To assess the pathogenetic significance and disease specificity of this finding, we studied 3 additional patient groups: nondemented subjects without significant AD type pathology ("Normal"; n = 20), nondemented subjects with frequent senile plaques at autopsy ("Preclinical AD"; n = 20), and subjects with frontotemporal dementia ("FTD"; n = 10). The groups were matched for sex and age with those previously reported; SMA immunohistochemistry and image analysis were performed as previously described. Surprisingly, SMA expression in arachnoid, cerebral cortex, and white matter arterioles was greater in the Preclinical AD group than in the Normal and FTD groups. The plaques were not associated with amyloid angiopathy or other vascular disease in this group. Smooth muscle actin expression in the brains of the Normal group was intermediate between the Preclinical AD and FTD groups. All 3 groups exhibited much greater SMA expression than in our previous report. The presence of frequent plaques and increased arteriolar SMA expression in the brains of nondemented subjects suggest that increased SMA expression might represent a physiological response to neurodegeneration that could prevent or delay overt expression dementia in AD. PMID- 19287311 TI - Addressing a folate imbalance in fetal cerebrospinal fluid can decrease the incidence of congenital hydrocephalus. AB - Fetal-onset hydrocephalus (HC), which affects between 1:500 and 1:5000 live human births, results from unequal production and drainage of cerebrospinal fluid (CSF) and is associated with abnormal development of the cerebral cortex leading to severe neurological deficits. We previously found that in the hydrocephalic Texas rat, the CSF of affected fetuses induced a cell cycle arrest in neural progenitor cells. Here, we show that alterations in folate metabolism in the CSF of the developing cerebrum are likely responsible for this effect. We identified 3 folate enzymes in the CSF and demonstrate that low levels of one of these, 10 formyltetrahydrofolate dehydrogenase, are associated with HC in the hydrocephalic Texas rat. Therefore, we tested whether supplementation with specific folate species would improve developmental outcome. After daily administration of a combination of tetrahydrofolic and 5-formyltetrahydrofolic acids to pregnant dams, there was a significant reduction in the incidence of HC and improved brain development. By contrast, supplementation with folic acid increased the incidence of congenital HC in this model. These results indicate the complexities of folate metabolism in the developing brain and suggest that folate imbalance leading to HC in the hydrocephalic Texas rat fetuses can be treated with maternal folate supplementation using specific folate metabolites and combinations thereof. PMID- 19287312 TI - Interleukin-10 and interleukin-10 receptor-I are upregulated in glial cells after an excitotoxic injury to the postnatal rat brain. AB - Inflammation is an important determinant of the severity and outcome of central nervous system injury. The endogenous anti-inflammatory cytokine interleukin-10 (IL-10) is upregulated in the injured adult central nervous system where it controls and terminates inflammatory processes. The developing brain, however, displays differences in susceptibility to insults and in associated inflammatory responses from the adult brain; the anatomic and temporal patterns of injury induced IL-10 expression in the immature brain after excitotoxic injury are unknown. We analyzed the spaciotemporal gene and protein expression of IL-10 and its receptor (IL-10RI) in N-methyl-d-aspartate-induced excitotoxic injury in 9 day-old and control rats using quantitative reverse transcriptase polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemistry. In noninjected control brains, both molecules were expressed mainly in white matter on glial cells and blood vessels; IL-10 was also observed on blood vessels in gray matter and in glial fibrillary acidic protein-positive processes in the hippocampus and near leptomeningeal and ventricle surfaces. In N-methyl-d aspartate-injected brains, IL-10 gene and protein expression were maximal at 72 hours postinjection; IL-10RI gene and protein expression peaked at 48 hours postinjection. Interleukin-10 and IL-10RI expression in injured areas was mainly found in reactive astrocytes and in microglia/macrophages. The expression patterns of IL-10 and IL-10R suggest possible developmental roles, and their upregulation after injury suggests that this expression may have anti inflammatory effects in distinct anatomic sites in the immature brain. PMID- 19287313 TI - Sarcolemmal neuronal nitric oxide synthase defect in limb-girdle muscular dystrophy: an adverse modulating factor in the disease course? AB - Reduction of neuronal nitric oxide synthase (nNOS) has been associated with the pathogenesis and clinical expression of inherited myopathies. To determine whether a defect in nNOS might be an adverse modulating factor in the course of limb-girdle muscular dystrophy, we investigated cytosolic and sarcolemmal nNOS expression in muscle biopsies from 32 patients with 7 forms of limb-girdle muscular dystrophy. Primary calpainopathy, dysferlinopathy, and caveolinopathy biopsies showed normal levels of cytosolic nNOS and preserved sarcolemmal nNOS immunoreactivity. By contrast, the cytosolic nNOS levels in sarcoglycanopathy muscles were variably reduced. Sarcolemmal nNOS immunoreactivity varied from absent to reduced, depending on the integrity of the sarcoglycan complex. In muscles with loss of the entire sarcoglycan complex, sarcolemmal nNOS was absent; it otherwise depended on the specific sarcoglycan gene and type of mutation. The integrity of the entire sarcoglycan complex is, therefore, essential for the stabilization of nNOS to the sarcolemma. Absence of sarcolemmal nNOS in sarcoglycanopathy muscle was always associated with severe muscular dystrophy and sometimes with dilated cardiomyopathy, supporting the hypothesis that nNOS defect might contribute to skeletal and cardiac muscle disease progression. These results emphasize the value of nNOS immunohistochemical analysis in limb-girdle muscular dystrophy and provide additional insights for future therapeutic interventions in these disorders. PMID- 19287314 TI - Metabotropic glutamate receptor II in the brains of Parkinsonian patients. AB - Modulation of basal ganglia group II metabotropic glutamate receptors (mGluR2/3) is a potential therapeutic alternative to levodopa in Parkinson disease (PD). We used receptor-binding autoradiography of the mGluR2/3-selective radioligand [H]LY341495 in postmortem brain specimens from PD patients (n = 14) and controls (n=11) to investigate possible contributions of changes in ligand binding of this receptor to levodopa-associated motor complications experienced premortem in PD patients. The PD patients included those with and without histories of dyskinesias and those with and without "wearing off," which is defined as a reduced period of benefit from levodopa. Specific binding of [H]LY341495 to mGluR2/3 in the basal ganglia was higher in the caudate nucleus than the putamen and lower by approximately half in the external and internal globus pallidus (GPi) in controls. [H]LY341495-specific binding was reduced in the caudate and GPi in patients without wearing-off (-22% caudate, -30% GPi), compared with controls and with patients who had experienced wearing-off; there were no differences among PD patients with or without dyskinesias. These data suggest that an adaptive downregulation of mGluR2/3 in PD patients without wearing-off may compensate for increased glutamate. They indicate a key role for mGluR2/3 in control of movement and the potential for mGluR2/3-targeted drugs in the management of wearing-off fluctuations in PD. PMID- 19287315 TI - Mitochondrial alterations in transgenic mice with an H46R mutant Cu/Zn superoxide dismutase gene. AB - We examined morphological alterations in the mitochondria in the spinal cord of H46R mutant Cu/Zn superoxide dismutase transgenic mice; these mice serve as a model for human familial amyotrophic lateral sclerosis. The disease in the mice is characterized by initial muscle weakness and atrophy in the legs, very long clinical courses, and widespread pathological changes of the spinal cord that extend beyond the motor system and includes many neuropil aggregates that lack vacuoles. At the preclinical stage, we found alterations in the mitochondrial cristae that included focal electron-dense changes, whorled membranous and electron-dense amorphous structures, and outward projections of outer and inner membranes predominantly in proximal axons. At the overt disease stage, these mitochondrial alterations were more frequent and were also found in somata, dendrites, presynaptic terminals, and astrocyte cytoplasm. By immunoelectron microscopy, no accumulations of Cu/Zn superoxide dismutase- or ubiquitin-positive immunogold particles were observed in either normal-appearing or abnormal mitochondria. These findings suggest that predominant mitochondrial alterations in the proximal axons begin in the preclinical stage and may be involved in the pathogenetic mechanisms of motor neuron degeneration in these transgenic mice via disruption of the axonal transport of substrates necessary for neuronal viability; this disruption may lead to motor neuron death. PMID- 19287316 TI - Increased reelin promoter methylation is associated with granule cell dispersion in human temporal lobe epilepsy. AB - Mesial temporal sclerosis (MTS) is the most common lesion in chronic, intractable temporal lobe epilepsies (TLE) and characterized by segmental neuronal cell loss in major hippocampal segments. Another histopathological hallmark includes granule cell dispersion (GCD), an architectural disturbance of the dentate gyrus encountered in approximately 50% of patients with mesial temporal sclerosis. Reelin, which plays a key role during hippocampal development and maintenance of laminar organization, is synthesized and released by Cajal-Retzius cells of the dentate molecular layer, and previous studies have shown that Reelin transcript levels are downregulated in human temporal lobe epilepsies specimens. To investigate whether epigenetic silencing by Reelin promoter methylation may be an underlying pathogenetic mechanism of GCD, DNA was harvested from 3 microdissected hippocampal subregions (i.e. molecular and granule cell layers of the dentate gyrus and presubiculum) from 8 MTS specimens with GCD, 5 TLE samples without GCD, and 3 autopsy controls. Promoter methylation was analyzed after bisulfite treatment, cloning, and direct sequencing; immunohistochemistry was performed to identify Cajal-Retzius cells. Reelin promoter methylation was found to be greater in TLE specimens than in controls; promoter methylation correlated with GCD among TLE specimens (p < 0.0002). No other clinical or histopathological parameter (i.e. sex, age, seizure duration, medication or extent, of MTS) correlated with promoter methylation. These data support a compromised Reelin-signaling pathway and identify promoter methylation as an epigenetic mechanism in the pathogenesis of TLE. PMID- 19287318 TI - Enhanced fear responses in mice treated with anabolic androgenic steroids. AB - Corticolimbic neurons express neurosteroid biosynthesis, which is altered during anabolic androgenic steroid (AAS) treatment. The brain circuits and neurons that underlie the behavioral deficits found after AAS treatment remain undefined. We studied the effects of testosterone propionate (testosterone) on fear conditioning responses and in primary output corticolimbic neurons on 5alpha reductase-type-I and 3alpha-hydroxysteroid-dehydrogenase expression. Testosterone fails to change cued fear responses although it induces excessive contextual fear associated with corticolimbic 5alpha-reductase-type-I mRNA expression downregulation in the prefrontal cortex, hippocampus, and basolateral amygdala glutamatergic neurons. Increased fear responses are abolished by normalizing corticolimbic allopregnanolone levels with allopregnanolone treatment (8 micromol/kg) or selective brain steroidogenic stimulants, including S norfluoxetine (1.8 micromol/kg). Agents that increase corticolimbic allopregnanolone levels may be beneficial in treating AAS users. PMID- 19287319 TI - Afferent islands are larger than mu-opioid receptor patch in striatum of rat pups. AB - Dopamine afferent islands were observed in rodent caudate-putamen only during development, whereas patches with intense mu-opioid receptor (MOR) immunoreactivity were seen throughout the life. We performed direct comparison between MOR patches and dopamine islands in the caudate-putamen of rat pups, by double immunofluorescence labeling for MOR and tyrosine hydroxylase. MOR patches were included in dopamine islands at postnatal day (P) 0 to P8, although the patches occupied the same region as the islands at P12-16. Furthermore, the regions of glutamatergic afferents with intense vesicular glutamate transporter 1 and vesicular glutamate transporter 2 immunoreactivities well corresponded to those of dopamine islands at P4. These results suggest that the striatal 'afferent islands' are larger than MOR patches in the early postnatal life. PMID- 19287320 TI - MPP+ impairs autophagic clearance of alpha-synuclein by impairing the activity of dynein. AB - Increasing evidence suggests that dynein has an important role in the clearance of misfolded proteins by autophagy. Here we show that treatment of cells with 1 methyl-4-phenylpyridinium (MPP) cause alpha-synuclein overexpression and aggregation, leading to the accumulation of autophagic vacuoles and the recruitment of LC3-II to these vacuoles in the cytoplasm. After MPP treatment, dynein expression decreased and was mainly aggregated at the periphery of cytoplasm and lost its colocalization with alpha-synuclein and lamp1, indicating that dynein lost its function in the aggresome formation and failed to return autophagosome and lysosomes to the center of the cell for degradation. We consider that dynein plays an important role in the autophagic clearance of aggregate-prone proteins. PMID- 19287321 TI - Joint attention helps infants learn new words: event-related potential evidence. AB - This study investigated the role of joint attention in infants' word learning. Infants aged 18-21 months were taught new words in two social contexts, joint attention (eye contact, positive tone of voice) or non-joint attention (no eye contact, neutral tone of voice). Event-related potentials were measured as the infants saw objects either congruent or incongruent with the taught words. For both social contexts, an early negativity was observed for the congruent condition, reflecting a phonological-lexical priming effect between objects and the taught words. In addition, for the joint attention, the incongruent condition elicited a late, widely distributed negativity, attributed to semantic integration difficulties. Thus, social cues have an impact on how words are learned and represented in a child's mental lexicon. PMID- 19287322 TI - The role of the posterior superior temporal cortex in sentence comprehension. AB - The study investigates to what extent the posterior superior temporal cortex is involved in processing complex sentences. Using functional MRI, we show that hierarchically structured sentences activate the superior temporal cortex bilaterally to greater extent than sentences with a linear structure. The activation in the left hemisphere comprises the superior temporal gyrus and sulcus, whereas the activation in the right hemisphere is confined to the superior temporal sulcus. As earlier studies using similar syntactic structures in semantic-free grammars did not show activation in the superior temporal cortex but instead only in the prefrontal cortex, we conclude that the role of the posterior superior temporal cortex is to integrate lexical-semantic and syntactic information during sentence comprehension. PMID- 19287323 TI - Transoral approach and its superior extensions to the craniovertebral junction malformations: surgical strategies and results. AB - OBJECTIVE: To review our experience with the surgical management of craniovertebral junction malformations, focusing on the selection of surgical approach, management of the associated Chiari malformation, and postoperative instability. METHODS: During a 7-year period (May 2000-May 2007), 34 patients with a mean age of 55 years (age range, 32-75 years) underwent transoral surgery for fixed or nearly fixed ventral compression at the craniovertebral junction caused by basilar invagination and/or atlantoaxial dislocation. Chiari malformation was detected in 13 patients. The most common presenting signs were motor deficits (88%), followed by sensory loss (35%). All patients but one who had posterior stabilization performed elsewhere underwent single-stage anterior decompression and posterior occipitocervical fixation. Adjuncts to the transoral approach were tailored to the local anatomy (severity of basilar invagination, extent of mandibular excursion) found in each patient. Posterior fossa decompression was performed in 3 patients with Chiari malformation. RESULTS: Thirty-one patients were alive at the time of the last follow-up evaluation (average, 3.7 years; range, 0.5-7.5 years). Of the 28 surviving patients admitted with preoperative motor impairment, 24 patients (86%) improved at least 1 Nurick grade, whereas the grade did not change in 4 (14%) patients. There were 2 (6%) perioperative deaths, and 1 other patient died subsequently of causes unrelated to surgery. Surgical morbidity was 18% and included dural laceration, cerebrospinal fluid leak with meningitis, malocclusion, oral wound dehiscence, and occipital wound infection. Delayed instability occurred in 1 patient because of cranial settling of the C2 vertebral body. CONCLUSION: Successful decompression of the abnormal craniovertebral junction requires extensive preoperative evaluation, appropriate tailoring of the operative approach, and an adequate learning curve. Transmaxillary approaches are useful adjuncts to the transoral approach in patients with severe basilar invagination or in cases of limited jaw mobility. Anterior decompression has been proven effective in relieving obstruction of the subarachnoid space at the foramen magnum in most patients with associated Chiari malformation. PMID- 19287324 TI - Endonasal versus supraorbital keyhole removal of craniopharyngiomas and tuberculum sellae meningiomas. AB - OBJECTIVE: Endonasal and supraorbital "eyebrow" craniotomies are increasingly being used to remove craniopharyngiomas and tuberculum sellae meningiomas. Herein, we assess the relative advantages, disadvantages, and selection criteria of these 2 keyhole approaches. METHODS: All consecutive patients who had endonasal or supraorbital removal of a craniopharyngioma or tuberculum sellae meningioma were analyzed. RESULTS: Of 43 patients, 22 had a craniopharyngioma (18 endonasal, 4 supraorbital), and 21 had a meningioma (12 endonasal, 7 supraorbital, 2 both routes); 33% had prior surgery. Craniopharyngiomas were primarily retrochiasmal in location in 78% of endonasal cases versus 25% of supraorbital cases (P = 0.08). Meningiomas were larger when approached by the supraorbital route versus the endonasal route (33 +/- 10 versus 25 +/- 8 mm, respectively; P = 0.008). Endoscopy was used in 84% of endonasal approaches and in 31% of supraorbital approaches (P = 0.001). Of patients having first-time surgery for a craniopharyngioma (n = 14) or meningioma (n = 15), total/near total removal was achieved in 83% and 80% of patients by the endonasal route and in 50% and 80% of patients by the supraorbital route, respectively. Vision improved in 87% and 70% of patients who had surgery by an endonasal versus supraorbital route, respectively (P = 0.3). Visual deterioration occurred in 2 patients with meningiomas, 1 by endonasal (7%), and 1 by supraorbital (11%) removal. The endonasal approach was associated with a higher rate of postoperative cerebrospinal fluid leaks (16 versus 0%; P = 0.3), 4 of 5 of which occurred in patients with meningioma. CONCLUSION: The endonasal route is preferred for removal of most retrochiasmal craniopharyngiomas, whereas the supraorbital route is recommended for meningiomas larger than 30 to 35 mm or with growth beyond the supraclinoid carotid arteries. For smaller midline tumors, either approach can be used, depending on surgeon experience and tumor anatomy. Compared with traditional craniotomies, the major limitation of both approaches is a narrow surgical corridor. The endonasal approach has the added challenges of restricted lateral suprasellar access, a greater need for endoscopy, and a more demanding cranial base repair. PMID- 19287325 TI - Surgical considerations in the management of falcotentorial meningiomas: advantages of the bilateral occipital transtentorial/transfalcine craniotomy for large tumors. AB - OBJECTIVE: Meningiomas arising from the falcotentorial junction are relatively rare, and the description of the surgical nuances in approaching these tumors is limited. We describe our surgical management of these lesions in detail. METHODS: From 2001 to 2005, 9 patients underwent operation for meningiomas arising from the falcotentorial junction, with some extending to and/or invading the torcula. All patients were assessed preoperatively with magnetic resonance neuroimaging and cerebral angiography. Furthermore, preoperative embolization was attempted in all cases. A supratentorial/infratentorial torcular craniotomy technique was used in all but 1 of these cases. RESULTS: The average dimensions of the falcotentorial meningiomas were 5.1 x 4.4 x 4.2 cm. The angiograms revealed that these tumors were fed by branches of the internal carotid artery, choroidal arteries, branches of the meningohypophyseal trunk, and branches of the posterior cerebral artery. Preoperative embolization was achieved in only 2 patients. Five patients had gross total resection (Simpson grade 1), and 4 had subtotal resection (Simpson grade 4). Two of the tumors (22%) recurred during a mean follow-up period of 49 months (range, 17-88 months). The most common complication after surgery was cortical blindness, but all postoperative visual deficits had fully recovered at the last follow-up evaluation within several months. CONCLUSION: An excellent outcome can be expected with detailed preoperative neuroimaging and knowledge of the nuances of the surgical technique that we describe in detail in this article. PMID- 19287326 TI - Endovascular treatment of middle cerebral artery aneurysms with detachable coils: angiographic and clinical outcomes in 115 consecutive patients. AB - OBJECTIVE: Because of their anatomic configuration, middle cerebral artery (MCA) aneurysms are most often treated with surgical clipping. However, endovascular coil embolization of these aneurysms is an increasingly used alternative. We retrospectively reviewed the anatomic and clinical outcomes of patients with MCA aneurysms who underwent endovascular treatment at our institution. METHODS: One hundred fifteen MCA aneurysms in 115 patients (mean age, 55.1 years) were treated by an endovascular technique from April 1990 to March 2007. Forty-eight patients (42%) presented with acute subarachnoid hemorrhage, and 67 patients (58%) had unruptured aneurysms. Fifty-three aneurysms (46%) were small with a small neck, 28 (24%) were small with a wide neck, 22 (19%) were large, and 12 (11%) were giant. RESULTS: Angiographic results immediately after embolization showed complete occlusion in 53 aneurysms (46%), a neck remnant in 51 (44%), and incomplete occlusion in 3 (3%). Because of anatomic difficulties, we could not embolize 8 aneurysms (7%). Thirteen patients underwent combined treatment that included endovascular and extracranial-intracranial bypass surgery. Morbidity and mortality rates were 6.9% (8 patients) and 3% (3 patients), respectively. Procedure-related complications were encountered in 10 patients (9%). Seventy patients had long-term follow- up angiograms. Seven aneurysms (10%) were recanalized; all were large or giant. One partially embolized large aneurysm ruptured 13 months after embolization. CONCLUSION: In this series, endovascular coil embolization of MCA aneurysms has morbidity and mortality rates comparable to those of conventional surgical clipping. Combined treatment of endovascular and bypass surgery can successfully treat large or giant complex fusiform MCA aneurysms. PMID- 19287327 TI - Comparative study of decompressive craniectomy after mass lesion evacuation in severe head injury. AB - OBJECTIVE: This study was conducted to evaluate outcome after decompressive craniectomy (DC) in the setting of mass evacuation with or without intracranial pressure (ICP) monitoring. METHODS: Over a 48-month period (March 2000 to March 2004), 54 of 967 consecutive head injury patients underwent DC for evacuation of a mass lesion. DC was performed without ICP monitoring in 27 patients who required urgent decompression (group A) and in 27 patients who did not require urgent surgery and who had their ICP monitored for 1 to 14 days before surgery (group B). RESULTS: In group A, the mean Glasgow Coma Scale score was 6.0; 80% had computed tomographic evidence of a shift greater than 5 mm; and 25 patients underwent DC immediately after resuscitation. In group B, the mean Glasgow Coma Scale score was 7.3; 40% had computed tomographic evidence of shift; and 75% underwent DC 24 hours or longer after presentation. Overall, 22 patients died (12 in group A and 10 in group B), 11 remained vegetative or severely disabled (3 in group A and 8 in group B), and 19 had good recovery (11 in group A and 8 in group B). Two patients were lost to follow-up. In 18 group B patients with ICP greater than 20 mm Hg before mass evacuation, ICP dropped an average of 13 mm Hg (P < 0.001). A mass lesion greater than 50 mL (odds ratio [OR], 2.86; 95% confidence interval [CI], 1.04-7.89) and evidence of low attenuation on computed tomography before (OR, 3.3; 95% CI, 1.1-10.3) or after (OR, 2.92; 95% CI, 1.02-8.34) DC were predictors of death. A good outcome occurred in 42% of patients with and in 63% of patients without delayed traumatic injury (OR, 0.3; 95% CI, 0.1-1.1). Outcome was favorable in 78.6% of patients who had no ICP monitoring before DC versus 47.1% of patients with ICP monitoring (OR, 0.2; 95% CI, 0.1-1.2). CONCLUSION: In this study, mortality after DC for mass lesion was greater than expected, and outcome did not differ between patients with or without ICP monitoring. PMID- 19287328 TI - Factors associated with aspirin resistance in patients premedicated with aspirin and clopidogrel for endovascular neurosurgery. AB - OBJECTIVE: Antiplatelet therapy is critical to endovascular neurosurgical procedures. Some patients are aspirin-resistant nonresponders. We reviewed our endovascular neurosurgery patients who were premedicated with aspirin and clopidogrel and identified nonresponders to aspirin. Factors associated with aspirin resistance were determined. METHODS: Consecutive endovascular neurosurgery patients were identified who were treated by the senior author (BLH) from December 2006 to October 2007 and who were premedicated with aspirin (325 mg) and clopidogrel (75 mg) for 7 days before the procedure. We retrospectively reviewed values from the platelet function analyzer-100 test (Dade-Behring, Deerfield, IL) from 1 day before the procedures. The following factors were evaluated for association with aspirin drug resistance: age, sex, body mass index, and smoking history; patients with hypertension, diabetes, coronary artery disease/ peripheral vascular disease, or hypercholesterolemia; disease pathology (aneurysm, intracranial stenosis, or extracranial stenosis); patients taking statins, beta-blockers, angiotensin-converting enzyme inhibitors/angiotensin receptor blockers, or antidepressants; and white blood cell count, hemoglobin, hematocrit, and platelet levels. A stepwise logistic model selection was used to select important factors and their interactions. RESULTS: Eighty-one consecutive patients with the following interventions were included in the study: 35 aneurysm coilings (43%), 21 stent-assisted aneurysm coilings (26%), 13 carotid stent and angioplasties (16%), 7 intracranial stents and angioplasties (9%), and 5 extracranial vertebral artery stents and angioplasties (6%). Seventeen patients (21%) were nonresponders to aspirin. After model selection, the only factor associated with aspirin resistance was not taking an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker (P = 0.0348; odds ratio, 0.214; 95% confidence interval, 0.051-0.896). CONCLUSION: Twenty-one percent of patients premedicated with aspirin and clopidogrel dual therapy for 7 days before endovascular neurosurgical procedures were nonresponders to aspirin. Patients not taking an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker may be at higher risk for aspirin drug resistance. PMID- 19287329 TI - The role of epoxide hydrolase Y113H gene variant in pancreatic diseases. AB - OBJECTIVES: Chronic pancreatitis (CP) and pancreatic adenocarcinoma (pCA) are associated with risk factors such as alcohol intake and tobacco smoking. Microsomal epoxide hydrolase (EPHX1) is a phase II detoxifying enzyme capable of tobacco-borne toxicant inactivation. We studied the role of the EPHX1 c.337T>C (p.Y113H) variant, whichleads to altered enzyme activity, in pancreatic diseases. METHODS: We genotyped 2391 patients by melting curve analysis. We enrolled 367 patients with pCA, 341 patients with alcoholic CP (aCP), 431 patients with idiopathic CP or hereditary pancreatitis, 192 patients with acute pancreatitis, and 679 controls of German descent. We replicated data in 77 patients with aCP and 304 controls from The Netherlands. RESULTS: In German patients with aCP, Y113 was more common than in controls (allele frequencies, 0.73 vs 0.68; risk ratio, 1.21 [95% confidence interval, 1.05-1.39]). However, we could not confirm this association in the Dutch population (allele frequencies, 0.62 vs 0.68, P=not significant). In total, Y113 frequency was 0.71 in aCP and 0.68 in controls (P = not significant). Allele frequencies did not differ in the other disease groups (acute pancreatitis, 0.69; idiopathic CP or hereditary pancreatitis, 0.68; pCA, 0.68; and control, 0.68). CONCLUSIONS: The EPHX1 Y113H variant is not associated with pancreatic diseases indicating that EPHX1 does not play a significant role in the initiation of pancreatic inflammation or cancer. PMID- 19287330 TI - Examination of external validity in randomized controlled trials for adjuvant treatment of pancreatic adenocarcinoma. AB - OBJECTIVES: Clinical trials are a prerequisite for evidence-based medicine. Trial representativity by internal and external validities is an assumption for a transfer into the real world. METHODS: A questionnaire addressing several forms of bias was established. Randomized controlled trials for adjuvant treatment of pancreatic adenocarcinoma with an evidence level of lower than 2 were selected and evaluated for internal and external validities focusing on selection bias. RESULTS: Four selected trials (European Study Group for Pancreatic Cancer 1 [ESPAC-1], Charite Onkologie Clinical Studies in GI Cancer 001 [CONKO 001], Radiation Therapy Oncology Group 97-04 [RTOG 97-04], and Adjuvant Chemoradioimmunotherapy of Pancreatic Carcinoma [CapRI]) proved to be evaluable. External validity was to a large extent given in the trials, whereby Radiation Therapy Oncology Group 97-04 included significantly more patients with better prognostic markers (negative for lymph nodes and T1/T2 tumors). Their inclusion and exclusion criteria are limited to the (unavoidable) restrictions for safety reasons and clinical restrictions. Comparison of participants with nonparticipants and with the target population via literature research proved to be preventive against selection and recruitment biases. A possible detection bias in the Charite Onkologie Clinical Studies in GI Cancer 001 trial cannot, however, be excluded. CONCLUSIONS: The analyzed trials have acceptable internal and external validities despite some minor criticisms. The protocols could be transferred into the clinical routine. It is recommended to open the inclusion criteria of trials wider and to implement the comparison with the target population. PMID- 19287331 TI - Sixty-one years of pancreatic cancer in Denmark from 1943 to 2003: a nationwide study. AB - OBJECTIVES: We report the incidence rates of pancreatic cancer in Denmark during 61 years of data registration, from 1943 to 2003. METHODS: On the basis of reports in the nationwide, population-based Danish Cancer Registry, we calculated age-standardized, period-specific incidence rates of pancreatic cancer. RESULTS: A total of 32,654 incident cases of pancreatic cancer were evaluated (male-female ratio, 1.4). The age-standardized incidence rate of pancreatic cancer increased steadily in the beginning of the study period from 3.75/100,000 person-years in 1943 to 1947 to the maximum of 9.96/100,000 person-years in 1968 to 1972 among men and from 2.95 in 1943 to 1947 to the maximum of 7.04 in 1978 to 1982 among women. The incidence rates declined between 1968 to 1972 and 1988 to 1992 for men and between 1978 to 1982 and 2003 for women. Most tumors were located in the exocrine pancreas, and most were adenocarcinomas. More than 40% were located in the head of the pancreas; 14% were localized, 21% were regionally spread, and 36% were metastatic at the time of diagnosis. During the period 1978 to 2003, the percentages of histologically or cytologically verified adenocarcinomas remained relatively steady, approximately 30%. CONCLUSIONS: The incidence rate of pancreatic cancer increased at the beginning of the 61 years of registration and declined slightly thereafter. PMID- 19287332 TI - Racial disparities in pancreatic cancer and radon exposure: a correlation study. AB - OBJECTIVES: To correlate radon exposure with the incidence of pancreatic cancer (PC) and to ascertain the influence of race in this correlation. METHODS: Age standardized incidence rates (SIRs) of PC from 1992 to 2002, segregated by race, were obtained from the Surveillance, Epidemiology, and End Results database. The mean radon levels for each county were obtained from the Environmental Protection Agency map, which assigns each county to 1 of 3 categories based on radon potential. Aggregate radon exposure was correlated to the aggregate SIRs of PC. RESULTS: The SIRs of PC in the United States ranged from 1.4 to 21.8/100,000 person-years. The highest rates for whites (19.6/100,000 person-years) and American Indians (594/100,000 person-years) were found in Guadalupe County, New Mexico; for African Americans (4845/100,000 person-years) in Worth County, Iowa; and for Asian Americans (3177/100,000 person-years) in Monroe County, Iowa. There was an insignificant correlation between radon exposure and overall incidence of PC (r = 0.042). A significant correlation existed between radon exposure and incidence of PC in African Americans, American Indians, and Asian Americans, but not in whites. CONCLUSIONS: Radon exposure may be a significant risk factor for PC in African Americans, American Indians, and Asian Americans. Testing and mitigating homes for indoor radon may decrease PC incidence in these groups. PMID- 19287333 TI - Diagnosis of small pancreatic cancer by endoscopic balloon-catheter spot pancreatography: an analysis of 29 patients. AB - OBJECTIVES: The diagnosis of small pancreatic cancer remains difficult. The present study describes the diagnostic value of endoscopic balloon-catheter spot pancreatography for small pancreatic cancer. METHODS: Since April 1984, balloon spot pancreatography has been used to detect small-sized pancreatic cancer in patients having possible symptoms or findings of obstructive pancreatitis. RESULTS: A resection was performed on 175 of 416 patients with conditions diagnosed as pancreatic cancer. Of the 175 patients, 23 (13%) had invasive carcinoma 2 cm or smaller based on histological measurements, 3 intraductal papillotubular adenocarcinoma, and 3 carcinoma in situ (CIS). Regarding invasive carcinoma, balloon pancreatography displayed duct abnormalities diagnosed as carcinoma in 20 of 22 patients, whereas carcinoma was suggested in 2. A definite diagnosis was obtained based on the findings of main duct stenosis or obstruction with marked stricture of the branch ducts (n = 18) and a filling defect in the main duct (n = 2). Moreover, this pancreatogram demonstrated an intraductal filling defect in 2 of 3 with intraductal carcinoma and dead twiglike findings in the branch ducts in 1 of 3 with CIS. CONCLUSIONS: Balloon spot pancreatography is an essential tool for the diagnosis of small ductal pancreatic cancer, and it also makes it possible to locate CIS lesions of the branch ducts. PMID- 19287334 TI - Comparison of endoscopic retrograde cholangiopancreatography, endoscopic ultrasonography, and fecal elastase 1 in chronic pancreatitis and clinical correlation. AB - OBJECTIVES: In this study, we prospectively compared the use of endoscopic retrograde cholangiopancreatography (ERCP), endoscopic ultrasonography (EUS), and fecal elastase 1 in patients with chronic pancreatitis and searched for correlation with symptoms, clinical findings, and elastase 1 levels. METHODS: Twenty-four consecutive patients (19 were male, and 5 were female) with chronic pancreatitis who had already undergone ERCP within the last 2 years and 19 healthy control subjects (10 were male, and 9 were female) are studied prospectively. Clinical and laboratory parameters of the patients were recorded, and all underwent EUS and fecal elastase 1 testing. Fecal elastase 1 was measured in healthy control subjects. RESULTS: The ERCP and EUS severity scores were 1 in 0 to 2 patients, 2 in 6 to 8 patients, and 3 in 18 to 14 patients. Sensitivity and specificity of fecal elastase for chronic pancreatitis were 75% and 100%, respectively. There was a negative correlation between disease duration and fecal elastase 1 levels. Patients with dyspepsia or those who use pancreatic enzyme preparations had significantly lower fecal elastase 1 levels than others. CONCLUSIONS: Endoscopic retrograde cholangiopancreatography and EUS are nearly equal in staging chronic pancreatitis. Fecal elastase 1 correlates well with these tests. Fecal elastase 1 also correlates well with some clinical symptoms such as dyspepsia and disease history. PMID- 19287335 TI - Comparison between mucinous cystic neoplasm and intraductal papillary mucinous neoplasm of the branch duct type of the pancreas with respect to expression of CD10 and cytokeratin 20. AB - OBJECTIVE: Mucinous cystic neoplasm (MCN) and intraductal papillary mucinous neoplasm of the branch duct type (IPMN-BD) differ in biological and clinical behaviors, but MCN is often misdiagnosed as IPMN-BD. The purpose of this study was to find useful markers for the differential diagnosis of MCN and IPMN-BD. METHODS: Immunohistochemically, the expression of the 2 types of mucin (MUC) 1 (MUC1/DF3 and MUC1/CORE), MUC2, MUC5AC, MUC6, human gastric mucin (HGM), caudal related homeobox transcription factor 2 (CDX2), CD10, cytokeratin (CK) 7, and CK20 was examined in 7 cases of MCN and 16 cases of IPMN-BD. RESULTS: Expression frequencies in MCN and IPMN-BD were 100% versus 44% for MUC1/DF3, 86% versus 31% for MUC1/CORE, 57% versus 19% for MUC2, 86% versus 100% for MUC5AC, 57% versus 88% for MUC6, 86% versus 100% for HGM, 57% versus 0% for CDX2, 71% versus 0% for CD10, 100% versus 69% for CK7, and 86% versus 6% for CK20. CONCLUSIONS: Mucin 1/DF3, MUC1/CORE, CDX2, CD10, and CK20 were expressed significantly more frequently in MCN than in IPMN-BD. In particular, CD10 and CK20 showed marked differences in immunohistochemical sensitivity and specificity between MCN and IPMN-BD. It is therefore proposed that CD10 and CK20 may be used for the differential diagnosis of MCN and IPMN-BD. PMID- 19287336 TI - Palmitate activates insulin signaling pathway in pancreatic rat islets. AB - OBJECTIVE: To investigate the action of palmitate on insulin receptor (IR) signaling pathway in rat pancreatic islets. The following proteins were studied: IR substrate-1 and -2 (IRS1 and IRS2), phosphatidylinositol 3-kinase, extracellular signal-regulated protein kinase-1 and -2 (ERK1/2), and signal transducer and activator of transcription 3 (STAT3). METHODS: Immunoblotting and immunoprecipitation assays were used to evaluate the phosphorylation states of IRS1 and IRS2 (tyrosine [Tyr]), ERK1/2 (threonine 202 [Thr202]/Tyr204), and STAT3 (serine [Ser727]). RESULTS: The exposure of rat pancreatic islets to 0.1-mmol/L palmitate for up to 30 minutes produced a significant increase of Tyr phosphorylation in IRS2 but not in IRS1. The association of phosphatidylinositol 3-kinase with IRS2 was also upregulated by palmitate. Exposure to 5.6-mmol/L glucose caused a gradual decrease in ERK1/2 (Thr202/Tyr204) and STAT3 (serine [Ser727]) phosphorylations after 30-minute incubation. The addition of palmitate (0.1 mmol/L), associated with 5.6-mmol/L glucose, abolished these latter effects of glucose after 15-minute incubation. CONCLUSIONS: Palmitate at physiological concentration associated with 5.6-mmol/L glucose activates IR signaling pathway in pancreatic beta cells. PMID- 19287337 TI - Down-regulation of hypoxia-inducible factor-1alpha by hyperbaric oxygen attenuates the severity of acute pancreatitis in rats. AB - OBJECTIVES: This study investigated the role of hypoxia-inducible factor 1alpha (HIF-1alpha) in acute pancreatitis (AP) and whether HIF-1alpha is involved in the therapeutic effects of hyperbaric oxygen (HBO) on AP. METHODS: Thirty Wistar rats with taurocholate-induced AP were randomly assigned to 3 groups (each group had 10 rats) receiving oxygen, HBO, or no therapeutic treatment 4 hours after induction. Ten healthy sham-operated rats also served as controls. The arterial oxygen saturation, PaO2, pH, lactate dehydrogenase in the arterial sera, and amylase and tumor necrosis factor alpha in the venous sera were measured 6 hours after induction. Pancreatic tissues were subjected to histopathologic analysis, immunohistochemical and Western-blotted analyses of HIF-1alpha and vascular endothelial growth factor, and measuring of myeloperoxidase activity. RESULTS: The HBO therapy attenuated the severity of acute pancreatitis; reduced histopathologic scores, dry weight-wet weight ratio of pancreatic tissues, and levels of amylase and lactate dehydrogenase; and elevated blood arterial oxygen saturation, PaO2, and pH values. The HBO therapy inhibited AP-induced up regulation of HIF-1alpha and its downstream effector vascular endothelial growth factor and the production of tumor necrosis factor alpha and myeloperoxidase activity. CONCLUSIONS: Hypoxia-inducible factor 1alpha plays a key role in the pathogenesis of AP, and the ability to down-regulate the expression of HIF-1alpha may partially explain the therapeutic effect of HBO on AP. PMID- 19287338 TI - Manipulation of gene expression by oxygen: a primer from bedside to bench. AB - For nearly 100 y, pediatricians have regularly used oxygen to treat neonatal and childhood diseases. During this time, it has become clear that oxygen is toxic and that overzealous use can lead to significant morbidity. As we have learned more about the appropriate clinical indications for oxygen therapy, studies at the bench have begun to elucidate the molecular mechanisms by which cells respond to hyperoxia. In this review, we discuss transcription factors whose activity is regulated by oxygen, including nuclear factor, erythroid 2-related factor 2 (Nrf2), activator protein 1 (AP-1), p53, nuclear factor kappaB (NF-kappaB), signal transducers and activators of transcription protein (STAT), and ccat/enhancer binding protein (CEBP). Special attention is paid to the mechanisms by which hyperoxia affects these transcription factors in the lung. Finally, we identify downstream targets of these transcription factors, with a focus on heme oxygenase-1. A better understanding of how oxygen affects various signaling pathways could lead to interventions aimed at preventing hyperoxic injury. PMID- 19287339 TI - Urine calcium/citrate ratio in children with hypercalciuric stones. AB - Hypercalciuria is a common cause for stone formation in children. The aim was to delineate the role of urinary citrate in hypercalciuric children for protection against calcium stone formation. We evaluated random urine calcium, citrate, and creatinine in 149 controls, 78 hypercalciuric nonstone formers, and 34 hypercalciuric children with stone. Urine citrate/creatinine was highest in hypercalciuric nonstone formers 899 +/- 351 compared with controls 711 +/- 328 and stone formers 595 +/- 289 (p < 0.01 vs. both). Calcium/creatinine ratio was similar in hypercalciuric stone and nonstone formers, but significantly higher than controls. Consequently, urine calcium/citrate ratio (mg/mg) increased from control 0.17 +/- 0.17 to 0.41 +/- 0.23 (p < 0.001) in hypercalciuric nonstone formers, and to 0.65 +/- 0.46 in stone formers (p < 0.001 compared with other groups). Area under receiver operating characteristic curve combined with multilevel risk analyses found calcium/citrate ratio of 0.326 to provide good discrimination between control and stone formers. We found 5th percentile for random urine citrate/creatinine ratio in school-aged children to be 176 mg/g, elevated urinary citrate excretion in hypercalciuric children to be protective against stone formation, and urine calcium/citrate ratio to be a good indicator for risk of stone formation. Whether intervention in hypercalciuric children to lower urine calcium/citrate <0.326 will provide protection against stone formation needs to be studied. PMID- 19287340 TI - Characterization of brain development in the ferret via MRI. AB - Animal models with complex cortical development are useful for improving our understanding of the wide spectrum of neurodevelopmental challenges facing human preterm infants. MRI techniques can define both cerebral injury and alterations in cerebral development with translation between animal models and the human infant. We hypothesized that the immature ferret would display a similar sequence of brain development [both gray (GM) and white matter (WM)] to that of the preterm human infant. We describe postnatal ferret neurodevelopment with conventional and diffusion MRI. The ferret is born lissencephalic with a thin cortical plate and relatively large ventricles. Cortical folding and WM maturation take place during the first month of life. From the mid-second through the third week of postnatal life, the ferret brain undergoes a similar, though less complex, pattern of maturational changes to those observed in the human brain during the second half of gestation. GM anisotropy decreases rapidly in the first 3 wks of life, followed by an upward surge of surface folding and WM anisotropy over the next 2 wks. PMID- 19287341 TI - Myocardial inflammation, cellular death, and viral detection in sudden infant death caused by SIDS, suffocation, or myocarditis. AB - The significance of minor myocardial inflammatory infiltrates and viral detection in SIDS is controversial. We retrospectively compared the demographic profiles, myocardial inflammation, cardiomyocyte necrosis, and myocardial virus detection in infants who died of SIDS in a safe sleep environment, accidental suffocation, or myocarditis. Formalin-fixed, paraffin-embedded myocardial sections were semiquantitatively assessed for CD3 lymphocytes and CD68 macrophages using immunohistochemistry and for cardiomyocyte cell death in H&E-stained sections. Enteroviruses and adenoviruses were searched for using PCR technology. The means of lymphocytes, macrophages, and necrotic cardiomyocytes were not statistically different in SIDS and suffocation cases. Enterovirus, not otherwise specified, was detected in one suffocation case and was the only virus detected in the three groups. Very mild myocardial lymphocyte and macrophage infiltration and scattered necrotic cardiomyocytes in SIDS are not pathologic, but may occur after the developing heart is exposed to environmental pathogens, including viruses. PMID- 19287342 TI - Effective lavage volume of diluted surfactant improves the outcome of meconium aspiration syndrome in newborn piglets. AB - Meconium aspiration syndrome (MAS) is one of the top causes of severe respiratory failure in neonates. This study was designed to investigate the effective volume of therapeutic bronchoalveolar lavage (BAL) with diluted surfactant in the treatment of MAS in newborn piglets. Human meconium was instilled in 24 piglets to induce MAS, and the piglets were randomly divided into four groups: 1) control, no lavage; 2) lavage-10, BAL with diluted surfactant (5 mg/mL, Survanta) 10 mL/kg in two aliquots; 3) lavage-20, 20 mL/kg in two aliquots; 4) lavage-30, 30 mL/kg in two aliquots. Cardiopulmonary parameters were monitored, and the lung tissue was histologically examined after experiments. The changes in oxygenation and lung compliance of lavage-20 and lavage-30 groups were significantly better than control and lavage-10 groups (p < 0.05), but there was no significant difference between lavage-20 and lavage-30 groups. The lung injury scores were significantly lower in the dependent site of lavage-20 and lavage-30 groups than the other two groups. In conclusion, using 20 mL/kg diluted surfactant in two aliquots to perform therapeutic BAL was as effective as 30 mL/kg in improving the pathophysiological outcomes in MAS and may warrant consideration clinically in treating MAS. PMID- 19287343 TI - Fetal growth restriction is associated with prioritization of umbilical blood flow to the left hepatic lobe at the expense of the right lobe. AB - Eighty to 85% of the venous perfusion to the fetal liver is from the umbilical vein, the rest from the portal vein. Umbilical venous flow to the liver is essential for intrauterine growth, and is impaired in placental insufficiency. We hypothesized that in growth-restricted fetuses portal blood flow compensates for insufficient umbilical blood flow to the liver. In 29 fetuses with fetal growth restriction (estimated fetal weight < or =5th percentile), we used ultrasound to measure blood flows in the umbilical vein, ductus venosus, left portal vein, and main portal stem. Compared with normal fetuses, both absolute and normalized total venous liver blood flows were reduced in growth-restricted fetuses, related to the degree of placental compromise and equally affecting both liver lobes. However, portal replaced umbilical flow to the right lobe, in a manner graded according to placental vascular resistance; in extreme cases, the right lobe received no umbilical perfusion. In fetal growth restriction, the liver suffers from venous hypoperfusion, and portal blood partially replaces umbilical flow to the right lobe; this will result in right liver lobe hypoxemia. This striking prioritization in nutrient delivery of left over right lobes suggests an adaptive response to poor placental perfusion that may have functional consequences. PMID- 19287344 TI - The human and mouse SLC25A29 mitochondrial transporters rescue the deficient ornithine metabolism in fibroblasts of patients with the hyperornithinemia hyperammonemia-homocitrullinuria (HHH) syndrome. AB - The hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome is a disorder of the urea cycle (UCD) and ornithine degradation pathway caused by mutations in the mitochondrial ornithine transporter (ORNT1). Unlike other UCDs, HHH syndrome is characterized by a less severe and variable phenotype that we believe may, in part, be due to genes with redundant function to ORNT1, such as the previously characterized ORNT2 gene. We reasoned that SLC25A29, a member of the same subfamily of mitochondrial carrier proteins as ORNT1 and ORNT2, might also have overlapping function with ORNT1. Here, we report that both the human and mouse SLC25A29, previously identified as mitochondrial carnitine/acyl carnitine transporter-like, when overexpressed transiently also rescues the impaired ornithine transport in cultured HHH fibroblasts. Moreover, we observed that, in the mouse, the Slc25a29 message is more significantly expressed in the CNS and cultured astrocytes when compared with the liver and kidney. These results suggest a potential physiologic role for the SLC25A29 transporter in the oxidation of fatty acids, ornithine degradation pathway, and possibly the urea cycle. Our results show that SLC25A29 is the third human mitochondrial ornithine transporter, designated as ORNT3, which may contribute to the milder and variable phenotype seen in patients with HHH syndrome. PMID- 19287345 TI - Histopathological changes of the heart after neonatal dexamethasone treatment: studies in 4-, 8-, and 50-week-old rats. AB - Dexamethasone (Dex), for prevention of chronic lung disease in preterm infants, showed potential negative long-term effects. Studies regarding long-term cardiovascular effects are lacking. We investigated possible histopathological myocardial changes after neonatal Dex in the young and adult rat heart. Rats were treated with Dex on d 1, 2, and 3 (0.5, 0.3, and 0.1 mg/kg) of life. Control-pups received saline. At 4, 8, and 50 wk after birth rats were killed and anatomic data collected. Heart tissue was stained with hematoxylin and eosin, Cadherin periodic acid schiff, and sirius red for cardiomyocyte morphometry and collagen determination. Presence of macrophages and mast cells was analyzed. Cardiomyocyte length of the Dex-treated rats was increased in all three age groups, whereas ventricular weight was reduced. Cardiomyocyte volumes were increased at 50 wk indicating cellular hypertrophy. Collagen content gradually increased with age and was 62% higher in Dex rats at 50 wk. Macrophage focus score and mast cell count were also higher. Neonatal Dex affects normal heart growth resulting in cellular hypertrophy and increased collagen deposition in the adult rat heart. Because previous studies in rats showed premature death, suggesting cardiac failure, cardiovascular follow-up of preterm infants treated with glucocorticoids should be considered. PMID- 19287346 TI - Exendin-4 normalizes islet vascularity in intrauterine growth restricted rats: potential role of VEGF. AB - Intrauterine growth restriction (IUGR) induced by uterine artery ligation in pregnant rats leads to low birth weight and early insulin secretory defects followed by the development of insulin resistance, decline in beta-cell mass, and diabetes in adulthood. Neonatal administration of Exendin-4 (Ex-4) prevents the deterioration of beta-cell mass and the onset of adult-onset diabetes. Our aim was to determine whether this effect occurs through preservation of islet vascularization. In 2 wk-old IUGR rats, endothelial-specific lectin staining revealed a 40% reduction in islet vascular density (p = 0.027), which was normalized by neonatal Ex-4. VEGF-A protein expression was reduced in IUGR islets compared with controls at postnatal d 1 (P). Neonatal Ex-4 normalized islet VEGF protein expression at P7. Neither IUGR nor Ex-4 administration to IUGR rats affected relative VEGF splice isoform RNA levels. Together, the reduced vascularity in IUGR islets before the deterioration of beta-cell mass, and the enhancement of VEGF expression and normalization of islet vascularity by neonatal Ex-4, suggest islet vascularity as an early determinant of beta-cell mass and as a potential therapeutic target for diabetes prevention. PMID- 19287347 TI - Intellectual, adaptive, and behavioral functioning in children with urea cycle disorders. AB - Inborn errors of urea synthesis lead to an accumulation of ammonia in blood and brain and result in high rates of mortality and neurodevelopmental disability. This study seeks to characterize the cognitive, adaptive, and emotional/behavioral functioning of children with urea cycle disorders (UCDs). These domains were measured through testing and parent questionnaires in 92 children with UCDs [33 neonatal onset (NO), 59 late onset (LO)]. Results indicate that children who present with NO have poorer outcome than those who present later in childhood. Approximately half of the children with NO performed in the range of intellectual disability (ID), including a substantial number ( approximately 30%) who were severely impaired. In comparison, only a quarter of the LO group was in the range of ID. There is also evidence that the UCD group has difficulties in aspects of emotional/behavioral and executive skills domains. In conclusion, children with UCDs present with a wide spectrum of cognitive outcomes. Children with NO disease have a much higher likelihood of having an ID, which becomes even more evident with increasing age. However, even children with LO UCDs demonstrate evidence of neurocognitive and behavioral impairment, particularly in aspects of attention and executive functioning. PMID- 19287348 TI - Plasma biomarkers in a mouse model of preterm labor. AB - Preterm labor (PTL) is frequently associated with inflammation. We hypothesized that biomarkers during pregnancy can identify pregnancies most at risk for development of PTL. An inflammation-induced mouse model of PTL was used. Surface enhanced laser desorption/ionization time-of-flight mass spectrometry was used to analyze and compare the plasma protein (PP) profile between CD-1 mice injected intrauterine with either lipopolysaccharide (LPS) or PBS on d 14.5 of gestation. The median differences of normalized PP peaks between the two groups were determined using the Mann-Whitney U test and the false discovery rate. In a second series of experiments, both groups of mice were injected with a lower dose of LPS. A total of 1665 peaks were detected. Thirty peaks were highly differentially expressed (p < 0.0001) between the groups. Two 11 kDa protein peaks were identified by MALDI-TOF/TOF-MS and confirmed to be mouse serum amyloid A (SAA) 1 and 2. Plasma SAA2 levels were increased in LPS-treated animals compared with controls and in LPS-treated animals that delivered preterm vs. those that delivered at term. SAA2 has the potential to be a plasma biomarker that can identify pregnancies at risk for development of PTL. PMID- 19287349 TI - Intrauterine growth restriction alters postnatal colonic barrier maturation in rats. AB - Intrauterine growth restriction (IUGR) is a leading cause of perinatal mortality and morbidity and increases the risk for necrotizing enterocolitis. We hypothesized that colonic barrier disruption could be responsible for intestinal frailty in infants and adults born with IUGR. Mucins and trefoil factor family 3 (TFF3) actively contribute to epithelium protection and healing. Our aim was to determine whether IUGR affects colonic mucosa maturation. IUGR was induced by dietary protein restriction in pregnant dams. Mucins and Tff3 expression and morphologic maturation of the colonic mucosa were followed during postnatal development of the offspring. Before weaning, mucin 2 and Tff3 protein levels were reduced in colonic mucosa of rats with IUGR compared with controls. After weaning, expression of mucin 2 (mRNA and protein) and mucin 4 (mRNA) were lower in colonic mucosa of rats with IUGR. At the same time, IUGR was associated with a reduction of crypt depth and a higher percentage of crypts in fission. We conclude that IUGR impairs mucus barrier development and is associated with long term alterations of mucin expression. The lack of an efficient colonic barrier induced by IUGR may predispose to colonic injury not only in neonatal life but also in later life. PMID- 19287350 TI - Cerebellar development in the preterm neonate: effect of supratentorial brain injury. AB - Cerebellar injury has been increasingly recognized as a complication of preterm birth, with decreased cerebellar volumes seen on follow-up neuroimaging. A cohort of 38 preterm newborns, including 14 with two scans, was studied with MRI, including single-shot fast spin-echo diffusion tensor imaging (DTI) sequence specifically to assess the posterior fossa. Early changes in the cerebellum [apparent diffusion coefficient (ADC) and fractional anisotropy (FA)] were assessed and correlated with supratentorial manifestations of injury [intraventricular hemorrhage (IVH) or white matter injury (WMI)]. ADC decreased and FA increased with increasing gestational age in both cerebellar gray and white matter. Severe IVH was associated with increased ADC in the middle cerebellar peduncles and hila of the cerebellar nuclei, decreased ADC in the cerebellar cortex, and decreased FA in all three regions. Changes with WMI were not consistent. Significant developmental changes in water diffusion were seen in cerebellar gray and white matter that were altered in patients with supratentorial IVH. DTI studies may provide an early indicator for cerebellar injury and abnormal cerebellar development in preterm neonates. PMID- 19287351 TI - Transmission of surfactant protein variants and haplotypes in children hospitalized with respiratory syncytial virus. AB - Severity of lung injury with respiratory syncytial virus (RSV) infection is variable and may be related to genetic variations. This preliminary report describes a prospective, family-based association study of children hospitalized secondary to RSV, aimed to determine whether intragenic and other haplotypes of surfactant proteins (SP)-A and SP-D are transmitted disproportionately from parents to offspring with RSV disease. Genomic DNA was genotyped for several SP-A and SP-D single nucleotide polymorphisms (SNPs). Transmission disequilibrium test analysis was used to determine transmission of variants and haplotypes from parents to affected offspring. Three hundred seventy-five individuals were studied, including 148 children with active RSV disease and one or both parents. The SP-A2 intragenic haplotype 1A was found to be protective (p = 0.013). The SP D SNP DA160_A may possibly be an "at-risk" marker (p = 0.0058). Additional two- and three-marker haplotypes were associated with severe RSV disease, with two being protective (DA11_T/DA160_G and DA160_G/SP-A2 1A/SP-A1 6A). We conclude that there may be associations between SP-A and SP-D and RSV disease. Further study is required to determine whether these variants can be used to target a high-risk patient population in clinical trials aimed at reducing either the symptoms of acute infection or long-term pulmonary sequelae. PMID- 19287352 TI - Implant complications, fusion, loss of lordosis, and outcome after anterior cervical plating with dynamic or rigid plates: two-year results of a multi centric, randomized, controlled study. AB - STUDY DESIGN: Prospective, controlled, randomized, multicenter study. OBJECTIVE: To analyze implant complications and speed. SUMMARY OF BACKGROUND DATA: Rigid plate designs, in which the screws are locked to the plate, are in common use and thought to provide more fixation than dynamic designs, in which the screws may glide when the graft is settling. The aim of the study is to analyze (1) implant complications, (2) speed of fusion, (3) loss of lordosis, and (4) clinical outcome in both types of plates. METHODS: One hundred thirty-two patients were included and assigned by randomization to one of the groups in which they received a routine anterior cervical discectomy and autograft fusion with either a dynamic plate (ABC, study group) or a rigid plate (CSLP, control group). At discharge, after 3 and 6 months and finally after 2 years, implant complications, segmental mobility, absence of radiolucencies, absence of bone sclerosis, evidence of bridging trabecular bone, loss of lordosis, Visual Analog Scale (VAS) and Neck Disability Score were recorded. All radiographic measurements were performed by an independent radiologist. RESULTS: There have been 4 patients with implant complications within the control group and no implant complications within the study group, P = 0.045. Mean segmental mobility before discharge for the study group was 1.7 mm, 1.4 mm after 3 months, 0.8 mm after 6 months, and 0.4 mm after 2 years. For the control group, these values were 1.0, 1.8, 1.6, and 0.5 mm. The difference at 6 months between both groups was significant (P = 0.024). Neither absence of radiolucencies, nor absence of sclerosis, nor evidence of bridging bone showed significant differences between the 2 groups through the postoperative follow-up (P > 0.05). The loss of segmental lordosis for the study group with respect to intraoperative radiograph was 1.3 degrees at discharge and 4.3 degrees after 2 years. For the control group, these values were 0.9 degrees , 0.7 degrees . The difference at 2 years was significant (P = 0.003). Clinical postoperative outcome (VAS and ODI) was not different between the 2 groups through the postoperative follow-up (P > 0.05). CONCLUSION: Dynamic cervical plate designs provide less implant complications (no patient) compared with rigid plate designs (4 patients). Speed of fusion was faster in the presence of a dynamic plate. However, loss of segmental lordosis is significantly higher if dynamic plates are used, which did not result in differences regarding clinical outcome between dynamic and constrained plates after 2 years. Thus, dynamic plates should be considered to be the preferred treatment option because of the lower risk for implant failure-related revision surgery. PMID- 19287353 TI - The preparation of Drosophila embryos for live-imaging using the hanging drop protocol. AB - Green fluorescent protein (GFP)-based timelapse live-imaging is a powerful technique for studying the genetic regulation of dynamic processes such as tissue morphogenesis, cell-cell adhesion, or cell death. Drosophila embryos expressing GFP are readily imaged using either stereoscopic or confocal microscopy. A goal of any live-imaging protocol is to minimize detrimental effects such as dehydration and hypoxia. Previous protocols for preparing Drosophila embryos for live-imaging analysis have involved placing dechorionated embryos in halocarbon oil and sandwiching them between a halocarbon gas-permeable membrane and a coverslip. The introduction of compression through mounting embryos in this manner represents an undesirable complication for any biomechanical-based analysis of morphogenesis. Our method, which we call the hanging drop protocol, results in excellent viability of embryos during live imaging and does not require that embryos be compressed. Briefly, the hanging drop protocol involves the placement of embryos in a drop of halocarbon oil that is suspended from a coverslip, which is, in turn, fixed in position over a humid chamber. In addition to providing gas exchange and preventing dehydration, this arrangement takes advantage of the buoyancy of embryos in halocarbon oil to prevent them from drifting out of position during timelapse acquisition. This video describes in detail how to collect and prepare Drosophila embryos for live imaging using the hanging drop protocol. This protocol is suitable for imaging dechorionated embryos using stereomicroscopy or any upright compound fluorescence microscope. PMID- 19287354 TI - American cutaneous leishmaniasis in children and adolescents from Northcentral Venezuela. AB - American Cutaneous Leishmaniasis (ACL) comprises a broad range of cutaneous manifestations caused by different Leishmania species which may produce severe and chronic sequelae in adults. However, it has been suggested that ACL may show different clinical and epidemiological features in children and adolescents that need to be further elucidated. We evaluated the epidemiological features of ACL in a cohort of pediatric patients from Northcentral Venezuela between years 1997 and 2005. Mean age of patients was 9 years old, with a mean clinical evolution of 3 months. Lesions were located mostly in extremities. Forty patients (93%) were positive by MST, 97.7% by IFAT and 48.8% by smear. MST values tended to be related to patients' age, higher values being recorded in older patients (p=0.153). PMID- 19287355 TI - Determination of malathion levels and the effect of malathion on the growth of Chrysomya megacephala (Fibricius) in malathion-exposed rat carcass. AB - This study was conducted to examine the effect of malathion on the development of Chrysomya megacephala. A total of 12 adult Sprague-Dawley rats was divided into 4 groups. Each animal in the 4 groups was given orally 0 (control), 10, 25 and 50ml/kg body weight of malathion, respectively. Chrysomya megacephala larvae were then allowed to grow on the liver of carcass. Larvae development was estimated by means of weight and length, time of adult emergence and survival rate. Results indicated that for the first 6 to 30 hours, larvae from control group developed more rapidly than larvae feeding on tissue containing malathion. However, the 3 doses of malathion did not exhibit significant impact on larvae length and weight. The time required for adult emergence was significantly greater for malathion-treated colony which was 10 days compared to 7 days in control colony. Control larvae of C. megacephala had higher survival rate compared to larvae exposed to the three different doses of malathion. Analysis of the tissues indicated that all rats and fly samples were positive for malathion. Malathion concentration was highest in liver. It was concluded that the presence of malathion altered the development rate of C. megacephala and thus disrupted normal postmortem interval estimation. PMID- 19287356 TI - Lack of insecticidal effect of mosquito coils containing either metofluthrin or esbiothrin on Anopheles gambiae sensu lato mosquitoes. AB - Use of mosquito coils for personal protection against malaria and mosquito nuisance is advocated under mosquito and malaria control programmes. We performed field studies of mosquito coils containing either metofluthrin or esbiothrin in experimental huts situated in Kamhororo village, Gokwe district, Zimbabwe. All tests were performed on 3-5 day old reared female Anopheles gambiae sensu lato mosquitoes. The burning times were 9hr 20min for mosquito coils containing metofluthrin and 8 hr for those containing esbiothrin and the results were significantly different (p = <0.001). The mean knock down rate for mosquito coils containing metofluthrin was 90% and that for esbiothrin was 73.3% and the results were significantly different (p = 0.00). Mosquito coils containing metofluthrin had a mean repellence of 92.7% as compared to 85.4% for esbiothrin and the results were not significantly different (p=0.27). The protection time as required by EPA (1999) was 6 hr for mosquito coils containing metofluthrin and 5 hr for those containing esbiothrin. The mean insecticidal effect of mosquito coils containing metofluthrin was 84% as compared to 83% for those containing esbiothrin and the results were not significantly different (p = 0.56). Both mosquito formulations could not be classified as having insecticidal effect since none of them met the 95% mortality rate criteria. PMID- 19287357 TI - A survey of caseous lymphadenitis in small ruminant farms from two districts in Perak, Malaysia -- Kinta and Hilir Perak. AB - A survey of Caseous Lymphadenitis (CLA), a bacterial infection in sheep and goats was conducted on small ruminant farms in two districts in Perak, namely Kinta and Hilir Perak. The objective of this survey is to determine the status of CLA infection in small ruminants. A total of 8 farms were screened, involving a total of 579 animals. Agar Gel Precipitation Test (AGPT) and Enzyme Linked Immuno Absorbent Assay (ELISA) were conducted on serum samples obtained from the animals. Results show that 8.5% of the animals had a positive reaction for AGPT test. It was found that 36 samples (17%) were found positive using both AGPT and ELISA methods, 9 samples (4%) were found positive only using AGPT method, 14 samples (6%)were found positive only using ELISA and 157 samples (73%) were found negative using both methods. Since there is no available data on the prevalence of the disease in the country, further epidemiological studies as well as reliable diagnostic detection methods need to be assessed for aiding in control and eradication programmes for this disease. PMID- 19287358 TI - Study of insect succession and rate of decomposition on a partially burned pig carcass in an oil palm plantation in Malaysia. AB - Insects found associated with corpse can be used as one of the indicators in estimating postmortem interval (PMI). The objective of this study was to compare the stages of decomposition and faunal succession between a partially burnt pig (Sus scrofa Linnaeus) and natural pig (as control). The burning simulated a real crime whereby the victim was burnt by murderer. Two young pigs weighed approximately 10 kg were used in this study. Both pigs died from pneumonia and immediately placed in an oil palm plantation near a pig farm in Tanjung Sepat, Selangor, Malaysia. One pig was partially burnt by 1-liter petrol while the other served as control. Both carcasses were visited twice per day for the first week and once thereafter. Adult flies and larvae on the carcasses were collected and later processed in a forensic entomology laboratory. Results showed that there was no significant difference between the rate of decomposition and sequence of faunal succession on both pig carcasses. Both carcasses were completely decomposed to remain stage after nine days. The species of flies visiting the pig carcasses consisted of blow flies (Chrysomya megacephala, Chrysomya rufifacies, Hemipyrellia ligurriens), flesh fly (Sarcophagidae.), muscid fly (Ophyra spinigera), soldier fly (Hermetia illucens), coffin fly (Phoridae) and scavenger fly (Sepsidae). The only difference noted was in the number of adult flies, whereby more flies were seen in the control carcass. Faunal succession on both pig carcasses was in the following sequence: Calliphoridae, Sarcophagidae, Muscidae, Phoridae and lastly Stratiomyidae. However, there was overlap in the appearance of members of these families. Blowflies continued to oviposit on both carcasses. Hence postmortem interval (PMI) can still be estimated from the partially burnt pig carcass. PMID- 19287359 TI - A study on the usefulness of Techlab Entamoeba histolytica II antigen detection ELISA in the diagnosis of amoebic liver abscess (ALA) at Hospital Universiti Sains Malaysia (HUSM), Kelantan, Malaysia. AB - Amoebic serodiagnosis at Hospital Universiti Sains Malaysia (HUSM), Kelantan employs an indirect haemagglutination assay (IHA) which detects anti-Entamoeba histolytica antibodies in patients' serum samples. In an amoebiasis endemic area such as Kelantan, interpretation of a positive IHA result can be problematic due to the high background antibody levels. The TechLab E. histolytica II ELISA is a commercial kit for detection of specific Gal/GalNAc lectin antigen in stool samples, and has been reported to be able to detect the antigen in serum samples from patients with amoebic liver abscess (ALA). Thus in this study we investigated the usefulness of TechLab E. histolytica II ELISA for diagnosis of ALA by comparing it with IHA. This is a cross sectional study involving 58 suspected ALA patients who were admitted to the surgical ward, HUSM, Kelantan. The diagnosis of ALA was established based on clinical symptoms and signs, ultrasound and/or CT scan results. The serum specimens obtained from the patients were tested with IHA (Dade Behring Diagnostics, Marburg, Germany) and TechLab E. histolytica II ELISA (Techlab, Blacksburg, Virginia, USA) according to the manufacturers' instructions. Of the 58 patients, 72.4% (42) were positive by IHA and only 8.6% (5) were positive by the TechLab E. histolytica II ELISA. Agreement between the IHA and ELISA was poor (kappa value 0.019, p=0.691). There was also no correlation between ELISA results and IHA antibody titers. The TechLab E. histolytica II ELISA was not sensitive in detecting amoebic antigen in samples from ALA patients. In addition the results of the test did not correlate with the IHA anti-E. histolytica antibody titres. Therefore, the TechLab E. histolytica II ELISA was found not to be useful for serological diagnosis of ALA at HUSM. PMID- 19287360 TI - Scanning electron micrographs of two species of Sturnophagoides (Acari: Astigmata: Pyroglyphidae) mites in Malaysia. AB - Scanning electron microscope (SEM) images of two dust mites, Sturnophagoides brasiliensis and Sturnophagoides halterophilus, are presented to provide an improved visualization of the taxonomic characters of these mites. Sturnophagoides halterophilus can be differentiated from S. brasiliensis by their expanded genu and femur of leg I. The differences in morphology of male and female S. brasiliensis are also discussed. PMID- 19287361 TI - Determination of the specificities of monoclonal and polyclonal antibodies to Neospora, Toxoplasma and Cryptosporidium by fluorescent antibody test (FAT). AB - Flourescent antibody test (FAT) was applied to determine the cross-reactivities of monoclonal (mAb), polyclonal (pAb) antibodies to Neospora, Toxoplasma and Cryptosporidium and antisera from cattle naturally infected with Neospora canium against antigens from a number of sources. Both mAb and pAb to Neospora reacted strongly (FAT titre up to 2560) with the homologous antigens and demonstrated weak titre (80) or no reaction with both Toxoplasma and Cryptosporidium antigens. Also mAb and pAb to Toxoplasma gondii reacted at titres of 80 - 640 with homologous antigens and at titres of 10-40 with N. caninum. No cross-reactions with either mAb or pAb antibodies to N. caninum and T. gondii were observed with Cryptosporidium parvum. The same results were observed with C. parvum mAb when tested with both N. caninum and T. gondii antigens. Sera from cattle naturally infected with N. caninum had titres ranging from 80- 640 with N. caninum antigens, and 10- 40 with T. gondii and C. parvum antigens. At low dilutions, the complete surfaces of Neospora and Toxoplasma parasites were fluorescent, while in higher dilutions only dotted fluorescence appeared on the apical complex. These results indicated the presence of cross-reactivity between Neospora and Toxoplasma but not with Cryptosporidium. Accordingly the recommended cut-off antibody titre for diagnosis of neosporosis is 80. PMID- 19287362 TI - Species composition of mosquito fauna in Ranau, Sabah, Malaysia. AB - The adult population and species composition of mosquitoes collected in Ranau, Sabah are described. A total of 5956 mosquitoes representing 8 genera and 41 species were collected using human landing catch, indoor and outdoor. Anopheles maculatus was the most common species (15.6%) followed by Culex quinquefasciatus (12.8%), Culex pseudovishnui (12.1%), Anopheles balabacensis (11.1%), Culex vishnui (9.7%), Aedes vexans (9.6%), Culex tritaeniorhyncus (6.6%), Anopheles donaldi (5.6%) and others in very small percentage. PMID- 19287363 TI - Intestinal parasitic infections in Srimum suburban area of Nakhon Ratchasima Province, Thailand. AB - A survey on intestinal parasitic infections and some risk factors of infection (social, economic and behavioral) was conducted in suburban area at Simum subdistrict, Mueang district, Nakhon Ratchasima Province during the period of April 3rd to April 11th, 2007. A total of 214 stool samples, from 85 males and 129 females were examined using simple direct smear, Kato's thick smear and modified Harada-Mori Filter Paper Strip culture technique. The results showed that the prevalence rate of protozoa infections by simple direct smear method was 17.3% (37 infected samples) in 11 males and 26 females and were from all age groups. Mixed infections (56.8%), showed higher results than single infection. The highest number of intestinal protozoan infections was Entamoeba histolytica (11.7%), followed by Blastocystis hominis (5.6%). Fifty four percent of protozoan infection was at a moderate to high level. Ninety five percent of infected cases consumed unboiled water. The prevalence rate of helminthic infections was 7% total by Kato's thick smear and Modified Harada-Mori Filter Paper Strip culture technique. Kato's Thick smear Technique showed the highest rate of Strongyloides stercolaris (2.8%), followed by hookworm infection (1.4%) Taenia spp. (0.9%), and Enterobius vermicularis (0.5%), respectively. All infections showed a light intensity, (< 200 epg), using Kato-Katz thick smear. Although Modified Harada Mori's culture Technique showed higher S. stercolaris (3.3%) and hookworm infection (2.3%) than Kato's thick smear in the area where there is low Soil transmitted helminthes, we recommend using Simple or Kato's thick smear to determine the prevalence of parasitic infection due to these techniques being able to detect various groups of parasites, and being inexpensive and less time consuming. PMID- 19287364 TI - Characterization of major allergens of royal jelly Apis mellifera. AB - Royal jelly is widely consumed in the community and has perceived benefits ranging from promoting growth in children and improvement of general health status to enhancement of longevity for the elderly. However, royal jelly consumption has been linked to contact dermatitis, acute asthma, anaphylaxis and death. High prevalence of positive skin tests to royal jelly have been reported among atopic populations in countries with a high rate of royal jelly consumption. The present study is aimed to identify the major allergens of royal jelly. Royal jelly extract was separated by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) and 2-dimensional electrophoresis (2-D). Immunoblotting of the SDS-PAGE and 2-D profiles were performed to identify the allergenic spots. Spots were then excised from the 2-D gel, digested with trypsin and analyzed by mass spectrometry. The SDS-PAGE of royal jelly extract revealed 18 bands between 10 to 167 kD. Western blot of the fractionated proteins detected 15 IgE-binding bands between 14 to 127 kD with seven major allergens of 32, 40, 42, 49, 55, 60 and 67 kD using serum from 53 subjects with royal jelly allergy. The 2-D gel fractionated the royal jelly proteins to more than 50 different protein spots. Out of these, 30 spots demonstrated specific IgE affinity to the sera tested. Eight spots of the major royal jelly allergens were selected for mass-spectrometry analysis. Digested tryptic peptides of the spots were compared to the amino acid sequence search in protein databases which identified the fragments of royal jelly homologus to major royal jelly protein 1 (MRJ1) and major royal jelly protein 2 (MRJ2). In conclusion, the major allergens of royal jelly are MRJ1 and MRJ2 in our patients' population. PMID- 19287365 TI - On the occurrence of Musca domestica L oviposition activity on pig carcass in peninsular Malaysia. AB - A forensic entomological study conducted in an oil palm plantation in Tanjung Sepat, Selangor, Malaysia on 3 August 2007 revealed that a housefly, Musca domestica Linnaeus oviposited its eggs on a freshly dead pig. This finding indicated that housefly might play an important role in forensic investigation in determining post-mortem interval (PMI), although it was not yet found in human corpses or any animal carrion. This preliminary paper presented a first record of Musca domestica eggs found on animal carcass in the country. PMID- 19287366 TI - A new record of Fannia prisca Stein, 1918 (Diptera:Fanniidae) from peninsular Malaysia. AB - Fannia prisca Stein, 1918 is newly recorded from peninsular Malaysia. This record is based on 4 male specimens from Mount Berembun, Brinchang, Cameron Highland, Pahang state, peninsular Malaysia. It is previously recorded from China, Mongolia, Korea, Japan, Taiwan, Bonin Island, Thailand and oriental region. The male of Fannia prisca can be differentiated from male Fannia scalaris by the following features: for F. prisca, mid-coxa without spine; mid-tibia normal or without stout triangular ventral projection; and hind tibia usually with 2 av, while F. scalaris has several stout hook-like spines on the anterior margin; mid tibia with stout triangular ventral projection and hind tibia usually with 3 av. Both F. prisca and F. scalaris can be differentiated from Fannia leucosticta by looking at its hind tibia, which only has 1 av. PMID- 19287367 TI - Seroprevalence of Toxoplasma gondii antibodies in pigs, goats, cattle, dogs and cats in peninsular Malaysia. AB - Antibodies to the protozoan parasite, Toxoplasma gondii were assayed in sera of 200 goats, 100 pigs, 126 cattle from various states of Malaysia, and 135 dogs and 55 cats around Ipoh region using an indirect fluorescence antibody test (IFAT, cut-off titer 1:200); antibodies were found in 35.5% of goats, 14.5% cats, 9.6% dogs, 7.9% local cattle and 4% yellow cattle but not in pigs. Results indicate that infection is most prevalent in goats. PMID- 19287368 TI - Occurrence of virulent genes among environmental isolates of Legionella pneumophila serogroup 1 strains from various parts of peninsular Malaysia. AB - Legionella pneumophila are intracellular pathogens, associated with human disease, attributed to the presence and absence of certain virulent genes. In this study, virulent gene loci (lvh and rtxA regions) associated with human disease were determined. Thirty-three cooling tower water isolates, isolated between 2004 to 2006, were analyzed for the presence of these genes by PCR method. Results showed that 19 of 33 (57.5%) of the L. pneumophila serogroup 1 isolates have both the genes. Six (18.2%) of the isolates have only the lvh gene and 2 (6.1%) of the isolates have only the rtxA gene. However, both genes were absent in 6 (18.2%) of the L. pneumophila isolates. The result of our study provides some insight into the presence of the disease causing L. pneumophila serogroup 1 in the environment. Molecular epidemiological studies will provide better understanding of the prevalence of the disease in Malaysia. PMID- 19287369 TI - A new record of Bengalia emarginata Malloch, 1927 (Diptera: Calliphoridae) from Malaysia. AB - During a forensic entomological study conducted at an oil palm plantation in Tanjung Sepat, Kuala Langat, Selangor, a Bengalia emarginata Malloch, 1927 (Diptera: Calliphoridae: Calliphorinae: Bengalini) was collected for the first time. Two adults were collected nearby the pig carcass by the first author and identified by the second. Prior to this finding, nine species of Bengalia were recorded from peninsular Malaysia or Borneo. Male of B. emarginata are different from Bengalia varicolor Fabricious by the following characters: Sternite 5 projection rounded with small identation and mid tibia double-fringed in ventral surface. PMID- 19287370 TI - First report of the signal fly, Scholastes sp. (Diptera:Platystomatidae) visiting animal carcasses in Malaysia. AB - Signal fly, Scholastes sp. (Diptera: Platystomatidae) was observed associated with animal carcasses in Malaysia. The first observation was on a monkey carcass, which was killed by using a handgun and immediately placed in a forested area in Gombak, Selangor while the second observation was on a pig that died of natural causes and whose carcass was placed in an oil palm plantation in Tanjung Sepat, Selangor. Both animal carcasses were visited by Scholastes sp. flies during the fresh decomposition period. However, the role Scholastes flies in the decomposition process remains unknown. In this paper, we report the occurrence of Scholastes sp. on animal carcasses in Malaysia for the first time. PMID- 19287371 TI - Phase II trial of Belagenpumatucel-L, a TGF-beta2 antisense gene modified allogeneic tumor vaccine in advanced non small cell lung cancer (NSCLC) patients. AB - In a previous dose escalation trial we demonstrated dose related survival correlation to Belagenpumatucel-L. In order to further evaluate safety and response at the previously defined optimal dose and schedule and to gain preliminary evidence on a hypothesis that the level of circulating tumor cells (CTCs) in blood may correlate with the overall survival of patients with stage IV NSCLC, we initiated a phase II trial. Patients received intradermal immunization of 2.5 x 10(7) transfected allogeneic tumor cells (Belagenpumatucel-L, supplied by NovaRx) 1 x every month for a total of 16 months. Circulating tumor cells (Veridex, Raritan, NJ) were measured every 4 weeks. Twenty-one advanced NSCLC patients were enrolled on this study. No significant toxic effect was observed. Overall survival was 562 days. The median survival was 660 days in patients having less than 2 CTCs at baseline compared to 150 days in patients with 2 or more CTCs (P=0.025). Phase II results of safety and response are consistent with prior experience following treatment with Belagenpumatucel-L and there is a suggestion that the number of circulating tumor cells at baseline appears to correlate with overall survival. A larger clinical trial is warranted to further explore this observation. PMID- 19287372 TI - SLC26A4 mutation spectrum associated with DFNB4 deafness and Pendred's syndrome in Pakistanis. AB - Pendred's syndrome (PDS) is an autosomal-recessive disorder characterized by sensorineural hearing loss and goiter. PDS is caused by mutations of the SLC26A4 gene encoding pendrin, a transmembrane exchanger of Cl(-), I(-) and HCO(3)(-), which is expressed in the thyroid and inner ear. SLC26A4 mutations can also be associated with non-syndromic deafness, DFNB4. The goal of our study was to define the identities and frequencies of SLC26A4 mutations in 563 large, consanguineous Pakistani families segregating severe-to-profound recessive deafness. Sequence analyses of SLC26A4 in 46 unreported families segregating deafness linked to DFNB4/PDS revealed 16 probable pathogenic variants, 8 of which are novel. The novel variants include three missense substitutions (p.R24L, p.G139V and p.V231M), two splice site mutations (c.304+2T>C and c.1341+3A>C), one frameshift (p.C565MfsX8) and two different genomic deletions affecting exons 1-2 and 11-18. Each of six pathogenic variants (p.V239D, p.Q446R, p.S90L, p.Y556C, p.R24L and p.K715N) was found in more than one family and haplotype analyses suggest that they are founder mutations. Combined with earlier reported data, SLC26A4 mutations were identified in 56 (7.2%; 95% CI: 5.6-9.2%) of 775 families. Therefore, SLC26A4 mutations are the most common known cause of genetic deafness in this population. As p.V239D (30%), p.S90L (18%) and p.Q446R (18%) account for approximately two-third of the mutant alleles of SLC26A4, hierarchical strategies for mutation detection would be feasible and cost-efficient genetic tests for DFNB4 deafness and PDS in Pakistanis. PMID- 19287373 TI - Broad diversity of neutralizing antibodies isolated from memory B cells in HIV infected individuals. AB - Antibodies to conserved epitopes on the human immunodeficiency virus (HIV) surface protein gp140 can protect against infection in non-human primates, and some infected individuals show high titres of broadly neutralizing immunoglobulin (Ig)G antibodies in their serum. However, little is known about the specificity and activity of these antibodies. To characterize the memory antibody responses to HIV, we cloned 502 antibodies from HIV envelope-binding memory B cells from six HIV-infected patients with broadly neutralizing antibodies and low to intermediate viral loads. We show that in these patients, the B-cell memory response to gp140 is composed of up to 50 independent clones expressing high affinity neutralizing antibodies to the gp120 variable loops, the CD4-binding site, the co-receptor-binding site, and to a new neutralizing epitope that is in the same region of gp120 as the CD4-binding site. Thus, the IgG memory B-cell compartment in the selected group of patients with broad serum neutralizing activity to HIV is comprised of multiple clonal responses with neutralizing activity directed against several epitopes on gp120. PMID- 19287374 TI - Gene regulatory logic of dopamine neuron differentiation. AB - Dopamine signalling regulates a variety of complex behaviours, and defects in dopamine neuron function or survival result in severe human pathologies, such as Parkinson's disease. The common denominator of all dopamine neurons is the expression of dopamine pathway genes, which code for a set of phylogenetically conserved proteins involved in dopamine synthesis and transport. Gene regulatory mechanisms that result in the direct activation of dopamine pathway genes and thereby ultimately determine the identity of dopamine neurons are poorly understood in all systems studied so far. Here we show that a simple cis regulatory element, the dopamine (DA) motif, controls the expression of all dopamine pathway genes in all dopaminergic cell types in Caenorhabditis elegans. The DA motif is activated by the ETS transcription factor AST-1. Loss of ast-1 results in the failure of all distinct dopaminergic neuronal subtypes to terminally differentiate. Ectopic expression of ast-1 is sufficient to activate the dopamine pathway in some cellular contexts. Vertebrate dopamine pathway genes also contain phylogenetically conserved DA motifs that can be activated by the mouse ETS transcription factor Etv1 (also known as ER81), and a specific class of dopamine neurons fails to differentiate in mice lacking Etv1. Moreover, ectopic Etv1 expression induces dopaminergic fate marker expression in neuronal primary cultures. Mouse Etv1 can also functionally substitute for ast-1 in C. elegans. Our studies reveal a simple and apparently conserved regulatory logic of dopamine neuron terminal differentiation and may provide new entry points into the diagnosis or therapy of conditions in which dopamine neurons are defective. PMID- 19287375 TI - Absence of nucleolar disruption after impairment of 40S ribosome biogenesis reveals an rpL11-translation-dependent mechanism of p53 induction. AB - Impaired ribosome biogenesis is attributed to nucleolar disruption and diffusion of a subset of 60S ribosomal proteins, particularly ribosomal protein (rp)L11, into the nucleoplasm, where they inhibit MDM2, leading to p53 induction and cell cycle arrest. Previously, we demonstrated that deletion of the 40S rpS6 gene in mouse liver prevents hepatocytes from re-entering the cell cycle after partial hepatectomy. Here, we show that this response leads to an increase in p53, which is recapitulated in culture by rpS6-siRNA treatment and rescued by the simultaneous depletion of p53. However, disruption of biogenesis of 40S ribosomes had no effect on nucleolar integrity, although p53 induction was mediated by rpL11, leading to the finding that the cell selectively upregulates the translation of mRNAs with a polypyrimidine tract at their 5'-transcriptional start site (5'-TOP mRNAs), including that encoding rpL11, on impairment of 40S ribosome biogenesis. Increased 5'-TOP mRNA translation takes place despite continued 60S ribosome biogenesis and a decrease in global translation. Thus, in proliferative human disorders involving hypomorphic mutations in 40S ribosomal proteins, specific targeting of rpL11 upregulation would spare other stress pathways that mediate the potential benefits of p53 induction. PMID- 19287376 TI - Characterization of the interface between normal and transformed epithelial cells. AB - In most cancers, transformation begins in a single cell in an epithelial cell sheet. However, it is not known what happens at the interface between non transformed (normal) and transformed cells once the initial transformation has occurred. Using Madin-Darby canine kidney (MDCK) epithelial cells that express constitutively active, oncogenic Ras (Ras(V12)) in a tetracycline-inducible system, we investigated the cellular processes arising at the interface between normal and transformed cells. We show that two independent phenomena occur in a non-cell-autonomous manner: when surrounded by normal cells, Ras(V12) cells are either apically extruded from the monolayer, or form dynamic basal protrusions and invade the basal matrix. Neither apical extrusion nor basal protrusion formation is observed when Ras(V12) cells are surrounded by other Ras(V12) cells. We show that Cdc42 and ROCK (also known as Rho kinase) have vital roles in these processes. We also demonstrate that E-cadherin knockdown in normal cells surrounding Ras(V12) cells reduces the frequency of apical extrusion, while promoting basal protrusion formation and invasion. These results indicate that Ras(V12)-transformed cells are able to recognize differences between normal and transformed cells, and consequently leave epithelial sheets either apically or basally, in a cell-context-dependent manner. PMID- 19287377 TI - p53-cofactor JMY is a multifunctional actin nucleation factor. AB - Many cellular structures are assembled from networks of actin filaments, and the architecture of these networks depends on the mechanism by which the filaments are formed. Several classes of proteins are known to assemble new filaments, including the Arp2/3 complex, which creates branched filament networks, and Spire, which creates unbranched filaments. We find that JMY, a vertebrate protein first identified as a transcriptional co-activator of p53, combines these two nucleating activities by both activating Arp2/3 and assembling filaments directly using a Spire-like mechanism. Increased levels of JMY expression enhance motility, whereas loss of JMY slows cell migration. When slowly migrating HL-60 cells are differentiated into highly motile neutrophil-like cells, JMY moves from the nucleus to the cytoplasm and is concentrated at the leading edge. Thus, JMY represents a new class of multifunctional actin assembly factor whose activity is regulated, at least in part, by sequestration in the nucleus. PMID- 19287378 TI - Myosin IIIa boosts elongation of stereocilia by transporting espin 1 to the plus ends of actin filaments. AB - Two proteins implicated in inherited deafness, myosin IIIa, a plus-end-directed motor, and espin, an actin-bundling protein containing the actin-monomer-binding motif WH2, have been shown to influence the length of mechanosensory stereocilia. Here we report that espin 1, an ankyrin repeat-containing isoform of espin, colocalizes with myosin IIIa at stereocilia tips and interacts with a unique conserved domain of myosin IIIa. We show that combined overexpression of these proteins causes greater elongation of stereocilia, compared with overexpression of either myosin IIIa alone or espin 1 alone. When these two proteins were co expressed in the fibroblast-like COS-7 cell line they induced a tenfold elongation of filopodia. This extraordinary filopodia elongation results from the transport of espin 1 to the plus ends of F-actin by myosin IIIa and depends on espin 1 WH2 activity. This study provides the basis for understanding the role of myosin IIIa and espin 1 in regulating stereocilia length, and presents a physiological example where myosins can boost elongation of actin protrusions by transporting actin regulatory factors to the plus ends of actin filaments. PMID- 19287379 TI - Store-operated cyclic AMP signalling mediated by STIM1. AB - Depletion of Ca(2+) from the endoplasmic reticulum (ER) results in activation of plasma membrane Ca(2+) entry channels. This 'store-operated' process requires translocation of a transmembrane ER Ca(2+) sensor protein, stromal interaction molecule 1 (STIM1), to sites closely apposed to Ca(2+) channels at the cell surface. However, it is not known whether a reduction in Ca(2+) stores is coupled to other signalling pathways by this mechanism. We found that lowering the concentration of free Ca(2+) in the ER, independently of the cytosolic Ca(2+) concentration, also led to recruitment of adenylyl cyclases. This resulted in enhanced cAMP accumulation and PKA activation, measured using FRET-based cAMP indicators. Translocation of STIM1 was required for efficient coupling of ER Ca(2+) depletion to adenylyl cyclase activity. We propose the existence of a pathway (store-operated cAMP signalling or SOcAMPS) in which the content of internal Ca(2+) stores is directly connected to cAMP signalling through a process that involves STIM1. PMID- 19287380 TI - Protein kinase DYRK2 is a scaffold that facilitates assembly of an E3 ligase. AB - Protein kinases have central functions in various cellular signal transduction pathways through their substrate phosphorylation. Here we show that a protein kinase, DYRK2, has unexpected role as a scaffold for an E3 ubiquitin ligase complex. DYRK2 associates with an E3 ligase complex containing EDD, DDB1 and VPRBP proteins (EDVP complex). Strikingly, DYRK2 serves as a scaffold for the EDVP complex, because small-interfering-RNA-mediated depletion of DYRK2 disrupts the formation of the EDD-DDB1-VPRBP complex. Although the kinase activity of DYRK2 is dispensable for its ability to mediate EDVP complex formation, it is required for the phosphorylation and subsequent degradation of its downstream substrate, katanin p60. Collectively, our results reveal a new type of E3 ubiquitin ligase complex in humans that depends on a protein kinase for complex formation as well as for the subsequent phosphorylation, ubiquitylation and degradation of their substrates. PMID- 19287381 TI - Ccbe1 is required for embryonic lymphangiogenesis and venous sprouting. AB - Lymphatic vessels have important roles in fluid homeostasis, fat absorption, inflammation and cancer metastasis and develop in a dynamic process (called lymphangiogenesis) involving budding, migration and proliferation of lymphangioblasts. Using a genetic screen in zebrafish we identify ccbe1 (collagen and calcium-binding EGF domain-1) as indispensible for embryonic lymphangiogenesis. Ccbe1 acts at the same stage of development as Vegfc and is required for lymphangioblast budding and angiogenic sprouting from venous endothelium. PMID- 19287382 TI - JAK2 haplotype is a major risk factor for the development of myeloproliferative neoplasms. AB - Chronic myeloproliferative neoplasms (MPNs) are a group of related conditions characterized by the overproduction of cells from one or more myeloid lineages. More than 95% of cases of polycythemia vera, and roughly half of essential thrombocythemia and primary myelofibrosis acquire a unique somatic 1849G>T JAK2 mutation (encoding V617F) that is believed to be a critical driver of excess proliferation. We report here that JAK2(V617F)-associated disease is strongly associated with a specific constitutional JAK2 haplotype, designated 46/1, in all three disease entities compared to healthy controls (polycythemia vera, n = 192, P = 2.9 x 10(-16); essential thrombocythemia, n = 78, P = 8.2 x 10(-9) and myelofibrosis, n = 41, P = 8.0 x 10(-5)). Furthermore, JAK2(V617F) specifically arises on the 46/1 allele in most cases. The 46/1 JAK2 haplotype thus predisposes to the development of JAK2(V617F)-associated MPNs (OR = 3.7; 95% CI = 3.1-4.3) and provides a model whereby a constitutional genetic factor is associated with an increased risk of acquiring a specific somatic mutation. PMID- 19287383 TI - Human mutation rate associated with DNA replication timing. AB - Eukaryotic DNA replication is highly stratified, with different genomic regions shown to replicate at characteristic times during S phase. Here we observe that mutation rate, as reflected in recent evolutionary divergence and human nucleotide diversity, is markedly increased in later-replicating regions of the human genome. All classes of substitutions are affected, suggesting a generalized mechanism involving replication time-dependent DNA damage. This correlation between mutation rate and regionally stratified replication timing may have substantial evolutionary implications. PMID- 19287384 TI - A germline JAK2 SNP is associated with predisposition to the development of JAK2(V617F)-positive myeloproliferative neoplasms. AB - Polycythemia vera, essential thrombocythemia and primary myelofibrosis are myeloproliferative neoplasms (MPN) characterized by multilineage clonal hematopoiesis. Given that the identical somatic activating mutation in the JAK2 tyrosine kinase gene (JAK2(V617F)) is observed in most individuals with polycythemia vera, essential thrombocythemia and primary myelofibrosis, there likely are additional genetic events that contribute to the pathogenesis of these phenotypically distinct disorders. Moreover, family members of individuals with MPN are at higher risk for the development of MPN, consistent with the existence of MPN predisposition loci. We hypothesized that germline variation contributes to MPN predisposition and phenotypic pleiotropy. Genome-wide analysis identified an allele in the JAK2 locus (rs10974944) that predisposes to the development of JAK2(V617F)-positive MPN, as well as three previously unknown MPN modifier loci. We found that JAK2(V617F) is preferentially acquired in cis with the predisposition allele. These data suggest that germline variation is an important contributor to MPN phenotype and predisposition. PMID- 19287385 TI - A common JAK2 haplotype confers susceptibility to myeloproliferative neoplasms. AB - Genome-wide association studies have identified a number of new disease susceptibility loci that represent haplotypes defined by numerous SNPs. SNPs within a disease-associated haplotype are thought to influence either the expression of genes or the sequence of the proteins they encode. In a series of investigations of the JAK2 gene in myeloproliferative neoplasms, we uncovered a new property of haplotypes that can explain their disease association. We observed a nonrandom distribution of the somatic JAK2(V617F) oncogenic mutation between two parental alleles of the JAK2 gene. We identified a haplotype that preferentially acquires JAK2(V617F) and confers susceptibility to myeloproliferative neoplasms. One interpretation of our results is that a certain combination of SNPs may render haplotypes differentially susceptible to somatic mutagenesis. Thus, disease susceptibility loci may harbor somatic mutations that have a role in disease pathogenesis. PMID- 19287386 TI - miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche. AB - The subventricular zone (SVZ) is the largest neurogenic niche in the adult mammalian brain. We found that the brain-enriched microRNA miR-124 is an important regulator of the temporal progression of adult neurogenesis in mice. Knockdown of endogenous miR-124 maintained purified SVZ stem cells as dividing precursors, whereas ectopic expression led to precocious and increased neuron formation. Furthermore, blocking miR-124 function during regeneration led to hyperplasias, followed by a delayed burst of neurogenesis. We identified the SRY box transcription factor Sox9 as being a physiological target of miR-124 at the transition from the transit amplifying cell to the neuroblast stage. Sox9 overexpression abolished neuronal differentiation, whereas Sox9 knockdown led to increased neuron formation. Thus miR-124-mediated repression of Sox9 is important for progression along the SVZ stem cell lineage to neurons. PMID- 19287387 TI - A dual leucine kinase-dependent axon self-destruction program promotes Wallerian degeneration. AB - Axon degeneration underlies many common neurological disorders, but the signaling pathways that orchestrate axon degeneration are unknown. We found that dual leucine kinase (DLK) [corrected to add (DLK) abbreviation] promoted degeneration of severed axons in Drosophila and mice, and that its target, c-Jun N-terminal kinase, promoted degeneration locally in axons as they committed to degenerate. This pathway also promoted degeneration after chemotherapy exposure and may be a component of a general axon self-destruction program. PMID- 19287388 TI - Proteoglycan interactions with Sonic Hedgehog specify mitogenic responses. AB - Sonic Hedgehog (Shh) has dual roles in vertebrate development, promoting progenitor cell proliferation and inducing tissue patterning. We found that the mitogenic and patterning functions of Shh can be uncoupled from one another. Using a genetic approach to selectively inhibit Shh-proteoglycan interactions in a mouse model, we found that binding of Shh to proteoglycans was required for proliferation of neural stem/precursor cells, but not for tissue patterning. Shh proteoglycan interactions regulated both spatial and temporal features of Shh signaling. Proteoglycans localized Shh to specialized mitogenic niches and also acted at the single-cell level to regulate the duration of Shh signaling, thereby promoting a gene expression program that is important for cell division. Because activation of the Shh pathway is a feature of diverse human cancers, selective stimulation of proliferation by Shh-proteoglycan interactions may also figure prominently in neoplastic growth. PMID- 19287389 TI - Traveling waves in developing cerebellar cortex mediated by asymmetrical Purkinje cell connectivity. AB - Correlated network activity is important in the development of many neural circuits. Purkinje cells are among the first neurons to populate the cerebellar cortex, where they sprout exuberant axon collaterals. We used multiple patch clamp recordings targeted with two-photon microscopy to characterize monosynaptic connections between the Purkinje cells of juvenile mice. We found that Purkinje cell axon collaterals projected asymmetrically in the sagittal plane, directed away from the lobule apex. On the basis of our anatomical and physiological characterization of this connection, we constructed a network model that robustly generated waves of activity that traveled along chains of connected Purkinje cells. Consistent with the model, we observed traveling waves of activity in Purkinje cells in sagittal slices from young mice that require GABA(A) receptor mediated transmission and intact Purkinje cell axon collaterals. These traveling waves are absent in adult mice, suggesting they have a developmental role in wiring the cerebellar cortical microcircuit. PMID- 19287390 TI - Endoplasmic reticulum stress in disorders of myelinating cells. AB - Myelinating cells, oligodendrocytes in the CNS and Schwann cells in the peripheral nervous system produce an enormous amount of plasma membrane during the myelination process, making them particularly susceptible to disruptions of the secretory pathway. Endoplasmic reticulum stress, initiated by the accumulation of unfolded or misfolded proteins, activates the unfolded protein response, which adapts cells to the stress. If this adaptive response is insufficient, the unfolded protein response activates an apoptotic program to eliminate the affected cells. Recent observations suggest that endoplasmic reticulum stress in myelinating cells is important in the pathogenesis of various disorders of myelin, including Charcot-Marie-Tooth disease, Pelizaeus-Merzbacher disease and Vanishing White Matter Disease, as well as in the most common myelin disorder, multiple sclerosis. A better understanding of endoplasmic reticulum stress in myelinating cells has laid the groundwork for the design of new therapeutic strategies for promoting myelinating cell survival in these disorders. PMID- 19287391 TI - Amyloid precursor protein secretases as therapeutic targets for traumatic brain injury. AB - Amyloid-beta (Abeta) peptides, found in Alzheimer's disease brain, accumulate rapidly after traumatic brain injury (TBI) in both humans and animals. Here we show that blocking either beta- or gamma-secretase, enzymes required for production of Abeta from amyloid precursor protein (APP), can ameliorate motor and cognitive deficits and reduce cell loss after experimental TBI in mice. Thus, APP secretases are promising targets for treatment of TBI. PMID- 19287392 TI - Deficiency of the intestinal enzyme acyl CoA:monoacylglycerol acyltransferase-2 protects mice from metabolic disorders induced by high-fat feeding. AB - Animals are remarkably efficient in absorbing dietary fat and assimilating this energy-dense nutrient into the white adipose tissue (WAT) for storage. Although this metabolic efficiency may confer an advantage in times of calorie deprivation, it contributes to obesity and associated metabolic disorders when dietary fat is abundant. Here we show that the intestinal lipid synthesis enzyme acyl CoA:monoacylglycerol acyltransferase-2 (MGAT2) has a crucial role in the assimilation of dietary fat and the accretion of body fat in mice. Mice lacking MGAT2 have a normal phenotype on a low-fat diet. However, on a high-fat diet, MGAT2-deficient mice are protected against developing obesity, glucose intolerance, hypercholesterolemia and fatty livers. Caloric intake is normal in MGAT2-deficient mice, and dietary fat is absorbed fully. However, entry of dietary fat into the circulation occurs at a reduced rate. This altered kinetics of fat absorption apparently results in more partitioning of dietary fat toward energy dissipation rather than toward storage in the WAT. Thus, our studies identify MGAT2 as a key determinant of energy metabolism in response to dietary fat and suggest that the inhibition of this enzyme may prove to be a useful strategy for treating obesity and other metabolic diseases associated with excessive fat intake. PMID- 19287393 TI - Cloning polymer single crystals through self-seeding. AB - In general, when a crystal is molten, all molecules forget about their mutual correlations and long-range order is lost. Thus, a regrown crystal does not inherit any features from an initially present crystal. Such is true for materials exhibiting a well-defined melting point. However, polymer crystallites have a wide range of melting temperatures, enabling paradoxical phenomena such as the coexistence of melting and crystallization. Here, we report a self-seeding technique that enables the generation of arrays of orientation-correlated polymer crystals of uniform size and shape ('clones') with their orientation inherited from an initial single crystal. Moreover, the number density and locations of these cloned crystals can to some extent be predetermined through the thermal history of the starting crystal. We attribute this unique behaviour of polymers to the coexistence of variable fold lengths in metastable crystalline lamellae, typical for ordering of complex chain-like molecules. PMID- 19287394 TI - Amplification-free Illumina sequencing-library preparation facilitates improved mapping and assembly of (G+C)-biased genomes. AB - Amplification artifacts introduced during library preparation for the Illumina Genome Analyzer increase the likelihood that an appreciable proportion of these sequences will be duplicates and cause an uneven distribution of read coverage across the targeted sequencing regions. As a consequence, these unfavorable features result in difficulties in genome assembly and variation analysis from the short reads, particularly when the sequences are from genomes with base compositions at the extremes of high or low G+C content. Here we present an amplification-free method of library preparation, in which the cluster amplification step, rather than the PCR, enriches for fully ligated template strands, reducing the incidence of duplicate sequences, improving read mapping and single nucleotide polymorphism calling and aiding de novo assembly. We illustrate this by generating and analyzing DNA sequences from extremely (G+C) poor (Plasmodium falciparum), (G+C)-neutral (Escherichia coli) and (G+C)-rich (Bordetella pertussis) genomes. PMID- 19287395 TI - TRF2 functions as a protein hub and regulates telomere maintenance by recognizing specific peptide motifs. AB - In mammalian cells, the telomeric repeat binding factor (TRF) homology (TRFH) domain-containing telomeric proteins TRF1 and TRF2 associate with a collection of molecules necessary for telomere maintenance and cell-cycle progression. However, the specificity and the mechanisms by which TRF2 communicates with different signaling pathways remain largely unknown. Using oriented peptide libraries, we demonstrate that the TRFH domain of human TRF2 recognizes [Y/F]XL peptides with the consensus motif YYHKYRLSPL. Disrupting the interactions between the TRF2 TRFH domain and its targets resulted in telomeric DNA-damage responses. Furthermore, our genome-wide target analysis revealed phosphatase nuclear targeting subunit (PNUTS) and microcephalin 1 (MCPH1) as previously unreported telomere-associated proteins that directly interact with TRF2 via the [Y/F]XL motif. PNUTS and MCPH1 can regulate telomere length and the telomeric DNA-damage response, respectively. Our findings indicate that an array of TRF2 molecules functions as a protein hub and regulates telomeres by recruiting different signaling molecules via a linear sequence code. PMID- 19287396 TI - Precursor-product discrimination by La protein during tRNA metabolism. AB - La proteins bind pre-tRNAs at their UUU-3'OH ends, facilitating their maturation. Although the mechanism by which La binds pre-tRNA 3' trailers is known, the function of the RNA binding beta-sheet surface of the RNA-recognition motif (RRM1) is unknown. How La dissociates from UUU-3'OH-containing trailers after 3' processing is also unknown. Here we show that La preferentially binds pre-tRNAs over processed tRNAs or 3' trailer products through coupled use of two sites: one on the La motif and another on the RRM1 beta-surface that binds elsewhere on tRNA. Two sites provide stable pre-tRNA binding, whereas the processed tRNA and 3' trailer are released from their single sites relatively fast. RRM1 loop-3 mutations decrease affinity for pre-tRNA and tRNA, but not for the UUU-3'OH trailer, and impair tRNA maturation in vivo. We propose that RRM1 functions in activities that are more complex than UUU-3'OH binding. Accordingly, the RRM1 mutations also impair an RNA chaperone activity of La. The results suggest how La distinguishes precursor from product RNAs, allowing it to recycle onto a new pre tRNA. PMID- 19287397 TI - The pathway of hepatitis C virus mRNA recruitment to the human ribosome. AB - Eukaryotic protein synthesis begins with mRNA positioning in the ribosomal decoding channel in a process typically controlled by translation-initiation factors. Some viruses use an internal ribosome entry site (IRES) in their mRNA to harness ribosomes independently of initiation factors. We show here that a ribosome conformational change that is induced upon hepatitis C viral IRES binding is necessary but not sufficient for correct mRNA positioning. Using directed hydroxyl radical probing to monitor the assembly of IRES-containing translation-initiation complexes, we have defined a crucial step in which mRNA is stabilized upon initiator tRNA binding. Unexpectedly, however, this stabilization occurs independently of the AUG codon, underscoring the importance of initiation factor-mediated interactions that influence the configuration of the decoding channel. These results reveal how an IRES RNA supplants some, but not all, of the functions normally carried out by protein factors during initiation of protein synthesis. PMID- 19287398 TI - A small molecule that directs differentiation of human ESCs into the pancreatic lineage. AB - Stepwise differentiation from embryonic stem cells (ESCs) to functional insulin secreting beta cells will identify key steps in beta-cell development and may yet prove useful for transplantation therapy for diabetics. An essential step in this schema is the generation of pancreatic progenitors--cells that express Pdx1 and produce all the cell types of the pancreas. High-content chemical screening identified a small molecule, (-)-indolactam V, that induces differentiation of a substantial number of Pdx1-expressing cells from human ESCs. The Pdx1-expressing cells express other pancreatic markers and contribute to endocrine, exocrine and duct cells, in vitro and in vivo. Further analyses showed that (-)-indolactam V works specifically at one stage of pancreatic development, inducing pancreatic progenitors from definitive endoderm. This study describes a chemical screening platform to investigate human ESC differentiation and demonstrates the generation of a cell population that is a key milepost on the path to making beta cells. PMID- 19287399 TI - Moving goal posts. PMID- 19287400 TI - Canine tongue trauma. PMID- 19287401 TI - Acronyms. PMID- 19287402 TI - Fabricated illness. PMID- 19287403 TI - Intuitive recording. PMID- 19287404 TI - Contrasting findings. PMID- 19287405 TI - A remarkable coincidence. PMID- 19287406 TI - Not an authority. PMID- 19287407 TI - Unsuitable prescribing. PMID- 19287408 TI - A hunger to buy. PMID- 19287409 TI - Young Dentist Conference explores 'worth'. PMID- 19287410 TI - Fluoride to be added to Southampton's water supply. PMID- 19287411 TI - Research finds no change to pregnancy outcomes. PMID- 19287419 TI - Dental professionalism: definitions and debate. AB - Professionalism has been identified as a core component of revalidation by the General Dental Council. However, analysis and debate over what it means to be a professional dentist is lacking in modern dentistry in the United Kingdom. The aim of this article is to open a debate on concepts of professionalism within dentistry, drawing on established thoughts in medicine and more limited material from the dental domain. The scope of discussion will extend to include definitions of professionalism, ethical issues within professionalism, professionalism in relation to revalidation and where all of these issues relate to dentistry perceived as a business. We can learn much from the medical community who have been driven to consider 'medical professionalism in a changing world', and in support of 'better patient care'. However, we can also contribute to the wider debate on professionalism by tackling the business angle, which has been largely ignored by our medical counterparts, and adding greater weight to the ethical implications of being a professional. PMID- 19287420 TI - A comparative needs assessment of the dental health of adults attending dental access centres and general dental practices in Halton & St Helens and Warrington PCTs 2007. AB - INTRODUCTION: Dental access centres (DACs) were introduced in England at the turn of the twenty-first century in response to a growing problem of access to NHS dental services. DACs were expected to offer NHS dental care primarily to those patients that were unwilling or unable to attend 'high street' dental practice. At the same time, the new NHS primary care dental contract in England, introduced in April 2006, has been associated in some areas with access difficulties, with routine dental patients having difficulty accessing NHS dental care. In light of these changes, have DACs become an alternative provider of NHS dental services to patients seeking routine dental care? METHOD: In summer 2007, a cross sectional dental epidemiological study was undertaken in Halton & St Helens PCT and Warrington PCT to compare the dental health and attitudes to dental visiting of adult patients attending DACs and neighbouring 'high street' dental practices. RESULTS: The results of the study showed that DAC patients: were younger and from a more disadvantaged background than patients attending 'high street' practices; had worse oral health than 'high street' dental patients; experienced more frequent episodes of dental pain than 'high street' dental patients and were more likely to be dentally anxious; had different attitudes to dental health than their 'high street' counterparts. CONCLUSIONS: The study suggests that the DACs in Halton, St Helens and Warrington are offering treatment to a different population of patients to that seen in neighbouring 'high street' practices and therefore the DACs are fulfilling the function expected of them locally. PMID- 19287421 TI - Facial atrophy following accidental subcutaneous extrusion of sodium hypochlorite. AB - Facial atrophy of the buccal fat pad following inadvertent subcutaneous extrusion of sodium hypochlorite is very rare. It is a complication which leaves the patient with a defect which is very difficult to treat. We present the case of a 46-year-old lady who presented with almost complete unilateral atrophy of the buccal fat pad. She has undergone multiple surgeries over the past two years. We highlight the precautions that must be taken to prevent such a complication and the difficulties in rehabilitating such a patient. PMID- 19287428 TI - Wenchuan earthquake: response of Chinese dental professionals. AB - On 12 May 2008, an earthquake with a magnitude of 8.0 on the Richter scale hit Wenchuan, China. In the aftermath of this natural disaster, Chinese dental professionals actively participated in the first emergency medical response team, definitive dental treatment, oral health services and education, and the recovery of local oral care infrastructure and resources. Learning from the experience and first-hand data of the Wenchuan earthquake, dental professionals can increase their awareness of the importance of collaborative emergency response health services in mass casualty events. Further research and emphasis is needed to encourage the participation of dental professionals in disaster preparation training and practice. PMID- 19287429 TI - Labial talon cusps: a South American archaeological case in the deciduous dentition and review of a rare trait. AB - Talon cusps are rarely reported developmental anomalies of the anterior dentition. They occur most commonly on the lingual aspect of the tooth but may also affect the labial aspect, and double (lingual and labial) talon cusps have also been reported. This study presents the first case of a labial talon cusp affecting the maxillary deciduous central incisor to be described in the literature. It was identified in a child skeleton from the archaeological site of Borgatta, north-west Argentina. The aetiology of talon cusps remains uncertain, with a combination of both genetic and environmental factors hypothesised to be involved. As a number of cases of labial talon cusps have now been published, this study presents a survey of cases in the literature to characterise the expression of this trait, and compares it with that of lingual talon cusps. The aim is to offer further insight into the relationship between lingual and labial talon cusps and their aetiologies. Differences between these traits in sex distribution, frequency of maxillary and mandibular cases, and the most commonly affected teeth are demonstrated. PMID- 19287447 TI - Sequencing the species pan-genome. PMID- 19287448 TI - Application of 'next-generation' sequencing technologies to microbial genetics. AB - New sequencing methods generate data that can allow the assembly of microbial genome sequences in days. With such revolutionary advances in technology come new challenges in methodologies and informatics. In this article, we review the capabilities of high-throughput sequencing technologies and discuss the many options for getting useful information from the data. PMID- 19287449 TI - Principles of c-di-GMP signalling in bacteria. AB - On the stage of bacterial signal transduction and regulation, bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) has long played the part of Sleeping Beauty. c-di-GMP was first described in 1987, but only recently was it recognized that the enzymes that 'make and break' it are not only ubiquitous in the bacterial world, but are found in many species in huge numbers. As a key player in the decision between the motile planktonic and sedentary biofilm-associated bacterial 'lifestyles', c-di-GMP binds to an unprecedented range of effector components and controls diverse targets, including transcription, the activities of enzymes and larger cellular structures. This Review focuses on emerging principles of c-di-GMP signalling using selected systems in different bacteria as examples. PMID- 19287452 TI - A Sino-British frontier workshop of cancer biology. PMID- 19287450 TI - Pathogenic neisseriae: surface modulation, pathogenesis and infection control. AB - Although renowned as a lethal pathogen, Neisseria meningitidis has adapted to be a commensal of the human nasopharynx. It shares extensive genetic and antigenic similarities with the urogenital pathogen Neisseria gonorrhoeae but displays a distinct lifestyle and niche preference. Together, they pose a considerable challenge for vaccine development as they modulate their surface structures with remarkable speed. Nonetheless, their host-cell attachment and invasion capacity is maintained, a property that could be exploited to combat tissue infiltration. With the primary focus on N. meningitidis, this Review examines the known mechanisms used by these pathogens for niche establishment and the challenges such mechanisms pose for infection control. PMID- 19287453 TI - Getting away from it all in Capri: the 2008 EMBO workshop on NF-kappaB. PMID- 19287454 TI - Distinct pools of cancer stem-like cells coexist within human glioblastomas and display different tumorigenicity and independent genomic evolution. AB - Glioblastomas (GBMs) contain transformed, self-maintaining, multipotent, tumour initiating cancer stem cells, whose identification has radically changed our perspective on the physiology of these tumours. Currently, it is unknown whether multiple types of transformed precursors, which display alternative sets of the complement of properties of true cancer stem cells, can be found in a GBM. If different subsets of such cancer stem-like cells (CSCs) do exist, they might represent distinct cell targets, with a differential therapeutic importance, also depending on their characteristics and lineage relationship. Here, we report the presence of two types of CSCs within different regions of the same human GBM. Cytogenetic and molecular analysis shows that the two types of CSCs bear quite diverse tumorigenic potential and distinct genetic anomalies, and, yet, derive from common ancestor cells. This provides critical information to unravel the development of CSCs and the key molecular/genetic components underpinning tumorigenicity in human GBMs. PMID- 19287455 TI - Tumor necrosis factor receptor-associated factor-1 enhances proinflammatory TNF receptor-2 signaling and modifies TNFR1-TNFR2 cooperation. AB - It has been shown that tumor necrosis factor receptor-2 (TNFR2) stimulation leads to degradation of TNF receptor associated factor-2 (TRAF2) and inhibition of TNFR1-induced activation of NFkappaB and JNK. Here, we show that TRAF1 inhibits TNFR2-induced proteasomal degradation of TRAF2 and relieves TNFR1-induced activation of NFkappaB from the inhibitory effect of TNFR2. TRAF1 co-recruited with TRAF2 to both TNF receptors. Despite lacking an amino-terminal RING/zinc finger domain, TRAF1 did not interfere with TNFR1-induced activation of JNK and NFkappaB. It is noted that physiological expression levels of TRAF1 enhanced NFkappaB activation and interleukin-8 (IL8) production induced by TNFR2. Thus, TRAF1 shifts the quality of integrated TNFR1-TNFR2 signaling from apoptosis induction to proinflammatory NFkappaB signaling. PMID- 19287456 TI - Cyclin D1b protein expression in breast cancer is independent of cyclin D1a and associated with poor disease outcome. AB - Aberrant expression of cyclin D1 protein is a common feature of breast cancer. However, the CCND1 gene encodes two gene products, cyclin D1a and cyclin D1b, which have discrete mechanisms of regulation and impact on cell behavior. A polymorphism at nucleotide 870 in the CCND1 gene, rs603965, influences the relative production of the encoded proteins and can impart increased risk for tumor development. Here, the impact of both the G/A870 polymorphism and cyclin D1b protein production on breast cancer risk, disease phenotype and patient outcome was analysed. In a large multiethnic case-control study, the G/A870 polymorphism conferred no significant risk for breast cancer overall or by stage or estrogen receptor (ER) status. However, the cyclin D1b protein was found to be upregulated in breast cancer, independent of cyclin D1a levels, and exhibited heterogeneous levels in breast cancer specimens. High cyclin D1a expression inversely correlated with the Ki67 proliferation marker and was not associated with clinical outcome. In contrast, elevated cyclin D1b expression was independently associated with adverse outcomes, including recurrence, distant metastasis and decreased survival. Interestingly, cyclin D1b was particularly associated with poor outcome in the context of ER-negative breast cancer. Thus, specific cyclin D1 isoforms are associated with discrete forms of breast cancer and high cyclin D1b protein levels hold prognostic potential. PMID- 19287457 TI - HHV8-positive, EBV-positive Hodgkin lymphoma-like large B-cell lymphoma and HHV8 positive intravascular large B-cell lymphoma. AB - Human herpesvirus type 8 (HHV8), also known as Kaposi's sarcoma-associated herpesvirus, is a human gamma herpesvirus that underlies the pathogenesis of Kaposi's sarcoma, primary effusion lymphoma and multicentric Castleman's disease. We recently encountered two cases of HHV8-positive large B-cell lymphoma with features not previously described. The first patient was a 61-year-old immunocompetent man with an enlarged cervical lymph node containing scattered large, bizarre cells in a reactive background of lymphocytes, plasma cells and scattered regressed follicles resembling those of hyaline-vascular Castleman's disease. The appearance suggested classical Hodgkin's lymphoma, but the large cells were negative for CD15, CD30, CD20 and CD3, and positive for MUM1/IRF4, EMA, HHV8, EBER and dim IgM lambda. The second patient was a 59-year-old HIV positive man who died after several weeks of fever, night sweats, anemia, thrombocytopenia, hepatosplenomegaly and multiorgan failure. At autopsy an intravascular large B-cell lymphoma that was positive for MUM1/IRF4, HHV8 and IgM lambda, and negative for CD20 and EBER involved multiple organs, including lung, heart, kidney, liver and spleen. On the basis of the histologic features in these two cases, the presence of HHV8 was unexpected. These cases expand the spectrum of lymphoproliferative disorders that can be associated with HHV8. PMID- 19287458 TI - Novel immunophenotypic features of marrow lymphoplasmacytic lymphoma and correlation with Waldenstrom's macroglobulinemia. AB - Lymphoplasmacytic lymphoma involving the bone marrow can be difficult to diagnose, and pathological features that predict the presence of associated Waldenstrom's macroglobulinemia have yet to be identified. To address these issues, marrow histology, immunohistochemistry, and flow cytometry were studied from 35 lymphoplasmacytic lymphoma cases that had comprehensive clinical assessment for Waldenstrom's macroglobulinemia. In all cases, the plasma cells were analyzed by a novel 6-color flow method. Both immunohistochemistry and flow cytometry were useful in identifying the lymphoid and plasmacytic disease components. In 19 cases, immunohistochemistry revealed an earlier unrecognized pattern of plasma cell infiltration in which they were physically separate from the lymphoid infiltrates. B-cell flow cytometry revealed monotypic cells in 96% of the cases. Approximately half of these were CD5 and/or CD23 positive, although none had features of chronic lymphocytic leukemia, and none of the B cells had flow cytometric features suggesting plasmacytic differentiation. In contrast, highly sensitive 6-color plasma cell flow cytometry revealed monotypic cells in 32 of the 35 cases; in 20 cases, the pattern of CD38 and CD138 coexpression detected was identical to that seen in plasma cell malignancies such as multiple myeloma. In 18 of these 20 lymphoplasmacytic lymphoma cases, these plasma cells were CD19 positive, distinguishing them from those of true plasma cell neoplasms, which are CD19 negative. It is interesting that the two lymphoplasmacytic lymphoma cases with CD19-negative plasma cells had an IgG isotype serum paraprotein. Apart from this, no other pathological correlates of the clinical or laboratory features of symptomatic Waldenstrom's macroglobulinemia were identified. PMID- 19287459 TI - GNAS1 mutations occur more commonly than previously thought in intramuscular myxoma. AB - Mutation detection plays an important role in diagnostic pathology, not only in providing a tissue diagnosis, but also in predicting response to antitumourigenic agents. However, mutation detection strategies are often hampered by masking of mutant alleles by wild-type sequences. Coamplification at lower denaturation temperature PCR (COLD-PCR) reportedly increases the proportion of rare variant sequences in a wild-type background by using PCR cycles in which the denaturation temperature is reduced to favour product formation with lower melt temperatures and heteroduplexes arising from minor variants. Intramuscular myxoma is a rare benign soft tissue neoplasm that occurs sporadically and less commonly in association with fibrous dysplasia (Mazabraud's syndrome). Fibrous dysplasia results from activating GNAS1 mutations, and the same mutations have been identified in small numbers of intramuscular myxoma. The aim of the study was primarily to establish whether COLD-PCR is more sensitive than conventional PCR; this was achieved by testing for GNAS1 mutations in intramuscular myxomas using the two methodologies. Mutations were detected in 8 of 28 (29%) cases of intramuscular myxomas using conventional PCR followed by mutation-specific restriction enzyme digestion (PCR-MSRED) whereas 17 of 28 (61%) mutations were detected using COLD-PCR/MSRED. Mutations were detected in two cases where a diagnosis of low-grade myxofibrosarcoma had been favoured over intramuscular myxoma. No mutations were detected in an additional 9 low-grade and 19 high-grade myxofibrosarcomas, and another 40 control samples. This study shows the power of COLD-PCR compared with conventional PCR in mutation detection, and shows that GNAS1 mutation detection increases diagnostic accuracy when distinguishing between intramuscular myxoma and low-grade myxofibrosarcoma. PMID- 19287460 TI - The surgical pathology of pulmonary infarcts: diagnostic confusion with granulomatous disease, vasculitis, and neoplasia. AB - Twenty-three cases of surgically resected pulmonary infarcts sent in consultation were reviewed to evaluate their morphology and to assess reasons for consultation. The morphology of these infarcts demonstrated that only a minority had the classical triangular shape at low magnification (26%) whereas the majority were either spherical (17%) or had a geographic pattern of necrosis (35%). The margin of the infarcted tissue often had a pseudogranulomatous appearance due to palisaded histiocytes, foam cells, or perpendicularly oriented proliferations of fibroblasts and myofibroblasts (74%) and occasional cholesterol and hemosiderin-laden giant cells. Basophilic granular karyorrhectic necrosis was seen focally (52%) as was vascular inflammation (56%) raising the differential diagnosis of Wegener's granulomatosis or infectious granulomas. These nonclassical features combined with a low incidence of clinical hemoptysis, chest pain and pleurisy, and a primary radiographic diagnosis of 'nodule r/o malignancy' highlight the need to consider thromboembolic pulmonary infarcts in the differential diagnosis of necrotic lung nodules with a histiocytic and fibroproliferative rim. PMID- 19287461 TI - Desmocollin-3: a new marker of squamous differentiation in undifferentiated large cell carcinoma of the lung. AB - Lung cancer classification in small-cell and non-small-cell types was recently challenged by data on the differential efficacy of new cytotoxic agents in specific histotypes. An accurate histotype definition has therefore gained interest in both preoperative and surgical materials, but is a hard task especially in undifferentiated large-cell tumors lacking morphological signs of squamous or glandular differentiation. The responsiveness of these latter subtypes to new drugs apparently more selective for adenocarcinomas or squamous carcinomas is not fully understood, also due to the heterogeneity of diagnostic criteria for this tumor entity. Current immunohistochemical markers are not fully specific and new molecules are to be explored. On the basis of gene expression profiling data, reporting a remarkable differential expression of desmocollin-3 (a protein localized in desmosomal junctions of stratified epithelial) between adeno- and squamous cancers, we immunostained 62 cases of resected undifferentiated large-cell lung carcinomas for desmocollin-3 (and for TTF-1, p63 and mucin stain), to test its ability to identify a (residual) squamous phenotype, if present. Desmocollin-3 was expressed in almost half of the undifferentiated large-cell cancers and was mutually exclusive with TTF-1 (positive in 39%; the remaining 18 % of cases was double negative). Special large cell carcinoma variants expressed desmocollin-3 in 6 of 6 basaloid, 7 of 12 clear cell types, again mutually exclusive with TTF-1 expression. None of seven sarcomatoid carcinomas reacted for either marker. In 31 cytological samples diagnosed as 'non-small-cell lung carcinoma', desmocollin-3 was again mutually exclusive with TTF-1 and stained all squamous carcinomas, 1 of 19 adenocarcinoma only, and 50% of large-cell carcinoma (all histologically confirmed). This combined morphophenotypic approach may represent a valid adjunct (for both surgical and cytological samples) in the selection of patients with lung cancer to medical treatments tailored according to different efficacy in different lung carcinomas of the squamous, adeno- and large-cell types. PMID- 19287462 TI - Aberrant survivin expression in endometrial hyperplasia: another mechanism of progestin resistance. AB - Up to 30% of failure rate in endometrial hyperplasia patients treated by progestin urges more detailed understanding of the mechanisms involved in progestin resistance. Survivin is a key regulator in the antiapoptotic network, and overexpression of survivin has been reported in endometrial hyperplasia and cancer. This study investigated the role of survivin in progestin resistance in endometrial hyperplasia. Pre- and post-treatment endometrial hyperplasia tissue samples from 23 women were examined for changes in survivin expression related to the administration of progestins. The impact of continuous or intermittent progestin treatment on survivin expression in Ishikawa cells was examined by the western blot. Survivin immunoreactivity was present in epithelial compartment of all pre-progestin-treated endometrial hyperplasia samples with mean nuclear indices 78 and cytoplasmic indices 114. In the 15 progestin responders, an average of 19.5-fold decrease of survivin expression was seen in epithelial nuclei (P<0.001) and 8-fold decrease in epithelial cytoplasm (P<0.001). In the eight non-responders, no significant changes in survivin expression were detected. With in vitro Ishikawa cells, survivin expression was effectively inhibited by either 72-h continuous treatment with 10 muM medroxyprogesterone acetate or 72 h after medroxyprogesterone acetate withdrawal. Our results indicated that dysregulation of survivin expression in hyperplastic endometrium may be part of the molecular mechanisms for progestin resistance. Intermittent, rather than continuous, progestin treatment may be more effective clinically for the treatment of endometrial hyperplasia. PMID- 19287463 TI - FGFR3 and TP53 mutation analysis in inverted urothelial papilloma: incidence and etiological considerations. AB - Urothelial papillomas and low-grade urothelial carcinomas have shown a high incidence of fibroblast growth factor receptor 3 (FGFR3) mutations and are associated with a favorable prognosis. The association of FGFR3 mutations with inverted papillomas is less known. We analyzed 20 cases of inverted papilloma in the urinary tract. Mutations of FGFR3 (exons 7, 10, and 15) and TP53 genes were evaluated by DNA sequencing in these cases. Point mutations of the FGFR3 gene were identified in 45% (9 of 20) of inverted papillomas with four cases exhibiting mutations at multiple exons. Seven cases had exon 7 mutations containing R248C, S249T, L259L, P260P, and V266M. Two cases had exon 10 and 15 mutations including A366D, H412H, E627D, D641N, and H643D; five cases had N653H. The most frequent mutation was identified at R248C. None of the inverted papillomas exhibited mutations in TP53. During a mean follow-up of 78 months, none had recurrence or developed urothelial carcinoma. These findings support the concept that low-grade and low-stage urothelial neoplasms arise in a background of molecular changes that are distinctly different from the molecular changes of high-grade and high-stage urothelial cancers. PMID- 19287464 TI - Novel endoscopic therapeutic modalities for superficial neoplasms arising in Barrett's esophagus: a primer for surgical pathologists. AB - This review introduces some of the novel endoscopic modalities used for the treatment of superficial neoplasms arising in the setting of Barrett's esophagus, namely endoscopic mucosal resection and photodynamic therapy. We describe the appropriate technical details for pathologists to know to effectively communicate with the gastroenterologists as well as the pitfalls in the evaluation of endoscopic mucosal resection specimens and post photodynamic therapy follow-up biopsies. PMID- 19287465 TI - Columnar cell lesions associated with breast calcifications on vacuum-assisted core biopsies: clinical, radiographic, and histological correlations. AB - Columnar cell lesions of the breast are increasingly recognized at mammography for their tendency to calcify. We studied 392 vacuum-assisted core biopsies performed solely for calcifications to evaluate the frequency of columnar cell lesions, their relationship with radiological risk, appearance of calcifications, and clinical data. Management and follow-up of columnar cell lesions without and with atypia (flat epithelial atypia) was analyzed. Cases with architectural atypia (cribriform spaces and/or micropapillae) were excluded from flat epithelial atypia. Calcifications were within the lumen of acini affected by columnar cell lesions in 137 out of 156 biopsies diagnosed with some columnar cell lesions. These represented 37% of vacuum-assisted core biopsies and 62% of low radiological risk (BI-RADS3) calcifications. High-risk (BI-RADS5) calcifications were never associated with columnar cell lesions. Age and menopausal status were comparable in columnar and in not-columnar cell lesions. Atypia was associated with long-term hormone replacement therapy in both lesions. Surgical biopsy was recommended for all cases with atypia. Flat epithelial atypia, as the only histological findings on vacuum-assisted core biopsies, was never associated with malignancy at surgery. In conclusion, we suggest that surgical excision is not mandatory when flat epithelial atypia is found as the most advanced lesion on vacuum-assisted core biopsy performed for low radiological risk calcifications, and that women should be advised of the possible hormone dependency of this entity. PMID- 19287466 TI - Specific expression of miR-17-5p and miR-127 in testicular and central nervous system diffuse large B-cell lymphoma. AB - Recent studies have shown that certain non-coding short RNAs, called miRNAs, play an important role in diffuse large B-cell lymphomas. Patients with diffuse large B-cell lymphoma have great diversity in both clinical characteristics, site of presentation and outcome. The aim of our study is to validate the differential expression in germinal center and non-germinal center diffuse large B-cell lymphoma,s and to study to the extent to which the primary site of differentiation is associated with the miRNA expression profile. We studied 50 cases of de novo diffuse large B-cell lymphoma for the expression of 15 miRNAs (miR-15a, miR-15b, miR-16, miR-17-3p, miR-17-5p, miR-18a, miR-19a, miR-19b, miR 20a, miR-21, miR-92, miR-127, miR-155, miR-181a and miR-221). Apart from 19 nodal cases without extranodal dissemination (stages I and II), we selected two groups with unambiguous stages I and II extranodal presentation; 9 cases of primary central nervous system, 11 cases of primary testicular and 11 cases of other primary extranodal diffuse large B-cell lymphomas. All cases were analyzed with qRT-PCR. In situ hybridization for the most differentially expressed miRNAs was performed to show miRNA expression in tumor cells, but not in background cells. MiR-21 and miR-19b showed the highest expression levels. No significant differences were seen between germinal center and non-germinal center diffuse large B-cell lymphomas in either the total or the nodal group for any of the 15 miRNAs. Two miRNAs showed significant differences in expression levels for diffuse large B-cell lymphoma subgroups according to the site of presentation. MiR-17-5p showed a significant higher expression level in the central nervous system compared with testicular and nodal diffuse large B-cell lymphomas (P<0.05). MiR-127 levels were significantly higher in testicular than in central nervous system and in nodal diffuse large B-cell lymphomas (P<0.05). We conclude that the location of diffuse large B-cell lymphoma is an important factor in determining the differential expression of miRNAs. PMID- 19287467 TI - New paradigms in the treatment of wet AMD: the impact of anti-VEGF therapy. AB - The incidence of age-related macular degeneration (AMD) in the United Kingdom is increasing with the ageing population. The wet form of this progressive and potentially blinding disease can develop very rapidly and lead to severe loss of central vision and reduction in quality of life, sometimes in just a matter of weeks. Recent advances in the treatment of wet AMD with the licensing of anti vascular endothelial growth factor therapies, coupled with current guidance from the Scottish Medicines Consortium and the National Institute of Health and Clinical Excellence have led to a subsequent increase in workload at AMD clinics due to the increased number of patients now eligible for treatment. In addition, the Royal College of Ophthalmologists recommend a 2-week diagnosis to treatment schedule due to the aggressive nature of the disease. The role of the retinal specialist is thus changing, and business management skills are becoming increasingly necessary to obtain the necessary resources to implement the guidance. Through prior experience and formal external evaluation of services at Frimley Park and Harrogate District Hospitals, a number of critical success factors have been developed for optimising treatment pathways in efficient wet AMD clinics. PMID- 19287480 TI - Wiring and plumbing in the brain. PMID- 19287481 TI - Receptor-induced dilatation in the systemic and intrarenal adaptation to pregnancy in rats. AB - Normal pregnancy is associated with systemic and intrarenal vasodilatation resulting in an increased glomerular filtration rate. This adaptive response occurs in spite of elevated circulating levels of angiotensin II (Ang II). In the present study, we evaluated the potential mechanisms responsible for this adaptation. The reactivity of the mesangial cells (MCs) cultured from 14-day pregnant rats to Ang II was measured through changes in the intracellular calcium concentration ([Cai]). The expression levels of inducible nitric oxide synthase (iNOS), the Ang II-induced vasodilatation receptor AT2, and the relaxin (LGR7) receptor were evaluated in cultured MCs and in the aorta, renal artery and kidney cortex by real time-PCR. The intrarenal distribution of LGR7 was further analyzed by immunohistochemistry. The MCs displayed a relative insensitivity to Ang II, which was paralleled by an impressive increase in the expression level of iNOS, AT2 and LGR7. These results suggest that the MCs also adapt to the pregnancy, thereby contributing to the maintenance of the glomerular surface area even in the presence of high levels of Ang II. The mRNA expression levels of AT2 and LGR7 also increased in the aorta, renal artery and kidney of the pregnant animals, whereas the expression of the AT1 did not significantly change. This further suggests a role of these vasodilatation-induced receptors in the systemic and intrarenal adaptation during pregnancy. LGR7 was localized in the glomeruli and on the apical membrane of the tubular cells, with stronger labeling in the kidneys of pregnant rats. These results suggest a role of iNOS, AT2, and LGR7 in the systemic vasodilatation and intrarenal adaptation to pregnancy and also suggest a pivotal role for relaxin in the tubular function during gestation. PMID- 19287482 TI - Probing cellular dynamics with a chemical signal generator. AB - Observations of material and cellular systems in response to time-varying chemical stimuli can aid the analysis of dynamic processes. We describe a microfluidic "chemical signal generator," a technique to apply continuously varying chemical concentration waveforms to arbitrary locations in a microfluidic channel through feedback control of the interface between parallel laminar (co flowing) streams. As the flow rates of the streams are adjusted, the channel walls are exposed to a chemical environment that shifts between the individual streams. This approach can be used to probe the dynamic behavior of objects or substances adherent to the interior of the channel. To demonstrate the technique, we exposed live fibroblast cells to ionomycin, a membrane-permeable calcium ionophore, while assaying cytosolic calcium concentration. Through the manipulation of the laminar flow interface, we exposed the cells' endogenous calcium handling machinery to spatially-contained discrete and oscillatory intracellular disturbances, which were observed to elicit a regulatory response. The spatiotemporal precision of the generated signals opens avenues to previously unapproachable areas for potential investigation of cell signaling and material behavior. PMID- 19287483 TI - Evidence that plasmid-borne botulinum neurotoxin type B genes are widespread among Clostridium botulinum serotype B strains. AB - BACKGROUND: Plasmids that encode certain subtypes of the botulinum neurotoxin type B have recently been detected in some Clostridium botulinum strains. The objective of the present study was to investigate the frequency with which plasmid carriage of the botulinum neurotoxin type B gene (bont/B) occurs in strains of C. botulinum type B, Ab, and A(B), and whether plasmid carriage is bont/B subtype-related. METHODOLOGY/PRINCIPAL FINDINGS: PCR-Restriction fragment length polymorphism was employed to identify subtypes of the bont/B gene. Pulsed field gel electrophoresis and Southern blot hybridization with specific probes were performed to analyze the genomic location of the bont/B subtype genes. All five known bont/B subtype genes were detected among the strains; the most frequently detected subtype genes were bont/B1 and /B2. Surprisingly, the bont/B subtype gene was shown to be plasmid-borne in >50% of the total strains. The same bont/B subtype gene was associated with the chromosome in some strains, whereas it was associated with a plasmid in others. All five known bont/B subtype genes were in some cases found to reside on plasmids, though with varying frequency (e.g., most of the bont/B1 subtype genes were located on plasmids, whereas all but one of the bont/B2 subtypes were chromosomally-located). Three bivalent isolates carried both bont/A and /B genes on the same plasmid. The plasmids carrying the bont gene were five different sizes, ranging from approximately 55 kb to approximately 245 kb. CONCLUSIONS/SIGNIFICANCE: The unexpected finding of the widespread distribution of plasmids harboring the bont/B gene among C. botulinum serotype B strains provides a chance to examine their contribution to the dissemination of the bont genes among heterogeneous clostridia, with potential implications on issues related to pathogenesis and food safety. PMID- 19287484 TI - Clique-finding for heterogeneity and multidimensionality in biomarker epidemiology research: the CHAMBER algorithm. AB - BACKGROUND: Commonly-occurring disease etiology may involve complex combinations of genes and exposures resulting in etiologic heterogeneity. We present a computational algorithm that employs clique-finding for heterogeneity and multidimensionality in biomedical and epidemiological research (the "CHAMBER" algorithm). METHODOLOGY/PRINCIPAL FINDINGS: This algorithm uses graph-building to (1) identify genetic variants that influence disease risk and (2) predict individuals at risk for disease based on inherited genotype. We use a set covering algorithm to identify optimal cliques and a Boolean function that identifies etiologically heterogeneous groups of individuals. We evaluated this approach using simulated case-control genotype-disease associations involving two and four-gene patterns. The CHAMBER algorithm correctly identified these simulated etiologies. We also used two population-based case-control studies of breast and endometrial cancer in African American and Caucasian women considering data on genotypes involved in steroid hormone metabolism. We identified novel patterns in both cancer sites that involved genes that sulfate or glucuronidate estrogens or catecholestrogens. These associations were consistent with the hypothesized biological functions of these genes. We also identified cliques representing the joint effect of multiple candidate genes in all groups, suggesting the existence of biologically plausible combinations of hormone metabolism genes in both breast and endometrial cancer in both races. CONCLUSIONS: The CHAMBER algorithm may have utility in exploring the multifactorial etiology and etiologic heterogeneity in complex disease. PMID- 19287485 TI - The role of p300 histone acetyltransferase in UV-induced histone modifications and MMP-1 gene transcription. AB - Matrix metalloproteinase (MMP)-1 promotes ultraviolet (UV)-triggered long-term detrimental effects such as cancer formation and premature skin aging. Although histone modifications may play a crucial role in the transcriptional regulation of MMP-1, the relationship between UV-induced histone modification and MMP-1 expression is not completely understood. Here, we identify regulators of histone acetylation that may link UV-mediated DNA damage and MMP-1 induction by UV in cultured human dermal fibroblasts (HDFs) in vitro. UV irradiation of HDFs induced MMP-1 expression and increased the level of phosphorylation of H2AX (gamma-H2AX), p53 and the acetylation of histone H3 (acetyl-H3). Total histone deacetylase (HDAC) enzymatic activity was decreased by UV irradiation, while histone acetyltransferase (HAT) activity was increased. Suppression of p300 histone acetyltransferase (p300HAT) activity by the p300HAT inhibitor anacardic acid (AA) or by down-regulation of p300 by siRNA prevented UV-induced MMP-1 expression and inhibited UV-enhanced gamma-H2AX, p53 level, and acetyl-H3. Using chromatin immunoprecipitation assays, we observed that gamma-H2AX, p53, acetyl-H3, p300 and c-Jun were consistently recruited by UV to a distinct region (-2067/-1768) adjacent to the p300 binding site (-1858/-1845) in the MMP-1 promoter. In addition, these recruitments of gamma-H2AX, p53, acetyl-H3, p300 and c-Jun to the p300-2 site were significantly abrogated by post-treatment with AA. Furthermore, overexpression of p300 increased the basal and UV-induced MMP-1 promoter activity. Our results suggest that p300HAT plays a critical role in the transcriptional regulation of MMP-1 by UV. PMID- 19287486 TI - Learning of arbitrary association between visual and auditory novel stimuli in adults: the "bond effect" of haptic exploration. AB - BACKGROUND: It is well-known that human beings are able to associate stimuli (novel or not) perceived in their environment. For example, this ability is used by children in reading acquisition when arbitrary associations between visual and auditory stimuli must be learned. The studies tend to consider it as an "implicit" process triggered by the learning of letter/sound correspondences. The study described in this paper examined whether the addition of the visuo-haptic exploration would help adults to learn more effectively the arbitrary association between visual and auditory novel stimuli. METHODOLOGY/PRINCIPAL FINDINGS: Adults were asked to learn 15 new arbitrary associations between visual stimuli and their corresponding sounds using two learning methods which differed according to the perceptual modalities involved in the exploration of the visual stimuli. Adults used their visual modality in the "classic" learning method and both their visual and haptic modalities in the "multisensory" learning one. After both learning methods, participants showed a similar above-chance ability to recognize the visual and auditory stimuli and the audio-visual associations. However, the ability to recognize the visual-auditory associations was better after the multisensory method than after the classic one. CONCLUSION/SIGNIFICANCE: This study revealed that adults learned more efficiently the arbitrary association between visual and auditory novel stimuli when the visual stimuli were explored with both vision and touch. The results are discussed from the perspective of how they relate to the functional differences of the manual haptic modality and the hypothesis of a "haptic bond" between visual and auditory stimuli. PMID- 19287487 TI - Stepwise development of hematopoietic stem cells from embryonic stem cells. AB - The cellular ontogeny of hematopoietic stem cells (HSCs) remains poorly understood because their isolation from and their identification in early developing small embryos are difficult. We attempted to dissect early developmental stages of HSCs using an in vitro mouse embryonic stem cell (ESC) differentiation system combined with inducible HOXB4 expression. Here we report the identification of pre-HSCs and an embryonic type of HSCs (embryonic HSCs) as intermediate cells between ESCs and HSCs. Both pre-HSCs and embryonic HSCs were isolated by their c-Kit(+)CD41(+)CD45(-) phenotype. Pre-HSCs did not engraft in irradiated adult mice. After co-culture with OP9 stromal cells and conditional expression of HOXB4, pre-HSCs gave rise to embryonic HSCs capable of engraftment and long-term reconstitution in irradiated adult mice. Blast colony assays revealed that most hemangioblast activity was detected apart from the pre-HSC population, implying the early divergence of pre-HSCs from hemangioblasts. Gene expression profiling suggests that a particular set of transcripts closely associated with adult HSCs is involved in the transition of pre-HSC to embryonic HSCs. We propose an HSC developmental model in which pre-HSCs and embryonic HSCs sequentially give rise to adult types of HSCs in a stepwise manner. PMID- 19287488 TI - Interaction of the coronavirus infectious bronchitis virus membrane protein with beta-actin and its implication in virion assembly and budding. AB - Coronavirus M protein is an essential component of virion and plays pivotal roles in virion assembly, budding and maturation. The M protein is integrated into the viral envelope with three transmembrane domains flanked by a short amino-terminal ectodomain and a large carboxy-terminal endodomain. In this study, we showed co purification of the M protein from coronavirus infectious bronchitis virus (IBV) with actin. To understand the cellular factors that may be involved in virion assembly, budding and maturation processes, IBV M was used as the bait in a yeast two-hybrid screen, resulting in the identification of beta-actin as a potentially interacting partner. This interaction was subsequently confirmed by coimmunoprecipitation and immunofluorescence microscopy in mammalian cells, and mutation of amino acids A159 and K160 on the M protein abolished the interaction. Introduction of the A159-K160 mutation into an infectious IBV clone system blocks the infectivity of the clone, although viral RNA replication and subgenomic mRNA transcription were actively detected. Disruption of actin filaments with cell permeable agent cytochalasin D at early stages of the infection cycle led to the detection of viral protein synthesis in infected cells but not release of virus particles to the cultured media. However, the same treatment at late stages of the infection cycle did not affect the release of virus particles to the media, suggesting that disruption of the actin filaments might block virion assembly and budding, but not release of the virus particles. This study reveals an essential function of actin in the replication cycle of coronavirus. PMID- 19287489 TI - HIV gp120 induces, NF-kappaB dependent, HIV replication that requires procaspase 8. AB - BACKGROUND: HIV envelope glycoprotein gp120 causes cellular activation resulting in anergy, apoptosis, proinflammatory cytokine production, and through an unknown mechanism, enhanced HIV replication. METHODOLOGY/PRINCIPAL FINDINGS: We describe that the signals which promote apoptosis are also responsible for the enhanced HIV replication. Specifically, we demonstrate that the caspase 8 cleavage fragment Caspase8p43, activates p50/p65 Nuclear Factor kappaB (NF-kappaB), in a manner which is inhibited by dominant negative IkappaBalpha. This caspase 8 dependent NF-kappaB activation occurs following stimulation with gp120, TNF, or CD3/CD28 crosslinking, but these treatments do not activate NF-kappaB in cells deficient in caspase 8. The Casp8p43 cleavage fragment also transactivates the HIV LTR through NF-kappaB, and the absence of caspase 8 following HIV infection greatly inhibits HIV replication. CONCLUSION/SIGNIFICANCE: Gp120 induced caspase 8 dependent NF-kappaB activation is a novel pathway of HIV replication which increases understanding of the biology of T-cell death, as well as having implications for understanding treatment and prevention of HIV infection. PMID- 19287490 TI - Genetic diversity in the SIR model of pathogen evolution. AB - We introduce a model for assessing the levels and patterns of genetic diversity in pathogen populations, whose epidemiology follows a susceptible-infected recovered model (SIR). We model the population of pathogens as a metapopulation composed of subpopulations (infected hosts), where pathogens replicate and mutate. Hosts transmit pathogens to uninfected hosts. We show that the level of pathogen variation is well predicted by analytical expressions, such that pathogen neutral molecular variation is bounded by the level of infection and increases with the duration of infection. We then introduce selection in the model and study the invasion probability of a new pathogenic strain whose fitness (R(0)(1+s)) is higher than the fitness of the resident strain (R(0)). We show that this invasion probability is given by the relative increment in R(0) of the new pathogen (s). By analyzing the patterns of genetic diversity in this framework, we identify the molecular signatures during the replacement and compare these with those observed in sequences of influenza A. PMID- 19287491 TI - The photosynthetic apparatus and its regulation in the aerobic gammaproteobacterium Congregibacter litoralis gen. nov., sp. nov. AB - BACKGROUND: There is accumulating evidence that in some marine environments aerobic bacteriochlorophyll a-producing bacteria represent a significant part of the microbial population. The interaction of photosynthesis and carbon metabolism in these interesting bacteria is still largely unknown and requires further investigation in order to estimate their contribution to the marine carbon cycle. METHODOLOGY/PRINCIPAL FINDINGS: Here, we analyzed the structure, composition and regulation of the photosynthetic apparatus in the obligately aerobic marine gammaproteobacterium KT71(T). Photoheterotrophically grown cells were characterized by a poorly developed lamellar intracytoplasmic membrane system, a type 1 light-harvesting antenna complex and a photosynthetic reaction center associated with a tetraheme cytochrome c. The only photosynthetic pigments produced were bacteriochlorophyll a and spirilloxanthin. Under semiaerobic conditions KT71(T) cells expressing a photosynthetic apparatus showed a light dependent increase of growth yield in the range of 1.3-2.5 fold. The expression level of the photosynthetic apparatus depended largely on the utilized substrate, the intermediary carbon metabolism and oxygen tension. In addition, pigment synthesis was strongly influenced by light, with blue light exerting the most significant effect, implicating that proteins containing a BLUF domain may be involved in regulation of the photosynthetic apparatus. Several phenotypic traits in KT71(T) could be identified that correlated with the assumed redox state of growing cells and thus could be used to monitor the cellular redox state under various incubation conditions. CONCLUSIONS/SIGNIFICANCE: In a hypothetical model that explains the regulation of the photosynthetic apparatus in strain KT71(T) we propose that the expression of photosynthesis genes depends on the cellular redox state and is maximal under conditions that allow a balanced membrane redox state. So far, bacteria capable of an obligately aerobic, photosynthetic metabolism constitute a unique phenotype within the class Gammaproteobacteria, so that it is justified to propose a new genus and species, Congregibacter litoralis gen. nov, sp. nov., represented by the type strain KT71(T) ( = DSM 17192(T) = NBRC 104960(T)). PMID- 19287492 TI - A conserved function of C. elegans CASY-1 calsyntenin in associative learning. AB - BACKGROUND: Whole-genome association studies in humans have enabled the unbiased discovery of new genes associated with human memory performance. However, such studies do not allow for a functional or causal testing of newly identified candidate genes. Since polymorphisms in Calsyntenin 2 (CLSTN2) showed a significant association with episodic memory performance in humans, we tested the C. elegans CLSTN2 ortholog CASY-1 for possible functions in the associative behavior of C. elegans. METHODOLOGY/PRINCIPAL FINDINGS: Using three different associative learning paradigms and functional rescue experiments, we show that CASY-1 plays an important role during associative learning in C. elegans. Furthermore, neuronal expression of human CLSTN2 in C. elegans rescues the learning defects of casy-1 mutants. Finally, genetic interaction studies and neuron-specific expression experiments suggest that CASY-1 may regulate AMPA-like GLR-1 glutamate receptor signaling. CONCLUSION/SIGNIFICANCE: Our experiments demonstrate a remarkable conservation of the molecular function of Calsyntenins between nematodes and humans and point at a role of C. elegans casy-1 in regulating a glutamate receptor signaling pathway. PMID- 19287493 TI - cGMP-dependent protein kinase I is crucial for angiogenesis and postnatal vasculogenesis. AB - BACKGROUND: Endothelium-derived nitric oxide plays an important role for the bone marrow microenvironment. Since several important effects of nitric oxide are mediated by cGMP-dependent pathways, we investigated the role of the cGMP downstream effector cGMP-dependent protein kinase I (cGKI) on postnatal neovascularization. METHODOLOGY/PRINCIPAL FINDINGS: In a disc neovascularization model, cGKI(-/-) mice showed an impaired neovascularization as compared to their wild-type (WT) littermates. Infusion of WT, but not cGKI(-/-) bone marrow progenitors rescued the impaired ingrowth of new vessels in cGKI-deficient mice. Bone marrow progenitors from cGKI(-/-) mice showed reduced proliferation and survival rates. In addition, we used cGKIalpha leucine zipper mutant (LZM) mice as model for cGKI deficiency. LZM mice harbor a mutation in the cGKIalpha leucine zipper that prevents interaction with downstream signaling molecules. Consistently, LZM mice exhibited reduced numbers of vasculogenic progenitors and impaired neovascularization following hindlimb ischemia compared to WT mice. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that the cGMP-cGKI pathway is critical for postnatal neovascularization and establish a new role for cGKI in vasculogenesis, which is mediated by bone marrow-derived progenitors. PMID- 19287494 TI - Regulation of clock-controlled genes in mammals. AB - The complexity of tissue- and day time-specific regulation of thousands of clock controlled genes (CCGs) suggests that many regulatory mechanisms contribute to the transcriptional output of the circadian clock. We aim to predict these mechanisms using a large scale promoter analysis of CCGs.Our study is based on a meta-analysis of DNA-array data from rodent tissues. We searched in the promoter regions of 2065 CCGs for highly overrepresented transcription factor binding sites. In order to compensate the relatively high GC-content of CCG promoters, a novel background model to avoid a bias towards GC-rich motifs was employed. We found that many of the transcription factors with overrepresented binding sites in CCG promoters exhibit themselves circadian rhythms. Among the predicted factors are known regulators such as CLOCKratioBMAL1, DBP, HLF, E4BP4, CREB, RORalpha and the recently described regulators HSF1, STAT3, SP1 and HNF-4alpha. As additional promising candidates of circadian transcriptional regulators PAX-4, C/EBP, EVI-1, IRF, E2F, AP-1, HIF-1 and NF-Y were identified. Moreover, GC-rich motifs (SP1, EGR, ZF5, AP-2, WT1, NRF-1) and AT-rich motifs (MEF-2, HMGIY, HNF-1, OCT-1) are significantly overrepresented in promoter regions of CCGs. Putative tissue-specific binding sites such as HNF-3 for liver, NKX2.5 for heart or Myogenin for skeletal muscle were found. The regulation of the erythropoietin (Epo) gene was analysed, which exhibits many binding sites for circadian regulators. We provide experimental evidence for its circadian regulated expression in the adult murine kidney. Basing on a comprehensive literature search we integrate our predictions into a regulatory network of core clock and clock-controlled genes. Our large scale analysis of the CCG promoters reveals the complexity and extensiveness of the circadian regulation in mammals. Results of this study point to connections of the circadian clock to other functional systems including metabolism, endocrine regulation and pharmacokinetics. PMID- 19287495 TI - Association and haplotype analyses of positional candidate genes in five genomic regions linked to scrotal hernia in commercial pig lines. AB - Scrotal hernia in pigs is a complex trait likely affected by genetic and environmental factors. A large-scale association analysis of positional and functional candidate genes was conducted in four previously identified genomic regions linked to hernia susceptibility on Sus scrofa chromosomes 2 and 12, as well as the fifth region around 67 cM on chromosome 2, respectively. In total, 151 out of 416 SNPs discovered were genotyped successfully. Using a family-based analysis we found that four regions surrounding ELF5, KIF18A, COL23A1 on chromosome 2, and NPTX1 on chromosome 12, respectively, may contain the genetic variants important for the development of the scrotal hernia in pigs. These findings were replicated in another case-control dataset. The SNPs around the ELF5 region were in high linkage disequilibrium with each other, and a haplotype containing SNPs from ELF5 and CAT was highly significantly associated with hernia development. Extensive re-sequencing work focused on the KIF18A gene did not detect any further SNPs with extensive association signals. These genes may be involved in the estrogen receptor signaling pathway (KIF18A and NPTX1), the epithelial-mesenchymal transition (ELF5) and the collagen metabolism pathway (COL23A1), which are associated with the important molecular characteristics of hernia pathophysiology. Further investigation on the molecular mechanisms of these genes may provide more molecular clues on hernia development in pigs. PMID- 19287496 TI - FOXM1 upregulation is an early event in human squamous cell carcinoma and it is enhanced by nicotine during malignant transformation. AB - BACKGROUND: Cancer associated with smoking and drinking remains a serious health problem worldwide. The survival of patients is very poor due to the lack of effective early biomarkers. FOXM1 overexpression is linked to the majority of human cancers but its mechanism remains unclear in head and neck squamous cell carcinoma (HNSCC). METHODOLOGY/PRINCIPAL FINDINGS: FOXM1 mRNA and protein expressions were investigated in four independent cohorts (total 75 patients) consisting of normal, premalignant and HNSCC tissues and cells using quantitative PCR (qPCR), expression microarray, immunohistochemistry and immunocytochemistry. Effect of putative oral carcinogens on FOXM1 transcriptional activity was dose dependently assayed and confirmed using a FOXM1-specific luciferase reporter system, qPCR, immunoblotting and short-hairpin RNA interference. Genome-wide single nucleotide polymorphism (SNP) array was used to 'trace' the genomic instability signature pattern in 8 clonal lines of FOXM1-induced malignant human oral keratinocytes. Furthermore, acute FOXM1 upregulation in primary oral keratinocytes directly induced genomic instability. We have shown for the first time that overexpression of FOXM1 precedes HNSCC malignancy. Screening putative carcinogens in human oral keratinocytes surprisingly showed that nicotine, which is not perceived to be a human carcinogen, directly induced FOXM1 mRNA, protein stabilisation and transcriptional activity at concentrations relevant to tobacco chewers. Importantly, nicotine also augmented FOXM1-induced transformation of human oral keratinocytes. A centrosomal protein CEP55 and a DNA helicase/putative stem cell marker HELLS, both located within a consensus loci (10q23), were found to be novel targets of FOXM1 and their expression correlated tightly with HNSCC progression. CONCLUSIONS/SIGNIFICANCE: This study cautions the potential co carcinogenic effect of nicotine in tobacco replacement therapies. We hypothesise that aberrant upregulation of FOXM1 may be inducing genomic instability through a program of malignant transformation involving the activation of CEP55 and HELLS which may facilitate aberrant mitosis and epigenetic modifications. Our finding that FOXM1 is upregulated early during oral cancer progression renders FOXM1 an attractive diagnostic biomarker for early cancer detection and its candidate mechanistic targets, CEP55 and HELLS, as indicators of malignant conversion and progression. PMID- 19287497 TI - Blood glucose levels regulate pancreatic beta-cell proliferation during experimentally-induced and spontaneous autoimmune diabetes in mice. AB - BACKGROUND: Type 1 diabetes mellitus is caused by immune-mediated destruction of pancreatic beta-cells leading to insulin deficiency, impaired intermediary metabolism, and elevated blood glucose concentrations. While at autoimmune diabetes onset a limited number of beta-cells persist, the cells' regenerative potential and its regulation have remained largely unexplored. Using two mouse autoimmune diabetes models, this study examined the proliferation of pancreatic islet ss-cells and other endocrine and non-endocrine subsets, and the factors regulating that proliferation. METHODOLOGY AND PRINCIPAL FINDINGS: We adapted multi-parameter flow cytometry techniques (including DNA-content measurements and 5'-bromo-2'-deoxyuridine [BrdU] incorporation) to study pancreatic islet single cell suspensions. These studies demonstrate that beta-cell proliferation rapidly increases at diabetes onset, and that this proliferation is closely correlated with the diabetic animals' elevated blood glucose levels. For instance, we show that when normoglycemia is restored by exogenous insulin or islet transplantation, the beta-cell proliferation rate returns towards low levels found in control animals, yet surges when hyperglycemia recurs. In contrast, other-than-ss endocrine islet cells did not exhibit the same glucose-dependent proliferative responses. Rather, disease-associated alterations of BrdU incorporation rates of delta-cells (minor decrease), and non-endocrine islet cells (slight increase) were not affected by blood glucose levels, or were inversely related to glycemia control after diabetes onset (alpha-cells). CONCLUSION: We conclude that murine beta-cells' ability to proliferate in response to metabolic need (i.e. rising blood glucose concentrations) is remarkably well preserved during severe, chronic beta-cell autoimmunity. These data suggest that timely control of the destructive immune response after disease manifestation could allow spontaneous regeneration of sufficient beta-cell mass to restore normal glucose homeostasis. PMID- 19287498 TI - PTCH1 +/- dermal fibroblasts isolated from healthy skin of Gorlin syndrome patients exhibit features of carcinoma associated fibroblasts. AB - Gorlin's or nevoid basal cell carcinoma syndrome (NBCCS) causes predisposition to basal cell carcinoma (BCC), the commonest cancer in adult human. Mutations in the tumor suppressor gene PTCH1 are responsible for this autosomal dominant syndrome. In NBCCS patients, as in the general population, ultraviolet exposure is a major risk factor for BCC development. However these patients also develop BCCs in sun protected areas of the skin, suggesting the existence of other mechanisms for BCC predisposition in NBCCS patients. As increasing evidence supports the idea that the stroma influences carcinoma development, we hypothesized that NBCCS fibroblasts could facilitate BCC occurence of the patients. WT (n = 3) and NBCCS fibroblasts bearing either nonsense (n = 3) or missense (n = 3) PTCH1 mutations were cultured in dermal equivalents made of a collagen matrix and their transcriptomes were compared by whole genome microarray analyses. Strikingly, NBCCS fibroblasts over-expressed mRNAs encoding pro-tumoral factors such as Matrix Metalloproteinases 1 and 3 and tenascin C. They also over-expressed mRNA of pro-proliferative diffusible factors such as fibroblast growth factor 7 and the stromal cell-derived factor 1 alpha, known for its expression in carcinoma associated fibroblasts. These data indicate that the PTCH1(+/-) genotype of healthy NBCCS fibroblasts results in phenotypic traits highly reminiscent of those of BCC associated fibroblasts, a clue to the yet mysterious proneness to non photo-exposed BCCs in NBCCS patients. PMID- 19287499 TI - Frequency and interrelations of risk factors for chronic low back pain in a primary care setting. AB - INTRODUCTION: Many risk factors have been identified for chronic low back pain (cLBP), but only one study evaluated their interrelations. We aimed to investigate the frequency of cLBP risk factors and their interrelations in patients consulting their general practitioners (GPs) for cLBP. METHODS: A cross sectional, descriptive, national survey was performed. 3000 GPs randomly selected were asked to include at least one patient consulting for cLBP. Demographic, clinical characteristics and the presence of cLBP risk factors were recorded. The frequency of each cLBP risk factor was calculated and multiple correspondence analysis (MCA) was performed to study their interrelations. RESULTS: A total of 2068 GPs (68.9%) included at least 1 patient, for 4522 questionnaires analyzed. In the whole sample of patients, the 2 risk factors most commonly observed were history of recurrent LBP (72.1%) and initial limitation of activities of daily living (66.4%). For working patients, common professional risk factors were beliefs, that LBP was due to maintaining a specific posture at work (79.0%) and frequent heavy lifting at work (65.5%). On MCA, we identified 3 risk-factor dimensions (axes) for working and nonworking patients. The main dimension for working patients involved professional risk factors and among these factors, patients' job satisfaction and job recognition largely contribute to this dimension. DISCUSSION: Our results shed in light for the first time the interrelation and the respective contribution of several previously identified cLBP risk factors. They suggest that risk factors representing a "work-related" dimension are the most important cLBP risk factors in the working population. PMID- 19287500 TI - Redistribution of actin during assembly and reassembly of the contractile ring in grasshopper spermatocytes. AB - Cytokinesis in animal cells requires the assembly of an actomyosin contractile ring to cleave the cell. The ring is highly dynamic; it assembles and disassembles during each cell cleavage, resulting in the recurrent redistribution of actin. To investigate this process in grasshopper spermatocytes, we mechanically manipulated the spindle to induce actin redistribution into ectopic contractile rings, around reassembled lateral spindles. To enhance visualization of actin, we folded the spindle at its equator to convert the remnants of the partially assembled ring into a concentrated source of actin. Filaments from the disintegrating ring aligned along reorganizing spindle microtubules, suggesting that their incorporation into the new ring was mediated by microtubules. We tracked incorporation by speckling actin filaments with Qdots and/or labeling them with Alexa 488-phalloidin. The pattern of movement implied that actin was transported along spindle microtubules, before entering the ring. By double labeling dividing cells, we imaged actin filaments moving along microtubules near the contractile ring. Together, our findings indicate that in one mechanism of actin redistribution, actin filaments are transported along spindle microtubule tracks in a plus-end-directed fashion. After reaching the spindle midzone, the filaments could be transported laterally to the ring. Notably, actin filaments undergo a dramatic trajectory change as they enter the ring, implying the existence of a pulling force. Two other mechanisms of actin redistribution, cortical flow and de novo assembly, are also present in grasshopper, suggesting that actin converges at the nascent contractile ring from diffuse sources within the cytoplasm and cortex, mediated by spindle microtubules. PMID- 19287501 TI - Upregulation of alpha7 Nicotinic Receptors by Acetylcholinesterase C-Terminal Peptides. AB - BACKGROUND: The alpha-7 nicotinic acetylcholine receptor (alpha7-nAChR) is well known as a potent calcium ionophore that, in the brain, has been implicated in excitotoxicity and hence in the underlying mechanisms of neurodegenerative disorders such as Alzheimer's disease. Previous research implied that the activity of this receptor may be modified by exposure to a peptide fragment derived from the C-terminal region of the enzyme acetylcholinesterase. This investigation was undertaken to determine if the functional changes observed could be attributed to peptide binding interaction with the alpha7-nAChR, or peptide modulation of receptor expression. METHODOLOGY/PRINCIPAL FINDINGS: This study provides evidence that two peptides derived from the C-terminus of acetylcholinesterase, not only selectively displace specific bungarotoxin binding at the alpha7-nAChR, but also alter receptor binding properties for its familiar ligands, including the alternative endogenous agonist choline. Of more long-term significance, these peptides also induce upregulation of alpha7-nAChR mRNA and protein expression, as well as enhancing receptor trafficking to the plasma membrane. CONCLUSIONS/SIGNIFICANCE: The results reported here demonstrate a hitherto unknown relationship between the alpha7-nAChR and the non-enzymatic functions of acetylcholinesterase, mediated independently by its C-terminal domain. Such an interaction may prove valuable as a pharmacological tool, prompting new approaches for understanding, and combating, the process of neurodegeneration. PMID- 19287502 TI - RBF-TSS: identification of transcription start site in human using radial basis functions network and oligonucleotide positional frequencies. AB - Accurate identification of promoter regions and transcription start sites (TSS) in genomic DNA allows for a more complete understanding of the structure of genes and gene regulation within a given genome. Many recently published methods have achieved high identification accuracy of TSS. However, models providing more accurate modeling of promoters and TSS are needed. A novel identification method for identifying transcription start sites that improves the accuracy of TSS recognition for recently published methods is proposed. This method incorporates a metric feature based on oligonucleotide positional frequencies, taking into account the nature of promoters. A radial basis function neural network for identifying transcription start sites (RBF-TSS) is proposed and employed as a classification algorithm. Using non-overlapping chunks (windows) of size 50 and 500 on the human genome, the proposed method achieves an area under the Receiver Operator Characteristic curve (auROC) of 94.75% and 95.08% respectively, providing increased performance over existing TSS prediction methods. PMID- 19287503 TI - Phosphorylation modification of wheat lectin VER2 is associated with vernalization-induced O-GlcNAc signaling and intracellular motility. AB - BACKGROUND: O-linked beta-N-acetylglucosamine (O-GlcNAc) modification of proteins mediates stress response and cellular motility in animal cells. The plant lectin concanavalin A can increase nuclear O-GlcNAc levels and decrease cytoplasmic O GlcNAc levels in T lymphocytes. However, the functions of O-GlcNAc signaling in plants, as well as the relation between plant lectins and O-GlcNAc in response to environmental stimuli are largely undefined. METHODOLOGY/PRINCIPAL FINDINGS: We describe a jacalin-like lectin VER2 in wheat that shows N-acetylglucosamine and galactose specificity. Immunocytochemical localization showed VER2 expression induced predominantly at potential nuclear structures in shoot tips and young leaves and weakly in cytoplasm in response to vernalization. In contrast, under devernalization (continuous stimulation with a higher temperature after vernalization), VER2 signals appeared predominantly in cytoplasm. 2-D electrophoresis, together with western blot analysis, showed phosphorylation modification of VER2 under vernalization. Immunoblot assay with O-GlcNAc-specific antibody revealed that vernalization increased O-GlcNAc modification of proteins at the global level. An O-GlcNAc-modified protein co-immunoprecipitated with VER2 in vernalized wheat plants but not in devernalized materials. The dynamic of VER2 was observed in transgenic Arabidopsis overexpressing the VER2-GFP fusion protein. Overexpressed VER2 accelerated nuclear migration. Immunogold labeling and indirect immunofluoresence colocalization assay indicated that VER2-GFP was targeted to the secretory pathway. CONCLUSIONS/SIGNIFICANCE: O-GlcNAc signaling is involved in the vernalization response in wheat, and phosphorylation is necessary for the lectin VER2 involving O-GlcNAc signaling during vernalization. Our findings open the way to studies of O-GlcNAc protein modification in response to environmental signals in plants. PMID- 19287504 TI - Evaluation and characterization of bacterial metabolic dynamics with a novel profiling technique, real-time metabolotyping. AB - BACKGROUND: Environmental processes in ecosystems are dynamically altered by several metabolic responses in microorganisms, including intracellular sensing and pumping, battle for survival, and supply of or competition for nutrients. Notably, intestinal bacteria maintain homeostatic balance in mammals via multiple dynamic biochemical reactions to produce several metabolites from undigested food, and those metabolites exert various effects on mammalian cells in a time dependent manner. We have established a method for the analysis of bacterial metabolic dynamics in real time and used it in combination with statistical NMR procedures. METHODOLOGY/PRINCIPAL FINDINGS: We developed a novel method called real-time metabolotyping (RT-MT), which performs sequential (1)H-NMR profiling and two-dimensional (2D) (1)H, (13)C-HSQC (heteronuclear single quantum coherence) profiling during bacterial growth in an NMR tube. The profiles were evaluated with such statistical methods as Z-score analysis, principal components analysis, and time series of statistical TOtal Correlation SpectroScopY (TOCSY). In addition, using 2D (1)H, (13)C-HSQC with the stable isotope labeling technique, we observed the metabolic kinetics of specific biochemical reactions based on time-dependent 2D kinetic profiles. Using these methods, we clarified the pathway for linolenic acid hydrogenation by a gastrointestinal bacterium, Butyrivibrio fibrisolvens. We identified trans11, cis13 conjugated linoleic acid as the intermediate of linolenic acid hydrogenation by B. fibrisolvens, based on the results of (13)C-labeling RT-MT experiments. In addition, we showed that the biohydrogenation of polyunsaturated fatty acids serves as a defense mechanism against their toxic effects. CONCLUSIONS: RT-MT is useful for the characterization of beneficial bacterium that shows potential for use as probiotic by producing bioactive compounds. PMID- 19287505 TI - Understanding the transition to acute illness: the promise of perioperative genomics. AB - Over the last decades, tremendous advances have been made in understanding the genomic basis of a large number of chronic human diseases. Such advances were made possible by studying large numbers of patients over relatively long periods of time. It is increasingly recognized that acute robust stress stimulates pathways activated in chronic disease, facilitating mechanistic studies in much shorter time frames. A new field of molecular medicine, called perioperative genomics, uses robust surgical stress as a perturbation designed to explore such mechanisms. This new field is described briefly below. PMID- 19287506 TI - HIV Sexual Risk Behaviors among Ketamine and Non-Ketamine Using Criminal Offenders Prior to Prison Entry. AB - This study is the first to examine ketamine use and its association with HIV sexual risk behaviors among a criminal offending population in the United States. Data were collected from 716 inmates as part of the Transitional Case Management (TCM) protocol within the Criminal Justice Drug Abuse Treatment Studies (CJ-DATS) cooperative agreement. Bivariate analyses were used to identify differences between ketamine users (n=44) and non-ketamine users (n=672). Three Poisson regression models were used to identify the significant correlates of high risk sexual behaviors in the 30 days prior to incarceration-- (1) number of times had unprotected sex while high, (2) number of times had unprotected vaginal sex, and (3) number of times had unprotected anal sex. Results indicate that ketamine was a significant correlate in all of the Poisson regression models. Findings add to the literature and indicate that ketamine use may be a marker for engaging in HIV risk behaviors among criminal offenders. PMID- 19287507 TI - Exact and approximate cone-beam reconstruction algorithms for C-arm based cone beam CT using a two-concentric-arc source trajectory. AB - In this paper, we present shift-invariant filtered backprojection (FBP) cone-beam image reconstruction algorithms for a cone-beam CT system based on a clinical C arm gantry. The source trajectory consists of two concentric arcs which is complete in the sense that the Tuy data sufficiency condition is satisfied. This scanning geometry is referred to here as a CC geometry (each arc is shaped like the letter "C"). The challenge for image reconstruction for the CC geometry is that the image volume is not well populated by the familiar doubly measured (DM) lines. Thus, the well-known DM-line based image reconstruction schemes are not appropriate for the CC geometry. Our starting point is a general reconstruction formula developed by Pack and Noo which is not dependent on the existence of DM lines. For a specific scanning geometry, the filtering lines must be carefully selected to satisfy the Pack-Noo condition for mathematically exact reconstruction. The new points in this paper are summarized here. (1) A mathematically exact cone-beam reconstruction algorithm was formulated for the CC geometry by utilizing the Pack-Noo image reconstruction scheme. One drawback of the developed exact algorithm is that it does not solve the long-object problem. (2) We developed an approximate image reconstruction algorithm by deforming the filtering lines so that the long object problem is solved while the reconstruction accuracy is maintained. (3) In addition to numerical phantom experiments to validate the developed image reconstruction algorithms, we also validate our algorithms using physical phantom experiments on a clinical C-arm system. PMID- 19287508 TI - Dual localization of wild-type myocilin in the endoplasmic reticulum and extracellular compartment likely occurs due to its incomplete secretion. AB - PURPOSE: Wild-type myocilin is known to be secreted extracellularly, but a significant amount of the protein is also present in the endoplasmic reticulum (ER). The present study was undertaken to address whether intracellular myocilin is a true ER resident protein. METHODS: Human wild-type myocilin was adenovirally expressed in human trabecular meshwork cells, and general characteristics of both intracellular and extracellular myocilins including molecular weight, pI, glycosylation state, and cleavage site of the signal peptide were examined by biochemical analyses. Topology and decay kinetics of myocilin were also examined by protease protection assay and pulse chase analysis, respectively. The expression pattern and cytopathic effect of myocilin were analyzed in individual cells by immunocytochemistry. RESULTS: Intracellular myocilin were very similar to secreted myocilin in characteristics such as molecular weight, pI, glycosylation state, and cleavage site of the signal peptide. The intracellular protein was found to be present in the lumen of the ER where it appeared to be retained without further export to the Golgi apparatus. The kinetics of myocilin turnover clearly showed that it was intrinsically a very stable but incompletely secreted protein. The expression of myocilin was confined to a subset of cells and accompanied by the upregulation of a 78 kDa glucose-regulated protein, suggesting that it was not properly folded or processed in the ER. CONCLUSIONS: Based on these findings and the fact that myocilin has no known ER retention signals, the ER localization of wild-type myocilin is likely a consequence of its incomplete secretion due to its misfolding. PMID- 19287509 TI - Genome-wide association study of determinants of anti-cyclic citrullinated peptide antibody titer in adults with rheumatoid arthritis. AB - We carried out a genome-wide association study of genetic predictors of anti cyclic citrullinated peptide antibody (anti-CCP) level in 531 self-reported non Hispanic Caucasian Rheumatoid Arthritis (RA) patients enrolled in the Brigham Rheumatoid Arthritis Sequential Study (BRASS). For replication, we then analyzed 289 single nucleotide polymorphisms (SNPs) with P < 0.001 in BRASS in an independent population of 849 RA patients from the North American Rheumatoid Arthritis Consortium (NARAC). BRASS and NARAC samples were genotyped using the Affymetrix 100K and Illumina 550K platforms respectively. Association between SNPs and anti-CCP titer was tested using general linear models. The five most significant SNPs from BRASS all were within the major histocompatibility complex (MHC) region (P < or = 3.5 x 10(-6)). After controlling for the human leukocyte antigen shared epitope (HLA-SE), the top SNPs still yielded P values < 0.0002. In NARAC, a single SNP from the MHC region near BTNL2 and HLA-DRA, rs1980493 (r(2) = 0.85 with the top five SNPs from BRASS), was associated significantly with CCP titer (P = 6.1 x 10(-5)) even after adjustment for the HLA-SE (P = 0.0002). The top SNPs found in BRASS and NARAC had r(2) = 0.46 and 0.64, respectively, to HLA DRB1 DR3 alleles. These results confirm that the most significant genome region affecting anti-CCP titers in RA is the MHC region. We identified a SNP in moderate linkage disequilibrium (LD) with HLA-DR3, which may influence anti-CCP titer independently of the HLA-SE. PMID- 19287510 TI - A novel recombinant vaccinia virus expressing the human norepinephrine transporter retains oncolytic potential and facilitates deep-tissue imaging. AB - Noninvasive and repetitive monitoring of a virus in target tissues and/or specific organs of the body is highly desirable for the development of safe and efficient cancer virotherapeutics. We have previously shown that the oncolytic vaccinia virus GLV-1h68 can target and eradicate human tumors in mice and that its therapeutic effects can be monitored by using optical imaging. Here, we report on the development of a derivative of GLV-1h68, a novel recombinant vaccinia virus (VACV) GLV-1h99, which was constructed to carry the human norepinephrine transporter gene (hNET) under the VACV synthetic early promoter placed at the F14.5L locus for deep-tissue imaging. The hNET protein was expressed at high levels on the membranes of cells infected with this virus. Expression of the hNET protein did not negatively affect virus replication, cytolytic activity in cell culture, or in vivo virotherpeutic efficacy. GLV-1h99 mediated expression of the hNET protein in infected cells resulted in specific uptake of the radiotracer [131I]-meta-iodobenzylguanidine (MIBG). In mice, GLV 1h99-infected tumors were readily imaged by [124I]-MIBG positron emission tomography. To our knowledge, GLV-1h99 is the first oncolytic virus expressing the hNET protein that can efficiently eliminate tumors and simultaneously allow deep-tissue imaging of infected tumors. PMID- 19287511 TI - Analysis association between mitochondrial genome instability and xenobiotic metabolizing genes in human breast cancer. AB - The aim of this study was to determine the existence of association between the genetic polymorphisms of metabolizing genes GSTM-1, GSTT-1, and NAT-2, and the presence of mitochondrial genome instability (mtGI) in breast cancer cases. Ninety-four pairs of tumoral/nontumoral breast cancer samples were analyzed. Our samples showed 40.42% of mtGI by analysis of two D-loop region markers, a (CA)n mtMS starting at the 514-bp position, and four informative MnlI sites between the 16,108-16,420-bp. GSTM-1 null genotype has shown a significant association with mtGI presence (chi(2) = 7.62; P = 0.006) in breast cancer cases; moreover, these genotypes also are related to an increased risk for mtDNA damage (odds ratio [OR] = 3.71 [1.41-9.88]; 95% Cornfield confidence interval [CI]). These results suggest that the absence of GSTM-1 enzymatic activity favors chemical actions in damaging the mtDNA. Analysis of GSTT-1 and NAT-2 polymorphisms showed no association with mtGI (chi(2) = 0.03; P = 0.87 and chi(2) = 2.76; P = 0.09, respectively). The analysis of invasive breast cancer cases showed mtGI in 74.36% of ILC cases (29 of 39 samples), and in only 18.75% (9 out of 48) IDC cases; this result suggests a possible relation between mtDNA mutations and variations in molecular pathways of tumor development. PMID- 19287512 TI - Oxygen-conserving implications of the trigemino-cardiac reflex in the brain: the molecular basis of neuroprotection? PMID- 19287523 TI - Drought meets acid: three new genera in a dothidealean clade of extremotolerant fungi. AB - Fungal strains isolated from rocks and lichens collected in the Antarctic ice free area of the Victoria Land, one of the coldest and driest habitats on earth, were found in two phylogenetically isolated positions within the subclass Dothideomycetidae. They are here reported as new genera and species, Recurvomyces mirabilisgen. nov., sp. nov. and Elasticomyces elasticusgen. nov., sp. nov. The nearest neighbours within the clades were other rock-inhabiting fungi from dry environments, either cold or hot. Plant-associated Mycosphaerella-like species, known as invaders of leathery leaves in semi-arid climates, are also phylogenetically related with the new taxa. The clusters are also related to the halophilic species Hortaea werneckii, as well as to acidophilic fungi. One of the latter, able to grow at pH 0, is Scytalidium acidophilum, which is ascribed here to the newly validated genus Acidomyces. The ecological implications of this finding are discussed. PMID- 19287524 TI - Redefinition of Aureobasidium pullulans and its varieties. AB - Using media with low water activity, a large numbers of aureobasidium-like black yeasts were isolated from glacial and subglacial ice of three polythermal glaciers from the coastal Arctic environment of Kongsfjorden (Svalbard, Spitsbergen), as well as from adjacent sea water, sea ice and glacial meltwaters. To characterise the genetic variability of Aureobasidium pullulans strains originating from the Arctic and strains originating pan-globally, a multilocus molecular analysis was performed, through rDNA (internal transcribed spacers, partial 28 S rDNA), and partial introns and exons of genes encoding beta-tubulin (TUB), translation elongation factor (EF1alpha) and elongase (ELO). Two globally ubiquitous varieties were distinguished: var. pullulans, occurring particularly in slightly osmotic substrates and in the phyllosphere; and var. melanogenum, mainly isolated from watery habitats. Both varieties were commonly isolated from the sampled Arctic habitats. However, some aureobasidium-like strains from subglacial ice from three different glaciers in Kongsfjorden (Svalbard, Spitsbergen), appeared to represent a new variety of A. pullulans. A strain from dolomitic marble in Namibia was found to belong to yet another variety. No molecular support has as yet been found for the previously described var. aubasidani. A partial elongase-encoding gene was successfully used as a phylogenetic marker at the (infra-)specific level. PMID- 19287525 TI - The influence of ortho- and para-diphenoloxidase substrates on pigment formation in black yeast-like fungi. AB - Dothideaceous black yeast-like fungi (BYF) are known to synthesise DHN-melanin that is inhibited by the systemic fungicide tricyclazole. The final step of the DHN melanin pathway is the conjoining of 1,8-DHN molecules to form the melanin polymer. There are several candidate enzymes for this step, including phenoloxidases such as tyrosinase and laccases, peroxidases, and perhaps also catalases. We analysed the type polyphenoloxidases that are involved in biosynthesis of BYF melanins. For that purpose we used substrates of o diphenoloxidases (EC 1.10.3.1.): 4-hydroxyphenyl-pyruvic acid, L-beta phenyllactic acid, tyrosine, pyrocatechol, 3,4-dihydroxyphenylalanine and homogentisic acid, as well as substrates of p-diphenoloxidases (EC 1.10.3.2.): syringaldazine, resorcinol, p-phenylenediamine, phloroglucinol, guaiacol and pyrogallic acid. Fourteen strains of black yeasts originating from different natural biotopes were investigated. The tested strains could be divided into four groups based on their ability to produce dark pigments when cultivated on aromatic substrates of o- and on p-diphenoloxidases. It was established that syringaldazine, pyrogallic acid and 4-hydrophenyl-pyruvic acid, beta-phenyllactic acid optimally promote melanin biosynthesis. Average intensity of pigmentation of all strains studied was minimal when guaiacol was used as a substrate. The present investigation indicates that the melanisation process may involve more enzymes and more substrates than those commonly recognised. Black yeasts are likely to contain a multipotent polyphenoloxidase. PMID- 19287526 TI - Expression of fatty-acid-modifying enzymes in the halotolerant black yeast Aureobasidium pullulans (de Bary) G. Arnaud under salt stress. AB - Multiple tolerance to stressful environmental conditions of the black, yeast-like fungus Aureobasidium pullulans is achieved through different adaptations, among which there is the restructuring of the lipid composition of their membranes. Here, we describe three novel genes encoding fatty-acid-modifying enzymes in A. pullulans, along with the levels of their mRNAs under different salinity conditions. High levels of Delta(9)-desaturase and Delta(12)-desaturase mRNAs were seen at high salinities, which were consistent with an increased desaturation of the fatty acids in the cell membranes. Elevated levels of elongase mRNA were also detected. Surprisingly, increases in the levels of these mRNAs were also seen following hypo-osmotic shock, while hyperosmotic shock had exactly the opposite effect, demonstrating that data that are obtained from up shift and down-shift salinity studies should be interpreted with caution. PMID- 19287527 TI - HMG-CoA reductase is regulated by environmental salinity and its activity is essential for halotolerance in halophilic fungi. AB - The activity and level of HMG-CoA reductase (HMGR) were addressed in halophilic fungi isolated from solar saltpans. Representative fungi belonging to the orders Dothideales, Eurotiales and Wallemiales have a specific pattern of HMGR regulation, which differs from salt-sensitive and moderately salt-tolerant yeasts. In all of the halophilic fungi studied, HMGR amounts and activities were the lowest at optimal growth salinity and increased under hyposaline and hypersaline conditions. This profile paralleled isoprenylation of cellular proteins in H. werneckii. Inhibition of HMGR in vivo by lovastatin impaired the halotolerant character. HMGR may thus serve as an important molecular marker of halotolerance. PMID- 19287529 TI - Tinea nigra by Hortaea werneckii, a report of 22 cases from Mexico. AB - Tinea nigra is a superficial mycosis caused by Hortaea werneckii. It is an infrequent asymptomatic infection that affects human palms and soles, and is mostly observed in tropical countries. We evaluate retrospectively twenty-two confirmed cases of tinea nigra from a total of eleven yr (1997-2007) and discuss the epidemiology, clinical features and treatment of this disease. In twelve cases, adults were involved, in 10, children. In nineteen cases the disorder was located on palms of hands and in three on soles of feet. In all cases, the obtained isolates were morphologically identified as Hortaea werneckii and the identification of ten isolates was retrospectively confirmed with the help of sequences of the internal transcribed spacer regions of the ribosomal DNA. The patients received topical treatment with Whitfield ointment, ketoconazole, bifonazole, or terbinafine. Treatment with keratolytic agents and topical antifungals was effective. PMID- 19287528 TI - Adaptation of extremely halotolerant black yeast Hortaea werneckii to increased osmolarity: a molecular perspective at a glance. AB - Halophilic adaptations have been studied almost exclusively on prokaryotic microorganisms. Discovery of the black yeast Hortaea werneckii as the dominant fungal species in hypersaline waters enabled the introduction of a new model organism to study the mechanisms of salt tolerance in eukaryotes. Its strategies of cellular osmotic adaptations on the physiological and molecular level revealed novel, intricate mechanisms to combat fluctuating salinity. H. werneckii is an extremely halotolerant eukaryotic microorganism and thus a promising source of transgenes for osmotolerance improvement of industrially important yeasts, as well as in crops. PMID- 19287530 TI - Black fungi in lichens from seasonally arid habitats. AB - We present a phylogenetic study of black fungi in lichens, primarily focusing on saxicolous samples from seasonally arid habitats in Armenia, but also with examples from other sites. Culturable strains of lichen-associated black fungi were obtained by isolation from surface-washed lichen material. Determination is based on ITS rDNA sequence data and comparison with published sequences from other sources. The genera Capnobotryella, Cladophialophora, Coniosporium, Mycosphaerella, and Rhinocladiella were found in different lichen species, which showed no pathogenic symptoms. A clade of predominantly lichen-associated strains is present only in Rhinocladiella, whereas samples of the remaining genera were grouped more clearly in clades with species from other sources. The ecology of most-closely related strains indicates that Capnobotryella and Coniosporium, and perhaps also Rhinocladiella strains opportunistically colonise lichens. In contrast, high sequence divergence in strains assigned to Mycosphaerella could indicate the presence of several lichen-specific species with unknown range of hosts or habitats, which are distantly related to plant-inhabitants. Similar applies to Cladophialophora strains, where the closest relatives of the strains from lichens are serious human pathogens. PMID- 19287531 TI - Cellular responses of microcolonial rock fungi to long-term desiccation and subsequent rehydration. AB - Melanised rock-inhabiting fungi are astonishingly resistant to environmental stresses. Also known as micro-colonial fungi (MCF), they are ubiquitous and even colonise bare rocks in deserts. To survive in nutrient poor and extremely stressful conditions, MCF have reduced morphogenetic complexity to a minimum, and rely on a broad spectrum of stress protection mechanisms. Although visual signs of carotenoid presence are masked by heavily melanised black cell-walls, we were able to isolate and characterise a variety of carotenoids (ss-carotene, zeta carotene, phytoene, torularhodin and torulene) in the rock-inhabiting, relatively fast-growing strain A95. The desiccation/rehydration stress response was used to measure the ability of A95 to adapt to slow or fast changes in external conditions. Revival of MCF after prolonged desiccation and rehydration was documented by biochemical (analyses of lipids and protective pigments), cultivation, and microscopic methods. Survival of MCF is enhanced when desiccation is rapid and mycostasis is instant rather than following prolonged periods of low metabolic activity. PMID- 19287532 TI - Resistance of Antarctic black fungi and cryptoendolithic communities to simulated space and Martian conditions. AB - Dried colonies of the Antarctic rock-inhabiting meristematic fungi Cryomyces antarcticus CCFEE 515, CCFEE 534 and C. minteri CCFEE 5187, as well as fragments of rocks colonized by the Antarctic cryptoendolithic community, were exposed to a set of ground-based experiment verification tests (EVTs) at the German Aerospace Center (DLR, Koln, Germany). These were carried out to test the tolerance of these organisms in view of their possible exposure to space conditions outside of the International Space Station (ISS). Tests included single or combined simulated space and Martian conditions. Responses were analysed both by cultural and microscopic methods. Thereby, colony formation capacities were measured and the cellular viability was assessed using live/dead dyes FUN 1 and SYTOX Green. The results clearly suggest a general good resistance of all the samples investigated. C. minteri CCFEE 5187, C. antarcticus CCFEE 515 and colonized rocks were selected as suitable candidates to withstand space flight and long-term permanence in space on the ISS in the framework of the LIchens and Fungi Experiments (LIFE programme, European Space Agency). PMID- 19287533 TI - A rock-inhabiting ancestor for mutualistic and pathogen-rich fungal lineages. AB - Rock surfaces are unique terrestrial habitats in which rapid changes in the intensity of radiation, temperature, water supply and nutrient availability challenge the survival of microbes. A specialised, but diverse group of free living, melanised fungi are amongst the persistent settlers of bare rocks. Multigene phylogenetic analyses were used to study relationships of ascomycetes from a variety of substrates, with a dataset including a broad sampling of rock dwellers from different geographical locations. Rock-inhabiting fungi appear particularly diverse in the early diverging lineages of the orders Chaetothyriales and Verrucariales. Although these orders share a most recent common ancestor, their lifestyles are strikingly different. Verrucariales are mostly lichen-forming fungi, while Chaetothyriales, by contrast, are best known as opportunistic pathogens of vertebrates (e.g. Cladophialophora bantiana and Exophiala dermatitidis, both agents of fatal brain infections) and saprophytes. The rock-dwelling habit is shown here to be key to the evolution of these two ecologically disparate orders. The most recent common ancestor of Verrucariales and Chaetothyriales is reconstructed as a non-lichenised rock-inhabitant. Ancestral state reconstructions suggest Verrucariales as one of the independent ascomycetes group where lichenisation has evolved on a hostile rock surface that might have favored this shift to a symbiotic lifestyle. Rock-inhabiting fungi are also ancestral to opportunistic pathogens, as they are found in the early diverging lineages of Chaetothyriales. In Chaetothyriales and Verrucariales, specific morphological and physiological traits (here referred to as extremotolerance) evolved in response to stresses in extreme conditions prevailing on rock surfaces. These factors facilitated colonisation of various substrates including the brains of vertebrates by opportunistic fungal pathogens, as well as helped establishment of a stable lichen symbiosis. PMID- 19287534 TI - Evolution of CDC42, a putative virulence factor triggering meristematic growth in black yeasts. AB - The cell division cycle gene (CDC42) controlling cellular polarization was studied in members of Chaetothyriales. Based on ribosomal genes, ancestral members of the order exhibit meristematic growth in view of their colonization of inert surfaces such as rock, whereas in derived members of the order the gene is a putative virulence factor involved in expression of the muriform cell, the invasive phase in human chromoblastomycosis. Specific primers were developed to amplify a portion of the gene of 32 members of the order with known position according to ribosomal phylogeny. Phylogeny of CDC42 proved to be very different. In all members of Chaetohyriales the protein sequence is highly conserved. In most species, distributed all over the phylogenetic tree, introns and 3(rd) codon positions are also invariant. However, a number of species had paralogues with considerable deviation in non-coding exon positions, and synchronous variation in introns, although non-synonomous variation had remained very limited. In some strains both orthologues and paralogues were present. It is concluded that CDC42 does not show any orthologous evolution, and that its paralogues haves the same function but are structurally relaxed. The variation or absence thereof could not be linked to ecological changes, from rock-inhabiting to pathogenic life style. It is concluded that eventual pathogenicity in Chaetothyriales is not expressed at the DNA level in CDC42 evolution. PMID- 19287535 TI - Coniosporium epidermidis sp. nov., a new species from human skin. AB - Coniosporium epidermidis sp. nov. is described from a superficial skin lesion with blackish discolouration in an 80-yr-old Chinese patient. The species produces dark, thick-walled, inflated, reluctantly liberating arthroconidia without longitudinal septa. Sequences of the ribosomal operon, as well as of the translation elongation factor 1-alpha support its novelty. The species is found in a lineage basal to the order Chaetothyriales, amidst relatives from rock, but also species repeatedly isolated from human skin and nails and eventually causing mild cutaneous infections. Coniosporium epidermidis is consistently found on humans, either asymptomatic or symptomatic. The species indicates a change of life style towards human pathogenicity, which is a recurrent type of ecology in derived Chaetothyriales. Superficial and cutaneous infection by melanized fungi is a new category in dermatology. PMID- 19287536 TI - Environmental isolation of black yeast-like fungi involved in human infection. AB - The present study focuses on potential agents of chromoblastomycosis and other endemic diseases in the state of Parana, Southern Brazil. Using a highly selective protocol for chaetothyrialean black yeasts and relatives, environmental samples from the living area of symptomatic patients were analysed. Additional strains were isolated from creosote-treated wood and hydrocarbon-polluted environments, as such polluted sites have been supposed to enhance black yeast prevalence. Isolates showed morphologies compatible with the traditional etiological agents of chromoblastomycosis, e.g. Fonsecaea pedrosoi and Phialophora verrucosa, and of agents of subcutaneous or systemic infections like Cladophialophora bantiana and Exophiala jeanselmei. Some agents of mild disease were indeed encountered. However, molecular analysis proved that most environmental strains differed from known etiologic agents of pronounced disease syndromes: they belonged to the same order, but mostly were undescribed species. Agents of chromoblastomycosis and systemic disease thus far are prevalent on the human host. The hydrocarbon-polluted environments yielded yet another spectrum of chaetothyrialean fungi. These observations are of great relevance because they allow us to distinguish between categories of opportunists, indicating possible differences in pathogenicity and virulence. PMID- 19287537 TI - The neurotropic black yeast Exophiala dermatitidis has a possible origin in the tropical rain forest. AB - The black yeast Exophiala dermatitidis is known as a rare etiologic agent of neurotropic infections in humans, occurring particularly in East and Southeast Asia. In search of its natural habitat, a large sampling was undertaken in temperate as well as in tropical climates. Sampling sites were selected on the basis of the origins of previously isolated strains, and on the basis of physiological properties of the species, which also determined a selective isolation protocol. The species was absent from outdoor environments in the temperate climate, but present at low abundance in comparable habitats in the tropics. Positive outdoor sites particularly included faeces of frugivorous birds and bats, in urban as well as in natural areas. Tropical fruits were found E. dermatitidis positive at low incidence. Of the human-made environments sampled, railway ties contaminated by human faeces and oily debris in the tropics were massively positive, while the known abundance of the fungus in steam baths was confirmed. On the basis of the species' oligotrophy, thermotolerance, acidotolerance, moderate osmotolerance, melanization and capsular yeast cells a natural life cycle in association with frugivorous animals in foci in the tropical rain forest, involving passage of living cells through the intestinal tract was hypothesized. The human-dominated environment may have become contaminated by ingestion of wild berries carrying fungal propagules. PMID- 19287538 TI - Selective factors involved in oil flotation isolation of black yeasts from the environment. AB - The oil flotation isolation technique has been successfully applied to recover chaetothyrialean black yeasts and relatives from the environment. The selective mechanisms playing a role in isolation are unknown. The fungi concerned are supposed to occupy specialized microniches in nature, taking advantage of (1) oligotrophism. Mineral oil as a main selective agent may be based on (2) hydrophobicity or on (3) assimilation. All three hypotheses are tested in this paper. Results show that cell wall hydrophobicity is unlikely to be a selective factor. Incubation under poor nutrient conditions provides competitive advantage for black yeasts, especially for Exophiala strains, which are subsequently enriched by mineral oil which enhances growth in this group of fungi. Incubation under mineral media and mineral oil can be used as selective factor. PMID- 19287539 TI - Characterisation of the substrate specificity of the nitrile hydrolyzing system of the acidotolerant black yeast Exophiala oligosperma R1. AB - The ;black yeast' Exophiala oligosperma R1 can utilise various organic nitriles under acidic conditions as nitrogen sources. The induction of a phenylacetonitrile converting activity was optimised by growing the strain in the presence of different nitriles and /or complex or inorganic nitrogen sources. The highest nitrile hydrolysing activity was observed with cells grown with 2 cyanopyridine and NaNO(3). The cells metabolised the inducer and grew with 2 cyanopyridine as sole source of nitrogen. Cell extracts converted various (substituted) benzonitriles and phenylacetonitriles. They usually converted the isomers carrying a substituent in the meta-position with higher relative activities than the corresponding para- or ortho-substituted isomers. Aliphatic substrates such as acrylonitrile and 2-hydroxy-3-butenenitrile were also hydrolysed. The highest specific activity was detected with 4-cyanopyridine. Most nitriles were almost exclusively converted to the corresponding acids and no or only low amounts of the corresponding amides were formed. The cells hydrolysed amides only with extremely low activities. It was therefore concluded that the cells harboured a nitrilase activity. The specific activities of whole cells and cell extracts were compared for different nitriles and evidence obtained for limitation in the substrate-uptake by whole cells. The conversion of 2-hydroxy-3 butenenitrile to 2-hydroxy-3-butenoic acid at pH 4 demonstrated the unique ability of cells of E. oligosperma R1 to hydrolyse aliphatic alpha hydroxynitriles under acidic conditions. The organism could grow with phenylacetonitrile as sole source of carbon, energy and nitrogen. The degradation of phenylacetonitrile presumably proceeds via phenylacetic acid, 2 hydroxyphenylacetic acid, 2,5-dihydroxyphenylacetic acid (homogentisate), maleylacetoacetate and fumarylacetoacetate. PMID- 19287540 TI - Biodiversity of the genus Cladophialophora. AB - Cladophialophora is a genus of black yeast-like fungi comprising a number of clinically highly significant species in addition to environmental taxa. The genus has previously been characterized by branched chains of ellipsoidal to fusiform conidia. However, this character was shown to have evolved several times independently in the order Chaetothyriales. On the basis of a multigene phylogeny (nucLSU, nucSSU, RPB1), most of the species of Cladophialophora (including its generic type C. carrionii) belong to a monophyletic group comprising two main clades (carrionii- and bantiana-clades). The genus includes species causing chromoblastomycosis and other skin infections, as well as disseminated and cerebral infections, often in immunocompetent individuals. In the present study, multilocus phylogenetic analyses were combined to a morphological study to characterize phenetically similar Cladophialophora strains. Sequences of the ITS region, partial Translation Elongation Factor 1-alpha and beta-Tubulin genes were analysed for a set of 48 strains. Four novel species were discovered, originating from soft drinks, alkylbenzene-polluted soil, and infected patients. Membership of the both carrionii and bantiana clades might be indicative of potential virulence to humans. PMID- 19287542 TI - Is quality affordable? PMID- 19287541 TI - Leaf-inhabiting genera of the Gnomoniaceae, Diaporthales. AB - The Gnomoniaceae are characterised by ascomata that are generally immersed, solitary, without a stroma, or aggregated with a rudimentary stroma, in herbaceous plant material especially in leaves, twigs or stems, but also in bark or wood. The ascomata are black, soft-textured, thin-walled, and pseudoparenchymatous with one or more central or eccentric necks. The asci usually have a distinct apical ring. The Gnomoniaceae includes species having ascospores that are small, mostly less than 25 mum long, although some are longer, and range in septation from non-septate to one-septate, rarely multi septate. Molecular studies of the Gnomoniaceae suggest that the traditional classification of genera based on characteristics of the ascomata such as position of the neck and ascospores such as septation have resulted in genera that are not monophyletic. In this paper the concepts of the leaf-inhabiting genera in the Gnomoniaceae are reevaluated using multiple genes, specifically nrLSU, translation elongation factor 1-alpha (tef1-alpha), and RNA polymerase II second largest subunit (rpb2) for 64 isolates. ITS sequences were generated for 322 isolates. Six genera of leaf-inhabiting Gnomoniaceae are defined based on placement of their type species within the multigene phylogeny. The new monotypic genus Ambarignomonia is established for an unusual species, A. petiolorum. A key to 59 species of leaf-inhabiting Gnomoniaceae is presented and 22 species of Gnomoniaceae are described and illustrated. PMID- 19287543 TI - Improving the patient's experience. PMID- 19287544 TI - Improving surgical outcomes. PMID- 19287545 TI - Case study: improving the management of eye care programmes. PMID- 19287547 TI - Community Eye Health MSc dissertations. PMID- 19287546 TI - Short-term visits by eye care professionals: ensuring greater benefit to the host community. PMID- 19287548 TI - A brief experience of blindness. PMID- 19287549 TI - Epilation of eyelashes. PMID- 19287550 TI - Patient follow-up in primary care after behavioral health screening in an urban public hospital system: a prospective, observational study of 2686 patients. AB - OBJECTIVE: Patients with psychiatric conditions are known to experience poor and often disparate health outcomes. To investigate one potential mechanism for this phenomenon, we examined whether patients who screen positive for psychiatric comorbidity are lost to follow-up from primary care at higher rates than screen negative controls. METHOD: Patients in a public hospital system were followed prospectively for an 18-month period after an initial routine behavioral health screening in neighborhood health centers. Screening data were linked to electronic medical record visit data, and loss to follow-up was ascertained using Cox proportional hazards modeling. RESULTS: A public hospital health program screened 2686 patients from March 1998 to December 2000, and their visits were counted prospectively for 18 months. Nearly one third (N = 772, 29%) screened positive for a psychiatric condition. The screen-positive group had lower rates of censoring and a shorter time-to-event than the controls, indicating a higher continuing visit rate in primary care. This relationship persisted after adjustment for demographic variables, insurance type, substance abuse, and violence exposure. CONCLUSION: Patients who screen positive for psychiatric comorbidity are not lost to follow-up at higher rates than screen-negative controls. This finding suggests that disparate outcomes for mentally ill patients in a public hospital system may not be based on reduced access to or lack of contact with primary care providers. Further study of systems or provider-related factors is needed to ascertain the pathways toward poor health for this population. PMID- 19287551 TI - Lithium and psoriasis: what primary care and family physicians should know. AB - OBJECTIVE: This review focuses on the association of lithium treatment and psoriasis. The mechanism of action of lithium in causing psoriasis and the clinical presentation of psoriasis secondary to lithium treatment are considered. DATA SOURCES: A search of the literature from 1949 to 2007 was performed using MEDLINE, with the following search terms: lithium, psoriasis, skin, dermatology, and psychodermatology. DATA SYNTHESIS: Lithium is involved in a variety of cutaneous reactions including psoriasis, which may present as exacerbation of preexisting psoriasis, induction of de novo psoriasis, pustular psoriasis, nail changes, and psoriatic arthropathy. The appearance of psoriatic lesions may occur at normal therapeutic serum lithium levels. The refractory period for the development of psoriatic lesions is variable and generally longer in induction and shorter in exacerbation of psoriasis. Lithium-induced psoriasis is often resistant to conventional treatment modalities, and some cases may require dose reduction or discontinuation of lithium treatment. CONCLUSION: Lithium is the mainstay of treatment in bipolar disorder and is associated with a variety of cutaneous side effects including psoriasis. Primary care providers and family physicians should be knowledgeable about the association of lithium and its dermatologic side effects. Early recognition and management could be beneficial in avoiding the issues of noncompliance and further deterioration of mood symptoms secondary to obviously disfiguring skin appearance. Primary care, psychiatry, and dermatology liaison services will prove helpful in managing these patients. PMID- 19287552 TI - Examining the efficacy of adjunctive aripiprazole in major depressive disorder: a pooled analysis of 2 studies. AB - BACKGROUND: Patients with major depressive disorder (MDD) who fail to achieve complete remission with antidepressant therapy may benefit from augmentation therapy with an atypical antipsychotic. METHOD: A pooled analysis was performed on 2 identical 14-week studies (8-week prospective antidepressant therapy treatment phase followed by 6-week randomized double-blind phase) evaluating the efficacy of adjunctive aripiprazole (2-20 mg/day) in DSM-IV-TR-defined MDD patients with an inadequate response to antidepressant therapy. Primary efficacy endpoint was the mean change in Montgomery-Asberg Depression Rating Scale (MADRS) total score from end of the prospective phase (week 8) to end of randomized phase (week 14, last observation carried forward). Subgroup analyses were performed. The key secondary endpoint was mean change in Sheehan Disability Scale (SDS) mean score. RESULTS: At endpoint, mean change in MADRS total score was significantly greater with adjunctive aripiprazole (-8.7) than with adjunctive placebo (-5.7; p < .001). Except for a differential treatment-by-sex interaction, change in MADRS total scores were consistently greater with adjunctive aripiprazole than with adjunctive placebo, regardless of race, age, episode duration, prior antidepressant therapy response, number of historical treatment failures, severity of depressive symptoms, and antidepressant. At endpoint, MADRS remission rates were significantly greater with adjunctive aripiprazole than with placebo (25.7% vs. 15.4%; p < .001). Adjunctive aripiprazole also demonstrated significantly greater improvements in mean change from baseline in SDS total score than adjunctive placebo (-1.2 vs. -0.6; p = .001). CONCLUSION: Augmentation of antidepressant therapy with the atypical antipsychotic aripiprazole resulted in significant efficacy benefits across a range of subgroups of patients with MDD. Further study of a treatment-by-sex interaction is needed. TRIAL REGISTRATION: www.clinicaltrials.gov Identifiers: NCT00095823 and NCT00095758. PMID- 19287553 TI - The relationship between medically self-sabotaging behaviors and borderline personality disorder among psychiatric inpatients. AB - OBJECTIVE: In this study, we hypothesized and explored a relationship between medically self-sabotaging behaviors and borderline personality disorder. METHOD: Using a cross-sectional self-report survey methodology, we examined 120 psychiatric inpatients, who were not psychotic, demented, medically ill, or cognitively impaired, being treated in an urban community hospital located in a midsized, midwestern city (sample of convenience) for medically self-sabotaging behaviors (author-developed survey) and borderline personality disorder. Borderline personality disorder was assessed with the following 3 measures: the borderline personality scale of the Personality Diagnostic Questionnaire-4 (PDQ 4), the Self-Harm Inventory (SHI), and the McLean Screening Inventory for Borderline Personality Disorder (MSI-BPD). Data were collected from May 2006 to November 2007. RESULTS: For the 76 respondents (63.3%) who reported having engaged in at least 1 medically self-sabotaging behavior, the mean number of different medically self-sabotaging behaviors was 4.11 (SD = 3.93). With regard to the most commonly endorsed behaviors, approximately one quarter of participants acknowledged damaging self on purpose and seeking medical treatment; not going for medical treatment, despite needing it, to purposefully hurt self; not taking a prescribed medication to hurt self; and gravitating toward a dangerous situation hoping to be physically hurt. As hypothesized, greater numbers of self-reported medically self-sabotaging behaviors were related to higher scores on the PDQ-4 (r = 0.28, p < .01), the SHI (r = 0.55, p < .001), and the MSI-BPD (r = 0.41, p < .001). CONCLUSIONS: Medically self-sabotaging behaviors are commonly encountered in psychiatric inpatients with borderline personality disorder. PMID- 19287554 TI - Symptoms of posttraumatic stress disorder after ritual female genital surgery among bedouin in Israel: myth or reality? AB - OBJECTIVE: Ritual female genital surgery (RFGS), or female circumcision, is common among certain ethnic groups in Asia and Africa and describes a range of practices involving complete or partial removal of the female external genitalia for nonmedical reasons. Several studies in African populations, in which more severe forms of RFGS are performed, reported an increased prevalence of posttraumatic stress disorder and other psychiatric syndromes among circumcised women than among uncircumcised controls. Among the Bedouin population in southern Israel, RFGS has become a symbolic operation without major mutilation. However, in a study performed in 1999, Bedouin women after RFGS reported difficulties in mother-daughter relationships and trust. This pilot study assessed the mental health of Bedouin women from southern Israel after RFGS compared to age-matched controls without RFGS. METHOD: The psychological impact of RFGS was assessed in 19 circumcised Bedouin women compared to 18 age-matched controls. The Post Traumatic Stress Disorder Scale, Symptom Checklist, Impact of Event Scale, and a demographics and background questionnaire were used to assess traumatization and psychiatric illnesses. The study was conducted from March to July 2007. RESULTS: No statistically significant differences were found between the 2 groups. CONCLUSIONS: The prevailing procedure of RFGS among the Bedouin population of southern Israel had no apparent effect on mental health. PMID- 19287555 TI - Immediate and 8-month impact of a medical educational course for general practitioners on knowledge about schizophrenia and its treatment: results of a 3 phase study from Brescia, Italy. AB - OBJECTIVE: To test the efficacy of a training course on the diagnosis and treatment of schizophrenia, tailored for the general practitioner. METHOD: A course, in a 3-session format, was given to 215 primary care doctors from the city of Brescia and its province, in Italy. All 706 doctors working in primary care were asked to participate. Of these doctors, 30.5% took part in the study. The first session (215 doctors) assessed baseline knowledge of schizophrenia (June 2002), the second (173 doctors) gave formal teaching and assessed post lesson knowledge (October 2002), and the third (130 doctors) evaluated the retention of knowledge after 8 months (July 2003). The main outcome measures were total number of schizophrenia symptoms identified, total number of antipsychotics identified, and knowledge about antipsychotic-related adverse events. RESULTS: Post-lesson, general practitioners could identify 6.5 more symptoms (p < .001) and 4.9 more antipsychotics (p < .001). Compared to baseline, 71.5% vs. 15.4% of doctors had a good knowledge of antipsychotic-related adverse events. Although a loss of knowledge was found after the 8-month follow-up, knowledge at the endpoint was significantly higher than at baseline for the 3 main outcome variables (p < .001). CONCLUSION: The teaching course on schizophrenia for general practitioners was effective, and the knowledge gained after teaching was stable across time. PMID- 19287556 TI - Prevalence of depressive symptoms and associated factors in elderly primary care patients: a descriptive study. AB - BACKGROUND: Depressive symptoms are common in older adults. A majority will be seen in primary care. The aim was to study the prevalence of and to explore factors associated with depressive symptoms in elderly primary care patients. METHOD: In consecutive patients aged 60 years and older attending a Swedish primary care center between February and December of 2003, depressive symptoms were identified as >= 13 points on the Montgomery-Asberg Depression Rating Scale Self-Rated version (MADRS-S). Somatic symptoms measured according to PRIME-MD, age, socioeconomic status, gender, somatic diagnoses, and medication were analyzed in relation to presence of depressive symptoms. RESULTS: Forty-six of 302 patients (15%) rated themselves in the depressed range. There were no differences between depressed and nondepressed patients concerning socioeconomic status, other illnesses, or medication except for use of sedatives/hypnotics being more common (OR = 2.7, 95% CI = 1.3 to 5.6) in depressed patients. Patients in the group scoring >= 13 on the MADRS-S were more likely to have become widowed during the last year (OR = 6.0, 95% CI = 1.7 to 20.8) or to have indicated significant life events (OR = 4.3, 95% CI = 2.0 to 9.0), but were less likely to report having leisure time activities (OR = 0.2, 95% CI = 0.08 to 0.41) or perception of good health (OR = 0.1, 95% CI = 0.05 to 0.3). Patients being treated for depression did not have increased depression scores (OR = 1.4, 95% CI = 0.66 to 3.1). CONCLUSION: In a group of unselected primary care elderly patients, the prevalence of depressive symptoms was high. Use of sedatives/hypnotics was remarkably common in patients with depressive symptoms. Patients with ongoing treatment of depression did not have increased depression scores, indicating the good prognosis for treated depression in the elderly. PMID- 19287557 TI - Adult ADHD pharmacotherapy. PMID- 19287558 TI - Methadone-Associated QTc Prolongation: A Case Report and Review of the Literature. PMID- 19287559 TI - Casebook of a cancer therapist, 1. PMID- 19287560 TI - Mood follows the market. PMID- 19287561 TI - Aripiprazole in schizotypal personality disorder: a case report. PMID- 19287562 TI - Risperidone- and aripiprazole-induced leukopenia: a case report. PMID- 19287563 TI - Vigilance needed in recognizing behavior changes in patients taking statins. PMID- 19287564 TI - Aggressive behavior after ingestion of a high dose of sildenafil. PMID- 19287565 TI - A case of pancreatitis associated with aripiprazole in the absence of hyperglycemia. PMID- 19287566 TI - CALM: A Mnemonic for Treatment Options for Bipolar Disorder. PMID- 19287567 TI - Suicide attempt and psychosis revealed after an apparent traffic accident: a case report. PMID- 19287569 TI - Erratum. AB - [This corrects the article on p. 72 in vol. 10, PMID: 18311428.]. PMID- 19287568 TI - Metabolic side effects of risperidone in children and adolescents with early onset schizophrenia. PMID- 19287570 TI - Effect of Copper Sulfate and Lead Acetate on Infection of Pines with Bursaphelenchus xylophilus. AB - Treatment of 3-year-old Scots, white, and Austrian pine seedlings with copper sulfate or lead acetate significantly affected energy homeostasis and oleoresin production in the seedlings but did not induce wilting of the seedlings. Inoculation of copper sulfate-treated or lead acetate-treated white, Scots, and Austrian pine seedlings with the white pine specific pathotype of Bursaphelenchus xylophilus, VPSt-1, caused a significant increase in oleoresin production, stressed energy homeostasis, and induced rapid wilting of the seedlings. Scots pine lost tolerance and Austrian pine lost resistance to VPSt-1 after the seedlings were treated with either copper sulfate or lead acetate. These results suggest that environmental pollution may significantly affect susceptibility of pines to B. xylophilus and may have a role in establishment of this nematode in uninfested areas. PMID- 19287571 TI - Ditylenchus destructor in Hulls and Seeds of Peanut. AB - The potato rot nematode, Ditylenchus destructor Thorne, is reported for the first time in hulls and seeds of peanut. The populations found differed from D. dipsaei and D. myceliophagus in habitat, number of lateral incisures, shape of tail tip, and length of postvulval sac. Infected hulls had brown necrotic tissue at the point of connection with the peg, and a black discoloration appeared first along the longitudinal veins. Infected seeds were usually shrunken, and testae and embryos had a yellow to brown or black discoloration. Of 877 seed samples graded "damaged" from all major peanut producing areas of South Africa, 73% were infected. PMID- 19287572 TI - Control of Globodera tabacum solanacearum by Alternating Host Resistance and Nematicide. AB - The feasibility of alternating use of resistant vs. susceptible flue-cured tobacco cultivars to improve control of Globodera tabacum subsp, solanacearum (TCN) was investigated at two Virginia locations in 1984-86. Post-harvest TCN population densities were reduced in each year of the study when fenamiphos was used with a TCN-resistant cultivar (NC 567), relative to susceptible cultivars (K 326 or Mc 944). Using NC 567 with fenamipbos also reduced preplant TCN population densities in the next growing season. Egg population densities before planting in 1986 were significantly lower in plots planted with NC 567 in 1984, even when a susceptible cultivar had been planted in 1985. Use of fenamiphos with NC 567 in 1984 and 1985 further reduced preplant egg population densities in 1986. Economic returns were significantly greater in 1984 when NC 567 was used with fenamiphos, rather than a susceptible cultivar. Treatments involving fenamiphos and (or) NC 567 in 1984 and 1985 resulted in higher economic returns in 1986 than did treatments using a susceptible cultivar without fenamiphos in both previous years. Economic returns were highest in 1986 when fenamiphos and NC 567 were used in 1984 and 1985 and a susceptible cultivar was planted in 1986. PMID- 19287573 TI - Development of Heterodera glycines as Affected by Alachlor and Fenamiphos. AB - A series of greenhouse experiments was conducted to elucidate the postinfection development of Heterodera glycines in response to applications of alachlor and fenamiphos. The rate of H. glycines maturation on a susceptible soybean cultivar was not altered by 1.0 mug alachlor/g soil but was completely inhibited by 1.0 or 1.5 mug fenamiphos/g soil. An alachlor-fenamiphos combination allowed development after an initial 300-degree-day delay. Nematode maturation on the resistant soybean cultivar Centennial with 1.0 mug alachlor/g soil was similar to that observed on an untreated resistant control. Twice as many females matured on Centennial plants growing in alachlor-treated soil as on untreated Centennial plants. Fenamiphos in combination with alachlor (1.0 mug a.i./g soil) allowed development on Centennial at half the rate of the resistant control. This antagonism between alachlor and fenamiphos on development may help to explain late season population resurgence of H. glycines observed with field application of these pesticides. PMID- 19287574 TI - Effect of Planting Date, Alachlor, and Fenamiphos on Heterodera glycines Development. AB - The effects of alachlor (2.25 kg a.i./ha) and fenamiphos (2.25 kg a.i./ha) on the penetration and development of Heterodera glycines were examined on Glycine max cultivars Deltapine 105 planted 29 April, 29 May, and 29 June 1986 and Deltapine 105 and Centennial planted 15 May, 15 June, and 15 July 1987. Penetration was lowest on the third planting of soybeans and on fenamiphos-treated plants. Development from second-stage juveniles to adult females required 270 (1986) and 260 (1987) DD20/32 on roots from the first planting control and alachlor treatments. Fenamiphos, alone or with alachlor, retarded development in Deltapine 105 (1986) and in Centennial (1987). Males matured in roots from the second planting in 190 (1986) and 180 (1987) DD20/32 regardless of treatment or cultivar. No development occurred in roots from the third planting until 400 DD20/32 in 1986, but in 1987 development was similar to that in roots from the second planting. Nematode development was similar in alachlor-treated and control roots regardless of planting date. Fenamiphos restricted nematode penetration on most planting dates and slowed development. Simultaneous applications of alachlor and fenamiphos usually also inhibited development. PMID- 19287575 TI - Resistance to Meloidogyne spp. in Allohexaploid Wheat Derived from Triticum turgidum and Aegilops squarrosa. AB - Expression of resistance to Meloidogyne incognita and M. javanica from Aegilops squarrosa was studied in a synthetic allohexaploid produced from Triticum turgidum var. durum cv. Produra and Ae. squarrosa G 3489. The reproductive rate of different races of M. incognita and M. javanica, expressed in eggs per gram of fresh root, was low (P < 0.05) on the synthetic allohexaploid and the resistant parent, Ae. squarrosa G 3489, compared with different bread and durum wheat cultivars. Reproduction of race 2 and race 3 of M. incognita and an isolate of M. javanica was studied on the synthetic allohexaploid and seven cultivars of T. aestivum: Anza, Coker 747, Coker 68-15, Delta Queen, Double Crop, McNair 1813, and Southern Bell. The latter six cultivars are grown in the southeastern United States and reportedly were resistant to M. incognita. Significant differences (P < 0.05) were detected in nematode reproduction on the seven bread wheat cultivars. Reproduction of M. incognita race 3 and M. javanica was highest on Anza. Reproductive rates on the six southeastern United States bread wheat cultivars varied both within and among nematode isolates. The lowest reproductive rates of the three root-knot isolates were detected in the synthetic allohexaploid. PMID- 19287576 TI - Four New Species of Heteroderidae Including Ekphymatodera n. gen. from California. AB - Four new species and a new genus of Heteroderidae from California are described, and their significance for phylogenetic analysis of the family is discussed. The new genus with type species, Ekphymatodera thomasoni n. gen., n. sp., shares many characteristics with Hylonema Luc, Taylor, &Cadet, 1978, but it is distinguished by its much greater vulva-anus distance and unique cuticular pattern. Hypotheses of relationships of Ekphymatodera and Hylonema with Sarisodera Wouts and Sher, 1971 versus Heterodera Schmidt, 1871 are discussed. Verutus californicus n. sp. is larger than the type species, Verutus volvingentis Esser, 1981, differing in females particularly by the greater distance of its excretory pore from the anterior end. Monophyly of Verutus, which may be an outgroup of all other Heteroderidae, is strengthened by description of V. californicus. Atalodera trilineata n. sp. differs from other ataloderines by having second-stage juveniles with three lateral lines and from the type, Atalodera ucri Wouts and Sher, 1971, by the more subtle cuticular pattern of females and longer juveniles with much longer tails. Atalodera festucae n. sp., with four lateral lines in juveniles, has smaller females than A. trilineata and has a protruding dorsal vulval lip. A unique common ancestor for Atalodera-Sherodera-Thecavermiculatus is supported, and monophyly with Thecavermieulatus andinus Golden, Franco, Jatala, &Astocaza, 1973 is considered. PMID- 19287577 TI - Influence of Glomus intraradices and Soil Phosphorus on Meloidogyne incognita Infecting Cucumis melo. AB - The interaction among Glomus intraradices, Meloidogyne incognita, and cantaloupe was studied at three soil phosphorus (P) levels in a greenhouse. All plants grew poorly in soil not amended with P, regardless of mycorrhizal or nematode status. In soil amended with 50 mug P /g soil, M. incognita suppressed the growth of nonmycorrhizal plants by 84%. In contrast, growth of mycorrhizal plants inoculated with M. incognita was retarded by only 21%. A similar trend occurred in plants grown in soil with 100 mug P /g soil. Mycorrhizal infection had no effect on the degree of root-knot gall formation and did not affect the number of nematode eggs per egg mass. Mineral levels in plant shoots generally declined as soil P levels increased and were not significantly influenced by G. intraradices or M. incognita. PMID- 19287578 TI - Impact of Meloidogyne incognita on Physiological Efficiency of Vitis vinifera. AB - Four-week-old French Colombard plants rooted from green cuttings were inoculated with 0, 1,000, 2,000, 4,000, or 8,000 Meloidogyne incognita second-stage juveniles and maintained at 25 C night and 30 C day. Leaf area and dry weight and the rates of photosynthesis, stomatal conductance, and internal leaf CO concentration were measured at intervals up to 59 days after inoculation. Nematode stress dosage, measured as the product of cumulative number of juveniles and females and their total energy (calories) demand, was up to 3.4 kcal and accounted for up to 15% of the energy assimilated by the plants. There was a decline in the rate of leaf area expansion and leaf, stem, shoot, root (excluding nematode weight), and total plant dry weight with increasing nematode stress. Root weight including nematodes was not affected. Total respiration, plant photosynthesis, energy assimilated into plant tissue and respiration, and gross production efficiency decreased significantly with nematode stress. Photosynthetic rate, transpiration rate, stomatal conductance, and internal CO concentration were not affected. This study demonstrates that the energy demand for growth and reproduction of M. incognita accounts for a significant portion of the total energy entering the plant system. As a result, less energy is partitioned into leaf area expansion which, in turn, affects the energy entering the system and results in decreased productivity of nematode-infected grape vines. PMID- 19287579 TI - Polyamine Synthesis by the Mermithid Nematode Romanomermis culicivorax. AB - The polyamine and amino acid composition of the mermithid nematode, Romanomermis culicivorax, and its host, Aedes aegypti, was determined. Putrescine, spermidine, spermine, cadaverine and two acetylated spermidine derivatives were present in parasitic juveniles, newly-emerged post-parasites, and eggs of R. culicivorax. Whole insect homogenates of fourth-instar A. aegypti contained the same array of polyamines, except that the putrescine:spermidine ratio was the inverse of that in parasitic R. culicivorax. Polyamines and amino acids were in greater concentrations in the nematode eggs than in other developmental stages investigated. Both the host and nematode possess the biosynthetic capacity for polyamine biosynthesis, as evidenced by measurable activities of ornithine decarboxylase in the host's tissues and the nematode's free-living stages. PMID- 19287580 TI - Pathogenicity of Two Populations of Meloidogyne hapla Chitwood on Alfalfa and Sainfoin. AB - The pathogenicity of two populations of the northern root-knot nematode, Meloidogyne hapla Chitwood, population 1 (P1) from alfalfa and population 2 (P2) from sainfoin, was studied on both alfalfa and sainfoin for 25 weeks. Alfalfa and sainfoin plants inoculated with P2 had significantly (P 10 C (DD10). Hatch occurred below the reported 18 C activity threshold, was restricted below 12 C, and was inhibited below 10 C. Soil moisture influence on hatch was measured by placing egg masses in Hesperia sandy loam and subjecting them to suction pressures ranging from -1.1 bars to -4 .5 bars. Suction potentials of less than -2 bars reduced hatch and less than -3 bars inhibited hatch. J2 were placed in sandy loam soil with soil moisture near field capacity, and their motility was measured over a period of 500 DD10. In the absence of a host, more than 90% of J2 became nonmotile over this period. PMID- 19287617 TI - Description of Discocriconemella mineira n. sp. and D. degrissei Males from Brazil. AB - A new species of Discocriconemella and the male of Discocriconemella degrissei are described from Minas Gerais state of Brazil. Discocriconemella mineira n. sp. differs from D. discolabia by having a longer stylet (61-71 vs. 35-47 mum) and fewer body annuli (78-88 vs. 155-174), and from D. inarata by the longer stylet (61-71 vs. 51-61 'm) and shorter body length (253-342 vs. 354-486 mum). It differs from D. persae by having a shorter stylet, fewer body annuli, and a shorter post vulval body portion (Rv = 6-7 vs. 14-20). Discocriconemella mineira can be separated from D. degrissei by the head disc which is round in D. mineira and quadrilobate in D. degrissei. Males of D. degrissei resemble those of D. retroversa. PMID- 19287618 TI - Esterase and malate dehydrogenase phenotypes in portuguese populations of meloidogyne species. AB - Nonspecific esterases and malate dehydrogenases of 1-5 females from 40 root-knot nematode populations from Portugal were analyzed by electrophoresis in 0.4-mm thick polyacrylamide gels. Fourteen major bands of esterase activity were detected, corresponding to 10 distinct phenotypes, Meloidogyne javanica and M. hapla had distinct species-specific phenotypes. Two phenotypes occurred in M. arenaria. The most variability was found among M. incognita populations. Of the remaining two phenotypes, one was associated with M. hispanica and the other belonged to a new species. Three malate dehydrogenase phenotypes were discerned on the basis of particular combinations of the eight main bands of activity found. As previously found, esterases were more useful than malate dehydrogenases in identification of the major Meloidogyne species. The host plant had no effect on the nematode esterase or malate dehydrogenase phenotypes. PMID- 19287619 TI - Survival of Heterodera zeae in Soil in the Field and in the Laboratory. AB - Eggs and (or) second-stage juveniles (J2) inside cysts of Heterodera zeae survived over winter in the field with no detectable mortality at all six depths to 30 cm from which soil samples were collected between corn stubble in the row at 4-8-week intervals. Few or no free J2 were recovered from soil collected in January-April from the top 5 cm, but some were recovered at all samplings from soil collected at greater depths. Emergence of J2 from cysts and numbers of females developing on corn roots in bioassays of cysts increased substantially between January and April. Cyst numbers in a fallow area of the corn field did not decline at any depth to 30 cm during 20 months. Free soil J2, J2 emerged from cysts, and females from the bioassay of cysts were highest at the first soil sampling in July after 10 months of fallow; numbers of nematodes in all three categories declined thereafter, but a few were still detectable after 20 months of fallow. Some cysts were still being recovered after 51 months from naturally infested field soil stored moist in the laboratory at 2 C and 24 C. Females were produced in the bioassays of cysts recovered from soil stored for 38 months at 24 C and for 32 months at 2 C. No free J2 were recovered from soil after 1 month of storage at -18 C, but even after 7 months storage J2 emerged from cysts that were recovered and many females developed in bioassays of those cysts. PMID- 19287620 TI - Endoparasitic nematodes in maize roots in the Western transvaal as related to soil texture and rainfall. AB - Eight endoparasitic nematode species were recovered from 170 maize root samples in western Transvaal, Republic of South Africa. Pratylenchus zeae had the highest average population density (17,454/5 g roots), followed by P. neglectus (5,827/5 g roots), P. penetrans (5,617/5 g roots), P. brachyurus (3,060/5 g roots), Meloidogyne incognita plus M. javanica (301 juveniles/5 g roots), P. crenatus (130/5 g roots), and Rotylenchutus parvus (64/5 g roots). The 17 reasonably homogeneous farming areas (RHFA) surveyed could be ranked on the basis of the incidence of the prevalent nematode species. A positive relationship was found between the incidence of P. brachyurus and R. parvus and long-term average annual rainfall. The incidence of P. penetrans and the Meloidogyne spp. was positively related to a combination of sand percentage and long-term average annual rainfall. PMID- 19287621 TI - Influence of Temperature and Soybean Phenology on Dormancy Induction of Heterodera glycines. AB - Growth room and field experiments were conducted to determine the influence of soil temperature and soybean phenology on dormancy induction of a North Carolina population of Heterodera glycines race 1. Three temperature regimes and two photoperiods to regulate plant phenology were investigated in growth rooms. H. glycines hatch was greatest from the 26 and 22 C (day and night) temperature treatment, intermediate at 22 and 18 C, and least from the decreasing regime (26 and 22 C, 22 and 18 C, and 18 and 14 C). More eggs hatched and greater nematode reproduction occurred on pod-producing soybeans than on those that remained vegetative. In the field study, hatching patterns were not different between depodded and naturally senescing soybeans nor between the different maturity groups of soybean cultivars (groups V through VIII). Egg hatch (9-16%) was greatest in August and September when mean soil temperatures were between 25 and 29 C. Hatch declined to 1% in vitro and was not detectable in the bioassay in November. Greatest nematode numbers were observed on the latest maturing cultivar (group VIII) and fewest on the cultivar which matured earliest (group V). Decreasing temperature appears to be more important than soybean phenology in dormancy induction of H. glycines. PMID- 19287622 TI - Accelerated movement of nematodes from soil in baermann funnels with temperature gradients. AB - Baermann funnels were modified to eliminate or reverse the small temperature gradient (1-2 C/cm) across the soil layer that normally results from water evaporation. Effects of modifications on extraction efficiency were examined at various ambient temperatures and after overnight adaptation of three nematode species at 20 and 30 C. Extraction of Meloidogyne incognita from sandy loam, Tylenchulus semipenetrans from sandy clay loam, and Rotylenchulus reniformis from silt was greatly accelerated simply by covering funnels to prevent evaporation. In most cases, covering increased the nematodes extracted by 10-100 times after 5.5-48 hours. Faster and more efficient extraction of R. reniformis occurred over a wide range of ambient temperature (18-29 C). Effects of ambient temperature and temperature gradient direction on Baermann funnel extraction of R. reniformis were partly inconsistent with the behavior of R. reniformis in agar. Nematodes in agar moved toward cold at some ambient temperatures and toward heat at other temperatures. They always appeared to move toward cold on Baermann funnels. Differences were not attributable to blockage of gas exchange by covers. In agar and in funnels, the patterns of response to ambient temperature were shifted in the direction of the storage temperature. PMID- 19287623 TI - Screening Subterranean Clover (Trifolium spp.) Germplasm for Resistance to Meloidogyne species. AB - This study was conducted to identify lines of subterranean clover (Trifolium spp.) with resistance to Meloidogyne arenaria (Neal, 1989) Chitwood, 1949, race 1; M. incognita (Kofoid and White, 1919) Chitwood, 1949, race 3; and M. javanica (Treub, 1885) Chitwood, 1949. A collection of 134 subterranean clover lines was evaluated and all had intermediate to high susceptibility. Root galling was negatively correlated with both seed and dry matter yields. Soil fumigation significantly reduced the nematode population in the field. Results indicate there is limited genetic resistance to root-knot nematodes among subterranean clover lines. Alternative sources of variation for this trait should be investigated. PMID- 19287624 TI - Cereal Cyst Nematode (Heterodera avenae) on Oats. II. Early Root Development and Nematode Tolerance. AB - The effect of Heterodera avenae infestation on early seminal and lateral root growth was examined in four oat genotypes differing in tolerance to H. avenae. Recently emerged seminal roots were inoculated with a range of H. avenae larval densities, then transferred a hydroponic system to remove the effect of later nematode penetration on root development. Intolerance to H. avenae was assessed in terms of impairment of seminal root extension resulting in fewer primary lateral roots emerging from the seminal root below the zone of juvenile penetration. Tolerant plants infested with H. avenae had longer lateral root systems than infested intolerant plants. The decline in lateral root growth below the penetration zone was partly offset by increased growth above. This did not contribute to tolerance, however, as there were no differences between cultivars for this feature. Nematodes induced earlier nodal root emergence in all cultivars. Nodal root development was most advanced on the most tolerant cultivar. PMID- 19287625 TI - Monoclonal antibodies to secretory granules in esophageal glands of meloidogyne species. AB - Monoclonal antibodies to secretory granules in the dorsal or subventral esophageal glands were generated by injecting BALB/c mice with immunogens from preparasitic second-stage juveniles (J2) of Meloidogyne incognita. Antibodies specific for secretory granules in the J2 subventral esophageal glands or the dorsal gland were identified by indirect immunofluorescence microscopy. Only antibodies that reacted with granules in the J2 dorsal gland reacted with the esophageal gland lobe ofM. incognita adult females. The antibodies also reacted with secretory granules in both types of esophageal glands in M. javanica and M. arenaria J2 but not with granules in esophageal glands of Heterodera glycines J2. PMID- 19287626 TI - Nematode autofluorescence and its use as an indicator of viability. AB - Representatives of 15 nematode genera were viewed with 450-490-nm epi illumination and found to autofluoresce. The autofluorescence was limited to 1-5 mum-d globules in the intestinal cells of live nematodes. When adult Pratylenchus penetrans or Caenorhabditis elegans were killed with formaldehyde, freezing, or heat, autofluorescence dispersed throughout the body. Mixed stages of P. penetrans were killed by freezing at several different temperatures. Estimates of survival based on autofluorescence dispersal matched estimates based on mobility more closely than did estimates based on the vital stain, eosin-y. PMID- 19287627 TI - Abundance and Vertical Distribution of Longidorus breviannulatus Associated with Corn and Potato. AB - Longidorus breviannulatus was detected in a field planted to corn after 13 years of potato. Nematode populations were maintained in this field in adjacent corn and potato plots for 2 years but did not increase significantly on either crop. Population levels increased until approximately 60 days after planting and then declined until the end of the growing season. Overwinter mortality was negligible. The vertical distribution of the nematode population changed during the course of the season. More nematodes were recovered from depths of 0-15 cm in early season samples and from depths of 15-30 cm in late season samples. Data indicated that this redistribution was due to nematode migration. PMID- 19287628 TI - Distribution of Pratylenchus scribneri between Root and Soil Habitats. AB - The abundance of Pratylenchus scribneri in soil and root habitats was compared in potato and corn plots during 1986-88. Nematodes were extracted from 100-cm(3) soil samples and the roots contained within the samples. The percentage of the population recovered from soil, similar among years and crops, averaged ca. 50% at the beginning and end of the growing season and ca. 20% from early to late season. Proportionately more adults and fourth-stage juveniles than younger stages were located outside roots until harvest. In a related study, nematodes were isolated from the roots, root surfaces, and soil associated with roots of whole corn and potato plants sampled from the field. Nematode population estimates calculated from the whole plant samples were generally lower than those based on soil cores, but showed similar patterns of population growth. Nematode density per gram dry weight was highest in roots, intermediate on root surfaces, and lowest in soil. Estimates of the absolute abundance of nematodes in each of the three habitats were highest in roots or soil, depending on the sampling date, and lowest on root surfaces. This study demonstrates that P. scribneri inhabits soil environments even when host roots are present and illustrates the importance of considering all possible habitats when estimating the size of Pratylenchus spp. populations. PMID- 19287629 TI - Effect of Temperature on Development and Survival of the Mermithid Filipjevimermis leipsandra. AB - The development and survival of egg, juvenile (parasitic), preovipositing, and ovipositing stages of the parasitic mermithid Filipjevimermis leipsandra were determined at eight temperatures. Infection of the host, Diabrotica balteata, peaked at 30 C and fell off sharply at warmer or cooler temperatures. No development, oviposition, or infection occurred at < 20 C. Fecundity was highest at 25 C, and 50% of the eggs were laid in the first 4-7 days of oviposition. PMID- 19287631 TI - Efficient Preservation of Root-knot and Cyst Nematodes in Liquid Nitrogen. PMID- 19287630 TI - Use of Chitin for Controlling Heterodera avenae and Tylenchulus semipenetrans. AB - The nematicidal effect of chitin, relative to other pesticides, was evaluated against two plant-parasitic nematodes, Heterodera avenae and Tylenchulus semipenetrans. Wheat seedlings, grown in soils artificially or naturally infested with H. avenae, were treated with 0.4% (w/w) ClandoSan (CLA) prepared from crustacean chitin, aldicarb (Temik 15G), or ethylene dibromide (EDB 90EC). The CLA treatment significantly increased wheat straw, ear, and average grain dry weights of nematode-infected plants, compared with the other two treatments. In an experiment covering two consecutive seasons, all three treatments reduced the number of cysts in the soil by 60%. In a one-season experiment, CLA reduced the number of cysts by 51% and aldicarb or EDB reduced cyst number by about 40%. A reduction of 50-90% in T. semipenetrans population densities on roots of two citrus rootstocks was recorded following an application of 0.2% (w/w) CLA to the soil. PMID- 19287632 TI - Neoplastic Growths in Preparasitic Juveniles of Meloidogyne incognita. PMID- 19287633 TI - Parasitism of the Citrus Nematode, Tylenchulus semipenetrans, by Pasteuria penetrans in Iraq. PMID- 19287634 TI - Infectivity of Steinernema feltiae in Fenamiphos-treated Sand. PMID- 19287635 TI - Applications of nematode community structure research to agricultural production and habitat disturbance. AB - Nematode communities in agricultural habitats are diverse, usually comprising tens of species and a large number of individuals. The study of nematode community structure can be approached in a number of ways which can be categorized under a synecological approach. Although the plant-parasitic species are of most obvious interest to plant hematologists, these are only a part of the nematode fauna that affects agroecosystems. The application of community structure research to investigation of those effects can help elucidate their importance. This information is not only intrinsically valuable, but it is necessary in the development of sustainable agricultural production systems. PMID- 19287636 TI - Suggested format for taxonomic papers published in the journal of nematology. PMID- 19287637 TI - Pathological Reaction of Crested Wheatgrass Cultivars to Four Meloidogyne chitwoodi Populations. AB - Meloidogyne chitwoodi populations from Tulelake, California; Ft. Hall, Idaho; Beryl, Utah; and Prosser, Washington, significantly (P < 0.05) reduced dry shoot weights of crested wheatgrass (Agropyron cristatum L., Gaertn. and A. desertorum, Fisch. ex Link, Schult.) cultivars Hycrest, Fairway, and Nordan in experiments conducted in a greenhouse and growth chamber. Shoot growth depression, root galling, and nematode reproduction indices were greatest (P < 0.05) on plants inoculated with 5,000 eggs/plant. Nematode populations from Tulelake, Ft. Hall, and Beryl significantly (P < 0.05) reduced the growth of the three grass cultivars at 15, 20, 25, and 30 C; the greatest reductions occurred at 20 and 25 C. There were significant differences in the virulence of the nematode populations at high (30 C) and low (15 C) soil temperatures. At 15 C, plant growth was reduced more by the Beryl and Tulelake than by the Ft. Hall population; whereas at 30 C, the Ft. Hall population was more virulent than the Beryl and Tulelake populations. Root galling and nematode reproduction were greater on plants inoculated with Beryl and Tulelake populations at 15 C than on plants inoculated with the Ft. Hall population, while the Ft. Hall population had the most pronounced effects at 30 C. PMID- 19287638 TI - Further Details and SEM Observations on Meloidogyne marylandi (Nematoda: Meloidogynidae). AB - Specimens of Meloidogyne marylandi from Bermuda grass and a population from Zoysia grass were examined and compared morphologically by light and electron microscopy. The populations probably are conspecific and the differences noted in the Zoysia population, mainly those of second-stage juveniles (J2) with shorter tails, are considered normal variations rather than representing another form. Scanning electron microscope observations provided additional details of the perineal pattern and head of females and head and lateral fields of second-stage juveniles. Relationship of M. marylandi to closely related species is given. This species is currently known to occur only in Maryland, and populations previously reported from this state as M. graminis are now considered to be M. marylandi. Other reports of M. graminis in the United States now need to be reconfirmed by examination of voucher or recollected specimens. PMID- 19287639 TI - Effects and dynamics of a nematode community on maize. AB - Relationships between nematode density and yield and between final and preplant population levels were examined in small maize plots on sandy soils in north central Florida. Plant-parasitic nematodes present in the community included Belonolaimus longicaudatus, Criconemella sphaerocephala, Meloidogyne incognita, Paratrichodorus minor, Pratylenchus brachyurus, and a Xiphinema sp. Plant growth- including stand count, grain yield, stalk weight, and size of young plants--often was inversely correlated (P 1,000 daltons) excretions secretions (ES) were isolated and partially characterized. The ES consisted of a mixture of proteins, glycoproteins, and proteoglycans or polysaccharides as revealed by differential staining on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and compositional analysis. Carbohydrate, with approximately equal amounts of neutral monosaccharides and hexosamines, was the major constituent of the ES, with only low levels of protein detected. Acidic sugar residues, including sialic acids, were not detected. PMID- 19287642 TI - Carbamate and Organophosphorus Nematicides: Acetylcholinesterase inhibition and Effects on Dispersal. AB - The sensitivities of acetylcholinesterases (ACHE) from the fungus-feeder Aphelenchus avenae and the plant-parasitic species Helicotylenchus dihystera and Pratylenchus penetrans and the housefly, Musca domestica, were compared using a radiometric assay which utilized H(3) acetylcholine as a substrate. Nematode ACHE were generally less sensitive to inhibition by organophosphorns and carbamate pesticides than were ACHE from the housefly. ACHE from the plant-parasitic species and A. avenae were generally similar in sensitivity. In soil, carbamates were more toxic than the organophosphorus pesticides to A. avenae. All pesticides tested affected nematode movement, but fenamiphos was more inhibitory than others. The effects on dispersal of nematodes may be an important mechanism in control by some nematicides. PMID- 19287643 TI - Effects and dynamics of a nematode community on soybean. AB - The relationships between densities of all members of a plant-parasitic nematode community and yield of 'Davis' soybean and between final and preplant population levels were examined in small plots on sandy soils in north-central Florida. Plant-parasitic nematodes present in the community included Belonolaimus longicaudatus, Criconemella sphaerocephala, Meloidogyne incognita, Paratrichodorus minor, Pratylenchus brachyurus, and Xiphinema sp. Plant growth, including stand count, soybean yield (kg/ha), and size of young plants, was occasionally inversely correlated (P 70.0 J2/cm(3) occurred in soil with broomsedge. These findings provide conclusive evidence that T. graminis is a specific parasite of grasses and does not infect citrus. PMID- 19287669 TI - Host Suitability of Soybean Cultivars for Meloidogyne incognita and M. arenaria. AB - The suitability of five maturity group (MG) III and five MG IV soybean, Glycine max, cultivars as hosts for Meloidogyne incognita and M. arenaria was evaluated in a greenhouse. 'Forrest', a MG V cultivar, was used as the standard of comparison for M. incognita resistance. With M. incognita, root-gall and egg-mass indices and reproductive factors for 'Asgrow 3307', 'FFR 398', and 'Pioneer 9442' were comparable with those found on Forrest. Meloidogyne arenaria reproduction was lower (P 75%. When nematicides were removed, many A. avenae exposed to carbofuran resumed normal movement but A. avenae treated with fenamiphos did not recover. Acetylcholinesterase (AChE) activity was suppressed by > 95% in nematodes treated with carbofuran or fenamiphos. However, 48 hours after treated nematodes had been placed in water, AChE activity in carbofuran treated populations was 98% of the levels in control nematodes. Nematodes that had been treated with fenamiphos showed only slight AChE recovery. The antidotes, atropine sulfate and 2-PAM, were largely ineffective in counteracting the toxic effects of the nematicides. PMID- 19287709 TI - Steinernema scapterisci n. sp. (Rhabditida: Steinernematidae). AB - Steinernema scapterisci n. sp., isolated in Uruguay from the mole cricket Scapteriscus vicinus, can be distinguished from other members in the genus by the presence of prominent cheilorhabdions, an elliptically shaped structure associated with the excretory duct, and a double-flapped epitygma in the first generation female. The spicules of the male are pointed, tapering smoothly to a small terminus, and the shaft (calomus) is long, bearing a sheath. The gubernaculum has a long, upward-bent anterior part. The ratio of head to excretory pore divided by tail length of the third-stage juvenile is greater for S. scapterisci n. sp. than for S. carpocapsae. Steinernema scapterisci n. sp. did not hybridize with S. carpocapsae strain Breton. In laboratory tests, S. scapterisci n. sp. killed 10% or less of non-orthopteran insects, including the wax moth larva, a universal host for other species of Steinernema. PMID- 19287710 TI - Trichodorus philipi n. sp. from South Africa, with Notes on Paratrichodorus lobatus and P. acutus. AB - A new species in the family Trichodoridae, from the rhizosphere of native plants of the Cape Fynbos in the western Cape, Republic of South Africa, is described and illustrated. Trichodorus philipi n. sp. resembles T. cottieri Clark, 1963 and T. degans Allen, 1957, from which both sexes are distinguished by a shorter body and onchiostyle. The females differ by the more posterior position of excretory pore, presence of two pairs of lateral body pores, and shape and sclerotization of vulval-vaginal region in lateral view. Males differ by the more posterior position of excretory pore and ventromedian cervical papilla and length and shape of the spicules. Additional morphological details and morphometrics are given for Paratrichodorus lobatus (Colbran, 1965) Siddiqi, 1974 and P. acutus (Bird, 1967) Siddiqi, 1974. PMID- 19287711 TI - Association of Verticillium chlamydosporium and Paecilomyces lilacinus with Root knot Nematode Infested Soil. AB - Population densities of Meloidogyne incognita and the nematophagous fungi, Paecilomyces lilacinus and Verticillium chlamydosporium, were determined in 20 northern California tomato fields over two growing seasons. Paecilomyces lilacinus was isolated from three fields, V. chlamydosporium was isolated from one field, and both fungi were isolated from 12 fields. Verticillium chlamydosporium numbers were positively correlated with numbers of M. incognita and P. lilacinus. Paecilomyces lilacinus numbers were positively correlated with V. chlamydosporium numbers, but they did not correlate with M. incognita numbers. The correlation coefficients were low (R < 0.5) but significant (P < 0.05). All P. lilacinus and V. chlamydosporium field isolates parasitized M. incognita eggs in vitro. In a greenhouse study, numbers of V. chlamydosporium and P. lilacinus increased more in soils with M. incognita-infected tomato plants than in soil with uninfected tomato plants. After 10 weeks, the Pf/ Pi of second-stage juveniles in soils infested with P. lilacinus, V. chlamydosporium, and M. incognita was 47.1 to 295.6. The results suggest V. chlamydosporium and P. lilacinus are not effectively suppressing populations of M. incognita in California tomato fields. PMID- 19287712 TI - Description of Pratylenchoides camachoi n. sp. (Tylenchida: Pratylenchidae) from Spain. AB - Pratylenchoides camachoi n. sp. from rosemary (Rosmarinus officinalis L.) in Sierra De Cazorla in southeastern Spain is described and illustrated. Pratylenchoides camachoi n. sp. is a bisexual species characterized by a cylindroid, sometimes clavate-shaped, male tail with a uniquely shaped smooth, rounded terminus. Females are characterized by a short esophageal overlap and six lines in the lateral field, which reduce to four in the tail region; not areolated except outer bands in the esophageal and tail regions. PMID- 19287713 TI - Coating Soybean Seed with Oxamyl for Control of Heterodera glycines. AB - Oxamyl coated on soybean (Glycine max (L.) Merr. cv. Elgin) seeds in solutions of 20, 40, 80, and 160 mg/ml had no serious deleterious effects on seedling emergence and growth when planted in sterile soil. Seedling emergence on day 3 was less than that of the uncoated control, but by day 7 emergence was equal to, or greater than, the control. Shoot and root growth from seed coated with oxamyl in 40 and 80 mg/ml solutions was greater than that of the control. In soil infested with soybean cyst nematode, Heterodera glycines, shoot weight of soybean plants from seeds coated with oxamyl in 80 mg/ml solution was 11 and 9% greater at weeks 3 and 7, respectively, than from uncoated seeds. Numbers of juveniles (J3 and J4) and adults of H. glycines observed on the roots of plants from oxamyl coated seeds were 83, 42, and 49% less at weeks 3, 5, and 7, respectively, than numbers on the roots of the untreated control. Numbers of J2 extracted from the roots of plants from oxamyl-coated seeds were 75% less at weeks 5 and 7 than those extracted from roots of uncoated seeds. The numbers of J2 extracted from the soil planted to oxamyl-coated seeds were 51 and 33% less at weeks 5 and 7, respectively, than from soil planted to uncoated seed. PMID- 19287714 TI - Physiological Response of Resistant and Susceptible Vitis vinifiera Cultivars to Meloidogyne incognita. AB - The effect of Meloidogyne incognita on growth, general physiological response, and the concentration of reducing and nonreducing sugars at the nematode feeding sites of French Colombard (susceptible) and Thompson Seedless (moderately resistant) Vitis vinifiera cultivars was studied up to 2,100 degree-days (DD-base 10 C). Nematode stress dosage, measured as the product of cumulative number of juveniles and females and their total energy (calories) demand, accounted for up to 15 and 10% of the energy assimilated by French Colombard and Thompson Seedless plants, respectively. Total leaf area, total carbon dioxide fixed, transpiration rate, stomatal conductance, and internal leaf CO concentration were not affected, but energy assimilated into plant tissue and respiration were decreased by nematode infection in both cultivars. Energy consumed by nematodes accounted for most of the difference in total energy assimilated between infected and uninfected plants on French Colombard but not on Thompson Seedless, suggesting that the resistant cultivar may be using more energy to curtail the nematode's activity. Nematodes did not affect the concentration of reducing sugars, but the concentration of nonreducing sugars increased in French Colombard and decreased in Thompson Seedless. This indicates that there was more translocation of photosynthate to the feeding sites of the susceptible than to those of the resistant cultivar, and may explain why M. incognita causes more damage to French Colombard than to Thompson Seedless. PMID- 19287715 TI - Transmission of Pinewood Nematode Through Feeding Wounds of Monochamus carolinensis (Coleoptera: Cerambycidae). AB - Transmission of pinewood nematode, Bursaphelenchus xylophilus, to mature, field grown Scots pines through feeding wounds of Monochamus carolinensis was investigated by caging nematode-infested beetles on pine branches for 24 hours. Nematodes were transmitted to 31 of 64 branches. Frequency of successful transmission was independent of the sex of the beetle but dependent upon beetle age. Transmission frequencies were highest for beetles 2, 3, and 4 weeks after emergence as adults. The number of nematodes transmitted per branch was low and did not differ between beetle sexes or among beetle age categories. The number of nematodes extracted per branch was correlated with the number of nematodes carried per beetle but was not correlated with the feeding area on the branch. PMID- 19287716 TI - Response of Resistant Soybean Plant Introductions to Meloidogyne incognita in Field Microplots. AB - The response of two soybean plant introductions, PI 96354 and PI 417444, highly resistant to Meloidogyne incognita, to increasing initial soil population densities (Pi) (0, 31, 125, and 500 eggs/100 cm(3) soil) of M. incognita was studied in field microplots for 2 years. The plant introductions were compared to the cultivars Forrest, moderately resistant, and Bossier, susceptible to M. incognita. Averaged across years, the yield suppressions of Bossier, Forrest, PI 417444, and PI 96354 were 97, 12, 18, and < 1%, respectively, at the highest Pi when compared with uninfested control plots. Penetration of roots by second-stage juveniles (J2) increased linearly with increasing Pi at 14 days after planting. At the highest Pi, 62% fewer J2 were present in roots of PI 96354 than in roots of the other resistant genotypes. Soil population densities of M. incognita were lower on both plant introductions than on Forrest. At 75 and 140 days after planting, PI 96354 had the lowest number of J2 in the soil, with 49% and 56% fewer than Forrest at the highest Pi. The resistance genes in PI 96354 should be useful in a breeding program to improve the level of resistance to M. incognita in soybean cultivars. PMID- 19287717 TI - Expression of Resistance to Meloidogyne arenaria in Arachis batizocoi and A. cardenasii. PMID- 19287718 TI - Structure of the Cuticle of Metadasynemoides cristatus (Chromadorida: Ceramonematidae). AB - Structure of the cuticle of Metadasynemoides cristatus (Chromadorida: Ceramonematidae) is examined by light, scanning, and transmission electron microscopy. The nematode has more than 600 annuli, and each annulus has eight cuticular plates. Eight longitudinal ridges, beginning on the cephalic capsule, extend the whole length of the body. Where a ridge traverses an annulus, it forms a complicated articulating structure of overlapping vanes. Within the electron dense cortical layer, from which the cuticular plates are formed, there are spaces crossed by fine fibrillae, forming what have been termed "vacuoles" by light microscopists. There is an epicuticle and a continuous lucent basal layer. There appears to be no median layer. The cuticle lining of the esophagus and that forming the circum-oral ridge is of much simpler construction. PMID- 19287719 TI - Xiphinema llanosum and Trophurus vultus, Two New Plant Nematode Species from Pasture Soils in Colombia. AB - Xiphinema llanosum n. sp. and Trophurus vultus n. sp. are described and illustrated from grass soils in Llanos Orientales, Colombia. Xiphinema llanosum is a bisexual species. The female body length is 2.3-2.7 mm, odontostyle 86-96 mum, and odontophore 58-65 mum long; vulva at 42-47%; anterior ovary is absent; the anterior uterus and oviduct are similar to the posterior branch but slightly reduced; and the tail is dorsally convex-conoid with a blunt hemispherical terminus. Male body length is 2.06-2.96 mm; spicules are 40-44 mum long; and four (rarely three or five) anterior ventromedian supplementary papillae are present. Trophurus vultus females are 0.52-0.67 mm long; vulva at 56-60%; stylet is 10.5 13.5 mum long; isthmus is as long as the basal esophageal bulb; the tail is subclavate, 1.6-2.2 times anal body width long; and the terminal cuticle thickness is about one-sixth of the tail length. PMID- 19287720 TI - Histopathology of Ditylenchus destructor on Peanut. AB - The time and mode of entry, and development of Ditylenchus destructor in peanut were studied in field and greenhouse experiments. Few nematodes were present in the cortex of the roots. At 90-120 days after planting, D. destructor was observed in the exocarp at the base of the pod near the point of connection with the peg. The peg was invaded from this primary infection site. The endocarp of the hull was usually penetrated through openings at the base of the mesocarp and sometimes at the pod apex. Numerous D. destructor were present in the testa and the vascular bundles. Nematodes were found in the embryo but not in the cotyledons. The histopathology of D. destructor closely resembles that of the peanut testa nematode, Aphelenchoides arachidis Bos. PMID- 19287721 TI - Optimization of Mitochondrial DNA-based Hybridization Assays to Diagnostics in Soil. AB - Nucleic acid hybridization among root-knot nematode mitochondrial DNAs can be used to identify several Meloidogyne species. Research was initiated to optimize mitochondrial DNA-based molecular diagnostics for the demanding environments likely to be encountered in field isolates. DNA hybridization using reconstituted DNA-soil mixtures revealed a loss of assay sensitivity ranging from 34% to 92% with four agronomic soils tested. This problem was alleviated by the addition of exogenously added DNA. Variation in nematode egg lysis procedures also affected hybridization efficiency, with NaOC1 treatment most effective at disrupting Meloidogyne eggs. These optimized conditions permit detection of mtDNA released from one to five Meloidogyne eggs using standard nucleic acid hybridization procedures. PMID- 19287722 TI - Meloidogyne morocciensis n. sp. (Meloidogyninae), a Root-knot Nematode from Morocco. AB - Meloidogyne morocciensis n. sp. is described from specimens parasitic on peach rootstock from Morocco. This species exhibits a combination of morphological characters similar to M. arenaria, M. incognita, and M. javanica. The perineal pattern of females is oval to squarish with a moderately high to high dorsal arch, and widely spaced, smooth striae; lateral lines are absent. The stylet, 16.5 mum long, has transversely ovoid, set-off knobs. Males have a set-off, annulated head region. The large, rounded labial disc is distinctly demarcated from the crescent-shaped medial lips; lateral lips are absent. The robust stylet, 24.6 mum long, has large, rounded knobs that taper slightly posteriorly. Mean second-stage juvenile (J2) length is 401 mum. The set-offhead region has incomplete annulations; the lip structures are dumbbell shaped. The stylet, 12.3 mum long, has rounded knobs that slope posteriorly. The J2 tail, 52.6 mum long, has irregularly sized annules in the posterior region and ends in a bluntly rounded tip. Tomato, tobacco, pepper, and watermelon are good hosts; cotton and peanut are not hosts. Meloidogyne morocciensis n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of 47-49. Its esterase phenotype is identical with the three-banded phenotype (A3) of M. arenaria. PMID- 19287723 TI - Host Status of Seven Weed Species and Their Effects on Ditylenchus destructor Infestation of Peanut. AB - The host suitability to Ditylenchus destructor of seven common weed species in peanut (Arachis hypogaea) fields in South Africa was determined. Based on the number of nematodes per root unit, white goosefoot (Chenopodium album), feathertop chloris (Chloris virgata), purple nutsedge (Cyperus rotundus), jimson weed (Datura stramonium), goose grass (Eleusine indica), khaki weed (Tagetes minuta), and cocklebur (Xanthium strumarium) were poor hosts. Ditylenchus destructor survived on all weed species; population densities increased in peanut hulls and caused severe damage to seeds of peanut grown after weeds. Roots of purple nutsedge left in the soil suppressed populations of D. destructor and root and pod development in peanut grown after the weed. However, nematode populations in peanut hulls and seeds were not suppressed. Some weed species, especially purple nutsedge which is common in peanut fields, can be used to indicate the presence of D. destructor in the absence of peanut. PMID- 19287724 TI - Relationship of Bursaphelenchus xylophilus Population Density to Mortality of Pinus sylvestris. AB - Seven-month-old Scots pine seedlings were inoculated with water or culture filtrate (controls), with 10,000, or 20,000 (experiment 1), and with 2,500 (experiment 2) Bursaphelenchus xylophilus B.C. isolate nematodes and maintained under defined experimental conditions. Controls did not develop pine wilt disease over a 2-month period. In experiment 1, less than 50% of the inoculum was recovered from the nematode-inoculated seedlings in the first 48 hours, after which the nematode population of both treatments increased exponentially resulting in pine death and approximately equal populations at 216 hours after inoculation. In the second experiment, plant mortality, which was always preceded by 2-3 days of chlorosis and associated stem vascular necrosis, first occurred 14 days after inoculation. The nematode population increased until about day 40 after inoculation and declined thereafter. Nematodes extracted from the roots 2 weeks after inoculation accounted for ca.15% of the total number of nematodes per pine. The study indicates that the rate of nematode reproduction is a factor in pine wilt disease. However, the lack of a linear correlation between the number of nematodes and the timing of pine mortality suggests that the timing of pine death may also depend on the location of nematode damage to the host tissue. PMID- 19287725 TI - Reproduction and Parasitism of Pratylenchus neglectus on Potato. AB - An initial density (Pi) of 1,540 Pratylenchus neglectus/kg soil suppressed shoot growth of potato, Solanum tuberosum cv. Russet Burbank, in a greenhouse test at 3 weeks. After 6 weeks, shoot weights were reduced by Pi of 662 and 1,540 nematodes/kg soil, the final soil densities of P. neglectus were twice the respective Pi, and the numbers of nematodes per gram dry root were 5,363 and 7,981. In 1986-88 field microplot experiments with the Norchip cultivar, neither shoot nor root weight was suppressed by P. neglectus. In 1986 a Pi of 115 nematodes/kg soil suppressed the total number and weight of tubers per plant. In 1987 a Pi of 186 nematodes/kg soil suppressed the marketable and total number of tubers by 19 and 25 %, respectively. In 1988 a Pi of 1,884 nematodes/ kg soil reduced total and marketable weight by 18 and 19%, respectively. In 1986 and 1987 nematode population densities in the soil increased 34-fold and 27-fold, respectively. In 1988 the Pi of 1,884 nematodes/kg soil rose to 21,890/kg at midseason, then dropped to 4,370/kg at harvest. These studies show for the first time that P. neglectus reproduces well on potato and can cause yield losses. Because of its distribution and abundance, P. neglectus may be considered an economically important parasite of potato in Ontario. PMID- 19287726 TI - Observations on the Invasion and Endoparasitic Behavior of the Root Lesion Nematode Pratylenchus penetrans. AB - The endoparasitic behavior of Pratylenchus penetrans was examined using video enhanced contrast microscopy to observe living nematodes inside root tissue. Feeding behavior could be separated into phases of probing, cell penetration by the stytet, salivation, and food ingestion for brief and extended periods. After cell penetration, a small "salivation zone" was formed around the stylet tip. No feeding tubes were observed. Feeding and migration were interrupted by rest phases when a nematode became characteristically coiled inside a cell. Tissue damage was caused primarily by migration and extended feeding periods. Aspects of egg laying and molting are also described. PMID- 19287727 TI - In Vitro Embryo Explant Cultures of Peanut to Evaluate Resistance to Ditylenchus destructor. AB - Population densities of D. destructor on embryo explants of 22 peanut genotypes grown in vitro were compared with those in roots and seeds of the same genotypes grown in the greenhouse. During the first 8 weeks after inoculation, the optimum incubation period was 6 weeks for maximum reproduction of Ditylenchus destructor on embryo explants of peanut (Arachis hypogaea L. cv. Sellie) inoculated with 250 nematodes at 25 C. Nematode numbers increased 17-fold. Deletion of MnSO HO and a higher KHPO concentration in the medium resulted in higher nematode reproduction. Resistance or susceptibility to D. destructor was observed in seeds of several genotypes but was not matched by differences in host suitability in roots. The results indicate that the factor for resistance or susceptibility to D. destructor is synthesized in the seeds of peanut but is not translocated to the roots. Use of embryo explant cultures of peanut as a rapid method to evaluate resistance to D. destructor did not work under the conditions described. PMID- 19287728 TI - Artificial feeding systems for plant-parasitic nematodes. AB - Feeding of an "obligate" plant-parasitic nematode (nonfungal feeder), Pratylenchus scribneri, in the absence of plant tissue was demonstrated in an artificial system consisting of liquid media and indicator dyes including amaranth and various nontoxic food colors. Among the compounds tested, sucrose, dextrose, Gamborg's B5 medium, and DL-methionine stimulated a small percentage of feeding (12-36%). A high percentage of feeding (90-100%) occurred in a filtrate from excised corn roots cultured in Gamborg's B5 medium. This feeding system has the potential to develop an artificial medium for plant-parasitic nematodes and to screen novel nematicides that are stomach poisons. PMID- 19287729 TI - Pathological Relationship of Ditylenchus dipsaci and Fusarium oxysporum f. sp. medicaginis on alfalfa. AB - Ditylenchus dipsaci and Fusarium oxysporum f. sp. medicaginis synergistically affected the mortality and plant growth of Ranger alfalfa, a cultivar susceptible to stem nematode and Fusarium wilt. The nematode-fungus relationship had an additive effect on mortality and plant growth of Lahontan (nematode resistant and Fusarium wilt susceptible) and of Moapa 69 (nematode susceptible and Fusarium wilt resistant). Mortality rates were 13, 16, 46, and 49% for Ranger; 4, 18, 26, and 28% for Lahontan; and 19, 10, 32, and 30% for Moapa 69 inoculated with D. dipsaci, F. oxysporum f. sp. medicaginis, and simultaneously and sequentially with D. dipsaci and F. oxysporum f. sp. medicaginis, respectively. Shoot weights as a percentage of uninoculated controls for the same treatments were 52, 84, 26, and 28%, for Ranger; 74, 86, 64, and 64% for Lahontan; and 50, 95, 44, and 39% for Moapa 69. Plant growth suppression was related to vascular bundle infection and discoloration of alfalfa root tissue. Disease severity and plant growth of alfalfa did not differ with simultaneous or sequential inoculations of the two pathogens. Fusarium oxysporum f. sp. medicaginis affected alfalfa growth but not nematode reproduction. PMID- 19287730 TI - Teratorhabditis palmarum n. sp. (Nemata: Rhabditidae): An Associate of Rhynchophorus palmarum and R. cruentatus. AB - Teratorhabditis palmarum n. sp., an associate of the palm weevils Rhynchophorus palmarum and R. cruentatus is described. Teratorhabditis palmarum was isolated from newly emerged adults and cocoons of R. palmarum from red-ring diseased coconut palms, Cocos nucifera, in Trinidad and Ecuador, and from red-ring diseased oil palms, Elaeis guineensis, in Colombia. Teratorhabditis palmarum was also associated internally with newly emerged adults of R. cruentatus from mature transplanted cabbage palmettos, Sabal palmetto, in Florida. Dauer juveniles of T. palmarum infested the genital capsule and body cavity of newly emerged adult female and male palm weevils. Adult nematodes isolated from cocoons and dauer juveniles from newly emerged palm weevils were cuhurable on bacterial lawns on several solid media. Females of T. palmarum have a single anteriorly directed ovary; vulva at 93-96% of the body length; short, hemispherical spicate tail; three or four teeth in the metastom; cuticle with distinct transverse punctations that change abruptly at the level of the procorpus to indentations of alternating size and arrangement; and eggs with cuticular sculpturing. Males have a crenate, peloderan bursa with a 2 + 5 + 3 pattern of bursal rays (7 extend to the margin of bursa); spicules linear, completely fused at the distal tip and dorsally for 50% of the total spicule length. PMID- 19287731 TI - Effects of Planting Date, Small Grain Crop Destruction, Fallow, and Soil Temperature on the Management of Meloidogyne incognita. AB - The effects of planting date, rye (Secale cereale cv. Wren Abruzzi) and wheat (Triticura aestivum cv. Coker 797), crop destruction, fallow, and soil temperature on managing Meloidogyne incognita race 1 were determined in a 2-year study. More M. incognita juveniles (J2) and egg-producing adults were found in roots of rye planted 1 October than in roots of rye planted 1 November and wheat planted 1 November and 1 December. Numbers of M. incognita adults with and without egg masses were near or below detectable levels in roots of rye planted 1 November and wheat planted 1 November and 1 December. Meloidogyne incognita survived the mild winters in southern Georgia as J2 and eggs. The destruction of rye and wheat as a trap crop 1 March suppressed numbers of J2 in the soil temporarily but did not provide long-term benefits for susceptible crops that followed. In warmer areas where rye and wheat are grown in winter, reproduction of M. incognita may be avoided by delaying planting dates until soil temperature declines below the nematode penetration threshold (18 C), but no long-term benefits should be expected. The temperature threshold may be an important consideration in managing M. incognita population densities in areas having lower winter soil temperatures than southern Georgia. PMID- 19287732 TI - Effect of Age on Body Wall Cuticle Morphology of Heterodera schachtii Schmidt females. AB - Fine structure of the body wall cuticle of Heterodera schachtii is compared with respect to age and body region of the female. The cuticle is more complex than previously reported. In newly molted females only layers A, B, and C are present, but 4 weeks after the final molt a thin D layer is present between the midbody and base of the cone. This D layer is absent in the cone of H. schachtii, regardless of age. As females age, an additional layer E is produced and includes zones E and E. Zone El apparently is unique to H. schachtii, whereas E is likely to be homologous with a similar layer in Atalodera. In the cone of old females (ca. 8 weeks after the final molt) of H. schachtii, the two zones become irregular in shape and comprise bullae. The presence of a thin D layer in Heterodera strengthens the previous hypothesis of a single ancestor of cyst nematodes. PMID- 19287733 TI - Ultrastructure of Phasmid Development in Meloidodera floridensis and M. charis (Heteroderinae). AB - Phylogenetic characters for Heteroderinae Luc. et al., 1988 are evaluated in Meloidodera which is believed to have primarily ancestral characters. Phasmid ultrastructure is observed in second-stage juveniles (J2), third-stage juvenile males, fourth-stage juvenile males, and fifth-stage males of Meloidodera floridensis and M. charis. Phasmid secretion occurs inside the egg before the J1 J2 molt. Before J2 hatch, concentric lamellar membranes occur within the sheath and socket cells. Some membranes become lamellae of the sheath cell plasma membrane; others become multilamellar bodies. During early molting, plasma membrane lamellae disappear and a distal dendrite segment appears in a rudimentary canal. After the molt, the distal dendrite is not present within the canal. The phylogenetic utility of phasmid features is discussed. In both species the ampulla shape and size between molts are stable features in juveniles and males. The posthatch J2 sheath cell receptor cavity may vary in a species specific manner, but comparative morphology requires precise timing after hatch. PMID- 19287734 TI - Phylogenetic implications of phasmid absence in males of three genera in heteroderinae. AB - Absence of the phasmid was demonstrated with the transmission electron microscope in immature third-stage (M3) and fourth-stage (M4) males and mature fifth-stage males (M5) of Heterodera schachtii, M3 and M4 of Verutus volvingentis, and M5 of Cactodera eremica. This absence was supported by the lack of phasmid staining with Coomassie blue and cobalt sulfide. All phasmid structures, except the canal and ampulla, were absent in the postpenetration second-stage juvenile (J2) of H. schachtii. The prepenetration V. volvingentis J2 differs from H. schachtii by having only a canal remnant and no ampulla. This and parsimonious evidence suggest that these two types of phasmids probably evolved in parallel, although ampulla and receptor cavity shape are similar. Absence of the male phasmid throughout development might be associated with an amphimictic mode of reproduction. Phasmid function is discussed, and female pheromone reception ruled out. Variations in ampulla shape are evaluated as phylogenetic character states within the Heteroderinae and putative phylogenetic outgroup Hoplolaimidae. PMID- 19287735 TI - Reproduction of Globodera rostochiensis on Transformed Roots of Solanum tuberosum cv. Desiree. AB - Transformed roots of the susceptible potato Solanum tuberosum L. cv. Desiree were inoculated with second-stage juveniles (J2) of Globodera rostochiensis pathotype Rol. Adult males emerged after 3-4 weeks and matings with females occurred. After 8 weeks gentle pressure on the eggs of maturing females released the J2 which were viable. Because this technique enables the production of vigorously growing roots with numerous laterals, it may be suitable for obtaining a high yield of sterile potato cyst nematodes. PMID- 19287736 TI - Improved nematode extraction from carrot disk culture. AB - Radopholus spp. were reared in carrot tissue culture via established procedures, with slight modification. Several plant tissue maceration enzymes and flotation media (salts and sucrose) were evaluated with regard to nematode toxicity and extraction efficiency. Best extraction of viable nematodes and eggs was attained when carrot tissue infested with Radopholus citrophilus or R. similis was macerated with a mixture of 0.50% driselase and 0.50% cellulysin, w/v each, with 2.5 ml of enzyme solution based for each gram of carrot tissue. Maceration slurries containing carrot tissue and nematodes were maintained in open flasks on a rotary shaker (175 rpm) at 26 C for 24 hours. Nematodes and eggs were extracted from resultant culture slurries by flotation with MgSO-7H0 (sp gr 1.1). A protocol is presented to extract large quantities of viable burrowing nematodes and their eggs from carrot disk cultures. PMID- 19287737 TI - A digital image analysis system for comparing groups of small nematodes. AB - A digital imaging system was developed for measuring various physical characteristics of individual nematodes and for comparing groups of nematodes. The equipment consists of a microscope, a video camera, a video digitizer, interactive displays, and a computer. Various physical and mathematical methods were incorporated, algorithms devised, and computer software written for image acquisition, editing, and analysis. To test the system, four populations of an isolate of the pinewood nematode, Bursaphelenchus xylophilus, subjected to 100% relative humidity at 22 C for 0, 12, 24, or 48 hours were compared. The results showed that the system can be used to measure physical parameters of individual nematodes and to differentiate groups of nematodes. PMID- 19287738 TI - Temperature and the Life Cycle of Heterodera zeae. AB - Development of the corn cyst nematode, Heterodera zeae, was studied in growth chambers at 20, 25, 29, 33, and 36 +/- 1 C on Zea mays cv. Pioneer 3184. The optimum temperature for reproduction appeared to be 33 C, at which the life cycle, from second-stage juvenile (J2) to J2, was completed in 15-18 days; at 36 C, 19-20 days were required. Juveniles emerged from eggs within 28 days at 29 C and after 42 days at 25 C. Although J2 were present within eggs after 63 days at 20 C, emergence was not observed up to 99 days after inoculation. Female nematodes produced fewer eggs at 20 C than at higher temperatures. PMID- 19287739 TI - Effects of Carbendazim on the Nematophagous Fungus Hirsutella rhossiliensis and the Ring Nematode. PMID- 19287740 TI - Evaluation of Entomopathogenic Nematodes against the Mexican Rice Borer (Lepidoptera: Pyralidae). PMID- 19287741 TI - Expression of Resistance to Meloidogyne arenaria in Arachis batizocoi and A. Cardenasii. PMID- 19287742 TI - Stability and characteristics of spatial description parameters for nematode populations. AB - The parameters of Taylor's Power Law (s(2) = am(b)) relating variance (s(2)) to mean population level (m) were acceptably stable in different fields with similar cropping systems. Values of both a and b parameters varied with nematode species. The value of a was a function of sample size (number of cores) and was characterized for each species. The value of b was stable across sample size and reflective of the life history strategy of the species. The relationship between the economic threshold and sampling intensity required to allow management decisions, with specified levels of risk, indicated the need for improved sampling technology. PMID- 19287743 TI - Influence of Soil pH and Oxygen on Persistence of Steinernema spp. AB - Survival of infective juveniles of Steinernema carpocapsae and Steinernema glaseri gradually declined during 16 weeks of observation as the tested soil pH decreased from pH 8 to pH 4. Survival of both species of Steinernema dropped sharply after 1 week at pH 10. Survival or S. carpocapsae and S. glaseri was similar at pH 4, 6, and 8 during the first 4 weeks, but S. carpocapsae survival was significantly greater than S. glaseri at pH 10 through 16 weeks. Steinernema carpocapsae and S. glaseri that had been stored at pH 4, 6, and 8 for 16 weeks, and at pH 10 for 1 or more weeks were not infective to Galleria mellonella larvae. Steinernema carpocapsae survival was significantly greater than that of S. glaseri at oxygen:nitrogen ratios of 1:99, 5:95, and 10:90 during the first 2 weeks, and survival of both nematode species declined sharply to less than 20% after 4 weeks. Survival of both nematode species significantly decreased after 8 weeks as the tested oxygen concentrations decreased from 20 to 1%, and no nematode survival was recorded after 16 weeks. Steinernema carpocapsae pathogenicity was significantly greater than that of S. glaseri during the first 2 weeks. No nematode pathogenicity was recorded at oxygen concentrations of 1, 5, and 10% after 2 weeks and at 20% after 16 weeks. PMID- 19287744 TI - Effect of Temperature on Expression of Resistance to Meloidogyne spp. In Common Bean (Phaseolus vulgaris). AB - The effect of soil temperature on the expression of resistance in several common bean lines carrying resistance to root-knot nematodes (Meloidogyne spp.) was studied under controlled temperatures in temperature tank and growth chamber conditions. Resistance to M. javanica and M. incognita race 1 in bean lines A315, A328, A445, G1805, and G2618 was stable at 24-30 C. However, there was a significant increase in reproduction of M. javanica on A315, A328, and A445 when temperature was increased from 26 to 30 C. This increase did not reflect a change from a resistant to a susceptible reaction or classification. Resistance in A315 is derived from G1805, whereas resistance in A328 and A445 is derived from G2618. Alabama No. 1, PI 165426, and PI 165435, with resistance to M. incognita race 2, were heat stressed at temperatures above 27 C. Resistance to M. incognita race 2 in Alabama No. 1 and PI 165435 was lost at 30 C, but PI 165426 supported low reproduction of M. incognita race 2 at all temperatures. Poor root development at 30 C may have been responsible, in part, for the poor development of M. incognita race 2 on PI 165426. PMID- 19287745 TI - Demonstration of Multiple Mating in Heterodera glycines with Biochemical Markers. AB - Controlled crosses of Heterodera glycines were carried out by placing one o r more virgin females of known esterase phenotype on an agar plate and adding, at various time intervals, one or more males of different esterase phenotypes. Progeny (second-stage juveniles) of such crosses were propagated on soybeans, and 30 days later young females were subjected to electrophoretic analysis to determine their esterase phenotype. Esterase phenotypes that represented the heterozygous state of the maternal and paternal genomes confirmed the hybrid nature of the progeny and identified their male parent. When each of 74 females was given the opportunity to mate successively with two males of different esterase phenotypes, 43 mated with a single male and 31 mated with both males. One female mated with three males, i.e., with a male of its own population (sib mating) and the two males provided for the cross. Inseminated females could mate for a second time soon after, or as late as 24 hours after, their first mating. When single males were given the opportunity to mate with many females, about equal numbers of them inseminated zero, one, two, or three females. In greenhouse tests, 12 females were given the opportunity to mate with many males of three different esterase phenotypes. Two females mated with one and possibly more males of the same phenotype, and 10 females mated with males of two different esterase phenotypes. In conclusion, multiple mating appears to be a common behavior of males and females of H. glycines. PMID- 19287746 TI - Descriptions and Comparative Morphology of Cactodera milleri n. sp. (Nematoda: Heteroderidae) and Cactodera cacti with a Review and Key to the Genus Cactodera. AB - A new species, Cactodera milleri n. sp., is described and illustrated from specimens obtained from roots of common lambsquarter, Chenopodium album L., from Mattawan, Michigan. Cactodera milleri can be differentiated from other Cactodera species by the presence of punctated egg shells and a second-stage juvenile (J2) stylet length averaging 21.8 mum as measured from freshly killed specimens in water mounts. Thirty-four plant species, including 11 weed species, 18 agronomic crop species, and 5 cactus species were tested as potential hosts of C. milleri. The new species reproduced only on Chenopodium album, C. amaranticolor Cofte. Reyn., and C. quinoa Willd. Cactodera cacti (Filipjev &Schuurmans Stekhoven, 1941) Krall &Krall, 1978, a morphologically similar species, is reexamined. A description of the female and additional morphometric and morphologic data of cysts, males, J2, and eggs are provided for several populations of C. cacti. A review of the morphometrics of all species of Cactodera and a taxonomic key to the seven species are presented. PMID- 19287747 TI - Plant-parasitic Nematode Acetylcholinesterase Inhibition by Carbamate and Organophosphate Nematicides. AB - The sensitivity of acetylcholinesterases (ACHE) isolated from the plant-parasitic nematodes Meloidogyne arenaria, M. incognita, and Heterodera glycines and the free-living nematode Caenorhabditis elegans to carbamate and organophosphate nematicides was examined. The AChE from plant-parasitic nematode species were more sensitive to carbamate inhibitors than was AChE from C. elegans, but response to the organophosphates was approximately equivalent. The sulfur containing phosphate nematicides were poor inhibitors of nematode acetylcholinesterase, but treatment with an oxidizing agent greatly improved inhibition. Behavioral bioassays with living nematodes revealed a poor relationship between enzyme inhibition and expression of symptoms in live nematodes. PMID- 19287748 TI - Tests for Transmission of Prunus Necrotic Ringspot and Two Nepoviruses by Criconemella xenoplax. AB - In two of three trials, detectable color reactions in ELISA for Prunus necrotic ringspot virus (PNRSV) were observed for Criconemella xenoplax handpicked from the root zone of infected peach trees. Criconemella xenoplax (500/pot) handpicked from root zones of peach trees infected with PNRSV failed to transmit the virus to cucumber or peach seedlings. The nematode also failed to transmit tomato ringspot (TomRSV) or tobacco ringspot viruses between cucumbers, although Xiphinema americanum transmitted TomRSV under the same conditions. Plants of peach, cucumber, Chenopodium quinoa, and Catharanthus roseus were not infected by PNRSV when grown in soil containing C. xenoplax collected from root zones of PNRSV-infected trees. Shirofugen cherry scions budded on Mazzard cherry seedling rootstocks remained symptomless when transplanted into root zones of PNRSV infected trees. Virus transmission was not detected by ELISA when C. xenoplax individuals were observed to feed on cucumber root explants that were infected with PNRSV and subsequently fed on roots of Prunus besseyi in agar cultures. Even if virus transmission by C. xenoplax occurs via contamination rather than by a specific mechanism, it must be rare. PMID- 19287749 TI - Interactions Between Calonectria crotalariae and Heterodera glycines on Soybean. AB - The interactions of Heterodera glycines at four egg inoculum levels (0, 100, 1,000, and 10,000 per pot) and three cyst levels (0, 100, and 200 per pot) and Calonectria crotalariae at 500, 5,000, and 50,000 microsclerotia per pot were evaluated on soybean. At the two lowest nematode egg levels, the presence of C. crotalariae did not affect nematode reproduction. At 10,000 eggs per pot, however, nematode reproduction was increased significantly at each microsclerotial level. The increase in nematode reproduction was stepwise at 500 and 5,000 microsclerotia per pot but declined at 50,000 microsclerotia per pot. Similar results were obtained when cysts rather than eggs were used as nematode inoculum. The nematode x fungus interaction significantly affected 60-day plant growth parameters of both Lee 74 and Centennial soybean. The nematode x fungus interaction was antagonistic to plant roots and significantly influenced root injury ratings. The presence of C. crotalariae in tissues of stock plants or plants used as race differentials did not alter the analysis of this population as race 3. PMID- 19287750 TI - Effect of Bursaphelenchus xylophilus on the Assimilation and Translocation of 14 C in Pinus sylvestris. AB - The effect of wound, wound + water, wound + Bursaphelenchus xylophilus culture filtrate, or wound + lethal B. xylophilus doses on the assimilation and translocation of (1)C by 8-month-old Pinus sylvestris seedlings was tested. In two separate experiments, pine seedlings were exposed to 28.35 muCi of (1)CO for 20 minutes below or above (to the pine shoot leader) the point of nematode inoculation. After 2 and 4 hours of dark adaptation, 80% ethanol soluble (1)C tissue extracts were determined by liquid scintillation counting. Nematode infection significantly (P = 0.05) decreased (1)C assimilation. Treatments translocated less than 6% of the total amount of the fixed (1)C and translocation generally decreased with increasing size of nematode inoculum. However, infected pines translocated a greater proportion of the amount of (1)C fixed per gram of exposed nematode-plant tissue than did the control pines. The lower levels of photoassimilate entering the plant system probably resulted in a reduced metabolic capacity in B. xylophilus-infected pine seedlings. The effect on photosynthesis could be one of the key factors leading to death of pines through starvation, and it is possible that it was preceded by an effect on related physiological processes such as water uptake. PMID- 19287751 TI - Parasitism of Brown Planthopper and Whitebacked Planthopper by Agamermis unka in Korea. AB - Adults of the brown planthopper (BPH) and the whitebacked planthopper (WBPH) that migrated into Korea from China were not parasitized by the mermithid, Agamermis unka. BPH and WBPH collected from Korean rice fields were parasitized. Parasitism of BPH in the lst-3rd, 4th, and 5th instars, and adults was 31.5%, 61.5%, 66.4%, and 45.5%, respectively, whereas parasitism of the same stages of WBPH was 50%, 50%, 100% and 90.7%, respectively. Parasitism of BPH by A. unka significantly reduced the number of eggs. Only 4.2% of the parasitized females contained eggs, whereas 85.6% of unparasitized females had eggs. Tilling of rice fields significantly increased mermithid parasitism of BPH. Mermithids parasitized 39.3% of caged adults in the untilled field and 77.8% in the tilled field. PMID- 19287752 TI - Identification of single meloidogyne juveniles by polymerase chain reaction amplification of mitochondrial DNA. AB - Polymerase chain reaction (PCR) was used to amplify a specific 1.8-kb sequence of mitochondrial DNA from single juveniles and eggs from 17 populations of Meloidogyne incognita, M. hapla, M. javanica, and M. arenaria. Approximately 2 mug amplified product were produced per reaction. Restriction digestion of the amplified product with HinfI permitted discrimination of clonal lineages of the four species. Meloidogyne javanica, however, could not be separated from M. hapla by the enzymes used in these experiments. Various amplification conditions and nematode lysis procedures were examined in order to optimize the speed and quality of identifications. PMID- 19287753 TI - Host Range and Ecology of Isolates of Pasteuria spp. from the Southeastern United States. AB - Isolates of Pasteuria penetrans were evaluated for ecological characteristics that are important in determining their potential as biological control agents. Isolate P-20 survived without loss of its ability to attach to its host nematode in dry, moist, and wet soil and in soil wetted and dried repeatedly for 6 weeks. Some spores moved 6.4 cm (the maximum distance tested) downward in soil within 3 days with percolating water. The isolates varied greatly in their attachment to different nematode species and genera. Of five isolates tested in spore-infested soil, three (P-104, P-122, B-3) attached to two or more nematode species, whereas B-8 attached only to Meloidogyne hapla and B-I did not attach to any of the nematodes tested. In water suspensions, spores of isolate P-20 attached readily to M. arenaria but only a few spores attached to other Meloidogyne spp. Isolate P 104 attached to all Meloidogyne spp. tested but not to Pratylenchus scribneri. Isolate B-4 attached to all species of Meloidogyne and Pratylenchus tested, but the rate of attachment was relatively low. Isolate P-Z00 attached in high numbers to M. arenaria when spores were extracted from females of this nematode; when extracted from M. javanica females, fewer spores attached to M. arenaria than to M. javanica or M. incognita. PMID- 19287754 TI - Isolation of Fungi from Heterodera glycines and in vitro Bioassays for Their Antagonism to Eggs. AB - Twenty fungi were assayed in vitro for antagonism to eggs of Heterodera glycines. Eight of the fungi were isolated from cysts or eggs of H. glycines during the current study, one was isolated from Panagrellus redivivus, and eleven were obtained from other researchers or collections. The bioassays were conducted on eggs from nematodes that had been grown monoxenically on excised root tips. Phoma chrysanthemicola, one strain of Verticillium chlamydosporium, and one strain of V. lecanii caused a decrease (P < 0.01, P < 0.05, P < 0.05, respectively) in the number of viable eggs, although no hyphae were observed colonizing live eggs. Trichoderma polysporum infected live eggs but enhanced (P < 0.05) egg survival. Acremonium bacillisporum, Chaetomium sp., Drechmeria coniospora (two strains), Epicoccum sp., Exophiala jeanselmei, Fusarium sp., Neocosmospora vasinfecta, Scytalidium fulvum, Trichoderma harzianum (two strains), V. chlamydosporium (one strain), V. lecanii (three strains), and an unidentified fungus did not measurably affect egg viability, even though hyphae of five of these fungi were seen in live eggs. The bioassay provides a useful step in the selection of a biological control agent for this major nematode pest. PMID- 19287755 TI - Comparison of Populations of Pratylenchus brachyurus Based on Isozyme Phenotypes. AB - Enzymes from females of five Pratylenchus brachyurus populations and one P. scribneri population were analyzed by isoelectric focusing electrophoresis. Of the 18 enzyme systems investigated, only malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were detected from all five P. brachyurus populations and P. scribneri. Faint bands were detected for isocitrate dehydrogenase and phosphogluconate dehydrogenase from one P. brachyurus population. Three distinct phenotypic groups were found in the MDH and PGM systems for P. brachyurus populations, but only a single electromorph was detected for PGI. Multiple electromorphs for MDH, PGM, and PGI were detected for P. scribneri; there was no similarity among these patterns with those from P. brachyurus. No phenotypic differences in PGI were observed between females and mixed juveniles of one population of P. brachyurus. PMID- 19287756 TI - Biology and Pathogenicity of Pratylenchus neglectus on Alfalfa. AB - Pratylenchus neglectus reduced the growth of alfalfa cultivars in greenhouse and growth chamber studies. Inocula (1,000, 5,000 and 10,000 nematodes per plant) reduced shoot dry weights of Ranger by 16, 27, and 40%, of Lahontan by 16, 32, and 40%, and of Nevada Synthetic XX (Nev Syn XX) by 18, 26, and 37%, respectively, at 26 n 2 C. Pratylenchus neglectus at 1,000 nematodes per plant reduced Ranger shoot dry weights by 5, 12, 18, and 27%, at 15, 20, 25, and 30 C, respectively, whereas 5,000 nematodes per plant reduced shoot dry weights by 12, 17, 26, and 38%, respectively, at similar temperatures. Reductions in dry root weights were directly related to reductions in shoot growth. At 1,000 nematodes per plant, Ranger root dry weights were reduced by 3, 14, 40, and 40%, whereas 5,000 nematodes per plant reduced root dry weight by 25, 31, 59, and 63%, respectively, at similar temperatures. Similar results were observed on Lahontan and Nev Syn XX at the same inoculum levels and soil temperatures. Nematode reproductive indices (final nematode population per plant divided by initial nematode inoculum per plant) were higher at 1,000 nematodes per plant than at 5,000 nematodes per plant, were positively correlated with temperature, and were unaffected by cultivar. PMID- 19287757 TI - Repeated sampling to determine the precision of estimating nematode population densities. AB - The first phase of this study involved repeated samplings of five fields using composite samples of 10, 20, 40, and 80 soil cores, to determine the precision of nematode assays. The second phase focused on randomly selecting two and four 2-ha subunits (data on Meloidogyne spp.) of 24 fields ranging from 6 to 40 ha and computing the precision of estimated means for these numbers ofsubunits versus the general field mean (based on all 2-ha subunits). Average numbers of nematodes from most samples containing Meloidogyne spp., Heterodera glycines, Helicotylenchus dihystera, Scutellonema brachyurum, and (or) Hoplolaimus galeatus were within 50% of the overall means. Coefficient of variation (CV) values were generally lower for 40 cores than for 10, 20, and 80 cores per sample. When data for all nematodes and fields were combined, this value was lowest for 40 and 80 cores. The CV values were higher for Meloidogyne spp. than for H. glycines. Means of two samplings increased the probability of obtaining numbers nearer the mean for that field than numbers from a single composite sample. For the second phase, population estimates of Meloidogyne spp. based on four 2-ha subunits generally were closer to field means than were those for two subunits. Sampling precision with these subunits diminished greatly in large fields with variable soils and (or) mixed cropping histories. Either two or four subunits gave population estimates within 3-20% of the field mean in most instances. The mean man hours required for sampling ca. 2-ha parcels of 4-20-ha fields was 0.54 hours. PMID- 19287758 TI - Morphometrics and SEM Illustrations of Three Species of Ogma Southern, 1914 (Nematoda: Criconematidae) from Spain. AB - Ogma civellae (Steiner, 1949) Raski &Luc, 1987, O. cobbi (Micoletzky, 1922) Siddiqi, 1986, and O. palmatum (Siddiqi &Southey, 1962) Siddiqi, 1986 are reported in natural habitats in southeastern Spain. Each is briefly described using light microscopy, and the original descriptions are amplified and supplemented by scanning electron microscopy (SEM) observations. Measurements with standard deviations and SEM illustrations are included for each species and compared with previous data. PMID- 19287759 TI - Antagonists of Plant-parasitic Nematodes in Florida Citrus. AB - In a survey of antagonists of nematodes in 27 citrus groves, each with a history of Tylenchulus semipenetrans infestation, and 17 noncitrus habitats in Florida, approximately 24 species of microbial antagonists capable of attacking vermiform stages of Radopholus citrophilus were recovered. Eleven of these microbes and a species of Pasteuria also were observed attacking vermiform stages of T. semipenetrans. Verticillium chlamydosporium, Paecilomyces lilacinus, P. marquandii, Streptomyces sp., Arthrobotrys oligospora, and Dactylella ellipsospora were found infecting T. semipenetrans egg masses. Two species of nematophagous amoebae, five species of predatory nematodes, and 29 species of nematophagous arthropods also were detected. Nematode-trapping fungi and nematophagous arthropods were common inhabitants of citrus groves with a history of citrus nematode infestation; however, obligate parasites of nematodes were rare. PMID- 19287760 TI - Vertical Dispersal of Steinernema scapterisci. AB - When infective juveniles ofSteinernema scapterisci Nguyen &Smart were released on the soil surface in the field and in the laboratory, some of them moved downward through the soil at least 10 cm in 5 days and infected and killed mole crickets. When released 2 cm below the soil surface, most of the juveniles moved into the upper 2 cm layer of soil, but some moved downward 10 cm. When placed at the center of a 16-cm soil column, infective juveniles moved in both directions with three times more moving downward than upward. Infective juveniles were more efficient in killing mole crickets in the field than in the laboratory. PMID- 19287761 TI - Paralongidorus bullatus n. sp. from Groundnut Soils in Niger and Comments on Xiphinema parasetariae Luc. AB - Paralongidorus bullatus n. sp. from groundnut soils in Sador'e Niger is described and illustrated. It is distinguishable from most species of the genus by its body and odontostyle lengths, knob-like head, and digitate ending of the protoplasmic region of the tail. Length of the female body is 4.4-5.5 mm, of the odontostyle 132-156 mum, and of the tail 32-44 mum. Tail terminus is conoid to broadly rounded. Uteri are well developed, without sperms. Males are not found. Xiphinema parasetariae Luc 1958, a species inquirenda, is validated and, based on measurements and tail structures, X. attorodorum Luc 1961 is proposed as a synonym of X. parasetariae. Additional measurements are given for its females and juveniles. PMID- 19287762 TI - Resistance in Lycopersicon peruvianum to Isolates of Mi Gene-Compatible Meloidogyne Populations. AB - Root-knot nematode resistance of F progeny of an intraspecific hybrid (Lycopersicon peruvianum var. glandulosum Acc. No. 126443 x L. peruvianum Acc. No. 270435), L. esculentum cv. Piersol (possessing resistance gene Mi), and L. esculentum cv. St. Pierre (susceptible) was compared. Resistance to 1) isolates of two Meloidogyne incognita populations artificially selected for parasitism on tomato plants possessing the Mi gene, 2) the wild type parent populations, 3) four naturally occurring resistance (Mi gene)-breaking populations of M. incognita, M. arenaria, and two undesignated Meloidogyne spp., and 4) a population of M. hapla was indexed by numbers of egg masses produced on root systems in a greenhouse experiment. Artificially selected M. incognita isolates reproduced abundantly on Piersol, but not (P = 0.01) on resistant F hybrids. Thus, the gene(s) for resistance in the F hybrid differs from the Mi gene in Piersol. Four naturally occurring resistance-breaking populations reproduced extensively on Piersol and on the F hybrid, demonstrating ability to circumvent both types of resistance. Meloidogyne hapla reproduced on F hybrid plants, but at significantly (P = 0.01) lower levels than on Piersol. PMID- 19287763 TI - Efficient Procedure for Extracting Tylenchulus semipenetrans from Citrus Roots. AB - Investigations were undertaken to determine the suitability of sucrose and magnesium sulphate solutions and a silica colloidal suspension with centrifugation for extracting Tylenchulus semipenetrans from citrus roots. The efficiency of incubation, sodium hypochlorite, centrifugation, and maceration methods was also compared. Numbers of females recovered by centrifugation with colloidal silica were greater than those from sucrose or magnesium sulphate. Incubation, sodium hypochlorite, and centrifugation methods were satisfactory for extracting eggs, second-stage juveniles, and males, whereas the maceration sieving method was less efficient. Combining the sodium hypochlorite method with a 15-second maceration followed by centrifugation in colloidal silica reduced the recovery of T. semipenetrans females from citrus roots. PMID- 19287764 TI - Fungal Parasites of the Potato Cyst Nematode Globodera rostochiensis: Isolation and Reinfection. AB - Fungal parasitism of eggs of the potato cyst nematode Globodera rostochiensis was < 1, 3, and 17% at three sites in Sweden. The fungi isolated most frequently from infected eggs were a Septocylindrium-like fungus ( 19 %), Exophiala spp. (17 %), and Cylindrocarpon spp. (13 %). Verticillium suchtasporium was isolated from infected eggs at a low frequency (4%). In laboratory experiments V. suchlasporium infected 93% of the eggs within cysts after 10 days on dilute corn meal agar. This species showed chitinase and protease activity. Infection of eggs by the Septocylindrium-like fungus was moderate, whereas Cylindrocarpon destructans and Cladosporium cladosporoides did not infect eggs. No chitinase activity was found in these fungi, but protease activity was recorded in all. Growth of the fungi in cysts did not influence the number of physiologically disordered eggs. PMID- 19287765 TI - Ecdysteroids in Axenically Propagated Caenorhabditis elegans and Culture Medium. AB - Ecdysteroids (insect molting hormones) from Caenorhabditis elegans were chromatographically purified and quantified by radioimmunoassay. Nematodes from semidefined medium contained the immunoreactive equivalent of 460 pg ecdysone per gram dry weight. Culture medium, however, contained the immunoreactive equivalent of 68 times the quantity within the nematodes. In a defined medium lacking immunoreactivity, C. elegans contained 520 pg ecdysone equivalents per gram dry weight but reproduced slowly. Reproduction of C. elegans in defined medium was enhanced by formulation in agar. Propagation of C. elegans in either agar-based or aqueous defined medium supplemented with [(1)C]cholesterol of high specific activity failed to result in production of radiolabeled free ecdysteroids or polar or apolar ecdysteroid conjugates. Failure to demonstrate ecdysteroid biosynthesis in C. elegans raises questions about the ecdysteroids identified previously in nematodes being products of endogenous biosynthesis, a necessary condition for these compounds to be nematode hormones. PMID- 19287766 TI - Monoxenic Culture of Banana-Parasitic Nematodes on Musa acuminata cv. Poyo shoots. PMID- 19287768 TI - Differences in Egress of Male and Female Pratylenchus penetrans from Pea Roots. PMID- 19287767 TI - Influence of Potassium on Spore Germination in the Nematophagous Fungus, Hirsutella rhossiliensis. PMID- 19287769 TI - Heterodera glycines Invasion and Reproduction on Soybean Grown in Clay and Silt Loam Soils. PMID- 19287770 TI - Erratum. AB - [This corrects the article on p. 262 in vol. 22.]. PMID- 19287771 TI - An Assessment of Progress toward Microbial Control of Plant-Parasitic Nematodes. PMID- 19287772 TI - Soil fumigation: principles and application technology. AB - The principal soil fumigants and their order of discovery are carbon disulfide, chloropicrin, methyl bromide, 1,3-dichloropropene, ethylene dibromide, 1,2 dibromo-3-chloropropane, and methyl isothiocyanate. Biological activity of soil fumigants ranges from limited to broad spectrum. Fumigants diffuse through the continuous soil air space as gases. Physical and chemical characteristics determine diffusion rates, distribution between the soil air and moisture, and sorption onto and into the soil particles. The principal soil factors affecting the efficacy of each treatment are the size and continuity of air space, moisture, temperature, organic matter, and depth of placement. Application can be made overall with tractor injection or plow-sole, or as a row or bed treatment. Treatment for trees is best made in conjunction with tree site backhoeing. PMID- 19287773 TI - Spring or Fall Fumigation for Control of Meloidogyne spp. on Tobacco. AB - Tests were conducted in 1987-88 to compare the efficacy of spring or fall fumigant nematicide applications for control ofMeloidogyne arenaria and M. incognita on tobacco. Chloropicrin, 1,3-D, methyl isothiocyanate, and a methyl isothiocyanate-l,3-D mixture were applied as row treatments. Fenamiphos, fenamiphos + fensulfothion, or ethoprop were applied in the spring as nonfumigant nematicide standards. Fumigant nematicides increased yields and reduced galling (P = 0.01) in all four tests. Spring or fall applications of fumigant nematicides were effective in controlling M. arenaria and M. incognita and were superior to the nonfumigant nematicides tested. PMID- 19287774 TI - Reproduction of Meloidogyne incognita on Open-pollinated Maize Varieties. AB - Forty-three open-pollinated maize varieties were tested for resistance to the southern root-knot nematode, Meloidogyne incognita race 4, in greenhouse tests. An experiment repeated on five different planting dates assessed nematode reproduction 60 days after inoculation with 3,000 eggs per plant. Tebeau and Old Raccoon showed consistently high levels of resistance in all plantings, with the lowest reproduction factor (RF) values (0.2 and 0.4) and low numbers of eggs per gram of fresh root (222 and 955). Bill Dailey Variety and Sheppard Corn had the same level of resistance to M. incognita as the resistant hybrid, Mp307 x Mp707, but they were less consistent in different plantings. Levi Mallard, African Bushman, and Field's White Variety were the most susceptible varieties with RF values of 3.1-3.3 and 2,479-3,678 eggs per gram of fresh root. PMID- 19287775 TI - Effect of Oxamyl on Globodera tabacum Population Dynamics and Shade Tobacco Growth and Yield. AB - Preplant soil applications of oxamyl to shade grown tobacco in Globodera tabacum infested field soil increased green leaf yields over untreated plots by 10.7 and 21.0% for 2.2 and 6.7 kg a.i. oxamyl/ha, respectively. Green leaf yield was negatively correlated (r = -0.60, P = 0.04) with initial G. tabacum density, which ranged from 33 to 154 second-stage juveniles (J2)/cm(3) soil. Numbers of G. tabacum J2 and developing juveniles and adults (J3-adults) per gram root were fewer in plants from oxamyl-treated plots than in plants from untreated plots. Numbers of J2 in roots 4, 6, and 8 weeks after transplanting were reduced by 80, 89, and 4%, respectively, and numbers of J3-adults were reduced by 96, 89, and 21%, respectively, in high-rate oxamyl plots, compared with untreated plots. Globodera tabacum reproduction, as measured by the ratio of final to initial soil densities, was less in oxamyl-treated plots than in untreated plots. PMID- 19287776 TI - Effects of soil solarization on nematodes parasitic to chickpea and pigeonpea. AB - Solarization by covering the soil with transparent polyethylene sheets during the summer months (April, May, June) in 1984 and 1985 significantly (P = 0.01) reduced the population densities of nematodes (Heterodera cajani, Rotylenchulus reniformis, Helicotylenchus retusus, Pratylenchus sp., and Tylenchorhynchus sp.) parasitic to chickpea and pigeonpea. Population density reductions of 93% of Heterodera cajani eggs and juveniles, 99% ofHelicotylenchus retusus, 98% of Pratylenchus sp., and 100% of R. reniformis were achieved by solarization in 1984. Irrigation before covering soil with polyethylene improved (P = 0.01) the effects of solarization in reducing the population densities of Heterodera cajani. Similar trends in population density reductions were observed in 1985, but the solarization effects were not the same. Nematode population reductions in the 1984-85 season were evident until near crop harvest, but in the 1985-86 season the effects on nematode populations were not as great and did not last until harvest. Factors such as rains during the solarization, duration of solarization, and sunshine hours may have influenced the efficacy of solarization. Solarization for two seasons reduced the population densities each year about the same as single season solarization, and residual effects of solarization on nematode populations did not last for more than a crop season. PMID- 19287777 TI - Effects of soil fumigants and aldicarb on corky ringspot disease and trichodorid nematodes in potato. AB - In 1982 and 1983 the soil fumigants ethylene dibromide (EDB), EDB + chloropicrin, and 1,3-dichloropropene (1,3-D) applied with one or three chisels per row were evaluated singly and in combination with aldicarb on potato, Solanum tuberosum cvs. Atlantic and Sebago, for control of trichodorid nematodes and potato corky ringspot disease (CRS). In 1982 dosages per chisel for EDB, EDB + chloropicrin, and 1,3-D were 16.8, 23.9, and 56.1 liters/ha, respectively. EDB was applied at 12.6 liters per chisel per ha in 1983. Aldicarb was applied at 3.4 kg a.i./ha in the-row. Differences (P 30 on Cordell, which derives SCN resistance from PI 90763. The index was < 10 on PI 88788 for seven populations, but two populations had an index > 40 on Bedford, which derives resistance from PI 88788. When recommending cultivars to be planted by soybean producers, SCN reproduction should be determined on the cultivars, instead of on race differentials. PMID- 19287779 TI - Plant-parasitic nematodes and fungi associated with root rot of peas on prince edward island. AB - Eight commercial pea fields on Prince Edward Island were sampled in June and July over a 2-year period (1986-87) to determine soil population densities and the incidence of nematodes and fungi associated with root rot of peas. Root lesion nematodes (Pratylenchus spp.) were the dominant endoparasitic nematodes recovered from roots and soil. Low populations of the northern root-knot nematode (Meloidogyne hapla) were also present. Tylenchorhynchus spp. and Paratylenchus spp. were recovered frequently from soil in the root zone, and Helicotylenchus spp. were also frequent, but in low numbers. Fusarium solani was the most common fungal species isolated from the epicotyl and hypocotyl tissues of pea. Fusarium oxysporum was also isolated frequently, and both Fusarium species were found in soil from all fields. Rhizoctonia solani and Verticillium albo-atrum were common in hypocotyl tissue, but V. dahliae was isolated infrequently. Root rot was rated as severe in all fields and was positively and significantly correlated (P or= 10(6) CFU/mL count in the tracheal aspirate) criteria. Four hundred and seventy four (47.7%) patients were submitted to mechanical ventilation with 141 (29.7%) VAPs, with S. aureus as the most frequent agent (41.2%). The phenotype ORSA accounted for 47.5% and OSSA for 52.5%, predominant in late-onset VAPs with frequencies of 93.1% and 68.7%, respectively. Age > 60, use of corticoid and previous antibiotic therapy were related (p<0.05) with the development of VAP by ORSA. Mortality rate was higher (p>0.05) in the group with VAP by ORSA (37.9%). S. aureus was the main agent of VAPs, around half by ORSA, associated with age, late-onset VAP development and previous use of antibiotics and corticoids, but with no significant difference in mortality compared with VAP by OSSA. PMID- 19287839 TI - Prevalence and risk factors for Staphylococcus aureus in health care workers at a university hospital of Recife-PE. AB - Staphylococcus aureus is the main human pathogen that colonizes individuals in general population. The objective of the study was evaluate the epidemiological and sensitivity profile of S. aureus lineage, isolated in health care workers (HCW) of a University Hospital in Pernambuco state, Brazil. Biological samples of hands and nasal cavities were sown in agar sheep blood. Colonies under suspicion of being S. aureus were identified using Gram staining, catalase test and coagulase, mannitol-salty agar fermentation and DNAse agar. The resistance to mupirocin was analyzed through the Kirby Bauer technique. In relation to methicillin and vancomycin the determination was by the minimum inhibitory concentration method (E-test). From the 202 HCW evaluated, 52 were colonized by S. aureus (25,7%). The factors associated to the colonization by S. aureus were: age-group, professional category, use of individual protection equipments (frequency and numbers). All S. aureus isolate lineages were sensitive to mupirocin and vancomycin, and three of them were identified as methicillin resistant. The prevalence of MSSA and MRSA among HCW was considered low and was below the results described in the literature. The isolate S. aureus lineages have shown low resistance profile. PMID- 19287840 TI - Pseudomonas aeruginosa infections: factors relating to mortality with emphasis on resistance pattern and antimicrobial treatment. AB - A retrospective case-control study was conducted to investigate the risk factors for death among intensive care unit patients with Pseudomonas aeruginosa infection. Out of 131 patients investigated, 67 (51.1%) died within 30 days of being diagnosed with this infection. The mean duration of hospital stay before this diagnosis was 28.5 +/- 26.5 days. No association was found between bacterial resistance and death in this study (multiresistant p= 0.26; panresistant p= 0.42), but the adequacy of the initial treatment was inversely proportional to the degree of resistance. There was a tendency towards greater mortality among patients who received combination therapy (empirical p= 0.09; definitive p= 0.08), despite the greater frequency of appropriate treatment in these patients and the similar degree of severity in the two groups. This finding may be explained by pharmacodynamic parameters that were not studied or by the extensive use of aminoglycosides in the combination therapy, which play a controversial role in combination therapy due to their potential for renal toxicity. The multivariate analysis in our study demonstrated that age [odds ratio (OR) 1.04], septic shock (OR 15.4) and hypoalbuminemia (OR 0.32) were independent risk factors for death. PMID- 19287841 TI - Phenotypic detection and occurrence of extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli at a tertiary hospital in Trinidad & Tobago. AB - The incidence and distribution of ESBL producing microorganisms such as E. coli and K. pneumoniae have been demonstrated and varies in different health care facilities and as well as other countries This study was carried out to determine the frequency of occurrence and the antimicrobial susceptibility pattern of ESBL producing E. coli and K. pneumoniae species from clinical isolates at a tertiary hospital in Trinidad & Tobago. Standard microbiological procedures and automated MicroScan System was used to identify, screen for putative ESBL production and determine antimicrobial susceptibility of 1,118 clinical isolates of Enterobacteriaceae species at the microbiology laboratory of the Eric Williams Medical Science Complex, Trinidad & Tobago over a 36 months period. All ESBL producing isolates flagged by the automated system were further confirmed by E test method. The E-test confirmed a 15.2% ESBL rate among the K. pneumoniae isolates and 9.3% among the E. coli isolates. There was also a 1.8% rate of ESBL production in K. pneumoniae and 0.2% in E. coli isolates from specimens received from community health facilities into the laboratory. Isolates recovered from the intensive care unit of the hospital had 2.1% E. coli and 8.2% K. pneumoniae ESBL producers. Although all ESBL positive isolates were completely susceptible to imipenem and meropenem; and all positive K. pneumoniae isolates were susceptible to amikacin, there was a low susceptibility of ESBL positive E. coli to the aminoglycosides. However, susceptibility of these ESBL producing isolates to the fluoroquinolones varied. There is a high rate of ESBL production among isolates of E. coli and K. pneumoniae at this hospital that is linked to the extensive inappropriate use of third generation cephalosporins in the country. Further molecular studies are needed to characterize the types of these ESBL prevailing in the country. PMID- 19287842 TI - Donovanosis. AB - Donovanosis is a chronic bacterial illness frequently associated with sexually transmitted infections (STI) and is under diagnosed both in endemic areas as well as in countries in which doctors have little experience with tropical diseases. The utilization of syndromic diagnosis and treatment of STIs in various parts of the world and the previous use of antibiotics make it difficult to find Donovan bodies in the cytodiagnostic and hystopathological exams, requiring the utilization of technology that is neither routine nor often accessible to confirm the hypothesized diagnosis. Therefore, it is necessary to bring medical professionals up to date about this infectious disease. PMID- 19287843 TI - Biofilms: microbes and disease. AB - Bacteria that attach to surface aggregate in a hydrated polymeric matrix of their own synthesis to form biofilms. These represent microbial societies with their own defense and communication system. Transitioning from acute to chronic infection is frequently associated with biofilm formation.Bacteria in biofilms are innately more resistant to antimicrobial agents. The presence of indwelling medical devices increases the risk for biofilm formation and subsequent infection. The current antibiotic therapies are of limited effectiveness in resolving biofilms infection.This review attempts to discuss the stages in biofilm formation, their pathogenic mechanisms, effect of antimicrobial agents, detection and eradication of the biofilms. PMID- 19287844 TI - Vascular access-related infections in HIV patients undergoing hemodialysis: case description and literature review. AB - Poor immune status, the use of a vascular access different from an AV fistula, and intravenous drug use (IDU) may favor increased rates of vascular access infections among HIV infected patients on hemodialysis. Staphylococcus spp. and Streptococcus spp. are the main cause of these infections, but Gram-negative rods and fungi have been found as well. Using an AV fistula when possible, and eliciting a history of IVDU on every visit may prevent this type of infection. When infections are present, coverage for both Gram-positive and negative organisms is recommended. Additional studies specifically addressing the issue of vascular access infection in HIV infected patients are required. PMID- 19287845 TI - Association of HTLV-I with Arnold Chiari syndrome and syringomyelia. AB - HTLV-I is associated with a broad spectrum of manifestations, including tropical spastic paraparesis and adult T-cell leukemia/lymphoma. Arnold Chiari syndrome is a condition characterized by herniation of the cerebellar tonsils through the foramen magnum. This condition should be suspected in all patients with headache and impaired motor coordination. Syringomyelia is a developmental anomaly that leads to the formation of an intramedullary cavity. Its clinical presentation is classically characterized by syringomyelic dissociation of sensation, with suspended distribution in the proximal portion of the trunk and upper limbs and preservation in other regions. We report here a case of association of the three diseases, which is rare in clinical practice, illustrating the difficulty in the diagnosis and therapeutic management of these conditions. PMID- 19287846 TI - Spontaneous splenic rupture due to dengue fever: report of two cases. AB - Dengue is a febrile illness caused by Flavivirus and mainly transmitted by the mosquito Aedes aegypiti which have been a serious epidemic in Rio de Janeiro. In most of cases it was a self limited disease. We report two cases of a serious and rare complication of this viral infection. PMID- 19287847 TI - First reported case of infective endocarditis caused by community-acquired methicillin-resistant Staphylococcus aureus not associated with healthcare contact in Brazil. AB - We report here the first case of endocarditis due to CA-MRSA not associated with healthcare contact in Brazil in Brazil. A previously healthy patient presented with history of endocarditis following a traumatic wound infection. Patient had multiple positive blood cultures within 72 h of admission and met modified Duke's criterion for infective endocarditis. The isolate was typed as Staphylococcal cassette chromosome (SCC) mec type IV and was positive for presence of Panton Valentine leukocidin (PVL). Increased incidence of CA-MRSA endocarditis is a challenge for the internist to choose the best empirical therapy. Several authors have suggested an empirical therapy with both a beta-lactam and an anti-MRSA agent for serious S. aureus infections. Our patient was treated with Vancomycin and made complete recovery in 3 months. PMID- 19287848 TI - Spondylodiscitis and infectious endocarditis: a round-trip to be considered. AB - The association between spondylodiscitis and endocarditis was first reported in 1965 by de Seze et al. The most common clinical picture of this association is musculoskeletal symptoms preceding endocarditis diagnosis, but we report here a case of spondylodiscitis complicating endocarditis in its late course. A 70-year old man, with an established diagnosis of mitral valve endocarditis caused by Streptococcus intermedius, early submitted to surgical treatment because of heart failure, who had an uneventful recovery up to the 12th day of antibiotic therapy when he presented intensive backache, with tenderness in the two lower lumbar vertebras. Spondylodiscitis was confirmed by a magnetic resonance imaging and the treatment was non-esteroidal anti-inflammatory and analgetics drug, with good results, and prolongation of antibiotic treatment up to 3 months. Appropriate diagnosis of this association has important consequences, as the need of a longer antibiotic therapy course, which can range from 6 weeks to 3 months. PMID- 19287849 TI - Intrathecal use of amikacin: a case report. AB - Meningitis caused by Acinetobacter baumannii is rare and are mostly hospital acquired after neurosurgical procedure. We report a case of a 40-year old man was admitted to the intensive care unit due to subarachnoid haemorrhage. Our patient developed a ventriculitis due to A.baumannii treated successfully with sulbactam IV and intrathecal amikacin. PMID- 19287850 TI - Aeromonas caviae septicemia in immunocompetent gastrointestinal carriers. AB - Aeromonas caviae strains have been isolated from blood and stool cultures of three immunocompetent patients, residents of Northern India, who presented with community acquired septicemia without any recent history of diarrhea. Cell culture infectivity test performed on Hep-2 cells have shown substantial degree of invasiveness in the isolated strains. This case unleashes a possibility of asymptomatic gastrointestinal carriage of such strains of A. caviae in a very large population of India, as several areas of India have very high rates of Aeromonas induced acute diarrhea/gastroenteritis (up to 13%). It needs to be appraised further in India as well as other countries having high rates of Aeromonas induced acute diarrhea/gastroenteritis. PMID- 19287851 TI - Cases of OPSI syndrome still candidate for medical ICU. AB - Splenectomized patients are likely to suffer from severe infections, such as sepsis and meningitis, which is called overwhelming postsplenectomy infection (OPSI) syndrome. It seems to be more common in children, but occurs at all ages. The risk is greatest in the early months and years after operation, but never disappears entirely. The course is rapid, the clinical symptoms are serious, and the prognosis is very poor. In this paper, three cases of OPSI syndrome are described, in which infection developed 8, 8 and 15 years after splenectomy; two of the patients died. With the help of these case reports, we want to again emphasize the importance of vaccination, antibiotic prophylaxis and seeking earlier medical attention in splenectomized patients. PMID- 19287852 TI - Emphysematous cystitis--a case report. AB - Emphysematous Cystitis is a primary infection of the bladder with production of gas by bacteria. The infection is uncommon, still has obvious clinical importance due to its morbidity and mortality potential, as the following case enlightens. We report a clinical case of a patient admitted with acute myocardial infarction who developed an acute emphysematous cystitis, a further complication in his long and complex period of hospitalization. PMID- 19287853 TI - Chronic meningitis by histoplasmosis: report of a child with acute myeloid leukemia. AB - Meningitis is a common evolution in progressive disseminated histoplasmosis in children, and is asymptomatic in many cases. In leukemia, the impaired of the T cells function can predispose to the disseminated form. The attributed mortality rate in this case is 20%-40% and the relapse rate is as high as 50%; therefore, prolonged treatment may be emphasized. We have described a child with acute myeloid leukemia (AML), that developed skin lesions and asymptomatic chronic meningitis, with a good evolution after prolonged treatment with amphotericin B deoxycholate followed by fluconazole. PMID- 19287854 TI - Usefulness of hybrid SPECT/CT for the 99mTc-HMPAO-labeled leukocyte scintigraphy in a case of cranial osteomyelitis. AB - Cranial osteomyelitis is a potentially fatal lesion. White blood cell scanning (WBC) with 99mTc-hexamethylpropylene amine oxime (HMPAO) has proven highly sensitive and specific in the diagnosis and follow-up of patients with suspected osteomyelitis. In this report we show the usefulness of SPECT and transmission CT performed simultaneously using a hybrid imaging device for the functional anatomic mapping of soft tissue and cranial bone infections. 99mTc-HMPAO-labeled leukocytes scintigraphy was performed on an elderly diabetic man with an intracranial mass lesion and with suspected temporal bone infection. Planar scans were acquired 30 min, 4 h, and 24 h after injection. SPECT/CT was obtained 6 h after tracer injection, using a dual-head camera coupled with a low-power X-ray tube. The scintigraphic results were matched with the results of surgery and of clinical follow-up. The planar images alone were true-positives for abscess in this patient. SPECT/CT improves the accuracy of99mTc-HMPAO scintigraphy especially in discriminating between soft-tissue and bone involvement. In fact, SPECT/CT also showed temporal bone osteomyelitis. This result indicates that SPECT/CT performed using a hybrid device can improve imaging with 99mTc-HMPAO labeled leukocytes in patients with suspected osteomyelitis by providing accurate anatomic localization and precise definition of the extent of infection. PMID- 19287855 TI - An ecosystem perspective in the socio-cultural evaluation of dengue in two Colombian towns. AB - Despite extensive public health efforts, dengue is still a major health concern in Colombia. The objective of this study was to provide an ecosystem and cross disciplinary perspective on the dengue situation in two Colombian towns. The article focuses on presenting the anthropological methodology and research findings. An interdisciplinary team gathered quantitative (cross-sectional), meteorological, entomological, and qualitative data (based on medical anthropology) through fieldwork and archival research. According to the qualitative data, dengue can be described as a point of convergence between public health policies, the affected population, the environment, and the social dynamics generated through this interaction. Dengue is illustrative as a disease, in that it has a negative impact on public health, but individuals in Colombia have learned to live and cope with it. Dengue prevalence and its on-going historical presence have made it part of everyday community life, viewed as a minor health issue. PMID- 19287856 TI - Dengue in Southeast Asia: epidemiological characteristics and strategic challenges in disease prevention. AB - Dengue emerged as a public health burden in Southeast Asia during and following the Second World War and has become increasingly important, with progressively longer and more frequent cyclical epidemics of dengue fever/dengue hemorrhagic fever. Despite this trend, surveillance for this vector-borne viral disease remains largely passive in most Southeast Asian countries, without adequate laboratory support. We review here the factors that may have contributed to the changing epidemiology of dengue in Southeast Asia as well as challenges of disease prevention. We also discuss a regional approach to active dengue virus surveillance, focusing on urban areas where the viruses are maintained, which may be a solution to limited financial resources since most of the countries in the region have developing economies. A regional approach would also result in a greater likelihood of success in disease prevention since the large volume of human travel is a major factor contributing to the geographical spread of dengue viruses. PMID- 19287857 TI - Irregular water supply, household usage and dengue: a bio-social study in the Brazilian Northeast. AB - Despite increased vector control efforts, dengue fever remains endemic in Fortaleza, Northeast Brazil, where sporadic epidemic outbreaks have occurred since 1986. Multiple factors affect vector ecology such as social policy, migration, urbanization, city water supply, garbage disposal and housing conditions, as well as community level understanding of the disease and related practices. This descriptive study used a multi-disciplinary approach that bridged anthropology and entomology. A multiple case study design was adopted to include research in six study areas, defined as blocks. The water supply is irregular in households from both under-privileged and privileged areas, however, clear differences exist. In the more privileged blocks, several homes are not connected to the public water system, but have a well and pump system and therefore irregularity of supply does not affect them. In households from under-privileged blocks, where the water supply is irregular, the frequent use of water containers such as water tanks, cisterns, barrels and pots, creates environmental conditions with a greater number of breeding areas. In under-privileged homes, there are more possible breeding areas and environmental conditions that may improve the chances of Aedes aegypti survival. PMID- 19287858 TI - [Information, knowledge, and perception of dengue risk in Argentina: two intervention experiments to generate local control strategies]. AB - With the objectives of dengue prevention, health promotion, and action-based research, two experiments were conducted in pilot areas of Buenos Aires and Vicente Lopez, Argentina. In each area, community, government, and non governmental references were identified. The study included an entomological survey (with ovitraps), environmental survey (with field observations, interviews, and workshops), and social survey (using questionnaires, interviews, and meetings). Spaces for dialogue and collaboration were developed with the community, thereby spawning participatory activities and empowerment. Environmental reordering and reduction of mosquito breeding sites were promoted by: incorporation of environmental issues and vector-borne disease prevention into the school curricula; neighborhood workshops; training of environmental agents to transmit information and train peers; and planning community environmental projects. The households were visited, seeking to integrate the residents into their own community environmental diagnosis. There were significant differences between the pilot areas in knowledge and social practices, but not according to socioeconomic status. The schools and health promotion settings proved to be the main community reference for promoting healthy environmental practices. PMID- 19287859 TI - Health and sustainable development: challenges and opportunities of ecosystem approaches in the prevention and control of dengue and Chagas disease. AB - A world of healthy people living in healthy ecosystems has proven to be an elusive goal of the sustainable development agenda. Numerous science-based assessments agree on the fundamental interdependence between people's health, the economy, and the environment, and on the urgency for more determined and concerted action based on multi-sector participatory approaches at the global and local levels. For knowledge to be policy-relevant and capable of contributing to healthy and sustainable development, it must take into account the dynamic and complex interactions between ecological and social systems (systems thinking), and it must be linked to development actions. This in turn requires greater interaction and exchange between decision-makers, researchers and civil society (a multi-stakeholder participatory process); and the harnessing of different disciplines and of different kinds of knowledge (a transdisciplinary approach). Ecosystem approaches to human health (ecohealth) link these elements in an adaptable framework for research and action. This paper presents an overview of ecohealth research approaches applied to vector-borne diseases, with particular attention to multi-stakeholder participation given its prominence in the sustainable development policy discourse. PMID- 19287860 TI - The ecological dimensions of vector-borne disease research and control. AB - Alarming trends in the resurgence of vector-borne diseases are anticipated to continue unless more effective action is taken to address the variety of underlying causes. Social factors, anthropogenic environmental modifications and/or ecological changes appear to be the primary drivers. The ecological dimension of vector-borne disease research and management is a pervasive element because this issue is essentially an ecological problem with biophysical, social, and economic dimensions. However there is often a lack of clarity about the ecological dimension, the field of ecology (e.g. role, limitations), and related concepts pertinent to ecosystem approaches to health. An ecological perspective can provide foresight into the appropriateness of interventions, provide answers to unexpected vector control responses, and contribute to effective management solutions in an ever-changing environment. The aim of this paper is to explore the ecological dimension of vector-borne diseases and to provide further clarity about the role of 'ecological thinking' in the development and implementation of vector control activities (i.e. ecosystem approaches to vector-borne diseases). PMID- 19287861 TI - House improvements and community participation in the control of Triatoma dimidiata re-infestation in Jutiapa, Guatemala. AB - The deterioration or absence of plaster walls in houses and poor hygienic conditions are the most important risk factors for indoor Triatoma dimidiata infestation in Guatemala. A cross-disciplinary study was conducted addressing T. dimidiata infestation, household hygiene, and housing construction. The study focused on local materials and cultural aspects (including gender roles) that could lead to long-term improvements in wall construction. A new plaster mix for walls was developed on the basis of laboratory studies on construction materials recommended by local villagers. Four villages with persistent (post-spraying) T. dimidiata infestation were studied. In two villages, an ecosystem approach was implemented, and the homeowners conducted wall improvements and household sanitation with the support of the interdisciplinary team (the ecosystem intervention). In the other two villages, a vector control approach based on insecticide spraying was adopted (traditional intervention). Both interventions were associated with a reduction in T. dimidiata infestation, but only the ecosystem approach produced important housing improvements (sanitation and wall construction) capable of preventing T. dimidiata re-infestation in the long term. PMID- 19287862 TI - [Chagas disease in a life story: unveiling the disease, changing the world. Interview with Joao Carlos Pinto Dias. Interview by Roberto Briceno-Leon]. PMID- 19287863 TI - Dengue in the Americas: challenges for prevention and control. AB - Dengue is the most important vector-borne disease in the Americas and threatens the lifes of millions of people in developing countries. Imprecise morbidity and mortality statistics underestimate the magnitude of dengue as a regional health problem. As a result, it is considered a low priority by the health sector with no timely steps for effective control. Dengue is perceived as a problem of 'others' (individually, collectively and institutionally), therefore responsibility for its control is passed on to others (neighbors, the community, municipality, health institutions, or other governmental agencies). With no precise risk indicators available there is little opportunity for timely diagnoses, treatment, health interventions or vector control (poor surveillance). Solutions only targeting the vector reduce the impact of interventions and there is no sustainable control. Without political commitment there are insufficient resources to face the problem. This paper discusses the challenges for prevention and control in the Americas. PMID- 19287864 TI - Environment, interactions between Trypanosoma cruzi and its host, and health. AB - An epidemiological chain involving Trypanosoma cruzi is discussed at the environmental level, and in terms of fine molecular interactions in invertebrate and vertebrate hosts dwelling in different ecosystems. This protozoan has a complex, genetically controlled plasticity, which confers adaptation to approximately 40 blood-sucking triatomine species and to over 1,000 mammalian species, fulfilling diverse metabolic requirements in its complex life-cycle. The Tr. cruzi infections are deeply embedded in countless ecotypes, where they are difficult to defeat using the control methods that are currently available. Many more field and laboratory studies are required to obtain data and information that may be used for the control and prevention of Tr. cruzi infections and their various disease manifestations. Emphasis should be placed on those sensitive interactions at cellular and environmental levels that could become selected targets for disease prevention. In the short term, new technologies for social mobilization should be used by people and organizations working for justice and equality through health information and promotion. A mass media directed program could deliver education, information and communication to protect the inhabitants at risk of contracting Tr. cruzi infections. PMID- 19287865 TI - Ecosystem approaches to controlling of vector-borne diseases: dengue and Chagas disease. PMID- 19287866 TI - Preliminary evaluation of the 'Dengue-MI' technology for Aedes aegypti monitoring and control. AB - Limitations in the laboratory identification of Aedes aegypti and processing of field data based on larval surveys led to the development of the 'Intelligent Dengue Monitoring' technology (MI-Dengue). MI-Dengue consists of a trap that captures gravid female Ae. aegypti, coupled with a computerized system for field data collection, transmission, and access to georeferenced maps in real time. The current study describe the first experience with a system for monitoring adult Ae. aegypti and presents the preliminary results in three municipalities that adopted MI-Dengue as a strategy to identify key areas and orient control measures. Weekly georeferenced maps and an entomological indicator (Mean Female Aedes Index) provided information on infested areas and infestation levels, color coded according to the number of captured female Ae. aegypti, and indicated risk free, dengue alert, and critical situations that triggered appropriate control measures. The preliminary results suggest that the adoption of this control strategy with house-to-house visits in a 200m radius of the positive trap helped reduce dengue in the municipalities that adopted the system. PMID- 19287867 TI - [An inter-sector participatory strategy in Cuba using an ecosystem approach to prevent dengue transmission at the local level]. AB - Cuba is located among a group of countries with high dengue incidence. Following several epidemics in the last 10 years, the country designed, implemented, and evaluated a participatory strategy based on the Ecohealth approach. The aim was to promote inter-sector ecosystem management to decrease Aedes aegypti infestation and prevent dengue transmission in the municipality of Cotorro, in Havana city. The study adopted a participatory research methodology. The strategy ensured active participation by the community, diverse sectors, and government in the production of healthy ecosystems. Timely and integrated measures for prevention and control were developed, thereby decreasing the risk of vector proliferation and local dengue transmission. The approach allowed holistic problem analysis, priority setting, and administration of solutions. The strategy has been sustained two years after concluding the process. PMID- 19287868 TI - Dengue: twenty-five years since reemergence in Brazil. AB - This article revisits the epidemiology of dengue in Brazil, 25 years after its reemergence in the country, discussing the main determinants and implications for its control. The authors emphasize the clinical and epidemiological peculiarities of this viral disease in Brazil, which leads the world in both the number of cases reported and risk of occurrence of the disease. The article presents the changes occurring in the dengue epidemiological pattern in recent years, like the sudden age shift in the incidence of dengue hemorrhagic fever, and discusses possible associated factors. The article focuses specifically on the epidemic in Rio de Janeiro in 2008, this episode's impact on the international community, and the fear that the disease could spread to Europe. The authors conclude that the current situation emphasizes the need for the international scientific community to renew its efforts to generate knowledge allowing improvement and progress in the development of new tools and strategies for dengue prevention. PMID- 19287869 TI - [Chagas disease in the Americas: an ecohealth perspective]. AB - The historical processes involved in Chagas disease transmission relate to the patterns and conditions of human settlements, especially in rural areas, due to proximity to forest areas, where both vectors and Trypanosoma cruzi can occur, combined with precarious housing conditions and underlying poverty. However, seasonal and permanent rural-urban migration has played a major role in re mobilizing vectors, T. cruzi, and Chagas-infected individuals. A new agricultural frontier in the Amazon has led to a new transmission pattern, especially with palm trees located close to houses. Improved blood bank surveillance has decreased transmission by blood transfusions. International migration also plays a role in Chagas disease epidemiology. The United States and Spain, where specific health services for Chagas disease diagnosis and treatment are largely absent, harbor an unknown number of individuals with Chagas, probably infected decades ago. The article discusses major strides in Chagas disease knowledge and control, besides identifying persistent gaps, such as the need for housing improvements, especially in poor rural areas in the Americas. PMID- 19287870 TI - Risk factors for intradomiciliary infestation by the Chagas disease vector Triatoma dimidiatain Jutiapa, Guatemala. AB - Seventeen variables were evaluated as possible risk factors for the intradomiciliary infestation with Triatoma dimidiata in 644 houses in Jutiapa, Guatemala. During 2004 the houses were assessed for vector presence and evaluated for hygiene, cluttering, material comfort, construction conditions and number of inhabitants, among other factors. Chi-square analysis detected significant associations between vector presence and eight variables related to domestic sanitary and construction conditions. Log-linear models showed that regardless of the age of the house, the odds of vector presence were 4.3 and 10 times lower in houses with a good socioeconomic status compared with poor and very poor houses respectively. Log-linear models also pointed to a greater chance of vector presence when walls lacked plastering (3.85 times) or walls had low quality incomplete plastering (4.56 times), compared with walls that were completely plastered. Control strategies against T. dimidiata should include the introduction of better-quality but inexpensive plastering formulations and better sanitation practices should also be promoted among the population. Such control strategies should not only reduce or eliminate infestation, but also prevent vector reinfestation. PMID- 19287871 TI - An ecosystemic approach to evaluating ecological, socioeconomic and group dynamics affecting the prevalence of Aedes aegypti in two Colombian towns. AB - This article focuses on the epidemiological methods and results of a global Ecohealth study that explored the complexity of the relationship between ecological, biological, economical, social and political factors and vector presence. The study was carried out in two dengue endemic areas of Colombia. A transdisciplinary team gathered quantitative and qualitative data. A survey in randomly sampled households was applied and, simultaneously, direct observation of potential breeding sites was carried out. Logistic regressions and qualitative techniques were used. Qualitative and quantitative data were compared using triangulation. The presence of low water containers increases seven-fold the risk of finding immature forms of Aedes aegypti in the household (OR = 7.5; 95%CI: 1.7 32.2). An inverse association between socioeconomic stratum and presence of the vector was identified (Low stratum OR = 0.9; 95%CI: 0.6-1.4; High stratum OR =0.4; 95%CI: 0.07-1.7). Water management is a complex social dynamic associated with the presence of Ae. aegypti. Dengue control is a challenge for public health authorities and researchers as they should address promotion and prevention strategies that take into account cultural, behavioral, socioeconomic and health factors. PMID- 19287872 TI - [Predictive factors of disability in patients with leprosy]. AB - OBJECTIVE: To analyze predictive factors in the progression of the disability grade in patients with leprosy. METHODS: A retrospective cohort study followed up 595 patients with disability registered at a healthcare unit in the city of Belo Horizonte (Southeastern Brazil) from 1993 to 2003. Patients' sociodemographic and clinical information was collected from the respective medical records. Comparisons were made between the disability grade upon admission and at the end of treatment using a marginal homogeneity test. To determine factors associated with progression in the disability grade, univariate analysis (linear trend chi square test) was employed, as well as multivariate analysis by means of the algorithm Chi-square Automatic Interaction Detector. RESULTS: Among the cases in which the disability grade was recorded upon admission and upon discharge, 43.2% of the patients who had grade 1 in the first assessment progressed to grade 0. Among those who began with grade 2, 21.3% progressed to grade 0 and 20% progressed to grade 1. In the univariate analysis, the variables that proved to be statistically associated with progression in the disability grade were: neuritis, time elapsed until the occurrence of neuritis, number of damaged nerves, type of physiotherapy treatment and higher dose of prednisone. In the multivariate analysis, the main factor associated with the progression of disability was the disability grade upon admission. CONCLUSIONS: The results showed the importance of an early diagnosis of neuropathy as well as the efficient association of pharmacological and non-pharmacological treatment, through disability prevention techniques and adequate doses of steroid. PMID- 19287873 TI - [Land transport injuries among emergency department visits in the state of Sao Paulo, in 2005]. AB - OBJECTIVE: To analyze the characteristics of visits resulting from land transport injuries. METHODS: A total of 5,934 visits in four hospital emergency departments (ED) were analyzed, in the state of Sao Paulo, in 2005. A questionnaire based on the following three models was used to collect data: World Health Organization (WHO), Center for Disease Control and Prevention (CDC), and Pan American Health Organization (PAHO). Variables analyzed were as follows: type of road user (vehicle occupant, pedestrian, motorcyclist, and cyclist), sex, age group, and type of injury suffered. Logistic regression analysis was employed to test associations between variables. Odds ratios with their respective 95% confidence intervals were calculated. RESULTS: The majority of victims were males (74.2%) in the 20-to-29-year age group (35.0%). Vulnerable road users totaled 72.4% of all cases (29.8% were motorcyclists, 24.1% pedestrians, and 18.5% cyclists). Victims aged between zero and 14 years who had suffered injuries were mostly pedestrians and cyclists; motorcyclists predominated among those aged between 15 and 39 years; and pedestrians among those aged over 50 years. About half of the cases suffered minor injuries (strains, dislocations, contusions and cuts), while the other half was comprised by fractures, traumatic brain injuries and internal injuries. Extremities were the most affected body parts, particularly among motorcyclists. The majority of victims were discharged at triage (87.6%). Compared to women, men were 1.5 times more likely to be admitted or transferred, or to die. Pedestrians, vehicle occupants and motorcyclists were, respectively, 2.7, 2.4 and 1.9 times more likely to be admitted or transferred, or to die than cyclists. CONCLUSIONS: Measures aimed to protect vulnerable road users should be among the priorities to reduce land transport-related injuries. PMID- 19287874 TI - Factors related to inadequate cervical cancer screening in two Brazilian state capitals. AB - OBJECTIVE: To analyze factors associated with cervical cancer screening failure. METHODS: Population-based cross-sectional study with self-weighted two-stage cluster sampling conducted in the cities of Fortaleza (Northeastern Brazil) and Rio de Janeiro (Southeastern Brazil) in 2002. Subjects were women aged 25-59 years in the last three years prior to the study. Data were analyzed through Poisson regression using a hierarchical model. RESULTS: The proportion of women who did not undergo the Pap smear test in Fortaleza and Rio de Janeiro was 19.1% (95% CI: 16.1;22.1) and 16.5% (95% CI: 14.1;18.9), respectively. Higher prevalence ratios of cervical cancer screening failure in both cities were seen among women with low education and low per capita income, old age, unmarried, who never underwent mammography, clinical breast examination, and blood glucose and cholesterol level testing. Smokers also had lower screening rates compared to non smoker women and this difference was only statistically significant in Rio de Janeiro. CONCLUSIONS: The study findings point to the need of intervention focusing particularly women in worse socioeconomic conditions and access to healthcare, old-aged and unmarried. Education activities must prioritize screening of asymptomatic women and early diagnosis for symptomatic women and access to adequate diagnostic methods and treatment should be provided. PMID- 19287875 TI - Interactions between physical and psychosocial demands of work associated to low back pain. AB - OBJECTIVE: To examine the interaction between physical and psychosocial demands of work associated to low back pain. METHODS: Cross-sectional study carried out in a stratified proportional random sample of 577 plastic industry workers in the metropolitan area of the city of Salvador, Northeast Brazil in 2002. An anonymous standard questionnaire was administered in the workplace by trained interviewers. Physical demands at work were self-rated on a 6-point numeric scale, with anchors at each end of the scale. Factor analysis was carried out on 11 physical demand variables to identify underlying factors. Psychosocial work demands were measured by demand, control and social support questions. Multivariate analysis was performed using the likelihood ratio test. RESULTS: The factor analysis identified two physical work demand factors: material handling (factor 1) and repetitiveness (factor 2). The multiple logistic regression analysis showed that factor 1 was positively associated with low back pain (OR=2.35, 95% CI 1.50;3.66). No interaction was found between physical and psychosocial work demands but both were independently associated to low back pain. CONCLUSIONS: The study found independent effects of physical and psychosocial work demands on low back pain prevalence and emphasizes the importance of physical demands especially of material handling involving trunk bending forward and trunk rotation regardless of age, gender, and body fitness. PMID- 19287876 TI - [Level of noise at the workplace environment among physical education teachers in indoor bike classes]. AB - OBJECTIVE: To analyze the level of noise at the workplace environment among Physical Education teachers during indoor bike classes and its association with some aspects of health. METHODS: Cross-sectional study carried out in 15 different gyms with 15 teachers in Rio de Janeiro (Southeastern Brazil) in 2007. A standardized questionnaire, tested in relation to its replication, was used in order to find out the characteristics of the process and the way this job is organized, as well as complaints related to health reported by these teachers. The SRQ-20 (Self-Report Questionnaire) was utilized in order to verify the existence of minor psychiatric disturbances. The levels of sound pressure were measured by a portable appliance. The level of pressure was verified in dB(A) at equivalent energy levels (Leq) in different places of the room at different moments during the class. ANOVA, chi-square and Pearson's correlation analyses were performed. RESULTS: The levels of sound pressure varied from 74.4 dB(A) to 101.6 dB(A). The results during the class were: a) warm-up (mean=88.45 dB(A)); b) main part (mean= 95.86 dB(A)); and, closure (mean= 85.12 dB(A)). The mean background noise was 66.89 dB(A)). There were significant differences (p<0.001) among the background noise levels within different moments of the class. The noise was not related to minor psychiatric disturbances. CONCLUSIONS: Physical education professionals working with indoor bike classes are under high levels of sound pressure during their classes. This physical agent has been related to several health problems and should therefore be more controlled. PMID- 19287877 TI - Evidence of inadequate docosahexaenoic acid status in Brazilian pregnant and lactating women. AB - Recently published data concerning dietary intake of fat and food sources of (n 3) long-chain polyunsaturated fatty acids (LCPUFA) in Brazil are reviewed together with data on biochemical indices of PUFA status during pregnancy and lactation and PUFA composition of breast milk in Brazilian adolescents and adults. Potential inadequacies of docosahexaenoic acid (DHA) status among Brazilian pregnant and lactating women have not yet been thoroughly evaluated. The data reviewed show that dietary intake of food sources of n-3 LCPUFA is low and possibly deficient in Brazil, and that biochemical indices of maternal DHA status and breast milk DHA content are low compared to the international literature. These data indicate inadequate DHA status among Brazilian women during pregnancy and lactation, but this evidence needs confirmation through comprehensive and specific population-based studies. PMID- 19287878 TI - [Prevalence of auditory and vestibular symptoms among workers exposed to occupational noise]. AB - The purpose of the study was to assess the prevalence of auditory and vestibular symptoms in workers exposed to occupational noise. There were examined medical records of 175 workers with noise-induced hearing loss who attended an occupational health reference center in the city Campinas, Southeastern Brazil, from 1997 to 2003. The variables studied were frequency of symptoms of hypoacusis, tinnitus, and vertigo. Association with age, noise exposure time, and auditory thresholds were analyzed using the chi-square test and Fisher's exact test. Hypoacusis was reported in 74% of cases, tinnitus in 81%, and vertigo in 13.2%. There was found an association between hypoacusis and age, noise exposure time, and auditory thresholds and between vertigo and noise exposure time. No other significant associations were found. PMID- 19287879 TI - [Chagas disease centenary]. PMID- 19287880 TI - [Guidelines for Voluntary Counseling and Testing Centers (VCT) in the state of Sao Paulo, Brazil]. PMID- 19287881 TI - [Brazilian Registry of Clinical Trials (Rebrac): strengthening of clinical trials management in Brazil]. PMID- 19287883 TI - High dietary calcium intake decreases bone mobilization during pregnancy in humans. AB - Calcium metabolism of the mother is modified during pregnancy because of the mineralization of the fetus skeleton. OBJECTIVE: To evaluate the association of calcium intake and bone demineralization during pregnancy. MATERIAL AND METHODS: At each trimester of pregnancy a validated food frequency intake questionnaire was administered to assess individual daily calcium intake in a cohort of 206 pregnant women, residents of Mexico City. Samples of urine were collected to measure levels of the cross-linked N-telopeptide of type I collagen (NTx), which is a biomarker of bone resorption. The association between calcium ingestion and bone resorption was analyzed using random effects models; non-linear associations were explored using generalized additive models. RESULTS: Progressive increases in NTx levels were observed during pregnancy; with mean and standard deviation (SD) values during the first, second and third trimester of 76.50 (SD=38), 101.02 (SD=48.86) and 144.83 (SD=61.33) nmol BCE/mmol creatinine, respectively. Higher dietary calcium intake was associated with lower bone resorption (beta=-0.015; p<0.05). The association between age and NTx showed a non-linear trend with an inflexion point around 33 years: increase in maternal age below that point was associated with a decrease in bone resorption, while in older women the increase in age was associated with an increased resorption. CONCLUSIONS: Our results suggest that calcium ingestion, specifically from dairy products, reduces bone resorption during pregnancy. For each 300 mg (a glass of milk) of calcium intake there is an estimated reduction in NTx level of 4.8 nmol BCE/mmol of creatinine (p<0.05). PMID- 19287884 TI - Direct costs of osteoporosis and hip fracture: an analysis for the Mexican Social Insurance Health Care System. AB - OBJECTIVE: To compare costs of diagnosis and annual treatment of osteoporosis and hip fracture between the Instituto Nacional de Rehabilitacion (INR) and the protocol used by the Seguro Popular de Salud (SPSS). METHODS: Direct costs gathered in a prospective study with real cases at the INR are presented, and then this data is re-analyzed with the methodology and protocol for the SPSS to estimate the costs of those cases if treated with the SPSS protocol. RESULTS: Important differences were found in the cost of hip fracture: the SPSS estimates ($37,363.73 MXN) almost double the INR cost ($20,286.86 MXN ). This discrepancy was caused by the different types of surgeries the INR and SPSS protocols call for (the SPSS assumes that all hip fractures will necessitate a hip replacement) and the cost of subsequent hospitalization. A prospective study at the SPSS is needed to validate these results. CONCLUSIONS: Important differences were found between treatment of the same osteoporosis related problems at the INR and SPSS. We recommend revising the SPSS protocol to include less costly surgical treatments. PMID- 19287885 TI - Treatment options for osteoporosis and decision making criteria: 2009. AB - Osteoporosis is recognized worldwide as a major public health problem since many decades ago, mainly due to the cost of treatment for related fragility fractures. Fortunately, WHO has provided new strategies for identifying populations with a high ten-year fracture risk, which together with increasingly sensitive diagnostic methods make it feasible for decision makers in this field to design cost effective fracture prevention strategies. These strategies are aimed at preventing falls and improving bone strength and therefore diminishing the prevalence and incidence of new or recurrent osteoporosis related fractures. Herein we review the content of these new strategies, and the medical treatments available, as well as their efficacy in the Mexican context. Several countries are now reporting a decreasing incidence and prevalence of osteoporosis related fractures, after 30 years of clinical and population-based interventions. Mexico has several effective anti-fracture drug treatments available. Such drugs can be classified according to the mechanism that makes them effective as: 1) antidestructive or anticatabolic, 2) bone forming or anabolic, and 3) those with both actions or mixed drugs. The authors argue that treatment strategies that use drugs to strengthen bone tissue must assure normal mineralization of the already formed, remnant bone tissue and/or the newly formed bone tissue in order to encourage biochemical outcomes like formation of mature hydroxyapatite crystals with complete biomechanical and biochemical properties and therefore long term benefits. The present review includes some perspectives that will surely enhance osteoporosis management in the near future and which will bring about a decrease in the impact of the problems in Mexico. PMID- 19287886 TI - Measurement of bone mineral in vivo: an improved method. 1963. PMID- 19287887 TI - The definition and diagnosis of osteoporosis. 1987. PMID- 19287888 TI - Quantitative ultrasound and bone health. AB - This review of quantitative ultrasound (QUS) and bone health uses the current literature to summarise the clinical and research effectiveness of QUS. QUS has been demonstrated to have the ability to predict fracture, particularly at the hip. However, the magnitude of prediction is fracture-site, measurement-site and device dependent. The correlations between dual X-ray absorptiometry (DXA) and bone mineral density (BMD) are weak to moderate, resulting in different subjects being identified as being at risk of fracture by the two different methods. QUS is sensitive to age and menopause-related changes and to clinical risk factors and lifestyle factors associated with osteoporosis. Whilst a limited ability of QUS to monitor therapeutic intervention has been demonstrated, this is still an area where it's poorer precision, in comparison to DXA, results in limited applicability. Whilst DXA remains the gold standard for the diagnosis of osteoporosis, QUS may be of use for the prediction of those at risk of future fracture in areas where there is limited availability of DXA. PMID- 19287889 TI - Osteoporosis in Mexico: 'the challenge'. PMID- 19287890 TI - Quantitative ultrasound for the detection and management of osteoporosis. AB - Quantitative ultrasound (QUS) appears to be developing into an acceptable, low cost and readily-accessible alternative to dual X-ray absorptiometry (DXA) measurements of bone mineral density (BMD) in the detection and management of osteoporosis. Perhaps the major difficulty with their widespread use is that many different QUS devices exist that differ substantially from each other, in terms of the parameters they measure and the strength of empirical evidence supporting their use. But another problem is that virtually no data exist outside of Caucasian or Asian populations. In general, heel QUS appears to be most tested and most effective. Some, but not all heel QUS devices are effective assessing fracture risk in some, but not all populations, the evidence being strongest for Caucasian females > 55 years old, though some evidence exists for Asian females > 55 and for Caucasian and Asian males > 70. Certain devices may allow to estimate the likelihood of osteoporosis, but very limited evidence exists supporting QUS use during the initiation or monitoring of osteoporosis treatment. Likely, QUS is most effective when combined with an assessment of clinical risk factors (CRF); with DXA reserved for individuals who are not identified as either high or low risk using QUS and CRF. However, monitoring and maintenance of test and instrument accuracy, precision and reproducibility are essential if QUS devices are to be used in clinical practice; and further scientific research in non Caucasian, non-Asian populations clearly is compulsory to validate this tool for more widespread use. PMID- 19287891 TI - The impact of methods for estimating bone health and the global burden of bone disease. AB - Osteoporosis constitutes a major public health problem through its association with age related fractures. Fracture rates are generally higher in caucasian women than in other populations. Important determinants include estrogen deficiency in women, low body mass index, cigarette smoking, alcohol consumption, poor dietary calcium intake, physical inactivity, certain drugs and illnesses. Thus, modification of physical activity and dietary calcium/vitamin D nutrition should complement high risk approaches. In addition, the recently developed WHO algorithm for evaluation of 10-year absolute risk of fracture provides a means whereby various therapies can be targeted cost-effectively to those at risk. Risk factors, together with bone mineral density (BMD) and biochemical indices of bone turnover, can be utilised to derive absolute risks of fracture and cost-utility thresholds at which treatment is justified. These data will provide the basis for translation into coherent public health strategies aiming to prevent osteoporosis both in individuals and in the general population. PMID- 19287893 TI - The clinical and epidemiologic consequences of redefining treatment criteria: who should be treated? AB - Bone mineral density (BMD) is the tool for diagnosing osteoporosis in older adults. However, BMD alone is not sufficient for deciding who should be given treatment at either the individual patient or the public health level. Robust, scientifically validated algorithms that combine BMD with other clinical risk factors provide more accurate assessment of fracture probability. New guidelines for managing osteoporosis are now based on the assessment of absolute fracture risk, not simply on bone mineral density values. Accordingly, treatment resources will be redirected away from young postmenopausal women with low BMD and low fracture risk toward older adults at moderate or high risk for fracture. It is expected that, with these algorithms, the cost and effectiveness of medical care for patients with osteoporosis will be improved. PMID- 19287894 TI - The importance and relevance of peak bone mass in the prevalence of osteoporosis. AB - Bone mass and strength achieved at the end of the growth period, simply designated as 'Peak Bone Mass (PBM)', plays an essential role in the risk of osteoporotic fractures occurring in adulthood. It is considered that an increase of PBM by one standard deviation would reduce the fracture risk by 50%. As estimated from twin studies, genetics is the major determinant of PBM, accounting for about 60 to 80% of its variance. During pubertal maturation, the size of the bone increases whereas the volumetric bone mineral density remains constant in both genders. At the end of puberty, the sex difference is essentially due to a greater bone size in male than female subjects. This is achieved by larger periosteal deposition in boys, thus conferring at PBM a better resistance to mechanical forces in men than in women. Sex hormones and the IGF-1 system are implicated in the bone sexual dimorphism occurring during pubertal maturation. The genetically determined trajectory of bone mass development can be modulated to a certain extent by modifiable environmental factors, particularly physical activity, calcium and protein intakes. Prepuberty appears to be an opportune time to modify environmental factors that impinge on bone mineral mass acquisition. PMID- 19287895 TI - Epidemiology of osteoporosis in Latin America 2008. AB - Osteoporosis is a growing disease in Latin America, rates of fractures actually in LA are quite similar to the ones in Southern Europe and slightly lower than USA and Northern Europe. According to the population projection of the WHO subjects over 65 of age will increase significantly in the next 50 years in LA and the number of hip fractures in this area will equal the numbers in Europe and USA. Unfortunately, in LA there are not adequate resources in terms of medical facilities for osteoporosis diagnosis, treatment or rehabilitation. Moreover, there are no policies of reimbursement for osteoporosis densitometry, bone markers measurements or medications in many countries or it is very limited in the ones who have started some kind of specific program for this matters. Thus, we have to face this public health problem by increasing the awareness of the disease in the general population, specific risk groups, schools, journalists or politicians. Improve medical education at different levels: Medical students, General Practitioners, Specialists, Nurses, and Social Workers. And, we have to develop enough influence to Public and Private Health Systems in order to make them to develop specific osteoporosis programs to cover a wide range of the population in LA, who actually is absolutely abandon of this kind of support. PMID- 19287896 TI - Reference values for areal bone mineral density among a healthy Mexican population. AB - OBJECTIVE: Compare the influence of ethnicity in the prevalence of osteopenia and osteoporosis in various Mexican populations using two normal dual X-ray absorptiometry (DXA) reference databases: manufacturer's incorporating US Hispanic population and a normal mestizo Mexican population. MATERIAL AND METHODS: MMP included 9 946 subjects participating in an ongoing long-term cohort study focusing on lifestyle and chronic diseases, of which 6 487 MMP males and females aged 7 to 80 years were the normal subjects used to determine bone density T- and Z-scores, following WHO criteria, and peak bone mass values. Abnormal bone mass density values estimated by the manufacturer's and peak bone mass reference values were compared. RESULTS AND CONCLUSIONS: Our results show that by using the manufacturer's T-score values in the mestizo Mexican population we are underestimating the number of abnormal bone mass BMD populations. PMID- 19287897 TI - Peak bone mass and bone mineral density correlates for 9 to 24 year-old Mexican women, using corrected BMD. AB - OBJECTIVE: To determine the age of peak bone mass (PBM) in Mexican women and factors associated with both BMDa and corrected BMD (BMDcorr) at the femoral neck and the spine (L2-L4). MATERIAL AND METHODS: Data on 461 women between 9 and 24 years old was used. An interview was performed and height and weight were measured. BMDa was measured by a densitometer and BMDcorr by the method proposed by Kroger et al. (1992). RESULTS: PBM at the spine (L2-L4) was observed later than at the femoral neck. Both BMDa and BMDcorr at the lumbar spine correlate with age, socio-economic status, body fat percentage and height. BMDa at the femoral neck correlates with overweight and obesity, body fat percentage, height and moderate physical activity; the same variables were associated with BMDcorr except for height. CONCLUSIONS: The method proposed by Kroger et al. was more precise at the femoral neck than at the spine. PMID- 19287898 TI - Correlation trends for bone mineral density in Mexican women: evidence of familiar predisposition. AB - OBJECTIVE: Genetic factors determine bone mineral density (BMD) and peak bone density between 20 and 30 years of age, as well as bone mineral loss after menopause. BMD is a predictor of fractures due to osteoporosis and the impact of genetic factors on osteoporosis. The variation in BMD for each individual is determined by an underlying genetic structure, common genetic effects, particularly with respect to compact bones as compared to those that are primarily trabecular. This article presents the correlation of BMD by anatomical site among different samples of Mexican grandmothers, mothers and granddaughters of mixed race. MATERIAL AND METHODS: The present analysis was performed of healthy employees and their healthy relatives from three different health and academic institutions: the Instituto Mexicano del Seguro Social and the Instituto Nacional de Salud Publica, both located in Cuernavaca, Morelos, as well as the Universidad Autonoma del Estado de Mexico. We selected family-related female participants in order to obtain pairs of mothers and daughters and, whenever possible, grandmother-mother-daughter groups. We were able to match 591 mother daughter pairs for analysis. Additionally, we were able to include grandmothers to create grandmother-mother-daughter triads for further analysis. Bone density measurements were performed of the non-dominant proximal femur, the lumbar spine (L1-L4) and the whole body using a dual X-ray absorptiometry (DXA) Lunar DPX NT instrument. RESULTS: This study included 591 granddaughters, 591 mothers and 69 grandmothers; mean ages were 20, 47 and 72 years old, respectively. A close relationship existed with respect to body mass index (BMI) between mothers and grandmothers (27.9 vs. 27.3). The largest proportion of body fat mass was observed in the group of mothers (28.5%), but was also high in grandmothers (25.7%) and granddaughters (21.1%). The percentage of lean body mass was similar among the three family groups. The correlation of BMD between mothers and grandmothers was greatest for subtotal BMD (0.44) and was very high for the hips (0.39). Using predictive models for hip BMD among grandmothers, mothers and grandchildren, we observed that hip BMD of grandmothers is a predictor of BMD in mothers, with a beta of 0.46 (p 0.001, CI95% 0.19-0.73); (R(2): 0.41). A predictor of BMD of the lumbar spine in grandchildren is BMD of the lumbar spine in mothers (beta 0.30 CI95% 0.07-0.53). CONCLUSIONS: The results obtained in this study suggest that daughters whose mothers have a low BMD for their age will tend to develop the same condition. This indicates the importance of monitoring for girls and adolescent females whose mothers have problems related to osteopenia or osteoporosis. It will therefore be necessary to conduct studies to identify the most significant genes and specific anatomical sites among our population for the purpose of establishing the polymorphic variants for high-risk in the Mexican population. PMID- 19287899 TI - Immunodominance: a new hypothesis to explain parasite escape and host/parasite equilibrium leading to the chronic phase of Chagas' disease? AB - Intense immune responses are observed during human or experimental infection with the digenetic protozoan parasite Trypanosoma cruzi. The reasons why such immune responses are unable to completely eliminate the parasites are unknown. The survival of the parasite leads to a parasite-host equilibrium found during the chronic phase of chagasic infection in most individuals. Parasite persistence is recognized as the most likely cause of the chagasic chronic pathologies. Therefore, a key question in Chagas' disease is to understand how this equilibrium is established and maintained for a long period. Understanding the basis for this equilibrium may lead to new approaches to interventions that could help millions of individuals at risk for infection or who are already infected with T. cruzi. Here, we propose that the phenomenon of immunodominance may be significant in terms of regulating the host-parasite equilibrium observed in Chagas' disease. T. cruzi infection restricts the repertoire of specific T cells generating, in some cases, an intense immunodominant phenotype and in others causing a dramatic interference in the response to distinct epitopes. This immune response is sufficiently strong to maintain the host alive during the acute phase carrying them to the chronic phase where transmission usually occurs. At the same time, immunodominance interferes with the development of a higher and broader immune response that could be able to completely eliminate the parasite. Based on this, we discuss how we can interfere with or take advantage of immunodominance in order to provide an immunotherapeutic alternative for chagasic individuals. PMID- 19287900 TI - The corporate bias and the molding of prescription practices: the case of hypertension. AB - Drug management of hypertension has been a noticeable example of the influence of the pharmaceutical industry on prescription practices. The worldwide leading brands of blood pressure-lowering agents are angiotensin receptor-blocking agents, although they are considered to be simply substitutes of angiotensin converting enzyme (ACE) inhibitors. Commercial strategies have been based on the results of clinical trials sponsored by drug companies. Most of them presented distortions in their planning, presentation or interpretation that favored the drugs from the sponsor, i.e., corporate bias. Atenolol, an ineffective blood pressure agent in elderly individuals, was the comparator drug in several trials. In a re-analysis of the INSIGHT trial, deaths appeared to have been counted twice. The LIFE trial appears in the title of more than 120 reproductions of the main and flawed trial, as a massive strategy of scientific marketing. Most guidelines have incorporated the corporate bias from the original studies, and the evidence from better designed studies, such as the ALLHAT trial, have been largely ignored. In trials published recently corporate influences have touched on ethical limits. In the ADVANCE trial, elderly patients with type 2 diabetes and cardiovascular disease or risk factors, allocated to placebo, were not allowed to use diuretic and full doses of an ACE inhibitor, despite the sound evidence of benefit demonstrated in previous trials. As a consequence, they had a 14% higher mortality rate than the participants allocated to the active treatment arm. This reality should be modified immediately, and a greater independence of the academy from the pharmaceutical industry is necessary. PMID- 19287901 TI - Proximal tubular dysfunction as an indicator of chronic graft dysfunction. AB - New strategies are being devised to limit the impact of renal sclerosis on graft function. Individualization of immunosuppression, specifically the interruption of calcineurin-inhibitors has been tried in order to promote better graft survival once chronic graft dysfunction has been established. However, the long term impact of these approaches is still not totally clear. Nevertheless, patients at higher risk for tubular atrophy and interstitial fibrosis (TA/IF) development should be carefully monitored for tubular function as well as glomerular performance. Since tubular-interstitial impairment is an early event in TA/IF pathogenesis and associated with graft function, it seems reasonable that strategies directed at assessing tubular structural integrity and function would yield important functional and prognostic data. The measurement of small proteins in urine such as alpha-1-microglobulin, N-acetyl-beta-D-glucosaminidase, alpha/pi S-glutathione transferases, beta-2 microglobulin, and retinol binding protein is associated with proximal tubular cell dysfunction. Therefore, its straightforward assessment could provide a powerful tool in patient monitoring and ongoing clinical assessment of graft function, ultimately helping to facilitate longer patient and graft survival associated with good graft function. PMID- 19287902 TI - FANCD2 Western blot as a diagnostic tool for Brazilian patients with Fanconi anemia. AB - Fanconi anemia is a rare hereditary disease showing genetic heterogeneity due to a variety of mutations in genes involved in DNA repair pathways, which may lead to different clinical manifestations. Phenotypic variability makes diagnosis difficult based only on clinical manifestations, therefore laboratory tests are necessary. New advances in molecular pathogenesis of this disease led researchers to develop a diagnostic test based on Western blot for FANCD2. The objective of the present study was to determine the efficacy of this method for the diagnosis of 84 Brazilian patients with Fanconi anemia, all of whom tested positive for the diepoxybutane test, and 98 healthy controls. The FANCD2 monoubiquitinated isoform (FANCDS+/FANCD2L-) was not detected in 77 patients (91.7%). In 2 patients (2.4%), there was an absence of both the monoubiquitinated and the non-ubiquitinated proteins (FANCD2S-/FANCD2L-) and 5 patients (5.9%) had both isoforms (FANCD2S+/FANCD2L+). This last phenotype suggests downstream subtypes or mosaicism. All controls were diepoxybutane negative and were also negative on the FANCD2 Western blot. The Western blot for FANCD2 presented a sensitivity of 94% (79/84) and specificity of 100% (98/98). This method was confirmed as an efficient approach to screen Brazilian patients with deleterious mutations on FANCD2 (FANCD2S-/FANCD2L-) or other upstream genes of the FA/BRCA pathway (FANCDS+/FANCD2L-), to confirm the chromosome breakage test and to classify patients according to the level of FA/BRCA pathway defects. However, patients showing both FANCD2 isoforms (FANCD2S+/FANCD2L+) require additional studies to confirm mutations on downstream Fanconi anemia genes or the presence of mosaicism. PMID- 19287903 TI - A retrospective comparison of cyclophosphamide plus antithymocyte globulin with cyclophosphamide plus busulfan as the conditioning regimen for severe aplastic anemia. AB - Allogeneic hematopoietic stem cell transplantation (AHSCT) is the treatment of choice for young patients with severe aplastic anemia (SAA). The association of antithymocyte globulin (ATG) and cyclophosphamide (CY) is the most frequently used conditioning regimen for this disease. We performed this retrospective study in order to compare the outcomes of HLA-matched sibling donor AHSCT in 41 patients with SAA receiving cyclophosphamide plus ATG (ATG-CY, N = 17) or cyclophosphamide plus busulfan (BU-CY, N = 24). The substitution of BU for ATG was motivated by the high cost of ATG. There were no differences in the clinical features between the two groups, including age, gender, cytomegalovirus status, ABO match, interval between diagnosis and transplant, and number of total nucleated cells infused. No differences were observed in the time to neutrophil and platelet engraftment, or in the risk of veno-occlusive disease and hemorrhage. However, there was a higher risk of mucositis in the BU-CY group (71 vs 24%, P = 0.004). There were no differences in the incidence of neutrophil and platelet engraftment, acute and chronic graft-versus-host disease, and transplant related mortality. There was a higher incidence of late rejection in the ATG-CY group (41 vs 4%, P = 0.009). Although the ATG-CY group had a longer follow-up (101 months) than the BU-CY group (67 months, P = 0.04), overall survival was similar between the groups (69 vs 58%, respectively, P = 0.32). We conclude that the association BU-CY is a feasible option to the conventional ATG-CY regimen in this population. PMID- 19287904 TI - Mechanical cardiac remodeling and new-onset atrial fibrillation in long-term follow-up of subjects with chronic Chagas' disease. AB - Atrial fibrillation (AF) affects subjects with Chagas' disease and is an indicator of poor prognosis. We investigated clinical, echocardiographic and electrocardiographic variables of Chagas' disease in a long-term longitudinal study as predictors of a new-onset AF episode lasting >24 h, nonfatal embolic stroke and cardiac death. Fifty adult outpatients (34 to 74 years old, 62% females) staged according to the Los Andes classification were enrolled. During a follow-up of (mean +/- SD) 84.2 +/- 39.0 months, 9 subjects developed AF (incidence: 3.3 +/- 1.0%/year), 5 had nonfatal stroke (incidence: 1.3 +/- 1.0%/year), and nine died (mortality rate: 2.3 +/- 0.8%/year). The progression rate of left ventricular mass and left ventricular ejection fraction was significantly greater in subjects who experienced AF (16.4 +/- 20.0 g/year and 8.6 +/- 7.6%/year, respectively) than in those who did not (8.2 +/- 8.4 g/year; P = 0.03, and -3.0 +/- 2.5%/year; P = 0.04, respectively). In univariate analysis, left atrial diameter >/=3.2 cm (P = 0.002), pulmonary arterial hypertension (P = 0.035), frequent premature supraventricular and ventricular contraction counts/24 h (P = 0.005 and P = 0.007, respectively), ventricular couplets/24 h (P = 0.002), and ventricular tachycardia (P = 0.004) were long-term predictors of AF. P-wave signal-averaged ECG revealed a limited long-term predictive value for AF. In chronic Chagas' disease, large left atrial diameter, pulmonary arterial hypertension, frequent supraventricular and ventricular premature beats, and ventricular tachycardia are long-term predictors of AF. The rate of left ventricular mass enlargement and systolic function deterioration impact AF incidence in this population. PMID- 19287905 TI - Effect of three exercise programs on patients with chronic obstructive pulmonary disease. AB - We compared the effect of three different exercise programs on patients with chronic obstructive pulmonary disease including strength training at 50_80% of one-repetition maximum (1-RM) (ST; N = 11), low-intensity general training (LGT; N = 13), or combined training groups (CT; N = 11). Body composition, muscle strength, treadmill endurance test (TEnd), 6-min walk test (6MWT), Saint George's Respiratory Questionnaire (SGRQ), and baseline dyspnea (BDI) were assessed prior to and after the training programs (12 weeks). The training modalities showed similar improvements (P > 0.05) in SGRQ-total (ST = 13 +/- 14%; CT = 12 +/- 14%; LGT = 11 +/- 10%), BDI (ST = 1.8 +/- 4; CT = 1.8 +/- 3; LGT = 1 +/- 2), 6MWT (ST = 43 +/- 51 m; CT = 48 +/- 50 m; LGT = 31 +/- 75 m), and TEnd (ST = 11 +/- 20 min; CT = 11 +/- 11 min; LGT = 7 +/- 5 min). In the ST and CT groups, an additional improvement in 1-RM values was shown (P < 0.05) compared to the LGT group (ST = 10 +/- 6 to 57 +/- 36 kg; CT = 6 +/- 2 to 38 +/- 16 kg; LGT = 1 +/- 2 to 16 +/- 12 kg). The addition of strength training to our current training program increased muscle strength; however, it produced no additional improvement in walking endurance, dyspnea or quality of life. A simple combined training program provides benefits without increasing the duration of the training sessions. PMID- 19287906 TI - Exercise may cause myocardial ischemia at the anaerobic threshold in cardiac rehabilitation programs. AB - Myocardial ischemia may occur during an exercise session in cardiac rehabilitation programs. However, it has not been established whether it is elicited when exercise prescription is based on heart rate corresponding to the anaerobic threshold as measured by cardiopulmonary exercise testing. Our objective was to determine the incidence of myocardial ischemia in cardiac rehabilitation programs according to myocardial perfusion SPECT in exercise programs based on the anaerobic threshold. Thirty-nine patients (35 men and 4 women) diagnosed with coronary artery disease by coronary angiography and stress technetium-99m-sestamibi gated SPECT associated with a baseline cardiopulmonary exercise test were assessed. Ages ranged from 45 to 75 years. A second cardiopulmonary exercise test determined training intensity at the anaerobic threshold. Repeat gated-SPECT was obtained after a third cardiopulmonary exercise test at the prescribed workload and heart rate. Myocardial perfusion images were analyzed using a score system of 6.4 at rest, 13.9 at peak stress, and 10.7 during the prescribed exercise (P < 0.05). The presence of myocardial ischemia during exercise was defined as a difference > or = 2 between the summed stress score and summed rest score. Accordingly, 25 (64%) patients were classified as ischemic and 14 (36%) as nonischemic. MIBI-SPECT showed myocardial ischemia during exercise within the anaerobic threshold. The 64% prevalence of ischemia observed in the study should not be looked on as representative of the whole population of patients undergoing exercise programs. Changes in patient care and exercise programs were implemented as a result of our finding of ischemia during the prescribed exercise. PMID- 19287907 TI - Demographic and metabolic characteristics of individuals with progressive glucose tolerance. AB - We evaluated changes in glucose tolerance of 17 progressors and 62 non progressors for 9 years to improve our understanding of the pathogenesis of type 2 diabetes mellitus. Changes in anthropometric measurements and responses to an oral glucose tolerance test (OGTT) were analyzed. We identified 14 pairs of individuals, one from each group, who were initially normal glucose tolerant and were matched for gender, age, weight, and girth. We compared initial plasma glucose and insulin curves (from OGTT), insulin secretion (first and second phases) and insulin sensitivity indices (from hyperglycemic clamp assay) for both groups. In the normal glucose tolerant phase, progressors presented: 1) a higher OGTT blood glucose response with hyperglycemia in the second hour and a similar insulin response vs non-progressors; 2) a reduced first-phase insulin secretion (2.0 +/- 0.3 vs 2.3 +/- 0.3 pmol/L; P < 0.02) with a similar insulin sensitivity index and a lower disposition index (3.9 +/- 0.2 vs 4.1 +/- 0.2 micromol.kg-1.min 1 ; P < 0.05) vs non-progressors. After 9 years, both groups presented similar increases in weight and fasting blood glucose levels and progressors had an increased glycemic response at 120 min (P < 0.05) and reduced early insulin response to OGTT (progressors, 1st: 2.10 +/- 0.34 vs 2nd: 1.87 +/- 0.25 pmol/mmol; non-progressors, 1st: 2.15 +/- 0.28 vs 2nd: 2.03 +/- 0.39 pmol/mmol; P < 0.05). Theses data suggest that beta-cell dysfunction might be a risk factor for type 2 diabetes mellitus. PMID- 19287908 TI - Factors associated with bacteremia due to multidrug-resistant Gram-negative bacilli in hematopoietic stem cell transplant recipients. AB - The epidemiology of bacteremia developing during neutropenia has changed in the past decade, with the re-emergence of Gram-negative (GN) bacteria and the development of multidrug resistance (MDR) among GN bacteria. We conducted a case control study in order to identify factors associated with bacteremia due to multidrug-resistant Gram-negative (MDRGN) isolates in hematopoietic stem cell transplant recipients. Ten patients with MDRGN bacteremia were compared with 44 patients with GN bacteremia without MDR. Bacteremia due to Burkholderia or Stenotrophomonas sp was excluded from analysis (3 cases), because the possibility of intrinsical resistance. Infection due to MDRGN bacteria occurred in 2.9% of 342 hematopoietic stem cell transplant recipients. Klebsiella pneumoniae was the most frequent MDRGN (4 isolates), followed by Pseudomonas aeruginosa (3 isolates). Among non-MDRGN, P. aeruginosa was the most frequent agent (34%), followed by Escherichia coli (30%). The development of GN bacteremia during the empirical treatment of febrile neutropenia (breakthrough bacteremia) was associated with MDR (P < 0.001, odds ratio = 32, 95% confidence interval = 5_190) by multivariate analysis. Bacteremia due to MDRGN bacteria was associated with a higher death rate by univariate analysis (40 vs 9%; P = 0.03). We were unable to identify risk factors on admission or at the time of the first fever, but the occurrence of breakthrough bacteremia was strongly associated with MDRGN bacteria. An immediate change in the antibiotic regimen in such circumstances may improve the prognosis of these patients. PMID- 19287909 TI - Hospital strain colonization by Staphylococcus epidermidis. AB - The skin and mucous membranes of healthy subjects are colonized by strains of Staphylococcus epidermidis showing a high diversity of genomic DNA polymorphisms. Prolonged hospitalization and the use of invasive procedures promote changes in the microbiota with subsequent colonization by hospital strains. We report here a patient with prolonged hospitalization due to chronic pancreatitis who was treated with multiple antibiotics, invasive procedures and abdominal surgery. We studied the dynamics of skin colonization by S. epidermidis leading to the development of catheter-related infections and compared the genotypic profile of clinical and microbiota strains by pulsed field gel electrophoresis. During hospitalization, the normal S. epidermidis skin microbiota exhibiting a polymorphic genomic DNA profile was replaced with a hospital-acquired biofilm producer S. epidermidis strain that subsequently caused repetitive catheter related infections. PMID- 19287910 TI - Effects on prolactin secretion and binding to dopaminergic receptors in sleep deprived lupus-prone mice. AB - Sleep disturbances have far-reaching effects on the neuroendocrine and immune systems and may be linked to disease manifestation. Sleep deprivation can accelerate the onset of lupus in NZB/NZWF(1) mice, an animal model of severe systemic lupus erythematosus. High prolactin (PRL) concentrations are involved in the pathogenesis of systemic lupus erythematosus in human beings, as well as in NZB/NZWF(1) mice. We hypothesized that PRL could be involved in the earlier onset of the disease in sleep-deprived NZB/NZWF(1) mice. We also investigated its binding to dopaminergic receptors, since PRL secretion is mainly controlled by dopamine. Female NZB/NZWF(1) mice aged 9 weeks were deprived of sleep using the multiple platform method. Blood samples were taken for the determination of PRL concentrations and quantitative receptor autoradiography was used to map binding of the tritiated dopaminergic receptor ligands [3H]-SCH23390, [3H]-raclopride and [3H]-WIN35,428 to D(1) and D(2) dopaminergic receptors and dopamine transporter sites throughout the brain, respectively. Sleep deprivation induced a significant decrease in plasma PRL secretion (2.58 +/- 0.95 ng/mL) compared with the control group (25.25 +/- 9.18 ng/mL). The binding to D(1) and D(2) binding sites was not significantly affected by sleep deprivation; however, dopamine transporter binding was significantly increased in subdivisions of the caudate-putamen- posterior (16.52 +/- 0.5 vs 14.44 +/- 0.6), dorsolateral (18.84 +/- 0.7 vs 15.97 +/- 0.7) and ventrolateral (24.99 +/- 0.5 vs 22.54 +/- 0.7 microCi/g), in the sleep-deprived mice when compared to the control group. These results suggest that PRL is not the main mechanism involved in the earlier onset of the disease observed in sleep-deprived NZB/NZWF(1) mice and the reduction of PRL concentrations after sleep deprivation may be mediated by modifications in the dopamine transporter sites of the caudate-putamen. PMID- 19287911 TI - Interfaces for noninvasive ventilation: does it matter? PMID- 19287912 TI - Semiquantitative echocardiographic evaluation of intrapulmonary vascular dilatations: correlation with evaluation of shunt levels and pulmonary function parameters. AB - OBJECTIVE: To correlate semiquantitative evaluation of intrapulmonary vascular dilatations (IPVD) with quantitative evaluation of shunt levels, as well as to describe clinical and pulmonary function findings in a sample of liver disease patients with IPVD. METHODS: Patients presenting transthoracic echocardiography (TTE) positivity for IPVD underwent clinical evaluation, pulmonary function tests and pulmonary shunt quantification (scintigraphy with technetium-99m-labeled albumin macroaggregates and blood gas analysis after pure oxygen breathing). RESULTS: A total of 28 liver cirrhosis patients were studied (mean age, 47.5 years; 60.7% were Child-Pugh class B). A 4-point, ascending scale was used as a measure of IPVD intensity, which was scored as 1, 2, 3 and 4, respectively, in 13 (46.4%), 9 (32.1%), 2 (7.1%) and 4 (14.3%) of the patients. Patients were divided into a low-intensity group (scores 1 and 2) and a high-intensity group (scores 3 and 4). The mean shunt assessed using scintigraphy was 14.9% in the sample as a whole and was lower in the low-intensity group (11.7% vs. 26.3%; p = 0.01). The mean shunt by blood gas analysis was higher in the high-intensity group (8.3% vs. 16.3%; p < 0.001). Mean PaO2 was lower in the high-intensity group. There was a negative correlation between DLCO and IPVD severity (r = -0.406, p = 0.01). CONCLUSIONS: TTE is a safe, useful tool for assessing IPVD severity in liver disease patients. The IPVD intensity assessed using TTE correlated with the intrapulmonary shunt values obtained through the quantitative methods evaluated, as well as with pulmonary gas exchange abnormalities. PMID- 19287913 TI - Risk factors and prophylaxis for venous thromboembolism in hospitals in the city of Manaus, Brazil. AB - OBJECTIVE: To identify and classify risk factors for venous thromboembolism (VTE) in hospitalized patients, as well as to evaluate medical practices regarding prophylaxis for the disease. METHODS: An observational cross-sectional study, carried out between January and March of 2006, involving inpatients at three hospitals in the city of Manaus, Brazil. Risk stratification for VTE was based on the criteria established by the Brazilian Society of Angiology and Vascular Surgery and by the International Union of Angiology. Clinical, surgical and medication-related risk factors were analyzed. The statistical analysis of the data obtained was conducted, adopting an alpha error of 5% and 95% CI. Qualitative data were analyzed using the chi-square test, whereas quantitative data were analyzed using Student's t-test. RESULTS: Of the 1,036 patients included (total number of admissions, 1,051), 515 (49.7%) were male, and 521 (50.3%) were female. A total of 23 risk factors for VTE were identified (total number of occurrences, 2,319). The stratified risk for VTE was 50.6%, 16.6% and 30.8% among the admissions of high-, moderate- and low-risk cases, respectively. In 73.3% of the admissions, nonpharmacological prophylaxis was not employed at any point during the study period. In 74% of those classified as high- or moderate-risk cases, no prophylactic medications were administered. CONCLUSIONS: This study showed that, in the population studied, risk factors were common and that prophylactic measures were not employed in patients prone to developing VTE and its complications. PMID- 19287914 TI - Expectations and patient satisfaction related to the use of thoracotomy and video assisted thoracoscopic surgery for treating recurrence of spontaneous primary pneumothorax. AB - OBJECTIVE: To compare the outcomes of thoracotomy and video-assisted thoracoscopic surgery (VATS) in the treatment of recurrence of primary spontaneous pneumothorax. METHODS: Medical records of patients presenting recurrence of primary spontaneous pneumothorax were retrospectively reviewed. Patients were divided into two groups: those who underwent conservative thoracotomy (n = 53, thoracotomy group); and those who underwent VATS (n = 47, VATS group). RESULTS: Although there were no deaths in either group and the length of hospital stays was similar between the two, there was greater morbidity in the thoracotomy group. Patients in the thoracotomy group required more pain medication for longer periods than did those in the VATS group (p < 0.05). In the thoracotomy group, the rate of recurrence was 3%. Pain was classified as insignificant at one month after the operation by 68% of patients in the VATS group and by only 21% of those in the thoracotomy group (p < 0.05). At three years after the surgical procedure, 97% of the VATS group patients considered themselves completely recovered from the operation, compared with only 79% in the thoracotomy group (p < 0.05). Chronic or intermittent pain, requiring the use of analgesics more than once a month, was experienced by 90% of the thoracotomy group patients and 3% of the VATS group patients. In addition, 13% of the thoracotomy group patients required clinical pain management. CONCLUSIONS: We recommend VATS as the first-line surgical treatment for patients with recurrent primary spontaneous pneumothorax. PMID- 19287915 TI - Long-term repercussions of a pulmonary rehabilitation program on the indices of anxiety, depression, quality of life and physical performance in patients with COPD. AB - OBJECTIVE: To assess the 24-month effects of a pulmonary rehabilitation program (PRP) on anxiety, depression, quality of life and physical performance of COPD patients. METHODS: Thirty patients with COPD (mean age, 60.8 +/- 10 years; 70% males) participated in a 12-week PRP, which included 24 physical exercise sessions, 24 respiratory rehabilitation sessions, 12 psychotherapy sessions and 3 educational sessions. All patients were evaluated at baseline (pre-PRP), at the end of the treatment (post-PRP) and two years later (current) by means of four instruments: the Beck Anxiety Inventory; the Beck Depression Inventory; Saint George's Respiratory Questionnaire; and the six-minute walk test (6MWT). RESULTS: The comparison between the pre-PRP and post-PRP values revealed a significant decrease in the levels of anxiety (pre-PRP: 10.7 +/- 6.3; post-PRP: 5.5 +/- 4.4; p = 0.0005) and depression (pre-PRP: 11.7 +/- 6.8; post-PRP: 6.0 +/- 5.8; p = 0.001), as well as significant improvements in the distance covered on the 6MWT (pre-PRP: 428.6 +/- 75.0 m; post-PRP: 474.9 +/- 86.3 m; p = 0.03) and the quality of life index (pre-PRP: 51.0 +/- 15.9; post-PRP: 34.7 +/- 15.1; p = 0.0001). There were no statistically significant differences between the post-PRP and current evaluation values. CONCLUSIONS: The benefits provided by the PRP in terms of the indices of anxiety, depression and quality of life, as well as the improved 6MWT performance, persisted throughout the 24-month study period. PMID- 19287916 TI - Nutritional, clinical and socioeconomic profile of patients with cystic fibrosis treated at a referral center in northeastern Brazil. AB - OBJECTIVE: To describe the profile of patients with cystic fibrosis (CF). METHODS: A prospective, cross-sectional study involving CF patients < 18 years of age, evaluated between March and July of 2006 at a referral center in northeastern Brazil. Nutritional assessment was performed using Z scores for height/age (H/A), weight/age (W/A) and weight/height (W/H), as well as %weight/height (%W/H) and body composition measurements. Socioeconomic and clinical data were obtained. RESULTS: Twenty-one patients were evaluated, 12 (57.1%) of whom were female. Mean age at diagnosis was 3.8 +/- 3.9 years. The principal features at diagnosis were respiratory infection (85.7%), steatorrhea (66.7%) and nutritional deficit (47.6%). The mean Z scores for W/A, H/A and W/H were 0.73 +/- 0.28, 0.34 +/- 0.21 and 0.73 +/- 0.35, respectively. Mean %W/H was 94.52 +/- 1.58. The percentage of malnourished children assessed by Z score differed from that assessed by %W/H (nutritional deficit in 66.7% and 33.3%, respectively; p > 0.05). Socioeconomic status, clinical status and Shwachman score were better among well-nourished patients than among those classified as malnourished (p < 0.05 for Shwachman score). CONCLUSIONS: Normal nutritional status was identified based on nutritional indicators (W/A, H/A and W/H), whereas nutritional deficit was identified by assessing body composition. Socioeconomic factors proved favorable, especially maternal education and per capita income. Age at diagnosis was higher than that reported in the literature, although the Shwachman score and the incidence of respiratory infections demonstrated that the patients presented good clinical status. PMID- 19287917 TI - Effectiveness of a photogrammetric model for the analysis of thoracoabdominal respiratory mechanics in the assessment of isovolume maneuvers in children. AB - OBJECTIVE: To test the applicability of a geometric model, adapted to the supine position, for the analysis of respiratory mechanics regarding changes in lateral thoracoabdominal areas in children with asthma. METHODS: Nineteen children (mean age, 11.26 +/- 1.28 years) performed isovolume maneuvers (IVMs) after maximal inspiration, followed by glottal closure and alternation of airflow between the abdominal and thoracic compartments. The maneuvers were recorded in a digital video camera placed perpendicularly to the movement plane, and the images of interest were selected. The geometric model was traced on each image based on surface landmarks of anatomical references. The traced areas were calculated using a computer program, and the results were converted into metric units (cm(2)) using a surface landmark of a known area. Relative contributions (RCs) of the subcompartments in relation to their original compartments and to the chest wall (CW) were calculated. RESULTS: The model was based on 55 thoracic IVM images and 55 abdominal IVM images. Areas and subareas were compared between the maneuvers. There were significant differences in all subcompartments (p < 0.001). All of the RCs were significantly different for the CW (p < 0.001) but not for the ratios between the subcompartments and their original compartments. CONCLUSIONS: This geometric model, applied in children and adapted to the supine position, was effective in profiling changes in the thoracoabdominal silhouette during the IVMs, and the selected subdivisions were useful for the identification of areas contributing the most and the least to CW composition. PMID- 19287918 TI - Comparison between PEF values obtained from a population sample in the city of Sao Carlos, Brazil, and reference values. AB - OBJECTIVE: To compare the reference values for PEF suggested by other authors in 1963, 1989 and 2001 (for populations in the USA, England and Cuba, respectively) with those obtained from a population sample in the city of Sao Carlos, Brazil, and to determine whether there is concordance among them. METHODS: A total of 243 volunteers (123 females and 120 males; 20-70 years of age) participated in the study. The PEF measurements were performed with the volunteer standing, using a nose clip, by means of a portable peak flow meter. These measurements were compared with the reference values using the Friedman test and Dunn's post-hoc test (p < 0.05). RESULTS: Significant differences were found in all age groups from both genders regarding the values predicted in 1989; the same occurred in the 20-30 and 31-40 age groups (both genders), as well as in the 61-70 age group (females only), regarding those predicted in 2001, as well as in the 20-30 age group (males only) regarding those predicted in 1963. CONCLUSIONS: The values predicted in 1963 are appropriate for a population of individuals with the same characteristics as the study sample, except for males in the 20-30 age group. Our study is relevant due to the fact that our sample was larger than that evaluated in the 1963 study. The majority of the values predicted in 1989 and 2001 overestimated the PFE values obtained in our study, proving to be inappropriate for the population studied. PMID- 19287919 TI - Characteristics of patients with lung cancer in the city of Manaus, Brazil. AB - OBJECTIVE: To analyze the characteristics of patients with lung cancer. METHODS: A retrospective descriptive study of patients receiving a histopathological diagnosis of lung cancer between 1995 and 2002 in the city of Manaus, Brazil. Data were collected from the medical archives of three hospitals. Statistical analyses were carried out, and survival curves were generated by means of an actuarial estimator. RESULTS: Of the 352 patients selected, 262 (74.4%) were male and 90 (25.6%) were female. The mean age was 62 years. The following histological types were identified: squamous cell carcinoma, 62.8%; adenocarcinoma, 24.7%; small cell carcinoma, 9.1%; and large cell carcinoma, 3.4%. The most common stages were stages IIIB and IV, in 45% and 21.5%, respectively. Of the total sample, 73.4% were submitted to treatment. Of these, 51.4% underwent radiotherapy; 16.6%, surgery; 15.8%, chemotherapy; and 16.2%, radiotherapy in association with chemotherapy. Cumulative survival rates were low: three-year survival was 6.5%, and five-year survival was 3.5%. CONCLUSIONS: In this group of patients with lung cancer, survival rates were considerably lower than those reported in the literature. This might be attributable to the limited access to the specialized health care system and the advanced stage of the disease at diagnosis. PMID- 19287920 TI - Influence of total face, facial and nasal masks on short-term adverse effects during noninvasive ventilation. AB - OBJECTIVE: Failure of noninvasive ventilation (NIV) has been associated with short-term adverse effects related to the use of masks. The aim of this study was to compare the incidence, type and intensity of adverse effects, as well as the comfort, of total face masks (TFMs), facial masks (FMs) and nasal masks (NMs) during NIV. METHODS: This was a randomized crossover trial involving 24 healthy volunteers submitted to six sessions of NIV in bilevel positive airway pressure mode using the TFM, FM and NM masks at low and moderate-to-high pressure levels. A written questionnaire was applied in order to evaluate eleven specific adverse effects related to the use of the masks. Comfort was assessed using a visual analog scale. The CO2 exhaled into the ventilator circuit was measured between the mask and the exhalation port. RESULTS: The performance of the TFM was similar to that of the NM and FM in terms of comfort scores. Higher pressure levels reduced comfort and increased adverse effects, regardless of the mask type. When the TFM was used, there were fewer air leaks and less pain at the nose bridge, although there was greater oronasal dryness and claustrophobia. Air leaks were most pronounced when the FM was used. The partial pressure of exhaled CO2 entering the ventilator circuit was zero for the TFM. CONCLUSIONS: The short-term adverse effects caused by NIV interfaces are related to mask type and pressure settings. The TFM is a reliable alternative to the NM and FM. Rebreathing of CO2 from the circuit is less likely to occur when a TFM is used. PMID- 19287921 TI - Incidence of TB diagnosed in the emergency room of a teaching hospital in southeastern Brazil. AB - In this study, we analyzed the number of TB cases in an emergency room (ER) and the susceptibility profile of Mycobacterium tuberculosis strains. Patients were selected from among those treated at the Hospital das Clinicas, in the city of Belo Horizonte, Brazil. Between 2002 and 2005, 240 TB patients were identified. Of those, 117 patients (48.7%) were diagnosed in the ER, 72 (61.5%) presenting positive sputum smear microscopy. Drug susceptibility testing was carried out in 90 strains, of which 80 (89%) were sensitive, 9 (10%) were resistant, and 1 (1%) was multidrug resistant. The incidence of positive smear sputum microscopy and resistant TB strains was high, which calls for the immediate adoption of TB control measures in the ER. PMID- 19287922 TI - Pulmonary arterial hypertension and thyroid disease. AB - Recent studies have suggested an association between pulmonary arterial hypertension (PAH) and thyroid diseases (hypothyroidism and hyperthyroidism). This combination has a good prognosis, because the increase in the pulmonary artery pressure is usually slight and reverses after the treatment of the thyroid disease. Although the exact mechanism involved in the pathogenesis of this combination has not yet been established, it has been hypothesized that thyroid hormones and autoimmunity have a direct influence. Due to the high prevalence of thyroid disease in patients with PAH, thyroid function tests should be considered in the investigation of every patient with PAH. In this review, we describe the prevalence of PAH in patients with thyroid diseases and the prevalence of thyroid disease in patients with PAH, as well as addressing the principal effects that thyroid diseases have on the respiratory system. In addition, we report the treatment effects in patients with these diseases. PMID- 19287923 TI - A rare case of synchronous malignant thoracic tumors. AB - Malignant neurogenic mediastinal tumors in adults are uncommon and extremely aggressive. We report the case of a 61-year-old male patient with the simultaneous occurrence of malignant mediastinal schwannoma and bronchioloalveolar carcinoma. Although bronchioloalveolar carcinoma is present in 4-7% of the resected synchronous thoracic tumors, this association has never been reported in the literature. However, it is a common finding in patients presenting apparently inflammatory infiltrates and ground-glass opacities, as in the case presented here. PMID- 19287924 TI - Small cell carcinoma in Pancoast syndrome. AB - Pancoast syndrome consists of signs and symptoms resulting from a tumor affecting the pulmonary apex and adjacent structures. The process is typically caused by a neoplasm. The majority of cases of Pancoast syndrome are caused by bronchogenic carcinoma. The most commonly found histologic subtypes are adenocarcinoma and epidermoid carcinoma. There have been very few reports of small cell lung carcinoma in the genesis of Pancoast syndrome. We describe the case of a patient with Pancoast syndrome caused by small cell lung carcinoma and discuss the aspects related to the diagnosis and treatment. PMID- 19287925 TI - Simultaneous bilateral spontaneous pneumothorax in an adult patient with cystic fibrosis. AB - Pneumothorax is a common complication in cystic fibrosis and is associated with worsening of lung function. However, bilateral simultaneous pneumothorax in cystic fibrosis is a rare condition. We describe the case of a 17-year-old female with cystic fibrosis who presented with spontaneous pneumothorax. Clinically, she presented right-sided chest pain and progressive breathlessness. The pneumothorax failed to resolve after the initial treatment (chest drainage). However, the patient was later successfully treated with additional chest drainage and talc pleurodesis. We also discuss the etiology and management of pneumothorax in patients with cystic fibrosis, since pneumothorax is associated with increased morbidity and mortality among such patients. PMID- 19287926 TI - Epidemiological profile of hepatitis C in blood donors at the Uberaba Regional Blood Center. AB - The aim of the present study was to outline the serological profile of hepatitis C among blood donors seen at the Uberaba Regional Blood Center, Hemominas Foundation, over the last 14 years. The frequency of hepatitis C was compared between first-time and repeat donors and the epidemiological characteristics of those with positive and indeterminate ELISA anti-HCV (third and fourth generation) were analyzed based on the donor histories kept in the archives of the Uberaba Regional Blood Center. The serological ineligibility rate was 0.3%, with higher prevalence in the group of first-time donors. We did not find any significant differences regarding age, skin color, marital status or place of residence between eligible and ineligible donors; however, the frequency of positive serology was higher among men. The lower (0.3%) rate of ineligibility due to hepatitis C that was observed at the Uberaba Regional Blood Center, in relation to most Brazilian blood centers, is probably due to the large number of repeat donors (83.3%). This reinforces the importance of achieving donor commitment for increasing transfusion safety. PMID- 19287927 TI - Pulmonary changes during acute experimental murine mansoni schistosomiasis. AB - Dry cough, dyspnea and manifestations of bronchial asthma have recently been observed in patients with acute schistosomiasis. To investigate the type and pathogenesis of these conditions, an experimental mouse model for acute schistosomiasis was used. Forty mice were divided into four groups of ten each: three infected groups and a non-infected control group. The animals were examined 7, 28-35 and 40 days after exposure to cercariae. During the acute phase of the infection (28-35 days), a process of multifocal interstitial pneumonitis involving the peribronchial, peribronchiolar and subpleural tissues was found. This process was not seen during the other phases of the infection. Indirect immunofluorescence failed to demonstrate the presence of schistosomal antigens in the acute-phase lesions. The pneumonitis was attributed to products (inflammatory mediators) from acute-phase periovular necrotic-inflammatory lesions in the liver that were transported to the lungs by the bloodstream. PMID- 19287928 TI - [Epidemiological profile of rabies in the northwestern region of Sao Paulo State, from 1993 to 2007]. AB - The epidemiology of animal rabies in the region of Aracatuba, in the northwest of Sao Paulo State, from 1993 to 2007, is described according to the results from diagnoses made at laboratories in the region, using the fluorescent antibody and mouse inoculation tests. Out of 10,579 samples analyzed, 4.9% were positive (518/10,579). Dogs accounted for 67% of the cases (346/518) and these occurred between 1993 and 1997. Among the other positive samples, 16% (84/518) were in cattle and 9.7% (50/518) were in bats. Among the 42 municipalities in the region, 23 (55%) presented at least one rabies-positive case, while 13 of them had cases in bats. Three distinct cycles of rabies were identified in the northwestern region of the State of Sao Paulo: the urban cycle characterized predominantly by canine rabies (1993 to 1997); and the aerial and rural cycles starting in 1998, with predominance of cases in bats in urban areas and in herbivores. PMID- 19287929 TI - [Antigen profile of rabies virus isolated from different species of non hematophagous bats in the region of Presidente Prudente, State of Sao Paulo]. AB - Using the monoclonal antibody technique, the Clinical and Molecular Virology Laboratory of the Institute of Biomedical Sciences of the University of Sao Paulo typed 18 rabies virus samples from non-hematophagous bats of several species from the region of Presidente Prudente, SP, Brazil. Among these samples, 15 (82.3%) were defined as variant 3 (compatible with samples isolated from Desmodus rotundus bats) and three (16.7%) as variant 4 (compatible with samples isolated from Tadarida brasiliensis bats). PMID- 19287930 TI - [Dengue in children: clinical and epidemiological characteristics, Manaus, State of Amazonas, 2006 and 2007]. AB - This study presents the clinical and epidemiological aspects of children affected with dengue, in Manaus, AM, in 2006 and 2007. There were 482 The aim of this study was to present the clinical and epidemiological characteristics of children affected by dengue, in Manaus, State of Amazonas, in 2006 and 2007. Analysis was conducted on 482 dengue cases with confirmation from laboratory tests, in 2006 (46.9% in children under 15 years of age), and 1,538 cases in 2007 (57.7% in children under 15 years of age). The data were obtained from the National System for Notifiable Diseases (Sistema Nacional de Agravos de Notificacao; SINAN) in Manaus. The variables analyzed were: age, gender, district where subject lived, month when symptoms started, serotypes and clinical form of the disease. Statistically significant increases in the proportion of cases in children under 15 years of age and in the proportion of severe forms were found between 2006 and 2007. Although the hemorrhagic form was almost three times more frequent in 2007, the mortality rate was lower. The serotypes identified were DEN-3 in 2006 and DEN 1, DEN-2 and DEN-3 in 2007. In 2007, cases were recorded in every month of the year. PMID- 19287931 TI - [Vaccination against yellow fever among patients on immunosuppressors with diagnoses of rheumatic diseases]. AB - Yellow fever is endemic in some countries. The anti-yellow fever vaccine is the only effective means of protection but is contraindicated for immunocompromised patients. The aim of this paper was to report on a case series of rheumatological patients who were using immunosuppressors and were vaccinated against this disease. This was a retrospective study by means of a questionnaire applied to these patients, who were vaccinated 60 days before the investigation. Seventy patients of mean age 46 years were evaluated. Most of them were female (90%). There were cases of rheumatoid arthritis (54), systemic lupus erythematosus (11), spondyloarthropathy (5) and systemic sclerosis (2). The therapeutic schemes included methotrexate (42), corticosteroids (22), sulfasalazine (26), leflunomide (18), cyclophosphamide (3) and immunobiological agents (9). Sixteen patients (22.5%) reported some minor adverse effect. Among the eight patients using immunobiological agents, only one presented a mild adverse effect. Among these patients using immunosuppressors, adverse reactions were no more frequent than among immunocompetent individuals. This is the first study on this topic. PMID- 19287932 TI - [Prevalence of human T-cell lymphotropic virus (HTLV-1/2) infection among puerperae in Cuiaba, Mato Grosso, 2006]. AB - The prevalence of human T-cell lymphotropic virus (HTLV-1/2) infection among puerperae in the State of Mato Grosso, Brazil, is unknown. Through this cross sectional study, the prevalence of HTLV-1/2 infection among puerperae attended at three public maternity hospitals in Cuiaba, State of Mato Grosso, was defined. Between April and September 2006, 3,831 deliveries took place and 2,965 puerperae underwent serological tests for HTLV-1/2: enzyme-linked immunosorbent assay (ELISA) and western blot. The mean age of the women studied was 23.9 years. The prevalence of HTLV-1/2 was 0.2%, i.e. similar to the prevalence observed in the general population of many developed centers in Brazil. This finding of low prevalence suggests that there is still no justification for introducing public health interventions for the population of pregnant women in our setting, to reduce the vertical transmission of HTLV-1/2. PMID- 19287933 TI - [Changes to the prevalence of cervical squamous intraepithelial lesions and risk factors among HIV/AIDS patients attended at a specialized service in southern Brazil from 1995 to 1999 and 2006 to 2007]. AB - The aim of this work was to study changes to the prevalence of cervical squamous intraepithelial lesions (CSIL) among HIV1-positive women following the introduction of highly active antiretroviral therapy (HAART), and to evaluate changes to the prevalence of risk factors for CSIL. Fifty patients were studied in 1995-1999 and 120 patients in 2006-2007. Demographic, behavioral and laboratorial data were collected. The prevalences of CSIL and other risk factors were calculated for the two periods. The prevalence of CSIL was 66% in the first period and 43% in the second period (p = 0.007). The mean CD4 in 1995-1999 was 275.71 (SD 283.23) and in 2006-2007 it was 463.32 (SD 231.90; p=0.001). There were significant changes to the factors of age, skin color, marital status and smoking between the two periods. The decrease in the prevalence of CSIL may be related to the use of the HAART strategy and to changes to the risk factors for CSIL over time. PMID- 19287934 TI - [Assessment of an entomological surveillance system for Chagas disease with community participation in Mambai and Buritinopolis, State of Goias]. AB - Entomological surveillance of Chagas disease in Mambai and Buritinopolis, in the State of Goias, Brazil, has been kept up through the local population's participation, consisting of reporting the presence of vectors inside their homes. A long time has elapsed since instituting these control measures and it has now been certified that vector transmission has been halted. Thus, this study sought to evaluate the population's knowledge and practices in this situation. The results show that there has been progressive indifference towards the topic of 'Chagas disease', which can be attributed to the reduction in the magnitude of the problem that this disease represented, little participation in surveillance among schools, low epidemiological importance of secondary and native vectors and, consequently, limited control interventions from health services in response to notifications. It is proposed that direct search activities by means of sampling should be carried out periodically, and that there should be greater involvement among teaching institutions. PMID- 19287935 TI - [Socioenvironmental factors associated with the spatial distribution of malaria in the Vale do Amanhecer settlement, Municipality of Juruena, State of Mato Grosso, 2005]. AB - Malaria is a disease with focal distribution. In Brazil, settlement and gold mining areas in the Legal Amazon region present a great concentration of cases. This paper analyzes the spatial distribution of malaria cases taking into consideration environmental and social factors in the Vale do Amanhecer settlement in the municipality of Juruena, Mato Grosso, Brazil. In 2005, 359 autochthonous cases were notified in the settlement. Using the Kernel method, areas of greater and lesser intensities of case numbers were identified. The areas of greater intensity presented 290 cases and those of lesser intensity, 64 cases. The intensity of distribution varied within the settlement, thus indicating areas of great intensity of cases favoring transmission, like gold mining areas. Therefore, although the settlement was considered to be the main focus for malaria, there were specific factors within it that, once identified, could contribute towards the disease control. PMID- 19287936 TI - [Mortality trend due to accidental tetanus from 1981 to 2004 in Pernambuco and analysis of the impact on intensive care unit attendance]. AB - Despite reductions in the incidence of accidental tetanus cases in Brazil, there has not been any significant decrease in its mortality. In this case series, the mortality rates before and after establishing standard management practices for tetanus patients in the intensive care unit at the Oswaldo Cruz University Hospital are compared over the period from 1981 to 2004. Over these 24 years, 1.971 patients were admitted. Before establishing the intensive care unit management, the mortality rate was 35%. The Intensive care unit for attending to tetanus patients was established in 1997. From 1998 to 2004, the mortality rate fell to 12.6%: OR = 0.27 (95% CI = 0.18-0.39); p < 0.001. This trend was seen in all age groups and both sexes. The centralization of attendance for these patients into a single specialized service with early treatment in an intensive care unit has therefore been decisive in reducing the mortality rate. This service can count on the medical team's vast experience of tetanus management, with better treatment of symptoms that forestalls the serious complications from this disease. PMID- 19287937 TI - [Study on virulence factors associated with biofilm formation and phylogenetic groupings in Escherichia coli strains isolated from patients with cystitis]. AB - Escherichia coli samples isolated from female patients with cystitis were characterized with regard to the presence of virulence factors associated with biofilm formation and phylogenetic groupings. Polymerase chain reaction results demonstrated that all the samples were positive for the gene fimH (type 1 fimbriae), 91 for fliC (flagellins), 50 for papC (P fimbriae), 44 for kpsMTII (capsules) and 36 for flu (antigen 43). The results from assays to quantify the biofilm formation demonstrated that 44 samples produced biofilm on polystyrene microplates and 56 samples produced weak or no biofilm. We also confirmed that Escherichia coli samples were present in phylogenetic groups B2 and D. PMID- 19287938 TI - [Viability, taxonomic confirmation and enzymatic detection of Acremonium species preserved under mineral oil in the URM Culture Collection]. AB - Hydrolytic enzymes secreted by fungi play an important role in the pathogenesis of infection. With the aim of evaluating the enzymatic activity, 31 isolates of Acremonium stored in the University of Recife Mycology (URM) Culture Collection were tested. Culture fragments were transferred to glycoside broth for reactivation and further growth in potato dextrose agar medium in order to investigate viability and purity and to confirm the taxonomy through observing the macroscopic and microscopic characteristics. To detect enzymes, milk casein and gelatin were used as substrates for proteinase, starch for amylase and soy lecithin for phospholipase. Among the 31 cultures, 26 (83.9%) remained viable and 24 (92.3%) were confirmed taxonomically. Out of these 24 cultures, 12 (50%) presented proteinase activity, of which two (16.7%) were on milk casein, one (8.3%) on gelatin and nine (75%) on both substrates; 16 (66.7%) degraded starch. None of the cultures presented phospholipase activity. It was concluded that Acremonium species are able to produce enzymes that are involved in the pathogenicity of fungal infections. PMID- 19287939 TI - [The assessment process within science and the nomination of Carlos Chagas for the Nobel prize for Physiology or Medicine]. AB - One of the greatest achievements in the history of medicine was the description of Chagas disease by the physician and scientist Carlos Chagas. A hundred years after the discovery of the disease, speculation still remains regarding the two official nominations of Carlos Chagas for the Nobel Prize, the biggest worldwide scientific award, in 1913 and in 1921. It has been accepted that the reason why the prize was not awarded to this brilliant scientist may have been the strong opposition that he faced in Brazil, from some physicians and researchers of that time. They went as far as questioning the existence of Chagas disease, thereby possibly influencing the decision of the Nobel Committee not to award the prize to him. Analysis of the database of the Nobel prize archives, with the revelation of the names of nominators, nominees and prizewinners spanning the years 1901 1951, brought information not only about what was considered to be a scientific achievement at that time, but also about who the important scientists were and what the relationships between them were. The non-recognition of Carlos Chagas' discoveries by the Nobel Committee appears to be more correctly explained by these factors than by the negative impact of the local opposition. PMID- 19287940 TI - [Puerperal septic shock due to beta-hemolytic Streptococcus and Waterhouse Friderichsen syndrome]. AB - An exceptional case of a 15-year-old puerpera with septic shock caused by Group A beta-hemolytic Streptococcus and Waterhouse-Friderichsen syndrome is reported. The findings were observed at the necropsy. The characteristics of the diagnosis, pathogenesis and evolution of this puerperal infection (sepsis), associated with adrenal hemorrhage and insufficiency are reviewed in this paper. PMID- 19287941 TI - [Canine visceral leishmaniasis in Marica, State of Rio de Janeiro: first report of an autochthonous case]. AB - Visceral leishmaniasis is a zoonosis of public health importance, and dogs represent one of the main problems. This paper describes the first autochthonous case of canine visceral leishmaniasis in the municipality of Marica. It provides new facts regarding the geographical distribution of Leishmania (Leishmania) chagasi in the State of Rio de Janeiro. PMID- 19287942 TI - [Algid malaria: a syndromic diagnosis]. AB - Two patients with malaria due to Plasmodium falciparum who progressed to shock syndrome are reported. They received hemodynamic support in an intensive care unit without using antibiotics and presented improvements. Algid malaria should be a syndromic diagnosis of varying etiology (dehydration, bacterial infection, bleeding and/or adrenal insufficiency). PMID- 19287943 TI - [Impact of low-intensity laser on the suppression of infections caused by Herpes simplex viruses 1 and 2: in vitro study]. AB - The use of low-level laser to suppress infections caused by Herpes simplex viruses 1 and 2 was evaluated after one to five applications. A gradual reduction in replication of Herpes simplex viruses 1 and 2 was observed, with 68.4% and 57.3% inhibition, respectively, after five applications, thus favoring its clinical use. PMID- 19287944 TI - Severe cytomegalic pneumonitis associated with pulmonary aspergillosis in a child with immunosuppression due to chemotherapy for treating pineoblastoma. PMID- 19287945 TI - Molecular principles of cancer invasion and metastasis (review). AB - The main threat and the reason for most cancer deaths are not the primary neoplasias, but secondary tumors, the metastases. Drastic phenotypic and biochemical changes occur during the metamorphosis of a normal tissue cell into an invasive cancer cell. These alterations concern various areas such as growth factor signaling, cell-cell adhesion, gene expression, motility or cell shape. Cancer cells of epithelial origin can even shed their typical qualities and characteristics and adopt a mesenchymal-like phenotype. This is often referred to as an epithelial-mesenchymal transition. Various oncogenes, tumor suppressor genes and metastasis suppressor genes are known to affect the invasiveness and the metastatic potential of tumor cells. Cells of the innate and adaptive immunity, adjacent stroma cells as well as chemokines and their receptors also play a vital role in the spread of cancer cells. Furthermore, the micro environment, vascularization and the supply with special cytokines affect the above-mentioned changes. Finally, some researchers claim that tumors consist of two types of cells - transit amplifying cells and cancer stem cells. Only the latter are thought to be able to proliferate indefinitely and thus they might be the cells that successfully spread and initially build most of the clinically relevant metastases. This review article describes some of the molecular principles which underlie those changes as well as covers some aspects of current research. PMID- 19287946 TI - Uterine cervical carcinoma: role of matrix metalloproteinases (review). AB - Epidemiological and experimental studies have provided evidence that human papillomavirus (HPV) infection is a main player in the development of uterine cervical neoplasms. Migration of cancer cells from the origin tissue to surrounding or distant organs is essential for tumor progression. Many studies of tumor invasion and metastases have focused on the degradation of the extracellular matrix where matrix metalloproteinases (MMPs) play a central role. Two of these enzymes, MMP-2 and MMP-9, have been correlated with the processes of tumor cell invasion and metastasis in human cancers, including uterine neoplasms. It has been shown that the up-regulation of MMPs is associated with progression of cervical uterine neoplasms. This review describes the current understanding of MMP-2 and MMP-9 expression and activity in pre-cancer and cancer lesions of cervical uterine, which may open new strategies for diagnostic and therapeutic interventions. PMID- 19287947 TI - Irinotecan-induced ovarian follicular apoptosis is attenuated by deleting the kinase domain of death-associated protein kinase. AB - Although death-associated protein kinase (DAPK) is a Ca2+/calmodulin-regulated serine/threonine kinase that plays important roles in various types of apoptotic cell death, there have been no reports of its tissue distributions and functions in female reproductive organs. By comparing C57BL/6 wild-type mice with DAPK mutant mice lacking the 74-amino acid catalytic kinase domain of DAPK, the cellular distributions and biological functions of DAPK in murine ovaries were investigated. In situ hybridization analyses with sense and antisense riboprobes revealed that DAPK mRNA was selectively and highly expressed in granulosa cells in the ovaries of both types of mice. There were no significant differences in the body weights, ovarian weights and unstimulated ovarian follicular numbers between the wild-type and DAPK-mutant mice. Intraperitoneal injection of CPT-11, an anticancer topoisomerase I inhibitor that causes granulosa cell-specific apoptosis partly through Fas-Fas ligand (FasL) interactions in MCH mice, induced follicular apoptosis in both the wild-type and DAPK-mutant mice. However, the numbers of apoptotic follicles were significantly reduced in the DAPK-mutant mice. The Fas and FasL expression levels in the CPT-11-injected mice did not differ significantly between the wild-type and DAPK-mutant mice. These results indicate that DAPK positively regulates intracellular signaling pathways for CPT 11-induced granulosa cell apoptosis. PMID- 19287948 TI - High CCR7 mRNA expression of cancer cells is associated with lymph node involvement in patients with esophageal squamous cell carcinoma. AB - Esophageal squamous cell carcinoma (SCC) is one of the biological malignant tumors. Once a tumor invades the submucosa, an incidence of lymph node (LN) metastases is very high, thus resulting in poor survival. Recently, chemokines have been reported to play an important role in organ-specific metastases in several malignancies. In particular, CCR7 has been reported to be associated with LN metastases by immunohistochemistry. However, there have been no studies of quantitative analyses of CCR7 mRNA expression on cancer cells. In this study, we investigated the clinical significance of the expression of CCR7 in the establishment of LN metastases of esophageal SCC. A series of 78 patients with esophageal SCC who underwent esophagectomy were consecutively selected. The expression of CCR7 mRNA from tumor tissue samples was analyzed by quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR), and that from cancer cell samples collected using laser microdissection system was analyzed by qRT-PCR. Immunohistochemical staining of CCR7 was also performed. Although CCR7 mRNA expression in tumor tissues demonstrated no association with the LN metastases, that in cancer cells correlated with LN metastases (p<0.05) due to the fact that not only cancer cells but also infiltrating lymphocytes expressed CCR7 in tumor tissue. Multivariate logistic regression analysis revealed a high CCR7 expression in cancer cells to be an independent predictive factor for LN metastases. These results suggested that CCR7 expression might play an important role in establishing LN metastases in patients with esophageal SCC. PMID- 19287949 TI - Significance of monocyte chemoattractant protein-1 in angiogenesis and survival in colorectal liver metastases. AB - Monocyte chemoattractant protein-1 (MCP-1) has been demonstrated to play a role in tumor progression. The present study examined the MCP-1 expression of colorectal liver metastases and determined whether MCP-1 is related to tumor progression and is a predictive marker for survival after hepatic resection of colorectal liver metastases. Eighty-seven patients with colorectal liver metastases were evaluated by immunohistochemistry of MCP-1, Angiopoietin-2, CD68, and CD34 for determination of microvessel density. Clinicopatholgical data were also examined. In a separate experiment, immunohistochemistry of MCP-1 was performed to investigate the expression of primary colorectal tumor according to the clinical stage. MCP-1 mRNA expression was determined in colorectal cancer cell lines. Forty-nine patients (56%) showed high expression of MCP-1 of colorectal liver metastases. High MCP-1 expression was related to multiple colorectal liver metastases. When the degree of MCP-1 expression increased, microvessel density count significantly increased compared with low MCP-1 expression. The MCP-1 expression correlated with Angiopoietin-2 expression. MCP-1 expression of the primary colorectal cancer increased as the clinical stage advanced. The increased MCP-1 mRNA expression was observed in cancer cell lines which have high metastasis potency. Univariate analysis demonstrated that the timing of metastases, tumor size, number of metastases, and MCP-1 expression were significant prognostic factors. Multivariate analysis demonstrated that MCP-1 expression was a significant prognostic factor in hepatic disease-free survival. The MCP-1 expression in colorectal liver metastases, at least in part, may be associated with angiogenesis and be a predictive marker for hepatic recurrence after hepatic resection for colorectal liver metastases. PMID- 19287950 TI - High expression of ncRAN, a novel non-coding RNA mapped to chromosome 17q25.1, is associated with poor prognosis in neuroblastoma. AB - Neuroblastoma shows complex patterns of genetic aberrations including MYCN amplification, deletion of chromosome 1p or 11q, and gain of chromosome 17q. The 17q gain is frequently observed in high-risk neuroblastomas, however, the candidate genes still remain elusive. In the present study, we integrated the data of comparative genomic hybridization of 236 tumors by BAC array and expression profiling of 136 tumors by using the in-house cDNA microarray carrying 5,340 genes derived from primary neuroblastomas. A novel candidate gene mapped to chromosome 17q25.1 with two splicing variants, Nbla10727 and Nbla12061, was identified. The transcript size appeared to be 2.3 kb by Northern blot, however, the cDNA sequences had no obvious open reading frame. The protein product was undetectable by both in vivo and in vitro translation assays, suggesting that the transcript might not encode any protein product. Therefore, we named it as ncRAN (non-coding RNA expressed in aggressive neuroblastoma). In analysis of 70 patients with sporadic neuroblastoma, the high levels of ncRAN mRNA expression were significantly associated with poor outcome of the patients (p<0.001). The multivariate analysis showed that expression of ncRAN mRNA was an independent prognostic factor among age, stage, origin and MYCN expression. Ectopic expression of ncRAN induced transformation of NIH3T3 cells in soft agar, while knockdown of endogenous ncRAN with RNA interference significantly inhibited cell growth in SH-SY5Y cells. Collectively, our results suggest that ncRAN may be a novel non-coding RNA mapped to the region of 17q gain and act like an oncogene in aggressive neuroblastomas. PMID- 19287951 TI - A novel mechanism whereby BRCA1/1a/1b fine tunes the dynamic complex interplay between SUMO-dependent/independent activities of Ubc9 on E2-induced ERalpha activation/repression and degradation in breast cancer cells. AB - BRCA1 dysfunction is associated with hormone-responsive cancers. We have identified a consensus SUMO modification site in the amino-terminal region of BRCA1/1a/1b proteins and the mutation in this potential SUMO acceptor site (K 109 to R) impaired their ability to bind and repress ligand-dependent ERalpha transcriptional activity in breast cancer cells. Furthermore, we have found SUMO E2-conjugating enzyme Ubc9 to bind BRCA1 proteins. We have mapped BRCA1 [within amino acids (aa) 1-182] as the minimum domain that is sufficient for in vitro binding to Ubc9 as well as for regulating ERalpha activity. BRCA1 Mutant #1 (K109 to R) was impaired in its ability to both bind, as well as modulate Ubc9 mediated SUMO-dependent/independent E2-induced ERalpha transcriptional activity in breast cancer cells. Similarly, BRCA1 cancer-predisposing mutation (61Cys-Gly) abrogated the ability to both bind Ubc9 as well as inhibit ERalpha activity suggesting physiological significance. Addition of BRCA1 but not Mutant #1 to E2-induced ERalpha in the presence of SUMO-1 and Ubc9 resulted in the degradation of ERalpha suggesting BRCA1 to be a putative SUMO-1 and Ubc9-dependent E3 ubiquitin ligase for ERalpha. This is the first report demonstrating the participation of Ubc9 in BRCA1 E3 ubiquitin ligase mediated degradation of ERalpha. These results suggest a novel function for BRCA1 in regulating the dynamic cycles of SUMO and ubiquitin modifications required for ERalpha turn over and deregulation of this molecular switch due to lack of BRCA1 results in ERalpha-negative/positive breast cancers. This study will help in designing novel BRCA1 function-based targeted treatment for breast cancers. PMID- 19287952 TI - Cross-infection of tumor cells by contact with T lymphocytes loaded with Newcastle disease virus. AB - Oncolytic virotherapy, a new type of cancer therapy involving viruses with oncolytic and immunostimulatory potential, is based on tumor selective viral replication, resulting in a specific lysis of tumor cells. Effective tumor targeting of oncolytic viruses remains a major problem because only a fraction of systemically applied viruses can reach the tumor tissue. We describe for the first time in an in vitro co-culture system that T lymphocytes can be loaded with Newcastle disease virus (NDV) in such a way that the virus load will be transferred to the tumor target cells upon contact of the T cells with tumor cells. The effectiveness of this NDV 'hitchhiking' on T cells can be influenced by the amount of virus, the ratio of T cells to tumor cells, the activation status of the T cells and by the virulence of the virus as shown by flow cytometry, quantitative real-time PCR and fluorescence microscopy. In a tumor neutralization assay in vitro, monolayers of human tumor cells could be completely and effectively destroyed by the addition of polyclonally activated human T cells loaded with oncolytic NDV. This process involves the formation of large T cell clusters as revealed by phase-contrast microscopy. Loading of oncolytic NDV onto activated T cells and adoptive transfer into a tumor-bearing host might enhance the efficacy of adoptive T cell therapy of tumors as well as tumor targeting of oncolytic viruses. PMID- 19287953 TI - Progressive increase of genetic alteration in urinary bladder cancer by combined allelotyping analysis and comparative genomic hybridization. AB - Bladder cancer is the ninth most common cancer in the world. Urothelial carcinoma (formerly known as transitional cell carcinoma) comprises the majority of bladder cancers. In order to decipher the genetic alteration leading to the carcinogenesis of urothelial cancer, we performed genome-wide allelotyping analysis using 384 microsatellite markers spanning 22 autosomes together with comparative genomic hybridization (CGH) in 21 urothelial cancer. High frequency of allelic imbalance was observed in chromosome arm 1q (61.9%), 3p (61.9%), 4q (66.67%), 8p (57.14%), 9p (76.2%) and 9q (66.67%). Allelic imbalance with frequency above average was also observed in chromosome arm 2q, 10p, 10q, 11p, 11q, 12q, 13q, 15q, 17p and 19q. The allelic imbalance of each case and fractional allelic loss for each chromosome was associated with higher tumor grade and stage (P<0.05). We have also delineated several minimal deletion regions on chromosome 3p, 4q, 8p, 9p, 9q, 11p, 13q, 16q and 17p. By CGH analysis, common chromosomal alterations included gain of 1p, 1q, 12q, 16p, 17q and 19p as well as loss of 4q and 9p in most of the cases. Our findings may provide valuable information to locate putative oncogenes and tumor suppressor genes in the carcinogenesis of bladder cancer in this locality. PMID- 19287954 TI - Expression of RIG-I, IRF3, IFN-beta and IRF7 determines resistance or susceptibility of cells to infection by Newcastle Disease Virus. AB - Newcastle Disease Virus (NDV) is an avian paramyxovirus with anti-neoplastic and immune-stimulatory properties which has raised considerable interest for cancer therapy. To better understand the molecular nature of the tumor selective replication of NDV, we investigated the cellular responses of murine normal and tumor cells after infection by NDV. To this end, we compared the basal expression of different antiviral proteins as well as the expression induced by the addition of NDV to the cells in vitro and in vivo. Primary macrophages were found to be resistant to NDV infection and exhibited a high basal and induced expression of various antiviral genes. In contrast, macrophage-derived RAW tumor cells were highly susceptible to NDV infection and displayed a low expression of several antiviral genes. Macrophage-derived J774 tumor cells were intermediate with regard to NDV replication and antiviral gene expression. The responsiveness to exogenously added IFN-alpha was found highest in normal macrophages, lowest in the RAW cells, and intermediate in the J774 cells. We also analysed dendritic cells as well as additional normal and tumor cell types. A strong inverse correlation was obeserved between the susceptibility to infection and the basal expression of the antiviral genes RIG-I, IRF3, IRF7 and IFN-beta. A strong expression of these genes can explain the resistance of normal cells to NDV infection and a weak antiviral gene expression the broad susceptibility of tumor cells. PMID- 19287955 TI - Quantitative immunocytochemical profile to predict early outcome of disease in triple-negative breast carcinomas. AB - We aimed in this study at identifying prognostic immunohistochemical molecular signatures indicative of disease outcome, also relevant for development of new specific therapies, in triple-negative (ER, PR, c-erbB2- negative) breast carcinoma subtypes. We evaluated 42 markers in tissue micro-arrays from a series of 924 breast carcinomas including 184 triple-negative tumors using standardized quantitative immunocytochemical assays and correlated the data with patients' outcome (mean follow-up of 79 months). When 27/42 markers including basal-like markers first found to be individually significant for prognosis in a univariate analysis (log-rank test) in 924 tumors, were secondly evaluated in the triple negative tumor subtype (184/924), eleven including maspin, P21, P27, PTEN, caveolin, EGFR, FAK, P38, pMAPK, STAT1 and CD10 were 89.2% predictive of disease outcome in logistic regression. When markers reported in the literature as expressed in basal-like subtype were evaluated in the 924 series, only eight (EGFR, CK14, moesin, caveolin, cMet, ckit, CD44v6, C10) were prognosis predictive in univariate analysis (log-rank test) and in logistic regression were predictive of disease outcome in 66.3% independently of ER, PR and c-erbB2 expression and in 72% in triple-negative tumor subset. The results suggest that the category of 'triple-negative' breast carcinomas does not exactly overlap the basal-like subtype, and that immunoprofiling of triple-negative tumors (not similar to that of basal-like tumors) may be helpful to select patients for more aggressive treatment and provides a basis for development of tailored therapy. PMID- 19287956 TI - Inhibition of calcineurin by cyclosporine A exerts multiple effects on human melanoma cell lines HT168 and WM35. AB - The immunosuppressant cyclosporine A (CsA) is a specific pharmacological inhibitor of calcineurin, the Ca2+-calmodulin activated phospho-Ser/Thr-specific protein phosphatase. Although calcineurin-inhibiting compounds are applied for local treatment of psoriasis or atopic dermatitis in dermatological practice, little is known about the functions of calcineurin in epidermis-derived malignancies. We investigated the effects of CsA on two human melanoma cell lines, the metastasis forming HT168 and WM35 established from an RGP primary lesion. CsA of 2 microM lowered the enzyme activity by 50% and caused elevation in both mRNA and protein expression of calcineurin. Cell proliferation was diminished, as well as the cellular morphology and the actin organization were altered in both cell lines. CsA increased cell death moderately in both cell lines and reduced the metabolic activity of HT168 cells, but not that of WM35 cells. CsA also elevated the expressions of both Bcl-2 and ERK1/2. Fibronectin guided migration of HT168 cells was stimulated under the effect of CsA, while that of WM35 cells was reduced, moreover, HT168 cells switched from the expression of beta3 to beta1 integrin, but WM35 cells continued to express beta3. Based on our results we propose a multiple, partly malignancy-dependent role of calcineurin in these melanoma cell lines. PMID- 19287957 TI - Characterization of the c.190T>C missense mutation in BRCA1 codon 64 (Cys64Arg). AB - In the Milan area (Northern Italy), we identified a family characterized by a high prevalence of ovarian and breast cancer cases (5 out of 6 subjects, over 3 generations), and a predominant prevalence of ovarian lesions (4 out of 5 patients). Analysis of BRCA1 and BRCA2 genes allowed the identification of the missense c.190T>C mutation in codon 64 (Cys64Arg) of BRCA1. The aims of the present investigation were to characterize the functional implications of the c.190T>C mutation at the molecular level, and to search whether additional polymorphisms might be linked to the peculiar phenotypic features observed in the Italian pedigree. Molecular modelling studies suggested that substitution of the cysteine 64 with an arginine likely disrupts the architecture of the BRCA1 RING finger domain, responsible for the interaction with BARD1, essential for the tumor-suppressor activity of the BRCA1-BARD1 complex. By splicing site information analysis, exonic splicing enhancer site characterization, and analysis of transcript fragment length and sequence, we showed that the c.190T>C mutation was able to modulate the splicing of exon 5 in a fashion opposite to the c.190T>G transversion, responsible for the functionally-related Cys64Gly amino acid substitution. Genotyping of BRCA1 and BRCA2 in the Italian family revealed the presence of two significant polymorphisms: the cancer-associated c.2612C>T SNP in BRCA1, and the c.-26G>A SNP in the BRCA2 gene, acting as an ovarian cancer risk modifier in carriers of deleterious BRCA1 mutations. Analysis of these SNPs in a genotypically-unrelated Polish family, characterized by prevalent breast neoplasms in carriers of the c.190T>C mutation, revealed a genetic profile consistent with the hypothetic role of both polymorphisms. PMID- 19287958 TI - Functional characterization of p53 in nasopharyngeal carcinoma by stable shRNA expression. AB - Nasopharyngeal carcinoma (NPC) is a high-incidence malignancy in Southern China and Southeast Asia. Although mutation of p53 tumor-suppressor gene is a rare event in NPC, NPC has a high frequency of overexpressed/accumulated p53 protein, which was reported to be dysfunction or inactivation in most of NPC. We report here a functional characterization of p53 in an undifferentiated NPC cell line CNE2. To elucidate the biological function of p53, we employed the RNA interference (RNAi) approach to knockdown the endogenously expressed p53 in CNE2 cells. Interestingly, suppression of p53 expression in CNE2 cells was associated with significant down-regulation of p21WAF1/CIP1 expression and decreased HDM2 protein level in both steady state and genotoxic stress induced by ionizing radiation (IR). Consistent with these biochemical data were the accelerated cell cycle progression and the increased proliferation rate, suggesting that p53 retained growth inhibitory activity in CNE2 cells. Indeed, down-regulation of p53 in CNE2 enhanced the ability of CNE2 cells to grow anchorage-independently in vitro and to develop tumors in vivo. Together with the radioresistance acquired by CNE2sip53 cells, our data indicate that in contrast to a previous study, p53 in this NPC cell line remains functional, which may have an important therapeutical implication. PMID- 19287959 TI - Role of fibrillar Tenascin-C in metastatic pancreatic cancer. AB - Interaction of cancer cells with stroma cells facilitates tumor progression by rebuilding the existing extracellular matrix (ECM) microenvironment. In the tumor, upregulation of Tenascin-C (Tn-C) expression potentially can alter tumor behavior. However, the molecular mechanisms by which tumor-stroma interactions affect the tumor microenvironment have not been well characterized. In this study, we analyzed the expression of fibrillar Tn-C (fTn-C) in human metastatic pancreatic cancers. After co-culturing two pancreatic cancer cell lines, highly metastatic BxPc3 cells and non-metastatic PaCa2 cells, with stromal fibroblasts (SF), we evaluated the roles of matrix metalloproteinase 2 (MMP-2) activation and SF in promoting Tn-C organization. Next, we evaluated whether fibrillar Tn-C promotes pancreatic cancer cell movement using cell adhesion and migration assays. Finally, we observed the relationship between MMP-2 activation and fTn-C formation in vivo by injecting the BxPc3 and PaCa2 cells into nude mice. We found that fTn-C was increased in metastatic pancreatic cancer. The fTn-C expression correlated with MMP-2 activity. In the in vitro co-culture, fTn-C organization was found only in BxPc3/SF co-cultures, and required the participation of active MMP-2. The fTn-C reduced cell adhesion and promote pancreatic cancer cell migration by decreasing the adhesive interactions between integrin alpha6beta1 and the ECM. The in vivo tumorigenesis analysis showed that the fTn-C formation and active MMP-2 were significantly increased in the BxPc3 tumors, compared to the PaCa2 tumors. These results demonstrate that Tn-C deposition into the ECM requires participation of active MMP-2 and SF. The deposited Tn-C could promote pancreatic cancer progression. PMID- 19287960 TI - Prostate cancer as an environmental disease: an ecological study in the French Caribbean islands, Martinique and Guadeloupe. AB - Using a transdisciplinary methodological approach we have conducted a multifactorial analysis in Martinique and Guadeloupe in order to elucidate the aetiology of prostate cancer. In 2002, world age standardized rates of prostate cancer were 152 new cases per 100,000 person-years in the two islands; one of the highest worldwide rates and much higher than those reported for other Caribbean islands and metropolitan France. Using a linear regression analysis, we found that the growth curves of incidence rates for Martinique and metropolitan France have been significantly diverging since 1983. That these curves are not parallel suggests that although a Caribbean genetic susceptibility factor may be involved in carcinogenesis, this factor cannot per se account for the observed growing incidence. On the basis of mapping analysis of soil pollution, we further showed that water contamination by pesticides originates from banana plantations. Moreover, we have established retrospectively that general population subjects investigated in 1972 in Martinique for the presence of organochlorinated pesticides in their adipose tissue had been contaminated by extremely high levels of DDT, DDE, alpha, beta and gammaHCH, aldrin and dieldrin. Our study leads to the conclusion that the growing incidence of prostate cancer cannot be related either to a modification of ethnographic factors nor to a change in lifestyle and therefore suggests that environmental factors such as the intensive and prolonged exposure to carcinogenic, mutagenic and reproductive toxin pesticides may cause prostate cancer. PMID- 19287961 TI - Involvement of p16 and PTCH in pathogenesis of melanoma and basal cell carcinoma. AB - The involvement of two tumor suppressors p16 and Ptch in pathogenesis of cutaneous melanomas and basal cell carcinomas (BCCs) was studied through expression of Ptch and p16 and genetic alterations in 9p21 region (p16) and in 9q22.3 region (PTCH) of chromosome 9. Immunohistochemical analyses of paraffin embedded tissues with Ptch and p16 antibodies, typing for 9q22-q31 and 9p21 region with polymorphic markers and p16 and Ptch mutation detection was done. Higher expression of p16 and Ptch in melanoma and BCC of the skin was frequently detected in studied cases. However, allelic loss of PTCH region occurs more frequently in BCCs than loss of heterozygosity of p16 region. Both types of tumors, BCCs and melanomas, suggest involvement of Hh-Gli signaling pathway, but using different mechanisms. PMID- 19287962 TI - SNS-032 prevents hypoxia-mediated glioblastoma cell invasion by inhibiting hypoxia inducible factor-1alpha expression. AB - Hypoxia and hypoxia inducible factor-1alpha (HIF-1alpha) play a critical role in glioblastoma (GBM) which is characterized by highly aggressive and widespread cell invasion into adjacent normal brain tissue. The purpose of this study was to investigate the effect of the novel aminothiazole com-pound SNS-032 in glioblastoma cell invasion under hypoxic condition. SNS-032 is a potent and selective inhibitor of cyclin-dependent kinases 2, 7 and 9 and inhibits both cell cycle and transcription. We analyzed the effect of SNS-032 (0.5 microM) on HIF 1alpha expression and its major trans-regulating factors including COX-2, VEGF, MMP-2 and uPAR that are involved in cellular invasion in tumor hypoxia. Our observations demonstrate SNS-032: i) inhibited hypoxia-induced U87MG cell invasion and among all the other inhibitors tested, SNS-032 is the most effective, ii) blocked HIF-1alpha mediated transcription of COX-2, MMP-2, VEGF and uPAR expression in U87MG cells in response to hypoxia, iii) blocked HIF 1alpha expression by a proteasome independent pathway. The effects were similar to those observed with HIF-1alpha siRNA which prevented cellular invasion by blocking HIF-1alpha expression and its downstream effectors. Taken together, our data suggest that SNS-032 prevents hypoxia-mediated U87MG cell invasion by blocking the expression of HIF-1alpha and its trans-regulating factors. Our results present an opportunity in controlling highly invasive tumors such as glioblastoma using this novel class of compounds. PMID- 19287963 TI - Interaction between survivin and aurora-B kinase plays an important role in survivin-mediated up-regulation of human telomerase reverse transcriptase expression. AB - Survivin, a member of the apoptosis inhibitor family, shows increased expression in human cancers of various origins. It has been demonstrated that survivin inhibits apoptosis via caspase inhibition and promotes mitosis via aurora-B kinase activation. We recently reported that survivin enhances the expression of human telomerase reverse transcriptase (hTERT), a major determinant of telomerase activity in colon cancer cells. Survivin up-regulates hTERT expression by promoting the expression of specificity protein-1 (Sp1)- and c-Myc-mediated gene transcription via enhancing the phosphorylation of these transcriptional factors. However, the mechanism by which survivin regulates the phosphorylation of Sp1 and c-Myc is not well defined. In the present study, we hypothesized that survivin promotes the phosphorylation of Sp1 and c-Myc by activating aurora-B kinase. Inhibition of this enzyme by introducing small inhibitory RNA attenuated the phosphorylation of Sp1 and c-Myc and resulted in the abolition of the survivin effect on hTERT expression. In addition, blocking survivin phosphorylation at a threonine residue by inhibiting cyclin-dependent kinase 1 caused the dissociation of aurora-B kinase from survivin and attenuated the up-regulation of hTERT expression by survivin. Taken together, these results suggest that the interaction between survivin and aurora-B kinase may be essential for survivin to increase hTERT expression. PMID- 19287964 TI - Over- and under-expressed microRNAs in human colorectal cancer. AB - MicroRNAs (miRNAs) constitute a class of small (21-23 nucleotides) noncoding RNAs that function as post-transcriptional gene regulators. It is becoming increasingly clear that altered miRNA expression correlates with the pathogenesis of cancers. The purpose of this study was to determine the up-regulated miRNAs in human colorectal cancer. Total RNA was isolated from cancer tissues and corresponding noncancerous tissues from surgically resected colorectal cancers. The expression profiles of miRNAs were determined using a miRNA microarray containing 455 human miRNA probes. The expression status of selected miRNAs in paired clinical samples was then investigated by real-time RT-PCR. Twenty-one miRNAs were identified by miRNA array analysis as overexpressed in colorectal cancer tissues compared to normal epithelial tissues. Among them, the expression of miR-31, miR-183, miR-17-5p, miR-18a, miR-20a and miR-92 were confirmed to be significantly higher in cancer tissues than in normal tissues (P<0.05). In contrast, the expression of miR-143 and miR-145 in cancer tissues were significantly lower than in normal tissues (P<0.05). The miR-18a overexpression group tended to have a poorer clinical prognosis than the low expression group (P=0.07). We identified miRNAs that were overexpressed or under-expressed in colorectal cancers and which may be correlated with colorectal carcinogenesis. PMID- 19287965 TI - Induction of apoptosis in human leukemia U937 cells by anthocyanins through down regulation of Bcl-2 and activation of caspases. AB - Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, that exhibit important anti-oxidant and anti-inflammatory actions as well as chemotherapeutic effects. However, little is known concerning the molecular mechanisms by which these activities are exerted. In this study, we investigated the anthocyanins isolated from Vitis coignetiae Pulliat for their potential anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that these anthocyanins inhibit cell viability and induce apoptotic cell death of U937 cells in a dose-dependent manner, as measured by hemocytometer counts, by alteration in the mitochondrial membrane potential, by increases in sub-G1 populations and by DNA ladder formation. Apoptosis of U937 cells by anthocyanins was associated with modulation of expression of Bcl-2 and IAP family members. Consequently, anthocyanin treatment induced proteolytic activation of caspase-3, -8 and -9, and a concomitant degradation of poly(ADP-ribose) polymerase. However, anthocyanin-induced growth inhibition and apoptosis were significantly attenuated in Bcl-2 overexpressing U937 cells. Furthermore, z-DEVD fmk, a caspase-3 specific inhibitor, blocked apoptosis and increased the survival of anthocyanin-treated U937 cells. Taken together, these results show that Bcl-2 and caspases are key regulators of apoptosis in response to anthocyanins in human leukemia U937 cells. PMID- 19287966 TI - CYP1B1, but not CYP1A1, is downregulated by promoter methylation in colorectal cancers. AB - Cytochrome P450 (CYP) 1A1 (CYP1A1) and CYP1B1, dioxin-inducible CYP1s, are associated with carcinogenesis in extrahepatic tissues. CYP1B1 is featured in carcinogenesis of hormone-responsive tissues, where the CYP1B1 level is considerably high. Although aberrant expression of these enzymes is also observed in cancers that are not related to hormone response, their roles in carcinogenesis are not yet fully understood. We examined DNA methylation status of the CpG islands within the 5'-flanking region of the CYP1B1 and CYP1A1 genes in 7 colorectal cancer cell lines and 40 primary colorectal cancers. By bisulfite modified direct sequencing, CYP1B1 gene methylation was detected in 2 cell lines (SW48 and Caco-2) and 2 (5%) cancers, but not in corresponding normal tissues. Treatment of the cells with 5-aza-2'-deoxycytidine revealed a clear increase in the CYP1B1 mRNA levels in SW48 and Caco-2 cells, while the amount of methylated alleles decreased. Only HT29 cells showed a clear increase in CYP1A1 mRNA, although there were no apparent differences in methylation status among these cell lines. None of these cell lines showed significant change in mRNA levels of aryl hydrocarbon receptor (AhR) and AhR nuclear translocator (ARNT), which are known to directly activate CYP1 transcription. This observation suggested that expression of CYP1B1, but not CYP1A1, was downregulated by promoter methylation rather than decreased expression of AhR/ARNT. In conclusion, CpG methylation of the CYP1B1 promoter region epigenetically regulates CYP1B1 expression during development of some colorectal cancers. Moreover, cancers with aberrant CYP1B1 expression might show altered response to procarcinogen metabolism and chemotherapy. PMID- 19287967 TI - Non-genomic inhibitory signaling of beta-carotene in squamous cell carcinoma of the lungs. AB - Studies have suggested that retinoids prevent lung cancer by interacting with nuclear retinoid receptors. However, clinical trials with beta-carotene increased lung cancer mortality. We recently showed that beta-carotene stimulates the proliferation of small airway-derived adenocarcinoma by increasing cAMP signaling. Here, we have tested the hypothesis that beta-carotene may stimulate squamous cell carcinoma cells via similar mechanisms. We determined the effects of beta-carotene in cell lines from squamous cell carcinomas and large airway epithelia on proliferation by MTT assays in the presence and absence of inhibitors. Signaling via cAMP/PKA was measured by immunoassays and PKA activation assay. Phosphorylated ERK1/2 was determined by Western blotting. beta carotene significantly inhibited proliferation and phosphorylation of ERK1/2 by Galphas-mediated signaling involving adenylyl cyclase, cAMP, PKA and ERK1/2. These findings introduce a non-genomic inhibitory mechanism of beta-carotene and emphasize the need for the development of marker-guided lung cancer prevention. PMID- 19287968 TI - In silico prediction and experimental validation of natural antisense transcripts in two cancer-associated regions of human chromosome 6. AB - Antisense transcription has long been recognized as a mechanism involved in the regulation of gene expression. Therefore, several human diseases associated with abnormal patterns of gene expression might display antisense RNA-mediated pathogenetic mechanisms. Such issue could be particularly relevant for cancer pathogenesis, since deregulated gene expression has long been established as a hallmark of cancer cells. Herein, we report on a bioinformatic search for antisense transcription in two cancer-associated regions of human chromosome 6 (6q21 and 6q27). Natural antisense transcripts (NATs) for several genes in both genomic regions were predicted in silico and subsequently validated by strand specific RT-PCR. Detailed experimental validation by quantitative real-time RT PCR of five putative cancer related sense-antisense transcript pairs revealed a single candidate tumor suppressor gene (RPS6KA2) whose expression levels display marked cancer-related changes that are likely mediated by its antisense RNA in a breast cancer cell line model. PMID- 19287969 TI - Survival of patients diagnosed with either colorectal mucinous or non-mucinous adenocarcinoma: a population-based study in Canada. AB - Previous studies have shown conflicting results on the prognosis of colorectal mucinous adenocarcinoma. This study compared prognostic characteristics of patients diagnosed with mucinous and non-mucinous adenocarcinomas in a Canadian series. Analyses were based on 165 colorectal mucinous and 1215 non-mucinous adenocarcinoma patients who were registered at the Ottawa Regional Cancer Centre from 1994 to 1997, with follow-up extending to December 31, 2001. Differences in survival were examined using the relative survival analysis and the Cox proportional hazards model. For colon, rectum and both combined, the distribution for age at diagnosis, stage and treatment of patients with mucinous adenocarcinoma was similar to that of non-mucinous patients (all p > or = 0.12). Patients with mucinous histology had fewer well- or moderately-differentiated tumours than non-mucinous patients (all p < 0.01). Overall, no statistically significant differences were noted in 5-year relative survival between mucinous and non-mucinous carcinoma for colon, rectum and their combination (p > or = 0.35 for each). However, when the stages were considered separately, patients with stage III mucinous carcinoma had worse survival than patients with non-mucinous carcinoma for both sites. Multivariate analysis of combined data for colon and rectal cancers indicated that independent significant prognostic factors were stage for mucinous, with age and grade as well as stage for non-mucinous carcinoma. In conclusion, no significant differences in stage distribution and overall survival were found between mucinous and non-mucinous patients for colorectal cancer. PMID- 19287970 TI - In vitro anti-cancer activity of two ethno-pharmacological healing plants from Guatemala Pluchea odorata and Phlebodium decumanum. AB - Many traditional healing plants successfully passed several hundred years of empirical testing against specific diseases and thereby demonstrating that they are well tolerated in humans. Although quite a few ethno-pharmacological plants are applied against a variety of conditions there are still numerous plants that have not been cross-tested in diseases apart from the traditional applications. Herein we demonstrate the anti-neoplastic potential of two healing plants used by the Maya of the Guatemala/Belize area against severe inflammatory conditions such as neuritis, rheumatism, arthritis, coughs, bruises and tumours. Phlebodium decumanum and Pluchea odorata were collected, dried and freeze dried, and extracted with five solvents of increasing polarity. We tested HL-60 and MCF-7 cells, the inhibition of proliferation and the induction of cell death were investigated as hallmark endpoints to measure the efficiency of anti-cancer drugs. Western blot and FACS analyses elucidated the underlying mechanisms. While extracts of P. decumanum showed only moderate anti-cancer activity and were therefore not further analysed, particularly the dichloromethane extract of P. odorata inhibited the cell cycle in G2-M which correlated with the activation of checkpoint kinase 2, and down-regulation of Cdc25A and cyclin D1 as well as inactivation of Erk1/2. In HL-60 and MCF-7 cells this extract was a very strong inducer of cell death activating caspase-3 followed by PARP signature type cleavage. The initiating death trigger was likely the stabilization of microtubules monitored by the rapid acetylation of alpha-tubulin, which was even more pronounced than that triggered by taxol. The dichloromethane extract of P. odorata contains apolar constituents which inhibit inflammatory responses and exhibit anti-cancer activity. The strong proapoptotic potential warrants further bioassay-guided fractionation to discover and test the active principle(s). PMID- 19287971 TI - Sulforaphane down-regulates COX-2 expression by activating p38 and inhibiting NF kappaB-DNA-binding activity in human bladder T24 cells. AB - Cyclooxygenase-2 (COX-2) overexpression has been associated with the grade, prognosis and recurrence of transitional cell carcinoma (TCC) of the bladder. In this study, sulforaphane, a dietary isothiocyanate, down-regulated COX-2 expression in human bladder transitional cancer T24 cells at both transcriptional and translational levels. Sulforaphane (5-20 microM) induced nuclear translocation of NF-kappaB and reduced its binding to the COX-2 promoter, a key mechanism for suppressing COX-2 expression by sulforaphane. Moreover, sulforaphane increased expression of p38 and phosphorylated-p38 protein. A specific inhibitor of p38 MAPK, SB202190, was used to further investigate its pivotal role in sulforaphane-mediated down-regulation of COX-2. Exposure of T24 cells to SB202190 1 hour prior to sulforaphane treatment abolished the effect of sulforaphane on COX-2 mRNA down-regulation, but enhanced COX-2 transcription. Furthermore, SB202190 alone induced NF-kappaB translocation to the nucleus, promoted NF-kappaB binding to the COX-2 promoter and resulted in up-regulation of COX-2 expression. Taken together, these data suggest that p38 is essential in sulforaphane-mediated COX-2 suppression and provide new insights into the molecular mechanisms of sulforaphane in the chemoprevention of bladder cancer. PMID- 19287972 TI - A natural antisense transcript, BOKAS, regulates the pro-apoptotic activity of human Bok. AB - Bok is a proapoptotic member in the Bcl-2 family and the expression of Bok is regulated by cellular stresses. In the present study, we have isolated and characterized a natural antisense transcript of Bok, BOKAS. The BOKAS gene consists of 2 exons, that expresses as a non-protein coding transcript. Both BOKAS and Bok are transcribed from the same locus but in opposite orientations. The mRNA expression of BOKAS was only detected in testis and certain cancer tissues but not in other normal adult tissues examined. Overexpression of BOKAS alone exhibited no significant anti- or pro-apoptotic activity but it was able to inhibit Bok-induced apoptosis in HeLa cells. Our results suggest that BOKAS may function specifically in the human testis, where it serves as an antisense molecule to regulate Bok-induced apoptosis. In addition, natural antisense transcripts were identified for BAD and BCL2L12. PMID- 19287973 TI - Combination therapy using three novel prolactin receptor antagonist-based fusion proteins effectively inhibits tumor recurrence and metastasis in HER2/neu transgenic mice. AB - Previously, prolactin receptor antagonist (G129R)- based fusion proteins were developed including G129R fusions with an angiogenesis inhibitor (endostatin), an immune system modulator (interleukin 2), and a modified truncated cytotoxin (PE38KDEL). Each fusion protein was designed to target the PRLR-positive cells via the G129R moiety and at the same time attack a hallmark common to cancer cells via the second moiety. In this study, we tested the efficacy of the three fusion proteins as a combination therapy in an aggressive but clinically relevant mouse tumor model. To test the feasibility and to optimize a treatment regimen, allografts of a mammary carcinoma cell line (McNeuA) derived from an MMTV-neu transgenic mouse were first used. Growth of the allografts was significantly retarded by regimens which combined all three fusion proteins. In addition, a significant increase in cytotoxic CD8+ T cells was observed within the tumors of the combination treated groups. After establishing the dosing regimen, two doses of cocktail treatment (low and high doses administered twice weekly) along with individual component controls were administered to female MMTV-neu transgenic mice after surgical removal of a naturally occurring tumor. The average tumor recurrence time was significantly delayed in both low and high combination treatment groups in comparison to the no treatment control group (34, 50 and 18 days, respectively). The total number of lung metastases was also significantly decreased in both combination treatment groups. In conclusion, using G129R-based fusion proteins to target mammary carcinomas and to tackle multiple hallmarks of cancer at the same time was an effective strategy for treating HER2-postive mammary cancer in this mouse tumor model. PMID- 19287974 TI - Messenger RNA expression of COX-2 and angiogenic factors in primary colorectal cancer and corresponding liver metastasis. AB - Several new drugs that are targeted towards various angiogenic factors have shown considerable potential for controlling tumor proliferation and metastases. Expression levels of the targeted genes in primary tumors and metastases should be understood to maximize the use of such drugs. The present study aimed to clarify associations between mRNA levels of cyclooxygenase 2 (COX-2) and angiogenic factors [vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8)] in primary colorectal cancer and in corresponding liver metastasis. We also compared these gene expressions of primary colorectal cancer between patients with and without liver metastasis. In 31 pairs of formalin-fixed and paraffin-embedded primary and metastatic liver tumors as well as 27 specimens of consecutive stage II patients without recurrence, mRNA was quantified by real time reverse transcription-polymerase chain reaction following the laser capture microdissection. We found a significantly positive correlation in IL-8 between primary tumors and matched liver metastases (p=0.034, rs=0.39) and in VEGF (p=0.0083, rs=0.48), but not in COX-2, which was associated with both VEGF (p=0.044, rs=0.37) and IL-8 (p=0.0004, rs=0.64) in primary colorectal cancers. Multiple regression analysis revealed that COX-2 was independently associated with IL-8 (p<0.0001). There were no differences in mRNA levels between patients with and without liver metastasis. The mRNA levels of VEGF and IL-8 in liver metastasis can be predicted from those in primary colorectal cancer. COX-2 might exert angiogenic activity more through the IL-8, than the VEGF pathway. These angiogenic factors were sufficiently up-regulated before hematogenous metastasis. These preliminary data merit further validation studies. PMID- 19287975 TI - Lapatinib, a dual inhibitor of ErbB-1/-2 receptors, enhances effects of combination chemotherapy in bladder cancer cells. AB - Survival rate of patients diagnosed with the invasive form of bladder cancer is low suggesting an urgent need to implement novel treatments. GTC (gemcitabine, paclitaxel and cisplatin) is a new chemotherapeutic regimen, which has shown promise in clinical trials. Given that receptor tyrosine kinases of the ErbB family are overexpressed in a high proportion of metastatic bladder tumours, approaches involving small-molecule inhibitors of ErbB receptors in combination with conventional cytostatic drugs are of potential interest. Here, we show that the dual inhibitor of ErbB receptors, lapatinib, enhances cytostatic and induces cytotoxic effects of GTC in two bladder cancer cell lines which differ with regard to expression levels of proteins taking part in the ErbB pathway. Lapatinib inhibited phosphorylation of ErbB receptors and also reduced the level of phosphorylated AKT. Flow cytometry analysis demonstrated that GTC treatment affects cell cycle distribution differently in the presence or absence of lapatinib. In RT112 cells, which express high levels of ErbB receptors and harbour wild-type p53, combined GTC/lapatinib treatment resulted in the phosphorylation of p53 at Ser46 and accumulation of sub-G1 cell populations. Our data indicate that a combinatorial approach involving GTC and lapatinib may have therapeutic potential in a subset of bladder tumours depending on the genetic context. PMID- 19287976 TI - Defective expression of deoxycytidine kinase in cytarabine-resistant acute myeloid leukemia cells. AB - Resistance to cytarabine (Ara-C) incapacitates the therapeutic effort during the treatment of acute myeloid leukemia (AML). To elucidate mechanism responsible for the development of resistance to Ara-C, we established the Ara-C resistant AML 2/WT cell sublines, AML-2/IDAC and AML-2/ARC. We then conducted DNA microarray analysis to compare the AML-2/IDAC cells with parental AML-2/WT cells. The results of the microarray analysis revealed a severe defect in the expression of deoxycytidine kinase (dCK), which plays a key role in the transformation of Ara-C to the active form in AML-2/IDAC cells. A similar event was observed in AML-2/ARC cells, but not in Ara-C sensitive AML-2/IDA cells that were resistant to idarubicin. The decreased expression of dCK also resulted in lower activity in both Ara-C resistant variants. However, no significant difference in the intracellular concentration of Ara-C was observed among the cells tested, which indicates that the Ara-C resistant phenotype in our models occurred due to the lower expression and activity of dCK rather than a change in the ability to take up Ara-C. Additionally, in vitro assays using BM cells from AML patients revealed that the expression of dCK and the sensitivity to Ara-C were correlated. Taken together, these findings demonstrate that dCK can regulate the in vitro cellular response to Ara-C in AML cells. PMID- 19287977 TI - Fluorescence diagnosis and flat lesions of urothelium: new challenges for pathologists and urologists (Review). AB - Bladder cancer is a common malignancy. Recurrence rate and progression vary greatly depending on factors such as tumor multiplicity, size, previous recurrence rates, tumor stage, tumor grade and the presence of carcinoma in situ. Treatment is expensive, recent studies demonstrated that superficial bladder cancer is a major economic burden. It is necessary to establish new kinds of techniques to improve diagnosis, therapy and follow-up, such as fluorescence diagnosis, without adding significant risk of complications. As urologists have a better sight of bladder lesions with fluorescence diagnosis, pathologists will be asked in the future to evaluate more frequently flat lesions, which up to now would not have been a matter of concern. For several reasons it is very important to have accurate and precise definitions of these flat lesions. First to permit uniform treatment of large groups of patients and second to see in large cohorts the evolution and natural history of several flat lesions, not always well known up to now. The aim of the study was to review the most important flat lesions, to demonstrate the difficulty of classifying several lesions, to introduce to urologists the new problems linked to FD and to suggest new models for accurate analysis. How far can we go in our answer as pathologists and how will it change the patient management? PMID- 19287978 TI - Breast cancer as a mitochondrial disorder (Review). AB - Mitochondria have been implicated in cell transformation since Otto Warburg considered 'respiration damage' to be a pivotal feature of cancer cells. Numerous somatic mitochondrial DNA (mtDNA) mutations have been found in various types of neoplasms, including breast cancer. Establishing the mtDNA mutation pattern in breast cancer cells may enhance the specificity of cancer diagnostics, detection and prediction of cancer growth rate and/or patients' outcomes; and therefore be used as a new molecular cancer bio-marker. The aim of this review is to summarize data on mtDNA mutation involvement in breast cancer and estimate effects of resulting amino acid changes on mitochondrial protein function. In this article published mtDNA mutation analyses are critically evaluated and interpreted in the functional context. PMID- 19287979 TI - Expression of the prolyl isomerase Pin1 is a useful indicator of sensitivity to chemoradiotherapy in advanced esophageal squamous cell carcinoma. AB - Cell cycle regulators, such as cyclinD1 and p53, play major roles in the tumor response to radiation and chemotherapy in esophageal squamous cell carcinoma (SCC). Pin1-mediated prolyl-isomerization potentiates cell cycle progression and cell proliferation, including the regulation of cyclinD1 and p53. Herein, we investigated the effect of Pin1 in association with cyclinD1 and p53 on the sensitivity of esophageal SCC to chemoradiotherapy (CRT). The expression levels of Pin1, cyclinD1 and p53 were examined immunohistochemically in endoscopic biopsy specimens from 68 advanced esophageal SCC patients before CRT to determine whether their expression levels predicted the clinical effectiveness of CRT in individual cancers. Forty-six of the 68 patients (67.6%) had an effective response to CRT, whereas 22 patients (32.4%) had an ineffective response. There was no significant correlation between clinical responses and expression levels of cyclinD1 or p53. However, the clinical response of the high Pin1 expression group was significantly higher than that of the low expression group (P=0.0200). Moreover, our data indicate that the combined immunohistochemical evaluation of Pin1, cyclinD1 and p53 expression in pretreatment biopsy samples is a useful indicator of sensitivity to CRT in advanced esophageal SCC. Thus, Pin1 may influence cyclinD1 and p53 functions and predict CRT sensitivity. PMID- 19287980 TI - A study on the mechanism of 5-aminolevulinic acid photodynamic therapy in vitro and in vivo in cervical cancer. AB - 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is a new ablation treatment for tumors, while its function mechanism in cervical cancer has not been elucidated. In this study, we investigated the effects of ALA-PDT on the cervical cancer cell line Me180, to search for optimal parameters of PDT. ALA-PDT on proliferation of Me180 was examined by MTT assay to find the optimal function parameters of ALA-PDT. Apoptosis was observed by using AnnexinV-FITC/PI double staining, Hoechst 33342 staining and May-Grunwald-Farbstoff Giemsa staining. Furthermore, we established a tumor model and 6 mice of each group underwent measurement of the tumor size on days 3, 7, 14, and 21 after treatment. The mRNA expression of survivin, bcl-2, p53, bax and bad in Me180 cells were detected by real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) in vitro and in vivo. Finally, we compared the effects between topical and intravenous administration. Based on the above studies, we found ALA-PDT induced apoptosis and G0-G1 phase arrest of Me180 cells. The tumor volume of the topical administration and PDT group was the smallest at 7-14 days post-PDT. H&E staining showed remarkable subcutaneous necrosis in the PDT groups. The mRNA expression of survivin and bcl-2 in Me180 cells were suppressed post-PDT. Topical administration of PDT is recommended in treating cervical cancer so as to minimize the side-effects and inconvenience of phototoxic reaction brought by PDT. Our data may contribute to making the mechanism of PDT on cervical cancer clearer and give some useful suggestions for clinical application. PMID- 19287981 TI - Inhibitory effect of sulindac on DMBA-induced hamster cheek pouch carcinogenesis and its derived cell line. AB - In order to investigate the involvement of cyclooxygenase (COX)-2 in oral carcinogenesis and chemoprevention for it, we examined the COX-2 expression during dimethylbenzanthracene (DMBA)-induced hamster cheek pouch carcinogenesis and the inhibitory effect of sulindac, a non-steroidal anti-inflammatory drug (NSAID), on the carcinogenesis and its derived squamous carcinoma cell line HCPC 1. From the beginning of DMBA application, basal diet or diets containing sulindac 200 or 400 ppm were given to hamsters, and observation of tumor development and measurement of body weight were performed. Immunohistochemical analysis revealed that COX-2 expression was increased toward carcinogenesis from epithelial dysplasia to squamous cell carcinoma (SCC). All hamsters developed SCC, but the onset of carcinoma formation was significantly delayed up to 14.8 and 11.8 weeks in the 200 ppm, and 400 ppm sulindac group, respectively, as compared to 8.7 weeks in the control group. In addition, tumor growth was retarded in the group of sulindac treatment, and mean survival time was 23.7 weeks in the control group and 36.3 and 33.8 weeks in the 200 and 400 ppm sulindac group, respectively. Body weight loss was not observed during the experimental period. Histologically, administration of sulindac inhibited angiogenesis in the tumor stroma. Treatment with sulindac sulfide, an active metabolite of sulindac, caused inhibition of cell growth, PGE2 production and VEGF production in HCPC-1 cells in vitro. Expression of COX-2 protein in HCPC-1 cells was also decreased 2-fold by treatment with sulindac sulfide. It was thus indicated that inhibitory effects were partly due to inhibition of tumor angiogenesis by sulindac. These findings suggested the involvement of COX-2 in DMBA-induced hamster cheek pouch carcinogenesis and the chemopreventive potential of sulindac. PMID- 19287982 TI - Co-expression of keratinocyte growth factor and K-sam is an independent prognostic factor in gastric carcinoma. AB - Keratinocyte growth factor (KGF) from gastric fibroblasts have been reported to stimulate proliferation of scirrhous gastric cancer cells with K-samII amplification in a paracrine manner. The aim of this study was to evaluate the clinical significance of the co-expression of K-sam and KGF in gastric carcinomas. A total of 136 primary gastric tumors were investigated by staining with antibodies against K-sam and KGF. K-sam expression on cancer cells and KGF expression on fibroblasts was estimated. The relationship between the K-sam and/or KGF expression and the clinicopathological characteristics were analyzed. K-sam expression was positive in 42 (31%) of 136 gastric carcinomas. K-sam expression was positively correlated with scirrhous cancer (p<0.001), diffuse type (p=0.031), invasion depth (p=0.018) and infiltration type (p<0.001). Prognosis of K-sam positive patients was significantly poorer than that of K-sam negative patients (p<0.001). The prognosis of patients with both K-sam and KGF positive tumors was significantly worse in comparison to either negative tumors (p<0.001). In 94 patients with a curative resection, a multivariate analysis revealed the co-expression of K-sam and KGF to be an independent prognostic factor (p=0.029). In conclusion, the co-expression of K-sam and KGF in gastric cancer might be a useful prognostic factor. PMID- 19287983 TI - A preoperative evaluation for neo-infantile liver tumors using a three dimensional reconstruction of multidetector row CT. AB - Multidetector row CT (MDCT), which has been used extensively in adult patients, has also recently been used for the evaluation of children. As pediatric surgeons, we preoperatively examined 10 cases of liver tumors by MDCT and performed three-dimensional reconstruction and a volumetric analysis. Instead of angiography, which requires general anesthesia in children, this method can provide a fine image of the anatomy between the tumor and the vessels, as well as identify the presence of any anomalous vascular branches. It also makes it possible to calculate the residual liver volume for the proposed operation and to determine the optimal cut line. However, there are still certain problems associated with pediatric cases, including the determination of the appropriate volume of contrast medium, the occurrence of allergic reactions to the contrast medium, and the timing of enhancement. The resolution of the specific problems in the pediatric application of MDCT, and the development of a more effective procedure is thus required. PMID- 19287984 TI - Malignant melanoma with a rhabdoid phenotype exhibiting numerous solid tumor masses: a case report. AB - We present a case of malignant melanoma with a rhabdoid phenotype in a 44-year old female with a quite unique and aggressive clinical course. Rhabdoid features are defined by characteristics such as sheets or solid trabeculae of neoplastic cells with large, vesicular, round to bean-shaped nuclei, prominent centrally located nucleoli, and abundant eccentric cytoplasm. Although various histological differential diagnoses were cited for the present case that showed 'rhabdoid features', most of them were excluded on the basis of the clinical history, tumor location, clinical behavior, and a broad panel of immunohistochemical stains. In the present case, the immunohistochemical findings were positive for vimentin, S 100 protein, melan-A, and EMA, but negative for HMB45, cytokeratin, CD34 and desmin. In addition, the positive expression of BAF47 was also recognized. These findings lead to the conclusion that this quite unique aggressive soft tissue tumor should therefore be diagnosed as malignant melanoma with a rhabdoid phenotype. PMID- 19287985 TI - Estrogen and progestin regulate HIF-1alpha expression in ovarian cancer cell lines via the activation of Akt signaling transduction pathway. AB - Our previous study revealed that estrogen regulates nm23-H1 expression thus promoting cell migration-invasion via activating PIK3/Akt pathway. In this study, we explored the effect of hormone on hypoxia-inducible factor-1 (HIF-1alpha), a key factor in cancer invasion and metastasis, via activation of Akt signaling transduction pathway. We treated two ovarian cancer cell lines ES-2 and SKOV3 with 17beta-estradiol, methoxyprogesterone acetate (MPA) only, or hormone combined with and Akt, MAPK pathway inhibitor, or transefected with siRNA targeting Akt sequenced with hormone. Expression of HIF-1alpha was measured by Western blotting. We observed the effect of hormone on nm23-H1 expression after the cells were transfected by siRNA targeting HIF-1alpha or treated with CoCl2 to induce HIF-1alpha overexpression. The 17beta-estradiol increased HIF-1alpha expression in ovarian cancer cells, and this upregulatory effect was abrogated by Akt inhibitor LY294002 (P<0.05) and Akt siRNA interference (P<0.05), but not affected by MAPK inhibitor PD980059 (P>0.05). MPA had the opposite effect. Nm23 H1 protein expression in ES-2 and SKOV3 cells were decreased after treatment with 17beta-estradiol (P<0.05), whereas MPA had the opposite effect. The effect was attenuated by HIF-1alpha siRNA (P<0.05) and enhanced by HIF-1alpha overexpression after CoCl2 treatment (P<0.05). Our data suggest that estrogen and progestin regulate HIF-1alpha expression via Akt signaling pathway, affecting nm23-H1 expression in influencing cell metastasis. PMID- 19287986 TI - A gastrointestinal stromal tumor presenting as a pelvic mass: A case report. AB - Gastrointestinal stromal tumors (GISTs) represent 0.1-1% of gastrointestinal malignancies. They are commonly asymptomatic and found incidentally during laparoscopy, surgical procedures or radiological studies. Diagnosis is based on histology and immunohistochemistry, while the role of imaging studies is not diagnosis-specific. We present the case of a 38-year-old patient complaining of an increase in her abdominal circumference. Consequently, a vaginal examination, a transvaginal ultrasound and an MRI of the abdomen and pelvis were carried out. It should be noted that a preoperative diagnosis of GISTs is uncommon, due to the rarity and many presentations of the disease. Ultrasound and MRI are not able to differentiate a GIST from ovarian cancer. However, if a pelvic mass is detected, the possibility of a non-gynaecological tumor has to be considered. PMID- 19287987 TI - Differential expression of osteocalcin during the metastatic progression of prostate cancer. AB - Osteocalcin expression is restricted to osteoblasts and serum osteocalcin level is elevated in metastatic bone tumors including prostate tumors, which predominantly metastasizes to the bone and causes typical osteoblastic lesions. Previously, we have reported that osteocalcin RNA is widely expressed but incompletely spliced in the prostate including prostate tumors. Considering that many studies using osteocalcin-driven gene therapy have been conducted to treat hormone refractory metastatic tumors, detailed mechanisms controlling osteocalcin expression needs to be clarified. We aim to learn how osteocalcin expression is regulated during the metastatic process of prostate cancer. We applied assays of immunohistochemistry and RNA in situ hybridization in prostate tumors acquired from prostate (15) and metastatic sites, 13 from lymph node and 14 from bone. RT PCR analysis in various cultured prostate cells was also performed. As predicted, osteocalcin RNA was highly expressed in most prostate epithelial cells of tumors, regardless of metastatic status of the tumor. However, osteocalcin protein was undetectable in tumors acquired from the primary site or lymph nodes whereas protein was highly expressed in the majority of bone-metastasized prostate tumors. RT-PCR analysis demonstrated that there was more completely spliced form of osteocalcin RNA present in bone-derived prostate cancer cells. Our data suggest that osteocalcin RNA was expressed but not completely spliced in non-bone environment, ultimately resulting in improper production of osteocalcin protein. This study explains why serum osteocalcin level is increased in patients with bone-metastasized prostate cancers. Yet, it remains to be clarified what regulates bone-specific osteocalcin RNA splicing in prostate tumors. PMID- 19287988 TI - Immunohistochemical analysis of human arrest-defective-1 expressed in cancers in vivo. AB - The arrest-defective-1 (ARD1) gene has been reported to be important in yeast cell cycle regulation, and recent studies have shown that human arrest-defective 1 (hARD1) is related to cancer cell proliferation. To investigate the expression pattern of hARD1 protein in cancer tissues, immunohistochemical analysis was performed to analyze the hARD1 expression pattern in 400 cases of 19 types of common cancer and 133 non-cancer samples from 11 tissue types. hARD1 protein was expressed extensively in cancer tissues including glandular carcinoma and squamous cancer, and the positive rate was 71.5% (15/20) in urinary bladder cancer, 62.5% (30/48) in breast cancer and 57.1% (8/14) in cervical carcinoma. The average hARD1-positive rate was 52.3% in cancers and 31.5% in non-cancers, for which the difference was significant (p<0.005). Comparing the staining intensity of different fields in the same section, the hARD1 protein was highly accumulated in cancer cells when compared to the cells adjacent to cancer. The positive rate of breast and intestinal cancer was obviously higher than corresponding non-cancers (p<0.05 and 0.01). These findings suggest that the accumulation of hARD1 protein may be related to carcinogenesis of various types of cancer. PMID- 19287989 TI - Influence of microenvironments on microcirculation patterns and tumor invasion related protein expression in melanoma. AB - This study aimed to investigate the influence of different microenvironments on melanoma microcirculation patterns, invasiveness and metastatic behavior. Sixty C57BL/6J mice were randomly divided into two groups with 30 mice per group. Melanoma B16 cells were injected into the subretinal space and groin area of mice synchronously. The number of each type of microcirculation pattern was counted. Invasion and metastasis were observed. Epithelial cell kinase (EphA2), matrix metalloproteinase (MMP)-2 and -9 expression and their mRNA levels were detected by immunohistochemical staining and real-time PCR and compared between the two groups. Five invasions and six lung metastases were found in the subretinal group while no invasion and metastasis were found in the groin group. The number of vasculogenic mimicry (VM) was significantly higher in the subretinal group (P=0.000). However, no significant difference in the numbers of mosaic and endothelium-dependent vessels was observed between the two groups (P=0.076 and 0.146, respectively). EphA2, MMP-2 and MMP-9 expression was significantly higher in the subretinal group. The mRNA levels of EphA2, MMP-2 and MMP-9 were slightly higher in the subretinal tumors (P=0.002, 0.001 and 0.001, respectively). In conclusion, this experimental paradigm can be a powerful one in which to investigate tumor-microenvironment interactions in melanoma. Tumor cells in the intraocular microenvironment had increased EphA2 expression which induced the formation of VM channels. Moreover, expression of MMP-2 and -9 in tumor tissue was increased to enhance the invasiveness and metastatic behavior. PMID- 19287990 TI - Overexpression of the LKB1 gene inhibits lung carcinoma cell proliferation partly through degradation of c-myc protein. AB - LKB1 encodes a serine/threonine kinase generally inactivated in human lung cancers, which mediates cancer cell proliferation, migration and differentiation, but its biological function has not been completely elucidated. In this study, we demonstrated that LKB1 was associated with a substantial reduction of c-myc expression by using an inducible LKB1 expression system in the LKB1-null lung cell line A549. Nevertheless, the reduction of the c-Myc gene expression was not accompanied by corresponding reduction of mRNAs but protein, which can be abrogated by a proteosome inhibitor (MG132), suggesting that the reduction was associated with their increased degradation rather than transcriptional controls. Our results implied that the expression of c-Myc protein decreased by LKB1 in transfected cells may be a contributory factor in the process of cell proliferation. Overexpression of the LKB1 gene could inhibit the activation of ERK1/2 and STAT3 signaling pathways involved in the cell proliferation. Thus, LKB1-induced functional operation on c-Myc in promoting cell proliferation may occur in a novel mechanism, which may be regulated by ERK1/2 and/or STAT3 signal pathways in human lung carcinoma cells. Furthermore, our results give some insights into the understanding of how LKB1 inactivation contributes to lung carcinogenesis and emphasizes the central role played by LKB1 in lung cancer development. PMID- 19287991 TI - Comparative study of human colonic tumor-derived endothelial cells (HCTEC) and normal colonic microvascular endothelial cells (HCMEC): Hypoxia-induced sVEGFR-1 and sVEGFR-2 levels. AB - Colorectal carcinoma growth and progression is dependent on the vasculature of the tumor microenvironment. Tumor-derived endothelial cells differ functionally from their normal counterpart. For this reason we isolated microvascular endothelial cells from human colon cancer tissue (HCTEC) and compared them with endothelial cells from normal colonic tissue (HCMEC) of the same donor. Since hypoxia is a universal hallmark of carcinomas, we examined its effects on HCTEC of five patients in comparison with the corresponding HCMEC, with respect to the secretion of the soluble form of the two important vascular endothelial growth factor (VEGF) receptors, VEGFR-1 and -2. After dissociation by dispase/collagenase of central non-necrotic tumor areas obtained from colon carcinomas, HCTEC were isolated using CD31-coated magnetic beads and cultivated as monolayers. Subsequent characterization studies demonstrated the endothelial phenotype, including VEGFR-1 and -2 mRNA and protein expression as well as E selectin expression, up-regulated after LPS, TNFalpha and IL-1beta stimulation. sVEGFR expression analyses were performed using ELISA. In comparison with HCMEC markedly lower sVEGFR-1 protein concentrations were found in HCTEC. These low sVEGFR-1 levels remain unchanged under hypoxia. In contrast, sVEGFR-2 was significantly decreased in both HCMEC and HCTEC under hypoxic conditions (p45% of A549 and H446 cells formed large clones that are able to generate subclones and subsequently give rise to xenograft tumors in the BALB/C-nude mouse. Both CD133+ and CD133- cells of A549 and H446 possessed stem cell characteristics and exposure to Hoechst 33342 affected the clonogenicity and proliferation of single A549 and H446 cell. Moreover, based on cell cycle phase distribution and growth curve, most A549 and H446 cells may not be stem cells, but they acquired stem cell characteristics when they were alone. Therefore, these cells that formed large clones should be called cancer initiating cells, not stem cells. Thus, we concluded that >45% of A549 and H446 cells were cancer initiating cells, although many of them were neither CD133+ cells nor side population cells. PMID- 19288000 TI - Mutational analysis of Smad7 in human cervical cancer. AB - Our aim was to carry out mutational analysis of Smad7 exon 4 that codes for the Smad7 MH2 domain in human cervical cancer tissues. This study is warranted since genetic abnormalities of components of the transforming growth factor-beta (TGF beta)/Smad signaling system have been implicated in a variety of human cancers. Further, Smad7 is an important member of the Smad family that functions as a negative feedback regulator of TGF-beta responses. By direct sequencing, we screened 60 histopathologically confirmed human cervical carcinomas for mutations in exon 4 of Smad7 that encodes the MH2 domain of the protein, a region of the protein believed to be critical for receptor interaction. No mutations or aberrations could be identified in any of the 60 analyzed tumor samples. However, we identified a previously reported, heterozygous, silent G to C variant in codon 391 of Smad7 that was found twice in the set of 60 clinical samples. Thirty of the clinical samples analyzed in the study were positive for the presence of high risk human papillomavirus (HPV) subtypes, while the rest were oncogenic HPV negative. The two instances of the silent variant was found in oncogenic HPV negative samples. We report mutational analysis of Smad7 in cervical cancer for the first time. Mutations and variants of Smad7 are unlikely to be of major significance to the pathogenesis of HPV-induced cervical cancer. PMID- 19288001 TI - Phase I study of combination chemotherapy with irinotecan hydrochloride and nedaplatin for cervical squamous cell carcinoma: Japanese gynecologic oncology group study. AB - The aim of this study (JGOG1063) was to determine the recommended dose (RD) for combination chemotherapy with irinotecan hydrochloride (CPT-11) and nedaplatin (NDP) for advanced cervical squamous cell carcinoma. CPT-11 was given intravenously in fixed doses of 60 mg/m2 on days 1 and 8 and NDP, in escalating doses, on day 1, every 4 weeks. A total of 15 patients were enrolled in the study. At level 1 (NDP: 50 mg/m2), one of the 3 patients developed grade 3 diarrhea, so 3 additional patients were enrolled at this level. As none of the 3 additional patients exhibited dose-limiting toxicity, level 1 was elevated to level 2 (NDP: 60 mg/m2). The maximum tolerated dose was not reached, even at the highest dose level (level 4; NDP: 80 mg/m2). No further dose escalation was carried out, and level 4 (CPT-11: 60 mg/m2, NDP: 80 mg/m2) was determined as the RD. PMID- 19288002 TI - Beta-interferon, retinoids and tamoxifen in metastatic breast cancer: long-term follow-up of a phase II study. AB - Based on a series of in vitro data, including the additive and/or synergistic antiproliferative effect of interferon and tamoxifen on breast cancer cell lines, and on clinical reports, we designed a pilot phase II study to test the activity and toxicity of simultaneous administration of beta-interferon (beta-IFN), retinoids (R) and tamoxifen (TAM) as a salvage therapy in a group of patients with metastatic breast cancer (MBC). Herein we describe the outcome of this cohort of patients after a median follow-up of 150 months. Sixty-five stage IV breast cancer patients, 13 pre-treated with hormones, 38 with chemotherapy and 15 with both, received, as a salvage therapy, TAM, beta-IFN and R. Among 65 evaluable patients, 36 achieved a clinical response (55.5%) (95% c.i. 42-67.7%). Toxicity was moderate and mainly hepatic. Median progression-free and overall survival, which did not show any statistically significant difference in patients with different estrogen and progesterone receptor content, were 43 months and 47.9 months, respectively. In conclusion, the study shows that long-term treatment with TAM, beta-IFN and R in MBC is feasible, has moderate toxicity and seems to give a long-term benefit, irrespective of the receptorial status. PMID- 19288003 TI - Novel cell culture model for prevention of carcinogenic risk in familial adenomatous polyposis syndrome. AB - Clinical familial adenomatous polyposis (FAP) syndrome represents a high risk pre invasive precursor for colon cancer, and is characterized by germ line mutation in the adenomatous polyposis coli (APC) tumor suppressor gene. Cellular models with relevant genetic and biological characteristics should provide important mechanistic leads for predisposition and preventive intervention. Cloned colon epithelial cell line from the Apc850 Min/+ mouse represented a model for FAP. Cell cycle progression, cellular apoptosis and anchorage-independent growth represented the biomarkers for carcinogenic risk. The Apc mutant 850Min COL-Cl1 cells exhibited decreased G0/G1:S+G2/M ratio, increased S+G2/M:subG0 ratio, and increased anchorage-independent colony formation, indicating loss of homeostatic growth control and gain of anchorage-independent growth. Growth of these cells in serum-depleted medium was promoted by mitogenic insulin and epidermal growth factor, and inhibited by anti-mitogenic transforming growth factor-beta1 and dexamethasone. Treatment with low dose combinations of synthetic enzyme inhibitor difluoro methylornithine (DFMO), synthetic non-steroidal anti-inflammatory drug sulindac (SUL), and naturally occurring epigallocatechin gallate (EGCG), and eicosapen-taenoic acid (EPA) produced cytostatic growth arrest and inhibited anchorage-independent colony formation. These data identify a novel cell culture model and validate a mechanism-based approach to prioritize combinations of effective chemopreventive compounds for prevention/therapy of colon cancer. PMID- 19288004 TI - Correlation between efficacy and skin rash occurrence following treatment with the epidermal growth factor receptor inhibitor cetuximab: a single institution retrospective analysis. AB - Several trials show a relationship between skin toxicity, response rate, and overall survival in cetuximab-treated patients. We analyzed our database to evaluate the importance of skin rash as a surrogate marker of favorable outcome in cancer patients referred to our institution in the last three years. We retrospectively analyzed 90 cetuximab-treated patients: 57 colon cancer patients, 10 NSCLC patients, 14 locally advanced esophageal cancer patients, and 9 miscellaneous. A significant correlation was observed between skin rash and response to therapy. Skin rash was experienced by 93% of PR and 100% of CR patients. The mean TTP was 184 days in patients showing skin rash and 94 days in patients without skin rash, respectively. On multivariate analysis, skin rash was demonstrated to be the only independent prognostic variable with regard to TTP. Patients who did not develop skin rash had a 2-fold greater likelihood to manifest tumor progression significantly earlier than patients who developed skin rash. In our series, a statistically significant correlation between rash, response rate, and TTP was demonstrated in 90 cetuximab-treated patients. Skin toxicity was confirmed as the only clinical variable able to predict the response to cetuximab. PMID- 19288005 TI - Bone marrow stem cell damage after three different chemotherapy regimens for advanced Hodgkin's lymphoma. AB - The aim of this study was to evaluate the apoptotic damage to bone marrow cells caused by three chemotherapy regimens for advanced Hodgkin's lymphoma, ABVD, COPPEBVCAD and BEACOPP, which were randomly administered in the HD 2000 GISL trial. Bone marrow mononuclear cells (BMMCs) stained with anti-CD34 antibody and Annexin V, were evaluated by flow cytometry before starting chemotherapy, 30 days after completing chemotherapy and after 6 months. Results are expressed as the percentages of BMMCs positive to anti-CD34, to Annexin V or to both. Fourteen patients treated with ABVD, 11 with COPPEBVCAD and 13 with BEACOPP were evaluated before and 30 days after treatment. Late assessments were made in 6, 7 and 8 of them, respectively. No differences were found among the pretherapeutic flow cytometry findings in relation to the staging characteristics (marrow involvement included). All the regimens increased the apoptotic fraction of the whole mononuclear bone marrow cells (COPPEBVCAD did so significantly) and increased the CD34+ compartment (with significant early differences after ABVD and BEACOPP, tending to late persistence for ABVD, only). All the regimens increased the apoptotic CD34+ cells within the whole BMMC population (significantly after BEACOPP), although with a general trend to decrease in their percentage within the CD34+ compartment over time, even after the most dose-dense regimens. Based on the variations induced in the apoptotic fraction of all mononuclear and CD34+ cells, ABVD was the least toxic regimen and COPPEBVCAD the most toxic one. PMID- 19288006 TI - Expression of 5-fluorouracil-related enzymes in lung cancer: ELISA characterizes enzyme activity and messenger RNA expression. AB - The enzymes thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD) and orotate phosphoribosyl transferase (OPRT) are involved in the metabolism of the anticancer drug 5-fluorouracil (FU). Expression of TS, DPD and OPRT in cancer tissue has been reported to be associated with sensitivity and/or resistance to 5 FU therapy. However, the role of TS, DPD and OPRT expression in lung cancer has not been fully established. Furthermore, among several measuring methods, it is not clear which method effectively predicts the response to 5-FU therapy. The aim of this study was to analyze the expression of 5-FU-related enzymes using enzyme linked immunosorbent assay (ELISA) and to examine the correlation of ELISA and the results obtained using different measuring methods such as reverse transcript polymerase chain reaction (RT-PCR), immunohistochemistry, and enzymatic activity. Lung cancer specimens were obtained from 134 patients who underwent curative resection for lung cancer. As a pilot study, enzyme expression of 11 samples was measured using 4 different methods for DPD: RT-PCR, immunohistochemistry, enzymatic activity and ELISA. The relationships between pairs of results were compared, and then enzyme protein expression was measured using ELISA in 119 patients with adenocarcinoma. Of the 4 independent methods, the highest correlation was observed between protein expression measured by ELISA and enzyme activity. The correlation of gene expression and ELISA was also significant. The protein level in stage I adenocarcinoma measured using ELISA was 13.0+/-24.8 ng/mg protein for TS, 362.2+/-264.3 ng/mg protein for DPD and 4.5+/-2.0 ng/mg protein for OPRT. The predictive value of the enzymes for prognosis and the effectiveness of 5-FU was not determined as few recurrences were observed during the short follow-up period. In conclusion, ELISA is a simple and reliable method to measure key enzymes related to 5-FU therapy. PMID- 19288007 TI - Seventeen years of experience with ATRA-based therapy for acute promyelocytic leukaemia: long-term follow-up of patients treated at S. Martino Hospital, Genoa. AB - We conducted a long-term follow-up retrospective study on 91 consecutive newly diagnosed acute promyelocytic leukaemia (APL) patients. Induction and consolidation therapy were well-tolerated by most patients. Of the 79 patients who were initially treated with the all-trans retinoic acid (ATRA)-containing regimens, there were 3 haemorrhagic deaths during the first period of therapy (4%) and one in consolidation which was due to infection. Following consolidation, molecular assessment of response was performed on 67 patients, and 66 were found to have achieved cytogenetic and molecular remission (98%). After a median follow-up of 100 months (12-192), 10 of the 75 patients who achieved complete remission (13%) relapsed. Seventy-eight percent of the patients were expected to be alive at 14 years from diagnosis, i.e., 90 and 48% of patients of intermediate-low risk and high risk at presentation, respectively (p=0.0009). Sixty-nine patients were in molecular remission after first-line and/or salvage therapy (74%). To date, 4 patients out of the 91 have undergone salvage allogeneic transplant (4%). PMID- 19288008 TI - The role of Id-1 in chemosensitivity and epirubicin-induced apoptosis in bladder cancer cells. AB - Recurrence and progression are the major problems in the treatment of bladder cancer. Increased expression of Id-1, a basic helix-loop-helix transcription factor, has recently been shown in several types of advanced cancer. Some studies have provided evidence to suggest that Id-1 can be considered a potential therapeutic target. The objective of this study was to investigate the role of Id 1 in the chemosensitivity of bladder cancer cells, and the effect of Id-1 on chemotherapeutic drug-induced apoptosis in bladder cancer cells. We compared the different sensitivity to epirubicin in RT112 and MGH-U1 cell lines with different Id-1 expression. Then, we transfected different vectors into RT112 and MGH-U1 respectively, and generated the stable Id-1 up-regulation and down-regulation transfectants. The results of cell viability assay showed up-regulation of Id-1 in RT112 leading to increased sensitivity in response to epirubicin, and down regulation of Id-1 increased cellular sensitivity to epirubicin. Furthermore, the analysis of apoptosis related protein revealed that up-regulation of Id-1 suppressed epirubicin-induced apoptosis and down-regulation of Id-1 leading to increased epirubicin-induced apoptosis. Wound closure assay showed up-regulation of Id-1 leading to improved migration abilities of bladder cancer cells under chemotherapy. Our results suggest that up-regulation of Id-1 in bladder cancer cells lead to increased cell viability in response to epirubicin by its improved anti-apoptotic role, and down-regulation of Id-1 increases cellular sensitivity to epirubicin by increased anticancer drug-induced apoptosis. PMID- 19288009 TI - Clinical comparison of QOL and adverse events during postoperative adjuvant chemotherapy in outpatients with node-positive colorectal cancer or gastric cancer. AB - The aim of this study was to evaluate the quality of life (QOL) from the performance status (PS) and face scale (FS), and to compare adverse events (AEs) during chemotherapy in 28 patients with node-positive colorectal cancer (NP-CRC) and 15 patients with node-positive gastric cancer (NP-GC). The anticancer regimen consisted of 5-FU/LV+CPT-11 for NP-CRC and 5-FU+low-dose CDDP for NP-GC. Results were evaluated after completion of three courses. QOL evaluation revealed no significant differences between the two groups with respect to PS and FS. Among hematological AEs, grade 1/2 mild leucopenia was significantly more common in NP CRC than NP-GC patients (p<0.05), while grade 1/2 mild thrombocytopenia was significantly more common in NP-GC than NP-CRC patients (p<0.05). Among non hematological AEs, grade 1/2 mild neuropathy (olfactory nerve) was significantly more common in NP-CRC than NP-GC patients (p<0.05). The monthly cost for one course was approximately euro586.8 for NP-CRC patients and approximately euro181.8 for NP-GC patients. These results suggest that first-line postoperative outpatient adjuvant chemotherapy for NP-CRC and NP-GC shows no significant differences with respect to QOL, but both AEs and the cost are higher for NP-CRC than for NP-GC. PMID- 19288010 TI - Methylation of CLDN6, FBN2, RBP1, RBP4, TFPI2, and TMEFF2 in esophageal squamous cell carcinoma. AB - In the development and progression of cancer, tumor suppressor genes may be silenced by mechanisms such as methylation. Thus the discovery of new genes silenced by methylation may uncover new tumor suppressor genes, and improve our understanding of cancer biology. In this study we investigated the methylation of 19 genes in esophageal squamous cell carcinoma. Methylation was measured in 10 of these genes in esophageal squamous cell carcinoma cell lines: CDH13, CLDN6, C16orf62, FBN2, FNBP1, ID4, RBP1, RBP4, TFPI2 and TMEFF2. To determine if there was a correlation between DNA methylation and gene silencing, each cell line was cultured with or without the demethylating drug 5-aza-2'-deoxycytidine (aza-dC). For 6 genes (CLDN6, FBN2, RBP1, RBP4, TFPI2 and TMEFF2) there was an association between reduction of methylation and increase in mRNA expression in the demethylated cell lines. The frequency of the methylation of these 6 genes in esophageal squamous cell carcinoma resection specimens was also investigated. All 6 genes showed more frequent methylation in the tumor than the matched proximal resection margin of uninvolved esophagus. There was a significant difference in the frequency of methylation and in the extent of the methylation between the cancer and the margin tissues for CLDN6, FBN2, TFPI2 and TMEFF2 (P=0.0007, P=0.0048 P=0.0002 and P<0.0001, respectively). This is the first report of gene silencing by methylation of CLDN6, FBN2, RBP4, TFPI2 and TMEFF2 in esophageal squamous cell carcinoma. PMID- 19288011 TI - Tanshinone IIA inhibits leukemia THP-1 cell growth by induction of apoptosis. AB - Tanshinone IIA, a diterpene quinone extracted from the traditional herbal medicine, Salvia miltiorrhiza Bunge, has been reported to have anti-tumor effects on a large variety of cancer cells. The present study was undertaken to investigate the in vitro antiproliferation and apoptosis inducing effects of Tanshinone IIA on leukemia THP-1 cell lines and its mechanisms of action. MTT assay was used to detect the cell growth inhibitory rate; cell apoptotic rate and the mitochondrial membrane potential (Deltapsim) were investigated by flow cytometry (FCM), apoptotic morphology was observed by Hoechst 33258 staining and DNA fragmentation analysis. The expression of caspase-3 and different apoptosis modulators were analyzed by Western blotting. The results revealed that Tanshinone IIA inhibited the growth of THP-1 cells and caused significant apoptosis, the suppression was both in time- and dose-dependent manner. After treatment by Tanshinone IIA for 48 h, the percentage of disruption of Deltapsim gradually increased in a dose-dependent manner along with marked changes of cell apoptosis. Western blotting showed cleavage of the caspase-3 zymogen protein (32 kDa) with the appearance of its 20-kDa subunit and a dose-dependent cleavage of PARP, with the appearance of 89-kDa fragment; The expression of Bcl-2 and survivin was down-regulated remarkably while Bax expression was up-regulated concurrently after the cells were treated with Tanshinone IIA for 48 h. We therefore conclude that Tanshinone IIA has significant growth inhibition effects on THP-1 cells by induction of apoptosis, and that Tanshinone IIA-induced apoptosis on THP-1 cells is mainly related to the disruption of Deltapsim and activation of caspase-3 as well as down-regulation of anti-apoptotic protein Bcl 2, survivin and up-regulation of pro-apoptotic protein Bax. The results indicate that Tanshinone IIA may serve as a potential anti-leukemia reagent. PMID- 19288012 TI - Enhanced effects of PPARgamma ligands and RXR selective retinoids in combination to inhibit migration and invasiveness in cancer cells. AB - Experimental data from in vitro and in vivo models indicate that peroxisome proliferator-activated receptor (PPAR) ligand activation regulates differentiation and induces cell growth arrest and apoptosis in a variety of cancer types. Thiazolidinediones such as ciglitazone (CGZ) constitute the most well-known synthetic ligands for PPARgamma. We previously reported a remarkable antitumor effect of the retinoid 6-OH-11-O-hydroxyphenantrene (IIF), synthetic retinoid X receptors (RXRs) agonist, on many cancer cell lines. Since PPARs bind to DNA as heterodimers with RXRs, in this study we investigated if IIF potentiates the antitumoral properties of the PPARgamma ligand CGZ in glioblastoma U87MG and melanoma G361 cells. Our results show that either IIF or CGZ inhibited cell growth and tissue invasion ability, but these properties were enhanced by using IIF and CGZ in combined treatment. Since matrix metalloproteinases (MMPs) play a major role in tumor cell invasion, we analyzed the effect of IIF and CGZ on MMP2 and MMP9 activity and expression. The addition of IIF to CGZ resulted in a decrease of MMP2 and MMP9 expression and activity, higher than when each agent was used alone. Furthermore, treatment with IIF and/or CGZ enhanced PPARgamma expression but both agents in combined treatment caused the maximum efficiency. Finally, we demonstrated that IIF can potentiate PPARgamma trascriptional activity induced by CGZ, by evaluation of peroxisome proliferator-responsive element transactivation. In conclusion, these findings suggest that the RXR selective retinoid IIF, in combination with the PPARgamma ligand CGZ, may provide a therapeutic advantage in cancer treatment. PMID- 19288013 TI - Pro-inflammatory cytokine release by peripheral blood mononuclear cells from patients with advanced pancreatic cancer: relationship to acute phase response and survival. AB - The acute phase protein response (APPR) and peripheral blood mononuclear cell derived inflammatory cytokine production was assessed in patients with advanced pancreatic cancer and age-matched healthy volunteers. We examined the relationship between the APPR, cytokine production and survival in these patients. Forty-two patients with pancreatic cancer cachexia and twelve age matched healthy controls were recruited. The nutritional status, Karnofsky performance score, C reactive protein (CRP), serum interleukin-6, and in vitro monocyte interleukin-1 and interleukin-6 production were measured. The dates of death of the pancreatic cancer patients were subsequently obtained and appropriate patient variables at baseline were entered into a Cox's proportional hazards model. The cancer patients had significantly lower: body mass index, Karnofsky performance score, serum albumin and elevated CRP and stimulated interleukin-6 production. Both univariate and multivariate analysis demonstrated a strong association between tumour stage, CRP, stimulated interleukin-6 production and survival. Monocytes in cachectic pancreatic cancer patients are primed to produce high levels of interleukin-6 when stimulated. Overproduction of interleukin-6 has a negative impact on survival. Decreased survival is associated with an elevated APPR. While the elevated APPR is probably related to locally produced interleukin-6 in the liver, it seems possible that locally and systemically produced interleukin-6 influences survival. PMID- 19288014 TI - Pin1 overexpression is associated with poor differentiation and survival in oral squamous cell carcinoma. AB - Phosphorylation on certain Ser/Thr-Pro motifs is a major oncogenic mechanism. The conformation and function of phosphorylated Ser/Thr-Pro motifs are further regulated by the prolyl isomerase Pin1. Pin1 has been shown to be prevalently overexpressed in human breast cancer cell lines and cancer tissues and to play a critical role in the transformation of mammary epithelial cells by activating multiple oncogenic pathways. Pin1 expression was found to be an excellent independent prognostic marker in prostate cancer. However, little is known about Pin1 and its downstream targets beta-catenin and cyclin D1 expressions in human oral cancers. In the present study, we quantified Pin1 expression in 74 paired normal/tumor human oral cancer samples as well as oral cancer cell lines. Pin1 was overexpressed in oral squamous cell carcinoma (OSCC) and its level correlated with beta-catenin accumulation and cyclin D1 expression. Moreover, we examined Pin1 mRNA expression in OSCC and cancer cell lines by RT-PCR analysis. The results showed that there is concordance in the relationship between the Pin1 mRNA level and Pin1 protein expression. The up-regulation of Pin1 mRNA expression in tumor part when comparing with that in non-tumor part was in agreement with that of the Pin1 protein overexpressed in OSCC. In addition, we showed that the molecular and immunohistochemical profiles of Pin1 overexpression were associated with progression of OSCC. Taken together, these results indicate that Pin1 is a regulator of cyclin D1 expression in OSCC and might play a role in oral oncogenesis. The overexpression of Pin1 can be used as an indicator for pathological diagnosis of OSCC. PMID- 19288015 TI - Decrease in malonyl-CoA and its background metabolic alterations in murine model of cancer cachexia. AB - The alterations of enzymatic activities involved in lipid degradation in cancer cachexia have not been fully elucidated. One of the two subclones of colon 26 adenocarcinoma, clone 20, with a potent ability to induce cachexia, or clone 5, without such an activity, was transplanted in to CDF-1 male mice. Murine livers were extirpated for analyses on the 14th day after tumor inoculation. The body weights and food intake of mice bearing clone 20 were all significantly lower than those of non-tumor bearing mice and mice bearing the clone 5 tumor. The decline of body weight was accompanied by a shrinkage of epididymal fat pads. Expression of spermidine/spermine N-1 acetyl transferase (SSAT) assessed by real time PCR was significantly increased in cachectic mice. Conversely, acetyl-CoA carboxylase (ACC) measured by Western blotting and malonyl-CoA levels determined by malonyl-CoA:acetyl-CoA cycling procedures were decreased in cachectic mice. Indomethacin in drinking water reversed the clone 20 induced decrease in body and fat weight and food intake, and simultaneously negated the clone 20 induced increase of SSAT expressions and decrease of ACC and malonyl-CoA amounts. Because malonyl-CoA inhibits the rate-limiting step in the beta-oxidation of fatty acids, the decreased malonyl-CoA and the background metabolic alterations may contribute to the accelerated lipolysis of cancer cachexia. PMID- 19288016 TI - Role of CCL5 in invasion, proliferation and proportion of CD44+/CD24- phenotype of MCF-7 cells and correlation of CCL5 and CCR5 expression with breast cancer progression. AB - This study was undertaken to observe the effects and possible mechanism of CC chemokine ligand 5 (CCL5) on invasion, proliferation and percentage of CD44+/CD24 subpopulation of human breast cancer line MCF-7 and to investigate the correlation of expression levels of CCL5 and its receptors with the progression of breast cancer. We used real-time RT-PCR to detect the expression levels of CCL5 and its receptors CCR5, CCR1 and CCR3 in 36 breast cancer specimens of different TNM stage and their corresponding normal breast tissue. CCL5 expression and invasive ability of four human breast cancer cell lines MCF-7, SK-BR-3, T-47D and MDA-MB-231 were analyzed by real-time RT-PCR and cell invasion assay, respectively. Effects of recombinant human CCL5 (rhCCL5) on cell proliferation and percentage of the CD44+/CD24- subpopulation in MCF-7 cells were analyzed respectively by MTT assay and flow cytometry. We also used cell invasion assay to detect the invasive ability of both CD44+/CD24- and CD44+/CD24+ subpopulations of MCF-7 cells treated with rhCCL5 and/or CCR5 monoclonal antibody. Our results revealed that CCL5 and CCR5 expression were higher in breast cancer tissue than those in their corresponding normal tissue and breast cancer tissue with higher TNM stage contained more CCL5 mRNA. In addition, CCR5 expression and invasive ability of CD44+/CD24- subpopulation were higher than those of CD44+/CD24+ subpopulation of MCF-7 cells. Moreover, treatment of rhCCL5 increased the proportion of CD44+/CD24- cells and the proliferation of MCF-7 cells. Induction of rhCCL5 increased the cell invasive ability of both CD44+/CD24- and CD44+/CD24+ cells, which could be partially antagonized by CCR5 monoclonal antibody. Collectively, our data show that CCL5 increased the proportion of CD44+/CD24- subpopulation and induced invasion and proliferation of MCF-7 cells, and expression of CCL5 and CCR5 in breast cancer tissue was positively correlated with breast cancer progression. PMID- 19288017 TI - Prognostic significance of altered expression of SDC2 and CYR61 in esophageal squamous cell carcinoma. AB - The transforming growth factor beta (TGF-beta) signaling pathway plays an important role in growth and development, and is critically involved in the genesis and development of tumors. Syndecan-2 (SDC2) and Cysteine-rich 61 (CYR61) are important genes in this pathway and SDC2 is known to be a significant upstream regulator of TGF-beta signaling. However, the roles of SDC2 and CYR61 in the development of esophageal squamous cell carcinoma (ESCC) remain unclear. In the present study, we investigated the relationship between SDC2 and CYR61 mRNA expression levels and disease prognosis in patients with ESCC. The mRNA expression of SDC2 and CYR61 was detected by quantitative real-time RT-PCR in 77 tissue specimens. Quantitative real-time RT-PCR showed that SDC2 and CYR61 mRNA expression levels were aberrant in ESCC tissue (P<0.01) and that SDC2 mRNA expression was significantly associated with tumor size (P=0.024) in ESCC. CYR61 mRNA expression was significantly associated with regional lymph node metastasis (P=0.034) and tumor size (P=0.03). A positive correlation between SDC2 and CYR61 (r=0.770; P<0.001) mRNA expression was observed. Moreover, we observed significant associations between altered expression of SDC2/CYR61 and regional lymph node metastasis (P=0.009) and TNM stages (P=0.033). Aberrant mRNA expression of CYR61 and SDC2/CYR61 (P=0.005 and P=0.026, respectively) were significantly associated with patient survival time. The multivariate Cox regression analysis showed that SDC2 and CYR61 were independent prognostic factors for survival. Our findings suggest that SDC2 may act as an a upstream regulator of the TGF-beta signaling pathway and regulate the expression of downstream target genes. Moreover, SDC2 and CYR61 expression affect the severity of cancer, and the survival of patients with ESCC. Importantly, we report that SDC2 and CYR61 are significant, independent prognostic factors for survival in ESCC. These findings may have implications for targeted therapies in patients with ESCC. PMID- 19288018 TI - Pathogenetic mechanisms and treatment options for ophthalmic pterygium: trends and perspectives (Review). AB - Pterygium is a fibrovascular lesion of the ocular surface that can display an aggressive clinical behavior and, occasionally, threaten vision. Although there is no consensus on its pathogenesis, recent evidence suggests that it is a proliferative, rather than degenerative condition strongly correlated with exposure to ultraviolet radiation (UVR) of solar light. Molecular genetic alterations reported in association with pterygium include loss of heterozygosity (LOH), point mutations of proto-oncogenes, such as K-ras and alterations in the expression of tumor suppressor genes, such as p53 or p63. Other findings in pterygium include the frequent detection of HPV DNA, ocular surface changes such as the overexpression of various proteins, including defensins and phospolipases D, as well as the up-regulation of growth factors, such as bFGF or VEGF. Although pterygium management has traditionally involved surgery, often enhanced by the use of antimetabolites, recent advancements in the understanding of molecular and biochemical events underlying pterygium pathogenesis may enable the use of less invasive treatment methods. PMID- 19288019 TI - Lecithin promotes adipocyte differentiation and hepatic lipid accumulation. AB - Lecithin is an essential biological component and widely used as a nutritional supplement for protecting cells from oxidation, increase fat burning and preventing cardiovascular disease. Lecithin contains fatty acids identified as the peroxisome proliferator-activated receptor (PPAR) agonists. However, the role of lecithin in adipogenesis and lipogenesis remains elusive. 3T3-L1 cells and mouse primary preadipocytes were used to characterize the properties of lecithin related to adipogenesis and lipogenesis. We found that lecithin promoted adipocyte differentiation and differentiation-specific gene expression, and increased triglycerides and free fatty acid levels in the adipocytes. These effects are independent of the clonal expansion of 3T3-L1 cells and the upstream PPARgamma regulator, CCAAT-enhancer-binding protein beta. Furthermore, lecithin induced lipid accumulation in human hepatoma HepG2 cells. Our data suggest that lecithin is involved in adipogenesis, lipogenesis and hepatic lipid accumulation and it is implicated in obesity and hepatic steatosis. PMID- 19288020 TI - Long-term effects of chondrospheres on cartilage lesions in an autologous chondrocyte implantation model as investigated in the SCID mouse model. AB - Microtraumata often lead to articular cartilage lesions. Due to the bradytrophic character of hyaline cartilage, these lesions are hardly repaired by the organism. Autologous chondrocyte implantation (ACI) was established for restoring isolated structural cartilage defects in knee joints. However, results are not always convincing. Human chondrocytes from patients undergoing total knee arthroplasty were cultured in monolayer followed by condensing single chondrocytes to spheroids (chondrospheres). The integrative capacity of chondrospheres was examined by implanting them into lesions in human articular cartilage specimens and co-implanting them into SCID mice. Mice were sacrificed after 4, 12 and 24 weeks. HE and safranin O staining as well as immunohistochemistry using anti-S100, anti-collagen I and II antibodies were performed and analyzed using semiquantitative scores. Integration of the chondrospheres with the (native) cartilage matrix was analyzed by determining the percentage of adhering surface. With respect to long-term stability, the chondrocytes within chondrospheres showed a typical chondrocytic morphology. Immunohistochemically, a high collagen II production was detected. Over a time period of 24 weeks, an increasing content of collagen type II, glycosaminoglycans and collagenous fibers were found. Importantly, the newly synthesized cartilaginous matrix integrated continuously with the native cartilage lesion border. In conclusion, the presented data demonstrate that chondrospheres are able to restore and conserve their phenotype for at least 24 weeks under in vivo conditions. Moreover, chondrospheres adhere to full-thickness cartilage defects and appear to produce a cartilaginous extracellular matrix which fuses with native cartilage thus generating an autologous cartilage-like repair tissue. PMID- 19288021 TI - Experimental study of millimeter wave-induced differentiation of bone marrow mesenchymal stem cells into chondrocytes. AB - Low power millimeter wave irradiation is widely used in clinical medicine. We describe the effects of this treatment on cultured mesenchymal stem cells (MSCs) and attempted to identify the underlying mechanism. Cells cultured using the whole marrow attachment culture method proliferated dispersedly or in clones. Flow cytometric analyses showed that the MSCs were CD90 positive, but negative for CD45. The negative control group (A) did not express detectable levels of Cbfa1 or Sox9 mRNA at any time point, while cells in the millimeter wave-induced groups (B and C) increasingly expressed both genes after the fourth day post induction. Statistical analysis showed that starting on the fourth day post induction, there were very significant differences in the expression of Cbfa1 and Sox9 mRNA between groups A and B as well as A and C at any given time point, between treated groups B and C after identical periods of induction, and within each treated group at different induction times. Transition electron microscopy analysis showed that the rough endoplasmic reticulum of cells in the induced groups was richer and more developed than in cells of the negative control group, and that the shape of cells shifted from long-spindle to near ellipse. Toluidine blue staining revealed heterochromia in the cytoplasm and extracellular matrix of cells in the induced groups, whereas no obvious heterochromia was observed in negative control cells. Induced cells also exhibited positive immunohistochemical staining of collagen II, in contrast to the negative controls. These results show that millimeter wave treatment successfully induced MSCs to differentiate as chondrocytes and the extent of differentiation increased with treatment duration. Our findings suggest that millimeter wave irradiation can be employed as a novel non-drug inducing method for the differentiation of MSCs into chondrocytes. PMID- 19288022 TI - Curcumin inhibits proliferation and migration by increasing the Bax to Bcl-2 ratio and decreasing NF-kappaBp65 expression in breast cancer MDA-MB-231 cells. AB - Curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione), is extracted from the plant Curcuma longa. It has cytotoxic effects and induces apoptosis in many human cancer cells but the molecular mechanisms are not fully understood. In the present study, we evaluated the effects of curcumin on human breast cancer MDA-MB-231 cells. The cytotoxic effects of curcumin on MDA-MB-231 cells were measured by MTT assay. The percentages of cell cycle were determined by flow cytometry. The protein expressions of p21, 53, Bax and Bcl-2 were examined by Western blotting. The results show that curcumin inhibits the proliferation of MDA-MB-231 cells and induces G2/M arrest in a dose-dependent manner. Curcumin increased the protein expressions of p21 and Bax, but decreased the protein expression of p53 and Bcl-2 in MDA-MB-231 cells. Our results show that one molecular mechanism of curcumin inhibits the proliferation of MDA-MB-231 cells either through up-regulating p21 expression and then inducing apoptosis, or through up-regulating the Bax to Bcl-2 ratio and then inducing apoptosis. Our results also show that curcumin inhibits the migratory activity of MDA-MB-231 cells through down-regulating the protein expression of NF-kappaBp65. Accordingly, the therapeutic potential of curcumin for breast cancer deserves further study. PMID- 19288023 TI - Changes of hepatic proteome in bile duct ligated rats with hepatic fibrosis following treatment with Yin-Chen-Hao-Tang. AB - Yin-Chen-Hao-Tang (YCHT) is recognized as a hepatoprotective agent for various types of liver diseases. Proteomics approaches were used to study hepatic and serum protein expression changes in bile duct ligated (BDL) rats following YCHT treatment for 27 days. Two-dimensional gel electrophoresis was used to analyze proteome changes. Of the proteins that exhibited changes, 17 were identified by means of matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) mass spectrometry. The major effect of YCHT was evident in cytoskeleton related protein, plectin-1. In addition, proteins involved in metabolism of lipids were also shown to be affected, including low-density lipoprotein receptor-related protein 2 precursor (glycoprotein 330) and apolipoprotein A-I precursor (ApoA-I). Significant up-regulation of keratin 8 and 19 was found in liver tissue of BDL rats. Supplementation with YCHT also triggered alterations in the above proteins. Interestingly, YCHT treatment caused a statistically significant down-regulation in the secretion of monocyte chemoattractant protein-1 (MCP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in BDL rats with fibrosis. Our results suggested that YCHT may be useful for treatment of liver fibrosis because of its possible antiapoptotic properties, and the therapeutic effects of YCHT on liver diseases might be associated with its lipid biosynthesis regulation. PMID- 19288024 TI - Genetic variants in interleukin-6 modified risk of obstructive sleep apnea syndrome. AB - Obesity and inflammation are known to correlate with the pathogenesis of obstructive sleep apnea syndrome (OSAS). Interleukin (IL)-6, an important regulator of obesity and inflammation, was reported to phenotypically increase in patients with OSAS. This study aimed to investigate whether genetic variants in IL-6 confer susceptibility to OSAS. The study population consisted of 151 patients with OSAS and 75 healthy controls from Southeast China. Five haplotype tagging single nucleotide polymorphisms (tSNPs) were selected across 21 kb of the IL-6 locus using Haploview software V4.1. The tSNPs were amplified by polymerase chain reaction (PCR) and genotyped by restriction enzyme digestion followed by gel electrophoresis. Linkage disequilibrium (LD) and haplotype reconstruction were carried out by means of a SHEsis program. No distribution difference of any of the five tSNPs between OSAS patients and controls was observed. However, in non-obese individuals (n=117), the minor allele G (rs1800796) decreased risk of OSAS compared with the major allele C [odds ratio (OR), 0.48; 95% confidence interval (CI), 0.26-0.86; p=0.014], and the haplotype TG (rs1880242, rs1800796) conferred a significantly decreased risk of OSAS than single allele G (rs1800796) (OR, 0.39; 95% CI, 0.20-0.74; p=0.003). Moreover, the severity of sleep disordered breathing (measured by apnea hypopnea index) increased linearly in carriers of the C variant of IL-6 -572G/C polymorphism (14.3+/-5.1, 22.0+/-3.6 and 34.8+/-3.5 for GG, CG and CC, respectively; p=0.012). To the best of our knowledge, this is the first study to suggest that genetic variants in IL-6 could modify OSAS susceptibility. SNP genotyping of IL-6 is a potential strategy for detecting the risk of breathing disordered diseases in non-obese individuals. PMID- 19288025 TI - Antiviral effect of arginine against herpes simplex virus type 1. AB - We investigated the effects of arginine on the multiplication of herpes simplex virus type 1 (HSV-1) and the potential of arginine as an antiherpetic agent. Arginine suppressed the growth of HSV-1 concentration-dependently. Inhibition of HSV-1 by arginine leveled off at 50-60 mM, although the higher concentration was not suitable as an antiviral agent due to cytotoxicity. 'Time of addition' experiments revealed that arginine was particularly effective when added within 6 h post-infection (h p.i.), suggesting that the reagent sensitive step is in the early stages of the infection. A one-step growth curve of HSV-1 in the presence of 30 mM arginine revealed that: i) the latent period was significantly extended, ii) the rate of formation of progeny infectious virus decreased and iii) the final yield of progeny virus decreased to 1%. The addition of arginine at 8 h p.i., after the completion of viral DNA replication in the virus multiplication, allowed the normal formation of progeny virus in the subsequent 4 h, confirming that arginine does not directly interfere with the formation of progeny infectious virus. In addition, arginine also inhibits several RNA viruses. PMID- 19288026 TI - Augmentation of the antimicrobial activities of guinea pig cathelicidin CAP11 derived peptides by amino acid substitutions. AB - Mammalian myeloid and epithelial cells express various peptide antibiotics (such as defensin and cathelicidin) that contribute to the innate host defense against invading microorganisms. Among these, guinea pig cathelicidin CAP11 (G1-I43) possesses potent antibacterial activities against Gram-positive and -negative bacteria, and also lipopolysaccharide-neutralizing activity. We previously revealed that the active region with antibacterial activity is localized at G1 to R18 of CAP11. In this study, to develop peptide derivatives with enhanced antimicrobial actions, we utilized the amphipathic 18-mer peptide (G1-R18) as a template. Anti-microbial activities of the peptides were assessed by alamarBlue assay (Escherichia coli, Staphylococcus aureus and Candida albicans) and colony formation assay (Porphyromonas gingivalis). Furthermore, the membrane permeabilization activities were determined by using E. coli ML-35p as a target. By substituting K5, T9, R10, R12, and G17 with five L residues, the hydrophobicity of the peptide (1-18m1) was increased, and by substituting G1, and Q14 with K and R residues, respectively, the hydrophilicity (positive charge) of the peptide (1-18m2) was enhanced. Among the peptides, 1-18m2 exhibits the most potent antimicrobial and membrane-permeabilizing activities against the microorganisms examined. Thus, the antimicrobial activities of the amphipathic CAP11-derived 18-mer peptide can be augmented by modifying its hydrophobicity and hydrophilicity (positive charge), and 1-18m2 is the most potent among the peptide derivatives with therapeutic potential for Gram-positive and -negative bacterial, and fungal infections. PMID- 19288027 TI - Evidence of a humoral response to a novel protein WARF4 embedded in the West Nile virus NS4B gene encoded by an alternative open reading frame. AB - West Nile virus (WNV) is a flavivirus that is maintained in a bird-mosquito transmission cycle. Humans, horses and other non-avian vertebrates are usually incidental hosts. However, WNV is a neurotropic virus, which requires an efficient humoral response for the control of a neuroinvasive infection. The WNV genome encodes three structural (capsid, premembrane/membrane and envelope) and seven non-structural proteins. Bioinformatic analysis performed on the WNV genomes detected a conserved alternative open reading frame restricted to the lineage I virus. To quickly verify the existence of this putative protein, entitled West Nile Alternative Reading Frame 4 (WARF4), we produced a prokaryotic recombinant source of WARF4 and verified its immunogenicity in vivo by analyzing 43 horse serum samples, of which 15 were positive for antibodies to WNV premembrane and envelope (prM-E) proteins. Specific antibodies to WARF4 were significantly detected in 5 out of the 15 serum samples testing positive for antibodies to prM-E WNV proteins. Our findings provide evidence of a significant antibody response to the WARF4 protein in the serum of the horse testing positive for antibodies to prM-E proteins, thus indicating that this antigen might be a potential tool for further characterization of the immune response of WNV infections in humans as well. PMID- 19288028 TI - Advanced glycation end products regulate extracellular matrix protein and protease expression by human glomerular mesangial cells. AB - Advanced glycation end products (AGEs) may play a role in the pathogenesis of diabetic nephropathy, by modulating extracellular matrix turnover. AGEs are known to activate specific membrane receptors, including the receptor for AGE (RAGE). In the present study, we analyzed the various receptors for AGEs expressed by human mesangial cells and we studied the effects of glycated albumin and of carboxymethyl lysine on matrix protein and remodelling enzyme synthesis. Membrane RAGE expression was confirmed by FACS analysis. Microarray methods, RT-PCR, and Northern blot analysis were used to detect and confirm specific gene induction. Zymographic analysis and ELISA were used to measure the induction of tPA and PAI 1. We show herein that cultured human mesangial cells express AGE receptor type 1, type 2 and type 3 and RAGE. AGEs (200 microg/ml) induced at least a 2-fold increase in mRNA for 10 genes involved in ECM remodelling, including tPA, PAI-1 and TIMP-3. The increase in tPA synthesis was confirmed by fibrin zymography. The stimulation of PAI-1 synthesis was confirmed by ELISA. AGEs increased PAI-1 mRNA through a signalling pathway involving reactive oxygen species, the MAP kinases ERK-1/ERK-2 and the nuclear transcription factor NF-kappaB, but not AP-1. Carboxymethyl lysine (CML, 5 microM), which is a RAGE ligand, also stimulated PAI 1 synthesis by mesangial cells. In addition, a blocking anti-RAGE antibody partially inhibited the AGE-stimulated gene expression and decreased the PAI-1 accumulation induced by AGEs and by CML. Inhibition of AGE receptors or neutralization of the protease inhibitors TIMP-3 and PAI-1 could represent an important new therapeutic strategy for diabetic nephropathy. PMID- 19288029 TI - Effect of MDR1 gene promoter methylation in patients with ulcerative colitis. AB - Altered MDR1 expression and/or function contribute to the pathogenesis of inflammatory bowel disease (IBD). DNA methylation was shown as an important mechanism in gene silencing. We investigated DNA methylation of the MDR1 gene in ulcerative colitis (UC) and its relation to MDR1 C3435T genotypes. Eighty-three UC patients were enrolled. Methylation of MDR1 promoter was determined by methylation specific polymerase (MSP) for rectal inflammatory mucosa from all patients and normal terminal ileum from 17 patients. Promoter methylation of MDR1 gene was also quantified by digital densitographic analysis following MSP. MDR1 methylation was detected in 51 (61.4%) out of 83 patients in rectal inflammatory mucosa. Mean methylation level of MDR1 gene in rectal inflammatory mucosa was significantly higher than in normal terminal ileum (p=0.021). MDR1 methylation occurred more frequently in total colitis, and total+left side colitis, compared to rectal colitis (p=0.001, 0.013, respectively). Higher methylation levels were also associated with chronic continuous type (p=0.034) and earlier onset of disease (p=0.038). The 3435 CC+CT genotype of MDR1 was associated with more than 6-fold increased risk of MDR1 methylation, especially in UC patients with 9 years and shorter duration. Both frequency and level of MDR1 methylation were higher in UC onset at younger or in middle age with the same genotype. MDR1 methylation frequently occurred in inflammatory rectal mucosa from UC patients and was influenced by MDR1 C3435T polymorphism, especially in patients with shorter duration and younger onset. PMID- 19288030 TI - Association of genetic variants with chronic kidney disease in Japanese individuals with type 2 diabetes mellitus. AB - Although diabetes mellitus has been recognized as a risk factor for chronic kidney disease (CKD), genetic factors for predisposition to CKD in individuals with diabetes mellitus remain elucidated. The purpose of the present study was to identify genetic variants that confer susceptibility to CKD among individuals with type 2 diabetes mellitus. The study population comprised 1742 Japanese individuals, including 636 subjects with CKD [estimated glomerular filtration rate (eGFR)<60 ml/min/1.73 m2] and 1106 controls (eGFR>or=60 ml/min/1.73 m2). The genotypes for 24 polymorphisms of 22 candidate genes were determined. An initial screen of allele frequencies by the Chi-square test revealed that four polymorphisms were significantly (false discovery rate<0.05) associated with the prevalence of CKD in individuals with type 2 diabetes mellitus. Subsequent multivariable logistic regression analysis with adjustment for covariates as well as a stepwise forward selection procedure revealed that the 8733T-->C polymorphism of ALOX5AP (rs3803278), the C-->T (Ser532Leu) polymorphism of IRAK1 (rs1059703), and the 2445G-->A (Ala54Thr) polymorphism of FABP2 (rs1799883) were significantly (P<0.05) associated with the prevalence of CKD. Our results suggest that ALOX5AP, IRAK1, and FABP2 are susceptibility loci for CKD among Japanese individuals with type 2 diabetes mellitus. PMID- 19288031 TI - Cerebellin and des-cerebellin exert ACTH-like effects on corticosterone secretion and the intracellular signaling pathway gene expression in cultured rat adrenocortical cells--DNA microarray and QPCR studies. AB - Precerebellins (Cbln) belong to the C1q/TNF superfamily of secreted proteins which have diverse functions. They are abundantly expressed in the cerebellum, however, three of them are also expressed in the rat adrenal gland. All members of the Cbln family form homomeric and heteromeric complexes with each other in vitro and it was suggested that such complexes play a crucial role in normal development of the cerebellum. The aim of our study was to investigate whether Cbln1-derived peptides, cerebellin (CER) and des-Ser1-cerebellin (desCER) are involved in regulating biological functions of rat adrenocortical cells. In the primary culture of rat adrenocortical cells, 24 h exposure to CER or desCER notably stimulated corticosterone output and inhibited proliferative activity and similar effects were evoked by ACTH. To study gene transcript regulation by CER, desCER and ACTH, we applied Oligo GEArray DNA Microarray: Rat Signal Transduction Pathway Finder. In relation to the control culture, 13 of the 113 transcripts present on the array were differentially expressed. These transcripts were either up- or down-regulated by ACTH and/or CER or desCER treatment. Validation of DNA Microarray data by QPCR revealed that only 5 of 13 genes studied were differentially expressed. Of those genes, Fos and Icam1 were up-regulated and Egr1 was down-regulated by ACTH, CER and desCER. The remaining two genes, Fasn (insulin signaling pathway) and Hspb1 (HSP27) (stress signaling pathway), were regulated only by CER and desCER, but not by ACTH. Thus, both CER and desCER have effects similar to and different from corticotrophin on the intracellular signaling pathway gene expression in cultured rat adrenocortical cells. PMID- 19288032 TI - Fructus Xanthii extract protects against cytokine-induced damage in pancreatic beta-cells through suppression of NF-kappaB activation. AB - Cytokines released by infiltrating inflammatory cells around the pancreatic islets are involved in the pathogenesis of type 1 diabetes. Interleukin (IL) 1beta and interferon (IFN)-gamma are the primary cytokines responsible for stimulation of inducible nitric oxide synthase (iNOS) expression and nitric oxide overproduction, which leads to beta-cell damage. In addition, nuclear factor kappaB (NF-kappaB) plays a crucial role in the activation of this pathway. Therefore, suppression of the cytokine-NF-kappaB pathway is considered an effective therapeutic strategy for preventing inflammatory reactions in pancreatic beta-cells. In this study, the effects of Fructus Xanthii extract (FXE) on IL-1beta and IFN-gamma-induced beta-cell damage were examined. Treatment of RINm5F cells with IL-1beta and IFN-gamma reduced cell viability, however, FXE completely protected cells from IL-1beta and IFN-gamma-mediated reduction in viability in a concentration-dependent manner. In addition, incubation with FXE resulted in a significant suppression of IL-1beta and IFN-gamma-induced nitric oxide (NO) production, which correlated with the reduced levels of the inducible form of iNOS mRNA and protein observed. The IL-1beta and IFN-gamma-stimulated RIN cells showed increases in NF-kappaB binding activity and p50 subunit levels in the nucleus, as well as increased IkappaBalpha degradation in cytosol when compared to unstimulated cells, which indicates that the mechanism by which FXE inhibited the iNOS gene involves inhibition of NF-kappaB activation. Furthermore, a protective effect of FXE was demonstrated by reduction in NO generation and iNOS expression, as well as the normal insulin secreting responses to glucose observed in IL-1beta and IFN-gamma-treated islets. PMID- 19288033 TI - Trans-Zeatin inhibits UVB-induced matrix metalloproteinase-1 expression via MAP kinase signaling in human skin fibroblasts. AB - Ultraviolet (UV) irradiation induces the expression of matrix metalloproteinases (MMPs), disturbing the metabolism of extracellular matrix (ECM), and causes the characteristic changes of photoaging in skin. Inhibition of induction of MMPs is suggested to alleviate photoaging induced by UV irradiation. Zeatin, purified from Zea mays, is a member of the cytokinin group of plant growth factors, the activity of which is attributed to its more stable trans form. In this study, we investigated the effect of trans-Zeatin on UVB-induced matrix metalloproteinase-1 (MMP-1) expression in cultured human skin fibroblasts (HSFs) and studied the mechanisms of its actions. We found that pretreatment with trans-Zeatin significantly inhibits UVB-induced MMP-1 expression and c-Jun activation in a dose-dependent manner. We also observed that trans-Zeatin inhibits UVB-induced phosphorylation of ERK, JNK and p38 MAP kinases (MAPKs) dose-dependently. As expected, PD98059, an ERK inhibitor, SP600125, a JNK inhibitor and SB203580, a p38 MAPK inhibitor effectively inhibit UVB-induced phosphorylation of ERK, JNK and p38 MAPKs, respectively. Moreover, the inhibitory mechanism of trans-Zeatin was further demonstrated in MMP-1 secretion using MAPK-specific inhibitors. PD98059, SP600125 and SB203580 suppressed UVB-induced MMP-1 secretion, which is consistent with the above results. Collectively, our results suggest that trans Zeatin inhibits UVB-induced MMP-1 expression, which may be mediated by inhibition of ERK, JNK and p38 MAPKs signaling pathways in HSFs. Trans-Zeatin is a potential agent for the management of skin photoaging. PMID- 19288034 TI - Therapeutic target validation of protein kinase C(PKC)-zeta for asthma using a mouse model. AB - Protein kinase C (PKC) is a complex family consisting of many types of isoenzymes, of which PKC-zeta, an atypical isoform, has been reportedly implicated in the regulation of apoptosis and NF-kappaB, as well as control of T dependent responses. Based on the recent report that PKC-zeta controls TH2 response, the current study was aimed to evaluate PKC-zeta as a potential therapeutic target for asthma using a mouse model. Mouse allergic asthma was induced by repeated sensitization followed by intranasal challenge with OVA and PKC-zeta pseudosubstrate inhibitor (PPI) was intratracheally instilled before each OVA challenge. Airway hyperreactivity (AHR) was measured by beta-methacoline induced airflow obstruction. Cellular and cytokine profile in bronchoalveolar lavage fluid (BALF) and level of serum IgE as well as cytokine production by draining lymph node cells were compared. AHR and numbers of eosinophils in BALF were significantly lowered by PPI, indicating that blocking of PKC-zeta activation alleviates asthmatic manifestations. Additionally, PPI instillation decreased IL-5 and IL-13 levels in BALF to approximately 20% of controls, but not IFN-gamma level. Instillation of PPI also caused a marked fall in the level of TNF-alpha, another NF-kappaB-dependent, proinflammatory cytokine. Serum OVA specific IgE level and ex vivo IL-4, IL-5 and IL-13, but not IFN-gamma, production by peribronchial lymph node cells was also considerably lower in PPI treated mice. In conclusion, blockade of PKC-zeta signals by intratracheal instillation of PPI alleviates allergen-specific TH2 response as well as asthmatic manifestations and hence PKC-zeta is a promising target for treatment of asthma. PMID- 19288035 TI - A Spanish version of the Family Assessment Device. AB - INTRODUCTION: This paper presents the results of a study on the psychometric properties of an authorized Spanish version of the McMaster Family Assessment Device, a self-report measure of family functioning. MATERIALS AND METHODS: The study sample comprised 60 psychiatric patients and their family member and 60 controls, without mental health problems, and their family member. RESULTS: Compared to other studies, all subscales displayed adequate temporal stability and acceptable reliability. While the instrument discriminated well between the two groups of families on all subscales, the results nevertheless indicated limitations in the inter-item discriminant capacity of the "Roles" subscale. Factor analysis resulted in a three-factor model that does not coincide with the established structure of this instrument. CONCLUSION: Proposals to improve and adapt questionnaire are discussed with a view to make it applicable to cultures other than the one it was developed. Theoretical models relating to psychosocial aspects such as family functioning, albeit compatible in some areas, should be viewed with caution in cultures different to that in which the model originates. PMID- 19288036 TI - Cumulative adversity and depressive symptoms among older adults in Israel: the differential roles of self-oriented versus other-oriented events of potential trauma. AB - BACKGROUND: The study examined the association between cumulative adversity and current depressive symptoms in a national sample of Israelis aged 50+. Referring to cumulative adversity as exposure to potentially traumatic events along life, the study distinguished between events primarily inflicted upon the self (self oriented adversity) versus upon another person (other-oriented adversity). METHOD: Data were drawn from the Israeli component of the Survey of Health, Aging and Retirement in Europe (SHARE). During 2005-2006, 1710 Jews and Arabs completed an inventory of potentially traumatic events and two measures of depressive symptoms: the European Depression scale (Euro-D) and the Adapted Center for Epidemiological Studies-Depression scale (ACES-D). The Euro-D is more detailed in querying cognitions and motivations while the ACES-D is more detailed in querying feelings and social alienation. RESULTS: In line with the hypothesis, self oriented adversity had a positive association with depressive symptoms whereas other-oriented adversity had either no association or an inverse association with depressive symptoms. Sociodemographic characteristics and perceived health were controlled in the multivariate regressions. CONCLUSIONS: The differential association of self- versus other-oriented adversity with depressive symptoms may be explained in terms of social commitments that are inherent in other-oriented adversity and incompatible with depressive symptoms. The study points to the variations in the symptom compositions represented by the Euro-D and ACES-D, with the latter better capturing the difference between self- and other-oriented adversities. PMID- 19288037 TI - Assessment of the environmental quality of French continental Mediterranean lagoons with oyster embryo bioassay. AB - In order to better understand environmental disturbances in the French coastal Mediterranean lagoons, we used an ecotoxicological approach based on the measurement of the toxicity of the sediments using oyster embryo bioassay that provides a basis for assessing the effects on the fauna of contaminants adsorbed on the sedimentary particles. The study covers all of the main lagoons of the French Mediterranean coasts of Languedoc Roussillon, Camargue, and Provence (Berre and Bolmon lagoons), where 188 stations were sampled. The toxicity tests provide evidence of variable levels of toxicity in sediments. Contaminated lagoons such as La peyrade, Le canet, and Ingrill and locally affected lagoons such as Bages-Sigean, Vaccares, Bolmon, and Berre have sampling stations with 100% of larval abnormalities during 24-h development. In all of the lagoons, the toxicity was mainly located close to local harbors and rivers. Salses Leucate (Languedoc roussillon) lagoon was found very clean, with no important toxicity. The results are discussed in terms of environmental disturbances of the coastal lagoons and with regard to the long-term monitoring of the impact of contaminants on the coastal environment. PMID- 19288038 TI - Characteristics of esophagorespiratory fistulas resulting from esophageal cancers: a single-center study on 243 cases in a 20-year period. AB - BACKGROUND: Esophagorespiratory fistulas are serious complications of esophageal tumors. The main goal of this study was to reveal their characteristics. METHODS: In the period between 1984 and 2004, 243 fistulas were diagnosed among the 1993 patients with esophageal cancer. Comparing the characteristic data of patients with and without fistula (demographics, symptoms, duration time of symptoms, morphologic features, histology, metastases, staging), multivariance analysis showed significant differences (P < 0.05). Patients with fistula were divided into two additional groups (I, n = 82; II, n = 161 cases) with cluster analysis. The difference between these two groups was also significant. RESULTS: It was possible to perform a palliative endoscopic intubation in 176 cases, while 62 patients could be treated only supportively. The average survival was 3.4 and 1.2 months, respectively. Fistula was a late complication of tumor in two thirds (66.3%) of the cases, while in 33.7% it was diagnosed in younger patients at the early stage of the disease, with a more aggressive, less differentiated histology. In these patients the weight loss, the grade of dysphagia, and the size of the tumor were smaller, the possibilities of treatment were fewer, and survival time was shorter (2.1 vs. 3.1 months). CONCLUSIONS: These tumors seem to be specific forms of esophageal cancers. For a better quality of life and longer survival time for these patients, there should be earlier diagnosis and endoscopic intubation as the best palliative treatment should be performed. PMID- 19288040 TI - Greater trochanter displacements after Charnley's trochanterotomy, during passive hip movements: a real-time 3D cadaveric approach. AB - Non-unions of greater trochanter (GT) fractures or osteotomies are the source of frequent complications. Two muscles are involved in the genesis of such non unions, the gluteus medius and the gluteus minimus. Literature says that, during hip flexion, their terminal insertions on the faces of the GT generate an anterior translation of the GT responsible for contact loosening and healing defect. The main goal of this paper was to assess this hypothesis into the 3D space and to quantify the 3D displacements (three translations and three rotations) of the GT after trochanterotomy during passive motions of the hip joint (flexion, abduction-adduction, internal and external rotation). Therefore, we used 13 fresh cadaveric hips, on which we registered the GT displacements thanks to a 3D infrared localizer. An ultra low weight dynamic reference based has been developed to track the motion of the GT without any interference. The results have shown rotatory displacements of the GT around the three axes. This produces a posterior and lateral opening of the osteotomy site associated with a posterior rotatory shear of the GT. Above 45 degrees of hip flexion these three rotations combine together to exert shear on the osteotomy site which is harmful to the osteosynthesis material and the consolidation potential and may explain implant loosening. PMID- 19288039 TI - Efficacy of chlorhexidine gluconate during surgery for hydatid cyst. AB - BACKGROUND: Dissemination of protoscolices-rich fluid during surgery for hydatid cyst disease is a major cause of recurrence. Instillation of a scolicidal agent into a hepatic hydatid cyst before opening is the most commonly employed measure to prevent this serious complication. In this clinical study, the efficacy of 0.04% chlorhexidine gluconate (Chx-Glu) during hydatid cyst surgery was tested, and early-term results were evaluated. METHODS: A total of 30 consecutive patients with 45 liver cysts were studied. Only type I and type II cysts were included. Cysts were punctured, and fluid was aspirated. The viability of protoscolices in this fluid was determined. Chx-Glu 0.04% was instilled into the cyst cavity. After 5 minutes of exposure, the cyst fluid was reaspirated and evaluated for the viability of protoscolices. Imprints of the germinative membranes were determined for protoscolices viability. RESULTS: All protoscolices in 45 cysts evaluated were killed by 5 minutes of exposure to 0.04% Chx-Glu. The whole of the germinative membrane imprints contained dead protoscolices. Chx-Glu 0.04% did not cause any adverse effect on biliary tracts in communication with cysts. There was no recurrence during the 2-year follow-up period. There were also no mortality and no cavity-related complications. CONCLUSIONS: This study shows that intracystic injection of 0.04% Chx-Glu is an effective measure against the dissemination of viable protoscolices. In addition, Chx-Glu is the most convenient scolicidal agent as per the criteria defined by the World Health Organization. Therefore, it can be used safely during hydatid cyst surgery. However, there remains the need to perform advanced comparative clinical studies on the efficacy of Chx-Glu and other scolicidal agents. PMID- 19288041 TI - Ex vivo and in vivo coronary ostial locations in humans. AB - PURPOSE: Knowledge of the normal in vivo distribution and variation of coronary ostial locations is essential in the planning of various interventional and surgical procedures. However, all studies to date have reported the distribution of coronary ostia locations only in cadaver hearts. In this study, we sought to assess the distribution of coronary ostial locations in patients using cardiac dual-source computed tomography (CT) and compare these values to those of human cadaveric specimens. METHODS: Measurements of the coronary ostia location were performed in 150 patients undergoing dual-source CT and in 75 cadavers using open measurement techniques. All 150 patients had a normal aortic valve function and no previous cardiac intervention or surgery. The location of the right and left coronary origin in relation to the aortic annulus and the height of the sinus of Valsalva were measured. RESULTS: Mean ostial locations at CT were 17.0 (+/-3.6) mm and 15.3 (+/-3.1) mm for the right and left coronary ostia, with large variations of both sides (right: 10.4-28.5 mm; left: 9.8-29.3 mm). In cadavers, mean locations were 14.9 (+/-4.3) mm [5-24 mm] for right and 16.0 (+/-3.6) mm [9 24 mm] for left coronary ostia. Comparison of CT and cadaver data showed statistically significant differences for right (P < 0.0001) but not left (P = 0.1675) coronary ostia. CONCLUSIONS: This study provides data of normal coronary ostial origins and demonstrates significant differences between in vivo and ex vivo measurements regarding the right coronary ostium. The observed large variations of coronary ostia origins emphasize the importance of considering such anatomic variations in the development of treatments. PMID- 19288042 TI - Prevalence and definition of drooling in Parkinson's disease: a systematic review. AB - Drooling (saliva loss) is a frequently reported symptom in patients with Parkinson's disease (PD), but an accurate estimate of the prevalence of drooling is lacking. The aim of this study was to systematically review the prevalence of drooling in published research papers. A systematic PubMed and CINAHL search was done, including studies published until January 2009. Eight studies were found, presenting prevalence rates of drooling based on responses of PD patients to questionnaires. The statistical heterogeneity was highly significant (P < 0.0001), with prevalence rates ranging from 32 to 74%. The pooled prevalence estimate with random effect analysis was of 56% (95% CI 44-67) for PD patients and 14% (95% CI 3-25) for healthy controls; the pooled relative risk (RR) with random effect analysis was 5.5 (95% CI 2.1-14.4). All studies reported data of community dwelling idiopathic PD patients, with a mean age around 65 years and mild PD in 50-60% of the cases. Heterogeneity was mainly caused by differences in definition or frequency of drooling. The highest prevalence rates included nocturnal drooling where others noted only diurnal drooling. Analysis of the data of two studies showed that drooling is reported frequently by 22-26% of the patients. Prevalence rates were lower in milder PD patients. The summarized findings demonstrate that drooling can be present in half of all PD patients. In about a quarter of PD patients, drooling appears to be a frequently occurring problem. We recommend to report drooling in future studies with more detailed consideration of severity, frequency and nocturnal versus diurnal complaints. PMID- 19288043 TI - Bilateral brachial neuropathy and acute aortic dissection due to giant cell arteritis. PMID- 19288044 TI - Headaches in antiquity and during the early scientific era. AB - This paper presents the evolution of ideas on headache symptoms from antiquity through the 19th century. A thorough study of texts, medical books and reports along with a review of the available literature in PubMed was undertaken: observations on headaches date back nearly 4,000 years to the ritual texts of Mesopotamia. Nicolaes Tulp, Thomas Willis and Gerhard van Swieten also made important contributions on various forms of headaches in the 17th and 18th centuries. Edward Liveing and William Gowers made the major contributions to the field in the late 19th century. Overall, observations on headaches span a timeline of nearly 9,000 years. The work of the physicians during the 18th and 19th century, however, set the basis for scientific research. PMID- 19288045 TI - Quality of life and neurobehavioral changes in survivors of malignant middle cerebral artery infarction. AB - Malignant middle cerebral artery (MMCA) infarction is associated with a mortality rate of 80% under conservative treatment. Decompressive hemicraniectomy (DH) reduces mortality and improves the functional outcome of surviving patients. The purpose of this study was to examine quality of life (QoL) and neurobehavioral deficits in patients with space-occupying infarctions of the right- or left-sided hemisphere at 6 months after stroke. The Sickness Impact Profile (SIP) was used to assess QoL in 19 out of 29 consecutive patients that underwent DH after a malignant MCA infarction (14 on the right and 5 on the left hemisphere). Behavioral changes were evaluated with the Frontal Behavioral Inventory and the Beck Depression Inventory. Patients and relatives were also asked if, knowing the present outcome, they would agree again, in retrospect, to a DH. Barthel Index >60 was seen in 37% of our patients. Functional outcome was related to age. We found a higher reduction in the SIP's physical domain than in the psychosocial domain. Depressive symptoms were present in 50% of the patients. We didn't find significant differences in QoL or functional outcome between patients with right or left-sided infarctions. The most frequent neurobehavioral symptoms were decreased speech output, apathy, reduced spontaneity and irritability. Most patients and their relatives would again give consent to hemicraniectomy. The results show that younger patients had a significantly better outcome. QoL seems to be acceptable in both left- and right-sided infarctions, and retrospective agreement to hemicraniectomy is high in both patients and their relatives. PMID- 19288046 TI - A preliminary study of the frequency of anti-basal ganglia antibodies and streptococcal infection in attention deficit/hyperactivity disorder. AB - Attention deficit/hyperactivity disorder (ADHD) is often present in patients with post-streptococcal neuropsychiatric disorders such as Sydenham's chorea and PANDAS, in which anti-basal ganglia antibodies (ABGA) have been frequently found. Our study investigates the hypothesis that pharyngeal group A beta-hemolytic streptococcus (GABHS) infections and serum ABGA are more frequent in children with ADHD non-comorbid (nc-ADHD) with obsessive-compulsive disorder or tics than in controls. We compared 22 children with nc-ADHD (DSM-IV-TR) and 22 healthy controls matched by age, gender and season of sample collection, for the frequency of recent GABHS infection and the presence of ABGA. Eleven out of 22 children (51%) with nc-ADHD showed evidence of GABHS infection compared to three out of 22 (14%) controls (P = 0.007). We found positive ABGA in one ADHD subject (4%) and in one control (4%). This preliminary study indicates that frequency of ABGA in children with nc-ADHD does not differ from that in matched controls, despite the fact that our ADHD patients had had more recent GABHS infections than the controls. This suggests that ABGA do not have a role in the pathogenesis of nc-ADHD. PMID- 19288047 TI - The safety and efficacy of clopidogrel versus ticlopidine in Japanese stroke patients: combined results of two Phase III, multicenter, randomized clinical trials. AB - Two Phase III studies comparing the safety and efficacy of clopidogrel with ticlopidine as antiplatelet agents for the secondary prevention of vascular events in patients with prior stroke were performed in Japan. Both studies were randomized, double-blind, double-dummy comparative trials with the primary objective of comparing the clinical safety of treatment with either clopidogrel or ticlopidine for up to 12 months. The secondary objective was to assess the incidence of a combined efficacy endpoint of cerebral infarction, myocardial infarction, and vascular death. Patients with prior stroke were recruited during July 1996-February 1998 and September 2001-November 2003 at centers across Japan. The results of the two studies were combined in this analysis. There were 1,869 patients in the safety population (clopidogrel, 941; ticlopidine, 928). Significantly, fewer patients experienced a safety event in the clopidogrel group than in the ticlopidine group (p < 0.001; hazard ratio, 0.610; 95% confidence interval 0.529, 0.703). Almost twice as many patients in the ticlopidine group (25.6%) experienced hepatic dysfunction than in the clopidogrel group (13.4%). There were 1,862 patients evaluable for efficacy (clopidogrel, 939; ticlopidine, 923). There was no significant difference in the incidence of the combined efficacy endpoint between clopidogrel (2.6% of patients) and ticlopidine (2.5%). Clopidogrel was better tolerated than ticlopidine. There was no difference in the efficacy of the two agents with regard to the secondary prevention of vascular events in patients with prior stroke. This was the first combined analysis of direct comparison of clopidogrel with ticlopidine in the clinical setting. PMID- 19288048 TI - Small-cell carcinoma in the common bile duct treated with multidisciplinary management. AB - Small-cell carcinoma occasionally occurs in the gastrointestinal tract, with rare reports of it in the biliary system. When it does occur in the biliary system, it is mainly within the gallbladder, and those arising in the extrahepatic bile duct are extremely rare with only seven reported cases to date. A 62-year-old man with this unusual condition is documented. The patient was preoperatively diagnosed as having small-cell carcinoma on the basis of a biopsy specimen and was treated with multidisciplinary management, which consisted of preoperative chemotherapy, a curative resection, adjuvant chemotherapy, and radiation therapy. The patient died 23 months after the initial diagnosis (20 months after surgery). To the best of our knowledge, the survival time of this case was the longest among reported cases, and multidisciplinary management might contribute to a prolonged survival. PMID- 19288049 TI - Hepatocellular adenoma in a male with familial adenomatous polyposis coli. AB - Hepatocellular adenoma (HA) is a benign liver tumor most frequently occurring in young women using oral contraceptives. We report a rare case of HA in a 27-year old male patient with familial adenomatous polyposis (FAP). The patient underwent a total colectomy and ileo-rectal anastomosis for FAP in 2003. A preoperative computed tomography scan of the abdomen disclosed a tumor in the left-lobe of the liver, 5.8 cm in diameter. Pathologic examination of a needle biopsy disclosed HA, but he had never used anabolic steroids or other known inducers of HA. The size of the liver mass gradually increased to 8.5 cm during a follow-up period of 38 months, and a left hepatectomy was performed in 2006. Pathology of the resected specimen confirmed the diagnosis of HA. Although FAP is known to be complicated with neoplasia in various extracolonic organs, only five reported cases of HA have developed in patients with FAP, including this case. This is the first report of HA to develop in a male FAP patient. PMID- 19288050 TI - Identification of the CLCN7 gene mutations in two Chinese families with autosomal dominant osteopetrosis (type II). AB - Here we report the identification of two different mutations in chloride channel 7 gene in two unrelated patients with autosomal dominant osteopetrosis type II. We determined that one patient (a 32-year-old woman) carried a heterozygous gene for a R767W mutation in exon 24, and another patient (a 17-year-old boy) carried a heterozygous gene for a novel frameshift mutation (Glu798FS) in exon 25. Recent studies have reported loss-of-function mutations in the chloride channel 7 (CLCN7) gene as a cause of autosomal dominant osteopetrosis type II (ADO-II). The identification of gene mutations in Chinese with ADO has not been reported previously. In this study, we identified mutations of the CLCN7 gene in two unrelated Chinese families with ADO-II. Two probands with ADO-II were diagnosed based on their bone characteristics on X-rays and their laboratory results. All 25 exons of the CLCN7 gene, including the exon-intron boundaries, were sequenced. We found in family 1 that the proband (a 32-year-old woman) was heterozygous for a CLCN7 mutation. The nonsynonymous mutation consisted of a heterozygous C/T transition at codon 2327 in exon 24, which resulted in an arginine (CGG)-to tryptophan (TGG) substitution at position 767 (R767W). The same heterozygous mutation (C/T) was determined in her father and son, who were asymptomatic with normal skeleton radiography. In family 2, we found that the proband (a 17-year old boy) carried a novel frameshift mutation (Glu798FS) resulting from a G insertion between codon 60 and codon 61 in exon 25. The heterozygous -/G insertion is predicted to elongate the peptide of CLCN7 by 120 amino acids after position 797 amino acids. Similarly, some individuals of this family carried the same heterozygous mutation, but they are all asymptomatic. Furthermore, the R767W and Glu798FS mutations were not found in 100 unrelated controls. Our present findings suggest that the novel Glu798FS mutation in exon 25 and R767W in exon 24 in the CLCN7 gene were responsible for ADO-II in these Chinese patients. PMID- 19288051 TI - The complete nucleotide sequence of the RNA2 of the crinivirus tomato infectious chlorosis virus: isolates from North America and Europe are essentially identical. AB - The complete nucleotide sequences of the RNA2 of two isolates of Tomato infectious chlorosis virus (TICV, genus Crinivirus, family Closteroviridae) from the United States and Spain, respectively, were determined. The sequences of both isolates were found to be nearly identical. TICV RNA2 consisted of 7,914 nucleotides in both isolates and contains eight open reading frames that encompass the Closteroviridae hallmark gene array represented by a heat shock protein 70 family homologue, a protein of 59 kDa, the major coat protein, and a divergent copy of the coat protein. Phylogenetic analysis suggested that TICV is most similar to Lettuce infectious yellows virus (LIYV), the type species of the genus Crinivirus. PMID- 19288052 TI - A comparison of the pathogenicity of avian and swine H5N1 influenza viruses in Indonesia. AB - Highly pathogenic avian influenza H5N1 viruses are circulating in many countries. We recently discovered that these viruses have been transmitted to pigs on multiple occasions in Indonesia. To investigate whether avian H5N1 influenza viruses adapted to mammals through their introduction into pigs, we examined the growth of avian and swine isolates in cell culture and compared their pathogenicity in mice. We found that swine isolates were less virulent to mice than avian isolates, suggesting that the viruses became attenuated during their replication in pigs. Continuous surveillance of H5N1 viruses among pigs is clearly warranted. PMID- 19288053 TI - Molecular characterization of a foot-and-mouth disease virus containing a 57 nucleotide insertion in the 3'untranslated region. AB - A foot-and-mouth disease virus containing a 57-nucleotide (nt) insertion in the 3'untranslated region (3'UTR) was generated by transposon (tn)-mediated mutagenesis. Characterization of the mutant virus (A24-3'UTR8110) revealed no significant differences in virus growth, translation efficiency or virulence in cattle compared to the A24 wild-type virus. RNA modeling showed that the structures predicted in the 3'UTR were not affected by the tn insertion. These results revealed that the 3'UTR can tolerate foreign sequences that do not disrupt essential signals required for virus replication. PMID- 19288054 TI - Acid-etch interval and shear bond strength of Er,Cr:YSGG laser-prepared enamel and dentin. AB - The formation of a superficial layer of tiny flakes has been observed on teeth prepared by Erbium lasers. It has been suggested that removing this layer (mechanically or chemically) may increase the bond strength of the resin composite. The purpose of this study is to evaluate the effect of various etching times on bond strength of resin composite to enamel and dentin prepared by Er,Cr:YSGG laser. Sixty previously flattened human molars were irradiated for 10 s by an Er,Cr:YSGG laser. Enamel (E) specimens were etched with 37% H(3)PO(4) for 20, 40 or 60 s and dentin (D) specimens were etched for 15 or 30 s. All specimens were prepared for a standard shear bond strength (SBS) test (1 mm/min). Data were analyzed [ANOVA, Tukey post-hoc, a < 0.05)] and the failure mode was studied under SEM. Mean SBS+/-sd (MPa) for each group was 16.97 +/- 7.77 (E20s), 21.34 +/ 3.55 (E40s), 14.08 +/- 4.77 (E60s), 13.62 +/- 7.28 (D15s) and 13.15 +/- 6.25 (D30s). SBS for E40s was significantly higher than E60s (p = 0.023). No difference was noted between the dentin groups. SEM evaluation showed predominantly cohesive failure. Within the limits of this study, etching time significantly influenced the SBS of composite resin to laser-prepared enamel. SEM showed subsurface cracks, fissures, and deformities leading to predominantly cohesive failure in both enamel and dentin. PMID- 19288055 TI - Sneddon's syndrome presenting with severe disabling bilateral headache. AB - Sneddon's syndrome is a rare vascular disease affecting mainly skin and brain arterioles leading to their occlusion due to excessive endothelial proliferation. The two main features of this syndrome are livedo reticularis and lacunar subcortical infarcts. Here, we describe the case of a 64-year-old woman presenting with a 4-year history of a throbbing, bilateral, parieto-occipital headache associated with facial pain, but without any other accompanying symptom. The pain, initially misdiagnosed as atypical trigeminal neuralgia, worsened up to chronic daily and such severely disabling headache that she was constrained to bed. She presented with reduced cognitive functions, diffuse and severe livedo reticularis, severe myalgias and mild stiffness. All diagnostic test for different diseases were performed and other diseases excluded except for Sneddon's syndrome. Her symptoms were reduced firstly using acetylsalicylic acid, then ticlopidine 250 mg bid was begun and then Pentoxyphillin, resulting in a significant improvement of symptoms with the disappearance of headache. Her worsening in the first year was characterized by obsessive-compulsive behaviours, body-image misperceptions and panic attacks, improved for a period using olanzapine. Considering this case, we remark the importance of using headache classification to avoid diagnostic errors, secondly, we describe an atypical manifestation of Sneddon's syndrome and therapeutic efficacy of using ticlopidine and pentoxyphillin. PMID- 19288056 TI - Strategies for the analysis of osteitic bone defects at the diaphysis of long bones. AB - Septic diseases of the bone and the immediate surrounding soft tissue, i.e., osteitis, belong to the most alarming findings in recent traumatology and orthopedic surgery. The paramount goal of this therapy is to preserve the stable weight-bearing bones while maintaining a correct axis and proper working muscles and joints, in order to avoid permanent disability in the patient. "State-of-the art" therapy of osteitis/osteomyelitis therapy has two priorities: eradication of the infection and reconstruction of bone and soft tissue. Surgical treatment of the affected bone segments and soft tissue, followed by reconstructive methods, continues to be the main basic therapy. It is often extremely difficult to decide whether the affected bone segment has to be resected, or whether bone continuity can be preserved. The following paper provides strategies and guidance to help guide decisions in this complex and challenging area. PMID- 19288057 TI - Digoxin and ouabain increase the synthesis of cholesterol in human liver cells. AB - Digoxin and ouabain are steroid drugs that inhibit the Na(+)/K(+)-ATPase, and are widely used in the treatment of heart diseases. They may also have additional effects, such as on metabolism of steroid hormones, although until now no evidence has been provided about the effects of these cardioactive glycosides on the synthesis of cholesterol. Here we report that digoxin and ouabain increased the synthesis of cholesterol in human liver HepG2 cells, enhancing the activity and the expression of the 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), the rate-limiting enzyme of the cholesterol synthesis. This effect was mediated by the binding of the sterol regulatory element binding protein-2 (SREBP 2) to the HMGCR promoter, and was lost in cells silenced for SREBP-2 or loaded with increasing amounts of cholesterol. Digoxin and ouabain competed with cholesterol for binding to the SREBP-cleavage-activating protein, and are critical regulators of cholesterol synthesis in human liver cells. PMID- 19288059 TI - [Palliative medicine. Fifth pillar of anaesthesia departments]. AB - Palliative medicine has progressed during recent years to an independent medical faculty within the German health system. Despite this development palliative care systems for out-of-hospital and in-hospital palliative care are still insufficient in Germany so that the development of necessary resources must be considered as not yet completed. To support the further national development palliative medicine can be temporarily or permanently coupled to existing departments, which can be advantageous for all concerned and last but not least be profitable to patients and their relatives. Possibilities for participation of anaesthesiologists in this area of medical care are discussed in the study reported here. Anaesthesiologists have always historically been represented in palliative medical departments, e.g. as pain specialists. In the following investigation the special possibilities of anaesthesia departments for supporting the education and development of in-hospital and out-of hospital palliative medical care departments are reported. Previous experience of co-operation between these two departments is well established. Departments of palliative medicine depend on a well working interdisciplinary co-operation between different medical disciplines (e.g. anaesthesiology, radiotherapy, surgery and oncology) and several medical professions (e.g. physicians, nurses, psychologists). The aim of palliative care therapy is to be responsible for the best possible therapy for cancer patients and to give support to their care giving relatives. Due to the increasing establishment of palliative care procedures in Germany, departments of anaesthesiology should actively take part in the further development. Part of the responsibility of most anaesthesia departments is to practice pain management and critical care medicine, which are reasons why anaesthesiologists are predestined to be part of the system for palliative care patients and their relatives. Anaesthesia departments can be responsible for the organization of in-hospital and out-of-hospital palliative medicine and palliative care. The integration of anaesthesiological expertise into palliative medicine departments and vice versa can be a great opportunity for both medical departments and therefore represents a worthwhile engagement. PMID- 19288060 TI - [Role of the anaesthesiologist in an interdisciplinary field]. PMID- 19288062 TI - Regulation of angiogenesis by oxygen sensing mechanisms. AB - The choices for blood vessels to undergo angiogenesis or stay quiescent are mostly determined by the status of tissue oxygenation. A major link between tissue hypoxia and active angiogenesis is the accumulation of hypoxia-inducible factor (HIF)-alpha subunits which play a major role in the transcriptional activation of genes encoding angiogenic factors. HIF-alpha abundance is negatively regulated by a subfamily of dioxygenases referred to as prolyl hydroxylase domain-containing proteins (PHDs) which use O(2) as a substrate to hydroxylate HIF-alpha subunits and hence tag them for rapid degradation. Under hypoxic conditions, HIF-alpha subunits accumulate due to reduced hydroxylation efficiency and form transcriptionally active heterodimers with HIF-1ss to activate the expression of angiogenic factors and other proteins important for cellular adaptation to hypoxia. Angiogenesis is regulated by a combination of at least two different mechanisms. The paracrine mechanism is mediated by non endothelial expression of angiogenic factors such as vascular endothelial growth factor (VEGF)-A, which in turn interact with endothelial cell surface receptors to initiate angiogenic activities. In the autocrine mechanism, endothelial cell themselves are induced to express VEGF-A, which collaborate with the paracrine mechanism to support angiogenesis and protect vascular integrity. Because of critical roles of PHDs and HIFs in regulating angiogenic activities, studies are underway to assess their candidacy as targets for angiogenesis therapies. PMID- 19288063 TI - A vitronectin M381T polymorphism increases risk of hemangioblastoma in patients with VHL gene defect. AB - Hemangioblastomas, highly vascular tumors, occur sporadically or associated with von Hippel-Lindau (VHL) disease. Diverse mutations in the VHL gene inactivate the VHL protein and constitute the molecular etiology of the disease. Changes in VHL gene were analyzed in patients with multiplex ligation-dependent probe amplification and single-strand conformation polymorphism analyses. We report here that other angiogenesis-related changes in vitronectin were identified with 2D electrophoresis of plasma samples and restriction fragment length polymorphisms. Our findings revealed that most patients (80.0%) with a familial VHL deletion carried the threonine (T) allele at vitronectin codon 381. Adults simultaneously carrying a VHL defect and the T allele were 5.0-fold more likely to be affected by VHL disease than were methionine/methionine (M/M) homozygotes carrying a VHL defect. Patients with sporadic hemangioblastoma, C-terminally truncated VHL protein or a large deletion in the VHL gene, and the T allele were 18.0-fold more likely to develop recurrent disease. Taken together, individuals with mutated VHL are more likely to be affected by familial or recurrent sporadic hemangioblastoma when carrying the M/T or T/T genotype at codon 381 of vitronectin. PMID- 19288064 TI - Effect of fiber geometry on pulsatile pumping and energy expenditure. AB - Myocardial fiber orientation is a topic that has recently received much attention in connection with cardiac pumping function. The twisting motion of the cardiac base to apex can be a direct result of this geometric orientation of these fibers. One important question that has not been addressed yet is whether there is any relationship between the contractile energy expenditure and the geometric orientation of myocardial fibers. In the present work, we study the effect of contractile fiber orientation on pumping function. We particularly compare the effect of fiber geometry on ejection fraction, and on the energy required for contraction in both cylindrical and half-ellipsoid shell models. The analytical models we used signify the importance of twisting motion in minimizing the energy required to generate certain ejection fraction. Indeed, we quantified that if the angle of contractile fibers is appropriate for the shape and the size of the pump, twisting scheme can tremendously reduce the energy requirement for pumping. PMID- 19288065 TI - Seasonality and outbreaks in West Nile virus infection. AB - In this paper we analyze the impact of seasonal variations on the dynamics of West Nile Virus infection. We are interested in the generation of new epidemic peaks starting from an endemic state. In many cases, the oscillations generated by seasonality in the dynamics of the infection are too small to be observable. The interplay of this seasonality with the epidemic oscillations can generate new outbreaks starting from the endemic state through a mechanism of parametric resonance. Using experimental data we present specific cases where this phenomenon is numerically observed. PMID- 19288066 TI - Selective backbone labelling of ILV methyl labelled proteins. AB - Adding the 13C labelled 2-keto-isovalerate and 2-oxobutanoate precursors to a minimal medium composed of 12C labelled glucose instead of the commonly used (2D, 13C) glucose leads not only to the 13C labelling of (I, L, V) methyls but also to the selective 13C labelling of the backbone C(alpha) and CO carbons of the Ile and Val residues. As a result, the backbone (1H, 15N) correlations of the Ile and Val residues and their next neighbours in the (i + 1) position can be selectively identified in HN(CA) and HN(CO) planes. The availability of a selective HSQC spectrum corresponding to the sole amide resonances of the Ile and Val residues allows connecting them to their corresponding methyls by the intra-residue NOE effect, and should therefore be applicable to larger systems. PMID- 19288067 TI - Enhanced enzymatic hydrolysis of lignocellulose by optimizing enzyme complexes. AB - To enhance the conversion of the cellulose and hemicellulose, the corncob pretreated by aqueous ammonia soaking was hydrolyzed by enzyme complexes. The saturation limit for cellulase (Spezyme CP) was determined as 15 mg protein/g glucan (50 filter paper unit (FPU)/g glucan). The accessory enzymes (beta glucosidase, xylanase, and pectinase) were supplemented to hydrolyze cellobiose (cellulase-inhibiting product), hemicellulose, and pectin (the component covering the fiber surfaces), respectively. It was found that beta-glucosidase (Novozyme 188) loading of 1.45 mg protein/g glucan [30 cellobiase units (CBU)/g glucan] was enough to eliminate the cellobiose inhibitor, and 2.9 mg protein/g glucan (60 CBU/g glucan) was the saturation limit. The supplementation of xylanase and pectinase can increase the conversion of cellulose and hemicellulose significantly. The yields of glucose and xylose enhanced with the increasing enzyme loading, but the increasing trend became low at high loading. Compared with xylanase, pectinase was more effective to promote the hydrolysis of cellulose and hemicellulose. The supplementation of pectinase with 0.12 mg protein/g glucan could increase the yields of glucose and xylose by 7.5% and 29.3%, respectively. PMID- 19288068 TI - Immobilization of beta-galactosidase onto magnetic beads. AB - A study of the cross-linking of beta-galactosidase on magnetic beads is reported here. The magnetic beads were prepared from artemisia seed gum, chitosan, and magnetic fluid in the presence of a cross-linking regent (i.e., glutaraldehyde). The reactive aldehyde groups of the magnetic beads allowed the reaction of the amino groups of the enzymes. The animated magnetic beads were used for the covalent immobilization of beta-galactosidase. The effect of various preparation conditions on the activity of the immobilized beta-galactosidase, such as immobilizing time, amount of enzyme, and the concentration of glutaraldehyde, were investigated. The influence of pH and temperature on the activity and the stability of the enzyme, both free and immobilized, have been studied. And o nitrophenyl-beta-D: -galactopyranoside (ONPG) was chosen as a substrate. The beta galactosidase immobilized on the magnetic beads resulted in an increase in enzyme stability. Optimum operational temperature for immobilized enzyme was 10 degrees Celsius higher than that of free enzyme and was significantly broader. PMID- 19288069 TI - Biosorption of cadmium, lead, and uranium by powder of poplar leaves and branches. AB - The removal of metal ions from aqueous solutions by biosorption plays an important role in water pollution control. In this study, dried leaves and branches of poplar trees were studied for removing some toxic elements (cadmium, lead, and uranium) from aqueous solutions. The equilibrium experiments were systematically carried out in a batch process, covering various process parameters that include agitation time, adsorbent size and dosage, initial cadmium, lead and uranium concentration, and pH of the aqueous solution. Adsorption behavior was found to follow Freundlich and Langmuir isotherms. The results have shown that both dried leaves and branches can be effectively used for removing uranium, while only branches were found to remove lead and cadmium completely from the aqueous solution. The maximum biosorption capacity of leaves for uranium was found to be 2.3 mg g(-1) and 1.7 mg g(-1) and 2.1 mg g(-1) for lead and cadmium on branches, respectively. In addition, the studied biomass materials were used in removing lead and cadmium from contaminated water and the method was found to be effective. PMID- 19288070 TI - [Discussion of emergency embolization for severe pelvic ring fractures with arterial bleeding]. PMID- 19288071 TI - [Classification of severely injured patients in the G-DRG System 2008]. AB - BACKGROUND: Since the introduction of a per-case reimbursement system in Germany (German Diagnosis-Related Groups, G-DRG), the correct reimbursement for the treatment of severely injured patients has been much debated. While the classification of a patient in a polytrauma DRG follows different rules than the usual clinical definition, leading to a high number of patients not grouped as severely injured by the system, the system was also criticized in 2005 for its shortcomings in financing the treatment of severely injured patients. The development of financial reimbursement will be discussed in this paper. METHOD: 167 patients treated in 2006 and 2007 due to a severe injury at the University Hospital Munster and grouped into a polytrauma-DRG were included in this study. For each patient, cost-equivalents were estimated. For those patients treated in 2007 (n=110), exact costs were calculated following the InEK cost-calculation method. The reimbursement was calculated using the G-DRG-Systems of 2007, 2008 and 2009. Cost-equivalents/costs and clinical parameters were correlated. RESULTS: A total of 167 patients treated in 2006 and 2007 for a severe injury at the Munster University Hospital and grouped into a polytrauma DRG were included in this study. Cost equivalents were estimated for each patient. For those patients treated in 2007 (n=110), exact costs were calculated following the InEK (Institute for the Hospital Remuneration System) cost calculation method. Reimbursement was calculated using the G-DRG systems of 2007, 2008 and 2009. Cost equivalents/costs and clinical parameters were correlated. DISCUSSION: With the ongoing development of the G-DRG system, reimbursement for the treatment of severely injured patient has improved, but the amount of underfinancing remains substantial. As treatment of severely injured patients must be reimbursed using the G-DRG system, this system must be further adapted to better meet the needs of severely injured patients. Parameters such as total surgery time, injury severity score (ISS) and LOS in ICU could be used for this purpose. In future, data obtained in trauma networks can help optimize reimbursement for the treatment of these patients. PMID- 19288072 TI - Indigenous bacteria may interfere with the biocontrol of plant diseases. AB - Prodigiosin is a reddish antibiotic pigment that plays an important role in the biocontrol of plant diseases by the bacterium Serratia marcescens. However, its activity is unstable under agricultural conditions; further, it can be degraded by various environmental factors. To examine the effect of epiphytic microbes on the stability of prodigiosin used for biological control processes, we collected a total of 1,280 bacterial isolates from the phylloplane of cyclamen and tomato plants. Approximately 72% of the bacterial strains isolated from the cyclamen plants and 66% of those isolated from the tomato plants grew on minimal agar medium containing 100 microg ml(-1) prodigiosin. Certain isolates obtained from both plant species exhibited prodigiosin-degrading activity. We compared the 16S rRNA gene sequences derived from the isolates with sequences in a database. The comparison revealed that the sequences determined for the prodigiosin-degrading isolates were homologous to those of the genera Pseudomonas, Caulobacter, Rhizobium, Sphingomonas, Janthinobacterium, Novosphingobium, and Rathayibacter. These results indicate that indigenous epiphytic microorganisms may interfere with the interaction between plant pathogens and biocontrol agents by degrading the antibiotics produced by the agents. PMID- 19288073 TI - Mandible shape and dwarfism in squirrels (Mammalia, Rodentia): interaction of allometry and adaptation. AB - Squirrels include several independent lineages of dwarf forms distributed into two ecological groups: the dwarf tree and flying squirrels. The mandible of dwarf tree squirrels share a highly reduced coronoid process and a condylar process drawn backwards. Dwarf flying squirrels on the other hand, have an elongated coronoid process and a well-differentiated condylar process. To interpret such a difference, Elliptic Fourier Transform was used to evaluate how mandible shape varies with dwarfism in sciurids. The results obtained show that this clear-cut difference cannot be explained by a simple allometric relationship in relation with size decrease. We concluded that the retention of anteriorly positioned eye sockets, in relation with distance estimation, allowed the conservation of a well differentiated coronoid process in all flying species, despite the trend towards its reduction observed among sciurids as their size decreases. PMID- 19288074 TI - Bivariate genetic modelling of the response to an oral glucose tolerance challenge: a gene x environment interaction approach. AB - AIMS/HYPOTHESIS: Twin and family studies have shown the importance of genetic factors influencing fasting and 2 h glucose and insulin levels. However, the genetics of the physiological response to a glucose load has not been thoroughly investigated. METHODS: We studied 580 monozygotic and 1,937 dizygotic British female twins from the Twins UK Registry. The effects of genetic and environmental factors on fasting and 2 h glucose and insulin levels were estimated using univariate genetic modelling. Bivariate model fitting was used to investigate the glucose and insulin responses to a glucose load, i.e. an OGTT. RESULTS: The genetic effect on fasting and 2 h glucose and insulin levels ranged between 40% and 56% after adjustment for age and BMI. Exposure to a glucose load resulted in the emergence of novel genetic effects on 2 h glucose independent of the fasting level, accounting for about 55% of its heritability. For 2 h insulin, the effect of the same genes that already influenced fasting insulin was amplified by about 30%. CONCLUSIONS/INTERPRETATION: Exposure to a glucose challenge uncovers new genetic variance for glucose and amplifies the effects of genes that already influence the fasting insulin level. Finding the genes acting on 2 h glucose independently of fasting glucose may offer new aetiological insight into the risk of cardiovascular events and death from all causes. PMID- 19288075 TI - Established diet-induced obesity in female rats leads to offspring hyperphagia, adiposity and insulin resistance. AB - AIMS/HYPOTHESIS: Accumulating evidence suggests that maternal obesity may increase the risk of metabolic disease in the offspring. We investigated the effects of established maternal diet-induced obesity on male and female offspring appetite, glucose homeostasis and body composition in rats. METHODS: Female Wistar rats were fed either a standard chow (3% fat, 7% sugar [wt/wt]) or a palatable obesogenic diet (11% fat, 43% sugar [wt/wt]) for 8 weeks before mating and throughout pregnancy and lactation. Male and female offspring of control and obese dams were weaned on to standard chow and assessed until 12 months of age. RESULTS: At mating, obese dams were heavier than control with associated hyperglycaemia and hyperinsulinaemia. Male and female offspring of obese dams were hyperphagic (p < 0.0001) and heavier than control (p < 0.0001) until 12 months of age. NEFA were raised at 2 months but not at 12 months. At 3 months, OGTT showed more pronounced alteration of glucose homeostasis in male than in female offspring of obese animals. Euglycaemic-hyperinsulinaemic clamps performed at 8 to 9 months in female and 10 to 11 months in male offspring revealed insulin resistance in male offspring of obese dams (p < 0.05 compared with control). Body compositional analysis at 12 months also showed increased fat pad weights in male and female offspring of obese animals. CONCLUSIONS/INTERPRETATION: Diet-induced obesity in female rats leads to a state of insulin resistance in male offspring, associated with development of obesity and increased adiposity. An increase in food intake may play a role. PMID- 19288076 TI - Regulation of insulin secretion: a matter of phase control and amplitude modulation. AB - The consensus model of stimulus-secretion coupling in beta cells attributes glucose-induced insulin secretion to a sequence of events involving acceleration of metabolism, closure of ATP-sensitive K(+) channels, depolarisation, influx of Ca(2+) and a rise in cytosolic free Ca(2+) concentration ([Ca(2+)](c)). This triggering pathway is essential, but would not be very efficient if glucose did not also activate a metabolic amplifying pathway that does not raise [Ca(2+)](c) further but augments the action of triggering Ca(2+) on exocytosis. This review discusses how both pathways interact to achieve temporal control and amplitude modulation of biphasic insulin secretion. First-phase insulin secretion is triggered by the rise in [Ca(2+)](c) that occurs synchronously in all beta cells of every islet in response to a sudden increase in the glucose concentration. Its time course and duration are shaped by those of the Ca(2+) signal, and its amplitude is modulated by the magnitude of the [Ca(2+)](c) rise and, substantially, by amplifying mechanisms. During the second phase, synchronous [Ca(2+)](c) oscillations in all beta cells of an individual islet induce pulsatile insulin secretion, but these features of the signal and response are dampened in groups of intrinsically asynchronous islets. Glucose has hardly any influence on the amplitude of [Ca(2+)](c) oscillations and mainly controls the time course of triggering signal. Amplitude modulation of insulin secretion pulses largely depends on the amplifying pathway. There are more similarities than differences between the two phases of glucose-induced insulin secretion. Both are subject to the same dual, hierarchical control over time and amplitude by triggering and amplifying pathways, suggesting that the second phase is a sequence of iterations of the first phase. PMID- 19288077 TI - The T allele of rs7903146 TCF7L2 is associated with impaired insulinotropic action of incretin hormones, reduced 24 h profiles of plasma insulin and glucagon, and increased hepatic glucose production in young healthy men. AB - AIMS/HYPOTHESIS: We studied the physiological, metabolic and hormonal mechanisms underlying the elevated risk of type 2 diabetes in carriers of TCF7L2 gene. METHODS: We undertook genotyping of 81 healthy young Danish men for rs7903146 of TCF7L2 and carried out various beta cell tests including: 24 h glucose, insulin and glucagon profiles; OGTT; mixed meal test; IVGTT; hyperglycaemic clamp with co infusion of glucagon-like peptide (GLP)-1 or glucose-dependent insulinotropic polypeptide (GIP); and a euglycaemic-hyperinsulinaemic clamp combined with glucose tracer infusion to study hepatic and peripheral insulin action. RESULTS: Carriers of the T allele were characterised by reduced 24 h insulin concentrations (p < 0.05) and reduced insulin secretion relative to glucose during a mixed meal test (beta index: p < 0.003), but not during an IVGTT. This was further supported by reduced late-phase insulinotropic action of GLP-1 (p = 0.03) and GIP (p = 0.07) during a 7 mmol/l hyperglycaemic clamp. Secretion of GLP 1 and GIP during the mixed meal test was normal. Despite elevated hepatic glucose production, carriers of the T allele had significantly reduced 24 h glucagon concentrations (p < 0.02) suggesting altered alpha cell function. CONCLUSIONS/INTERPRETATION: Elevated hepatic glucose production and reduced insulinotropic effect of incretin hormones contribute to an increased risk of type 2 diabetes in carriers of the rs7903146 risk T allele of TCF7L2. PMID- 19288078 TI - Mechanical ventilation: quo vadis? PMID- 19288079 TI - Epidemiology of mechanical ventilation: analysis of the SAPS 3 database. AB - OBJECTIVE: To evaluate current practice of mechanical ventilation in the ICU and the characteristics and outcomes of patients receiving it. DESIGN: Pre-planned sub-study of a multicenter, multinational cohort study (SAPS 3). PATIENTS: 13,322 patients admitted to 299 intensive care units (ICUs) from 35 countries. INTERVENTIONS: None. MAIN MEASUREMENTS AND RESULTS: Patients were divided into three groups: no mechanical ventilation (MV), noninvasive MV (NIV), and invasive MV. More than half of the patients (53% [CI: 52.2-53.9%]) were mechanically ventilated at ICU admission. FIO2, VT and PEEP used during invasive MV were on average 50% (40-80%), 8 mL/kg actual body weight (6.9-9.4 mL/kg) and 5 cmH2O (3-6 cmH2O), respectively. Several invMV patients (17.3% (CI:16.4-18.3%)) were ventilated with zero PEEP (ZEEP). These patients exhibited a significantly increased risk-adjusted hospital mortality, compared with patients ventilated with higher PEEP (O/E ratio 1.12 [1.05-1.18]). NIV was used in 4.2% (CI: 3.8 4.5%) of all patients and was associated with an improved risk-adjusted outcome (OR 0.79, [0.69-0.90]). CONCLUSION: Ventilation mode and parameter settings for MV varied significantly across ICUs. Our results provide evidence that some ventilatory modes and settings could still be used against current evidence and recommendations. This includes ventilation with tidal volumes >8mL/kg body weight in patients with a low PaO2/FiO2 ratio and ZEEP in invMV patients. Invasive mechanical ventilation with ZEEP was associated with a worse outcome, even after controlling for severity of disease. Since our study did not document indications for MV, the association between MV settings and outcome must be viewed with caution. PMID- 19288080 TI - Anatomic ACL reconstruction: does the platelet-rich plasma accelerate tendon healing? AB - Recently, the use of hamstring tendons in anterior cruciate ligament repair has been increasing. However, tendon-to-bone healing occurs slowly, which can be a problem to an early return to sport activities. The use of growth factors from platelets seems to improve tissue healing. We enrolled 40 patients in a prospective study that were submitted to an anatomic reconstruction of the anterior cruciate ligament. Patients were sequentially enrolled into four groups: group A without platelet-rich plasma (PRP); group B with PRP in femoral tunnels at the end of surgery; group C with PRP in femoral tunnels at the end of surgery and intra-articular at 2- and 4 weeks after surgery; group D with PRP activated with thrombin in the femoral tunnels. All patients underwent magnetic resonance imaging of the knee 3 months after surgery to evaluate the signal intensity of the fibrous interzone (FIZ) in the femoral tunnels. We did not find any difference among the groups when comparing the signal intensity of the FIZ on magnetic resonance imaging. PMID- 19288082 TI - Endoscopic treatment of chronic mid-portion Achilles tendinopathy: novel technique with short-term results. AB - In this prospective study, we present the short-term clinical results of a novel endoscopic surgical technique for patients suffering from chronic painful mid portion Achilles tendinopathy. Eight consecutive patients (seven men and one woman, mean age 52 years) diagnosed with chronic painful mid-portion Achilles tendinopathy were included in this study and were treated with endoscopic debridement of the ventral neovascularized area, the peritendineum and the Achilles tendon. Patients recorded the function of the Achilles tendon and the severity of Achilles tendon pain during tendon loading activity, pre and postoperatively in a visual analogue scale. Patient global satisfaction was also assessed in a similar manner. Patients were followed-up for 6 months. All patients experienced immediate postoperative pain relief. In terms of Achilles tendon pain, the median visual analogue score (VAS) increased from 40 (10-60) (preoperatively) to 97.5 (85-100) (last follow-up examination). In terms of Achilles tendon function, the median VAS increased from 22.5 (0-30) (preoperatively) to 90 (80-95) (last follow-up examination). In terms of global satisfaction, the median VAS in the last follow-up examination was 85 (70-95). No postoperative complications were recorded. In conclusion, the short-term clinical results were satisfactory. PMID- 19288083 TI - Review: femoral tunnel placement for PCL reconstruction in relation to the PCL fibre bundle attachments. AB - Accurate placement of the femoral tunnel position is an important aspect of cruciate ligament reconstruction surgery. However, a wide range of positions have been described in the literature for the femoral tunnels for posterior cruciate ligament (PCL) reconstruction, and there is no consensus regarding the optimal placement. It was hypothesised that some of the prior studies used femoral PCL graft tunnels that were outside of the anatomical attachments. A systematic review and descriptive anatomical study in vitro were undertaken. Fifty articles that described PCL reconstruction since 1917 were reviewed; the description of the femoral tunnel placement was reproducible from only 20 of these. A right, distal femur was disarticulated and the PCL attachment was dissected; the centres of the anterolateral and posteromedial bundles were marked and confirmed to be close to the mean published data. The tunnel positions described in the 20 papers were also marked. The descriptions of 15 of the 20 anterolateral and 4 of the 5 posteromedial bundle tunnel centres were close (within +/-2 mm deep and +/-01:00 o'clock) to the anatomical centres of the bundle attachments. Other tunnels were placed either deeper or shallower than the anatomical attachments. The mean positions were: 7 mm from the articular cartilage at 1 o'clock for the anterolateral bundle, and 8 mm from the cartilage at 3 o'clock for the posteromedial bundle, in a right knee. It was concluded that it was not possible to replicate the tunnel positions used in most of the literature because of inadequate descriptions. Most of the usable descriptions of the anterolateral bundle position were close to the anatomical centre, but some tunnels were placed non-anatomically. A reproducible system for description of the femoral tunnel positions should be used. PMID- 19288084 TI - Comparison of ketanserin, buspirone and propranolol on arousal, pupil size and autonomic function in healthy volunteers. AB - RATIONALE: The human pupil may be a suitable physiological test system for the assessment of excessive daytime sleepiness (EDS), but pupillometric assessment could be confounded by medication for comorbid hypertension and mood disorders. OBJECTIVES: We examined the profile of the 5HT-2/alpha1/H1 antagonist ketanserin, the 5HT1a agonist buspirone and the beta adrenoceptor antagonist propranolol on pupillary and other measures of arousal. MATERIALS AND METHODS: Ketanserin (20 mg), buspirone (10 mg) and propranolol (40 mg) were administered in three independent experiments according to a crossover, placebo-controlled, double blind design. Resting pupil diameter (RPD) was sampled over 5-min in darkness with infrared pupillometry. Tests also included critical flicker fusion frequency (CFFF), visual analogue scales (VAS), the pupillary light reflex and heart rate/blood pressure. RESULTS: Ketanserin reduced RPD, CFFF, VAS-rated arousal and blood pressure and increased the light reflex amplitude. Buspirone reduced RPD and blood pressure. Propranolol reduced heart rate but had no effects on pupillary functions or any arousal measure. CONCLUSIONS: Ketanserin but not propranolol had a fully sedative profile and may confound pupillometric assessment of EDS. Beta adrenergic receptors do not appear to participate in arousal and pupillary functions, while 5HT1a receptors reduce pupil size without affecting arousal. Pupil size may not be used unequivocally as an index of the level of alertness in the case of drug-induced changes, when drugs interfere with the central pupil control mechanism in ways that are unrelated to their effects on arousal. PMID- 19288085 TI - Comparison of the discriminative stimulus effects of dimethyltryptamine with different classes of psychoactive compounds in rats. AB - RATIONALE: There has been increased recreational use of dimethyltryptamine (DMT), but little is known of its discriminative stimulus effects. OBJECTIVES: The present study assessed the similarity of the discriminative stimulus effects of DMT to other types of hallucinogens and to psychostimulants. METHODS: Rats were trained to discriminate DMT from saline. To test the similarity of DMT to known hallucinogens, the ability of (+)-lysergic acid diethylamide (LSD), (-)-2,5 dimethoxy-4-methylamphetamine (DOM), (+)-methamphetamine, or (+/-)3,4 methylenedioxymethyl amphetamine (MDMA) to substitute in DMT-trained rats was tested. The ability of DMT to substitute in rats trained to discriminate each of these compounds was also tested. To assess the degree of similarity in discriminative stimulus effects, each of the compounds was tested for substitution in all of the other training groups. RESULTS: LSD, DOM, and MDMA all fully substituted in DMT-trained rats, whereas DMT fully substituted only in DOM trained rats. Full cross-substitution occurred between DMT and DOM, LSD and DOM, and (+)-methamphetamine and MDMA. MDMA fully substituted for (+)-methamphetamine, DOM, and DMT, but only partially for LSD. In MDMA-trained rats, LSD and (+) methamphetamine fully substituted, whereas DMT and DOM did not fully substitute. No cross-substitution was evident between (+)-methamphetamine and DMT, LSD, or DOM. CONCLUSIONS: DMT produces discriminative stimulus effects most similar to those of DOM, with some similarity to the discriminative stimulus effects of LSD and MDMA. Like DOM and LSD, DMT seems to produce predominately hallucinogenic like discriminative stimulus effects and minimal psychostimulant effects, in contrast to MDMA which produced hallucinogen- and psychostimulant-like effects. PMID- 19288086 TI - Nanofluidics. PMID- 19288087 TI - Mendeleyev vodka challenge. PMID- 19288088 TI - Internal correction of hafnium oxide spectral interferences and mass bias in the determination of platinum in environmental samples using isotope dilution analysis. AB - A method has been developed for the accurate determination of platinum by isotope dilution analysis, using enriched (194)Pt, in environmental samples containing comparatively high levels of hafnium without any chemical separation. The method is based on the computation of the contribution of hafnium oxide as an independent factor in the observed isotope pattern of platinum in the spiked sample. Under these conditions, the ratio of molar fractions between natural abundance and isotopically enriched platinum was independent of the amount of hafnium present in the sample. Additionally, mass bias was corrected by an internal procedure in which the regression variance was minimised. This was possible as the mass bias factor for hafnium oxide was very close to that of platinum. The final procedure required the measurement of three platinum isotope ratios (192/194, 195/194 and 196/194) to calculate the concentration of platinum in the sample. The methodology has been validated using the reference material "BCR-723 road dust" and has been applied to different environmental matrices (road dust, air particles, bulk wet deposition and epiphytic lichens) collected in the Aspe Valley (Pyrenees Mountains). A full uncertainty budget, using Kragten's spreadsheet method, showed that the total uncertainty was limited only by the uncertainty in the measured isotope ratios and not by the uncertainties of the isotopic composition of platinum and hafnium. PMID- 19288090 TI - On-line corrections for visuomotor errors. AB - This study was designed to determine how visual feedback mediates error corrections during reaching. We used visuomotor rotations to dissociate a cursor, representing finger position, from the actual finger location. We then extinguished cursor feedback at different distances from the start location to determine whether corrections were based on error extrapolation from prior cursor information. Results indicated that correction amplitude varied with the extent of cursor feedback. A second experiment tested specific aspects of error information that might mediate corrections to visuomotor rotations: rotation angle, distance between the finger and cursor positions and the duration of cursor exposure. Results showed that corrections did not depend on the amplitude of the rotation angle or the amount of time the cursor was shown. Instead, participants corrected for the cursor-finger distance, at the point where cursor feedback was last-seen. These findings suggest that within-trial corrections and inter-trial adaptation might employ different mechanisms. PMID- 19288089 TI - Migraine: where and how does the pain originate? AB - Migraine is a complex neurological disease with a genetic background. Headache is the most prominent and clinically important symptom of migraine but its origin is still enigmatic. Numerous clinical, histochemical, electrophysiological, molecular and genetical approaches form a puzzle of findings that slowly takes shape. The generation of primary headaches like migraine pain seems to be the consequence of multiple pathophysiological changes in meningeal tissues, the trigeminal ganglion, trigeminal brainstem nuclei and descending inhibitory systems, based on specific characteristics of the trigeminovascular system. This contribution reviews the current discussion of where and how the migraine pain may originate and outlines the experimental work to answer these questions. PMID- 19288091 TI - Pulmonary infections in HIV-positive children. AB - Infection of the lungs and airways by viral, bacterial, fungal and protozoal agents, often producing atypical radiographic features, is common in children with human immunodeficiency virus (HIV) infection. Conventional chest radiography and chest CT remain the most useful imaging modalities for evaluation of the immunocompromised patient presenting with a suspected pulmonary infection. In this review the radiological features of acute lung infections in this population are discussed. PMID- 19288092 TI - Identification of an additional two-cysteine containing type I interferon in rainbow trout Oncorhynchus mykiss provides evidence of a major gene duplication event within this gene family in teleosts. AB - Multiple type I interferons (IFNs) have recently been identified in salmonids, containing two or four conserved cysteines. In this work, a novel two-cysteine containing (2C) IFN gene was identified in rainbow trout. This novel trout IFN gene (termed IFN5) formed a phylogenetic group that is distinct from the other three salmonid IFN groups sequenced to date and had a close evolutionary relationship with IFNs from advanced fish species. Our data demonstrate that two subgroups are apparent within each of the 2C and 4C type I IFNs, an evolutionary outcome possibly due to two rounds of genome duplication events that have occurred within teleosts. We have examined gene expression of the trout 2C type I IFN in cultured cells following stimulation with lipopolysaccharide, phytohaemagglutinin, polyI:C or recombinant IFN, or after transfection with polyI:C. The kinetics of gene expression was also studied after viral infection. Analysis of the regulatory elements in the IFN promoter region predicted several binding sites for key transcription factors that potentially play an important role in mediating IFN5 gene expression. PMID- 19288093 TI - A novel protease-resistant alpha-galactosidase with high hydrolytic activity from Gibberella sp. F75: gene cloning, expression, and enzymatic characterization. AB - A novel alpha-galactosidase gene (aga-F75) from Gibberella sp. F75 was cloned and expressed in Escherichia coli. The gene codes for a protein of 744 amino acids with a 24-residue putative signal peptide and a calculated molecular mass of 82.94 kDa. The native structure of the recombinant Aga-F75 was estimated to be a trimer or tetramer. The deduced amino acid sequence showed highest identity (69%) with an alpha-galactosidase from Hypocrea jecorina (Trichoderma reesei), a member of the glycoside hydrolase family 36. Purified recombinant Aga-F75 was optimally active at 60 degrees C and pH 4.0 and was stable at pH 3.0-12.0. The enzyme exhibited broad substrate specificity and substantial resistance to neutral and alkaline proteases. The enzyme K (m) values using pNPG, melibiose, stachyose, and raffinose as substrates were 1.06, 1.75, 54.26, and 8.23 mM, respectively. Compared with the commercial alpha-galactosidase (Aga-A) from Aspergillus niger var. AETL and a protease-resistant alpha-galactosidase (Aga-F78) from Rhizopus sp. F78, Aga-F75 released 1.4- and 4.9-fold more galactose from soybean meal alone, respectively, and 292.5- and 8.6-fold more galactose from soybean meal in the presence of trypsin, respectively. The pH and thermal stability and hydrolytic activity of Aga-F75 make it potentially useful in the food and feed industries. PMID- 19288094 TI - Efficient synthesis of enantiomeric ethyl lactate by Candida antarctica lipase B (CALB)-displaying yeasts. AB - The whole-cell biocatalyst displaying Candida antarctica lipase B (CALB) on the yeast cell surface with alpha-agglutinin as the anchor protein was easy to handle and possessed high stability. The lyophilized CALB-displaying yeasts showed their original hydrolytic activity and were applied to an ester synthesis using ethanol and L: -lactic acid as substrates. In water-saturated heptane, CALB-displaying yeasts catalyzed ethyl lactate synthesis. The synthesis efficiency increased depending on temperature and reached approximately 74% at 50 degrees C. The amount of L: -ethyl lactate increased gradually. L: -Ethyl lactate synthesis stopped at 200 h and restarted after adding of L: -lactic acid at 253 h. It indicated that CALB-displaying yeasts retained their synthetic activity under such reaction conditions. In addition, CALB-displaying yeasts were able to recognize L: -lactic acid and D: -lactic acid as substrates. L: -Ethyl lactate was prepared from L: -lactic acid and D: -ethyl lactate was prepared from D: lactic acid using the same CALB-displaying whole-cell biocatalyst. These findings suggest that CALB-displaying yeasts can supply the enantiomeric lactic esters for preparation of useful and improved biopolymers of lactic acid. PMID- 19288095 TI - The "rising bubble" sign: a new aid in the diagnosis of unicameral bone cysts. AB - The observation of a bubble of gas at the most non-dependent margin of a lytic bone lesion which has sustained a pathologic fracture implies that the lesion is hollow and can assist the radiologist in making the diagnosis of a unicameral bone cyst. The imaging studies of two patients who sustained pathologic fractures through unicameral bone cysts and exhibited the "rising bubble" sign are shown. The sign's basis, proper utilization, and potential pitfalls are discussed. PMID- 19288096 TI - Synthesis and preclinical evaluation of [(11)C]PAQ as a PET imaging tracer for VEGFR-2. AB - PURPOSE: (R,S)-N-(4-Bromo-2-fluorophenyl)-6-methoxy-7-((1-methyl-3 piperidinyl)methoxy)-4-quinazolinamine (PAQ) is a tyrosine kinase inhibitor with high affinity for the vascular endothelial growth factor receptor 2 (VEGFR-2), which plays an important role in tumour angiogenesis. The aim of this work was to develop and evaluate in mice the (11)C-labelled analogue as an in vivo tracer for VEGFR-2 expression in solid tumours. METHODS: [(11)C]PAQ was synthesized by an N methylation of desmethyl-PAQ using [(11)C]methyl iodide. The tracer's pharmacokinetic properties and its distribution in both subcutaneous and intraperitoneal tumour models were evaluated with positron emission tomography (PET). [(18)F]FDG was used as a reference tracer for tumour growth. PET results were corroborated by ex vivo and in vitro phosphor imaging and immunohistochemical analyses. RESULTS: In vitro assays and PET in healthy animals revealed low tracer metabolism, limited excretion over 60 min and a saturable and irreversible binding. Radiotracer uptake in subcutaneous tumour masses was low, while focal areas of high uptake (up to 8% ID/g) were observed in regions connecting the tumour to the host. Uptake was similarly high but more distributed in tumours growing within the peritoneum. The pattern of radiotracer uptake was generally different from that of the metabolic tracer [(18)F]FDG and correlated well with variations in VEGFR-2 expression determined ex vivo by immunohistochemical analysis. CONCLUSION: These results suggest that [(11)C]PAQ has potential as a noninvasive PET tracer for in vivo imaging of VEGFR-2 expression in angiogenic "hot spots". PMID- 19288097 TI - Nanoparticles for concurrent multimodality imaging and therapy: the dawn of new theragnostic synergies. PMID- 19288098 TI - Intraoperative radioguidance with a portable gamma camera: a novel technique for laparoscopic sentinel node localisation in urological malignancies. AB - PURPOSE: Our aim was to assess the feasibility of intraoperative radioguidance with a portable gamma camera during laparoscopic sentinel node (SN) procedures in urological malignancies. METHODS: We evaluated the use of the intraoperative portable gamma camera in 20 patients: 16 patients with prostate carcinoma (PCC), 2 patients with renal cell carcinoma (RC) and 2 patients with testicular cancer (TC). Intra/peritumoural injection of (99m)Tc-nanocolloid ((99m)Tc) was followed by planar lymphoscintigraphy, SPECT/CT and marking of SN levels. Before laparoscopy a (125)I seed was fixed on the laparoscopic gamma probe as a pointer of SN seeking. The portable gamma camera was set to display the (99m)Tc signal for SN localisation and the (125)I signal for SN seeking. Matching of these signals on screen indicated exact SN localisation, and consequently this SN was removed. RESULTS: The mean injected dose was 218 MBq in PCC, 228 MBq in RC and 88 MBq in TC. Pelvic SN were visualised in all PCC patients, with uncommonly located SN in seven patients. SN metastases were found in seven patients (one in a uncommonly located SN). Both RC patients and TC patients had para-aortic SN, which were all tumour free. A total of 59 SN were removed. The portable gamma camera enabled real-time SN display/identification in 18 patients (90%). CONCLUSION: The use of a portable gamma camera in combination with a laparoscopic gamma probe incorporates intraoperative real-time imaging with improved SN identification in urological malignancies. This procedure might also be useful for SN identification of other deep draining malignancies. PMID- 19288099 TI - Optimization and assessment of quantitative 124I imaging on a Philips Gemini dual GS PET/CT system. AB - PURPOSE: Quantitative (124)I PET imaging is challenging as (124)I has a complex decay scheme. In this study the performance of a Philips Gemini dual GS PET/CT system was optimized and assessed for (124)I. METHODS: The energy window giving the maximum noise equivalent count rate (NECR) and NEMA 2001-NU2 image quality were measured. The activity concentration (AC) accuracy of images calibrated using factors from (18)F and (124)I decaying source measurements were investigated. RESULTS: The energy window 455-588 keV gave the maximum NECR of 9.67 kcps for 233 MBq. (124)I and (18)F image quality was comparable, although (124)I background variability was increased. The average underestimation in AC in (124)I images was 17.9 +/- 2.9% for nonuniform background and 14.7 +/- 2.9% for single scatter simulation (SSS) subtraction scatter correction. At 224 MBq the underestimation was 10.8 +/- 11.3%, which is comparable to 7.7 +/- 5.3% for (18)F, but increased with decreasing activity. CONCLUSIONS: The best (124)I PET quantitative accuracy was achieved for the optimized energy window, using SSS scatter correction and calibration factors from decaying (124)I source measurements. The quantitative accuracy for (124)I was comparable to that for (18)F at high activities of 224 MBq but diminishing with decreasing activity. Specific corrections for prompt gamma-photons may further improve the quantitative accuracy. PMID- 19288100 TI - Radiopharmaceutical clinical trials - dose is more than sieverts and becquerels. PMID- 19288101 TI - The effect of aging on OX40 agonist-mediated cancer immunotherapy. AB - Agents that enhance T cell co-stimulatory signaling have emerged as promising cancer immunotherapies. Our laboratory has been evaluating the TNF receptor co stimulatory molecule, OX40, which has the capacity to augment critical aspects of T cell function and induce tumor regression in animal models. Effective stimulation of OX40 expressing T cells was accomplished with agonist antibodies to OX40 that were eventually translated into a clinical trial for cancer patients. A recent attempt to assess the affect of immune senescence on OX40 therapy, revealed a dramatic loss of efficacy of the agonist therapy in older tumor-bearing mice. The deficiency in OX40-enhanced anti-tumor responses in older mice correlated with a decrease in the number of differentiated effector T cells. Further investigation suggests that the underlying age-related decline in the agonist OX40-mediated T cell responses was not inherent to the T cells themselves, but related to the host environment. Thus, effective use of immunotherapies based on T cell co-stimulatory molecules may require additional modifications, such as immune stimulants to increase innate immunity, to address age-related defects that reside outside of the T cell and within the host environment. PMID- 19288102 TI - Unstable trochanteric fractures: the role of lateral wall reconstruction. AB - The sliding compression device, a widely used implant in unstable proximal femoral fractures, suffers from two major limitations: excessive collapse and screw cut-out. Commonly attributed reasons for these are lateral wall comminution and single-point fixation, respectively. We report our experience of stabilising 74 unstable trochanteric fractures, of which 46 cases underwent lateral wall reconstruction using a trochanteric stabilising plate (TSP) in combination with a dynamic hip screw (DHS), and 34 cases with an intact lateral wall had a DHS with an additional anti-rotation screw providing two-point fixation. Fracture consolidation was observed in all cases at an average of 13.56 weeks. Overall functional hip score as per the Salvati and Wilson scoring system was >30 points in 55 patients. Lateral wall reconstruction is an important component in stabilisation of unstable trochanteric fractures and a combination of TSP with a DHS appears to be a useful device to achieve this. Addition of an antirotation screw is likely to enhance the stability further by providing two-point fixation. PMID- 19288103 TI - Treatment outcome of adult patients with Burkitt lymphoma: results using the LMB protocol in Korea. AB - Burkitt lymphoma (BL) is a rare subtype of adult non-Hodgkin lymphoma, so studies on the outcome of adult BL, especially in Asian patients, are scarce. We report our results using the LMB protocol on Korean adult BL patients. Thirty-eight newly diagnosed BL patients were treated with the LMB protocol; 29 males and nine females with a median age of 47 years (range 18-70) were analyzed, and 14 (36.8%) patients had central nervous system or bone marrow involvement. After the induction phase, 28 patients achieved complete response (CR, 73.7%) and five showed partial response (PR, 13.2%). Among those achieving CR, only four showed relapse. All of the non-CR patients died, including five PR and one with progressive disease. The other four patients died because of infection after the first course of induction. The progression-free and overall estimated survival at 5 years was 74.99% and 68.10%, respectively. Of the 12 patients who died, the median survival was 4.43 months (95% confidence interval 1.43-7.43 months). Thus, most deaths occurred shortly after diagnosis, and four patients older than 58 years died after the first induction cycle. Most early deaths were caused by treatment-related morbidity and failure to achieve complete response. B symptoms, advanced age, bone marrow involvement, and St. Jude/Murphy stage IV classification were significantly associated with poor overall survival. In conclusion, the LMB protocol was effective for Korean adult BL patients. However, considering the high incidence of treatment-related deaths and the poor outcome of non-CR patients, risk-adapted modification of the induction phase is warranted. PMID- 19288104 TI - Topoisomerase I involvement in schedule-dependent interaction between 5-fluoro uracil and irinotecan in the treatment of colorectal cancer. AB - 5-Fluoro-uracil (5FU), an antimetabolite drug, stimulates expression of topoisomerase I (tpI) in adenocarcinoma cancer cells. When 5FU is given in combination with Irinotecan (IR), a tpI poison, the most effective regimen is represented by IR given before low doses of 5FU. Hence, despite their distinct mechanisms of action, the molecular basis for successful combination and schedule of 5FU and IR in the treatment of colorectal cancer rests on the opposing drug effects on the expression and poisoning of the tpI enzyme. PMID- 19288105 TI - Lack of large intragenic rearrangements in dihydropyrimidine dehydrogenase (DPYD) gene in fluoropyrimidine-treated patients with high-grade toxicity. AB - PURPOSE: Deficiency of dihydropyrimidine dehydrogenase (DPD) has been associated with severe fluoropyrimidines (FP) toxicity. Mutations in DPD-coding gene (DPYD) were shown to increase the risk of severe toxicity in FP-treated cancer patients. However, the majority of DPYD alterations characterized in these patients has been considered as polymorphisms and known deleterious mutations are rare and present in only limited subgroup of patients with high toxicity. Recently, the common fragile site FRA1E was mapped within DPYD locus but intragenic rearrangements in DPYD gene were not studied so far. METHODS: We performed the analysis of intragenic rearrangements of DPYD using multiplex ligation-dependent probe amplification in 68 patients with high-grade gastrointestinal and/or hematological toxicity developed at the beginning of FP treatment. RESULTS: We did not detect any deletion/duplication of one or more DPYD exons in analyzed patients. CONCLUSIONS: We assume that rearrangements in DPYD gene play insignificant role in the development of serious FP-related toxicity. PMID- 19288106 TI - A multiple timescale analysis of a mathematical model of the Wnt/beta-catenin signalling pathway. AB - The Wnt signalling pathway is involved in stem cell maintenance, differentiation and tissue development. Its disregulation has also been implicated in many cancers. Beta-catenin is a protein that regulates both transcription of many genes and cell-cell adhesion; in response to an external Wnt stimulus the intracellular levels of beta-catenin are controlled by the proteins which make up the Wnt/beta-catenin signalling pathway. In this paper we present a systematic asymptotic analysis of an existing model of the Wnt signalling pathway due to Lee et al. (PLoS Biol 1:116-132, 2003), highlighting the operation of different pathway components over different timescales. Guided by this analysis we derive a simplified model which is shown to retain the essential behaviour of the full Wnt pathway, recreating the accumulation and degradation of beta-catenin in response to a Wnt stimulus. PMID- 19288107 TI - Improvements in the production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture. AB - The objective of this study was to produce durum wheat doubled haploid (DH) plants through the induction of microspore embryogenesis. The microspore culture technique was improved to maximize production of green plants per spike using three commercial cultivars. Studies on factors such as induction media composition, induction media support and the stage and growth of donor plants were carried out in order to develop an efficient protocol to regenerate green and fertile DH plants. Microspores were plated on a C(17) induction culture medium with ovary co-culture and a supplement of glutathione plus glutamine; 300 g/l Ficoll Type-400 was incorporated to the induction medium support. Donor plants were fertilized with a combination of macro and microelements. With the cultivars 'Ciccio' and 'Claudio' an average of 36.5 and 148.5 fertile plants were produced, respectively, from 1,000 anthers inoculated. This technique was then used to produce fertile DH plants of potential agronomic interest from a collection of ten F(1) crosses involving cultivars of high breeding value. From these crosses 849 green plants were obtained and seed was harvested from 702 plants indicating that 83% of green plants were fertile and therefore were spontaneously DHs. No aneuploid plant was obtained. The 702 plants yielded enough seeds to be field tested. One of the DH lines obtained by microspore embryogenesis, named 'Lanuza', has been sent to the Spanish Plant Variety Office for Registration by the Batlle Seed Company. This protocol can be used instead of the labor-intensive inter-generic crossing with maize as an economically feasible method to obtain DHs for most crosses involving the durum wheat cultivars grown in Spain. PMID- 19288108 TI - Sequence analysis and functional characterization of the promoter of the Picea glauca Cinnamyl Alcohol Dehydrogenase gene in transgenic white spruce plants. AB - The enzyme Cinnamyl Alcohol Dehydrogenase (CAD) catalyses the last step of lignin monomer synthesis, and is considered as a molecular marker of cell wall lignification in different plants species. Here, we report the isolation and analysis of 5' flanking genomic DNA regions upstream to the CAD gene, from two conifers, i.e. white spruce (Picea glauca (Moench) Voss) and loblolly pine (Pinus taeda L.). Sequence comparisons with available CAD gene promoters from angiosperms highlighted the conservation of cis-elements matching MYB, WRKY and bHLH binding sites. Functional characterization of the P. glauca CAD promoter used P. glauca seedlings stably transformed with a DNA fragment of 1,163 base pairs (PgCAD) fused to the beta-glucuronidase (GUS) gene. Histochemical observations of different vegetative organs of the transgenic trees showed that this sequence was sufficient to drive GUS expression in lignifying tissues, and more specifically in differentiating xylem cells. Quantitative RT-PCR experiments also indicated that the native CAD gene was preferentially expressed in differentiating xylem both in stems and roots. In addition, GUS expression driven by the PgCAD promoter was wound-inducible which was consistent with the accumulation of CAD mRNA in response to jasmonate application and mechanical wounding. The spruce CAD promoter represents a valuable tool for research and biotechnology applications related to xylem and wood. PMID- 19288110 TI - Abstracts of the European Congress of Radiology 2009. March 6-9, 2009. Vienna, Austria. PMID- 19288109 TI - Diffusion-weighted imaging (DWI) in MR mammography (MRM): clinical comparison of echo planar imaging (EPI) and half-Fourier single-shot turbo spin echo (HASTE) diffusion techniques. AB - Diffusion-weighted imaging (DWI) techniques have shown potential to differentiate between benign and malignant neoplasms. However, the diagnostic significance of using DWI under routine conditions remains unclear. This study investigated the use of echo planar imaging (EPI) and half-Fourier acquired single-shot turbo spin echo (HASTE)-DWI with respect to the three parameters: lesion visibility, apparent diffusion coefficient (ADC) measurements, and size estimation. Following MRM (1.5 T), EPI- and HASTE-DWI were applied in 65 patients. Lesion visibility on DWI was compared with lesion visibility on subtracted contrast-enhanced T1w images (CE-T1w). Statistical tests were applied to diameter, visibility, and ADC value measurements. Seventy-four lesions were identified. ADC value measurements did not differ significantly between the two DWI sequences. The sensitivity and specificity of routine diagnostics (97.4% and 85.7%) were superior to EPI-DWI (87.2% and 82.9%) and HASTE-DWI (76.9% and 88.6%). Selecting only nonmass lesions, DWI did not prove to be of diagnostic value. Lesion demarcation by DWI was significantly lower compared with that by CE-T1w, with EPI-DWI showing the better performance (p < 0.001). No significant differences were found for size measurements between CE-T1w and DWI. Although clearly inferior compared with CE T1w imaging, both DWI techniques are applicable for lesion assessment and size measurements. PMID- 19288118 TI - Gastroschisis: a third world perspective. AB - PURPOSE: The incidence of gastroschisis is rising worldwide. In developed countries advances in neonatal intensive care (NICU) and the availability of total parenteral nutrition have improved survival for such patients, but in the third world mortality rates remain high. The aim of this study was to evaluate the impact of modern intensive care facilities on the mortality of babies with gastroschisis in Africa. METHODS: A retrospective review of all neonates admitted with a diagnosis of gastroschisis at Inkosi Albert Luthuli Central Hospital in Durban over a 6-year period (2002-2007) was conducted. RESULTS: A total of 106 babies with gastroschisis presented during the review period. The prevalence of gastroschisis amongst neonatal surgical admissions increased from 6% in 2003 to 15% in 2007. 72% of patients weighed less than 2.5 kg at birth and 64% were premature (<37 weeks gestation). 91% were "outborn" with 71% delivered vaginally. Median maternal age was 22.6 years and 57% of mothers were primiparous. Primary abdominal wall closure was possible in 74% of patients. The overall mortality was 43% with sepsis being the leading cause. Staged closure was associated with a higher mortality than primary closure. CONCLUSION: The prevalence of gastroschisis amongst neonatal surgical admissions appears to be increasing. Most babies were "outborn" resulting in delays in diagnosis and referral for surgical management. Despite the availability of NICU and total parenteral nutrition the mortality remains high. Reduction in mortality will depend upon improvements in antenatal diagnosis, primary care and transportation, as well as a reduction in postsurgical sepsis. PMID- 19288119 TI - [Mayer S. Diamantberger (1864-1944). The first person to describe juvenile chronic arthritis]. AB - No evidence can be found in the medical textbooks from the beginning of the twentieth century that rheumatoid arthritis can also occur in children. Isolated cases were first published in the second half of the nineteenth century. In the Hopital Rothschild in Paris several children with this disease were observed and nursed within a short period of time. M.S. Diamantberger, an assistant in this hospital, was intensively involved with these patients and analyzed all previously published cases. This was the theme of his dissertation which was first made public in 1890 and was published in 1891. Diamantberger proved for the first time that there are three totally different types of course in children and particularly mentioned the form affecting the internal organs and the eyes. Therefore he had described the so-called systemic form 6 years before Still and anticipated the oligarthritis accompanying inflammation of the eye. Diamantberger's work found wide acceptance in the French specialist literature. In 1988 this was reprinted in book form, nearly 100 years after the first publication. Nevertheless, only few clinicians in Germany have taken note of this and in the Anglo-American literature Diamantberger has remained unknown. Even today his name is not to be found neither in standard reference works on rheumatology nor in textbooks on pediatrics and the history of medicine. Another Parisian clinician, A. Chauffard, did not fare any better. He reported in 1896 on a systemic form of chronic polyarthritis in adults, the discovery of which is world-wide accredited to E. Bywaters in 1971. CONCLUSION: Many French authors were victims of the language barrier. PMID- 19288121 TI - Evidence for abnormal tau phosphorylation in early aggressive multiple sclerosis. AB - Although progression in multiple sclerosis is pathologically dominated by neurodegeneration, the underlying mechanism is unknown. Abnormal hyperphosphorylation of tau is implicated in the aetiopathogenesis of some common neurodegenerative disorders. We recently demonstrated the association of insoluble tau with established secondary progressive MS, raising the hypothesis that its accumulation is relevant to disease progression. In order to begin to determine the temporal emergence of abnormal tau with disease progression in MS, we examined tau phosphorylation in cerebral tissue from a rare case of early aggressive MS. We report tau hyperphosphorylation occurring in multiple cell types, with biochemical analysis confirming restriction to the soluble fraction. The absence of sarcosyl-insoluble tau fraction in early disease and its presence in secondary progression raises the possibility that insoluble tau accumulates with disease progression. PMID- 19288122 TI - Management of Wolffian duct tumor recurrence without chemotherapy. AB - BACKGROUND: A female adnexal tumor of probable Wolffian origin (FATWO) is a rare neoplasm which is considered to have a low malignant potential. CASE: In 2006, a 60-year-old patient was admitted to hospital with solid tumor on the vaginal cuff confirmed by abdominal/pelvic CT, 6 years after her first FATWO was diagnosed and hysterectomy, bilateral adnexectomy, and omentectomia were done. No chemotherapy was administrated after her first surgery. Pelvic washings, lysis of adhesions, and excision of tumor mass were performed. Pathologic examination revealed malignant epithelial neoplasm of Wolffian origin with morphologic features similar to those of the patient's prior tumor. Immunohistochemical staining was positive for vimentin and partially low positive for cytokeratin 19. No chemotherapy was administrated. Two years after second surgery, the patient was asymptomatic and her clinical, laboratory, CT, and chest X-ray findings were unremarkable. CONCLUSION: Surgical debulking with hysterectomy and bilateral adnexectomy appears as the most successful initial treatment for FATWOs. PMID- 19288120 TI - Oxidatively modified proteins in Alzheimer's disease (AD), mild cognitive impairment and animal models of AD: role of Abeta in pathogenesis. AB - Oxidative stress has been implicated in the pathogenesis of a number of diseases including Alzheimer's disease (AD). The oxidative stress hypothesis of AD pathogenesis, in part, is based on beta-amyloid peptide (Abeta)-induced oxidative stress in both in vitro and in vivo studies. Oxidative modification of the protein may induce structural changes in a protein that might lead to its functional impairment. A number of oxidatively modified brain proteins were identified using redox proteomics in AD, mild cognitive impairment (MCI) and Abeta models of AD, which support a role of Abeta in the alteration of a number of biochemical and cellular processes such as energy metabolism, protein degradation, synaptic function, neuritic growth, neurotransmission, cellular defense system, long term potentiation involved in formation of memory, etc. All the redox proteomics-identified brain proteins fit well with the appearance of the three histopathological hallmarks of AD, i.e., synapse loss, amyloid plaque formation and neurofibrillary tangle formation and suggest a direct or indirect association of the identified proteins with the pathological and/or biochemical alterations in AD. Further, Abeta models of AD strongly support the notion that oxidative stress induced by Abeta may be a driving force in AD pathogenesis. Studies conducted on arguably the earliest stage of AD, MCI, may elucidate the mechanism(s) leading to AD pathogenesis by identifying early markers of the disease, and to develop therapeutic strategies to slow or prevent the progression of AD. In this review, we summarized our findings of redox proteomics identified oxidatively modified proteins in AD, MCI and AD models. PMID- 19288123 TI - Salvage surgery after induction chemotherapy with paclitaxel/cisplatin and primary radiotherapy for advanced laryngeal and hypopharyngeal carcinomas. AB - Induction chemotherapy followed by primary radiotherapy in responders is considered an alternative to surgery for advanced cancer of the larynx and hypopharynx (LHC). Comparison of therapeutic approaches is challenging and must respect oncological and functional outcome as well as quality of life during and after treatment. One aspect of primary radiochemotherapy is the option of salvage surgery in case of residual tumor. The outcome after salvage surgery following new organ-preserving strategies has to be examined. All patients undergoing induction chemotherapy with paclitaxel and cisplatin followed by radiotherapy from 01/96 to 07/05 were included. Salvage surgery was performed either for local recurrence or suspected persistent nodal disease. Complete tumor removal, perioperative morbidity, and overall survival were analyzed in a retrospective study. 28 out of 134 patients underwent salvage surgery after primary treatment with induction chemotherapy and radiotherapy for advanced LHC. 15 patients had laryngectomy (LE) with neck dissection (ND), while 1 patient had lasersurgical partial laryngeal resection with ND for local recurrences. Twelve patients had salvage ND for suspicion of persistent lymph node metastases. 73% of LE patients had major postoperative problems such as pharyngocutaneous fistulas. In 56% of the cases, tumor removal turned out to be microscopically incomplete. Eight out of 12 patients who underwent salvage ND because of suspicious lymph nodes (66%) were free of vital tumor. When metastatic disease was present in the neck (4/12), recurrences occurred in 75% during postoperative follow-up. Only 2 out of 20 patients undergoing surgery for histologically proven recurrence after radiochemotherapy (10%) are actually tumor-free and alive after a mean observation time of 43.9 months. Salvage surgery for local recurrence is associated with high morbidity and poor oncological and functional outcome. ND for suspicious persistent nodal disease after radiochemotherapy can be an over treatment. In our patients, it was burdened with cervical recurrences and distant metastases in presence of histologically confirmed lymph node metastases. In the light of our results, unfavourable outcome after salvage surgery must be pointed out when initially informing patients about different therapeutic options for advanced LHC. PMID- 19288124 TI - Benign paroxysmal positional vertigo after intense physical activity: a report of nine cases. AB - The aim of this study was to report some clinical cases suggesting a possible correlation between benign paroxysmal positional vertigo (BPPV) and intense physical activity. Out of 430 BPPV cases referred to our out-patients clinic, 9 patients, showing symptoms of BPPV arising after an intense period of physical activity, were selected for this study. The posterior semicircular canal was affected in all the nine patients. The canalith repositioning procedure was successful and eliminated vertigo and nystagmus in all patients. During the follow-up period (12 months) all patients continued with the usual physical activity; four of the nine patients showed a recurrence of the BPPV symptoms after a new intense period of exercises: all were successfully treated by a new single Epley repositioning procedure. BPPV due to intense physical activity is a rare condition (9/430) and it may be caused by repeated vibratory vertical accelerations of a minor degree associated with metabolic variations during strenuous exercise. PMID- 19288125 TI - Differential membrane redistribution of P2X receptor isoforms in response to osmotic and hyperglycemic stress in the rat lens. AB - P2X(1, 2, 3, 4, 6 and 7) are all expressed in a differentiation-dependent manner in the rat lens. However, in the lens outer cortex the subcellular distribution of all P2X isoforms is predominantly associated with a pool of receptors located in cytoplasmic vesicles. Here we investigate whether osmotic and hyperglycemic stress can alter the subcellular distribution of this cytoplasmic pool of P2X receptors. We show that in a discrete zone of the deeper outer cortex an isoform and stimulus-specific shift in the subcellular distribution of P2X receptors occurs from the cytoplasm to defined membrane domains. In response to hypertonic stress P2X(1) and P2X(4) isoforms became more closely associated with the broad sides of fiber cells, while under hypotonic conditions P2X(4) and P2X(6) isoforms associate with the narrow side membranes. No such changes in subcellular distribution were observed for P2X(2,3 and 7) isoforms. Lens cultured in 50 mM glucose exhibited cell swelling in this zone but only P2X(4) associated with narrow side membranes. Our results indicate P2X receptors can be differentially recruited to specific membrane domains of lens fiber cells by osmotic and hyperglycemic stress. Furthermore they suggest the involvement of specific P2X isoforms in the regulation of fiber cell volume and the initiation of diabetic cataract. PMID- 19288126 TI - Effects of knee and ankle muscle fatigue on postural control in the unipedal stance. AB - The aim of this study was to compare the effects of acute muscle fatigue of the ankle and knee musculature on postural control by immediate measures after performing fatiguing tasks (POST condition). One group of subjects (n = 8) performed a fatiguing task by voluntary contractions of the triceps surae (group TRI) and the other (n = 9) performed a fatiguing task by voluntary contractions of the quadriceps femoris (group QUA). Each muscle group was exercised until the loss of maximal voluntary contraction torque reached 50% (isokinetic dynamometer). Posture was assessed by measuring the centre of foot pressure (COP) with a force platform during a test of unipedal quiet standing posture with eyes closed. Initially (in PRE condition), the mean COP velocity was not significantly different between group TRI and group QUA. In POST condition, the mean COP velocity increased more in group QUA than in group TRI. The postural control was more impaired by knee muscle fatigue than by ankle muscle fatigue. PMID- 19288127 TI - Electro-physiological parameters of hepatic radiofrequency ablation--a comparison of an in vitro versus an in vivo porcine liver model. AB - BACKGROUND: Radiofrequency ablation (RFA) is an inherent part of curative treatment within a multimodal therapy concept of malignant liver tumors. The biggest problem is the high rate of local recurrences in tumors with a diameter of more than 3 cm because of the high variability and poor reproducibility of the zone of ablation. No imaging modality facilitates monitoring during neither intraoperativ nor percutaneous RFA. This experimental study describes and compares an in vitro and in vivo porcine model by its electro-physiological parameters with the aim of monitoring RFA procedures. MATERIALS AND METHODS: RFA was performed in a perfused in vitro porcine (one RFA per liver) and in vivo porcine model (24 animals) with three different RFA systems (Rita XL 5 cm, Rita XLi 7 cm, LeVeen 5 cm). In the in vivo model, percutaneous placement of the RFA device was guided by native and contrast-enhanced CT scan. The electro-physical parameters during RFA were online (in real time) recorded by a dedicated software. After the RFA, the livers were explanted, sliced, and measured according to the consensus technique. RESULTS: The delivered energy was in vivo versus in vitro: Rita XL 238 +/- 135 kJ versus 135 +/- 53 kJ (p = 0.247); Rita XLi 711 +/- 180 kJ versus 159 +/- 54 (p = 0.016) and with LeVeen 212 +/- 71 kJ (in vivo). The LeVeen system was inconsistent in the in vitro model. This correlates to an energy consumption per ml of necrosis in vivo versus in vitro Rita XL of 8 +/- 3 kJ/ml versus 6.4 +/- 3.9 kJ/ml (p = 0.537), Rita XLi of 10 +/- 6 kJ/ml versus 1.8 +/- 0.2 kJ/ml (p = 0.016), and LeVeen of 14.0 +/- 12 kJ/ml (in vivo). The volume of ablation was in vivo versus in vitro Rita XL 30 +/- 10 ml versus 26 +/- 17 ml (p = 0.329), Rita XLi 90 +/- 58 ml versus 88 +/- 21 ml (p = 0.905), and LeVeen 22 +/- 11 ml versus 50 +/- 12 ml (p = 0.04). The impedance during RFA were in vivo versus in vitro Rita XL 39 +/- 4 Omega versus 50 +/- 14 Omega (p < 0.247), Rita XLi 33 +/- 5 Omega versus 61 +/- 16 Omega (p = 0.016) and LeVeen 31 +/- 2 Omega (in vivo). CONCLUSION: The volume of ablation showed analogue data in vivo and in vitro. The delivered energy and energy consumption was in vivo up to five times (Rita XLi) higher than in vitro and the impedance in vivo was always lower than in vitro. These differences observed between in vivo and in vitro were more pronounced than previously described. Thus the use of an in vitro model for research of the RFA technique must be challenged. The large deployment of the Rita XLi was a problem for percutaneous positioning of the device without direct contact to liver surface or major vessels in 80-kg pigs and to a lesser extent in in vitro liver originating from 130- to 140-kg pigs. Modern RFA systems which generate large volume of tissue necrosis can therefore only be adequately tested in a porcine model with a liver weight of at least 1.5-2 kg. Alternatively, a bovine liver model (with a liver weight up to 10 kg) should be developed in the future. PMID- 19288128 TI - Infant and perinatal pulmonary hypoplasia frequently associated with brainstem hypodevelopment. AB - Recent anatomo-pathological studies have revealed a frequent associated hypoplasia of both arcuate nucleus and lungs in stillbirths. The purpose of this study is to analyze the lung and brainstem development in sudden unexplained perinatal death and sudden infant death syndrome (SIDS). A total of 51 cases were investigated. A complete autopsy was performed in each case. Anatomo-pathologic examination of the central autonomic nervous system included an in-depth study on histological serial sections of the brains where the main structures participating in control of the vital functions are located. The stage of lung development was evaluated by macroscopic and microscopic criteria. In 52.9% of cases, a pulmonary hypoplasia was detected. The pulmonary hypoplasia was significantly more frequent in the SIDS group compared to the sudden perinatal unexplained death groups (p < 0.05). In 72.5% of cases, histological examination of the brainstem on serial sections showed hypodevelopment of the brainstem nuclei, particularly hypoplasia, of the arcuate nucleus (60.8%). In 47.1% of cases, pulmonary hypoplasia was associated with brainstem hypodevelopment. PMID- 19288129 TI - The impact of microRNAs on colorectal cancer. AB - MicroRNAs are small RNAs that regulate gene expression at the post transcriptional level. After their discovery 15 years ago, a new layer of gene regulation was introduced into every field of human biology and medicine. Considering the strong association between genetic alterations and neoplastic diseases, it is not surprising that there is a special focus on miRNAs and cancer. A multitude of experimental studies on colorectal cancer, the most common cancer site and furthermore the second most common cause of death due to cancer, deliver insight into miRNA-mediated, regulatory links to well-known oncogenic and tumour suppressor signalling pathways. Furthermore, several investigations have described the ability of microRNA expression patterns to predict prognosis in colon cancer and support diagnosis of poorly differentiated tumours. In this short review, we give a comprehensive overview focussed on miRNAs in colorectal cancer research. PMID- 19288130 TI - Involvement of Fc(epsilon)R1beta gene polymorphisms in susceptibility to atopy in Korean children with asthma. AB - INTRODUCTION: IgE-dependent activation of mast cells and basophils through the high-affinity IgE receptor (Fc(epsilon)R1) is involved in the pathogenesis of allergen-induced immune responsiveness in atopic disease including bronchial asthma. MATERIALS AND METHODS: We genotyped 650 children for allelic determinants at two polymorphic sites, -109T/C and E237G, in the Fc(epsilon)R1beta gene by SNP IT assays using the SNP stream 25K system. RESULTS: Distributions of the genotype and allele frequencies of Fc(epsilon)R1beta -109T/C and E237G polymorphisms were significantly associated with atopy (P < 0.05) and elevated serum IgE levels. However, differences in the E237G polymorphism did not reach statistical significance after adjustment for multiple comparisons. The genotypes TC or CC at -109T/C were associated with decreased forced expiratory flow(25-75%) in children with asthma (P < 0.05), but this did not reach statistical significance after correction for multiple comparisons. In addition, haplotype 1 (T-A) was associated with atopy susceptibility (P = 0.0069). Analysis of genotype distributions of haplotypes demonstrated a significantly lower PC(20) for homozygous -/- diploids compared with homozygous Ht1/Ht1 (P = 0.0261). CONCLUSION: Polymorphisms in the Fc(epsilon)R1beta gene confer susceptibility to atopy in Korean children and may have a disease-modifying effect on airways of asthmatic patients. PMID- 19288131 TI - Highly sensitive detection of melanoma based on serum proteomic profiling. AB - PURPOSE: There is no available tumor marker that can detect primary melanoma. Proteomics analysis has been proposed as a novel tool that would lead to the discovery of potential new tumor markers. METHODS: We developed a serum proteomic fingerprinting approach coupled with a classification method to determine whether proteomic profiling could discriminate between melanoma and healthy volunteers. A total of 108 serum samples from 30 early-stage [American Joint Committee on Cancer (AJCC) stage I or II] and 30 advanced-stage (AJCC stage III or IV) melanoma patients and 48 healthy volunteers were analyzed by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) utilizing protein chip technology and artificial neural networks. RESULTS: In a first step, a multiprotein classifier was built using a training set of 30 pathologically confirmed melanoma and 24 healthy volunteer serum samples, resulting in good classification accuracy for correct diagnosis and stage classification assignment. Subsequently, our multiprotein classifier was tested in an independent validation set of 30 melanoma and 24 non-cancer serum samples patients, maintained in a good diagnostic accuracy of 98.1% (sensitivity 96.7%, specificity 100%), and 100% stage I/II classification assignment. CONCLUSIONS: Although results remain to be confirmed in larger collective patient cohorts, we could demonstrate the usefulness of proteomic profiling as a sensitive and specific assay to detect melanoma, including non-metastatic melanoma, from the serum. PMID- 19288132 TI - Evaluation of protective effect of multi-epitope DNA vaccine encoding six antigen segments of Toxoplasma gondii in mice. AB - To investigate the vaccine potential of multi-epitope vaccines against toxoplasmosis, a multi-epitope DNA vaccine, eukaryotic plasmid pcDNA3.1/T-ME expressing six antigen segments (SAG1(238-256), SAG1(281-320), GRA1(170-193), GRA4(331-345), GRA4(229-245), and GRA2(171-185)) of Toxoplasma gondii was constructed. We investigated the efficacy of pcDNA3.1/T-ME with or without co administration of a CpG-oligodeoxynucleotide (CpG-ODN) as an adjuvant to protect mice (BALB/c and C57BL/6) against toxoplasmosis. High survival rates were observed in mice immunized with pcDNA3.1/T-ME when challenged with T. gondii RH strain. Lymphocyte proliferation assays, cytokine, and antibody determinations show that mice immunized with pcDNA3.1/T-ME produced stronger humoral and Th1 type cellular immune responses compared to untreated mice or those immunized with empty plasmids. However, co-immunization with CpG-ODN resulted in impaired immune responses. Our data demonstrates that multi-epitope DNA vaccination is a potential strategy for the control of toxoplasmosis and paves the way for further investigations into producing a multi-epitope anti-T. gondii DNA vaccine. PMID- 19288133 TI - Rapid identification of Giardia duodenalis by loop-mediated isothermal amplification (LAMP) from faecal and environmental samples and comparative findings by PCR and real-time PCR methods. AB - The detection of two human-pathogenic Giardia duodenalis assemblages A and B in faecal and water samples by loop-mediated isothermal amplification (LAMP) has been evaluated. The LAMP reaction is reproducible, rapid and specific for the detection G. duodenalis and has lower costs compared to the other molecular assays. This is the first application of LAMP for Giardia detection. PMID- 19288134 TI - Development of a real-time PCR for the differentiation of the G1 and G2/G3 genotypes of Echinococcus granulosus. AB - The present study was aimed at developing a SYBR Green-based real-time polymerase chain reaction (PCR) assay for a rapid differentiation of the genotype G1 from the cluster of genotypes G2/G3 of Echinococcus granulosus, using as marker the 12S mtDNA gene. Eleven hydatid cysts from water buffaloes and 19 from cattle were used. Fourteen samples (identified as G1 using sequencing) showed a mean melting temperature (T (m)) of 76.4 degrees C and 16 samples (identified as G2/G3 using sequencing) showed a mean T (m) of 77.0 degrees C. The detected mean difference of the T (m) of 0.6 degrees C between G1 and G2/G3 genotypes might allow a fast and simple discrimination of these genotypes. In conclusion, the real-time PCR developed in the present study provides a powerful tool for molecular studies on E. granulosus with possibilities for extension to other genotypes using different molecular targets. PMID- 19288135 TI - The interaction between an introduced fish host and local parasite fauna: Neogobius kessleri in the middle Danube River. AB - Parasite communities of introduced fish Neogobius kessleri Gunther (Gobiidae) were studied at five localities in the Slovak section of the Danube River during 2002-2005. Thirty-three metazoan parasite species were identified. All fish were infected with at least two parasite species; most of the parasite species were generalists. At all sampling sites, high susceptibility to local parasites was observed. The parasite community was dominated by three parasite species: glochidia of Anodonta anatina, larval or subadult acanthocephalan Pomphorhynchus laevis, and larval nematode Raphidascaris acus. The infection of both A. anatina and P. laevis was affected by season and habitat type, with higher abundance in spring and more frequent occurrence of A. anatina in side channels and P. laevis in main river channels. At both the component and infracommunity levels, a more diverse parasite community was found in side channels. This habitat was dominated by actively transmitted parasites, whilst endoparasites were more abundant in fish from the main river channel. Larval stages of parasites dominated the endoparasite community at all sampling sites. The introduced N. kessleri was used as intermediate host for most of the recorded parasites, in some cases also as a paratenic host. Finally, the importance of gobies as suitable hosts for local non native parasite species (Anguillicoloides crassus, Anodonta woodina, Hydrozetes lacustris) is discussed. PMID- 19288136 TI - The role of Rubisco and cell walls in the interspecific variation in photosynthetic capacity. AB - Photosynthetic capacity is known to vary considerably among species. Its physiological cause and ecological significance have been one of the most fundamental questions in plant ecophysiology. We studied the contents of Rubisco (a key enzyme of photosynthesis) and cell walls in leaves of 26 species with a large variation in photosynthetic rates. We focused on photosynthetic nitrogen use efficiency (PNUE, photosynthetic rate per nitrogen), which can be expressed as the product of Rubisco-use efficiency (RBUE, photosynthetic rate per Rubisco) and Rubisco nitrogen fraction (RNF, Rubisco nitrogen per total leaf nitrogen). RBUE accounted for 70% of the interspecific variation in PNUE. The variation in RBUE was ascribed partly to stomatal conductance, and other factors such as mesophyll conductance and Rubisco kinetics might also be involved. RNF was also significantly related to PNUE but the correlation was relatively weak. Cell wall nitrogen fraction (WNF, cell wall nitrogen per total leaf nitrogen) increased with increasing leaf mass per area, but there was no correlation between RNF and WNF. These results suggest that nitrogen allocation to cell walls does not explain the variation in PNUE. The difference in PNUE was not caused by a sole factor that was markedly different among species but by several factors each of which was slightly disadvantageous in low PNUE species. PMID- 19288137 TI - Short- and long-term consequences of individual and territory quality in a long lived bird. AB - Site-quality is a major determinant of fitness but its effect can be confounded by individual quality, a relationship that has been little studied in large, long lived vertebrates. The fitness effects of quality estimates depend on the assumption of co-variation between individual and territory quality and can be framed as five working hypotheses: no effect on fitness, exclusive effect of individual quality, exclusive effect of site quality, and independent or interactive effects of the two. We explored such a framework using a medium-sized raptor, the black kite Milvus migrans, as a model species. Individual and territory quality co-varied, but the strength of the relationship varied across different estimates of individual quality (age, body size, or mass residuals). Short-term production of fledglings was related to the independent effects of both individual and territory quality. However, longer-term production of recruits was related solely to territory quality. The disappearance of individual quality effects over the long-term may be caused by antagonistic selective pressures acting during different stages of the life cycle. Our results contribute to a growing appreciation of the long-term fitness-benefits of advantages experienced in early life and highlight the importance of a long-term perspective in studies assessing the effects of individual and territory quality. In our case study, prioritizing sites for conservation on the basis of territory quality may be a feasible pathway to maintain the viability of the population. However, scenarios where such a method could be inefficient have been previously reported, suggesting caution in its application. More studies are needed to understand the generality of the efficiency of priority-setting approaches based on site quality. PMID- 19288138 TI - Production and characterization of lipopeptide biosurfactant by a sponge associated marine actinomycetes Nocardiopsis alba MSA10. AB - A sponge-associated marine actinomycetes Nocardiopsis alba MSA10 was screened and evaluated for the production of biosurfactant. Biosurfactant production was confirmed by conventional screening methods including hemolytic activity, drop collapsing test, oil displacement method, lipase production and emulsification index. The active compound was extracted with three solvents including ethyl acetate, diethyl ether and dichloromethane. The diethyl ether extract was fractionated by TLC and semi-preparative HPLC to isolate the pure compound. In TLC, a single discrete spot was obtained with the R (f) 0.60 and it was extrapolated as valine. Based on the chemical characterization, the active compound was partially confirmed as lipopeptide. The optimum production was attained at pH 7, temperature 30 degrees C, and 1% salinity with glucose and peptone supplementation as carbon and nitrogen sources, respectively. Considering the biosurfactant production potential of N. alba, the strain could be developed for large-scale production of lipopeptide biosurfactant. PMID- 19288139 TI - Change in C1q deposition in C1q nephropathy. PMID- 19288140 TI - Is continuous antibiotic prophylaxis in children with vesicoureteral reflux really ineffective in preventing pyelonephritis? PMID- 19288142 TI - Taking a hard look at the pathogenesis of childhood HIV-associated nephropathy. AB - Childhood human immunodeficiency virus-associated nephropathy (HIVAN) is defined by the presence of proteinuria associated with mesangial hyperplasia and/or global-focal segmental glomerulosclerosis, in combination with the microcystic transformation of renal tubules. This review discusses the pathogenesis of childhood HIVAN and explores how the current pathological paradigm for HIVAN in adults can be applied to children. The Human Immunodeficiency Virus-1 (HIV-1) induces renal epithelial injury in African American children with a genetic susceptibility to develop HIVAN. The mechanism is not well understood, since renal epithelial cells harvested from children with HIVAN do not appear to be productively infected. Children with HIVAN show a renal up-regulation of heparan sulphate proteoglycans and a recruitment of circulating heparin-binding growth factors, chemokines, and mononuclear cells. Macrophages appear to establish a renal HIV-reservoir and transfer viral particles to renal epithelial cells. All of these changes seem to trigger an aberrant and persistent renal epithelial proliferative response. The paradigm that viral products produced by infected renal epithelial cells per se induce the proliferation of these cells is not supported by data available in children with HIVAN. More research is needed to elucidate how HIV-1 induces renal epithelial injury and proliferation in HIV infected children. PMID- 19288143 TI - Imaging anatomy and variation of vertebral artery and bone structure at craniocervical junction. AB - The objective of this article is to display the vertebral artery and bone structure at the craniocervical junction (CJVA and C(0-1-2)) with three dimensional CT angiography (3DCTA) and identify their anatomic features and variations. Eighty-eight subjects without pathology of vertebral artery (VA) and C(0-1-2) were selected from head-neck CTA examination. 3D images were formed with volume rendering (VR) and multiplanar reconstruction (MPR). On the 3D images, CJVA and C(0-1-2) were measured, and their variations were observed. CJVA goes along C(0-1-2) with five curves, of which three curves are visibly away from C(0 1-2), one is 0.0-8.3 mm away at the second curve with 0.0-11.2 mm in width, another is 0.0-9.2 mm away at the fourth with 2.8-14.8 mm and the other is 0.0 6.2 mm away at the fifth. Statistical comparisons show that there is no significant difference in the measurements between left and right, and that the curves become smaller and farther away from C(0-1-2) with the increase of age. CJVA is not equal in size, with the biggest in the fourth curve and the smallest in the fifth. Statistical comparison shows the left CJVA is larger than the right in the fifth curve. Variations were found on CJVA in 16 cases and on C(1) in 12 cases. The anatomy and variations of CJVA and C(0-1-2) are complicated. It is of vital significance to identify their anatomic features in clinical practice. PMID- 19288144 TI - Exploring metallodrug-protein interactions by mass spectrometry: comparisons between platinum coordination complexes and an organometallic ruthenium compound. AB - Electrospray ionisation mass spectrometry was used to analyse the reactions of metal compounds with mixtures of selected proteins. Three representative medicinally relevant compounds, cisplatin, transplatin and the organometallic ruthenium compound RAPTA-C, were reacted with a pool of three proteins, ubiquitin, cytochrome c and superoxide dismutase, and the reaction products were analysed using high-resolution mass spectrometry. Highly informative electrospray ionisation mass spectra were acquired following careful optimisation of the experimental conditions. The formation of metal-protein adducts was clearly observed for the three proteins. In addition, valuable information was obtained on the nature of the protein-bound metallofragments, on their distribution among the three different proteins and on the binding kinetics. The platinum compounds were less reactive and considerably less selective in protein binding than RAPTA C, which showed a high affinity towards ubiquitin and cytochrome c, but not superoxide dismutase. In addition, competition studies between cisplatin and RAPTA-C showed that the two metallodrugs have affinities for the same amino acid residues on protein binding. PMID- 19288145 TI - Linking conformation change to hemoglobin activation via chain-selective time resolved resonance Raman spectroscopy of protoheme/mesoheme hybrids. AB - Time-resolved resonance Raman (RR) spectra are reported for hemoglobin (Hb) tetramers, in which the alpha and beta chains are selectively substituted with mesoheme. The Soret absorption band shift in mesoheme relative to protoheme permits chain-selective recording of heme RR spectra. The evolution of these spectra following HbCO photolysis shows that the geminate recombination rates and the yields are the same for the two chains, consistent with recent results on (15)N-heme isotopomer hybrids. The spectra also reveal systematic shifts in the deoxyheme nu (4) and nu (Fe-His) RR bands, which are anticorrelated. These shifts are resolved for the successive intermediates in the protein structure, which have previously been determined from time-resolved UV RR spectra. Both chains show Fe-His bond compression in the immediate photoproduct, which relaxes during the formation of the first intermediate, R(deoxy) (0.07 micros), in which the proximal F-helix is proposed to move away from the heme. Subsequently, the Fe-His bond weakens, more so for the alpha chains than for the beta chains. The weakening is gradual for the beta chains, but is abrupt for the alpha chains, coinciding with completion of the R-T quaternary transition, at 20 micros. Since the transition from fast- to slow-rebinding Hb also occurs at 20 micros, the drop in the alpha chain nu (Fe-His) supports the localization of ligation restraint to tension in the Fe-His bond, at least in the alpha chains. The mechanism is more complex in the beta chains. PMID- 19288146 TI - Parameterization of aromatic azido groups: application as photoaffinity probes in molecular dynamics studies. AB - The accuracy of molecular dynamics (MD) simulations is limited by the availability of parameters for the molecular system of interest. In most force fields, parameters of common chemical groups are already present. With the development of novel small organic molecules as probes to study biological systems, more chemical groups require parameterization. An azide group is often used in studies of biological systems but computational studies are still impeded by the lack of parameters. In this paper, we present a set of molecular mechanics (MM) parameters for aromatic and aliphatic azido groups, and their application in MD simulations of a photoaffinity probe currently used in our laboratory for mapping binding modes available in the active site of histone deacetylases. The parameters were developed for the generalized Amber force field (GAFF) using density functional theory (DFT) calculations at B3LYP 6-311G(d) level. The parameters were validated by geometry optimization and MD simulations. PMID- 19288148 TI - Laryngocele and epiglottic cyst as rare causes of obstructive sleep apnea. AB - INTRODUCTION: Upper airway pathologies have been reported to cause obstructive sleep apnea (OSA). CASE REPORT: We present two rare cases of OSA, one with laryngocele secondary to partial laryngectomy and the other with epiglottic cyst. CONCLUSION: In the management of cases with OSA symptoms, a detailed evaluation of upper airways, including endoscopic and radiological examinations as well as polysomnography, should be done. PMID- 19288149 TI - Effect of a low dose of sea buckthorn berries on circulating concentrations of cholesterol, triacylglycerols, and flavonols in healthy adults. AB - BACKGROUND: Epidemiological studies indicate beneficial effects of flavonoids on cardiovascular disease (CVD) risk. AIM OF THE STUDY: To study the effect of flavonoid-rich sea buckthorn berry (SBB) on circulating lipid markers associated with CVD risk and plasma flavonol concentration. Also investigated was whether changes in the circulating flavonol concentrations correlate with the SBB induced changes in C-reactive protein (CRP) concentration observed previously. SUBJECTS AND METHODS: In all 229 healthy participants completed the randomized double blind study and consumed daily 28 g of SBB or placebo for 3 months. Fasting blood samples for the analysis of lipid markers and flavonols were obtained at the beginning and end of the study. RESULTS: Compared to the placebo, the consumption of SBB increased the plasma concentration of the flavonols quercetin and isorhamnetin significantly [treatment differences 3.0 ng/ml (P = 0.03) and 3.9 ng/ml (P < 0.01), respectively]. The increase of kaempferol concentration was not significant [treatment difference 0.7 ng/ml (P = 0.08)]. SBB did not affect the serum total, HDL, LDL cholesterol, or the serum triacylglycerol concentrations. There was no correlation between the changes in flavonol and CRP concentrations of participants. CONCLUSIONS: The consumption of SBB significantly increased the fasting plasma concentration of quercetin and isorhamnetin indicating that it is a good dietary source of flavonols. However, this did not convert to affecting the circulating concentrations of lipid markers in healthy, normolipidemic adults having healthy diets. PMID- 19288150 TI - Gastric cancer in relation to the intake of nutrients involved in one-carbon metabolism among MTHFR 677 TT carriers. AB - BACKGROUND: DNA methylation is an important epigenetic process for transcriptional control of human genome including those genes involved in cancer initiation and progression. Clinical studies have suggested that biological explanation to the protective effect of some nutrients could be linked with the DNA methylation. Folate is a primary methyl donor nutrient; it has been shown to play a key role in DNA methylation, repair and synthesis, by acting as co-factors and/or substrates in this metabolic pathway. Likewise, activity of a key enzyme, the methylenetetrahydrofolate reductase (MTHFR) has also been shown to influence DNA methylation. Overall, these findings support the notion that dietary intake as well as genetic factors play a role in one-carbon metabolism. AIM OF THE STUDY: This study is to evaluate the dietary intake of nutrients involved in one carbon metabolism and the genotype of MTHFR 677 C > T with respect to GC risk. METHODS: We carried out in January 2004 a population-based case-control study in the metropolitan area of Mexico City. A total of 248 histological confirmed GC patients were recruited from nine tertiary hospitals, along with 478 age and sex matched controls. Nutrient intake was estimated from food frequency questionnaire; the MTHFR 677C > T genotype was determined by PCR-RFLP analysis. RESULTS: A significant reduction in diffuse GC risk was observed for MTHFR 677 TT genotype among individuals with high consumption of folate (OR = 0.23; 95% CI 0.06-0.84), choline (OR = 0.55; 95% CI 0.33-0.9) and Vitamin B(6) (OR = 0.59; 95% CI 0.36-0.96) compared to MTHFR 677 CC + CT carriers. Among subjects with low consumption of methionine, a reduced risk of diffuse GC was also detected (OR = 0.40; 95% CI 0.16-0.97). In contrast, carriers of the MTHFR 677 TT genotype with a low consumption of folate had a significant increased risk of intestinal GC (OR = 1.88 95% CI 1.02-3.47). A folate-MTHFR 677 C > T interaction in the borderline of significance (P = 0.055) was detected. CONCLUSIONS: It is probable that GC prevention requires dietary recommendations according to the individual genotype; nevertheless, the available information to this respect is still very limited. PMID- 19288151 TI - Regional, age-dependent, and genotype-dependent differences in ventricular action potential duration and activation time in 410 Langendorff-perfused mouse hearts. AB - Although numerous studies have reported the effects of genetic alterations on murine electrophysiology, the range of normal values for ventricular activation, repolarization, and arrhythmias in mouse hearts is not known. We analyzed right ventricular (RV), left ventricular (LV), and septal activation times, monophasic action potential durations (APD), and right ventricular effective refractory periods during spontaneous rhythm, induced AV nodal block, right ventricular pacing (100-300 ms paced cycle length), and programmed stimulation in 410 beating, Langendorff-perfused, wild-type mouse hearts of CD1, DBAC3H, FVBN, C57/Bl6, and hybrid backgrounds (age 203 +/- 132 days). Action potential duration was longer at longer cycle lengths. LV-APD prolonged more than RV-APD, resulting in an increased heterogeneity of APD at longer pacing cycle lengths. Higher heart weight/body weight ratio and DBAC3H and FVB/N backgrounds were associated with long APD, C57Bl/6 background was associated with short APD. Activation times were longer in older hearts. There were no clear-cut sex-dependent APD differences. Sustained spontaneous arrhythmias occurred in 1% of hearts, non-sustained arrhythmias in 18%. Induction of AV block and C57Bl/6 genetic background were associated with spontaneous arrhythmias. Programmed stimulation induced arrhythmias in 51% of hearts. Inducible arrhythmias were associated with advanced age and shorter refractory periods. Ventricular APD in beating mouse hearts show rate- and site-dependent changes comparable to man and large animals. Bradycardia provokes spontaneous arrhythmias in mouse heart, while age-dependent conduction slowing and short refractory periods predispose to induced arrhythmias. Genetic background influences repolarization and arrhythmogenesis. These findings provide systematic data for the design and interpretation of arrhythmia studies in murine disease models. PMID- 19288152 TI - Effect of insulin and glucose infusion on myocardial infarction size in uraemic rats. AB - The post myocardial infarction (MI) mortality rate is high in renal patients. One possible explanation is the reduced ischemia tolerance caused by uraemia. Previous investigations showed larger MI size in uraemic rats when compared with sham-operated controls. To explore a possible link between uraemic insulin resistance syndrome and MI size in uraemia, we studied an intervention model with administration of insulin and glucose during acute MI in subtotally nephrectomized (SNX) rats and sham-operated controls. In 16 SNX rats and 16 sham operated controls, the left coronary artery was ligated for 60 min, followed by reperfusion for 90 min. To visualize the perfused myocardium, lissamine-green ink was injected. The nonperfused area (lissamine exclusion) and the area of total infarction (TTC stain) were assessed in sections of the left ventricle (LV) using image analysis. While eight SNX rats and eight sham-operated controls were treated with a placebo during the procedure, the other animals received an insulin bolus of 85 mU/kg and then a continuous insulin infusion of 8 mU/kg per minute. Blood glucose levels were clamped to baseline levels with an infusion of 25% glucose. Insulin receptor substrates (IRS-1 and IRS-2) and glucose transporter (GLUT 4) were studied by western blot in another seven SNX and seven sham-operated controls without further intervention. The infarcted area, given as a proportion of the nonperfused risk area, was not different in sham-operated controls treated with a hyperinsulinaemic clamp versus untreated (0.55 +/- 0.07 vs. 0.51 +/- 0.13, p = 0.477). The eight SNX animals treated with the hyperinsulinaemic clamp utilized significantly less glucose to stabilize baseline glucose levels when compared with the sham-operated controls (5,637 vs. 3,207 microl Glc 25%, p = 0.007). The infarcted area was significantly lower in SNX rats treated with the hyperinsulinaemic clamp compared to non-treated SNX animals (0.56 +/- 0.06 vs. 0.79 +/- 0.09, p < 0.001). SNX rats with the insulin clamp had the same infarcted area size as sham-operated controls (0.56 +/- 0.06 vs. 0.51 +/ 0.13, p = 0.357). Western blotting did not show any change in the expression of GLUT 4 and IRS-1/IRS-2 in SNX animals when compared with sham-operated controls. The size of MI in uraemic rats is significantly reduced by a glucose/insulin infusion. The results suggest an insulin resistance in uraemic rats with similar benefits of glucose/insulin application during acute MI, as found in diabetic individuals. Further analysis did not reveal a down regulation in GLUT 4 and IRS 1/IRS-2. PMID- 19288153 TI - Time course and mechanisms of left ventricular systolic and diastolic dysfunction in monocrotaline-induced pulmonary hypertension. AB - Although pulmonary hypertension (PH) selectively overloads the right ventricle (RV), neuroendocrine activation and intrinsic myocardial dysfunction have been described in the left ventricle (LV). In order to establish the timing of LV dysfunction development in PH and to clarify underlying molecular changes, Wistar rats were studied 4 and 6 weeks after subcutaneous injection of monocrotaline (MCT) 60 mg/kg (MCT-4, n = 11; MCT-6, n = 11) or vehicle (Ctrl-4, n = 11; Ctrl-6, n = 11). Acute single beat stepwise increases of systolic pressure were performed from baseline to isovolumetric (LVPiso). This hemodynamic stress was used to detect early changes in LV performance. Neurohumoral activation was evaluated by measuring angiotensin-converting enzyme (ACE) and endothelin-1 (ET-1) LV mRNA levels. Cardiomyocyte apoptosis was evaluated by TUNEL assay. Extracellular matrix composition was evaluated by tenascin-C mRNA levels and interstitial collagen content. Myosin heavy chain (MHC) composition of the LV was studied by protein quantification. MCT treatment increased RV pressures and RV/LV weight ratio, without changing LV end-diastolic pressures or dimensions. Baseline LV dysfunction were present only in MCT-6 rats. Afterload elevations prolonged tau and upward-shifted end-diastolic pressure dimension relations in MCT-4 and even more in MCT-6. MHC-isoform switch, ACE upregulation and cardiomyocyte apoptosis were present in both MCT groups. Rats with severe PH develop LV dysfunction associated with ET-1 and tenascin-C overexpression. Diastolic dysfunction, however, could be elicited at earlier stages in response to hemodynamic stress, when only LV molecular changes, such as MHC isoform switch, ACE upregulation, and myocardial apoptosis were present. PMID- 19288154 TI - Evaluation of angiographic and technical aspects of carotid stenting with diffusion-weighted magnetic resonance imaging. AB - The detection of clinically silent ischemic lesions on postprocedural diffusion weighted magnetic resonance images has become a preferred method for the description of embolic risks. The purpose of this single-center study was to evaluate whether diffusion-weighted imaging (DWI) could determine material related or technical risk factors of filter-protected carotid stenting. Eighty four patients with symptomatic severe (> or = 60%) carotid artery stenoses received filter-protected carotid stenting. Standard DWI (b = 1000) was performed within 48 h before and after carotid stenting. The occurrence and load of new postinterventional DWI lesions were assessed. Multivariate analysis was performed to determine risk factors associated with DWI lesions, with emphasis on technical factors such as use of different access devices (guiding catheter method vs. long carotid sheath method), type of stent (open-cell nitinol stent vs. closed-cell Wallstent), and protective device (filters with 80-microm vs. 110-120-microm pore size). Markers for generalized atherosclerosis and for degree and site of stenosis were assessed to allow comparison of adequate risk profiles. Access, protective device, and stent type were not significantly associated with new embolic DWI lesions when we compared patients with equivalent risk profiles (long carotid sheath method 48% [11 of 23] vs. guiding catheter method 44% [27 of 61], Wallstent 47% [15 of 32] vs. nitinol stent 44% [23 of 52], and small pore size filter 61% [11 of 18] vs. large pore size filter 41% [27 of 66]). Single-center DWI studies with a moderate number of cases are inadequate for proper assessment of the embolic risk of technical- or material-related risk factors in carotid stenting. Larger multicenter studies with more cases are needed. PMID- 19288155 TI - Protein profile of Nomuraea rileyi spore isolated from infected silkworm. AB - Nomuraea rileyi (N. rileyi) is the causative agent of the silkworm, Bombyx mori, green muscardine which can cause severe worldwide economical loss in sericulture. Little is known about N. rileyi at the protein level for this entomopathogenic parasite which belongs to the Ascomycota. Here, we employed proteomic-based approach to identify proteins of N. rileyi spores collected from the dead silkworm. In all, 252 proteins were separated by two-dimensional gel electrophoresis (2-DE), and were subjected to mass spectrometry (MS) analysis, 121 proteins have good MS signal, and 24 of them were identified due to unavailability of genomic information from N. rileyi. This data will be helpful in understanding the biochemistry of N. rileyi. PMID- 19288156 TI - A high-resolution, three-dimensional thin endoscope for fetal surgery. AB - BACKGROUND: Fetal surgery is receiving considerable attention. However, surgeons must have great skill to perform this surgery. For assisting with the operation, the three-dimensional (3D) endoscope is very useful because it allows the surgeon depth perception. However, the diameter of existing 3D endoscopes is approximately 10 mm. Therefore, the authors have developed a high-resolution, thin, 3D endoscope for use in fetal surgery. METHODS: The authors' system uses two 1/10-in. micro charge-coupled device (CCD) cameras at the tip of the endoscope and achieves a diameter of 5.4 mm. The endoscope's angle of convergence is 2.6 masculine, which very closely approximates the angle of convergence for humans. Thus, the surgeon experiences little visual fatigue. The view angle is 87 masculine. RESULTS: The authors compared image quality and depth perception between their system and conventional 3D and 2D endoscopes. Theoretical investigation of image quality allowed the surgeon to distinguish a line 0.2 to 0.25 mm wide. Furthermore, the depth perception with the thin 3D endoscope was almost the same as with an 11-mm normal 3D endoscope. In addition, with the 3D endoscope, a higher percentage of questions were answered correctly in the depth perception evaluation experiment in a water environment than with the 2D instrument. CONCLUSION: According to these experiments, the thin 3D endoscope has a sufficiently high image quality and depth perception even in a water environment. PMID- 19288157 TI - Laparoscopic single-site surgery for placement of an adjustable gastric band: initial experience. AB - BACKGROUND: Laparoendoscopic single-site (LESS) surgery for cholecystectomy and appendectomy are described in the literature. The benefits of these procedures compared with traditional laparoscopic approaches have yet to be determined. To date, no series of LESS surgeries for placement of an adjustable gastric band has been published or documented. This study aimed to determine the safety and feasibility of LESS surgery for placement of an adjustable gastric band. METHODS: From December 2007 to June 2008, LESS surgery to place an adjustable gastric band via a transumbilical incision was performed for 10 patients with institutional review board approval. Essentially, multiple ports were placed through a single incision in the umbilicus to allow for liver retraction, visualization, and working instruments. All critical steps using a standard pars flaccida technique were performed without alteration. RESULTS: For this study, 10 patients (9 women and 1 man) were carefully selected. These patients ranged in age from 32 to 61 years (mean, 47 years) and had a mean body mass index (BMI) of 42 kg/m2 (range, 35-45 kg/m2). The patients were selected for absence of both hepatomegaly and central obesity. Superobese patients were not considered for inclusion in the study. The mean operative time was 1 h and 10 min (range, 53 min to 1 h and 48 min). All the patients were discharged home within 23 h of admission, and no perioperative complications were noted. In addition, no wound-related complications occurred. Notably, only 2 of the 10 patients required the use of narcotic analgesia after discharge from the recovery room. There were no intra- or postoperative complications. CONCLUSIONS: In our experience, LESS surgery for adjustable gastric banding shows this technique to be both feasible and safe for selected patients. Although technical limitations exist that will be improved upon, further studies are needed to compare LESS surgery for placement of an adjustable gastric band with traditional laparoscopic techniques. PMID- 19288158 TI - Two-year results of a feasibility study on antireflux transoral incisionless fundoplication using EsophyX. AB - BACKGROUND: A feasibility study (n = 19) evaluated the safety and initial efficacy of transoral incisionless fundoplication (TIF) for the treatment of gastroesophageal disease (GERD). The results at 1 year (n = 17) indicated that TIF was safe and had a significant effect on reducing GERD symptoms, proton pump inhibitor (PPI) usage, acid exposure, and small hiatal hernia. This study was designed to evaluate the long-term safety and durability of TIF. METHODS: Fourteen patients (50% female; median age, 34 years) completed the 2-year follow up assessment tests. Three patients were excluded from the study after 1 year because two of them underwent retreatment and one was lost to follow-up. RESULTS: At 2 years, no adverse events related to TIF were reported. A >or=50% improvement in GERD-HRQL scores compared with those at baseline on PPIs was sustained by 64% of patients. TIF was effective in eliminating heartburn in 93% of patients and daily PPI therapy in 71% of patients. Significantly (p < 0.05) more patients were able to consume reflux-causing foods and maintain lifestyle activities without GERD symptoms compared with baseline on PPIs. Fundoplications were durable and maintained their geometric dimensions. TIF was effective in eliminating hiatal hernia in 60% of patients and esophagitis in 55% of patients. Global assessment of all outcomes in each patient revealed that 79% of patients experienced complete cure (29%) or remission (50%) of GERD at 2 years after TIF. CONCLUSION: The results at 2 years supported the long-term safety and durability of TIF and its sustained effect on the elimination of heartburn, esophagitis, 30, and cancer patients. Heart failure patients with worse heart failure-related health status had a greater number of physical symptoms (13.2 vs. 8.6, p = 0.03), higher depression scores (6.7 vs. 3.2, p = 0.001), and lower spiritual well-being (29.0 vs. 38.9, p < 0.01) than patients with advanced cancer. CONCLUSIONS: Patients with symptomatic heart failure and advanced cancer have similar needs for palliative care as assessed by symptom burden, depression, and spiritual well-being. This implies that heart failure patients, particularly those with more severe heart failure, need the option of palliative care just as cancer patients do. PMID- 19288161 TI - A subset of myeloid dendritic cells derived from peripheral blood monocytes represented a predominant subset characterized by their potential tumor inhibiting activity. AB - Besides their role as potent antigen-presenting cells, myeloid dendritic cells (MDCs), but not plasmacytoid dendritic cells (PDCs), have been reported to have cytotoxic or cytostatic activity on some tumor cells. In this article, we analyzed the tumoristatic potential of a distinct peripheral blood monocyte derived MDC subset which co-expressed PDC-specific marker CD123. CD123(+) MDCs represented a subset of small-sized DCs and accounted for 45-60% of peripheral blood monocytes cultured with granulocyte-macrophage colony-stimulating factor and interleukine-4 (IL-4) for 7 d. They exhibited more significant antiproliferative activity toward hematological tumor cell lines of Jurkat, HL60, and myelodysplastic syndromes over-leukemia than CD123(-) MDCs even at a low effecter/target ratio. Pretreatment of MDC and their supernatant with TRAIL-R2:Fc significantly reduced the tumoristatic effect of CD123(+) MDCs but not of CD123( ) MDCs and their supernatant. CD123(+) MDCs expressed higher level of cytoplasmic TNF-alpha-related apoptosis-inducing ligand (TRAIL) than CD123(-) MDCs, whereas both expressed very little surface and soluble TRAIL. These results reveal that CD123(+) cells represented a predominant subset of MDCs generated from peripheral blood monocytes in vitro, characterized by their potential tumoristic activity partially via cytoplasmic TRAIL. PMID- 19288162 TI - Measurement of the mechanical properties of bone: a recent history. AB - Much progress has been made in the last 50 years in our understanding of bone's mechanical properties, and the reasons it has these properties and not others. The question is to what extent these advances have arisen from an increase in the techniques available for the study of bone, and how much stems from an increased understanding of the basic processes involved. Although considerable enlightenment has come from the transfer of ideas from the physical sciences, in particular materials science, the author argues that most increases have come from the vastly increased power and resolution of the observational and mechanical techniques available. Even so, the remarkably hierarchical nature of bone's structure makes it an almost uniquely difficult material to understand properly, and much remains to be done to marry explanations at the macro-, micro- and nanolevels to obtain a full understanding of bone mechanics. PMID- 19288163 TI - Risk stratification with attenuation corrected stress Tc-99m sestamibi SPECT myocardial perfusion imaging in the absence of ECG-gating due to arrhythmias. AB - BACKGROUND: Despite growing recognition of attenuation correction (AC) utilizing an external radiation source, prognostic studies using AC are lacking. METHODS: Consecutive patients (n = 419) who underwent stress Tc-99m sestamibi SPECT imaging with AC but without ECG-gating, due to arrhythmia, were followed for cardiac death or non-fatal myocardial infarction (MI). A 17-segment model and the summed stress score (SSS) were used to classify both the non-AC and AC images. RESULTS: The 419 patients had a mean age of 71.5 +/- 11.7 years and most (70.6%) underwent pharmacologic stress. In follow-up, 35 (8.4%) patients suffered an adverse cardiac event. Patients with AC-SSS 1-3 and AC-SSS 4-8 had similar cardiac event rates (11.4% vs 10.5%, P = NS). Accordingly, AC-SSS cutoffs of 0, 1 8, and >8 were selected to classify perfusion as normal, mildly abnormal, and moderately to severely abnormal with annualized event rates of 2.1%, 10.8%, and 18.7%, respectively (P < .001). In multivariable analysis, AC-SSS >8 was the most powerful predictor of cardiac events followed by AC-SSS 1-8, history of CAD, age >75 and pharmacologic stress. CONCLUSIONS: AC provides powerful risk stratification when added to clinical variables in patients undergoing stress Tc 99m sestamibi SPECT imaging without ECG-gating. Moreover, smaller/less severe defects on AC data are more significant than if the same defects were seen on non AC data. PMID- 19288164 TI - Mechanism for myocardial localization and rapid liver clearance of Tc-99m-N-MPO: a new perfusion radiotracer for heart imaging. AB - BACKGROUND: [Tc-99m-N(mpo)(PNP5)](+) (Tc-99m-N-MPO: Hmpo = 2-mercaptopyridine N oxide and PNP5 = N-ethoxyethyl-N,N-bis[2-(bis(3 methoxypropyl)phosphino)ethyl]amine) is a new Tc-99m radiotracer useful for myocardial perfusion imaging. The main objective of this study is to elucidate the mechanism for myocardial localization and fast liver clearance of Tc-99m-N MPO in comparison with Tc-99m-sestamibi ([Tc-99m-(MIBI)(6)](+): MIBI = 2-methoxy 2-methylpropylisonitrile). METHODS AND RESULTS: Subcellular distribution of Tc 99m-N-MPO and Tc-99m-sestamibi was examined in the excised Sprague-Dawley (SD) rat myocardium. Biodistribution and planar imaging studies were performed using SD rats in the absence/presence of Cyclosporin-A. Due to negative plasma and mitochondrial potentials, 84.5% +/- 3.2% of Tc-99m-N-MPO was found in the mitochondrial fraction as compared to 88.0% +/- 1.5% of Tc-99m-sestamibi. There was no significant difference in their mitochondrial accumulation. Tc-99m-N-MPO was also able to retain its chemical integrity in rat myocardium. Pre-treatment of SD rats with Cys-A result in significant increase in the kidney and liver uptake of Tc-99m-N-MPO. CONCLUSION: Tc-99m-N-MPO and Tc-99m-sestamibi share almost identical subcellular distribution and localization mechanism. The MDR transport function of hepatocytes and renal cells is responsible for the fast clearance kinetics of Tc-99m-N-MPO from liver and kidneys, respectively. Tc-99m-N MPO is a very promising myocardial perfusion radiotracer with favorable biodistribution properties and rapid liver clearance. PMID- 19288167 TI - Cost variation in child and adolescent psychiatric inpatient treatment. AB - Publicly available costs data for child and adolescent psychiatric inpatient services do not allow links to be made with patients' needs and outcomes. Without this information commissioners may reduce the role of inpatient services on the basis of budgetary impacts alone. This study estimates the support costs before, during and after an inpatient admission and explores the associations between costs, needs and outcomes. A detailed prospective cohort study of eight child and adolescent units was undertaken in which participants were assessed at referral, admission, decision to discharge and 1 year later. Mean admission costs were pound24,120, although the range was wide. Associations were found between costs and patients' global impairment, age and exclusion status. Support costs after admission were similar to pre-admission costs, but there was some evidence to suggest that services were better targeted. Moves in England to develop national tariffs for inpatient psychiatric episodes should be based on the likely cost of the episode of treatment rather than costs per day, and good commissioning requires more information on the predictors of such costs. PMID- 19288168 TI - Effects of maternal and paternal smoking on attentional control in children with and without ADHD. AB - Maternal smoking during pregnancy is a risk factor for attention deficit/hyperactivity disorder (ADHD), but data on its adverse effects on cognitive functioning are sparse and inconsistent. Since the effect of maternal smoking during pregnancy may be due to correlated genetic risk factors rather than being a pure environmental effect, we examined the effect of prenatal exposure to smoking on attentional control, taking into account the effects of both maternal and paternal smoking, and examined whether these effects were genetically mediated by parental genotypes. We further examined whether the effect of prenatal exposure to smoking on attentional control interacted with genotypes of the child. Participants were 79 children with ADHD, ascertained for the International Multi-centre ADHD Gene project (IMAGE), and 105 normal controls. Attentional control was assessed by a visual continuous performance task. Three genetic risk factors for ADHD (DRD4 7-repeat allele of the exon 3 variable number of tandem repeats (VNTR), DAT1 10/10 genotype of the VNTR located in the 3' untranslated region, and the DAT1 6/6 genotype of the intron 8 VNTR) were included in the analyses. Paternal smoking had a negative effect on attentional control in children with ADHD and this effect appeared to be mediated by genetic risk factors. The prenatal smoking effect did not interact with genotypes of the child. Maternal smoking had no main effect on attentional control, which may be due to lower smoking rates. This study suggests that the effects of paternal smoking on attentional control in children with ADHD should be considered a proxy for ADHD and/or smoking risk genes. Future studies should examine if the results can be generalized to other cognitive domains. PMID- 19288169 TI - Report of the Asian Forum of Chronic Kidney Disease Initiative (AFCKDI) 2007. "Current status and perspective of CKD in Asia": diversity and specificity among Asian countries. AB - The Japanese Society of Nephrology (JSN) sponsored the Asian Forum of CKD Initiative (AFCKDI) 2007 with the support of the International Society of Nephrology-Commission for Global Advancement in Nephrology (ISN-COMGAN), Asian Pacific Society of Nephrology (APSN), the Kidney Disease: Improving Global Outcome (KDIGO) and other national societies of nephrology in the Asian Pacific region on 27-28 May 2007 in Hamamatsu City, Japan. An international organising committee was established by leading experts of the CKD initiative. The main objective of this forum was to clarify the current status and perspectives of CKD and to promote coordination, collaboration and integration of initiatives in the Asian Pacific region. The forum received 56 papers from 16 countries; it began with the symposium "A Challenge to CKD in the world" and was followed by the ISN COMGAN affiliated workshop "Current status and perspective of CKD in Asia". The second day was dedicated to discussion on the evaluation, surveillance and intervention in CKD in this area. At the end of the forum, we decided on the future plan as follows: (1) The AFCKDI will provide opportunities annually or biannually for every person who promotes CKD initiatives in the Asian Pacific region to join together and build consensus for action; (2) the second forum will be held in Kuala Lumpur on 4 May 2008 at the time of the 11th Asian Pacific Congress of Nephrology (APCN). Zaki Morad, President of the 11th APCN, will host the second forum; (3) the International Organising Committee (IOC) of the 1st AFCKDI will continue its function by adding other experts, including the organisers of the APCN; (4) the AFCKDI is not an organisation by itself, nor does it belong to any society, but is organised by each host national society of nephrology. The IOC will assist the domestic committee for the success of the forum and will assure the continuation of the mission; (5) in order to organise the forum and promote CKD initiatives in the Asia Pacific region, the AFCKDI will look for support by both national and international societies. The AFCKDI will keep an intimate and mutual relation with the ISN, APSN and KDIGO. PMID- 19288170 TI - Prevalence of joint replacement surgery in rheumatoid arthritis patients: cross sectional analysis in a large observational cohort in Japan. AB - The purpose of this study was to describe the prevalence of total joint arthroplasty (TJA) in Japanese rheumatoid arthritis (RA) patients undergoing conventional drug treatment in a large observational cohort in Japan. A total of 5,177 RA patients were studied for the prevalence of TJA, who were enrolled in the NinJa database during the fiscal year of 2006. The cases of 2,695 RA patients with more than ten years of disease duration were extracted and subjected to further analysis. The prevalence of TJA increased in accordance with the disease duration, and the prevalence was markedly increased after ten years. Among the 2,695 patients with more than ten years of disease duration, 1,431 TJAs were performed in 645 (24.6%) patients. The patients with TJA had higher disease activity than those without TJA. In this cross-sectional study, TJAs were performed in approximately a quarter of the Japanese RA patients with more than ten years of disease duration. The result showed that patients with higher disease activity required TJA. PMID- 19288171 TI - Immediate reduction of white blood cell count after tocilizumab administration was observed in some cases. PMID- 19288172 TI - India's stem cell research and development perspectives. PMID- 19288173 TI - Preventive usage of broad spectrum chemokine inhibitor NR58-3.14.3 reduces the severity of pulmonary and hepatic graft-versus-host disease. AB - Pulmonary graft-versus-host disease (pGVHD) is a major complication after allogeneic bone marrow transplantation (BMT), which involves donor leukocyte migration into the lung along chemokine gradients, leading to pulmonary dysfunction and respiratory insufficiency. As broad spectrum chemokine inhibitor (BSCI) NR58-3.14.3 suppresses leukocyte migration in response to various chemokines, including CCL2, CCL3, CCL5, we investigated the effects of NR58 3.14.3 on the evolution of pGVHD. Lethally irradiated B6D2F1 mice received BMT from syngeneic (B6D2F1) or allogeneic (C57BL/6) donors, and animals were treated with either NR58-3.14.3 or vehicle control from day -1 to day +14. At week 6, in allogeneic recipients that received BSCI, inflammatory cell infiltrates in the lung were decreased, and reduced histopathologic changes translated into improved pulmonary function when compared to allo-controls. Acute GVHD of the liver was also diminished, whereas no differences were seen in the gut. Alloantigen dependent splenic T cell expansion and systemic TNF-alpha and IFN-gamma levels were comparable in NR58-3.14.3-treated animals and allo-controls. No suppressive effect of NR58-3.14.3 on CTL cytotoxicity was found, and diminished cellular infiltrates in lung and liver were most likely due to decreased migration of mononuclear cells. Therefore, novel approaches involving BSCIs may provide a promising tool in the management of pGVHD. PMID- 19288174 TI - Roles of TGFbeta and BMP during valvulo-septal endocardial cushion formation. AB - The primordia of valves and the atrioventricular septum arise from endocardial cushion tissue that is formed in the outflow tract (OFT) and in the atrioventricular (AV) regions during cardiogenesis. Abnormal development of the endocardial cushion results in various congenital heart diseases. Endocardial epithelial-mesenchymal transformation (EMT) is a critical process in cushion tissue formation and is regulated by many factors, such as growth factors, intercellular signaling molecules, transcription factors, and extracellular matrices. A signal that is produced by the myocardium of the AV and OFT regions and transferred to the adjacent endocardium across the extracellular matrix mediates EMT. Studies in vitro and genetic analyses have shown that transforming growth factor beta and bone morphogenetic protein play central roles in the regulation of EMT during cushion tissue formation. PMID- 19288175 TI - Zebrafish as a new animal model to study lymphangiogenesis. AB - The lymphatic system is essential for fluid homeostasis, fat absorption and immune responses, and also plays key roles under pathological conditions, such as tumor metastasis, lymphoedema and inflammation. The main function of the lymphatic vascular system is to return excess interstitial fluid back to the blood vascular system. Lymph, including fluid, macromolecules, leukocytes and activated antigen-presenting cells, is transported from the blind-ended lymphatic capillaries toward the collecting lymphatic vessels; for there, it is returned to the blood circulation through lymphatico-venous junctions (Alitalo et al. in Nature 438:946-954, 2005). Despite its importance, lymphangiogenesis remains poorly understood. The lack of specific markers has complicated the identification of lymph vessels, and a small animal model that could be genetically manipulated to discover the function of novel lymphangiogenic candidates has only recently become available (Ny et al. in Nat Med 11(9):998 1004, 2005). Since 2004, we have worked to make the zebrafish a new genetic model for unraveling the function of candidate genes involved in lymphangiogenesis. We have demonstrated that zebrafish possess a lymphatic vascular system that shares the morphological, molecular and functional characteristics of the lymphatic vessels found in other vertebrates (Yaniv et al. in Nat Med 12(6):711-716, 2006). In this process, we realized that it was necessary to seek a common definition for the lymph system which would be applicable from fish to man. The aim of this article is to review classical, mainly morphological, studies in order to elucidate the nature of the lymphatic system. PMID- 19288176 TI - Macroprolactinomas presenting as nasal polyps: a series of three cases. AB - BACKGROUND: Pituitary tumours that present with nasal symptoms are uncommon. Management can be difficult due to their aggressive nature, location and extension. METHODS: We report a series of three cases of prolactinomas that enlarged inferiorly presenting initially as nasal polyps. RESULTS: Recurrence of symptoms (case 1) prompted testing for serum prolactin and examination of histology confirmed the presence of a prolactinoma. In cases 2 and 3, radiological evidence of a pituitary mass prompted testing for a prolactinoma. No patients exhibited clinical signs of hyperprolactinaemia. All three cases have residual tumour at 2-4 years after diagnosis, despite prolactin levels approaching the normal range on dopaminergic therapy. CONCLUSION: Pituitary tumours that invade the nasal cavity are rare and clinicians should be aware of their existence. A prolactinoma should be considered in the differential diagnosis of nasopharyngeal tumours. Measurement of serum prolactin can expedite a diagnosis and prevent delay of treatment with dopamine agonists. PMID- 19288177 TI - Metastatic adenocarcinoma of the lung mimicking mucoid degeneration of the anterior cruciate ligament. AB - We report a case of a 63-year-old male with known adenocarcinoma of the lung who presented with knee pain which was initially diagnosed as mucoid degeneration of the anterior cruciate ligament (ACL) on magnetic resonance imaging (MRI). Due to persistent knee pain an interval MRI was performed, followed by image guided biopsy which showed metastatic adenocarcinoma of the lung infiltrating the ACL. This is the first reported case in the literature of an intra-articular metastasis mimicking mucoid degeneration of the ACL. PMID- 19288178 TI - High sensitivity C-reactive protein: a predictor for recurrence of atrial fibrillation after successful cardioversion. AB - Atrial fibrillation (AF) is often associated, more or less indirectly, with an inflammatory acute or chronic process. So it is probable that the inflammation could contribute to the genesis and the perpetuation of this dysrhythmia. Phlogistic test indexes in patients (pts) with AF will be positive and have prognostic significance in patients treated with electrical cardioversion with restoration of a sinus rhythm. We evaluated 106 pts affected by AF of recent onset without known cardiovascular disease. We measured the plasma concentration of C-reactive protein (CRP) through a high sensibility method, in addition to routine blood samples. We performed an ECG 1 week and a Holter ECG monitoring 1 and 6 months after the electrical cardioversion. The CRP values were high (5.8 +/ 10.7 U/L), with values above the normal range in 60 pts. After electrical cardioversion, we obtained restoration of sinus rhythm in all the patients. One week after cardioversion, 85 pts (80%) were in sinus rhythm, while after 6 months 60 pts (56%) maintained a sinus rhythm. In total 46 (43%) patients had a recurrence of atrial fibrillation within 6 months, and 41 of these 46 patients (89%) had elevated values of CRP (P < 0.001 with respect to the patients who maintained a sinus rhythm). 18/21 patients (86%) with an AF relapse in the first week and 23/25 patients (92%) with AF recurrences at 6 months later had elevated values of CRP. The patients with AF may have elevated values of CRP, and the assessment of this increase may be predictive of early relapses of AF after electrical cardioversion. PMID- 19288179 TI - A vivid example of angioedema. PMID- 19288180 TI - Prognostic implications of diabetes phenotyping: new concepts for an old disease. PMID- 19288181 TI - Use of D-dimer testing to determine duration of anticoagulation, risk of cardiovascular events and occult cancer after a first episode of idiopathic venous thromboembolism: the extended follow-up of the PROLONG study. AB - BACKGROUND: The PROLONG study showed that D-dimer (D-d) testing could help tailor the duration of anticoagulation after idiopathic venous thromboembolism (VTE). In this report the initial 18 month study follow-up was extended for 1 year. MATERIALS AND METHODS: D-d was measured 1 month after anticoagulation withdrawal for a first episode of idiopathic VTE. Patients with a normal D-d did not resume anticoagulation, while patients with an abnormal D-d were randomized to either resume or not resume treatment. The primary outcome was the composite of recurrent VTE and major bleeding. Secondary end-points were cardiovascular events, newly diagnosed cancers and deaths. RESULTS: D-d was abnormal in 222/608 (36.5%) patients. Average follow-up was 2.55 years. Twenty-eight events occurred in the 121 patients who stopped anticoagulation (23.1%, 9.6% person-years) and five in the 101 patients who resumed anticoagulation (5.0%, 2.0% person-years, adjusted hazard ratio-HR = 3.76; P = 0.008). Recurrence rate was higher in patients with abnormal D-d who stopped anticoagulation than in patients with normal D-d (51 events in 386 patients -13.2%; 5% person-years; adjusted HR 1.70; P = 0.045). The adjusted HR ratio associated with normal D-d versus abnormal D-d in patients who resumed anticoagulation was 2.7 (P = 0.042). An abnormal D-d was associated with a non significant higher risk of cardiovascular events and newly diagnosed cancers vs normal D-d. CONCLUSIONS: Patients with an abnormal D-d at 1 month after withdrawal of VKA have a significant risk of recurrence over a 2.55 year follow-up and they benefit from resuming anticoagulation. PMID- 19288182 TI - Involvement of multidrug resistance-associated protein 1 in intestinal toxicity of methotrexate. AB - PURPOSE: Methotrexate (MTX) causes dose-limiting gastrointestinal toxicity due to exposure of intestinal tissues, and is a substrate of the multidrug resistance associated protein (MRP) 1. Here we examine the involvement of MRP1, which is reported to be highly expressed in the proliferative crypt compartment of the small intestine, in the gastrointestinal toxicity of MTX. METHODS: MTX was intraperitoneally administered to mrp1 gene knockout (mrp1 ((-/-))) and wild-type (mrp1 ((+/+))) mice. Body weight, food and water intake were monitored, intestinal histological studies and pharmacokinetics of MTX were examined. RESULTS: mrp1 ((-/-)) mice more severely decreased body weight, food and water intake than mrp1 ((+/+)) mice. Almost complete loss of villi throughout the small intestine in mrp1 ((-/-)) mice was observed, whereas the damage was only partial in mrp1 ((+/+)) mice. Plasma concentration and biliary excretion profiles of MTX were similar in mrp1 ((-/-)) and mrp1 ((+/+)) mice, though accumulation of MTX in immature proliferative cells isolated from mrp1 ((-/-)) mice was much higher compared to mrp1 ((+/+)) mice. Immunostaining revealed localization of Mrp1 in plasma membrane of the intestinal crypt compartment in mrp1 ((+/+)) mice, but not in mrp1 ((-/-)) mice. CONCLUSION: Mrp1 determines the exposure of proliferative cells in the small intestine to MTX, followed by gastrointestinal toxicity. PMID- 19288183 TI - Longitudinal neuronal organization and coordination in a simple vertebrate: a continuous, semi-quantitative computer model of the central pattern generator for swimming in young frog tadpoles. AB - When frog tadpoles hatch their swimming requires co-ordinated contractions of trunk muscles, driven by motoneurons and controlled by a Central Pattern Generator (CPG). To study this co-ordination we used a 3.5 mm long population model of the young tadpole CPG with continuous distributions of neurons and axon lengths as estimated anatomically. We found that: (1) alternating swimming-type activity fails to self-sustain unless some excitatory interneurons have ascending axons, (2) a rostro-caudal (R-C) gradient in the distribution of excitatory premotor interneurons with short axons is required to obtain the R-C gradient in excitation and resulting progression of motoneuron firing necessary for forward swimming, (3) R-C delays in motoneuron firing decrease if excitatory motoneuron to premotor interneuron synapses are present, (4) these feedback connections and the electrical synapses between motoneurons synchronise motoneuron discharges locally, (5) the above findings are independent of the detailed membrane properties of neurons. PMID- 19288184 TI - Mechanisms and applications of theta-burst rTMS on the human motor cortex. AB - Theta-burst Stimulation (TBS) is a novel form of repetitive transcranial magnetic stimulation (rTMS). Applied over the primary motor cortex it has been successfully used to induce changes in cortical excitability. The advantage of this stimulation paradigm is that it is able to induce strong and long lasting effects using a lower stimulation intensity and a shorter time of stimulation compared to conventional rTMS protocols. Since its first description, TBS has been used in both basic and clinical research in the last years and more recently it has been expanded to other domains than the motor system. Its capacity to induce synaptic plasticity could lead to therapeutic implications for neuropsychiatric disorders. The neurobiological mechanisms of TBS are not fully understood at present; they may involve long-term potentiation (LTP)- and depression (LTD)-like processes, as well as inhibitory mechanisms modulated by GABAergic activity. This article highlights current hypotheses regarding the mechanisms of action of TBS and some central factors which may influence cortical responses to TBS. Furthermore, previous and ongoing research performed in the field of TBS on the motor cortex is summarized. PMID- 19288185 TI - Evidence that human blastomere cleavage is under unique cell cycle control. AB - PURPOSE: To understand the molecular pathways that control early human embryo development. METHODS: Improved methods of linear amplification of mRNAs and whole human genome microarray analyses were utilized to characterize gene expression in normal appearing 8-Cell human embryos, in comparison with published microarrays of human fibroblasts and pluripotent stem cells. RESULTS: Many genes involved in circadian rhythm and cell division were over-expressed in the 8-Cells. The cell cycle checkpoints, RB and WEE1, were silent on the 8-Cell arrays, whereas the recently described tumor suppressor, UHRF2, was up-regulated >10-fold, and the proto-oncogene, MYC, and the core element of circadian rhythm, CLOCK, were elevated up to >50-fold on the 8-Cell arrays. CONCLUSIONS: The canonical G1 and G2 cell cycle checkpoints are not active in totipotent human blastomeres, perhaps replaced by UHRF2, MYC, and intracellular circadian pathways, which may play important roles in early human development. PMID- 19288186 TI - Brn-3a deficiency transiently increases expression of calbindin D-28 k and calretinin in the trigeminal ganglion during embryonic development. AB - Immunohistochemistry for neuron-specific nuclear protein (NeuN), caspase-3, calcitonin gene-related peptide (CGRP), and calcium-binding proteins was performed on the trigeminal ganglion (TG) in wild type and Brn-3a knockout mice at embryonic days 12.5-16.5 (E12.5-E16.5). In Brn-3a knockout mice, the number of NeuN-immunoreactive (ir) neuron profiles increased at E14.5 (40.0% increase) and decreased at E16.5 (28.3% reduction) compared to wild type mice. Caspase-3-ir neuron profiles were abundant in the TG of wild type mice at E12.5-E16.5. However, the loss of Brn-3a decreased the number of caspase-3-ir neuron profiles at E12.5 (69.7% reduction) and E14.5 (51.7% reduction). At E16.5, the distribution of caspase-3-ir neuron profiles was barely affected by the deficiency. CGRP-ir neuron profiles were observed in the TG of wild type mice but not knockout mice at E12.5. At E14.5 and E16.5, CGRP-ir neuron profiles were abundant in both wild type and knockout mice. Calbindin D-28 k (CB)-ir neuron profiles decreased in the TG of mutant mice at E12.5 compared to wild type mice (56.4% reduction). At E14.5, however, Brn-3a deficiency transiently increased CB ir neuron profiles (169.4% increase as compared to wild type mice). Calretinin (CR)-ir neuron profiles could not be detected in the TG of wild type mice at E12.5-16.5. However, numerous CR-ir neuron profiles transiently appeared in the knockout mouse at E14.5. Parvalbumin (PV)-ir neurons appeared in wild type and knockout mice at E14.5. At this stage, the number of large (>50 mum(2)) PV-ir neuron profiles in knockout mice was fewer than that in wild type mice. The number and cell size of PV-ir neuron profiles were barely affected by the deficiency at E16.5. The present study indicates that the loss of Brn-3a causes increase of TG neurons at E14.5 and decrease of TG neurons at E16.5. It is also suggested that Brn-3a deficiency affects the number and cell size of CGRP- and calcium-binding protein-containing neurons at E12.5 and E14.5. Caspase-3 dependent cell death of CB- and CR-ir neurons may be suppressed by the deficiency at E14.5. PMID- 19288187 TI - Alterations induced by ischemic preconditioning on secretory pathways Ca2+-ATPase (SPCA) gene expression and oxidative damage after global cerebral ischemia/reperfusion in rats. AB - Ischemic preconditioning (IPC) represents the phenomenon of CNC adaptation, which results in increased tolerance of CNS to lethal ischemia. Brain ischemia/reperfusion (IRI) initiates a catastrophic cascade in which many subcellular organelles play an important role. The Golgi apparatus, which is a part of secretory pathways (SP), represents the Ca(2+) store and regulates secretion of proteins for growth/reorganization of neuronal circuit by secretory Ca(2+)ATPases (SPCA1). The purpose of this study is to evaluate the effect of IRI and preconditioning on SPCA1 gene expression and oxidative damage after 4-vessel occlusion for 15 min and after being exposed to different reperfusion periods. Rats were preconditioned by 5 min of sub-lethal ischemia and 2 days later, 15 min of lethal ischemia was induced. Our experiments conclusively showed IRI-induced depression of SPCA activity and lipo- and protein oxidation in rat hippocampal membranes. IRI also activates the induction of SPCA1 gene expression in later reperfusion periods. IPC partially suppresses lipo- and protein oxidation in hippocampal membranes and leads to partiall rovery of the ischemic-induced depression of SPCA activity. In addition, IPC initiates earlier cellular response to the injury by the significant elevation of mRNA expression to 142% comparing to 1 h of corresponding reperfusion and to 11% comparing to 24 h of corresponding reperfusion, respectively. Similar patterns were observed on the translational level by Western blot analysis. Our results indicate the specific SPCA1 expression pattern in ischemic hippocampus. It also shows that the SPCA expression and the post-translational changes induced by ischemia are modulated by the IPC. This might serve to understand the molecular mechanisms involved in the structural integrity and function of the SP after ischemic challenge. It also suggests that there is a correlation of SPCA function with the role of SP in the response to pre-ischemic challenge. PMID- 19288188 TI - Flow cytometry analysis of neural differentiation markers expression in human glioblastomas may predict their response to chemotherapy. AB - Glioblastoma multiforme (GBM) represents an extremely chemoresistant tumour type. Here, authors analysed the immunophenotype of GBM tumours by flow cytometry and correlated the immunophenotypic characteristics with sensitivity to chemotherapy. The expression of selected neural and non-neural differentiation markers including A2B5, CD34, CD45, CD56, CD117, CD133, EGFR, GFAP, Her-2/neu, LIFR, nestin, NGFR, Pgp and vimentin was analysed by flow cytometry in eleven GBM (WHO gr.IV) patients. The sensitivity of tumour cells to a panel of chemotherapeutic agents was tested by the MTT assay. All tumours were positive for A2B5, CD56, nestin and vimentin. CD133, EGFR, LIFR, NGFR and Pgp were expressed only by minor tumour cell subpopulations. CD34, CD45, CD117, GFAP and Her-2/neu were constantly negative. Direct correlations were found between the immunophenotypic markers and chemosensitivity: A2B5 vs lomustine (r(2) = 0.642, P = 0.033), CD56 vs cisplatin (r(2) = 0.745, P = 0.013), %Pgp(+) vs vincristine (r(2) = 0.846, P = 0.008), and %NGFR(+) vs daunorubicine (r(2) = 0.672, P = 0.047) and topotecan (r(2) = 0.792, P = 0.011). In contrast, inverse correlations were observed between: EGFR vs paclitaxel (r(2) = -0.676, P = 0.046), CD133 vs dacarbazine (r(2) = -0.636, P = 0.048) and LIFR vs daunorubicine (r(2) = -0.878, P = 0.004). Finally, significant associations were also found among sensitivities to different chemotherapeutic agents and among different immunophenotypic markers. In conclusion, histopathologically identical GBM tumours displayed a marked immunophenotypic heterogeneity. The expression of A2B5, CD56, NGFR and Pgp appeared to be associated with chemoresistance whereas CD133, EGFR and LIFR expression was characteristic of chemosensitive tumours. We suggest that flow cytometric imunophenotypic analysis of GBM may predict chemoresponsiveness and help to identify patients who could potentially benefit from chemotherapy. PMID- 19288189 TI - Glypican-3 reexpression regulates apoptosis in murine adenocarcinoma mammary cells modulating PI3K/Akt and p38MAPK signaling pathways. AB - Glypican-3 (GPC3) is a proteoglycan involved in proliferation and cell survival. Several reports demonstrated that GPC3 is downregulated in some tumors, such as breast cancer. Previously, we determined that GPC3 reexpression in the murine mammary adenocarcinoma LM3 cells induced an impairment of their invasive and metastatic capacities, associated with a decrease of their motility and an increase of their cell death. We demonstrated that GPC3 inhibits canonical Wnt signaling, as well as it activates non canonical pathway. Now, we identified signaling pathways responsible for the pro-apoptotic role of GPC3 in LM3 cells. We found for the first time that GPC3 inhibits the PI3K/Akt anti-apoptotic pathway while it stimulates the p38MAPK stress-activated one. We report a concomitant modulation of CDK inhibitors as well as of pro- and anti-apoptotic molecules. Our results provide new clues regarding the mechanism involved in the modulation induced by GPC3 of mammary tumor cell growth and survival. PMID- 19288190 TI - A novel BRCA2 mutation that segregates with breast and prostate cancer in a Spanish family. AB - We report a novel germline 5369delATTT mutation in BRCA2 gene, detected in a 45 year-old woman with bilateral breast cancer. This deletion was also detected in her father with prostatic cancer and her sister with breast cancer. The mutation originates a premature stop at codon 1723 of BRCA2 protein and has not been documented in any published report to the best of our knowledge. PMID- 19288191 TI - Up-regulation of epithelial membrane protein-1 in the temporal neocortex of patients with intractable epilepsy. AB - Epithelial membrane protein-1 (EMP-1), called Tumor-associated membrane protein, is the marker of a drug-resistant tumor and take part in the drug-resistant mechanism of tumor, with the relationship of epidermal growth factor receptor (EGFR). Because there are some similarities between the pathogenesis and the drug resistance mechanism of tumor and the drug resistance mechanisms in epilepsy. EMP1 expression may be connected with the drug-resistance mechanism of epilepsy. We detected EMP-1 by gene scanning and immunohistochemistry staining, comparing the IE group and the control group, and we investigated the relationship between EMP-1 and EGFR by double-label immunofluorescence staining in the IE group. We found expression of EMP-1 mRNA was higher in IE per the gene scanning, EMP-1 immunoreactivity was apparent in neurons of IE patients but not in the control group, and the expression of EMP-1 and EGFR occurred in the same neuron. We confirm EMP-1 is abnormally expressed in IE and suggest the interaction of EGFR and EMP-1 plays a role in the mechanism of drug resistance in epilepsy and may be a new gene for drug resistance. PMID- 19288192 TI - Possible involvement of extracellular ATP-P2Y purinoceptor signaling in ischemia induced tolerance of astrocytes in culture. AB - Extracellular adenosine 5'-triphosphate (ATP) activates specific G protein coupled purinoceptors (P2Y), and ATP-P2Y signaling pathways induces intracellular Ca(2+) mobilization resulting in changes in the gene expression of a variety of proteins in astrocytes. This study investigated whether the exposure of cultured astrocytes to sublethal ischemia produced resistance to subsequent lethal ischemic stress, and if so, whether the extracellular ATP-P2Y signaling pathways were responsible for the tolerance. Ischemia-like insults, sublethal oxygen glucose deprivation (sOGD), produced tolerance to subsequent lethal OGD stress in cultured astrocytes. Early during reperfusion after sOGD, the amount of extracellular ATP and the expression of both P2Y(1) and P2Y(2) receptors were increased, leading to enhanced activation of the extracellular ATP-P2Y signaling pathways. The occurrence of intracellular spontaneous Ca(2+) oscillations was also increased. In addition, sOGD treatment enhanced the expression of the phosphorylated form of extracellular signal-regulated protein kinases 1 and 2 (p ERK 1/2), and treatment with an inhibitor of ERK significantly attenuated the sOGD-induced ischemic tolerance of astrocytes. PMID- 19288193 TI - Enhanced 5-HT(2A) receptors in brain stem and ALDH activity in brain stem and liver: 5-HT(2A) regulation on ALDH in primary hepatocytes cultures in vitro. AB - Brain serotonin (5-HT) modulates the neural effects of ethanol. In the present study, we investigated the changes in 5-HT level, 5-HT(2A) receptor binding and aldehyde dehydrogenase (ALDH) activity in brain stem and liver of ethanol treated rats and 5-HT(2A) regulation on ALDH in hepatocyte cultures in vitro. The 5-HT content in the brain stem and liver significantly decreased with an increased 5 HIAA/5-HT ratio in the ethanol treated rats compared to control. Scatchard analysis of [(3)H] (+/-)2,3-dimethoxyphenyl-1-[2-(-4-piperidine)-methanol] [(3)H] MDL 100907 against ketanserin in brain stem of ethanol treated rats showed a significant increase in B (max) without any change in K (d) compared to control. The competition curve for [(3)H] MDL 100907 against ketanserin fitted one-site model in both control and ethanol treated rats with unity as Hill slope value. A significant increase in V (max) of ALDH activity in liver and a significant decrease in K (m) in liver and brain stem of ethanol treated rats compared to control was observed. In 24 h culture studies, an increase in enzyme activity was observed in cells in medium with 10% ethanol. The elevated ALDH activity in ethanol treated cells was reversed to control level in presence of 10(-5) and 10( 7) M 5-HT. Ketanserin, an antagonist of 5-HT(2A), reversed the effect of 5HT on 10% ethanol induced ALDH activity in hepatocytes. Our results showed that there was a decreased 5-HT content with an enhanced 5-HT(2A) receptor and aldehyde dehydrogenase activity in the brain stem of alcohol treated rats and in vitro hepatocyte cultures. The enhanced ALDH activity in ethanol supplemented hepatocytes was reversed to control level in presence of 10(-5) and 10(-7) M 5 HT. PMID- 19288194 TI - Use of recombinant factor VIIa to correct the coagulation status of individuals with advanced liver disease prior to a percutaneous liver biopsy. AB - OBJECTIVES: Percutaneous liver biopsies are used to grade and stage liver disease and are also useful in monitoring the progress of liver disease over time as well as the response to medical therapies. The present study was undertaken to assess the effectiveness of recombinant factor VIIa as a means of transiently correcting the coagulopathy, enabling the safe performance of a percutaneous liver biopsy in patients in whom the use of fresh-frozen plasma is not possible without precipitating pulmonary edema or who have a treatment induced (iatrogenic) coagulopathy. METHODS: The subjects of this report consisted of 18 consecutive individuals with advanced disease induced, and 15 with a therapeutic iatrogenic induced, coagulopathy. All biopsies were performed by a single hepatologist. Before and 6 h after each biopsy, a prothrombin time and partial thromboplastin time was obtained from each subject. Mean values +/- the standard error of the mean were obtained using the independent samples T-test. RESULTS: Recombinant factor VIIa had a marked effect in transiently correcting the mean prothrombin time in these subjects allowing for a safe complication free percutaneous biopsy in this high-risk group. CONCLUSIONS: Recombinant factor VIIa could be used to obtain a clinically indicated liver biopsy in severely ill patients, who without this therapeutic agent, would either not be biopsied or, if biopsied, would require much longer hospitalization and the use of fresh-frozen plasma (with its risks of volume overload and infection). PMID- 19288195 TI - "Stat" medication administration predicts hospital discharge. AB - OBJECTIVE: This study describes the use of "stat" medications for inpatients in a large state psychiatric hospital system, and examines the relationship between receipt of a "stat" for agitation and subsequent hospital discharge. METHODS: Use of "stat" medications in 2005 was retrospectively determined using a database that contains diagnosis and prescription information from 17 state-run adult civil facilities. A logistic regression model explored the relationship between receipt of a "stat" order for intramuscular preparations of either antipsychotics or lorazepam within the first 30 days of hospitalization and likelihood of hospital discharge by 6 months. RESULTS: Among 7,202 patients who received antipsychotic medication in 2005, 3,240 (45%) also received a "stat" psychotropic medication during that year. Among 40,651 stat orders, 19,142 (47%) were for intramuscular antipsychotics or lorazepam presumably given for the treatment of agitation. Among 1,673 patients admitted in the first 6 months of 2005, 415 (25%) received at least one such "agitation stat." The percent discharged at six months among "agitation stat" receivers was 39%, compared to 69% among those who did not receive an "agitation stat" (chi-square = 115, df = 1, P < .001). Regression analysis showed that receiving an "agitation stat" in the first 30 days of hospitalization was associated with a 37% lower likelihood of being discharged by 6 months after admission (odds ratio .63, 95% CI: .46-.86). CONCLUSIONS: "Stat" medications are commonly used. The use of "agitation stat" medications can be used as a proxy for clinical stability and may prove to be a useful outcome measure for future pharmacoepidemiologic studies of comparative medication effectiveness. PMID- 19288196 TI - Evaluating mediation in longitudinal multivariate data: mediation effects for the Aban Aya Youth Project drug prevention program. AB - This study illustrates a method to evaluate mediational mechanisms in a longitudinal prevention trial, the Aban Aya Youth Project (AAYP). In previous studies, interventions of AAYP were found to be effective in reducing the growth of violence, substance use and unsafe sex among African American adolescents. In this article, we hypothesized that the effects of the interventions in reducing the growth of substance use behavior were achieved through their effects in changing intermediate processes such as behavioral intentions, attitudes toward the behavior, estimates of peers' behaviors, best friends' behaviors, and peer group pressure. In evaluating these mediational mechanisms, difficulties arise because the growth trajectories of the substance use outcome variable and some of the mediating variables were curvilinear. In addition, all of the multivariate mediational measures had planned missing data so that a score from the multiple items for a mediator could not be formed easily. In this article, we introduce a latent growth modeling (LGM) approach; namely, a two-domain LGM mediation model, in which the growth curves of the outcome and the mediator are simultaneously modeled and the mediation effects are evaluated. Results showed that the AAYP intervention effects on adolescent drug use were mediated by normative beliefs of prevalence estimates, friends' drug use behavior, perceived friends' encouragement to use, and attitudes toward the behavior. PMID- 19288197 TI - Novel approach of predicting fracture load in the human proximal femur using non invasive QCT imaging technique. AB - This paper presents an analysis of predicting the load-bearing capacities of human femurs using quantitative computer tomography (QCT)-based beam theory. Cross-sectional images of 12 human cadaver femurs (intact bones, age: 39-77 years; male = 8, female = 4) were scanned in conjunction with a calcium hydroxyapatite phantom which has five chambers of known densities. The apparent densities obtained from the scans were used to evaluate the Young's modulus (E) by applying the established empirical relationships. The fracture load of a configuration that simulated single-legged stance was measured experimentally and compared with the predicted failure load using a composite beam theory, plane stress model of the femur. In this model, the failure was assumed to occur at the weakest cross-section through the bone determined from QCT-based structural analysis. In contrast to the other experimental investigations, the setup used in this study considers the entire length of a human femur and also incorporates a novel mechanical jig to mimic the realistic physiological scenario. In one of our earlier studies, simulated lytic defects of varying size were created at the inter-trochanteric region of femurs and their load-bearing capacities were calculated based on their structural properties. Both the results obtained from the current study as well as the ones from our previous study were used to assess the viability of the methodology. A high degree of correlation was observed when the predicted failure loads obtained from the intact femurs and previously studied defective femurs were compared with the ex vivo fracture loads. The coefficients of determination (R(2)) of QCT-derived predicted loads with respect to the measured failure loads were 0.80 for the intact femurs and 0.87 for the defective femurs. The results suggest that the QCT-derived beam analysis provides a viable approach for the assessment of load-bearing capacity in various clinical scenarios. PMID- 19288198 TI - Episcopal measure of faith tradition: a context-specific approach to measuring religiousness. AB - Precise measurement of religiousness remains a vexing problem. In addition to relying almost exclusively on self-report, existing measures of religiousness pay little attention to the specific context of religious belief, and this may override distinctive norms of particular faith traditions and potentially confound the conclusions drawn from such research. To address these limitations, the authors describe a modified form of narrative content analysis that could eventually sort respondents into distinct theological traditions. A pilot test among Episcopalians demonstrates encouraging reliability (kappa 0.74, 95% LCI 0.47, P < 0.0002), and tests for convergent and discriminate validity suggest that the context of religious belief is both relevant and insufficiently assessed by the existing paradigm of religious measurements. If validated in a religiously diverse sample, this approach could be combined with existing, context-free measures of religiousness to generate more meaningful findings. PMID- 19288199 TI - A novel 3-D model for cell culture and tissue engineering. AB - A novel method of making microcapsules in a macrocapsule is demonstrated as a 3-D culture system in this article. Mouse embryonic stem (mES) cells as model cells were used in the 3-D culture space, and the cell viability and histological observation were conducted. Furthermore, Oct4 gene expression was evaluated for the undifferentiated status of mES cells in this 3-D model. The results showed that mES cells can grow in this 3-D model and retain their normal viability and morphology. This 3-D model allows mES cells to stay in the undifferentiated state better than 2-D culture systems. This work demonstrates a new 3-D tissue model which can provide an in vivo like microenvironment for non-differentiated mES cells with good immunoisolation. This approach may bridge the gap between traditional 2-D cell culture and animal models. PMID- 19288200 TI - Successful priority setting in low and middle income countries: a framework for evaluation. AB - Priority setting remains a big challenge for health managers and planners, yet there is paucity of literature on evaluating priority setting. The purpose of this paper is to present a framework for evaluating priority setting in low and middle income countries. We conducted a qualitative study involving a review of literature and Delphi interviews with respondents knowledgeable of priority setting in low and middle income countries. Respondents were asked to identify the measures of successful priority setting in low and middle income countries. Responses were grouped as: immediate internal or external/delayed internal or external. We also identified some pre-requisites for successful priority setting. The immediate internal measures included increased efficiency in decision making, improved quality of decisions and fairer priority setting. Immediate External measures included-improved public understanding and acceptance of decisions, increased public participation, increased trust. Delayed Internal measures included increased satisfaction, understanding, compliance, balanced budget, achievement of organization goals, and improved internal accountability. Delayed External measures include impact on policy and practice, improved population health and reduction of health inequalities, achievement of health system goals and strengthening of health care systems. Identified pre-requisites for successful priority setting included; the presence of credible priority setting institutions, incentives for participation and implementation and resources, capacity and political will to implement. These would be augmented in a conducive political, social and economic context. This framework, although not exhaustive, provides a practical basis for planning and evaluating priority setting in low and middle income countries. PMID- 19288201 TI - Public perceptions of ethical issues regarding adult predictive genetic testing. AB - The purpose of this study was to explore the views of members of the general public regarding ethical issues in adult predictive genetic testing. The literature pertaining to ethical issues regarding to adult predictive genetic testing is largely restricted to the views of 'experts' who have emphasized informed consent, patent issues, and insurance discrimination. Occasionally the views of patients who have undergone genetic counselling and testing have been elicited, adding psychosocial and family issues. However, the general public has not had the opportunity to contribute. In order to explore theatre as a health policy research tool, 1,200 audience members attended the play 'Sarah's Daughters' in seven Canadian cities, following which audience discussions were audiotaped. This study performed a secondary qualitative analysis of the data to identify the ethical issues of adult predictive genetic testing important to members of the general public. The identified issues were: (1) need for public education; (2) choice to undergo genetic counselling and testing; (3) access to genetic counselling and testing; and (4) obligations regarding the handling of genetic information. Audience members emphasized public education and access to information regarding potential choices, which was different from the emphasis on informed consent and other ethical issues prominent in the literature. Members of the general public emphasized ethical issues that were different than those identified by experts and patients. It is essential that members of the public be included in complex and controversial public policy decisions. PMID- 19288202 TI - HIV regulation of amyloid beta production. AB - The use of antiretroviral therapy for HIV infection has extended the survival of individuals living with HIV. However, the effects of chronic HIV infection and aging are introducing another facet of HIV complications. HIV therapy can calm the immune system and lower viral replication to undetectable but the virus is still present. In the brain, amyloid beta (Abeta) increases during normal aging but Abeta accumulation appears to accelerate in HIV infection. HIV Tat protein inhibits the major Abeta-degrading enzyme neprilysin with the cysteine-rich domain of Tat being essential for this inhibition. In this minireview, we also include new data that the beta chemokine, CCL2/MCP-1, associated with HIV migration to the brain, also causes an increase in Abeta. These findings may explain the continued cognitive dysfunction found in HIV-infected individuals controlled on antiviral therapy. PMID- 19288203 TI - Effective modification of particle surface properties using ultrasonic water mist. AB - The goal of the present study was to design a new technique to modify particle surface properties and, through that, to improve flowability of poorly flowing drug thiamine hydrochloride and pharmaceutical sugar lactose monohydrate of two different grades. The powdered particles were supplied by a vibratory feeder and exposed to an instantaneous effect of water mist generated from an ultrasound nebulizer. The processed and original powders were evaluated with respect to morphology (scanning electron microscopy, atomic force microscopy, and spatial filtering technique), flow, and solid state properties. It was found that rapid exposition of pharmaceutical materials by water mist resulted in the improvement of powder technical properties. The evident changes in flowability of coarser lactose were obviously due to smoothing of particle surface and decreasing in the level of fines with very slight increment in particle size. The changes in thiamine powder flow were mainly due to narrowing in particle size distribution where the tendency for better flow of finer lactose was related to surface and size modifications. The aqueous mist application did not cause any alteration of the crystal structures of the studied materials. The proposed water mist treatment technique appears to be a robust, rapid, and promising tool for the improvement of the technological properties of pharmaceutical powders. PMID- 19288204 TI - Taking a risk perception approach to improving beach swimming safety. AB - BACKGROUND: Beach swimming is generally associated with a healthy lifestyle, yet the risk of drowning is universally significant. PURPOSE: The purpose of the study was to investigate the factors associated with safe swimming behavior using protection motivation theory as a guiding theoretical framework. METHOD: This cross-sectional study surveyed a sample of beachgoers (N = 3371) aged > or =16 years who completed an anonymous, self-report questionnaire to assess the associations between perceptions of the risk of drowning and safe swimming behavior. RESULTS: Compared with males, females perceived greater severity, vulnerability, response efficacy, and concern regarding their risk of drowning. Males, Maori, and 16 to 29 year olds reported higher self-efficacy scores compared to females, other ethnic groups, and older participants, respectively. After controlling for confounding variables, people perceiving a greater threat (severity) of experiencing difficulty while swimming as well as those reporting higher response efficacy (beliefs about the effectiveness of drowning prevention measures) were more likely to report safe swimming behavior. CONCLUSION: The effectiveness of water safety education programs could be strengthened by enhancing risk appraisal and coping skills and counter-acting the tendency of males and younger adults to overestimate their swimming ability and underestimate their risk with regard to drowning. PMID- 19288205 TI - Dimensions of social support and depression in patients at increased psychosocial risk recovering from myocardial infarction. AB - BACKGROUND: There is considerable evidence that depression and low social support are associated with increased morbidity and mortality for patients with coronary heart disease (CHD). However, there is a lack of consensus regarding the measurement of social support and its relation to depression. PURPOSE: The primary purpose of the present study was to identify key dimensions of existing social support and depression measures for patients with CHD using factor analysis. METHOD: Seven hundred-five patients with a recent acute myocardial infarction and either depression, low social support, or both, completed measures of several types of social support and depression. Exploratory and confirmatory factor analysis were used to examine the underlying dimensions of the existing social support and depression measures, and to compare theoretically plausible models specifying the relation between the social support and depression factors. RESULTS: Confirmatory factor analysis indicated that an approach in which smaller facets of depression are measured (somatic, cognitive/affective, anxious) and social support (perceived emotional support from intimate relationships; perceived tangible support from peripheral contacts; and the number of children, relatives, and friends in a patient's support network), may be the most optimal way to measure social support and depression in this population RMSEA = 0.05; CFI = 0.81; TLI = 0.88). CONCLUSION: Efforts to identify patients at increased psychosocial risk may be improved by screening for these subcomponents of social support and depression. PMID- 19288206 TI - Salivary cortisol and binge eating disorder in obese women after surgery for morbid obesity. AB - BACKGROUND: Binge eating episodes characterized by loss of control are hypothesized to be accompanied by changes in hypothalamic pituitary adrenal (HPA) axis functioning. Cortisol is an end product of this neuroendocrine stress system. PURPOSE: The aim of this study was to examine the cortisol levels and the awakening cortisol response (ACR) in obese persons showing binge eating after surgery for morbid obesity. METHOD: Sixteen obese women with binge eating disorder (BED) and 18 obese women without BED participated in the study. Means+/ SD: age 43 +/- 15, body mass index 40 +/- 8. Salivary cortisol, anthropometric assessments, and the eating disorder examination interview were taken. RESULTS: Women with BED showed a significantly lower waist-to-hip ratio and cortisol levels during the day than women without BED, whereas the ACR did not differ. CONCLUSION: Our cross-sectional study in a small sample generates the hypothesis that neuroendocrine regulation differs between obese women with and without BED after obesity surgery. This finding needs replication in future studies that should also examine the causal direction of the observed association. PMID- 19288207 TI - Identifying factors associated with good health and ill health : not just opposite sides of the same coin. AB - BACKGROUND: Work-related health research has traditionally focused on identifying risks rather than determinants of good health. Our knowledge of variation in ill health is thus greater than our understanding of such variations in good health. PURPOSE: In this study, the associations between work-environment exposures and good health are examined. We are especially interested in contrasting our indices of ill health with a narrow measure of good health. Moreover, the salutary effect of sense of coherence (SOC) is explored, focusing particularly on its moderating role. METHOD: Data stem from the panel of Swedish level of living surveys for 1991 and 2000. The analysis is based on a sample of 2,334 employed men and women. Logistic regressions are used. RESULTS: Assessed work-environment factors are to a large extent related, in a mirrored way, to good health and ill health. The models' fit are, however, generally better for the latter. Our findings also indicate that SOC has a protective role for individuals exposed to work risks such as stress and high physical demands. CONCLUSION: To improve our understanding of what promotes good health, research needs to focus on salutary factors. One such salutary factor explored in this paper is sense of coherence. PMID- 19288208 TI - Cross-cultural comparison of lack of regular physical activity among college students: Universal versus transversal. AB - PURPOSE: This study examined cultural influence on personal and behavioral correlates of lack of regular physical activity (PA) among college students in four countries, i.e., the United States, Costa Rica, India, and South Korea. METHOD: Public universities were randomly chosen among the four countries. A total of 4,685 students participated in the study during the 2006-2007 academic year with a response rate of 90.1%. The vast majority of the questions on the instrument were adopted from the Youth Risk Behavior Survey and the Behavioral Risk Factor Surveillance System questionnaires. The instrument was translated into Spanish and Korean and then back-translated into English to check accuracy of the translation. RESULTS: Low fruit consumption was a culture-universal predictor of lack of regular PA. Gender, perceived body weight, vegetable consumption, and cigarette smoking were culture-specific predictors, indicating PA might be a transversal value. Body mass index, binge drinking, and TV/video watching were not associated with lack of regular PA in any of the four countries. CONCLUSION: While PA is valued across different segments of many cultures, given the several culture-specific predictors, PA appears to be more transversal than universal. Therefore, culturally sensitive interventions are necessary to promote PA among young adults. PMID- 19288209 TI - The psychosocial work environment and maternal postpartum depression. AB - BACKGROUND: Postpartum depression is a debilitating mental disorder affecting women after childbirth. This study examined the correlates of postpartum depression at 11 weeks after childbirth, focusing on work-related stressors and applying the job demand-control-support model. METHOD: Investigators recruited a prospective cohort of 817 employed Minnesota women when hospitalized for childbirth in 2001. Trained interviewers collected data in person and by telephone at enrollment and 5 and 11 weeks postpartum from three Minneapolis and St. Paul hospitals. RESULTS: Results of hierarchical regression analysis showed that worse depression scores (Edinburgh Postnatal Depression Scale) were associated with higher psychological demands, lower schedule autonomy, and lower perceived control over work and family. Perceptions of control mediated the relationships of coworker support and schedule autonomy with postpartum depression scores. Study findings showed no significant buffering effects for decision latitude; however, coworker support and decision latitude appear to act as functional substitutes in reducing postpartum depressive symptoms. CONCLUSION: These findings raise questions about the applicability of the job demand-control support model to postpartum women or to postpartum depression. Future research could assess the impact of the interaction between the work and home environment on maternal postpartum depression. PMID- 19288210 TI - Socioeconomic status and the course of quality of life in older patients with coronary heart disease. AB - BACKGROUND: Previous research suggests that socioeconomic status (SES) might be related to the course of quality of life (QoL) in coronary heart disease (CHD) patients. The authors sought to determine whether there are differences in the course of QoL before and after the incidence of CHD among older persons of differing SES. METHOD: Two hundred two CHD patients were followed up longitudinally using a community-based survey. Data on patients' QoL were collected before the diagnosis and at three follow-up assessments. RESULTS: High SES patients reported better outcomes at the premorbid assessment with fewer depressive feelings and better physical functioning. In physical functioning, similar results were repeated 6 and 12 months after the diagnosis. Additionally, high SES patients showed better role and social functioning 1 year after CHD. A multivariate analysis of variance revealed differential longitudinal pathways in relation to SES in role, social, and physical functioning. CONCLUSION: CHD modulates premorbid differences in depressive feelings. Conversely, high SES leads to better outcomes in all functional domains in the long-term after diagnosis. Postmorbid differences in physical functioning are not directly related to CHD, but rather the reestablishment of a premorbid situation. In contrast, socioeconomic inequalities in social and role functioning are a direct response to the impact of the disease. PMID- 19288211 TI - Determination of the pKa value of the hydroxyl group in the alpha hydroxycarboxylates citrate, malate and lactate by 13C NMR: implications for metal coordination in biological systems. AB - Citric acid is an important metal chelator of biological relevance. Citric acid helps solubilizing metals, increasing their bioavailability for plants and microbes and it is also thought to be a constituent of both the extracellular and cytoplasmic low molecular iron pools occurring in plants and vertebrates. Metal coordination by citric acid involves coordination both by the carboxylate and hydroxyl groups, of particular interest is its alpha-hydroxycarboxylate function. This structural feature is highly conserved in siderophores produced by evolutionarily distant species and seems to confer specificity toward Fe(III) binding. In order to understand the mechanism of metal coordination by alpha hydroxycarboxylates and correctly evaluate the respective complex stability constants, it is essential to improve the knowledge about the ionisation of the alcohol group in these compounds. We have evaluated the hydroxyl pKa value of citric, malic and lactic acids with the objective of understanding the influence of alpha-carbon substitution. Studies at high pH values, utilizing (13)C NMR, permitted estimation of the pKa values for the three acids. The pKa (alcohol) values (14.4 for citric acid, 14.5 for malic acid, and 15.1 for lactic acid) are considerably higher than the previously reported value for citric acid (11.6) but still lower than the value of 15.5 for methanol. A comparative analysis of the three compounds indicates that different substitutions on the alpha-carbon introduce changes to the inductive effect experienced by the hydroxyl group thereby modulating its ionisation behaviour. Comparison with the siderophore rhizoferrin, which pKa (alcohol) values were confirmed to be 10 and 11.3, suggests that intra-molecular hydrogen bonding may also aid in the hydroxyl ionisation by stabilizing the resulting anion. Studies of metal coordination by alpha-hydroxycarboxylates should take these factors into account. PMID- 19288212 TI - Overexpression of Arabidopsis damaged DNA binding protein 1A (DDB1A) enhances UV tolerance. AB - Damaged DNA Binding protein 1 (DDB1) is a conserved protein and a component of multiple cellular complexes. Arabidopsis has two homologues of DDB1: DDB1A and DDB1B. In this study we examine the role of DDB1A in Arabidopsis UV tolerance and DNA repair using a DDB1A null mutant (ddb1a) and overexpression lines. DDB1A overexpression lines showed higher levels of UV-resistance than wild-type in a range of assays as well as faster DNA repair. However a significant difference between wild-type plants and ddb1a mutants was only observed immediately following UV treatment in root length and photoproduct repair assays. DDB1A and DDB1B mRNA levels increased 3 h after UV exposure and DDB1A is required for UV regulation of DDB1B and DDB2 mRNA levels. In conclusion, while DDB1A is sufficient to increase Arabidopsis UV tolerance, it is only necessary for immediate response to UV damage. PMID- 19288213 TI - Early evolution of the MFT-like gene family in plants. AB - Angiosperm genes sharing a conserved phosphatidylethanolamine-binding (PEPB) domain have been shown to be involved in the control of shoot meristem identity and flowering time. The family is divided into three subfamilies, FT-like, TFL1 like and MFT-like. This study is focused on the evolution of the MFT-like clade, suggested to be ancestral to the two other clades. We report that the bryophyte Physcomitrella patens and the lycopod Selaginella moellendorfii contain four and two MFT-like genes respectively. Neither species have any FT or TFL1-like genes. Furthermore, we have identified a new subclade of MFT-like genes in Angiosperms. Quantitative expression analysis of MFT-like genes in Physcomitrella patens reveals that the expression patterns are circadian and reaches maximum in gametangia and sporophytes. Our data suggest that the occurrence FT and TFL1-like genes, is associated with the evolution of seed plants. Expression data for Physcomitrella MFT-like genes implicates an involvement in the development of reproductive tissues in the moss. PMID- 19288214 TI - How far can we explain the social class differential in respiratory function? A cross-sectional population study of 21,991 men and women from EPIC-Norfolk. AB - The objective of this study is to investigate the association between occupational social class and respiratory function, as measured by forced expiratory volume in one-second (FEV(1)). We examined the cross sectional relationship between lung function and social class in a population study of 21,991 men and women aged 39-79 years living in the general community in Norfolk, United Kingdom, recruited using general practice age-sex registers in 1993-1997. There was a significant socioeconomic gradient in age adjusted lung function with a difference of 0.37 in mean FEV(1) in men and 0.20 in women, respectively between social class I and V. The age adjusted OR for having poor lung function was 4.13 (95% CI 2.66-6.42) in men and 2.64 (95% CI 1.74-3.99) in women for social class V compared to I. This difference was substantially attenuated after adjustment for height, weight, smoking status, respiratory illness, educational level, living in a deprived area, physical activity and plasma vitamin C levels. There was a strong socioeconomic gradient in respiratory function. In men the gradient appeared to be largely explained by smoking status and height; in women a large part of the gradient was explained by potentially modifiable factors. This suggests that socioeconomic inequalities in respiratory function may be preventable or modifiable and highlights factors for further exploration. PMID- 19288215 TI - Examining secular trends and seasonality in count data using dynamic generalized linear modelling: a new methodological approach illustrated with hospital discharge data on myocardial infarction. AB - Time series of incidence counts often show secular trends and seasonal patterns. We present a model for incidence counts capable of handling a possible gradual change in growth rates and seasonal patterns, serial correlation, and overdispersion. The model resembles an ordinary time series regression model for Poisson counts. It differs in allowing the regression coefficients to vary gradually over time in a random fashion. During the 1983-1999 period, 17,989 incidents of acute myocardial infarction were recorded in the Hospital Discharge Registry for the county of North Jutland, Denmark. Records were updated daily. A dynamic model with a seasonal pattern and an approximately linear trend was fitted to the data, and diagnostic plots indicated a good model fit. The analysis conducted with the dynamic model revealed peaks coinciding with above-average influenza A activity. On average the dynamic model estimated a higher peak-to trough ratio than traditional models, and showed gradual changes in seasonal patterns. Analyses conducted with this model provide insights not available from more traditional approaches. PMID- 19288216 TI - Modulation of antioxidant defense by Alpinia galanga and Curcuma aromatica extracts correlates with their inhibition of UVA-induced melanogenesis. AB - Ultraviolet A (UVA) irradiation is suggested to contribute to melanogenesis through promoting cellular oxidative stress and impairing antioxidant defenses. An overproduction of melanin can be associated with melanoma skin cancer and hyperpigmentation. Therefore, developing effective antimelanogenic agents is of importance. Alpinia galanga (AG) and Curcuma aromatica (CA) are traditional medicinal plants widely used for skin problems. Hence, this study investigated the antimelanogenic effects of AG and CA extracts (3.8-30 microg/ml) by assessing tyrosinase activity, tyrosinase mRNA levels, and melanin content in human melanoma cells (G361) exposed to UVA. The roles in protecting against melanogenesis were examined by evaluating their inhibitory effects on UVA-induced cellular oxidative stress and modulation of antioxidant defenses including antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GPx), and intracellular glutathione (GSH). In addition, possible active compounds accountable for biological activities of the extracts were identified by thin layer chromatography (TLC)-densitometric analysis. Our study demonstrated that UVA (8 J/cm(2)) induced both tyrosinase activity and mRNA levels and UVA (16 J/cm(2))-mediated melanin production were suppressed by the AG or CA extracts at noncytotoxic concentrations. Both extracts were able to protect against UVA induced cellular oxidant formation and depletion of CAT and GPx activities and GSH content in a dose-dependent manner. Moreover, TLC-densitometric analysis detected the presence of eugenol and curcuminoids in AG and CA, respectively. This is the first report representing promising findings on AG and CA extract derived antityrosinase properties correlated with their antioxidant potential. Inhibiting cellular oxidative stress and improving antioxidant defenses might be the mechanisms by which the extracts yield the protective effects on UVA dependent melanogenesis. PMID- 19288217 TI - Race in ovarian cancer treatment and survival: a systematic review with meta analysis. AB - OBJECTIVE: There has long been recognition of racial disparities in cancer treatment and survival. In order to investigate the etiology of racial disparities in ovarian cancer, we undertook a systematic review of the published literature. METHODS: Focusing on North America, our search of MEDLINE, PsychInfo, and EMBASE databases recovered 513 abstracts of which 98 underwent full text screening resulting in 24 studies included in the final review. After assessing heterogeneity, results were pooled where possible in a meta-analysis using a random effects model. RESULTS: Eight articles reported treatment outcomes, nine survival, and seven both. Overall African Americans were less likely to receive any form of surgical treatment for ovarian cancer [pooled relative risk (RR) 1.17 (95% confidence interval (CI): 1.10, 1.23)] compared with white women. Although the majority of the included articles reporting survival outcomes did not control for known covariates such as medical co-morbidities or treatment, we were able to pool the unadjusted results from eight articles. Taken together the meta-analysis of 106,704 women did not find a difference in five-year survival between whites and African Americans, RR 1.07 (95% CI: 0.97, 1.18). When the results were stratified by year of cancer diagnosis, studies which captured patients prior to 1985 yielded a five-year RR of survival for whites compared to African Americans of 0.93 (95% CI: 0.89, 0.97) compared with 1.17 (95% CI: 1.05, 1.31) after 1985. CONCLUSION: These results suggest that racial disparities in ovarian cancer are not due to underlying biological differences rather to the unequal application of existing treatments. PMID- 19288218 TI - Regulatory expression of genes related to metastasis by TGF-beta and activin A in B16 murine melanoma cells. AB - TGF-beta induces epithelial-mesenchymal transition, which occurs during tumor cell invasiveness in pathological state, in limited cells. As a first step to understand the role of TGF-beta and the structurally related activin during melanoma metastasis, expression of metastasis-related genes was examined in murine melanoma cells. Treatment with TGF-beta1 or activin A down-regulated E cadherin in B16 cells in a dose-dependent manner. In epithelial cells, TGF-beta induced high mobility group A2 (HMGA2) gene product is suggested to down-regulate E-cadherin through up-regulation of zinc-finger transcription factors Slug and Snail, and basic helix-loop-helix transcription factor Twist. Unlike the regulation in epithelial cells, TGF-beta1 treatment rather decreased mRNA expression of HMGA2, indicating a distinct mechanism on TGF-beta/activin-induced down-regulation. Transfection of double-stranded interfering RNA (dsRNAi) for activin receptor-like kinase (ALK) type I receptors revealed that ALK5, a prototype of TGF-beta receptor, mainly transmits TGF-beta signals on the E cadherin down-regulation at the mRNA level, and that a prototype receptor ALK4 elicited the activin effect. TGF-beta/activin potentiated down-regulation of E cadherin and HMGA2 also in B16 sublines that are susceptible to metastasis. However, the extent of down-regulation tended to be smaller, and less Smad2, a signal mediator for TGF-beta/activin, was phosphorylated in response to the ligand, resulting from less expression of type I receptors in the B16 sublines. These results suggest that the receptor expression level determines strength of the signals for TGF-beta/activin through phosphorylation of Smad2, which explains pluripotency of the ligand family partly. PMID- 19288219 TI - Overcoming drug resistance in hormone- and drug-refractory prostate cancer cell line, PC-3 by docetaxel and gossypol combination. AB - Drug resistance is a significant challenge of daily oncology practice. Docetaxel and gossypol both have antitumoral activity in hormone-refractory prostate cancer (HRPC). Our results revealed that docetaxel and gossypol were synergistically cytotoxic and apoptotic in PC-3 cells in a dose- and time-dependent manner. We further investigated the expression profiles of genes involved in drug resistance and metabolism with a Human Cancer Drug Resistance and Metabolism PCR Array (SuperArray). Six of the 84 genes that are known to regulate drug resistance, metabolism, cell cycle, DNA repair and oncogenesis were downregulated >or=3-fold change by the combination treatment. These results may be important in devising mechanism-based and targeted therapeutic strategies for prostate cancer, especially in devising combination therapy for drug resistant prostate cancers. PMID- 19288220 TI - Isolation and characterization of a cotton cdh-like gene. AB - Cotton fiber cells elongate without dividing to form economically valuable spinnable fiber. Reports of the ploidy level of fiber cells are variable. Early reports indicated an increase in nuclear DNA content in young fibers; however, subsequent reports failed to observe such a significant increase in ploidy level. Evaluation and analysis of genes involved in regulation of DNA synthesis and other aspects of cell cycle regulation identified relevant genes that were present in fiber cells though usually at low levels. We report the isolation and characterization of another gene likely to be involved in cell cycle/DNA synthesis control. This gene was similar to a gene from Medicago species that controls entry into anaphase by regulating the activity of the anaphase promoting complex ability to ubiquinate selected proteins. The cotton gene was composed of nine exons and the deduced translational sequences have motifs similar to a Medicago gene expressed in highly polyploid cells. Based on this similarity the cotton gene was designated Ghcdh. Ghcdh is highly expressed in meristems and leaves but is present at much lower levels in fiber cells. These data are consistent with the lower levels of polyploidy reported for cotton fiber. A simple sequence repeat was identified in the gene that may be exploited as a marker to map this gene and associate it with important traits in cotton. PMID- 19288221 TI - Identification of low abundance polyA-binding proteins in Arabidopsis chloroplast using polyA-affinity column. AB - Proteins could be well separated and further identified by the use of 2-DE and related techniques. Yet, there are many proteins could not be detected even by more effective dyes because of their inherent low abundance or their low resolution. As a result, polyA-affinity column was used as a method to enrich polyA-binding proteins and then identified by MALDI-TOF-MS. In this study, 23 Arabidopsis chloroplast protein spots coded by 18 genes were identified, and majority of these proteins were classified into three related categories according to their annotations in the Swiss-Prot database, including NAD-, RNA-, and ATP-binding motifs, respectively. The major goal of the present Arabidopsis chloroplast proteomics project was to identify novel polyA-binding proteins or protein isoforms located in Arabidopsis chloroplasts and the specific research of cellular proteins with extremely low transcription levels could be fulfilled. PMID- 19288222 TI - Replenishing a computerized adaptive test of patient-reported daily activity functioning. AB - PURPOSE: Computerized adaptive testing (CAT) item banks may need to be updated, but before new items can be added, they must be linked to the previous CAT. The purpose of this study was to evaluate 41 pretest items prior to including them into an operational CAT. METHODS: We recruited 6,882 patients with spine, lower extremity, upper extremity, and nonorthopedic impairments who received outpatient rehabilitation in one of 147 clinics across 13 states of the USA. Forty-one new Daily Activity (DA) items were administered along with the Activity Measure for Post-Acute Care Daily Activity CAT (DA-CAT-1) in five separate waves. We compared the scoring consistency with the full item bank, test information function (TIF), person standard errors (SEs), and content range of the DA-CAT-1 to the new CAT (DA-CAT-2) with the pretest items by real data simulations. RESULTS: We retained 29 of the 41 pretest items. Scores from the DA-CAT-2 were more consistent (ICC = 0.90 versus 0.96) than DA-CAT-1 when compared with the full item bank. TIF and person SEs were improved for persons with higher levels of DA functioning, and ceiling effects were reduced from 16.1% to 6.1%. CONCLUSIONS: Item response theory and online calibration methods were valuable in improving the DA-CAT. PMID- 19288223 TI - Association of insomnia with quality of life, work productivity, and activity impairment. AB - PURPOSE: To assess the association of insomnia with health-related quality of life (HRQOL), work productivity, and activity impairment. METHODS: Data were obtained from the 2005 US National Health and Wellness Survey. Subjects were assigned to the insomnia group (diagnosed insomnia experienced at least a few times a month) or the noninsomnia group (no insomnia or sleep symptoms). HRQOL was assessed using the short form 8 (SF-8) (mental and physical scores). The work productivity and activity impairment questionnaire (WPAI) assessed absenteeism (work time missed), presenteeism (impairment at work), work productivity loss (overall work impairment), and activity impairment. Linear regression models were used to control for potential confounders. RESULTS: A total of 19,711 adults were evaluated (5,161 insomnia, 14,550 noninsomnia). Subjects in the insomnia group had significantly lower SF-8 physical (-5.40) and mental (-4.39) scores and greater activity impairment scores (+18.04) than subjects in the noninsomnia group (P < 0.01 for all). Employed subjects in the insomnia group had greater absenteeism (+6.27), presenteeism (+13.20), and work productivity loss (+10.33) scores than those in the noninsomnia group (P < 0.01 for all). CONCLUSIONS: Insomnia is significantly associated with poorer physical and mental quality of life and work productivity loss and activity impairment. PMID- 19288224 TI - Establishing construct validity for the thyroid-specific patient reported outcome measure (ThyPRO): an initial examination. AB - OBJECTIVE: To establish a reliable and valid scale structure of a patient reported outcome measuring thyroid-specific quality of life. METHODS: The 98-item ThyPRO questionnaire was administered to patients with benign thyroid diseases at two university hospitals. Multi-trait scaling was performed, evaluating lack of convergent validity (item-own scale polyserial correlation <0.40) or lack of discriminant validity (item-other scale correlation higher than item-own scale correlation) of the hypothesized scale structure. Analyses were repeated in clinical and sociodemographic subgroups and with Pearson correlations. Reliability was estimated by Cronbach's alpha, both conventionally and with polychoric correlations. RESULTS: In total, 904 patients (69%) responded. Initial multitrait scaling analysis identified 25 scaling errors. Twelve items were omitted from the scale structure, and a re-analysis showed complete convergent validity and only two instances of lack of discriminant validity. Pearson correlations yielded similar results. Across all subgroups, convergent validity was complete, and discriminant validity was found in 99.2% of tests. Lack of discriminant validity was mainly between physical symptoms and psychological and disease-impact scales. Cronbach's alpha was acceptable (>0.70, >0.80 with polychoric correlations) for all 13 scales. CONCLUSION: A reliable scale structure displaying complete convergent and almost complete discriminant validity was established in general analyses and in distinct clinical subgroups of patients with benign thyroid diseases. PMID- 19288225 TI - Nicotinamide prevents NAD+ depletion and protects neurons against excitotoxicity and cerebral ischemia: NAD+ consumption by SIRT1 may endanger energetically compromised neurons. AB - Neurons require large amounts of energy to support their survival and function, and are therefore susceptible to excitotoxicity, a form of cell death involving bioenergetic stress that may occur in several neurological disorders including stroke and Alzheimer's disease. Here we studied the roles of NAD(+) bioenergetic state, and the NAD(+)-dependent enzymes SIRT1 and PARP-1, in excitotoxic neuronal death in cultured neurons and in a mouse model of focal ischemic stroke. Excitotoxic activation of NMDA receptors induced a rapid decrease of cellular NAD(P)H levels and mitochondrial membrane potential. Decreased NAD(+) levels and poly (ADP-ribose) polymer (PAR) accumulation in nuclei were relatively early events (<4 h) that preceded the appearance of propidium iodide- and TUNEL positive cells (markers of necrotic cell death and DNA strand breakage, respectively) which became evident by 6 h. Nicotinamide, an NAD(+) precursor and an inhibitor of SIRT1 and PARP1, inhibited SIRT1 deacetylase activity without affecting SIRT1 protein levels. NAD(+) levels were preserved and PAR accumulation and neuronal death induced by excitotoxic insults were attenuated in nicotinamide treated cells. Treatment of neurons with the SIRT1 activator resveratrol did not protect them from glutamate/NMDA-induced NAD(+) depletion and death. In a mouse model of focal cerebral ischemic stroke, NAD(+) levels were decreased in both the contralateral and ipsilateral cortex 6 h after the onset of ischemia. Stroke resulted in dynamic changes of SIRT1 protein and activity levels which varied among brain regions. Administration of nicotinamide (200 mg/kg, i.p.) up to 1 h after the onset of ischemia elevated brain NAD(+) levels and reduced ischemic infarct size. Our findings demonstrate that the NAD(+) bioenergetic state is critical in determining whether neurons live or die in excitotoxic and ischemic conditions, and suggest a potential therapeutic benefit in stroke of agents that preserve cellular NAD(+) levels. Our data further suggest that, SIRT1 is linked to bioenergetic state and stress responses in neurons, and that under conditions of reduced cellular energy levels SIRT1 enzyme activity may consume sufficient NAD(+) to nullify any cell survival-promoting effects of its deacetylase action on protein substrates. PMID- 19288226 TI - Circulating adiponectin represents a biomarker of the association between adiposity and bone mineral density. AB - An association exists between adiposity, insulin resistance, and osteoporosis; however, the mechanism of this relationship remains enigmatic. We aimed to determine whether the insulin resistance index (HOMA-IR), serum adiponectin, or leptin levels are associated with bone mineral density (BMD). A cross-sectional, observational study was designed. Eighty-four postmenopausal ambulant women [52.5 (50.0-58.0) years; body mass index (BMI): 29.4 (25.9-33.8) kg/m(2)] referred for osteoporosis screening were enrolled. Anthropometric measures, fasting serum adiponectin and leptin levels, and the HOMA-IR were determined. The relationships between these variables and lumbar, hip, and forearm BMD measured by dual-energy X-ray absorptiometry (DXA) were analyzed. Considering all 84 participants, the HOMA-IR index was 1.82 (1.17-2.86), serum adiponectin was 13.25 (10.49-16.88) microg/ml, and serum leptin was 19.26 (14.94-24.90) ng/ml. BMI, waist circumference, and leptin positively correlated with hip and lumbar BMD, whereas adiponectin negatively correlated. Multivariate analysis confirmed an inverse relation between serum adiponectin level and femoral neck and lumbar BMD measurements. In total hip and forearm areas, there was no independent association of adipocytokines with BMD measurements. Instead, waist circumference was independently associated with BMD measurements. In conclusion, adiponectin may represent a biomarker in the relationship between visceral fat mass and BMD. However, this association is probably confounded by the specific body composition parameters (i.e., waist circumference, BMI) in postmenopausal women. PMID- 19288227 TI - Expression of microRNAs in cotton. AB - MicroRNAs (miRNAs) are a class of newly identified non-protein-coding small RNAs. miRNAs post-transcriptionally regulate the expression of more than 30% genes, which control many biological and metabolic progresses, including plant growth, development, and response to environmental stresses. However, no studies have been performed on miRNA expression in cotton, one of the most important fiber and economic crops. In this study, we employed quantitative real-time PCR (qRT-PCR) to detect and compare miRNA expressions in eight different cotton organs at different developmental stages. Our results showed that miRNAs were differentially expressed in different cotton organs, with certain classes expressed preferentially in an organ-specific manner. The miR-156 was highly expressed in cotyledon, whereas the miR-172 was highly expressed in young leaves at fruit branch, young flower buds, 0 day post-anthesis (DPA) ovules, and 0 DPA petals. We also found that miR-172 was not highly expressed in all parts of flowers. In contrast, miR-172 was highly expressed in petal but not in stamen and carpel. Interestingly, miR-162 was highly expressed in immature fiber, 2 DPA ovules, and mixtures of 0 DPA stamen and carpel, suggesting miR-162 may play a role in cotton fiber differentiation and development. Our previous study showed that miR-396 may target a fiber-related gene the callose synthase catalytic subunit. In this study, the miR-396 expression was observed in all eight cotton organs. This study identified the expression of miRNAs that may regulate the development of cotton and cotton fiber. PMID- 19288228 TI - Genetic diversity of Hevea IRRDB'81 collection assessed by RAPD markers. AB - The majority of Hevea (Hevea brasiliensis Muell. Arg.) genetic resource in Vietnam derived from the IRRDB'81 germplasm collected in the Amazonian habitats of the genus. Random amplified polymorphic DNA (RAPD) analysis was used to examine the genetic diversity and structure of the IRRDB'81 germplasm. A total of 59 accessions from 13 different districts of the Brazilian states namely Acre, Rondonia, and Mato Grosso were brought into the study using six arbitrarily preselected primers. Sixty-five RAPD band patterns ranging in size from 0.2 to 3.0 kbp were scored for analysis. Differences in the level of DNA polymorphism among the districts and states were revealed. The percentage of the polymorphic DNA fragments calculated for 13 individual districts varied from 15.38 to 70.77%. The mean values of heterozygosity within the district varied from 0.064 to 0.264. Pairwise district Nei's genetic distance values ranged from 0.046 for Catriquacu and Itanba of Mato Grosso to 0.304 for Tarauaca of Acre and Aracatuba of Mato Grosso. The estimated values of Shannon's diversity index ranged from 0.093 for the Assis-Brasil district of Acre to 0.389 for the Jiparana district of Rondonia. The analysis of molecular variance (AMOVA) indicated that most of the genetic variations were found among accessions within the districts, while interdistrict variance component accounted for 14.1% only. The low interdistrict differentiation probably implied an extensive gene flow among them. Both the principal coordinate analysis and UPGMA cluster analysis based on genetic distance values revealed a varying degree of separation among the districts and that conformed to geographical origins of Hevea IRRDB'81 collection. PMID- 19288229 TI - Genomic subtraction recovers rye-specific DNA elements enriched in the rye genome. AB - Repetitive DNA sequence families have been identified in methylated relic DNAs of rye. This study sought to isolate rye genome-specific repetitive elements regardless of the level of methylation, using a genomic subtraction method. The total genomic DNAs of rye-chromosome-addition-wheat lines were cleaved to short fragments with a methylation-insensitive 4-bp cutter, MboI, and then common DNA sequences between rye and wheat were subtracted by annealing with excess wheat genomic DNA. Four classes of rye-specific repetitive elements were successfully isolated from both the methylated and non-methylated regions of the genome. Annealing of the DNA mixture at a ratio of the enzyme-restricted fragments:the sonicated fragments (1:3-1:5) was key to this success. Two classes of repetitive elements identified here belong to representative repetitive families: the tandem 350-family and the dispersed R173 family. Southern blot hybridization patterns of the two repetitive elements showed distinct fragments in methylation-insensitive EcoO109I digests, but continuous smear signals in the methylation-sensitive PstI and SalI digests, indicating that both of the known families are contained in the methylated regions. The subtelomeric tandem 350-family is organized by multimers of a 380-bp-core unit defined by the restriction enzyme EcoO109I. The other two repetitive element classes had new DNA sequences (444, 89 bp) and different core unit sizes, as defined by methylation-sensitive enzymes. The EcoO109I recognition sites consisting of PyCCNGGPu-multi sequences existed with high frequency in the four types of rye repetitive families and might be a useful tool for studying the genomic organization and differentiation of this species. PMID- 19288230 TI - Single molecule detection in nanofluidic digital array enables accurate measurement of DNA copy number. AB - Digital polymerase chain reaction (PCR) is a promising technique for estimating target DNA copy number. PCR solution is distributed throughout numerous partitions, and following amplification, target DNA copy number is estimated based on the proportion of partitions containing amplified DNA. Here, we identify approaches for obtaining reliable digital PCR data. Single molecule amplification efficiency was significantly improved following fragmentation of total DNA and bias in copy number estimates reduced by analysis of short intact target DNA fragments. Random and independent distribution of target DNA molecules throughout partitions, which is critical to accurate digital PCR measurement, was demonstrated by spatial distribution analysis. The estimated relative uncertainty for target DNA concentration was under 6% when analyzing five digital panels comprising 765 partitions each, provided the panels contained an average of 212 to 3,365 template molecules. Partition volume was a major component of this uncertainty estimate. These findings can be applied to other digital PCR studies to improve confidence in such measurements. PMID- 19288231 TI - A light upon the horizon: young analytical chemists forging new scientific pathways during an economic downturn. PMID- 19288232 TI - Using the Tsallis distribution and the fractional differentiation to resolve the overlapping bands. AB - Using the Tsallis distribution, which facilitates the generalization of well known distributions such as Gaussian and Lorentzian by varying a non-extensivity parameter q as a model of the individual band to correctly assign overlapping bands and the fractional differentiation as mathematical tool to help to determine the spectral parameters of the individual band, a new resolution method for the overlapping bands is presented. According to variation of the maximum and the zero-crossing of the Tsallis distribution at different differential order, two types of parameter estimators are obtained, which are utilized to calculate the parameters of position, height, and width of Tsallis distribution. To verify the suggested method, separation of several kinds of overlapping bands simulated by computer and the experimental infrared spectrum of 1,2-bromofluoroethane have been performed and discussed. PMID- 19288233 TI - Exposure to lead and mercury in young larvae induces more severe deficits in neuronal survival and synaptic function than in adult nematodes. AB - In the present study, we investigated the possibly neurotoxic effects of metal (Pb and Hg) exposure at different developmental stages on neuronal loss in the GABAergic nervous system and synaptic functions in the nematode Caenorhabditis elegans. Our data suggest that neuronal survival in GABAergic neurons and cholinergic transmission were relatively stable during development in nematodes. Moreover, neurodegeneration, as shown by the neuronal loss and dorsal/ventral cord gaps, was more severely induced by Pb and Hg exposure at the L1 through L3 larval stages than at the L4 larval and young-adult stages. Similarly, pre- and postsynaptic functions were more severely impaired by Pb and Hg exposure at the L1 through L3 larval stages than at the L4 larval and young-adult stages. Furthermore, both aldicarb and levamisole resistance were significantly correlated with neuronal loss, dorsal cord gap, and ventral cord gap in Pb- and Hg-exposed nematodes, suggesting that neuronal survival was noticeably correlated with synaptic function in metal-exposed nematodes during development. Therefore, younger (L1-L3) larvae show more sensitivity to neurotoxicity of neuronal survival and synaptic function than L4 larvae and young adult nematodes. PMID- 19288234 TI - Prevalence of Helicobacter pylori seropositivity among patients undergoing bariatric surgery: a preliminary study. PMID- 19288235 TI - The year in Clinical Research in Cardiology. PMID- 19288236 TI - Nucleotide sequence and genome organization of the medium RNA of Iris yellow spot virus from the United States. AB - Iris yellow spot tospovirus (IYSV) of the family Bunyaviridae causes a serious disease in onion in the USA and other parts of the world. Inspite of its economic importance, the complete genomic sequence of IYSV from the USA is not available. The genome structure and organization of the medium (M) RNA of a Washington (WA) isolate of IYSV were determined and compared to the corresponding region of two isolates previously described from Brazil and The Netherlands. Sequence analysis showed that the M-RNA was 4,817 nucleotides long and potentially coded for the movement protein (NSm) in the viral sense and the glycoprotein precursor (Gn and Gc) in the viral complementary sense. The predicted sizes of NSm and Gn/Gc precursor were 34.7 and 128.84 kDa, respectively. The two open reading frames are separated by a 380 nucleotide intergenic region. Phylogenetic analysis of the NSm and Gn/Gc genes from the WA isolate showed grouping that reflected their respective serogroups. The WA isolate formed a close cluster with the two previously reported IYSV isolates and the IYSV cluster was distinguishable from other tospovirus species. This is the first report of complete genomic sequence of the M-RNA of IYSV from the US. PMID- 19288237 TI - Lipid rafts play an important role in the vesicular stomatitis virus life cycle. AB - Lipid rafts are involved in the life cycle of many viruses. In this study, we investigated the role of lipids in the life cycle of vesicular stomatitis virus (VSV). Cholesterol depletion by pretreatment of BHK cells or VSV particles with methyl-beta-cyclodextrin (MbetaCD), a cholesterol-sequestering drug, inhibited the production of VSV dramatically. This effect was reversible, and virus production was restored by the addition of cholesterol, indicating that the reduction was caused by the loss of cholesterol in the cell membrane and virus, respectively. Cholesterol depletion at the adsorption stage also reduced the production of VSV significantly, but in contrast, only had a limited effect on virus production at the post-entry stage. Inhibition of sphingomyelin by myriocin treatment only showed a minor effect on VSV production. However, reduction of cholesterol and sphingomyelin at the same time dramatically reduced VSV production, showed a significant synergistic effect. These results suggest that lipid rafts play an important role in the life cycle of VSV. PMID- 19288238 TI - Retroperitoneal sepsis with mediastinal and subcutaneous emphysema complicating stapled transanal rectal resection (STARR). AB - The STARR procedure was introduced a few years ago for the treatment of obstructed defaecation syndrome secondary to internal rectal intussusception and rectocele. We present a case of severe retroperitoneal sepsis with mediastinal and subcutaneous emphysema complicating STARR, treated by transperineal pelvic drainage and a loop sigmoid colostomy. PMID- 19288239 TI - Locally advanced anourological verrucous carcinoma: a multidisciplinary approach. PMID- 19288240 TI - A simple technique to control faecal spillage during proctoscopic detorsion of sigmoid volvulus. PMID- 19288241 TI - Ultra-low anterior resection for low rectal cancer: five key tips to make it easy. AB - The primary treatment for rectal cancer is still surgery. Surgery however, may be either preceded or followed by chemotherapy and radiotherapy as and when needed. Good surgery on its own when applied appropriately is associated with a very low rate of local recurrence. Bad surgery however, is associated with a high risk of local and systemic recurrence. The well-performed ultra-low anterior resection with total mesorectal clearance has been shown to be the most important step in the treatment of a low rectal cancer. The author details five key tips to make the performance of an ultra-low anterior resection easy. These five key tips are: tip 1 Decide on the best approach for your particular patient before surgery; tip 2 The inferior mesenteric artery is crucial in obtaining the right dissection plane; tip 3 The sigmoid colon should be used for construction of the J-pouch whenever possible; tip 4 Use of a good light source is the key to obtaining a good idea of where to operate and to keeping to the right planes; tip 5 The last key to making surgery easier is to have a good head and good hands. Surgeons must know precisely the reasons why they do what they do in the way they do. Surgery which is difficult may be expected to carry with it more risks and therefore poorer results. Surgery which goes smoothly and is easy is usually rewarded with good results. Surgeons should try to make their surgery easier and the tips offered here are steps that will do this. PMID- 19288242 TI - Laparoscopic-assisted sigmoidectomy with transanal specimen extraction: a bridge to NOTES? AB - The combination of standard laparoscopy and specimen extraction through a natural orifice has the potential to decrease wound related complications. We describe the technical approach to laparoscopic sigmoidectomy in which the specimen is extracted transanally through a proctoscope. Laparoscopic-assisted sigmoidectomy with transanal specimen removal avoids an abdominal wall extraction incision and may be considered an alternative approach to conventional minimally invasive sigmoidectomy in patients with disease of the sigmoid or left colon. PMID- 19288243 TI - Laparoscopic high anterior resection with natural orifice specimen extraction (NOSE) for early rectal cancer. AB - Laparoscopic surgery for colorectal cancer requires an abdominal incision to extract the resected specimen. We describe a technique for laparoscopic resection of an early-stage upper rectal cancer in a 51-year-old man followed by transanal specimen delivery, hence avoiding the need for making any additional abdominal incisions for retrieval of the specimen. Pneumoperitoneum was created, followed by medial-tolateral mobilization of the sigmoid colon, and take down of the splenic flexure and division of the inferior mesenteric vessels laparoscopically. The upper rectum distal to the tumour and proximal colon was transected with a laparoscopic stapler. The specimen was retrieved transanally via an opening in the rectal stump. The proximal colon was then delivered transanally and the anvil of the circular stapler inserted before returning it to the pelvic cavity. The rectal stump was transected again just below the opening to close off the stump, and the colorectal anastomosis was then completed intracorporeally. The patient, a 51-year-old male (BMI 18.6 kg/m(2)) with a 2.5-cm, early-stage posterior rectal cancer 12 cm from the anal verge, underwent the above-described procedure. Postoperative recovery was uneventful. He resumed normal daily activities 1 week after surgery. Histology confirmed a T1N0 upper rectal cancer. In the effort to minimize surgical trauma and postoperative pain, natural orifice specimen extraction techniques have been attempted. This procedure may be applicable to benign tumours and early colorectal cancer, and serves as an intermediate step between laparoscopic and natural orifice surgery. PMID- 19288244 TI - Anorectal transplantation. AB - BACKGROUND: Anorectal transplantation is a valid procedure for the treatment of anorectal dysfunction; however, the lack of a suitable animal model has hampered the development of this method. We describe a simple technique for anorectal transplantation in the rat and compare this procedure with colostomy. METHODS: The anorectal segment including the skin surrounding the anus were freed by abdominal and perineal dissection. In a heterotopically transplanted group the segment was exteriorized by the formation of an anus through an abdominal incision. In an orthotopically transplanted group the segment was replaced in its original position and reimplanted by suturing. In another group a distal colostomy was performed. A sham-treated control group (simulated surgical procedure) was also included. Changes in behavior, characteristics of the stool, body weight and survival rate were assessed by daily clinical examination. Moribund animals, those with a weight loss of more than 30%, and those surviving at 1 month were killed by an overdose of anesthetic. The results were analyzed using the Mann Whitney, Student's t and chi-squared tests, and p<0.05 was considered significant. RESULTS: Within 4 days after the operation, animals submitted to orthotopic or heterotopic transplantation had achieved normal defecation, body weight gain and clinical evolution similar to the sham-treated group. The overall mortality in these groups was 4.16%. In contrast, colostomized animals showed a high incidence of diarrhea, intestinal obstruction, stress posture and violent behavior (p0.05). Recurrence rates were significantly lower in the SM(mod) group (2/19, 10.5%) than in the SM(std) group (14/21, 66.7%; p=0.0004). The percentage of patients highly satisfied (score -1) with the procedure was significantly higher in the SM(mod) group (73.7%, 14/19) than in the SM(std) group (33.3%, 7/21; p=0.014). CONCLUSION: Patients with high-grade haemorrhoids in whom a modified stapled mucosectomy (ligating and cutting the residual haemorrhoids on the operating table after the procedure) was performed had a significantly lower recurrence rate and better satisfaction rate than patients in whom standard stapled mucosectomy was performed. Long-term studies in a larger group of patients are needed to substantiate these results. PMID- 19288252 TI - NOTES/NOSE/NOSCAR/LATAS: what does it all mean? PMID- 19288253 TI - Histone H3 acetylation of StAR and decrease in DAX-1 is involved in the luteinization of bovine granulosa cells during in vitro culture. AB - We investigated the expression of genes and transcription factors associated with steroidogenesis during the luteinization of granulosa cells isolated from bovine small follicles. Granulosa cells produced progesterone when cultivated in a culture medium including serum and attached to the substrate and began to display an elongated or fibroblastic aspect within 24 h of culture. We observed an increase in the number of granulosa cells at the same time. P450arom expression in the cultured granulosa cells had decreased at 24 h compared with 0 h of culture, and afterward was maintained at a low level. This expression was consistent with the decline of E2 concentration in the medium. Expression of StAR and P450scc mRNAs in the cultured granulosa cells was significantly increased at 72 h compared with 0 h of culture. Although the expression of Ad4BP/SF-1 mRNA began to increase during period between 48 and 72 h of culture, protein expression of Ad4BP/SF-1 remained at a constant level throughout the culture period. DAX-1 mRNA expression had decreased at 24 h of culture and remained at a low level. In parallel with this expression, the protein expression of DAX-1 began to decrease between 24 and 48 h of culture and then remained at a low level. Histone H3 acetylation of the StAR promoter region was observed at 72 h of culture period. Our data suggested that the decrease of Dax-1 transcription factor and the increase in histone H3 acetylation may play important roles in progesterone synthesis in luteinizing granulosa cells. PMID- 19288254 TI - The heritability of aptitude and exceptional talent across different domains in adolescents and young adults. AB - The origin of individual differences in aptitude, defined as a domain-specific skill within the normal ability range, and talent, defined as a domain specific skill of exceptional quality, is under debate. The nature of the variation in aptitudes and exceptional talents across different domains was investigated in a population based twin sample. Self-report data from 1,685 twin pairs (12-24 years) were analyzed for Music, Arts, Writing, Language, Chess, Mathematics, Sports, Memory, and Knowledge. The influence of shared environment was small for both aptitude and talent. Additive and non-additive genetic effects explained the major part of the substantial familial clustering in the aptitude measures with heritability estimates ranging between .32 and .71. Heritability estimates for talents were higher and ranged between .50 and .92. In general, the genetic architecture for aptitude and talent was similar in men and women. Genetic factors contribute to a large extent to variation in aptitude and talent across different domains of intellectual, creative, and sports abilities. PMID- 19288255 TI - Effects of repeated crowding on the stress response and growth performance in Atlantic salmon (Salmo salar). AB - A 64-day growth experiment was conducted in which two groups of Atlantic salmon parr were grown under either control conditions or subjected to a weekly crowding stressor. Subjecting fish to the stressor resulted in a 7.7% reduction in wet weight after 29 days, which was maintained at 7.9% by day 64. This reduction in weight was reflected in a 44% reduction in specific growth rate and 38% increase in feed conversion ratio over the first 29 days of the experiment. Elevation in plasma cortisol was observed in crowded fish on days 1, 29 and 64. Similarly, on days 1 and 29 an increase in both plasma glucose and lactate was detected. On day 64, however, no differences in plasma glucose and lactate were observed, with the magnitude of the cortisol response also significantly reduced. Overall, the relatively moderate impact on growth performance and reduction in magnitude of measured stress parameters at the end of the experiment suggests possible habituation to the applied stressor. PMID- 19288256 TI - Comparative fatty acid profiles of wild and farmed tropical freshwater fish rohu (Labeo rohita). AB - The proximate composition of the whole body and the fatty acid composition of the liver, muscle, eye and brain of wild and cultured rohu (Labeo rohita) were analyzed. The cultured species was found to have significantly (P < 0.05) higher lipid contents than its wild counterpart. The saturated (SFA) and monounsaturated (MUFA) fatty acid contents were significantly higher in the cultured species, whereas the n-6 and n-3 polyunsaturated fatty acid (PUFA) levels were higher in the wild species. Fatty acids C16:0 and C18:1 n-9 were the principal fatty acids of the SFAs and MUFAs, respectively, identified in the analyses. Docosahexaenoic acid, eicosapentaenoic acid, and arachidonic acid were the predominant PUFAs in both groups, and all three were found to be present at significantly (P < 0.05) higher levels in the wild species. Erucic acid (C22:1 n-9), which was the predominant fatty acid (30.76%) in the feed, was detected only at low levels in muscle (0.30%), liver (1.04%) and eye (1.28%) of cultured fish tissue. PMID- 19288257 TI - Development of visual cells in the Pacific bluefin tuna Thunnus orientalis. AB - The development of rod and cone photoreceptor cells was investigated in the retinas of Pacific bluefin tuna larvae and juveniles, using RET-P1 monoclonal antibody labeling to identify photoreceptors. At 60 h after hatching, which was about when feeding began, opsin (presumably green opsin (Rh2)) was expressed in the outer segments of cone cells. At 15 days after hatching (dah), although many labeled cone cells were observed in the dorsal retina, the same type of cone cells had partially appeared in the ventral retina. The presence of rod cell bodies was confirmed by the expression of Rh1 opsin at 15 dah. At 21 dah, the presence of outer segments of rod cells was confirmed by the expression of Rh1 opsin and by morphology. The observations suggest that the cone cells were substantially operable upon the development of their outer segment at around the beginning of the post-larval stage, and the rod cells began to function at around 15 to 21 dah, before and during metamorphosis. PMID- 19288258 TI - Experience is a good, but not perfect, teacher. PMID- 19288259 TI - Contrast-enhanced cardiovascular magnetic resonance in the hyperacute phase of ST elevation myocardial infarction. AB - Cardiovascular magnetic resonance (CMR) very early after primary percutaneous coronary intervention (PPCI) may lead to instability or early stent complications. However, CMR in the hyperacute phase of STEMI may improve risk stratification. We investigated feasibility and safety of CMR in the hyperacute phase of STEMI immediately after PPCI. One hundred and twenty eight consecutive patients immediately after PPCI for STEMI. Sixty four underwent CMR <12 h after PPCI versus 64 matched controls. Outcomes were followed over 6 months. CMR in hyperacute STEMI was not associated with in-hospital death, infarct expansion, or urgent revascularization (P = NS). CMR (32 ml gadolinium contrast) immediately after PPCI (180 ml iodine contrast) did not increase nephropathy. CMR did not increase major adverse cardiac events (5 vs. 8%, P = 0.16) or recurrence of angina (6 vs. 8%, P = 0.73) at 6 months. CMR immediately after PPCI is feasible and safe, allowing very early risk stratification in STEMI. PMID- 19288260 TI - Drug evaluation: vildagliptin-metformin single-tablet combination. AB - The single-tablet combination of vildagliptin and metformin addresses key defects of type 2 diabetes for improved glycemic control. By inhibiting the dipeptidyl peptidase-4 (DPP-4) enzyme, vildagliptin raises the levels of the active incretin hormones, glucagon-like peptide 1 and glucose-dependent insulinotropic peptide. This leads to increased synthesis and release of insulin from the pancreatic beta cells and decreased release of glucagon from the pancreatic alpha cells. The combination tablet also contains metformin, which addresses insulin resistance. The complementary mechanisms of action of the two agents in combination have been shown to provide additive and sustained reductions in hemoglobin A(1c) compared with metformin monotherapy. In active-controlled trials, the vildagliptin metformin combination has been shown to produce equivalent reductions in hemoglobin A(1c) to pioglitazone-metformin and glimepiride-metformin combinations, without significant risk of hypoglycemia and without causing weight gain. In clinical trials, the overall incidence of any adverse event was similar in patients randomized to vildagliptin plus metformin and placebo plus metformin. Available data support the use of vildagliptin in combination with metformin as a promising second-line treatment for the management of type 2 diabetes and this is reflected in the latest UK National Institute for Health and Clinical Excellence draft guideline for consultation on new agents for blood glucose control in type 2 diabetes. PMID- 19288261 TI - The glutathione S-transferase inhibitor 6-(7-nitro-2,1,3-benzoxadiazol-4 ylthio)hexanol overcomes the MDR1-P-glycoprotein and MRP1-mediated multidrug resistance in acute myeloid leukemia cells. AB - PURPOSE: There has been an ever growing interest in the search for new anti-tumor compounds that do not interact with MDR1-Pgp and MRP1 drug transporters and so circumvent the effect of these proteins conferring multidrug resistance (MDR) and poor prognosis in AML patients. We have investigated the cytotoxic activity of the strong glutathione S-transferase (GST) inhibitor 6-(7-nitro-2,1,3 benzoxadiazol-4-ylthio)hexanol (NBDHEX) on AML (HL60) cell lines. METHODS: Functional drug efflux studies and cell proliferation assays were performed on both sensitive and MDR AML (HL60) cells after incubation with NBDHEX. Moreover, the mode of cell death (apoptosis vs. necrosis) as well as the correlation between NBDHEX susceptibility and GST activity or Bcl-2 expression was investigated. RESULTS: NBDHEX is not a substrate of either MDR1-Pgp or MRP1 efflux pumps; in fact, it is not only cytotoxic toward the parental HL60 cell line, but also overcomes the MDR phenotype of its HL60/DNR and HL60/ADR variants. CONCLUSIONS: The data herein reported show that NBDHEX mediates efficient killing of both MDR1-Pgp and MRP1 over-expressing AML cells. Therefore, this drug can potentially be used as an effective agent for treating MDR in AML patients. PMID- 19288262 TI - Evolution of age-dependent sex-reversal under adaptive dynamics. AB - We investigate the evolution of the age (or size) at sex-reversal in a model of sequential hermaphroditism, by means of the function-valued adaptive dynamics. The trait is the probability law of the age at sex-reversal considered as a random variable. Our analysis starts with the ecological model which was first introduced and analyzed by Calsina and Ripoll (Math Biosci 208(2), 393-418, 2007). The structure of the population is extended to a genotype class and a new model for an invading/mutant population is introduced. The invasion fitness functional is derived from the ecological setting, and it turns out to be controlled by a formula of Shaw-Mohler type. The problem of finding evolutionarily stable strategies is solved by means of infinite-dimensional linear optimization. We have found that these strategies correspond to sex reversal at a single particular age (or size) even if the set of feasible strategies is considerably broader and allows for a probabilistic sex-reversal. Several examples, including in addition the population-dynamical stability, are illustrated. For a special case, we can show that an unbeatable size at sex reversal must be larger than 69.3% of the expected size at death. PMID- 19288263 TI - Convergence time to the Ewens sampling formula in the infinite alleles Moran model. AB - In this paper, we establish an upper bound for time to convergence to stationarity for the discrete time infinite alleles Moran model. If M is the population size and mu is the mutation rate, this bound gives a cutoff time of log(Mmu)/mu generations. The stationary distribution for this process in the case of sampling without replacement is the Ewens sampling formula. We show that the bound for the total variation distance from the generation t distribution to the Ewens sampling formula is well approximated by one of the extreme value distributions, namely, a standard Gumbel distribution. Beginning with the card shuffling examples of Aldous and Diaconis and extending the ideas of Donnelly and Rodrigues for the two allele model, this model adds to the list of Markov chains that show evidence for the cutoff phenomenon. Because of the broad use of infinite alleles models, this cutoff sets the time scale of applicability for statistical tests based on the Ewens sampling formula and other tests of neutrality in a number of population genetic studies. PMID- 19288264 TI - Serum homocysteine level in patients with ankylosing spondylitis. AB - In this study serum homocystein (Hcy) level was measured and its relationship with disease activity criteria and treatment protocols was investigated in ankylosing spondylitis (AS) patients. Ninety-two AS patients and 58 healthy individuals were recruited. Erythrocyte sedimentation rate and serum C-reactive protein were determined. Bath AS disease activity index and Bath AS functional index were calculated. Serum Hcy levels >15 micromol/l were considered as hyperhomocysteinemia. The mean serum homocysteine levels were 14.40 and 12.60 micromol/l in patients with AS and the control group, respectively, and the difference between two groups was significant. While there was no significant difference between the sulfasalazine (SSZ) group with 14.25 micromol/l mean Hcy level and the methotrexate (MTX)/SSZ group with 16.05 micromol/l, there was a statistically significant difference between the Hcy levels of these two groups and Hcy level of 12.15 micromol/l of the non-steroidal anti-inflammatory drugs group, and 12.60 micromol/l Hcy level of the control group. Mean serum Hcy level was 13.65 micromol/l in patients with active AS and 14.60 micromol/l in patients with inactive AS, and there was no significant difference between the groups. In our study serum Hcy level was found to be significantly higher in patients with AS than in healthy control subjects. Especially for the AS patients receiving MTX and SSZ treatment without folic acid supplementation, addition of folic acid to their therapy may decrease the risk of cardiovascular disease which in turn decreases the mortality in these patients, but further prospective studies are needed for supporting these results. PMID- 19288265 TI - Translocation of cell-penetrating peptides and delivery of their cargoes in triticale microspores. AB - Microspore culture is contributing significantly in the field of plant breeding for crop improvement in general and cereals, in particular. In the present study, we investigated the uptake of fluorescently labeled cell-penetrating peptides (CPP; Tat, Tat(2), M-Tat, peptide vascular endothelial-cadherin, transportan) in the freshly isolated triticale microspores (mid-late uninucleate stage). We demonstrated that Tat (RKKRRQRRR) and Tat(2) (RKKRRQRRRRKKRRQRRR) are able to efficiently transduce GUS enzyme (272 kDa) in its functional form in 5 and 14% of the microspores, respectively, in a noncovalent manner. Pep-1, a synthetic CPP, was able to transduce GUS enzyme in its active form in 31% of the microspores. The effect of various endocytic and macropinocytic inhibitors on Tat(2)-mediated GUS enzyme delivery was studied and revealed a preferred micropinocytosis entry. DNase I protection assay and confocal laser microscopy was carried out to recommend a ratio of 4:1 Tat(2)-linear plasmid DNA (pActGUS) in complex preparation for microspore transfection. We further show that Tat(2) can successfully deliver GUS gene in near to 2% triticale microspores. The negative control mutated Tat (M-Tat: AKKRRQRRR) failed to transducer the GUS protein and transfect the GUS gene in microspore nucleus. The ability of CPPs to deliver macromolecules (protein as well as linear plasmid DNA) noncovalently has been demonstrated in triticale isolated microspores. It further confirms potential applications of CPPs in developing simple, time saving, cost effective plant genetic engineering technologies. PMID- 19288266 TI - Targeted genomic disruption of H-ras and N-ras has no effect on early renal changes after unilateral ureteral ligation. AB - PURPOSE: To assess the contribution of two different Ras monomeric GTPases isoforms H- and N-Ras in the early changes associated to obstructive nephropathy induced by unilateral ureteral obstruction (UUO). METHODS: UUO was performed in N ras (N-ras-/-) and H-ras (H-ras-/-) knock-out mice and control (H-ras+/+/N ras+/+) mice of C57Bl/6 background. Fibronectin, alpha-smooth muscle actin, cleaved caspase-3, ki-67, Ras-GTP, pERK, and pAkt expression was analyzed by western blot and/or immunohistochemistry. Ras isoforms activation and caspase activity were determined by both western blot and ELISA. RESULTS: Three days after UUO, obstructed (O) kidneys of H-ras-/-, N-ras-/-and H-ras+/+/N-ras+/+mice showed no significant differences in activated total ras, pERK1/2, pAkt, total Akt levels, fibronectin, alpha-SMA expression, cell proliferation, and activated caspase-3. The morphological alterations in the O kidneys, revealed by histological and immunohistochemical studies, were also similar in H-ras-/-, N ras-/-, and H-ras+/+/N-ras+/+mice. CONCLUSIONS: These data suggest that the activation of H-ras and N-ras isoforms does not play a major role in the early renal damage induced by UUO. PMID- 19288267 TI - Acute abdomen as initial presentation of gestational choriocarcinoma. AB - Choriocarcinoma is the most malignant tumor of gestational trophoblastic neoplasia. Choriocarcinoma presenting as postpartum hemorrhage and spontaneous uterine perforation with intra-abdominal hemorrhage is very rare. We present a 29 year-old woman with spontaneous uterine rupture due to choriocarcinoma following a live birth pregnancy. The long time interval (2 years) between the previous live birth pregnancy and the diagnosis of the disease, the acute onset of the disease by uterine rupture as the first symptom and the negative urine hCG test are presented and discussed in this case report. PMID- 19288268 TI - Longitudinal analysis of respiratory symptoms in population studies with a focus on dyspnea in marine transportation workers. AB - PURPOSE: Longitudinal respiratory symptoms are rarely studied in occupational epidemiology. We investigated dyspnea change over time and predictors of change over time using two longitudinal modeling techniques, a semi-parametric group based approach (SAS(r) Proc Traj) and a generalized linear mixed model (SAS(r) Proc Glimmix), and compared the two techniques for use in longitudinal studies of respiratory symptoms. METHODS: Data were previously collected from a lung health surveillance study of marine transportation workers. Subjects were seen two to four times over 12 years (1987-1999). At each visit the American Thoracic Society questionnaire was administered and lung function was tested. The semi-parametric group-based model and the generalized linear mixed model were applied to the data. RESULTS: The group-based trajectory model supported two groups of dyspnea change over time. Group 1 (73%) had a steady low-level probability of reporting dyspnea over follow-up, while Group 2 (27%) had an increasing probability of reporting dyspnea over follow-up. The generalized linear mixed model (random intercept) estimated that the probability of reporting dyspnea was increasing over time in the population. Current smoking, female sex, lower lung function and older age were associated with increased probability of reporting dyspnea in both models. CONCLUSIONS: Results from both models indicate that the probability of reporting dyspnea was increasing over time in this occupational cohort. The group based model is capable of identifying multiple patterns of linear and non-linear change while the generalized linear mixed model is preferable when the population mean change (linear) is of interest. Both approaches were able to identify similar characteristics associated with longitudinal dyspnea symptoms. PMID- 19288269 TI - Grass lignin acylation: p-coumaroyl transferase activity and cell wall characteristics of C3 and C4 grasses. AB - Grasses are a predominant source of nutritional energy for livestock systems around the world. Grasses with high lignin content have lower energy conversion efficiencies for production of bioenergy either in the form of ethanol or to milk and meat through ruminants. Grass lignins are uniquely acylated with p-coumarates (pCA), resulting from the incorporation of monolignol p-coumarate conjugates into the growing lignin polymer within the cell wall matrix. The required acyl transferase is a soluble enzyme (p-coumaroyl transferase, pCAT) that utilizes p coumaroyl-CoenzymeA (pCA-CoA) as the activated donor molecule and sinapyl alcohol as the preferred acceptor molecule. Grasses (C3and C4) were evaluated for cell wall characteristics; pCA, lignin, pCAT activity, and neutral sugar composition. All C3 and C4 grasses had measurable pCAT activity, however the pCAT activities did not follow the same pattern as the pCA incorporation into lignin as expected. PMID- 19288270 TI - Substantially reduced expression of PIAS1 is associated with colon cancer development. AB - PURPOSE: Protein inhibitors of activated STATs (PIAS) regulate the interferon gamma (IFN-gamma) signaling pathway, which has numerous effects on tumor development and tumor cell biology. PIAS's also regulate STAT family members not directly involved in IFN-gamma signaling. This project was designed to assess PIAS1 expression in colon cancer. METHODS: To determine whether PIAS1, one of the PIAS family members, or IFN-gamma signaling pathway components could be used to stratify colon tumors, we stained tissue microarrays for PIAS1, interferon regulatory factor-1 (IRF-1) and STAT1alpha. RESULTS: PIAS1 staining of the colon cancer tissue microarrays indicated a strong correlation of normal colon cells, and adenomas, with high expression of both PIAS1 and IRF-1. CONCLUSION: The PIAS1 results in particular may represent a basis for new approaches for efficiently distinguishing adenomas from colon cancer. PMID- 19288271 TI - Interdisciplinary S2k guideline on the diagnosis and treatment of cervical carcinoma. AB - The first treatment recommendations on the diagnosis and treatment of cervical carcinoma were published by experts in the DKG's Working Group on Gynecological Oncology (Arbeitsgemeinschaft Gynakologische Onkologie, AGO) in 1999 (S1 guideline). The purpose of the guidelines is to explicitly present current knowledge regarding specific medical care problems, evaluate them from methodological and clinical viewpoints, clarify contradictory positions, and define the current procedure of choice after weighing up advantages and disadvantages. It is therefore necessary for guidelines to have their validity continually checked and updated. Interdisciplinary collaboration is an indispensable principle for high-quality care in the field of oncology. This should be borne in mind when establishing recommendations for care. The first updating of the guideline was accordingly carried out in May 2004, with a structured consensus process being carried out by an interdisciplinary group of experts representative of the group of users (S2 guideline). The S2 guideline "Diagnosis and Treatment of Cervical Carcinoma" has now been revised again between October 2007 and January 2008 by the AGO's Uterus Commission. PMID- 19288272 TI - Growth of hormone-dependent MCF-7 breast cancer cells is promoted by constitutive caveolin-1 whose expression is lost in an EGF-R-mediated manner during development of tamoxifen resistance. AB - Caveolin-1 displays both tumour-suppressor and tumour-promoter properties in breast cancer. Using characterised preclinical cell models for the transition of oestrogen-sensitive (WT-MCF-7 cells) to a tamoxifen-resistant (TAM-R cells) phenotype we examined the role caveolin-1 in the development of hormone-resistant breast cancer. The WT-MCF-7 cells showed abundant expression of caveolin-1 which potentiated oestrogen-receptor (ERalpha) signalling and promoted cell growth despite caveolin-1 mediating inhibition of ERK signalling. In TAM-R cells caveolin-1 expression was negligible, repressed by EGF-R/ERK signalling. Pharmacological inhibition of EGFR/ERK in TAM-R cells restored caveolin-1 and also resulted in the emergence of pools of phosphorylated caveolin-1. WT-MCF-7 cells exposed to tamoxifen for upto 12 weeks displayed increased caveolin-1 (peaking by week 2) followed (after week 8) by a marked decrease as the cells progress to develop a stable tamoxifen-resistant phenotype. The targeted down regulation (siRNA) of caveolin-1 in WT-MCF-7 cells reduced growth but did not affect their sensitivity to tamoxifen, suggesting loss of caveolin-1 alone is not sufficient to confer tamoxifen-resistance. Hyperactivation of EGFR/ERK is a feature of tamoxifen-resistant breast cancer cells, a principal driver of cell growth. Recombinant expression of caveolin-1 in TAM-R cells did not affect EGFR/ERK activity, potentially due to mislocalisation of caveolin-1 through hyperactivation of the mTOR pathway or altered caveolin-1 phosphorylation. This work defines a novel role for caveolin-1 with implications for the clinical course of breast cancer and identifies caveolin-1 as a potential drug target for the treatment of early oestrogen-dependent breast cancers. Further, the loss of caveolin-1 may have benefit as a molecular signature for tamoxifen resistance. PMID- 19288273 TI - Nerve growth factor of red nucleus involvement in pain induced by spared nerve injury of the rat sciatic nerve. AB - Nerve growth factor (NGF), a member of the neurotrophin family, is essential for the development and maintenance of sensory neurons and for the formation of central pain circuitry. The current study was designed to evaluate the expression of NGF in the brain of rats with spared nerve injury (SNI), using immunohistochemical technique. The results showed that the level of NGF in the Red nucleus (RN) of SNI rats was apparently higher than that of sham-operated rats. To further study the effect of NGF in the development of neuropathic pain, different doses of anti-NGF antibody (20, 2.0 and 0.2 microg/ml) were microinjected into the RN contralateral to the nerve injury side of SNI rats. The data suggested that the higher doses of anti-NGF antibody (20 and 2.0 microg/ml) significantly attenuated the mechanical allodynia of neuropathic rats, while the 0.2 microg/ml antibody showed no analgesic effect. These results suggest that the NGF of RN is involved in the development of neuropathic allodynia in SNI rats. PMID- 19288274 TI - S100B secretion in acute brain slices: modulation by extracellular levels of Ca(2+) and K (+). AB - Hippocampal slices have been widely used to investigate electrophysiological and metabolic neuronal parameters, as well as parameters of astroglial activity including protein phosphorylation and glutamate uptake. S100B is an astroglial derived protein, which extracellularly plays a neurotrophic activity during development and excitotoxic insult. Herein, we characterized S100B secretion in acute hippocampal slices exposed to different concentrations of K(+) and Ca(2+) in the extracellular medium. Absence of Ca(2+) and/or low K(+) (0.2 mM KCl) caused an increase in S100B secretion, possibly by mobilization of internal stores of Ca(2+). In contrast, high K(+) (30 mM KCl) or calcium channel blockers caused a decrease in S100B secretion. This study suggests that exposure of acute hippocampal slices to low- and high-K(+) could be used as an assay to evaluate astrocyte activity by S100B secretion: positively regulated by low K(+) (possibly involving mobilization of internal stores of Ca(2+)) and negatively regulated by high-K(+) (likely secondary to influx of K(+)). PMID- 19288276 TI - Necrostatin-1 reverts shikonin-induced necroptosis to apoptosis. AB - Degterev et al. previously demonstrated that death receptor mediated apoptosis could be diverted to necroptosis when apoptosis signaling was blocked, suggesting that necroptosis may function as a backup mechanism to insure the elimination of damaged cells under certain conditions when apoptosis was inhibited. Here, we show that shikonin-induced necroptosis can be reverted to apoptosis in the presence of necrostatin-1 (Nec-1), a specific necroptosis inhibitor and that the death mode switch is at least partially due to the conversion from mitochondrial inner membrane permeability to mitochondrial outer membrane permeability, which is associated with Bax translocation. The data combined with the previous reports support a notion that apoptosis and necroptosis may function as reciprocal backup mechanisms of cellular demise. To the best of our knowledge, this is the first study to document a conversion from necroptosis to apoptosis. PMID- 19288275 TI - Reduced expression of GABA transporter GAT3 in helpless rats, an animal model of depression. AB - Mood disorders have been linked to glial and synaptic pathology such as disturbed neurotransmission of gamma-aminobutyric acid (GABA). We evaluated the expression of GABAergic marker genes in rats with helpless behaviour, an animal model of depression. Male Sprague-Dawley rats from inbred lines were tested for helpless behaviour and grouped according to failures in terminating foot shock currents. Expression levels of GABAergic marker genes were assessed using semiquantitative in situ-hybridization. Animals with congenital helpless behaviour (cH) were unable to escape current exposure in contrast to cH-animals derived from the same litters with low failure rates and to non-helpless animals (cNH). We found a significant downregulation of the GABA transporter GAT3 in cLH rats. GAT1 showed small changes, glutamic acid decarboxylase (GAD67) and the vesicular GABA transporter were not significantly altered. Reduced GABA transporter expression is well in concert with the behavioural phenotypes of knockout animals and strengthens the hypothesis of impaired glial functions in depression. PMID- 19288277 TI - Antioxidant defense of betaine against isoprenaline-induced myocardial infarction in rats. AB - We investigated the antioxidant preventive effect of betaine on isoprenaline induced myocardial infarction in male albino rats. Isoprenaline induced myocardial infarction was manifested by a moderate elevation in the levels of diagnostic marker enzymes (alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and creatine phosphokinase) and homocysteine in plasma of experimental rats. Significant rise in the level of lipid peroxidation with a concomitant decline in the levels of myocardial non-enzymic (reduced glutathione) and enzymic antioxidants (glutathione peroxidase, glutathione-S-transferase, catalase and superoxide dismutase) was also observed. Oral pretreatment with betaine significantly prevented isoprenaline-induced alterations in the levels of diagnostic marker enzymes and homocysteine in plasma of experimental groups of rats. It counteracted the isoprenaline-induced lipid peroxidation and maintained the myocardial antioxidant defense system at near normal. Histopathological observations also confirmed the protective effect of betaine against isoprenaline induced myocardial infarction. The results of the present investigation indicate that the protective effect of betaine is probably related to its ability to strengthen the myocardial membrane by its membrane stabilizing action or to a counteraction of free radicals by its antioxidant property. PMID- 19288278 TI - Unique opportunities for NMR methods in structural genomics. AB - This Perspective, arising from a workshop held in July 2008 in Buffalo NY, provides an overview of the role NMR has played in the United States Protein Structure Initiative (PSI), and a vision of how NMR will contribute to the forthcoming PSI-Biology program. NMR has contributed in key ways to structure production by the PSI, and new methods have been developed which are impacting the broader protein NMR community. PMID- 19288279 TI - What symptom improvement can be expected after operation for primary hyperparathyroidism? AB - BACKGROUND: The only cure for primary hyperparathyroidism (pHPT) is operative resection of the parathyroid gland(s) responsible for the disease. The 1990 National Institute of Health's (NIH) consensus development conference on asymptomatic pHPT and its subsequent workshop in 2001 established which clinical criteria warranted parathyroidectomy (PTx) versus observation. While there is no debate that these NIH criteria capture a group of patients likely to benefit from PTx, there is concern that these guidelines miss a significant percentage of pHPT patients who actually are symptomatic. Unfortunately, these additional symptoms are often subtle, nonspecific, not traditionally measured, frequently not assessed or considered in this patient population, and are usually attributed to other diagnoses or simply advanced age. METHODS: An evidence-based literature review was performed assessing symptoms and clinical conditions associated with bone, neuropsychiatry, cognition, quality of life, and the neuromuscular system. The level of evidence and grade of recommendations were assigned to key studies to help determine recommendations regarding indications and potential benefits of parathyroidectomy (PTx). RESULTS: Symptoms ranging from decreased bone mineral density, increased fracture risk, neuropsychiatric symptoms and cognitive changes, lower quality of life, and neuromuscular symptoms were found to be associated with pHPT. The effects of PTx on these symptoms was addressed in a variety of studies that varied in quality. CONCLUSION: Although there are few Level I randomized clinical trials addressing the benefit of PTx in patients with pHPT, there are supportive data to suggest that most patients with pHPT would benefit from operative cure. PMID- 19288280 TI - Investigation of the lymphatic stream of the stomach in gastric cancer with solitary lymph node metastasis. AB - BACKGROUND: Understanding the lymphatic drainage route in gastric cancer is crucial for complete lymph node retrieval from sites susceptible to metastasis. However, the lymphatic stream of the stomach is complex and remains incompletely characterized. METHODS: Patients with small (<4 cm) serosa-negative gastric cancer with solitary lymph node metastasis treated at the Cancer Institute Hospital were included in this study. A total of 135 patients were classified according to the location of the solitary lymph node metastasis into the left gastric artery (LGA) group, the right gastroepiploic artery (RGEA) group, the right gastric artery (RGA) group, or the splenic artery (SA) group. The location of the primary tumors was investigated to aid the mapping of the lymphatic stream of the stomach. RESULTS: Lymphatic flow in LGA (65 patients) and in RGEA (57 patients) are main lymphatic drainage routes of the stomach. The lymphatic area overlapped in the lower third of the stomach in LGA and RGEA, and the lymphatic flow associated with gastric cancer located within this overlapped area can be multidirectional. Skip metastases were observed in 13 patients (10%), and all skip metastases were observed in the suprapancreatic area (station 7, 8a, 9, or 11p). CONCLUSIONS: The lymphatic stream of the stomach is complicated and multidirectional. Understanding and mapping the complex lymphatic streams of the stomach will allow surgeons to perform more effective lymph node dissection during gastric cancer surgery. PMID- 19288281 TI - The combined role of ultrasound and frozen section in surgical management of thyroid nodules read as suspicious for papillary thyroid carcinoma on fine needle aspiration biopsy: a retrospective study. AB - BACKGROUND: The purpose of this study was to evaluate the combined role of ultrasound (US) and frozen section (FS) in the surgical management of thyroid nodules that are suspicious for papillary thyroid carcinoma (PTC) on US-guided fine-needle aspiration biopsy (US-FNAB). METHODS: Between 2006 and 2008, 217 patients with thyroid nodules classified as suspicious for PTC on US-FNAB underwent intraoperative FS and surgery. A thyroid nodule with any suspicious US findings of marked hypoechogenicity, microlobulated or irregular margins, microcalcifications, or taller than wider in shape was defined as positive and that without was defined as negative. FSs were classified as malignant, indeterminate, defer, and benign. We compared the results of US groupings, FS, and final histopathologic diagnosis. RESULTS: Twenty-one (52.5%) of 40 patients with a negative US revealed malignancy. In contrast, 168 (94.9%) of 177 patients with a positive US had malignancy. Fourteen (50%) of 28 patients with benign pathology on FS had malignancy, and 166 (97.6%) of 170 patients with malignancy on FS proved malignancy on histopathology. Of 40 patients with negative US, 13 (92.9%) of 14 with malignancy on FS proved malignancy. CONCLUSIONS: The malignancy rate of thyroid nodules read as "suspicious for PTC" on US-FNAB was 87.1%. When a thyroid nodule with "suspicious for PTC" on US-FNAB has suspicious malignant US features, FS may be unnecessary due to a very high risk of malignancy (94.9%). In contrast, when a thyroid nodule read as "suspicious for PTC" on US-FNAB has no suspicious malignant US features, FS can help surgeons determine the extent of surgery. PMID- 19288282 TI - Effects of land mines and unexploded ordnance on the pediatric population and comparison with adults in rural Cambodia. AB - BACKGROUND: This paper was designed to evaluate and compare the impact of explosive war remnants on children versus adults in rural Cambodia. METHODS: A prospective review of trauma database from November 2003 to January 2006 of the Civilian War Victims Surgical Centre in Battambang, Cambodia, run by an Italian NGO called "EMERGENCY" was performed. Age, female ratio, time of evacuation, type of ordnance, pattern of injury, number of operations, transfused patients, hospitalization, mortality, and residual disability were registered and compared. RESULTS: A total of 356 patients acutely wounded by antipersonnel land mines, antitank land mines, or unexploded ordnances (UXO) were admitted. Among these, 94 (26.4%) were children (younger than aged 16 years). Females were more common among children than adults (31.9% vs. 11.8%); 61.7% of children were injured by UXO, whereas 72.1% of adults were victims of antipersonnel land mines. Antitank mines victims were uncommon in both groups. The majority of adults (49.2%) were injured to lower limbs, whereas 50% of children were injured to upper limbs, face, and torso. Random wounds, typical of an explosion in vicinity, were observed in 32.9% of children and 18.7% of adults. All differences were statistically significant (P < 0.005). Time of evacuation, number of operations, and hospitalization did not statistically differ among groups. Number of transfused patients (23% vs. 7.2%), mortality (6.3% vs. 1.5%), incidence of blindness (21.2% vs. 9.5%), and maimed upper limbs (23.3% vs. 8.8%) were significantly higher in children compared with adults (P < 0.05). CONCLUSIONS: Long after ceasefire, antitank mines, antipersonnel land mines, and UXO continue to injure and kill civilians. Children are commonly injured and sustain more severe injuries. PMID- 19288283 TI - Preventing parastomal herniation: is prophylactic prosthetic mesh absolutely necessary? PMID- 19288284 TI - Clinical improvement after duodenojejunal bypass for nonobese type 2 diabetes despite minimal improvement in glycemic homeostasis. AB - BACKGROUND: Glycemic control of type 2 diabetes mellitus (T2DM) remains a dilemma to physicians. Although gastric bypass surgery undertaken for morbid obesity has been shown to resolve this disease well, data on the effectiveness of duodenojejunal bypass in improving or resolving T2DM and the metabolic syndrome (MS), especially in nonobese patients are scarce. This study was intended to evaluate the clinical effects of laparoscopic duodenojejunal bypass (LDJB) in patients with T2DM and a body mass index of <35 kg/m(2). METHODS: We conducted a 12-month prospective study on the changes in glucose homeostasis and the MS in seven T2DM subjects undergoing LDJB with similar DM duration, type of DM treatment, and glycemic control. Laboratory values including glycosylated hemoglobin A (HbA1c), fasting blood glucose, cholesterol, triglyceride, and C peptide were followed throughout the 12 months. Serum levels of gastric inhibitory peptide and ghrelin were followed for 1 month. Serum levels of gastrin and glucagon-like peptide were followed for 3 months. RESULTS: At 12 months after surgery, all subjects consistently felt relief from fatigue, pain and/or numbness in the extremities, polyuria, and polydypsia. Clinical resolution was obtained for one patient, and the preoperative diabetic medication requirements decreased for most of the other patients. The subjects demonstrated an overall improved HbA1c (from 9.4% to 8.5%) and fasting blood glucose level (from 209 to 154 mg/dl). Although the change in fasting blood glucose approached statistical significance, these measures of glucose homeostasis did not achieve significance. Cholesterol and triglycerides increased slightly, and C-peptide decreased slightly over 1 year. These changes were not statistically significant. CONCLUSIONS: Although this is a small series, our data show that at 12 months after surgery, clinical improvement was obvious in all of our seven patients, but LDJB may not be effective at inducing remission of T2DM and the MS in certain patients undergoing this operation. This suggests that larger patient studies should be conducted, before concluding that surgery may offer clinical and biochemical resolution to a disease once treated only medically. Longer follow-up is required for better evaluation. PMID- 19288285 TI - Can hyperbaric oxygenation decrease doxorubicin hepatotoxicity and improve regeneration in the injured liver? AB - BACKGROUND/PURPOSE: Portal vein embolization is used in the treatment of hepatocellular cancer, with the purpose of enhancing resectability. However, regeneration is restricted due to hepatocellular injury following chemotherapeutics (e.g. doxorubicin). The aim of this study was to investigate whether hyperbaric oxygenation (HBO) can alleviate the hepatotoxicity of chemotherapy and improve regeneration in the injured liver. METHODS: Rats were allocated to four experimental groups. Group I rats were subjected to right portal vein ligation (RPVL); rats in groups II and III were administered doxorubicin prior to RPVL, with group III rats being additionally exposed to HBO sessions postoperatively; group IV rats was sham-operated. All rats were sacrificed on postoperative day 7, and liver injury was assessed by measuring alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Protein synthetic ability was determined based albumin levels and liver regeneration by the mitotic index (MI). RESULTS: The AST and ALT values of group II rats were significantly higher than those of group I, but not those of group III. Rats treated with doxorubicin and HBO (groups II and III) showed slightly but not significant differences in albumin levels than those subjected to only RPVL or sham-operated. The MI was significantly increased in groups I, II, and III, with the MI of group III rats significantly higher than those of group I rats. CONCLUSIONS: Based on our results, we conclude that HBO treatment has the potential to diminish doxorubicin-related hepatotoxicity and improve regeneration in the injured liver. PMID- 19288286 TI - Renal allograft tuberculosis: report of three cases and review of literature. AB - Renal transplant recipients are prone to a variety of infections due a persistent immunodepleted state. Incidence of tuberculosis in this population is much higher compared with the general population. While pulmonary tuberculosis still remains the commonest form in this population, renal allograft tuberculosis is very rare. We report two cases of isolated allograft tuberculosis and one case of allograft tuberculosis with coexistent pleuro-pulmonary and bone marrow involvement. All three cases had presented with pyrexia of unknown origin, wherein despite extensive investigations the cause was not found. In two cases the diagnosis was confirmed on histology. Two cases responded to non-rifampicin-based modified antitubercular treatment and one to conventional four-drug Rifampicin-based regimen. Graft function improved in two cases while in one case the graft was lost. Tuberculosis involving the renal allograft is a potential cause for graft dysfunction/loss and requires a high index of suspicion for diagnosis. Timely detection and early institution of therapy can help save the renal allograft. PMID- 19288287 TI - Domestic violence: variation in case-management by the general practitioner in Belgium. AB - OBJECTIVES: to describe the medical interventions and referrals carried out by the general practitioner (GP) when taking care of victims and to quantify the between-physician variability in management of domestic violence. METHODS: A nationwide sentinel network of 150 general practitioners, covering 1.5 % of the Belgian population, registered in 2002-2004 all episodes of domestic violence for which they were consulted, via paper registration forms. A multilevel analysis was carried out by fitting a random effects logistic regression model for every intervention/referral. RESULTS: The most frequent interventions of the GP were providing a certificate of injury (54 %), and making an appointment for a next visit (33 %). Half of the patients were referred or hospitalised upon the first consultation, most frequently they were advised to go to the police (17 %) or referred to a psychologist or psychiatrist (11 %). The intra cluster correlation coefficient (ICC) of the interventions varied between 11 % and 39 % and the median odds ratios between 1,82 and 3,96. CONCLUSIONS: GP consultations for domestic violence are frequent and involve considerable between-physician variability in care. PMID- 19288288 TI - Can research contribute to the public's capacity for activities that reduce socioeconomic inequalities in health? PMID- 19288289 TI - From social determinants to reducing health inequalities. PMID- 19288290 TI - Seafaring stressors aboard merchant and passenger ships. AB - OBJECTIVES: The aim of this study was to identify stressors in seafaring aboard merchant and passenger vessels. Furthermore, their dependence on occupational and non-occupational factors was assessed. METHODS: A total of 134 male seafarers sailing under German-flagged vessels were interviewed (response 81.3 %). The seamen rated the individual stress level of 23 different stressors aboard. RESULTS: Separation from their family (named 48 times), time pressure (30 times), long working days (28 times), heat in workplaces (24 times), and insufficient qualification of subordinate crew members (16 times) were regarded as the most important stressors aboard. In comparison to non-officers, officers stayed on board for considerably shorter periods (4.8 vs. 8.3 months) but had significantly more often an extremely high number of working hours (63.5 % vs. 21.1 %, Chi square-test: p < 0.001). Correspondingly, officers complained more frequently of a higher stress level due to time pressure (52.4 % vs. 36.6 %). CONCLUSIONS: Particular attention should be paid to preventive organizational measures such as avoiding long-time separation from family, time-pressure, extremely long working days, and a long stay on board. PMID- 19288291 TI - Ongoing DNA synthesis in the rat cerebral cortex is regulated by a proteolytic pathway independent of the proteasome and calpains. AB - By using mini-units of tissue and protease inhibitors in short term incubation (0 180 min), we studied the role of proteolysis for ongoing DNA replication in the developing rat cerebral cortex. The protease inhibitors TLCK, TPCK, PMSF, MG-132 and PSI markedly inhibited DNA synthesis. The inhibitory effects were concentration-dependent and of early onset (within 60 min). The most selective proteasome inhibitors lactacystin and clasto-lactacystin-beta-lactone as well as the calpain inhibitor I and II had no or minimal effects on DNA synthesis. Only high concentrations of calpain inhibitor I (>or= 250 microM) and calpain inhibitor II (>or= 500 microM) gave a DNA synthesis inhibition. These results suggest that (1) ongoing DNA replication is regulated by proteolysis and (2) the proteolytic pathways involved are neither the proteasome nor the calpains. PMID- 19288292 TI - Molecular characterization of rice OsBIANK1, encoding a plasma membrane-anchored ankyrin repeat protein, and its inducible expression in defense responses. AB - A rice gene, OsBIANK1, encoding a protein containing a typical ankyrin repeat domain, was cloned and identified. The OsBIANK1 protein, consisting of 329 amino acids, contains a conserved ankyrin repeat domain with two ankyrin repeats organized in tandem and was showed to be localized on cytoplasmic membrane during transient expression in onion epidermal cells. Expression of OsBIANK1 was induced by treatment with benzothiadiazole (BTH), a chemical inducer capable of inducing disease resistance response in rice. In BTH-treated rice seedlings, expression of OsBIANK1 was further induced by infection with Magnaporthe grisea, the rice blast fungus, as compared with those in water-treated seedlings. Our preliminary results confirm previous evidences that OsBIANK1 may be involved in regulation of disease resistance response in rice. PMID- 19288293 TI - Bronchoprovocation testing. AB - Airway smooth muscles of asthmatics tend to be hyperresponsive when provoked. The exaggerated bronchial constriction can be measured by the airflow limitation seen following bronchial provocation. Measuring bronchial hyperresponsiveness by broncho provocation testing is helpful in diagnosing and optimizing therapy. While numerous agents have been used to provoke a measurable airflow limitation, standardized protocols are available for only a few. This article aims to discuss the various methods that have been reported for bronchoprovocation testing. PMID- 19288294 TI - Conversion of intestinal bypass to Roux-en-Y gastric bypass: a case report and brief review. AB - Intestinal bypass procedures for the treatment of morbid obesity have been largely replaced by Roux-en-Y gastric bypass procedures. The main reason for this change over the past 40 years stems from the myriad of negative nutritional and physiologic repercussions of intestinal bypass procedures. We present a case of a patient with severely atrophied small bowel and the novel method used for the conversion of the intestinal bypass procedure to Roux-en-Y gastric bypass. Additionally, we present a brief review of jejunoileal bypass. PMID- 19288295 TI - Bioavailability and health effects of cocoa polyphenols. PMID- 19288296 TI - Oppositional defiant and conduct disorder behaviors in boys with autism spectrum disorder with and without attention-deficit hyperactivity disorder versus several comparison samples. AB - We compared disruptive behaviors in boys with either autism spectrum disorder (ASD) plus ADHD (n = 74), chronic multiple tic disorder plus ADHD (n = 47), ADHD Only (n = 59), or ASD Only (n = 107). Children were evaluated with parent and teacher versions of the Child Symptom Inventory-4 including parent- (n = 168) and teacher-rated (n = 173) community controls. Parents rated children in the three ADHD groups comparably for each symptom of oppositional defiant disorder (ODD) and conduct disorder. Teacher ratings indicated that the ASD + ADHD group evidenced a unique pattern of ODD symptom severity, differentiating them from the other ADHD groups, and from the ASD Only group. The clinical features of ASD appear to influence co-morbid, DSM-IV-defined ODD, with implications for nosology. PMID- 19288298 TI - Modification of biodegradable poly(malate) and poly(lactic-co-glycolic acid) microparticles with low molecular polyethylene glycol. AB - Poly(lactic-co-glycolic acid) microparticles (PLGA-MP) and cross-linked poly(malate) microparticles (PMAL-MP) were prepared by solvent/extraction and polycondensation, respectively. Microparticles were modified with polyethylene glycol (PEG 400) which lowered the drug loading of PMAL-MP, but did not influence the loading of PLGA-MP. The drug release from PMAL-PEG-MP occurred by initial degradation to oligomeric fractions, followed by further hydrolysis of oligomers to free drug (dipyridamole). The complete release was achieved for 22 days, whereas the release from non-modified PMAL-MP took 38 days. The drug release from PLGA-PEG-MP was characterized with an initial burst effect, followed by slower release during 26 days. Thus, the modification with PEG could be used to achieve appropriate release of the selected antithrombotic drug. PMID- 19288297 TI - Increased accumulation of intraneuronal amyloid beta in HIV-infected patients. AB - In recent years, human immunodeficiency virus (HIV)-infected patients under highly active anti-retroviral therapy (HAART) regimens have shown a markedly improved general clinical status; however, the prevalence of mild cognitive disorders has increased. We propose that increased longevity with HIV-mediated chronic inflammation combined with the secondary effects of HAART may increase the risk of early brain aging as shown by intraneuronal accumulation of abnormal protein aggregates like amyloid beta (Abeta), which might participate in worsening the neurodegenerative process and cognitive impairment in older patients with HIV. For this purpose, levels and distribution of Abeta immunoreactivity were analyzed in the frontal cortex of 43 patients with HIV (ages 38-60) and HIV- age-matched controls. Subcellular localization of the Abeta immunoreactive material was analyzed by double labeling and confocal microscopy and by immunono-electron microscopy (EM). Compared to HIV- cases, in HIV+ cases, there was abundant intracellular Abeta immunostaining in pyramidal neurons and along axonal tracts. Cases with HIV encephalitis (HIVE) had higher levels of intraneuronal Abeta immunoreactivity compared to HIV+ cases with no HIVE. Moreover, levels of intracellular Abeta correlated with age in the group with HIVE. Double-labeling analysis showed that the Abeta-immunoreactive granules in the neurons co-localized with lysosomal markers such as cathepsin-D and LC3. Ultrastructural analysis by immuno-EM has confirmed that in these cases, intracellular Abeta was often found in structures displaying morphology similar to autophagosomes. These findings suggest that long-term survival with HIV might interfere with clearance of proteins such as Abeta and worsen neuronal damage and cognitive impairment in this population. PMID- 19288299 TI - An assessment of metal recycling worker lead exposure associated with cutting uncoated new steel scrap. PMID- 19288301 TI - Ratiometric Ca+2 measurement in human recombinant muscarinic receptor subtypes using the Flexstation scanning fluorometer. AB - In modern drug discovery, numerous assay formats are available to screen and quantitate receptor-ligand interactions. Radioactive assays are "gold standard" because they are fast, easy, and reproducible; however, they are hazardous, produce radioactive waste, require special lab conditions, and are expensive on a large scale. Thus, it provides a lot of importance to the "mix & measure" assays that have an optical readout. Fluorescence techniques are likely to be among the most important detection approaches used for high throughput screening due to their high sensitivity and amenability to automation. The aim of the present study was to determine the functional antagonistic affinities of standard muscarinic antagonists in CHO cells over expressing m1, m3, and m5 receptors and to compare them with the respective binding affinities. This study was further extended to elucidate that Ca+2 measurement assays can serve as a functional screening tool for GPCRs. For this purpose, standard muscarinic receptor antagonists, namely, tolterodine, oxybutynin, and atropine were used. We determined and compared the IC50 values of these three standard inhibitors in fura 2 AM loaded m1, m3, and m5 overexpressing CHO cells and in radioligand binding assay. Both the assays exhibited comparable rank order potencies of the standard inhibitors. This study suggests that Ca+2 mobilization assays can be an alternate to radioligand binding assays. PMID- 19288302 TI - A theory of the origin of cerebral asymmetry: epigenetic variation superimposed on a fixed right-shift. AB - A theory of the genetic basis of cerebral asymmetry is outlined according to which (1) a single right-shift factor in all human individuals interacts with (2) epigenetic variation that is apparently random, transmissible between parent and child, but with a half-life extending over a small number of generations. The right-shift factor arose late (150 to 200 thousand years ago [KYA]) in hominid evolution as a mutation in the X copy of a gene pair (Protocadherin 11XY) in a region of homology between the X and Y chromosomes created by a duplication 6MYA at the chimpanzee hominid separation. The epigenetic imprint originates in the process now known as "meiotic suppression of unpaired chromosomes" (MSUC); it reflects random pairing of partly homologous X and Y chromosome regions in male meiosis, and confers species-specific diversity of cerebral structure on the human population. PMID- 19288305 TI - Virtual human dissector as a learning tool for studying cross-sectional anatomy. AB - BACKGROUND: Within diagnostic medicine there is a continuing and marked increase in the use of two-dimensional (2D) images of cross-sectional anatomy. Medical undergraduates should therefore develop skills to interpret such images early in their education. The Virtual Human Dissector (VHD) software facilitates such learning, permitting users to study actual images of 2D anatomical cross-sections and reconstructed three-dimensional (3D) views simultaneously. This study investigates the use of VHD in facilitating students' ability to interpret cross sectional images and understand the relationships between anatomical structures. METHODS: First year medical students (n=89) were randomly divided into two groups. Using a crossover design, the investigation was undertaken as two 20 minute self-directed learning (SDL) activities using VHD in a computer suite and prosections and models in the dissecting room (DR), interspersed between 3 tests identifying anatomical structures in cross-sectional images (pre-, mid- and post session). RESULTS: Statistical analysis of test performance revealed significant improvements in each group between the pre- and midsession tests, and again between mid- and post-session tests. There was no significant difference between the two groups at any stage. SDL using the VHD was as effective as SDL using prosections. PMID- 19288306 TI - The Ventriloscope: 'am I hearing things?'. AB - BACKGROUND: Knowledge of the benefits of incorporating medical simulation into healthcare curricula is rapidly increasing. Though impeded by the high cost of complicated technology, medical simulation devices offer the ability to provide safe and controlled training environments, exposure to rare clinical scenarios, as well as unlimited training opportunities. METHODS: This report describes a novel, inexpensive method of broadcasting normal and abnormal auscultatory findings to a relatively normal appearing stethoscope for use in training of healthcare professionals. RESULTS: Using wireless transmitter broadcasting to a stethoscope fitted with a receiver apparatus, the student is able to perform a typical medical exam with auscultation of an unlimited variety of clinical sounds from anatomically appropriate sources while being observed from another room. CONCLUSIONS: Implications of this low-cost device include limitless training possibilities worldwide and across disciplines. The simplicity and portability of this device increases potential for use in rapid training of recognition of clinical signs associated with chemical/biological warfare agents, mass casualty incidents and field military applications. This is the first device to simulate clinically relevant sounds in a realistic manner on standardized patients and mannequins. The benefits of such simulation in medical education ultimately serve to increase trainee confidence and consequently, improve patient care and safety. PMID- 19288307 TI - Tetrahedron of medical academics: reasons for training in management, leadership and informatics. AB - Medical school professors and lecturers are often called to be practicing clinicians, researchers in their own field, in addition to executing their education and curricular responsibilities. Some further accumulate healthcare management responsibilities. These areas pose conflicting demands on time and intellectual activity, but despite their apparent differences, knowledge and skills from management, leadership and informatics may prove useful in helping to smooth these conflicts and hence increase personal effectiveness in these areas. This article tries to clarify some concepts and advance why training in management, leadership and health informatics would seem particularly useful for the medical academic. As opposed to the idea of educational dispersion/specialization, the concept of an integrative tetrahedronal education framework is advanced as a way to plan workshops and other faculty development activities which could be implemented transnationally as well as locally. PMID- 19288308 TI - Placing learning needs in context: distance learning for clinical officers in Tanzania. AB - BACKGROUND: Poor public health indicators in Tanzania have led to the upgrading of nursing and clinical personnel who currently have just core training. Clinical officers (COs) have 3 years training in basic and applied medicine and are responsible for healthcare of large and dispersed rural populations. AIMS: UNESCO Wales has funded colleagues in Wales (UK) to assist the upgrade of COs. An inquiry into their learning needs and the Tanzanian context has produced a framework for design of a module for COs on sexually transmissible infections and HIV & AIDS by distance learning. METHODS: Face-to-face discussions were held with the Ministry of Health, healthcare workers, educators and administrators in Tanzania; a review of training documents was carried out; and a follow-up questionnaire issued to COs. RESULTS: The discussions and review highlighted teacher-centred approaches, and management, infrastructure and resources obstacles to curriculum change. Principal learning needs of COs around STIs were: counselling, syndromic management, drugs management, laboratory diagnosis, health education, resources, staffing and service morale. CONCLUSIONS: Placing learning needs in context in dialogue with Tanzanian colleagues was an advance on simple transfer of educational technologies and expertise. The inquiry resulted in a draft study guide and resources pack that were positively reviewed by Tanzanian tutors. Management and resources issues raised problems of sustainability in the module implementation. PMID- 19288309 TI - Variations in medical school graduating examinations in the United Kingdom: are clinical competence standards comparable? AB - BACKGROUND: While all graduates from medical schools in the UK are granted the same licence to practise by the medical professional regulatory body, the General Medical Council, individuals institution set their own graduating examination systems. Previous studies have suggested that the equivalence of passing standards across different medical schools cannot be guaranteed. AIMS: To explore and formally document the graduating examinations being used in the UK Medical Schools and to evaluate whether it is possible to make plausible comparisons in relation to the standard of clinical competence of graduates. METHODS: A questionnaire survey of all the UK medical schools was conducted, asking for details of graduating examination systems, including the format and content of tests, testing time and standard setting procedures. RESULTS: Graduating assessment systems vary widely across institutions in the UK, in terms of format, length, content and standard setting procedures. CONCLUSIONS: We question whether is it possible to make plausible comparisons in relation to the equivalence of standards of graduates from the different UK medical schools, as current quality assurance systems do not allow for formal quantitative comparisons of the clinical competence of graduates from different schools. We suggest that national qualifying level examinations should be considered in the UK. PMID- 19288311 TI - Can we create an equivalent educational experience on a two campus medical school? AB - BACKGROUND: Does the university experience and environment on two very different campuses create perceptions of advantage or disadvantage despite delivery of the same curriculum by the same tutors? Perhaps more importantly, do different types of universities result in varying achievements in learning given similar students? AIMS: The Hull York Medical School (HYMS) runs the same curriculum with the same faculty on two very different campuses and randomly allocates most students to one or the other. HYMS therefore offers an exceptional opportunity to investigate these questions about the perceptions of educational environment and actual academic achievement controlling as much as possible for design and delivery of the curriculum. METHOD: The Dundee Ready Education Environment Measure (DREEM) was administered to students in Year 1 and 2 at HYMS to assess perceptions of the course. Examination results were collected for the cohorts to compare actual academic performance on written and clinical examinations. RESULTS: Minimal differences were found in perceptions of educational environment, with those differences found favouring the less 'prestigious' institution. Examination results for written and clinical exams over the first 2 years were found not to differ between campuses. CONCLUSIONS: The results of this study indicate that despite perceptions of one university being 'better' than another based on public measures such as league tables, the students' perceptions of educational environment was quite similar and exam performance showed no differences. This suggests that the prestige or ranking of a university based on common measures and perceptions may have far less impact on student learning than careful design and delivery of the curriculum. PMID- 19288312 TI - Change in senior medical students' attitudes towards the use of mathematical modelling as a means to improve research skills. PMID- 19288314 TI - A Faculty Development Program can result in an improvement of the quality and output in medical education, basic sciences and clinical research and patient care. AB - The Carl Gustav Carus Faculty of Medicine, University of Technology Dresden, Germany, was founded in 1993 after the reunification of Germany. In 1999, a reform process of medical education was started together with Harvard Medical International.The traditional teacher- and discipline-centred curriculum was displaced by a student-centred, interdisciplinary and integrative curriculum, which has been named Dresden Integrative Patient/Problem-Oriented Learning (DIPOL). The reform process was accompanied and supported by a parallel-ongoing Faculty Development Program. In 2004, a Quality Management Program in medical education was implemented, and in 2005 medical education received DIN EN ISO 9001:2000 certification. Quality Management Program and DIN EN ISO 9001:2000 certification were/are unique for the 34 medical schools in Germany.The students play a very important strategic role in all processes. They are members in all committees like the Faculty Board, the Board of Study Affairs (with equal representation) and the ongoing audits in the Quality Management Program. The Faculty Development program, including a reform in medical education, the establishment of the Quality Management program and the certification, resulted in an improvement of the quality and output of medical education and was accompanied in an improvement of the quality and output of basic sciences and clinical research and interdisciplinary patient care. PMID- 19288316 TI - Teaching in the clinical environment: Guide supplement 34.2-Viewpoint. PMID- 19288317 TI - Students' perceptions regarding educational environment in an Indian dental school. AB - BACKGROUND: As one of India's premier dental schools, the Manipal College of Dental Sciences, Manipal, has striven to be in the forefront of dental education ever since its inception in 1965. But one of our main drawbacks has been the lack of students' perceptions about the educational atmosphere in our institution. AIM: (1) To understand the students' perceptions of the educational environment. (2) To compare the results with those of other dental/medical schools. METHODS: Dundee Ready Education Environment Measure (DREEM) was used to gather information about the environment in our institution. The same was administered to first year and final year students (63 each). RESULTS: The main area of convergence of both groups was that the social atmosphere was not very congenial. But the thought provoking finding was that both groups felt that the teachers were authoritarian. CONCLUSION: The lack of congenial atmosphere in our institution seems to be due to the attitude of the course organizers.A need for makeover in attitudes and approach on the part of the teachers to make the learning experience more pleasant for the students was made obvious by this study. PMID- 19288319 TI - Improving rational prescribing by medical students. PMID- 19288318 TI - Smoking and exercise among future physicians: survey of knowledge, attitudes and behavior of students at a faculty of medicine in Israel. PMID- 19288321 TI - Comparison of iodixanol and iohexol in patients undergoing intravenous pyelography: a prospective controlled study. AB - BACKGROUND: Nephropathy associated with contrast medium exposure is a well-known complication of IVP. However, it is uncertain whether iso-osmolar non-iodinated contrast medium (iodixanol) is less nephrotoxic than low-osmolar contrast medium (iohexol). MATERIALS AND METHODS: In this single-center, double-blind, prospective study, 50 patients undergoing IVP were randomized into two groups receiving different contrast medium: iodixanol and iohexol. Patients in high risk for contrast nephropathy were included, 28 with renal insufficiency and 19 with diabetes mellitus. We compared the nephrotoxic effect (contrast nephropathy), complement and cytokines profile between the iodixanol and iohexol groups. The mean volume of contrast medium in each IVP procedure was 0.8 mL/kg. RESULTS: The incidence of contrast nephropathy was 4 percent among all patients (one iodixanol and one iohexol). We found no significant differences in contrast nephropathy and allergic reactions between the two groups. There was no significant difference in cytokine profiles in both groups (p > 0.05).The incidence of allergic reaction was 16 percent among all patients. Twelve percent (3/25) had late reaction after iohexol exposure compared to four percent (2/25) with iodixanol (p = 1.0). One patient had severe skin rash due to late adverse reaction after iodixanol. No mortality was found. CONCLUSIONS: New iodixanol and iohexol contrast medium for routine IVP examination are safe and have low nephrotoxicity profile, especially in elderly or high-risk patients. Iodixanol contrast medium has an increased risk to induce severe late adverse reaction compared to iohexol. Allergic reaction may be the main adverse effect after contrast medium infusion. PMID- 19288322 TI - Achieving KDOQI Guidelines for Hematocrit in HIV-Infected Hemodialysis Patients. AB - The primary cause of anemia in HIV-infected patients with ESRD is diminished production of erythropoietin. Although most patients respond to recombinant erythropoietin, the response may be blunted in patients with ESRD and concomitant viral or bacterial infections. Previous studies demonstrated a response to erythropoietin by HIV-infected ESRD patients, but hematocrit levels on average were only 27-29%. We were interested in determining if KDOQI guidelines could be met in these patients. Hematocrits and epogen doses of all HIV-positive patients who were undergoing hemodialysis at the Nassau University Medical Center Dialysis Unit between September 2002 and March 2003 were compared to matched controls in our hemodialysis unit. The hematocrit levels in our population were higher than those reported in earlier papers. In our patient population, the mean hematocrit was 37.5, whereas the mean hematocrit levels in the HIV group in previous papers were 27-29%. HIV-infected patients did require higher erythropoietin dosages than controls, but similar doses were used as compared to previous studies. HIV patients on hemodialysis can achieve KDOQI target hematocrits. The difference in route of iron administration and iron stores may explain the higher hematocrit levels in our HIV patient population as compared to previous trials. PMID- 19288323 TI - The role of pure diffuse mesangial hypercellularity in patients with proteinuria. AB - BACKGROUND: Pure diffuse mesangial hypercellularity (DMH), in its primary form, is a relatively rare histological finding, and scant data exist in the literature regarding its clinical course and prognosis in nephrotic adults with this diagnosis. METHODS: We retrospectively analyzed the clinical and histological data of 8 out of 41 patients with the above diagnosis in regard to response to the treatment, outcome and prognostic indicators. RESULTS: Six patients received oral prednisolone as initial therapy, five of whom receiving it as monotherapy at first. The two other patients did not receive anything at all. Three out of the above six patients received prednisolone either with cyclophosphamide or with cyclosporine (CyA). Three patients responded with complete remission, two showed partial remission, and one did not respond at all. During follow-up, none of the patients with complete response appeared to have relapse. The two patients with initial partial response to steroids received CyA in combination with low dose of oral prednisolone. The other patient who did not respond at all from the beginning did not receive anything more due to his bad general condition. Plasma creatinine remained stable in those with complete or partial response to treatment. None of the clinical characteristics was found to be predictive of the degree of renal function impairment at the time of renal biopsy. The three patients with partial or no response were characterized by the severity of mesangial hypercellularity. Patients with complete or partial response to therapy did not differ with regard to age, plasma creatinine, and severity of proteinuria at biopsy. Presence of mesangial IgM was not associated with poor or satisfactory response. In general, no clinical feature at the time of biopsy was predictive of a response to therapy. CONCLUSIONS: At present, it seems that adult patients with DMH and proteinuria represent a heterogeneous group with different clinical courses despite a similar morphological appearance in initial biopsies. We conclude that serial biopsies taken at regular intervals coupled with a longer follow-up may provide answers concerning the real intensity of DMH. PMID- 19288324 TI - Renin-angiotensin system polymorphisms: a risk factor for progression to end stage renal disease in vesicoureteral reflux patients. AB - Aim. Renin-angiotensin system (RAS) gene mutations have been implicated as a risk factor for the presence and progression of renal disease in vesicoureteral reflux (VUR). However, the results are contradictory, and the effects of RAS polymorphisms in VUR patients with end-stage renal disease (ESRD) have not been defined yet. This study was designed to evaluate the angiotensin-converting enzyme insertion/deletion (ACE-I/D), angiotensinogen (AGT) M235T, and angiotensin II receptor type 1 (ATR1) A1166C and type 2 (ATR2) C3123A gene polymorphisms as risk factors for progression to ESRD in patients with VUR. Methods. ACE-I/D, AGT M235T, ATR1-A1166C, and ATR2-C3123A were identified in 161 ESRD patients (52 female, 109 male; 77 renal transplant, 84 dialysis; age: 34.4 +/- 11.2 years). VUR was the ESRD etiology in 40 patients. Genetic polymorphisms of the ACE gene I/D, AGT gene M235T, ATR1 gene A1166C, and ATR2 gene C3123A were identified in all of the patients. Results. We detected no linkage between genetic polymorphisms of ATR1-, ATR2-, AGT-, and VUR-related ESRD. When ACE gene was considered, VUR(+) patients had 63.6% DD, 36.4% ID, and no II alleles, whereas VUR(-) patients had 48.6% DD, 43.2% ID, and 8.1% II alleles. Conclusion. A striking feature of VUR-related ESRD patients was the absence of II alleles, so the DD genotype may be accepted as a genetic susceptibility factor for progression to ESRD in VUR patients. PMID- 19288325 TI - Health-related quality of life, treatment efficacy, and hemodialysis patient outcome. AB - The aim of the study was to examine the influence of improved treatment of hemodialysis (HD) patients on their health-related quality of life (HrQoL) and to assess the predictive value of HrQoL dimensions on patient outcome. The prospective cohort study involved 102 HD patients, and their clinical and laboratory parameters and HD adequacy indices were followed from 2001 to 2007. HrQoL was measured using KDQOL-SF Version 1.3 in 2001, 2004, and 2007. During a six-year period, quality of HD and anemia treatment improved and resulted in significant increase of mean Kt/V (1.2-1.56) and hemoglobin levels (86.5-115.6 g/L). All four HrQoL dimensions (i.e., physical, mental health, kidney disease target issues, and patient satisfaction) remained unchanged, but significant improvement in several HrQoL physical health domains and the effects of kidney disease domain was found. Mortality rate decreased from 18.6% to 7.14% per year. Age was associated positively, but kidney disease target issue score negatively, with patient death. Improved HD adequacy and anemia treatment in HD patients were followed with maintenance of all four HrQoL dimensions unchanged over six years. Moreover, an improvement in several physical health domains and the effects of kidney disease domain was found. Age and kidney disease target issue appeared as significant predictors of patients' death. PMID- 19288326 TI - Altered vascular homeostasis in type 2 diabetic nephropathy. AB - Type 2 diabetic nephropathy is a primary cause of ESRD worldwide. Therapeutic strategy in patients with microalbuminuric or macroalbuminuric type 2 diabetic nephropathy usually fails to restore renal function but merely slows the renal disease progression. In contrast, a recent study implies that the restoration of renal function as well as renal perfusion can be accomplished in early stage of type 2 diabetic nephropathy (normoalbuminuria) by correcting the hemodynamic maladjustment in renal microcirculation with vasodilators. Therefore, we intend to study the mechanism of vascular homeostasis to explain why treatment in the late stage of diabetic nephropathy during microalbuminuria or macroalbuminuria fails to enhance renal perfusion or restore renal function. The results indicate that such therapeutic failure in late-stage type 2 diabetic nephropathy likely relates to multiple defects in vascular repair, namely deficiencies in angiogenic factors such as endothelial progenitor cell, angiopoietin-1, flt-1 receptor, as well as elevated levels of antiangiogenic factors such as angiopoietin-2 and KDR. PMID- 19288327 TI - Cardiac function in an African dialysis population with a low prevalence of pre existing cardiovascular disease. AB - Cardiac dysfunction is highly prevalent in dialysis patients in the developed world, and is a major cause of morbidity and mortality. The relative impact of pre-existing cardiac disease and dialysis/uremia on cardiovascular morbidity are not clear. We conducted a retrospective, cross-sectional analysis of cardiac function and mortality in 202 incident and prevalent dialysis patients over an 18 month period in a population with a low prevalence of cardiovascular disease at dialysis initiation. Systolic dysfunction was defined as an ejection fraction (EF) of <50%. Left ventricular hypertrophy (LVH) was determined by echocardiography or electrocardiogram. Clinical data was collected by chart review. Ninety-nine percent of patients were black, with a mean age of 41.7 +/- 10.1 years, and median follow up 28 months (range 1-216 months). Echocardiograms were available in 132 patients. Seventy-seven patients received hemodialysis, and 55 received peritoneal dialysis. Mean EF was 63.2 +/- 11.1. EF was not lower in patients with greater duration of dialysis, although LVH tended to increase (not statistically significant). In 39 patients who died during the study period, cardiac function was not different from survivors, and no patient died of ischemic heart disease or heart failure. In conclusion, in a population of patients with a low prevalence of cardiovascular disease at dialysis initiation, cardiac function appears preserved over time, and cardiac morbidity and mortality are low. This finding suggests that dialysis and uremia per se, in the absence of pre-existing cardiac disease, may not be major contributors to cardiovascular morbidity. PMID- 19288328 TI - Frequency of metabolic syndrome among hemodialysis patients according to NCEP-ATP III and IDF definitions. AB - Objective. Patients with chronic renal failure (CRF) have an increased risk of death from cardiovascular diseases. The metabolic syndrome is a common risk factor for cardiovascular diseases. In the present study, it was aimed to evaluate the frequency of metabolic syndrome using the National Cholesterol Education Program Adults Treatment Panel III (NCEP-ATP III) and the International Diabetes Federation (IDF) definitions in patients with end-stage CRF undergoing hemodialysis (HD). Materials and Methods. A total of 222 cases undergoing HD were enrolled in the study. After obtaining medical history and physical examination, blood samples were collected from each patient for the measurements of fasting blood glucose, total cholesterol, high-density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol, and triglycerides. Results. Among HD patients evaluated according to both IDF and NCEP-ATP III definitions, the diagnosis of metabolic syndrome was confirmed by IDF in 56.5% of those fulfilling the criteria for NCEP-ATP III. Similarly, 86% of the undiagnosed patients according to NCEP-ATP III were confirmed by IDF definitions. The sensitivity and positive predictive value of NCEP-ATP III for metabolic syndrome were 81.25% and 64.8%, respectively. The area under the Receiver Operating Characteristic (ROC) curve for NECP-ATP III and IDF was 0.730. False-positive rate and probability ratio for NECP-ATP III were 0.352 and 2.49, respectively. In other words, among the patients who were diagnosed with metabolic syndrome according to NCEP-ATP III definitions, the proportion of subjects whose diagnosis was confirmed by IDF definitions was 2.49-fold higher than those with unconfirmed diagnosis. Conclusion. It is logical to evaluate patients with CRF for metabolic syndrome and cardiovascular risk factors at the time of diagnosis and regularly thereafter due to the high ratio of metabolic syndrome in this population. PMID- 19288329 TI - Proteinuria-induced chronic kidney disease in the ICGN/Oa mice with a mutation of Tensin2 gene. AB - ICGN/Oa mice are used to study the pathophysiological mechanisms underlying proteinuria-induced chronic kidney disease (CKD). Recently, a mutation of tensin2 gene (Tns2) was suggested to be responsible for proteinuria in the inbred ICGN mice. We identified the wild-type (+/+), heterozygous (+/nep), and homozygous (nep/nep) ICGN/Oa mice by PCR assay. The homozygotes developed proteinuria, resulting in nephrotic syndrome (NS) as early as 5 weeks and CKD by 15 weeks. However, the heterozygotes did not show the symptoms of these renal failures. These results indicate that the homozygous tensin2 mutation is necessary for the ICGN/Oa mice to develop proteinuria-induced CKD. Furthermore, we examined the time course of tubulointerstitial fibrosis and the kinetics of tubular epithelial cells (TECs) in the ICGN/Oa mice using immunohistochemical and TUNEL assays. In the renal parenchyma of the five-week-old homozygotes, the expression of alpha SMA and type I collagen were higher than those in the age-matched wild-type. Additionally, increased TEC proliferation was found at 5 weeks, and increased TEC apoptosis was by 15 weeks in the homozygotes. Tubulointerstitial fibrosis precedes TEC apoptosis in the proteinuria-induced CKD model mice, and that tubulointerstitial fibrosis may be the triggering event of the disease. PMID- 19288330 TI - Malnutrition in the elderly patient on dialysis. AB - Approximately one-third of all dialysis patients have mild to moderate malnutrition, while 6-8% have severe malnutrition, which is associated with increased morbidity and mortality rates and numerous pre-existing factors directly correlated with, or existing prior to, replacement hemodialysis. However, moderate to severe malnutrition (present in 10-30% of dialysis patients) is a prevalent cause of death among the elderly. Many of these patients have a particularly unstable cardiovascular and metabolic status that, independent of any underlying uremia and/or dialysis, impacts negatively on both their quality of life and clinical status. Moreover, their condition is often further exacerbated by dialysis itself, with its acute (e.g., hypotension and sensorial alterations) and chronic complications, including an exacerbation of malnutrition and systemic vascular disease. Malnutrition can occur secondary not only to erroneous dietary choices or uremia, but it may also depend on the patient's level of tolerance to dialysis and on the dialysis modality. Despite the improvements made to dialysis techniques, the nutritional condition of elderly patients on dialysis for chronic renal failure remains a cause for concern. In this patient category, it is therefore mandatory to ensure the daily supervision of nutritional status and early control when the first signs of malnutrition appear. PMID- 19288331 TI - Disseminated lymphoma presenting as acute thigh pain and renal failure. AB - A 66-year-old diabetic man presented with severe right thigh swelling and pain together with acute renal failure. At autopsy, this was found to be due to disseminated high grade B cell lymphoma invading the psoas muscle and multiple organs, including the kidneys. The unique presentation of this case emphasizes the need for increased awareness of the variety of ways in which lymphoma can manifest itself. PMID- 19288332 TI - Esophageal tuberculosis in a patient on maintenance dialysis: advantages of interferon-gamma release assay. AB - Patients with chronic renal failure have a higher risk of developing tuberculosis compared to the general population, and especially extra-pulmonary tuberculosis. This pathology is often difficult to diagnose and requires a combination of multidisciplinary examinations. The confirmation of the diagnosis is important in order to quickly match the treatment with the pathology. We report a patient with primary esophageal tuberculosis for whom the interferon gamma release assay facilitated a timely diagnosis. PMID- 19288333 TI - Comparison of assessment measures for post-stroke depression. AB - Depression is commonplace following stroke and is associated with increased morbidity and mortality. The objective of this study is to examine the sensitivity and specificity of four standardized measures for assessing depression in a stroke population. A total of 67 adults admitted to an inpatient rehabilitation unit with acute stroke were assessed with four standardized depression measures and by structured clinical interview as part of a cross sectional study assessing cognitive and emotional functioning. Using DSM-IV criteria, major depression was present in 15% of the sample and minor depression in an additional 28%. The four depression measures yielded rates of depression ranging from 14% to 46% when their recommended clinical threshold scores were applied. The Center for Epidemiologic Studies Depression Scale (CES-D) had the greatest positive predictive value (PPV) at 28%. Cutoff scores were adjusted using receiver operating curve (ROC) analyses, and PPV ranged from 28% to 34% when using lower cutoff scores, representing an improvement for all measures. Common assessment measures yield significantly different classification rates for depression in persons with acute stroke. Traditional threshold scores for commonly used objective depression measures may not be optimally sensitive for detection of depression in stroke populations. Based on the findings of this study we recommend clinicians use the Geriatric Depression Scale-Short Form using a cutoff score of 3 or greater. Clinical and research implications are offered. PMID- 19288334 TI - Response times in organic vapor instruments. PMID- 19288335 TI - Validity and reliability of an occupational exposure questionnaire for parkinsonism in welders. AB - This study assessed the validity and test-retest reliability of a medical and occupational history questionnaire for workers performing welding in the shipyard industry. This self-report questionnaire was developed for an epidemiologic study of the risk of parkinsonism in welders. Validity participants recruited from three similar shipyards were asked to give consent for access to personnel files and complete the questionnaire. Responses on the questionnaire were compared with information extracted from personnel records. Reliability participants were recruited from the same shipyards and were asked to complete the questionnaire at two different times approximately 4 weeks apart. Percent agreement, kappa, intraclass correlation coefficient (ICC), and sensitivity and specificity were used as measures of validity and/or reliability. Personnel files were obtained for 101 of 143 participants (70%) in the validity study, and 56 of the 95 (58.9%) participants in the reliability study completed the retest of the questionnaire. Validity scores for items extracted from personnel files were high. Percent agreement for employment dates and job titles ranged from 83-100%, while ICC for start and stop dates ranged from 0.93-0.99. Sensitivity and specificity for current job title ranged from 0.5-1.0. Reliability scores for demographic, medical and health behavior items were mainly moderate or high, but ranged from 0.19 to 1.0. Most recent job/title items such as title, types of welding performed, and material used showed substantial to perfect agreement. Certain determinants of exposure such as days and hours per week exposed to welding fumes demonstrated mainly moderate agreement (kappa= 0.42-0.47, percent agreement 63 77%); however, mean days and hours reported did not differ between test and retest. The results of this study suggest that participants' self-report for job title and dates employed are valid compared with employer records. While kappa scores were low for some medical conditions and for caffeine consumption, high kappa scores for job title, dates worked, types of welding, and materials welded suggest participants generated reproducible answers important for occupational exposure assessment. PMID- 19288336 TI - Sexual behaviors and attitudes and ethnic identity during college. AB - This study examined ethnic differences in sexual behaviors and attitudes, and associations between ethnic identity commitment and sexual behaviors and attitudes. African American (32%), Latino American (29%), and European American (39%) first-year college students (N = 434; 52% female) completed surveys about their sexual behaviors (number of partners, condom use, and alcohol use before intercourse) and attitudes (conservative attitudes, condom-related beliefs, and fear of AIDS) and ethnic identity commitment. Analyses of covariance and hierarchical linear regressions were performed. Among the three groups, Latino Americans reported riskier condom-related behaviors and attitudes, whereas European Americans were less fearful of AIDS. Ethnic identity commitment was a protective factor against risky attitudes regardless of ethnicity. For sexual behaviors, however, ethnic identity served as a protective factor only for European Americans. This study contributes to the understanding of adolescents' sexuality during college and the role of ethnic identity in their sexual experiences. PMID- 19288337 TI - Viva viva? Women's meanings associated with male nudity in a 1970s "for women" magazine. AB - Little research has been conducted with regard to how women view male nudity. The purpose of this analysis was to present an historical case study of Viva, a 1970s women's magazine geared toward the presentation of male nudity. Implicit in Viva's editorial direction was the assumption that women's sexuality is socially constructed and, thus, modifiable but also homologous to men's sexuality. Using sexual scripting theory as a sensitizing concept, a content analysis of women's letters to the editor was conducted. Letters justified inclusion of male nudity on the basis of principles of fairness and equality with men. In addition, letters tried to demarcate boundary conditions for what constituted acceptable male nudity in terms of the presence of erection, size, shape, and coloring of the penis, and whether inclusion of the penis was contextualized by other physical traits such as body musculature. Despite an early effort to modify cultural scripts on the basis of a constructivist world view, it was found that, ultimately, Viva advocated an essentialist viewpoint that reified existing conceptualizations of female sexuality as both static and uninterested in the visual representation of sexually related material. This study explored whether Viva's essentialist final position may have reflected financial exigencies rather than ideologies. PMID- 19288338 TI - Questionnaire of cognitive schema activation in sexual context: a measure to assess cognitive schemas activated in unsuccessful sexual situations. AB - The aim of this study was to assess the reliability and validity of the Questionnaire of Cognitive Schema Activation in Sexual Context (QCSASC; Nobre & Pinto-Gouveia, 2000b). A total of 319 individuals (96 with sexual dysfunctions and 223 without sexual dysfunctions) participated in the study. The QCSASC assesses 28 self-schemas proposed by J. S. Beck (1995), usually associated with psychological problems. The measure assesses the activation of these self-schemas following the presentation of 4 negative sexual events associated with the most common sexual dysfunctions in men and women. The QCSASC demonstrated internal consistency (Cronbach's alpha = .94), test-retest reliability (r[34] = .66), convergent validity (high correlations with other measures of general schemas), and incremental validity. Discriminant analysis differentiated between clinical (sexual dysfunction) and non-clinical groups, giving credit to the hypothesis that the activation of negative schemas may be involved in sexual dysfunctional processes. Findings also suggest the relevance of assessing cognitive schemas in clinical settings. PMID- 19288339 TI - Gender inequity in health: why it exists and how we can change it. PMID- 19288340 TI - The intersections of gender and class in health status and health care. AB - It is increasingly recognized that different axes of social power relations, such as gender and class, are interrelated, not as additive but as intersecting processes. This paper has reviewed existing research on the intersections between gender and class, and their impacts on health status and access to health care. The review suggests that intersecting stratification processes can significantly alter the impacts of any one dimension of inequality taken by itself. Studies confirm that socio-economic status measures cannot fully account for gender inequalities in health. A number of studies show that both gender and class affect the way in which risk factors are translated into health outcomes, but their intersections can be complex. Other studies indicate that responses to unaffordable health care often vary by the gender and class location of sick individuals and their households. They strongly suggest that economic class should not be analysed by itself, and that apparent class differences can be misinterpreted without gender analysis. Insufficient attention to intersectionality in much of the health literature has significant human costs, because those affected most negatively tend to be those who are poorest and most oppressed by gender and other forms of social inequality. The programme and policy costs are also likely to be high in terms of poorly functioning programmes, and ineffective poverty alleviation and social and health policies. PMID- 19288341 TI - Gender, health, and human rights in sites of political exclusion. AB - In this paper, we investigate the intersections of gender, health and human rights in sites of political exclusion. We apply the political theory of Giorgio Agamben on 'states of exception', seeking to better understand how the recent 'war on terror', that seemingly knows no limits of time or space, is driving health outcomes in refugee and Internally Displaced Persons (IDP) camps. Reproductive health, militarization, and gender-based violence in camps are explored in depth. The evidence presented reveals a number of contradictions of refugee and IDP camps, further highlighting the need for a more rights based humanitarianism. We conclude that foregrounding states of exception, as a way of understanding current gender dynamics in the social determinants of health, is both epidemiologically necessary and conceptually useful. We find that, in these sites of exclusion, the indispensability of a human rights approach to gender and health equity issues is revealed most directly. Furthermore, we are able to make new connections between the 'crisis of humanitarianism', gender, and health. PMID- 19288342 TI - Changing gendered norms about women and girls at the level of household and community: a review of the evidence. AB - Gendered norms are embedded in social structures, operating to restrict the rights, opportunities, and capabilities, of women and girls, causing significant burdens, discrimination, subordination, and exploitation. This review, developed for the Women and Gender Equity Knowledge Network of the WHO Commission on the Social Determinants of Health, sought to identify the best available research evidence about programmatic interventions, at the level of household and community, that have been effective for changing gender norms to increase the status of women. The focus was on developing countries. A wide range of single and multiple databases were searched, utilizing database specific keywords such as: women and girls; men and boys; household and community; intervention; and gender norms. Key themes were identified: education of women and girls; economic empowerment of women; violence against women, including female genital mutilation/cutting; and men and boys. Types of interventions, levels of action, populations of interest, and key outcomes from evaluations are identified. Evaluations are limited, with little evidence or measurement of changes in gender equity and women's empowerment. A key finding is, that targeting women and girls is a sound investment, but outcomes are dependent on integrated approaches and the protective umbrella of policy and legislative actions. PMID- 19288343 TI - Sex, gender, and vulnerability. AB - This paper is concerned with how sex chromosomes and gendered experience differentially contribute to health outcomes, and how gender effects provide an under-explored avenue for health intervention. Research on gender and health is currently undermined by conflation of sex and gender in much of the epidemiologic and clinical literature. This precludes any meaningful reflection on the extent to which our genetic blueprint, versus gendered socialization, contributes to the specific health vulnerabilities of males or females. Drawing on the 2002 global disability adjusted life years (DALYs) for males and females, this paper looks at health outcomes that differentially affect males and females, and distinguishes between vulnerabilities linked to the XX or XY genotype, vulnerabilities due to gendered life experience, and vulnerabilities about which we understand relatively little. The paper highlights the dynamic and changeable nature of gendered health vulnerabilities. Given that gender-based risks are, in principle, amenable to social change, they offer untapped potential for health interventions. PMID- 19288344 TI - Nurses, community health workers, and home carers: gendered human resources compensating for skewed health systems. AB - This review examines the experiences of nurses, community health workers, and home carers in health systems from a gender analysis. With respect to nursing, current discussions around delegation take place over layers of historical struggle that mark the evolution of nursing as a profession. Female community health workers also struggle to be recognized as skilled workers, in addition to defending at a personal level the legitimacy of their work, as it transgresses traditional norms proscribing morality and the place of women in society, at times with violent consequences. The review concludes by exploring the characteristics of, and challenges faced by, home carers, who fail to be recognized as workers at all. A key finding is that these mainly female frontline health workers compensate for the shortcomings of health systems through individual adjustments, at times to the detriment of their own health and livelihoods. So long as these shortcomings remain as private, individual concerns of women, rather than the collective responsibility of gender, requiring public acknowledgement and resolution, health systems will continue to function in a skewed manner, serving to replicate inequalities in the health labour force and in society more broadly. PMID- 19288345 TI - Gender biases and discrimination: a review of health care interpersonal interactions. AB - A good interpersonal relationship between a patient and provider, as characterized by mutual respect, openness, and a balance in their respective roles in decision-making, is an important marker of quality of care. This review is undertaken from a gender and health equity perspective and illustrates that gender biases and discrimination occur at many levels in the healthcare delivery environment, and affects the patient-provider interaction which can result in health inequities affecting individual health seeking behaviour, access to good quality healthcare, and, ultimately, health outcomes. Interventions will have to be introduced at multiple levels, from health system legislation and policy and gender sensitive training to the development of women and men centred services and health literacy programmes. PMID- 19288346 TI - Strengthening accountability to citizens on gender and health. AB - Accountability refers to the processes by which those with power in the health sector engage with, and are answerable to, those who make demands on it, and enforce disciplinary action on those in the health sector who do not perform effectively. This paper reviews the practice of accountability to citizens on gender and health, assesses gaps, and recommends strategies. Four kinds of accountability mechanisms have been used by citizens to press for accountability on gender and health. These include international human rights instruments, legislation, governance structures, and other tools, some of which are relevant to all public sector services, some to the health sector alone, some to gender issues alone, and some to gender-specific health concerns of women. However, there are few instances wherein private health sector and donors have been held accountable. Rarely have accountability processes reduced gender inequalities in health, or addressed 'low priority' gender-specific health needs of women. Accountability with respect to implementation and to marginalized groups has remained weak. This paper recommends that: (1) the four kinds of accountability mechanisms be extended to the private health sector and donors; (2) health accountability mechanisms be engendered, and gender accountability mechanisms be made health-specific; (3) resources be earmarked to enable government to respond to gender-specific health demands; (4) mechanisms for enforcement of such policies be improved; and (5) democratic spaces and participation of marginalized groups be strengthened. PMID- 19288347 TI - Gender mainstreaming in health: looking back, looking forward. AB - This paper reviews published literature on experiences in mainstreaming gender within the health sector since the 1990s. Although much has been written about the need for mainstreaming gender, and on how to go about it, the gap between intention and practice is palpable. National health policies and programmes that have gender integrally woven into their objectives and activities are rare. Health research to generate gender and sex-specific data, and integrating gender in health provider training, have received scarce attention. Mainstreaming gender within institutions has remained superficial, investing more on form than on content. The apparent lack of progress in mainstreaming gender in health may be attributed to: depoliticization and delinking of gender mainstreaming from social transformation and social justice agendas; adoption of top-down approaches to mainstreaming; growing hostility within the global policy environment to justice and equity concerns; and increasing privatization and retraction of the state's role in health. This paper suggests that the way forward would be to frame gender concerns in the language of equity, rights, and justice; to set agendas which consider gender inequity within the context of inequities by caste, class, ethnicity, and other sources of health inequalities; and to work alongside other movements for social justice. PMID- 19288348 TI - Policy briefing. Appendix. PMID- 19288349 TI - The contested politics of sexual and reproductive health and rights. PMID- 19288350 TI - Sexual politics and sexual rights in Brazil: an overview. AB - The objective of this text is to survey the political construction of sexual rights in Brazil working on themes that are especially relevant to the configuration of these rights (reproduction/abortion, STDs/AIDS and sexual diversity), and mapping the main actors, the legal instruments now in place or still being discussed and, lastly, the public policies that have been adopted in the last two decades. We considered the 1988 Brazilian Constitution the landmark from which, in terms of the civil society's perspective at least, demands for new rights are ordained and, at state level, public policies and legal instruments are generated to satisfy these demands. PMID- 19288351 TI - Political intersections between HIV/AIDS, sexuality and human rights: a history of resistance to the anti-sodomy law in India. AB - The HIV/AIDS epidemic in India has posed unprecedented challenges to both state and society, to question prevailing constructions of patriarchal gender relations and heteronormativity. Response to the challenge has come not from the political and social mainstream but from the criminalised "margins": people of alternative sexualities, who have launched a struggle for reform of the anti-sodomy law, Section 377 of the Indian Penal Code. This article documents the history of this movement, and identifies the multiple national and global-level cultural, political, and economic strands, shaping it. The legal reform movement has been invaluable as a tool to mobilise disparate alternative sexualities groups around a common strategy, thereby forging them into a tenuous national-level "community". Going beyond legal reform in the direction of sexual rights, however, requires a broader coalition of groups, and a broad-based political agenda of sexual rights for all. This agenda must critique patriarchy, dominant masculinity, and sexual violence; forces that together govern both the subordination of women and repression of alternative sexualities. PMID- 19288352 TI - Policies around sexual and reproductive health and rights in Peru: conflict, biases and silence. AB - This study is aimed at examining how subsequent Peruvian governments, since 1990, have addressed reproductive rights, HIV/AIDS prevention and treatment, and sexual diversity rights, as well as the drastic policy shifts and its many contradictions. Abortion and contraception consistently generated the deepest public controversies and debates, which made progress in reproductive rights difficult. HIV/AIDS was often portrayed as having the potential to affect everyone, which allowed advocates and activists to achieve some success in advancing HIV/AIDS-related rights. Sexual diversity rights, perceived as a demand made by "others", were generally trivialised and disdained by politicians, officials, and the general population. Positive changes occurred as long as the issue was given a low political and institutional profile. The analysis of policy making and programme implementation in these three areas reveals that: (1) Weaknesses in national institutional frameworks concerning reproductive health made it possible for governments to adopt two very different (even contradictory) approaches to the issue within the past 15 years; (2) Policies were presented as rights-based in order to garner political legitimacy when, in fact, they evidenced a clear disregard for the rights of individual citizens; and (3) By favouring low-profile "public health" discourses, and marginalising "the sexual" in official policies related to sexuality, advocacy groups sometimes created opportunities for legal changes but failed to challenge conservative powers opposing the recognition of sexual and reproductive rights and the full citizenship of women and sexual minorities. PMID- 19288353 TI - The social construction of ARVs in South Africa. AB - An estimated 5.5 million people are currently living with HIV/AIDS in South Africa, 4.9 million of them between the ages of 15-49, 18.8% of the total population in that age bracket (Department of Health, Republic of South Africa 2006). The potential medical and social benefits of anti-retroviral drugs (ARVs) would be substantial, but South Africa's leaders have faulted in their response to AIDS from the very beginning, particularly President Thabo Mbeki, who, in concert with the Minister of Health, has questioned the basic science of AIDS, and has condemned ARVs as poisonous. President Mbeki has created a false distinction between social causes and disease agents in his analysis that it is poverty, rather than HIV, that causes AIDS. He has made his arguments using post colonial rhetoric to condemn pharmaceutical imperialism and medical experimentation on African populations. Opponents, most notably the pro-treatment social movement group, Treatment Action Campaign, claim that because poverty increases the risk of infection, illness and death due to HIV access to anti retroviral medication is a social justice issue - justice demands the medications be available at all government clinics at no cost. In 2003 a government-sponsored treatment programme was launched, and by mid-2006 it was treating 140 000 persons with HIV/AIDS, less than 25% of the number estimated to require treatment. Treatment access, for all who need it in South Africa, is an ambitious but achievable goal. A new president will be elected in 2008, and many hope that this will result in a national treatment programme unshackled from the "AIDS denialism" of the current leaders. Former deputy president, Jacob Zuma, is likely to be the next president. His record on AIDS, and his patriarchal attitudes towards women, are troubling, however. One can only hope that the provincial health systems, which operate with a fair level of autonomy from the national Department of Health, will not be further hampered in their work by the politics of the central government. PMID- 19288354 TI - 'New bottle, but old wine': from family planning to HIV/AIDS in post-Doi Moi Vietnam. AB - This paper begins with an observation that during the past two decades HIV/AIDS has emerged as a new public health priority in Vietnam and has commanded increasing attention and resources from both the government and the donor community. By juxtaposing HIV/AIDS with family planning, another social and health priority that preceded, and overlapped with, HIV/AIDS for a large part of the 1990s, we show two major gaps that have undermined both programmes. One is the lack of a strong civil society, that could have served as advocates for change, especially outside the government and the donor community. The other is the desire for control of women's bodies and sexuality that has been driven by the ever shifting project of nation building. We argue that these two major gaps represent more continuity than discontinuity in the way sexual and reproductive health issues are approached in Vietnam despite the seeming shift in priority that the emergence of HIV/AIDS suggests. PMID- 19288355 TI - Looking for sex in all the wrong places. AB - The purpose of this paper is to present an analysis of how issues on sexuality are captured by the World Bank's economic rationality, producing a sanitised discourse which, through its silences, further contributes to a normalised view of sexuality. Given the Bank's authority to determine what kinds of health and development programmes are established in the developing world, it is in a unique position to influence approaches to issues of gender and sexuality. An analysis of the Bank's documents reveals, however, that rather than addressing these fundamental components of some of the most pressing health emergencies of our time, its economic rationality and technocratic viewpoint has effectively silenced and sanitised the discourse on sexuality, thereby limiting what sexuality and gender-related issues can be tackled in the context of Bank sponsored programmes, and constraining efforts to advance fundamental sexual rights. Nevertheless, unexpected and paradoxical results may arise from that process, which, thus, does not necessarily lead to the furthering of a comprehensive conservative agenda. PMID- 19288356 TI - Gaining access to Vietnam's cigarette market: British American Tobacco's strategy to enter 'a huge market which will become enormous'. AB - British American Tobacco (BAT) has made concerted efforts since the late 1980s to establish a major presence in Vietnam, among the world's 10 fastest growing tobacco markets. Until 2000, Vietnam's tight regulation of the industry has been largely driven by trade and investment policy, resulting in a stronger domestic industry but increased production and consumption of tobacco products. BAT gained market access, and achieved a dominant market share among TTCs, through leaf development, licensed manufacturing, and the contraband trade. With impending trade liberalization in Vietnam, the company is now well placed to further expand sales. The ambitious National Tobacco Control Policy, adopted in 2000, signals a shift in political priority towards the protection of public health. Effective implementation and enforcement of its comprehensive measures will depend on the public health community's ability to draw support from regional and global experience, notably the Framework Convention on Tobacco Control (FCTC). PMID- 19288357 TI - A common pathway toward women's health. AB - This paper calls for an alternate approach to studying the aetiology of women's health conditions. Instead of the long-established disease-specific, compartmentalized approach, it recommends focusing on risk exposures that allows for the identification of multiple disease conditions that stem from the same risk factors. Identifying common risk factors and the related pathways to adverse health outcomes can lead to the development of interventions that would favourably affect more than one disease condition. The utility of such an approach is illustrated by a review of literature from across the globe on the association between gender inequity-related exposures and women's health (namely, three health conditions: sexually transmitted infections [STIs], including Human Immunodeficiency Virus [HIV], blindness, and depression; as well as two risk behaviours: eating disorders and tobacco use). The review demonstrates how women's health cannot be viewed independently from the larger social, economic, and political context in which women are situated. Promoting women's health necessitates more comprehensive approaches, such as gender-sensitization of other family members, and the development of more creative and flexible mechanisms of healthcare delivery, that acknowledge the gender inequity-related constraints that women face in their daily lives. PMID- 19288358 TI - Northern Uganda and paradigms of HIV prevention: the need for social analysis. AB - In settings of armed conflict, traditional HIV prevention programmes that promote risk avoidance via abstinence and fidelity and risk reduction via condom use and needle exchange are not viable. In such contexts, HIV risk depends less on personal choice than on exposure to physical, emotional and structural violence. War in northern Uganda has created three realities (internally displaced people's camps, night commuters and child abductions) which increase vulnerability to HIV transmission. Based upon this analysis of northern Uganda, we offer a conceptual framework for HIV transmission in conflict settings that recognizes the importance of local and global context in creating vulnerability to HIV infection. This framework is then used to delineate strategies for HIV prevention in northern Uganda, namely the provision of a safe physical environment and access to education, medical and psychological support, and the promotion of conflict resolution strategies and human rights law. PMID- 19288359 TI - Building capacity and community resilience to HIV: a project designed, implemented, and evaluated by young Lao people. AB - A partnership was formed between a mass youth organization, a national HIV coordinating committee, and an international agency, to implement an HIV capacity building project in the youth sector of Lao PDR. Involving the local community in situation analysis, planning and skills-building was a key focus of the project. District project working teams were trained in situation analysis, strategic planning, proposal development, and the implementation of HIV prevention activities. Young village volunteers were trained in participatory research, analysis, and behaviour change communication to promote HIV prevention. After 6 years, the partnership used qualitative methods to evaluate the local outcomes of the project. We found that district project working teams and young volunteers had improved skills in the areas in which they had been trained. Communities and local government workers had developed greater understanding of the HIV situation in their districts, and expressed a strong sense of ownership over their activity plans. Young people more readily acknowledged personal risk of HIV infection and were more comfortable talking about sexually transmitted infections. Although there were challenges to sustaining project activities in some areas, we found that our approach helped to engage youth and build their resilience to HIV in this country of low prevalence. PMID- 19288360 TI - Prevention of mother-to-child transmission of HIV in a refugee camp setting in Tanzania. AB - The objective of this article is to describe the results of a 2-year pilot programme implementing prevention of mother to child HIV transmission (PMTCT) in a refugee camp setting. Interventions used were: community sensitization, trainings of healthcare workers, voluntary counselling and HIV testing (VCT), infant feeding, counselling, and administration of Nevirapine. Main outcome measures include: HIV testing acceptance rates, percentage of women receiving post test counselling, Nevirapine uptake, and HIV prevalence among pregnant women and their infants. Ninety-two percent of women (n=9,346) attending antenatal clinics accepted VCT. All women who were tested for HIV received their results and posttest counselling. The HIV prevalence rate among the population was 3.2%. The overall Nevirapine uptake in the camp was 97%. Over a third of women were repatriated before receiving Nevirapine. Only 14% of male counterparts accepted VCT. Due to repatriation, parent's refusal, and deaths, HIV results were available for only 15% of infants born to HIV-infected mothers. The PMTCT programme was successfully integrated into existing antenatal care services and was acceptable to the majority of pregnant women. The major challenges encountered during the implementation of this programme were repatriation of refugees before administration of Nevirapine, which made it difficult to measure the impact of the PMTCT programme. PMID- 19288361 TI - The origin of violent behaviour among child labourers in India. AB - We explored the causes and circumstances of violent behaviour among a group of child labourers in the Indian unorganized sectors. From 14 categories of occupations, a total of 1,400 child labourers were interviewed in both urban and rural areas. The average family size of these mostly illiterate child labourers is seven, and average family income is 3,200 INR per month. In the short term child labourers become violent, aggressive, and criminal, following a pyramid of violent behaviour, including socio-economic pressure, cultural deviance, and psychological pressure. When considering family history it seems that the problem is part of a vicious cycle of violence, which persists through generations and evolves with financial crisis, early marriage, and violence in the family and workplace. Our study demonstrates that the most vulnerable groups of child labourers belong to the following workplaces: dhabas, food stalls, rail/bus stations, rail-floor cleaning, and rag picking. Giving high priority to capacity building within the community, including support for locally-generated solutions, is warranted. PMID- 19288362 TI - The use of PDAs to collect baseline survey data: lessons learned from a pilot project in Bolivia. AB - We compared the use of personal digital assistants (PDAs) against the use of standard paper questionnaires for collecting survey data. The evaluation consisted of qualitative approaches to document the process of introducing PDAs. Fieldwork was carried out during June-July 2005 at 12 sites in Bolivia. Data collectors reacted positively to the use of the PDAs and noted the advantages and disadvantages of paper and PDA data collection. A number of difficulties encountered in the use of PDA technology serve as a warning for investigators planning its adoption. Problems included incompatible data files (which impeded the ability to interpret data), an inadequate back-up protocol, and lack of a good 'fit' between the technology and the study. Ensuring the existence of a back end database, developing an appropriate and adequate back-up protocol, and assessing whether a technology 'fits' the project are important factors in weighing the decision to collect data using PDAs. PMID- 19288365 TI - Partnership is a word often used, but a concept less often practiced. Foreword. PMID- 19288366 TI - An assessment of the positive partnership project in Thailand: key considerations for scaling-up microcredit loans for HIV-positive and negative pairs in other settings. AB - Stigmatization and discrimination against people living with HIV/AIDS (PLHA), and their families, remains a barrier to participation in prevention and care programmes. This barrier takes on added significance as Thailand expands provision of free antiretroviral therapy (ART). This paper documents an innovative approach to improve quality of life for PLHA, while reducing levels of stigma and discrimination. The Population and Community Development Association (PDA) began implementing the Positive Partnership Project (PPP) in 2002. In this project, an HIV-negative person must team up with an HIV-positive person to become eligible for a loan for income-generating activities. The use of microcredit to explicitly reduce stigma and discrimination is a unique feature of the PPP. While the microcredit component of the project is an important dimension for improving the status of participating PLHA, the impacts of the project extend far beyond the PLHA who receive loans. Both directly and indirectly, it has contributed to improved quality of life and economic conditions for PLHA, while raising their visibility and acceptance in hundreds of communities throughout urban and rural Thailand. This paper identifies key features of the project and considerations for adapting its use in other settings. PMID- 19288367 TI - University partnership to address the shortage of healthcare professionals in Africa. AB - The shortage of qualified health professionals is a major obstacle to achieving better health outcomes in many parts of the world, particularly in Africa. The role of health science universities in addressing this shortage is to provide quality education and continuing professional development opportunities for the healthcare workforce. Academic institutions in Africa, however, are also short of faculty and especially under-resourced. We describe the initial phase of an institutional partnership between the Muhimbili University of Health and Allied Sciences (MUHAS) and the University of California San Francisco (UCSF) centred on promoting medical education at MUHAS. The challenges facing the development of the partnership include the need: (1) for new funding mechanisms to provide long term support for institutional partnerships, and (2) for institutional change at UCSF and MUHAS to recognize and support faculty activities that are important to the partnership. The growing interest in global health worldwide offers opportunities to explore new academic partnerships. It is important that their development and implementation be documented and evaluated as well as for lessons to be shared. PMID- 19288368 TI - From pilot to mainstream: a decade of working in partnerships to expand access to emergency contraception in Mexico. AB - Emergency contraception (EC) is used post-coitally and offers women an important second chance to prevent pregnancy. Approximately 10 years ago, Mexico became one of the earliest countries working to expand EC access. Partners came together to pursue a synergistic four-pronged strategy, including product registration, health care provider training and orientation, awareness raising among the general public, and policy change. By 2006, five EC products were registered and sold in Mexico, EC was widely available through pharmacies and included in the public sector contraceptive options, and knowledge of the method had increased substantially. PMID- 19288369 TI - Multi-level partnerships to promote health services among internally displaced in eastern Burma. AB - Ethnic populations in eastern Burma are the target of military policies that result in forced labour, destruction of food supplies, and massive forced displacement. Despite international assistance to Burmese refugees along the Thai Burma border, traditional humanitarian models have failed to reach these internally displaced persons (IDPs) within Burma. Nevertheless, through the cultivation of a model (cross border local-global partnerships) 300,000 IDPs in eastern Burma now receive critical health services where, otherwise, there would be none. We describe key elements of the partnership model's genesis in eastern Burma. The role of the local partner, Backpack Health Worker Team (BPHWT), is highlighted for its indigenous access to the IDP populations and its maintenance of programmatic autonomy. These local elements are potentiated by international support for technical assistance, training, resources, and advocacy. International policy and investment should prioritize support of locally-driven health initiatives that utilize local-global partnerships to reach not only IDPs but also other war-torn or traditionally inaccessible populations worldwide. PMID- 19288370 TI - Health care at the end of the road: opportunities from 20 years of partnership in onchocerciasis control. AB - The unprecedented decision of Merck & Co., Inc., to donate ivermectin through the Mectizan(R) Donation Program, has catalysed an exemplary partnership, to distribute the drug to the communities at risk of onchocerciasis, and empower them to take charge of the drug distribution themselves. Integration with other activities has always been part of the plan, but has been accelerated in recent years because of the need to strengthen primary health care, and to meet the challenges of integrating the rapid impact of Neglected Tropical Disease programmes. Activities that have been integrated include provision of vitamin A capsules, elimination of lymphatic filariasis, the distribution of insecticide treated nets, and comprehensive eye health. Although these integrated activities show promising results for all programmes involved, challenges still remain. The risk of overburdening communities with multiple activities, and the problem of remuneration at the community level, are the major concerns, as is the need for effective coordination. The expanded onchocerciasis control partnership is a model of translating the eighth Millennium Development Goal (MDG), namely 'develop a global partnership for development', into action and also addresses other key MDGs. In 2006, the partnership provided more than 62 million treatments for onchocerciasis control, and offers a firm foundation from which to deliver other needed health interventions while safeguarding the achievements of onchocerciasis control thus far. PMID- 19288371 TI - Communication and social capital in the control of avian influenza: lessons from behaviour change experiences in the Mekong Region. AB - International development agencies, national governments, and nongovernmental organizations are increasingly collaborating with local civil society groups in mounting behaviour change communication (BCC) interventions. Even in countries with weakened civil societies, the social capital of local organizations can be a fundamental communication resource. The experience of three programmes in the Mekong Region that used BCC to prevent and control outbreaks of avian influenza bore out this finding. These programmes worked with the Vietnam Women's Union to mobilize local women as conduits for education; worked with the Centre d'Etude et de Developpement Agricole Cambodgien (CEDAC), in Cambodia, to educate and train village health promoters and model farmers; and worked with the Lao Journalists Association to educate and build skills among print and broadcast journalists to enhance avian influenza coverage. Collaborating with civil society organizations can enhance communication reach, trust, and local ownership, but poses many challenges, particularly institutional capacity. Our experience, nevertheless, holds promise for a measured approach that views social capital as a set of communication resources at the community level that can be mobilized to promote complex behaviours, particularly in a rapidly changing outbreak situation. PMID- 19288372 TI - Improving the management of hypertension in Kazakhstan: implications for improving clinical practice, patient behaviours and health outcomes. AB - This paper presents findings from the assessment of a strategy aimed at improving case-finding and management of hypertension patients. Study findings suggest that providers' orientation to clinical guidelines, public information on hypertension risks, promotion of yearly blood pressure screening, and universal access to an outpatient drug benefit package, improve case-finding and management. In addition, training of providers at pilot sites resulted in a substantial and significant difference in patient care and health outcomes. Provider training and tools, especially focused on effective patient counselling, made a significant contribution to increased case-finding, patient adherence to prescribed drugs, reducing salt, and increasing regular exercise. However, further refinements are required to achieve the expected adherence of patients to medication and lifestyle advice. In the pilot area, we compared two samples of patients before and after the initiation of the intervention. We found an increase in the proportion of patients with blood pressure <140/90 mmHg, and a significant decrease in the proportion of patients with blood pressure > or =160/100 mmHg. The strategy involved the joint efforts of the Research Institute of Cardiology and Internal Diseases, Karaganda Drug Information Centre, Karaganda Oblast Health Department, Kazakhstan Association of Family Physicians, Almaty Postgraduate Institute for Physicians, and USAID ZdravPlus Project in Central Asia. PMID- 19288373 TI - Management and mortality in patients with non-ST-segment elevation vs. ST-segment elevation myocardial infarction. Data from the Malopolska Registry of Acute Coronary Syndromes. AB - BACKGROUND: According to the presenting electrocardiogram, acute myocardial infarction (MI) can by categorised generally as non-ST-segment elevation MI (NSTEMI) and ST-segment elevation MI (STEMI). AIM: To assess the impact of the different acute MI categories on in-hospital management and mortality in hospitals without on-site invasive facilities. METHODS: We identified 380 NSTEMI and 334 STEMI patients treated in the Malopolska Registry of Acute Coronary Syndromes from February to March 2005 and from December 2005 to January 2006. Data concerning in-hospital management and mortality were assessed. RESULTS: Patients with NSTEMI were older and were more likely to have prior angina, prior MI and prior heart failure symptoms than STEMI patients. The NSTEMI patients were less likely to be transferred for invasive treatment (23.9 vs. 41.9%, p <0.0001) and receive glycoprotein IIb/IIIa inhibitors during index hospital stay. The use of low-molecular-weight heparin, beta-blockers, angiotensin-converting enzyme inhibitors/angiotensin II antagonists, nitrates and statins was more frequent in NSTEMI patients. Among patients treated non-invasively, in-hospital mortality was high, but was lower in NSTEMI than STEMI patients (12.1 vs. 22.7%, p <0.0001). Independent predictors of in-hospital death in this group were age, cardiogenic shock, chronic obstructive pulmonary disease, and STEMI. CONCLUSIONS: Despite current recommendations, NSTEMI patients are still less likely to be transferred for invasive treatment than STEMI patients. Among patients treated non-invasively during index hospital stay, NSTEMI is associated with more favourable prognosis than STEMI, but the risk of in-hospital death is high. The hospital network should implement more frequently the strategy of early and urgent invasive treatment of NSTEMI patients. PMID- 19288374 TI - Outcomes of a single centre registry of patients with ischaemic heart disease, qualified for an RF ablation of ventricular arrhythmia after ICD intervention. AB - BACKGROUND AND AIM: Reduction of ICD interventions improves the quality of life and possibly reduces mortality. Ablation reduces ICD interventions in patients with ablatable arrhythmia, but its effectiveness needs to be proven for patients with coronary artery disease (CAD) regardless of the type of arrhythmia. Our study was designed to address this issue, but it had to be terminated due to recruitment problems. The reasons for early termination are described in this paper. METHODS: Patients with CAD and implanted ICD, who within the past three months survived an episode of VT/VF, were selected for this study. Patients were to be randomised for ablation or pharmacotherapy. A group of 209 patients was screened between June and December of 2007. RESULTS: Out of 209 patients, 39 (18.7%) had appropriate ICD therapy during the last three months and were potentially eligible for the trial. Out of 39 patients, 34 could not be randomised, due to the presence of exclusion criteria (n-25) or consent refusal (n-9). Previous ablation (n-10), left ventricular thrombus (n-3) or presence of mitral or aortic artificial valve (n-3) were the most frequent exclusion criteria. During follow-up of 12 months one patient required ablation due to frequent ICD discharges. From the five randomised patients, two were randomised to ablation and three to the pharmacotherapy arm. CONCLUSIONS: 1. Ablation might not be suitable as a routine treatment for all patients with ICD interventions, as a significant group prefers not to undergo RF ablation as a routine treatment or there are contraindications for the ablation. 2. There are obstacles in prospective and randomised evaluation of the role of ablation in patients with CAD and ICD interventions, which are related both to patients' medical conditions and to patients' will. These limitations should be taken into account when designing further studies. PMID- 19288375 TI - Changes in heart rate variability caused by coronary angioplasty depend on the localisation of coronary lesions. AB - BACKGROUND: Coronary angioplasty (PTCA) is a common treatment method in patients with coronary heart disease, but its effects on heart rate variability (HRV) have not been well established. AIM: To verify whether the localisation of coronary lesion undergoing PTCA affects HRV parameters. METHODS: Ninety six consecutive individuals underwent elective coronary angiography with subsequent ad hoc successful PTCA. Two five-minute ECG were recorded, one before PTCA and the second 24-hour after PTCA. The HRV indices were determined by means of classical and 'new' mathematical models. RESULTS: The PTCA-induced changes in HRV variables depended on the localisation of dilated lesion. PTCA of the circumflex artery revealed the most significant HRV changes--a decrease in value of domain indices: Yeh DI (0.033+/-0.031 vs. 0.011+/-0.006 un/unitless, p=0.005), Yeh II (0.053+/ 0.039 vs. 0.032+/-0.013 un, p=0.017), Organ BAND (9.101+/-9.245 vs. 4.62+/-2.205 bpm/beat per minute, p=0.031), Huey STV (208.821+/-262.248 vs. 76.444+/-35.281 bpm, p=0.013), Dalton MABB (15.733+/-16.575 vs. 7.57+/-4.89 ms, p=0.015), Dalton SD (48.741+/-37.468 vs. 27.759+/-10.533 ms, p=0.015), Zugaib STV (0.0129+/-0.0132 vs. 0.005+/-0.003 un, p=0.005), SDNN (27.204+/-18.592 vs. 21.329+/-32.784 ms, p=0.044), rMSSD (56.239+/-19.751 vs. 51.496+/-43.889 ms, p=0.025) and increased LF/HF (2.384+/-2.072 vs. 5.632+/-5.379 un, p=0.044). Angioplasty of the right coronary artery resulted in decreased AR TP (18.273+/-2.296 vs. 17.085+/-2.256 ms(2), p=0.017) and alteration of the sympathovagal balance of the autonomic nervous system towards predominance of sympathetic activity: AR LF (0.264+/-0.029 vs. 0.284+/-0.040 un, p=0.007), LF/HF (4.310+/-4.457 vs. 6.958+/-7.013 un, p=0.018), HF (0.199+/-0.165 vs. 0.141+/-0.157 un, p=0.031), AR HF (0.647+/-0.043 vs. 0.621+/-0.054 un, p=0.014). PTCA of the left anterior descending artery caused no change. CONCLUSION: Changes in heart rate variability caused by coronary angioplasty depend on the localisation of coronary lesions. PMID- 19288376 TI - Factors predicting discontinuation of a hospital-based cardiac rehabilitation programme. AB - BACKGROUND: Cardiac rehabilitation (CR) programmes play an important role in the management of patients with coronary artery disease. However, a significant proportion of patients do not participate or do not complete CR. AIM: To asses the prevalence and predictors of discontinuation of a hospital-based CR programme and to investigate whether or not the completers and dropouts differed in relation to their baseline characteristics. METHODS: Data used for analysis were from a hospital-based CR programme involving 1986 discharged patients at Tehran Heart Centre between July 2004 and January 2006. The patients who completed all 24 sessions of the CR programme were compared with the dropouts. RESULTS: The CR completion rate was 18.1% (average of 11.4+/-8.1, ranging from 1 to 78 sessions) including patients who completed 24 (n=284) or more sessions (n=77) of the CR programme. Factors predicting dropout were male gender (OR 1.441, p=0.0094), younger age (OR 0.979, p=0.005), and lower levels of education (OR 0.412, p <0.0001). CONCLUSION: The present study demonstrated a relatively high rate of CR programme dropout. Only less than a fifth of the patients completed this hospital based programme. Patients who were male and younger and had lower education levels were better likely to drop out of the CR programme. Social support and educational programmes may be helpful in achieving better compliance. PMID- 19288377 TI - Percutaneous extraction of endocardial leads--a single centre experience in 120 patients. AB - BACKGROUND: The expanding number of patients treated with pacing, especially resynchronisation therapy and pacing system upgrades as well as leads remaining inactive, and prolonged life expectancy are the causes of an increase of the number of electrodes in pacemaker patients. The growing problem with endocardial lead infections and excess leads has made percutaneous lead removal technology widespread as it is less invasive than cardiosurgery. AIM: We present our experience in percutaneous lead removal in a single reference centre in Poland. METHODS: During 2.5 years, 236 leads in 120 patients were removed. The criterion for inclusion in the present analysis was the age of the oldest lead: > 12 months in pacemaker patients and > 6 months in patients with implantable cardioverter defibrillators (ICD). All patients admitted to the hospital for lead removal underwent a percutaneous procedure. The age of the patients ranged from 18 to 87 (mean 65.7) years. The leads were removed using the Lead Extraction System (Cook) with the rotational cutting force only, notlaser or RF energy. RESULTS: Indications for lead removal were: local (pocket) infection (47%), endocarditis (27%) and lead excess (26%). Seventy six percent of patients had at least two pacemaker/ICD-related procedures whereas 24% had only one implantation procedure in the past. The median time from the preceding procedure was 12 months. In 38 patients there were 60 inactive electrodes. The majority of patients had two (62%) or three (19%) leads, followed by 12% with one lead and 7% of patients with more than three leads. In 27.5% of patients leads from the coronary sinus were removed. The complication rate was 4%. CONCLUSIONS: Percutaneous lead removal procedures are performed in Poland for class I and II indications according to NASPE classification. In many cases patients had multiple leads, including in the coronary sinus. The majority of patients had two or more interventions in the past. PMID- 19288378 TI - [Brugada syndrome--underestimated cause of sudden cardiac death in patients without an organic cardiac disease--a case report]. AB - We present a case of a 61-year-old man with a history of syncope and sudden cardiac arrest due to ventricular fibrillation, with the symptoms of spontaneous electrocardiographic changes characteristic for the Brugada syndrome. The patient received an implantable cardioverter-defibrillator. This patient was previously hospitalised in 2003, when the diagnosis of atrio-ventricular block has been established and in 2006, when he had chest pain and was operated because of the right coronary artery ostium haematoma. However, the diagnosis of Brugada syndrome was not established. PMID- 19288379 TI - [Myocardial infarction and pulmonary embolism in a 45 year-old woman with the prothrombin 20210A mutation--a case report]. AB - We present a case of a 45-year-old-female who developed myocardial infarction and 6 weeks following first hospitalisation--pulmonary embolism. The disease was probably triggered by hormone replacement therapy. Coronary angioplasty and anticoagulant therapy with unfractioned heparin and acenokumarol were used with success. The woman and her 17-year-old daughter were shown to be heterozygous for prothrombin mutation G20210A. This case shows that trombophilia should be considered in young women with deep venous thrombosis, pulmonary embolism, venous thromboembolism or with myocardial infarction especially in women with high-risk factors. PMID- 19288380 TI - Acute onset of chronic aortic dissection presenting as abdominal pain. AB - A 54-year-old male patient developed acute onset of chronic aortic dissection manifesting as primarily abdominal pain. A prompt diagnosis was made and urgent surgery was carried out successfully under profound hypothermic circulatory arrest. Prompt surgery is mandatory for such patients in order to prevent abrupt rupture. The awareness of possible aortic dissection is the key point leading to an early diagnosis in patients with atypical presentations. PMID- 19288381 TI - [Acute myocardial infarction in a patient with acute ischemic stroke and angiographically normal coronary arteries]. AB - Myocardial infarction with angiographically normal coronary arteries is a puzzling clinical condition described since the implementation of coronary angiography as a diagnostic technique. We report a case of a 55-year-old male patient with acute myocardial infarction and acute ischemic stroke with normal coronary arteries. Diagnostic dilemmas and problems with correct treatment are discussed. PMID- 19288382 TI - Haissaguerre syndrome--a new clinical entity in the spectrum of primary electrical diseases? PMID- 19288384 TI - [Recanalisation of chronic total occlusions--why, when and how?]. PMID- 19288385 TI - [ECG changes in patients with stroke]. AB - We present a case of a male patient who was admitted to the cardiology department with a diagnosis of NSTEMI (deep negative T waves in all precordial leads). Two weeks earlier he was hospitalised because of a stroke caused by thrombosis of the left carotid artery. He had no angina. An ECHO showed a normal myocardial contractility of the left ventricle. The concentration of troponin I was also normal. No coronary artery stenosis was revealed in coronary angiography. We present a review of studies devoted to ECG changes in patients with stroke. PMID- 19288386 TI - [The floating structure connected with endocavitary electrode--thrombus or bacterial vegetation?]. AB - A case of a 52-year-old woman with complete atrioventricular block, treated with pacemaker implantation, is presented. During the control transthoracic echo exam we observed floating structure connected with the endocavitary electrode. After diagnostic examinations bacterial vegetation was excluded and thrombus was diagnosed. The therapy with unfractionated heparin and warfarin was successfully performed. PMID- 19288387 TI - [A myocardial infarction in a patient with numerous coronary artery aneurysms]. AB - We present a case of a 53-year-old smoking patient with myocardial infarction, in whom coronary angiography revealed many coronary arteries aneurysms, without substantial epicardial vessel narrowings. Patient was medically treated with a good outcome. PMID- 19288388 TI - [Incessant septal tachycardia eliminated by RF applications from right and left atrium]. AB - We present a case of a 29-year-old woman who suffered from incessant atrial tachycardia (AT) leading to tachycardia-induced cardiomyopathy. Based on classical mapping of the right and left atria the focus of AT was localised in the inter-atrial septum area. The RF applications from the right atrium stopped AT for some seconds. Applications from the left atrium terminated AT definitively. The patient has been free from symptoms of arrhythmia and heart failure for seven years. PMID- 19288389 TI - [First in Poland transcatheter, transfemoral aortic valve implantation in elderly symptomatic high-risk patient with aortic stenosis--novel Zabrze experience]. PMID- 19288390 TI - Body composition of female wheelchair athletes. AB - Wheelchair users undergo changes in body composition as a result of disability. In this study the distribution of bone mineral, lean and fat mass was assessed in highly-trained female wheelchair athletes and a reference group by dual-energy X ray absorptiometry (DXA). The transferability of anthropometric equations commonly used in female groups was examined in order to establish a suitable field method of body composition assessment. The DXA total-body results indicated no difference between groups, but segmental analyses uncovered regional differences. The wheelchair athletes had greater BMD (p=0.088), more lean mass (p<0.001) and a lower percent fat (p=0.050) in their arms. The reverse was true of the legs (p< or =0.001). The trunk as a whole did not differ between groups. In general, the anthropometric equations showed a lack of transferability to the wheelchair group and tended to underestimate total percent body fat. Anthropometric measures such as body mass index (BMI) and waist girth showed strong correlations with body fat in the wheelchair group (BMI: r=0.90, p=0.001; waist: r=0.83, p=0.001), but weaker results in the reference group. It is recommended that specific anthropometric equations be developed for use in the absence of a 'gold standard' measure of body composition such as DXA. PMID- 19288391 TI - Sweat rate and sweat electrolyte composition in international female soccer players during game specific training. AB - The purpose of this study was to determine the sweat rate and sweat electrolyte composition in female international level soccer players. Thirteen soccer players performed two 90 min soccer-specific training sessions (T1 and T2) on separate days. Hydration status was determined prior to each session and sweat loss, sweat rate and sweat composition (Na (+), K (+), Mg (++) and Ca (+)) were determined from patches worn during training. The mean sweat rate during T1 and T2 was 0.50+/-0.20 and 0.43+/-0.18 L.h (-1) respectively (P>0.05). The mean sweat electrolyte composition during T1 and T2 was: [Na (+)]: 43.9+/-15.0 and 46.2+/ 7.9 mmol.L (-1); [K (+)]: 6.1+/-1.1 and 5.2+/-1.1 mmol.L (-1); [Mg (++)]: 0.1+/ 0.0 and 0.1+/-0.0 mmol.L (-1); [Ca (+)]: 1.2+/-0.5 and 0.7+/-0.1 mmol.L (-1), respectively. When data from T1 and T2 were combined, there were no relationships between sweat rate and sweat concentration of any electrolyte. In conclusion, the sweat rate and sweat electrolyte losses in this cohort of international female soccer players, during soccer-specific training in cool conditions, were small. Electrolyte losses of this magnitude are unlikely to require special consideration in terms of optimising player hydration. PMID- 19288392 TI - Type of exercise alters VE/VCO2slope in CAD patients. AB - The exercise carbon dioxide equivalent slope predicts prognosis in coronary artery disease patients. However, no study examined whether the carbon dioxide equivalent slope in coronary artery disease patients is dependent on the type of exercise. Twenty-nine coronary artery disease patients performed maximal cardiopulmonary exercise tests on bike, treadmill and arm cycle, with assessment of oxygen uptake, carbon dioxide output, and expiratory volume. The carbon dioxide equivalent slope was calculated from rest till peak exercise. The carbon dioxide equivalent slope was significantly different between walking, cycling, and arm cranking exercises (P<0.05), i.e. the carbon dioxide equivalent slope was the lowest during walking and the highest during arm cranking. Bland-Altman plots revealed that the difference of the carbon dioxide equivalent slope between exercises was greater in the case of a higher averaged carbon dioxide equivalent slope and overall peak oxygen uptake. In conclusion, the carbon dioxide equivalent slope is significantly dependent on the type of exercise in coronary artery disease patients. Additionally, the averaged carbon dioxide equivalent slope and peak oxygen uptake affect the difference of the carbon dioxide equivalent slopes between exercises. PMID- 19288393 TI - [Measuring life satisfaction in children and adolescents--psychometric properties of the "Questions on Life Satisfaction" FLZ(KJ)]. AB - OBJECTIVES: Measuring life satisfaction in children is becoming more and more important in clinical practice. This is the first study to evaluate the psychometric properties of the "Questions on Life Satisfaction for Children and Adolescents" FLZ(KJ) based on a student sample. METHODS: 1 312 German students ( M=13.7 years, SD=1.4) filled in the FLZ(KJ). Item characteristics, reliability, convergent and discriminant validity were assessed. Age- and gender-specific reference data were reported. RESULTS: Items were well accepted by the students. Selectivity and internal consistency of the two modules were acceptable, with alpha scores of alpha=0.70 and 0.75. Associations with the KIDSCREEN-10 Index support the convergent valid-ity of the instrument. The FLZ(KJ) differentiated well between children with and without chronic conditions. CONCLUSIONS: The FLZ(KJ) are reliable and sufficiently valid, with psychometric properties comparable to the adult version of the instrument. The instrument is a useful tool in measuring life satisfaction in children and adolescents. PMID- 19288394 TI - Wernicke's encephalopathy complicating pregnancy in a woman with neonatal necrotizing enterocolitis and resultant chronic malabsorption. AB - Wernicke's encephalopathy (WE) is a metabolic disturbance resulting from severe thiamine deficiency classically described in malnourished alcoholics. Untreated, it can result in stupor, coma, and death. WE has previously been reported as a complication of pregnancy in women with hyperemesis gravidarum. We report a case of WE complicating pregnancy in a woman with chronic malabsorption secondary to premature birth and subsequent necrotizing enterocolitis (NEC). Our patient progressed through classic stages of WE before lapsing into a coma. She made a complete recovery after aggressive intravenous thiamine and nutritional support. This is the first report of WE in pregnancy secondary to NEC-related chronic malabsorption. We report this case to bring attention to a potential pregnancy complication affecting women with malabsorptive conditions. PMID- 19288395 TI - Association of uterine artery Doppler resistance index and birth weight: effect of customized birth weight standards. AB - We assessed the effect of using customized birth-weight standards on the association of uterine artery Doppler resistance index (RI) and birth weight. This was a cross-sectional study of 2035 pregnancies scanned at 19 to 23 weeks. Both uterine arteries were identified using color flow mapping, and the RI was measured. The pregnancy outcome was obtained from the delivery suite database and the customized birth-weight centile calculated for each pregnancy. Both birth weight and customized birth-weight centiles were converted to z-scores. The correlation of uterine artery RI with birth weight was examined using conventional and customized birth-weight standards. Conventional and customized birth weights showed a significant correlation with uterine artery RI ( R = 0.062 and -0.208, respectively). The correlation was significantly improved by the use of customized birth weight. Use of customized birth weight significantly improves the correlation of uterine artery RI and birth weight. This finding may partly explain the lower sensitivity of uterine artery Doppler screening for fetal growth restriction as compared with preeclampsia. PMID- 19288396 TI - Pregnancy and genital sarcoma: a systematic review of the literature. AB - We conducted a literature review to determine the clinical characteristics of genital sarcoma during pregnancy. The systematic literature search was conducted using the search engines PubMed and MEDLINE with keywords "sarcoma" and "pregnancy" and was limited to female genital organs such as ovary, uterus, cervix, vagina, vulva, and retroperitoneal sarcoma. Kaposi's sarcoma, metastatic sarcoma, history of sarcoma, bone sarcoma located in pelvis, and fetal sarcoma were excluded in this study. There were 40 cases of genital sarcoma during pregnancy between 1955 and 2007. The majority of the cases were uterine sarcoma (37.5%), followed by retroperitoneal sarcoma (27.5%), vulvar sarcoma (22.5%), and vaginal sarcoma (12.5%). Mean age of the patient was 27.8 +/- 7.0. The distribution in the onset of symptoms had two peaks: first trimester (27.5%) and third trimester (50.0%). Growing mass (42.5%), abdominal pain (30.0%), and vaginal bleeding (22.5%) were the three most common symptoms. Incidental diagnosis was made in 22.5% and included during cesarean section (12.5%) and routine pelvic exam (7.5%). The cases initially not suspicious for malignancy were 42.5%. Thirty-three (82.5%) cases had live-born infants with term delivery in 55.2%. Mean birth weight was 2843 +/- 791 g, and male infants were more common (66.7%). Intrauterine growth retardation was seen in 12.5% of cases. Preterm labor was a common complication. Median survival period was 2.5 years (95% confidence, 1.9 to 3.1). The 2-, 3-, and 5-year cumulative survival rates were 60%, 38%, and 17%, respectively. Genital sarcomas in pregnancy are rare. There is a delay in diagnosis due to low index of suspicion. A majority had live births, and the 5-year survival is similar to that of advanced-stage sarcoma in nonpregnant women. PMID- 19288397 TI - Decreased first-trimester maternal serum free-beta subunit human chorionic gonadotropin and preterm birth in twin gestations. AB - We investigated whether abnormal concentrations of first-trimester free-beta subunit human chorionic gonadotropin (fsshCG) and pregnancy-associated plasma protein A (PAPP-A) are associated with preterm delivery in twin gestations. This was a hospital-based, retrospective study of 70 twin gestations between 11 (1)/ (7) and 13 (6)/ (7) weeks' gestation undergoing first-trimester screening. Free betahCG and PAPP-A multiples of the median were determined by our laboratory standards. Odds ratios (ORs) were estimated that compared the prevalence of very preterm (< 32 weeks), preterm (> or = 32 and < 37 weeks), and term birth (> or = 37 weeks) between the lower and higher percentile groups for each analyte. FsshCG levels < or = 25th percentile were associated with very preterm birth < 32 weeks' gestation (OR 5.10; 95% confidence interval [CI]: 1.19 to 21.95), but not with preterm birth > or = 32 and < 37 weeks' gestation (OR 0.50; 95% CI: 0.16 to 1.61). PAPP-A was not associated with very preterm (OR 2.95; 95% CI: 0.69 to 12.60) or preterm birth (OR 0.71; 95% CI: 0.23 to 2.21). Low first-trimester fsshCG was a strong predictor for very preterm birth in twin gestations. Low first-trimester PAPP-A was associated with a trend in increased risk of very preterm birth. PMID- 19288398 TI - Expectant management of severe preeclampsia at 27(0/7) to 33(6/7) weeks' gestation: maternal and perinatal outcomes according to gestational age by weeks at onset of expectant management. AB - We sought to determine perinatal outcome and maternal morbidities based on gestational age (GA) at onset of expectant management in severe preeclampsia (PE) between 27(0/7) and 33 (6/7) weeks. In this retrospective analysis of outcome in patients with severe preeclampsia, we studied 66 patients (71 fetuses) with severe PE at 27 (0)/ (7) to 33(6/ 7). All patients received corticosteroids. Perinatal and maternal complications were analyzed. Five patients had twin gestations. Median for days of prolongation was 5 days (range, 3 to 35). Birth weights of 19 (27%) were < 10% for gestational age, and 6 (8%) were < 5%. All fetuses survived except for one neonatal death at 27 weeks, and three infants had chronic lung disease-two at 27 and one at 28 weeks-but there were no cases of intraventricular hemorrhage (>or= grade ?). Rate of abruption was significantly higher at 27 to 28 weeks as compared with > 28 weeks (25% vs 6%, p = 0.05). There was no eclampsia, and two had transient renal insufficiency at 27 weeks. Four of 11 (36%) patients with expectant management at >or= 32 weeks had pulmonary edema or hemolytic anemia, elevated liver enzymes, and low platelet count. During expectant management, rate of respiratory distress syndrome and other serious neonatal complications decrease with increasing GA, supporting a role for such management in early severe preeclampsia. Because there is significant maternal morbidity at >or= 32 weeks with minimal neonatal benefit, consideration should be given for delivery of these pregnancies following corticosteroid administration. PMID- 19288399 TI - [Update of guidelines for patient self-management of oral anticoagulation]. PMID- 19288400 TI - Quality control of herbal material and phytopharmaceuticals with MS and NMR based metabolic fingerprinting. AB - Metabolic fingerprinting techniques have received a lot of attention in recent years and the annual amount of publications in this field has increased significantly over the past decade. This increase in publications is due to improvements in the analytical performance, most notably in the field of NMR and MS analysis, and the increased awareness of the different applications of this growing field. Metabolomic fingerprinting or profiling is continuously being applied to new areas of research such as drug discovery from natural resources, quality control of herbal material, and discovering lead compounds. In this review the current state of the art of metabolic fingerprinting, focussing on NMR and MS technologies will be discussed. The application of these two analytical tools in the quality control of herbal material and phytopharmaceuticals forms the major part of this review. Finally we will look at the future developments and perspectives of these two technologies in the quality control of herbal material. PMID- 19288401 TI - Effect of seven tricyclic diterpenoids from needles of Taxus media var. Hicksii on stimulus-induced superoxide generation, tyrosyl or serine/threonine phosphorylation and translocation of cytosolic compounds to the cell membrane in human neutrophils. AB - Taxol has been widely used as an anticancer drug for ovarian, breast, lung and prostate cancer. Some kinds of Taxus plants are widely distributed in the Northeast Asia region. We have isolated seven tricyclic diterpenoids, taxinine, taxagifine, 5-O-cinnamoyltaxacin I triacetate, 5-decinnamoyltaxinine J, 5 cinnamoyl-9-acetyltaxicin I, taxacin and taxol from the needles of Taxus media var. Hicksii, and investigated their effects on stimulus-induced superoxide generation and translocation of cytosolic compounds to the cell membrane in human neutrophils. Six tricyclic diterpenoids used in this experiment suppressed the superoxide generation induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) and arachidonic acid (AA) in a concentration-dependent manner. Taxinine significantly suppressed the superoxide generation induced by phorbol 12 myristate 13-acetate (PMA). The compounds also suppressed fMLP- and AA-induced tyrosyl or PMA-induced serine/threonine phosphorylation, and translocation of cytosolic compounds, p47 (phox), p67 (phox) and Rac to the cell membrane in parallel with the suppression of the stimulus-induced superoxide generation. PMID- 19288402 TI - Influence of antivitamins ginkgotoxin 5'-phosphate and deoxypyridoxine 5' phosphate on human pyridoxine 5'-phosphate oxidase. AB - The pharmacological effects of leaf extracts (EGb 761) from Ginkgo biloba L. are attributed to ginkgolides, bilobalide and biflavonoids. However, besides these beneficial attributes, ginkgotoxin, a B(6) antivitamin which may cause epileptic convulsions, other severe neuronal disorders and even death, is also found in Ginkgo leaves and leaf-derived remedies. Because of its structural similarity to the B(6) vitamers, an interaction of ginkgotoxin with enzymes involved in the vitamin B(6)-dependent metabolism of the human brain is possible. This led us to investigate how the neurotoxic ginkgotoxin acts in the brain. To this end the gene coding for the human pyridoxine 5'-phosphate oxidase was heterologously overexpressed in E. COLI and the homogeneous enzyme was characterized. The investigation showed that the enzyme is inhibited in vitro by the synthetic vitamin B(6) derivative 4'-deoxypyridoxine 5'-phosphate but not by ginkgotoxin or its 5'-phosphate. PMID- 19288403 TI - Comparative anticancer and antioxidant activities of different ingredients of Ginkgo biloba extract (EGb 761). AB - Flavonoid glycosides are the major constituents of Ginkgo biloba extract (EGb 761) and are well known to be an antioxidant for inhibiting tumor growth. Because it contains several flavonoid glycosides and other bioactive substances, the activities of EGb 761 against cancer in vivo and in vitro remain poorly understood. This situation prompted interest in the compounds for experimental study. We have investigated the anticancer effects of three analogues of EGb 761 samples on sarcoma 108 (S180)-bearing mice and leukemic 1210 (L1210) cell lines. We have also evaluated the changes of endogeneous antioxidant scavenging enzymes, including superoxide dismutase (SOD), glutathione (GST), lipid peroxidation (LPx), and catalase (CAT), in the blood of the S180-bearing mice. The EGb 761, EGb 761-H (containing mainly flavonoid aglycones and terpene trilactones), and EGb 761-DT-H (containing mainly flavonoid aglycones) samples exhibited cytotoxicity and inhibitory activity with IC (50) values of 46.36 +/- 2.43 microM, 10.27 +/- 0.88 microM, and 14.93 +/- 0.73 microM in L1210 cell-based assays, respectively. This resulted in 41.74 %, 60.72 %, and 63.76 % reductions in tumor weight after 10 days of treatment, respectively. In summary, the anticancer activity of EGb 761 can be improved by increasing the concentration of the aglycone form of the flavonoid. Terpene trilactones cannot exert the anticancer effects of flavonoids in vivo. Raising the levels of the free radical scavenger enzymes GST, SOD and CAT may be one of the involved anticancer mechanisms. PMID- 19288404 TI - Influence of sample preparation on the assay of isoflavones. AB - The complexity of sample matrices, coexistence of multiple forms of bioactive phytochemicals, and their interaction of with other cellular components pose a significant challenge for optimized extraction and accurate estimation of bioactive phytochemicals in foods and dietary supplements. This article discusses the significance of optimizing extraction procedures for accurate assay of phytochemicals from different matrices using bioactive isoflavones as model substrate because isoflavones are known to exist in nature as free aglycones or as conjugates with sugars and/or acids. The wide structural diversity and polarities of free and conjugated isoflavones makes optimum extraction and accurate quantification of isoflavones a challenging task. This paper reviews variables, extraction solvent composition (aqueous-organic solvents mixtures at different acidification levels), physical extraction techniques (Soxhlet, stirring, ultrasonic, microwave, pressurized, supercritical-fluid, high-speed counter-current chromatography), and parameters (temperature, pressure, number of cycles, solid-solvent ratio) that influence quantitative extraction of isoflavones from different matrices. In addition, this review covers a brief overview of structures, sources, bioactivities, separation, and detection used for isoflavones analysis. Optimum extraction efficiencies of isoflavones were obtained with EtOH : H (2)O : DMSO (70 : 25 : 5, v/v/v) as the extraction solvent and acidification of extraction solvent favored partial degradation of conjugated forms to their corresponding aglycones. Accurate quantification of isoflavones in foods, plants, and dietary supplements will allow researchers and regulators to provide more precise guidelines on dietary intake and safety levels necessary to achieve optimum health. PMID- 19288405 TI - 15-Lipoxygenase inhibitory effects of prenylated flavonoids from Erythrina senegalensis. AB - One new pterocarpan (named erybraedin F) along with seven known prenylated flavonoids were isolated from the stem and root bark of the medicinal plant Erythrina senegalensis. The structures were elucidated by 1D and 2D NMR and MS. The pterocarpans were found to be strong inhibitors of 15-lipoxygenase. PMID- 19288406 TI - Cytotoxic properties of root extract and fruit juice of Trichosanthes cucumerina. AB - The root extract of Trichosanthes cucumerina L. and bryonolic acid (1), its main constituent, as well as the fruit juice and cucurbitacin B (3), its main constituent, were tested for cytotoxicity against four human breast cancer cell lines (SKBR3, MCF7, T47D, and MDA-MB435), two lung cancer cell lines (A549 and SK LU1), and one colon cancer cell line (Caco-2). The root extract had higher IC (50) values than bryonolic acid (1) against three breast cancer cell lines (MCF7 = 267/121, T47D = 316/124, MDA-MB435 = 140/90 microL/mL) and one lung cancer cell line (A549 = 106/100 microL/mL). The fruit juice also had higher IC (50) values than cucurbitacin B (3) against the four breast cancer cell lines (131/73, 375/35, 249/60, and 156/26 microL/mL, respectively) and one lung cancer cell line (141/41 microL/mL) as shown above, as well as against the colon cancer cell line (101/1.5 microL/mL). However, the root extract inhibited SK-LU1 more strongly than did the fruit juice, cucurbitacin B (3), and bryonolic acid (1) (149/169/180/>500 microL/mL, respectively). The root extract inhibited the two lung and three breast cancer cell lines (SKBR3, MDA-MB435, and MCF7) more strongly than the fruit juice. Bryonolic acid (1) inhibited MDA-MB435 somewhat better than the other tested human cancer cell lines. The fruit juice inhibited the colon cancer cell line (Caco-2) more strongly than the root extract. Cucurbitacin (3) inhibited human cancer cell lines, especially Caco-2, much more strongly than bryonolic acid (1). In addition to bryonolic acid (1), bryononic acid (2), cucurbitacin B (3), and dihydrocucurbitacin B (4) also were isolated from the root extract. PMID- 19288407 TI - Effect of donor nerve injury distal to an end-to-side neurorrhaphy model. AB - In end-to-side neurorrhaphy, "noninjury" models of the donor nerve do not seem to offer worthwhile functional outcomes. The role of donor nerve injury distal to the coaptation site remains unclear. End-to-side neurorrhaphy was studied in a rat model in which the proximal stump of the transected musculocutaneous nerve was sutured to the median nerve by end-to-side coaptation. Twenty Sprague-Dawley rats were randomized to four groups of five animals each, in which three different types of donor injury (crush, ligation, or transection injury) distal to the coaptation site were executed; findings were compared with a similar end to-side model without donor nerve injury (control). Behavioral analysis, electrophysiological studies, muscle morphometric studies, and nerve fibers counts showed no significant differences among groups. However, there was a significant difference regarding mean myelin area (P = 0.0362) and mean fiber diameter (P = 0.0159) for the crush injury group as compared with the control group. No significant differences were found among the other groups. These data suggest that donor crush injury distal to the coaptation site may increase the rate of myelin formation in regenerating axons across an end-to-side model; however, at 4 weeks of follow-up, there was no significant behavioral or functional significance in this treatment group. PMID- 19288408 TI - Simultaneous replantation of both lower legs in a child: 23 years later. AB - Function is the single most important determinant in the assessment of the results of extremity replantations. Accordingly, the indications for extremity replantations are based on the prediction of sustained satisfactory functional outcome. There are limited reports in the literature regarding replantations of major segments of lower extremities and, particularly, long-term results of those surgeries. However, this analysis is extremely important in refinement of indications for major limb replantations. In this article, evaluation of a patient 23 years after simultaneous replantation of both lower legs is presented. PMID- 19288409 TI - An overview of skin flap surgery in the mainland China: 20 years' achievements (1981 to 2000). AB - Microsurgical anatomy and clinical applications have been widely and extensively practiced throughout the mainland of China since late 1970s. During the 1980s to 1990s, Chinese surgeons and anatomists developed many new flap donor sites and modifications, most of which were published in Chinese literature. These achievements were not fully realized by the Western surgeons. In this overview, we attempt to give a brief introduction of these contributions made by the Chinese authors in the mainland. Of the new flaps first or independently described by the Chinese, most were in the limbs. These flaps can be classified into three categories. First are free flaps with a main artery trunk, such as the radial forearm flap with radial artery, the medial leg flap with posterior tibial artery, and lateral leg flap with peroneal artery. Second are reverse-flow island flaps based on distal main vascular bundles (e.g., the radial artery and venae comitantes, the ulnar, the posterior tibial, and the peroneal arteries). Third are septocutaneous perforator flaps that avoid sacrifice of the main artery trunk, which include the anterolateral thigh flap, lateral lower-leg flap, dorsoulnar flap, distally adipofascial pedicled radial forearm flap, and so on. PMID- 19288410 TI - Jejunal diverticular bleeding treated with hemoclips at double-balloon enteroscopy. PMID- 19288411 TI - Successful management of a difficult colon stricture with an oblique-view upper scope. PMID- 19288412 TI - Multiple breakage of a covered esophageal stent. PMID- 19288413 TI - Biliary cannulation can be facilitated by pancreatic stent insertion. PMID- 19288414 TI - Direct percutaneous endoscopic jejunostomy using a transgastrostomic endoscope in patients with previous endoscopic gastrostomy. PMID- 19288415 TI - Gastropleural fistula: a complication of esophageal self-expanding metallic stent migration. PMID- 19288416 TI - A very unusual appearance of a rare endoscopic finding: esophageal hematoma. PMID- 19288417 TI - Endocytoscopic imaging and tattooing: a caveat. PMID- 19288418 TI - Contrast-enhanced low mechanical index endoscopic ultrasound (CELMI-EUS). PMID- 19288419 TI - Amyloid tumor of the stomach simulating an obstructing gastric carcinoma: case report and review of the literature. PMID- 19288420 TI - Achalasia associated with multiple esophageal diverticula. PMID- 19288421 TI - A new endoscopic treatment method for a symptomatic duodenal duplication cyst. PMID- 19288422 TI - [Prevalence and incidence of dementia in Germany--a study based on data from the public sick funds in 2002]. AB - So far all calculations of the number of demented people are based on rates from meta-analyses, mean rates of meta-analyses or spatial analyses. This article presents age- and gender-specific prevalence and incidence rates of dementia that are based on a large sample of the German Sick Funds (Stichprobendaten von Versicherten der gesetzlichen Krankenversicherung (GKV)) with 2.3 million people from the year 2002. Prevalence rates increase from 0.8% and 0.6% for 60-64 year old men and women to 30% and 43% for men and women aged 100 or older, respectively. Incidence rates increase from 0.18 and 0.14 cases per 100 person years for 60-64-year old men and women to 9.9 and 10.9 for over 95 year old men and women, respectively. Our results confirm rates from earlier studies on the basis of meta-analyses. Regional differences show for the first time that higher prevalence rates exist for East German women and men above age 85. In 2007 about 1.07 million moderately or severely demented people live in Germany of which about 244 000 are incident cases when we extrapolate our rates to the population of this year. PMID- 19288423 TI - [Complementary system of health care in cooperation with migrant communities- requirements for successful integration in health care]. AB - National and ethnic diversities among and within groups of migrants present great challenges to health care, not only in Germany. Access to regular health care, in particular for relatively small migrant groups and new immigrants is sometimes difficult. In addition, specific cultural aspects and legal implications--such as the residence status--must be taken into account when setting up a complementary system of health care. Methods and arrangements should make use of already existing resources of the health care system as well as proceed to direct cooperation with migrant communities. The public health service with its wide range of technical and organisational efficiency can play a key role here, especially in the municipal sector, both for an effective medical care for those who have difficulties in gaining access to the system, as well as for developing municipal networks, cooperating with a maximum number of medical service providers, organisations, and regarding specific migrant lifestyles. Social and ethic aspects of medical care for people living in precarious conditions ought to be supplemented by practice models which render a flexible and economic organisation of a complementary system of health care, the only way to a sustainable medical care for small migrant groups or not optimally integrated immigrant populations. PMID- 19288424 TI - [Nickel levels in female dermatological patients]. AB - Nickel levels in urine were determined among 163 female dermatological patients aged 18 to 46 years. Data on life-style factors were collected in parallel via a questionnaire. Urinary nickel excretion was in the normal range of the German female population (0.2-46.1 microg Ni/g creatinine). The 95th percentile (3.9 microg Ni/l urine) exceeded the German reference value (3.0 microg Ni/l urine). In the multivariate regression analyses we found a statistically significant increase of ln-transformed nickel levels with increase in age and in women using dietary supplements. The following variables were not associated with Nickel urine levels: suffering from nickel eczema, smoking, drinking stagnated water, eating foods with high nickel contents and using nickel-containing kitchen utensils as, for example, an electric kettle with an open heater coil. We conclude that personal urinary levels should be assessed with simultaneous consideration of habits and life-style factors. A German national survery would be useful. Those patients who experience the exacerbation of their eczema in cases of oral provocation, for example, by a high nickel diet should be aware of potential sources of nickel, such as supplements. PMID- 19288425 TI - [Health economic effects after DRG-implementation--a systematic overview]. AB - BACKGROUND: Since 1 January 2004, inpatient treatment services in German hospitals have been reimbursed using a prospective payment method based on diagnosis-related groups (DRGs) rather than daily rates. The aim of the payment system reform was to decrease the length of inpatient stays and reduce overall healthcare expenditure, the latter of which had increased markedly during previous decades. OBJECTIVE: The primary objective of our study was to analyse and describe the health-economic consequences of implementing a DRG-based system of prospective payment in Germany. METHODS: A systematic search of the literature was performed on MEDLINE. Inclusion criteria were a focus on health economic variables from the German perspective and a publication date after 1 January 2004. The search was supplemented by a manual review of references, as well as internet-based hand search. The main health-economic conclusions were subsequently extracted from all of the included studies. RESULTS: A total of 19 quantitative and qualitative studies were included. There were substantial differences between them in terms of medical focus and hospital characteristics. The most common health-economic variables analysed were revenue generated by patient treatment, and length of inpatient stay. As expected, both variables showed a decreasing trend following the introduction of DRGs. The included studies also investigated the development of case numbers, the proportion of outpatient services provided, the number of diagnoses per case, and the homogeneity of case groups. For these variables, the studies showed a wide range of results. CONCLUSION: Similar to the experience with DRGs in many other countries, the introduction of DRGs in Germany has led to a reduction in the length of inpatient stay and a decrease in hospital revenues. The effects on other health-economic parameters are inconsistent. Additional studies in this area are needed. PMID- 19288426 TI - [The BSD Health Study: a pilot study to examine the comparability of Bavarian blood donors with the Bavarian general population by a comparison with KORA S4]. AB - AIM: The aim of this pilot study was to compare the health status of Bavarian blood donors to the general population in Bavaria (Southern Germany). METHODS: Blood donors in Bavaria were evaluated with respect to socio-demographic characteristics, smoking status and selected diseases. The subset of blood donors from rural areas and aged 35-74 years (n=1 187), was compared to participants in the population-based survey KORA S4 living in rural areas near Augsburg (n=1 870). The comparison was based on logistic regression models adjusted for age and sex. RESULTS: Bavarian blood donors lived significantly more often in communities with less than 20 000 inhabitants (OR=4.73, KI: 3.69-6.13). They were more often born in Germany (OR=2.71, KI: 2.12-3.50) and had less university degrees (OR=0.60, KI: 0.44-0.81). Never smoking is more common in blood donors (OR=1.53, 95% KI: (1.30; 1.80)). The prevalence of asthma, myocardial infarction (HI) and cancer (men only) is significantly lower in blood donors compared to the general population both living in rural areas (OR (Asthma)=0.32, 95% KI: (0.20; 0.52); OR (HI)=0.17, 95% KI: (0.08; 0.34)). The initially significantly lower number of cancer cases in male blood donors disappeared after adjustment for smoking status and community size. No difference was detected for type 2 diabetes. CONCLUSION: With the exception of type 2 dia-betes, the prevalence of the investigated diseases was lower in blood donors than in the general population. Differences can probably be explained by lifestyle factors as there might be a selection of healthier and more health-conscious blood donors. In the literature, a reduced risk of MI is discussed to be related to the frequency of blood donation. However, these questions need further research. PMID- 19288427 TI - [Community analysis serving as a foundation for community-based health promotion programming--results of a citizen survey in Karlshuld, Germany]. AB - BACKGROUND: Analysing a community's needs and requirements is useful for planning a community-based health promotion programme. The literature recommends the analysis of existing population-based data, the discussion with a community's key persons, and standardised surveys among the community members. With regagrd to the latter method, German-language literature is scarce. AIM: A community analysis in the Bavarian rural community Karlshuld (5 000 inhabitants) was intended to provide a starting point for tailored health promotion activities in the community. As a part of the community analysis, a survey among the Karlshuld citizens was intended to inform about (1) health-related knowledge and behaviour of the community members, (2) their utilization and acceptance of existing health related offers and activities, (3) their possibilities for active participation, their state of empowerment and (4) community-related aspects influencing quality of life. METHODS: A standardised telephone survey was performed with a random sample of Karlshuld community members aged 15 years and older. The questions covered health-related living conditions and activities in Karlshuld, quality of life and participation of community members. In addition, data were collected on knowledge, attitude and behaviour with regard to the issues nutrition, physical activity and tobacco consumption. The sample comprised 314 community members (f=68.8%; age 43.9+/-13.8 years). RESULTS: Shortcomings in knowledge on healthy nutrition were evident, especially among male participants. Whereas the general attitude towards a healthy diet was favourable in the majority of respondents, only 45.9%/58.6% stated that they daily consumed vegetables/fruit. One third of the 27.4% smokers among the respondents had intended but failed to quit smoking in the previous year; 80% stated they were willing to quit. The participants named infrastructure, nature and health-related activities as positive aspects in Karlshuld. Only 50% knew where to get information on health-related issues. 42.9% of respondents felt they could influence decisions on health-related issues in the community. There was no correlation between school education and knowledge or participation. DISCUSSION: The survey among Karlshuld community members identified potential approaches to improving the community environment and to tailor health education activities to the special needs of Karlshuld. A follow-up survey in 2008/2009 will be carried out in order to evaluate whether or not the Karlshuld health promotion programme was successful in building on Karlshuld's strengths and influencing its weaknesses. PMID- 19288428 TI - [Are American physicians more satisfied?--results from an International Study of Physicians in University Hospitals]. AB - Understanding the factors that affect physicians' job satisfaction is important not only to physicians themselves, but also to patients, health system managers, and policy makers. Physicians represent the crucial resource in health-care delivery. In order to enhance efficiency and quality in health care, it is indispensable to analyse and consider the motivators of physicians. Physician job satisfaction has significant effects on productivity, the quality of care, and the supply of physicians. The purpose of our study was to assess the associations between work-related monetary and non-monetary factors and physicians' work satisfaction as perceived by similar groups of physicians practicing at academic medical centres in Germany and the U.S.A., two countries that, in spite of differing health-care systems, simultaneously experience problems in maintaining their physician workforce. We used descriptive statistics, factor and correlation analyses to evaluate physicians' responses to a self-administered questionnaire. Our study revealed that overall German physicians were less satisfied than U.S. physicians. With respect to particular work-related predictors of job satisfaction we found that similar factors contributed to job satisfaction in both countries. To improve physicians' satisfaction with working conditions, our results call for the implementation of policies that reduce the time burden on physicians to allow more time for interaction with patients and colleagues, increase monetary incentives, and enhance physicians' participation in the development of care management processes and in managerial decisions that affect patient care. PMID- 19288429 TI - [Scientific inquiry]. PMID- 19288430 TI - [Smoking bans in public places: current epidemiological evidence of cardiovascular health impacts at the population level]. AB - During the past years smoking bans in public places including hospitality venues have been introduced in several countries. Up to now, eight ecological studies on hospital admission rates due to acute myocardial infarction or coronary events after introduction of a smoking ban in the United States, Canada, and Italy have been published. This article reviews these studies and discusses their significance and potential sources of error from an epidemiological point of view. The chronological order of reduction in acute myocardial infarction rates following the smoking ban, the consistency of this association in several countries, and the biological plausibility because of the known acute cardiovascular effects of secondhand smoke suggest a causal association. Thus, if this turns out to be true public smoking bans will be a very effective public health measure. PMID- 19288431 TI - [Patient preferences and willingness to wait for a work-related orthopaedic rehabilitation: a discrete choice experiment]. AB - BACKGROUND: Implementing a stronger patient orientation needs a secured knowledge about wants, preferences and quality standards of the persons concerned. In international health services research discrete choice experiments (DCE) have been used to analyse such complex preferences for some years. The method uses comparisons of two or more services that vary in several attributes to achieve attribute-related decisions to one of the services. From these decisions the relative importance of the attributes considered can be estimated. OBJECTIVE: A DCE was conducted to explore the preferences of rehabilitants concerning the design of orthopaedic rehabilitation. Additionally, the study should enable analyses concerning the theoretical validity of the approach. METHODS: Data collection was realised within a cluster randomised trial examining the efficacy of a work-related orthopaedic rehabilitation. The DCE considered four attributes (offer of a social counselling service, offer of a work-related stress coping training, intensity of therapy, waiting time). For the questionnaire nine choice sets were generated from the potential programmes. RESULTS: The analyses considered data of 223 patients finishing 1796 choice sets. Respondents preferred the offer of a social counselling service (beta=0.20; 95% CI: 0.10 to 0.30), the offer of a work-related stress coping training (beta=0.70; 95% CI: 0.60 to 0.80), rising the intensity from 3 to 5 hours (beta=0.36; 95% CI: 0.21 to 0.50) and reduced waiting time (beta=-0.07; 95% CI: -0.10 to -0.03). Relative importance (RI) was highest for the offer of a stress coping training (RI=45.7%). For such an offer the respondents were willing to wait 10 additional weeks. A model segmented for persons with convenient and less convenient stress coping strategies confirmed higher benefit gains for persons with a lower ability to disassociate oneself from work-related strains (chi(2)=13.27; p<0.001). CONCLUSION: High benefit expectations of employed persons concerning a work related stress coping training within an orthopaedic rehabilitation support that a sufficient acceptance and compliance is expectable when implementing an appropriate intervention. As the realisation of a stronger patient orientation needs knowledge about patient preferences, development of German health services could benefit from conducting DCE. PMID- 19288432 TI - [The quality assurance programme of the statutory health insurance funds in medical rehabilitation: results and further developments]. AB - OBJECTIVE OF THE STUDY: This study reports on the results of the quality assurance programme of the statutory health insurance funds in medical rehabilitation (QS Reha programme) and on the further development of the programme in 2007 to 2008. By October 2008, a total of 240 rehabilitation centres with 283 specialised departments were participating in the QS-Reha programme. METHODOLOGY: To measure structural quality, the level of compliance with "basic criteria" (compiled beforehand with the help of experts) was determined. This was done by means of a questionnaire and subsequent telephone interviews with each institution. To determine patient satisfaction and measure outcome quality, a multicentric study with three measurement periods was carried out. Risk adjustment was conducted to compare clinics. The latest results presented here include the data from about 8,000 patients. RESULTS: On average, the structural quality of the institutions can be described as good to very good (the rate of fulfillment is generally over 90%), but there are clear differences among the clinics. The patients are generally very satisfied with the personnel. The fraction of those not satisfied is about 10-15%. At the end of rehabilitation, medium high to high effects on the central dimensions of outcome can be determined for all the indications observed here. There are also significant deviations from the overall mean for 10-40% of the clinics after risk adjustment. DISCUSSION: The preliminary results of the programme provide a comprehensive view of the quality of medical rehabilitation. Limitations exist concerning evidence based structural quality criteria, methodological problems of patient-reported outcomes and the occurrence of non-responders and dropouts. The reworking of the QS Reha programme pursued the goal of improving the cost-benefit ratio of compiling data without impairing the methodological soundness of the programme. The new concept is scheduled to be implemented routinely beginning in 2009. PMID- 19288433 TI - [Never forget this in making your drawings and equations! A conversation with Albert Einstein on learning, teaching and the secrets of the world]. AB - Albert Einstein, the genius--this aspect often has been noted. A neglected aspect is Einstein's role as student and teacher. For this reason, Einstein's notes have been looked at once again. The selected original quotes are composed into the format of a fictive dialogue. The original context and coherence of his comments have thereby been respected carefully. PMID- 19288434 TI - Parasitology and necropsy of fish. AB - Parasitic diseases are common in fish. Diagnosis can be made through gill biopsy, skin cytology, fecal examination, or necropsy. Common parasites include protozoa, helminths, and crustaceans. Determining the cause of death in a fish is important for maintaining the health of other fish in the same environment. Due to rapid autolysis, fish necropsies should be performed promptly after death. Samples should be preserved in 10% neutral buffered formalin. Squash preparations, tissue imprints, microbiology, and virology are also useful in obtaining a diagnosis. PMID- 19288435 TI - Anesthesia, diagnostic imaging, and surgery of fish. AB - Anesthesia, diagnostic imaging, and surgery of fish have become routine parts of aquatic animal medicine. Anesthesia may be required for simple clinical procedures, diagnostic testing, or more involved surgery. Diagnostic modalities, including radiology, ultrasonography, and endoscopy, can be readily applied to fish and may provide valuable information. Despite some unique challenges, surgery can be performed in fish using basic surgical skills and principles and should be considered as a valid treatment option. PMID- 19288436 TI - Laparoscopic-assisted and laparoscopic prophylactic gastropexy: indications and techniques. PMID- 19288437 TI - Behavior assessment: completing the examination. AB - The first two articles in this series on assessing behavior patients explored the use of history forms and owner interviews to gather information about history, environment, other problems, and owner commitment to treatment. This article addresses direct observation and evaluation of the patient, including interaction with the owner, veterinary staff, and other pets. PMID- 19288438 TI - Oral examination of cats and dogs. PMID- 19288439 TI - Obstructive lesions and traumatic injuries of the canine and feline tracheas. AB - Tracheal obstruction and tracheal injury can be life-threatening conditions in dogs and cats. Early identification of associated clinical signs and the use of appropriate diagnostic and therapeutic techniques are important to patient survival. Common causes of tracheal obstruction reported in the veterinary literature include tracheal foreign bodies and tracheal tumors. Tracheal injury has been associated with endotracheal intubation and external trauma. Radiography and tracheoscopy are effective diagnostic modalities, and tracheal repair or resection and anastomosis are the most common treatments. PMID- 19288440 TI - Modeling fluid flow through irregular scaffolds for perfusion bioreactors. AB - Direct perfusion of 3D tissue engineered constructs is known to enhance osteogenesis, which can be partly attributed to enhanced nutrient and waste transport. In addition flow mediated shear stresses are known to upregulate osteogenic differentiation and mineralization. A quantification of the hydrodynamic environment is therefore crucial to interpret and compare results of in vitro bioreactor experiments. This study aims to deal with the pitfalls of numerical model preparation of highly complex 3D bone scaffold structures and aims to provide more accurate wall shear stress (WSS) estimates. microCT imaging techniques were used to reconstruct the geometry of both a titanium (Ti) and a hydroxyapatite scaffold, starting from 430 images with a resolution of 8 microm. To tackle the tradeoff between model size and mesh resolution we selected two concentric regions of interest (cubes with a volume of 1 and 3.375 mm(3), respectively) for both scaffolds. A flow guidance in front of the real inlet surface of the scaffold was designed to mimic realistic inlet conditions. With a flow rate of 0.04 mL/min perfused through a 5 mm diameter scaffold at an inlet velocity of 33.95 microm/s we obtained average WSSs of 1.10 and 1.46 mPa for the 1 mm(3) and the 3.375 mm(3) model of the hydroxyapatite scaffold compared to 1.40 and 1.95 mPa for the 1 mm(3) model and the 3.375 mm(3) model of the Ti scaffold, showing the important influence of the scaffold micro-architecture heterogeneity and the proximity of boundaries. To assess that influence we selected cubic portions, of which the WSS data were analyzed, with the same size and the same location within both 1 and 3.375 mm(3) cubic models. Varying the size of the inner portions simultaneously in both model selections gives a quantification of the sensitivity to boundary neighborhood. This methodology allows to get more insight in the complex concept of tissue engineering and will likely help to understand and eventually improve the fluid-mechanical aspects. PMID- 19288441 TI - Localized expression profiles of rpoS in Escherichia coli biofilms. AB - Although importance of the rpoS gene on biofilm formation by Escherichia coli has been suggested, there has not been any report showing where the rpoS is expressed during biofilm formation process. Since physiological state of the cells in the biofilms is considerably heterogeneous, the expression of the rpoS gene must be heterogeneous. In this study, in situ spatial expression of the rpoS gene during biofilm formation was investigated with an rpoS-gfp transcriptional fusion mutant strain. A ribosomal binding site and a gene encoding a green fluorescent protein were introduced into the downstream of the rpoS gene, which enabled us to observe the in situ spatial expression of the rpoS gene during biofilm formation processes without any disturbance of the rpoS expression. In the early stages of the biofilm formation process, the rpoS gene was expressed in the most of the cells. On the other hand, the rpoS expression was observed only at the outside of the biofilms during the late stages of the biofilm formation process. The in situ spatial expression of the rpoS gene in the biofilm was verified by quantifying the expression levels of the rpoS at the outside and the inside of the biofilms with the real time RT-PCR. In addition, global gene expression analysis was performed with DNA microarray to investigate physiological difference between the outside and the inside of the biofilms. This heterogeneous rpoS expression profile suggested that the cells at the outside of the biofilm need to express the rpoS to shift the physiological state to the stationary growth mode such as induction of various stress responses and suppression of the motility. PMID- 19288442 TI - A novel approach for enhancing the catalytic efficiency of a protease at low temperature: reduction in substrate inhibition by chemical modification. AB - The alkaline protease, savinase was chemically modified to enhance the productivity of the enzyme at low temperatures on a complex polymeric protein (azocasein) substrate. At 5 and 15 degrees C, savinase modified with ficol or dextran hydrolyzed fivefold more azocasein than the unmodified savinase. Kinetic studies showed that the catalytic improvements are associated with changes in uncompetitive substrate inhibition with K(i) values of modified savinases sixfold higher than the unmodified savinase. Modeling of small-angle scattering data indicates that two substrate molecules bind on opposing sides of the enzyme. The combined kinetic and structural data indicate that the polysaccharide modifier sterically blocks the allosteric site and reduces substrate inhibition. In contrast to the properties of cold-active enzymes that generally manifest as low activation enthalpy and high flexibility, this study shows that increased activity and productivity at low temperature can be achieved by reducing uncompetitive substrate inhibition, and that this can be achieved using chemical modification with an enzyme in a commercial enzyme-formulation. PMID- 19288443 TI - Thermodynamic analysis of biodegradation pathways. AB - Microorganisms provide a wealth of biodegradative potential in the reduction and elimination of xenobiotic compounds in the environment. One useful metric to evaluate potential biodegradation pathways is thermodynamic feasibility. However, experimental data for the thermodynamic properties of xenobiotics is scarce. The present work uses a group contribution method to study the thermodynamic properties of the University of Minnesota Biocatalysis/Biodegradation Database. The Gibbs free energies of formation and reaction are estimated for 914 compounds (81%) and 902 reactions (75%), respectively, in the database. The reactions are classified based on the minimum and maximum Gibbs free energy values, which accounts for uncertainty in the free energy estimates and a feasible concentration range relevant to biodegradation. Using the free energy estimates, the cumulative free energy change of 89 biodegradation pathways (51%) in the database could be estimated. A comparison of the likelihood of the biotransformation rules in the Pathway Prediction System and their thermodynamic feasibility was then carried out. This analysis revealed that when evaluating the feasibility of biodegradation pathways, it is important to consider the thermodynamic topology of the reactions in the context of the complete pathway. Group contribution is shown to be a viable tool for estimating, a priori, the thermodynamic feasibility and the relative likelihood of alternative biodegradation reactions. This work offers a useful tool to a broad range of researchers interested in estimating the feasibility of the reactions in existing or novel biodegradation pathways. PMID- 19288444 TI - Faster clearance of omeprazole in mutant Nagase analbuminemic rats: possible roles of increased protein expression of hepatic CYP1A2 and lower plasma protein binding. AB - It is well known that there are various changes in the expression of hepatic and intestinal CYPs in mutant Nagase analbuminemic rats (NARs). It has been reported that the protein expression of hepatic CYP1A2 was increased, whereas that of hepatic CYP3A1 was not altered, and it was also found that the protein expression of the intestinal CYP1A subfamily significantly increased in NARs from our other study. In addition, in this study additional information about CYP changes in NARs was obtained; the protein expression of the hepatic CYP2D subfamily was not altered, but that of the intestinal CYP3A subfamily increased in NARs. Because omeprazole is metabolized via hepatic CYP1A1/2, 2D1, 3A1/2 in rats, it could be expected that the pharmacokinetics of omeprazole would be altered in NARs. After intravenous administration of omeprazole to NARs, the Cl(nr) was significantly faster than in the controls (110 versus 46.6 ml/min/kg), and this could be due to an increase in hepatic metabolism caused by a greater hepatic CYP1A2 level in addition to greater free fractions of the drug in NARs. After oral administration of omeprazole to NARs, the AUC was also significantly smaller (80.1% decrease) and F was decreased in NARs. This could be primarily due to increased hepatic and intestinal metabolism caused by greater hepatic CYP1A2 and intestinal CYP1A and 3A levels. In particular, the smaller F could mainly result from greater hepatic and intestinal first-pass effect in NARs than in the controls. PMID- 19288445 TI - Novel endotoxin assay by laser light-scattering particle-counting method. AB - Exposure of Limulus amoebocyte lysate to endotoxin under stirring produced light reflective particles that appeared to be coagulin polymers. A laser light scattering particle counter, the PA-200, detected these particles sensitively. The PA-200 detected endotoxin at a concentration as low as 0.00015 EU/ml in 71 min, whereas the minimum endotoxin concentration measured by a turbidimeter, ET 2000, was 0.0005 EU/ml in 138 min. Moreover, PA-200 was much less affected by the presence of colored substances and refractive materials than was ET-2000. We propose that the high sensitivity, speed, and high interference tolerance of the laser light-scattering particle-counting method make it more useful than the widely used turbidimetric method for quantitative endotoxin assay. PMID- 19288446 TI - Associations of IgG N-linked oligosaccharide chains and proteases in sera of prostate cancer patients with and without alpha2-macroglobulin deficiency. AB - We previously reported on a number of cases of metastatic prostate cancer (PCa) in which serum alpha2-macroglobulin (alpha2M) levels were markedly decreased to less than 20 mg/dl (alpha2M deficiency). All PCa patients with alpha2M deficiency had multiple bone metastases. Proteases in ten PCa patients with and without alpha2M deficiency were studied and compared against ten healthy controls in order to elucidate the relationships between changes in sugar chain structure and neoplasia. We assessed the relationship between ratios of Fr4 to Fr1 and Fr2 (Fr4/Fr1+Fr2 ratios) of oligosaccharide chains, and ratios of free prostate specific antigen (PSA) to total PSA (F/T ratios), and serum levels of matrix metalloproteinase-2 (MMP-2) in PCa progression. Measurement of serum alpha2M concentration was performed by laser nephelometry. Serum PSA and MMP-2 levels were determined by enzyme immunoassay and free PSA by radioimmunoassay. N-linked oligosaccharides of human serum immunoglobulin G were analyzed using fluorophore associated carbohydrate electrophoresis. In those PCa patients with alpha2M deficiency: (a) serum alpha2M and F/T ratios were lower (P<0.05) and (b) Fr4/Fr1+Fr2 ratios and serum MMP-2 levels were higher when compared with those PCa patients without alpha2M deficiency. There was a significant correlation between Fr4/Fr1+Fr2 ratios and F/T ratios or serum MMP-2 levels in PCa with alpha2M deficiency (P<0.05). Therefore, these markers may serve as an auxiliary serum tumor marker for monitoring of the bone metastases or progression of disease in PCa. PMID- 19288447 TI - Serum prolidase activity and oxidative status in patients with bronchial asthma. AB - Asthma is a disease where there is an accumulation of collagen in the reticular basal membrane of the airway leading to chronic inflammation. The enzyme prolidase plays an important role in the breakdown of collagen and the breakdown of intracellular protein especially in the final stage when peptides and dipeptides contain a high level of proline. To evaluate the relationship between prolidase activity and oxidative status in asthma patients. Comparison was made between 42 patients diagnosed with bronchial asthma and 32 healthy children of similar age and gender. Serum prolidase activity was measured spectrophotometrically. Oxidative status was determined using total antioxidant capacity (TAC) and total oxidant status (TOS) measurement. The prolidase activity of the asthma patient group was statistically significant compared with the control group (P< or =0.001). TAC and TOS levels in the asthma patient group were higher than the control group (P< or =0.001, P< or =0.002, respectively). No correlation was found between the prolidase and oxidative levels of the two groups. A positive correlation was determined between the prolidase activity and TAC in the asthma patient group (P< or =0.001, r=0.501). The prolidase enzyme activity, which plays a role in the collagen turnover, was low in the asthma patients; therefore, their collagen metabolism had undergone a change and this indicates that there may be an effect on the accumulation of collagen in the reticular basal membrane. Moreover, the high level of TOS indicates that these patients were exposed to severe oxidative stress with an increased TAC response. PMID- 19288448 TI - Detection of Paracoccidioides brasiliensis gp43 gene in sputa by loop-mediated isothermal amplification method. AB - The fungus Paracoccidioides brasiliensis is the pathogen of paracoccidioidomycosis (PCM), a systemic mycosis prevalent in Latin America. The loop-mediated isothermal amplification method (LAMP) was used in this study to detect the presence of P. brasiliensis in sputa samples from patients with chronic PCM, suspected PCM, and a negative control. The target P. brasiliensis gp43 gene was amplified in less than 4 hr in 11 of 18 sputa samples tested. The LAMP method had the advantage of speed and simplicity compared with the classic diagnostic methods such as the histopathological test or biological material culture and did not require sophisticated technical apparatus. It would be an important aid in cases where immediate treatment would mean patient survival, especially in immune-suppressed patients. PMID- 19288449 TI - Interleukin-18 gene 105A/C genetic polymorphism is associated with the susceptibility of Kawasaki disease. AB - Interleukin-18 (IL-18)-656T/G, -607A/C, and -137C/G promoter polymorphisms had been reported associated with Kawasaki disease (KD). An IL-18 genetic A/C polymorphism at coding position 105 (rs549908) has been linked with asthma, rheumatoid, and systemic lupus erythematosus. We tested a hypothesis that the IL 18 105A/C genetic polymorphism confers KD susceptibility. Study participants were Taiwanese KD patients and a healthy control group. Our data indicated that the frequency of C allele was significantly higher in the patient group (13.9%) than in the control group (2.7%; P<0.0001, odds ratio [OR]=5.93; 95% confidence interval [CI]=2.57-13.73). Therefore, persons with the C allele may have higher risk of developing KD. In addition, compared with the haplotype frequencies between case and control groups, the KD patients with TACC haplotype appeared to be a significant "at-risk" haplotype compared with other haplotypes (OR: 4.62, 95% CI: 1.71-12.43; P=0.001). KD patient with the TAGA haplotype appeared to be a significant "protective" haplotype compared with other haplotypes (OR: 0.51, 95% CI:0.29-0.89; P=0.017). Our results suggest that 105A/C polymorphism and the haplotypes in IL-18 gene are associated with the risk of KD in Taiwanese population. PMID- 19288450 TI - Transferrin D protein variants in the diagnosis of congenital disorders of glycosylation (CDG). AB - Congenital disorders of glycosylation are a rapidly growing group of inherited (neuro)metabolic disorders characterized by defects in glycosylation of proteins and lipids. This study discusses an analytical problem in the differentiation between hypoglycosylation and transferrin (Tf) protein variants. Analysis of serum Tf by isoelectric focusing is used as a common method suitable for screening 19 out of a total of 22 types of glycosylation defects identified so far. In three members of a family, several indicators showed evidence of a Tf protein variant, however, routine neuraminidase-based demonstration failed to confirm this result. On the assumption that we should be able to exclude Tf protein variants at the screening-level of the diagnostic algorithm, our concern is a possible cause of our failure to confirm some of the Tf D variants (in contrast to the other C, B and D allelic combinations that are commonly well identified). Several explanations are discussed. PMID- 19288451 TI - Diagnostic assay of chromium (VI) in the ex vivo fluid of the urine of a smoker using a fluorine-doped handmade sensor. AB - A voltammetric diagnosis of a chromium (VI) ion was investigated using a fluorine doped graphite pencil electrode. Square wave (SW) stripping working conditions were attained at a high range of 0.051-0.45 mg L(-1) and a microrange of 0.05-0.4 microg L(-1) in a 0.1 M NH(4)H(2)PO(4) electrolyte solution, at a relative standard deviation of 1.68% (RSD, n=15), using 10.0 microg L(-1) Cr(VI). A fast experimental time was used only for the 120 sec SW accumulation time. An analytical detection-limit (DL) of 0.008 microg L(-1) was attained. DL appeared to be more sensitive than common voltammetric and spectrophotometric assays. The developed sensor was applied to tap water and the urine of a smoker. It was found that the methods can be applicable for in vivo fluid or medicinal diagnosis. PMID- 19288452 TI - Evaluation of metalloproteinases 2 and 9 and their inhibitors in physiologic and pre-eclamptic pregnancy. AB - Matrix metalloproteinases (MMPs) are a family of zinc and calcium-dependent endopeptidases involved in remodeling and physiological homeostasis of extracellular matrix (ECM). The metalloproteinases activity is predominantly modulated by specific tissue inhibitors of matrix metalloproteinases (TIMPs). The balance between MMPs and TIMPs is likely to play an important role in remodeling uterine arteries in pregnancy, and it may represent means by which vasodilatation is maintained in later pregnancy. Moreover, increased levels of MMPs and in particular MMP-2 play a role in the vascular alterations induced by hypertension. The aim of this study was the evaluation of MMP-2 and -9, along with their inhibitors TIMP-1 and -2, in pre-eclamptic women compared with normotensive pregnancy and non-pregnant women. Fourteen pre-eclamptic women were compared with 37 normotensive women in different gestational age and 21 non-pregnant women. Multiplexed sandwich enzyme-linked immunosorbent assay was used to measure MMPs and TIMPs simultaneously. MMP-2 levels were significantly higher in pre-eclamptic women vs. both non-pregnant and physiologic pregnant women. MMP-9 concentrations were significantly higher in physiologic pregnant vs. non-pregnant women. The serum levels of TIMP-1 were significantly higher in pre-eclamptic vs. both non pregnant and physiologic pregnant women. TIMP-2 values were higher in physiologic pregnant women and pre-eclamptic women vs. non-pregnant women. A positive correlation between MMP-9 values and gestational age was observed in normal pregnant women. Results of the present investigation confirm that MMP-2 and TIMP 1 values are significantly higher in preeclampsia. We confirm that the modification of the fine balance between MMPs and their inhibitors plays a greater role in the structural and functional vascular changes of women with complicated pregnancies. PMID- 19288453 TI - Effect of sample storage on stability of salivary glutathione, lipid peroxidation levels, and tissue factor activity. AB - Saliva samples are often required to be stored for longer periods of time either because of the project protocol or because of lack of funding for analysis. The effects of 6 months storage (fresh, 30, 60, 90 120, 150, and 180 d) on the stability of salivary reduced glutathione (GSH), lipid peroxidation (LPO) and 90 days of storage (fresh, 15, 30, 60, and 90 d) on the stability of salivary tissue factor (TF) activity and the stability of saliva imprint samples at -20 degrees C were evaluated in this study. Salivary GSH, malondialdehyde (MDA) levels as an index of LPO, and TF activities were determined using the methods of Beutler, Yagi, and Quick, respectively. Saliva imprint samples were stained with Giemsa and microscopically examined. Salivary GSH levels and TF activities decreased, whereas MDA levels increased significantly after 6 months of storage at -20 degrees C. Leucocyte, epithelium and bacterium cell counts did not significantly change at the end of 90 d of storage. Saliva samples may be stored up to 1 month at -20 degrees C for LPO assay. For cytological examinations, saliva samples may be stored for 90 d at -20 degrees C. Further studies are needed to determine the stability of salivary GSH, and salivary TF activity stored less than 30 days at 20 degrees C. On the other hand, if saliva samples are required to be stored, to avoid the changes because of different storage periods, we recommend that they must be stored under the same circumstances and in the same time period. PMID- 19288454 TI - Lipid peroxidation and antioxidant enzyme activities in infertile men: correlation with semen parameter. AB - The aim of this study was to investigate the oxidative stress status and antioxidant enzyme activities in infertile men's semen and to determine their relationship with spermatozoa characteristics. Four groups of infertile men, normozoospermic (n=9), azoospermic (n=13), oligoasthenozoospermic (n=38), and asthenozoospermic (n=42), were tested for malonaldialdehyde (MDA) concentration and catalase (CAT) and superoxide dismutase (SOD) activities in semen using spectrophotometric methods. We found that MDA levels in semen and SOD activity in seminal plasma (SP) were significantly higher in oligoasthenozoospermic and asthenozoospermic groups compared with normozoospermic group. In contrast, the mean values of CAT activity were not significantly different in all groups. We found positive correlations between semen MDA concentration and SOD activity (P<0.01). MDA levels in sperm pellet and in SP were also positively correlated with mobility grade b (P<0.01), acrosome anomalies (P<0.01), and residual cytoplasmic droplets (P<0.05). In contrast, SOD activity in SP was negatively correlated with sperm concentration and normal morphology (P<0.05). Similarly, CAT activity in SP was negatively correlated with sperm and leukocyte concentrations (P<0.05). In conclusion, our results suggest that the evaluation of oxidative status and antioxidant defenses may be taken as an important tool for diagnosis and treatment of male infertility. PMID- 19288455 TI - Serum lipid profile, oxidative status, and paraoxonase 1 activity in hyperemesis gravidarum. AB - The aim of this study was to investigate lipid profile, paraoxonase 1 (PON1) activity, and oxidative stress status in the serum of hyperemesis gravidarum (HG) patients. Thirty-six HG cases and 36 normal pregnants were included in the study. Serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apoproteins A1 (apo A1) and B (apo B), malondialdehyde (MDA), and total antioxidant activity (TAO) values and PON1 and arylesterase activities were determined. Although serum TC, TG, LDL-C, and apo B levels were not different among; the groups (P>0.05), HDL-C (P=0.01) and apo A1 (P=0.007) levels were lower in HG patients than in normal pregnants. HG group had significantly lower serum PON1 (P=0.03) and arylesterase activities (P=0.03) compared with the control group. Additionally, mean TAO values were lower (P=0.01) and MDA levels were higher (P=0.02) in HG group than in the healthy pregnants. A significant negative correlation between PON1 and MDA was found in HG group (r=-0.33, P<0.05). The findings of this study have revealed that HG may be one of the conditions in which oxidant and antioxidant balance is impaired. PMID- 19288456 TI - Detection of C1236T, G2677T/A, and C3435T polymorphism of MDR1 by amplification refractory mutation system PCR. AB - C1236T, G2677T/A, and C3435T polymorphism of the multidrug resistance (MDR1) gene have substantial impact on expression or activity of P-glycoprotein (P-gp). We developed new methods based on amplification refractory mutation system (ARMS) to detect these polymorphisms. Tetra-primers amplification in a single tube was established to detect C1236T and C3435T polymorphism. For G2677T/A polymorphism, a two-step allele-specific amplification method was used. MDR1 genotypes of 177 Chinese subjects were determined by the methods we established. The methods we established were verified with gene sequencing. Gene frequencies of 1236C and 1236T were 37.8 and 62.2%, respectively; gene frequencies of 2677G, 2677T and 2677A were 44.1, 38.4 and 17.5%, respectively; the gene frequencies of 3435C and 3435T were 65.0 and 35.0%, respectively. The results were similar with other studies on Oriental subjects. The methods we established are simple, accurate, and economical, and can provide reliable approaches for determining MDR1 polymorphism. PMID- 19288457 TI - Performance of first-trimester combined screening for trisomy 13 and 18 with the double test taken at a gestational age of 8 + 0 to 13 + 6. AB - OBJECTIVE: To evaluate the performance of the combined test for first-trimester screening for trisomy 18 and 13, when the double test is scheduled weeks before the nuchal translucency scanning. METHODS: The study included all 40 cases of trisomy 18 and 13 from April 2004 to October 2008 in a screening programme, where the double test was measured in gestational weeks 8 + 0 to 13 + 6 and the nuchal translucency in weeks 11 + 2 to 13 + 6. RESULTS: Twenty-eight among the 40 cases had complete information on all variables in the first-trimester screening test. Among 19 cases having the double test taken before 10 + 0 weeks, 10 cases were detected (detection rate (DR) = 53%) and among 9 cases having the double test taken after 10 + 0 weeks, 6 cases were detected (DR 67%). There was no significant difference in the DRs (p = 0.48). A total of 29 cases were detected at the first-trimester screening, resulting in an overall DR for trisomy 18 and 13 at 73%. CONCLUSION: This study showed no significant differences in the trisomy 18/13 DRs when grouped according to having the double test taken before or after 10 + 0 weeks. The DR was 73% at the first-trimester screening. PMID- 19288458 TI - Integrating Fusarium oxysporum f. sp. strigae into cereal cropping systems in Africa. AB - BACKGROUND: Striga hermonthica (Del.) Benth. (witchweed) poses the greatest biological constraint to food production in sub-Saharan Africa (SSA). Control options for Striga are currently largely ineffective or unavailable to farmers, and other management possibilities are urgently needed. Biological control obviates some of the problems of several of the other techniques and provides a management option that is durable and environmentally responsive. The efficacy of S. hermonthica control using different formulations of three isolates of Fusarium oxysporum Schlecht. emend. Synder & Hans f. sp. strigae was tested on Striga resistant and Striga-susceptible varieties of sorghum and maize under African field conditions for the first time. RESULTS: Isolates PSM197 and Foxy 2 were effective in witchweed repression, especially when applied as pesta granules. Isolate M12-4A was less effective under the field conditions investigated. Application of the fungi was generally more beneficial in maize than in sorghum for the varieties tested. Application of the biocontrol agent caused significant decreases in the number of flowering Striga plants, and hence deposition of seeds with impact of enhancing future crop yield. CONCLUSIONS: Synergistic effects between the Striga-resistant maize line and Fusarium oxysporum f. sp strigae led to over 90% reduction in Striga emergence. These results will further encourage the distribution of the isolates tested or selection of country-specific relatives as viable and environmentally safe biocontrol agents to be used against Striga. Pesta was the most effective formulation, while seed coating may be more cost effective. PMID- 19288459 TI - In vitro and in vivo evaluation of small interference RNA-mediated gynaecophoral canal protein silencing in Schistosoma japonicum. AB - BACKGROUND: Schistosomiasis causes liver and intestinal damage and can be very debilitating. The pairing of a male worm with a female worm residing in the gynaecophoral canal of male plays a critical role in the development of female parasite. Because the male specific gynaecophoral canal protein of Schistosoma japonicum (SjGCP) is found in significant quantities in the adult female worm after pairing, it could play an important role in parasite pairing. METHODS: In the present study, three small interfering (si)RNA duplexes targeting the SjGCP gene were designed, synthesized and the silencing effects were evaluated in vitro as well as in mice infected with S. japonicum in vivo. RESULTS: In vitro studies using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time RT-PCR revealed the reduction of SjGCP at the transcript level. Similarly, western blotting and immunofluorescence studies showed its reduction at the protein level after treatment of parasites with siRNAs. At a concentration of 200 nM, two siRNAs totally abolished the parasite pairing. To evaluate such a pairing inhibitory effect in vivo, mice infected with S. japonicum were treated with siRNA and both parasite pairing and burden were evaluated. In vivo tests confirmed the in vitro silencing effect of SjGCP siRNA and revealed that the systemic delivery of siRNA significantly inhibited early parasite pairing and the associated burden. CONCLUSIONS: Our preliminary results demonstrated that the SjGCP plays an important role in pairing and subsequent development in S. japonicum, and its silencing might have potential as a therapeutic approach for controlling schistosomiasis. PMID- 19288460 TI - Peptide purification by affinity chromatography based on alpha-ketoacyl group chemistry. AB - Significant advances have been achieved in the fields of peptide/protein synthesis, permitting the preparation of large, complex molecules. Shortcomings, however, continue to exist in the area of peptide purification. This paper details some studies we undertook to develop a new strategy for peptide purification based on a reactivity of alpha-ketoacyl groups in peptides. The alpha-ketoacyl peptide was generated from N(epsilon)-acyl-lysyl-peptide in the solid phase via a transamination reaction using glyoxylic acid and nickel(II) ion. Cleavage of the alpha-ketoacyl group with o-phenylenediamine gave the target peptide in an acceptable yield and purity. We first carried out a careful step-by step optimization of the purification conditions using a model peptide. The strategy was then used in the purification of a transmembrane peptide that could not be effectively purified using a conventional RP-HPLC system due to the strong hydrophobicity of the peptide and its high tendency to aggregate. PMID- 19288461 TI - Synthesis of the C-terminal penta- peptide of the peptaibol culicinins. AB - The synthesis of the C-terminal pentapeptide of culicinins has been achieved using [4 + 1] protocol and reduction-coupling strategy. PMID- 19288462 TI - Functional and effective connectivity of visuomotor control systems demonstrated using generalized partial least squares and structural equation modeling. AB - Tasks employing parametric variation in movement rate are associated with predictable modulations in neural activity and provide a convenient context for developing new techniques for system identification. Using a multistage approach, we explored the functional and effective connectivity of a visuomotor control system by combining generalized partial least squares (gPLS) with subsequent structural equation modeling (SEM) to reveal the relationships between neural activity and finger movement rate in an experiment involving visually paced left or right thumb flexion. The gPLS in the first analysis stage automatically identified spatially distributed sets of BOLD-contrast signal changes using linear combinations of sigmoidal basis functions parameterized by kinematic variables. The gPLS provided superior sensitivity in detecting task-related functional activity patterns via a step-wise comparison with both classical linear modeling and behavior correlation analysis. These activity patterns were used in the second analysis stage, which employed SEM to characterize the areal regional interactions. The hybrid gPLS/SEM procedure allowed modeling of complex regional interactions in a network including primary motor cortex, premotor areas, cerebellum, thalamus, and basal ganglia, with differential activity modulations with respect to rate observed in the corticocerebellar and corticostriate subsystems. This effective connectivity analysis of visuomotor control circuits showed that both the left and right corticocerebellar and corticostriate circuits exhibited movement rate-related modulation. The identification of the functional connectivity among regions participating particular classes of behavior using gPLS, followed by the estimation of the effective connectivity using SEM is an efficient means to characterize the neural interactions underlying variations in sensorimotor behavior. PMID- 19288463 TI - Magnetoencephalographic gamma power reduction in patients with schizophrenia during resting condition. AB - OBJECTIVE: The "default network" represents a baseline condition of brain function and is of interest in schizophrenia research because its component brain regions are believed to be aberrant in the disorder. We hypothesized that magnetoencephalographic (MEG) source localization analysis would reveal abnormal resting activity within particular frequency bands in schizophrenia. EXPERIMENTAL DESIGN: Eyes-closed resting state MEG signals were collected for two comparison groups. Patients with schizophrenia (N = 38) were age-gender matched with healthy control subjects (N = 38), and with a group of unmedicated unaffected siblings of patients with schizophrenia (N = 38). To localize 3D-brain regional differences, synthetic aperture magnetometry was calculated across established frequency bands as follows: delta (0.9-4 Hz), theta (4-8 Hz), alpha (8-14 Hz), beta (14-30 Hz), gamma (30-80 Hz), and super-gamma (80-150 Hz). PRINCIPLE OBSERVATIONS: Patients with schizophrenia showed significantly reduced activation in the gamma frequency band in the posterior region of the medial parietal cortex. As a group, unaffected siblings of schizophrenia patients also showed significantly reduced activation in the gamma bandwidth across similar brain regions. Moreover, using the significant region for the patients and examining the gamma band power gave an odds ratio of 6:1 for reductions of two standard deviations from the mean. This suggests that the measure might be the basis of an intermediate phenotype. CONCLUSIONS: MEG resting state analysis adds to the evidence that schizophrenic patients experience this condition very differently than healthy controls. Whether this baseline difference relates to network abnormalities remains to be seen. PMID- 19288464 TI - Tuning of the visual word processing system: distinct developmental ERP and fMRI effects. AB - Visual tuning for words vs. symbol strings yields complementary increases of fast occipito-temporal activity (N1 or N170) in the event-related potential (ERP), and posterior-anterior gradients of increasing word-specific activity with functional magnetic resonance imaging (fMRI) in the visual word form system (VWFS). However, correlation of these coarse ERP and fMRI tuning responses seems limited to the most anterior part of the VWFS in adult and adolescent readers (Brem et al. [ 2006]: Neuroimage 29:822-837). We thus focused on fMRI tuning gradients of young readers with their more pronounced ERP print tuning, and compared developmental aspects of ERP and fMRI response tuning in the VWFS. Children (10.3 y, n = 19), adolescents (16.2 y, n = 13) and adults (25.2 y, n = 18) were tested with the same implicit reading paradigm using counterbalanced ERP and fMRI imaging. The word-specific occipito-temporal N1 specialization, its corresponding source activity, as well as the integrated source activity (0-700 ms) were most prominent in children and showed a marked decrease with age. The posterior anterior fMRI gradient of word-specific activity instead which was fully established in children did not develop further, but exhibited a dependence on reading skills independent of age. To conclude, prominent developmental dissociation of the ERP and fMRI tuning patterns emerged despite convergent VWFS localization. The ERP response may selectively reflect fast visual aspects of print specialization, which become less important with age, while the fMRI response seems dominated by integrated task- and reading-related activations in the same regions. PMID- 19288465 TI - Functional abnormalities in the dyslexic brain: a quantitative meta-analysis of neuroimaging studies. AB - This study used foci from 17 original studies on functional abnormalities in the dyslexic brain to identify brain regions with consistent under- or overactivation. Studies were included when reading or reading-related tasks were performed on visually presented stimuli and when results reported coordinates for group differences. Activation likelihood estimation (ALE) was used for quantification. Maxima of underactivation were found in inferior parietal, superior temporal, middle and inferior temporal, and fusiform regions of the left hemisphere. With respect to left frontal abnormalities, we found underactivation in the inferior frontal gyrus to be accompanied by overactivation in the primary motor cortex and the anterior insula. Tentative functional interpretations of the activation abnormalities are provided. PMID- 19288466 TI - Functional interactions guiding adaptive processing of behavioral significance. AB - The ability to quickly decide on the nature of unexpected environmental changes is vital for adaptive behavior. Converging evidence suggests that the orbitofrontal cortex plays an important role in the rapid assignment of motivational significance and goal relevance to environmental stimuli. However, its putative role as a central part of a network involved in the prioritization of attentional selection, particulary when significant environmental changes occur unexpectedly or outside of attentional focus, remains to be established. Therefore, we used functional magnetic resonance imaging with a subsequent psychophysiological interaction analysis to reveal the functional connectivity of the right posterior orbitofrontal cortex (pOFC) in a context, in which subjects had to adjust goal-directed behavior to behaviorally relevant events presented outside of the current attentional focus. As expected, an increased functional interaction between pOFC and regions involved in the modulation of selective attention (pulvinar nucleus and inferior parietal lobule) and processing of "bottom-up" salience (substantia nigra) could be observed when unattended, but significant changes were relevant for behavior. Moreover, a positive correlation between level of accuracy and an increased functional connectivity between pOFC and extrastriate cortex suggested that a motivationally-triggered signal from pOFC may have increased visual processing of the relevant but currently unattended stimulus attribute. These data provide evidence that the interplay between the pOFC and these regions underlies a mechanism by which organisms rapidly achieve voluntary control of attentional resources to deal with behaviorally significant changes that occur outside of current attentional focus. PMID- 19288467 TI - Efficacy of systemic morpholino exon-skipping in Duchenne dystrophy dogs. AB - OBJECTIVE: Duchenne muscular dystrophy (DMD) is caused by the inability to produce dystrophin protein at the myofiber membrane. A method to rescue dystrophin production by antisense oligonucleotides, termed exon-skipping, has been reported for the mdx mouse and in four DMD patients by local intramuscular injection. We sought to test efficacy and toxicity of intravenous oligonucleotide (morpholino)-induced exon skipping in the DMD dog model. METHODS: We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery methods (intramuscular injection and systemic intravenous injection). The efficiency and efficacy of multiexon skipping (exons 6-9) were tested at the messenger RNA, protein, histological, and clinical levels. RESULTS: Weekly or biweekly systemic intravenous injections with a three morpholino cocktail over the course of 5 to 22 weeks induced therapeutic levels of dystrophin expression throughout the body, with an average of about 26% normal levels. This was accompanied by reduced inflammatory signals examined by magnetic resonance imaging and histology, improved or stabilized timed running tests, and clinical symptoms. Blood tests indicated no evidence of toxicity. INTERPRETATION: This is the first report of widespread rescue of dystrophin expression to therapeutic levels in the dog model of DMD. This study also provides a proof of concept for systemic multiexon-skipping therapy. Use of cocktails of morpholino, as shown here, allows broader application of this approach to a greater proportion of DMD patients (90%) and also offers the prospect of selecting deletions that optimize the functionality of the dystrophin protein. PMID- 19288468 TI - Serum biomarker for progranulin-associated frontotemporal lobar degeneration. AB - OBJECTIVE: Mutations that lead to a loss of progranulin (PGRN) explain a considerable portion of the occurrence of frontotemporal lobar degeneration. We tested a biomarker allowing rapid detection of a loss of PGRN. METHODS: We used an enzyme-linked immunosorbent assay to measure in serum the PGRN protein levels of six affected and eight unaffected carriers from within an extended Belgian founder family segregating the null mutation IVS1+5G>C. Further, we measured serum PGRN levels in 2 patients with another null mutation (a Met1 and a frameshift mutation), in 4 patients carrying a predicted pathogenic missense mutation and in 5 patients carrying a benign missense polymorphism, in 9 unaffected noncarrier relatives, and in 22 community controls. RESULTS: Serum PGRN levels were reduced in both affected and unaffected null mutation carriers compared with noncarrier relatives (p(exact) < 0.0001), and allowed perfect discrimination between carriers and noncarriers (sensitivity: 1.0; 1 - specificity: 0.0). Serum PGRN levels in Cys139Arg and Arg564Cys mutation carriers were significantly lower than in controls, but greater than in null mutation carriers, fitting the hypothesis of partial loss of function caused by these missense mutations. As expected, levels for carriers of benign missense polymorphisms were not significantly different from controls. INTERPRETATION: Our results indicate that the serum PGRN level is a reliable biomarker for diagnosing and early detection of frontotemporal lobar degeneration caused by PGRN null mutations, and provided the first in vivo evidence that at least some missense mutations in PGRN may lead to a (partial) loss of PGRN. PMID- 19288469 TI - Cognition and mood in Parkinson's disease in subthalamic nucleus versus globus pallidus interna deep brain stimulation: the COMPARE trial. AB - OBJECTIVE: Our aim was to compare in a prospective blinded study the cognitive and mood effects of subthalamic nucleus (STN) vs. globus pallidus interna (GPi) deep brain stimulation (DBS) in Parkinson disease. METHODS: Fifty-two subjects were randomized to unilateral STN or GPi DBS. The co-primary outcome measures were the Visual Analog Mood Scale, and verbal fluency (semantic and letter) at 7 months post-DBS in the optimal setting compared to pre-DBS. At 7 months post-DBS, subjects were tested in four randomized/counterbalanced conditions (optimal, ventral, dorsal, and off DBS). RESULTS: Forty-five subjects (23 GPi, 22 STN) completed the protocol. The study revealed no difference between STN and GPi DBS in the change of co-primary mood and cognitive outcomes pre- to post-DBS in the optimal setting (Hotelling's T(2) test: p = 0.16 and 0.08 respectively). Subjects in both targets were less "happy", less "energetic" and more "confused" when stimulated ventrally. Comparison of the other 3 DBS conditions to pre-DBS showed a larger deterioration of letter verbal fluency in STN, especially when off DBS. There was no difference in UPDRS motor improvement between targets. INTERPRETATION: There were no significant differences in the co-primary outcome measures (mood and cognition) between STN and GPi in the optimal DBS state. Adverse mood effects occurred ventrally in both targets. A worsening of letter verbal fluency was seen in STN. The persistence of deterioration in verbal fluency in the off STN DBS state was suggestive of a surgical rather than a stimulation-induced effect. Similar motor improvement were observed with both STN and GPi DBS. PMID- 19288470 TI - Bromoxynil degradation in a Mississippi silt loam soil. AB - BACKGROUND: The objectives of these laboratory experiments were: (1) to assess bromoxynil sorption, mineralization, bound residue formation and extractable residue persistence in a Dundee silt loam collected from 0-2 cm and 2-10 cm depths under continuous conventional tillage and no-tillage; (2) to assess the effects of autoclaving on bromoxynil mineralization and bound residue formation; (3) to determine the partitioning of non-extractable residues; and (4) to ascertain the effects of bromoxynil concentration on extractable and bound residues and metabolite formation. RESULTS: Bromoxynil K(d) values ranged from 0.7 to 1.4 L kg(-1) and were positively correlated with soil organic carbon. Cumulative mineralization (38.5% +/- 1.5), bound residue formation (46.5% +/- 0.5) and persistence of extractable residues (T(1/2) < 1 day) in non-autoclaved soils were independent of tillage and depth. Autoclaving decreased mineralization and bound residue formation 257-fold and 6.0-fold respectively. Bromoxynil persistence in soil was rate independent (T(1/2) < 1 day), and the majority of non-extractable residues (87%) were associated with the humic acid fraction of soil organic matter. CONCLUSIONS: Irrespective of tillage or depth, bromoxynil half-life in native soil is less than 1 day owing to rapid incorporation of the herbicide into non-extractable residues. Bound residue formation is governed principally by biochemical metabolite formation and primarily associated with soil humic acids that are moderately bioavailable for mineralization. These data indicate that the risk of off-site transport of bromoxynil residues is low owing to rapid incorporation into non-extractable residues. PMID- 19288471 TI - A sterile-female technique proposed for control of Striga hermonthica and other intractable weeds: advantages, shortcomings and risk management. AB - Weeds have posed intractable challenges to farmers since the dawn of agriculture. This article describes in detail a proposed control strategy based on the introduction of genes conferring female sterility into the genome of an intractable target weed. Spread of these genes through target populations via pollen would be facilitated by their incorporation within active transposable elements. Advantages (e.g. self-dissemination, self-proliferation, target specificity) and shortcomings (e.g. high cost, long project incubation period, limited range of possible targets) of this strategy are discussed in depth, as are assessment and management of its attendant biological and ecological risks, such as the risk of introduced genes spreading to non-target species. The parasitic weed Striga hermonthica (Del.) Benth. is examined as a potential target. PMID- 19288472 TI - Methods for selecting hypervirulent biocontrol agents of weeds: why and how. AB - A considerable number of plant pathogens have been studied for their possible use in weed control. Some have proven virulent enough to control weed species and to compete commercially with chemical herbicides. However, most pathogens of weeds are not useful in their wild form because they are not sufficiently host-specific and/or virulent. The authors believe that these barriers can be overcome. The present research has focused on the inhibitory effects of certain amino acids on the growth and development of specific plants. Pathogens that overproduce these selected amino acids can be easily selected from a pool of spontaneous mutants. Such mutants can have increased pathogenicity to their target weed and enhanced field performance as biocontrol agents. Enhancement of biocontrol efficacy in three separate pathogen-host systems, two with Fusarium and one with Pseudomonas, has already been reported. It is proposed to use the same technology to enhance the biocontrol efficacy of the two species of Fusarium that are host-specific pathogens of the broomrape group of parasitic weeds. The stepwise approach outlined can lead to obtaining enhanced biocontrol agents capable of producing inhibitory levels of selected amino acids in situ. It is proposed that these approaches, in combination with other methods of virulence enhancement, will lead to sustainable systems of biological control of parasitic weeds. PMID- 19288473 TI - Adherence to diagnostic guidelines and quality indicators in asthma and COPD in Swedish primary care. AB - PURPOSE: To study the clinical evaluation and treatment of patients with asthma and COPD in primary care in Sweden, with a focus on adherence to recommended guidelines and quality indicators. METHODS: All visits at health care centres in Skaraborg, Sweden, are documented in computerized medical records constituting the Skaraborg Primary Care Database (SPCD). In a register-based retrospective observational study, all patients diagnosed with asthma or COPD during 2000-2005 (n = 12,328) were identified. In a 5% random sample (n = 623), information on performed investigations at initial visits and at follow-up during 2004-2005 was collected. Compliance with procedures as recommended by national guidelines was used for quality assessment. RESULTS: Among 499 patients with asthma, 167 (33%) were investigated with spirometry or Peak Expiratory Flow (PEF) during initial visits in agreement with guidelines. Correspondingly, 40 out of 124 patients with COPD (32%) were investigated with spirometry. During follow-up, evaluation in agreement with guidelines was performed in 130 (60%) of patients with asthma and in 35 patients out of 77 (45%) with COPD. Prescribing of ICS reached quality target, still every second patient made an acute visit during follow-up. CONCLUSION: Adherence to recommended guidelines in asthma/COPD was low. Acute visits were common and despite the prescribing of ICS according to recommendations, patients still seem uncontrolled in their disease. There is a need for quality improvement in the clinical evaluation and treatment of patients with asthma and COPD. PMID- 19288474 TI - Docking to heme proteins. AB - In silico screening has become a valuable tool in drug design, but some drug targets represent real challenges for docking algorithms. This is especially true for metalloproteins, whose interactions with ligands are difficult to parametrize. Our docking algorithm, EADock, is based on the CHARMM force field, which assures a physically sound scoring function and a good transferability to a wide range of systems, but also exhibits difficulties in case of some metalloproteins. Here, we consider the therapeutically important case of heme proteins featuring an iron core at the active site. Using a standard docking protocol, where the iron-ligand interaction is underestimated, we obtained a success rate of 28% for a test set of 50 heme-containing complexes with iron ligand contact. By introducing Morse-like metal binding potentials (MMBP), which are fitted to reproduce density functional theory calculations, we are able to increase the success rate to 62%. The remaining failures are mainly due to specific ligand-water interactions in the X-ray structures. Testing of the MMBP on a second data set of non iron binders (14 cases) demonstrates that they do not introduce a spurious bias towards metal binding, which suggests that they may reliably be used also for cross-docking studies. PMID- 19288475 TI - Size-dependent cytotoxicity of monodisperse silica nanoparticles in human endothelial cells. AB - The effect that monodisperse amorphous spherical silica particles of different sizes have on the viability of endothelial cells (EAHY926 cell line) is investigated. The results indicate that exposure to silica nanoparticles causes cytotoxic damage (as indicated by lactate dehydrogenase (LDH) release) and a decrease in cell survival (as determined by the tetrazolium reduction, MTT, assay) in the EAHY926 cell line in a dose-related manner. Concentrations leading to a 50% reduction in cell viability (TC(50)) for the smallest particles tested (14-, 15-, and 16-nm diameter) ranging from 33 to 47 microg cm(-2) of cell culture differ significantly from values assessed for the bigger nanoparticles: 89 and 254 microg cm(-2) (diameter of 19 and 60 nm, respectively). Two fine silica particles with diameters of 104 and 335 nm show very low cytotoxic response compared to nanometer-sized particles with TC(50) values of 1095 and 1087 microg cm(-2), respectively. The smaller particles also appear to affect the exposed cells faster with cell death (by necrosis) being observed within just a few hours. The surface area of the tested particles is an important parameter in determining the toxicity of monodisperse amorphous silica nanoparticles. PMID- 19288476 TI - Anti-inflammatory properties of cryptolepine. AB - Cryptolepine is the major alkaloid of the West African shrub, Cryptolepis sanguinolenta. Cryptolepine has been shown to inhibit nitric oxide production, and DNA binding of Nuclear Factor-kappa B following inflammatory stimuli in vitro. In order to validate the anti-inflammatory property of this compound in vivo, we investigated its effects on a number of animal models of inflammation. Cryptolepine (10-40 mg/kg i.p.) produced significant dose-dependent inhibition of the carrageenan-induced rat paw oedema, and carrageenan-induced pleurisy in rats. These effects were compared with those of the non-steroidal anti-inflammatory drug indomethacin (10 mg/kg). At doses of 10-40 mg/kg i.p., cryptolepine inhibited lipopolysaccharide (LPS)-induced microvascular permeability in mice in a dose-related fashion. Oral administration of up to 40 mg/kg of the compound for four consecutive days did not induce gastric lesion formation in rats. Analgesic activity was also exhibited by cryptolepine through a dose-related (10-40 mg/kg i.p.) inhibition of writhing induced by i.p. administration of acetic acid in mice. The results of this study reveal that cryptolepine possesses in vivo anti inflammatory activity. PMID- 19288477 TI - Ultrasound stimulates NF-kappaB activation and iNOS expression via the Ras/Raf/MEK/ERK signaling pathway in cultured preosteoblasts. AB - It has been shown that ultrasound (US) stimulation accelerates fracture healing in the animal models and non-operatively clinical uses. Nitric oxide (NO) is a crucial early mediator in mechanically induced bone formation. Here we found that US-mediated inducible nitric oxide synthase (iNOS) expression was attenuated by Ras inhibitor (manumycin A), Raf-1 inhibitor (GW5074), MEK inhibitor (PD98059), NF-kappaB inhibitor (PDTC), and IkappaB protease inhibitor (TPCK). US-induced Ras activation was inhibited by manumycin A. Raf-1 phosphorylation at Ser(338) by US was inhibited by manumycin A and GW5074. US-induced MEK and ERK activation was inhibited by manumycin A, GW5074, and PD98059. Stimulation of preosteoblasts with US activated IkappaB kinase alpha/beta (IKK alpha/beta), IkappaBalpha phosphorylation, p65 phosphorylation at Ser(276), p65, and p50 translocation from the cytosol to the nucleus, and kappaB-luciferase activity. US-mediated an increase of IKK alpha/beta, IkappaBalpha, and p65 phosphorylation, kappaB luciferase activity and p65 and p50 binding to the NF-kappaB element was inhibited by manumycin A, GW5074, and PD98059. Our results suggest that US increased iNOS expression in preosteoblasts via the Ras/Raf 1/MEK/ERK/IKKalphabeta and NF-kappaB signaling pathways. PMID- 19288478 TI - 5-(Pyrrolidin-2-yl)tetrazole-catalyzed aldol and mannich reactions: acceleration and lower catalyst loading in a continuous-flow reactor. AB - Continuous organocatalysis: Fast aldol and Mannich reactions require less catalyst when conducted in a microreactor. A proline tetrazole derivative (5-10 mol %) catalyzes asymmetric aldol reactions between various aromatic aldehydes and ketones in microreactor at 60 degrees C with reaction times ranging from 10 to 30 min. PMID- 19288479 TI - Identification of a highly efficient alkylated pincer thioimido-palladium(II) complex as the active catalyst in Negishi coupling. AB - The induction period of Negishi coupling catalyzed by pincer thioamide-palladium complex 1 was investigated. A heterogeneous mechanism was excluded by kinetic studies and comparison with Negishi coupling reactions promoted by Pd(OAc)(2)/Bu(4)NBr (a palladium-nanoparticle system). Tetramer 2 was isolated from the reaction of 1 and organozinc reagents. Dissociation of complex 2 by PPh(3) was achieved, and the structure of resultant complex 8 was confirmed by X ray diffraction analysis. A novel alkylated pincer thioimido-Pd(II) complex, 7, generated from catalyst precursor 1 and basic organometallic reagents (RM), was observed by in situ IR, (1)H NMR, and (13)C NMR spectroscopy for the first time. The reaction of 7 with methyl 2-iodobenzoate afforded 74% of the cross-coupled product, methyl 2-methylbenzoate, together with 60% of Pd(II) complex 2. Furthermore, the catalyst, as an electron-rich Pd(II) species, efficiently promoted the Negishi coupling of aryl iodides and alkylzinc reagents without an induction period, even at low temperatures (0 degrees C or -20 degrees C). To evaluate the influence of the catalyst structure upon the induction period, complex 9 was prepared, in which the nBu groups of 1 were displaced by more bulky 1,3,5-trimethylphenyl groups. Trimer 10 was isolated from the reaction of complex 9 and basic organometallic reagents such as CyZnCl or CyMgCl (Cy: cyclohexyl); this is consistent with the result obtained with complex 1. The rate in the induction period of the model reaction catalyzed by 9 was faster than that with 1. Plausible catalytic cycles for the reaction, based upon the experimental results, are discussed. PMID- 19288480 TI - Determination of the catalytic pathway of a manganese arginase enzyme through density functional investigation. AB - The catalytic mechanism of dimanganese-containing arginase enzyme has been investigated by DFT calculations. Two exchange-correlation functionals, B3 LYP and MPWB1 K, have been used to construct the potential energy profiles for the hydrolysis of an arginine substrate performed by an arginase active site model system. Two reaction mechanisms have been investigated, one involving a water molecule (mechanism 1) and the other involving a hydroxide ion (mechanism 2) as nucleophilic agent. Results obtained in the gas phase and in the protein environment have indicated that mechanism 1 involving a water molecule entails structural features as well as an activation energy for the rate-determining step that are inconsistent with experimental data available for the arginase enzyme. On the other hand, when a hydroxide ion is present at the Mn2 site, a lower activation energy and a structural arrangement closer to the experimental indication are obtained. PMID- 19288481 TI - Supramolecular ABC triblock copolymers. AB - Just add it! Ruthenium initiators functionalized with hydrogen-bonding sites were utilized in ring-opening metathesis polymerization to prepare heterotelechelic polymers with hydrogen-bonding and metal-coordination units in a single step. Supramolecular ABC triblock copolymers were then self-assembled in one pot by simply adding complementary telechelic polymers to a solution of the heterotelechelic polymer (see picture). PMID- 19288482 TI - A very efficient cerium(IV) ammonium nitrate catalyzed, four-component synthesis of tetrahydropyridines and its application in the concise generation of functionalized homoquinolizine frameworks. AB - Molecular diversity: A cerium(IV) ammonium nitrate (CAN) catalyzed, four component reaction from very simple acyclic starting materials afforded densely substituted tetrahydropyridines, which were transformed into homoquinolizines by using a gamma-alkylation-ring-closing metathesis (RCM) sequence (see scheme).The cerium(IV) ammonium nitrate (CAN) catalyzed, four-component reaction between primary aliphatic amines, beta-ketoesters or beta-ketothioesters, alpha,beta unsaturated aldehydes, and alcohols provided a very efficient and atom-economical access to substituted 6-alkoxy-2-methyl-1,4,5,6-tetrahydropyridines. These materials were then transformed into homoquinolizine derivatives in excellent yields by using a two-step sequence comprised of regioselective gamma deprotonation-allylation and ring-closing metathesis reactions. The possibility of displacing the alkoxy group by allylsilane nucleophiles, presumably through a vinylogous acyliminium intermediate species, was also demonstrated. PMID- 19288483 TI - Computational and variable-temperature infrared spectroscopic studies on carbon monoxide adsorption on zeolite Ca-A. AB - Carbon monoxide adsorption on LTA (Linde type 5A) zeolite Ca-A is studied by using a combination of variable-temperature infrared spectroscopy and computational methods involving periodic density functional calculations and the correlation between stretching frequency and bond length of adsorbed CO species (nu(CO)/r(CO) correlation). Based on the agreement between calculated and experimental results, the main adsorption species can be identified as bridged Ca(2+)...CO...Ca(2+) complexes formed on dual-cation sites constituted by a pair of nearby Ca(2+) cations. Two types of such species can be formed: One of them has the two Ca(2+) ions located on six-membered rings of the zeolite framework and is characterized by a C-O stretching frequency in the range of 2174-2179 cm( 1) and an adsorption enthalpy of -31 to -33 kJ mol(-1), whereas the other bridged CO species is formed between a Ca(2+) ion located on an eight-membered ring and another one on a nearby six-membered ring and is characterized by nu(CO) in the range 2183-2188 cm(-1) and an adsorption enthalpy of -46 to -50 kJ mol(-1). Ca(2+)...CO monocarbonyl complexes are also identified, and at a relatively high CO equilibrium pressure, dicarbonyl species can also be formed. PMID- 19288484 TI - Hydrogen-atom transfer reactions from ortho-alkoxy-substituted phenols: an experimental approach. AB - The role of intramolecular hydrogen bonding (HB) on the bond-dissociation enthalpy (BDE) of the phenolic O-H and on the kinetics of H-atom transfer to peroxyl radicals (k(inh)) of several 2-alkoxyphenols was experimentally quantified by the EPR equilibration technique and by inhibited autoxidation studies. These compounds can be regarded as useful models for studying the H-atom abstraction from 2-OR phenols, such as many lignans, reduced coenzyme Q and curcumin. The effects of the various substituents on the BDE(O-H) of 2-methoxy, 2 methoxy-4-methyl, 2,4-dimethoxyphenols versus phenol were measured in benzene solution as -1.8; -3.7; -5.4 kcal mol(-1), respectively. In the case of polymethoxyphenols, significant deviations from the BDE(O-H) values predicted by the additive effects of the substituents were found. The logarithms of the k(inh) constants in cumene were inversely related to the BDE(O-H) values, obeying a linear Evans-Polanyi plot with the same slope of other substituted phenols and a y-axis intercept slightly smaller than that of 2,6-dimethyl phenols. In the cases of phenols having the 2-OR substituent included in a five-membered condensed ring (i.e, compounds 9-11), both conformational isomers in which the OH group points toward or away from the oxygen in position 2 were detected by FTIR spectroscopy and the intramolecular HB strength was thus estimated. The contribution to the BDE(O-H) of the ortho-OR substituent in 9, corrected for intramolecular HB formation, was calculated as -5.6 kcal mol(-1). The similar behaviour of cyclic and non-cyclic ortho-alkoxy derivatives clearly showed that the preferred conformation of the OMe group in ortho-methoxyphenoxyl radicals is that in which the methyl group points away from the phenoxyl oxygen, in contrast to the geometries predicted by DFT calculations. PMID- 19288485 TI - Well-defined silica-supported calcium reagents: control of Schlenk equilibrium by grafting. AB - Calcium reagents Ca(alpha-Me(3)Si-2-Me(2)N-benzyl)(2) x 2 thf (1) and Ca[N(SiMe(3))(2)](2) x 2 thf (2) reacted with silica partially dehydroxylated at 700 degrees C to afford materials that bear ([triple bond]SiO)Ca(alpha-Me(3)Si-2 Me(2)N-benzyl) x 1.6 thf (SiO(2)-1) and ([triple bond]SiO)Ca [N(SiMe(3))(2)] x 1.3 thf (SiO(2)-2) fragments, respectively. Due to the bulk of the supported complexes, the silanol groups are only partially metalated: 50% in SiO(2)-1 and 70% in SiO(2)-2. In the case of SiO(2)-2, a parallel SiMe(3)-capping side reaction affords in fine a silanol-free surface. The materials were characterized by IR spectroscopy, 1D and 2D solid-state high-field NMR spectroscopy, and elemental analysis. Reaction of 2 with one equivalent of the bulky silanol (tBuO)(3)SiOH, a silica-surface mimic, afforded the homoleptic bis-silyloxide calcium derivative through ligand exchange (Schlenk equilibrium), and a derivative was isolated and structurally characterized. Preliminary studies have shown that both grafted benzyl and amide derivatives are active in olefin hydrosilylation, intramolecular hydroamination, and styrene polymerization, with evidence showing that catalysis occurs through supported species. In styrene polymerization, a marked influence of the surface acting as a ligand on the stereoselectivity of the reaction was observed, as syndiotactic-rich polystyrene (88% of r diads) was obtained. These results illustrate that grafting of calcium benzyl or amide compounds on a silica surface is a new concept to prevent ligand exchange through the Schlenk equilibrium. Heteroleptic calcium complexes that cannot be synthesized as stable molecular species in solution can be obtained as silica-supported species which have been shown to be catalytically active. PMID- 19288486 TI - A tin-tungsten mixed oxide as an efficient heterogeneous catalyst for C-C bond forming reactions. AB - The tin-tungsten mixed oxide prepared by the calcination of the tin-tungsten hydroxide precursor with a Sn/W molar ratio of 2 at 800 degrees C (SnW2-800) acts as an effective and reusable solid catalyst for C-C bond-forming reactions, such as the cyclization of citronellal, the Diels-Alder reaction, and the cyanosilylation of carbonyl compounds with trimethylsilyl cyanide (TMSCN). Various kinds of structurally diverse aliphatic, aromatic, and unsaturated, heteroatom-containing substrates could be converted into the desired products in high to excellent yields. The observed catalyses for these reactions were truly heterogeneous and the recovered catalyst could be reused several times without an appreciable loss of its high catalytic performance. The Bronsted acid sites generated on the aggregated polytungstate species on SnW2-800 likely play an important role in the C-C bond-forming reactions. PMID- 19288487 TI - Preparation and characterisation of a new inclusion compound of cucurbit[8]uril with a nitroxide radical. PMID- 19288488 TI - Synthesis of pyrimidines and triazines in ice: implications for the prebiotic chemistry of nucleobases. AB - Herein, we report the efficient synthesis of RNA bases and functionalized s triazines from 0.1 M urea solutions in water after subjection to freeze-thaw cycles for three weeks. The icy solution was under a reductive, methane-based atmosphere, which was subjected to spark discharges as an energy source for the first 72 h of the experiment. Analysis of the products indicates the synthesis of the s-triazines cyanuric acid, ammeline, ammelide, and melamine, the pyrimidines cytosine, uracil, and 2,4-diaminopyrimidine, and the purine adenine. An experiment performed as a control at room temperature, with the urea solution in the liquid phase and with the same atmosphere and energy source, led to the synthesis of hydantoins and insoluble tholin, but there was no evidence of the synthesis of pyrimidines or triazines. The synthesis of pyrimidines from urea is possible under a methane/nitrogen atmosphere only at low temperature, in the solid phase. The generation of both pyrimidines and triazines in comparable yields from urea, together with a possible role for triazines as alternative nucleobases, opens new perspectives on the prebiotic chemistry of informational polymers. PMID- 19288489 TI - Density functional theory investigation of the alkyl-alkyl Negishi cross-coupling reaction catalyzed by N-heterocyclic carbene (NHC)-Pd complexes. AB - A novel mechanism is proposed for the Pd-1,3-(2,6-diisopropylphenyl)imidazolyl-2 ylidene (1) catalyzed Negishi reaction. DFT computations supported by atoms-in molecules (AIM) analyses of non-truncated models show that a "steric wall" created by the N-substituent on the ligand guides reactants to and from the Pd center. Notably, transmetalation and not oxidative addition is found to be rate limiting. Additionally, a key Pd-Zn interaction (approximately = 2.4 A, rho(b) approximately = 0.0600 au) is identified in the mechanism. This interaction persists beyond reductive elimination and, in combination with the ligand, facilitates reductive elimination by creating a highly sterically crowded environment in the coordination sphere of the Pd center. PMID- 19288490 TI - Adiabatic low-pass J filters for artifact suppression in heteronuclear NMR. AB - NMR artifact purging: Modern NMR experiments depend on efficient coherence transfer pathways for their sensitivity and on suppression of undesired pathways leading to artifacts for their spectral clarity. A novel robust adiabatic element suppresses hard-to-get-at artifacts (see picture). PMID- 19288491 TI - Flavonoid dimers as bivalent modulators for p-glycoprotein-based multidrug resistance: structure-activity relationships. AB - We recently described the modulatory activities of apigenin homodimers linked by ethylene glycol units in multidrug- resistant breast cancer and leukemic cells overexpressing ABCB1 (P-glycoprotein, P-gp). To further improve the potency of these dimers, a small library of flavonoid homodimers and heterodimers were synthesized, and their in vitro activity in reversing cellular resistance to paclitaxel, along with structure-activity relationships (SAR), were evaluated using a P-gp-expressing human breast cancer cell line. Among these synthesized homodimers, many showed more potent reversing activity than that of the parent compound and verapamil. Two compounds in particular showed promising reversing activity at sub-micromolar concentrations with no cytotoxic effects. Regarding SAR trends, flavonoid dimers with nonpolar and hydrophobic substituents (e.g., methyl and ethyl groups) generally showed more potent resistance-reversing activity than that of dimers with polar and hydrophilic substituents (e.g. hydroxy groups) at the C3, C6, and C7 positions, but not at C5. In terms of substituent steric bulk at C6, it was found that the flavonoid dimer with methyl groups was optimal, with bulkier substituents leading to lower reversing activity. Comparisons of flavonoid heterodimers with the corresponding homodimers revealed that the two binding sites on P-gp for flavonoid moieties are quite similar to each other. Besides paclitaxel, these new compounds also increased drug accumulation and enhanced the cytotoxicity of other cancer drugs such as doxorubicin, vincristine, and vinblastine by decreasing the IC(50) values 4-45 fold. PMID- 19288492 TI - Structure-based optimization of benzoic acids as inhibitors of protein tyrosine phosphatase 1B and low molecular weight protein tyrosine phosphatase. AB - We have optimized previously discovered benzoic acids 1, which are active as inhibitors of PTP1B and LMW-PTP, two protein tyrosine phosphatases that have emerged as attractive targets for the development of novel therapeutic agents for the treatment of diabetes, obesity, and cancer. Our efforts led to the identification of new and more potent analogues with appreciable selectivity toward human PTP1B and the IF1 isoform of human LMW-PTP. PMID- 19288493 TI - Role of RAF/MEK/ERK pathway, p-STAT-3 and Mcl-1 in sorafenib activity in human pancreatic cancer cell lines. AB - Sorafenib is a multikinase inhibitor that has shown promising therapeutic results in different tumor histotypes, both as a single agent or in combination with other treatments. We analyzed the in vitro activity of sorafenib in pancreatic cancer, one of the most lethal and chemo-radio-resistant tumors, using four human pancreatic cancer cell lines (t3m4, Capan 1, Capan 2, and MiaPaca 2), characterized by different K-ras gene status and RAF/MEK/ERK profile. Sorafenib exerted a strong anti-proliferative effect independently of RAS/RAF/MEK/ERK and induced various degrees of apoptosis in the cell lines. The mechanisms involved were explored in detail in t3m4 and Capan 1, in which sorafenib induced the highest and lowest levels of apoptosis, respectively. In t3m4, the RAF/AKT/STAT-3 rather than the RAF/MEK/ERK pathway was involved, whereas in Capan 1 cells there was a strong decrease in pMEK and pERK which was not accompanied by an important reduction in RAF, AKT, and STAT-3 proteins or in their phosphorylation. Moreover, U0126-induced MEK inhibition did not induce apoptosis in any cell line, reinforcing the hypothesis of a MEK/ERK-independent mechanism of sorafenib activity. Mcl-1 appears to play a crucial role in sorafenib-induced apoptosis. In fact, both protein and mRNA were downregulated in t3m4 and upregulated in Capan 1, in which siRNA-induced silencing resulted in the same level of apoptosis as observed in t3m4. Our results show that sorafenib exerts anti-proliferative and pro-apoptotic activity in pancreatic cancer cells. Used singly or in combination with other drugs, it could therefore represent valid treatment for pancreatic cancer. PMID- 19288494 TI - Cell death induced by zinc and cadmium is mediated by clusterin in cultured mouse seminiferous tubules. AB - Sertoli cells, lining the walls of the seminiferous tubules, are in close contact with and regulate all aspects of the development of the germ cells. Clusterin, is a glycoprotein produced abundantly by Sertoli cells, and associated with either apoptosis or cell survival. Zinc is present in high concentrations in the testis and required for sperm development by an as yet unknown mechanism. Permeation of zinc into cells via voltage-gated calcium channels (VGCCs), however, is suggested to induce cell death. We examined the possibility that Zn(2+) acts via clusterin to regulate germ cell survival. Employing an ex vivo model of mouse testis, we have assessed the role of permeation of heavy metal ions on clusterin production and secretion. Up-regulation of clusterin expression and its secretion was observed after a short exposure to zinc or to cadmium under depolarizing conditions. Expression of zinc transporter-1 (ZnT-1), previously shown to regulate Zn(2+) influx, increased following prolonged application of zinc or cadmium to the explants and prevented clusterin up-regulation by subsequent exposure to these ions. Inhibition of the MAPK and PI3K pathways reduced the up regulation of clusterin following the intracellular rise of Zn(2+) or Cd(2+). Neutralization of secreted clusterin by an antibody or attenuation of clusterin up-regulation by inhibition of Zn(2+) permeation via the LTCC, reduced cell death in cultured seminiferous tubule cells. Taken together, our results indicate that Zn(2+) and Cd(2+) influx induce expression and secretion of clusterin, thereby linking metal homeostasis and germ cell fate. PMID- 19288495 TI - Defective co-activator recruitment in osteoclasts from microphthalmia-oak ridge mutant mice. AB - The three basic DNA-binding domain mutations of the microphthalmia-associated transcription factor (Mitf), Mitf(mi/mi), Mitf(or/or), and Mitf(wh/wh) affect osteoclast differentiation with variable penetrance while completely impairing melanocyte development. Mitf(or/or) mice exhibit osteopetrosis that improves with age and their osteoclasts form functional multinuclear osteoclasts, raising the question as to why the Mitf(or/or) mutation results in osteopetrosis. Here we show that Mitf(or/or) osteoclasts express normal levels of acid phosphatase 5 (Acp5) mRNA and significantly lower levels of Cathepsin K (Ctsk) mRNA during receptor activator of nuclear factor kappa B (NFkappaB) ligand (RANKL)-mediated differentiation. Studies using chromatin immunoprecipitation (ChIP) analysis indicate that low levels of Mitf(or/or) protein are recruited to the Ctsk promoter. However, enrichment of Mitf-transcriptional co-activators PU.1 and Brahma-related gene 1 (Brg1) are severely impaired at the Ctsk promoter of Mitf(or/or) osteoclast precursors, indicating that defective recruitment of co activators by the mutant Mitf(or/or) results in impaired Ctsk expression in osteoclasts. Cathepsin K may thus represent a unique class of Mitf-regulated osteoclast-specific genes that are important for osteoclast function. PMID- 19288496 TI - MUC4 involvement in ErbB2/ErbB3 phosphorylation and signaling in response to airway cell mechanical injury. AB - The receptor tyrosine kinases ErbB2 and ErbB3 are phosphorylated in response to injury of the airway epithelium. Since we have shown that the membrane mucin MUC4 can act as a ligand/modulator for ErbB2, affecting its localization in polarized epithelial cells and its phosphorylation, we questioned whether Muc4 was involved, along with ErbB2 and ErbB3, in the damage response of airway epithelia. To test this hypothesis, we first examined the localization of MUC4 in human airway samples. Both immunocytochemistry and immunofluorescence showed a co localization of MUC4 and ErbB2 at the airway luminal surface. Sequential immunoprecipitation and immunoblotting from airway cells demonstrated that the MUC4 and ErbB2 are present as a complex in airway epithelial cells. To assess the participation of MUC4 in the damage response, cultures of NCI-H292 or airway cells were scratch-wounded, then analyzed for association of phospho-ErbB2 and ErbB3 with MUC4 by sequential immunoprecipitation and immunoblotting. Wounded cultures exhibited increased phosphorylation of both receptors in complex with MUC4. Scratch wounding also increased activation of the downstream pathway through Akt, as predicted from our previous studies on Muc4 effects on ErbB2 and ErbB3. The participation of MUC4 in the phosphorylation response was also indicated by siRNA repression of MUC4 expression, which resulted in diminution of the phosphorylation of ErbB2 and ErbB3. These studies provide a new model for the airway epithelial damage response, in which the MUC4-ErbB2 complex is a key element in the sensor mechanism and phosphorylation of the receptors. PMID- 19288497 TI - Respiratory symptoms and lung function in foundry workers exposed to low molecular weight isocyanates. AB - BACKGROUND: This study investigated the prevalence of ocular and respiratory symptoms and lung function in foundry workers exposed to isocyanic acid (ICA) and methyl isocyanate (MIC). METHODS: Foundry workers (n = 64) using the Hot Box binder technique and referents (n = 134) completed two questionnaires and a spirometry before and after a day shift. Findings were analyzed relating to exposure measurements of ICA, MIC, and formaldehyde. RESULTS: Exposed workers reported more frequently ocular irritation and coughing. Pre-shift lung function was lower than predicted in both groups of workers, and was not associated with exposure duration. Decrease in lung function over shift was more pronounced in exposed workers, but this was not associated with quantitative measures of the exposures. CONCLUSIONS: An increased prevalence of ocular and respiratory symptoms, as well as a small decrease in lung function over shift, was observed among exposed workers. No associations with exposure to monoisocyanates and formaldehyde were seen, thus suggesting that other irritants in the foundry environment might be the cause. The effects seemed to be small and not clinically relevant on an individual level. Further studies using a follow-up design may be warranted. PMID- 19288498 TI - Inorganic phosphate modulates responsiveness to 24,25(OH)2D3 in chondrogenic ATDC5 cells. AB - Chondrogenic ATDC5 cells were used as a model of in vitro endochondral maturation to study the role of inorganic phosphate (Pi) in the regulation of growth plate chondrocytes by vitamin D3 metabolites. ATDC5 cells that were cultured for 10 days post-confluence in differentiation media and then treated for 24 h with Pi produced a type II collagen matrix based on immunohistochemistry and expressed mRNAs for several chondrocytic markers, including aggrecan, collagen types II and X, cartilage oligomeric matrix protein, and SOX9. Pi also caused a decrease in [(35)S]-sulfate incorporation and stimulated apoptosis, as evidenced by increased DNA fragmentation and caspase-3 activity. In addition, treatment with Pi induced sensitivity to 24,25-dihydroxyvitamin D3 and this effect was both dose-dependent and was blocked by phosphonoformic acid (PFA), a specific inhibitor of sodium dependent type III Pi transporters. Treatment with 24R,25(OH)(2)D(3) reduced cell number and increased alkaline phosphatase specific activity in a dose-dependent manner. Moreover, 24R,25(OH)(2)D(3) reversed the Pi-induced decrease in incorporation of [(3)H]-thymidine and [(35)S]-sulfate incorporation, as well as the Pi-induced increase in apoptosis. These results suggest that Pi acts as an early chondrogenic differentiation factor, inducing response to 24R,25(OH)(2)D(3); treatment of committed chondrocytes with Pi induces apoptosis, but 24R,25(OH)(2)D(3) mitigates these effects, indicating a possible inhibitory feedback loop. PMID- 19288499 TI - The effect of Echinacea purpurea (L.) Moench extract on experimental prostate hyperplasia. AB - The aim of this study was to examine the effect of purple coneflower (Echinacea purpurea L. Moench) on the prostate gland of rats using an experimental model of benign prostate hyperplasia (BPH). The animals were administered 50 mg/kg of extract preparation for 4 and 8 weeks and the prostate mass and structural degenerative changes were evaluated in the course of the experiment. The administration of E. purpurea extract to rats with hyperplasia for 4 and 8 weeks gradually and significantly reduced the prostate mass and reversed the degenerative changes in the structure of the prostate gland. The present investigation suggests extract of purple coneflower prevents the development of BPH. PMID- 19288501 TI - Cholesterol depletion induces anoikis-like apoptosis via FAK down-regulation and caveolae internalization. AB - Caveolae (lipid rafts), microdomains of the plasma membrane, are known to contain various signalling molecules and consequently are involved in the regulation of many biological functions. To investigate the role of the caveolae in cell survival and adhesion, we disrupted the caveolae by depletion of cholesterol, a major lipid component of the caveolae, with methyl-beta cyclodextrin (MbetaCD) treatment of A431 cells. We found that cholesterol depletion induced an anoikis like cell death involving actin reorganization, resulting in a decrease in cell spreading and an increase in cell detachment, which was reversed by cholesterol addition. Disruption of caveolae led to the down-regulation of FAK, Src activation, tyrosine phosphorylation of caveolin-1 and mobilization of caveolae markers, GM1 and caveolin-1, from the cell surface to the cytoplasm, which were also recovered by cholesterol addition. The expression of dominant-active FAK was able to delay caveolae internalization and apoptosis and attenuated Akt inactivation by MbetaCD, whereas dominant-negative FAK expression resulted in enhanced apoptosis. Moreover, FAK down-regulation by si-RNA resulted in Akt inactivation and thus increased cell death by MbetaCD treatment. Our results suggest that the cholesterol content and/or surface levels of the caveolae affect the activity of FAK, which in turn regulates caveolae internalization and cell survival. PMID- 19288502 TI - Inhibition of lung metastasis in mice by oligonol. AB - Oligonol is a polyphenol formulation enriched with catechin-type oligomers. As an initial approach to assess the chemopreventive potential of Oligonol, the study investigated the effects of Oligonol on the inhibition of lung metastasis induced by B16F-10 melanoma cells in C57BL/6 mice. Oligonol, which is abundantly found in plants, vegetables and fruits, was found to possess antimetastatic activity against B16F-10 melanoma cells. Continued consumption of Oligonol, even at high doses, for long periods does not seem to cause any toxic symptoms because excess Oligonol is stored in adipose tissue rather than in the liver. However, the mechanism by which Oligonol exerts its antimetastatic activity remains unclear. Further investigations are required to clarify the exact role of Oligonol in such B16F-10 melanoma regulation. PMID- 19288503 TI - Carbanion or amide? First charge density study of parent 2-picolyllithium. AB - The negative charge originating from deprotonation of the methyl group is distributed over the 2-picolyl ring. Bonding properties derived from the electron density distribution support the enamide character of picolyllithium (PicLi; the picture shows the deformation density of [2-PicLi x PicH](2)), but electrophilic attack occurs at the deprotonated C atom. This reactivity is rationalized by the electrostatic potential, which guides electrophiles towards the nucleophilic C atom. PMID- 19288504 TI - Iron-catalyzed oxidative heterocoupling between aliphatic and aromatic organozinc reagents: a novel pathway for functionalized aryl-alkyl cross-coupling reactions. AB - Aryl-alkyl cross-coupling products are obtained by the iron-catalyzed oxidative heterocoupling of organozinc reagents under mild conditions. This novel reaction pathway is versatile, allowing for the use of primary and secondary aliphatic diorganozinc reagents as coupling partners as well as tolerating functionalized aryl- and alkylzinc reagents. PMID- 19288505 TI - Exploring the limits of Forster theory for energy transfer at a separation of 20 A. AB - Cutting the corner: An excellent agreement has been obtained between experimental and computed coulombic coupling matrix elements for donor-spacer-acceptor systems, which consist of a boron dipyrromethane donor and acceptor in various stages of protonation. This correlation occurs in spite of reservations about the validity of Forster theory being applied to intramolecular electronic energy transfer (ET) over short (e.g., 20 A) distances (see picture). PMID- 19288506 TI - Incorporation of Fe3O4 nanoparticles into organometallic coordination polymers by nanoparticle surface modification. AB - Surface-modified Fe(3)O(4) nanoparticles (NPs) can be obtained by substituting [(eta(5)-semiquinone)Mn(CO)(3)] for oleylamine surface protecting groups. The resulting NP can function as a nucleus or template to generate crystalline coordination polymers that contain superparamagnetic Fe(3)O(4) NPs. Hybridized magnetic properties can be obtained by introducing paramagnetic metal nodes, such as Mn(2+), into the polymers (see picture). PMID- 19288507 TI - A detection system based on giant magnetoresistive sensors and high-moment magnetic nanoparticles demonstrates zeptomole sensitivity: potential for personalized medicine. AB - Zeptomole detector: A highly sensitive giant-magnetoresistive chip and FeCo nanoparticles can be used to linearly detect 600-4500 copies of streptavidin. Under unoptimized conditions, this system also detects human IL-6 with a sensitivity 13-times higher than that of standard ELISA techniques. PMID- 19288508 TI - Peptides recognize photoresponsive targets. AB - Clones on film: 7-mer peptides that bind to azobenzene-containing polymer films were selected from a phage library under visible light. Isolated clones showed greater affinities to the films under ultraviolet light than those under visible light. Furthermore, the peptide binding responds to the photoinduced isomerization of azobenzene groups. PMID- 19288509 TI - Direct alkenylation of indoles with alpha-oxo ketene dithioacetals: efficient synthesis of indole alkaloids meridianin derivatives. AB - Let's make 'meri': Metal-free direct alkenylation of indoles was realized by acid mediated substitution reactions of alpha-oxo ketene dithioacetals with indoles in trifluoroacetic acid/dichloromethane, selectively affording beta-indolyl mono- and disubstituted alpha,beta-unsaturated carbonyl compounds (see scheme). Condensation of the indolyl/ketene monothioacetals and guanidine nitrate efficiently produced meridianin derivatives. PMID- 19288510 TI - A preferred disrotatory 4n electron Mobius aromatic transition state for a thermal electrocyclic reaction. AB - Not forbidden: Thermal 4n electron electrocyclic reactions of Huckel topology structures proceed via "allowed" conrotatory pathways. However, for a Mobius topology, the Woodward-Hoffmann rules may be reversed and a "forbidden" disrotatory pathway can be preferred as shown theoretically for dodecahexaene 1 that transforms via a Heilbronner-Mobius aromatic transition structure 2 into a cyclic polyene 3. PMID- 19288511 TI - A general approach to the synthesis of gold-metal sulfide core-shell and heterostructures. AB - Cores and effect: Water-dispersible core-shell structures and heterostructures incorporating gold nanocrystals of different shapes (polyhedra, cubes, and rods) and a variety of transition metal sulfide semiconductors (ZnS, CdS, NiS, Ag(2)S, and CuS) are synthesized using cetyltrimethylammonium bromide-encapsulated gold nanocrystals and metal thiobenzoates as starting materials. PMID- 19288512 TI - Compartmentalization of epidermal growth factor receptor in liver plasma membrane. AB - We have investigated epidermal growth factor (EGF)-induced compartmentalization and activation of the EGF receptor (EGFR) in rat liver plasma membrane (PM) raft subfractions prepared by three different biochemical methods previously developed to characterize the composition of membrane rafts. Only detergent-resistant membranes (DRMs) possessed the basic characteristics attributed to membrane rafts. Following the administration of a low dose of EGF (1 microg/100 g BW) the content of EGFR in PM-DRMs did not change significantly; whereas after a higher dose of EGF (5 microg/100 g BW) we observed a rapid and marked disappearance of EGFR (around 80%) from both PM and DRM fractions. Interestingly, following the administration of either a low or high dose of EGF, the pool of EGFR in the PM DRM fraction became highly Tyr-phosphorylated. In accordance with the higher level of EGFR Tyr-Phosphorylation, EGF induced an augmented recruitment of Grb2 and Shc proteins to PM-DRMs compared with whole PM. Furthermore neither high nor low doses of EGF affected the caveolin content in DRMs and PM. These observations suggest that EGFR located in DRMs are competent for signaling, and non-caveolae PM rafts are involved in the compartmentalization and internalization of the EGFR. PMID- 19288513 TI - Metagenomics: Concept, methodology, ecological inference and recent advances. AB - Microorganisms constitute two third of the Earth's biological diversity. As many as 99% of the microorganisms present in certain environments cannot be cultured by standard techniques. Culture-independent methods are required to understand the genetic diversity, population structure and ecological roles of the majority of organisms. Metagenomics is the genomic analysis of microorganisms by direct extraction and cloning of DNA from their natural environment. Protocols have been developed to capture unexplored microbial diversity to overcome the existing barriers in estimation of diversity. New screening methods have been designed to select specific functional genes within metagenomic libraries to detect novel biocatalysts as well as bioactive molecules applicable to mankind. To study the complete gene or operon clusters, various vectors including cosmid, fosmid or bacterial artificial chromosomes are being developed. Bioinformatics tools and databases have added much to the study of microbial diversity. This review describes the various methodologies and tools developed to understand the biology of uncultured microbes including bacteria, archaea and viruses through metagenomic analysis. PMID- 19288514 TI - High-throughput system for determining dissolution kinetics of inclusion bodies. AB - Efficient solubilization is a crucial step during inclusion body processing and dissolving conditions were usually empirically established. Here we describe a new methodology for rapid screening of solubilization conditions and evaluation of dissolution kinetics in microtiter plates. Increase of protein in solution over time was directly related to decrease of turbidity measured by absorbance at 600 nm. Dissolution kinetics of inclusion bodies were described by a first-order reaction kinetics, which was used for drug dissolution modeling. Reaction constants were in the range of 0.01-0.03 s(-1) for buffer conditions providing sufficient solubilization power. This method is not limited to the screening of optimal buffer conditions for solubilization and can be applied for studying other parameters involved in the solubility of IBs, such as pI of the protein, influence of fermentation conditions, influence of initial protein concentration, and more. PMID- 19288515 TI - SCL-LCL-PHA copolymer production by a local isolate, Pseudomonas aeruginosa MTCC 7925. AB - A five-level-four-factor central composite rotary design (CCRD) was employed in combination with response surface methodology (RSM) to optimize the process variables for the production of a novel copolymer consisting of short-chain length (SCL) and long-chain-length (LCL) PHA units, i.e., P(3HB-3HV-3HHD-3HOD) copolymer in Pseudomonas aeruginosa MTCC 7925. The four variables involved in this study were ethanol, glucose, ammonium nitrate (NH(4)NO(3)), and potassium dihydrogen phosphate (KH(2)PO(4)). A second-order polynomial equation was obtained by multiple regression analysis using RSM. The statistical analyses of the results showed that all the four variables had significant impact on the copolymer yield. The model predicted a maximum yield of 81.1% of dry cell weight (dcw) on setting the concentrations of ethanol and glucose at 1.5 and 1.1%, and KH(2)PO(4) and NH(4)NO(3) at 2.79 and 1.86 g/L, respectively. Verification of the predicted value resulted into a yield of 77.6% (dcw). This novel copolymer exhibited comparable material properties with polypropylene (PP) and low density polyethylene (LDPE), thus advocating its potential applications in various fields. PMID- 19288516 TI - Rapid and label-free bacteria detection by surface plasmon resonance (SPR) biosensors. AB - Surface Plasmon Resonance (SPR) biosensor technology has been successfully used for the detection of various analytes such as proteins, drugs, DNA, and microorganisms. SPR-based immunosensors that coupled with a specific antigen antibody reaction, have become a promising tool for the quantification of bacteria as it offers sensitive, specific, rapid, and label-free detection. In this paper, we review the important issues in the development of SPR-based immunoassays for bacteria detection, concentrating on instrumentation, surface functionalization, liquid handling, and surface regeneration. In addition, this review touches on the recent advances in SPR biosensing for sensitivity enhancement. PMID- 19288517 TI - A high mobility group B-1 box A peptide combined with an artery wall binding peptide targets delivery of nucleic acids to smooth muscle cells. AB - The TAT-high mobility group box-1 A box peptide (TAT-HMGB1A) has been reported previously to be able to deliver DNA into cells without cytotoxicity. In this study, an artery wall smooth muscle cell-targeting carrier was developed using TAT-HMGB1A combined with an artery wall binding peptide (ABP). For the production of ABP linked TAT-HMGB1A (TAT-HMGB1A-ABP), pET15b-TAT-HMGB1A-ABP was constructed by inserting the ABP cDNA into pET15b-TAT-HMGB1A. TAT-HMGB1A-ABP was expressed in E. coli and purified by Nickel chelate chromatography. Gel retardation assays showed that TAT-HMGB1A-ABP formed a complex with the plasmid at or above a 5:1 weight ratio (peptide:plasmid). At a 20:1 weight ratio, the zeta-potential was approximately 25 mV and the particle size was approximately 120 nm. TAT-HMGB1A ABP had the highest transfection efficiency in A7R5 smooth muscle cells at a weight ratio of 20:1. TAT-HMGB1A-ABP exhibited higher transfection efficiency in A7R5 cells than PLL or TAT-HMGB1A, while TAT-HMGB1A-ABP had lower transfection efficiencies in Hep3B hepatoma, 293 kidney, NIH3T3 fibroblast, and Raw264.7 macrophage cells compared with PLL. Together, these results suggest that the ABP moiety of the peptide increased transfection efficiency specifically in smooth muscle cells. In a competition assay, the transfection efficiency of TAT-HMGB1A ABP in A7R5 cells was reduced by the addition of free ABP. MTT assays showed that TAT-HMGB1A-ABP did not produce any cytotoxicity in A7R5 cells. Therefore, TAT HMGB1A-ABP may be useful for a targeting gene delivery to smooth muscle cells. PMID- 19288518 TI - Pharmacologic intervention targeting glycolytic-related pathways protects against retinal injury due to ischemia and reperfusion. AB - Retinal ischemia contributes to multiple ocular diseases while aminoguanidine (AMG) treatment significantly inhibits the neuronal and vascular degeneration due to acute retinal ischemia and reperfusion (I/R) injury. In the present study, 2-D DIGE was applied to profile global protein expression changes due to retinal I/R injury, and the protection effects mediated by AMG. Retinal ischemia was induced by elevated intraocular pressure to 80-90 mmHg for 2 h, and reperfusion was established afterward. Retinal tissues were collected 2 days after I/R injury. After 2-D DIGE analysis, a total of 96 proteins were identified. Among them, 28 proteins were identified within gel spots whose intensities were normalized by AMG pretreatment, pathway analysis indicated that most were involved in glycolysis and carbohydrate metabolism. Selected enzymes identified by MS/MS within these pathways, including transketolase, triosephosphate isomerase 1, aldolase C, total enolase, and pyruvate kinase were validated by quantitative Western blots. Glycolytic enzymes and other differentially regulated proteins likely play previously unrecognized roles in retinal degeneration after I/R injury, and inhibition of the resulting metabolic changes, using pharmacologically agents such as AMG, serve to inhibit the changes in metabolism and mitigate retinal degeneration. Select glycolytic enzymes may provide novel therapeutic targets for inhibiting the neuronal and vascular degeneration after retinal I/R injury. PMID- 19288519 TI - Profile of native N-linked glycan structures from human serum using high performance liquid chromatography on a microfluidic chip and time-of-flight mass spectrometry. AB - Protein glycosylation involves the addition of monosaccharides in a stepwise process requiring no glycan template. Therefore, identifying the numerous glycoforms, including isomers, can help elucidate the biological function(s) of particular glycans. A method to assess the diversity of the N-linked oligosaccharides released from human serum without derivatization has been developed using on-line nanoLC and high resolution TOF MS. The N-linked oligosaccharides were analyzed with MALDI FT-ICR MS and microchip LC MS (HPLC Chip/TOF MS). Two microfluidic chips were employed, the glycan chip (40 nL enrichment column, 43 x 0.075 mm(2) i.d. analytical column) and the high capacity chip (160 nL enrichment column, 140 x 0.075 mm(2) i.d. analytical column), both with graphitized carbon as the stationary phase. Both chips offered good sensitivity and reproducibility in separating a heterogeneous mixture of neutral and anionic oligosaccharides between injections. Increasing the length and volume of the enrichment and the analytical columns improved resolution of the peaks. Complex type N-linked oligosaccharides were the most abundant oligosaccharides in human serum accounting for approximately 96% of the total glycans identified, while hybrid and high mannose type oligosaccharides comprise the remaining approximately 4%. PMID- 19288520 TI - The perinucleolar compartment. AB - The perinucleolar compartment (PNC) is a distinct nuclear body that localizes to the nucleolar periphery. The PNC is predominantly found in cancer cells, and recent evidence suggests that PNC prevalence can be a pan-cancer marker for tumors of solid tissue origin. The PNC is a heritable structure enriched with newly transcribed pol III RNAs and RNA-binding proteins, which exchange rapidly with the surrounding nucleoplasm. The structural integrity of the PNC is dependent upon the continuous transcription of pol III RNA and an intact DNA structure. Although the complete structure and function of the PNC remains to be resolved, much progress has been made in the characterization of the PNC in recent years. Here we summarize our current understanding of the dynamics, structure and function of the PNC. PMID- 19288521 TI - Cytogenetic investigation of chromosomal aberrations in cells treated with plantamajoside from Plantago asiatica. AB - Plantago asiatica is a member of the Plantaginaceae family, and is widely distributed in East Asia. In our previous work, a single active compound, plantamajoside was isolated and confirmed to have glycation inhibitory activity, and did not possess toxicity during a 90 day repeated oral toxicity test in rats. In the present study, a chromosomal aberration test was performed to investigate the genotoxicity of plantamajoside. From the results of the cytotoxicity test, plantamajoside proved to be less toxic when it was treated combined with S9 cell fractions. However, there was a significant increase in structural aberrations during the short-term treatment of plantamajoside at its highest dose (5000 microg/mL) even when combined with S9. This seems to have been a natural phenomenon due to the very high dose of plantamajoside that was used. However, to confirm the safety of plantamajoside for its potential use as a phytochemical agent in health products, additional mutagenicity tests are necessary. PMID- 19288522 TI - Amelioration of adjuvant induced arthritis by apocynin. AB - The premise of the study was to investigate the antiarthritic potential of apocynin (APO) in Balb/c mice (in vivo). The experiment showed a dose-dependent decrease in oedema and showed a suppression of proinflammatory cytokines such as TNF-alpha and IL-1beta and mediators such as prostaglandin E(2) (PGE(2)) and LTB(4). At oral doses of 0.5, 1.0, 2.0 and 4.0 mg/kg once daily during the course of the experiment, APO induced an inhibition of T cell mediated immune response causing suppression of CD4+ and CD8+ T cells and of intracellular interferon gamma (IFN-gamma) by flow cytometry in arthritic mice. In parallel there was a dose-dependent inhibition in vascular permeability causing an inhibition in the migration of leucocytes and exudate volume at the site of the inflammatory reaction. These observations validate the immunoregulatory potential of apocynin. PMID- 19288523 TI - A hierarchical MS2/MS3 database search algorithm for automated analysis of phosphopeptide tandem mass spectra. AB - A novel hierarchical MS(2)/MS(3) database search algorithm has been developed to analyze MS(2)/MS(3) phosphopeptides proteomic data. The algorithm is incorporated in an automated database search program, MassMatrix. The algorithm matches experimental MS(2) spectra against a supplied protein database to determine candidate peptide matches. It then matches the corresponding experimental MS(3) spectra against those candidate peptide matches. The MS(2) and MS(3) spectra are used in concert to arrive at peptide matches with overall higher confidence rather than combining MS(2) and MS(3) data searched separately. Receiver operating characteristic analysis showed that hierarchical MS(2)/MS(3) database searches with MassMatrix had better sensitivity and specificity than the two stage MS(2)/MS(3) database searches obtained with MassMatrix, MASCOT, and X!Tandem. A greater number of true peptide matches at a given false rate were identified by use of this new algorithm for data collected on both LCQ and LTQ FTICR mass spectrometers. The additional MS(3) spectral data also improved the overall reliability and the number of true positives (TPs) due to the fact that the TPs of the MS(2)/MS(3) search results had higher scores than those of the MS(2). PMID- 19288524 TI - Highly stable trypsin-aggregate coatings on polymer nanofibers for repeated protein digestion. AB - A stable and robust trypsin-based biocatalytic system was developed and demonstrated for proteomic applications. The system utilizes polymer nanofibers coated with trypsin aggregates for immobilized protease digestions. After covalently attaching an initial layer of trypsin to the polymer nanofibers, highly concentrated trypsin molecules are crosslinked to the layered trypsin by way of a glutaraldehyde treatment. This process produced a 300-fold increase in trypsin activity compared with a conventional method for covalent trypsin immobilization, and proved to be robust in that it still maintained a high level of activity after a year of repeated recycling. This highly stable form of immobilized trypsin was resistant to autolysis, enabling repeated digestions of BSA over 40 days and successful peptide identification by LC-MS/MS. This active and stable form of immobilized trypsin was successfully employed in the digestion of yeast proteome extract with high reproducibility and within shorter time than conventional protein digestion using solution phase trypsin. Finally, the immobilized trypsin was resistant to proteolysis when exposed to other enzymes (i.e., chymotrypsin), which makes it suitable for use in "real-world" proteomic applications. Overall, the biocatalytic nanofibers with trypsin aggregate coatings proved to be an effective approach for repeated and automated protein digestion in proteomic analyses. PMID- 19288525 TI - Identification of proteins of Neisseria meningitidis induced under iron-limiting conditions using the isobaric tandem mass tag (TMT) labeling approach. AB - Isobaric labeling reagents such as Tandem Mass Tags (TMT(R)) enable the genome wide quantification of protein expression levels under different conditions using a gel-free MS/MS-based approach. Here, we applied a TMTduplex approach with two isobaric tags to study the response of the human pathogen Neisseria meningitidis to deprivation of iron, a condition met in the human body. In total, 609 proteins were identified in samples of three independent growth experiments, in which we compared cultures grown in the presence and absence of iron. Expression of 35 proteins was found to be induced or repressed under iron-limiting conditions, including 11 proteins whose ORFs were not previously identified in DNA array studies as being regulated by iron availability at the transcriptional level. These 11 proteins include proteins likely involved in iron metabolism. PMID- 19288526 TI - Subcellular proteomics of Trypanosoma cruzi reservosomes. AB - Reservosomes are the endpoint of the endocytic pathway in Trypanosoma cruzi epimastigotes. These organelles have the particular ability to concentrate proteins and lipids obtained from medium together with the main proteolytic enzymes originated from the secretory pathway, being at the same time a storage organelle and the main site of protein degradation. Subcellular proteomics have been extensively used for profiling organelles in different cell types. Here, we combine cell fractionation and LC-MS/MS analysis to identify reservosome-resident proteins. Starting from a purified reservosome fraction, we established a protocol to isolate reservosome membranes. Transmission electron microscopy was applied to confirm the purity of the fractions. To achieve a better coverage of identified proteins we analyzed the fractions separately and combined the results. LC-MS/MS analysis identified in total 709 T. cruzi-specific proteins; of these, 456 had predicted function and 253 were classified as hypothetical proteins. We could confirm the presence of most of the proteins validated by previous work and identify new proteins from different classes such as enzymes, proton pumps, transport proteins, and others. The definition of the reservosome protein profile is a good tool to assess their molecular signature, identify molecular markers, and understand their relationship with different organelles. PMID- 19288527 TI - Global transcriptome analysis in mouse calvarial osteoblasts highlights sets of genes regulated by modeled microgravity and identifies a "mechanoresponsive osteoblast gene signature". AB - Mechanical unloading is known to be detrimental for the skeleton, but the underlying molecular mechanisms are not fully elucidated. We performed global transcriptome analysis of mouse calvarial osteoblasts grown for 5 days at unit gravity (1g) or under modeled microgravity (0.008g) in the NASA-developed rotating wall vessel (RWV) bioreactor. Elaboration of gene profiling data evidenced that, among the >20,000 gene probes evaluated, 45 genes were significantly up-regulated (cut-off >2) and 88 were down-regulated (cut-off <0.5) in modeled microgravity versus 1g. This set of regulated genes includes genes involved in osteoblast differentiation, function, and osteoblast-osteoclast cross talk, as well as new genes not previously correlated with bone metabolism. Microarray data were validated for subsets of genes by real-time RT-PCR, Western blot, or functional analysis. The significantly modulated genes were then clustered using the GOTM (Gene Ontology Tree Machine) software. This analysis evidenced up-regulation of genes involved in the induction of apoptosis, in response to stress and in the activity of selected growth factors. Other molecular functions, such as extracellular matrix structural constituent, glycosaminoglycan/heparin-binding activity, and other growth factor activity, were instead down-regulated. We finally matched our transcriptome results with other public global gene profiles obtained in loading and unloading conditions, identifying 10 shared regulated genes which could represent an "osteoblast mechanoresponsive gene signature." PMID- 19288528 TI - Neuroactive flavonoid glycosides from Tilia petiolaris DC. extracts. AB - Pharmacological assay guided purification of an ethanol extract of Tilia petiolaris DC. inflorescences resulted in the isolation and identification of isoquercitrin (ISO), quercetin 3-O-glucoside-7-O-rhamnoside (QUE) and kaempferol 3-O-glucoside-7-O-rhamnoside (KAE). The behavioral actions of these glycosylated flavonoids were examined in the hole board, locomotor activity and thiopental induced loss of righting reflex tests in mice. QUE (10 and 30 mg/kg) and KAE (30 mg/kg), intraperitoneally (i.p.) administered to mice, reduced all the parameters measured in the hole board test, but ISO (30 mg/kg) only reduced the number of rearings. Meanwhile QUE at 30 mg/kg i.p. also decreased the ambulatory locomotor activity and increased the sodium thiopental-induced time of loss of the righting reflex suggesting a clear depressant action. The above results demonstrate the occurrence of neuroactive flavonoid glycosides in Tilia. PMID- 19288529 TI - Genistein and curcumin suppress epidermal growth factor-induced MUC5AC mucin production and gene expression from human airway epithelial cells. AB - This study investigated whether genistein and curcumin affect epidermal growth factor (EGF)-induced MUC5AC mucin production and gene expression from human airway epithelial cells. Confluent NCI-H292 cells were pretreated with each agent for 30 min and then stimulated with EGF for 24 h. The MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. The results were as follows: (1) genistein and curcumin inhibited the production of MUC5AC mucin protein induced by EGF, dose-dependently; (2) genistein and curcumin also inhibited the expression of MUC5AC mucin gene induced by EGF. This result suggests that genistein and curcumin can regulate mucin gene expression and production of mucin protein induced by EGF, by directly acting on airway epithelial cells. PMID- 19288531 TI - Propensity scores. PMID- 19288530 TI - Reproductive assessment of hydroalcohol extract of Calendula officinalis L. in Wistar rats. AB - The present study was conducted to evaluate the effects of the administration of a hydroalcohol extract of Calendula officinalis L. flowers (HAE) on the reproductive function of Wistar rats. Four groups of adult male rats were treated orally with HAE at doses of 0, 0.25, 0.5 and 1.0 g/kg for 60 consecutive days. From day 53 to 60 of treatment, rats were mated with untreated and fertile female rats. Reproductive parameters including testicular morphology, reproductive organ weights, fertility index and offspring viability were evaluated. In another protocol, groups of pregnant rats were treated orally with the same doses of HAE from days 1 to 6 (preimplantation period), 7 to 14 (organogenic period) or 15 to 19 (fetal period) of pregnancy. On day 20 of pregnancy, rats were killed for evaluation of maternal and fetal parameters. The results showed that the treatment with HAE did not affect male reproductive parameters. Besides, it was non-toxic in the preimplantation and organogenic periods of pregnancy. However, the HAE induced a decrease of the maternal weight gain when administered during the fetal period. In conclusion, the HAE did not affect male fertility nor had toxic effects in early and middle periods of pregnancy. However, the HAE caused maternal toxicity when administered during the fetal period of pregnancy. PMID- 19288532 TI - Comments on 'Choice of time scale and its effect on significance of predictors in longitudinal studies' by Michael J. Pencina, Martin G. Larson and Ralph B. D'Agostino, Statistics in Medicine 2007; 26:1343-1359. PMID- 19288533 TI - Comparative clinical trial of castor oil and diclofenac sodium in patients with osteoarthritis. AB - A randomized, double-blind, comparative clinical study was conducted to compare the safety and efficacy of castor oil with diclofenac sodium in patients with knee osteoarthritis. Subjects with symptoms of knee osteoarthritis were given a castor oil capsule 0.9 mL (n = 50) thrice daily for 4 weeks or a capsule of diclofenac sodium (n = 50), 50 mg thrice daily for 4 weeks. The subjects completed an overall evaluation of symptom relief at 2 weeks and 4 weeks of completed treatment. The subjects were evaluated by clinical, routine laboratory and radiographic investigations for improvement of disease conditions and also for adverse drug reaction. On completion of 4 weeks treatment it was observed that both drugs were significantly effective in the treatment of knee osteoarthritis (p < 0.001) and adverse drug reactions were high with diclofenac sodium, whereas with castor oil there were no adverse effects reported. The present study indicates that castor oil can be used as an effective therapy in primary knee osteoarthritis. PMID- 19288534 TI - Antibacterial activity of sophoraflavanone G isolated from the roots of Sophora flavescens against methicillin-resistant Staphylococcus aureus. AB - In this study, sophoraflavanone G obtained from Sophora flavescens was evaluated against 10 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), either alone or in combination with ampicillin or oxacillin, via checkerboard assay. At the end point of an optically clear well, the minimum inhibitory concentrations (MICs) ranged from 0.5 to 8 microg/ml for sophoraflavanone G, from 64 to 1024 microg/ml for ampicillin, and from 256 to 1024 microg/ml for oxacillin. The combination of sophoraflavanone G and ampicillin or oxacillin yielded a fractional inhibitory concentration index ranging from 0.188 to 0.375, thereby indicating a principally synergistic effect. The synergistic interaction was verified by time-kill studies using sophoraflavanone G and/or antibiotics. Thirty minutes of treatment with sophoraflavanone G with ampicillin or oxacillin resulted in an increase in the rate of killing in units of CFU/ml to a greater degree than was observed with Sophoraflavanone G alone. These findings indicated that the application of the tested sophoraflavanone G alone or in combination with antibiotics might prove useful in the control and treatment of MRSA infections. PMID- 19288535 TI - The impact of fetal gender on first trimester nuchal translucency and maternal serum free beta-hCG and PAPP-A MoM in normal and trisomy 21 pregnancies. AB - OBJECTIVE: To investigate if fetal sex has an impact on 1st trimester combined screening for aenuploidy. METHODS: We studied the first trimester PAPP-A, free beta-human chorionic gonadatropin (beta-hCG) and nuchal translucency levels in 56,024 normal, singleton pregnancies with known fetal sex at birth. We also examined the distributions in 722 pregnancies with trisomy 21 of known fetal sex. RESULTS: We have found a 14.74% increase in first trimester maternal serum (MS) median free beta-hCG MoM, 6.25% increase of PAPP-A and a 9.41% decrease in delta NT, when the fetus was female. Analysis of data has shown that women carrying a female fetus were 1.084 times more likely to be in the 'at risk' group than those carrying a male fetus. In examining data from 722 pregnancies in which the fetus was affected by trisomy 21, we observed a similar 20.8% increase in free beta-hCG MoM, 5.7% increase in PAPP-A and a 12% decrease in delta NT when the fetus was female. Amongst the trisomy 21 cases, 88.8% of male trisomy 21 cases were detected compared with 91.2% in female cases, this difference was not statistically significant. Correcting for fetal sex redressed the balance in screen-positive rate between the sexes and had a minimal impact on detection rate. CONCLUSION: Correcting for fetal sex may be a worthwhile consideration. A cost-benefit analysis would be required to determine if it is feasible to introduce fetal gender assignment into the routine first trimester scan for the purpose of marker correction and whether this would have any significant impact. PMID- 19288536 TI - Guanidinoacetate methyltransferase (GAMT) deficiency diagnosed by proton NMR spectroscopy of body fluids. AB - In patients with guanidinoacetate methyltransferase (GAMT) deficiency several parameters may point towards the diagnosis of GAMT deficiency. These include the low levels of creatine and creatinine in urine, the high concentration of guanidinoacetic acid (GAA) in urine and the low levels of creatine and creatinine in the cerebrospinal fluid (CSF). In this study, body fluids from 10 GAMT deficient patients were analysed using (1)H NMR spectroscopy. The urine 1D (1)H NMR spectra of all the patients showed a doublet resonance at 3.98 ppm (pH 2.50) derived from GAA present in high concentration. For this compound, a good recovery and good correlation was found between an LC-MS/MS method and (1)H NMR spectroscopy. In CSF NMR spectra of these patients, the singlet resonances of creatine and creatinine (3.05 and 3.13 ppm, respectively) were absent (normally always present in (1)H NMR spectra of CSF). Due to overlap by other resonances, the doublet of GAA could not be observed. Our data demonstrate that (1)H NMR spectroscopy of urine and CSF can be used to diagnose patients with GAMT deficiency. PMID- 19288537 TI - Simultaneous measurement of 13C- and 15N-isotopic enrichments of threonine by mass spectrometry. AB - Under conditions of high isotopic dilution, e.g. in a tracer study, the ability to determine accurately and quantitatively small variations in isotopic enrichments of differently labelled chemical compounds (e.g. (13)C and (15)N in threonine) in a single run by gas chromatography/mass spectrometry (GC/MS) is desirable but remains a technological challenge. Here, we report a new, rapid and simple GC/MS method for simultaneously measuring the isotopic enrichments of doubly labelled threonine ([U(13)C] and (15)N) with isotopic enrichment lower than 1.5 Molar Percent Excess (MPE). The long-term reproducibility measured was around 0.09 MPE for both tracers (throughout a 6 week period). The intra-day repeatability was lower than 0.05 and 0.06 MPE for [U(13)C]-Thr and (15)N-Thr, respectively. To calculate both isotopic enrichments, two modes of calculations were used: one based on work by Rosenblatt et al. in 1992 and the other one using a matrix approach. Both methods gave similar results (ANOVA, P >0.05) with close precision for each mode of calculation. The GC/MS method was then used to investigate the differential utilization of threonine in different organs according to its route of administration in minipigs after administration of both tracers. In plasma samples, the lowest isotopic enrichment measured between two successive time points was at 0.01 and 0.02 MPE for [U(13)C]-Thr and (15)N-Thr, respectively. Moreover, the accuracy of GC/MS (13)C-isotopic enrichment measured was validated by analyzing the same plasma samples by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). Statistical analysis showed that both techniques gave the same results (ANOVA, P >0.05). This new GC/MS method offers the possibility to measure (13)C- and (15)N isotopic enrichments with higher throughput, and using a lower amount of sample, than using GC/C/IRMS. PMID- 19288538 TI - Non-dauer larval dispersal in Caenorhabditis elegans. AB - Species that exploit transient food patches must both effectively utilize such food sources and colonize new patches. The timing and rate of dispersal from existing patches and adaptations that aid dispersal are therefore crucial. Currently, no system exists in which dispersal has been investigated at both the ecological and genetic levels. The extensively studied model nematode Caenorhabditis elegans is potentially such a system. Dispersal between food patches in C. elegans has been found to be related to polymorphism in the npr-1 gene, which regulates the tendency of worms to aggregate on food. Here I show that this non-dauer larval dispersal is affected by various environmental variables and that variation is not fully explained by differences in aggregation behavior. Quantitative trait loci mapping identifies candidate genomic regions, separate to npr-1, which affect variation in dispersal between two isolates. These data suggest that the ecology of C. elegans is more complex than previously thought, but indicate that it is experimentally tractable. PMID- 19288539 TI - False-positive liquid chromatography/tandem mass spectrometric confirmation of sebuthylazine residues using the identification points system according to EU directive 2002/657/EC due to a biogenic insecticide in tarragon. AB - In pesticide residue analysis using liquid chromatography/tandem mass spectrometry (LC/MS/MS) the confirmation of a sebuthylazine finding in a tarragon (Artemisia dranunculus) sample was demonstrated to be false positive. A coeluting interfering matrix compound produced product ions in MS/MS analysis, perfectly corresponding to the multiple reaction monitoring (MRM) of two sebuthylazine transitions. Using the EU directive 2002/657/EC which regulates the confirmation of suspected positive findings would have resulted in a false-positive finding. A third LC/MS/MS transition with a deviant ion ratio and a gas chromatography (GC)/MS/MS analysis revealed the false-positive results. With optimized high resolving ultra-performance liquid chromatography (UPLC) conditions it was possible to separate spiked sebuthylazine from the interfering matrix compound. Using its exact mass and isotope ratios from LC/time-of-flight (TOF) MS measurements, the compound was identified as nepellitorine, a - not surprising - endogenous alkamide in tarragon (Arthemisia dranunculus). False-positive results, especially in heavy matrix samples such as herbs, can be dealt with by further confirmatory analysis, e.g. a third transition, GC analysis if possible or more advantageous by an orthogonal criterion like exact mass. PMID- 19288540 TI - Do schema processes mediate links between parenting and eating pathology? AB - OBJECTIVE: Adverse parenting experiences are commonly linked to eating pathology. A schema-based model of the development and maintenance of eating pathology proposes that one of the potential mediators of the link between parenting and eating pathology might be the development of schema maintenance processes- mechanisms that operate to help the individual avoid intolerable emotions. METHOD: To test this hypothesis, 353 female students and 124 female eating disordered clients were recruited. They completed a measure of perceived parenting experiences as related to schema development (Young Parenting Inventory Revised (YPI-R)), two measures of schema processes (Young Compensatory Inventory; Young-Rygh Avoidance Inventory (YRAI)) and a measure of eating pathology (Eating Disorders Inventory (EDI)). RESULTS: In support of the hypothesis, certain schema processes did mediate the relationship between specific perceptions of parenting and particular forms of eating pathology, although these were different for the clinical and non-clinical samples. DISCUSSION: In those patients where parenting is implicated in the development of eating pathology, treatment might need to target the cognitive processes that can explain this link. PMID- 19288541 TI - An electronic device for accelerating bone formation in tissues surrounding a dental implant. AB - A dental implant is a unique structure which can be used with a noninvasive method because it is inserted into the bone in part and extended extracorporally. This study presents an electronic device that is temporarily connected with the dental implant, and reports its effect on accelerating bone formation in the surrounding tissues in a canine mandibular model. A small sized and low power consumption biphasic electrical current (BEC) stimulator ASIC was developed and the surrounding tissue was exposed to continuous BEC stimulation for 7 days with the parameters of 20 microA/cm(2), 125 micros duration, and 100 pulses/s. After 2 (n = 5) and 5 weeks (n = 5), animals were sacrificed and the specimens were histomorphometrically evaluated. The newly formed bone area (BA) was 1.30 times (3 weeks, P < 0.05) and 1.35 times (5 weeks, P < 0.05) higher in the experimental group compared to the control group, respectively. Bone-implant contact (BIC) in 3-week specimens was 1.62 times (P < 0.05) greater in the experimental group, while there was no statistically significant difference in 5-week specimens. Based on these results showing accelerated bone formation on and around the dental implant, it could be suggested that the latent time for osseointegration in dental implants can be reduced, and the success rate of implants in poor quality bone can be increased by using our device with BEC. PMID- 19288542 TI - Effects of culture medium and protein supplementation on mRNA expression of in vitro produced bovine embryos. AB - Numerous studies have reported aberrant gene expression levels attributed to suboptimal in vitro culture conditions. This study investigated the effects of different culture systems and protein sources on the developmental competence of in vitro production (IVP) embryos measured by cleavage and blastocyst rates, cell number, and relative abundance of POU5F1 (OCT4), nanog, GJA1 (connexin 43), and SLC2A1 (GLUT1) transcripts when compared to in vivo embryos. Experiment 1 compared IVP embryos cultured in either synthetic oviductal fluid (SOFaa) or potassium simplex optimized medium supplemented with amino acids (KSOMaa). Experiment 2 compared the same two culture systems with and without the addition of calf serum (CS). Results from both experiments indicated that despite similar developmental rates, significant differences were observed at the mRNA level. In Experiment 1, OCT4 was the only transcript to have a mean abundance level significantly higher in KSOMaa blastocysts when compared with both SOFaa and in vivo embryos. The same pattern of upregulation of OCT4 mRNA was noted in Experiment 2. There were no significant alterations of the ICM specific transcript nanog in either experiment. In contrast to reports by others, connexin 43 mRNA was not expressed at detectable levels in in vivo embryos analyzed in our studies. Blastocysts cultured in SOFaa with CS or KSOMaa had a significant upregulation of GLUT1 mRNA when compared with other treatments and in vivo embryos. Until differences between IVP and in vivo embryos are minimized, aberrations in IVP will continue to arise. PMID- 19288543 TI - Improved pulse sequences for measurement of small long-range couplings between heteronuclei (29Si-13C) at natural abundance. AB - The gradient pulse sequences for measurement of small long-range couplings between heteronuclei ((29)Si-(13)C) in natural abundance reported to date (INEPT (Si,C)gCOSY and INEPT-(Si,C,Si)HMQC) suffer from significant signal loss when these nuclei ((29)Si, (13)C) are coupled through one-bond couplings to protons. This negative effect can be completely eliminated by using non-gradient versions (INEPT-(Si,C)COSY) or by switching proton decoupling off during gradient pulses (modified INEPT-(Si,C,Si)gHMQC pulse sequence). The beneficial effects of these two approaches on the quality of the spectra are demonstrated here. PMID- 19288544 TI - Immunocontraceptive potential of the Ig-like domain of Izumo. AB - To investigate whether the Ig-like domain of sperm protein Izumo or the other part of the protein could be used as an immunocontraceptive antigen, three partially overlapping cDNA fragments (PA, PB, and PC), together covering entire mouse Izumo, were cloned, expressed, and purified. PB contains the whole Ig-like domain of mouse Izumo. The anti-PB antibody significantly inhibited the fusion of sperm with zona-free mouse eggs with no effect on sperm motility, while anti-PA and anti-PC antibodies virtually had no effect on sperm-egg fusion at the same concentration. Furthermore, in the presence of anti-PB antibody, the anti-sperm reactivity could be competitively inhibited by recombinant PB protein. The PB specific antibody staining was restricted to the acrosome region in acrosome reacted mouse spermatozoa by indirect immunofluorescence. Active immunization with the PB antigen sharply raised the antibody titers in mouse that were enough to cause a significant reduction in fertility compared to the PA and PC immunized groups. In conclusion, our data indicate that the Ig-like domain of Izumo plays an important role in the fertilization process, as verified by the dose-dependent reduction in fertilization rates in mouse IVF trials and mouse mating assay. These results indicate that the Ig-like domain of Izumo might be a new candidate for the development of a contraceptive vaccine. PMID- 19288545 TI - A rapid flow cytometric screening test for X-linked lymphoproliferative disease due to XIAP deficiency. AB - BACKGROUND: Deficiency of X-linked inhibitor of apoptosis (XIAP), caused by BIRC4 gene mutations, is the second known cause of X-linked lymphoproliferative disease (XLP), a rare primary immunodeficiency that often presents with life-threatening hemophagocytic lymphohistiocytosis (HLH). Rapid diagnosis of the known genetic causes of HLH, including XIAP deficiency, facilitates the initiation of life saving treatment and preparation for allogeneic hematopoietic cell transplantation (HCT). Until now, a rapid screening test for XIAP deficiency has not been available. METHODS: To develop a flow cytometric screening test for XIAP deficiency, we first used lymphoblastic cell lines generated from controls and patients with BIRC4 mutations to identify two commercially available antibodies specific for native intracellular XIAP. Next, we used these antibodies to study control whole blood leukocyte XIAP expression. We then studied XIAP expression in leukocytes from patients with XLP due to BIRC4 mutations, maternal carriers, and patients following HCT. RESULTS: XIAP was expressed by the majority of all whole blood nucleated cells in normal controls. In contrast, XIAP was absent or decreased in all lymphocyte subsets, monocytes and granulocytes from four unrelated patients with XLP due to BIRC4 mutations. Bimodal distribution of XIAP expression was evident in two maternal carriers, with significant skewing toward cells expressing normal XIAP. Bimodal distribution was also observed in a patient following HCT. CONCLUSIONS: Flow cytometric analysis of intracellular XIAP provides a rapid screening test for XLP due to XIAP deficiency. It also allows carrier detection and can be used to monitor donor versus recipient reconstitution following HCT. PMID- 19288546 TI - Complete assignments of (1)H and (13)C NMR data for two new sesquiterpenes from Cyperus rotundus L. AB - Two new sesquiterpenes, epi-guaidiol A (1) and sugebiol (3), together with four known sesquiterpenes, guaidiol A(2), sugetriol triacetate (4), cyperenoic acid (5), and cyperotundone (6) were isolated from the rhizomes of Cyperus rotundus L. Their structures were identified by MS and NMR experiments, and the complete assignments of (1)H and (13)C NMR data for two new sesquiterpenes were obtained by the aid of two-dimensional (2D) NMR techniques, including HSQC, HMBC, (1)H (1)HCOSY and nuclear overhauser enhancement spectroscopy(NOESY). PMID- 19288547 TI - Phagocytosis and postphagocytic reaction of cord blood and adult blood monocyte after infection with green fluorescent protein-labeled Escherichia coli and group B Streptococci. AB - BACKGROUND: Neonatal sepsis is characterized by an excessive inflammatory response induced by immune cells (monocytes). We investigated the initial stage of monocyte-pathogen interaction, i.e. bacterial ingestion and degradation at the single-cell level, by comparing a new flow cytometric procedure with culture methods. We also examined the hypothesis that, in terms of phagocytosis-induced cell death (PICD), phenotype, or cytokine production, cord blood monocytes (CBMO) differ from monocytes derived from adults (peripheral blood monocytes, PBMO). METHODS: Phagocytosis and intracellular degradation were assessed by means of flow cytometry and bacterial cultures of green fluorescent protein-labeled group B Streptococci (GBS) and Escherichia coli. The production of reactive oxygen species (ROS) was measured through luminol-enhanced chemiluminescence. Apoptosis, phenotype, and cytokine production were assessed through flow cytometry. RESULTS: Flow cytometry and bacterial cultures showed no difference between phagocytosis and degradation of GBS and E. coli by PBMO and CBMO. A high correlation between both methods was observed. No difference in ROS production was evident. In comparison with PBMO, CBMO apoptosis was lower after exposure to GBS and E. coli. Similarities were found between nonapoptotic monocytes and pro-inflammatory monocytes. CONCLUSIONS: PICD is lower in CBMO during the early stages of monocyte pathogen interaction. Our results emphasize that monocyte apoptosis has a potential role in tailoring the immune response in neonatal sepsis. PMID- 19288548 TI - Revised delta34S reference values for IAEA sulfur isotope reference materials S-2 and S-3. AB - Revised delta(34)S reference values with associated expanded uncertainties (95% confidence interval (C.I.)) are presented for the sulfur isotope reference materials IAEA-S-2 (22.62 +/- 0.16 per thousand) and IAEA-S-3 (-32.49 +/- 0.16 per thousand). These revised values are determined using two relative-difference measurement techniques, gas source isotope ratio mass spectrometry (GIRMS) and double-spike multi-collector thermal ionization mass spectrometry (MC-TIMS). Gas analyses have traditionally been considered the most robust for relative isotopic difference measurements of sulfur. The double-spike MC-TIMS technique provides an independent method for value-assignment validation and produces revised values that are both unbiased and more precise than previous value assignments. Unbiased delta(34)S values are required to anchor the positive and negative end members of the sulfur delta (delta) scale because they are the basis for reporting both delta(34)S values and the derived mass-independent Delta(33)S and Delta(36)S values. PMID- 19288549 TI - Chiral NMR discrimination of the diastereoisomeric salts of the H3-antagonist 2 [3-(1H-imidazol-4-ylmethyl)piperidin-1-yl]-1H-benzimidazole. AB - Diastereomeric salts with optically pure (S)-alpha-methoxy-alpha (trifluoromethyl)phenylacetic acid (MTPA) were used to discriminate the enantiomers of the chiral H(3)-antagonist 2-[3-(1H-imidazol-4-ylmethyl)piperidin 1-yl]-1H-benzimidazole. Chemical-shift differences (Delta delta) in NMR spectra strongly depend on solvent and stoichiometric ratio. The better observable differentiation occurred for the proton at the 2-position of the imidazole ring. PMID- 19288550 TI - Postnatal testicular regression mircopenis and microcephaly: conformation of a new syndrome? AB - Testicular regression may develop at various phases of the intrauterine period and the clinical findings vary depending on the timing of the intrauterine phase. On the other hand, postnatal regression of the testicles is rare and few cases have been reported in the literature. Our patient presented with undescended testicles, micropenis, flat scrotum and microcephaly. The patient's levels of basal luteinizing hormone (LH) were low/normal and follicle stimulating hormone (FSH) was high. No response was obtained in the human chorionic gonadotropin (HCG) stimulation test. The levels of inhibin B and anti Mullerian hormone (AMH) were found to be low. Penile growth response to intramuscular testosterone injections was 2.5 cm. Testicles were visualized bilaterally in inguinal canal by ultrasound examination initially but by 2 years of age no testicular tissue was observed during inguinal exploration. In conclusion we recognized postnatal testicular regression in our patient that had started in the intrauterine period and persisted into infancy. The genital system anomalies, microcephaly and motor retardation in our patient confirm the hypothesis of Parisi et al. of a novel condition of postnatal regression and micropenis. PMID- 19288551 TI - Two distinctive classic genetic syndromes, 22q11.2 deletion syndrome and Angelman syndrome, occurring within the same family. AB - We document a sib pair born to a mother with a reciprocal translocation, t(15;22)(q13;q11.2): the daughter had the Angelman syndrome phenotype associated with a maternally derived 15q deletion, and the son had a phenotype associated with a 22q deletion. Adjacent two-type segregation during gametogenesis in the mother can account for the unbalanced karyotypes of the siblings. From a tetravalent chromatid formed by normal chromosome 15, derivative chromosome 15, normal chromosome 22, and derivative chromosome 22, the daughter inherited chromosome 22 and derivative chromosome 22 and the son inherited chromosome 15 and derivative chromosome 15. The family is unique in that two distinctive genetic syndromes, 22q11.2 deletion syndrome and Angelman syndrome, occurred within the same family. The family is also elucidative from an educational standpoint in that major concepts of non-Mendelian inheritance (microdeletion, genomic imprinting, and reciprocal translocation) need to be considered to appreciate the inheritance pattern. Furthermore, the family illustrates the importance of cryptic rearrangements at the most proximal end of acrocentric chromosomes in the evaluation of siblings with multiple congenital anomaly-mental retardation phenotypes that are dissimilar among affected siblings. The situation is analogous to parental balanced translocation between the most "distal" segments of a chromosome, that is, the subtelomere region, a recently appreciated cause of familial recurrence of multiple congenital anomaly-mental retardation phenotype with a normal G-banding karyotype. We suggest that cryptic rearrangements at the most proximal end, analogous to those at the most distal end, should be considered as an appreciable cause of recurrent multiple congenital anomaly-mental retardation phenotype. PMID- 19288552 TI - Multiple joint dislocations: an additional skeletal finding in Lowry-Wood syndrome? AB - We report on the case of a 17-year-old boy with clinical features compatible with Lowry-Wood syndrome: microcephaly, short stature, multiple epiphyseal dysplasia, tapetoretinal degeneration, and mental retardation. Bilateral restricted elbow extension, knock knees and hip dislocation were also present. Radiographs showed evidence of radial dislocation due to the absence of the radial heads, lateral dislocation of both patellae, multiple epiphyseal dysplasia that was more severe at the proximal femoral epiphyses, and dislocation of both hips with severe hip dysplasia. The patient developed a behavioral disorder at age 15. Conventional karyotyping was normal (46,XY). Molecular karyotyping, performed through array based competitive genomic hybridization, showed copy number variants that were probably benign. We suggest that multiple joint dislocations, including the patellae, may be a sign of Lowry-Wood syndrome. PMID- 19288553 TI - The feeding ecology and activity budget of proboscis monkeys. AB - A group of proboscis monkeys (Nasalis larvatus) consisting of an alpha-male, six adult females, and several immatures was observed from May 2005-2006. We collected over 1,968 hr of focal data on the adult male and 1,539 hr of focal data on the six females in a forest along the Menanggul River, Sabah, Malaysia. Availability and seasonal changes in plant species consumed by the focal monkeys were determined by vegetation surveys carried out across an area of 2.15 ha along 200-500 m trails in riverine forest. A total of 188 plant species were consumed by the focal monkeys. The activity budget of members of our study group was 76.5% resting, 19.5% feeding, and 3.5% moving. Young leaves (65.9%) and fruits (25.9%) accounted for the majority of feeding time. Over 90% of fruit feeding involved the consumption of unripe fruits and in the majority of case both the fruit flesh and seeds were eaten. Although fruit eating was rare in some months, during other times of the year time fruit feeding exceeded the time devoted to young leaves. We found that monthly fruit availability was positively related to monthly fruit eating and feeding activity, and seasonal fluctuations in dietary diversity were significantly affected by fruit eating. These results suggest that fruit availability and fruit-eating behaviors are key factors that influence the activity budget of proboscis monkeys. Earlier assumptions that colobine monkeys are obligate folivores do not apply well to proboscis monkeys and certain other colobines. Our findings may help contribute to a better understanding of the dietary adaptations and feeding ecology of Asian colobines. PMID- 19288554 TI - Polyhydramnios, fetal overgrowth, and macrocephaly: prenatal ultrasound findings of Costello syndrome. AB - Costello syndrome is a multiple congenital anomaly syndrome consisting of dysmorphic facies, cutis laxa, short stature, developmental delay, and mental retardation. Complications include failure to thrive, hypertrophic cardiomyopathy with arrhythmias, and benign and malignant tumors. This report describes a new case of Costello syndrome in a preterm infant born at 27 weeks gestation and diagnosed with Costello syndrome at 7 weeks of life who died at 6 months of age due to cardiac and pulmonary complications. In addition, data were compiled from parent surveys including growth parameters on 16 infants who were subsequently diagnosed with Costello syndrome and had mutation confirmation. The most common prenatal findings in the literature and in this cohort were polyhydramnios and fetal overgrowth with relative macrocephaly. Based on this study, ultrasound identification of polyhydramnios in the context of prenatal overgrowth, especially with relative macrocephaly, needs to raise the possibility of a diagnosis of Costello syndrome in the fetus because of the life-threatening cardiac complications that may occur early in the newborn period. PMID- 19288555 TI - Homozygous structural rearrangement 16p13: a mechanism of tumorigenesis in sporadic renal angiomyolipoma? PMID- 19288556 TI - Who is driving continuing medical education for family medicine? AB - INTRODUCTION: Considerable time and money are invested in continuing medical education (CME) for family physicians (FPs) but the effectiveness is uncertain. The participation of FPs as coordinators and teachers is not well known. The goal of this project was to describe the role of FPs in organizing and teaching CME events that are accredited for FPs. METHODS: Information about accredited CME events occurring in Alberta and Nova Scotia was requested from universities, provincial chapters of the College of Family Physicians of Canada, and pharmaceutical companies. Location, coordinating site, organizing committee members, teaching faculty, and format were recorded from each event. The number and proportion of FPs involved in both organizing and teaching CME events accredited for FPs were calculated and compared. RESULTS: A total of 314 CME events were collected, comprising a total of 1,472 hours of CME. From the CME events collected, there were 1,730 organizing committee members and 1,647 teachers. FPs constitute 59% of the organizing committees and 17% of the teachers. Significant differences in the numbers of FP planners and teachers were related to organizing group, format, location, and expected audience composition. DISCUSSION: The accreditation requirement for FPs on organizing committees likely helps preserve a reasonable proportion of FP organizers but not teachers in FP CME. The proportions of true FP planners and teachers may actually be lower than planning documents indicate. Low level of family physician teachers in CME may be due to FPs' not selecting FP teachers, the FP teaching pool's being inadequate, or the organizing committee's being unaware of FPs who are knowledgeable in particular areas. PMID- 19288557 TI - Examining the value of commercially supported CME. AB - Commercial support of continuing medical education/continuing professional development (CME/CPD) is a fact of life currently, though under attack from several sources. Does it have a positive or negative value to industry, to physicians, to society, and to CME professionals? There is sufficient evidence to support positive value to industry. There is insufficient evidence to support positive or negative value to physicians or society. There is reason to continue commercial support while broadening the base of support beyond the pharmaceutical industry, not only to avoid perception of bias but also to address CME/CPD needs that do not fit in therapeutic categories. CME professionalism does not depend on commercial support. PMID- 19288558 TI - A view from industry: the foundations of future commercial support and a call for action. AB - Based on a model of industry support that aligns with the emerging performance improvement approach to continuing medical education (CME), a call to action is made for the transformation of commercial support. Today, commercial support, like the CME profession, is linked largely to educational activities of far less value to patients than the model now emerging to address professional practice gaps. The new model could help ameliorate lingering concerns about the presence of commercial support, but it also will require fundamental changes in perspective and procedures by both industry and CME providers. PMID- 19288559 TI - Audience assessment of bias in continuing medical education programs. PMID- 19288560 TI - Back to school for the teachers: improving teaching of physical diagnosis. PMID- 19288561 TI - Obesity syndrome, MOMES caused by deletion-duplication (4q35.1 del and 5p14.3 dup). PMID- 19288562 TI - Achieving desired results and improved outcomes: integrating planning and assessment throughout learning activities. AB - Most physicians believe that to provide the best possible care to their patients, they must commit to continuous learning. For the most part, it appears the learning activities currently available to physicians do not provide opportunities for meaningful continuous learning. At the same time there have been increasing concerns about the quality of health care, and a variety of groups within organized medicine have proposed approaches to address issues of physician competence and performance. The authors question whether CME will be accepted as a full partner in these new approaches if providers continue to use current approaches to planning and assessing CME. A conceptual model is proposed for planning and assessing continuous learning for physicians that the authors believe will help CME planners address issues of physician competence, physician performance, and patient health status. PMID- 19288563 TI - Improved cardiovascular prevention using best CME practices: a randomized trial. AB - INTRODUCTION: It was hypothesized that after a continuing medical education (CME) event, practice enablers and reinforcers addressing main clinical barriers to preventive care would be more effective in improving general practitioners' (GPs) adherence to cardiovascular guidelines than a CME event only. METHODS: A cluster randomized trial was conducted on a convenience sample of 122 GPs who were randomly assigned to either CME only (control group) or CME with practice enablers and reinforcers (PER group). In the PER group, nurses visited GPs' offices once a month to implement the clinical intervention on patients > or = 55 years old with a scheduled visit in the month following the nurse visit: (1) screening medical records for potentially undermanaged high-risk patients; (2) prompting physicians to reassess preventive care in these patients; (3) enclosing a checklist reporting most recent information relevant to guidelines' implementation; and (4) enclosing a summary of experts' recommendations in the form of a follow-up and treatment algorithm. RESULTS: A retrospective chart audit of 2344 consenting patients, potentially undermanaged at baseline, demonstrated that the PER intervention following CME significantly improved adherence to guidelines compared to CME alone (OR: 1.78, 95% CI: 1.32-2.41). DISCUSSION: The intervention was designed for self-implementation in primary care practices that have their own nursing staff. PER GPs were highly satisfied with the intervention; the majority said that they would implement it in their practice if someone trained their nurse, thus suggesting support for development of a multiprofessional CME program to disseminate this clinical approach to primary care practice groups. PMID- 19288564 TI - Knowledge to action: scholarship for faculty and staff. AB - INTRODUCTION: This study was undertaken to evaluate the influence of a continuing education course in facilitating the development and implementation of educational projects of course participants. METHODS: This is a case study evaluating a full-year course that consisted of 11 monthly seminars, each 4 hours in length, including practice in a computer laboratory. The class size was limited to 12 participants. Needs-assessment surveys at the beginning of the course, student evaluations, and midterm and final progress reports were analyzed. RESULTS: Seven staff physicians, 3 clinical fellows, a nurse educator, and a research assistant enrolled in the course. Initial needs-assessment surveys indicated that most people had adequate computer skills-11 (90%)-but only 2 (17%) were able to type well, 11 (90%) had no statistical knowledge, and 10 (83%) had limited literature-searching skills. The mean score on speaker evaluations for lectures was 4.5 on a scoring scheme of 1-5 where 1 was poor and 5 was outstanding. Ten participants (83%) had a complete proposal for an educational project written by midterm. Nine participants applied for external grants and 2 of them received external funding for their projects. Five participants (42%) completed a publishable educational project by the end of the 11-month course, and submitted it for presentation at scientific meetings. DISCUSSION: Like many adults, health care professionals experience limited time for involvement in formal education. This study shows that a limited-time-commitment course could facilitate health care professionals to develop and successfully implement educational projects translating ideas into action. PMID- 19288565 TI - Internet point of care learning at a community hospital. AB - INTRODUCTION: Internet point of care (PoC) learning is a relatively new method for obtaining continuing medical education credits. Few data are available to describe physician utilization of this CME activity. METHODS: We describe the Internet point of care system we developed at a medium-sized community hospital and report on its first year of utilization. We reviewed all completed Internet PoC report forms for 1 year to determine what percentage of our medical staff participated. Using a Likert-type scale, physicians were asked to rate the effectiveness of this method as a means for obtaining CME credit and for improving patient care. RESULTS: From a medical staff of 355, 7 physicians (2%) completed an Internet PoC form in the first year of use. The number of completed forms ranged from 1 to 18 for each individual who used the system. On a 5-point Likert scale, physicians rated this method as follows: good answer found to question = 4.4, knowledge base was improved = 4.0, patient benefited from the information = 4.0, and good way to learn new information = 4.2. DISCUSSION: A small number of physicians utilized this method for obtaining CME credit, but some individuals really embraced it. Those who used it liked it as a way to obtain CME and many felt strongly that it improved patient care. Other hospitals are encouraged to develop an Internet PoC reporting system for their medical staff. PMID- 19288566 TI - Electronic continuing education in the health professions: an update on evidence from RCTs. AB - INTRODUCTION: Demonstrating the effectiveness of the rapidly expanding field of electronic continuing education (e-CE) has important implications for CE in the health professions. This study provides an update on evidence from randomized controlled trials (RCTs) assessing the effectiveness of e-CE in the health professions. METHODS: A literature search of RCTs was performed in MEDLINE, EMBASE, and CINAHL from 2004 to 2007. Papers were reviewed separately by 2 of the authors and results were categorized and reviewed according to study comparisons. RESULTS: Fifteen studies met our inclusion criteria. Six compared e-CE to no intervention or placebo. Of these 6 studies, 4 showed a statistically significant advantage of the e-CE intervention and 2 showed no significant effect. Two studies compared e-CE to a lecture. Of these, 1 showed an advantage of e-CE and 1 showed no difference. Two studies compared e-CE to a small-group interactive intervention. In both studies, the e-CE group outperformed the control. Two studies compared a multicomponent e-CE intervention to one based on flat text, and both showed the multicomponent intervention to be more effective. Two of the 15 studies demonstrated a statistically significant effect on practice patterns. Positive effects of e-CE on knowledge were shown to persist for up to 12 months and effects on practice up to 5 months. DISCUSSION: Overall, these studies suggest that multicomponent e-CE interventions can be effective in changing health professionals' practice patterns, and improve their knowledge. E-CE interventions based purely on flat text appear to be of limited effectiveness in changing either knowledge or practice. These results support the use of multicomponent e-CE as a method of CE delivery. PMID- 19288567 TI - Community pharmacists' preferences for continuing education delivery in Australia. AB - INTRODUCTION: New technologies such as the Internet offer an increasing number of options for the delivery of continuing education (CE) to community pharmacists. Many of these options are being utilized to overcome access- and cost-related problems. This paper identifies learning preferences of Australian community pharmacists for CE and identifies issues with the integration of these into contemporary models of CE delivery. METHODS: Four focus group teleconferences were conducted with practicing community pharmacists (n = 15) using a semistructured format and asking generally about their CE and continuing professional development (CPD) experiences. RESULTS: Pharmacists reported preferences for CE that were very closely aligned to the principles of adult learning. There was a strong preference for interactive and multidisciplinary CE. Engaging in CPD was seen as valuable in promoting reflective learning. DISCUSSION: These results suggest that pharmacists have a strong preference for CE that is based on adult learning principles. Professional organizations should take note of this and ensure that new CE formats do not compromise the ability of pharmacists to engage in interactive, multidisciplinary, and problem-based CE. Equally, the role of attendance-based CE in maintaining peer networks should not be overlooked. PMID- 19288568 TI - Stage migration after minor changes in histologic estimation of tumor burden in sentinel lymph nodes: the protocol trap. AB - BACKGROUND: Melanoma metastasis size estimates are of prognostic significance for groups of patients, but to the authors' knowledge, measurement consensus does not exist. METHODS: Maximum metastasis diameter, maximum centripetal tumor depth, microanatomic location of metastases, and complete metastasis volume were measured in 156 positive sentinel lymph nodes (SLNs) from 99 melanoma patients. RESULTS: The number of SLN-positive patients was increased by up to 41% using complete step-sectioning compared with less extensive protocols. Assessing maximum metastasis diameters, up to 27% of patients positive by the less extensive protocols went from 1 metastasis diameter group to a larger one when complete step-sectioning was performed. No patients were down-staged. Apparently minor protocol changes (eg, adding an extra step) led to substantial changes in maximum metastasis diameter. Similar protocol-dependent results were noted measuring the maximum centripetal tumor depth and the microanatomical location of metastases. By using semiquantitative tumor burden estimates, stage migration was always unidirectional (ie, moving from a lower to higher stage). Stereologic tumor burden estimates in step-sectioned SLNs also varied according to the number of step sections assessed, but could increase, decrease, or remain constant, so that stage migration was multidirectional. CONCLUSIONS: Adding extra steps to pathology protocols when assessing semiquantitative parameters leads to unidirectional stage migration ("the protocol trap"). This systematical bias makes it difficult to base treatment decisions on semiquantitative metastasis size estimates. Although based on metastatic melanoma, the principles described herein will apply when measuring nodal tumor burden in other metastasizing cancers, including breast carcinoma. PMID- 19288569 TI - Patterns of treatment for early stage breast cancers at the M. D. Anderson Cancer Center from 1997 to 2004. AB - BACKGROUND: The objectives of this study were to examine the patterns of use for adjuvant therapy and the changes in surgical practice for patients with early stage breast cancer and to describe how recent large clinical trial results impacted the patterns of care at The University of Texas M. D. Anderson Cancer Center (MDACC). METHODS: The study included 5486 women who were diagnosed with stage I through IIIA breast cancer between 1997 and 2004 and received their treatment at MDACC. A chi-square trend test and multivariate logistic regression model were used to assess changes in treatment patterns over time. RESULTS: Among lymph node-positive patients, the use of anthracycline plus taxane chemotherapy increased from 17% in 1997 to 81% in 2004 (P < .001). Meanwhile, the use of anthracyclines without taxanes dropped from 76% to 20% (P < .001) between 1997 and 2000. For postmenopausal patients who received endocrine therapy, the use of tamoxifen was replaced increasingly by the use of aromatase inhibitors (from 100% on tamoxifen in 1997 to 14% in 2004; P < .001). The percentage of women who underwent initial sentinel lymph node biopsy increased significantly during the period from 1997 to 2004 (from 1.8% to 69.7%, respectively, among patients who underwent mastectomy; and from 18.1% to 87.1%, respectively, among patients who underwent breast-conserving surgery; P < .001). CONCLUSIONS: The results from this study suggested that key findings from adjuvant therapy and surgical procedures from large clinical trials often prompt immediate changes in the patient care practices of research hospitals like MDACC. PMID- 19288570 TI - A novel hypothesis on the sensitivity of the fecal occult blood test: Results of a joint analysis of 3 randomized controlled trials. AB - BACKGROUND: Estimates of the fecal occult blood test (FOBT) (Hemoccult II) sensitivity differed widely between screening trials and led to divergent conclusions on the effects of FOBT screening. We used microsimulation modeling to estimate a preclinical colorectal cancer (CRC) duration and sensitivity for unrehydrated FOBT from the data of 3 randomized controlled trials of Minnesota, Nottingham, and Funen. In addition to 2 usual hypotheses on the sensitivity of FOBT, we tested a novel hypothesis where sensitivity is linked to the stage of clinical diagnosis in the situation without screening. METHODS: We used the MISCAN-Colon microsimulation model to estimate sensitivity and duration, accounting for differences between the trials in demography, background incidence, and trial design. We tested 3 hypotheses for FOBT sensitivity: sensitivity is the same for all preclinical CRC stages, sensitivity increases with each stage, and sensitivity is higher for the stage in which the cancer would have been diagnosed in the absence of screening than for earlier stages. Goodness-of-fit was evaluated by comparing expected and observed rates of screen detected and interval CRC. RESULTS: The hypothesis with a higher sensitivity in the stage of clinical diagnosis gave the best fit. Under this hypothesis, sensitivity of FOBT was 51% in the stage of clinical diagnosis and 19% in earlier stages. The average duration of preclinical CRC was estimated at 6.7 years. CONCLUSIONS: Our analysis corroborated a long duration of preclinical CRC, with FOBT most sensitive in the stage of clinical diagnosis. PMID- 19288571 TI - Efficacy of repeat isolated limb infusion with melphalan and actinomycin D for recurrent melanoma. AB - BACKGROUND: : Isolated limb infusion (ILI) is an effective and minimally invasive treatment option for delivering regional chemotherapy in patients with metastatic melanoma confined to a limb. Recurrent or progressive disease after an ILI, however, presents a challenge for further treatment. The value of repeat ILI in this situation has not been well documented. METHODS: : Forty-eight patients were identified who had been treated with a repeat ILI. In all patients, a cytotoxic combination of melphalan and actinomycin D was used. RESULTS: : The median time between the 2 procedures was 11 months. The complete response (CR) rate after repeat ILI was 23%, compared with 31% after the initial ILI (P = .36). The overall response was 83%, compared with 75% after the first procedure (P = .32). The median duration of response was 11 months (10 months for patients with CR; P = .80), and median survival was 38 months. In those patients achieving a CR, the median survival was 68 months (P = .003). Toxicity after repeat ILI was increased, with 20 patients experiencing Wieberdink grade III limb toxicity (considerable erythema and edema with blistering) and 5 patients experiencing grade IV toxicity (threatened or actual compartment syndrome), whereas after the initial ILI these toxicity grades occurred in 14 patients and 1 patient, respectively (P = .03). No patient experienced grade V toxicity (requiring amputation). CONCLUSIONS: : Repeat ILI is an attractive treatment option to achieve limb salvage in patients with inoperable recurrent or progressive melanoma after a previous ILI. It can be associated with significant short-term regional toxicity, but is well tolerated by most patients, with satisfactory response rates. Cancer 2009. (c) 2009 American Cancer Society. PMID- 19288572 TI - Neoadjuvant chemotherapy/gefitinib followed by concurrent chemotherapy/radiation therapy/gefitinib for patients with locally advanced squamous carcinoma of the head and neck. AB - BACKGROUND: The authors evaluated the feasibility, toxicity, and efficacy of gefitinib added to first-line combined-modality therapy for patients with locally advanced squamous carcinoma of the head and neck. METHODS: Patients with biopsy proven locally advanced squamous carcinoma of the head and neck who had low expected cure rates with local treatment modalities alone were eligible for this treatment. All patients received a 6-week induction course of docetaxel, carboplatin, infusional 5-fluorouracil, and gefitinib (250 mg daily). Gefitinib was continued while patients received concurrent weekly docetaxel and radiation therapy. After the completion of radiation therapy, gefitinib was continued until patients developed disease progression or for a maximum of 24 months. RESULTS: Sixty-two patients (53% with stage IV disease) received protocol treatment, and 50 patients (81%) were able to complete the regimen. The addition of gefitinib increased the incidence of grade 3/4 mucositis (27%) and diarrhea (16%) during induction therapy but did not appear to add substantially to toxicity during concurrent chemoradiation. The estimated 3-year progression-free and overall survival rates for the entire group were 41% and 54%, respectively. CONCLUSIONS: The addition of gefitinib was associated with a moderate increase in toxicity with this combined modality regimen, particularly during induction therapy. Although this regimen was efficacious, the survival results overlap with results reported with chemoradiation alone. The role of epidermal growth factor receptor inhibitors in first-line, combined-modality therapy for patients with head and neck cancer remains undefined. PMID- 19288573 TI - Prognostic models: from the fates to the future. PMID- 19288574 TI - Genistein enhances the effect of epidermal growth factor receptor tyrosine kinase inhibitors and inhibits nuclear factor kappa B in nonsmall cell lung cancer cell lines. AB - BACKGROUND: Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs) have shown modest clinical benefit in patients with relapsed nonsmall cell lung cancer (NSCLC). Down-regulation of Akt appears to correlate with the antitumor activity of EGFR-TKIs. Akt activates nuclear factor kappa B (NF kappaB), which transcribes genes important for cell survival, invasion, and metastasis. The authors hypothesized that genistein, through the inhibition of NF kappaB, could enhance the activity of EGFR-TKIs in NSCLCs. METHODS: Three NSCLC cell lines with various EGFR mutation status and sensitivities to EGFR-TKIs were selected: H3255 (L858R), H1650 (del E746-A750), and H1781 (wild-type EGFR). Cells were treated with erlotinib, gefitinib, genistein, or the combination of each of the EGFR-TKIs with genistein. Cell survival and apoptosis were assessed, and expression levels of EGFR, pAkt, cyclooxygenase-2 (COX-2), E-cadherin, prostaglandin E(2) (PGE(2)), and NF-kappaB were measured. RESULTS: Both EGFR-TKIs demonstrated growth inhibition and apoptosis in each of the cell lines, but H1650 and H1781 were much less sensitive. Genistein demonstrated some antitumor activity in all cell lines, but enhanced growth inhibition and apoptosis when combined with the EGFR-TKIs in each of the cell lines. Both combinations down regulated NF-kappaB significantly more than either agent alone in H3255. In addition, the combinations reduced the expression of EGFR, pAkt, COX-2, and PGE(2,) consistent with inactivation of NF-kappaB. CONCLUSIONS: The authors concluded that genistein enhances the antitumor effects of EGFR-TKIs in 3 separate NSCLC cell lines. This enhanced activity is in part because of greater reduction in the DNA-binding activity of NF-kappaB when EGFR-TKIs were combined with genistein. PMID- 19288575 TI - Continuing medical education and professional revalidation in Europe: five case examples. AB - INTRODUCTION: Since reliable information is scarce to describe continuing medical education (CME) and revalidation in Europe, we carried out a survey in 5 selected countries (France, Germany, Italy, Spain, and the United Kingdom). METHODS: A tested questionnaire was sent to 2 experts per country (except in Germany), during August-September 2004. RESULTS: In the analyzed countries medical societies, medical associations, and employers are the main CME providers. Pharmaceutical industry sponsorship accounts on the average for more than 50% of the CME financing in those countries. In all 5 countries, CME accreditation systems exist; the national health authorities and medical associations are mainly responsible for them. In France, Italy, and Germany CME is mandatory; in Spain and the United Kingdom it is voluntary. CME credits/points are mainly used for professional career purposes. Revalidation systems have not been introduced in any of these countries, although in the United Kingdom it is being introduced as part of a relicensing process. DISCUSSION: Recommendations for the implementation of a European system of CME/CPD harmonization are made by the authors. PMID- 19288576 TI - Patient selection for hemiablative focal therapy of prostate cancer: variables predictive of tumor unilaterality based upon radical prostatectomy. AB - BACKGROUND: The application of focal therapy for low-risk prostate cancer (PCa) depended on appropriate patient selection. No definitive criteria existed to characterize patients who may potentially benefit from an organ-sparing approach. We evaluated pretreatment clinical parameters that may predict unilateral PCa amenable to hemigland thermoablation. METHODS: In total, 538 patients with complete data from the Duke Prostate Center (DPC) Outcomes database with low- to low-intermediate-risk PCa (prostate-specific antigen<10 ng/mL, biopsy Gleason score < or =7, and clinical stage T1c-T2b) treated with radical prostatectomy (RP) were included in the dataset. Patients underwent diagnostic prostate biopsy (PBx) at Duke or community hospitals from 1996 to 2006. Clinical and biopsy parameters were assessed as to the ability to predict PCa unilaterality verified by RP pathology. RESULTS: The strongest predictor of pathologic unilaterality was PBx unilaterality. The sensitivity and specificity for biopsy unilaterality to predict pathologic unilaterality was 88.4% and 34%, with a positive predictive value of 28% and a negative predictive value of 91%. PBx unilaterality (odds ratio [OR] = 3.88; 95% confidence interval [CI], 2.14-7.05; P < .0005) and negative family history of PCa (OR = 1.83; 95% CI, 1.09-3.05; P = .21) was associated with a higher probability of unilateral disease by multivariate regression. CONCLUSIONS: Two pretreatment clinical variables were significantly predictive of unilateral PCa: negative family history of PCa and PBx unilaterality. These variables may be used to select men with low- to low moderate-risk PCa for hemiablation. Further work is necessary to decrease the false-negative and false-positive rates associated with PBx to improve predictability for PCa laterality. PMID- 19288577 TI - In vivo [(123)I]CNS-1261 binding to D-serine-activated and MK801-blocked NMDA receptors: A storage phosphor imaging study in rats. AB - Disturbances of activity of the glutamatergic neurotransmitter system in the brain are present in many neuropsychiatric disorders. The N-methyl-D-aspartate (NMDA) receptor is the most abundant receptor of the glutamatergic system. In the neurodegenerative events of Alzheimer's disease, excessive activation of NMDA receptors may contribute to neuronal death. Inhibition of NMDA receptor activation may have neuroprotective effects and (semi)quantitative imaging of the activated system may help in the selection of patients for such inhibition therapies. In this study we evaluated [(123)I]CNS-1261 binding in the rat brain. This radiotracer binds in vivo to the MK801 binding site of activated NMDA receptors. To determine the optimal time point for ex vivo assessments after bolus injection [(123)I]CNS-1261 binding in rats, we performed a time course biodistribution study using dissection techniques. [(123)I]CNS-1261 binding was also studied in the rat brain using autoradiography by means of storage phosphor imaging, with prior facilitation of NMDA receptor activation by injection of the potent coagonist D-serine and after blocking of the NMDA receptor binding site by MK801 injection in D-serine pretreated rats. Measurements of [(123)I]CNS-1261 uptake matched the distribution of similar tracers for the MK801 binding site of the NMDA receptor and revealed an optimal time point of 2 h post injection for the assessment of tracer distribution in the rat brain. The blocking experiments indicated specific binding of [(123)I]CNS-1261 to NMDA receptors but also a considerable amount of nonspecific binding. Facilitation of NMDA receptor activation by D-serine did not result in an enhancement of binding of the radiotracer in the NMDA receptor-rich rat hippocampus compared to the untreated group, as measured by autoradiography. In conclusion, our study has shown that [(123)I]CNS-1261 binding is influenced by NMDA receptor availability. However, high nonspecific binding limits quantification and small changes in receptor availability are unlikely to be detected. PMID- 19288578 TI - Reported childhood abuse is associated with low serotonin transporter binding in vivo in major depressive disorder. AB - BACKGROUND: Physical or psychological adversity in childhood is associated with a higher risk for depression in adulthood, and with persistent serotonergic abnormalities in humans and in animal models. We hypothesized that reported childhood abuse would be associated with lower brain serotonin transporter (5 HTT) binding potential (BP(P), proportional to the number of available transporters) in adults. We examined healthy volunteers and subjects with major depressive disorder, a sample enriched for childhood abuse. METHODS: Regional brain 5-HTT BP(P) was measured using positron emission tomography (PET) with [(11)C]McN 5652 and a metabolite-corrected arterial input function in 43 healthy volunteers and 23 subjects in a major depressive episode, ten of whom reported a history of sexual and/or physical abuse before age 15, and 13 of whom did not. As only two healthy volunteers reported childhood abuse, primary analyses were restricted to the depressed sample, with healthy controls presented as comparators. RESULTS: Depressed subjects reporting childhood abuse had lower 5 HTT BP(P) than nonabused depressed subjects across all brain regions examined (P = 0.017). The groups did not differ in relevant demographic or clinical variables. Genotype frequencies of a functional polymorphism in the 5-HTT gene promoter (5-HTTLPR) did not differ between the groups. CONCLUSIONS: Reported childhood abuse is associated with lower 5-HTT BP(P) in this sample of subjects with major depression, consistent with other reports that childhood adversity can lower serotonergic function permanently. Lower 5-HTT BP(P) may represent a biological pathway through which early life stress predisposes to the development of subsequent psychiatric illness, including major depressive disorder. PMID- 19288579 TI - Serum creatinine ratio: a novel predictor of mortality after percutaneous coronary intervention in patients with normal and abnormal renal function. AB - The occurrence of contrast induced nephropathy (CIN) is associated with increased mortality after percutaneous revascularization procedures. However, the exact correlation between various levels of creatinine elevation relative to the baseline and subsequent mortality in patients with chronic renal insufficiency (CRI) is not well established. In addition, the relationship between elevated postprocedural creatinine and ensuing mortality in patients with normal baseline renal function needs to be investigated. METHODS: All percutaneous coronary intervention (PCI) patients (n = 12,997) were analyzed for any rise in serum creatinine (SCr): CRI group (BSC > or = 1.5 mg/dl) (n = 1,853) and normal baseline renal function (NBR BSC < 1.5 mg/dl) group (n = 11,144). Patients in each group were analyzed for any elevation in SCr postprocedure and subdivided based on the SCr ratio [peak SCr/Baseline creatinine (BSC)] of <1.25, 1.25-1.5, and >1.5. The overall incidence of CIN (defined as an increment of 25% over baseline creatinine) was 5.9%: 11.3% in the CRI group versus 5.1% in normal BSC group (P < 0.01). Recursive partitioning and Cox hazard modeling were used to assess significant variables associated with mortality within 1 year. Only serum creatinine ratio (SCrR) > 1.5 correlated with increased mortality in both CRI group as well as normal BSC group. CONCLUSIONS: SCrR > 1.5 predicts mortality at 1 year after PCI. The association between SCrR > 1.5 and increased mortality at follow-up is observed in patients with CRI as well as normal baseline renal function. SCrR may thus serve as a useful clinical tool for risk stratification and prognostication of patients after PCI. PMID- 19288580 TI - Injury-induced sequential transformation of notochordal nucleus pulposus to chondrogenic and fibrocartilaginous phenotype in the mouse. AB - Intervertebral disc degeneration has been widely studied in different animal models. To test the hypothesis that needle puncture could induce progressive biochemical and molecular changes in murine discs, we established a mouse tail model to investigate the pathogenesis and molecular mechanism of puncture-induced disc degeneration. Caudal discs in mouse tails were punctured using a 31G gauge needle at controlled depth under microscopic guidance. The progress of the disc degeneration was evaluated by radiographic analysis of disc height, histological grading and glycosaminoglycan (GAG) quantification pre-operation and 1, 2, 6 and 12 weeks post-puncture. Gene and protein expression of the extracellular matrix (ECM) was analysed by RT-PCR, in situ hybridization and immunohistochemistry. Histological study and disc height analysis revealed progressive degenerative changes in the punctured discs. Compared with the pre-operation control group, total GAG content decreased 40% (p < 0.05) and aggrecan (Acan), decorin (Dcn) and versican (Vcan; Cspg2) expression was down-regulated at 12 weeks post-puncture. A transient increase of Col2a1-expressing cells and elevation of collagen II protein in the nucleus pulposus (NP) was detected. Fibronectin (Fn1) expression was up-regulated 50% and deposition of collagen I in NP was observed at 12 weeks post-puncture. This study is the first to use an injury-induced model to study disc degeneration in mouse. The disc degeneration involves a transient transformation of NP from notochordal to chondrogenic and eventually into fibrocartilaginous phenotype. The degenerative changes have some similarity to human disc degeneration, suggesting that this model may potentially be used in future to study the molecular mechanism and dissect the pathways of disc degeneration. PMID- 19288581 TI - Association between quality of life and self-stigma, insight, and adverse effects of medication in patients with depressive disorders. AB - BACKGROUND: The aims of this study were to examine whether different domains of quality of life (QOL) are differently affected by depressive disorders by comparing QOL of subjects with and without depressive disorders, and to examine the association of QOL with self-stigma, insight and adverse effects of medication among subjects with depressive disorders. METHOD: The QOL on the four domains of the WHOQOL-BREF Taiwan version were compared between the 229 subjects with depressive disorders and 106 control subjects. Among the depressive subjects, the association between the four QOL domains and subjects' self-stigma, insight, and adverse effects of medication were examined using multiple regression analyses by controlling for the influence of depression, socio demographic and clinical characteristics and family function. RESULTS: Depressive subjects had poorer QOL on the physical, psychological and social relationship domains than the non-depressive control group. The depressive subjects who had more severe self-stigma had poorer QOL on all four domains. The depressive subjects who perceived more severe adverse effects from medication had poorer QOL on the physical, psychological and environmental domains. However, insight was not associated with any domain of QOL in patients with depressive disorders. CONCLUSIONS: The results of this study demonstrate that different domains of QOL are differently affected by depressive disorders, and that clinicians must consider the negative influences of self-stigma and adverse effects from medication on QOL of subjects with depressive disorders. PMID- 19288582 TI - Abnormal neuroendocrine response to clomipramine in hereditary affective psychosis. AB - BACKGROUND: Blunting of prolactin response after serotonergic stimulation during a major depressive episode has been described by several investigators. In this study, the neuroendocrine responses to clomipramine were assessed in remitted patients suffering from hereditary depression. METHODS: Twenty remitted patients from 11 large families with multigenerational, multiple cases of major affective disorder (bipolar disorder n=15, recurrent depression n=5, according DSM-IV) and 12 healthy relatives were investigated. After intravenous application of 12.5 mg of the serotonin re-uptake inhibitor clomipramine, serum prolactin and cortisol levels were analysed. RESULTS: Patients and comparison group did not differ significantly with respect to age, baseline prolactin and cortisol concentrations. A gender effect was found in an exploratory analysis for prolactin but not for cortisol and therefore the data for prolactin were analysed separately. After clomipramine infusion, the increase of cortisol was significantly lower in patients than in the comparison group (P=.046). For prolactin, this effect could be found in the male (P=.012) as well as in the female (P=.007) subsample. CONCLUSIONS: These results suggest that blunted prolactin and cortisol responses to serotonergic stimulation are characteristic for remitted depressive patients with previous episodes of major affective disorders. PMID- 19288583 TI - A population-based family study of minor depression. AB - BACKGROUND: There is a paucity of research on whether minor depression is a familial disorder. METHODS: We conducted a population-based family study of minor depression in which subjects were interviewed using the Diagnostic Interview Schedule (DIS). Minor depression only (MDO) was diagnosed if there was a lifetime history of what the DIS refers to as a depressive "spell" and no lifetime history of either Diagnostic and Statistical Manual of Mental Disorders, Third Edition, major depression or dysthymia. There were 71 probands with a lifetime history of MDO; 577 controls with no lifetime history of MDO, major depression, or dysthymia; and 1,539 first-degree relatives (FDRs). Logistic regression was performed with the presence/absence of MDO in a proband/control as the "exposure" and MDO in an FDR as the "outcome". RESULTS: The odds ratio for the association between MDO in a proband and MDO in an FDR, after adjusting for age and sex of the FDR, was 1.55 (95% confidence interval: 0.93-2.58; P=.093). The study had 80% power to detect an odds ratio as small as 1.97, which is in the range of odds ratios seen for the familial aggregation of major depression. CONCLUSIONS: MDO does not appear to be a familial disorder, raising questions about the validity of "minor depression" as a distinct psychiatric syndrome. PMID- 19288584 TI - Noncompartmental pharmacokinetics analysis of glucose-stimulated insulin response in African-American and Caucasian youths. AB - The objective of this study was to examine the differences in glucose and insulin responses between African-American and Caucasian youths and to determine the associations of between-group differences with sex, body mass index (BMI) and pubertal status using a noncompartmental pharmacokinetic approach. Sixteen African-American and 22 Caucasian healthy adolescents were tested using the frequently sampled intravenous glucose tolerance test. Longitudinal t-tests across each observation revealed that (1) African-American youths have higher insulin concentrations between 4 to 19 min; (2) insulin levels remained similar as subjects were grouped according to sex and pubertal status; (3) for glucose, the only difference was found as it approached steady-state basal level (>100 min) between groups with different BMIs. Linear regression showed that insulin concentrations between 4 to 19 min are associated with BMI in Caucasians. African American youths were found to have higher insulin responses after glucose stimulation and the insulin concentrations were more related to BMI in Caucasians compared with African-Americans. BMI also has a significant effect on the glucose steady state basal level. The acute insulin response to glucose (AIR(g)) extended to 20 min resulted in a more significant racial difference (p<0.0006) compared with the calculation done over 10 min suggested in the past (p<0.001). PMID- 19288585 TI - Comparative proteomic study of two closely related ovarian endometrioid adenocarcinoma cell lines using cIEF fractionation and pathway analysis. AB - The proteomic profiles from two distinct ovarian endometrioid tumor-derived cell lines, (MDAH-2774 and TOV-112D) each with different morphological characteristics and genetic mutations, have been studied. Characterization of the differential global protein expression between these two cell lines has important implications for the understanding of the pathogenesis of ovarian endometrioid carcinoma. In this comparative proteomic study, extensive fractionation of peptides generated from whole-cell trypsin digestion was achieved by coupling cIEF in the first dimensional separation with capillary LC (RP-HPLC) in the second dimensional separation. Online analysis was performed using tandem mass spectra acquired by a linear ion trap mass spectrometer from triplicate runs. A total of 1749 and 1955 proteins with protein probability above 0.95 were identified from MDAH-2774 and TOV-112D after filtering through Peptide Prophet/Protein Prophet software. Differentially expressed proteins were further investigated by ingenuity pathway analysis (IPA) to reveal the association with important biological functions. Canonical pathway analysis using IPA demonstrates that important signaling pathways are highly associated with one of these two cell lines versus the other, such as the PI3K/AKT pathway, which is found to be significantly predominant in MDAH-2774 but not in TOV-112D. Also, protein network analysis using IPA highlights p53 as a central hub relating to other proteins from the connectivity map. These results illustrate the utility of high throughput proteomics methods using large-scale proteome profiling combined with bioinformatics tools to identify differential signaling pathways, thus contributing to the understanding of mechanisms of deregulation in neoplastic cells. PMID- 19288586 TI - A proposed metabolic strategy for monitoring disease progression in Alzheimer's disease. AB - A specific, sensitive and essentially non-invasive assay to diagnose and monitor Alzheimer's disease (AD) would be valuable to both clinicians and medical researchers. The aim of this study was to perform a metabonomic statistical analysis on plasma fingerprints. Objectives were to investigate novel biomarkers indicative of AD, to consider the role of bile acids as AD biomarkers and to consider whether mild cognitive impairment (MCI) is a separate disease from AD. Samples were analysed by ultraperformance liquid chromatography-MS and resulting data sets were interpreted using soft-independent modelling of class analogy statistical analysis methods. PCA models did not show any grouping of subjects by disease state. Partial least-squares discriminant analysis (PLS-DS) models yielded class separation for AD. However, as with earlier studies, model validation revealed a predictive power of Q(2)<0.5 and indicating their unsuitability for predicting disease state. Three bile acids were extracted from the data and quantified, up-regulation was observed for MCI and AD patients. PLS DA did not support MCI being considered as a separate disease from AD with MCI patient metabolic profiles being significantly closer to AD patients than controls. This study suggested that further investigation into the lipid fraction of the metabolome may yield useful biomarkers for AD and metabolomic profiles could be used to predict disease state in a clinical setting. PMID- 19288587 TI - Microfluidic chips for protein differential expression profiling. AB - Biomarker discovery and screening using novel proteomic technologies is an area that is attracting increased attention in the biomedical community. Early detection of abnormal physiological conditions will be highly beneficial for diagnosing various diseases and increasing survivability rates. Clearly, progress in this area will depend on the development of fast, reliable, and highly sensitive and specific sample bioanalysis methods. Microfluidics has emerged as a technology that could become essential in proteomics research as it enables the integration of all sample preparation, separation, and detection steps, with the added benefit of enhanced sample throughput. The combination of these advantages with the sensitivity and capability of MS detection to deliver precise structural information makes microfluidics-MS a very competitive technology for biomarker discovery. The integration of LC microchip devices with MS detection, and specifically their applicability to biomarker screening applications in MCF-7 breast cancer cellular extracts is reported in this manuscript. Loading approximately 0.1-1 microg of crude protein extract tryptic digest on the chip has typically resulted in the reliable identification of approximately 40-100 proteins. The potential of an LC-ESI-MS chip for comparative proteomic analysis of isotopically labeled MCF-7 breast cancer cell extracts is explored for the first time. PMID- 19288588 TI - Candidate gene copy number analysis by PCR and multicapillary electrophoresis. AB - Genetic polymorphisms are often considered as risk factors of complex diseases serving as valuable and easily detectable biomarkers, also stable during the whole lifespan. A novel type of genetic polymorphism has been identified just recently, referred to as gene copy number variation (CNV) or copy number polymorphism. CNV of glycogen synthase kinase 3 beta and its adjacent gene, Nr1i2 (pregnane X receptor isoform), has been reported to associate with bipolar depression. In our study we introduced multicapillary electrophoresis for gene copy number analysis as an affordable alternative to real-time PCR quantification with TaqMan gene probes. Our results show the reliability of the developed method based on conventional PCR followed by separation of products by multicapillary electrophoresis with quantitative evaluation. This method can be readily implemented for the analysis of candidate gene CNVs in high throughput clinical laboratories and also in personalized medicine care of depression-related risk factors. PMID- 19288589 TI - Proteomic analysis of osteogenic differentiation of dental follicle precursor cells. AB - Recently, there has been an increased interest in unravelling the molecular mechanisms and cellular pathways controlling the differentiation and proliferation of human stem cell lines. Proteome analysis has proven to be an effective approach to comprehensive analysis of the regulatory network of differentiation. In the present study we applied 2-DE combined with capillary-LC MS/MS analysis to profile differentially regulated proteins upon differentiation of dental follicle precursor cells (DFPCs). Out of 115 differentially regulated proteins, glutamine synthetase, lysosomal proteinase cathepsin B proteins, plastin 3 T-isoform, beta-actin, superoxide dismutases, and transgelin were found to be highly up-regulated, whereas cofilin-1, pro-alpha 1 collagen, destrin, prolyl 4-hydrolase and dihydrolipoamide dehydrogenase were found to be highly down-regulated. The group of up-regulated proteins is associated with actin bundling and defence against oxidative cellular stress, whereas down-regulated proteins were associated with collagen biosynthesis. Bioinformatic analyses of the entire data set confirmed these findings that represent significant steps towards the understanding of DFPC differentiation. The bioinformatic analyses suggest that proteins associated with cell cycle progression and protein metabolism were down-regulated and proteins involved in catabolism, cell motility and biological quality were up-regulated. These results display the general physiological state of DFPCs before and after osteogenic differentiation. We also identified regulatory proteins, such as the transcription factors TP53 and Sp-1, associated with the differentiation process. Further studies will investigate the impact of identified regulatory proteins for cell proliferation and osteogenic differentiation in DFPCs. PMID- 19288590 TI - Fluorescent isotope-coded affinity tag 2: peptide labeling and affinity capture. AB - Fluorescent isotope-coded affinity tag (FCAT) is a novel reagent to label cysteine-containing peptides. The fluorescein group on the tag enables absolute quantification by fluorescence detection, also supporting affinity capture of the labeled peptides. In this paper we report the synthesis of the heavy isotopic form of the FCAT reagent and its use in labeling tryptic peptides. The heavy form of the reagent exhibited the same reactivity and chromatographic behavior that of the light version. Effective labeling of tryptic peptides from alpha-lactalbumin, fetuin, BSA and phosphorylase b was attained by using both the heavy and light FCAT tags. Selective capture of the FCAT-labeled peptides was easily performed by pipette tips containing either anti-FITC antibody or iminodiacetic-acid-coated beads. The differently labeled peptides were separated by RP HPLC and analyzed by MALDI-TOF MS. PMID- 19288591 TI - [Organ-sparing surgery for testicular tumors]. PMID- 19288592 TI - Robotic-assisted laparoscopic pelvic lymphadenectomy for bladder cancer: a surgical atlas. AB - INTRODUCTION: Recently, robotic approaches to cystectomy have been reported. Lymphadenectomy remains an important diagnostic component of cystectomy. As with resection of the primary tumor, the use of new technologies must not compromise the oncological benefits of lymphadenectomy, We describe our approach to and results of robotic-assisted laparoscopic pelvic lymphadenectomy in cystectomy. TECHNIQUE: We describe the technique of standard and extended pelvic lymph node dissection during robotic-assisted cystectomy. The classic da Vinci or the da Vinci S robotic platform is utilized for lymphadenectomy. RESULTS: Twenty-eight patients underwent a standard dissection with a mean number of lymph nodes removed of 19 (range, 8-33). Extended lymph node dissection has been performed in 22 patients with a mean of 30 lymph nodes removed (range, 12-39). No surgical complications have occurred related to the lymphadenectomy. CONCLUSIONS: Robotic laparoscopic pelvic lymphadenectomy is feasible and safe, and is equivalent in efficacy to open approaches in bladder cancer. PMID- 19288593 TI - Retraction notice to: Fuzzy controller training using particle swarm optimization for nonlinear system control. PMID- 19288594 TI - Biochemistry and molecular biology. [editorial]. PMID- 19288595 TI - Not a myth. PMID- 19288596 TI - Identifying Parkinson's. PMID- 19288597 TI - Half a century of service. PMID- 19288598 TI - Healers helping healers. PMID- 19288599 TI - [The use of laseropuncture in patients with bronchial asthma and concomitant chronic rhinosinusitis]. PMID- 19288600 TI - [Laserotherapy and mesodiencephalic modulation after hemihepatoectomy in the early postoperative period]. PMID- 19288601 TI - [Use of sodium hydrocarbonate mineral water in patients with metabolic syndrome]. PMID- 19288602 TI - [Use of ultrahigh frequency electromagnetic radiation for the treatment of lipoid necrobiosis in children with type 1 diabetes mellitus]. PMID- 19288603 TI - [Principles of management in the development of an integrated information system for a sanatorium-and-spa complex]. PMID- 19288604 TI - [Correction of motor disturbances of central organic origin in children with the use of a novel technology "the theater of recovering and educational motion"]. PMID- 19288605 TI - Normal-like breast cells, but not breast cancer cells, recovered from treatment with N',N''-diethylnorspermine. AB - A number of polyamine analogs are currently used in various clinical trials as cancer treatment and it is important to investigate their effects not only on cancer cells but also on normal cells. Treatment with polyamine analogs depletes cells of polyamines and inhibits cell proliferation, but the analogs cannot take over the normal function of the natural polyamines in the cell. In this study, the normal-like breast epithelial cell line MCF-10A was treated with the polyamine analog N',N"-diethylnorspermine (DENSPM). The cells were then studied using a bromodeoxyuridine- DNA flow cytometry method as well as western blot. The ability of both normal-like and breast cancer cells to recover from DENSPM treatment was also studied. DENSPM treatment of MCF-10A cells resulted in a prolongation of the S and G2 +M phases, followed by a G1/S block. The p53/p21/RB1 pathway was involved in the G1/S block as shown by increased levels of p53 and p21 detected by western blot. Decreased levels of cyclin E1, cyclin A2, and cyclin B1 in DENSPM-treated cells can explain the prolongation of cell cycle phases that occurred before the G1/S block. We also show that MCF-10A cells rapidly recover from DENSPM-induced growth inhibition in contrast to four human breast cancer cell lines. The goal of cancer treatment is to cause minimal and reversible damage to normal cells, while cancer cells should be eliminated. Altogether, the data show that treatment with polyamine analogs spares normal cells, while negatively affecting the cancer cells. PMID- 19288606 TI - Emerging strategies in regulatory T-cell immunotherapies. AB - The ability of tumors to evade immune system surveillance is an important characteristic that allows for their growth and survival. Accumulating evidence suggests that the mechanisms which allow for tumor cell evasion are similar to those that exits to prevent autoimmune diseases. Thus, targeting the immune system may be a major strategy to develop novel anti-tumor therapies. Among these strategies are therapies directed against regulatory T cells, as well as therapies designed to enhance the immune response triggered by CD8+ T cells. In the clinical roundtable, several experts discuss emerging strategies in regulatory T-cell immunotherapies. First, the major concepts behind immune suppression of solid tumors are described, including the current challenges and therapeutic targets under investigation. Then, the strategies under development are examined for their ability to overcome the regulatory T-cell effects in tumors. Finally, the roundtable concludes with a discussion of how these basic immunomodulatory concepts are being translated into clinical application. By understanding this roundtable, the clinician or oncologist will have a strong understanding of the current state-of-the-art strategies under investigation for anti-tumor immunotherapy. PMID- 19288607 TI - [Treatment of labyrinthine diseases--the frontier of experimental studies- Introduction of genes (regeneration of cochlear hair cells and restoration of auditory function)]. PMID- 19288608 TI - Decreasing the increase. PMID- 19288609 TI - This is the greatest generation. PMID- 19288610 TI - The effect of cyclooxygenase-2 inhibition on acute and chronic donor-site pain after spinal-fusion surgery: erratum retraction. PMID- 19288611 TI - Tissue-specific modulation of a set of related cell surface antigens in Drosophila. PMID- 19288613 TI - [Topics from other other medical fields--drug metabolism and drug interactions]. PMID- 19288612 TI - [Treatment of acute pharyngo-tonsillitis using using anti-bacterial agents- current concepts based on meta analysis]. PMID- 19288614 TI - Mandatory reassignment under the ADA: the circuit split and need for a socio political understanding of disability. PMID- 19288615 TI - Health law - genetics - Congress restricts use of genetic information by insurers and employers. - Genetic Information Nondiscrimination Act of 2008, Pub. L. No. 110-233, 122 Stat. 881 (to be codified in scattered sections of 26, 29, and 42 U.S.C.). PMID- 19288616 TI - State constitutional law - same-sex relations - Supreme Court of Michigan holds that public employers may not provide healthcare benefits to same-sex domestic partners of employees. - National Pride at Work, Inc. v. Governor of Michigan, 748 N.W.2d 524 (Mich. 2008). PMID- 19288617 TI - Playing games with girls' health: why it is too soon to mandate the HPV vaccine for pre-teen girls as a prerequisite to school entry. PMID- 19288618 TI - The best laid plans: disappointments of the National Youth Anti-Drug Media Campaign. AB - As part of its war on drugs, the U.S. government spent nearly $1 billion between 1998 and 2004 for the National Youth Anti-Drug Media Campaign. The campaign had three goals: educating children and teenagers (ages 9 to 18) on how to reject illegal drugs, preventing them from starting drug use, and convincing occasional users to stop. Analyzing the effects of this campaign is important not only for future funding decisions but also for more effective targeting of future efforts. This Issue Brief summarizes a Congressionally-mandated evaluation of the campaign's effects on youths' cognitions and behavior around marijuana use. PMID- 19288619 TI - Paying people to lose weight and stop smoking. AB - Unhealthy behaviors, such as smoking, poor diet, and sedentary lifestyles, account for as much as 40% of premature deaths in the U.S. Although behavioral interventions have the potential to improve health, behavior change is difficult, especially over the long term. Many people have difficulty changing health behaviors because it requires trade-offs between immediate consumption and delayed and often intangible health benefits. Incentives can provide people with immediate and tangible feedback that helps make it easier for them to do in the short term what is in their long-term best interest. This Issue Brief explores the use of financial incentives to motivate and sustain smoking cessation and weight loss. PMID- 19288620 TI - [Treatment of labyrinthine diseases--the frontier of pre-clinical studies. Therapeutic strategy to protect the peripheral vestibular organ]. PMID- 19288621 TI - Many Californians with asthma have problems understanding their doctor. PMID- 19288622 TI - Half a million older Californians living alone unable to make ends meet. PMID- 19288623 TI - One in four California adolescent girls have had human papillomavirus vaccination. PMID- 19288624 TI - "The account between us": honor, reciprocity and companionship in male friendship in the later seventeenth century. PMID- 19288625 TI - Teen dietary habits related to those of parents. PMID- 19288626 TI - The case of the British inquisition: money and women in mid-eighteenth-century London debating societies. PMID- 19288627 TI - Genetic specialists trailing research boom. PMID- 19288628 TI - Maintaining health insurance during a recession: likely COBRA eligibility: an updated analysis using the Commonwealth Fund 2007 Biennial Health Insurance Survey. AB - As the U.S. economic downturn continues and job losses mount, more working Americans are likely to lose access to affordable health benefits subsidized by their employers. Analysis of the 2007 Commonwealth Fund Biennial Health Insurance Survey finds that two of three working adults would be eligible to extend job based coverage, under the 1985 Consolidated Omnibus Budget Reconciliation Act (COBRA) if they became unemployed. Under COBRA, however, unemployed workers would have to pay four to six times their current contribution at a time of sharply reduced income. In fact, the latest national figures indicate that, because of high premiums, only 9 percent of unemployed workers have COBRA coverage. Substantial financial assistance of 75 percent to 85 percent of premiums could help laid-off workers maintain coverage. In addition, expansion of Medicaid and the State Children's Health Insurance Program would benefit low-income, laid-off workers and their families who are ineligible for COBRA. PMID- 19288629 TI - [Oral allergy syndrome]. PMID- 19288630 TI - Total hip replacement. PMID- 19288631 TI - Podiatric medicine and surgery (podiatry). PMID- 19288632 TI - [An extensive survey on suicidal incidents at medical facilities in Japan I- suicidal incidents at psychiatric facilities]. PMID- 19288633 TI - [An extensive survey on suicidal incidence at medical facilities in Japan II- status of suicidal incidents at ordinary hospitals and suggestions for suicide prevention]. PMID- 19288634 TI - [Suicidal incidents at psychiatric departments and civil liability]. PMID- 19288636 TI - [Can psychiatric departments of general hospitals survive by reducing their bed sizes? Various approaches chosen and the predicament faced at Toyama Citizens' Hospital]. PMID- 19288635 TI - [Psychiatric departments at general hospitals facing a crisis situation in northern Tohoku area]. PMID- 19288637 TI - [Ideal and reality for newly created psychiatric departments of general hospitals]. PMID- 19288638 TI - [Is it possible to prescribe resuscitation of general hospital psychiatric departments?]. PMID- 19288639 TI - Establishing patient contracts: as easy as ABC. AB - Due to increasing health insurance premiums, many small business owners are unable to provide benefits to their employees. Some are now opting to negotiate directly with physicians for preventive and acute care services. When properly implemented, these agreements have resulted in huge savings for employers as well as decreased insurance headaches for providers. This article describes how to perform an activity-based cost analysis to decide whether this form of healthcare services is right for your practice. PMID- 19288640 TI - Disability insurance and the physician practice: a primer for physicians and office managers. AB - While your office may be familiar with all of the ins and outs of health insurance, disability insurance claims are complex and difficult to navigate, often deliberately so. When the unthinkable occurs and a claim must be filed, physicians are all too frequently stymied by the response of the insurance company to their claim. This article will provide fundamental information for the physician who needs to file a claim as well the practitioner who comes across a long-term disability insurance claim in his or her practice. PMID- 19288641 TI - Negotiating the best deal with a hospital: if it sounds too good to be true it is. AB - In the face of mounting uncertainty in the healthcare marketplace, including the strong likelihood of substantial healthcare reform looming on the horizon, one fact remains certain ... the time for physician organizations to correct negotiation deficiencies is now. Factually identifying and correcting negotiating deficiencies is the first step in achieving parity in negotiating with hospitals. This article exposes sensitive, rarely addressed deficiencies; uncovers strengths; and presents a strategy to achieve parity. PMID- 19288642 TI - Boost revenue and compliance with correct, consistent modifier 25 application. PMID- 19288643 TI - Funding your EMR: how to get the money you need. AB - Many practices today are considering the purchase of an electronic medical record (EMR) system. "When should we get one? Are we ready for this type of technology in our practice? How will the physicians feel about it? How about the staff? How will it change our work flow? And most of all, who is going to pay for it?" are common questions that arise. In fact, funding the EMR is the number one reason most practices delay the decision to move forward with this type of technology in the office. PMID- 19288644 TI - Strength in numbers: consolidation and collaboration strategies for physicians. AB - Developing a position of strength requires greater numbers of providers, often consolidated as legal structures traditionally not typical of small medical practices and other healthcare entities in the United States. This article addresses market stresses that are causing physicians to move from positions of autonomy to arrangements in which responsibility can be shared with other providers. It offers ways to identify and evaluate the appropriate affiliation model for your practice and market area. PMID- 19288645 TI - Using e-health to improve cholesterol management in primary care practice. AB - While e-health tools have been designed for patient/consumer empowerment and for decision support focusing on healthcare providers, they have had only modest success in primary care settings. Creating complementary e-health tools that provide patient activation and decision support for a common clinical problem such as hyperlipidemia using a behaviorally oriented risk communication strategy has not been previously described. This paper presents the details regarding the development of two complementary software programs: HeartAge, an interactive risk communication program focusing on patient activation placed on a laptop computer in the doctor's waiting room, and a PDA-based, clinical decision support program for lipid management utilizing algorithms recommended by the National Cholesterol Education Program. The efficacy of these programs in improving cholesterol management in five primary care practices is described. PMID- 19288646 TI - Trends in the consumer price index. PMID- 19288647 TI - The paper crisis: from hospitals to medical practices. AB - Hospitals, not unlike physician practices, are faced with an increasing burden of managing piles of hard copy documents including insurance forms, requests for information, and advance directives. Healthcare organizations are moving to transform paper-based forms and documents into digitized files in order to save time and money and to have those documents available at a moment's notice. The cost of these document management/imaging systems can be easily justified with the significant savings of resources realized from the implementation of these systems. This article illustrates the enormity of the "paper problem" in healthcare and outlines just a few of the required processes that could be improved with the use of automated document management/imaging systems. PMID- 19288648 TI - Anatomy of a fraud indictment and acquittal. AB - Just because a physician/practice is innocent of any wrongdoing does not mean that it won't be captured in a net of prosecution. Each practice must look at what it needs to do defensively to both avoid the possibility of prosecution and, if an indictment is handed down, proactively defend itself in a cost-effective and positive fashion without having to negotiate a settlement when no fraud has been committed. This article examines the case of one physician's healthcare criminal indictment and his ultimate acquittal; learn from his experience. PMID- 19288649 TI - The mystery patient: what she expects is no mystery. PMID- 19288650 TI - DEA issues proposed rule on e-prescribing. AB - On June 27, 2008, the Drug Enforcement Administration (DEA) released a Notice of Proposed Rulemaking. In an effort to modernize the regulation of controlled substances, the DEA is proposing an amendment to its regulations that would permit practitioners to write and pharmacies to dispense and archive prescriptions of controlled substances electronically. The new regulations aim to promote accuracy and efficiency. This article discusses that proposed rule. PMID- 19288651 TI - 25 tips for working through language and cultural barriers in your medical practice. AB - The language and cultural barriers facing medical patients with limited English language proficiency pose tremendous challenges and risks. Moreover, medical practices today are more likely than ever to employ individuals whose first language is not English or who do not possess native-like knowledge of American culture. Knowing how to work through the language and cultural barriers you are likely to encounter in your medical practice has become increasingly more important. This article is written by a practice management consultant who has graduate-level linguistics training and second-language teaching credentials and experience. It offers 25 practical tips to help you communicate more effectively with individuals who are outside of your native culture and language. These include easy-to-implement tips about English language pronunciation, grammar, and word choice. This article also suggests what you can do personally to bridge the cultural divide with your patients and co-workers. Finally, this article includes a case study of one Virginia practice in which cultural differences interfered with the practice's smooth operation. It explains how the practice eventually worked through and overcame this cultural obstacle. PMID- 19288652 TI - Translating operational research to the medical community: using "guiding measurements" to improve the quality of healthcare delivery. AB - Despite the ubiquity problems with pay-for-performance and other quality improvement initiatives, there is little in the way of objective evidence that these efforts have improved the quality of care. In part, it may be because the measurements selected are used to "grade" instead of guide improvement efforts. We propose using operational research methods that include how to develop "guiding measurements" to improve care. We show that use of this type of guiding measurements can lead to improved patient understanding, throughput, and satisfaction in a pediatric nephrology ambulatory care clinic, and may have wider applications across the continuum of care. PMID- 19288653 TI - Results of a survey of an online physician community regarding use of electronic medical records in office practices. AB - A recent publication suggested that only 24% of U.S. doctors use electronic medical records (EMRs), and that physicians in group practices, urban settings, and multispecialty groups are more likely than others to use EMRs. If the widespread adoption of EMRs is to be accomplished in keeping with the President's Health IT goal of universal use by 2014, the reasons for EMR under-utilization must be ascertained and addressed. In order to assess current patterns of EMR use and reasons for non-use, physician opinions were surveyed directly, using an online social network of physicians in the United States called Sermo. In February 2008, 893 physician members of Sermo responded to the author's posting about EMR use. Their responses were notable for two reasons. First, EMR adoption (not just electronic billing or prescribing) may actually be higher (67%) than previous reports. Second, the most important barriers to EMR use were high cost, loss of autonomy, and workflow disruption (from both installation and ongoing use). These responses are discussed in the context of recent proposals by both private and public entities that EMR systems be given away for free to any physician who will use them, as well as the potential for patients' online personal health records to permit integration of data across multiple EMR systems. PMID- 19288654 TI - Be a real professional! AB - In an era in which healthcare practices are striving to provide better customer service, reduce staff turnover, increase referrals, and build stronger revenues, we need a standard of excellence upon which to base our expectations, model our behaviors, and hold our team members accountable. This article lays a foundation for a model of professionalism that allows your practice to build a team of people who serve as consistent and energized representatives of your unique vision--no matter how big or small your practice. PMID- 19288655 TI - Hospitals face a difficult future. PMID- 19288656 TI - Taking action against someone removing your patient list. PMID- 19288657 TI - My receptionist is getting too old. PMID- 19288658 TI - Cosmetic dentistry is not a recognized specialty. PMID- 19288659 TI - Financial planning for your practice. PMID- 19288660 TI - The difference between a procedure and a technique. PMID- 19288661 TI - St. Clair County's Community Dental Clinic. PMID- 19288662 TI - A clinical trial of the Nomad portable X-ray unit. AB - OBJECTIVES: To compare image quality of intraoral radiographs made with the Nomad portable X-ray unit (Aribex, Inc.) and with a wall-mounted dental X-ray machine in a clinical setting. METHODS AND MATERIALS: Twelve patients requiring a full mouth radiographic series for diagnostic purposes were recruited for the study, in which half of the radiographs (one side selected randomly) were made with the Nomad and half with a wall-mounted dental X-ray machine. Each individual image was evaluated independently by three reviewers for diagnostic utility and quality, using a three-point scale: 2 = perfect radiograph; 1 = acceptable for diagnostic purposes despite a small error; 0 = unacceptable. A total score was derived for each of the 220 images, each of the 12 patients, and the study population as a whole. RESULTS: The combined quality scores of the three reviewers for each set of radiographs ranged from 22-60 for the Nomad and 21-55 for the control units. Median scores were 33 for the Nomad and 33.5 for the control. The maximum possible score was nine films/half mouth x two points for perfect film x three reviewers = 54 (maximum 66 for 11 films). Three of the films, all on the same patient in a wheelchair, showed motion artifact (two with Nomad, one with control). DISCUSSION: Image quality for radiographs taken with the Nomad and a wall-mounted X-ray machine appears to be similar in a variety of clinical situations. Motion artifact is not a significant issue with the Nomad. PMID- 19288663 TI - Ready, aim... PMID- 19288664 TI - Under our wing. PMID- 19288665 TI - MAG gearing up for pivotal legislative session. PMID- 19288666 TI - ASC exemptions from Georgia's new certificate of need law. PMID- 19288667 TI - Physicians' Care Clinic--a bridge to community health care. PMID- 19288668 TI - Savannah's high school preceptorship program building bridges for 10 years. PMID- 19288669 TI - HIPAA requirement preempting civil discovery rules in malpractice and related suits significant. PMID- 19288670 TI - Composition, purpose and regulatory authority of the composite state board of medical examiners. PMID- 19288671 TI - The 2009 OIG Work Plan: strategies for physician practice compliance. PMID- 19288672 TI - Coumadin therapy: loaded for bear without strong guidelines in place. PMID- 19288673 TI - What's your dementiacap? PMID- 19288674 TI - Do rapid 'superbug' tests pay off? Balance the costs and benefits of leading-edge technology. Interview by Alan Joch. AB - As hospitals become increasingly sensitive to the health and financial consequences of health care-associated infections (HAIs), a new generation of molecular-based testing technologies promises to significantly shorten the time required to identify "superbugs" and other bacterial infections. The leading-edge techniques promise to reduce costs by helping hospitals quickly determine which patients to isolate because they carry active methicillin-resistant Staphylococcus aureus (MRSA) infections, for example, or which ones to release from prophylactic isolation because they ultimately tested negative for a dangerous infection. But diagnostic speed comes at a price--the costs to perform molecular tests are significantly higher than conventional methods. This challenges hospitals to balance health care expenses with medical efficacy, says molecular testing veteran Margie Morgan, Ph.D., scientific director at Cedars Sinai Medical Center, Los Angeles. "The rapid methods can be extreme time savers and possibly help a great deal with the isolation of patients. But some of the tests may cost five times what manual methods might be, so there is a price for seeing so much of a reduction in time," she says. PMID- 19288675 TI - At odds no more. Materials, infection control unite to beat infectious diseases. AB - Open communication between infection control and materials managers plays a vital role in patient care. Cost containment is an obvious issue, as is ensuring patient safety. Where collaboration becomes paramount is when the two departments work together toward standardization while giving infection control the flexibility to buy off-contract when necessary. Both must have an understanding of what products are available and which of those products are going to fit the needs of clinicians and patients. PMID- 19288676 TI - Understanding the effects of sterile processing chemicals. Protection, education are paramount in ensuring worker safety. AB - Central service professionals play a vital role in ensuring patient safety, and in doing so, are exposed to harsh chemicals and sterilants. Knowing the effects of such chemicals is important so that the proper measures can be taken to prevent personal harm or injury as much as possible. Through the use of personal protective equipment, worker training and firmly adhering to OSHA practices, employees can work safely in the central service environment; especially when used in conjuction with engineering controls and gas monitoring. PMID- 19288677 TI - Affecting culture change. Leadership involvement imperative to ensure paradigm shift. AB - As the supply chain evolves, health care organizations must evolve as well. New demands are being made on materials managers, and other departments also are feeling the economic pressure. But change is not possible without the support of leaders, especially those in executive positions. By making a convincing case to those who embrace change, a culture shift can begin to take place. This will unify departments, synchronize organizational goals and create a more efficient decision-making process involving clinicians. PMID- 19288678 TI - The gold standard in sterilizer monitoring. AB - Sterilizer monitoring with biological indicators (BIs) is considered the gold standard since BIs actually test a sterilizer's ability to kill specific strains of highly resistant organisms. Commercially available BIs consist of spores in or on a carrier, accompanied by culture medium with a color change pH indictor dye. Here, SPSmedical Supply Corp. educators address questions about these important quality assurance devices. PMID- 19288679 TI - OR software syncs efficiency, savings. PMID- 19288680 TI - Making sense and success of product conversions and implementations. PMID- 19288681 TI - Acronyms: what's in a name? PMID- 19288682 TI - Through caverns measureless to man. PMID- 19288683 TI - Overweight and obesity in Rhode Island: developing programs to combat the obesity epidemic. PMID- 19288684 TI - Treatment of pediatric obesity. PMID- 19288685 TI - Adult behavioral weight loss treatment. PMID- 19288686 TI - Maintenance of long-term weight loss. PMID- 19288687 TI - Bariatric surgery for severe obesity: the role of patient behavior. PMID- 19288688 TI - Cancer screening over 65. PMID- 19288689 TI - Self-report of sugar-sweetened beverage and fast food consumption by annual household income. PMID- 19288691 TI - Onward and upward with prefixes. PMID- 19288690 TI - Prostate cancer screening practice and knowledge in Rhode Island. PMID- 19288692 TI - Lip augmentation and contour correction with a ribose cross-linked collagen dermal filler. AB - BACKGROUND: Dermicol-P35 is a collagen gel for soft-tissue cosmetic correction. This study evaluated the affects of Dermicol-P35 in patients who underwent lip enhancement. METHODS: This open, multicenter, retrospective data collection study included patients who received an injection of Dermicol-P35 30G to the lip. All patients who had undergone Dermicol-P35 30G lip injections at 3 cosmetic clinics in Europe were identified and evaluated for inclusion in the study. Assessments were conducted by a clinic investigator immediately after the initial injections and after any touch-up injections as well as 4 months to 10 months after the initial injection. Efficacy measures included subjective impression of improvement by the clinic investigator who performed the injections, and subjective satisfaction of both the investigator and patient with treatment. Safety measures included any adverse events, and the occurrence of swelling, bruising, lumpiness, and pain. RESULTS: Fifty-seven patients were identified to have undergone lip injections, and 51 of these patients met criteria for inclusion in the study. Four months to 10 months postinjection, most patients (98%) were believed to have an improvement in lip enhancement, and the clinic investigators rated themselves as "very satisfied" or "satisfied" with 90% of the treatment results. The majority of patients (94%) were either "very satisfied" or "satisfied" with the Dermicol-P35 30G enhancement. The majority of treatments elicited no reports of swelling, bruising, or lumpiness. CONCLUSION: These study results support Dermicol-P35 30G treatment as a safe and effective treatment for lip enhancement. PMID- 19288693 TI - [Opinions of health professionals on the impact of the Preventive Activities and Health Promotion Program (PAPPS)]. AB - OBJECTIVE: To find out the opinion of Primary Health Care (PHC) professionals on the impact of the Preventive Activities and Health Promotion Program (PAPPS). DESIGN: Descriptive qualitative-quantitative study. In a first phase the SWOT technique was used. In a second phase a 47 item questionnaire was designed using an ordinal scale. PARTICIPANTS: Professionals of PAPPS health centre, PAPPS managers and PHC management, using the Autonomous Community, and the time of ascribing or knowledge of PAPPS as segmentation criteria. In the qualitative study 62 participants were selected by theoretical sampling. In the quantitative study, 198 professionals took part in the survey. PRINCIPAL MEASUREMENTS: The assessment of the results has taken into account the responses to the questions formulated and by performing a crossed analysis between strengths/threats and weaknesses/opportunities. A descriptive statistical analysis of the questions in the questionnaire. RESULTS: There is agreement in that PAPPS has greatly influenced the development of PHC, contributing to improving the quality of care, but it is also fundamental to try revitalise the programme, as the limited involvement by the professionals in the postulated preventive recommendations is its main weakness, due to lack of motivation and professional burn-out.. CONCLUSIONS: According to the participants PAPPS has contributed significantly to the development of PHC in our country and has had an influence on professional practice by instilling a prevention culture that hardly existed before. PMID- 19288694 TI - [Preventive cardiovascular recommendations]. PMID- 19288695 TI - [Recommendations on life style]. PMID- 19288696 TI - [Cancer prevention strategies]. PMID- 19288697 TI - [Prevention of infectious diseases]. PMID- 19288698 TI - [Mental health prevention programme in primary care]. PMID- 19288699 TI - [Preventive activities in the elderly]. PMID- 19288700 TI - [Preventive activities in women]. PMID- 19288701 TI - [Child and adolescent programme]. PMID- 19288702 TI - [Comparison of light characteristics in different size gaps in eastern Liaoning montane secondary forests]. AB - With three different size gaps (G1, 670 m2; G2, 290 m2; and G3, 90 m2) in eastern Liaoning montane secondary forests of China as test objects, and through a continuous measurement of light intensity, the spatiotemporal distribution of photosynthetic photon flux density (PPFD) in the gaps was compared. The results showed that the diurnal variation of PPFD in the gaps was greater at northern than at southern position. The larger the forest gap, the broader the higher PPFD value area, and the more obvious the heterogeneity. As for the monthly variation of PPFD, the maximum PPFD at each position of the gaps appeared at the beginning of growth season (i.e., in April and May), while the minimum PPFD occurred in different months. The maximum PPFD at the eastern and western positions of each gap was basically appeared in the same months, and the PPFD was significantly higher in spring than in summer and autumn (P < 0.05). The mean monthly PPFD at the centers of G1, G2, and G3 was 66.59%, 49.05%, and 30.37% of full sunlight, respectively, and in growth season, the PPFD at gap center was 37.8, 27.9 and 10.3 times higher than that at understory. It was suggested that owing to the different size and shape of forest gap and the effects of landform, the light intensity and its distribution in forest gap were different, being the key factors leading to the heterogeneity of regeneration pattern and the variation of species composition in forest gap. PMID- 19288703 TI - [Microclimatic characteristics of different size gaps in Pinus koraiensis dominated broadleaved mixed forests]. AB - By the help of HOBO automatic weather stations, the microclimatic characteristics of different size forest gaps in Pinus koraiensis-dominated broadleaved mixed forests in Xiaoxing' an Mountains were studied during growth season, with closed forest and bare ground as the control. The results showed that the diurnal variation of illumination, ground temperature, and air temperature of the gaps and the controls, as well as the change trend of the microclimatic factors during growth season, all presented a single peak curve. The diurnal variation pattern of relative humidity of the gaps and the control was high-low-high, being higher in early morning and evening and lower at noon. The relative humidity during growth season also presented a single peak curve, and was the highest for closed forest, followed by small, medium, and large forest gaps, and bare ground. The amount and the times of precipitation tended to decrease with increasing forest canopy, and the precipitation amount in large gaps was about 1.4 times as much as that in small gaps. During growth season, the maximum wind speed above bare ground, large, medium, and small gaps, and closed forest was 3.34, 2.97, 2.87, 2.41 and 1.84 m x s(-1), respectively. PMID- 19288704 TI - [Carbon storage maturity age of Larix olgenisis and L. kaempferi]. AB - The successive years and average increments of the volumes of Larix olgensis and L. kaempferi were computed by Trunk Analysis, and the carbon content and carbon density of L. olgensis and L. kaempferi at their different age stages were measured with Germany Vario EL III element analyzer, aimed to approach the carbon storage maturity age of the two tree species. The results indicated that L. olgensis and L. kaempferi had a volume maturity age of 48.3 and 49.3 years, respectively, and the change trend of their carbon density was basically identical. The maximum carbon density was presented in the 30th year for L. olgensis and the 35th year for L. kaempfer. The successive years carbon storage of L. kaempferi was larger than that of L. olgensis, but the carbon storage maturity age was in adverse. The average carbon storage of L. kaempferi was also larger than that of L. olgensis. Through simulating the curve equations of successive years and average carbon storages, the carbon storage maturity age of L. olgensis and L. kaempferi were computed as 48.7 years and 47.7 years, respectively. PMID- 19288705 TI - [Characteristics of seed germination of rare plant species Reaumuria trigyna in west Ordos]. AB - Reaumuria trigyna is a relic species in the desert shrubbery vegetation in arid regions of northwestern China, and plays an important role in the maintenance of the stability of desert vegetation. In this paper, the seed traits and germination strategy of R. trigyna under different environmental conditions, e.g., light, temperature, soil moisture, and sand bury, were investigated. The results showed that R. trigyna seed had high vigor and high germination rate, and endured reserve. The seed could germinate either in light or in darkness, and the optimal temperature for germination was 20 degrees C - 25 degrees C or 15 degrees C/25 degrees C, with the germination rate being 93%. The seed could start to germinate when soil moisture content was 2%, and the germination rate was the highest (89%) when the moisture content was 12%. The optimal sand burial depth of R. trigyna seed was 1 cm, and no seed would germinate when the sand burial depth was >5 cm. Sand burial depth had significant effects on the seedling's emergence percentage and growth height, but lesser effects on seedling' s mass. Soil moisture and sand burial depth were the main environmental factors limiting the seed germination and seedling emergence of R. trigyna. The high seed germination rate of R. trigyna enhanced the survival risk of its seedlings, which was unfavorable to its handling with the extreme changes of desert environment. Such a character of R. trigyna seed was one of the factors causing the species endangered. PMID- 19288706 TI - [Population life process and spectral analysis of rare and endangered plant Alsophila spinulosa]. AB - According to population life table and survival analysis theory, the time specific life table and the survival function curve of Alsophila spinulosa population were established and drawn based on height class, and the life process of the population was analyzed. The results revealed that the population structure of A. spinulosa was of the type ascending at early stage and stable at late stage. The peak of expected life appeared at the 4th age class, indicating the relative good survival quality at this phase, and the survival curve belonged to the typical form of Deevey-III. There existed three death peak phases in the population life process, i.e., the growth stages from 2nd to 3rd age class, from 10th to 11th age class, and from 12th to 13th age class. With the increase of age class, the population survivability dropped down monotonously, and ow nosy, a the accumulative mortality rose up correspondently. The variation extent was high in early phase and low in anaphase. The death density function curve displayed the characteristics of descent in early phase and stabilization in anaphase, and the concave point occurred at 4th age class. Spectral analysis showed that the quantitative fluctuation of A. spinulosa population was multi-harmonic superposition with large cycles containing small cycles. The influences of these cycles dropped with the diminishment of cycle length, and the effect of fundamental wave was most prominent. PMID- 19288707 TI - [Biology and phenology of Xanthoceras sorbifolia in Wudan area]. AB - Based on the continuous observation on located trees and branches, the phenology and sexual reproductive process of Xanthoceras sorbifolia Bunge in an annual growth cycle in Wudan area were investigated, and the relationships between fruit growth process of X. sorbifolia and effective accumulative temperature as well as the quantitative dynamics of flowers and fruits were studied. The results showed that the phenology of X. sorbifolia had a significant linear correlation with effective accumulative temperature, and there was a time sequence in the male flower silk elongating and antheral unfolding. The fruits of X. sorbifolia experienced three times of dropping during their growth process. The pollination rate was 26.4%, flower dropping rate was 73.6%, and fruit dropping rate was 92.9%, suggesting that the dropping of flowers and fruits was very serious. The fruits had a quicker vertical growth from early June to mid June and a quicker horizontal growth in the last ten-day of June, with the highest ratio of vertical to horizontal length being 2.17. The individuals of X. sorbifolia had definite differences in their phenology, which was disadvantageous to the cross pollination among the individuals, and the enhancement of pollination rate. PMID- 19288708 TI - [Relationships of soil moisture content with precipitation and evaporation in rehabilitated forests in degraded limestone red-soil region of Jiangxi Province]. AB - By using time series analysis, the relationships of 0-40 cm soil moisture content with precipitation and evaporation in four main rehabilitated forests in degraded limestone red-soil region of Xiushui County, Jiangxi Province were studied. The results showed that in the four rehabilitated forests, the previous month's soil moisture content had stronger effects on the current month's soil moisture content in 0-40 cm and 20-40 cm layers, but had lesser effects on that in 0-10 cm layer. The soil moisture content in 20-40 cm layer was mainly affected by current month's precipitation, while that in 0-10 cm layer was mainly affected by current month's evaporation. The correlation coefficient between current month's precipitation and soil moisture content was the largest in pure Liquidambar formosana plantation, and the auto-interrelation coefficient of soil moisture content was larger in mixed forests than in pure L. formosana plantation. PMID- 19288709 TI - [Responses of Agriophyllum squarrosum phenotypic plasticity to the changes of soil nutrient and moisture contents and population density]. AB - This paper studied the phenotypic plasticity of Agriophyllum squarrosum under effects of soil nutrient and moisture contents and population density. The results showed that with the increase of soil nutrient content, the root/shoot ratio of A. squarrosum was decreased from 0.135 to 0.073. However, soil moisture content and population density had less effect on the root/shoot ratio. The plasticity of reproductive allocation of A. squarrosum as responding to the changes of soil nutrient and moisture contents was a "real plasticity", and the allocation was negatively correlated with soil nutrient content but positively correlated with soil moisture content. When soil nutrient content was high or moisture content was low, the reproductive allocation of A. squarrosum changed larger with plant size. Population density had no effects on the reproductive allocation, while plant size conditioned the allocation. Among the three test affecting factors, soil nutrient content had the greatest effects on the morphological characters and biomass of A. squarrosum. PMID- 19288710 TI - [Responses of marsh wetland plant Calamagrostis angustifolia to exogenous nitrogen input]. AB - Taking the typical marsh wetland plant Calamagrostis angustifolia in Sanjiang Plain of Northeast China as test material, a field control experiment was conducted to study its leaf morphological characters, chlorophyll content, and aboveground biomass under effects of different levels of exogenous nitrogen input in 2004-2007. Four nitrogen (N) fertilization treatments, i.e., 0 (CK), 6 g x m( 2) x a(-1) (N6), 12 g x m(-2) x a(-1) (N12) and 24 g x m(-2) x a(-1) (N24), were installed. The results showed that there were no significant differences in the leaf length and width among different N fertilization treatments, but the specific leaf area was the smallest (149.54 +/- 18.27 cm2 x g(-1)) under N12, which indicated that the leaves of C. angustifolia were thicker under this N treatment. The leaf chlorophyll content showed a unimodal distribution over the growth season, which increased with the amount of N input before the peak. The peak occurred earlier under N12 and N24 than under N6 and CK. After the peak, the chlorophyll content declined sharply under N24 but slowly under N6, indicating that moderate N input could delay leaf senescence. Continuous high N (N24) input might advance plant growth, while leaf senescence would occur earlier. By the end of growth season both in 2005 and in 2007, the aboveground biomass of C. angustifolia increased with increasing N input, but declined by 53.72% after 4 years continuous high N (N24) input (in 2007), compared with that after 2 years treatment (in 2005). PMID- 19288711 TI - [CO2 exchanges between mangrove- and shoal wetland ecosystems and atmosphere in Guangzhou]. AB - Based on the investigation of biomass and the measurement of CO2 and CH4 fluxes, the CO2 exchanges between mangrove- and shoal wetland ecosystems and atmosphere in Guangzhou were studied, and the CO2 absorption capability of the wetlands vegetation net productivity as well as the carbon sink function of the wetlands under different waterlogged conditions (perennial, intermittent, and no water logging) was analyzed. As for mangrove wetland ecosystem, its vegetation net productivity absorbed 33.74 t x hm(-2) x a(-1) of CO2, and soil emitted 12.26 t x hm(-2) x a(-1) of CO2 (including the greenhouse effect amount of CH4 converted into that of CO2,) illustrating that mangrove wetland had a 21.48 t x hm(-2) x a( 1) net absorption of CO2, being a strong carbon sink. For shoal wetland ecosystem, its vegetation net productivity absorbed 8.54 t x hm(-2) x a(-1) of CO2, and soil emitted 5.88 t x hm(-2) x a(-1) of CO2 and 0.19 t x hm(-2) x a(-1) of CH4. If converting into carbon, the wetland absorbed 2.33 t C x hm(-2) x a( 1), and soil emitted 1.74 t C x hm(-2) x a(-1) (including the carbon in CH4), illustrating that shoal wetland fixed 0.59 t C x hm(-2) x a(-1), being a weak carbon sink. If the greenhouse effect amount of CH4 was converted into that of CO2, the soil emitted 9.78 t x hm(-2) x a(-1) of CO2, which was 1.24 t x hm(-2) x a(-1) more than the absorption. As a result, shoal wetland was a weak carbon source. Between the two test greenhouse gases, CH4 was the main one emitted under perennial water-logging, while CO2 was that under no water-logging. Moreover, the wetland under perennial water-logging had the strongest carbon sink function, while that under no water-logging was in adverse. PMID- 19288712 TI - [Contribution of soil seed bank to the regeneration of damaged vegetation on floodplain]. AB - A field germination experiment of soil seed bank was carried out on two typical floodplains in the lower reaches of Tarim River, and a comparison was made between the soil seed banks and corresponding seedling banks on the two floodplains, aimed to assess the contribution of soil seed bank to the regeneration of damaged vegetation. The results showed that there were 12 plant species in the soil seed banks, and the life forms were mainly perennial herbs and shrubs. The soil seed banks had a density of 282.5 seeds m(-2) and 173.2 seeds x m(-2), and the seeds in top soil (0-2 cm) accounted for 76.9% and 71.0% of the total, respectively. The soil seed banks had significant effects on the seedlings species composition and density, and 84.7% and 99.4% of the seedlings on the two floodplains were emerged from corresponding soil seed banks. The similarity coefficient between soil seed bank and seedling bank of the two floodplains was 0.72 and 0.63, respectively, and there existed significant positive correlation between seedling density and soil seed bank density, illustrating that soil seed bank made important contribution to the natural regeneration of vegetation. PMID- 19288713 TI - [Roles of nitrogen fertilization in regulating the physiological bases of fiber specific strength formation in cotton bolls bloomed at different dates]. AB - To study the roles of nitrogen fertilization in regulating the physiological bases of fiber specific strength formation in cotton bolls bloomed at different dates (July 15, July 25, and September 10; expressed as PSB, SB, and AB, respectively), field experiments were conducted in two sites with different ecological conditions, i.e., Nanjing (middle lower reach of Yangtze River Valley) and Xuzhou (Yellow River Valley). Three treatments (0, 240, and 480 kg N x hm( 2)) were installed. The results showed that comparing with applying optimum nitrogen (240 kg N x hm(-2)), no N application decreased the N content and increased the C/N in the leaves subtending cotton bolls significantly, and the effects increased with postponing blooming date, which, to a great extent, led to the decreases of the capabilities of the leaves subtending cotton bolls bloomed at SB and AB in the accumulation and transportation of photosynthetic products, the relative growth rate of fiber in cotton bolls, and the activities of key enzymes (sucrose synthetase and P-1,3-glucanase) during the middle-later period of fiber development, and also, the decline of cellulose speedily accumulating duration in cotton fiber and of the fiber specific strength in the bolls bloomed at SB and AB. On the contrary, applying high nitrogen (480 kg N x hm-2) increased the N content but decreased the C/N in the leaves subtending cotton bolls significantly, and the effects decreased with postponing blooming date, which decreased the distribution proportion of photosynthetic products to fiber, the key enzyme activities during the early-middle period of fiber development, and the momentary rate of cellulose accumulation in the fiber of the bolls blooming at PSB and SB, resulting in a significant decrease of the fiber specific strength. It was suggested that appropriate N application could regulate the 'source-sink' relation of cotton, and benefit the high fiber specific strength formation in cotton bolls bloomed at different dates. PMID- 19288714 TI - [Effects of post-anthesis irrigation frequency on the grain quality of strong gluten winter wheat cultivars]. AB - In order to investigate the effects of post-anthesis irrigation frequency on the grain quality of strong gluten winter wheat, two cultivars Jimai 20 and Gaocheng 8901 were subjected to a series of irrigation frequencies under rainfall proof conditions, with their grain yield and grain quality (farinograph parameters and loaf volume) and protein composition evaluated. The results indicated that with increasing irrigation frequency, the grain yield of the two cultivars, their wheat flour dough development time, dough stability time, and loaf volume were noted to be increased first but decreased then. The grain yield and quality of Gaocheng 8901 were the highest when irrigated once after anthesis, while those of Jimai 20 were the best when irrigated twice after anthesis, respectively. The contents of monomeric protein, soluble glutenin, insoluble glutenin, total glutenin, flour protein, and wet gluten in the grains displayed the similar trends. Stepwise regression analysis showed that under the test post-anthesis irrigation frequencies, the key factor affecting dough stability time was insoluble glutenin content, and loaf volume was significantly correlated with total glutenin content. It was suggested that to maintain the quality stability of high grade strong gluten winter wheat, irrigation management should take the improvement of grain protein composition, and glutenin in particular, as the target. PMID- 19288715 TI - [Effects of salicylic acid on D1 protein phosphorylation and PS II performance in wheat leaf chloroplasts under high temperature and high light stress]. AB - To study the effects of salicylic acid (SA) on the D1 protein phosphorylation and PS II performance in wheat (Tritivum aestivum L.) leaf chloroplasts under high temperature and high light, the wheat leaves at grain-filling stage were sprayed with SA solution (0.5 mmol x L(-1)) or water (as control), and then subjected to 35 degrees C and 1600 micromol x m(-2) x s(-1) for various hours. The changes in electron transport rate (ETR), net photosynthetic rate, chlorophyll fluorescence parameters, and relative amount of phosphorylated and nonphosphorylated D1 protein in thylakoid membrane were compared. The results showed that foliar spraying SA effectively inhibited the degradation of D1 protein under high temperature and high light stress, and maintained the D1 protein phosphorylation, ETR of whole chain and PS II, Fv/Fm (the maximal photochemical efficiency of PS II), phi(PS II) (the actual photochemical efficiency of PS II), q(p)(the photochemical quenching coefficient), and Pn (net photosynthetic rate) at a higher level, indicating that exogenous SA could mitigate the damage effect of high temperature and high light on wheat leaf photosynthetic apparatus and benefit PS II performance via regulating the turnover of D1 protein in chloroplasts. PMID- 19288716 TI - [Effects of non-sufficient irrigation with saline water on soil water-salt distribution and spring corn yield]. AB - A field experiment was conducted to study the effects of sufficient and non sufficient irrigation with saline water on the soil water-salt distribution and spring corn yield in the middle reach of Shiyanghe River Basin. The results showed that under both sufficient and non-sufficient irrigation, the peak value of soil water content all appeared during irrigation period, and the variation range of the water content was higher under sufficient than under non-sufficient irrigation. Soil salinity was positively correlated with the salinity of irrigation water. At the same salinity of irrigation water, the soil salinity under non-sufficient irrigation was lower than that under sufficient irrigation. Under non-sufficient irrigation, the soil layer with salt accumulation was moved up, but the water and salt contents in 80-100 cm soil layer were less affected by the amount and salinity of irrigation water. Comparing with that under fresh water irrigation, the spring corn yield under saline water irrigation was decreased by 15%-22%. Under non-sufficient irrigation with 9 g x L(-1), 6 g x L( 1), and 3 g x L(-1) of saline water, the average salt content in 1 m soil layer after harvest was decreased by 8.1%, 12.4%, and 18.4%, and the corn yield was only decreased by 3.4%, 6.8%, and 3.0%, respectively, compared with those under sufficient irrigation. PMID- 19288717 TI - [Effects of temperature regime on low-light tolerance of Cucumis sativus seedling leaves in their photosynthesis]. AB - In a phytotron, the effects of three temperature regimes (day/night 25 degrees C/18 degrees C, optimal temperature; 15 degrees C/9 degrees C, suboptimal temperature; and 9 degrees C/7 degrees C, low temperature) on the low-light (75 85 micromol x m(-2) x s(-1)) tolerance of two Cucumis sativus cultivars (shade susceptible Jinyan 2 and shade-tolerant Deltastar) seedling leaves in their photosynthesis were studied. The results showed that under low light, the SPAD, net photosynthesis rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), water use efficiency (WUE), actual photochemical efficiency of PS II in the light (phi(PS II)), and photochemical quenching of chlorophyll fluorescence (q(p)) of cucumber leaves decreased, with the decrement getting more with decreasing temperature, while the superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities were in adverse. During the recovery process after low-light stress relieved, the parameters of gas exchange and chlorophyll fluorescence of the leaves recovered gradually, and the recovery of some gas exchange parameters lagged to that of chlorophyll fluorescence parameters. Under low light, the lower the temperature, the more damage the photosynthesis apparatus suffered, and the lesser tolerance to low light the cucumber leaves had in their photosynthesis. During the low temperature and low light treatment period, the decrease of Pn, phi(PS II), and q(p) was more obvious for Jinyan 2 than for Deltastar; and during the relief period, the recovery of these parameters was slower for Jinyan 2 than for Deltastar. It was indicated that Jinyan 2 had weaker tolerance to low temperature and/or low light in its photosynthesis than Deltastar. PMID- 19288718 TI - [Analysis on quantitative trait loci associated with leaf chlorophyll content and their interactions with environment at late growth stage of rice]. AB - By using a population of 123 F12 lines (recombinant inbred lines, RILs) derived from a cross between India variety Dular and Japanica variety Lemont, an analysis of quantitative trait loci (QTL) was conducted for the flag leaf chlorophyll content of rice. The chlorophyll content was determined by SPAD-502 in 2005 and 2006, respectively, and software QTLMapper 1.6 was applied to analyze the QTL, including the additive and epistatic effects and the QTL interactions with environment for chlorophyll content. A total of ten QTL showing additive effects on chlorophyll content (Chl) were detected, which accounted for 73.51% of the phenotypic variation. The percentage of phenotypic variation explained by single QTL was 2.08%-20.14%, and the interactions of 6 QTLs with environment (AE) were significant. Epistasis analysis indicated that there existed 13 significant additive x additive interactions on chlorophyll content, 6 pairs of which were significant in additive x additive interactions with environment (AAE). PMID- 19288719 TI - [Effects of graft and nitrogen supply level on water consumption and water use efficiency of solar greenhouse cucumber under traditional irrigation]. AB - An experiment was conducted in solar greenhouse to study the effects of graft and different nitrogen (N) supply levels (0, 110, and 331 kg k x hm(-2)) on the water consumption and water use efficiency (WUE) of cucumber (Cucumis sativus L.) under traditional irrigation. The results showed that when topdressing 331 kg N x hm( 2), the water consumption of grafted cucumber was the highest, being 3350 m3 x hm(-2) and 2181 m3 x hm(-2) in winter-spring and autumn-winter cropping, so did the WUE, being 27.2 kg x m(-3) and 36.9 kg x m(-3), respectively. Comparing with those of self-rooted cucumber, the water consumption, economic yield, and WUE of grafted cucumber under the same N topdressing levels were increased by 3%-6%, 28%, and 20.9%, respectively, and the water consumption and WUE of grafted cucumber were increased with increasing N supply level. The water consumption of self-rooted cucumber also increased with increasing N supply level, but its WUE was the highest when the N supply was 110 kg graft could markedly promote the water consumption and WUE of cucumber, and the effect was more markedly with increasing N supply level. To increase N supply could also markedly promote the water consumption of self-rooted cucumber, but the WUE would be lower when excessive N was supplied. PMID- 19288720 TI - [Strong stem effect and physiological characteristics of rice plant under rice duck farming]. AB - A field plot comparison experiment was conducted to study the strong stem effect and physiological characteristics of rice plant under rice-duck farming. The results showed that under rice-duck farming, the morphology of rice plant changed obviously, and the carbohydrate content and C/N ratio of the plant as well as the dry matter output from rice stem increased significantly, due to the activities of duck in paddy field. In the meantime, the length of basal internodes decreased by 2. 88%, while the stem diameter, stem mechanical strength, and stem lodging resistant index increased by 64.90%, 11.78%, and 10.95%, respectively. Rice-duck farming increased the root mass and root vitality in deeper soil layers, and decreased the proportion of black roots by 16.63%. It was indicated that rice duck farming benefited the formation of strong rice stem and increased the stress resistance of rice plant, which would be favorable to the stable and high yielding of rice. PMID- 19288721 TI - [Effects of rice-duck farming on paddy field's methane emission]. AB - By using closed-chamber technique, the characteristics of CH4 emission from an integrated rice-duck ecosystem were studied in 2006 and 2007. The results showed that comparing with conventional flooded rice ecosystem (CK), integrated rice duck system (RD) was distinguished in its high dissolved oxygen (DO) in field surface water and significantly lower CH4 emission, due to the activities of duck. In 2006, the average CH4 emission flux and the total CH4 emission during the whole rice growth period of RD were 6.84 +/- 1.49 mg +/- m(-2) x h(-1) and 19.34 +/- 1.15 g x m(-2), being 32.7% and 26.3% lower than those of CK (10.17 +/- 1.25 mg x m(-2) x h(-1) and 26.25 +/- 2.17 g respectively; while in 2007, the values of the two test items of RD were 7.68 +/- 0.74 mg x m(-2) x h(-1) and 18.41 +/- 1.05 g x m(-2), being 19.0% and 19.3% lower than those of CK (9.53 +/- 0.40 mg x m(-2) x h(-1) and 22.81 +/- 0.75 g +/- m(-2)), respectively. The emission flux of CH4 had two peaks, being appeared at tillering and heading stages, respectively. The seasonal fluctuations of CH4 emission flux had significant positive correlations with soil temperature and soil dissolved organic carbon content (DOC), but no obvious correlation with soil total organic carbon. PMID- 19288722 TI - [Analysis on urban spatial expansion process in Shenyang City in 1979-2006]. AB - Based on the 1979, 1992, 2001 and 2006 satellite images, the spatial expansion information of Shenyang City was extracted by human-computer interactive method, and the temporal and spatial expansion characteristics of Shenyang City as well as their driving forces were analyzed from the aspects of growth intensity, center coordinates, compact index, fractal index, and elasticity coefficient, with the help of the statistic and analytic functions of GIS for the spatial data. The results showed that in 1979-2006, both the urban build-up area and its expansion rate in Shenyang were increased continuously, and reached the maximum in 2001-2006. The urban expansion showed obvious spatial heterogeneity, with the gravity shifted to the southwest. In the meantime, the compact index was decreasing while the fractal index was increasing, implying that the urban spatial configuration in Shenyang became more and more complex. Natural environmental factors, economic development, population growth, traffic infrastructure construction, and government policy and planning were the main driving forces of the urban expansion. PMID- 19288723 TI - [Differences in urbanization process of catchments in dongjiang watershed and their effects on landscape pattern]. AB - Based on 1991, 1998, and 2006 TM images, the areas of different land use types and the landscape indices of three catchments (catchment a, b, and c, which represented upper, middle, and lower reaches, respectively) in Dongjiang watershed were analyzed, aimed to study the differences in urbanization process along Dongjiang River, and their effects on landscape pattern. The results showed that the degree and speed of urbanization increased from the upper to the lower reach of Dongjiang River. Urbanization had significantly effects on water and vegetation. Urban land area was positively correlated with water body area, and negatively correlated with forest land area. However, to some extent, urbanization stepped into a relatively high degree might benefit forest recovery. The landscape pattern of catchments a and b kept complicating from 1991 to 2006, while that of catchment c was getting complex from 1991 to 1998 and then becoming simple from 1998 to 2006, indicating that with the development of urbanization, landscape pattern presented a "simple-complex-simple" tendency. Understanding the change patterns of the landscape pattern along Dongjiang River would benefit the management and sustainable development of the watershed as a whole. PMID- 19288724 TI - [Dynamic changes of groundwater level and vegetation in water table fluctuant belt in lower reaches of Heihe River: coupling simulation]. AB - Based on the 2006 investigation data in lower reaches of Heihe River, and by using logarithmic normal distribution model, the models about the vegetation cover of Populus euphratica and Tamarix ramossima and the groundwater level in study area were built, and the potential plant of the study area was simulated. The results showed that in the lower reaches of Heihe River, the optimal groundwater level and mean groundwater level for P. euphratica were 2.6 m and 3.6 m, and those for T. ramossima were 2.0 m and 3.0 m, respectively. The high cover P. euphratica distribution area was mainly concentrated in the near-banks of Donghe River and Xihe River, while higher cover T. ramossima was distributed in most parts of the study area. From the aspect of current groundwater level, T. ramossima should be the adaptive species in the study area. The similarities between the potential and actual spatial distribution of P. euphratica and T. ramossima were 43. 4% and 55. 6% , respectively, and the main reason for the lower similarity was that there existed a gypsum salt pan in soil, which blocked the vertical movement of soil water. PMID- 19288725 TI - [Assessment model of townlet eco-environmental quality based on BP-artificial neural network]. AB - Aiming at the problems in the townlet eco-environmental quality assessment in China, a comprehensive assessment model of townlet ecological environmental quality based on BP-artificial neural network was set up, which contained 20 cryptic layer nodes and 3 layers. The rank classification criterion of eco environmental quality's assessment indicator system were chosen as the training sample of the model, the 10% of training sample as well as the middle and critical values were regarded as examining sample, and the monitoring values of assessment region were treated as test sample. Choosing the Dushan Town in Ezhou City as an example, the training and prediction were made by using MATLAB software. The results showed that BP-artificial neural network was not only feasible and dependable, but also couldprovide acceptable results in accord with the regional realistic eco-environmental feature. PMID- 19288726 TI - [A new type water supplement mode of urban wetland park and its effects in purification and ecology]. AB - With the Rosebush Wetland Park in Changzhou as a case, a new type water supplement mode for urban wetland park, i.e., "vertical-flow plus horizontal flow", was constructed, and its effects in water purification, ecology, and economic advantages were evaluated. The results showed that this water supplement mode could not only improve the landscape of the water bodies in urban wetland park, but also enhance their quality, making it satisfy the requirement for human full-body exposure. Furthermore, the operation cost of the mode was as lower as 5%-25% of direct municipal pipe-water supply and other routine technique solutions, suggesting that this water supplement mode had potential positive ecological effects and economic advantages. PMID- 19288727 TI - [Microclimatic niche of rice sheath blight fungus]. AB - By using the principles and methods of niche, the microclimatic niche of rice sheath blight fungus was analyzed. The results showed that the integrative effect of ecological factors light intensity, air temperature, and humidity was higher than the effect of each of the factors, and the combination of light intensity and air temperature had higher effect than the other combinations of the three factors. At the tillering, booting, heading, and waxing stages of rice, the microclimatic niche breadth of the fungus was 0.3112, 0.4012, 0.4326, and 0.7365, respectively, i.e., increased with rice growth, and the microclimatic niche occupied by the fungus was mainly with the low light intensity, low air temperature and high humidity as the prior microclimate type. PMID- 19288728 TI - [Spatial distribution of arbuscular mycorrhizal fungi in Astragalus Adsurgens root-zone soil in Mu Us sand land]. AB - This paper studied the spatial distribution of arbuscular mycorrhizal (AM) fungi in Astragalus adsurgens root-zone soil in Mu Us sand land under five different ecological conditions. The results showed that the colonization and spore density of AM fungi differed significantly with sampling sites and soil depths. The highest colonization rate and spore density of AM fungi were found in 10-30 cm soil layer, and the highest spore density was at Dingbian site. Ningtiaoliang and Tawan sites had the highest vesicular colonization, and Tawan site had the highest total and hyphal colonization. The contents of total glomalin (TG) and easily extractable glomain (EEG) were the highest at Tawan site, being 1.18 mg x g(-1) and 0.65 mg x g(-1), respectively. Soil pH had significant positive effects on the vesicular and arbuscular colonization, spore density, TG, and EEG, and the TG and EEG had significant positive correlations with spore density, soil organic carbon (SOC), and soil available N and P. The proportion of glomalin to SOC was higher in desert soils than in agricultural soils, which implied that glomalin could be one of the main origins of SOC in desert ecosystem. Therefore, glomalin could be a useful index for the evaluation of soil AM fungal activity and soil ecology. PMID- 19288729 TI - [Effects of rotation and intercropping on bacterial communities in rhizosphere soil of cucumber]. AB - By the method of PCR-DGGE, this paper studied the effects of rotation with wheat, soybean, villose vetch, clover, and alfalfa and intercropping with onion and garlic on the bacterial communities in rhizosphere soil of cucumber. The results showed that rotation and intercropping with test plants increased the diversity and evenness indices of bacterial communities in cucumber rhizosphere soil, and also, cucumber yield. The sequencing of DGGE bands indicated that most of the bands had high homology with uncultured bacterial species, and were of Sphingobacterium and Proteobacteria. High bacterium G+C was only detected when cucumber was intercropped with onion. The diversity of soil bacterial communities varied with the growth stages of cucumber, being the highest at vigorous fruiting stage. It was suggested that intercropping with onion and rotation with wheat were the best cultivation modes of cucumber. PMID- 19288730 TI - [Arthropod communities in jujube-grain intercropping system: composition and time series dynamics]. AB - In order to reveal the correlations between the arthropod communities in two sub systems (jujube yard and grain field) of jujube-grain intercropping system, an investigation was conducted on the composition and time series dynamics of the communities in the two sub-systems. A total of 14 936 arthropod individuals belonging to 14 orders and 96 families were found, among which, 5992 individuals of 14 orders and 82 families were in jujube yard and 8971 of 14 orders and 80 families were in grain field, and some differences were observed in the arthropod community structure between the two sub-systems. The time series dynamics of the arthropod communities could be divided into four periods, according to the optimal cut-apart of time series, i.e., decline period in grain field sub-system, species number increase period in whole system, steady period in whole system, and decline period in whole system. The primary natural enemies in the system showed a characteristic of transferring between the two sub-systems in the four periods, and the parasitical natural enemies in a sub-system could impact the pest insects in another sub-system. The species transferring and mutual effect of the arthropod communities between the two sub-systems could benefit the stability of the jujube-grain intercropping system, and the capability of the system in controlling pest insects. PMID- 19288731 TI - [Comprehensive assessment on management measures of rice insect pests]. AB - Based on the investigation of arthropods biodiversity and insect pests controlling effect in the paddy fields of demonstration area of organic rice production in Guangzhou, a comprehensive assessment on the economic, social, and ecological profits and costs of organic rice production in study area was conducted by using analytic hierarchy process (AHP). The results indicated that among all insect pests controlling measures, organic rice planting had the best comprehensive benefits, with a weight value of 0.5355. Ecological cost played a dominant role in comprehensive costs, and that of traditional pesticide-based insect pests control was most cruel, with a weight value of comprehensive costs being 0.6252. From the viewpoint of the ratio of comprehensive profits to comprehensive costs (RPC), organic rice planting was better than any other insect pests controlling modes, whose RPC was 2.4776, being accorded with the requirement of sustainable development of agriculture. PMID- 19288732 TI - [Cadmium accumulation and its development tendency in black soil under long-term fertilization]. AB - Based on the long-term field experiment at Hailun Agro-ecosystem Experimental Station of Chinese Academy of Sciences, this paper studied the cadmium (Cd) accumulation, its development tendency, and Cd availability in black soil under effects of long-term application of chemical N and P fertilizers and pig manure. The results showed that under no fertilization, soil Cd concentration had a slight increase. Long-term chemical N and P fertilization increased soil Cd concentration significantly, but soil Cd accumulation had less difference under different N and P fertilization rates. Applying pig manure increased the Cd accumulation in soil significantly, and the accumulation had a tendency of speed up. Cd-containing feedstuff additives could be the important source of Cd in pig manure. No significant effects of chemical N and P fertilization were observed on the Cd availability in soil, but long-term application of pig manure increased the Cd availability significantly. PMID- 19288733 TI - [Effects of fungicide chlorothalonil on soil nitrous oxide and carbon dioxide emissions]. AB - A 14 d incubation test at 60% WHC and 25 degrees C was conducted to study the effects of fungicide chlorothalonil at its application rates of 0, 5.5 mg x kg( 1) (field application rate, FR), 110 mg x kg(-1) (20FR) and 220 mg x kg(-1) (40FR) on the nitrous oxide (N2O) and carbon dioxide (CO2) emissions from acidic, neutral, and alkaline soils. The results indicated that the effects of chlorothalonil on the two gases emissions depended on its application rate and soil type. Comparing with no chlorothalonil application, the chlorothalonil at 20FR and 40FR inhibited the N2O emission from acid soil significantly, while that at FR, 20FR and 40FR stimulated the N2O emission from neutral soil, with the strongest effect at FR. Higher application rates (20FR and 40FR) of chlorothalonil inhibited the N2O emission from alkaline soil at the early stage of incubation, but stimulated it at late incubation stage. Chlorothalonil at FR had no obvious effects on the CO2 emission from test soils, but that at 20FR and 40FR promoted the CO2 emission from acid soil while inhibited it from neutral and alkaline soils significantly. PMID- 19288734 TI - [Risk assessment of pig manure Cu-contamination of black soil in northeast China]. AB - Copper (Cu) is massively used as feed additives in intensive farms, and the discharge and utilization of livestock wastes may cause certain environmental problems. In this paper, a cultivated black soil in Northeast China was sampled, and added with pig manures that contained different concentration of Cu to simulate the vegetable soil under different years of pig manure fertilization. With this soil, a pot experiment was conducted to study the effects of soil Cu accumulation via the application of the pig manures on the aboveground biomass and its Cu concentration of pakchoi (Brassica chinensis L.), soil microbial biomass carbon, and soil enzymes (dehydrogenase, urease, and acid phosphatase) activities. The results showed that pig manure application increased the aboveground biomass of pakchoi significantly, but had no significant effects on the aboveground biomass Cu concentration. Under the application of pig manure, soil dehydrogenase and urease activities increased in a definite period of time, but their activities as well as soil acid phosphatase activity were depressed gradually with the increase of soil Cu concentration. When the soil total Cu concentration reached 301.3 mg x kg(-1), the aboveground biomass of pakchoi, soil microbial biomass carbon, and the activities of dehydrogenase, urease, and acid phosphatase were greatly reducd. The aboveground biomass Cu concentration of pakchoi had significant positive correlations with the concentration of soil total and water soluble Cu (P < 0.01), while the aboveground biomass was negatively correlated with soil total Cu concentration (P < 0.05). It was suggested that the threshold value of Cu in black soil under vegetable cultivation should be less than 301.3 mg x kg(-1). PMID- 19288735 TI - [Research advances in cadmium pollution of peanut (Arachis hypogaea L.)]. AB - Peanut (Arachis hypogaea L.) is a major oil-bearing crop in the world, and as well, an important resource of plant protein and a main raw material for food processing. With the increasing of its direct human consumption and food processing, the Cd concentration in peanut kernel has aroused great concern in recent years. China is a main country of the production and exportation of peanut, but the Cd enrichment in peanut kernel is the main obstacle for its peanut export trade. In this paper, the research advances in Cd pollution of peanut kernel were reviewed, based on the characteristics and mechanisms of Cd accumulation and distribution in peanut kernel, the intra-specific variation of kernel Cd content, and the measures in controlling kernel Cd content. Two strategies were put forward for controlling Cd pollution of peanut kernel, i.e., to reduce the Cd uptake by main root system of peanut plant, and to control the transference of Cd from root to fruit (kernel). In order to applying the strategies effectively, researches on the mechanisms of Cd accumulation in peanut kernel should be enhanced in three aspects, i.e., root vitality and its relationship with Cd accumulation in kernel, mechanism of fruit Cd absorption and its contribution to kernel Cd content, and mechanism of Cd transference in plants and its effects on kernel Cd content. PMID- 19288736 TI - [Form transformation of arsenic in soil and corresponding analyzing methods]. AB - Based on the analysis of the sources and existing forms of soil arsenic, this paper approached the inter-transformationr of different arsenic forms in soil. In the meanwhile, the extraction and determination methods of different soil arsenic forms were also compared. It was considered that HPLC-HG-AFS had the advantages of high sensitivity, low detection limit, better selectivity, low operation cost, and less inter-transformation of different arsenic forms, being able to be used as the prior method for the detection of different arsenic forms. Combining with previous research results, the form transformation of arsenic in soil and the promising research aspects were also discussed. PMID- 19288737 TI - [Soil enzyme activities under two forest types as affected by different levels of nitrogen deposition]. AB - A simulation test was conducted to study the change trends of soil cellulase, polyphenol oxidase, and sucrase activities under natural broadleaf-Korean pine (Pinus koraiensis) and secondary poplar (Populus davidiana) -birch (Betula platyphylla) mixed forests as affected by 0, 25, and 50 kg x hm(-2) x a(-1) of N deposition. The results showed that the effects of elevated N deposition on test enzyme activities varied with forest type, and short-term nitrogen addition could significantly affect the test enzyme activities. High N deposition decreased soil polyphyneol oxidase activity, and correspondingly, soil cellulase and sucrase activities also had a trend of decrease. PMID- 19288738 TI - [Biological process of phosphorus turnover in surface water body of Xiamen Harbor. II: Grazing pressure of copepod on phytoplankton]. AB - To understand the roles of copepod in the biogeochemical cycling of phosphorus, gut fluorescence method was applied to examine in situ the grazing rate of copepod on the phytoplankton in Xiamen Time Station (XMTS) in May, August and November 2005 and March 2006. In the meanwhile, the abundance and species composition of copepod were investigated, and the grazing pressure of copepod on the phytoplankton was estimated. The results showed that the annual average grazing rate of copepod was 55.53 microg x m(-3) x d(-1), being the highest (108.98 microg x m(-3) x d(-1)) in autumn and the lowest (7.18 microg x m(-3) x d(-1)) in summer. Based on the estimation from our experimental data, the daily grazing rate of copepod populations on the phytoplankton in Xiamen Harbor was, on annual average, about 1.81% of the phytoplankton's standing stock, with the values in spring, summer, autumn, and winter being 3.22%, 0.06%, 3.52% and 0.46%, respectively. PMID- 19288739 TI - [A decade's evidence review of sildenafil citrate]. AB - Erectile dysfunction (ED) is one of the most popular and disturbing sexual problems, which impairs patients' health and affects their quality of life. Sildenafil citrate, the first oral PDE-5 inhibitor in the world, has established its efficacy and safety credit in both doctors and ED patients. The authors reviewed the results of related studies in the last decade and made deeper insights into the use of sildenafil citrate in such aspects as the general drug tolerance and its influence on the cardiovascular system, potentiality of concomitant diseases with co-administration of other drugs, long-term safety, and ocular and genital safety. PMID- 19288740 TI - [Correlation of prostate cancer susceptibility with genetic polymorphism of cytochrome P450 2E1, smoking and drinking: a case-control study in the population of Nanjing area]. AB - OBJECTIVE: To investigate the association of the risk of prostate cancer (PCa) with the polymorphism of the CYP2E1 gene, smoking and drinking, and to explore the joint role of genes and living habits in PCa pathogenesis. METHODS: We conducted a case-control study on 109 PCa patients and 202 age-matched non-PCa male controls, and detected the polymorphisms of CYP2E1 Rsa I and Pst I sites by PCR-RFLP using DNA from peripheral blood lymphocytes. RESULTS: The history of deep smoking (OR = 2.29, 95% CI: 1.28 - 4.09) or heavy smoking (OR = 1.81, 95% CI: 1.02 - 3.22) was a risk factor. The CYP2E1 C1/C1 genotype significantly increased the risk of PCa (OR = 1.71, 95% CI: 1.04 - 2.82) and apparently interacted with drinking (OR = 2.21, 95% CI: 1.06 - 4.59). Heavy smokers with the C1/C1 genotype showed an increased risk of PCa (OR = 2.80, 95% CI: 1.20 - 6.56), as compared with non-smokers carrying the genotype of C1/C2 or C2/C2. CONCLUSION: The risk of PCa obviously increases in individuals with both the CYP2E1 C1/C1 genotype and the habit of smoking or drinking, and it has a significant positive correlation with the dose of tobacco exposure. PMID- 19288741 TI - [Duckett's procedure of one-stage urethroplasty for hypospadias]. AB - OBJECTIVE: To study the clinical effect of Duckett's procedure of one-stage urethroplasty with the transverse preputial island flap in the treatment of hypospadias. METHODS: We retrospectively studied 20 cases of hypospadias treated by Duckett's procedure of one-stage urethroplasty with the transverse preputial island flap (Group A) from 2003 to 2007. The patients ranged in age from 2 to 22 years (mean 11.3 years). Another 30 hypospadias patients aged 7 to 34 (mean 16.1) years underwent urethroplasty with mucous membrane of the bladder (Group B) in the same period. Of the 50 cases, 13 were the penile type, 33 the penoscrotal type, 2 the scrotal type, and 2 the perineal type. Catheters were indwelled in Group A, but both cystostomy and catheter indwelling were used in Group B. RESULTS: For Duckett's procedure, surgery succeeded in 17 cases (85%), urethra stenosis developed in 1 (5%) and urinary fistula in 2 (10%), which were successfully repaired 6 months after the operation. The mean operation frequency was 1.1 per patient. In Group B, the operation was accomplished in 2 stages and succeeded in 26 cases (86.7%), with postoperative development of urinary fistula in 3 (10%) and urethra stenosis in 1 (3.3%). The mean operation frequency was 2.1 per patient. The 2 cases of postoperative urethra stenosis were both cured by urethral sounding. CONCLUSION: Duckett's procedure of one-stage urethroplasty with the transverse preputial island flap is effective in hypospadias repair, with a success rate close to that of other types of operations and a low rate of postoperative complications. Compared with the use of mucous membrane of the bladder, it has the advantages of one-move accomplishment and gives a better penile appearance. PMID- 19288742 TI - [Sperm acrosin activity helps predict IVF-ET outcome]. AB - OBJECTIVE: To investigate the effect of sperm acrosin activity on the IVF-ET outcome. METHODS: We analyzed sperm parameters, morphology and acrosin activity for 909 infertile husbands by computer-assisted self-assessment (CASA), modified Papanicolaou staining and N-alpha-benzoyl-DL-arginine-p-nitroanilide (BAPNA), respectively, and detected the rates of fertilization, cleavage, quality embryos, embryo cryopreservation, implantation, clinical pregnancy and abortion. The wives were identified as normal or with mere oviduct problems. RESULTS: The rate of normal sperm morphology and sperm motility, vitality, rapid progressive velocity and concentration were significantly lower in the abnormal acrosin activity group than in the normal one (P < 0.01). Significant positive correlations were observed between acrosin activity and the above-mentioned semen parameters (P < 0.01). There were no significant differences in the number of retrieved eggs, the rates of cleavage, quality embryos, embryo cryopreservation, non-embryo transfer cycles and miscarriages, and the number of transferred embryos between the two groups (P > 0.05). The fertilization rate, the percentage of transfer cycles with only 1 embryo and the rate of implantation and clinical pregnancy were notably higher in the normal acrosin activity group than in the abnormal one (P < 0.01). CONCLUSION: Sperm acrosin activity is closely related with semen parameters, and it helps to predict the sperm fertilizing capacity and IVF-ET outcome. PMID- 19288743 TI - [Identification of M(r)22 000 protein in rat epididymal luminal fluid]. AB - OBJECTIVE: To identify the specific protein in the epididymal luminal fluid that may play a role in sperm epididymal maturation or modification on the surface of spermatozoa. METHODS: We compared the differential protein components in the lumen fluids from the caput and cauda segments of the epididymis of normal rats as well as from the cauda segment of experimental left varicocele (ELV) rats by SDS-PAGE or 2D-electrophoresis. The protein spots of interest were selected for MS identification, and the target proteins further characterized by immuno-blot assay. RESULTS: MS analysis showed that one of the most prominent proteins, M(r) 22 000, was identical to the phosphatidylethanolamine-binding protein (PBP), and it was further identified as PBP by immuno-blot assay. CONCLUSION: PBPs were present in a variety of molecular forms in the epididymal luminal fluid, including the glycosylated form, and ELV markedly elevated the PBP level in the cauda luminal fluid of the rats. Thus, the association of this molecule with sperm surface modification remains an interest for future investigation. PMID- 19288744 TI - [Expressions of STAT3, VEGF and C-myc in seminoma and their significance]. AB - OBJECTIVE: To understand the role of the JAK/STAT3 signal transduction pathway in the pathogenesis of seminoma by studying the expressions of signal transducers and activators of transcription-3 (STAT3), vascular endothelial growth factor (VEGF) and C-myc in seminoma. METHODS: We examined 38 paraffin specimens of seminoma and 10 samples of normal human testes by immunohistochemical staining using the antibodies of STAT3, VEGF and C-myc, and observed the staining intensity under the light microscope. RESULTS: The positive expression rates of STAT3, VEGF and C-myc were 76.3%, 71.1% and 84.2%, respectively, with statistically significant differences from the corresponding protein expressions in the normal testis tissues (P < 0.01), which gradually increased with the clinical stages of tumor, nodes and metastasis (TNM) classification. Different correlations were observed among the STAT3, VEGF and C-myc proteins in seminoma: STAT3 positively correlated with VEGF (r = 0.640, P < 0.01) and C-myc (r = 0.408, P < 0.05); C-myc positively correlated with VEGF (r = 0.459, P < 0.01). CONCLUSION: The JAK/STAT3 signal transduction pathway can facilitate the development and metastasis of seminoma by activating the expression of VEGF and meanwhile induce the malignant proliferation of primary reproductive cells and promote the pathogenesis and progression of seminoma by activating the expression of C-myc. PMID- 19288745 TI - [Finasteride: an effective therapeutic for benign prostatic hyperplasia related gross hematuria in patients receiving anticoagulant]. AB - OBJECTIVE: To study the effect of finasteride on benign prostatic hyperplasia (BPH) related gross hematuria in patients receiving anticoagulant. METHODS: A total of 105 patients with BPH related gross hematuria were divided into an anticoagulant group (n = 81), treated with combined therapy of anticoagulants and finasteride, and a control group (n = 24), given finasteride only at 5 mg daily. The therapeutic effects were compared by a 6-month follow-up. RESULTS: In the anticoagulant group, gross hematuria was cured in 52 patients (64.2%), taking an average time of 3.9 weeks (1-6 weeks), and improved in 12 patients (14.8%), as compared with 16 patients cured (66.7%), 3.2 weeks taken (1-5 weeks), and 4 patients improved (16.7%) in the control group. The mean time taken to resolve hematuria was longer in the former (P < 0.05). But the cure rates had no significant differences either between the two groups or among the subgroups receiving different anticoagulants. CONCLUSION: Finasteride is an effective therapeutic for BPH related hematuria in patients receiving different anticoagulants. It makes no significant differences in cure and effectiveness rates between patients treated with and without anticoagulant, but takes an average of longer time to resolve hematuria in patients receiving anticoagulant. PMID- 19288746 TI - [Recurrent extramammary Paget's disease of the penis and scrotum: clinical analysis of 18 cases]. AB - OBJECTIVE: To investigate the causes, therapeutic methods and prognosis of recurrent extramammary Paget's disease of the penis and scrotum. METHODS: We retrospectively studied the clinical presentations, therapeutic methods and prognosis of 18 cases of recurrent penile and scrotal Paget's disease, all diagnosed by biopsy and treated by surgical resection of the diseased sites. The incision reached the deep fascia. Frozen and olefin sections of the samples were pathologically examined to identify the involvement of the incisal edge. Seven cases had inguinal lymph node metastasis and underwent unilateral or bilateral inguinal lymph node resection. RESULTS: All the patients were followed up for 6 months to 8 years (mean 34 months). Four died of distant metastasis and the other 14 experienced no recurrence. CONCLUSION: For the cases without metastasis of recurrent penile and scrotal Paget's disease, extended lesion-resection is a preferred option and the prognosis is good. And those with local lymph node metastasis need to undergo local lymph node resection. PMID- 19288747 TI - [Effect of thyroid hormone on the contents of NOS and CO in the penile corpus cavernosum of rats]. AB - OBJECTIVE: To determine the contents of NOS and endogenous CO in the penile corpus cavernosum of Wistar rat models of hyperthyroid and hypothyroid, to detect the effects of thyroid hormone and endogenous CO on rat penile erection, and to further investigate the influence of thyroid hormone on human penile erection. METHODS: Fifty 3-month-old male Wistar rats were randomly and equally divided into 5 groups: a hyperthyroid, a hyperthyroid treatment, a hypothyroid, a hypothyroid treatment and a control group. The contents of NOS and CO were detected by ultraviolet spectrophotometry. RESULTS: Compared with the controls, the content of NOS was significantly decreased in both the hyperthyroid and the hypothyroid groups (P < 0.01), even lower in the latter than in the former (P < 0.01), and so was the content of CO (P < 0.01), lower in the former than in the latter (P < 0.01). After treatment, the levels of NOS and CO were both elevated in either the hyperthyroid or the hypothyroid group, with no significant difference from that of the controls (P > 0.05). CONCLUSION: The contents of NOS and CO in the penile corpus cavernosum decrease with the imbalance of thyroid hormones, but return to normal once the imbalance is corrected. Under the same conditions, hypothyroid inflicts more damage on penile erection than hyperthyroid. PMID- 19288748 TI - [Expression of Smad4 in prostate cancer LNCaP and ARCaP cell lines with different metastatic potentials and its significance]. AB - OBJECTIVE: To observe the expression of Smad4, the core of TGF-beta/Smads signal transduction pathway in different prostate cancer cell lines, and explore their molecular mechanism of bone metastatic potential. METHODS: The Millicell polycarbonate filter coated with matrigel was used to confirm the invasive potency of LNCaP and ARCaP cell lines (IF11 and IA8). The expressions of the Smad4 protein and mRNA in these prostate cancer cells with different metastatic potentials were detected by Western blotting and RT-PCR, respectively. RESULTS: ARCaP cell lines (IF11 and IA8) exhibited a stronger potency of invasion than LNCaP (P < 0.01). The Smad4 protein and mRNA highly expressed in the LNCaP cell line that was well-known with a low metastatic potential, but not in the ARCaP (IF11 or IA8) cells with high metastatic potentials (P < 0.01). CONCLUSION: Smad4 expresses differently in LNCaP and ARCaP cell lines with different metastatic potentials and, as a tumor suppressive gene, its deficient expression may play an important role in the invasion and metastasis of advanced prostate cancer. PMID- 19288749 TI - [Replacement of plaque by buccal mucosa in the treatment of Peyronies disease: a report of 27 cases]. AB - OBJECTIVE: To evaluate the clinical results of the replacement of plaque by buccal mucosa in the treatment of Peyronie's disease. METHODS: We performed surgical replacement of the plaque by free autograft of buccal mucosa on 27 patients with Peyronie's disease, ranging in age from 24 to 72 years (mean 53), varying in disease course between 1 and 13 years, with a penile curvature angle of 30 degrees - 80 degrees in erection. Of the total number, 15 patients had the plaque located in the dorsal root, 6 in the dorsal shaft, 3 in the subcoronal shaft and 3 in the ventral shaft of the penis; 24 had 1 plaque, 2 had 2 and 1 had 3, ranging in size from 0.7 cm x 0.7 cm to 1.6 cm x 1.0 cm. RESULTS: Satisfactory results were achieved in all the cases, with no such complications as hematoma, infection, oral numbness, and tightness of the mouth. Of the 24 cases that were followed up for 0.5 to 7 years, complete straightening of the penis was achieved in 21, slight residual curvature (< 15 degrees) was noted in 3, a little shortening of the penis (< 1 cm) in 2, and erectile pain in 3. CONCLUSION: Buccal mucosa, with a stable elasticity and no shrinkage, can be recommended for wide use in the surgical treatment of Peyronie's disease. PMID- 19288750 TI - [Clinical analysis and management of disappointing early urination after TURP]. AB - OBJECTIVE: To explore the etiology and management of disappointing early urination in patients with benign prostatic hyperplasia (BPH) following transurethral resection of the prostate (TURP). METHODS: The clinical data and management of 58 cases of disappointing urination within 3 months after TURP were reviewed. RESULTS: Of the total number of patients with disappointing early urination after TURP, 7 (12%) underwent a second operation, 6 (10%) received outpatient treatment, and the other 45 (78%) showed spontaneous improvement within 1 year with no further treatment. CONCLUSION: The causes of disappointing early urination after TURP included bladder neck contracture, urethral stricture and urinary tract infection, which necessitate secondary treatment. And a greater proportion of the patients showed functional lability of the bladder detrusor muscle, which could be spontaneously improved within 1 year after TURP. PMID- 19288751 TI - [Stra8: a specifically expressed gene in premeiotic germ cells' transition from mitosis to meiosis]. AB - Stra8 (stimulated by retinoic acid gene 8) is a specific expression gene in mammalian germ cells' transition from mitosis to meiosis. Stra8 expresses in an anterior to posterior wave from embryonic days 12.5 to 16.5 in germ cells of XX gonads in mice. Meiosis germ cells are observed 1 day after Stra8 expression, which begins in the testis after birth and triggers meiosis entry. Germ cell sex determination is directed by a difference in the timing of entry into meiosis in embryonic gonads. The position of retinoic acid production and CYP26b1 expression that metabolizes RA to an inactive form regulates Stra8 expression. Although cytoplasmic protein Stra8 is necessary for the entry of germ cells into meiotic prophase, the function of the protein remains unknown. PMID- 19288752 TI - [Multiple sclerosis and erectile dysfunction]. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the human central nervous system. NO, ion channel, cytokine and testosterone play an important role in MS, and may be associated with the pathogenesis of ED. Meanwhile, the relationship between MS-induced peripheral nerve injury and ED should be understood correctly. Further researches on these mediators can provide some theoretical evidence for the clinical treatment of ED. PMID- 19288753 TI - [Status quo of the researches on HSV vaccines]. AB - Herpes simplex virus (HSV) infection can cause severe recurrent disease in humans and establish lifelong latency in the host. Furthermore, it can significantly increase the risk of HIV infection and bring substantial psychosexual as well as physical morbidity to the patients. Several antiviral therapies are available for the control of its symptoms and spreading, but they can neither cure nor alter the nature of HSV infection. The development of an efficacious HSV vaccine is therefore necessitated for controlling the occurrence, transmission and recurrence of the infection. Those that have undergone clinical evaluation include subunit vaccines, attenuated live virus vaccines, replication defective virus vaccine, naked DNA vaccines and viral vector vaccines. Most of the above vaccine strategies have elicited protective immunity in animal models, but none has yet been effective in human beings. For all that, scholars have never stopped their exploration for an effective vaccine against the notorious virus. The authors present an overview of a few most promising candidate vaccines for the prevention and treatment of HSV infection. PMID- 19288754 TI - [Reconstruction and repair of resected cavernous nerves]. AB - By studying the novel methods for reconstructing damaged cavernous nerves and the related literature on the regeneration of cavernous nerves, restoration of erectile function and neurohistological reconstruction engineering, a variety of grafting materials have been found applicable to cavernous nerve reconstruction, including autogenetic nerve grafts, silicone tubes, artificial biodegradable conduits and so on. Neurotrophic factors, extra cellular matrix components and Schwann cells have been shown to promote cavernous regeneration. Artificial nerve guides, especially biodegradable ones containing growth-promoting factors or cells, are a promising option for the repair of cavernous nerve lesions. PMID- 19288755 TI - [Qiantongding Decoction: An efficacious therapeutic for type III B prostatitis]. AB - OBJECTIVE: To evaluate the clinical efficacy of Qiantongding Decoction in the treatment of type III B prostatitis or chronic pelvic pain syndrome (CPPS). METHODS: Seventy patients with type III B prostatitis / CPPS were randomly divided into an experimental group (n = 36) and a control group (n = 34), the former treated with Qiantongding Decoction, and the latter with oral Indometacin, both for a month. Then their scores on Pelvic Pain Symptom Survey and NIH-CPSI were documented and compared. RESULTS: An obvious improvement was observed in both groups after the treatment, with significant differences from pre-treatment (P < 0.05), and the experimental group showed a significantly better alleviation of pain than the control (P < 0.05). CONCLUSION: Qiantongding Decoction has an obvious therapeutic effect on type III B prostatitis / CPPS. PMID- 19288756 TI - [Tadalafil for erectile dysfunction: excellent safety and tolerance]. AB - As a novel phosphodiesterase type 5 (PDE5) inhibitor, tadalafil is administered orally for the treatment of erectile dysfunction (ED). In addition to its outstanding efficacy that lasts 36 hours, tadalafil shows an excellent safety and tolerance, with only a few transient mild to moderate adverse events. Usually ED patients are concomitantly afflicted with multiple pre-existing medical conditions or receive a range of drugs at the same time. Therefore, it is essential to understand its safety in general population and some special high risk cohorts. PMID- 19288759 TI - 'About Helen'--antipsychotic medication use during pregnancy. A case study. PMID- 19288760 TI - Nurse education. Our students. Our future. PMID- 19288761 TI - Working life. PMID- 19288762 TI - Need for stronger laws to protect older Australians. PMID- 19288763 TI - Preventing heat stroke in Australian communities. AB - This clinical update has identified the physiological aspects of heat acquisition and its effect on the human body. It has been shown that both classical (non exertional) and exertional heat stroke are preventable medical emergencies. Through a greater understanding of the physiological aspects of heat acquisition and the effects of that burden on the human body it has been shown that preventative strategies can be developed to reduce the prevalence of heat stroke. These simple measures can be applied to combat the detrimental effects of exposure to natural heat in the Australian community and prevent deaths. PMID- 19288764 TI - Solving the puzzle. Caring for older people in ED. PMID- 19288765 TI - Increased fire death risk for the elderly. PMID- 19288766 TI - Diabetes epidemic: implications for nursing practice. PMID- 19288767 TI - Diabetes in aged care. PMID- 19288768 TI - Sticking together in tough times. PMID- 19288769 TI - Caregiver--what help for the helpers? PMID- 19288770 TI - [Bioethics and caregiving: a critical interaction in patient care today]. AB - This paper outlines the main instruments and contexts of applied clinical ethics in which the ethical issues of caregiving and the figure and role of the caregiver, far from being in apparent conflict, complement and influence each other reciprocally: i.e. Informed Consent and the end of life decision making processes regarding the activation and suspension of vital support means. These in fact are the two principal areas, though there are other scenarios in which the correspondence between bioethics and caregiving would equally come to assume great importance (e.g. the doctor-patient-caregiver relationship, palliative care, the approach to suffering, terminal sedation, home care and treatments). Attention is drawn in particular to one end of life issue that is often overlooked, i.e. favoring the caregiver's presence as the time of dying draws near and at the moment of death itself, a presence that involves high emotional involvement and has a high moral impact and that can give dignity and spirituality to an event that patients usually experience in solitude and in the absence of family members and health professionals. The paper, in conclusion, highlights that in modern scenarios of the ethics of caregiving, particularly regarding the care of patients with chronic progressive diseases with negative prognosis or of patients with limited or no decisional capacity, the role of the caregiver is fundamental both as an element of balance between autonomy and beneficence and as a guarantor of the patient's best interest and of their preferences, desires and values, not to mention the contribution caregivers make to the ethical discernment and sense of responsibility of health professionals. PMID- 19288771 TI - [The caregiver of a person with Alzheimer's disease]. AB - The caregiver of a person with Alzheimer's disease is most likely to be a woman (in almost 80% of the cases); it may be a daughter, the wife, a sister, a daughter-in-law. The burden of this role is mainly practical (6 hours per day of direct assistance and 7 of surveillance; among the caregivers still working, almost 50% refer changes/worsening in his/her professional life; more than 20% of caregivers refer an evident impact on their health situation; the average cost per patient, including both family costs and society costs, is slightly more than 60,000 euro; but this cure occupation has, on caregiver's life, consequences that can be considered harmful for his or her relationships. For the caregiver, psychological help is fundamental to tackle emotions and difficulties in relating with other people. Each person tackles this occupation in a different way; this is due to the fact that each family has its own unique history and own unique way to relate to other people. On the whole, we can see that these responsibilities are taken only by the families and in particular by the caregiver, great social and economical charges. A redistribution of the resources to help reduce the expenses of the families; more flexible institutional and organizational models; ethical laws that help defend the patients and the caregivers rights, would all certainly make our country a more civilized one. PMID- 19288772 TI - [Perception and opinions on the right of choice in medical field: what caregivers teach us on the communication]. AB - The aim of the present study was to analyse caregivers' perceptions and opinions concerning medical information and advance directives. Fifty-one subjects (mean age 63.07+/-12.2) were enrolled and filled in some questionnaires: the Family Strain Questionnaire (FSQ), the SF-12 and a specifically assembled questionnaire which we named Communication and Consent (CCQ). Correlations were found between FSQ factors, which refer to caregiving-related problems, and SF-12, which refers to the self-perception of mental and physical health. The subjects who resulted emotionally distressed referred to care for more critical patients and that they had not received sufficient attention by physician at the time of diagnosis. Self determination and decision-making resulted inversely correlated to caregivers' age and education. In general, caregiver's role itself influences attitudes towards medical information and advanced directives. The study also indicates that health professionals have to pay more attention to the relationship than they use to do, trying to understand which kind of emotional sound a technical term may produce for the patients and their caregivers. PMID- 19288773 TI - [Can the new technologies of telemedicine applied to health help the caregiver?]. AB - During the last few years about the chronic patient assistance the tendency is to privilege the home care model, favouring the permanence of the patient in the familiar nucleus. This determines an always greater involvement in term of time and responsibility of the caregiver that is of the person who takes cure of the patient one worrying itself to answer to its physical needs, psychical and social. The burden of the family caregiver is in the consisting majority of the cases rather. The caregiver is therefore, with full rights, the other protagonist of the disease and it must be necessarily integrated in the assistance plan. The increase of the age associated to an increase of the prevalence of chronic pathologies, determines the necessity to plan new interventions on the territory. In chronic patients alternative assistance models, using telemedicine, seem to be effectives improving both clinical aspects and quality of the life. A new area of interest is delineated therefore that, through the new technologies of the ICT must define been involved the single roles of the operating ones in the participation program. The telemedicine seems to be a useful instrument in order to support patient and caregiver in facing the disease and reducing stress. In our model of domiciliary telesurveillance the patient, the caregiver, the family and all the sanitary figures are been involved. This model integrating the service dedicated to chronic pathology with telepsychology at home seems to give good result even if ulterior studies, above all in the long term, are need. PMID- 19288774 TI - [The caregiver in oncology: duties and needs]. AB - Cancer is not only an individual problem, but it affects the entire family, disrupting relationships, daily routines and social relations. In our country, assistance to cancer patients is mostly the responsibility of the family and in particular, of those who are in charge with administering the treatment: the caregivers. Based upon scientific research, the present paper is aimed at focusing attention to caregivers, going through their responsibilities during the different stages of the disease and the possible outcomes on their well-being and quality of life. The caregiver meets the patient's requirements mainly in two respects: medical nursing care and emotional well-being. Research demonstrated the outcomes on the psychological and physical balances and on the daily activities of the family as well as the professional and social areas. The caregiver has a variety of practical and emotional needs. In particular one need goes throughout all the stages of the disease: a clear and correct communication, that could improve the relationships between patient, caregiver and physician, providing exhaustive and comprehensible information about all the aspects of the disease. The caregiver, in fact, takes on responsibilities that are physically and emotionally demanding, considering that he/she does not have any specialisation for this role. On this basis, more information is needed about the disease and more psychological support, with special reference to emotional aspects. PMID- 19288775 TI - [The family context in end-of-life is a resource to know and support: some reflections]. AB - The family caregiver role is a vital element of home-based patient care. At home, the achievement of the care plan may depend on the physician's relationship with the patient's caregiving network. Caregivers often tolerate an incalculable emotional burden for their work. Although many enjoy a profound sense of benefit and get deep satisfaction in this role, sadness, guilt, anger, resentment, and a sense of inadequacy are also common and understandable reactions. Exhaustion, financial strain, disrupted usual activities, and continuous caregiving contribute to significant mental health morbidity, including anxiety and depression. Early recognition of caregiver distress, help to secure a trusting partnership between the physician and the patient-caregiver unit at the end of life. This paper give some suggestion coming from a decade of support to families during the end stage. PMID- 19288776 TI - [Prolonged grief disorder: towards a new diagnostic category]. AB - The grief in response to loss of a significant is a normal, inevitable, experience of life. Nevertheless some people, even after a couple of months, do not succeed in integrating this experience in their daily life and remain stuck in a state of suffering condition that seriously extended in the time is functionally impairing. For long time psychology has focused on bereavement subsequent to the loss and the complications that can derive from the missing elaboration. The condition of prolonged grief has been object of a large debate in the past years, up to the individualization of a specific set of symptoms identified in a new diagnostic category: the Prolonged Grief Disorder (PGD). The PGD is featured as a condition of emotional distress and physical distress by the loss, not for the death, of a significant only, with daily involvement in all function areas. Our work given a short and non exhaustive overview of the cultural and scientific run that has brought to the collection of the evidences to insert the PGD as new diagnostic category in the DSM V and a comparison between the Post-Traumatic Stress Disorder (PTSD) and the Major Depressive Disorder (MDD) for better underlining similes, but above all differences among categories that subtend clinical conditions that can appear sometimes overlaps. PMID- 19288777 TI - An Internet-based cognitive-behavioral preventive intervention for complicated grief: a pilot study. AB - Bereaved parents and individuals bereaved by the sudden, unexpected, or violent death of a loved one are at high risk for developing complicated grief. However, preventive interventions for this at-risk population remain scarce. In the present pilot study, an Internet-based intervention for individuals bereaved in the previous 14 months was tested using a pretest/posttest design. A total of 35 bereaved individuals participated in a 3-week manualized prevention program. Counselor and participants communicated exclusively via email. The intervention consisted of the following modules: (1) describing the circumstances of the death, (2) using the life-imprint method to explore the biography and life imprint of the deceased, (3) keeping a daily diary of social activities and sleep hygiene, (4) cognitive restructuring of dysfunctional thoughts, such as responsibility for the death and feelings of guilt, (5) communication within the family, (6) gender-specific coping with bereavement, and (7) bond with the deceased. Completer and intent-to-treat analyses suggest that the intervention led to a significant reduction in symptoms of complicated grief and depression at posttreatment. PMID- 19288778 TI - [Chronic disease impact on partners. An explorative study]. AB - In the last years there has been a growing interest of researchers for themes dealing with chronic illness and issues related to that condition. Several authors have studied the difficulties caregivers have to cope with, especially when the patient's partner has this role. That condition would have both physical and psychological consequences, but its real effects seem to depend on a complex set of interacting elements. On the basis of these researches, in the current study we aimed to explore the influence of adult attachment and marital satisfaction on the impact that chronic illness can have on patient's partner and on his/her health-related quality of life. The sample is composed of 40 chronic renal failure patients' partner-caregivers. Each caregiver filled in a battery of tests for finding out the characteristics of adult attachment and marital satisfaction, health-related quality of life and the association between these variables and the impact of chronic illness on partners. Descriptive statistics, correlations and one-sample t test analysis have been performed. Results show a significant correlation between attachment style and caregiving burden; there are no gender differences in disease impact on caregiver. Moreover caregivers seem to have a worse quality of life in comparison to general population. Understanding caregiving can lead, at a future time, to planning specific interventions for both patients and partners, using a global and integrated approach that respects individuals and couples in their biologic, psychological and relational dimensions. PMID- 19288779 TI - [Working with families in the early stages of psychosis: a structured intervention for caregivers]. AB - In the field of early psychosis psychoeducation is considered fundamental to increase coping skills with diseases and to improve the quality of life of patients and their families. The more recent and updated guidelines on schizophrenia underline the extreme importance of the families involvement in treatment of young people in the initial phases of illness. "Families are the main support for many young patients. They could be the primary carers but they have also to face individual and social consequences following the onset course. Where feasible, family members must be involved in the treatment". This work describes the components of the work with families carried on by the Centre for the early detection of psychoses and high-risk situations--Programma 2000 ("Niguarda Ca' Granda" Hospital-Milan) and is mostly focused on psychoeducation and on Expressed Emotions aspects. Even the advances suggested by the international literature drove Programma 2000 to define both the steps of caregivers assessment and intervention. During the last ten years, Programma 2000 has followed 191 caregivers. Aims of this work is to verifier the outcome of the "pilot project", started in 2007, projected specifically to increase the normally used strategies to improve the caregivers adherence and involvement in the therapeutic process. The individualized multi-componential intervention has been structured in 8 session over one years. Outcome measures used in this article are the scores of the Camberwell Family Interview and from the Psychosis Knowledge Assessement Semistructured Interview (VCP). The subjects enrolled in the structured pilot project were 25 family caregiver to young (18-30 yrs old) patients. Results shows change in the Expressed Emotion level: 13% of families moved from High Expressed Emotion to Low Expressed Emotion. Furthermore data on the knowledge of illness knowledge level illustrate a reduction in the percentage, from 47% to 18%, of carers who have just a very vague knowledge of illness, and an increase from 16% to 27% of carers who obtain a good level of specific knowledge. In conclusion we can sustain mental health expert with aim to treatment project programme individualized and multi-componential tailored for young's caregiver at the onset phase of psychosis. PMID- 19288780 TI - [Therapeutic relationship with the child and the adolescent affected with leukemia]. AB - The main psychosocial purpose in treating childhood cancer is to help children and their families to face the diagnosis of cancer and subsequent consequences. Children and their families, most of whose are in front of this new diagnosis without showing any sign of failure, need our help. We should try from one side to help the child and his/her family who need a very quick support from us, from the other side a controlled and scientifically valid research, finalized to differentiate effective from non-effective interventions, should be carried on. The optimal clinical assistance is related to the application of the best discoveries nowadays available, based on evidence and applied in the local cultural context. The health care team can carefully listen to the children and their families to detect in which way they work and answer to the request of assistance that was offered to them. To modify own approach based on the level of satisfaction of families looking at the type of offered assistance could help in making better service. PMID- 19288781 TI - [Adoptive parental couple: a pilot study]. AB - International adoption is a phenomenon in constant growth, involving over one hundred countries. In Italy there are approximately two thousand adoptions of foreign minors per year. The needs demonstrated by the families adopting these children, sometimes bearing complex problems, have led to the offer of a combined medical and psychological intervention oriented towards a global and integrated approach to the needs of a family that is changing with the arrival of a new member. Therefore, a clinical evaluation of the health status of the child recently arrived in Italy, targeted at identifying the presence of medical or psychological conditions, is performed during the day spent at the day hospital. The authors present data and considerations emerging from their ongoing experience, which has already involved 113 children and 89 families. The majority of the children evaluated so far show acute physical conditions, requiring major attention and support from their parents. The authors believe that in order to organize an adequate intervention it is useful to consider the parents of these children as caregivers. The function of caregiver is identified at three levels: the care of the child as son/daughter (typical of the parental function), the care of the problems deriving from the specific preadoption experience that accompany the child in the new family, and the care of the sick child. The authors deem that the combined medical and psychological intervention adopted at present with these families constitutes a useful support to the caregiver function. PMID- 19288782 TI - [Caregiver Need Assessment: a questionnaire for caregiver demand]. AB - Many studies have focused on the importance of the informal care provided by caregivers and on its impact in terms of worsening quality of life and increased burden. The aim of the present study is to analyze the psychometric validity and reliability of the Caregiver Needs Assessment (CNA) questionnaire, which has been built to investigate the needs (related to assistance) perceived by caregivers of severely impaired patients, particularly in the first stages of their illness. METHODS: The CNA was administered to 226 family caregivers (24.3% males) of 197 patients (50.8% males) hospitalized for neuromotor rehabilitation after a stroke, head injury, Lateral Amyotrophic Sclerosis, Parkinson or other severely impairing diseases. RESULTS: The instrument was tested on a large sample (KMO = 0.83) of heterogeneous caregivers. Explorative and confirmatory factor analysis, performed on a two subgroup random subdivision of the sample, showed the presence of two factors with good internal consistency: the factor "needs of emotional and social support" (alpha = 0.765) and the factor "needs of information and communication" (alpha = 0.742). The structural equation modeling confirms the goodness of fit of the 2-factor structure (RMSEA = 0.073; SRMR = 0.1; CFI = 0.96). We observed a positive correlation (p < 0.01) between the factor "needs of emotional and social support" of the CNA and other questionnaires aimed at assessing psychological wellbeing, and between the factor "needs of information and communication" and the factor "needs for knowledge about the disease" of the Family Strain Questionnaire, showing good convergent validity. We also observed high Pearson correlation coefficients (0.942 and 0.965) between test-retest measurements of both factors in the CNA. CONCLUSION: The statistical analysis confirms the good psychometric properties of the CNA questionnaire. For its brevity and ease in compilation the CNA is promising practical tool aimed at assessing caregivers' needs in order to personalize a programme of psychological support, to measure it's outcome and to provide comparison of the different needs in different diseases. PMID- 19288783 TI - [Formal representation of the cognitive-behavioral assessment in obsessive compulsive disorder]. AB - Psychological assessment is a crucial and important part of psychologist's work. Despite this, tools utilized by clinicians are often inadequate and methods used to investigate and make decisions on the basis of collected data are even logically incorrect. In this paper two mathematical psychology theories (Knowledge Space Theory and Formal Concept Analysis) are employed to propose a method for collecting diagnostic information. The conjoint application of these two theories allows to build a hierarchical structure composed by diagnostic elements. This structure is very similar to the one determined by the clinical diagnostic process. Correct logical inferences on the patient's situation, and an exact depiction of those diagnostic elements presented or not by the patient, represent the potential advantages of such a methodology. An applicative example on Obsessive-Compulsive Disorder (using the background determined by two shared diagnostic tools i.e. DSM IV-TR and CBA 2.0) is presented. Finally, the opportunity to build an adaptive assessment procedure from the derived structure is investigated. PMID- 19288784 TI - [Sleep disorders questionnaire for medical surveillance of workers]. AB - According to Italian law, occupational physicians should assess the fitness of employees for night work before their assignment, at regular intervals, and in cases of health issues related to night work. Moreover, sleep disorders among occupational drivers and shift workers need to be systematically investigated. Sleepiness at the wheel is now identified as one of the main reasons behind fatal crashes and highway accidents caused by occupational drivers. A significant percentage of workers suffer from sleep-disordered breathing, narcolepsy, sleep deprivation, poor sleep hygiene and circadian rhythm diseases. However, all these problems are underestimated. A questionnaire aimed at carefully assessing sleep disorders during medical surveillance of workers was carried out by the Italian Association of Sleep Medicine. It includes twenty-three questions and helps highlight any substantial sleep problem which could require further investigation by sleep medicine specialists. PMID- 19288785 TI - [The risk assessment procedures from SIMLII guidelines on upper extremity musculoskeletal disorders: discussion on results about a casistic from different working conditions]. AB - In this paper the result of application of SIMLII guide lines on upper extremity work related musuloskeletal disorders in different work conditions are presented and discussed. We applied the following risk assessment methods: Washington State Standard, HAL, OCRA, RULA, Strain Index, OREGE. The evaluations were subdivided in 3 class: acceptable risk, intermediate risk, high risk. It is confirmed that the application of more than one method for risk assessment, enable to better measure the risk and to focus the specific risk factors (frequency, posture, force, complementary factors). PMID- 19288786 TI - [Evidence based prevention and upper limb work-related musculoskeletal disorders]. AB - To evaluate interventions for primary prevention of Upper limb Work-related Musculoskeletal Disorders (UWMSD) we conducted a literature search from the biomedical database Medline and the Cochrane Collaboration Occupational Health Field. A total of 41 studies were selected: the majority investigated the effect of interventions among office workers, few involved industrial workplaces. Studies were characterized by a wide range of interventions (engineering, administrative, ergonomic training) and methodological heterogeneity (in the study design and outcome measures). Only four studies examine interventions for the prevention of specific outcomes (Carpal Tunnel Syndrome and Hand Arm Vibration Syndrome). At present, the multidimensional approach of interventions and the poor outcome definitions hamper the isolation of the potentially effective component of the intervention. Future intervention studies should be based on well defined risk assessment and outcome measures, rigorous and long term study design. Only strong levels of evidence could be the base of policy recommendations. PMID- 19288787 TI - [Working tasks with upper limbs repetitive movements: analysis of different methods for risk assessment]. AB - A review of different methods for the risk assessment of upper limbs repetitive movements is carried out mainly referring to a recent ISO standard (ISO 11228-3). This standard establishes ergonomic recommendations for tasks involving manual handling of low loads at high frequency (repetitive work). It is a "voluntary" standard and provides information for all professionals involved in occupational prevention as well as in job and product design. It refers to a four-step approach, involving both risk assessment and risk reduction (hazard identification, risk estimation, risk evaluation and risk reduction). General reference is made to a general model reported in a Consensus Document published by the IEA Technical Committee "Musculoskeletal Disorders", with the endorsement of ICOH. Apart from risk identification, the standard addresses and suggests several methods for a simple risk estimation (i.e. Plibel, Osha Checklist, Upper Limb Expert Tool, Qec, Checklist Ocra). If the risk estimation results in the 'yellow' or 'red' zone, or if the job is composed by two or more repetitive tasks, a more detailed risk assessment is recommended. For a detailed risk assessment, the OCRA method is suggested as "preferred"; however, other methods (STRAIN INDEX; HAL-TLV-ACGIH) can also be used. The applicative limits of the methods mentioned above, considering the purposes of the standard, are shortly discussed together with their recent scientific updates and applicative perspectives. The standard, with the suggested risk assessment procedures and methods, represents a useful tool for all OSH operators involved in the application of European and National legislation regarding the prevention of UL WMSDs. PMID- 19288788 TI - [Neck and upper limb disorders caused by combined exposures to ergonomic risk factors and hand-transmitted vibration]. AB - A review of neck and upper limb disorders caused by combined exposures to hand transmitted vibration and ergonomic risk factors (repetitiveness, force, posture) suggested the following conclusions: (1) hand-transmitted vibration has a dominant role in the etiopathogenesis of Raynaud's phenomenon and various forms of peripheral neuropathy with sensory impairment (digital, multifocal) in users of vibrating tools; (2) vibration of low frequency and high amplitude from percussive tools concur, together with adverse ergonomic factors, to produce degenerative changes in the bones and joints of the upper limbs, mainly in the wrist and elbow; (3) there is strong epidemiological and experimental evidence that combined exposures to hand-transmitted vibration and physical load are associated with an excess risk of carpal tunned syndrome; (4) there is limited evidence for an association between Dupuytren's contracture and vibration exposure owing to the small number of currently available epidemiological studies; (5) there is insufficient evidence for a contribution of hand transmitted vibration to the development of chronic pain and clinical syndromes in the neck and upper limb, while excessive physical load and ergonomic stress have a primary role in the etiopathogenesis of these disorders. PMID- 19288789 TI - [Musculo-skeletal disorders of the upper limb in the "new tables of professional diseases in industry and agriculture"]. AB - The author, after an analysis of the statistical data on work-related diseases reported to the INAIL and listed on the Annual Report 2007, examines the main highlights of the "new tables of professional diseases in industry and agriculture" published in the Ministerial Decree of July 21st, 2008 (GU n.169, 21 7-2008), also relating to the introduction of musculo-skeletal disorders of the upper limb due to bio-mechanical strain into the list of professional diseases to which the legal presumption of origin is applicable. PMID- 19288790 TI - [Smoking ban observance in a general hospital]. AB - Passive smoking is one of the main risk factors for severe chronic diseases. Hospitals are appropriate settings for promoting tobacco smoking cessation and preventing passive smoking. Moreover, since 2003 Italian Law (no 3/2003) forbids smoking in enclosed workplaces. The smoking ban should be strictly observed in hospitals. This study assessed smoking ban observance and exposure to ETS in a group of paramedical personnel working in a General Hospital (n=1037). Each subject answered a self-administered questionnaire. The prevalence of smokers was 31.2% in females and 31.5% in males. 83% of non-smokers and 66.6% of smokers approved the smoking ban in the hospital but 39.6% of non-smokers and 17.4% of smokers stated it was not observed. Percentages of, respectively, 57.0% and 31.1% in 2004 had dropped in 2005 and in 2006, after Law no 3/2003 was implemented. More than 90% of paramedical personnel declared that workers usually smoked in the workplace. 11.1% of non-smokers reported they were often exposed to ETS and 50% sometimes. These result show the smoking ban was not fully observed in the Hospital, where many workers reported they were exposed to ETS. Occupational Physicians should encourage health workers to stop smoking by offering appropriate counselling. PMID- 19288791 TI - [Acute and chronic alcohol abuse and work]. AB - A not moderate alcohol consumption or its abuse have relevant consequences not only on the health of the general population but also on the possibility to carry out any work in safety conditions. These behaviours have focused the attention of the institutions, which have promoted in the last years a growing number of preventive and informative actions and have adopted specific laws that have significantly involved the figure of occupational physician. Over the clinical implications, in fact, those behaviours, in the employment context, are associated with an increased risk of injuries (from 10 to 30% of total), an increase in the number of absences from work, with greater precariousness, with the possible interaction and/or strengthening of other occupational toxics and with the progressive reduction of working capacity. Diagnostic tools available for the detection of alcohol abuse or dependency consist, in acute cases by direct measuring of alcohol on blood, saliva and exhaled air, while in the chronic situations in addiction to the more traditional indicators (AST, ALT, GGT, MCV) there are recently introduced marker (CDT)--or in validation (ethyl glucuronide)--that representing, also with specific questionnaires (AUDIT, MAST, MALT, CAGE), useful integrated tools in the clinical-diagnostic path. The role and contribution of occupational medicine in the management of alcohol related problems is vital and relevant. Must be clear however that these are problems associated with a particular behaviour of the person and not with risks present on work-site. PMID- 19288792 TI - [Evaluation and management aspects of drug-abusers in the workplace]. AB - In Italy a recent legislation introduce detailed procedures to detect drug abusers, which had to be carried out in work settings. The purpose of the legislative document is to reduce the risk of work-related accidents which may affect both the working and the general population. In fact for the time being the procedure is foreseen for only a few work categories with a higher level of responsibility for third parties. This action is based on the adoption of the indications of the EU Drugs Action Plan (2005-2008), and it is mainly oriented to prevention through an early detection of occasional drug abusers to hopefully reduce the number of habitual drug consumers. Firm proprietors and manager are responsible of the overall procedure and the occupational physicians are responsible of the application of many parts of it. The paper discuss many aspects related to the application, including: physician responsibility against the law, how to carry out efficient and accurate drug testing, collaboration with other public agencies devoted to rehabilitation of drug-abuse patients, methodological aspects that can allow the procedure to be effective. PMID- 19288793 TI - [Determination of absence of drug dependence: procedures to follow and role of the surveillance body]. AB - The assessment of presence or absence of substance addiction in subjects working in activities potentially harmful for themselves or others, is indeed an important issue and a compulsory activity. Nevertheless, according to Italian laws (outlined in the agreement on the ascertainment of the absence of drug addiction, executed in the session of October 30th 2007), only occupational company doctors are in charge of such a duty. This professional figure should instead be mainly aimed to protect worker's health. Furthermore, private workers, even in settings or jobs at risk (e.g. bus drivers) are automatically excluded from such a verification. In this case, third parties (e.g. bus passengers) are protected by the law only when the worker is employed in a company. The new procedures, recently promulgated by central and regional governments, increase the burden of occupational company doctors, and introduce restrictions for workers of particular activities, in the case of a positive laboratory result for prohibited substances. PMID- 19288794 TI - [Level II diagnosis and rehabilitation programs: role of drug dependence services]. PMID- 19288795 TI - [The European regulation REACH: role of the toxicologist and the occupational specialist]. PMID- 19288796 TI - [Occupational health and safety management systems: scenarios and perspectives for occupational physicians]. AB - This paper analyzes the role of the occupational physicians, taking into account the new Italian legislation within the frame of CSR, that puts in a new light the physicians inside the Organizations. In this context, Occupational Medicine and Workplace Health Promotion play a central role in most of the items of the Occupational Health and safety management systems, from H&S politics to training, from First Aid to audit and revision systems. From this innovative perspective, the authors try to identify the occupational physician's new challenges and opportunities. PMID- 19288797 TI - [Occupational physician's role in the prevention of the accidents in construction industry]. AB - The aim of this work is to discuss about the role of the occupational physician in the prevention of the accidents in construction industry. Using the experience of 12 years of surveillance of workers in Bergamo province, the authors analyse the "human factors" which may influence the risk to have an accident, and the role of the physicians not only for the early diagnosis of work-related diseases, but also for the formulation of correct fitness to work, which consider accidents' prevention. Health conditions, psychological elements, fatigue and life style are some of the most important "human factors" which can amplify the accident phenomenon in construction industry. Our experience demonstrates that the occupational physicians can operate in preventive way on these factors, formulating correct fitness to work, giving their collaboration in the risk evaluation and management, suggesting runs of rehabilitation and recovery for the workers who need it, promoting information meetings related to the correct life habits. PMID- 19288798 TI - [Methodology of risk assessment by the occupational physician at the light of new legislative rules]. AB - The occupational physician may play a crucial role in the risk assessment of workers in a given occupational environment. In fact, according to the current italian legislation, the document on risk assessment must be produced by the employer, with the cooperation of the responsible for prevention and safety and, in the due contexts, with the contribution of the occupational physician. Therefore, a cooperation between the different figures involved in the process is a pre-requisite for the elaboration of an adequate planning of preventive measures. From the point of view of the occupational physician, it seems necessary a preliminary in-depth knowledge of the plant and of the various steps of the productive process. In particular it is mandatory to have precise data on the number of workers, the mean age, work seniority, and the percentage of disabled workers. As a corollary, it is important to know the frequency of occupational related disorders, the number of accident at work, and the possible presence of contentious procedures with security organisms. Several important open questions remain, however: in particular to establish a causal relationship of any disorder with occupational situtations and to discriminate between distressing situations inside and outside the work environment. PMID- 19288799 TI - [Changes in work organization and management of psychosocial risk factors]. AB - In recent years, major changes have occurred in Italian working conditions and employment patterns due to several concurrent factors: increasing occupation in the tertiary sector, implementation of new technologies, labour market globalization, higher variability of working time arrangements, decrease of traditional physical-chemical risks, ageing of general/working population, access to work of people with disabilities, growing immigration of extra-community workers. Thus, psychosocial risk factors are becoming crucial issues of the present work organization, dealing with job content (complexity, meaning, uncertainties), mental work load, time pressure, variable working hours; career perspectives, role conflicts and ambiguity, education and training, personal relations, social support, work/family conflicts; age and cultural discrimination. The Occupational Health Physician has to deal with these multidimensional and multifaceted aspects of work stress by different and concurrent approaches, at both group and individual levels, with epidemiological and clinical perspectives, enacting preventive and therapeutic strategies. Both "external" work load and individual "responses" have to be properly considered and risk has to be assessed with "relative" rather than "absolute" criteria, addressed not only at fitness to work, but also to corrective actions. Hence, the OHP has to act in closer collaboration with work psychologists, sociologists, human resources managers and work organisation experts. PMID- 19288800 TI - [Contribution of research to the responsible and sustainable development of nanotechnologies]. AB - The newly-fledged nanotechnologies offer opportunities for social development but uncertainties prevail about their impact on human and environmental health. Right now there is still a huge gap between technological progress and research into the health and safety aspects of nanomaterials. This is clear from the quantity of nano-products already on the market--more than 600--and the public and private funds dedicated to the development of nanotechnologies, which are almost a hundred times those available for research into their effects on health and safety. Estimates have it that by 2014 nanotechnologies will be widely used in our society, and ten million new jobs will be created. Therefore it becomes essential to plan an integrated approach to specific risk analysis at work. The following gaps and needs come to light: limited information; difficulties in relating nanotechnologies and production of nanomaterials to specific areas of application; efforts required to assess the hazards posed by nanomaterials in realistic exposure conditions; ethical issues about nanotechnology in the workplace likely to arise from today's knowledge about the hazards of nanomaterials and the risks they may pose to workers. An integrated approach to research, cooperation and communication strategies is essential if we are to direct our efforts towards responsible and sustainable growth of nanotechnologies. PMID- 19288801 TI - [Molecular diagnosis of lung cancer]. AB - Individuals who have been exposed to lung carcinogenics are concerned by their greater risk of developing lung cancer, thus consult physicians with the expectation of undergoing screening tests. Controversy remains as to which screening procedures should be assessed. Previous studies of sputum cytology and chest X-rays showed no benefits in lung cancer mortality reduction. Screening of high risk individuals through computerized tomography scan appeared promising, but this radiological technique suffers from low specificity. Currently, the differential diagnosis is mainly based on additional imaging techniques, particularly positron emission tomography, which is expensive, and also shows limitations in terms of sensitivity and specificity. Therefore, purely morphological criteria seem to be insufficient to distinguish lung cancer at early stages from benign nodules with sufficient confidence, and false positives undergoing surgical resection seem to occur frequently. A molecular approach to the diagnosis of lung cancer through the analysis of biomarkers obtained by non invasive means could greatly improve the specificity of imaging procedures. Extremely sensitive mass spectrometric techniques are available to detect molecular alterations in accessible media, which characterise lung cancer at early stages, thereby reducing the rate of false positives, expected to be very out without a sound application of confirmatory diagnostic tests. PMID- 19288802 TI - [Coordination patterns assessed by a continuous measure of joints coupling during upper limb repetitive movements]. AB - We analyzed the coordination patterns between elbow, shoulder and trunk in a motor task consisting of reaching out, picking up a cylinder, and transporting it back by using the Dynamical Systems Theory and calculating the continuous relative phase (CRP), a continuous measure of the coupling between two interacting joints. We used an optoelectronic motion analysis system consisting of eight infra-red ray cameras to detect the movements of nine skin-mounted markers. We calculated the root square of the adjusted coefficient of determination, the coefficient of multiple correlation (CMC), in order to investigate the repeatability of the joints coordination. The data confirm that the CNS establishes both synergic (i.e. coupling between shoulder and trunk on the frontal plane) and hierarchical (i.e. coupling between elbow-shoulder-trunk on the horizontal plane) relationships among the available degrees of freedom to overcome the complexity due to motor redundancy. The present study describes a method to investigate the organization of the kinematic degrees of freedom during upper limb multi-joint motor tasks that can be useful to assess upper limb repetitive movements. PMID- 19288803 TI - [Prospects of development for occupational surveillance and epidemiology]. PMID- 19288804 TI - [Radioprotection: doctrinaire references and medical surveillance]. AB - The current statement of radioprotection, as formulated in the 60/90 ICRP Publication and confirmed in the recent 103/07 ICRP Publication, strengths the conceptual principle of the optimization, justification and limitation of doses, it analyses the risk and proposes the values of dose limitation, according to socio-medical considerations. The medical radioprotection's aim moved from the limitation of deterministic damages ("tissue reactions" according to ICRP 103/07) to the probability's reduction of stochastic effects appearance. The "tissue reaction problem" was solved maintaining the exposition limits under the threshold. All the Radiological Protection System is orientated toward the reduction of stochastic effects appearence based on epidemiological evidences (if findable) and caution: caution is represented by LNT (Linear No-Threshold Hypotesis). This hypothesis permits to apply criteria of risk's evaluation by mathematics intruments. Even if sometimes this hypothesis was scientifically criticized recently (ICRP 103/07) it's been considered valid for the prevention in the radiological protection sphere. The medical radioprotection is interested in worker's general state of health. This state of health should be compatible with specific work's conditions and permit to formulated a judgement of fitness. This compatibility (without contraindication) shall be kept and checked time by bringing about preventive measures and, in the same time, picking out the first signs of any pathologies. Therefore with the radioprotection we can talk about genetic individual susceptibility to neoplasia. Especially with the moleculer genetics we can recognize a quite big number of hereditary defects combined with family predisposition to cancer. The medical surveillance therefore has to be a prevention activity for each individual worker considering his ipersusceptibility, his specific working risk, his particular state of health, his habits, his family predisposition, his aspirations and his socio-cultural context. PMID- 19288805 TI - [The health protection policy]. PMID- 19288806 TI - [Radio epidemiological data and genetic susceptibility to radiation-induced cancer given in the new ICRP recommendations]. PMID- 19288807 TI - [Radon and internal contamination]. AB - Because of hits everywhere presence in air and in water needful mediums for life, radon is a omnipresent risk for every person. Therefore, in relation to those vital functions, lungs and gastro-enteric tract represent the principal target organs of this noble radioactive gas (and mainly of hits radioactive daughters). International organisms evaluated the effective dose coefficients for both target organs, so it is possible e quantitative assessment of the exposure risk related to this noble gas. PMID- 19288808 TI - [Diagnostic protocol and special tests]. AB - Diagnostic protocol and special tests to ionizing radiations have a preventive function in the medical surveillance of the exposed worker. This protocol must be provided with laboratory and special test assessing fitness for working at the risk of ionizing radiations. The health of workers must be compatible with working conditions and radiation risk. This healthiness of workers is evalued in the time to give an assessment fitness for working at ionizing radiations. For this purpose the basic diagnostic protocol must guarantee minimum information about state of organs and apparatus in addition to the normality of the metabolisms. The diagnostic protocol of the exposed worker to ionizing radiations must be adapted to the specific clinical situation so as to finally make a costs benefits balance sheet. PMID- 19288809 TI - [Judgement of fitness to work and forensic-medical aspects]. AB - The occupational physician, specifically trained in radiation protection, has to consider different criteria, such as psycho-physical ability, biological fitness and worker's validity to formulate judgement of fitness to work in exposure to ionizing radiation (IR). According to National Legislations (D.Lgs. 230/95; DM 488/2001), there are different possible outcomes about fitness to work criteria: absolutely fit to work, partially fit to work, not fit to work and medical examination before workers exposed to IR's retirement; besides, the occupational physician has to pay particular attention to evaluate individual hypersusceptibility conditions. Stochastic somatic diseases require a "not fit to work" judgement, an occupational disease complaint to the Insurance Institute and a report obligation. Forensic-medical ascertainment of diseases caused by radiation exposure needs different criteria, particularly for stochastic effects. The most suitable criterion, applied to attribute etiologic role in stochastic effects, is "Probability of Causation" theory, deriving from scientific literature evidence. This theory is an epidemiology-based method conceived to evaluate the probability of a linkage between a specified cancer and an exposure. Judgement of fitness to work in case of neoplasia partial or complete remission should be based on objective considerations, giving particular importance to the human and social aspects. PMID- 19288810 TI - [Medical and risk records]. AB - Aim of this report is to outline the importance of personal medical records collected from the occupational health surveillance practice. The Italian law D.Lgs. 81/08 puts a great emphasis on a national information system built up by health monitoring results in workplace as far as individual and general prevention is concerned. Private and public roles and duties are discussed. PMID- 19288811 TI - [Register of exposed to carcinogens]. AB - Law Decree 626/94 already provided for the "register of exposed subjects" to be implemented by employer once the presence of cancerogenic agents falling into the definition of the norm itself have been assessed in working activities. The lack of an effective decree identifying the models and modalities of register keeping has led so far to an exposure under-evaluation. Of course the coming into force of Ministerial Decree 155/2007, almost at the same time as the new body of regulations made up by Law Decree 81/2008, stimulated the addressees to proceed with setting an exposure register to be forwarded to the National Institute for Occupational Safety and Prevention (ISPESL) and to the competent inspection authority. Starting for the regulations, the authors will go through the register forms and information flow management. PMID- 19288812 TI - [Periodic health report by occupational physician: tools and contents for the writing]. PMID- 19288813 TI - [The occupational and active research, diagnosis and reporting of occupational diseases]. AB - The Occupational Doctor plays a key role on the protection of workers safety and prevention of occupational and work-related disease. His training and his need of updating, due to the constant transformation of work, the evolution of technology and the medical progress, set him in the center of all activities carried out to protect health in workplaces. The Occupational Doctor should devote special attention to the working anamnesis as well as to the pathological anamnesis both close and remote. He should also pay attention to semeiotics. Another important element is the relationship between the Occupational Doctor and the Doctor of General Medicine (MMG); this synergy is of vital importance in protecting health and in investigating diseases whether occupational or not. D.Lgs. 81/08 emphasizes this synergy, in fact name and phone number of Doctor of General Medicine is compulsory in Case History. Major source of information for all form of prevention is the survey of occupational disease which is a tool for epidemiological control. The use of a systematic collection of data, of protocols, of guidelines and of scientific evidence is the basis for identifying occupational diseases, their diagnosis and subsequent denunciations. This is the line suggested in MAL PROF informative system, made for registration of work related diseases, and which is important, with other instruments, in realizing an integrated informative system for prevention in workplace. The Covenant for the Protection of Health and Prevention in Workplaces (DPCM 17/12/2007) indicates the strategic objectives of the National Health System for the consolidation and development of the existing system and of the programs promoting health and safety. These include the growth of the culture of prevention and of the epidemiological control of occupational diseases. The Occupational Doctor has an important role because he can identify the early onset of diseases during the working age and start all forms of prevention and health promotion. In the case of diagnosis of a suspected professional disease the Occupational Doctor has three distinct obligations. The first is to report to the legal authority (C.P.P. art. 365). The second requirement is the declaration of the occupational disease (D.P.R. 1124/65 art. 139) and the third is to issue the first certificate of occupational disease for compensation insurance (D.P.R. 1124/65 artt. 53, 251). PMID- 19288814 TI - [The role of "competent physician" for prevention of accidents at work]. AB - Using at best the professional and legal tools at his/her disposal, the "competent physician" can have a relevant role in reducing accidents in the workplace. In assessing the worker's suitability to specific tasks, the competent physician checks the presence of pathologies or functional impairments of organs or apparatuses that can be an additional risk for the occurrence of accidents at work. The activity aimed to preventing accidents at work translates also in taking part in the planning and implementation of information activities and workers' training within the company. The various risk factors inherent in the workplace and so-called "human factors" can interact in a negative way to the point of becoming a cause of accidents. The human variables of accident risk at the industrial, occupational and even individual level, may be numerous. In this paper we shall review these important aspects and attempt to clarify the role that can be played by the competent physician in the prevention of accidents at work. PMID- 19288815 TI - Surgery of the mediastinum: historical notes. AB - One century ago, thoracic surgery was in its infancy. Since then, advances in chest radiology have allowed for reliable and early detection of thoracic ailments. Refinements in anesthesiology and surgical technique have led to the development of a plethora of techniques for the diagnosis, staging, and treatment of mediastinal disease. As the disease processes we encounter have expanded and evolved, so too has our surgical armamentarium. PMID- 19288816 TI - Utility of positron emission tomography in the mediastinum: moving beyond lung and esophageal cancer staging. AB - PET and PET-CT are rapidly evolving as modalities of thoracic imaging. In the mediastinum, PET can provide information to distinguish thymic hyperplasia from neoplasia, although the use of this imaging for this purpose is not accepted uniformly as necessary. PET is the standard of care in staging and follow-up of mediastinal lymphoma and in follow-up of metastatic seminomas after chemotherapy. Mycobacterial/fungal infections, sarcoidosis, and brown fat can mimic malignant findings on PET in the mediastinum. PMID- 19288817 TI - Genetic markers of mediastinal tumors. AB - Determination of the genetic markers by the application of new genomic methodologies has provided important insight into the pathogenesis of mediastinal disease. These new techniques have enabled scientists to uncover differential gene expression patterns between subtypes of thymomas, correlate tumor marker expression with germ cell tumors, and determine a link between the NF-kappaB and JAK/STAT pathways with Hodgkin's and non-Hodgkin's lymphoma. Despite the progress made in the understanding of genetic markers of select mediastinal tumors, significantly more investigation is required to elucidate the molecular pathways involved in the pathogenesis of these tumors. PMID- 19288818 TI - Diagnostic strategies for mediastinal tumors and cysts. AB - Many histologically different tumors and cysts that affect people of all ages arise from the multiple anatomic structures present in the mediastinum. The number of diagnostic possibilities can be narrowed by considering the patient's age, tumor location, the presence or absence of symptoms and signs, the association of a specific systemic disease, radiographic findings, and biochemical markers. Pathologic diagnosis is often required to confirm a presumed diagnosis and to select the optimal treatment modality. A variety of biopsy techniques for obtaining tissue from the mediastinum have been described, including ultrasound-guided endoscopic biopsy, percutaneous image-guided needle biopsy, parasternal anterior mediastinotomy, cervical mediastinoscopy, and video assisted thoracoscopic surgery. The choice of biopsy technique depends on the localization of the lesion, clinical factors such as the age and the condition of the patient, and the availability of special techniques with the required expert and the equipment. PMID- 19288819 TI - Infections of the mediastinum. AB - Infections of the mediastinum (ie, mediastinitis) are serious, are associated with high morbidity and mortality, and may result from adjacent disease with direct extension, hematogenous spread, or direct introduction into the mediastinal space. The organs and tissues involved determine the manifestations and approach to treatment of these infections. The most common ones are those secondary to perforation of the esophagus or penetrating trauma, and those that extend from an adjacent infection. Today, the most common cause of mediastinitis is direct invasion of the mediastinum after surgical intervention. Cases of mediastinitis can be classified as either acute or chronic. Two broad categories of acute mediastinitis are acute necrotizing mediastinitis and poststernotomy mediastinitis. Chronic mediastinitis has been arbitrarily subdivided into two categories: (1) granulomatous mediastinitis, and (2) fibrosing or sclerosing mediastinitis. However, these likely represent a continuum of chronic infection. In cases of acute mediastinitis, treatment should always be directed toward the primary pathology and the clinical presentation. In chronic cases, surgical treatment is only palliative. PMID- 19288820 TI - Mediastinal tumors and cysts in the pediatric population. AB - Pediatric mediastinal tumors and cysts are rare disorders that share many similarities with adults, yet which have important differences unique to the child. Posterior mediastinal tumors are relatively more common in children than in adults and are also more likely to be malignant in children. CT imaging facilitates the diagnostic evaluation of mediastinal masses in children. Airway compression is always a concern with large mediastinal tumors in children given their relative softer and smaller airway. PMID- 19288821 TI - Multimodality treatment of germ cell tumors of the mediastinum. AB - Germ cell tumors originating in the anterior mediastinal compartment represent a rare but biologically interesting group of neoplasms. Knowledge of the specific biologic behaviors and therapeutic strategies for the three histologic types is important. PMNSGCT represent the most challenging group of malignant germ cell tumors and survival outcome is dependant on both successful chemotherapy and surgery to remove residual disease when feasible. The authors currently believe nonbleomycin-containing regimens will reduce operative risks in this regard. New chemotherapy strategies that reduce the incidence of persistent nonseminatous germ cell or non-germ cell cancer need continued investigation. Although overall survival is inferior to nonseminomatous germ cell tumors of testicular origin, favorable subsets with pathologic evidence of either necrosis or teratoma have been identified. An aggressive surgical approach after cisplatin-based chemotherapy can result in long-term survival, even in patients with persistent nonseminomatous germ cell or non-germ cell cancer, and is warranted in these otherwise young and healthy patients. PMID- 19288822 TI - Multimodality treatment of thymic tumors. AB - Combined modality therapy is gaining acceptance for treating stage 3 and 4A thymic tumors. Also, specific subsets of stage 2 tumors deserve particular attention. Single-center experiences demonstrate that there are some advantages in selected groups of patients. The overall relatively low complete response rate, however, imposes the search for better systemic therapy to optimize results. In fact, although thymic tumors are responsive to different cytotoxic regimens, none has been demonstrated to be the ideal one. New therapies and strategies should be designed and tested in large-scale multicenter prospective trials. Among the others, epidermal growth factor receptor inhibitors have shown some clinical response, because EGFR is overexpressed in thymoma. c-KIT is overexpressed in thymic carcinoma. Although in a recent study a clinical response to imatinib has been reported, results of a prospective study in patients who have thymic carcinoma are pending. Clinical responses have been reported also to other tyrosine kinase inhibitors, such as dasatinib. Other reports have stressed the presence of an up-regulation of COX-2 with a potential separate therapeutic pathway. Other markers, such as the expression of thymidine synthase and dihydropyrimidine dehydrogenase, which predict sensitivity to 5-fluoruracil-based chemotherapy, were not correlated with the clinicopathological characteristics in a series of thymomas. These new therapies should be incorporated in a standardized approach that goes from a careful assessment of histology, staging, and lymph node status, and a constructive and nonempiric cooperation between the oncologist, radiotherapist, pathologist, and thoracic surgeon. PMID- 19288823 TI - Surgical approaches to the thymus in patients with myasthenia gravis. AB - Myasthenia gravis is an autoimmune disorder of neuromuscular transmission affecting 2 out of every 100,000 people. Neurologists and surgeons still debate what role surgery should play in its management. Many patients who might benefit from thymectomy are denied the opportunity because of misconceptions, ignorance, or trepidation. By offering effective methods of less invasive thymectomy to these patients, a significant number of patients and treating neurologists previously unwilling to consider surgery may realize the benefits of this established, proven treatment alternative. The surgical approaches reviewed include: transcervical, videothoracoscopic, robotic-assisted, transsternal, and combined transcervical-transsternal maximal thymectomy. PMID- 19288824 TI - Vascular lesions of the mediastinum. AB - Most mediastinal masses seen on thoracic imaging are of solid tumor origin. As such, vascular lesions of the mediastinum can be mistaken for, or misinterpreted as, one of these more common mediastinal masses. This article stresses the importance of considering vascular entities in the differential diagnosis of mediastinal masses. PMID- 19288825 TI - Combined cervicothoracic approaches for complex mediastinal masses. AB - Tumors or masses occurring in the cervicothoracic region show a broad variety in their histologic origin. Accordingly, the indication for surgical resection varies, and especially in malignant lesions, surgery frequently is part of a multimodality treatment. Adequate access is important for any operation in a critical region such as the cervicothoracic junction. The anatomic location of the mass to be resected usually dictates either an anterior or posterior approach. The need for appropriate exposure is in addition paralleled by the need to preserve a maximum of important functional structures, to maintain their diligent functional interaction. Careful interdisciplinary preoperative treatment planning is necessary to achieve optimal results. PMID- 19288826 TI - Intraoperative strategy in patients with extended involvement of mediastinal structures. AB - The mediastinum is a virtual space containing several vital organs and structures. Biopsy and resection of lesions located within this region often require several considerations that bear on intraoperative strategy. To optimize outcome, clinicians must be able to predict which patients are at highest risk of anesthetic complications. Superior vena cava involvement, extensive compression of the airway, and pericardial effusion have a clear impact on the decision making of the anesthetist and surgeon, who should plan together when forming the surgical strategy. PMID- 19288827 TI - The role of surgery in recurrent thymic tumors. AB - Surgery plays an important role in treating recurrent thymic tumors. This approach may require both induction therapy and oncological consolidation (adjuvant) in case the lesion shows extended invasion of the surrounding organs. Extended resections are justified to achieve complete removal of the recurrence. Recurring thymic carcinoma or carcinoids may be more difficult to treat. PMID- 19288829 TI - Business 101. PMID- 19288828 TI - Advances in radiotherapy for tumors involving the mediastinum. AB - RT is an integral component of treatment in many tumors of the mediastinum. In recent years, there have been advances in RT planning and delivery, which have allowed more effective radiation delivery while sparing normal tissues. New technologies and radiation modalities may help achieve disease control more safely and effectively. PMID- 19288830 TI - Dental morality or misguided science? PMID- 19288831 TI - Smile design and advanced provisional fabrication. AB - Cosmetic reconstruction is much more predictable when superb provisionals are used as a blueprint for the final restoration. This article provides details for a technique that can be used to fabricate indirect multilayered composite provisional restorations for porcelain veneers and complex full-arch cases. PMID- 19288832 TI - Association of enamel microabrasion and bleaching: a case report. AB - Enamel microabrasion may be indicated for certain clinical situations, since it is a proven method for removing superficial intrinsic enamel discoloration defects such as fluorosis-like stains; in addition, it is safe, conservative, and easy to perform. However, this method removes enamel structure, causing teeth to become yellowish. This yellowing can be treated with enamel microabrasion and bleaching. This article describes and documents how these two methods were utilized to correct tooth color for a young patient. PMID- 19288833 TI - Facial appearance following surgical treatment for obstructive sleep apnea syndrome. AB - Obstructive sleep apnea syndrome (OSAS) is a serious medical condition that is associated with numerous negative health side effects. The general dentist plays an invaluable role in identifying patients with this condition. Certain OSAS patients receive significant medical and social benefits from orthognathic surgery to advance the maxilla, mandible, and chin. Anterior positioning of the maxilla and mandible is not only highly successful for curing OSAS but also results in favorable facial esthetic changes. PMID- 19288834 TI - Esthetic restorative material shade changes due to photopolymerization. AB - This study sought to measure color values (before and after photopolymerization) and variations of A1 shade polymeric dental restorative materials. The L*a*b* values of polymeric dental restorative material disks were measured by a spectrophotometer with the specular component excluded (SCE) geometry under D65 illumination over a standardized white background before and after photopolymerization. Color differences (DeltaE*ab) due to polymerization were calculated using the CIE Lab color-difference formula. DeltaE*ab indicates differences between two colors in the L*a*b* color space. In this system, L*, a*, and b* indicate lightness, red-green, and yellow-blue, respectively. Color differences (DeltaE*ab) were calculated by the equation DeltaE*ab = [(DeltaL*)2 + (Deltaa*)2 + (Deltab*)2]1/2. Eight resin composites, three flowable resin composites (FRCs), and two resin-modified glass ionomers (RMGIs) were utilized; L*a*b* and DeltaE*ab values of the test groups were analyzed using ANOVA and Newman-Keuls multiple comparison tests. DeltaE*ab values before and after photopolymerization ranged from 3.01-17.76. All RMGI and two FRC materials displayed greater color changes than the resin composites (p < 0.05). Photopolymerization produced measurable increases and decreases in lightness. Yellow-blue (b*) values decreased in all test groups after polymerization, while most red-green (a*) values increased. Color differences between polymerized A1 shade materials ranged from 0.76-25.41 DeltaE*ab. The color difference between the test materials and a widely used tooth shade guide (the Vita Lumin) was averaged at 12.66 DeltaE*ab. PMID- 19288835 TI - Optimizing esthetics and function through interdisciplinary dentistry. AB - Esthetic dentistry has evolved dramatically over the years to the point of becoming an expectation from most patients. It is not a fad but is becoming a standard of care for the majority of dental treatment. By learning to practice good quality interdisciplinary dentistry, it is possible to turn a general practice into a comprehensive care practice that caters to high-quality esthetics and function. This article discusses incorporating interdisciplinary dentistry into the general practice and offers some clinical examples. PMID- 19288836 TI - Effects of 10% carbamide peroxide treatment and sodium fluoride therapies on human enamel surface microhardness. AB - This study sought to evaluate the microhardness of enamel submitted to 10% carbamide peroxide treatment and different methods of remineralization involving sodium fluoride (NaF). Non-erupted third molars were used and 75 enamel blocks with standardized dimensions (4.0 x 4.0 x 2.0 mm) were obtained. Enamel blocks were randomly divided into five groups (n = 15): one control (no bleaching and no fluoride treatment), one receiving a 10% carbamide peroxide treatment, one receiving 10% carbamide peroxide plus acidulated phosphate fluoride, one receiving 10% carbamide peroxide with 0.2% NaF, and one receiving 10% carbamide peroxide and 0.05% NaF. There were no differences among four of the groups (p > 0.05); the group receiving 10% carbamide peroxide only was different from all other groups (p < 0.05). Based on these results, NaF therapies are recommended during carbamide peroxide bleaching treatments. PMID- 19288837 TI - A technique for restoring Class V caries in primary canines. AB - This article describes a technique for restoring Class V carious lesions in the primary dentition. The discussion will include the treatment of aprismatic enamel in primary teeth, bonding to dentin, and the restorative techniques that are available for composite bonding. A case report is presented that utilizes several modern techniques for restoring Class V lesions. PMID- 19288838 TI - A multidisciplinary esthetic approach to single-tooth replacement and diastema closure. AB - Anterior tooth replacement has always posed an esthetic challenge for dentists. A multidisciplinary approach makes it possible to provide patients with esthetics and function. Treatment planning should involve other specialists when clinicians are faced with the additional esthetic concerns of the patient (such as anterior spatial anomalies and esthetically unpleasant existing restorations). Metal ceramic fixed partial dentures have been used to allow the replacement of missing teeth for many years, providing both strength and esthetics. Several other options also are available, including implant placement. Dental implants in the esthetic zone are well-documented in the literature; numerous controlled clinical trials have shown overall implant survival and success rates. This article illustrates a step-by-step analysis and treatment option for a single-tooth replacement and maxillary anterior spatial dilemma using Captek crowns. PMID- 19288839 TI - Treating tetracycline staining in the adult dentition: a case report. AB - Tetracycline staining in the adult dentition has presented an esthetic challenge for many years. Many clinicians have treated the problem by utilizing invasive techniques that may involve crowns, veneers, and composite bonding. Some major disadvantages to these invasive techniques include the cost involved and the need to cut on sound tooth structures. This article showcases a protocol for treating tetracycline staining non-invasively. PMID- 19288840 TI - Treatment planning in esthetic dentistry requires careful listening to the patient. AB - Creating an esthetic result that is pleasing to both the patient and the dentist requires careful consideration of patient concerns and treatment goals. This article presents a case involving a patient with many esthetic concerns. Following in-depth examination of the patient and consultations, an esthetic treatment plan involving the placement of laminate veneers was developed and executed. After the first set of laminate veneers was placed, some concerns led to an alteration in the proposed treatment plan. Each step of the treatment plan must be assessed before the next step is performed to guarantee a result that is both functional and esthetically pleasing. PMID- 19288841 TI - Clinical management of discolored teeth. AB - Discolored teeth are seen frequently in the dental office and present a major challenge to dentists. Discoloration may be limited to a single tooth or several teeth in a single arch or it may be generalized and evident on all of the teeth. It is essential to recognize the cause and to manage the discoloration accordingly. Generally, dentists' intervention is essential for treating substantial tooth discoloration. In some cases, scaling and polishing the teeth will improve the situation; however, more extensive treatment often is needed to achieve a satisfying result. Treatment options include vital and non-vital bleaching, microabrasion, composite and porcelain veneers, and porcelain crowns. Sometimes these treatments are combined for a more successful outcome. PMID- 19288842 TI - How to improve the health and well-being of the United States and the world by 2025 for only 3.1 trillion dollars a year. PMID- 19288843 TI - Using the RE-AIM framework to evaluate the statewide dissemination of a school based physical activity and nutrition curriculum: "Exercise Your Options". AB - PURPOSE: Examine the reach, efficacy, adoption, implementation, and maintenance of a physical activity and nutrition curriculum for middle-school students. DESIGN: Nonexperimental pilot evaluation of a statewide dissemination trial. SETTING: California middle schools during the 2006 to 2007 school year. SUBJECTS: Sixteen classes (N = 668 students and 16 teachers) sampled from the statewide pool who used the program. INTERVENTION: An eight-lesson nutrition and physical activity curriculum, "Exercise Your Options" (EYO), including a teacher guide, video clips, a student activity booklet, and ancillary materials was made available to teachers. MEASURES: Program records, classroom observations, teacher surveys, and student presurveys and postsurveys (assessing physical activity, sedentary behaviors, and dietary intake). ANALYSIS: Descriptive statistics and multilevel random-coefficient modeling. RESULTS: The EYO program reached 234,442 middle-school students in California. During the program, total physical activity increased (p < .001), whereas watching TV/DVDs and playing electronic games/computer use decreased (p < .05). Intake of dairy products increased (p < .05), whereas consumption of sugars/sweets decreased (p < .001). Forty-two percent of eligible middle-school classrooms ordered the program materials. Eighty-six percent of sampled teachers implemented all of the lessons. Over the past 5 years, 51% of all middle-school students in California were exposed to the program. CONCLUSIONS: The EYO program showed its potential for moderate to high public health impact among California middle-school students. PMID- 19288844 TI - Associations between general parenting styles and specific food-related parenting practices and children's food consumption. AB - PURPOSE: Explore the impact of general parenting style and specific food-related parenting practices on children's dietary habits. DESIGN: Cross-sectional study of sixth graders and their parents. SETTING: Data were gathered (in 2003) in 69 of 100 randomly selected elementary schools in Belgium. SUBJECTS: All sixth graders (N = 1957) were invited to participate; 82.4% of their parents gave consent and completed questionnaires, resulting in 1614 parent-child pairs. MEASURES: Children's consumption of breakfast, fruit, vegetables, soft drinks, and sweets was assessed by self-administered food frequency questionnaires. Parents completed questionnaires on sociodemographic characteristics, general parenting styles (authoritarian, authoritative, indulgent, or neglecting) and specific food-related parenting practices (pressure, reward, encouragement through negotiation, catering on children's demands, permissiveness, avoiding negative modeling, and praise). ANALYSIS: Logistic regression analyses were performed, with general parenting style and specific food-related parenting practices as predictors and dietary habits as dependent variables, controlling for sociodemographic characteristics and children's weight status. RESULTS: General parenting style did not show any significant impact on dietary habits. In contrast, the food-related parenting practice "encouragement through negotiation" showed a significant positive impact, whereas "pressure," "catering on demand," and "permissiveness" were practices with an unhealthy impact. CONCLUSION: Nutrition education programs that guide parents in firm but not coercive food parenting skills are likely to have a positive impact upon children's dietary habits. PMID- 19288845 TI - Factors which influence tobacco use among junior enlisted personnel in the United States Army and Air Force: a formative research study. AB - BACKGROUND: Despite the implementation of a host of tobacco control initiatives, tobacco use among active duty members of the U.S. Military remains high. It has been suggested that a positive culture of tobacco, which drives consumption, exists in the military. However, little is know about factors that impact tobacco use among military personnel. OBJECTIVE: This study provides the first formative data on the culture of tobacco in the U.S. Military. METHODS: A total of 15 focus groups on six military installations were conducted (n = 189). These military installations were located throughout the continental United States and were of average size for each service. RESULTS: Participants suggested that the primary method the military uses to discourage use is tobacco bans. Unfortunately, they also believed that the military accommodates tobacco use so that smoking remains convenient despite the bans on tobacco use. Smoking was believed to be encouraged through liberal smoking breaks, social interaction within designated smoking areas, and cheap and convenient tobacco products sold on military installations. Additionally, smoking was seen as an effective method to combat the stress and boredom of military life and to avoid weight gain. CONCLUSIONS: Suggestions for addressing the culture of tobacco are suggested. PMID- 19288846 TI - Contextual effects of built and social environments of urban neighborhoods on exercise: a multilevel study in Chicago. AB - PURPOSE: Examine the contextual effects of neighborhood built and social environments on exercise. DESIGN: Cross-sectional, multilevel study. SETTING: City of Chicago. SUBJECTS: A probability sample of Chicago adult residents (response rate = 55%). MEASURES: The exercise measures were based on two questions: "How often a week on average do you work out or exercise?" (N = 3530) and, "Did you exercise regularly in the last year?" (N = 907). Neighborhood social environment was measured by socioeconomic and social capital indicators. Neighborhood built environment was captured by pedestrian injury rate, residential density, distance to subway or parks, land use mix, and access to neighborhood amenities. ANALYSIS: Random effects logit and multinomial models. RESULTS: For weekly workout/exercise, individuals with access to restaurants and bars were more likely to report one to three times of weekly exercise (OR = 1.08; 95% CI: 0.99, 1.19) and four times or more weekly exercise (OR = 1.14; 95% CI: 1.03, 1.26) compared with those who reported no weekly exercise. For regular exercise in the past year, access to restaurants and bars (OR = 1.24; 95% CI: 1.05, 1.46) and neighborhood social environment (OR = 1.37; 95% CI: 1.11, 1.69) were significant. The social environment effects were stronger for women. CONCLUSIONS: Neighborhood social and built environments are both important for exercise independent of an individual's background. PMID- 19288847 TI - Neighborhood retail food environment and fruit and vegetable intake in a multiethnic urban population. AB - PURPOSE: To examine relationships between the neighborhood food environment and fruit and vegetable intake in a multiethnic urban population. DESIGN: Analysis of cross-sectional survey and observational data. SETTING: One hundred forty-six neighborhoods within three large geographic communities of Detroit, Michigan. SUBJECTS: Probability sample of 919 African-American, Latino, and white adults. MEASURES: The dependent variable was mean daily fruit and vegetable servings, as measured by using a modified Block 98 food frequency questionnaire. Independent variables included the neighborhood food environment: store availability (i.e., large grocery, specialty, convenience, liquor, small grocery), supermarket proximity (i.e., street-network distance to nearest chain grocer), and perceived and observed neighborhood fresh fruit and vegetable supply (i.e., availability, variety, quality, affordability). ANALYSIS: Weighted, multilevel regression. RESULTS: Presence of a large grocery store in the neighborhood was associated with, on average, 0.69 more daily fruit and vegetable servings in the full sample. Relationships between the food environment and fruit and vegetable intake did not differ between whites and African-Americans. However, Latinos, compared with African-Americans, who had a large grocery store in the neighborhood consumed 2.20 more daily servings of fruits and vegetables. Presence of a convenience store in the neighborhood was associated with 1.84 fewer daily fruit and vegetable servings among Latinos than among African-Americans. CONCLUSION: The neighborhood food environment influences fruit and vegetable intake, and the size of this relationship may vary for different racial/ethnic subpopulations. PMID- 19288848 TI - Modifiable health behaviors and short-term medical costs among health plan members. AB - PURPOSE: Describe the relationship among modifiable health behaviors and short term medical costs. DESIGN: Prospective study linking cross-sectional survey data that assessed modifiable risk behaviors with insurance claims. SETTING: A large health plan in Minnesota. SUBJECTS: A stratified, random sample of 10,000 yielded an analytic data set for 7983 members. MEASURES: The dependent variable was per member-per-month insurance payment plus subscriber liability. Eighteen months of medical costs were analyzed. Control variables included subscriber age, sex, type of insurance plan, days of enrollment, chronic disease status, education, and marital status. Independent variables, included self-reported health behaviors of smoking, heavy drinking, nutrition, and physical activity. ANALYSIS: Linear regression was performed on the natural log of the cost variable, followed by a retransformation to dollars. RESULTS: Physical inactivity and smoking were significant predictors of higher medical costs. Each day a member did not exercise there was a 2.9% difference in cost. Compared with never smokers, current smokers had 16% higher costs. Former smokers who had quit more than 1 year before taking the survey had 15% higher costs than never smokers. Recent former smokers cost 32% more than never smokers and more frequently experienced smoking-related medical conditions before they quit. Alcohol consumption was nonsignificant. Nutrition also was not significant but was narrowly measured by only fruit and vegetable consumption. CONCLUSION: Physical inactivity and smoking were associated with higher short-term medical costs among health plan members. PMID- 19288849 TI - Behavioral, demographic, psychosocial, and sociocultural concomitants of stage of change for physical activity behavior in a mixed-culture sample. AB - PURPOSE: Examine behavioral, demographic, psychosocial, and sociocultural concomitants of the stages of change for physical activity behavior among college students in South Korea (n = 221) and the United States (n = 166). METHODS: Measures obtained in this cross-sectional study included age; body mass index; nationality; gender; exercise behavior; processes of change; decisional balance; self-efficacy; stage of change; and predisposing, reinforcing, and enabling factors. RESULTS: The amount of variance explained for stage of change by the transtheoretical model constructs (i.e., decisional balance, processes of change, self-efficacy) ranged from 11% to 29% (all p < .001), whereas the predisposing (2%; p = .052), reinforcing (3%; p = .06), and enabling (5%; p < .001) factors were not as important. In multivariate ordinal logistic regression analysis, gender (odds ratio [OR] = 3.3; p < .001), gender by nationality interaction (OR = .27; p < .01), weekly exercise behavior (OR = 1.04; p < .001), and behavioral processes of change (OR = 1.12; p < .001) were each significant concomitants of the stages of change. DISCUSSION: In terms of physical activity behavior, South Korean women were more likely than South Korean men to be in the early stages, whereas American men were slightly more likely to be in the early stages than American women when all the concomitants were accounted for. Among the psychosocial stage of change concomitants, only the behavioral processes of change were found to be important. PMID- 19288850 TI - Clinical utility of a fingerstick technology to identify individuals with abnormal blood lipids and high-sensitivity C-reactive protein levels. AB - PURPOSE: The purpose of this study was to determine the ability of a commonly used fingerstick technology to identify individuals with abnormal blood levels of total cholesterol (TC), calculated low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), and high sensitivity C-reactive protein (hsCRP) compared with a standardized laboratory. METHODS: Participants (n = 250; mean age, 48.0 +/- 13.5 years; 66% female; 36% nonwhite) were eligible for primary prevention of cardiovascular disease (CVD). Blood lipids and hsCRP were measured simultaneously by (1) fingerstick analyzed by Cholestech LDX analyzers and (2) fresh venous blood that was analyzed by Columbia University General Clinical Research Center (GCRC) Core Laboratory. Pearson correlation coefficients, kappa, sensitivity, and specificity were calculated for fingerstick versus GCRC laboratory values for lipids and hsCRP. RESULTS: The correlations betweenfingerstick and core laboratory for TC, LDL-C, HDL-C, TG, and hsCRP were .91, .88, .77, .93, and .81, respectively (all p< .01). Sensitivity and specificity of the fingerstick to identify those with abnormal lipids and hsCRP > or = 1 mg/L were all > or = 75%. CONCLUSION: Fingerstick screening is accurate and has good clinical utility to identify persons with abnormal blood lipids and hsCRP at the point of care in a diverse population that is eligible for primary prevention of CVD. These results may not be generalizable to patients at high risk for CVD or who have known hyperlipidemia. PMID- 19288851 TI - State legislation to improve employee wellness. AB - OBJECTIVE: Categorize and describe the content and status of state legislation of worksite wellness. METHODS: State worksite wellness legislation was compiled from the Centers for Disease Control's Division of Nutrition, Physical Activity and Obesity State Legislative Database (http://apps.nccd.cdc.gov/DNPALeg/index.asp) and from LexisNexis (http://www.lexisnexis. com). Key word searches were used to gather worksite wellness legislation (2001-2006), with the exception of resolutions and those bills not pertaining to general employee wellness. Legislation was individually examined, categorized, and analyzed for content and status. RESULTS: The four categories of state legislation that appeared to be most common were tax credits (n = 34; 0 passed), wellness policies and programs (n = 21; 4 passed), alternative transportation (n = 18; 4 passed), and health insurance (n = 14; 3 passed). CONCLUSION: During 2001 to 2006, seven of 27 states enacted worksite wellness bills. In the three categories in which bills passed (wellness policies and programs, alternative transportation, and health insurance), 19% to 22% were enacted. This proportion, similar to other health promotions bills, indicates that worksite health promotion legislation passed as favorably as other health promotion topics. Further, the language in the bills did not recommend a specific standard for employee health, such as that in the national Healthy People 2010 objectives. PMID- 19288852 TI - Do employee health management programs work? AB - Current peer review literature clearly documents the economic return and Return on-Investment (ROI) for employee health management (EHM) programs. These EHM programs are defined as: health promotion, self-care, disease management, and case management programs. The evaluation literature for the sub-set of health promotion and disease management programs is examined in this article for specific evidence of the level of economic return in medical benefit cost reduction or avoidance. The article identifies the methodological challenges associated with determination of economic return for EHM programs and summarizes the findings from 23 articles that included 120 peer review study results. The article identifies the average ROI and percent health plan cost impact to be expected for both types of EHM programs, the expected time period for its occurrence, and caveats related to its measurement. PMID- 19288853 TI - [T serotypes distribution and antimicrobial susceptibility of Streptococcus pyogenes in children with pharyngotonsillitis in Asahikawa]. AB - Between June 2006 and April 2007, I measured T serotypes and antibiotic susceptibilities of 367 strains of Streptococcus pyogenes isolated from children with pharyngotonsillitis in Asahikawa. Prevalent serotypes were 12 (33.8%), 1 (22.9%), and 28 (12.5%). The MIC90s of beta-lactams were 0.008 microg/ml in penicillin G, cefcapene, cefditoren, cefteram, cefdinir and faropenem, and 0.015 microg/ml in amoxicillin. Of 367 isolates, macrolide-resistant (erythromycin > 0.5 micro/ml) strains account for 42 (11.4%). PMID- 19288854 TI - [Efficacy and safety of faropenem in pediatric patients with bacterial infectious diseases]. AB - The only oral penem antibiotic, faropenem (FRPM: Farom Dry Syrup for pediatrics), is one of the few antibiotics that exerts potent antibacterial activity against penicillin-resistant Streptococcus pneumoniae (PRSP), and the dosage and administration schedule has been established for children. We studied the efficacy and safety of the drug in 113 pediatric patients with mild-to-moderate bacterial infectious diseases: upper respiratory tract infection (pharyngitis or tonsillitis), acute bronchitis, otitis media and urinary tract infection (UTI). The patients were administered oral FRPM at the dose of 15-30 mg/kg/day three times a day for 3 to 8 days (or 5 to 14 days for group A streptococcal infection). The study drug was found to be clinically effective in 63/70 cases (90.0%) of upper respiratory tract infection, 6/7 cases of acute bronchitis, 16/17 cases (94.1%) of otitis media and 6/6 cases of UTI. FRPM was demonstrated to have very potent antibacterial activity against S. pneumoniae, with a high bacteriological eradication rate. No serious adverse drug reactions were observed. The only side effect was diarrhea in 12.5% of the patients (14/112 cases). There was little difference in the incidence of diarrhea between FRPM and other oral beta-lactam antibiotics. Compliance with FRPM was found to be very good in this study. These findings suggest that FRPM is as useful for the treatment of bacterial infectious diseases in children as oral penicillin and cephem antibiotics. PMID- 19288855 TI - Complement-mediated bactericidal effect of antibodies in human intravenous preparation against multi-drug resistant pseudomonas aeruginosa. AB - After incubation of four of ten strains of multi-drug resistant Pseudomonas aeruginosa (MDRP) with the healthy human serum for one hour, the bacteria lysed and the viable cell count decreased with the disappearance of the C3 complement. Over 99% of IgG was deleted from the serum by passing the serum through a Protein G Sepharose column. In such IgG-deleted serum, the bactericidal activity on P. aeruginosa was cancelled. The bactericidal activity recovered by adding human intravenous immunoglobulin preparation (IVIG) at the concentration of 2-4 mg/mL to the serum. Moreover, a strong bactericidal activity was shown in the case of some strains that the antibody titers of IVIG were high. It was thought that specific antibodies to some of P aeruginosa are contained in IVIG and activated the classical pathway of complement system; as a result, the bacterial lyses had been caused. Sera from healthy persons contain antibodies to P aeruginosa as well as complement components. Since the complement-mediated bactericidal activity disappeared by removing IgG in this study. These results suggest that the IVIG therapy in bacterial infections is expected to recover the immune function so as complement-mediated bactericidal activity as well as opsonic activity against P aeruginosa, especially in the patient with neutropenia. PMID- 19288856 TI - [Report of research grant: the Japan Antibiotics Research Association--Pfizer research fund on infectious diseases: Role of myeloid differentiation factor 88 in pneumonia induced by Escherichia coli or Streptococcus pneumoniae ]. PMID- 19288857 TI - [Lipid rafts controls the oncogenic potential of c-Src]. PMID- 19288858 TI - [Ion channels responsible for environmental response: molecular function of TRP channels activated by reactive chemical species]. PMID- 19288859 TI - [Imaging mass spectrometry: principle and application]. PMID- 19288860 TI - [Ribosomal RNA transcription and cellular energy control]. PMID- 19288861 TI - [Molecular mechanisms of ALS-linked mutant SOD1-induced motor neuron death]. PMID- 19288862 TI - [How is the catalytic pocket of the mature gamma-glutamyltranspeptidase formed upon autocatalytic processing of its precursor?]. PMID- 19288863 TI - [Creation of enzymes using computdtional de novo design: retro-aldol enzymes]. PMID- 19288864 TI - [Regulation of plant biomass production by transcription factors]. PMID- 19288865 TI - [Aureochrome: a blue-light receptor specific to stramenopiles]. PMID- 19288866 TI - [Inactivation of molecular function by light irradiation]. PMID- 19288868 TI - Enrollment explosion. Survey: 18% increase in health management programs. PMID- 19288867 TI - ER assistance requested. Congress eyes IT to pay for department pay bump. PMID- 19288869 TI - Profit or loss? PMID- 19288870 TI - Seeking alternatives. Providers take a variety of avenues in drive for capital. PMID- 19288871 TI - By the numbers. Largest for-profit hospital chains. PMID- 19288872 TI - Postcard: Beijing. PMID- 19288873 TI - A brief history of: abstinence. PMID- 19288874 TI - The trouble with repeat Cesareans. PMID- 19288876 TI - How to look young: eat more. PMID- 19288875 TI - Bunking in with mom and dad. PMID- 19288877 TI - Why bosses tend to be blowhards. PMID- 19288878 TI - Going nuts over nut allergies. PMID- 19288879 TI - Pocket-size personal trainers. PMID- 19288880 TI - Extracting a hefty profit. PMID- 19288881 TI - Dental implant research: a perspective. PMID- 19288882 TI - The use of laser technology (Er;Cr:YSGG) and stereolithography to aid in the placement of a subperiosteal implant: case study. AB - The use of laser technology has helped this clinician to provide treatment with less postoperative pain and increased healing. The subperiosteal implant is a modality that has been used for several decades, although its popularity has declined in favor of endosseous dental implants. In some instances, however, it remains the treatment of choice, specifically in the atrophic mandible (where placement of endosseous implants is not possible) or when placement would increase the chances of jaw fracture. This article reports the case of a patient rehabilitated using a simplified surgical protocol involving laser surgery and stereolithography. PMID- 19288883 TI - An implant-retained maxillary overdenture to obturate a patent oronasal communication: a case report. AB - This is a case report of dental prosthetic treatment of a 72-year-old patient with cleft palate with a persisting oral nasal communication in the anterior hard palate. The remaining teeth were extracted, and dental implants were surgically placed in the maxilla and mandible to retain removable overdentures. The maxillary complete overdenture had full palatal coverage as opposed to partial coverage. It was retained by 5 implants with low-profile light retentive retainers. The mandibular overdenture was retained with 4 implants of different diameters and brands. Apparent parameters for successful treatment of a removable maxillary overdenture are as follows: denture flanges, flat or lingualized occlusal scheme, light low-profile retainer retention, maximum implant positional spread with parallel placement, and a minimum of 4 rough-surfaced, long, wide implants placed in denser-type bone sites. PMID- 19288884 TI - Influence of interimplant distances and placement depth on papilla formation and crestal resorption: a clinical and radiographic study in dogs. AB - Among the factors that contribute to the papilla formation and crestal bone preservation between contiguous implants, this animal study clinically and radiographically evaluated the interimplant distances (IDs) of 2 and 3 mm and the placement depths of Morse cone connection implants restored with platform switch. Bilateral mandibular premolars of 6 dogs were extracted, and after 12 weeks, the implants were placed. Four experimental groups were constituted: subcrestally with ID of 2 mm (2 SCL) and 3 mm (3 SCL) and crestally with ID of 2 mm (2 CL) and 3 mm (3 CL). Metallic crowns were immediately installed with a distance of 3 mm between the contact point and the bone crest. Eight weeks later, clinical measurements were performed to evaluate papilla formation, and radiographic images were taken to analyze the crestal bone remodeling. The subcrestal groups achieved better levels of papillae formation when compared with the crestal groups, with a significant difference between the 3 SCL and 3 CL groups (P = .026). Radiographically, the crestal bone preservation was also better in the subcrestal groups, with statistically significant differences between the 2SCL and 2CL groups (P = .002) and between the 3SCL and 3CL groups (P = .008). With the present conditions, it could be concluded that subcrestal implant placement had a positive impact on papilla formation and crestal bone preservation, which could favor the esthetic of anterior regions. However, the IDs of 2 and 3 mm did not show significantly different results. PMID- 19288885 TI - Bone regeneration using beta-tricalcium phosphate in a calcium sulfate matrix. AB - The aim of the study was the histomorphometric comparison of the osteogenic potential of beta-tricalcium phosphate (beta-TCP) alone or in a calcium sulfate matrix. Three round defects, 10 mm (diameter) x 5 mm (depth), were created on each iliac crest of 4 dogs. The defects were divided into 3 groups. Ten defects were filled with beta-TCP in a calcium sulfate (CS) matrix (Fortoss Vital; group A), 10 defects were filled with beta-TCP alone (Fortoss Resorb; group B), and 4 defects were left ungrafted to heal spontaneously (group C). All defects were left to heal for 4 months without the use of a barrier membrane. Histologic evaluation and morphometric analysis of undecalcified slides was performed using the areas of regenerated bone and graft remnants. All sites exhibited uneventful healing. In group A sites (beta-TCP/CS), complete bone formation was observed in all specimens, graft granules dominated the area, and a thin bridge of cortical bone was covering the defect. Group B (beta-TCP) defects were partially filled with new bone, the graft particles still dominated the area, while the outer cortex was not restored. In the ungrafted sites (group C), incomplete new bone formation was observed. The outer dense cortical layer was restored in a lower level, near the base of the defect. The statistical analysis revealed that the mean percentage of new bone regeneration in group A was higher than in group B (49.38% and 40.31%, respectively). A statistically significant difference existed between the 2 groups. The beta-TCP/CS group exhibited significantly higher new bone regeneration according to a marginal probability value (P = .004 < .05). The use of beta-TCP in a CS matrix produced significantly more vital new bone fill and preserved bone dimensions compared with the use of beta-TCP alone. PMID- 19288886 TI - Early implant placement in bilateral sinus floor augmentation using iliac bone block grafts in severe maxillary atrophy: a clinical, histological, and radiographic case report. AB - Effectively restoring a grossly atrophic maxilla can be difficult for the implant surgeon. The placement of dental implants in patients who are edentulous in the posterior maxilla can present difficulties because of deficient posterior alveolar ridge and increased pneumatization of the maxillary sinus, resulting in a minimal hard tissue bed. Implant placement requires adequate quality and quantity of bone, especially in the posterior maxilla. Insufficient bone height and width in this area of the maxilla, because of expansion of the maxillary sinus and atrophic reduction of the alveolar ridge, represents a contraindication for conventional insertion of dental implants. The reconstruction of edentulous patients with adequate bone volume and density by the use of bone graft and, subsequently, the placement of dental implants has become a viable treatment option with high predictability. It is commonly shared that autologous bone graft is the gold standard grafting method in the augmentation of Higmoro antrum and in any kinds of guided bone regeneration. In this article, the authors report a case of severe maxillary atrophy that is augmented by block bone graft harvested from iliac crest. An early placement of implants is possible due to the quick healing of the site, as proven by histologic examinations. PMID- 19288887 TI - Patient-tailored medicine, part two: personalized medicine and the legal landscape. AB - In Part One, the authors addressed the relevance of genetic information, and how race and genetics have affected and may impact the development of medicines, pharmacogenomics, and personalized medicine in the United States. Part Two examines current and proposed federal and state laws and regulations intended to protect individuals from the misuse of genetic information, including uses that discriminate based on genetic predispositions. This Part next explores the potential for litigation against both manufacturers and providers,as well as potential defenses. The authors also discuss legal issues relating to research that relies on the use of genetic information. PMID- 19288888 TI - Patient access to unapproved therapies: the leading edge of medicine and law. AB - In 2007, the United States Court of Appeals for the District of Columbia rejected a claim that terminally ill patients have a constitutionally protected right to access experimental drugs that have passed Phase I testing, but have not yet received FDA approval. Since that case was decided, patients and patients' rights groups have continued to advance both constitutional and common law claims in an effort to compel FDA and manufacturers to grant access to unapproved therapies. Congress and FDA have also advanced proposals that would expand patient access to drugs that have not yet passed through Phase III trials. Whether patients with serious and life-threatening illnesses should be allowed to obtain experimental therapies before FDA approval is an issue that is poised to shape the practice of both law and medicine in the United States for some time to come. PMID- 19288889 TI - Enforcement related to off-label marketing and use of drugs and devices: where have we been and where are we going? AB - Off-label drug or medical device "use" is the practice of prescribing drugs or medical devices to patients for a purpose not included on the federally approved label. Off-label "marketing" is the practice of attempting to influence physicians to prescribe drugs or devices for off-label purposes. The federal Food and Drug Administration (FDA) maintains regulatory authority over the proper labeling of drugs and medical devices. Although not illegal, off-label use of certain drugs has led to controversy in recent years, especially in light of alleged behind-the-scenes marketing practices intended to increase off-label prescribing. Off-label marketing practices are prohibited and could result in criminal charges against a manufacturer, depending upon the circumstances. Yet a vast gray area exists for subtle marketing practices, such as circulating published medical studies about off-label uses to physicians. This article summarizes the legal and medical standards associated with off-label use and marketing of drugs, provides summaries of recent enforcement activities regarding off-label marketing, and explains the current federal regulatory issues surrounding off-label marketing practices. The authors provide practical pointers on regulatory compliance and the risks associated with fraud and abuse laws for drug companies and practitioners. PMID- 19288890 TI - Protecting yourself from your assertions: navigating multiple regulatory schemes and disclosure. AB - This article discusses the conflicting requirements of and assertions made to the Food and DrugAdministration 510(k) approval process and the United States Patent and Trademark Office patent application process for a medical device. Generally, in a 510(k) submission, a company asserts that its device is a substantial equivalent of a predicate device. Conversely, in a patent application, a company asserts that its device is new and nonobvious over all prior art devices. This article explores the risks that arise for a company, whether as a patent holder or an accused patent infringer, because of assertions made as part of a 510(k) submission. The authors look at the possibility of using a 510(k) submission against a company to invalidate that company's patent, then examine the possibility of using a company's 510(k) submission to show infringement of another company's patent. They discuss how regulatory and patent counsel can work together to navigate both regulatory schemes to minimize associated risks requiring credible arguments of substantial equivalence simultaneous with arguments of novelty and nonobviousness. PMID- 19288891 TI - A better approach to medical malpractice claims? The University of Michigan experience. AB - The root causes of medical malpractice claims are deeper and closer to home than most in the medical community care to admit. The University of Michigan Health System's experience suggests that a response by the medical community more directly aimed at what drives patients to call lawyers would more effectively reduce claims, without compromising meritorious defenses. More importantly, honest assessments of medical care give rise to clinical improvements that reduce patient injuries. Using a true case example, this article compares the traditional approach to claims with what is being done at the University of Michigan. The case example illustrates how an honest, principle-driven approach to claims is better for all those involved-the patient, the healthcare providers, the institution, future patients, and even the lawyers. PMID- 19288892 TI - Physician ownership and use of in-office advanced diagnostic imaging equipment: are IDTF standards a meaningful response to overutilization, quality, and costs? AB - The increase of physician ownership and use of in-office advanced diagnostic imaging equipment (magnetic resonance imaging, positron emission tomography, and computed axial tomography) has led to concerns about overuse, quality, and increased costs. The requirement that a physician or non-physician practitioner furnishing diagnostic testing services (1) enroll as an independent diagnostic testing facility (IDTF) for each practice location furnishing these services and (2) comply with the performance standards of 42 C.F.R. section 410.33 would represent a major step toward curbing potential advanced diagnostic imaging self referral overutilization, improving quality, and lowering costs. PMID- 19288893 TI - Hospital-physician joint ventures in a changing regulatory environment: planning for an unwind. PMID- 19288894 TI - [Acupuncture anesthesia' 50 years: beyond anesthesia and surgery]. AB - Today, when acupuncture and surgery have been combined for a half century, the author of the present paper holds that the main role of acupuncture in whole process of the surgery does not only mean anesthesia. Getting this point consciously, the author points out that acupuncture will not only have a bigger role in the operating room, but also plays an important and far-reaching impact on the acupuncturology and even the whole life science. PMID- 19288895 TI - [Effect of acupuncture on Cx43 knock-out mice dysmenorrhea response]. AB - OBJECTIVE: To observe the effect of acupuncture on dysmenorrhea response of Cx43 knock out mice. METHODS: Two-months-old heterozygote [Cx43(+/-)] type and wild type [Cx43(+/+)] female mice (20 mice/type) were allocated into normal group, model group, acupuncture group and Yimucao Gao (Extractum Leonuri Inspissatum) group randomly. Dysmenorrheal model was established by feeding the mouse with diethylstilbestrol (2 mg/g) once daily for 12 days. Ocytocin (i. p. 2 U/kg) was injected on the 12th day 1 h later after the feeding. Acupuncture of "Sanyinjiao" (SP 6) and "Diji" (SP 8) was given to mice of acupuncture group, once daily for 5 days from the 7th day on after modeling. The latency of body-writhing and the writhing number were measured. Ocytocin receptor (Oct-R) mRNA and vasopressin receptor (Vas-R) mRNA expression in the uterus tissue was detected by using reverse transcription polymerase chain reaction (RT-PCR) technique, and Oct-R expression was detected with immunohistochemistry after fixating and sectioning the uterus tissue. RESULTS: In comparison with their individual normal group, the latency of body-writhing of model groups in Cx43 (+/+) and Cx43 (+/-) mice shortened considerably, while the writhing number increased significantly (P<0.01). Compared with the corresponding model group, the writhing latencies of acupuncture and Yimucao Gao groups in Cx43 (+/+) mice and that of Yimucao Gao group in Cx43 (+/-) mice increased remarkably, and the writhing numbers of acupuncture and Yimucao Gao groups in Cx43 (+/+) mice and that of Yimucao Gao group in Cx43 (+/-) mice reduced markedly (P<0.05). In Cx43 (+/+) and Cx43 (+/-) mice, compared with their individual normal group, Oct-R and Vas-R mRNA expressions and Oct-R expression in model group were upregulated obviously (P<0.05). Compared with the corresponding model group, Oct-R and Vas-R mRNA expressions and Oct-R expression of acupuncture and Yimucao Gao groups in Cx43 (+/+) mice, and those of Yimucao Gao group in Cx43 (+/-) mice were down-regulated significantly (P<0.05). No significant differences were found between acupuncture and corresponding model groups in Cx43 (+/-) mice in the above-mentioned indexes (P>0.05). CONCLUSION: Acupuncture can inhibit diethylstilbestrol + ocytocin induced body writhing response in Cx43 (+/+) mice rather than in Cx43 (+/-) mice, which is closely related to its effects in down-regulating Oct-R mRNA, Vas-R mRNA and Oct-R expression. PMID- 19288896 TI - [Effect of electroacupuncture on the expression of corticotropin releasing hormone (CRH) and CRHR 1 mRNA in paraventricular nucleus of hypothalamus in stress-induced anxiety rats]. AB - OBJECTIVE: To investigate the effects of electroacupuncture (EA) on the expression of corticotropin releasing hormone (CRH) and CRHR 1 mRNA in paraventricular nucleus (PVN) of hypothalamus, to explore its mechanism in anti anxiety. METHODS: Thirty-three male SD rats were randomly divided into control, model and EA groups. Chronic emotional stress anxiety model was established by using chronic unpredictable emotional stress stimulation for 21 days. EA (15/25 Hz, 1-2 mA) was applied to "Baihui" (GV 20) and "Sanyinjiao" (SP 6) for 15 min/d and 21 days. The expression of CRH and CRHR 1 mRNA in hypothalamic PVN was measured by immunohistochemistry and in situ hybridization techniques separately. RESULTS: Compared with control group, the values of open-arms entries (OE%) and open-arms time (OT%) in model group were decreased significantly (P<0.01). In comparison with model group, OE% and OT% in EA group were increased obviously (P<0.05, P<0.01). Compared with control group, the optical density values of CRH immuno-reaction positive products and CRHR 1 mRNA expressed neurons of hypothalamic PVN in model group were increased obviously (P<0.05, P<0.01); while in comparison with model group, those of CRH and CRHR 1 mRNA in EA group were significantly decreased (P<0.05, P<0.01). CONCLUSION: EA can effectively relieve chronic stress stimulation induced anxiety in the rat, which is closely related to its effect in down regulating both CRH and CRHR 1 mRNA expression in PVN of hypothalamus. PMID- 19288897 TI - [Effect of electroacupuncture on apoptosis of hippocampus tissue in mice with vascular dementia]. AB - OBJECTIVE: To observe the effect of electroacupuncture (EA) on hippocampal apoptosis and learning-memory ability in vascular dementia (VD) mice so as to investigate its underlying mechanism in the treatment of VD patients. METHODS: Kunming mice were randomly divided into sham-operation (sham, n = 13), model (n = 15), EA (n = 16) and Nimodipine (intragastric gavage, 30 mg/kg for 15 days, n = 15) groups. VD model was duplicated by occlusion of bilateral carotid arteries and reperfusion. EA (2-80 Hz, 2 mA) was applied to "Dazhui" (GV 14), "Baihui" (GV 20), "Geshu" (BL 17) and "Zusanli" (ST 36) for 10 min, once daily for 15 days. Step-down and step-up tests were performed to assess the animal's memory and learning abilities separately; and the terminal deoxynucleotidyl transferase mediated dUTP-nick end-labeling (TUNEL) method was used to display the apoptotic cells of the hippocampus tissue. RESULTS: In comparison with sham group, the animals' reaction time upon electric shock stimulation, both step-up and step down error times, and hippocampal apoptosis number in model, EA and Nimodipine groups increased significantly (P<0.01), while the reaction latency upon electric shock of model, EA and Nimodipine groups decreased significantly (P<0.01). Compared with model group, the reaction time, both step-up and step-down error times and hippocampal apoptosis number of both EA and Nimodipine groups decreased significantly (P<0.05, P<0.01); while the reaction latency of EA and Nimodipine groups increased considerably (P<0.01). Comparison between EA and Nimodipine groups showed that the reaction latency of EA group was obviously longer than that of the later group (P<0.01), and the step-down error times of EA group was markedly lower than that of Nimodipine group (P<0.05). No significant differences were found between these two groups in other indexes (P>0.05). CONCLUSION: EA can ameliorate VD mice's learning-memory ability, which may be closely related to its effect in reducing hippocampal apoptosis. PMID- 19288898 TI - [Effect of electroacupuncture on caspase-9 expression in hippocampus and neuroethology in hyperlipemia rats with concurrent cerebral ischemia]. AB - OBJECTIVE: To observe the effect of electroacupuncture (EA) on hippocampal apoptosis protein caspase-9 expression and neuroethology in hyperlipemia plus cerebral ischemia (HL-CI) rats. METHODS: Seventy male SD rats were randomized into control, hyperlipemia (HL), CI, HL-CI, CI + EA, HL-CI + EA I , and HL-CI + EA II groups, with 10 cases in each. HL model was established by feeding the animals with high fat forage for 6 weeks and CI model was established by occlusion of the unilateral middle cerebral artery. EA (1-3 mA, 15 Hz) was applied to bilateral "Sanyinjiao" (SP 6) and "Fenglong" (ST 40) for 20 min every time; and "Baihui" (GV 20) and "Shuigou" (GV 26) were punctured and stimulated by twirling the acupuncture needle with hand continuously for 1 min. Acupuncture was given once daily for 17 days (beginning from the 10th day on before CI) in HL-CI + EA I group, and for 7 days (beginning after CI) in HL-CI + EA II group. The expression of hippocampal Caspase-9 was detected with immunohistochemistry. The score of neuroethology was also measured according to modified Bederson's method. RESULTS: In comparison with normal control and hyperlipemia model groups, Caspase 9 immune reaction (IR) positive cells in the hippocampus in HL-CI group increased significantly (P<0.01). After acupuncture Caspase-9 IR-positive cells decreased remarkably. In comparison with group HL-CI + EA II, Caspase-9 IR-positive cells decreased significantly in HL-CI + EA I (P<0.01). The score of neuroethology also degraded obviously. CONCLUSION: Acupuncture can improve neuroethology symptom, lessen over expression of hippocampal Caspase-9, and prevent CI injury in hyperlipemia rats with concurrent CI. PMID- 19288899 TI - [Effect of capsaicin-pretreatment on "Sibai" (ST 2)-electroacupuncture induced changes of c-fos expression in rat caudal spinal trigeminal nucleus]. AB - OBJECTIVE: To explore the afferent pathway of signals from electroacupuncture (EA) at "Sibai" (ST 2) acupoint. METHODS: A total of 36 SD rats were randomly divided into control, sham operation (sham), capsaicin, vehicle, capsaicin + EA, and vehicle + EA groups, with 6 cases in each. EA (1/20 Hz, 1-3 mA) was applied to "Sibai" (ST 2). After anesthesia (10% urethane), unilateral infraorbital nerve was separated to be immersed in a 1.5% capsaicin-fluid-soaked cotton ball for lesion of C fibers. Vehicle (20% tween) was used to immerse infraorbital nerve for vehicle and vechile + EA groups. The expression of c-fos in the caudal spinal trigeminal nucleus (cSTN) was detected by immunohistochemistry. RESULTS: In control group, c-fos immune-reaction (IR) positive neurons were observed to scatter in every layer of cSTN. No significant differences were found among vehicle, capsaicin, sham and control groups in the number of c-fos IR positive neurons in cSTN. The number of c-fos positive neurons in the I-II layers of cSTN was obviously increased following EA at ST2, being significant more than that in control, vehicle, capsaicin and sham groups (P<0.01 respectively). However, the number of c-fos IR-positive neurons in III-IV layers of cSTN had no marked change. In capsaicin + EA group, the number of c-fos-IR positive neurons in the I II layers of cSTN was obviously less than that in vehicle and vehicle + EA groups (P<0.01), but no significant changes were found in III-IV layers of cSTN. CONCLUSION: C afferent fibers of the infraorbital nerve are the primary afferent pathway of EA signals from Sibai (ST 2) stimulation in the rat. PMID- 19288900 TI - [Effect of electroacupuncture on inositol triphosphate and diacylglycerol levels in cerebral arteries of cerebral ischemia rats]. AB - OBJECTIVE: To observe the effect of electroacupuncture (EA) on inositol triphosphate (IP3) and diacylglycerol (DAG) contents in cerebral arteries of the cerebral cortex in rats with cerebral ischemia (CI), so as to explore its effect on intracellular signal transduction pathway. METHODS: Fifty-six Wistar rats wererandomly divided into control (n=8), CI (n=24) and EA (n=24) groups, and the later 2 groups were further separately divided into 1 h, 3 h and 6 h subgroups (CI-1 h, CI-3 h, CI-6 h, EA-CI-1 h, EA-CI-3 h and EA-CI-6 h), with 8 cases in each. CI model was made by occlusion of the middle cerebral artery. EA (15 Hz, 0.1 mA) was applied to "Shuigou" (GV 26) for 20 min. The cerebral arteries in the cerebral shallow layer were collected for assaying intracellular DAG content by thin-layer chromatography (TLC) and IP3 content by competitive protein binding assay (CPBA). RESULTS: Compared with control group,the levels of IP3 and DAG in cerebral vascular smooth muscle were significantly increased in model 1 h, 3 h and 6 h subgroups (P<0.01). Compared with the corresponding model subgroups,the levels of IP3 and DAG were significantly decreased in EA-CI-1 h, EA-CI-3 h and EA CI-6 h subgroups (P<0.01). CONCLUSION: EA of GV 26 can effectively inhibit CI induced increase of intracellular IP3 and DAG levels in rat cerebral arteries, which may contribute to its effect in modulating vascular motor to ameliorate cerebral microcirculation. PMID- 19288901 TI - [Effect of acupuncture on long-term potentiation of hippocampal CA 1 area in diabetic rats with concurrent cerebral ischemia]. AB - OBJECTIVE: To investigate the effect of manual acupuncture on long-term potentiation (LTP) in CA 1 area in hippocampal slices from rats suffered from diabetes mellitus (DM) and cerebral ischemia (CI). METHODS: A total of 54 female Wistar rats were randomly divided into 5 groups: normal control (Con), CI, DM + Sham-CI, DM + CI and DM + CI + acupuncture (Acup). DM model was established by i.p. of streptozotocin (60 mg/kg), and CI model duplicated by bilateral common carotid arteries occlusion and reperfusion. "Baihui" (GV 20), bilateral "Sanyinjiao" (SP 6) and "Pishu" (BL 20) or "Baihui" (GV 20), bilateral "Shenshu" (BL 23) and "Zusanli" (ST 36) were punctured alternatively and stimulated manually, once every other day for 15 times. Hippocampal slices were taken 40 days after modeling. LTP in hippocampal CA 1 induced by high frequency stimulation (100 Hz, 100 pulses) of Schaffer collaterals and population spikes (PS) were recorded by using glass micropipettes. RESULTS: No significant differences were found between DM + CI and normal control groups in the shape and amplitude of PS (P>0.05). After stimulation of Schaffer collaterals, the amplitude and slope rate of PS in hippocampal CA 1 of DM + CI rats were decreased significantly in comparison with normal control group (P<0.05, P<0.01). While compared with DM + CI group, both the amplitude and slope rate of PS in DM + CI + Acup group increased significantly (P<0.05, P<0.01), suggesting basic restoration of LTP after acupuncture. CONCLUSION: Acupuncture can promote the recovery of DM+CI induced decline of the amplitude and slope rate of PS, which may contribute to its effect in improving memory ability in DM + CI rats through regulating hippocampal neuronal synaptic plasticity. PMID- 19288902 TI - [Effect of acupuncture on partial oxygen pressure of deep tissue along the Governor Vessel in 31 normal volunteer subjects]. AB - OBJECTIVE: To observe changes of the partial oxygen pressure (PO2) of deep tissues along the Governor Vessel and during acupuncture, so as to probe into the characteristics of energy metabolism in the deep tissues along the meridian track. METHODS: Thirty-one healthy volunteers were enrolled in the present study. PO2 values in Zhongshu (GV 7), Zhiyang (GV 9) and Mingmen (GV 4) of the Governor Vessel, and 2 non-acupoints (below the 4th and the 12th vertebrae respectively) were detected by using a three-channels needle-type sensor inserted into the tissues about 1.5 cm deep under the skin. The detection was conducted continuously for 60 minutes before, during and after electroacupuncture (EA, 2 Hz, 20 min) stimulation of Yaoyangguan (GV 3). RESULTS: At normal state (before EA), PO2 values were (10.98 +/- 5.81) kPa in Governor Vessel (GV 4, GV 7 and GV 9), being significantly higher than those [(5.00 +/- 3.88) kPa, (4.74 +/- 4.23) kPa] of the left and the right control points separately. During acupuncture of GV3, PO2 value [(9.36 +/- 5.99) kPa] in Governor Vessel decreased significantly in comparison with that of pre-EA; while those [(5.83 +/- 4.75) kPa and (5.58 +/- 15.28) kPa] in the left and the right control points had no obvious changes (P>0.05). And after acupuncture, the reading of PO2 was (8.62 +/- 5.11) kPa along the Governor Vessel being comparable to that of EA-period, and those [(5.74 +/- 4.76) kPa and (5.61 +/- 5.45) kPa] in the left and the right control points still had no obvious changes. CONCLUSION: PO2 in the deep tissues along the Governor Vessel is significantly higher than those of its bilateral control points, and decreases during and after EA, displaying a more vigorous energy metabolism in the deep tissues along the Governor Vessel in normal state, and up-regulation after EA. PMID- 19288903 TI - [Randomized control study on the treatment of 26 cases of acne conglobata with encircling acupuncture combined with venesection and cupping]. AB - OBJECTIVE: To observe the curative effect of acne conglobata treated by encircling acupuncture combined with ventouse and cupping. METHODS: A total of 52 acne conglobata patients were randomly divided into acupuncture group (n=26) and Western medicine group (n=26). Patients of acupuncture group were treated with encircling acupuncture around the affected focus. Common acupuncture was applied to Hegu (LI 4), Xuehai (SP 10), Fenglong (ST 40) and Sanyinjiao (SP 6), once daily. Dazhui (GV 14) and Feishu (BL 13) were used for venesection and cupping (twice a week). Patients of medication group were treated with oral administration of Isotretinoin Capsules (10 mg, t.i.d.). The treatment duration of 2 groups was 4 weeks. Serum IL-6 content was detected with double-antibody sandwich elisa enzyme linked immunosorbent assay (ELISA). RESULTS: After the treatment, in acupuncture group and Western medicine group, 3 (11.5%) and 4 (15.4%) cases experienced remarkable relief in their signs, 14 (53.8%) and 11 (42.3%) had marked improvement, 6 (23. 1%) and 7 (26.9%) had improvement, 3 (11.5%) and 4 (15.4%) failed, with the effective rates being 88.5% and 84.6%, respectively. No significant difference was found between two groups in the therapeutic effect (P>0.05). Self-comparison showed that after the treatment, IL 6 in both groups decreased significantly (P<0.01). The therapeutic effect of acupuncture group was significantly superior to that of Western medicine group in lowering serum IL-6 (P<0.05). CONCLUSION: Both acupuncture and medication can effectively promote the recovery of the affected skin, and lower serum IL-6 level in acne conglobata patients. The effect of acupuncture is stronger than that of Isotretinoin Capsules in lowering serum IL-6 content and has fewer adverse effects. PMID- 19288904 TI - [Influence of ginger-partitioned moxibustion on serum NO and plasma endothelin-1 contents in patients with primary dysmenorrhea of cold-damp stagnation type]. AB - OBJECTIVE: To explore the mechanism of ginger-partitioned moxibustion in the treatment of cold-damp stagnation type primary dysmenorrhea (PD) patients. METHODS: A total of 209 PD outpatients of cold-damp stagnation type from 3 hospitals were randomized into moxibustion group (n = 105) who were treated with ginger-partitioned moxibustion, and control group (n = 104) who were asked to take Yueyue Shu Granules (an effective patent drug for PD). Ginger-partitioned moxibustion was applied to Shenque (CV 8) and Guanyuan (CV 4) from the menstrual onset on for the first course and 3 days before the onset for the second and third course, continuously for 3 days. Before and after the treatment, plasma endothelin-1 (ET-1) and serum nitric oxide (NO) contents in those patients (n = 40/group) and normal women (n = 20) experiencing menstrual period were determined by radioimmunoassay and nitrate reductase method. RESULTS: After the treatment, of the 105 and 104 cases in moxibustion and control groups, 58 and 32 cured, 37 and 33 markedly effective, 5 and 24 effective, 5 and 15 failed, with the effective rates being 95.24% and 85.58% respectively. The therapeutic effect of moxibustion group was significantly better than that of control group (P<0.05). Before treatment, in comparison with normal group, plasma ET-1 contents in both moxibustion and control groups were significantly higher (P<0.01), while serum NO contents in these two groups markedly lower (P<0.01). After the treatment, self comparison of both moxibustion and control groups showed that plasma ET-1 decreased significantly and serum NO levels in increased considerably (P<0.05, P<0.01), and the therapeutic effect of moxibustion group was markedly superior than that of control group in lowering plasma ET-1 (P<0.01). CONCLUSION: Ginger partitioned moxibustion is effective in relieving primary dysmenorrhea patients' abdominal pain, which may be closely associated with its actions in lowering plasma ET-1 level and raising serum NO contents. PMID- 19288905 TI - [Systems biology and modernization of acu-moxibustion medicine]. AB - Systems biology, a major science characterized by holistic view, will give a far reaching impact on our understanding about the essence of biosystem. In the present study, the authors analyze the methodological characters of the systems biology, its relation to and its possible impact on acu-moxibustion medicine. Up to now, modern studies have revealed only partial content of traditional Chinese medicine including acu-moxibustion medicine. With the help of methods, techniques and research thoughts of systems biology, the mechanism of basic theory and clinical efficacy of acupuncture and moxibustion will be understood thoroughly and more rapidly. PMID- 19288906 TI - [Advances of clinical studies on acupuncture treatment of depression]. AB - An increasing evidence shows that in the treatment of depression, acupuncture therapy is definitely effective. It has fewer side-effects and is extensively applicable for different types of patients. In the present paper, the authors follow the methodology of evidence-based medicine to make a systemic summary about acupuncture treatment of depression from 1) different therapeutic methods, such as electroacupuncture, acupuncture combined with psychological comforts, acupuncture combined with medication, etc. 2) different states of disease, and 3) special disease-induced psychological disorders. Moreover, the authors also make a discussion and a prospect on the treatment of depression and put forward some suggestions for clinical researches in the coming days. PMID- 19288907 TI - [Anatomical visibility of the meridian information channels]. AB - The "structure imperfectness of signal channel rule" put forward by the author of the present paper may provide a theoretical evidence for the systematicness of meridian information channel. The conclusion that no special structure of the meridian-collateral system has been found is likely to serve as a piece of counterevidence. According to the latest structural view, the development of capillaries, lymphatic vessels and nerves needs target cells-released inducible factors. In the initial phase of the development of the organism, the asymmetry of the interspaces among cells results in the production and arrangement imbalance of the sequential factors which make the capillaries, lymphatic vessels and nerves distribute sequentially in time and space. Meridian-collateral, following the "systemic statistic distribution rule", is a general expression of this distribution pattern. As a systematic structure, the meridian-collateral system distributes in an optimized way in the human body and has both orderly and compatible characteristics. The author thinks that the meridian-collateral information channel is anatomically visible in the time and spatial structure, and in its logical structure and compatibility during the process of growth. Hence, many techniques of delicate anatomy, quantitative anatomy, growth anatomy, comparative anatomy, tridimensional remodeling of living creature and dynamical remodeling of growth all should be used as the important tools for studying the meridian information channel. The theory and the anatomical techniques determine what you would finally find. PMID- 19288908 TI - [Malaria situation in the People's Republic Of China in 2007]. AB - Total 50148 malaria cases and 83551 suspected cases with 18 deaths were reported by the annual case reporting system in 1182 counties of 23 Provinces/Municipality/Autonomous Region (P/M/A) in 2007, and the annual incidence was 0.39/10000. Through the internet reporting system 46,988 malaria cases were reported from 1097 counties of 31 P/M/A. The number of malaria cases and the rank of P/M/A were basically in concordance in two systems. Among the 1182 counties with reported malaria cases, 26 counties with an incidence of more than 10/10000 distributed in Yunnan (9 counties), Hainan (5), Anhui (9), Henan (2), and Tibet (1). There were 72 counties in which the malaria incidence was between 1/10000 and 10/10000. 1830 Plasmodium falciparum malaria cases accounted for 3.6% of the total cases, of which 65.1% (1192) were imported cases reported in 187 counties/cities of 18 P/M/A. Indigenous falciparum malaria was found in 37 counties/cities of Yunnan and Hainan Provinces, of which 24 counties/cities were in Yunnan, decreased by 2, 13 counties/cities were in Hainan, 1 increased compared to that of 2006. Focal outbreaks occurred in 210 villages of 10 counties in Yunnan, Guizhou, Henan and Hubei provinces. 679 malaria cases resulting from outbreaks accounted for 1.4% of the total reported cases. Although a considerable decrease in malaria incidence contributed to the implementation of the National Malaria Control Program and a project supported by the Global Fund, Yunnan and Hainan Provinces were still the relatively high transmission areas. Yunnan ranked No.2 in the country in terms of the number of cases, while Hainan ranked down to No.2 by malaria incidence since 2006. 13157 malaria cases were reported from the two provinces in 2007, accounting for 26.2% of the total reported cases in the country. There were 9770 cases with 9 deaths reported from Yunnan, the incidence was 2.44/10000, a decrease of 37.3% than that in the last year. Among the reported cases, 1486 were falciparum malaria with 65.6% imported cases. The number of reported cases in Hainan was 3 387, with an incidence of 4.05/10000, 12.9% decrease than the last year. In central China, the re-emergence of malaria was held back on the whole in 2007, Anhui was still the number one in the country by either the number of cases or malaria incidence. 27 278 malaria cases were reported from Anhui Province in 2007, accounting for 54.4% of the total cases in the country, with an incidence of 5.0/10000 decreased by 21.9% than that in 2006. The number of reported cases in Henan Province was 4174, decreased by 21.4% in incidence. Hubei Province reported 1768 malaria cases with an incidence of 0.31/10000, same as the 2006. 940 cases were reported from Jiangsu Province, increased by 22.6% compared to that in 2006. Cases reported from other P/M/A occupied about 5.7% of the total. Several hundreds were reported from each of Guizhou, Sichuan, Guangdong, Zhejiang, Shandong and Shanghai. Less than 100 cases were reported from each of Guangxi, Fujian, Jiangxi, Hunan, Chongqing, Liaoning, Shaanxi, Shanxi, Gansu Provinces in 2007. In summary, the re-emergence of malaria has been basically contained through several years' effort, but it is still an important problem of public health in China, especially in the southern and central parts. Yunnan and Hainan still faced a severe situation of malaria endemic with the spread of Plasmodium falciparum, especially imported malaria in the border areas. In the central parts of the country, especially in Anhui, malaria was considerably serious with the highest incidence and most malaria cases in 2007. In addition, provinces not covered by the Global Fund Program such as Guizhou, Zhejiang, Guangdong were confronted an increasing disease burden by malaria, which may become a new challenge to the National Malaria Control Program. PMID- 19288909 TI - [Discovery of a residual focus of bancroftian filariasis after declaration of its transmission interruption in Guangxi]. AB - OBJECTIVE: To describe the discovery of a residual foci of bancroftian filariasis in Fuchuan County where the disease was announced to have been eliminated, and reveal its epidemiologic feature. METHODS: The investigation was carried out from August 2007 to March 2008 among residents in Changtang village where the first case of filariasis was found and the neighboring villages. They were screened with two thick blood smears. Immunochromatographic technology (ICT) was conducted for those going out but returned and those in surrounding areas. Vector mosquitoes were collected and dissected to find filaria larvae. Historical documents were reviewed and relevant people were interviewed. RESULTS: In Changtang administrative village, 1052 residents were screened and 19 cases with microfilaremia were found in 2 natural villages, with a Mf-positive rate of 1.8% (5.1% in Gangshang and 1.4% in Yinshan respectively). No Mf-positive case was found in 4119 residents screened in other 3 villages. The average microfilaria density in the 19 cases was 17.37/60 +/- 1 blood. All the 19 cases belonged to 12 families, and 13 cases were relatives to each other, which showed a feature of spatial clustering and family clustering. More patients were identified in the age groups of 20-29 and 50-59, and 57.9% of them were older than 50 years. No larvae were found in 54 Culex pipiens fatigans dissected. CONCLUSION: The Changtang village is identified as a residual focus of bancroftian filariasis with a low, limited endemicity. More cases have been among the elderly with a low average microfilaremia. PMID- 19288910 TI - [A case of falciparum malaria complicated with dengue fever]. PMID- 19288911 TI - [Follow-up survey on the imported cases of lymphatic filariasis in Guangdong Province]. AB - OBJECTIVE: To make a survey on the possible imported cases with microfilaremia from a recently identified outbreak focus in Fuchuan County of Guangxi Region. METHODS: Moving people as temporary laborers from Guangxi and periphery residents were screened by thick blood smears for microfilariae and ELISA for specific IgG4 in Sept 2007. RESULTS: 989 persons in cities/districts of Nansha, Panyu, Shantou and Zhongshan were investigated. Among 150 persons from Changtang administrative village of Guangxi, 47 were from two outbreak focuses, Ganshang and Yinshan natural villages in Changtang. Three cases with microfilaremia were found, with a microfilaraemia density of 24, 20 and 2 per 120 microl blood, respectively. The 3 cases were all from the 2 natural villages, with a microfilaraemia positive rate of 6.4% (3/47). Meanwhile, anti-filaria IgG4 was detected for other 1 501 persons, including those from the outbreak focuses, from surr-founding areas and from non-endemic area as control, fifteen cases were found with IgG4 positive. Of the 15 positive cases, 12 were from the 2 outbreak natural villages, with a positive rate of 25.5%, higher than those of others (chi2=295.83, P < 0.01). The cases with IgG4 positive but microfilaremia negative were reexamined by ICT kit and all showed negative. CONCLUSION: It is suggested that no further transmission occurs although microfilaremia cases have been imported from the outbreak focus of Guangxi. PMID- 19288912 TI - [A multi-gene DNA vaccine encoding non-fusion membrane-anchoring antigen against Schistosoma japonicum]. AB - OBJECTIVE: To construct DNA vaccine (pIRES-Sj97-Sj14-Sj26) and study its immunogenicity and protective immunity against Schistosoma japonicum. METHODS: The plasmid pIRES-Sj97-Sj14-Sj26 containing fatty binding protein (Sj14), GST (Sj26) and paramyosin (Sj97) was constructed and expressed on the membrane. RT PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, and IFA for detecting the expression of trans-membrane Sj14, Sj26 and Sj97. Sixty BALB/c mice were randomly divided into 3 groups. Mice in each group respectively received normal saline, pIRES blank vector and pIRES-Sj97-Sj14 Sj26 by intramuscular injection. Two weeks after the 3rd immunization, 10 mice from each group were sacrificed and total IgG in serum and the level of IFN-y were detected by ELISA and lymphocyte stimulating index (SI) by MTt. FCM was used to analyze the subgroups of splenocytes. The level of NO secreted by peritoneal macrophages was determined by nitrate reductase approaches. The left 10 mice in each group were challenged with (40 +/- 1) cercariae of S. japonicum by abdominal skin penetration. Forty-five days after challenge, mice were sacrificed, and numbers of recovered worms and hepatic eggs were counted. RESULTS: RT-PCR showed the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA. IFA proved the expression of Sj26, Sj14 and Sj97 protein. Level of total IgG in the vaccination group, saline group and pIRES blank vector group was (5.62 +/- 0.64), (1.22 +/- 0.20) and (1.48 +/- 0.36) mg/ml respectively, showing a statistical significance (P < 0.01, P < 0.05). The NO level in macrophages was (321.19 +/- 18.03), (184.12 +/- 11.05) and (213.51 +/- 15.93) nmol/ml in the 3 groups respectively (P < 0.05), and the lymphocyte stimulating index in the 3 groups was (2.25 +/- 0.29), (1.18 +/- 0.07) and(1.22 +/- 0.09) respectively (P < 0.01). The INF-gamma level was higher in the vaccination group than others (P < 0.01). The percentage of CD4+ and CD8+ T cells increased considerably (P < 0.01). The vaccination group showed a worm reduction rate of 39.9% (P < 0.01) and an egg reduction rate of 43.9% (P < 0.01). CONCLUSION: The vaccine candidate pIRES-Sj97-Sj14-Sj26 induces an immune protection in BALB/c mice against Schistosoma japonicum. PMID- 19288913 TI - [Evaluation on malaria situation in areas along Yellow River and Huaihe River by indirect fluorescent antibody test]. AB - OBJECTIVE: To evaluate malaria situation by indirect fluorescent antibody test (IFAT) in 12 villages of six counties along Yellow River and Huihe River in Anhui, Henan and Hubei Provinces. METHODS: Cross sectional seroepidemiologic survey was conducted through IFAT in Anopheles sinensis area and A. anthropophagus plus A. sinensis area respectively in 2006 and 2007. The blood filter papers collected from residents were tested by IFAT to calculate malaria antibody positive rate (APR) and positive GMRT in different age groups. The predictive infection probability was evaluated by reversible catalytic model based on antibody positive rate in the group of less than 20 year-old and compared to the annual parasite incidence. RESULTS: In 2006 and 2007, the average antibody positive rate in the investigated villages was 6.1% and 12.0% respectively. In An. sinensis areas, the APR in 2007 (12.0%) was significantly higher than that in 2006 (4.1%) (chi2 = 69.9, P < 0.01) while in An. anthropophagus plus An. sinensis areas there was no significant difference in 2006 (10.3%) and 2007 (12.1%) (chi2 = 0.17, P > 0.05). The average positive GMRT of the 12 villages was 26.2 in 2007, slightly higher than that in 2006. In both An. sinensis area and An. anthropophagus plus An. sinensis area, the predictive infection probabilities were higher than the annual parasite incidence with an average 117.3 and 17.20 times higher than the latter respectively. 87.8% of the antibody positive cases were asymptomatic, indicating that there were potential infection sources with symptomatic parasitaemia in the areas. CONCLUSION: Malaria is still prevalent in areas along Yellow River and Huihe River and the actual number of malaria cases are underestimated by the case report system. Malaria reemergence may occur in An. sinensis area but relatively stable in An. anthropophagus plus An. sinensis area. PMID- 19288914 TI - [One case of human echinostomiasis in Ganzhou]. PMID- 19288915 TI - [Urticaria in relation to mite sensitivity and immunotherapy with Injection dermatophagoidei farinae]. AB - OBJECTIVE: To investigate the prevalence of mite sensitivity in patients with urticaria or other skin rashes, and to observe the clinical efficacy of a specific immunotherapy (SIT) by the Injection dermatophagoides farinae for the patients. METHODS: In 7-year period (1998-2005), skin prick test (SPT) with a dust mite (Df) allergen was carried out to detect the prevalence of mite sensitivity in OPD patients suffering from skin rashes. Among the patients sensitive to mite with SPT > or =++ response, 3 groups were established. In group A, routine SIT with Injection dermatophagoides farinae was conducted. In 9-week increasing dose phase, three stepwise increasing volumes (0.3 ml, 0.6 ml and 1.0 ml) each case was injected subcutaneously with mite concentration of 1:100000 (w/v), 1:10000 (w/v) or 1:5000 (w/v) respectively once a week, followed by a maintenance dose phase for an injection with 1:5000 (w/v) 1.0 ml/wk for 6 weeks. Group B received rush SIT with mite injections. A total of 15 injections in a course of therapy with same concentration and volume was given as those for the routine ones except shortened intervals, namely, 9 initial injections completed in 3 days by three injections of each concentration per day with two 30 min intervals, maintenance doses were then provided in 6 days with 1:5000 (w/v) 1.0 ml/d. Thereafter, both groups A and B were maintained for one year with a dose of 1:5000 (w/v) 1.0 ml every 2 wk. Group C received antihistamine treatment as control, the patients received daily oral Ebastine 10 mg in the morning and Cetirizine dihydrochloride 10 mg in the evening for one week course and pro re nata later. Levels of serum tIgE and serum mite sIgE were detected by ELISA in 20 urticaria cases before and after one year mite SIT. RESULTS: Altogether, 2685 cases with skin rashes were detected by Df allergen SPT. The prevalence of urticaria cases sensitive to mite was 70.3% (1754/2496), which was higher than that of eczema 63.5% (54/85) and anaphylactoid purpura 60.6% (63/104) (P < 0.05) . 248 cases of urticaria sensitive to mite with SPT > or = ++ response received SIT with Injection dermatophagoides farinae for one year, clinical evaluation revealed an overall efficacy of 91.1% (226/248) with 66.1% (164/248) of excellent or good results, significantly higher than that of antihistamine treatment [12.7% (20/158)] (P < 0.01). Faster improvement of clinical symptoms was shown in rush SIT (group B) than that of routine one (group A), with higher efficacy in group B than group A (excellent and good results being 76.7% and 55.0% respectively) (P < 0.05). Serum tIgE and mite sIgE in 20 urticaria cases were detected before and after one year mite SIT, showing that tIgE decreased by half in 40% (8) of the patients, while serum mite sIgE level increased significantly (P < 0.01) one year later. CONCLUSION: Mite allergen SPT is an etiological diagnostic technique for urticaria patients sensitive to mite. Clinical efficacy of mite allergen SIT has been proved to be good for the patients, and the rush SIT shows quicker effect of relieving symptoms and better efficacy than that of the routine immunotherapy. PMID- 19288916 TI - [Suppression effect of different stage antigens of Schistosoma japonicum on airway inflammation in a murine model of asthma]. AB - OBJECTIVE: To investigate the effect of antigens of different stage Schistosoma japonicum on airway inflammation in a murine model of asthma. METHODS: 48 female BALB/c mice were randomly divided into eight groups. Mice in group A were given normal saline of equal volume as control. Group B was asthma model which was established by intraperitoneal and intranasal challenge with OVA. Mice in groups C, D and E were immunized with soluble egg antigen (SEA), soluble male worm antigen (SWA), and schistosomulum antigen (SSA) respectively 4 times in a week interval, followed by OVA sensitization as in group B 1 week after the final immunization. Mice in groups F, G, and H were immunized with SEA, SWA, and SSA respectively but sensitized and challenged with saline instead of OVA. 48 hours after asthma was induced, the mice were sacrificed. Leukocytes and eosinophils were counted in bronchoalveolar lavage fluid (BALF). The level of IL-5, IL-10 and IFN-gamma in BALF was detected. Pathologic changes in lung tissues were observed. RESULTS: Inflammation cells, especially eosinophils, appeared in airways of mice in groups B, C, D and E, but with much less number in groups C, D and E. No inflammation cells were seen in airways of group A mice. The number of leukocytes, eosinophils and level of IL-5 in BALF of group B [(98.4 +/- 16.1) x 10(4)/ml, (17.6 +/- 4.3) x 10(4)/ml, (197.9 +/- 36.5) pg/ml respectively] were significantly higher than those of group A [(8.2 +/- 1.1) x 10(4)/ml, (0.02 +/- 0.01) x 10(4)/ ml, (12.3 +/- 7.4) pg/ml], however the levels of IL-10 and IFN gamma were significantly lower than that of group A (P < 0.05). The number of leukocytes, especially eosinophils, in BALF of groups C, D and E was significantly lower than that of group B. The level of IL-5 in BALF of groups C, D and E was significantly reduced, while that of IL-10 and IFN-gamma in BALF of the 3 groups was significantly higher than group B (P < 0.05). CONCLUSIONS: The immunization with S. japonicum antigens can effectively modulate the level of cytokines and inhibit the eosinophil infiltration and airway inflammation in asthmatic mice. PMID- 19288917 TI - [Expression of multiepitope antigenic gene of Toxoplasma gondii in transgenic tomatoes]. AB - OBJECTIVE: To obtain the transgenic tomato plant expressed multiepitope antigenic gene of Toxoplasma gondii (Tg-MAG). METHODS: Tg-MAG under the control of E35S promoter, E35S-E81.1 chimeric promoter and E8 2.2 promoter in the plant expression vectors was transferred into the tomato cotyledons and hypocotyls via Agrobacterium-mediated T-DNA transformation. The candidate buds were obtained and elongated by kanamycin resistant screening. After further rooting selection, the selected tomato lines were trained and transplanted to soil and grown in a greenhouse till blossoming and bearing fruit. The transgenic tomato plants were identified using PCR, RT-PCR and Western blotting. RESULTS: Tg-MAG was introduced into the transgenic plant genomic DNA and transcribed correctly at expected size 360 bp identified by PCR and RT-PCR. Western blotting results indicated that the recombinant Tg-MAG from 8 transgenic lines, expressed in tomato fruits at expected size 11900, 2 of them showed strong reaction band with HRP labeled McAb K7H3 against the major surface antigen 1( SAG1) of T. gondii tachyzoites. CONCLUSION: Tg-MAG transgenic tomato lines were obtained. Tg-MAG recombinant protein with good immune activity was expressed successfully in transgenic tomato fruits. PMID- 19288918 TI - [In vitro effect of (+)-usnic acid on Toxoplasma gondii tachyzoites]. AB - OBJECTIVE: To explore the effect of usnic acid on Toxoplasma gondii tachyzoites in vitro. METHODS: There are four groups named as (+)-usnic acid group, acetylspiramycin group, DMSO group and normal saline group. Groups of (+)-usnic acid and acetylspiramycin were further divided into 4 subgroups with final concentration of 5, 10, 25, 50 microg/ml respectively. Normal saline group and DMSO group were respectively given equal volume normal saline and 1% DMSO. Each group have 15 parallel tubes with 1 ml (1 x 10(6)/ml) T. gondii tachyzoites aqueous suspension. At 1 h, 2h and 4 h after drug treatment, tachyzoites were counted by light microscope with 0.4% Trypan blue staining. Tachyzoites in aqueous suspension was collected, and washed 3 times by PBS solution. Normal mice were inoculated intraperitoneally and observed for three generations. The cultivated rat cardiofibroblasts were then infected in vitro with T. gondii tachyzoites. At the same time, rat cardiomyocytes invasion by T. gondii tachyzoites was investigated. RESULTS: At 4 h treated by 10, 25 and 50 microg/ml (+)-usnic acid, 100% T. gondii tachyzoites were stained. Some tachyzoites were swelling, blunt or round in the two ends; and granules appeared in the cytoplasm, the nuclei were deep stained. The changes of tachyzoites in acetylspiramycin group were similar to (+)-usnic acid group, 100% T. gondii tachyzoites were stained in 50 microg/ml acetylspiramycin subgroup. In inoculation tests, mice died at 8th to 9th days in 5 microg/ml (+)-usnic acid subgroup and numerous tachyzoites were detected in ascites. However, most mice survived to be killed in the other (+)-usnic acid subgroups and the tachyzoites were not found in ascites. All mice in acetylespirmycin groups died at 6th to 8th days after inoculation and many tachyzoites or pseudocysts were observed in mice ascites. In infecting cell tests, the cultivated rat cardiofibroblasts were infected in vitro by the tachyzoites after treated with 5 microg/ml (+)-usnic acid for 4 h, and pseudocysts were formed in infected cells. It was negative in the other subgroups of (+)-usnic acid. But the cultivated rat cardiofibroblasts were infected to varying degree in acetylspiramycin groups, normal saline group and DMSO group. CONCLUSION: (+)-Usnic acid has a remarkable effect on T. gondii tachyzoites. PMID- 19288919 TI - [Therapeutic efficacy of naphthoquine phosphate combined with artemisinine against Plasmodium knowlesi]. AB - OBJECTIVE: To study the antimalarial activity of naphthoquine phosphate combined with artemisinine against Plasmodium knowlesi in rhesus monkey. METHODS: Monkeys were randomly divided into 9 groups (3/group). The monkeys in groups A and B were treated i.g. once daily for 3 days with 6 or 10 mg/kg of naphthoquine phosphate respectively. Those in groups C and D were treated i.g. twice for the 1st day and once for the 2nd and 3rd day with 31.6 or 100 mg/kg of artemisinine respectively. In groups E, F and G, they were treated i.g. only once with the combination of naphthoquine phosphate 10 mg/kg and artemisinine 10, 20 or 25 mg/kg respectively. Groups H and I served as controls which were treated i.g. only once with 10 mg/kg of naphthoquine phosphate and 30 mg/kg of artemisinine respectively. Parasitemia was examined beginning 24 h after drug administration. The observation lasted 105 days when no more parasite was found. RESULTS: At 24 h after drug administration, the parasite reduction rate in all groups was higher than 90%. The parasite clearance time for groups E, F and G was (56.0 +/- 16.0), (53.3 +/- 4.6), and (56.0 +/- 8.0) h respectively, more rapid than that of Group H [(69.3 +/- 4.6) h]. There were 1, 3, 3, 2, 2, and 3 monkeys in groups A, B, D, E, F, and G respectively which were cured. No monkeys were cured in groups C, H and I. CONCLUSION: The combination of naphthoquine phosphate and artemisinine is superior to the single component and the optimum proportion in the combination is 1 : 2.5 in treating P. knowlesi infection in monkeys. PMID- 19288920 TI - [Staining improvement for Blastocystis hominis specimen]. AB - Schaudinn solution was used to fix the Blastocystis hominis specimen and an improved Harris hematoxylin staining was applied to stain it. The method shows clearer internal structure of the parasite, simpler and less time-consuming than the traditional iron hematoxylin solution. PMID- 19288921 TI - [Effect of maternal anti-Trichinella antibodies on intestinal worm burden in sucking mice]. AB - OBJECTIVE: To observe the transfer route of maternal anti-Trichinella antibodies and its effect on intestinal worm expulsion after the sucking mice was infected with T. spiralis. METHODS: Ninety-eight sucking mice (Kunming strain) were divided into 4 groups: mice born and nursed by infected mothers (group A), mice born of normal mothers and fostered by infected mothers (group B), mice born of infected mothers and fostered by normal mothers (group C), mice born of and nursed by normal mothers (group D). Blood was taken from tail veins of 4 groups of sucking mice when they were 14, 21 and 42 days old, respectively. Serum anti Trichinella antibody level was detected by ELISA using T. spiralis muscle larvae excretory-secretory (ES) antigens. Each sucking mouse were then orally challenged with 200 T. spiralis muscle larvae, and intestinal worm burden was observed 18 h after challenge infection. RESULTS: At 18 h after challenge infection, the mean intestinal worm burden of groups A, B, C and D of sucking mice aged 14 days was 5, 5, 19 and 18 larvae respectively. The worm burden of 4 groups of little mice aged 21 days was 18, 19, 75 and 73 larvae, respectively. Groups A and B of 14 and 21 day old mice fostered by infected mothers were shown to harbor obviously fewer worms than groups C and D of mice fostered by normal mothers (F14 = 10.056, F21 = 35.062, P < 0.01). Serum absorbance (A492) of groups A (0.177, 0.235) and B (0.183, 0.250) of 14 and 21 day old mice was significantly higher than groups C (0.108, 0.105) and D (0.067, 0.065) (F14 = 75.326, F21 = 60.867, P < 0.01). The intestinal worm burden in 4 groups of sucking mice aged 14 and 21 days showed significant negative correlation with their serum absorbance (r14 = -0.621, r21 = -0.756, P < 0.01). The intestinal worm burden in 4 groups of sucking mice aged 42 days was 55, 51, 46, and 60 larvae respectively, with no significant difference (F42 = 0.916, P > 0.05), their serum anti-Trichinella antibodies were negative. The intestinal worm burden in 4 groups of offspring mice aged 42 days showed no correlation with their serum absorbance (r42 = -0.291, P > 0.05). Anti Trichinella antibodies in sera of sucking mice were detected 6 hours after anti serum Ig to T. spiralis was intravenously injected into the lactating mothers. CONCLUSION: The maternal anti-Trichinella antibodies have been transferred from mother to filial mice mostly through milk, the antibodies can enhance the worm expulsion from intestine when the sucking mice aged 14-21 days was infected with T. spiralis larvae. no correlation with their serum absorbance (r42 = -0.291, P > 0.05). Anti-Trichinella antibodies in sera of sucking mice were detected 6 hours after anti-serum Ig to T. spiralis was intravenously injected into the lactating mothers. CONCLUSION: The maternal anti-Trichinella antibodies have been transferred from mother to filial mice mostly through milk, the antibodies can enhance the worm expulsion from intestine when the sucking mice aged 14-21 days was infected with T. spiralis larvae. PMID- 19288922 TI - [Epidemiological survey on paragonimiasis in Ningbo City, Zhejiang Province]. AB - Serum samples were collected from 2643 suspected cases of paragonimiasis in 2000 2007 from the outpatient departments of the city hospitals and surrounding areas, and the infection rate in the inhabitants, the first and second intermediate hosts, and animal reservoir hosts were investigated in the historical endemic areas. Serum samples were detected and 417 were found antibody positive (15.8%). Among residents in the historical endemic areas, the seropositive rate was 3.1% (46/1462), 2.8% (18/649) and 3.2% (26/813) in males and females respectively (CHI2 = 0.1833, P > 0.05). The infection rate in first intermediate host (snails), second intermediate host (crabs) and animal reservoir hosts was 0.05% (9/ 19,368), 31.1% (15,627/ 50,313) and 11.9% (52/438) respectively. Evidently, natural nidi for Paragonimus spp. still exist in Ningbo City. PMID- 19288923 TI - [In vitro culture of tachyzoites of Toxoplasma gondii RH strain in HeLa cells]. AB - OBJECTIVE: To establish a stable and efficient in vitro culture model for tachyzoites of Toxoplasma gondii RH strain. METHODS: Tachyzoites were inoculated into HeLa cells to establish an in vitro culture system. The proliferation of tachyzoites was observed under microscope by the method of Giemsa stain. At the same time, the longterm tachyzoites maintenance in HeLa cells was established, and the effect of different temperature and time on the yield and motility of tachyzoites were observed. RESULTS: The RH strain tachyzoites were cultured and maintained in HeLa cells. Most HeLa cells were destroyed 96 h after inoculation. In the long-term culture system, the proliferation of tachyzoites was stable and its virulence to mouse showed no decrease. Furthermore, tachyzoites in this system proliferated by 5-20 times and (1-5) x 10(7) tachyzoites were harvested. When cultured in HeLa cells at 37 degrees C for 72h then at 25 degrees C for another 120 h, the tachyzoites proliferated by more than 40 times with a motility rate of over 90%. However, rare HeLa cells left in the medium were found. CONCLUSION: Tachyzoites of T. gondii RH strain can be subcultured in HeLa cells for a long time, and high proliferation rate of tachyzoites can be obtained from this in vitro culture system. PMID- 19288924 TI - [Scan statistic method with moving window and its application in the temporal clustering detection of acute schistosomiasis]. AB - Acute schistosomiasis cases from 2001 to 2006 in Guichi District, Chizhou City, Anhui Province, were collected and analyzed by scan statistic method with moving window, central degree approach and circular distribution method in order to detect the temporal clustering of schistosomiasis. The analyzed data provided by the method showed that the temporal clustering of acute schistosomiasis from 2001 to 2003 in Guichi District became more concentrative, and then disperse, resulting in a final disappearance of the clustering in 2006 (LLR = 4.14, P > 0.05). The temporal clustering for all cases in the six years detected by scan statistic method was between August to September in 2002 (LLR = 18.5, P < 0.01), which was consistent with the real condition. Whereas the results from the central degree and the circular distribution methods could only provide limited information and were biased in the analysis in 2002, indicating that the scan statistic method with moving window can provide ample information with more robust and credible results. PMID- 19288925 TI - [In vitro observation on albendazole sulfoxide and its enantiomers against Echinococcus granulosus protoscolex]. AB - OBJECTIVE: To investigate in vitro anti-hydatid efficacy on Echinococcus granulosus protoscolex (EgPSC) by using albendazole sulfoxide (ASOX) and its two enantiomeric antipodes, L-ASOX and D-ASOX. METHODS: Eg protoscoleces were divided into eight groups and cultured in the DMEM culture media under two concentrations(50 microg/ml and 100 microg/ml) of ASOX, L-ASOX and D-ASOX respectively. The appropriate controls included (i) a culture containing an equal amount of DMSO and (ii) a culture medium alone. The mortality of EgPSC in each group was daily counted until 100% EgPSC death in some groups. RESULTS: Significant difference of EgPSC mortality was found among the three drugs with various concentrations compared to control group (P < 0.01), and a significant difference between L-ASOX group and D-ASOX group (P < 0.05). There were no statistical difference between ASOX group and D-ASOX group (P > 0.05), but between ASOX group and L-ASOX group (P < 0.05). On the 9th day of culture, the mortality of protoscoleces with the concentration of 50 microg/ml was 93.6%, 56.2% and 99.0% in ASOX, L-ASOX and D-ASOX groups respectively, and those under the concentration of 100 microg/ml were 100%, 74.5% and 100% respectively. The mortality was 19.1% and 22.5% respectively in the control and solvent groups. CONCLUSION: ASOX, L-ASOX and D-ASOX demonstrate significant effect of anti- Eg protoscolex in vitro. D-ASOX shows stronger effect than L-ASOX. PMID- 19288926 TI - [Anti-fecundity effect of soluble immature egg antigen of Schistosoma japonicum in mice]. AB - OBJECTIVE: To study anti-female fecundity effect in mice immunized with soluble immature egg antigen (SIEA) of Schistosoma japonicum, and observe possible mechanism. METHODS: Immature eggs were collected from the liver of a rabbit 34 days after being infected with 3000 cercariae of S. japonicum, and soluble antigen was prepared routinely. 20 ICR mice were divided randomly to 2 groups. 11 mice were immunized with SIEA (100 microg per mouse) and Freund adjuvant and the other 9 were injected with equal volume of sterile normal saline and Freund adjuvant as control. All mice received injection 5 times in 2 weeks interval. One week following the last vaccination, mice were challenged with 30 +/- 2 cercariae of S. japonicum Yunnan isolate. Feces were collected on day 45. All mice were sacrificed on day 46. The number of worms collected, number of eggs in feces, female uterus and liver, and the number of egg granulomas on the liver surface were compared between the 2 groups. The vitelline gland of female and testes of male worms were examined by transmission electron microscopy. RESULTS: There was no significant difference on the worm number between the immunized group and control (P > 0.05). The eggs per gram feces and liver, eggs in uterus per female, and egg granulomas on the liver surface were (56.68 +/- 24.78), (5 826 +/- 437), (49.94 +/- 12.53) and (10.04 +/- 1.13)/0.25 cm2, respectively in immunized group, while in control group these were (89.93 +/- 32.18), (10016 +/- 3541), (76.54 +/- 19.77) and (19.22 +/- 2.45)/0.25 cm2 respectively, all with significant difference (P < 0.05). Ultrastructure of the reproductive organs of paired adult worms showed that mature vitelline cells and lipid droplets in the cytoplasm of the cells decreased in the vitelline glands of immunized mice. More sustentacular cells and fewer spermatids were seen in testes of immunized mice. Vacuoles were seen in the cytoplasm of vitelline cells and sustentacular cells. CONCLUSION: The results indicated that SIEA may have an anti-fecundity effect possibly through inhibiting the maturation of germinal cells. PMID- 19288927 TI - [The structure and function of schistosome tegument and related proteomic study]. AB - Schistosome is covered by a living syncytium, called tegument, which plays important role in nutrient uptake and immune evasion. Recently, tegumental proteomic research identified lots of proteins, of which there may be potential targets for diagnosis, drugs and vaccines. PMID- 19288928 TI - [Field trial of malaria vaccine]. AB - Vaccine is expected to be a promising tool for malaria prevention and control. However, safe and effective malaria vaccine is not yet available for field use so far. They can be pre-erythrocytic stage vaccine, blood stage vaccine and transmission-blocking vaccine. This review summarizes the progress of the vaccine development in the recent field trials. PMID- 19288929 TI - [Limitations of the application of RNA interference in the study of animal parasitic nematodes]. AB - RNA interference is a powerful tool for investigating gene function which has been used extensively in model organisms such as Caenorhabditis elegans and has been adapted as a high-throughput screening method to identify genes. However, recent research indicates that the application of RNAi to animal parasitic nematodes has some problems. For example, the variability of RNAi to different parasitic nematodes or the same nematode in different stages is great. Reasons to explain why RNAi does not work well in animal parasitic nematodes include: (1) the efficacy of RNAi delivery methods in different nematodes is extremely variable; (2) RNAi mechanism has been applied to related nematodes and most of them are gene functional defect; (3) different lifestyles of nematodes may influence on the efficacy of RNAi. If we can find out the methods which can solve these questions, there are still application prospects for using RNAi in parasitic nematodes with improved RNAi effect. PMID- 19288930 TI - [An epidemiological survey on hydatid disease in Yushu Prefecture of Qinghai Province]. AB - Local inhabitants in 8 towns/townships of the counties Yushu, Zhiduo and Chengduo were examined with serology and ultrasound in 2006. Among 2 251 people tested by indirect hemagglutination (IHA), 207 showed anti-hydatid IgG positive (9.2%). Ultrasound examination found 106 cases out of 2581 people, with a morbidity of 4.1%. Females showed higher sero-positive rate (11.3%) and morbidity rate (5.0%) than males (6.6% and 3.0% respectively). The highest sero-positive rate was in the group of 40-49 years old (16.4%). The morbidity rate increased with age, with the highest rate (15.5%) in the group of 60 years old and above. Occupationally, those involved in semi-agriculture and semi-animal husbandry showed highest sero positive rate and morbidity rate, 21.5% and 11.4% respectively. The survey demonstrated that the prevalence of hydatid disease in human population is at a high level in Yushu Tibetan Autonomous Prefecture. PMID- 19288931 TI - [Cloning and eukaryotic expression of the gene encoding myosin from Brugia malayi]. AB - Total RNA was extracted from periodic microfilariae of Brugia malayi and its myosin partial gene (Bm-M55) was amplified by RT-PCR. The PCR product was cloned and then subcloned into pcDNA3.1 (+)vector. The recombinant eukaryotic plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and was transfected into COS-7 cells subsequently. The expressed protein was identified by SDS-PAGE. Bm-M55 mRNA was highly expressed in transfected COS-7 cells. The deduced amino acid sequence showed to be identical with that of Bm-M55, and the recombinant protein was about Mr 55000. PMID- 19288932 TI - [A case of peripheral T cell lymphoma complicated with visceral leishmaniasis]. PMID- 19288933 TI - Obesity in paediatric population of developing countries, a serious issue? PMID- 19288934 TI - Circadian pattern of onset of ischaemic and haemorrhagic strokes, and their relation to sleep/wake cycle. AB - OBJECTIVE: To determine whether there is a circadian pattern of onset of ischaemic and haemorrhagic strokes, and their relation to sleep/wake cycle. METHODS: A descriptive study with prospective data was conducted at the Combined Military Hospital Lahore from Jan 2004 to Dec 2007. Eight hundred patients above 26 years of age who had their first ever stroke were included in the study. Strokes were classified into cerebral infarction (CIF), intra-cerebral bleed (ICB) and subarachnoid haemorrhage (SAH). Diagnosis was confirmed either by CT or MRI scan of brain. RESULTS: Out of 800 patients, 80% were males and 20% were females. There were 438 (55%) cases of CIF, 329 (41%) of ICB and 33 (4%) of SAH. The age of the patients ranged from 26 to 84 years. Of all stroke cases, 592 (74%) occurred when the patients were awake and 208 (26%) occurred during sleep (p < 0.001). ICB cases showed significant variation with respect to wake/sleep cycle (p < 0.001). In CIF and SAH cases there was insignificant association with wake/sleep state of the patient, (p < 0.180 and 0.792 respectively). Of all strokes 22.5% occurred between 4 am - 8 am, followed by 20.7% between 4 pm - 8 pm, 20.1% between 8 am - 12 noon, 19.5% between 12 noon to 4 PM, 12.7% between 12 midnight and 4 am while 4.3% cases occurred between 8 pm and 12 midnight. The maximum number of CIF (28.5%) occurred between 4 am- 8 am, maximum ICB (29.8%) between 8 am to 12 noon and maximum SAH (30.3%) between 4 pm - 8 pm. The CIF and SAH cases showed smaller peaks between 4 pm to 8 pm and 8 am to 12 noon respectively. The lowest number of ICB cases (4.9%) were around mid night. Significant circadian variation was found in CIF and ICB patients (p < 0.001), however it was insignificant for SAH cases (p = 0.391). CONCLUSION: The findings of this study confirm the presence of circadian variation among cases of ischaemic stroke and intra cerebral bleed while no circadian variation was found in subarachnoid haemorrhage. CIF, ICB and SAH predominantly occur in early morning hours, late morning hours and in late afternoon to early evening respectively. Only intracerebral bleed was affected by wake/sleep state. PMID- 19288935 TI - Prevalence of human malaria infection in bordering areas of East Balochistan, adjoining with Punjab: Loralai and Musakhel. AB - OBJECTIVE: To study the prevalence of malarial infections in human population of districts Loralai and Musakhel areas of Pakistan. METHODS: Malarial parasites were identified in the blood slides of suspected patients of the disease from July, 2004 to June, 2006, and encompassed 7899 subjects. RESULTS: Out of 7899 suspected cases of malaria, 2275 (28.8%) were found to be positive for malarial parasite in blood smear slides. Out of positive cases, 1633 (71.7%) were identified as Plasmodium falciparum infection, 642 (28.2%) cases with P. vivax. However, seasonal variation was also noted with the highest (83.9%:287/342) infection of P. falciparum in September and lowest (65.3%: 34/52) in January in Loralai area whereas highest (76.9%:30/39) in October and lowest (3/9) in February in Musa Khel area. There was no case of Plasmodium malariae and P. ovale infection observed in the present study. These results are compared with those of other studies done in Pakistan. CONCLUSION: The high prevalence rate (71.7%:1633/2275) of P. falciparum poses a significant health hazard but 28.2% of P. vivax (642/2275) also may lead to serious complications like cerebral malaria. No association was found between types of infection and age groups. PMID- 19288936 TI - High prevalence of HIV infection among injection drug users (IDUs) in Hyderabad and Sukkur, Pakistan. AB - OBJECTIVE: To estimate size of injection drug users (IDU), determine their high risk behaviours and assess the prevalence of HIV. METHOD: As part of second generation surveillance (SGS), we investigated specific demographic and behavioural characteristics of IDUs in Hyderabad and Sukkur in 2005. It was a cross sectional study. The survey was preceded by geographic mapping to determine size estimation and to define sampling procedures prior to integrated behavioural and biological survey (IBBS). A sample size of at least 400 was calculated for each city. Besides calculating frequencies, chi square was used for comparing variables among HIV positive and negative IDUs like time elapsed as IDU, number of injections, sharing needles and self perception of acquiring HIV infection. RESULTS: A total of 800 (Hyderabad 398; Sukkur 402) questionnaires and DBS samples were collected. The estimated number of IDUs in both cities was 3,225 (Hyderabad 975 and Sukkur 2250 respectively). Average age of IDUs in Hyderabad was 36.5 years and 34.6 years in Sukkur. Sharing of injection equipment for last injection was reported by 34 (8.5%) in Hyderabad and 135 (33.6%) in Sukkur. In both cities behaviours such as injecting drugs for more than 10 years (p = 0.00) and injecting four or more times in a day (p = 0.11) were significantly associated with seropositivity of HIV infection. In Hyderabad the seroprevalence of HIV was 25.4% (101/398) and in Sukkur it was 19.2% (77/402). CONCLUSION: The burden of HIV among IDUs in Hyderabad and Sukkur is extremely high and can play a significant role in transmitting the infection to other vulnerable groups. PMID- 19288937 TI - Stenting in extracorporeal shockwave lithotripsy; may enhance the passage of the fragments! AB - OBJECTIVES: To prospectively evaluate the role of double J (DJ) stent in enhancing the passage of fragments in patients undergoing extracorporeal shockwave lithotripsy (SWL) for renal stones with diameters less than 2.5 cm. METHODS: Between November 2005 and January 2007, 38 patients with an average age of 47.05 years (range 16-73) were included and underwent SWL for renal stones. The inclusion criteria were radio-opaque renal stone not located in the lower pole, complete disintegeration of the stone, normal renal function, no metabolic abnormalities, no major renal abnormalities and no symptomatic urinary tract infection. The patients were randomized to either a stented (11 patients) or stentless (27 patients) group. The average stone diameters in stentless and stented groups were 1.54 cm and 1.77 cm, respectively (p > 0.05). Double J stent was removed when there was no further passage of the fragments for 6 weeks after stone disintegration. All patients were given non-steroidal anti-inflammatory drugs for one week after ESWL treatment. Stone passage and the data of DJ were determined with plain X-ray of the urinary tract (UTP). The severity of lower urinary tract symptoms, loin pain and the need for intravenous or intramuscular analgesics were recorded. RESULTS: The overall stone-free rate at 3 months was 92.1%. Two patients in the stented and one patient in the stentless group were partially free of stones. Steinstrasse were observed in two patients (5.3%); one patient in the stentless group and another one after the removal of DJ stent. Only one patient in the stented group had severe lower urinary tract symptoms which responded neither to oral nor to other forms of analgesics, and therefore DJ stent was removed. The remaining patients were in no need for analgesics other than the oral therapy. CONCLUSION: Placement of DJ stent for the purpose of improving free stone rate or enhancing the passage of the fragments during SWL is unnecessary in renal stone with diameters less than 2.5 cm. However, further prospective trials should be designed to define the criteria for stented SWL. PMID- 19288938 TI - The outcome of extracorporeal shockwave lithotripsy for renal pelvic stone with and without JJ stent--a comparative study. AB - OBJECTIVE: To compare the outcome of Extra corporeal shockwave lithotripsy for a renal pelvic stone with and without JJ stent. METHODS: A comparative cross sectional study was carried out at Sindh Institute of Urology and Transplantation from January 2007 to January 2008. Eighty patients with renal pelvic stone measuring 2cm +/- 2mm were selected for treatment with Extra Corporeal Shockwave Lithotripsy (ESWL). All of these patients were adults with normal renal function and had unilateral renal stones with negative urine cultures. Patients with renal failure and children were excluded. They were divided into two groups of 40 each. Group A patients underwent ESWL without a JJ stent and in Group B a JJ stent was placed before ESWL. SLX F2 electromagnetic ESWL machine was used to impart shock waves. 3000 shockwaves were given in a session. Both the groups were compared for renal colic, steinstrasse, fever, lower urinary tract symptoms (LUTS) emergency room visits and hospital admissions, stone clearance, number of ESWL sessions, auxilliary procedures, (percutaneous nephrostomy or ureterorenoscopy) and cost. RESULTS: Ureteric colic occurred in 13 (32.5%) patients in group A and in 3 (7.5%) patient in group B. Steinstrasse developed in 4 (10%) patients with out JJ stent and in 3 (7.5%) patients with JJ stent. Fever was encountered in 1 (2.5%) patient in group A and in 3 (7.5%) patient in group B. Mean emergency room visits were 2.1 per patient in group A and 0.7 per patient in group B. Stone clearance occurred in 33 (82.5%) patients in group A and 31 (77.5%) in group B. In group B lower urinary tract symptoms were found in 50% versus 20% in group A. Auxillary procedure was performed in one (2.5%) patient each in both groups. CONCLUSION: Pre ESWL JJ stenting for a 2 cm +/- 2 mm renal stone was not beneficial in terms of steinstrasse, fever, stone clearance and number of ESWL sessions. However ureteric colic was significantly less in the stented group. Lower urinary tract symptoms (LUTS) was also significantly high in the patients having a JJ stent. The cost of the treatment doubled in the stented group which is an important factor in our country. JJ stenting does not prove to be a cost effective procedure when compared to the reduction in complications. PMID- 19288939 TI - Elevated exhaled nitric oxide (NO) in asymptomatic asthmatics taking bronchodilators on demand with controlled body composition. AB - OBJECTIVE: Fractional Exhaled Nitric Oxide (FENO) is a recently introduced non invasive marker to measure inflammation and oxidative stress in the lung. This study aimed to measure FENO in Saudi asthmatic adult patients who had mild to moderate persistent asthma, on inhaled short-acting? 2 agonists and compared them to healthy individuals matched for body composition without any evidence of obstructive airway disease. METHODS: As per selection criteria 61 subjects were selected. 30 subjects were known asthmatic and 31 were healthy individuals matched for age, height, weight, BMI and body composition. Forced expiratory volume in 1 s (FEV1), FVC, FEV1/ FVC, PEF, FEF25, FEF50 and FEF75 were measured by standard methods. FENO measurements were performed according to the ATS (American Thoracic Society) recommendations. RESULTS: Ventillatory function parameters FEV1 (p = 0.0020), FVC (p = 0.0030), PEF (p = 0.0121), FEF25 (p = 0.0241), FEF50 (p = 0.0240) and FEF75 (p = 0.1824) were significantly lower in asthmatic subjects compared to matched healthy control group. FENO was significantly higher (82.51 +/- 39.26) in asthmatic subjects compared to control group (23.03 +/- 8.56) p < 0.0000) CONCLUSION: FENO levels are increased in patients with bronchial asthma with mild to moderate symptoms taking bronchodilators on demand only. It may be suggestive of the need for more accurate evaluation and early intervention with anti inflammatory drugs in a significant proportion of these patients. PMID- 19288940 TI - Childhood acute lymphoblastic leukaemia; epidemiology and clinicopathological features. AB - OBJECTIVE: To study epidemiology, clinical presentation and laboratory features of childhood Acute Lymphoblastic Leukaemia. METHOD: This retrospective review included all newly diagnosed children with acute lymphoblastic Leukaemia less than 15 years of age registered from April 1999 to December 2004 at oncology unit of National Institute of Child Health and Children Cancer Hospital, Karachi. The objective was to look for epidemiological data, the clinical features and laboratory findings at presentation and compare it with reported literature. RESULTS: Acute lymphoblastic Leukaemia constituted 32% (611 /1890) of all cancers in this study. Majority of patients hailed from Karachi (59%) and interior Sindh (27%) while rest from other parts of country. Patient's referral increased over the years, from 42 in 1999 to 127 in 2004. The age ranged between 3 months to 15 years with a median age of 6.5 years. Male to female ratio was 1.7:1 Family history of cancer was present in 5% of patients. Fever and pallor were the commonest presenting features. Anaemia (86%), lymphadenopathy (75%) hepatomegaly (67%) and splenomegly (58%) were common findings on physical examination. Initial high white cell count (> 50,000) was observed in 34% patients. Haemoglobin < 7gm/dl was seen in 54% and Platelet counts less than 20,000 was observed in 33% cases. CNS disease was present in 5% and HBsAg was positive in 14% patients at diagnosis. CONCLUSION: Acute Lymphoblastic Leukaemia accounts for one third of total registered cases. Age distribution in this series shows less prominemt early peak and more significant late peak and a median age of 6 years. Consangunity was found in 47% cases. The fraction with a WBC count above 50,000 mm3 (30%), a higher proportion with lymphadenopathy (75%) and haemoglobin less than 7 gm/dl (54%) suggest that Pakistani children have significantly higher burdens of Leukaemia cells at presentation. These may have prognostic implication resulting in poor outcome of Leukaemia in this part of the world. PMID- 19288941 TI - Success and failure of fast track extubation in cardiac surgery patients of tertiary care hospital: one year audit. AB - OBJECTIVE: To identify the causes of delayed extubation in patients planned for fast tract extubation during cardiac surgery. METHODS: A prospective observational study was conducted at cardiothoracic unit of Aga Khan University Hospital for the period of one year. All elective coronary artery bypass graft (CABG) surgery patients, between the ages of 35-75 years with LVEF > or = 40 percent were included. Patients with Intra aortic balloon pump, chronic renal failure, respiratory compromise and requiring high ionotropic support were excluded from this audit. A performa was designed and later filled by the primary investigator. Patient demographics and various reasons for delayed extubation were noted in this proforma. RESULTS: Total 614 patients underwent CABG surgery and 388 (63.19%) patients were planned for fast track extubation. A total of 196 (49.5%) patients could be extubated within six hours of arrival in the cardiac ICU. Common reasons for delayed extubation included deep sedation in 80 (46.5%), confusion 44 (25%), excessive bleeding in 20 (11.3%) and high inotropic support in 10 (5.68%). CONCLUSION: Major contributing factors for delayed extubation were identified by this audit. These factors need to be targeted accordingly by modifications in intra operative management. PMID- 19288942 TI - Open excision with secondary healing versus rhomboid excision with Limberg transposition flap in the management of sacrococcygeal pilonidal disease. AB - OBJECTIVE: To compare the outcome of Open excision and secondary healing with rhomboid excision and Limberg flap in the management of sacrococcygeal pilonidal sinus disease (SPD). METHODS: A prospective, analytical, comparative study using randomized controlled trial (RCT) was conducted at Liaquat University Hospital (Jamshoro) & Fatima Hospital-Baqai Medical University (Karachi), from June 2004 to July 2007. In total 49 patients, who either underwent open excision and secondary healing (group A: 25 patients) or rhomboid excision and Limberg flap (group B: 24 patients), were enrolled in the study. Duration of operation, postoperative pain, duration of hospital stay, postoperative complications, and time to recurrence were noted. The inclusion criteria were all patients with primary or recurrent SPD. The exclusion criteria were: cases with incomplete data and those lost to follow-up. RESULTS: Duration of operation was longer in group B patients (p = 0.004) but pain perception was markedly reduced in this group (p = 0.003). Total hospitalization period was shorter in patients in group B (p = 0.002) and so was the time for complete healing of the wound (p = 0.002). The recurrence rate was also significantly lower in patients who underwent Limberg rotation flap (p = 0.005). CONCLUSION: Limberg flap is advantageous over simple excision and secondary healing in the management of SPD. PMID- 19288943 TI - Fournier's gangrene--an experience in POF Hospital, Wah Cantt. AB - OBJECTIVE: To share our experience of treating patients with Fournier's Gangrene and their outcomes at POF Hospital, Wah Cantt. METHODS: A descriptive study was conducted at the Urology Department, POF Hospital, Wah Cantt from October 2006 to March 2007. A total of 22 patients, all men with ages ranging from 30-75 years, were included in the study. The diagnosis was established on the basis of history and clinical examination. The gangrenous patches with crepitus due to subcutaneous collection of seropurulent fluid was a diagnostic sign. RESULTS: A total of 22 patients were included in the study. All patients were men of ages ranging from 30 to 75 years. Of the 22 patients, 10 were diabetic, 02 had history of CVA and 10 had no co-morbids. In 10 patients, genitals and anterior abdominal wall was involved whereas 04 patients had genital and perineal involvement. In 05 patients only genitals were involved whereas 03 patients presented with scortal infection. All patients received broad spectrum antibiotics and underwent regular debridement. Secondary suturing was done in 13 patients, 07 patients needed skin grafting whereas orchidectomy was done in 02 patients. CONCLUSION: Fournier's gangrene is an uncommon but life threatening condition with high associated mortality and morbidity. Early diagnosis and treatment decrease the morbidity and mortality of this life threatening condition. PMID- 19288944 TI - Knowledge of medical students regarding diabetes mellitus at Ziauddin University, Karachi. AB - OBJECTIVE: To assess knowledge among medical students of Ziauddin University regarding Diabetes Mellitus. METHODS: A cross sectional survey of medical students from first year to final year at the medical college was conducted on a pretested self-administered questionnaire. Students were divided into preclinical and clinical years for analysis. RESULTS: A total of 366 students participated, with 145 (39.6%) from the preclinical group and 221 (60.4%) from the clinical group. Overall 89% students knew that diabetes is a metabolic disorder, 78% knew that pancreas is the site of insulin production and 77% students correctly identified main target organ of insulin. Regarding presentation, 67% said polyphagia, 50% said weight loss while only 18% claimed pruritus vulvae to be the presenting symptom. Majority from the clinical group (95%) while 86% of preclinical group knew about risk factors for diabetes (p = 0.003). Regarding prevention, 89% of clinical group knew that diabetes is preventable but 49% preclinical students did not have an idea of prevention (p < 0.001). A large number from clinical group (87%) said that there is some criteria of screening for diabetes while only 30% of preclinical group knew about screening (p < 0.001). Correct diagnostic criteria for diabetes according to WHO was identified by 55% clinical and only 6% of preclinical students (p < 0.001). Over two-third (78%) of clinical while only 50% of preclinical students said that diabetes should not always be treated with drugs (p < 0.001). CONCLUSION: The knowledge of students about diabetes was more in the clinical group as compared to the preclinical group, whereas, overall knowledge of the students was adequate. Medical students are the future physicians, therefore the medical curriculum should lay emphasis on educating students, patient based clinically oriented approaches for dealing with this epidemiologically important disease. PMID- 19288945 TI - Fine needle aspiration cytology of inflammatory breast lesions. AB - OBJECTIVE: To study the characteristics of inflammatory breast aspirate by Fine Needle Aspiration, performed by pathologist. METHODS: Inflammatory breast aspirates reported at two hospitals in Taif area (King Abdul Aziz Specialist Hospital and Al Hada Armed Forces Hospital, Taif, Kingdom of Saudi Arabia (KSA) from January 2000 till September 2007 were reviewed. The cytopathology reports, slides, clinical information, microbiology culture results, Tuberculosis; Polymerase Chain Reaction (TB; PCR) and histology were reviewed. The aspirates were identified as inflammatory based on presence of inflammatory cells and/or granulomas with occasional benign ductal epithelial cells. Cases of inflammatory carcinoma or duct ectasia were excluded. RESULTS: Forty-nine cases of inflammatory breast aspirates were included in the study. The aspirates were categorized into: Acute mastitis/Abscess 13 (26.5%), Granulomatous Mastitis 15 (30.6%), Fat Necrosis / Organized Haematoma 8 (16.4%) and Inflammation, not otherwise specified (NOS) 13 (26.5%), based on morphologic criteria combined with clinical impression. The Granulomatous Mastitis cases were subcategorized into Idiopathic and Infection related which included Brucellosis, Tuberculosis, and fungal mastitis. CONCLUSION: FNA of inflammatory breast lesions is a useful tool if performed by a pathologist and combined with further workup including microbiologic culture correlation. It helps in management and treatment of the patient and avoiding unnecessary surgery. PMID- 19288946 TI - Instituting an undergraduate core clerkship in radiology: initial experiences in Pakistan. AB - Utilization of radiology as an educational resource carries great potential. Accreditation bodies, physicians and medical students deem it important for a well rounded medical curriculum. Unfortunately this resource is yet to be developed and implemented to an optimal extent. We share the experiences from the first radiology core clerkship in Pakistan at the undergraduate level. An overview of clerkship objectives and structure is followed by discussion on lessons learnt during the initial three years of institution. Development of assessable objectives, integration of radiology with other specialties, and supervised andragogical learning tailored for undergraduate students are emphasized. PMID- 19288947 TI - Assessment of asthma control using the asthma control test at a tertiary care centre in Karachi, Pakistan. AB - The aim of our study was to assess asthma control among asthmatics at a tertiary care setting in Karachi using ACT questionnaire. The ACT questionnaire was filled by known asthmatics in January 2007. A total of 150 questionnaires were filled of which, 61 (40%) were males and 89 (60%) females. Mean ACT score was 17.71 +/- 4.41. Association between sex and asthma control was not statistically significant. Significant association was seen with asthma control and Ipratropium bromide inhaler. Asthma control among patients at a tertiary care centre is moderate. ACT can be used to follow patients in the hospital. PMID- 19288948 TI - Dengue fever with hepatitis E and hepatitis A infection. AB - Infection with dengue viruses produces a spectrum of clinical illness ranging from a nonspecific viral syndrome to severe and fatal haemorrhagic disease. Important risk factors include the strain and serotype of the infecting virus, as well as the age, immune status, and genetic predisposition of the patient. The teaching point in this case study was Dengue fever which occurred concomitantly with Hepatitis A and Hepatitis E virus infection. PMID- 19288949 TI - Tramadol overdose induced seizure, dramatic rise of CPK and acute renal failure. AB - Tramadol, an analgesic with a low affinity to opioid receptors, inhibits the reuptake of norepinephrin and serotonin. It is also abused by opioid addicts. Tramadol overdose can induce CNS and respiratory depression, tachycardia, and seizures. In this report, a 19 years male was admitted due to suicidal attempt of ingestion of 4000 mg of Tramadol. He experienced frequent seizures, confusion, myosis, and dramaticrise of CPK, LDH and Creatinine. Improvement was had in the following days by administering fluids, NaHCO3 and chlordiasopoxide and routine management. He was discharged with no further sequelae. PMID- 19288950 TI - Primary bone lymphoma: a series from a cancer institute in Pakistan. AB - Primary bone lymphoma (PBL) is an uncommon tumour. Numerous studies have been reported from Western countries, but none from Southeast Asia. We reviewed a series of seven consecutive patients diagnosed and treated with PBL at our hospital between March 2002 and January 2007. All patients underwent chemotherapy with half receiving radiotherapy as their initial treatment. Six (84%) patients were male and 1 (16%) female with a median age of 33 (range: 23-85). All had diffuse large B-cell lymphoma (DLBCL) of bone except one (85 - M) who had chest wall cutaneous T-cell lymphoma with iliac blade involvement. The femur was the most frequently involved site (43%). Except for three patients that involved the lymph nodes, all patients had disease limited to bone. The 5-year overall survival rate was 43%. Although the number of patients was small, the data presented here revealed several characteristics of PBL. PMID- 19288951 TI - Persistent appendiceal faecal fistula following a complicated open appendicectomy. AB - Appendiceal faecal fistula is recognized serious complication of appendicectomy, which is one of the commonest surgical procedures performed in modern day surgical practice. We report a unique case of appendiceal faecal fistula which persisted for nine years in a teenage girl. PMID- 19288952 TI - Ileal ureteral replacement in a man with Studer pouch. AB - Use of a bowel segment for ureteral replacement is a reliable and a feasible procedure with satisfactory results. We present a patient with a complete left ureteral necrosis due to infection; with an abscess formation in the retroperitoneum after a radical cystoprostatectomy and Studer pouch operation. PMID- 19288953 TI - 'Homosexuality: are we adequately equipped to address this issue in clinical practice'? PMID- 19288954 TI - "Alcohol use in mouthwash and possible oral health concerns". AB - OBJECTIVE: To establish the presence and quantify Ethanol in commercially available mouthwashes. METHODS: Samples from twelve commercially available mouthwashes were tested for the presence of Ethanol followed by the estimation of percentage of Ethanol in five brands in Pakistan Council of Scientific and Industrial Research (P.C.S.I.R) and Husein Ebrahim Jamal (H.E.J.) labs, Karachi. RESULTS: Ten out of twelve brands of mouthwashes were found to be Ethanol positive. CONCLUSION: Alcohol (Ethanol) in the mouthwashes does not contribute to any therapeutic action. It is alarming to find the presence of alcohol in the mouthwashes which claim to contain no alcohol. PMID- 19288955 TI - Communication & inter-personal skills? But I'm a radiologist... PMID- 19288956 TI - Treatment of achalasia: lets put surgery in its right context. PMID- 19288957 TI - Assessment and simulation of the implementation of brucellosis control programme in an endemic area of the Middle East. AB - Brucellosis is an important zoonosis in Middle Eastern countries. In this study we assessed the extent of the application of planned official brucellosis control programmes in Kafr El Sheikh governorate, Egypt and we used a stochastic simulation model to assess the probable impact of changes to the official control strategy on the dynamics of small-ruminant brucellosis. Our results show that <7% of female livestock were tested in any given year in the study area and that quarantine was not consistently applied to infected herds. Simulation results revealed the inability of the applied control measures to reduce the prevalence of small-ruminant brucellosis. Given our assumptions, the intensity with which infected animals are removed under the actual levels of implementation of test and-slaughter programmes would permit brucellosis to remain endemic at a level >8% of the sheep and goat population. PMID- 19288958 TI - Survey and case-control study during epidemics of Puumala virus infection. AB - In Baden-Wuerttemberg, a federal state in south-west Germany, a large outbreak of 1089 laboratory-confirmed human Puumala virus (PUUV) infections occurred in 2007. We conducted a survey to describe the disease burden and a case-control study to identify risk factors for acquiring PUUV. Case-patients were interviewed about clinical outcome and both case-patients and randomly recruited controls were interviewed about exposure. We calculated matched odds ratios (mOR) using a conditional logistic regression model. Multivariable analysis of 191 matched case control pairs showed that case-patients were more likely than controls to have seen small rodents/their droppings (mOR 1.9, 95% CI 1.2-3.0), cleaned utility rooms (mOR 1.8, 95% CI 1.0-3.4) and visited forest shelters (mOR 3.9, 95% CI 1.1 14.3). Two thirds of case-patients required hospitalization. During PUUV epidemics rodent control measures and use of protective equipment should be considered in utility rooms and shelters. PMID- 19288959 TI - New reports of Australian cutaneous leishmaniasis in Northern Australian macropods. AB - Cutaneous leishmaniasis caused by various species of Leishmania is a significant zoonotic disease in many parts of the world. We describe the first cases of Australian cutaneous leishmaniasis in eight northern wallaroos, one black wallaroo and two agile wallabies from the Northern Territory of Australia. Diagnosis was made through a combination of gross appearance of lesions, cytology, histology, direct culture, serology and a species-specific real-time PCR. The causative organism was found to be the same unique species of Leishmania previously identified in red kangaroos. These clinical findings provide further evidence for the continuous transmission of the Australian Leishmania species and its presence highlights the importance of continued monitoring and research into the life-cycle of this parasite. PMID- 19288960 TI - The effect of vaccination on undetected persistence of foot-and-mouth disease virus in cattle herds and sheep flocks. AB - The importance of carrier animals (those in whom virus persists after recovery from disease or acute infection) and their potential role in the spread of disease remain open questions within foot-and-mouth disease epidemiology. Using simple probabilistic models we attempt to quantify the effect of emergency vaccination--and especially the time of application--on the likely number of such animals, using data from challenge experiments on both cattle and sheep to determine the probability of persistence in diseased and subclinically infected animals. We show that the number of persistently infected animals in a group is predominantly determined by the number of animals initially infected on premises- the high variability of which ultimately limits the accuracy of any predictions of carrier numbers based upon transmission models. Furthermore, results suggest that, within a cattle herd, carrier numbers may be increased if challenge occurs shortly after vaccination. We show that the quality of inspection is the principal factor influencing whether or not carrier herds occur and that, by reducing clinical signs, the application of vaccination in regularly checked stock also results in an increase in undetected persistently infected animals. Where clinical detection would be poor regardless of the use of vaccination (i.e. particularly in sheep), vaccination will result in a reduction in the probability of a group containing undetected carriers: otherwise there is a benefit only if vaccination is applied sufficiently far in advance of any challenge. The implications of the results for serosurveillance are discussed, including the requisite test sensitivity and practices for successful implementation. PMID- 19288961 TI - The continuous spread of West Nile virus (WNV): seroprevalence in asymptomatic horses. AB - West Nile virus (WNV) was probably introduced in southern and northern Mexico from the USA in two independent events. Since then, WNV activity has been reported in several Mexican states bordering the USA and the Gulf of Mexico, but disease manifestations seen there in humans and equids are quite different to those observed in the USA. We have analysed WNV seroprevalence in asymptomatic, unvaccinated equids from two Mexican states where no data had been previously recorded. WNV IgG antibodies were detected in 31.6% (91/288) of equine sera from Chiapas and Puebla states (53.3% and 8.0%, respectively). Analysis by plaque reduction neutralization test (PRNT) showed good specificity (99.4%) and sensitivity (84.9%) with the ELISA results. Further analyses to detect antibodies against three different flaviviruses (WNV, St Louis encephalitis virus, Ilheus virus) by haemagglutination inhibition (HI) tests on a subset of 138 samples showed that 53% of the 83 HI-positive samples showed specific reaction to WNV. These data suggest continuous expansion of WNV through Mexico. PMID- 19288962 TI - Bridging the gap: an innovative dementia learning program for healthcare assistants in hospital wards using facilitator-led discussions. AB - Nursing a person with dementia in a ward setting can be stressful and a challenge for staff and patients alike. Healthcare assistants are identified as requiring a specific training program. They form part of the front-line workforce and yet have the least access to training but often most contact with patients. The program in this study focused on person-centered care and used six self-study workbooks. Experienced registered nurses are trained to be facilitators of 12 group discussions in the ward setting. The training program viewed the facilitator as playing a key role in empowering the healthcare assistant but also in promoting reflective practice. The outcomes to date have been positive and showed a development in confidence and competence of the healthcare assistants involved. PMID- 19288963 TI - Educating primary care physicians in the management of Alzheimer's disease: using practice guidelines to set quality benchmarks. AB - This paper presents a strategy for training primary care physicians in the identification, diagnosis and management of Alzheimer's disease and related disorders. The strategy uses evidence-based practice guidelines to establish quality benchmarks and then provides training and other interventions to improve the quality of care received by these patients. The three projects described in this paper assumed that training of primary care physicians alone would not be sufficient to achieve the quality benchmarks derived from guidelines. The projects used creative training strategies supplemented by provider "tool kits", provider checklists, educational detailing, and endorsement from organizational leadership to reinforce what the primary care providers learned in educational sessions. Each project also implemented a system of dementia care management to "wrap around" traditional primary care to ensure that quality benchmarks would be achieved. Outcomes of two completed studies support the premise that it is possible to improve quality of dementia care through physician education that occurs in association with a coordinated system of dementia care management and in collaboration with community agencies to access guideline-recommended social services. PMID- 19288964 TI - Translating dementia research into practice. AB - There is a gap between the knowledge obtained in dementia research and the practical use of it. This review examines the concept of knowledge translation (KT) and the process of translating research into practice in the field of dementia. KT in dementia practice involves key players including researchers, educators, clinicians, policy-makers, the general public and consumers who act within a multidimensional network, disseminating findings widely to effect changes in community awareness, clinical practice and health policy. Challenges include the volume of dementia research; difficulties in determining who is responsible for KT; the problems of premature KT; the lack of resources to perform KT; the paucity of research about effective KT strategies; and the characteristics of the dementia care workforce. Key features of effective KT include a simple compelling message; use of interpersonal contact and roles; a practical framework with an emphasis on "know-how"; and the provision of resources and support. More effective dementia KT will require a commitment by key players to engage in the process, a better understanding of effective KT strategies, adequate resources, and judicious selection of appropriate evidence based information. PMID- 19288965 TI - Snapshot: Alzheimer's Australia pilot of an innovative approach to consumer education and training. AB - BACKGROUND: Enabling the families and carers of people with dementia to access information, support and opportunities for education and training is critical to ensuring that both the carer and person with dementia maintain their quality of life. The Dementia Caring Pilot Project trialed an innovative approach to providing this support through offering tailored activities aimed at enhancing and developing a range of skills, both dementia-specific and life oriented. METHOD: Alzheimer's Australia worked closely with Commonwealth Carer Respite Centres (CCRCs) across Australia to identify potential participants, and then deliver skills enhancement activities that were suited to their individual needs. RESULTS: Nationally, 47 CCRCs participated in the Project and 324 skills enhancement activities were provided to over 1000 participants. CONCLUSIONS: The Project was successful in giving the families and carers of people with dementia the opportunity to express the need for the capabilities they wished to enhance, and to have the opportunity of a flexible service response. PMID- 19288966 TI - Large-scale training in the essentials of dementia care in Australia: Dementia Care Skills for Aged Care Workers project. AB - Dementia has been identified as a national health priority in Australia. National programs in the areas of research, education and training have been established. The Dementia Care Skills for Aged Care Workers program is a three-year project that commenced in 2006. It has the goal of providing training in the essentials of dementia care to 17,000 staff of aged care services across Australia. Successful completion of the training results in the award of a nationally recognized qualification. Although the delivery of the training has been difficult in some areas--because of the long distances to be covered by trainers and trainees, a wide range of cultural backgrounds, and difficulties in finding staff to cover for people attending the training--the seven training organizations providing this training are on target to meet the goal. The project is being evaluated independently. The anecdotal reports available to date strongly suggest that the training is being well received and is making a difference to practice.A surprisingly large proportion of attendees (9%) have been registered nurses, which demonstrates the need among this group of staff for training in the care of people with dementia. PMID- 19288967 TI - International perspectives on dementia education, training and knowledge transfer. PMID- 19288968 TI - Alzheimer's Association Quality Care Campaign and professional training initiatives: improving hands-on care for people with dementia in the U.S.A. AB - In the U.S.A., direct care workers and licensed practical nurses are the professionals who provide the most hands-on care to people with dementia in nursing homes and residential care facilities--yet they do not receive adequate training in dementia care. Dementia care training needs to be universal with all disciplines at all levels of care. Even though there is variability on recommended hours and content, most studies emphasize the importance of dementia care training as a distinct component of required training for any professional or paraprofessional working in long-term care. In 2005, the Alzheimer's Association launched its Quality Care Campaign to improve dementia care through state and federal advocacy; consumer education and empowerment; and staff training. This paper describes the effectiveness of Alzheimer's Association training as measured by knowledge gained and providers' intention to change their behavior immediately after attending the training.Overall, findings indicated that the participants responded positively to evidence-based training in dementia care that emphasized the importance of (i) leadership, (ii) team communication and collaboration, (iii) support and empowerment of direct care staff, (iv) awareness and practice of specific dementia care issues, (v) resident and family involvement in care, and (vi) professional self-care. PMID- 19288969 TI - Realizing a palliative approach in dementia care: strategies to facilitate aged care staff engagement in evidence-based practice. AB - BACKGROUND: There is growing evidence that a palliative approach to care provision for people with dementia in residential aged care facilities improves their quality of life and provides support for family members. Despite the development of Guidelines for a Palliative Approach in Residential Aged Care (hereafter the Guidelines), there is limited evidence that these have been adopted. To date, little research has been undertaken to explore processes which could assist aged care staff to develop their practice consistent with the intent of the Guidelines. METHODS: This project utilized an action research method, through which staff members from a residential dementia special care unit (SCU) undertook an investigation into their practice to explore how they could develop strategies to support a palliative approach to care provision. A key focus was related to addressing the information needs of family members of residents on the SCU. RESULTS: Aged care staff involved in this project had little understanding of available evidence that could assist them to better support family members, including the existence of the Guidelines. Through their engagement in successive action research cycles, these staff accessed evidence-based resources and developed strategies to address the information needs of family members. CONCLUSIONS: When provided with an opportunity to reflect on and critique their practice, aged care staff were better positioned to engage with evidence concerning a palliative approach and to execute change in their practice to improve care provision for family members. PMID- 19288970 TI - The effect of continuing professional education on health care outcomes: lessons for dementia care. AB - BACKGROUND: This review presents an overview of the published literature on the effectiveness of continuing professional education (CPE), which includes continuing medical education (CME) of different health care professionals in healthcare settings, for improving patient management and patient outcomes. This review summarizes key articles published on the subject, including those relating to dementia care. METHODS: A literature search was carried out using the National Library of Medicine's PubMed database, Cochrane database and Eric databases. RESULTS: Studies on CPE generally provide conflicting evidence on their effectiveness in bringing about a change in professional practices and healthcare outcomes. However interactive, multifaceted interventions, and interventions with repeated inputs appear more effective in bringing about positive changes than traditional non-interactive techniques. There are relatively few studies specifically concerning CPE and dementia care. CONCLUSION: This review shows that CPE in dementia care needs to be targeted carefully. Much can be learnt from examining education approaches in the wider professional and medical education literature. PMID- 19288971 TI - Minimizing attrition bias: a longitudinal study of depressive symptoms in an elderly cohort. AB - BACKGROUND: Attrition from mortality is common in longitudinal studies of the elderly. Ignoring the resulting non-response or missing data can bias study results. METHODS: 1260 elderly participants underwent biennial follow-up assessments over 10 years. Many missed one or more assessments over this period. We compared three statistical models to evaluate the impact of missing data on an analysis of depressive symptoms over time. The first analytic model (generalized mixed model) treated non-response as data missing at random. The other two models used shared parameter methods; each had different specifications for dropout but both jointly modeled both outcome and dropout through a common random effect. RESULTS: The presence of depressive symptoms was associated with being female, having less education, functional impairment, using more prescription drugs, and taking antidepressant drugs. In all three models, the same variables were significantly associated with depression and in the same direction. However, the strength of the associations differed widely between the generalized mixed model and the shared parameter models. Although the two shared parameter models had different assumptions about the dropout process, they yielded similar estimates for the outcome. One model fitted the data better, and the other was computationally faster. CONCLUSIONS: Dropout does not occur randomly in longitudinal studies of the elderly. Thus, simply ignoring it can yield biased results. Shared parameter models are a powerful, flexible, and easily implemented tool for analyzing longitudinal data while minimizing bias due to nonrandom attrition. PMID- 19288972 TI - Leadership: a central tenet for postgraduate dementia services curricula development in Australia. AB - BACKGROUND: In the next decades of the twenty-first century, the global aging of populations will challenge every nation's ability to provide leadership by qualified health professionals to reshape and improve health care delivery systems. The challenge for educators is to design and deliver courses that will give students the knowledge and skills they need to fill that leadership role confidently in dementia care services. This paper explores the ways in which a curriculum can develop graduates who are ready to become leaders in shaping their industry. METHOD: The Master of Health Science-Aged Services (MHSAS) program at Victoria University, Melbourne, Australia is applied as a case study to describe the process by which the concept of leadership is applied as the key driver in curriculum development, teaching practices and learning outcomes. RESULTS: Evaluation instruments employed in a variety of purposes including teaching, curriculum planning and unit appraisal are discussed. Challenges for the future are proposed including the need for postgraduate programs in dementia to seek stronger national and international benchmarks and associations with other educational institutions to promote leadership and a vision of what is possible and desirable in dementia care provision. CONCLUSIONS: In the twenty-first century, effective service provision in the aged health care sector will require postgraduate curricula that equip students for dementia care leadership. The MHSAS program provides an established template for such curricula. PMID- 19288973 TI - Effects of chronic and acute consumption of fruit- and vegetable-puree-based drinks on vasodilation, risk factors for CVD and the response as a result of the eNOS G298T polymorphism. AB - The average UK adult consumes less than three portions of fruit and vegetables daily, despite evidence to suggest that consuming five portions daily could help prevent chronic diseases. It is recommended that fruit juice should only count as one of these portions, as juicing removes fibre and releases sugars. However, fruit juices contain beneficial compounds such as vitamin C and flavonoids and could be a useful source of dietary phytochemicals. Two randomised controlled cross-over intervention studies investigating the effects of chronic and acute consumption of commercially-available fruit- and vegetable-puree-based drinks (FVPD) on bioavailability, antioxidant status and CVD risk factors are described. Blood and urine samples were collected during both studies and vascular tone was measured using laser Doppler imaging. In the chronic intervention study FVPD consumption was found to significantly increase dietary carotenoids (P=0.001) and vitamin C (P=0.003). Plasma carotenoids were increased (P=0.001), but the increase in plasma vitamin C was not significant. There were no significant effects on oxidative stress, antioxidant status and other CVD risk factors. In the acute intervention study FVPD were found to increase total plasma nitrate and nitrite (P=0.001) and plasma vitamin C (P=0.002). There was no effect on plasma lipids or uric acid, but there was a lower glucose and insulin peak concentration after consumption of the FVPD compared with the sugar-matched control. There was a trend towards increased vasodilation following both chronic and acute FVPD consumption. All volunteers were retrospectively genotyped for the eNOS G298T polymorphism and the effect of genotype on the measurements is discussed. Overall, there was a non-significant trend towards increased endothelium dependent vasodilation following both acute and chronic FVPD consumption. However, there was a significant time x treatment effect (P<0.05) of acute FVPD consumption in individuals with the GG variant of the eNOS gene. PMID- 19288974 TI - Neurotoxic effects induced by gammahydroxybutyric acid (GHB) in male rats. AB - Gammahydroxybutyric acid (GHB) is an endogenous constituent of the central nervous system that has acquired great social relevance for its use as a recreational 'club drug'. GHB, popularly known as 'liquid ecstasy', is addictive when used continuously. Although the symptoms associated with acute intoxication are well known, the effects of prolonged use remain uncertain. We examined in male rats the effect of repeated administration of GHB (10 and 100 mg/kg) on various parameters: neurological damage, working memory and spatial memory, using neurological tests, the Morris water maze and the hole-board test. The results showed that repeated administration of GHB, especially at doses of 10 mg/kg, causes neurological damage, affecting the 'grasping' reflex, as well as alteration in spatial and working memories. Stereological quantification showed that this drug produces a drastic neuronal loss in the CA1 hippocampal region and in the prefrontal cortex, two areas clearly involved in cognitive and neurological functions. No effects were noted after quantification in the periaqueductal grey matter (PAG), a region lacking GHB receptors. Moreover, NCS 382, a putative antagonist of GHB receptor, prevented both neurological damage and working- memory impairment induced by GHB. This suggests that the effects of administration of this compound may be mediated, at least partly, by specific receptors in the nervous system. The results show for the first time that the repeated administration of GHB, especially at very low doses, produces neurotoxic effects. This is very relevant because its abuse, especially by young persons, could produce considerable neurological alterations after prolonged abuse. PMID- 19288975 TI - Riluzole for relapse prevention following intravenous ketamine in treatment resistant depression: a pilot randomized, placebo-controlled continuation trial. AB - The N-methyl-D-aspartate (NMDA) glutamate receptor antagonist ketamine may have rapid, albeit transient, antidepressant properties. This study in patients with treatment-resistant major depression (TRD) aimed to (1) replicate the acute efficacy of single-dose intravenous (i.v.) ketamine; (2) test the efficacy of the glutamate-modulating agent riluzole in preventing post-ketamine relapse; and (3) examine whether pretreatment with lamotrigine would attenuate ketamine's psychotomimetic effects and enhance its antidepressant activity. Twenty-six medication-free patients received open-label i.v. ketamine (0.5 mg/kg over 40 min). Two hours prior to infusion, patients were randomized to lamotrigine (300 mg) or placebo. Seventeen patients (65%) met response criterion (50% reduction from baseline on the Montgomery-Asberg Depression Rating Scale) 24 h following ketamine. Lamotrigine failed to attenuate the mild, transient side-effects associated with ketamine and did not enhance its antidepressant effects. Fourteen patients (54%) met response criterion 72 h following ketamine and proceeded to participate in a 32-d, randomized, double-blind, placebo-controlled, flexible dose continuation trial of riluzole (100-200 mg/d). The main outcome measure was time-to-relapse. An interim analysis found no significant differences in time-to relapse between riluzole and placebo groups [log-rank chi(2) = 0.17, d.f. = 1, p = 0.68], with 80% of patients relapsing on riluzole vs. 50% on placebo. The trial was thus stopped for futility. This pilot study showed that a sub-anaesthetic dose of i.v. ketamine is well-tolerated in TRD, and may have rapid and sustained antidepressant properties. Riluzole did not prevent relapse in the first month following ketamine. Further investigation of relapse prevention strategies post ketamine is necessary. PMID- 19288976 TI - The impact of social influence on adolescent intention to smoke: combining types and referents of influence. AB - OBJECTIVES: Theory and research suggest that the intention to smoke is the main determinant of smoking initiation and emphasizes the role of cognitive and social factors on the prediction of the intention to smoke. However, extended models such as the I-Change and results from published studies reveal inconsistencies regarding the impact of social influence on the intention to smoke. Possible explanations for this may be the definition and measurement of the constructs that have been used. DESIGN AND METHODS: The current study was designed with two main goals: (i) to test a measurement model for social influence, combining different types of social influence (subjective norms, perceived behaviour, and direct pressure) with various referents of influence (parents, siblings, peers, and teachers); (ii) to investigate the impact of social influence on adolescent intention to smoke, controlling for smoking behaviour. LISREL was used to test these models. The sample includes 3,064 Portuguese adolescents, with a mean age of 13.5 years, at the beginning of the seventh school grade. RESULTS: The hypothesized measurement model of social influence was supported by results and explained 29% of the variance of the intention to smoke. A more extended model, including attitude and self-efficacy, explained 55% of the variance of the intention to smoke. Perceived behaviour of peers, parental norms, and perceived behaviour of parents were the social influence factors with impact on adolescent intention to smoke. CONCLUSIONS: Results suggest that different referents exert their influence through distinct types of social influence and recommend further work on the definition and measurement of social influence. PMID- 19288977 TI - The comprehension skills of children learning English as an additional language. AB - BACKGROUND: Data from national test results suggests that children who are learning English as an additional language (EAL) experience relatively lower levels of educational attainment in comparison to their monolingual, English speaking peers. AIMS: The relative underachievement of children who are learning EAL demands that the literacy needs of this group are identified. To this end, this study aimed to explore the reading- and comprehension-related skills of a group of EAL learners. SAMPLE: Data are reported from 92 Year 3 pupils, of whom 46 children are learning EAL. METHOD: Children completed standardized measures of reading accuracy and comprehension, listening comprehension, and receptive and expressive vocabulary. RESULTS: Results indicate that many EAL learners experience difficulties in understanding written and spoken text. These comprehension difficulties are not related to decoding problems but are related to significantly lower levels of vocabulary knowledge experienced by this group. CONCLUSIONS: Many EAL learners experience significantly lower levels of English vocabulary knowledge which has a significant impact on their ability to understand written and spoken text. Greater emphasis on language development is therefore needed in the school curriculum to attempt to address the limited language skills of children learning EAL. PMID- 19288978 TI - Adult attachment in bipolar 1 disorder. AB - OBJECTIVES: To determine how security of adult attachment style is related to the mania, major depression and euthymic mood states in bipolar 1 (BP1) disorder. DESIGN: An observational cross-sectional study. METHOD: One hundred and seven BP1 patients (34 in a manic type episode, 30 in major depressive episode, and 43 in remission) and 41 healthy controls similar in age, gender, reading age, and education were recruited. The groups were compared on self-reported mean and preferred attachment style controlling for psychiatric comorbidity. RESULTS: Preferred attachment style was insecure in 84 (78%) BP1 patients but only 13 (32%) healthy controls (chi(2)=34.3, df=3, and p<.001). Healthy controls reported higher secure attachment, lower anxious, and lower preoccupied attachment scores than all groups of patients with bipolar disorder, although the scores for secure attachment in mania and preoccupied attachment in euthymic patients were not significantly different from healthy controls. Overall, within the bipolar groups, anxious attachment style varied little with mood but mania was associated with higher secure and preoccupied attachment style, and depression with higher preoccupied and lower dismissing attachment style scores. CONCLUSIONS: Insecure attachment is found in most patients with BP1 disorder. Attachment style is affected by mood episodes so it should be assessed when a patient with bipolar disorder is in remission with minimal residual depressive or manic symptoms. PMID- 19288979 TI - One size does not fit all: motivational predictors of contextual benefits of therapy. AB - BACKGROUND: A common sense view in psychotherapy holds that there are individual differences in response to different treatments. However, despite considerable effort, no clear rationale exists to guide the selection of therapy for individual patients. OBJECTIVES: Rather than focusing on specific components of therapy as a basis to predict interactions between patients and treatments, we draw on motivational concordance theory and a contextual model of psychotherapy to test whether, in an analogue study, perceived effectiveness of different therapy vignettes is related to human values. METHOD: Two samples (undergraduates and members of the public) read short vignettes, informed by six therapies for depression (cognitive behaviour therapy, client-centred therapy, antidepressant medication, existential psychotherapy, psychodynamic psychotherapy, and a complementary medicine: Bach flower essences), provided ratings of perceived effectiveness, and completed questionnaire measures of dispositional motivations (Schwartz Values Scale and the Spiritual Connection Questionnaire-14). Responses were analysed using multidimensional scaling (MDS). RESULTS: In both samples, expectancy for the cognitive behavioural-informed vignette was related to Self direction and Stimulation, and was opposed to Tradition, Conformity and Security. Expectancies for the Drug vignette were associated with Power and Security. CONCLUSION: People perceive therapies as more effective if the therapy is congruent with their values. These preliminary data suggest motivational concordance is a useful framework for predicting client-treatment interactions. PMID- 19288980 TI - A new protocol for specific oral tolerance induction in children with IgE mediated cow's milk allergy. AB - IgE-mediated cow's milk allergy (CMA) is a heavy burden for patients, particularly for children and their families. Allergen avoidance represents the only therapeutic option, but oral desensitization protocols have been suggested. Because of the long duration and complexity of these protocols we examined the feasibility of an oral tolerance induction protocol using a weekly up-dosing schedule. Children with IgE-mediated food allergy to milk, confirmed by a double blind placebo-controlled food challenge, were recruited. Six of them were randomized to double-blind desensitization with milk or soy formula as placebo. Seven patients underwent the protocol in open fashion. The desensitization schedule started with one drop of whole CM diluted 1:25 every week. The dose was doubled weekly until the 18th week to achieve an intake of 200 mL in approximately 4 months. Of the 13 children enrolled, 10 children received CM and 3 control children received soy formula. Full tolerance (200 mL of milk) was achieved in 7 children; in 2 children this therapeutic approach failed, because severe reactions occurred during the procedure. One patient achieved a partial tolerance (64 mL of milk). The three control children receiving placebo still showed a positive food challenge at the end of the study. A weekly up-dosing oral tolerance induction could be a viable alternative to traditional protocols for children with IgE-mediated CMA. PMID- 19288981 TI - Public-private partnerships and public health practice in the 21st century: looking back at the experience of the Steps Program. PMID- 19288982 TI - Public health's inconvenient truth: the need to create partnerships with the business sector. PMID- 19288983 TI - Communication inequalities, social determinants, and intermittent smoking in the 2003 Health Information National Trends Survey. AB - INTRODUCTION: Intermittent smokers account for a large proportion of all smokers, and this trend is increasing. Social and communication inequalities may account for disparities in intermittent smoking status. METHODS: Data for this study came from 2,641 ever-smokers from a 2003 nationally representative cross-sectional survey. Independent variables of interest included race/ethnicity, sex, household income, education, health media attention, and cancer-related beliefs. The outcome of interest was smoking status categorized as daily smoker, intermittent smoker, or former smoker. Analyses used 2 sets of multivariable logistic regressions to investigate the associations of covariates with intermittent smokers compared with former smokers and with daily smokers. RESULTS: People with high education and high income, Spanish-speaking Hispanics, and women were the most likely to be intermittent rather than daily smokers. Women and Spanish speaking Hispanics were the most likely to be intermittent rather than former smokers. Attention to health media sources increased the likelihood that a person would be an intermittent smoker instead of a former or daily smoker. Believing that damage from smoking is avoidable and irreversible was associated with lower odds of being an intermittent smoker rather than a former smoker but did not differentiate intermittent smoking from daily smoking. CONCLUSION: The results indicate that tailoring smoking-cessation campaigns toward intermittent smokers from specific demographic groups by using health media may improve the effect of these campaigns and reduce social health disparities. PMID- 19288984 TI - Improved recognition of heart attack and stroke symptoms after a community-based intervention for older adults, Georgia, 2006-2007. AB - INTRODUCTION: In Georgia, mortality from stroke is 16% higher and from cardiovascular disease is 9% higher than it is nationally. Although 75% of Georgia adults have 2 or more modifiable risk factors for cardiovascular disease, less than half recognize all major heart attack and stroke warning symptoms. To reduce disability and prevent death from cardiovascular events, high-risk population groups should be able to recognize symptoms and seek immediate medical attention. METHODS: We evaluated a 4-month education intervention in 40 senior centers in Georgia. The intervention focused on improving knowledge of heart attack and stroke symptoms and on promoting lifestyle behaviors that prevent and manage cardiovascular disease and diabetes. Participants in a convenience sample completed a pretest questionnaire, the intervention, and a posttest questionnaire (N = 693, mean age, 75 years, 84% female, 45% black). RESULTS: After the intervention, recognition of all 5 symptoms of heart attack increased from 29% at the pretest to 46% at the posttest, and recognition of all 5 symptoms of stroke increased from 42% at the pretest to 65% at the posttest (for both conditions, P < .001). In linear regression analyses, independent positive predictors of change in knowledge were younger age and higher education. Most risk factors for cardiovascular disease were not predictive. CONCLUSION: The results of this evaluation provide an evidence base for the effectiveness of this intervention in improving knowledge about heart attack and stroke symptoms, which may translate to greater preparedness in these older adults for response to cardiovascular events. PMID- 19288985 TI - Area-level variation in adolescent smoking. AB - INTRODUCTION: The purpose of this study is to 1) examine the variability in the prevalence of adolescent smoking in 60 geographic areas of Minnesota and 2) assess how variability in area-level smoking prevalence is associated with area level sociodemographic characteristics. METHODS: Smoking data were collected from 3,636 adolescents residing in 60 areas of the state of Minnesota. Area-level characteristics were obtained from the 2000 US Census. Coefficient of variation was calculated to assess variability in smoking prevalence across areas, and mean smoking prevalence was compared above and below the median for each area-level characteristic. RESULTS: Substantial variation was found in adolescent smoking prevalence rates. Across the 60 areas, the percentage of adolescents that ever smoked varied from 13% to 53%, and the percentage of adolescents that smoked in the past 30 days ranged from 3% to 19%. Mean lifetime smoking prevalence was higher in areas with a higher percentage of residents with less than a high school education, a lower percentage of residents living in an urban area, lower median housing value and a lower median household income, a higher percentage of residents aged 16 years or older who were unemployed, and a higher percentage of residents with an income-to-poverty ratio less than 1.5. Similar results were found for past 30-day smoking prevalence among girls; however, no area-level characteristics were significantly associated with past 30-day smoking prevalence among boys. CONCLUSION: Results suggest that area-level characteristics may play an important role in adolescent smoking, particularly for girls. PMID- 19288986 TI - Parental perspectives on antismoking discussions with adolescents in rural African American households, May 2004-January 2005. AB - INTRODUCTION: The purpose of this study was to use qualitative interviews to examine antismoking discussions African American parents and adult family members have with adolescent children. This study is one of the first studies to examine the content of family discussions about not smoking among rural African American families from the perspective of parents and extended family members. METHODS: Interview topics included discussions with their children, how their children reacted to those discussions, expected and actual consequences for their children trying a cigarette, and perspectives on how best to keep their children from becoming cigarette smokers. A total of 72 African American households participated in the overall study, and 112 people were interviewed. RESULTS: Major themes that emerged included discussing the negative health and economic aspects of smoking and the influence of peer pressure. Likely consequences for trying a cigarette included talking to the child about the dangers of smoking and taking away privileges. Making cigarettes less accessible, continued discussions, leading by example, and not smoking around children were suggested as strategies to keep children from smoking. CONCLUSION: This study provides insight into antismoking socialization efforts in rural African American families and confirms that African American families are actively engaged in keeping their children from smoking. PMID- 19288987 TI - Use of complementary and alternative medicine among patients with arthritis. AB - INTRODUCTION: Previous studies suggest that people with arthritis have high rates of using complementary and alternative medicine (CAM) approaches for managing their arthritis, in addition to conventional treatments such as prescription medications. However, little is known about the use of CAM by diagnosis, or which forms of CAM are most frequently used by people with arthritis. This study was designed to provide detailed information about use of CAM for symptoms associated with arthritis in patients followed in primary care and specialty clinics in North Carolina. METHODS: Using a cross-sectional design, we drew our sample from primary care (n = 1,077) and specialist (n = 1,063) physician offices. Summary statistics were used to calculate differences within and between diagnostic groups, practice settings, and other characteristics. Logistic regression models clustered at the site level were used to determine the effect of patient characteristics on ever and current use of 9 CAM categories and an overall category of "any use." RESULTS: Most of the participants followed by specialists (90.5%) and a slightly smaller percentage of those in the primary care sample (82.8%) had tried at least 1 complementary therapy for arthritis symptoms. Participants with fibromyalgia used complementary therapies more often than those with rheumatoid arthritis, osteoarthritis, or chronic joint symptoms. More than 50% of patients in both samples used over-the-counter topical pain relievers, more than 25% used meditation or drew on religious or spiritual beliefs, and more than 19% used a chiropractor. Women and participants with higher levels of education were more likely to report current use of alternative therapies. CONCLUSION: Most arthritis patients in both primary care and specialty settings have used CAM for their arthritis symptoms. Health care providers (especially musculoskeletal specialists) should discuss these therapies with all arthritis patients. PMID- 19288988 TI - Risk and protective factors for tobacco use among 8th- and 10th-grade African American students in Virginia. AB - INTRODUCTION: Few studies have simultaneously examined the influence of multiple domains of risk and protective factors for smoking among African Americans. This study identified individual-peer, family, school, and community risk and protective factors that predict early cigarette use among African American adolescents. METHODS: Data from 1,056 African American 8th and 10th graders who completed the 2005 Community Youth Survey in Virginia were analyzed by using logistic regression. RESULTS: The prevalence of smoking among the weighted sample population was 11.2%. In univariate analyses, the strongest predictors of smoking were low academic achievement, peer drug use, and early substance use (individual domain). In multivariate analyses, these factors and being in the 10th grade were significant predictors. The single protective factor in multivariate analyses was in the school domain (rewards for prosocial behavior in the school setting). When family and community variables were entered into a model in which individual-peer and school factors were controlled for, these variables were not significantly associated with smoking, and they failed to improve model fit. CONCLUSION: These findings suggest that tobacco prevention programs that aim to increase school connectedness while decreasing youth risk behaviors might be useful in preventing cigarette use among African American adolescents. Given the relative importance of peer drug use in predicting smoking among African American youth, more work is needed that explores the accuracy of youths' perceptions of their friends' cigarette use and how family factors may moderate this risk. PMID- 19288989 TI - Caregivers of older adults with cognitive impairment. AB - INTRODUCTION: Because of the growing number of caregivers and the awareness of related health and quality-of-life issues, caregiving has emerged as an important public health issue. We examined the characteristics and caregiving experiences of caregivers of people with and without cognitive impairment. METHODS: Participants (n = 668) were adults who responded to the 2005 North Carolina Behavioral Risk Factor Surveillance System. Caregivers were people who provided regular care to a family member or friend aged 60 years or older either with or without cognitive impairment (ie, memory loss, confusion, or Alzheimer's disease). RESULTS: Demographic characteristics of caregivers of people with cognitive impairment were similar to those of caregivers of people without cognitive impairment. However, compared with caregivers of people without cognitive impairment, caregivers of people with cognitive impairment reported higher levels of disability, were more likely to be paid, and provided care for a longer duration. Care recipients with cognitive impairment were more likely than care recipients without cognitive impairment to be older, have dementia or confusion, and need assistance with memory and learning. CONCLUSION: State-level caregiving surveillance is vital in assessing and responding to the needs of the growing number of caregivers. PMID- 19288990 TI - Low awareness of overweight status among parents of preschool-aged children, Minnesota, 2004-2005. AB - INTRODUCTION: Many studies have found that parents of overweight children do not perceive their child to be overweight. Little is known, however, about the extent to which such misperceptions exist among parents of preschool-aged children. METHODS: We analyzed data that were collected in 2004-2005 from parents of 593 preschool-aged children in 20 child care centers in the Minneapolis-St. Paul, Minnesota, metropolitan area. Parents were asked how they would classify their preschooler's weight, and children's height and weight were measured. RESULTS: Of the predominantly white, educated sample, most parents (90.7%) of overweight preschoolers classified their child as normal weight. An even higher percentage (94.7%) of children at risk for overweight were classified as normal weight by their parents. Most parents of normal-weight children classified their child's weight as average. However, 16.0% classified their normal-weight child as underweight or very underweight. CONCLUSION: Results indicate that parents are unlikely to recognize childhood overweight among preschool-aged children, which is concerning because parents of overweight children may be unlikely to engage in obesity prevention efforts for their child if they do not recognize their child's risk status. A notable proportion of parents of normal-weight children perceived their child to be underweight, which suggests that parents of normal-weight children may be more concerned with undernutrition than overnutrition. PMID- 19288991 TI - Prevalence of self-reported prediabetes among adults participating in a community based health awareness program, New York State. AB - INTRODUCTION: The purpose of this study was to assess crude, age-adjusted, and risk-factor-specific prevalences of self-reported prediabetes and to identify factors associated with self-reported prediabetes in an adult population. METHODS: Data were collected through questionnaires completed by a racially diverse sample of diabetes-free adult participants in the statewide community based wellness and diabetes awareness program in New York State during 2006 (N = 2,572). Prediabetes was determined by the affirmative answer to the question, "Have you ever been told by a doctor that you have prediabetes?" RESULTS: The overall crude prevalence of self-reported prediabetes was 9.1%, and the age adjusted prevalence was 7.6%. The age-adjusted prevalence of prediabetes was significantly lower among non-Hispanic blacks (4.2%) and significantly higher among American Indians (22.4%), compared with the prevalence among non-Hispanic whites (7.3%). The prevalence of self-reported prediabetes was uniformly higher among older (aged >or=45 years) adults than younger (aged <45 years) adults, overall and in each racial/ethnic group. In all age and racial/ethnic groups, the prevalence significantly increased with the number of additional risk factors. The best fit multivariate logistic regression model identified that self-reported prediabetes was associated with family history of diabetes (odds ratio [OR], 3.65), body mass index 25.0 kg/m(2) or higher (OR, 2.79), age 45 years or older (OR, 2.77), and having health insurance (OR, 2.38). CONCLUSION: This study found that adults who were at high risk for diabetes and had health insurance were more likely to report having prediabetes. Community-based diabetes prevention needs to consider strategies to increase detection of prediabetes in high-risk uninsured people and to raise general awareness of prediabetes. PMID- 19288992 TI - HPV vaccine attitudes and practices among primary care providers in Appalachian Pennsylvania. AB - INTRODUCTION: The incidence of cervical cancer in Appalachia exceeds the national rate; rural Appalachian women are at especially high risk. We assessed the attitudes and practices related to human papillomavirus vaccination among providers in primary care practices in a contiguous 5-county area of Appalachian Pennsylvania. METHODS: In December 2006 and May 2007, all family medicine, pediatric, and gynecology practices (n = 65) in the study area were surveyed by 2 faxed survey instruments. RESULTS: Of the 65 practices, 55 completed the first survey instrument. Of these 55, 44 offered the vaccine to their patients. Forty of the 44 practices offered it to girls and women aged 9 to 26 years, and 11 were willing to accept referrals from other practices for vaccination. The average reported charge for each of the 3 required injections was $150. Of the 55 practices that responded to the first survey instrument, 49 responded to the second survey instrument, 46 of which recommended the vaccine to their patients. CONCLUSION: The prevalence of offering the vaccine against human papillomavirus was high in this area of Appalachian Pennsylvania. Future interventions may focus on community education because the vaccine is available from most providers. PMID- 19288993 TI - Acculturation and dental visits among Hispanic adults. AB - INTRODUCTION: Acculturation may strongly influence use of or access to health services among Hispanics in the United States. We assessed the relationships between acculturation and use of oral health services among Hispanic adults in the United States. METHODS: Data were analyzed from Hispanic adults aged 18 years or older who participated in the 2006 Behavioral Risk Factor Surveillance System. Hispanics were defined by self-report of Spanish or Hispanic heritage. Preference to be interviewed in English or Spanish was used as a proxy for acculturation. Having had a dental visit in the previous 12 months was used as a proxy for use of oral health services. RESULTS: English-speaking Hispanics were more likely to have had a dental visit in the previous 12 months compared with Spanish-speaking Hispanics (crude odds ratio [OR], 1.52; 95% confidence interval [CI], 1.36-1.71). After controlling for potential confounders, language was not significantly associated with having had a dental visit (OR, 1.05; 95% CI, 0.87-1.26; P = .61,). The most significant predictors for having had a dental visit in the previous 12 months were sex, education, income, and having health insurance. CONCLUSION: Acculturation assessed by language spoken was not significantly associated with having had a dental visit in the previous 12 months among adult Hispanics in the United States. The common determinants of health care use, such as sex, income, level of education, and health insurance status, were the most significant predictors of use of oral health services among adult Hispanics. PMID- 19288994 TI - Body weight dissatisfaction among Israeli Jewish and Arab women with normal or overweight-obese body mass index, Israeli INHIS-1, 2003-2004. AB - INTRODUCTION: In Israel, 58.9% of Jewish and Arab Israeli women aged 25 to 64 years are overweight or obese (body mass index >or=25 kg/m(2)). The objective of this analysis is to describe body weight dissatisfaction differences between Jewish and Arab Israeli women with normal or overweight-obese body mass index. METHODS: This analysis included 1,393 Jewish and Arab women who participated in the Israeli National Health Interview Survey, 2003-2004. The survey covered a random sample of the Israeli general population aged 21 years or older. All variables were based on self-report. Body weight dissatisfaction was a multiple choice question in the survey that offered the following responses: very satisfied, satisfied, reasonably satisfied, not satisfied, or very unsatisfied. Univariate and multivariate analyses were conducted. RESULTS: Overall, 39.1% of Jewish women reported body weight dissatisfaction, compared with 29.1% of Arab women. Older overweight-obese Arab women had a lower prevalence of body weight dissatisfaction than Jewish women of the same age group, which indicates cultural differences in body weight dissatisfaction among older overweight-obese women. However, cultural differences do not appear to influence body weight dissatisfaction among younger Jewish and Arab women of normal weight. CONCLUSION: This study suggests that Jewish and Arab women differ in their perceptions of body weight. Interventions tailored to each group are needed to promote healthy dietary and physical activity behaviors. PMID- 19288995 TI - Secondary prevention of myocardial infarction with nonpharmacologic strategies in a Medicaid cohort. AB - BACKGROUND: The quality of health care after myocardial infarction (MI) may be lacking; in particular, guidelines for nonpharmacologic interventions (cardiac rehabilitation, smoking cessation) may receive insufficient priority. We identified gaps between secondary prevention guidelines and ambulatory care received by Medicaid enrollees after an MI. METHODS: MI survivors were selected by using 2004 Washington State Medicaid administrative claims. Deidentified data were abstracted for hospitalizations, ambulatory care, and prescriptions for 365 days after the MI. Cox regression analysis compared utilization of guideline directed secondary prevention strategies with death and recurrent hospitalization. RESULTS: The sample size was 372. Fifty patients died during the year after the MI, and 144 were rehospitalized. Only 2 patients attended a cardiac rehabilitation program. Tobacco cessation counseling was associated with a 66% reduction in death, but only 72.6% of smokers were counseled. Less than half (45.4%) of patients saw a primary care provider within 90 days of their MI, and 7.5% never contacted a health care provider. Receiving regular primary care was associated with a decreased risk for death (hazard ratio, 0.91; 95% confidence interval, 0.84-0.97, P < .01). A protective trend was associated with care by a cardiologist, but only 21.5% received specialist care. CONCLUSION: Analysis of Medicaid claims data suggests rates of secondary prevention are less than optimal. To improve survival and reduce rehospitalization after an MI, policy changes (tobacco cessation benefits, expansion of rehabilitation programs), health care capacity (training, referral patterns, and coordination of care), and improvements to access (removing barriers, increasing facilities, targeting minority populations) could be implemented. PMID- 19288996 TI - Differences in patterns of alcohol consumption among Hispanics in the United States, by survey language preference, Behavioral Risk Factor Surveillance System, 2005. AB - INTRODUCTION: Alcohol consumption is pervasive in the United States, and extent of alcohol consumption for the growing US Hispanic population needs further study. We examined the association between language chosen for a national health survey and alcohol use among Hispanic adults. METHODS: Hispanic participants aged 18 years and older (N = 20,234) from the 2005 Behavioral Risk Factor Surveillance System were stratified by choice of language (English, n = 13,035; Spanish, n = 7,199) for completing the survey. Differences for these 2 groups in current alcohol use, heavy alcohol use, and binge drinking were determined by using chi2 analyses and logistic regression models. RESULTS: In bivariate associations, current drinking (P < .001), heavy drinking (P < .001), and binge drinking (P = .002) were significantly higher among participants who chose to complete the survey in English than among those who elected to complete the survey in Spanish. After controlling for demographic characteristics, associations between language choice and drinking behaviors were found to be greatest among women. Compared with women who chose to complete the survey in Spanish, women who chose to complete the survey in English were more than twice as likely to report current drinking (odds ratio [OR] = 2.42, 95% confidence interval [CI] = 2.02-2.91), heavy drinking (OR = 3.82, 95% CI = 1.44-10.10), and binge drinking (OR = 2.51, 95% CI = 1.64-3.84). CONCLUSION: This study suggests that language choice when completing a health survey is a predictor of high levels of alcohol use among Hispanic adults in the United States and that differences in drinking behaviors based on language choice for a survey are more profound among women. PMID- 19288997 TI - Family history of diabetes: exploring perceptions of people at risk in the Netherlands. AB - INTRODUCTION: The aim of this study was to explore the perceptions of causes, risk, and control with regard to diabetes and the role of family history among people at increased risk for type 2 diabetes. METHODS: Semistructured interviews were conducted among people aged 57 to 72 years with (n = 9) and without (n = 12) a family history of diabetes. RESULTS: Participants mentioned different causes for diabetes; these were often a combination of genetic and behavioral factors. Some participants with a family history expressed incoherent causal beliefs; their general ideas about the causes of diabetes did not explain why their relatives were affected. The role of genetics as a cause for diabetes was more pronounced when people perceived diabetes as "running in the family," and this finding did not necessarily relate to a high number of affected relatives. Although people with a family history were aware of the diabetes in their family, they did not always associate their family history with increased risk, nor did they worry about getting diabetes. The absence of diabetes in the family was often used as a reason to perceive a low risk. Participants who primarily perceived genetic predisposition as a cause felt less able to prevent getting diabetes. CONCLUSION: Future diabetes prevention strategies would benefit from giving more attention to individual perceptions, especially in the context of family history, explaining the multifactorial character of diabetes, and highlighting effective ways to reduce the risk. PMID- 19288998 TI - Cholesterol screening by marital status and sex in the United States. AB - INTRODUCTION: Marital status may be a predisposing factor related to preventive health screenings, which may in part explain the "healthy marriage" effect. This study investigates differences in the likelihood of being screened for cholesterol by marital status for men and women. METHODS: Medical Expenditure Panel Surveys from 2003 through 2005 were used to calculate the likelihood of self-reported cholesterol screening in the past year by marital status and sex. Several rounds of interviews during a 2-year period resulted in a sample of 36,594 US adults. RESULTS: Most married, widowed, and divorced/separated people reported cholesterol screening in the past year. The highest percentages of people being screened for cholesterol were widowed men (75%) and women (81%). By contrast, 26% of single men and 38% of single women reported cholesterol screening. In multivariate models, being unmarried was associated with lower odds of cholesterol screening among men and women. The lowest likelihood of screening was associated with widowed status for both men (odds ratio, 0.56) and women (odds ratio, 0.53). CONCLUSION: Marital status is a predisposing factor for cholesterol screening. Public health interventions aimed at improving preventive screening should focus on social networks, especially family members. PMID- 19288999 TI - The association between body mass index and arthritis among US adults: CDC's surveillance case definition. AB - INTRODUCTION: The Centers for Disease Control and Prevention modified the surveillance case definition of arthritis to a more stringent form in 2002. To date, the association between arthritis and obesity (an established risk factor for arthritis) has not been examined with the new definition. We describe the association between body mass index (BMI) (kg/m(2)) and arthritis using the new arthritis case definition to provide a more accurate assessment of the relationship between weight and arthritis among US adults. METHODS: We used data from the 2005 Behavioral Risk Factor Surveillance System (N = 356,112) and univariate and multivariate analyses to assess the relationship between BMI and arthritis among US adults. RESULTS: Overall, 26% of US adults had self-reported arthritis. Obese respondents (BMI >or=30.0 kg/m(2)) were 1.9 times more likely to report arthritis compared with normal-weight respondents (BMI <25.0 kg/m(2)), and distinguishing between obese levels revealed an even greater association between BMI and arthritis (class III obesity [BMI >or=40.0], odds ratio [OR] = 3.3, 95% confidence interval [CI] = 3.1-3.6; class II obesity [BMI 35.0-39.9 kg/m2], OR = 2.5, 95% CI = 2.3-2.7; class I obesity [BMI 30.0-34.9], OR = 1.9, 95% CI = 1.8 2.0). CONCLUSION: BMI is an independent risk factor for self-reported arthritis. Maintaining a healthy weight may delay the onset of arthritis. More research is needed to determine the effect of weight loss on the progression of arthritis in overweight individuals. PMID- 19289000 TI - Novel approach, using end-of-life issues, for identifying items for public health surveillance. AB - Using end-of-life (EOL) issues to provide context, we introduce a novel approach to identify potential items for public health surveillance. Our method involved an environmental scan of existing EOL surveys and included the following steps: 1) consulting experts for advice on critical EOL topics, 2) identifying a broad sample of EOL surveys, and 3) using an abstraction tool to characterize surveys and survey items. We identified 36 EOL surveys; of these, 10 were state-based surveys. Of the 1,495 EOL items (range, 4 to 126 items per survey), 333 items could be classified in 1 of 11 topic areas of interest. Information on the surveys and these 333 items was entered into a database. As a result of this process, we identified topics for which many EOL items already exist and topics for which items should be developed. We describe the value of this approach and potential next steps for our project. PMID- 19289001 TI - Wisconsin's health department-university partnership model for comprehensive cancer control. AB - Cancer causes substantial morbidity and mortality every year in the United States. To address cancer prevention and control in Wisconsin, the Wisconsin Division of Public Health and the University of Wisconsin Paul P. Carbone Comprehensive Cancer Center forged a unique partnership. Using funds from the state legislature, the university, and the Centers for Disease Control and Prevention, the Wisconsin Comprehensive Cancer Control Program was created. This health department-university partnership model has allowed both institutions to contribute their distinct strengths to projects that neither organization would have been able to complete on its own. Some challenges also have arisen during development and execution of the program, but overall, this collaborative partnership has brought diverse groups together to develop and implement evidence based cancer control programs and policies in Wisconsin. PMID- 19289002 TI - Family physicians as team leaders: "time" to share the care. AB - A major contributor to shortfalls in delivery of recommended health care services is lack of physician time. On the basis of recommendations from national clinical care guidelines for preventive services and chronic disease management, and including the time needed for acute concerns, sufficiently addressing the needs of a standard patient panel of 2,500 would require 21.7 hours per day. The problem of insufficient time indicates that primary care requires broad, fundamental changes. The creation of primary care teams that include members such as physician assistants, nurse practitioners, dietitians, health educators, and lay coaches is important to meeting patients' primary care needs. PMID- 19289003 TI - A comprehensive worksite wellness program in Austin, Texas: partnership between Steps to a Healthier Austin and Capital Metropolitan Transportation Authority. AB - BACKGROUND: In 2003, Steps to a Healthier Austin was funded by the Centers for Disease Control and Prevention to implement chronic disease prevention and health promotion activities. We report Steps to a Healthier Austin's partnership with Health & Lifestyles Corporate Wellness, Inc (Health & Lifestyles), to provide a worksite wellness program for Capital Metropolitan Transportation Authority (Capital Metro), Austin's local transit authority. CONTEXT: Capital Metro employs 1,282 people. In 2003, Health & Lifestyles was hired to help promote healthier lifestyles, increase employee morale, and combat rising health care costs and absenteeism rates. METHODS: Health & Lifestyles provided consultations with wellness coaches and personal trainers, a 24-hour company fitness center, personalized health assessments, and preventive screenings. The program expanded to include healthier food options, cash incentives, health newsletters, workshops, dietary counseling, smoking cessation programs, and a second fitness center. CONSEQUENCES: Participants in the wellness program reported improvements in physical activity, healthy food consumption, weight loss, and blood pressure. Capital Metro's total health care costs increased by progressively smaller rates from 2003 to 2006 and then decreased from 2006 to 2007. Absenteeism has decreased by approximately 25% since the implementation of the program, and the overall return on the investment was calculated to be 2.43. INTERPRETATION: Since the implementation of the wellness program in 2003, Capital Metro has seen a reduction in costs associated with employee health care and absenteeism. PMID- 19289004 TI - Engaging employers to develop healthy workplaces: the WorkWell initiative of Steps to a Healthier Washington in Thurston County. AB - BACKGROUND: The WorkWell initiative of Thurston County, Washington, established by Steps to a Healthier Washington in Thurston County (Thurston County Steps), focuses on recognizing and supporting local employers who make a commitment to address workforce health issues by implementing programs within their organizations to help adults reach Healthy People 2010 objectives. This article reports on the WorkWell initiative and resulting WorkWell program. CONTEXT: The WorkWell initiative was developed to address the needs of private and public employers in Thurston County, Washington, to reduce the prevalence of chronic diseases through policy, practice, and environmental changes. METHODS: Thurston County Steps recruited local employers to participate in advisory work groups to identify healthy workplace interventions that would be feasible for the employers and initiate a shift in organizational culture. The WorkWell initiative developed 2 distinct approaches - 1 for private sector (designation program) and another for public sector (action planning). CONSEQUENCES: Twenty-six employers with approximately 4,700 employees were recognized with WorkWell Healthy Workplace designations for implementing changes that included encouraging stairwell use, providing low- or no-cost healthy meals for employees, and providing healthy foods at meetings. Four public agencies with approximately 4,400 employees have participated in an assessment and action planning process to help government employers focus their efforts and resources to support workforce health promotion. INTERPRETATION: Unique partnerships between Thurston County Steps and other employers, private and public, demonstrate the important role employers can play in reducing chronic disease to improve a community's overall health. PMID- 19289005 TI - Building company health promotion capacity: a unique collaboration between Cargill and the Centers for Disease Control and Prevention. AB - BACKGROUND: The US Centers for Disease Control and Prevention (CDC) helps protect the health and safety of all people. The workplace can be used to reach millions of workers and their families with programs, policies, and benefits that promote health. We describe a CDC-led project to build Cargill's workplace health promotion capacity and identify the importance of a company liaison in the public private relationship. CONTEXT: The project goals were to engage diverse Cargill personnel, conduct a workplace health assessment, aid in the development of a workplace health program action plan, and develop Cargill's internal capacity using knowledge and skill-building. METHODS: CDC partnered with Cargill on a workplace health promotion project to build Cargill's capacity. A multicomponent assessment was conducted to determine priority employee health issues, stakeholder meetings were held to engage and educate Cargill management and employees, and technical assistance was provided regularly between CDC and Cargill. CONSEQUENCES: Identifying a company liaison to work with an external assessment team is critical to building capacity for a successful workplace health project. This relationship creates an understanding of company culture and operations, facilitates access to key stakeholders and data, and provides opportunities to enhance capacity and sustainability. INTERPRETATION: Employers undertaking workplace health promotion projects should identify a senior-level person to serve as the company health leader or liaison and who can devote the time necessary to build trusting relationships with partners to ensure project success. This person is valuable in facilitating communications, data collection, logistical support, troubleshooting, and influencing employer workplace health practices. PMID- 19289006 TI - Healthy Maine Partnerships: policy and environmental changes. AB - BACKGROUND: Tobacco settlement funds were used to establish the Healthy Maine Partnerships (HMPs) to reduce tobacco use, increase physical activity, and improve nutrition through local policy and environmental change. CONTEXT: The HMP model is a progressive approach to public health. It provides for coordinated efforts between state and local partners for health promotion and disease prevention. Community coalitions, supported with funding and guidance by the state, are the basis for policy and environmental change. METHODS: The state awarded contracts and provided program guidance to foster policy and environmental change at the local level. The partnerships' efforts were assessed with a retrospective evaluation that consisted of 2 data collection periods conducted using the same tool. A survey booklet containing lists of possible environmental and policy changes was developed and mailed - once in 2005 and once in 2006 - to all 31 local partnership directors and school health coordinators who completed it. Additional data were collected from the local partnerships in the form of narrative reports required by their funder (Maine Center for Disease Control and Prevention). CONSEQUENCES: All local partnerships implemented policy or environmental interventions to address tobacco use, physical activity, and nutrition during the period covered by the surveys (July 2002-June 2005 [fiscal years 2003-2005]). Cumulatively, more than 4,600 policy or environmental changes were reported; tobacco use policies represent most changes implemented. A second round of HMP funding has since been secured. INTERPRETATION: Although the survey methodology had limitations, results suggest that much work has been accomplished by the local partnerships. Plans are to share success stories among partnerships, provide training, and continue to improve the public health infrastructure in Maine. PMID- 19289007 TI - Evaluating partnerships to prevent and manage chronic disease. AB - To be effective and sustain themselves over time, public-private partnerships must make evaluation a priority. Specifically, partnerships should evaluate 1) their infrastructure, function, and processes; 2) programs designed to achieve their mission, goals, and objectives; and 3) changes in health and social status, organizations, systems, and the broader community. This article describes how to 1) develop a comprehensive evaluation strategy based on partnership theory; 2) select short-term, intermediate, and long-term indicators to measure outcomes; 3) choose appropriate methods and tools; and 4) use evaluation results to provide accountability to stakeholders and improve partnership function and program implementation. PMID- 19289008 TI - Development of a community-wide cardiovascular risk reduction assessment tool for small rural employers in upstate New York. AB - Employers are implementing workplace health promotion programs that address modifiable health risk factors such as overweight and obesity, smoking, high blood pressure, high cholesterol, physical inactivity, poor diet, and high stress. Research with large employers has found that these programs can improve workers' health and decrease the costs associated with medical care, absenteeism, and presenteeism. Despite their promise, health promotion programs are not widely embraced by small businesses, especially those in rural communities. This article reviews the barriers encountered by small and rural businesses in implementing health promotion programs. We describe an approach developed in cooperation with the New York State Department of Health's Healthy Heart Program and the Cayuga Community Health Network to engage small businesses in health promotion. We review the development and implementation of an assessment tool created to evaluate current workplace health promotion programs, policies, and practices targeting cardiovascular disease among small, rural employers in upstate New York. Potential benefits of the assessment tool are discussed, and the instrument is made available for the public. PMID- 19289009 TI - Development and status of the National Oral Health Surveillance System. AB - During the last 2 decades of the 20th century, few national, state, or local oral health programs were able to conduct public health surveillance in a timely fashion. Under the leadership of the Association of State and Territorial Dental Directors and with substantial support from the Division of Oral Health at the Centers for Disease Control and Prevention, the National Oral Health Surveillance System was established as a first step in helping oral health programs routinely document population needs and program impact with standard, feasible methods. In 1999, the Council of State and Territorial Epidemiologists approved 7 oral health indicators for public health surveillance: 3 for adults (most recent dental visit, most recent dental cleaning, total tooth loss) using data from the Behavioral Risk Factor Surveillance System; 3 for third-grade students (presence of treated or untreated dental caries, untreated tooth decay, dental sealants) collected by states using a standard screening protocol; and the percentage of the population served by public water systems that receives optimally fluoridated water, tracked through the Water Fluoridation Reporting System. The Web site that describes the National Oral Health Surveillance System (http://www.cdc.gov/nohss/) and provides access to current indicators was launched in 2001 with adult and water fluoridation data for all states; child indicators were added later. Data are now available electronically for 35 to 51 states (including the District of Columbia), depending on the indicator, indicating progress toward state-specific monitoring of these oral health indicators. PMID- 19289010 TI - An emergency medical services toolkit for improving systems of care for stroke in North Carolina. AB - The Centers for Disease Control and Prevention is partnering with the National Association of Chronic Disease Directors and the North Carolina Office of EMS to design, develop, and implement an emergency medical services (EMS) performance improvement toolkit to evaluate opportunities to improve the emergency identification and treatment of acute stroke. The EMS Acute Stroke Care Toolkit is being developed, tested, and implemented in all 100 counties in the state by the EMS Performance Improvement Center, the agency that provides technical assistance for EMS in North Carolina. The toolkit helps each EMS system in defining, measuring, and analyzing their system of care and promotes collaboration through public education, regional stroke planning with hospitals, EMS service configuration, EMS staffing patterns, EMS education, and timely care delivery. We outline the issues surrounding acute stroke care, the role of emergency medical systems in stroke care, and the components of the EMS Acute Stroke Care Toolkit designed to improve EMS systems and outcomes for stroke patients. PMID- 19289011 TI - Culturally tailored cancer communication, education, and research: the highways and back roads of Appalachia. AB - We seek to start a dialogue about the challenges cancer control researchers and specialists may face in attempting to understand the Appalachians' experience with cancer. Through examples drawn from our own research among Appalachian communities, we discuss the hazards of defining a culture in order to develop culturally tailored cancer control interventions and programs. We also acknowledge that cancer control work in Appalachia requires "cultural mapping," highlighting cultural beliefs, norms, and realities that may be linked to cancer mortality and morbidity. Although cancer control specialists and researchers have to rely on cultural maps, they must also remain critical of such maps. Subsequently, we describe a mapping approach around the metaphor of "signposts," directional indicators that point to broad cultural attributes but do not reduce the culture to a narrow set of traits. The interplay of these signposts ultimately helps cancer educators, communicators, and researchers better understand authentic Appalachia. PMID- 19289012 TI - The private partners of public health: public-private alliances for public good. AB - OBJECTIVE: We sought to convey lessons learned by the Centers for Disease Control and Prevention's (CDC's) Prevention Research Centers (PRCs) about the value and challenges of private-sector alliances resulting in innovative health promotion strategies. Several PRCs based in a variety of workplace and community settings contributed. METHODS: We conducted interviews with principal investigators, a literature review, and a review of case studies of private-sector alliances in a microbusiness model, a macrobusiness model, and as multiparty partnerships supporting public health research, implementation, and human resource services. RESULTS: Private-sector alliances provide many advantages, particularly access to specialized skills generally beyond the expertise of public health entities. These skills include manufacturing, distribution, marketing, business planning, and development. Alliances also allow ready access to employee populations. Public health entities can offer private-sector partners funding opportunities through special grants, data gathering and analysis skills, and enhanced project credibility and trust. Challenges to successful partnerships include time and resource availability and negotiating the cultural divide between public health and the private sector. Critical to success are knowledge of organizational culture, values, mission, currency, and methods of operation; an understanding of and ability to articulate the benefits of the alliance for each partner; and the ability and time to respond to unexpected changes and opportunities. CONCLUSION: Private-public health alliances are challenging, and developing them takes time and resources, but aspects of these alliances can capitalize on partners' strengths, counteract weaknesses, and build collaborations that produce better outcomes than otherwise possible. Private partners may be necessary for program initiation or success. CDC guidelines and support materials may help nurture these alliances. PMID- 19289013 TI - Severe weather, band practice, coal trucks, and other real-world experiences in conducting focus group research in central Appalachia. PMID- 19289014 TI - American Heart Association's behavioral roundtable for preventable disparities. PMID- 19289015 TI - A case for public-private partnerships in health: lessons from an honest broker. PMID- 19289016 TI - The academy's pivotal role in supporting public-private partnerships to prevent chronic diseases. PMID- 19289017 TI - Health care reform hinges on private-sector collaboration. AB - America's health care system is characterized by rising costs, increasing numbers of Americans who lack health insurance coverage, and poor quality of health care delivery. The convergence of these factors is adversely affecting not only the health of Americans but also the ability of businesses to compete successfully in a global marketplace. AARP and other nonprofit organizations are collaborating with the private sector to have more people covered by health insurance and to educate them to make behavioral choices that prevent chronic disease and ultimately lower costs. PMID- 19289018 TI - Medicine, big business, and public health: wake up and smell the Starbucks. AB - The provision of ambulatory care by major retailers is small but growing, providing speedy attention to consumers with minimal wait times and no appointments necessary. Users of these clinics are satisfied with the care they receive. Primary care physicians have opposed retail clinics, concerned that conditions will be misdiagnosed, opportunities to address comorbidities and risk behaviors will be missed, necessary follow-up care will be delayed or absent, and the profit motive will lead to cutting corners. Public health is now being challenged to capitalize on the advantageous possibilities these clinics can offer, such as serving uninsured patients, while remaining vigilant regarding potential hazards, such as financial pressures that could negatively affect health care quality, continuity, and accessibility. PMID- 19289019 TI - Are partnerships with the tobacco industry and food and beverage industry possible? An interview with Michael Eriksen by Elizabeth Majestic. PMID- 19289020 TI - New models for partnering with the business sector: an interview with Gene Matthews by Elizabeth Majestic. PMID- 19289021 TI - Projected burden of chronic, noncommunicable diseases in Jordan. PMID- 19289022 TI - Treatment of varicose veins. AB - Varicose veins (VVs) are the most common manifestation of chronic venous insufficiency, affecting 25% of women and 15% of men. Reticular veins and telangiectasias (spider veins) are found in more than 80% of the general population. VVs produce symptoms of pain, swelling, heaviness, fatigue, and pruritus and predispose patients to complications including bleeding, superficial thrombophlebitis, and ulcerations that interfere with activities of daily living and result in lost time from work. Current treatments for VVs include conservative measures, and when these are unsuccessful, more invasive surgical and endovenous interventions primarily aimed at reducing venous hypertension and preventing progression to chronic inflammation and ulcerations. Surgical procedures including saphenous vein stripping, ligation of the saphenofemoral junction, and ambulatory phlebectomy are effective in the treatment of VVs but are associated with a high complication rate and recovery time. Emerging endovenous therapies, including endovenous laser therapy, radiofrequency ablation, and endovenous foam sclerotherapy, have shown similar efficacy in the treatment of VVs compared with more invasive surgical procedures, with lower complication rates and less time lost from work. PMID- 19289023 TI - Venous thromboembolism in pregnancy. AB - Low molecular weight heparins (LMWHs) appear to be as safe and effective as unfractionated heparin (UFH) for venous thromboembolic disease (VTED) treatment or prophylaxis during pregnancy. Experience with other parenteral anticoagulant drugs is very limited, and no alternative oral anticoagulants are available to date. In addition to cost, challenges of long-term LMWH use during pregnancy that have not been addressed by controlled clinical trials include a) ideal dosing as pregnancy advances, b) the need for LMWH monitoring by anti-Xa activity levels, and c) ideal therapeutic management as the delivery date nears. Because therapeutic-intensity anticoagulation during pregnancy is challenging, many practitioners favor a more "aggressive" approach toward VTED prophylaxis in women perceived to be at very high risk of thrombosis during pregnancy. Best evidence to date suggests that most women with thrombophilias or with a previous "situational" VTED event probably do not require VTED prophylaxis antepartum, but postpartum anticoagulation prophylaxis is recommended for a few weeks. For those with a history of previous idiopathic VTED or VTED associated with "hormonal challenge" (such as with contraceptive use or previous pregnancy), prophylaxis beginning antepartum may be considered and discussed with the patient. Selected cases of "severe" thrombophilia are probably best managed by initiation of pharmacologic VTED prophylaxis antepartum. However, it must be emphasized that data from prospective controlled clinical trials are lacking. PMID- 19289024 TI - Thrombophilia: 2009 update. AB - As venous thrombosis is mostly caused by disturbances in the plasma coagulation system, abnormalities of coagulation factors are mostly risk factors for venous thromboembolism (VTE). Relatively little is known about thrombophilias that predispose to arterial thromboembolism. Although some abnormalities in the fibrinolytic pathway appear to predispose to arterial thrombosis, the associations are weak and often inconsistent between studies. At present, there is not enough consistent and clinically meaningful information to include fibrinolytic parameters in a clinical thrombophilia workup. Controversy exists as to which patients and family members to test for thrombophilia. Several testing guidelines exist. Routine screening for inherited thrombophilias is not indicated in patients with VTE provoked by immobility, surgery, and malignancy, or in those with arterial thrombosis with arteriosclerosis risk factors. Heterozygous factor V Leiden (FVL) and prothrombin 20210 mutations increase the risk for recurrent VTE only slightly once anticoagulation is stopped. Therefore, decisions regarding the length of anticoagulant therapy typically are not influenced by finding one of these heterozygous mutations. The main reason to perform thrombophilia testing in a patient is to detect a strong thrombophilia (ie, antithrombin deficiency, antiphospholipid antibody syndrome, homozygous FVL, double-heterozygous FVL plus prothrombin 20210 mutation, protein C deficiency, and maybe protein S deficiency). The finding of a strong thrombophilia has several clinical consequences: it decreases the threshold to recommend long-term anticoagulation in a patient with unprovoked VTE; facilitates discussion regarding whether anticoagulant or antiplatelet therapy is the preferred empiric treatment for a patient who had an unexplained arterial, nonarteriosclerotic thromboembolic event; and leads to the consideration of testing asymptomatic female family members for the identified thrombophilia(s) so they can be counseled on their risk of thromboembolism, the use of hormonal therapies, and the potential benefit of pre- and postpartum anticoagulant therapy. PMID- 19289025 TI - Thrombolytic therapy for deep vein thrombosis. AB - The incorporation of endovascular interventions into mainstream clinical practice for patients with deep vein thrombosis (DVT) has been changing rapidly as a result of four major trends. First, there now is a much greater appreciation among practicing physicians of the major impact of the postthrombotic syndrome (PTS) on DVT patients' late quality of life. Second, there have been rapid advances in the methods by which thrombus can be rapidly removed using endovascular techniques, resulting in greater efficiency and treatment safety. Third, these changes together have spurred a major change in recommended DVT treatment paradigms: there is now published expert consensus that adjunctive endovascular DVT thrombolysis should be strongly considered as a first-line treatment strategy for selected patients with extensive acute proximal DVT. Finally, DVT researchers from multiple subspecialties have finally come together in a major multidisciplinary clinical trial project to subject endovascular DVT thrombolysis to the rigorous scientific testing that is needed for clinicians to be confident in its efficacy, safety, and cost-effectiveness as a first-line DVT treatment strategy. Pending the results of this and other studies, medical physicians should routinely inform DVT patients about the long-term consequences of DVT; ensure that they understand the importance of anticoagulant therapy and elastic compression stockings in preventing PTS; provide a balanced discussion of the risks, benefits, and uncertainties associated with endovascular DVT thrombolysis to patients with extensive proximal DVT; and develop collaborative clinical and research relationships with physicians experienced with endovascular DVT therapy. PMID- 19289026 TI - Endovascular treatment of aortic aneurysms: state of the art. AB - Abdominal aortic aneurysms (AAAs) continue to be a leading cause of death, with increasing incidence and prevalence. Endovascular aneurysm repair (EVAR) now represents the most common method of AAA repair in the United States. Ongoing improvements in endovascular stent-graft technology have occurred since the first published report of EVAR in 1991. These improvements have led to multiple US Food and Drug Administration-approved devices, streamlined operative techniques, and extended applicability of EVAR. Despite these facts, basic anatomic considerations still eliminate many patients from being offered EVAR. Distinct advantages of EVAR over open repair have been demonstrated, including a less invasive operative exposure, decreased transfusion requirements, shortened intensive care unit and hospital stay, and decreased perioperative mortality. It is our opinion that in 2009, anatomically suitable patients should be offered EVAR as first-line therapy, except for the less common scenario of the young and fit patient, for which open repair should be strongly considered. Use of EVAR for ruptured AAAs also has shown promise, yielding survival results commensurate with the best single-center results with open repair for rupture. However, questions remain regarding the long-term efficacy of EVAR in preventing aneurysm-related death for all patients treated with this technique. As device improvements and technical advances continue, it is reasonable to expect that long-term results will improve as well. Furthermore, the advent of fenestrated and multi-branch endograft technology is expanding indications, and will continue to enlarge the percentage of patients who will be considered acceptable candidates for EVAR. Lastly, randomized clinical trials are under way to determine whether the generally accepted threshold of 5.5 cm for elective open AAA repair should be decreased in patients who are candidates for EVAR. Until further data emerge, standard guidelines for elective aneurysm repair should remain the norm. PMID- 19289027 TI - Optimal management of acute aortic dissection. AB - Acute aortic syndrome (AAS) encompasses a group of life-threatening aortic disorders that are increasing in prevalence. It classically presents with abrupt onset chest pain that is of maximum intensity at onset. The syndrome requires prompt recognition and efficient treatment to optimize outcome. Contrast-enhanced CT is most commonly used as the definitive diagnostic imaging modality. Patients must be expeditiously transferred to institutions with case experience and medical, surgical, and endovascular expertise. Stanford type A dissections involve the ascending aorta and require emergent surgical consideration, unless underlying comorbidities make surgical risk prohibitive. Patients with Stanford type B, or descending, aortic dissection are less prone to rupture, shock, and cardiac complications. These patients are initially medically managed with targeted blood pressure and heart rate control. Surgical management is reserved for patients with intractable or recurrent pain, aortic expansion, end-organ ischemia, or progression of dissection. The feasibility of endovascular treatment is established, and its role in management continues to expand. Long-term follow up with medical treatment and serial imaging of AAS patients is critical. Future directions in the diagnosis and treatment of AAS include using biomarkers to aid in diagnosis and prognosis, enhanced imaging with better resolution and reduced radiation exposure, and definition of the role of endovascular methods in acute and chronic settings. PMID- 19289028 TI - Unusual causes of intermittent claudication: popliteal artery entrapment syndrome, cystic adventitial disease, fibromuscular dysplasia, and endofibrosis. AB - In the general population, vascular causes of exercise-induced limb discomfort are most often the result of peripheral artery disease (PAD) due to atherosclerosis. However, several other clinical entities can often mimic the symptoms of atherosclerotic PAD of the lower extremities, particularly among younger patients with fewer risk factors for atherosclerosis, who often are more athletically fit than patients with PAD. Treatment for these entities often requires percutaneous or surgical intervention. This article reviews four uncommon vascular causes of exercise-induced limb discomfort: popliteal artery entrapment syndrome, cystic adventitial disease of the popliteal artery, fibromuscular dysplasia of the lower-extremity arteries, and endofibrosis of the iliac artery. PMID- 19289029 TI - Cervical artery dissection. AB - Spontaneous and traumatic cervical artery dissection is a common cause of stroke in the young. It generally carries an excellent prognosis if treatment is initiated early. Antiplatelet therapy may be as effective as or safer than warfarin, although no randomized prospective studies have addressed the issue of optimal medical therapy. Rarely, endovascular therapy may be indicated for the treatment of ruptured aneurysms or to prevent recurrent ischemia. PMID- 19289030 TI - Current management of thoracic outlet syndrome. AB - Thoracic outlet syndrome (TOS) is a condition caused by compression of the neurovascular structures leading to the arm passing through the thoracic outlet. There are three distinct types of TOS: neurogenic (95%), venous (4%-5%), and arterial (1%). Treatment algorithms depend on the type of TOS. Although statistically the most common type, neurogenic TOS can often be the most difficult to diagnose and treat. We have good follow-up data indicating that appropriately selected patients benefit from surgical intervention. Arterial and venous TOS often present more urgently with arterial or venous thrombosis. The thrombosis is typically recognized expeditiously by thorough history taking and physical examination, augmented by duplex ultrasonography. The restoration of blood flow, be it venous or arterial, often can be accomplished readily by thrombolysis. The key, however, comes in diagnosing the underlying structural component involved in the development of symptoms. To prevent recurrence, patients must undergo first rib resection and anterior scalenectomy, as well as resection of any rudimentary or cervical ribs. In the case of arterial TOS, the subclavian artery often requires reconstruction as well. Regardless of the type of TOS encountered, proper treatment requires a multidisciplinary approach. PMID- 19289031 TI - Rigidity-dependent cross talk between integrin and cadherin signaling. AB - Integrin-cadherin cross talk is an important aspect of cell function. We explored this signaling using substrates micropatterned with islands of fibronectin surrounded by E-cadherin, capturing the segregation of these signals in normal tissue. While MDCK cells were able to concurrently form adhesive structures with these two proteins, engagement of fibronectin by MCF-7 cells, an adenocarcinoma cell line, inhibited response of these cells to E-cadherin. We further demonstrated that this inhibition is rigidity dependent; on soft elastomer substrates with Young's modulus in the range of tens of kiloPascals, MCF-7 cells were able to engage both integrin and cadherin ligands. PMID- 19289032 TI - Relaxation dispersion NMR spectroscopy as a tool for detailed studies of protein folding. AB - Characterization of the mechanisms by which proteins fold into their native conformations is important not only for protein structure prediction and design but also because protein misfolding intermediates may play critical roles in fibril formation that are commonplace in neurodegenerative disorders. In practice, the study of folding pathways is complicated by the fact that for the most part intermediates are low-populated and short-lived so that biophysical studies are difficult. Due to recent methodological advances, relaxation dispersion NMR spectroscopy has emerged as a particularly powerful tool to obtain high-resolution structural information about protein folding events on the millisecond timescale. Applications of the methodology to study the folding of SH3 domains have shown that folding proceeds via previously undetected on-pathway intermediates, sometimes stabilized by nonnative long-range interactions. The relaxation dispersion approach provides a detailed kinetic and thermodynamic description of the folding process as well as the promise of obtaining an atomic level structural description of intermediate states. We review the concerted application of a variety of recently developed NMR relaxation dispersion experiments to obtain a "high-resolution" picture of the folding pathway of the A39V/N53P/V55L Fyn SH3 domain. PMID- 19289033 TI - Fluorescence correlation spectroscopy: the case of subdiffusion. AB - The theory of fluorescence correlation spectroscopy is revisited here for the case of subdiffusing molecules. Subdiffusion is assumed to stem from a continuous time random walk process with a fat-tailed distribution of waiting times and can therefore be formulated in terms of a fractional diffusion equation (FDE). The FDE plays the central role in developing the fluorescence correlation spectroscopy expressions, analogous to the role played by the simple diffusion equation for regular systems. Due to the nonstationary nature of the continuous time random walk/FDE, some interesting properties emerge that are amenable to experimental verification and may help in discriminating among subdiffusion mechanisms. In particular, the current approach predicts 1), a strong dependence of correlation functions on the initial time (aging); 2), sensitivity of correlation functions to the averaging procedure, ensemble versus time averaging (ergodicity breaking); and 3), that the basic mean-squared displacement observable depends on how the mean is taken. PMID- 19289034 TI - The tethered infinitesimal tori and spheres algorithm: a versatile calculator for axisymmetric problems in equilibrium membrane mechanics. AB - Constrained minimization of energy functionals is a central part, and usually the difficult part, of solving problems in the equilibrium mechanics of biological and biomimetic membranes. The inherent difficulties of the conventional variational-calculus approach prevents the numerical calculation involved from being made routine in the analyses of experimental results. We have developed a simulated annealing-based computational technique for routinizing the task of constrained minimization of energy functionals governing whole, or small patches of whole, fluid membranes with axisymmetry, spherical topology, and no domains of inhomogeneity. In this article, we describe the essential principles of the technique and apply it to five examples to demonstrate its versatility. It gives membrane shapes that are automatically stable to axisymmetric perturbations. Presently, it can account for constraints on 1), the membrane area or the effective membrane tension; 2), the enclosed volume or the effective pressure difference across the membrane thickness; and 3), the axial end-to-end distance or the applied axial point force. PMID- 19289035 TI - Computer modeling reveals that modifications of the histone tail charges define salt-dependent interaction of the nucleosome core particles. AB - Coarse-grained Langevin molecular dynamics computer simulations were conducted for systems that mimic solutions of nucleosome core particles (NCPs). The NCP was modeled as a negatively charged spherical particle representing the complex of DNA and the globular part of the histones combined with attached strings of connected charged beads modeling the histone tails. The size, charge, and distribution of the tails relative to the core were built to match real NCPs. Three models of NCPs were constructed to represent different extents of covalent modification on the histone tails: (nonmodified) recombinant (rNCP), acetylated (aNCP), and acetylated and phosphorylated (paNCP). The simulation cell contained 10 NCPs in a dielectric continuum with explicit mobile counterions and added salt. The NCP-NCP interaction is decisively dependent on the modification state of the histone tails and on salt conditions. Increasing the monovalent salt concentration (KCl) from salt-free to physiological concentration leads to NCP aggregation in solution for rNCP, whereas NCP associates are observed only occasionally in the system of aNCPs. In the presence of divalent salt (Mg(2+)), rNCPs form dense stable aggregates, whereas aNCPs form aggregates less frequently. Aggregates are formed via histone-tail bridging and accumulation of counterions in the regions of NCP-NCP contacts. The paNCPs do not show NCP-NCP interaction upon addition of KCl or in the presence of Mg(2+). Simulations for systems with a gradual substitution of K(+) for Mg(2+), to mimic the Mg(2+) titration of an NCP solution, were performed. The rNCP system showed stronger aggregation that occurred at lower concentrations of added Mg(2+), compared to the aNCP system. Additional molecular dynamics simulations performed with a single NCP in the simulation cell showed that detachment of the tails from the NCP core was modest under a wide range of salt concentrations. This implies that salt-induced tail dissociation of the histone tails from the globular NCP is not in itself a major factor in NCP-NCP aggregation. The approximation of coarse graining, with respect to the description of the NCP as a sphere with uniform charge distribution, was tested in control simulations. A more detailed description of the NCP did not change the main features of the results. Overall, the results of this work are in agreement with experimental data reported for NCP solutions and for chromatin arrays. PMID- 19289036 TI - Sensitivity of NFAT cycling to cytosolic calcium concentration: implications for hypertrophic signals in cardiac myocytes. AB - The nuclear factor of activated T-cell (NFAT) transcription factors play an important role in many biological processes, including pathological cardiac hypertrophy. Stimulated by calcium signals, NFAT is translocated to the nucleus where it can regulate hypertrophic genes (excitation-transcription coupling). In excitable cells, such as myocytes, calcium is a key second messenger for multiple signaling events, including excitation-contraction coupling. Whether the calcium signals due to excitation-contraction and excitation-transcription coupling coincide or how they can be differentiated is currently unclear. Here we construct a mathematical model of NFAT cycling fitted to skeletal myocyte and baby hamster kidney cell data. The model replicates key behavior with respect to sensitivity to calcineurin overexpression and to calcium oscillations. Finally, we measure the sensitivity of the system to a simulated hypertrophic calcium signal, against a background excitation-contraction coupling calcium oscillation. We find that NFAT cycling is sensitive to excitation-transcription coupling even when both calcium signals are in the same cellular compartment, thus showing that separation of the signals may not be necessary in vitro. PMID- 19289037 TI - Nitric oxide dynamics in truncated hemoglobin: docking sites, migration pathways, and vibrational spectroscopy from molecular dynamics simulations. AB - Atomistic simulations of nitric oxide (NO) dynamics and migration in the trHbN of Mycobacterium tuberculosis are reported. From extensive molecular dynamics simulations (48 ns in total), the structural and energetic properties of the ligand docking sites in the protein have been characterized and a connectivity network between the ligand docking sites has been built. Several novel migration and exit pathways are found and are analyzed in detail. The interplay between a hydrogen-bonding network involving residues Tyr(33) and Gln(58) and the bound O(2) ligand is discussed and the role of Phe(62) residue in ligand migration is examined. It is found that Phe(62) is directly involved in controlling ligand migration. This is reminiscent of His(64) in myoglobin, which also plays a central role in CO migration pathways. Finally, infrared spectra of the NO molecule in different ligand docking sites of the protein are calculated. The pocket-specific spectra are typically blue-shifted by 5-10 cm(-1), which should be detectable in future spectroscopic experiments. PMID- 19289038 TI - Protein structure prediction by pro-Sp3-TASSER. AB - An automated protein structure prediction algorithm, pro-sp3 Threading/ASSEmbly/Refinement (TASSER), is described and benchmarked. Structural templates are identified using five different scoring functions derived from the previously developed threading methods PROSPECTOR_3 and SP(3). Top templates identified by each scoring function are combined to derive contact and distant restraints for subsequent model refinement by short TASSER simulations. For Medium/Hard targets (those with moderate to poor quality templates and/or alignments), alternative template alignments are also generated by parametric alignment and the top models selected by TASSER-QA are included in the contact and distance restraint derivation. Then, multiple short TASSER simulations are used to generate an ensemble of full-length models. Subsequently, the top models are selected from the ensemble by TASSER-QA and used to derive TASSER contacts and distant restraints for another round of full TASSER refinement. The final models are selected from both rounds of TASSER simulations by TASSER-QA. We compare pro-sp3-TASSER with our previously developed MetaTASSER method (enhanced with chunk-TASSER for Medium/Hard targets) on a representative test data set of 723 proteins <250 residues in length. For the 348 proteins classified as easy targets (those templates with good alignments and global structure similarity to the target), the cumulative TM-score of the best of top five models by pro-sp3 TASSER shows a 2.1% improvement over MetaTASSER. For the 155/220 medium/hard targets, the improvements in TM-score are 2.8% and 2.2%, respectively. All improvements are statistically significant. More importantly, the number of foldable targets (those having models whose TM-score to native >0.4 in the top five clusters) increases from 472 to 497 for all targets, and the relative increases for medium and hard targets are 10% and 15%, respectively. A server that implements the above algorithm is available at http://cssb.biology.gatech.edu/skolnick/webservice/pro-sp3-TASSER/. The source code is also available upon request. PMID- 19289039 TI - Coupling between normal modes drives protein conformational dynamics: illustrations using allosteric transitions in myosin II. AB - Structure-based elastic network models (ENMs) have been remarkably successful in describing conformational transitions in a variety of biological systems. Low frequency normal modes are usually calculated from the ENM that characterizes elastic interactions between residues in contact in a given protein structure with a uniform force constant. To explore the dynamical effects of nonuniform elastic interactions, we calculate the robustness and coupling of the low frequency modes in the presence of nonuniform variations in the ENM force constant. The variations in the elastic interactions, approximated here by Gaussian noise, approximately account for perturbation effects of heterogeneous residue-residue interactions or evolutionary sequence changes within a protein family. First-order perturbation theory provides an efficient and qualitatively correct estimate of the mode robustness and mode coupling for finite perturbations to the ENM force constant. The mode coupling analysis and the mode robustness analysis identify groups of strongly coupled modes that encode for protein functional motions. We illustrate the new concepts using myosin II motor protein as an example. The biological implications of mode coupling in tuning the allosteric couplings among the actin-binding site, the nucleotide-binding site, and the force-generating converter and lever arm in myosin isoforms are discussed. We evaluate the robustness of the correlation functions that quantify the allosteric couplings among these three key structural motifs. PMID- 19289040 TI - Ion selectivity in the KcsA potassium channel from the perspective of the ion binding site. AB - To understand the thermodynamic exclusion of Na(+) relative to K(+) from the S(2) site of the selectivity filter, the distribution P(X)(epsilon) (X = K(+) or Na(+)) of the binding energy (epsilon) of the ion with the channel is analyzed using the potential distribution theorem. By expressing the excess chemical potential of the ion as a sum of mean-field epsilon and fluctuation mu(flux,X)(ex) contributions, we find that selectivity arises from a higher value of mu(flux,Na(+))(ex) relative to mu(flux,K(+))(ex). To understand the role of site-site interactions on mu(ex)(flux,X), we decompose P(X)(epsilon) into n dependent distributions, where n is the number of ion-coordinating ligands within a distance lambda from the ion. For lambda comparable to typical ion-oxygen bond distances, investigations building on this multistate model reveal an inverse correlation between favorable ion-site and site-site interactions: the ion coordination states that most influence the thermodynamics of the ion are also those for which the binding site is energetically less strained and vice versa. This correlation motivates understanding entropic effects in ion binding to the site and leads to the finding that mu(flux,X)(ex) is directly proportional to the average site-site interaction energy, a quantity that is sensitive to the chemical type of the ligand coordinating the ion. Increasing the coordination number around Na(+) only partially accounts for the observed magnitude of selectivity; acknowledging the chemical type of the ion-coordinating ligand is essential. PMID- 19289041 TI - Nonequilibrium self-assembly of a filament coupled to ATP/GTP hydrolysis. AB - We study the stochastic dynamics of growth and shrinkage of single actin filaments or microtubules taking into account insertion, removal, and ATP/GTP hydrolysis of subunits. The resulting phase diagram contains three different phases: two phases of unbounded growth: a rapidly growing phase and an intermediate phase, and one bounded growth phase. We analyze all these phases, with an emphasis on the bounded growth phase. We also discuss how hydrolysis affects force-velocity curves. The bounded growth phase shows features of dynamic instability, which we characterize in terms of the time needed for the ATP/GTP cap to disappear as well as the time needed for the filament to reach a length of zero (i.e., to collapse) for the first time. We obtain exact expressions for all these quantities, which we test using Monte Carlo simulations. PMID- 19289042 TI - A theoretical model for the mechanical unfolding of repeat proteins. AB - We consider the mechanical stretching of a polypeptide chain formed by multiple interacting repeats. The folding thermodynamics and the interactions among the repeats are described by the Ising model. Unfolded repeats act as soft entropic springs, whereas folded repeats respond to a force as stiffer springs. We show that the resulting force-extension curve may exhibit a pronounced force maximum corresponding to the unfolding of the first repeat. This event is followed by the unfolding of the remaining repeats, which takes place at a lower force. As the protein extension is increased, the force-extension curve of a sufficiently long repeat protein displays a plateau, where the force remains nearly constant and the protein unfolds sequentially so that the number of unfolded repeats is proportional to the extension. Such a sequential mechanical unfolding mechanism is displayed even by the repeat proteins whose thermal denaturation is highly cooperative, provided that they are long enough. By contrast, the unfolding of short repeat progressions can be cooperative. PMID- 19289043 TI - Thermodynamic pathways to genome spatial organization in the cell nucleus. AB - The architecture of the eukaryotic genome is characterized by a high degree of spatial organization. Chromosomes occupy preferred territories correlated to their state of activity and, yet, displace their genes to interact with remote sites in complex patterns requiring the orchestration of a huge number of DNA loci and molecular regulators. Far from random, this organization serves crucial functional purposes, but its governing principles remain elusive. By computer simulations of a statistical mechanics model, we show how architectural patterns spontaneously arise from the physical interaction between soluble binding molecules and chromosomes via collective thermodynamics mechanisms. Chromosomes colocalize, loops and territories form, and find their relative positions as stable thermodynamic states. These are selected by thermodynamic switches, which are regulated by concentrations/affinity of soluble mediators and by number/location of their attachment sites along chromosomes. Our thermodynamic switch model of nuclear architecture, thus, explains on quantitative grounds how well-known cell strategies of upregulation of DNA binding proteins or modification of chromatin structure can dynamically shape the organization of the nucleus. PMID- 19289044 TI - Modeling effects of human single nucleotide polymorphisms on protein-protein interactions. AB - A large set of three-dimensional structures of 264 protein-protein complexes with known nonsynonymous single nucleotide polymorphisms (nsSNPs) at the interface was built using homology-based methods. The nsSNPs were mapped on the proteins' structures and their effect on the binding energy was investigated with CHARMM force field and continuum electrostatic calculations. Two sets of nsSNPs were studied: disease annotated Online Mendelian Inheritance in Man (OMIM) and nonannotated (non-OMIM). It was demonstrated that OMIM nsSNPs tend to destabilize the electrostatic component of the binding energy, in contrast with the effect of non-OMIM nsSNPs. In addition, it was shown that the change of the binding energy upon amino acid substitutions is not related to the conservation of the net charge, hydrophobicity, or hydrogen bond network at the interface. The results indicate that, generally, the effect of nsSNPs on protein-protein interactions cannot be predicted from amino acids' physico-chemical properties alone, since in many cases a substitution of a particular residue with another amino acid having completely different polarity or hydrophobicity had little effect on the binding energy. Analysis of sequence conservation showed that nsSNP at highly conserved positions resulted in a large variance of the binding energy changes. In contrast, amino acid substitutions corresponding to nsSNPs at nonconserved positions, on average, were not found to have a large effect on binding affinity. pKa calculations were performed and showed that amino acid substitutions could change the wild-type proton uptake/release and thus resulting in different pH dependence of the binding energy. PMID- 19289045 TI - The origin of short transcriptional pauses. AB - RNA polymerases are protein molecular machines that transcribe genetic information from DNA into RNA. The elongation of the RNA molecule is frequently interrupted by pauses, the detailed nature of which remains controversial. Here we ask whether backtracking, the central mechanism behind long pauses, could also be responsible for short pauses normally attributed to the ubiquitous pause state. To this end, we model backtracking as a force-biased random walk, giving rise to a broad distribution of pause durations as observed in experiments. Importantly, we find that this single mechanism naturally generates two populations of pauses that are distinct both in duration and trajectory: long time pauses with the expected behavior of diffusive backtracks, and a new class of short-time backtracks with characteristics similar to those of the ubiquitous pause. These characteristics include an apparent force insensitivity and immobility of the polymerase. Based on these results and a quantitative comparison to published pause trajectories measured with optical tweezers, we suggest that a significant fraction of short pauses are simply due to backtracking. PMID- 19289046 TI - Structure and membrane interactions of the antibiotic peptide dermadistinctin K by multidimensional solution and oriented 15N and 31P solid-state NMR spectroscopy. AB - DD K, a peptide first isolated from the skin secretion of the Phyllomedusa distincta frog, has been prepared by solid-phase chemical peptide synthesis and its conformation was studied in trifluoroethanol/water as well as in the presence of sodium dodecyl sulfate and dodecylphosphocholine micelles or small unilamellar vesicles. Multidimensional solution NMR spectroscopy indicates an alpha-helical conformation in membrane environments starting at residue 7 and extending to the C-terminal carboxyamide. Furthermore, DD K has been labeled with (15)N at a single alanine position that is located within the helical core region of the sequence. When reconstituted into oriented phosphatidylcholine membranes the resulting (15)N solid-state NMR spectrum shows a well-defined helix alignment parallel to the membrane surface in excellent agreement with the amphipathic character of DD K. Proton-decoupled (31)P solid-state NMR spectroscopy indicates that the peptide creates a high level of disorder at the level of the phospholipid headgroup suggesting that DD K partitions into the bilayer where it severely disrupts membrane packing. PMID- 19289047 TI - Structure of ceramide-1-phosphate at the air-water solution interface in the absence and presence of Ca2+. AB - Ceramide-1-phosphate, the phosphorylated form of ceramide, gained attention recently due to its diverse intracellular roles, in particular in inflammation mediated by cPLA(2)alpha. However, surprisingly little is known about the physical chemical properties of this lipid and its potential impact on physiological function. For example, the presence of Ca(2+) is indispensable for the interaction of Cer-1-P with the C2 domain of cPLA(2)alpha. We report on the structure and morphology of Cer-1-P in monomolecular layers at the air/water solution interface in the absence and presence of Ca(2+) using diverse biophysical techniques, including synchrotron x-ray reflectivity and grazing angle diffraction, to gain insight into the role and function of Cer-1-P in biomembranes. We show that relatively small changes in pH and the presence of monovalent cations dramatically affect the behavior of Cer-1-P. On pure water Cer 1-P forms a solid monolayer despite the negative charge of the phosphomonoester headgroup. In contrast, pH 7.2 buffer yields a considerably less solid-like monolayer, indicating that charge-charge repulsion becomes important at higher pH. Calcium was found to bind strongly to the headgroup of Cer-1-P even in the presence of a 100-fold larger Na(+) concentration. Analysis of the x-ray reflectivity data allowed us to estimate how much Ca(2+) is bound to the headgroup, approximately 0.5 Ca(2+) and approximately 1.0 Ca(2+) ions per Cer-1-P molecule for the water and buffer subphase respectively. These results can be qualitatively understood based on the molecular structure of Cer-1-P and the electrostatic/hydrogen-bond interactions of its phosphomonoester headgroup. Biological implications of our results are also discussed. PMID- 19289048 TI - Ceramide-1-phosphate, in contrast to ceramide, is not segregated into lateral lipid domains in phosphatidylcholine bilayers. AB - Sphingolipids are key lipid regulators of cell viability: ceramide is one of the key molecules in inducing programmed cell death (apoptosis), whereas other sphingolipids, such as ceramide 1-phosphate, are mitogenic. The thermotropic and structural behavior of binary systems of N-hexadecanoyl-D-erythro-ceramide (C(16) ceramide) or N-hexadecanoyl-D-erythro-ceramide-1-phosphate (C(16)-ceramide-1 phosphate; C(16)-C1P) with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was studied with DSC and deuterium nuclear magnetic resonance ((2)H-NMR). Partial phase diagrams (up to a mole fraction of sphingolipids X = 0.40) for both mixtures were constructed based on DSC and (2)H-NMR observations. For C(16) ceramide-containing bilayers DSC heating scans showed already at X(cer) = 0.025 a complex structure of the main-phase transition peak suggestive of lateral-phase separation. The transition width increased significantly upon increasing X(cer), and the upper-phase boundary temperature of the mixture shifted to approximately 65 degrees C at X(cer) = 0.40. The temperature range over which (2)H-NMR spectra of C(16)-ceramide/DPPC-d(62) mixtures displayed coexistence of gel and liquid crystalline domains increased from approximately 10 degrees for X(cer) = 0.1 to approximately 21 degrees for X(cer) = 0.4. For C16-C1P/DPPC mixtures, DSC and (2)H-NMR observations indicated that two-phase coexistence was limited to significantly narrower temperature ranges for corresponding C1P concentrations. To complement these findings, C(16)-ceramide/1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC) and C16-C1P/POPC mixtures were also studied by (2)H-NMR and fluorescence techniques. These observations indicate that DPPC and POPC bilayers are significantly less perturbed by C(16)-C1P than by C(16)-ceramide and that C(16)-C1P is miscible within DPPC bilayers at least up to X(C1P) = 0.30. PMID- 19289049 TI - Development of a CP 31P NMR broadline simulation methodology for studying the interactions of antihypertensive AT1 antagonist losartan with phospholipid bilayers. AB - A cross-polarization (CP) (31)P NMR broadline simulation methodology was developed for studying the effects of drugs in phospholipids bilayers. Based on seven-parameter fittings, this methodology provided information concerning the conformational changes and dynamics effects of losartan in the polar region of the dipalmitoylphosphatidylcholine bilayers. The test molecule for this study was losartan, an antihypertensive drug known to exert its effect on AT(1) transmembrane receptors. The results were complemented and compared with those of differential scanning calorimetry, solid-state (13)C NMR spectroscopy, Raman spectroscopy, and electron spin resonance. More specifically, these physical chemical methodologies indicated that the amphipathic losartan molecule interacts with the hydrophilic-head zone of the lipid bilayers. The CP (31)P NMR broadline simulations showed that the lipid molecules in the bilayers containing losartan displayed greater collective tilt compared to the tilt displayed by the load-free bilayers, indicating improved packing. The Raman results displayed a decrease in the trans/gauche ratio and increased intermolecular interactions of the acyl chains in the liquid crystalline phase. Additional evidence, suggesting that losartan possibly anchors in the realm of the headgroup, was derived from upfield shift of the average chemical shift sigma(iso) of the (31)P signal in the presence of losartan and from shift of the observed peak at 715 cm(-1) attributed to C-N stretching in the Raman spectra. PMID- 19289050 TI - Some cardiomyopathy-causing troponin I mutations stabilize a functional intermediate actin state. AB - We examined four cardiomyopathy-causing mutations of troponin I that appear to disturb function by altering the distribution of thin filament states. The R193H (mouse) troponin I mutant had greater than normal actin-activated myosin-S1 ATPase activity in both the presence and absence of calcium. The rate of ATPase activity was the same as that of the wild-type at near-saturating concentrations of the activator, N-ethylmaleimide-S1. This mutant appeared to function by stabilizing the active state of thin filaments. Mutations D191H, R146G, and R146W had lower ATPase activities in the presence of calcium, but higher activities in the absence of calcium. These effects were most pronounced with mutations at position 146. For all three mutants the rates were similar to those of the wild type at near-saturating concentrations of N-ethylmaleimide-S1. These results, combined with previous results, show that any alteration in the normal distribution of actomyosin states is capable of producing cardiomyopathy. The results of the D191H, R146G, and R146W mutations are most readily explained if the intermediate state of regulated actin has a unique function. The intermediate state appears to have an ability to accelerate the rate of ATP hydrolysis by myosin that exceeds that of the inactive state. PMID- 19289051 TI - DNA sequence-directed organization of chromatin: structure-based computational analysis of nucleosome-binding sequences. AB - The folding of DNA on the nucleosome core particle governs many fundamental issues in eukaryotic molecular biology. In this study, an updated set of sequence dependent empirical "energy" functions, derived from the structures of other protein-bound DNA molecules, is used to investigate the extent to which the architecture of nucleosomal DNA is dictated by its underlying sequence. The potentials are used to estimate the cost of deforming a collection of sequences known to bind or resist uptake in nucleosomes along various left-handed superhelical pathways and to deduce the features of sequence contributing to a particular structural form. The deformation scores reflect the choice of template, the deviations of structural parameters at each step of the nucleosome bound DNA from their intrinsic values, and the sequence-dependent "deformability" of a given dimer. The correspondence between the computed scores and binding propensities points to a subtle interplay between DNA sequence and nucleosomal folding, e.g., sequences with periodically spaced pyrimidine-purine steps deform at low cost along a kinked template whereas sequences that resist deformation prefer a smoother spatial pathway. Successful prediction of the known settings of some of the best-resolved nucleosome-positioning sequences, however, requires a template with "kink-and-slide" steps like those found in high-resolution nucleosome structures. PMID- 19289052 TI - A mechanism of energy dissipation in cyanobacteria. AB - When grown under a variety of stress conditions, cyanobacteria express the isiA gene, which encodes the IsiA pigment-protein complex. Overexpression of the isiA gene under iron-depletion stress conditions leads to the formation of large IsiA aggregates, which display remarkably short fluorescence lifetimes and thus a strong capacity to dissipate energy. In this work we investigate the underlying molecular mechanism responsible for chlorophyll fluorescence quenching. Femtosecond transient absorption spectroscopy allowed us to follow the process of energy dissipation in real time. The light energy harvested by chlorophyll pigments migrated within the system and eventually reaches a quenching site where the energy is transferred to a carotenoid-excited state, which dissipates it by decaying to the ground state. We compare these findings with those obtained for the main light-harvesting complex in green plants (light-harvesting complex II) and artificial light-harvesting antennas, and conclude that all of these systems show the same mechanism of energy dissipation, i.e., one or more carotenoids act as energy dissipators by accepting energy via low-lying singlet-excited S(1) states and dissipating it as heat. PMID- 19289053 TI - Femtosecond carotenoid to retinal energy transfer in xanthorhodopsin. AB - Xanthorhodopsin of the extremely halophilic bacterium Salinibacter ruber represents a novel antenna system. It consists of a carbonyl carotenoid, salinixanthin, bound to a retinal protein that serves as a light-driven transmembrane proton pump similar to bacteriorhodopsin of archaea. Here we apply the femtosecond transient absorption technique to reveal the excited-state dynamics of salinixanthin both in solution and in xanthorhodopsin. The results not only disclose extremely fast energy transfer rates and pathways, they also reveal effects of the binding site on the excited-state properties of the carotenoid. We compared the excited-state dynamics of salinixanthin in xanthorhodopsin and in NaBH(4)-treated xanthorhodopsin. The NaBH(4) treatment prevents energy transfer without perturbing the carotenoid binding site, and allows observation of changes in salinixanthin excited-state dynamics related to specific binding. The S(1) lifetimes of salinixanthin in untreated and NaBH(4) treated xanthorhodopsin were identical (3 ps), confirming the absence of the S(1) mediated energy transfer. The kinetics of salinixanthin S(2) decay probed in the near-infrared region demonstrated a change of the S(2) lifetime from 66 fs in untreated xanthorhodopsin to 110 fs in the NaBH(4)-treated protein. This corresponds to a salinixanthin-retinal energy transfer time of 165 fs and an efficiency of 40%. In addition, binding of salinixanthin to xanthorhodopsin increases the population of the S(*) state that decays in 6 ps predominantly to the ground state, but a small fraction (<10%) of the S(*) state generates a triplet state. PMID- 19289054 TI - Molecular dynamics simulations reveal that Tyr-317 phosphorylation reduces Shc binding affinity for phosphotyrosyl residues of epidermal growth factor receptor. AB - The Src homology 2 (SH2) and collagen domain protein Shc plays a pivotal role in signaling via tyrosine kinase receptors, including epidermal growth factor receptor (EGFR). Shc binding to phospho-tyrosine residues on activated receptors is mediated by the SH2 and phospho-tyrosine binding (PTB) domains. Subsequent phosphorylation on Tyr-317 within the Shc linker region induces Shc interactions with Grb2-Son of Sevenless that initiate Ras-mitogen-activated protein kinase signaling. We use molecular dynamics simulations of full-length Shc to examine how Tyr-317 phosphorylation controls Shc conformation and interactions with EGFR. Our simulations reveal that Shc tyrosine phosphorylation results in a significant rearrangement of the relative position of its domains, suggesting a key conformational change. Importantly, computational estimations of binding affinities show that EGFR-derived phosphotyrosyl peptides bind with significantly more strength to unphosphorylated than to phosphorylated Shc. Our results unveil what we believe is a novel structural phenomenon, i.e., tyrosine phosphorylation of Shc within its linker region regulates the binding affinity of SH2 and PTB domains for phosphorylated Shc partners, with important implications for signaling dynamics. PMID- 19289055 TI - Agitation and high ionic strength induce amyloidogenesis of a folded PDZ domain in native conditions. AB - Amyloid fibril formation is a distinctive hallmark of a number of degenerative diseases. In this process, protein monomers self-assemble to form insoluble structures that are generally referred to as amyloid fibrils. We have induced in vitro amyloid fibril formation of a PDZ domain by combining mechanical agitation and high ionic strength under conditions otherwise close to physiological (pH 7.0, 37 degrees C, no added denaturants). The resulting aggregates enhance the fluorescence of the thioflavin T dye via a sigmoidal kinetic profile. Both infrared spectroscopy and circular dichroism spectroscopy detect the formation of a largely intermolecular beta-sheet structure. Atomic force microscopy shows straight, rod-like fibrils that are similar in appearance and height to mature amyloid-like fibrils. Under these conditions, before aggregation, the protein domain adopts an essentially native-like structure and an even higher conformational stability (DeltaG(U-F)(H2O)). These results show a new method for converting initially folded proteins into amyloid-like aggregates. The methodological approach used here does not require denaturing conditions; rather, it couples agitation with a high ionic strength. Such an approach offers new opportunities to investigate protein aggregation under conditions in which a globular protein is initially folded, and to elucidate the physical forces that promote amyloid fibril formation. PMID- 19289056 TI - The effect of loops on the structural organization of alpha-helical membrane proteins. AB - Loops connecting the transmembrane (TM) alpha-helices in membrane proteins are expected to affect the structural organization of the thereby connected helices and the helical bundles as a whole. This effect, which has been largely ignored previously, is studied here by analyzing the x-ray structures of 41 alpha-helical membrane proteins. First we define the loop flexibility ratio, R, and find that 53% of the loops are stretched, where a stretched loop constrains the distance between the two connected helices. The significance of this constraining effect is supported by experiments carried out with bacteriorhodopsin and rhodopsin, in which cutting or eliminating their (predominately stretched) loops has led to a decrease in protein stability, and for rhodopsin, in most cases, also to the destruction of the structure. We show that for nonstretched loops in the extramembranous regions, the fraction of hydrophobic residues is comparable to that for soluble proteins; furthermore (as is also the case for soluble proteins), the hydrophobic residues in these regions are preferentially buried. This is expected to lead to the compact structural organization of the loops, which is transferred to the TM helices, causing them to assemble. We argue that a soluble protein complexed with a membrane protein similarly promotes compactness; other properties of such complexes are also studied. We calculate complementary attractive interactions between helices, including hydrogen bonds and van der Waals interactions of sequential motifs, such as GXXXG. The relative and combined effects of all these factors on the association of the TM helices are discussed and protein structures with only a few of these factors are analyzed. Our study emphasizes the need for classifying membrane proteins into groups according to structural organization. This classification should be considered when procedures for structural analysis or prediction are developed and applied. Detailed analysis of each structure is provided at http://flan.blm.cs.cmu.edu/memloop/ PMID- 19289057 TI - Fluid shear induces conformation change in human blood protein von Willebrand factor in solution. AB - Many of the physiological functions of von Willebrand Factor (VWF), including its binding interaction with blood platelets, are regulated by the magnitude of applied fluid/hydrodynamic stress. We applied two complementary strategies to study the effect of fluid forces on the solution structure of VWF. First, small angle neutron scattering was used to measure protein conformation changes in response to laminar shear rates (G) up to 3000/s. Here, purified VWF was sheared in a quartz Couette cell and protein conformation was measured in real time over length scales from 2-140 nm. Second, changes in VWF structure up to 9600/s were quantified by measuring the binding of a fluorescent probe 1,1'-bis(anilino)-4 ,4'-bis(naphthalene)-8,8'-disulfonate (bis-ANS) to hydrophobic pockets exposed in the sheared protein. Small angle neutron scattering studies, coupled with quantitative modeling, showed that VWF undergoes structural changes at G < 3000/s. These changes were most prominent at length scales <10 nm (scattering vector (q) range >0.6/nm). A mathematical model attributes these changes to the rearrangement of domain level features within the globular section of the protein. Studies with bis-ANS demonstrated marked increase in bis-ANS binding at G > 2300/s. Together, the data suggest that local rearrangements at the domain level may precede changes at larger-length scales that accompany exposure of protein hydrophobic pockets. Changes in VWF conformation reported here likely regulate protein function in response to fluid shear. PMID- 19289058 TI - Atomistic insights into regulatory mechanisms of the HER2 tyrosine kinase domain: a molecular dynamics study. AB - HER2 (ErbB2/Neu) is a receptor tyrosine kinase belonging to the epidermal growth factor receptor (EGFR)/ErbB family and is overexpressed in 20-30% of human breast cancers. Although several crystal structures of ErbB kinases have been solved, the precise mechanism of HER2 activation remains unknown, and it has been suggested that HER2 is unique in its requirement for phosphorylation of Y877, a key tyrosine residue located in the activation loop. To elucidate mechanistic details of kinase domain regulation, we performed molecular dynamics simulations of a homology-modeled HER2 kinase structure in active and inactive conformations. Principal component analysis of the atomistic fluctuations reveals a tight coupling between the activation loop and catalytic loop that may contribute to alignment of residues required for catalysis in the active kinase. The free energy perturbation method is also employed to predict a role for phosphorylated Y877 in stabilizing the kinase conformations. Finally, simulation results are presented for a HER2/EGFR heterodimer and reveal that the dimeric interface induces a rearrangement of the alphaC helix toward the active conformation. Elucidation of the molecular regulatory mechanisms in HER2 will help establish structure-function relationships in the wild-type kinase, as well as predict mutations with a propensity for constitutive activation in HER2-mediated cancers. PMID- 19289059 TI - The effects of ADF/cofilin and profilin on the conformation of the ATP-binding cleft of monomeric actin. AB - Actin depolymerizing factor (ADF)/cofilin and profilin are small actin-binding proteins, which have central roles in cytoskeletal dynamics in all eukaryotes. When bound to an actin monomer, ADF/cofilins inhibit the nucleotide exchange, whereas most profilins accelerate the nucleotide exchange on actin monomers. In this study the effects of ADF/cofilin and profilin on the accessibility of the actin monomer's ATP-binding pocket was investigated by a fluorescence spectroscopic method. The fluorescence of the actin bound epsilon-ATP was quenched with a neutral quencher (acrylamide) in steady-state and time dependent experiments, and the data were analyzed with a complex form of the Stern-Volmer equation. The experiments revealed that in the presence of ADF/cofilin the accessibility of the bound epsilon-ATP decreased, indicating a closed and more compact ATP-binding pocket induced by the binding of ADF/cofilin. In the presence of profilin the accessibility of the bound epsilon-ATP increased, indicating a more open and approachable protein matrix around the ATP-binding pocket. The results of the fluorescence quenching experiments support a structural mechanism regarding the regulation of the nucleotide exchange on actin monomers by ADF/cofilin and profilin. PMID- 19289060 TI - Elastic energy driven polymerization. AB - We present a molecular system where polymerization is controlled externally by tuning the elastic energy of the monomers. The elastic energy, provided by a DNA molecular spring, destabilizes the monomer state through a process analogous to domain swapping. This energy can be large (of approximately 10 kT) and thus drive polymerization at relatively low monomer concentrations. The monomer-dimer equilibrium provides a measurement of the elastic energy of the monomer, which in this construction appears limited by kink formation in the DNA molecular spring, in accord with previous theoretical and experimental investigations of the elasticity of sharply bent DNA. PMID- 19289061 TI - Existence of different structural intermediates on the fibrillation pathway of human serum albumin. AB - The fibrillation propensity of the multidomain protein human serum albumin (HSA) was analyzed under different solution conditions. The aggregation kinetics, protein conformational changes upon self-assembly, and structure of the different intermediates on the fibrillation pathway were determined by means of thioflavin T (ThT) fluorescence and Congo Red absorbance; far- and near-ultraviolet circular dichroism; tryptophan fluorescence; Fourier transform infrared spectroscopy; x ray diffraction; and transmission electron, scanning electron, atomic force, and microscopies. HSA fibrillation extends over several days of incubation without the presence of a lag phase, except for HSA samples incubated at acidic pH and room temperature in the absence of electrolyte. The absence of a lag phase occurs if the initial aggregation is a downhill process that does not require a highly organized and unstable nucleus. The fibrillation process is accompanied by a progressive increase in the beta-sheet (up to 26%) and unordered conformation at the expense of alpha-helical conformation, as revealed by ThT fluorescence and circular dichroism and Fourier transform infrared spectroscopies, but changes in the secondary structure contents depend on solution conditions. These changes also involve the presence of different structural intermediates in the aggregation pathway, such as oligomeric clusters (globules), bead-like structures, and ring-shaped aggregates. We suggest that fibril formation may take place through the role of association-competent oligomeric intermediates, resulting in a kinetic pathway via clustering of these oligomeric species to yield protofibrils and then fibrils. The resultant fibrils are elongated but curly, and differ in length depending on solution conditions. Under acidic conditions, circular fibrils are commonly observed if the fibrils are sufficiently flexible and long enough for the ends to find themselves regularly in close proximity to each other. These fibrils can be formed by an antiparallel arrangement of beta-strands forming the beta-sheet structure of the HSA fibrils as the most probable configuration. Very long incubation times lead to a more complex morphological variability of amyloid mature fibrils (i.e., long straight fibrils, flat-ribbon structures, laterally connected fibers, etc.). We also observed that mature straight fibrils can also grow by protein oligomers tending to align within the immediate vicinity of the fibers. This filament + monomers/oligomers scenario is an alternative pathway to the otherwise dominant filament + filament manner of the protein fibril's lateral growth. Conformational preferences for a certain pathway to become active may exist, and the influence of environmental conditions such as pH, temperature, and salt must be considered. PMID- 19289062 TI - Mitigating unwanted photophysical processes for improved single-molecule fluorescence imaging. AB - Organic fluorophores common to fluorescence-based investigations suffer from unwanted photophysical properties, including blinking and photobleaching, which limit their overall experimental performance. Methods to control such processes are particularly important for single-molecule fluorescence and fluorescence resonance energy transfer imaging where uninterrupted, stable fluorescence is paramount. Fluorescence and FRET-based assays have been carried out on dye labeled DNA and RNA-based systems to quantify the effect of including small molecule solution additives on the fluorescence and FRET behaviors of both cyanine and Alexa fluorophores. A detailed dwell time analysis of the fluorescence and FRET trajectories of more than 200,000 individual molecules showed that two compounds identified previously as triplet state quenchers, cyclooctatetraene, and Trolox, as well as 4-nitrobenzyl alcohol, act to favorably attenuate blinking, photobleaching, and influence the rate of photoresurrection in a concentration-dependent and context-dependent manner. In both biochemical systems examined, a unique cocktail of compounds was shown to be optimal for imaging performance. By simultaneously providing the most rapid and direct access to multiple photophysical kinetic parameters, smFRET imaging provides a powerful avenue for future investigations aimed at discovering new compounds, and effective combinations thereof. These efforts may ultimately facilitate tuning organic dye molecule performance according to each specific experimental demand. PMID- 19289063 TI - Extracellular space volume measured by two-color pulsed dye infusion with microfiberoptic fluorescence photodetection. AB - The extracellular space (ECS) is the aqueous matrix surrounding cells in solid tissues. The only method to measure ECS volume fraction (alpha) in vivo has been tetramethylammonium iontophoresis, a technically challenging method developed more than 25 years ago. We report a simple, quantitative method to measure alpha by microfiberoptic fluorescence detection of a self-quenched green dye, calcein, and a reference red dye, sulforhodamine 101, after pulsed iontophoretic infusion. The idea is that the maximum increase in calcein fluorescence after iontophoresis is proportional to the aqueous volume into which the dye is deposited. We validated the method theoretically, and experimentally, using cell-embedded gels with specified alpha and ECS viscosity. Measurements in living mice gave alpha of 0.20 +/- 0.01 in brain, 0.13 +/- 0.02 in kidney and 0.074 +/- 0.01 in skeletal muscle. The technical simplicity of the "pulsed-infusion microfiberoptic photodetection" method developed here should allow elucidation of the relatively understudied biological roles of the ECS. PMID- 19289064 TI - Fluorescence fluctuation spectroscopy of mCherry in living cells. AB - The red fluorescent protein mCherry is of considerable interest for fluorescence fluctuation spectroscopy (FFS), because the wide separation in color between mCherry and green fluorescent protein provides excellent conditions for identifying protein interactions inside cells. This two-photon study reveals that mCherry exists in more than a single brightness state. Unbiased analysis of the data needs to account for the presence of multiple states. We introduce a two state model that successfully describes the brightness and fluctuation amplitude of mCherry. The properties of the two states are characterized by FFS and fluorescence lifetime experiments. No interconversion between the two states was observed over the experimentally probed timescales. The effect of fluorescence resonance energy transfer between enhanced green fluorescent protein (EGFP) and mCherry is incorporated into the two-state model to describe protein hetero oligomerization. The model is verified by comparing the predicted and measured brightness and fluctuation amplitude of several fusion proteins that contain mCherry and EGFP. In addition, hetero-fluorescence resonance energy transfer between mCherry molecules in different states is detected, but its influence on FFS parameters is small enough to be negligible. Finally, the two-state model is applied to study protein oligomerization in living cells. We demonstrate that the model successfully describes the homodimerization of nuclear receptors. In addition, we resolved a mixture of interacting and noninteracting proteins labeled with EGFP and mCherry. These results provide the foundation for quantitative applications of mCherry in FFS studies. PMID- 19289065 TI - Quantifying cell-surface biomarker expression in thick tissues with ratiometric three-dimensional microscopy. AB - The burgeoning fields of in vivo three-dimensional (3D) microscopy and endomicroscopy, as well as ex vivo tissue cytometry have introduced new challenges for tissue preparation and staining with exogenous molecular contrast agents. These challenges include effective delivery of the agents, and once delivered, distinguishing between bound verses unbound molecular probes. If applied topically, there are additional issues with rinsing off unbound probe, which can be nonuniform and inefficient in thick tissues, thus leading to ambiguous contrast and a large nonspecific background that may obscure molecule specific staining. Therefore, we have developed a ratiometric 3D microscopy scheme that not only reduces the effects of nonspecific sources of contrast, but also enables quantification of the relative binding affinity of imaging probes to their biomarker targets. Here we demonstrate this ratiometric approach by simultaneously imaging a HER2/neu (erbB2)-targeted monoclonal antibody labeled with one fluorophore and an isotype-matched negative control antibody labeled with another fluorophore. By taking a pixel-by-pixel calibrated ratio between the signals from each fluorescent image channel, accurate quantification of specific versus nonspecific binding affinity is achieved with cultured cells, yielding data that are in agreement with analyses via flow cytometry. We also demonstrate quantitative 3D microscopic imaging of biomarker expression in tissue models and in thick human biopsy samples of normal, HER2-negative, and HER2-positive breast tumors. This strategy enables rapid, quantitative, and unambiguous volumetric microscopy of biomarker expression in thick tissues, including whole biopsies, and will enable real-time optical assessment of disease markers in the living body. PMID- 19289066 TI - Nanoscale probing reveals that reduced stiffness of clots from fibrinogen lacking 42 N-terminal Bbeta-chain residues is due to the formation of abnormal oligomers. AB - Removal of Bbetal-42 from fibrinogen by Crotalus atrox venom results in a molecule lacking fibrinopeptide B and part of a thrombin binding site. We investigated the mechanism of polymerization of desBbeta1-42 fibrin. Fibrinogen trinodular structure was clearly observed using high resolution noncontact atomic force microscopy. E-regions were smaller in desBbeta1-42 than normal fibrinogen (1.2 nm +/- 0.3 vs. 1.5 nm +/- 0.2), whereas there were no differences between the D-regions (1.7 nm +/- 0.4 vs. 1.7 nm +/- 0.3). Polymerization rate for desBbeta1-42 was slower than normal, resulting in clots with thinner fibers. Differences in oligomers were found, with predominantly lateral associations for desBbeta1-42 and longitudinal associations for normal fibrin. Clot elasticity as measured by magnetic tweezers showed a G' of approximately 1 Pa for desBbeta1-42 compared with approximately 8 Pa for normal fibrin. Spring constants of early stage desBbeta1-42 single fibers determined by atomic force microscopy were approximately 3 times less than normal fibers of comparable dimensions and development. We conclude that Bbeta1-42 plays an important role in fibrin oligomer formation. Absence of Bbeta1-42 influences oligomer structure, affects the structure and properties of the final clot, and markedly reduces stiffness of the whole clot as well as individual fibrin fibers. PMID- 19289067 TI - Mitochondrial creatine kinase binding to phospholipid monolayers induces cardiolipin segregation. AB - It is well established that the octameric mitochondrial form of creatine kinase (mtCK) binds to the outer face of the inner mitochondrial membrane mainly via electrostatic interactions with cardiolipin (CL). However, little is known about the consequences of these interactions on membrane and protein levels. Brewster angle microscopy investigations provide, for the first time to our knowledge, images indicating that mtCK binding induced cluster formation on CL monolayers. The thickness of the clusters (10-12 nm) corresponds to the theoretical height of the mtCK-CL complex. Protein insertion into a condensed CL film, together with monolayer stabilization after protein addition, was observed by means of differential capacity measurements. Polarization modulation infrared reflection absorption spectroscopy showed that the mean orientation of alpha-helices within the protein shifted upon CL binding from 30 degrees to 45 degrees with respect to the interface plane, demonstrating protein domain movements. A comparison of data obtained with CL and phosphatidylcholine/phosphatidylethanolamine/CL (2:1:1) monolayers indicates that mtCK is able to selectively recruit CL molecules within the mixed monolayer, consolidating and changing the morphology of the interfacial film. Therefore, CL-rich domains induced by mtCK binding could modulate mitochondrial inner membrane morphology into a raft-like organization and influence essential steps of mitochondria-mediated apoptosis. PMID- 19289068 TI - Logarithmic sensing in Escherichia coli bacterial chemotaxis. AB - We studied the response of swimming Escherichia coli (E. coli) bacteria in a comprehensive set of well-controlled chemical concentration gradients using a newly developed microfluidic device and cell tracking imaging technique. In parallel, we carried out a multi-scale theoretical modeling of bacterial chemotaxis taking into account the relevant internal signaling pathway dynamics, and predicted bacterial chemotactic responses at the cellular level. By measuring the E. coli cell density profiles across the microfluidic channel at various spatial gradients of ligand concentration grad[L] and the average ligand concentration [L] near the peak chemotactic response region, we demonstrated unambiguously in both experiments and model simulation that the mean chemotactic drift velocity of E. coli cells increased monotonically with grad [L]/[L] or approximately grad(log[L])--that is E. coli cells sense the spatial gradient of the logarithmic ligand concentration. The exact range of the log-sensing regime was determined. The agreements between the experiments and the multi-scale model simulation verify the validity of the theoretical model, and revealed that the key microscopic mechanism for logarithmic sensing in bacterial chemotaxis is the adaptation kinetics, in contrast to explanations based directly on ligand occupancy. PMID- 19289070 TI - Role of E-cadherin in membrane-cortex interaction probed by nanotube extrusion. AB - This study aims to define the role of E-cadherin (Ecad) engagement in cell-cell contact during membrane-cortex interaction. As a tool, we used a hydrodynamic membrane tube extrusion technique to characterize the mechanical interaction between the plasma membrane and the underlying cortical cytoskeleton. Cells were anchored on 4.5 microm beads coated with polylysine (PL) to obtain nonspecific cell adhesion or with an antibody against Ecad to mimic specific Ecad-mediated cell adhesion. We investigated tube length dynamics L(t) over time and through successive extrusions applied to the cell at regular time intervals. A constant slow velocity was observed for the first extrusion, for PL-attached cells. Subsequent extrusions had two phases: an initial high-velocity regime followed by a low-velocity regime. Successive extrusions gradually weakened the binding of the membrane around the tube neck to the underlying cortical cytoskeleton. Cells specifically attached via Ecad first exhibited a very low extrusion velocity regime followed by a faster extrusion regime similar to nonspecific extrusion. This indicates that Ecad strengthens the membrane-cortical cytoskeleton interaction, but only in a restricted area corresponding to the site of contact between the cell and the bead. Occasional giant "cortex" tubes were extruded with specifically anchored cells, demonstrating that the cortex remained tightly bound to the membrane through Ecad-mediated adhesion at the contact site. PMID- 19289069 TI - Action potential modulates Ca2+-dependent and Ca2+-independent secretion in a sensory neuron. AB - Neurotransmitter release normally requires calcium triggering. However, the somata of dorsal root ganglion (DRG) neurons possess a calcium-independent but voltage-dependent secretion (CIVDS) in addition to the classic calcium-dependent secretion (CDS). Here, we investigated the physiological role of CIVDS and the contributions of CIVDS and CDS induced by action potentials (APs) in DRG soma. Using membrane capacitance measurements, caged calcium photolysis, and membrane capacitance kinetics analysis, we demonstrated that AP-induced secretion had both CIVDS and CDS components. Following physiological stimuli, the dominant component of AP-induced secretion was either CIVDS for spontaneous firing or CDS for high intensity stimuli. AP frequency modulates CDS-coupled exocytosis and CIVDS coupled endocytosis but not CIVDS-coupled exocytosis and CDS-coupled endocytosis. Finally, CIVDS did not contribute to excitatory postsynaptic currents induced by APs in DRG presynaptic terminals in the spinal cord. Thus, CIVDS is probably an essential physiological component of AP-induced secretion in the soma. These findings bring novel insights into primary sensory processes in DRG neurons. PMID- 19289071 TI - Control and regulation of mitochondrial energetics in an integrated model of cardiomyocyte function. AB - Understanding the regulation and control of complex networks of reactions requires analytical tools that take into account the interactions between individual network components controlling global network function. Here, we apply a generalized matrix method of control analysis to calculate flux and concentration control coefficients, as well as response coefficients, in an integrated model of excitation-contraction (EC) coupling and mitochondrial energetics (ECME model) in the cardiac ventricular myocyte. Control and regulation of oxygen consumption (V(O2)) was first assessed in a mitochondrion model, and then in the integrated cardiac myocyte model under resting and working conditions. The results demonstrate that in the ECME model, control of respiration is distributed among cytoplasmic ATPases and mitochondrial processes. The magnitude of control by cytoplasmic ATPases increases under working conditions. The model prediction that the respiratory chain exerts strong positive control on V(O2) (control coefficient 0.89) was corroborated experimentally in cardiac trabeculae utilizing the inhibitor titration method. In the model, mitochondrial respiration displayed the highest response coefficients with respect to the concentration of cytoplasmic ATP. This was due to the high elasticity of ANT flux toward ATP in the cytoplasm. The analysis reveals the complex interdependence of sarcolemmal, cytoplasmic, and mitochondrial processes that contribute to the control of energy supply and demand in the heart. Moreover, by visualizing the structure of control of the metabolic network of the myocyte, we provide support for the emerging concept of control by diffuse loops, in which action on the network (e.g., by a pharmacological agent) may bring about changes in processes without obvious direct mechanistic links between them. PMID- 19289072 TI - Concentration gradient effects of sodium and lithium ions and deuterium isotope effects on the activities of H+-ATP synthase from chloroplasts. AB - We explored the concentration gradient effects of the sodium and lithium ions and the deuterium isotope's effects on the activities of H(+)-ATP synthase from chloroplasts (CF(0)F(1)). We found that the sodium concentration gradient can drive the ATP synthesis reaction of CF(0)F(1). In contrast, the lithium ion can be an efficient enzyme-inhibitor by blocking the entrance channel of the ion translocation pathway in CF(0). In the presence of sodium or lithium ions and with the application of a membrane potential, unexpected enzyme behaviors of CF(0)F(1) were evident. To account for these observations, we propose that both of the sodium and lithium ions could undergo localized hydrolysis reactions in the chemical environment of the ion channel of CF(0). The protons generated locally could proceed to complete the ion translocation process in the ATP synthesis reaction of CF(0)F(1). Experimental and theoretical deuterium isotope effects of the localized hydrolysis on the activities of CF(0)F(1), and the energetics of these related reactions, support this proposed mechanism. Our experimental observations could be understood in the framework of the well established ion translocation models for the H(+)-ATP synthase from Escherichia coli, and the Na(+)-ATP synthase from Propionigenium modestum and Ilyobacter tartaricus. PMID- 19289073 TI - A novel kinetic assay of mitochondrial ATP-ADP exchange rate mediated by the ANT. AB - A novel method exploiting the differential affinity of ADP and ATP to Mg(2+) was developed to measure mitochondrial ADP-ATP exchange rate. The rate of ATP appearing in the medium after addition of ADP to energized mitochondria, is calculated from the measured rate of change in free extramitochondrial [Mg(2+)] reported by the membrane-impermeable 5K(+) salt of the Mg(2+)-sensitive fluorescent indicator, Magnesium Green, using standard binding equations. The assay is designed such that the adenine nucleotide translocase (ANT) is the sole mediator of changes in [Mg(2+)] in the extramitochondrial volume, as a result of ADP-ATP exchange. We also provide data on the dependence of ATP efflux rate within the 6.8-7.8 matrix pH range as a function of membrane potential. Finally, by comparing the ATP-ADP steady-state exchange rate to the amount of the ANT in rat brain synaptic, brain nonsynaptic, heart and liver mitochondria, we provide molecular turnover numbers for the known ANT isotypes. PMID- 19289074 TI - Fluctuation analysis of tetanic rundown (short-term depression) at a corticothalamic synapse. AB - Hypothetical scenarios for "tetanic rundown" ("short-term depression") of synaptic signals evoked by stimulus trains differ in evolution of quantal amplitude (Q) and covariances between signals. With corticothalamic excitatory postsynaptic currents (EPSCs) evoked by 2.5- to 20-Hz trains, we found Q (estimated using various corrections of variance/mean ratios) to be unchanged during rundown and close to the size of stimulus-evoked "miniatures". Except for covariances, results were compatible with a depletion model, according to which incomplete "refill" after probabilistic quantal release entails release-site "emptying". For five neurons with 20 train repetitions at each frequency, there was little between-neuron variation of rundown; pool-refill rate increased with stimulus frequency and evolved during rundown. Covariances did not fit the depletion model or theoretical alternatives, being excessively negative for adjacent EPSCs early in trains, absent at equilibrium, and anomalously positive for some nonadjacent EPSCs. The anomalous covariances were unaltered during pharmacological blockade of receptor desensitization and saturation. These findings suggest that pool-refill rate and release probability at each release site are continually modulated by antecedent outputs in its neighborhood, possibly via feedback mechanisms. In all data sets, sampling errors for between train variances were much less than theoretical, warranting reconsideration of the probabilistic nature of quantal transmitter release. PMID- 19289075 TI - Uniform action potential repolarization within the sarcolemma of in situ ventricular cardiomyocytes. AB - Previous studies have speculated, based on indirect evidence, that the action potential at the transverse (t)-tubules is longer than at the surface membrane in mammalian ventricular cardiomyocytes. To date, no technique has enabled recording of electrical activity selectively at the t-tubules to directly examine this hypothesis. We used confocal line-scan imaging in conjunction with the fast response voltage-sensitive dyes ANNINE-6 and ANNINE-6plus to resolve action potential-related changes in fractional dye fluorescence (DeltaF/F) at the t tubule and surface membranes of in situ mouse ventricular cardiomyocytes. Peak DeltaF/F during action potential phase 0 depolarization averaged -21% for both dyes. The shape and time course of optical action potentials measured with the water-soluble ANNINE-6plus were indistinguishable from those of action potentials recorded with intracellular microelectrodes in the absence of the dye. In contrast, optical action potentials measured with the water-insoluble ANNINE-6 were significantly prolonged compared to the electrical recordings obtained from dye-free hearts, suggesting electrophysiological effects of ANNINE-6 and/or its solvents. With either dye, the kinetics of action potential-dependent changes in DeltaF/F during repolarization were found to be similar at the t-tubular and surface membranes. This study provides what to our knowledge are the first direct measurements of t-tubule electrical activity in ventricular cardiomyocytes, which support the concept that action potential duration is uniform throughout the sarcolemma of individual cells. PMID- 19289076 TI - Harness the power: new insights into the inhibition of YAP/Yorkie. AB - YAP/Yorkie (Yki) is a transcriptional coactivator that controls organ size; dysregulation causes tumorigenesis by stimulating cell proliferation and inhibiting apoptosis. The Hippo pathway restrains YAP/Yki activity, but it remains unclear how cellular regulation at the plasma membrane influences the Hippo-YAP/Yki pathway. Two papers in this issue on Developmental Cell, by Badouel et al. and Nishioka et al., address this question. PMID- 19289077 TI - Sphingosine-1 phosphate: a new player in osteoimmunology. AB - Osteoclasts, the cells that degrade bone, differentiate from bone marrow-derived myeloid precursors. Recent work by Ishii et al. shows that sphingosine-1 phosphate in blood attracts osteoclast precursors into the bloodstream to keep them away from bone surfaces. These findings point to a novel mechanism to inhibit bone degradation and prevent bone loss. PMID- 19289078 TI - Get ready to Wnt: prepatterning in neuromuscular junction formation. AB - Clustering of acetylcholine receptors (AChR) in muscle fibers prior to innervation by motor neurons is thought to be involved in neuromuscular junction formation. Jing et al. now report in Neuron that this prepatterning of AChRs, via a novel MuSK-dependent Wnt pathway, may guide motor axons to the central region of muscle fibers for synapse formation in zebrafish. PMID- 19289079 TI - Tightening the connections between cadherins and fibronectin matrix. AB - Fibronectin extracellular matrix is assembled and remodeled throughout embryogenesis and plays key roles in early vertebrate development. In this issue of Developmental Cell, Dzamba et al. reveal, through their studies of Xenopus embryos, a novel mechanism for regulating fibronectin matrix assembly through Wnt signaling and cadherin-mediated cell-cell adhesion. PMID- 19289080 TI - Tgf-beta superfamily signaling in embryonic development and homeostasis. AB - TGF-beta superfamily signaling pathways emerged with the evolution of multicellular animals, suggesting that these pathways contribute to the increased diversity and complexity required for the development and homeostasis of these organisms. In this review we begin by exploring some key developmental and disease processes requiring TGF-beta ligands to underscore the fundamental importance of these pathways before delving into the molecular mechanism of signal transduction, focusing on recent findings. Finally, we discuss how these ligands act as morphogens, how their activity and signaling range is regulated, and how they interact with other signaling pathways to achieve their specific and varied functional roles. PMID- 19289081 TI - Nuclear export of Smad2 and Smad3 by RanBP3 facilitates termination of TGF-beta signaling. AB - Smad2 and Smad3 (Smad2/3) are key intracellular signal transducers for TGF-beta signaling, and their transcriptional activities are controlled through reversible phosphorylation and nucleocytoplasmic shuttling. However, the precise mechanism underlying nuclear export of Smad2/3 remains elusive. Here we report the essential function of RanBP3 in selective nuclear export of Smad2/3 in the TGF beta pathway. RanBP3 directly recognizes dephosphorylated Smad2/3, which results from the activity of nuclear Smad phosphatases, and mediates nuclear export of Smad2/3 in a Ran-dependent manner. As a result, increased expression of RanBP3 inhibits TGF-beta signaling in mammalian cells and Xenopus embryos. Conversely, depletion of RanBP3 expression or dominant-negative inhibition of RanBP3 enhances TGFbeta-induced antiproliferative and transcriptional responses. In conclusion, our study supports a definitive role for RanBP3 in mediating Smad2/3 nuclear export and terminating TGF-beta signaling. PMID- 19289082 TI - Neurogenin3 is sufficient for transdetermination of hepatic progenitor cells into neo-islets in vivo but not transdifferentiation of hepatocytes. AB - The transcription factor Neurogenin3 (Ngn3) is required for islet-cell type specification. Here, we show that hepatic gene transfer of Ngn3 transiently induces insulin in terminally differentiated hepatocytes but fails to transdifferentiate them, i.e., switch their lineage into islet cells. However, Ngn3 leads to long-term diabetes reversal in mice due to the emergence of periportal islet-like cell clusters. These neo-islets display glycemia-regulated insulin, beta-cell-specific transcripts, and an islet-specific transcription cascade, and they produce all four major islet hormones. They appear to arise from hepatic progenitor cells, most likely endoderm-derived oval cells. Thus, transfer of a single lineage-defining transcription factor, Ngn3, is sufficient to induce cell-lineage switching from a hepatic to an islet lineage in these progenitor cells, a process consistent with transdetermination, i.e, lineage switching in lineage-determined, but not terminally differentiated, cells. This paradigm of induced transdetermination of receptive progenitor cells in vivo may be generally applicable to therapeutic organogenesis for multiple diseases, including diabetes. PMID- 19289083 TI - The human kinetochore Ska1 complex facilitates microtubule depolymerization coupled motility. AB - Mitotic chromosome segregation requires that kinetochores attach to microtubule polymers and harness microtubule dynamics to drive chromosome movement. In budding yeast, the Dam1 complex couples kinetochores with microtubule depolymerization. However, a metazoan homolog of the Dam1 complex has not been identified. To identify proteins that play a corresponding role at the vertebrate kinetochore-microtubule interface, we isolated a three subunit human Ska1 complex, including the previously uncharacterized protein Rama1 that localizes to the outer kinetochore and spindle microtubules. Depletion of Ska1 complex subunits severely compromises proper chromosome segregation. Reconstituted Ska1 complex possesses two separable biochemical activities: direct microtubule binding through the Ska1 subunit, and microtubule-stimulated oligomerization imparted by the Rama1 subunit. The full Ska1 complex forms assemblies on microtubules that can facilitate the processive movement of microspheres along depolymerizing microtubules. In total, these results demonstrate a critical role for the Ska1 complex in interacting with dynamic microtubules at the outer kinetochore. PMID- 19289084 TI - Regulation of DUOX by the Galphaq-phospholipase Cbeta-Ca2+ pathway in Drosophila gut immunity. AB - All metazoan guts are in constant contact with diverse food-borne microorganisms. The signaling mechanisms by which the host regulates gut-microbe interactions, however, are not yet clear. Here, we show that phospholipase C-beta (PLCbeta) signaling modulates dual oxidase (DUOX) activity to produce microbicidal reactive oxygen species (ROS) essential for normal host survival. Gut-microbe contact rapidly activates PLCbeta through Galphaq, which in turn mobilizes intracellular Ca(2+) through inositol 1,4,5-trisphosphate generation for DUOX-dependent ROS production. PLCbeta mutant flies had a short life span due to the uncontrolled propagation of an essential nutritional microbe, Saccharomyces cerevisiae, in the gut. Gut-specific reintroduction of the PLCbeta restored efficient DUOX-dependent microbe-eliminating capacity and normal host survival. These results demonstrate that the Galphaq-PLCbeta-Ca(2+)-DUOX-ROS signaling pathway acts as a bona fide first line of defense that enables gut epithelia to dynamically control yeast during the Drosophila life cycle. PMID- 19289085 TI - The Hippo signaling pathway components Lats and Yap pattern Tead4 activity to distinguish mouse trophectoderm from inner cell mass. AB - Outside cells of the preimplantation mouse embryo form the trophectoderm (TE), a process requiring the transcription factor Tead4. Here, we show that transcriptionally active Tead4 can induce Cdx2 and other trophoblast genes in parallel in embryonic stem cells. In embryos, the Tead4 coactivator protein Yap localizes to nuclei of outside cells, and modulation of Tead4 or Yap activity leads to changes in Cdx2 expression. In inside cells, Yap is phosphorylated and cytoplasmic, and this involves the Hippo signaling pathway component Lats. We propose that active Tead4 promotes TE development in outside cells, whereas Tead4 activity is suppressed in inside cells by cell contact- and Lats-mediated inhibition of nuclear Yap localization. Thus, differential signaling between inside and outside cell populations leads to changes in cell fate specification during TE formation. PMID- 19289086 TI - The FERM-domain protein Expanded regulates Hippo pathway activity via direct interactions with the transcriptional activator Yorkie. AB - The Hippo kinase pathway plays a central role in growth regulation and tumor suppression from flies to man. The Hippo/Mst kinase phosphorylates and activates the NDR family kinase Warts/Lats, which phosphorylates and inhibits the transcriptional activator Yorkie/YAP. Current models place the FERM-domain protein Expanded upstream of Hippo kinase in growth control. To understand how Expanded regulates Hippo pathway activity, we used affinity chromatography and mass spectrometry to identify Expanded-binding proteins. Surprisingly we find that Yorkie is the major Expanded-binding protein in Drosophila S2 cells. Expanded binds Yorkie at endogenous levels via WW-domain-PPxY interactions, independently of Yorkie phosphorylation at S168, which is critical for 14-3-3 binding. Expanded relocalizes Yorkie from the nucleus, abrogating its nuclear activity, and it can regulate growth downstream of warts in vivo. These data lead to a new model whereby Expanded functions downstream of Warts, in concert with 14 3-3 proteins to sequester Yorkie in the cytoplasm, inhibiting growth activity of the Hippo pathway. PMID- 19289087 TI - Cadherin adhesion, tissue tension, and noncanonical Wnt signaling regulate fibronectin matrix organization. AB - In this study we demonstrate that planar cell polarity signaling regulates morphogenesis in Xenopus embryos in part through the assembly of the fibronectin (FN) matrix. We outline a regulatory pathway that includes cadherin adhesion and signaling through Rac and Pak, culminating in actin reorganization, myosin contractility, and tissue tension, which, in turn, directs the correct spatiotemporal localization of FN into a fibrillar matrix. Increased mechanical tension promotes FN fibril assembly in the blastocoel roof (BCR), while reduced BCR tension inhibits matrix assembly. These data support a model for matrix assembly in tissues where cell-cell adhesions play an analogous role to the focal adhesions of cultured cells by transferring to integrins the tension required to direct FN fibril formation at cell surfaces. PMID- 19289088 TI - Erbin and the NF2 tumor suppressor Merlin cooperatively regulate cell-type specific activation of PAK2 by TGF-beta. AB - Transforming growth factor beta (TGF-beta) family ligands are pleotropic proteins with diverse cell-type-specific effects on growth and differentiation. For example, PAK2 activation is critical for the proliferative/profibrotic action of TGF-beta on mesenchymal cells, and yet it is not responsive to TGF-beta in epithelial cells. We therefore investigated the regulatory constraints that prevent inappropriate PAK2 activation in epithelial cultures. The results show that the epithelial-enriched protein Erbin controls the function of the NF2 tumor suppressor Merlin by determining the output of Merlin's physical interactions with active PAK2. Whereas mesenchymal TGF-beta signaling induces PAK2-mediated inhibition of Merlin function in the absence of Erbin, Erbin/Merlin complexes bind and inactivate GTPase-bound PAK2 in epithelia. These results not only identify Erbin as a key determinant of epithelial resistance to TGF-beta signaling, they also show that Erbin controls Merlin tumor suppressor function by switching the functional valence of PAK2 binding. PMID- 19289089 TI - Annexin A2-dependent polymerization of actin mediates endosome biogenesis. AB - Early endosomes give rise to multivesicular intermediates during transport toward late endosomes. Much progress has been made in understanding the sorting of receptors into these intermediates, but the mechanisms responsible for their biogenesis remain unclear. Here, we report that F-actin is necessary for transport beyond early endosomes and endosome formation. We found that endosomes captured by actin cables were essentially stationary, but early endosomes also exhibited patches of F-actin and facilitated selective F-actin nucleation and polymerization. Our data show that nucleation of actin patches by early endosomes is strictly dependent on annexin A2, a protein involved in early-to-late endosome transport. It also requires the actin nucleation factor Spire1 and involves Arp2/3, which is needed for filament branching. We conclude that actin patches are nucleated on early endosomes via annexin A2 and Spire1, and that these patches control endosome biogenesis, presumably by driving the membrane remodeling process. PMID- 19289090 TI - Intrinsic tumor suppression and epithelial maintenance by endocytic activation of Eiger/TNF signaling in Drosophila. AB - Oncogenic alterations in epithelial tissues often trigger apoptosis, suggesting an evolutionary mechanism by which organisms eliminate aberrant cells from epithelia. In Drosophila imaginal epithelia, clones of cells mutant for tumor suppressors, such as scrib or dlg, lose their polarity and are eliminated by cell death. Here, we show that Eiger, the Drosophila tumor necrosis factor (TNF), behaves like a tumor suppressor that eliminates oncogenic cells from epithelia through a local endocytic JNK-activation mechanism. In the absence of Eiger, these polarity-deficient clones are no longer eliminated; instead, they grow aggressively into tumors. We show that in scrib clones endocytosis is elevated, which translocates Eiger to endocytic vesicles and leads to activation of apoptotic JNK signaling. Furthermore, blocking endocytosis prevents both JNK activation and cell elimination. Our data indicate that TNF signaling and the endocytic machinery could be components of an evolutionarily conserved fail-safe mechanism by which animals protect against neoplastic development. PMID- 19289091 TI - NA-Seq: a discovery tool for the analysis of chromatin structure and dynamics during differentiation. AB - It is well established that epigenetic modulation of genome accessibility in chromatin occurs during biological processes. Here we describe a method based on restriction enzymes and next-generation sequencing for identifying accessible DNA elements using a small amount of starting material, and use it to examine myeloid differentiation of primary human CD34+ cells. The accessibility of several classes of cis-regulatory elements was a predictive marker of in vivo DNA binding by transcription factors, and was associated with distinct patterns of histone posttranslational modifications. We also mapped large chromosomal domains with differential accessibility in progenitors and maturing cells. Accessibility became restricted during differentiation, correlating with a decreased number of expressed genes and loss of regulatory potential. Our data suggest that a permissive chromatin structure in multipotent cells is progressively and selectively closed during differentiation, and illustrate the use of our method for the identification of functional cis-regulatory elements. PMID- 19289092 TI - A radioisotope label-free alpha-bungarotoxin-binding assay using BIAcore sensor chip technology for real-time analysis. AB - alpha-Bungarotoxin (alpha-bgtx)-binding proteins, including certain nicotinic acetylcholine receptors and acetylcholine-binding proteins (AChBPs), are frequently characterized with radioisotope-labeled alpha-bgtx-binding assays. Such assays, however, preclude investigations of binding interactions in real time and are hampered by the inconveniences associated with radioisotope-labeled reagents. We used surface plasmon resonance-based technology (BIAcore) to investigate the binding of recombinant AChBP to CM-5 sensor chip surfaces with directly immobilized alpha-bgtx. We validated our BIAcore results by comparing the same biological samples using the traditional (125)I-labeled alpha-bgtx binding assay. An alpha-bgtx sensor chip, as described here, enables detailed, real-time, radioisotope-free interaction studies that can greatly facilitate the characterization of novel alpha-bgtx-binding proteins and complexes. PMID- 19289093 TI - Identification of a simple and sensitive microplate method for the detection of oversulfated chondroitin sulfate in heparin products. AB - Heparin is a commonly implemented anticoagulant used to treat critically ill patients. Recently, a number of commercial lots of heparin products were found to be contaminated with an oversulfated chondroitin sulfate (OSCS) derivative that could elicit a hypotensive response in pigs following a single high-dose infusion. Using both contaminated heparin products and the synthetically produced derivative, we showed that the OSCS produces dose-dependent hypotension in pigs. The no observed effect level (NOEL) for this contaminant appears to be approximately 1mg/kg, corresponding to a contamination level of approximately 3%. We also demonstrated that OSCS can be identified in heparin products using a simple, inexpensive, commercially available heparin enzyme immunoassay (EIA) kit that has a limit of detection of approximately 0.1%, well below the NOEL. This kit may provide a useful method to test heparin products for contamination with oversulfated GAG derivatives. PMID- 19289094 TI - Linear and accurate quantitation of proenkephalin-derived peptides by isotopic labeling with internal standards and mass spectrometry. AB - Proenkephalin (PE) represents the precursor protein of the active peptide neurotransmitter enkephalin. Quantitative analysis of peptides and proteins is an objective of mass spectrometry-based studies of biological systems and will be important for studying the proteolytic conversion of proproteins to active enkephalin and neuropeptides. The goal of this study was to define and optimize quantitation of different amounts of tryptic peptides derived from PE using light (H4, 4 hydrogens) and heavy (D4, 4 deuteriums) succinic anhydride for isotopic labeling of peptides analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Comparisons were made between PE-derived peptides with and without internal standards. Importantly, incorporation of internal standards of known amounts of heavy isotope-labeled tryptic peptides of PE provided linear calibration plots with accurate quantitation. In contrast, comparison of light and heavy isotope-labeled peptides without internal standards produced variable and inaccurate nonlinear isotopic ratio comparisons of PE-derived peptides. These results demonstrate that use of internal standards composed of a defined amount(s) of standard peptides (PE-derived tryptic peptides) is necessary for high-quality linear quantitation of peptides by isotopic labeling and MS/MS. PMID- 19289095 TI - A therapeutic antibody and its antigen form different complexes in serum than in phosphate-buffered saline: a study by analytical ultracentrifugation. AB - During the development of protein therapeutics, characterization of the active pharmaceutical ingredient is performed extensively to ensure the stability, safety, and efficacy of the drug. Little is known, however, about the characteristics of protein drugs circulating in the blood. The recent availability of a fluorescence detection system (FDS) in analytical ultracentrifugation (AUC) instruments enables the characterization of fluorescently labeled proteins in biological fluids. AUC provides information about protein size, shape, self-association, and binding while avoiding many limitations associated with size exclusion chromatography. Furthermore, with the specificity and sensitivity of FDS, measurements can be performed at physiological concentrations directly in serum. In the current study, we used omalizumab, an anti-immunoglobulin E (IgE) monoclonal antibody, to demonstrate the potential of using AUC-FDS for the study of a monoclonal antibody and its complexes directly in human serum. Omalizumab properties were essentially unaltered after labeling with the fluorescent dye Alexa Fluor 488. In addition, omalizumab and IgE formed different complexes in serum than in phosphate-buffered saline in terms of both size and affinity. PMID- 19289096 TI - Functional complementation of the Arabidopsis thaliana psbo1 mutant phenotype with an N-terminally His6-tagged PsbO-1 protein in photosystem II. AB - The Arabidopsis thaliana mutant psbo1 has recently been described and characterized. Loss of expression of the PsbO-1 protein leads to a variety of functional perturbations including elevated levels of the PsbO-2 protein and defects on both the oxidizing- and reducing-sides of Photosystem II. In this communication, two plant lines were produced using the psbo1 mutant as transgenic host, which contained an N-terminally histidine(6)-tagged PsbO-1 protein. This protein was expressed and correctly targeted into the thylakoid lumen. Immunological analysis indicated that different levels of expression of the modified PsbO-1 protein were obtained in different transgenic plant lines and that the level of expression in each line was stable over several generations. Examination of the Photosystem II closure kinetics demonstrated that the defective double reduction of Q(B) and the delayed exchange of Q(B)H(2) with the plastoquinone pool which were observed during the characterization of the psbo1 mutant were effectively restored to wild-type levels by the His(6)-tagged PsbO-1 protein. Flash fluorescence induction and decay were also examined. Our results indicated that high expression of the modified PsbO-1 was required to increase the ratio of PS II(alpha)/PS II(beta) reaction centers to wild-type levels. Fluorescence decay kinetics in the absence of DCMU indicated that the expression of the His(6)-tagged PsbO-1 protein restored efficient electron transfer to Q(B), while in the presence of DCMU, charge recombination between Q(A)(-) and the S(2) state of the oxygen-evolving complex occurred at near wild-type rates. Our results indicate that high expression of the His(6)-tagged PsbO-1 protein efficiently complements nearly all of the photochemical defects observed in the psbo1 mutant. Additionally, this study establishes a platform on which the in vivo consequences of site-directed mutagenesis of the PsbO-1 protein can be examined. PMID- 19289097 TI - Identification of a novel human endogenous retrovirus and promoter activity of its 5' U3. AB - Up-regulation of human endogenous retroviruses (HERVs) sometimes occurs in cancer. This study identifies a novel HERV (HERV-HX) in colon tumor tissues, which is an H family HERV (HERV-H) element located on chromosome Xp22.32. The full-length transcript sequence was identified and characterized. Quantitative RT PCR indicated it is up-regulated in colon tumor samples. Using RT-PCR to analyze the transcription of the env-deleted HERV-HX and other env-intact HERV-H elements, it was demonstrated that transcription of HERV-H in colon cancer is not associated with the env gene. A 17-bp sequence was found within the 5' U3 region of HERV-HX, and only 8-bp sequences existed on its homologous U3 regions in the corresponding loci. Promoter activity assays indicated that replacing the 17-bp sequence with an 8-bp sequence or deleting it reduced its activity, suggesting that the 17-bp sequence is important to the expression of HERV-HX. PMID- 19289098 TI - Sifuvirtide, a potent HIV fusion inhibitor peptide. AB - Enfuvirtide (ENF) is currently the only FDA approved HIV fusion inhibitor in clinical use. Searching for more drugs in this category with higher efficacy and lower toxicity seems to be a logical next step. In line with this objective, a synthetic peptide with 36 amino acid residues, called Sifuvirtide (SFT), was designed based on the crystal structure of gp41. In this study, we show that SFT is a potent anti-HIV agent with relatively low cytotoxicity. SFT was found to inhibit replication of all tested HIV strains. The effective concentrations that inhibited 50% viral replication (EC(50)), as determined in all tested strains, were either comparable or lower than benchmark values derived from well-known anti-HIV drugs like ENF or AZT, while the cytotoxic concentrations causing 50% cell death (CC(50)) were relatively high, rendering it an ideal anti-HIV agent. A GST-pull down assay was performed to confirm that SFT is a fusion inhibitor. Furthermore, the activity of SFT on other targets in the HIV life cycle was also investigated, and all assays showed negative results. To further understand the mechanism of action of HIV peptide inhibitors, resistant variants of HIV-1(IIIB) were derived by serial virus passage in the presence of increasing doses of SFT or ENF. The results showed that there was cross-resistance between SFT and ENF. In conclusion, SFT is an ideal anti-HIV agent with high potency and low cytotoxicity, but may exhibit a certain extent of cross-resistance with ENF. PMID- 19289099 TI - A conserved hydrogen-bond network stabilizes the structure of Beta class glutathione S-transferases. AB - We identified a network of hydrogen bonds that is conserved in the structures of bacterial Beta class glutathione S-transferases (GSTs). It is formed by three residues: a serine, a histidine and a glutamate, together with a water molecule that links the serine with the histidine. This network connects the first helix of the N-terminal glutaredoxin-like domain with the last helix of the C-terminal GST-specific all helical domain. Here we show that substitution of Ochrobactrum anthropi GST His15 and Glu198 with alanine greatly compromises the catalytic efficiency of the enzyme, even though none of these residues takes part to the enzyme active site. Thermal and chemical denaturation experiments point to a role for this network in global structure stabilization. Furthermore, we show that OaGST structure looses compactness at alkanine pHs and that this behavior may be ascribed to partial disruption of the H-bond network, pointing to an important role in zippering the N-terminal and C-terminal domains of the protein. PMID- 19289100 TI - Differential expression of stromal cell-derived factor 1 in human brain microvascular endothelial cells and pericytes involves histone modifications. AB - Stromal cell-derived factor 1 (SDF-1) regulates neovascularization, which is coordinately controlled by endothelial cells (EC) and their surrounding cells, pericytes or smooth muscle cells. In the basal state, SDF-1 expression is much lower in EC than in their surrounding cells. In this study, we evaluated epigenetic regulation to determine if it is involved in the mechanism responsible for the differential expression of SDF-1 in two types of vascular cells, brain microvascular EC (HBMEC) and pericytes (HBMP). We found that HBMEC did not express SDF-1, but that HBMP did. Furthermore, treatment of EC with 5-aza-2' dexoycytidine and trichostatin A resulted in a remarkable restoration of SDF-1 expression. Additionally, bisulfite-sequencing analysis revealed no differences in the methylation state of SDF-1 promoter between HBMEC and HBMP. Finally, a chromatin immunoprecipitation assay revealed reduced levels of histone H3 lysine 9 (H3K9) acetylation and H3K4 trimethylation with concomitant enhancement of H3K9 trimethylation in HBMEC relative to HBMP, which suggests that histone modifications are involved in the cell-specific expression of SDF-1. PMID- 19289101 TI - Inhibition of HSP27 phosphorylation by a cell-permeant MAPKAP Kinase 2 inhibitor. AB - Heat shock protein 27 (HSP27) has been implicated in many intracellular signaling processes. Since the phosphorylation of HSP27 can modulate its activity, the ability to inhibit phosphorylation of HSP27 might have clinical relevance especially with regard to the treatment of fibrosis. We have developed a cell permeant peptide inhibitor of MAPKAP Kinase 2 (MK2), an enzyme that phosphorylates HSP27, by combining a previously described peptide substrate of MK2 with a cell penetrating peptide. This novel MK2 inhibitor (MK2i) reduced HSP27 phosphorylation by MK2 in vitro. At 10 microM, MK2i inhibited TGF-beta1 induced HSP27 phosphorylation in serum-starved human keloid fibroblasts. In addition, 10 microM MK2i decreased TGF-beta1-induced expression of connective tissue growth factor and collagen type I within serum-starved keloid fibroblasts. Thus, MK2i represents a potential therapeutic for the treatment of fibrotic disorders. PMID- 19289102 TI - Possible involvement of mitogen-activated protein kinase phosphatase-1 (MKP-1) in thyrotropin-releasing hormone (TRH)-induced prolactin gene expression. AB - The role of extracellular signal-regulated kinase (ERK) in mediating the ability of thyrotropin-releasing hormone (TRH) to stimulate the prolactin gene has been well elucidated. ERK is inactivated by a dual specificity phosphatase, mitogen activated protein kinase phosphatase (MKP). In this study, we examined the induction of MKP-1 protein by thyrotropin-releasing hormone (TRH) in pituitary GH3 cells, and investigated the possible role for MKP-1 in TRH-induced prolactin gene expression. MKP-1 protein was induced significantly from 60 min after TRH stimulation, and remained elevated at 4h. The effect of TRH on MKP-1 expression was completely prevented in the presence of the MEK inhibitor, U0126. In the experiments using triptolide, a potent blocker for MKP-1, MKP-1 induction by TRH was completely inhibited in a dose-dependent manner. TRH-induced ERK activation was significantly enhanced in this condition. Prolactin promoter activity, activated by TRH, was reduced to the control level in the presence of triptolide in a dose-dependent manner. In GH3 cells, which were transfected with MKP-1 specific siRNA, both the basal and TRH-stimulated activities of the prolactin promoter were significantly reduced compared to the cells transfected with negative control siRNA. Our present results support a critical role of MKP-1 in TRH-induced, ERK-dependent, prolactin gene expression. PMID- 19289103 TI - Analysis of xylosyltransferase II binding to the anticoagulant heparin. AB - The key enzymes in the biosynthetic pathway of glycosaminoglycan production are represented by the human xylosyltransferase I and its isoform II (XylT-I and XylT II). The glycosaminoglycan heparin interacts with a variety of proteins, thereby regulating their activities, also those of xylosyltransferases. The identification of unknown amino acids responsible for heparin-binding of XylT-II was addressed in this study. Thus, six XylT-II fragments were designed as fusion proteins with MBP and we received soluble and purified MBP/XylT-II from Escherichia coli. Heparin-binding studies showed that all fragments bound with low affinity to heparin. Prolonging of XylT-II fragments did not account for a cooperative effect of multiple heparin-binding motifs and in turn for a stronger heparin-binding. Sequence alignment and surface polarity plot led to the identification of two highly positively charged Cardin-Weintraub motifs with surface accessibility, resulting in combination with short clusters of basic amino acids for strong heparin-binding of native xylosyltransferases. PMID- 19289104 TI - Skin atrophy in cytoplasmic SOD-deficient mice and its complete recovery using a vitamin C derivative. AB - Intrinsic skin ageing is characterized by atrophy and loss of elasticity. Although the skin hypertrophy induced by photoageing has been studied, the molecular mechanisms of skin atrophy during ageing remain unclear. Here, we report that copper/zinc superoxide dismutase (CuZn-SOD)-deficient mice show atrophic morphology in their skin. This atrophy is accompanied by the degeneration of collagen and elastic fibers, and skin hydroxyproline is also significantly reduced in deficient mice. These imply that the dysfunction of collagen and elastin biosynthesis are involved in the progression of skin thinning. Furthermore, transdermal administration of a vitamin C derivative which can permeate through the membrane, completely reversed the skin thinning and deterioration of collagen and elastin in the mutant mice. These indicate that the vitamin C derivative is a powerful agent for alleviating skin ageing through regeneration of collagen and elastin. The CuZn-SOD-deficient mice might be applicable to evaluation of therapeutic medicines against skin ageing. PMID- 19289105 TI - YM201636, an inhibitor of retroviral budding and PIKfyve-catalyzed PtdIns(3,5)P2 synthesis, halts glucose entry by insulin in adipocytes. AB - Silencing of PIKfyve, the sole enzyme for PtdIns(3,5)P(2) biosynthesis that controls proper endosome dynamics, inhibits retroviral replication. A novel PIKfyve-specific inhibitor YM201636 disrupts retroviral budding at 800 nM, suggesting its potential use as an antiretroviral therapeutic. Because PIKfyve is also required for optimal insulin activation of GLUT4 surface translocation and glucose influx, we tested the outcome of YM201636 application on insulin responsiveness in 3T3L1 adipocytes. YM201636 almost completely inhibited basal and insulin-activated 2-deoxyglucose uptake at doses as low as 160 nM, with IC(50)=54+/-4 nM for the net insulin response. Insulin-induced GLUT4 translocation was partially inhibited at substantially higher doses, comparable to those required for inhibition of insulin-induced phosphorylation of Akt/PKB. In addition to PIKfyve, YM201636 also completely inhibited insulin-dependent activation of class IA PI 3-kinase. We suggest that apart from PIKfyve, there are at least two additional targets for YM201636 in the context of insulin signaling to GLUT4 and glucose uptake: the insulin-activated class IA PI 3-kinase and a here-unidentified high-affinity target responsible for the greater inhibition of glucose entry vs. GLUT4 translocation. The profound inhibition of the net insulin effect on glucose influx at YM201636 doses markedly lower than those required for efficient retroviral budding disruption warns of severe perturbations in glucose homeostasis associated with potential YM201636 use in antiretroviral therapy. PMID- 19289106 TI - Inhibition of macroautophagy by bafilomycin A1 lowers proliferation and induces apoptosis in colon cancer cells. AB - Macroautophagy is a process by which cytoplasmic content and organelles are sequestered by double-membrane bound vesicles and subsequently delivered to lysosomes for degradation. Macroautophagy serves as a major intracellular pathway for protein degradation and as a pro-survival mechanism in time of stress by generating nutrients. In the present study, bafilomycin A(1), a vacuolar type H(+)-ATPase inhibitor, suppresses macroautophagy by preventing acidification of lysosomes in colon cancer cells. Diminished macroautophagy was evidenced by the accumulation of undegraded LC3 protein. Suppression of macroautophagy by bafilomycin A(1) induced G(0)/G(1) cell cycle arrest and apoptosis which were accompanied by the down-regulation of cyclin D(1) and cyclin E, the up-regulation of p21(Cip1) as well as cleavages of caspases-3, -7, -8, and -9 and PARP. Further investigation revealed that bafilomycin A(1) increased the phosphorylation of ERK, JNK, and p38. In this regard, p38 inhibitor partially reversed the anti proliferative effect of bafilomycin A(1). To conclude, inhibition of macroautophagy by bafilomycin A(1) lowers G(1)-S transition and induces apoptosis in colon cancer cells. Our results not only indicate that inhibitors of macroautophagy may be used therapeutically to inhibit cancer growth, but also delineate the relationship between macroautophagy and apoptosis. PMID- 19289107 TI - Southeast Asian AE1 associated renal tubular acidosis: cation leak is a class effect. AB - Anion Exchanger 1 (AE1) is present in the erythrocyte and also in the alpha intercalated cell; different mutations can cause either red cell disease or distal renal tubular acidosis (dRTA). Recently, we described a cation leak property in four dRTA-causing AE1 mutants, three autosomal dominant (AD) European mutants, one autosomal recessive (AR) from Southeast Asia, G701D. G701D had a very large leak property and is unusually common in SE Asia. We hypothesized that this property might confer a survival advantage. We characterized three other AR dRTA-associated AE1 mutants found in SE Asia, S773P, Delta850 and A858D via transport experiments in AE1-expressing Xenopus oocytes. These three SE Asian mutants also had cation leaks of similar magnitude to that seen in G701D, a property that distinguishes them as a discrete group. The clustering of these cation-leaky AE1 mutations to malarious areas of SE Asia suggests that they may confer malaria resistance. PMID- 19289108 TI - Carnosic acid and carnosol inhibit adipocyte differentiation in mouse 3T3-L1 cells through induction of phase2 enzymes and activation of glutathione metabolism. AB - In the previous studies, we reported that carnosic acid (CA) and carnosol (CS) originating from rosemary protected cortical neurons by activating the Keap1/Nrf2 pathway, which activation was initiated by S-alkylation of the critical cysteine thiol of the Keap1 protein by the "electrophilic"quinone-type of CA or CS. Here, we found that CA and CS inhibited the in vitro differentiation of mouse preadipocytes, 3T3-L1 cells, into adipocytes. In contrast, other physiologically active and rosemary-originated compounds were completely negative. These actions seemed to be mediated by activation of the antioxidant-response element (ARE) and induction of phase2 enzymes. This estimation is justified by our present findings that only CA and CS among rosemary-originated compounds significantly activated the ARE and induced the phase2 enzymes. Next, we performed cDNA microarray analysis in order to identify the gene(s) responsible for these biological actions and found that phase2 enzymes (Gsta2, Gclc, Abcc4, and Abcc1), all of which are involved in the metabolism of glutathione (GSH), constituted 4 of the top 5 CA-induced genes. Furthermore, CA and CS, but not the other compounds tested, significantly increased the intracellular level of total GSH. Thus, we propose that the stimulation of GSH metabolism may be a critical step for the inhibition of adipocyte differentiation in 3T3-L1 cells and suggest that pro electrophilic compounds such as CA and CS may be potential drugs against obesity related diseases. PMID- 19289109 TI - Analysis of in vitro SUMOylation using bioluminescence resonance energy transfer (BRET). AB - We demonstrated in vitro small ubiquitin-like modifier (SUMO)-mediated modification (SUMOylation) of RanGTPase activating protein-1 (RanGAP1) by using bioluminescence resonance energy transfer (BRET) for studying protein interactions. Renilla luciferase (Rluc) was fused to SUMO, and RanGAP1, the binding partner of SUMO, was fused to enhanced yellow fluorescence protein (EYFP). Upon binding of SUMO and RanGAP1, BRET was observed between EYFP (donor) and Rluc (acceptor) in the presence of E1 (Aos1/Uba2) and E2 (Ubc9) enzymes, whereas mutation (K524A) of RanGAP1 at its SUMO binding site prevented significant energy transfer. Comparing BRET and fluorescence resonance energy transfer (FRET) efficiencies using this in vitro model system, we observed that BRET efficiency was 3-fold higher than FRET efficiency, due to the lower background signal intensity of EYFP in the BRET system. Consequently, BRET system is expected to be useful for in vitro analysis of SUMOylation as well as studying other protein interactions. PMID- 19289110 TI - Src supports UDP-glucuronosyltransferase-2B7 detoxification of catechol estrogens associated with breast cancer. AB - Mammary gland-distributed and ER-bound UDP-glucuronosyltransferase (UGT)-2B7 metabolizes genotoxic catechol-estrogens (CE) associated with breast cancer initiation. Although UGT2B7 has 3 PKC- and 2 tyrosine kinase (TK)-sites, its inhibition by genistein, herbimycin-A and PP2 with parallel losses in phospho tyrosine and phospho-Y438-2B7 content indicated it requires tyrosine phosphorylation, unlike required PKC phosphorylation of UGT1A isozymes. 2B7 mutants at PKC-sites had essentially normal activity, while its TK-sites mutants, Y236F- and Y438F-2B7, were essentially inactive. Overexpression of regular or active Src, but not dominant-negative Src, in 2B7-transfected COS-1 cells increased 2B7 activity and phospho-Y438-2B7 by 50%. Co-localization of 2B7 and regular SrcTK in COS-1 cells that was dissociated by pretreatment with Src specific PP2-inhibitor provided strong evidence Src supports 2B7 activity. Consistent with these findings, evidence indicates an appropriate set of ER proteins with Src-homology binding-domains, including 2B7 and well-known multi functional Src-engaged AKAP12 scaffold, supports Src-dependent phosphorylation of CE-metabolizing 2B7 enabling it to function as a tumor suppressor. PMID- 19289111 TI - Transduction of MyoD protein into myoblasts induces myogenic differentiation without addition of protein transduction domain. AB - Protein transduction is a new technology with the potential for controlling cellular functions. Addition of tissue-specific transcription factors into cells could induce a specific differentiation pathway. In this paper, MyoD protein, a muscle-specific transcription factor, was introduced into the mouse myoblast cell line C2C12 to induce myogenic differentiation. Addition of fluorescently labeled MyoD protein revealed that MyoD can permeate the cell membrane without the addition of a protein transduction domain (PTD). Addition of MyoD protein induced formation of multi-nucleated myotubes. These results showed that MyoD protein has a PTD in its primary sequence and that myogenic differentiation can be induced by addition of MyoD protein. PMID- 19289112 TI - Estrogen receptor subtype beta2 is involved in neuromast development in zebrafish (Danio rerio) larvae. AB - Estrogens are known to play a role in both reproductive and non-reproductive functions in mammals. Estrogens and their receptors are involved in the development of the central nervous system (brain development, neuronal survival and differentiation) as well as in the development of the peripheral nervous system (sensory-motor behaviors). In order to decipher possible functions of estrogens in early development of the zebrafish sensory system, we investigated the role of estrogen receptor beta(2) (ERbeta(2)) by using a morpholino (MO) approach blocking erbeta(2) RNA translation. We further investigated the development of lateral line organs by cell-specific labeling, which revealed a disrupted development of neuromasts in morphants. The supporting cells developed and migrated normally. Sensory hair cells, however, were absent in morphants' neuromasts. Microarray analysis and subsequent in situ hybridizations indicated an aberrant activation of the Notch signaling pathway in ERbeta(2) morphants. We conclude that signaling via ERbeta(2) is essential for hair cell development and may involve an interaction with the Notch signaling pathway during cell fate decision in the neuromast maturation process. PMID- 19289113 TI - Chronic morphine administration induces over-expression of aldolase C with reduction of CREB phosphorylation in the mouse hippocampus. AB - In recent studies, alterations in the activity and expression of metabolic enzymes, such as those involved in glycolysis, have been detected in morphine dependent patients and animals. Increasing evidence demonstrates that the hippocampus is an important brain region associated with morphine dependence, but the molecular events occurring in the hippocampus following chronic exposure to morphine are poorly understood. Aldolase C is the brain-specific isoform of fructose-1, 6-bisphosphate aldolase which is a glycolytic enzyme catalyzing reactions in the glycolytic, gluconeogenic, and fructose metabolic pathways. Using Western blot and immunofluorescence assays, we found the expression of aldolase C was markedly increased in the mouse hippocampus following chronic morphine treatment. Naloxone pretreatment before morphine administration suppressed withdrawal jumping, weight loss, and overexpression of aldolase C. CREB is a transcription factor regulated through phosphorylation on Ser133, which is known to play a key role in the mechanism of morphine dependence. When detecting the expression of phosphorylated CREB (p-CREB) in the mouse hippocampus using Western blot and immunohistochemistry, we found CREB phosphorylation was clearly decreased following chronic morphine treatment. Interestingly, laser confocal microscopy showed that overexpression of aldolase C in mouse hippocampal neurons was concomitant with the decreased immunoreactivity of p-CREB. The results suggest potential links between the morphine-induced alteration of aldolase C and the regulation of CREB phosphorylation, a possible mechanism of morphine dependence. PMID- 19289114 TI - Effects of umbelliferone in a murine model of allergic airway inflammation. AB - The therapeutic effects of umbelliferone (30, 60 and 90 mg/kg), a coumarin isolated from Typha domingensis (Typhaceae) were investigated in a mouse model of bronchial asthma. BALB/c mice were immunized and challenged by nasal administration of ovalbumin. Treatment with umbelliferone (60 and 90 mg/kg) caused a marked reduction of cellularity and eosinophil numbers in bronchoalveolar lavage fluids from asthmatic mice. In addition, a decrease in mucus production and lung inflammation were observed in mice treated with umbelliferone. A reduction of IL-4, IL-5, and IL-13, but not of IFN-gamma, was found in bronchoalveolar lavage fluids of mice treated with umbelliferone, similar to that observed with dexamethasone. The levels of ovalbumin-specific IgE were not significantly altered after treatment with umbelliferone. In conclusion, our results demonstrate that umbelliferone attenuates the alteration characteristics of allergic airway inflammation. The investigation of the mechanisms of action of this molecule may contribute for the development of new drugs for the treatment of asthma. PMID- 19289115 TI - Expression and functional evidence of the prostaglandin F2alpha receptor mediating contraction in human umbilical vein. AB - Our purposes were to perform the pharmacological characterization of PGF(2alpha) receptor (prostanoid FP-receptor) involved in human umbilical vein contraction and confirm its expression in this tissue. Umbilical cords from healthy patients after full-term deliveries were employed. The vein was dissected out of cords and used for either isolated organ bath or reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assays. The natural prostanoid FP-receptor agonist, PGF(2alpha), and its selective analogues, latanoprost and bimatoprost free acids are full agonists (produce more than 80% of the maximal contractile response to 5-HT) in human umbilical vein. The agonist potency (pEC(50)) order was PGF(2alpha) (6.01+/-0.05)>latanoprost free acid (5.65+/-0.07)=bimatoprost free acid (5.59+/-0.08). The contractile effects of PGF(2alpha) and latanoprost free acid were blocked competitively by the prostanoid FP-receptor antagonist, AL 8810. The antagonist potencies (pK(B)) of AL-8810 vs. PGF(2alpha) (5.93+/-0.05) and vs. latanoprost free acid (6.40+/-0.08) in human umbilical vein are in good agreement with its ability to antagonize prostanoid FP receptors of rat, mouse and human cells. In all samples, clear signal was detected for cDNA amplification of prostanoid FP receptor and the specific prostanoid FP-receptor antibody recognized a protein of approximately 64 kDa. In conclusion, taking into account the obtained functional and biochemical data, we propose for the first time that human umbilical vein express prostanoid FP-receptors and these receptors could be involved in the vasoconstriction action of PGF(2alpha) in this tissue. PMID- 19289116 TI - Enhancement of the hypotensive effects of intrathecally injected endocannabinoids by the entourage compound palmitoylethanolamide. AB - The intrathecal (i.t.) injection of 50 and 100 nmol anandamide to urethane anesthetized rats induced a dose-dependent decrease in the mean blood pressure ( 10.6+/-1.6 mmHg and -15.0+/-1.7 mmHg, respectively; n=6) whereas a lower dose of this endocannabinoid (25 nmol) was devoid of effect. Similar responses were obtained both with the non-metabolizable analog methanandamide and with the endocannabinoid N-arachidonoyldopamine. When the sub-effective dose (25 nmol) of each compound was co-injected with palmitoylethanolamide (100 nmol), significant decreases in the blood pressure were observed (-12.3+1.3 mmHg for anandamide; 12.1+/-0.8 mmHg for methanandamide; -12.1+/-0.8 mmHg for N-arachidonoyldopamine; n=4-6). Palmitoylethanolamide also enhanced the hypotensive responses to the 50 nmol-dose of both anandamide and methanandamide. The hypotensive response induced by co-administration of palmitoylethanolamide and 25 nmol anandamide was prevented both by the cannabinoid CB(1) receptor antagonist SR 144716A (20 nmol; i.t.) and by the vanilloid TRPV1 receptor antagonist capsazepine (20 nmol; i.t.) and enhanced by pretreatment with URB602 (3.5 nmol; i.t.), a putative inhibitor of palmitoylethanolamide degradation. These results suggest that in the spinal cord palmitoylethanolamide acts as an entourage compound for the hypotensive effects of i.t. administered endocannabinoids. The facilitative action of palmitoylethanolamide affects the vanilloid TRPV1 as well as the cannabinoid CB(1) receptor-mediated effects of endocannabinoids on the blood pressure control. PMID- 19289117 TI - Elevated extracellular K+ inhibits apoptosis of corneal epithelial cells exposed to UV-B radiation. AB - The goal of this study was to determine if the high [K(+)] in tears, 20-25 mM, serves to protect corneal epithelial cells from going into apoptosis after exposure to ambient UV-B radiation. Human corneal-limbal epithelial (HCLE) cells in culture were exposed to UV-B at doses of 50-200 mJ/cm(2) followed by measurement of K(+) channel activation and activity of apoptotic pathways. Patch clamp recording showed activation of K(+) channels after UV-B exposure at 80 mJ/cm(2) or 150 mJ/cm(2) and a decrease in UV-induced K(+) efflux with increasing [K(+)](o). The UV-activated current was partially blocked by the specific K(+) channel blocker, BDS-1. DNA fragmentation, as measured by the TUNEL assay, was induced after exposure to UV-B at 100-200 mJ/cm(2). DNA fragmentation was significantly decreased when cells were incubated in 25, 50 or 100mM K(o)(+) after exposure to UV-B. The effector caspase, caspase-3, was activated by exposure to UV-B at 50-200 mJ/cm(2), but there was a significant decrease in activation when the cells were incubated in 25, 50 or 100mM K(o)(+) following exposure to UV-B. A decrease in mitochondrial potential, a possible activator of caspase-3, occurred after exposure to UV-B at 100-200 mJ/cm(2). This decrease in mitochondrial potential was prevented by 100mM K(o)(+); however, 25 or 50mM K(o)(+) provided minimal protection. Caspase-9, which is in the pathway from mitochondrial potential change to caspase-3 activation, showed little activation by UV-B radiation. Caspase-8, an initiator caspase that activates caspase-3, was activated by exposure to UV-B at 50-200 mJ/cm(2), and this UV-activation was significantly reduced by 25-100mM K(o)(+). The data show that the physiologically relevant [K(+)](o) of 25 mM can inhibit UV-B induced activation of apoptotic pathways. This suggests that the relatively high [K(+)] in tears reduces loss of K(+) from corneal epithelial cells in response to UV exposure, thereby contributing to the protection of the ocular surface from ambient UV radiation. PMID- 19289118 TI - A majority of Huntington's disease patients may be treatable by individualized allele-specific RNA interference. AB - Use of RNA interference to reduce huntingtin protein (htt) expression in affected brain regions may provide an effective treatment for Huntington disease (HD), but it remains uncertain whether suppression of both wild-type and mutant alleles in a heterozygous patient will provide more benefit than harm. Previous research has shown suppression of just the mutant allele is achievable using siRNA targeted to regions of HD mRNA containing single nucleotide polymorphisms (SNPs). To determine whether more than a minority of patients may be eligible for an allele specific therapy, we genotyped DNA from 327 unrelated European Caucasian HD patients at 26 SNP sites in the HD gene. Over 86% of the patients were found to be heterozygous for at least one SNP among those tested. Because the sites are genetically linked, one cannot use the heterozygosity rates of the individual SNPs to predict how many sites (and corresponding allele-specific siRNA) would be needed to provide at least one treatment possibility for this percentage of patients. By computing all combinations, we found that a repertoire of allele specific siRNA corresponding to seven sites can provide at least one allele specific siRNA treatment option for 85.6% of our sample. Moreover, we provide evidence that allele-specific siRNA targeting these sites are readily identifiable using a high throughput screening method, and that allele-specific siRNA identified using this method indeed show selective suppression of endogenous mutant htt protein in fibroblast cells from HD patients. Therefore, allele-specific siRNA are not so rare as to be impractical to find and use therapeutically. PMID- 19289119 TI - Anti-aggregation and fibril-destabilizing effects of sex hormones on alpha synuclein fibrils in vitro. AB - The alpha-synuclein aggregation in the brain is the hallmark of Lewy body diseases, including Parkinson's disease and dementia with Lewy bodies, and multiple system atrophy. Some epidemiological studies have revealed that estrogen therapy reduces the risk of Parkinson's disease in females. We examined the effects of estriol, estradiol, estrone, androstenedione, and testosterone on the formation and destabilization of alpha-synuclein fibrils at pH 7.5 and 37 degrees C in vitro, using fluorescence spectroscopy with thioflavin S and electron microscopy. These sex hormones, especially estriol, significantly exert anti aggregation and fibril-destabilizing effects; and hence, could be valuable preventive and therapeutic agents for alpha-synucleinopathies. PMID- 19289120 TI - Anti-melanin antibodies are increased in sera in Parkinson's disease. AB - An increasing body of research suggests that a number of immune mechanisms play a role in degenerative pathways in Parkinson's disease (PD). In the current work we investigated a posited humoral immune response in this disorder. Sera from PD patients exhibited a significantly enhanced absorbance response on a novel ELISA for anti-melanin antibodies, compared to sera from age-matched control subjects. The enhanced ELISA absorbance response was specific for catecholamine-based melanins and was unrelated to antiparkinsonian dopaminergic medication. Further, the absorbance response was significantly and negatively correlated with disease duration. These data suggest that a specific humoral anti-melanin antibody response is present in PD and is more active in early disease. While the contribution of this novel immune response to the initiation and progression of this disorder is unclear, this finding supports the hypothesis that specific immune responses occurring in PD may respond to therapeutic interventions in this disorder. PMID- 19289121 TI - Association of the eukaryotic V1VO ATPase subunits a with d and d with A. AB - Owing to the complex nature of V(1)V(O) ATPases, identification of neighboring subunits is essential for mechanistic understanding of this enzyme. Here, we describe the links between the V(1) headpiece and the V(O)-domain of the yeast V(1)V(O) ATPase via subunit A and d as well as the V(O) subunits a and d using surface plasmon resonance and fluorescence correlation spectroscopy. Binding constants of about 60 and 200 nM have been determined for the a-d and d-A assembly, respectively. The data are discussed in light of subunit a and d forming a peripheral stalk, connecting the catalytic A(3)B(3) hexamer with V(O). PMID- 19289122 TI - Rab35 regulates neurite outgrowth and cell shape. AB - Recent studies have identified Rab35 in the endocytic pathway and as a regulator of cytokinesis; however its molecular mechanisms are currently unknown. Here, we find that Rab35 colocalizes with actin filaments and with Cdc42, Rac1 and RhoA, and that Rab35 can activate Cdc42 both in vivo and in vitro. We find activated Rab35 stimulates neurite outgrowth in PC12 and N1E-115 cells via a Cdc42 dependent pathway and that siRNA knockdown of Rab35 activity abolishes neurite outgrowth in these cell lines. We conclude that one function of Rab35 is to regulate Rho-family GTPases and that this role has consequences for neurite outgrowth. PMID- 19289123 TI - Interleukin-1beta is a positive regulator of TIARP/STAMP2 gene and protein expression in adipocytes in vitro. AB - The impact of interleukin (IL)-1beta on tumor necrosis factor alpha-induced adipose-related protein (TIARP)/six-transmembrane protein of prostate 2 (STAMP2) was determined in adipocytes. TIARP/STAMP2 mRNA synthesis was significantly stimulated by IL-1beta in a dose- and time-dependent fashion in 3T3-L1 adipocytes. Signaling studies suggested that janus kinase 2, nuclear factor kappaB, and p44/42 mitogen-activated protein kinase are involved in IL-1beta induced TIARP/STAMP2 mRNA expression. Furthermore, IL-1beta, TNFalpha, and IL-6 showed synergistic stimulatory effects on TIARP/STAMP2 gene expression. Moreover, both TIARP/STAMP2 mRNA synthesis and protein expression were induced by IL-1beta in fully differentiated human mesenchymal stem cell-derived adipocytes (hMSC-Ad). Taken together, TIARP/STAMP2 is highly upregulated in 3T3-L1 cells and hMSC-Ad by IL-1beta and might, therefore, modulate proinflammatory and insulin resistance inducing effects of IL-1beta. PMID- 19289124 TI - Intercellular exchange of proteins: the immune cell habit of sharing. AB - The recent recognition of new types of cell-cell communication pathways challenges classic theories of cell autonomy. Evidence of functional "proteome mixing" among interacting cells, particularly immune cells, supports the notion that no cell is an island, and that even these "unsplittable" units are actually non-autonomous. We summarize various mechanisms of intercellular transfer of proteins--trans-endocytosis, trogocytosis, exosomal transport, shuttle through nanotubes, and cell-contact-dependent intercellular transfer of intracellular proteins including oncogenic Ras. These phenomena suggest exciting new possibilities for proteome research, focusing on system-level proteomics that characterize cell contents and functions in the context of intercellular protein transfer. PMID- 19289125 TI - Ovarian aromatase and estrogens: a pivotal role for gonadal sex differentiation and sex change in fish. AB - The present review focuses on the roles of estrogens and aromatase (Cyp19a1a), the enzyme needed for their synthesis, in fish gonadal sex differentiation. Based on the recent literature, we extend the already well accepted hypothesis of an implication of estrogens and Cyp19a1a in ovarian differentiation to a broader hypothesis that would place estrogens and Cyp19a1a in a pivotal position to control not only ovarian, but also testicular differentiation, in both gonochoristic and hermaphrodite fish species. This working hypothesis states that cyp19a1a up-regulation is needed not only for triggering but also for maintaining ovarian differentiation and that cyp19a1a down-regulation is the only necessary step for inducing a testicular differentiation pathway. When considering arguments for and against, most of the information available for fish supports this hypothesis since either suppression of cyp19a1a gene expression, inhibition of Cyp19a1a enzymatic activity, or blockage of estrogen receptivity are invariably associated with masculinization. This is also consistent with reports on normal gonadal differentiation, and steroid-modulated masculinization with either androgens, aromatase inhibitors or estrogen receptor antagonists, temperature-induced masculinization and protogynous sex change in hermaphrodite species. Concerning the regulation of fish cyp19a1a during gonadal differentiation, the transcription factor foxl2 has been characterized as an ovarian specific upstream regulator of a cyp19a1a promoter that would co-activate cyp19a1a expression, along with some additional partners such as nr5a1 (sf1) or cAMP. In contrast, upstream factors potentially down-regulating cyp19a1a during testicular differentiation are still hypothetical, such as the dmrt1 gene, but their definitive characterization as testicular repressors of cyp19a1a would strongly strengthen the hypothesis that early testicular differentiation would need active repression of cyp19a1a expression. PMID- 19289126 TI - Mitochondria: from basic biology to cardiovascular disease. PMID- 19289127 TI - The structure of human extracellular copper-zinc superoxide dismutase at 1.7 A resolution: insights into heparin and collagen binding. AB - Extracellular superoxide dismutase (SOD3) is a homotetrameric copper- and zinc containing glycoprotein with affinity for heparin. The level of SOD3 is particularly high in blood vessel walls and in the lungs. The enzyme has multiple roles including protection of the lungs against hyperoxia and preservation of nitric oxide. The common mutation R213G, which reduces the heparin affinity of SOD3, is associated with increased risk of myocardial infarctions and stroke. We report the first crystal structure of human SOD3 at 1.7 A resolution. The overall subunit fold and the subunit-subunit interface of the SOD3 dimer are similar to the corresponding structures in Cu-Zn SOD (SOD1). The metal-binding sites are similar to those found in SOD1, but with Asn180 replacing Thr137 at the Cu binding site and a much shorter loop at the zinc-binding site. The dimers form a functional homotetramer that is fashioned through contacts between two extended loops on each subunit. The N- and C-terminal end regions required for tetramerisation and heparin binding, respectively, are highly flexible. Two grooves fashioned by the tetramer interface are suggestive as the probable sites for heparin and collagen binding. PMID- 19289128 TI - Dynamics of the ssDNA recognition by the RepA hexameric helicase of plasmid RSF1010: analyses using fluorescence stopped-flow intensity and anisotropy methods. AB - The kinetic mechanism of the single-stranded DNA (ssDNA) recognition by the RepA hexameric replicative helicase of the plasmid RSF1010 and the nature of formed intermediates, in the presence of the ATP nonhydrolyzable analog, beta,gamma imidoadenosine-5'-triphosphate (AMP-PNP), have been examined, using the fluorescence intensity and anisotropy stopped-flow and analytical ultracentrifugation methods. Association of the RepA hexamer with the ssDNA oligomers that engage the total DNA-binding site and exclusively the strong DNA binding subsite is a minimum four-step mechanism [formula: see text]. Extreme stability of the RepA hexamer precludes any disintegration of its structure, and the sequential character of the mechanism indicates that the enzyme exists in a predominantly single conformation prior to the association with the nucleic acid. Moreover, the hexameric helicase possesses a DNA-binding site located outside its cross channel. The reaction steps have dramatically different dynamics, with rate constants differing by 2-3 orders of magnitude. Such behavior indicates a very diverse nature of the observed transitions, which comprises binding steps and large conformational transitions of the helicase, including local opening of the hexameric structure. Steady-state fluorescence anisotropies of intermediates indicate that the entry of the DNA into the cross channel is initiated from the 5' end of the bound nucleic acid. The global structure of the tertiary complex RepA-ssDNA-AMP-PNP is very different from the structure of the binary complex RepA-AMP-PNP, indicating that, in equilibrium, the RepA hexamer-ssDNA-AMP-PNP complex exists as a mixture of partially open states. PMID- 19289129 TI - Two functions of the C-terminal domain of Escherichia coli Rob: mediating "sequestration-dispersal" as a novel off-on switch for regulating Rob's activity as a transcription activator and preventing degradation of Rob by Lon protease. AB - In Escherichia coli, Rob activates transcription of the SoxRS/MarA/Rob regulon. Previous work revealed that Rob resides in three to four immunostainable foci, that dipyridyl and bile salts are inducers of its activity, and that inducers bind to Rob's C-terminal domain (CTD). We propose that sequestration inactivates Rob by blocking its access to the transcriptional machinery and that inducers activate Rob by mediating its dispersal, allowing interaction with RNA polymerase. To test "sequestration-dispersal" as a new mechanism for regulating the activity of transcriptional activators, we fused Rob's CTD to SoxS and used indirect immunofluorescence microscopy to determine the effect of inducers on SoxS-Rob's cellular localization. Unlike native SoxS, which is uniformly distributed throughout the cell, SoxS-Rob is sequestered without an inducer, but is rapidly dispersed when cells are treated with an inducer. In this manner, Rob's CTD serves as an anti-sigma factor in regulating the co-sigma-factor-like activity of SoxS when fused to it. Rob's CTD also protects its N-terminus from Lon protease, since Lon's normally rapid degradation of SoxS is blocked in the chimera. Accordingly, Rob's CTD has novel regulatory properties that can be bestowed on another E. coli protein. PMID- 19289130 TI - Peroxisome proliferator-activated receptor alpha controls hepatic heme biosynthesis through ALAS1. AB - Heme is an essential prosthetic group of proteins involved in oxygen transport, energy metabolism and nitric oxide production. ALAS1 (5-aminolevulinate synthase) is the rate-limiting enzyme in heme synthesis in the liver and is highly regulated to adapt to the metabolic demand of the hepatocyte. In the present study, we describe human hepatic ALAS1 as a new direct target for the nuclear receptor peroxisome proliferator-activated receptor alpha (PPARalpha). In primary human hepatocytes and in HepG2 cells, PPARalpha agonists induced an increase in ALAS1 mRNA levels, which was abolished by PPARalpha silencing. These effects are mediated by two functional PPAR binding sites at positions -9 and -2.3 kb relative to the ALAS1 transcription start site. PPARalpha ligand treatment also up-regulated the mRNA levels of the genes ALAD (5-aminolevulinate dehydratase), UROS (uroporphyrinogen III synthase), UROD (uroporphyrinogen decarboxylase), CPOX (coproporphyrinogen oxidase) and PPOX (protoporphyrinogen oxidase) encoding for enzymes controlling further steps in heme biosynthesis. In HepG2 cells treated with PPARalpha agonists and in mouse liver upon fasting, the association of PPARalpha, its partner retinoid X receptor, PPARgamma co-activator 1alpha and activated RNA polymerase II with the transcription start site region of all six genes was increased, leading to higher levels of the metabolite heme. In conclusion, these data strongly support a role of PPARalpha in the regulation of human ALAS1 and of five additional genes of the pathway, consequently leading to increased heme synthesis. PMID- 19289131 TI - HIV-1 reverse transcriptase can simultaneously engage its DNA/RNA substrate at both DNA polymerase and RNase H active sites: implications for RNase H inhibition. AB - Reverse transcriptase of the human immunodeficiency virus possesses DNA polymerase and ribonuclease (RNase) H activities. Although the nucleic acid binding cleft separating these domains can accommodate structurally diverse duplexes, it is currently unknown whether regular DNA/RNA hybrids can simultaneously contact both active sites. In this study, we demonstrate that ligands capable of trapping the 3'-end of the primer at the polymerase active site affect the specificity of RNase H cleavage without altering the efficiency of the reaction. Experiments under single-turnover conditions reveal that complexes with a bound nucleotide substrate show specific RNase H cleavage at template position -18, while complexes with the pyrophosphate analogue foscarnet show a specific cut at position -19. This pattern is indicative of post translocated and pre-translocated conformations. The data are inconsistent with models postulating that the substrate toggles between both active sites, such that the primer 3'-terminus is disengaged from the polymerase active site when the template is in contact with the RNase H active site. In contrast, our findings provide strong evidence to suggest that the nucleic acid substrate can engage both active sites at the same time. As a consequence, the bound and intact DNA/RNA hybrid can restrict access of RNase H active site inhibitors. We have mapped the binding site of the recently discovered inhibitor beta-thujaplicinol between the RNase H active site and Y501 of the RNase H primer grip, and have shown that the inhibitor is unable to bind to a preformed reverse transcriptase DNA/RNA complex. In conclusion, the bound nucleic acid substrate and in turn, active DNA synthesis can represent an obstacle to RNase H inhibition with compounds that bind to the RNase H active site. PMID- 19289132 TI - Plant physiology in theory and practice: an analysis of the WBE model for vascular plants. AB - The theoretical model of West, Brown and Enquist (hereafter WBE) proposed the fractal geometry of the transport system as the origin of the allometric scaling laws observed in nature. The WBE model has either been criticized for some restrictive and biologically unrealistic constraints or its reliability debated on the evidence of empirical tests. In this work, we revised the structure of the WBE model for vascular plants, highlighting some critical assumptions and simplifications and discuss them with regard to empirical evidence from plant anatomy and physiology. We conclude that the WBE model had the distinct merit of shedding light on some important features such as conduit tapering. Nonetheless, it is over-simplistic and a revised model would be desirable with an ontogenetic perspective that takes some important phenomena into account, such as the transformation of the inner sapwood into heartwood and the effect of hydraulic constraints in limiting the growth in height. PMID- 19289133 TI - Timely identification of optimal control strategies for emerging infectious diseases. AB - BACKGROUND: Health authorities must rely on quarantine, isolation, and other non pharmaceutical interventions to contain outbreaks of newly emerging human diseases. METHODS: We modeled a generic disease caused by a pathogen apparently transmitted by close interpersonal contact, but about which little else is known. In our model, people may be infectious while incubating or during their prodrome or acute illness. We derived an expression for Re, the reproduction number, took its partial derivatives with respect to control parameters, and encoded these analytical results in a user-friendly Mathematica notebook. With biological parameters for SARS estimated from the initial case series in Hong Kong and infection rates from hospitalizations in Singapore, we determined Re's sensitivity to control parameters. RESULTS: Stage-specific infection rate estimates from cases hospitalized before quarantine began exceed those from the entire outbreak, but are qualitatively similar: infectiousness was negligible until symptom onset, and increased 10-fold from prodrome to acute illness. Given such information, authorities might instead have emphasized a strategy whose efficiency more than compensates for any possible reduction in efficacy. CONCLUSIONS: In future outbreaks of new human diseases transmitted via close interpersonal contact, it should be possible to identify the optimal intervention early enough to facilitate effective decision-making. PMID- 19289134 TI - Accelerating invasion rates result from the evolution of density-dependent dispersal. AB - Evolutionary processes play an important role in shaping the dynamics of range expansions, and selection on dispersal propensity has been demonstrated to accelerate rates of advance. Previous theory has considered only the evolution of unconditional dispersal rates, but dispersal is often more complex. For example, many species emigrate in response to crowding. Here, we use an individual-based model to investigate the evolution of density dependent dispersal into empty habitat, such as during an invasion. The landscape is represented as a lattice and dispersal between populations follows a stepping-stone pattern. Individuals carry three 'genes' that determine their dispersal strategy when experiencing different population densities. For a stationary range we obtain results consistent with previous theoretical studies: few individuals emigrate from patches that are below equilibrium density. However, during the range expansion of a previously stationary population, we observe evolution towards dispersal strategies where considerable emigration occurs well below equilibrium density. This is true even for moderate costs to dispersal, and always results in accelerating rates of range expansion. Importantly, the evolution we observe at an expanding front depends upon fitness integrated over several generations and cannot be predicted by a consideration of lifetime reproductive success alone. We argue that a better understanding of the role of density dependent dispersal, and its evolution, in driving population dynamics is required especially within the context of range expansions. PMID- 19289135 TI - Exercise training improves cutaneous microvascular endothelial function in post surgical varicose vein patients. AB - This study investigated the effects of exercise training on cutaneous microvascular function in post-surgical varicose vein patients. Sixteen post surgical (4-5 weeks) varicose vein patients were randomised to a treadmill walking exercise group or a non-exercise control group. The exercise group trained twice weekly for 8 weeks. Changes in cutaneous microvascular function of the gaiter area were assessed using laser Doppler flowmetry combined with incremental-dose iontophoretic administration of acetylcholine chloride (ACh) and sodium nitroprusside (SNP) in both supine and standing positions. At 8 weeks, peak flux responses to ACh in the supine position were increased in the exercise group (44+/-30 to 62+/-33 PU; P=0.03) with a similar trend in the standing position (37+/-27 to 74+/-31 PU; P=0.08). There were no such changes in the control group (P>0.05). Additionally, peak flux responses to SNP were unchanged in both groups and body positions (P>0.05). The results suggest that moderate intensity lower-limb exercise training improves microvascular endothelial vasodilator function in post-surgical varicose vein patients. PMID- 19289136 TI - C-338A polymorphism of the endothelin-converting enzyme-1 gene and the susceptibility to carotid atherosclerosis. AB - Several lines of evidence supported the involvement of ECE-1 in the development of atherosclerosis disease. We investigated the C-338A polymorphism of the ECE-1b gene in carotid atherosclerosis. A cohort of 518 Chinese carotid atherosclerosis patients and 518 controls matched by age and sex were examined for the ECE-1b C 338A polymorphism. The ECE-1b-338 A allele was significantly frequent in carotid atherosclerosis patients than in controls (chi(2)=9.167; P=0.002). A alleles had a 26% increased risk of carotid atherosclerosis compared with C alleles (OR=1.26; 95% CI=1.05-1.52). In stratified analyses, the A allele carrying genotypes was significantly associated with an increased risk of carotid atherosclerosis in females (adjusted OR=1.57; 95% CI=1.14-2.23) and a subject with age >or=64 years (adjusted OR=1.72; 95% CI=1.23-1.41). Our results suggested that ECE-1b-338 A allele carrying genotypes might be associated with increased risk of carotid atherosclerosis in the Chinese population. PMID- 19289138 TI - The food contaminant deoxynivalenol, decreases intestinal barrier permeability and reduces claudin expression. AB - 'The gastrointestinal tract represents the first barrier against food contaminants as well as the first target for these toxicants. Deoxynivalenol (DON) is a mycotoxin that commonly contaminates cereals and causes various toxicological effects. Through consumption of contaminated cereals and cereal products, human and pigs are exposed to this mycotoxin. Using in vitro, ex vivo and in vivo approaches, we investigated the effects of DON on the intestinal epithelium. We demonstrated that, in intestinal epithelial cell lines from porcine (IPEC-1) or human (Caco-2) origin, DON decreases trans-epithelial electrical resistance (TEER) and increases in a time and dose-dependent manner the paracellular permeability to 4 kDa dextran and to pathogenic Escherichia coli across intestinal cell monolayers. In pig explants treated with DON, we also observed an increased permeability of intestinal tissue. These alterations of barrier function were associated with a specific reduction in the expression of claudins, which was also seen in vivo in the jejunum of piglets exposed to DON contaminated feed. In conclusion, DON alters claudin expression and decreases the barrier function of the intestinal epithelium. Considering that high levels of DON may be present in food or feed, consumption of DON-contaminated food/feed may induce intestinal damage and has consequences for human and animal health. PMID- 19289137 TI - Excitatory and inhibitory synaptic transmission is differentially influenced by two ortho-substituted polychlorinated biphenyls in the hippocampal slice preparation. AB - Exposure to polychlorinated biphenyls impairs cognition and behavior in children. Two environmental PCBs 2,2',3,3',4,4',5-heptachlorobiphenyl (PCB170) and 2,2',3,5',6-pentachlorobiphenyl (PCB95) were examined in vitro for influences on synaptic transmission in rat hippocampal slices. Field excitatory postsynaptic potentials (fEPSPs) were recorded in the CA1 region using a multi-electrode array. Perfusion with PCB170 (10 nM) had no effect on fEPSP slope relative to baseline period, whereas (100 nM) initially enhanced then depressed fEPSP slope. Perfusion of PCB95 (10 or 100 nM) persistently enhanced fEPSP slope >200%, an effect that could be inhibited by dantrolene, a drug that attenuates ryanodine receptor signaling. Perfusion with picrotoxin (PTX) to block GABA neurotransmission resulted in a modest increase in fEPSP slope, whereas PTX+PCB170 (1-100 nM) persistently enhanced fEPSP slope in a dose dependent manner. fEPSP slope reached >250% of baseline period in the presence of PTX+100 nM PCB170, conditions that evoked marked epileptiform after-potential discharges. PCB95 and PCB170 were found to differentially influence the Ca(2+)-dependence of [(3)H]ryanodine-binding to hippocampal ryanodine receptors. Non-coplanar PCB congeners can differentially alter neurotransmission in a manner suggesting they can elicit imbalances between inhibitory and excitatory circuits within the hippocampus. Differential sensitization of ryanodine receptors by Ca(2+) appears to mediate, at least in part, hippocampal excitotoxicity by non-coplanar PCBs. PMID- 19289139 TI - Three-dimensional shape from second-order orientation flows. AB - In images of textured surfaces, orientation flows formed by perspective convergence invariably convey 3D shape. We show that orientation flows formed by contrast-modulated (CM) and illusory contours (IC) convey 3D shape, and that both stimulus types induce 3D shape aftereffects on CM and IC test stimuli. Adaptation to luminance-modulated (LM) orientation flows induce robust 3D shape aftereffects on CM and IC tests, however, aftereffects using CM/IC adapting stimuli on LM tests were substantially weaker. These results can be explained by the adaptation of 3D shape-selective neurons that invariantly extract first- and second-order orientation flows from striate and extra-striate signals, which receive stronger input from neurons selective for first-order orientation flows. PMID- 19289140 TI - Task-irrelevant stimulus salience affects visual search. AB - The relative contributions of stimulus salience and task-related goals in guiding attention remain an issue of debate. Several studies have demonstrated that top down factors play an important role, as they often override capture by salient irrelevant objects. However, Yantis and Egeth [Yantis, S., & Egeth, H. E. (1999). On the distinction between visual salience and stimulus-driven attentional capture. Journal of Experimental Psychology: Human Perception and Performance, 25, 661-676.] have made the more radical claim that salience plays no role in visual search unless the observer adopts an attentional set for singletons or "singleton-detection mode". We reexamine their claim while disentangling effects of stimulus salience from effects of attentional set and inter-trial repetition. The results show that stimulus salience guides attention even when salience is task irrelevant. PMID- 19289141 TI - Liver cancer mortality among male prison inmates in Texas, 1992-2003. AB - OBJECTIVES: Prevalence estimates for several liver cancer risk factors-hepatitis C, hepatitis B, and history of alcohol abuse-are substantially higher in U.S. prison populations than in the general population. However, liver cancer mortality data from these populations are lacking. The primary aims of this study were to examine trends in liver cancer mortality rates from 1992 to 2003 among male prisoners in the Texas Department of Criminal Justice (TDCJ) and to compare these rates to general population rates. METHODS: TDCJ data on male decedents (N=4026) were linked with Texas Vital Statistics multiple-cause-of-death data. Crude average annual liver cancer death rates, average annual percent changes, and standardized mortality ratios were estimated. RESULTS: Crude liver cancer death rates increased by an average annual 6.1% among male prisoners, which was considerably higher than the average annual percent change among similarly aged males in Texas (2.0%) and the U.S. (2.9%). The number of liver cancer deaths among male prisoners was 4.7 (4.0-5.6) and 6.3 (5.3-7.5) times higher than the expected number of deaths estimated using age-specific rates from these reference populations. CONCLUSIONS: From 1992 to 2003, liver cancer death rates and rate increases were elevated among Texas male prisoners. Findings support previous recommendations for targeted prevention, screening, and treatment of liver cancer risk factors in prison populations. PMID- 19289142 TI - From pills to programs: lessons from medicine for developing effective lifestyle interventions. AB - OBJECTIVE: To propose a scheme for comprehensive development and evaluation of lifestyle interventions. METHODS: We adapted the four-phase system used in drug development, the engine of progress in medicine for decades, to construct a system for developing lifestyle intervention programs. RESULTS: Phase I: The intervention is constructed and tested with a small number of individuals. Acceptability and feasibility are assessed. Evaluation is primarily qualitative. Phase II: Effectiveness on intermediate endpoints (e.g. behavior) is tested in a real field setting, with a limited number of individuals, using a before-and after design. An iterative process of testing and refinement may be necessary. Phase III: The effectiveness of the intervention on health-related outcomes is tested, using, where possible, a randomized design. Phase IV: Large-scale implementation and penetration are assessed in other populations. Process variables and local and national health indicators are studied. The development and evaluation of our hygiene intervention, which took place in Jerusalem from 1999 to 2001, is presented as a case study. CONCLUSIONS: Adaptation of the phased system of drug development to lifestyle interventions is a conceptually simple approach to building effective, sustainable programs for community-based public health. PMID- 19289143 TI - The effects of GLP-1 infusion in the hepatic portal region on food intake. AB - A wide range of evidence points to a role for GLP-1 to regulate food intake. Anorectic effects of GLP-1 are most apparent when the peptide is administered directly into the central nervous system (CNS), but suppression of food intake has also been noted in some cases with peripheral administration. It is unclear which GLP-1 receptor (GLP-1r) population mediates the effects of plasma GLP-1, although direct actions to activate CNS neurons have been demonstrated. More recently several groups have demonstrated that GLP-1 can activate peripheral nerves in the hepatic portal vein to regulate glucose metabolism. To test the hypothesis that GLP-1 receptors on nerve terminals in the hepatic portal affect feeding behavior, we compared the effects of direct infusions into hepatic portal and jugular veins in rats. Jugular GLP-1 decreased food intake at doses as low as 10 microg from 0.5-4 h into the dark cycle, whereas portal GLP-1 decreased food intake only at the highest dose tested (100 microg). The blockade of endogenous GLP-1 action before or during eating by infusing dH-Ex, GLP-1 receptor antagonist, into either jugular or portal vein did not cause any change in food intake during either the dark or light cycles. Taken together, these data suggest that while peripheral GLP-1 may be involved in the regulation of food intake, the key GLP-1 receptors are unlikely to be those associated with vagal afferent nerves innervating the hepatic portal vein. PMID- 19289144 TI - Autoregulation of human relaxin-2 gene expression critically involves relaxin and glucocorticoid receptor binding to glucocorticoid response half-sites in the relaxin-2 promoter. AB - Relaxin peptides act in brain, reproductive and cardiovascular systems, kidneys, and connective tissue through different G protein-coupled receptors. We reported that human relaxin-2 and porcine relaxin are both agonists at the human glucocorticoid receptor (GR). Here, we investigated the possible auto-regulation of relaxin-2 gene expression via recently discovered GR-binding sites in the relaxin-2 promoter. We found that porcine relaxin increased the secretion of human relaxin-like immunoreactivity in HeLa and THP-1 cells. Silencing of GR gene expression completely abolished this effect whereas transfection of wild-type GR into naturally GR-devoid HT-29 cells established relaxin sensitivity. Relaxin was shown to stimulate CAT expression driven by different deletion constructs of the 5'-flanking region of the relaxin-2 promoter. In chromatin immunoprecipitation assays, we detected both GR and relaxin binding to the relaxin-2 promoter. Gel shift assays indicated binding of relaxin-activated GR to half-GREs located between 160 and 200 bp upstream of transcription start but not to the GRE at -900 bp. Relaxin bound to human GR and displaced established GR agonists. Immunofluorescence experiments visualized nuclear co-localization of relaxin and GR in response to relaxin. In conclusion, we have identified a positive auto regulatory loop of human relaxin-2 expression which involves GR and relaxin/GR binding to half-GREs in the relaxin-2 promoter. PMID- 19289145 TI - Effects of galanin-monoaminergic interactions on vasopressin secretion in rat neurohypophyseal cell cultures. AB - The effects of dopamine (DA), serotonin (5-HT), histamine (HA), adrenaline (ADR), noradrenaline (NADR) and K(+) administration on vasopressin (VP) secretion were studied in 13-14-day cultures of rat neurohypophyseal (NH) cells, and it was examined whether galanin (GAL) can modify the VP release enhancement induced by these monoaminergic compounds. An enzymatic dissociation technique was used to make the rat NH cell cultures. The VP contents of the supernatants of 14-day cultures were determined by radioimmunoassay. Following the administration of 10( 6) M GAL, the VP secretion into the supernatant media decreased. DA, 5-HT, ADR or NADR treatment increased the VP level substantially, while the enhancing effect of HA was more moderate. GAL administration before DA, ADR and NADR treatment prevented the VP concentration increase induced by DA, ADR or NADR. Preincubation with GAL reduced the 5-HT- or HA-induced VP level increases; the VP concentrations of the supernatant media remained above the control level. The GAL blocking effect was prevented by previous treatment with the GAL receptor antagonist galantid (M15). GAL had no effect on the VP level increase induced by K(+), which causes a non-specific hormone secretion. The results indicate that the changes in VP secretion induced by the monoaminergic system can be directly influenced by the GAL-ergic system. The interactions between the monoaminergic and GAL-ergic systems regarding VP secretion occur at the level of the posterior pituitary. PMID- 19289146 TI - Ghrelin inhibits apoptosis induced by high glucose and sodium palmitate in adult rat cardiomyocytes through the PI3K-Akt signaling pathway. AB - OBJECTIVE: Ghrelin is a gastric acyl-peptide that has been identified as an endogenous ligand for the growth hormone secretagogue receptor. It has been reported to have cardioprotective activities independent of growth hormone release. We investigated the effect of ghrelin on apoptosis induced by high glucose and sodium palmitate and the mechanisms underlying the cardioprotective activities of ghrelin. RESEARCH DESIGN AND METHODS: Cardiomyocytes were isolated from hearts of adult rats and cultured in serum-free MEM. High glucose (30 mM) or sodium palmitate (0.5 mM) were used to induce apoptosis. Apoptosis was detected using an annexin V-FITC/PI binding assay and a caspase 3 activity assay. Reactive oxygen species were detected using a DCFH-DA fluorescent probe. Phospho-Akt, phospho-ERK, and NF kappaB levels were determined using ELISA. The transcription of genes was analyzed using real-time PCR. RESULTS: Ghrelin can inhibit apoptosis induced by oxidative stress in cardiomyocytes from adult rats through the activation of the PI3K-Akt signaling pathway. In addition, ghrelin does not decrease intracellular oxidative stress. Activation of the MEK-ERK1/2 signaling pathway has no influence on the inhibition of apoptosis. Finally, ghrelin activates NF kappaB and subsequently increases the transcription of survival genes such as Bcl-2, Bcl-xL, c-iap, and c-fos. CONCLUSION: Our research provides evidence that ghrelin may act as a survival factor under oxidative stress in cardiomyocytes. This may provide a clue for therapy for myocardial disease in diabetes mellitus. PMID- 19289147 TI - Mitophagy. AB - Concurrent mitochondrial elimination and autophagy in many systems has led to the proposal that autophagy is the main mechanism of mitochondrial turnover during development and under pathological conditions. The term mitophagy was coined to describe the selective removal of mitochondria by autophagy but the process itself is still contentious. Three questions are being debated: 1) Is there a specific removal of mitochondria by autophagy or is it non-selective or inadvertent? 2) What are the signals that drive this process? 3) Does removal of mitochondria increase or decrease cell viability? There is a mounting evidence for specific signals in/on mitochondria that drive mitochondrial removal from cells by autophagy. The process itself may be both selective and non-selective. In yeast, surprisingly, mitochondrial elimination occurs more by microautophagy (intracellular pinocytosis by the vacuolar membrane) than macroautophagy (initiated by stand-alone nascent double membrane structures known as autophagosomes). In mammalian cells, macroautophagy seems most prevalent though tools to study microautophagy are not well developed. Whilst lack of mitophagy seems to be deleterious, understanding the interplay between autophagy, mitochondrial performance, and cell pathology is a much-needed area of research. PMID- 19289148 TI - Encapsulation of the synthetic retinoids Am80 and LE540 into polymeric micelles and the retinoids' release control. AB - The objective of this study was to encapsulate two synthetic retinoids Am80 and LE540 into polymeric micelles and to control the retinoids' release rate in vitro. Highly efficient encapsulation yields of these retinoids were obtained for micelles forming from PEG-poly(benzyl aspartate) block copolymers in the wide range of the benzyl substitution degree. The in vitro release examination for LE540 indicated very stable encapsulation of this retinoid owing to its strongly hydrophobic nature. On the other hand, Am80 exhibited a rapid release in Dulbecco's phosphate buffer saline. An addition of a hydrophobic alkyl amine in the Am80-encapsulation process successfully led to significant retardation of the Am80 release rate. A mechanism of the retardation was considered an increase of Am80 hydrophobicity due to an ion-pairing with the alkyl amine. This paper is the first report on release control in the polymeric micelle carrier system through the ion-pairing between an encapsulated drug and an additive. PMID- 19289149 TI - Glutathione peroxidases in different stages of carcinogenesis. AB - Cancer cells produce high amounts of reactive oxygen species (ROS) and evade apoptosis. Hydroperoxides support proliferation, invasion, migration and angiogenesis, but at higher levels induce apoptosis, thus being pro- and anti carcinogenic. Accordingly, glutathione peroxidases (GPxs) regulating hydroperoxide levels might have dual roles too. GPx1, clearly an antioxidant enzyme, is down-regulated in many cancer cells. Its main role would be prevention of cancer initiation by ROS-mediated DNA damage. GPx2 is up-regulated in cancer cells. GPx1/GPx2 double knockout mice develop colitis and intestinal cancer. However, GPx2 knockdown cancer cells grow better in vitro and in vivo probably reflecting the physiological role of GPx2 in intestinal mucosa homeostasis. GPx2 counteracts COX-2 expression and PGE(2) production, which explains its potential to inhibit migration and invasion of cultured cancer cells. Overexpression of GPx3 inhibits tumor growth and metastasis. GPx4 is decreased in cancer tissues. GPx4-overexpressing cancer cells have low COX-2 activity and tumors derived therefrom are smaller than from control cells and do not metastasize. Collectively, GPxs prevent cancer initiation by removing hydroperoxides. GPx4 inhibits but GPx2 supports growth of established tumors. Metastasis, but also apoptosis, is inhibited by all GPxs. GPx-mediated regulation of COX/LOX activities may be relevant to early stages of inflammation-mediated carcinogenesis. PMID- 19289151 TI - Studies on substrate specificity and activity regulating factors of trehalose-6 phosphate synthase of Saccharomyces cerevisiae. AB - Purified trehalose-6-phosphate synthase (TPS) of Saccharomyces cerevisiae was effective over a wide range of substrates, although differing with regard to their relative activity. Polyanions heparin and chondroitin sulfate were seen to stimulate TPS activity, particularly when a pyrimidine glucose nucleotide like UDPG was used, rather than a purine glucose nucleotide like GDPG. A high V(max) and a low K(m) value of UDPG show its greater affinity with TPS than GDPG or TDPG. Among the glucosyl acceptors TPS showed maximum activity with G-6-P which was followed by M-6-P and F-6-P. Effect of heparin was also extended to the purification of TPS activity, as it helped to retain both stability and activity of the final purified enzyme. Metal co-factors, specifically MnCl(2) and ZnCl(2) acted as stimulators, while enzyme inhibitors had very little effect on TPS activity. Metal chelators like CDTA, EGTA stimulated enzyme activity by chelation of metal inhibitors. Temperature and pH optima of the purified enzyme were determined to be 40 degrees C and pH 8.5 respectively. Enzyme activity was stable at 0-40 degrees C and at alkaline pH. PMID- 19289150 TI - Regulation of integrin activity and signalling. AB - The ability of cells to attach to each other and to the extracellular matrix is of pivotal significance for the formation of functional organs and for the distribution of cells in the body. Several molecular families of proteins are involved in adhesion, and recent work has substantially improved our understanding of their structures and functions. Also, these molecules are now being targeted in the fight against disease. However, less is known about how their activity is regulated. It is apparent that among the different classes of adhesion molecules, the integrin family of adhesion receptors is unique in the sense that they constitute a large group of widely distributed receptors, they are unusually complex and most importantly their activities are strictly regulated from the inside of the cell. The activity regulation is achieved by a complex interplay of cytoskeletal proteins, protein kinases, phosphatases, small G proteins and adaptor proteins. Obviously, we are only in the beginning of our understanding of how the integrins function, but already now fascinating details have become apparent. Here, we describe recent progress in the field, concentrating mainly on mechanistical and structural studies of integrin regulation. Due to the large number of articles dealing with integrins, we focus on what we think are the most exciting and rewarding directions of contemporary research on cell adhesion and integrins. PMID- 19289152 TI - From heterochromatin islands to the NAD World: a hierarchical view of aging through the functions of mammalian Sirt1 and systemic NAD biosynthesis. AB - For the past couple of decades, aging science has been rapidly evolving, and powerful genetic tools have identified a variety of evolutionarily conserved regulators and signaling pathways for the control of aging and longevity in model organisms. Nonetheless, a big challenge still remains to construct a comprehensive concept that could integrate many distinct layers of biological events into a systemic, hierarchical view of aging. The "heterochromatin island" hypothesis was originally proposed 10 years ago to explain deterministic and stochastic aspects of cellular and organismal aging, which drove the author to the study of evolutionarily conserved Sir2 proteins. Since a surprising discovery of their NAD-dependent deacetylase activity, Sir2 proteins, now called "sirtuins," have been emerging as a critical epigenetic regulator for aging. In this review, I will follow the process of conceptual development from the heterochromatin island hypothesis to a novel, comprehensive concept of a systemic regulatory network for mammalian aging, named "NAD World," summarizing recent studies on the mammalian NAD-dependent deacetylase Sirt1 and nicotinamide phosphoribosyltransferase (Nampt)-mediated systemic NAD biosynthesis. This new concept of the NAD World provides critical insights into a systemic regulatory mechanism that fundamentally connects metabolism and aging and also conveys the ideas of functional hierarchy and frailty for the regulation of aging in mammals. PMID- 19289154 TI - Increase of intracellular Ca2+ by P2X and P2Y receptor-subtypes in cultured cortical astroglia of the rat. AB - Astrocytes express purinergic receptors that are involved in glial-neuronal cell communication. Experiments were conducted to characterize the expression of functional P2X/P2Y nucleotide receptors in glial cells of mixed cortical cell cultures of the rat. The vast majority of these cells was immunopositive for glial fibrillary acidic protein (GFAP) and was considered therefore astrocyte like; for the sake of simplicity they were termed "astroglia" throughout. Astroglia expressed predominantly P2X(4,6,7) as well as P2Y(1,2) receptor subtypes. Less intensive immunostaining was also found for P2X(5) and P2Y(4,6,13,14) receptors. Pressure application of ATP and a range of agonists selective for certain P2X or P2Y receptor-subtypes caused a concentration dependent increase of intracellular Ca(2+) ([Ca(2+)](i)). Of the agonists tested, only the P2X(1,3) receptor-selective alpha,beta-methylene ATP was ineffective. Experiments with Ca(2+)-free solution and cyclopiazonic acid, an inhibitor of the endoplasmic Ca(2+)-ATPase, indicated that the [Ca(2+)](i) response to most nucleotides, except for ATP and 2',3'-O-(benzoyl-4-benzoyl)-ATP, was due primarily to the release of Ca(2+) from intracellular stores. A Gprotein-mediated release of Ca(2+) is the typical signaling mechanism of various P2Y receptor subtypes, whose presence was confirmed also by cross-desensitization experiments and by using selective antagonists. Thus, our results provide direct evidence that astroglia in mixed cortical cell cultures express functional P2Y (P2Y(1,2,6,14) and probably also P2Y(4)) receptors. Several unidentified P2X receptors, including P2X(7), may also be present, although they appear to only moderately participate in the regulation of [Ca(2+)](i). The rise of [Ca(2+)](i) is due in this case to the transmembrane flux of Ca(2+) via the P2X receptor channel. In conclusion, P2Y rather than P2X receptor-subtypes are involved in modulating [Ca(2+)](i) of cultured astroglia and thereby may play an important role in cell-to-cell signaling. PMID- 19289153 TI - Molecular mechanisms by which selenoproteins affect cancer risk and progression. AB - Selenoproteins comprise a unique class of proteins that contain selenium in the form of selenocysteine. Several selenoproteins have been implicated in the risk or development of cancers in humans by genetic data. These include Selenoprotein P, 3 members of the glutathione peroxidase family of anti-oxidant enzymes and Sep15. At-risk alleles in the germline indicate a likely role in determining susceptibility to cancer, while loss of heterozygosity or chromosomal epigenetic silencing indicate that the reduction in the levels of the corresponding proteins contribute to malignant progression. Lower levels of these proteins are likely to be detrimental due to the resulting cellular stress and perturbations in important regulatory signaling pathways. The genetic data indicating the involvement of these selenoproteins in cancer etiology are discussed, as are the possible mechanisms by which these genes might promote carcinogenesis. PMID- 19289155 TI - EphB2 and EphA4 receptors regulate formation of the principal inter-hemispheric tracts of the mammalian forebrain. AB - Previously, we have demonstrated that EphB2 activity is required for proper development of the posterior branch of the anterior commissure (ACpp) within the mammalian forebrain. In the present study, using magnetic resonance imaging (MRI), immunohistochemistry, and in vivo stereotactic fluorescence tracing of EphB2, B3, A4 and combinatorial Eph receptor mutants, we have developed a detailed three-dimensional model of how EphB-class receptors interact to regulate commissural formation within the forebrain. The results demonstrate that EphB2 and EphA4 each regulate distinct aspects of axon guidance within the ACpp. Specifically, while EphB2 is required to retard ACpp axons from projecting aberrantly into the ventral forebrain, EphA4 is required to restrict axons from entering the anterior branch of the anterior commissure (ACpa). Together, EphB2 and EphA4 act synergistically to prevent a subpopulation of axons within the anterior branch of the AC from mis-projecting caudally. Analysis of EphA4 null mice using high resolution MRI reveals for the first time that, in addition to errors in midline guidance, loss of EphA4 results in aberrant lateral and ventral displacement of the ACpa tract. In addition, tracing studies in alpha-chimerin null mice reveal that EphA4-mediated effects are not regulated through this pathway. Taken together, the results demonstrate that each of the principal guidance decisions within both anterior and posterior tracts of the anterior commissure can be accounted for by the individual and combinatorial actions of EphB2/A4 receptors. PMID- 19289156 TI - Antipsychotics affect multiple calcium calmodulin dependent proteins. AB - Calcineurin is a calmodulin (CaM) dependent protein phosphatase recently found to be altered in the brains of patients suffering from schizophrenia and by repeated antipsychotic treatment in rats. Some data suggest, however, that antipsychotics and schizophrenia may have a more widespread effect on the CaM signaling axis than calcineurin alone. In the current study, the effects of selected psychoactive drugs were investigated using Western blotting, in situ hybridization and immunocytochemistry to determine if they target CaM, calmodulin dependent protein kinases (CaMK) or calcineurin. Results indicated that repeated treatment with haloperidol, clozapine or risperidone increased CaM protein and CaMII mRNA levels but decreased calmodulin-dependent protein kinase IIalpha (CaMKIIalpha) IV (CaMKIV), kinase alpha (CaMKKalpha), kinase beta (CaMKKbeta) and calcineurin protein levels in the striatum of Sprague-Dawley rats (Rattus Norvegicus). Closer examination of CaMKIV, CaMKKalpha and CaMKKbeta revealed that the observed decreases in protein levels were short-lived following antipsychotic treatment and reversed (i.e. upregulated) 24 h post-treatment similar to what was previously reported for calcineurin. The D(2)/D(3)dopamine receptor antagonist raclopride mimicked the decreases in CaMKIV, CaMKKalpha, CaMKKbeta and calcineurin observed following antipsychotic treatment whereas increases in these proteins were observed in an amphetamine model of the positive symptoms of schizophrenia. Mood stabilizers such as lithium and valproic acid or the antidepressant fluoxetine had no effect on CaMKIV, CaMKKalpha, CaMKKbeta and calcineurin with the exception of an increase in CaMKKbeta following lithium treatment. The results collectively suggest that antipsychotic specifically target several proteins associated with CaM signaling. PMID- 19289157 TI - Dihydropyridine block of voltage-dependent K+ currents in rat dorsal root ganglion neurons. AB - The dihydropyridines nifedipine, nimodipine and Bay K 8644 are widely used as pharmacological tools to assess the contribution of L-type voltage-gated Ca(2+) channels to a variety of neuronal processes including synaptic transmission, excitability and second messenger signaling. These compounds are still used in neuronal preparations despite evidence from cardiac tissue and heterologous expression systems that they block several voltage-dependent K(+) (Kv) channels. Both because these compounds have been used to assess the relative contribution of L-type Ca(2+) channels to several different processes in dorsal root ganglion (DRG) neurons and because a relatively wide variety of Kv channels present in other neuronal populations is present in DRG neurons, we determined the extent to which dihydropyridines block Kv currents in these neurons. Standard whole cell patch clamp techniques were used to study acutely disassociated adult rat DRG neurons. All three dihydropyridines tested blocked Kv currents in DRG neurons; IC(50) values (concentration resulting in an inhibition that is 50% of maximum) for nifedipine and nimodipine-induced block of sustained Kv currents were 14.5 and 6.6 microM, respectively. The magnitude of sustained current block was 44+/ 1.6%, 60+/-2%, and 56+/-2.9% with 10 microM nifedipine, nimodipine and Bay K 8644, respectively. Current block was occluded by neither 4-aminopyridine (5 mM) nor tetraethylammonium (135 mM). Dihydropyridine-induced block of Kv currents was not associated with a shift in the voltage-dependence of current activation or inactivation, the recovery from inactivation, or voltage dependent block. However, there was a small use-dependence to the dihydropyridine-induced block. Our results suggest that several types of Kv channels in DRG neurons are blocked by mechanisms distinct from those underlying block of Kv channels in cardiac myocytes. Importantly, our results suggest that if investigators wish to explore the contribution of L-type Ca(2+) channels to neuronal function, they should consider alternative strategies for the manipulation of these channels than the use of dihydropyridines. PMID- 19289158 TI - Biologically active components of a Papua New Guinea analgesic and anti inflammatory lichen preparation. AB - A traditional preparation of Parmotrema saccatilobum (Taylor) Hale (Family: Parmeliaceae) is being considered for inclusion into the PNG national drug formulary by the Ministry of Health Taskforce on Traditional Medicines. The lichen preparation is traditionally used in the Milne Bay province of Papua New Guinea for analgesic and anti-inflammatory activities. A hexane extract of P. saccatilobum yielded the principle components atranorin and chloroatranorin. Atranorin and chloroatranorin were tested in a COX-1 and -2 enzyme inhibition assay, which showed that atranorin inhibited COX-1 in a dose dependent manner and suggests partial inhibition by atranorin and chloroatranorin of COX-2 and COX-1, respectively. PMID- 19289159 TI - Metabolic profiles of three related Salvia species. AB - Salvia miltiorrhiza Bunge is one of the most important and popular plant of the traditional Chinese medicine (TCM), but Salvia castanea Diels f. tomentosa Stib and Salvia miltiorrhiza Bunge f. alb have also been reported to have the same therapeutic effects as S. miltiorrhiza. To better distinguish between these species, the phytochemical profiles of three Salvia species were investigated by liquid chromatography. All the Salvia species were good sources of tanshinones, with the contents of phenolics being high in S. miltiorrhiza and S. miltiorrhiza f. alb, but not in S. castanea Diels f. tomentosa Stib. These results pave the way for a better phytotherapy exploitation of these plants. PMID- 19289160 TI - Identification of transcription factors predominantly expressed in soybean flowers and characterization of GmSEP1 encoding a SEPALLATA1-like protein. AB - By microarray analysis and real-time RT-PCR, we identified 28 soybean flower enriched transcription factors such as MADS-box proteins, zinc finger proteins, and MYB proteins. Among them, one MADS-box protein GmSEP1 was chosen for further analysis. GmSEP1 contains 8 exons and 7 introns, showing similar exon-intron structure with Arabidopsis SEP genes. Phylogenetic analysis also suggested that GmSEP1 belonged to AGL2/SEP subfamily and likely a soybean orthologue of Arabidopsis SEP1. Subcellular localization assay suggested that GmSEP1 was localized in nucleus. Semi-quantitative RT-PCR showed that GmSEP1 was predominantly expressed in reproductive organs including petal, sepal, stamen, carpel and seed. By real-time RT-PCR, we found that GmSEP1 was expressed with a low level in stamens of soybean mutant NJS-10Hfs in which some stamens are converted to petal-like structure. Further we found that expression of GmSEP1 was altered during the course of seed development and more accumulated in outer and inner integuments, epidermis and endothelium at the globular stage, heart stage and cotyledon stage and in seed coat parenchyma at the early maturation stage of seed development. Taken together, GmSEP1 might play important role in soybean reproductive development, in particular, in petal and seed coat development. PMID- 19289161 TI - The complete mitochondrial genome of Watersipora subtorquata (Bryozoa, Gymnolaemata, Ctenostomata) with phylogenetic consideration of Bryozoa. AB - The phylogenetic position of the Bryozoa has long been controversial. In this paper, we have determined the complete mitochondrial genome of the Watersipora subtorquata (Bryozoa, Gymnolaemata, Ctenostomata). It is a circular molecule of 14,144 bp, relatively small compared with most other metazoan mitochondrial genomes, and bears some unusual features. All genes in the W. subtorquata mtDNA, unlike those in two bryozoan mtDNAs and most other metazoan mtDNAs published previously, are transcribed from the same strand. It has a unique gene order which differs radically from that of other metazoans. Drastic gene rearrangements were also found among bryozoan mtDNAs. To investigate the phylogenetic position of Bryozoa, analyses based on amino acid sequences of 11 protein-coding genes (excluding atp6 and atp8) from 25 metazoan mtDNAs were made utilizing ML and Bayesian methods. Lophotrochozoa was recovered as monophyletic with strong support in our analyses. Lophophorate was undoubted within Lophotrochozoa, but appears as polyphyletic, which indicates that the lophophores of this group may be of different origin. The existence of Phoronozoa was rejected. Our analyses indicated that Phoronida is more closely related to Annelid instead of Brachiopod. Chaetognatha appeared as the sister group of Bryozoa and they formed a clade together with strong support. More evidence is needed to clarify the relationship of these two phyla. PMID- 19289162 TI - Protective effect of supercritical fluid rosemary extract, Rosmarinus officinalis, on antioxidants of major organs of aged rats. AB - Rosemary leaves, "Rosmarinus officinalis", possess a variety of antioxidant, anti tumoral and anti-inflammatory bioactivities. We hypothesized that rosemary extract could enhance antioxidant defenses and improve antioxidant status in aged rats. This work evaluates whether supplementing their diet with supercritical fluid (SFE) rosemary extract containing 20% antioxidant carnosic acid (CA) reduces oxidative stress in aged rats. Aged Wistar rats (20 months old) were included in the study. Rats were fed for 12 weeks with a standard kibble (80%) supplemented with turkey breast (20%) containing none or one of two different SFE rosemary concentrations (0.2% and 0.02%). After sacrifice, tissue samples were collected from heart and brain (cortex and hippocampus). Enzyme activities of catalase (CAT), glutathione peroxidase (GPX), superoxide dismutase (SOD) and nitric oxide synthase (NOS) were quantitatively analyzed. Lipid peroxidation and levels of reactive oxygen species (ROS) were also determined. Rosemary decreased lipid peroxidation in both brain tissues. The levels of catalase activities in heart and cortex were decreased in the rosemary-treated groups. The SFE rosemary treated rats presented lower NOS levels in heart and lower ROS levels in hippocampus than the control rats. Supplementing the diet of aged rats with SFE rosemary extract produced a decrease in antioxidant enzyme activity, lipid peroxidation and ROS levels that was significant for catalase activity in heart and brain, NOS in heart, and LPO and ROS levels in different brain tissues. These observations suggest that the rosemary supplement improved the oxidative stress status in old rats. PMID- 19289163 TI - Anaerobic digestion of microalgae as a necessary step to make microalgal biodiesel sustainable. AB - The potential of microalgae as a source of biofuels and as a technological solution for CO2 fixation is subject to intense academic and industrial research. In the perspective of setting up massive cultures, the management of large quantities of residual biomass and the high amounts of fertilizers must be considered. Anaerobic digestion is a key process that can solve this waste issue as well as the economical and energetic balance of such a promising technology. Indeed, the conversion of algal biomass after lipid extraction into methane is a process that can recover more energy than the energy from the cell lipids. Three main bottlenecks are identified to digest microalgae. First, the biodegradability of microalgae can be low depending on both the biochemical composition and the nature of the cell wall. Then, the high cellular protein content results in ammonia release which can lead to potential toxicity. Finally, the presence of sodium for marine species can also affect the digester performance. Physico chemical pretreatment, co-digestion, or control of gross composition are strategies that can significantly and efficiently increase the conversion yield of the algal organic matter into methane. When the cell lipid content does not exceed 40%, anaerobic digestion of the whole biomass appears to be the optimal strategy on an energy balance basis, for the energetic recovery of cell biomass. Lastly, the ability of these CO2 consuming microalgae to purify biogas and concentrate methane is discussed. PMID- 19289164 TI - RNA interference technology to improve recombinant protein production in Chinese hamster ovary cells. AB - RNA interference (RNAi) technology has become a novel tool for silencing gene expression in cells or organisms, and has also been used to develop new therapeutics for certain diseases. This review describes its other application of using RNAi technology to increase cellular productivity and the quality of recombinant proteins that are produced in Chinese hamster ovary (CHO) cells, the most important mammalian cell line used in producing licensed biopharmaceuticals in these days. The approaches reported include the silencing of apoptosis associated gene expression, protein glycosylation-associated gene expression, lactate dehydrogenase involved in cellular metabolism, and dihydrofolate reductase used for gene amplification. All of these works belong to the single component approach therefore depends strongly on the identification of the down regulation of the critical target gene which can markedly influence the cellular functions associated with recombinant protein expression in CHO cells. Future RNAi approaches can be extended to silence multiple targets involved in different cellular pathways for changing the global gene regulation in cells, as well as the targets related to microRNA molecules for cellular self regulation. PMID- 19289165 TI - Response to chronic exposure to hexavalent chromium in human monocytes. AB - Elevated circulating levels of metal ions, particularly chromium, have been measured in the blood of patients with metal hip implants, and this has lead to concerns about the long term safety of the prostheses. For example, depletion of lymphocytes has been reported in vivo in patients with metallic prostheses, and correlated with elevated chromium and cobalt concentrations in blood. However, the implications for immune function are unclear. We have assessed the in vitro responses of U937 human monocytes to chronic exposure (4 weeks) to Cr (VI) ions at concentrations which have been measured in patients with metal artificial hip implants (0.05-0.5 microM). Chronic exposure to these low clinically relevant concentrations of Cr (VI) induced a potent adaptive response with elevated glutathione-S-transferase (pi) expression and increased activities and expression of reactive oxygen scavengers, superoxide dismutases, catalase and glutathione peroxidase. Such direct toxicity of Cr ions may contribute to the effects of metal implants on lymphocyte populations in vivo. PMID- 19289166 TI - Construction and characterization of subtractive stage-specific expressed sequence tag (EST) libraries of the pinewood nematode Bursaphelenchus xylophilus. AB - To establish expressed sequence tag databases of the two life stages (the dispersal and propagative stages) of pinewood nematode Bursaphelenchus xylophilus, subtractive EST libraries that were specific to the dispersal 4th larval stage (D4S) and the pine-grown propagative mixed (PGPS) stage were constructed by suppressed subtractive hybridization, and annotated by BLASTx and Gene Ontology (GO). A total of 1112 (57.7%) contigs from the D4S-cDNA library and 1215 (46.7%) contigs from the PGPS-specific cDNA libraries had matched BLASTx hits (E], using morphological and cytological approaches. Symphyotrichum is the largest and most complex genus within the subtribe and includes four subgenera: Symphyotrichum (x=7, 8), Virgulus (x=4, 5), Astropolium (x=5), and Chapmaniani (x=7). In this study we used two nuclear markers, the nrDNA internal transcribed spacer (ITS) and the low-copy nuclear gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH), to resolve intergeneric and interspecific relationships within the subtribe at the diploid level, and to determine whether our phylogenies validate the classifications of Nesom or Semple. Our results confirm the distinct generic status of Canadanthus and Ampelaster, whereas Psilactis and Almutaster form a polytomy with Symphyotrichum. Within Symphyotrichum, subg. Virgulus is monophyletic based on ITS and appears polyphyletic based on GAPDH. Neither the ITS nor the GAPDH analyses support a distinct status for subg. Astropolium, which groups within subg. Symphyotrichum. In general, interspecific relationships within Symphyotrichum are unresolved. Lack of resolution may be interpreted as a case of recent and rapid evolutionary radiation. PMID- 19289176 TI - Overexpression of E2F-1 inhibits progression of gastric cancer in vitro. AB - E2F-1 plays a critical role in cell cycle regulation and other biological processes in cells. E2F-1 mediates apoptosis and suppresses tumorigenesis in many tissue types, but there are few data available on E2F-1 expression and its relationship to tumor kinetics in gastric cancer. To gain better insight into the involvement of E2F-1 in the biological characteristics of gastric tumors, we investigated the effect of E2F-1 overexpression on the progression of gastric carcinoma cells. A gastric cancer cell line stably overexpressing E2F-1 (MGC 803/E2F-1) was established. The influence of E2F-1 overexpression on in vitro cell growth was assessed by measuring cell survival, colony formation, and cell cycle progression. The results clearly show that overexpression of E2F-1 significantly inhibits cell growth and proliferation, blocking entry into the S phase of the cell cycle. MGC-803/E2F-1 cells also had a higher apoptotic rate than control cells. In addition, E2F-1 reduced the motility and invasion of gastric cancer cells. PMID- 19289177 TI - Characterization and sequence analysis of manganese superoxide dismutases from Brachyura (Crustacea: Decapoda): hydrothermal Bythograeidae versus littoral crabs. AB - Hydrothermal vent conditions are particular and organisms living in these environments may have developed detoxification mechanisms and/or genetic adaptations. In particular, physico-chemical conditions are thought to generate reactive oxygen species, highly toxic for organisms. The enzyme superoxide dismutase constitutes the first line of defense against oxidative damage. To improve our understanding of the environmental impacts exerted on the vent organisms, we have characterized the two manganese superoxide dismutase cDNAs (mitochondrial: mMnSOD and cytoplasmic: cMnSOD) of three members of the Bythograeidae (Bythograea thermydron, Cyanagraea praedator and Segonzacia mesatlantica), the only endemic crab family living in hydrothermal vents. In comparison, the isolation of manganese superoxide dismutase cDNAs was also carried out in several littoral crab families. MnSOD signatures were found in both sequences from each species studied, as well as different residues involved in metal coordination and protein activity. The phylogenetic analysis performed confirms the probable ancient duplication that gave rise to the two MnSODs (cMnSOD and mMnSOD). This study describes two potential distinct mMnSOD isoforms presenting particular peptide signals. Nevertheless, no sequence particularity that could support the hypothesis of a genetic adaptation was found in Bythograeidae's MnSODs compared to the other sequences. The mRNA expression analysis performed by real-time PCR on B. thermydron and S. mesatlantica compared to Cancer pagurus and Necora puber revealed a higher cMnSOD and mMnSOD mRNA expression in hydrothermal crabs compared to littoral crabs. PMID- 19289178 TI - Patients with chronic hepatitis B infection display deficiency of plasmacytoid dendritic cells with reduced expression of TLR9. AB - Chronic hepatitis B virus (HBV) infection is a complex interaction between replicating noncytopathic virus and dysregulatory host antiviral immunity. Plasmacytoid dendritic cells (pDCs) contribute to innate antiviral immunity via secreting type I interferons. Toll-like receptor (TLR) 9 is involved in major pattern recognition receptors expressed in pDCs. The frequency of pDCs and TLR9 expression in peripheral blood mononuclear cells (PBMC) was determined, using flow cytometry. IFN-alpha production by PBMC was evaluated in vitro in the presence of cytidine phosphate guanosine (CpG) with/without pDCs. The correlation between TLR9, pDCs frequency and viral load was also evaluated. TLR9 expression in pDCs in chronic HBV patients was significantly ( approximately 50%) reduced, supported by approximately 70% reduction of TLR9 mRNA, in comparison to healthy controls, correlating with the impairment of IFN-alpha production in vitro. Furthermore, pDCs frequency in these patients was substantially reduced ( approximately 30%), inversely correlating with serum ALT levels and HBV viral load. HBsAg and HBcAg were detected by immunohistochemistry in pDCs in chronic HBV patients. We conclude that HBV infection results in reduced frequency of circulating pDCs and their functional impairment via inhibiting the expression of TLR9. These data may provide useful information in both basic research and clinical treatment of chronic HBV infection. PMID- 19289179 TI - Phospholipase D- and phosphatidic acid-mediated signaling in plants. AB - The phospholipase D (PLD) family in higher plants is composed of multiple members, and each of the Arabidopsis PLDs characterized displays distinguishable properties in activity regulation and/or lipid preferences. The molecular and biochemical heterogeneities of the plant PLDs play important roles in the timing, location, and amount of phosphatidic acid (PA) produced. PLD-catalyzed production of PA has been shown to play important roles in plant growth, development, and response to various stresses, including drought, salinity, freezing, and nutrient deficiency. PLD and PA affect cellular processes through different modes of action, including direct target protein binding and biophysical effects on cell membranes. Improved knowledge on the mechanism by which specific PLDs and PA mediate given plant responses will facilitate the understanding of the molecular processes that connect the stimulus perception on membranes to intracellular actions and physiological responses. PMID- 19289180 TI - Phospholipase D in calcium-regulated exocytosis: lessons from chromaffin cells. AB - Membrane fusion remains one of the less well-understood processes in cell biology. A variety of mechanisms have been proposed to explain how the generation of fusogenic lipids at sites of exocytosis facilitates secretion in mammalian cells. Over the last decade, chromaffin cells have served as an important cellular model to demonstrate a key role for phospholipase D1 (PLD1) generated phosphatidic acid in regulated exocytosis. The current model proposes that phosphatidic acid plays a biophysical role, generating a negative curvature and thus promoting fusion of secretory vesicles with the plasma membrane. Moreover, multiple signaling pathways converging on PLD1 regulation have been unraveled in chromaffin cells, suggesting a complex level of regulation dependant on the physiological context. PMID- 19289181 TI - Interferon-beta modifies the peripheral blood cell cytokine secretion in patients with multiple sclerosis. AB - Immunotherapy with Interferon-beta (IFNbeta) results in remarkably beneficial effects in patients with relapsing-remitting multiple sclerosis (MS), although the mechanisms by which it exerts these beneficial effects remain poorly understood. An investigation was made of the effects of IFNbeta on pro inflammatory and anti-inflammatory cytokine production in peripheral blood cells in MS patients, both untreated and those undergoing immunotherapy, as well as in healthy controls. Results show a significant increase in the production of pro inflammatory cytokines such as TNFalpha, IFNgamma and IL-12 in the plasma and in the supernatant of leukocyte cultures from MS patients with the untreated disease; IFNbeta administration significantly reduced the levels of TNFalpha and IFNgamma, with no changes in the level of IL-12. The Interferon-beta therapy also led to a significant increase in the production of IL-10, as well as a slight increase in that of TGFbeta. The reduction in pro-inflammatory cytokine production in the treated MS patient group, accompanied by a simultaneous increase in the production of anti-inflammatory cytokines and the reduction of relapse rates suggests that the beneficial effects of IFNbeta immunotherapy result, at least in part, from the modulation of cytokine patterns. PMID- 19289182 TI - Assembly and transport mechanism of tripartite drug efflux systems. AB - Multidrug efflux (MDR) pumps remove a variety of compounds from the cell into the external environment. There are five different classes of MDR pumps in bacteria, and quite often a single bacterial species expresses multiple classes of pumps. Although under normal circumstances MDR pumps confer low-level intrinsic resistance to drugs, the presence of drugs and mutations in regulatory genes lead to high level expression of MDR pumps that can pose problems with therapeutic treatments. This review focuses on the resistance nodulation cell division (RND) class of MDR pumps that assemble from three proteins. Significant recent advancement in structural aspects of the three pump components has shed new light on the mechanism by which the tripartite efflux pumps extrude drugs. This new information will be critical in developing inhibitors against MDR pumps to improve the potency of prescribed drugs. PMID- 19289183 TI - Recent developments in liver pathology. AB - Progress in liver histopathology continues to be made, as evidenced by recent publications in all areas of hepatobiliary disease. Multinucleated giant hepatocytes, known to be associated with autoimmune and drug hepatitis, now have been seen in chronic hepatitis C with or without HIV infection and in patients with infection by human herpesvirus-6A. The new term Mallory-Denk body (formerly the Mallory body) recognizes the substantial contributions to this field by Professor Helmut Denk of Austria. The problems of fatty liver and hepatic iron overload have been addressed in studies highlighting their complex pathogenesis. Genomic and immunohistochemical analysis of liver tumors provides important diagnostic information, particularly regarding the use of glypican-3 in the diagnosis of hepatocellular carcinoma. PMID- 19289184 TI - Recommendations for the reporting of surgically resected specimens of renal cell carcinoma: the Association of Directors of Anatomic and Surgical Pathology. AB - A checklist based approach to reporting the relevant pathologic details of renal cell carcinoma resection specimens improves the completeness of the report. Karyotypic evaluation of renal neoplasms has refined but also complicated their classification. The number of diagnostic possibilities has increased and the importance of distinguishing different tumor types has been underscored by dramatic variation in prognosis and the development of targeted therapies for specific subtypes. The increasing number of recognized renal neoplasms has implications for handling renal resection specimens. Furthermore, the prognostic significance of other features of renal neoplasms related to grade and stage has been demonstrated. This guideline for the handling of renal resection specimens will focus on problem areas in the evolving practice of diagnosis, grading, and staging of renal neoplasms. The accompanying checklist will serve to ensure that all necessary details of the renal resection specimen are included in the surgical pathology report. PMID- 19289185 TI - Pathologic findings in 200 consecutive fallopian tube specimens. PMID- 19289186 TI - Expression of caspase-3 in enteric cells is related to development of chagasic megacolon. PMID- 19289187 TI - Natural history of host-parasite interactions. Preface. PMID- 19289188 TI - HLA-mediated control of HIV and HIV adaptation to HLA. AB - The human immunodeficiency virus (HIV) epidemic provides a rare opportunity to examine in detail the initial stages of a host-pathogen co-evolutionary struggle in humans. The genes encoding the human leukocyte antigen (HLA) class I molecules have a critical influence in the success or failure of the immune response against HIV. The particular HLA class I molecules expressed by each individual defines the type of cytotoxic T-lymphocyte (CTL) response that is made against the virus. This chapter describes the role of HLA class I and the CTL response in controlling HIV replication, and discusses the extent to which HIV has already adapted to those HLA class I molecules and CTL responses that are most effective in viral suppression. It is evident that viral mutations that enable HIV to evade the CTL response are indeed already accumulating in populations where the selecting HLA molecules are highly prevalent, indicating the dynamic and shifting nature of the evolutionary interplay between HIV and human populations. PMID- 19289189 TI - An evolutionary perspective on parasitism as a cause of cancer. AB - For the past half-century, the dominant paradigm of oncogenesis has been mutational changes that disregulate cellular control of proliferation. Parasitic causes of cancer were first incorporated into this paradigm by suggesting mechanisms through which parasitism might increase mutational damage, such as generation of mutagenic compounds during immunological activity. The growing recognition of the molecular mechanisms of pathogen-induced oncogenesis and the difficulty of generating oncogenic mutations without first having large populations of dysregulated cells, however, suggests that pathogens, particularly viruses, are major initiators of oncogenesis for many if not most cancers, and that the traditional mutation-driven process becomes the dominant process after this initiation. Molecular phylogenies of individual cancers should facilitate testing of this idea and the identification of causal pathogens. PMID- 19289190 TI - Invasion of the body snatchers: the diversity and evolution of manipulative strategies in host-parasite interactions. AB - Parasite-induced alteration of host behaviour is a widespread transmission strategy among pathogens. Understanding how it works is an exciting challenge from both a mechanistic and an evolutionary perspective. In this review, we use key examples to examine the proximate mechanisms by which parasites are known to control the behaviour of their hosts. Special attention is given to the recent developments of post-genomic tools, such as proteomics, for determining the genetic basis of parasitic manipulation. We then discuss two novel perspectives on host manipulation (mafia-like strategy and exploitation of host compensatory responses), arguing that parasite-manipulated behaviours could be the result of compromises between host and parasite strategies. Such compromises may occur when collaborating with the parasite is less costly for the host in terms of fitness than is resisting parasite-induced changes. Therefore, even when changes in host behaviour benefit the parasite, the host may still play some role in the switch in host behaviour. In other words, the host does not always become part of the parasite's extended phenotype. For example, parasites that alter host behaviour appear to induce widely disseminated changes in the hosts' central nervous system, as opposed to targeted attacks on specific neural circuits. In some host parasite systems, the change in host behaviour appears to require the active participation of the host (e.g., via host immune-neural connections). Even when the change in host behaviour results in clear fitness benefits for the parasite, these behavioural changes may sometimes be produced by the host. Changes in host behaviour that decrease the fitness costs of infection could be selected for, even if these changes also benefit the parasite. PMID- 19289191 TI - Evolutionary drivers of parasite-induced changes in insect life-history traits from theory to underlying mechanisms. AB - Many hosts are able to tolerate infection by altering life-history traits that are traded-off one against another. Here the reproductive fitness of insect hosts and vectors is reviewed in the context of theories concerning evolutionary mechanisms driving such alterations. These include the concepts that changes in host reproductive fitness are by-products of infection, parasite manipulations, host adaptations, mafia-like strategies or host compensatory responses. Two models are examined in depth, a tapeworm/beetle association, Hymenolepis diminuta/Tenebrio molitor and malaria infections in anopheline mosquitoes. Parasite-induced impairment of vitellogenesis ultimately leads to a decrease in female reproductive success in both cases, though by different means. Evidence is put forwards for both a manipulator molecule of parasite origin and for host initiated regulation. These models are backed by other examples in which mechanisms underlying fecundity reduction or fecundity compensation are explored. It is concluded that evolutionary theories must be supported by empirical evidence gained from studying molecular, biochemical and physiological mechanisms underlying changes in host life-history traits, ideally using organisms that have evolved together and that are in their natural environment. PMID- 19289192 TI - Ecological immunology of a tapeworms' interaction with its two consecutive hosts. AB - Host-parasite interactions in parasites with complex life cycles have recently gained much interest. Here, we take an evolutionary ecologist's perspective and analyse the immunological interaction of such a parasite, the model tapeworm Schistocephalus solidus, with its two intermediate hosts, a cyclopoid copepod and the three-spined stickleback. We will be focussing especially on the parallel links between the different phases during an infection in the different hosts; the immunological interactions between host(s) and parasite; and their impact on parasite establishment, growth, host manipulation and parasite virulence in the next host in the cycle. We propose to extend the 'extended phenotype' concept and not only include the ultimate but also the proximate, physiological causes. In particular, parasite-induced host manipulation is suggested to be caused by the interactions of the parasite with the hosts' immune systems. PMID- 19289193 TI - Tracking transmission of the zoonosis Toxoplasma gondii. AB - Toxoplasma gondii is a highly successful parasite that infects many host species and has colonised a wide range of habitats. Review of the parasite's life cycle demonstrates that it has become adapted to exploit multiple routes of transmission through a sexual cycle in the definitive host and asexually, through carnivory, and by vertical transmission. These alternative routes may operate synergistically to enhance transmission, but they might also provide a vehicle for selection leading to partitioning of strains in the environment. Genetic analysis has shown that parasite population structure varies globally. In South America, there is high strain diversity while in North America, Europe and Africa three clonal strain types predominate. This may imply a shift from sexual to asexual transmission. Mapping of the parasite genome has provided a wealth of markers for strain characterisation. Close genotyping of isolates gives evidence of multiple infection and recombination in natural populations and reveals differences in both the distribution and the phenotype of strains. More intensive epidemiological studies are now required to unravel the networks of transmission operating within defined habitats. PMID- 19289194 TI - Parasites and biological invasions. AB - There is considerable current interest in the role that parasites can play in biological invasions. This review looks at the fate of parasites during a biological invasion and at their impact on native and invasive hosts, and asks whether parasites can mediate invasion success. An introduced species may lose its parasites as a result of the introduction and such release from its natural enemies may be an important factor determining invasion success. In addition, an introduced species may acquire parasites from its new environment or it may introduce novel parasites to hosts in the new range. As a result of local adaptation, parasites tend to have a differential effect on native versus invading hosts. The relative impact on the fitness of natives and invaders can be important for the outcome of an invasion and may, for example, reverse the pattern of competitive dominance seen in uninfected hosts. Parasites may mediate invasion success through their effect on host fitness and thus on host population growth and stability. Furthermore, by modifying host-host interactions (including competition and predation), parasites can be important factors that determine the success of an invasion and its impact on the recipient community. PMID- 19289195 TI - Zoonoses in wildlife integrating ecology into management. AB - Zoonoses in wildlife not only play an important ecological role, but pose significant threats to the health of humans, domestic animals and some endangered species. More than two-thirds of emerging, or re-emerging, infectious diseases are thought to originate in wildlife. Despite this, co-ordinated surveillance schemes are rare, and most efforts at disease control operate at the level of crisis management. This review examines the pathways linking zoonoses in wildlife with infection in other hosts, using examples from a range of key zoonoses, including European bat lyssaviruses and bovine tuberculosis. Ecologically based control, including the management of conditions leading to spill-overs into target host populations, is likely to be more effective and sustainable than simple reductions in wildlife populations alone. PMID- 19289196 TI - Understanding the interaction between an obligate hyperparasitic bacterium, Pasteuria penetrans and its obligate plant-parasitic nematode host, Meloidogyne spp. AB - Pasteuria penetrans is an endospore-forming bacterium, which is a hyperparasite of root-knot nematodes Meloidogyne spp. that are economically important pests of a wide range of crops. The life cycle of the bacterium and nematode are described with emphasis on the bacterium's potential as a biocontrol agent. Two aspects that currently prohibit the commercial development of the bacterium as a biocontrol agent are the inability to culture it outside its host and its host specificity. Vegetative growth of the bacterium is possible in vitro; however, getting the vegetative stages of the bacterium to enter sporogenesis has been problematic. Insights from genomic survey sequences regarding the role of cation concentration and the phosphorylation of Spo0F have proved useful in inducing vegetative bacteria to sporulate. Similarly, genomic data have also proved useful in understanding the attachment of endospores to the cuticle of infective nematode juveniles, and a Velcro-like model of spore attachment is proposed that involves collagen-like fibres on the surface of the endospore interacting with mucins on the nematode cuticle. Ecological studies of the interactions between Daphnia and Pasteuria ramosa are examined and similarities are drawn between the co-evolution of virulence in the Daphnia system and that of plant-parasitic nematodes. PMID- 19289197 TI - Host-parasite relations and implications for control. AB - This paper considers the various measures available to control several of the neglected tropical diseases (NTDs). To develop the optimum methods for controlling the parasites that cause these NTDs, knowledge of the life cycles of both the parasites and their vectors are essential. Each NTD requires its own strategy for control based on detailed knowledge of the life cycle, and vector control, chemotherapy, better water supplies and better hygiene are all components that may be appropriate. For some diseases, improved drugs are urgently required, for some the tools are available for elimination, while uniquely guinea worm could be eradicated without any chemotherapeutic drug being used. Several NTDs lend themselves to mass drug administration (MDA) in which human populations are annually offered safe, effective and usually donated drugs with a view to morbidity control and/or elimination. The drugs could and should be used to improve the quality of millions of lives, prevent suffering, stigma, disfigurement and early death. The role of pharmaceutical companies who have donated their drugs for the treatment of millions of disadvantaged people in the developing world is acknowledged. One result of such drug pressure however is that evolutionary change may result, and it is incumbent on scientists during monitoring and evaluation of control programmes to ensure that such changes are recognised. One other unfortunate development is that a paucity of newly trained vector-borne disease experts may constrain future control efforts. PMID- 19289198 TI - Onchocerca-Simulium interactions and the population and evolutionary biology of Onchocerca volvulus. AB - Parasite-vector interactions shape the population dynamics of vector-borne infections and contribute to observed epidemiological patterns. Also, parasites and their vectors may co-evolve, giving rise to locally adapted combinations or complexes with the potential to stabilise the infection. Here, we focus on Onchocerca-Simulium interactions with particular reference to the transmission dynamics of human onchocerciasis. A wide range of simuliid species may act as vectors of Onchocerca volvulus, each exerting their own influence over the local epidemiology and the feasibility of controlling/eliminating the infection. Firstly, current understanding of the processes involved in parasite acquisition by, and development within, different Simulium species in West Africa and Latin America will be reviewed. A description of how Onchocerca and Simulium exert reciprocal effects on each other's survival at various stages of the parasite's life cycle within the blackfly, and may have adapted to minimise deleterious effects on fitness and maximise transmission will be given. Second, we describe the interactions in terms of resultant (positive and negative) density-dependent processes that regulate parasite abundance, and discuss their incorporation into mathematical models that provide useful qualitative insight regarding transmission breakpoints. Finally, we examine the interactions' influence upon the evolution of anthelmintic resistance, and conclude that local adaptation of Onchocerca-Simulium complexes will influence the feasibility of eliminating the parasite reservoir in different foci. PMID- 19289200 TI - Biothermodynamics, Part A. Preface. PMID- 19289199 TI - Microsporidians as evolution-proof agents of malaria control? AB - Despite our efforts at malaria control, malaria remains one of our most serious and deadly diseases. The failure of control stems in part from the parasite's intense transmission in many areas and from the emergence and spread of resistance of the malaria parasites and their mosquito vectors against most of the chemicals used to attack them. New methods for control are desperately needed. However, new methods will be useful only if they are effective (i.e., decrease transmission substantially) and evolutionarily sustainable (i.e., evolution-proof, in that they prevent evolution from eroding efficacy). We suggest microsporidian parasites that infect mosquitoes could be potentially effective and sustainable agents for malaria control. They may be effective because they target several epidemiologically important traits: survival of larvae (and thus number of adult mosquitoes), adult longevity, biting rate and the development of malaria within the mosquitoes. Even if each trait is affected only moderately, the intensity of transmission can be reduced considerably. They may be evolution-proof, for the evolutionarily most important trait is juvenile survival, whereas the two epidemiologically most important factors are traits of the adult mosquito: biting rate and longevity. Under the intense microsporidian pressure of a control programme, it is likely (if not inevitable) that the larvae evolve to survive microsporidian infection. However, if this larval tolerance to microsporidians is genetically correlated with the adult traits, tolerant mosquitoes may not live as long and bite less frequently than microsporidian sensitive ones. While such a trade-off has not been measured, combining several studies suggests indirectly a negative genetic correlation between larval tolerance and adult longevity. Therefore, evolution might not undermine control; rather it might increase its effectiveness. While the evolution of resistance may be inevitable, the failure of control need not be. PMID- 19289201 TI - Practical approaches to protein folding and assembly: spectroscopic strategies in thermodynamics and kinetics. AB - We describe here the use of several spectroscopies, such as fluorescence emission, circular dichroism, and differential quenching by acrylamide, in examining the equilibrium and kinetic folding of proteins. The first section regarding equilibrium techniques provides practical information for determining the conformational stability of a protein. In addition, several equilibrium folding models are discussed, from two-state monomer to four-state homodimer, providing a comprehensive protocol for interpretation of folding curves. The second section focuses on the experimental design and interpretation of kinetic data, such as burst-phase analysis and exponential fits, used in elucidating kinetic folding pathways. In addition, simulation programs are used routinely to support folding models generated by kinetic experiments, and the fundamentals of simulations are covered. PMID- 19289202 TI - Using thermodynamics to understand progesterone receptor function: method and theory. AB - Progesterone receptors (PRs) are members of the nuclear receptor superfamily of ligand-activated transcription factors. The mechanisms by which receptors such as PR assemble at a promoter and recruit coactivators are well understood at the biochemical level. However, a rigorous and thus quantitatively predictive understanding of function is entirely lacking. This is so in part because the study of receptor function has largely been carried out using semiquantitative or qualitative approaches. These types of analyses are limited in their ability to resolve thermodynamically valid and physically meaningful microscopic interaction parameters. This includes resolution of intrinsic binding constants and cooperativity terms, as well as the mathematical framework for integrating these values into a larger molecular code for function. Here we present our experimental and theoretical approach for dissecting the linked reactions associated with PR and coactivator assembly at complex promoter sequences. We discuss the use of analytical ultracentrifugation and quantitative DNase footprint titration and their coupling to exact theoretical treatments. We then highlight the major findings of these studies and their implications for understanding and reevaluating receptor function. PMID- 19289203 TI - Direct quantitation of Mg2+-RNA interactions by use of a fluorescent dye. AB - The ionic composition of a solution strongly influences the folding of an RNA into its native structure; of particular importance, the stabilities of RNA tertiary structures are sharply dependent on the concentration of Mg2+. Most measurements of the extent of Mg2+ interaction with an RNA have relied on equilibrium dialysis or indirect measurements. Here we describe an approach, based on titrations in the presence of a fluorescent indicator dye, that accurately measures the excess Mg2+ ion neutralizing the charge of an RNA (the interaction or Donnan coefficient, Gamma2+) and the total free energy of Mg2+-RNA interactions (DeltaG(RNA-2+)). Automated data collection with computer-controlled titrators enables the collection of much larger data sets in a short time, compared to equilibrium dialysis. Gamma2+ and DeltaG(RNA-2+) are thermodynamically rigorous quantities that are directly comparable with the results of theoretical calculations and simulations. In the event that RNA folding is coupled to the addition of MgCl2, the method directly monitors the uptake of Mg2+ associated with the folding transition. PMID- 19289204 TI - Analysis of repeat-protein folding using nearest-neighbor statistical mechanical models. AB - The linear "Ising" model, which has been around for nearly a century, treats the behavior of linear arrays of repetitive, interacting subunits. Linear "repeat proteins" have only been described in the last decade or so, and their folding energies have only been characterized very recently. Owing to their repetitive structures, linear repeat-proteins are particularly well suited for analysis by the nearest-neighbor Ising formalism. After briefly describing the historical origins and applications of the Ising model to biopolymers, and introducing repeat protein structure, this chapter will focus on the application of the linear Ising model to repeat proteins. When applied to homopolymers, the model can be represented and applied in a fairly simplified form. When applied to heteropolymers, where differences in energies among individual subunits (i.e. repeats) must be included, some (but not all) of this simplicity is lost. Derivations of the linear Ising model for both homopolymer and heteropolymer repeat-proteins will be presented. With the increased complexity required for analysis of heteropolymeric repeat proteins, the ability to resolve different energy terms from experimental data can be compromised. Thus, a simple matrix approach will be developed to help inform on the degree to which different thermodynamic parameters can be extracted from a particular set of unfolding curves. Finally, we will describe the application of these models to analyze repeat-protein folding equilibria, focusing on simplified repeat proteins based on "consensus" sequence information. PMID- 19289205 TI - Isothermal titration calorimetry: general formalism using binding polynomials. AB - The theory of the binding polynomial constitutes a very powerful formalism by which many experimental biological systems involving ligand binding can be analyzed under a unified framework. The analysis of isothermal titration calorimetry (ITC) data for systems possessing more than one binding site has been cumbersome because it required the user to develop a binding model to fit the data. Furthermore, in many instances, different binding models give rise to identical binding isotherms, making it impossible to discriminate binding mechanisms using binding data alone. One of the main advantages of the binding polynomials is that experimental data can be analyzed by employing a general model-free methodology that provides essential information about the system behavior (e.g., whether there exists binding cooperativity, whether the cooperativity is positive or negative, and the magnitude of the cooperative energy). Data analysis utilizing binding polynomials yields a set of binding association constants and enthalpy values that conserve their validity after the correct model has been determined. In fact, once the correct model is validated, the binding polynomial parameters can be immediately translated into the model specific constants. In this chapter, we describe the general binding polynomial formalism and provide specific theoretical and experimental examples of its application to isothermal titration calorimetry. PMID- 19289206 TI - Kinetic and equilibrium analysis of the myosin ATPase. AB - The myosin superfamily consists of more than 35 classes (each consisting of multiple isoforms) that have diverse cellular activities. The reaction pathway of the actin-activated myosin ATPase appears to be conserved for all myosin isoforms, but the rate and equilibrium constants that define the ATPase pathway vary significantly across the myosin superfamily, resulting in kinetic differences that that allow myosins to carry out diverse mechanical functions. Therefore, it is important to determine the lifetimes and relative populations of the key biochemical intermediates to obtain an understanding of a particular myosin's cellular function. This chapter provides procedures for determining the overall and individual rate and equilibrium constants of the actomyosin ATPase cycle, including actomyosin binding and dissociation, ATP binding, ATP hydrolysis, phosphate release, and ADP release and binding. Many of the methods described in the chapter are applicable to the characterization of other ATPase enzymes. PMID- 19289207 TI - The Hill coefficient: inadequate resolution of cooperativity in human hemoglobin. AB - The Hill coefficient nH is a dimensionless parameter that has long been used as a measure of the extent of cooperativity. Originally derived from the oxygen binding curve of human hemoglobin (Hb) by A. V. Hill in 1910, and reinvented by J. Wyman several decades later, nH is indexed to the stoichiometry of ligation and is indirectly related to the overall cooperative free energy for binding all four oxygen ligands. However, the overall cooperative free energy of Hb ligation can be measured directly by experimental methods. The microscopic cooperative free energies that relate to energetic coupling between specific subunit pairs can also be experimentally determined, while the Hill coefficient is, by its nature, a macroscopic parameter that cannot detect differences among specific subunit-subunit couplings. Its continued use in studies of the mechanism of cooperativity in Hb is therefore of increasingly limited value. PMID- 19289208 TI - Methods for measuring the thermodynamic stability of membrane proteins. AB - Learning how amino acid sequences define protein structure has been a major challenge for molecular biology since the first protein structures were determined in the 1960s. In contrast to the staggering progress with soluble proteins, investigations of membrane protein folding have long been hampered by the lack of high-resolution structures and the technical challenges associated with studying the folding process in vitro. In the past decade, however, there has been an explosion of new membrane protein structures and a slower but notable increase in efforts to study the factors that define these structures. Here we review the methods that have been used to evaluate the thermodynamic stability of membrane proteins and provide some salient examples of how the methods have been used to begin to understand the energetics of membrane protein folding. PMID- 19289209 TI - NMR analysis of dynein light chain dimerization and interactions with diverse ligands. AB - NMR is a powerful tool for quantitative measurement of the thermodynamic properties of biological systems. In this review, we discuss the role NMR has played in understanding the various coupled equilibria in dimerization of dynein light chain LC8 and in its interactions with its ligands. LC8, a very highly conserved 89-residue homodimer also known as DYNLL, is an essential component of the dynein and Myosin V molecular motors and is also found in various other complexes. LC8 binds to disordered segments of its partners, promoting them to dimerize and form more ordered structures, often coiled coils. The monomer-dimer equilibrium is controlled by electrostatic interactions at the dimer interface, such as by phosphorylation of residue Ser88, which is a regulatory mechanism for LC8 in vivo. NMR experiments have uncovered several subtle interactions--weak dimerization of a phosphomimetic mutant, and allosteric interaction between the LC8 binding sites--that have been overlooked by other methods. NMR has also provided a residue-specific view of the titration of histidine residues at the LC8 dimer interface, and of a nascent helix in one of the binding partners, the primarily disordered dynein intermediate chain IC74. We give special attention to methods for quantitative interpretation of NMR spectra, an important consideration when using NMR to measure equilibria. PMID- 19289210 TI - Characterization of parvalbumin and polcalcin divalent ion binding by isothermal titration calorimetry. AB - The elucidation of structure-affinity relationships in EF-hand proteins requires a reliable assay of divalent ion affinity. In principle, isothermal titration calorimetry (ITC) should be capable of furnishing estimates for Ca2+- and Mg2+ binding constants in these systems. And because the method yields the binding enthalpy directly, ITC can provide a more detailed view of binding energetics than methods that rely on 45Ca2+ or fluorescent indicators. For several reasons, however, it is generally not possible to extract reliable binding parameters from single ITC experiments. Ca2+ affinity is often too high, and Mg2+ affinity is invariably too low. Moreover, least-squares minimization of multisite systems may not afford a unique fit because of strong parameter correlations. This chapter outlines a strategy for analyzing two-site systems that overcomes these obstacles. The method--which involves simultaneous, or global, least-squares analysis of direct and competitive ITC data--yields binding parameters for both Ca2+ and Mg2+. Application of the method is demonstrated for two systems. The S55D/E59D variant of rat alpha-parvalbumin, noteworthy for its elevated metal ion affinity, binds divalent ions noncooperatively and is amenable to analysis using an independent two-site model. On the other hand, Phl p 7, a pollen-specific EF hand protein from timothy grass, binds Ca2+ with positive cooperativity. Divalent ion-binding data for the protein must be analyzed using a two-site Adair model. PMID- 19289211 TI - Energetic profiling of protein folds. AB - Current protein classification methods treat high-resolution structures as static entities. However, experiments have well documented the dynamic nature of proteins. With knowledge that thermodynamic fluctuations around the high resolution structure contribute to a more physically accurate and biologically meaningful picture of a protein, the concept of a protein's energetic profile is introduced. It is demonstrated on a large scale that energetic profiles are both diagnostic of a protein fold and evolutionarily relevant. Development of Structural Thermodynamic Ensemble-based Protein Homology (STEPH), an algorithm that searches for local similarities between energetic profiles, constitutes a first step towards a long-term goal of our laboratory to integrate thermodynamic information into protein-fold classification approaches. PMID- 19289212 TI - Model membrane thermodynamics and lateral distribution of cholesterol: from experimental data to Monte Carlo simulation. AB - Thermodynamic analysis and Monte Carlo simulation techniques were used to study cholesterol-lipid interactions in lipid membranes. Experimental data, including the maximum solubility of cholesterol in lipid bilayers, the 1-to-1 displacement of cholesterol by ceramide, and the cholesterol chemical activity with cholesterol oxidase (COD), were systematically analyzed using thermodynamic principles. A conceptual model, the umbrella model, is presented to describe the key cholesterol-lipid interaction in lipid membranes. In a lipid membrane, nonpolar cholesterol relies on polar phospholipid headgroup coverage to avoid the unfavorable free energy of cholesterol contact with water. This coverage requirement leads to cholesterol's strong tendency not to clustering in a bilayer, its preferential association with large headgroup lipids with saturated acyl chains, and its competition with ceramide for large headgroup lipids. The umbrella model was parameterized into a form of multibody (i.e., nonpairwise) interaction for Monte Carlo simulation, and the measured chemical potentials of cholesterol agreed favorably with the predictions from the simulation. Under the right conditions, the multibody interactions can also lead to the formation of cholesterol superlattices. Also, an intrinsic thermodynamic connection between a jump in chemical potential and a regular distribution (RD) of membrane molecules was uncovered. This study shows that combining thermodynamics with computer simulation can be a productive approach for analyzing and interpreting complex experimental data, and thermodynamics can yield a predicting power in bioscience research. PMID- 19289213 TI - Thinking inside the box: designing, implementing, and interpreting thermodynamic cycles to dissect cooperativity in RNA and DNA folding. AB - Double and triple mutant thermodynamic cycles provide a means to dissect the cooperativity of RNA and DNA folding at both the secondary and tertiary structural levels through use of the thermodynamic box or cube. In this article, we describe three steps for applying thermodynamic cycles to nucleic acid folding, with considerations of both conceptual and experimental features. The first step is design of an appropriate system and development of hypotheses regarding which residues might interact. Next is implementing this design in terms of a tractable experimental strategy, with an emphasis on UV melting. The final step, and the one we emphasize the most, is interpreting mutant cycles in terms of coupling between specific residues in the RNA or DNA. Coupling free energy in the absence and presence of changes elsewhere in the molecule is discussed in terms of specific folding models, including stepwise folding and concerted changes. Last, we provide a practical section on the use of commercially available software (KaleidaGraph) to fit melting data, along with a consideration of error propagation. Along the way, specific examples are chosen from the literature to illustrate the methods. This article is intended to be accessible to the biochemist or biologist without extensive thermodynamics background. PMID- 19289215 TI - Extracting equilibrium constants from kinetically limited reacting systems. AB - It has been known for some time that slow kinetics will distort the shape of a reversible reaction boundary. Here we present a tutorial on direct boundary fitting of sedimentation velocity data for a monomer-dimer system that exhibits kinetic effects. Previous analysis of a monomer-dimer system suggested that rapid reaction behavior will persist until the relaxation time of the system exceeds 100 s (reviewed in Kegeles and Cann, 1978). Utilizing a kinetic integrator feature in Sedanal (Stafford and Sherwood, 2004), we can now fit for the k(off) values and measure the uncertainty at the 95% confidence interval. For the monomer-dimer system the range of well determined k(off) values is limited to 0.005 to 10(-5) s(-1) corresponding to relaxation times (at a loading concentration of the Kd) of approximately 70 to approximately 33,000 s. For shorter relaxation times the system is fast and only the equilibrium constant K but not k(off) can be uniquely determined. For longer relaxation times the system is irreversibly slow, and assuming the system was at initial equilibrium before the start of the run, only the equilibrium constant K but not k(off) can be uniquely determined. PMID- 19289214 TI - The thermodynamics of virus capsid assembly. AB - Virus capsid assembly is a critical step in the viral life cycle. The underlying basis of capsid stability is key to understanding this process. Capsid subunits interact with weak individual contact energies to form a globally stable icosahedral lattice; this structure is ideal for encapsidating the viral genome and host partners and protecting its contents upon secretion, yet the unique properties of its assembly and inter-subunit contacts allow the capsid to dissociate upon entering a new host cell. The stability of the capsid can be analyzed by treating capsid assembly as an equilibrium polymerization reaction, modified from the traditional polymer model to account for the fact that a separate nucleus is formed for each individual capsid. From the concentrations of reactants and products in an equilibrated assembly reaction, it is possible to extract the thermodynamic parameters of assembly for a wide array of icosahedral viruses using well-characterized biochemical and biophysical methods. In this chapter we describe this basic analysis and provide examples of thermodynamic assembly data for several different icosahedral viruses. These data provide new insights into the assembly mechanisms of spherical virus capsids, as well as into the biology of the viral life cycle. PMID- 19289216 TI - Facial trauma--who should provide care? AB - The management of facial trauma is key to the development and longevity of the specialty of Oral and Maxillofacial Surgery. Advances in material technology and surgical evolution have kept pace with increasingly complex fracture configurations. Soft tissue components of major craniofacial injuries are best managed in a multidisciplinary environment. This may be facilitated by a nominated team within specific trauma centres. Trauma management of the face is a continuum with both training implications and clinical governance issues. There is no uniformly accepted staging of severity, and no accepted quantifiable outcome measures. Until there are, craniofacial trauma management will remain anecdotal. PMID- 19289217 TI - Migraine and other primary headaches. Preface. PMID- 19289218 TI - The epidemiology, burden, and comorbidities of migraine. AB - We describe the epidemiology and comorbidities of migraine, which affects 12% of adults in occidental countries. Prevalence is three times higher in women, but 6% of men are affected, making it the most prevalent neurologic disorder in men. Although migraine is a remarkably common cause of temporary disability, many migraineurs have never consulted a physician for the problem. Many disorders are comorbid with migraine. For some such as depression, the association has been well described, but for others, the relationship has been recently suggested, such as in the case of clinical and subclinical vascular brain lesions and coronary heart disease. PMID- 19289219 TI - Pathophysiology of migraine. AB - Migraine is a common, disabling disorder of the central nervous system. The disorder has three key features. The tendency is largely inherited, the sufferer is sensitive to exogenous and endogenous triggers that very often involve challenges to normal homeostatic biology, and the attack phenotype, when severe, is the stereotypical migraine attack. The attack itself consists of an abnormal perception of otherwise normal circumstances, such as pain without evidence of primary nociceptive activation, and light and sound sensitivity without change in ambient stimuli. The disturbance in the brain is of the subcortical aminergic sensory modulatory systems, and probably includes brainstem, hypothalamic, and thalamic changes that produce the rich clinical presentation seen in practice. PMID- 19289220 TI - Transient neurologic dysfunction in migraine. AB - Neurologic symptoms are a prominent and often disturbing component of the migraine syndrome in many patients. Collectively termed "aura," migraine-related neurologic symptoms include visual, sensory, language, and motor disturbance. They occur in about one quarter of migraine patients, are classically transient, and are thought to occur as the result of cortical phenomena. Recently, motor symptoms previously included as a type of migraine aura have been reclassified as a component of hemiplegic migraine--a distinct migraine subtype. The tendency to aura is likely to be influenced by complex genetic and perhaps epigenetic factors. PMID- 19289221 TI - Vestibular migraine. AB - Vestibular migraine (VM) designates recurrent attacks of vertigo that are caused by migraine. VM presents with attacks of spontaneous or positional vertigo, lasting seconds to days, accompanied by migrainous symptoms. Because headache is often absent during acute attacks, other migrainous features have to be identified by thorough history taking. In contrast, vestibular testing serves mainly for the exclusion of other diagnoses. Treatment is targeted at the underlying migraine. PMID- 19289222 TI - Diagnostic testing for migraine and other primary headaches. AB - Most primary headaches can be diagnosed using the history and examination. Judicious use of neuroimaging and other testing, however, is indicated to distinguish primary headaches from the many secondary causes that may share similar features. This article evaluates the reasons for diagnostic testing and the use of neuroimaging, electroencephalography, lumbar puncture, and blood testing. The use of diagnostic testing in adults and children who have headaches and a normal neurologic examination, migraine, trigeminal autonomic cephalalgias, hemicrania continua, and new daily persistent headache are reviewed. PMID- 19289223 TI - Acute treatment of migraine. AB - The goals of acute treatment of migraine are cost-effective rapid relief, consistent reduction of disability, and reduced use of rescue medications. Key to acute treatment is matching medication to disability as a surrogate marker for disease severity with a stratified care approach. In the absence of vascular contraindications, triptans are first-line acute treatments for disabling migraine. Acute treatment success can be assessed with use of the paper tool, Migraine-ACT. Opioids and butalbital should be avoided in acute migraine treatment. Triptan type can be selected for patients by differences in speed or effect, formulation, and formulary tier. PMID- 19289224 TI - Preventive migraine treatment. AB - The pharmacologic treatment of migraine may be acute (abortive) or preventive (prophylactic), and patients with frequent severe headaches often require both approaches. Preventive therapy is used to try to reduce the frequency, duration, or severity of attacks. The preventive medications with the best-documented efficacy are amitriptyline, divalproex, topiramate, and the beta-blockers. Choice is made based on a drug's proven efficacy, the physician's informed belief about medications not yet evaluated in controlled trials, the drug's adverse events, the patient's preferences and headache profile, and the presence or absence of coexisting disorders. Because comorbid medical and psychologic illnesses are prevalent in patients who have migraine, one must consider comorbidity when choosing preventive drugs. Drug therapy may be beneficial for both disorders; however, it is also a potential confounder of optimal treatment of either. PMID- 19289225 TI - Behavioral medicine for migraine. AB - Behavioral medicine is based on the biopsychosocial theory that biological, psychological, and environmental factors all play significant roles in human functioning. This article reviews empirically supported and efficacious behavioral approaches to the treatment and management of migraine including cognitive behavioral therapy and biobehavioral training (ie, biofeedback, relaxation training, and stress management). These techniques have demonstrated efficacy when learned and practiced correctly and may be used individually or in conjunction with pharmacologic and other interventions. Data are also reviewed regarding patient education, support groups, psychological comorbidities, modifiable risk factors for headache progression, strategies for enhancing adherence and motivation, and strategies for effective medical communication. PMID- 19289226 TI - The face of chronic migraine: epidemiology, demographics, and treatment strategies. AB - Chronic daily headache (CDH) represents a therapeutic challenge for many clinicians. Treatment strategies should be aimed at correctly identifying the presence of CDH. In addition, an effective prophylactic regimen should be initiated; the presence of medication overuse must be addressed, and the offending medication being overused must be discontinued. Aside from analgesic overuse, other modifiable risk factors associated with the development of chronic migraine and CDH must be addressed including obesity and caffeine use and the effective management of comorbid conditions such as depression, anxiety, and sleep-related breathing disorders. PMID- 19289227 TI - Pediatric migraine. AB - Migraine headaches are common in children and adolescents, with a wide spectrum of clinical forms. The most frequent pattern in children is migraine without aura, characterized by attacks of frontal, pounding, nauseating headache lasting 1 to 72 hours. The spectrum of migraine with aura includes migraine with typical aura, hemiplegic migraine, and basilar-type migraine, all of which may manifest during early childhood and pose challenging diagnostic dilemmas. The periodic syndromes are a fascinating subset of migraine peculiar to extremely young children, which are viewed as "precursors" to more typical migraine and can be associated with frightening focal neurologic disturbances. Migraine treatment philosophy now embraces a balanced approach with biobehavioral interventions and acute and preventative pharmacologic measures. A growing body of controlled pediatric data is beginning to emerge regarding migraine treatment in children, lessening our dependence on extrapolated adult data. PMID- 19289228 TI - Migraine in women. AB - Of the nearly 32 million Americans with migraine, 24 million are women. It is a disorder affecting women throughout their lifetimes, from childhood and puberty through the postmenopausal years. In childhood, before puberty girls are afflicted with migraine at approximately the same rate as boys, but after puberty, there is an emerging female predominance. Estrogen plays a key role in this epidemiologic variation but is not the only factor. There are numerous times when hormonal influences have an impact on migraine and its pattern, including menarche, oral contraceptive use, pregnancy, perimenopause, and menopause. Hence practitioners treating women with migraine need to have a clear understanding of these special considerations. PMID- 19289230 TI - Tension-type headache. AB - The substantial societal and individual burdens associated with tension-type headache (TTH) constitute a previously overlooked major public health issue. TTH is prevalent, affecting up to 78% of the general population, and 3% suffer from chronic TTH. Pericranial myofascial nociception probably is important for the pathophysiology of episodic TTH, whereas sensitization of central nociceptive pathways seems responsible for the conversion of episodic to chronic TTH. Headache-related disability usually can be reduced by identification of trigger factors combined with nonpharmacologic and pharmacologic treatments, but effective treatment modalities are lacking. Benefits can be gained by development of specific and effective treatment strategies. PMID- 19289229 TI - The migraine association with cardiac anomalies, cardiovascular disease, and stroke. AB - Migraine is positively associated with cardio- and cerebrovascular disorders and with structural heart anomalies. Migraine is more prevalent among people with right-to-left shunt by means of patent foramen ovale, atrial septal defects, and pulmonary arteriovenous malformations and among those with altered cardiac anatomy, such as mitral valve prolapse, atrial septal aneurysm, and congenital heart disease. Meanwhile, migraine increases the risk for cardiovascular disease and stroke. Although several hypotheses exist, explanation for these associations is lacking. This article reviews data supporting the association of migraine with right-to-left shunt, structural heart anomalies, cardiovascular disease, and ischemic stroke. PMID- 19289231 TI - Trigeminal autonomic cephalalgias. AB - The trigeminal autonomic cephalalgias (TACs) are a group of primary headache syndromes all marked by headache and associated autonomic features. The TACs include cluster headache, paroxysmal hemicrania, hemicrania continua, and short lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing syndrome. Diagnosis is made after looking at headache frequency, duration, and accompanying symptoms. Each TAC has its own unique treatment modality, which is discussed in depth. PMID- 19289232 TI - Other primary headaches. AB - This article reviews "other primary headaches," a classification of the International Headache Society that includes primary stabbing headaches, primary cough headache, primary exertional headache, primary headache associated with sexual activity, hypnic headache, primary thunderclap headache, and new daily persistent headache. Clinicians should be aware that these headaches may be symptomatic to structural lesions and therefore usually require careful neuroimaging evaluation. PMID- 19289233 TI - What is needed to eradicate bovine tuberculosis successfully: An Ireland perspective. PMID- 19289234 TI - Local cytokine profiles in knee osteoarthritis: elevated synovial fluid interleukin-15 differentiates early from end-stage disease. AB - OBJECTIVE: Much of what is known about the inflammatory response in the synovial membrane (SM) of patients with osteoarthritis (OA) comes from studies of synovial tissues from end-stage disease. In this study, we sought to better characterize the inflammatory infiltrate in symptomatic patients with early signs of knee OA, and to determine how inflammatory cell populations relate to the pattern of cytokine and degradative enzyme production. METHODS: Study populations comprised patients with degenerative meniscal tears and early cartilage thinning undergoing arthroscopic procedures (early OA) and patients undergoing total knee replacement for end-stage OA. Quantitative real-time polymerase chain reaction (PCR) was used to measure expression of SM cytokines and enzymes implicated in the pathogenesis of inflammatory arthritis and OA, as well as cell lineage-specific markers. We quantified synovial fluid (SF) cytokines and enzymes by enzyme-linked immunosorbent assay (ELISA) and SM cell populations by immunohistochemistry. RESULTS: We found increased levels of SF interleukin-15 (IL-15) protein in the early knee OA patients when compared to end-stage OA. Both SF IL-15 protein and numbers of CD8 cells within SM correlated with matrix metalloproteinase-1 (MMP-1) and three levels. TNF-alpha, IL-6 and IL-21 were also detectable in the SF of the majority of patients, and IL-15 levels were associated with IL-6 levels. CONCLUSION: IL-15 is elevated in early knee OA, suggesting activation of an innate immune response in the SM. The association of IL-15 expression with CD8 transcripts and MMPs implicates this cytokine in OA pathogenesis and as a candidate therapeutic target. PMID- 19289235 TI - Chondroitin sulphate: an effective joint lubricant? AB - OBJECTIVE: The effect of chondroitin sulphate (CS) treatment on the friction and deformation characteristics of native and glycosaminoglycan (GAG) deficient articular cartilage was investigated. METHODS: Friction tests were conducted at 0.4 MPa load, in Static and Dynamic models, to determine the startup coefficient of friction (COF) and dynamic COF, respectively. Native cartilage: For each cartilage pin and plate couple, the COF was determined under three consecutive tests - (1) baseline COF in PBS (2) COF in CS lubricant and (3) COF again in PBS, after 24h CS treatment. GAG deficient cartilage: For each cartilage pin and plate couple, the baseline COF was determined in PBS initially and again following enzymatic treatment to deplete GAGs. The specimens were then soaked in CS solution for 24h and the COF determined again in PBS. In a similar manner, friction tests were replaced with indentation tests to study the deformation of the tissue. RESULTS: CS at 50mg/ml significantly lowered the startup COF of native cartilage both as a lubricant and a treatment solution. In the dynamic model, where the fluid load support is sustained at a high level, CS failed to have any effect on the COF of native cartilage. GAG depletion raised the friction and deformation levels of cartilage, and subsequent CS treatment failed to lower them to their native levels. CONCLUSION: CS proved to be an effective lubricant for cartilage under mixed-mode lubrication conditions. However, supplemental CS that diffused into the specimens had no influence on the fluid load support of cartilage. PMID- 19289236 TI - When stereotype knowledge prevents retrieval-induced forgetting. AB - In the present study, we examined the effects of stereotypes and the typicality of traits in retrieval-induced forgetting (RIF). The participants in the experiment practiced half of the high-typicality, low-typicality or control traits associated with the name of a stereotype (athlete, scientist) or with the name of a person (Mikel, Jon). With the person name the high-typicality, low typicality or control traits produced no evidence of stereotype activation. The traits were processed as independent features and generated RIF both immediately and at a one-week interval. Stereotype activation during encoding facilitated the integration of high-typicality traits, avoiding RIF in immediate and one-week recall. Moreover, both high-typicality and low-typicality traits benefited from stereotypes in one-week recognition. These findings show that previous knowledge contributes to integrate stereotypic traits that would otherwise compete for retrieval, thus producing RIF. PMID- 19289237 TI - Children's multiplicative transformations of discrete and continuous quantities. AB - Recent studies have documented an evolutionarily primitive, early emerging cognitive system for the mental representation of numerical quantity (the analog magnitude system). Studies with nonhuman primates, human infants, and preschoolers have shown this system to support computations of numerical ordering, addition, and subtraction involving whole number concepts prior to arithmetic training. Here we report evidence that this system supports children's predictions about the outcomes of halving and perhaps also doubling transformations. A total of 138 kindergartners and first graders were asked to reason about the quantity resulting from the doubling or halving of an initial numerosity (of a set of dots) or an initial length (of a bar). Controls for dot size, total dot area, and dot density ensured that children were responding to the number of dots in the arrays. Prior to formal instruction in symbolic multiplication, division, or rational number, halving (and perhaps doubling) computations appear to be deployed over discrete and possibly continuous quantities. The ability to apply simple multiplicative transformations to analog magnitude representations of quantity may form a part of the toolkit that children use to construct later concepts of rational number. PMID- 19289238 TI - Coordinating the effects of multiple variables: a skill fundamental to scientific thinking. AB - The skill of predicting outcomes based on simultaneous effects of multiple factors was examined. Over five sessions, 91 sixth graders engaged this task either individually or in pairs and either preceded or followed by six sessions on the more widely studied inquiry task that requires designing and interpreting experiments to identify individual effects. Final assessment, while indicating a high level of mastery on the inquiry task, showed progress but continuing conceptual challenges on the multivariable prediction task having to do with understanding of variables, variable levels, and consistency of a variable's operation across occasions. Task order had a significant but limited effect, and social collaboration conferred only a temporary benefit that disappeared in a final individual assessment. In a follow-up study, the lack of effect of social collaboration was confirmed, as was that of feedback on incorrect answers. Although fundamental to science, the concept that variables operate jointly and, under equivalent conditions, consistently across occasions is one that children appear to acquire only gradually and, therefore, one that cannot be assumed to be in place. PMID- 19289239 TI - Math in actions: actor mode reveals the true arithmetic abilities of French speaking 2-year-olds in a magic task. AB - Our previous studies provide some evidence of between-language effects on arithmetic performance in 2-year-olds. French-speaking children were especially biased by the use of the word un as a cardinal value and as an article in the singular/plural opposition (1 vs. the set 2, 3, ...). Here we evaluated the ability of a new action-based assessment method to avoid this bias. A total of 80 French-speaking 2- and 3-year-olds were confronted with impossible (1+1=1 or 1+1=3) and possible (1+1=2) addition problems that triggered the bias. The problems were either presented to the children by the experimenter (onlooker mode) or realized by themselves (actor mode). The 2-year-olds performed better in the actor mode than in the onlooker mode. A subtraction control with no language ambiguity (2-1=2 or 1) was conducted with 80 other children; both modes elicited comparable performances regardless of age. These data indicate that the actor mode is effective for assessing arithmetic ability in French-speaking 2-year olds. PMID- 19289240 TI - The acute antipanic and anxiolytic activity of aerobic exercise in patients with panic disorder and healthy control subjects. AB - Regular physical activity is anxiolytic in both healthy subjects and patients with panic disorder. In contrast, acute exercise may induce acute panic attacks or increase subjective anxiety in patients with panic disorder more than in other people. The effects of quiet rest or an aerobic treadmill exercise (30 min at an intensity of 70% of the maximal oxygen uptake, VO2max) on cholecystokinin tetrapeptide (CCK-4) induced panic attacks were studied in a crossover design in 12 patients with panic disorder and 12 matched healthy subjects. The effects of CCK-4 (25 microg in patients and 50 microg in control subjects) were measured with the Acute Panic Inventory (API) score, comparing panic attack frequencies, total score, and subscores for anxiety and somatic symptoms. CCK-4-induced panic attacks were less frequent after prior exercise: they occurred in 15 (62.5%) subjects after rest (9 patients and 6 control subjects), but only 5 (20.8%) subjects after exercise (4 patients and 1 control subject). In both conditions, CCK-4 administration induced a significant increase in the total API score and the anxiety and somatic symptoms subsores. However, compared to prior rest, exercise resulted in a significantly reduced CCK-4-induced increase of the total API score and the anxiety subscore. In patients with panic disorder exercise increased the total API score and the somatic symptoms subscale but not the anxiety subscore. Patients with panic disorder showed increased somatic but not anxiety symptoms after an acute bout of exercise. Severity of CCK-4-induced panic and anxiety, on the other hand was reduced by exercise. These findings suggest that in addition to exercise training an acute bout of exercise may be used to reduce anxiety and panic attack frequency and intensity in panic disorder patients. PMID- 19289241 TI - Improved detection of incipient vascular changes by a biotechnological platform combining post mortem MRI in situ with neuropathology. AB - The histopathological counterpart of white matter hyperintensities is a matter of debate. Methodological and ethical limitations have prevented this question to be elucidated. We want to introduce a protocol applying state-of-the-art methods in order to solve fundamental questions regarding the neuroimaging-neuropathological uncertainties comprising the most common white matter hyperintensities [WMHs] seen in aging. By this protocol, the correlation between signal features in in situ, post mortem MRI-derived methods, including DTI and MTR and quantitative and qualitative histopathology can be investigated. We are mainly interested in determining the precise neuroanatomical substrate of incipient WMHs. A major issue in this protocol is the exact co-registration of small lesion in a tridimensional coordinate system that compensates tissue deformations after histological processing. The protocol is based on four principles: post mortem MRI in situ performed in a short post mortem interval, minimal brain deformation during processing, thick serial histological sections and computer-assisted 3D reconstruction of the histological sections. This protocol will greatly facilitate a systematic study of the location, pathogenesis, clinical impact, prognosis and prevention of WMHs. PMID- 19289242 TI - Effects of multisensory stimulation on cognition, depression and anxiety levels of mildly-affected Alzheimer's patients. AB - AIMS: The purpose of this study was to investigate and assess the effects of musical therapy, painting inanimate-animate object pictures, and orientation to time-place-person interventions on the cognitive state, depression, and anxiety levels of mildly-affected Alzheimer's patients. METHODS: The study using a quasi experimental design was conducted with 27 mildly-affected Alzheimer's patients. The effects of the multisensory stimulation were evaluated with the "Mini Mental State Examination," the "Geriatric Depression Scale," and the "Beck Anxiety Scale." All of these were administered one day prior to beginning the study, immediately after its completion, and three weeks thereafter. RESULTS: A significant negative correlation was determined to exist between the MMSE depression scores and MMSE-anxiety scores; the correlation between the depression anxiety scores, on the other hand, had a positive significance. The shifts over time in the MMSE, depression and anxiety scores were significant. CONCLUSION: The primary conclusion of the study is that the multisensory stimulation method applied to mildly-affected Alzheimer's patients had a positive effect on their cognitive state, depression, and anxiety, and that this effect continued for three weeks following completion of the study intervention, with a tendency to decline progressively. PMID- 19289243 TI - Determination of microbial volatile organic compounds (MVOCs) by passive sampling onto charcoal sorbents. AB - Passive sampling combined with GC-MS was applied for the detection of microbial volatile organic compounds (MVOCs) emitted by mould cultures. The fungi species Penicillium expansum, Penicillium chrysogenum, Penicillium brevicompactum, Aspergillus fumigatus, Aspergillus versicolor, Aspergillus niger, and Cladosporium cladosporoides were cultivated on dichloran glycerol agar. The emitted volatiles were collected by charcoal-based passive diffusion monitors during the first four weeks of growth. Several alcohols, mainly 2-methyl-1 propanol and 3-methyl-1-butanol, alkenes, ketones, ethers and esters could be identified and quantified after external calibration. Species-specific compounds like 1,3-dimethoxybenzene for A. versicolor could be proposed. Associations between the concentrations of some of the identified MVOCs (e.g. methylfurans and dimethyldisulfide) and indoor mould exposure in dwellings were found in a field study comprising more than 500 dwellings in Leipzig/Germany. It could be concluded that passive sampling onto charcoal adsorbents for the determination of MVOCs is particularly suitable in epidemiological studies with a high number of participants. PMID- 19289244 TI - Simultaneous autotrophic biodegradation of H2S and NH3 in a biotrickling filter. AB - An efficient co-treatment of H(2)S and NH(3) was achieved in a horizontal biotrickling filter (HBTF) inoculated by autotrophic nitrifying and sulfide oxidizing microbial consortia and packed with exhausted carbon. NH(3) and H(2)S were removed completely at the beginning of operation, in terms of nitrate and sulfate mainly by autotrophic biodegradation, respectively. Under the steady state, removal efficiency (RE) of H(2)S and NH(3) was investigated with different gas retention times (GRT=4-20s), inlet concentrations (20-100 ppmv) and inlet loadings. At GRT>4s, above 98% of NH(3) was removed while 95% RE of H(2)S was reached at GRT>8s. The critical elimination capacity (at 99% RE) was attained at 44 g NH(3)m(-3)h(-1) and 36 g H(2)Sm(-3)h(-1), and no interaction between NH(3) and H(2)S treatment was found at low loadings. Furthermore, the experiment at high loadings of NH(3) and H(2)S was purposely designed to evaluate the potential inhibition of each other. The metabolic products of N and S bearing species in recycling liquid were analyzed to explore the removal mechanisms. Compared with previous studies, the present filter showed a better removal capacity of NH(3) and H(2)S. Overall, the results indicated that the HBTF was highly efficient and cost-effective for simultaneous autotrophic biodegradation of NH(3) and H(2)S. PMID- 19289245 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) plasma concentrations in residents of Paritutu, New Zealand: evidence of historical exposure. AB - An assessment of community exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was undertaken in Paritutu, New Zealand. The suburb lies adjacent to an agrichemical facility that produced 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), between 1962 and 1987. Soil TCDD measurements from 73 nearby addresses demonstrated a pattern of TCDD deposition consistent with an aerial plume following the prevailing local wind patterns and the agrichemical plant as the point source. Blood samples were taken from 52 volunteers having lived for three or more years in Paritutu between 1962 and 1987. Candidate selection focused primarily on individuals who were most likely to show elevated TCDD blood lipid levels when compared to age and gender stratified national average blood concentrations, and secondarily on individuals that provided additional information about specific exposure periods, potential exposures of younger age groups, and specific dietary patterns. A multipathway exposure model was used to estimate serum TCDD levels in each participant. Age and gender-specific TCDD elimination kinetics were also considered. Historical TCDD environmental concentrations were back-calculated from soil concentrations at each residence assuming TCDD releases occurred pre-dominantly between 1962 and 1975. Serum was analysed for chlorinated dibenzodioxins and dibenzofurans, and a subset was analysed for dioxin-like polychlorinated biphenyls. TCDD in serum lipid exceeded two standard deviations above national background levels for 14 participants, and 3 standard deviations for 10 participants. The highest TCDD lipid concentration was 33.3 ng/kg-lipid, or 11 times higher than the comparative 1997 national average. Elevated TCDD concentrations were observed primarily, but not exclusively, in the older study participants who had been in residence in Paritutu before 1968. The study demonstrated TCDD exposure in this community, occurring most likely through the aerial route, and most probably from fugitive emissions during manufacture. PMID- 19289246 TI - Influence of fluorocarbon flat-membrane hydrophobicity on carbon dioxide recovery. AB - The influence of hydrophobicity in flat-plate porous poly(vinylidene fluoride) (PVDF) and expended polytetrafluoroethylene (PTFE) membranes on CO(2) recovery using aqueous solutions of piperazine (PZ) and alkanolamine is examined. Experiments were conducted at various gas flow rates, liquid flow rates, and absorbent concentrations. The CO(2) absorption flux increased with increasing gas flow rates and absorbent concentrations. When using 2-amino-2-methyl-1-propanol (AMP) or AMP+PZ aqueous solution as absorbent, this process was dominantly governed by gas film layer diffusion and membrane diffusion. The diffusion resistance of the membrane phase was only important when using N methyldiethanolamine as the sole absorbent. The water contact angle increased initially with increasing plasma working power and reached at steady state value of 155 degrees beyond 100 W. The elemental fluorine-to-carbon ratio (F/C) and water contact angle of the PVDF membrane increased with increasing treatment time and reached a plateau after 5min of CH(4) plasma (100 W). Increases in the CO(2) absorption fluxes of 7% and 17% were observed for plasma-treated PVDF membranes in comparison to non-treated PVDF and PTFE, respectively, when using 1M AMP as absorbent. The membrane mass transfer coefficient, k(m), for plasma-treated PVDF membranes increased from 2.1 x 10(-4) to 2.5 x 10(-4)ms(-1). Membrane durability was greatly improved by CF(4) plasma treatment (100 W/5 min) and comparable to that of PTFE membranes. PMID- 19289247 TI - Application of semipermeable membrane device (SPMD) to assess air genotoxicity in an occupational environment. AB - Semipermeable membrane device (SPMD) is a passive sampler that sequesters lipophilic contaminants, mimicking the bioconcentration in the fatty tissue of organisms. This study was designed to assess the use of SPMD and biological tests (Comet assay and Ames test) for air monitoring. For this purpose an occupational environment with expected polycyclic aromatic hydrocarbons (PAHs) contamination (coke plant) was selected for a case study. The SPMDs were deployed in five occupational contaminated sites and in a control site. The SPMD dialysates were chemically analysed and examined for in vitro DNA-damaging activity in human cells (Jurkat) by Comet assay and for mutagenicity with the Ames test (TA98 strain, w/o S9). Total suspended particulates were also collected and analysed (GC-MS). No biological effect of SPMD extract was revealed in the control site. On the other hand, air samples collected with SPMDs within the coke plant showed variable degrees of genotoxic and mutagenic activity. The highest effects were associated with the highest PAH level recovered in the SPMDs extracts and in particulate samples. Results obtained support the sensitivity of biological tests associated to SPMD sampling for evaluating the health risk of potentially contaminated work environments highlighting the usefulness of SPMDs for environmental air quality monitoring. PMID- 19289248 TI - An antimicrobial modified silicone peritoneal catheter with activity against both Gram-positive and Gram-negative bacteria. AB - Peritonitis, exit site and tunnel infections are serious complications of peritoneal dialysis (CAPD), which may lead to catheter loss, despite measures taken to reduce the infection rate. Catheters coated with antimicrobials have shown only short-term activity. We have developed a process for conferring broad spectrum, long-duration antimicrobial activity on CAPD catheters while reducing the risk of resistance. Catheters were processed using an impregnation method. Three agents were used: rifampicin, triclosan and trimethoprim. Tests included establishing the duration of activity before test bacteria became resistant, the ability to kill 100% of attached bacteria in a predetermined time (tK100), and ability to withstand multiple high-count challenges until failure in a flow model. Antimicrobial activity continued until it was stopped at 280 days and the agents prevented the emergence of resistant strains. tK100 results showed a >99.9% reduction of attached bacteria. The flow model showed no colonization when repeatedly challenged with high loads of meticillin-resistant Staphylococcus aureus or Escherichia coli for approximately 90 days. The antimicrobial catheter material showed prolonged activity against common CAPD pathogens and promises to reduce clinical CAPD infection. PMID- 19289249 TI - Natural time course of myocardial infarction at delayed enhancement magnetic resonance. AB - The case reported, who suffered prolonged coronary occlusion, was evaluated by repeated MRI images in the acute, subacute and chronic phase. Scans showed hemorrhagic myocardial infarction and massive endothelial injury and its evolution over time. PMID- 19289250 TI - Skeletal muscle proteins oxidation in chronic right heart failure in rats: can different beta-blockers prevent it to the same degree? AB - BACKGROUND: Skeletal muscle atrophy and decreased expression of slow fibers contribute to exercise capacity limitation in Chronic Heart Failure (CHF). Pro inflammatory cytokines and free radicals worsen muscle damage. In CHF sarcomeric proteins are oxidized with reduction of muscle twitch efficiency, and VO(2)-max. Beta-blockers with anti-oxidative capacity such as carvedilol have been shown to prevent contractile protein oxidation in CHF rats. Recently a new class of beta blockers with NO donor activity has been introduced and approved for the treatment of CHF. Since a clinical clear superiority of a beta-blocker has never been shown, we compared nebivolol, that possesses NO donor activity, with bisoprolol, looking at possible differences in skeletal muscle that may have an impact on muscle function and exercise capacity in humans. We therefore studied skeletal muscle apoptosis and wastage, sarcomeric protein composition and oxidation, and muscle efficiency. METHODS AND RESULTS: In the monocrotaline rat model of CHF we compared nebivolol a beta-blocker with vasodilative properties mediated by NO production, with bisoprolol. Nebivolol prevented protein oxidation, while bisoprolol did it only partially, as demonstrated by the oxyblot analysis (Oxy/RP values) (0.90+/-0.14 Controls.; 1.7+/-0.14 CHF; 1.1+/-0.05 bisoprolol; 0.82+/-0.17 nebivolol low; 0.62+/-0.10 nebivolol high). Only nebivolol improved twitch force production and relaxation. Nebivolol prevented fibers shift towards fast isoforms, atrophy, decreased apoptosis and sphingosine levels. CONCLUSIONS: Nebivolol seems better than bisoprolol in CHF by decreasing apoptosis and cytokines induced muscle wastage, preventing fibers shift and protein oxidation. Nebivolol by stimulating NO generation may have prevented protein oxidation. It could be speculated that ROS release, pro-inflammatory cytokines production and NF-kappa-B activation may play a key role. These positive changes could produce a favorable impact on exercise capacity in man. PMID- 19289251 TI - Seasonal fluxes and temperature-dependent accumulation of persistent organic pollutants in lakes: the role of internal biogeochemical cycling. AB - A dynamic flux model for lakes taking into account the interactions between atmospheric depositional and biogeochemical processes (BIODEP model) was used to assess the importance of internal cycling and biogeochemical processes with respect to accumulation of four different polychlorinated biphenyls (PCBs) (congeners 28, 52, 101 and 153) in Lake Redo, a high-altitude lake in the Spanish Pyrenees. To investigate the effect of temperature on sediment accumulation, the model was run at different temperatures and corresponding sediment inventories were plotted vs. reciprocal temperature. In line with measurements from a previous study looking at sediment inventories of 19 European high-altitude lakes (MOLAR study), we found a stronger temperature dependence of lake sediment concentrations for the less volatile/less soluble PCBs. Seasonal and annual mass balances were investigated and highlighted the importance of internal lake processes controlling the differences in sediment accumulation for the different PCB congeners. For example, the higher temperature dependence of sediment inventories for the high chlorinated congeners is due to the fast dynamics of water-to-sediment transport in comparison to atmospheric deposition processes. PMID- 19289252 TI - "Nuts, schiz, psycho": an exploration of young homeless people's perceptions and dilemmas of defining mental health. AB - Through our research we explore the term 'mental health' as articulated by a group of young people living in homeless shelters utilising staff in a mental health service. This mental health service was offered in 5 large geographical areas (urban, semi-urban and rural) in England to 18 homeless shelters and we interviewed 25 homeless young people, 5 Mental Health Coordinators and 12 homeless shelter staff. Using discourse analysis of semi-structured interviews, we investigate the ideological dilemmas presented by the young people. They report negative and stigmatising descriptions of mental health despite their involvement with a mental health service. Four key interpretative repertoires are identified; denial of problems, mental health as negative, the need to talk, and challenging prejudice. It is concluded that the term 'mental health', which appears in the title of the service (of which they are clients), presents barriers for usage but works to challenge prejudice and educate young people. Discussion of the implications of naming services and the importance of shared meanings are considered. PMID- 19289253 TI - Does health predict the reporting of racial discrimination or do reports of discrimination predict health? Findings from the National Longitudinal Study of Youth. AB - Racial discrimination may contribute to diminished well-being, possibly through stress and restricted economic advancement. Our study examines whether reports of racial discrimination predict health problems, and whether health problems predict the reporting of racial discrimination. Data come from years 1979 to 1983 of the US National Longitudinal Study of Youth, focusing on respondents of Black (n=1851), Hispanic (n=1170), White (n=3450) and other (n=1387) descent. Our analyses indicate that reports of racial discrimination in seeking employment predict health-related work limitations, although these limitations develop over time, and not immediately. We also find that reports of discrimination at two time-points appear more strongly related to health-related work limitations than reports at one time-point. A key finding is that these limitations do not predict the subsequent reporting of racial discrimination in seeking employment. These findings inform our knowledge of the temporal ordering of racial discrimination in seeking employment and health-related work conditions among young adults. The findings also indicate that future research should carefully attend to the patterns and timing of discrimination. PMID- 19289254 TI - Cyclooxygenase-2/prostaglandin E2 pathway mediates icariside II induced apoptosis in human PC-3 prostate cancer cells. AB - Icariside II (IS) isolated from the roots of Epimedium koreanum Nakai was known to have antioxidant activity and inhibit melanogenesis and hypoxia inducible factor. We report here for the first time that IS induces apoptosis through its anti-inflammatory effects in PC-3 prostate cancer cells. IS exerted cytotoxicity against PC-3 cells with IC(50) of approximately 20 microM. IS suppressed both constitutive and arachidonic acid (AA)-induced cyclooxygenase-2 (COX-2) expression as well as reduced prostaglandin E2 (PGE2) levels in PC-3 cells even at a low concentrations (5 and 10 microM). Additionally, IS increased sub G1 apoptotic portion and exhibited terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL)-positive apoptotic bodies in PC-3 cells at higher concentrations (20 and 40 microM). Furthermore, IS attenuated the mitochondrial membrane potential, released cytochrome C into cytosol, activated caspase-9, -8, and -3 expressions and cleaved poly (ADP-ribose) polymerase (PARP) in PC-3 cells. Consistently, COX-2, inducible NO synthase (iNOS), and vascular endothelial growth factor (VEGF) expressions were suppressed while in parallel inducing apoptosis in hormone-independent prostate carcinoma cells PC-3. Moreover, exogeneous PGE2 inhibited IS induced PARP cleavage in PC-3 cells and also knockdown of COX-2 by siRNA potentiated IS induced PARP cleavage, thereby implicating the critical role of COX-2 pathway in IS induced apoptosis. Taken together, these findings demonstrate that IS initiates the inhibition of COX 2/PGE(2) pathway and then induces apoptosis mainly via mitochondrial dependent pathway in PC-3 prostate cancer cells as a potent cancer chemotherapeutic agent. PMID- 19289255 TI - Isoform-specific histone deacetylase inhibitors: the next step? AB - Histone deacetylases (HDACs) have emerged as attractive drug targets, particularly for neoplastic indications. This large family is divided into four classes, of which three consist of zinc-dependent enzymes, and inhibitors of these are the subject of this review. Currently, there are several inhibitors advancing through clinical trials, all of which inhibit multiple isoforms of these three classes. While promising, these compounds have exhibited toxicities in the clinic that might limit their potential, particularly in solid tumors. It may be possible to reduce some of the toxicity by specifically targeting only the isoform(s) involved in maintaining that particular tumor and spare other isoforms that are uninvolved or even beneficial. This review examines the selectivity and toxicity of HDAC inhibitors currently in clinic, comparing pan-HDAC inhibitors to Class I selective compounds. The rationale for isoform-specific inhibitors is examined. The current status of isoform-specific inhibitor development is analyzed, especially inhibitors of HDAC1, 2, 4 and 8 enzymes, and the potential clinical utility of these compounds is discussed. PMID- 19289256 TI - Differentiation of pain ratings in combat-related posttraumatic stress disorder. AB - Although posttraumatic stress disorder (PTSD) is associated with chronic pain, preliminary evidence suggests reduced experimental pain sensitivity in this disorder. The questions addressed in the present study were whether pain perception would also be reduced in PTSD patients who are not suffering from chronic pain symptoms, and whether a reduction in pain sensitivity would also be present in combat veterans who did not develop PTSD. For this, we determined thermal detection and pain thresholds in 10 male combat-related PTSD patients, 10 combat control subjects (no PTSD) and 10 healthy controls without combat experience. All subjects were pain free. First, we measured thermal sensory thresholds with ramped heat and cold stimuli using the method of limits. Ramped thermal sensory stimulation revealed no deficits for the detection of (non noxious) f2.1thermal stimuli between groups. In contrast, heat and cold pain thresholds in both combat groups (PTSD and combat controls) were significantly increased compared to healthy controls. However, these stimuli could not distinguish between the two groups due to ceiling effects. When using longer lasting heat stimulation at different temperatures (30s duration; method of fixed stimuli), we found significantly lower frequency of pain reports in PTSD patients compared with both combat and healthy controls, as well as significantly lower pain ratings. Our results suggest an association of PTSD with reduced pain sensitivity, which could be related to PTSD-related (neuro-)psychological alterations or to a pre-existing risk factor for the disorder. PMID- 19289257 TI - The therapeutic strategies of enhancing adiponectin and lowering leptin may be benefit to controlling atrial fibrillation. PMID- 19289258 TI - Expression of FXYD-3 is an independent prognostic factor in rectal cancer patients with preoperative radiotherapy. AB - PURPOSE: FXYD-3 (MAT-8) is overexpressed in several types of cancers; however, its clinical relevance in rectal cancers has not been studied. Therefore, we examined FXYD-3 expression in rectal cancers from the patients who participated in a Swedish clinical trial of preoperative radiotherapy (RT) to determine whether FXYD-3 was overexpressed in rectal cancers and correlated with RT, survival, and other clinicopathologic variables. METHODS AND MATERIALS: The study included 140 rectal cancer patients who participated in a clinical trial of preoperative RT, 65 with and 75 without RT before surgery. FXYD-3 expression was immunohistochemically examined in distant (n = 70) and adjacent (n = 101) normal mucosa, primary tumors (n = 140), and lymph node metastasis (n = 36). RESULTS: In the whole cohort, strong FXYD-3 expression was correlated with infiltrative tumor growth (p = 0.02). In the RT group, strong FXYD-3 expression alone (p = 0 .02) or combined with phosphatase of regenerating liver was associated with an unfavorable prognosis (p = 0.02), independent of both TNM stage and tumor differentiation. In tumors with strong FXYD-3 expression, there was less tumor necrosis (p = 0.02) and a trend toward increased incidence of distant metastasis (p = 0.08) after RT. None of these effects was seen in the non-RT group. FXYD-3 expression in the primary tumors tended to be increased compared with normal mucosa regardless of RT. CONCLUSION: FXYD-3 expression was a prognostic factor independent of tumor stage and differentiation in patients receiving preoperative RT for rectal cancer. PMID- 19289259 TI - Extrapolation of normal tissue complication probability for different fractionations in liver irradiation. AB - PURPOSE: The ability to predict normal tissue complication probability (NTCP) is essential for NTCP-based treatment planning. The purpose of this work is to estimate the Lyman NTCP model parameters for liver irradiation from published clinical data of different fractionation regimens. A new expression of normalized total dose (NTD) is proposed to convert NTCP data between different treatment schemes. METHOD AND MATERIALS: The NTCP data of radiation- induced liver disease (RILD) from external beam radiation therapy for primary liver cancer patients were selected for analysis. The data were collected from 4 institutions for tumor sizes in the range of of 8-10 cm. The dose per fraction ranged from 1.5 Gy to 6 Gy. A modified linear-quadratic model with two components corresponding to radiosensitive and radioresistant cells in the normal liver tissue was proposed to understand the new NTD formalism. RESULTS: There are five parameters in the model: TD(50), m, n, alpha/beta and f. With two parameters n and alpha/beta fixed to be 1.0 and 2.0 Gy, respectively, the extracted parameters from the fitting are TD(50)(1) = 40.3 +/- 8.4Gy, m =0.36 +/- 0.09, f = 0.156 +/- 0.074 Gy and TD(50)(1) = 23.9 +/- 5.3Gy, m = 0.41 +/- 0.15, f = 0.0 +/- 0.04 Gy for patients with liver cirrhosis scores of Child-Pugh A and Child-Pugh B, respectively. The fitting results showed that the liver cirrhosis score significantly affects fractional dose dependence of NTD. CONCLUSION: The Lyman parameters generated presently and the new form of NTD may be used to predict NTCP for treatment planning of innovative liver irradiation with different fractionations, such as hypofractioned stereotactic body radiation therapy. PMID- 19289260 TI - Repeat computed tomography simulation to assess lumpectomy cavity volume during whole-breast irradiation. AB - PURPOSE: To determine whether the lumpectomy cavity (LPC) decreases in volume during whole-breast radiotherapy (RT) and what factors influence the decrease. PATIENTS AND METHODS: Forty-three women with 44 breast lesions were prospectively enrolled. Eligible patients underwent lumpectomy followed by a CT simulation (CT1) within 60 days of surgery. Patients were treated to the entire breast to a dose of 45-50.4 Gy. After 21-23 treatments, a second planning CT simulation (CT2) was done. The LPC was contoured on CT2, and the volumes (LCV) were compared between CT1 and CT2. RESULTS: The median LCV on CT1 and CT2 was 38.2 cm(3) and 21.7 cm(3), respectively. The median percent change and volume decrease between CT1 and CT2 was -32.0% and 11.2 cm(3), respectively (n = 44). The LCV decreased in 38 of 44 patients (86%). There was a significant correlation between initial LCV and decrease in volume (p = 0.001) and initial LCV and percent decrease in volume (p < 0.001). There was no correlation between time from surgery to CT1, to start of RT, or to CT2 and change in volume. CONCLUSIONS: Patients who undergo lumpectomy almost always have a decrease in their LCV during whole-breast RT. There was a correlation between the initial LCV and decrease in volume on repeat CT simulation. Evaluating patients for this change can potentially lead to decreased doses of radiation to the remaining breast and other critical structures when delivering a small-field boost. Repeat CT simulation should be considered in patients with larger cavities or cavities near critical structures. PMID- 19289261 TI - lymph node ratio as a prognostic factor in patients with stage III rectal cancer treated with total mesorectal excision followed by chemoradiotherapy. AB - PURPOSE: To investigate the prognostic impact of lymph node ratio (LNR) on survival in the patients with Stage III rectal cancer. METHODS AND MATERIALS: We retrospectively reviewed the data of 421 consecutive patients who underwent total mesorectal excision followed by chemoradiotherapy for rectal cancer from 1996 to 2006. The 232 patients with positive lymph nodes (LNs) were divided into four groups according to LNR quartiles: LNR 0.4 (n = 60). The association between LNR and survival was evaluated by the Kaplan-Meier method and multivariate analysis with covariates of prognostic significance in univariate analysis. RESULTS: The median numbers of examined and positive LNs were 17 and 3, respectively, and the median LNR was 0.20 (range, 0.03-1). There was a strong correlation between the number of positive LNs and LNR (r = 0.724, p < 0.001). After a median follow-up of 53 months (range, 9-138 months), the actuarial overall survival and disease-free survival rates at 5 years were 69% and 56%, respectively. The 5-year survival rate decreased as LNR increased (0.4, 50%; p < 0.001). Lymph node ratio was also a significant prognostic factor on Cox regression analysis (0.4, HR = 3.7, p = 0.005). Lymph node ratio had a prognostic effect on overall survival in subgroups of patients with N1 (p = 0.032) and N2 (p = 0.034) tumors. CONCLUSION: Lymph node ratio was the most significant predictor of survival in the patients with Stage III rectal cancer who had undergone postoperative chemoradiation. PMID- 19289262 TI - Patterns of care and locoregional treatment outcomes in older esophageal cancer patients: The SEER-Medicare Cohort. AB - PURPOSE: Optimal management of elderly patients with nonmetastatic esophageal cancer is unclear. Outcomes data after locoregional treatment are lacking for this group. METHODS: We assessed outcomes associated with standard locoregional treatments in 2,626 patients (age > 65 years) from the Surveillance Epidemiology and End Results (SEER)-Medicare cohort diagnosed with nonmetastatic esophageal cancer from 1992 to 2002. In patients treated with radiotherapy alone (RT), surgery alone (S), chemoradiotherapy (CRT), or preoperative chemotherapy followed by surgery (CRT + S), overall and disease-free survival were compared using proportional hazards regression. Postoperative complications were compared using logistic regression. RESULTS: Mean age was 76 +/- 6 years. Seven percent underwent CRT + S, 39% CRT, 30% S, and 24% RT. One-year survival was 68% (CRT + S), 52% (CRT), 53% (S), and 16% (RT), respectively (p < 0.001). Patients who underwent CRT + S demonstrated improved overall survival compared with S alone (hazard ratio [HR] = 0.81; 95% confidence interval [CI], 0.66-0.98; p = 0.03) and RT (HR = 0.44; 95% CI, 0.35-0.55; p < 0.0001); and comparable survival to CRT (HR = 0.82; 95% CI, 0.67-1.01; p = 0.06). Patients who underwent CRT + S also had comparable postoperative mortality (HR = 0.96; 95% CI, 0.87-1.07; p = 0.45) and complications (OR = 0.89; 95% CI, 0.70-1.14; p = 0.36) compared with S alone. CONCLUSIONS: Preoperative chemoradiotherapy may be an acceptable treatment option in appropriately selected older esophageal cancer patients. This treatment modality did not appear to increase surgical complications and offered potential therapeutic benefit, particularly compared with surgery alone. PMID- 19289263 TI - Metabolic tumor volume predicts for recurrence and death in head-and-neck cancer. AB - PURPOSE: To evaluate the prognostic value of metabolic tumor volume measured on 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) imaging and other clinical factors in patients treated for locally advanced head-and-neck cancer (HNC) at a single institution. MATERIALS AND METHODS: Between March 2003 and August 2007, 85 patients received positron emission tomography (PET)/computed tomography-guided chemoradiotherapy for HNC. Metabolically active tumor regions were delineated on pretreatment PET scans semiautomatically using custom software. We evaluated the relationship of (18)F-fluorodeoxyglucose-PET maximum standardized uptake value (SUV) and total metabolic tumor volume (MTV) with disease-free survival (DFS) and overall survival (OS). RESULTS: Mean follow-up for surviving patients was 20.4 months. The estimated 2-year locoregional control, DFS, and OS for the group were 88.0%, 69.5%, and 78.4%, respectively. The median time to first failure was 9.8 months among the 16 patients with relapse. An increase in MTV of 17.4 mL (difference between the 75th and 25th percentiles) was significantly associated with an increased hazard of first event (recurrence or death) (1.9-fold, p < 0.001), even after controlling for Karnofsky performance status (KPS) (1.8-fold, p = 0.001), and of death (2.1-fold, p < 0.001). We did not find a significant relationship of maximum SUV, stage, or other clinical factors with DFS or OS. CONCLUSIONS: Metabolic tumor volume is an adverse prognostic factor for disease recurrence and death in HNC. MTV retained significance after controlling for KPS, the only other significant adverse prognostic factor found in this cohort. MTV is a direct measure of tumor burden and is a potentially valuable tool for risk stratification and guiding treatment in future studies. PMID- 19289264 TI - Phase I study of concurrent whole brain radiotherapy and erlotinib for multiple brain metastases from non-small-cell lung cancer. AB - PURPOSE: Erlotinib has shown activity in patients with brain metastases from non small-cell lung cancer. The present dose-escalation Phase I trial evaluated the toxicity of whole brain radiotherapy (WBRT) with concurrent and maintenance erlotinib in this patient group. METHODS AND MATERIALS: Erlotinib (Cohort 1, 100 mg/d; Cohort 2, 150 mg/d) was started 1 week before, and continued during, WBRT (30 Gy in 10 fractions). Maintenance erlotinib (150 mg/d) was continued until unacceptable toxicity or disease progression. RESULTS: A total of 11 patients completed WBRT, 4 in Cohort 1 and 7 in Cohort 2. The median duration of erlotinib treatment was 83 days. No treatment-related neurotoxicity was observed. No treatment-related Grade 3 or greater toxicity occurred in Cohort 1. In Cohort 2, 1 patient developed a Grade 3 acneiform rash and 1 patient had Grade 3 fatigue. Two patients in Cohort 2 developed erlotinib-related interstitial lung disease, contributing to death during maintenance therapy. The median overall survival and interval to progression was 133 and 141 days, respectively. Six patients developed extracranial progression; only 1 patient had intracranial progression. In 7 patients with follow-up neuroimaging at 3 months, 5 had a partial response and 2 had stable disease. CONCLUSION: WBRT with concurrent erlotinib is well tolerated in patients with brain metastases from non-small-cell lung cancer. The suggestion of a high intracranial disease control rate warrants additional study. PMID- 19289265 TI - Biological image-guided radiotherapy in rectal cancer: challenges and pitfalls. AB - PURPOSE: To investigate the feasibility of integrating multiple imaging modalities for image-guided radiotherapy in rectal cancer. PATIENTS AND METHODS: Magnetic resonance imaging (MRI) and fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) were performed before, during, and after preoperative chemoradiotherapy (CRT) in patients with resectable rectal cancer. The FDG-PET signals were segmented with an adaptive threshold-based and a gradient-based method. Magnetic resonance tumor volumes (TVs) were manually delineated. A nonrigid registration algorithm was applied to register the images, and mismatch analyses were carried out between MR and FDG-PET TVs and between TVs over time. Tumor volumes delineated on the images after CRT were compared with the pathologic TV. RESULTS: Forty-five FDG-PET/CT and 45 MR images were analyzed from 15 patients. The mean MRI and FDG-PET TVs showed a tendency to shrink during and after CRT. In general, MRI showed larger TVs than FDG-PET. There was an approximately 50% mismatch between the FDG-PET TV and the MRI TV at baseline and during CRT. Sixty-one percent of the FDG-PET TV and 76% of the MRI TV obtained after 10 fractions of CRT remained inside the corresponding baseline TV. On MRI, residual tumor was still suspected in all 6 patients with a pathologic complete response, whereas FDG-PET showed a metabolic complete response in 3 of them. The FDG-PET TVs delineated with the gradient-based method matched closest with pathologic findings. CONCLUSIONS: Integration of MRI and FDG-PET into radiotherapy seems feasible. Gradient-based segmentation is recommended for FDG PET. Spatial variance between MRI and FDG-PET TVs should be taken into account for target definition. PMID- 19289266 TI - Radiation dose predicts for biochemical control in intermediate-risk prostate cancer patients treated with low-dose-rate brachytherapy. AB - PURPOSE: To evaluate the influence of patient- and treatment-related factors on freedom from biochemical failure (FFbF) in patients with intermediate-risk prostate cancer. METHODS AND MATERIALS: From a prospectively collected database of 2250 men treated at Mount Sinai Hospital from 1990 to 2004 with low-dose-rate brachytherapy for prostate cancer, 558 men with either one or more intermediate risk features (prostate-specific antigen [PSA] level 10-20 ng/mL, Gleason score 7, or Stage T2b) were identified who had a minimum follow-up of 24 months and postimplant CT-based dosimetric analysis. Biologically effective dose (BED) values were calculated to compare doses from different isotopes and treatment regimens. Patients were treated with brachytherapy with or without hormone therapy and/or external-beam radiotherapy. Patient- and treatment-related factors were analyzed with respect to FFbF. The median follow-up was 60 months (range, 24 167 months). Biochemical failure was defined according to the Phoenix definition. Univariate analyses were used to determine whether any variable was predictive of FFbF. A two-sided p value of <0.05 was considered significant. RESULTS: Overall, the actuarial FFbF at 10 years was 86%. Dose (BED <150 Gy(2) vs. >or=150 Gy(2)) was the only significant predictor of FFbF (p < 0.001). None of the other variables (PSA, external-beam radiotherapy, Gleason score, treatment type, hormones, stage, and number of risk factors) was found to be a statistically significant predictor of 10-year FFbF. CONCLUSIONS: Radiation dose is an important predictor of FFbF in intermediate-risk prostate cancer. Treatment should continue to be individualized according to presenting disease characteristics until results from Radiation Therapy Oncology Group trial 0232 become available. PMID- 19289267 TI - Dose-volume histogram parameters and local tumor control in magnetic resonance image-guided cervical cancer brachytherapy. AB - PURPOSE: To investigate the value of dose-volume histogram (DVH) parameters for predicting local control in magnetic resonance (MR) image-guided brachytherapy (IGBT) for patients with cervical cancer. METHODS AND MATERIALS: Our study population consists of 141 patients with cervical cancer (Stages IB-IVA) treated with 45-50 Gy external beam radiotherapy plus four times 7 Gy IGBT with or without cisplatin. Gross tumor volume (GTV), high-risk clinical target volume (HRCTV), and intermediate-risk clinical target volume (IRCTV) were contoured, and DVH parameters (minimum dose delivered to 90% of the volume of interest [D90] and D100) were assessed. Doses were converted to the equivalent dose in 2 Gy (EQD2) by applying the linear quadratic model (alpha/beta = 10 Gy). Groups were defined for patients with or without local recurrence (LR) in the true pelvis for tumor size at diagnosis (GTV at diagnosis [GTVD] of 2-5 cm (Group 1) or greater than 5 cm (Group 2) and for tumor size response at IGBT (HRCTV) of 2-5 cm (Group 2a) or greater than 5 cm (Group 2b). RESULTS: Eighteen LRs were observed. The most important DVH parameters correlated with LR were the D90 and D100 for HRCTV. Mean D90 and D100 values for HRCTV were 86 +/- 16 and 65 +/- 10 Gy, respectively. The D90 for HRCTV greater than 87 Gy resulted in an LR incidence of 4% (3 of 68) compared with 20% (15 of 73) for D90 less than 87 Gy. The effect was most pronounced in the tumor group (Group 2b). CONCLUSIONS: We showed an increase in local control in IGBT in patients with cervical cancer with the dose delivered, which can be expressed by the D90 and D100 for HRCTV. Local control rates greater than 95% can be achieved if the D90 (EQD2) for HRCTV is 87 Gy or greater. PMID- 19289268 TI - Population-based study of trends and variations in radiotherapy as part of primary treatment of cancer in the southern Netherlands between 1988 and 2006, with an emphasis on breast and rectal cancer. AB - PURPOSE: To explore current variations in the use of primary radiotherapy (RT) in a region with two RT departments with adjacent referral areas, one in the eastern and one in the western sector of the southern region of the Netherlands. METHODS AND MATERIALS: We calculated the proportion of 147,588 patients with newly diagnosed cancer between 1988 and 2006 in the southern Netherlands who received primary RT. Especially for breast and rectal cancer patients we studied primary RT use according to stage (breast cancer) and age and separately for the eastern and western sectors. RESULTS: The number of patients with new diagnoses receiving primary RT increased from 1,668 patients in 1988 to 2,971 patients in 2006, with the proportion of the overall patients receiving RT remaining more or less unchanged (+/-30%). However, only 20% of elderly patients (75+ years) received primary RT. Over time, more patients with prostate and rectal cancer, fewer patients with lung and bladder cancer or Hodgkin's lymphoma, and, recently, more patients with cervical or endometrial cancer received RT. The proportion of patients with most other tumor types treated with RT remained more or less unchanged. The total RT rate was slightly higher for patients in the eastern sector. Of particular note, patients with breast or rectal cancer in the eastern sector were significantly more likely to receive primary RT than were their counterparts in the western sector (odds ratio = 1.4, 95% confidence interval =1.4-1.5, and odds ratio = 1.4, 95% confidence interval = 1.3-1.6, respectively). CONCLUSIONS: Although the number of RT-treated patients increased substantially during 1988 to 2006, the proportion remained essentially unchanged. In addition, large variations were found in referral rates for RT, especially in later years, between the eastern and the western sectors of the region. PMID- 19289269 TI - Guanidinoacetate methyltransferase deficiency (GAMT). AB - An increasing number of disorders of metabolism are becoming amenable to the treatment, and GAMT deficiency is one of them. The symptoms and signs are reviewed, emphasising that delayed language development is a particular feature. Other symptoms include learning disorders, autistic behaviour, epileptic seizures, and movement disorders. The condition is inherited in an autosomal recessive manner, and mutations in the GAMT gene severely affect the activity of guanidinoacetate. The MRI scan shows an increased signal in the globus pallidus, and the diagnosis is confirmed by finding increased guanidinoacetate in the urine and a low plasma creatine. Other methods of diagnosis are discussed. Treatment is based on giving creatine supplementation orally and a low-protein diet with restricted arginine and increased ornithine. This results in improvement of many of the symptoms, especially of the epileptic seizures and the abnormal movements. It is justifiable to consider this condition in any patient with unexplained learning disorders. PMID- 19289271 TI - Expected reduction of colorectal cancer incidence within 8 years after introduction of the German screening colonoscopy programme: estimates based on 1,875,708 screening colonoscopies. AB - In late 2002, colonoscopy was introduced as a primary screening tool for colorectal cancer (CRC) in Germany. We aimed to estimate the expected reduction in case numbers and incidence of CRC between 2003 and 2010 by detection and removal of advanced adenomas. Data from 1,875,708 women and men included in the national screening colonoscopy database were combined with estimates of transition rates of advanced adenomas and with national population projections. Despite relatively low screening participation, incident CRC cases are expected to be reduced by more than 15,000 between 2003 and 2010. The impact is expected to be largest in age groups 55-59, 60-64 and 65-69 in whom total case numbers in 2010 are expected to be reduced by 13%, 19% and 14% among women, and by 11%, 15% and 12%, respectively, among men. Our results forecast a major rapid reduction of the CRC burden in Germany by screening colonoscopy. PMID- 19289270 TI - Homing of GAD65 specific autoimmunity and development of insulitis requires expression of both DQ8 and human GAD65 in transgenic mice. AB - MHC-class II genes determine susceptibility in human type-1 diabetes. In their context, presentation of target antigen(s) results in autoimmunity and beta-cell destruction. An animal model, in which human beta-cell autoantigen(s) are presented to effector cells in the context of human MHC-class II diabetes susceptibility genes, would be desirable for studying molecular mechanisms of disease and developing antigen-specific immune-interventions. We report the development of antigen-specific insulitis in double-transgenic mice carrying the HLA-DQ8 diabetes-susceptibility haplotype and expressing the human autoantigen GAD65 in pancreatic beta-cells. Immunization with human GAD65 cDNA resulted in severe insulitis and low antibody levels in double-transgenic mice while control mice were mostly insulitis free. CFA/protein immunization resulted in high antibody levels and modest insulitis. Pancreatic lymphocytic infiltration progressed through stages (exocrine pancreas followed by peri- and intra insulitis). Adoptive transfer of splenocytes from DNA-immunized mice resulted in development of insulitis in recipient transgenics. Our results show that immunization with a clinically relevant, type-1 diabetes human autoantigen, in a humanized genetic setting, results in the development of an immune response that homes to islets of Langerhans. This animal model will facilitate studies of autoimmunity to GAD65 in the context of HLA-DQ8, and development of methods to induce tolerance and prevent insulitis. PMID- 19289272 TI - First air-tolerant effective stainless steel microbial anode obtained from a natural marine biofilm. AB - Microbial anodes were constructed with stainless steel electrodes under constant polarisation. The seawater medium was inoculated with a natural biofilm scraped from harbour equipment. This procedure led to efficient microbial anodes providing up to 4A/m(2) for 10mM acetate oxidation at -0.1 V/SCE. The whole current was due to the presence of biofilm on the electrode surface, without any significant involvement of the abiotic oxidation of sulphide or soluble metabolites. Using a natural biofilm as inoculum ensured almost optimal performance of the biofilm anode as soon as it was set up; the procedure also proved able to form biofilms in fully aerated media, which provided up to 0.7A/m(2). The current density was finally raised to 8.2A per square meter projected surface area using a stainless steel grid. The inoculating procedure used here combined with the control of the potential revealed, for the first time, stainless steel as a very competitive material for forming bioanodes with natural microbial consortia. PMID- 19289273 TI - Simultaneous saccharification and fermentation of lignocellulosic residues pretreated with phosphoric acid-acetone for bioethanol production. AB - Bermudagrass, reed and rapeseed were pretreated with phosphoric acid-acetone and used for ethanol production by means of simultaneous saccharification and fermentation (SSF) with a batch and fed-batch mode. When the batch SSF experiments were conducted in a 3% low effective cellulose, about 16 g/L of ethanol were obtained after 96 h of fermentation. When batch SSF experiments were conducted with a higher cellulose content (10% effective cellulose for reed and bermudagrass and 5% for rapeseed), higher ethanol concentrations and yields (of more than 93%) were obtained. The fed-batch SSF strategy was adopted to increase the ethanol concentration further. When a higher water-insoluble solid (up to 36%) was applied, the ethanol concentration reached 56 g/L of an inhibitory concentration of the yeast strain used in this study at 38 degrees C. The results show that the pretreated materials can be used as good feedstocks for bioethanol production, and that the phosphoric acid-acetone pretreatment can effectively yield a higher ethanol concentration. PMID- 19289274 TI - Application of aerobic microorganisms in bioremediation in situ of soil contaminated by petroleum products. AB - Aerobic microorganisms able to biodegrade benzene, toluene, ethylbenzene, xylene (BTEX) have been isolated from an area contaminated by petroleum products. The activity of the isolated communities was tested under both laboratory and field conditions. Benzene, toluene, ethylbenzene and xylene were added to the cultures as the sole carbon source, at a concentration of 500 mg/L. In batch cultures under laboratory conditions, an 84% reduction of benzene, 86% of toluene and 82% of xylene were achieved. In cultures with ethylbenzene as the sole carbon source, the reduction was around 80%. Slightly lower values were observed under field conditions: 95% reduction of benzene and toluene, 81% of ethylbenzene and 80% of xylene. A high biodegradation activity of benzene (914 microM/L/24h), toluene (771 microM/L/24h), xylene (673 microM/L/24h) and ethylbenzene (644 microM/L/24h) was observed in the isolated communities. PMID- 19289275 TI - Bioenergy from Coastal bermudagrass receiving subsurface drip irrigation with advance-treated swine wastewater. AB - Coastal bermudagrass (Cynodon dactylon L.) may be a potentially important source of bio-based energy in the southern US due to its vast acreage. It is often produced as part of a waste management plan with varying nutrient composition and energy characteristics on fields irrigated with livestock wastewater. The objective of this study was to determine the effect of subsurface drip irrigation with treated swine wastewater on both the quantity and quality of bermudagrass bioenergy. The treated wastewater was recycled from an advanced treatment system and used for irrigation of bermudagrass in two crop seasons. The experiment had nine water and drip line spacing treatments arrayed in a randomized complete block-design with four replicates. The bermudagrass was analyzed for calorific and mineral contents. Bermudagrass energy yields for 2004 and 2005 ranged from 127.4 to 251.4MJ ha(-1). Compared to irrigation with commercial nitrogen fertilizer, the least biomass energy density was associated with bermudagrass receiving treated swine wastewater. Yet, in 2004 the wastewater irrigated bermudagrass had greater hay yields leading to greater energy yield per ha. This decrease in energy density of wastewater irrigated bermudagrass was associated with increased concentrations of K, Ca, and Na. After thermal conversion, these compounds are known to remain in the ash portion thereby decreasing the energy density. Nonetheless, the loss of energy density using treated effluent via SDI may be offset by the positive influence of these three elements for their catalytic properties in downstream thermal conversion processes such as promoting a lesser char yield and greater combustible gas formation. PMID- 19289276 TI - Survey of US fuel ethanol plants. AB - The ethanol industry is growing in response to increased consumer demands for fuel as well as the renewable fuel standard. Corn ethanol processing creates the following products: 1/3 ethanol, 1/3 distillers grains, and 1/3 carbon dioxide. As the production of ethanol increases so does the generation of its coproducts, and viable uses continually need to be developed. A survey was mailed to operational US ethanol plants to determine current practices. It inquired about processes, equipment used, end products, and desired future directions for coproducts. Results indicated that approximately one-third of plant managers surveyed expressed a willingness to alter current drying time and temperature if it could result in a higher quality coproduct. Other managers indicated hesitation, based on lack of economic incentives, potential cost and return, and capital required. Respondents also reported the desire to use their coproducts in some of the following products: fuels, extrusion, pellets, plastics, and human food applications. These results provide a snapshot of the industry, and indicate that operational changes to the current production of DDGS must be based upon the potential for positive economic returns. PMID- 19289277 TI - Treatment of industrial wastewater with two-stage constructed wetlands planted with Typha latifolia and Phragmites australis. AB - Industrial wastewater treatment comprises several processes to fulfill the discharge permits or to enable the reuse of wastewater. For tannery wastewater, constructed wetlands (CWs) may be an interesting treatment option. Two-stage series of horizontal subsurface flow CWs with Phragmites australis (UP series) and Typha latifolia (UT series) provided high removal of organics from tannery wastewater, up to 88% of biochemical oxygen demand (BOD(5)) (from an inlet of 420 to 1000 mg L(-1)) and 92% of chemical oxygen demand (COD) (from an inlet of 808 to 2449 mg L(-1)), and of other contaminants, such as nitrogen, operating at hydraulic retention times of 2, 5 and 7 days. No significant (P<0.05) differences in performance were found between both the series. Overall mass removals of up to 1294 kg COD ha(-1)d(-1) and 529 kg BOD(5)ha(-1)d(-1) were achieved for a loading ranging from 242 to 1925 kg COD ha(-1)d(-1) and from 126 to 900 kg BOD(5)ha(-1)d( 1). Plants were resilient to the conditions imposed, however P. australis exceeded T. latifolia in terms of propagation. PMID- 19289278 TI - Kinetics of an extracellular exo-inulinase production from a 5-flourocytosine resistant mutant of Geotrichum candidum using two-factorial design. AB - In the present study, eight different strains of Geotrichum candidum were isolated and screened for an extracellular exo-inulinase production using chemically enriched sucrose-mineral media. The isolate (Zool-3i) with a better enzyme activity (1.38 IU/ml) was subjected to induced mutagenesis using methyl methane sulphonate (MMS) and a mutant with an enzyme activity of 32.06 IU/ml was obtained. Further exposure to ethyl methane sulphonate (EMS) and ultraviolet (UV) radiations yielded a mutant exhibiting an improved activity of 39.34 IU/ml. The potential mutant was cultured overnight and plated on 5fc-YPR agar medium and thus made resistant against 5-flourocytocine. Over 50-fold enhancement in enzyme production (71.85 IU/ml) was achieved when the process parameters including incubation period (48 h), sucrose concentration (5.0 g/L), pH (6.0), inoculum size (2.0%, 16 h old) and urea (0.2%) were identified using Plackett-Burman design. On the basis of kinetic variables, notably Q(p) (0.723 U/g/h), Y(p/s) (2.036U/g) and q(p) (0.091U/g cells/h), the mutant MEU-5fc-6 was found to be a hyper producer of exo-inulinase (HS, LSD 0.045, p< or =0.05). PMID- 19289279 TI - Restricted spacer tolerance of a zinc finger nuclease with a six amino acid linker. AB - Zinc finger nucleases can be engineered to create highly efficient and precise changes to the genetic information within living cells. We report the investigation of an important parameter that defines the type of target site the nuclease can cleave. The active nuclease is a dimer, requiring that the DNA target site contain two zinc finger binding sites separated by a short spacer. Using a plasmid-based recombination assay in HEK 293T cells, we show that a 6 amino acid linker between the zinc finger DNA-binding domain and the FokI cleavage domain restricts nuclease activity to sites containing a 6 bp spacer. These observations concur with other recent studies, suggesting this information will be useful in the design of new potent and accurate zinc finger nucleases. PMID- 19289280 TI - [35S]GTP gamma S binding studies of amphiphilic drugs-activated Gi proteins: a caveat. AB - This paper documents a serious problem met during the testing of Gi protein activating properties of a new series of synthetic compounds by measuring the induced binding of [(35)S]GTPgammaS to different subtypes of Gi protein. The problem arose from the strong affinity between [(35)S]GTPgammaS and the tested compounds, that are characterized by several (2-4) positive charges and high lipophilicity. Apparently, such affinity yields insoluble, labelled complexes that, also in the absence of Gi protein, are retained on the filters and give rise to false positive results. PMID- 19289281 TI - Anti-HBV agents. Part 2: synthesis and in vitro anti-hepatitis B virus activities of alisol A derivatives. AB - Chemical modifications were performed on hydroxyl groups at C-11,23,24,25 positions and C-13(17) double bond of alisol A for structure-activity relationship study. Forty-one derivatives of alisol A were synthesized and assayed for their in vitro anti-hepatitis B virus (HBV) activities and cytotoxicities. Of them, 14 compounds were active against HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) secretion in HepG 2.2.15 cells, and the most promising compound 25 exhibited high activities against secretion of HBsAg (IC(50)=0.028 mM), HBeAg (IC(50)=0.027 mM) and remarkable selective indices (SI(HBsAg) >90, SI(HBeAg) >93). PMID- 19289282 TI - Ganoderic acid DM: anti-androgenic osteoclastogenesis inhibitor. AB - Prostate cancer is the most common cancer in men in Western countries, with a high incidence of bone metastasis. Ganoderic acid DM, with 5alpha-reductase inhibitory and androgen receptor (AR) binding activity, isolated from the ethanol extracts of Ganoderma lucidum, can inhibit prostate cancer cell growth and block osteoclastogenesis. PMID- 19289283 TI - In vitro and in vivo SAR of pyrido[3,4-d]pyramid-4-ylamine based mGluR1 antagonists. AB - The SAR of a series of novel pyrido[3,4-d]pyramid-4-ylamine mGluR1 antagonists is described. The multiple of the unbound K(i) in cerebrospinal fluid necessary to give morphine like analgesic effects in an electromyograph pinch model in rodents is determined and the effect of structure on CNS penetration examined. PMID- 19289284 TI - Pectoralis major interposition flap: a new technique for treatment of severe peri ductal mastitis. AB - BACKGROUND: Peri-ductal mastitis is an uncommon benign disorder of the major lactiferous ductal system of the female breast. It can be a very difficult problem to treat and may cause significant patient morbidity. We describe a new technique, involving use of the pectoralis major muscle flap, for treating recurrent sub-areolar abscess refractory to standard surgical treatment. METHOD: Three patients who underwent this new technique for severe refractory peri-ductal mastitis at Calvary Hospital, Canberra are presented. RESULTS: These patients who had recurrent peri-ductal mastitis with abscess and fistula formation on a monthly basis despite numerous courses of antibiotics and surgical procedures experienced no further recurrences following pectoralis major interposition flap surgery at 42, 32 and 22 months follow-up respectively. CONCLUSION: This new technique may provide an opportunity to control these otherwise difficult to treat cases of severe recurrent peri-ductal mastitis where standard surgical methods have failed. PMID- 19289285 TI - Volume-controlled vs no/short-term drainage after axillary lymph node dissection in breast cancer surgery: a meta-analysis. AB - It is unknown whether there are any clinically relevant differences between volume-controlled (<30-50 ml/24h across trials) vs no/short-term drainage after axillary lymph node dissection in breast cancer surgery on outcomes such as seroma formation, wound infection or length of hospital stay. Randomised controlled trials comparing volume-controlled drainage vs no or short-term drainage after axillary lymph node dissection in breast cancer surgery were identified systematically using Pubmed, EMBASE and The Cochrane library. Trial data were reviewed and extracted independently by two reviewers in a standardised unblinded manner. Six randomised controlled trials which included a total of 561 patients fulfilled our inclusion criteria. Patients randomised to volume controlled drainage were less likely to develop clinically relevant seromas compared to patients randomised to no/short-term drainage. There was, however, no difference in wound infections between patients treated with volume-controlled drainage and patients with no or short-term drainage. Patients randomised to volume-controlled drainage stayed significantly longer in hospital than patients randomised to no/short-term drainage. Based on available evidence, clinically relevant seromas occur more frequently in patients treated with no/short-term drainage. However, no/short-term drainage after axillary lymph node dissection does not lead to an increase in wound infections and is associated with shorter hospital stay. PMID- 19289286 TI - Recombinant ciliary neurotrophic factor promotes nerve regeneration and induces gene expression in silicon tube-bridged transected sciatic nerves in adult rats. AB - Sciatic nerves in adult male rats were transected and reunited via a silicone chamber. This was followed by a focal injection of recombinant ciliary neurotrophic factor (CNTF). To evaluate the effect of this therapeutic approach and to explore its possible mechanisms, nerve regeneration was traced by horseradish peroxidase retrograde labeling. Functional recovery was evaluated by functional assessment of the hind feet and the expression of a number of proteins was detected using immunohistochemistry. The results showed that a single administration of CNTF could promote regeneration of motor axons, with improved functional recovery in adult rats. Growth associated protein (GAP)-43, S100, CD68 and major histocompatibility complex class II immunoreactivity in the regenerative and distal nerves suggested that CNTF could promote axon regeneration, Schwann cell migration, monocyte infiltration and activation. CNTF might also indirectly promote axonal regeneration by further activating the JAK STAT3 pathway and subsequently upregulating phosphotyrosine, GAP-43 and S100 expression to enhance proliferation, growth and migration of Schwann cells. CNTF has suggested important targets for pharmacological intervention in peripheral nerve disease and injury. PMID- 19289287 TI - Surgical complications after endoscopic transsphenoidal pituitary surgery. AB - Between January 2004 and June 2007 we conducted a retrospective analysis to assess post-operative complications related to endoscopic pituitary surgery in a series of 150 patients. Patients were treated with an endoscopic endonasal transsphenoidal approach to the sellar region for removal of pathological sellar and suprasellar lesions. We analysed the complications in groups according to the anatomical structures of the approach and the functional systems of the pituitary gland (anterior and posterior endocrine systems), and compared them to a large historical series using the traditional microsurgical transsphenoidal approach. Overall, we observed a decreased incidence of complications with respect to the surgical trauma, the functionality of the pituitary gland and post-operative patient comfort. We believe that the reduction of the complication rate observed in this study was mainly due to the wide structural overview offered by the endoscope as well as the anatomically direct, and therefore minimally invasive, character of the procedure. Successful endoscopic pituitary surgery requires extensive training in the use of an endoscope and careful planning of the surgery. Furthermore, close cooperation between a multidisciplinary team consisting of endocrinologists, neurosurgeons, ear, nose and throat surgeons, radiologists, and radiation oncologists is of utmost importance. PMID- 19289288 TI - Fenestration of the middle cerebral artery with an associated ruptured aneurysm. AB - Intracranial arterial fenestrations are uncommon developmental anomalies that may alter local hemodynamic stress patterns and may be associated with intracranial aneurysms. A 75-year-old woman presented with a mild subarachnoid hemorrhage. Cerebral angiography demonstrated a 5 mm aneurysm arising from the middle cerebral artery (MCA) associated with a dramatic fenestration of the proximal M1 segment of the MCA. The patient underwent craniotomy with microsurgical aneurysm clipping. The aneurysm arose immediately distal to the fenestration. Several fine perforating arteries were dissected away from the neck of the aneurysm to allow for safe clip placement. Postoperatively, the patient awoke without deficit and was discharged to home on the fifth postoperative day in good condition. Anomalies of the intracranial vasculature are common and we describe a rare proximal MCA fenestration with an associated ruptured aneurysm at the site of the fenestration. Neurosurgeons should be thoroughly familiar with such potential "normal variants" to avoid operative complications that may result from the anomalous circulation in such cases. PMID- 19289289 TI - A case report comparing 3-D evaluation in the diagnosis and treatment planning of hemimandibular hyperplasia with conventional radiography. AB - INTRODUCTION: Cephalometric radiography is the main investigation used in the diagnosis of maxillofacial deformities. Three-dimensional (3-D) computed tomography (3-D CT) imaging has recently been used in the diagnosis and treatment planning of craniofacial morphology. The purpose of this report was to present the diagnosis and treatment planning of a hemimandibular hyperplasia case by using conventional radiographs and 3-D cephalometric analysis. METHODS: A 24-year old girl presented with hemimandibular hyperplasia and the main complaint was her facial appearance. Conventional cephalometric analysis including lateral and frontal cephalograms were traced by hand and evaluated by the same clinician. Twenty-one measurements on the lateral cephalograms and 13 measurements on the frontal cephalograms were performed. In addition, axial images were reconstructed into a 3-D model by using medical image processing software. Linear and angular measurements were presented in coronal and sagittal planes. RESULTS: The 3-D CT analysis has a superior image quality when compared with conventional cephalograms. CONCLUSIONS: The 3-D CT analysis provides more advanced diagnostic image information in evaluating craniofacial morphology. We conclude that 3-D CT images can facilitate the diagnosis, treatment planning, and outcome evaluation of severe asymmetry cases. PMID- 19289290 TI - Plasma fractionation issues. AB - Procurement and processing of human plasma for fractionation of therapeutic proteins or biological medicines used in clinical practice is a multi-billion dollar international trade. Together the private sector and public sector (non profit) provide large amounts of safe and effective therapeutic plasma proteins needed worldwide. The principal therapeutic proteins produced by the dichotomous industry include gamma globulins or immunoglobulins (including pathogen-specific hyperimmune globulins, such as hepatitis B immune globulins) albumin, factor VIII and Factor IX concentrates. Viral inactivation, principally by solvent detergent and other processes, has proven highly effective in preventing transmission of enveloped viruses, viz. HBV, HIV, and HCV. PMID- 19289291 TI - Perceiving one's own movements when using a tool. AB - The present study examined what participants perceive of their hand movements when using a tool. In the experiments different gains for either the x-axis or the y-axis perturbed the relation between hand movements on a digitizer tablet and cursor movements on a display. As a consequence of the perturbation participants drew circles on the display while their covered hand movements followed either vertical or horizontal ellipses on the digitizer tablet. When asked to evaluate their hand movements, participants were extremely uncertain about their trajectories. By varying the amount of visual feedback, findings indicated that the low awareness of one's own movements originated mainly from an insufficient quality of the humans' tactile and proprioceptive system or from an insufficient spatial reconstruction of this information in memory. PMID- 19289292 TI - Suggested visual hallucinations in and out of hypnosis. AB - We administered suggestions to see a gray-scale pattern as colored and a colored pattern in shades of gray to 30 high suggestible and eight low suggestible students. The suggestions were administered twice, once following the induction of hypnosis and once without an induction. Besides rating the degree of color they saw in the stimuli differently, participants also rated their states of consciousness as normal, relaxed, hypnotized, or deeply hypnotized. Reports of being hypnotized were limited to highly suggestible participants and only after the hypnotic induction had been administered. Reports of altered color perception were also limited to high suggestibles, but were roughly comparable regardless of whether hypnosis had been induced. These data indicate that suggestible individuals do not slip into a hypnotic state when given imaginative suggestions without the induction of hypnosis, but nevertheless report experiencing difficult suggestions for profound perceptual alterations that are pheonomenologically similar to what they report in hypnosis. PMID- 19289293 TI - Simultaneous two-voxel localized (1)H-observed (13)C-edited spectroscopy for in vivo MRS on rat brain at 9.4T: Application to the investigation of excitotoxic lesions. AB - (13)C spectroscopy combined with the injection of (13)C-labeled substrates is a powerful method for the study of brain metabolism in vivo. Since highly localized measurements are required in a heterogeneous organ such as the brain, it is of interest to augment the sensitivity of (13)C spectroscopy by proton acquisition. Furthermore, as focal cerebral lesions are often encountered in animal models of disorders in which the two brain hemispheres are compared, we wished to develop a bi-voxel localized sequence for the simultaneous bilateral investigation of rat brain metabolism, with no need for external additional references. Two sequences were developed at 9.4T: a bi-voxel (1)H-((13)C) STEAM-POCE (Proton Observed Carbon Edited) sequence and a bi-voxel (1)H-((13)C) PRESS-POCE adiabatically decoupled sequence with Hadamard encoding. Hadamard encoding allows both voxels to be recorded simultaneously, with the same acquisition time as that required for a single voxel. The method was validated in a biological investigation into the neuronal damage and the effect on the Tri Carboxylic Acid cycle in localized excitotoxic lesions. Following an excitotoxic quinolinate-induced localized lesion in the rat cortex and the infusion of U-(13)C glucose, two (1)H-((13)C) spectra of distinct (4x4x4mm(3)) voxels, one centred on the injured hemisphere and the other on the contralateral hemisphere, were recorded simultaneously. Two (1)H bi-voxel spectra were also recorded and showed a significant decrease in N acetyl aspartate, and an accumulation of lactate in the ipsilateral hemisphere. The (1)H-((13)C) spectra could be recorded dynamically as a function of time, and showed a fall in the glutamate/glutamine ratio and the presence of a stable glutamine pool, with a permanent increase of lactate in the ipsilateral hemisphere. This bi-voxel (1)H-((13)C) method can be used to investigate simultaneously both brain hemispheres, and to perform dynamic studies. We report here the neuronal damage and the effect on the Tri Carboxylic Acid cycle in localized excitotoxic lesions. PMID- 19289294 TI - Dysphagia and hypervitaminosis A: cervical hyperostosis. AB - Vertebral hyperostosis typically predominates at the thoracic spine and causes only minor symptoms. Involvement of the cervical spine may cause dysphagia due to pressure on the esophagus. We report three cases of dysphagia revealing cervical hyperostosis. CASE REPORTS: The patients were 3 men aged 54-73 years. Dysphagia was moderate in 2 patients and severe in 1 patient who had lost 4 kg over 6 months. Stiffness of the neck with mild pain was present. One patient reported a neck injury in childhood and another had a brother and father with similar symptoms. Radiographs showed exuberant anterior cervical hyperostosis. Two patients also had hyperostotic changes at the thoracic spine and pelvis. The skin and neurological evaluation were normal. Findings were normal from standard blood tests (C-reactive protein, calcium, and vitamin D). Tests were negative for the HLA-B27 antigen in all 3 patients. Serum vitamin A levels were high, ranging from 894 to 1123 microg/L (normal, 489-720). None of the patients reported taking retinoids or having unusual eating habits. DISCUSSION: Dysphagia can result from anterior cervical hyperostosis. A role for hypervitaminosis A in the genesis of hyperostosis has long been suspected. In our patients, the absence of vitamin A supplementation suggests an abnormality in vitamin A metabolism. PMID- 19289295 TI - Anakinra is a possible alternative in the treatment and prevention of acute attacks of pseudogout in end-stage renal failure. AB - We describe the case of a 71-year-old man with recurrent pseudogout attacks affecting multiple joints. He had end-stage renal failure that contra-indicated the use of non-steroidal anti-inflammatory drugs and was resistant to therapy with glucocorticoids. Based on the recent findings that interleukin (IL)-1beta is involved in crystal-induced inflammation, the patient received anakinra, a specific IL-1 inhibitor, in order to treat an acute attack of pseudogout. In addition, anakinra was administered as preventive therapy 3days per week after each hemodialysis session. Under this treatment, he did not present any severe episode of arthritis after a follow-up of 8 months. This observation suggests that anakinra is efficacious and safe for the prevention of crystal-induced arthritis in patients with severe renal failure. PMID- 19289296 TI - Adalimumab-induced neutropenia in a patient with rheumatoid arthritis. PMID- 19289297 TI - What can we learn from the presence of anti-cyclic citrullinated peptide antibodies in systemic lupus erythematosus? AB - BACKGROUND: Anti-cyclic citrullinated peptide (anti-CCP) antibodies have proved to be a specific marker for the diagnosis of rheumatoid arthritis (RA). However, the antibodies can also be detected in other rheumatic diseases, especially systemic lupus erythematosus (SLE). Recent studies have shown anti-CCP antibodies are associated with erosive arthritis in SLE patients. Since erosive arthritis is not common in SLE and many patients with non-erosive arthritis also have anti-CCP antibodies, the clinical significance of anti-CCP antibodies in SLE needs to be further studied. OBJECTIVE: To investigate the prevalence and clinical significance of anti-CCP antibodies in Chinese SLE patients. METHODS: Serum samples from 138 SLE patients were examined for anti-CCP with the second generation anti-CCP detection kit. The associations of anti-CCP with clinical and laboratory features, especially arthritis, in such SLE patients were analyzed. RESULTS: The prevalence of anti-CCP was 13.8% (19/138) in Chinese SLE patients. Seventy of 138 SLE patients had experienced arthritis, of whom 14 patients were anti-CCP+. Significantly, anti-CCP antibodies were more frequently found in SLE patients with arthritis than without arthritis (20% vs 7.4%, P<0.05). A statistical correlation between anti-CCP and rheumatoid factor (RF) was found in SLE patients with arthritis (r=0.36, P=0.002). The frequency of arthritis was significantly higher in SLE patients with anti-CCP than without (73.7% vs 47.1%,P<0.05). Eight out of 138 SLE patients showed joint erosions on radiographs. When compared with anti-CCP- patients, erosive arthritis occurred more often in anti-CCP+ patients (35.7% vs 5.4%, P<0.001). Interestingly, two patients without anti-CCP and RF who had erosive arthritis were anti-RA33 antibodies positive. All of 8 SLE patients with erosive arthritis in our study fulfilled 1987 ACR criteria for RA. With regard to other clinical and laboratory features, there were no differences between SLE patients with arthritis and without or between anti-CCP+ patients and anti-CCP- patients. CONCLUSIONS: Anti CCP antibodies have a frequency of 13.8% in Chinese SLE patients and its presence is closely associated with the onset of arthritis and bone erosion. PMID- 19289298 TI - Atypical forms of syphilis: two cases. AB - Syphilis is a sexually transmitted disease caused by the spirochete Treponema pallidum. A chancre usually develops initially. Organ involvement and neurological complications may occur, sometimes several years after the initial exposure. We managed two patients with syphilis responsible for joint or neurological manifestations, diagnosed in 2008. One patient presented with oligoarthritis involving the knees and right elbow, coinciding with a maculopapular and pustular eruption. In the other patient, meningoradiculitis involving the T8, T9, and T10 metameres prompted a test for Lyme disease, which was weakly positive, leading to evaluation for false-positivity due to a cross reaction. Neither patient was infected with the HIV. PMID- 19289299 TI - Identification using DNA from skin contact: case reports. AB - The DNA content of different types of forensic materials can vary substantially. "Trace DNA" is the minute quantity of DNA transferred through skin contact. Here, we report three cases of identification of trace DNA using conventional short tandem repeat (STR) or mitochondrial DNA (mtDNA). DNA was successfully obtained from fabrics by swabbing or by direct extraction and subjected to STR genotyping or mtDNA typing. In two cases, there was no amplification of PCR products containing the STR loci. This indicates that the areas chosen for DNA extraction contained trace DNA and DNA from more than one source. Therefore, it is important for forensic investigators performing DNA typing to know where an item has been frequently touched by victims and/or offenders as this will influence the choice of sites on the item to be used for DNA extraction. PMID- 19289300 TI - Identification of animal species using the partial sequences in the mitochondrial 16S rRNA gene. AB - This study investigated a PCR direct sequencing method for species identification by analyzing partial sequences in the mitochondrial 16S rRNA genes of many animal species amplified with universal primers. Samples from 182 vertebrates and 103 invertebrates were analyzed, and the sequences could be obtained in 182 and 72 species, respectively. The sequence divergence was sufficient to identify the species at the level of genus with the aid of the GenBank database and the BLAST tool. This method could be a powerful tool for animal species identification, especially in forensic cases in which many unknown biological samples should be analyzed. PMID- 19289301 TI - SNP association and sequence analysis of the NOS1AP gene in SIDS. AB - One of the speculated causes for sudden infant death syndrome (SIDS) is hereditary disease, in which long QT in electrocardiogram has been investigated in the view of mutations in various ion channel genes. In the present study, a novel QT interval determinant of SNP (rs10494366) in NOS1AP is genotyped in SIDS subjects (n=42) and the control group (n=210). Subjects carrying TT genotype was significantly associated with SIDS, compared with those carrying the TG of GG genotype (95% confidence interval 1.28-8.45). Sequence analysis revealed that one non-synonymous substitution in exon 8 (rs12817159) was observed in one subject, in addition to six common SNPs in exons and introns. This postmortem association study showed variations in NOS1AP might be involved in occurrence of SIDS. PMID- 19289302 TI - Application of SNPs in forensic casework: identification of pathological and autoptical specimens due to sample mix-up. AB - The analysis of single nucleotide polymorphisms (SNPs) together with conventional short tandem repeat (STR) and mitochondrial DNA (mtDNA) typing provide a forensic genetic approach for the identification of pathological and autoptical specimens in cases where the average length of DNA fragments is shorter than 150 bp in highly degraded samples. We applied a forensic genetic approach to digesta accidentally left after a training autopsy. PCR products were not amplified from samples containing the STR loci or common sequences used for mtDNA typing. The application of SNPs and deletion polymorphisms provides an alternative approach for DNA typing analysis. PMID- 19289303 TI - General comments and discussion: Social issues of suicide. PMID- 19289304 TI - Real-time PCR method for identification of Asian populations in forensic casework. AB - The priorities for mass disaster victim identification are rapid investigation turn-around time and low cost. We describe a DNA typing procedure to selectively identify members of Asian populations by a real-time PCR method using polymorphisms of the alpha2 chain of the type I collagen gene (COL1A2) and mitochondrial DNA (mtDNA). Among the 50 members of the Asian population included in the present study, 37 harbored a deleted allele in intron 33 of COL1A2 (26822 26823del) or the 10400C>T substitution mutation in mtDNA to give a probability of 0.740 for these SNPs in the Asian population. PMID- 19289305 TI - Medico-legal autopsies as a source of information to improve patient safety. AB - Clear data on the epidemiology of medical malpractice are missing, at least for Germany. Especially data on the frequency of malpractice claims in penal and civil law are not available. However, from epidemiological and healthcare research some data on the frequency of adverse events (AE), preventable adverse events (PAE) and negligent adverse events (NAE) are available. According to a review of the German Alliance of Patient Safety adverse events can be expected in 5-10%, preventable adverse events in 2-4%, negligent adverse events in about 1% and lethal outcome in about 0.1% of hospitalized patients. However, these data are not compatible with figures from civil and penal law and there seems to be a great "litigation gap". Data on malpractice cases are available from the files of the arbitration committees on medical malpractice, files of liability insurance companies and files of the institutes of forensic medicine. However, these files are not complete and data sources may have some interferences. The arbitration committees are, however, mainly dealing with living patients. Lethal cases are a special subgroup and the best available data source are the files of the institutes of forensic medicine. This subgroup is of special importance since death is the most severe outcome of medical malpractice and the reproach to have caused the death of a patient by medical negligence is the most severe malpractice claim. From a retrospective multicentre study on autopsies performed in cases of medical malpractice claims (altogether 101.358 autopsies, 4450 due to medical malpractice claims) the most important data will be presented (disciplines concerned, cause of accusation, classification of accusation, causes of preliminary proceedings, occupational group and number of approved medical malpractice, outcome of medical malpractice proceedings). However, these data are not only of a descriptive value but have utmost importance also for risk analysis and to increase patient safety. A thorough evaluation of serious incidences, although less sophisticated than a root cause analysis, produces far more information than the usual hospital reporting system. Identification and evaluation of errors as well as reporting of errors may also contribute to the prevention of errors which is among other medical disciplines also a task of forensic medicine. PMID- 19289306 TI - Unconfined compression properties of a porous poly(vinyl alcohol)-chitosan-based hydrogel after hydration. AB - A poly(vinyl alcohol) (PVA) hydrogel composite scaffold containing N,O carboxymethylated chitosan (NOCC) was tested to assess its potential as a scaffold for cartilage tissue engineering in a weight-bearing environment. The mechanical properties under unconfined compression for different hydration periods were investigated. The effect of supplementing PVA with NOCC (20wt.% PVA:5vol.% NOCC) produced a porosity of 43.3% and this was compared against a non porous PVA hydrogel (20g PVA: 100ml of water, control). Under non-hydrated conditions, the porous PVA-NOCC hydrogel behaved in a similar way to the control non-porous PVA hydrogel, with similar non-linear stress-strain response under unconfined compression (0-30% strain). After 7days' hydration, the porous hydrogel demonstrated a reduced stiffness (0.002kPa, at 25% strain), resulting in a more linear stiffness relationship over a range of 0-30% strain. Poisson's ratio for the hydrated non-porous and porous hydrogels ranged between 0.73 and 1.18, and 0.76 and 1.33, respectively, suggesting a greater fluid flow when loaded. The stress relaxation function for the porous hydrogel was affected by the hydration period (from 0 to 600s); however the percentage stress relaxation regained by about 95%, after 1200s for all hydration periods assessed. No significant differences were found between the different hydration periods between the porous hydrogels and control. The calculated aggregate modulus, H(A), for the porous hydrogel reduced drastically from 10.99kPa in its non-hydrated state to about 0.001kPa after 7days' hydration, with the calculated shear modulus reducing from 30.92 to 0.14kPa, respectively. The porous PVA-NOCC hydrogel conformed to a biphasic, viscoelastic model, which has the desired properties required for any scaffold in cartilage tissue engineering. PMID- 19289307 TI - An electrochemical study on self-ordered nanoporous and nanotubular oxide on Ti 35Nb-5Ta-7Zr alloy for biomedical applications. AB - Highly ordered nanoporous and nanotubular oxide layers were developed on low rigidity beta Ti-35Nb-5Ta-7Zr alloy by controlled DC anodization in electrolyte containing 1M H(3)PO(4) and 0.5wt.% NaF at room temperature. The as-formed and crystallized nanotubes were characterized by electron microscopy, energy dispersive X-ray spectrometry and X-ray diffraction. The electrochemical passivation behavior of the nanoporous and nanotubular oxide surfaces were investigated in Ringer's solution at 37+/-1 degrees C employing a potentiodynamic polarization technique and impedance spectroscopy. The diameters of the as-formed nanotubes were in the range of 30-80nm. The nanotubular surface exhibited passivation behavior similar to that of the nanoporous surface. However, the corrosion current density was considerably higher for the nanotubular alloy. The surface after nanotube formation seemed to favor an immediate and effective passivation. Electrochemical impedance spectra were simulated by equivalent circuits and the results were discussed with regard to biomedical applications. PMID- 19289308 TI - Synthesis and characterization of a novel N-vinylcaprolactam-containing acrylic acid terpolymer for applications in glass-ionomer dental cements. AB - In this study a novel N-vinylcaprolactam (NVC)-containing copolymer of acrylic itaconic acid was synthesized, characterized and incorporated into Fuji IX conventional glass-ionomer cement (GIC). Subsequently, the effects of incorporation of synthesized terpolymer on the mechanical properties of GIC were studied. The synthesized terpolymer was characterized using (1)H nuclear magnetic resonance, Fourier transform infrared and Raman spectroscopy. The viscosity and molecular weight of the terpolymer were also measured. The compressive strength (CS), diametral tensile strength (DTS) and biaxial flexural strength (BFS) of the modified GICs were evaluated after 24h and 1week of immersion in distilled water at 37 degrees C. The handling properties (working and setting times) of the resulting modified cements were also evaluated. One-way analysis of variance was used to study the statistical significance of the mechanical strengths and handling properties in comparison to the control group. The results showed that NVC-containing GIC samples exhibited significantly higher (P<0.05) DTS (38.3+/ 10.9MPa) and BFS (82.2+/-12.8MPa) in comparison to Fuji IX GIC (DTS=19.6+/ 11.4MPa; BFS=41.3+/-10.5MPa). The experimental cement also showed higher but not statistically significant values for CS compared to the control material (CS for NVC-containing sample=303+/-32.8MPa; CS for Fuji XI=236+/-41.5MPa). Novel NVC containing GIC has been developed in this study, with a 28% increase in CS. The presented GIC is capable of doubling the DTS and BFS in comparison to commercial Fuji IX GIC. The working properties of NVC-containing glass-ionomer formulations are comparable and are acceptable for water-based cements. PMID- 19289309 TI - Baseline Quality of Life in patients with Klippel-Trenaunay syndrome. AB - BACKGROUND: Klippel-Trenaunay syndrome (KTS) is a congenital group of disorders characterised by vascular malformations (capillary malformation (CM), venous malformation (VM), and lymphatic malformation (LM)) and disturbed growth regulation. The burden caused by KTS symptoms can be evaluated using Quality of Life (QoL)-measuring questionnaires. This study aimed to assess the QoL in KTS patients using the Short Form Health Survey Questionnaire (SF-36) and Skindex-29 questionnaires, and to determine three grades of severity (mild, moderate and severe) according to the scores obtained. In addition, we compared the SF-36 results to those of a general Dutch population sample and a selected group of other chronic conditions. METHODS: KTS patients of the Dutch KTS foundation and of two medical centres answered SF-36 and Skindex-29 questionnaires. Control data of validated Dutch population SF-36 scores and literature-acquired scores for other diseases were available. RESULTS: A total of 78 patients were enrolled, of whom 34 (43.6%) were male; the mean age was 39.3 years (SD: 17.1; range: 12-78 years). The Dutch KTS group scored significantly lower than the general Dutch population on all SF-36 scales except Mental Health and Role Emotional. Furthermore, they scored significantly lower than other medical conditions on the Physical Functioning and Bodily Pain scales. According to the Skindex-29 results, KTS patients fall in the categories - symptoms: severe to very severe; emotions: diminutive to mild and functions: mild. The total score is lower than 40, indicating a negligible negative impact on QoL; however, new cut-off values are being calculated. CONCLUSIONS: Classification according to severity is important to educate patients accordingly, predict prognosis and set treatments. Especially in cases of severe KTS, physicians should not only be attentive to the physical aspects but also to the psychological and social aspects of KTS. PMID- 19289310 TI - Performing two DIEP flaps in a working day: an achievable and reproducible practice. AB - BACKGROUND: While the deep inferior epigastric artery perforator (DIEP) flap is a reliable technique for autologous breast reconstruction, the meticulous dissection of perforators may require lengthy operative times. In our unit, we have performed 600 free flaps for breast reconstruction over 8 years and have reduced operative times with a combination of preoperative computed tomographic angiography (CTA), various anastomotic techniques and the Cook-Swartz implantable Doppler probe for perfusion monitoring. We sought to assess the feasibility of performing two DIEP flaps within the working hours of a single day. METHODS: A review of 101 consecutive patients undergoing DIEP flap breast reconstruction in a 12-month period was performed, comparing one DIEP flap per day (n=43) to two DIEP flaps per day (n=58). Complications, outcomes and techniques used were critically analysed. For cases of two DIEP flaps per day, a comparison was made between the use of two separate operating theatres (n=44) and a single consecutive theatre (n=14). RESULTS: Complications did not increase when two DIEP flaps were performed in a single working day. The use of vascular closure staple (VCS) sutures and ring couplers resulted in statistically significant reductions in anastomotic times. The use of two separate theatres for performing two DIEP flaps resulted in a reduction of 59min in operative time per case (p=0.004). CONCLUSION: Two DIEP flaps can be safely and routinely performed within the hours of a single working day. By minimising operative times, these techniques can improve productivity and substantially decrease surgeon fatigue. PMID- 19289311 TI - Children presenting at the emergency department with right lower quadrant pain. AB - Right lower quadrant (RLQ) pain is a common complaint in children presenting at emergency departments. This study analyzed the etiologies of RLQ pain, and compared the clinical presentations, laboratory test results and imaging findings in patients with appendicitis with those in other groups of patients. We also investigated if active observation resulted in delayed diagnosis, to the detriment of patients. Medical records for the period January 2006 to July 2006 were reviewed for children (age < 18 years) who presented to the emergency department of one medical center, complaining of RLQ pain. Out of a total of 100 patients (age range: 2-17 years; mean: 11 years), 46 patients presented with only one symptom of RLQ pain, while 32 patients had >/= 2 associated signs or symptoms, including fever, nausea/vomiting, diarrhea, or rebound tenderness. Imaging studies, including abdominal sonography and/or computed tomography, were performed in 73 patients; 44 underwent surgery for presumed appendicitis and one received surgery for a right paraduodenal hernia. Eleven patients underwent surgery because of peritoneal signs, and eight because of persistent or aggravated RLQ pain. Postoperative pathologic examinations revealed 53 cases of appendicitis, six normal appendices, and other morbidities (1 perforated peptic ulcer, 1 pelvic inflammatory disease, 1 ovarian cyst, 1 diverticulitis, and 1 right paraduodenal hernia). Thirty-three patients were discharged after several hours of observation (range: 0.5-18 hours; mean: 4 hours), and three patients were admitted for further observations. All were discharged without operation. There were significant differences in the incidences of fever (p = 0.004) and rebound tenderness (p = 0.019), and in white cell counts (p < 0.001), neutrophil percentages (p < 0.001), and C-reactive protein levels (p < 0.001) between patients with appendicitis and patients with other causes of RLQ pain. Clinical signs and symptoms, laboratory tests, and imaging studies can be used to differentiate between the causes of RLQ pain. Patients without the classical features of appendicitis or peritonitis can be safely managed by active observation and repeated physical examinations. PMID- 19289312 TI - Effects of the Emergency Trauma Training Course on the confidence of final-year medical students dealing with trauma patients. AB - Trauma is an important issue that has been neglected in the training of medical students. This study evaluated the effects of the Emergency Trauma Training Course (ETTC), after completion of standard medical training, on seventh-year medical students. The ETTC was designed in Taiwan by the Taiwan Society of Emergency Medicine to train physicians and registered nurses who care for trauma patients in the emergency department (ED). We implemented the course for our medical students' internship. One hundred and fifty-one participants were divided into three groups: Group A included 36 medical students before they entered their internship in hospital; Group B included 41 medical students who had received 6 months of internship training in hospital; and Group C included 74 ED nurses. Group C was used to test Cronbach's alpha coefficient of the questionnaire. After the training course, the participants had a final examination and filled out a questionnaire about the training course and their levels of self-confidence. There were no differences in scores between the medical students in Groups A and B (p = 0.064). Using repeated measures analysis of variance, we found that confidence before training was low, with no difference between Groups A and B. Confidence improved after training, but there was still no significant difference between the groups (p = 0.875). However, there were significant differences between confidence levels before and after the training course (p < 0.001). Therefore, although inhospital training for 6 months failed to increase confidence, the confidence of final year medical students after completion of their training was improved by the ETTC. This indicates that the ETTC could increase the confidence of participants. This is the first evaluation of the implementation of the ETTC for final-year medical students in Taiwan. Based on our results, we highly recommend that this training course be taught to final year medical students before they practice in hospital. PMID- 19289313 TI - Reappraisal of the management and outcome of emphysematous pyelonephritis. AB - This study compared the management, prognostic factors and outcomes of patients with emphysematous pyelonephritis (EPN). Twenty-one patients with EPN were studied between September 1996 and August 2005, and were assigned to two groups. Patients in Group 1 received conservative treatment with/without percutaneous catheter drainage (PCD) while patients in Group 2 underwent nephrectomy following medical treatment and PCD. A post hoc analysis of the prognostic factors was performed between survivors and nonsurvivors, and between the survivors in Group 1 and Group 2. There were 14 patients in Group 1, and seven in Group 2. The mortality in Group 1 was 35.7% (5/14) and in Group 2 was 0% (p = 0.12). There were no statistically significant differences in prognostic factors between the two groups, though patients in Group 1 had relatively lower platelet counts (p = 0.07) and Group 2 patients had a higher incidence of dialysis after nephrectomy (p = 0.03). Comparing the survivors and nonsurvivors, patients with comorbid congestive heart failure and patients initially presenting with consciousness disturbances had higher mortalities (p = 0.02 and p < 0.01, respectively). Nonsurvivors also had lower platelet counts (p = 0.06). In conclusion, medical treatment with/without PCD can be used to manage patients with EPN. More agressive drainage is needed in patients with congestive heart failure who initially present with consciousness disturbances or thrombocytopenia. PMID- 19289314 TI - Diabetic muscle infarction in association with terlipressin therapy: a case report. AB - Diabetic muscle infarction (DMI) is a rare complication that often exists in long standing diabetic patients. Clinically, it presents with pain, swelling and sometimes a palpable mass, and is often misdiagnosed as soft tissue infection. The athogenesis of DMI is uncertain. We present the case of a type 2 diabetic patient with liver cirrhosis who was diagnosed with muscle infarction after being treated with terlipressin for gastrointestinal bleeding. This 45-year-old male complained of increasing pain in his right posterior thigh after treatment with terlipressin for 2 days. He was initially diagnosed with soft tissue infection, but he responded poorly to antibiotic treatment. Magnetic resonance imaging suggested acute muscle infarction. We performed a muscle biopsy and the pathologist reported that the muscle was necrotic. After 5 days of bed rest, the patient was able to walk and was discharged uneventfully. PMID- 19289315 TI - Cerebellar hemorrhage after multiple manual pumping tests of a ventriculoperitoneal shunt: a case report. AB - A manual pumping test is commonly used to verify the patency of a ventriculoperitoneal (V-P) shunt. Complications occurring after manual pumping of the reservoir are rare. Here, we report a 73-year-old male hydrocephalic patient who had undergone uneventful V-P shunt surgery 9 years ago. He developed a cerebellar hemorrhage after repetitive pressing of the shunt reservoir. The clinical manifestations, characteristic radiologic images, treatment outcome, and possible mechanisms of this late complication are presented. The risk of massive cerebrospinal fluid overdrainage after repetitive manual pumping of the V-P shunt reservoir, which can contribute to the formation of a cerebellar hemorrhage, warrants special attention and hydrocephalic patients and their caregivers should be informed and educated about this potential complication. PMID- 19289316 TI - Carpal tunnel syndrome and finger movement dysfunction caused by tophaceous gout: a case report. AB - We describe a case of tophaceous gout with a combination of carpal tunnel syndrome and finger movement dysfunction. Carpal tunnel syndrome secondary to gout is uncommon. The concomitant presence of finger movement dysfunction is rare and suggests the involvement of the flexor tendons inside the carpal tunnel. Surgery is recommended to decompress the median nerve, to confirm the diagnosis, and for immediate improvement of flexor tendon excursion. Our patient's finger movement improved dramatically soon after surgery. PMID- 19289317 TI - Coexistence of optic nerve head drusen and combined hamartoma of the retina and retinal pigment epithelium in a Taiwanese male. AB - We report the case of a 38-year-old male patient with a 1-year history of progressively blurred vision in his left eye. He visited our clinic and underwent a series of ophthalmologic examinations including computed tomography, fluorescein angiography and optical coherence tomography. The final diagnosis was optic nerve head drusen and combined hamartoma of the retina and retinal pigment epithelium in his left eye. PMID- 19289318 TI - Gastric subserosal hematoma developing from focal pancreatitis: a case report. AB - Pancreatitis complicated by gastric wall hemorrhagic is rarely reported. The imaging findings of focal pancreatitis with gastric wall hemorrhage can mimic a disease entity of gastric origin. We report a case of gastric subserosal hematoma developing in a patient with focal pancreatitis. The patient was initially scheduled to receive subtotal gastrectomy due to a presumed diagnosis of a disease entity of gastric origin. Computed tomography of the abdomen revealed focal pancreatitis that had eroded the gastric wall resulting in subserosal hemorrhage. The radiologist informed the surgeon of the possibility of focal pancreatitis complicated by gastric hemorrhage. The gastric subserosal hematoma was treated with percutaneous drainage and the patient recovered uneventfully. PMID- 19289319 TI - Impaired orthostatic response in patients with type 2 diabetes mellitus after 48 hours of bed rest. AB - OBJECTIVE: To compare the effect of bed rest on orthostatic responses of patients with type 2 diabetes mellitus and nondiabetic control subjects. METHODS: Six patients with type 2 diabetes and 6 non-diabetic control subjects underwent 48 hours of bed rest and 48 hours of ambulatory activity in randomized order. A 10 minute tilt test was conducted before and after each period of hospitalization, and cardiovascular responses to 80 degrees head-up tilt were analyzed with use of a 2-factorial (study group and bed rest condition) analysis of variance design. We hypothesized that patients with diabetes would experience more severe changes in orthostatic response after bed rest. RESULTS: No significant differences in orthostatic responses were observed before bed rest between control subjects and patients with diabetes. After bed rest, control subjects had a greater (P = .01) increase in heart rate during tilt in comparison with before bed rest (before versus after bed rest, 9 +/- 4 versus 24 +/- 7 beats/min) and maintained their blood pressure during tilt. After bed rest, patients with diabetes did not have a compensatory increase in heart rate and had a greater (P = .02) decline in systolic blood pressure during tilt in comparison with before bed rest (before versus after bed rest, -7 +/- 10 versus -21 +/- 11 mm Hg). Their arm and leg skin vasomotor responses (laser Doppler flowmetry) during tilt were not altered after bed rest and were similar to those in control subjects before and after bed rest. CONCLUSION: Cardiac neuropathy in patients with type 2 diabetes may prevent a compensatory heart rate response after bed rest deconditioning and result in a more severe orthostatic response. A greater decrease in blood pressure with upright tilt is evident after a relatively short period of bed rest. PMID- 19289320 TI - Association between blood glucose levels and development of candidemia in hospitalized patients. AB - OBJECTIVE: To examine the relationship between blood glucose levels in hospitalized patients and the risk of occurrence of candidemia. METHODS: We undertook a retrospective review of medical records and hospital computerized database information to compare blood glucose levels in 48 patients with nosocomial candidemia and 144 contemporaneous matched control subjects without candidemia at a tertiary teaching hospital. RESULTS: The proportions of days (for patients with candidemia versus control subjects without candidemia) with blood glucose levels >or=100 mg/dL (293 of 325 [90%] versus 849 of 1,007 [84%]; P = .009), >or=140 mg/dL (184 of 325 [57%] versus 507 of 1,007 [50%]; P = .049), and >or=200 mg/dL (80 of 325 [25%] versus 163 of 1,007 [16%]; P = .001) were significantly higher during the 7 days preceding the diagnosis of candidemia than during a 7-day period of hospitalization of control subjects. Blood glucose levels exceeding 200 mg/dL for 4 or more days of the week preceding the diagnosis of candidemia were significantly associated with its development (P = .04; odds ratio, 2.44; and 95% confidence interval, 1.01 to 5.94). CONCLUSION: Inpatient hyperglycemia is an important--and potentially modifiable-risk factor for development of nosocomial candidemia. These findings have implications for innovative infection control strategies that focus on glycemic control. PMID- 19289321 TI - Effect of epoprostenol on the thyroid gland: enlargement and secretion of thyroid hormone. AB - OBJECTIVE: To demonstrate the direct stimulation of thyroid tissue in the absence of thyroid-stimulating immunoglobulin after exposure to epoprostenol. METHODS: Seronegative thyrotoxicosis, diffuse goiter, and homogeneous uptake on thyroid scintigraphy were noted in a patient with pulmonary arterial hypertension (PAH) being treated with intravenously administered epoprostenol (prostaglandin I2 or PGI2). More cases with similar characteristics were identified on review of the thyroid function in patients with PAH who were treated with this medication. Fifty-four adult patients with PAH were studied. The study subjects were divided into 2 groups based on whether they were treated with PGI2 or not. Thyroid functions were reviewed, and the prevalence of thyroid disease was assessed. We then compared the prevalence of hyperthyroidism in our study subjects with the prevalence of hyperthyroidism in the general female population using data from published studies. RESULTS: We noted a high prevalence (3 of 45 or 6.7%) of thyroid-stimulating immunoglobulin-negative thyrotoxicosis in adults with preexisting PAH being treated with epoprostenol (PGI2) in the absence of other mechanisms or drugs to explain the hyperthyroidism. The prevalence of hyperthyroidism in our study population was significantly greater (P<.01 by X2 analysis) than that in the general female population in other published reports. CONCLUSION: The data suggest that epoprostenol is a medication associated with stimulation of thyroid tissue, goiter formation, and hyperthyroidism. Patients receiving this drug need to undergo close follow-up for the development of thyrotoxicosis and goiter. PMID- 19289322 TI - Adrenocorticotropic hormone stimulation test during high-dose glucocorticoid therapy. AB - OBJECTIVE: To define the effective time frame of adequate serum cortisol response to the short standard 250-microg adrenocorticotropic hormone stimulation test (ACTH-ST) after initiation of high-dose glucocorticoid therapy in order to assist in the diagnosis of adrenal insufficiency. METHODS: We performed an ACTH-ST at 4 different times in hospitalized patients, who had a documented intact hypothalamic-pituitary-adrenal axis and were receiving high-dose dexamethasone therapy for nonendocrine diseases, to determine the time until the serum cortisol response is compromised. The ACTH-ST was performed at the following time intervals after initiation of dexamethasone therapy - 24, 48, 72, and 96 hours. The outcome measures were cortisol levels measured at 0, 30, and 60 minutes after administration of 250 microg of cosyntropin. RESULTS: Of the overall group of 11 study patients, all those (n = 8) tested within the first 72 hours after initiation of dexamethasone therapy had an adequate response (serum cortisol level at 60 minutes, >or=18 microg/dL). Two of the 3 patients tested at 96 hours after initiation of dexamethasone therapy had a suppressed cortisol response at 60 minutes. At 30 minutes, all cortisol levels except 1 were inadequate (<18 micro/dL). CONCLUSION: This study suggests indirectly that the ACTH-ST may be used for the diagnosis of suspected adrenal insufficiency within the first 3 days after initiation of empiric glucocorticoid therapy. An inadequate response of serum cortisol at 60 minutes to the ACTH-ST may be indicative of adrenal insufficiency in this setting. Additional studies with more patients in each therapy time point are needed to confirm these initial results. PMID- 19289323 TI - Adrenocortical neoplasms with myelolipomatous and lipomatous metaplasia: report of 3 cases. AB - OBJECTIVE: To present pathologic and radiographic features of 3 patients with adrenocortical neoplasms-2 with uncertain malignant potential and 1 adenoma with areas of myelolipomatous and lipomatous metaplasia. METHODS: We describe 3 patients who had adrenocortical neoplasms with foci of myelolipomatous and lipomatous metaplasia. For each patient, the clinical and pathologic data are reviewed. RESULTS: All 3 patients were women, with a mean age at presentation of 58 years. The lesion size averaged 6.8 cm (range, 3.9 to 11.0), and the mean gland weight was 128.8 g (range, 32.5 to 249). Two patients showed imaging findings compatible with adrenal myelolipoma. Pathologically, 2 of the lesions were classified as adrenocortical neoplasms of uncertain malignant potential, and 1 lesion was classified as an adrenocortical adenoma. All 3 lesions contained myelolipomatous foci throughout the neoplasm, and 2 of the tumors contained several pure lipomatous foci. CONCLUSION: Adrenocortical neoplasms, including those associated with an uncertain malignant potential, may be associated with areas of myelolipomatous and lipomatous metaplasia. Imaging studies may result in a false diagnosis of a benign adrenal myelolipoma and potential undertreatment in such patients. PMID- 19289324 TI - Precocious puberty due to rathke cleft cyst in a child. AB - OBJECTIVE: To report a case of a child with precocious puberty attributable to Rathke cleft cyst (RCC). METHODS: The clinical features, laboratory results, and findings on ultrasonography of the pelvis and magnetic resonance imaging of the pituitary gland are presented. RESULTS: A 16-month-old child had breast enlargement, height increase, and an increase in growth velocity. On examination, she was found to have Tanner stage 3 breast development, and her vaginal mucosa was estrogenized. Her height was above the 97th percentile. Biochemically, she was diagnosed as having central precocious puberty, and magnetic resonance imaging of her pituitary gland disclosed RCC. Treatment with leuprolide resulted in normalization of her growth rate and regression of the breast development; the vaginal mucosa also became unestrogenized. CONCLUSION: Although RCC is a relatively common finding, it is a rare cause of precocious puberty. Magnetic resonance imaging of the pituitary gland should be performed in all children younger than 6 years of age who have precocious puberty, in an effort to detect any organic lesions. PMID- 19289325 TI - Silent thyroiditis, isolated corticotropin deficiency, and alopecia universalis in a patient with ulcerative colitis and elevated levels of plasma factor VIII: an unusual case of autoimmune polyglandular syndrome type 3. AB - OBJECTIVE: To describe an unusual case of autoimmune polyglandular syndrome (APS) type 3 and provide a brief review of the literature. METHODS: We present the clinical course and laboratory data of a patient with silent thyroiditis, isolated corticotropin (adrenocorticotropic hormone or ACTH) deficiency, alopecia universalis, and ulcerative colitis with an associated hypercoagulable state. The related literature is also reviewed briefly. RESULTS: A 43-year-old man who had a history of ulcerative colitis with an associated hypercoagulable state and alopecia universalis was referred to the endocrinology department for evaluation of fatigue and a mildly elevated level of thyrotropin (thyroid-stimulating hormone or TSH). He previously had mildly increased TSH levels, for which low dose levothyroxine therapy had been prescribed. During use of this therapy, a suppressed TSH level developed, necessitating discontinuation of thyroid hormone therapy; a subsequent increase in TSH value was followed by a spontaneous return to euthyroidism. An ACTH stimulation test revealed adrenal insufficiency. His ACTH level was low, 21-hydroxylase antibodies were not present, and further testing demonstrated otherwise intact pituitary function. Magnetic resonance imaging of his pituitary gland showed normal findings. Treatment with hydrocortisone promptly decreased his fatigue. He was found to have an elevated factor VIII level as the cause of his hypercoagulable state. The patient continues to feel well with use of hydrocortisone therapy and has normal thyroid function. CONCLUSION: This patient's components of APS type 3 have not been previously reported; thus, the complex nature of the APS variants is supported. PMID- 19289326 TI - Symptomatic hyperprolactinemia from an ectopic pituitary adenoma located in the clivus. AB - OBJECTIVE: To report a case of an ectopic pituitary adenoma in the clivus. METHODS: The clinical, laboratory, and imaging findings of the case are reviewed, and the embryogenesis of the pituitary gland, the pathophysiologic features of this condition, the diagnosis, and the treatment options are discussed. RESULTS: A 20-year-old man presented to a local physician because of a milky nipple discharge of 2 months' duration. He was otherwise healthy. Findings on physical examination were unremarkable except for the milky discharge from both nipples on expression. Serum prolactin, insulinlike growth factor-I, and alpha-fetoprotein were measured. Magnetic resonance imaging of the brain revealed a 13-mm erosive mass in the clivus. Surgical excision of the lesion was undertaken because of the propensity for aggressive lesions in this anatomic location and the high likelihood of complete resection. After the resected tissue was examined, the patient was diagnosed as having an ectopic prolactin-producing pituitary adenoma. His endocrinologic function normalized after resection, and no further therapy was needed. CONCLUSION: Ectopic prolactinoma in the clivus is an uncommon lesion. Surgical resection was undertaken in our patient because of the uncertainty of the diagnosis and the aggressive natural history of more common tumors of the clivus, such as chordomas. Resection provided a cure in this patient. Although it is possible that a successful trial of dopaminergic therapy would have obviated surgical intervention, this approach would be associated with additional risks if the diagnosis were incorrect. PMID- 19289327 TI - Hyperandrogenism due to a testosterone-secreting Sertoli-Leydig cell tumor associated with a dehydroepiandrosterone sulfate-secreting adrenal adenoma in a postmenopausal woman: case presentation and review of literature. AB - OBJECTIVE: To report a case of hyperandrogenism attributable to the presence of an adrenal adenoma secreting dehydroepiandrosterone sulfate (DHEA-S) and an ovarian Sertoli-Leydig cell tumor secreting testosterone in a postmenopausal woman. METHODS: The laboratory, radiologic, and pathologic findings in our case are described. In addition, the pertinent literature is reviewed. RESULTS: A 56 year-old woman presented with a history of gradual increase in facial and body hair, scalp hair loss, male pattern baldness, and deepening of her voice, beginning a few years after spontaneous menopause at age 49 years. She had hypertension, obesity, and type 2 diabetes mellitus. Laboratory tests showed elevated levels of total testosterone (348 ng/dL) and DHEA-S (2,058 microg/dL), and a left adrenal tumor (3 by 4 cm) was detected on abdominal computed tomographic scan. Laparoscopic left adrenalectomy was performed, and the pathologic diagnosis was adrenal adenoma. The DHEA-S returned to normal levels, but the serum testosterone concentration remained elevated. Transvaginal ultrasonography disclosed an ovarian tumor. Bilateral oophorectomy was performed, and an ovarian Sertoli-Leydig cell tumor was diagnosed. The hormonal and clinical picture normalized after this surgical intervention. CONCLUSION: After extensive review of the literature, we believe that this is the first reported case of a coincidental DHEA-S-secreting adrenal adenoma and a testosterone- secreting ovarian Leydig cell tumor causing signs of virilization. PMID- 19289328 TI - Type B insulin resistance developing during interferon-alpha therapy. AB - OBJECTIVE: To report a rare case of diabetes caused by type B insulin resistance due to development of insulin receptor autoantibodies during treatment of hepatitis C with interferon-alpha and ribavirin. METHODS: Clinical and laboratory findings in the case are presented. The literature on type B insulin resistance and interferon-induced autoimmunity is reviewed. RESULTS: A 55-year-old African American man with hepatitis C was treated with interferon and ribavirin. Eight months later, he presented with rapid onset of hyperglycemia, profound weakness, and weight loss. Severe hyperglycemia persisted despite insulin infusion rates as high as 125 U/h. The presence of insulin receptor autoantibodies was confirmed by immunoprecipitation of recombinant human insulin receptor with patient serum. Assays for autoantibodies to islet cell antigens and glutamic acid decarboxylase were negative. The interferon and ribavirin were discontinued. His insulin requirement spontaneously declined to low levels over a 6-month period. Two years after discharge of the patient, insulin receptor autoantibodies could no longer be demonstrated in his serum. He remains euglycemic and is no longer taking insulin. CONCLUSION: This case demonstrates that type B insulin resistance can occur as a complication of interferon-alpha therapy. To our knowledge, this is the first reported case in the United States of type B insulin resistance with development of insulin receptor autoantibodies during treatment with interferon alpha. PMID- 19289329 TI - Neurologic complications of Paget disease of bone. AB - OBJECTIVE: To review the epidemiology, evaluation, and management of the neurologic complications associated with Paget disease of bone (PDB). METHODS: We reviewed the English-language medical literature using MEDLINE data sources from 1950 to August 2008 and manually searched cross-references from original articles and reviews. Search terms included "Paget* disease of bone" and "neurologic* complications," "cranial nerve," "spinal cord," or "peripheral nerve." RESULTS: Several neurologic problems in the central and peripheral nervous systems may complicate PDB. Up to 76% of patients may have some form of neurologic involvement. Neurologic complications can occur in patients with a long history of PDB as well as in patients with previously unrecognized disease. The primary mechanisms of nerve damage in PDB involving the spine are ischemic myelitis and compression due to bone hypertrophy. Evaluation includes determining the serum alkaline phosphatase level and imaging by radiography, bone scintigraphy, computed tomographic scanning, and, for lesions of the central nervous system, magnetic resonance imaging. If a soft-tissue mass is found, biopsy should be considered to exclude the presence of sarcoma. Treatment strategies include calcium, vitamin D, bisphosphonates, and possibly surgical intervention for refractory cases. CONCLUSION: Neurologic sequelae of PDB may be underappreciated. Despite the paucity of data guiding treatment, zoledronic acid is a reasonable first-line therapy. Lack of response to treatment or relapse should prompt diagnostic reevaluation with a heightened suspicion for tumor. PMID- 19289330 TI - Metastatic struma ovarii treated with total thyroidectomy and radioiodine ablation. AB - OBJECTIVE: To present a rare case of metastatic struma ovarii, review the related literature, and discuss the management. METHODS: A case report of a patient with metastatic struma ovarii is presented. The treatment plan, postoperative care, and follow-up are discussed. We conducted a MEDLINE search of the English language literature seeking additional cases of metastatic struma ovarii. In addition to the current case, the previous 40 cases of metastatic struma ovarii were reviewed and analyzed. RESULTS: The mean age of the patients at presentation was 43 years. All patients underwent resection of the primary tumor. The most common sites of involvement for struma ovarii metastatic disease have been the peritoneum, mesentery, and omentum. After primary tumor resection, a wide range of additional treatments have been used, including chemotherapy, resection of metastatic disease, external beam radiation therapy, and radioiodine ablation. CONCLUSION: In cases of metastatic struma ovarii, we recommend total thyroidectomy in conjunction with radioiodine scanning and radioiodine ablation. Thyroglobulin levels should be followed as a tumor marker, and diagnostic radioiodine scans should be performed to screen for residual or recurrent disease. Although this treatment strategy is well established for thyroid cancer, long-term outcomes of this treatment for struma ovarii are still unknown. PMID- 19289331 TI - Giant prolactinoma. PMID- 19289332 TI - Visual vignette. PMID- 19289333 TI - Dextromethorphan abuse: clinical effects and management. AB - OBJECTIVE: To describe the epidemiology, patient presentation, and clinical management associated with dextromethorphan (DM) abuse. DATA SOURCES: PubMed/Medline search using terms dextromethorphan and abuse through July 2008, bibliographies of selected publications, epidemiology tracking databases. STUDY SELECTION: By the authors. DATA EXTRACTION: English language-published review articles, clinical trials, and case reports that described the epidemiologic and toxicologic profile of DM were included. DATA SYNTHESIS: DM is a relatively inexpensive and easily accessible over-the-counter (OTC) medication intended for use as an antitussive. Increasingly, illicit use of the drug has been reported. At clinical doses, the drug produces few adverse effects. However, when abused in large quantities (>2 mg/kg), the drug has been associated with a dissociative effect similar to those described by ketamine and phencyclidine abusers. Massive ingestions of the drug may be associated with untoward effects, including tachycardia, hypertension, and respiratory depression. Overdose symptoms may also be associated with coformulated products such as antihistamines and sympathomimetic amines. Management is primarily supportive. Naloxone has been used to manage DM toxicity but with conflicting reports of effectiveness. CONCLUSION: Recent reports indicate that DM is often abused by individuals seeking its dissociative effects. Clinicians should be aware of the abuse potential of DM. Pharmacists might be particularly cognizant of the risks involved with DM abuse as they control OTC access to the drug. PMID- 19289334 TI - Selective serotonin reuptake inhibitor-induced akathisia. AB - OBJECTIVE: To review available information in the literature about akathisia (inner restlessness) caused by the selective serotonin reuptake inhibitors (SSRIs). DATA SOURCES: Databases searched included Medline, PsychInfo, the International Pharmaceutical Abstracts, and Google Scholar. Search terms included drug-induced akathisia, psychomotor agitation, drug-induced side effect, movement disorders, and extrapyramidal symptoms. These search terms were cross-referenced with selective serotonin reuptake inhibitors and each of the currently marketed SSRIs: fluoxetine, fluvoxamine, sertraline, paroxetine, citalopram, and escitalopram. STUDY SELECTION: Relevant articles were chosen if they specifically mentioned the word akathisia. Case reports were chosen based on a clear view that an SSRI was a contributing or causative agent of akathisia. DATA SYNTHESIS: Recognizing akathisia is important because it can be very bothersome and may cause suicidal ideations. Akathisia can be recognized by examining symptoms, looking at predisposing factors, and using the Barnes Akathisia Rating Scale (BARS). Predisposing factors include use of multiple akathisia-inducing drugs, recent increases in SSRI dose, previous development of akathisia, baseline psychiatric disorders, and brain trauma. Treatment options include the addition of a centrally acting beta-blocker, a benzodiazepine, or an anticholinergic agent. CONCLUSION: Pharmacists can play an active role in recognizing akathisia by being aware of its characteristics, conducting a thorough medication history to identify causative agents, and using BARS to evaluate patients. These efforts may preclude unnecessary discomfort for the patient and reduce the potential for nonadherence induced by akathisia. PMID- 19289337 TI - Jousting with windmills? The quest to achieve our quality aspirations in pharmacy practice. PMID- 19289338 TI - Pharmacists emerge as key stakeholders in quality, patient safety efforts. PMID- 19289339 TI - Improving medication delivery and counseling in community pharmacy. PMID- 19289340 TI - Measurement of pharmacy quality metrics at the pharmacy level should be our goal. PMID- 19289341 TI - Measuring pharmacy quality. AB - OBJECTIVE: To describe methods for measuring health care quality and how these methods can be applied to the measurement of pharmacy quality and to describe ways of stimulating the use of quality improvement methods in pharmacy. SUMMARY: The health care system is moving toward value-based purchasing of professional services, which is also known as value-driven health care. Value is often described as the balance between quality and costs, and thus, we can enhance value by improving quality while controlling costs. Although community pharmacies have not experienced the demand for evidence of quality and value, this is likely to change in the near future as the federal government and private purchasers expand their search for quality-related evidence to all sectors of health care. PQA, a pharmacy quality alliance, has been created to coordinate the efforts of numerous pharmacy stakeholders in developing measures of pharmacy quality and in educating pharmacists about quality improvement methods. In addition to educational strategies for stimulating quality improvement, the pharmacy sector is likely to experience regulatory changes that mandate quality improvement, public reports on the quality of individual pharmacies, and pay-for-performance systems that reward pharmacies for achieving higher levels of quality. CONCLUSION: All stakeholders in pharmacy (i.e., pharmacists, owners, managers, technicians, benefits managers) must become more aware of the movement toward value-driven health care and the ramifications for pharmacy practice. Community pharmacists will soon see an increased demand for evidence of quality and value as this sector is integrated within the broader framework for value-driven health care. PMID- 19289342 TI - Evaluation of a hypertension medication therapy management program in patients with diabetes. AB - OBJECTIVE: To evaluate the effect of a 9-month community pharmacy-based medication therapy management (MTM) program on quality of care in patients with diabetes and hypertension. DESIGN: Randomized controlled trial. SETTING: Tulsa, OK, between November 2005 and July 2007. PATIENTS: 52 patients with diabetes and hypertension who were enrolled in a managed care organization. INTERVENTION: During monthly visits, intervention group participants received MTM services for hypertension and diabetes management. MAIN OUTCOME MEASURES: Systolic blood pressure (SBP), percentage at goal blood pressure (<130/80 mm Hg), and antihypertensive medication adherence. RESULTS: The mean intervention group SBP decreased 17.32 mm Hg, whereas the mean control group SBP level increased 2.73 mm Hg (P = 0.003). The percentage of patients at goal blood pressure increased from 16.0% to 48.0% in the intervention group and decreased from 20.0% to 6.67% in the control group. Intervention group participants were 12.92 times more likely than control group participants to achieve goal blood pressure (P = 0.021). Although the mean adherence rate in the intervention group increased 7.0% while remaining fairly constant in the control group (-0.7%), this group difference was not statistically significant. CONCLUSION: A community pharmacy-based hypertension MTM program was effective in improving blood pressure control among managed care enrollees with diabetes and hypertension. Community pharmacists are strategically positioned to provide MTM services and effectively communicate with providers to improve quality of care for patients. PMID- 19289343 TI - Dispensing errors and counseling quality in 100 pharmacies. AB - OBJECTIVE: To evaluate the dispensing accuracy and counseling provided in community chain pharmacies. DESIGN: Cross-sectional study. SETTING: Community chain pharmacies in large metropolitan areas of Florida, Georgia, New Jersey, and New York. PARTICIPANTS: Community chain pharmacies and trained shoppers. INTERVENTIONS: Trained shoppers presented a new prescription order for one of five study drugs to each randomly selected pharmacy, and all encounters with pharmacy staff were recorded on video by ABC News 20/20 staff using hidden cameras. MAIN OUTCOME MEASURES: Dispensing errors on prescriptions for selected medications were the indicator of prescription dispensing accuracy. Frequency of verbal counseling and information categories discussed or included in written information were used to assess the quality of counseling. RESULTS: Of 100 prescriptions dispensed, 22 had one or more deviation from the physician's written order, for a 22% dispensing error rate. Three of the errors were judged to be potentially harmful when dispensed to a typical patient requiring these therapies. A total of 43 shoppers (43%) received verbal counseling, including 16 cases in which the shopper prompted counseling. All shoppers received written information with their prescription, covering an average of 90% of the required topics. Some 68% of the warfarin shoppers purchased aspirin without the pharmacist verbally warning about taking the drugs simultaneously. CONCLUSION: The dispensing error rate of more than one in five prescriptions is similar to the rate found in a similar study conducted 14 years ago, but counseling frequency has decreased significantly during the period. PMID- 19289344 TI - Strategies to optimize medication use in the physician group practice: the role of the clinical pharmacist. AB - OBJECTIVES: To (1) describe the role of clinical pharmacists in providing population-based pharmaceutical care as employees of a physician group practice, (2) describe the strategies used by pharmacists to optimize medication use, (3) quantify improvements in care, and (4) illustrate the calculations used to quantify cost savings. SETTING: Community-based, multispecialty, physician group practice located in the north Puget Sound area between 2003 and 2007. PRACTICE DESCRIPTION: Using four cornerstones (evidence-based medicine, therapeutic interchange, academic detailing, and a local pharmacy and therapeutics committee), the pharmacists provided population-based pharmaceutical care, leading generic switches, target drug programs, and prescription to over-the counter medication switches. They also led disease management programs, managed drug recalls, implemented electronic health records, negotiated budgets with health plans, and led patient assistance programs and prior authorization programs to improve patient satisfaction. PRACTICE INNOVATION: Implementing these strategies from the vantage point of a physician group presents a seldom-realized employment opportunity for pharmacists. MAIN OUTCOME MEASURES: The impact of these strategies is measured by process, use, and clinical outcomes metrics. These, in turn, are linked to incentive payments in the pay-for-performance environment or to a lowered per member, per month cost in the capitated environment. RESULTS: In 2006-2007, 71% of our hypertensive patients received generic agents compared with a network average for receiving generic agents of 43%, while the proportion of patients with controlled blood pressure increased from 45% to 60%. We saved $450,000 in inpatient costs for deep venous thrombosis. CONCLUSION: Clinical pharmacists employed in a physician group practice can optimize medication use, improve care, and reduce costs. PMID- 19289345 TI - A medication therapy management program's impact on low-density lipoprotein cholesterol goal attainment in Medicare Part D patients with diabetes. AB - OBJECTIVES: To determine a medication therapy management (MTM) service's impact on (1) Healthcare Effectiveness Data and Information Set (HEDIS) quality measures and (2) use and cost expenditures. DESIGN: Nonequivalent group, quasiexperimental study. SETTING: Florida, January 1, 2006, through September 30, 2007. PARTICIPANTS: 2,114 Florida Health Care Plans Medicare Part D enrollees with diabetes. INTERVENTION: Intervention group participated in the MTM program during the HEDIS measurement year. MAIN OUTCOME MEASURES: Presence of low-density lipoprotein cholesterol (LDL-C) screening, LDL-C values, and LDL-C control (<100 mg/dL). The use measure was the total number of 30-day medication equivalents. Cost measures were (1) total Medicare Part D drug cost, (2) enrollees' out-of pocket Part D medication costs, and (3) total medication copayments. Statistical analyses included chi-square, independent and paired t tests, and analysis of variance with post hoc comparisons. RESULTS: Of 2, 114 enrollees eligible for comprehensive diabetes care (CDC) according to HEDIS guidelines, 255 participated in the MTM intervention group and 56 patients were MTM eligible but opted out of the program or could not be reached for medication review during 2008 (MTM nonparticipants). A higher proportion of patients in the MTM participant group had LDL-C levels less than 100 mg/dL (69.0%) compared with those in the MTM nonparticipant (50.0%) and CDC only (54.1%) groups (chi2 = 20.9(3), P < 0.001). The two control groups' average LDL-C (90.8 and 93.6 mg/dL) was significantly higher than the intervention group (83.4 mg/dL, P < 0.001). Overall, per member per month use and drug costs differed from 2007 to 2008 and enrollees in the MTM participant group had greater percentage cost reductions. CONCLUSION: Enrollees who were eligible for MTM services but did not receive them had poorer clinical, use, and cost outcomes compared with the MTM intervention group. Pharmacists collaborating with physicians through a MTM program can improve quality of metrics for chronic diseases and reduce medication costs. PMID- 19289346 TI - Clinical and economic impact of a diabetes clinical pharmacy service program in a university and primary care-based collaboration model. AB - OBJECTIVE: To provide program methodology and outcomes data identifying the impact of clinical pharmacy services (CPSs) in patients with type 2 diabetes. DESIGN: Longitudinal pre-post cohort study. SETTING: Regional primary care group in Buffalo, NY, during 2006-2007. PATIENTS: Patients with type 2 diabetes identified by their primary care providers were referred to the MedSense program; a pharmacist-led, patient-centered pharmacotherapy management program developed through university collaboration with a regional primary care physician group. INTERVENTIONS: Education, clinical assessments, provider recommendations, and longitudinal follow-up of treatment goals provided by MedSense pharmacists. MAIN OUTCOME MEASURES: Clinical outcomes were followed for 1 year from the index date for primary diabetes endpoints (glycosylated hemoglobin and fasting plasma glucose) and accompanying metabolic parameters (body mass index, blood pressure, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglycerides). Economic endpoints from the payer perspective were also followed for 1 year from the index date for medical and prescription-related costs. RESULTS: Primary diabetes endpoints were significantly reduced versus baseline at the 6-month (-1.1%; P < 0.0001, -39 mg/dL; P = 0.003) and 12-month (-1.1%; P < 0.0001, -35 mg/dL; P = 0.005) assessments. Improvement rates were observed for all accompanying metabolic parameters at each assessment (range 40-64%). Geometric mean costs tended to decrease versus baseline at 6-month (-$84; P = 0.785) and 12-month (-$216; P = 0.414) assessments, despite nominal increases in diabetes and total medication costs. CONCLUSION: In this CPS model, there were initial and sustained reductions in the primary diabetes endpoints and a high rate of improvement for accompanying metabolic parameters. Concurrent with clinical improvements, total direct medical costs were reduced despite an increase in antidiabetic medication and total medication costs. PMID- 19289347 TI - Use of community pharmacy-provided diabetes services to aid physicians in the National Committee for Quality Assurance recognition program. AB - BACKGROUND: Pay-for-performance (P4P) models are being adopted by many health care payers, including Medicare, for payment of physician services. To receive financial incentives in P4P programs, physicians are encouraged to attain recognition or credentialing from an agency, such as the National Committee for Quality Assurance (NCQA). OBJECTIVE: To explore the potential roles of a community pharmacy-provided diabetes services in collaboration with physicians and to assist them in becoming acknowledged by the NCQA Diabetes Physician Recognition Program. SUMMARY: KDI Health Solutions pharmacists have demonstrated success in the management of diabetes through the Asheville Project and the Diabetes Ten City Challenge. Continued pharmacists' efforts may focus on expanding current disease state management programs, community pharmacy diabetes education programs, and collaborative practices with physicians. Studies are currently being conducted to assess physician willingness to use pharmacists as part of the NCQA recognition process. CONCLUSION: Collaboration of physicians and pharmacists may improve diabetes standards of care and aid physicians in becoming NCQA-recognized providers. NCQA-recognized physicians can align themselves with current and future P4P programs. PMID- 19289348 TI - Development and testing of performance measures for pharmacy services. AB - OBJECTIVE: To report on the status of the pilot work of PQA, a pharmacy quality alliance, to develop and test performance metrics of pharmacy services for use in quality improvement, benchmarking, and pay-for-performance benchmarks. DESIGN: Observational cohort study. SETTING: Three health plans (commercial, Medicare and Medicaid) located in the northeastern United States and one nationwide prescription drug plan. PATIENTS: Pharmacies of health plans with membership ranging from approximately 3,330 to nearly 1.7 million members. INTERVENTION: Pharmaceutical claims data for prescriptions dispensed at community pharmacies were analyzed. MAIN OUTCOME MEASURES: Not applicable. RESULTS: The four plans had pharmacy networks ranging from 653 to 53,153 pharmacies. When using a minimum sample of 30 members per measure, less than 10% of the pharmacies within the plans' networks were evaluable for all measures except the measure of high-risk drugs in the elderly. The measure for high-risk drugs in the elderly had 6,210 evaluable pharmacies in a network of 53,153. The measures for high-risk drugs in the elderly and medication adherence appear to have the greatest potential for use as performance measures in that they show room for improvement and variation among pharmacies. CONCLUSION: The ideal performance measure is relevant, scientifically sound, and feasible. Several of the measures that underwent testing possessed some, if not all, of the properties of an ideal performance measure. Strategies for aggregating data across health and drug plans may be useful for overcoming sample size challenges. PMID- 19289349 TI - Demonstration of community pharmacy and managed care organization collaboration on cardiovascular disease risk factor identification using health risk appraisal. AB - OBJECTIVE: To integrate the resources from a local statewide managed care organization (MCO) and a supermarket pharmacy chain to conduct a comprehensive health risk appraisal (HRA). Collected data were used to assess cardiovascular risk factors and identify disease management opportunities. METHODS: An analysis to determine the prevalence of risk factors was conducted on a cross-sectional HRA survey. The HRA involved point-of-care cholesterol screening (with a follow up risk factor questionnaire) conducted by pharmacist employees of the employer group (a regional supermarket chain). Those eligible for the screening were employees of the supermarket chain and their dependents covered by the participating MCO. RESULTS: A total of 12,915 completed HRA questionnaires were received. The mean age of the employees participating was approximately 44 years. Of note, 14%, 24%, 21%, and 69% of questionnaires had abnormal values for total cholesterol, high-density lipoprotein (HDL) cholesterol, blood pressure, and body mass index (BMI), respectively. Compared with national benchmarks, low HDL cholesterol and BMI more than 30 kg/m2 were more common in this cohort. More than one-fourth of the employees in this analysis were identified as being at high risk for a coronary heart disease event. CONCLUSION: The unique collaboration presented here allowed for an expanded role of pharmacists to implement a quality improvement program. In response, the employer decided to continue the HRA screening and offer a employee contribution reduction-based health incentive to covered members. The employer also is considering offering cardiovascular disease management interventions that will be performed by the supermarket chain's pharmacists and targeted toward the identified risk factor trends. PMID- 19289350 TI - Glucose dysregulation among veterans living with schizophrenia-related disorders after switching second-generation antipsychotics. AB - OBJECTIVES: To compare (1) blood glucose and glycosylated hemoglobin (A1C) laboratory results and (2) longitudinal trends in blood glucose levels among veterans switched from one second-generation antipsychotic (SGA) to another. DESIGN: Retrospective, naturalistic, nonequivalent control group. SETTING: United States between April 1, 2003, and September 30, 2003. PATIENTS: 1,776 U.S. Veterans Health System beneficiaries living with schizophrenia-related disorders switching (1) from olanzapine to another SGA, (2) to olanzapine from another SGA, and (3) among nonolanzapine SGAs. INTERVENTION: Data were retrieved from the laboratory results (LAR) database for a maximum of 180 days before and 365 days after the index date. MAIN OUTCOME MEASURES: Mean blood glucose, A1C, and change in blood glucose. RESULTS: Blood glucose (36.0 mg/dL, paired t test109 = -4.87, P < 0.001) and A1C (1.0%, paired t143 = -4.84, P < 0.001) declined among veterans switched from olanzapine who were taking a blood glucose-lowering agent before the switch but was unchanged for those who were not. Adjusting for age, gender, and race, addition of the switch-type variables improved prediction of blood glucose change (F-ratio = 3.76, P = 0.03). Linear mixed-effects models confirmed that blood glucose levels declined for veterans switched from olanzapine with glucose dysregulation before the switch (Est(beta2 - beta1) = -34.5 mg/dL, t424 = -5.05, P < 0.001). CONCLUSION: Blood glucose and A1C were significantly improved among veterans switched from olanzapine with evidence of glucose dysregulation before the switch. They were stable among those without evidence of preexisting glucose dysregulation. Therapeutic switches from one SGA to another should be monitored as a risk factor for changes in glucose regulation. PMID- 19289351 TI - Development of a medication therapy management superbill for ambulatory care/community pharmacy practice. AB - OBJECTIVES: To explain the purpose of superbills, suggest strategies for incorporating superbills into pharmacy practice, and propose a model superbill for consideration by practitioners. PRACTICE DESCRIPTION: Ambulatory pharmacies in the United States. PRACTICE INNOVATION: Superbills have been used by physicians and other health care providers for many years as a way of efficiently communicating to the office staff, the patient, and even the insurer the types of services that have been provided at the point of care. The profession of pharmacy has not routinely used superbills in the past; however, given the recognition of pharmacists as providers of medication therapy management (MTM) services, immunizations, disease management, and other specialty preventive health services, the time has come for pharmacists to begin using superbills. MAIN OUTCOME MEASURES: Not applicable. RESULTS: A sample superbill, suitable for adaptation by individual providers of medication therapy management and other clinical pharmacy services, is provided in this article. CONCLUSION: Superbills may or may not improve the pharmacist's overall ability to receive insurance remuneration, but the authors believe that greater recognition by patients of the nondispensing activities of pharmacists can be achieved by using a superbill and that this may lead to more opportunities for payment for MTM in the future. Research is needed to assess whether incorporating superbills into a variety of pharmacy practice settings improves patient perceptions of the pharmacist and to discover how superbills effect practice efficiency. PMID- 19289353 TI - John Uri Lloyd 1849-1936: wizard of American plant pharmacy. PMID- 19289354 TI - New drugs: milnacipran hydrochloride, fesoterodine fumarate, and silodosin. PMID- 19289356 TI - Lateralized repetition priming for unfamiliar faces. AB - Repetition priming (RP) is the ability to recognize a stimulus more rapidly as a result of prior exposure to the item. Recent research examining the neuroanatomical basis of this effect has demonstrated RP for familiar faces presented to the right but not to the left cerebral hemisphere. Extending this line of enquiry, the current research considered whether similar effects emerge when unfamiliar faces are the stimuli of interest. Using a divided-visual-field methodology, RP for unfamiliar faces in the left and the right hemispheres was assessed. The results revealed that RP: (i) only emerges in the right hemisphere; (ii) is evident regardless of whether the lateralized presentation of unfamiliar faces occurs at study or at test and (iii) occurs only when hair is cropped from the faces. The theoretical implications of these findings are considered. PMID- 19289357 TI - On the crucial role of mental ingroup representation for ingroup bias and the ingroup prototypicality-ingroup bias link. AB - Previous research suggested that relative ingroup prototypicality is a basis for ingroup bias. To test the boundary conditions of this phenomenon, we hypothesized that people particularly rely on relative ingroup prototypicality as a basis for ingroup bias if the prototypicality information is derived from a homogeneous and simple ingroup representation. We, therefore, predicted increased ingroup bias together with a stronger relation between prototypicality and ingroup bias if the ingroup is formed of consistent group members only. In two experiments, we used different subtyping manipulations and showed that the exclusion of inconsistent parts of the ingroup leads to a strong relation between relative ingroup prototypicality and ingroup bias, whereas this relation was nonsignificant without subtyping. Furthermore, ingroup bias was more pronounced after subtyping. These results confirm that the homogeneity and the simplicity of the ingroup representation is an important moderator for the relation between ingroup projection and intergroup judgments. PMID- 19289358 TI - Automatic selection of irrelevant object features through working memory: evidence for top-down attentional capture. AB - Recent research has shown that the contents of working memory (WM) can guide the early deployment of attention in visual search. Here, we assessed whether this guidance occurred for all attributes of items held in WM, or whether effects are based on just the attributes relevant for the memory task. We asked observers to hold in memory just the shape of a coloured object and to subsequently search for a target line amongst distractor lines, each embedded within a different object. On some trials, one of the objects in the search display could match the shape, the colour or both dimensions of the cue, but this object never contained the relevant target line. Relative to a neutral baseline, where there was no match between the memory and the search displays, search performance was impaired when a distractor object matched both the colour and the shape of the memory cue. The implications for the understanding of the interaction between WM and selection are discussed. PMID- 19289359 TI - Translation priming between the native language and a second language: new evidence from Dutch-French bilinguals. AB - During the last two decades, bilingual research has adopted the masked translation priming paradigm as a tool to investigate the architecture of the bilingual language system. Although there is now a consensus about the existence of forward translation priming (from native language primes (L1) to second language (L2) translation equivalent targets), the backward translation priming effect (from L2 to L1) has only been reported in studies with bilinguals living in an L2 dominant environment. In a lexical decision experiment, we obtained significant translation priming in both directions, with unbalanced Dutch-French bilinguals living in an L1 dominant environment. Also, we demonstrated that these priming effects do not interact with a low-level visual prime feature such as font size. The obtained backward translation priming effect is consistent with the model of bilingual lexicosemantic organization of Duyck and Brysbaert (2004), which assumes strong mappings between L2 word forms and underlying semantic representations. PMID- 19289360 TI - Of colored numbers and numbered colors: interactive processes in grapheme-color synesthesia. AB - Grapheme-color synesthetes experience a specific color when they see a grapheme but they do not report to perceive a grapheme when a color is presented. In this study, we investigate whether color can still evoke number-processes even when a vivid number experience is absent. We used color-number and number-color priming, both revealing faster responses in congruent compared to incongruent conditions. Interestingly, the congruency effect was of similar magnitude for both conditions, and a numerical distance effect was present only in the color-number priming task. In addition, a priming task in which synesthetes had to judge the parity of a colored number revealed faster responses in parity congruent than in parity incongruent trials. These combined results demonstrate that synesthesia is indeed bi-directional and of similar strength in both directions. Furthermore, they illustrate the precise nature of these interactions and show that the direction of these interactions is determined by task demands, not by the more vividly experienced aspect of the stimulus. PMID- 19289361 TI - Strengthening the case for stimulus-specificity in artificial grammar learning: no evidence for abstract representations with extended exposure. AB - Different theories have been proposed regarding the nature of the mental representations formed as a result of implicit learning of sequential regularities. Some theories postulate abstract surface-independent representations, while other theories postulate stimulus-specific representations. This article reports three experiments investigating the development of abstract representations in artificial grammar learning (AGL), using a methodological approach developed by Conway and Christiansen (2006). In all the experiments, the number of blocks during the exposure phase was manipulated (6 blocks vs. 18 blocks of exposure to sequences). Experiments 1 and 2 investigated both visual and auditory learning where sequences were presented element-by-element. Experiment 3 investigated visual learning using a sequence-by sequence presentation technique more commonly used in visual AGL studies. Extending previous research (Conway & Christiansen, 2006) and in support of stimulus-specific accounts, the results of the experiments showed that extended observational learning results in increased stimulus-specific knowledge rather than abstraction towards surface-independent representations. PMID- 19289362 TI - Like me or like us: is ingroup projection just social projection? AB - Research has shown abundant evidence for social projection, that is, the tendency to expect similarity between oneself and others (Krueger, 1998a, 1998b). This effect is stronger when others constitute an ingroup instead of an outgroup (Robbins & Krueger, 2005). A different line of research has shown evidence for ingroup projection, that is, the tendency to use ingroup instead of outgroup features to define an inclusive category (Wenzel, Mummendey, Weber, & Waldzus, 2003). In this research, we examine whether ingroup (i.e., Germans) projection to an inclusive category (i.e., Europeans) is different from the projection of self attributes to the same inclusive category. In two studies, German participants rated how typical a series of attributes was for the ingroup, an inclusive category, the self, and an outgroup (i.e., Italians). The attributes varied in their relevance to the groups under consideration. The results showed that ingroup projection is stronger than social projection, but only when typical ingroup attributes are concerned. Moreover, ingroup projection weakened when the image of the ingroup was manipulated to be less positive, while social projection remained stable. These findings imply that ingroup projection is not simply social projection. PMID- 19289363 TI - Conflict-frequency affects flanker interference: role of stimulus-ensemble specific practiceand flanker-response contingencies. AB - Performance in choice reaction time tasks deteriorates when an irrelevant stimulus feature is associated with an incorrect response (conflict condition). Such interference effects are reduced under conditions of increased conflict frequency. Although models of cognitive control account for this modulation in terms of conflict-related attentional focusing on the target stimulus dimension, it is possible that the effect reflects practice with specific stimulus ensembles or stimulus feature-response contingencies. Using an Eriksen flanker task, we deconfounded the frequency of conflict trials and the frequency of specific stimulus ensembles (i.e., target-flanker conjunctions). In Experiments 1 and 2, flanker interference varied inversely with the frequency of conflict trials, irrespective of practice with specific target-flanker conjunctions, thereby disputing a stimulus ensemble learning account. In Experiment 3, however, flanker interference was reduced for specific flanker stimuli which occurred predominantly in conflict trials. Taken together, the findings are consistent with flanker-specific attentional adjustment or associative flanker-response priming. PMID- 19289365 TI - Circulating MicroRNA Signatures of Tumor-Derived Exosomes for Early Diagnosis of Non-Small-Cell Lung Cancer. PMID- 19289366 TI - Targeted therapy in lung cancer: the good, the bad, and the ugly. PMID- 19289368 TI - Beyond doublet chemotherapy for advanced non-small-cell lung cancer: combination of targeted agents with first-line chemotherapy. AB - The first-line treatment of advanced non-small-cell lung cancer (NSCLC) has evolved significantly over the past 5 years. As recently as 15 years ago, best supportive care (BSC) was considered an acceptable option for most patients with advanced or metastatic NSCLC, based on the concern that toxic effects of systemic chemotherapy overshadowed any potential benefits. The enhanced efficacy of platinum-based doublet chemotherapeutic regimens led to increases in overall patient survival relative to BSC. However, overall survival (OS) appeared to plateau, even with the introduction and refinement of these regimens. The addition of novel targeted agents targeting growth pathways to platinum-based regimens failed to overcome the 7.8- to 10.5-month survival barrier. After many phase III clinical trials, which involved tyrosine kinase inhibitors, matrix metalloproteinase inhibitors, protein kinase C inhibitors, and retinoids, this survival barrier had yet to be surmounted, although in some cases certain subgroups benefited, suggesting specific molecular correlations. Recently, inhibition of components of the angiogenesis pathway with the addition of bevacizumab to a platinum-based doublet led to statistically significant increases in OS, progression-free survival, and response rate relative to chemotherapy alone. This advance pushed the median survival of selected patients with advanced or metastatic NSCLC who met the eligibility criteria of the trial over the 12-month mark, thus offering patients and clinicians hope for more incremental advances in the future. PMID- 19289369 TI - Toxicity of targeted therapy in non-small-cell lung cancer management. AB - Lung cancer is the leading cause of cancer-related death worldwide. Despite several chemotherapeutic agents, a survival plateau has been reached, so new treatment strategies are clearly needed. A strong interest is now focused on the use of targeted therapies for the management of non-small-cell lung cancer. Monoclonal antibodies against the epidermal growth factor receptor (EGFR; cetuximab) or vascular endothelial growth factor receptor (VEGFR; bevacizumab) and EGFR tyrosine kinase inhibitors (gefitinib, erlotinib) are generally well tolerated and do not have the severe systemic side effects usually seen with cytotoxic drugs. A considerable number of treated patients develop dermatologic side effects, such as acneiform eruption, xerosis, and eczema, and unfortunately, this is often one cause of negative impact on a patient's quality of life. No controlled clinical trials have been performed to manage rash, so it is necessary to provide suggestions for managing this frequent side effect. The main problems related to the class of angiogenesis inhibitors affecting VEGFRs are the exclusion of patients with brain metastases and/or squamous histology, and vascular adverse effects, such as hypertension, proteinuria, thrombosis, and hemorrhage. There are other new agents in clinical development, such as sorafenib, sunitinib, vorinostat, vandetanib, everolimus, panobinostat, and ASA404. They are all associated with a spectrum of toxicities, often reversible with interruption of dosing. Further research is required to clarify the role of targeted therapies and toxicities management. PMID- 19289370 TI - Neoadjuvant and intrapleural therapies for malignant pleural mesothelioma. AB - Historically, malignant pleural mesothelioma patients with potentially resectable disease have been treated with surgery and radiation alone. With improvements in systemic and intrapleural treatment options, a movement toward multi-modality therapy has become more common. Systemic treatment options largely consist of neoadjuvant chemotherapy with platinum doublets and most recently novel targeted agents, such as dasatinib. Intrapleural strategies have included injecting chemotherapy, chemotherapy with hyper thermic per fusion, gene therapy, and immunotherapy. The following review discusses the latest results in neoadjuvant and intrapleural therapies in malignant pleural mesothelioma. PMID- 19289371 TI - Exosomal microRNA: a diagnostic marker for lung cancer. AB - PURPOSE: To date, there is no screening test for lung cancer shown to affect overall mortality. MicroRNAs (miRNAs) are a class of small noncoding RNA genes found to be abnormally expressed in several types of cancer, suggesting a role in the pathogenesis of human cancer. PATIENTS AND METHODS: We evaluated the circulating levels of tumor exosomes, exosomal small RNA, and specific exosomal miRNAs in patients with and without lung adenocarcinoma, correlating the levels with the American Joint Committee on Cancer (AJCC) disease stage to validate it as an acceptable marker for diagnosis and prognosis in patients with adenocarcinoma of the lung. RESULTS: To date, 27 patients with lung adenocarcinoma AJCC stages I-IV and 9 controls, all aged 21-80 years, were enrolled in the study. Small RNA was detected in the circulating exosomes. The mean exosome concentration was 2.85 mg/mL (95% CI, 1.94-3.76) for the lung adenocarcinoma group versus 0.77 mg/mL (95% CI, 0.68-0.86) for the control group (P < .001). The mean miRNA concentration was 158.6 ng/mL (95% CI, 145.7-171.5) for the lung adenocarcinoma group versus 68.1 ng/mL (95% CI, 57.2-78.9) for the control group (P < .001). Comparisons between peripheral circulation miRNA derived exosomes and miRNA-derived tumors indicated that the miRNA signatures were not significantly different. CONCLUSION: The significant difference in total exosome and miRNA levels between lung cancer patients and controls, and the similarity between the circulating exosomal miRNA and the tumor-derived miRNA patterns, suggest that circulating exosomal miRNA might be useful as a screening test for lung adenocarcinoma. No correlation between the exosomal miRNA levels and the stage of disease can be made at this point. PMID- 19289372 TI - XPG mRNA expression levels modulate prognosis in resected non-small-cell lung cancer in conjunction with BRCA1 and ERCC1 expression. AB - BACKGROUND: Molecular markers can help identify patients with early-stage non small-cell lung cancer (NSCLC) with a high risk of relapse. Excision repair cross complementing 1 (ERCC1), Xeroderma pigmentosum group G (XPG), and breast cancer 1 (BRCA1) are involved in DNA damage repair, whereas ribonucleotide reductase M1 (RRM1) is implicated in DNA synthesis. Expression levels of these molecules might therefore have a prognostic role in lung cancer. PATIENTS AND METHODS: We examined ERCC1, RRM1, XPG, and BRCA1 mRNA levels by real-time quantitative polymerase chain reaction in 54 patients with stage IB-IIB resected NSCLC. A strong correlation was observed between the 4 genes. RESULTS: For patients with low BRCA1, regardless of XPG mRNA expression levels, disease-free survival (DFS) was not reached. For patients with intermediate/high BRCA1 and high XPG, DFS was 50.7 months. However, for patients with intermediate/high BRCA1 and low/intermediate XPG, DFS decreased to 16.3 months (P = .002). Similar differences were observed in overall survival, with median survival not reached for patients with low BRCA1, regardless of XPG levels, or for patients with intermediate/high BRCA1 and high XPG. Conversely, for patients with intermediate/high BRCA1 levels and low/intermediate XPG levels, median survival dropped to 25.5 months (P = .007). CONCLUSION: BRCA1 and XPG were identified as independent prognostic factors for both median survival and DFS. High BRCA1 mRNA expression confers poor prognosis in early NSCLC, and the combination of high BRCA1 and low XPG expression still further increases the risk of shorter survival. These findings can help optimize the customization of adjuvant chemotherapy. PMID- 19289373 TI - Cisplatin plus gemcitabine as adjuvant chemotherapy for radically resected non small-cell lung cancer: a pilot study. AB - BACKGROUND: The combination cisplatin/gemcitabine is one of the most active and well-tolerated regimens in advanced non-small-cell lung cancer (NSCLC). We undertook this pilot study to evaluate postoperative drug delivery and toxicity of cisplatin plus gemcitabine in patients with radically resected stage IB-III NSCLC. PATIENTS AND METHODS: Twenty-two consecutive patients were treated with cisplatin 80 mg/m2 on day 1 and gemcitabine 1200 mg/m2 on days 1 and 8 every 3 weeks for 4 planned courses. Most patients (50%) had pathologic stage IIIA disease; all had an Eastern Cooperative Oncology Group performance status score of 0 or 1. The median age was 63 years (range, 56-73 years). Six out of 22 patients (27.3%) had undergone pneumonectomy. RESULTS: A total of 85 courses of chemotherapy were administered, and the median number of courses was 4 (range, 1 4 courses); twenty-one out of 22 patients received the planned 4 courses of chemotherapy. Chemotherapy caused grade 3 anemia in 1 patient, grade 3 thrombocytopenia in another patient, and grade 3/4 neutropenia in 4 patients; grade 3 vomiting was observed in 3 patients, and grade 3 anorexia and grade 3 asthenia were both observed in 1 patient. No treatment-related deaths occurred. The median delivered dose intensities of cisplatin and gemcitabine were 24.3 mg/m2/week (97.2%) and 700.9 mg/m2 weekly (87.5%), respectively. CONCLUSION: The combination of cisplatin/gemcitabine is a feasible and well-tolerated regimen in the adjuvant setting. Future trials should better define the best strategy in terms of efficacy, toxicity, and quality of life between this combination and the standard regimen, cisplatin plus vinorelbine. PMID- 19289374 TI - Eukaryotic initiation factor-4E and cyclin D1 expression associated with patient survival in lung cancer. AB - Eukaryotic initiation factor 4E (eIF4E) and cyclin D1, two important factors in cell-cycle progression, play key roles in the carcinogenesis of varied human cancers. However, eIF4E expression in non-small-cell lung cancer (NSCLC) and its association with cyclin D1 has received little investigation. One hundred forty seven subjects with primary NSCLC, with long-term follow-up and essential clinicopathologic parameters (including age, sex, tumor grade, tumor stage, smoking history, performance status, weight loss, histology grade, and survival data) were evaluated based on expression of eIF4E and cyclin D1. Immunohistochemical analysis was performed using monoclonal antibodies against eIF4E and cyclin D1. While 134 of 147 cases (91%) were positive for eIF4E, 82 of 136 cases (63%) were positive for cyclin D1. Western blot results were consistent with those illustrated by immunohistochemistry. While eIF4E(+) correlated with significantly shorter patient survival (P = .03), cyclin D1(+) correlated with longer patient survival (P = .01). Assessment of coexpression of cyclin D1 and eIF4E shows greater value in determining the prognosis of NSCLC: patients with eIF4E(+)/cyclin D1(-) have poorer outcome, those with eIF4E(-)/cyclin D1(+) have a more favorable outcome, and those with eIF4E(+)/cyclin D1(+) have an intermediate outcome (P = .02). The negative effect on survival in patients with eIF4E(+) suggests its potential prognostic role in NSCLC. These results warrant further investigation to explore the value of eIF4E in identifying patients with aggressive disease for adjuvant treatments. PMID- 19289376 TI - Complementary therapy use and health self-management among rural older adults. AB - OBJECTIVES: This article describes dimensions of complementary therapy use among rural older adults, employs these dimensions to delineate sets of complementary therapy use, and describes the personal characteristics related to each set of complementary therapy use. METHODS: Data are from in-depth interviews conducted with 62 African American and White rural older adults. RESULTS: Three dimensions of complementary therapy use are delineated: types of therapies used, mindfulness in therapy use, and sharing information with conventional health care providers. The intersection of these dimensions indicates 5 patterned sets of complementary therapy use among rural older adults: (a) mindful use of only home remedies; (b) mindful use of home remedies and contemporary supplements; (c) mindful use of home remedies, contemporary supplements, and complementary practices; (d) nonmindful use of home remedies and contemporary supplements; and (e) use of conventional care only. Involvement in the 5 sets of therapy use is related to sex, ethnicity, educational attainment, and migration. DISCUSSION: Understanding how older adults include sets of complementary therapies in their health self management is important for improving their health care resources, expectations, awareness, and priorities. PMID- 19289377 TI - Impaired vascular function in small resistance arteries of LOX-1 overexpressing mice on high-fat diet. AB - AIMS: LOX-1 is a major vascular receptor for oxidized low-density lipoprotein (oxLDL). In this study, we analysed the impact of LOX-1 overexpression and high dietary fat intake on vascular function in small resistance arteries. METHODS AND RESULTS: Relaxation of mesenteric arteries was measured using a wire myograph. Compared with the control group, mice overexpressing LOX-1 on a high-fat diet (FD) had preserved vascular smooth muscle relaxation, but impaired endothelium dependent relaxation via NO. Vascular NO availability was decreased by exaggerated formation of reactive oxygen species and decreased endothelial NO synthase expression. Endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation via cytochrome P450 metabolites was increased in LOX-1 + FD animals, but did not completely compensate for the loss of NO. Currents of calcium activated potassium channels with large conductance (BKCa channels) were measured by the voltage-clamp method. The BKCa current amplitudes were not altered in endothelial cells, but highly increased in vascular smooth muscle cells from resistance arteries of LOX-1-overexpressing mice on FD. BK(Ca) currents were activated by low-dose H2O2 and cytochrome P450 metabolites 11,12-EET and 14,15 EET as EDHF in control mice. CONCLUSION: LOX-1 overexpression and FD caused functional changes in endothelial and vascular smooth muscle cells of small resistance arteries. PMID- 19289381 TI - The effect of obesity on intrahepatic cytokine and chemokine expression in chronic hepatitis C infection. AB - BACKGROUND: Obese subjects with chronic hepatitis C virus (HCV) infection have more rapidly progressive liver disease. Objective In this study, we aimed to compare the hepatic cytokine and chemokine profiles in obese and lean subjects with chronic HCV infection using qRT-PCR and immunohistochemistry. METHODS: Liver biopsies from 55 subjects were studied, including 20 with chronic hepatitis C, 25 with non-alcoholic fatty liver disease and 10 subjects with non-diseased liver. RESULTS: Compared to the control groups, the liver injury in chronic hepatitis C was characterised by increased expression of several T-helper-1 cytokines including interferon-gamma and tumour necrosis factor-alpha, and chemokines such as RANTES, IP-10 and MCP-1. In particular, in comparison with lean (BMI or=30 kg/m(2)) HCV infected subjects had increased hepatic expression of interferon-gamma (p=0.004) and tumour necrosis factor-alpha (p<0.001), as well as increased expression of IP-10 (p=0.009) and MCP-1 (p<0.001). Localisation of these inflammatory chemokines revealed that in comparison to lean-HCV subjects, HCV infected liver from obese subjects exhibited significantly increased expression of IP-10 (p<0.001) and MCP 1 (p=0.02) in the inflammatory infiltrate of the portal tracts. In parallel, there was increased CD3 infiltration in the liver of obese-HCV subjects. CONCLUSIONS: The data provide important mechanistic information on the cause of hepatic injury in obese-HCV subjects including: (1) enhanced T helper-1 cytokine response patterns-to promote hepatocellular injury; (2) increased expression of the chemokines IP-10 and MCP-1 at both the mRNA and protein levels-to enhance inflammatory cell recruitment; (3) differing localisation of these chemokines within the liver of obese-HCV versus lean-HCV subjects-implying different inducing stimuli and; (4) increased CD3 expression in the liver of obese-HCV subjects-concordant with the increased expression of T cell chemoattractants. PMID- 19289382 TI - Does anti-mutated citrullinated vimentin have additional value as a serological marker in the diagnostic and prognostic investigation of patients with rheumatoid arthritis? A systematic review. AB - OBJECTIVE: To review the diagnostic and prognostic value of anti-mutated citrullinated vimentin (MCV) in rheumatoid arthritis, taking into account the already available serology. METHODS: Medline was searched via PubMed (1966 to May 2008) for anti-MCV and related terms, arthritis and arthropathies. Studies with anti-MCV, arthritis/arthropathy, and primary data on diagnosis and/or prognosis were included. Their methodological quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) instrument for diagnostic studies and the modified Hayden list for prognostic studies. RESULTS: Of 14 eligible studies, 11 included diagnostic data and 3 included prognostic data. No study evaluated anti-MCV as an added diagnostic test to the already available anti-cyclic citrullinated peptide (CCP) and rheumatoid factor serology. One study included the optimal patient spectrum resulting in a sensitivity of 0.59 and specificity of 0.98. A total of 10 diagnostic case-control studies using the same anti-MCV kit showed a sensitivity of 0.64-0.84 and a specificity of 0.79-0.96. This almost equalled the performance of anti-CCP in the same studies. The prognostic evaluation of anti-MCV was limited by differences in study methodology, outcome and statistical modelling. Individual studies showed moderate associations for anti-MCV and radiological progression with the strength of the association comparable to that of anti-CCP. CONCLUSIONS: Study heterogeneity, choice of study population and methodological limitations limited overall conclusions about the true diagnostic and prognostic test performance of anti-MCV. Evidence from the diagnostic case-control studies suggests that anti MCV may be used as an alternative for anti-CCP. PMID- 19289383 TI - Increased expression of the endothelin system in arterial lesions from patients with giant-cell arteritis: association between elevated plasma endothelin levels and the development of ischaemic events. AB - OBJECTIVE: Approximately 15-20% of patients with giant-cell arteritis (GCA) develop ischaemic complications often preceded by transient ischaemia. The expression of the endothelin (ET) system in GCA lesions was investigated to assess its relationship with the development of ischaemic complications. METHODS: Plasma ET-1 was quantified by immunoassay in 61 patients with biopsy-confirmed GCA and 16 healthy donors. ET-1, endothelin-converting enzyme (ECE-1) and endothelin receptor (ET(A)R and ET(B)R) messenger RNA were measured by real-time quantitative reverse transcriptase-PCR in temporal arteries from 35 of these patients and 19 control arteries. Proteins were measured by immunoassay and Western blot. RESULTS: ET-1 concentration was increased at the protein level in temporal artery samples from GCA patients compared with controls (0.98 (SEM 0.32) vs 0.28 (SEM 0.098) fmol/mg, p = 0.028). ECE-1, ET(A)R and ET(B)R/actin ratios (Western blot) were also significantly higher in GCA patients. Intriguingly, mRNA expression of ET-1, ECE-1 and both receptors was significantly reduced in GCA lesions compared with control arteries. When investigating mechanisms underlying these results, platelet-derived growth factor and IL-1beta, present in GCA lesions, were found to downregulate ET-1 mRNA in cultured human temporal artery derived smooth muscle cells. Glucocorticoid treatment for 8 days did not result in significantly decreased endothelin tissue concentration (0.87 (SEM 0.2) vs 0.52 (SEM 0.08); p = 0.6). Plasma endothelin concentrations were higher in patients with ischaemic complications (1.049 (SEM 0.48) vs 1.205 (SEM 0.63) pg/ml, p = 0.032). CONCLUSIONS: The endothelin system is increased at the protein level in GCA lesions creating a microenvironment prone to the development of ischaemic complications. Recovery induced by glucocorticoids is delayed, indicating persistent exposure to endothelin during initial treatment. PMID- 19289384 TI - National cohort study of reproductive risk factors for rheumatoid arthritis in Denmark: a role for hyperemesis, gestational hypertension and pre-eclampsia? AB - OBJECTIVES: While reproductive factors might plausibly be involved in the aetiology of rheumatoid arthritis (RA), the female predominance remains unexplained. A study was undertaken to address the possible impact of live births, pregnancy losses and pregnancy complications on the subsequent risk of RA in a nationwide cohort study. METHODS: National register data were used to link reproductive histories and later RA hospitalisations in a cohort of 4.4 million Danes. As a measure of relative risk associated with different reproductive histories, ratios of first inpatient RA hospitalisation rates (RRs) were used with 95% confidence intervals (CIs) obtained by Poisson regression analysis. RESULTS: Overall, 7017 women and 3041 men were admitted to hospital with RA in 1977-2004 (88.8 million person-years). The risk of RA was inversely associated with age at birth of first child in both women and men (p for trend <0.001). Overall, nulliparity and a history of pregnancy loss were not associated with RA risk but, compared with one-child mothers, women with two (RR 0.84; 95% CI 0.78 to 0.90) or three (RR 0.83; 95% CI 0.77 to 0.91) children were at reduced risk. The risk of RA was increased in women with a history of hyperemesis (RR 1.70; 95% CI 1.06 to 2.54), gestational hypertension (RR 1.49; 95% CI 1.06 to 2.02) or pre eclampsia (RR 1.42; 95% CI 1.08 to 1.84). CONCLUSIONS: One-child mothers and young parents are at increased risk of RA later in life, possibly due to socioeconomic factors. The novel finding of a significantly increased risk of RA in women whose pregnancies were complicated by hyperemesis, gestational hypertension or pre-eclampsia might reflect reduced immune adaptability to pregnancy in women disposed to RA or a role of fetal microchimerism in the aetiology of RA. PMID- 19289385 TI - Accuracy of a new partial coherence interferometry analyser for biometric measurements. AB - AIMS: Precise biometry is an essential preoperative measurement for refractive surgery as well as cataract surgery. A new device based on partial coherence interferometry technology was tested and evaluated for accuracy of measurements. METHODS: In a prospective study 200 eyes of 100 healthy phakic volunteers were examined with a functional prototype of the new ALLEGRO BioGraph (Wavelight AG)/LENSTAR LS 900 (Haag Streit AG) biometer and with the IOLMaster V.5 (Carl Zeiss Meditec AG). As recommended by the manufacturers, repeated measurements were performed with both devices and the results compared using Spearman correlation calculations (WinSTAT). RESULTS: Spearman correlation showed high correlations for axial length and keratometry measurements between the two devices tested. Anterior chamber depth, however, had a lower correlation between the two biometry devices. In addition, the mean values of the anterior chamber depth differed (IOLMaster 3.48 (SD 0.42) mm versus BioGraph/LENSTAR 3.64 (SD 0.26) mm); however, this difference was not statistically different (p>0.05, t test). CONCLUSION: The new biometer provided results that correlated very well with those of the IOLMaster. The ALLEGRO BioGraph/LENSTAR LS 900 is a precise device containing additional features that will be helpful tools for any cataract or refractive surgeon. PMID- 19289386 TI - Vascular protection in peripheral artery disease: systematic review and modelling study. AB - AIMS: To ascertain the effectiveness of medical therapy for reducing risk in peripheral artery disease (PAD) and to model the potential impact of combining multiple efficacious approaches. METHODS AND RESULTS: 17 electronic databases, reference lists of primary studies, clinical practice guidelines, review articles, trial registries and conference proceedings from cardiology, vascular surgery and atherosclerosis meetings were screened. Eligible studies were randomized trials or meta-analyses of randomized trials of medical therapy for PAD which reported major cardiovascular events (myocardial infarction, stroke and cardiovascular death). Baseline event rates for modelling analyses were derived from published natural history cohorts. Overall, three strategies had persuasive evidence for reducing risk in PAD: antiplatelet agents (pooled RRR 26%, 95% CI 10 to 42), statins (pooled RRR 26%, 95% CI 18 to 33) and angiotensin-converting enzyme inhibitors (individual trial RRR 25%, 95% CI 8 to 39). The estimated cumulative relative risk reduction for all three strategies was 59% (CI 32 to 76). Given a 5-year major cardiovascular event rate of 25%, the corresponding absolute risk reduction and number needed to treat to prevent one event were 15% (CI 8 to 19) and 7 (CI 5 to 12), respectively. Population level analyses suggest that increased uptake of these modalities could prevent more than 200 000 events in patients with PAD each year. CONCLUSION: The use of multiple efficacious strategies has the potential to substantially reduce the cardiovascular burden of PAD. However, these data should be regarded as hypothetical, since they are based on mathematical modelling rather than factorial randomized trials. PMID- 19289387 TI - Medium-term effects of Italian smoke-free legislation: findings from four annual population-based surveys. AB - BACKGROUND: Italy was the first large country to ban smoking in all indoor public places, including restaurants and bars. The aim of this study was to quantify, 3 years after the law came into force, the effects of the smoking ban in terms of observance of the legislation and change of habits. METHODS: Data were considered from four representative surveys on smoking, conducted between 2005 and 2008 on a total of 12 245 individuals (5906 men and 6339 women) aged 15 years or over. RESULTS: In 2008, more than 80% of Italians (more than 90% in northern Italy) had the perception that the smoking ban was respected in bars/cafes and restaurants, despite a slight reduction since 2005. In all the surveys combined, 75% of the Italian population reported that the smoking ban was respected in workplaces. Overall, approximately 10% of Italians reported that, after the implementation of the tobacco regulation, they went to bars/cafes and restaurants more frequently, and approximately 7% less frequently, than before. CONCLUSION: The study shows that in Italy the smoke-free legislation did not affect the business of restaurants and bars, and remains widely respected 3 years after the law came into force. PMID- 19289388 TI - Unemployment and mortality--a longitudinal prospective study on selection and causation in 49321 Swedish middle-aged men. AB - BACKGROUND: Unemployment is associated with increased risk of mortality. It is, however, not clear to what extent this is causal, or whether other risk factors remain uncontrolled for. The aim of this study was to investigate the association between unemployment and all-cause and cause-specific mortality, adjusting for indicators of mental disorder, behavioural risk factors and social factors over the life course. METHODS: This study was based on a cohort of 49321 Swedish males, born 1949/51, tested for compulsory military conscription in 1969/70. Data on employment/unemployment 1990-4 was based on information from the Longitudinal Register of Education and Labour Market Statistics. Information on childhood circumstances was drawn from National Population and Housing Census 1960. Information on psychiatric diagnosis and behavioral risk factors was collected at conscription testing in 1969/70. Data on mortality and hospitalisation 1973-2004 were collected in national registers. RESULTS: An increased risk of mortality 1995-2003 was found among individuals who experienced 90 days or more of unemployment during 1992-4 compared with those still employed (all-cause mortality HR 1.91, 95% CI 1.58 to 2.31. Adjustment for risk factors measured along the life course considerably lowered the relative risk (all cause mortality HR 1.30, 95% CI 1.06 to 1.58). Statistically significant increased relative risk was found during the first 4 years of follow up (all-cause mortality, adjusted HR 1.57, 95% CI 1.13 to 2.18, but not the following 4 years (all cause mortality, adjusted HR 1.17, 95% CI 0.91 to 1.50). CONCLUSION: The results suggest that a substantial part of the increased relative risk of mortality associated with unemployment may be attributable to confounding by individual risk factors. PMID- 19289389 TI - Effects of dietary fibre intake on risk factors for cardiovascular disease in subjects at high risk. AB - BACKGROUND: Epidemiological studies and feeding trials with supplements suggest that fibre intake is associated with a reduction in cardiovascular risk. However, the effects of changes in dietary fibre on risk factor levels have not been evaluated in free-living individuals. Thus, the effects of changes in dietary fibre intake on cardiovascular risk factors were assessed over 3 months in free living high-risk subjects. METHODS: 772 high-risk subjects (age 69+/-5 years) were assigned to a low-fat diet or two Mediterranean-style diets. All participants received behavioural and nutritional education, including recommendations for increasing the consumption of vegetables, fruits, and legumes. Changes in food and nutrient intake, body weight, blood pressure, lipid profiles, glucose control and inflammatory markers were evaluated. RESULTS: Most participants increased consumption of vegetable products, but the increase in dietary fibre exhibited wide between-subject variability (6-65 g/day). Body weight, waist circumference, and mean systolic and diastolic blood pressure decreased across quintiles of fibre intake (p<0.005; all). Reductions in fasting glucose and total cholesterol levels, and increments in HDL cholesterol were highest among participants in the upper 20% of fibre intake (p = 0.04 and 0.02 respectively). Plasma concentrations of C-reactive protein, but not those of inflammatory cytokines, decreased in parallel with increasing dietary fibre (p = 0.04). Significant reductions in LDL cholesterol were observed only among participants with the greatest increases in soluble fibre intake (p = 0.04). CONCLUSIONS: Increasing dietary fibre intake with natural foods is associated with reductions in classical and novel cardiovascular risk factors in a high-risk cohort. PMID- 19289390 TI - Rhinitis associated with pesticide exposure among commercial pesticide applicators in the Agricultural Health Study. AB - OBJECTIVES: Rhinitis is common, but the risk factors are not well described. To investigate the association between current rhinitis and pesticide use, we used data from 2245 Iowa commercial pesticide applicators in the Agricultural Health Study. METHODS: Using logistic regression models adjusted for age, education and growing up on a farm, we evaluated the association between current rhinitis and 34 pesticides used in the past year. RESULTS: 74% of commercial pesticide applicators reported at least one episode of rhinitis in the past year (current rhinitis). Five pesticides used in the past year were significantly positively associated with current rhinitis: the herbicides 2,4-D, glyphosate and petroleum oil, the insecticide diazinon and the fungicide benomyl. The association for 2,4 D and glyphosate was limited to individuals who used both in the past year (OR 1.42, 95% CI 1.14 to 1.77). Both petroleum oil and diazinon showed consistent evidence of an association with rhinitis, based on both current use and exposure response models. We saw no evidence of confounding by common agricultural rhinitis triggers such as handling grain or hay. CONCLUSIONS: Exposure to pesticides may increase the risk of rhinitis. PMID- 19289391 TI - Can loud noise cause acoustic neuroma? Analysis of the INTERPHONE study in France. AB - OBJECTIVES: To investigate possible associations between risk of acoustic neuroma and exposure to loud noise in leisure and occupational settings. METHODS: A case control study was conducted in France within the international INTERPHONE study. The cases were the 108 subjects diagnosed with acoustic neuroma between 1 June 2000 and 31 August 2003. Two controls per case were selected from the electoral rolls and individually matched for gender, age (5 years) and area (local authority district) of residence at the time of the case diagnosis. Multivariate analyses were conducted using conditional logistic regression. Adjustment was made for socioeconomic status. RESULTS: Acoustic neuroma was found to be associated with loud noise exposure (odds ratio (OR) = 2.55; 95% CI 1.35 to 4.82), both in leisure settings, particularly when listening to loud music (OR = 3.88; 95% CI 1.48 to 10.17) and at work (OR = 2.26; 95% CI 1.08 to 4.72). This risk increased with exposure duration (>6 years' leisure exposure: OR = 3.15; 95% CI 1.07 to 9.24). Risk varied according to the type of noise (continuous or explosive vs intermittent). CONCLUSION: The present results agree with other recent reports implicating loud noise in the risk of acoustic neuroma. PMID- 19289392 TI - Do mutations of the Pendred syndrome gene, SLC26A4, confer resistance to asthma and hypertension? AB - BACKGROUND AND AIMS: Mutations of SLC26A4 cause Pendred syndrome, an autosomal recessive disorder comprising goitre and deafness with enlarged vestibular aqueducts (EVA). Recent studies in mouse models implicate Slc26a4 in the pathogenesis of asthma and hypertension. We hypothesise that asthma and hypertension are less prevalent among humans with SLC26A4 mutations. METHODS: We reviewed medical histories and SLC26A4 genotypes for 80 individuals with EVA and 130 of their unaffected family members enrolled in a study of EVA. We used Fisher's exact test to compare the prevalence of asthma and hypertension among groups of subjects with zero, one, or two mutant alleles of SLC26A4. RESULTS: Although none of the 21 subjects with two mutant alleles of SLC26A4 had asthma or hypertension, there were no statistically significant differences in the prevalence of asthma or hypertension among subjects with zero, one, or two mutant alleles. CONCLUSION: There might be a protective effect of SLC26A4 mutations for asthma and hypertension but our study is statistically underpowered to detect this effect. Study sizes of at least 1125 and 504 individuals will be needed for 80% power to detect an effect at alpha = 0.05 for asthma and hypertension, respectively. Our hypothesis merits a larger study since it has implications for potential strategies to treat hearing loss by manipulating SLC26A4 expression or function. PMID- 19289393 TI - Microdeletion 15q13.3: a locus with incomplete penetrance for autism, mental retardation, and psychiatric disorders. AB - BACKGROUND: Microdeletions within chromosome 15q13.3 are associated both with a recently recognised syndrome of mental retardation, seizures, and dysmorphic features, and with schizophrenia. METHODS AND RESULTS: Based on routine diagnostic testing of approximately 8200 samples using array comparative genomic hybridisation, we identified 20 individuals (14 children and six parents in 12 families) with microdeletions of 15q13.3. Phenotypes in the children included developmental delay, mental retardation, or borderline IQ in most and autistic spectrum disorder (6/14), speech delay, aggressiveness, attention deficit hyperactivity disorder, and other behavioural problems. Both parents were available in seven families, and the deletion was de novo in one, inherited from an apparently normal parent in four, and inherited from a parent with learning disability and bipolar disorder in two families. Of the 14 children, six in five families were adopted, and DNA was available for only one of these 10 biological parents; the deletion was very likely inherited for one of these families with two affected children. Among the unavailable parents, two mothers were described as having mental retardation, another mother as having "mental illness", and one father as having schizophrenia. We hypothesise that some of the unavailable parents have the deletion. CONCLUSIONS: The occurrence of increased adoption, frequent autism, bipolar disorder, and lack of penetrance are noteworthy findings in individuals with deletion 15q13.3. A high rate of adoption may be related to the presence of the deletion in biological parents. Unconfirmed histories of antisocial behaviours in unavailable biological parents raise the concern that future research may show that deletion 15q13.3 is associated with such behaviours. PMID- 19289394 TI - Recessive primary congenital lymphoedema caused by a VEGFR3 mutation. AB - BACKGROUND: Heterozygous mutations in VEGFR3 have been identified in some familial cases with dominantly inherited primary congenital lymphoedema, known as Nonne-Milroy disease. Recessive cases of primary lymphoedema with a genetic cause are not known, except for two families with syndromic hypotrichosis-lymphoedema telangiectasia, with a SOX18 mutation. METHODS AND RESULTS: In this study, we present the first case of isolated primary congenital lymphoedema with recessive inheritance, caused by a homozygous mutation in VEGFR3. The novel mutation is a transition from alanine-to-threonine in amino acid 855, located in the ATP binding domain of the VEGFR3 receptor. Assessment of receptor function showed impaired ligand induced internalisation and ERK1/2 activity. Moreover, receptor phosphorylation was reduced, although less so than for a kinase-dead VEGFR3 mutation, which causes Nonne-Milroy disease. CONCLUSION: A hypomorphic VEGFR3 mutation, with moderate effect on receptor function, in a homozygous state can result in insufficient lymphatic functioning. Thus, in addition to Nonne-Milroy disease with dominant inheritance, VEGFR3 alterations can cause isolated recessive primary congenital lymphoedema. These data expand our understanding of the aetiology of congenital lymphoedema and suggest that large scale screening of VEGFR3 in all primary lymphoedema patients is necessary. PMID- 19289395 TI - HER-2 as a target for breast cancer therapy. PMID- 19289397 TI - Morphologic features of biocompatibility and neoangiogenesis onto a biodegradable tracheal prosthesis in an animal model. AB - We evaluated a newly designed bioresorbable polymer (Degrapol) tracheal prosthesis in an in-vivo angiogenesis-inducing animal model focusing on the specific tissue reaction, the neo-angiogenesis and also the eventual cathepsin B role during the polymer degradation. Fifteen rabbits were divided into three groups (2, 6 and 8 weeks) and our tube-shaped porous prosthesis was implanted using the common carotid artery and the internal jugular vein as vascular pedicle. Optical and electron microscopy, immunohistochemistry and immunocytochemistry were performed at the end of each period, showing cells and fibrils, in direct contact with the Degrapol scaffold, strongly increased with time. Blood vessel neoformation was visible with CD31 expression localized at the endothelial cells forming the neovascular walls. Over time many of them differentiate in muscle fibers as validated by the expression of alpha-smooth muscle actin (SMA). Few inflammatory cells, expressing CD14, were visible while most cells adopting a pronounced spreading phenotype showed a strong positivity for cathepsin B. We concluded that this bioresorbable polymer provided a good substrate for fibrous tissue deposition with an excellent degree of neo angiogenesis. Also, cathepsin B seems to contribute to the polymer degradation and particularly to neovascularization by stimulating capillary-like tubular structures and cell proliferation. PMID- 19289398 TI - External iliac artery pseudoaneurysm complicating renal transplantation. AB - OBJECTIVES: To assess the etiology, management and outcome of iliac pseudoaneurysms following renal transplantation. METHODS: Eleven patients who underwent repair between 1982 and 2007 were identified. Five (Group 1) presented pseudoaneurysm at the anastomosis of the donor renal and native iliac arteries, and six (Group 2) presented iliac pseudoaneurysm following transplant nephrectomy. Intraoperative cultures and immunohistochemical examinations were obtained from all surgical cases to determine the existence of a relationship between infection or transplant rejection and pseudoaneurysm formation. RESULTS: Endovascular repair (EVR) was used to treat three patients, while eight patients underwent open repair (OR). Transplant nephrectomy was needed in all cases but one after anastomotic pseudoaneurysm repair. After pseudoaneurysm excision, arterial reconstruction was performed in all cases, with a limb salvage rate of 100%. At 30 days, no patients died in the EVR subgroup. In the OR subgroup, one patient died of sepsis (12.5%). Cultures taken from the pseudoaneurysm wall and content grew Candida albicans and E. coli in two febrile patients. Pathologic evaluation of donor renal arteries revealed evidence of chronic rejection in three patients (60%) in Group 1, and in two (33.3%) in Group 2. No patients in either Group presented late infection, failure of vascular reconstruction nor pseudoaneurysm recurrence. The follow-up ranges from 20 to 89 months. CONCLUSIONS: The etiology of pseudoaneurysms in this location is multifactorial, however, an association with chronic rejection must be considered. Though rare, the development of pseudoaneurysms at the donor renal-external iliac artery anastomosis results in high rates of transplant nephrectomy. Less invasive endovascular techniques offer a new therapeutic option in this challenging scenario notwithstanding the fact that they require further validation. PMID- 19289399 TI - Protective effect of sivelestat sodium hydrate (ONO-5046) on ischemic spinal cord injury. AB - Prevention of paraplegia remains an important issue in repair of descending thoracic and thoracoabdominal aneurysms. Therefore, we investigated the protective effect of sivelestat sodium hydrate (ONO-5046) on ischemia-induced spinal cord damage in a rabbit model. Twenty New Zealand white rabbits were divided into two equal groups; ONO-5046 (1.6 mg/kg)+isotonic NaCl (30 ml) was administered selectively to the spinal cord via the lumbar arteries for the first 3 min during 30 min of infra-renal aorta clamping in the experimental group (group E), whereas NaCl was given alone in the control group (group C). Motor function of the lower limbs was assessed two days later by Tarlov criteria. The number of intact motor neurons in the anterior segment of the cord (L5 level) was counted after hematoxylin-eosin staining and the number of apoptotic motor neurons after TUNEL staining. Motor function of the lower limbs in group E was significantly better (P=0.003) than that in group C. The number of intact motor neurons was greater and of apoptotic motor neurons was less in group E than C. Selective infusion of sivelestat sodium hydrate directly into the spinal cord via the lumbar arteries significantly attenuated functional and morphological ischemia-induced spinal cord injury. PMID- 19289400 TI - Effective dose conversion factors in paediatric interventional cardiology. AB - Conversion factors for effective dose (CF(E) = effective dose/dose-area product (mSv (Gy cm2)(-1)) in paediatric interventional cardiology were estimated retrospectively for 249 patients using the dose-area product (DAP), irradiation geometry, exposure parameters and tissue-weighting factors (TWFs) from the International Commission on Radiological Protection (ICRP) 60. Two methods for estimating the conversion factors, which differed in the description of the irradiation geometry, were evaluated. The effective doses obtained with the two methods were almost identical. The results showed that irradiation geometry had no significant impact on the CF(E), and a single factor was defined for both diagnostic and interventional examinations. In addition, the effect of the new tissue-weighting factor for breast tissue (TWF(b)) given in ICRP 103 on the effective dose was assessed. The CF(E) was 3.7+/-0.2 mSv (Gy cm2)(-1) (neonate), 1.9+/-0.2 mSv (Gy cm2)(-1) (1 year), 1.0+/-0.1 mSv (Gy cm2)(-1) (5 years), 0.6+/ 0.1 mSv (Gy cm2)(-1) (10 years) and 0.4+/-0.1 mSv (Gy cm2)(-1) (15 years). Applying these CFs to the individual DAP values of each patient yielded mean effective doses of 13.0 mSv (neonate), 8.6 mSv (1 year), 6.4 mSv (5 years), 8.6 mSv (10 years) and 12.7 mSv (15 years). The maximum estimated skin dose (15 patients) did not exceed 60 mGy. With the new ICRP value for TWF(b), increases in the CFs in the order of 10-30%, and in the effective dose of 10-20%, were indicated. The results indicated that the effective dose in paediatric interventional cardiology is of much greater concern than the skin dose. Furthermore, age-dependent CF(E) values are required so as not to underestimate the doses to very young patients. PMID- 19289401 TI - Simple methods to reduce patient dose in a Varian cone beam CT system for delivery verification in pelvic radiotherapy. AB - Cone-beam computed tomography (CBCT) is a three-dimensional imaging modality that has recently become available on linear accelerators for radiotherapy patient position verification. It was the aim of the present study to implement simple strategies for reduction of the dose delivered in a commercial CBCT system. The dose delivered in a CBCT procedure (Varian, half-fan acquisition, 650 projections, 125 kVp) was assessed using a cylindrical Perspex phantom (diameter, 32 cm) with a calibrated Farmer type ionisation chamber. A copper filter (thickness, 0.15 mm) was introduced increasing the half value layer of the beam from 5.5 mm Al to 8 mm Al. Image quality and noise were assessed using an image quality phantom (CatPhan) while the exposure settings per projection were varied from 25 ms/80 mA to 2 ms/2 mA per projection. Using the copper filter reduced the dose to the phantom from approximately 45 mGy to 30 mGy at standard settings (centre/periphery weighting 1/3 to 2/3). Multiple CBCT images were acquired for six patients with pelvic malignancies to compare CBCTs with and without a copper filter. Although the reconstructed image is somewhat noisier with the filter, it features similar contrast in the centre of the patient and was often preferred by the radiation oncologist because of greater image uniformity. The X-ray shutters were adjusted to the minimum size required to obtain the desired image volume for a given patient diameter. The simple methods described here reduce the effective dose to patients undergoing daily CBCT and are easy to implement, and initial evidence suggests that they do not affect the ability to identify soft tissue for the purpose of treatment verification. PMID- 19289402 TI - Thorax set-up verification with multiple oblique treatment portal images. AB - Patient set-up in external beam radiotherapy to the thorax is routinely checked by matching anterior and lateral ports to simulator or digitally reconstructed radiograph (DRR) reference images. We report a method to derive bed shifts from matching multiple oblique treatment ports, exploiting data redundancy for match consistency checking. Portal images were acquired for 14 thorax patients on the first 3 treatment days and matched to DRRs. As per clinical practice, anterior and lateral portal images were matched and checked. In addition, treatment ports were acquired and matched and field placement errors calculated from equations generating lateral and ventrodorsal bed shifts from any pair of treatment fields. We compared bed moves obtained from clinical imaging and from oblique treatment fields. Inconsistencies larger than 3 mm triggered a further independent check using new reference anatomy. With clinical image matching, set-up errors had a 95% confidence interval of +/-8 mm ventrodorsal and +/-6 mm in the lateral direction. With independent oblique field re-evaluation, the confidence intervals were reduced to +/-3 mm and +/-2 mm, respectively. Matching using the oblique fields detected set-up errors greater than 5 mm in 16 out of 90 matches. In the clinical environment, match errors greater than 5 mm in the thorax area can remain undetected using anterior and lateral fields only. Consequently, necessary bed shifts are not made or made in error. We developed a technique that uses portal images from multiple oblique treatment fields and exploits data redundancy for internal match consistency checking. PMID- 19289403 TI - Genetic interaction between the m-AAA protease isoenzymes reveals novel roles in cerebellar degeneration. AB - The mitochondrial m-AAA protease has a crucial role in axonal development and maintenance. Human mitochondria possess two m-AAA protease isoenzymes: a hetero oligomeric complex, composed of paraplegin and AFG3L2 (Afg3 like 2), and a homo oligomeric AFG3L2 complex. Loss of function of paraplegin (encoded by the SPG7 gene) causes hereditary spastic paraplegia, a disease characterized by retrograde degeneration of cortical motor axons. Spg7(-/-) mice show a late-onset degeneration of long spinal and peripheral axons with accumulation of abnormal mitochondria. In contrast, Afg3l2(Emv66/Emv66) mutant mice, lacking the AFG3L2 protein, are affected by a severe neuromuscular phenotype, due to defects in motor axon development. The role of the homo-oligomeric m-AAA protease and the extent of cooperation and redundancy between the two isoenzymes in adult neurons are still unclear. Here we report an early-onset severe neurological phenotype in Spg7(-/-) Afg3l2(Emv66/+) mice, characterized by loss of balance, tremor and ataxia. Spg7(-/-) Afg3l2(Emv66/+) mice display acceleration and worsening of the axonopathy observed in paraplegin-deficient mice. In addition, they show prominent cerebellar degeneration with loss of Purkinje cells and parallel fibers, and reactive astrogliosis. Mitochondria from affected tissues are prone to lose mt-DNA and have unstable respiratory complexes. At late stages, neurons contain structural abnormal mitochondria defective in COX-SDH reaction. Our data demonstrate genetic interaction between the m-AAA isoenzymes and suggest that different neuronal populations have variable thresholds of susceptibility to reduced levels of the m-AAA protease. Moreover, they implicate impaired mitochondrial proteolysis as a novel pathway in cerebellar degeneration. PMID- 19289404 TI - Short 42 degrees C heat shock induces phosphorylation and degradation of Cdc25A which depends on p38MAPK, Chk2 and 14.3.3. AB - The effects of heat shock (HS; 42 degrees C) on the cell cycle and underlying molecular mechanisms are astonishingly unexplored. Here, we show that HS caused rapid Cdc25A degradation and a reduction of cell cycle progression. Cdc25A degradation depended on Ser75-Cdc25A phosphorylation caused by p38MAPK and Chk2, which phosphorylated Ser177-Cdc25A that is specific for 14.3.3 binding. Upon HS, Cdc25A rapidly co-localized with 14.3.3 in the perinuclear space that was accompanied with a decrease of nuclear Cdc25A protein levels. Consistently, a 14.3.3 binding-deficient Cdc25A double mutant (Ser177/Ala-Tyr507/Ala) was not degraded in response to HS and there was no evidence for an increased co localization of Cdc25A with 14.3.3 in the cytosol. Therefore, upon HS, p38, Chk2 and 14.3.3 were antagonists of Cdc25A stability. On the other hand, Cdc25A was protected by Hsp90 in HEK293 cells because the specific inhibition of Hsp90 with Geldanamycin caused Cdc25A degradation in HEK293 implicating that Cdc25A is an Hsp90 client. Specific inhibition of Hsp90 together with HS caused and accelerated degradation of Cdc25A and was highly cytotoxic. The results presented here show for the first time that Cdc25A is degraded by moderate heat shock and protected by Hsp90. We describe the mechanisms explaining HS-induced cell cycle retardation and provide a rationale for a targeted hyperthermia cancer therapy. PMID- 19289409 TI - Surgeons have insufficient experience to operate on children, report warns. PMID- 19289410 TI - Health service should develop expertise in child protection, review urges. PMID- 19289411 TI - Reducing greenhouse emissions could cut global disease burden by 25%. PMID- 19289412 TI - Treatment of intermittent claudication. PMID- 19289413 TI - Management of rheumatoid arthritis: summary of NICE guidance. PMID- 19289414 TI - Zimbabwe's health challenges. PMID- 19289415 TI - Saving the planet as well as lives. PMID- 19289416 TI - L-FABP directly interacts with PPARalpha in cultured primary hepatocytes. AB - Although studies with liver type fatty acid binding protein (L-FABP) gene ablated mice demonstrate a physiological role for L-FABP in hepatic fatty acid metabolism, little is known about the mechanisms whereby L-FABP elicits these effects. Studies indicate that L-FABP may function to shuttle lipids to the nucleus, thereby increasing the availability of ligands of nuclear receptors, such as peroxisome proliferator-activated receptor-alpha (PPARalpha). The data herein suggest that such mechanisms involve direct interaction of L-FABP with PPARalpha. L-FABP was shown to directly interact with PPARalpha in vitro through co-immunoprecipitation (co-IP) of pure proteins, altered circular dichroic (CD) spectra, and altered fluorescence spectra. In vitro fluorescence resonance energy transfer (FRET) between Cy3-labeled PPARalpha and Cy5-labeled L-FABP proteins showed that these proteins bound with high affinity (Kd approximately 156 nM) and in close proximity (intermolecular distance of 52A). This interaction was further substantiated by co-IP of both proteins from liver homogenates of wild-type mice. Moreover, double immunogold electron microscopy and FRET confocal microscopy of cultured primary hepatocytes showed that L-FABP was in close proximity to PPARalpha (intermolecular distance 40-49A) in vivo. Taken together, these studies were consistent with L-FABP regulating PPARalpha transcriptional activity in hepatocytes through direct interaction with PPARalpha. Our in vitro and imaging experiments demonstrate high affinity, structural molecular interaction of L-FABP with PPARalpha and suggest a functional role for L-FABP interaction with PPARalpha in long chain fatty acid (LCFA) metabolism. PMID- 19289417 TI - Overexpression of sterol carrier protein-2 differentially alters hepatic cholesterol accumulation in cholesterol-fed mice. AB - Although in vitro studies suggest a role for sterol carrier protein-2 (SCP-2) in cholesterol trafficking and metabolism, the physiological significance of these observations remains unclear. This issue was addressed by examining the response of mice overexpressing physiologically relevant levels of SCP-2 to a cholesterol rich diet. While neither SCP-2 overexpression nor cholesterol-rich diet altered food consumption, increased weight gain, hepatic lipid, and bile acid accumulation were observed in wild-type mice fed the cholesterol-rich diet. SCP-2 overexpression further exacerbated hepatic lipid accumulation in cholesterol-fed females (cholesterol/cholesteryl esters) and males (cholesterol/cholesteryl esters and triacyglycerol). Primarily in female mice, hepatic cholesterol accumulation induced by SCP-2 overexpression was associated with increased levels of LDL-receptor, HDL-receptor scavenger receptor-B1 (SR-B1) (as well as PDZK1 and/or membrane-associated protein 17 kDa), SCP-2, liver fatty acid binding protein (L-FABP), and 3alpha-hydroxysteroid dehydrogenase, without alteration of other proteins involved in cholesterol uptake (caveolin), esterification (ACAT2), efflux (ATP binding cassette A-1 receptor, ABCG5/8, and apolipoprotein A1), or oxidation/transport of bile salts (cholesterol 7alpha-hydroxylase, sterol 27alpha hydroxylase, Na(+)/taurocholate cotransporter, Oatp1a1, and Oatp1a4). The effects of SCP-2 overexpression and cholesterol-rich diet was downregulation of proteins involved in cholesterol transport (L-FABP and SR-B1), cholesterol synthesis (related to sterol regulatory element binding protein 2 and HMG-CoA reductase), and bile acid oxidation/transport (via Oapt1a1, Oatp1a4, and SCP-x). Levels of serum and hepatic bile acids were decreased in cholesterol-fed SCP-2 overexpression mice, especially in females, while the total bile acid pool was minimally affected. Taken together, these findings support an important role for SCP-2 in hepatic cholesterol homeostasis. PMID- 19289418 TI - PET imaging of prostate cancer xenografts with a highly specific antibody against the prostate-specific membrane antigen. AB - Prostate-specific membrane antigen (PSMA), a transmembrane glycoprotein, is highly expressed by virtually all prostate cancers and is currently the focus of several diagnostic and therapeutic strategies. We have previously reported on the generation of several monoclonal antibodies (mAb) and antibody fragments that recognize and bind with high affinity to the extracellular domain of cell adherent PSMA. This article reports the in vivo behavior and tumor uptake of the radiolabeled anti-PSMA mAb 3/A12 and its potential as a tracer for PET. METHODS: The mAb 3/A12 was conjugated with the chelating agent 1,4,7,10 tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) and radiolabeled with (64)Cu. Severe combined immunodeficient mice bearing PSMA-positive C4-2 prostate carcinoma xenografts were used for small-animal PET imaging. Mice with PSMA negative DU 145 tumors served as controls. For PET studies, each animal received 20-30 microg of radiolabeled mAb corresponding to an activity of 7.6-11.5 MBq. Imaging was performed 3, 24, and 48 h after injection. After the last scan, the mice were sacrificed and tracer in vivo biodistribution was measured by gamma counting. RESULTS: Binding of the mAb 3/A12 on PSMA-expressing C4-2 cells was only minimally influenced by DOTA conjugation. The labeling efficiency using (64)Cu and DOTA-3/A12 was 95.3% +/- 0.3%. The specific activity after (64)Cu labeling was between 327 and 567 MBq/mg. After tracer injection, static small animal PET images of mice with PSMA-positive tumors revealed a tumor-to background ratio of 3.3 +/- 1.3 at 3 h, 7.8 +/- 1.4 at 24 h, and 9.6 +/- 2.7 at 48 h. In contrast, no significant tracer uptake occurred in the PSMA-negative DU 145 tumors. These results were confirmed by direct counting of tissues after the final imaging. CONCLUSION: Because of the high and specific uptake of (64)Cu labeled mAb 3/A12 in PSMA-positive tumors, this ligand represents an excellent candidate for prostate cancer imaging and potentially for radioimmunotherapy. PMID- 19289419 TI - 111In-LLP2A-DOTA Polyethylene Glycol-Targeting {alpha}4{beta}1 Integrin: Comparative Pharmacokinetics for Imaging and Therapy of Lymphoid Malignancies. AB - N-[[4-[[[(2-ethylphenyl)amino]carbonyl]amino]phenyl]acetyl]-N(epsilon)-6-[(2E)-1 oxo-3-(3-pyridinyl-2-propenyl)]-l-lysyl-l-2-aminohexanedioyl-(1-amino-1 cyclohexane)carboxamide (LLP2A) is a high-affinity, high-specificity peptidomimetic ligand (inhibitory concentration of 50% = 2 pM) that binds the activated alpha4beta1 integrin found on a variety of malignant lymphoid cell lines. To better determine whether this ligand holds promise for imaging and therapy in lymphoid malignancies, 6 LLP2A derivatives, as LLP2A-1,4,7,10 tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (LLP2A-DOTA) and LLP2A-DOTA polyethylene glycol (LLP2A-DOTA-PEG), were designed, synthesized, and radiolabeled with (111)In. Comparative pharmacokinetic studies in mice with Raji B-cell lymphoma xenografts were then complemented by small-animal PET of the lead molecular LLP2A format using (64)Cu-LLP2A-11-bis(carboxymethyl)-1,4,8,11 tetraazabicyclo[6.6.2]hexadecane ((64)Cu-LLP2A-CB-TE2A). METHODS: LLP2A-DOTA and LLP2A-CB-TE2A were prepared using solid-phase synthesis; LLP2A-DOTA-PEG(2,000), LLP2A-DOTA-PEG(5,000), LLP2A-DOTA-PEG(10,000), (LLP2A-DOTA)(2)PEG(10,000), and (LLP2A-DOTA)(4)PEG(10,000) were prepared by PEGylation. (111)In radiolabeling of DOTA and (64)Cu radiolabeling of CB-TE2A conjugates yielded 370-1,850 and 3,700 7,400 kBq/microg (10-50 and 100-200 microCi/microg), respectively. The pharmacokinetics of the six (111)In radioconjugates were studied in vivo using biodistribution data (4 and 24 h) and whole-body autoradiography (24 h) in mice with Raji tumor xenografts. (64)Cu-LLP2A-CB-TE2A was imaged (4 and 24 h) on a small-animal PET scanner in the same mouse model. RESULTS: The highest tumor uptake in pharmacokinetic studies was obtained with LLP2A-DOTA and (LLP2A DOTA)(4)-PEG(10,000). For (111)In-LLP2A-DOTA (1 nM) at 4 and 24 h after injection, ratios of tumor to blood and tumor to nontumor (normal) organ (T/NT) were 8 to 35:1 for all organs or tissue except the spleen, marrow, and kidney, which were between 2:1 and 1:1. Tetravalent (LLP2A-DOTA)(4)-PEG(10,000) (1.1 nM) had tumor uptake similar to the univalent LLP2A-DOTA but higher liver, marrow, and kidney uptake. The excellent T/NT of LLP2A was also demonstrated by small animal PET with (64)Cu-LLP2A-CB-TE2A at both 4 and 24 h after injection; obvious spleen targeting was apparent, but little kidney or liver activity was observed. CONCLUSION: Of the conjugates investigated, the univalent, non-PEGylated ligand (111)In-LLP2A-DOTA exhibited the best T/NT ratios and showed the greatest potential for imaging of alpha4beta1 in human lymphoma. Furthermore, this univalent non-PEGylated LLP2A format, as (64)Cu-LLP2A-CB-TE2A, demonstrated excellent tumor targeting by small-animal PET and warrants further investigation as an agent for the study of alpha4beta1 expression in human lymphoid malignancies. PMID- 19289420 TI - 18F-FDOPA PET and PET/CT accurately localize pheochromocytomas. AB - Successful treatment of pheochromocytoma requires accurate diagnosis and localization of tumors. Herein, we investigated the accuracy of PET using 3,4 dihydroxy-6-(18)F-fluoro-phenylalanine ((18)F-FDOPA), an amino acid transporter substrate, as an independent marker for detection of benign and malignant pheochromocytomas. METHODS: The study comprised 25 consecutive patients (9 men, 16 women) whose median age was 51 y (range, 25-68 y), with known or suspected pheochromocytoma. Eleven patients underwent standardized (18)F-FDOPA PET and 14 patients underwent (18)F-FDOPA PET/CT studies, with a median of 511 MBq of (18)F FDOPA (range, 206-625 MBq). Two readers, unaware of the reports of other imaging studies and clinical data, analyzed all scans visually and quantitatively (maximum standardized uptake value [SUVmax] and maximum transverse diameter). Histology and long-term clinical follow-up served as the gold standard. Correlation between SUVmax of tumors and biochemical markers was evaluated. SUVmax of the benign and malignant tumors was compared. RESULTS: Seventeen patients underwent surgery. Histology confirmed pheochromocytoma or paraganglioma in 11 cases (8 adrenal, including 2 malignant tumors, and 3 extraadrenal, including 1 malignant tumor). The diagnosis of pheochromocytoma was established by follow-up in 2 additional patients (1 adrenal and 1 unknown location) and ruled out in 6 patients. Visual analysis detected and localized pheochromocytoma in 11 of 13 patients without false-positive results (sensitivity, 84.6%; specificity, 100%; accuracy, 92%). These lesions had an SUVmax of 2.3-34.9 (median, 8.3). Evaluation of the false-negative cases revealed a 13 x 5 mm lesion with an SUVmax of 1.96 in 1 case; no lesion was localized in the second case using multiple additional modalities. Spearman nonparametric analysis did not show statistically significant correlation between SUVmax of the tumors and biochemical markers. The Mann-Whitney nonparametric test did not demonstrate a statistically significant difference between the SUVmax of (18)F-FDOPA in malignant and benign tumors. CONCLUSION: (18)F-FDOPA PET and PET/CT are highly sensitive and specific tools that can provide additional independent information for diagnosis and localization of benign and malignant pheochromocytomas. PMID- 19289422 TI - The role of radiotracer imaging in the diagnosis and management of patients with breast cancer: part 1--overview, detection, and staging. AB - Breast cancer is the most common non-skin type of cancer and the second leading cause of cancer mortality in women. Advances in diagnosis and treatment have led to declines in mortality, despite an increase in breast cancer incidence. An advancing array of both local and systemic therapy options has led to increasingly individualized treatment. Imaging plays a key role in detecting breast cancer and directing its therapy. This continuing education article, part 1 in a 2-part series, provides a comprehensive review of current and future radiotracer imaging methods applied to breast cancer, in the context of breast cancer management strategies and other nonnuclear imaging methods. Part 1 of the review provides an overview of clinical and biologic considerations in breast cancer and covers radionuclide imaging for detection and staging. Part 2 will cover radionuclide imaging of breast cancer response to therapy, other clinical indications for radionuclide breast cancer imaging, and future directions, including molecular imaging. PMID- 19289423 TI - Auger radiation-induced, antisense-mediated cytotoxicity of tumor cells using a 3 component streptavidin-delivery nanoparticle with 111In. AB - When antisense oligomers are intracellular, they migrate to and are retained in the nucleus of tumor cells and therefore may be used to carry Auger electron emitting radionuclides such as (111)In for effective tumor radiotherapy. METHODS: Our nanoparticle consists of streptavidin that links 3 biotinylated components: the antiHer2 antibody trastuzumab (to improve pharmacokinetics), the tat peptide (to improve cell membrane transport), and the (111)In-labeled antiRIalpha messenger RNA antisense morpholino (MORF) oligomer. RESULTS: As evidence of unimpaired function, tumor cell and nuclear accumulations were orders of magnitude higher after incubation with (99m)Tc-MORF/tat/trastuzumab than after incubation with free (99m)Tc-MORF and significantly higher with the antisense than with the sense MORF. In mice, tumor and normal-tissue accumulations of the (99m)Tc-MORF/tat/trastuzumab nanoparticle were comparable to those of free (99m)Tc-trastuzumab, confirming the improved pharmacokinetics due to the trastuzumab component. Although kidneys, liver, and other normal tissues also accumulated the nanoparticle, immunohistochemical evaluation of tissue sections in mice receiving the Cy3-MORF/tat/trastuzumab nanoparticle showed evidence of nuclear accumulation only in tumor tissue. In a dose escalation study, as measured by the surviving fraction, the nanoparticle significantly increased the kill of SK-BR-3 breast cancer Her2+/RIalpha+ cells, compared with all controls. CONCLUSION: Significant radiation-induced antisense-mediated cytotoxicity of tumor cells in vitro was achieved using an Auger electron-emitting antisense MORF oligomer administered as a member of a 3-component streptavidin-delivery nanoparticle. PMID- 19289424 TI - Prognostic value of interim 18F-FDG PET in patients with diffuse large B-Cell lymphoma: SUV-based assessment at 4 cycles of chemotherapy. AB - Interim (18)F-FDG PET (after 1-4 cycles of chemotherapy) may be useful for tailoring a risk-adapted therapeutic strategy in lymphoma. The purpose of this study was to investigate whether semiquantification of standardized uptake values (SUVs) may help to improve the prognostic value of (18)F-FDG PET, compared with visual analysis, after 4 cycles of chemotherapy. METHODS: In a previous report, we showed that a 65.7% reduction in maximal SUV (SUVmax) between baseline (PET0) and 2 cycles of chemotherapy (PET2) better predicted event-free survival in 92 prospective patients with diffuse large B-cell lymphoma, by reducing false positive interpretation of visual analysis. Eighty patients also underwent (18)F FDG PET after induction had been completed, at 4 cycles of chemotherapy (PET4). Images were interpreted visually (as negative or positive) and by computing the optimal percentage of SUVmax reduction between PET0 and PET4. Survival curves were estimated using Kaplan-Meier analysis and compared using the log-rank test. Median follow-up was 41 mo. RESULTS: With visual analysis, the 2-y estimate for event-free survival was 82% in the PET4-negative group, compared with 25% in the PET4-positive group (P < 0.0001, accuracy of predicting event-free survival, 81.3%). An optimal cutoff of 72.9% SUVmax reduction from PET0 to PET4 yielded a 2 y estimate for event-free survival of 79% in patients with reduction of more than 72.9%, versus 32% in those with reduction of 72.9% or less (P < 0.0001; accuracy of predicting event-free survival, 77.5%). CONCLUSION: Although SUV semiquantification helps reduce false-positive interim (18)F-FDG PET interpretations at 2 cycles, its performance is equivalent to visual analysis at 4 cycles, when most of the therapeutic effect has occurred upstream. This approach may be useful for objectively tailoring consolidation strategies. PMID- 19289425 TI - U-SPECT-II: An Ultra-High-Resolution Device for Molecular Small-Animal Imaging. AB - We present a new rodent SPECT system (U-SPECT-II) that enables molecular imaging of murine organs down to resolutions of less than half a millimeter and high resolution total-body imaging. METHODS: The U-SPECT-II is based on a triangular stationary detector set-up, an XYZ stage that moves the animal during scanning, and interchangeable cylindric collimators (each containing 75 pinhole apertures) for both mouse and rat imaging. A novel graphical user interface incorporating preselection of the field of view with the aid of optical images of the animal focuses the pinholes to the area of interest, thereby maximizing sensitivity for the task at hand. Images are obtained from list-mode data using statistical reconstruction that takes system blurring into account to increase resolution. RESULTS: For (99m)Tc, resolutions determined with capillary phantoms were smaller than 0.35 and 0.45 mm using the mouse collimator with 0.35- and 0.6-mm pinholes, respectively, and less than 0.8 mm using the rat collimator with 1.0-mm pinholes. Peak geometric sensitivity is 0.07% and 0.18% for the mouse collimator with 0.35- and 0.6-mm pinholes, respectively, and 0.09% for the rat collimator. Resolution with (111)In, compared with that with (99m)Tc, was barely degraded, and resolution with (125)I was degraded by about 10%, with some additional distortion. In vivo, kidney, tumor, and bone images illustrated that U-SPECT-II could be used for novel applications in the study of dynamic biologic systems and radiopharmaceuticals at the suborgan level. CONCLUSION: Images and movies obtained with U-SPECT-II provide high-resolution radiomolecule visualization in rodents. Discrimination of molecule concentrations between adjacent volumes of about 0.04 microL in mice and 0.5 microL in rats with U-SPECT-II is readily possible. PMID- 19289426 TI - Fibronectin stimulates endothelial cell 18F-FDG uptake through focal adhesion kinase-mediated phosphatidylinositol 3-kinase/Akt signaling. AB - There has been recent interest in the relationship between (18)F-FDG uptake and the angiogenic activity of endothelial cells (ECs). The angiogenic process is strongly dependent on the interaction of ECs with matrix fibronectin (FN), a key regulator of EC survival, migration, and proliferation. Therefore, we investigated how FN influences EC glucose uptake and elucidated the signaling pathways that mediate this effect. METHODS: Human umbilical vein ECs were allowed to adhere to FN-coated plates and were compared with control cells for (18)F-FDG uptake, membrane GLUT1 levels, and hexokinase activity. The roles of focal adhesion kinase (FAK), phosphatidylinositol 3-kinase (PI3K), and Akt were evaluated with Western blotting, small interfering RNA (siRNA), and specific inhibitors. RESULTS: FN adhesion significantly enhanced the protein-corrected (18)F-FDG uptake in HUVEC, to 2.1-, 2.7-, and 4.3-fold that in control cells by 2, 3, and 5 d, respectively. This effect was mediated by the upregulation of both membrane GLUT1 expression and hexokinase activity and was accompanied by FAK activation. Silencing of FAK signaling by siRNA completely abrogated both FN induced FAK phosphorylation and (18)F-FDG uptake. FN also activated PI3K and Akt, well-known angiogenesis mediators, and the inhibition of either pathway totally abolished the effect of FN on (18)F-FDG uptake. Nitric oxide, a downstream Akt effector that stimulates glucose uptake, was not involved in the metabolic effect of FN. CONCLUSION: The results of this study demonstrated that an EC-FN interaction induces strong enhancement of (18)F-FDG uptake through the upregulation of GLUT1 expression and hexokinase activity. The findings also showed that the response occurs through FAK-mediated activation of PI3K and Akt, indicating a role for this pathway in modulating EC glucose metabolism. PMID- 19289427 TI - The use of 99mTc-recombinant human annexin V imaging for differential diagnosis of aseptic loosening and low-grade infection in hip and knee prostheses. AB - The aim of this study was to assess the pattern of annexin V uptake in hip and knee prostheses suspected of being infected. METHODS: A total of 7 patients undergoing revision surgery for hip or knee prostheses were studied; 5 patients had total hip replacements, and 2 had total knee replacements. Infection was confirmed by pathology, culture results, laboratory evaluation, and clinical follow-up. All patients also underwent a bone scan before surgery. RESULTS: Annexin V scan findings were positive in 5 patients and negative in 2. Annexin V uptake was either focal (n = 4) or linear (n = 1). There were 4 true-positive, 2 true-negative, 1 false-positive, and no false-negative annexin V studies. Annexin V uptake was either more extensive or less extensive than, and usually was incongruent with, (99m)Tc-methylene diphosphonate uptake. CONCLUSION: Our findings suggest that annexin V imaging shows greater uptake with infection than with aseptic loosening and has a high negative predictive value for prosthetic infection. PMID- 19289428 TI - Idiopathic pulmonary fibrosis and diffuse parenchymal lung disease: implications from initial experience with 18F-FDG PET/CT. AB - The purpose of this study was to evaluate integrated (18)F-FDG PET/CT in patients with idiopathic pulmonary fibrosis (IPF) and diffuse parenchymal lung disease (DPLD). METHODS: Thirty-six consecutive patients (31 men and 5 women; mean age +/ SD, 68.7 +/- 9.4 y) with IPF (n = 18) or other forms of DPLD (n = 18) were recruited for PET/CT and high-resolution CT (HRCT), acquired on the same instrument. The maximal pulmonary (18)F-FDG metabolism was measured as a standardized uptake value (SUV(max)). At this site, the predominant lung parenchyma HRCT pattern was defined for each patient: ground-glass or reticulation/honeycombing. Patients underwent a global health assessment and pulmonary function tests. RESULTS: Raised pulmonary (18)F-FDG metabolism in 36 of 36 patients was observed. The parenchymal pattern on HRCT at the site of maximal (18)F-FDG metabolism was predominantly ground-glass (7/36), reticulation/honeycombing (26/36), and mixed (3/36). The mean SUV(max) in patients with ground-glass and mixed patterns was 2.0 +/- 0.4, and in reticulation/honeycombing it was 3.0 +/- 1.0 (Mann-Whitney U test, P = 0.007). The mean SUV(max) in patients with IPF was 2.9 +/- 1.1, and in other DPLD it was 2.7 +/- 0.9 (Mann-Whitney U test, P = 0.862). The mean mediastinal lymph node SUV(max) (2.7 +/- 1.3) correlated with pulmonary SUV(max) (r = 0.63, P < 0.001). Pulmonary (18)F-FDG uptake correlated with the global health score (r = 0.50, P = 0.004), forced vital capacity (r = 0.41, P = 0.014), and transfer factor (r = 0.37, P = 0.042). CONCLUSION: Increased pulmonary (18)F-FDG metabolism in all patients with IPF and other forms of DPLD was observed. Pulmonary (18)F-FDG uptake predicts measurements of health and lung physiology in these patients. (18)F-FDG metabolism was higher when the site of maximal uptake corresponded to areas of reticulation/honeycomb on HRCT than to those with ground-glass patterns. PMID- 19289429 TI - Molecular imaging of matrix metalloproteinase expression in atherosclerotic plaques of mice deficient in apolipoprotein e or low-density-lipoprotein receptor. AB - Matrix metalloproteinases (MMPs) are expressed in atherosclerotic plaques and play an important role in plaque instability. METHODS: Using (99m)Tc-labeled broad-spectrum MMP inhibitor (MPI), we performed noninvasive imaging of MMP expression with micro-SPECT/micro-CT in mice deficient in apolipoprotein E (ApoE( /-), n = 14), mice deficient in low-density-lipoprotein receptor (LDLR(-/-), n = 14), and C57/BL6 mice as controls (n = 7). Seven ApoE(-/-) and 7 LDLR(-/-) received a high-cholesterol diet. After in vivo imaging, aortas were explanted, ex vivo images acquired, and the percent injected dose of MPI per gram (%ID/g) determined, followed by histologic characterization of atherosclerotic lesions. RESULTS: MPI uptake was noninvasively visualized in atherosclerotic lesions by micro-SPECT, with confirmation by micro-CT of anatomic location and aortic calcification. %ID/g in each part of the aorta was highest in ApoE(-/-) that were fed a high-cholesterol diet, followed by LDLR(-/-) that were fed a high cholesterol diet, ApoE(-/-) that were fed normal chow, and LDLR(-/-) that were fed normal chow. The control mice had minimal MPI uptake. A significant correlation was noted between %ID/g and % area positive for macrophages (r = 0.81, P = 0.009), MMP-2 (r = 0.65, P = 0.013), and MMP-9 (r = 0.62, P = 0.008). CONCLUSION: This study demonstrates the usefulness of molecular imaging for noninvasive assessment of the extent of MMP expression in various transgenic mouse models of atherosclerosis receiving a normal or hyperlipidemic diet. It is conceivable that such a strategy may be translationally developed for identification of unstable atherosclerotic plaques. PMID- 19289430 TI - Tissue classification as a potential approach for attenuation correction in whole body PET/MRI: evaluation with PET/CT data. AB - Attenuation correction (AC) of whole-body PET data in combined PET/MRI tomographs is expected to be a technical challenge. In this study, a potential solution based on a segmented attenuation map is proposed and evaluated in clinical PET/CT cases. METHODS: Segmentation of the attenuation map into 4 classes (background, lungs, fat, and soft tissue) was hypothesized to be sufficient for AC purposes. The segmentation was applied to CT-based attenuation maps from (18)F-FDG PET/CT oncologic examinations of 35 patients with 52 (18)F-FDG-avid lesions in the lungs (n = 15), bones (n = 21), and neck (n = 16). The standardized uptake values (SUVs) of the lesions were determined from PET images reconstructed with nonsegmented and segmented attenuation maps, and an experienced observer interpreted both PET images with no knowledge of the attenuation map status. The feasibility of the method was also evaluated with 2 patients who underwent both PET/CT and MRI. RESULTS: The use of a segmented attenuation map resulted in average SUV changes of 8% +/- 3% (mean +/- SD) for bone lesions, 4% +/- 2% for neck lesions, and 2% +/- 3% for lung lesions. The largest SUV change was 13.1%, for a lesion in the pelvic bone. There were no differences in the clinical interpretations made by the experienced observer with both types of attenuation maps. CONCLUSION: A segmented attenuation map with 4 classes derived from CT data had only a small effect on the SUVs of (18)F-FDG-avid lesions and did not change the interpretation for any patient. This approach appears to be practical and valid for MRI-based AC. PMID- 19289431 TI - Imaging of inflamed and vulnerable plaque in coronary arteries with 18F-FDG PET/CT in patients with suppression of myocardial uptake using a low carbohydrate, high-fat preparation. AB - PET/CT imaging with (18)F-FDG has been used to detect inflammation in carotid and aortic plaque; its use in detecting coronary plaque has been limited by avid (18)F-FDG uptake by the myocardium. We investigated whether (18)F-FDG PET/CT could be used to image inflammation in coronary arteries as a potential noninvasive method to detect vulnerable plaque. METHODS: We retrospectively studied 32 patients treated for malignancy who underwent (18)F-FDG PET/CT and concomitant cardiac catheterization. As part of the recently described protocol, all patients were instructed to eat a low-carbohydrate, high-fat meal the night before and drink a vegetable oil drink the morning of the study. We reviewed the patients' baseline characteristics and their (18)F-FDG PET/CT scans for adequacy of myocardial uptake suppression and correlated the presence of angiographically apparent plaque with (18)F-FDG uptake in the major coronary arteries. Two independent observers assessed the angiographic images and (18)F-FDG PET scans. RESULTS: A total of 95% of patients had 2 or more coronary disease risk factors, and 25% had unstable symptoms; 30% of index catheterizations resulted in intervention. In 20 of 32 patients (63%), myocardial suppression was good (12) or adequate (8). Inadequate suppression was due to self-reported dietary nonadherence. Patients with good, adequate, and poor suppression had maximal myocardial standardized uptake values of 2.8 +/- 0.7, 5.0 +/- 1.3, and 17.0 +/- 9.7, respectively. We identified (18)F-FDG uptake in 15 patients in 1 or more coronary segments. A trend to significance in correlation between presence of angiographic disease and signal in the vessel was observed (P = 0.07; 80 vessels examined). A total of 7 patients with significant coronary artery disease had aortic (18)F-FDG uptake. CONCLUSION: In this retrospective study, we demonstrated the potential use of (18)F-FDG PET in imaging of inflammation in coronary arteries. The potential of (18)F-FDG PET is also being investigated in a prospective study. PMID- 19289433 TI - Stress/Rest Myocardial Perfusion Abnormalities by Gated SPECT: Still the Best Predictor of Cardiac Events in Stable Ischemic Heart Disease. AB - The prognostic power of myocardial perfusion imaging in patients with ischemic heart disease (IHD) has been demonstrated since planar imaging. We aimed to investigate whether gated SPECT retains this value in current cardiology if compared with a complete diagnostic work-up and with more recent prognostic indicators. METHODS: We selected from our database a cohort of 676 consecutive inpatients who underwent a complete diagnostic work-up that included gated SPECT and coronary arteriography for known or suspected IHD. Patients with acute myocardial infarction (MI), previous coronary artery bypass surgery, or overt hyperthyroidism and patients who were undergoing dialysis treatment were excluded. During follow-up (median, 37 mo), 24 patients died from cardiac causes and 19 experienced a nonfatal MI. RESULTS: The following were determined to be independent predictors of event-free survival (cardiac death and nonfatal MI) in the different phases of diagnostic work-up using Cox proportional hazards regression analysis: among clinical variables, a previous MI; among laboratory examinations, serum creatinine and low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol levels; among electrocardiographic and echocardiographic variables, left ventricular ejection fraction; and among SPECT variables, summed rest score (SRS) and summed difference score (SDS). In addition, a score of coronary stenoses at angiography was an independent predictor. When the above predictors were tested together, SRS (P < 0.0001), SDS (P = 0.0108), and serum creatinine (P = 0.0186) and LDL and HDL cholesterol levels (P = 0.0222) were the final independent predictors of event-free survival. When gated SPECT was added to the clinical, laboratory, electrocardiographic, and echocardiographic variables, the prognostic stratification significantly improved (P < 0.05); when coronary arteriography was added to gated SPECT, prognostic stratification did not further improve (P > 0.25). If the information provided by gated SPECT was made available after clinical, laboratory, electrocardiographic, echocardiographic, and angiographic variables, the prognostic stratification still improved significantly (P < 0.05). In 492 of these patients with ascertained IHD, SRS and SDS were the final independent predictors of survival. Medical treatment and coronary revascularization did not affect the prognostic information of gated SPECT. CONCLUSION: Myocardial perfusion abnormalities at rest and after stress are still the best predictors of cardiac event-free survival in patients with known or suspected IHD, even when compared with an extensive diagnostic work-up. PMID- 19289434 TI - Quantum dots for in vivo small-animal imaging. AB - Nanotechnology is poised to transform research, prevention, and treatment of cancer through the development of novel diagnostic imaging methods and targeted therapies. In particular, the use of nanoparticles for imaging has gained considerable momentum in recent years. This review focuses on the growing contribution of quantum dots (QDs) for in vivo imaging in small-animal models. Fluorescent QDs, which are small nanocrystals (1-10 nm) made of inorganic semiconductor materials, possess several unique optical properties best suited for in vivo imaging. Because of quantum confinement effects, the emission color of QDs can be precisely tuned by size from the ultraviolet to the near-infrared. QDs are extremely bright and photostable. They are also characterized by a wide absorption band and a narrow emission band, which makes them ideal for multiplexing. Finally, the large surface area of QDs permits the assembly of various contrast agents to design multimodality imaging probes. To date, biocompatible QD conjugates have been used successfully for sentinel lymph node mapping, tumor targeting, tumor angiogenesis imaging, and metastatic cell tracking. Here we consider these novel breakthroughs in light of their potential clinical applications and discuss how QDs might offer a suitable platform to unite disparate imaging modalities and provide information along a continuum of length scales. PMID- 19289435 TI - 99mTc-HYNIC-Gastrin Peptides: Assisted Coordination of 99mTc by Amino Acid Side Chains Results in Improved Performance Both In Vitro and In Vivo. AB - The aim of this study was to determine the effects of assisted coordination by amino acids such as histidine and glutamic acid on the function of (99m)Tc labeled gastrin peptide-hydrazinonicotinamide (HYNIC) conjugates and their ability to target cholecystokinin-R in small-animal models. METHODS: Three peptide-HYNIC conjugates containing the -AYGWMDF-NH2 C-terminal sequence and combinations of histidine, glutamic acid, and glycine were synthesized, radiolabeled with (99m)Tc/(99)Tc using either tricine or ethylenediaminediacetic acid as a coligand, and analyzed by the high-performance liquid chromatography and liquid chromatography-mass spectrometric techniques. Stability, receptor binding, and internalization and in vivo targeting in AR42J-bearing mice were assessed. RESULTS: When radiolabeling was performed using tricine as a coligand, the insertion of a histidine residue near the HYNIC residue resulted in the displacement of one molecule of tricine from the coordination sphere, a reduction in the number of radiolabeled species formed, an improvement in the in vitro stability, an increase in the rate of radiopeptide internalization, and a significant improvement in tumor uptake in vivo. When radiolabeling was performed using ethylenediaminediacetic acid as a coligand, no effect on coligand binding, homogeneity, or in vitro stability was observed but a significant improvement in the internalization in vitro and tumor uptake in vivo was again found. All of the complexes formed showed similar receptor affinity in competitive radioligand binding assays. CONCLUSION: The insertion of histidine into the sequence of peptide-HYNIC conjugates can result in more stable, more homogeneous complexes that show improvements in tumor-targeting performance both in vitro and in vivo. PMID- 19289436 TI - Half-time SPECT myocardial perfusion imaging with attenuation correction. AB - Reducing acquisition time may improve patient throughput, increase camera efficiency, and reduce costs; reducing acquisition time also increases image noise. Newly available software controls the effects of noise by maximum a posteriori reconstruction while maintaining resolution with resolution-recovery methods. This study compares half-time (HT) gated myocardial SPECT images processed with ordered-subset expectation maximization with resolution recovery (OSEM-RR) (with and without CT-based attenuation correction [AC]) with full-time (FT) images obtained with a standard clinical protocol and reconstructed with filtered backprojection (FBP) and OSEM (with and without AC). METHODS: A total of 212 patients (mean age, 57 y; age range, 27-86 y) underwent 1-d rest/stress (99m)Tc-tetrofosmin gated SPECT. FT (12.5 min, both rest and stress) and HT (rest, 7.5 min; stress, 6.0 min) images were acquired with low-dose CT for AC in 112 patients. HT acquisitions were processed with OSEM-RR (with and without AC) using software, and FT acquisitions were processed with FBP and OSEM (with and without AC). In another 100 patients, test-retest repeatability was assessed using 2 sets of FT images (FBP reconstruction) that were acquired one immediately after the other. Radiologists unaware of the acquisition and reconstruction protocols visually assessed all reconstructed images for summed stress, summed rest, and summed difference scores and regional wall motion using a 17-segment model. Automated analysis on gated SPECT was used to determine left ventricular volumes, ejection fraction, and dilation (end-diastolic volume, end-systolic volume, left ventricular ejection fraction, and transient ischemic dilation [TID]). A clinical diagnosis was also determined. RESULTS: All measurements resulted in significant correlations (P < 0.01) between the HT and FT images. The only significant difference in mean values was for OSEM-RR plus AC; this method led to an increase in TID by 4% over FT imaging. The concordance in the clinical diagnosis for HT versus FT was 106 to 112 (kappa = 0.88) for no AC and 102 to 106 (kappa = 0.91) for AC, similar to the repeatability of FT versus FT (98/100, kappa = 0.95). CONCLUSION: HT images processed with the new algorithm provided a clinical diagnosis in concordance with that from FT images in 95% (no AC) to 96% (AC) of cases. This concordance is similar to the test-retest repeatability of FT imaging. PMID- 19289437 TI - One step closer to imaging vulnerable plaque in the coronary arteries. PMID- 19289438 TI - The Role of 18F-FDG PET/CT in the evaluation of Ascites of Undetermined Origin. AB - The first aim of our study was to compare the role of (18)F-FDG PET/CT with that of CT alone in detecting the primary cause of ascites. A secondary aim was to compare the value of (18)F-FDG PET/CT with that of CT alone in detecting abdominal cavity metastasis. Finally, we analyzed the receiver-operating characteristic (ROC) curves of maximal standardized uptake values (SUVmax), serum carcinoembryonic antigen, CA19-9, and CA12-5 for differential diagnostic abilities. METHODS: The (18)F-FDG PET/CT scans of 40 patients with ascites of undetermined origin, including 30 patients with malignant diseases and 10 with benign lesions, were reviewed for the presence of ascites. Among the 40 patients, 34 had received their diagnosis by pathologic examination and 6 by clinical follow-up. We also assessed the (18)F-FDG PET/CT scans of 20 healthy volunteers for comparison. All (18)F-FDG PET/CT images were visually interpreted, and the SUVmax was measured. We compared the mean diameter of true-positive lesions with that of false-negative lesions. The diagnostic abilities of SUVmax, serum carcinoembryonic antigen, CA19-9, and CA12-5 were compared using the ROC curve. RESULTS: The sensitivity, specificity, and accuracy of PET/CT in detecting the primary cause of ascites were 63.3% (19/30), 70.0% (7/10), and 65.0% (26/40), respectively, and those of CT alone were 36.7% (11/30), 80% (8/10), and 47.5% (19/40), respectively (sensitivity, P < 0.05). The sensitivity of PET/CT was higher than that of CT alone for detecting abdominal cavity metastasis (86.4% vs. 27.3%, P < 0.01). The SUVmax in patients with malignant primary and metastatic lesions was significantly higher than that in healthy volunteers and in patients with benign ascites (P < 0.05). The mean maximal diameter of false-negative lesions was significantly smaller than that of true-positive lesions (P < 0.05). In ROC analysis, the areas under the curve of SUVmax, serum carcinoembryonic antigen, CA19-9, and CA12-5 were 0.803 (P < 0.01), 0.773 (P < 0.05), 0.552 (P > 0.05), and 0.220 (P < 0.01), respectively. CONCLUSION: (18)F-FDG PET/CT assisted in detecting the original cause of ascites. The differential diagnostic ability of (18)F-FDG PET/CT was superior to that of CT alone, tumor markers, and cytology. More attention should be paid to peritoneal tuberculosis, which can markedly accumulate (18)F-FDG and mimic peritoneal carcinoma. PMID- 19289439 TI - Novel strategy for a cocktail 18F-fluoride and 18F-FDG PET/CT scan for evaluation of malignancy: results of the pilot-phase study. AB - (18)F-FDG PET/CT is used for detecting cancer and monitoring cancer response to therapy. However, because of the variable rates of glucose metabolism, not all cancers are identified reliably. Sodium (18)F was previously used for bone imaging and can be used as a PET/CT skeletal tracer. The combined administration of (18)F and (18)F-FDG in a single PET/CT study for cancer detection has not been reported to date. METHODS: This is a prospective pilot study (November 2007 November 2008) of 14 patients with proven malignancy (6 sarcoma, 3 prostate cancer, 2 breast cancer, 1 colon cancer, 1 lung cancer, and 1 malignant paraganglioma) who underwent separate (18)F PET/CT and (18)F-FDG PET/CT and combined (18)F/(18)F-FDG PET/CT scans for the evaluation of malignancy (a total of 3 scans each). There were 11 men and 3 women (age range, 19-75 y; average, 50.4 y). RESULTS: Interpretation of the combined (18)F/(18)F-FDG PET/CT scans compared favorably with that of the (18)F-FDG PET/CT (no lesions missed) and the (18)F PET/CT scans (only 1 skull lesion seen on an (18)F PET/CT scan was missed on the corresponding combined scan). Through image processing, the combined (18)F/(18)F-FDG scan yielded results for bone radiotracer uptake comparable to those of the (18)F PET/CT scan performed separately. CONCLUSION: Our pilot-phase prospective trial demonstrates that the combined (18)F/(18)F-FDG administration followed by a single PET/CT scan is feasible for cancer detection. This combined method opens the possibility for improved patient care and reduction in health care costs. PMID- 19289440 TI - MIRD dose estimate report No. 20: radiation absorbed-dose estimates for 111In- and 90Y-ibritumomab tiuxetan. AB - Absorbed-dose calculations provide a scientific basis for evaluating the biologic effects associated with administered radiopharmaceuticals. In cancer therapy, radiation dosimetry supports treatment planning, dose-response analyses, predictions of therapy effectiveness, and completeness of patient medical records. In this study, we evaluated the organ radiation absorbed doses from intravenously administered (111)In- and (90)Y-ibritumomab tiuxetan. METHODS: Ten patients (6 men and 4 women) with non-Hodgkin lymphoma, cared for at 3 different medical centers, were administered the tracer (111)In-ibritumomab tiuxetan and assessed using planar scintillation camera imaging at 5 time points and CT-organ volumetrics to determine patient-specific organ biokinetics and dosimetry. Explicit attenuation correction based on the transmission scan or transmission measurements provided the fraction of (111)In-administered activity in 7 major organs, the whole body, and remainder tissues over time through complete decay. Time-activity curves were constructed, and radiation doses were calculated using MIRD methods and implementing software. RESULTS: Mean radiation absorbed doses for (111)In- and for (90)Y-ibritumomab tiuxetan administered to 10 cancer patients are reported for 24 organs and the whole body. Biologic uptake and retention data are given for 7 major source organs, remainder tissues, and the whole body. Median absorbed dose values calculated by this method were compared with previously published dosimetry for ibritumomab tiuxetan and the product package insert. CONCLUSION: In high-dose radioimmunotherapy, the importance of patient-specific dosimetry becomes obvious when the objective of treatment planning is to achieve disease cures, safely, by limiting radiation dose to any critical normal organ to its maximum tolerable value. Compared with the current package insert, we found differences in median absorbed dose by multiples of 24 in the kidneys, 1.8 in the red marrow, 0.65 in the liver, 0.077 in the intestinal wall, 0.30 in the lungs, 0.46 in the spleen, and 0.34 in the heart wall. PMID- 19289441 TI - SPECT V/Q Scans. PMID- 19289442 TI - A concanavalin A-like lectin domain in the CHS1/LYST protein, shared by members of the BEACH family. AB - CHS1/LYST, the causative protein of the Chediak-Higashi syndrome (CHS), belongs to the BEACH (named after BEige And Chediak-Higashi) family, which includes various large proteins sharing the same C-terminal domain architecture [a PH (Pleckstrin homology)-BEACH domain followed by WD repeats). Members of the BEACH family are generally defined as vesicle-trafficking regulatory proteins, but their functions remain to be determined at the molecular level. Here, using a panel of sensitive methods of sequence analysis, we show that the N-terminal regions of BEACH proteins contain an as yet not described domain, which shares striking similarities with clostridial neurotoxins and defines a novel family within the concanavalin A (ConA)-like lectin superfamily. These results suggest that the BEACH ConA-like lectin domain could be involved in oligosaccharide binding associated with protein traffic and sorting along the secretory pathway, especially in relation with components of the vesicle fusion machinery. PMID- 19289443 TI - A method and program for estimating graphical models for linkage disequilibrium that scale linearly with the number of loci, and their application to gene drop simulation. AB - MOTIVATION: Efficient models for genetic linkage disequilibrium (LD) are needed to enable appropriate statistical analysis of the dense, genome-wide single nucleotide polymorphism assays currently available. RESULTS: Estimation of graphical models for LD within a restricted class of decomposable models is shown to be possible using computer time and storage that scale linearly with the number of loci. Programs for estimation and for simulating from these models on a whole-genome basis are described and provided. AVAILABILITY: Java classes and source code for IntervalLD and GeneDrops are freely available over the internet at http://bioinformatics.med.utah.edu/~alun. PMID- 19289444 TI - GS2: an efficiently computable measure of GO-based similarity of gene sets. AB - MOTIVATION: The growing availability of genome-scale datasets has attracted increasing attention to the development of computational methods for automated inference of functional similarities among genes and their products. One class of such methods measures the functional similarity of genes based on their distance in the Gene Ontology (GO). To measure the functional relatedness of a gene set, these measures consider every pair of genes in the set, and the average of all pairwise distances is calculated. However, as more data becomes available and gene sets used for analysis become larger, such pair-based calculation becomes prohibitive. RESULTS: In this article, we propose GS(2) (GO-based similarity of gene sets), a novel GO-based measure of gene set similarity that is computable in linear time in the size of the gene set. The measure quantifies the similarity of the GO annotations among a set of genes by averaging the contribution of each gene's GO terms and their ancestor terms with respect to the GO vocabulary graph. To study the performance of our method, we compared our measure with an established pair-based measure when run on gene sets with varying degrees of functional similarities. In addition to a significant speed improvement, our method produced comparable similarity scores to the established method. Our method is available as a web-based tool and an open-source Python library. AVAILABILITY: The web-based tools and Python code are available at: http://bioserver.cs.rice.edu/gs2. PMID- 19289445 TI - TopHat: discovering splice junctions with RNA-Seq. AB - MOTIVATION: A new protocol for sequencing the messenger RNA in a cell, known as RNA-Seq, generates millions of short sequence fragments in a single run. These fragments, or 'reads', can be used to measure levels of gene expression and to identify novel splice variants of genes. However, current software for aligning RNA-Seq data to a genome relies on known splice junctions and cannot identify novel ones. TopHat is an efficient read-mapping algorithm designed to align reads from an RNA-Seq experiment to a reference genome without relying on known splice sites. RESULTS: We mapped the RNA-Seq reads from a recent mammalian RNA-Seq experiment and recovered more than 72% of the splice junctions reported by the annotation-based software from that study, along with nearly 20,000 previously unreported junctions. The TopHat pipeline is much faster than previous systems, mapping nearly 2.2 million reads per CPU hour, which is sufficient to process an entire RNA-Seq experiment in less than a day on a standard desktop computer. We describe several challenges unique to ab initio splice site discovery from RNA Seq reads that will require further algorithm development. AVAILABILITY: TopHat is free, open-source software available from http://tophat.cbcb.umd.edu. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19289446 TI - Using multi-data hidden Markov models trained on local neighborhoods of protein structure to predict residue-residue contacts. AB - MOTIVATION: Correct prediction of residue-residue contacts in proteins that lack good templates with known structure would take ab initio protein structure prediction a large step forward. The lack of correct contacts, and in particular long-range contacts, is considered the main reason why these methods often fail. RESULTS: We propose a novel hidden Markov model (HMM)-based method for predicting residue-residue contacts from protein sequences using as training data homologous sequences, predicted secondary structure and a library of local neighborhoods (local descriptors of protein structure). The library consists of recurring structural entities incorporating short-, medium- and long-range interactions and is general enough to reassemble the cores of nearly all proteins in the PDB. The method is tested on an external test set of 606 domains with no significant sequence similarity to the training set as well as 151 domains with SCOP folds not present in the training set. Considering the top 0.2 x L predictions (L = sequence length), our HMMs obtained an accuracy of 22.8% for long-range interactions in new fold targets, and an average accuracy of 28.6% for long-, medium- and short-range contacts. This is a significant performance increase over currently available methods when comparing against results published in the literature. AVAILABILITY: http://predictioncenter.org/Services/FragHMMent/. PMID- 19289447 TI - Transcriptionally active gene fragments derived from potentially fast-evolving donor genes in the rice genome. AB - The unprecedented complexity of the transcriptomic data obtained in recent years creates opportunities for new genomic studies aimed at interpolating regulatory code of gene expression and tracing genome evolution. We report here the identification and characterization of a set of 851 intergenic loci that represent transcribed gene fragments (TGFs) ectopically duplicated from 1030 non transposable element (non-TE) donor genes in the rice genome. We analyzed the genomic context of the TGFs and donor genes. We show that the TGFs have adopted transcriptional orientation and pattern independent of the donor genes. We further show that TGFs have undergone relaxed purifying selection, consistent with their being pseudogenized. We found that the donor genes, which are biased toward certain molecular functions, exhibit an accelerated evolution rate comparing to the genome average. Our results demonstrated a large number of actively TGFs in the rice genome and shed light on the origin, mode of action and function of the TGFs. PMID- 19289448 TI - Gaussian process regression bootstrapping: exploring the effects of uncertainty in time course data. AB - MOTIVATION: Although widely accepted that high-throughput biological data are typically highly noisy, the effects that this uncertainty has upon the conclusions we draw from these data are often overlooked. However, in order to assign any degree of confidence to our conclusions, we must quantify these effects. Bootstrap resampling is one method by which this may be achieved. Here, we present a parametric bootstrapping approach for time-course data, in which Gaussian process regression (GPR) is used to fit a probabilistic model from which replicates may then be drawn. This approach implicitly allows the time dependence of the data to be taken into account, and is applicable to a wide range of problems. RESULTS: We apply GPR bootstrapping to two datasets from the literature. In the first example, we show how the approach may be used to investigate the effects of data uncertainty upon the estimation of parameters in an ordinary differential equations (ODE) model of a cell signalling pathway. Although we find that the parameter estimates inferred from the original dataset are relatively robust to data uncertainty, we also identify a distinct second set of estimates. In the second example, we use our method to show that the topology of networks constructed from time-course gene expression data appears to be sensitive to data uncertainty, although there may be individual edges in the network that are robust in light of present data. AVAILABILITY: Matlab code for performing GPR bootstrapping is available from our web site: http://www3.imperial.ac.uk/theoreticalsystemsbiology/data-software/. PMID- 19289449 TI - Early weight gain predicts retinopathy in preterm infants: new, simple, efficient approach to screening. AB - BACKGROUND: The risk for sight-threatening retinopathy of prematurity is predicted by using gestational age and/or weight at birth. All infants below a threshold undergo serial ophthalmologic examinations for identification of those who would benefit from treatment (approximately 10%). We hypothesized that factoring in postnatal weight gain could identify children at risk for sight threatening retinopathy of prematurity more specifically and earlier. METHODS: Weekly weights from birth to postmenstrual week 36 were retrospectively entered into a surveillance system that gave an alarm when the rate of weight gain decreased to a certain level. For all children (N = 354) screened and/or treated for retinopathy of prematurity at Sahlgrenska University Hospital in 2004-2007, weekly weights were recorded. One child was excluded because of known nonphysiologic weight gain (hydrocephalus). RESULTS: For 127 (36%) of 353 children, no alarm was given; for 40%, alarm at low risk was given after postmenstrual week 32. None of those children developed retinopathy of prematurity requiring treatment. Of the remaining 24% of children who received alarm at high or low risk before 32 postmenstrual weeks, 41% developed proliferative retinopathy of prematurity and 29% were treated because of sight threatening disease. The median time from alarm to treatment was 9 weeks. CONCLUSIONS: The weight, insulin-like growth factor, neonatal retinopathy of prematurity algorithm detected early 100% of infants who developed retinopathy of prematurity requiring treatment and correctly predicted the majority who did not require treatment. With this simple postnatal evaluation, costly stressful eye examinations can be markedly reduced (approximately 75% of infants). In addition, early identification of children at risk may lead to the initiation of interventions and possibly prevent sight-threatening retinopathy of prematurity. PMID- 19289450 TI - Intravenous ceftriaxone and calcium in the neonate: assessing the risk for cardiopulmonary adverse events. AB - OBJECTIVES: Unsolicited reports regarding potentially serious adverse drug reactions in neonates and young infants were reported to the Food and Drug Administration, leading to changes in the package label for ceftriaxone. This report describes and summarizes the reported cases that led to safety concerns regarding the concurrent administration of intravenous ceftriaxone and calcium in this age group. METHODS: Nine reported cases were assessed. The Food and Drug Administration Adverse Event Reporting System database was searched for potential drug interactions in patients who were receiving concomitant ceftriaxone and calcium therapy. RESULTS: Eight of the reported 9 cases (7 were < or =2 months of age) represented possible or probable adverse drug events. There were 7 deaths. None of the cases were reported from the United States. The dosage of ceftriaxone that was administered to 4 of 6 infants for whom this information was available was between 150 and 200 mg/kg per day. The rate of occurrence of these serious adverse drug reactions cannot be accurately determined from available data. CONCLUSIONS: The concurrent use of intravenous ceftriaxone and calcium-containing solutions in the newborn and young infant may result in a life-threatening adverse drug reaction. Contributing factors for infants in this report may include the use of ceftriaxone at dosages higher than those approved by the Food and Drug Administration, intravenous "push" administration, and administration of the total daily dosage as a single infusion. PMID- 19289451 TI - Major depression and antidepressant treatment: impact on pregnancy and neonatal outcomes. AB - OBJECTIVE: Selective serotonin reuptake inhibitor (SSRI) use during pregnancy incurs a low absolute risk for major malformations; however, other adverse outcomes have been reported. Major depression also affects reproductive outcomes. This study examined whether 1) minor physical anomalies, 2) maternal weight gain and infant birth weight, 3) preterm birth, and 4) neonatal adaptation are affected by SSRI or depression exposure. METHOD: This prospective observational investigation included maternal assessments at 20, 30, and 36 weeks of gestation. Neonatal outcomes were obtained by blinded review of delivery records and infant examinations. Pregnant women (N=238) were categorized into three mutually exclusive exposure groups: 1) no SSRI, no depression (N=131); 2) SSRI exposure (N=71), either continuous (N=48) or partial (N=23); and 3) major depressive disorder (N=36), either continuous (N=14) or partial (N=22). The mean depressive symptom level of the group with continuous depression and no SSRI exposure was significantly greater than for all other groups, demonstrating the expected treatment effect of SSRIs. Main outcomes were minor physical anomalies, maternal weight gain, infant birth weight, pregnancy duration, and neonatal characteristics. RESULTS: Infants exposed to either SSRIs or depression continuously across gestation were more likely to be born preterm than infants with partial or no exposure. Neither SSRI nor depression exposure increased risk for minor physical anomalies or reduced maternal weight gain. Mean infant birth weights were equivalent. Other neonatal outcomes were similar, except 5-minute Apgar scores. CONCLUSIONS: For depressed pregnant women, both continuous SSRI exposure and continuous untreated depression were associated with preterm birth rates exceeding 20%. PMID- 19289452 TI - Altered markers of tonic inhibition in the dorsolateral prefrontal cortex of subjects with schizophrenia. AB - OBJECTIVE: Cognitive impairments in schizophrenia are associated with lower expression of markers of gamma-aminobutyric acid (GABA) synthesis in the prefrontal cortex. The effects of GABA are mediated by GABA(A) receptors that mediate either phasic or tonic inhibition. The authors assessed the expression of GABA(A) receptor alpha4 and delta subunits, which coassemble to form receptors mediating tonic inhibition, in schizophrenia. METHOD: The authors used in situ hybridization to quantify expression patterns of GABA(A) receptor alpha4 and delta subunits in prefrontal cortex from 23 matched pairs of schizophrenia and comparison subjects. RESULTS: Levels of delta mRNA were significantly lower in schizophrenia subjects regardless of medication use, whereas alpha4 mRNA levels were lower only in subjects with schizophrenia receiving certain medications at the time of death. To understand the nature of this unexpected dissociation between alpha4 and delta subunit expression in schizophrenia, the authors used similar methods to quantify alpha4 and delta mRNA levels in multiple animal models. During postnatal development of monkey prefrontal cortex, levels of alpha4 mRNA decreased, whereas delta mRNA levels increased. In addition, delta mRNA levels, but not alpha4 mRNA levels, were lower in the medial frontal cortex of mice with a genetic deletion of the GABA(A) receptor alpha1 subunit, and neither delta nor alpha4 mRNA levels were altered in rodent models of altered excitatory neurotransmission. CONCLUSIONS: Since GABA(A) receptor alpha1 subunits also have lower mRNA levels in schizophrenia, show increased expression with age in monkey prefrontal cortex, and can coassemble with delta subunits to form functional GABA(A) receptors, lower delta mRNA levels in schizophrenia might reflect a reduced number of alpha(1)beta(x)delta GABA(A) receptors that could contribute to deficient tonic inhibition and prefrontal cortical dysfunction in schizophrenia. PMID- 19289453 TI - Can clinicians recognize DSM-IV personality disorders from five-factor model descriptions of patient cases? AB - OBJECTIVE: This article examined, using theories from cognitive science, the clinical utility of the Five-Factor Model (FFM) of Personality, an assessment and classification system under consideration for integration into the forthcoming fifth edition of the Diagnostic and Statistical Manual (DSM) of Mental Disorders. Specifically, the authors sought to test whether FFM descriptors are specific enough to allow practicing clinicians to capture core features of personality disorders. METHOD: In two studies, a large nationwide sample of clinical psychologists, psychiatrists, and clinical social workers (N=187 and N=191) were presented case profiles based on symptom formats from either the DSM-IV and/or FFM. Ratings for six aspects of clinical utility for DSM-IV and FFM profiles were obtained and participants provided DSM-IV diagnoses. Prototypic cases (only one personality disorder) and comorbid cases were tested in separate studies. RESULTS: Participants rated the DSM-IV as more clinically useful than the FFM on five out of six clinical utility questions. Despite demonstrating considerable background knowledge of DSM-IV diagnoses, participants had difficulty identifying correct diagnoses from FFM profiles. CONCLUSION: The FFM descriptors may be more ambiguous than the criteria of the DSM-IV and the FFM may therefore be less able to convey important clinical details than the DSM-IV. The findings flag challenges to clinical utility for dimensional-trait systems such as the FFM. PMID- 19289454 TI - Maintenance treatment for patients with bipolar I disorder: results from a north american study of quetiapine in combination with lithium or divalproex (trial 127). AB - OBJECTIVE: The authors evaluated the efficacy and safety of quetiapine plus lithium or divalproex in the prevention of recurrent mood events in patients with stabilized bipolar I disorder. METHOD: A total of 1,953 patients received open label quetiapine (400-800 mg/day in flexible, divided doses) with either lithium or divalproex (target serum concentrations 0.5-1.2 meq/liter and 50-125 microg/ml, respectively) for up to 36 weeks. After at least 12 weeks of clinical stability, 628 patients were randomly assigned to double-blind treatment with quetiapine or placebo, in combination with lithium or divalproex, for up to 104 weeks. The primary efficacy measure was time to recurrence of any mood event (mania, depression, or a mixed episode). RESULTS: Fewer patients in the quetiapine group experienced a mood event compared with the placebo group (20.3% versus 52.1%). The hazard ratio for time to recurrence of a mood event was 0.32. Hazard ratios were similar for mania and depression events (0.30 and 0.33, respectively). Sedation, weight increase, and hypothyroidism occurred more frequently in the quetiapine group, as did discontinuations due to adverse events. The incidence and incidence density of a single emergent blood glucose value > or =126 mg/dl were higher in the quetiapine group (12.6% versus 5.4%; 18.44 versus 9.56 patients per 100 patient-years). Adverse events were generally consistent with the known tolerability profile of quetiapine. CONCLUSIONS: In patients stabilized on quetiapine plus lithium or divalproex, continued treatment was associated with a significant risk reduction in the time to recurrence of any mood event compared with placebo and lithium or divalproex. PMID- 19289455 TI - Treatment-resistant depression and mortality after acute coronary syndrome. AB - Depression is a risk factor for morbidity and mortality in patients with coronary heart disease, especially following acute coronary syndrome. Evidence from recent clinical trials suggests that treatment-resistant depression may be associated with a particularly high risk of mortality or cardiac morbidity in patients following acute coronary syndrome. This article reviews this evidence and considers possible explanations for this relationship. Directions for future research are also considered, with particular emphasis on efforts to elucidate the underlying mechanisms and to develop more efficacious treatments for depression in patients with coronary heart disease. PMID- 19289456 TI - Diabetic retinal neurodegeneration is associated with mitochondrial oxidative stress and is improved by an angiotensin receptor blocker in a model combining hypertension and diabetes. AB - OBJECTIVE: Diabetic retinopathy displays the features of a neurodegenerative disease. Oxidative stress is involved in the pathogenesis of diabetic retinopathy. This investigation sought to determine whether hypertension exacerbates the oxidative stress, neurodegeneration, and mitochondrial dysfunction that exists in diabetic retinopathy and whether these changes could be minimized by the angiotensin II type 1 (AT(1)) receptor blocker (ARB) losartan. RESEARCH DESIGN AND METHODS: Diabetes was induced in spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats. The diabetic SHRs were assigned to receive or not receive losartan. RESULTS: The level of apoptosis in the retina was higher in diabetic WKY rats than in the control group, and higher levels were found in diabetic SHRs. The apoptotic cells expressed neural and glial markers. The retinal glial reaction was more evident in diabetic WKY rats and was markedly accentuated in diabetic SHRs. Superoxide production in retinal tissue increased in diabetic WKY rats, and a greater increase occurred in diabetic SHRs. Glutathione levels decreased only in diabetic SHRs. As a consequence, the levels of nitrotyrosine and 8-hydroxy 2' deoxyguanosine, markers of oxidative stress, were elevated in diabetic groups, mainly in diabetic SHRs. Mitochondrial integrity was dramatically affected in the diabetic groups. The ARB treatment reestablished all of the above-mentioned parameters. CONCLUSIONS: These findings suggest that concomitance of hypertension and diabetes exacerbates oxidative stress, neurodegeneration, and mitochondrial dysfunction in the retinal cells. These data provide the first evidence of AT(1)blockage as a neuroprotective treatment of diabetic retinopathy by reestablishing oxidative redox and the mitochondrial function. PMID- 19289457 TI - Inhibition of Th17 cells regulates autoimmune diabetes in NOD mice. AB - OBJECTIVE: The T helper 17 (Th17) population, a subset of CD4-positive T-cells that secrete interleukin (IL)-17, has been implicated in autoimmune diseases, including multiple sclerosis and lupus. Therapeutic agents that target the Th17 effector molecule IL-17 or directly inhibit the Th17 population (IL-25) have shown promise in animal models of autoimmunity. The role of Th17 cells in type 1 diabetes has been less clear. The effect of neutralizing anti-IL-17 and recombinant IL-25 on the development of diabetes in NOD mice, a model of spontaneous autoimmune diabetes, was investigated in this study. RESEARCH DESIGN AND METHODS AND RESULTS: Although treatment with either anti-IL-17 or IL-25 had no effect on diabetes development in young (<5 weeks) NOD mice, either intervention prevented diabetes when treatment was started at 10 weeks of age (P < 0.001). Insulitis scoring and immunofluorescence staining revealed that both anti-IL-17 and IL-25 significantly reduced peri-islet T-cell infiltrates. Both treatments also decreased GAD65 autoantibody levels. Analysis of pancreatic lymph nodes revealed that both treatments increased the frequency of regulatory T cells. Further investigation demonstrated that IL-25 therapy was superior to anti IL-17 during mature diabetes because it promoted a period of remission from new onset diabetes in 90% of treated animals. Similarly, IL-25 delayed recurrent autoimmunity after syngeneic islet transplantation, whereas anti-IL-17 was of no benefit. GAD65-specific ELISpot and CD4-positive adoptive transfer studies showed that IL-25 treatment resulted in a T-cell-mediated dominant protective effect against autoimmunity. CONCLUSIONS: These studies suggest that Th17 cells are involved in the pathogenesis of autoimmune diabetes. Further development of Th17 targeted therapeutic agents may be of benefit in this disease. PMID- 19289458 TI - FoxO1 haploinsufficiency protects against high-fat diet-induced insulin resistance with enhanced peroxisome proliferator-activated receptor gamma activation in adipose tissue. AB - OBJECTIVE: Forkhead box O (FoxO) transcription factors represent evolutionarily conserved targets of insulin signaling, regulating metabolism and cellular differentiation in response to changes in nutrient availability. Although the FoxO1 isoform is known to play a key role in adipogenesis, its physiological role in differentiated adipose tissue remains unclear. RESEARCH DESIGN AND METHODS: In this study, we analyzed the phenotype of FoxO1 haploinsufficient mice to investigate the role of FoxO1 in high-fat diet-induced obesity and adipose tissue metabolism. RESULTS: We showed that reduced FoxO1 expression protects mice against obesity-related insulin resistance with marked improvement not only in hepatic insulin sensitivity but also in skeletal muscle insulin action. FoxO1 haploinsufficiency also resulted in increased peroxisome proliferator-activated receptor (PPAR)gamma gene expression in adipose tissue, with enhanced expression of PPARgamma target genes known to influence metabolism. Moreover, treatment of mice with the PPARgamma agonist rosiglitazone caused a greater improvement in in vivo insulin sensitivity in FoxO1 haploinsufficient animals, including reductions in circulating proinflammatory cytokines. CONCLUSIONS: These findings indicate that FoxO1 proteins negatively regulate insulin action and that their effect may be explained, at least in part, by inhibition of PPARgamma function. PMID- 19289459 TI - I do not smoke -- I exercise: a pilot study of a new educational resource for secondary education students. AB - AIMS: The aim of this study was to examine the applicability of a smoking prevention educational resource for secondary education students named ;I do not smoke, I exercise'. The main focus of this program was the promotion of exercise as an alternative behaviour to smoking. METHODS: The applicability of the intervention was assessed through questionnaires regarding: (a) attitudes towards smoking, (b) interest in information about smoking, and (c) knowledge about health consequences of smoking. Moreover, interviews were taken from the educators and the school principals. The questionnaires were completed from 210 students (101 boys and 109 girls) before the beginning, at the end of the program, and 12 months after the completion of the programme. One-way repeated measure ANOVA was performed to examine the differences between the pre-, post-, and the follow-up measures. RESULTS: Results showed that the program succeeded in changing the students' attitudes, but the follow-up measures, 12 months later, showed that attitudes towards smoking and interest in information were relapsed to the pre-intervention levels. The only effect that was sustained was that for knowledge. The interviews also revealed positive comments about the program implementation and pointed some shortcomings. CONCLUSIONS: The program needs some modifications in order to overcome its weaknesses, have more lasting effects and to improve its applicability. PMID- 19289460 TI - Tetrameric Orai1 is a teardrop-shaped molecule with a long, tapered cytoplasmic domain. AB - The Ca(2+) release-activated Ca(2+) channel is a principal regulator of intracellular Ca(2+) rise, which conducts various biological functions, including immune responses. This channel, involved in store-operated Ca(2+) influx, is believed to be composed of at least two major components. Orai1 has a putative channel pore and locates in the plasma membrane, and STIM1 is a sensor for luminal Ca(2+) store depletion in the endoplasmic reticulum membrane. Here we have purified the FLAG-fused Orai1 protein, determined its tetrameric stoichiometry, and reconstructed its three-dimensional structure at 21-A resolution from 3681 automatically selected particle images, taken with an electron microscope. This first structural depiction of a member of the Orai family shows an elongated teardrop-shape 150A in height and 95A in width. Antibody decoration and volume estimation from the amino acid sequence indicate that the widest transmembrane domain is located between the round extracellular domain and the tapered cytoplasmic domain. The cytoplasmic length of 100A is sufficient for direct association with STIM1. Orifices close to the extracellular and intracellular membrane surfaces of Orai1 seem to connect outside the molecule to large internal cavities. PMID- 19289461 TI - The acute lymphoblastic leukemia-associated JAK2 L611S mutant induces tumorigenesis in nude mice. AB - JAK2 plays important roles in the regulation of a variety of cellular processes including cell migration, proliferation, and protection from apoptosis. Recently the L611S point mutation in JAK2 has been identified in a child with acute lymphoblastic leukemia. Here we analyzed the mechanism by which JAK2 exhibits its oncogenicity. In BaF3 murine hematopoietic cells, L611S mutant increased the expression of antiapoptotic proteins including X chromosome-linked inhibitor of apoptosis protein, inhibitor of apoptosis protein, and Bcl-XL. We also showed that JAK2 L611S mutant protects BaF3 cells from cytokine withdrawal-induced apoptotic cell death and leads to cytokine-independent cell growth. Furthermore BaF3 cells expressing JAK2 L611S mutant gained the ability to induce tumorigenesis in nude mice. The L611S mutant also exhibited malignancy, including prompt invasion and spreading into various organs, leading to rapid lethality of the mice. Finally we showed that a specific JAK2 inhibitor, AG490, potently inhibited cytokine-independent cell growth induced by JAK2 L611S mutant via the induction of apoptotic cell death. In addition, treatment with AG490 significantly inhibited the JAK2 L611S mutant-induced tumorigenesis in nude mice. Thus, our results both in vitro and in vivo strongly suggest that L611S mutant of JAK2 harbors potent oncogenic activity, and this probably requires the antiapoptotic signaling pathway. PMID- 19289462 TI - Leucine/valine residues direct oxygenation of linoleic acid by (10R)- and (8R) dioxygenases: expression and site-directed mutagenesis oF (10R)-dioxygenase with epoxyalcohol synthase activity. AB - Linoleate (10R)-dioxygenase (10R-DOX) of Aspergillus fumigatus was cloned and expressed in insect cells. Recombinant 10R-DOX oxidized 18:2n-6 to (10R) hydroperoxy-8(E),12(Z)-octadecadienoic acid (10R-HPODE; approximately 90%), (8R) hydroperoxylinoleic acid (8R-HPODE; approximately 10%), and small amounts of 12S(13R)-epoxy-(10R)-hydroxy-(8E)-octadecenoic acid. We investigated the oxygenation of 18:2n-6 at C-10 and C-8 by site-directed mutagenesis of 10R-DOX and 7,8-linoleate diol synthase (7,8-LDS), which forms approximately 98% 8R-HPODE and approximately 2% 10R-HPODE. The 10R-DOX and 7,8-LDS sequences differ in homologous positions of the presumed dioxygenation sites (Leu-384/Val-330 and Val 388/Leu-334, respectively) and at the distal site of the heme (Leu-306/Val-256). Leu-384/Val-330 influenced oxygenation, as L384V and L384A of 10R-DOX elevated the biosynthesis of 8-HPODE to 22 and 54%, respectively, as measured by liquid chromatography-tandem mass spectrometry analysis. The stereospecificity was also decreased, as L384A formed the R and S isomers of 10-HPODE and 8-HPODE in a 3:2 ratio. Residues in this position also influenced oxygenation by 7,8-LDS, as its V330L mutant augmented the formation of 10R-HPODE 3-fold. Replacement of Val-388 in 10R-DOX with leucine and phenylalanine increased the formation of 8R-HPODE to 16 and 36%, respectively, whereas L334V of 7,8-LDS was inactive. Mutation of Leu 306 with valine or alanine had little influence on the epoxyalcohol synthase activity. Our results suggest that Leu-384 and Val-388 of 10R-DOX control oxygenation of 18:2n-6 at C-10 and C-8, respectively. The two homologous positions of prostaglandin H synthase-1, Val-349 and Ser-353, are also critical for the position and stereospecificity of the cyclooxygenase reaction. PMID- 19289463 TI - 14-3-3zeta contributes to tyrosine hydroxylase activity in MN9D cells: localization of dopamine regulatory proteins to mitochondria. AB - The 14-3-3 proteins stimulate the activation of tyrosine hydroxylase (TH), the rate-limiting catecholamine biosynthetic enzyme. To explore if particular endogenous 14-3-3 isoforms specifically affected TH activity and dopamine synthesis, we utilized rodent nigrostriatal tissues and midbrain-derived MN9D dopaminergic cells. Extracts from ventral midbrain and MN9D cells contained similar pools of 14-3-3 mRNAs, with 14-3-3zeta being relatively abundant in both. Protein levels of 14-3-3zeta were also abundant. [(32)P]Orthophosphate labeling of MN9D cells, followed by co-immunoprecipitation with pan-TH and pan-14-3-3 antibodies brought down similar amounts of phosphorylated TH in each, confirming that 14-3-3-bound phosphorylated TH in our cells. Co-immunoprecipitation of striatal tissues with a pan-TH antibody precipitated 14-3-3zeta but not another potential TH regulatory isoform, 14-3-3eta. In whole cell extracts from MN9D cells after 14-3-3 small interfering RNA treatments, we found that 14-3-3zeta knockdown significantly reduced TH activity and dopamine synthesis whereas knockdown of 14-3-3eta had no effect. 14-3-3zeta was found co-localized on mitochondria with TH, and its knockdown by small interfering RNA reduced TH phosphorylation and TH activity in the mitochondrial pool. Together the data support a role for 14-3-3zeta as an endogenous activator of TH in midbrain dopaminergic neurons and furthermore, identify mitochondria as a potential novel site for dopamine synthesis, with implications for Parkinson disease. PMID- 19289464 TI - Dynamic Organization of Aminoacyl-tRNA Synthetase Complexes in the Cytoplasm of Human Cells. AB - The localization in space and in time of proteins within the cytoplasm of eukaryotic cells is a central question of the cellular compartmentalization of metabolic pathways. The assembly of proteins within stable or transient complexes plays an essential role in this process. Here, we examined the subcellular localization of the multi-aminoacyl-tRNA synthetase complex in human cells. The sequestration of its components within the cytoplasm rests on the presence of the eukaryotic-specific polypeptide extensions that characterize the human enzymes, as compared with their prokaryotic counterparts. The cellular mobility of several synthetases, assessed by measuring fluorescence recovery after photobleaching, suggested that they are not freely diffusible within the cytoplasm. Several of these enzymes, isolated by tandem affinity purification, were copurified with ribosomal proteins and actin. The capacity of aminoacyl-tRNA synthetases to interact with polyribosomes and with the actin cytoskeleton impacts their subcellular localization and mobility. Our observations have conceptual implications for understanding how translation machinery is organized in vivo. PMID- 19289465 TI - Caspase-mediated cleavage of beta-catenin precedes drug-induced apoptosis in resistant cancer cells. AB - A delicate balance between cell death and survival pathways maintains normal physiology, which is altered in many cancers, shifting the balance toward increased survival. Several studies have established a close connection between the Wnt/beta-catenin pathway and tumorigenesis, aberrant activation of which might contribute toward increased cancer cell growth and survival. Extensive research is underway to identify therapeutic agents that can induce apoptosis specifically in cancer cells with minimal collateral damage to normal cells. Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can induce apoptosis specifically in tumor cells, many cancer cells develop resistance, which can be overcome by combinatorial treatment with other agents: for example, peroxisome proliferator-activated receptor gamma (PPARgamma) ligands. To identify the molecular target mediating combinatorial drug-induced apoptosis, we focused on beta-catenin, a protein implicated in oncogenesis. Our results show that co-treatment of TRAIL-resistant cancer cells with TRAIL and the PPARgamma ligand troglitazone leads to a reduction of beta-catenin expression, coinciding with maximal apoptosis. Modulation of beta-catenin levels via ectopic overexpression or small interference RNA-mediated gene silencing modulates drug induced apoptosis, indicating involvement of beta-catenin in regulating this pathway. More in-depth studies indicated a post-translational mechanism, independent of glycogen synthase kinase-3beta activity regulating beta-catenin expression following combinatorial drug treatment. Furthermore, TRAIL- and troglitazone-induced apoptosis was preceded by a cleavage of beta-catenin, which was complete in a fully apoptotic population, and was mediated by caspases-3 and 8. These results demonstrate beta-catenin as a promising new target of drug induced apoptosis, which can be targeted to sensitize apoptosis-resistant cancer cells. PMID- 19289466 TI - Structure of the 12-subunit RNA polymerase II refined with the aid of anomalous diffraction data. AB - RNA polymerase II (Pol II) is the central enzyme of eukaryotic gene expression machinery. Complete definition of the three-dimensional structure of Pol II is essential for understanding the mechanisms that regulate transcription via protein-protein interactions within the Pol II apparatus. To date a series of Pol II-related crystal structures have been reported. However, certain peptide regions, including several that are implicated to interact with regulatory factors, remain obscure. Here we describe conformations for two such regions that are close to the Pol II surface and assume seemingly flexible loop structures. One is located in the TFIIF-interacting Protrusion domain, whereas the other is in the TFIIE-interacting Clamp domain. This structural definition was aided by the application of an advanced crystallographic refinement approach that utilizes the single anomalous diffraction (SAD) from zinc ions bound intrinsically in Pol II. The SAD-based strategy allowed the 12-subunit Pol II model to be fully refined up to 3.8 A with excellent stereochemical properties, demonstrating the effectiveness of the SAD approach for the refinement of large structures at low to-moderate resolutions. Our results also define additional components of the free Pol II, including the functionally critical Fork Loop-1 and Fork Loop-2 elements. As such, this refined Pol II model provides the most complete structural reference for future analyses of complex structures formed between Pol II and its regulatory factors. PMID- 19289467 TI - Indirect regulation of presenilins in CREB-mediated transcription. AB - Presenilins are essential for synaptic function, memory formation, and neuronal survival. Previously, we reported that expression of cAMP response element binding protein (CREB) target genes is reduced in the cerebral cortex of presenilin (PS) conditional double knock-out (cDKO) mice. To determine whether the reduced expression of the CREB target genes in these mutant mice is due to loss of presenilin directly or secondary to the impaired neuronal activity, we established a sensitive luciferase reporter system to assess direct transcriptional regulation in cultured cells. We first used immortalized PS deficient mouse embryonic fibroblasts (MEFs), and found that both CREB-mediated transcription and Notch-mediated HES1 transcription are decreased. However, the ubiquitin-C promoter-mediated transcription is also reduced, and among these three reporters, transfection of exogenous PS1 can rescue only the Notch-mediated HES1 transcription. Further Northern analysis revealed transcriptional alterations of Creb, ubiquitin-C, and other housekeeping genes in PS-deficient MEFs, indicating transcriptional dysregulation in these cells. We then used the Cre/loxP system to develop a postnatal PS-deficient cortical neuronal culture. Surprisingly, in these PS-null neurons, CREB-mediated transcription is not significantly decreased, and levels of total and phosphorylated CREB proteins are unchanged as well. Notch-mediated HES1 transcription is markedly reduced, and this reduction can be rescued by exogenous PS1. Together, our findings suggest that CREB-mediated transcription is regulated indirectly by PS in the adult cerebral cortex, and that attenuation of CREB target gene expression in PS cDKO mice is likely due to reduced neuronal activity in these mutant brains. PMID- 19289468 TI - MADD, a splice variant of IG20, is indispensable for MAPK activation and protection against apoptosis upon tumor necrosis factor-alpha treatment. AB - We investigated the physiological role of endogenous MAPK-activating death domain containing protein (MADD), a splice variant of the IG20 gene, that can interact with TNFR1 in tumor necrosis factor-alpha (TNFalpha)-induced activation of NF kappaB, MAPK, ERK1/2, JNK, and p38. Using exon-specific short hairpin RNAs expressing lentiviruses, we knocked down the expression of all IG20 splice variants or MADD, which is overexpressed in cancer cells. Abrogation of MADD expression rendered cells highly susceptible to TNFalpha-induced apoptosis in the absence of cycloheximide. It also resulted in a dramatic loss in TNFalpha-induced activation of MAPK without any apparent effect on NF-kappaB activation. This observation was substantiated by an accompanying loss in the activation of p90RSK, a key downstream target of MAPK, whereas the NF-kappaB-regulated interleukin 6 levels remained unaffected. Endogenous MADD knockdown, however, did not affect epidermal growth factor-induced MAPK activation thereby demonstrating the specific requirement of MADD for TNF receptor-mediated MAPK activation. Re expression of short hairpin RNA-resistant MADD in the absence of endogenous IG20 expression rescued the cells from TNFalpha-induced apoptosis. The requirement for MADD was highly specific for TNFalpha-induced activation of MAPK but not the related JNK and p38 kinases. Loss of MADD expression resulted in reduced Grb2 and Sos1/2 recruitment to the TNFR1 complex and decreased Ras and MEKK1/2 activation. These results demonstrate the essential role of MADD in protecting cancer cells from TNFalpha-induced apoptosis by specifically activating MAPKs through Grb2 and Sos1/2 recruitment, and its potential as a novel cancer therapeutic target. PMID- 19289469 TI - Generating an unfoldase from thioredoxin-like domains. AB - Protein-disulfide isomerase (PDI), an endoplasmic reticulum (ER)-resident protein, is primarily known as a catalyst of oxidative protein folding but also has a protein unfolding activity. We showed previously that PDI unfolds the cholera toxin A1 (CTA1) polypeptide to facilitate the ER-to-cytosol retrotranslocation of the toxin during intoxication. We now provide insight into the mechanism of this unfoldase activity. PDI includes two redox-active (a and a') and two redox-inactive (b and b') thioredoxin-like domains, a linker (x), and a C-terminal domain (c) arranged as abb'xa'c. Using recombinant PDI fragments, we show that binding of CTA1 by the continuous PDIbb'xa' fragment is necessary and sufficient to trigger unfolding. The specific linear arrangement of bb'xa' and the type a domain (a' versus a) C-terminal to bb'x are additional determinants of activity. These data suggest a general mechanism for the unfoldase activity of PDI: the concurrent and specific binding of bb'xa' to particular regions along the CTA1 molecule triggers its unfolding. Furthermore, we show the bb' domains of PDI are indispensable to the unfolding reaction, whereas the function of its a' domain can be substituted partially by the a' domain from ERp57 (abb'xa'c) or ERp72 (ca degrees abb'xa'), PDI-like proteins that do not unfold CTA1 normally. However, the bb' domains of PDI were insufficient to convert full-length ERp57 into an unfoldase because the a domain of ERp57 inhibited toxin binding. Thus, we propose that generating an unfoldase from thioredoxin-like domains requires the bb'(x) domains of PDI followed by an a' domain but not preceded by an inhibitory a domain. PMID- 19289470 TI - Promotion of neurite extension by protrudin requires its interaction with vesicle associated membrane protein-associated protein. AB - Protrudin is a protein that contains a Rab11-binding domain and a FYVE (lipid binding) domain and that functions to promote neurite formation through interaction with the GDP-bound form of Rab11. Protrudin also contains a short sequence motif designated FFAT (two phenylalanines in an acidic tract), which in other proteins has been shown to mediate binding to vesicle-associated membrane protein-associated protein (VAP). We now show that protrudin associates and colocalizes with VAP-A, an isoform of VAP expressed in the endoplasmic reticulum. Both the interaction between protrudin and VAP-A as well as the induction of process formation by protrudin were markedly inhibited by mutation of the FFAT motif. Furthermore, depletion of VAP-A by RNA interference resulted in mislocalization of protrudin as well as in inhibition of neurite outgrowth induced by nerve growth factor in rat pheochromocytoma PC12 cells. These defects resulting from depletion of endogenous rat VAP-A in PC12 cells were corrected by forced expression of (RNA interference-resistant) human VAP-A but not by VAP-A mutants that have lost the ability to interact with protrudin. These results suggest that VAP-A is an important regulator both of the subcellular localization of protrudin and of its ability to stimulate neurite outgrowth. PMID- 19289471 TI - Protein kinase D mediates mitogenic signaling by Gq-coupled receptors through protein kinase C-independent regulation of activation loop Ser744 and Ser748 phosphorylation. AB - Rapid protein kinase D (PKD) activation and phosphorylation via protein kinase C (PKC) have been extensively documented in many cell types cells stimulated by multiple stimuli. In contrast, little is known about the role and mechanism(s) of a recently identified sustained phase of PKD activation in response to G protein coupled receptor agonists. To elucidate the role of biphasic PKD activation, we used Swiss 3T3 cells because PKD expression in these cells potently enhanced duration of ERK activation and DNA synthesis in response to G(q)-coupled receptor agonists. Cell treatment with the preferential PKC inhibitors GF109203X or Go6983 profoundly inhibited PKD activation induced by bombesin stimulation for <15 min but did not prevent PKD catalytic activation induced by bombesin stimulation for longer times (>60 min). The existence of sequential PKC-dependent and PKC independent PKD activation was demonstrated in 3T3 cells stimulated with various concentrations of bombesin (0.3-10 nm) or with vasopressin, a different G(q) coupled receptor agonist. To gain insight into the mechanisms involved, we determined the phosphorylation state of the activation loop residues Ser(744) and Ser(748). Transphosphorylation targeted Ser(744), whereas autophosphorylation was the predominant mechanism for Ser(748) in cells stimulated with G(q)-coupled receptor agonists. We next determined which phase of PKD activation is responsible for promoting enhanced ERK activation and DNA synthesis in response to G(q)-coupled receptor agonists. We show, for the first time, that the PKC independent phase of PKD activation mediates prolonged ERK signaling and progression to DNA synthesis in response to bombesin or vasopressin through a pathway that requires epidermal growth factor receptor-tyrosine kinase activity. Thus, our results identify a novel mechanism of G(q)-coupled receptor-induced mitogenesis mediated by sustained PKD activation through a PKC-independent pathway. PMID- 19289472 TI - Cyclopiazonic acid is complexed to a divalent metal ion when bound to the sarcoplasmic reticulum Ca2+-ATPase. AB - We have determined the structure of the sarco(endo)plasmic reticulum Ca(2+) ATPase (SERCA) in an E2.P(i)-like form stabilized as a complex with MgF(4)(2-), an ATP analog, adenosine 5'-(beta,gamma-methylene)triphosphate (AMPPCP), and cyclopiazonic acid (CPA). The structure determined at 2.5A resolution leads to a significantly revised model of CPA binding when compared with earlier reports. It shows that a divalent metal ion is required for CPA binding through coordination of the tetramic acid moiety at a characteristic kink of the M1 helix found in all P-type ATPase structures, which is expected to be part of the cytoplasmic cation access pathway. Our model is consistent with the biochemical data on CPA function and provides new measures in structure-based drug design targeting Ca(2+) ATPases, e.g. from pathogens. We also present an extended structural basis of ATP modulation pinpointing key residues at or near the ATP binding site. A structural comparison to the Na(+),K(+)-ATPase reveals that the Phe(93) side chain occupies the equivalent binding pocket of the CPA site in SERCA, suggesting an important role of this residue in stabilization of the potassium-occluded E2 state of Na(+),K(+)-ATPase. PMID- 19289473 TI - There might be more to SPG4! PMID- 19289474 TI - The unsolved puzzle of multiple sclerosis and venous function. PMID- 19289475 TI - Botulinum toxin should be first-line treatment for poststroke spasticity. PMID- 19289476 TI - Diagnosis and new treatment in muscle channelopathies. AB - The skeletal muscle fibre membrane plays a major role in muscle contraction by generating and propagating action potentials, and linking the latter to the release of intracellular calcium stores which triggers mechanical contraction. This function relies on the proper functioning of ion channels. In the last two decades, diseases caused by mutations in muscle ion channel genes have been identified, the so-called muscle channelopathies. Even though the pathophysiology of muscle channelopathies is not completely elucidated, major advances have been made in their understanding, thus linking patient symptoms and neurophysiology with abnormal functioning of the muscle membrane. This has facilitated significant progress both in the diagnosis of these disorders and in the rationale for therapeutic intervention. In this review, we will focus on diagnosis and treatments of muscle channelopathies of relevance to the clinical neurologist. PMID- 19289477 TI - Risk of incident Parkinson's disease and parkinsonism in essential tremor: a population based study. AB - BACKGROUND: A longstanding literature suggests an association between essential tremor (ET) and Parkinson's disease (PD). However, the risk of incident PD has not been quantified in cases of ET compared with controls. OBJECTIVE: To estimate the risk of incident PD in a population based cohort study of 3813 older people (including ET cases and controls) in central Spain. RESULTS: After a median of 3.3 years, 12 (5.8%) of 207 ET cases developed parkinsonism compared with 56 (1.6%) of 3606 controls (adjusted relative risk (RR) 3.47, 95% CI 1.82 to 6.59; p<0.001). Six (3.0%) of 201 ET cases developed incident PD versus 24 (0.7%) of 3574 controls (adjusted RR 4.27, 95% CI 1.72 to 10.61; p = 0.002). CONCLUSIONS: Patients with ET were four times more likely than controls to develop incident PD during prospective follow-up. These data confirm and begin to quantify the link between these two diseases. PMID- 19289478 TI - Filamin A mutation, a common cause for periventricular heterotopia, aneurysms and cardiac defects. AB - Filamin A is an important gene involved in the development of the brain, heart, connective tissue and blood vessels. A case is presented illustrating the challenge in recognising patients with filamin A mutations. The patient, a 71 year-old woman, was known to have heart valve disease and bilateral periventricular nodular heterotopia when she died of a subarachnoid haemorrhage. Autopsy showed typical cerebral bilateral periventricular heterotopia and vascular abnormalities. Postmortally, the diagnosis of a filamin A mutation was confirmed. Recognition during life may prevent cardiovascular problems and provide possibilities for genetic counselling. PMID- 19289479 TI - Disrupted intrinsic functional connectivity in the vegetative state. AB - It is debatable as to whether the spontaneous blood-oxygen-level dependent fluctuations that are observed in the resting brain in turn reflect consciously directed mental activity or, alternatively, constitute an intrinsic property of functional brain organisation persisting in the absence of consciousness. This report shows for the first time, in three patients, that the persistent vegetative state (PVS) is marked by a dysfunctional default mode network, with decreased connectivity in several brain regions, including the dorsolateral prefrontal cortex and anterior cingulated cortex, especially in the right hemisphere. This finding supports the view that the resting state is involved in self-consciousness, and that the right-hemisphere default state may play a major role in conscious processes. It is speculated that the default state may act as a surrogate marker of PVS with awareness contents and, therefore, could replace a more complex activation paradigm. PMID- 19289480 TI - Pattern and progression of white-matter changes in a case of posterior cortical atrophy using diffusion tensor imaging. AB - BACKGROUND: The progression of white-matter changes in a case of posterior cortical atrophy (PCA) was examined over a period of 15 months using diffusion tensor imaging (DTI) and the association with neuropsychological variables was studied. PATIENT AND METHODS: A PCA patient was observed over a period of 15 months. DTI and volumetric magnetic resonance imaging were obtained at visit 1 and 15 months later. Fractional anisotropy (FA) and volumetric changes were compared with findings in a typical case of Alzheimer disease (AD) and in 65 healthy volunteers, and the association of neuropsychological deficits with these changes was studied. RESULTS: Reduction in FA was focused on the occipital lobe in the early stages of PCA. During the 15-month period, the FA values of the PCA patient tended to align with the FA ratios of the AD patient, with a more pronounced FA reduction in the parietal lobes, as opposed to a stable FA level in the occipital lobe. In addition to the DTI changes, clinical and neuropsychological symptoms deteriorated further. Brain volumes (grey matter, white matter and total normalised brain volume) of the PCA patient were substantially decreased compared with the control group, but loss of tissue volumes showed only marginal progression between visit 1 and 2. CONCLUSIONS: The findings suggest that PCA starts as distinct clinical syndrome but in its later course might turn into a final pathway shared with AD. DTI might be helpful in detecting changes in cerebral white matter during disease progression in PCA patients. PMID- 19289481 TI - Risk indicators for development of headache during dipyridamole treatment after cerebral ischaemia of arterial origin. AB - A considerable proportion of patients discontinue dipyridamole therapy because of headache. Risk indicators for the development of dipyridamole induced headache were identified by means of an exploratory analysis of data from the European/Australasian Stroke Prevention in Reversible Ischaemia Trial (ESPRIT) and the Second European Stroke Prevention Study (ESPS 2). In ESPRIT, dipyridamole induced headache was significantly associated with female sex, absence of hypertension and non-smoking (area under the receiver operator characteristic (ROC) curve 0.63 (95% CI 0.58 to 0.68)) and in ESPS 2 with female sex and absence of ischaemic lesions on imaging (area under the ROC curve 0.64 (95% CI 0.59 to 0.69)). PMID- 19289482 TI - Posterior fossa abnormalities in hereditary spastic paraparesis with spastin mutations. AB - BACKGROUND: Hereditary spastic paraparesis (HPS) linked to mutations in the spastin gene (SPG4) is considered to be a pure form of spastic hereditary paraparesis. However, in this disease also other signs of central nervous system involvement are frequently found. METHODS: Clinical, genetical and neuroradiological investigations were carried out in a large family with autosomal dominant spastic paraparesis and in a sporadic case with spastic paraparesis. RESULTS: Additional clinical and molecular data are provided, studying other members of the same pedigree, as already described, with a five base deletion in exon 9 of the SPG4 gene (1215-1219delTATAA) whose members show MRI anomalies that fall within the Dandy-Walker continuum. Furthermore, an unrelated female patient with hypoplasia of the cerebellar vermis is indicated, carrying a de novo previously reported mutation of the SPG4 gene (c.1741C>T p.R581X). CONCLUSIONS: Spastin may play an important role in the development of the central nervous system and in particular in the development of the structures of posterior fossa. PMID- 19289483 TI - Chronic ophthalmoparesis in limb girdle muscular dystrophy 1C. PMID- 19289484 TI - Definite multiple system atrophy in a German family. PMID- 19289485 TI - Novel epsilon subunit mutation of the muscle acetylcholine receptor causing a slow-channel congenital myasthenic syndrome. PMID- 19289487 TI - Minimal detectable change and clinically important difference of the Wolf Motor Function Test in stroke patients. AB - OBJECTIVES: This study aimed to establish the minimal detectable change (MDC) and clinically important differences (CID) of the Wolf Motor Function Test (WMFT) in patients with stroke, and to assess the proportions of patients' change scores exceeding the MDC and CID after stroke rehabilitation. METHODS: A total of 57 patients received 1 of the 3 treatments for 3 weeks and underwent clinical assessments before and after treatment. The MDC, at 90% confidence (MDC( 90)), was calculated from the standard error of measurement to indicate a real change for individual patients. Anchor-based and distribution-based approaches were used to triangulate the values of minimal CID. The percentages of patients exceeding the MDC and minimal CID were also examined. RESULTS: The MDC(90) of the WMFT was 4.36 for the performance time (WMFT time) and 0.37 for the functional ability scale (WMFT FAS). The minimal CID ranged from 1.5 to 2 seconds on the WMFT time and from 0.2 to 0.4 points on the WMFT FAS. The MDC and CID proportions ranged from 14% to 30% on the WMFT time and from 39% to 65% on the WMFT FAS, respectively. CONCLUSIONS: The change score of an individual patient has to reach 4.36 and 0.37 on the WMFT time and WMFT FAS to indicate a real change. The mean change scores of a stroke group on the WMFT time and WMFT FAS should achieve 1.5 to 2 seconds and 0.2 to 0.4 points to be regarded as clinically important changes. Furthermore, the WMFT FAS may be more responsive than the WMFT time based on the results of proportions exceeding the threshold criteria. PMID- 19289488 TI - Immunotherapy of acute myeloid leukemia: current approaches. AB - Following standard therapy that consists of chemotherapy with or without stem cell transplantation, both relapsed and refractory disease shorten the survival of acute myeloid leukemia (AML) patients. Therefore, additional treatment options are urgently needed, especially to fight residual AML cells. The identification of leukemia-associated antigens and the observation that administration of allogeneic T cells can mediate a graft-versus-leukemia effect paved the way to the development of active and passive immunotherapy strategies, respectively. The aim of these strategies is the eradication of AML cells by the immune system. In this review, an overview is provided of both active and passive immunotherapy strategies that are under investigation or in use for the treatment of AML. For each strategy, a critical view on the state of the art is given and future perspectives are discussed. PMID- 19289489 TI - Risks and benefits of whole body vibration training in older people. PMID- 19289490 TI - REV3L confers chemoresistance to cisplatin in human gliomas: the potential of its RNAi for synergistic therapy. AB - The REV3L gene, encoding the catalytic subunit of human polymerase zeta, plays a significant role in the cytotoxicity, mutagenicity, and chemoresistance of certain tumors. However, the role of REV3L in regulating the sensitivity of glioma cells to chemotherapy remains unknown. In this study, we investigated the expression of the REV3L gene in 10 normal brain specimens and 30 human glioma specimens and examined the value of REV3L as a potential modulator of cellular response to various DNA-damaging agents. Reverse transcriptase PCR/real-time PCR analysis revealed that REV3L was overexpressed in human gliomas compared with normal brain tissues. A glioma cell model with stable overexpression of REV3L was used to probe the role of REV3L in cisplatin treatment; upregulation of REV3L markedly attenuated cisplatin-induced apoptosis of the mitochondrial apoptotic pathway. We therefore assessed the REV3L-targeted treatment modality that combines suppression of REV3L expression using RNA interference (RNAi) with the cytotoxic effects of DNA-damaging agents. Downregulation of REV3L expression significantly enhanced the sensitivity of glioma cells to cisplatin, as evidenced by the increased apoptosis rate and marked alterations in the anti-apoptotic proteins B-cell lymphoma 2 (Bcl-2) and B-cell lymphoma-extra large (Bcl-xl) and proapoptotic Bcl-2-associated x protein (Bax) expression levels, and reduced mutation frequencies in surviving glioma cells. These results suggest that REV3L may potentially contribute to gliomagenesis and play a crucial role in regulating cellular response to the DNA cross-linking agent cisplatin. Our findings indicate that RNAi targeting REV3L combined with chemotherapy has synergistic therapeutic effects on glioma cells, which warrants further investigation as an effective novel therapeutic regimen for patients with this malignancy. PMID- 19289491 TI - Phase I trial of temozolomide plus O6-benzylguanine 5-day regimen with recurrent malignant glioma. AB - This phase I clinical trial conducted with patients who had recurrent or progressive malignant glioma (MG) was designed to determine the maximum tolerated dose (MTD) and toxicity of three different 5-day dosing regimens of temozolomide (TMZ) in combination with O(6)-benzylguanine (O(6)-BG). Both TMZ and O(6)-BG were administered on days 1-5 of a 28-day treatment cycle. A bolus infusion of O(6)-BG was administered at 120 mg/m(2) over 1 h on days 1, 3, and 5, along with a continuous infusion of O(6)-BG at 30 mg/m(2)/day. TMZ was administered at the end of the first bolus infusion of O(6)-BG and then every 24 h for 5 days during the continuous infusion of O(6)-BG. Patients were accrued to one of three 5-day dosing regimens of TMZ. Twenty-nine patients were enrolled into this study. The dose-limiting toxicities (DLTs) were grade 4 neutropenia, leukopenia, and thrombocytopenia. The MTD for TMZ for the three different 5-day dosing schedules was determined as follows: schedule 1, 200 mg/m(2) on day 1 and 50 mg/m(2)/day on days 2-5; schedule 2, 50 mg/m(2)/day on days 1-5; and schedule 3, 50 mg/m(2)/day on days 1-5 while receiving pegfilgrastim. Thus, the 5-day TMZ dosing schedule that maximized the total dose of TMZ when combined with O(6)-BG was schedule 1. This study provides the foundation for a phase II trial of O(6)-BG in combination with a 5-day dosing schedule of TMZ in TMZ-resistant MG. PMID- 19289492 TI - Pseudoprogression in boron neutron capture therapy for malignant gliomas and meningiomas. AB - Pseudoprogression has been recognized and widely accepted in the treatment of malignant gliomas, as transient increases in the volume of the enhanced area just after chemoradiotherapy, especially using temozolomide. We experienced a similar phenomenon in the treatment of malignant gliomas and meningiomas using boron neutron capture therapy (BNCT), a cell-selective form of particle radiation. Here, we introduce representative cases and analyze the pathogenesis. Fifty-two cases of malignant glioma and 13 cases of malignant meningioma who were treated by BNCT were reviewed retrospectively mainly via MR images. Eleven of 52 malignant gliomas and 3 of 13 malignant meningiomas showed transient increases of enhanced volume in MR images within 3 months after BNCT. Among these cases, five patients with glioma underwent surgery because of suspicion of relapse. In histology, most of the specimens showed necrosis with small amounts of residual tumor cells. Ki-67 labeling showed decreased positivity compared with previous samples from the individuals. Fluoride-labeled boronophenylalanine PET was applied in four and two cases of malignant gliomas and meningiomas, respectively, at the time of transient increase of lesions. These PET scans showed decreased lesion:normal brain ratios in all cases compared with scans obtained prior to BNCT. With or without surgery, all lesions were decreased or stable in size during observation. Transient increases in enhanced volume in malignant gliomas and meningiomas immediately after BNCT seemed to be pseudoprogression. This pathogenesis was considered as treatment-related intratumoral necrosis in the subacute phase after BNCT. PMID- 19289493 TI - Leptin deficiency and beta-cell dysfunction underlie type 2 diabetes in compound Akt knockout mice. AB - Phenotypic analyses of mice null for the individual Akt isoforms suggested that they are functionally distinct and that only Akt2 plays a role in diabetes. We show here that Akt isoforms play compensatory and complementary roles in glucose homeostasis and diabetes. Insulin resistance in Akt2(-/-) mice was inhibited by haplodeficiency of Pten, suggesting that other Akt isoforms can compensate for Akt2 function. Haplodeficiency of Akt1 in Akt2(-/-) mice, however, converts prediabetes to overt type 2 diabetes, which is also reversed by haplodeficiency of Pten. Akt3 does not appear to contribute significantly to diabetes. Overt type 2 diabetes in Akt1(+/-) Akt2(-/-) mice is manifested by hyperglycemia due to beta cell dysfunction combined with impaired glucose homeostasis due to markedly decreased leptin levels. Restoring leptin levels was sufficient to restore normal blood glucose and insulin levels in Akt1(+/-) Akt2(-/-) and Akt2(-/-) mice, suggesting that leptin-deficiency is the predominant cause of diabetes in these mice. These results uncover a new mechanism linking Akt to diabetes, provide a therapeutic strategy, and show that diabetes induced as a consequence of cancer therapy, via Akt inhibition, could be reversed by leptin therapy. PMID- 19289494 TI - A mouse PRMT1 null allele defines an essential role for arginine methylation in genome maintenance and cell proliferation. AB - Protein arginine methyltransferase 1 (PRMT1) is the major enzyme that generates monomethylarginine and asymmetrical dimethylarginine. We report here a conditional null allele of PRMT1 in mice and that the loss of PRMT1 expression leads to embryonic lethality. Using the Cre/lox-conditional system, we show that the loss of PRMT1 in mouse embryonic fibroblasts (MEFs) leads to the loss of arginine methylation of substrates harboring a glycine-arginine rich motif, including Sam68 and MRE11. The loss of PRMT1 in MEFs leads to spontaneous DNA damage, cell cycle progression delay, checkpoint defects, aneuploidy, and polyploidy. We show using a 4-hydroxytamoxifen-inducible Cre that the loss of PRMT1 in MEFs leads to a higher incidence of chromosome losses, gains, structural rearrangements, and polyploidy, as documented by spectral karyotyping. Using PRMT1 small interfering RNA in U2OS cells, we further show that PRMT1-deficient cells are hypersensitive to the DNA damaging agent etoposide and exhibit a defect in the recruitment of the homologous recombination RAD51 recombinase to DNA damage foci. Taken together, these data show that PRMT1 is required for genome integrity and cell proliferation. Our findings also suggest that arginine methylation by PRMT1 is a key posttranslational modification in the DNA damage response pathway in proliferating mammalian cells. PMID- 19289495 TI - Trophoblast stem cell maintenance by fibroblast growth factor 4 requires MEKK4 activation of Jun N-terminal kinase. AB - Trophoblast differentiation during placentation involves an epithelial mesenchymal transition (EMT) with loss of E-cadherin and gain of trophoblast invasiveness. Mice harboring a point mutation that renders inactive the mitogen activated protein kinase kinase kinase MEKK4 exhibit dysregulated placental development with increased trophoblast invasion. Isolated MEKK4 kinase-inactive trophoblast stem (TS) cells cultured under undifferentiating, self-renewing conditions in the presence of fibroblast growth factor 4 (FGF4) display increased expression of Slug, Twist, and matrix metalloproteinase 2 (MMP2), loss of E cadherin, and hyperinvasion of extracellular matrix, each a hallmark of EMT. MEKK4 kinase-inactive TS cells show a preferential differentiation to Tpbp alpha- and Gcm1-positive trophoblasts, which are indicative of spongiotrophoblast and syncytiotrophoblast differentiation, respectively. FGF4-stimulated Jun N-terminal kinase (JNK) and p38 activity is markedly reduced in MEKK4 kinase-inactive TS cells. Chemical inhibition of JNK in wild-type TS cells induced a similar EMT response as loss of MEKK4 kinase activity, including inhibition of E-cadherin expression and increased expression of Slug, MMP2, Tpbp alpha, and Gcm1. Chromatin immunoprecipitation analyses revealed changes in AP-1 composition with increased Fra-2 and decreased Fra-1 and JunB binding to the regulatory regions of Gcm1 and MMP2 genes in MEKK4 kinase-inactive TS cells. Our results define MEKK4 as a signaling hub for FGF4 activation of JNK that is required for maintenance of TS cells in an undifferentiated state. PMID- 19289496 TI - Pak4, a novel Gab1 binding partner, modulates cell migration and invasion by the Met receptor. AB - Hepatocyte growth factor (HGF), the ligand for the Met receptor tyrosine kinase, induces epithelial cell dispersal, invasion, and morphogenesis, events that require remodeling of the actin cytoskeleton. The scaffold protein Gab1 is essential for these biological responses downstream from Met. We have identified p21-activated kinase 4 (Pak4) as a novel Gab1-interacting protein. We show that in response to HGF, Gab1 and Pak4 associate and colocalize at the cell periphery within lamellipodia. The association between Pak4 and Gab1 is dependent on Gab1 phosphorylation but independent of Pak4 kinase activity. The interaction is mediated through a region in Gab1, which displays no homology to known Gab1 interaction motifs and through the guanine exchange factor-interacting domain of Pak4. In response to HGF, Gab1 and Pak4 synergize to enhance epithelial cell dispersal, migration, and invasion, whereas knockdown of Pak4 attenuates these responses. A Gab1 mutant unable to recruit Pak4 fails to promote epithelial cell dispersal and an invasive morphogenic program in response to HGF, demonstrating a physiological requirement for Gab1-Pak4 association. These data demonstrate a novel association between Gab1 and Pak4 and identify Pak4 as a key integrator of cell migration and invasive growth downstream from the Met receptor. PMID- 19289497 TI - Interferon-dependent engagement of eukaryotic initiation factor 4B via S6 kinase (S6K)- and ribosomal protein S6K-mediated signals. AB - Although the roles of Jak-Stat pathways in type I and II interferon (IFN) dependent transcriptional regulation are well established, the precise mechanisms of mRNA translation for IFN-sensitive genes remain to be defined. We examined the effects of IFNs on the phosphorylation/activation of eukaryotic translation initiation factor 4B (eIF4B). Our data show that eIF4B is phosphorylated on Ser422 during treatment of sensitive cells with alpha IFN (IFN-alpha) or IFN gamma. Such phosphorylation is regulated, in a cell type-specific manner, by either the p70 S6 kinase (S6K) or the p90 ribosomal protein S6K (RSK) and results in enhanced interaction of the protein with eIF3A (p170/eIF3A) and increased associated ATPase activity. Our data also demonstrate that IFN-inducible eIF4B activity and IFN-stimulated gene 15 protein (ISG15) or IFN-gamma-inducible chemokine CXCL-10 protein expression are diminished in S6k1/S6k2 double-knockout mouse embryonic fibroblasts. In addition, IFN-alpha-inducible ISG15 protein expression is blocked by eIF4B or eIF3A knockdown, establishing a requirement for these proteins in mRNA translation/protein expression by IFNs. Importantly, the generation of IFN-dependent growth inhibitory effects on primitive leukemic progenitors is dependent on activation of the S6K/eIF4B or RSK/eIF4B pathway. Taken together, our findings establish critical roles for S6K and RSK in the induction of IFN-dependent biological effects and define a key regulatory role for eIF4B as a common mediator and integrator of IFN-generated signals from these kinases. PMID- 19289498 TI - Regulation of p53 by TopBP1: a potential mechanism for p53 inactivation in cancer. AB - Proper control of the G(1)/S checkpoint is essential for normal proliferation. The activity of p53 must be kept at a very low level under unstressed conditions to allow growth. Here we provide evidence supporting a crucial role for TopBP1 in actively repressing p53. Depletion of TopBP1 upregulates p53 target genes involved in cell cycle arrest and apoptosis and enhances DNA damage-induced apoptosis. The regulation is mediated by an interaction between the seventh and eighth BRCT domains of TopBP1 and the DNA-binding domain of p53, leading to inhibition of p53 promoter binding activity. Importantly, TopBP1 overexpression is found in 46 of 79 primary breast cancer tissues and is associated with high tumor grade and shorter patient survival time. Overexpression of TopBP1 to a level comparable to that seen in breast tumors leads to inhibition of p53 target gene expression and DNA damage-induced apoptosis and G(1) arrest. Thus, a physiological level of TopBP1 is essential for normal G(1)/S transition, but a pathological level of TopBP1 in cancer may perturb p53 function and contribute to an aggressive tumor behavior. PMID- 19289499 TI - p65/RelA modulates BECN1 transcription and autophagy. AB - Recently, autophagy has emerged as a critical process in the control of T-cell homeostasis. Given the pivotal role of NF-kappaB in the signaling events of T cells, we have analyzed and unveiled a conserved NF-kappaB binding site in the promoter of the murine and human BECN1 autophagic gene (Atg6). Accordingly, we demonstrate that the NF-kappaB family member p65/RelA upregulates BECN1 mRNA and protein levels in different cellular systems. Moreover, p65-mediated upregulation of BECN1 is coupled to increased autophagy. The newly identified kappaB site in the BECN1 promoter specifically interacts with p65 both in vitro and in living Jurkat cells upon phorbol myristate acetate (PMA)-ionomycin stimulation, where p65 induction is coupled to BECN1 upregulation and autophagy induction. Finally, anti-CD3- and PMA-ionomycin-mediated activation of T-cell receptor signaling in peripheral T cells from lymph nodes of healthy mice results in an upregulation of BECN1 expression that can be blocked by the NF-kappaB inhibitor BAY 11-7082. Altogether, these data suggest that autophagy could represent a novel route modulated by p65 to regulate cell survival and control T-cell homeostasis. PMID- 19289500 TI - Analysis of nitric oxide-stabilized mRNAs in human fibroblasts reveals HuR dependent heme oxygenase 1 upregulation. AB - We previously observed that nitric oxide (NO) exposure increases the stability of mRNAs encoding heme oxygenase 1 (HO-1) and TIEG-1 in human and mouse fibroblasts. Here, we have used microarrays to look broadly for changes in mRNA stability in response to NO treatment. Using human IMR-90 and mouse NIH 3T3 fibroblasts treated with actinomycin D to block de novo transcription, microarray analysis suggested that the stability of the majority of mRNAs was unaffected. Among the mRNAs that were stabilized by NO treatment, seven transcripts were found in both IMR-90 and NIH 3T3 cells (CHIC2, GADD45B, HO-1, PTGS2, RGS2, TIEG, and ID3) and were chosen for further analysis. All seven mRNAs showed at least one hit of a signature motif for the stabilizing RNA-binding protein (RBP) HuR; accordingly, ribonucleoprotein immunoprecipitation analysis revealed that all seven mRNAs associated with HuR. In keeping with a functional role of HuR in the response to NO, a measurable fraction of HuR increased in the cytoplasm following NO treatment. However, among the seven transcripts, only HO-1 mRNA showed a robust increase in the level of its association with HuR following NO treatment. In turn, HO-1 mRNA and protein levels were significantly reduced when HuR levels were silenced in IMR-90 cells, and they were elevated when HuR was overexpressed. In sum, our results indicate that NO stabilizes mRNA subsets in fibroblasts, identify HuR as an RBP implicated in the NO response, reveal that HuR alone is insufficient for stabilizing several mRNAs by NO, and show that HO-1 induction by NO is regulated by HuR. PMID- 19289501 TI - Hnf1alpha (MODY3) controls tissue-specific transcriptional programs and exerts opposed effects on cell growth in pancreatic islets and liver. AB - Heterozygous HNF1A mutations cause pancreatic-islet beta-cell dysfunction and monogenic diabetes (MODY3). Hnf1alpha is known to regulate numerous hepatic genes, yet knowledge of its function in pancreatic islets is more limited. We now show that Hnf1a deficiency in mice leads to highly tissue-specific changes in the expression of genes involved in key functions of both islets and liver. To gain insights into the mechanisms of tissue-specific Hnf1alpha regulation, we integrated expression studies of Hnf1a-deficient mice with identification of direct Hnf1alpha targets. We demonstrate that Hnf1alpha can bind in a tissue selective manner to genes that are expressed only in liver or islets. We also show that Hnf1alpha is essential only for the transcription of a minor fraction of its direct-target genes. Even among genes that were expressed in both liver and islets, the subset of targets showing functional dependence on Hnf1alpha was highly tissue specific. This was partly explained by the compensatory occupancy by the paralog Hnf1beta at selected genes in Hnf1a-deficient liver. In keeping with these findings, the biological consequences of Hnf1a deficiency were markedly different in islets and liver. Notably, Hnf1a deficiency led to impaired large-T-antigen-induced growth and oncogenesis in beta cells yet enhanced proliferation in hepatocytes. Collectively, these findings show that Hnf1alpha governs broad, highly tissue-specific genetic programs in pancreatic islets and liver and reveal key consequences of Hnf1a deficiency relevant to the pathophysiology of monogenic diabetes. PMID- 19289502 TI - Spi-1 and Fli-1 directly activate common target genes involved in ribosome biogenesis in Friend erythroleukemic cells. AB - Spi-1 and Fli-1 are ETS transcription factors recurrently deregulated in mouse erythroleukemia induced by Friend viruses. Since they share the same core DNA binding site, we investigated whether they may contribute to erythroleukemia by common mechanisms. Using inducible knockdown, we demonstrated that Fli-1 contributes to proliferation, survival, and differentiation arrest of erythroleukemic cells harboring an activated fli-1 locus. Similarly, we used inducible Fli-1 knockdown and either hexamethylenebisacetamide (HMBA)- or small interfering RNA-mediated Spi-1 knockdown to investigate their respective contributions in erythroleukemic cells harboring an activated spi-1 locus. In these cells, simple or double knockdown of both Spi-1 and Fli-1 additively contributed to induce proliferation arrest and differentiation. Transcriptome profiling revealed that virtually all transcripts affected by both Fli-1 knockdown and HMBA are affected in an additive manner. Among these additively downregulated transcripts, more than 20% encode proteins involved in ribosome biogenesis, and conserved ETS binding sites are present in their gene promoters. Through chromatin immunoprecipitation, we demonstrated the association of Spi-1 and Fli-1 on these promoters in Friend erythroleukemic cells. These data lead us to propose that the oncogenicity of Spi-1, Fli-1, and possibly other ETS transcription factors may involve their ability to stimulate ribosome biogenesis. PMID- 19289503 TI - Limiting the extent of the RDN1 heterochromatin domain by a silencing barrier and Sir2 protein levels in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, transcriptional silencing occurs at the cryptic mating-type loci (HML and HMR), telomeres, and ribosomal DNA (rDNA; RDN1). Silencing in the rDNA is unusual in that polymerase II (Pol II) promoters within RDN1 are repressed by Sir2 but not Sir3 or Sir4. rDNA silencing unidirectionally spreads leftward, but the mechanism of limiting its spreading is unclear. We searched for silencing barriers flanking the left end of RDN1 by using an established assay for detecting barriers to HMR silencing. Unexpectedly, the unique sequence immediately adjacent to RDN1, which overlaps a prominent cohesin binding site (CARL2), did not have appreciable barrier activity. Instead, a fragment located 2.4 kb to the left, containing a tRNA(Gln) gene and the Ty1 long terminal repeat, had robust barrier activity. The barrier activity was dependent on Pol III transcription of tRNA(Gln), the cohesin protein Smc1, and the SAS1 and Gcn5 histone acetyltransferases. The location of the barrier correlates with the detectable limit of rDNA silencing when SIR2 is overexpressed, where it blocks the spreading of rDNA heterochromatin. We propose a model in which normal Sir2 activity results in termination of silencing near the physical rDNA boundary, while tRNA(Gln) blocks silencing from spreading too far when nucleolar Sir2 pools become elevated. PMID- 19289504 TI - Polo-like kinase 1 depletion induces DNA damage in early S prior to caspase activation. AB - Polo-like kinase 1 (Plk1) plays several roles in mitosis, and it has been suggested to have a role in tumorigenesis. We have previously reported that Plk1 depletion results in cell death in cancer cells, whereas normal cells survive similar depletion. However, Plk1 depletion together with p53 depletion induces cell death in normal cells as well. This communication presents evidence on the sequence of events that leads to cell death in cancer cells. DNA damage is detected at the first S phase following Plk1 depletion and is more severe in Plk1 depleted p53-null cancer cells. As a consequence of Plk1 depletion using lentivirus-based small interfering RNA techniques, prereplicative complex (pre RC) formation is disrupted at the G(1)/S transition, and DNA synthesis is reduced during S phase of the first cycle after depletion. The levels of geminin, an inhibitor of DNA pre-RC, and Emi1, an inhibitor of anaphase-promoting complex/cyclosome, are elevated in Plk1-depleted cells. The rate of cell cycling is slower in Plk1-depleted cells than in control cells when synchronized by serum starvation. Plk1 depletion results in disrupted DNA pre-RC formation, reduced DNA synthesis, and DNA damage before cells display severe mitotic catastrophe or apoptosis. Our data suggest that Plk1 is required for cell cycle progression not only in mitosis but also for DNA synthesis, maintenance of DNA integrity, and prevention of cell death. PMID- 19289505 TI - Multivalent binding of the ETO corepressor to E proteins facilitates dual repression controls targeting chromatin and the basal transcription machinery. AB - E proteins are a family of helix-loop-helix transcription factors that play important roles in cell differentiation and homeostasis. They contain at least two activation domains, AD1 and AD2. ETO family proteins and the leukemogenic AML1-ETO fusion protein are corepressors of E proteins. It is thought that ETO represses E-protein activity by interacting with AD1, which competes away p300/CBP histone acetyltransferases. Here we report that E proteins contain another conserved ETO-interacting region, termed DES, and that differential associations with AD1 and DES allow ETO to repress transcription through both chromatin-dependent and chromatin-independent mechanisms. At the chromatin level, AD1 and AD2 cooperatively recruit p300. ETO interacts with AD1 to abolish p300 recruitment and to allow HDAC-dependent silencing. At the post-chromatin remodeling level, binding to DES enables ETO to directly inhibit activation of the basal transcription machinery. This novel repression mechanism is conserved in ETO family proteins and in the AML1-ETO fusion protein. In addition, the repression capacity exerted by each mechanism is differentially modulated by cross talk among various ETO domains and the AML1 domain of AML1-ETO. In particular, the oligomerization domain of ETO plays a major role in targeting ETO to the DES region and independently potentiates the TAFH domain-mediated AD1 interaction. The ability to exert repression at different levels not only may allow these corepressors to impose robust inhibition of signal-independent transcription but may also allow a rapid response to signals. In addition, our newly defined domain interactions and their interplays have important implications in effectively targeting both E-protein fusion proteins and AML1-ETO found in cancers. PMID- 19289506 TI - Molecular Darwinian evolution of virulence in Yersinia pestis. PMID- 19289507 TI - Mechanisms of the hepatic acute-phase response during bacterial pneumonia. AB - The acute-phase response is characterized by increased circulating levels of acute-phase proteins (APPs) generated by the liver. During bacterial pneumonia, APPs correlate with the severity of disease, serve as biomarkers, and are functionally significant. The kinetics and regulatory mechanisms of APP induction in the liver during lung infection have yet to be defined. Here we show that APP mRNA transcription is induced in the livers of mice whose lungs are infected with either Escherichia coli or Streptococcus pneumoniae, and that in both cases this induction occurs in tandem with activation in the liver of the transcription factors signal transducer and activator of transcription 3 (STAT3) and NF-kappaB RelA. Interleukin-6 (IL-6) deficiency inhibited the activation of STAT3 and the induction of select APPs in the livers of pneumonic mice. Furthermore, liver RelA activation and APP induction were reduced for mice lacking all signaling receptors for tumor necrosis factor alpha and IL-1. In a murine hepatocyte cell line, knockdown of either STAT3 or RelA by small interfering RNA inhibited cytokine induction of the APP serum amyloid A-1, demonstrating that both transcription factors were independently essential for the expression of this gene. These data suggest that during pneumonia caused by gram-negative or gram positive bacteria, the expression of APPs in the liver depends on STAT3 activation by IL-6 and on RelA activation by early-response cytokines. These signaling axes may be critical for integrating systemic responses to local infection, balancing antibacterial host defenses and inflammatory injury during acute bacterial pneumonia. PMID- 19289508 TI - Identification of cross-reactive epitopes on the conserved 47-kilodalton antigen of Orientia tsutsugamushi and human serine protease. AB - Orientia tsutsugamushi is the causative agent of scrub typhus. One of the protein antigens of this species, the conserved 47-kDa protein (HtrA), has been shown to induce an antibody response in patients and can provide protective immunity against live challenge by Orientia in mice. Pepscan experiments identified many peptide epitope clusters in different parts of this protein. The majority of the most reactive epitopes are located at the C terminus of the protein (from amino acid 333 to amino acid 430). Protein sequence analysis revealed that the 47-kDa protein contains a trypsin domain and has sequence homology to human serine protease HtrA1 (hHtrA1). As the 47-kDa protein is a potential vaccine candidate and its ability to induce autoimmunity is a concern, the reactivity of scrub typhus patient sera with purified recombinant 47-kDa and hHtrA1 proteins was tested. A significant percentage (>20%) of scrub typhus patient sera reacted strongly with recombinant hHTRA1 and two of the antigenic polypeptide epitopes in hHtrA1. These findings suggest that the safety of the full-length 47-kDa antigen as a vaccine candidate is a significant issue due to its cross-reactivity with a human protein, which may also contribute to autoimmune responses or enhanced pathology in some scrub typhus patients. PMID- 19289509 TI - Yersinia pestis can reside in autophagosomes and avoid xenophagy in murine macrophages by preventing vacuole acidification. AB - Yersinia pestis survives and replicates in phagosomes of murine macrophages. Previous studies demonstrated that Y. pestis-containing vacuoles (YCVs) acquire markers of late endosomes or lysosomes in naive macrophages and that this bacterium can survive in macrophages activated with the cytokine gamma interferon. An autophagic process known as xenophagy, which destroys pathogens in acidic autophagolysosomes, can occur in naive macrophages and is upregulated in activated macrophages. Studies were undertaken here to investigate the mechanism of Y. pestis survival in phagosomes of naive and activated macrophages and to determine if the pathogen avoids or co-opts autophagy. Colocalization of the YCV with markers of autophagosomes or acidic lysosomes and the pH of the YCV were determined by microscopic imaging of infected macrophages. Some YCVs contained double membranes characteristic of autophagosomes, as determined by electron microscopy. Fluorescence microscopy showed that approximately 40% of YCVs colocalized with green fluorescent protein (GFP)-LC3, a marker of autophagic membranes, and that YCVs failed to acidify below pH 7 in naive macrophages. Replication of Y. pestis in naive macrophages caused accumulation of LC3-II, as determined by immunoblotting. While activation of infected macrophages increased LC3-II accumulation, it decreased the percentage of GFP-LC3-positive YCVs (approximately 30%). A viable count assay showed that Y. pestis survived equally well in macrophages proficient for autophagy and macrophages rendered deficient for this process by Cre-mediated deletion of ATG5, revealing that this pathogen does not require autophagy for intracellular replication. We conclude that although YCVs can acquire an autophagic membrane and accumulate LC3-II, the pathogen avoids xenophagy by preventing vacuole acidification. PMID- 19289510 TI - Designing therapies against experimental visceral leishmaniasis by modulating the membrane fluidity of antigen-presenting cells. AB - The membrane fluidity of antigen-presenting cells (APCs) has a significant bearing on T-cell-stimulating ability and is dependent on the cholesterol content of the membrane. The relationship, if any, between membrane fluidity and defective cell-mediated immunity in visceral leishmaniasis has been investigated. Systemic administration of cholesterol by liposome delivery (cholesterol liposomes) in Leishmania donovani-infected hamsters was found to cure the infection. Splenic macrophages as a prototype of APCs in infected hamsters had decreased membrane cholesterol and an inability to drive T cells, which was corrected by cholesterol liposome treatment. The effect was cholesterol specific because liposomes made up of the analogue 4-cholesten-3-one provided almost no protection. Infection led to increases in interleukin-10 (IL-10), transforming growth factor beta, and IL-4 signals and concomitant decreases in gamma interferon (IFN-gamma), tumor necrosis factor alpha, and inducible NO synthase signals, which reverted upon cholesterol liposome treatment. The antileishmanial T-cell repertoire, whose expansion appeared to be associated with protection, was presumably type Th1, as shown by enhanced IFN-gamma signals and the predominance of the immunoglobulin G2 isotype. The protected group produced significantly more reactive oxygen species and NO than the infected groups, which culminated in killing of L. donovani parasites. Therefore, cholesterol liposome treatment may be yet another simple strategy to enhance the cell-mediated immune response to L. donovani infection. To our knowledge, this is the first report on the therapeutic effect of cholesterol liposomes in any form of the disease. PMID- 19289511 TI - Genetics and virulence association of the Shigella flexneri sit iron transport system. AB - The sit-encoded iron transport system is present within pathogenicity islands in all Shigella spp. and some pathogenic Escherichia coli strains. The islands contain numerous insertion elements and sequences with homology to bacteriophage genes. The Shigella flexneri sit genes can be lost as a result of deletion within the island. The formation of deletions was dependent upon RecA and occurred at relatively high frequency. This suggests that the sit region is inherently unstable, yet sit genes are maintained in all of the clinical isolates tested. Characterization of the sitABCD genes in S. flexneri indicates that they encode a ferrous iron transport system, although the genes are induced aerobically. The sit genes provide a competitive advantage to S. flexneri growing within epithelial cells, and a sitA mutant is outcompeted by the wild type in cultured epithelial cells. The Sit system is also required for virulence in a mouse lung model. The sitA mutant was able to infect the mice and induce a protective immune response but was avirulent compared to its wild-type parent strain. PMID- 19289512 TI - Lex2B, a phase-variable glycosyltransferase, adds either a glucose or a galactose to Haemophilus influenzae lipopolysaccharide. AB - Nontypeable Haemophilus influenzae is a commensal that frequently causes otitis media and respiratory tract infections. The lex2 locus encodes a glycosyltransferase that is phase variably expressed and contributes to the significant intrastrain heterogeneity of lipopolysaccharide (LPS) composition in H. influenzae. In serotype b strains, Lex2B adds the second beta-glucose in the oligosaccharide extension from the proximal heptose of the triheptose inner core backbone; this extension includes a digalactoside that plays a role in resistance of the bacteria to the killing effect of serum. As part of our studies of the structure and genetics of LPS in nontypeable H. influenzae, we show here that there are allelic polymorphisms in the lex2B sequence that correlate with addition of either a glucose or a galactose to the same position in the LPS molecule across strains. Through exchange of lex2 alleles between strains we show that alteration of a single amino acid at position 157 in Lex2B appears to be sufficient to direct the alternative glucosyl- or galactosyltransferase activities. Allelic exchange strains express LPS with altered structure and biological properties compared to the wild-type LPS. Thus, Lex2B contributes to both inter- and intrastrain LPS heterogeneity through its polymorphic sequences and phase-variable expression. PMID- 19289513 TI - Resistance of Haemophilus influenzae to reactive nitrogen donors and gamma interferon-stimulated macrophages requires the formate-dependent nitrite reductase regulator-activated ytfE gene. AB - Haemophilus influenzae efficiently colonizes and persists at the human nasopharyngeal mucosa, causing disease when it spreads to other sites. Nitric oxide (NO) represents a major antimicrobial defense deployed by host cells in locations colonized by H. influenzae during pathogenesis that are likely to vary in oxygen levels. Formate-dependent nitrite reductase regulator (FNR) is an oxygen-sensitive regulator in several bacterial pathogens. We report that fnr of H. influenzae is required for anaerobic defense against exposure to NO donors and to resist NO-dependent effects of gamma interferon (IFN-gamma)-activated murine bone marrow-derived macrophages. To understand the mechanism of resistance, we investigated the role of FNR-regulated genes in defense against NO sources. Expression analysis revealed FNR-dependent activation of nrfA, dmsA, napA, and ytfE. Nonpolar deletion mutants of nrfA and ytfE exhibited sensitivity to NO donors, and the ytfE gene was more critical for survival. Compared to the wild type strain, the ytfE mutant exhibited decreased survival when exposed to macrophages, a defect that was more pronounced after prior stimulation of macrophages with IFN-gamma or lipopolysaccharide. Complementation restored survival of the mutant to the level in the parental strain. Increased sensitivity of the ytfE mutant relative to that of the parent was abrogated by treatment of macrophages with a NO synthase inhibitor, implicating YtfE in resistance to a NO dependent pathway. These results identify a requirement for FNR in positive control of ytfE and indicate a critical role for ytfE in resistance of H. influenzae to reactive nitrogen species and the antibacterial effects of macrophages. PMID- 19289514 TI - A comprehensive survey of single nucleotide polymorphisms (SNPs) across Mycobacterium bovis strains and M. bovis BCG vaccine strains refines the genealogy and defines a minimal set of SNPs that separate virulent M. bovis strains and M. bovis BCG strains. AB - To further unravel the mechanisms responsible for attenuation of the tuberculosis vaccine Mycobacterium bovis BCG, comparative genomics was used to identify single nucleotide polymorphisms (SNPs) that differed between sequenced strains of Mycobacterium bovis and M. bovis BCG. SNPs were assayed in M. bovis isolates from France and the United Kingdom and from different BCG vaccines in order to identify those that arose during the attenuation process which gave rise to BCG. Informative data sets were obtained for 658 SNPs from 21 virulent M. bovis strains and 13 BCG strains; these SNPs showed phylogenetic clustering that was consistent with the geographical origin of the strains and previous schemes for BCG genealogies. The data revealed a closer relationship between BCG Tice and BCG Pasteur than was previously appreciated, while we were able to position BCG Beijing within a grouping of BCG Denmark-derived strains. Only 186 SNPs were identified between virulent M. bovis strains and all BCG strains, with 115 nonsynonymous SNPs affecting important functions such as global regulators, transcriptional factors, and central metabolism, which might impact on virulence. We therefore refine previous genealogies of BCG vaccines and define a minimal set of SNPs between virulent M. bovis strains and the attenuated BCG strain that will underpin future functional analyses. PMID- 19289515 TI - Role of HrpA in biofilm formation of Neisseria meningitidis and regulation of the hrpBAS transcripts. AB - Two-partner secretion systems of gram-negative organisms are utilized in adherence, invasion, and biofilm formation. The HrpAB proteins of Neisseria meningitidis are members of a two-partner secretion system, and HrpA is established as being important to adherence and intracellular escape. This study set out to determine the expression pattern of members of the hrpBAS putative operon and to find a functional role for the HrpA protein. The upregulation of these genes was found in situations of anaerobiosis and cell contact. These observations prompted the study of the function of HrpA in biofilms on human bronchial epithelial cells. HrpA mutants in encapsulated and unencapsulated NMB strains demonstrated biofilm growth equivalent to that of the wild-type strain at 6 h but a decreased ability to form biofilms at 48 h. Biofilms formed by hrpA mutants for 48 h on collagen-coated coverslips demonstrated significant reductions compared to those of wild-type strains. Taken together, these observations imply a role for HrpA in the biofilm structure. Further analysis demonstrated the presence of HrpA on the surface of the bacterium. PMID- 19289516 TI - Genetically modified L3,7 and L2 lipooligosaccharides from Neisseria meningitidis serogroup B confer a broad cross-bactericidal response. AB - Currently available Neisseria meningitidis serogroup B (MenB) vaccines are based on outer membrane vesicles (OMVs) that are obtained from wild-type strains. They are purified with the aim of decreasing the lipooligosaccharide (LOS) content and hence reduce the reactogenicity of the vaccine even though LOS is a potential protective antigen. In <2-year-old children, these MenB vaccines confer protection only against strains expressing homologous PorA, a major and variable outer membrane protein. Our objective was to develop a safe LOS-based vaccine against MenB. To this end, we used modified porA knockout strains expressing genetically detoxified (msbB gene-deleted) L2 and L3,7 LOSs, allowing the production of LOS-enriched OMVs. The vaccine-induced antibodies were found to be bactericidal against nearly all invasive strains, irrespective of capsular serogroup. In addition, we have also demonstrated that LOS lacking the terminal galactose (with a lgtB mutation; truncated L3 LOS), but not LOS produced without the galE gene, induced a bactericidal antibody response in mice similar to that seen for LOS containing the full lacto-N-neotetraose (L3,7 LOS). In conclusion, a bivalent detoxified LOS OMV-based vaccine demonstrated the potential to afford a broad cross-protection against meningococcal disease. PMID- 19289517 TI - Regulation of mprF in daptomycin-nonsusceptible Staphylococcus aureus strains. AB - We used a well-characterized isogenic set of clinical bloodstream Staphylococcus aureus strains to study (i) regulation of mprF-mediated phosphatidylglycerol lysinylation in the contexts of in vitro daptomycin (DAP) nonsuceptibility and (ii) the role of mprF mutation in endovascular virulence. We observed a correlation between increased expression of a mutant mprF gene and reduced in vitro DAP susceptibility. There were no detectable fitness differences between strains in experimental infective endocarditis. PMID- 19289518 TI - The octadecyloxyethyl ester of (S)-9-[3-hydroxy-2-(phosphonomethoxy) propyl]adenine is a potent and selective inhibitor of hepatitis C virus replication in genotype 1A, 1B, and 2A replicons. AB - The octadecyloxyethyl (ODE) and hexadecyloxypropyl (HDP) esters of (S)-9-[3 hydroxy-2-(phosphonomethoxy)propyl]adenine (HPMPA) are potent inhibitors of orthopoxvirus, herpesvirus, human immunodeficiency virus type 1, and hepatitis B virus replication in vitro. HDP and ODE esters of (S)-HPMPA and (R)-HPMPA were evaluated for their activity in hepatitis C virus (HCV) replicon assays using luciferase (1B and 2A replicons) or RNA (1B) quantification. The ODE ester of (S) HPMPA [ODE-(S)-HPMPA] was the most active compound, with 50% effective concentrations (EC(50)s) in the 0.69 to 1.31 microM range. HDP and ODE esters of (R)-HPMPA were severalfold less active, while (S)-HPMPA and (R)-HPMPA were inactive. In genotype 1A and 1B replicons analyzed by HCV RNA analysis, ODE-(S) HPMPA was the most active compound, with EC(50)s of 1.8 and 2.1 microM, respectively. PMID- 19289519 TI - Prevention of death in bacterium-infected mice by a synthetic antimicrobial peptide, L5, through activation of host immunity. AB - In our previous study, we found that the antibacterial peptide KLKLLLLLKLK-NH(2) (L5) and its d-enantiomer (DL5) activate neutrophils to produce superoxide anions (O(2)(-)) and prevent death due to infection by methicillin-resistant Staphylococcus aureus, suggesting that these peptides may elicit in vivo antimicrobial activities through host inflammatory responses mediated by neutrophils. In this study, we investigated the mechanisms behind in vivo antimicrobial prophylaxis by the use of L5 for the treatment of bacterial infection introduced via intra-abdominal implantation. We found that the intraperitoneal treatment with L5 before bacterial infection markedly reduced rates of death due to infection. Treatments with L5 were highly effective in preventing death due to intraperitoneal inoculation of not only S. aureus Smith but also Enterococcus faecalis SR1004 and Escherichia coli EC14. The intra abdominal administration of L5 induced accumulation of neutrophils, increased levels of reactive oxygen species, and augmented antibacterial activity in the abdominal cavity. In addition, administration of L5 upregulated the expression of the Mig/CXCL9 chemokine gene in thioglycolate-elicited peritoneal macrophages. Our results suggested that the prevention of death by treatment of infected mice with L5 might occur primarily through the activation of a host immune response. PMID- 19289520 TI - Quasiexperimental study of the effects of antibiotic use, gastric acid suppressive agents, and infection control practices on the incidence of Clostridium difficile-associated diarrhea in hospitalized patients. AB - The objective of this study was to evaluate the effects of antimicrobial drug use, gastric acid-suppressive agent use, and infection control practices on the incidence of Clostridium difficile-associated diarrhea (CDAD) in a 426-bed general teaching hospital in Northern Ireland. The study was retrospective and ecological in design. A multivariate autoregressive integrated moving average (time-series analysis) model was built to relate CDAD incidence with antibiotic use, gastric acid-suppressive agent use, and infection control practices within the hospital over a 5-year period (February 2002 to March 2007). The findings of this study showed that temporal variation in CDAD incidence followed temporal variations in expanded-spectrum cephalosporin use (average delay = 2 months; variation of CDAD incidence = 0.01/100 bed-days), broad-spectrum cephalosporin use (average delay = 2 months; variation of CDAD incidence = 0.02/100 bed-days), fluoroquinolone use (average delay = 3 months; variation of CDAD incidence = 0.004/100 bed-days), amoxicillin-clavulanic acid use (average delay = 1 month; variation of CDAD incidence = 0.002/100 bed-days), and macrolide use (average delay = 5 months; variation of CDAD incidence = 0.002/100 bed-days). Temporal relationships were also observed between CDAD incidence and use of histamine-2 receptor antagonists (H2RAs; average delay = 1 month; variation of CDAD incidence = 0.001/100 bed-days). The model explained 78% of the variance in the monthly incidence of CDAD. The findings of this study highlight a temporal relationship between certain classes of antibiotics, H2RAs, and CDAD incidence. The results of this research can help hospitals to set priorities for restricting the use of specific antibiotic classes, based on the size-effect of each class and the delay necessary to observe an effect. PMID- 19289521 TI - Differing effects of combination chemotherapy with meropenem and tobramycin on cell kill and suppression of resistance of wild-type Pseudomonas aeruginosa PAO1 and its isogenic MexAB efflux pump-overexpressed mutant. AB - The drug interaction terminology (synergy, additivity, antagonism) relates to bacterial kill. The suppression of resistance requires greater drug exposure. We examined the combination of meropenem and tobramycin for kill and resistance suppression (wild-type Pseudomonas aeruginosa PAO1 and its isogenic MexAB overexpressed mutant). The drug interaction was additive. The introduction of MexAB overexpression significantly altered the 50% inhibitory concentration of meropenem but not that of tobramycin, resulting in the recovery of a marked increase in colony numbers from drug-containing plates. For the wild type, more tobramycin-resistant isolates than meropenem-resistant isolates were present, and the tobramycin-resistant isolates were harder to suppress. MexAB overexpression unexpectedly caused a significant increase in the number of tobramycin-resistant mutants, as indexed to the area under the curve of slices through the inverted U resistance mountain. The differences were significant, except in the absence of meropenem. We hypothesize that the pump resulted in the presence of less meropenem for organism inhibition, allowing more rounds of replication and also affecting the numbers of tobramycin-resistant mutants. When resistance suppression is explored by combination chemotherapy, it is important to examine the impacts of differing resistance mechanisms for both agents. PMID- 19289522 TI - Antiviral activity of MK-4965, a novel nonnucleoside reverse transcriptase inhibitor. AB - Nonnucleoside reverse transcriptase inhibitors (NNRTIs) are the mainstays of therapy for the treatment of human immunodeficiency virus type 1 (HIV-1) infections. However, the effectiveness of NNRTIs can be hampered by the development of resistance mutations which confer cross-resistance to drugs in the same class. Extensive efforts have been made to identify new NNRTIs that can suppress the replication of the prevalent NNRTI-resistant viruses. MK-4965 is a novel NNRTI that possesses both diaryl ether and indazole moieties. The compound displays potency at subnanomolar concentrations against wild-type (WT), K103N, and Y181C reverse transcriptase (RT) in biochemical assays. MK-4965 is also highly potent against the WT virus and two most prevalent NNRTI-resistant viruses (viruses that harbor the K103N or the Y181C mutation), against which it had 95% effective concentrations (EC(95)s) of <30 nM in the presence of 10% fetal bovine serum. The antiviral EC(95) of MK-4965 was reduced approximately four- to sixfold when it was tested in 50% human serum. Moreover, MK-4965 was evaluated with a panel of 15 viruses with NNRTI resistance-associated mutations and showed a superior mutant profile to that of efavirenz but not to that of etravirine. MK 4965 was similarly effective against various HIV-1 subtypes and viruses containing nucleoside reverse transcriptase inhibitor or protease inhibitor resistance-conferring mutations. A two-drug combination study showed that the antiviral activity of MK-4965 was nonantagonistic with each of the 18 FDA licensed drugs tested vice versa in the present study. Taken together, these in vitro data show that MK-4965 possesses the desired properties for further development as a new NNRTI for the treatment of HIV-1 infection. PMID- 19289523 TI - Inhaled voriconazole for prevention of invasive pulmonary aspergillosis. AB - Targeted airway delivery of antifungals as prophylaxis against invasive aspergillosis may lead to high lung drug concentrations while avoiding toxicities associated with systemically administered agents. We evaluated the effectiveness of aerosolizing the intravenous formulation of voriconazole as prophylaxis against invasive pulmonary aspergillosis caused by Aspergillus fumigatus in an established murine model. Inhaled voriconazole significantly improved survival and limited the extent of invasive disease, as assessed by histopathology, compared to control and amphotericin B treatments. PMID- 19289524 TI - Therapeutic evaluation of polyamine analogue drug candidates against Enterocytozoon bieneusi in a SCID mouse model. AB - Enterocytozoon bieneusi is the most common cause of chronic diarrhea in individuals with human immunodeficiency virus infection or AIDS, and there is no effective therapy. The inhibitory activities of polyamine analogues (PG-11157, PG 11158, and PG-11302) against E. bieneusi infection were evaluated in SCID mice preconditioned with anti-gamma interferon monoclonal antibody intraperitoneally (i.p.). Mice were challenged orally with 10(4) E. bieneusi spores, and groups of mice were treated orally or i.p. 14 days later for 7 days. The inhibitory activities of the drugs against infection were determined by enumerating the E. bieneusi spores in feces three times a week by an immunofluorescence assay. Immunohistochemistry staining confirmed the infection within enterocytes. Oral administration of the analogues PG-11157 (at 150 or 75 mg/kg of body weight/day) and PG-11302 (at 250 mg/kg/day) had significant inhibitory activity (96.2 to 99.6%) that was slightly better than that of fumagillin (1 mg/kg/day; 93.7%). The inhibitory activity with i.p. injection was significant only with PG-11302 at 20 mg/kg/day. While the treatments considerably reduced the levels of spore excretion, neither polyamine analogues nor fumagillin was able to completely eliminate E. bieneusi, as excretion reappeared within 7 days after the end of treatment. Drug toxicity was apparent during treatment, but it disappeared at the end of treatment. These results warrant further examination of the analogues PG 11157 and PG-11302. PMID- 19289525 TI - Activities of ceftobiprole and other cephalosporins against extracellular and intracellular (THP-1 macrophages and keratinocytes) forms of methicillin susceptible and methicillin-resistant Staphylococcus aureus. AB - Staphylococcus aureus is an opportunistic intracellular organism. Although they poorly accumulate in eukaryotic cells, beta-lactams show activity against intracellular methicillin (methicillin)-susceptible S. aureus (MSSA) if the exposure times and the drug concentrations are sufficient. Intraphagocytic methicillin-resistant S. aureus (MRSA) strains are susceptible to penicillins and carbapenems because the acidic pH favors the acylation of PBP 2a by these beta lactams through pH-induced conformational changes. The intracellular activity (THP-1 macrophages and keratinocytes) of ceftobiprole, which shows almost similar in vitro activities against MRSA and MSSA in broth, was examined against a panel of hospital-acquired and community-acquired MRSA strains (MICs, 0.5 to 2.0 mg/liter at pH 7.4 and 0.25 to 1.0 mg/liter at pH 5.5) and was compared with its activity against MSSA isolates. The key pharmacological descriptors {relative maximal efficacy (E(max)), relative potency (the concentration causing a reduction of the inoculum halfway between E(0) and E(max) [EC(50)]), and static concentration (C(s))} were measured. All strains showed sigmoidal dose-responses, with E(max) being about a 1 log(10) CFU decrease from the postphagocytosis inoculum, and EC(50) and C(s) being 0.2 to 0.3x and 0.6 to 0.9x the MIC, respectively. Ceftobiprole effectively competed with Bocillin FL (a fluorescent derivative of penicillin V) for binding to PBP 2a at both pH 5.5 and pH 7.4. In contrast, cephalexin, cefuroxime, cefoxitin, or ceftriaxone (i) were less potent in PBP 2a competitive binding assays, (ii) showed only partial restoration of the activity against MRSA in broth at acidic pH, and (iii) were collectively less effective against MRSA in THP-1 macrophages and were ineffective in keratinocytes. The improved activity of ceftobiprole toward intracellular MRSA compared with the activities of conventional cephalosporins can be explained, at least in part, by its greater ability to bind to PBP 2a not only at neutral but also at acidic pH. PMID- 19289526 TI - Prevalence of aac(6')-Ib-cr encoding a ciprofloxacin-modifying enzyme among Enterobacteriaceae blood isolates in Korea. AB - The aac(6')-Ib gene was detected in 86 of 555 (15.5%) Enterobacteriaceae isolates. Among these 86 aac(6')-Ib-positive isolates, 19 (22.0%) were positive for aac(6')-Ib-cr: 4 of 31 (12.9%) Enterobacter spp., 7 of 13 (53.8%) Escherichia coli isolates, and 8 of 42 (19.0%) Klebsiella pneumoniae isolates. There was a strong association between aac(6')-Ib-cr and OXA-1 and CTX-M-1 group beta lactamase genes. One aac(6')-Ib-positive K. pneumoniae isolate carried both aac(6')-Ib-cr and qnrS. PMID- 19289527 TI - Impact of sarA on antibiotic susceptibility of Staphylococcus aureus in a catheter-associated in vitro model of biofilm formation. AB - Mutation of the staphylococcal accessory regulator (sarA) in Staphylococcus aureus limits but does not abolish the capacity of the organism to form a biofilm. As a first step toward determining whether this limitation is therapeutically relevant, we carried out in vitro studies comparing the relative susceptibility of an S. aureus clinical isolate (UAMS-1) and its isogenic sarA mutant (UAMS-929) in the specific context of a catheter-associated biofilm. The antibiotics tested were daptomycin, linezolid, and vancomycin, all of which were evaluated by using concentrations based on the MIC defined as the breakpoint for a susceptible strain of S. aureus (< or = 1.0, < or = 2.0, and < or = 4.0 microg/ml for daptomycin, vancomycin, and linezolid, respectively). Mutation of sarA had no significant impact on the MIC of UAMS-1 for any of the targeted antibiotics, as defined by Etest antimicrobial susceptibility testing. However, mutation of sarA did result in a significant increase in antimicrobial susceptibility to all targeted antibiotics when they were tested in the specific context of a biofilm. Additionally, whether susceptibility was assessed by using UAMS-1 or its sarA mutant, daptomycin was found to be more effective against established S. aureus biofilms than either linezolid or vancomycin. PMID- 19289528 TI - Potency and bactericidal activity of iclaprim against recent clinical gram positive isolates. AB - The in vitro activity of iclaprim, a novel diaminopyrimidine derivative, was evaluated against 5,937 recent gram-positive clinical isolates collected in the United States and Europe. Iclaprim demonstrated potent activity against Staphylococcus aureus (including methicillin-resistant S. aureus [MRSA]), beta hemolytic Streptococcus spp., and Enterococcus faecalis strains tested. In addition, iclaprim exhibited bactericidal activity against all S. aureus strains tested, including MRSA. PMID- 19289529 TI - Genetic determinants of resistance to fusidic acid among clinical bacteremia isolates of Staphylococcus aureus. AB - Resistance to fusidic acid in Staphylococcus aureus is caused by mutation of the elongation factor G (EF-G) drug target (FusA class) or by expression of a protein that protects the drug target (FusB and FusC classes). Recently, two novel genetic classes of small-colony variants (SCVs) were identified among fusidic acid-resistant mutants selected in vitro (FusA-SCV and FusE classes). We analyzed a phylogenetically diverse collection of fusidic acid-resistant bacteremia isolates to determine which resistance classes were prevalent and whether these were associated with particular phylogenetic lineages. Each isolate was shown by DNA sequencing and plasmid curing to carry only one determinant of fusidic acid resistance, with approximately equal frequencies of the FusA, FusB, and FusC genetic classes. The FusA class (mutations in fusA) were distributed among different phylogenetic types. Two distinct variants of the FusC class (chromosomal fusC gene) were identified, and FusC was also distributed among different phylogenetic types. In contrast, the FusB class (carrying fusB on a plasmid) was found in closely related types. No FusE-class mutants (carrying mutations in rplF) were found. However, one FusA-class isolate had multiple mutations in the fusA gene, including one altering a codon associated with the FusA-SCV class. SCVs are frequently unstable and may undergo compensatory evolution to a normal growth phenotype after their initial occurrence. Accordingly, this normal-growth isolate might have evolved from a fusidic acid resistant SCV. We conclude that at least three different resistance classes are prevalent among fusidic acid-resistant bacteremia isolates of S. aureus. PMID- 19289530 TI - Novel S-adenosylmethionine decarboxylase inhibitors for the treatment of human African trypanosomiasis. AB - Trypanosomiasis remains a significant disease across the sub-Saharan African continent, with 50,000 to 70,000 individuals infected. The utility of current therapies is limited by issues of toxicity and the need to administer compounds intravenously. We have begun a program to pursue lead optimization around MDL 73811, an irreversible inhibitor of S-adenosylmethionine decarboxylase (AdoMetDC). This compound is potent but in previous studies cleared rapidly from the blood of rats (T. L. Byers, T. L. Bush, P. P. McCann, and A. J. Bitonti, Biochem. J. 274:527-533). One of the analogs synthesized (Genz-644131) was shown to be highly active against Trypanosoma brucei rhodesiense in vitro (50% inhibitory concentration, 400 pg/ml). Enzyme kinetic studies showed Genz-644131 to be approximately fivefold more potent than MDL 73811 against the T. brucei brucei AdoMetDC-prozyme complex. This compound was stable in vitro in rat and human liver microsomal and hepatocyte assays, was stable in rat whole-blood assays, did not significantly inhibit human cytochrome P450 enzymes, had no measurable efflux in CaCo-2 cells, and was only 41% bound by serum proteins. Pharmacokinetic studies of mice following intraperitoneal dosing showed that the half-life of Genz-644131 was threefold greater than that of MDL 73811 (7.4 h versus 2.5 h). Furthermore, brain penetration of Genz-644131 was 4.3-fold higher than that of MDL 73811. Finally, in vivo efficacy studies of T. b. brucei strain STIB 795-infected mice showed that Genz-644131 significantly extended survival (from 6.75 days for controls to >30 days for treated animals) and cured animals infected with T. b. brucei strain LAB 110 EATRO. Taken together, the data strengthen validation of AdoMetDC as an important parasite target, and these studies have shown that analogs of MDL 73811 can be synthesized with improved potency and brain penetration. PMID- 19289531 TI - In Vitro activity of terbinafine combined with caspofungin and azoles against Pythium insidiosum. AB - In this text we evaluated the in vitro antifungal activities of terbinafine combined with caspofungin, miconazole, ketoconazole, and fluconazole against 17 Pythium insidiosum strains by using the microdilution checkerboard method. Synergistic interactions were observed with terbinafine combined with caspofungin (41.2% of the strains), fluconazole (41.2%), ketoconazole (29.4%), and miconazole (11.8%). No antagonistic effects were observed. The combination of terbinafine plus caspofungin or terbinafine plus fluconazole may have significant therapeutic potential for treatment of pythiosis. PMID- 19289532 TI - HR11 and HR13 receptor-expressing neurons are housed together in pheromone responsive sensilla trichodea of male Heliothis virescens. AB - The highly specific recognition of female-released sex pheromones in insects by sensory neurons of the male antenna requires specific receptors. Recently, a small family of related candidate pheromone receptors has been identified for a few moth species. In this study, the candidate pheromone receptor HR11 from Heliothis virescens has been characterized. HR11 was found to be expressed in numerous cells located in short and long sensilla trichodea on the male antenna. The HR11 cells are stereotypically arranged in a paired pattern together with HR13 cells, which respond to the major component of the sex pheromone blend. Triple in situ hybridization approaches revealed that each pair of an HR11 cell and an HR13 cell was ensheathed by supporting cells, which express pheromone binding proteins, thus constituting a structural unit. The paired pattern of HR11/HR13 cells is reminiscent of the pattern described for BmOR-1- and BmOR-3 expressing cells in the antenna of Bombyx mori, which respond to bombykol and bombykal, respectively. These results suggest that the ligand for HR11 may be related to the HR13 ligand and furthermore imply that an arrangement of cells expressing related receptor types in the same sensillum may be a general principle in moth pheromone detection systems. PMID- 19289533 TI - Pheochromocytomas and extra-adrenal paragangliomas detected by screening in patients with SDHD-associated head-and-neck paragangliomas. AB - Patients with SDHD-associated head-and-neck paragangliomas (HNP) are at risk for developing pheochromocytomas for which screening has been advised. To assess clinical, biochemical, and radiological outcomes of screening in a large single center cohort of SDHD-positive patients with HNP and to address the necessity for repetitive follow-up, we evaluated 93 patients with SDHD-associated HNP (p.Asp92Tyr, p.Leu139Pro). Screening consisted of measurement of 24 h urinary excretion of catecholamines and/or their metabolites in duplicate, which was repeated with intervals of 2 years if initial biochemical screening was negative. In patients, in whom urinary excretion was above the reference limit, imaging studies with (123)I-MIBG (metaiodobenzylguanidine) scintigraphy and magnetic resonance imaging (MRI) and/or computed tomography (CT) were performed. Pheochromocytomas and extra-adrenal paragangliomas were treated surgically after appropriate blockade. Median follow-up was 4.5 years (range 0.5-19.5 years). Twenty-eight out of the 93 patients were included in our study and underwent additional imaging for pheochromocytomas/extra-adrenal paragangliomas. In 11 out of the 28 patients intra-adrenal pheochromocytomas were found. Extra-adrenal paragangliomas were discovered in eight patients. These tumors were detected during initial screening in 63% of cases, whereas 37% were detected after repeated biochemical screening. One patient was diagnosed with a biochemically silent pheochromocytoma. The high prevalence of pheochromocytomas/extra-adrenal paragangliomas in patients with SDHD-associated HNP warrants regular screening for tumors in these patients. Paragangliomas that do not secrete catecholamines might be more prevalent than previously reported. Future studies will have to establish whether routine imaging studies should be included in the screening of SDHD mutation carriers, irrespective of biochemical screening. PMID- 19289534 TI - Merits and pitfalls of mifepristone in Cushing's syndrome. AB - OBJECTIVE: Mifepristone is the only available glucocorticoid receptor antagonist. Only few adult patients with hypercortisolism were treated to date by this drug. Our objective was to determine effectiveness and tolerability of mifepristone in Cushing's syndrome (CS). DESIGN: Retrospective study of patients treated in seven European centers. METHODS: Twenty patients with malignant (n=15, 12 with adrenocortical carcinoma, three with ectopic ACTH secretion) or benign (n=5, four with Cushing's disease, one with bilateral adrenal hyperplasia) CS were treated with mifepristone. Mifepristone was initiated with a median starting dose of 400 mg/day (200-1000). Median treatment duration was 2 months (0.25-21) for malignant CS, and 6 months (0.5-24) for benign CS. Clinical (signs of hypercortisolism, blood pressure, signs of adrenal insufficiency), and biochemical parameters (serum potassium and glucose) were evaluated. RESULTS: Treatment was stopped in one patient after 1 week due to severe uncontrolled hypokalemia. Improvement of clinical signs was observed in 11/15 patients with malignant CS (73%), and 4/5 patients with benign CS (80%). Psychiatric symptoms improved in 4/5 patients within the first week. Blood glucose levels improved in 4/7 patients. Signs of adrenal insufficiency were observed in 3/20 patients. Moderate to severe hypokalemia was observed in 11/20 patients and increased blood pressure levels in 3/20 patients. CONCLUSION: Mifepristone is a rapidly effective treatment of hypercortisolism, but requires close monitoring of potentially severe hypokalemia, hypertension, and clinical signs of adrenal insufficiency. Mifepristone provides a valuable treatment option in patients with severe CS when surgery is unsuccessful or impossible. PMID- 19289535 TI - Vitamin D in relation to metabolic risk factors, insulin sensitivity and adiponectin in a young Middle-Eastern population. AB - OBJECTIVES: Several studies suggest a link between circulating 25-hydroxyvitamin D (25(OH)D) and metabolic risk factors. However, this relation has been mainly studied in elderly and/or obese subjects. In addition, the relation between 25(OH)D and adiponectin is unclear. The purpose of this study is to look at these relations in non-obese young individuals. DESIGN: We investigated the relation between serum 25(OH)D and adiposity, blood pressure, glucose metabolism, lipid profile, and adiponectin in 381 randomly selected university students (201 males and 180 females, mean age 23.9+/-3.9). RESULTS: In the overall population, 25(OH)D is significantly inversely correlated with body mass index (BMI), systolic blood pressure (SBP), waist circumference (WC), fasting plasma glucose (FPG), insulin levels, and homeostasis model assessment of insulin resistance (HOMA index) and positively correlated with adiponectin and high density lipoprotein-cholesterol (P<0.01 for all variables). In males, these correlations are still significant for BMI, SBP, WC, and adiponectin (P=0.02, P=0.01, P=0.04 and P=0.01 respectively); also, 25(OH)D is inversely correlated with low density lipoprotein (LDL)-cholesterol (P=0.007). In females, 25(OH)D is only inversely correlated with FPG and HOMA index (P<0.001 and P=0.03 respectively). In multivariate regression analysis models, after adjustment for sex and BMI, 25(OH)D is an independent predictor of FPG and SBP (P=0.032 and P=0.05 respectively) in the overall population, while in males 25(OH)D is a predictor of LDL-cholesterol and SBP independently of BMI (P=0.007 and P=0.035 respectively). CONCLUSION: In non-obese young subjects, we observe new relationships between 25(OH)D and several metabolic risk factors and adiponectin. Further research is needed to elucidate the gender differences and to look at the relation between 25(OH)D and adiponectin. PMID- 19289536 TI - Obestatin/ghrelin cells in normal mucosa and endocrine tumours of the stomach. AB - OBJECTIVE: Obestatin and ghrelin are derived from the same gene and co-expressed in the same endocrine cells. Vesicular monoamine transporter-2 (VMAT-2), a marker for enterochromaffin-like (ECL) cells, is considered to be expressed in ghrelin cells. The aim was to establish if the two peptides and the transporter are co expressed, both in normal gastric mucosa and in gastric endocrine tumours. DESIGN: An immunohistochemical study was performed on gastric biopsy material and on surgical specimens from 63 patients with gastric endocrine tumours and from individuals with normal gastric mucosa. Cells displaying obestatin immunoreactivity were examined regarding co-localization with ghrelin and VMAT-2. Both single- and double-immunostaining techniques were applied. Obestatin concentration in blood was measured in a subgroup of these patients. The results were correlated to various clinico-pathological parameters. RESULTS: In the normal mucosa, obestatin/ghrelin-immunoreactive cells rarely co-expressed VMAT-2. In most tumour tissue specimens, only a fraction of neoplastic cells displayed immunoreactivity to obestatin, and these cells always co-expressed ghrelin. Neoplastic obestatin-/ghrelin-IR cells invariably expressed VMAT-2, except for two ghrelinomas. The obestatin concentrations in blood were consistently low and did not correlate to clinico-pathological data. CONCLUSIONS: Obestatin and ghrelin immunoreactivity always occurred in the same endocrine cells in the gastric mucosa but these cells only occasionally co-expressed VMAT-2, opposite to the findings in tumours. These results indicate that endocrine cells expressing obestatin and ghrelin mainly differ from VMAT-2 expressing cells (ECL-cells) and can develop into pure ghrelinomas. Plasma concentrations of obestatin did not correlate to cellular expression. PMID- 19289537 TI - The prevalence and characteristic features of cyclicity and variability in Cushing's disease. AB - OBJECTIVE: Cyclical Cushing's syndrome may render the diagnosis and management of Cushing's disease difficult. The aim of the present study was to investigate the prevalence of cyclicity and variability in patients with Cushing's disease, and to identify putative distinctive features. DESIGN: Retrospective case-note study. METHODS: We analysed the case records of 201 patients with Cushing's disease in a retrospective case-note study. Cyclicity was considered as the presence of at least one cycle, defined as a clinical and/or biochemical hypercortisolaemic peak followed by clinical and biochemical remission, followed by a new clinical and/or biochemical hypercortisolaemic peak. The fluctuations of mean serum cortisol levels, as assessed by a 5-point cortisol day curve, defined the variability. RESULTS: Thirty (14.9%; 26 females) patients had evidence of cyclicity/variability. 'Cycling' patients were older but no difference in sex or paediatric distribution was revealed between 'cycling' and 'non-cycling' patients. The median number of cycles was two for each patient, and 4 years was the median intercyclic period. A trend to lower cure rate post-neurosurgery and lower adenoma identification was observed in 'cycling' compared with 'non cycling' patients. In multivariate analysis, older patients, longer follow-up, female sex and no histological identification of the adenoma were associated with an increased risk of cyclic disease. CONCLUSIONS: This large population study reveals that cyclicity/variability is not an infrequent phenomenon in patients with Cushing's disease, with a minimum prevalence of 15%. Physicians should be alert since it can lead to frequent problems in diagnosis and management, and no specific features can be used as markers. PMID- 19289538 TI - Correlation between morphology and function of the upper lip: a longitudinal evaluation. AB - In order to evaluate the relationship between the morphology of the upper lip and muscle activity in a sample of 38 subjects (17 males and 21 females) with Angle Class II division 1 malocclusions, cephalometric and electromyographic analyses were conducted. The sample was subdivided into either predominantly nose or mouth breathers. The individuals were evaluated at two different periods, with a 2 year interval. At the first observation, the subjects were 11 years to 14 years 11 months of age and at the second observation, 13 years 4 months to 16 years 6 months of age. Height and thickness of the upper lip were measured on lateral cephalograms with the aid of a digital pachymeter. For each individual, electromyographic records were obtained of the orbicularis oris superior muscle at rest and in a series of 12 movements. The electromyographic data were normalized as a function of amplitude, for achievement of the percentage value of each movement. Pearson and Spearman correlation tests were applied. The results showed some correlation between morphology and muscle function (at a confidence level of 95 per cent). However, as the values of the correlation coefficient (r) were too low to establish associations between variables, it was concluded that the dimensions of the upper lip are not correlated with muscle activity. PMID- 19289539 TI - Miniscrews for upper incisor intrusion. AB - The aim of this study was to investigate if true incisor intrusion can be achieved using miniscrews. Eleven patients (three males and eight females; mean age: 19.8 +/- 4.8 years) with normal vertical dimension showing a pre-treatment deep bite of 5.9 +/- 0.9 mm and a 'gummy' smile were enrolled in the study. After levelling of the maxillary central and lateral incisors with a segmental arch, an intrusive force of 80 g using closed coil springs was applied from two miniscrews placed between the roots of the lateral and canine teeth. The amount of incisor intrusion was evaluated on lateral cephalometric headfilms taken at the end of levelling (T1) and at the end of intrusion (T2). Statistical analysis of the data was performed using a paired t and Wilcoxon signed rank tests. A significance level of P < 0.05 was predetermined. The mean upper incisor intrusion was 1.92 mm and the mean overbite decrease 2.25 +/- 1.73 mm in 4.55 months. Upper incisor angulation resulted in a 1.81 +/- 3.84 degree change in U1-PP angle and a 1.22 +/ 3.64 degree change in U1-NA angle. However, these were not statistically significant (P > 0.05). True intrusion can be achieved by application of intrusive forces close to the centre of resistance using miniscrews. However, studies with a larger number of subjects and long-term follow-up are necessary. PMID- 19289540 TI - Palatally impacted canines and the modified index of orthodontic treatment need. AB - The aim of this study was to assess the severity of any underlying malocclusion in subjects presenting for treatment of a palatally impacted canine (PIC) using a modification of the Dental Health Component (DHC) of the Index of Treatment Need (MIOTN), which does not factor in the impacted canine. The pre-treatment study models of 54 subjects who had previously undergone surgical exposure of a PIC, followed by fixed appliance orthodontic alignment, were scored independently by two examiners on two occasions using the MIOTN system. Unweighted kappa statistics revealed good intraoperator agreement for the two examiners and a moderate level of interexaminer agreement. Forty-six and 41 per cent of the sample still scored either an MIOTN grade 4 or 5 (i.e. a great or very great need of orthodontic treatment). However, 20 and 25 per cent of the subjects were graded with a MIOTN score of 1 or 2, indicating little or no need for treatment when the PIC was not taken into consideration. This finding emphasizes the importance of early diagnosis of an impacted canine and the need to institute interceptive measures where necessary, as up to 25 per cent of patients might otherwise require no other orthodontic treatment. PMID- 19289541 TI - Alterations in knee joint laxity during the menstrual cycle in healthy women leads to increases in joint loads during selected athletic movements. AB - BACKGROUND: It has been speculated that the hormonal cycle may be correlated with higher incidence of ACL injury in female athletes, but results have been very contradictory. HYPOTHESIS: Knee joint loads are influenced by knee joint laxity (KJL) during the menstrual cycle. STUDY DESIGN: Controlled laboratory study. METHODS: Serum samples and KJL were assessed at the follicular, ovulation, and luteal phases in 26 women. Knee joint mechanics (angle, moment, and impulse) were measured and compared at the same intervals. Each of the 26 subjects had a value for knee laxity at each of the 3 phases of their cycle, and these were ordered and designated low, medium, and high for that subject. Knee joint mechanics were then compared between low, medium, and high laxity. RESULTS: No significant differences in knee joint mechanics were found across the menstrual cycle (no phase effect). However, an increase in KJL was associated with higher knee joint loads during movement (laxity effect). A 1.3-mm increase in KJL resulted in an increase of approximately 30% in adduction impulse in a cutting maneuver, an increase of approximately 20% in knee adduction moment, and a 20% to 45% increase in external rotation loads during a jumping and stopping task (P < .05). CONCLUSION: Changes in KJL during the menstrual cycle do change knee joint loading during movements. Clinical Relevance Our findings will be beneficial for researchers in the development of more effective ACL injury prevention programs. PMID- 19289542 TI - Arthroscopic rotator cuff repair using the Opus knotless suture anchor fixation system. AB - BACKGROUND: The reported failure rate of arthroscopic rotator cuff repair varies widely. The influence of repair technique on failure rates and functional outcomes after arthroscopic cuff repair remains controversial. PURPOSE: To determine the functional outcome of arthroscopic knotless fixation using the Opus AutoCuff device for rotator cuff repair and to compare our results with those reported in the literature. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Fifty-six consecutive patients underwent arthroscopic rotator cuff repair using an Opus AutoCuff device (Arthrocare, Sunnydale, California) with greater than 2 years' follow-up. Subjective and objective clinical examinations were performed to include the University of California at Los Angeles (UCLA) shoulder score, the American Shoulder and Elbow Surgeons (ASES) rating scale, the visual analog scale (VAS), and the Tegner Activity Level scale. RESULTS: Forty eight patients were evaluated at a mean follow-up of 26 months (range, 24-35 months). The mean UCLA shoulder score was 33.1 of 35 (SD, 2.89) possible points, and the mean ASES rating scale was 94.2 of 100 (SD, 7.76) compared with a mean preoperative score of 65.7 (P < .001). Postoperative UCLA shoulder scores had 42 of 45 (93.3%) patients with good and excellent results. The mean preoperative ASES pain score was 1.3 (SD, 1.0), and the mean postoperative score was 4.4 (SD, 1.0) (P < .001). The Tegner Activity Level scores demonstrated restoration of function to preinjury status. There were 3 failures (6.3%), 2 by anchor failure (1 with specific trauma), and 1 by rotator cuff retear, all requiring revision surgery. CONCLUSION: Arthroscopic knotless suture fixation with the Opus AutoCuff device results in good to excellent results similar to those reported in the literature with conventional suture anchors. PMID- 19289543 TI - Whole grain and legume acceptability among youths with type 1 diabetes. AB - PURPOSE: This pilot study investigated the acceptability of whole grain and legume foods in youths with type 1 diabetes and determined demographic and behavioral factors associated with their acceptability. METHODS: Youths with type 1 diabetes (7.0 to 16.9 years) were recruited during a diabetes camp and completed self-report measures of food acceptability, eating behaviors, and demographics. An overall acceptability score for whole grains and for legumes was calculated as the sum of foods in each category rated as either "tried and liked" or "not tried and willing to try." Correlations of whole grain and legume acceptability with demographic characteristics were examined. RESULTS: One hundred twenty-eight youths participated (70% females, mean age 11.6 +/- 2.3 years). Whole grain foods with the highest acceptability were corn bread (85% tried/liked and 11% willing to try) and whole wheat bread (72% tried/liked and 3% willing to try). Total whole grain acceptability was negatively associated with days per week of fast food consumption (r = -0.21; P < .02) and living in an urban environment (r = -0.24; P < .01). Chili with beans (66% tried/liked and 8% willing to try) and baked beans (57% tried/liked and 19% willing to try) were the legume-containing foods with the highest acceptability. There were no significant associations between demographic factors and total legume acceptability. CONCLUSIONS: These findings demonstrate the variability in acceptability among whole grains and legumes in youths with type 1 diabetes and the importance of addressing acceptability when counseling youths or designing nutrition interventions to improve consumption of these foods. PMID- 19289544 TI - Sensitization of lamina I spinoparabrachial neurons parallels heat hyperalgesia in the chronic constriction injury model of neuropathic pain. AB - It has been proposed that spinal lamina I neurons with ascending axons that project to the midbrain play a crucial role in hyperalgesia. To test this hypothesis the quantitative properties of lamina I spinoparabrachial neurons in the chronic constriction injury (CCI) model of neuropathic pain were compared to those of unoperated and sham-operated controls. Behavioural testing showed that animals with a CCI exhibited heat hyperalgesia within 4 days of the injury, and this hyperalgesia persisted throughout the 14-day post-operative testing period. In the CCI, nociceptive lamina I spinoparabrachial neurons had heat thresholds that were significantly lower than controls (43.0 +/- 2.8 degrees C vs. 46.7 +/- 2.6 degrees C; P < 10(-4), ANOVA). Nociceptive lamina I spinoparabrachial neurons were also significantly more responsive to graded heat stimuli in the CCI, compared to controls (P < 0.02, 2-factor repeated-measures ANOVA), and increased after-discharges were also observed. Furthermore, the heat-evoked stimulus response functions of lamina I spinoparabrachial neurons in CCI animals co-varied significantly (P < 0.03, ANCOVA) with the amplitude of heat hyperalgesia determined behaviourally. Taken together these results are consistent with the hypothesis that lamina I spinoparabrachial neurons have an important mechanistic role in the pathophysiology of neuropathic pain. PMID- 19289545 TI - A role for nuclear receptors in mammalian hibernation. AB - Hibernation is one of the most dramatic examples of phenotypic plasticity in mammals. During periods of food shortage and/or reduced ambient temperatures hibernating mammals become heterothermic, allowing their body temperature to decrease while entering an energy-conserving torpid state. In order to survive the multi-month hibernation season many species engage in hyperphagy, dramatically increasing adipose stores prior to the onset of hibernation. Nuclear receptors are a superfamily of transcription factors many of which bind lipophilic molecules as ligands. They regulate a variety of processes including energy homeostasis, carbohydrate and lipid metabolism, inflammation and circadian rhythm. Given that lipids are integral in the hibernation phenotype they may play important regulatory roles through their interactions with nuclear receptors. Here we review current knowledge and suggest possible roles in mammalian hibernation for peroxisome proliferator-activated receptors (PPARs), farnesoid X receptors (FXRs), liver X receptors (LXRs), retinoid-related orphan receptors (RORs) and Rev-ERBs. PMID- 19289546 TI - Functional K(ATP) channels in the rat retinal microvasculature: topographical distribution, redox regulation, spermine modulation and diabetic alteration. AB - The essential task of the circulatory system is to match blood flow to local metabolic demand. However, much remains to be learned about this process. To better understand how local perfusion is regulated, we focused on the functional organization of the retinal microvasculature, which is particularly well adapted for the local control of perfusion. Here, we assessed the distribution and regulation of functional K(ATP) channels whose activation mediates the hyperpolarization induced by adenosine. Using microvascular complexes freshly isolated from the rat retina, we found a topographical heterogeneity in the distribution of functional K(ATP) channels; capillaries generate most of the K(ATP) current. The initiation of K(ATP)-induced responses in the capillaries supports the concept that the regulation of retinal perfusion is highly decentralized. Additional study revealed that microvascular K(ATP) channels are redox sensitive, with oxidants increasing their activity. Furthermore, the oxidant-mediated activation of these channels is driven by the polyamine spermine, whose catabolism produces oxidants. In addition, our observation that spermine-dependent oxidation occurs predominately in the capillaries accounts for why they generate most of the K(ATP) current detected in retinal microvascular complexes. Here, we also analysed retinal microvessels of streptozotocin-injected rats. We found that soon after the onset of diabetes, an increase in spermine dependent oxidation at proximal microvascular sites boosts their K(ATP) current and thereby virtually eliminates the topographical heterogeneity of functional K(ATP) channels. We conclude that spermine-dependent oxidation is a previously unrecognized mechanism by which this polyamine modulates ion channels; in addition to a physiological role, spermine-dependent oxidation may also contribute to microvascular dysfunction in the diabetic retina. PMID- 19289547 TI - Mapping of direction and muscle representation in the human primary motor cortex controlling thumb movements. AB - Larger body parts are somatotopically represented in the primary motor cortex (M1), while smaller body parts, such as the fingers, have partially overlapping representations. The principles that govern the overlapping organization of M1 remain unclear. We used transcranial magnetic stimulation (TMS) to examine the cortical encoding of thumb movements in M1 of healthy humans. We performed M1 mapping of the probability of inducing a thumb movement in a particular direction and used low intensity TMS to disturb a voluntary thumb movement in the same direction during a reaction time task. With both techniques we found spatially segregated representations of the direction of TMS-induced thumb movements, thumb flexion and extension being best separated. Furthermore, the cortical regions corresponding to activation of a thumb muscle differ, depending on whether the muscle functions as agonist or as antagonist for flexion or extension. In addition, we found in the reaction time experiment that the direction of a movement is processed in M1 before the muscles participating in it are activated. It thus appears that one of the organizing principles for the human corticospinal motor system is based on a spatially segregated representation of movement directions and that the representation of individual somatic structures, such as the hand muscles, overlap. PMID- 19289548 TI - Dynamic integration of subplate neurons into the cortical barrel field circuitry during postnatal development in the Golli-tau-eGFP (GTE) mouse. AB - In the Golli-tau-eGFP (GTE) transgenic mouse the reporter gene expression is largely confined to the layer of subplate neurons (SPn), providing an opportunity to study their intracortical and extracortical projections. In this study, we examined the thalamic afferents and layer IV neuron patterning in relation to the SPn neurites in the developing barrel cortex in GTE mouse at ages embryonic day 17 (E17) to postnatal day 14 (P14). Serotonin transporter immunohistochemistry or cytochrome oxydase histochemistry was used to reveal thalamic afferent patterning. Bizbenzimide staining identified the developing cytoarchitecture in coronal and tangential sections of GTE brains. Enhanced green fluorescent protein (GFP)-labelled neurites and thalamic afferents were both initially diffusely present in layer IV but by P4-P6 both assumed the characteristic periphery related pattern and became restricted to the barrel hollows. This pattern gradually changed and by P10 the GFP-labelled neurites largely accumulated at the layer IV-V boundary within the barrel septa whereas thalamic afferents remained in the hollows. To investigate whether this reorganisation is dependent on sensory input, the whiskers of row 'a' or 'c' were removed at P0 or P5 and the organisation of GFP-labelled neurites in the barrel cortex was examined at P6 or P10. In the contralateral region corresponding to row 'a' or 'c' the lack of hollow to septa rearrangement of the GFP-labelled neurites was observed after P0 row removal at P10 but not at P6. Our findings suggest a dynamic, sensory periphery-dependent integration of SPn neurites into the primary somatosensory cortex during the period of barrel formation. PMID- 19289549 TI - Cell volume and membrane stretch independently control K+ channel activity. AB - A number of potassium channels including members of the KCNQ family and the Ca(2+) activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch. To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (approximately 50 microm(2) macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current increases with increasing negative hydrostatic pressure (suction) applied to the pipette. Thus, at a pipette pressure of -5.0 +/- 0.1 mmHg the increase amounted to 381 +/- 146% (mean +/- S.E.M., n = 6, P < 0.025). In contrast, in oocytes expressing the strongly volume-sensitive KCNQ1 channel, the current was not affected by membrane stretch. The results indicate that (1) activation of BK channels by local membrane stretch is not mimicked by membrane stress induced by cell swelling, and (2) activation of KCNQ1 channels by cell volume increase is not mediated by local tension in the cell membrane. We conclude that stretch and volume sensitivity can be considered two independent regulatory mechanisms. PMID- 19289550 TI - The proprioceptive and agonist roles of gastrocnemius, soleus and tibialis anterior muscles in maintaining human upright posture. AB - Humans can stand using sensory information solely from the ankle muscles. Muscle length and tension in the calf muscles (gastrocnemius and soleus) are unlikely to signal postural sways on account of balance-related modulation in agonist activity. These facts pose two questions: (1) Which ankle muscles provide the proprioceptive information? (2) Which peripheral mechanism could modulate agonist activity? To address these issues, subjects were asked to stand normally on two force plates. Ultrasound and surface EMG were recorded from the calf and tibialis anterior (TA) muscles. For all nine subjects, changes in muscle length of TA were mainly (84 +/- 9% whole trial duration) orthodoxly correlated with bodily sway (centre of gravity, CoG), i.e. in accordance with passive ankle rotation. When orthodox, TA had the highest correlation with CoG (-0.66 +/- 0.07, deep compartment, P < 0.001). For five subjects, the superficial TA compartment showed counter-intuitive changes in muscle length with CoG, probably due to the flattening of the foot and proximal attachment geometry. Gastrocnemius and soleus were usually (duration 71 +/- 23 and 81 +/- 16%, respectively) active agonists (paradoxically correlated with CoG) but, for short periods of time, they could be orthodox and then presented a moderate correlation (0.38 +/- 0.16 and 0.28 +/- 0.09, respectively) with CoG. Considering the duration and extent to which muscle length is orthodox and correlated with CoG, TA may be a better source of proprioceptive information than the active agonists (soleus and gastrocnemius). Therefore, if a peripheral feedback mechanism modulates agonist activity then reciprocal inhibition acted by TA on the calf muscles is more likely to be effective than the autogenic pathway. PMID- 19289552 TI - 'Is it normal to feel these questions ...?': a content analysis of the health concerns of adolescent girls writing to a magazine. AB - BACKGROUND: There is a mismatch between presenting concerns of adolescents to GPs and behaviours that lead to adolescent morbidity and mortality. Better understanding of health concerns of this target group would enhance communication between health professionals and adolescent patients. OBJECTIVE: To explore and categorize the health concerns of adolescent girls sending unsolicited emails to a teenage girls' magazine. METHOD: We conducted a content analysis of 1000 systematic randomly selected unsolicited emails submitted to the health column of an Australian adolescent girls' magazine over a 6-month period. RESULTS: Three main foci of concern were identified: Context of Concern, Health Issue of Concern and Advice Sought for Concern. Within Health Issue of Concern, there were five categories: body (47.5%), sex (31.9%), relationship (14.7%), mind (4.7%) and violence and/or safety (1.2%). Concerns within the body and sex categories ranged enormously, but frequently expressed intimate descriptions of anatomy, feelings, sexual practices and relationships. Many concerns occurred in the context of adolescents' relationships with others. The proportion of concerns about physical or psychological symptoms or health issues commonly associated with the adolescent age group (such as health risk behaviours, mental health, pregnancy and sexually transmitted infections) was relatively small. CONCLUSIONS: GPs and other health professionals might engage more readily with adolescent patients with a deeper understanding of the concerns that adolescents have about their bodies, relationships and overall health. Seemingly 'trivial' issues, such as normal puberty, could be used as discussion triggers in health consultations to help alleviate anxiety and build rapport. PMID- 19289551 TI - Myocardial glucose and lactate metabolism during rest and atrial pacing in humans. AB - There is minimal in vivo data in humans evaluating myocardial substrate utilization during increased heart work. This study was performed to determine the balance of myocardial glucose and lactate metabolism during rest and increased heart work induced by atrial pacing in seven healthy men and women (age, 49.7 +/- 3.9 years; body mass index, 23.4 +/- 1.1 kg m(-2), maximum oxygen consumption, 35.5 +/- 3.0 ml kg(-1) min(-1), ejection fraction, 68 +/- 3%). After 3 days of dietary control, catheters were placed in coronary sinus, femoral arterial and venous, and peripheral venous blood vessels. Subjects received a primed continuous infusion of [3,3,3-(2)H]lactate and [6,6-(2)H]glucose throughout the study. Arterial and coronary sinus blood sampling and measurements of coronary sinus blood flow were made during rest and atrial pacing at approximately 111 beats min(-1). Myocardial oxygen consumption increased (P = 0.04) from rest to atrial pacing. Net glucose uptake increased (P = 0.04) from rest to atrial pacing with unchanged fractional extraction (rest: 9.1 +/- 2.7%, atrial pacing 9.8 +/- 2.9%). The percentage of whole body glucose disposal from myocardial uptake also increased from rest to atrial pacing. Isotopically measured lactate uptake also increased significantly from rest to atrial pacing with no significant differences in fractional extraction. The myocardium released lactate throughout the experiment, which increased significantly from rest and atrial pacing (P < 0.05). The heart accounted for a significantly greater percentage of whole body lactate disposal during atrial pacing (15.0 +/- 4.4%) compared to rest (4.9 +/- 0.9%, P = 0.03). These data suggest: (1) in the absence of ischaemia the myocardium is constantly taking up and releasing lactate at rest which increases during atrial pacing, and (2) when arterial substrate delivery is unchanged, increased myocardial work is accomplished with similar proportions of glucose and lactate utilization in healthy humans in vivo. PMID- 19289553 TI - Intralobular pulmonary lymphatic distribution in normal human lung using D2-40 antipodoplanin immunostaining. AB - It has been assumed for a long time that except for limited areas close to respiratory bronchioles or their satellite arteries, there is no evidence of lymphatic vessels deep in the pulmonary lobule. An immunohistochemical study using the D2-40 monoclonal antibody was performed on normal pulmonary samples obtained from surgical specimens, with particular attention to the intralobular distribution of lymphatic vessels. This study demonstrated the presence of lymphatics not only in the connective tissue surrounding the respiratory bronchioles but also associated with intralobular arterioles and/or small veins even less than 50 mum in diameter. A few interlobular lymphatic vessels with a diameter ranging from 10 mum to 20 mum were also observed further away, in interalveolar walls. In conclusion, this study, using the D2-40 monoclonal antibody, demonstrated the presence of small lymphatic channels within the normal human pulmonary lobules, emerging from interalveolar interstitium, and around small blood vessels constituting the paraalveolar lymphatics. This thin intralobular lymphatic network may play a key pathophysiological role in a wide variety of alveolar and interstitial lung diseases and requires further investigation. PMID- 19289555 TI - Reflections on management. PMID- 19289554 TI - Image analysis algorithms for immunohistochemical assessment of cell death events and fibrosis in tissue sections. AB - Cell death is of broad physiological and pathological importance, making quantification of biochemical events associated with cell demise a high priority for experimental pathology. Fibrosis is a common consequence of tissue injury involving necrotic cell death. Using tissue specimens from experimental mouse models of traumatic brain injury, cardiac fibrosis, and cancer, as well as human tumor specimens assembled in tissue microarray (TMA) format, we undertook computer-assisted quantification of specific immunohistochemical and histological parameters that characterize processes associated with cell death. In this study, we demonstrated the utility of image analysis algorithms for color deconvolution, colocalization, and nuclear morphometry to characterize cell death events in tissue specimens: (a) subjected to immunostaining for detecting cleaved caspase 3, cleaved poly(ADP-ribose)-polymerase, cleaved lamin-A, phosphorylated histone H2AX, and Bcl-2; (b) analyzed by terminal deoxyribonucleotidyl transferase mediated dUTP nick end labeling assay to detect DNA fragmentation; and (c) evaluated with Masson's trichrome staining. We developed novel algorithm-based scoring methods and validated them using TMAs as a high-throughput format. The proposed computer-assisted scoring methods for digital images by brightfield microscopy permit linear quantification of immunohistochemical and histochemical stainings. Examples are provided of digital image analysis performed in automated or semiautomated fashion for successful quantification of molecular events associated with cell death in tissue sections. PMID- 19289556 TI - "Can I drive, doctor?" LEAN thinking may help us answer the question. AB - The loss of a driving licence can have a devastating effect on an individual's domestic life and work. Driving is a complex composite function comprising a number of other complex composite functions, and it is difficult to predict driving ability just from the traditional neurological examination. We propose the acronym LEAN as an aide-memoire to help structure a suggested driving section in the routine neurological clerking, and to help unpack the concept of "fitness" to drive: Licence status, Eligibility (to drive), Ability (to drive) and Notification requirements (to the licensing authorities), now and in the future; cover most of the important issues and may help ensure that people get the best advice. If there are concerns about someone's driving ability, with or without vehicle adaptations, an assessment in a Disabled Drivers Assessment Unit is recommended. PMID- 19289557 TI - Not all morning headaches are due to brain tumours. AB - Headaches causing early morning waking, or headaches which are more prominent on waking, always raise the suspicion of raised intracranial pressure, and hence the need for prompt evaluation to exclude the diagnosis of a brain tumour (particularly if they are associated with vomiting and papilloedema). However, there are many other much more common causes of "morning headache", both primary and secondary. As ever, history taking is key to the diagnosis. Attention to the possibility of analgesic medication overuse is particularly pertinent, but other treatable conditions such as depression and epilepsy must not be overlooked. PMID- 19289558 TI - When a patient with epilepsy complains about poor memory. AB - Patients with epilepsy often complain of "poor memory". The first step in managing this complaint is a clinical evaluation to define and, if possible, quantify the problem. The memory difficulty may be entirely unconnected to the epilepsy. But if the two appear to be linked, establish whether the memory problem is due to the seizures themselves, the pathology that underlies the seizures, their treatment, or their psychological sequelae such as anxiety or depression. Further management depends on the cause, while practical advice on the amelioration of poor memory can be useful. PMID- 19289559 TI - A difficult case solved at autopsy: memory loss, behavioural change and seizures. PMID- 19289560 TI - James Paget's median nerve compression (Putnam's acroparaesthesia). PMID- 19289561 TI - Fahr's disease. PMID- 19289562 TI - The paroxysmal dyskinesias. AB - The paroxysmal dyskinesias are a challenging group of movement disorders characterised by painless dystonic and/or choreiform movements. Lack of familiarity with their features and a normal neurological examination between attacks frequently cause diagnostic delays, or even the diagnosis of a non organic disorder. They are classified by their mode of triggering, and also by the duration and frequency of attacks, effectiveness of medication, and any associated syndromes. Four subtypes are recognised: paroxysmal kinesigenic dyskinesia induced by sudden movement; paroxysmal non-kinesigenic dyskinesia precipitated by for instance alcohol or caffeine; paroxysmal exercise-induced dyskinesia triggered by longer lasting activity; and paroxysmal hypnogenic dyskinesia occurring during sleep. Here we will summarise the characteristics of the subtypes, discuss the differential diagnosis, genetic aspects and pathophysiology, and give practical advice on the diagnostic work-up and treatment. PMID- 19289563 TI - Recurrent choking in Parkinson's disease. PMID- 19289564 TI - Northern Ireland. PMID- 19289565 TI - Multiple sclerosis. PMID- 19289567 TI - Efficacy of ipamorelin, a novel ghrelin mimetic, in a rodent model of postoperative ileus. AB - Ghrelin and ghrelin mimetics stimulate appetite and enhance gastric motility. The present study investigates whether ipamorelin, a selective growth hormone secretagogue and agonist of the ghrelin receptor, would accelerate gastrointestinal transit and ameliorate the symptoms in a rodent model of postoperative ileus (POI). Fasted male rats were subjected to laparotomy and intestinal manipulation. At the end of surgery, a dye marker was infused in the proximal colon to evaluate postsurgical colonic transit time, which was the time to the first bowel movement. In addition, fecal pellet output, food intake, and body weight were monitored regularly for 48 h. Ipamorelin (0.01-1 mg/kg), growth hormone-releasing peptide (GHRP)-6 (20 microg/kg), or vehicle (saline) were administered via intravenous bolus infusion after a single dosing or a 2-day repetitive dosing regimen (four doses a day at 3-h intervals). Compared with the vehicle, a single dose of ipamorelin (1 mg/kg) or GHRP-6 (20 microg/kg) decreased the time to the first bowel movement but had no effect on cumulative fecal output, food intake, or body weight gain measured 48 h after the surgery. In contrast, repetitive dosing of ipamorelin (0.1 or 1 mg/kg) significantly increased the cumulative fecal pellet output, food intake, and body weight gain. The results suggest that postsurgical intravenous infusions of ipamorelin may ameliorate the symptoms in patients with POI. PMID- 19289568 TI - Selective inhibitors of CYP2J2 related to terfenadine exhibit strong activity against human cancers in vitro and in vivo. AB - The cytochrome P450 epoxygenase, CYP2J2, converts arachidonic acid to four regioisomeric epoxyeicosatrienoic acids (EETs). We found recently that this enzyme is dramatically up-regulated in a variety of established human carcinoma cell lines and in human cancerous tissue and promotes the neoplastic phenotype. In the present study, we tested the hypothesis that specific inhibitors of CYP2J2 related to the drug terfenadine are effective antitumor agents. Four of these inhibitors (compounds 4, 5, 11, and 26) were tested for effectiveness in vitro and in vivo. In Tca-8113 cells, the CYP2J2 inhibitors decreased EET production by approximately 60%, whereas they had no effect on CYP2J2 mRNA or protein expression. Compound 26 inhibited the proliferation of human tumor cells, reduced their ability to adhere, invade, and migrate, and attenuated activation of epithelial growth factor receptor signal and kinases and phosphatidylinositol 3 kinase/Akt pathways. Inhibition of CYP2J2 also significantly potentiated human tumor cell apoptosis and caused a corresponding increase in caspase-3 activity and change in expression of apoptosis-related proteins Bax and Bcl-2. In murine xenograft models using MDA-MB-435 cells, treatment with compound 26 significantly repressed tumor growth, decreased lung metastasis, and was associated with increased expression of the anticancer genes CD82 and nm23, without causing toxicity. These data suggest that CYP2J2 inhibitors hold significant promise for use in treatment of neoplastic diseases. PMID- 19289569 TI - Activation and modulation of concatemeric GABA-A receptors expressed in human embryonic kidney cells. AB - We have employed whole-cell and single-channel electrophysiology to examine the kinetic and pharmacological properties of GABA-A receptors consisting of gamma2L beta2-alpha1 and beta2-alpha1 subunit concatemeric constructs expressed in human embryonic kidney cells. Concatemeric receptors activated by GABA exhibited the same single-channel conductance, channel opening rate constant, and basic open- and closed-time properties as receptors containing free subunits. However, the whole-cell GABA dose-response and the single-channel effective opening rate curves were shifted to higher GABA concentrations, suggesting that the concatemeric receptors have a lower affinity to GABA. Pharmacological tests demonstrated that the concatemeric receptors were potentiated by pentobarbital, diazepam, and the neurosteroid (3alpha,5alpha)-3-hydroxypregnan-20-one (3alpha5alphaP), and were insensitive to Zn(2+). Selective introduction of the alpha1Q241L mutation, previously shown to abolish alpha1beta2gamma2L channel potentiation by neurosteroids, into one of the two concatemeric constructs had a relatively small effect on receptor activation by GABA or macroscopic potentiation by the neurosteroid 3alpha5alphaP. Single-channel measurements showed that the kinetic mechanism of action of the steroid is unchanged when the mutation is introduced to the gamma2L-beta2-alpha1 concatemer. We infer that a single wild-type alpha subunit is capable of mediating the full set of kinetic effects in the presence of steroids. Introduction of the alpha1Q241W mutation, previously shown to mimic the effect of the steroid on alpha1beta2gamma2L channels, selectively into either concatemeric construct altered the mode of activity elicited by P4S, but the presence of mutations in both alpha subunits was required to affect open-time distributions. The data indicate that the alpha1Q241W mutation acts as a partial steroid modulator. PMID- 19289570 TI - Met909 plays a key role in the activation of the progesterone receptor and also in the high potency of 13-ethyl progestins. AB - Many progestins have been developed for use in contraception, menopausal hormone therapy, and treatment of gynecological diseases. They are derived from either progesterone or testosterone, and they act by binding to the progesterone receptor (PR), a hormone-inducible transcription factor belonging to the nuclear receptor superfamily. Unlike mineralocorticoid, glucocorticoid, and androgen receptors, the steroid-receptor contacts that trigger the switch of the ligand binding domain from an inactive to an active conformation have not yet been identified for the PR. With this aim, we solved the crystal structure of the ligand-binding domain of the human PR complexed with levonorgestrel, a potent testosterone-derived progestin characterized by a 13-ethyl substituent. Via mutagenesis analysis and functional studies, we identified Met909 of the helix 12 as the key residue for PR activation by both testosterone- and progesterone derived progestins with a 13-methyl or a 13-ethyl substituent. We also showed that Asn719 contributes to PR activation by testosterone-derived progestins only, and that Met759 and Met909 are responsible for the high potency of 19 norprogestins and of 13-ethyl progestins, respectively. Our findings provide a structural guideline for the rational synthesis of potent PR agonist and antagonist ligands that could have therapeutic uses in women's health. PMID- 19289571 TI - mu-Opioid receptor cell surface expression is regulated by its direct interaction with Ribophorin I. AB - The trafficking of the mu-opioid receptor (MOR), a member of the rhodopsin G protein-coupled receptor (GPCR) family, can be regulated by interaction with multiple cellular proteins. To determine the proteins involved in receptor trafficking, using the targeted proteomic approach and mass spectrometry analysis, we have identified that Ribophorin I (RPNI), a component of the oligosaccharide transferase complex, could directly interact with MOR. RPNI can be shown to participate in MOR export by the intracellular retention of the receptor after small interfering RNA knockdown of endogenous RPNI. Overexpression of RPNI rescued the surface expression of the MOR 344KFCTR348 deletion mutant independent of calnexin. Furthermore, RPNI regulation of MOR trafficking is dependent on the glycosylation state of the receptor, as reflected by the inability of overexpression of RPNI to affect the trafficking of the N glycosylation-deficient mutants, or GPCRs that have minimal glycosylation sites. Hence, this novel RPNI chaperone activity is a consequence of N-glycosylation dependent direct interaction with MOR. PMID- 19289572 TI - The analysis of desensitizing CNGA1 channels reveals molecular interactions essential for normal gating. AB - The pore region of cyclic nucleotide-gated (CNG) channels acts as the channel gate. Therefore, events occurring in the cyclic nucleotide-binding (CNB) domain must be coupled to the movements of the pore walls. When Glu363 in the pore region, Leu356 and Thr355 in the P helix, and Phe380 in the upper portion of the S6 helix are mutated into an alanine, gating is impaired: mutant channels E363A, L356A, T355A, and F380A desensitize in the presence of a constant cGMP concentration, contrary to what can be observed in wild-type (WT) CNGA1 channels. Similarly to C-type inactivation of K(+) channels, desensitization in these mutant channels is associated with rearrangements of residues in the outer vestibule. In the desensitized state, Thr364 residues in different subunits become closer and Pro366 becomes more accessible to extracellular reagents. Desensitization is also observed in the mutant channel L356C, but not in the double-mutant channel L356C+F380C. Mutant channels L356F and F380K did not express, but cGMP-gated currents with a normal gating were observed in the double mutant channels L356F+F380L and L356D+F380K. Experiments with tandem constructs with L356C, F380C, and L356C+F380C and WT channels indicate that the interaction between Leu356 and Phe380 is within the same subunit. These results show that Leu356 forms a hydrophobic interaction with Phe380, coupling the P helix with S6, whereas Glu363 could interact with Thr355, coupling the pore wall to the P helix. These interactions are essential for normal gating and underlie the transduction between the CNB domain and the pore. PMID- 19289573 TI - Elevations of intracellular calcium reflect normal voltage-dependent behavior, and not constitutive activity, of voltage-dependent calcium channels in gastrointestinal and vascular smooth muscle. AB - In smooth muscle, the gating of dihydropyridine-sensitive Ca(2+) channels may either be stochastic and voltage dependent or coordinated among channels and constitutively active. Each form of gating has been proposed to be largely responsible for Ca(2+) influx and determining the bulk average cytoplasmic Ca(2+) concentration. Here, the contribution of voltage-dependent and constitutively active channel behavior to Ca(2+) signaling has been studied in voltage-clamped single vascular and gastrointestinal smooth muscle cells using wide-field epifluorescence with near simultaneous total internal reflection fluorescence microscopy. Depolarization (-70 to +10 mV) activated a dihydropyridine-sensitive voltage-dependent Ca(2+) current (I(Ca)) and evoked a rise in [Ca(2+)] in each of the subplasma membrane space and bulk cytoplasm. In various regions of the bulk cytoplasm the [Ca(2+)] increase ([Ca(2+)](c)) was approximately uniform, whereas that of the subplasma membrane space ([Ca(2+)](PM)) had a wide range of amplitudes and time courses. The variations that occurred in the subplasma membrane space presumably reflected an uneven distribution of active Ca(2+) channels (clusters) across the sarcolemma, and their activation appeared consistent with normal voltage-dependent behavior. Indeed, in the present study, dihydropyridine-sensitive Ca(2+) channels were not normally constitutively active. The repetitive localized [Ca(2+)](PM) rises ("persistent Ca(2+) sparklets") that characterize constitutively active channels were observed rarely (2 of 306 cells). Neither did dihydropyridine-sensitive constitutively active Ca(2+) channels regulate the bulk average [Ca(2+)](c). A dihydropyridine blocker of Ca(2+) channels, nimodipine, which blocked I(Ca) and accompanying [Ca(2+)](c) rise, reduced neither the resting bulk average [Ca(2+)](c) (at -70 mV) nor the rise in [Ca(2+)](c), which accompanied an increased electrochemical driving force on the ion by hyperpolarization (-130 mV). Activation of protein kinase C with indolactam-V did not induce constitutive channel activity. Thus, although voltage dependent Ca(2+) channels appear clustered in certain regions of the plasma membrane, constitutive activity is unlikely to play a major role in [Ca(2+)](c) regulation. The stochastic, voltage-dependent activity of the channel provides the major mechanism to generate rises in [Ca(2+)]. PMID- 19289574 TI - SLC26A9 is a constitutively active, CFTR-regulated anion conductance in human bronchial epithelia. AB - Human bronchial epithelial (HBE) cells exhibit constitutive anion secretion that is absent in cells from cystic fibrosis (CF) patients. The identity of this conductance is unknown, but SLC26A9, a member of the SLC26 family of CF transmembrane conductance regulator (CFTR)-interacting transporters, is found in the human airway and exhibits chloride channel behavior. We sought differences in the properties of SLC26A9 and CFTR expressed in HEK 293 (HEK) cells as a fingerprint to identify HBE apical anion conductances. HEK cells expressing SLC26A9 displayed a constitutive chloride current that was inhibited by the CFTR blocker GlyH-101 (71 +/- 4%, 50 microM) and exhibited a near-linear current voltage (I-V) relation during block, while GlyH-101-inhibited wild-type (wt)CFTR exhibited a strong inward-rectified (IR) I-V relation. We tested polarized HBE cells endogenously expressing either wt or DeltaF508-CFTR for similar activity. After electrical isolation of the apical membrane using basolateral alpha-toxin permeabilization, wtCFTR monolayers displayed constitutive chloride currents that were inhibited by GlyH-101 (68 +/- 6%) while maintaining a near-linear I-V relation. In the absence of blocker, the addition of forskolin stimulated a current increase having a linear I-V; GlyH-101 blocked 69 +/- 7% of the current and shifted the I-V relation IR, consistent with CFTR activation. HEK cells coexpressing SLC26A9 and wtCFTR displayed similar properties, as well as forskolin-stimulated currents that exceeded the sum of those in cells separately expressing SLC26A9 or wtCFTR, and an I-V relation during GlyH-101 inhibition that was moderately IR, indicating that SLC26A9 contributed to the stimulated current. HBE cells from CF patients expressed SLC26A9 mRNA, but no constitutive chloride currents. HEK cells coexpressing SLC26A9 with DeltaF508-CFTR also failed to exhibit SLC26A9 current. We conclude that SLC26A9 functions as an anion conductance in the apical membranes of HBE cells, it contributes to transepithelial chloride currents under basal and cAMP/protein kinase A stimulated conditions, and its activity in HBE cells requires functional CFTR. PMID- 19289576 TI - Thymic progenitor homing and lymphocyte homeostasis are linked via S1P-controlled expression of thymic P-selectin/CCL25. AB - Thymic T cell progenitor (TCP) importation is a periodic, gated event that is dependent on the expression of functional P-selectin ligands on TCPs. Occupancy of intrathymic TCP niches is believed to negatively regulate TCP importation, but the nature of this feedback mechanism is not yet resolved. We show that P selectin and CCL25 are periodically expressed in the thymus and are essential parts of the thymic gate-keeping mechanism. Periodicity of thymic TCP receptivity and the size of the earliest intrathymic TCP pool were dependent on the presence of functional P-selectin ligand on TCPs. Furthermore, we show that the numbers of peripheral blood lymphocytes directly affected thymic P-selectin expression and TCP receptivity. We identified sphingosine-1-phosphate (S1P) as one feedback signal that could mediate influence of the peripheral lymphocyte pool on thymic TCP receptivity. Our findings suggest a model whereby thymic TCP importation is controlled by both early thymic niche occupancy and the peripheral lymphocyte pool via S1P. PMID- 19289577 TI - Normally occurring NKG2D+CD4+ T cells are immunosuppressive and inversely correlated with disease activity in juvenile-onset lupus. AB - The NKG2D receptor stimulates natural killer cell and T cell responses upon engagement of ligands associated with malignancies and certain autoimmune diseases. However, conditions of persistent NKG2D ligand expression can lead to immunosuppression. In cancer patients, tumor expression and shedding of the MHC class I-related chain A (MICA) ligand of NKG2D drives proliferative expansions of NKG2D(+)CD4(+) T cells that produce interleukin-10 (IL-10) and transforming growth factor-beta, as well as Fas ligand, which inhibits bystander T cell proliferation in vitro. Here, we show that increased frequencies of functionally equivalent NKG2D(+)CD4(+) T cells are inversely correlated with disease activity in juvenile-onset systemic lupus erythematosus (SLE), suggesting that these T cells may have regulatory effects. The NKG2D(+)CD4(+) T cells correspond to a normally occurring small CD4 T cell subset that is autoreactive, primed to produce IL-10, and clearly distinct from proinflammatory and cytolytic CD4 T cells with cytokine-induced NKG2D expression that occur in rheumatoid arthritis and Crohn's disease. As classical regulatory T cell functions are typically impaired in SLE, it may be clinically significant that the immunosuppressive NKG2D(+)CD4(+) T cells appear functionally uncompromised in this disease. PMID- 19289578 TI - Crops and climate change: progress, trends, and challenges in simulating impacts and informing adaptation. AB - Assessments of the relationships between crop productivity and climate change rely upon a combination of modelling and measurement. As part of this review, this relationship is discussed in the context of crop and climate simulation. Methods for linking these two types of models are reviewed, with a primary focus on large-area crop modelling techniques. Recent progress in simulating the impacts of climate change on crops is presented, and the application of these methods to the exploration of adaptation options is discussed. Specific advances include ensemble simulations and improved understanding of biophysical processes. Finally, the challenges associated with impacts and adaptation research are discussed. It is argued that the generation of knowledge for policy and adaptation should be based not only on syntheses of published studies, but also on a more synergistic and holistic research framework that includes: (i) reliable quantification of uncertainty; (ii) techniques for combining diverse modelling approaches and observations that focus on fundamental processes; and (iii) judicious choice and calibration of models, including simulation at appropriate levels of complexity that accounts for the principal drivers of crop productivity, which may well include both biophysical and socio-economic factors. It is argued that such a framework will lead to reliable methods for linking simulation to real-world adaptation options, thus making practical use of the huge global effort to understand and predict climate change. PMID- 19289579 TI - Occupational asthma and the paper recycling industry. AB - BACKGROUND: Occupational disease linked to the paper recycling industry has not been well documented. No previously confirmed formal diagnosis of occupational asthma (OA) caused by hydroxylamine has been made. METHODS: We have assessed and performed occupational assessment of eight workers involved in this industry. Two of these were later diagnosed with OA and are reported here. RESULTS: Both workers developed their respiratory symptoms within 2 years of the first use of the chemical hydroxylamine as part of the 'de-inking' process. Hydroxylamine was used as a substitute for glutaraldehyde on risk grounds, although no prior cases of OA had been found. The two workers had worked at the same plant for 11 and 20 years, respectively. Both gave histories of work-related wheeze, shortness of breath and cough. Both cases performed OASYS peak flow records over a 3-week period and had OASYS II index of 2.85 and 2.67, respectively. Both were redeployed on site to non-exposed areas and subsequently demonstrated improvement in bronchial reactivity. Case 2 subsequently consented to and underwent a blinded, placebo-controlled occupational challenge using hydroxylamine demonstrating a significant isolated late asthmatic response. CONCLUSIONS: We believe that these are the first two confirmed cases of OA caused by hydroxylamine in the paper recycling industry. PMID- 19289580 TI - Quality in pathology: of Deming, the Edsel, and Henry Ford. PMID- 19289581 TI - Disclosing harmful pathology errors to patients. PMID- 19289582 TI - The Henry Ford Production System: reduction of surgical pathology in-process misidentification defects by bar code-specified work process standardization. AB - Misidentification defects are a potential patient safety issue in medicine, including in the surgical pathology laboratory. In addressing the Joint Commission's national patient safety goal of accurate patient and specimen identification, we focused our lens internally on our own laboratory processes, with measurement tools designed to identify potential misidentification defects and their root causes. Based on this knowledge, aligned with our lean work culture in the Henry Ford Production System, we redesigned our surgical pathology laboratory workflow with simplified connections and pathways reinforced by a bar code technology innovation to specify and standardize work processes. We also adopted just-in-time prestain slide labeling with solvent-impervious, bar-coded slide labels at the microtome station, eliminating the loop-back pathway of poststain, batch slide matching, and labeling with adhesive paper labels. These changes have enabled us to dramatically reduce the overall misidentification case rate by approximately 62% with an approximate 95% reduction in the more common histologic slide misidentification defects while increasing technical throughput at the histology microtomy station by 125%. PMID- 19289583 TI - EGFR and K-ras mutations along the spectrum of pulmonary epithelial tumors of the lung and elaboration of a combined clinicopathologic and molecular scoring system to predict clinical responsiveness to EGFR inhibitors. AB - We tested 418 neoplasms along the whole spectrum of primary lung tumor histotypes for epidermal growth factor receptor (EGFR) and K-ras mutations. Clinicopathologic data from 154 patients undergoing treatment with EGFR tyrosine kinase inhibitors (TKIs) were retrospectively studied. A scoring system assigning a score for each positive or negative characteristic (+1, female sex, nonsmoking status, adenocarcinoma histotype, Asian ethnicity, and EGFR mutation; -1, current smoker and K-ras mutation; and 0, male sex, ex-smoker, nonadenocarcinoma histotype, and no mutations) was elaborated and tested with EGFR-TKI response. Salivary gland-type, mucin-rich, and neuroendocrine tumors do not harbor EGFR mutations. A subset of nonmucinous adenocarcinomas, not necessarily of the bronchioloalveolar type, is related to EGFR mutations. Three probability groups significantly correlating with response to EGFR-TKIs were identified. Of note, the addition of molecular results did not significantly change the predictive value obtained by the combination of clinicopathologic characteristics alone in this scoring system. K-ras mutations, significantly associated with the mucin secreting type of adenocarcinoma, consistently predict lack of response in white patients. PMID- 19289584 TI - Chromogenic in situ hybridization is a reliable method for detecting HER2 gene status in breast cancer: a multicenter study using conventional scoring criteria and the new ASCO/CAP recommendations. AB - Chromogenic in situ hybridization (CISH) has shown the potential to replace fluorescence in situ hybridization (FISH) to determine HER2 gene status. To validate the reliability of CISH, we used 226 consecutive breast carcinomas from 2 institutions and tested CISH and FISH on the same tumor set simultaneously at different test sites. Besides manufacturers' scoring criteria, the new American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) guidelines were used to interpret HER2 status. The concordance between CISH and FISH for positive and negative results was 98.5% at site A and 98.6% at site B using the manufacturers' criteria, and 99.0% at site A and 99.1% at site B using the ASCO/CAP criteria. Reproducibility of CISH results was more than 98.0% among 3 sites using the manufacturers' criteria and 100.0% between 2 sites using the ASCO/CAP criteria. Our results confirm that CISH is reliable for HER2 testing per ASCO/CAP guidelines. PMID- 19289585 TI - Peritumoral activated hepatic stellate cells predict poor clinical outcome in hepatocellular carcinoma after curative resection. AB - The inflammatory components of the liver remnant after hepatocellular carcinoma (HCC) resection are of prognostic importance. We evaluated prognostic potential of peritumoral activated hepatic stellate cells (HSCs) in 130 HCC cases. The messenger RNA (mRNA) levels of the functional genes in HSCs (ie, seprase, osteonectin, and tenascin-C), quantitated by real-time quantitative polymerase chain reaction, and the density of peritumoral Foxp3+ T-regulatory cells (Tregs) and CD68+ macrophages (MPhi), assessed immunohistochemically in tissue microarray sections, were positively correlated with the density of peritumoral activated HSCs. The density (P= .007 for recurrence-free survival [RFS] and P=.021 for overall survival [OS]) and functional genes (seprase, P= .001 for RFS; osteonectin, P= .007 for RFS and P=.021 for OS) of peritumoral activated HSCs independently contributed to high recurrence or death rates, as did peritumoral Tregs or MPhi. Moreover, peritumoral HSCs were related to more early recurrences. It is important to note that the density of peritumoral activated HSCs, in combination with seprase and osteonectin mRNA or density of Tregs and MPhi, might predict prognoses more effectively. PMID- 19289586 TI - Clusterin expression correlates with stage and presence of large cells in mycosis fungoides. AB - Clusterin expression is common in systemic and cutaneous anaplastic large cell lymphoma (ALCL). Mycosis fungoides (MF) in large cell transformation can resemble ALCL. In this study, we immunohistochemically assessed for clusterin in 97 skin biopsy specimens, including 70 MF cases and 27 other cutaneous neoplasms including ALCL, peripheral T-cell lymphoma unspecified (PTCL), and lymphomatoid papulosis (LyP). Clusterin was positive in 36 (51%) of 70 cases of MF and correlated with clinical stage in 68 cases: 3 of 21 stage I, 11 of 20 stage II, and 23 of 27 stage III/IV. Clusterin expression also correlated with type of skin lesion (3/19 patch, 13/28 plaque, and 20/23 tumor/erythroderma) and number of large cells (6/30 small cell, 12/18 with increased large cells, and 18/22 with large cell transformation). Clusterin expression was not specific for MF as it also was positive in 3 of 3 cases of LyP, 2 of 2 systemic ALCL cases involving skin, 7 of 16 cutaneous ALCLs, and 1 of 6 PTCLs. PMID- 19289587 TI - Usefulness of EMA, GLUT-1, and XIAP for the cytologic diagnosis of malignant mesothelioma in body cavity fluids. AB - We compared the effectiveness of epithelial membrane antigen (EMA) with 2 newly described markers, X-linked inhibitor of apoptosis protein (XIAP) and an isoform of glucose transporter (GLUT-1), in the distinction between malignant mesothelioma (MM) and benign effusion (BE) in body cavity fluids. Immunohistochemical studies were performed on cell block sections from 35 cases of histologically confirmed MM and 38 BEs, using antibodies to EMA, XIAP, GLUT-1 (GLUT-1m, monoclonal; GLUT-1p, polyclonal). Results were graded based on the percentage of cells staining: negative (0%), 1+ (<25%), 2+ (25%-49%), 3+ (50% 74%), and 4+ (75%-100%). The performance of each marker was compared using receiver operating characteristic curve analysis. EMA demonstrated the best accuracy, with an area under the curve of 0.91 as compared with XIAP (0.67), GLUT 1m (0.74), and GLUT-1p (0.80). Based on these findings, EMA is a better marker than XIAP or GLUT-1 for the diagnosis of MM. PMID- 19289588 TI - Evaluation of an abbreviated protocol for cytomegalovirus pp65 antigenemia testing. AB - The cytomegalovirus antigenemia assay remains a useful tool for monitoring reactivation among transplant recipients. In this study, we compared protocols using direct lysis of small-volume, whole blood (WB) samples vs peripheral blood leukocyte (PBL)-enriched fractions. Of 363 evaluable samples, 51 (14.0%) were positive by one or both methods. Sensitivity, specificity, and negative and positive predictive values were similar (76%, 99%, 96%, and 95% vs 71%, 100%, 96%, and 100% for the WB and PBL methods, respectively). Stratification of qualitative results by WBC count revealed comparable detection rates by each method, although the total number of positive results from leukopenic samples was significantly lower than from nonleukopenic samples (P= .04). Correlation between quantitative results was high, yet the degree of clinical agreement was suboptimal. We conclude that the small-volume, WB lysis method yields results statistically comparable to that of a PBL fractionation method but with fewer technical steps and less complexity. PMID- 19289589 TI - Chromogenic media vs real-time PCR for nasal surveillance of methicillin resistant Staphylococcus aureus: impact on detection of MRSA-positive persons. AB - Surveillance for methicillin-resistant Staphylococcus aureus (MRSA) colonization can be an important element for infection control programs when managing a multidrug-resistant pathogen such as MRSA. The sensitivity and speed of laboratory testing affects the proportion of appropriate isolation days captured, which determines the success or failure of a MRSA control program. Chromogenic culture, CHROMagar MRSA (BBL, Becton Dickinson, Sparks, MD) and MRSASelect (Bio Rad, Hercules, CA), with and without broth enrichment and real-time polymerase chain reaction (PCR; BD GeneOhm MRSA, BD Diagnostics, San Diego, CA), were compared and found to have a wide range of sensitivities (78.5%-98.2%), specificities (91.6%-100.0%), and turnaround times (2-72 hours). Real-time PCR provided the most rapid results and demonstrated the highest sensitivity followed by broth-enriched culture and then direct plating for MRSA detection in nasal swabs. There was no substantial difference in the labor required for any of the 3 approaches. PMID- 19289590 TI - Analysis of bacterial detection in whole blood-derived platelets by quantitative glucose testing at a university medical center. AB - After the March 2004 implementation of American Association of Blood Banks standards regarding platelet bacterial detection, we began quantitative glucose screening of whole blood-derived platelets (WB-P). The glucose level was measured immediately before component release--often storage day 4 or 5--using the Glucometer SureStep Flexx Meter (LifeScan, Milpitas, CA), with a positive cutoff of less than 500 mg/dL; failing units were cultured and not transfused. During 29 months (March 1, 2004-July 31, 2006) 93,073 units of WB-P were tested. Initially, 929 units (0.998%) screened positively. Bacterial growth was culture-confirmed in 6 units, for a bacterial contamination incidence of 0.006% and a true-positive rate of 6.4/100,000. Three additional culture-confirmed contamination cases were detected in transfused units causing febrile nonhemolytic reactions, for a false negative rate of 3.2/100,000. Our overall contamination prevalence was 9.6/100,000 units of platelets transfused, lower than ordinarily cited, and showed a false-negative rate remarkably congruent to that of culture: 3.2/100,000. A low-sensitivity screening test applied late in platelet shelf-life can be comparable to culture in preventing bacterial-related morbidity. PMID- 19289591 TI - Laboratory assessment of factor VIII inhibitor titer: the North American Specialized Coagulation Laboratory Association experience. AB - Quantification of inhibitory antibodies against infused factor VIII (FVIII) has an important role in the management of patients with hemophilia A. This article summarizes results from the largest North American FVIII inhibitor proficiency testing challenge conducted to date. Test samples, 4 negative and 4 positive (1-3 Bethesda units [BU]/mL), were distributed by the ECAT Foundation in conjunction with the North American Specialized Coagulation Laboratory Association and analyzed by 38 to 42 laboratories in 2006 and 2007. Whereas laboratories were able to distinguish between the absence and presence of low-titer FVIII inhibitors, the intralaboratory coefficient of variation was high (30%-42%) for inhibitor-positive samples, and the definition of lower detection limits of the assay was variable (0-1 BU/mL). Most laboratories performed the Bethesda assay with commercially supplied buffered normal pooled plasma in a 1:1 mix with patient plasma. These data provide information for the development of consensus guidelines to improve FVIII inhibitor quantification. PMID- 19289592 TI - Issues in neonatal cellular analysis. AB - Hematologic values in neonates differ significantly from those in older children and adults. Quantitative and qualitative differences are present as a reflection of the developmental changes during fetal hematopoiesis and, so, correlate with gestational age. At birth, the hemoglobin, mean corpuscular volume, and WBC counts of term newborns are significantly higher than those of older children and adults, and in preterm neonates the differences are even more pronounced. This review explores these differences and the major factors that account for them from the hematology laboratory standpoint. After a discussion of the developmental hematopoiesis and normal hematologic values in term and preterm neonates, important preanalytic factors, such as limited blood availability, effect of sampling site, and violent crying, and analytic interferences are examined. Finally, the review addresses resulting challenges in interpretation of hematologic test results in term and preterm neonates, especially issues surrounding neonatal reference intervals and critical value reporting, and suggests possible solutions. PMID- 19289593 TI - Frequent expression of CD99 in anaplastic large cell lymphoma: a clinicopathologic and immunohistochemical study of 160 cases. AB - Originally described as a diagnostically useful marker for Ewing sarcoma, CD99 immunoreactivity has also been documented in a variety of other tumors, including hematopoietic neoplasms. By using conventional paraffin immunoperoxidase staining and tissue microarrays, we retrospectively investigated CD99 expression in a series of 160 anaplastic large cell lymphoma (ALCL) cases. Of the 160 cases, 103 (64.4%) were positive for CD99. The distribution of CD99 positivity was similar for nodal (66/103 [64.1%]), extranodal, (21/32 [66%]), and primary cutaneous lesions (16/25 [64%]). CD99 expression was present in 96 (64.4%) of 149 of the common type, 4 (80%) of 5 of the small cell variant, and 3 (50%) of 6 of the lymphohistiocytic variant cases. CD99 expression was slightly more frequent in anaplastic large cell lymphoma kinase (ALK)+ cases compared with ALK- cases (43/54 [80%] vs 44/81 [54%]). With 2 exceptions, ALK+ ALCL was seen only in patients younger than 41 years. We conclude that CD99 is frequently expressed in ALCL, with a slightly increased frequency in the younger age ALK+ cases. Nodal and extranodal ALCL should be considered in the differential diagnosis when a CD99+ neoplasm is encountered. PMID- 19289594 TI - Evidence that stainable bone marrow iron following parenteral iron therapy does not correlate with serum iron studies and may not represent readily available storage iron. AB - We recently reported that parenteral iron therapy is associated with a characteristic pattern of iron staining on bone marrow aspirate smears. We now present clinical information from 6 patients who received parenteral iron and, at one or more points in follow-up, were found to have low or borderline low serum ferritin levels and/or serum iron levels, even though marrow aspirate smears revealed abundant stainable iron in the pattern characteristic of prior parenteral iron therapy. We conclude that stainable iron seen in this pattern does not correlate with serum iron studies and may not represent functionally available storage iron. This pattern of iron staining should not be used as evidence to withhold further iron therapy in patients who otherwise continue to have features of iron deficiency anemia. PMID- 19289595 TI - Immunophenotypic analysis of CD103+ B-lymphoproliferative disorders: hairy cell leukemia and its mimics. AB - CD103 is characteristically expressed in hairy cell leukemia (HCL), a B lymphoproliferative disorder highly responsive to treatment with purine analogs. Other CD103+ diseases are rare and do not respond well to the same therapy, including HCL variant (HCLv) and splenic marginal zone B-cell lymphoma (SMZL) variants. We analyzed 215 cases of CD103+ B-lymphoproliferative disorders to further delineate their immunophenotypic features. Flow cytometric analysis revealed that 78.6% of all cases expressed CD25 and CD103, characteristic of classical HCL. Cases analyzed immunohistochemically were also invariably positive for annexin-A1; a subset coexpressed CD10 (33/169 [19.5%]) or BCL1 (26/65 [36.9%]). In contrast, 21.4% of cases lacked CD25, a subset of which was analyzed and was invariably negative for annexin-A1, CD10, and BCL1. The CD25- cases had variable morphologic features ranging from HCLv and SMZL to prolymphocytic leukemia and diffuse large B-cell lymphoma. Clinically, patients with CD25- disease tended to be older (P= .001), typically had leukocytosis (P= .014), and did not respond well to cladribine or pentostatin. We suggest categorizing CD103+ B-lymphoproliferative disorders into 2 groups. While HCL coexpresses CD25 and annexin-A1, diseases lacking CD25 and annexin-A1 behave clinically differently and can be separated from HCL on diagnosis. PMID- 19289596 TI - Evolution of the Caenorhabditis elegans genome. AB - A fundamental problem in genome biology is to elucidate the evolutionary forces responsible for generating nonrandom patterns of genome organization. As the first metazoan to benefit from full-genome sequencing, Caenorhabditis elegans has been at the forefront of research in this area. Studies of genomic patterns, and their evolutionary underpinnings, continue to be augmented by the recent push to obtain additional full-genome sequences of related Caenorhabditis taxa. In the near future, we expect to see major advances with the onset of whole-genome resequencing of multiple wild individuals of the same species. In this review, we synthesize many of the important insights to date in our understanding of genome organization and function that derive from the evolutionary principles made explicit by theoretical population genetics and molecular evolution and highlight fertile areas for future research on unanswered questions in C. elegans genome evolution. We call attention to the need for C. elegans researchers to generate and critically assess nonadaptive hypotheses for genomic and developmental patterns, in addition to adaptive scenarios. We also emphasize the potential importance of evolution in the gonochoristic (female and male) ancestors of the androdioecious (hermaphrodite and male) C. elegans as the source for many of its genomic and developmental patterns. PMID- 19289597 TI - Evidence for homologous recombination in intracellular chemosynthetic clam symbionts. AB - Homologous recombination is a fundamental mechanism for the genetic diversification of free-living bacteria. However, recombination may be limited in endosymbiotic bacteria, as these taxa are locked into an intracellular niche and may rarely encounter sources of foreign DNA. This study tested the hypothesis that vertically transmitted endosymbionts of deep-sea clams (Bivalvia: Vesicomyidae) show little or no evidence of recombination. Phylogenetic analysis of 13 loci distributed across the genomes of 14 vesicomyid symbionts revealed multiple, well-supported inconsistencies among gene tree topologies, and maximum likelihood-based tests rejected a hypothesis of shared evolutionary history (linkage) among loci. Further, multiple statistical methods confirmed the presence of recombination by detecting intragenic breakpoints in two symbiont loci. Recombination may be confined to a subset of vesicomyid symbionts, as some clades showed high levels of genomic stability, whereas others showed clear patterns of homologous exchange. Notably, a mosaic genome is present in symB, a symbiont lineage shown to have been acquired laterally (i.e., nonvertically) by Vesicomya sp. JdF clams. The majority of loci analyzed here supported a tight sister clustering of symB with the symbiont of a host species from the Mid Atlantic Ridge, whereas others placed symB in a clade with symA, the dominant phylotype of V. sp. JdF clams. This result raises the hypothesis that lateral symbiont transfer between hosts may facilitate recombination by bringing divergent symbiont lineages into contact. Together, the data show that homologous recombination contributes to the diversification of vesicomyid clam symbionts, despite the intracellular lifestyle of these bacteria. PMID- 19289598 TI - mRNA retrotransposition coupled with 5' inversion as a possible source of new genes. AB - Human long interspersed nuclear element-1 (L1) occupies one-sixth of our genome and has contributed to genome evolution in various ways. Approximately 10% of human L1 copies are composed of two L1 segments; the 5' segment and 3' segment are in head-to-head (i.e., 5'-inverted) orientation. Besides mediating their own retrotransposition, L1 has the ability to mobilize mRNA "in trans," and the number of retrotransposed mRNA sequences (retrocopies) is estimated to be >6,000. In this study, we identified 48 human-specific retrocopies and 95 chimpanzee specific retrocopies by comparing the human and chimpanzee genomes. Among these retrocopies, 12 were 5'-inverted. The characteristics of these 5'-inverted retrocopies were similar to those of 5'-inverted L1 copies, indicating that the 5' inversion is generated by the same mechanism. With these findings, we examined the possibility that 5' inversion of the retrocopy generates a new gene that codes for a peptide with a different N terminus. We identified several potential 5'-inverted retrogenes, including those of thymopoietin beta (TMPO) and eukaryotic translation initiation factor 3 subunit 5 (EIF3F). The most interesting candidate was the 5'-inverted retrocopy of small nuclear ribonucleoprotein polypeptide N (SNRPN). This retrocopy was transcribed in the reverse orientation in several organs, had multiple transcript variants, and encoded a protein containing a peptide fragment derived from the N-terminal portion of SNRPN. Our results suggest that mRNA retrotransposition coupled with 5' inversion may be a mechanism to generate new genes distinct from parental genes. PMID- 19289599 TI - Duplications and expression of DIVARICATA-like genes in dipsacales. AB - The genetics underlying flower symmetry shifts between radial and bilateral symmetry has been intensively studied in the model Antirrhinum majus. Understanding the conservation or diversification of this genetic pathway in other plants is of special interest in understanding angiosperm evolution and ecology. Evidence from Antirrhinum indicates that TCP and MYB transcription factors, especially CYCLOIDEA (CYC), DICHOTOMA (DICH), DIVARICATA (DIV), and RADIALIS (RAD) play a role in specifying dorsal identity (CYC, DICH, and RAD) and ventral identity (DIV) in the corolla and androecium of monosymmetric (bilateral) flowers. Previous data indicate that the ECE clade of TCP genes (including CYC and DICH) underwent two duplication events around the diversification of the core eudicots. In this study, we examined the duplication events within Dipsacales, which contains both radially and bilaterally symmetrical flowered species. Additionally, we report here the phylogenetic relationships of the DIV-like genes across core eudicots. Like TCP genes, we found three core eudicot clades of DIV like genes, with duplications occurring around the diversification of the core eudicots, which we name DIV1, DIV2, and DIV3. The Antirrhinum genes, DIVARICATA and its sister DVL1, fall into the DIV1 clade. We also found additional duplications within these clades in Dipsacales. Specifically, the Caprifoliaceae (bilaterally symmetrical clade) duplicated independently in each of the three core eudicot DIV clades. Using reverse transcription polymerase chain reaction (rtPCR), we showed that most of these copies are expressed across floral tissues in the Dipsacales species Heptacodium miconioides. One copy, DipsDIV1A (orthologous to DIV and DVL1), was expressed in a dorsal-ventral pattern. DipsDIV1A was expressed only in petal tissue, in both dorsal and ventral regions but was lacking from lateral petals. We argue that this suggests that DipsDIV1A may be expressed in a similar pattern to DIV in Antirrhinum, suggesting a broad conservation of this pathway. Finally, DIV contains a large intron near the beginning of the second MYB domain, which shows promise as a highly variable molecular marker for phylogenetic studies. PMID- 19289600 TI - A genomewide comparison of population structure at STRPs and nearby SNPs in humans. AB - Patterns of population structure provide insights into evolutionary processes and help identify groups of individuals for genotype-phenotype association studies. With increasing availability of polymorphic molecular markers across genomes, the examination of population structure using large numbers of unlinked loci has become a common practice in evolutionary biology and human genetics. The two classes of molecular variation most widely used for this purpose, short tandem repeat polymorphisms (STRPs) and single-nucleotide polymorphisms (SNPs), differ in mutational properties expected to affect population structure. To measure the relative ability of these loci to describe population structure, we compared diversity at neighboring STRPs and SNPs from 720 genomic regions in the four populations that comprise the Human HapMap. Comparing loci from the same genomic regions allowed us to focus on the contribution of mutational differences (rather than variation in genealogical history) to disparities in population structure between STRPs and SNPs. Relative to average values for SNPs from the same regions, STRPs had lower F(st), but higher G(st)' and I(n) values. STRP-SNP correlations in population structure across genomic regions were statistically significant but weak in magnitude. Separate analyses by repeat type showed that these correlations were driven primarily by tetranucleotide and trinucleotide STRPs; measures of population structure at dinucleotides and SNPs were not significantly correlated. Pairwise comparisons among populations revealed effects of divergence time on differences in population structure between STRPs and SNPs. Collectively, these results confirm that individual STRPs can provide more information about population structure than individual SNPs, but suggest that the difference in structure at STRPs and SNPs depends on local genealogical history. Our study motivates theoretical comparisons of population structure at loci with different mutational properties. PMID- 19289601 TI - TLR4/MyD88/PI3K interactions regulate TLR4 signaling. AB - TLRs activate immune responses by sensing microbial structures such as bacterial LPS, viral RNA, and endogenous "danger" molecules released by damaged host cells. MyD88 is an adapter protein that mediates signal transduction for most TLRs and leads to activation of NF-kappaB and MAPKs and production of proinflammatory cytokines. TLR4-mediated signaling also leads to rapid activation of PI3K, one of a family of kinases involved in regulation of cell growth, apoptosis, and motility. LPS stimulates phosphorylation of Akt, a downstream target of PI3K, in wild-type (WT) mouse macrophages. LPS-induced phosphorylation of Akt serine 473 was blunted in MyD88(-/-) macrophages and was completely TLR4-dependent. MyD88 and p85 were shown previously to co-immunoprecipitate, and a YXXM motif within the Toll-IL-1 resistance (TIR) domain of MyD88 was suggested to be important for this interaction. To test this hypothesis, we compared expressed MyD88 variants with mutations within the YXXM motif or lacking the TIR domain or death domain and measured their capacities to bind PI3K p85, MyD88, and TLR4 by co immunoprecipitation analyses. The YXXM --> YXXA mutant MyD88 bound more strongly to p85, TLR4, and WT MyD88 than the other variants, yet was significantly less active than WT MyD88, suggesting that sustained interaction of MyD88/PI3K with the TLR4 intracellular "signaling platform" negatively regulates signaling. We propose a hypothetical model in which sustained PI3K activity at the membrane limits the availability of the PI3K substrate, thereby negatively regulating signaling. PMID- 19289602 TI - Does it mean anything if your own name is wrong in your published paper? AB - Journals instruct authors to proofread their accepted manuscripts before signing them off for publication and expect them to submit errata to correct any mistakes identified thereafter. Herein, I examine papers with obvious errors in the author name list. I queried PubMed for papers under common Greek forenames looking for citations where author surnames and forenames are swapped. I identified 113 such papers from 101 journals. Author names are corrected with errata only in 20, after a median of 6.5 mo. Time to name correction is shorter for journals with impact factor above the median (P=0.015). To further explore this suggested association of apparent author sloppiness with journal impact, I use as controls all errata published between 1996 and 2008 in 5 top-cited general medical journals (New England Journal of Medicine, JAMA, The Lancet, Annals of Internal Medicine, and British Medical Journal); 3.4% of the latter contain name corrections (vs. 18.1% of all errata in the 101 journals during the same period, P<10(-6)). Egregious errors may be markers of cursory if any proofreading and, therefore, markers for additional unidentified inaccuracies. In addition, I wonder whether authors may be as reluctant to rectify other, nonobvious (yet potentially consequential) mistakes after a paper's publication. PMID- 19289603 TI - Imatinib attenuates skeletal muscle dystrophy in mdx mice. AB - Duchenne-Meryon muscular dystrophy (DMD) is the most common and lethal genetic muscle disease. Ameliorating muscle necrosis, inflammation, and fibrosis represents an important therapeutic approach for DMD. Imatinib, an antineoplastic agent, demonstrated antiinflammatory and antifibrotic effects in liver, kidney, lung, and skin of various animal models. This study tested antiinflammatory and antifibrotic effects of imatinib in mdx mice, a DMD mouse model. We treated mdx mice with intraperitoneal injections of imatinib at the peak of limb muscle inflammation and the onset of diaphragm fibrosis. Controls received PBS vehicle or were left untreated. Muscle necrosis, inflammation, fibrosis, and function were evaluated by measuring serum CK activity, endomysial CD45 immunoreactive inflammation area, endomysial collagen III deposition, and hind limb grip strength. Phosphorylation of the tyrosine kinase targets of imatinib was assessed by Western blot in diaphragm tissue and in primary cultures of peritoneal macrophages and skeletal muscle fibroblasts. Imatinib markedly reduced muscle necrosis, inflammation, and fibrosis, and significantly improved hind limb grip strength in mdx mice. Reduced clinical disease was accompanied by inhibition of c abl and PDGFR phosphorylation and suppression of TNF-alpha and IL-1beta expression. Imatinib therapy for DMD may hold promise for ameliorating muscle necrosis, inflammation, and fibrosis by inhibiting c-abl and PDGFR signaling pathways and downstream inflammatory cytokine and fibrotic gene expression. PMID- 19289604 TI - Folding mechanism of the C-terminal domain of nucleophosmin: residual structure in the denatured state and its pathophysiological significance. AB - Nucleophosmin (NPM1) is a ubiquitously expressed protein and is one of the most abundant proteins found in the nucleolus. Naturally occurring mutations in the C terminal domain of nucleophosmin (Cter-NPM1) are found in approximately 30% of patients with acute myeloid leukemia (AML). These mutations cause changes at the C terminus of NPM1 that lead to denaturation of the protein, a critical factor in determining aberrant translocation of NPM1 to the cytosol. Hence, this protein system represents an ideal candidate to investigate the relations between folding and unfolding and disease. Here we report the characterization of the folding and unfolding kinetics of Cter-NPM1. Data reveal that this small helical domain folds via a compact denatured state, displaying a malleable residual structure. Moreover, analysis of folding rate constants measured under different experimental conditions suggests that the existence of a preorganized structure in the denatured state accelerates folding, implying a native-like residual structure. Because a major feature of Cter-NPM1 mutants responsible for AML is a reduction in stability of the protein and thus prevalence of a denatured state even under physiological conditions, our findings may pave the way to further studies with the aim of designing chemicals capable of interacting with the "pathological" mutants to stabilize the native conformation. PMID- 19289605 TI - Identification and characterization of a complete carnitine biosynthesis pathway in Candida albicans. AB - Carnitine is an essential metabolite that enables intracellular transport of fatty acids and acetyl units. Here we show that the yeast Candida albicans can synthesize carnitine de novo, and we identify the 4 genes of the pathway. Null mutants of orf19.4316 (trimethyllysine dioxygenase), orf19.6306 (trimethylaminobutyraldehyde dehydrogenase), and orf19.7131 (butyrobetaine dioxygenase) lacked their respective enzymatic activities and were unable to utilize fatty acids, acetate, or ethanol as a sole carbon source, in accordance with the strict requirement for carnitine-mediated transport under these growth conditions. The second enzyme of carnitine biosynthesis, hydroxytrimethyllysine aldolase, is encoded by orf19.6305, a member of the threonine aldolase (TA) family in C. albicans. A strain lacking orf19.6305 showed strongly reduced growth on fatty acids and was unable to utilize either acetate or ethanol, but TA activity was unaffected. Growth of the null mutants on nonfermentable carbon sources is restored only by carnitine biosynthesis intermediates after the predicted enzymatic block in the pathway, which provides independent evidence for a specific defect in carnitine biosynthesis for each of the mutants. In conclusion, we have genetically characterized a complete carnitine biosynthesis pathway in C. albicans and show that a TA family member is mainly involved in the aldolytic cleavage of hydroxytrimethyllysine in vivo. PMID- 19289606 TI - Binding of heat shock protein 70 to extracellular phosphatidylserine promotes killing of normoxic and hypoxic tumor cells. AB - Hypoxia is well known to limit curability of tumors by ionizing radiation. Here, we show that hypoxia treatment of tumor cells causes coexpression of heat shock protein 70 (Hsp70) and phosphatidylserine (PS) on the cell surface. Colocalization of Hsp70 and PS, as determined by confocal microscopy, also occurs when exogenous FITC-labeled Hsp70 protein is added to normoxic and hypoxic tumor cells. Moreover, the interaction of Hsp70 with PS was demonstrated in artificial unilamellar phosphatidylcholine/ phosphatidylserine (PC/PS) liposomes at the physiological ratio of 8/2. Indeed, the Hsp70-liposome interaction gradually increased with elevating PS molar ratios (8/2 > or = 7/3 < 5/5 < 4/6 < 3/7 < 2/8). In contrast, only a weak Hsp70 interaction was detected in phosphatidylcholine/phosphatidylglycerol (PC/PG) liposomes, thus demonstrating that the interaction was not a charge-related effect. The interaction of Hsp70 with surface PS significantly reduces clonogenic cell survival in normoxic (EC(50) of Hsp70=85 microg/ml) and hypoxic (EC(50) of Hsp70=55 microg/ml) tumor cells. The radiation-induced tumor cell killing was significantly enhanced by the addition of Hsp70 protein (50 microg/ml). Since apoptosis was not significantly enhanced in normoxic and hypoxic tumor cells by the addition of Hsp70, we hypothesize that the Hsp70 protein-induced reduction in clonogenic cell survival might be through necrosis rather than apoptosis. PMID- 19289607 TI - Accumulation of tau induced in neurites by microglial proinflammatory mediators. AB - Aggregated fibrillary microtubule-associated protein tau is the major component of neurofibrillary tangles in Alzheimer's disease. The exact molecular mechanism of tau aggregation is unknown. Microglial cell activation and migration toward amyloid-beta plaques precede the appearance of dysmorphic neurites and formation of neurofibrillary tangles. Here, we analyzed the accumulation of tau at a distance range of expected spontaneous aggregation by fluorescence lifetime-based Forster resonance energy transfer in cultured primary murine neurons cotransfected with the human tau gene tagged to the green fluorescent protein variants Citrine (tau-Citrine) and Cerulean (tau-Cerulean). No spontaneous accumulation of cotransfected tau-Citrine and tau-Cerulean was detected in untreated neurons. Coculture of neurons with activated microglia induced aggregation of tau in neurites. Treatment of neurons with tumor necrosis factor alpha (TNF-alpha) stimulated reactive oxygen species generation and resulted in the accumulation of tau-Citrine and tau-Cerulean in neurites, which was inhibited by neutralization of TNF and the free radical inhibitor 6-hydroxy-2,5,7,8 tetramethylchromane-2-carboxylic acid (Trolox). These data demonstrate that activated microglia and the microglial-derived proinflammatory cytokine TNF can induce accumulation of the aggregation-prone tau molecules in neurites via reactive oxygen species. PMID- 19289608 TI - Treating IIA/B seminoma with combination carboplatin and radiotherapy. PMID- 19289609 TI - Study of newspaper reports regarding clinical trials in Japan. PMID- 19289610 TI - Health behaviors influence cancer survival. PMID- 19289612 TI - Chemotherapy alone for laryngeal preservation--is it possible? PMID- 19289611 TI - High-risk populations identified in Childhood Cancer Survivor Study investigations: implications for risk-based surveillance. AB - Childhood cancer survivors often experience complications related to cancer and its treatment that may adversely affect quality of life and increase the risk of premature death. The purpose of this manuscript is to review how data derived from Childhood Cancer Survivor Study (CCSS) investigations have facilitated identification of childhood cancer survivor populations at high risk for specific organ toxicity and secondary carcinogenesis and how this has informed clinical screening practices. Articles previously published that used the resource of the CCSS to identify risk factors for specific organ toxicity and subsequent cancers were reviewed and results summarized. CCSS investigations have characterized specific groups to be at highest risk of morbidity related to endocrine and reproductive dysfunction, pulmonary toxicity, cerebrovascular injury, neurologic and neurosensory sequelae, and subsequent neoplasms. Factors influencing risk for specific outcomes related to the individual survivor (eg, sex, race/ethnicity, age at diagnosis, attained age), sociodemographic status (eg, education, household income, health insurance) and cancer history (eg, diagnosis, treatment, time from diagnosis) have been consistently identified. These CCSS investigations that clarify risk for treatment complications related to specific treatment modalities, cumulative dose exposures, and sociodemographic factors identify profiles of survivors at high risk for cancer-related morbidity who deserve heightened surveillance to optimize outcomes after treatment for childhood cancer. PMID- 19289613 TI - Continuing trastuzumab beyond progression. PMID- 19289614 TI - Predictors of postoperative quality of life after esophagectomy for cancer. AB - PURPOSE: To identify factors predictive of health-related quality of life (HRQL) 6 months after surgery for esophageal cancer. PATIENTS AND METHODS: A Swedish nationwide population-based study of patients undergoing esophagectomy for cancer was undertaken between 2001 and 2005. Clinical details (sex, age, body mass index [BMI], comorbidity) and tumor details (histology, stage, and location) were prospectively recorded and validated. HRQL questionnaires (European Organisation for Research and Treatment of Cancer Quality of Life Questionnaires C30 and OES18) were used to assess HRQL 6 months postoperatively. HRQL outcome was categorized as good or poor by collapsing the four response categories. Logistic regression models adjusting for potential confounding factors were used to assess the association between preoperative patient and tumor characteristics and HRQL outcome 6 months after surgery. RESULTS: A total of 355 patients (77% of eligible) responded to the HRQL questionnaires. Age, sex, and BMI showed no associations with HRQL 6 months after surgery, but patients with comorbidity, a more advanced tumor stage (III to IV), or a tumor located in the middle or upper esophagus had an increased risk of poor HRQL. Patients with adenocarcinoma had a lower risk of poor HRQL than patients with squamous cell carcinoma. CONCLUSION: Knowledge of risk factors for poor postoperative HRQL may be relevant for clinical decision making. More research to see whether these factors are further predictive of poor HRQL in long-term survivors is needed. PMID- 19289615 TI - Effect of HPV-associated p16INK4A expression on response to radiotherapy and survival in squamous cell carcinoma of the head and neck. AB - PURPOSE: A subset of head and neck cancers is associated with the human papillomavirus (HPV). Viral infection is closely correlated with expression of p16(INK4A) in these tumors. We evaluated p16(INK4A) as a prognostic marker of treatment response and survival in a well-defined and prospectively collected cohort of patients treated solely with conventional radiotherapy in the Danish Head and Neck Cancer Group (DAHANCA) 5 trial. PATIENTS AND METHODS: Immunohistochemical expression of p16(INK4A) was analyzed in pretreatment paraffin-embedded tumor blocks from 156 patients treated with conventional primary radiotherapy alone. The influence of p16(INK4A) status on locoregional tumor control, disease-specific survival, and overall survival after radiotherapy was evaluated. RESULTS: p16(INK4A) positivity was found in 35 tumors (22%). Tumor positivity for p16(INK4A) was significantly correlated with improved locoregional tumor control (5-year actuarial values 58% v 28%; P = .0005), improved disease specific survival (72% v 34%; P = .0006), and improved overall survival (62% v 26%; P = .0003). In multivariate analysis, p16(INK4A) remained a strong independent prognostic factor for locoregional failure (hazard ratio [HR], 0.35; 95% CI, 0.19 to 0.64), disease-specific death (HR, 0.36; 95% CI, 0.20 to 0.64), and overall death (HR, 0.44; 95% CI, 0.28 to 0.68). CONCLUSION: Expression of p16(INK4A) has a major impact on treatment response and survival in patients with head and neck cancer treated with conventional radiotherapy. PMID- 19289616 TI - Prognostic factors for survival after salvage reirradiation of head and neck cancer. AB - PURPOSE: Patients who develop recurrent or new primary head and neck cancer in a previously irradiated site have poor prognosis. Reirradiation is a treatment option, although it is associated with substantial toxicities. We investigated potential prognostic factors, including comorbidity and pre-existing organ dysfunction, for survival after reirradiation. METHODS: Institutional electronic records of patients treated with reirradiation between January 1998 and 2008 were reviewed. Comorbidity was assessed by Charlson index and Adult Comorbidity Evaluation-27 (ACE-27) grading. Organ dysfunction was defined as feeding tube dependency, functioning tracheostomy, or soft tissue defect. RESULTS: There were 103 patients, including 46 patients who underwent salvage surgery before reirradiation. Median progression-free and overall survivals were 12.1 months (95% CI, 9.7 to 16.6) and 19.3 months (95% CI, 13.9 to 29.9), respectively. Significant comorbidity was present in 36% of patients by Charlson index and 24% by ACE-27. Baseline organ dysfunction was present in 37% of patients. Median overall survivals were 5.5 months among those with both organ dysfunction and comorbidity per Charlson index, and 4.9 months per ACE-27, compared with 59.6 and 44.2 months, respectively, among the patients with neither organ dysfunction nor comorbidity (P < .001 and < .001). Other independent prognostic factors were interval from previous radiation, recurrent tumor stage, tumor bulk at reirradiation, and reirradiation dose. A nomogram to predict the probability of death within 24 months after reirradiation was developed (concordance index = 0.75). CONCLUSION: Comorbidity and pre-existing organ dysfunction are among several important prognostic factors for patients undergoing reirradiation. For those with both comorbidity and organ dysfunction, reirradiation largely serves as a palliative therapy. PMID- 19289617 TI - Identification of a predictive biomarker for hematologic toxicities of gemcitabine. AB - PURPOSE: Gemcitabine monotherapy is the current standard for patients with advanced pancreatic cancer, but the occurrence of severe neutropenia and thrombocytopenia can sometimes be life threatening. This study aimed to discover a new diagnostic method for predicting the hematologic toxicities of gemcitabine. PATIENTS AND METHODS: Using quantitative mass spectrometry (MS), we compared the baseline plasma proteomes of 25 patients who had developed severe hematologic adverse events (grade 3 to 4 neutropenia and/or grade 2 to 4 thrombocytopenia) within the first two cycles of gemcitabine with those of 22 patients who had not (grade 0). RESULTS: We identified 757 peptide peaks whose intensities were significantly different (P < .001, Welch t test) among a total of 60,888. The MS peak with the highest statistical significance (P = .0000282) was revealed to be derived from haptoglobin by tandem MS. A scoring system (nomogram) based on the values of haptoglobin, haptoglobin phenotype, neutrophil count, platelet count, and body-surface area was constructed to estimate the risk of hematologic adverse events (grade 3 to 4 neutropenia and/or grade 2 to 4 thrombocytopenia) with an area under curve value of 0.782 in a cohort of 166 patients with pancreatic cancer. Predictive ability of the system was confirmed in two independent validation cohorts consisting of 87 and 52 patients with area under the curve values of 0.655 and 0.747, respectively. CONCLUSION: Although the precise mechanism responsible for the correlation of haptoglobin with the future onset of hematologic toxicities remains to be clarified, our prediction model seems to have high practical utility for tailoring the treatment of patients receiving gemcitabine. PMID- 19289618 TI - Baseline C-reactive protein is associated with incident cancer and survival in patients with cancer. AB - PURPOSE: We tested the hypothesis that baseline plasma levels of C-reactive protein (CRP) are associated with risk of incident cancer in the general population and early death in patients with cancer. PATIENTS AND METHODS: A total of 10,408 individuals from the Danish general population who had CRP measured at baseline were observed for up to 16 years; 1,624 developed cancer, and of these, 998 patients died during follow-up. Follow-up was 100% complete. We excluded individuals with a cancer diagnosis at baseline. RESULTS: Baseline CRP levels more than 3 versus less than 1 mg/L were associated with multifactorially adjusted hazard ratios of 1.3 (95% CI, 1.0 to 1.6) for cancer of any type, 2.2 (95% CI, 1.0 to 4.6) for lung cancer, 1.9 (95% CI, 0.8 to 4.6) for colorectal cancer, and 0.7 (95% CI, 0.4 to 1.4) for breast cancer. Corresponding hazard ratios for the highest versus the lowest quintile of baseline CRP levels were 1.3 (95% CI, 1.0 to 1.6), 2.1 (95% CI, 1.2 to 3.8), 1.7 (95% CI, 0.8 to 3.2), and 0.9 (95% CI, 0.5 to 1.7), respectively. Multifactorially adjusted hazard ratios for early death in patients with cancer were 1.8 (95% CI, 1.2 to 2.7) for CRP more than 3 versus less than 1 mg/L and 1.4 (95% CI, 1.1 to 1.7) for the highest versus the lowest quintile. Elevated CRP levels were associated with early death in patients with cancer having localized disease, but not in those with metastases (interaction; P = .03). CONCLUSION: Elevated levels of CRP in cancer free individuals are associated with increased risk of cancer of any type, of lung cancer, and possibly of colorectal cancer. Moreover, elevated levels of baseline CRP associate with early death after a diagnosis of any cancer, particularly in patients without metastases. PMID- 19289619 TI - Trastuzumab beyond progression in human epidermal growth factor receptor 2 positive advanced breast cancer: a german breast group 26/breast international group 03-05 study. AB - PURPOSE: Trastuzumab shows clinical activity in human epidermal growth factor receptor 2 (HER-2)-positive early and advanced breast cancer. In the German Breast Group 26/Breast International Group 03-05 trial, we investigated if trastuzumab treatment should be continued beyond progression. METHODS: Patients with HER-2-positive breast cancer that progresses during treatment with trastuzumab were randomly assigned to receive capecitabine (2,500 mg/m(2) body surface area on days 1 through 14 [1,250 mg/m(2) semi-daily]) alone or with continuation of trastuzumab (6 mg/kg body weight) in 3-week cycles. The primary end point was time to progression. RESULTS: We randomly assigned 78 patients to capecitabine and 78 patients to capecitabine plus trastuzumab. Sixty-five events and 38 deaths in the capecitabine group and 62 events and 33 deaths in the capecitabine-plus-trastuzumab group occurred during 15.6 months of follow-up. Median times to progression were 5.6 months in the capecitabine group and 8.2 months in the capecitabine-plus-trastuzumab group with an unadjusted hazard ratio of 0.69 (95% CI, 0.48 to 0.97; two-sided log-rank P = .0338). Overall survival rates were 20.4 months (95% CI, 17.8 to 24.7) in the capecitabine group and 25.5 months (95% CI, 19.0 to 30.7) in the capecitabine-plus-trastuzumab group (P = .257). Overall response rates were 27.0% with capecitabine and 48.1% with capecitabine plus trastuzumab (odds ratio, 2.50; P = .0115). Continuation of trastuzumab beyond progression was not associated with increased toxicity. CONCLUSION: Continuation of trastuzumab plus capecitabine showed a significant improvement in overall response and time to progression compared with capecitabine alone in women with HER-2-positive breast cancer who experienced progression during trastuzumab treatment. PMID- 19289620 TI - Phase II study of picoplatin as second-line therapy for patients with small-cell lung cancer. AB - PURPOSE: This study was designed to confirm the efficacy and safety of picoplatin, a cisplatin analog designed to overcome platinum resistance, in patients with small-cell lung cancer (SCLC) with platinum-refractory/-resistant disease. PATIENTS AND METHODS: All patients received intravenous picoplatin 150 mg/m(2) every 3 weeks. Tumor response, progression-free survival, and overall survival were evaluated. Adverse events were assessed for frequency, severity, and relationship to treatment. Quality of life was assessed with the Lung Cancer Symptom Scale instrument. RESULTS: Seventy-seven patients were treated with picoplatin (median number of cycles, two; range one to 10). Three patients (4%) had a partial response, 33 (43%) had stable disease (four of these were unconfirmed partial responses), 36 (47%) had progressive disease, and five were not assessable for response. Median progression-free survival was 9.1 weeks (95% CI, 7.0 to 12.1 weeks). Median overall survival was 26.9 weeks (95% CI, 21.1 to 33.4). The most common grade 3 and 4 toxicities were thrombocytopenia (48%), neutropenia (25%), and anemia (20%). The most commonly reported adverse events of any severity included thrombocytopenia (64%), anemia (49%), neutropenia (39%), nausea (27%), fatigue (16%), and dyspnea (16%). No severe neurotoxicity or nephrotoxicity were observed. There were no treatment-related deaths. CONCLUSION: Picoplatin demonstrated clinical efficacy in platinum-refractory SCLC. The major toxicity was hematologic. These results warrant further evaluation in this patient population. PMID- 19289621 TI - Qualitative and quantitative assessment of taste and smell changes in patients undergoing chemotherapy for breast cancer or gynecologic malignancies. AB - PURPOSE: Smell and taste changes during chemotherapy are significant complaints of cancer patients. Loss of olfactory/gustatory function can lead to malnutrition, weight loss, and possibly a prolonged morbidity of chemotherapy induced adverse effects, decreased quality of life, poor compliance, and even decreased therapy response. This prospective study comprehensively investigated, to our knowledge for the first time, smell and taste changes in a cohort of 87 patients undergoing chemotherapy for breast cancer or gynecologic malignancies. PATIENTS AND METHODS: Olfactory function was tested using Sniffin' Sticks (Burghart; Wedel, Germany) and gustatory function was tested using taste strips before, during, and immediately and 3 months after chemotherapy. RESULTS: Olfactory and gustatory function significantly decreased during chemotherapy and recovered almost completely 3 months after chemotherapy. Scores of odor thresholds were affected more than those of discrimination or identification. The olfactory function of older patients was affected more than that of younger patients. There was no difference in the olfactory function during chemotherapy with respect to the chemotherapeutic agent or initial diagnosis (breast or ovarian cancer). Regarding taste, scores of salty taste were affected more than scores of sweet, sour, or bitter taste. The gustatory function did not differ significantly during chemotherapy with respect to age or diagnosis but did differ with respect to the chemotherapeutic agent. Taxane-based chemotherapy caused the most severe disorders. CONCLUSION: Chemotherapy has a significant but transient effect on olfactory and gustatory function, possibly causing reduced appetite, a low energy intake, and weight loss. Additional spices and flavoring may compensate for this diminished chemosensory function, enhancing patient compliance and quality of life. PMID- 19289622 TI - Microsatellite instability, mismatch repair deficiency, and BRAF mutation in treatment-resistant germ cell tumors. AB - PURPOSE: Mismatch repair (MMR) deficiency and microsatellite instability (MSI) are associated with cisplatin resistance in human germ cell tumors (GCTs). BRAF mutation (V600E) is found in MSI colorectal cancers. The role of RAS/RAF pathway mutations in GCT treatment response is unknown. PATIENTS AND METHODS: Two patient cohorts were investigated: 100 control GCTs (50 seminomas and 50 nonseminomas) and 35 cisplatin-based chemotherapy-resistant GCTs. MMR proteins were analyzed by immunohistochemistry, and eight microsatellite loci were examined for MSI. Tumors were assessed for specific BRAF and KRAS mutations. RESULTS: Resistant tumors showed a higher incidence of MSI than controls: 26% versus 0% in two or more loci (P < .0001). All resistant tumors were wild-type KRAS, and two controls (2%) contained a KRAS mutation. There was a significantly higher incidence of BRAF V600E mutation in resistant tumors compared with controls: 26% versus 1% (P < .0001). BRAF mutations were highly correlated with MSI (P = .006), and MSI and mutated BRAF were correlated with weak or absent staining for hMLH1 (P = .017 and P = .008). Low or absent staining of hMLH1 was correlated with promoter hypermethylation (P < .001). Tumors lacking expression of hMLH1 or MSH6 were significantly more frequent in resistant GCTs than in controls (P = .001 and 0.0036, respectively). Within the subgroup of resistant tumors, patients with MSI showed a trend to longer progression-free survival (P = .068). CONCLUSION: We report for the first time a correlation between a gene mutation--BRAF V600E--and cisplatin resistance in nonseminomatous GCTs. Furthermore, a correlation between MMR deficiency, MSI, and treatment failure is confirmed. PMID- 19289623 TI - Randomized phase II study of gefitinib compared with placebo in chemotherapy naive patients with advanced non-small-cell lung cancer and poor performance status. AB - PURPOSE: To compare gefitinib with placebo in chemotherapy naive patients with advanced non-small-cell lung cancer (NSCLC) and poor performance status. PATIENTS AND METHODS: NSCLC patients (chemotherapy naive, WHO performance status 2 or 3; unfit for chemotherapy; stage IIIB/IV) were randomly assigned to gefitinib (250 mg/d) plus best supportive care (BSC; n = 100) or placebo plus BSC (n = 101). The primary end point was progression-free survival (PFS). Secondary end points included overall survival (OS), objective response rate (ORR), quality of life (QOL), pulmonary symptom improvement (PSI), and safety. Correlation of gefitinib efficacy with EGFR gene copy number (fluorescent in situ hybridization [FISH]) was explored. RESULTS: Hazard ratios (HRs; gefitinib:placebo) were 0.82 (95% CI, 0.60 to 1.12; P = .217) for PFS and 0.84 (95% CI, 0.62 to 1.15; P = .272) for OS. As expected for this patient population, OS for both arms was poor, at about 3 months. ORRs were 6.0% (gefitinib) and 1.0% (placebo). QOL and PSI rates were 21.1% and 28.3% (gefitinib) and 20.0% and 28.3% (placebo), respectively. In EGFR FISH-positive patients (n = 32), HRs were 0.29 (95% CI, 0.11 to 0.73) for PFS and 0.44 (95% CI, 0.17 to 1.12) for OS. No unexpected adverse events occurred. CONCLUSION: There was no statistically significant difference in PFS, OS, and ORRs after treatment with gefitinib or placebo, in the overall population; improvements in QOL and symptoms were similar in both groups. Tolerability profile of gefitinib was consistent with previous studies. PFS was statistically significantly improved for gefitinib-treated patients with EGFR FISH-positive tumors. PMID- 19289624 TI - Incidence, treatment costs, and complications of lymphedema after breast cancer among women of working age: a 2-year follow-up study. AB - PURPOSE: This study estimated the economic burden of breast cancer-related lymphedema (BCRL) among working-age women, the incidence of lymphedema, and associated risk factors. METHODS: We used claims data to study an incident cohort of breast cancer patients for the 2 years after the initiation of cancer treatment. A logistic regression model was used to ascertain factors associated with lymphedema. We compared the medical costs and rate of infections likely associated with lymphedema between a woman with BCRL and a matched control. We performed nonparametric bootstrapping to compare the unadjusted cost differences and estimated the adjusted cost differences in regression analysis. RESULTS: Approximately 10% of the 1,877 patients had claims indicating treatment of lymphedema. Predictors included treatment with full axillary node dissection (odds ratio [OR] = 6.3, P < .001) and chemotherapy (OR = 1.6, P = .01). A geographic variation was observed; women who resided in the West were more likely to have lymphedema claims than those in the Northeast (OR = 2.05, P = .01). The matched cohort analysis demonstrated that the BCRL group had significantly higher medical costs ($14,877 to $23,167) and was twice as likely to have lymphangitis or cellulitis (OR = 2.02, P = .009). Outpatient care, especially mental health services, diagnostic imaging, and visits with moderate or high complexity, accounted for the majority of the difference. CONCLUSION: Although the use of claims data may underestimate the true incidence of lymphedema, women with BCRL had a greater risk of infections and incurred higher medical costs. The substantial costs documented here suggest that further efforts should be made to elucidate reduction and prevention strategies for BCRL. PMID- 19289625 TI - Systematic review and meta-analysis of randomized and nonrandomized trials on safety and efficacy of video-assisted thoracic surgery lobectomy for early-stage non-small-cell lung cancer. AB - PURPOSE: The current randomized trials comparing video-assisted thoracic surgery (VATS) lobectomy with open lobectomy for patients with early-stage non-small-cell lung cancer (NSCLC) have been of small size. We performed the present meta analysis of the randomized and nonrandomized comparative studies in an attempt to assess the safety and efficacy of VATS lobectomy. METHODS: Electronic searches identified 21 eligible comparative studies (two randomized and 19 nonrandomized) for inclusion. Two reviewers independently appraised each study. Meta-analysis was performed by combining the results of reported incidence of morbidity and mortality, recurrence, and 5-year mortality rates. The relative risk (RR) was used as a summary statistic. RESULTS: There were no significant statistical differences between VATS and open lobectomy in terms of postoperative prolonged air leak (P = .71), arrhythmia (P = .86), pneumonia (P = .09), and mortality (P = .49). VATS did not demonstrate any significant difference in locoregional recurrence (P = .24), as compared with the open lobectomy arm, but the data suggested a reduced systemic recurrence rate (P = .03) and an improved 5-year mortality rate of VATS (P = .04). There was no evidence to suggest heterogeneity of trial results. Fourteen studies reported VATS to open lobectomy conversion rate ranging from 0% to 15.7% (median = 8.1%). CONCLUSION: Both randomized and nonrandomized trials suggest that VATS lobectomy is an appropriate procedure for selected patients with early-stage NSCLC when compared with open surgery. PMID- 19289626 TI - Pretreatment health behaviors predict survival among patients with head and neck squamous cell carcinoma. AB - PURPOSE: Our prior work has shown that the health behaviors of head and neck cancer patients are interrelated and are associated with quality of life; however, other than smoking, the relationship between health behaviors and survival is unclear. PATIENTS AND METHODS: A prospective cohort study was conducted to determine the relationship between five pretreatment health behaviors (smoking, alcohol, diet, physical activity, and sleep) and all-cause survival among 504 head and neck cancer patients. RESULTS: Smoking status was the strongest predictor of survival, with both current smokers (hazard ratio [HR] = 2.4; 95% CI, 1.3 to 4.4) and former smokers (HR = 2.0; 95% CI, 1.2 to 3.5) showing significant associations with poor survival. Problem drinking was associated with survival in the univariate analysis (HR = 1.4; 95% CI, 1.0 to 2.0) but lost significance when controlling for other factors. Low fruit intake was negatively associated with survival in the univariate analysis only (HR = 1.6; 95% CI, 1.1 to 2.1), whereas vegetable intake was not significant in either univariate or multivariate analyses. Although physical activity was associated with survival in the univariate analysis (HR = 0.95; 95% CI, 0.93 to 0.97), it was not significant in the multivariate model. Sleep was not significantly associated with survival in either univariate or multivariate analysis. Control variables that were also independently associated with survival in the multivariate analysis were age, education, tumor site, cancer stage, and surgical treatment. CONCLUSION: Variation in selected pretreatment health behaviors (eg, smoking, fruit intake, and physical activity) in this population is associated with variation in survival. PMID- 19289627 TI - Socioeconomic differences in patient survival are increasing for acute myeloid leukemia and multiple myeloma in sweden. AB - PURPOSE: An association between socioeconomic status (SES) and survival in acute myeloid leukemia (AML) and multiple myeloma (MM) has not been established in developed countries. We assessed the impact of SES on survival in two large population-based cohorts of AML and MM patients diagnosed in Sweden 1973 to 2005. PATIENTS AND METHODS: The relative risk of death (all cause and cause specific) in relation to SES was estimated using Cox's proportional hazards regression. We also conducted analyses stratified by calendar periods (1973 to 1979, 1980 to 1989, 1990 to 1999, and 2000 to 2005). RESULTS: We identified a total of 9,165 and 14,744 patients with AML and MM, respectively. Overall, higher white-collar workers had a lower mortality than other SES groups for both AML (P = .005) and MM (P < .005). In AML patients, a consistently higher overall mortality was observed in blue-collar workers compared with higher white-collar workers in the last three periods (hazard ratio [HR], 1.26; 95% CI, 1.05 to 1.51; HR, 1.23; 95% CI, 1.05 to 1.45; HR, 1.28; 95% CI, 1.04 to 1.57, respectively). In MM, no difference was observed in the first two calendar periods. However, in 1990 to 1999, self-employed (HR, 1.18; 95% CI, 1.02 to 1.37), blue-collar workers (HR, 1.18; 95% CI, 1.04 to 1.32), and retired (HR, 1.45; 95% CI, 1.16 to 1.80) had a higher mortality compared to higher white-collar workers. In 2000 to 2005, blue collar workers had a higher mortality (HR, 1.31; 95% CI, 1.07 to 1.60) compared with higher white-collar workers. CONCLUSION: SES was significantly associated with survival in both AML and MM. Most conspicuously, a lower mortality was observed among the highest SES group during more recent calendar periods. Differences in management, comorbidity, and lifestyle, are likely factors to explain these findings. PMID- 19289628 TI - Durable long-term remission with chemotherapy alone for stage II to IV laryngeal cancer. AB - PURPOSE: For patients with stage II to IV laryngeal cancer, radiation therapy (RT) either alone or with concurrent chemotherapy provides the highest rate of organ preservation but can be associated with functional impairment. Thus, we studied the use of induction chemotherapy with or without conservation laryngeal surgery (CLS). Our objectives were to study the sensitivity of laryngeal cancer to platinum-based chemotherapy alone and to highlight the efficacy of CLS in this setting. PATIENTS AND METHODS: Thirty-one previously untreated patients with laryngeal cancer (T2-4, N0-1, M0), who were resectable with CLS, were enrolled. Patients received three to four cycles of paclitaxel, ifosfamide, and cisplatin (TIP) chemotherapy, and response was assessed histologically. Patients with partial response (PR) proceeded to CLS. Patients achieving pathologic complete response (pCR) received an additional three cycles of TIP and no other treatment. RESULTS: Thirty patients were assessable for response. With TIP chemotherapy alone, 11 patients (37%) achieved pCR, 10 of whom (33%) remain alive with durable disease remission and no evidence of recurrence over a median follow-up time of 5 years. Nineteen patients (63%) treated with TIP alone achieved PR. The overall laryngeal preservation (LP) rate was 83%, and only five patients (16%) required postoperative RT. No patient required a gastrostomy tube or tracheotomy. CONCLUSION: Chemotherapy alone in selected patients with T2-4, N0-1 laryngeal cancer can provide durable disease remission at 5 years. For patients with PR, CLS provides a high rate of LP. This prospective study suggests that chemotherapy alone may cure selected patients with laryngeal cancer, warranting further prospective investigation. PMID- 19289629 TI - Genomic strategy for targeting therapy in castration-resistant prostate cancer. AB - PURPOSE: Despite treatments which lower circulating androgens, advanced prostate cancers often maintain androgen receptor (AR) signaling. The variable response to secondary hormonal manipulations in men with castrate-resistant prostate cancer (CRPC) creates a compelling need for strategies to individualize therapy based on the molecular features of each patient's tumor. METHODS: A transcription-based AR activity signature was developed from an androgen-sensitive prostate cancer cell (LNCaP) and tested on independent data sets of prostate cancer cell lines and human tumors to assess its precision and accuracy in detecting AR activity. The AR signature was applied to multiple sets of prostate specimens to determine how AR activity changes with hormone therapy and progression and oncogenic pathway analysis was used to identify biologic pathways correlating with AR activity. RESULTS: A robust AR signature accurately predicts AR activity in multiple prostate cancer cell lines, has minimal variation between replicate samples, and accurately reflects an individual's hormone status and intraprostatic dihydrotestosterone levels. The AR signature finds AR activity to be high in local, untreated prostate tumors and decreased in prostate tissue after neoadjuvant hormone therapy and in CRPC. Heterogeneity of AR activity exists along the spectrum of prostate cancer progression and decreasing predicted AR activity correlates with increasing predicted Src activity and sensitivity to dasatinib (Src-targeting kinase inhibitor). CONCLUSION: A transcription-based AR signature can detect AR activity within individual prostate cancer specimens and has the potential to help individualize and improve care for patients with CRPC. PMID- 19289630 TI - Phase III study of gefitinib compared with intravenous methotrexate for recurrent squamous cell carcinoma of the head and neck [corrected]. AB - PURPOSE: To compare survival in patients with recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN) treated with gefitinib 250 or 500 mg/day or standard methotrexate. PATIENTS AND METHODS: Four hundred eighty-six patients with recurrent SCCHN were randomly assigned to oral gefitinib 250 mg/day, gefitinib 500 mg/day, or methotrexate 40 mg/m(2) intravenously weekly. Primary end point was overall survival, secondary end points were objective response rate (ORR), safety, symptom improvement, and quality of life (QOL). Exploratory end points included association of efficacy with epidermal growth factor receptor gene copy number and other biomarkers. RESULTS: Neither gefitinib 250 nor 500 mg/day improved overall survival compared with methotrexate (hazard ratio [HR], 1.22; 95% CI, 0.95 to 1.57; P = .12; and HR, 1.12; 95% CI, 0.87 to 1.43; P = .39, respectively). In the gefitinib 250 mg/day, 500 mg/day, and methotrexate groups, respectively, median overall survival was 5.6, 6.0, and 6.7 months; ORRs (Response Evaluation Criteria in Solid Tumors) were 2.7%, 7.6% and 3.9%, with no statistically significant difference between either gefitinib arm and methotrexate. No unexpected adverse events were observed, except for tumor hemorrhage-type events with gefitinib (8.9%, gefitinib 250 mg/day; 11.4%, gefitinib 500 mg/day; 1.9%, methotrexate). QOL improvement rates (Functional Assessment of Cancer Therapy-Head & Neck total score) were 13.4%, 18.0%, and 6.0% for gefitinib 250 mg/day, 500 mg/day, and methotrexate, respectively. CONCLUSION: In patients with recurrent or metastatic SCCHN, while responses with gefitinib were seen, neither gefitinib 250 nor 500 mg/day improved overall survival compared with methotrexate. With the exception of tumor hemorrhage-type events with gefitinib, the adverse event profiles were generally consistent with those previously observed. PMID- 19289631 TI - Candidate genes on chromosome 9q33-34 involved in the progression of childhood ependymomas. AB - PURPOSE: The molecular pathogenesis of pediatric ependymoma remains unclear. Our study was designed to identify genetic changes implicated in ependymoma progression. PATIENTS AND METHODS: We characterized 59 ependymoma samples (33 at diagnosis and 26 at relapse) using array-comparative genomic hybridization (aCGH). Specific chromosomal imbalances were confirmed by fluorescent in situ hybridization, and candidate genes were assessed by real-time quantitative polymerase chain reaction (qPCR), immunohistochemistry, sequencing, and in vitro functional studies. RESULTS: aCGH analysis revealed a significant increase in genomic imbalances on relapse compared with diagnosis, such as gain of 9qter and 1q (54% v 21% and 12% v 0%, respectively) and loss of 6q (27% v 6%). Supervised tumor classification showed that gain of 9qter was associated with tumor recurrence, age older than 3 years, and posterior fossa location. Using a candidate-gene strategy, we found an overexpression of two potential oncogenes at the locus 9qter: Tenascin-C and Notch1. Moreover, Notch pathway analysis (qPCR) revealed overexpression of Notch ligands, receptors, and target genes (Hes-1, Hey2, and c-Myc), and downregulation of Notch repressor Fbxw7. We confirmed by immunohistochemistry the overexpression of Tenascin-C and Hes-1. We detected Notch1 missense mutations in 8.3% of the tumors (only in the posterior fossa location and in case of 9q33-34 gain). Furthermore, inhibition of Notch pathway with a gamma-secretase inhibitor impaired the growth of ependymoma stem cell cultures. CONCLUSION: The activation of the Notch pathway and Tenascin-C seem to be important events in ependymoma progression and may represent future targets for therapy. We report, to our knowledge for the first time, recurrent oncogenic mutations in pediatric posterior fossa ependymomas. PMID- 19289632 TI - A statewide collaborative initiative to improve the quality of care for patients with acute myocardial infarction and heart failure. AB - BACKGROUND: To enhance quality improvement, we created a unique statewide collaboration among 3 organizations: the Virginia Health Quality Center (Virginia's Medicare Quality Improvement Organization), the American College of Cardiology, and the American Heart Association. The goal was to improve discharge measures for acute myocardial infarction and heart failure. METHODS AND RESULTS: In 2004, 29 hospitals participated in the collaborative initiative. Using Medicare data submitted from 2004 through the second quarter of 2006, we analyzed adherence to individual discharge measures and all-or-none appropriate care measures for acute myocardial infarction, heart failure, and both. To control for differences in hospital characteristics, we were able to match 21 of the participating hospitals with 21 similar nonparticipating hospitals. In this paired analysis, the total appropriate care measure increased from 61% to 77% in participating hospitals compared with an increase from 51% to 60% in nonparticipating hospitals (P<0.0001). A generalized linear mixed model examining the full data set at the patient level failed to show a clear advantage among participating hospitals. Participating hospitals had higher baseline rates for most quality measures, suggesting a possible effect of a prior collaborative. Further analysis of only hospitals that participated in a prior collaborative showed that participants in the current collaborative initiative had higher rates of improvement for 7 of 10 quality measures and appropriate care measures for heart failure, acute myocardial infarction, or both (all P<0.05). CONCLUSIONS: We report a unique collaboration of a Medicare Quality Improvement Organization and 2 national organizations to address quality of care for acute myocardial infarction and heart failure. A composite measure of quality (the total appropriate care measure) improved more in the participating hospitals during the timeframe of the intervention, although the greater improvement in this and other measures in the participating hospitals appeared to be dependent on participation in a prior collaborative initiative. PMID- 19289633 TI - Plasma asymmetric dimethylarginine and incidence of cardiovascular disease and death in the community. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, induces endothelial dysfunction. Although elevated ADMA has been associated with an increased risk of cardiovascular disease events and death in referral samples, the prognostic significance of ADMA in the community has not been adequately evaluated. METHODS AND RESULTS: We related plasma ADMA, l arginine (Arg), and the ratio of Arg to ADMA to the incidence of cardiovascular disease (fatal or nonfatal myocardial infarction, coronary insufficiency, angina pectoris, stroke or transient ischemic attack, intermittent claudication, or heart failure) and death in 3320 Framingham Offspring Study participants (1769 women; mean age, 59 years). Over a follow-up period of 10.9 years, 281 individuals of 2956 free of cardiovascular disease at baseline developed incident cardiovascular disease, and 285 participants died. In multivariable models adjusting for established risk factors and other biomarkers (B-type natriuretic peptide, renin, homocysteine, urine albumin excretion, and C-reactive protein), ADMA and the ratio of Arg to ADMA were significantly associated with all-cause mortality (adjusted-hazard ratio [HR] per 1-SD increment, 1.21; 95% CI, 1.07 to 1.37; and HR per 1-SD increment, 0.80; 95% CI, 0.69 to 0.93, respectively), whereas Arg was not (HR per 1-SD increment, 0.89; 95% CI, 0.77 to 1.02). We noted effect modification by diabetes status; ADMA was associated with death in individuals without diabetes (adjusted HR per 1-SD increment, 1.30; 95% CI, 1.13 to 1.50) but not in individuals with diabetes (adjusted HR per 1-SD increment, 0.85; 95% CI, 0.62 to 1.16). ADMA, Arg, and the ratio of Arg to ADMA were not associated with cardiovascular disease incidence (P>0.10). CONCLUSIONS: In our large community-based sample, ADMA was significantly associated with all-cause mortality, particularly in nondiabetic individuals. PMID- 19289634 TI - Pericardial fat, intrathoracic fat, and measures of left ventricular structure and function: the Framingham Heart Study. AB - BACKGROUND: Pericardial fat has been implicated in the pathogenesis of obesity related cardiovascular disease. Whether the associations of pericardial fat and measures of cardiac structure and function are independent of the systemic effects of obesity and visceral adiposity has not been fully explored. METHODS AND RESULTS: Participants from the Framingham Heart Study (n=997; 54.4% women) underwent chest and abdominal computed tomography and cardiovascular magnetic resonance imaging between 2002 and 2005. Pericardial fat, intrathoracic fat, and visceral adipose tissue quantified from multidetector computed tomography, along with body mass index and waist circumference, were examined in relation to cardiovascular magnetic resonance measures of left ventricular (LV) mass, LV end diastolic volume, and left atrial dimension. In women, pericardial fat (r=0.20 to 0.35, P<0.001), intrathoracic fat (r=0.25 to 0.37, P<0.001), visceral adipose tissue (r=0.24 to 0.45, P<0.001), body mass index (r=0.36 to 0.53, P<0.001), and waist circumference (r=0.30 to 0.48, P<0.001) were directly correlated with LV mass, LV end-diastolic volume, and left atrial dimension. In men, pericardial fat (r=0.19 to 0.37, P<0.001), intrathoracic fat (r=0.17 to 0.31, P<0.001), visceral adipose tissue (r=0.19 to 0.36, P<0.001), body mass index (r=0.32 to 0.44, P<0.001), and waist circumference (r=0.34 to 0.44, P<0.001) were directly correlated with LV mass and left atrial dimension, but LV end-diastolic volume was not consistently associated with adiposity measures. Associations persisted after multivariable adjustment but not after additional adjustment for body weight and visceral adipose tissue, except for pericardial fat and left atrial dimension in men. CONCLUSIONS: Pericardial fat is correlated with cardiovascular magnetic resonance measures, but the association is not independent of or stronger than other ectopic fat stores or proxy measures of visceral adiposity. An important exception is left atrial dimension in men. These results suggest that the systemic effects of obesity on cardiac structure and function may outweigh the local pathogenic effects of pericardial fat. PMID- 19289635 TI - Detoxification and endothelialization of glutaraldehyde-fixed bovine pericardium with titanium coating: a new technology for cardiovascular tissue engineering. AB - BACKGROUND: Endothelial cell seeding of glutardialdehyde-fixed biological heart valves is hypothesized to improve biocompatibility and durability; however, the toxicity of glutardialdehyde prevents its use as a biological coating. Therefore, different detoxification strategies are applied, including surface coating with titanium, before in vitro endothelialization of glutaraldehyde-fixed bovine pericardium as the base material for prosthetic heart valves. METHODS AND RESULTS: Bovine pericardium was fixed with 0.25% glutardialdehyde. Detoxification was performed with citric acid, aldehyde dehydrogenase, and plasma deposition with titanium at low temperatures of 30 degrees C to 35 degrees C. Toxic glutaraldehyde ligands were quantified photometrically, and the vitality of seeded cells was tested to validate detoxification methods. Detoxification agents and titanium coating were applied before seeding with human endothelial cells. Endothelial cells were visualized by electron microscopic surface scanning. To evaluate cell adhesion, shear stress was applied by a flow of 5 L/min over 24 hours. Compared with untreated glutaraldehyde-fixed samples, treatment with the different agents reduced free aldehyde groups gradually (citric acid 5% < citric acid 10% < titanium < aldehyde dehydrogenase). A combination of citric acid 10%, aldehyde dehydrogenase, and titanium coating resulted in a reduction of free aldehyde ligands to 17.3+/-4.6% (P < or = 0.05) and demonstrated a vitality of seeded cells of 94+/-6.7% (P < or = 0.05). This procedure yielded a completely confluent layer of regular human endothelial cells (n=5). After application of shear stress for 24 hours on these endothelial layers, cell vitality was 81%. CONCLUSIONS: Titanium coating combined with chemical procedures yielded significant detoxification and complete endothelialization of conventional glutaraldehyde-fixed pericardium. This new technique might improve glutardialdehyde-fixed cardiovascular bioimplants for better biocompatibility and longer durability. PMID- 19289636 TI - Effects of different types of exercise training followed by detraining on endothelium-dependent dilation in patients with recent myocardial infarction. AB - BACKGROUND: In coronary artery disease, exercise training (ET) is associated with an improvement in endothelial function, but little is known about the relative effect of different types of training. The purpose of this study was to prospectively evaluate the effect of different types of ET on endothelial function in 209 patients after a first recent acute myocardial infarction. METHODS AND RESULTS: Endothelial function was evaluated before and after 4 weeks of different types of ET and after 1 month of detraining by measuring flow mediated dilation and von Willebrand factor levels at baseline and after ET. Patients were randomized into 4 groups: group 1, aerobic ET (n=52); group 2, resistance training (n=54); group 3, resistance plus aerobic training (n=53); and group 4, no training (n=50). At baseline, flow-mediated dilation was 4.5+/-2.6% in group 1, 4.01+/-1.6% in group 2, 4.4+/-4% in group 3, and 4.3+/-2.3% in group 4 (P=NS). After ET, flow-mediated dilation increased to 9.9+/-2.5% in group 1, 10.1+/-2.6% in group 2, and 10.8+/-3% in group 3 (P<0.01 versus baseline for all groups); it also increased in group 4 but to a much lesser extent (to 5.1+/-2.5%; P<0.01 versus trained groups). The von Willebrand factor level after ET decreased by 16% (P<0.01) similarly in groups 1, 2, and 3 but remained unchanged in group 4. Detraining returned flow-mediated dilation to baseline levels (P<0.01 versus posttraining). CONCLUSIONS: In patients with recent acute myocardial infarction, ET was associated with improved endothelial function independently of the type of training, but this effect disappeared after 1 month of detraining. PMID- 19289637 TI - Local inflammation and hypoxia abolish the protective anticontractile properties of perivascular fat in obese patients. AB - BACKGROUND: Inflammation in adipose tissue has been implicated in vascular dysfunction, but the local mechanisms by which this occurs are unknown. METHODS AND RESULTS: Small arteries with and without perivascular adipose tissue were taken from subcutaneous gluteal fat biopsy samples and studied with wire myography and immunohistochemistry. We established that healthy adipose tissue around human small arteries secretes factors that influence vasodilation by increasing nitric oxide bioavailability. However, in perivascular fat from obese subjects with metabolic syndrome (waist circumference 111+/-2.8 versus 91.1+/-3.5 cm in control subjects, P<0.001; insulin sensitivity 41+/-5.9% versus 121+/-18.6% in control subjects, P<0.001), the loss of this dilator effect was accompanied by an increase in adipocyte area (1786+/-346 versus 673+/-60 mum(2), P<0.01) and immunohistochemical evidence of inflammation (tumor necrosis factor receptor 1 12.4+/-1.1% versus 6.7+/-1%, P<0.001). Application of the cytokines tumor necrosis factor receptor-alpha and interleukin-6 to perivascular fat around healthy blood vessels reduced dilator activity, resulting in the obese phenotype. These effects could be reversed with free radical scavengers or cytokine antagonists. Similarly, induction of hypoxia stimulated inflammation and resulted in loss of anticontractile capacity, which could be rescued by catalase and superoxide dismutase or cytokine antagonists. Incubation with a soluble fragment of adiponectin type 1 receptor or inhibition of nitric oxide synthase blocked the vasodilator effect of healthy perivascular adipose tissue. CONCLUSIONS: We conclude that adipocytes secrete adiponectin and provide the first functional evidence that it is a physiological modulator of local vascular tone by increasing nitric oxide bioavailability. This capacity is lost in obesity by the development of adipocyte hypertrophy, leading to hypoxia, inflammation, and oxidative stress. PMID- 19289638 TI - Safety of short-term discontinuation of antiplatelet therapy in patients with drug-eluting stents. AB - BACKGROUND: Antiplatelet therapy is often discontinued in patients with drug eluting stents who are undergoing surgical procedures. However, discontinuation of antiplatelet therapy is an important risk factor for late stent thrombosis. Our objective was to examine the safety of short-term discontinuation of antiplatelet therapy. METHODS AND RESULTS: We systematically searched Medline for reported cases of late stent thrombosis and very late stent thrombosis published between January 2001 and July 2008. We restricted our search to Academic Research Consortium-defined definite cases. We identified 161 cases of late stent thrombosis or very late stent thrombosis from 84 articles (79 from case reports, 61 from registries, and 21 from randomized clinical trials). Patients had a mean age of 58.4+/-13.4 years, and 88% were male. A total of 19 cases occurred in patients who were receiving dual antiplatelet therapy at the time of the event. If patients stopped both antiplatelet agents simultaneously, the median time to event was 7 days. If patients had previously stopped a thienopyridine with no ill effect and subsequently stopped acetylsalicylic acid, the median time to event was also 7 days from the time of acetylsalicylic acid cessation. If the thienopyridine was stopped but acetylsalicylic acid was maintained, the median time to event was 122 days. Among the 48 patients who stopped both agents, 36 cases (75%) occurred within 10 days. Among the 94 patients who discontinued a thienopyridine but continued acetylsalicylic acid, only 6 cases (6%) occurred within 10 days. CONCLUSIONS: If acetylsalicylic acid therapy is maintained, short term discontinuation of a thienopyridine may be relatively safe in patients with drug-eluting stents. PMID- 19289639 TI - Molecular architecture of the human sinus node: insights into the function of the cardiac pacemaker. AB - BACKGROUND: Although we know much about the molecular makeup of the sinus node (SN) in small mammals, little is known about it in humans. The aims of the present study were to investigate the expression of ion channels in the human SN and to use the data to predict electrical activity. METHODS AND RESULTS: Quantitative polymerase chain reaction, in situ hybridization, and immunofluorescence were used to analyze 6 human tissue samples. Messenger RNA (mRNA) for 120 ion channels (and some related proteins) was measured in the SN, a novel paranodal area, and the right atrium (RA). The results showed, for example, that in the SN compared with the RA, there was a lower expression of Na(v)1.5, K(v)4.3, K(v)1.5, ERG, K(ir)2.1, K(ir)6.2, RyR2, SERCA2a, Cx40, and Cx43 mRNAs but a higher expression of Ca(v)1.3, Ca(v)3.1, HCN1, and HCN4 mRNAs. The expression pattern of many ion channels in the paranodal area was intermediate between that of the SN and RA; however, compared with the SN and RA, the paranodal area showed greater expression of K(v)4.2, K(ir)6.1, TASK1, SK2, and MiRP2. Expression of ion channel proteins was in agreement with expression of the corresponding mRNAs. The levels of mRNA in the SN, as a percentage of those in the RA, were used to estimate conductances of key ionic currents as a percentage of those in a mathematical model of human atrial action potential. The resulting SN model successfully produced pacemaking. CONCLUSIONS: Ion channels show a complex and heterogeneous pattern of expression in the SN, paranodal area, and RA in humans, and the expression pattern is appropriate to explain pacemaking. PMID- 19289640 TI - Randomized trial of warfarin, aspirin, and clopidogrel in patients with chronic heart failure: the Warfarin and Antiplatelet Therapy in Chronic Heart Failure (WATCH) trial. AB - BACKGROUND: Chronic heart failure remains a major cause of mortality and morbidity. The role of antithrombotic therapy in patients with chronic heart failure has long been debated. The objective of this study was to determine the optimal antithrombotic agent for heart failure patients with reduced ejection fractions who are in sinus rhythm. METHODS AND RESULTS: This prospective, randomized clinical trial of open-label warfarin (target international normalized ratio of 2.5 to 3.0) and double-blind treatment with either aspirin (162 mg once daily) or clopidogrel (75 mg once daily) had a 30-month enrollment period and a minimum of 12 months of treatment. We enrolled 1587 men and women >/=18 years of age with symptomatic heart failure for at least 3 months who were in sinus rhythm and had left ventricular ejection fraction of 50% (P<0.01) and for the beta-subunit minK by approximately 42% (P<0.05). SAN transcript expression for the rapid delayed-rectifier (I(Kr)) alpha subunit ERG, the slow delayed-rectifier (I(Ks)) alpha-subunit KvLQT1, the beta subunit MiRP1, the L-type (I(CaL)) and T-type (I(CaT)) Ca2+-current subunits Cav1.2 and Cav3.1, and the gap-junction subunit connexin 43 (were unaffected by atrial tachypacing. Atrial tachypacing reduced densities of the HCN-related funny current (I(f)) and I(Ks) by approximately 48% (P<0.001) and approximately 34% (P<0.01), respectively, with no change in voltage dependence or kinetics. I(Kr), I(CaL), and I(CaT) were unaffected. SAN cells lacked Ba2+-sensitive inward rectifier currents, irrespective of AT. SAN action potential simulations that incorporated AT-induced alterations in I(f) accounted for slowing of periodicity, with no additional contribution from changes in I(Ks). CONCLUSIONS: AT downregulates SAN HCN2/4 and minK subunit expression, along with the corresponding currents I(f) and I(Ks). Tachycardia-induced remodeling of SAN ion channel expression, particularly for the "pacemaker" subunit I(f), may contribute to the clinically significant association between SAN dysfunction and supraventricular tachyarrhythmias. PMID- 19289642 TI - Resveratrol prevents the prohypertrophic effects of oxidative stress on LKB1. AB - BACKGROUND: Master regulators of protein synthesis such as mammalian target of rapamycin (mTOR) and p70S6 kinase contribute to left ventricular hypertrophy. These prohypertrophic pathways are modulated by a number of kinase cascades, including the hierarchical LKB1/AMP-activated protein kinase (AMPK) energy sensing pathway. Because oxidative stress inhibits the LKB1/AMPK signaling axis to promote abnormal cell growth in cancer cells, we investigated whether oxidative stress associated with hypertension also results in the inhibition of this kinase circuit to contribute to left ventricular hypertrophy. METHODS AND RESULTS: In the spontaneously hypertensive rat, a well-established genetic model of hypertension and subsequent cardiac hypertrophy, the development of left ventricular hypertrophy is associated with an increase in the electrophilic lipid peroxidation byproduct 4-hydroxy-2-nonenal (HNE). Using isolated cardiomyocytes, we show that elevated levels of HNE result in the formation of HNE-LKB1 adducts that inhibit LKB1 and subsequent AMPK activity. Consistent with inhibition of the LKB1/AMPK signaling pathway, the mTOR/p70S6 kinase system is activated, which is permissive for cardiac myocyte cell growth. Treatment of cardiomyocytes with resveratrol prevents HNE modification of the LKB1/AMPK signaling axis and blunts the prohypertrophic p70S6 kinase response. Furthermore, administration of resveratrol to spontaneously hypertensive rats results in increased AMPK phosphorylation and activity and reduced left ventricular hypertrophy. CONCLUSIONS: Our data identify a molecular mechanism in the cardiomyocyte involving the oxidative stress-derived lipid peroxidation byproduct HNE and the LKB1/AMPK signaling pathway that contributes to the development of left ventricular hypertrophy. We also suggest that resveratrol may be a potential therapy for patients at risk for developing pathological cardiac hypertrophy by preventing this prohypertrophic process. PMID- 19289643 TI - Effect of sulfaphenazole on tissue plasminogen activator release in normotensive subjects and hypertensive patients. AB - BACKGROUND: A nitric oxide-independent response, possibly mediated by hyperpolarization, regulates vascular tone, acting as a compensatory mechanism in the presence of impaired nitric oxide availability. Cytochrome P450 2C9 (CYP 2C9) is a source of endothelium-derived hyperpolarizing factors and modulates tissue type plasminogen activator (tPA) release from endothelial cells; however, no effect of hyperpolarization on fibrinolysis has been documented in humans. We aimed to assess the effect of sulfaphenazole, a specific CYP 2C9 inhibitor, on tPA release in normotensive subjects and patients with essential hypertension. METHODS AND RESULTS: tPA release was measured in the forearm microcirculation of 56 normotensivesubjects and 57 patients with essential hypertension after bradykinin (0.015 microg x 100 mL(-1) x min(-1)) and acetylcholine (1.5 microg x 100 mL(-1) x min(-1)) infusions, with or without sulfaphenazole (0.03 microg x 100 mL(-1) x min(-1)). Bradykinin and acetylcholine infusions were repeated with N(G)-monomethyl-l-arginine (L-NMMA; 100 microg x 100 mL(-1) x min(-1)) and/or sulfaphenazole. tPA release by bradykinin and acetylcholine was higher in normotensive subjects than in patients with essential hypertension (P<0.01). Sulfaphenazole (P<0.01) blunted bradykinin-induced but not acetylcholine-induced tPA release in both groups. In normotensive subjects, L-NMMA infusion reduced tPA release (P<0.01). When L-NMMA was coinfused with sulfaphenazole, tPA release induced by bradykinin, but not by acetylcholine, was further reduced (P<0.01). In patients with essential hypertension, tPA release by both agonists was unaffected by L-NMMA, but only bradykinin-induced tPA release was blunted by sulfaphenazole, alone or with L-NMMA (P<001). CONCLUSIONS: Sulfaphenazole inhibits bradykinin induced tPA release, which suggests a modulatory role of CYP 2C9-derived endothelium-derived hyperpolarizing factors in tPA release in humans. In patients with essential hypertension, tPA release depends exclusively on endothelium derived hyperpolarizing factor, which is an ineffective compensatory mechanism in the presence of impaired nitric oxide availability. PMID- 19289644 TI - Images in cardiovascular medicine. Aneurysm rupture after stent grafting: value of dynamic imaging. PMID- 19289645 TI - Letter by Hartog et al. regarding article, "Advanced glycation end products accumulate in vascular smooth muscle and modify vascular but not ventricular properties in elderly hypertensive canines". PMID- 19289646 TI - Expression of concern. NCX-4016 (NO-aspirin) inhibits lipopolysaccharide-induced tissue factor expression in vivo: role of nitric oxide. PMID- 19289648 TI - Cocoa and cardiovascular health. AB - Epidemiological data demonstrate that regular dietary intake of plant-derived foods and beverages reduces the risk of coronary heart disease and stroke. Among many ingredients, cocoa might be an important mediator. Indeed, recent research demonstrates a beneficial effect of cocoa on blood pressure, insulin resistance, and vascular and platelet function. Although still debated, a range of potential mechanisms through which cocoa might exert its benefits on cardiovascular health have been proposed, including activation of nitric oxide and antioxidant and antiinflammatory effects. This review summarizes the available data on the cardiovascular effects of cocoa, outlines potential mechanisms involved in the response to cocoa, and highlights the potential clinical implications associated with its consumption. PMID- 19289649 TI - Qualitative and mixed methods provide unique contributions to outcomes research. PMID- 19289650 TI - National Heart, Lung, and Blood Institute: transitions. PMID- 19289651 TI - Angiotensin I is largely converted to angiotensin (1-7) and angiotensin (2-10) by isolated rat glomeruli. AB - Intraglomerular renin-angiotensin system enzyme activities have been examined previously using glomerular lysates and immune-based assays. However, preparation of glomerular extracts compromises the integrity of their anatomic architecture. In addition, antibody-based assays focus on angiotensin (Ang) II detection, ignoring the generation of other Ang I-derived metabolites, some of which may cross-react with Ang II. Therefore, our aim was to examine the metabolism of Ang I in freshly isolated intact glomeruli using matrix-assisted laser desorption ionization time of flight mass spectrometry as an analytic method. Glomeruli from male Sprague-Dawley rats were isolated by sieving and incubated in Krebs buffer in the presence of 1 micromol/L of Ang I for 15 to 90 minutes, with or without various peptidase inhibitors. Peptide sequences were confirmed by matrix-assisted laser desorption ionization time of flight tandem mass spectrometry or linear trap-quadrupole mass spectrometry. Peaks were quantified using customized valine (13)C(.15)N-labeled peptides as standards. The most prominent peaks resulting from Ang I cleavage were 899 and 1181 m/z, corresponding with Ang (1-7) and Ang (2-10), respectively. Smaller peaks for Ang II, Ang (1-9), and Ang (3-10) also were detected. The disappearance of Ang I was significantly reduced during inhibition of aminopeptidase A or neprilysin. In contrast, captopril did not alter Ang I degradation. Furthermore, during simultaneous inhibition of aminopeptidase A and neprilysin, the disappearance of Ang I was markedly attenuated compared with all of the other conditions. These results suggest that there is prominent intraglomerular conversion of Ang I to Ang (2-10) and Ang (1 7), mediated by aminopeptidase A and neprilysin, respectively. Formation of these alternative Ang peptides may be critical to counterbalance the local actions of Ang II. Enhancement of these enzymatic activities may constitute potential therapeutic targets for Ang II-mediated glomerular diseases. PMID- 19289652 TI - The natriuretic peptide uroguanylin elicits physiologic actions through 2 distinct topoisomers. AB - The peptide uroguanylin regulates electrolyte transport in the intestine and kidney. Human uroguanylin has 2 conformations that can be stably isolated because of their slow interconversion rate. The A isomer potently activates the guanylate cyclase C receptor found primarily in the intestine. The B isomer, by contrast, is a very weak agonist of this receptor, leading to a widely held assumption that it is physiologically irrelevant. We show here, however, that human uroguanylin B has potent natriuretic activity in the kidney. Interestingly, uroguanylin A and B both induce saluretic responses, but the activity profiles for the 2 peptides differ markedly. The uroguanylin B dose-response curve is sigmoidal with a threshold dose of approximately 10 nmol/kg of body weight, whereas uroguanylin A has a comparable threshold but a bell-shaped dose-response curve. In addition, our study indicates a unique interplay between the A and B isoforms, such that the A form at high concentrations antagonizes the natriuretic action of the B form. These data show that the kidney contains a uroguanylin receptor of which the pharmacological profile does not match that of the well-defined intestinal uroguanylin receptor (guanylate cyclase C), an observation consistent with previous studies showing that the kidney of the guanylate cyclase C knockout mouse remains responsive to uroguanylin. The results presented here also support the unconventional notion that distinct conformations of a single endocrine peptide can elicit different responses in different tissues. PMID- 19289653 TI - Defects in cutaneous angiotensin-converting enzyme 2 and angiotensin-(1-7) production in postural tachycardia syndrome. AB - Postural tachycardia syndrome (POTS) is associated with increased plasma angiotensin II (Ang II). Ang II administered in the presence of NO synthase inhibition with nitro-L-arginine (NLA) and Ang II type 1 receptor blockade with losartan produces vasodilation during local heating in controls. We tested whether this angiotensin-mediated vasodilation occurs in POTS and whether it is related to angiotensin-converting enzyme 2 (ACE2) and Ang-(1-7). We used local cutaneous heating to 42 degrees C and laser Doppler Flowmetry to assess NO dependent conductance at 4 calf sites in 12 low-flow POTS and in 12 control subjects 17.6 to 25.5 years of age. We perfused Ringer's solution through intradermal microdialysis catheters and performed local heating. We perfused one catheter with NLA (10 mmol/L)+losartan (2 microg/L) and repeated heating, and NLA+losartan+Ang II (10 micromol/L), repeating heating a third time. A second catheter received NLA+losartan+Ang II, heated, perfused NLA+losartan+Ang II+DX600 (1 mmol/L; a selective ACE2 inhibitor), and reheated. A third catheter received NLA+losartan+Ang II, heated, perfused NLA+losartan+Ang II+Ang-(1-7) (100 micromol/L), and reheated. The fourth catheter received Ang-(1-7) then reheated a second time only. Angiotensin-mediated vasodilation was present in control but not POTS. Ang-mediated dilation was eliminated by DX600, indicating an ACE2 related effect. Ang-mediated vasodilation was restored in POTS by Ang-(1-7). When administered alone during locally mediated heating, Ang-(1-7) improved the NO dependent local heating response. ACE2 effects are blunted in low-flow POTS and restored by the ACE2 product Ang-(1-7). Data imply impaired catabolism of Ang II through the ACE2 pathway. Vasoconstriction in POTS may result from a reduction in Ang-(1-7) and an increase in Ang II. PMID- 19289654 TI - Inhibition of tumor necrosis factor-alpha-induced interleukin-6 expression by telmisartan through cross-talk of peroxisome proliferator-activated receptor gamma with nuclear factor kappaB and CCAAT/enhancer-binding protein-beta. AB - Telmisartan, an angiotensin II type 1 receptor antagonist, was reported to be a partial agonist of peroxisome proliferator-activated receptor-gamma. Although peroxisome proliferator-activated receptor-gamma activators have been shown to have an anti-inflammatory effect, such as inhibition of cytokine production, it has not been determined whether telmisartan has such effects. We examined whether telmisartan inhibits expression of interleukin-6 (IL-6), a proinflammatory cytokine, in vascular smooth muscle cells. Telmisartan, but not valsartan, attenuated IL-6 mRNA expression induced by tumor necrosis factor-alpha (TNF alpha). Telmisartan decreased TNF-alpha-induced IL-6 mRNA and protein expression in a dose-dependent manner. Because suppression of IL-6 mRNA expression was prevented by pretreatment with GW9662, a specific peroxisome proliferator activated receptor-gamma antagonist, peroxisome proliferator-activated receptor gamma may be involved in the process. Telmisartan suppressed IL-6 gene promoter activity induced by TNF-alpha. Deletion analysis suggested that the DNA segment between -150 bp and -27 bp of the IL-6 gene promoter that contains nuclear factor kappaB and CCAAT/enhancer-binding protein-beta sites was responsible for telmisartan suppression. Telmisartan attenuated TNF-alpha-induced nuclear factor kappaB- and CCAAT/enhancer-binding protein-beta-dependent gene transcription and DNA binding. Telmisartan also attenuated serum IL-6 level in TNF-alpha-infused mice and IL-6 production from rat aorta stimulated with TNF-alpha ex vivo. These data suggest that telmisartan may attenuate inflammatory process induced by TNF alpha in addition to the blockade of angiotensin II type 1 receptor. Because both TNF-alpha and angiotensin II play important roles in atherogenesis through enhancement of vascular inflammation, telmisartan may be beneficial for treatment of not only hypertension but also vascular inflammatory change. PMID- 19289657 TI - Hybrid cardiac surgery: a resident's perspective. PMID- 19289658 TI - Multi-institutional experience using human acellular dermal matrix for ventral hernia repair in a compromised surgical field. AB - BACKGROUND: A complex ventral hernia repair (CVHR) involves a compromised surgical field where gastrointestinal, biliary, and genitourinary procedures are performed. Complex ventral hernia is a significant problem in trauma, emergency, and elective general surgery in which prosthetic material is contraindicated. In this clinical scenario, primary fascia closure carries a 50% risk of developing a hernia. The other option is a planned ventral hernia with delayed repair. HYPOTHESIS: Human acellular dermal matrix is a suitable implant for CVHR in a compromised surgical field. DESIGN: Multi-institutional, 5-year retrospective review. SETTING: Four academic medical centers. PATIENTS AND METHODS: Each center obtained institutional review board approval. Patients included in the review had undergone CVHR with human acellular dermal matrix. Data collected included age, body mass index (calculated as weight in kilograms divided by height in meters squared), comorbidities, size of fascial defect, wound classification, hospital length of stay, length of follow-up, and mortality. Primary outcomes were surgical site infection, fistula recurrence, and hernia recurrence. Both chi2 and logistic regression analyses were performed. RESULTS: Two hundred forty patients met the study criteria. Their mean (SD) age was 52.2 (15.0) years, and 132 (55.0%) were men. The most common comorbidity was hypertension (115 patients [47.9%]), and the mean defect size was 201 cm2. The mean hospital length of stay was 17.2 days, and the mean follow-up was 317 days. The overall mortality was 2.9%. The hernia recurrence rate was 17.1% (41 patients). Repair of a fistula or stoma was associated with hernia recurrence (P = .03) and with fistula recurrence (P < .001). Logistic regression analysis demonstrated surgical site infection and body mass index of greater than 30 to be independent risks of hernia recurrence. CONCLUSIONS: Human acellular dermal matrix is a suitable alternative for CVHR in a compromised surgical field. The hernia recurrence rate with human acellular dermal matrix in a compromised surgical field is less than that seen with primary repair, offering additional and improved surgical options for CVHR in this group of patients. Stoma or fistula takedown at the time of CVHR continues to be associated with significant complications. PMID- 19289659 TI - Influence of obesity on cancer-related outcomes after pancreatectomy to treat pancreatic adenocarcinoma. AB - OBJECTIVE: To examine the influence of obesity, as measured by body mass index (BMI) (calculated as weight in kilograms divided by height in meters squared), on clinicopathologic factors and survival after pancreatectomy to treat adenocarcinoma. DESIGN: Retrospective review and statistical analysis using prospectively collected data. SETTING: Referral center with a dedicated multidisciplinary pancreas cancer program. PATIENTS: Two hundred eighty-five consecutive patients with data available for BMI calculation who underwent potentially curative pancreas resection to treat adenocarcinoma from January 1, 1999, to October 31, 2006. MAIN OUTCOME MEASURE: Influence of BMI and other known prognostic variables on the incidence of lymph node metastasis and disease-free and overall survival. RESULTS: We identified a subset of obese patients (BMI >35) who were at 12-fold risk of lymph node metastasis compared with nonobese patients (BMI < or =35). The estimated disease-free and overall survival rates were decreased in the obese patients, and the risk of cancer recurrence and death after pancreatectomy was nearly twice that in nonobese patients. CONCLUSIONS: Obese patients with a BMI of more than 35 are more likely to have node-positive pancreatic cancer and decreased survival after surgical resection. Data suggest that the negative influence of BMI of more than 35 on cancer-related end points is unrelated to the potential complexity of performing major oncologic surgery in obese patients. PMID- 19289660 TI - Combined aortic debranching and thoracic endovascular aneurysm repair (TEVAR) effective but at a cost. AB - OBJECTIVES: To compare hybrid repair (HR) (aortic debranching and TEVAR) with conventional open thoracoabdominal and aortic arch repairs (OR), including a cost analysis. DESIGN: Retrospective cohort. SETTING: University hospital. PATIENTS: Thirty patients with thoracoabdominal aneurysms were evaluated between November 1, 2005, and December 31, 2006. INTERVENTIONS: There were 18 HRs and 12 ORs. Aortic abnormalities included the arch, visceral aorta, and arch/visceral aorta combined. Aortic debranching with TEVAR (HR) was performed at a single setting. Dacron grafts were used for OR, and branch vessels were bypassed. Hospital costs and reimbursements were obtained from the finance department. MAIN OUTCOME MEASURES: Perioperative morbidity, mortality, and cost. RESULTS: Patients were significantly older in the HR group (mean [SD], 72 [8.9] vs 58 [17.4] years, P = .2). The HR group had significantly less blood loss (mean [SD], 1.7 [2.3] vs 4.8 [3.1] L, P = .004), transfusions (5.1 [5.9] vs 14.7 [7.8] units, P = .001), renal failure (0% vs 42.0%, P = .002), and pulmonary morbidity (17% vs 67%, P < .001); shorter intensive care unit stays (5.2 [4.8] vs 16.4 [12.9] days, P = .005); and shorter hospital length of stay (mean [SD], 11.6 [6.2] vs 20.8 [10.8] days, P = .01). There were no differences in mortality or spinal cord ischemia. There was no difference in mean direct hospital costs (HR: $59,435.70 vs OR: $49,341; P = .35). However, the mean cost margin per case was -34% for HR and +6.2% for OR (P = .04). CONCLUSIONS: Improved clinical outcomes are seen after HR despite treatment of an older, sicker patient population. However, HR ultimately comes at a significant cost to the hospital, with a 34% loss in revenue per case. PMID- 19289661 TI - Laparoscopic incisional hernia repair after solid-organ transplantation. AB - HYPOTHESIS: Laparoscopic incisional hernia repair (LIHR) is efficacious in transplant recipients. DESIGN: Retrospective review. SETTING: University hospital. PATIENTS: Thirty-one transplant recipients who underwent LIHR between July 9, 2004, and October 27, 2005. MAIN OUTCOME MEASURES: Operative complications and incisional hernia recurrence. RESULTS: The mean (SD) mesh size required for LIHR was 611 (307) cm2. Median (range) hospital stay was 4 (1-28) days, with follow-up of 589 (22-953) days. Eighteen patients developed a postoperative complication, most frequently seroma formation, which occurred in 13 patients (72%). The mesh size required for LIHR was significantly larger in patients with a postoperative complication (n = 18; 706 [319] cm2 vs n = 13; 480 [244] cm2; P = .04). Seroma formation was not associated with previous open hernia repair, diabetes mellitus, or corticosteroid use. No statistically significant relationship was noted between the transplanted organ and seroma development. There were no post-LIHR wound infections. In 7 patients (23%), hernia recurred. CONCLUSIONS: Laparoscopic incisional hernia repair in solid organ transplant recipients is associated with a high rate of seroma formation but minimal long-term morbidity. The recurrence rate after LIHR is equivalent to that after open hernia repair. These results suggest that LIHR is a safe and effective alternative to open repair in this patient population. PMID- 19289662 TI - Laparoscopic colorectal resection for bowel endometriosis: feasibility, complications, and clinical outcome. AB - OBJECTIVE: To evaluate the short- and long-term outcomes of laparoscopic colorectal resection for endometriosis. DESIGN AND PATIENTS: This study included 357 consecutive patients who underwent colorectal resection. We evaluated intraoperative and postoperative complications, symptom outcomes, and long-term follow-up. MAIN OUTCOME MEASURE: Three hundred forty-three patients (96.1%) underwent laparoscopic colorectal resection, and radical endometriosis ablation was in 334 patients (93.6%). RESULTS: Fourteen (3.9%) required laparoconversion. Median operating time was 300 (range, 85-720) minutes, with a median estimated blood loss of 250 (range, 50-550) mL. Radical endometriosis ablation was achieved in 334 patients (93.6%). Median ileus was 4 (range, 1-8) days, with a median postoperative hospitalization of 8 (range, 3-36) days. Early and late complications were observed in 44 patients (12.3%) and, in 35 of these (79.5%), surgical management was necessary. Median follow-up after colorectal resection was 19.6 (range, 6-48) months. The median preoperative and postoperative dyspareunia scores were 8 (range, 4-10) and 3 (range, 0-10), respectively (P < .04), and the median preoperative and postoperative gastrointestinal tract symptom scores were 7 (range, 2-10) and 2 (range, 0-10), respectively (P < .05). During follow-up, 24 of 286 recurrences (8.4%) were registered. Patients who previously underwent surgery for endometriosis showed a higher risk of recurrence compared with patients undergoing primary surgery (13.2% vs 3.4%; P < .048). CONCLUSIONS: Laparoscopic colorectal resection for severe endometriosis is feasible and markedly improved endometriosis-related symptoms. Despite the risk of major postoperative complications, the procedure shows good results in terms of recurrence rate and could be adopted as the primary approach for patients with symptomatic colorectal infiltrating endometriosis. PMID- 19289663 TI - Pretransplantation patient characteristics and survival following combined heart and kidney transplantation: an analysis of the United Network for Organ Sharing Database. AB - HYPOTHESIS: Pretransplantation patient characteristics determine survival following combined heart and kidney transplantation (HKT). DESIGN: Time-to-event analysis. SETTING: Academic research. PATIENTS: The United Network for Organ Sharing provided deidentified patient-level data. Analysis included 19,373 heart transplant recipients from January 1, 1995, to December 31, 2005. MAIN OUTCOME MEASURES: Multivariate Cox proportional hazards regression analysis was performed to identify pretransplantation recipient characteristics associated with improved long-term survival following HKT. Kaplan-Meier survival functions and Cox proportional hazards regression were used for time-to-event analysis. Using the relative risks calculated in regression analysis, weights were assigned for each risk factor, allowing for the construction of a risk score. RESULTS: Among heart transplant recipients, 264 (1.4%) underwent HKT. Factors associated with diminished survival included peripheral vascular disease, recipient age older than 65 years, nonischemic etiology of heart failure, dialysis dependence at the time of transplantation, and bridge to transplantation using a ventricular assist device. After stratification by risk score, 1-year survival was 93.2% and 61.9% in the lowest- and highest-risk HKT groups, respectively. Further stratification by estimated glomerular filtration rate (eGFR) was performed based on a previous study showing decreased survival of patients undergoing orthotopic heart transplantation with a preoperative eGFR of less than 33 mL/min. Low-risk patients with an eGFR of less than 33 mL/min undergoing HKT constituted the only group that had significantly better survival compared with isolated patients undergoing orthotopic heart transplantation with eGFRs and risk scores in the same range (P = .006). CONCLUSIONS: When patients were stratified by risk score and by diminished eGFR (<33 mL/min), low-risk HKT recipients with a diminished eGFR had improved survival following HKT over isolated heart transplant recipients. Only low-risk patients with combined kidney failure (eGFR, <33 mL/min) and heart failure seem to gain a survival benefit from HKT. PMID- 19289664 TI - Trends in management and prognosis for esophageal cancer surgery: twenty-five years of experience at a single institution. AB - OBJECTIVE: To investigate trends in results of esophagectomies to treat esophageal cancer at a single high-volume institution during the past 25 years. DESIGN AND SETTING: Retrospective cohort study in a university tertiary referral center. PATIENTS AND METHODS: Patients with cancer of the thoracic esophagus or esophagogastric junction seen from 1980 through 2004 were included (N = 3493). Three time periods were defined: 1980-1987, 1988-1995, and 1996-2004. MAIN OUTCOME MEASURES: Clinical presentation, tumor characteristics, and morbidity, mortality, and survival rates among patients with esophageal cancer undergoing esophagectomy. RESULTS: The ratio of squamous cell carcinoma to adenocarcinoma decreased from 3.3 to 1.7 (P <.001) during the study period, in parallel with an increase in the number of patients with tumors in the lower esophagus/esophagogastric junction. An increasing proportion of patients who underwent resection received neoadjuvant treatment (chemotherapy/chemoradiotherapy), and 1978 patients underwent esophagectomy. The R0 resection rate increased from 74.5% to 90.1% (P <.001). In addition, an increasing proportion of patients had early-stage tumor in the resected specimen. In-hospital postoperative mortality decreased from 8.2% to 2.6% (P <.001), and the 5-year survival rate significantly improved from 18.8% to 42.3% (P <.001) for all patients who underwent resection. Pathological tumor stage, completeness of the resection, time period, sex, tumor histological type, and tumor location influenced the prognosis of patients with esophageal cancer undergoing esophagectomy. CONCLUSIONS: A change in location and histological type of esophageal cancer has occurred during the past 25 years. Earlier diagnosis, a multidisciplinary approach, and refinements in surgical technique and perioperative care have led to a significant reduction in postoperative mortality rate and improved long-term survival among patients with cancer of the thoracic esophagus or esophagogastric junction. PMID- 19289665 TI - Effect of laparoscopy on the indications for adrenalectomy. AB - BACKGROUND: Laparoscopic adrenalectomy is now the criterion standard for removal of most benign adrenal lesions and may be used for malignant lesions as well. At the same time, improved imaging has led to an increase in the number of incidentally detected adrenal masses. The aim of this study was to determine whether the introduction of laparoscopy has changed the indications for adrenalectomy. DESIGN: Retrospective cohort study of patients operated on for primary adrenal disease between September 1, 1987, and August 17, 2007. SETTING: Academic hospital. PATIENTS: Sixty-six patients treated before (group 1) and 203 treated after (group 2) introduction of laparoscopic adrenalectomy. MAIN OUTCOME MEASURES: Patient characteristics, comorbidity, tumor size, indication, and time between diagnosis and surgery. RESULTS: Group 2 had more patients in American Society of Anesthesiologists class III with gastrointestinal and metabolic endocrine comorbidities. Tumor size did not change, and, despite an increase in the number of adrenalectomies, indications for surgery remained consistent over time. CONCLUSION: Despite an increased volume of procedures, the introduction of laparoscopic adrenalectomy in our hospital did not change the indications for surgical intervention. PMID- 19289666 TI - Impact of radiologic intervention on mortality in necrotizing pancreatitis: the role of organ failure. AB - BACKGROUND: Our group previously reported that organ failure and mortality in necrotizing pancreatitis (NP) are not different between patients with infected and sterile necrosis. Since that report, management of this disease has evolved to include image-guided percutaneous catheter drainage (PCD) to improve morbidity and mortality. We evaluated the effect of PCD on mortality in NP. DESIGN: Retrospective analysis. SETTING: Tertiary care referral center. PATIENTS: A total of 689 consecutive patients treated for acute pancreatitis between 2001 and 2005, of whom 64 (9.3%) had pancreatic necrosis documented on contrast-enhanced computed tomography. MAIN OUTCOME MEASURES: Mortality and organ failure. RESULTS: In the 64 patients with documented NP, overall mortality was 16%. Thirty-six patients (56%) had organ failure according to the Atlanta classification. Compared with patients with sterile necrosis, those with infected necrosis did not have an increased prevalence of organ failure or increased need for intubation, pressors, or dialysis but had an increased mortality. Mortality in patients treated conservatively was 1 of 29 (3%); in those with PCD alone, 6 of 11 (55%); in those with PCD and surgery, 2 of 17 (12%); and in those with surgery alone, 1 of 7 (14%). All patients treated with PCD alone had organ failure, whereas 10 (59%) of those with PCD and surgery had organ failure. CONCLUSION: The use of PCD did not improve the mortality of NP among patients with organ failure. PMID- 19289667 TI - Long-term outcomes of stapled hemorrhoidopexy vs conventional hemorrhoidectomy: a meta-analysis of randomized controlled trials. AB - OBJECTIVES: To assess the long-term results of stapled hemorrhoidopexy (SH) compared with conventional hemorrhoidectomy (CH) and to define the role of SH in the treatment of hemorrhoids. DATA SOURCES: Published randomized controlled trials of CH vs SH with a minimum clinical follow-up of 12 months were searched and selected in the MEDLINE, EMBASE, and Cochrane Library databases using the keywords hemorrhoid, stapl, and anopexy, without language restrictions. STUDY SELECTION: Potentially relevant studies were identified by the title and the abstract, and full articles were obtained and assessed in detail. DATA EXTRACTION: Studies were scored according to the presence of 3 key methodologic features of randomization, blinding, and accountability of all patients, including withdrawals, and the scores ranged from 0 to 5. Studies that received a score from 3 to 5 were considered high-quality studies, whereas those with a score of 2 or less were considered of low quality. A specifically designed data form was used to collect all relevant data, including details of the experimental design, patient demographics, technical aspects, outcome measures, and complications. DATA SYNTHESIS: Fifteen articles met the inclusion criteria for a total of 1201 patients. Outcomes at a minimum of 1 year showed a significantly higher rate of prolapse recurrences in the SH group (14 studies, 1063 patients; odds ratio, 5.5; P < .001) and patients were more likely to undergo further treatment to correct recurrent prolapses compared with the CH group (10 studies, 824 patients; odds ratio, 1.9; P = .02). CONCLUSION: Stapled hemorrhoidopexy is a safe technique for the treatment of hemorrhoids but carries a significantly higher incidence of recurrences and additional operations compared with CH. It is the patient's choice whether to accept a higher recurrence rate to take advantage of the short-term benefits of SH. PMID- 19289668 TI - Total vs partial fundoplication in the treatment of gastroesophageal reflux disease: a meta-analysis. AB - OBJECTIVE: To perform a meta-analysis of randomized trials comparing partial fundoplication (PF) with total (Nissen) fundoplication (TF) for gastroesophageal reflux disease in terms of morbidity, efficacy, and long-term symptomatology. DATA SOURCES: A structured Medline search for published studies. STUDY SELECTION: The available literature from 1975 until June 2007 was searched using the Medical Subject Headings of the National Library of Medicine term fundoplication and the free-text terms fundoplication, surgery, and reflux. Data were analyzed using Review Manager software (Cochrane Collaboration, Oxford, England). DATA EXTRACTION: Eleven trials were identified comparing TF with PF in 991 patients. DATA SYNTHESIS: Total fundoplication resulted in a significantly higher incidence of postoperative dysphagia (odds ratio [OR], 1.82-3.93; P < .001), bloating (OR, 1.07-2.56; P = .02), and flatulence (OR, 1.66-3.96; P < .001). No significant differences were noted in the incidence of esophagitis (OR, 0.72-2.7; P = .33), heartburn (OR, 0.48-1.52; P = .58), or persisting acid reflux (OR, 0.77-1.79; P = .45). The reoperation rate was significantly higher after TF compared with PF (OR, 1.13-3.95; P = .02). No significant differences were present in the proportion of patients experiencing a good or excellent long-term outcome (OR, 0.54-1.38; P = .53) or in the proportion of patients with a Visick I or II score (OR, 0.62-1.59; P = .99). CONCLUSIONS: Partial fundoplication is a safe and effective alternative to TF, resulting in significantly fewer reoperations and a better functional outcome. The poor quality of the included trials warrants caution in the interpretation of the results of this meta-analysis. PMID- 19289669 TI - Asymptomatic isolated celiac artery dissection after a fall. AB - Isolated injury to mesenteric vessels in blunt trauma is uncommon. Most patients with these injuries present with abdominal pain, shock, or laboratory evidence of bowel and/or liver ischemia. We report herein the case of a man with asymptomatic isolated celiac artery dissection after blunt trauma suspected by screening abdominal computed tomography and confirmed by catheter-based angiography. The patient was treated with 3 months of oral anticoagulation alone. PMID- 19289670 TI - Image of the month. Adrenal myelolipoma. PMID- 19289671 TI - Image of the month. Gastrohepatic fistula following TACE for HCC. PMID- 19289672 TI - Image of the month. Left paraduodenal hernia. PMID- 19289673 TI - Total gastrectomy. PMID- 19289674 TI - Is laparoscopic colectomy a good operation for colon cancer? PMID- 19289675 TI - Voluntarism and the global unmet need for surgery. PMID- 19289676 TI - International surgical electives: intellectual capital, authorship, and capacity building. PMID- 19289677 TI - Long-term safety and efficacy of a new botulinum toxin type A in treating glabellar lines. AB - OBJECTIVE: To evaluate the long-term safety of repeated administrations of a new botulinum toxin type A (Reloxin; Medicis Pharmaceutical Corp, Scottsdale, Arizona) in the treatment of moderate to severe glabellar lines. METHODS: Open label assessment of 1200 patients receiving as many as 5 treatments of Reloxin over a 13-month period. The product was diluted in 2.5-mL sterile physiologic saline solution, 0.9%, without preservative to a concentration of 50 U of Reloxin per 0.25 mL of solution. Investigators injected 0.05 mL of the solution (10 U each) into each of 5 injection sites in the glabellar area on day 0 of each treatment cycle. There was a minimum 85-day gap between treatments. Postinjection clinical evaluation was performed on days 14 and 30 and monthly thereafter until retreatment, study completion, or early termination. The patients were telephoned on day 7 to check for adverse events (AEs) and concomitant medications, and patient diaries were used to document the onset of treatment effect. RESULTS: The majority (72%) of treatment-emergent AEs were considered unlikely or not related to study treatment. Probably or possibly related treatment-emergent AEs occurred in 36% of patients. The most frequently occurring related AEs were injection site disorders (18%), nervous system disorders (14% and 12% headache), and eye disorders (9%). Related AEs around the injection site or eyes were usually reported by day 7 and then resolved. Reported ptosis does not differentiate between brow ptosis and eyelid ptosis. A total of 45 patients had a total of 55 instances of ptosis across all cycles, with most episodes lasting less than 3 weeks. The rates of ptosis decreased during successive cycles from 2.4% in cycle 1 to 0.6% in cycle 5. The proportion of patients reporting an onset of response by day 7 ranged from 93% to 95%. By investigator assessment, the response rate (patients reporting none or mild glabellar line severity scale scores on day 30) ranged from 80% to 91% during cycles 1 to 5. CONCLUSIONS: There was no evidence of cumulative AEs or tachyphylaxis with multiple Reloxin treatments over a period of 13 months. The treatments were well tolerated. The rates of ptosis decreased over successive cycles, and the proportion of responders by day 7 ranged from 93% to 95%. PMID- 19289679 TI - Correction of the high female hairline. AB - OBJECTIVES: To review a technique and to make quantitative analyses of the senior author's 20-year experience with his preferred technique to correct the high female hairline. METHODS: A retrospective review of 29 female patients who underwent the hairline-lowering procedure performed by the same surgeon (S.S.K.). We analyzed preoperative and postoperative standardized photographs by taking measurements from the medial and lateral canthi to the anterior hairline. Facial height, from the menton to the hairline, was also measured. We calculated mean values and then used a 2-tailed, paired t test to evaluate for statistical significance. Patients also underwent evaluation for satisfaction, complications, and aesthetic result. We reevaluated the measurements from the profile view and compared them with the original data. RESULTS: The photographed midfrontal hairline position was vertically lowered on average 1.3 cm in patients who underwent a single-stage procedure (P < .001). In retrospect, the analysis was flawed compared with clinical experience. Therefore, the profile views were evaluated, and the correlating true curvilinear advancement was an average of 2.1 cm. Three complications occurred, including 1 major effluvium, 1 minor effluvium, and 1 scar that required revision. Patient satisfaction was extremely high. CONCLUSIONS: Advancement of the female hairline by incorporating an irregular trichophytic incision and a posterior scalp advancement flap is an effective and safe technique that has been used by the senior author for more than 2 decades. The average advancement was 2.1 cm in this study. The technique is immediately effective, well tolerated by patients, and associated with minimal complications. Although it is associated with a potentially visible incision, this technique can be used to make the scar virtually invisible. PMID- 19289680 TI - A contemporary assessment of facial aesthetic preferences. AB - OBJECTIVES: To compare and characterize the ideal nasolabial angle, nasal tip width, and location of the eyebrow apex for Asian and white women. METHODS: From approximately January 1, 2005, to June 30, 2005, we photographed the faces of 2 Asian women and 2 white women of differing ages. Each model's image was modified to create different eyebrow shapes, unique nasolabial angles, and varying nasal tip widths. We subsequently recruited, and obtained demographic information from, volunteers from the general public to rate the modified images based on their aesthetic preferences. RESULTS: We found that neither the ethnicity of the models nor the ethnicity of the volunteers who rated them played a significant role in determining the ideal eyebrow apex location, nasolabial angle, or nasal tip width. However, generally speaking, a more lateral brow apex is preferable in younger faces, whereas a more medial apex is favored in older ones. Other preferences include a moderate nasolabial angle and a narrow nasal tip. As a result of individual variability, it has been difficult to establish a method to calculate a nasolabial angle that adequately portrays the apparent rotation of the nose in most people. We found that the angle formed by the line from the anterior columella to the subnasale and the line exactly perpendicular to the Frankfurt horizontal plane provides the best estimate. CONCLUSIONS: When planning facial plastic surgery, the goals of the patient are of paramount importance. Although it is important to understand the ways in which people of different ethnicities and ages differ in their facial proportions as a group, facial harmony must be pursued on an individual basis. PMID- 19289681 TI - Pulsed-dye laser for treating ecchymoses after facial cosmetic procedures. AB - OBJECTIVE: To assess the safety and efficacy of a 595-nm pulsed-dye laser in the treatment of ecchymoses after facial cosmetic procedures. METHODS: Twenty consecutive patients with ecchymoses after facial cosmetic procedures underwent treatment with the pulsed-dye laser. A 10-mm spot size was used, with pulse duration of 6 milliseconds, fluence of 6 J/cm(2), and cryogen spray for 30 milliseconds with a 20-millisecond delay. The ecchymotic area was outlined; the lateral half was treated on postoperative day 5 or 6 and the medial half on postoperative day 7 to 10. Clinical photographs were obtained before and after each treatment. Three blinded independent observers evaluated the photographs and graded the ecchymoses on a scale of 0 to 3, with 3 indicating severe ecchymosis. RESULTS: The most common procedures associated with ecchymoses are cervicofacial rhytidectomy, facial lipocontouring, thread lift, and minimally invasive subperiosteal midface-lift. Pulsed-dye laser treatment resulted in a 63% mean improvement in ecchymosis scores within 48 to 72 hours. The only adverse effects were mild edema and discomfort. Maximal efficacy of the laser treatment was observed when it was performed between 5 and 10 days postoperatively. Patient satisfaction was universally high. CONCLUSIONS: Treatment with the pulsed-dye laser is safe and effective for expeditious resolution of postoperative ecchymoses after facial cosmetic procedures. It has the potential for wider application in treating postoperative ecchymoses on other areas of the body and after trauma. PMID- 19289682 TI - Polylactide-co-glycolide fiber-reinforced calcium phosphate bone cement. AB - OBJECTIVE: To compare the strength of polylactide-co-glycolide fiber-reinforced calcium phosphate bone cement (FRC) with nonreinforced calcium phosphate bone cement (NRC) subjected to simulated dural pulsations in defects larger than 25 cm(2). METHODS: Seven NRC and 7 FRC specimens were set in both medium (37.5 cm(2)) and large (50.0 cm(2)) model skull defects while subjected to simulated dural pulsations. Specimens were removed after 24 hours and analyzed using 3 point flexural testing. RESULTS: All 14 FRC specimens maintained structural integrity during extraction and testing. Only 2 of 7 (29%) medium specimens and 2 of 7 (29%) large NRC specimens survived setting. The mean (SD) energy to peak force (in newton millimeters [Nmm]) of the medium and large NRC specimens was 0.88 (0.83) and 3.00 (3.54) Nmm, respectively, compared with 28.97 (16.52) and 49.91 (38.10) Nmm for the medium and large FRC specimens. The material strength (in megapascals) of the medium and large NRC specimens was 0.17 (0.15) and 0.39 (0.33) MPa, respectively, compared with 3.73 (0.99) and 2.62 (1.34) MPa for the medium and large FRC specimens. The energy to peak force and material strength of the medium and large FRC specimens were significantly greater than for the corresponding NRC specimens; results were not statistically significant between medium and large FRC specimens. CONCLUSIONS: Fiber-reinforced calcium phosphate bone cement exhibits superior structural integrity and material strength than NRC when subjected to unshielded simulated dural pulsations. Further studies are needed to evaluate the biophysical parameters of FRC in vivo. PMID- 19289683 TI - Impact of vascular endothelial growth factor on skin graft survival in irradiated rats. AB - OBJECTIVE: To evaluate the effect of vascular endothelial growth factor (VEGF) on full-thickness skin graft (FTSG) survival on irradiated tissue as a model of wound healing in ischemic conditions. DESIGN: Twenty-four Sprague-Dawley rats underwent 30 Gy of irradiation to their left dorsum (10 Gy/d). After 4 weeks of recovery, 3-cm FTSGs were harvested from the healthy contralateral dorsum and placed onto irradiated recipient beds. Before grafting, recipient beds were delivered subfascial injections of either VEGF protein (5 microg) or physiologic saline. Graft failure (more than 10% necrosis) and graft microvascular density were compared between groups. RESULTS: Seven of the 11 FTSGs from saline-treated irradiated beds (64%) failed, whereas the failure rate for grafts treated with VEGF was 23% (3 of 13) (P = .048). Mean microvascular density was not different between groups. CONCLUSION: Exogenously administered VEGF may improve the outcome of FTSGs on irradiated tissue beds. PMID- 19289684 TI - Incidence of alarplasty in primary and revision rhinoplasty in a private practice setting. AB - OBJECTIVE: To evaluate the incidence of alarplasty in primary and revision rhinoplasty during a 3-year period. METHODS: We conducted a retrospective medical record review of patients undergoing primary and revision rhinoplasty between January 1, 2004, and December 31, 2006. Patients were divided into the following categories: primary or revision rhinoplasty, alarplasty, and ethnicity. The incidence of alarplasty was calculated and compared for primary vs revision rhinoplasty and for ethnicity. Statistically significant differences were confirmed using the chi(2) test. RESULTS: A total of 168 patients underwent rhinoplasty with minimum follow-up of 6 months; 116 patients had primary rhinoplasty and 52 revision rhinoplasty. All alarplasties were bilateral. Of 50 alarplasties performed, 35 were in the primary group and 15 in the revision group. The incidence of alarplasty was 30.2% in the primary group and 28.8% in the revision group. Nonwhite patients had a higher overall incidence of alarplasty (63.0%) than did white patients (25.4%). Each ethnic category identified had a higher incidence of alarplasty than the white group. CONCLUSIONS: Alarplasty is an important technique in rhinoplasty. The high incidence of alarplasty in revision rhinoplasty procedures suggests that alarplasty is underused by many surgeons currently performing rhinoplasty. One should consider alarplasty for all patients' noses, not just those of nonwhite ethnic rhinoplasty patients. PMID- 19289685 TI - Refining vertical lobule division in open septorhinoplasty. AB - OBJECTIVE: To review the indications, surgical techniques, and results of vertical lobule division (VLD) of the alar cartilages as they relate to the M Arch Model. DESIGN: Retrospective study of patients who underwent VLD of the lower lateral cartilages at a private facial plastic surgery practice in a major university teaching hospital. RESULTS: Vertical lobule division decreased projection in 34 of 41 patients, narrowed a wide or boxy tip in 25, corrected knuckling or bossae in 20, corrected tip asymmetry in 14, corrected a hanging columella in 14, increased rotation in 12, and decreased rotation in 6. No statistically significant correlation was noted between the location of VLD and the indication for which it was performed. One patient required revision surgery to increase rotation. CONCLUSIONS: Vertical lobule division remains a reliable and safe technique with predictable outcomes in tip repositioning. It allows for preservation of a strong tip complex, while adding versatility to tip refinement. PMID- 19289686 TI - Preserving structural integrity of the alar cartilage in aesthetic rhinoplasty using a cephalic turn-in flap. AB - Resection of the alar cartilage has long been a mainstay of aesthetic rhinoplasty. One drawback of this technique is the destabilization of the ala/lateral nasal wall complex. Herein we describe the cephalic turn-in flap, a technique for reinforcement of the alar cartilage after removal of its cephalic portion. PMID- 19289687 TI - A novel bioabsorbable device for facial suspension and rejuvenation. AB - To evaluate the safety and efficacy of a novel bioabsorbable suspension device made of a polymer of polylactic acid and polyglycolic acid (Endotine Ribbon), we performed a retrospective multi-institutional case study of 21 patients who underwent minimally invasive or open rhytidectomy with the use of the device in an ambulatory surgery center setting. Twelve patients had an excellent result, 7 a good result, and 2 a fair result. Early complications were corrected with technical modifications. Patient satisfaction was high. The Ribbon is a safe and effective adjunct for performing both minimally invasive and open rhytidectomy and cervical lifting. PMID- 19289688 TI - The lateral tarsal strip mini-tarsorrhaphy procedure. AB - The lateral canthus normally sits 1 to 2 mm higher than the medial canthus. With time, aging and gravity produce inferior displacement of the canthus. Numerous eyelid disorders can also result in lower eyelid or lateral canthal tendon laxity or malposition, requiring horizontal eyelid tightening or canthal repositioning. The lateral tarsal strip procedure has proven to be a useful technique in addressing these problems. Care must be taken when suspending the tarsal strip to the lateral orbital rim to preserve the almond shape of the lateral canthal angle. If mild to moderate upper eyelid laxity is present, suspension of the strip can result in upper eyelid overhang with lower eyelid and eyelash imbrication. We describe the lateral tarsal strip mini-tarsorrhaphy procedure that overcomes this problem. The technique provides excellent functional and aesthetic results and adds to the versatility of a time-tested procedure. PMID- 19289689 TI - Botulinum toxin-induced immobilization of lower facial wounds. PMID- 19289690 TI - Fatal septic thrombosis of the superior sagittal sinus after face-lift surgery caused by community-associated methicillin-resistant Staphylococcus aureus. PMID- 19289691 TI - High-volume calcium hydroxylapatite filler to the lower one-third of the face. PMID- 19289692 TI - Giovanni Boldini's Consuelo Vanderbilt (1876-1964), Duchess of Marlborough, and her son, Lord Ivor Spencer-Churchill (1898-1956). PMID- 19289693 TI - Neurodevelopmental outcome of appropriate and small for gestational age very low birth weight infants. AB - Prematurity and intrauterine growth restriction are associated with neurodevelopment delay. A prospective cohort study included all 96 very low birth weight infants discharged from our neonatal unit from November 2003 to June 2005. The authors compared 41 appropriate and 55 small for gestational age very low birth weight infants' neurodevelopment at 24 months corrected age. Bayley Scales were similar between both groups at 8, 12, 18, and 24 months corrected age even after adjustment for gestational age and improved from 8 to 24 months corrected age. Small for gestational age infants with and without severe intrauterine growth restriction had similar neurodevelopment outcome. Neurodevelopment delay is frequent in very low birth weight infants with no differences between appropriate and small for gestational age. The authors suggest that Bayley Scales evaluation should be performed in every very low birth weight infant at least at 24 months corrected age to detect those requiring therapeutic stimuli. PMID- 19289694 TI - Benign lesions accompanied by intractable epilepsy in children. AB - Epilepsy surgery has been proposed as a safe alternative treatment for intractable epilepsy in children, especially for patients with structural brain abnormalities. We studied 24 consecutive children who underwent surgery for intractable epilepsy. There were 12 males and 12 females. The mean age was 6.5 years. The seizures' duration ranged from 6 months to 2 years. The histopathological examination of the resected lesions revealed in 12 cases the presence of a ganglioglioma, in 7 cases dysembryoplastic neuroepithelial tumor, in 1 case a low grade glioma, in 2 cases cortical dysplasia, and in 2 cases cavernous malformations. In 18 cases, the lesions were located in the temporal lobe and in 6 cases the lesions were extratemporal. After a mean follow-up period of 4.4 years, 79% (19/24) of patients were seizure free. There were no permanent neurological deficits or deaths. Surgery for focal epilepsy in children is a safe procedure with favorable results. PMID- 19289695 TI - Temporary diazepam responsive apneic attacks and congenital myasthenic syndrome. AB - Congenital myasthenic syndromes are a genetically and phenotypically heterogeneous group of hereditary disorders affecting neuromuscular junction. Mutations in the gene encoding choline acetyltransferase cause presynaptic defects. The missense mutation I336T has been identified in Turkish population, and most of the cases carrying this mutation present with exercise-induced fatigability and ptosis. Although apneic attacks occur in these cases during febrile illness in childhood, the number of reported respiratory distress episodes during infancy is scarce. Another important feature of these cases is that response to esterase inhibitors is satisfactory. We present a case of congenital myasthenic syndrome with I336T choline acetyltransferase mutation who presented with numerous attacks of respiratory distress in the infancy period. Interestingly, the patient had myopathic findings on electromyography and diazepam decreased severity of apneic attacks. There was also no improvement with esterase inhibitors. PMID- 19289696 TI - Early signs of visual perception and evoked potentials in radiologically asymptomatic boys with X-linked adrenoleukodystrophy. AB - The aim was to identify the electrophysiological and psychological signs at a very early stage in asymptomatic boys with childhood cerebral X-linked adrenoleukodystrophy. Flash visual evoked potentials, pattern reversal, and visual event-related potentials were recorded in 6 radiologically asymptomatic boys with adrenoleukodystrophy and 22 control boys. The latency and amplitude of P100 of visual evoked potentials and P1 of event-related potentials were evaluated. Though all patients had normal intelligence quotient, performance intelligence quotient was significantly lower than verbal intelligence quotient in 2 patients. Both P100 and P1 amplitudes were significantly greater in adrenoleukodystrophy than in controls. The difference between performance intelligence quotient and verbal intelligence quotient exhibited significant correlation with P100 amplitude. Enlargement of visual evoked potentials might be a sign of cerebral involvement preceding the appearance of abnormalities on magnetic resonance imaging. Follow-up of asymptomatic boys with both electrophysiological and neuropsychological tests may serve as an aid for deciding the timing of therapeutic intervention. PMID- 19289697 TI - Clinical heterogeneity in ethylmalonic encephalopathy. AB - Ethylmalonic encephalopathy is a recently described inborn error of metabolism characterized clinically by developmental delay and regression, recurrent petechiae, orthostatic acrocyanosis, and chronic diarrhea. We describe monochorionic twins presenting with hypotonia in infancy and diagnosed with ethylmalonic encephalopathy on the basis of biochemical findings. They are compound heterozygote for missense mutations in ETHE1. Magnetic resonance imaging changes affecting the white matter, corpus callosum, and basal ganglia were seen in both patients. At 10 years of age, they have severe axial hypotonia but never displayed petechiae, orthostatic acrocyanosis, or chronic diarrhea. Their clinical courses differ markedly; one had an episode of coma when she was 3 years old and now has spastic quadraparesis and cannot speak. The other can freely use her upper extremities, her pyramidal syndrome being mostly limited to the lower extremities, and can speak 2 languages. These patients illustrate the clinical heterogeneity of ethylmalonic encephalopathy, even in monochorionic twins. PMID- 19289698 TI - Seizures in children following an apparent life-threatening event. AB - The characteristics of seizures and epilepsy in infants who have had an apparent life-threatening event have been poorly defined. Our objective was to characterize in depth the cohort of patients with apparent life-threatening events who developed seizures. We collected data from infants hospitalized for an apparent life-threatening event, and evaluated patients for subsequent seizures or chronic epilepsy. Of 471 patients with an apparent life-threatening event, 25 (5.3%) had seizures and 17 (3.6%) developed chronic epilepsy. There was no increased risk for febrile seizures. Abnormal brain magnetic resonance imaging results and developmental delay were only found in those patients who developed chronic epilepsy. Of those who developed chronic epilepsy, 47% were diagnosed with seizures within 1 week of their apparent life-threatening event. The discharge diagnosis at the time of the apparent life-threatening event was poorly predictive of those who developed seizures. In most cases the cause of chronic epilepsy was unknown, although cortical dysplasias made up a significant percentage (12%). PMID- 19289699 TI - Detection of bacteria in healthy middle ears during cochlear implantation. AB - OBJECTIVE: To assess whether free-living and/or biofilm bacteria are present in the putative sterile middle ear cavity before insertion of the electrode array during cochlear implantation. DESIGN: Prospective study. SETTING: Tertiary academic hospital. PATIENTS: The study included 45 healthy children (with or without a history of otitis media) undergoing cochlear implantation. INTERVENTIONS: Transmission electron microscopy or scanning electron microscopy was used to detect the presence of bacteria. MAIN OUTCOME MEASURE: Presence of both free-living bacteria and biofilm bacteria on the epithelial surface of biopsy specimens of middle ear mucosa. RESULTS: A majority of all mucosal specimens from clinically healthy tympanic cavities displayed inflammatory areas as well as dispersed, nonmatrix-enclosed bacteria. Also, rarely, fragments of biofilms were found. CONCLUSIONS: The presence of bacteria in the tympanic cavity, which is generally assumed to be sterile in healthy individuals, may provide an explanation for infectious complications after cochlear implantation. However, the possibility that the electrode array of a cochlear implant will actually become contaminated during insertion is unlikely because of the small amounts and dispersed presence of bacteria, which may account for the relatively low incidence of infectious complications after cochlear implantation. PMID- 19289700 TI - Auditory function after a prolonged course of ciprofloxacin-dexamethasone otic suspension in a murine model. AB - OBJECTIVE: To test for ototoxicity after prolonged ototopical fluoroquinolone use in the middle ear space using a murine model. DESIGN: Nonrandomized controlled trial. SUBJECTS: Twelve CBA/J mice. INTERVENTIONS: The mice received daily intratympanic injections of ciprofloxacin-dexamethasone otic suspension for 21 days. The contralateral ear received daily intratympanic injections of sterile isotonic sodium chloride solution (saline) as a control. MAIN OUTCOME MEASURES: Click-evoked auditory brainstem response (ABR) thresholds were obtained before injection and 10 days and 3 months after injection. RESULTS: Mean (SEM) preinjection ABR thresholds were 47.17 (2.74) dB peak equivalent sound pressure level (peSPL) in ciprofloxacin-dexamethasone-treated ears and 45.08 (1.56) dB peSPL in saline-treated ears (P = .38). Mean (SEM) postinjection ABR thresholds in ciprofloxacin-dexamethasone-treated ears were 44.25 (1.25) dB peSPL after 10 days and 43.00 (1.51) dB peSPL after 3 months. Mean (SEM) postinjection ABR thresholds in saline-treated ears were 48.00 (1.51) dB peSPL after 10 days and 45.92 (1.79) dB peSPL after 3 months. There were no significant differences in ABR thresholds for ciprofloxacin-dexamethasone-treated ears (P = .29, P = .10) or saline-treated ears (P = .07, P = .59). CONCLUSION: Ciprofloxacin-dexamethasone suspension did not cause either immediate or delayed ototoxicity after 21 days of intratympanic administration in the CBA/J mouse model. PMID- 19289701 TI - Polymerase chain reaction for pathogen identification in persistent pediatric cervical lymphadenitis. AB - OBJECTIVE: To study routine culture-negative persistent cervical lymphadenitis in children treated surgically during a 10-year period (December 26, 1997, to October 1, 2007) at a single institution. DESIGN: Retrospective case series. SETTING: Tertiary university-based pediatric referral center. PATIENTS: Patients 18 years or younger with cervical lymphadenitis managed surgically (incision and drainage, curettage, and/or excisional lymphadenectomy) and medically (antibiotic therapy), culture-negative after 48 hours, and subsequently diagnosed using the polymerase chain reaction, extended culture incubation, and/or histopathologic evaluation. MAIN OUTCOME MEASURES: Number of surgical interventions, causative organisms, histopathologic features, and resolution of lymphadenitis. RESULTS: Ninety surgical procedures were performed in 60 patients. The cure rate was 23% (approximately 14 patients) with incision and drainage, 58% (approximately 35 patients) with curettage, and 95% (57 patients) with excisional lymphadenectomy. Nontuberculous mycobacteria were the most prevalent causative organisms, followed by Bartonella and Legionella organisms. Four of 6 patients with Bartonella infection had a history of cat exposure, and 4 of 6 patients with Legionella infection had a history of hot tub exposure. CONCLUSIONS: Excisional lymphadenectomy is the preferred treatment of mycobacterial persistent cervical lymphadenitis in children. Sufficient data are lacking for similar recommendations in patients with disease caused by Bartonella organisms, whereas for neck disease caused by Legionella organisms, excisional lymphadenectomy may be superior to incision and drainage. The polymerase chain reaction is useful for pathogen identification in pediatric cervical lymphadenitis, although it is less sensitive in identification of mycobacteria. To our knowledge, our study is the first to report multiple cases of legionellosis in otherwise healthy children. Legionella seems to be a previously unrecognized but relatively common pathogen in culture-negative persistent cervical lymphadenitis in children. PMID- 19289702 TI - High-resolution ultrasound in the evaluation of pediatric recurrent respiratory papillomatosis. AB - OBJECTIVES: To characterize the ultrasonographic appearance of laryngeal papillomatosis and to compare ultrasound with direct laryngoscopy and bronchoscopy, the criterion standard, for airway evaluation. DESIGN: Prospective, nonrandomized analysis of preoperative and postoperative airway ultrasound images. SETTING: Tertiary, university-based medical center. PARTICIPANTS: Eight patients (4 females and 4 males) with recurrent respiratory papillomatosis, with a mean age of 10.25 years and a mean of 14 surgical papilloma resections (range, 3-35). INTERVENTION: The patients underwent planned papilloma resections with ultrasound evaluation before formal endoscopic resection. Preresection ultrasound images of respiratory papillomas were evaluated. MAIN OUTCOME MEASURES: The ultrasonographic appearance of respiratory papillomas and pediatric airway anatomy. RESULTS: Respiratory papillomas appeared as discrete, hyperechoic lesions on the relatively hypoechoic background of the true vocal folds. CONCLUSIONS: Recurrent respiratory papillomas have a characteristic ultrasonographic appearance that seems to correlate with endoscopic findings. It seems that this modality holds promise for identifying pedunculated papillomas. Although direct laryngoscopy and bronchoscopy are the criterion standard, airway ultrasound may have a role in the early diagnosis of, surveillance of, and operative planning for recurrent respiratory papillomatosis. To our knowledge, this is the first study to describe the ultrasonographic appearance of papillomas and the first comparison of ultrasonographic and endoscopic airway images. This modality merits further study, and further investigation is ongoing. PMID- 19289703 TI - Downregulation of TESTIN and its association with cancer history and a tendency toward poor survival in head and neck squamous cell carcinoma. AB - OBJECTIVE: To examine the role of TESTIN as a candidate tumor suppressor gene in head and neck carcinogenesis. DESIGN: Mutation and messenger RNA (mRNA) expression analyses. SETTING: Academic research. PATIENTS: Paired normal and tumor samples were obtained from 38 patients with primary head and neck squamous cell carcinoma. MAIN OUTCOME MEASURES: Analysis and comparison of TESTIN gene mRNA expression and its relationship to clinicopathologic variables. RESULTS: Mutation analysis showed a nucleotide and amino acid change in 6 of the 38 tumor samples (16.0%). Semiquantitative mRNA expression analysis of TESTIN revealed a decreased expression in approximately 50% of the tumors compared with their matched normal controls. Interestingly, comparison of clinicopathologic variables to mRNA expression status of TESTIN revealed a significant difference in terms of cancer history (P = .03). Moreover, a higher smoking ratio and a family cancer history were also associated with downregulation of TESTIN, although the difference was not statistically significant (P = .43 and P = .16, respectively). Kaplan-Meier survival analysis demonstrated a worse survival rate among the patients with low TESTIN expression compared with the patients with normal-high TESTIN expression. CONCLUSIONS: Our findings suggest that inactivation of TESTIN is involved in head and neck carcinogenesis through its downregulation. Further studies in various human cancer tissues using a large sample size and in vitro functional studies as well as clinical comparison research studies would give us a better evaluation of TESTIN's role and its possible future application in molecular diagnosis and treatment of different cancer types, including head and neck squamous cell carcinoma. PMID- 19289704 TI - 7beta-hydroxycholesterol induces apoptosis and regulates cyclooxygenase 2 in head and neck squamous cell carcinoma. AB - OBJECTIVE: To determine whether treatment with 7beta-hydroxycholesterol (7beta HC) would trigger cell death in head and neck squamous cell carcinoma (HNSCC) cell lines in a dose-dependent fashion. DESIGN: In vitro study. SUBJECTS: The study included HNSCC cell lines SCC9, SCC25, CAL27, and FaDu. INTERVENTION: We treated HNSCC cell lines with increasing doses of 7beta-HC. Proliferation assays were performed to assess cell viability after treatment. Western blots were carried out to evaluate cyclooxygenase (COX)-1 and -2 expression levels. RESULTS: Using proliferation assays and immunocytochemical analysis, we detected significant growth inhibition via apoptosis in 4 different HNSCC cell lines after treatment with 7beta-HC (P < .001). The 50% inhibitory concentration levels were between 13.19 and 20.79 micromol/L after 72 hours. Western analysis indicated that COX-2, but not COX-1, levels were suppressed after treatment. CONCLUSIONS: Treatment with 7beta-HC resulted in suppression of HNSCC growth in vitro. Our data warrant further investigations for the potential use of 7beta-HC as a cytotoxic agent in head and neck cancer. PMID- 19289705 TI - Technical considerations in patients requiring a second microvascular free flap in the head and neck. AB - OBJECTIVE: To identify the difficulties associated with performing a second free flap reconstruction in the head and neck as well as the techniques used to achieve successful reconstruction. DESIGN: A retrospective review of a prospectively maintained patient database. PATIENTS: The study population comprised 33 patients who required a second free flap in the head and neck. MAIN OUTCOME MEASURES: Several variables were analyzed in this cohort. Patient-related factors included the following: the availability of recipient vessels, the need to access the contralateral side of the neck, history of neck surgery, history of radiation therapy, and type of free flap used. Complications associated with the reconstruction were also reported. RESULTS: In this cohort of 33 patients, 34 free flap reconstructions were performed. All were successful, and there were no flap-related complications. Owing to the paucity of good recipient vessels in many cases, the contralateral side of the neck was commonly used, but no patients required vein interposition grafts. CONCLUSIONS: Second free flap reconstructions in the head and neck can be safely accomplished. We have found that the availability of recipient vessels is the most important consideration in these patients. The dissection of the contralateral side of the neck is often required, but the use of flaps with long pedicles obviates the need for vein interposition grafts. Compared with the success rate in this study, the relevant literature suggests that flap survival rates are lower when interposition grafts are used. Modifications in flap-harvesting techniques and inset geometry can also be used to facilitate insets in complicated surgical fields that have been reoperated on. PMID- 19289706 TI - High-speed laryngeal imaging compared with videostroboscopy in healthy subjects. AB - OBJECTIVE: To describe normal vocal fold vibratory characteristics recorded with high-speed digital imaging (HSV) of the larynx. DESIGN: Prospective study of healthy subjects who volunteered to undergo laryngeal HSV and videostroboscopy. Image analysis was randomly assigned to 3 blinded raters. SETTING: Community based clinic with a specialty in laryngology. PARTICIPANTS: Fifty healthy subjects aged 21 to 65 years who were nonsmokers and who had no voice problems, laryngopharyngeal reflux, or reactive airway disease. MAIN OUTCOME MEASURES: The following characteristics of vibration were described: glottal configuration, phase closure, vibratory symmetry, mucosal wave propagation, amplitude of vibration, and periodicity of vibration. Interrater and intrarater reliabilities were calculated for both imaging modalities. RESULTS: The range of findings for each measure is described. The comparison of videostroboscopy ratings with ratings from HSV studies did not reveal any significant difference between the 2 modalities for any of the measures except for the assessment of periodicity. Aperiodic vibratory characteristics were noted on 30% of the videostroboscopy studies (n = 15) and in only 4% of the HSV studies (n = 2) (P <. 001). Although interrater and intrarater agreement were considered to be generally acceptable, a significant rater effect was identified. CONCLUSION: This preliminary study describes a range of normal values for vocal fold vibratory characteristics as recorded with laryngeal HSV, providing a basis for comparison of studies in patients with voice problems. PMID- 19289707 TI - Cyclooxygenase 2 promoter-based replication-selective adenoviral vector for hypopharyngeal cancer. AB - OBJECTIVE: To explore the potential clinical application of the oncolytic activity of cyclooxygenase 2 (COX-2) promoter-based, conditional, replication selective adenovirus vector for hypopharyngeal squamous cell carcinoma. DESIGN: In vivo study and retrospective study. SETTING: Kobe University Hospital, Kobe, Japan. SUBJECTS: Expression of COX-2 in hypopharyngeal cancers treated at Kobe University Hospital was immunohistochemically investigated. In addition, nude mice bearing human hypopharyngeal cancer cells (H891) were used to analyze oncolytic activity of a conditional replication-selective adenovirus vector in which the expression of E1a, required for viral replication, is controlled by the COX-2 promoter Ad-COX2-E1a. RESULTS: In vivo assays showed significant growth suppression in the murine hypopharyngeal model. Cyclooxygenase 2 expression was observed in 75.3% of hypopharyngeal cancers, especially in differentiated tumor cells (P = .001; r = 0.433). CONCLUSION: In this study, we demonstrated the potential of oncolytic therapy using the COX-2-promoter based, conditional, replication-selective adenovirus for COX-2-expressing hypopharyngeal squamous cell carcinomas. PMID- 19289708 TI - Repeated botulinum toxin type A injections to treat patients with Frey syndrome. AB - OBJECTIVE: To determine the effectiveness of repeated intracutaneous injections of botulinum toxin A to treat Frey syndrome. PATIENTS: Between January 6, 1999, and July 1, 2005, 22 patients with Frey syndrome (12 men and 10 women) received repeated intracutaneous injections of botulinum toxin type A. MAIN OUTCOME MEASURES: Before each treatment, the time since the previous treatment; the size of the affected area, as determined by a starch-iodine test; and subjective quantification of symptoms, as assessed by the previously used Frey Questionnaire Card, were recorded. RESULTS: All patients underwent at least 3 treatments. Univariate analysis of variance showed a significant difference (P <.001) in the between-treatment interval. Univariate analysis of variance showed a significant difference in the size of the affected area in accordance with the number of treatments received (P <.001). The mean Frey Questionnaire Card score also decreased with repeated treatments. CONCLUSIONS: In patients with Frey syndrome, repeated treatment with intracutaneous injection of botulinum toxin type A lowered subjective symptom scores, decreased the size of the affected area, and increased the duration of the effect. PMID- 19289709 TI - Effects of pentoxifylline on olfactory sensitivity: a postmarketing surveillance study. AB - OBJECTIVE: To investigate the effect of pentoxifylline, an unspecific phosphodiesterase inhibitor, on olfactory function. DESIGN: Longitudinal study. PATIENTS: Nineteen patients who received pentoxifylline to treat inner-ear conditions. MAIN OUTCOME MEASURES: Evaluation of olfactory function (ie, odor threshold, odor discrimination, and odor identification) before and after administration of pentoxifylline and assessment of nasal airflow. RESULTS: Administration of pentoxifylline had no significant effect on nasal airflow (P = .84). After administration of pentoxifylline, patients demonstrated a decrease in odor threshold toward lower odor concentrations (P = .01). The odor threshold shift after administration of pentoxifylline was more pronounced in younger patients than in older patients (correlation between age and change in odor threshold: r = -.72; P = .001). No such changes were observed for suprathreshold olfactory tasks (odor discrimination and odor identification). Six of the 19 patients were found to have hyposmia. Two patients demonstrated a clinically significant decrease in odor threshold after drug administration. CONCLUSIONS: The present findings may indicate a role for pentoxifylline in the treatment of olfactory loss. Double-blind, placebo-controlled studies are needed to verify whether pentoxifylline can improve olfactory sensitivity in patients with olfactory disorders. PMID- 19289710 TI - Use of mometasone furoate to prevent polyp relapse after endoscopic sinus surgery. AB - OBJECTIVE: To evaluate the effect of mometasone furoate on prevention or reduction of nasal polyp relapse and worsening of symptoms after functional endoscopic sinus surgery (FESS). DESIGN: Randomized, double-blind, placebo controlled, multicenter study. SETTING: Ten ear, nose, and throat clinics in Sweden. PATIENTS: Adult subjects with bilateral nasal polyps fulfilling the criteria for surgery who underwent FESS. INTERVENTIONS: Two weeks after FESS, subjects were randomized to receive mometasone furoate nasal spray, 200 microg once daily, or placebo. MAIN OUTCOME MEASURES: Time to relapse, defined as an increase of 1 point or more on a 0- to 6-point endoscopic polyp scale. RESULTS: In the per-protocol population (n = 104), median time to relapse was 173 and 61 days for the mometasone and placebo groups, respectively (P = .007; hazard ratio [95% confidence interval], 0.72 [0.55-0.93]). In the intent-to-treat population (n = 159), median time to relapse was greater than 175 days in the mometasone group and 125 days in the placebo group (P = .049; hazard ratio, 0.79 [0.62 0.99]). The most common adverse event was epistaxis, with 6 cases reported in the mometasone group and 3 in the placebo group. CONCLUSIONS: Postoperative use of mometasone furoate, 200 microg once daily, provided a statistically significant longer time to relapse of nasal polyps than did placebo in subjects with bilateral nasal polyposis who had undergone FESS. The ability of mometasone to prevent or prolong the time to relapse among subjects undergoing FESS is important because this may prolong the time to subsequent surgery. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00731185. PMID- 19289711 TI - The effect of nasally administered budesonide respules on adrenal cortex function in patients with chronic rhinosinusitis. AB - OBJECTIVES: To evaluate whether nasal administration of budesonide in adults with chronic rhinosinusitis for 30 days suppresses adrenal function and to assess its clinical efficacy. DESIGN: An open-label prospective study. SETTING: Academic medical center. PATIENTS: We assessed adrenal function in 9 patients using the cosyntropin test before and after budesonide therapy. INTERVENTION: Budesonide respule therapy. MAIN OUTCOME MEASURE: Scores from the Sino-Nasal Outcome Test-20 (SNOT-20), a tool for assessing rhinosinusitis health and quality of life, were used to assess efficacy of budesonide treatment. RESULTS: All of our patients showed adequate adrenal response to cosyntropin stimulation before and after the budesonide trial. The mean difference in SNOT-20 scores was -1 (95% confidence interval, -1.77 to -0.23; P = .02), indicating clinically significant improvement after therapy. CONCLUSION: Our findings suggest that using budesonide nasal wash may be clinically effective in decreasing the symptoms of chronic rhinosinusitis and does so without suppression of the hypothalamic-pituitary-adrenal axis in patients with chronic rhinosinusitis. PMID- 19289712 TI - Lingual osteoma: case report and literature review. PMID- 19289713 TI - Endoscopic management of idiopathic spontaneous skull base fistula through the clivus. PMID- 19289714 TI - Radiology quiz case 1. Massive cervicothoracic hematoma due to extracapsular rupture of a parathyroid adenoma. PMID- 19289715 TI - Radiology quiz case 2. Extranodal Rosai-Dorfman disease involving the right orbit and nasal cavity. PMID- 19289716 TI - Pathology quiz case 1. Salivary gland anlage tumor (SGAT). PMID- 19289717 TI - Pathology quiz case 2. Chronic sclerosing sialadenitis (CSS). PMID- 19289718 TI - Issues in the interpretation and reporting of surveys in dental education. AB - Surveys are the most common form of data-based article published in the Journal of Dental Education. The apparent ease with which they are conducted and the seeming simplicity of reporting results mask significant issues in sample design and performing maximally useful analyses. Four concerns are discussed here. First, it is demonstrated that results are a function of who, when, and where responses are sampled, each source making independent contributions. Second, absolute sample size is shown to be the most significant factor affecting precision in surveys, and the numbers of schools, respondents, and other sources of variance can be chosen to minimize survey imprecision. Third, response rate typically has negligible effect on precision and an uncertain effect on accuracy (freedom from bias). A technique, sample saturation, is explained that can be used to protect, to some degree, surveys from the effects of bias. Finally, suggestions are offered for reporting survey results in a visually meaningful fashion, and an appeal is made that recommendations associated with surveys not be published unless they are grounded in both data and well-developed theory. This analysis references a previously published survey on competency-based dental education to illustrate methodological points in concrete terms. PMID- 19289719 TI - Impact of community-based oral health experiences on dental students' attitudes towards caring for the underserved. AB - Dental students' attitudes towards access to dental care for the underserved may be impacted by participation in community oral health promotion programs that target individuals in underserved communities. At the University of Southern California School of Dentistry, freshman dental students provide classroom oral health promotion and preventive dental care programs to underserved elementary school children. One hundred forty-four freshman dental students were surveyed three times during their freshman year-before, during, and after participation in these programs. The students' attitudes about societal expectations, health professionals' responsibility, access to care, and students' personal efficacy to positively impact the need for expanded oral health care services for segments of the population were measured. Students reported positive attitudes in all categories throughout the study period. The students' attitudes about societal expectations to care for the oral health of the underserved remained stable over the study period, but they became more uncertain of who should be responsible for fulfilling that obligation, who should receive that care, and their capability to provide this care while in dental school. These changes in attitude may reflect the students' greater understanding of the complexity of the determinants of oral health as a consequence of their community education experiences. PMID- 19289720 TI - Tomorrow's leaders, starting today: a pilot leadership development program for dental students. AB - Effective leadership is vitally important as the dental profession strives to meet current and future challenges. Leadership development programs have been created for mid-career dental professionals, but the relative lack of such programs for dental students may represent a missed opportunity to cultivate the dental leaders of tomorrow. A pilot leadership development program for dental students is described in this article. A voluntary leadership development program for dental students was offered in 2008 at the Case School of Dental Medicine with support from the Ohio Dental Association Foundation. The program aimed to increase students' leadership knowledge, improve their leadership skills, and provide inspiration through exposure to leaders who could serve as role models. At the conclusion of the program, students attended the Ohio Dental Association's Leadership Institute event. Forty-six students attended at least one program session. Thirty students attended all or all but one of the on-site sessions. Thirty-three participants responded to a post-program anonymous online survey. The majority of participants (81 percent) rated the program as very useful or useful and said they would participate in the program again (85 percent). Student attendance at the state dental association's leadership event increased appreciably from previous years. Student participation in the pilot program exceeded expectations. Leadership development programs for dental students are feasible and can benefit students and the dental community. PMID- 19289721 TI - Learning styles of orthodontic residents. AB - Significant challenges face many orthodontic residency programs, particularly a shortage of full-time experienced faculty members. Due to this shortage, it is critical that program directors design comprehensive curricula that incorporate the most effective and efficient teaching methods. It is theorized that teaching effectiveness and efficiency are optimized when the course design and content closely match students' learning preferences. This survey study was designed to distinguish the learning preferences of orthodontic residents utilizing Felder and Soloman's Index of Learning Styles, which assesses student learning preferences in four dimensions using dichotomous scales, thereby providing insight into how teaching strategies can best be structured. As a secondary focus, additional questions on the survey were asked to gain information about residents' access to the Internet and comfort level with online learning so as to address acceptance of web-based courses in response to the shortage of full-time faculty members. Orthodontic residents, contacted via email, were requested to complete an online survey; 261 responses were collected. The results indicate that orthodontic residents are highly visual learners and show a preference for sensing and sequential learning strategies. In terms of information technology, the residents are comfortable with and have adequate access to current technological assets; therefore, they may be well suited for inclusion of computer-based teaching modules and other multimedia devices in their residency curriculum. PMID- 19289722 TI - A comparative study of professional student stress. AB - A study was conducted involving a group of 290 medical and dental students to directly compare perceived stress levels encountered during their education. A modified questionnaire based on Garbee et al.'s Dental Environmental Stress survey was provided to the students by either email or paper. The purpose of the investigation was to determine if the sources of stress reported by medical and dental students, both male and female, were due to common factors. A multivariate statistical analysis was also conducted to measure stress differences by year in school. Through factor analysis, the survey question responses were grouped into five causal categories: academic performance, faculty relations, patient and clinic responsibilities, personal life issues, and professional identity. The overall findings show that dental students had greater levels of stress than medical students in three of the five categories. The only category in which medical students demonstrated greater stress levels than dental students was in professional identity. Measures of comparative levels of stress between male and female students for either profession did not demonstrate any significant differences. Stress levels related to clinical work varied significantly between the type of professional student and his or her year in school. PMID- 19289723 TI - Factors influencing pediatric dental program directors' selection of residents and demographics of current directors. AB - The primary goal of this study was to examine the criteria that influence rankings of candidates by advanced education program directors in pediatric dentistry. Secondary objectives were to obtain information on the resident selection process and to explore demographics of current program directors. A survey was sent in 2005 to all sixty-three program directors of pediatric dentistry residency programs accredited within the United States for the graduating class of 2007. The survey had a response rate of almost 78 percent (49/63). Respondents were requested to rank the importance of eleven factors that are typically included in the selection criteria for pediatric dentistry residents. Factors were rated on a scale of critical, very important, fairly important, somewhat important, and not important. The four highest ranked criteria by program directors were the following, in order: National Board scores, dental school clinical grades, class rank, and grade point average (GPA). Other factors ranked in descending order of perceived importance were the following: dental school basic science grades, experience in pediatric dentistry, extracurricular activities, completion of a general practice residency or advanced education in general dentistry program, the application essay, a publication or professional presentation, and private practice experience. All directors ranked personal interviews as very important to critical. Letters of recommendation from a pediatric dentistry department chairperson or faculty member were viewed more favorably than letters from dental school deans and non pediatric dentistry faculty. Fifty-seven percent of the directors responding (28/49) were male, and 81 percent (40/49) were white, non-Hispanic. Fifty-nine percent of the directors (29/49) graduated from a residency program over twenty years ago, with 39 percent (19/49) having been a director for less than five years. PMID- 19289724 TI - Dental hygiene students' attitudes toward ethical dilemmas in practice. AB - This article reports the findings of a survey-based study conducted in 2006 to determine graduating dental hygiene students' attitudes toward ethical dilemmas in eight areas of practice: substandard care, overtreatment of patients, scope of practice, fraud, confidentiality, impaired professionals, sexual harassment, abuse, and health status. The findings, based on responses from 1,165 students at 141 U.S. dental hygiene programs, indicate that many dental hygiene students do not understand what behaviors in the patient care environment are consistent with ethical practice and which are not. Responding students believed that hygienists have a strong duty to report, intercede, or educate in areas of abuse, sexual harassment, detection of cancer, and smoking cessation. However, they were less likely to report concerns about ethical transgressions such as fraud, inadequate infection control, exceeding practice scope, and failure to diagnose disease when such disclosures could potentially threaten their employment status. Based on the results, we recommend that dental hygiene programs explore curriculum enhancements to improve students' comprehension of what constitutes fraud and other ethical transgressions and the proper reporting mechanisms. PMID- 19289725 TI - Utilizing public health clinics for service-learning rotations in dental hygiene: a four-year retrospective study. AB - National reports outlining disparities in oral health care in the United States have focused attention on ways to encourage health care providers to become more involved in the public health arena. Utilization of service-learning in professional health education programs is one method being explored. The purpose of this study was to conduct a retrospective review of a service-learning rotation within a dental hygiene public health course. The study utilized data sources generated by students as part of a course evaluation. These sources included student journals (qualitative/quantitative) and Likert-scaled (quantitative) and open-ended (qualitative) student satisfaction survey items. Mixed methodology data analysis techniques were used to analyze and triangulate data in order to form conclusions related to the effectiveness of service learning as a teaching strategy in dental hygiene. This investigation suggests that service-learning is an effective learning strategy for increasing student awareness of underserved populations, cultural diversity, and ethical patient care. The study also suggests that service-learning helped students to determine their level of interest in public health as a career choice by giving them a real world experience in public health patient care. PMID- 19289726 TI - An assessment of promotion and tenure requirements at dental schools. AB - Tenure and promotion are integral parts of the academic environment; however, in this era of growing shortages of faculty, as the baby boomers retire and the majority of dentists choose private practice, recruitment and retention may be affected by the way institutions handle promotion and tenure. A national survey of U.S. dental schools conducted in 2007 assessed the existence of multiple employment tracks for faculty and examined the requirements for promotion and tenure. All responding schools reported the existence of multiple tracks for academic faculty appointments. Many dental schools reported that they provided opportunity for faculty members to switch from one track to another, thus circumventing the traditional "up-or-out" policy. The number of schools offering non-tenure appointment tracks has also increased. This finding indicates an apparent increase over time in flexibility regarding tracks and tenure. The majority of schools did not report requirements for the number of publications necessary for promotion. Those that did report requirements showed an increase in expected scholarly activity relative to past studies, indicating that it may be more difficult than ever to achieve promotion. PMID- 19289727 TI - Dental hygiene faculty calibration in the evaluation of calculus detection. AB - The purpose of this pilot study was to explore the impact of faculty calibration training on intra- and interrater reliability regarding calculus detection. After IRB approval, twelve dental hygiene faculty members were recruited from a pool of twenty-two for voluntary participation and randomized into two groups. All subjects provided two pre- and two posttest scorings of calculus deposits on each of three typodonts by recording yes or no indicating if they detected calculus. Accuracy and consistency of calculus detection were evaluated using an answer key. The experimental group received three two-hour training sessions to practice a prescribed exploring sequence and technique for calculus detection. Participants immediately corrected their answers, received feedback from the trainer, and reconciled missed areas. Intra- and interrater reliability (pre- and posttest) was determined using Cohen's Kappa and compared between groups using repeated measures (split-plot) ANOVA. The groups did not differ from pre- to posttraining (intrarater reliability p=0.64; interrater reliability p=0.20). Training had no effect on reliability levels for simulated calculus detection in this study. Recommendations for future studies of faculty calibration when evaluating students include using patients for assessing rater reliability, employing larger samples at multiple sites, and assessing the impact on students' attitudes and learning outcomes. PMID- 19289728 TI - Primer on preclinical instruction and evaluation. AB - This review summarizes the available literature for both instruction and evaluation of the novice dental and dental hygiene preclinical student. Effective instruction for dental and dental hygiene instrumentation requires knowledge of motor skills theory and mechanisms of fine motor skills attainment. The novice learner requires small, explicit steps that clearly define production. Prior to any performance, the skill to be performed should be envisioned accurately by the learner. Timely, precise feedback from the instructor to the learner contributes to learning. Novices are unable to judge their performance accurately, so self assessment skills must be taught. Repetition enhances motor performance. Instruction is supported through well-designed evaluation instruments containing explicit criteria arranged in the correct order of production. Assessment tools should be designed to aid in providing specific, immediate feedback. Well designed assessments may also aid in calibration of instructors. Examples of evaluation instruments are found in the literature, and several are reviewed in this article. For those responsible for preclinical performance assessment, application of current motor skills theory and development of appropriate instruction and evaluation instruments may result in improved student performance. Studies also indicate the instructional environment in the dental clinical setting may be less stressful if evaluation is based on achievement of target levels rather than multiple daily grades. PMID- 19289729 TI - Advanced placement, qualifying, and degree completion programs for internationally trained dentists in Canada and the United States: an overview. AB - Canadian and U.S. universities are faced with the challenge that they are not graduating enough dentists to meet the future needs of the Canadian and U.S. populations. Foreign-trained dentists represent a valuable resource to society and the economy. Dental programs have been established to train foreign-trained dentists for some or all of the following reasons: public need for health care services, income generation for universities, and demand by foreign-trained dentists who desire to practice dentistry in Canada or the United States rather than in their own nation. Changes implemented by the National Dental Examining Board (NDEB) of Canada in 2000 and by the Commission on Dental Accreditation (CODA) in the United States in 1986 have resulted in foreign-trained dentists no longer being able to gain dental licensure in these countries strictly through a certification examination. Foreign-trained dentists are now required to apply for and complete a two- to three-year advanced placement, qualifying, or degree program at a Canadian or U.S. dental school prior to receiving licensure to practice. The study reported in this article investigated the various types of advanced placement, qualifying, or degree programs available to foreign-trained dentists wishing to practice in either Canada or the United States and the differences among these programs. This research provides a better understanding of the various commonalities and differences among Canadian and U.S. programs for internationally trained dentists. PMID- 19289730 TI - Patterns of dental therapists' scope of practice and employment in Victoria, Australia. AB - In Australia, dental therapists have practiced only within the state-operated School Dental Services (SDS) for around forty years providing preventive, diagnostic, restorative, and health promotion services to children and adolescents in a collaborative and referral relationship with dentists. Changes to legislation in 2000 have seen limits to dental therapists' employment removed, allowing private sector employment. This study examines the changes to dental therapists' employment since 2000 using a self-completed questionnaire with a response rate of 82 percent. Approximately one-third of responding dental therapists reported that they spent some time employed outside the SDS in community health services and private orthodontic and general practices, which indicates an acceptance of this type of dental care provider in these areas. The clinical services that dental therapists are currently providing are a complex mix with significant variations according to type and geographical location of practice, but include high levels of patient assessment, diagnosis, treatment planning, and the restoration of teeth. The findings from this study indicate that when legislative restrictions on employment settings are removed, there is a demand and demonstrable role for dental therapist-delivered services in nongovernment dental practices. PMID- 19289731 TI - Teaching NeuroImages: face of the giant panda and her cub: MRI correlates of Wilson disease. PMID- 19289732 TI - Emerging subspecialties in neurology: neuro-oncology: a developing subspecialty with many opportunities. PMID- 19289734 TI - The numb and the restless: peripheral neuropathy and RLS. PMID- 19289735 TI - Sensitivity vs specificity: progress and pitfalls in defining MRI criteria for pediatric MS. PMID- 19289736 TI - A splice site variant in the sodium channel gene SCN1A confers risk of febrile seizures. AB - OBJECTIVE: Our aim was to investigate whether the risk of febrile seizures is influenced by a common functional polymorphism in the sodium channel gene SCN1A. This single nucleotide polymorphism (IVS5N+5 G>A, rs3812718) was shown to modify the proportion of two alternative transcripts of the channel. METHODS: We performed an exploratory case-control association analysis in 90 adult epilepsy patients with childhood febrile seizures vs 486 epilepsy patients without a history of febrile seizures and also vs 701 population controls. In the replication step, we investigated children with febrile seizures without concomitant epilepsy at the time of their inclusion. We compared the genotypes of 55 of those children against population controls and performed a within-family association analysis in an additional 88 child-parent trios with febrile seizures. RESULTS: We observed a significant association of the splice-site interrupting A-allele with febrile seizures (p value in the exploratory step: 0.000017; joint p value of the replication: 0.00069). Our data suggest that the A allele of this variant confers a threefold genotype relative risk in homozygotes and accounts for a population attributable fraction of up to 50% for the etiology of febrile seizures. CONCLUSIONS: The A-allele of the SCN1A single nucleotide polymorphism IVS5N+5 G>A (rs3812718) represents a common and relevant risk factor for febrile seizures. A limitation of the present study is that patients of the exploratory and replication steps differed in aspects of their phenotype (febrile seizures with and without additional epilepsy). PMID- 19289737 TI - High-frequency oscillations mirror disease activity in patients with epilepsy. AB - OBJECTIVE: High-frequency oscillations (HFOs) can be recorded in epileptic patients with clinical intracranial EEG. HFOs have been associated with seizure genesis because they occur in the seizure focus and during seizure onset. HFOs are also found interictally, partly co-occurring with epileptic spikes. We studied how HFOs are influenced by antiepileptic medication and seizure occurrence, to improve understanding of the pathophysiology and clinical meaning of HFOs. METHODS: Intracerebral depth EEG was partly sampled at 2,000 Hz in 42 patients with intractable focal epilepsy. Patients with five or more usable nights of recording were selected. A sample of slow-wave sleep from each night was analyzed, and HFOs (ripples: 80-250 Hz, fast ripples: 250-500 Hz) and spikes were identified on all artifact-free channels. The HFOs and spikes were compared before and after seizures with stable medication dose and during medication reduction with no intervening seizures. RESULTS: Twelve patients with five to eight nights were included. After seizures, there was an increase in spikes, whereas HFO rates remained the same. Medication reduction was followed by an increase in HFO rates and mean duration. CONCLUSIONS: Contrary to spikes, high frequency oscillations (HFOs) do not increase after seizures, but do so after medication reduction, similarly to seizures. This implies that spikes and HFOs have different pathophysiologic mechanisms and that HFOs are more tightly linked to seizures than spikes. HFOs seem to play an important role in seizure genesis and can be a useful clinical marker for disease activity. PMID- 19289738 TI - Suicidality, depression screening, and antiepileptic drugs: reaction to the FDA alert. AB - OBJECTIVE: To determine the reaction of neurology practitioners to the Food and Drug Administration (FDA) alert concerning suicidality (suicidal ideation or behavior) and antiepileptic drugs. METHODS: We designed a 21-question survey asking about the participants' approach to suicidality and depression in patients with epilepsy (PWE), and their reaction to the FDA alert and its impact on their clinical practices. Participants (n = 780) were invited via e-mail to respond to a Zoomerang survey. Two reminders were sent to increase the response. RESULTS: The survey was completed by 175 participants (22%). Most were epilepsy specialists practicing in academic settings. Almost 62% did not use any scale to routinely screen for depression in PWE. For those who used a scale, the Beck Depression Inventory was the most used one. About 42% did not feel comfortable initiating treatment for depression. Although 98% warn about behavioral side effects when starting antiepileptic drugs, only 44% warn specifically about suicidal ideations or behavior. More than half were not aware of patients who attempted to commit suicide or who had committed suicide. The mean scores for the FDA alert clarity, appropriateness, and impact on clinical practice (on a scale from 1 to 10) were low, at 5.3, 4.1, and 3.6. Almost 46% did not feel the alert is going to change their practice. CONCLUSION: The Food and Drug Administration alert did not get a favorable score from the surveyed responders. Participants actively alert patients about behavioral side effects of antiepileptic drugs, but are not specific about suicide. PMID- 19289739 TI - HIV DNA and cognition in a Thai longitudinal HAART initiation cohort: the SEARCH 001 Cohort Study. AB - OBJECTIVES: The extent to which highly active antiretroviral therapy (HAART) era cognitive disorders are due to active processes, incomplete clearance of reservoirs, or comorbidities is controversial. This study aimed to determine if immunologic and virologic factors influence cognition after first-time HAART in Thai individuals with HIV-associated dementia (HAD) and Thai individuals without HAD (non-HAD). METHODS: Variables were captured longitudinally to determine factors predictive of degree of cognitive recovery after first-time HAART. Neuropsychological data were compared to those of 230 HIV-negative Thai controls. RESULTS: HIV RNA and CD4 lymphocyte counts were not predictive of HAD cross sectionally or degree of cognitive improvement longitudinally. In contrast, baseline and longitudinal HIV DNA isolated from monocytes correlated to cognitive performance irrespective of plasma HIV RNA and CD4 lymphocyte counts pre-HAART (p < 0.001) and at 48 weeks post HAART (p < 0.001). Levels exceeding 3.5 log(10) copies HIV DNA/10(6) monocyte at baseline distinguished all HAD and non-HAD cases (p < 0.001). At 48 weeks, monocyte HIV DNA was below the level of detection of our assay (10 copies/10(6) cells) in 15/15 non-HAD compared to only 4/12 HAD cases, despite undetectable plasma HIV RNA in 26/27 cases. Baseline monocyte HIV DNA predicted 48-week cognitive performance on a composite score, independently of concurrent monocyte HIV DNA and CD4 count (p < 0.001). CONCLUSIONS: Monocyte HIV DNA level correlates to cognitive performance before highly active antiretroviral therapy (HAART) and 48 weeks after HAART in this cohort and baseline monocyte HIV DNA may predict 48-week cognitive performance. These findings raise the possibility that short-term incomplete cognitive recovery with HAART may represent an active process related to this peripheral reservoir. PMID- 19289740 TI - Hippocampal atrophy rates in Alzheimer disease: added value over whole brain volume measures. AB - OBJECTIVE: To investigate the added value of hippocampal atrophy rates over whole brain volume measurements on MRI in patients with Alzheimer disease (AD), patients with mild cognitive impairment (MCI), and controls. METHODS: We included 64 patients with AD (67 +/- 9 years; F/M 38/26), 44 patients with MCI (71 +/- 6 years; 21/23), and 34 controls (67 +/- 9 years; 16/18). Two MR scans were performed (scan interval: 1.8 +/- 0.7 years; 1.0 T), using a coronal three dimensional T1-weighted gradient echo sequence. At follow-up, 3 controls and 23 patients with MCI had progressed to AD. Hippocampi were manually delineated at baseline. Hippocampal atrophy rates were calculated using regional, nonlinear fluid registration. Whole brain baseline volumes and atrophy rates were determined using automated segmentation and registration tools. RESULTS: All MRI measures differed between groups (p < 0.005). For the distinction of MCI from controls, larger effect sizes of hippocampal measures were found compared to whole brain measures. Between MCI and AD, only whole brain atrophy rate differed significantly. Cox proportional hazards models (variables dichotomized by median) showed that within all patients without dementia, hippocampal baseline volume (hazard ratio [HR]: 5.7 [95% confidence interval: 1.5-22.2]), hippocampal atrophy rate (5.2 [1.9-14.3]), and whole brain atrophy rate (2.8 [1.1-7.2]) independently predicted progression to AD; the combination of low hippocampal volume and high atrophy rate yielded a HR of 61.1 (6.1-606.8). Within patients with MCI, only hippocampal baseline volume and atrophy rate predicted progression. CONCLUSION: Hippocampal measures, especially hippocampal atrophy rate, best discriminate mild cognitive impairment (MCI) from controls. Whole brain atrophy rate discriminates Alzheimer disease (AD) from MCI. Regional measures of hippocampal atrophy are the strongest predictors of progression to AD. PMID- 19289741 TI - Multiple sclerosis therapeutics: unexpected outcomes clouding undisputed successes. AB - In this essay, we draw attention to some recent downsides and surprises of multiple sclerosis (MS) therapeutics. These include experiences with recent head to-head trials of interferon-beta and glatiramer acetate, dose escalation trials, frustrating efforts with progressive MS trials, failures of smart concepts and designer therapies, and harsh lessons from newly observed adverse reactions. PMID- 19289742 TI - Neuropeptide Y: its multiple effects in the CNS and potential clinical significance. PMID- 19289743 TI - Sarcoidosis presenting as brainstem ischemic stroke. PMID- 19289744 TI - Hemorrhaging focal encephalitis under fingolimod (FTY720) treatment: a case report. PMID- 19289745 TI - Serum VEGF levels in POEMS syndrome and in immune-mediated neuropathies. PMID- 19289746 TI - Postinfectious ocular flutter. PMID- 19289747 TI - The varicella zoster virus vasculopathies: clinical, CSF, imaging, and virologic features. PMID- 19289748 TI - Neuropathology of brain death in the modern transplant era. PMID- 19289749 TI - Central obesity and increased risk of dementia more than three decades later. PMID- 19289750 TI - Successful treatment of eccrine angiomatous hamartoma with botulinum toxin. PMID- 19289751 TI - Lupus erythematosus tumidus: response to antimalarial treatment in 36 patients with emphasis on smoking. AB - OBJECTIVE: To determine the efficacy of antimalarial drug use in patients with lupus erythematosus tumidus. DESIGN: Retrospective single-center study. SETTING: Dermatologic clinic at a university hospital. PATIENTS: Thirty-six patients with multifocal lupus erythematosus tumidus. Intervention Treatment with either chloroquine phosphate or hydroxychloroquine sulfate. MAIN OUTCOME MEASURES: Cutaneous Lupus Erythematosus Disease Area and Severity Index score. RESULTS: Treatment with antimalarial drugs resulted in a significant reduction in the Cutaneous Lupus Erythematosus Disease Area and Severity Index score, from 4 (range, 2-8) at baseline to 1 (range, 0-6) after 3 months of therapy (P < .001). Twenty-two patients (61%) exhibited complete or almost complete clearance of skin lesions, consistent with a clinical score of 0 or 1. No difference in efficacy was noted between the chloroquine-treated group and the hydroxychloroquine treated group (P = .40). Adverse effects (nausea, dizziness, and headache) occurred only in patients treated with chloroquine. Twenty-eight patients (78%) were smokers, and smokers had a significantly higher mean (SD) clinical score than nonsmokers (5.1 [1.8] vs 3.3 [1.6]; P = .03). Moreover, smokers had a significantly lower reduction in clinical score with antimalarial treatment compared with nonsmokers (r = 0.30; P = .03; 95% confidence interval, -0.05 to 0.57). Eighty-eight percent of nonsmokers (7 of 8 patients) but only 57% of smokers (16 of 28 patients) had a clinical score of 1 or 0 after 3 months of treatment with antimalarial drugs. CONCLUSIONS: These retrospective study findings demonstrate that antimalarial treatment is highly effective in multifocal lupus erythematosus tumidus. Lower incidence of adverse effects and equal efficacy might favor the use of hydroxychloroquine. Patients who smoke should be encouraged to join smoking cessation programs because they will respond better to antimalarial treatment. PMID- 19289752 TI - Incidence of cutaneous lupus erythematosus, 1965-2005: a population-based study. AB - OBJECTIVES: To assess trends in the cutaneous variants of lupus erythematosus (CLE) and to ascertain the incidence of CLE over the past 4 decades. DESIGN: Retrospective population-based study. SETTING: Community-based epidemiology project. PATIENTS: All Olmsted County, Minnesota, residents with any subtype of CLE between January 1965 and December 2005. MAIN OUTCOME MEASURES: Incidence of CLE and disease progression to systemic LE (SLE). RESULTS: A total of 156 patients with newly diagnosed CLE (100 females and 56 males) were identified between 1965 and 2005. The incidence rate (age and sex adjusted to the 2000 US white population) was 4.30 (95% confidence interval [CI], 3.62-4.98) per 100,000. The age- and sex-adjusted prevalence as of January 1, 2006, was 73.24 (95% CI, 58.29-88.19) per 100,000. Nineteen patients with CLE had disease progression to SLE: cumulative incidence at 20 years, 19%; mean (SD) length to progression, 8.2 (6.3) years. Compared with a previously reported incidence of 2.78 (95% CI, 2.08 3.49) per 100,000 for SLE among Rochester, Minnesota, residents in 1965 through 1992, the incidence of CLE in Rochester was 3.08 (95% CI, 2.32-3.83) per 100,000 in 1965 through 1992. CONCLUSIONS: The incidence of CLE is comparable to the published incidence of SLE. Our findings double the incidence of the root designation of the disease process known as LE (SLE and CLE). PMID- 19289753 TI - Cross-sectional analysis of a collaborative Web-based database for lupus erythematosus-associated skin lesions: prospective enrollment of 114 patients. AB - OBJECTIVES: To assess disease severity in subsets of patients with cutaneous lupus erythematosus (CLE) by using outcome and quality-of-life measures, and to determine treatment responsiveness by establishing a Web-based database of patients with skin manifestations of lupus. DESIGN: Prospective, cross-sectional study. SETTING: University hospital cutaneous autoimmunity outpatient clinic. PATIENTS: One hundred fourteen patients who presented from January 15, 2007, to November 8, 2007, and met the criteria for having CLE or lupus-nonspecific skin disease. MAIN OUTCOME MEASURES: Scores on the CLE Disease Activity and Severity Index and the modified Skindex-29 (a quality-of-life measure) completed at each visit. RESULTS: Seven patients (6.1%) presented with acute CLE, 21 (18.4%) with subacute CLE, 77 (67.5%) with chronic CLE, 7 (6.1%) with systemic lupus erythematosus and LE-nonspecific skin lesions, and 1 (0.9%) with LE-nonspecific skin disease only. The mean baseline CLE Disease Activity and Severity Index activity/damage scores in patients with acute, subacute, and chronic CLE were 6.4/5.1, 11.1/1.6, and 7.5/10.2, respectively. The mean baseline modified Skindex 29 scores were 76.3, 79.4, and 82.7, respectively (P = .80). The disease in 11 of the patients (9.6%) was considered refractory to conventional therapies. Significantly more patients in the refractory group than the nonrefractory group were current smokers (P = .006). CONCLUSION: This Web-based database should allow collection of data related to disease activity, quality of life, and response to therapy at multiple centers. PMID- 19289754 TI - Prevalence of adrenal insufficiency following systemic glucocorticoid therapy in infants with hemangiomas. AB - OBJECTIVE: To determine the prevalence of adrenal insufficiency in infants with hemangiomas following treatment with systemic glucocorticoids (GCs). DESIGN: Prospective study for 18 months. SETTING: Hemangioma and vascular malformation center at a tertiary care children's hospital. PATIENTS: Sixteen infants with hemangiomas had an adrenal axis evaluation as soon as possible following the completion of GC therapy. Ten healthy control infants were also evaluated for comparison. INTERVENTIONS: Prednisolone at a starting dose of 2 to 3 mg/kg/d for 4 weeks, followed by a tapering period. The mean duration of GC treatment was 7.2 months. MAIN OUTCOME MEASURE: Prevalence of adrenal insufficiency in GC-treated subjects as assessed by a combination low-dose/high-dose corticotropin stimulation test. RESULTS: Subjects underwent corticotropin testing at a mean of 13 days after the completion of therapy. Only 1 of the 16 GC-treated infants (6%) had adrenal insufficiency. This subject was tested 1 day after GC treatment was stopped, and results from retesting 3 months later were normal. All control subjects had normal adrenal function. CONCLUSION: Infants with hemangiomas are at low risk of adrenal insufficiency following the completion of GC therapy, as used in our hemangioma center. PMID- 19289755 TI - Evaluation of the efficacy of acitretin therapy for nail psoriasis. AB - OBJECTIVE: To evaluate the therapeutic efficacy of acitretin in patients with isolated nail psoriasis. DESIGN: Open study involving 36 patients with moderate to severe nail psoriasis treated with acitretin. SETTING: University-based outpatient dermatology clinic specializing in nail diseases. PATIENTS: A total of 27 men and 9 women (mean age, 41 years) with nail psoriasis. INTERVENTION: Therapy consisted of acitretin, 0.2 to 0.3 mg/kg/d, for 6 months. MAIN OUTCOME MEASURES: Clinical evaluation, and Nail Psoriasis Severity Index (NAPSI) and modified NAPSI scores before therapy, every 2 months during therapy, and 6 months after treatment. RESULTS: The mean percentage of reduction of the NAPSI score after treatment was 41%; the mean percentage of reduction of the modified NAPSI score of the target nail was 50%. Clinical evaluation at 6 months showed complete or almost complete clearing of the nail lesions in 9 patients (25%), moderate improvement in 9 (25%), mild improvement in 12 (33%), and no improvement in 6 (11%). CONCLUSION: Results from low-dose acitretin therapy show NAPSI score reductions comparable with those studies evaluating biologic drugs for nail psoriasis and suggest that low-dose systemic acitretin should be considered in the treatment of nail psoriasis. PMID- 19289756 TI - Successful treatment of complex aphthosis with colchicine and dapsone. AB - OBJECTIVE: To investigate the effectiveness of colchicine and dapsone, 2 corticosteroid-sparing anti-inflammatory agents, in the treatment of patients with complex aphthosis (recurrent oral and genital aphthous ulcers or severe, almost constant, multiple oral aphthae in the absence of Behcet syndrome). DESIGN: Retrospective review of medical records. SETTING: Tertiary care medical clinic. PATIENTS: Fifty-five patients with complex aphthosis evaluated and treated at Mayo Clinic between January 1, 1998, and July 31, 2007. All the patients were treated according to a therapeutic ladder, starting with colchicine and adding dapsone to treatment of patients who did not have a substantial response (>75% improvement) to colchicine or who discontinued colchicine use because of adverse effects. MAIN OUTCOME MEASURES: A substantial response to therapy with colchicine alone, dapsone alone, or colchicine and dapsone combined. RESULTS: Most patients (44 [80%]) had a substantial response to therapy and had no serious adverse effects. CONCLUSIONS: Colchicine and dapsone are effective, safe therapies for the treatment of complex aphthosis. Colchicine and dapsone, 2 established drugs also used for gout and leprosy, respectively, and for other dermatologic disorders, should be considered efficacious in the treatment of complex aphthosis. PMID- 19289757 TI - Necrobiotic xanthogranuloma: a review of 17 cases with emphasis on clinical and pathologic correlation. AB - OBJECTIVE: To identify correlations between clinical presentation, specific histopathologic findings, and subsequent disease course in patients with necrobiotic xanthogranuloma (NXG). DESIGN: Retrospective review of medical records and histopathologic examination of fixed tissue samples. SETTING: Tertiary care medical center. PATIENTS: Seventeen patients with a diagnosis of NXG established between January 1, 1994, and December 31, 2007. MAIN OUTCOME MEASURES: Description and distribution of clinical lesions, presence of monoclonal gammopathy, multiple myeloma, and correlation with microscopic patterns of skin lesions. RESULTS: Eleven patients (65%) showed involvement of the periorbital area, and the trunk was affected in 8 patients (47%). Twelve patients (71%) had a monoclonal gammopathy; of these, 3 (18%) had multiple myeloma. Histopathologic examination of 12 patients showed findings consistent with NXG, including a bandlike pattern of necrobiotic granulomatous inflammation, atypical giant cells, cholesterol clefts, and plasma cells. No correlations were identified between clinical presentation and specific histopathologic findings. Although most patients had a serum monoclonal gammopathy, staining with antibodies to CD3, CD20, kappa light chains, and lambda light chains showed polytypic lymphocytes and plasma cells in all cases. CONCLUSIONS: The association between NXG and paraproteinemia is well documented and corroborated by this study. However, the skin lesions in NXG represent reactive inflammation and are not associated with the presence of monoclonal plasma cells or multiple myeloma. PMID- 19289758 TI - Mount Tsukuba and the origin of tacrolimus. PMID- 19289759 TI - The misnomer "macrocephaly-cutis marmorata telangiectatica congenita syndrome": report of 12 new cases and support for revising the name to macrocephaly capillary malformations. AB - BACKGROUND: The condition known as macrocephaly-cutis marmorata telangiectatica congenita syndrome (M-CMTC) is a rare congenital syndrome of unknown etiology characterized by macrocephaly and vascular lesions that have been described as either cutis marmorata or cutis marmorata telangiectatica congenita (CMTC). Most patients also exhibit facial and limb asymmetry; somatic overgrowth; developmental delay; capillary malformations of the nose, philtrum, and/or upper lip; neurologic abnormalities; syndactyly or polydactyly; craniofacial abnormalities; and joint laxity or soft skin. OBSERVATIONS: We describe 12 patients with this condition from tertiary care medical centers (8 cases) and accrued via an M-CMTC support group Web site (4 cases). All patients showed reticulated or confluent port-wine stains (PWS), not CMTC. Seven of the 12 patients also had centrofacial capillary malformations. In our comprehensive review of 100 previously reported cases, only 34 were accompanied by photographs that were sufficiently clear to review for diagnostic purposes. None had true CMTC, with most having reticulated PWS or persistent cutis marmorata. CONCLUSIONS: Reticulated or confluent PWS and persistent capillary malformations of the central face, rather than CMTC, are the most characteristic cutaneous vascular anomalies seen in so-called M-CMTC syndrome. The name macrocephaly capillary malformations (M-CM) more accurately reflects the features of this syndrome. PMID- 19289760 TI - Neurolymphomatosis associated with Sezary syndrome. AB - BACKGROUND: Mycosis fungoides and Sezary syndrome are cutaneous T-cell lymphomas characterized by the epidermotropism of tumor cells. Neuropathic disease is rare during mycosis fungoides and Sezary syndrome and usually results from a central nervous system involvement in late stages. Neurolymphomatosis is defined as the infiltration of the peripheral nerves by tumor lymphocytes. It has been described in patients with aggressive systemic lymphomas but, to our knowledge, not in patients with mycosis fungoides or Sezary syndrome. We report the first case of neurolymphomatosis in a patient with Sezary syndrome and the partial efficacy of high-dose methotrexate sodium in treating this usually refractory complication. OBSERVATION: A 73-year-old woman with newly diagnosed Sezary syndrome rapidly developed severe peripheral neuropathic disease with multiple paralyses. Biopsy specimens were taken from a clinically affected nerve and the adjacent muscle; they revealed a neural infiltration by Sezary cells with secondary muscular atrophy. Partial response and major neurologic recovery occurred and persisted under high doses of intravenous methotrexate until the patient died 14 months after the Sezary syndrome diagnosis from a pericarditis of uncertain origin. CONCLUSION: This unusual and demonstrative case report highlights the possible neurotropism of malignant cells in Sezary syndrome and suggests the effectiveness of high doses of intravenous methotrexate in this rare and fatal disorder. PMID- 19289761 TI - Isolated benign primary cutaneous plasmacytosis in children: two illustrative cases. AB - BACKGROUND: Plasma cells are normally found in bone marrow and the intestinal tract. They appear in the skin in malignant conditions, autoimmune diseases, infection, and idiopathic and poorly understood disorders such as primary nodular amyloidosis. It is uncommon to find collections of plasma cells in the skin in the absence of these conditions. OBSERVATIONS: We present 2 cases of cutaneous plasmacytosis, one in a white, female adolescent aged 15 years with an 11-year history of a solitary, asymptomatic, violaceous plaque on the left anterior tibia and the other in a white, male child aged 7 years with a 2-year history of a solitary erythematous plaque on the right anterior tibia. In both patients, infiltration of mature polyclonal plasma cells was confined to an area on the skin with papulonodules. There was no history of previous trauma, malignant conditions, autoimmune disease, or infection in either child. CONCLUSION: Although incipient or occult systemic disease cannot be definitively ruled out, the course of these 2 individuals suggests that isolated primary cutaneous plasmacytosis in children is a benign chronic process with no adverse sequelae. PMID- 19289762 TI - Lenalidomide for the treatment of resistant discoid lupus erythematosus. AB - BACKGROUND: Discoid lupus erythematosus (DLE) is a chronic, disfiguring disease that is characterized by scaly, erythematous, disk-shaped patches and plaques followed by atrophy, scarring, and dyspigmentation. It is refractory to standard therapies in a small population of patients. We investigated the use of lenalidomide, a thalidomide analogue, as a novel alternative therapy in 2 cases of refractory DLE and report our results. OBSERVATIONS: Two patients with chronic, severe DLE were treated with low-dose lenalidomide. Improvement was noted within 1 month at a dosage of 5 mg/d in one case and was maintained for 10 months before the dosage was doubled to 10 mg/d for 12 months because of a slight worsening of symptoms. Clinical improvement was demonstrated by a sustained reduction in the Cutaneous Lupus Erythematosus Disease Area and Severity Index activity score, with no change in the Cutaneous Lupus Erythematosus Disease Area and Severity Index damage score. Within 5 months, oral prednisone therapy (60 mg/d) was tapered and discontinued; it was restarted at a low dosage (5 mg/d), however, to manage the symptoms of systemic LE. Of note, the patient experienced mild neutropenia after taking 10 mg/d of lenalidomide, which carries a black box warning regarding neutropenia; therefore, the complete blood cell count should be monitored weekly for the first 2 months and then monthly thereafter. The second case failed to show clinical improvement, and lenalidomide therapy was discontinued after 6 months. CONCLUSIONS: Lenalidomide therapy is a potential alternative or adjunctive treatment for patients with severe, chronic DLE that is refractory to standard therapies. A larger study is needed to clarify its role in the treatment of DLE and other forms of cutaneous LE. PMID- 19289763 TI - Melanoma: do we need a hatchet or a scalpel? PMID- 19289764 TI - Accuracy of diagnosis of pediculosis capitis: visual inspection vs wet combing. AB - OBJECTIVE: To determine the diagnostic accuracy of visual inspection and wet combing in pediculosis capitis (head lice infestation). Visual inspection of 5 predilection sites (temples, behind the ears, and neck) was performed first, followed by wet combing of hair moistened with conditioner. Presence of mobile stages was defined as active infestation, presence of nits alone as historic infestation. DESIGN: Observer-blinded comparison of 2 diagnostic methods. SETTING: Five primary schools in which head lice infestation was epidemic. PARTICIPANTS: A total of 304 students aged 6 to 12 years. MAIN OUTCOME MEASURES: Presence of nymph, adults, and nits; sensitivity, predictive value, and accuracy of both methods. RESULTS: Visual inspection underestimated the true prevalence of active infestation by a factor of 3.5. The sensitivity of wet combing in diagnosing active infestation was significantly higher than of visual inspection (90.5% vs 28.6%; P < .001). The accuracy of the former method was 99.3% and that of the latter method, 95%. In contrast, visual inspection had a higher sensitivity for the diagnosis of historic infestation (86.1% vs 68.4%; P < .001). CONCLUSIONS: Wet combing is a very accurate method to diagnose active head lice infestation. Visual inspection is the method of choice, if one aims to determine the frequency of carriers of eggs or nits. PMID- 19289765 TI - Alitretinoin as a potential advance in the management of severe chronic hand eczema. PMID- 19289766 TI - Clinically relevant information about cutaneous lupus erythematosus. PMID- 19289767 TI - Hypothalamic-pituitary-adrenal axis suppression in systemic glucocorticoid treated infantile hemangiomas: putting the risk into context. PMID- 19289768 TI - Atrophic scars in a 54-year-old woman. PMID- 19289769 TI - Multiple violaceous nodules in a neonate. PMID- 19289770 TI - A solitary nodule on the finger. PMID- 19289771 TI - Pruritic papules limited to tattooed skin. PMID- 19289772 TI - Treatment of cutaneous B-cell lymphoma, leg type, with age-adapted combinations of chemotherapies and rituximab. PMID- 19289773 TI - Ultrasound therapy for lipodermatosclerosis. PMID- 19289774 TI - Use of photographs illustrating ABCDE criteria in skin self-examination. PMID- 19289775 TI - Lichen planopilaris: retrospective study and stepwise therapeutic approach. PMID- 19289776 TI - Intravenous immunoglobulin therapy for dystrophic calcinosis cutis: unreliable in our hands. PMID- 19289777 TI - Mentoring satisfied residents. PMID- 19289778 TI - A call for more dermatologic input into chronic graft-vs-host disease clinical trials. PMID- 19289779 TI - The importance of caution in the use of unregulated anticellulite treatments. PMID- 19289780 TI - Roosevelt's pigmented lesion: probably not a melanoma. PMID- 19289781 TI - Subacute cutaneous lupus erythematosus associated with capecitabine monotherapy. PMID- 19289782 TI - Autoimmune progesterone dermatitis. PMID- 19289783 TI - Atypical linear IgA dermatosis revealing angioimmunoblastic T-cell lymphoma. PMID- 19289784 TI - Tumid lupus erythematosus at the site of a scar. PMID- 19289785 TI - Sweet syndrome with pulmonary involvement in a healthy young woman. PMID- 19289786 TI - Stage III melanoma treated with chemotherapy after surgery during the second trimester of pregnancy. PMID- 19289787 TI - Tunga penetrans: description of a new dermoscopic sign--the radial crown. PMID- 19289788 TI - Type 1 leprosy reaction manifesting after discontinuation of adalimumab therapy. PMID- 19289789 TI - Ignatius J. Reilly's flamboyant dermatosis. PMID- 19289790 TI - Dermoscopy in active discoid lupus. PMID- 19289791 TI - GSK3beta phosphorylation modulates CLASP-microtubule association and lamella microtubule attachment. AB - Polarity of the microtubule (MT) cytoskeleton is essential for many cell functions. Cytoplasmic linker-associated proteins (CLASPs) are MT-associated proteins thought to organize intracellular MTs and display a unique spatiotemporal regulation. In migrating epithelial cells, CLASPs track MT plus ends in the cell body but bind along MTs in the lamella. In this study, we demonstrate that glycogen synthase kinase 3beta (GSK3beta) directly phosphorylates CLASPs at multiple sites in the domain required for MT plus end tracking. Although complete phosphorylation disrupts both plus end tracking and association along lamella MTs, we show that partial phosphorylation of the identified GSK3beta motifs determines whether CLASPs track plus ends or associate along MTs. In addition, we find that expression of constitutively active GSK3beta destabilizes lamella MTs by disrupting lateral MT interactions with the cell cortex. GSK3beta-induced lamella MT destabilization was partially rescued by expression of CLASP2 with mutated phosphorylation sites. This indicates that CLASP-mediated stabilization of peripheral MTs, which likely occurs in the vicinity of focal adhesions, may be regulated by local GSK3beta inactivation. PMID- 19289792 TI - Wac: a new Augmin subunit required for chromosome alignment but not for acentrosomal microtubule assembly in female meiosis. AB - The bipolar spindle forms without centrosomes naturally in female meiosis and by experimental manipulation in mitosis. Augmin is a recently discovered protein complex required for centrosome-independent microtubule generation within the spindle in Drosophila melanogaster cultured cells. Five subunits of Augmin have been identified so far, but neither their organization within the complex nor their role in developing organisms is known. In this study, we report a new Augmin subunit, wee Augmin component (Wac). Wac directly interacts with another Augmin subunit, Dgt2, via its coiled-coil domain. Wac depletion in cultured cells, especially without functional centrosomes, causes severe defects in spindle assembly. We found that a wac deletion mutant is viable but female sterile and shows only a mild impact on somatic mitosis. Unexpectedly, mutant female meiosis showed robust microtubule assembly of the acentrosomal spindle but frequent chromosome misalignment. For the first time, this study establishes the role of an Augmin subunit in developing organisms and provides an insight into the architecture of the complex. PMID- 19289793 TI - Sem1 is a functional component of the nuclear pore complex-associated messenger RNA export machinery. AB - The evolutionarily conserved protein Sem1/Dss1 is a subunit of the regulatory particle (RP) of the proteasome, and, in mammalian cells, binds the tumor suppressor protein BRCA2. Here, we describe a new function for yeast Sem1. We show that sem1 mutants are impaired in messenger RNA (mRNA) export and transcription elongation, and induce strong transcription-associated hyper recombination phenotypes. Importantly, Sem1, independent of the RP, is functionally linked to the mRNA export pathway. Biochemical analyses revealed that, in addition to the RP, Sem1 coenriches with components of two other multisubunit complexes: the nuclear pore complex (NPC)-associated TREX-2 complex that is required for transcription-coupled mRNA export, and the COP9 signalosome, which is involved in deneddylation. Notably, targeting of Thp1, a TREX-2 component, to the NPC is perturbed in a sem1 mutant. These findings reveal an unexpected nonproteasomal function of Sem1 in mRNA export and in prevention of transcription-associated genome instability. Thus, Sem1 is a versatile protein that might stabilize multiple protein complexes involved in diverse pathways. PMID- 19289794 TI - A clathrin-dependent pathway leads to KRas signaling on late endosomes en route to lysosomes. AB - Ras proteins are small guanosine triphosphatases involved in the regulation of important cellular functions such as proliferation, differentiation, and apoptosis. Understanding the intracellular trafficking of Ras proteins is crucial to identify novel Ras signaling platforms. In this study, we report that epidermal growth factor triggers Kirsten Ras (KRas) translocation onto endosomal membranes (independently of calmodulin and protein kinase C phosphorylation) through a clathrin-dependent pathway. From early endosomes, KRas but not Harvey Ras or neuroblastoma Ras is sorted and transported to late endosomes (LEs) and lysosomes. Using yellow fluorescent protein-Raf1 and the Raichu-KRas probe, we identified for the first time in vivo-active KRas on Rab7 LEs, eliciting a signal output through Raf1. On these LEs, we also identified the p14-MP1 scaffolding complex and activated extracellular signal-regulated kinase 1/2. Abrogation of lysosomal function leads to a sustained late endosomal mitogen-activated protein kinase signal output. Altogether, this study reveals novel aspects about KRas intracellular trafficking and signaling, shedding new light on the mechanisms controlling Ras regulation in the cell. PMID- 19289795 TI - TopBP1 and DNA polymerase-alpha directly recruit the 9-1-1 complex to stalled DNA replication forks. AB - TopBP1 and the Rad9-Rad1-Hus1 (9-1-1) complex activate the ataxia telangiectasia mutated and Rad3-related (ATR) protein kinase at stalled replication forks. ATR is recruited to stalled forks through its binding partner, ATR-interacting protein (ATRIP); however, it is unclear how TopBP1 and 9-1-1 are recruited so that they may join ATR-ATRIP and initiate signaling. In this study, we use Xenopus laevis egg extracts to determine the requirements for 9-1-1 loading. We show that TopBP1 is required for the recruitment of both 9-1-1 and DNA polymerase (pol)-alpha to sites of replication stress. Furthermore, we show that pol-alpha is also directly required for Rad9 loading. Our study identifies an assembly pathway, which is controlled by TopBP1 and includes pol-alpha, that mediates the loading of the 9-1-1 complex onto stalled replication forks. These findings clarify early events in the assembly of checkpoint signaling complexes on DNA and identify TopBP1 as a critical sensor of replication stress. PMID- 19289796 TI - In search of nonribosomal nucleolar protein function and regulation. AB - The life of the nucleolus has proven to be more colorful and multifaceted than had been envisioned a decade ago. A large number of proteins found in this subnuclear compartment have no identifiable tie either to the ribosome biosynthetic pathway or to the other newly established activities occurring within the nucleolus. The questions of how and why these proteins end up in this subnuclear compartment remain unanswered and are the focus of intense current interest. This review discusses our thoughts on the discovery of nonribosomal proteins in the nucleolus. PMID- 19289798 TI - Research ethics review: do it once and do it well. PMID- 19289799 TI - Effective and efficient use of implantable defibrillators: sometimes it's over when it's over. PMID- 19289800 TI - Is diagnosing metabolic syndrome a uniquely simple way to predict incident type 2 diabetes mellitus? PMID- 19289801 TI - Clinical trials: the balance between protecting participants and promoting drug and product development. PMID- 19289802 TI - HIV scandal sinks Chilean health minister. PMID- 19289803 TI - On the ground in the Gaza Strip. PMID- 19289804 TI - Maximum potential benefit of implantable defibrillators in preventing sudden death after hospital admission because of heart failure. AB - BACKGROUND: Implantable defibrillators are recommended for the prevention of sudden cardiac death in patients with heart failure. However, criteria to identify those who would benefit most from this therapy are lacking. We assessed the maximum potential benefit of preventing sudden death in patients with repeated hospital admissions because of heart failure. METHODS: Using a cohort assembled from an administrative database, we identified 14,374 patients admitted to hospital for the first time because of heart failure between Jan. 1, 2000, and Dec. 31, 2004. We followed subsequent admissions related to heart failure as well as mortality and causes of death to Mar. 31, 2006. We regarded all out-of hospital cardiac deaths as sudden deaths. We calculated the maximum potential benefit of preventing sudden death by subtracting the observed survival after each hospital admission from the hypothetical survival whereby all out-of hospital cardiac deaths were assumed to be preventable. RESULTS: The mean age of the cohort was 77 years, 45% were women, 11% had cerebrovascular disease, and 21% had chronic kidney disease. Out-of-hospital cardiac deaths constituted 13.7% (1226/8967) of all deaths during 32,055 person-years of follow-up. The median survival declined with each subsequent hospital admission related to heart failure. The hypothetical prevention of all out-of-hospital deaths prolonged life by 0.63 (95% confidence interval [CI] 0.49 to 0.77) years after the first hospital admission. This potential benefit dropped to 0.28 (95% CI 0.10 to 0.46) years after 3 hospital admissions related to heart failure. Among patients less than 65 years old, and older patients without kidney disease, dementia or cancer, more than 50% survived longer than 2 years until they had 2 or 3 hospital admissions related to heart failure. INTERPRETATION: The use of implantable defibrillators to prevent sudden death would provide limited benefit among older patients with comorbidities and among patients with multiple hospital admissions related to heart failure. PMID- 19289805 TI - Metabolic syndrome and its components as predictors of incident type 2 diabetes mellitus in an Aboriginal community. AB - BACKGROUND: Risk factors for type 2 diabetes remain poorly characterized among Aboriginal Canadians. We aimed to determine the incidence of type 2 diabetes in an Aboriginal community and to evaluate prospective associations with metabolic syndrome and its components. METHODS: Of 606 participants in the Sandy Lake Health and Diabetes Project from 1993 to 1995 who were free of diabetes at baseline, 540 (89.1%) participated in 10-year follow-up assessments. Baseline anthropometry, blood pressure, fasting insulin and serum lipid levels were measured. Fasting and 2-hour postload glucose levels were obtained at follow-up to determine incident cases of type 2 diabetes. RESULTS: The 10-year cumulative incidence of diabetes was 17.5%. High adiposity, dyslipidemia, hyperglycemia, hyperinsulinemia and hypertension at baseline were associated with an increased risk of diabetes after adjustment for age and sex (all p < or = 0.03). Metabolic syndrome had high specificity (75%-88%) and high negative predictive value (85% 87%) to correctly detect diabetes-free individuals at follow-up. It had low sensitivity (26%-48%) and low positive predictive value (29%-32%) to detect future diabetes. Metabolic syndrome at baseline was associated with incident diabetes after adjustment for age and sex, regardless of whether the syndrome was defined using the National Cholesterol Education Program criteria (odds ratio [OR] 2.03, 95% confidence interval [CI] 1.10-3.75) or the International Diabetes Federation criteria (OR 2.14, 95% CI 1.29-3.55). The association was to the same degree as that for impaired glucose tolerance assessed using the oral glucose tolerance test (OR 2.87, 95% CI 1.52-5.40; p > 0.05 for comparison of C statistics). INTERPRETATION: Metabolic syndrome and its components can be identified with readily available clinical measures. As such, the syndrome may be useful for identifying individuals at risk of type 2 diabetes in remote Aboriginal communities. PMID- 19289806 TI - Generalized tonic-clonic seizure after a taser shot to the head. AB - During a police chase on foot, a previously well police officer was hit mistakenly by a taser shot meant for the suspect. The taser gun had been fired once, sending 2 barbed darts into his upper back and occiput. Within seconds, the officer collapsed and experienced a generalized tonic-clonic seizure with loss of consciousness and postictal confusion. Subsequent magnetic resonance imaging scans of the head and electroencephalograms were normal. The patient has experienced no recurrence of seizure over more than a year of follow-up. This report shows that a taser shot to the head may result in a brain-specific complication such as generalized tonic-clonic seizure. It also suggests that seizure should be considered an adverse event related to taser use. PMID- 19289807 TI - Hospital infection control strategies for vancomycin-resistant Enterococcus, methicillin-resistant Staphylococcus aureus and Clostridium difficile. PMID- 19289808 TI - Are dietary recommendations for the use of fish oils sustainable? PMID- 19289809 TI - Neurocysticercosis in a patient in Canada. PMID- 19289811 TI - Proton pump inhibitors and osteoporosis-related fractures. PMID- 19289812 TI - Apology laws. PMID- 19289813 TI - In the face of cancer. PMID- 19289814 TI - Here be dragons? PMID- 19289816 TI - Half-and-half nail. PMID- 19289817 TI - Criminalizing HIV transmission may be a mistake. PMID- 19289818 TI - Medial temporal theta state before an event predicts episodic encoding success in humans. AB - We report a human electrophysiological brain state that predicts successful memory for events before they occur. Using magnetoencephalographic recordings of brain activity during episodic memory encoding, we show that amplitudes of theta oscillations shortly preceding the onsets of words were higher for later-recalled than for later-forgotten words. Furthermore, single-trial analyses revealed that recall rate in all 24 participants tested increased as a function of increasing prestimulus theta amplitude. This positive correlation was independent of whether participants were preparing for semantic or phonemic stimulus processing, thus likely signifying a memory-related theta state rather than a preparatory task set. Source analysis located this theta state to the medial temporal lobe, a region known to be critical for encoding and recall. These findings provide insight into state-related aspects of memory formation in humans, and open a perspective for improving memory through theta-related brain states. PMID- 19289819 TI - Prostaglandin mediates IL-23/IL-17-induced neutrophil migration in inflammation by inhibiting IL-12 and IFNgamma production. AB - IL-23/IL-17-induced neutrophil recruitment plays a pivotal role in rheumatoid arthritis (RA). However, the mechanism of the neutrophil recruitment is obscure. Here we report that prostaglandin enhances the IL-23/IL-17-induced neutrophil migration in a murine model of RA by inhibiting IL-12 and IFN gamma production. Methylated BSA (mBSA) and IL-23-induced neutrophil migration was inhibited by anti-IL-23 and anti-IL-17 antibodies, COX inhibitors, IL-12, or IFNgamma but was enhanced by prostaglandin E(2) (PGE(2)). IL-23-induced IL-17 production was increased by PGE(2) and suppressed by COX-inhibition or IL-12. Furthermore, COX inhibition failed to reduce IL-23-induced neutrophil migration in IL-12- or IFNgamma-deficient mice. IL-17-induced neutrophil migration was not affected by COX inhibitors, IL-12, or IFNgamma but was inhibited by MK886 (a leukotriene synthesis inhibitor), anti-TNFalpha, anti-CXCL1, and anti-CXCL5 antibodies and by repertaxin (a CXCR1/2 antagonist). These treatments all inhibited mBSA- or IL-23 induced neutrophil migration. IL-17 induced neutrophil chemotaxis through a CXC chemokines-dependent pathway. Our results suggest that prostaglandin plays an important role in IL-23-induced neutrophil migration in arthritis by enhancing IL 17 synthesis and by inhibiting IL-12 and IFNgamma production. We thus provide a mechanism for the pathogenic role of the IL-23/IL-17 axis in RA and also suggest an additional mechanism of action for nonsteroidal anti-inflammatory drugs. PMID- 19289820 TI - Solution to the problem of the poor cyclic fatigue resistance of bulk metallic glasses. AB - The recent development of metallic glass-matrix composites represents a particular milestone in engineering materials for structural applications owing to their remarkable combination of strength and toughness. However, metallic glasses are highly susceptible to cyclic fatigue damage, and previous attempts to solve this problem have been largely disappointing. Here, we propose and demonstrate a microstructural design strategy to overcome this limitation by matching the microstructural length scales (of the second phase) to mechanical crack-length scales. Specifically, semisolid processing is used to optimize the volume fraction, morphology, and size of second-phase dendrites to confine any initial deformation (shear banding) to the glassy regions separating dendrite arms having length scales of approximately 2 mum, i.e., to less than the critical crack size for failure. Confinement of the damage to such interdendritic regions results in enhancement of fatigue lifetimes and increases the fatigue limit by an order of magnitude, making these "designed" composites as resistant to fatigue damage as high-strength steels and aluminum alloys. These design strategies can be universally applied to any other metallic glass systems. PMID- 19289821 TI - Characterization of Drosophila melanogaster cytochrome P450 genes. AB - Cytochrome P450s form a large and diverse family of heme-containing proteins capable of carrying out many different enzymatic reactions. In both mammals and plants, some P450s are known to carry out reactions essential for processes such as hormone synthesis, while other P450s are involved in the detoxification of environmental compounds. In general, functions of insect P450s are less well understood. We characterized Drosophila melanogaster P450 expression patterns in embryos and 2 stages of third instar larvae. We identified numerous P450s expressed in the fat body, Malpighian (renal) tubules, and in distinct regions of the midgut, consistent with hypothesized roles in detoxification processes, and other P450s expressed in organs such as the gonads, corpora allata, oenocytes, hindgut, and brain. Combining expression pattern data with an RNA interference lethality screen of individual P450s, we identify candidate P450s essential for developmental processes and distinguish them from P450s with potential functions in detoxification. PMID- 19289822 TI - miR-375 maintains normal pancreatic alpha- and beta-cell mass. AB - Altered growth and development of the endocrine pancreas is a frequent cause of the hyperglycemia associated with diabetes. Here we show that microRNA-375 (miR 375), which is highly expressed in pancreatic islets, is required for normal glucose homeostasis. Mice lacking miR-375 (375KO) are hyperglycemic, exhibit increased total pancreatic alpha-cell numbers, fasting and fed plasma glucagon levels, and increased gluconeogenesis and hepatic glucose output. Furthermore, pancreatic beta-cell mass is decreased in 375KO mice as a result of impaired proliferation. In contrast, pancreatic islets of obese mice (ob/ob), a model of increased beta-cell mass, exhibit increased expression of miR-375. Genetic deletion of miR-375 from these animals (375/ob) profoundly diminished the proliferative capacity of the endocrine pancreas and resulted in a severely diabetic state. Bioinformatic analysis of transcript data from 375KO islets revealed that miR-375 regulates a cluster of genes controlling cellular growth and proliferation. These data provide evidence that miR-375 is essential for normal glucose homeostasis, alpha- and beta-cell turnover, and adaptive beta-cell expansion in response to increasing insulin demand in insulin resistance. PMID- 19289823 TI - Seizures, sensorineural deafness, ataxia, mental retardation, and electrolyte imbalance (SeSAME syndrome) caused by mutations in KCNJ10. AB - We describe members of 4 kindreds with a previously unrecognized syndrome characterized by seizures, sensorineural deafness, ataxia, mental retardation, and electrolyte imbalance (hypokalemia, metabolic alkalosis, and hypomagnesemia). By analysis of linkage we localize the putative causative gene to a 2.5-Mb segment of chromosome 1q23.2-23.3. Direct DNA sequencing of KCNJ10, which encodes an inwardly rectifying K(+) channel, identifies previously unidentified missense or nonsense mutations on both alleles in all affected subjects. These mutations alter highly conserved amino acids and are absent among control chromosomes. Many of these mutations have been shown to cause loss of function in related K(+) channels. These findings demonstrate that loss-of-function mutations in KCNJ10 cause this syndrome, which we name SeSAME. KCNJ10 is expressed in glia in the brain and spinal cord, where it is believed to take up K(+) released by neuronal repolarization, in cochlea, where it is involved in the generation of endolymph, and on the basolateral membrane in the distal nephron. We propose that KCNJ10 is required in the kidney for normal salt reabsorption in the distal convoluted tubule because of the need for K(+) recycling across the basolateral membrane to enable normal activity of the Na(+)-K(+)-ATPase; loss of this function accounts for the observed electrolyte defects. Mice deficient for KCNJ10 show a related phenotype with seizures, ataxia, and hearing loss, further supporting KCNJ10's role in this syndrome. These findings define a unique human syndrome, and establish the essential role of basolateral K(+) channels in renal electrolyte homeostasis. PMID- 19289824 TI - Thinking like a trader selectively reduces individuals' loss aversion. AB - Research on emotion regulation has focused upon observers' ability to regulate their emotional reaction to stimuli such as affective pictures, but many other aspects of our affective experience are also potentially amenable to intentional cognitive regulation. In the domain of decision-making, recent work has demonstrated a role for emotions in choice, although such work has generally remained agnostic about the specific role of emotion. Combining psychologically derived cognitive strategies, physiological measurements of arousal, and an economic model of behavior, this study examined changes in choices (specifically, loss aversion) and physiological correlates of behavior as the result of an intentional cognitive regulation strategy. Participants were on average more aroused per dollar to losses relative to gains, as measured with skin conductance response, and the difference in arousal to losses versus gains correlated with behavioral loss aversion across subjects. These results suggest a specific role for arousal responses in loss aversion. Most importantly, the intentional cognitive regulation strategy, which emphasized "perspective-taking," uniquely reduced both behavioral loss aversion and arousal to losses relative to gains, largely by influencing arousal to losses. Our results confirm previous research demonstrating loss aversion while providing new evidence characterizing individual differences and arousal correlates and illustrating the effectiveness of intentional regulation strategies in reducing loss aversion both behaviorally and physiologically. PMID- 19289825 TI - An adrenal beta-arrestin 1-mediated signaling pathway underlies angiotensin II induced aldosterone production in vitro and in vivo. AB - Aldosterone produces a multitude of effects in vivo, including promotion of postmyocardial infarction adverse cardiac remodeling and heart failure progression. It is produced and secreted by the adrenocortical zona glomerulosa (AZG) cells after angiotensin II (AngII) activation of AngII type 1 receptors (AT(1)Rs). Until now, the general consensus for AngII signaling to aldosterone production has been that it proceeds via activation of G(q/11)-proteins, to which the AT(1)R normally couples. Here, we describe a novel signaling pathway underlying this AT(1)R-dependent aldosterone production mediated by beta-arrestin 1 (betaarr1), a universal heptahelical receptor adapter/scaffolding protein. This pathway results in sustained ERK activation and subsequent up-regulation of steroidogenic acute regulatory protein, a steroid transport protein regulating aldosterone biosynthesis in AZG cells. Also, this betaarr1-mediated pathway appears capable of promoting aldosterone turnover independently of G protein activation, because treatment of AZG cells with SII, an AngII analog that induces betaarr, but not G protein coupling to the AT(1)R, recapitulates the effects of AngII on aldosterone production and secretion. In vivo, increased adrenal betaarr1 activity, by means of adrenal-targeted adenoviral-mediated gene delivery of a betaarr1 transgene, resulted in a marked elevation of circulating aldosterone levels in otherwise normal animals, suggesting that this adrenocortical betaarr1-mediated signaling pathway is operative, and promotes aldosterone production and secretion in vivo, as well. Thus, inhibition of adrenal betaarr1 activity on AT(1)Rs might be of therapeutic value in pathological conditions characterized and aggravated by hyperaldosteronism. PMID- 19289826 TI - Enrichment of the amino acid L-isovaline by aqueous alteration on CI and CM meteorite parent bodies. AB - The distribution and enantiomeric composition of the 5-carbon (C(5)) amino acids found in CI-, CM-, and CR-type carbonaceous meteorites were investigated by using liquid chromatography fluorescence detection/TOF-MS coupled with o phthaldialdehyde/N-acetyl-L-cysteine derivatization. A large L-enantiomeric excess (ee) of the alpha-methyl amino acid isovaline was found in the CM meteorite Murchison (L(ee) = 18.5 +/- 2.6%) and the CI meteorite Orgueil (L(ee) = 15.2 +/- 4.0%). The measured value for Murchison is the largest enantiomeric excess in any meteorite reported to date, and the Orgueil measurement of an isovaline excess has not been reported previously for this or any CI meteorite. The L-isovaline enrichments in these two carbonaceous meteorites cannot be the result of interference from other C(5) amino acid isomers present in the samples, analytical biases, or terrestrial amino acid contamination. We observed no L isovaline enrichment for the most primitive unaltered Antarctic CR meteorites EET 92042 and QUE 99177. These results are inconsistent with UV circularly polarized light as the primary mechanism for L-isovaline enrichment and indicate that amplification of a small initial isovaline asymmetry in Murchison and Orgueil occurred during an extended aqueous alteration phase on the meteorite parent bodies. The large asymmetry in isovaline and other alpha-dialkyl amino acids found in altered CI and CM meteorites suggests that amino acids delivered by asteroids, comets, and their fragments would have biased the Earth's prebiotic organic inventory with left-handed molecules before the origin of life. PMID- 19289827 TI - Imprecise probability assessment of tipping points in the climate system. AB - Major restructuring of the Atlantic meridional overturning circulation, the Greenland and West Antarctic ice sheets, the Amazon rainforest and ENSO, are a source of concern for climate policy. We have elicited subjective probability intervals for the occurrence of such major changes under global warming from 43 scientists. Although the expert estimates highlight large uncertainty, they allocate significant probability to some of the events listed above. We deduce conservative lower bounds for the probability of triggering at least 1 of those events of 0.16 for medium (2-4 degrees C), and 0.56 for high global mean temperature change (above 4 degrees C) relative to year 2000 levels. PMID- 19289828 TI - Natural images dominate in binocular rivalry. AB - Ecological approaches to perception have demonstrated that information encoding by the visual system is informed by the natural environment, both in terms of simple image attributes like luminance and contrast, and more complex relationships corresponding to Gestalt principles of perceptual organization. Here, we ask if this optimization biases perception of visual inputs that are perceptually bistable. Using the binocular rivalry paradigm, we designed stimuli that varied in either their spatiotemporal amplitude spectra or their phase spectra. We found that noise stimuli with "natural" amplitude spectra (i.e., amplitude content proportional to 1/f, where f is spatial or temporal frequency) dominate over those with any other systematic spectral slope, along both spatial and temporal dimensions. This could not be explained by perceived contrast measurements, and occurred even though all stimuli had equal energy. Calculating the effective contrast following attenuation by a model contrast sensitivity function suggested that the strong contrast dependency of rivalry provides the mechanism by which binocular vision is optimized for viewing natural images. We also compared rivalry between natural and phase-scrambled images and found a strong preference for natural phase spectra that could not be accounted for by observer biases in a control task. We propose that this phase specificity relates to contour information, and arises either from the activity of V1 complex cells, or from later visual areas, consistent with recent neuroimaging and single-cell work. Our findings demonstrate that human vision integrates information across space, time, and phase to select the input most likely to hold behavioral relevance. PMID- 19289829 TI - A microraptorine (Dinosauria-Dromaeosauridae) from the Late Cretaceous of North America. AB - The fossil assemblages of the Late Cretaceous of North America are dominated by large-bodied dinosaur species. Associated skeletons of small dinosaurs are exceedingly rare, and small (<10 kg) carnivorous theropods have not previously been reported from these beds. Here, we describe a small dromaeosaurid from the 75-million-year-old Dinosaur Park Formation of Alberta, Canada. Hesperonychus elizabethae gen. et sp. nov. is represented by a pelvic girdle from an animal weighing approximately 1,900 g. Despite its size, the pubes and ilia are coossified, indicating that the animal was somatically mature. This is the smallest carnivorous, nonavian dinosaur known from North America. Phylogenetic analysis of Hesperonychus reveals that it is not closely related to previously described North American dromaeosaurids. Instead, Hesperonychus is a member of the dromaeosaurid clade Microraptorinae, a group containing the 4-winged Microraptor and the feathered Sinornithosaurus, both from the Lower Cretaceous Jehol Group of China. Hesperonychus is the youngest known member of this lineage, extending the temporal range of the clade by 45 million years, and it is the first microraptorine known from North America, providing further evidence for an affinity between the dinosaur faunas of North America and Asia. Study of fossil collections from the Dinosaur Park and Oldman formations of Alberta has revealed numerous isolated bones of small, basal dromaeosaurids, which are tentatively referred to Hesperonychus. These fossils suggest that small dromaeosaurids were a significant component of the carnivore community in this Late Cretaceous biota. PMID- 19289830 TI - Linking international agricultural research knowledge with action for sustainable development. AB - We applied an innovation framework to sustainable livestock development research projects in Africa and Asia. The focus of these projects ranged from pastoral systems to poverty and ecosystems services mapping to market access by the poor to fodder and natural resource management to livestock parasite drug resistance. We found that these projects closed gaps between knowledge and action by combining different kinds of knowledge, learning, and boundary spanning approaches; by providing all partners with the same opportunities; and by building the capacity of all partners to innovate and communicate. PMID- 19289831 TI - The two-component system QseEF and the membrane protein QseG link adrenergic and stress sensing to bacterial pathogenesis. AB - Bacterial pathogens sense host cues to activate expression of virulence genes. Most of these signals are sensed through histidine kinases (HKs), which comprise the main sensory mechanism in bacteria. The host stress hormones epinephrine (Epi) and norepinephrine are sensed through the QseC HK, which initiates a complex signaling cascade to regulate virulence gene expression in enterohemorrhagic Escherichia coli (EHEC). Epi signaling through QseC activates expression of the genes encoding the QseEF 2-component system. QseE is an HK, and QseF is a response regulator. Here, we show that QseE is a second bacterial adrenergic receptor that gauges the stress signals Epi, sulfate, and phosphate. The qseEF genes are organized within an unusual operonic structure, in that a gene is encoded between qseE and qseF. This gene was renamed qseG, and it was shown to encode an outer membrane (OM) protein. EHEC uses a type III secretion system (TTSS) to translocate effector proteins to the epithelial cells that rearrange the host cytoskeleton to form pedestal-like structures that cup the bacterium. QseE, QseG, and QseF are necessary for pedestal formation. Although QseE and QseF are involved in the transcriptional control of genes necessary for pedestal formation, QseG is necessary for translocation of effectors into epithelial cells. QseG is an OM protein necessary for translocation of TTSS effectors that also works in conjunction with a 2-component signaling system that senses host stress signals. PMID- 19289832 TI - EZH2 is a mediator of EWS/FLI1 driven tumor growth and metastasis blocking endothelial and neuro-ectodermal differentiation. AB - Ewing tumors (ET) are highly malignant, localized in bone or soft tissue, and are molecularly defined by ews/ets translocations. DNA microarray analysis revealed a relationship of ET to both endothelium and fetal neural crest. We identified expression of histone methyltransferase enhancer of Zeste, Drosophila, Homolog 2 (EZH2) to be increased in ET. Suppressive activity of EZH2 maintains stemness in normal and malignant cells. Here, we found EWS/FLI1 bound to the EZH2 promoter in vivo, and induced EZH2 expression in ET and mesenchymal stem cells. Down regulation of EZH2 by RNA interference in ET suppressed oncogenic transformation by inhibiting clonogenicity in vitro. Similarly, tumor development and metastasis was suppressed in immunodeficient Rag2(-/-)gamma(C)(-/-) mice. EZH2-mediated gene silencing was shown to be dependent on histone deacetylase (HDAC) activity. Subsequent microarray analysis of EZH2 knock down, HDAC-inhibitor treatment and confirmation in independent assays revealed an undifferentiated phenotype maintained by EZH2 in ET. EZH2 regulated stemness genes such as nerve growth factor receptor (NGFR), as well as genes involved in neuroectodermal and endothelial differentiation (EMP1, EPHB2, GFAP, and GAP43). These data suggest that EZH2 might have a central role in ET pathology by shaping the oncogenicity and stem cell phenotype of this tumor. PMID- 19289833 TI - Resistance to CCR5 inhibitors caused by sequence changes in the fusion peptide of HIV-1 gp41. AB - We have investigated the mechanism of resistance of a HIV type 1 (HIV-1) R5 primary isolate, D1/85.16, to the small molecule CCR5 inhibitor, vicriviroc (VVC). Unlike other viruses resistant to this class of compound, D1/85.16 lacks sequence changes in the V3 region of the gp120 surface glycoprotein. Inspection of env sequences from D1/85.16 compared with those derived from the parental, inhibitor-sensitive virus, CC1/85, revealed a cluster of 3 conservative changes in the fusion peptide (FP) of the gp41 transmembrane glycoprotein that tracked with the resistance phenotype. Studies with engineered Env-chimeric and point substituted viruses confirmed that these 3 FP residues were substantially responsible for VVC resistance without altering coreceptor usage, as assessed in both peripheral blood mononuclear cells and the TZM-bl cell line. VVC resistance is manifested differently in the 2 cell types, and there are assay-dependent complexities to the dose-response curves for the engineered resistant viruses. To explain them, we created a model for resistance and generated theoretical VVC inhibition curves that closely mimic the experimental data for the resistant viruses. The basis for the model is the existence of distinct forms of CCR5, with varying affinities for small molecule CCR5 inhibitors that are presumed to be present in different proportions on different cell types, and are used selectively by resistant HIV-1 variants when ligated with an inhibitor. Together, the experimental results and theoretical model may help understand how HIV-1 uses CCR5 to enter target cells under various conditions. PMID- 19289834 TI - Designed amphiphilic peptide forms stable nanoweb, slowly releases encapsulated hydrophobic drug, and accelerates animal hemostasis. AB - How do you design a peptide building block to make 2-dimentional nanowebs and 3 dimensional fibrous mats? This question has not been addressed with peptide self assembling nanomaterials. This article describes a designed 9-residue peptide, N Pro-Ser-Phe-Cys-Phe-Lys-Phe-Glu-Pro-C, which creates a strong fishnet-like nanostructure depending on the peptide concentrations and mechanical disruptions. This peptide is intramolecularly amphiphilic because of a single pair of ionic residues, Lys and Glu, at one end and nonionic residues, Phe, Cys, and Phe, at the other end. Circular dichroism and Fourier transform infrared spectroscopy analysis demonstrated that this peptide adopts stable beta-turn and beta-sheet structures and self-assembles into hierarchically arranged supramolecular aggregates in a concentration-dependent fashion, demonstrated by atomic force microscopy and electron microscopy. At high concentrations, the peptide dominantly self-assembled into globular aggregates that were extensively connected with each other to form "beads-on-a-thread" type nanofibers. These long nanofibers were extensively branched and overlapped to form a self-healing peptide hydrogel consisting of >99% water. This peptide can encapsulate the hydrophobic model drug pyrene and slowly release pyrene from coated microcrystals to liposomes. It can effectively stop animal bleeding within 30 s. We proposed a plausible model to interpret the intramolecular amphiphilic self-assembly process and suggest its importance for the future development of new biomaterials for drug delivery and regenerative medicine. PMID- 19289835 TI - Induction and regulatory function of miR-9 in human monocytes and neutrophils exposed to proinflammatory signals. AB - Inflammation involves a coordinated, sequential, and self limiting sequence of events controlled by positive and negative regulatory mechanisms. Recent studies have shown that microRNAs (miRNAs), an evolutionarily conserved class of endogenous 22-nucleotide noncoding RNAs, contribute to the regulation of inflammation by repressing gene expression at the posttranscriptional level. In this study, we characterize the profile of miRNAs induced by LPS in human polymorphonuclear neutrophils (PMN) and monocytes. In particular, we identify miR 9 as the only miRNA (among 365 analyzed) up-regulated in both cell types after TLR4 activation. miR-9 is also induced by TLR2 and TLR7/8 agonists and by the proinflammatory cytokines TNF-alpha and IL-1beta, but not by IFNgamma. Among the 3 different genes encoding miR-9 precursors in humans, we show that LPS selectively induces the transcription of miR-9-1 located in the CROC4 locus, in a MyD88- and NF-kappaB-dependent manner. In PMN and monocytes, LPS regulates NFKB1 at both the transcriptional and posttranscriptional levels, and a conserved miR-9 seed sustained a miR-9-dependent inhibition of the NFKB1 transcript. Overall, these data suggest that TLR4-activated NF-kappaB rapidly increases the expression of miR-9 that operates a feedback control of the NF-kappaB-dependent responses by fine tuning the expression of a key member of the NF-kappaB family. PMID- 19289836 TI - Amplification of the North American "Dust Bowl" drought through human-induced land degradation. AB - The "Dust Bowl" drought of the 1930s was highly unusual for North America, deviating from the typical pattern forced by "La Nina" with the maximum drying in the central and northern Plains, warm temperature anomalies across almost the entire continent, and widespread dust storms. General circulation models (GCMs), forced by sea surface temperatures (SSTs) from the 1930s, produce a drought, but one that is centered in southwestern North America and without the warming centered in the middle of the continent. Here, we show that the inclusion of forcing from human land degradation during the period, in addition to the anomalous SSTs, is necessary to reproduce the anomalous features of the Dust Bowl drought. The degradation over the Great Plains is represented in the GCM as a reduction in vegetation cover and the addition of a soil dust aerosol source, both consequences of crop failure. As a result of land surface feedbacks, the simulation of the drought is much improved when the new dust aerosol and vegetation boundary conditions are included. Vegetation reductions explain the high temperature anomaly over the northern U.S., and the dust aerosols intensify the drought and move it northward of the purely ocean-forced drought pattern. When both factors are included in the model simulations, the precipitation and temperature anomalies are of similar magnitude and in a similar location compared with the observations. Human-induced land degradation is likely to have not only contributed to the dust storms of the 1930s but also amplified the drought, and these together turned a modest SST-forced drought into one of the worst environmental disasters the U.S. has experienced. PMID- 19289837 TI - The DNA-damage effector checkpoint kinase 1 is essential for chromosome segregation and cytokinesis. AB - Defective genome maintenance mechanisms, involving DNA repair and cell-cycle checkpoint pathways, initiate genetic instability in many sporadic and hereditary cancers. The DNA damage effector Checkpoint kinase 1 (Chk1) is a critical component of DNA replication, intra-S phase, and G(2)/M phase checkpoints and a recently reported mitotic spindle-assembly checkpoint. Here, we report for the first time that haploinsufficiency of Chk1 in mice resulted in multiple mitotic defects and enhanced binucleation. We observed that Aurora B, a critical cytokinetic regulator and a recently identified Chk1 substrate, was mislocalized in mitotic Chk1(+/-) mammary epithelia. Chk1 also exhibited distinct mitotic localization patterns and was active during unperturbed mitosis and cytokinesis in mammalian cells. Active Chk1 expression was not dependent on treatment with spindle poisons such as colcemid during mitosis and cytokinesis. Furthermore, two different complementary approaches demonstrated that abrogation of Chk1 in mitotic mammalian cells resulted in cytokinetic regression and binucleation, increased chromosome lagging and/or nondisjunction, and abnormal localization of Aurora B at late mitotic structures. Thus, Chk1 is a multifunctional kinase that serves as a nexus between the DNA damage response and the mitotic exit pathways during cell-cycle progression to prevent genomic instability and cancer. PMID- 19289838 TI - Molecular recognition and self-assembly special feature: Integrative self-sorting is a programming language for high level self-assembly. AB - Starting from the basis of a simple 4-component self-sorting system of crown ethers and ammonium ions, we design 6 building blocks in which 2 identical or different binding sites are incorporated. These building blocks can be mixed in many different ways to yield quite distinctly different pseudorotaxane assemblies. The self-sorting process integrates all building blocks in specific places so that this approach permits us to exert positional control and can widely influence the resulting assemblies with respect to the details of their structures. At maximum, we report quadruply interlocked species with up to 5 subunits that form specific assemblies. Although NMR methods are limited to the analysis of simpler complexes, ESI-MS and, in particular, tandem mass spectrometry is highly useful to analyze the assemblies' connectivities. PMID- 19289839 TI - Identification of targets of the Wnt pathway destruction complex in addition to beta-catenin. AB - The proteasomal degradation of beta-catenin mediated by the glycogen synthase kinase 3beta (GSK3beta) and destruction complex is the central step in the canonical Wnt signaling pathway. However, that there are branches of Wnt signaling pathways that do not depend on beta-catenin/Tcf-mediated transcription activation has long been understood. In this study, we hypothesized that there are many more GSK3 and destruction complex-dependent proteolytic target proteins that mediate Wnt signaling in the cell. To test this hypothesis, we have developed and carried out a screen for such candidate proteins using an in vitro expression cloning technique and biochemical reconstitution of Wnt signaling in Xenopus egg cytoplasmic extracts. Forty-two proteins have been identified as potential candidates for GSK3-regulated phosphorylation, proteasomal degradation, or both, of which 12 are strong candidates for Wnt-pathway-regulated degradation. Some of them have been reported to interact with beta-catenin and implicated in the canonical Wnt signaling pathway, and other targets identified include proteins with various cellular functions such as RNA processing, cytoskeletal dynamics, and cell metabolism. Thus, we propose that Wnt/GSK3/destruction complex signaling regulates multiple target proteins to control a broad range of cellular activities in addition to beta-catenin-mediated transcription activation. PMID- 19289840 TI - MetJ repressor interactions with DNA probed by in-cell NMR. AB - Atomic level characterization of proteins and other macromolecules in the living cell is challenging. Recent advances in NMR instrumentation and methods, however, have enabled in-cell studies with prospects for multidimensional spectral characterization of individual macromolecular components. We present NMR data on the in-cell behavior of the MetJ repressor from Escherichia coli, a protein that regulates the expression of genes involved in methionine biosynthesis. NMR studies of whole cells along with corresponding studies in cell lysates and in vitro preparations of the pure protein give clear evidence for extensive nonspecific interactions with genomic DNA. These interactions can provide an efficient mechanism for searching out target sequences by reducing the dependence on 3-dimensional diffusion through the crowded cellular environment. DNA provides the track for MetJ to negotiate the obstacles inherent in cells and facilitates locating and binding specific repression sites, allowing for timely control of methionine biosynthesis. PMID- 19289841 TI - Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound. AB - Canonical transient receptor potential (TRPC) channels control influxes of Ca(2+) and other cations that induce diverse cellular processes upon stimulation of plasma membrane receptors coupled to phospholipase C (PLC). Invention of subtype specific inhibitors for TRPCs is crucial for distinction of respective TRPC channels that play particular physiological roles in native systems. Here, we identify a pyrazole compound (Pyr3), which selectively inhibits TRPC3 channels. Structure-function relationship studies of pyrazole compounds showed that the trichloroacrylic amide group is important for the TRPC3 selectivity of Pyr3. Electrophysiological and photoaffinity labeling experiments reveal a direct action of Pyr3 on the TRPC3 protein. In DT40 B lymphocytes, Pyr3 potently eliminated the Ca(2+) influx-dependent PLC translocation to the plasma membrane and late oscillatory phase of B cell receptor-induced Ca(2+) response. Moreover, Pyr3 attenuated activation of nuclear factor of activated T cells, a Ca(2+) dependent transcription factor, and hypertrophic growth in rat neonatal cardiomyocytes, and in vivo pressure overload-induced cardiac hypertrophy in mice. These findings on important roles of native TRPC3 channels are strikingly consistent with previous genetic studies. Thus, the TRPC3-selective inhibitor Pyr3 is a powerful tool to study in vivo function of TRPC3, suggesting a pharmaceutical potential of Pyr3 in treatments of TRPC3-related diseases such as cardiac hypertrophy. PMID- 19289842 TI - Antibodies specifically targeting a locally misfolded region of tumor associated EGFR. AB - Epidermal Growth Factor Receptor (EGFR) is involved in stimulating the growth of many human tumors, but the success of therapeutic agents has been limited in part by interference from the EGFR on normal tissues. Previously, we reported an antibody (mab806) against a truncated form of EGFR found commonly in gliomas. Remarkably, it also recognizes full-length EGFR on tumor cells but not on normal cells. However, the mechanism for this activity was unclear. Crystallographic structures for Fab:EGFR(287-302) complexes of mAb806 (and a second, related antibody, mAb175) show that this peptide epitope adopts conformations similar to those found in the wtEGFR. However, in both conformations observed for wtEGFR, tethered and untethered, antibody binding would be prohibited by significant steric clashes with the CR1 domain. Thus, these antibodies must recognize a cryptic epitope in EGFR. Structurally, it appeared that breaking the disulfide bond preceding the epitope might allow the CR1 domain to open up sufficiently for antibody binding. The EGFR(C271A/C283A) mutant not only binds mAb806, but binds with 1:1 stoichiometry, which is significantly greater than wtEGFR binding. Although mAb806 and mAb175 decrease tumor growth in xenografts displaying mutant, overexpressed, or autocrine stimulated EGFR, neither antibody inhibits the in vitro growth of cells expressing wtEGFR. In contrast, mAb806 completely inhibits the ligand-associated stimulation of cells expressing EGFR(C271A/C283A). Clearly, the binding of mAb806 and mAb175 to the wtEGFR requires the epitope to be exposed either during receptor activation, mutation, or overexpression. This mechanism suggests the possibility of generating antibodies to target other wild-type receptors on tumor cells. PMID- 19289843 TI - Molecular recognition and self-assembly special feature: Supramolecular polymer formed by reversible self-assembly of tetrakisporphyrin. AB - S-shaped tetrakisporphyrin 2 forms supramolecular polymeric assemblies via a complementary affinity of its bisporphyrin units in solution. The self association constant determined by applying the isodesmic model is >10(6) L mol( 1), which suggests that a sizable polymer forms at millimolar concentrations at room temperature. The electron deficient aromatic guest (TNF) binds within the molecular clefts provided by the bisporphyrin units via a charge-transfer interaction. This guest complexation completely disrupts supramolecular polymeric assembly. The long, fibrous fragments of the polymeric assemblies were characterized by atomic-force microscopy imaging of a film cast on a mica surface. The polymeric assemblies have lengths of >1mum and show a coiled structure with a higher level of organization. The approach discussed in this report concerning the quick preparation of supramolecular polymeric assemblies driven by noncovalent forces sets the stage for the preparation of a previously undescribed class of macromolecular porphyrin architectures. PMID- 19289844 TI - Impaired recollection but spared familiarity in patients with extended hippocampal system damage revealed by 3 convergent methods. AB - To understand recognition memory, the detection of stimulus repetition, it first is necessary to resolve the debate between 2 fundamentally different models of recognition. Contemporary single-process models assume that recognition memory relies solely on the neural system required for the recall of prior events. Dual process models assume that recognition comprises 2 independent forms of memory: one supports recall, and the other detects repeated stimuli by signaling their familiarity, the feeling of previous occurrence without the recall of any associated information. These 2 models were contrasted in patients who had undergone surgical removal of a colloid cyst, a condition associated with memory loss when accompanied by fornix and/or mammillary body atrophy. Comparisons were made between 2 groups of 9 patients that differed only with respect to the extent of mammillary body atrophy. Only the more atrophied group was impaired on tests of recall, but both groups showed normal recognition levels on a task that equates recall and recognition performance in normal participants. To explore the nature of this spared recognition, we estimated recall-based recognition and familiarity-based recognition using 3 distinct methods: self-report, receiver operating characteristics, and structural equation modeling. All 3 methods showed impaired recall-based recognition accompanied by intact familiarity in the most atrophied group, as predicted only by dual-process models. When structural equation modeling was applied to all 62 colloid cyst patients, the recall/familiarity dual-process model best explained the patients' memory pattern. The convergent evidence that mammillary body atrophy impairs recall but spares familiarity-based recognition appears irreconcilable with single-process models. PMID- 19289845 TI - MicroRNA expression profiles predictive of human renal allograft status. AB - Immune rejection of organ transplants is a life-threatening complication and is exemplified by alterations in the expression of protein-encoding genes. Because microRNAs (miRNAs) regulate the expression of genes implicated in adaptive immunity, we investigated whether acute rejection (AR) is associated with alterations in miRNA expression within allografts and whether expression profiles are diagnostic of AR and predict allograft function. Seven of 33 renal allograft biopsies (12 AR and 21 normal) were profiled using microfluidic cards containing 365 mature human miRNAs (training set), and a subset of differentially expressed miRNAs were quantified in the remaining 26 allograft biopsies (validation set). We found a strong association between intragraft expression of miRNAs and messenger RNAs (mRNAs), and that AR, and renal allograft function, could be predicted with a high level of precision using intragraft levels of miRNAs. Our investigation of miRNA expression in normal human peripheral blood mononuclear cells (PBMCs) showed that miRNAs (miR-142-5p, -155, and -223) overexpressed in AR biopsies are highly expressed in PBMCs, and that stimulation with the mitogen phytohaemagglutinin results in an increase in the abundance of miR-155 and a decrease in miR-223 and let-7c. Quantification of miRNAs in primary cultures of human renal epithelial cells (HRECs) showed that miR-30a-3p, -10b, and let-7c are highly expressed in HRECs, and that stimulation results in a decreased expression of miR-30a-3p. Our studies, in addition to suggesting a cellular basis for the altered intragraft expression of miRNAs, propose that miRNA expression patterns may serve as biomarkers of human renal allograft status. PMID- 19289847 TI - Manipulability of comparative tests. AB - Multiple self-proclaimed experts claim that they know the probabilities of future events. A tester does not know the odds of future events and she also does not know whether, among the multiple experts, there are some who do know the relevant probabilities. So the tester requires each expert to announce, before any data are observed, the probabilities of all future events. A test either rejects or does not reject each expert based on the observed data and the profile of the probabilities announced by the experts. We assume that the test controls for the type I error of rejecting the true probabilities. However, consider the case in which all experts are uninformed (i.e., they do not know anything about true probabilities). We show that they can still independently produce false forecasts that are likely to both pass the test, no matter how the data evolve in the future. Hence, the data may not suffice to effectively discredit uninformed, but strategic, experts. PMID- 19289846 TI - MUC1-induced alterations in a lipid metabolic gene network predict response of human breast cancers to tamoxifen treatment. AB - The mucin 1 (MUC1) oncoprotein is aberrantly overexpressed in human breast cancers. Although MUC1 modulates the activity of estrogen receptor alpha (ER), there is no information regarding the effects of MUC1 on global gene expression patterns and the potential role of MUC1-induced genes in predicting outcome for breast cancer patients. We have developed an experimental model of MUC1-induced transformation that has identified the activation of genes involved in cholesterol and fatty acid metabolism. A 38-gene set of experimentally derived MUC1-induced genes associated with lipid metabolism was applied to the analysis of ER(+) breast cancer patients treated with tamoxifen. The results obtained from 2 independent databases demonstrate that patients overexpressing MUC1 and the lipid metabolic pathways are at significantly higher risk for death and recurrence/distant metastasis. By contrast, these genes were not predictive in untreated patients. Furthermore, a positive correlation was found between expression of the 38-gene set and the ER signaling pathway. These findings indicate that (i) MUC1 regulates cholesterol and fatty acid metabolism, and (ii) activation of these pathways in ER(+) breast cancers predicts failure to tamoxifen treatment. PMID- 19289848 TI - Reconstructing web evolution and spider diversification in the molecular era. AB - The evolutionary diversification of spiders is attributed to spectacular innovations in silk. Spiders are unique in synthesizing many different kinds of silk, and using silk for a variety of ecological functions throughout their lives, particularly to make prey-catching webs. Here, we construct a broad higher level phylogeny of spiders combining molecular data with traditional morphological and behavioral characters. We use this phylogeny to test the hypothesis that the spider orb web evolved only once. We then examine spider diversification in relation to different web architectures and silk use. We find strong support for a single origin of orb webs, implying a major shift in the spinning of capture silk and repeated loss or transformation of orb webs. We show that abandonment of costly cribellate capture silk correlates with the 2 major diversification events in spiders (1). Replacement of cribellate silk by aqueous silk glue may explain the greater diversity of modern orb-weaving spiders (Araneoidea) compared with cribellate orb-weaving spiders (Deinopoidea) (2). Within the "RTA clade," which is the sister group to orb-weaving spiders and contains half of all spider diversity, >90% of species richness is associated with repeated loss of cribellate silk and abandonment of prey capture webs. Accompanying cribellum loss in both groups is a release from substrate constrained webs, whether by aerially suspended webs, or by abandoning webs altogether. These behavioral shifts in silk and web production by spiders thus likely played a key role in the dramatic evolutionary success and ecological dominance of spiders as predators of insects. PMID- 19289849 TI - Cyclophilin 40 is required for microRNA activity in Arabidopsis. AB - Loss-of-function mutations of SQUINT (SQN)-which encodes the Arabidopsis orthologue of cyclophilin 40 (CyP40)-cause the precocious expression of adult vegetative traits, an increase in carpel number, and produce abnormal spacing of flowers in the inflorescence. Here we show that the vegetative phenotype of sqn is attributable to the elevated expression of miR156-regulated members of the SPL family of transcription factors and provide evidence that this defect is a consequence of a reduction in the activity of ARGONAUTE1 (AGO1). Support for this latter conclusion was provided by the phenotypic similarity between hypomorphic alleles of AGO1 and null alleles of SQN and by the genetic interaction between sqn and these alleles. Our results suggest that AGO1, or an AGO1-interacting protein, is a major client of CyP40 and that miR156 and its targets play a central role in the regulation of vegetative phase change in Arabidopsis. PMID- 19289850 TI - Phase TEM for biological imaging utilizing a Boersch electrostatic phase plate: theory and practice. AB - A Boersch electrostatic phase plate (BEPP) used in a transmission electron microscope (TEM) system can provide tuneable phase shifts and overcome the low contrast problem for biological imaging. Theoretically, a pure phase image with a high phase contrast can be obtained using a BEPP. However, a currently available TEM system utilizing a BEPP cannot achieve sufficiently high phase efficiency for biological imaging, owing to the practical conditions. The low phase efficiency is a result of the blocking of partial unscattered electrons by BEPP, and the contribution of absorption contrast. The fraction of blocked unscattered beam is related to BEPP dimensions and to divergence of the illumination system of the TEM. These practical issues are discussed in this paper. Phase images of biological samples (negatively stained ferritin) obtained by utilizing a BEPP are reported, and the phase contrast was found to be enhanced by a factor of approximately 1.5, based on the calculation using the Rose contrast criterion. The low gain in phase contrast is consistent with the expectation from the current TEM/BEPP system. A new generation of phase TEM utilizing BEPP and designed for biological imaging with a high phase efficiency is proposed. PMID- 19289851 TI - Smart specimen preparation for freeze substitution and serial ultrathin sectioning of yeast cells. AB - A smart and efficient method for freeze substitution and serial sectioning of yeast cells is described. Yeast cells were placed in a single layer between two copper disks, rapidly frozen, freeze substituted and embedded in an epoxy resin. The cell layer was re-embedded by the same resin, the surface trimmed leaving 1 mum above the cell layer, and serially sectioned. The sections were collected on the two-slit grids and placed on a Formvar film mounted to cover the holes of an aluminum supporting rack. The grids were removed from the rack, stained together using a silicon tube and observed in a transmission electron microscope. The images of yeast cells observed were clear and natural, and would be useful for a detailed 3D structural analysis such as structome. PMID- 19289852 TI - Imatinib mesylate as salvage therapy for refractory sclerotic chronic graft versus-host disease. AB - Imatinib is a promising candidate for the treatment of fibrotic diseases. This retrospective study evaluated the use of imatinib for the treatment of refractory sclerotic chronic graft-versus-host disease in 14 patients with different hematologic malignancies. Imatinib was started at a median of 44 months after transplantation (range, 16-119 months after transplantation) and was administered for a median of 5.9 months from time of initiation (range, 2.1-74 months from time of initiation). With a median overall follow-up of 11.6 months from time of initiation (range, 4.1-74 months from time of initiation) of imatinib, 4 patients (29%) had to stop imatinib because of drug intolerance. All other adverse reactions were of mild-to-moderate grade and could be managed symptomatically. Overall, 7 patients responded to imatinib (50%; 95% confidence interval, 24%-76%) with 4 patients improving their Rodman score more than or equal to 90%. In addition, imatinib therapy allowed for a significant reduction of corticosteroid dosage. Despite its limited size, this cohort suggests some beneficial activity of imatinib in sclerotic chronic graft-versus-host disease, warranting further prospective investigations. PMID- 19289853 TI - Endogenous IL-17 contributes to reduced tumor growth and metastasis. AB - It has been reported that ectopically expressed interleukin-17 (IL-17) in tumor cells suppresses tumor progression through enhanced antitumor immunity in immune competent mice or promote tumor progression through an increase in inflammatory angiogenesis in immune-deficient mice. The role of endogenous IL-17 in tumor immunity remains undefined. Here we showed that tumor growth and lung metastasis were enhanced in IL-17-deficient mice, associated with decreased interferon gamma(+) natural killer cells and tumor specific interferon-gamma(+) T cells in the tumor draining lymph nodes and tumors. Together with the published data showing that in vitro transforming growth factor-beta and IL-6-polarized Th17 cells induce tumor regression, our work supports the notion that endogenous IL-17 or/and Th17 cells may play a protective role in tumor immunity. PMID- 19289855 TI - The favorable impact of CEBPA mutations in patients with acute myeloid leukemia is only observed in the absence of associated cytogenetic abnormalities and FLT3 internal duplication. AB - Mutations of the CCAAT/enhancer binding protein alpha (CEBPA) gene have been associated with a favorable outcome in patients with acute myeloid leukemia (AML), but mainly in those with a normal karyotype. Here, we analyzed the impact of associated cytogenetic abnormalities or bad-prognosis fms-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD) in 53 patients with CEBPA(+) de novo AML treated in the Acute Leukemia French Association trials. We found that only those with a normal karyotype and no FLT3-ITD displayed the expected favorable outcome. In this context, relapse-free, disease-free, and overall survival were significantly longer than in corresponding patients without the CEBPA mutation (P = .035, .016, and .047, respectively). This was not observed in the context of an abnormal karyotype or associated FLT3-ITD. Furthermore, after adjustment on age, trial, and mutation type, these features were independently predictive of shorter overall survival in the subset of patients with CEBPA(+) AML (multivariate hazard ratio = 2.7; 95% confidence interval, 1.08-6.7; and 2.9; 95% confidence interval, 1.01-8.2; with P = .034 and .05, for abnormal karyotype and FLT3-ITD, respectively). PMID- 19289854 TI - Therapeutic targeting of MLL. AB - Treatment of hematologic malignancies is evolving from a uniform approach to targeted therapies directed at the underlying molecular abnormalities of disease. The mixed lineage leukemia (MLL) proto-oncogene is a recurrent site of genetic rearrangements in acute leukemias; and since its discovery in 1992, many advances have been made in understanding its role in leukemogenesis. A variety of MLL translocation partners have been described, and detailed structure/function studies have identified functional domains that are required for transformation. Proteins associated with the MLL core complex or its fusion partners have been isolated and characterized for their critical roles in leukemia pathogenesis. Downstream mediators of MLL transcriptional regulation and multiple collaborating signaling pathways have been described and characterized. These advances in our understanding of MLL-related leukemogenesis provide a foundation for ongoing and future efforts to develop novel therapeutic strategies that will hopefully result in better treatment outcomes. PMID- 19289856 TI - Exclusive expression of proteasome subunit {beta}5t in the human thymic cortex. AB - The ubiquitin-proteasome pathway, which degrades intracellular proteins, is involved in numerous cellular processes, including the supply of immunocompetent peptides to the antigen presenting machinery. Proteolysis by proteasomes is conducted by three beta subunits, beta1, beta2, and beta5, of the 20S proteasome. Recently, a novel beta subunit expressed exclusively in cortical thymic epithelial cells was discovered in mice. This subunit, designated beta5t, is a component of the thymoproteasome, a specialized type of proteasomes implicated in thymic positive selection. In this study, we show that, like its mouse counterpart, human beta5t is expressed exclusively in the thymic cortex. Human beta5t was expressed in approximately 80% of cortical thymic epithelial cells and some cortical dendritic cells. Human beta5t was incorporated into proteasomes with two other catalytically active beta subunits beta1i and beta2i, forming 20S proteasomes with subunit compositions characteristic of thymoproteasomes. The present study demonstrates, for the first time, the existence of thymoproteasomes in the human thymic cortex, indicating that thymoproteasome function is likely conserved between humans and mice. PMID- 19289858 TI - Association of endogenous sex hormones with diabetes and impaired fasting glucose in men: multi-ethnic study of atherosclerosis. AB - OBJECTIVE: To assess associations of sex hormones with impaired fasting glucose (IFG) and type 2 diabetes in men. RESEARCH DESIGN AND METHODS: A total of 3,156 African American, Non-Hispanic white, Hispanic, and Chinese-American men aged 45 84 years who participated in the baseline visit of the Multi-Ethnic Study of Atherosclerosis (MESA) were included. Odd sratios and 95% CIs for type 2 diabetes and IFG compared with normal fasting glucose for quartiles of hormones were estimated. RESULTS: After adjusting for age, ethnicity, BMI, and waist circumference, IFG and diabetes were associated inversely with total testosterone and sex hormone-binding globulin (SHBG) and positively with estradiol (E2). Dehydroepiandrosterone was positively associated with IFG but not with diabetes. Associations did not differ across ethnic groups. CONCLUSIONS: Regardless of obesity, total testosterone and SHBG were associated inversely and E2 was associated positively with IFG and diabetes in men. Further research is warranted to better understand the underlying biological mechanisms. PMID- 19289857 TI - Efficacy and safety of the human glucagon-like peptide-1 analog liraglutide in combination with metformin and thiazolidinedione in patients with type 2 diabetes (LEAD-4 Met+TZD). AB - OBJECTIVE: To determine the efficacy and safety of liraglutide (a glucagon-like peptide-1 receptor agonist) when added to metformin and rosiglitazone in type 2 diabetes. RESEARCH DESIGN AND METHODS: This 26-week, double-blind, placebo controlled, parallel-group trial randomized 533 subjects (1:1:1) to once-daily liraglutide (1.2 or 1.8 mg) or liraglutide placebo in combination with metformin (1 g twice daily) and rosiglitazone (4 mg twice daily). Subjects had type 2 diabetes, A1C 7-11% (previous oral antidiabetes drug [OAD] monotherapy >or=3 months) or 7-10% (previous OAD combination therapy >or=3 months), and BMI 0.05, respectively, Mann-Whitney's U-test). In this program, participation in competitive employment may not be associated with the severity of psychotic symptoms, disability of daily life or subjective QOL but may be promoted by the disclosure of disabilities, shorter initial working-time and higher education. PMID- 19289877 TI - Exposure to traumatic events among adolescents in four nations. AB - BACKGROUND: Although studies indicate that adolescents, like adults, might develop posttraumatic stress disorder after exposure to traumatic events, the research on this age group is still sparse. METHOD: In three national representative samples and one national total sample of 1,206 8th and 9th-grade students with a mean age of 14.5 years, the prevalence of 19 potential traumatizing and distressing events were reported, along with the psychological impact of these events. FINDINGS: Ninety percent of the adolescents had been exposed to at least one event. The most common events were the death of a family member, threat of violence, bullying, near-drowning, and traffic accidents. Gender was associated with specific events. The estimated lifetime prevalence of posttraumatic stress disorder (PTSD) in the total sample was 14.6 percent, whereas another 13 percent reached a subclinical level of PTSD. Following exposure, females suffered from PTSD two and a half times more often than males. The relative risk for PTSD given a specific event is described. Being exposed to multiple traumatic events was associated with an increase in PTSD. Cultural differences were found in prevalence of PTSD, exposure to specific events and in the female male ratio in PTSD. CONCLUSION: The findings indicate substantial mental health problems in adolescents that are associated with various types of victimization. PMID- 19289878 TI - Prevalence of traumatic events and posttraumatic stress symptoms in a student sample in Poland. AB - The study investigated the prevalence of traumatic events and posttraumatic symptoms among university level students in Poland. Data was collected from 475 students: 69% women and 30% men, mean age 22.9. The measures included SLESQ, Mississippi-C Scale, IES and BDI. At least one traumatic event (according to DSM IV) was reported by 75.6% of the studied group. Prevalence of traumatic events was higher for men than for women. Life threatening accidents, child physical abuse, traumatic bereavement, witnessing death/assault and adult physical assault/abuse were the most commonly experienced events in the whole group. There were differences in prevalence rates of specific types of traumatic events between men and women. The level of posttraumatic events between groups with different levels of exposure to trauma was analysed, as well as between the group of persons who experienced particular types of traumatic events as compared with the group of subjects with no exposure to this type of trauma. PMID- 19289879 TI - Comparing psychological responses of internally displaced and non-displaced Turkish Cypriots. AB - During the 1963-1964 ethnic conflict and 1974 war in Cyprus, many Turkish Cypriots were displaced by Greek Cypriot forces. The psychological condition of Turkish Cypriots after these conflicts has not been studied to the present day. At the time of the Annan Plan Referendum on April 24th 2004, when people on both sides were to decide whether to reunite or not, and when old traumatic events where being discussed in vivid detail, the psychological responses of the internally displaced and non-displaced Turkish Cypriots were investigated. The sample of the study derived from a sample of a larger household survey study conducted on 408 adult people taken randomly from three different districts. People who settled down in Cyprus after 1974 or who had never experienced a war in Cyprus were not included in the study. 129 Turkish Cypriots who experienced either 1963-64 conflict or the 1974 war were included in the present study. 86 of these had been displaced. The first part of the questionnaire that was administered to the subjects included demographic characteristics, war-related traumatic experiences, the level of seriousness, and traumatic incidents resulting from other circumstances. In the second part of the questionnaire, the Traumatic Stress Symptom Checklist (TSSC) and Brief Symptom Inventory (BSI) were used to investigate the symptoms of the post-traumatic process. The outcomes indicates that the internally displaced persons (IDPs) where subjected to traumatic incidents at a higher degree due to killing, displacement, captivity, or killing of family members and relatives. The rate of posttraumatic stress disorder (PTSD) of IDPs is 20%, and is significantly higher than for non displaced persons. The comparison of BSI subscales show that IDPs had a higher level of depression scores than the non-displaced persons. The somatization subscale scores are higher in non-displaced persons. The study reveals a higher frequency of war-related traumatic events in IDPs than in non-displaced people, greater suffering from posttraumatic stress and more negative beliefs about future reunion. PMID- 19289880 TI - Prevalence of PTSD and related factors in communities living in conflictual area: Diyarbakir case. AB - OBJECTIVE: In this study, we aimed to investigate the distribution of Post Traumatic Stress Disorder (PTSD) among adults who were living in the Diyarbakir city center. METHOD: Data was obtained from 708 participants that represented the demographic structure of Diyarbakir. Houses to be visited were determined in collaboration with the Turkish Institute of Statistics. RESULTS: The prevalence of traumatic life experience was 47.9%. Most prevalent traumatic life experiences were forced emigration and witnessing of a case of murder or injury. The lifelong and current PTSD prevalence was 34.9% and 15.1% respectively. We concluded that the prevalence of traumatic experiences and subsequent PTSD was high among people who were living in areas of conflict, and treatment opportunities were inadequate. CONCLUSION: An important finding of this study is the association between the range of prevalence rates of traumatic experiences and risk factors for PTSD in an armed conflict region in Turkey. There is a need for studies that will also include people living in rural areas in order to understand the full picture of problems encountered by those in areas of conflict. Moreover, we believe in the importance of an effective approach of institutional and occupational organizations not to leave these people alone with their traumatic experiences. PMID- 19289881 TI - Justice heals: the impact of impunity and the fight against it on the recovery of severe human rights violations' survivors. AB - Case studies show that traumatized refugees, who are survivors of serious human rights violations, suffer from persisting impunity in their home countries. Ongoing impunity--the inability to overcome the legal protection of the perpetrators assured by impunity laws, incomplete truthfinding, missing integral reparation and a lack of the necessary acknowledgement by society--represents an important obstacle for the recovery of survivors of serious human rights violations. There are reports describing that a high percentage of survivors shows an elevated mental vulnerability caused by impunity. Mental health problems resulting from traumatic experiences can persist or be reactivated by certain events. In particular, family members of the forcibly disappeared suffer from an incomplete mourning due to the uncertain fate of their beloved ones. The ongoing search for the forcibly disappeared under an atmosphere of impunity puts family members under high risk of retraumatization. Studies from other continents also prove that impunity severely affects mental health. Due to the global character of impunity there can be only little evidence about a positive impact of justice on mental health. Nevertheless, a few examples, in particular from Latin America, show that the combined implementation of memory, truth and justice can have a healing impact on those who suffer from trauma. They demonstrate that the fight against impunity is not only a legitimate moral struggle for human rights, but also a basic need for the sustainable recovery of survivors. PMID- 19289882 TI - The role of jurisdiction on persistence of torture in Turkey and public reflections. AB - Torture still is a serious problem in Turkey. There has been a very effective struggle against torture, particularly for effective documentation by health professionals. The Istanbul Protocol has been taken into consideration by the ministry of health, and procedural safeguards with standardized medicolegal documentation had been a part of daily medicolegal practice. However, measures taken on the basis of effective documentation is not sufficient without effective investigation of which the role of jurisdiction is most prominent. Impunity is highly responsible for the persistence of torture, although procedural safeguards on medical examination and medicolegal documentation have had an influence for the decrease of the total number of cases. The Anatolia Agency had distributed information on the total number of punishments in 2007, which drew a more hopeful picture with 5,082 punishments among 33,000 law enforcement officials who had been taken to court. Nevertheless, a press conference held by the Human Rights Foundation of Turkey revealed that this information was not true. They revealed that the cases taken to the court were mostly because of ill treatment instead of torture, and a great majority of these officers had been acquitted between the years 1989-2005. Administrative measures had also been highly insufficient, and among 922 personnel who had been under investigation, only 8 of them had had punishment. The Human Rights Association has had a research on impunity, and only 15% of law enforcement officials who had been taken to the court were ever convicted of their crimes, and all of these punishments had been suspended. Research on cognitive behaviour of judges and prosecutors revealed that they think human rights might threaten the security of the state. This result only clarifies the cause of impunity, thus persistence of torture. The Istanbul University Istanbul Faculty of Medicine, Department of Forensic Medicine, has an outpatient clinic in which torture survivors are examined, and alternative medicolegal documentation is carried out. These patients who were able to have a medicolegal document are observed to benefit from psychotherapy, thus impunity should not only be surmounted for the eradication of torture, but also the healing of the wounds of torture survivors. PMID- 19289883 TI - Chronic pain and PTSD: the Perpetual Avoidance Model and its treatment implications. AB - Posttraumatic Stress Disorder (PTSD) and chronic pain are frequently seen in the aftermath of a traumatic experience. Torture survivors have an increased risk to suffer from these two disorders. Although many studies report high comorbidity,there is still insufficient knowledge on the mechanisms of the development and maintenance of PTSD and chronic pain. After providing an overview of the current literature concerning the comorbidity of these two disorders, we will present the "Perpetual Avoidance Model" (PAM). This model provides an explanation of the reciprocal maintenance of both disorders and offers treatment implications. PMID- 19289884 TI - Asylum seekers in Denmark--a study of health status and grade of traumatization of newly arrived asylum seekers. AB - BACKGROUND: An unknown number of asylum seekers arriving in Denmark have been exposed to torture or have experienced other traumatising events in their country of origin. The health of traumatised asylum seekers, both physically and mentally, is affected upon arrival to Denmark, and time in asylum centres leads to further deterioration in health. METHODS: One hundred forty-two (N=142) newly arrived asylum seekers were examined at Center Sandholm by Amnesty International Danish Medical Group from the 1st of September until the 31st of December 2007. FINDINGS: The asylum seekers came from 33 different countries, primarily representing Afghanistan, Iraq, Iran, Syria, and Chechnya. Of the asylum seekers, 45 percent had been exposed to torture--approximately one-third within the year of arrival to Denmark. Unsystematic blows, personal threats or threats to family, degrading treatment, isolation, and witnessing torture of others were the main torture methods reported. The majority of the asylum seekers had witnessed armed conflict, persecution, and imprisonment. The study showed that physical symptoms were approximately twice as frequent and psychological symptoms were approximately two to three times as frequent among torture survivors as among non tortured asylum seekers. However, even the health of non-tortured asylum seekers was affected. Among the torture survivors, 63 percent fulfilled the criteria for post-traumatic stress disorder, and 30-40 percent of the torture survivors were depressed, in anguish, anxious, and tearful in comparison to 5-10 percent of the non-tortured asylum seekers. Further, 42 percent of torture survivors had torture related scars. INTERPRETATION: Torture survivors amid newly arrived asylum seekers are an extremely vulnerable group, hence examination and inquiry about the torture history is extremely important in order to identify this population to initiate the necessary medical treatment and social assistance. Amnesty International Danish Medical group is currently planning a follow-up study of the present population which will focus on changes in health status during their time in Denmark. PMID- 19289885 TI - A follow up study of allegations of ill-treatment/torture in incommunicado detainees in Spain. Failure of international preventive mechanisms. AB - BACKGROUND: Proper documentation is an important factor in torture prevention, thus making systematic research studies necessary. According to international reports, torture/ill-treatment continues to exist in Spain in relation to Basque people arrested under anti-terrorist legislation (incommunicado detention). To improve the safeguards of these detainees, the European Committee for the Prevention of Torture (CPT) has visited Spain and published recommendations. However, the Spanish Government has not implemented these recommendations. The primary aims of this study were to analyze the methods of torture claimed by Basque incommunicado detainees during 2000-2005 and to compare them with the findings of a previous study (1992-1993), as well as to evaluate the impact of the CPT recommendations. The influence of variables related to police ill treatment were also studied. METHODS: This retrospective study is based on the testimonies given voluntarily by 112 Basques held incommunicado during 2000-2005. Testimonies were collected by a non-governmental organisation. FINDINGS: Threats (91 percent) and beatings (89 percent) were the most frequent alleged methods, followed by suffocation, deprivation methods, forced body position, undressing and physical exercises (percentage between 49 percent and 29 percent). The frequency of suffocation, electricity, visual input reduced and threats was lower in 2000-2005 than in the 1992-1993 period. Different patterns of torture related to each police force were detected. The group arrested by the Guardia Civil alleged more severe torture methods, while the detainees arrested by Ertzantza alleged less severe ill-treatment. The prevalence of sexual torture was higher for women than for men. The present data are in consonance with the findings described for international organisms after their visits to Spain. INTERPRETATION: These findings, in addition to other evidence, suggest that torture is still a serious problem in Spain in relation with Basque incommunicado detainees. This fact shows that national and international (mainly based on CPT visits) measures of control/prevention have failed. This study supports the importance of scientific statistical analysis in the documentation of human rights violations and its potential use in order to improve the forensic evaluation of torture victims. PMID- 19289886 TI - Medical students' attitudes toward torture. AB - Torture, whether it be domestic or war related, is a public health issue of current concern. It is the position of the American Medical Association (AMA), The World Medical Association (WMA), the United Nations Declaration and the Geneva Convention, that torture is unethical, "morally wrong" and never to be condoned. The attitudes of medical students, our future physicians, will be critical in reducing the incidence of torture. The purpose of this investigation was to assess medical students' attitudes regarding the permissibility and ethics of the use of torture. A University of Illinois at Chicago College of Medicine's Institutional Review Board approved torture questionnaire was administered to 336 students of the University of Illinois College of Medicine. 35 percent of students agreed that torture could be "condoned" under some circumstances. Moreover, 24 percent of respondents disagreed that torture should "be prohibited" as a matter of state policy and a similar 24 percent disagreed that torture was "intrinsically wrong." It is concluded that most students felt that torture was "not permissible" and "intrinsically wrong", a disturbing 27 percent-35 percent felt that it could be permitted or condoned at times. Moreover, 27 percent felt that torture was not unethical. Given the strong condemnation of torture by the AMA, the WMA and the Geneva Convention these medical student attitudes, albeit by a minority of students, are disturbing. It is suggested that medical school curriculum committees review this matter. PMID- 19289887 TI - Pharmacological treatment to prevent and treat post-traumatic stress disorder. AB - Pharmacological treatments do have a role to play in the treatment of PTSD. Several agents have been shown to be superior to a placebo and many PTSD sufferers do appear to benefit from medication. The overall effect sizes are relatively small. It is to be hoped that in the future better pharmacological agents will be developed. INTRODUCTION: In recent years there has been a large increase in the amount of research looking at the neurobiology of post-traumatic stress disorder (PTSD). We now know that certain areas of the brain become active at the time of trauma, resulting in emotional and behavioural responses, and changes in neurochemicals and hormones. The amygdala, for example, is involved in the normal fear response, determines the significance of external stimuli and triggers responses such as fight, flight and freezing. These responses lead to alterations in stress hormones, neurochemicals and activity in other parts of the brain, such as the hippocampus and medial prefrontal cortex. One hypothesis is that in PTSD there is a failure of other networks to regulate amygdala reactivity, resulting in hyper reactivity to threat commonly seen in PTSD sufferers.1 Some, but not all studies have suggested that cortisol levels are lower in PTSD sufferers than in individuals without PTSD,2 and that there is adrenergic overactivity shortly after traumatic events. Our current knowledge of the neurobiology of PTSD, although not complete, suggests that certain drugs should be able to prevent its development and reduce its symptoms. PMID- 19289889 TI - Secondary trauma in the legal professions, a clinical perspective. AB - This article describes the importance of being aware of secondary trauma in lawyers, interpreters, judges, police, forensic physicians and other professionals that work with traumatized clients. In addition, it presents a psycho-educational model developed by the author to address secondary trauma among those associated with the legal and clinical professions. PMID- 19289890 TI - Justification doctrine in the prohibition on torture, cruel, inhuman or degrading treatment. AB - This paper looks at the legacy of a justification doctrine evident in early international jurisprudence that set the threshold for treatment prohibited under international law as torture, cruel, inhuman or degrading treatment or punishment [hereafter the prohibition], excusing from its reach deliberately inflicted, potentially severe, suffering proportionately inflicted for a legitimate purpose. Debates over a 'threshold' at which point the prohibition engages, or at which point 'inhumane' treatment reaches a level sufficient to be deemed 'torture', typically invoke an implicit 'severity' threshold. This paper is not primarily concerned with severity or the instrumentality of any 'severity threshold' either in engaging the prohibition or in distinguishing categories of prohibited treatment. Neither is the article concerned directly with the legal distinction between categories of prohibited treatment (i.e. the distinction between 'torture', other 'inhuman', or even any subcategory 'degrading' treatment). Rather the article focuses on the distinction between (i) treatment prohibited, as either torture or other cruel, inhuman or degrading, and (ii) treatment prima facie 'justified'. What the article looks at is the operation of a 'justification' threshold in triggering the prohibition, one that understands 'justified' treatment as never reaching the level of, or never amounting to inhuman, cruel or degrading treatment under the prohibition. The article interprets the current prohibition on torture, cruel, inhuman or degrading treatment as one on 'unjustified' inflicted suffering, suggesting that the notion of 'justifiability' active in this definition is problematic in encouraging arguments seeking to circumvent the protection afforded under the prohibition. In the absence of a clearly defined notion of the 'victim', or circumscribed class afforded protection, this paper both identifies and addresses a correlation between (i) a broadly inclusive contextual scope for the prohibition's applicability - one that contemplates a broad notion of the potential victim - and (ii) an enhanced role for a justification doctrine in excusing the infliction of [potentially severe] suffering where necessary and proportionate. In light of identified dangers associated with a role for justification doctrine in the definition of prohibited treatment, an alternative is put forward that would redefine the prohibition as one, not on 'unjustified' but one on 'all' suffering deliberately inflicted restricted to contexts of detention, custody, control or other deprivation of liberty. A brief disclaimer and clarification should also be made at the outset: The article addresses balancing exercises, active in determining the justifiability of treatment, that draw on the nature of its purpose and the degree of its severity. However the author wishes to make clear that the article in no way means to suggest that proportionality is, or should ever be, active in excusing treatment deemed cruel, inhuman, degrading or even torture; the article does not, in referring to 'balancing exercises', 'justification' or 'proportionality', mean to invoke, and much less to argue for, any justification doctrine or proportionality that would balance the prohibition against, for example, national security concerns. What the article is concerned with is a degree of balancing between the severity of suffering inflicted and a potentially legitimate purpose, operating in certain circumstances either to determine treatment as prohibited as torture, cruel, inhuman or degrading or alternatively to excuse it as 'justified'. It is not, then, balancing exercises which might mitigate (notwithstanding the absolute nature of the prohibition) the infliction of treatment deemed 'cruel, inhuman or degrading', or even that amounting to 'torture', but those balancing exercises which 'precondition' the triggering of the prohibition that are the subject of the article and of which will be attempted as lucid an analysis as possible. It is in this context that any reference to 'proportionality' in the article is made. Lastly the author wishes to clarify that anything presented or put forward by the article is done so solely in the interest of securing the maximum protection for the most vulnerable. PMID- 19289891 TI - Producing medico-legal evidence: documentation of torture versus the Saudi Arabian state of denial. AB - Five British men were detained and allegedly tortured in Saudi Arabia in 2000 and 2001. Two were sentenced to death and three were sentenced to 12 to 18 years of imprisonment. They received clemency in 2003 and were deported to England. After their homecoming, the men have unsuccessfully attempted to sue Saudi Arabian officials who had been responsible for their false accusations and human rights abuses. Saudi Arabia denies any form of torture and ill treatment of the five men, who maintain their allegations against the authorities responsible in Saudi Arabia. This article provides medical documentation of torture of the five British men by comparing: 1. Alleged torture methods, 2. Histories of immediate effects of torture, 3. Objective medical observations using clinical examinations as well as ultrasound scans of the victims' feet. The article concludes that there is a high degree of consistency between the claims of torture and observations, despite Saudi Arabian denial. PMID- 19289892 TI - Documentation of torture victims, assessment of the start procedure for medico legal documentation. AB - A Pilot Study was performed at the Rehabilitation and Research Centre for Torture Victims (RCT) in Copenhagen in order to explore the possibilities for adding a medico-legal documentation component to the rehabilitation of torture victims already taking place. It describes the process and results on implementing medico legal documentation in a rehabilitative setting. A modified version of the Guidelines in the Istanbul Protocol was developed on the basis of the review of literature and current practices described in "Documentation of torture victims, implementation of medico-legal protocols". The modified guidelines were tested on five clients. The aim was twofold: 1) To assess the client's attitude towards the idea of adding a documentation component to the rehabilitation process and: 2) To assess the practical circumstances of implementing the Istanbul Protocol in the everyday life of a rehabilitation centre. Results show that all five clients were positive towards the project and found comfort in being able to contribute to the fight against impunity. Also, the Pilot Study demonstrated that a large part of the medico-legal documentation was already obtained in the rehabilitation process. It was however not accessible due to lack of systematization and a data registering system. There are thus important synergies in collecting data for rehabilitation and documentation but a joint database system is necessary to realize these synergies. PMID- 19289893 TI - How therapists cope with clients' traumatic experiences. AB - An initial finding of high emotional stress in trauma therapists working in a specialized trauma institute led to three empirical studies on trauma-related therapist reactions. The purpose of these studies was to investigate the relation between high emotional burden and burnout, and the trauma-specific processes described by the concepts "secondary traumatic stress", "vicarious traumatization" and "traumatic countertransference". The initial qualitative/quantitative study examined how a group of specialized trauma therapists (N=63) coped with clients' traumatic experiences. The results on trauma-related reactions were inconclusive. This motivated a qualitative study of expert psychotherapists (N=11). Interviews with expert trauma therapists and other expert therapists were focused on reactions to the confrontation with traumatic experiences and differences between both groups of experts. Results indicated a specific reaction pattern to traumatic situations, but revealed no other differences between trauma specialists and other experts. To further examine trauma-specificity of this reaction pattern, a third study was conducted with psychology students (N=100) using an experimental design. The results suggest the existence of a trauma-specific reaction pattern, characterized by shock, anxiety and the experience of being carried away by strong emotions. The relation of trauma reactions with traumatic situations is endorsed by results on differential reactions to traumatic and interactionally difficult situations, although results suggest that other kinds of situations with high emotional impact may also evoke trauma-reactions. In the discussion the results are considered in relation to the limitations of the studies are followed recommendations for further research. Our results thus far support the high emotional impact of confrontation with traumatic material, but nuances psychopathological or other long-term negative changes that are suggested by the terms secondary or vicarious traumatization. PMID- 19289894 TI - Human rights abuses, transparency, impunity and the Web. AB - This paper reviews how human rights advocates during the "war-on-terror" have found new ways to use the World Wide Web (Web) to combat human rights abuses. These include posting of human rights reports; creating large, open-access and updated archives of government documents and other data, tracking CIA rendition flights and maintaining blogs, e-zines, list-serves and news services that rapidly distribute information between journalists, scholars and human rights advocates. The Web is a powerful communication tool for human rights advocates. It is international, instantaneous, and accessible for uploading, archiving, locating and downloading information. For its human rights potential to be fully realized, international law must be strengthened to promote the declassification of government documents, as is done by various freedom of information acts. It is too early to assess the final impact of the Web on human rights abuses in the "war-on-terror". Wide dissemination of government documents and human rights advocates' reports has put the United States government on the defensive and some of its policies have changed in response to public pressure. Even so, the essential elements of secret prisons, detention without charges or trials, and illegal rendition remain intact. PMID- 19289895 TI - Beyond where it started: a look at the "Healing Images" experience. AB - In March 2004, the Baltimore-based nonprofit organization Advocates for Survivors of Torture and Trauma (ASTT) initiated a photography-based therapeutic programme for clients. Developed by a professional photographer/teacher in collaboration with a psychologist, the programme has the goal of enabling clients to engage in creative self-exploration within a supportive, group setting. Since its inception, thirty survivors of conflict-related trauma and torture from five different countries have taken part in the programme, known as "Healing Images", using digital cameras to gather individually-chosen images that are subsequently shared and discussed within the group. These images include depictions of the natural and manmade environments in which clients find themselves; people, places and objects that offer comfort; and self-portraits that reflect the reality of the life of a refugee in the United States. This description of the "Healing Images" programme is based on comments gathered through discussion with participants and through interviews. Additional information was gathered from observation of early workshop sessions, review of numerous client photographs and captions, and pertinent organizational materials. A fundamental benefit of the programme was that it offered a mutually supportive group environment that diminished clients' feelings of psychological and physical isolation. Participants gained deep satisfaction from learning the technical skills related to use of the cameras, from the empowering experience of framing and creating specific images, and from exploring the personal significance of these images. Programme activities sparked a process of self-expression that participants valued on the level of personal discovery and growth. Some clients also welcomed opportunities to share their work publicly, as a means of raising awareness of the experience of survivors. PMID- 19289896 TI - The proposal for supervision training in Palestine/Middle East. AB - Clinical supervision has been known for decades. However, only in the past decade have training courses for supervision been developed and offered for health professionals working with a severely traumatized client population. Health professionals working as supervisors in this field are faced with specific problems. Together with Johan Lansen and Ton Haans, the Berlin Treatment Center for Torture Victims (bzfo) adapted the training method developed by Lansen/Haans in 1999, which offers participants a structural framework. In cooperation with the German Society for Supervision (DGSv), the bzfo offers this training course in Germany where it takes one and a half years to complete. To meet the vast need for supervision in countries where health professionals work under difficult and adverse conditions, the bzfo is now in contact with colleagues in the Gaza region with a proposal for supervision tailored to their specific needs. PMID- 19289897 TI - The role of the Istanbul-Protocol in the uphill battle for torture survivors being granted asylum in Europe and ensuring the perpetrators pay. PMID- 19289898 TI - [Associations of emotional state and quality of life with lipid concentration, duration of the disease, and the way of treating the disease in persons with type 2 diabetes mellitus]. AB - The aim of the study was to evaluate associations of emotional state and quality of life with lipid concentration, duration of the disease, and the way of treating the disease in males and females with type 2 diabetes mellitus. A total of 53 persons with type 2 diabetes mellitus (27 males and 26 females; mean age, 58.7+/-8.9 years) and 56 healthy persons (26 males and 30 females; mean age, 54.7+/-8.3 years) participated in the study. Emotional state was evaluated by means of Profile of Mood State and quality of life by means of WHO Brief Quality of Life Questionnaire. Emotional state and quality of life were significantly worse, tension-anxiety and fatigue-inertia were significantly higher, vigor activity was significantly lower in male patients with type 2 diabetes mellitus than in healthy males. In females, no significant differences in emotional state and quality of life comparing type 2 diabetes mellitus group and controls were detected. In females with type 2 diabetes mellitus, emotional state and quality of life were significantly better, scores of tension-anxiety, depression dejection, anger-hostility, and fatigue-inertia were significantly lower, and score of vigor-activity was significantly higher than in males with type 2 diabetes mellitus. Some significant correlations were found. In males, vigor activity correlated with total cholesterol level and negatively correlated with triglyceride level. In females, significant correlations were found between scores of emotional state (tension-anxiety, depression-dejection, confusion bewilderment, and total score of emotional state) and lipid levels (total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels). There were no significant associations of emotional state and quality of life with duration of the disease in males and females with type 2 diabetes mellitus. No significant differences in emotional state and quality of life were found between males and females with type 2 diabetes mellitus, who were treated with oral antidiabetic preparations and insulin preparations. PMID- 19289899 TI - Significance of dietotherapy on the clinical course of atopic dermatitis. AB - The aim of this study was to determine the efficiency of individual balanced replacement diet in treatment of children with atopic dermatitis, to compare the course of atopic dermatitis and gastrointestinal disorders, as well as the data of skin patch test after a one-year period of dietary treatment. PATIENTS AND METHODS: The study group included 154 children (their age varied from 6 months to 18 years) with atopic dermatitis, for whom food allergens were determined by allergic skin tests (skin prick and patch). These children were recommended an individual balanced replacement diet, where possible food allergens were replaced by other products that do not cause allergic reactions. After a one-year dietary treatment, 109 (70.8%) children (such number came for the second study) were tested repeatedly. The following aspects were evaluated for all these children: clinical course of atopic dermatitis (children's mothers provided answers about exacerbation of allergic rash during the last 12 months, gastrointestinal disorders, and used medicines), severity of the progress of atopic dermatitis (SCORAD index). Besides, skin patch test with 25 food allergens was carried out. RESULTS: Children who followed dietary recommendations were younger than children who failed to follow dietary recommendations because of a variety of reasons (P=0.01). Even 49 (62.8%) patients who followed dietary recommendations have shown the following results during the second test: allergic rash disappeared and they did not have to take medicines against allergy anymore. Patients who followed their individual dietary recommendations more rarely suffered from severe allergic rash problems during a 12-month period (P=0.01) and they had to take fewer medicines against allergy, compared to children who did not follow their dietary recommendations (P=0.001). Clinical course of atopic dermatitis in children who followed individual dietary recommendations was easier compared to children who did not follow such recommendations (P=0.001). During a one-year dietary treatment, 28.2% of children with atopic dermatitis became more tolerant to earlier food allergens. After the comparison of skin patch test results (before dietary treatment and after a one-year period), it was determined that only skin patch tests against buckwheat, oat, beef, and cacao did not change statistically significantly. Results of skin patch tests against other food products were found to be positive more rarely. Besides, children who followed their dietary recommendations suffered from gastrointestinal disorders more rarely as compared to children who did not follow their dietary recommendations (P=0.01). They suffered less from abdominal pain (P=0.01), abdominal distention (P=0.044), and constipation (P=0.035). CONCLUSIONS: Individual balanced replacement diet for children with atopic dermatitis helped to fully control nutrition of sick children from various age groups and had a positive effect on the clinical course of atopic dermatitis. Patients who followed their individual dietary recommendations suffered from severe allergic rash more rarely and they had to take fewer medicines against allergy as compared to children who did not follow dietary recommendations. Clinical course of atopic dermatitis in children who followed individual dietary recommendations was easier as compared to children who did not follow such recommendations. One-third of children with atopic dermatitis became more tolerant to earlier food allergens during a one year period. After a one-year dietotherapy treatment, positive patch test reactions to many food products appeared to be more rarely, except for buckwheat, oat, beef, and cacao. Besides, children who followed their dietary recommendations suffered from gastrointestinal disorders (abdominal pain, abdominal distention, and constipation) significantly more rarely as compared to children who did not follow their dietary recommendations. PMID- 19289900 TI - Implementation of international transtelephonic ECG platform for patients with ischemic heart disease. AB - Ischemic heart disease in European countries over decades causes up to 55% of all cases of sudden death and also has a high rate of mortality, morbidity, and hospital admission. Patients with such chronic diseases also require intensive home care facilities from community nurses. The aim was to establish international multilingual platform for transtelephonic ECG system as an alternative solution for home care and assess its performance. METHODS. During this pilot study, the international toll-free line between Lithuania and Germany was established, and practical applicability of the tele-ECG device was tested. Transtelephonic ECG system was implemented between Telemedicine Center in Bad Segeberg (Bad Segeberg Clinic, Germany), the Call Center in Kaunas at the Hospital of Kaunas University of Medicine, and a patient residence. RESULTS. Over a 6-month follow-up period, 34 patients were recruited. Following the ECG transmission, 86 teleconsultations were done. During the study, a total of 329 ECGs were sent by the patients; out of them, 14 ECGs were with clinical changes. Technical problems due to insufficient patient training, telecommunication systems, acoustic data transmission, and device itself were reported. Up to 23% of ECGs sent by patients were unreadable and not applicable for further clinical analysis. CONCLUSIONS. Our study showed the potential of telemedicine facilities to overcome the problems of access that makes the technique so potentially useful, but for telemonitoring application at patient homes in a wider population, it needs to be improved in terms of technical performance, transmission and analysis automatization. PMID- 19289901 TI - The measurements of health-related quality-of-life and pain assessment in the preoperative patients with low back pain. AB - OBJECTIVE: This prospective observational study of the Short-Form Health Survey (SF-36), Oswestry Disability Index, Lithuanian version of the McGill Pain Questionnaire, and Visual Analogue Scale (VAS) for pain was performed to evaluate their effectiveness in the additional preoperative screening of patients with disc herniation disease. PATIENTS AND METHODS: In the present study, we investigated a cohort of 100 patients with lumbar disc herniation causing low back pain and the second one of 100 patients with nonspecific low back pain by applying physical activity, pain scales and Short-Form 36 General Health Questionnaire. RESULTS: The quantitative analysis of SF-36 domain scores showed the substantial differences in both examined (herniated and control) groups. In the present study, we estimated moderate but statistically significant (P<0.05) correlations between the bodily pain domain scores and assessment of back and leg pain on the VAS, as well as between the physical function and walking/standing ability (Oswestry). According to appropriate pain assessment instruments (Lithuanian version of the McGill Pain Questionnaire), qualitative and quantitative analysis of the preoperative patients was performed. CONCLUSION: The provided methodology could be used in population-based studies or in clinical samples that focus on specific impairments and seek to control the pain frequency and intensity, for example, follow-up assessments testing the effectiveness of surgical procedures performed, and to elicit the pathways leading to other impairments. PMID- 19289902 TI - Histology of human glioblastoma transplanted on chicken chorioallantoic membrane. AB - Glioblastoma is the most malignant tumor in the range of cerebral astrocytic gliomas. A lot of experimental models are used to evaluate various properties of glioblastoma. Chicken chorioallantoic membrane model is one of them. OBJECTIVE. To evaluate histology and survival of glioblastoma tumors taken immediately from operating theatre and transplanted on chicken chorioallantoic membrane. MATERIALS AND METHODS. Glioblastoma samples obtained from 10 patients were transplanted onto 200 eggs. Overall, we used 15 tumors; only 5 of them were not glioblastomas as it was revealed later. RESULTS. The transplanted tumors survive up to 6 days. Transplants do not survive longer because during embryo's development the nourishing membrane dries. Transplanted glioblastomas exhibited the same features as original glioblastomas - necrosis, endothelium proliferation, cellular polymorphism - while transplanted glioblastomas also showed glial fibrillary acidic protein (GFAP), vimentin, Ki67, S100 protein, neurofilament immunoreactivity, and infiltration of macrophages (CD68) and T cells (CD3(+), CD8(+)). Transplanted glioblastomas did not show any immunoreactivity of p53. Invasion of vessels from the chicken into transplanted tumor is not observed. Chicken erythrocytes did not appear within the transplants, and tumor cells invade chicken tissue at the minimum. CONCLUSION. Our data show that transplanted pieces of glioblastoma survive with all cytological features. The presence of macrophages (marker CD68) and T cells (markers CD3(+) and CD8(+)) can be registered in the transplant. The data revealed that transplanted glioblastoma remains as insulated unit, which survives from nourishment of the chorioallantoic membrane apparently only by diffusion. The features of original tumor-host reaction of the patient remained too. PMID- 19289903 TI - [The types of deliberate self-harm and its prevalence among Lithuanian teenagers]. AB - The aim of this study is to identify how widely deliberate self-harm is spread and the types of deliberate self-harm among 15-17-year-old teenagers in Lithuania. MATERIAL AND METHODS: The anonymous Lifestyle and Coping Questionnaire was used for the study carried out in 2006. This countrywide study involved 3848 respondents (2200 girls (57.2 %) and 1648 boys (42.8%)) aged 15-17 years from all 10 regions of Lithuania. RESULTS: According to the findings of this study, 7.3% of 15-17-year-old Lithuanian teenagers (9.9% girls and 3.8% boys) stated that they had deliberately overdosed drugs or tried to inflict self-injury in other ways. Less than half of them (43.4%) reported that they were thinking to repeat such behavior. Half of the adolescents who inflicted self-injury were living in a two-parent family, 27.4% - with one of the parents, 7% - with other member of the family, and 9% - with other people. One-third of adolescents (34.2%) choose internal ways of self-harm, 26% - external self-injury, 11% - tried to harm themselves in mixed way, and 28.8% - did not indicate the way. In case of a self injury mentioned above, 13.5% of adolescents were admitted to hospital. CONCLUSIONS: In Lithuania, adolescent self-harm is relatively frequent: 7.3% of 15-17-year-old Lithuanian adolescents deliberately self-harmed during their life. The most frequent way to self-harm is to overdose. This study confirms the need for preventive activities and necessity of further studies in this field. PMID- 19289904 TI - [Prevalence of conventional risk factors in patients with coronary heart disease]. AB - Extensive clinical and statistical studies have identified risk factors that increase the incidence of coronary heart disease. It is commonly suggested that more than 50% of patients with coronary heart disease lack any of the conventional risk factors. OBJECTIVE. To determine the prevalence of four conventional risk factors among patients with coronary heart disease. MATERIAL AND METHODS. We analyzed data of 606 patients with coronary heart disease (myocardial infarction, unstable and stable angina pectoris) hospitalized in the Clinics of Santariskes, Vilnius University Hospital, in 1997-2005. RESULTS. Among patients with coronary heart disease, at least one of four conventional risk factors was present in 98% of patients. Hypertension was present in 47.7% of patients, diabetes - in 12.9%, dyslipidemia - 90.1%, and smoking - in 24.1% of patients. In younger patients (<55 years), only 2.3% of patients lacked any of four conventional risk factors. Two-thirds (66.5%) of younger patients with coronary heart disease had two and more risk factors. CONCLUSIONS. Considering the fact that patients with coronary heart disease often lack conventional risk factors, currently more attention is given to nontraditional risk factors as well as genetic causes of coronary heart disease. Nevertheless, the present study revealed that 98% of patients with coronary heart disease had at least one of four conventional risk factors. Among younger patients (younger than 55 years), conventional risk factors are identified very frequently. Thus, it can be concluded that in order to reduce the epidemic of coronary heart disease, much greater emphasis should be given to identify and to improve prevention of four conventional risk factors as well as the lifestyle of the patient. PMID- 19289905 TI - The prevalence of malocclusion among 7-15-year-old Lithuanian schoolchildren. AB - The epidemiological data on the prevalence of malocclusion is an important determinant in planning appropriate levels of orthodontic services. The occurrence of occlusal anomalies varies between different countries, ethnic and age groups. The aim of this study was to describe the prevalence of malocclusion among Lithuanian schoolchildren in the 7-9-, 10-12-, and 13-15-year age groups assessing occlusal morphology. The study included 1681 schoolchildren aged 7-15 years. The crowding, spacing, overbite, overjet, the relationship of the first upper and lower molars according Angle's classification, and posterior crossbite were assessed. The study demonstrated that only 257 children had normal occlusion, and 44 had undergone orthodontic treatment among them. The greatest overjet in the studied contingent was 11 mm, and the negative overjet - 3 mm. The overbite ranged between 0 and 6 mm with a mean of 2.29+/-1.23 mm. Posterior crossbite was recorded in 148 children (8.8%). This study showed that the prevalence of malocclusion among 7-15-year-old Lithuanian schoolchildren is 84.6%. The most common malocclusion was dental crowding. The upper dental arch crowding was registered for 44.1% and lower for 40.3% of all schoolchildren. The class I molar relationship was detected in 68.4% of the subjects, class II - in 27.7%, and class III - in 2.8%. PMID- 19289906 TI - Health of Roma children in Vilnius and Ventspils. AB - According to the literature data, Roma health and living conditions in Central and Eastern Europe are poorer than of the rest of population. However, the more detailed information about Roma health is lacking. The aim of the study was to evaluate morbidity, health self-assessment, and prevalence of addictions among Roma children in Vilnius and Ventspils and to compare with health indicators of non-Roma children. PARTICIPANTS AND METHODS. A descriptive epidemiological study was carried out, anonymously questioning all volunteer Roma children - 59 in Vilnius (Lithuania) and 31 in Ventspils (Latvia) schools. Results were compared with identical study, carried out in five Vilnius schools (reference group, 640). RESULTS. The appliance rate of Roma children to doctor did not differ from reference group - half of all questioned children visited doctor 1-3 times during the last year. However, more Roma children (74.6% from Vilnius and 64.6% from Ventspils) considered their health as poor and very poor as compared to reference group (4.3%). The proportion of children indicating somatic symptoms often and very often did not differ statistically significantly among groups with exception of vomiting and nausea, which was most prevalent among Vilnius Roma and Ventspils Roma. The proportion of children indicating emotional symptoms often and very often differed significantly in all groups and was the biggest in Ventspils Roma group. The proportion of daily alcohol, drug users, and smokers was higher in Vilnius Roma and Ventspils Roma groups, although the differences among all three groups were not statistically significant. CONCLUSION. Although the morbidity of Vilnius Roma and Ventspils Roma groups did not differ from reference group, essential discrepancy was found in health self-assessment - more Roma children considered their health as poor and very poor. PMID- 19289907 TI - [Tumor and immunity]. AB - System of organism defense is an important complex of interrelated cellular, molecular, genetic, and other components, which regulate homeostasis of host. Experimental and clinical data show that immune system functions are significant, but also a complicated question in cancer development. It is very important to investigate and understood how immune system coordinates the response to cancer cells. Our understanding about innate and adaptive immunity functions and interaction with transformed cells is constantly changing. Different responses of these system components can promote, reduce, or inhibit tumor development. It is established that malignant cells develop into invasive cancer with interaction with tumor microenvironment, which is influenced by inflammation. Clinical and experimental studies have revealed the link between inflammation and cancer risk. Many cancers develop in the sites of inflammation. Activation of humoral and cellular immunity may predispose to neoplastic or cancer development. Despite the scientific progress, understanding of the immune system mechanisms responding to malignance remains insufficient. PMID- 19289908 TI - From the Editor-in-Chief: leadership for the information age: the time for action is now. AB - Dr. Lynn Nagle, the Senior Nursing Advisor for Canada Health Infoway, writes the column on nursing informatics for this journal. She and I have both been involved in the development and now the implementation of HOBIC (Health Outcomes for Better Information and Care), a province-wide initiative funded by the Ontario Ministry of Health and Long-Term Care: I am the executive lead and Lynn is the informatics lead. HOBIC seeks to bring online functionality to nurses that supports systematic assessment of patients on eight outcomes upon admission and discharge in acute care, chronic hospital care, long-term care and home care, and quarterly for people in residential settings. There is strong research evidence that nurses make a difference in how well patients do on these outcomes. Nurses can now access the results of their assessments online throughout a patient's stay, compare them to other patients of similar age or gender and begin to set benchmarks for improving these outcomes. Unit managers and chief nursing officers receive an array of monthly reports on the admission and discharge status of patients - information that can also be reviewed on the basis of gender and age group. HOBIC will be a critical component of the electronic health record when it is wholly adopted throughout Ontario. PMID- 19289909 TI - Times of crisis, times of action. AB - At the recent Academy of Canadian Executive Nurses (ACEN) Annual Meeting in November 2008 in Ottawa, I had the privilege and honour of becoming president of this dynamic organization. Founded more than 30 years ago, ACEN represents the voice of senior academic nursing leadership in Canada, helping to set directions for healthcare policy and contributing to the alignment and advancement of the national agendas for nursing practice, education, research and leadership. PMID- 19289910 TI - Global nurse migration: its impact on developing countries and prospects for the future. AB - The worldwide shortage of nurses, which results from a global undersupply and high attrition rates, affects developed countries in the West the same way as it affects developing countries in Asia, Africa and Latin America. The difference lies in the fact that developing countries serve as a readily available source of trained nurses for developed countries in Europe, North America and parts of Oceania. Strong "pull" and "push" factors favour wealthier nations in the West in their efforts to deal with domestic shortages through overseas recruitment. Thus, the ongoing nursing shortage in developing countries is worsened by a loss of thousands of trained nurses every year to emigration. This paper brings into focus the magnitude of the problem in terms of the number of nurses migrating to and from various countries and its impact on developing countries. The paper also examines some of the ongoing efforts in developing countries to mitigate the problem, and sheds light on the prospects for improvement in the foreseeable future. PMID- 19289911 TI - We are called upon to lead. AB - The paper "Global Nurse Migration: Its Impact on Developing Countries and Prospects for the Future" by Khaliq, Broyles and Mwachofi explores the global nurse migration experience in developed and developing countries. Nurses choose to migrate from developing countries in response to economic and political instability, and poor salaries and working conditions in their home countries. These factors also interfere with the home countries' capacity to deliver education for health professions and to provide adequate support for a healthcare system. The paper clearly outlines the catastrophic outcomes of nurse migration for developing countries where inadequate numbers of trained and experienced nurses remain to support education and administration. Many well-documented references in the paper report overburdening of the remaining workforce, reduced access to care, increased risk of patient mortality, increased maternal, infant and under-five-year mortality rates, reduced life expectancy, increased incidence of communicable diseases and reduced immunization, all resulting from shortages of health professionals, leading to increased nursing workload and burnout. PMID- 19289912 TI - Canadian strategies to stabilize the work force. AB - Today's globalized economy creates opportunities for health professionals but corresponding challenges for countries facing significant shortages of these professionals. The uneven playing field between developed and developing countries hampers the latter in recruiting and retaining trained professionals to oversee and maintain their health systems. Given the salary differentials and variance in working conditions between developed and developing countries, developing countries may lack the pull factors to keep their nurses and doctors from emigrating. However, many developed countries have made significant investments to address this challenge. PMID- 19289913 TI - Aligning career development with organizational goals: working towards the development of a strong and sustainable workforce. AB - The rapidly changing world of healthcare is faced with many challenges, not the least of which is a diminishing workforce. Healthcare organizations must develop multiple strategies, not only to attract and retain employees, but also to ensure that workers are prepared for continuous change in the workplace, are working at their full scope of practice and are committed to, and accountable for, the provision of high-quality care. There is evidence that by creating a healthier workplace, improved patient care will follow. Aligning Healthy Workplace Initiatives with an organization's strategic goals, corporate culture and vision reinforces their importance within the organization. In this paper, we describe an innovative pilot to assess a career development program, one of multiple Healthy Workplace Initiatives taking place at Providence Care in Kingston, Ontario in support of our three strategic goals. The results of the pilot were very encouraging; subsequent success in obtaining funding from HealthForceOntario has allowed the implementation of a sustainable program of career development within the organization. More work is required to evaluate its long-term effectiveness. PMID- 19289914 TI - The effects of emotionally intelligent leadership behaviour on emergency staff nurses' workplace empowerment and organizational commitment. AB - The purpose of this study was to test a model exploring the relationships among emotionally intelligent leadership behaviour, workplace empowerment and commitment. A predictive, non-experimental design was used to test the model in a random sample of 300 emergency staff nurses working in Ontario. A path analysis supported the fully mediated hypothesized model (chi(2)=2.3, df=1, p > .05; CFI=.99, IFI=.99, RMSEA=.08). Perceived emotionally intelligent leadership behaviour had a strong direct effect on structural empowerment (beta=.54), which in turn had a strong direct effect on organizational commitment (beta=.61). PMID- 19289915 TI - Nursing leaders' accountability to narrow the safety chasm: insights and implications from the collective evidence base on healthcare safety. AB - Challenges continue to exist in bridging the safety gap to ensure that consistent, high-quality nursing care is provided based on the best scientific knowledge available. This paper examines findings from nursing research presented at the symposium Advancing Nursing Leadership for a Safer Healthcare System, held in Toronto, Ontario in 2007. Four central themes emerged: (1) place the patient in safety; (2) generate a broader knowledge base on safety across the continuum of care; (3) create a safe culture and healthy work environment to mitigate current threats to patient safety; and (4) advance translation of evidence to practice at the organizational and clinical levels. The aim of this exchange of knowledge was to equip nursing leaders and their decision partners with evidence that can become a catalyst for mobilizing change in practice to address the safety chasm. PMID- 19289916 TI - Infusion teams: a critical element of patient care. PMID- 19289918 TI - Public policy mandating needlestick injury prevention generates research and change creating safer workplaces. PMID- 19289919 TI - Reducing the risk of air embolism. PMID- 19289920 TI - Pharmacological interventions to treat phlebitis: systematic review. AB - This study presents a systematic review for evaluating effective pharmacological actions for the treatment of phlebitis stemming from infusion therapy. The studies reviewed were categorized according to the type of therapeutic approach proposed by the author and by the level of evidence presented. The review found that topical nitroglycerin and notoginseny were more effective in the reduction of the inflammatory process when compared with other proposed alternatives. Nevertheless, the development of research related to possible alternatives for the treatment of phlebitis is important. PMID- 19289921 TI - A broad-spectrum look at catheter-related bloodstream infections: many aspects, many populations. AB - The issue of central vascular catheter-related bloodstream infections is prominent in medical literature and practice today, both in the United States and throughout the international healthcare community. In its 2002 report on Guidelines for the Prevention of Intravascular Catheter-Related Infections, the Centers for Disease Control and Prevention estimated that 250,000 cases of central vascular catheter-related infections occur annually. This article is a review of current literature pertaining to catheter-related bloodstream infections. Included are resources from 2004 onward, as well as resources such as the Centers for Disease Control and Prevention, the Institute for Healthcare Improvement, and the National Guideline Clearinghouse. PMID- 19289922 TI - A review of primary hyperparathyroidism. AB - Primary hyperparathyroidism is the inappropriate secretion of parathyroid hormone and is the most common cause of hypercalcemia. Most patients with primary hyperparathyroidism are asymptomatic, and the disease is more common in postmenopausal women than in older men. Primary hyperparathyroidism can be treated with medication and long-term monitoring; however, surgical removal of the affected parathyroid gland is the definitive treatment. Rarely occurring, parathyroid crisis must be treated promptly to prevent deleterious effects to the patient. The patient experiencing parathyroid crisis benefits from the infusion nurse's knowledge of drug and fluid therapies and expertise in peripheral access. PMID- 19289923 TI - External jugular cannulation in infants and children. AB - Placement of a peripherally inserted central catheter (PICC) is commonplace in infants and children for the infusion of medications, hydration, and nutritional solutions. Vein depletion caused by repeated and prolonged need for vascular access devices has forced practitioners to consider alternate veins for providing care. The external jugular vein has a positive history of use for insertion of the PICC and is becoming increasing popular for this purpose. Pertinent anatomy, patient selection criteria, preparation, and catheter insertion and maintenance processes related to the catheter placed and residing in the external jugular vein are discussed. PMID- 19289925 TI - Adjuvant treatment with vindesine for metastatic melanoma: in response to the study of Eigentler et al. (2008). PMID- 19289926 TI - Blockade of glutamate transporters facilitates cerebellar synaptic long-term depression. AB - Excitatory amino acid transporters (EAATs) are believed to limit extracellular glutamate concentrations with specific roles poorly understood. At cerebellar climbing fiber-Purkinje cell synapse, EAAT4 and metabotropic glutamate receptor 1 (mGluR1) are closely expressed in surrounding postsynaptic locations, suggesting that EAAT4 may regulate mGluR1 activation. We examined the actions of EAAT4 on synaptic plasticity by applying blockers of glutamate transporters, DL-threo-beta benzyloxyaspartic acid and D-aspartate. Inhibition of EAAT4 markedly prolonged AMPA receptor-mediated excitatory postsynaptic currents evoked by stimulating climbing fibers. Impairing glutamate uptake facilitated mGluR1-dependent climbing fiber-Purkinje cell synaptic long-term depression (LTD). Glutamate uptake blockers also sufficiently rescued climbing fiber-Purkinje cell synaptic LTD that failed to be induced by a weaker tetanus. Our results suggest that neuronal glutamate transporters strongly influence mGluR1-dependent cerebellar LTD. PMID- 19289927 TI - Peer review and the nursing literature. PMID- 19289928 TI - Clinical trial of tailored activity and eating newsletters with older rural women. AB - BACKGROUND: Unhealthy diet and lack of physical activity increase rural midlife and older women's risk of chronic diseases and premature death, and they are behind urban residents in meeting Healthy People 2010 objectives. OBJECTIVES: The objective of this study was to compare a tailored intervention based on the Health Promotion Model with a generic intervention to increase physical activity and healthy eating among rural women. METHODS: In a randomized-by-site, community based, controlled, clinical trial, Wellness for Women, 225 women aged 50 to 69 years were recruited in two similar rural areas. Over 12 months, women received by mail either 18 generic newsletters or 18 newsletters computer tailored on Health Promotion Model behavior-specific cognitions (benefits, barriers, self efficacy, and interpersonal support), activity, and eating. Outcomes at 6 and 12 months included behavioral markers and biomarkers of physical activity and eating. Data were analyzed by repeated-measures analysis of variance and chi square tests (alpha < .05). RESULTS: Both groups significantly increased stretching and strengthening exercise and fruit and vegetable servings and decreased percentage of calories from fat, whereas only the tailored group increased moderate or greater intensity activity and decreased percentage of calories from saturated fat from baseline to 6 months. Both groups increased stretching and strengthening exercise, whereas only the tailored group increased moderate or greater intensity activity and fruit and vegetable servings and decreased percentage of calories from fat from baseline to 12 months. Both groups had several changes in biomarkers over the study. A higher proportion of women receiving tailored newsletters met Healthy People 2010 criteria for moderate or greater intensity activity, fruit and vegetable servings, and percentage of calories from fat at 12 months. DISCUSSION: Mailed computer-tailored and generic print newsletters facilitated the adoption of change in both activity and eating over 6 months. Tailored newsletters were more efficacious in facilitating change over 12 months. PMID- 19289929 TI - A national online forum on ethnic differences in cancer pain experience. AB - BACKGROUND: Cultural values and beliefs related to cancer and pain have been used to explain ethnic differences in cancer pain experience. Yet, very little is known about similarities and differences in cancer pain experience among different ethnic groups. OBJECTIVE: The objective of this study was to explore similarities and differences in cancer pain experience among four major ethnic groups in the United States. METHODS: A feminist approach by Hall and Stevens was used. This was a cross-sectional qualitative study among 22 White, 15 Hispanic, 11 African American, and 27 Asian patients with cancer recruited through both Internet and community settings. Four ethnic-specific online forums were conducted for 6 months. Nine topics related to cancer pain experience were used to guide the online forums. The collected data were analyzed using thematic analysis involving line-by-line coding, categorization, and thematic extraction. RESULTS: All participants across ethnic groups reported "communication breakdowns" with their healthcare providers and experienced "changes in perspectives." All of them reported that their cancer pain experience was "gendered experience." White patients focused on how to control their pain and treatment selection process, whereas ethnic minority patients tried to control pain by minimizing and normalizing it. White patients sought out diverse strategies of pain management; ethnic minority patients tried to maintain normal lives and use natural modalities for pain management. Finally, the cancer pain experience of White patients was highly individualistic and independent, whereas that of ethnic minority patients was family oriented. DISCUSSION: These findings suggest that nurses need to use culturally competent approaches to cancer pain management for different ethnic groups. Also, the findings suggest further in depth cultural studies on the pain experience of multiethnic groups of patients with cancer. PMID- 19289930 TI - Pressure ulcer preventive device use among elderly patients early in the hospital stay. AB - BACKGROUND: Clinical guidelines for the prevention of pressure ulcers advise that pressure-reducing devices should be used for all patients at risk of or with pressure ulcers and that all pressure ulcers should be documented in the patient record. Adherence to these guidelines among elderly hospital patients early in the hospital stay has not been examined in prior studies. OBJECTIVE: The objective of this study was to examine adherence to guidelines by determining the frequency and correlates of use of preventive devices early in the hospital stay of elderly patients and by determining the frequency and correlates of recording pressure ulcers in the patient record. METHODS: This was a cross-sectional study of 792 patients aged 65 years or older admitted through the emergency department to the inpatient medical service at two teaching hospitals in Philadelphia, Pennsylvania, between 1998 and 2001. Patients were examined by a research nurse on Hospital Day 3 (median of 48 hours after admission) to determine the use of preventive devices, presence of pressure ulcers, and risk of pressure ulcers (by Norton scale). Data on additional risk factors were obtained from the admission nursing assessment in the patient record. Data on documentation of pressure ulcers were obtained by chart abstraction. RESULTS: Only 15% of patients had any preventive devices in use at the time of the examination. Among patients considered at risk of pressure ulcers (Norton score < or =14), only 51% had a preventive device. In multivariable analyses, high risk of pressure ulcers was associated with use of preventive devices (odds ratio = 41.8, 95% confidence interval = 14.0-124.6), whereas the type and stage of pressure ulcer were not. Documentation of a pressure ulcer was present for only 68% of patients who had a pressure ulcer according to the research examination. DISCUSSION: Use of preventive devices and documentation of pressure ulcers are suboptimal even among patients at high risk. PMID- 19289931 TI - Significant impairment in immune recovery after cancer treatment. AB - BACKGROUND: Although immunosuppression from cancer adjuvant therapy has been documented, how these suppressed immune responses recover to baseline values after completion of cancer adjuvant therapy has not been studied systematically. OBJECTIVES: The objective of this study was to examine the probability of immune recovery after cancer adjuvant therapy and the potential impact of cancer adjuvant therapy type and cancer stage on immune recovery in patients with newly diagnosed breast cancer. METHODS: In a repeated-measures design, immune responses were measured four times in 80 patients with early-stage breast cancer: before and at 2, 6, and 12 months from the beginning of cancer adjuvant therapy. Natural killer cell activity, lymphokine-activated killer cell activity, lymphocyte proliferation, CD subsets (CD4, CD8, and CD56), and cytokines (interferon-gamma, interleukin [IL]-2, IL-4, IL-6, and IL-1alpha) were selected for their relevance to breast cancer. Immune recovery was defined by the level of immune response reaching to and above baseline levels. Data were analyzed using a multivariate generalized linear mixed-model approach. RESULTS: Delayed immune recovery to pretreatment baseline levels continued to the 12-month time point in all parameters. The percentages of immune recovery ranged from 6% to 76% of the patients, varying among immune parameters. Overall, immune recovery was poorer for interferon-gamma, IL-2, IL-4, lymphocyte proliferation, and natural killer cell activity than was for CD subsets and IL-6. The type of cancer adjuvant therapy, not cancer stage, showed selective influence on immune recovery. Chemotherapy or chemotherapy and radiotherapy combination significantly delayed IL-2 recovery, whereas radiotherapy significantly delayed IL-4 recovery. DISCUSSION: Immune recovery after breast cancer adjuvant therapy is delayed significantly for an extended time period in numerous immune parameters. The type of cancer adjuvant therapy has selective influence on immune recovery. Future investigations are warranted to elucidate the time course of immune recovery, clinical significance of poor immune recovery, and factors influencing immune recovery to develop potential interventions. PMID- 19289932 TI - Factors identified by experts to support decision making for post acute referral. AB - BACKGROUND: Each year, more than 13 million post acute referral decisions are made for Medicare recipients, yet there are no national, empirically derived decision support tools to assist in making these important decisions. OBJECTIVES: The aim of this study was to elicit expert knowledge about factors important to referral decision making and identify the characteristics of hospitalized patients who need a post acute referral. METHODS: This was a retrospective and prospective mixed-methods study of the referral decisions made by discharge planning experts for 355 hospitalized older adults. Variables included sociodemographics, living arrangement, insurance, diagnosis, comorbid conditions, adverse events, medications, home care use, hospitalization in last 30 days or 6 months, patients' perception of need for and use of assistive devices or post acute services, length of stay, cognition, self-rated health, depression, functional status, and post acute referral decision. RESULTS: The final model identified six factors associated with the need for a post acute referral. A cutpoint was derived with a sensitivity and specificity of 87.6% and 63.2%, respectively. Experts were more likely to refer patients who had no or intermittent help available (odds ratio [OR] = 3.0), major walking restrictions (OR = 6.5), less than excellent self-rated health (3.1 and 4.0 times more likely with good and fair-poor health, respectively), remained in the hospital longer (OR = 1.2), and had higher depression scores (OR = 1.1) or number of comorbidities (OR = 1.2). DISCUSSION: This study begins to identify information useful to clinicians caring for hospitalized older adults who may benefit from post acute services. By assuring the systematic, valid, and reliable collection of these items, the multidisciplinary team is alerted to patients who may benefit from post acute services. Further work is needed to increase the specificity and generalizability of the model and to test its effects on patient and clinician outcomes. PMID- 19289933 TI - Simpson's paradox and experimental research. AB - BACKGROUND: Experimental research in nursing has increased considerably in recent years. To improve the quality of such research, it is critical to reduce threats to internal validity. One threat that has received inadequate attention in the nursing literature is Simpson's paradox--a case of extreme confounding that can lead to erroneous conclusions about the effects of an experimental intervention. In fact, it can lead to a conclusion about an intervention effect that is the opposite of the correct inference. APPROACH: The aims of this study were to describe Simpson's paradox, provide a hypothetical example, and discuss approaches to avoiding the paradox. RESULTS: The paradox is due to the combination of an overlooked confounding variable and a disproportionate allocation of that variable among experimental groups. Different designs and analysis approaches that can be used to avoid the paradox are presented. DISCUSSION: Simpson's paradox can be avoided by selecting an appropriate experimental design and analysis that incorporates the confounding variable in such a way as to obtain unconfounded estimates of treatment effects, thus more accurately answering the research question. PMID- 19289934 TI - Planning a cluster randomized controlled trial: methodological issues. AB - BACKGROUND: The standard approach in a randomized controlled trial (RCT) is to randomize individuals to intervention and control groups. Yet, nursing and other health interventions are often implemented at the levels of health service organizational unit or geographical area. It may be more appropriate to conduct a cluster RCT. However, cluster randomization requires consideration of a number of important issues. OBJECTIVE: The objective of this study was to show how critical issues in relation to design and analysis can be addressed. APPROACH: Two cluster RCTs conducted by the authors are used as examples. Guidance on the conduct and reporting of cluster RCTs is also offered. RESULTS: A rationale for choosing this design was provided, and issues in relation to study design, calculation of sample size, and statistical analysis were clarified. A decision tree and checklist are provided to guide researchers through essential steps in conducting a cluster RCT. DISCUSSION: Cluster RCTs present special challenges in relation to design, conduct, and analysis. Nevertheless, they are an appropriate and potentially powerful tool for nursing research. With careful attention to the issues addressed in this article, researchers can use this approach successfully. PMID- 19289935 TI - Testing an alternate informed consent process. AB - BACKGROUND: One of the main problems in conducting clinical trials is low participation rate due to potential participants' misunderstanding of the rationale for the clinical trial or perceptions of loss of control over treatment decisions. OBJECTIVES: The objective of this study was to test an alternate informed consent process in cardiac rehabilitation participants that involved the use of a multimedia flip chart to describe a future randomized clinical trial and then asked, hypothetically, if they would participate in the future trial. METHODS: An attractive and inviting visual presentation of the study was created in the form of a 23-page flip chart that included 24 color photographs displaying information about the purpose of the study, similarities and differences between the two treatment groups, and the data collection process. We tested the flip chart in 35 cardiac rehabilitation participants. Participants were asked if they would participate in this future study on two occasions: immediately after the description of the flip chart and 24 hours later, after reading through the informed consent document. Participants were also asked their perceptions of the flip chart and consent process. RESULTS: Of the 35 participants surveyed, 19 (54%) indicated that they would participate in the future study. No participant changed his or her decision 24 hours later after reading the full consent form. The participation rate improved 145% over that of an earlier feasibility study where the recruitment rate was 22%. Most participants stated that the flip chart was helpful and informative and that the photographs were effective in communicating the purpose of the study. DISCUSSION: Participation rates could be enhanced in future clinical trials by using a visual presentation to explain and describe the study as part of the informed consent process. More research is needed to test alternate methods of obtaining informed consent. PMID- 19289936 TI - Effects of an analgesic trial in reducing pain behaviors in community-dwelling older adults with dementia. AB - BACKGROUND: Analgesic trials have been advocated as a means to reduce pain in persons with dementia (PWDs), but few systematic studies have been reported. OBJECTIVES: The aims of this study were to assess the feasibility of conducting a repeated-measures study of pain in PWDs and to investigate the effect of the scheduled dosing of acetaminophen in reducing observable pain behaviors in community-dwelling PWDs. METHODS: A within-subjects ABAB withdrawal design was utilized in this pilot study, with data collected daily for 24 days. Two baseline (nontreatment) phases were alternated with two treatment phases to examine the effects of scheduled extended-release acetaminophen (1.3 g every 8 hr) in reducing pain-related behaviors. The sample consisted of 3 participants (2 women; mean age = 85 years; mean Mini-Mental State Examination score = 11.7) with osteoarthritis. Pain behaviors were measured during a daily, videotaped activity based protocol designed to elicit pain. RESULTS: The results indicate that it is possible to conduct this type of intensive, daily pain investigation in community dwelling older adults with moderate to severe dementia. Behavioral indicators of pain varied daily and in response to treatment. During treatment phases, pain behaviors decreased in both frequency and duration relative to the control and baseline phases and increased when treatment was withdrawn. DISCUSSION: Acetaminophen use reduced pain behaviors associated with musculoskeletal pain in this sample of PWDs. This provides preliminary evidence that acetaminophen is an effective pain reliever for this population. In addition, the study supports the use of a within-subjects design to examine daily variability in pain behaviors and to demonstrate treatment effects in PWDs. PMID- 19289938 TI - Renewal ... inspiration is everywhere. PMID- 19289941 TI - Modern concepts of beauty. AB - Since the beginning of humanity, man has sought to define beauty. Whether by philosophy, mathematical constructs, social studies, or biology, the fascination with and the study of beauty has consumed our emotions and intellect. Many myths about this powerful force in our daily lives have been created, and some continue to be believed. Recent studies are shaping a new reality of beauty, one founded on evolutionary and biologic findings that link our fascination with beauty to our reproductive success. Cosmetic surgery, itself, may be an ongoing part of our biologic adaptation. PMID- 19289942 TI - Visual disfigurement and depression. AB - Approximately 10% of the population has a facial disfigurement, such as a scar, blemish, or deformity that severely affects the ability to lead a normal life, and 2 to 3% have a clearly visible blemish. They may experience depressive symptoms due to disfigurement, stressful life events, or other causes. Depression is a painful and costly disorder that is often unrecognized and untreated in specialty practices; it is linked with higher costs of care, lengths of stay, and rates of rehospitalization. Often, these individuals seek plastic surgery to repair the disfigurement, and depressive symptoms are not uncommon preoperatively, perioperatively, and postoperatively. In addition, depressive disorders exist among 20 to 32% of people with a medical disease. Major depression is a recurring and disabling illness that typically responds to treatment with psychotherapy, antidepressants, and social support. Nurses have a major role to play in screening for and detecting depression so it can be evaluated and referred for treatment. Nurses also provide education, psychosocial support, and advocacy for patients with depression. Identifying those with depressed symptoms allows the nurse to recommend treatment, offer referrals, and provide supportive interventions. PMID- 19289944 TI - A pediatric civil war burn victim: the patient's and caregivers' stress and coping. AB - A patient's and the caregivers' threats and reliefs during nursing of a pediatric civil war burn victim, treated through a charity grant in a Western hospital, are discussed and supported by a literature review. PMID- 19289945 TI - Psychological complications: national plastic surgical nursing survey. AB - An understanding of the scope and nature of negative psychological complications resulting from plastic surgery is vital. This knowledge is essential to improve the complex interactions that patients experience as part of their surgical intervention. This is a national survey of patients' psychological complications reported by 312 board certified plastic surgical nurses. Perceived patient psychological complications were reported at a higher rate than perceived physical complications (p> .001), with peri-operative complications of anxiety and mild depression being most prevalent. Less frequent psychological complications included patient disappointment and severe depression. Not surprisingly, surgical site pain and nonspecific physical distress were the most frequent physical complications; sleep disturbance was common. Psychological complications occur at higher rates than do physical complications in plastic surgical practices. Patients with pre-existing psychological conditions are more at risk for peri-operative psychological complications. Nurses should be adequately prepared to screen, support, and coordinate psychological treatment for plastic surgery patients with the plastic surgeon. PMID- 19289947 TI - Adjustment issues related to bilateral prophylactic mastectomy in women at elevated risk of developing breast cancer. AB - Breast cancer is the most common cancer diagnosis for North American and Western European women. Increased knowledge in availability of genetic testing has helped to identify those women at high risk of eventually developing breast cancer. This has resulted in more women considering bilateral prophylactic mastectomy as a viable preventative option. Although the efficacy of the procedure has been established, much less is known about the psychological impact of undergoing this procedure. In order to assess these factors, we reviewed what is currently known about the psychological impact of undergoing bilateral prophylactic mastectomy. Searches were conducted and inclusion criteria revealed articles that focused on the psychological components involved with undergoing a bilateral prophylactic mastectomy. The findings are summarized and fell broadly into one or more of the following three areas: (1) satisfaction or regret following the surgery, (2) psychosocial functioning after the surgery, and (3) predictors of quality of life. Plastic surgery nurses have a unique opportunity to impact important psychological considerations, such as expectations of the early postoperative period, body image concerns, and psychological distress. PMID- 19289948 TI - Women's perception of life following breast reduction: a phenomenological study. AB - PROBLEM: Little is known about the lived experiences of women following breast reduction surgery. Many studies addressed surgical technique or physical symptoms, but few looked at quality-of-life issues. AIMS: The aim of this qualitative study was to understand subjective issues of life after breast reduction surgery. Increased knowledge about this group of women may direct or guide nursing practice. PROCEDURE: A convenience sample of 9 English-speaking women aged 23-53 years were recruited during their follow-up visit. Patients were asked, "What is life like following your breast reduction?" ANALYSIS: A phenomenological approach was used to provide a rigorous, critical, and systematic study of the phenomenon. Analysis involved looking for themes. Giorgi's method guided the analysis. FINDINGS: Themes that emerged were improved physical health, increased self-esteem, self-confidence, and enhanced body image. In addition, preoperative expectations of the postoperative period varied from those of actual recovery, creating issues for some women. SIGNIFICANCE: Following breast reduction surgery, women experienced enhanced quality of life and improved physical symptoms. PMID- 19289950 TI - Psychological considerations in cosmetic breast augmentation. AB - Within the past decade, the popularity of cosmetic breast augmentation has surged and, with it, the interest in the psychological aspects of the procedure. Investigations of women who seek cosmetic breast augmentation have examined both their psychosocial characteristics and their motivations for surgery. Dissatisfaction both with body image and with breast size and/or shape are thought to be primary motivators for surgery. It is common for women seeking cosmetic breast augmentation to have some body image dissatisfaction. However, a considerable minority may suffer from excessive dissatisfaction consistent with the psychiatric diagnosis of body dysmorphic disorder, which is believed to contraindicate cosmetic surgery. Following breast augmentation, most women report satisfaction with the aesthetic result and improvements in body image. The impact of the procedure on other areas of functioning, such as self-esteem and quality of life, is less clear. These positive outcomes have been tempered by recent epidemiological studies that have identified a relationship between cosmetic breast implants and suicide. This article reviews this literature and provides recommendations to plastic surgical nurses regarding the psychological assessment and management of patients seeking breast augmentation. PMID- 19289953 TI - Preoperative assessment of eating disorders in plastic surgery patients. PMID- 19289954 TI - Compliance and the perioperative nurse. AB - While compliance efforts often occur "behind the scenes," there is much more to compliance than many people believe. This article will provide a brief glimpse at the components of a compliance program and some of the Office of Inspector General-targeted areas that may affect those in the perioperative area. PMID- 19289955 TI - The Wada test. AB - PURPOSE OF REVIEW: To examine the current clinical indications for conducting a Wada test in the presurgical evaluation of epilepsy surgery candidates in the light of research on the reliability and validity of proposed, noninvasive alternatives. RECENT FINDINGS: There has been a significant shift in the role of the Wada test in epilepsy surgery programmes. The majority of epilepsy centres no longer conduct a Wada test on every surgical candidate. The lateralization of language via invasive methods may not be necessary for temporal lobe epilepsy patients in whom a tailored resection will spare areas associated with language function. Functional MRI is being used in some centres to lateralize and localize language function in epilepsy surgery candidates. Magnetoencephalography also shows promise in this regard. Patients at high risk of a postoperative memory decline can be identified via multivariate models that utilize noninvasive measures of cerebral function and structure, together with demographic and clinical variables. SUMMARY: The clinical indications for a Wada test should be determined on a case-by-case basis, with careful consideration of the available noninvasive alternatives, to ensure that the risk-benefit ratio is appropriate for every patient. PMID- 19289956 TI - Bias from matching on age at death or censor in nested case-control studies. AB - BACKGROUND: Nested case-control studies frequently use incidence-density sampling based on attained age when matching controls to cases. A recently suggested additional matching criterion is age at death, with eligible controls having an age at death or censor within a specified number of years of the case's age at death. We simulated occupational cohorts with time-dependent exposures to evaluate whether adding this criterion introduces bias, and we investigated alternative methods of treating workers with zero exposure because of latency assumptions (ie, "lagged-out"). METHODS: We used simulated cohorts to consider null, positive, and negative exposure effects and lag periods of 0 and 10 years. Risk sets were constructed using incidence-density sampling with matching on attained age alone or attained age plus age at death. We estimated exposure effects using conditional logistic regression for unlagged and 10-year lagged cumulative exposure. Lagged-out workers were either excluded or included and assigned zero exposure. RESULTS: Effect estimates were generally unbiased when controls were selected by matching on attained age alone. However, the estimates were downwardly biased under the additional matching criterion. When risk was related to a lagged cumulative exposure, estimates including lagged-out workers were similarly or less biased than those excluding lagged-out workers. CONCLUSIONS: In these simulations, incidence-density sampling with matching on attained age plus age at death introduced bias. This is because sampled controls were younger at first exposure, with higher cumulative exposure compared with controls selected by matching on attained age alone. Incidence-density sampling with matching on attained age alone (and including lagged-out workers) did not introduce bias. PMID- 19289957 TI - High levels of depressive symptoms in pregnancy with low omega-3 fatty acid intake from fish. AB - BACKGROUND: Depression during pregnancy has adverse consequences for both mother and child. Although common in western countries, depression appears to be virtually absent in countries with high seafood intake. We test the hypothesis that low seafood intake during pregnancy is associated with increased prevalence of depressive symptoms. METHODS: This study used data prospectively collected from women participating in the Avon Longitudinal Study of Parents and Children in the period 1991-1992. At 32 weeks' gestation, the mother completed a questionnaire that included symptoms of depression and a food frequency questionnaire from which the amount of omega-3 fatty acids from fish was calculated. Statistical analysis took social and lifestyle factors into account. RESULTS: Unadjusted and adjusted analyses showed lower maternal intake of omega-3 from seafood was associated with high levels of depressive symptoms. Compared with women consuming more than 1.5 g omega-3 from seafood per week, those consuming none were more likely to have high levels of depressive symptoms at 32 weeks' gestation (adjusted odds ratios = 1.54; 95% confidence interval = 1.25 1.89). CONCLUSIONS: These observational data support an association between low omega-3 intake from seafood and increased risk of high levels of depressive symptoms during pregnancy. Eating seafood during pregnancy may have beneficial effects on mental well-being. PMID- 19289958 TI - Unexpected lower prevalence of depression in patients with diabetes: selection bias in a waiting room population. AB - BACKGROUND: We initially studied the association between diabetes and depression in general practice attendees. Subsequently, we investigated whether our findings were influenced by selection bias. METHODS: Unexpectedly, the prevalence of depression was lower in diabetes patients (4%) than in subjects without diabetes (14%). To explore the possibility of selection bias, we first described the selection process from the general population to our study population and provided prevalence data of diabetes and depression in the different populations based on literature and our data. Second, we performed a sensitivity analysis and described possible reasons for selection. Third, we studied the association between other chronic diseases and depression. RESULTS: These analyses suggested that the lower prevalence of depression in diabetes patients was due to selection bias. Visiting the general practitioner for a control visit seemed to play a role in this selection process. CONCLUSIONS: This study illustrated the potential for selection bias in a waiting room population. The degree of bias depended on the exposure under study. PMID- 19289959 TI - Logistic regression with incomplete covariate data in complex survey sampling: application of reweighted estimating equations. AB - Weighted survey data with missing data for some covariates presents a substantial challenge for analysis. We addressed this problem by using a reweighting technique in a logistic regression model to estimate parameters. Each survey weight was adjusted by the inverse of the probability that the possibly missing covariate was observed. The reweighted estimating equations procedure was compared with a complete case analysis (after discarding any subjects with missing data) in a simulation study to assess bias reduction. The method was also applied to data obtained from a national health survey (National Health and Nutritional Examination Survey or NHANES). Adjusting the sampling weights by the inverse probability of being completely observed appears to be effective in accounting for missing data and reducing the bias of the complete case estimate of the regression coefficients. PMID- 19289960 TI - Waist-height ratio as a predictor of coronary heart disease among women. AB - BACKGROUND: It is not yet established what specific measures of obesity might be most strongly associated with risk of coronary artery disease. We compared the waist-height ratio to waist-hip ratio, waist circumference, and body mass index as predictors of subsequent coronary heart disease (CHD) in a group of predominantly postmenopausal women. METHODS: The data come from the prospective Nurses' Health Study cohort. We included 45,563 women in 1986 who were aged 40-65 years and were free of heart disease, stroke, and cancer. Waist circumference, hip circumference, height, weight, age, and other covariates were collected by questionnaire. Our primary end point was incident coronary heart disease reported up to June 2002. Areas under the receiver operating characteristic curves (AUCs) were estimated nonparametrically for each of the anthropometric measures. We estimated differences between the AUCs for weight-height ratio and the other measures, with corresponding 95% confidence intervals. We used Cox proportional hazard models to estimate the relationships with risk of CHD. RESULTS: Waist height ratio, waist-hip ratio, and waist circumference were similar in predicting subsequent risk of CHD. All 3 waist-derived measures were superior to body-mass index (BMI) in predicting CHD. The unadjusted AUCs were 0.62 (95% confidence interval = 0.60-0.64) for waist-height ratio, 0.63 (0.61-0.65) for waist-hip ratio, 0.62 (0.60-0.64) for waist-circumference, and 0.57 (0.55-0.59) for BMI. CONCLUSION: Waist-height ratio is comparable with waist circumference and waist hip ratio for prediction of coronary heart disease incidence among middle-aged and older women, but superior to BMI. PMID- 19289961 TI - Bias in full cohort and nested case-control studies? PMID- 19289962 TI - Health effects of long-term air pollution: influence of exposure prediction methods. AB - BACKGROUND: Air pollution studies increasingly estimate individual-level exposures from area-based measurements by using exposure prediction methods such as nearest-monitor and kriging predictions. However, little is known about the properties of these methods for health effects estimation. This simulation study explores how 2 common prediction approaches for fine particulate matter (PM2.5) affect relative risk estimates for cardiovascular events in a single geographic area. METHODS: We estimated 2 sets of parameters to define correlation structures from 2002 data on PM2.5 in the Los Angeles area, and selected additional parameters to evaluate various correlation features. For each structure, annual average PM2.5 was generated at 22 monitoring sites and 2000 preselected individual locations in Los Angeles. Associated survival time until cardiovascular event was simulated for 10,000 hypothetical subjects. Using PM2.5 generated at monitoring sites, we predicted PM2.5 at subject locations by nearest monitor and kriging interpolation. Finally, we estimated relative risks of the effect of PM2.5 on time to cardiovascular event. RESULTS: Health effect estimates for cardiovascular events had higher or similar coverage probability for kriging compared with nearest-monitor exposures. The lower mean square error of nearest monitor prediction resulted from more precise but biased health effect estimates. The difference between these approaches dramatically moderated when spatial correlation increased and geographic characteristics were included in the mean model. CONCLUSIONS: When the underlying exposure distribution has a large amount of spatial dependence, both kriging and nearest-monitor predictions gave good health effect estimates. For exposure with little spatial dependence, kriging exposure was preferable but gave very uncertain estimates. PMID- 19289963 TI - Are nested case-control studies biased? AB - It has been recently asserted that the nested case-control study design, in which case-control sets are sampled from cohort risk sets, can introduce bias ("study design bias") when there are lagged exposures. The bases for this claim include a theoretical and an "empirical evaluation" argument. We examined both of these arguments and found them to be incorrect. We describe an appropriate empirical evaluation method to explore the performance of nested case-control study designs and analysis methods from an existing cohort. This empirical evaluation approach relies on simulating case-control outcomes from risk sets in the cohort from which the case-control study is to be performed. Because it is based on the underlying cohort structure, the empirical evaluation can provide an assessment that is tailored to the specific characteristics of the study under consideration. The methods are illustrated using samples from the Colorado Plateau uranium miners cohort. PMID- 19289964 TI - Maternal smoking and infant mortality: does quitting smoking reduce the risk of infant death? AB - BACKGROUND: Maternal smoking has repeatedly been associated with increased infant mortality rates. No study has investigated whether smoking cessation influences the risk of infant death. This study estimates infant mortality after the second pregnancy in relation to smoking behavior in both the first and the second pregnancy. METHODS: We used the Swedish Medical Birth Register to identify women who delivered their first and second singleton infants during 1983-2002. Maternal smoking during the 2 pregnancies was categorized into (1) never smoker, (2) quitter, (3) starter, and (4) persistent smoker. In the second pregnancy, 555,046 live births (of at least 22 completed gestational weeks) were followed for infant death within 1 year. Cox regression was used to estimate hazard ratios (HRs) with 95% confidence intervals (CIs). RESULTS: Compared with infants born to never smokers, the HR (95% CI) of infant mortality in the second pregnancy was 2.0 (1.7 2.4) among infants born to persistently heavy smokers, whereas among women who stopped smoking in the second pregnancy, the HRs were 1.4 (1.0-2.0) among those who had been heavy smokers in the first pregnancy, and 1.0 (0.8-1.2) among those who had been light smokers. The association of smoking during pregnancy with infant mortality was modified by infant's age, and was strongest at 4-15 weeks after birth. The smoking effect on neonatal mortality, but not postneonatal mortality, was mediated by gestational age. CONCLUSIONS: Smoking cessation reduced the risk of infant death. The smoking-related risk of neonatal mortality appears to be mediated by smoking effects on gestational age, a factor that only partly explains the association between smoking and postneonatal mortality. PMID- 19289965 TI - Elemental carbon exposure at residence and survival after acute myocardial infarction. AB - BACKGROUND: Particulate air pollution has been consistently related to cardiovascular mortality. Some evidence suggests that particulate matter may accelerate the atherosclerotic process. Effects of within-city variations of particulate air pollution on survival after an acute cardiovascular event have been little explored. METHODS: We conducted a cohort study of hospital survivors of acute myocardial infarction (MI) from the Worcester, MA, metropolitan area to investigate the long-term effects of within-city variation in traffic-related air pollution on mortality. The study builds on an ongoing community-wide investigation examining changes over time in MI incidence and case-fatality rates. We included confirmed cases of MI in 1995, 1997, 1999, 2001, and 2003. Long-term survival status was ascertained through 2005. A validated spatiotemporal land use regression model for traffic-related air pollution was developed and annual averages of elemental carbon at residence estimated. The effect of estimated elemental carbon on the long-term mortality of patients discharged after MI was analyzed using a Cox proportional hazards model, controlling for a variety of demographic, medical history, and clinical variables. RESULTS: Of the 3895 patients with validated MI, 44% died during follow-up. Exposure to estimated elemental carbon in the year of entry into the study was 0.44 microg/m on average. All-cause mortality increased by 15% (95% confidence interval = 0.03%-29%) per interquartile range increase in estimated yearly elemental carbon (0.24 microg/m) after the second year of survival. No association between traffic-related pollution and all-cause mortality was observed during the first 2 years of follow-up. CONCLUSIONS: Chronic traffic related particulate air pollution is associated with increased mortality in hospital survivors of acute MI after the second year of survival. PMID- 19289966 TI - Maternal blood manganese levels and infant birth weight. AB - BACKGROUND: Manganese is both an essential element and a known neurotoxicant to children. High manganese exposures have been associated with negative reproductive outcomes in animals, but few epidemiologic studies have examined the effects of human fetal manganese exposure. METHODS: We studied the association between maternal and umbilical cord blood manganese levels and birth weight in a cohort of 470 mother-infant pairs born at term (>or=37 weeks gestation) in Ottawa County, Oklahoma. Nonlinear spline and quadratic regression models were used to test the hypothesis of an inverted U-shaped relationship between manganese levels and birth weight. RESULTS: Mean (standard deviation) concentration of manganese was 2.4 (0.95) microg/dL in the maternal blood and 4.2 (1.6) microg/dL in the cord blood. Umbilical cord manganese was not associated with birth weight. A nonlinear relationship was observed between maternal manganese and birth weight after adjusting for potential confounders. Birth weight increased with manganese levels up to 3.1 microg/L, and then a slight reduction in weight was observed at higher levels. Compared with the 3.1-microg/L point of inflection, birth weight estimates at the 5th (1.3 microg/L) and 95th (4.0 microg/L) percentiles of exposure were -160 g (95% confidence interval = -286 to -33) and -46 g (-38 to 131), respectively. CONCLUSIONS: Maternal blood manganese levels during pregnancy are associated with birth weight in a nonlinear pattern in full-term infants. These findings suggest that manganese may affect fetal growth. Possible detrimental effects of elevated manganese levels on the fetus should be further examined in more highly exposed populations. PMID- 19289967 TI - Uncontrolled migrations as a cause of inequality in health and mortality in small area studies. AB - Mortality is one of the most widely used indicators in small-area ecologic studies. Both accessibility to mortality data and advances in the development of new disease mapping techniques have contributed to an abundance of mortality maps and atlases over the last decade. Results may be biased in this kind of study if there has been unmeasured geographic mobility of the population. Most published papers tend to neglect this possibility. We use the theory of dynamics systems to demonstrate that migratory flows unmonitored by official population registers may lead to major errors in mortality rates and relative risks. Simulations in 4 scenarios showed more than 8% underestimation of the mortality rate and more than 11% underestimation of relative risk in areas with high uncontrolled emigration, and above 19% overestimation of mortality rate and above 15% overestimation of relative risk in areas with high uncontrolled immigration.Researchers conducting small-area epidemiologic studies should explore the reliability of population information in geographic areas before drawing hypothesis or conclusions on other possible causes of mortality differences. PMID- 19289969 TI - Factors associated with an explosive HIV epidemic among injecting drug users in Sargodha, Pakistan. AB - BACKGROUND: During a routine surveillance round, an extremely high prevalence of 51.3% was found among injecting drug users (IDUs) in Sargodha, a small town in Pakistan. This sharp increase of HIV among this group warranted an urgent need to explore the factors leading toward this explosion of HIV infection among IDUs at this very location to direct the policy makers in designing preventive activities, especially in the context of HIV prevention. METHODS: In 2007, 400 current IDUs were recruited through multistage cluster sampling based on mapping studies. Participants provided dried blood spot samples for HIV testing and completed a questionnaire on demographic characteristics and sexual/drug injecting behaviors. Logistic regression was used to examine the independent association of multiple variables with HIV infection. RESULTS: Of the 400 IDUs tested, 205 (51.3%) were HIV positive confirmed through enzyme-linked immunosorbent assay and Western blot. In multivariate analysis, geographical location of IDUs [adjusted odds ratio (aOR) 2.4 for IDUs located in the northern zone vs southern zone, 95% confidence interval (CI) 1.5 to 3.7], injected in groups (aOR 1.8, 95% CI 1.1 to 3.1), and sharing injecting paraphernalia with other IDUs (aOR 1.5, 95% CI 1.0 to 2.4) were strong correlates of HIV infection. CONCLUSIONS: Effective outreach programs need to be developed to provide a comprehensive package of HIV prevention services to IDUs not reached by existing services, entry into drug abuse treatment and medical care needs to be facilitated, and essential legal and social services need to be provided with community participation. PMID- 19289970 TI - Inhibition of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase reduces leukocyte recruitment and hepatocyte apoptosis in endotoxin-induced liver injury. AB - BACKGROUND: Endotoxemia is well known to be associated with an excessive host response to bacteria or microbial compounds, resulting in systemic inflammation and organ injury. The aim of the present study was to examine the effects of simvastatin on endotoxemic liver injury. METHODS: Male C57BL/6J mice were challenged intraperitoneally with 0.5 mg/kg Escherichia coli-lipopolysaccharide (LPS) and 0.9 g/kg d-galactosamine (Gal). Mice were pretreated with 0.2 mg/kg simvastatin. Lipopolysaccharide/d-Gal-injected mice without simvastatin served as endotoxemic controls, and sham mice served as negative controls. Additional mice were challenged with LPS/d-Gal and co-treated with simvastatin and 10 mg/kg mevalonate to determine the role of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase. After 6 hours of endotoxemia serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities as well as caspase-3 and myeloperoxidase activity were determined. RESULTS: Endotoxemia caused a substantial hepatocellular injury as indicated by significantly elevated serum ALT and AST levels and hepatocellular apoptosis. Leukocyte infiltration in the liver was significantly elevated in endotoxemic mice. Simvastatin significantly reduced endotoxin-induced hepatocellular damage and apoptosis. Moreover, hepatic accumulation of leukocytes was attenuated by simvastatin in endotoxemic animals. Co-administration of mevalonate abolished protective effects of simvastatin on endotoxin-provoked increases in ALT, AST, and hepatocellular apoptosis as well as leukocyte recruitment. CONCLUSIONS: Simvastatin has the capacity to prevent endotoxemic liver injury by inhibiting leukocyte infiltration and hepatocellular apoptosis. These protective effects exerted by simvastatin are dependent on the 3 hydroxy-3-methyl-glutaryl-coenzyme A reductase pathway. Thus, simvastatin may represent a potential approach to prevent endotoxemia-associated liver dysfunction. PMID- 19289971 TI - The academic paradigm is the problem. PMID- 19289972 TI - Disruption of cholesterol efflux by coxib medications and inflammatory processes: link to increased cardiovascular risk. AB - Atherosclerosis is a chronic progressive disease that is a major contributor to cardiac death. It is characterized by inflammation and cholesterol deposition in the arterial wall. Excess cholesterol accumulation occurs as a result of an imbalance between delivery and removal and leads to formation of lipid-laden foam cells. Removal of cholesterol through a process known as reverse cholesterol transport requires the coordinated functioning of a number of genes including the P450 27-hydroxylase and the adenosine triphosphate-binding cassette transporter A1 (ABCA1). Reverse cholesterol transport is a key defense against atheroma formation. This review discusses the role of inflammatory processes in impeding reverse cholesterol transport. Particular emphasis is placed on the disruption of cholesterol outflow observed in the presence of cyclooxygenase inhibitors in cultured monocytes/macrophages. These inhibitors, which are used clinically to relieve pain and inflammation, have been associated with increased risk of cardiovascular disease and myocardial infarction. We explore the relationship between suppression of reverse cholesterol transport and harmful cardiac effects of coxibs. PMID- 19289975 TI - Refractive errors and amblyopia in children entering school: Shahrood, Iran. AB - PURPOSE: To determine the prevalence of amblyopia, strabismus, and refractive errors in children entering school. METHODS: In this cross-sectional population based study, 6-year-old children enrolling in Shahrood schools were randomly sampled. Those with organic blindness or mental retardation were excluded. Ocular alignment, visual acuity, stereopsis, cover testing, and cycloplegic refraction were recorded for all children by an optometrist. A diagnosis of amblyopia was based on a best-corrected visual acuity of 6/12 or less in one or both eyes, or a bilateral difference of at least two best-corrected visual acuity lines. RESULTS: A total of 902 children were invited for optometry examinations, among which 827 (91.7%) responded and 815 children (98.5%) were included in the study. The prevalence of significantly reduced visual acuity (uncorrected VA < or = 6/12 in either eye) and amblyopia was 3.6 and 1.7%, respectively. The prevalence of hyperopia (+2.00 D or worse), myopia (-0.50 D or worse), astigmatism (0.75 D or worse), and anisometropia (1.00 D or more) were 20.5, 1.7, 19.6, and 2.2%, respectively. Significant refractive error, defined as hyperopia >+3.50 D, myopia >-3.00 D, astigmatism >1.50 D in the orthogonal meridian or >1.00 D in the oblique meridian, or anisometropia (hyperopic >1.00 D, myopic >3.00 D) was detected in 2.1, 0.1, 5.0, and 0.9% of the sample, respectively. Strabismus was diagnosed in 1.2% of children. Overall, 52 children (6.4%; 95% confidence interval [CI], 4.7-8.1%) were at risk of amblyopia; of these, 81% had significant refractive errors, 11% had strabismus, and 8% had both. CONCLUSION: A considerable proportion of 6-year-old children have strabismus and/or significant, potentially amblyogenic refractive errors. The relatively high rate of hyperopia and astigmatism in the studied population needs more attention. The results of the study emphasize the need for adequate diagnostic and therapeutic eye care services for preschool children. PMID- 19289976 TI - Changes in UV-visible transmittance of silicone-hydrogel contact lenses induced by wear. AB - PURPOSE: To evaluate the influence of wear of silicone-hydrogel contact lenses on lens ultraviolet and visible spectrum transmittance by studying several contact lenses before and after wear. METHOD: To investigate the ability of contact lenses to maintain their transmittance characteristics, we measured 104 different contact lenses in the ultraviolet (UV) and visible range from 200 to 700 nm, with a Shimadzu UV3101-PC UV-vis-NIR spectrophotometer equipped with an integrating sphere. The lenses used in this study were Acuvue Advance (Johnson & Johnson Vision Care, Inc.), Air Optix Night & Day (CIBA Vision), Air Optix (CIBA Vision), and PureVision (Bausch & Lomb). A conventional hydrogel contact lens was also tested, Acuvue (Johnson & Johnson Vision Care Inc.). RESULTS: Our study indicates that lenses that do not have UV absorbers incorporated into the polymer transmitted most of the UV radiation (UVR) before and after wear. The results of the statistical analysis show that for the UVC portion of the spectrum significant difference exists within the measurements obtained before and after wear for all the lenses, with the exception of PureVision. Acuvue Advance is the only material in which significant transmittance differences were observed in the visible spectral range. CONCLUSIONS: Transmittance is modified after contact lenses wear, probably due to the formation of biofilms on the contact lens surface, being more noticeable in the UVR region of the spectrum (200-400 nm). Silicone-hydrogel and conventional hydrogel contact lens materials that provide UVR protection (UV-blocker) maintain this property even after being worn. The changes observed in the visible spectrum seem not to have any implications in visual performance of silicone-hydrogel contact lenses. PMID- 19289978 TI - Live attenuated human rotavirus vaccine, RIX4414, provides clinical protection in infants against rotavirus strains with and without shared G and P genotypes: integrated analysis of randomized controlled trials. AB - BACKGROUND: : The 2-dose, oral live attenuated human G1P[8] rotavirus vaccine (RIX4414) is highly effective against rotavirus gastroenteritis caused by circulating G1 and non-G1 types. An integrated analysis on vaccine efficacy was undertaken to obtain more precise estimates of the overall protective effect of the RIX4414 vaccine against rotavirus gastroenteritis due to common rotavirus types (G1, G3, G4, G9, P[8]) and less commonly encountered strains such as G2P[4] across heterogenous settings. METHODS: : The studies used in the integrated analysis were all previously reported randomized, double-blind, placebo controlled, phase II and III trials with at least 1 report of rotavirus gastroenteritis in the efficacy follow-up period (up to 1 year of age or end of first RV epidemic season after vaccination). The integrated analysis was performed for all circulating rotavirus strains sharing G and/or P genotype and not sharing G or P genotype with the vaccine strain. Vaccine efficacy was estimated as 1 minus rate of rotavirus gastroenteritis relative to placebo, using exact Poisson rate ratio stratified by study. RESULTS: : The integrated estimates for vaccine efficacy against severe rotavirus gastroenteritis were 87.43% (95% confidence interval [CI]: 78.89-92.86) for G1P[8] strains, 71.42% (95% CI: 20.12 91.11) for G2P[4] strains, 90.19% (95% CI: 55.51-98.94) for G3P[8] strains, 93.37% (95% CI: 51.50-99.85) for G4P[8] strains, and 83.76% (95% CI: 71.18-91.28) for G9P[8] strains. The integrated estimates for vaccine efficacies against rotavirus gastroenteritis of any severity were 82.57% (95% CI: 73.91-88.56) for G1P[8] strains, 81.04% (95% CI: 31.58-95.76) for G2P[4] strains, 87.66% (95% CI: 34.57-98.76) for G3P[8] strains, 84.86% (95% CI: 50.92-96.41) for G4P[8] strains, and 60.64% (95% CI: 38.15-74.96) for G9P[8] strains. CONCLUSIONS: : Two doses of RIX4414 provide overall good clinical protection against all cases of rotavirus gastroenteritis and comparable, high clinical protection against severe rotavirus gastroenteritis caused by circulating rotavirus strains with and without G and P genotypes shared with the vaccine strain, such as G2P[4]. PMID- 19289977 TI - Validity of the convergence insufficiency symptom survey: a confirmatory study. AB - PURPOSE: The objectives of the present study were to evaluate whether investigator bias influenced the Convergence Insufficiency Symptom Survey (CISS) scores of children with normal binocular vision (NBV) in our original validation study, reevaluate the usefulness of the cutoff score of 16, and reexamine the validity of the CISS. METHODS: Six clinical sites participating in the Convergence Insufficiency Treatment Trial (CITT) enrolled 46 children 9 to <18 years with NBV. Examiners masked to the child's binocular vision status administered the CISS. The mean CISS score was compared with that from the children with NBV in the original, unmasked CISS study and also to that of the 221 symptomatic convergence insufficiency (CI) children enrolled in the CITT. RESULTS: The mean (+/-standard deviation) CISS score for 46 subjects with NBV was 10.4 (+/-8.1). This was comparable with our prior unmasked NBV study (mean = 8.1 (+/-6.2); p = 0.11) but was significantly different from that of the CITT CI group (mean = 29.8 +/- 9.0; p < 0.001). Eighty-three percent of these NBV subjects scored <16 on the CISS, which is not statistically different from the 87.5% found in the original unmasked study (p = 0.49). CONCLUSIONS: Examiner bias did not affect the CISS scores for subjects with NBV in our prior study. The CISS continues to be a valid instrument for quantifying symptoms in 9 to <18-year-old children. These results also confirm the validity of a cut-point of > or = 16 in distinguishing children with symptomatic CI from those with NBV. PMID- 19289979 TI - Effectiveness of home-based management of newborn infections by community health workers in rural Bangladesh. AB - BACKGROUND: : Infections account for about half of neonatal deaths in low resource settings. Limited evidence supports home-based treatment of newborn infections by community health workers (CHW). METHODS: : In one study arm of a cluster randomized controlled trial, CHWs assessed neonates at home, using a 20 sign clinical algorithm and classified sick neonates as having very severe disease or possible very severe disease. Over a 2-year period, 10,585 live births were recorded in the study area. CHWs assessed 8474 (80%) of the neonates within the first week of life and referred neonates with signs of severe disease. If referral failed but parents consented to home treatment, CHWs treated neonates with very severe disease or possible very severe disease with multiple signs, using injectable antibiotics. RESULTS: : For very severe disease, referral compliance was 34% (162/478 cases), and home treatment acceptance was 43% (204/478 cases). The case fatality rate was 4.4% (9/204) for CHW treatment, 14.2% (23/162) for treatment by qualified medical providers, and 28.5% (32/112) for those who received no treatment or who were treated by other unqualified providers. After controlling for differences in background characteristics and illness signs among treatment groups, newborns treated by CHWs had a hazard ratio of 0.22 (95% confidence interval [CI] = 0.07-0.71) for death during the neonatal period and those treated by qualified providers had a hazard ratio of 0.61 (95% CI = 0.37-0.99), compared with newborns who received no treatment or were treated by untrained providers. Significantly increased hazards ratios of death were observed for neonates with convulsions (hazard ratio [HR] = 6.54; 95% CI = 3.98 10.76), chest in-drawing (HR = 2.38, 95% CI = 1.29-4.39), temperature <35.3 degrees C (HR = 3.47, 95% CI = 1.30-9.24), and unconsciousness (HR = 7.92, 95% CI = 3.13-20.04). CONCLUSIONS: : Home treatment of very severe disease in neonates by CHWs was effective and acceptable in a low-resource setting in Bangladesh. PMID- 19289980 TI - Clinical courses of children with acute lobar nephronia correlated with computed tomographic patterns. AB - BACKGROUND: : Acute lobar nephronia (ALN) is a complicated acute renal infection that is part of the progression of the inflammatory process of acute pyelonephritis that can progress to renal abscess if left untreated. We examined the place of ALN in the spectrum of upper urinary tract infections. METHODS: : The medical records of all patients diagnosed with ALN by computed tomography (CT) were reviewed retrospectively in terms of their demographic characteristics, clinical presentations, and laboratory findings. The patterns of the ALN lesions were checked on all contrast-enhanced CT films and the volume of the CT lesions (the extent of renal involvement) was estimated in every patient. Then, the correlation between the clinical presentation and CT lesions was examined. RESULTS: : Two distinct CT patterns of ALN lesions were observed in 127 patients: 94 simple ALN and 33 complicated ALN. Simple ALN appeared as striated or wedge shaped, poorly defined regions of homogeneously decreased nephrographic density, whereas the lesions of complicated ALN showed heterogeneously decreased nephrographic density after contrast enhancement. The volume fraction of the CT lesions correlated well with the duration of fever before (P = 0.006) and after (P < 0.001) treatment in patients with simple ALN, while only the correlation between the fever duration after treatment and the volume fraction of the CT lesions (P < 0.001) was significant in patients with complicated ALN. Patients with complicated ALN were generally older (P = 0.004) and febrile longer after starting antibiotic treatment (P < 0.001). All treatment failures were in the complicated ALN group. CONCLUSIONS: : Children with ALN comprise 2 groups with different clinical presentations and treatment responses: simple ALN and complicated ALN. We suggest that simple ALN be regarded as a continuation of acute pyelonephritis and require 2 weeks of antibiotic treatment, while complicated ALN is a distinct more severe form requiring a 3-week or longer therapy regimen as the treatment of choice. PMID- 19289981 TI - Failure of human immunodeficiency virus enzyme immunoassay to rule out infection among polymerase chain reaction-negative Vietnamese infants at 12 months of age. AB - BACKGROUND: : Previous studies have demonstrated that >90% of HIV-uninfected infants serorevert, as seen in the results of enzyme immunoassay (EIA) testing by 12 months of age, making it feasible to confirm or rule out infection. We assessed the reliability of EIA in a cohort of Vietnamese infants. METHODS: : HIV exposed, uninfected infants enrolled in a parent diagnostic and monitoring study from February 2005 through August 2006 were eligible for inclusion in a prospective cohort study of HIV-EIA performance. Testing using 2 standard assays (Genscreen HIV 1/2 version 2, Bio-Rad; Murex 1.2.0, Murex Biotech) was initiated at 12 months of age. Infants were categorized as EIA-negative (seroreverted; negative Genscreen), EIA-indeterminate (positive Genscreen, negative Murex), or EIA-positive (Genscreen and Murex positive). RESULTS: : Of 273 infants included in the study, 59 (22%) were EIA-negative at 12 months, 131 (48%) were indeterminate, and 83 (30%) were EIA-positive; specificity 21.6 (95% confidence interval: 16.6, 26.3). Infants with positive EIAs at 12 months were 74% more likely than EIA-indeterminate infants to test indeterminate or positive at 18 months (risk ratio, 1.74, 95% confidence interval: 1.15, 2.64; P = 0.03). CONCLUSIONS: : Expectations regarding infant seroreversion by standard EIAs should be reassessed to reflect potential cross-regional differences in their performance. PMID- 19289982 TI - Retinal sensitivity after intravitreal injection of bevacizumab for the treatment of macular edema secondary to retinal vein occlusion. AB - PURPOSE: To evaluate the change in macular function after intravitreal injection of bevacizumab for the treatment of macular edema associated with retinal vein occlusion. METHODS: For this interventional case series, 20 eyes of 20 patients with macular edema associated with retinal vein occlusion were treated with an intravitreal injection of bevacizumab. Microperimetry in the macular area was performed with a Micro Perimeter 1 before and at 1, 3, and 6 months after treatment. RESULTS: Improvement in macular function was detected immediately after treatment and lasted for at least 6 months. As measured by the Micro Perimeter 1, mean retinal sensitivities within the central 10 degrees field (4.9 +/- 2.7 dB at baseline) improved to 7.2 +/- 3.1 dB at 1 month, to 7.6 +/- 3.4 dB at 3 months, and to 7.7 +/- 3.9 dB at 6 months (P < 0.001). Of the 20 eyes, a recurrence of macular edema was observed in 14 (70%), but with the use of optical coherence tomography, integrity of the outer aspect of the foveal photoreceptors was detected at 3 months to 6 months in 15 (75%) eyes. CONCLUSION: In eyes with macular edema associated with retinal vein occlusion, improvement in macular function was detected immediately after intravitreal injection of bevacizumab and lasted for at least 6 months. PMID- 19289983 TI - Funduscopic and angiographic appearance in the neuronal ceroid lipofuscinoses. AB - PURPOSE: To characterize the retinal features of neuronal ceroid lipofuscinoses (NCLs) and to determine if retinal abnormalities are detectable in carriers of these autosomal recessively inherited diseases. METHODS: Carriers of the NCLs and their affected children underwent ophthalmic examination including color fundus photography in all patients and fluorescein angiography in selected patients. Twenty-nine patients with NCL were examined and photographed: 3 with infantile form, 2 with late-infantile form, and 24 with juvenile form. Fourteen patients underwent fluorescein angiography. RESULTS: Infantile and late-infantile retinal findings include fine retinal pigment epithelium pigment atrophy with no bone spicule changes and disk pallor. Juvenile retinal findings include macular retinal pigment epithelium atrophy and pigment stippling (>50%), epiretinal membrane (33%), bull's eye maculopathy (25%), and peripheral bone spicules (46%) and variable disk pallor. Fluorescein angiography of juvenile patients demonstrated diffuse retinal pigment epithelium atrophy with stippled hyperfluorescence (93%). Heterozygous NCL carriers had no identifying retinal abnormalities. CONCLUSION: Significant variability exists in the retinal appearance of the NCLs, but, in general, ophthalmoscopy and fluorescein angiography distinguish these patients from other more common blinding disorders of childhood such as retinitis pigmentosa and Stargardt disease. Examining retinas of parents of affected children does not aid in the diagnosis of NCL. PMID- 19289984 TI - Effect of ranibizumab retreatment frequency on neurosensory retinal volume in neovascular amd. AB - PURPOSE: To determine the characteristics of patients with neovascular age related macular degeneration who show initial anatomic improvements on optical coherence tomography in response to treatment with ranibizumab, but who subsequently regress toward their anatomic baseline. METHODS: Data from 50 consecutive patients, receiving ranibizumab therapy for neovascular age-related macular degeneration, were collected. Raw StratusOCT images were analyzed using custom software ("OCTOR"). Changes in volume of neurosensory retina at months 1, 3, and 6 were calculated. Baseline demographic and morphologic characteristics were compared. RESULTS: Forty-two patients (84%) showed a reduction in total retinal volume 1 month after initial treatment with ranibizumab. Of the patients that initially showed a reduction, 16 (38%) maintained this reduction through month 6, whereas 26 patients (62%) demonstrated a subsequent increase in retinal volume. Patients who maintained a reduction in edema received 3.75 +/- 1.18 injections of ranibizumab versus 2.96 +/- 1.34 injections for patients who did not (P = 0.049). Regression of initial anatomic improvements was associated with worsening of visual acuity (r = 0.599, P = 0.002). CONCLUSION: Patients receiving fewer injections of ranibizumab appeared less likely to maintain anatomic improvements achieved following commencement of ranibizumab therapy; regression of these improvements was associated with deterioration in visual acuity. PMID- 19289985 TI - Peripheral 360 degrees retinectomy in complex retinal detachment. AB - PURPOSE: To report the functional and anatomical results and complications of 360 degrees peripheral retinectomy for management of complicated retinal detachment. METHODS: Patients with complicated retinal detachment underwent pars plana vitrectomy, 360 degrees retinectomy, intraoperative endolaser, and internal tamponade with silicone oil. Postoperative visual acuity, intraocular pressure, retinal status, need for reoperation, and complications are presented. RESULTS: Twenty eyes of 19 patients with a mean age of 32.4 years (8-75 years) underwent pars plana vitrectomy and 360 degrees peripheral retinectomy for complicated retinal detachment due to anterior proliferative vitreoretinopathy, unstable edge of retinal break, anterior hyaloidal fibrovascular proliferation, retinal incarceration in scleral wound, and 300 degrees giant retinal tear. Intraoperative reattachment was achieved in 18 eyes. Mean postoperative follow-up time was 24.2 months (2-70 months). Retina was attached in 14 eyes (70%) in the last visit. Eight eyes (40%) had 5/200 or greater visual acuity. Preoperative and postoperative visual acuities did not have significant correlation (Spearman correlation coefficient = 0.291). There was no relation between diagnosis and anatomical outcome (P > 0.2). CONCLUSION: Relaxing peripheral 360 degrees retinectomy is an effective procedure for flattening the retina in complicated retinal detachments when no other option is available. PMID- 19289986 TI - 25-gauge sutureless vitrectomy: variations in incision architecture. AB - BACKGROUND/PURPOSE: To examine the histologic architecture of 25-gauge transconjunctival sutureless vitrectomy incisions. METHODS: Four groups of sutureless incisions were constructed in cadaver eyes using a 25-gauge trocar cannula system. In Group 1, oblique incisions were constructed by inserting the trocar to the bevel and then turning vertically to enter the vitreous cavity. Oblique incisions in Group 2 were constructed as in Group 1, but the trocar was inserted to the beginning of the 25-gauge cannula. Incisions in Group 3 were constructed as in Group 2, but before removing the cannula from the eye vitrectomy was performed. Incisions in Group 4 were constructed vertically, and then vitrectomy was performed. Histologic analysis of the incisions was performed. RESULTS: In Group 1, 5 of 9 incisions (55%) demonstrated a two-plane structure. In Group 2, 3 of 9 incisions (33%) demonstrated a two-plane structure. Scleral fibers in the internal aspect of wounds were frayed in all incisions in Group 1 (100%), in 88% of incisions in Group 2, and in all incisions in Group 3 (100%). All incisions in Group 4 were single-planed and two incisions demonstrated disruption on their inner aspect. CONCLUSIONS: Oblique incision architecture was inconsistent within each group. Most incisions showed tissue disruptions in their inner aspect. The internal wound edge disruption reduces the effective distance between the ocular surface and the vitreous cavity. Oblique incisions may not be secure in the immediate postoperative period before wound healing. PMID- 19289987 TI - Long-term follow-up of vitrectomy for diffuse nontractional diabetic macular edema. AB - PURPOSE: To report the long-term results of pars plana vitrectomy for diffuse nontractional diabetic macular edema. DESIGN: Interventional, retrospective, consecutive case series. METHODS: Clinical records of 332 consecutive patients (496 eyes) with diabetic macular edema without a thickened and taut posterior hyaloid on contact lens examination were reviewed. Four hundred eighty-six eyes of 326 consecutive patients were included in this study. All patients underwent pars plana vitrectomy with the creation of a posterior vitreous detachment by one surgeon. Simultaneous phacoemulsification with intraocular lens implantation was performed on 456 phakic eyes. Internal limiting membrane peeling was performed on 178 (36.6%) of 486 eyes. The main outcome measured was best-corrected visual acuity results during follow-up periods. RESULTS: Postoperative follow-up ranged from 12 to 170 months (mean, 74.0 months). Five year follow-up data were available for 356 (71.8%) of 496 eyes. Mean preoperative best-corrected visual acuity significantly increased from 0.19 (20/105) to 0.32 (20/63) at 1 year after surgery (P < 0.0001), and 0.30 (20/67) at the final visit (P < 0.0001). The final best-corrected visual acuity improved in 256 (52.7%) of the 486 eyes, remained unchanged in 152 eyes (31.3%), and worsened in 78 eyes (16.0%). Postoperative major complications included neovascular glaucoma in 19 eyes (3.9%), recurrent vitreous hemorrhage in 10 eyes (2.1%), hard exudate deposits in the center of the macula in 21 eyes (4.2%), and glaucoma in 22 eyes (4.5%). CONCLUSIONS: Pars plana vitrectomy with and without internal limiting membrane peeling appears to be beneficial in eyes with diffuse nontractional diabetic macular edema, and its effectiveness is maintained long term. PMID- 19289988 TI - Pseudodoubling of the optic disk. PMID- 19289989 TI - Diagnostic and therapeutic challenges. PMID- 19289990 TI - Health status of the patients before lung transplantation: a report from Iran. AB - BACKGROUND: Breathing is essential for living. When someone is a lung transplantation candidate, it could be imagined that the breathing difficulties that he/she is experiencing has a great impact on his/her health status. MATERIAL/METHODS: We asked all the patients on the lung transplantation waiting list at Masih Daneshvari Hospital to complete the validated Iranian version of Global Health Questionnaire (GHQ). It was the 28-item version of the questionnaire with scores ranging from 0 to 84. Patients filled the questionnaire while on routine out-patients visits. The higher scores represent greater distress. RESULTS: Seventy patients were requested to participate in the present study out of which 64 filled the questionnaire completely. Mean +/-SD (min-max) scores of the questionnaire subscales were as follows: somatic symptoms =8.2+/ 4.2; anxiety =8.1+/-4.9; 8-14, depression =4+/-4.2; social dysfunction =10.4+/-4 and the overall score =30.6+/-9.5. There was no significant difference in the subscales of the questionnaire based on gender and disease type. Higher age was associated with less social dysfunction (r=-273, p=0.023). Higher level of prednisolone consumption and lower hemoglobin were associated with poorer somatic status (r=0.644, p=0.033; r=-0.410, 0.030 respectively). CONCLUSIONS: Our findings indicate that lung transplantation candidates have extremely poor health state. It seems that pulmonary problems and activity limitations put more pressure on younger patients and causes social difficulties. PMID- 19289991 TI - The early and long term function and survival of kidney allografts stored before transplantation by hypothermic pulsatile perfusion. A prospective randomized study. AB - BACKGROUND: A prospective evaluation of the influence of methods of kidney storage prior to transplantation on long-term graft function has not been shown so far. A retrospective study undertaken in 415 patients in our department showed the benefit of machine perfusion (MP) on long-term results. The aim of the present study was to assess prospectively the long term function and survival of paired kidney allografts retrieved from the same donor, comparing the influence of cold storage (CS) and MP. MATERIAL/METHODS: 74 recipients included in the study received kidneys from 37 cadaveric donors. Kidneys were randomized to storage by CS or MP. There were no significant differences between the groups as to age, gender, duration of ESRD treatment, PRA titres, HLA compatibility and immunosuppressive regimens. RESULTS: At 10 years follow-up recipients of CS stored kidneys returned to dialysis treatment twice as frequently as recipients of MP-stored kidneys (50% vs. 25%, p=0.02). CONCLUSIONS: Kidney storage by MP improves graft survival and reduces the number of patients who return to dialysis treatment at long-term post-transplant. PMID- 19289992 TI - Approach to a target value for 2-hours post dose cyclosporine (C2) during the first week post renal transplantation. AB - BACKGROUND: There is not a wide consensus on whether recommended target ranges for 2-hours post dose cyclosporine (CsA) blood level (C2) are generalizeable to all kidney recipient populations worldwide. In this study we aimed to assess in which C2 level we can obtain the least acute rejection (AR) episodes in our kidney transplanted patients. MATERIAL/METHOD: In a retrospective cross-sectional study, we investigated all our renal recipients with at least a valid C2 blood level at the days between 5-9 post transplantation. All patients were under immunosuppressive therapy with CsA (Neoral), prednisolone and MMF. RESULTS: Hundred forty-four patients were eligible for inclusion in the study. Mean age of the study subjects at the time of transplantation was 36.8+/-16.6 years. 99 (69%) of the patients were male. Overall, 16 (11%) patients experienced AR during the first two weeks post-transplantation. Mean C2 blood levels for patients experiencing AR was 793+/-335 compared with 1028+/-391 for patients without AR (p=0.023). We found that none of the patients with a C2 level of higher than 1300 ng/mL experienced an episode of AR. CONCLUSIONS: According to our findings, we recommend that for minimization purpose of the incident AR episodes among LURD kidney, a C2 blood level of higher than 1300 ng/mL to be obtained during the first week post-transplantation. Alongside, approaching specific C2 targets for patients with different drug regimen or genetic polymorphisms seem necessary. PMID- 19289993 TI - The effect of CYP3A5 and ABCB1 single nucleotide polymorphisms on tacrolimus dose requirements in Caucasian liver transplant patients. AB - BACKGROUND: Tacrolimus is a substrate of cytochrome P-450 (CYP) 3A enzyme and of the drug transporter ABCB1. We have investigated the effects of possible relevant CYP3A5 and ABCB1 single nucleotide polymorphisms (SNPs) present in both donors and recipients on tacrolimus blood levels achieved in a population of 32 Caucasian liver transplant patients. MATERIAL/METHODS: At 1, 3 and 6 months after transplantation, tacrolimus doses (mg/kg/day) and trough blood levels (C(0)) were determined. Polymerase chain reaction followed by restriction fragment length polymorphism analysis was used for genotyping CYP3A5*3 [6986A>G] as well as ABCB1 at exons 21 [2677G>T] and 26 [3435C>T]. RESULTS: 87.5% of the population showed a CYP3A5*3/*3 genotype. For the ABCB1 SNPs, in the case of 3435C>T the total frequency observed for the allelic variant was 50%. For the 2677G>T, the total frequency of the allelic variant was 12.5%, lower than in other Caucasian populations and without any significant linkage with 3435C>T. At 3 and 6 months after transplantation, tacrolimus dose requirements were significantly higher in patients receiving a liver with one copy of the *1 allele compared to those homozygous for the *3 allele (0.111+/-0.057 vs. 0.057+/-0.030 [P<0.05] at 3 month and 0.086+/-0.051 vs. 0.044+/-0.025 [P<0.05] at 6 month). For the recipients' genotypes, the presence of at least one *1 copy tended, though not statistically significantly, to increase tacrolimus doses. With regard to the ABCB1 SNPs, they did not show any influence on tacrolimus dosing requirements. CONCLUSIONS: Pharmacogenetic analysis of CYP3A5 in the donor could contribute to determine the appropriate initial dosage of tacrolimus in liver transplant patients. PMID- 19289994 TI - Does CMV infection increase the incidence of infective endocarditis following kidney transplantation? AB - BACKGROUND: Infective endocarditis (IE) is a rare but life threatening infection after renal transplantation. In addition, coinfection of CMV and IE has not been reported. Therefore, the current study was initiated to determine whether CMV infection is a risk factor for developing of IE after kidney transplantation. MATERIAL/METHODS: In a retrospectively study, we analyzed the medical records of 3700 kidney recipients at two transplant centers in Iran, between January 2000 and June 2008 for infective endocarditis. RESULTS: During the study, 15 patients with IE hospitalized in our centers were included. The predominant causative microorganisms (60%) were group D non-enterococcal streptococci and enterococci. Patient survival rate in all recipients was 66% at 6 months. Data analysis showed no significant differences in 6 months patient survival from hospitalization between both groups with and without CMV infection (P=0.2). The presentation time of infective endocarditis in recipients with CMV coinfection was more likely to be early when compared to CMV negative coinfection patients (P=0.03). CONCLUSIONS: The present study indicates that CMV infection may lead to predispose to infective endocarditis after kidney transplantation. Rapid diagnosis, effective treatment, and prompt recognition of complications in kidney transplant recipients are essential to good patient outcome. PMID- 19289995 TI - Initial hepatosplanchnic blood flow distribution and oxygen metabolism in experimental model of hypotensive brain death. AB - BACKGROUND: Organs from the so-called marginal donors have been used with a significant higher risk of primary non function than organs retrieved from the optimal donors. We investigated the early metabolic changes and blood flow redistribution in splanchnic territory in an experimental model that mimics marginal brain-dead (BD) donor. MATERIAL/METHODS: Ten dogs (21.3+/-0.9 kg), were subjected to a brain death protocol induced by subdural balloon inflation and observed for 30 min thereafter without any additional interventions. Mean arterial and intracranial pressures, heart rate, cardiac output (CO), portal vein and hepatic artery blood flows (PVBF and HABF, ultrasonic flowprobe), and O(2) derived variables were evaluated. RESULTS: An increase in arterial pressure, CO, PVBF and HABF was observed after BD induction. At the end, an intense hypotension with normalization in CO (3.0+/-0.2 vs. 2.8+/-2.8 L/min) and PVBF (687+/-114 vs. 623+/-130 ml/min) was observed, whereas HABF (277+/-33 vs. 134+/-28 ml/min, p<0.005) remained lower than baseline values. CONCLUSIONS: Despite severe hypotension induced by sudden increase of intracranial pressure, the systemic and splanchnic blood flows were partially preserved without signs of severe hypoperfusion (i.e. hyperlactatemia). Additionally, the HABF was mostly negatively affected in this model of marginal BD donor. Our data suggest that not only the cardiac output, but the intrinsic hepatic microcirculatory mechanism plays a role in the hepatic blood flow control after BD. PMID- 19289996 TI - Successful endoscopic and surgical management of non-anastomotic biliary strictures after liver transplantation - case report. AB - BACKGROUND: One of the most relevant biliary complications after liver transplantation are non-anastomotic strictures which occur in about 10-15%. Untreated they lead to cholestasis, severe graft dysfunction, septic complications, secondary cirrhosis and even death. To date they are usually treated by endoscopic or percutaneous placement of stents and balloon dilatation. A significant amount of patients with non-anastomotic strictures require a liver retransplantation. CASE REPORT: A 64 year old patient suffering from HCV induced liver cirrhosis underwent liver transplantation at our hospital. Two months after transplantation due to elevated parameters of cholestasis the patient underwent an endoscopic retrograde cholangiography. Multiple strictures of the bile duct were observed and treated by endoscopic and percutaneous methods until a significant amelioration of the pathological finding in the right liver lobe was achieved. Unfortunately biliary strictures remained in the left liver lobe being resistant to the previous method of treatment. We thus decided to perform a left hemihepatectomy. The postoperative course was unremarkable. CONCLUSIONS: The treatment of our patient consisted of over 25 endoscopic and percutaneous interventions and a left hemihepatectomy. The patient was followed up for two years, during which he had no further complaints being in good health. We demonstrated an example of a successful management of one of the most severe late biliary complications after liver transplantation - the non-anastomotic strictures - avoiding a retransplantation of the organ by endoscopic, percutaneous and surgical intervention. Thus a graft resection seems to be possible. PMID- 19289997 TI - Pulmonary artery stenosis shortly after lung transplantation: successful balloon dilation and stent insertion in one case. AB - BACKGROUND: Pulmonary artery stenosis after lung transplantation is a rare complication. It usually requires surgical correction but even after that the outcome is not favorable.
CASE REPORT: The patient was a 53-years-old woman who was candidate for lung transplantation surgery due to pulmonary fibrosis. After 7 months on waiting list, with severe limitations in daily living activities, she received a single lung transplant in 2007. The surgery was performed without any complication. One day after surgery and after extubation, the patient needed oxygen supplementation through mask with reservoir bag. In bronchoscopy, black-and-white exudate and black membrane that blocked the main bronchus in the transplanted lung was observed. By bronchial lavage the membrane and exudate were successfully removed and patient received antibiotics for documented Aspergillus infection and methylprednisolone pulse therapy for evidences of graft rejection. Despite success in treatments of the mentioned complications, the condition of the patient deteriorated and she became totally dependent to supplemental oxygen. Oxygen consumption level had increase and pulmonary artery pressure was increasing gradually. With suspicion to pulmonary artery stenosis, bronchial CT-Scan with contrast was performed 13 days after transplantation surgery which showed a 50% stenosis. Trans-esophageal echocardiography also showed a stenosis with 40 mmHg gradient. 18 days after transplantation surgery, percutaneous balloon angioplasty was performed which was initially successful but re-stenosis occurred. Seven days later, another balloon angioplasty with stent insertion was performed. After the procedure, the gradient has been removed. Patient was discharged 30 days after transplantation. Follow-up after 10 months revealed no stenosis and the stent was working properly.
CONCLUSIONS: Stent angioplasty can be performed with no problem or complication if pulmonary artery stenosis is seen after lung transplantation.

PMID- 19289998 TI - Channelrhodopsin2 mediated stimulation of synaptic potentials at Drosophila neuromuscular junctions. AB - The Drosophila larval neuromuscular preparation has proven to be a useful tool for studying synaptic physiology. Currently, the only means available to evoke excitatory junctional potentials (EJPs) in this preparation involves the use of suction electrodes. In both research and teaching labs, students often have difficulty maneuvering and manipulating this type of stimulating electrode. In the present work, we show how to remotely stimulate synaptic potentials at the larval NMJ without the use of suction electrodes. By expressing channelrhodopsin2 (ChR2) in Drosophila motor neurons using the GAL4-UAS system, and making minor changes to a basic electrophysiology rig, we were able to reliably evoke EJPs with pulses of blue light. This technique could be of particular use in neurophysiology teaching labs where student rig practice time and resources are limited. PMID- 19289999 TI - Auto-SCT for AML in second remission: CALGB study 9620. AB - We studied the feasibility and efficacy of a two-step approach to Auto-SCT for patients with AML in second remission. Step 1 consisted of consolidation chemotherapy using cytarabine 2000 mg/m(2) i.v. every 12 h for 4 days plus etoposide 40 mg/kg total dose by continuous i.v. infusion over the same 4 days. PBSC were collected after G-CSF stimulation during recovery from this chemotherapy. Step 2, auto-SCT, used a preparative regimen of oral BU 16 mg/kg over 4 days followed by etoposide 60 mg/kg i.v. Of the 50 patients entered on Step 1, two died from treatment complications, and seven failed to proceed to transplantation. A median CD34+ cell dose of 5.9 x 10(6)/kg was collected in a median of three collections. With a median follow-up of 8.2 years, 5-year disease free survival (DFS) is 28%. The most important prognostic factor was cytogenetics, with acute promyelocytic leukemia (APL) patients having a 5-year DFS of 67% compared with 16% for others. We conclude that this two-step approach to autologous transplantation produces good CD34+ mobilization and that this approach has cured some patients. Results in patients with APL are especially promising. PMID- 19290000 TI - Prediction of duration and success rate of unrelated hematopoietic stem cell donor searches based on the patient's HLA-DRB1 allele and DRB1-DQB1 haplotype frequencies. AB - Rapid identification of a matched unrelated donor is essential for patients in need of hematopoietic SCT. We carried out a retrospective evaluation of 549 unrelated donor searches (UDSs), which were completed in 2005 for 23 German transplant centers. On the basis of the patient's HLA-DRB1 allele and DRB1-DQB1 haplotype frequencies, UDSs were divided into four groups with different search success probability predictions. For 90.5% of the patients, an acceptable HLA matched, and for 61.6% an HLA-A-B-Cw-DRB1-DQB1-identical (10/10 matched) unrelated donor was found. The median search duration was 22 days. In the groups with high (n = 318), medium (n = 157), low (n = 56) and very low (n = 18) UDS success probability, an acceptable donor was found for 99.1, 86.6, 75.0 and 22.2% of the patients, and a 10/10-matched donor was found for 78.3, 49.7, 17.9 and 4.5% of the patients, respectively. The median search duration was 20, 27, 45 and 477 days in the groups with high, medium, low and very low probability, respectively. The search success rate and duration can be predicted on the basis of the patient's HLA-DRB1 allele and HLA-DRB1-DQB1 haplotype frequencies. An unrelated donor can be found for most of the patients, even if the indication for transplantation is urgent. PMID- 19290001 TI - Cervical fascial and muscular involvement in chronic GVHD after allo-SCT and radiation therapy. PMID- 19290002 TI - Criteria for initiation and evaluation of minority donor programs and application to the example of donors of Turkish descent in Germany. AB - Minority donor programs aim to improve access to unrelated hematopoietic SCT for specific ethnic groups through directed donor recruitment. We have developed criteria for initiation and evaluation of such programs and applied them to the situation of donors of Turkish descent in Germany, as well as a program by DKMS German Bone Marrow Center that targets this group. Criteria for program initiation include the number of accessible minority donors, potential impact on the chances of finding matching donors, and general access to unrelated transplantation for patients of the targeted group. Success criteria comprise number and availability of recruited donors, the effect of these donors on the HLA phenotype distribution of a donor file, and the number of donations resulting from the program. More than 40 000 donors of Turkish descent have been recruited within the analyzed program to date. Recruited minority donors show more favorable demographic characteristics but lower availability rates than do German donors. Although HLA haplotype distributions of Turkish and German donors differ considerably, patients with common Turkish HLA phenotypes should benefit from the German donor pool even without a specific minority program. The analysis of donations from minority donors, however, shows specific benefits for patients with rare HLA phenotypes. PMID- 19290003 TI - Utility of the psychosocial assessment of candidates for transplantation (PACT) scale in allogeneic BMT. AB - The psychosocial assessment of candidates for transplantation (PACT) scale was completed before the transplant on 120 patients who underwent allogeneic transplant from November 2003 to June 2007. The PACT has eight subscales, each rated on a 5-point scale, and an initial and final rating independently based on the rater's overall impressions of the candidate's acceptability for transplant. This exploratory study assessed the clinical utility of the PACT scale for psychosocial screening in allogeneic BMT. Associations of the PACT subscales and the final rating with sixteen post transplant medical outcomes were examined using the Jonchkheere-Terpstra test, the Cochran-Armitage test or the Cox proportional hazards analysis. Significant relationships (PT (rs4252041, minor allele frequency=4%) SNP was associated with the primary outcome total fat mass (P=0.003) and regional fat masses, but not with lean body mass or serum IL-1 receptor 1 (IL1RN) levels. This SNP was also associated with body fat when correcting the earlier reported IL1RN+2018 T>C (rs419598) SNP (in linkage disequilibrium with a well-studied variable number tandem repeat of 86 bp). The association between rs4252041 SNP and body fat was confirmed in the older MrOS population (P=0.03). The rs4252041 SNP was part of three haplotypes consisting of five adjacent SNPs that were identified by a sliding window approach. These haplotypes had a highly significant global association with total body fat (P<0.001). None of the other investigated members of the IL-1 gene family displayed any SNPs that have not been described previously to be significantly associated with body fat. CONCLUSIONS: The IL1RN gene, shown to enhance obesity by suppressing IL-1 effects in experimental animals, have not [corrected] previously described gene polymorphisms and haplotypes that are associated with fat, but not lean mass in two populations of men. PMID- 19290010 TI - A 4-year, cluster-randomized, controlled childhood obesity prevention study: STOPP. AB - OBJECTIVE: To assess the efficacy of a school-based intervention programme to reduce the prevalence of overweight in 6 to 10-year-old children. DESIGN: Cluster randomized, controlled study. SUBJECTS: A total of 3135 boys and girls in grades 1-4 were included in the study. METHODS: Ten schools were selected in Stockholm county area and randomized to intervention (n=5) and control (n=5) schools. Low fat dairy products and whole-grain bread were promoted and all sweets and sweetened drinks were eliminated in intervention schools. Physical activity (PA) was aimed to increase by 30 min day(-1) during school time and sedentary behaviour restricted during after school care time. PA was measured by accelerometry. Eating habits at home were assessed by parental report. Eating disorders were evaluated by self-report. RESULTS: The prevalence of overweight and obesity decreased by 3.2% (from 20.3 to 17.1) in intervention schools compared with an increase of 2.8% (from 16.1 to 18.9) in control schools (P<0.05). The results showed no difference between intervention and controls, after cluster adjustment, in the longitudinal analysis of BMIsds changes. However, a larger proportion of the children who were initially overweight reached normal weight in the intervention group (14%) compared with the control group (7.5%), P=0.017. PA did not differ between intervention and control schools after cluster adjustment. Eating habits at home were found to be healthier among families with children in intervention schools at the end of the intervention. There was no difference between children in intervention and control schools in self-reported eating disorders. CONCLUSIONS: A school-based intervention can reduce the prevalence of overweight and obesity in 6 to 10-year-old children and may affect eating habits at home. The effect of the intervention was possibly due to its effect on healthy eating habits at school and at home rather than on increased levels of PA. PMID- 19290011 TI - Medical care of overweight children under real-life conditions: the German BZgA observation study. AB - OBJECTIVE: Current care for overweight children is controversial, and only few data are available concerning the process of care, as well as the outcome under real-life conditions. METHODS: A nationwide survey of treatment programs for overweight children and adolescents in Germany identified 480 treatment centers. From 135 institutions that had agreed to participate in this study of process of care and outcome, 48 randomly chosen institutions were included in the study. All 1916 overweight children (mean age 12.6 years, 57% female, mean body mass index 30.0 kg/m(2)), who presented at these institutions for lifestyle interventions, were included in this study. Diagnostic procedures according to guidelines and effect of lifestyle interventions on weight status at end of treatment were analyzed. RESULTS: Children treated <3 months were older and more obese, whereas children with >3 months treatment duration demonstrated more cardiovascular risk factors at baseline. On the basis of an intention-to-treat analysis, 75% of the children reduced their overweight. The reduction of overweight varied widely between the treatment institutions (intracluster correlation coefficient 0.15 in the multiple regression model reflecting the intracenter correlation). Screening for hypertension, disturbed glucose metabolism and dyslipidemia was performed in 52% of the children at baseline and in 10% at the end of intervention. CONCLUSION: Overweight reduction is achievable with lifestyle intervention in clinical practice. However, because the clientele, treatment approach and outcome varied widely between different institutions, and screening for comorbidities was seldomly performed as recommended, quality criteria for institutions have to be implemented to improve medical care of overweight children under real-life conditions. PMID- 19290012 TI - Serum resistin correlates with central obesity but weakly with insulin resistance in Chinese children and adolescents. AB - OBJECTIVE: Resistin has been linked with obesity and hypothesized as a potential marker of insulin resistance in addition to being linked with acute inflammation. However, these links are still highly controversial in humans. Our goal was to examine resistin levels in relation to obesity, insulin resistance and inflammation markers in a large population of Asian children and adolescents. METHODS: Children and adolescents (n=3472) aged 6-18 years, boys (n=1765) and girls (n=1707), were assessed for body size parameters, pubertal development, blood lipids, glucose, insulin, resistin, C-reactive protein (CRP), adiponectin and complement C3 (C3) levels. RESULTS: Resistin increased with central obesity in both genders but not with simple adiposity in boys. Several markers associated with central obesity correlated in a gender-specific fashion with plasma resistin. Waist circumference, fat-mass percentage, waist-to-height ratio and body mass index (BMI) positively correlated with resistin in both genders. Blood lipids such as triglycerides, nonesterified fatty acids (NEFA) and low-density lipoprotein cholesterol, diastolic and systolic blood pressure correlated positively with resistin in boys. NEFA, high-density lipoprotein cholesterol (negatively) and inflammation markers, such as CRP and C3, positively correlated with resistin in girls. There was no correlation between resistin and adiponectin, and no association of adiponectin with resistin quintiles in either boys or girls. In both boys and girls, resistin tended to decrease with age, with girls having higher levels than boys. Few indices of insulin resistance were linked with plasma resistin in either gender. CONCLUSION: In this population, plasma resistin levels are a weak biochemical marker of metabolic dysfunction defined by central obesity, adiposity and inflammation and does not predict insulin resistance. Only a small proportion of resistin variation can be explained by factors related to metabolic syndrome, suggesting that resistin is not strongly implicated in a concentration-dependent fashion in any of the examined pathologies. PMID- 19290013 TI - The efficacy of alpha-tocopherol in functional recovery of spinal cord injured rats: an experimental study. AB - OBJECTIVE: The objective of this study is to examine the effects of the alpha tocopherol on rats with spinal cord injury (SCI). SETTING: Research Center, Sultan Bin Abdulaziz Humanitarian City, Riyadh, Kingdom of Saudi Arabia. METHOD: Female Sprague-Dawley rats weighing 180-220 g were anesthetized with chloral hydrate (450 mg kg(-1) body weight) by intraperitoneal injection and laminectomy was performed at the T 7-8 level leaving the dura intact. A compression plate (2.2 x 5.0 mm) was loaded with a weight of 35 g placed on the exposed cord for 5 min to create SCI. The subjects were divided into three groups of eight rats each. Group 1 served as control (SCI+saline); whereas groups 2 and 3 served as test groups, alpha-tocopherol was given orally in doses of 1000 mg kg(-1) body weight for group 2 and 2000 mg kg(-1) body weight for group 3, respectively. Daily activities were recorded in the activity cage for 14 days post-operatively. RESULTS: At day 1 (baseline, 24 h after the surgery), there was no significant difference between mean motor scores of all groups. After day 1, the three groups showed continuous improvement in motor score; such improvement was maintained throughout the duration of the study with different levels for each group. By the end of the study (day 14), groups 2 and 3 showed statistically significant improvement in the mean motor score compared with group 1 (P<0.05). However, no significant difference was observed between test groups 2 and 3 by the end of the study. CONCLUSION: The results suggest that the administration of alpha tocopherol may have reparative effects for SCI because of its antioxidant effect. PMID- 19290014 TI - Body composition assessment in athletes with spinal cord injury: comparison of field methods with dual-energy X-ray absorptiometry. AB - STUDY DESIGN: Cross-sectional. OBJECTIVES: To compare relative body fatness (%Fat) estimates from field methods (skinfold thickness measurement (SKF) and bioelectrical impedance analysis (BIA)) with measures by dual-energy X-ray absorptiometry (DXA). SETTING: University of Illinois, Urbana-Champaign, IL, USA. METHODS: Field methods used both three- and seven-site SKF prediction equations and BIA generalized, spinal cord injury (SCI)-specific and athlete-specific equations. DXA was used as the reference method. College-aged varsity athletes with SCI (women=8, men=8; time since injury 16.2+/-5.7 years; injury level range T5-L5) were recruited. RESULTS: Mean BMI was 20.8+/-2.6 and 22.5+/-2.1 kg m(-2), and mean DXA %Fat was 31.9+/-3.8 and 20.6+/-8.4%, for women and men, respectively. All field methods under-predicted the %Fat when compared with DXA (ranges in mean differences: SKF women 2.9-8.2%, SKF men 6.9-12.4%; BIA women 0.5 3.9%, BIA men 0.3-7.0%). None of the field methods accurately predicted the %Fat compared with DXA (total error (TE): SKF women 7.4-12.1%, SKF men 8.4-15.2%; BIA women 5.1-9.3%, BIA men 6.7-10.7%). Of the SKF and BIA prediction equations, Evans et al.'s three-site SKF (r=0.95, P<0.001, standard error of the estimate (SEE)=2.8 %Fat) prediction equation provided the best fit for this population. CONCLUSION: Further studies with larger samples are necessary to develop appropriate prediction equations for field methods in the athletic SCI population. PMID- 19290015 TI - Natural killer cells: NK cells stroll down the memory lane. PMID- 19290016 TI - Allogeneic stromal cell implantation in brain tissue leads to robust microglial activation. AB - Although adult and embryonic stem cell-based therapy for central nervous system (CNS) injury is being developed worldwide, less attention is given to the immunological aspects of allogeneic cell implantation in the CNS. The latter is of major importance because, from a practical point of view, future stem cell based therapy for CNS injury will likely be performed using well-characterised allogeneic stem cell populations. In this study, we aimed to further describe the immunological mechanism leading to rejection of allogeneic bone marrow-derived stromal cells (BM-SC) after implantation in murine CNS. For this, we first investigated the impact of autologous and allogeneic BM-SC on microglia activation in vitro. Although the results indicate that both autologous and allogeneic BM-SC do not activate microglia themselves in vitro, they also do not inhibit activation of microglia after exogenous stimuli in vitro. Next, we investigated the impact of allogeneic BM-SC on microglia activation in vivo. In contrast to the in vitro observations, microglia become highly activated in vivo after implantation of allogeneic BM-SC in the CNS of immune-competent mice. Moreover, our results suggest that microglia, rather than T-cells, are the major contributors to allograft rejection in the CNS. PMID- 19290017 TI - Modelling naive T-cell homeostasis: consequences of heritable cellular lifespan during ageing. AB - Within an individual, the population of mature naive T cells is maintained throughout life by both input from the thymus and homeostatic proliferation in the periphery. Here, we develop a mathematical model of this process of naive T cell homeostasis, and use it to explore questions of lifespan, inheritance and receptor repertoire during ageing. By assuming lifespan is largely determined by a heritable trait reset on mitosis, we show that homeostatic proliferation leads naturally to a longer lived population with age. A plausible candidate for the heritable trait influencing lifespan is T-cell receptor affinity for major histocompatibility molecules loaded with self-peptides. Concurrently with increasing lifespan, receptor diversity decreases with age, thus quantitatively linking these two phenomena. These results depend on the thymus involuting with age so that homeostatic proliferation becomes the dominant mode of replacement of the naive T-cell repertoire. PMID- 19290018 TI - Association of nicotinic acetylcholine receptor subunit alpha 4 polymorphisms with nicotine dependence in 5500 Germans. AB - Polymorphisms in the CHRNA4 gene coding the nicotinic acetylcholine receptor subunit alpha 4 have recently been suggested to play a role in the determination of smoking-related phenotypes. To examine this hypothesis, we conducted a genetic association study in three large samples from the German general population (N(1)=1412; N(2)=1855; N(3)=2294). Five single-nucleotide polymorphisms in CHRNA4 were genotyped in 5561 participants, including 2707 heavily smoking cases (regularly smoking at least 20 cigarettes per day) and 2399 never-smoking controls (or=0.4) lower on sexual domains, and statistically (P<0.05) lower on physical function and vitality. Estimated free testosterone and measured serum estradiol had increased from baseline in most patients, but did not correlate with HRQoL outcomes 5 years after the start of treatment. PMID- 19290020 TI - A partition-ligation-combination-subdivision EM algorithm for haplotype inference with multiallelic markers: update of the SHEsis (http://analysis.bio-x.cn). PMID- 19290021 TI - Novel function of perforin in negatively regulating CD4(+) T cell activation by affecting calcium signaling. AB - Perforin is a pore-forming protein engaged mainly in mediating target T cell death and is employed by cytotoxic T lymphocytes (CTLs) and natural killer cells. However, whether it also plays a role in conventional CD4(+) T cell function remains unclear. Here we report that in perforin-deficient (PKO) mice, CD4(+) T cells are hyperproliferative in response to T cell receptor (TCR) stimulation. This feature of hyperproliferation is accompanied by the enhancement both in cell division and in IL-2 secretion. It seems that the perforin deficiency does not influence T cell development in thymus spleen and lymph node. In vivo, perforin deficiency results in increased antigen-specific T cell proliferation and antibody production. Furthermore, PKO mice are more susceptible to experimental autoimmune uveitis. To address the molecular mechanism, we found that after TCR stimulation, CD4(+) T cells from PKO mice display an increased intracellular calcium flux and subsequently enhance activation of transcription factor NFAT1. Our results indicate that perforin plays a negative role in regulating CD4(+) T cell activation and immune response by affecting TCR-dependent Ca(2+) signaling. PMID- 19290022 TI - Differential distributions and trafficking properties of dopamine D1 and D5 receptors in nerve cells. AB - OBJECTIVE: To explore the possible differential trafficking properties of the dopamine D1-like receptor subtypes, D1 receptor and D5 receptor. METHODS: To visualize distributions of dopamine D1-like receptor subtypes at subcellular level, the yellow and cyan variants of green fluorescent protein (GFP) were used to tag D1 and D5 receptors. After transfection with the tagged dopamine receptors, the neuroblastoma cells NG108-15 were treated with D1 agonist SKF38393 or acetylcholine (ACh). Then we observed the subcellular distributions of the tagged receptors under the confocal microscopy and tried to determine trafficking properties by comparing their distribution patterns before and after the drug treatment. RESULTS: In resting conditions, D1 receptors located in the plasma membrane of NG108-15 cells, while D5 receptors located in both plasma membrane and cytosol. With the pre-treatment of SKF38393, the subcellular distribution of D1 receptors was changed. The yellow particle-like fluorescence of tagged D1 receptors appeared in the cytosol, indicating that D1 receptors were internalized into cytosol from the cell surface. Same situation also occurred in ACh pre treatment. In contrast, the subcellular distribution of D5 receptors was not changed after SKF38393 or ACh treatment, indicating that D5R was not translocated to cell surface. Interestingly, when D1 and D5 receptors were co-expressed in the same cell, both kept their distinct subcellular distribution patterns and the trafficking properties. CONCLUSION: Our present study reveals that in NG108-15 nerve cells, dopamine D1 and D5 receptors exhibit differential subcellular distribution patterns, and only D1 receptor has a marked trafficking response to the drug stimulation. We further discuss the potential role of the differential trafficking properties of D1-like receptors in complex modulation of DA signaling. PMID- 19290023 TI - Preparation and application of rat myostatin antiserum. AB - OBJECTIVE: To prepare and identify a polyclonal antibody against rat myostatin and investigate myostatin expression in the rat atrophic gastrocnemius muscle after tibial nerve crush. METHODS: The purified fusion protein was used as antigen to immunize rabbits for the preparation of polyclonal antibody. The polyclonal antibody of the protein was measured by enzyme linked immunosorbent assay (ELISA), western-blot and immunochemistry. Myostatin protein expression levels in normal and atrophic gastrocnemius muscle were detected by western-blot and immunochemistry assays. RESULTS: The GST-myostatin had a purity of 96% and possessed high titer and specificity. The level of myostatin in gastrocnemius muscle significantly increased one week after tibial nerve crush, reached the peak on day 14, and then returned to normal level on day 28. CONCLUSION: We have successfully made antiserum of rat myostatin and found that the expression level of myostatin protein in the gastrocnemius after tibial nerve crush-induced atrophy was time-dependent. This study provides an experimental basis to clarify the possible role of myostatin during skeletal muscle atrophy. PMID- 19290024 TI - Permanent myopathy caused by mutation of SCN4A Metl592Val: Observation on myogenesis in vitro and on effect of basic fibroblast growth factor on the muscle. AB - OBJECTIVE: The present study is to observe in vitro the proliferation ability of the muscle cells from permanent myopathy (PM) patients of nomokalaemic periodic paralysis (normKPP), which is caused by mutations of Met1592Val in the skeletal muscle voltage gated sodium channel (SCN4A) gene on chromosome 17q23.1. We also evaluate the possible effect of the foreign basic fibroblast growth factor (bFGF) in preventing and curing PM. METHODS: The gastrocnemius muscle cells were taken from two male patients with PM of the same Chinese family with Met1592Val mutation of SCN4A, determined by gene screening. Four male patients suffering from the skeletal injury without PM were taken as control. All preparations were protogenerationally cultured in vitro. Proliferation of the cultured preparations was measured by MTT. Activities of the lactic dehydrogenase (LDH), creatine kinase (CK), and protein content in these cells were also detected. The effects of bFGF with different doses (10 ng/mL, 20 ng/mL, 40 ng/mL, 80 ng/mL, 120 ng/mL and 160 ng/mL) on the above mentioned parameters were also evaluated. RESULTS: Cells from both PM and control subjects were successfully cultured in vitro. The cultivation of the muscle cells from PM patients in vitro was not yet seen. Results indicated the obvious stimulation of bFGF on cell proliferation, activities of LDH and CK, protein synthesis, in a dose dependent manner. The optimal dose of bFGF was 120 ng/mL (P<0.05), beyond which greater dose caused a less effect. The effect of bFGF on 160 ng /mL was stronger than that on 80 ng/mL, but there was no significant difference (P>0.05). CONCLUSION: Myoblastic cells from patients with PM had a weaker ability of developing into the myotubules, thus they were unable to perform effective regeneration, which resulted in a progressive necrosis. The exogenous bFGF could promote the division and proliferation of the muscle cells in vitro. These results shield a light on bFGFos potential role in preventing and treating PM. PMID- 19290025 TI - Protective effects of edaravone against cobalt chloride-induced apoptosis in PC12 cells. AB - OBJECTIVE: To investigate the neuroprotective effects of edaravone (Eda) on cobalt chloride (CoCl2)-induced oxidative stress and apoptosis in cultured PC12 cells as well as the underlying mechanisms. METHODS: PC12 cells impaired by CoCl2 were used as the cell model of hypoxia. MTT (methyl thiazolyl tetrazolium) was used to assay the viability of the PC12 cells exposed to Eda with gradient concentrations; Hochest 33258 stain assay was used to analyze the apoptosis ratio of the PC12 cells; Bcl-2 and Bax protein levels in PC12 cells were examined by western blotting. ROS level, the mitochondrial transmembrane potential and caspase-3 activity in each group were detected by spectrofluorometer. RESULTS: CoCl2 treatment caused the loss of cell viability in PC12 cells, which was associated with the elevation of apoptotic rate, the formation of ROS and the disruption of mitochondrial transmembrane potential. CoCl2 also significantly induced the upregulation of Bax/Bcl-2 ratio and the activation of caspase-3. In contrast, Eda significantly reversed these phenotypes, with its maximum protective effect at 0.1 micromol/L. CONCLUSION: These results indicated that Eda could protect PC12 cells from CoCl2-induced cytotoxicity, and this protection might be ascribed to its anti-oxidative and anti-apoptotic activities. PMID- 19290026 TI - HLB-1 functions as a new regulator for the organization and function of neuromuscular junctions in nematode Caenorhabditis elegans. AB - OBJECTIVE: To study the role of HLB-1 in regulating the organization and function of neuromuscular junctions in nematode Caenorhabditis elegans. METHODS: To evaluate the functions of HLB-1 in regulating the organization and function of neuromuscular junctions, effects of hlb-1 mutation on the synaptic structures were revealed by uncovering the expression patterns of SNB-1::GFP and UNC 49::GFP, and pharmacologic assays with aldicarb and levamisole were also used to test the synaptic functions. Further rescue and mosaic analysis confirmed HLB-1's role in regulating the organization and function of neuromuscular junctions. RESULTS: Loss of HLB-1 function did not result in defects in neuronal outgrowth or neuronal loss, but caused obvious defects of SNB-1::GFP and UNC-49::GFP puncta localization, suggesting the altered presynaptic and postsynaptic structures. The mutant animals exhibited severe defects in locomotion behaviors and altered responses to an inhibitor of acetylcholinesterase and a cholinergic agonist, indicating the altered presynaptic and postsynaptic functions. Rescue and mosaic analysis experiments suggested that HLB-1 regulated synaptic functions in a cell nonautonomously way. Moreover, HLB-1 expression was not required for the presynaptic active zone morphology. Genetic evidence further demonstrated that hlb-1 acted in a parallel pathway with syd-2 to regulate the synaptic functions. CONCLUSION: HLB-1 appeared as a new regulator for the organization and function of neuromuscular junctions in C. elegans. PMID- 19290027 TI - No spatial memory deficit exists in Kunming mice that recently recovered from motor defects following 3-nitropropionic acid intoxication. AB - OBJECTIVE: Numerous studies have described both motor defects and cognitive impairments in several strains of rodents following 3-nitropropionic acid (3-NP) intoxication. In the present study, we investigated spatial recognition memory in Kunming mice that just recovered from motor defects induced by 3-NP. METHODS: Mouse model was made by systemic subacute 3-NP treatment, and spatial recognition memory was measured through the Y-maze Test, a simple two-trial recognition test. RESULTS: (1) On day 15 following 3-NP treatment, affected Kunming mice did not show motor defects in the Rotarod test and presented normal gait again. (2) In the following Y-maze test after 1h interval, the percentage (90.0%) of mice showing novel arm preference in 3-NP treatment group was significantly higher than the random chance level (50%), although it was only slightly higher than that (83.3%) in control group. On day 45 after 3-NP treatment, mice failed to choose unfamiliar novel arm as first choice, and the same occured in the control group. (3) For both post-intoxicated (on day 15 and day 45 following 3-NP treatment) and control groups, the duration in the novel arm and the frequency of entering it, were longer and higher compared with familiar start and other arms. For these mice that recently recovered from motor defects following 3-NP intoxication, no spatial memory deficits were observed through Y-maze Test. CONCLUSION: Kunming mice used in our assays might possess resistance to cognitive impairment induced by 3-NP, which is consistent with previous findings in Swiss EPM-M1 mice. PMID- 19290028 TI - A review on research progress of transketolase. AB - Transketolase (TK), a thiamine diphosphate (ThDP)-dependent enzyme, catalyzes several key reactions of non-oxidative branch of pentose phosphate pathway. TK is a homodimer with two active sites that locate at the interface between the contacting monomers. Both ThDP and bivalent cations are strictly needed for TK activation, just like that for all ThDP-dependent enzymes. TK exists in all organisms that have been investigated. Up to now, one TK gene (TKT) and two transketolase-like genes (TKTL1 and TKTL2) have been identified in human genome. TKTL1 is reported to play a pivotal role in carcinogenesis and may have important implications in the nutrition and future treatment of patients with cancer. Researchers have found TK variants and reduced activities of TK enzyme in patients with neurodegenerative diseases, diabetes, and cancer. Recent studies indicated TK as a novel role in the prevention and therapy of these diseases. PMID- 19290029 TI - Castleman's Disease. A Review. AB - Castleman's disease is characterized by a non-clonal lymph node hyperplasia. Pathologically it is classified as hyaline vascular, plasmacytic, or mixed cellularity types, and clinically it may adopt a unicentric (localized) or multicentric presentation. An association of the disease with HIV infection has been found. Many uncertainties remain concerning the etiopathogenesis and the optimal treatment of this rare condition. PMID- 19290030 TI - HIV/human T-cell lymphotropic virus coinfection revisited: impact on AIDS progression. AB - Human T-cell lymphotropic viruses type 1 and 2 are retroviruses that share the same routes of transmission as HIV-1. Since these agents are prevalent simultaneously in different parts of the world, coinfection is a frequently reported event. However, prevalence rates of coinfection differ for distinct populations and regions of the world or for each virus, with human T-cell lymphotropic virus type 1 being more prevalent among HIV-1-infected individuals in the Southern hemisphere, while type 2 is more frequently found in the Northern hemisphere. In common, they share the tropism for T-lymphocytes, although human T cell lymphotropic virus type 1 and HIV-1 are predominantly CD4+ T-cell tropic and human T-cell lymphotropic virus type 2 preferentially infects CD8+ cells. The biological properties of HIV-1 are distinct of those found in human T-cell lymphotropic virus 1/2. This fact makes possible an in vivo interaction between these agents, when coinfecting the same patients, with potentially relevant clinical implications. The available evidence suggests a protective role for coinfection by human T-cell lymphotropic virus type 2 on AIDS progression. This hypothesis is supported by several laboratory evidences, as well as by a number of clinical studies that found no significant interaction between human T-cell lymphotropic virus type 2 and HIV-1, or even detected a protective effect on HIV 1 disease. On the other hand, human T-cell lymphotropic virus type 1 seems to be a significant cofactor, with a potentially important role in HIV-1 infection. Although the clinical evidence is still controversial with regard to the real impact that coinfection exerts on clinical evolution, the majority of studies suggest it is associated with a modification of the natural history of HIV-1 infection, with a faster clinical progression and a shorter survival time. The main limitation of the available data is due to methodological problems in the majority of studies, which weaken the validity of their conclusions. A common finding in coinfection by both human T-cell lymphotropic virus type 1 and 2 is the increase in CD4+ cell count, but without any additional immune benefit for patients. Due to the limited available data, we need more, larger studies, designed to respond to the pending questions on the real significance of coinfection by these retroviruses. PMID- 19290031 TI - Characterization and structural analysis of HIV-1 integrase conservation. AB - The HIV-1 integrase, responsible for the chromosomal integration of the newly synthesized double-stranded viral DNA into the host genomic DNA, represents a new and important target of potential clinical relevance. For instance, two integrase inhibitors, raltegravir and elvitegravir, have been shown to be promising in clinical trials, and the first has been recently made available for clinical practice. As is the case for other antiviral drugs, drug resistance to integrase inhibitors occurs both in vitro and/or in vivo through the selection of mutations within the HIV genome. Indeed, many integrase mutations have already been associated with resistance to all the different integrase inhibitors tested in in vitro and/or in vivo studies. Among them, about 40 substitutions have been specifically associated with the development of resistance to raltegravir and/or elvitegravir; some of them were also found in vivo in patients failing such integrase inhibitors. The relevance of integrase mutations in clinical practice has yet to be defined, in light of the lack of long-term follow-up of treated patients and the limited data about the prevalence of integrase inhibitor associated mutations in integrase inhibitor-naive patients (either untreated, or treated with antiretrovirals not containing integrase inhibitors). Therefore, by structural analysis elaboration and literature discussion, the aim of this review is to characterize the conserved residues and regions of HIV-1 integrase and the prevalence of mutations associated with integrase inhibitor resistance, by matching data originated from a well-defined cohort of HIV-1 B subtype-infected individuals (untreated and antiretroviral-treated) and data originated from the public Los Alamos Database available in the literature (all patients integrase inhibitor-naive by definition). In integrase inhibitor-naive patients, 180 out of 288 HIV-1 integrase residues (62.5%) are conserved (< 1% variability). Residues involved in protein stability, multimerization, DNA binding, catalytic activity, and in the binding with the human cellular cofactor LEDGF/p75 are fully conserved. Some of these residues clustered into large defined regions of consecutive invariant amino acids, suggesting that consecutive residues in specific structural domains are required for the correct performance of HIV-1 integrase functions. All primary signature mutations emerging in patients failing raltegravir (Y143R, Q148H/K/R, N155H) or elvitegravir (T66I, E92Q, S147G, Q148H/K/R, N155H), as well as secondary mutations (H51Y, T66A/K, E138K, G140S/A/C, Y143C/H, K160N, R166S, E170A, S230R, D232N, R263K) were completely absent or highly infrequent (< 0.5%) in integrase inhibitor-naive patients, either infected with HIV-1 B subtype (drug-naive or antiretroviral-treated), or non-B subtypes/group N and O. Differently, other mutations (L74M, T97A, S119G/R, V151I, K156N, E157Q, G163K/R, V165I, I203M, T206S, S230N) occurred as natural polymorphisms with a different prevalence according to different HIV-1 subtype/circulating recombinant form/group. In conclusion, the HIV-1 integrase in vivo is an enzyme requiring the full preservation of almost two-thirds of its amino acids in the absence of specific integrase inhibitor pressure. Primary mutations associated with resistance to integrase inhibitors clinically relevant today are absent or highly infrequent in integrase inhibitor-naive patients. The characterization of the highly conserved residues (involved in protein stability, multimerization, DNA binding, catalytic activity, LEDGF binding, and some with still poorly understood function) could help in the rational design of new HIV-1 inhibitors with alternative mechanisms of action and more favorable resistance profiles. PMID- 19290032 TI - Risk factors for gastrointestinal adverse events in HIV treated and untreated patients. AB - Advanced immunosuppression from HIV infection can lead to gastrointestinal symptoms such as diarrhea, nausea, vomiting, dysphagia, weight loss, and abdominal pain. There is a complex, combined effect of HIV infection plus antiretroviral treatment on the incidence of gastrointestinal symptoms, and, for some trials, the majority of gastrointestinal adverse events may not be related to antiretroviral treatment. Antiretroviral treatment can lead to improvements in gastrointestinal symptoms for patients with advanced immunosuppression. This was observed in the TORO trials of enfuvirtide and the DUET trials of etravirine, which were conducted in highly treatment experienced patients with low baseline CD4 counts. While antiretroviral treatment can improve immune function, leading to fewer gastrointestinal symptoms, this could be counter-balanced by adverse gastrointestinal toxicity profiles from certain antiretrovirals. Ritonavir boosted protease inhibitors show a range of gastrointestinal side effects; there are differences in tolerability within this class of antiretrovirals, influenced both by the dose of ritonavir used and the choice of boosted protease inhibitor. Overall, lopinavir/ritonavir and fosamprenavir/ritonavir tend to show the highest rates of drug-related grade 2-4 diarrhea, compared with atazanavir/ritonavir, darunavir/ritonavir, or saquinavir/ritonavir. Of the nucleoside analogs, zidovudine leads to a well-characterized problem of nausea. Issues relating to gastrointestinal complications are often subjective, reliant upon patient reporting and perception, along with clinician interaction and intervention. In trial publications, many different systems are used to present gastrointestinal adverse events. Most are based on the US Division of AIDS Grading Scale, ranging from grade 1 (mild) to grade 4 (life-threatening). Clinical trials most commonly report grade 2-4 gastrointestinal adverse events, which are at least possibly related to study medication. In future, it is important for clinical trials to report gastrointestinal adverse events in a consistent way. The percentage of patients with drug-related grade 2-4 events should be reported. In addition, the percentage with any grade 2-4 gastrointestinal adverse event should be included, since there could be subjectivity in the assessment of drug relatedness in open label clinical trials. The percentage of patients who use medications to lessen the symptoms of diarrhea and other gastrointestinal adverse events should also be reported. PMID- 19290033 TI - HIV-1 genotypic drug resistance interpretation rules - 2009 Spanish guidelines. AB - Interpreting the results of drug resistance tests for HIV-1 is one of the most difficult tasks for both clinicians and virologists. There are many amino acid changes in viral proteins influencing the susceptibility to specific drugs, causing loss of activity or conversely hypersusceptibility. Moreover, the results of interactions derived from complex mutational patterns are difficult to predict. Different interpretation algorithms have been developed to facilitate the translation of information obtained in the genotypes to clinicians. Controversy exists, however, regarding the impact of genotypic changes over the activity of many antiretroviral drugs. Based on virologic outcomes, scientific literature, and expert opinion, the Drug Resistance Platform of the Spanish AIDS Research Network (RIS, Red de Investigacion en SIDA) has developed over the last years its own interpretation system. Herein, we present the 2009 guidelines, in which special efforts have been made to standardize the criteria for interpreting resistance mutations for compounds within the same drug family and to facilitate the clinical interpretation of HIV-1 resistance genotypes. PMID- 19290036 TI - Multicentre inter-cohort studies - reliable responses for HIV disease management? AB - The most recent evidences on HIV infection were obtained through large multicentre trials/inter-cohort analyses including patients enrolled by hundreds of investigators and co-investigators around the world. While extensively pooled data are needed to assess infrequent events (rare toxicities), long-term endpoints, or safety (e.g. mortality or lipodystrophy syndrome), or to perform comparative studies between different treatments, relevant biases/distortions descending from the extremely elevated number of enrolling centers/investigators are expected, while the reports among quoted co-authors of all investigators/co investigators supports an unacceptable number of presumptive authors. As is known, a relevant number of outstanding multicentre/inter-cohort studies signed by thousands of co-authors, comprehensively quoted in all bibliographic databases, have been published in the last months by leading medical journals (starting just from 2008), especially in the attempt to focus on some infrequent HIV disease complications, and regarding efficacy and tolerability of novel antiretroviral therapies (ART). Some representative studies are summarized in the table. PMID- 19290037 TI - Protein Binding to Peptide-Imprinted Porous Silica Scaffolds. AB - Of the many types of biomolecules used for molecular imprinting applications, proteins are some of the most useful, yet challenging, templates to work with. One method, termed the 'epitope approach', involves imprinting a short peptide fragment of the protein into the polymer to promote specific adsorption of the entire protein, similar to the way an antigen binds to an antibody via the epitope. Whole lysozyme or the 16 residue lysozyme C peptide was imprinted into porous silica scaffolds using sol-gel processing. After removing template, scaffolds were exposed to lysozyme and/or RNase A, which was used as a competitor molecule of comparable size. When comparing protein- to peptide-imprinted scaffolds, similar amounts of lysozyme and RNase were bound from single protein solutions. However, while whole lysozyme-imprinted scaffolds showed about 4:1 preferential binding of lysozyme to RNase, peptide-imprinted scaffolds failed to show statistical significance, even though a slight preferential binding trend was present. These initial studies suggest there is potential for using peptide imprinting to create specific protein-binding sites on porous inorganic surfaces, although further development of the materials is needed. PMID- 19290038 TI - The population structure of Glossina palpalis gambiensis from island and continental locations in Coastal Guinea. AB - BACKGROUND: We undertook a population genetics analysis of the tsetse fly Glossina palpalis gambiensis, a major vector of sleeping sickness in West Africa, using microsatellite and mitochondrial DNA markers. Our aims were to estimate effective population size and the degree of isolation between coastal sites on the mainland of Guinea and Loos Islands. The sampling locations encompassed Dubreka, the area with the highest Human African Trypanosomosis (HAT) prevalence in West Africa, mangrove and savannah sites on the mainland, and two islands, Fotoba and Kassa, within the Loos archipelago. These data are discussed with respect to the feasibility and sustainability of control strategies in those sites currently experiencing, or at risk of, sleeping sickness. PRINCIPAL FINDINGS: We found very low migration rates between sites except between those sampled around the Dubreka area that seems to contain a widely dispersed and panmictic population. In the Kassa island samples, various effective population size estimates all converged on surprisingly small values (10 2 log(10)) by irradiation at 10 kGy or more. However, in the experiment to find out repair of the infectivity, recovery was not noted until day 7 post incubation. PMID- 19290086 TI - A survey of dung beetles infected with larval nematodes with particular note on Copris lunaris beetles as a vector for Gongylonema sp. in Iran. AB - Dung beetles (family Scarabaeidae) are one of the largest families of beetles worldwide. Due to biological behavior of these arthropods, they are considered to play an important role in the life cycle of some helminths. In the present study, dung beetles collected from cattle pastures in rural areas of Ardabil province, north-west of Iran were examined for infection with larval stages of helminths. According to the results, nematodes of 2 genera were identified including Rhabditis and Gongylonema. The more common species was Rhabditis sp. which was found in 9 species of beetles. Out of 15 different species of dung beetles, Copris lunaris was the only scarabaeid to be found naturally infected with the larval stages of Gongylonema sp. Our new findings introduce C. lunaris as a potential biological vector for transmission of Gongylonema sp. to vertebrates in the surveyed region. PMID- 19290087 TI - Changes in sonographic findings after treatment of patients with clonorchiasis in a heavy endemic area. AB - We measured changes in sonographic findings of patients with clonorchiasis after a treatment in a highly endemic area. A total of 347 residents showed positive stool results for Clonorchis sinensis eggs in a village in northeastern China, and were treated with praziquantel. Of them, 132 patients underwent abdominal sonography both before and 1 year after treatment, and the changes in sonographic findings of 83 cured subjects were compared. Diffuse dilatation of intrahepatic bile ducts (DDIHD) was found in 82 patients (98.2%) before and 80 (96.4%) after treatment, which was improved in 3, aggravated in 1, and unchanged in 79 patients. Increased periductal echogenicity (IPDE) was observed in 42 patients (50.6%) before and 45 (54.2%) after treatment, which was improved in 5, aggravated in 8, and unchanged in 70 patients. Floating echogenic foci in the gallbladder (FEFGB) was detected in 32 patients (38.6%) before and 17 (20.5%) after treatment, which was improved in 20, aggravated in 5, and unchanged in 58 patients. Improvement of FEFGB only was statistically significantly (P = 0.004). The present results confirm that DDIHD and IPDE persist but FEFGB decreases significantly at 1 year after treatment. In a heavy endemic area, the sonographic finding of FEFGB may suggest active clonorchiasis 1 year after treatment. PMID- 19290088 TI - Indochinamon ou (Crustacea: Potamidae) as a new second intermediate host for Paragonimus harinasutai in Luang Prabang Province, Lao PDR. AB - Paragonimus harinasutai metacercariae were found in a species of freshwater crab, Indochinamon ou, collected in a small stream of Namback District, Luang Prabang Province, Lao PDR. Adult flukes were recovered after experimental infection of the metacercariae to dogs. Metacercariae were round or slightly elliptical, 0.666 x 0.626 mm in average size, and had a thin cyst wall of about 20 microm in thickness, a black excretory bladder, convoluted ceca, and some pinkish materials in the body. Adults were somewhat elongated, 95.2 x 36.5 mm in average size, covered with single-tipped tegumental spines, had a smaller oral sucker than the ventral sucker, a moderately branched ovary, and 5-6 lobulated testes. Eggs were ovoid and bilaterally symmetrical in shape, 79 x 45 microm in average size, and had a uniformly thickened shell. By the present study, it has been confirmed that I. ou is a new second intermediate host for P. harinasutai. PMID- 19290089 TI - IgE binding reactivity of peptide fragments of Bla g 4, a major German cockroach allergen. AB - Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to full-length and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118 152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy. PMID- 19290090 TI - A nationwide survey on the prevalence of intestinal parasitic infections in the Republic of Korea, 2004. AB - National surveys on the prevalence of intestinal parasitic infections have been carried out every 5-7 years since 1971 in the Republic of Korea in order to establish control measures. The present nationwide survey was conducted from June to December 2004. The 10% population sampling data of Population and Housing Census by the Korean government in 2000 was used as the survey population. One sample was selected randomly from each of the 22,858 registered subjects, and a total of 20,541 people were ultimately included in this survey. Fecal examinations were performed by the cellophane thick smear and saturated brine flotation techniques. Pinworm infection was examined by cello-tape anal swab method. This survey also included a questionnaire study for a socioeconomic analysis. The total helminth egg positive rate was 3.7%, and the estimated total positive number among nationwide people was 1,780,000. The rates in urban and rural areas were 3.1% and 6.8%, respectively. As the total egg positive rate in the 6th survey in 1997 was 2.4%, the present survey showed that there was a considerable degree of increase in the prevalence rate of intestinal parasitic infections over the 7-year period following the 6th survey. The largest increases occurred in the egg positive rates of Clonorchis sinensis and heterophyids including Metagonimus yokogawai. PMID- 19290091 TI - Disseminated microfilaremia associated with lung cyst and empyema: an autopsy report. AB - Clinical manifestations of extralymphatic disease caused by filariasis are varied and range from symptoms due to tropical pulmonary eosinophilia to hematuria, proteinuria, splenomegaly, and rarely arthritis. Disseminated microfilaremia in association with loculated lung cyst and empyema is of rare occurrence and to the best of our knowledge has not been documented in the literature so far. We report here a case of disseminated microfilaremia due to Wuchereria bancrofti infection accompanied by a lung cyst and empyema in a 21-year-old Indian man. PMID- 19290092 TI - Cerebellar schistosomiasis: a case report with clinical analysis. AB - The authors report here a rare case of cerebellar schistosomiasis identified by pathological diagnosis, lacking extracranial involvement. The clinical symptoms included headache, dizziness, and nausea. Studies in blood were normal and no parasite eggs were detected in stool. Computed tomography of brains showed hypodense signal, and magnetic resonance imaging showed isointense signal on T1 weighted images, hyperintense signal on T2-weighted images, and intensely enhancing nodules in the right cerebellum after intravenous administration of gadolinium. A high-grade glioma was suspected, and an operation was performed. The pathologic examination of the biopsy specimen revealed schistosomal granulomas scattered within the parenchyma of the cerebellum. The definitive diagnosis was cerebellar schistosomiasis japonica. A standard use of praziquantel and corticosteroid drugs was applied, and the prognosis was good. When the pattern of imaging examinations is present as mentioned above, a diagnosis of brain schistosomiasis should be considered. PMID- 19290093 TI - External ophthalmomyiasis caused by Oestrus ovis: a rare case report from India. AB - Myiasis of different organs has been reported off and on from various regions in the world. We report a human case of external ophthalmomyiasis caused by the larvae of a sheep nasal botfly, Oestrus ovis, for the first time from Meerut city in Western Uttar Pradesh, India. A 25-year-old farmer presented with severe symptoms of conjunctivitis. The larvae, 3 in number, were observed in the bulbar conjunctiva, and following removal the symptoms of eye inflammation improved within a few hours. PMID- 19290094 TI - A serological study on Toxoplasma gondii infection among people in south of Tehran, Iran. AB - Although Toxoplasma gondii infection generally shows subclinical courses in adults, it may have severe pathologic effects on the fetus. For a survey of toxoplasmosis in Shahr-e-Rey area, south of Tehran, Iran, we conducted a serological study on 1,187 general populations using indirect fluorescent antibody (IFA) test. In this cross sectional study, cases were selected randomly by a local primary health care (PHC) system. Blood samples were collected in microcapillary tubes and sera were examined by IFA after centrifugation. It has been shown that 68.3% of cases in all age groups had IFA titer of 1 : 10 and higher, which we consider as positive cases, and the rate for 1 : 400 or over was 30.2%. There was meaningful correlation between anti-Toxoplasma antibody titers and age, sex, and occupation of subjected people (P < 0.05%). The incidence was higher in females and highest in the children age group (10-14 years old). The present study demonstrates high prevalence of Toxoplasma infection among the people in south of Tehran, Iran. PMID- 19290095 TI - Ultrastructural localization of Toxocara canis larval antigen reacted with a seropositive human serum. AB - Excretory-secretory products of Toxocara canis larvae have been considered as a major functional antigen in immune responses against toxocariasis. We studied ultrastructural localization of T. canis second-stage larval antigen using a seropositive human serum under immunogold electron microscopy. High-density gold particles were observed in the secretory cells, excretory duct, intestinal epithelium, and cuticle of the larval worm sections. The distribution of the positive reactions in the larval worms suggests that the nature of the antigen is excretory-secretory antigen including waste metabolites and secretory enzymes. PMID- 19290096 TI - Separation of the syncytial layer of spargana using urea. AB - The tegument of tapeworms is known to be composed of an outer syncytial cytoplasm layer which includes microtriches and cytoplasmic organelles (= syncytial layer), and a parenchymatous cytoplasm layer that contains subtegumental cell nuclei (= subtegumental layer) and organelles. In the present study, separation of the syncytial layer of the sparganum, the plerocercoid stage of Spirometra mansoni, was tried using urea as the chemical reagent. Histological sections were prepared to visualize the status of separation after staining with hematoxylin and eosin. The results showed that the syncytial layer of the sparganum tegument which includes microtriches and cytoplasmic organelles were successfully separated from the parenchyma using 3 M urea. PMID- 19290097 TI - Immunoblot patterns of Taenia asiatica taeniasis. AB - Differential diagnosis of Taenia asiatica infection from other human taeniases by serology has been tested. An enzyme-linked immunoelectrotransfer blot (EITB) was applied to subjected human sera and tapeworm materials. Thirty-eight proteins reactive to serum IgG were observed between 121 and 10 kDa in adult worms, and more than 22 serum-reactive components between 97 kDa and 21.5 kDa were observed in eggs of T. asiatica. Antigens of adult T. asiatica revealed immunoblot bands between 120 and 21.5 kDa against T. asiatica infected sera. Antigens of adult Taenia saginata revealed 110-100, 66, 58-56, and 46 kDa immunoblot bands against T. asiatica infected sera. Antigens of adult Taenia solium also revealed 99-97, 68-66, and 46 kDa bands against T. asiatica infected sera. The immunoblot band of 21.5 kDa exhibited specificity to T. asiatica. PMID- 19290099 TI - Cytogenetic Status of Meloidogyne (Hypsoperine) spartinae in Relation to Other Meloidogyne Species. AB - Four populations of Meloidogyne spartinae from the coast of North and South Carolina were identical cytogenetically. Fourteen rod-shaped chromosomes were present in oogonia and spermatogonia, whereas seven bivalents were observed in oocytes and spermatocytes. There were no distinguishable sex chromosomes. Chromosome behavior was similar to that of other Meloidogyne species. A slight deviation in morphology of prometaphase bivalents was attributed to an increase in frequency of chiasmata that may be associated with the obligatorily amphimictic reproduction of this nematode. The anatomy of the oviduct spermatotheca region and most cytogenetic features studied suggested that M. spartinae can be regarded as a root-knot nematode. Its position in the genus Meloidogyne or Hypsoperine can be decided by taxonomists. Its small chromosome number (n = 7) compared to the larger number (n = 13-19) of other Meloidogyne species suggests that, cytologically, M. spartinae stands closer to the ancestral form from which the prescent day root-knot nematodes have evolved. PMID- 19290100 TI - Enzymatic relationships and evolution in the genus meloidogyne (nematoda: tylenchida). AB - Thirty populations of Meloidogyne of diverse geographic origin representing 10 nominal species and various reproductive, cytological, and physiological forms known to exist in the genus were examined to determine their enzymatic relationships. The 184 bands resolved in the study of 27 enzymes were considered as independent characters. Pair-wise comparisons of populations were performed in all possible combinations to estimate the enzymatic distances (ED) and coefficients of similarity (S). A phylogenetic tree was constructed. The apomictic species M. arenaria, M. microcephala, M. javanica, and M. incognita shared a common lineage. M. arenaria was highly polytypic, whereas conspecific populations of M. javanica and M. incognita were largely monomorphic. The mitotic and meiotic forms of M. hapla were very similar (S = 0.93), suggesting that the apomictic race B evolved only recently from the meiotic race A. The five remaining meiotic species (M. chitwoodi, M. graminicola, M. graminis, M. microtyla, and M. naasi - each represented by a single population) were not closely related to each other or to the mitotic species. PMID- 19290101 TI - Relationship Between Morphology and Parasitism in Two Populations of Meloidogyne incognita. AB - The reliability of morphological characters and host differential plants for distinguishing between two populations of Meloidogyne incognita was studied. Population A (originally from North Carolina) had incognita-type perineal patterns. A single egg mass subpopulation of population A had a mixture of incognita and acrita perineal patterns with 33% of the patterns atypical for either species. Population B (from Georgia) had predominantly acrita-type patterns with only about 5% atypical patterns. The head shapes of males from both populations were mainly M. incognita. On the basis of stylet length, both populations conformed to M. incognita acrita. Both populations were identified as M. incognita race 1 by reaction on the North Carolina differential hosts. Reactions on azalea and pepper gave no clear identification of the populations. We concluded that there is no relation between perineal pattern, male head shape, and parasitism of host differentials with the two populations studied. PMID- 19290102 TI - Nicotine Content of Tobacco Roots and Toxicity to Meloidogyne incognita. AB - The motility of Meloidogyne incognita second-stage juveniles (J2) and their ability to induce root galls in tomato were progressively decreased upon exposure to nicotine at concentrations of 1-100 mug/ml. EC values ranged from 14.5 to 22.3 mug/ml, but J2 motility and root-gall induction were not eliminated at 100 mug/ml nicotine. Nicotine in both resistant NC 89 and susceptible NC 2326 tobacco roots was increased significantly 4 days after exposure to M. incognita. The increase was greater in resistant than in susceptible tobacco. Root nicotine concentrations were estimated to be 661.1-979.1 mug/g fresh weight. More M. incognita were detected in roots of susceptible than in roots of resistant tobacco. Numbers of nematodes within resistant roots decreased as duration of exposure to M. incognita was increased from 4 to 16 days. Concentrations of nicotine were apparently sufficient to affect M. incognita in both susceptible and resistant tobacco roots. Localization of nicotine at infection sites must be determined to ascertain its association with resistance. PMID- 19290103 TI - Evaluation of Cultivars, Experimental Lines and Plant Introduction Collection of Sainfoin for Resistance to Meloidogyne hapla Chitwood. AB - Stands of several cultivars and experimental lines of sainfoin (Onobrychis viciifolia) were severely reduced (92% average loss) in a field naturally infested with Meloidogyne hapla. Stands of two alfalfa cultivars included in the test were unaffected. In studies conducted in the greenhouse with plants inoculated at the time of seeding, average mortality was 55% for sainfoin entries and 7% for Ladak alfalfa. Little mortality occurred when plants were inoculated after establishment. Three months after inoculation, all sainfoin entries were heavily galled (range of 3.3-3.7 on a scale of 1-4) while roots of Ladak were only slightly galled (rating of 1.6). Intermating of plants selected in the field plots for resistance to M. hapla showed a slight increase in resistance. Of the 147 plant introduction lines tested in the greenhouse, none were resistant to M. hapla. PMID- 19290104 TI - Root-knot Nematode Management and Yield of Soybean as Affected by Winter Cover Crops, Tillage Systems, and Nematicides. AB - Management of Meloidogyne incognita on soybean as affected by winter small grain crops or fallow, two tillage systems, and nematicides was studied. Numbers of M. incognita did not differ in plots planted to wheat and rye. Yields of soybean planted after these crops also did not differ. Numbers of M. incognita were greater in fallow than in rye plots, but soybean yield was not affected by the two treatments. Soybean yields were greater in subsoil-plant than in moldboard plowed plots. Ethylene dibromide reduced nematode population densities more consistently than aldicarb and phenamiphos. Also, ethylene dibromide increased yields the most and phenamiphos the least. There was a positive correlation (P = 0.001) of seed size (weight of 100 seeds) with yield (r = 0.79), indicating that factors affecting yield also affected seed size. PMID- 19290105 TI - Relative susceptibility of four pine species to infection by pinewood nematode. AB - Mature trees of eastern white, jack, Scotch, and shortleaf pines were inoculated with 25,000-34,000 pinewood nematodes, Bursaphelenchus xylophilus, isolated from infected Scotch pines in Missouri. Equal numbers of trees of each species inoculated with distilled water served as controls. Nine of fifteen Scotch pines died within 4 months of nematode infection or during the winter and early spring following infection. A single eastern white and shortleaf pine died. No jack pines died. A single Scotch pine control died, apparently the result of natural nematode infection. No other controls died. Mean oleoresin flow did not differ among nematode-inoculated jack and shortleaf pines and their respective controls. Oleoresin flow in nematode-inoculated eastern white and Scotch pines was significantly lower than in their controls. Oleoresin flow was temporarily reduced in mortality-resistant eastern white and Scotch pines following nematode infection. Thus a sublethal impact of nematode infection on mortality-resistant host trees was documented. PMID- 19290106 TI - Carbohydrate Concentration in Pine as Affected by Inoculation with Bursaphelenchus xylophilus. AB - Pines responded to inoculation with Bursaphelenchus xylophilus by changes in reducing and nonreducing carbohydrate concentrations dependent on the pine species and the pathotype of B. xylophilus with which the trees were inoculated. Carbohydrate concentrations, in compatible pine-nematode pathotype combinations, decreased initially after inoculation and then increased slightly before decreasing to approximately 10% of the control levels as the seedlings wilted. In compatible nematode pathotype-pine species combinations, carbohydrate concentrations decreased and then increased as the nematode population densities declined. PMID- 19290107 TI - Descriptions of Two New Species of Tylenchorhynchus Cobb, 1913 (Nematoda: Tylenchida), with Details on Morphology and Variation of T. claytoni. AB - Two new species of plant parasitic nematodes (Tylenchorhynchus quaidi n. sp. and T. tritici n. sp.) from Pakistan are described and illustrated. Tylenchorhynchus quaidi n. sp., from soil around roots of potato (Solanum tuberosum) from an experimental field of NNRC, Karachi, Pakistan, is distinguishable from other species by its peculiar sunken dome-shaped head. Although similar to T. goffarti, it differs by head shape, areolation of lateral field, ratios a (23-28 vs. 29-37) and c (11-14 vs. 13-20), and a vagina that is half sclerotized and half unsclerotized. Tylenchorhynchus tritici n. sp., from soil around roots of wheat (Triticum aestivum) from Campbellpur, Pakistan, is similar to T. ventrosignatus and T. nordiensis. It differs from T. ventrosignatus by a continuous lip region, number of head annules (2-3 vs. 4), coarse body annulation, absence of a wave like structure near the vulva, and by tail shape and number of tail annules (15 23 vs. 28-32). It differs from T. nordiensis by stylet length (12.4-14.6 vs. 11 13 mum), shape of stylet knobs, number of head annules (2-3 vs. 4), non-areolated lateral field in region of phasmids, and not fusing in posterior third of tail. Morphometrics of Tylenchorhynchus claytoni from soil around stunted maize (Zea mays L.), in Muscatine County, Iowa, and several other populations are given. Detailed morphometric data on T. claytoni based on topotypes collected from type locality and several other populations revealed that this species shows variations in the shape of tail in females, number of tail annules (and sometimes annules extending further back on the terminus, almost being an annulated terminus), position of phasmid, and shape of lip region. The subgenus Bitylenchus is proposed as a new synonym of Tylenchorhynchus and its species referred to the latter genus. PMID- 19290108 TI - New records of paratylenchus micoletzky, 1922 from Brazil with descriptions of two new species (tylenchulidae: nemata). AB - Six species of Paratylenchus Micoletzky, 1922, two of them new, are reported from the Amazon Basin of Brazil. P. colbrani is reported for the first time in Brazil. Extended ranges, along with additional measurements and descriptions, are given for P. leptus, P. salubris, and P. perlatus. P. mimulus Raski, 1975 is synonymized with P. salubris Raski, 1975. P. flectospiculus n. sp. is distinguished by four lines in the lateral field, short stylet (21-24 mum), and spicules with distinct distal bend ventrad. P. rostrocaudatus n. sp. is distinguished by three lines in the lateral field, short stylet (15-16 mum), and beaked tail terminus. PMID- 19290109 TI - Revised Host Range and Studies on the Life Cycle of Longidorus africanus. AB - The host range of Longidorus africanus was demonstrated to be much wider than previously reported. All commercial crop plants tested, except two of four crucifers, were hosts of L. africanus. The nematode was widespread in fields, and soil type did not appear to be related to its distribution. The minimum time to complete a life cycle was 9 weeks at 28 C in a plant growth chamber. Field observations of population densities indicated, however, that in undisturbed field soils the life cycle required considerably less time than was indicated by growth chamber studies. PMID- 19290110 TI - Hirschmanniella spp. in Rice Fields of Vietnam. AB - Root and soil samples from one-crop and two-crop rice fields were collected in a survey for Hirschmanniella spp. in Vietnam during 1978-80. Hirschmanniella spp. were found in 50-78% of the soil samples and 98-100% of the root samples collected. Population densities of nematodes in root systems were lowest at posttransplanting and highest at heading time. Numbers of nematodes inside roots increased 20-22 times from transplanting to heading in fields with both crop sequences. Population densities of Hirschmanniella spp. in two-crop rice fields were more than twice those in one-crop rice fields. PMID- 19290111 TI - Influence of Edaphic Factors and Previous Crop on Pratylenchus spp. Population Densities in Potato. AB - Root and soil samples from commercial potato fields were assayed for nematodes in 1983 and 1984. Pratylenchus spp. population densities in Suffolk County, New York, were consistently, though not always statistically, higher in potato fields that had been planted to rye or wheat rather than potatoes during the previous growing season. Regardless of the previous crop, population densities in the two potato production areas in Suffolk County differed significantly: population densities on the south fork were 1.9-5.5 times higher than those on the north fork. Species prevalence differed significantly on the two forks but was not related to the previous year's crop. P. penetrans and P. crenatus were found primarily on the north and south forks, respectively. Differences in species distribution were associated with differences in soil types. P. crenatus was usually found on loams and silt loams, but P. penetrans was found more frequently on sandy soils. PMID- 19290112 TI - Effects of Soil Solarization on Rotylenchulus reniformis in the Lower Rio Grande Valley of Texas. AB - Soil solarization was evaluated for control of Rotylenchulus reniformis in the lower Rio Grande Valley of Texas. In field experiments, solarization significantly reduced soil nematode population densities 0-15 cm deep and increased yields of lettuce and cowpea. The length of time required for 90% mortality of nematodes in soil heated under controlled conditions in the laboratory varied from 25 hours to less than 1 hour between 41 and 47 C. Daily exposures of nematode-infested soil to lethal temperatures for sublethal time periods had a cumulative lethal effect. In water, vermiform stages required up to 10 days to recover from sublethal thermal stress. Eggs were similar to juveniles in their sensitivity to high temperatures. Lethal time-temperatures under controlled conditions were in general agreement with field results. PMID- 19290113 TI - Extraction of cyst nematodes from organic soils. AB - The effects of extraction technique, sample size, soil moisture level, and overflow rate on recovery of Globodera rostochiensis and (or) Heterodera schachtii cysts from organic soils were investigated. A modified Fenwick can (MFC) and an underflow elutriator (UE) described in this paper were evaluated and compared for cyst recovery efficiency and amount of organic flotsam collected. The MFC and UE extracted similar numbers of cysts, but the UE collected 50% less flotsam than the MFC. Sample size was negatively correlated with cyst recovery and positively correlated with amount of flotsam. The amount of flotsam recovered with the MFC was correlated with overflow speed. Presoaking air dried samples for 30 minutes halved the amount of flotsam without affecting cyst recovery. Extracting cysts from wet soil without prior drying resulted in negligible recovery with both extraction techniques. There were no significant differences in cyst recovery of the two genera tested. PMID- 19290114 TI - Nematode-degree days, a density-time model for relating epidemiology and crop losses in perennials. AB - The impact of Meloidogyne hapla on alfalfa (Medicago sativa) yield was described by a multiple point damage model as a function of current plant status, cumulative pest stress, and crop history. Nematode-degree days (NDD(female symbol)), calculated on a physiologic time scale as total area under the adult female population density curve, were used to express M. hapla parasitism as cumulative nematode dosage. NDD(female symbol) increased exponentially over physiological time at rates relative to M. hapla initial population density of eggs and second-stage juveniles (Pi). Dosage accumulation rates, varying from 213 to 272 NDD(female symbol) per degree day, did not differ (P = 0.05) among six Pi levels. Cumulative yield reductions, increasing linearly with NDD(female symbol) over time to 50% of the M. hapla-free controls, were well correlated for the six initial Pi levels (r(2) = 0.93). Progressive reductions in alfalfa yields to 65% of the nematode-free controls, reflective of the prolonged exposure of the crop to M. hapla, were adequately described by NDD model estimates of either combined population densities of eggs and juveniles or adult females. Cumulative area under the combined eggs (e) and juvenile (J) population curve (NDD(e+J)) and NDD(female symbol) were linearly related (r(2) = 0.97). PMID- 19290115 TI - Movement of potato root diffusate through soil. AB - Movement of potato root diffusate (PRD) through soil was examined by using the hatch of eggs from Globodera rostochiensis cysts as an indicator. Porous bags containing cysts were placed at increasing distances and depths from potato roots, whose growth was restricted by nylon mesh. Significantly greater hatch was observed up to 50 cm laterally away from potato roots, compared with hatch in fallow soil. Eight weeks after plant emergence, we detected a concentration gradient of PRD, as measured by egg hatch, that decreased with increasing lateral and vertical distance from the root zone. Egg hatch beyond 5 weeks after plant emergence was not attributed to PRD. PMID- 19290116 TI - In Vivo Growth of Romanomermis culicivorax: Biochemical Changes During Parasitism. AB - Biochemical analyses of total protein, lipid, carbohydrate, DNA, amino acid, and length, width, and dry weight measurements are reported for different stages of Romanomermis culicivorax cultured in the mosquito, Culex pipiens. The Bradford technique for assaying total protein was the most sensitive and reliable biochemical technique tested for assaying in vivo growth of R. culicivorax. Increases in total protein, lipid, carbohydrate, and dry weight during growth from preparasite to postparasite were greater than 6,900-fold for females and 2,300-fold for males. DNA increased 650-fold and 233-fold during development to female and male postparasites, respectively. The proportions of amino acids for preparasites were significantly different (P 7 cm) in the Vorlex + Temik plots was 20.7 t/ha, compared to 4.7 t for the untreated check. Vorlex alone and Telone II B plots yielded 17.3 and 15.9 t/ha, respectively; Temik with 7.5 t also yielded better than the check. Vydate and Furadan did not influence yields significantly. Total yields differed from the check in all treatments except with Furadan. In 1984, marketable yields ranged from 15.5 t/ha for the Vorlex + Temik treatment to 11.2 for the untreated check, but the differences were not statistically significant. Total yields, however, were significantly increased by the fumigants. The difference in response to chemical treatment in the 2 years was attributed to greater heat stress and lack of supplementary irrigation in 1983. PMID- 19290167 TI - Two-dimensional Protein Patterns in Labronema, Aporcelaimellus, and Eudorylaimus (Nematoda: Dorylaimida). AB - Two-dimensional polyacrylamide gel electrophoretic patterns of proteins for two isolates of Labronema from Indiana were nearly identical to the pattern for L. vulvapapillatum from Europe. The pattern for a nominal isolate of L. pacificum from Florida was very different from the patterns of nominal L. pacificum isolates from Hawaii and Fiji (which had patterns very similar to each other). Patterns for four other isolates (in Eudorylaimus and Aporcelaimellus) were different from the Labronema patterns and from each other, although some constellations of protein spots were shared among all the isolates. The study demonstrates the utility of 2-D PAGE for clarifying taxonomic problems that cannot be resolved using classical morphological data alone. PMID- 19290168 TI - Interaction of Ditylenchus dipsaci and Meloidogyne hapla on Resistant and Susceptible Plant Species. AB - Numbers ofDitylenchus dipsaci or Meloidogyne hapla invading Ranger alfalfa, Tender crop bean, Stone Improved tomato, AH-14 sugarbeet, Yellow sweet clover, and Wasatch wheat from single inoculations were not significantly different from numbers by invasion of combined inoculations. D. dipsaci was recovered only from shoot and M. hapla only from root tissue. Combined inoculations did not affect reproduction of either D. dipsaci or M. hapla. D. dipsaci suppressed shoot growth of all species at 15-30 C, and M. hapla suppressed shoot growth of tomato, sugarbeet, and sweet clover at 20, 25, and 30 C. There was a positive correlation (P < 0.05) between shoot and root growth suppression by D. dipsaci on all cultivars except wheat at 20 C and tomato at 30 C. M. hapla suppressed (P < 0.05) root growth of sugarbeet at 20-50 C and wheat at 30 C. Growth suppression was synergistic in combined inoculations of sweet clover shoot growth at 15 C and root growth at 20-30 C, wheat root growth at 15 and 20 C, and tomato root growth at 15-30 C (P < 0.05) D. dipsaci invasions caused mortality of alfalfa and sweet clover at 15-30 C and sugarbeet at 20-30 C. Mortality rates of alfalfa and sweet clover increased synergistically (P < 0.05) from combined inoculations. PMID- 19290169 TI - Effect of Hirschmanniella caudacrena on the Submersed Aquatic Plants Ceratophyllum demersum and Hydrilla verticillata. AB - In vitro pathogenicity tests demonstrated that Hirschmanniella caudacrena is pathogenic to Ceratophyllum demersum (coontail). Symptoms were chlorotic tissue, deformed stems, and, finally, death of the plant. Inoculum densities of 500 nematodes per 5-cm-long cutting in a test tube containing 50 ml of water resulted in death and decay of some of the cuttings within 8 weeks; 100 nematodes killed the plants in 12 weeks, and 50 and 25 nematodes killed them in 16 weeks. The lowest inoculum level of 10 nematodes did not seriously affect the plants at 16 weeks when the experiment was terminated. A second test conducted outdoors in glass jars containing 3 liters of water and two cuttings weighing a total of 15 g fresh weight showed damage, but results were not statistically significant. Hydrilla verticillata inoculated with H. caudacrena was not affected seriously. PMID- 19290170 TI - Control of Pratylenchus penetrans and Meloidogyne hapla and Yield Response of Alfalfa Due to Oxamyl Seed Treatments. AB - Alfalfa (Medicago sativa L. cv. Saranac) seed were soaked for 20 minutes in water, acetone, or methanol containing 10 or 50 mg/ml of oxamyl (Vydate L) or coated with a 2% aqueous cellulose solution containing the same amounts of oxamyl. Seed were analyzed for oxamyl by HPLC immediately after treatment and after 9 and 26 months of storage. Oxamyl content of alfalfa seed did not decline after 26 months of storage. The effects of seed treatment on growth of alfalfa and nematode control were examined using soils infested with Pratylenchus penetrans and Meloidogyne hapla. Germination was not affected by any of the seed treatments. Twenty-one days after sowing, the total growth of alfalfa seedlings grown from seed treated with 50 mg/ml of oxamyl in P. penetrans-infested soils had increased by 62% over controls. Nodulation per pot increased by as much as 267%, and the densities of P. penetrans per gram of root were reduced by as much as 73% compared to control plants. In M. hapla-infested soils, increases in plant growth (32%) and nodulation (71%) also occurred with oxamyl-treated seeds. Root gall reduction (86%) was also substantial due to oxamyl seed treatment. PMID- 19290171 TI - Selection Against Heterodera glycines Males by Soybean Lines with Genes for Resistance. AB - Soybeans with genes for resistance select against Heterodera glycines with the corresponding genes for avirulence. There may be a differential effect of sex with some specific gene interactions, which would influence the magnitude of gene frequency changes. No effect on H. glycines males was detected with one selected nematode population and the resistant soybean line PI88788. The selective effect of PI89772 against male nematodes was greater with two inbred nematode populations than with one selected (on PI88788) population, presumably due to differences in H. glycines gene frequencies. 'Peking' also had few males with the one inbred nematode population, whereas Forrest and 'Pickett 71' had intermediate numbers. Apparently Forrest and Pickett 71 did not get all the Peking genes for resistance that affect male as well as female nematode development. Other H. glycines-soybean genes stop only females, since there were few or no cysts, except on the susceptible soybean Williams. The number of males' phenotype will help identify specific genes in both organisms. PMID- 19290172 TI - Production and Partial Characterization of Stylet Exudate from Adult Females of Meloidogyne incognita. AB - Adult females of Meloidogyne incognita were excised from tomato roots and incubated in 0.04 M phosphate buffered saline, pH 7.4 for 18-72 hours to allow accumulation of stylet exudate. Twenty-four percent of the females produced exudate during the initial 18-hour incubation period; 70% of those females producing exudate initially produced additional exudate during the subsequent 54 hour incubation period. Analysis of exudate by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed the presence of at least nine major protein bands. Differential staining with silver and Coomassie Brilliant Blue G 250 stains indicated that three of the bands were glycoproteins. Upon acid hydrolysis, 14 amino acids were detected in the stylet exudate. The basic amino acids lysine, histidine, and arginine comprised 21.8% of the total amino acids detected. No peroxidase activity was detected in the stylet exudates. Data presented extend and generally confirm prior work on the chemical composition of stylet exudate. PMID- 19290173 TI - Ultrastructure of Esophageal Gland Secretory Granules in Juveniles of Heterodera glycines. AB - Ultrastructural observations of the feeding sites of soybean cyst nematode juveniles 3 days after inoculation of soybean roots revealed the presence of feeding tubes in the host cell syncytium. Feeding tubes, which were extruded from the stylet tips, were formed by products of secretory granules that originated in the dorsal esophageal gland and accumulated in the ampulla of the gland extension. Granules traversing the space between the gland cell and the ampulla were regulated in their movement by two sets of sphincter-like muscles located anterior and posterior to the metacorpus pump chamber. Sections through the sphincter muscles revealed obliquely arranged fibers, which in a contracted mode caused microtubules in the gland extension to be tightly packed and devoid of granules. PMID- 19290174 TI - Peanut-Cotton Rotations for the Management of Meloidogyne arenaria. AB - The efficacy of 'Deltapine 90' cotton (Gossypium hirsutum) in rotation with 'Florunner' peanut (Arachis hypogaea) for the management of Meloidogyne arenaria was studied for 2 years in a field in southeastern Alabama. In 1985, M. arenaria juvenile populations in plots with cotton were 98% lower than in plots with peanut. Peanut and cotton yields were increased by treatment with aldicarb (3.3 kg a.i./ha in a 20-cm-band) in 1985 but not in 1986. In 1986, peanut yields were highest and M. arenaria juvenile populations in soil were lowest in plots that had cotton the previous year. In 1986, numbers of M. arenaria juveniles in plots with peanut both years were reduced by treatment with aldicarb to levels found in plots with cotton-peanut rotation. The use of aldicarb in peanut following cotton similarly treated reduced the incidence of southern blight (Sclerotium rolfsii). Cotton-peanut is a good rotation for the management of M. arenaria and to increase peanut yields without the use of nematicides. PMID- 19290175 TI - Nutrient Composition of Romanomermis culicivorax in Relation to Egg Production and Metabolism. AB - The nutrient composition of postparasitic females (newly emerged juveniles, newly molted adults, and spent adults) and eggs of Romanomermis culicivorax was studied. Throughout post-parasitic development, proteins increased and lipids decreased progressively as a proportion of the dry weight; the proportion of glycogen within the nematodes remained stable. The greatest decrease in the lipid moiety occurred during egg production. Eggs contained relatively low levels of lipids (12% dry weight), and ca. 20% of the dry weight of the eggs was unaccounted for by lipid, protein, and glycogen determinations. Chitin, mucoproteins, and peptides were present in the eggs. The fatty acid composition of nematodes remained constant during postparasitic development; eggs contained a similar profile of fatty acids as postparasites, with marginally higher content of unsaturated fatty acids. Radiotracer studies showed that the eggs could oxidize glucose and palmitic acid. PMID- 19290176 TI - Ecological study of nematode parasitism in ips beetles from california and idaho. AB - Nematodes found in Ips paraconfusus from ponderosa pine in California were an undescribed species of Parasitaphelenchus, Contortylenchus elongatus, C. reversus, and C. brevicomi. C. elongatus, the most commonly found contortylenchid, was present in 98.2% of the contortylenchid-parasitized beetles. Only one nematode parasite of the gut, a Parasitorhabditis sp., was isolated. Although significant differences in parasitism were observed, they were by collection sites, rather than by elevation or bole sources (slash or standing). Significant changes in parasitism between fall and spring collections were observed but not at every site. Nematode parasitism in the F generation of I. paraconfusus by Parasitaphelenchus, Contortylenchus, or Parasitorhabditis increased or decreased from the parent generation depending upon the experiment.Nematode parasites from I. pini included an undescribed Parasitaphelenchus sp., two undescribed Contortylenchus spp., C. reversus and Parasitylenchus (= Neoparasitylenchus) ovarius from the hemocel, and Parasitorhabditis ipini from the gut. Parasitaphelenchus sp. was found in 99% and 45.3% of the beetles from Idaho and California, respectively. Of the 1,000 I. pini from Idaho and California, 157 were parasitized by the contortylenchid species or P. ovarius. PMID- 19290177 TI - Comparative Small-scale Field Application of Steinernema feltiae for Navel Orangeworm Control. PMID- 19290178 TI - Embryonic and Postembryonic Development of Meloidogyne californiensis Abdel Rahman &Maggenti, 1987. PMID- 19290179 TI - DNA Isolation and GC Base Composition of Four Root-knot Nematode (Meloidogyne spp.) Genomes. AB - Phenol extraction and cesium trifluoroacetate ultracentrifugation were compared for efficiency in the extraction of DNA from eggs and second-stage juveniles of four species of Meloidogyne. The second method proved to be more satisfactory in that it yielded larger amounts of DNA, shortened the extraction period, and reduced sample handling by eliminating phenol and ether extraction and RNAse treatment. It also made possible the extraction of DNA: from more than one sample at a time. The mean base compositions (% GC) of the total DNA of M. incognita, M. javanica, M. arenaria, and M. hapla, as determined by thermal denaturation tests, were quite similar, as they ranged only between 31 and 33%. Similarly, the thermal stability of the DNA of all four species covered a narrow range from 82.97 to 83.63 C. PMID- 19290180 TI - Three new species of etamphidelus andrassy, 1977 (nemata: alaimidae) in southern chile. AB - Three new species of Etamphidelus are described from Orange Bay, Hoste Island, Chile. All three are distinguished from previously described species by their numerous longitudinal cuticular ridges. E. acucephalus n. sp. is further distinguished by its extremely narrowed anterior body region and posteriorly situated amphids. E. fueguensis n. sp. is distinguished from E. acucephalus by its anteriorly located amphideal fovea, fewer cuticular ridges, smaller V-an/tail ratio and presence of males. E. yamani n. sp. is more similar to E. fueguensis n. sp. differing from it by a wider head end, more posteriorly located excretory pore, longer V-an/tail ratio, more numerous cuticular ridges and smaller spermatozoa. E. puccinelliae (Lorenzen, 1966) Andrassy, 1977 is transferred to Paramphidelus puccinelliae (Lorenzen, 1966) n. comb. The generic diagnosis of Etamphidelus is amended, and a key to species is presented. PMID- 19290181 TI - Histopathology of Root-knot Nematode (Meloidogyne incognita) Infection on White Yam (Dioscorea rotundata) Tubers. AB - White yam tissues naturally and artificially infected with root-knot nematodes were fixed, sectioned, and examined with a microscope. Infective second-stage juveniles of Meloidogyne incognita penetrated and moved intercellularly within the tuber. Feeding sites were always in the ground tissue layer where the vascular tissues are distributed in the tubers. Giant cells were always associated with xylem tissue. They were thin walled with dense cytoplasm and multinucleated. The nuclei of the giant cells were only half the size of those found in roots of infected tomato plants. Normal nematode growth and development followed giant cell formation. Females deposited eggs into a gelatinous egg mass within the tuber, and a necrotic ring formed around the female after eggs had been produced. Second-stage juveniles hatched, migrated, and re-infected other areas of the tuber. No males were observed from the tuber. PMID- 19290182 TI - Detection of Intraspecific Diversity of Heterodera glycines Using Isozyme Phenotypes. AB - Twelve populations of Heterodera glycines from the United States (8), China (2), Japan (1), and Colombia (1) were surveyed for phenotypic intraspecific variability in 42 enzyme systems. Activity of 20 enzymes was detected following isoelectric focusing in polyacrylamide gels of extracts from mass homogenates and single females. Five enzymes, aspartate aminotransferase, phosphoglucose isomerase, alpha- and beta-esterases, and hexokinase were the most useful for detecting intraspecific variability. Phenotypic variability between single females was best demonstrated with alpha- and beta-esterases and acid phosphatase enzyme systems. These results suggest that isoelectric focusing in conjunction with sensitive enzyme systems can be used to detect phenotypic variation between individual nematodes from the same population. The unusual phenotypic variability detected in the H. glycines population from Virginia indicates that the genetic diversity of this population is complex. PMID- 19290183 TI - Behavior, Parasitism, Morphology, and Biochemistry of Criconemella xenoplax and C. ornata on Peach. AB - Host-parasite relationships of Criconemella xenoplax and C. ornata on Nemaguard peach and common bermudagrass were determined in the greenhouse. Criconemella xenoplax reproduced on peach and reduced root volume, height, and dry stem weight after 6 months, compared with the noninfested check. Numbers of C. ornata did not increase on peach or influence peach growth, but they did reduce dry top weight and root volume of common bermudagrass, compared with C. xenoplax. Criconemella xenoplax and C. ornata produced the enzyme beta-glucosidase and were capable of metabolizing prunasin, but only C. xenoplax produced beta-cyanoalanine synthase to detoxify the cyanide released from prunasin. The apparent inability of C. ornata to detoxify cyanide is one explanation why numbers of this species did not increase on peach. Criconemella xenoplax and C. ornata can be distinguished by using stylet length, vaginal configuration, and shape of the anterior head region. PMID- 19290184 TI - Morphological Variation in Xiphinema spp. from New York Orchards. AB - Xiphinema specimens were collected from orchards in southeastern, northeastern, and western New York. Total length, distance of vulva from anterior end, spear length (odontostyle plus odontophore), body diameter at vulva, tail length, anal body diameter, and length and diameter of hyaline tail tip were measured on fixed, glycerol-infiltrated adult females. Most specimens were identified as X. americanum or X. rivesi, but one western New York population was identified as X. californicum (a new record for New York). Multivariate analyses indicated that, with one exception, western New York populations of both X. americanum and X. rivesi were smaller and slimmer than their eastern counterparts. Regional differences were generally larger than differences attributed to host species. PMID- 19290185 TI - Meloidogyne mayaguensis n. sp. (Meloidogynidae), a Root-knot Nematode from Puerto Rico. AB - Meloidogyne mayaguensis n. sp. is described and illustrated from specimens obtained from galled roots of eggplant, Solanum melongena L., from Puerto Rico. The perineal pattern of females is round to ovoid with fine, widely spaced striae. It has occasional breaks of striation laterally and a circular tail tip area lacking striae. The stylet, 15.8 mum long, has reniform knobs that merge gradually with the stylet shaft. Males have a high, rectangular, smooth head region, not set off from the body contour. The labial disc is continuous with the medial lips which do not slope posteriorly. The styler, 22.9 mum long, has large rounded backward sloping knobs; the shaft is of uneven diameter. Mean body length of second-stage juveniles is 453.6 mum. The truncate head region is not annulated, and the rounded, slightly raised labial disc and the crescentic medial lips form dumbbell-shaped lip structures. The stylet, 11.6 mum long, has rounded, posteriorly sloping knobs. The slender tail, 54.4 mum long, gradually tapers to a bluntly pointed tip. Tomato, tobacco, pepper, and watermelon are good hosts; cotton and peanut are not hosts. M. mayaguensis n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of 2n = 44-45. The enzyme patterns are unique among Meloidogyne species. PMID- 19290186 TI - A method for staining nematode secretions and structures. AB - Secretions from amphids, phasmids, and excretory system were stained by incubating nematodes in 0.1% coomassie brilliant blue G-250 in 40% aqueous methanol containing 10% acetic acid on slides with coverslips sealed with nail polish or Zut. Nematodes incubated in this staining solution usually produced copious amounts of secretions from their amphids and excretory pore. Phasmids also stained dark blue, enabling them to be easily observed. Other biological dyes stained these secretions or were useful for differentiating specific morphological features of nematodes. PMID- 19290187 TI - Interactions Between Meloidogyne incognita and Pratylenchus brachyurus on Soybean. AB - Interactions among Meloidogyne incognita, Pratylenchus brachyurus, and soybean genotype on plant growth and nematode reproduction were studied in a greenhouse. Coker 317 (susceptible to both nematodes) and Gordon (resistant to M. incognita, susceptible to P. brachyurus) were inoculated with increasing initial population densities (Pi) of both nematodes individually and combined. M. incognita and P. brachyurus individually usually suppressed shoot growth of both cultivars, but only root growth on Coker 317 was influenced by a M. incognita x P. brachyurus interaction. Reproduction of both nematodes, although dependent on Pi, was mutually suppressed on Coker 317. P. brachyurus reproduced better on Gordon than on Coker 317 but did not affect resistance to M. incognita. Root systems of Coker 317 were split and inoculated with M. incognita or P. brachyurus or both to determine the nature of the interaction. M. incognita suppressed reproduction of P. brachyurus either when coinhabiting a half-root system or infecting opposing half-root systems; however, P. brachyurus affected M. incognita only if both nematodes infected the same half-root system. PMID- 19290188 TI - Trichodorus petrusalberti n. sp. (Nematoda: Trichodoridae) from Rice with Additional Notes on the Morphology of T. sanniae and T. rinae. AB - A new species in the family Trichodoridae, from the rhizosphere of rice (Oryza sativa L.) in Northern Natal, South Africa, is described and illustrated. Trichodorus petrusalberti n. sp. resembles T. taylori De Waele, Mancini, Roca, and Lamberti, 1982, T. hooperi Loof, 1973, and T. complexus Rahman, Jairajpuri, and Ahmad, 1985, from which it is distinguished by the shape of the spicules. It is distinguished from the former two species by the nonthickened terminal cuticle in the males and by the shape and sclerotization of the vulval-vaginal region in lateral view in the females. Additional morphological details are given for T. sanniae Vermeulen and Heyns, 1985 and T. rinae Vermeulen and Heyns, 1985. PMID- 19290189 TI - Effect of Host Age and Nematode Strain on Susceptibility of Spodoptera frugiperda to Steinernema feltiae. AB - Median lethal concentrations (LC) were determined for four nematode populations (two strains of Steinernema feltiae, a S. feltiae hybrid, and S. bibionis) against fifth-instar fall armyworm (Spodoptera frugiperda) larvae and for the most virulent of these nematodes against different instars and stages of the insect. Based on lack of overlap of 95% fiducial limits, there were significant differences in virulence among the four nematodes. The LC ranged from 7.6 to 33.3 nematodes/ 0.7 ml water, and slopes of the log dose-probit regression lines were similar except for the S. feltiae All strain. First-instar fall armyworms suffered virtually 100% mortality from the S. feltiae Mexican strain at 1.0 nematode/0.7 ml, and LC were 2.3 and 7.9 nematodes/0.7 ml in third-instar and fifth-instar larvae, respectively. Pupae had 7-20% mortality at doses ranging from 30 to 60 nematodes/0.7 ml. PMID- 19290190 TI - Parasitism of Kenyan Mosquito Larvae (Diptera: Culicidae) by Romanomermis culicivorax (Nematoda: Mermithidae). AB - The ability of Romanomermis culicivorax to infect, develop, and emerge from Kenyan mosquito hosts was evaluated in the laboratory. Host species tested were Aedes aegypti, Ae. dentatus, Ae. hirsutus, Anopheles arabiensis, An. coustani, An. funestus, An. gambiae, An. pharoensis, Culex duttoni, Cu. ethiopicus, Cu. poicilipes, Cu. quinquefasciatus, Cu. tigripes, Cu. univittatus, Coquillettidia metallica, Mansonia africana, Ma. uniformis, Mimomyia splendens, Mi. uniformis, Toxorhyncites brevipalpis, and Uranotaenia balfouri. R. culicivorax penetrated all the host species tested and developed and emerged from most of the hosts. Both penetration and some development, but not nematode emergence, were observed from all instars of Ma. uniformis. T. brevipalpis exhibited signs of resistance in the form of melanization of R. culicivorax within 48 hours of infection in all four instar stages. Nematode melanization, especially in older hosts, was observed in Ae. dentatus, Ae. hirsutus, Cu. duttoni, Cu. tigripes, and Mi. splendens. When melanization occurred, the melanized carcass of the nematode was passed on from instar to instar. The implications for field release of R. culicivorax in Kenya are still good, especially in habitats where different mosquito species occupy the same niche at different times, which would allow for nematode recycling. PMID- 19290191 TI - Phylogenetic analyses in dorylaimida using data from 2-d protein patterns. AB - Data from two-dimensional protein patterns for nine dorylaimid isolates were analyzed using PAUP, a computer program for inferring phylogenies under the principle of maximum parsimony. With a variety of available options, including branch swapping and rooting, essentially the same tree was obtained. When isolates of the genus Labronema were analyzed alone, all trees obtained had the same topology, although tree length varied considerably, depending on whether a hypothetical ancestral taxon was included. PMID- 19290192 TI - Effects of Host Resistance on the Fecundity of Globodera rostochiensis. AB - The fecundity of Globodera rostochiensis (RA) females that developed on resistant Rosa and susceptible Katahdin potato cultivars were compared. Cysts collected from each cultivar were bulked, separated into four sizes (> 500 mum, 355-500 mum, 250-355 mum, and < 250 mum), and crushed to determine fecundity as measured by viable egg content (VEC). Fewer and generally smaller cysts developed on Rosa than on Katahdin. Although cyst size significantly (P = 0.01) influenced VEC, cyst age (8 or 13 weeks) had no effect. Regardless of size, cysts produced on Rosa contained significantly fewer viable eggs than did cysts produced on Katahdin. The fecundity of progeny from cysts produced on Rosa was significantly reduced compared with that of progeny from cysts produced on Katahdin. After two generations on Katahdin, the VEC of cysts from a population originating from Rosa was significantly less than that of cysts from a population originating from Katahdin, indicating that in the presence of a pure population of G. rostochiensis RA, the females that develop on the resistant cultivar Rosa represent a diminished rather than a superior selected population. PMID- 19290193 TI - Winter Survival of Meloidogyne incognita in Six Soil Types. AB - Winter survival of Meloidogyne incognita in six soil types (Fuquay sand, Norfolk loamy sand, Portsmouth loamy sand, muck, Cecil sandy clay loam, and Cecil sandy clay) was determined in microplots at one location from November 1981 to May 1982 and from November 1982 to March 1983. Survival, based on second-stage juveniles (J2) of M. incognita, from November 1981 until May 1982 ranged from 1% in the muck soil to 6% in a Cecil sandy clay loam, but survival rates were much higher the next year following a winter with higher average temperatures. Survival rates of J2 from November to March ranged from 20 to 40% the first winter and from 38 to 87% the second. Soil type did not have a striking effect on the overwintering capabilities ofM. incognita. There were no differences between clay and sand soils, whereas survival of J2 in the muck tended to be lower than in the mineral soils. PMID- 19290194 TI - Ultrastructure of the Feeding Apparatus of Rhabdodemania minima Chitwood, 1936 (Enoplida: Rhabdodemaniidae). AB - The feeding apparatus of the marine nematode Rhabdodemania minima Chitwood, 1936 has been examined with light and transmission electron microscopy. The buccal capsule consists of a posterior region with smooth walls bearing three sets of three minute denticles at its posterior end and three large onchia in its mid region; a middle region with grooved walls; and an anterior region with costae and six odontia. The anterior and middle portions of the buccal capsule are enveloped by the cephalic cuticle, whereas the posterior region, which is set off from the middle region by a buccal seam, is partially enveloped by the anterior end of the esophagus. Two subventral esophageal glands open into the lumen of the esophagus. Secretions of each of three paraesophageal glands are conveyed through a duct in each of the three corresponding corners of the buccal wall to an opening between labia. A pair of wing-like thickenings, termed pterons, embraces the duct of each paraesophageal gland in the posterior and middle regions of the buccal capsule. A model of how the buccal capsule operates is proposed and tested. Morphological and functional aspects of the buccal apparatus and cephalic cuticle are compared with those of other taxa of the Enoplida, and their phylogenetic implications are discussed. PMID- 19290195 TI - Ultrastructural Cytochemistry of Secretory Granules of Esophageal Glands of Meloidogyne incognita. AB - Ultrastructural cytochemical tests for several enzymes, proteins, carbohydrates, and nucleic acids were conducted on secretory granules o pound dorsal and subventral esophageal glands of preparasitic second-stage juveniles and the dorsal gland of adult females of Meloidogyne incognita. Secretory granules in the subventral glands of juveniles stained positive for acid phosphatase. Peroxidase, DNase, RNase, cellulase, and nucleic acids were not detected in these granules. Secretory granules in the dorsal gland of adult females stained positive for peroxidase (pH 7.6) in < 50% of the tests, Acid phosphatase, beta-glucuronidase, DNase, RNase, polyphenoloxidase, cellulase, and carbohydrates were not detected in dorsal gland granules in adult females. Positive staining with cobalt thiocyanate, a stain for amino groups of basic proteins, occurred in secretory granules in the dorsal gland, ribosomes, and chromatin in adult females. Ribosomes, nuclei, and secretory granules of the dorsal gland of adult females intensely stained when incubated in three reagents specific for nucleic acid. PMID- 19290196 TI - Morphological Differences Between Radopholus citrophilus and R. similis. AB - SEM observations of the external morphology of populations of Radopholus citrophilus and R. similis revealed several diagnostic differences. The cloaco spicular orifice on males of R. citrophilus had three to seven genital papillae (anterior hypoptygmata), whereas males of R. similis were either smooth or had one or two shorter genital papillae (anterior hypoptygmata). Females of R. citrophilus had four annules in the region of the vulval opening, but R. similis had five annules in the same region. The labial disc and lateral lips appeared to be of diagnostic significance, but these areas were more susceptible to artifacts due to fixation. An unknown population of Radopholus from Puerto Rico with a chromosome number of n = 4 was morphologically similar to R. similis. These morphological differences provide additional support that R. citrophilus and R. similis are distinct species. PMID- 19290197 TI - Scanning Electron Microscopy of Vulval Cones of Heterodera glycines and Three Related Cyst Nematode Species. AB - Vulval cones of four closely related Heterodera species - H. glycines (races 1 5), H. lespedezae, H. schachtii, and H. trifolii - were examined using scanning electron microscopy. Numbers of dorsal and ventral radial ridges, total radial ridges, perineal ridges, and preanal ridges were useful in differentiating the five races of H. glycines and the other three species. Most of the populations differed significantly (P < 0.01) using the Waller-Duncan k-ratio t-test for mean separation of the five characters. H. glycines races 2, 4, and 5 were most similar. H. schachtii and H. trifolii were most dissimilar to each other and to H. lespedezae and H. glycines. Two additional qualitative characters were also useful in differentiating the populations. The shallower, shorter radial ridges of H. glycines provided a basis for separation from the other three species. Width and smoothness of the perineal ridges were useful in differentiating both races and species. PMID- 19290198 TI - First Molt in Anguina tritici. PMID- 19290199 TI - Symposium on tropical and pacific nematology-introduction. PMID- 19290200 TI - Plant-parasitic Nematode Problems in the Pacific Islands. AB - The Pacific islands have a diverse range of food and cash crops with indigenous and introduced nematode problems. The staple food crops have serious nematode pests, such as Meloidogyne spp. on sweet potato, Hirschmanniella miticausa causing corm rot of taro, and Pratylenchus coffeae and Radopholus sp. producing tuber dry rot of yams. Bananas are infested with P. coffeae or R. similis, citrus with Tylenchulus semipenetrans, rice with Aphelenchoides besseyi, and ginger with Meloidogyne spp. and R. similis. Rotylenchulus reniformis, P. zeae, P. brachyurus, and Helicotylenchus spp. are important on all of these and other crops, such as sugarcane, passion fruit, pawpaw, and cassava. Meloidogyne spp. cause serious damage to local and introduced leaf and fruit vegetables and other crops, such as tobacco, sugarcane, pawpaw, black pepper, and pyrethrum. Many other plant-parasitic genera and species, some undescribed, occur in the Pacific, and there are many islands still to be investigated. PMID- 19290201 TI - Current research on the major nematode problems in Japan. AB - AMONG IMPORTANT NEMATODE SPECIES OCCURRING IN JAPAN, CURRENT RESEARCH ACHIEVEMENTS WITH THE FOLLOWING FOUR NEMATODES ARE REVIEWED: 1) Soybean cyst nematode (SCN), Heterodera glycines - breeding for resistance, race determination, association with Cephalosporium gregatum in azuki bean disease, and isolation of hatching stimulant. 2) Potato-cyst nematode (PCN), Globodera rostochiensis - pathotype determination (Ro 1), breeding for resistance, and control recommendations. 3) Pinewood nematode (PWN), Bursaphelenchus xylophilus - primary pathogen in pine wilt disease, life cycle exhibiting a typical symbiosis with Japanese pine sawyer, Monochamus alternatus, and project for control. 4) Rice root nematodes (RRN), Hirschmanniella imamuri and H. oryzae - distribution of species, population levels in roots, and role of these nematodes in rice culture. PMID- 19290202 TI - Potential for nematode control by mycofloras endemic in the tropics. AB - Results of mycological surveys of root-knot and cyst nematodes from tropical regions indicate that most fungal species associated with females or cysts of species of Globodera, Heterodera, and Meloidogyne are those found with nematodes from temperate areas. Some fungal species, however, were found in higher frequency in tropical regions than in temperate countries; e.g., Cylindrocarpon destructans and Ulocladium atrum were the most common species associated with G. pallida and G. rostochiensis cysts in Peru. These fungi are not so frequent in nematodes from temperate areas. Fungi associated with diseased nematodes in the tropics vary greatly in nutritional requirements and include thermophilic species as well as cold-tolerant fungi. Multi-cropping systems possible in most tropical regions may be designed to increase the frequency of occurrence of microbial species antagonistic to phytonematodes. PMID- 19290203 TI - Theory and practice of the cropping systems approach to reducing nematode problems in the tropics. AB - Plant-parasitic nematodes are major constraints to the productivity of tropical farming operations. Intensive land use and climatic conditions favorable to nematode development contribute to increased crop losses due to these pests. Many farmers in developing tropical countries have limited resources and management options. Cropping systems research is a relatively low-cost, low-input method of optimizing existing agricultural practices with respect to limiting losses due to plant-parasitic nematodes. Specific tropical farming practices are discussed along with problems they pose for research in quantitative hematology. Comprehensive, systematic research methods for delineating and using nematode host relationships are described, and new ways of dealing with complex multicropping systems are suggested. PMID- 19290204 TI - Nematode problems affecting agriculture in the Philippines. AB - Nematodes are considered major pests on most economic crops in the Philippines, particularly on banana, pineapple, citrus, tomato, ramie, and sugarcane. Radopholus similis is the most destructive nematode on banana, while Meloidogyne spp. are more serious on various vegetable crops such as tomato, okra, and celery and on fiber crops such as ramie. Tylenchulus semipenetrans is a problem on citrus and Rotylenchulus reniformis on pineapple and some legume crops. Hirschmanniella oryzae and Aphelenchoides besseyi are becoming serious on rice, and Pratylenchus zeae is affecting corn in some areas. Lately, Globodera rostochiensis has been causing serious damage on potato in the highlands. Control measures such as crop rotation, planting resistant varieties, chemical nematicide application, and biological control have been recommended to control these nematodes. PMID- 19290205 TI - History of pine wilt disease in Japan. AB - Pine wilt disease induced by the pinewood nematode, Bursaphelenchus xylophilus, is a great threat to pine forests in Japan. The first occurrence of the disease was reported in Nagasaki, Kyushu. During the 1930s the disease occurrence was extended in 12 prefectures, and in the 1940s the disease was found in 34 prefectures. The annual loss of pine trees increased from 30,000 m(3) to 1.2 million m(3) during these two decades. The enormous increase in timber loss in the 1970s resulted in 2.4 million m(3) of annual loss in 1979. The affected area expanded into 45 prefectures of 47 prefectures in Japan. In cool areas the disease differs in epidemiology from that in heavily infested areas in the warm regions. A national project for controlling pine wilt disease lays special emphasis on the healthy pine forests predominating throughout cool areas in northern Japan. PMID- 19290206 TI - Nemtaode-vector relationships in the pine wilt disease system. AB - Pinewood nematode, Bursaphelenchus xylophilus, is the causal agent of pine wilt disease in North America and Japan. Dispersal stage dauer larvae are transported to new host trees on the body surface and within the tracheal system of several beetle species. Worldwide, 21 species of Cerambycidae, 1 genus of Buprestidae, and 2 species of Curculionidae are known to carry pinewood nematode dauer larvae upon emerging from nematode-infested trees. Five species of cerambycids in the genus Monochamus are known to transmit dauer larvae to new host trees, four North American species and one Japanese species. Primary transmission to healthy trees occurs through beetle feeding wounds on young branches. Secondary transmission to stressed trees or recently cut logs occurs through Monochamus oviposition sites. PMID- 19290207 TI - Pathogenesis in Pine Wilt Caused by Pinewood Nematode, Bursaphelenchus xylophilus. AB - The progression of events in the development of pine wilt disease following the invasion by Bursaphelenchus xylophilus is reviewed from early migration through pine tissues until symptom development on foliage. Disease resistance in pines, especially the hypersensitive reaction that is successful in controlling many potential pests and pathogens, is explored. Pathologies resulting from the activities of pinewood nematode include cortical trails and cavities; formation of cambial gaps and traumatic resin cysts; browning and death of cortex, phloem, cambium, and ray tissues; granulation and shrinkage of cell cytoplasm in rays; and destruction of resin canal epithelial and ray parenchyma cells. Death of parenchyma, production of toxins, and leakage of oleoresins and other material into tracheids are typical of the hypersensitive reaction occurring in pines following migration of small numbers of pinewood nematodes. The hypothesis presented is that a spreading hypersensitive reaction results in some of the observed pathologies and symptoms and eventually causes pine death. The growth differentiation balance hypothesis is used to help explain predisposition, oleoresin production and toxicity, susceptibility and resistance, and the effects of variation in climate on host pines as related to pinewilt disease. PMID- 19290208 TI - Role of phytotoxins in pine wilt diseases. AB - Characteristic rapid death of pines after infection by Bursaphelenchus xylophilus suggests the involvement of phytotoxins in the pine wilt disease syndrome. Crude extract from diseased pine is toxic to pine seedlings, whereas an extract from healthy pine is not. The response of seedlings to the crude toxin is more prominent in susceptible pine species than in resistant ones. Benzoic acid, catechol, dihydroconiferyl alcohol, 8-hydroxycarvotanacetone (carvone hydrate), and 10-hydroxyverbenone, which are toxic, low molecular weight metabolites, can be isolated from diseased pines. Other unidentified toxins are also found. The toxicity of some of these metabolites correlates positively to the susceptibility of pines to B. xylophilus. Some of these abnormal metabolites show synergistic toxicity when in combination. The D-isomer of 8-hydroxycarvotanacetone, dihydroconiferylalcohol, and 10-hydroxyverbenone inhibited the reproduction of B. xylophilus. Cellulase excreted by pinewood nematode also may be involved in rapid wilting. PMID- 19290209 TI - Carbohydrate Catabolism in Populations of Bursaphelenchus xylophilus and in B. mucronatus. AB - Genotypically different host specific pathotypes of Bursaphelenchus xylophilus have been identified. These pathotypes elicit different responses in pines depending on susceptibility, tolerance, or resistance. Continued passage of some of these pathotypes on fungal cultures leads to conversion to nonparasitic populations. These populations metabolize carbon substrates to ethanol by an anaerobic pathway, while operating some level of a phosphoenolpyruvate (PEP) succinate pathway to excrete succinate-lactate and malate. On the other hand, parasitic populations metabolize glucose to lactate-succinate, mainly by a PEP succinate pathway, and maintain redox balance through glycerol production. Ethanol and malate are not excreted by parasitic populations. PMID- 19290210 TI - Impact of pinewood nematode in north america: present and future. AB - Bursaphelenchus xylophilus, pinewood nematode (PWN), is the most serious pest of pine forests in Japan, but in North America its role in pine wilt disease is still being studied. The PWN is known to infest many species of Pinus, with P. nigra, P. sylvestris, and P. thunbergii the most susceptible in the eastern United States. Because of its potential, several European countries (Finland, Norway, and Sweden) and Korea have established embargoes against the importation of coniferous wood from regions of the world known to be infested with the PWN. Although the PWN is not considered an economic pest in North American forests, the recent embargoes have established an impact on current forest management practices and an economic impact on North American export trade. PMID- 19290211 TI - Tylenchulus graminis n. sp. and T. palustris n. sp. (Tylenchulidae), from Native Flora of Florida, with Notes on T. semipenetrans and T. furcus. AB - Tylenchulus graminis n. sp. and T. palustris n. sp. are described and illustrated from broomsedge (Andropogon virginicus L.) and pop ash (Fraxinus caroliniana Mill.), respectively. T. graminis resembles T. furcus in having a distinct anus, but T. graminis second-stage juveniles (J2) do not have a bifid tail. T. semipenetrans does not have a perceptible anus. The mature female of T. graminis has a mucronate pointed terminus while T. semipenetrans has a smooth and round terminus. T. graminis males have wider stylet knobs and basal bulb and a longer tail than T. semipenetrans males. T. graminis J2 have a longer posterior body portion (without large fat globules) than T. semipenetrans J2. T. palustris resembles T. semipenetrans in having an undetectable anus but differs by the short and conoid mature female postvulval section. The male of T. palustris has larger stylet knobs and basal bulb than those of T. semipenetrans and a bluntly rounded tail terminus, which is tapered in T. semipenetrans. T. palustris differs from T. furcus and T. graminis in having an undetectable anus, by the conoid postvulval section of mature females, by the shorter and rounded tail of males, and the shorter J2 posterior body section without large fat globules. T. graminis and T. palustris are parasites of indigenous flora of Florida. PMID- 19290212 TI - Two new species of prismatolaimus de man, 1880 (nemata: prismatolaimidae) in southern chile. AB - Two new species of Prismatolaimus are described from Orange Bay, Hoste Island, Chile. Prismatolaimus novoporus sp. n. is distinguished by its females being 1.08 1.28 mm long and monovarial with a long postvulvar sac, and having a long tail (c' = 21.5-32.2); males have 11-20 supplements confined to the posterior part of the body. Prismatolaimus chilensis sp. n. is distinguished by its amphidelphic reproductive system, short tail (c' = 9.5), V = 63.4; males have 23 supplements reaching up to the neck region. A well-developed dorsal body pore behind the cardia connected with an apparently glandular organ is reported in P. novoporus sp. n. Function of the organ is unknown, and it is not a general feature in the genus. The generic diagnosis of Prismatolaimus is emended and keys to species, both females and males, are presented. The systematic position of Prismatolaimus is discussed, and it is judged to be the only genus of Prismatolaimidae. Also, it is concluded that Prismatolaimidae and Bastianidae represent two sister taxa in Araeolaimida or Leptolaimina. PMID- 19290213 TI - Transmission of Tomato Ringspot Virus by Xiphinema americanum and X. rivesi from New York Apple Orchards. AB - Populations of Xiphinema americanum and X. rivesi were collected from apple orchards in eastern and western New York and tested in the laboratory for ability to transmit tomato ringspot virus (TmRSV) to cucumber and dandelion. Populations varied in the frequency with which they transmitted TmRSV, but this variation did not correspond to variation in disease prevalence in the orchard. The lower prevalence of TmRSV-incited disease in apple trees in western New York cannot be attributed to inability of the local Xiphinema spp. to transmit TmRSV. PMID- 19290214 TI - Genomic Differences among Pathotypes of Bursaphelenchus xylophilus. AB - Total genomic DNA from Bursaphelenchus xylophilus pathotypes MPSy-1 and VPSt-1 and from B. mucronatus was digested with restriction endonucleases. DNA fragments were electrophoretically separated, Southern blotted to nitrocellulose, and hybridized to genomic DNA from one of the isolates. The resulting hybridization patterns indicate genomic differences in repetitive DNA sequences among these populations. Greatest differences were seen between B. xylophilus and B. mucronatus, but genomic differences were also apparent between B. xylophilus pathotypes MPSy-1 and VPSt-1 and between a population from P. nigra in New Jersey and a population of a mucronate form from Abies balsamea in Quebec, Canada. PMID- 19290215 TI - Field Evaluation of Steinernema feltiae Against the Web-spinning Larch Sawfly Cephalcia lariciphila. AB - Field trials were conducted in Rheola Forest, Wales, Great Britain, to determine the effectiveness of Steinernema feltiae UK strain in controlling the web spinning larch sawfly Cephalcia lariciphila. Foliar sprays at the rate of 5,000 20,000 nematodes/100 cm branch resulted in 3.4-29.4% infection of sawfly larvae. Soil application of 200 nematodes/cm(2) resulted in 61% infection of sawfly prepupae and 17.3% of pupae. Prepupal infection ranged from 4.8 to 14.7% 1 year after nematode application. Soil applications of this nematode show that it has potential for biological control of sawfly prepupae. PMID- 19290216 TI - Lectin binding sites on the amphidial exudates of meloidogyne. AB - Lectin binding sites on the surface of Meloidogyne incognita Races 1, 2, 3, and 4; M. javanica; M. arenaria Races 1 and 2; and M. hapla Races A and B were determined with lectins conjugated to fluorescein isothiocyanate or colloidal gold. The amphidial exudate, which was demonstrated histochemically to contain carbohydrate, was the principal binding site. Some lectins also bound to the external cuticular surface. Species and race specific binding patterns were observed for both amphidial and cuticular binding sites. PMID- 19290217 TI - Cephalobus litoralis: Biology and Tolerance to Desiccation. AB - Cephalobus litoralis (Akhtar, 1962) Andrassy, 1984 reproduced parthenogenetically and completed its life cycle in 72-90 hours. Each female deposited 200-300 eggs. The nematodes showed synchronized movements in the rhythms of the anterior parts of the body. The nematodes were coiled when dried in culture medium or in slowly evaporating water droplets on the tops of culture plates, but in pellets they assumed irregular postures. Nematodes in pellets stored at high humidity could be reactivated after storage for 28 days. PMID- 19290218 TI - Soybean Response to Two Isolates of Meloidogyne arenaria. PMID- 19290219 TI - Multiple focus and exposure photomicroscopy of nematodes for increased depth of field. PMID- 19290220 TI - Effects of Host Resistance on Second-stage Juveniles and Adult Males of Globodera rostochiensis. AB - Potato cultivars Katahdin (susceptible) and Rosa (resistant) were exposed to infective second-stage juveniles (J2) of Globodera rostochiensis for varying periods of time, after which root systems were washed and plants were placed in Hoagland's solution to assess J2 egression and male emergence. After transfer to liquid culture, many J2 egressed from both cultivars, but significantly more egressed from the resistant Rosa than from Katahdin. Juveniles that egressed from Rosa invaded a second host, resistant or susceptible, in significantly fewer numbers than did juveniles that egressed from Katahdin. Also, significantly fewer males developed in and emerged from resistant host roots, relative to susceptible ones. These effects of resistance may be an important component of the tolerance to invasion by G. rostochiensis exhibited by Rosa. PMID- 19290221 TI - Age structure and community diversity of nematodes associated with maize in iowa sandy soils. AB - Age structure of nematode populations around maize growing in sandy soils in Iowa was studied at soil depths of 0-15and 15-30 cm for 2 years. Numbers of Longidorus breviannulatus were generally greater at 0-15 cm than at 15-30 cm deep until mid to late season. The decline in numbers of females as the season progressed indicates that fecundity slowed and is evidence of only one generation per year. Peak populations of Pratylenchus scribneri and Xiphinema americanum occurred in late August or early September. Adults of Hoplolaimus galeatus were few in the roots but common in the soil, indicating that fertilization occurred mostly in the soil. Numbers of P. scribneri were generally greater at the lower depth, especially late in the season. Community diversity (H') was less when nematode biomass was used instead of numbers. Numbers of H. galeatus did not decline over the winter. Numbers of L. breviannulatus, P. scribneri, and X. americanum declined significantly over the winter, but not between spring cultivation and planting. PMID- 19290222 TI - Behavioral Responses of Meloidogyne incognita to Small Temperature Changes. AB - Small, rapid temperature changes were generated by incandescent radiation, and behavioral responses of Meloidogyne incognita juveniles were recorded with high time resolution by computer tracking. Temperature changes away from the preferred temperature resulted in decreases in the rate of movement and increases in the rate of change of direction, whether the changes were toward warmer or cooler temperatures. These behavioral changes lasted about 30 seconds. Temperature changes toward the preferred temperature caused the response rates to change in the opposite directions, and the behavioral changes persisted for several minutes. These results demonstrate that nematodes can respond to a purely temporal thermal stimulus in a manner consistent with efficient indirect orientation or klinokinesis. The rate of temperature change was estimated to be of the order of 10 C/second, suggesting that the nematodes detected a change of about 0.001 C. PMID- 19290223 TI - Effect of Temperature on Infection and Survival of Rotylenchulus reniformis. AB - From infestation of lettuce with preinfective females to egg deposition, populations of Rotylenchulus reniformis from Baton Rouge, Louisiana; Lubbock and Weslaco, Texas; and Mayaguez, Puerto Rico, required 41, 13, 7, and 7 days at 15, 20, 25, and 34 C, respectively. No nematode infection occurred at 10 C with any R. reniformis population, and the population from Puerto Rico did not reproduce at 15 C. Nematode survival was not influenced by temperature, since populations from Texas and Louisiana survived for 6 months without a host at - 5 , - 1 , 4, and 25 C. Survival of R. reniformis was substantially influenced by soil moisture. Soil moistures greater than 7% (< 1 bar) aided nematode survival at storage temperature of 25 C, whereas moisture adversely affected nematode survival below freezing. Soil moisture below 4% (> 15 bars) favored nematode survival below freezing but adversely affected nematodes in soils stored at 25 C. Soil moisture effects on nematode survival were less accentuated at 4 and 0 C. PMID- 19290224 TI - Histology of the Interactions of Paecilomyces lilacinus with Meloidogyne incognita on Tomato. AB - Excised tomato roots were examined histologically for interactions of the fungus Paecilomyces lilacinus and Meloidogyne incognita race 1. Root galling and giant cell formation were absent in tomato roots inoculated with nematode eggs infected with P. lilacinus. Few to no galls and no giant-cell formation were found in roots dipped in a spore suspension of P. lilacinus and inoculated with M. incognita. Numerous large galls and giant cells were present in roots inoculated only with M. incognita. P. lilacinus colonized the surface of epidermal cells as well as the internal cells of epidermis and cortex. The possibility of biological protection of plant surfaces with P. lilacinus against root-knot nematodes is discussed. PMID- 19290225 TI - Development of Heterodera glycines Life Stages as Influenced by Temperature. AB - The effects of temperature on rates of development of Heterodera glycines egg and juvenile stages were examined as a basis for predicting generation times of the nematode on soybean. The relationship of temperature to H. glycines embryonic development between 15 and 30 C was described by a linear model, The calculated basal temperature threshold was 5 C. Thermal optimum for embryogenesis and hatch with low mortality was 24 C. Development proceeded to first-stage juvenile at 10 C and to second-stage juvenile at 15-30 C. Hatch occurred at 20-30 C. At 36 C, development proceeded to the four-cell stage, then the eggs died. The range of diurnal soil temperature fluctuation and accumulated degree-days between 5 and 30 C (DD5/30) had an impact on rate of development of juveniles in soybean roots. From early June to early July, H. glycines required 534 + 24 DD5/30 (4 weeks) to complete a life cycle in the field. During the midseason (July and August), life cycles were completed in 3 weeks and 429 +/- 24 DD5/30 were accumulated. Late in the season (September to November), declining soil temperatures were associated with generation times of 4 weeks and slower rates of development. PMID- 19290226 TI - Effect of Within-field Variation in Soil Texture on Heterodera glycines and Soybean Yield. AB - The influence of soil texture on Soybean yield in the presence of Heterodera glycines was investigated by comparing yields of susceptible cultivars with a resistant cultivar for 2 years. Soybean yield was negatively correlated with increasing sand content (P = 0.05). Yields of susceptible cultivars were suppressed with increasing sand content. Final nematode population densities were lowest in plots with greatest sand content. Soybean infection by SCN, as determined by the number of cysts 30 days after planting, was not consistently related to soil texture over 2 years. Initial nematode population density was positively related to soybean yield the first year and negatively related to soybean yield the second, probably a result of greater yield suppression by H. glycines in plots with greater sand content. PMID- 19290227 TI - Redescription of Heterodera fici (Nematoda: Heteroderidae) with SEM Observations. AB - Heterodera fici is redescribed and illustrated with comparative details and revised measurements and diagnostic characters of the females, males, cysts, and juveniles from Maryland and Pakistan. This species is in the "schachtii group" (cysts lemon shaped, with bullae, and ambifenestrate) but the fenestrae in some cysts, presumab!y young ones, are small and widely spaced, appearing bifenestrate. It is most closely related to H. schachtii, H. glycines, and H. cajani but differs from these species especially in having cysts with small, scattered bullae and weakly developed underbridge; and males with four small nipples on tail tip. Scanning electron microscopy (SEM) observations of the specimens are also presented. The relationship of this species to closely related forms is discussed. PMID- 19290228 TI - Complete Characterization of the Race Scheme for Heterodera glycines. AB - One hundred thirty-eight isolates of Heterodera glycines from nine states in the United States, People's Republic of China, and Indonesia were tested on the four standard soybean race differentials. A total of 12 variants were found, including the five races described previously. The seven variants that did not correspond to one of the described races and reports from other areas of populations that could not be classified are evidence that the present race classification system needs to be fully characterized. Eleven additional races are described; this expands the total to 16 races, the maximum possible using the four prescribed differentials and a + or - rating for each. The seven new races are designated as 6, 9, 10, 13, 14, 15, and 16. This complete characterization of the race scheme will allow for immediate communication of the discovery of the remaining four races plus the identification of previous undescribed races. PMID- 19290229 TI - Factors Affecting the Biology and Pathogenicity of Heterodera schachtii on Sugarbeet. AB - A direct relationship exists between soil temperature and Heterodera schachtii development. The average developmental period of two nematode populations from Lewiston, Utah, and Rupert, Idaho, from J2 to J3, J4, adult, and the next generation J2 at soil temperatures of 18-28 C were 100, 140,225, and 399 degree days (base 8 C), respectively. There was a positive relationship (P < 0.05) between nematode Pi, nematode generations, and sugarbeet yields. The greatest sugarbeet growth inhibition (87%) occurred when sugarbeets were exposed to a Pi of 12 eggs/cm(3) soil for five generations (1,995 degree-days), compared with a 47% inhibition when plants were exposed to the same Pi for two generations. There was a negative correlation (P < 0.05) between the Pi, Pf, and sugarbeet yield for each population threshold. The smaller the Pi, the greater the sugarbeet yields and the greater the Pf. Root yields were 80 and 29 t /ha and Pf were 8.4 and 3.6 eggs/cm(3) soil when sugarbeet seeds were planted at Pi of 0.4 and 7.9 eggs/cm(3). respectively, at a soil temperature of 8 C. The number of years rotation with a nonhost crop required to reduce the nematode population density below a damage threshold level of 2 eggs/cm(3) depends on the Pi. A Pi of 33.8 eggs/cm(3) soil required a 5-year crop rotation, whereas a Pi of 8.4 eggs/cm(3) soil required a 2-year crop rotation. PMID- 19290230 TI - Growth of Potato and Control of Pratylenchus penetrans with Oxamyl-treated Seed Pieces in Greenhouse Studies. AB - Oxamyl was applied to both uncut and cut potato tubers in aqueous solutions of 1,000 to 32,000 mug/ml. Emergence in greenhouse pots was delayed for a day or more after soaking cut tuber pieces in 32,000 mug/ml. After 10 weeks plant growth was greater, relative to the control, when Pratylenchus penetrans-infested soil was planted with cut tubers soaked for 20 minutes in 32,000 mug/ml. Soaking for 40 minutes did not increase nematode control nor affect plant growth. Oxamyl applied to tubers at 1,000 mug/ml reduced the numbers of P. penetrans in the soil by 20% and in the roots by 35%; at 32,000 mug/ml, the numbers of P. penetrans in the soil were reduced by 73-86% and in the roots by 86-97%. The numbers of P. penetrans did not increase in the roots of plants developed from cut tubers soaked in 32,000 mug/ml over a period of 10 weeks, but numbers of lesion nematodes had begun to increase in the soil. PMID- 19290231 TI - Differential Response of Thor Alfalfa to Meloidogyne chitwoodi Races and M. hapla. AB - Second-stage juveniles (J2) of races 1 and 2 of Meloidogyne chiiwoodi and M. hapla readily penetrated roots of Thor alfalfa and Columbian tomato seedlings; however, few individuals of M. chitwoodi race 1 were able to establish feeding sites and mature on alfalfa. Histopathological studies indicate that J2 of race 1 either failed to initiate feeding sites or they caused cell enlargement without typical cell wall thickening. The protoplasm of these cells coagulated, and juveniles of race 1 did not develop beyond the swollen J2 stage. A few females of race 1 fed on small giant cells and deposited a few eggs at least 20 and 30 days later than M. chitwoodi race 2 and M. hapla, respectively. Failure of race 1 to establish feeding sites was related to egression of J2 from the roots. The M. chitwoodi race 1 J2 egression from alfalfa roots was higher than egression of race 2 and M. hapla. Egression of J2 of M. chitwoodi races 1 and 2 from tomato roots was similar and higher than that of M. hapla. Thus egression plays an important role in the host-parasite relationship of M. chitwoodi and alfalfa. PMID- 19290232 TI - Variability in Host Preference Among Field Populations of Heterodera glycines. AB - Eighteen soybean fields, six each with race 3, race 4, or Bedford population of Heterodera glycines, were selected for testing of host variability. Each field was divided into three sections, and a bulk soil sample was taken from each section. The 54 bulk soil populations (BSP) and 270 single cyst populations (SCP) were subjected to race determination tests. Tests of the 18 BSP and 90 SCP from the race 3 fields revealed that race 3 was the predominant race; however, 68 of the populations tested were other races. Tests of the 18-BSP and 90 SCP from race 4 fields demonstrated that races 2 and 4 were predominant, with 38 and 39 populations; respectively. Tests of the 18 BSP and 90 SCP from the Bedford population fields revealed tremendous variability. Races 2, 4, and 6 were the predominant races, with 32, 31, and 28 populations, respectively. These results indicate that of the three races studied, the Bedford population is the most variable, race 3 shows considerable variability, and race 4 shows very little. PMID- 19290233 TI - Growth and yield responses of soybean to aldicarb. AB - A series of greenhouse, phytotron, field, and microplot experiments evaluated factors that influenced plant-growth.stimulation associated with the use of the pesticide aldicarb. A phytotron experiment showed.that aldicarb increased growth, of Ransom soybean at all temperatures but was somewhat phytotoxic to Coker 156 soybean at 30 C. Soybean gave the greatest response to this nematicide at 22 C in a commercially available medium, Metromix 220. Soybean cultivars Ransom and Coker 156. exhibited increased growth in response to aldicarb or, to a lesser extent aldicarb sulfone treatments under greenhouse and microplot conditions. Enhanced soybean growth, however, did not always result in significantly greater soybean seed yield. Soil type affected soybean sensitivity to aldicarb, with. the greatest growth and yield increases generally occurring in fine-textured soils or those with high.organic matter. Plant-growth stimulation by aldicarb occurs in the absence of pests but is dependent upon concentration and edaphic and other environmental factors. PMID- 19290234 TI - Influence of aldicarb on the growth and yield of tobacco. AB - Microplot and field experiments were used to examine the plant-growth stimulation frequently associated with the use of aldicarb on tobacco in the absence of major pests. Aldicarb rates of 1.5-4.5 kg a.i./ha enhanced tobacco growth and yield in most experiments, but higher rates (>/= 4.5 kg) usually resulted in a neutral to negative effect. Tobacco cultivars NC 82 and Speight G-28 were more responsive than McNair 944 to the pesticide in microplots. Supplemental irrigation enhanced the responsiveness of Speight G-70 tobacco and McNair 944 to aldicarb, but excessive moisture (ca. 7-8 cm/week) limited cured-leaf yields. Aldicarb also resulted in the greatest mean tobacco yields in 35 field experiments involving Meloidogyne spp. over 3 years, relative to ethoprop, 1,3-dichloropropene (1,3-D), 1,3-D + chloropicrin, and nontreated controls. Thus, aldicarb generally enhanced tobacco growth and yield in the presence or absence of nematodes, but its impact is dependent on other variables, including cultivar, soil moisture, and soil type. PMID- 19290235 TI - Relationship Between Time of Infection with Heterodera glycines and Soybean Yield. AB - Experiments were conducted to determine the relationship between time of infection by Heterodera glycines and soybean growth in the greenhouse and yield of plants grown in the field. Soybean cultivar Essex seedlings growing in the greenhouse were inoculated with H. glycines at 2, 4, or 6 weeks after planting. Seedling growth was inhibited by H. glycines infection at 2 or 4 weeks after planting but not at 6 weeks. Infection of Essex by H. glycines in the field was delayed 2-6 weeks by nematicides. Yields were significantly increased when H. glycines infection was delayed 2 weeks by nematicide treatment. Essex yields were highest when infection was delayed 6 weeks, equalling the yield of the H. glycines-resistant cultivar Asgrow 5474. The effect of H. glycines on soybean growth in the greenhouse and yields in the field declined when infection was delayed 6 weeks. Thus, soybean sensitivity to H. glycines seemed to diminish with age of the soybean plants. PMID- 19290236 TI - Comparisons of Mitochondrial DNA from the Sibling Species Heterodera glycines and H. schachtii. AB - Restriction fragment patterns of mitochondrial DNA from sibling species of cyst nematodes Heterodera glycines and H. schachtii were examined. Fourteen restriction endonucleases recognizing four, five, and six base-pair sequences yielded a total of 90 scorable fragments of which 10% were shared by both species. Mitochondrial genome sizes for H. glycines and H. schachtii were estimated to be 22.5-23.5 kb and 23.0 kb, respectively. A single wild type mitochondrial genome was identified in all populations of H. glycines examined, although other mitochondrial genomes were present in some populations. The H. schachtii genome exhibited 57 scorable fragments, compared with 33 identified in the H. glycines wild type genome. The estimated nucleotide sequence divergence between the two species was p = 0.145. This estimate suggests these species diverged from a common ancestor 7.3-14.8 million years ago. PMID- 19290237 TI - Meloidogyne incognita and Tomato Response to Thiamine, Ascorbic Acid, L-arginine, and L-glutamic Acid. AB - The influence of solutions of ascorbic acid, thiamine, L-arginine, and L-gtutamic acid on egg hatch, juvenile survival, and development and reproduction of Meloidogyne incognita in susceptible and resistant tomatoes was studied. Maximum inhibition of egg hatch occurred at 2,000, 4,000, and 2,000 ppm for ascorbic acid, L-arginine, and L-glutamic acid, respectively. Larval survival was significantly reduced by concentrations of 2,000 ppm ascorbic acid and 1,000 ppm of L-arginine. Maximum inhibition of egg hatch and mortality of juveniles was achieved at a concentration of 4,000 ppm of ascorbic acid and L-arginine. L glutamic acid and thiamine had respective moderate and minimal toxic effects. Foliar sprays of ascorbic acid, L-arginine, or L-glutamic acid suppressed the numbers of root galls, females, and egg masses on the susceptible tomato cultivar Tropic. Ascorbic acid and L-arginine had highly significant effects when applied to foliage before inoculation with nematodes. Thiamine had little effect. All sprays suppressed the numbers of root galls and females in roots of the resistant cultivar VFN8 when treatments were applied before inoculation. They were not, however, effective as post-inoculation treatments. Growth of a susceptible cultivar was improved by post-inoculation and pre-inoculation treatments when compared with the control plants which had neither nematode infection nor chemical treatment. No positive growth response to chemical treatment was seen in resistant control plants. PMID- 19290238 TI - Influence of Calonectria crotalariae on Reproduction of Heterodera glycines on Soybean. AB - Calonectria crotalariae enhanced root penetration of Lee 74 (susceptible) and Centennial (resistant) soybeans by juveniles of race 3 of Heterodera glycines. Numbers of cysts in and on the roots of Lee 74 increased during the first 30 days in the presence of the fungus. Percentage of root infection by the fungus increased at 40 days in Lee 74 in the presence of the nematode. Numbers of cysts in soil at 80 and 120 days after inoculation with both organisms accounted for the significantly increased nematode population levels on Lee 74. In the presence of the fungus on the resistant cultivar, significantly increased levels of cysts were recovered from soil at 120 days. Fungus infection of Centennial roots also infected with the nematode increased from 58 to 86% at 120 days. An inoculum timing study in which Lee 74 was infested with the nematode and fungus individually, sequentially, and in combination at days 0 and 35 indicated that enhanced nematode reproduction was related more to early plant-fungus than to early plant-fungus-nematode interaction(s). PMID- 19290239 TI - Host Tests to Differentiate Meloidogyne chitwoodi Races 1 and 2 and M. hapla. AB - The reproductive factor (R = final egg density at 55 days / 5,000, initial egg density) of Meloidogyne chitwoodi race 2 (alfalfa race) on 46 crop cultivars ranged from 0 to 130. The reproductive efficiency of M. chitwoodi race 1 (non alfalfa race) and M. chitwoodi race 2 was compared on selected crop cultivars. The basic difference between the two races lay in their differential reproduction on Thor alfalfa and Red Cored Chantenay carrot. M. chitwoodi race 2 reproduced on alfalfa but not on carrot. Conversely, alfalfa was a poor host and carrots were suitable for M. chitwoodi race 1. Based on host responses to M. chitwoodi races and M. hapla, a new differential host test was proposed to distinguish the common root-knot nematode species of the Pacific Northwest. PMID- 19290240 TI - Effect of Three Plant Species on Population Densities of Xiphinema americanum and X. rivesi. AB - A taxonomic revision of the Xiphinema americanum species complex has necessitated a reexamination of the host range of species in the complex before recommendations can be made with confidence on the likelihood that specific crops will be damaged. Toward this end, populations of X. americanum and X. rivesi collected from apple orchards in eastern and western New York state were evaluated after 3 months in pots planted with cucumber, apple, or dandelion seedlings. Eastern and western New York populations of both nematode species declined on cucumber but increased to similar final densities on apple and dandelion. PMID- 19290241 TI - An alternative gelling agent for culture and studies of nematodes, bacteria, fungi, and plant tissues. AB - Pluronic F127 polyol, a block copolymer of propylene oxide and ethylene oxide, was studied as an alternative to agar in culture media for nematodes, bacteria, fungi, actinomycetes, and plant tissues or seedlings, At a polyol concentration of 20% w/v, the culture media, semi-solid at room temperature (22 C) but liquid at lower temperatures, had minimal effects on the test organisms. Most of the fungi and bacteria grew as well in 20% polyol as in 1.5% agar media; however, various species of nematodes and plant seedlings or tissues exhibited differential sensitivities to different concentrations of the polyol. In cases where the organisms were unaffected, the polyol media had certain advantages over agar, including greater transparency and less contamination under nonaseptic conditions. Polyol media have potentially greater ease for recovery of embedded organisms or tissues inside the media by merely shifting to lower temperatures. PMID- 19290242 TI - Genetic Analysis of Esterase Polymorphism in the Soybean Cyst Nematode, Heterodera glycines. AB - The genetic basis of esterase polymorphism in Heterodera glycines was investigated through controlled matings and analysis of F and F progeny. Three nematode lines, each fixed for a different esterase phenotype, were isolated and purified through repeated directional selection and inbreeding. Each phenotype was characterized by its distinct pair of closely spaced bands of esterase activity. Single-female single-male crosses were conducted according to a modified agar-plate mating technique. F progeny were homogeneous, exhibiting both parental esterase phenotypes (codominant heterozygotes) but no hybrid bands. Approximately 1,500 F progeny segregated in a 1:2:1 ratio for expression of the esterase phenotypes of the female parental line, the heterozygote, and the male parental line. Apparently the three esterase phenotypes correspond to three codominant alleles of a single esterase locus. Reciprocal crosses gave similar results, suggesting no maternal inheritance. PMID- 19290243 TI - A role for the 'excretory' system in secernentean nematodes. PMID- 19290244 TI - A Simple Method for Examination of the Vulva Area of Mature Cysts of Heterodera spp. PMID- 19290245 TI - Wheat Germ Agglutinin Binding to the Outer Cuticle of the Plant-parasitic Nematode Anguina tritici. PMID- 19290246 TI - A Simple Technique for Mounting Whole Root-knot Nematode Females. PMID- 19290247 TI - Estimation of genetic divergence in meloidogyne mitochondrial DNA. AB - Restriction fragments from purified mitochondrial DNA can be readily detected following rapid end-labeling with [alpha-(3)(2)]nucleoside triphosphates and separation by gel electrophoresis. Mitochondrial DNA from 12 populations of Meloidogyne species was digested with 12 restriction enzymes producing more than 60 restriction fragments for each species. The mitochondrial genome of M. arenaria is the most genetically distinct of the four species compared. M. arenaria shows approximately 2.1-3.1% nucleotide sequence divergence from the mitochondrial genomes of M. javanica, M. incognita, and M. hapla. Among the latter three species, interspecific estimates of sequence divergence range from 0.7 to 2.3%. Relatively high intraspecific variation in mitochondrial restriction fragment patterns was observed in M. hapla. Intraspecific variation in M. incognita resulted in sequence divergence estimates of 0.5-1.0%. Such polymorphisms can serve as genetic markers for discerning mitochondrial DNA genotypes in nematode populations in the same way that allozymes have been used to discern nuclear DNA genotypes. PMID- 19290248 TI - Relationship Between Egg Viability and Population Densities of Meloidogyne incognita on Cotton. AB - Cotton seedlings were inoculated with a range of initial populations (Pi) of Meloidogyne incognita in greenhouse experiments to test the relationship between nematode population densities and egg viability. In two of three experiments, a significant (P < 0.05) negative linear relationship was detected between percentage of hatch of first generation eggs and log Pi. A similar relationship between hatch and root-gall index was observed. In two experiments numbers of eggs judged to be nonviable based on appearance were significantly greater (P < 0.05) in the highest Pi (60,000 eggs/seedling) treatments than in treatments with lower Pi (600-6,000 eggs/seedling). It was concluded that Pi affects egg viability measured as percentage of hatch and that this relationship may play a role in the density-dependent winter survival rates of Meloidogyne species. PMID- 19290249 TI - Effect of Meloidogyne incognita and M. javanica on Leaf Water Potential and Water Use of Tobacco. AB - Greenhouse lysimeter and field microplot tests were conducted to evaluate the effects of Meloidogyne incognita and M. javanica on plant water relations and growth performance of NC 2326 flue-cured tobacco. In the greenhouse, afternoon leaf water potential values at 8-11 weeks after transplanting were lower by as much as 0.22 MPa in plants infected with either nematode than in the control plants. From 11 to 22 weeks, leaf water potential values were similar in all treatments. Over the course of the 22-week experiment, all infected plants showed similar evapotranspiration patterns, and plants in these treatments used 87-88% of the water utilized by noninfected plants. Biomass production from nematode infected plants, however, was only about 50% of the biomass of control plants. The field microplot study showed water use patterns similar to those in the lysimeter study. PMID- 19290250 TI - Nematode Mitochondrial DNA: Anomalies and Applications. AB - The mitochondrial genome of animal cells is currently under intense investigation by molecular, evolutionary, and population biologists. This review summarizes the available information on the molecular biology of nematode mitochondrial DNA and explains how the fundamental knowledge obtained from these basic studies may be applied to nematode systematics, evolution, and diagnostics. PMID- 19290251 TI - DNA Restriction Fragment Length Polymorphism in Races of the Soybean Cyst Nematode, Heterodera glycines. AB - Restriction endonuclease digests of total DNA from races 3, 4, and 5 of the soybean cyst nematode, Heterodera glycines, have been analyzed on agarose gels. DNA fragment patterns of race 4 were completely different from those patterns obtained for races 3 and 5 by all eight restriction enzymes tested. Differences in long and short restriction DNA fragments generated by the enzyme Msp I or its isoschizomer, Hpa II, were detected between race 3 and 5 digestion profiles. Rapid DNA isolation followed by its digestion with either Msp I or Hpa II enzymes and visualization of repetitive DNA fragments in agarose gels provided a diagnostic assay for the populations of the three races examined in this study. PMID- 19290252 TI - Influence of mycorrhizal fungus, phosphorus, and burrowing nematode interactions on growth of rough lemon citrus seedlings. AB - Rough lemon seedlings were grown in mycorrhizal-infested or phosphorus-amended soil (25 and 300 mg P/kg) in greenhouse experiments. Plants Were inoculated with the citrus burrowing nematode, Radopholus citrophilus (0, 50, 100, or 200 nematodes per pot). Six months later, mycorrhizal plants and nonmycorrhizal, high P plants had larger shoot and root weights than did non-mycorrhizal, low-P plants. Burrowing nematode population densities were lower in roots of mycorrhizal or nonmycorrhizal, high-P plants than in roots of nonmycorrhizal, low P plants; however, differences in plant growth between mycorrhizal and nonmycorrhizal plants were not significant with respect to initial nematode inoculum densities. Phosphorus content in leaf tissue was significantly greater in mycorrhizal and nonmycorrhizal, high-P plants compared with nonmycorrhizal, low-P plants. Nutrient concentrations of K, Mg, and Zn were unaffected by nematode parasitism, whereas P, Ca, Fe, and Mn were less in nematode-infected plants. Enhanced growth associated with root colonization by the mycorrhizal fungus appeared to result from improved P nutrition and not antagonism between the fungus and the nematode. PMID- 19290253 TI - Growth and Energy Demand of Meloidogyne incognita on Susceptible and Resistant Vitis vinifera Cultivars. AB - Food (energy) consumption rates ofMeloidogyne incognita were calculated on Vitis vinifera cv. French Colombard (highly susceptible) and cv. Thompson Seedless (moderately resistant). One-month-old grape seedlings in styrofoam cups were inoculated with 2,000 or 8,000 M. incognita second-stage juveniles (J2) and maintained at 17.5 degree days (DD - base 10 C)/day until maximum adult female growth and (or) the end of oviposition. At 70 DD intervals, nematode fresh biomass was calculated on the basis of volumes of 15-20 nematodes per plant obtained with a digitizer and computer algorithm. Egg production was measured at 50-80 DD intervals by weighing 7-10 egg masses and counting the number of eggs. Nematode growth and food (energy) consumption rates were calculated up to 1,000 DD based on biomass increase, respiratory requirements, and an assumption of 60 % assimilation efficiency. The growth rate of a single root-knot nematode, excluding egg production, was similar in both cultivars and had a logistic form. The maximum fresh weight of a mature female nematode was ca. 29-32 mug. The total biomass increase, including egg production, also had a logistic form. Maximum biomass (mature adult female and egg mass) was 211 mug on French Colombard and 127 mug on Thompson Seedless. The calculated total cost to the host for the development of a single J2 from root penetration to the end of oviposition for body growth and total biomass was 0.535 and 0.486 calories with a total energy demand of 1.176 and 0.834 calories in French Colombard and Thompson Seedless, respectively. PMID- 19290254 TI - Host-parasite Relationship of Carrot Cultivars and Meloidogyne chitwoodi Races and M. hapla. AB - Most of the 15 carrot cultivars tested were moderate to good hosts to Meloidogyne chitwoodi race 1, whereas all except Orlando Gold were nonhosts or poor hosts for M. chitwoodi race 2. All carrot cultivars were good hosts for M. hapla. The plant weights of the carrot cultivars Red Cored Chantenay and Orlando Gold infected with either race of M. chitwoodi were significantly less than uninoculated checks in pots. Under field microplot conditions, however, detrimental effects on quality were rarely observed. M. hapla was pathogenic to both cultivars in the greenhouse and the field. The tolerance level of Orlando Gold to M. hapla was lower than Red Cored Chantenay. PMID- 19290255 TI - Variability in Race Tests with Heterodera glycines. AB - Tests of Heterodera glycines on differential host plants to determine races were run in Arkansas, Illinois, and North Carolina to check the uniformity of results of the test. Methods used at the three locations varied somewhat. Results indicate that the race test is highly variable. Isolates previously identified as race 1 were identified as race 1 or race 3; those identified as race 2 were identified in these tests as race 2, 4, 9, or 14; those previously identified as race 3 were identified as race 1 or race 3; those identified as race 4 were identified in these tests as race 4 or race 14; those previously identified as race 5 were identified as race 2; and those previously identified as race 6 were identified as race 1, 2, 4, 5, or 6. Part of the variability resulted from the use of differential host plants from different sources and part from nonstandard differential host plants. Other variations may be due to inability to obtain completely uniform inoculum or to recover all nematodes that penetrated. PMID- 19290256 TI - Description of the Male of Ogma cobbi and Juveniles of Bakernema inaequale (Nematoda: Criconematidae). AB - Juveniles ofBakernema inaequale and the male of Ogma cobbi are described and illustrated for the first time. Juveniles of Bakernema inaequale lack the cuticular spines characteristic of the adults. The annules of first-stage and fourth-stage juveniles are smooth, whereas those of the second-stage and third stage juveniles have faint, bead-like ornamentation either throughout the body or in the anterior or posterior part only. Detailed morphometric data of O. cobbi males and all juvenile stages and females of B. inaequale are given. PMID- 19290257 TI - Evaluation of Paecilomyces lilacinus as a Biocontrol Agent of Meloidogyne javanica on Tobacco. AB - The efficacy of the nematode parasite Paecilomyces lilacinus, alone and in combination with phenamiphos and ethoprop, for controlling the root-knot nematode Meloidogyne javanica on tobacco and the ability of this fungus to colonize in soil under field conditions were evaluated for 2 years in microplots. Combinations and individual treatments of the fungus grown on autoclaved wheat seed, M. javanica eggs (76,000 per plot), and nematicides were applied to specified microplots at the time of transplanting tobacco the first year. Vetch was planted as a winter cover crop, and the fungus and nematicides were applied again the second year to specified plots at transplanting time. The fungus did not control the nematode in either year of these experiments. The average root gall index (0 = no visible galls and 5 = > 100 galls per root system) ranged from 2.7 to 3.9 the first year and from 4.3 to 5.0 the second in nematode-infested plots treated with nematicides. Plants with M. javanica alone or in combination with P. lilacinus had galling indices of 5.0 both years; the latter produced lower yields than all other treatments during both years of the study. Nevertheless, the average soil population densities of P. lilacinus remained high, ranging from 1.2 to 1.3 x 106 propagules/g soil 1 week after the initial inoculation and from 1.6 to 2.3 x 104 propagules/g soil at harvest the second year. At harvest the second year the density of fungal propagules was greatest at the depth of inoculation, 15 cm, and rapidly decreased below this level. PMID- 19290258 TI - Peach Leaf Senescence Delayed by Criconemella xenoplax. AB - Fall annual leaf senescence of peach was delayed in the field and in microplots in the presence of Criconemella xenoplax. Soil from the rhizosphere of orchard trees with greener leaves had ca. 2.5 x more nematodes than soil around trees in a more advanced state of fall senescence. In microplots, monoclonal antibody enzyme immunoassay (EIA) of leaf cytokinins indicated that concentration of zeatin riboside-like substances and chlorophyll content were greater in leaves of trees growing in nematode-infested soil than in trees in uninfested soil. EIA also indicated the presence of substances resembling trans-zeatin, zeatin riboside, dihydrozeatin, and dihydrozeatin riboside-like substances in whole body homogenates of C. xenoplax. Levels of zeatin-like substances were present in the nematode in greater levels than the other related substances. PMID- 19290259 TI - Effect of Simulated Acid Rain on Bursaphelenchus xylophilus Infection of Pine Seedlings. AB - White, Scots, and Austrian 3-year-old pine seedlings were treated with conditions simulating acid rain and inoculated with the white pine specific pathotype of Bursaphelenchus xylophilus, VPSt-1. Oleoresin concentration increased slightly and carbohydrate concentration decreased in all seedlings treated with simulated acid rain (SAR). The changes were significantly increased after inoculation of SAR-treated white and Scots pine seedlings with VPSt-1. Wilting was delayed and nematode reproduction decreased in SAR-treated white pine seedlings inoculated with VPSt-1. SAR-treated Austrian pine seedlings were resistant to VPSt-1, but SAR-treated Scots pine seedlings lost tolerance to VPSt-1 and wilted 50-60 days after inoculation. PMID- 19290260 TI - Reproduction of Meloidogyne javanica on Plant Roots Genetically Transformed by Agrobacterium rhizogenes. AB - Reproduction of Meloidogyne javanica was compared on several Agrobacterium rhizogenes-transformed root cultures under monoxenic conditions. M. javanica reproduced on all transformed roots tested; however, more females and eggs were obtained on potato and South Australian Early Dwarf Red tomato than on bindweed, Tropic tomato, lima bean, or carrot. Roots that grew at moderate rates into the agar and produced many secondary roots supported the highest reproduction. Numbers of females produced in cultures of transformed potato roots increased with increasing nematode inoculum levels, whether inoculum was dispersed eggs or juveniles. Females appeared smaller, produced fewer eggs, and were found in coalesced galls at the higher inoculum levels. The ratio between the final and initial population decreased sharply as the juvenile inoculum increased. The second-stage juvenile was preferred to dispersed eggs or egg masses for inoculation of tissue culture systems because quantity and viability of inoculum were easily assessed. Meloidogyne javanica reared on transformed root cultures were able to complete their life cycles on new transformed root cultures or greenhouse tomato plants. PMID- 19290261 TI - Extremely Sensitive Thermotaxis of the Nematode Meloidogyne incognita. AB - Eggs of the root-knot nematode Meloidogyne incognita were acclimated to 23 C. Newly hatched second-stage juveniles migrated toward higher temperatures when placed in shallow thermal gradients averaging 23 C. The threshold gradient for this response was below 0.001 C/cm, with a best estimate of 4 x 10 C/cm. Calculations of physical limitations on thermotaxis indicate that this sensitivity is well within the limits of what is physically possible. PMID- 19290262 TI - Characterization of Carbohydrates on the Surface of Second-stage Juveniles of Meloidogyne spp. AB - Fluorescent conjugates of the lectins soybean agglutinin (SBA), Concanavalin A (Con A), wheat germ agglutinin (WGA), Lotus tetragonolobus agglutinin (LOT), and Limulus polyphemus agglutinin (LPA) bound primarily to amphidial openings and amphidial secretions of viable, preinfective second-stage juveniles (J2) of Meloidogyne incognita races 1 and 3 (Mil, Mi3) and M. javanica (Mj). No substantial difference in fluorescent lectin binding was observed among the populations examined. Binding of only LOT and LPA were inhibited in the presence of 0.1 M competitive sugar. Structural differences in amphidial carbohydrate complexes among populations of Mi 1, Mi3, and Mj were revealed by glycohydrolase treatment of preinfective J2 and subsequent labeling with fluorescent lectins. A quantitative microfiltration enzyme-linked lectin assay revealed previously undetected differences in lectin binding to nonglycohydrolase-treated J2. Freinfective J2 of Mj bound the greatest amount of SBA, LOT, and WGA, whereas J2 of Mil bound the most LPA. PMID- 19290263 TI - Host Status of Five Weed Species and Their Effects on Pratylenchus zeae Infestation of Maize. AB - The host suitability of five of the most common weed species occurring in maize (Zea mays L.) fields in South Africa to Pratylenchus zeae was tested. Based on the number of nematodes per root unit, mealie crotalaria (Crotalaria sphaerocarpa) was a good host; goose grass (Eleusine indica), common pigweed (Amaranthus hybridus), and thorn apple (Datura stramonium) were moderate hosts; and khaki weed (Tagetes minuta) was a poor host. Only the root residues of khaki weed suppressed the P. zeae infestation of subsequently grown maize. When goose grass, khaki weed, and mealie crotalaria were grown in association with maize in soil infested with P. zeae, goose grass and khaki weed severely suppressed maize root development; this resulted in a low number of nematodes per maize root system and a high number of nematodes per maize root unit. Mealie crotalaria did not restrict maize root growth and did not affect nematode densities per maize root system or maize root unit. Special attention should be given to the control of mealie crotalaria, which is a good host for P. zeae, and goose grass, which, in addition to its ability to compete with maize, is also a suitable host for P. zeae. PMID- 19290264 TI - Standardization of nematicide application rates. PMID- 19290265 TI - Bibliography of Estimated Crop Losses in the United States Due to Plant-parasitic Nematodes. AB - Recent published estimates of crop losses due to plant-parasitic nematodes are summarized by crop and state. Estimates are systematically reported from major producing states for cotton, peanut, tobacco, and soybean. Only scattered reports were available for nematode losses to other field crops or to vegetables and ornamentals. Among the states, North Carolina and Michigan were most consistent in reporting loss estimates from the widest range of crops. PMID- 19290266 TI - Host Suitability of Commercial Corn Hybrids to Meloidogyne arenaria and M. incognita. AB - The host suitability of 64 commercial corn hybrids for a Meloidogyne arenaria race 2 population and a M. incognita race 4 population was determined in greenhouse experiments. 'Northrup King 508' and 'Pioneer Brand 3147' maintained M. arenaria below and at the initial population level, respectively, indicating that these hybrids are relatively poor hosts for this species. RF values (final egg number/initial egg number) of the hybrids for M. arenaria ranged from 0.8 for Northrup King 508 to 42.3 for 'Pioneer Brand XC941'. All hybrids were excellent hosts for M. incognita with RF values ranging from 20.7 for 'Sunbelt 1860' to 49.5 for Pioneer Brand XC941. PMID- 19290267 TI - Nematodes associated with potato in prince edward island and new brunswick. AB - Analysis of soil and root samples collected from potato fields across Prince Edward Island and the Woodstock-Grand Falls region of New Brunswick, Canada showed that Pratylenchus spp. were the dominant plant-parasitic nematodes. Pratylenchus penetrans occurred more often on Prince Edward Island, and P. crenatus predominated in New Brunswick. It was estimated that about 10% of the sites from Prince Edward Island had population levels of Pratylenchus spp. that may have caused economic losses. PMID- 19290268 TI - Geographical Distributions of Rotylenchulus reniformis, Meloidogyne incognita, and Tylenchulus semipenetrans in the Lower Rio Grande Valley as Related to Soil Texture and Land Use. AB - A survey was conducted over a 22-year period to evaluate the influence of soil texture and land use on the geographical distributions of Rotylenchulus reniformis, Meloidogyne incognita, and Tylenchulus semipenetrans in the lower Rio Grande valley. The distributions of R. reniformis and M. incognita were related to soil texture, whereas T. semipenetrans occurred wherever host plants were present regardless of soil texture. The incidence of M. incognita was greatest in elevated sandy loams and moderately well-drained silts of modern flood terraces of the Rio Grande river. Rotylenchulus reniformis occurred predominantly in clay silts and clays of ancient flood terraces. Clay loams and sandy clay loams of the central, irrigated portion of the lower Rio Grande valley appeared favorable for M. incognita and R. reniformis. Differences between the geographical distributions of these two species could not be attributed to host crops. PMID- 19290269 TI - Control of Pratylenchus brachyurus with Selected Nonfumigant Nematicides on a Tolerant and a Sensitive Soybean Cultivar. AB - A field study was conducted to evaluate soybean cultivar sensitivity to Pratylenchus brachyurus and selected nonfumigant nematicides for control of this nematode. 'Essex', a tolerant cultivar, yielded more than 'Forrest' , a sensitive cultivar, in an infested field. Plots treated with aldicarb, carbofuran, and fenamiphos had fewer nematodes 40 days after planting than nontreated plots. Plots planted with Forrest and treated with carbofuran had a greater yield than the untreated controls. PMID- 19290270 TI - Reproduction of Pratylenchus hexincisus and P. scribneri in Corn Inbreds. AB - Population development of lesion nematodes was measured in 17 inbred lines of South Dakota and A619Ht dent corn. In two greenhouse groundbed tests, lines SD 101, SD 102, and SD 103 supported fewer than 1,000 Pratylenchus hexincisus per gram of dry root after 12 weeks. In an irrigated field test, inbred SD101 supported fewer than 1,000 P. scribneri per gram of dry root on each of two sampling dates, whereas line A619Ht supplied high populations of P. scribnerni on both dates. Inbreds SD45, 84742, and 84763 supported high populations of both P. hexincisus and P. scribneri. PMID- 19290271 TI - Evaluation of Soybean Cultivars for Production in Meloidogyne incognita-infested Soil. AB - Significant (P < 0.05) differences among galling and yields of 41 soybean cultivars and breeding lines were found when they were produced at a site infested with Meloidogyne incognita during 3 years of investigation. Over a period of 6 years of testing, 13 cultivars were identified as having a suitably low susceptibility to warrant their production in M. incognita infested soil. PMID- 19290272 TI - Effect of Heterodera glycines on Charcoal Rot Severity in Soybean Cultivars Resistant and Susceptible to Soybean Cyst Nematode. AB - Field experiments were conducted in two soil types in northeastern Kansas to evaluate the influence of Heterodera glycines on the severity of charcoal rot in group III soybean cultivars resistant and susceptible to soybean cyst nematode race 3. Resistant cultivars Asgrow 3307 and Fayette and susceptible cultivars Asgrow 3127, Harper, Pella, Sprite, and Williams 82 were planted in carbofuran treated and nontreated plots. Heterodera glycines and the charcoal rot fungus, Macrophomina phaseolina, were suppressed by carbofuran treatment in loamy sand, but not loam soil, and by nematode-resistant cultivars in both soils. Root densities of the fungus were positively correlated with nematode densities and negatively correlated with seed yield at both locations. Results indicate that H. glycines infection can increase colonization of soybean roots by M. phaseolina which may increase losses due to charcoal rot. PMID- 19290273 TI - Parasitism of Woody Ornamentals by Meloidogyne hapla. AB - Meloidogyne hapla is the dominant root-knot nematode found in Tennessee woody ornamental nurseries. In greenhouse tests, M. hapla produced galls and formed egg masses on roots of Abelia x grandiflora, Comus florida, Hydrangea paniculata grandiflora, Photinia x fraseri, Spiraea x bumalda, Spiraea x vanhouttei, and Viburnum carlesii. Galls on H. grandiflora and V. carlesii were mostly large and fusiform. Galls on C. florida were spherical and usually terminal, whereas those on the other species were minute. Lateral roots grew from galls on all susceptible plants. Two Acer spp., two Buxus spp., three llex spp., five Prunus spp., three Rhododendron spp., Euonymus alata, Ligustrum sinense, Magnolia x soulangiana, Nandina domestica, and nine conifer species were nonhosts or very poor hosts. PMID- 19290274 TI - Response of Cowpea Breeding Lines and Cultivars to Meloidogyne incognita and M. arenaria. AB - Twenty-four cowpea breeding lines and four cultivars were tested for resistance to Meloidogyne incognita in the greenhouse. Gall and egg mass ratings indicated a range of responses from susceptible to highly resistant. Five breeding lines - VS84-2, VS84-8, VS84-12, VS84-14, and VS84-22 - and the cultivar Erectset had gall and egg mass ratings comparable to the M. incognita-resistant cultivar Mississippi Silver. All of these were also resistant to M. arenaria. Significantly fewer M. incognita juveniles were found in roots of resistant than in roots of susceptible plants at 7 and 21 days after inoculation; however, no differences were found at 14 days. PMID- 19290275 TI - Results of annual phytoparasitic nematode surveys of arkansas soybean fields, 1978-1986. AB - Results of surveys from 1978 to 1986 to estimate and identify nematodes in Arkansas soybean fields are presented. The seven most common nematode species in the fields were Heterodera glycines, Quinisulcius acutus, Pratylenchus scribneri, P. alleni, Helicotylenchus pseudorobustus, Tylenchorhynchus ewingi, and Xiphinema americanum. Other nematodes identified from these fields were Pratylenchus brachyurus, P. vulnus, P. zeae, Tylenchorhynchus canalis, T. goffarti, T. martini, Helicotylenchus dihystera, Scutellonema bradys, Xiphinema chambersi, X. rivesi, Hoplolaimus galeatus, H. magnistylus, Paratrichodorus minor (P. christiei), Paratylenchus projectus, P. tenuicaudatus, Criconemella macrodora, C. ornata, and Meiodorus hollisi. PMID- 19290276 TI - Comparison of methyl bromide and other nematicides for control of nematodes in peanut. AB - The efficacy of methyl bromide for control ofMeloidogyne arenaria and to increase yields of 'Florunner' peanut (Arachis hypogaea) was studied in a field at the Wiregrass Substation near Headland, Alabama. Methyl bromide was applied in the row at a depth of 35 cm using a subsoiler-bedder 2 weeks before planting at rates of 0, 34, 50, 67, 101, and 118 kg a.i./ha. Methyl bromide treatments of 67 kg a.i./ha or higher resulted in significant (P = 0.05) yield increases similar to those obtained in the same experiment with at-plant applications of aldicarb (2.2 kg a.i./ha), EDB (1.55 ml a.i./m row), or 1,3-D (5.10 ml a.i./m row). The relation between yield (Y) and methyl bromide rate (x) was described (R(2) = 0.97**) by the exponential function: Y = 2,302.963eb, where b = (- 1.901 - ln x)(2)/169.482. M. arenaria juvenile populations in soil in mid-August were too low to permit establishment of a relation between application rate of methyl bromide and size of the population. PMID- 19290277 TI - Management of Hoplolaimus columbus with Tolerant Soybean and Nematicides. AB - Two experiments, one site per year, were conducted in Scotland County, North Carolina, to determine the usefulness of selected cultivars and nematicides for limiting soybean losses due to Hoplolaimus columbus. Coker 317 was relatively tolerant to this nematode, and Coker 156, Centennial, Dehapine 105, and Gordon were generally intolerant. Most nematicides significantly increased soybean yields, and many gave an economic return. PMID- 19290278 TI - Soybean Response to a Planting-Time Application of Ethylene Dibromide in a Soil Infested with Meloidogyne incognita, M. arenaria, and Heterodera glycines. AB - A field study was conducted to evaluate one susceptible and six nematode resistant soybean cultivars for their effects on seed yield, nematode populations, and response to a fumigant nematicide, ethylene dibromide. The soil was a sandy loam, and the field was infested with a mixture of Meloidogyne incognita, M. arenaria, and Heterodera glycines. Soybean cultivars significantly affected yield and juvenile numbers of Meloidogyne spp. but did not affect the H. glycinesjuvenile population. Fumigation increased yield and numbers of H. glycines juveniles, whereas the numbers of Meloidogyne spp. juveniles were decreased. The interaction between cultivars and fumigation treatment was significant for yield but not for nematode numbers. PMID- 19290279 TI - Effects of Nematicides and Herbicides Alone or Combined on Meloidogyne incognita Egg Hatch and Development. AB - The effects of nematicides carbofuran (C) and fenamiphos (F) and herbicides metribuzin (M) and trifluralin (T), alone and in combination, on hatching, penetration, development, and reproduction of Meloidogyne incognita race 3 were determined under laboratory conditions. To study hatching, entire egg masses were exposed to nematicides (6 mug/ml), herbicides (0.5 mug/ml), and their combinations over a period of 16 days; the hatched juveniles were extracted and counted every 48 hours. Second-stage juveniles that hatched from day 6 to day 8 were used as inoculum to determine the effects of the chemicals on penetration, development, and reproduction of M. incognita on tomato 4, 16, and 32 days after inoculation. F, F + T, and F + M inhibited hatching; whereas, C, T, M, C + T, and C + M did not affect hatching, penetration, development of females, or reproduction. Since so few juveniles hatched from the fenamiphos treatments, we were not able to use them for the postinfection development study. There was no apparent reduction in the effect of the nematicides by the herbicides. PMID- 19290280 TI - Relationship of Yields and Pratylenchus spp. Population Densities in Dryland and Irrigated Corn. AB - Analyses of covariance were used to relate corn yields to population densities of Pratylenchus hexincisus in South Dakota dryland corn or P. scribneri in irrigated corn. The relationship for P. hexincisus was Y = 5,825 - 0.14X - 0.03X' where Y = yield (kg/ha), X = number of P. hexincisus per gram of dry root at midseason and X' = number at harvest. The relationship for P. scribneri was best described by Y = 7,400 - 0.06X' for nematode populations measured at harvest. Yield loss estimates indicated P. hexincisus was more damaging to dryland corn than was P. scribneri to irrigated corn. PMID- 19290281 TI - A Modified Screening Test for Determining Heterodera glycines Resistance in Soybean. AB - A modified version of a standard greenhouse bioassay for determining soybean cyst resistance in soybean plants is described. White plastic laundry tubs served as microplots for rearing large numbers of nematodes in a confined space; up to 3 million eggs of each generation were collected per tub. Before screening, SCN populations were evaluated on susceptible and resistant soybean to characterize female development; these were periodically retested. Screening tests took place in Todd planter flats (120 plants per flat). Test plants were inoculated with 1,200 eggs per plant and evaluated for resistance 33-37 days after inoculation. The plants were pruned at the cotyledonary node which resulted in a greatly reduced root system. Staining the roots in Toluidine Blue created contrast with the white females and facilitated counting. Greenhouse space was conserved, and the labor to set up and maintain the screening test was reduced. PMID- 19290282 TI - Effects of aldicarb on nematodes, early season insect pests, and yield of soybean. AB - The effects of aldicarb on soybean cyst (Heterodera glycines) and root-knot (Meloidogyne incognita and M. arenaria) nematode populations, early season insect pests and soybean (Glycine max) yield were evaluated in five field experiments in northern and southern Alabama. Aldicarb significantly (P = 0.05) reduced nematode populations in only two cases: M. arenaria in Centennial soybean in the Wiregrass site and M. incognita in Bedford soybean in a Tennessee Valley site. No significant difference (P = 0.05) in mean percentage main stem or petiole girdling by threecornered alfalfa hopper (Spissistilus festinus) or in mean number of plants damaged by lesser cornstalk borer (Elasmopalpus lignosellus) occurred among treatments in any experiment. Soybean yields were significantly (P = 0.05) increased in only two cases: in the nematode susceptible Essex and Cobb cultivars planted in the Tennessee Valley and Gulf Coast sites, respectively. Unusually dry 1986 weather conditions may have reduced the activity of aldicarb. PMID- 19290283 TI - Comparison of 'Fayette' Soybean, Aldicarb, and Experimental Nematicides for Management of Heterodera glycines on Soybean. AB - The efficacies of Heterodera glycines-resistant 'Fayene' soybean and aldicarb for managing H. glycines were compared to the experimental nematicides DS-47187 10F, DS-47357 10F, DS-48145 10F, DS-48165 10F, DS-46995 10F and 5G, and DS-38697 5G during 1981-83. Yield of Fayette was greater than yield of the H. glycines susceptible cultivar treated with nematicide in 1981 and 1983. Yield of aldicarb treated soybean was greater than yield of soybean treated with experimental nematicides in 1983. There were no yield differences in 1982. Fewer white females were recovered from Fayette 5 weeks after planting than from soybean treated with nematicides in 1981 and 1982, but not in 1983. Fewer white females were recovered from aldicarb-treated soybean than from experimental nematicide-treated soybean in 1983 but not in 1981 and 1982. In 1983 numbers of first generation white females at 5 weeks and the ratio of those white females to gravid cysts at planting were negatively correlated with soybean yield when soybean was severely damaged by the nematode, but the ratio of final eggs and second-stage juveniles to initial eggs and second-stage juveniles was not correlated with yield. PMID- 19290284 TI - Effect of two nonfumigant nematicides on corn grown in two adjacent fields infested with different nematodes. AB - The organo-phosphate experimental nematicide, O-ethyl S,S-di-sec-butyl phosphorodithioate (FMC 67825), provided yield increases of corn comparable to carbofuran. Both the emulsifiable concentrate and granular formulations of FMC 67825 were equally effective. The evaluations, duplicated in two adjacent fields, clearly demonstrated the importance of the type of plant pathogenic nematodes infesting the sites. Where Belonolaimus longicaudatus occurred, yield increases ranged from 73% to 80%, whereas in the adjacent field (without B. longicaudatus) yields increases ranged from - 14% to 28%. PMID- 19290285 TI - Soybean Response to Ethylene Dibromide in a Soil Infested with Meloidogyne arenaria and Heterodera glycines. AB - One susceptible and six nematode-resistant soybean cultivars were evaluated in the field for their effects on seed yield, nematode populations, and response to the fumigant nematicide, ethylene dibromide. The soil was a loamy sand infested with Meloidogyne arenaria and Heterodera glycines. Cultivars significantly affected yield and numbers of H. glycines but did not affect M. arenaria numbers. Fumigation increased yield and reduced M. arenaria numbers but did not affect numbers of H. glycines. The interaction between cultivars and fumigation was significant for yield but not for nematode numbers. PMID- 19290286 TI - Investigations of the Host Range of the Corn Cyst Nematode, Heterodera zeae, from Maryland. AB - The host range of the corn cyst nematode, Heterodera zeae, recently detected in Maryland, was investigated. A total of 269 plant entries, representing 68 families, 172 genera, and 204 species, was inoculated with cysts or a mixture of eggs and second-stage juveniles of H. zeae. The host range of the Maryland population of H. zeae was limited to plants of the Gramineae and included 11 tribes, 33 genera, 42 species, and 77 entries. All 22 corn (Zea mays) cultivars tested were hosts. Other economic hosts included certain cultivars of barley (Hordeum vulgare), oat (Arena sativa), rice (Oryza sativa), sorghum (Sorghum bicolor), sugar cane (Saccharum interspecific hybrid), and wheat (Triticum aestivum). Fall panicum (Panicum dichotomiflorum), a weed species common to cultivated fields in Maryland, was also a host for H. zeae. Other hosts included meadow foxtail (Alopecurus pratensis), Calamagrostis eipgeios, Job's tears (Coix Lachryma-Jobi), green sprangletop (Leptochloa dubia), witchgrass (Panicum capillare), broomcorn (Panicum miliaceum), fountain grass (Pennisetum rueppeli), reed canary grass (Phalaris arundinacea), common reed (Phragmites australis), eastern gamagrass (Tripsacum dactyloides), corn (Zea mays), and teosinte (Zea mexicana). PMID- 19290287 TI - Effect of combining soil solarization with certain nematicides on target and nontarget organisms and plant growth. AB - Field experiments compared pesticidal and plant growth effects of soil solarization, alone and in combination, with overall applications of several nematicides. Nematodes, including Meloidogyne incognita J2, that were targeted for control were significantly reduced (P < 0.05) by solarization, 1,3 dichloropropene (44 and 132 liter/ha), ethoprop (13.5 kg/ha), metham sodium (64 liter/ha), formaldehyde (111 liter/ha), and by solarization-nematicide combinations. Control of Pythium ultimum also was obtained by all of the treatments; however, none of the chemicals or combinations of chemicals and solarization controlled nematodes or P. ultimum significantly better than solarization alone. Numbers of cotton (Gossypium hirsutum cv. Acala SJ-2) seed applied Trichoderma viride and Bacillus subtilis which colonized the plant rhizosphere were not affected. Yield of carrot and survival of cotton seedlings was sometimes increased by solarization and (or) chemical treatments. No significant phytotoxicity from soil treatments was found on cotton or carrot. PMID- 19290288 TI - Comparative Resistance of Selected Acala 1517 Cotton Cultivars to Meloidogyne incognita Race 3. AB - Little information is available regarding the levels of Meloidogyne incognita race 3 resistance in Acala 1517 cotton cultivars compared with cultivars grown outside the southwestern United States. Levels of M. incognita egg production were compared among commercial Acala cultivars 1517-E2, 1517-SR1, 1517-75, 1517 77BR, and SJ-5, resistant and susceptible standards Auburn 634 and M-8 and breeding lines Acala 5701-W and N6072 grown for 45 days in the greenhouse. The Acala 1517 cultivars all performed similarly, demonstrating moderate nematode resistance. Egg production on the 1517 cultivars was less than on SJ-5 and less than one-fifth that on M-8. Auburn 634 was most resistant, followed by N6072. Total egg production per plant and egg production per gram dry root were not correlated but proved to be reliable indicators of relative resistance to M. incognita. PMID- 19290289 TI - Nematode response to cool season annual graminaceous species and cultivars. AB - The response of 29 rye, oat, triticale, and wheat cultivars to selected nematode species was determined in the greenhouse. Variability in nematode root galling and final nematode population densities in root and soil in response to cool season annual graminaceous crops occurred for spiral (Helicotylenchus dihystera), stubby root (Paratrichodorus minor), and root-knot (Meloidogyne incognita) nematodes. Although none of the graminaceous crops supported M. incognita at levels as high as the susceptible 'Davis' soybean control, sufficient variation existed among these to warrant field scale studies. PMID- 19290290 TI - Efficacy of Using Split and Postplant Applications of Aldicarb for Control of Helerodera schachtii on Sugarbeet. AB - Soil temperature at planting and initial population densities (Pi) significantly affected (P < 0.05) the chemical control of Heterodera schachtii on sugarbeet, Beta vulgaris. The fumigant 1,3-dichloropropene (1,3-D) at 9.4 g/m of row effectively controlled H. schachtii, resulting in increased sugarbeet yields over the nontreated control treatment at soil temperatures at planting of 8, 12, and 16 C and Pi of 4.7, 10.4, and 18.3 eggs/cm(3) at planting. A split application of aldicarb, 1.3 g/m of row at planting (AP) and 28 days later (PP), and a single at planting (AP) treatment of 2.1 g/m of row were less effective in controlling H. schachtii than 1,3-D at the three soil temperatures and Pi. The sugarbeet yield from the AP plus PP treatment, however, was greater than the sugarbeet yield from the AP treatment and was equivalent to the yield from the 1,3-D treatment at temperatures of 8 and 12 C and a Pi of 4.7 eggs/cm(3). Sugarbeet yield from the AP treatment was significantly (P < 0.05) greater than the AP plus PP application at a Pi of 18.3 at a planting temperature of 8 C and Pi of 10.4 and 18.3 eggs/cm(3) at soil planting temperatures of 12 and 16 C. Postplant application of 2.1 g/m of aldicarb, applied 28 days after planting, significantly increased sugarbeet yields at all soil temperatures at the lower Pi levels. PMID- 19290291 TI - Control of Hoplolaimus columbus on Late-planted Soybean with Aldicarb. AB - Efficacy of three rates of aldicarb for control of Hoplolaimus columbus was evaluated on highly tolerant ('Foster'), moderately tolerant ('Coker 368'), and highly susceptible ('Braxton') soybean cultivars planted on 28 June 1985. Average yields of the nontreated cultivars were 557 kg/ha for Braxton, 1,309 kg/ha for Coker 368, and 1,682 kg/ha for Foster. Yield differences were significant. Aldicarb increased yields of all three cultivars, with the most tolerant cultivar responding to all aldicarb rates and the most susceptible cultivar responding only to the low rate of aldicarb; there was no significant response to the higher rates. H. columbus population densities in the soil 6 weeks after planting were not affected by either aldicarb treatment or cultivar, whereas the populations from roots were reduced in all cultivars by aldicarb. No relationship between tolerant soybean cultivars and population densities of H. columbus was observed. PMID- 19290292 TI - First Report of a Tylenchulus sp. on Peach in Alabama, Arkansas, Georgia, and South Carolina. AB - A Tylenchulus sp. found in a Georgia peach orchard parasitized peach roots in the greenhouse. Citrus roots were not parasitized, indicating that the nematode was not the citrus nematode. Morphologically similar populations were found in one peach orchard in Alabama, two orchards in Arkansas, and one in South Carolina. Males were present in the Alabama populations and one of the Arkansas populations. A population was also found in an area of mixed hardwood and pine in Arkansas. The populations are morphologically different from the citrus nematode Tylenchulus semipenetrans. PMID- 19290293 TI - Application of nematicides via drip irrigation. PMID- 19290294 TI - Screening of a Granular Chelate of Metham-Zinc for Nematicidal Activity Using Citrus and Root-knot Nematodes. AB - A granular formulation of a chelate of metham-zinc (CMZ) which liberates the biocidal methyl isothiocyanate was tested for nematicidal activity on Tylenchulus semipenetrans in a jar soil screening and on Meloidogyne javanica (greenhouse test) and M. incognita (field test) infecting tomato. Comparisons were made with 1,3-D in the jar and pot experiments. The CMZ caused only 3.9% mortality of citrus nematode juveniles at 1.0 mug a.i./g soil, but 95.4% mortality at 10.0 mug a.i./g and 100.0% at 100.0 mug a.i./g. CMZ at 10.0 and 100.0 mug a.i./g significantly reduced tomato root infections by M. javanica in the pot test relative to the untreated control. In the field test, CMZ (11.5 g a.i./m(2) calibration rate) reduced M. incognita populations in the zone of incorporation but not below it, thus failing to provide season-long control for tomato. This material has good nematicidal activity at 10 mug a.i./g or more, but its effectiveness in the field may be limited by its lack of movement. PMID- 19290295 TI - Nematode population densities and yield of sweet potato and onion as affected by nematicides and time of application. AB - Nematode population densities and yield of sweet potato and onion as affected by nematicides and time of application were determined in a 3-year test. Population densities of Meloidogyne incognita race 1 in untreated plots of sweet potato increased each year, but Helicotylenchus dihystera and Criconemella ornata did not. Ethoprop (6.8 kg a.i./ha) incorporated broadcast in the top 15-cm soil layer each spring before planting sweet potato reduced population densities of nematodes in the soil and increased marketable yield in 1982, but not in 1983 and 1984. When DD, fenamiphos, and aldicarb were applied just before planting either sweet potato or onion, nematode population densities at harvest were lower in treated than in untreated plots. No additional benefits resulted when the nematicides were applied immediately before planting both sweet potato and onion. Correlation coefficients (P sigma*(C-X) excitation, which may as well be localized on the substituents in this series. Therefore, the observed property cannot be strictly attached to the so-called reaction center. The behavior of substituent constants in the description of core-hole processes in aliphatic compounds is also checked on C1s ionization potentials for the extended series of substituted methanes. The role of the inductive effect for both resonant Auger kinetic energies (E(k)) and C1s binding energies (E) is very important. C(CH(3))1s -->sigma*(C-X) excitation energies do not show any significant correlations with inductive substituent constants. On the contrary, Taft's inductive parameters alone give good correlations for E(k). A simple model with 'Z + 1' approximation is developed to describe E(k) and explain the observed correlations: [formula: see text] where E(b)(v(Z)) is the valence ionization potential for CH(3)X and E(ea(Z+1)(+)) is the electron affinity for [NH(3)X](+). PMID- 19290330 TI - Cyanide orientational ordering and copper electric field gradients in CuCN.N2H4. AB - The measurement of residual dipolar coupling in (13)C and (15)N MAS spectra of CuCN.N(2)H(4) allows for the extraction of (63/65)Cu quadrupole coupling constants (C(Q)((63)Cu) = 26 +/- 4 MHz). Ab initio calculations were employed to determine electric field gradient and chemical shielding tensor orientations, which proved essential for the reliable analyses of residual dipolar coupled (13)C and (15)N MAS spectra. The (1)J((63)Cu,(13)C) and (1)J((63)Cu,(15)N) couplings (590 and -120 Hz, respectively) indicate that the cyanide ligands are static and their magnitudes reflect the deviation of the C-Cu-N angle when compared with other copper cyanides. The observed (13)C and (15)N spectra are most compatible with a structural model wherein the cyanides are fully orientationally ordered: [-Cu-C-N-Cu-C-N-]. PMID- 19290331 TI - Role of structures with penta- and hexacoordinate silicon in the nucleophile catalyzed hydrolysis of tetramethoxysilane. AB - The mechanism of the base catalyzed hydrolysis of tetramethoxysilane (TMOS), proposed earlier on the basis of experimental data, is assessed by theoretical methods, i.e. MP2 and B3LYP with 6-31G(d) and Dunning correlation-consistent basis sets. Models considered involve one and two hydrolyzing water molecules attacking (MeO)(4)Si in the frontside position with a nucleophile (NH(3) and OH( )) in the backside position. This approach allowed us to simulate uniformly the catalytic action of weak and strong bases. It was shown that the presence of a base in the backside position considerably lowers the activation barrier for hydrolysis. The inclusion of the additional water molecule which results in a substantial lowering of the barrier for uncatalyzed hydrolysis does not change noticeably the catalytic effect. In both one- and two-water molecules models the structure of the transition state in the presence of a nucleophile becomes nearly octahedral in which hexacoordinate silicon and four oxygens of the TMOS moiety have a planar arrangement. PMID- 19290332 TI - Interfacial supramolecular cyclodextrin-fullerene assemblies: host reorientation and guest stabilization. AB - Monolayers of mono and bis-pyridine-substituted beta- and gamma-cyclodextrins have been self-assembled onto polycrystalline gold electrodes and electrochemically characterised. Surface enhanced Raman spectroscopy (SERS) has been used to probe the time dependent reorientation of the CDs and to probe the effect of co-adsorbing alkane thiols. Significantly, we observe novel SERS signatures for carbohydrate centred between 2100 and 2300 cm(-1) which is sensitive to the orientation of the CD moiety on the gold surface. These modes are thought to arise from direct interactions between the CD pyranose ring and the gold substrate and are useful markers for assigning orientation of CD within a film. SERS confirmed, as was intimated from a previous electrochemical study, that significant reorientation of the CD groups within the film occurs on annealing and this orientation can be reversed by backfilling with alkane thiol. C(60), in its unmodified state, was then immobilised through host-guest assembly into the backfilled CD monolayers to produce a 2 : 1 CD : C(60) trilayer. These CD capped fullerenes are protected from photopolymerisation and exhibit well defined redox processes. PMID- 19290333 TI - Where does acid hydrolysis take place? AB - We present the results of computations on the dissociation of HCl and HNO(3) at the air-water interface. Molecular dynamics simulations of the acid molecule and 200 water molecules were propagated for several nanoseconds, and the resulting structures were used as input to QM/MM geometry optimization runs. Approximately 20-30 water molecules were included along with the acid in the QM portion of the calculation, which was carried out at the B3LYP/6-31+G(d) level. Whereas dissociation to ions is always spontaneous in the bulk, we find that acid molecules confined to the water surface dissociate only with the participation of two additional water molecules, forming a "critical cluster" about the solute. Thus acid dissociation may occur in the near-surface zone at a dynamic air-water interface, in agreement with our earlier experimental conclusions [Clifford et al., Phys. Chem. Chem. Phys.. 2007, 9, 1362]. PMID- 19290334 TI - Investigation of the Soret effect in aqueous and non-aqueous mixtures by the thermal lens technique. AB - In the present work we investigate the thermal diffusion behavior of three different binary mixtures with a thermal lens (TL) setup. In the setup used in this study we avoid the addition of a dye for systems, such as aqueous mixtures, with a weak absorption band at a wavelength of 980 nm. In some aqueous systems with a complex phase behavior the addition of dye significantly affects the apparent measured thermal diffusion properties. The studied systems are dimethylsulfoxide (DMSO) in water, the ionic liquid 1-ethyl-3-methylimidazolium ethylsulfate (EMIES) in butanol and a non-ionic surfactant hexaethylene glycol monododecyl ether (C(12)E(6)) in water. The Soret coefficients of the selected systems cover a range of two orders of magnitude. For DMSO in water with a very low Soret coefficient of the order of S(T) approximately 10(-3) K(-1) we find for a low DMSO content (c = 0.33) a reasonable agreement with previous measurements, while the weak thermal lens signal for the DMSO-rich mixture (c = 0.87) leads to 20% too large Soret coefficients with an uncertainty of more than 30%. Secondly we studied a liquid salt 1-ethyl-3-methylimidazolium ethylsulfate (EMIES) in butanol with a roughly ten times higher Soret coefficient of S(T) approximately 10(-2) K(-1). For this system we performed additional measurements with another experimental technique, the classical thermal diffusion forced Rayleigh scattering (TDFRS), which requires the addition of a small amount of dye to increase the absorption. In the entire investigated concentration range the results obtained with the TL and classical TDFRS technique agree within the error bars. As a third system we studied a non-ionic surfactant hexaethylene glycol monododecyl ether (C(12)E(6)) in water with a Soret coefficient of the order of S(T) approximately 10(-1) K(-1). For this system we find good agreement with previous measurements. We conclude that the TL technique is a reliable method for systems with a strong optical contrast and fairly large Soret coefficient of the order of S(T) approximately 10(-2) K(-1). PMID- 19290335 TI - The role of acidic sites and the catalytic reaction pathways on the Rh/ZrO2 catalysts for ethanol steam reforming. AB - Rh catalysts supported on ZrO(2)-based oxides were studied for ethanol steam reforming (SR) reaction. Pure ZrO(2) as the support resulted in higher H(2) production yield compared to the ZrO(2) oxide decorated with CeO(2), Al(2)O(3), La(2)O(3) or Li(2)O at the reaction temperature of 300 degrees C. Above 450 degrees C, all the catalysts exhibited similar catalytic activity. However, at low reaction temperatures (below 400 degrees C), a significant enhancement in the catalytic activity, selectivity and stability was achieved by replacing the ZrO(2) support prepared by a precipitation method (ZrO(2)-CP) with that prepared by a hydrothermal method (ZrO(2)-HT). A deactivation was observed during the EtOH SR reaction at 300 degrees C on the two catalysts of Rh/ZrO(2)-CP and Rh/ZrO(2) HT. NH(3)-TPD experiments confirmed that the ZrO(2)-HT support had two types of acidic sites while the ZrO(2)-CP support had only one type of weak acidic sites. DRIFTS studies showed that the absorption of EtOH molecules was strong on the Rh/ZrO(2)-HT catalyst and a number of C(2) oxygenates were accumulated on the catalyst surface. Meanwhile, the EtOH absorption on the Rh/ZrO(2)-CP catalyst was weak and the accumulation of CO, carbonate and CH(x) was observed. It is concluded that the relatively strong Lewis acidic sites in the Rh/ZrO(2)-HT catalyst is responsible for the strong absorption of EtOH molecules, and the subsequent C-H breakage step (formation of acetaldehyde or called as dehydrogenation reaction) is a fast reaction on it; on the Rh/ZrO(2)-CP catalyst, the EtOH adsorption was weak and the C-C breakage was the dominating reaction which led to the accumulation of surface CO, CH(x) and CO(2) species. Therefore, it is believed that, in order to promote the absorption of EtOH molecules and to reduce the formation of metastable carbonaceous species (C(2) oxygenates) during the reaction, the catalyst should be enhanced both with Lewis acidity and with C C bond breakage function. Also, it was found that the Rh particle size and distribution, as well as the surface area of the catalyst, were not important factors in determining the catalytic performance. PMID- 19290336 TI - Quantum dynamics of light-driven chiral molecular motors. AB - The results of theoretical studies on quantum dynamics of light-driven molecular motors with internal rotation are presented. Characteristic features of chiral motors driven by a non-helical, linearly polarized electric field of light are explained on the basis of symmetry argument. The rotational potential of the chiral motor is characterized by a ratchet form. The asymmetric potential determines the directional motion: the rotational direction is toward the gentle slope of the asymmetric potential. This direction is called the intuitive direction. To confirm the unidirectional rotational motion, results of quantum dynamical calculations of randomly-oriented molecular motors are presented. A theoretical design of the smallest light-driven molecular machine is presented. The smallest chiral molecular machine has an optically driven engine and a running propeller on its body. The mechanisms of transmission of driving forces from the engine to the propeller are elucidated by using a quantum dynamical treatment. The results provide a principle for control of optically-driven molecular bevel gears. Temperature effects are discussed using the density operator formalism. An effective method for ultrafast control of rotational motions in any desired direction is presented with the help of a quantum control theory. In this method, visible or UV light pulses are applied to drive the motor via an electronic excited state. A method for driving a large molecular motor consisting of an aromatic hydrocarbon is presented. The molecular motor is operated by interactions between the induced dipole of the molecular motor and the electric field of light pulses. PMID- 19290337 TI - Selective observation of the two oxygen atoms at different sites in the carboxyl group (-COOH) of liquid acetic acid. AB - A study of the excitation energy dependence of resonant X-ray emission spectra across the O1s absorption edge in combination with X-ray absorption spectroscopy and density functional theory reveals the site-specific valence electronic structure of liquid acetic acid under ambient conditions. PMID- 19290338 TI - Thermal stability of interaction between the CO molecules and the Al doped graphene. AB - The thermal stability of interaction between CO molecules and Al doped graphene is studied using ab initio molecular dynamics calculations to reveal the adsorption/desorption behavior of the system. With these results, an adsorption desorption phase diagram was established with atomic thermodynamics. The temperature (T) dependent desorption time tau(T) was determined with a thermal desorption method. The results show that the optimal desorption temperature is 400 K. The effect of T on atomic structure parameters and electrical properties were analyzed. It shows that the maximum variation of electrical conductivity induced by the adsorption occurs at 400 K, indicating that the best sensing performance of the devices should be at this particular temperature. PMID- 19290339 TI - Photophysics of phenalenone: quantum-mechanical investigation of singlet-triplet intersystem crossing. AB - We have examined the electronic and molecular structure of 1H-phenalen-1-one (phenalenone) in the electronic ground state and in the lowest excited states, as well as intersystem crossing. The electronic structure was calculated using a combination of density functional theory and multi-reference configuration interaction. Intersystem crossing rates were determined using Fermi's golden rule and taking direct and vibronic spin-orbit coupling into account. The required spin-orbit matrix elements were obtained applying a non-empirical spin-orbit mean field approximation. Our calculated electronic energies are in good agreement with experimental data. We find the lowest excited singlet states to be of the npi* (S1) and pipi* (S2) type. Energetically accessible from S1 are two triplet states of the pipi* (T1) and npi* (T2) type, the latter being nearly degenerate to S1. This ordering of states is retained when the molecular structure in the electronically excited states is relaxed. We expect very efficient intersystem crossing between S1 and T1. Our calculated intersystem crossing rate is approximately 2 x 10(10) s(-1), which is in excellent agreement with the experimental value of 3.45 x 10(10) s(-1). Our estimated phosphorescence and fluorescence rates are many orders of magnitude smaller. Our results are in agreement with the experimentally observed behavior of phenalenone, including the high efficiency of 1O2 production. PMID- 19290340 TI - Infrared and microwaves at 5.8 GHz in a catalytic reactor. AB - An improved micro-reactor cell for IR spectroscopic studies of heterogeneous catalysis was built around a 5.8 GHz microwave cavity. The reactor can operate at 20 bars and with conventional heating up to 720 K, with reactant gas flows velocities (GHSV) from 25,000 to 50,000 h(-1). The temperature of the sample under microwave irradiation was measured by time resolved IR emission spectroscopy. The first experiment performed was the IR monitoring of the desorption of carbonates induced by irradiating an alumina sample by microwaves at 5.8 GHz. PMID- 19290341 TI - Protonation of water clusters in the cavities of acidic zeolites: (H2O)n.H chabazite, n = 1-4. AB - Proton forms of zeolite chabazite (H-SSZ-13) loaded with 1 to 4 water molecules per acid site are examined by density functional theory with periodic boundary conditions. Equilibrium structures are determined by localizing minima on the potential energy surface and harmonic vibrational frequencies are calculated. Average structures, proton dynamics and anharmonic spectra at finite temperature (350 K) are determined by molecular dynamics (MD). The protonation state is found to depend on the number of water molecules per acid site (loading) following the trend of increasing proton affinity with increasing cluster size. Single water molecules are not protonated, the protonated water dimer is the most stable equilibrium structure with the PBE functional, but not with BLYP. MD shows that even with PBE, the protonated water dimer is not stable at finite temperature. The protonated water trimer may be formed as a short-lived species, but the protonated water tetramer is the smallest stable protonated cluster. For the same global loading (2 : 1), a heterogeneous distribution of adsorbed water molecules over the cells is more stable than a homogeneous one (1 : 1/3 : 1 vs. 2 : 1/2 : 1 for a double cell), i.e. non-protonated and protonated water clusters may exist simultaneously in polyhydrated H-SSZ13. Adsorption energies (0 K) per water molecule decrease from 71 to 51 kJ mol(-1) for n = 1 to n = 4. PMID- 19290342 TI - Solar-induced self-assembly of TiO2-beta-cyclodextrin-MWCNT composite wires. AB - Wire-like TiO2-beta-cyclodextrin-MWCNT composites have been successfully fabricated through a solar-induced self-assembly process. The wires were stable for more than several months both in water and under air in dry conditions. The composites were characterized in detail by scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectra, Raman spectra and Raman mapping. The results revealed that the wires with a mean diameter of 1 microm were uniform. MWCNT and beta-cyclodextrin can interact with each other, which was confirmed in our previous research, meanwhile, monodisperse TiO2 nanoparticles can also associate with beta-cyclodextrin. Thus, beta cyclodextrin molecules acted as linkers between MWCNT and monodisperse TiO2 nanoparticles. A reasonable mechanism was also proposed to explain the formation of this wire. Furthermore, the dye-sensitized solar cells were assembled with a sandwich-structured electrode containing the wires and they exhibited an enhanced power conversion efficiency. This may be attributable to the compact composite structure of the wires and the unique electronic properties of MWCNT, which favor the transportation of photogenerated electrons. PMID- 19290343 TI - Influence of the geometry of a hydrogen bond on conformational stability: a theoretical and experimental study of ethyl carbamate. AB - Spectra of ethyl carbamate (urethane) in the gas phase have been recorded in the microwave (4-20 GHz), millimeter-wave (49-118 GHz and 150-235 GHz) and mid infrared (1000-1900 cm(-1)) regions. At the same time, high level ab initio calculations have been performed in order to both predict the experimental results and help in understanding the physical properties of the system. An extensive set of spectroscopic constants for the two most stable conformers in the gas phase, that might be useful for astrophysical databases, has been derived from the observed signals. The most stable conformer has been unambiguously identified. Then, the influence of a weak intramolecular hydrogen bond on the conformational stability has been discussed on the basis of theoretical and experimental results. PMID- 19290344 TI - Reflectance spectroscopy in analysis of UO2 scale: derivation of a kinetic model of uranium oxidation. AB - In this study, we analyzed the development of a compact oxide scale built in course of Uranium surface oxidation. The process was monitored by an in-situ acquisition of the reflectance interference peaks in the NIR-MIR. Dielectric properties of the growing oxide scale were derived in accord to the oscillator model. We used effective media approach to simulate heterogeneous dielectric content in the oxide-metal interface. Following dielectric parameterization, structural properties (e.g., scale thickness) of the proposed multi-scale scheme were calculated. As scale's growth process quantified, a valid kinetic model was proposed. Analysis showed that oxidation dynamics is governed by a multi parabolic, true diffusion-limited mechanism of activation energy conveniently equaling the known anion diffusion enthalpy of 26 kcal/mol. The applied kinetic model suggested a setup of two consecutive oxide scales, characterized by differing anion diffusion rates. Though mathematical formalism presented a similar to the paralinear, time-dependent solution, here, in contrast to the classic paralinear assumption, both scales consisted of a compact, diffusion limited oxide barriers. As a result, the difference in anion flow across the outer and inner scale barriers assigned the overall, pseudo-linear rate constant kl, of a negative (in contrast to the paralinear approach) value. Next, Uranium oxidation has been studied in the post-elastic domain. Markedly, upon breakaway of the compact oxide scale, classic paralinear behavior was reestablished for scale thickness of > or = 0.5 microm. PMID- 19290345 TI - Photodamage to isolated mononucleotides--photodissociation spectra and fragment channels. AB - UV excitation of isolated singly-charged deprotonated mononucleotide anions in the gas phase can lead to their dissociation. We present mass spectrometry results, photodepletion and photofragment action spectra on the UV photodissociation of deprotonated 2'-deoxyribonucleobase-5'-monophosphates with adenine, cytosine, guanine and thymine as nucleobases. We observe the same anionic fragments as in earlier experiments on collision-induced dissociation, although their relative intensities are quite different, especially with respect to the abundance of the deprotonated base anions. The fragment channels correspond to loss of genetic information by cleavage of the CN glycosidic bond and to strand breaking by severing the phosphate-sugar link. We compare the photodissociation spectra with UV absorption spectra of aqueous solutions of the same species and discuss the photodissociation behavior in the context of possible mechanisms and ergodic versus non-ergodic fragmentation. PMID- 19290346 TI - A comparison of the quantum state-specific efficiency of N + N2 reaction computed on different potential energy surfaces. AB - State-to-state exact quantum probabilities of the N + N2 exchange reaction have been calculated on the recently proposed L4 potential energy surface fitted to high level ab initio points using full-dimensional time-independent quantum techniques. Thermal rate coefficient values calculated on L4 were found not to differ from those calculated on a previously proposed potential energy surface. On the contrary, state-specific reaction probabilities calculated on the two surfaces are shown to differ significantly. Arguments for attributing the difference to specific features of the considered potential energy surfaces are provided. PMID- 19290347 TI - Charge delocalization dynamics of ammonia in different hydrogen bonding environments: free clusters and in liquid water solution. AB - Valence and core level photoelectron spectra and Auger electron spectra of ammonia in pure clusters have been measured. The Auger electron spectra of gas phase ammonia, pure ammonia clusters and ammonia in aqueous solution are compared and interpreted via ab initio calculations of the Auger spectrum of the ammonia monomer and dimer. The calculations reveal that the final two-hole valence states can be delocalized over both ammonia molecules. Features at energies pertaining to delocalized states involving one, or more, hydrogen bonding orbitals can be found in both the ammonia cluster Auger electron spectrum and in that of the liquid solvated molecule. The lower Coulombic repulsion between two delocalized valence final state holes gives higher kinetic energy of the Auger electrons which is also observed in the spectra. This decay path--specific to the condensed phase--is responsible for more than 5% of the total cluster Auger intensity. Moreover, this interpretation is also applicable to the solid phase since the same features have been observed, but not assigned, in the Auger spectrum of solid ammonia. PMID- 19290348 TI - Surface states- and field-effects at p- and n-doped GaAs(111)A/solution interface. AB - We have investigated the behaviour of n- and p-doped GaAs(111)A electrodes in sulfuric acid solution by electrochemical impedance spectroscopy (EIS) and second harmonic generation (SHG) measurements. The potential dependence of the SHG response was found to be closely related to the changes in the surface state population, as revealed by analysis of the impedance data. The nature of the majority of carriers turned out to be a key factor in shaping the surface state- and field-effect on the second harmonic generation process. PMID- 19290349 TI - Temperature-dependence of hydrogen oxidation reaction rates and CO-tolerance at carbon-supported Pt, Pt-Co, and Pt-Ru catalysts. AB - The temperature-dependence of the hydrogen oxidation reaction (HOR) rate was examined at commercial Pt, Pt3Co, PtRu, and PtRu(1.5) nano-sized catalysts (diameter, d = ca. 3 nm) supported on carbon black in 0.1 M HClO4 solution in the presence and absence of carbon monoxide by use of a channel flow electrode at temperatures from 30 to 90 degrees C. It was found that the values of the apparent rate constant k(app) (per real Pt active surface area) for the HOR at these supported catalysts agreed beautifully with those of the corresponding bulk electrodes in the whole temperature range. The dependence of the kinetically controlled current density (jk) on CO coverage at each supported catalyst was also identical to that of the bulk. Hence, no particle size effect was observed on the HOR activity and the CO tolerance, at least, was brought down to d = 3 nm. PMID- 19290350 TI - Dielectric properties of differently flexible polyions: a scaling approach. AB - The dielectric relaxations associated with counterion polarization along some typical polyion lengths have been measured in an extended frequency range (from 10 kHz to 2 GHz) for four different polyelectrolyte solutions, differing for the polyion molecular weight and the backbone stiffness. Here, we deal with the so called intermediate dielectric relaxation, falling between the polarization process concerning the whole polyion chain and the polarization process associated with the field-induced re-orientation of the water molecule dipoles. These observed intermediate relaxations have been characterized by means of two parameters, i.e., the dielectric strength Deltaepsilon and the relaxation frequency nu0, and their dependence on the polyion concentration has been described according to the scaling model of a polyelectrolyte solution. These dependencies follow the expected exponents of the scaling laws, both for the dilute and semidilute (unentangled and entangled) regimes. The different concentration regimes evidenced from dielectric relaxation measurements are in very good agreement with the ones determined by means of the zero-shear viscosity measurements. Our results confirm that the intermediate dielectric relaxation in polyelectrolyte solutions should be attributed to counterion fluctuations along some segments (e.g. the subunits of the Mandel model) of the polyion chain, independently of its overall stiffness. This counterion polarization effect is rather confined to the local structure of the polyion chain instead of the whole chain and it is largely independent of the polyion conformation. PMID- 19290351 TI - Self-assembly of a dithiocarbamate calix[4]arene on Ag nanoparticles and its application in the fabrication of surface-enhanced Raman scattering based nanosensors. AB - The absorption and self-assembly of a dithiocarbamate calix[4]arene derivative (DTCX) on Ag nanoparticles (NPs) was characterized in this work by surface enhanced Raman scattering (SERS). This study was carried out on Ag NPs prepared by chemical reduction of silver nitrate with two different reducing agents: sodium citrate or hydroxylamine hydrochloride. SERS was able to discriminate between the different conformations and interaction geometries adopted by DTCX when adsorbed and self-assembled on Ag NPs at different surface coverings. The identification of structural marker bands was crucial in this study. Furthermore, the structure adopted by DTCX is important to determine the activity of this calixarene as a molecular host to detect pollutants, such as pyrene (PYR), since an excessively open or closed cavity is not efficient regarding the analyte detection. PMID- 19290352 TI - Adsorption of oxalate on anatase (100) and rutile (110) surfaces in aqueous systems: experimental results vs. theoretical predictions. AB - A combined experimental and theoretical study of the adsorption of oxalic acid from the aqueous phase at the surface of anatase nanoparticles has been performed. The interfaces were investigated by ATR-FTIR measurements and quantum chemical calculations using the semiempirical method MSINDO. The vibration spectra of the most stable surface complexes have been calculated and used for the interpretation of experimental results. The theoretical studies have been done using the anatase (100) surface to model the adsorption of oxalic acid and water. The effect of interaction of water and oxalic acid on the adsorption mechanism and the vibration spectra was taken into account in the theoretical models. Inclusion of solvation effects was found crucial to determine the type of denticity and structure of adsorbed complexes. By comparison of experimental data and theoretical calculations the most likely surface species and the effects of hydration in their relative stabilities were determined. The present results are compared to previous studies preformed also by combination of experimental and theoretical calculations of analogous systems using nanoparticulate rutile [C. B. Mendive et al., Phys. Chem. Chem. Phys., 2008, 10, 1960, ref. 1]. Differences between surface complexes on anatase and rutile lie mainly on the denticity type. Whilst in the case of rutile the most stable species consist of two bidentate surface complexes followed in third place by a monodentated form, anatase allows the formation of four species in which the stability order is reversed with respect to the denticity type. In the case of anatase, the main contributors to the surface speciation are two monodentate species differing in the position of the H atom within the molecule (being more stable when it is placed in the O-C-O moiety not bound to the surface); and two bidentate species, one deprotonated and one monoprotonated, in which the C-C bond was parallel or perpendicular to the TiO2 surface, respectively. PMID- 19290353 TI - Resonance Raman study of the superoxide reductase from Archaeoglobus fulgidus, E12 mutants and a 'natural variant'. AB - The resonance Raman (RR) spectra of the oxidized wild-type Archaeoglobus fuglidus 1Fe-Superoxide reductase (SOR), E12V and E12Q mutants were studied at different pH conditions upon excitation in resonance with the pH-dependent charge transfer transition to the ferric iron. The wild-type SOR from Nanoarchaeum equitans that lacks the highly conserved glutamate residue was investigated as a 'natural variant'. No substantial differences were observed in the RR spectra of the active sites of the A. fulgidus proteins. Based on the component analysis in the metal-ligand stretching region the modes involving the Fe-S(Cys) stretching coordinates have been identified. The frequencies of these modes reflect the electronic properties of the Fe-S bond which are related to the catalytic activity of SORs, including reduction of superoxide and product dissociation. Moreover, hydroxide binding to the E12 mutant proteins was demonstrated at high pH. It was further observed that the ferric active site of all three SORs from A. fulgidus senses the presence of phosphate, which possibly replaces the hydroxide at high pH. PMID- 19290354 TI - Anthraquinone catalysis in the glucose-driven reduction of indigo to leuco indigo. AB - Anthraquinone immobilised onto the surface of indigo microcrystals enhances the reductive dissolution of indigo to leuco-indigo. Indigo reduction is driven by glucose in aqueous NaOH and a vibrating gold disc electrode is employed to monitor the increasing leuco-indigo concentration with time. Anthraquinone introduces a strong catalytic effect which is explained by invoking a molecular "wedge effect" during co-intercalation of Na+ and anthraquinone into the layered indigo crystal structure. The glucose-driven indigo reduction, which is ineffective in 0.1 M NaOH at 65 degrees C, becomes facile and goes to completion in the presence of anthraquinone catalyst. Electron microscopy of indigo crystals before and after reductive dissolution confirms a delamination mechanism initiated at the edges of the plate-like indigo crystals. Catalysis occurs when the anthraquinone-indigo mixture reaches a molar ratio of 1 : 400 (at 65 degrees C; corresponding to 3 microM anthraquinone) with excess of anthraquinone having virtually no effect. A strong temperature effect (with a composite EA approximately 120 kJ mol(-1)) is observed for the reductive dissolution in the presence of anthraquinone. The molar ratio and temperature effects are both consistent with the heterogeneous nature of the anthraquinone catalysis in the aqueous reaction mixture. PMID- 19290355 TI - Probing local structures of siliceous zeolite frameworks by solid-state NMR and first-principles calculations of 29Si-O-29Si scalar couplings. AB - Subtle structural details of siliceous zeolites are probed by using two-bond scalar (J) coupling constants to characterize covalently bonded 29Si-O-29Si site pairs and local framework order. Solid-state two-dimensional (2D) 29Si{29Si} NMR measurements and first-principles calculations of 2J(29Si-O-29Si) couplings shed insights on both the local structures of siliceous zeolites Sigma-2 and ZSM-12, as well as the sensitivity of J couplings for detailed characterization analyses. DFT calculations on a model linear silicate dimer show that 2J(Si-O-Si) couplings have complicated multiple angular dependencies that make semi-empirical treatments impractical, but which are amenable to cluster approaches for accurate J-coupling calculations in zeolites. DFT calculations of 2J(29Si-O-29Si) couplings of the siliceous zeolite Sigma-2, whose framework structure is known to high accuracy from single-crystal X-ray diffraction studies, yield excellent agreement between calculated and experimentally measured 2J(Si-O-Si) couplings. For the siliceous zeolite ZSM-12, calculated 2J(29Si-O-29Si) couplings based on less-certain powder X-ray diffraction analyses deviate significantly from experimental values, while a refined structure based on 29Si chemical-shift tensor analyses shows substantially improved agreement. 29Si J-coupling interactions can be used as sensitive probes of local structures of zeolitic frameworks and offer new opportunities for refining and solving complicated structures, in combination with complementary scattering, modeling, and other nuclear spin interactions. PMID- 19290356 TI - Chemistry induced by low-energy electrons in condensed multilayers of ammonia and carbon dioxide. AB - We have investigated by means of HREEL spectroscopy electron induced reactivity in a binary CO2 : NH3 ice mixture. It was shown that the interaction of low energy electrons (9-20 eV) with such mixtures induces the synthesis of neutral carbamic acid NH2COOH and that flashing the sample at 140 K induces the formation of ammonium carbamate. The products have been assigned by FTIR spectroscopy of a CO2 : NH3 mixture heated from 10 K to 240 K. A mechanism involving dissociation of NH3 molecules into NH2* and H* radicals is proposed to explain the product formation. PMID- 19290357 TI - Harnessing natures ability to control metal ion coordination geometry using de novo designed peptides. AB - Advances in protein chemistry and molecular and structural biology have empowered modern chemists to build complex biological architectures using a "first principles" approach, which is known as de novo protein design. In this Perspective we demonstrate how simple three-stranded alpha-helical constructs can be prepared by the sole consideration of the primary amino acid sequence of a peptide. With these well defined systems, we then demonstrate that metal binding cavities can be carved out of the hydrophobic cores of these aggregates in order to bind metal ions such as cadmium with well defined coordination geometries. Examples will be given of homoleptic CdS(3) complexes, CdS(3)O sites and proteins which contain equilibrium mixtures of these two species. We will provide a description of a strategy that allows us to build heterochromic peptides (small proteins that complex two metals in nearly identical environments but which result in different physical properties and allow for metal site selectivity). We conclude with a new class of designed peptides, diastereopeptides, which can exploit changes in amino acid chirality to control metal ion coordination number and lead to an alternative path towards heterochromic systems. The constructs described herein represent the initial steps of preparing protein structures that may simultaneous contain structural and catalytic metal binding centers. These studies inform the community on a developing field, which promises new opportunities for the study of bioinorganic chemistry. PMID- 19290358 TI - Self-assembly of novel 3d-4d-4f heterometal-organic framework based on double stranded helical motifs. AB - Three novel heterometal-organic frameworks {[CoCdLn(HCAM)(2)(CAM)(H(2)O)(7)].7H(2)O}(n) (Ln = Dy (); Eu (); Sm ()) have been successfully synthesized under hydrothermal condition using 4-hydroxyl-pyridine 2,6-dicarboxylic acid (H(3)CAM) as ligand, and display a fascinating 3D structure constructed by double-stranded helical motifs and exhibit luminescent properties. PMID- 19290359 TI - Synthesis and isolation of a stable, axially-chiral seven-membered N-heterocyclic carbene. AB - A chiral seven-membered N-heterocyclic carbene (NHC) has been synthesized from its phenol adduct (NHC-HOPh) by a novel base-induced alpha-elimination method, and its donor strength has been determined from the IR stretching frequencies of the NHC-Rh(CO)(2)Cl complex. PMID- 19290360 TI - A new route for substitution of the bridging acetate on the oxo-centered triruthenium acetate cluster. AB - Deacetoxylation of the oxo-centered triruthenium acetate cluster by acetyl chloride or triethylsilyl chloride led to the formation of a versatile precursor [Ru(3)O(OAc)(5)(Cl)(4-methylpyridine)(2)(CNt-Bu)], whose chloro ligand was readily substituted by other bridging ligands, i.e. alkoxides, carboxylates or sulfides. PMID- 19290361 TI - Synthesis and reactivity of the novel hydride derivative RhHCl(TIMP(3)) (HTIMP(3) = tris[1-(diphenylphosphino)-3-methyl-1H-indol-2-yl]methane). AB - The reaction of HTIMP(3) (HTIMP(3) = tris[1-(diphenylphosphino)-3-methyl-1H-indol 2-yl]methane) with [RhCl(COD)](2) and Rh(acac)(CO)(2) produces RhHCl(TIMP(3)) (1H) and Rh(TIMP(3))(CO) (2), respectively, both exhibiting tetradentate kappaC,kappa(3)P-coordination of the TIMP(3) moiety. The reaction of RhHCl(TIMP(3)) with nucleophiles (L) in the presence of AgBF(4) or AgPF(6) produces different compounds depending on the nature of L. Indeed, cationic Lewis adducts of formula [RhH(L)(TIMP(3))](+) ((2H+)-(5H+)) are obtained when L is CO, CNCH(2)Ph, pyridine or CH(2)CHCN. On the other hand, when the incoming nucleophile is CH(3)COOH the hydride-free complex [Rh(CH(3)COO)(TIMP(3))](+) ((6+)) is obtained. Finally, the reaction of RhHCl(TIMP(3)) with PhCCPh and CH(2)CHCOOMe in the presence of AgPF(6) leads to the insertion products [Rh(PhCCHPh)(TIMP(3))](+) ((+)) and [Rh(CH(2)CH(2)COOMe)(TIMP(3))](+) ((8+)), respectively. The solid state structure has been determined by single crystal X ray diffraction in selected cases (1H, (6+)). PMID- 19290362 TI - Metal-organophosphine and metal-organophosphonium frameworks with layered honeycomb-like structures. AB - Phosphanotriylbenzenecarboxylic acid (ptbcH(3); P(C(6)H(4)-p-CO(2)H)(3)) and its methyl phosphonium iodide derivative (mptbcH(3)I; {H(3)CP(C(6)H(4)-p CO(2)H)(3)}I) have been used as organic building blocks in reaction with Zn(ii) salts to obtain a series of related two-dimensional coordination polymers with honeycomb-like networks. The variable coordination number and oxidation states available to phosphorus have been exploited to produce a family of related phosphine coordination materials (PCMs) using a single ligand precursor. The phosphine carboxylate trianion, ptbc(3-), reacted with Zn(ii) to form 3,3 connected undulating hexagonal sheets based on tetrahedral P and Zn nodes, where Zn-ptbc = 1 : 1. When hydroxide was used as an additional framework ligand, Zn(4)(OH)(2) clusters were obtained. The clusters support 6,3-connected bilayers that consist of pairs of fused hexagonal sheets (Zn-ptbc = 2 : 1) with intra layer pore spaces. The Zn(4)(OH)(2) clusters are also coordinated by solvent, which was preferentially displaced when the bilayer material was synthesized in the presence of ethylene diamine. Treatment of ptbc(3-) with MeI resulted in methylation of the phosphine to give the P(v) phosphonium iodide salt derivative. The formally dianionic methylphosphonium tricarboxylate building block, mptbc(2 ), has the same trigonal-pyramidal bridging geometry as the parent phosphine. However, mptbc(2-) reacted with Zn(ii) on a 1 : 1 stoichiometric ratio to give an unusual trilayer sheet polymer that is based exclusively on 3-connected nodes. Solid-state (31)P NMR studies confirmed that the phosphine ligands were resistant to oxidation upon solvothermal reaction under aerobic conditions. PMID- 19290363 TI - Density-functional theory models of xanthine oxidoreductase activity: comparison of substrate tautomerization and protonation. AB - The hydroxylation mechanism of the molybdoprotein xanthine oxidoreductase (XOR) has been modelled using density-functional theory. High activation barriers are often obtained for models of this enzyme due to the absence of factors that stabilize the accumulation of charge on the substrate at the transition state. Xanthine provides much lower barriers than small model substrates such as formamide or imidazole due to charge delocalization to centers which appear to interact with key residues in the protein. Of the two mechanisms of stabilization discussed in the literature-tautomerization and protonation of xanthine-density functional theory calculations suggest that proton transfer from Glu1261 to N9 reduces the activation barrier by approximately 30 kcal mol(-1) and leads to an intuitive product complex. Further, similar values for the activation barriers of methyl xanthine isomers lead to the conclusion that the wide variation in rates for substituted purines is due to interactions with key residues in the active site. In addition, the transition state for oxidation of xanthine can be superimposed over the X-ray structure of inhibitor-bound XO with high correlation suggesting that the enzyme active site orients the substrate into the ideal position for reaction. The activation barriers for models of a hypothetical tungsten-substituted XO are predicted to be approximately 10 kcal mol(-1) higher in energy due to the higher reduction potential of the metal and unfavourable electrostatic interactions for the hydride transfer. PMID- 19290364 TI - Synthesis, characterisation and photochemistry of Pt(IV) pyridyl azido acetato complexes. AB - Pt(II) azido complexes [Pt(bpy)(N(3))(2)] (1), [Pt(phen)(N(3))(2)] (2) and trans [Pt(N(3))(2)(py)(2)] (3) incorporating the bidentate diimine ligands 2,2' bipyridine (bpy), 1,10-phenanthroline (phen) or the monodentate pyridine (py) respectively, have been synthesised from their chlorido precursors and characterised by X-ray crystallography; complex 3 shows significant deviation from square-planar geometry (N(3)-Pt-N(3) angle 146.7 degrees ) as a result of steric congestion at the Pt centre. The novel Pt(IV) complexes trans, cis [Pt(bpy)(OAc)(2)(N(3))(2)] (), trans, cis-[Pt(phen)(OAc)(2)(N(3))(2)] (), trans, trans, trans-[Pt(OAc)(2)(N(3))(2)(py)(2)] (), were obtained from via oxidation with H(2)O(2) in acetic acid followed by reaction of the intermediate with acetic anhydride. Complexes 4-6 exhibit interesting structural and photochemical properties that were studied by X-ray, NMR and UV-vis spectroscopy and TD-DFT (time-dependent density functional theory). These Pt(IV) complexes exhibit greater absorption at longer wavelengths (epsilon = 9756 M(-1) cm(-1) at 315 nm for 4 ; epsilon = 796 M(-1) cm(-1) at 352 nm for 5 ; epsilon = 16900 M(-1) cm(-1) at 307 nm for 6 , in aqueous solution) than previously reported Pt(IV) azide complexes, due to the presence of aromatic amines, and 4-6 undergo photoactivation with both UVA (365 nm) and visible green light (514 nm). The UV vis spectra of complexes 4-6 were calculated using TD-DFT; the nature of the transitions contributing to the UV-vis bands provide insight into the mechanism of production of the observed photoproducts. The UV-vis spectra of 1-3 were also simulated by computational methods and comparison between Pt(II) and Pt(IV) electronic and structural properties allowed further elucidation of the photochemistry of 4-6. PMID- 19290365 TI - Bulky triazenide complexes of alumino- and gallohydrides. AB - The sterically bulky triazenes DitopN(3)(H)pTol, DitopN(3)(H)Mes, DmpN(3)(H)pTol and DmpN(3)(H)Mes, where Ditop = 2,6-di-p-tolylphenyl, Dmp = 2,6-dimesitylphenyl, pTol = p-MeC(6)H(4) and Mes = 2,4,6-Me(3)C(6)H(2), have been prepared. The reactivity of these triazenide precursors with LiAlH(4) and, in the cases of DmpN(3)(H)pTol and DmpN(3)(H)Mes, LiGaH(4), with diethyl ether as the solvent, has been examined. All reactions were undertaken in a 1:1 ratio giving rise to a variety of aluminium and gallium complexes that either incorporate LiH with a metal to triazenide ratio of 1:1 or generate 'LiH-free' aluminohydrides with aluminium to triazenide ratios of 1:1 or 1:2 dependant on triazenide bulk. Increasing triazenide bulk from DitopN(3)pTol through to DmpN(3)Mes results in a transition from complexes of structure [{Li(OEt(2))(mu-H)(mu-L)AlH (mu-H)}(n)] (L = triazenide ligand; n = 2 DitopN(3)pTol, n = 1 DitopN(3)Mes, DmpN(3)pTol), to bis(triazenide) monohydride complexes [AlH(L)(2)], through to monotriazenide dihydride complexes [AlH(2)(L)]. By contrast, both DmpN(3)(H)Ar (Ar = pTol or Mes) triazenides react with LiGaH(4) to afford the monomeric, lithium hydride containing, complexes [Li(ether)(mu-H)(mu-L)GaH(2)] (L = triazenide, ether = OEt(2) or THF). The molecular structures of [AlH(DitopN(3)Mes)(2)], [AlH(DmpN(3)pTol)(2)], [AlH(2)(DmpN(3)Mes)(THF)] and [Li(THF)(mu-H)(mu DmpN(3)Mes)GaH(2)] are reported, as well as the structure of the triazene DmpN(3)(H)Mes which exists in the E-syn isomeric form in the solid-state. PMID- 19290366 TI - Electronic effects in oxo transfer reactions catalysed by salan molybdenum(VI) cis-dioxo complexes. AB - A series of molybdenum(vi) cis-dioxo complexes containing tetradentate salan ligands with different para-substitutions on the phenoxy group have been prepared. These complexes catalyse the oxygen atom transfer reaction between dimethylsulfoxide and triphenylphosphine. During oxo transfer catalysis, the complexes are resistant to formation of catalytically inactive oxo-bridged dimeric Mo(v) complexes. Electronic effects influence the rate of the oxo transfer reaction and the fastest rates are achieved when the para-phenoxy substituent is an electron withdrawing nitro substituent. Hammett correlations have shown that the rate-determining step involves nucleophillic attack of the phosphine on one of the oxo ligands. Electrochemical measurements have shown that all complexes containing tertiary amine ligands exhibit quasi-reversible behaviour and that the para-substituent has a considerable effect on the half potentials (E(1/2)). A linear correlation between the E(1/2) values and the Hammett sigma(p) parameter is observed. PMID- 19290367 TI - Adducts of the supraicosahedral stannacarborane 1,6-Me2-4,1,6-closo-SnC2B10H10; synthetic, structural and computational studies. AB - The 13-vertex stannacarborane 1,6-Me(2)-4,1,6-closo-SnC(2)B(10)H(10) reacts with the Lewis bases 2,2'-bipyridine, 1,10-phenanthroline, 4,4'-dimethybipyridine and 4,4'-diphenylbipyridine, in toluene, to precipitate bright-yellow adducts 1,6 Me(2)-4-(L(2))-4,1,6-closo-SnC(2)B(10)H(10) (, respectively) in good yield. Compounds have been characterised by NMR spectroscopy and single-crystal X-ray diffraction. Compound is somewhat less stable in solution but has also been studied crystallographically. Adduct formation retains the docosahedral structure of 1,6-Me(2)-4,1,6-closo-SnC(2)B(10)H(10) but is accompanied by polyhedral distortion arising from significant increases in Sn-C and Sn-B2 distances. This distortion is described in terms of either a slipping or hinging of the {SnL(2)} fragment. In the molecular structures of it is evident that the Sn lone pair of electrons is stereochemically active since the L(2) ligand is clearly inclined with respect to the polyhedron. Both the inclination and orientation of L(2) are rationalised by the results of DFT calculations on 4,1,6-closo-SnC(2)B(10)H(12). Calculations on models of the adducts , and reproduce the structural distortion that accompanies adduct formation, but appear to underestimate somewhat the strength of Sn-L(2) bonding. PMID- 19290368 TI - Coordination polymers of zinc with 2,2'-(2,3-bis-phenylsulfanylnaphthalene-1,4 yloxy)diacetic acid. AB - Coordination polymers of zinc with 2,2'-(2,3-bis-phenylsulfanylnaphthalene-1,4 yloxy)diacetate with different ancillary ligands such as pyridine, dimethylformamide and water are prepared and characterized. The dimensionality of zinc derived coordination polymers from (4-carboxymethoxy-2, 3-bis-phenylsulfanyl naphthalene-1-yloxy)-diacetate can be changed by ancillary ligands. In the case of pyridine as ancillary ligand it forms a one-dimensional coordination polymer, whereas in the case of dimethylformamide it is a two-dimensional polymer. PMID- 19290369 TI - A comparative study on the hydrogenation of ketones catalyzed by diphosphine diamine transition metal complexes using DFT method. AB - Three modes for hydride and proton transfer were observed in the hydrogen transfer step in ketone hydrogenation catalyzed by transition metal-centered diphosphine-diamine complexes. The results indicate that a weaker metal-hydride bond would lead to hydride transfer preceding proton transfer with a low energy barrier, and that a strong M[double bond, length as m-dash]N bond in transition metal amido complexes could increase the energy barrier for the dihydrogen activation step. PMID- 19290370 TI - Silica-bound copper(II)triazacyclononane as a phosphate esterase: effect of linker length and surface hydrophobicity. AB - A series of silica-bound Cu(ii) triazacyclononane materials was prepared to study the effect of linker length and surface hydrophobicity on the hydrolysis of phosphate esters. The general synthetic approach for these heterogeneous reagents was rhodium-catalyzed hydrosilation between an alkenyl-modified triazacyclononane and hydride-modified silica followed by metallation with a Cu(ii) salt. Elemental analysis confirmed that organic functionalization of the silica gel was successful and provided an estimate of the surface concentration of triazacyclononane. EPR spectra were consistent with square pyramidal Cu(ii), indicating that Cu(ii) ions were bound to the immobilized macrocycles. The hydrolytic efficacies of these heterogeneous reagents were tested with bis(p nitrophenyl)phosphate (BNPP) and diethyl 4-nitrophenyl phosphate (paraoxon). The agent that performed best was an octyl-linked, propanol-blocked material. This material had the most hydrophilic surface and the most accessible active site, achieving a rate maximum on par with the other materials, but in fewer cycles and without an induction period. PMID- 19290371 TI - Synthesis, structure, magnetic properties and DNA cleavage of binuclear Cu(II) Schiff-base complexes. AB - Five binuclear Schiff base copper(ii) complexes [Cu(2)(L)(OAc)].3DMF (), [Cu(2)(L)(OAc)](2).3DMF (), [Cu(2)(L)(BNPP)].3CH(3)CN (), [Cu(2)(L)(Fa)].2DMF () and [Cu(2)(L)(Pa)].DMF () (H(3)L = N,N'-bis(3,5-tert-butylsalicylidene-2-hydroxy) 1,3-propanediamine, OAc = acetic acid, BNPP = bis(4-nitrophenyl)phosphate, Fa = 2 tetrahydrofuroic acid, Pa = benzoic acid) have been synthesized and characterized by X-ray single-crystal structure analysis. Variable-temperature magnetic susceptibility studies (2-300 K) indicate the existence of ferromagnetic coupling between the copper(ii) ions in complexes and , and antiferromagnetic coupling in complexes and . The interaction of these complexes with CT-DNA has been studied by using absorption and emission spectral methods. The apparent binding constant (K(app)) values for complexes , , and are 4.67 x 10(5), 9.48 x 10(5), 4.30 x 10(5) and 3.90 x 10(5) M(-1), respectively, which show that the complexes bind to DNA by moderate intercalative binding modes. Furthermore, all these complexes can cleave plasmid DNA to nicked DNA in a sequential manner as the concentrations or reaction times are increased in the absence of reducing agent. Their cleavage activities are promoted in the presence of hydrogen peroxide. The cleavage mechanisms between the complexes and plasmid DNA are likely to involve singlet oxygen (1)O(2) and OH as reactive oxygen species. PMID- 19290372 TI - Heteroleptic ytterbium(II) complexes supported by a bulky beta-diketiminato ligand. AB - Heteroleptic beta-diketiminatoytterbium(ii) complexes [{Yb(L)(mu-I)(thf)}(2)] [: L = {N(Ar*)C(Me)}(2)CH = L(1), : L = {N(SiMe(3))C(Ph)}(2)CH = L(2); Ar* = C(6)H(3)(i)Pr(2)-2,6] were prepared by a salt elimination [from YbI(2)(thf)(2) and KL] or by a ligand redistribution [from YbI(2)(thf)(2) and YbL(2)] reaction. Complexes and were convenient precursors to some new Yb(ii) heteroleptic compounds. Thus, reaction of and with KN(SiMe(3))(2) yielded the mononuclear amido compounds [Yb(L){N(SiMe(3))(2)}(thf)] (: L = L(1) and : L = L(2)). Similarly, with K{N(H)Ar*} gave [Yb(L(1)){N(H)Ar*}(thf)] (). Complex was also obtained via a protolytic reaction of [Yb{N(SiMe(3))(2)}(2)(thf)(2)] with HL(1) at an elevated temperature. Treatment of with slightly less than a stoichiometric amount of K{CH(SiMe(3))(2)} afforded [Yb(L(1)){CH(SiMe(3))(2)}(thf)] (), while using even a small excess of the alkyl reagent resulted in the deprotonation of the beta-diketiminato ligand and the formation of the known dimer [{Yb(L(3))(thf)}(2)] [, L(3) = N[combining macron](Ar*)C(Me)C(H)C(C[combining macron]H(2))N(Ar*)]. Compounds or reacted with KCPh(3) in thf yielding [Yb(L(1)){eta(5)-C(6)H(5)CPh(2)}(thf)] () or [Yb(CPh(3))(2)(thf)(2)] (), respectively; the latter has two differently coordinated CPh(3) ligands. The molecular structures of 1, 2, 4, 5, 6, 8 and 9 were determined by X-ray diffraction. PMID- 19290373 TI - Mechanistic behaviour of alkylcobaloximes and imino-oxime complexes related to vitamin B(12). AB - The ligand substitution reactions of complexes of the type trans [(R)Co(Chel)S](+/0) with L, where chel = (DO)(DOH)pn = 2,2'-(1,3 diaminopropanebis(2-methyl-3-butanone)oxime), R = CH(3), L = imidazole, pyrazole, 1,2,4-triazole and 1-methylimidazole, and S = water and MeOH, and chel = (Hdmg)(2) = bis(dimethylglyoximate), R = CH(2)Cl, CH(2)Br, and CH(2)I, L = thiourea and pyridine, and S = water, were studied in detail as a function of temperature and pressure. The reported activation parameters (DeltaH, DeltaS and DeltaV) support the operation of a dissociative interchange (I(d)) mechanism. Complexes of the type trans-[RCo(Hdmg)(2)L] (R = CH(2)Cl, CH(2)Br, and CH(2)I; L = H(2)O and Py) were fully optimized at the B3LYP/LANL2DZp level, and the structural data support the mechanistic assignment based on the reported activation parameters. For the reaction of trans-[(CH(3))(2)Co((DO)(DOH)pn)] with acid, the activation parameters (DeltaH, DeltaS and DeltaV) were found to be 37 +/- 1 kJ mol(-1), -86 +/- 3 J mol(-1) K(-1) and -18.9 +/- 0.7 cm(3) mol(-1), respectively, and support a protonation mechanism. PMID- 19290374 TI - Role of the ancillary ligands on the stabilization of the imino-oxo tautomer of 1 methylcytosine in Pt(II) complexes. AB - The mixed nucleobases complexes cis-[L(2)Pt{1-MeTy(-H)}(1-MeCy,N(3))]NO(3) (L = PPh(3), ; PMePh(2), ), containing the N(3)-deprotonated 1-methylthymine (1-MeTy( H)) and the neutral 1-methylcytosine (1-MeCy) have been prepared and characterised. The compounds were obtained by reacting the hydroxo complexes cis [L(2)Pt(mu-OH)](2)(NO(3))(2) with 1-methylthymine (1-MeTy), followed by the addition of 1 equivalent of 1-MeCy. In solution of DMSO, DMF or chlorinated solvents, converts quantitatively into the isomer cis-[L(2)Pt{1-MeTy(-H)}(1 MeCy,N(4))]NO(3) () containing the tautomeric form of the cytosine stabilized through the coordination at the N(4) atom, as shown by single-crystal X-ray analysis. The structural determination of shows the presence in the unit cell of two crystallographic independent complexes having similar conformation, with a different orientation of the two nucleobases (head-head and head-tail) according to the presence of both isomers in solution. Complex , having the less hindered PMePh(2) ligands, in DMSO solution, contains the tautomeric forms of the cytosine in equilibrium and the migration of the metal from the N(3) to N(4) site occurs only to a minor extent. PMID- 19290375 TI - Near-infrared luminescent xerogel materials covalently bonded with ternary lanthanide [Er(III), Nd(III), Yb(III), Sm(III)] complexes. AB - A beta-diketone ligand 4,4,5,5,5-pentafluoro-1-(2-naphthyl)-1,3-butanedione (Hpfnp), which contains a pentafluoroalkyl chain, was synthesized as the main sensitizer for synthesizing new near-infrared (NIR) luminescent Ln(pfnp)(3)phen (phen = 1,10-phenanthroline) (Ln = Er, Nd, Yb, Sm) complexes. At the same time, a series of lanthanide complexes covalently bonded to xerogels by the ligand 5-(N,N bis-3-(triethoxysilyl)propyl)ureyl-1,10-phenanthroline (phen-Si) were synthesized in situvia a sol-gel process. [The obtained materials are denoted as xerogel bonded Ln complexes (Ln = Er, Nd, Yb, Sm).] The single crystal structures of the Ln(pfnp)(3)phen complexes were determined. The properties of these complexes and the corresponding xerogel materials were investigated by Fourier-transform infrared (FTIR), diffuse reflectance (DR), and field-emission scanning electron microscopy (FE-SEM). After ligand-mediated excitation of these complexes and the corresponding xerogel materials they all show the characteristic NIR luminescence of the corresponding Ln(3+) ion. This is attributed to efficient energy transfer from the ligands to the Ln(3+) ion (the so-called "antenna effect"). PMID- 19290376 TI - Use of electrochemical transient techniques to obtain thermodynamic and kinetic data on aqueous Fe(III)-1,6-dimethyl-4-hydroxy-3-pyridinecarboxylate and Fe(III) 4-hydroxy-2-methyl-3-pyridinecarboxylate complexes. AB - Voltammetric experiments were used to demonstrate the possibility to rapidly obtain stability constants, E degrees values and kinetic parameters of Fe(III) complexes with 1,6-dimethyl-4-hydroxy-3-pyridinecarboxylic acid (DQ716) at pH 2.3 and 4-hydroxy-2-methyl-3-pyridinecarboxylic acid (DQ2) at pH 3. Fe(III) diffusion coefficient (D(Fe)= 5.5.10(-6) cm(2)/s), heterogeneous electron transfer kinetic constant (k degrees = 2.7.10(-4)cm/s), symmetry coefficient (alpha= 0.57) and Fe(III)/Fe(II) standard reduction potential (E degrees = 0.53 V vs. SCE) were determined beforehand and used to obtain all the other results. Digital simulation together with potentiometric data were used to define the whole reaction system in terms of thermodynamic and kinetic parameters. In particular, E degrees and the dissociation kinetic constant, k(b), of the 1:1 (E degrees = 0.22 V vs. SCE, k(b)= 0.032 s(-1)), 1:2 (E degrees = 0.098 V vs. SCE; k(b)= 0.22 s(-1)) and 1:3 (E degrees < or =-0.29 V vs. SCE, k(b)= 157.9 s(-1)) Fe(III)/DQ716 complexes, were estimated. Stability constants of the Fe(II) complexes were computed from these values. The voltammetric data were also interpreted with two independent formalisms: (1) solution of an equation system and (2) a curve fitting method based on the Koutecky-Levich equation. Both approaches allowed us to obtain the speciation of a Fe(III)/DQ716 solution at pH 2.3. Moreover, the second approach allowed the evaluation of the kinetic contributions, the stepwise stability constant of Fe(III)L(2) (7.65 +/- 0.07), and to define the mathematical formalization of the experimental result which link some key-points of the voltammetric curve (inflection points and plateaux) to D(Fe), k degrees , alpha(j) and E degrees . This approach was also successfully applied to obtain the speciation of a Fe(III)/DQ2 solution at pH 3. PMID- 19290377 TI - Enhanced photocatalytic activity of La-doped AgNbO3 under visible light irradiation. AB - Using a solid-state-reaction method, La-doped AgNbO(3) samples of Ag(1 x)La(x)NbO(3) were synthesized and their photocatalytic activities were evaluated from gaseous 2-propanol decomposition under visible light irradiation. The photocatalytic activity and the surface area were increased by La doping, but the crystal structure and light absorption property did not change. The photocatalytic activity of Ag(1-x)La(x)NbO(3) was significantly improved over that of AgNbO(3). The Ag(0.88)La(0.12)NbO(3) sample displayed the highest photocatalytic activity: it was more than 12-fold higher than that of AgNbO(3). PMID- 19290378 TI - Biospeciation of various antidiabetic V(IV)O compounds in serum. AB - The interactions of various insulin mimetic oxovanadium(IV) compounds with serum proteins were studied in model systems and in ex vivo samples. For the modeling study, an earlier in situ method was extended and applied to the formation of ternary complexes of apotransferrin (apoTf)-V(IV)O-maltol (mal) and 1,2-dimethyl 3-hydroxy-4(1H)-pyridinone (dhp). Both systems were evaluated via simultaneous CD and EPR measurements. Determination of the formation constants of the ternary complexes allowed the calculation of more accurate stability constants for the V(IV)O-apoTf parent complexes and establishment of a better model for drug speciation in serum. It was found that dhp and the synergistic carbonate are non competitive binders. Based on the stability constants obtained for V(IV)O-apoTf complexes and estimated for V(IV)O-HSA (= human serum albumin), modeling calculations were performed on the distribution of V(IV)O among the components of blood serum. The results were confirmed by HPLC-ICP-MS (liquid chromatography inductively coupled plasma spectroscopy-mass spectrometry) measurements. The ex vivo interactions of the V(IV)O complexes formed with mal, picolinic acid (pic) and dhp with serum protein standards and also with human serum samples were evaluated. The proteins were firstly separated by (HPLC), and the V content of each fraction was determined by ICP-MS. All the studied V(IV)O compounds displayed similar chromatographic profiles, associated almost exclusively with apotransferrin as predicted by the modeling calculations. Under physiological conditions, the interactions with HSA of all of the species under study were negligible. Therefore Tf seems to be the main V(IV)O transporter in the serum under in vitro conditions, and this association is practically independent of the chemical form in which V(IV)O is administered. PMID- 19290379 TI - Solution NMR structural study of a mixed aggregate of N-lithium triphenylphosphazene and lithium bromide. AB - The toluene solution structure of an N-lithio(triphenyl)phosphazene (Ph(3)PNLi) mixed aggregate with lithium bromide (LiBr), 5, has been elucidated for the first time based on multinuclear magnetic resonance measurements ((1)H, (6)Li, (7)Li, (13)C and (31)P). The structure consist of two dimers [Li(mu-Z)](2) (Z= Br, NPR(3)) linked through LiX (X= N, Br) bridges. This arrangement is a cubane-like structure analogous to that found in the solid-state. PMID- 19290380 TI - Gestational diabetes: pathogenesis and consequences to mother and offspring. AB - Gestational diabetes mellitus (GDM) is defined as glucose intolerance with onset or first recognition during pregnancy. Data from Western countries suggest that the prevalence of GDM is increasing, being almost 10% of pregnancies and probably reflecting the global obesity epidemic. The majority of women with GDM seem to have beta-cell dysfunction that appears on a background of chronic insulin resistance already present before pregnancy. In less than 10% of GDM patients, defects of beta-cell function can be due to autoimmune destruction of pancreatic beta-cells, as in type 1 diabetes, or caused by monogenic mutations, as in several MODY subtypes. Diagnostic criteria for GDM vary worldwide and there are no clear-cut plasma glucose cut-off values for identifying women at a higher risk of developing macrosomia or other fetal complications. Because the oral glucose tolerance test (OGTT) is restricted to high risk individuals, 40% of GDM cases are left undiagnosed. Therefore, in high risk populations almost universal screening is recommended; only women considered to have very low risk do not need screening. Diet and exercise are the key elements in the treatment of GDM. If necessary, either insulin, certain oral hypoglycemic agents or combinations can be used to achieve normoglycemia. After delivery, women with GDM and their offspring have an increased risk for developing the metabolic syndrome and type 2 diabetes. Thus, pregnancy may act as a "stress test", revealing a woman's predisposition to T2D and providing opportunities for focused prevention of important chronic diseases. PMID- 19290381 TI - Stem cell therapy to treat diabetes mellitus. AB - Transplantation of pancreatic islets offers a direct treatment for type 1 diabetes and in some cases, insulin-dependent type 2 diabetes. However, its widespread use is hampered by a shortage of donor organs. Many extant studies have focused on deriving beta-cell progenitors from pancreas and pluripotent stem cells. Efforts to generate beta-cells in vitro will help elucidate the mechanisms of beta-cell formation and thus provide a versatile in vivo system to evaluate the therapeutic potential of these cells to treat diabetes. Various successful experiments using beta-cells in animal models have generated extensive interest in using human embryonic stem cells to restore normoglycemia in diabetic patients. While new techniques are continually unveiled, the success of beta-cell generation rests upon successful manipulation of culture conditions and the induction of key regulatory genes implicated in pancreas development. In this review, we compare successfully conducted protocols, highlight essential steps and identify some of the remarkable shortfalls common to these methods. In addition, we discuss recent advancements in the derivation of patient-specific pluripotent stem cells that may facilitate the use of autologous beta-cells in stem cell therapy. PMID- 19290382 TI - Combined determination of glucose and fructosamine in vitreous humor as a post mortem tool to identify antemortem hyperglycemia. AB - In clinical practice, serum glucose levels are used to diagnose diabetes mellitus. In post-mortem diagnosis, however, biochemical markers in vitreous humor are more useful because of the difficulty involved in interpreting blood glucose levels and relatively non-specific pathological features. The aim of this study was to analyze the usefulness of post-mortem determination of glucose and fructosamine combined and to compare the results with those obtained for fructosamine and combined glucose and lactate levels in two diagnostic groups (one diabetic and the other non-diabetic). We studied 377 cadavers (mean age 57.6 yr, SD 20.4, range 15 to 98 yr) with a mean post-mortem interval of 14.9 h. (SD 6.0; range 2 to 24 h). The highest levels were obtained in cases where diabetes mellitus had previously been diagnosed. In relation to diagnostic performance, the most reliable values were those in which glucose and fructosamine were determined jointly. The findings provide information concerning the usefulness of measuring glucose and fructosamine levels as a post-mortem tool for identifying antemortem glycemic control in diabetics. PMID- 19290383 TI - Posttranslational Protein Modifications in Type 1 Diabetes - Genetic Studies with PCMT1, the Repair Enzyme Protein Isoaspartate Methyltransferase (PIMT) Encoding Gene. AB - BACKGROUND: Posttranslational protein modifications have been implicated in the development of autoimmunity. Protein L-isoaspartate (D-aspartate) O methyltransferase (PIMT) repairs modified proteins and is encoded by PCMT1, located in a region linked to type 1 diabetes (T1D), namely IDDM5. AIM: To evaluate the association between genetic variations in the PCMT1 gene and T1D. METHODS: Firstly, PCMT1 was sequenced in 26 patients with T1D (linked to IDDM5) and 10 control subjects. The variations found in PCMT1 were then tested (alone and interacting with a functional polymorphism in SUMO4 and with HLA) for association with T1D in 253 families (using transmission disequilibrium test). In a third step, the association of the functional variation in PCMT1 (rs4816) with T1D was analyzed in 778 T1D patients and 749 controls (using chi-square test). In vitro promoter activity was assessed by transfecting INS-1E cells with PCMT1 promoter constructs and a reporter gene, with or without cytokine stimulation. RESULTS: Four polymorphisms in complete linkage disequilibrium were identified in PCMT1 (5' to the gene (rs11155676), exon 5 (rs4816) and exon 8 (rs7818 and rs4552)). In the whole cohort of 253 families, the allele associated with increased PIMT enzyme activity (rs4816, allele A) was less frequently transmitted to the affected than to the non-affected offspring (46% vs. 53%, p = 0.099). This finding was even more evident in the subset of families where the proband had high-risk SUMO4 (p = 0.069) or low-risk HLA (p = 0.086). Surprisingly, in the case-control study with 778 cases and 749 controls, an inverse trend was found (40.36% of patients and 36.98% of controls had the allele, p = 0.055). PCMT1 promoter activity increased with cytokine stimulation, but no differences were detected between the constructs adjacent to rs11155676. CONCLUSION: PCMT1 was virtually associated with T1D in groups defined by other risk genes (SUMO4 and HLA). A general association in a not further defined sample of T1D patients was not evident. Verification in a larger population is needed. PMID- 19290384 TI - Dose- and Glucose-Dependent Effects of Amino Acids on Insulin Secretion from Isolated Mouse Islets and Clonal INS-1E Beta-Cells. AB - BACKGROUND: The influence of glucose and fatty acids on beta-cell function is well established whereas little is known about the role of amino acids (AAs). METHODS: Islets isolated from NMRI mice were incubated overnight. After preincubation, isolated islets as well as clonal INS-1E beta-cells were incubated for 60 min in a modified Krebs Ringer buffer containing glucose and AAs. RESULTS: At 16.7 mmol/l (mM) glucose, L-arginine, L-lysine, L-alanine, L-proline, L leucine, and L-glutamine potentiated glucose-stimulated insulin secretion dose dependently, while DL-homocysteine inhibited insulin secretion. Maximal insulin stimulation was obtained at 20 mM L-proline, L-lysine, L-alanine, L-arginine (islets: 2.5 to 6.7 fold increase; INS-1E cells: 1.6 to 2.2 fold increase). L glutamine and L-leucine only increased glucose-stimulated (16.7 mM) insulin secretion (INS-1E cells: 1.5 and 1.3 fold, respectively) at an AA concentration of 20 mM. Homocysteine inhibited insulin secretion both at 5.6 mM and 16.7 mM glucose. At glucose levels ranging from 1.1 to 25 mM, the equimolar concentration of 10 mM, L-proline, L-lysine, L-arginine increased insulin secretion from mouse islets and INS-1E cells at all glucose levels applied, with a maximal effect obtained at 25 mM glucose. At a concentration of 10 mM, L-arginine and L-lysine had the highest insulinotropic potency among the AAs investigated. CONCLUSION: L arginine, L-lysine, L-alanine, L-proline, L-leucine and L-glutamine acutely stimulate insulin secretion from mouse islets and INS-1E cells in a dose- and glucose-dependent manner, whereas DL-homocysteine inhibits insulin release. PMID- 19290385 TI - Effects of Cardiorespiratory Fitness on Serum Ferritin Concentration and Incidence of Type 2 Diabetes: Evidence from the Aerobics Center Longitudinal Study (ACLS). AB - BACKGROUND: Cardiorespiratory fitness (CRF) and physical activity (PA) are inversely related to the occurrence of type 2 diabetes (T2D). Both play an important role in reducing serum ferritin (SF) concentration. Increased SF concentration is considered a contributing factor for developing T2D. METHODS: The present cohort study investigated 5,512 adult participants enrolled in the Aerobics Center Longitudinal Study (ACLS) between 1995 and 2001. The subjects completed a comprehensive medical examination and a SF evaluation, and had been followed up until either diabetes onset, death, or the cut-off date of November 2007. Three CRF levels were categorized. SF quartile levels were defined by gender and menopausal status. The incidence of T2D was calculated for 10,000 person-years, and hazard ratios (HR) were computed to predict the incidence of T2D based on SF quartiles and CRF levels. RESULTS: SF concentration was significantly higher in males than in females (148.5 +/- 104.7 ng/ml vs. 52.2 +/- 45.9 ng/ml) and was inversely associated with CRF levels. In the high CRF group, 32.7% of participants had a low SF concentration whereas only 16.8% of participants had a high SF concentration level. After adjusting for potential confounders, male participants in the highest SF quartile level had a 1.7 times (HR: 1.67, 95% CI: 1.05, 2.66; p-trend = 0.027) increased risk for developing T2D compared with those in the lowest SF quartile group. CONCLUSION: Lower SF concentration was associated with lower risk of developing T2D in those regularly participating in CRF. The findings from this study suggest that SF concentration could be used as a diabetic predictor. Based on these results clinicians and public health professionals should promote regular physical activity or fitness to reduce the incidence of T2D. PMID- 19290386 TI - Locking plates - the ultimate implant? "Nothing holds up the progress of science so much as the right idea at the wrong time." Vincent de Vigneaud, Canada (1978). PMID- 19290387 TI - Measurement of the tibial plateau angle in cats with and without cranial cruciate ligament rupture. AB - The objective of the present study was to determine the tibial plateau angle (TPA) in cats without stifle pathology and to compare it with cats suffering from an isolated cranial cruciate ligament rupture. Mediolateral radiographs of the stifle were taken and the tibial plateau angle was measured based on the method previously described by Slocum and Devine (1983) for dogs. Three observers with different levels of experience evaluated the radiographs of all of the cats in this study. The mean tibial plateau angle measured by all three observers in the cats with a rupture of the cranial cruciate ligament (CCL) was 3.1 degrees greater than in cats without stifle pathology. Neither gender, age, body weight nor degenerative joint disease had an influence on measurement results. The authors found an inter-observer variability of +/- 5.3 degrees . Hence it can be concluded that cats with cranial cruciate ligament rupture have a greater TPA, and this at least lends some credence to the possibility of higher TPA being a predisposing factor for cruciate injury in this species. PMID- 19290388 TI - The utilization of a synthetic bone void filler (JAX) in the repair of a femoral segmental defect. AB - OBJECTIVES: Currently available synthetic void fillers are indicated for bony voids or gaps that are not intrinsic to the stability of the structure. Jax TCP (tricalcium phosphate) is an osteoconductive bioceramic fabricated into 4 mm granules with a unique interlocking form, promoting structural integrity while allowing bone ingrowth. The objective of this study was to assess bone ingrowth using a large, critically sized, femoral defect. METHODS: A 5 cm segmental ostectomy was created in the mid-diaphysis of 16 adult ovine femora. A stainless steel intramedullary nail was introduced and locked with two proximal and two distal fully-threaded locking screws. Each defect was surrounded with a resorbable macroporous poly (L-lactide-co-D,L) lactide mesh acting as graft containment. Treatment groups (n = 4) were as follows: 1) Empty defect; 2) Morselized cortical bone; 3) Cortical strut; 4) Jax TCP. Serial radiographs were taken postoperatively and at two, four and six months. Femora retrieved at necropsy (six months) underwent computed tomography for volumetric analysis followed by histological assessment of the biological response. RESULTS: Little bone was apparent in the empty defect group, whereas significant bone was evident in both autograft groups and the Jax TCP group. Three-dimensional CT reconstructions and volumetric analysis were in close agreement with the radiographic findings. CLINICAL SIGNIFICANCE: Jax TCP bone graft substitute has been proven to be effective in the healing of a large, critically sized, contained segmental defect. The healing observed was superior to that of cortical struts and the new bone laid down had similar radio-opacity to autograft. PMID- 19290389 TI - Comparison of two beta-tricalcium phosphate composite grafts used for reconstruction of mandibular critical size bone defects. AB - OBJECTIVE: The aim was to compare osseointegration of blood perfused beta tricalcium phosphate cylinders (beta-TCPB) with similar composites that were additionally loaded with cancellous bone and bone marrow (beta-TCPB/BM/CB) for mandibular reconstruction. METHODS: Twelve German Black-Headed sheep with an average weight of 72.5 +/- 10 kg underwent segmental resection of the right hemi mandible. The animals that were assigned to group A (n = 6) were reconstructed using beta-TCPB while the sheep that were assigned to group B received beta TCPB/BM/CB grafts. Tissue quality was histologically assessed and bone-, scaffold , cartilage- and fibrous-tissue area were estimated using semiautomated histomorphometrical software. RESULTS: Composite grafts that were loaded with bone marrow and cancellous bone (beta-TCPB/BM/CB) exhibited significant (p<0.01) higher amounts of bone formation than beta-TCPB. The patients that were assigned to group B achieved defect union and a high grade of bone maturation. Residual ceramic remnants were rare and disconnected. Bone maturity within group A was inferior and none of the specimens showed defect union. The defect centre was still occupied by a ceramic core. CLINICAL SIGNIFICANCE: Bone and bone marrow augmented beta-tricalcium phosphate composites may qualify as a promising alternative to autograft bone for mandibular reconstruction in human and veterinary medicine. PMID- 19290390 TI - Changes to articular cartilage following remote application of radiofrequency energy and with or without Cosequin therapy. AB - OBJECTIVE: To determine the short- and long-term changes in the biomechanical properties and metabolic activity of articular cartilage following the remote application of bipolar radiofrequency (bRF) and monopolar radiofrequency (mRF) energy within the rabbit stifle joint. METHODS: The rabbits were randomly assigned to either Group-1 (normal rabbit food), or they were assigned to Group-2 (2% Cosequin in the diet). Each rabbit underwent bilateral stifle arthroscopy with either bRF or mRF applied to the infrapatellar fat pad for 45 seconds. Cartilage samples were collected at zero, four, and 14 weeks after surgery. Data were analyzed with a mixed model analysis of variance (ANOVA) for chondrocyte death, amount of GAG synthesis, and the equilibrium compressive modulus. RESULTS: A significant increase in histological damage was noted at weeks four and 14 compared to week zero. Most of the chondrocyte death noted with confocal laser microscopy (49 of 56 samples) was noted in the superficial region (outer 25%) of the articular cartilage. GAG synthesis was not significantly different between groups or devices at any time point. A significant difference was not noted in equilibrium compressive modulus throughout the study. CONCLUSIONS: Remote application of bRF and mRF energy lead to immediate chondrocyte death. Most of the damage was superficial hence the metabolic activity and biomechanical properties of the extracellular matrix were maintained throughout this study. Treatment with Cosequin did not prevent superficial chondrocyte death caused by the application of radiofrequency (RF) energy with in the joint. PMID- 19290391 TI - Effects of radial extracorporeal shock wave therapy on radiographic and scintigraphic outcomes in horses with palmar heel pain. AB - OBJECTIVE: To investigate the effects of radial extracorporeal shock wave therapy (RESWT) on radiographic and scintigraphic variables in horses with clinical pain referable to the palmar heel. METHODS: Eight client-owner horses with palmar heel pain were treated with RESWT for a total of three treatments. Nuclear scintigraphy and radiography were repeated at the beginning and completion of the study. Scintigram region of interest (ROI) density ratios were calculated and compared between treated limbs, untreated limbs, and a population of comparison limbs from eight horses free of lameness. Radiographs were scored for whole navicular bone appearance as well as distal border synovial fossae number and severity. RESULTS: There was not any pre- versus post-treatment difference in scintigraphic navicular pool phase or delayed phase ROI density ratios in treated limbs, or between treated and untreated limbs. Delayed phase ROI density was increased in the central navicular region in treated limbs compared to comparison limbs from non-lame horses at both time points. Radiographic scores remained unchanged. CLINICAL SIGNIFICANCE: RESWT as applied in the present study has no effect on acute palmar heel region scintigraphic or radiographic parameters. Any acute clinical benefit may be due to analgesic effects rather than stimulation of local tissue metabolism. PMID- 19290392 TI - Morphological and biomechanical studies on the common calcaneal tendon in dogs. AB - Spontaneous rupture at the distal part of the gastrocnemius tendon (GT) is the second most common non-traumatic tendon injury in dogs, whereas the other strands of the common calcaneal tendon do not seem to have a predisposition to rupture. In order to discover why we investigated the common calcaneal tendons of 63 dogs microscopically and biomechanically. Both the gastrocnemius and superficial digital flexor tendon (SFT) had multiple low vascularized fibrocartilaginous areas within their distal course as opposed to regular parallel fibered areas in the proximal tendon areas. Biomechanical testing revealed that the distal sections in both tendons show a 50% and 70% lower tensile strength (F(max)/kg BW) than the proximal sections (p<0.01), respectively. On the contrary, tensile load (F(max)/mm(2)) only differed minimally between proximal and distal sections in both tendons (8% and 9%, respectively), whereas the tensile load of the distal gastrocnemius tendon is 35% lower than of the distal superficial flexor tendon (p<0.01). To the authors' knowledge, this is the first study to experimentally show that there are different biomechanical properties within the same tendon. The maximum load to failure is lower in the GT compared to the SFT within the same dog which explains its higher incidence of rupture in the field. The avascular fibrocartilaginous structure in the distal gastrocnemius tendon seems to play a further role in the pathogenesis of spontaneous rupture. PMID- 19290393 TI - The effects of screw placement on plate strain in 3.5 mm dynamic compression plates and limited-contact dynamic compression plates. AB - The objective of this study was to evaluate the effect of screw omission on plate strain during axial load to failure and cycling using a Delrin rod gap model. In addition, the differences between the 3.5 mm limited-contact dynamic compression plate (LC-DCP) and the 3.5 mm dynamic compression plate (DCP) were evaluated. Six, 12-hole LC-DCP and DCP plates were applied to Delrin rods with a 1 cm gap located within the central portion of the plate. Six screw configurations were tested with the following variations: three proximal and three distal (six open), four proximal and three distal (five open), four proximal and four distal (four open), five proximal and four distal (three open), five proximal and five distal (two open), six proximal and five distal (one open). Three strain gauges were mounted on each plate within the gap (gauge three) and extended proximally. Additionally, three constructs (six, three, and one open hole) were tested to failure in cyclic loading. The strain measured within the gap (gauge three) was significantly greater than the strain at other gauges for each screw configuration. Strain within the gap did not significantly change with any screw configuration, but did significantly increase at other locations as screws were omitted. Overall, the DCP withstood significantly more cycles than the LC-DCP. Differences were noted within the DCP group with the 6/5 screw configuration lasting for significantly more cycles than the 5/4 and 3/3 constructs. Although overall strain at the gap did not significantly increase with screw omission, the clinical significance remains to be determined. PMID- 19290394 TI - The effect of bending and twisting on the stiffness and strength of the 3.5 SOP implant. AB - Locking plates are becoming increasingly popular in veterinary orthopaedics. The SOP is a novel locking plate system, which can be contoured with 6 degrees of freedom and which utilises standard bone screws. The purpose of this work was to investigate the mechanical consequences of contouring the 3.5 SOP plate to support the formulation of clinical guidelines. The implants were loaded in four point bending using an industry standard protocol. The uncontoured SOP was found to be significantly stiffer and stronger than the uncontoured 3.5 DCP. Bending, and to a lesser extent, twisting, diminished the SOP's stiffness and strength but the contoured SOP remained at least as stiff and strong as the untouched DCP. PMID- 19290395 TI - Ground reaction force analysis of unilateral coxofemoral denervation for the treatment of canine hip dysplasia. AB - Coxofemoral denervation has success rates of 90-96% reported retrospectively for palliative treatment of hip dysplasia. The aim of this study was to objectively evaluate ground reaction forces (GRF) in dysplastic dogs after unilateral denervation. Unilateral coxofemoral denervation was performed by means of a previously reported technique on 10 dogs with asymmetric gait. GRF were measured at zero, one and three months. Statistical analysis was performed using repeated measures analysis of variance and paired t-tests, with p< or =0.05. There was a lack of significant difference in mean peak vertical force (PVF) or vertical impulse (VI) in the operated limb (TX) over time. For the unoperated limb (UnTX), mean PVF and VI significantly decreased over time. The dogs were significantly more lame in the TX limb initially and at one month, however, there was no significant difference between limbs by three months. In the UnTX limb, 40% of dogs decreased PVF by >5%, by three months. Over time, there was no significant difference in mean average rise or mean average fall for TX or UnTX limbs. Between limbs, mean rise in the TX limb was significantly less at zero months, but not at one or three months. Decreased compensatory load shifting to the UnTX limb due to procedural efficacy could explain decreases in the UnTX limb. Worsening disease could also explain decreases in the UnTX limb, and may indicate a protective effect denoted by a lack of change in the TX limb. Longer follow-up would be required in improved dogs in order to document continued efficacy. PMID- 19290396 TI - An evaluation of prosthetic femoral head impact on acetabular articular cartilage in a hemiarthroplasty model. AB - OBJECTIVES: The quantitation of the impact of hemiarthroplasty currently involves clinical outcome measures and radiographic evaluation. This study used in vivo and in vitro means in order to evaluate articular cartilage changes following hemiarthroplasty in the sheep. METHODS: Radiostereometric analysis (RSA) was utilized in order to quantitate migration (penetration) in vivo of the femoral head into the articular cartilage of the acetabulum. Dynamic stiffness of retrieved specimens was quantitated in vitro using a handheld device. Chondrocyte viability was assessed using live/dead cell staining and quantitated using image analysis techniques. Saffranin-O staining provided histological assessment of the cellular response to hemiarthroplasty. RESULTS: RSA showed rapid penetration of the femoral head into the acetabular cartilage over six months. Significantly higher levels of dynamic stiffness were observed in the acetabula following hemiarthroplasty. Confocal imagery highlighted changes in chondrocyte distribution and morphology. A loss of metachromasia, fibrovascular tissue invasion and apoptosis were observed histologically. CONCLUSIONS: RSA and measurement of the dynamic mechanical characteristics of cartilage provide a means of evaluating the development of degenerative articular cartilage changes in response to hemiarthroplasty. PMID- 19290397 TI - Rehabilitation after extra-articular stabilisation of cranial cruciate ligament rupture in dogs. AB - Patients at the Blue Star Foundation animal hospital in Gothenburg, Sweden, underwent a procedure with an extracapsular suture technique for cranial cruciate ligament rupture. Two groups were compared; one of the groups received rehabilitation by the owner, and the other received professional rehabilitation by a trained physiotherapists. The dogs where reexamined at four, 12 and 24 weeks postoperatively. The results were 'good' in both groups but a difference was not seen between the groups. This study shows that in the dogs with a body weight of between 25 and 50 kg with cranial cruciate ligament rupture, the extracapsular suture technic gives good results, provided that the dogs are actively walked on the leash postoperativly. PMID- 19290398 TI - Initial experience with a newly developed cementless hip endoprosthesis. AB - The HELICA-Endoprosthesis is a newly developed cementless hip prosthesis for dogs. It was implanted in 39 dogs that had severe hip osteoarthritis and a history of hip pain, as well as in one dog that had chronic hip luxation. One dog had a bilateral arthroplasty. The body weight of the patients ranged between 22 and 54 kg and their ages between nine months and 10 years. Both the femoral stem and acetabular component of the prosthesis were screwed into position following bony preparation. Additional fixation was not necessary as the components remain fixed in position until osteointegration is complete. There are currently five sizes of prosthesis available, and the various components such as the stem, cup and head are readily interchangeable. Although it appeared that good osseous anchorage of the prostheses in the bone on the surgery table had been obtained, three patients experienced both stem and cup loosening (one week, three weeks and six months after surgery). In one animal, stem loosening was observed six weeks after surgery, and another dog experienced a cup loosening two weeks postoperatively. Most of the complications were due to technical errors that occurred during the learning phase. Surgical revisions were successful in three out of five animals. In two animals we had to perform a femoral head and neck excision. Two other animals experienced radiographic bone resorption underneath the segmented collar of the femoral prosthesis but did not show any significant clinical signs of lameness. Another dog that showed signs of ischial neuropraxy after surgery, recovered completely within six weeks after surgery. All of the dogs were capable of weight bearing on the operated leg one day after surgery. The main advantage of the HELICA-Endoprosthesis is the relatively easy surgical technique and short surgery time. The initial clinical results in these 40 cases have been very encouraging. The aim of this study was to assess the early clinical results in these 39 dogs. Final evaluation can only be based on the results of gait analysis, long-term follow-up, and post mortem histological analysis. PMID- 19290399 TI - Ilio-femoral external fixator application for temporary stabilization of recurring caudal ventral hip joint luxation after total hip replacement. AB - An ilio-femoral external fixator was applied to a 13-year-old male, castrated mixed breed dog with a caudal ventral luxation that persistently luxated following both closed reduction and open reduction with a total hip replacement. The external fixator was placed in order to temporarily maintain reduction during soft tissue healing. Upon removal of the external fixator the dog underwent physical therapy. Reduction was maintained and the dog returned to normal function. PMID- 19290400 TI - Adhesions of small intestine and urinary bladder to fracture callus within the pelvic canal in two dogs. AB - In this case report, we report on two cases of pelvic fractures with uncommon complications that resulted from adhesions of the small intestine to the fracture callus in dogs. The most likely cause of the clinical signs in the first case was adhesions between a segment of the jejunum and a coccygeal callus which resulted in luminal narrowing. This was further complicated by small obstructing trichobezoars. In case 2, which was admitted with incontinence, adhesions were found between fractured pubic bone callus and both the urinary bladder and a segment of the jejunum. PMID- 19290401 TI - Arthrodesis of the tarsometatarsal joints in a cat with a two hole advanced locking plate system. AB - This report describes the successful use of the novel fixed-angle locking plating system ALPS (advanced locking plate system) in the case of dorsal tarsometatarsal instability in a cat. Partial arthrodesis of the tarsometatarsal joint was performed with a two hole ALPS 5 and two 2.4 mm selftapping locking screws through a small dorsal approach. Five days after the surgery the cat was walking without lameness. Radiographs made after six weeks showed uneventful healing. ALPS offers an alternative to existing plating systems when selecting an implant for the stabilisation of dorsal tarsometatarsal instabilities in cats. PMID- 19290402 TI - Chronic intervertebral disk herniation associated with fused vertebrae treated by vertebral lateral corpectomy in a cat. AB - A 10-year-old Domestic Shorthair cat was admitted for chronic ambulatory paraparesis and a spinal malformation. The clinical examination revealed paraparesis accentuated on the left side. Thoracolumbar radiographs revealed a spinal malformation with a narrowed intervertebral space between L1 and L2, and a dorsal fusion at the level of L2-L3 with a common dorsal process. Magnetic resonance imaging (MRI) revealed an intervertebral disk herniation with a ventral compression of the spinal cord at the level of L1/2. A standard vertebral lateral corpectomy with a foraminotomy was performed with a good outcome. PMID- 19290404 TI - [History of scientific truth: from Determinism, through Indeterminism to Probabilism]. AB - In this article it is reported a historical-philosophical "excursus" on the schools of thought followed by science in order to provide the highest degree of truth to the knowledge of natural reality. PMID- 19290405 TI - [Early diagnosis of dysmorphophobia and others dysmorphic disorders: a possible operative model]. AB - AIMS: Body Dismorphic Disorder interest the 1-2% of the general population. It is characterized by an unfavorable prognosis and an elevated comorbidity with others psychiatric disorders. Approximately 6-15% of people with body image disorder refer for aesthetic surgery without any benefit indeed the outcome is, often, to request others surgery procedure. The aim of study has been to investigate the presence of dismorphophobia, others dismorphic disorders and psychiatric symptoms in a sample of patients candidates to aesthetic surgery procedure. MATERIALS AND METHODS: It has been recruited 109 patients of mean age 27.21 years, coming to the department of Plastic Surgery of L'Aquila (Italy). To all the patients have been delivered a questionnaire for socio-demographic data and two clinical standardized instruments: Self-report Symptom Inventory-Revised (SCL-90) and the Body Uneasiness Test (BUT). The psychometric and clinical evaluations have been performed by SMILE center (Service for Monitoring and early Intervention against psychoLogical and mEntal suffering in young people), the mission of which is to reduce the burden of mental suffering in young people by of an earlier recognition of signs and symptoms of psychiatric disorders. RESULTS: 42.67% of sample showed positive results to BUT, while 57.33% reported negative results. Moreover, BUT-positive subjects achieved significantly higher results in all SCL 90 items. CONCLUSIONS: A liaison between psychiatrists and plastic surgery equipe is essential for an early diagnosis of Dismorphophobia and others Dismorphic Disorders, to begin an eventual treatment and to supply indications for the surgical candidability. Our study confirm the high prevalence of body image disorders and the presence of other psychiatric symptoms and diseases in patients who demand aesthetic procedure. The detection of early signs of any psychiatric and psychological apparent discomfort represent a priority because this correlate, in most cases, with late diagnosis and intervention because awareness of these types of disease is usually very low and therefore the possibility that subjects with this type of problems are requested to psychiatric services is rare. PMID- 19290406 TI - Treatment of hyperhomocysteinaemia in haemodialysis patients at high cardiovascular risk. AB - OBJECTIVES: Cardiovascular disease (CVD) is the main cause of death among haemodialysis (HD) patients. Emerging cardiovascular risk factors such as oxidative stress and chronic inflammation are involved in these patients together with traditional risk factors. Here we investigate the effects of a short-term folate treatment on some markers of chronic inflammation in two groups of HD patients with and without vascular occlusive disease (VOD). PATIENTS AND METHODS: Homocysteine (HCy), C-reactive protein (CRP), Folate, fibrinogen and alpha1 acid glycoprotein (alpha1AGP) were dosed before and after a 3-month course of high dose folate (25 mg intravenous calcium laevofolinate pentahydride once weekly) and again after a one-month washout in 15 HD patients with established VOD (group A) and in 15 comparable HD patients with no diagnosis of VOD (group B). RESULTS: Baseline HCy and CRP were significantly elevated in patients of both groups A and B compared to normal values. Folate treatment significantly reduced HCy in patients of both groups A and B and alpha1AGP only in patients of group A, while the other markers were not modified. After the one-month washout a significant raise of CRP could be observed in patients of group A; again, the other markers were not modified. CONCLUSIONS: Our results suggest that significant reduction of serum HCy can be achieved in both patients with or without VOD after administration of high-dose folic acid. Hence, folic acid supply is useful in the treatment of hyperhomocysteinemia in HD patients, although it is not sufficient to modify their chronic inflammatory status. PMID- 19290407 TI - [Clinical study on 40 cases of inflammatory pathologies of upper respiratory and digestive tract treated by inhalatory crenotherapy]. AB - OBJECTIVES: Authors present beneficial effects of crenotherapy on 40 patients suffering from inflammatory mucosal conditions of upper respiratory-digestive tract (URDT). MATERIALS AND METHODS: Our study was realized at the hydrothermal premises in Telese Terme (BN, Italy). Subjects selected for this study are 40 patients (22 males and 18 females) aged 20-68 years (mean 38.6 years) who suffered from catarrhal and inflammatory mucosal diseases of URDT. The patients, who are informed about the modalities of the study, undergo E.N.T. examination and nasal mucosal brushing for cytologic analysis before and after crenotherapy. RESULTS: At the end of the therapeutic course we observed a relevant improvement of clinical indicators and of mucociliary transport time in URDT. Moreover, a satisfactory control of local inflammation is highlighted by a rhinocytogram, performed after crenotherapy and showing an increase in plasma cells, a decrease in granulocytes and a normalization of mucous secretion (nasal mucosal histology). CONCLUSIONS: Our study shows that sulphurous waters are particularly effective in anti-catarrhal and anti-inflammatory therapy of URDT. PMID- 19290408 TI - [Medical-surgical integrated approach in the treatment of non-paraneoplasic hyperparathyroidism: our experience]. AB - OBJECTIVE: Hyperparathyroidism is a generalized alteration of calcium, phosphorus and bone metabolism due to an increased secretion of parathyroid hormone (PTH). In addition to the paraneoplastic ectopic type, we can distinguish three eutopic types of hyperparathyroidism, i.e., the primary form, mostly due to a benign or malignant tumor of parathyroid gland, the secondary form, typical of kidney disease and tertiary form, due to the progression of secondary forms. There is not agreement, in medical literature, on the treatment of these patients. To establish the correct therapeutic approach in patients with hyperparathyroidism, we have followed a group of symptomatic subjects suffering from primary, secondary and tertiary hyperparathyroidism, taking into account the therapeutic needs. MATERIALS AND METHODS: We followed for 12 months 155 patients suffering from primary, secondary and tertiary hyperparathyroidism; 82 were in end stage kidney disease, 93 were hypertensive. Subjects with primary forms has been treated, before parathyroidectomy, with idration (physiological solution of NaCl), bisphosphonates i.v. (pamidronate 60-90 mg in 4-6h) and, if serum calcium was higher than 12 mg/dl, loop diuretics (furosemide 40 mg/day). Subjects with secondary forms has been treated with hypophosphoric diet, phosphate bindings (calcium carbonate 1 g/day) and oral calcitriol (1 microg/d) before subtotal parathyroidectomy. After surgery it was administered support therapy with calcium gluconate (40 ml/day) and vitamin D (2.5mg/d) until serum calcium normalization. RESULTS: There were 55 cases of post surgery hypertensive attack treated with clonidine (300 microg/d); 8 months later there was not relapses but in all patients there was reduction of serum calcium concentration that required a substitutive treatment (calcium 1 g/day and calcitriol 1 microg/day). There was 1 case of heavy hypocalcemic state treated with calcium gluconate i.v. (40 ml/day). CONCLUSIONS: A correct approach to a non-paraneoplastic hyper-parathyroid patient need of an integration of both current medical and surgical options. In primary forms the first option is the surgical approach supported by medical treatment. In secondary forms medical approach is preferable to control renal and vascular complications, while surgical therapy is to prefer in non-responders to medical therapy forms. PMID- 19290409 TI - An unusual case of additional branches of median nerve innervating the corachobrachialis muscle and its clinical implications. AB - The flexor compartment muscles of the arm comprising of biceps brachii, brachialis and the coracobrachialis are innervated by the musculocutaneous nerve arising from the lateral cord of the brachial plexus. In the present study, we report a case of anomalous innervation of the corachobrachialis muscle on the left side of a 45-year-old male cadaver. The musculocutaneous nerve originated from the lateral cord, as usual and pierced the corachobrachialis muscle. The median nerve was formed by a contribution from both lateral and medial roots, both of which took origin from the lateral and medial cords, respectively. In addition to the usual musculocutaneous nerve which pierced the corachobrachialis muscle and innervated it, two more anomalous branches from the median nerve were observed to innervate the corachobrachialis. The anatomical knowledge of the variations of the innervations of the corachobrachialis muscle may be important not only for surgeons performing coracoid transfer but also for clinicians diagnosing nerve lesions. PMID- 19290410 TI - [Is body surface area (BSA) suitable for indexing renal and dialysis clearances and other physiological parameters? A theoretical approach and a critical analysis (Part 1)]. AB - Many parameters of biological functions, interesting different branches of medicine, have to be indexed by the same value for each other comparison between single individuals or between-within groups. Body Surface Area (BSA) and 1.73 m2 reference value are probably the most used tools for indexation, but a growing number of scientific papers emphasize the risk of under or over estimates of data by BSA indexation in overweight--obese or on the contrary underweight individuals. This study has been planned to evaluate this problem: a) by a theoretical approach, indexing to 1.73 m2 the creatinine clearance (CtCl) in an ideal population and b) analysing and debating the doubts and problems about the indexation by BSA in many fields, the possibilities of indexation by other methods, or to use BSA with different approach, to reduce the risk of bias, evaluating the possibility of this target in a real population of 45 men and 84 women undergoing Peritoneal Dialysis (PD). The effects of indexing 1.73 m2, according different weights on the CtCl of the ideal population and on the values of BSA estimation according different formulae. Results, methods, and the problems due to abnormal fluctuation of weight compared to normal range of standard value according height are analyzed and discussed. PMID- 19290411 TI - [Fiberoptic bronchoscopy as diagnostic tool in primitive lung cancer]. AB - OBJECTIVE: The aim of this study was to assess the diagnostic role of fiberoptic bronchoscopy in primitive lung cancers in relation to the sensibility, the specificity and the cost of the examination. MATERIALS AND METHODS: We analysed the diagnostic flow-charts of 473 patients with suspected primitive lung cancer consecutively examined during a period of 4 years (2003-2006). The results were analysed comparing patients observed in the period 2003-2004 with those observed during 2005-2006. The number and type of samples collected and the protocols utilized were considered. RESULTS: In overall patients the reliability of the fiberoptic bronchoscopy was 61.7%, with a significant increase from 47.5% in 2003 2004 to 74.4% in 2005-2006. An important role in improving the diagnostic relevance of the exam was assumed by the more frequent adoption of biopsies and trans-bronchial needle aspiration on parenchyma and mediastinal lymph nodes. CONCLUSIONS: The fiberoptic-bronchoscopy associated to advanced tissue sampling techniques represents the gold standard for the diagnosis of lung cancer, due to high sensitivity and specificity and moderate cost. PMID- 19290412 TI - [Antithyroid drugs therapy]. AB - Methimazole and Propylthiouracil are the cornerstones in the management of Graves' disease. Their primary effect is to inhibit thyroid hormone synthesis at different steps, i.e. in thyroid gland and in peripheral tissues. Antithyroid drugs can be used as the primary treatment for hyperthyroidism (long term therapy: 1-2 years) or as preparative therapy before radioiodine treatment or surgery (short term therapy: weeks or months). Generally, the starting dose of methimazole is 10-30 mg, as single daily dose, while that of PTU is 100-300 mg every 6 hours. Methimazole is the drug of choice, because major side effects are less common, it can be used as single dose, it's less expensive and more available. As far as the treatment of hyperthyroidism in pregnancy, MMI and PTU have same therapeutic efficacy and are both safe for the fetus, having similar placental transfer kinetics. The use of methimazole can be associated with aplasia cutis and choanal/esophageal atresia, while there are no data supporting the association between congenital anomalies and PTU. For this reason the latter is the drug of choice in the treatment of hyperthryroidism in pregnancy. Both thionamides are present in breast milk, but there are no controindications for their use during lactation. PMID- 19290413 TI - [Emergency Radiology layout]. AB - The purpose of Radiologic Unit in Emergency is to reach diagnostic and therapeutic effectiveness in the best way and in less time possible. The Portable Ultrasound Device is an instrument necessary in Emergency Room and in ambulance/helicopter to evaluate the evidence of endoperitoneal bleeding. The CT is the radiologic methodic more fast that permits a complete evaluation of all body segments in traumatized patient in the famous "golden hour" after the trauma, therefore it would be placed in Emergency Area. The multislice CT brought to a reduction of morbidity and mortality, thanks to a quick acquisition, to a thin collimation, to a more spatial resolution and to an optimal vessel opacization, determining a saving of hospital global costs, therefore a reduction of percentage of not necessaries operations and permitting a more rapid diagnosis, obtaining a considerable reduction of waiting in Trauma Emergency Room with more rapid and aimed therapies and a consequent costs reduction. To satisfy a so wide question of radiologic exams necessaries devices are informatic systems completely connected between Radiology department and other departments. Main advantages of MR in Emergency are the use of non ionising radiations, the possibility to effect diffusion and perfusion studies and to evaluate spinal cord damage. Reduction of time of patient preparation and times of acquisition and elaboration of imagines by modern and performant devices is basic to make more rapid therapeutic decisions. PMID- 19290414 TI - [Imaging in diagnosis of acute pancreatitis and its complications]. AB - Pancreatitis is a flogistic disease, caused by activation and digestion of pancreas by its enzymes. Diagnosis is based on integrated evaluation of clinical and laboratoristic data and morphological imaging. To evaluate the severity of pancreatitis there is a clinical classification in interstitial--mild pancreatitis and severe--necrotic one. The evaluation of severity is basic, because it is strictly correlated to the prognosis of the patient. CT has revealed the best method for diagnosis, staging and for evaluate the complications and follow-up and in some cases it is useful for therapeutic change.The abdomen X-Ray in orthostatism is performed in every situation suspected for acute abdominal disease, also if aspecific; the ultrasound can be used as first instance method in patient with clinical suspect of acute pancreatitis; the MR has actually a secondary role for the diagnosis, with only except for dubious cases to exclude primitive tumor of pancreas and pancreatic shock, but it represents, instead, first instance method in patients with adverse reaction to contrast medium. The CPRE has, like angiography, a selective indication. PMID- 19290415 TI - Modern therapeutic approach to ANCA-associated vasculitis. AB - The systemic vasculitis are characterised by an inflammatory process of the blood vessel walls. These may affect any of the vessels and organs, resulting in a diverse range of signs and symptoms. These polymorphic clinical manifestations, combined with the non-specific aetiology of the histological lesions, render the diagnosis for any single form of vasculitis more complicated. The problem lies in the various forms of vasculitis with analogous clinical presentations, requiring very different prognosis and treatment. This review has been realised in order to examine the modern approach to achieve and to maintain the clinical remission and to treat the relapse. PMID- 19290416 TI - Alternative tools for field analysis on malarial infection: a reappraisal. AB - Malaria is an important tropical infection affecting millions of world population. The gold standard for diagnosis of malaria is finding for intraerythrocytic malarial parasite from peripheral blood smear examination. Field diagnosis is important in disease detection and control. There is a need to find new alternative diagnostic tool for field diagnosis for malarial infection. The author hereby briefly reviews on the alternative diagnostic test for diagnosis for malaria in field study. PMID- 19290417 TI - [The correlation between interleukin-1 receptor antagonist genotype and coronary heart disease in patients with chronic periodontitis]. AB - PURPOSE: To investigate the correlation between interleukin-1 receptor antagonist (IL-1ra) genotype and chronic periodontitis in patients with coronary heart disease. METHODS: Genomic DNA was obtained from buccal swabs in 86 patients with coronary heart disease,of which 45 patients with chronic periodontitis, the other without chronic periodontitis. Genotypes of IL-1RN intron 2 VNTR were analyzed by SSP-PCR method. Then the differences in distribution of each genotype were analyzed with SPSS 13.0 software package. RESULTS: A significant over representation of IL-1 RN intron 2 VNTR allele 2 was found in patients with coronary heart disease and chronic periodontitis group (chi(2)=4.960,P=0.026). CONCLUSION: IL-1RN intron 2 VNTR allele 2 may be a risk indicator for the susceptibility of chronic periodontitis in patients with coronary heart disease. PMID- 19290418 TI - [Sagital position of glenoid fossa in Angle Class II malocclusion with mandibular retrusion]. AB - PURPOSE: To evaluate and compare the morphology and position of the glenoid fossa in patients with Class II malocclusion associated with mandibular retrusion and Class I malocclusion in the permanent dentition. METHODS: A sample of 26 subjects(12 males, 14 females, average age 13.4 years) with skeletal and dental Class II malocclusion associated with mandibular retrusion, was compared with a matched group of 31 subjects(12 males, 19 females, aged 12.9 years) with skeletal and dental Class I relationships. The lateral cephalograms were traced and the cephalometric analysis was taken by Nemotec Dental Studio 2006 software before orthodontic treatment. The comparisons between the two groups on the cephalometric measures for the assessment of glenoid fossa were performed by means of an independent samples t test using SPSS 15.0 software package. The correlation and regression analysis was also performed within the measures in the Class II group. RESULTS: The results showed there was significant difference of Wits, GF/Ptm-FH and GF-FMN between the two groups (P<0.01). There was a significant correlation between the GF-FMN vs. AOBO, GF-FMN vs. GF/S-FH, GF/S-FH vs.GF/Ptm-FH, S/Ptm-FH vs. GF/Ptm-FH in the Class II group. The regression equations of the GF-FMN vs. AOBO and GF-FMN vs. GF/S-FH were also obtained (P<0.05). CONCLUSIONS: The Class II malocclusion associated with mandibular retrusion patients have a posteriorly displaced glenoid fossa compared with Class I patients. Cephalometric measurement is a useful tool to evaluate the glenoid fossa morphology and position. PMID- 19290419 TI - [Quantitative evaluation of residual endodontic microorganisms after mechanical root canal preparation different chemical preparations by real-time PCR]. AB - PURPOSE: To establish a quick, sensitive method for quantifying root canal flora and investigate the effects of different root canal preparations on the pathogenic bacteria at RNA level. METHODS: A total of 24 single-rooted teeth with chronic apical periodontitis were selected and prepared using 3% H2O2 combined with 1% NaClO, EDTA combined with 3% H(2)O(2),1% NaClO, respectively,the samples were taken before and after root canal preparation. After isolation of total RNA from the root canal samples, cDNA was synthesized by reverse transcription, and detected by real-time PCR. The data were analyzed with SAS 6.12 software package. RESULTS: The number of bacteria in the root canal reduced dramatically after mechanical preparation and irrigated using 3% H(2)O(2) and 1% NaClO(P<0.01). Further combined with EDTA, its effect was better than that of simply irrigated using 3% H(2)O(2) and 1% NaClO(P<0.05). CONCLUSIONS: Real-time PCR can be employed in the identification of bacteria flora in the root canal, both methods of root canal preparation can effectively reduce the number of bacteria flora. PMID- 19290420 TI - [Combined surgical and orthodontic treatment of severe maxillary anterior teeth and dentoalveolar bone protrusion]. AB - PURPOSE: The purpose of this study is to investigate the outcome and stability of combined surgical and orthodontic treatment of severe maxillary anterior teeth and dentoalveolar bone protrusion. METHODS: Twenty-eight adults with severe bimaxillary protrusion were included.Presurgical orthodontic treatment coordinated axis of upper anteriors and dentoalveolar bone.Maxilla was impacted and set back by Le Fort I osteotomy and maxillary anterior segmental osteotomy, while upper anteriors and dentoalveolar bone were uprighted and retracted. Post surgical orthodontic treatment detailed the occlusion.Each patient underwent X ray examination at five time points including pre-treatment, pre-surgery, one month after surgery, three months after surgery and post-treatment. Hard tissue and soft tissue changes were investigated by cephalometric analysis. The data were analyzed by paired t tests with SPSS12.0 software package. RESULTS: It was observed that upper anteriors were uprighted and retracted (11.77+/-9.9) degrees (P<0.001) after treatment while dentoalveolar bone was uprighted (6.24+/-3.9) degrees (P<0.001). UI-LI angle increased (19.4+/-8.6) degrees (P<0.001), UI-Apog distance decreased (6.62+/-2.2)mm (P<0.001). The thickness of alveolus on anterior teeth labial side showed no change (P>0.05).The treatment outcome demonstrated a significant improvement in soft tissue profile. Nasolabial angle increased (11.06+/-11.8) degrees (P<0.001), upper and lower lips moved backward, and pog' moved forward. Most measurements showed no significant difference from surgery time to post treatment. CONCLUSIONS: Combined surgical and orthodontic treatment is a very effective procedure in treatment of severe maxillary anterior teeth and dentoalveolar bone protrusion and gummy smile. It remains the thickness of alveolus on anterior teeth labial side. It shows big improvement in soft tissue profile and great stability. PMID- 19290421 TI - [Diagnostic value of magnetic resonance sialography for submandibular sialolithiasis]. AB - PURPOSE: To evaluate the role of MR Sialography(MRS) in diagnosis of submandibular sialolithiasis. METHODS: Twenty-one patients with submandibular sialolithiasis were scanned by MRS, the results of MRS were compared with those after proper treatment. RESULTS: The results of MRS were consistent with those after proper treatment. CONCLUSION: MRS has high diagnostic accuracy in diagnosis of sialolithiasis including submandibular sialadenitis. PMID- 19290422 TI - [The relationship between sagittal upper airway size and surrounding skeletal structure with Delaire cephalometric analysis]. AB - PURPOSE: To investigate the relationship between sagittal upper airway size and craniofacial characteristics in men with malocclusion in early permanent dentition,and to find out the factor that affect the pharyngeal airway size remarkably. METHODS: Seventy-four cephalometric films of non-snoring males with malocclusion aged from 11 to 16 years(mean 13 years) before treatment were collected and measured with Delaire cephalometric analysis,the results were assessed by SPSS13.0 software package,multiple regression analysis was used to examine the relationship between upper airway size and craniofacial characteristics variation. RESULTS: The size of nasopharyngeal space (PNS-ad) correlated negatively with the upper face height,the inclination of ramus plane,percentage of cranial height to cranial base length.The size of palatopharyngeal space(Ve-PVe) correlated negatively with the upper face height,percentage of cranial height to cranial base length.The size of hypopharyngeal space(Ph1-Ph2) correlated positively with the angle between cervical vertebrae tangent and cranial base, negatively mandibular plane angle, the length of mandibular body and percentage of cranial height to cranial base length. CONCLUSIONS: The sagittal upper airway size of non-snoring males with occlusion is significantly correlated to the craniofacial skeletal structures and head posture,the types of craniofacial growth and the skeletal structure around have remarkable effect on the upper airway space. PMID- 19290423 TI - [A sampled investigation on tooth loss of middle-aged and elders in Northeast China]. AB - PURPOSE: To investigate the current status and the distribution of tooth loss in middle-aged people and elders in Northeast China, and provide scientific basis for the establishment of oral health care policies. METHODS: An equal-sized stratified multi-stage random sampling design was applied according to the third national oral health investigation of epidemiology, tooth loss was examined among 4752 people aged 35-44 and 65-74 in northeast China. The data were analyzed with SPSS13.0 statistical software for independent-sample t test and Chi-square test. RESULTS: Of all the subjects, 3135 people had tooth loss, the rate of tooth loss was 66.80%, mean tooth loss was 4.95; 392 persons were half edentulous, the rate of edentulous jaw was 8.35%, 223 persons were edentulous, the rate edentulous jaw was 4.75%. Statistical analysis showed tooth loss rate in females was significantly higher than that in males(P<0.05). There was significantly difference of tooth loss between city and rural area (P<0.05). There was significant difference in the amount and rate of tooth loss between Jilin Province and Liaoning Province, and Jilin Province and Heilongjiang Province as well (P<0.05). CONCLUSIONS: The tooth loss rate is very high in middle-aged people and elders in Northeast China. Measures should be taken to prevent dental caries and periodontal disease in these population so that to promote the status of middle-aged people and elders on oral health and to improve their quality of life. PMID- 19290424 TI - [The relationship between familial oral hygiene behaviors and the caries risk factors in children]. AB - PURPOSE: The aim of this study is to explore the relationship between familial oral hygiene behaviors and caries risk factors of children and to speculate the possible influential mechanism in order to predict the onset of caries accurately and thoroughly and guide the oral hygiene behaviors accordingly as well. METHODS: The behavioral variables were acquired through delivering questionnaires and then the children were grouped based on the outcome of questionnaires; the caries risk factors were made necessary transformation; the relationship between them were analyzed with SPSS 12.0 software package for independent t test, stepwise logistic analysis, correlation and regression analysis. RESULTS: (1) Familial oral hygiene behaviors did influence the caries risk factors statistically. (2) Each kind of oral hygiene behavior had its special target caries risk factor and hence speculating its mechanism was different. (3)The oral hygiene behavior of children might be the result of a complicated influence of many factors including the parents, the teachers and other facets such as the media. CONCLUSION: Besides the conventional caries risk factors, the behavioral variables should be considered to be incorporated into the caries predict model. PMID- 19290425 TI - [Clinical evaluation of the effect of gold alloy and Ni-Cr alloy porcelain fused metal crown restorations]. AB - PURPOSE: To observe the clinical effect of gold alloy porcelain fused metal (PFM) crown restoration and Ni-Cr alloy PFM crown restoration. METHODS: A total of 168 teeth from 48 patients were restored with gold alloy PFM crown. The other 48 patients, with a total of 179 teeth were restored with Ni-Cr alloy PFM crown. They were examined in integrality, retention, shade, cervical margin, and gingival health immediately, 6 months, one year, two years ,and three years after restoration. The date was analyzed by rank sum test using SPSS12.0 software package. RESULTS: The clinical effect of Ni-Cr alloy PFM crown was as good as gold alloy PFM crown when checked up after cementation at once. However, when they were examined 6 months, one year, two years ,and three years after restoration, the clinical effect of gold alloy PFM crown group was significantly better than that of Ni-Cr alloy PFM crown, P<0.05. CONCLUSION: The gold alloy PFM crown has better properties than Ni-Cr alloy PFM crown as a kind of long-term restoration, especially on the aspect of shade. PMID- 19290426 TI - [Clinical study on glass fiber-reinforced post applied in the restoration of residual crown and root of posterior teeth]. AB - PURPOSE: The aim of this prospective randomized controlled trial is to evaluate the clinical effect of glass fiber-reinforced post core applied in the restoration of premolar or molar residual crown and root. METHODS: 195 residual crown and root of posterior teeth from 100 patients, aged from 18 to 60 years, were included in the study. Among 195 teeth, 95 teeth in the experimental group were restored with glass fiber-reinforced post core, while the other 100 teeth in the control group were restored with metal cast post core. All the teeth were then restored with porcelain fused metal crown. The esthetical property, function, and survival rate of the prostheses were observed. The data was analyzed with SPSS13.0 software package for X(2) test. RESULTS: The prostheses were followed up in this clinical study for a period of up to 36 months. In the experimental group, there were a total of 90 teeth with excellent chewing function, no complaint of discomfort, no debounding, no gingivitis or periapical periodontitis. The success rate of the experimental group was 94.74%, while that of the control group was 85.00%. The difference of the success rate between two groups was statistically significant(P<0.05). CONCLUSION: Glass fiber-reinforced post core can be used as an ideal restoration material for the premolar or molar residual crown and root. PMID- 19290427 TI - [Clinical observation of 45 cases with radicular cyst treated with non-surgical methods]. AB - PURPOSE: To observe the clinical effect and the influential factors of non surgical treatment of radicular cyst. METHODS: A total of 45 cases with radicular cyst were included from 2002 to 2008. They were treated by non-surgical treatment including decompression of cyst cavity or overfilling of the root canal. All the patients were examined at 3,6 months and 1,2,3 years after treatment. The therapeutic result was analyzed with SPSS15.0 software package for X(2) test. RESULTS: Of the 45 cases, 22 were cured at a follow-up period of 1-4 years, the cure rate was 48.89%. Seventeen cases were improved, the improvement rate 37.78%, and the total efficiency rate was 86.67%. Six cases failed to the treatment, and the failure rate was 13.33%. CONCLUSIONS: With decompression of cyst cavities or overfilling of the root canals, non-surgical treatment of radicular cyst is a simple, economical, safe, and effective treatment modality. PMID- 19290428 TI - [Jaw bone density assessments of implant sites using spiral CT and Simplant software]. AB - PURPOSE: To assess the bone density of dental implant sites using spiral CT and simplant software, and classify them by Lekholm and Zarb grouping. METHODS: 53 patients' jaw bone were scanned by spiral CT, the scanned images were reconstructed by the Simplant software.The bone density of 136 designed implant sites was measured and the results were recorded using Hounsfield units(HU).The data was analyzed with SPSS11.0 software package for Mann-Whitney U test, and classified by Lekholm and Zarb grouping. RESULTS: The mean bone density in all implant sites was (714.66+/-273.72) HU.The anterior mandible mean bone density was (962.96+/-92.21)HU>anterior maxilla, (786.15+/-188.74) HU>posterior mandible, (785.79+/-290.91) HU>posterior maxilla, (569.67+/-244.34)HU. Bone class IV was least in anterior mandible and most in posterior maxilla(P<0.01), accounting for 26.5% in all implant sites. CONCLUSIONS: Anterior mandible has the highest mean bone density and posterior maxilla has the lowest mean bone density. There are class I and II/III in most of the implant sites. Spiral CT is a useful tool for determining the bone density, which is important for preoperative design and postoperative assessment. PMID- 19290429 TI - [Mass chromatographic analysis on different protein expression of human periodontal ligament cell under static pressure]. AB - PURPOSE: To analyze the protein expression of human periodontal ligament cells (HPDLCs) under static pressure. METHODS: The proteins of two groups of human periodontal ligament cell (one group was interfered by static pressure and one group was not) were analyzed by two-dimensional electrophoresis and mass chromatographic analysis. The silver stained proteins spots were analyzed by Image Master 2D Platinum Software 5.0. There were about 720 and 730 detectable spots on the two 2D-gels separately and nearly 30 spots which were differentially expressed. RESULTS: With direct MALDI-TOF mass spectrometry and protein database searching, 2 protein spots out of 5 new appeared spots were identified. Presenilin 2 and catechol O-methyltransferase were expressed only in the group which was interfered by static pressure. CONCLUSIONS: The results of mass chromatographic analysis demonstrate that Notch pathway and calcitonin gene related peptide(CGRP) may be the partners of the channel of biomechanical signal conduction of the periodontal ligament cell. PMID- 19290430 TI - [Construction and expression of spap/A eukaryotic expression plasmid of Streptococcus mutans in mammalian cells]. AB - PURPOSE: The study is aimed to evaluate the expression of recombinant plasmid pVAX1-spap/A of surface protein antigen A of Streptococcus mutans in mammalian cells COS-7. METHODS: The eukaryotic plasmid carrying encoding gene of spap/A of Streptococcus mutans was constructed and the plasmid introduced into COS-7 cells by lipofectamine reagent. The transient expressed protein was detected by immunochemistry technique in COS-7 cells. RESULTS: Positive expression was detected in plasma of the cells which were transfected with recombinant plasmid pVAX1-spap/A. The cells which were transfected with pVAX1 were negative. CONCLUSIONS: Spap/A can translate and express in COS-7 cells after transfected with recombinant plasmid pVAX1-spap/A. The expressed protein locates in the plasma and the protein is able to combine with anti-spap/A antibody. The expressed protein has the antigenicity and recombinant plasmid pVAX1-spap/A is a candidate vaccine. PMID- 19290431 TI - [Study on the slow release of silver ion from silver containing antibacterial HA coating material]. AB - PURPOSE: To study the slow release of silver ion from silver containing antibacterial HA coating material. METHODS: Ti coated with HA samples were prepared using vacuum plasma spraying. In group 1, HA coating materials contained 5%(wt%) silver-zirconium phosphate antimicrobial. In Group 2, pure HA coating materials were used. The samples were immersed into the newborn calf serum and stored under anaerobic environment at constant temperatures of 37 degrees centigrade, avoiding light. The newborn calf serum was set as control. The Ag(+) ion concentration was detected and calculated using atomic absorption spectroscopy at 1,7,14 days. RESULTS: The Ag(+) ion contents of group 1 were 250 ng, 425 ng and 417 ng respectively at the end of 1,7,14 days. The release rate of Ag(+) ion became slow with the lapse of time. The release of Ag(+) ion became stable during 7-14 days. The Ag(+) ion content decreased on the 14th day. The Ag(+) ion content of group 2 could be ignored. CONCLUSIONS: The release of Ag(+) ion from silver containing HA coating materials is little and becomes stable on the 7th day. There might be reabsorption of Ag(+) ion on the 14th day. PMID- 19290432 TI - [Resistance to cyclic fatigue of pulpless teeth with flared root canals restored with three kinds of post-and-cores]. AB - PURPOSE: To observe the resistance to cyclic fatigue of maxillary incisors with flared canals restored with three post-and-core materials. METHODS: Standard cylindrical samples with flared canals were made of 48 human maxillary central incisors and were assigned randomly to two groups (without ferrule and 2mm ferrule). Each main group was then divided into 3 sub-groups of 8 specimens and restored with cast metal post-core-crown,carbon fiber+resin composite post-core crown ,and glass fiber+resin composite post-core-crown, respectively. Each group was subjected to a cyclic fatigue test, and the numbers of load cycles that had tooth fracture were measured with cyclic fatigue testing machine. All data were analyzed statistically for two-way ANOVA with SPSS 12.0 software package. RESULTS: Fiber+resin composite post-core-crown showed significantly higher fatigue strength than cast metal post-core-crown in the two groups(P<0.05). Preparing dentin ferrules increased significantly fatigue resistance(P<0.05). CONCLUSION: Fiber post-and-core system may get a long fatigue life in restoring pulpless teeth.Dentin ferrule preparation is necessary to enhance resistance to cyclic fatigue. PMID- 19290433 TI - [Effect of TiN coating on corrosion resistance of dental NiCr alloy]. AB - PURPOSE: To investigate the effect of titanium nitride coating on corrosion resistance of NiCr alloy in the simulate oral environment. METHODS: The NiCr alloy commonly used for porcelain crown was cast with specimen size of 10mm x 10mm x 3mm. Then the specimen was coated with a thickness of 2.5 microm titanium nitride coating on the surface by multi-arc physical vapor deposition. The Tafel polarization curves of the specimen before and after coating titanium nitride in artificial saliva were measured by electrochemical station. RESULTS: From the Tafel polarization curves, the specimen after coating titanium nitride showed that both anodic polarization curve and cathodal polarization curve were flat, and gained the higher natural corrosion potential, lower passivation maintaining current, longer passivation interval. The natural corrosion potential increased from -0.2487 V to -0.0685 V. The natural corrosion current density decreased from 0.2628 microA.cm(-2) to 0.1895 microA.cm(-2). The rupture potential increased from 0.8505 V to 1.2566 V. CONCLUSION: Titanium nitride coating can improve corrosion resistance of NiCr alloy. PMID- 19290434 TI - [3-D reconstruction of the maxillary central incisor by digital camera]. AB - PURPOSE: To evaluate the role of digital camera in reconstruction of 3-D solid model of the maxillary central incisor. METHODS: Photographs of each section of the maxillary central incisor were taken with digital camera, then the 3-D model was reconstructed with photoshop software and 3DMAX software. RESULTS: The precise 3-D solid model of the maxillary central incisor was successfully reconstructed. CONCLUSIONS: 3-D reconstruction of the maxillary central incisor can be completely accomplished by dentists themselves. PMID- 19290435 TI - [Effects of the micro-arc oxidized film on pure titanium surface on attachment and growth of BMSCs]. AB - PURPOSE: To evaluate the biocompatibility of oxide film formed by micro-arc oxidation(MAO) on titanium surface by culturing bone marrow stromal cells(BMSCs) on its surface and observing the BMSCs's biological behavior of early adhesion, growth and alkaline phosphatase activity. METHODS: According to the surface treatments, the samples were classified into 4 groups: MAO group, abrator group, abrator and NaOH-treated group, and untreated titanium group. Attachment, growth and alkaline phosphatase (ALP) activity of BMSCs cultured on the titanium surface of each group were assessed at different time. All data were statistically analyzed with one-way ANOVA using SPSS11.5 software package. RESULTS: From a morphological point of view, cell adherence in the MAO group was the best. Cell count and ALP activity in the MAO group were highest(P<0.01). CONCLUSIONS: MAO film on pure titanium surface may increase BMSCs adhesion, growth and ALP activity and titanium surface treated with MAO exhibits better biocompatibility. PMID- 19290436 TI - [Evaluation of BioPure MTAD as an endodontic irrigant on the apical sealing ability of AH Plus sealer in vitro]. AB - PURPOSE: To evaluate whether BioPure MTAD could affect the apical sealing ability of AH Plus sealer in vitro. METHODS: Forty-four extracted single-rooted human teeth were randomly divided into six groups. The canals were treated with distilled water,1.3%NaClO,5.25 %NaClO+17%EDTA,1.3%NaClO+BioPure MTAD and 1.3%NaClO+3%EDTA endodontic irrigants, respectively. After obturated, the teeth were made transparent and permitted dye penetration,then measured by using a stereoscopic microscope. SPSS 12.0 software package was used for One-way ANOVA. RESULTS: The microleakages lengths of obturated root canals in group C,D,E,F were (1.81+/-0.73)mm, (0.53+/-0.49)mm, (0.40+/-0.44)mm and (0.39+/-0.55)mm, respectively.Group C had significantly more leakage than group D,E,F(P<0.05), but there were no significant differences among group D,E,F(P>0.05). CONCLUSION: Compared with other endodontic irrigants which usually used in clinic, BioPure MTAD doesn't adversely affect the apical sealing ability of AH plus sealer. PMID- 19290437 TI - [Comparison of two means for evaluation of apical microleakage]. AB - PURPOSE: To compare the consistency of two means of evaluating the apical microleakage. METHODS: Forty-nine extracted single-rooted human teeth were divided into 3 experimental groups of 15 teeth each, in addition to a positive and negative control group of 2 teeth each. The teeth were instrumented, then obturated using lateral condensation and one of three endodontic sealers: AH Plus(group A), Cortisomol(group B) and zinc oxide eugenol(group C). The apical microleakage was measured by transparent tooth technique or dye dissolution each. SPSS 12.0 software package was used for One-way ANOVA and Kruslal-Wallis test. RESULTS: The two methods both showed that the microleakage in group A was significantly less (P<0.05) than that in group B and C. There was no significant difference between group B and C(P>0.05). CONCLUSION: Based on the dye penetration method, there is some consistency between the transparent tooth technique and dye dissolution method for evaluating apical microleakage. PMID- 19290438 TI - [Development of Chinese oral and maxillofacial surgery reflects the great achievements of China's reform and opening to the world for 30 years]. AB - China's policy of reform and opening to the world provides unlimited vitality for Chinese oral and maxillofacial surgery. The academic organization is increasingly perfecting, the professional team is expanding, and international exchanges are increasingly active. However, new challenges are emerging accordingly. To overcome the current problems and difficulties, some constructive beneficial recommendations were proposed and discussed in this paper. PMID- 19290439 TI - [Analysis and countermeasure of dental graduates' psychological state]. AB - PURPOSE: To study the dental graduates' psychological state in general dentistry and help teachers take countermeasure during clinical teaching. METHODS: 43 dental graduates were investigated in August 2008 through questionnaire, involving attitude toward career, communication in practice and so on. RESULTS: 46.51% of dental students felt optimistic about their occupation foreground, 72.09% of dental students wished be a dentist after graduation, 60.47% students felt indecisive in practice, 69.77% of students thought the preclinical theoretical study was partly useful in the practice, 65.12% of students could communicate with patients successfully. CONCLUSIONS: The dental graduates have ambivalence in profession ideal and actuality, such as conflict between work enthusiasm and work ability, communion need and obstacle. The teachers should try to intercommunicate with students,adopt corresponding psychotherapy individually,alleviate their stress,help them complete the dental practice well. PMID- 19290440 TI - [The neuromuscular toxicity and clinical application of adriamycin]. AB - Adriamycin (doxorubicin) is commonly used in the treatment of malignant tumours. In recent years,retrograde adriamycin sensory ganglionectomy has been used in treatment of trigeminal neuralgia and has obtained good therapeutic effect. The mechanism of action, particularly, of the toxic effects with different medication methods and choice of doses for muscle cells and nerve cells is still unclear. This article reviewed the mechanism and feature of the toxicity of adriamycin effects on these cells and its advance in experimental study. The damage of adriamycin was highly selective and self-limited. The different effects of adriamycin with different administration routes and doses is also described. Adriamycin shows great potentiality in treatment of trigeminal neuralgia, facial spasm and some other neuromuscular diseases. PMID- 19290441 TI - [Use of rigid internal fixation in treatment of oral and maxillofacial comminuted fracture]. AB - PURPOSE: To evaluate the usefulness of rigid internal fixation in management of oral and maxillofacial comminuted fracture. METHODS: From July 2004 to June 2007, 79 patients with oral and maxillofacial comminuted fractures (including 45 males, 34 females, aged from 9 to 69 years, 11 mandibular fractures, 31 ZMC fractures, 37 maxillary fractures) were treated with open reduction and rigid internal fixation. RESULTS: Facial profile in 79 cases and cross-bite in 77 cases were corrected satisfactorily. Cross-bite in 2 cases changed to open bite,and were corrected later with subsequent adjustment. CONCLUSION: Occlusal relation and facial contour can be corrected with rigid internal fixation in tension-line, stress-line or maxillofacial pillars with acceptable outcomes. PMID- 19290442 TI - [Clinical application of unimportant branches of facial nerve to repair important ones]. AB - PURPOSE: To study the feasibility and clinical effect of repairing important branches of facial nerve (zygomatic and marginal mandibular branches) with unimportant ones (upper buccal and cervical branches). METHODS: The defect of zygomatic and marginal mandibular branches was repaired with upper buccal and cervical branches, respectively. After the length of facial nerve defect was measured, the distal part of the upper buccal and cervical branches with peripheral tissue was used to repair the defect. Under operating microscope, nerve anastomosis was performed with 9-0 suture. 5 cases suffering from defect of marginal mandibular branches were repaired with cervical branches, and 7 cases with defect of zygomatic branches were repaired with upper buccal branches. RESULTS: In 5 cases suffering from defect of marginal mandibular branches, 2 cases recovered in 3 months, and 3 cases recovered in 6 months; In 7 cases suffering from defect of zygomatic branches, all recovered in 3-4 months. CONCLUSION: Repair of defect of zygomatic and marginal mandibular branches with upper buccal and cervical branches is feasible and the result is acceptable. PMID- 19290443 TI - [Investigation of indication for emergency surgical treatment of condylar fracture]. AB - PURPOSE: To investigate the indication for emergency surgical treatment of condylar fracture. METHODS: Eight patients with single condylar fracture and 3 patients with bilateral condylar fracture underwent emergency surgical treatment, the results were evaluated through radiographs and clinical examination. RESULTS: The mouth opening was normal with good occlusion,condylar anatomical contour was rehabilitated. CONCLUSIONS: Emergency surgical treatment of open condylar fracture is beneficial to healing of condylar fracture and reduction of occlusion. PMID- 19290444 TI - Detecting and predicting self-harm behaviour in prisoners: a prospective psychometric analysis of three instruments. AB - BACKGROUND: Research has revealed high levels of suicide and self-harm within young adult prisoners, but many studies have not considered the applicability and validity of its measurement for both male and female prisoners. Previous studies have focused on retrospective evaluations of instruments which are not useful evidence in informing clinical practice and decision making. OBJECTIVES: To evaluate the validation and prediction of suicide and self-harm risk in young adult prisoners. METHOD: The study was divided into two stages. Stage one used a cross-sectional design of 1,166 prisoners across six HM Prisons to validate the use of three questionnaires: the Beck Depression Inventory, the Beck Hopelessness Scale and a newly devised instrument (SCOPE tool). The second stage assessed the predictive validity of the three instruments using a 4-year-follow-up study of female prisoners across two HM Prisons in UK. Self-report and official records were used to measure suicide and self-harm risk. Logistic regression methodology, receiver operator characteristic curves and Youden's index were used to determine the range of thresholds for the three tools. RESULTS: Self-report measurement of suicide and self-harm behaviour using the three instruments presented a range of sensitivity and specificity values (65.9-72.3% and 64.9-74.0%, respectively). Predictive measurement of suicide and self-harm behaviour in the follow-up study presented a range of sensitivity and specificity values (54.6-80% and 62.2-69.4%, respectively). CONCLUSION: Screening for self-harm and suicidal behaviour in young prisoners has generated a range of cut off points for the identification of those at risk. These serve as a bench mark for service planners and practitioners. PMID- 19290445 TI - Effect of low concentrations of phosphate ions on extraction of arsenic from naturally contaminated soil. AB - The effect of low concentrations of phosphate ions on the extraction of arsenic (As) was tested on naturally contaminated soil (As 126 mg/kg) using batch experiments. Interaction entails the extraction of low concentrations of As (5.4 23.3 microg/L As). There are statistical differences in the contents of As, Ca, Cu, Ni, Pb and nitrate in the composition of the extracts. Three types of kinetic curves document that dissolution and precipitation reactions, adsorption, desorption and biological processes participate in determining the composition of the extracts. The elevated As contents in the phosphate extracts are probably caused by competition between phosphate and arsenate ions for soil adsorption sites. PMID- 19290446 TI - Distribution of toxic metals in organs of local cattle, sheep, goat and poultry from the West Bank, Palestinian Authority. AB - Graphite furnace atomic absorption spectrophotometry was used to measure the concentrations of Cu, Cd, Pb, and Zn in muscles and internal organs (liver, kidneys, heart, and lungs) of cattle, sheep goat and poultry sampled from local markets in the West Bank, Palestinian Authority. Mean levels of metals (dry weight basis) during this study were as follows: Cd: 0.34-0.57 microg/g, Pb: 0.2 4.7 microg/g, Cr: 0.44-3.62 microg/g and Cu: 1.03-217.9 microg/g. Muscles and internal organs of each species were found to have statistically different metal levels. Generally, livers and kidneys were found to have the highest levels of metals and muscles the lowest levels. When compared to each other, the four animal groups either did not show any significant differences or no clear trends. Compared to results reported in the literature, our results were comparable to or less than most of the studies reported from clean sites in different countries. PMID- 19290447 TI - First evidence of okadaic acid in Mytilus galloprovincialis mussels, collected in a Mediterranean Lagoon, Tunisia. AB - Evidence of the presence of okadaic acid (OA) and its monthly fluctuations are reported for the first time in Mytilus galloprovincialis samples collected from June 2005 to May 2006 in the Bizerte Lagoon. All of the samples with the exception of those taken in August 2005 were found to be contaminated. The level of OA in mussels exceeded the regulatory limit of DSP toxins (16 microg OA/100 g mussel meat) within the European Union (2002) in only two cases: in January and February, 2006 with 31.85 +/- 3.06, and 18.86 +/- 1.88 microg OA/100 g mussel meat respectively. During the investigative period, potentially toxic dinoflagellates were observed. The results reported here, although based on limited sampling, demonstrate for the first time the existence of a risk to public health from diarrheic shellfish poisoning in cultivated bivalve mollusc in Tunisia. PMID- 19290448 TI - Waste-indicator and pharmaceutical compounds in landfill-leachate-affected ground water near Elkhart, Indiana, 2000-2002. AB - Four wells downgradient from a landfill near Elkhart, Indiana were sampled during 2000-2002 to evaluate the presence of waste-indicator and pharmaceutical compounds in landfill-leachate-affected ground water. Compounds detected in leachate-affected ground water included detergent metabolites (p-nonylphenol, nonylphenol monoethoxylate, nonylphenol diethoxylate, and octylphenol monoethoxylate), plasticizers (ethanol-2-butoxy-phosphate and diethylphthalate), a plastic monomer (bisphenol A), disinfectants (1,4-dichlorobenzene and triclosan), an antioxidant (5-methyl-1H-benzotriazole), three fire-retardant compounds (tributylphosphate and tri(2-chloroethyl)phosphate, and tri(dichlorisopropyl)phosphate), and several pharmaceuticals and metabolites (acetaminophen, caffeine, cotinine, 1,7-dimethylxanthine, fluoxetine, and ibuprofen). Acetaminophen, caffeine, and cotinine detections confirm prior indications of pharmaceutical and nicotinate disposal in the landfill. PMID- 19290449 TI - Cadmium and other metal uptake by Lobelia chinensis and Solanum nigrum from contaminated soils. AB - Cadmium concentrations in two plant species and their corresponding soils were evaluated in a metal contaminated area. The average Cd concentrations reached 36.9 and 141 mg kg(-1) in Solanum nigrum leaves and Lobelia chinensis shoots, respectively. There is a significant relationship between the Cd concentration in the aerial tissues and the corresponding soils on a logarithmic scale. Under the hydroponic culture conditions, the maximum Cd concentration in the S. nigrum leaves and L. chinensis shoots were 1,110 and 414 mg kg(-1), respectively. Cd concentration was higher in the roots than in the aerial parts. The two plants may be used in suitable phytoremediation process. PMID- 19290450 TI - Quantitative study of Leydig cell populations in mice exposed to low doses of cadmium. AB - Leydig cell morphological changes were evaluated using morphometric and stereological methods in male mice exposed to low doses of cadmium. A possible reversibility of the changes after cadmium withdrawal was also considered. Nuclear morphological parameters and stereological densities of the Leydig cell population were lower in the cadmium-exposed groups than in the control. The withdrawal of cadmium did not lead to any significant recovery of the morphological parameters. Nevertheless, numerical density increased significantly in the withdrawn groups, suggesting that the hyperplasia of interstitial cells could try to relieve morphological damage after cadmium withdrawal. PMID- 19290451 TI - Distribution of phthalate esters in soil of e-waste recycling sites from Taizhou city in China. AB - In recent years, increasing concern has surrounded the consequences of improper electric and electronic waste (e-waste) disposal. In this paper, Phthalate esters (PAEs) including dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), Di-(2-ethylhexyl) phthalate (DEHP) and di-n-octyl phthalate (DnOP) in the e-waste soils were collected and analyzed from sites Fengjiang, Nanshan and Meishu in Taizhou city. The result showed that the total PAEs concentrations ranged from 12.566 to 46.669 mg/kg in these three sites. DEHP, DBP and DEP were the major phthalates accounting for more than 94% of total phthalates studied. Comparing to the results from other studies, the e-waste soils from Taizhou city were severely contaminated with PAEs. PMID- 19290452 TI - Degradation of the potato glycoalkaloids--alpha-solanine and alpha-chaconine in groundwater. AB - The potato glycoalkaloids alpha-chaconine and alpha-solanine are produced in high amounts in potato plants from where release to soil takes place. Degradation of the compounds in groundwater was investigated, as their fate in the terrestrial environment is unknown. Abiotic and microbial degradation were followed in groundwater sampled from below a potato field and spiked with the glycoalkaloids (115 nmol/l). Degradation was primarily microbial and the glycoalkaloids were degraded within 21-42 days. The metabolites beta(1)-solanine, gamma-solanine, and solanidine were formed from alpha-solanine, while beta-chaconine, gamma-chaconine and solanidine were detected from alpha-chaconine. Thus, indigenous groundwater microorganisms are capable of degrading the glycoalkaloids. PMID- 19290453 TI - Efficiency of experimental rice (Oryza sativa L.) fields in mitigating diazinon runoff toxicity to Hyalella azteca. AB - This study assessed the viability of using planted, mature rice fields in mitigating diazinon (an organophosphate insecticide) runoff toxicity using aqueous 48 h Hyalella azteca whole effluent toxicity bioassays. Rice fields decreased diazinon concentrations 80.1%-99.9% compared with 10.8% in the unvegetated field control. H. azteca survival responses coincided with observed diazinon concentrations. Estimated LC50 effects dilutions (%) ranged from 1.15 to 1.47 for inflow samples and 1.66 (unvegetated), 6.44 (rice field A), and >100 (rice field B) outflow samples. Decreases in inflow versus outflow aqueous toxicity were 77.1%-100% in rice fields compared with 18.7% in the unvegetated field. PMID- 19290454 TI - Daily variation of pesticides in surface water of a small river flowing through paddy field area. AB - The daily variations in the insecticides, i.e., buprofezin, etofenprox, fenitrothion, fenobucarb and pyridaphenthion, and fungicides, i.e., flutolanil, isoprothiolane, mepronil, pencycuron, phthalide and tricyclazole, were investigated in order to evaluate the short periodic variations in the pesticide concentrations of small river waters. The mean and maximum concentrations were 0.05 microg/L (pyridaphenthion)--1.2 microg/L (isoprothiolane) and 0.30 microg/L (pyridaphenthion)--13 microg/L (flutolanil), respectively. The concentration at 10:00 a.m. could be regarded as the representative daily concentration of the day. However, the concentration could markedly vary during the day or the day following the pesticide application. Therefore, the mean concentrations at 10 a.m., 2 p.m. and 6 p.m. were recommended as the best representative values of the day. PMID- 19290455 TI - Toxicity assessment of herbicides quizalafop-p-ethyl and clodinafop towards Rhizobium pea symbiosis. AB - In modern conventional agriculture, herbicides are frequently used to prevent yield losses due to weeds. Herbicides also affect negatively the productivity of legumes. With these considerations, we evaluated the effects of soil applications of different concentrations of quizalafop-p-ethyl and clodinafop on the performance of Rhizobium inoculated pea, grown in clay pots. In this study, the concentration of herbicides higher than the recommended rates of quizalafop-p ethyl and clodinafop adversely affected the dry matter accumulation, symbiotic properties, grain yield and nutrient status of pea plants. Toxicity of quizalafop p-ethyl and clodinafop to pea plants increased progressively with increase in rates of herbicides. Of the two herbicides, quizalafop-p-ethyl was more toxic than clodinafop. In contrast, when herbicide tolerant Rhizobium strain MRP1 was also used with herbicide, it increased the measured parameters at all concentrations. A maximum increase of 11%, 17%, 46%, 33%, 21% and 7% in the root N, shoot N, root P, shoot P, seed yield and seed protein, respectively, was observed when MRP1 was used with 120 microg quizalafop-p-ethyl kg(-1) soil while with 1,200 microg clodinafop kg(-1) soil it increased the root N, shoot N, root P, shoot P, seed yield and seed protein by 20%, 9%, 56%, 56%, 29% and 7%, respectively, compared with the un-inoculated but herbicide treated control. This study suggested that the toxic effects of herbicides on pea plants could be attenuated by applying growth promoting herbicide tolerant strain of Rhizobium under herbicide stressed soil environment. PMID- 19290456 TI - Quantitative health risk assessment of inhalation exposure to polycyclic aromatic hydrocarbons on citizens in Tianjin, China. AB - Considering the large amounts of PAHs emitted into the ambient air in China, it is urgent to take preliminary health risk assessment of citizens through inhalation exposure to PAHs in China. The incremental lifetime cancer risk (ILCR) model was used to get the risk level of Tianjin citizens as an example, and Monte Carlo simulation was adopted to deal with the uncertainty. Exposure analysis found that the average values of B[a]P equivalent (B[a]Peq) daily exposure doses for children in the indoor, traffic and outdoor settings were estimated to be 2,446.8, 478.4, and 321.6 ng day(-1), respectively. And those for adults were 3,344.1, 794.9, and 519.0 ng day(-1), respectively. Much attention must be paid to indoor exposure, as it contributes more than 70% of the B[a]Peq daily exposure dose. ILCR falls within the range of 10(-5)-10(-3), which is higher than the acceptable risk level of 10(-6), and lower than the priority risk level (10(-3)). So this risk should be compared with those of other public health issues in the purpose of risk management. Sensitivity analysis found that the two variables, indoor air PAHs concentration distribution and the cancer slope factor (CSF) of BaP, contribute about 89% of the total risk uncertainty. Thus they are considered as the two main factors influencing the accuracy of the PAHs health risk assessment. PMID- 19290457 TI - Regression of the level of different heavy metals to size of marine organisms harvested from the "Jiyeh" oil spill zone of the eastern Mediterranean Sea. AB - This study aimed to establish a baseline data on regression of the levels of Lead (Pb), Nickel (Ni) and Vanadium (V) to specific size dimensions of selected marine organisms harvested from an oil spill zone of the Eastern Mediterranean Sea. Twenty samples of each of Siganus rivulatus, Mullets and oysters were collected from each of six harvest sites. A total of 1, 3, and 3 respective significant regression equations (p < 0.01) were established between Pb, Ni, V and specific size dimensions of the selected marine organisms. The significant correlation associated with the highest R (2) value was obtained between the Pb (y) level and the width (x) of the Siganus rivulatus (y = -86.833x + 417.72). The other six statistically significant correlations were associated with lower values of R (2) ranging between 0.338 and 0.380. This baseline data will be used in the future to evaluate the self-purification process of pollutants in different sizes of indicator-marine organisms in this part of the Mediterranean Sea. PMID- 19290458 TI - Long-term (1996-2006) variation of nitrogen and phosphorus and their spatial distributions in Tianjin coastal seawater. AB - The spatio-temporal varying characteristics of dissolved inorganic nitrogen, reactive phosphate (RP), chemical oxygen demand (COD), and dissolving oxygen (DO) in Tianjin coastal seawater were investigated based on observation from May 1996 to October 2006. The concentrations of dissolved inorganic nitrogen (DIN), RP and COD ascended gradually and their varying ranges were 0.103-2.432, 0.009-0.12, and 0.8-2.9 mg L(-1), respectively. While DO in seawater decreased from 8.9 to 6.1 mg L(-1) gradually. Those indicated that human-induced eutrophication occurred and the seawater quality deteriorated. The spatial distributions of DIN, RP and COD were largely uniform, where isopleths generally descended from estuarine zones and bays to the central areas and from northern area to southern area, indicating that continental input is the dominant source of those pollutants. Especially, peak zones of those pollutants usually appeared near estuaries, Tianjin harbors, and dumping site of dredged sediment, which indicates that the urban and industrial sewage, shipping waste, dredged soil were the main sources for those contaminants in seawater. PMID- 19290459 TI - Beta2-integrin-mediated adhesion and intracellular Ca2+ release in human eosinophils. AB - Human eosinophils spontaneously adhere to various substrates in the absence of exogenously added activators. In the present study a method was developed for characterizing eosinophil adhesion by measuring changes in impedance. Impedance measurements were performed in HCO(3)-buffered HybriCare medium maintained in a humidified 5% CO(2) incubator at 37 degrees C. Impedance increased by more than 1 kOmega within minutes after eosinophils made contact with the substrate, reaching a peak within 20 min. Blocking mobilization of intracellular [Ca(2+)] that precedes adhesion with BAPTA-AM (10 microM) completely inhibited the rise in impedance as well as the changes in cell shape typically observed in adherent cells. However, lowering the extracellular [Ca(2+)] with 2.5 mM EGTA did not inhibit the increase in impedance. Pretreatment with anti-CD18 antibody to block substrate interactions with beta(2)-integrins, or jasplakinolide (2 microM) to block actin reorganization, abolished the increase in impedance and adherent morphology of the cells. Exposure of eosinophils to the phosphatidylinositol 3 kinase inhibitor LY294002 (5 microM) or treatment with protein kinase C zeta pseudosubstrate to competitively inhibit activity of the enzyme significantly reduced the increase in impedance and inhibited the cell spreading associated with adhesion. These results demonstrate a novel method for measuring eosinophil adhesion and showed that, following formation of a tethered attachment, a rapid increase in intracellular [Ca(2+)] precedes the cytoskeletal rearrangements required for cell shape changes and plasma membrane-substrate interactions associated with adhesion. PMID- 19290460 TI - Prevention of iatrogenic bile duct injuries in difficult laparoscopic cholecystectomies: is the naso-biliary drain the answer? AB - BACKGROUND: Prevention of iatrogenic injuries is of paramount importance in difficult laparoscopic cholecystectomies (LC). The objective of this study was to analyze the effectiveness of cholangiography using a pre-inserted endoscopic naso biliary drain (ENBD) for navigation during difficult cholecystectomies. METHODS: The study design was a retrospective case analysis. In 508 patients who underwent LC in a tertiary referral university hospital from 1996 through 2007, difficult cholecystectomy was anticipated in 26 patients due to possibly aberrant biliary anatomy (four patients), unclear cystic duct anatomy during magnetic resonance cholangiopancreatography (MRCP) and/or endoscopic retrograde cholangiopancreatography (ERCP) (three patients), and acute cholecystitis (19 patients). An ENBD was inserted during ERCP prior to LC for cholangiography (ENBDC) to facilitate safe dissection during LC. Prevalence of biliary complications was assessed as the main outcome measurement. RESULTS: The majority (68%) of the patients who underwent ENBDC had complicated cholecystitis. Advanced technical expertise was not required for insertion of an ENBD. In retrospect, ENBDC was useful in prevention of a possible catastrophe in 69% of cases. Open conversion was necessary in five patients and biliary complications occurred in five patients only in the non-ENBD group. There were no procedure-related complications. One limitation of the study was that it was not randomized and there was no comparison with patients without ENBDC. CONCLUSIONS: ENBDC is a useful and safe tool in the prevention of iatrogenic bile duct injuries in LC. PMID- 19290461 TI - Portal vein tumor thrombus from colorectal cancer with no definite metastatic nodules in liver parenchyma. AB - Portal vein tumor thrombus (PVTT) in hepatocellular carcinoma (HCC) is a common entity. In colorectal liver metastasis, microscopic tumor invasion into the intrahepatic portal vein is also usually observed, but the incidence of macroscopic tumor thrombus in the first branch and trunk of the portal vein is rare. Most reported cases of PVTT from colorectal cancer had concomitant metastatic nodules in liver parenchyma, and the PVTT was continuous with the liver nodule, like PVTT in HCC. We present a case of PVTT from colorectal cancer with no definite metastatic nodules in liver parenchyma. A 58-year old man underwent laparoscopic high anterior resection for rectosigmoid carcinoma accompanied by bulky tumor thrombus in the branch of the inferior mesenteric vein. Six months later, he received left lobectomy and left caudate resection for liver metastasis. The resected specimen demonstrated there was no metastatic nodule in liver parenchyma and that the left portal system was filled with the tumor thrombus. The patient is alive with no sign of recurrence 66 months after hepatectomy. Even if there is a macroscopic PVTT from colorectal cancer, a better prognosis may be expected when the tumor can be completely resected en-bloc by anatomic hepatectomy including PVTT. PMID- 19290462 TI - Failure to metformin and insulin secretagogue monotherapy: an observational cohort study. AB - The aim of the present cohort study is the assessment of treatment failure rates in patients on monotherapy with metformin or insulin secretagogues, observed in a routine clinical setting. A cohort of patients without any pharmacological treatment was also observed. A retrospective observational cohort study was performed on a consecutive series of 2,020 type 2 diabetic patients receiving monotherapy with an oral agent (metformin or insulin secretagogue, n = 1,126) or drug-naive (n = 894). HbA1c and prescribed hypoglycemic therapy were recorded yearly. Patients were followed until death, change of residence, failure to treatment, or up to 48 months. The mean duration of follow up was 34.8 +/- 18.0 months. In a Cox regression analysis, metformin was associated with a significant reduction, and insulin secretagogues with a significant increase, in the risk of failure to therapy during follow up. When duration of diabetes and baseline BMI were added to the model, insulin secretagogues, but not metformin, were still associated with increased risk of failure. In conclusion, insulin secretagogues are associated with increased failure rate in comparison with metformin. This difference could be due to detrimental effect of secretagogues, rather than to a beneficial action of metformin. PMID- 19290463 TI - Multicentric infantile myofibromatosis: a rare disorder of the calvarium. AB - INTRODUCTION: Infantile myofibromatosis is a rare mesenchymal disorder that occurs predominantly in infancy and early childhood, in either solitary or multicentric form. It can affect soft tissue, muscle, skeleton, and occasionally, visceral organs. Infantile myofibromatosis without visceral involvement frequently undergoes spontaneous regression. Multicentric infantile myofibromatosis with involvement exclusively of the calvarium is extremely rare. DISCUSSION: We report an 8-month-old girl who presented with multifocal calvarial lesions. The child underwent total excision of the temporal mass, and histopathological study gave a diagnosis of infantile myofibromatosis. Serial follow-up by neuroimaging was obtained at 3, 6, 12, and 24 months postoperatively. Three months after surgery, a new lesion in the midline of frontal bone was found, and there was partial regression of the occipital lesion. Complete regression of the untreated lesions was shown at 24 months. Illustrated by our patient and literature review, we emphasize the importance of recognition and proper intervention for this rare, nonmalignant disorder. PMID- 19290464 TI - Sylvian cistern dermoid cyst presenting with dysgeusia. AB - It is rare for a patient with a dermoid cyst (DC) to present with taste dysfunction. A 58-year-old man presented with an altered sense of taste. Magnetic resonance imaging of the brain showed a mass lesion in the right insula with compression effect. The tumour was resected and was found to be a DC. The postoperative course was uneventful, and the patient resumed his usual taste sensation within the early postoperative period. To the best of our knowledge, this is the first report of a sylvian cistern DC that presented with dysgeusia. PMID- 19290465 TI - Disseminated intravascular coagulation after isolated mild head injury. AB - A rare case is described of acute disseminated intravascular coagulation (DIC) following isolated mild head injury with acute subdural haematoma, coagulopathy onset preceding craniotomy. Surgical treatment of the cause followed by swift diagnosis and treatment soon after surgery enabled a good outcome. Post-operative recollection of subdural and extadural blood was treated by further surgery. DIC following isolated mild head injury without axonal damage is rare, but fatal if missed. Thrombocytopaenia in head injured patients should be investigated expediently. Post-operative interim imaging (if not standard practice) should also be considered to exclude haemorrhagic recollection requiring further surgery. PMID- 19290466 TI - Neurocryptococcosis in a 10-year-old immunocompetent girl. AB - We report an unusual case of a 10-year-old girl with neurocryptococcosis that was diagnosed only after histological examination of the cyst fluid obtained at surgery. PMID- 19290467 TI - Essential tremor leading to toxic liver damage successfully treated with deep brain stimulation. AB - PURPOSE: We present a patient with severe essential tremor (ET), who underwent thalamic deep brain stimulation (DBS). After previous medical treatment with Propranolol and Primidone failed, the patient resorted to alcohol, which greatly alleviated the symptoms. The downside of this situation, however, was that it led to alcoholism with severely disturbed liver enzymes and hepatic steatosis. METHODS & RESULTS: After DBS the patient was free of tremor and thus could easily abstain from alcohol. CONCLUSION: In time the elevated liver enzymes returned to normal, which indicates that sufficient tremor control by DBS can help these patients to abstain from alcohol. PMID- 19290468 TI - Intraventricular ganglioglioma: a short illustrated review. AB - The following review of the literature describes the ganglioglioma, an uncommon mixed glioneuronal neoplasm, most often of low-grade histology, with a small, albeit well-documented, malignant potential. These tumors exhibit a strong epileptogenic propensity and most often present as new onset seizures or are discovered after a long history of refractory epilepsy. Despite their indolent course, the importance of gross total resection is well recognized to prevent anaplastic and malignant degeneration. Morphologically, the neoplasm is often cystic with an enhancing mural nodule, but can also be entirely solid. They are most often found in the temporal lobe but have been found throughout the neuraxis. An exceedingly rare location of the ganglioglioma is within the lateral ventricle. A systematic literature search revealed only eight reports documenting the occurrence of a ganglioglioma within the lateral ventricle. We describe an illustrative case of an intraventricular ganglioglioma with a prominent cystic component and enhancing mural nodule, which represents the classic radiographic appearance of gangliogliomas described in other locations. A superior parietal lobule approach offered excellent surgical access for tumor removal and the patient has remained free of neurological deficits following surgery. Regardless of location within the central nervous system, ganglioglioma should be on the differential diagnosis for any cystic mass with a mural nodule, particularly in the setting of epilepsy. PMID- 19290469 TI - Comparison of experimental aneurysms embolized with second-generation embolic devices and platinum coils. AB - PURPOSE: The purpose of the study was to compare the performance of second generation embolic devices with that of platinum coils in experimental aneurysms. METHODS: Microsurgically constructed bifurcation aneurysms in rabbits were embolized with platinum coils (n = 7), HydroCoils 10 (n = 10), HydroSoft (n = 14) or Cerecyte (n = 6) devices. After 1 month, angiographic occlusion was scored and the aneurysms were histologically evaluated by light microscopy. Continuous and ordinal results were compared using ANOVA/Tukey-Kramer HSD and chi(2) tests respectively. RESULTS: Angiographic occlusion at follow-up was increased in the HydroCoil and HydroSoft groups and decreased in the platinum coil and Cerecyte groups. Fibrovascular tissue was observed in the sac of the Cerecyte group, while mixtures of fibrovascular tissue and fibrinous thrombus were observed in the other three groups. The inflammatory response and endothelialization of the neck were similar in all groups. CONCLUSIONS: Expansile hydrogel devices have led to increased progressive occlusion, while degradable polymer devices led to an increased rate of thrombus organization compared with platinum coils. PMID- 19290470 TI - Osteochondromas of the cervical spine in atypical location. AB - In the current article we review the available English literature by pubmed search engine on the topic of osteochondromas and their location in the cervical spine. The focus is to investigate the location of the lesions in cervical spine and to analyze clinical presentations and symptoms of patients, the aetiology and histopathology examination of the masses. In addition we report a rare illustrative case of a 62 year-old man with an osteochondroma of the cervical spine. The mass developed at a very slow rate for many years and produced no clinical symptoms. The location of the extradural mass in the right atlanto-axial joint of C1 and C2 is extremely rare and was not been reported so far. After preoperative CT- and MRI-imaging the entire mass could be removed. PMID- 19290471 TI - "Phosphene": early sign of vascular compression neuropathy of the optic nerve. AB - BACKGROUND: Phosphenes, flashes of light, are a visual phenomenon experienced by patients with ophthalmological disease and normal individuals. CASE REPORT: We report here a 68-year-old woman in whom phosphenes appeared in the left visual field due to compression of the right optic nerve by an aneurysm of the anterior communicating artery. RESULTS: The symptom decreased dramatically after clipping of the aneurysm. CONCLUSION: Phosphenes may be an important early sign of vascular compression neuropathy of the optic nerve. PMID- 19290472 TI - Hypoglossal schwannoma-successful reinnervation and functional recovery of the tongue following tumour removal and nerve grafting. AB - OBJECTIVE: Hypoglossal nerve schwannomas are rare tumours that usually cause ipsilateral hypoglossal palsy. This report describes such lesions in two patients and suggests nerve grafting as part of the treatment regimen. METHOD: Two patients with intra- and extra-dural hypoglossal schwannomas respectively were treated by direct surgery via a postero-lateral approach to the posterior fossa, hypoglossal canal and carotid sheath. Following tumour removal, sural nerve grafting was used to reconstruct the nerves. Unexpectedly, muscle bulk and motor function returned within 6 months in both patients. CONCLUSION: Nerve grafting was highly successful in achieving functional recovery following surgery for hypoglossal nerve schwannomas. PMID- 19290473 TI - Postoperative pain management after inguinal hernia repair: lornoxicam versus tramadol. AB - BACKGROUND: In this randomized and prospective study, we compared the analgesic effects of lornoxicam and tramadol in patients after inguinal hernia repair. METHODS: A total of 160 patients were assigned in a randomized manner into two groups. Group L received 8 mg lornoxicam i.v. at the end of the operation, followed by 8 mg 12 h after the operation. Group T received 1 mg/kg tramadol at the end of the operation and every 6 h up to 24 h postoperatively. The visual analog scale (VAS) score was assessed at 0, 2, 4, 8, 12, and 24 h after surgery. RESULTS: All patients completed the study. All vital signs were within normal ranges. The mean VAS score in Group L and in Group T was 21.66 +/- 14.64 and 19.75 +/- 11.82, respectively. No significant differences were found between groups with respect to VAS score. Eight (10%) patients in Group T had nausea. CONCLUSION: Lornoxicam 8 mg i.v. and b.i.d., tramadol 1 mg/kg at the end of the surgery and every 6 h up to 24 h after inguinal hernia repair provided rapid and effective analgesia and was well tolerated. PMID- 19290474 TI - For whom the bell tolls? DING proteins in health and disease. AB - DING proteins, identified mainly by their eponymous N-terminal sequences, are ubiquitous in living organisms. Amongst bacteria, they are common in pseudomonads, and have been characterised with respect to genetics and structure. They form part of a wider family of phosphate-binding proteins, with emerging roles in phosphate acquisition and pathogenicity. Many DING proteins have been isolated in eukaryotes, in which they have been associated with very diverse biological activities, often in the context of possible signalling roles. Disease states in which DING proteins have been implicated include rheumatoid arthritis, lithiasis, atherosclerosis, some tumours and tumour-associated cachexia, and bacterial and viral adherence. Complete genetic and structural characterisation of eukaryotic DING genes and proteins is still lacking, though the phosphate binding site seems to be conserved. Whether as bacterial proteins related to bacterial pathogenicity, or as eukaryotic components of biochemical signalling systems, DING proteins require further study. PMID- 19290475 TI - Cohesins: chromatin architects in chromosome segregation, control of gene expression and much more. AB - Cells have evolved to develop molecules and control mechanisms that guarantee correct chromosome segregation and ensure the proper distribution of genetic material to daughter cells. In this sense, the establishment, maintenance, and removal of sister chromatid cohesion is one of the most fascinating and dangerous processes in the life of a cell because errors in the control of these processes frequently lead to cell death or aneuploidy. The main protagonist in this mechanism is a four-protein complex denominated the cohesin complex. In the last 10 years, we have improved our understanding of the key players in the regulation of sister chromatid cohesion during cell division in mitosis and meiosis. The last 2 years have seen an increase in evidence showing that cohesins have important functions in non-dividing cells, revealing new, unexplored roles for these proteins in the control of gene expression, development, and other essential cell functions in mammals. PMID- 19290476 TI - Fructan and its relationship to abiotic stress tolerance in plants. AB - Numerous studies have been published that attempted to correlate fructan concentrations with freezing and drought tolerance. Studies investigating the effect of fructan on liposomes indicated that a direct interaction between membranes and fructan was possible. This new area of research began to move fructan and its association with stress beyond mere correlation by confirming that fructan has the capacity to stabilize membranes during drying by inserting at least part of the polysaccharide into the lipid headgroup region of the membrane. This helps prevent leakage when water is removed from the system either during freezing or drought. When plants were transformed with the ability to synthesize fructan, a concomitant increase in drought and/or freezing tolerance was confirmed. These experiments indicate that besides an indirect effect of supplying tissues with hexose sugars, fructan has a direct protective effect that can be demonstrated by both model systems and genetic transformation. PMID- 19290477 TI - Functions of actin in endocytosis. AB - Endocytosis is a fundamental eukaryotic process required for remodelling plasma membrane lipids and protein to ensure appropriate membrane composition. Increasing evidence from a number of cell types reveals that actin plays an active, and often essential, role at key endocytic stages. Much of our current mechanistic understanding of the endocytic process has come from studies in budding yeast and has been facilitated by yeast's genetic amenability and by technological advances in live cell imaging. While endocytosis in metazoans is likely to be subject to a greater array of regulatory signals, recent reports indicate that spatiotemporal aspects of vesicle formation requiring actin are likely to be conserved across eukaryotic evolution. In this review we focus on the 'modular' model of endocytosis in yeast before highlighting comparisons with other cell types. Our discussion is limited to endocytosis involving clathrin as other types of endocytosis have not been demonstrated in yeast. PMID- 19290478 TI - Shedding light on DARC: the role of the Duffy antigen/receptor for chemokines in inflammation, infection and malignancy. PMID- 19290479 TI - The investigation of synovial genomic targets of bucillamine with microarray technique. AB - OBJECTIVE: To identify the molecular mechanisms of bucillamine activity, global gene expression analysis and pathway analysis were conducted using IL-1 beta stimulated human fibroblast-like synovial cells (FLS). METHODS: Normal human FLS were treated with IL-1 beta in the presence or absence of 10 and 100 microM bucillamine for 6 h. Total RNA was extracted and global gene expression levels were detected using a 44 k human whole genome array. Data were analyzed using Ingenuity pathway analysis. RESULTS: Numerous pathways were activated by IL-1 beta stimulation. At both concentrations, bucillamine suppressed nine signal pathways stimulated by IL-1 beta. CONCLUSIONS: Bucillamine effectively inhibited fibroblast growth factor (FGF) signaling and tight junction signaling activated by IL-1 beta in FLS. Suppression of these signal pathways may correlate with the pharmacologic mechanisms of bucillamine. In particular, the suppression of FGF signaling by bucillamine is remarkable because the activation of FGF signaling may be involved in rheumatoid arthritis pathology. PMID- 19290480 TI - Expansion of CD14+CD16+ peripheral monocytes among patients with aseptic loosening. AB - OBJECTIVE AND DESIGN: In this study, we have investigated the relevance of peripheral blood inflammatory CD14(+)CD16(+) monocytes phenotype to patients with aseptic loosening (AL). MATERIAL AND TREATMENT: Immunophenotypes of monocytes were examined among patients with AL (n = 43), patients with mechanical loosening (ML, n = 30), patients with stable implant (SI, n = 16), and patients with osteoarthritis (OA, n = 17) using flow cytometry. METHODS: Immunological assay was used to measure TNF-alpha and IL-1 beta levels in both sera and culture media of implant wear stimulated CD14(+)CD16(+) and CD14(++)CD16(-) monocytes. Periprosthetic tissues were collected during surgery for histological assessment. RESULTS: The frequency of CD14(+)CD16(+) monocytes showed significant increase in AL patients than in ML, SI, and OA patients. A positive association was found between the subpopulation of CD14(+)CD16(+) monocytes and plasma TNF-alpha and IL 1 beta level in AL patients. Furthermore, a positive correlation existed between the subpopulation of CD14(+)CD16(+) monocytes and the total histopathology score. CONCLUSION: The results indicate that CD14(+)CD16(+) monocytes represent a sensitive marker for the disease activity of AL, and may serve as an effective prognostic index to identify total joint replacement recipients who are at increased risk for osteolysis and progression of AL. PMID- 19290481 TI - Pediatric brain tumors: mutations of two dioxygenases (hABH2 and hABH3) that directly repair alkylation damage. AB - Alkylating agents, commonly used for brain tumor therapy, induce DNA and RNA lesions that, if not repaired, drive cells to apoptosis. Thus, cellular mechanisms that are responsible for nucleic acid repair are possibly involved in drug resistance. This work analyzes hABH2 and hABH3, two human Fe(II)-dependent dioxygenases in pediatric brain tumors that are treated with alkylating agents. We analyzed 25 brain tumor samples for hABH2 and hABH3 mutations; a subset of samples was tested for quantitative expression with Real-Time PCR. Sequencing analysis showed two new mutations in two glioma patients, one of hABH2 coding sequence (I141 V) and the other of hABH3 (D189 N). The mutation at codon 189 falls in a crucial region of the protein. All subjects analyzed by Real-Time PCR showed an enhanced expression of the two genes, particularly of hABH2. This is the first study of hABH2 and hABH3 in pediatric brain tumors; further molecular investigations of their mutations and expression may help determine their role in response to chemotherapy. PMID- 19290482 TI - Primary leptomeningeal oligodendrogliomatosis. AB - Primary leptomeningeal oligodendrogliomas (PLOs) are rare intracranial malignancies where tumors grow in the subarachnoid space without an obvious connection to the brain or spinal cord parenchyma. Adding to the three previously reported cases of PLO with no parenchymal involvement we report a fourth case of the same in this paper in a 50-year-old woman presenting with unrelenting headaches. CT scan of her head revealed hydrocephalus and MRI revealed diffuse enhancement of her leptomeninges throughout her brain and spine, prominent over the basilar region. Biopsy obtained using a frameless stereotactic biopsy showed sharply defined cell borders, clear cytoplasm, and rounded nuclei consistent with an oligodendroglioma. Our case suggests that PLO can mimic diffuse forms of granulomatous meningitis and should be suspected in patients that clinically and radiographically present like granulomatous meningitis but without blood or CSF markers for the same. PMID- 19290483 TI - Shared and novel molecular responses of mandarin to drought. AB - Drought is the most important stress experienced by citrus crops. A citrus cDNA microarray of about 6.000 genes has been utilized to identify transcriptomic responses of mandarin to water stress. As observed in other plant species challenged with drought stress, key genes for lysine catabolism, proline and raffinose synthesis, hydrogen peroxide reduction, vacuolar malate transport, RCI2 proteolipids and defence proteins such as osmotin, dehydrins and heat-shock proteins are induced in mandarin. Also, some aquaporin genes are repressed. The osmolyte raffinose could be detected in stressed roots while the dehydrin COR15 protein only accumulated in stressed leaves but not in roots. Novel drought responses in mandarin include the induction of genes encoding a new miraculin isoform, chloroplast beta-carotene hydroxylase, oleoyl desaturase, ribosomal protein RPS13A and protein kinase CTR1. These results suggest that drought tolerance in citrus may benefit from inhibition of proteolysis, activation of zeaxanthin and linolenoyl synthesis, reinforcement of ribosomal structure and down-regulation of the ethylene response. PMID- 19290484 TI - [Report of activities of the Central Ethics Committee for Stem Cell Research. 6th report after implementation of the stem cell regulation for the period 1 December 2007 to 30 November 2008]. PMID- 19290485 TI - The dysregulation of the endocannabinoid system in diabesity-a tricky problem. AB - Endocannabinoids (ECs) are small lipid mediators that play a critical role in energy metabolism. Human studies have shown that the EC tone in peripheral tissues positively correlates with increased adiposity. Furthermore, pharmacological inhibition of EC signaling results in weight loss in humans. However, the mechanisms that cause the dysregulation of the EC system in obesity are not well-understood. Since the clinical utility of currently available EC blockers is severely limited due to their side effects like depression and suicidal ideation that are caused by central effects, it is important to delineate the role of central and peripheral effects of EC signaling in regulating glucose and lipid metabolism. PMID- 19290486 TI - Robotic versus laparoscopic low anterior resection of rectal cancer: short-term outcome of a prospective comparative study. AB - BACKGROUND: The aim of this study is to compare the short-term results between robotic-assisted low anterior resection (R-LAR), using the da Vinci Surgical System, and standard laparoscopic low anterior resection (L-LAR) in rectal cancer patients. METHODS: 113 patients were assigned to receive either R-LAR (n = 56) or L-LAR (n = 57) between April 2006 and September 2007. Patient characteristics, perioperative clinical results, complications, and pathologic details were compared between the groups. Moreover, macroscopic grading of the specimen was evaluated. RESULTS: Patient characteristics were not significantly different between the groups. The mean operation time was 190.1 +/- 45.0 min in the R-LAR group and 191.1 +/- 65.3 min in the L-LAR group (P = 0.924). The conversion rate was 0.0% in the R-LAR groups and 10.5% in the L-LAR group (P = 0.013). The serious complication rate was 5.4% in the R-LAR group and 19.3% in the L-LAR group (P = 0.025). The specimen quality was acceptable in both groups. However, the mesorectal grade was complete (n = 52) and nearly complete (n = 4) in the R LAR group and complete (n = 43), nearly complete (n = 12), and incomplete (n = 2) in the L-LAR group (P = 0.033). CONCLUSION: R-LAR was performed safely and effectively, using the da Vinci Surgical System. The use of the system resulted in acceptable perioperative outcomes compared to L-LAR. PMID- 19290487 TI - Lymphadenectomy for colon cancer: is there a consensus? PMID- 19290488 TI - Loss of RUNX3 expression correlates with differentiation, nodal metastasis, and poor prognosis of gastric cancer. AB - BACKGROUND: RUNX3 is a major growth regulator of gastric epithelial cells that is involved in gastric tumorigenesis in both humans and mice. In this study, we investigated the involvement of RUNX3 in tumor progression, and in the prognosis of human gastric cancer. METHODS: We analyzed the extent of RUNX3 protein expression by immunohistochemistry in 95 primary gastric adenocarcinomas, and correlated expression levels with clinicopathological parameters. We examined the effects of pFlag/RUNX3 on cell growth, apoptosis, and caspase-3 expression in AGS and SNU1 gastric cancer cell lines by colony-forming assay, terminal deoxynucleotidyl transferase (TdT)-mediate deoxyuridine triphosphatase (dUTP) nick-end labeling (TUNEL) assay, and Western blot analysis, respectively. The pFlag/RUNX3 effects on AGS invasion and migration potentials were also evaluated. RESULTS: RUNX3 expression was lost in 37 (39%) cases of gastric cancer. The expression of RUNX3 in diffuse- and mixed-type cancers was less frequent than expression in intestinal-type cancer (P < 0.001 and P = 0.001, respectively). In addition, the loss of RUNX3 expression was associated with lymph node metastasis (P = 0.02), and correlated with poor gastric cancer survival (P = 0.018). The growth of gastric cancer cells was suppressed after pFlag/RUNX3 transfection. The re-expression of RUNX3 resulted in the upregulation of caspase-3 and promoted apoptosis. Furthermore, Re-expression of RUNX3 induced significant inhibitions of AGS cell invasion and migration in vitro. CONCLUSIONS: This work shows that loss of RUNX3 expression is highly associated with lymph node metastasis and poor prognosis of gastric cancer. The re-expression of RUNX3 may induce apoptosis and inhibit the growth as well as invasion/migration of cancer cells. These results indicate that the targeting of the RUNX3 pathway could represent a potential modality for treating gastric cancer. PMID- 19290489 TI - Magnetic resonance imaging and contralateral prophylactic mastectomy: the "no mas" effect? PMID- 19290490 TI - Expression of P-aPKC-iota, E-cadherin, and beta-catenin related to invasion and metastasis in hepatocellular carcinoma. AB - OBJECTIVES: Atypical protein kinase C iota (aPKC-iota) and its associated intracellular molecules, E-cadherin and beta-catenin, are important for cell polarization in tumorigenesis and progression. Expression of aPKC-iota, P-aPKC iota (activated aPKC-iota), E-cadherin, and beta-catenin in hepatocellular carcinoma (HCC) was measured, and correlation with clinicopathological characteristics of HCC was analyzed. METHODS: Paraffin-embedded tumor tissue was obtained from patients with HCC after resection without preoperative radiotherapy or chemotherapy. Gene expression was detected by polymerase chain reaction (PCR), and protein expression was detected by immunohistochemistry and Western blot analysis. Expressions of aPKC-iota, P-aPKC-iota, E-cadherin, and beta-catenin were analyzed with relation to the clinicopathological data. RESULTS: The gene and protein expression of aPKC-iota are obviously higher in HCC tissues than that in peritumoral tissues and normal tissues by semiquantitative PCR and immunohistochemistry methods. Accumulation of aPKC-iota in HCC cytoplasm and nucleolus inhibited the later formation of belt-like adherens junctions (AJs) and/or tight junctions (TJs) in cell-cell contact. E-cadherin was reduced and accumulation of cytoplasm beta-catenin was increased in HCC. The expression of aPKC-iota was closely related to pathological differentiation, tumor size, invasion, and metastasis of HCC. CONCLUSION: Accumulation of cytoplasm aPKC-iota may reflect pathological differentiation, invasion, and metastasis potential of HCC. In this regard, our study on HCC revealed the potential usefulness of aPKC iota, E-cadherin, and beta-catenin as a prognostic marker, closely related to pathological differentiation, invasion, metastasis, and prognosis of HCC. PMID- 19290491 TI - Comparison of outcome of open and laparoscopic resection for stage II and stage III rectal cancer. AB - BACKGROUND: Laparoscopic resection for advanced rectal cancer has not been widely accepted, and there are only few studies with survival data. This study aimed to compare the survival of patients who underwent laparoscopic and open resection for stage II and III rectal cancer. MATERIALS AND METHODS: Consecutive patients (open resection: n = 310; laparoscopic resection: n = 111) who underwent curative resection for stage II and III rectal cancer from June 2000 to December 2006 were included. The operative details, postoperative complications, postoperative outcomes, and survival data were collected prospectively. Comparison was made between patients who had laparoscopic and open surgery. RESULTS: The age, gender, medical morbidity, types of operation, and American Society of Anesthesiologists (ASA) status were similar between the two groups. There was also no difference in the mortality, morbidity, and pathological staging. Laparoscopic resection was associated with significantly less blood loss and shorter hospital stay. With the median follow-up of 34 months, there was no difference in local recurrence rates. The 5-year actuarial survivals were 71.1% and 59.3% in the laparoscopic and open groups, respectively (P = .029). In the multivariate analysis, laparoscopic resection was one of the independent significant factors associated with better survival (P = .03, hazards ratio: 0.558, 95% confidence interval: 0.339-0.969). Other independent poor prognostic factors included lymph node metastasis, poor differentiation, perineural invasion, presence of postoperative complications, and no chemotherapy. CONCLUSIONS: Laparoscopic resection for locally advanced rectal cancer is associated with more favorable overall survival when compared with open resection. PMID- 19290492 TI - Better prognosis of T2 gastric cancer with preoperative diagnosis of early gastric cancer. AB - BACKGROUND: Gastrectomy with D2 lymph node dissection is the standard treatment for locally advanced gastric cancer in Japan. However, in patients with a preoperative diagnosis of early gastric cancer, gastrectomy with limited lymph node dissection is generally selected as a treatment option, despite a pathological diagnosis of advanced gastric cancer. The aim of the present study was to clarify the clinicopathological characteristics and feasibility of limited lymph node dissection in patients with clinically early, but pathologically advanced, gastric cancer. MATERIALS AND METHODS: Clinicopathological characteristics, including the incidence and susceptible sites for lymph node metastasis, were investigated in 1528 patients with a final diagnosis of T2 gastric cancer treated at the Cancer Institute Hospital. For these patients, the results were compared between two groups including 266 patients preoperatively diagnosed as early T1 gastric cancer (EpT2) and 1262 patients preoperative diagnosed as locally advanced gastric cancer (ApT2). RESULTS: The pathological stage was lower (P < .001), and the 5-year survival rate was better (91.0% vs. 73.1%, P < .001) for the EpT2 group compared with the ApT2 group. Moreover, the N stage was smaller in the EpT2 group than in the ApT2 group, and the extension of lymph node metastasis was limited in 261 out of 266 patients (98%). CONCLUSION: Patients with EpT2 gastric cancer had a better 5-year survival rate than the patients in the ApT2 group. Gastrectomy with limited lymph node dissection could be indicated for patients with EpT2 gastric cancer unless intraoperative finding indicated obvious lymph node metastasis. PMID- 19290493 TI - Synchronous colorectal liver metastases: triumph of prospective randomized trials over observational bias leads to paradigm shift. PMID- 19290494 TI - In silico identification of new genetic variations as potential risk factors for Alzheimer's disease in a microarray-oriented simulation. AB - Genomic and proteomic studies of neurodegenerative disorders require complementary approaches to integrate the massive amount of data generated in high throughput experimental procedures. We propose a Bioinformatics pipeline in which expression studies guide the selection of candidate genes that should be screened for potential new genetic variations from a public expressed site tags (ESTs) database. Motivated by the former interest of our group in genetic polymorphisms involved with the immune system, we selected five genes from a previous expression microarrays study of hippocampal cornu ammonis (CA1) area of Alzheimer's Disease subjects (AD). The CLCbio Workbench Combined version 3.6.2. was initially used to build ESTs and mRNA files retrieved respectively from the Goldenpath of University of California Santa Cruz (UCSC) and National Center for Biotechnology Information (NCBI) databases and latter to perform multiple batches of Smith-Waterman alignments. A total of 116 ESTs sequences were selected after proper stringent parameters were applied to the first set of mismatches. The annotation revealed various classes of variations, most of them deletions (176). Amongst this specific group, some were frameshift deletions (35) and the virtual translation of a few others (5) were predicted to induce no change other than a single aminoacid removal, with no subsequent repercussions at the protein sequence. In addition, the analysis identified transitions (three), transversions (52), synonymous (41), non-synonymous (12), and deletions in 36 ESTs located in Untranslated Regions -UTRs (Supplementary data). Deletions are often associated to major genetics syndromes with dysmorphic features. However, various recent studies show that common microdeletions might be highly associated with common neuropsychiatric disorders such as schizophrenia, autism, mental retardation, or even in various ethnicities, detected in whole genome sequencing experiments. A virtual validation confirmed that some of the variations identified were previously reported and confirmed in DNA samples, showing that this method is a feasible way to detect genetic variations that merit further exploration in AD genetic risk factor association studies. PMID- 19290495 TI - Bioreactors in tissue engineering: scientific challenges and clinical perspectives. AB - In this Chapter we discuss the role of bioreactors in the translational paradigm of Tissue Engineering approaches from basic research to streamlined tissue manufacturing. In particular, we will highlight their functions as: (1) Pragmatic tools for tissue engineers, making up for limitations of conventional manual and static techniques, enabling automation and allowing physical conditioning of the developing tissues; (2) 3D culture model systems, enabling us to recapitulate specific aspects of the actual in vivo milieu and, when properly integrated with computational modeling efforts and sensing and control techniques, to address challenging scientific questions; (3) Tissue manufacturing devices, implementing bioprocesses so as to support safe, standardized, scaleable, traceable and possibly cost-effective production of grafts for clinical use. We will provide evidences that fundamental knowledge gained through the use of well-defined and controlled bio-reactor systems at the research level will be essential to define, optimize, and moreover, streamline the key processes required for efficient manufacturing models. PMID- 19290496 TI - Bioreactor technology in cardiovascular tissue engineering. AB - Cardiovascular tissue engineering is a fast evolving field of biomedical science and technology to manufacture viable blood vessels, heart valves, myocardial substitutes and vascularised complex tissues. In consideration of the specific role of the haemodynamics of human circulation, bioreactors are a fundamental of this field. The development of perfusion bioreactor technology is a consequence of successes in extracorporeal circulation techniques, to provide an in vitro environment mimicking in vivo conditions. The bioreactor system should enable an automatic hydrodynamic regime control. Furthermore, the systematic studies regarding the cellular responses to various mechanical and biochemical cues guarantee the viability, bio-monitoring, testing, storage and transportation of the growing tissue.The basic principles of a bioreactor used for cardiovascular tissue engineering are summarised in this chapter. PMID- 19290497 TI - Bioreactors for guiding muscle tissue growth and development. AB - Muscle tissue bioreactors are devices which are employed to guide and monitor the development of engineered muscle tissue. These devices have a modern history that can be traced back more than a century, because the key elements of muscle tissue bioreactors have been studied for a very long time. These include barrier isolation and culture of cells, tissues and organs after isolation from a host organism; the provision of various stimuli intended to promote growth and maintain the muscle, such as electrical and mechanical stimulation; and the provision of a perfusate such as culture media or blood derived substances. An accurate appraisal of our current progress in the development of muscle bioreactors can only be made in the context of the history of this endeavor. Modern efforts tend to focus more upon the use of computer control and the application of mechanical strain as a stimulus, as well as substrate surface modifications to induce cellular organization at the early stages of culture of isolated muscle cells. PMID- 19290498 TI - Bioreactors for connective tissue engineering: design and monitoring innovations. AB - The challenges for the tissue engineering of connective tissue lie in creating off-the-shelf tissue constructs which are capable of providing organs for transplantation. These strategies aim to grow a complex tissue with the appropriate mechanical integrity necessary for functional load bearing. Monolayer culture systems lack correlation with the in vivo environment and the naturally occur ring cell phenotypes. Part of the development of more recent models is to create growth environments or bioreactors which enable three-dimensional culture. Evidence suggests that in order to grow functional load-bearing tissues in a bioreactor, the cells must experience mechanical loading stimuli similar to that experienced in vivo which sets out the requirements for mechanical loading bioreactors. An essential part of developing new bioreactors for tissue growth is identifying ways of routinely and continuously measuring neo-tissue formation and in order to fully identify the successful generation of a tissue implant, the appropriate on-line monitoring must be developed. New technologies are being developed to advance our efforts to grow tissue ex vivo. The bioreactor is a critical part of these developments in supporting growth of biological implants and combining this with new advances in the detection of tissue formation allows us to refine our protocols and move nearer to off-the-shelf implants for clinical applications. PMID- 19290499 TI - Mechanical strain using 2D and 3D bioreactors induces osteogenesis: implications for bone tissue engineering. AB - Fracture healing is a complicated process involving many growth factors, cells, and physical forces. In cases, where natural healing is not able, efforts have to be undertaken to improve healing. For this purpose, tissue engineering may be an option. In order to stimulate cells to form a bone tissue several factors are needed: cells, scaffold, and growth factors. Stem cells derived from bone marrow or adipose tissues are the most useful in this regard. The differentiation of the cells can be accelerated using mechanical stimulation. The first part of this chapter describes the influence of longitudinal strain application. The second part uses a sophisticated approach with stem cells on a newly developed biomaterial (Sponceram) in a rotating bed bioreactor with the administration of bone morphogenetic protein-2. It is shown that such an approach is able to produce bone tissue constructs. This may lead to production of larger constructs that can be used in clinical applications. PMID- 19290500 TI - Bioreactors for tissue engineering of cartilage. AB - The cartilage regenerative medicine field has evolved during the last decades. The first-generation technology, autologous chondrocyte transplantation (ACT) involved the transplantation of in vitro expanded chondrocytes to cartilage defects. The second generation involves the seeding of chondrocytes in a three dimensional scaffold. The technique has several potential advantages such as the ability of arthroscopic implantation, in vitro pre-differentiation of cells and implant stability among others (Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L, N Engl J Med 331(14):889-895, 1994; Henderson I, Francisco R, Oakes B, Cameron J, Knee 12(3):209-216, 2005; Peterson L, Minas T, Brittberg M, Nilsson A, Sjogren-Jansson E, Lindahl A, Clin Orthop (374):212-234, 2000; Nagel-Heyer S, Goepfert C, Feyerabend F, Petersen JP, Adamietz P, Meenen NM, et al. Bioprocess Biosyst Eng 27(4):273-280, 2005; Portner R, Nagel-Heyer S, Goepfert C, Adamietz P, Meenen NM, J Biosci Bioeng 100(3):235-245, 2005; Nagel Heyer S, Goepfert C, Adamietz P, Meenen NM, Portner R, J Biotechnol 121(4):486 497, 2006; Heyland J, Wiegandt K, Goepfert C, Nagel-Heyer S, Ilinich E, Schumacher U, et al. Biotechnol Lett 28(20):1641-1648, 2006). The nutritional requirements of cells that are synthesizing extra-cellular matrix increase along the differentiation process. The mass transfer must be increased according to the tissue properties. Bioreactors represent an attractive tool to accelerate the biochemical and mechanical properties of the engineered tissues providing adequate mass transfer and physical stimuli. Different reactor systems have been [5] developed during the last decades based on different physical stimulation concepts. Static and dynamic compression, confined and nonconfined compression based reactors have been described in this review. Perfusion systems represent an attractive way of culturing constructs under dynamic conditions. Several groups showed increased matrix production using confined and unconfirmed systems. Development of automatic culture systems and noninvasive monitoring of matrix production will take place during the next few years in order to improve the cost affectivity of tissue-engineered products. PMID- 19290501 TI - Technical strategies to improve tissue engineering of cartilage-carrier constructs. AB - Technical aspects play an important role in tissue engineering. Especially an improved design of bioreactors is crucial for cultivation of artificial three dimensional tissues in vitro. Here formation of cartilage-carrier-constructs is used to demonstrate that the quality of the tissue can be significantly improved by using optimized culture conditions (oxygen concentration, growth factor combination) as well as special bioreactor techniques to induce fluid-dynamic, hydrostatic or mechanical load during generation of cartilage. PMID- 19290502 TI - Application of disposable bag bioreactors in tissue engineering and for the production of therapeutic agents. AB - In order to increase process efficiency, many pharmaceutical and biotechnology companies have introduced disposable bag technology over the last 10 years. Because this technology also greatly reduces the risk of cross-contamination, disposable bags are preferred in applications in which an absolute or improved process safety is a necessity, namely the production of functional tissue for implantation (tissue engineering), the production of human cells for the treatment of cancer and immune system diseases (cellular therapy), the production of viruses for gene therapies, the production of therapeutic proteins, and veterinary as well as human vaccines.Bioreactors with a pre-sterile cultivation bag made of plastic material are currently used in both development and manufacturing processes primarily operating with animal and human cells at small- and middle-volume scale. Because of their scalability, hydrodynamic expertise and the convincing results of oxygen transport efficiency studies, wave-mixed bioreactors are the most used, together with stirred bag bioreactors and static bags, which have the longest tradition.Starting with a general overview of disposable bag bioreactors and their main applications, this chapter summarizes the working principles and engineering aspects of bag bioreactors suitable for cell expansion, formation of functional tissue and production of therapeutic agents. Furthermore, results from selected cultivation studies are presented and discussed. PMID- 19290503 TI - Methodology for optimal in vitro cell expansion in tissue engineering. AB - Expansion of the cell population in vitro has become an essential step in the process of tissue engineering and also the systematic optimization of culture conditions is now a fundamental problem that needs to be addressed. Herein, we provide a rational methodology for searching culture conditions that optimize the acquisition of large quantities of cells following a sequential expansion process. In particular, the analysis of both seeding density and passage length was considered crucial, and their correct selection should be taken as a requisite to establish culture conditions for monolayer systems. This methodology also introduces additional considerations concerning the running cost of the expansion process. The selection of culture conditions will be a compromise between optimal cell expansion and acceptable running cost. This compromise will normally translate into an increase of passage length further away from the optimal value dictated by the growth kinetic of the cells. Finally, the importance of incorporating functional assays to validate the phenotypical and functional characteristics of the expanded cells has been highlighted. The optimization approach presented will contribute to the development of feasible large scale expansion of cells required by the tissue engineering industry. PMID- 19290504 TI - Bioreactor studies and computational fluid dynamics. AB - The hydrodynamic environment "created" by bioreactors for the culture of a tissue engineered construct (TEC) is known to influence cell migration, proliferation and extra cellular matrix production. However, tissue engineers have looked at bioreactors as black boxes within which TECs are cultured mainly by trial and error, as the complex relationship between the hydrodynamic environment and tissue properties remains elusive, yet is critical to the production of clinically useful tissues. It is well known in the chemical and biotechnology field that a more detailed description of fluid mechanics and nutrient transport within process equipment can be achieved via the use of computational fluid dynamics (CFD) technology. Hence, the coupling of experimental methods and computational simulations forms a synergistic relationship that can potentially yield greater and yet, more cohesive data sets for bioreactor studies. This review aims at discussing the rationale of using CFD in bioreactor studies related to tissue engineering, as fluid flow processes and phenomena have direct implications on cellular response such as migration and/or proliferation. We conclude that CFD should be seen by tissue engineers as an invaluable tool allowing us to analyze and visualize the impact of fluidic forces and stresses on cells and TECs. PMID- 19290505 TI - Fluid dynamics in bioreactor design: considerations for the theoretical and practical approach. AB - The following chapter summarizes principles of fluid dynamics in bioreactor design with a focus on mammalian cell-culture systems. PMID- 19290506 TI - [Is the structure of surgical clinics in Germany changing? A current investigation into the structure of surgical clinics in the Federal Republic of Germany]. AB - Medical developments have led to extensive specialization in the field of surgery. This has already been reflected for many years in altered structure and organization forms of surgical clinics. Indispensable quality standards, statutory general conditions, increasing competition in service providers and health insurance with transparency of the service procedure all intensify this trend. The aim of this investigation was, therefore, to determine how far this differentiation of service supply in the field of surgery is also reflected in the area and in surgical departments and clinics of basic and routine supply. To achieve this, all available published information on the structure and organization of surgical clinics in the Federal Republic of Germany was classified according to current departmentalization into "undivided" or general/visceral surgery facilities compared to orthopedic/trauma surgery departments. PMID- 19290507 TI - Soft tissue balancing in varus total knee arthroplasty: an algorithmic approach. AB - We present an algorithmic release approach to the varus knee, including a novel pie crust release technique of the superficial MCL, in 359 total knee arthroplasty patients and report the clinical and radiological outcome. Medio lateral stability was evaluated as normal in 97% of group 0 (deep MCL), 95% of group 1 (pie crust superficial MCL) and 83% of group 2 (distal superficial MCL). The mean preoperative hip-knee angle was 174.0, 172.1, and 169.5 and was corrected postoperatively to 179.1, 179.2, and 177.6 for groups 0, 1, and 2, respectively. A satisfactory correction in the coronal plane was achieved in 82.9% of all-comers falling within the 180 degrees +/- 3 degrees interval. An algorithmic release approach can be beneficial for soft tissue balancing. In all patients, the deep medial collateral ligament should be released and otseophytes removed. The novel pie crust technique of the superficial MCL is safe, efficient and reliable, provided a medial release of 6-8 mm or less is required. The release of the superficial MCL on the distal tibia is advocated in severe varus knees. Preoperative coronal alignment is an important predictor for the release technique, but should be combined with other parameters such as reducibility of the deformity and the obtained gap asymmetry. PMID- 19290509 TI - Chromium uptake, retention and reduction in photosynthetic Euglena gracilis. AB - Photosynthetic Euglena gracilis grown with different K(2)CrO(4) concentrations was analyzed for its ability to take up, retain and reduce Cr(VI). For comparison, cells were also exposed to CrCl(3). Cellular Cr(VI) uptake at pH 7.2 showed a hyperbolic saturation pattern with K (m) of 1.1 mM, V (m) of 16 nmol (h x 10(7) cells)(-1), and K (i sulfate) of 0.4 mM. Kinetic parameters for sulfate uptake were similar, K (m) = 0.83 mM, V (m) = 15.9 nmol (h x 10(7)cells)(-1) and K (i chromate) = 0.3 mM. The capacity to accumulate chromium depended on the ionic species, external concentration and pH of the incubation medium. Cr(VI) or Cr(III) accumulation was negligible in the acidic (pH 3.5) culture medium, in which Cr(VI) was abiotically reduced to Cr(III). At pH 7.2 Cr(VI) was fully stable and high accumulation (>170 nmol/1 x 10(7) cells at 1 mM K(2)CrO(4)) was achieved; surprisingly, Cr(III) accumulation was also significant (>35 nmol/1 x 10(7) cells at 1 mM CrCl(3)). Cr(VI) was reduced by cells at pH 7.2, suggesting the presence of an external reductive activity. Cr(VI) induced an increased cysteine and glutathione content, but not in phytochelatins suggesting that chromium accumulation was mediated by monothiol compounds. PMID- 19290508 TI - Anatomic double-bundle versus single-bundle ACL reconstruction: a comparative biomechanical study in rabbits. AB - Thirty New Zealand white rabbits underwent anterior cruciate ligament (ACL) reconstruction in their right knees; 15 animals underwent a double-bundle anatomic ACL reconstruction using the medial third of the patellar tendon and the semitendinosus tendon. Additionally, 15 animals underwent ACL reconstruction, using a single-bundle semitendinosus tendon autograft. The knees of both groups were evaluated with a device similar to the KT1000 arthrometer onto which a dial indicator was attached (Mitutoyo dial indicator 2050) in 30 degrees and 90 degrees of flexion, preoperatively, after ACL resection and 3 months postoperatively. Statistical analysis of the results revealed that for 90 degrees of knee flexion, the mean estimated anterior shift for the double-bundle technique was 1.92 mm lesser than that of the single-bundle technique (P = 0.006). For 30 degrees of knee flexion, the mean anterior shift was again lesser than that of the single-bundle technique by 0.66 mm, but this difference was not statistically significant. The described double-bundle ACL reconstruction technique resulted in a more stable knee as far as the anterior tibial shift was concerned as compared to a single-bundle ACL reconstruction. This animal model may be potentially useful in the future for the study of other parameters influencing the outcome of the double-bundle ACL reconstruction. PMID- 19290510 TI - Use of polyclonal antibodies to ochratoxin A with a quartz-crystal microbalance for developing real-time mycotoxin piezoelectric immunosensors. AB - A piezoelectric immunosensor was tested for ochratoxin A (OTA) mycotoxin detection through the immobilization of OTA-bovine serum albumin (OTA-BSA) conjugate on gold-coated quartz crystals (AT-cut/5 MHz). Immunoassays were performed in a flow-injection system through frequency decreases in a quartz crystal microbalance (QCM) because of a mass increasing during immunoreaction with anti-OTA antibodies. Three immobilization procedures for OTA-BSA (direct adsorption and covalent attachment to two alkane thiol self-assembled monolayers) were characterized with QCM in real time. Covalent attachment of the OTA-BSA conjugates through gold nanoparticles was also tested for amplifying the signal. Binding of the excess of antibodies to the immobilized OTA in an indirect competitive analysis decreased linearly the resonant frequency in the range of the OTA concentration from 10 to 128 ng/mL, with a detection limit of 8 ng/mL (signal/noise ratio of 3). A pepsin 2 mg/mL (pH = 2.1) solution was used to release antigen-antibody complexes, regenerating the biorecognition surface. PMID- 19290511 TI - Miniaturized 96-well ELISA chips for staphylococcal enterotoxin B detection using portable colorimetric detector. AB - A previously developed fluorescence sensing platform, combining spatial illumination using electroluminescence (EL) semiconductor strips with charge coupled device (CCD)-based detection (EL-CCD), was adapted to a new 96-well chip for colorimetric immunological assays, enhancing the capabilities of the EL-CCD platform. The modified system was demonstrated using a colorimetric-based enzyme linked immunosorbent assay (ELISA) for detection of staphylococcal enterotoxin B (SEB). Limits of detection (LODs) of 3.9 ng/mL (+/-2.4 ng/mL) SEB were determined with the ELISA chip measured using the EL-CCD platform, following a standard 4-h ELISA protocol. The LODs were comparable to those obtained using standard 96-well ELISA plates measured using a standard laboratory 96-well plate reader. The miniature 96-well ELISA chip however required as little as 5-microL samples, representing a tenfold reduction in sample volume compared to a standard 96-well ELISA plates. The ELISA chip also demonstrated detection of SEB spiked into various food matrices (milk, mushrooms, and mayonnaise) using limited-to-no sample preparation, with LODs ranging from 3.9 to 18.5 ng/mL depending on the matrix. The EL-CCD platform is versatile, capable of multi-mode detection (e.g., fluorescent and colorimetric along with solution and solid phase assays), and could readily be applied to other field portable or point-of-care applications. PMID- 19290512 TI - Halogenated molecularly imprinted polymers for selective determination of carbaryl by phosphorescence measurements. AB - A highly selective molecularly imprinted polymer (MIP) for the recognition of the pesticide carbaryl in water has been synthesized using halogenated bisphenol A compounds as one of the polymeric precursors and carbaryl as the template molecule. On the basis of the heavy-atom effect, both the brominated and the iodinated MIPs allowed analyte detection by room-temperature-phosphorescence measurements. In the presence of an oxygen scavenger (sodium sulphite) the halide, included in the polymeric structure, induced efficient room-temperature phosphorescence of the analyte (once it had been selectively retained by the MIP). The MIP cavity can be easily regenerated for subsequent sample injections with 2 mL methanol. The optosensing system developed has demonstrated high selectivity for carbaryl, even in the presence of other luminophores that could be unspecifically adsorbed onto the MIP surface. Under optimal experimental conditions, the detection limit for the target molecule was 4 microg/L (3-mL sample injection volume), and the linear range extended up to 1 mg/L of the analyte. Good reproducibility was achieved (a relative standard deviation of 3% was obtained for ten replicates of 150 microg/L carbaryl). The synthesized sensing material showed good stability for at least 3 months after preparation. Finally, the applicability to carbaryl determination in real samples was evaluated through the successful determination of the pesticide in spiked mineral and tap water samples. PMID- 19290513 TI - Hybrid gravitational field-flow fractionation using immunofunctionalized walls for integrated bioanalytical devices. AB - In this work, the biospecific recognition antigen-antibody reaction was implemented in gravitational field-flow fractionation (GrFFF), a flow-assisted separation technique for micron-sized particles, in order to realize a hybrid immunomodulated GrFFF system in which two different principles are combined to achieve highly versatile fractionation. Micron-sized polystyrene beads coated with horseradish peroxidase (HRP) were used as a model sample, and anti-HRP antibodies were immobilized on the accumulation wall of the GrFFF channel. Ultrasensitive chemiluminescence imaging was employed to visualize the beads during elution and to optimize experimental conditions. The same principle was then applied to real biological samples composed by Yersinia enterocolitica and Escherichia coli cells. Results show the possibility to modify the elution of selected sample components and even to retain them into the channel. The hybrid immunomodulated GrFFF system is a step towards the development of a module that could be integrated in a lab-on-a-chip-based point-of-care testing device which includes sample pre-analytical cleanup and analysis. PMID- 19290514 TI - Micro free-flow electrophoresis: theory and applications. AB - Free-flow electrophoresis (FFE) is a technique that performs an electrophoretic separation on a continuous stream of analyte as it flows through a planar flow channel. The electric field is applied perpendicularly to the flow to deflect analytes laterally according to their mobility as they flow through the separation channel. Miniaturization of FFE (microFFE) over the past 15 years has allowed analytical and preparative separation of small volume samples. Advances in chip design have improved separations by reducing interference from bubbles generated by electrolysis. Mechanisms of band broadening have been examined theoretically and experimentally to improve resolution in microFFE. Separations using various modes such as zone electrophoresis, isoelectric focusing, isotachophoresis, and field-step electrophoresis have been demonstrated. PMID- 19290515 TI - Damage to superior parietal cortex impairs pointing in the sagittal plane. AB - Neurophysiology and neuroimaging research implicates distinct regions of posterior parietal cortex for reaching versus grasping and for completing these movements in central versus peripheral space. Typically, visuomotor tasks only examine movements made in the frontoparallel plane. We examined a patient with a right superior parietal lesion encompassing the parietal-occipital junction, the intraparietal sulcus and the putative human homologue of V6A on pointing tasks in the sagittal or frontoparallel planes. The patient did not demonstrate a speed accuracy trade-off, but did show larger times post-peak velocity for all movement directions. Her movements in the sagittal axis were more disordered than movements in the frontoparallel plane. These data indicate a role for superior parietal cortex in fine tuning of visually guided movements and more particularly for movements made back towards the body. PMID- 19290517 TI - Solution structure of the major (Spy0128) and minor (Spy0125 and Spy0130) pili subunits from Streptococcus pyogenes. AB - Adhesion of the serotype M1 Streptococcus pyogenes strain SF370 to human tonsil explants and cultured keratinocytes requires extended polymeric surface structures called pili. In this important human pathogen, pili are assembled from three protein subunits: Spy0125, Spy0128 and Spy0130 through the action of sortase enzymes. For this study, the structural properties of these pili proteins have been investigated in solution. Spy0125 and Spy0128 display characteristics of globular, folded proteins. Circular dichroism suggests a largely beta-sheet composition for Spy0128 and Spy0125; Spy0130 appears to contain little secondary structure. Each of the proteins adopts a monodisperse, monomeric state in solution as assessed by analytical ultracentrifugation. Further, small-angle X ray scattering curves for Spy0125, Spy0128 and Spy0130 suggest each protein adopts an elongated shape, likely comprised of two domains, with similar maximal dimensions. Based on the scattering data, dummy atom models of each of the pili subunits have been reconstructed ab initio. This study provides the first insights into the structure of Streptococcus pyogenes minor pili subunits, and possible implications for protein function are discussed. PMID- 19290516 TI - Mode of action of cytokines on nociceptive neurons. AB - Cytokines are pluripotent soluble proteins secreted by immune and glial cells and are key elements in the induction and maintenance of pain. They are categorized as pro-inflammatory cytokines, which are mostly algesic, and anti-inflammatory cytokines, which have analgesic properties. Progress has been made in understanding the mechanisms underlying the action of cytokines in pain. To date, several direct and indirect pathways are known that link cytokines with nociception or hyperalgesia. Cytokines may act via specific cytokine receptors inducing downstream signal transduction cascades, which then modulate the function of other receptors like the ionotropic glutamate receptor, the transient vanilloid receptors, or sodium channels. This receptor activation, either through amplification of the inflammatory reaction, or through direct modulation of ion channel currents, then results in pain sensation. Following up on results from animal experiments, cytokine profiles have recently been investigated in human pain states. An imbalance of pro- and anti-inflammatory cytokine expression may be of importance for individual pain susceptibility. Individual cytokine profiles may be of diagnostic importance in chronic pain states, and, in the future, might guide the choice of treatment. PMID- 19290518 TI - Recombinantly produced hydrophobins from fungal analogues as highly surface active performance proteins. AB - Hydrophobins are available from natural resources only in milligram amounts. BASF succeeded in a recombinant production process, up-scaled to pilot plant production in kilogram scale. Strain and protein optimization by modulation of gene expression and generation of fusion proteins finally leads to two class I hydrophobins called H*Protein A and H*Protein B. By analytical ultracentrifugation, we confirm that the self-association of H*Proteins in solution is governed by their sequence, because oligomerization is induced by the same mechanisms (pH > 6, temperature >> 5 degrees C, concentration > 0.2 mg/ml) as for the well-known native hydrophobins SC3 and HFB II. Additionally, we established the triggering of structure formation by bridging with divalent ions and the stabilization of dimers and tetramers by monovalent ions or surfactants. This interplay with surfactants can be exploited synergistically: The capacity for emulsification of a 300 ppm standard surfactant solution is boosted from 0 to 100% by the addition of a mere 1 ppm of our new hydrophobins, with H*Protein A and H*Protein B having specific application profiles. This astonishing performance is rationalized by the finding that the same minute admixtures enhance significantly the interfacial elastic modulus, thus stabilizing interfaces against coalescence and phase separation. PMID- 19290519 TI - Characterization of CalS9 in the biosynthesis of UDP-xylose and the production of xylosyl-attached hybrid compound. AB - The gene cluster of calicheamicin contains calS9, which encodes UDP-GlcA decarboxylase that converts UDP-GlcA to UDP-xylose. calS9 was cloned in pET32a(+) and expressed in Escherichia coli BL21 (DE3) to characterize its putative function. The reaction product was analyzed by high-performance liquid chromatography (HPLC) and electrospray ionization-mass spectrometry. The deoxysugar biosynthesis of Streptomyces sp. KCTC 0041BP was inactivated by gene replacement to generate Streptomyces sp. GerSM2 mutant, which was unable to produce dihydrochalcomycin. calS7, calS8, and calS9 UDP-xylose biosynthetic genes were cloned in an integrative plasmid pSET152 to generate pBPDS, which was heterologously expressed in Streptomyces sp. GerSM2. Finally, novel glycosylated product, 5-O-xylosyl-chalconolide derivative, in the conjugal transformants was isolated and analyzed by HPLC and liquid chromatography-mass spectrometry. PMID- 19290520 TI - Transformation of iron sulfide to greigite by nitrite produced by oil field bacteria. AB - Nitrate, injected into oil fields, can oxidize sulfide formed by sulfate-reducing bacteria (SRB) through the action of nitrate-reducing sulfide-oxidizing bacteria (NR-SOB). When reservoir rock contains siderite (FeCO(3)), the sulfide formed is immobilized as iron sulfide minerals, e.g. mackinawite (FeS). The aim of our study was to determine the extent to which oil field NR-SOB can oxidize or transform FeS. Because no NR-SOB capable of growth with FeS were isolated, the well-characterized oil field isolate Sulfurimonas sp. strain CVO was used. When strain CVO was presented with a mixture of chemically formed FeS and dissolved sulfide (HS(-)), it only oxidized the HS(-). The FeS remained acid soluble and non-magnetic indicating that it was not transformed. In contrast, when the FeS was formed by adding FeCl(2) to a culture of SRB which gradually produced sulfide, precipitating FeS, and to which strain CVO and nitrate were subsequently added, transformation of the FeS to a magnetic, less acid-soluble form was observed. X-ray diffraction and energy-dispersive spectrometry indicated the transformed mineral to be greigite (Fe(3)S(4)). Addition of nitrite to cultures of SRB, containing microbially formed FeS, was similarly effective. Nitrite reacts chemically with HS(-) to form polysulfide and sulfur (S(0)), which then transforms SRB-formed FeS to greigite, possibly via a sulfur addition pathway (3FeS + S(0) --> Fe(3)S(4)). Further chemical transformation to pyrite (FeS(2)) is expected at higher temperatures (>60 degrees C). Hence, nitrate injection into oil fields may lead to NR-SOB-mediated and chemical mineral transformations, increasing the sulfide-binding capacity of reservoir rock. Because of mineral volume decreases, these transformations may also increase reservoir injectivity. PMID- 19290521 TI - rep-PCR fingerprinting and taxonomy based on the sequencing of the 16S rRNA gene of 54 elite commercial rhizobial strains. AB - In tropical soils, diversity and biotechnological potential of symbiotic diazotrophic bacteria are high. However, the phylogenetic relationships of prominent strains are still poorly understood. In addition, in countries such as Brazil, despite the broad use of rhizobial inoculants, molecular methods are rarely used in the analysis of strains or determination of inoculant performance. In this study, both rep-PCR (BOX) fingerprintings and the DNA sequences of the 16S rRNA gene were obtained for 54 rhizobial strains officially authorized for the production of commercial inoculants in Brazil. BOX-PCR has proven to be a reliable fingerprinting tool, reinforcing the suggestion of its applicability to track rhizobial strains in culture collections and for quality control of commercial inoculants. On the other hand, the method is not adequate for grouping or defining species or even genera. Nine strains differed in more than 1.03% (15) nucleotides of the 16S rRNA gene in relation to the closest type strain, strongly indicative of new species. Those strains were distributed across the genera Burkholderia, Rhizobium, and Bradyrhizobium. PMID- 19290522 TI - Bilateral congenital absence of the long head of the biceps tendon. AB - Absence of the long head of the biceps tendon is a rare anomaly particularly when it occurs bilaterally. We present the magnetic resonance and arthroscopy findings in a patient with bilateral congenital absence of the long head of the biceps who presented with bilateral shoulder pain. Identification of a shallow or absent intertubercular groove may aid in differentiating congenital absence of the long head of the biceps from a traumatic tendon rupture. PMID- 19290523 TI - Lipoma arborescens of the glenohumeral joint causing bone erosion: MRI features with gadolinium enhancement. AB - Lipoma arborescens is a rare benign intra-articular lesion that principally affects the knee joint. We present a case of lipoma arborescens involving the glenohumeral joint and associated with prominent large bony erosions. The gadolinium diethylenetriamine pentaacetic acid-enhanced magnetic resonance imaging (MRI) features of this lesion are also described. The characteristic MRI findings permit precise preoperative diagnosis of this rare condition even if it occurs in an atypical location and there are confusing radiological findings. PMID- 19290524 TI - A novel mutation of -50 (G-->A) in the direct repeat element of the beta-globin gene identified in a patient with severe beta-thalassemia. PMID- 19290525 TI - Repeated responses of an elderly patient with high-risk myelodysplastic syndrome to sequential therapy with tipifarnib, 5-azacitidine, and decitabine. PMID- 19290526 TI - Opposite expression pattern of Src kinase Lyn in acute and chronic haematological malignancies. AB - Lck/yes-related novel (Lyn) tyrosine kinase overexpression has been suggested to be important for leukaemic cell growth making it an attractive target for therapy. By contrast, Lyn deficiency was shown to be responsible for a phenotype resembling myeloproliferative neoplasm (MPN) in mice. We aimed to shed more light on Lyn's role in haematological neoplasm and systematically investigated Lyn expression in MPN, acute and chronic leukaemia subtypes (n = 236). On top, B-cell chronic lymphocytic leukaemia (B-CLL) and chronic myeloid leukaemia significantly overexpressed Lyn when compared to de novo acute lymphoblastic leukaemia, de novo acute myeloid leukaemia (AML) and Philadelphia-chromosome-negative myeloproliferative neoplasms (p < 0.001). Most of acute leukaemia subtypes showed a notable down-regulation of Lyn mRNA but anyhow individual cases were labelled for the active form of Lyn protein. Intriguingly, secondary AML evolved in myelodysplastic syndromes revealed almost undetectable Lyn. Overexpression of Lyn in B-CLL was associated with a significant down-regulation of microRNA-337-5p suggesting that aberrant expression of this particular microRNA could be involved in post-transcriptional control of Lyn mRNA fate. We conclude that tyrosine kinase Lyn contributes to the malignant phenotype in certain leukaemia subtypes and therefore attracts targeted therapy. PMID- 19290527 TI - Comparative analyses among the Trichomonas vaginalis, Trichomonas tenax, and Tritrichomonas foetus 5S ribosomal RNA genes. AB - The 5S ribosomal RNA (5S rRNA) is an essential component of ribosomes. Throughout evolution, variation is found among 5S rRNA genes regarding their chromosomal localization, copy number, and intergenic regions. In this report, we describe and compare the gene sequences, motifs, genomic copy number, and chromosomal localization of the Trichomonas vaginalis, Trichomonas tenax, and Tritrichomonas foetus 5S rRNA genes. T. vaginalis and T. foetus have a single type of 5S rRNA coding region, whereas two types were found in T. tenax. The sequence identities among the three organisms are between 94 and 97%. The intergenic regions are more divergent in sequence and size with characteristic species-specific motifs. The T. foetus 5S rRNA gene has larger and more complex intergenic regions, which contain either an ubiquitin gene or repeated sequences. The 5S rRNA genes were located in Trichomonads chromosomes by fluorescent in situ hybridization. PMID- 19290528 TI - Identification and characterization of differentially expressed genes from Fagus sylvatica roots after infection with Phytophthora citricola. AB - Phytophthora species are major plant pathogens infecting herbaceous and woody plants including European beech, the dominant or co-dominant tree in temperate Europe and an economically important species. For the analysis of the interaction of Phytophthora citricola with Fagus sylvatica suppression subtractive hybridization was used to isolate transcripts induced during infection and 1,149 sequences were generated. Hybridizations with driver and tester populations demonstrated differential expression in infected roots as compared to controls and verify efficient enrichment of these cDNAs during subtraction. Up regulation of selected genes during pathogenesis demonstrated using RT-PCR is consistent with these results. Pathogenesis-related proteins formed the largest group among functionally categorized transcripts. Cell wall proteins and protein kinases were also frequently found. Several transcription factors were isolated that are reactive to pathogens or wounding in other plants. The library contained a number of jasmonic acid, salicylic acid and ethylene responsive genes as well as genes directly involved in signaling pathways. Besides a mechanistic interconnection among signaling pathways another factor explaining the activation of different pathways could be the hemibiotrophic life style of Phytophthora triggering different signals in both stages. PMID- 19290529 TI - Panoramic endoluminal display with minimal image distortion using circumferential radial ray-casting for primary three-dimensional interpretation of CT colonography. AB - The purpose of this study was to develop a panoramic endoluminal display technique, the band view, which causes minimal image distortion, and to determine its feasibility as a time-efficient primary three-dimensional review method of CT colonography (CTC). Image distortion was compared between the band view and four other three-dimensional review modes using ten 10-mm and ten 20-mm electronically generated polyps. Diagnostic performance and interpretation time were compared between the band view and the conventional endoluminal view by two independent readers in 52 patients who underwent CTC and colonoscopy on the same day. Mean image distortion index values, in which 1 indicates no distortion and the larger value represents greater distortion, were significantly smaller with the band view (1.03 and 1.01 for 10-mm and 20-mm polyps, respectively) than with the filet view (1.65 and 1.55) or the virtual colon dissection (3.27 and 3.85) (P < or = 0.004). The sensitivity and specificity for detecting adenomatous polyps > or = 6 mm did not differ, but the mean interpretation time was significantly shorter with the band view than with the conventional endoluminal view by 1.8 and 4.5 minutes in readers 1 and 2, respectively (P < 0.0001). The band view can be a time-efficient alternative for primary three-dimensional review of CTC. PMID- 19290530 TI - Premotor nonspiking neurons regulate coupling among motoneurons that innervate overlapping muscle fiber population. AB - Recent work indicated that co-activity of different motoneurons (MNs) in the leech can be regulated through a network that is centered on a pair of nonspiking (NS) neurons. Here, we investigate whether this effect generalizes to different types of MNs that display differential co-activity patterns in different motor behaviors: the dorsal longitudinal excitors DE-3 and the dorsal and ventral excitors MN-L. The data indicates that both motoneurons are coupled to the NS neurons through rectifying junctions that are activated when the motoneuron membrane potential becomes more negative than that of the NS, and that they exert an inhibitory synaptic potential on NS via a polysynaptic pathway. In addition, DE-3 and MN-L are linked by junctions that allow mutual excitation but the transmission of excitatory signals from MN-L to DE-3 depended on NS membrane potential. The results support the view that NS neurons can play a central role in orchestrating the co-activity of MNs during motor behaviors. PMID- 19290531 TI - Erythromycin establishes early oral feeding in neonates operated for congenital intestinal atresias. AB - PURPOSE: The recovery of gastrointestinal function following surgery for congenital intestinal atresias can be prolonged and may increase morbidity and hospital stay. This study was conducted to investigate the prokinetic effect of erythromycin in neonates undergoing surgery for small bowel atresias. METHODS: A randomized-controlled trial was conducted at the Departments of Paediatrics and Paediatric Surgery, Military Hospital, Rawalpindi, Pakistan, from January to December 2007 to study the prokinetic effect of erythromycin (3 mg/kg per dose 4 times daily). Thirty consecutive neonates undergoing primary anastomosis for congenital small bowel atresias were randomly divided into two groups: group I (erythromycin) and group II (control). The groups were similar in terms of gestational age, sex, mode of delivery, birth weight and types of atresias. Postoperative recovery of intestinal functions was measured as time taken to achieve full enteral feed (150 ml/kg per 24 h), duration of total parenteral nutrition (TPN) and hospital stay. RESULTS: Neonates receiving oral erythromycin achieved full enteral feeding early (13.07 vs. 16.13 days) required TPN for shorter duration (10.53 vs. 13.73 days) and their hospital stay was less (16.2 vs. 18.0 days) as compared to the neonates in the control group who did not receive any erythromycin. The differences were statistically significant. CONCLUSION: The administration of oral erythromycin following primary anastomosis for small intestinal atresias results in early recovery of intestinal function, fewer days on TPN and a trend for shorter hospital stay. PMID- 19290532 TI - Long-term follow-up of a multifocal hepatic mesenchymal hamartoma producing a fetoprotein. AB - We here present a 6-month-old girl with cystic hepatic mesenchymal hamartoma and elevated a-fetoprotein (aFP). Following hepatectomy of the left lobe and partial right lobectomy, decline of the serum aFP was observed. The child has been well for 20 years and it is one among a few cases with such a long observational period. PMID- 19290533 TI - The role of prophylactic chest drainage in the operative management of esophageal atresia with tracheoesophageal fistula. AB - INTRODUCTION: Anastomotic leakage and respiratory complications are among the most common and potentially life-threatening complications following the surgical repair of esophageal atresia. Controversies exist regarding the efficacy of prophylactic extrapleural chest tube (EPCT) drainage in patients who have undergone repair of esophageal atresia. MATERIALS AND METHODS: In this prospective study, 50 newborns with esophageal atresia and distal tracheoesophageal fistula (EA-DTEF) were randomized into two groups, with no significant differences regarding gender ratio, birth body weight and delivery status. Group 1 (n = 29) underwent a right thoracotomy and a single-stage extrapleural esophageal anastomosis. Group 2 (n = 21) received additionally an intraoperative EPCT next to the anastomosis. These groups were then compared with regard to postoperative respiratory complications (such as respiratory distress, pneumonia, pneumothorax, lung collapse, and apnea), anastomotic leakage, need for mechanical ventilation, time on mechanical ventilation, and outcome. Statistical analyses were performed with Mann-Whitney U test, Fisher's exact test, and binary logistic regression analysis. RESULTS: The rates of respiratory complications and anastomotic leakage, need for mechanical ventilation, time on mechanical ventilation, and mortality rate were comparable between the two groups (P > 0.05). Mortality was associated with respiratory complications (P = 0.003) and anastomotic leakage (P = 0.007). CONCLUSION: It seems that prophylactic EPCT drainage does not decrease the early postoperative respiratory complications and mortality rates in newborns with EA-DTEF. PMID- 19290534 TI - Diffuse intraneural leiomyoma in a case of sensorimotor neuropathy. PMID- 19290535 TI - Photodynamic therapy with meta-tetrahydroxyphenylchlorin (Foscan) in the management of squamous cell carcinoma of the head and neck: experience with 35 patients. AB - Photodynamic therapy (PDT) is a relatively new method of treating superficial tumours of the skin and mucosa. After the injection of a photosensitizing agent, the tumour area is exposed to non-thermal laser light. This causes a phototoxic reaction, producing oxygen radicals that destroy tumour cells. From November 2003 to July 2007, a total of 35 patients with recurrent squamous cell carcinoma or secondary tumours of the head and neck region were treated with PDT at the German Armed Forces Hospital in Ulm. These patients had failed or found unsuitable for other treatments. Meta-tetrahydroxyphenylchlorin (mTHPC), known under the trade name of Foscan, was used as the photosensitizing agent. Local control was achieved in 21 patients (60%) and partial remission in 10 patients (28.5%). Four patients (11.5%) did not respond to PDT treatment. The mean duration of overall survival was 401.45 (+/-321.2) days, median was 356 after the completion of treatment. The mean duration of recurrence-free survival was 327.7 (+/-131.1) days, median was 181 for patients with complete remission. None of the patient developed serious complications. Photodynamic therapy is an important treatment option for patients who present with recurrent carcinoma or secondary tumours of the upper aerodigestive tract and who have failed or unsuitable for other treatments. Due to the excellent treatment results that have been achieved so far, PDT may in the future also play a role in the primary treatment of superficial tumours of the oral cavity, pharynx and larynx. PMID- 19290536 TI - Relative biological effectiveness of 25 and 10 kV X-rays for the induction of chromosomal aberrations in two human mammary epithelial cell lines. AB - Administration of ionizing radiation for diagnostic purposes can be associated with a risk for the induction of tumors. Therefore, particularly with regard to general screening programs, e.g. with mammography, cost-benefit considerations must be discussed including risk estimation depending upon the radiation quality administered. The present study was initiated to investigate the in vitro X-ray energy dependence for the induction of chromosomal aberrations in the two mammary epithelial cell lines, 184A1 and MCF-12A. The induced excess fragments, dicentric chromosomes and centric rings were analyzed and the relative biological effectiveness (RBE) was determined for 10 and 25 kV X-rays relative to 200 kV X rays. The assumed energy dependence with higher values for 10 kV X-rays was confirmed for the excess fragments, with RBE(M) values of 1.92 +/- 0.26 and 1.40 +/- 0.12 for 10 kV X-rays and 1.17 +/- 0.12 and 0.97 +/- 0.10 for 25 kV photons determined for cell lines 184A1 and MCF-12A, respectively. Meaningful results for the induction of dicentric chromosomes and centric rings were obtained only for higher doses with RBE values of 1.31 +/- 0.21 and 1.70 +/- 0.29 for 184A1 and 1.08 +/- 0.08 and 1.43 +/- 0.12 for MCF-12A irradiated with 25 and 10 kV X-rays, respectively. PMID- 19290537 TI - Effect of epigallocatechin-gallate on inner retinal function in ocular hypertension and glaucoma: a short-term study by pattern electroretinogram. AB - BACKGROUND: Epigallocatechin-gallate (EGCG) is a powerful antioxidant with suggested neuroprotective action. The aim of this study was to evaluate the effect of short-term supplementation of EGCG on inner retinal function in ocular hypertension (OHT) and open-angle glaucoma (OAG). METHODS: Eighteen OHT and 18 OAG patients (perimetric mean deviation: >-10 dB) were randomly assigned to assume oral placebo or EGCG over a 3-month period in a randomized, placebo controlled, double-blind, cross-over design clinical trial (clinicaltrials.gov identifier: NCT00476138). Pattern-evoked electroretinograms (PERGs) to 1.6 cycles/degree square-wave gratings, counterphased at 16 reversals/second, and standard automated perimetry (Humphrey 30-2) were assessed at the study entry (baseline), and after 3 months of placebo or EGCG. RESULTS: After EGCG, PERGs of OAG, but not OHT patients were increased in amplitude, compared either to baseline values (mean amplitude change: 0.06 log microV, p < 0.05) or to PERG amplitude values found in the same patients after placebo administration (mean change: -0.02 log microV, p not significant; difference between EGCG and placebo: 0.08 log microV, p < 0.05). In both OHT and OAG patients, standard automated perimetry did not show significant changes after either EGCG or placebo. In individual OAG patients, the magnitude of PERG amplitude increment after EGCG was inversely related (r = -0.8, p < 0.01) to corresponding baseline amplitudes. CONCLUSIONS: Although this study cannot provide evidence for long-term benefit of EGCG supplementation in OAG, and the observed effect is small, the results suggest that EGCG might favourably influence inner retinal function in eyes with early to moderately advanced glaucomatous damage. PMID- 19290538 TI - Spatiotemporal distribution patterns of oligosaccharides during early embryogenesis in the starfish Patiria pectinifera. AB - To examine embryogenic mechanisms in the starfish Patiria (Asterina) pectinifera, we histochemically analyzed several larval stages using Alcian Blue (AB, which stains acidic mucins), Periodic Acid Schiff (PAS, which stains neutral mucins), and 21 types of lectins. Carbohydrate distribution patterns were observed in the cytoplasm, basement membrane, and blastocoel as follows: (1) The first group of lectins showed granular signals in the mesendodermal cells, and these lectins may be useful as mesendoderm markers. (2) The second class of lectins showed diffuse signals across the entire cytoplasm from the hatched blastula until the mid gastrula. These signals became localized to the basal cytoplasm of archenteron cells at the early bipinnaria. (3) Lectin reactivity in the basement membrane peaked at the early-to-mid gastrula and was nearly gone by the early bipinnaria. These results suggest the existence of various substances in the basement membrane and imply the importance of these substances during archenteron elongation and the induction of mesenchyme differentiation. (4) Signal colors with AB-PAS double staining in the blastocoel changed from magenta (by PAS staining) into blue (by AB staining) during these stages, thus, indicating that mucin located in the blastocoel changed from neutral to acidic. The most significant part of this report is the first description regarding temporal changes in the characteristics of intra- and extracellular components with the combination of many different lectins and stains. PMID- 19290539 TI - Genetic identification of Fasciola hepatica by ITS-2 sequence of nuclear ribosomal DNA in Turkey. AB - The trematodes of the genus Fasciola are the common liver flukes of a range species of animals and have a global geographical distribution. They can generally be distinguished on the basic of their morphology. ITS-2 ribosomal DNA sequences have been used to characterize the liver flukes as a specific marker from different geographical regions which include F. hepatica, F. gigantica, and an intermediate Fasciola. To determine the phylogenetic location of F. hepatica of Turkey origin based on ITS-2 rDNA molecular data, adult F. hepatica trematodes were collected from the liver naturally infected sheep from different geographical locations in Turkey (Elazig, Malatya, Samsun). ITS-2 rDNA were cloned, sequenced, and compared with published sequences ITS-2 rDNA of other species of trematodes in the family Fasciolidae using the GenBank Blast program. The only one ITS-2 sequence had defined for the examined Turkish F. hepatica samples. The phylogenetic trees constructed based upon the ITS-2 sequences from Turkey by multiple tree-building methods in MEGA revealed a close relationship with isolates of F. hepatica, F. gigantica, and Fasciola sp. The present study is the first demonstration of the existence of F. hepatica in sheep in Turkey by the genetic approach using ITS-2 rDNA as genetic marker. PMID- 19290540 TI - Molecular epidemiology of human Blastocystis isolates in France. AB - Blastocystis sp. is the most common eukaryotic parasite in the intestinal tract of humans. Due to its strong impact in public health, in this study, we determined the frequency of different Blastocystis subtypes in patients in France. We hypothesized on the mode of transmission and tested a possible relationship between the subtype and symptomatic status. We obtained a total of 40 stool samples identified as positive for Blastocystis by microscopic examination of smears. Participants consisted of 25 symptomatic and 15 asymptomatic patients, for whom clinical and parasitological data were collected. For nested-polymerase chain reaction and genotyping, DNA was extracted directly from fecal samples or from fecal cultures. Morphological forms observed in fecal cultures were uncorrelated with symptomatic status. Genotyping using partial small subunit rRNA gene analysis identified a total of 43 Blastocystis isolates corresponding to 37 single infections and three mixed infections by two different subtypes. These 43 isolates belonged to five subtypes (1, 2, 3, 4, and 7) with predominance of subtype 3 (53.5%). Patient symptomatic status was uncorrelated with Blastocystis subtype. PMID- 19290541 TI - Schistosoma japonicum: proteomics analysis of differentially expressed proteins from ultraviolet-attenuated cercariae compared to normal cercariae. AB - Schistosomiasis is considered the most important human helminthiasis in terms of morbidity and mortality. In this study, comparative soluble proteomic analysis of normal cercariae and ultraviolet-irradiated attenuated cercariae (UVAC) from Schistosoma japonicum were carried out in view of the high efficiency of irradiation-attenuated cercariae vaccine. The results revealed that some proteins showed significant differential expression in the parasite after treatment with ultraviolet light. Total 20 protein spots were identified by mass spectrometry, corresponded to five groups according to their functions in the main that were structural and motor proteins (actin, et al.), energy metabolism associated enzymes (glyceraldehydes-3-phosphage dehydrogenase, et al.), signaling transduction pathway-associated molecules (14-3-3 protein, et al.), heat shock protein families (HSP 70 family, et al.), and other functional proteins (20S proteasome). Furthermore, our results indicated that the differential expression of the proteins by ultraviolet irradiation may be, at least partially, acquired by regulating the mRNA levels of corresponding proteins. These results may provide new clues for further exploring the mechanism of protective immunity induced by UVAC and may shed some light on the development of vaccines against schistosomiasis. PMID- 19290543 TI - Insensitivity of chloroplast gene expression to DNA methylation. AB - Presence and possible functions of DNA methylation in plastid genomes of higher plants have been highly controversial. While a number of studies presented evidence for the occurrence of both cytosine and adenine methylation in plastid genomes and proposed a role of cytosine methylation in the transcriptional regulation of plastid genes, several recent studies suggested that at least cytosine methylation may be absent from higher plant plastid genomes. To test if either adenine or cytosine methylation can play a regulatory role in plastid gene expression, we have introduced cyanobacterial genes for adenine and cytosine DNA methyltransferases (methylases) into the tobacco plastid genome by chloroplast transformation. Using DNA cleavage with methylation-sensitive and methylation dependent restriction endonucleases, we show that the plastid genomes in the transplastomic plants are efficiently methylated. All transplastomic lines are phenotypically indistinguishable from wild-type plants and, moreover, show no alterations in plastid gene expression. Our data indicate that the expression of plastid genes is not sensitive to DNA methylation and, hence, suggest that DNA methylation is unlikely to be involved in the transcriptional regulation of plastid gene expression. PMID- 19290544 TI - African ancestry is associated with risk of asthma and high total serum IgE in a population from the Caribbean Coast of Colombia. AB - African descended populations exhibit an increased prevalence of asthma and allergies compared to Europeans. One approach to distinguish between environmental and genetic explanations for this difference is to study relationships of asthma risk to individual admixture. We aimed to determine the admixture proportions of a case-control sample from the Caribbean Coast of Colombia currently participating in genetic studies for asthma, and to test for population stratification and association between African ancestry and asthma and total serum IgE levels (tIgE). We genotyped 368 asthmatics and 365 non-asthmatics for 52 autosomal ancestry informative markers, six mtDNA haplogroups and nine haplogroups and five microsatellites in Y chromosome. Autosomal admixture proportions, population stratification, and associations between ancestry and the phenotypes were estimated by ADMIXMAP. The average admixture proportions among asthmatics were 42.8% European, 39.9% African and 17.2% Native American and among non-asthmatics they were 44.2% (P = 0.068), 37.6% (P = 0.007) and 18.1% (P = 0.050), respectively. In the total sample, the paternal contributions were 71% European, 25% African and 4.0% Native American and the maternal lineages were 56.8% Native American, and 20.2% African; 22.9% of the individuals carried other non-Native American mtDNA haplogroups. African ancestry was significantly associated with asthma (OR: 2.97; 95% CI: 1.08-8.08), high tIgE (OR: 1.9; 95% CI: 1.17-3.12) and socioeconomic status (OR = 0.64; 95% CI: 0.47-0.87). Significant population stratification was observed in this sample. Our findings indicate that genetic factors can explain the association between asthma and African ancestry and suggest that this sample is a useful resource for performing admixture mapping for asthma. PMID- 19290545 TI - Glycine regulates proliferation and differentiation of salivary-gland-derived progenitor cells. AB - Amino acids have various physiological activities that influence processes such as intestinal regeneration, EGF secretion, protein synthesis, and cell growth. Salivary glands are exposed to nutrients that influence their proliferation and regeneration. Glycine is included in saliva in large quantities and reportedly has important roles in antibacterial activities and the inhibition of tumor growth and as a precursor of nucleotide synthesis in cell proliferation. We have investigated the effects of glycine on the proliferation and differentiation of salivary glands by using mouse salivary-gland-derived progenitor (mSGP) cells. In cultures of mSGP cells, cell proliferation is suppressed in the presence of glycine, whereas it is promoted by its removal. Glycine promotes three dimensional formations of mSGP cells, which are negative for immature markers and positive for differentiation markers. In cell-cycle analysis, cell-cycle progression is delayed at the S-phase by glycine supplementation. Glycine also suppresses the phosphorylation of p42/p44MAPK. These results suggest that glycine suppresses the proliferation and promotes the differentiation of mSGP cells, and that it has inhibitory effects on growth factor signaling and cell-cycle progression. Glycine might therefore be a physiological activator that regulates the proliferation and differentiation of salivary glands. PMID- 19290546 TI - Hoxa3 and signaling molecules involved in aortic arch patterning and remodeling. AB - Anomalies of the aortic arch have long been of anatomicoclinical interest. Recent studies on gene-targeted mice have identified the candidate genes that are involved in the patterning and remodeling of the pharyngeal arch arteries. In this review, we discuss our present knowledge with regard to the signaling molecules that regulate specific aspects of arch artery development. We focus first on Hoxa3, because it plays a critical role in the regulation of the differentiation of the third pharyngeal arch. Hoxa3 is expressed by the neural crest cells that originate from the rhombomeres, viz., (r)5, r6, and r7, and populate the third pharyngeal arch; it is also expressed in the third pharyngeal pouch. In Hoxa3 homozygous null mutant mice, the third arch artery degenerates bilaterally at embryonic day 11.5, resulting in the malformation of the carotid artery system. Complex combinatorial signals among the neural crest cells, pharyngeal mesoderm, ectoderm, and pouch endoderm are required for the proper development of the arch arterial system. Therefore, we highlight the numerous signaling pathways and individual genes expressed by the ectomesenchymal neural crest cells and also by the other epithelial and mesodermal cells of the pharynx. Defects in these genes result in malformations of the arch artery derivatives. This review should deepen our understanding of congenital human syndromes with abnormal patterns of pharyngeal arch arteries. PMID- 19290547 TI - Influence of hydrologic attributes on brown trout recruitment in low-latitude range margins. AB - Factors controlling brown trout Salmo trutta recruitment in Mediterranean areas are largely unknown, despite the relevance this may have for fisheries management. The effect of hydrological variability on survival of young brown trout was studied during seven consecutive years in five resident populations from the southern range of the species distribution. Recruit density at the end of summer varied markedly among year-classes and rivers during the study period. Previous work showed that egg density the previous fall did not account for more than 50% of the observed variation in recruitment density. Thus, we expected that climatic patterns, as determinants of discharge and water temperature, would play a role in the control of young trout abundance. We tested this by analyzing the effects of flow variation and predictability on young trout survival during the spawning to emergence and the summer drought periods. Both hatching and emergence times and length of hatching and emergence periods were similar between years within each river but varied considerably among populations, due to differences in water temperature. Interannual variation in flow attributes during spawning to emergence and summer drought affected juvenile survival in all populations, once the effect of endogenous factors was removed. Survival rate was significantly related to the timing, magnitude and duration of extreme water conditions, and to the rate of change in discharge during hatching and emergence times in most rivers. The magnitude and duration of low flows during summer drought appeared to be a critical factor for survival of young trout. Our findings suggest that density-independent factors, i.e., hydrological variability, play a central role in the population dynamics of brown trout in populations from low-latitude range margins. Reported effects of hydrologic attributes on trout survival are likely to be increasingly important if, as predicted, climate change leads to greater extremes and variability of flow regimes. PMID- 19290548 TI - Improve the management of patients with skull bone metastases by means of helical tomotherapy. PMID- 19290549 TI - Reducing gastric secretions--a role for histamine 2 antagonists or proton pump inhibitors in malignant bowel obstruction? AB - GOALS OF WORK: Inoperable bowel obstructions are not uncommon in advanced cancer and are associated with a very poor prognosis. Symptom control includes reducing the frequency of vomiting by prescription of antisecretory medications. The most commonly used agents for this are either hyoscine butylbromide or octreotide. Either histamine 2 antagonists or proton pump inhibitors are sometimes recommended as adjuvants to reduce gastric secretions. The aim of this study was to examine the effects of histamine 2 antagonists and proton pump inhibitors and to objectively compare the effects of one agent over another. MATERIALS AND METHODS: Previously, electronic databases were searched for trials that compared ranitidine versus proton pump inhibitors in their effect on volume of gastric aspirates. RESULTS: Seven trials were included in a meta-analysis. Pooled outcomes suggest that both proton pump inhibitors and ranitidine reduce gastric volumes, but the most superior agent is ranitidine, which reduces the volume of gastric secretions by an average of 0.22 ml.kg(-1); 95% confidence interval 0.04 to 0.41. CONCLUSIONS: Based on well-conducted studies, objective evidence exists that confirms ranitidine will decrease the volume of gastric aspirates. This forms a sound basis from which to develop further research aimed at improving the care of people with malignant bowel obstructions. PMID- 19290550 TI - Cancer-associated hypercalcemia: validation of a bedside prognostic score. AB - INTRODUCTION: Until now, there was no validated and reliable tool to estimate life expectancy among patients with cancer-associated hypercalcemia (CAH). This study aimed to validate a prognostic score previously published. METHODS: We studied a new cohort of 252 patients with CAH. Our prognostic score based on the four poor-prognosis factors (liver metastasis, squamous cell carcinoma, hypoalbuminemia, and calcemia >2.83 mmol) was implemented to this cohort. The accuracy of this score for the prediction of 90-day mortality was estimated with area under the curve of the receiver operator curve (AUC-ROC). RESULTS: Among this validation cohort, patients with score = 0 experienced a median overall survival of 797 days; whereas, patients with score >/=1 (at least one poor prognosis factor) had median overall survival of 49 days (p < 0.0001). The AUC ROC for the prediction of 90-day mortality were 0.88 (0.76-0.86) and 0.67 (0.61 0.74) in the development cohort and in the validation cohort, respectively. CONCLUSION: We have validated a reliable bedside prognostic score for CAH. PMID- 19290551 TI - Ternary oxovanadium(IV) complexes of ONO-donor Schiff base and polypyridyl derivatives as protein tyrosine phosphatase inhibitors: synthesis, characterization, and biological activities. AB - A series of oxovanadium complexes with mixed ligands, a tridentate ONO-donor Schiff base ligand [viz., salicylidene anthranilic acid (SAA)], and a bidentate NN ligand [viz., 2,2'-bipyridine (bpy), 1,10-phenanthroline (phen), dipyrido[3,2 d:2',3'-f]quinoxaline (dpq), dipyrido[3,2-a:2',3'-c]phenazine (dppz), or 7 methyldipyrido[3,2-a:2',3'-c]phenazine (dppm)], have been synthesized and characterized by elemental analysis, electrospray ionization mass spectrometry, UV-vis spectroscopy, Fourier transform IR spectroscopy, EPR spectroscopy, and X ray crystallography. Crystal structures of both complexes, [V(IV)O(SAA)(bpy)].0.25bpy and [V(IV)O(SAA)(phen)].0.33H(2)O, reveal that oxovanadium(IV) is coordinated with one nitrogen and two oxygen atoms from the Schiff base and two nitrogen atoms from the bidentate planar ligands, in a distorted octahedral geometry (VO(3)N(3)). The oxidation state of V(IV) with d(1) configuration was confirmed by EPR spectroscopy. The speciation of VO-SAA-bpy in aqueous solution was investigated by potentiomtreic pH titrations, and the results revealed that the main species are two ternary complexes at a pH range of 7.0-7.4, and one is the isolated crystalline complex. The complexes have been found to be potent inhibitors against human protein tyrosine phosphatase 1B (PTP1B) (IC(50) approximately 30-61 nM), T-cell protein tyrosine phosphatase (TCPTP), and Src homology phosphatase 1 (SHP-1) in vitro. Interestingly, the [V(IV)O(SAA)(bpy)] complex selectively inhibits PTP1B over the other two phosphatases (approximate ninefold selectivity against SHP-1 and about twofold selectivity against TCPTP). Kinetics assays suggest that the complexes inhibit PTP1B in a competitive and reversible manner. These suggest that the complexes may be promising candidates as novel antidiabetic agents. PMID- 19290552 TI - Mechanistic insight into the initiation step of the reaction of Burkholderia pseudomallei catalase-peroxidase with peroxyacetic acid. AB - The reaction of the catalase-peroxidase of Burkholderia pseudomallei with peroxyacetic acid has been analyzed using stopped-flow spectrophotometry. Two well-defined species were observed, the first defined by an increase in intensity and narrowing of the Soret band at 407 nm and a 10-nm shift of the charge transfer band from 635 to 625 nm. These features are consistent with a ferric spectrum with a greater proportion of sixth-coordination character and are assigned to an Fe(III)-peroxyacetic acid complex. Complementary 9-GHz EPR characterization of the changes in the ferric signal of the resting enzyme induced by the binding of acetate in the heme pocket substantiates the proposal. Kinetic analysis of the spectral changes as a function of peroxyacetic acid concentration revealed two independent peroxyacetic acid binding events, one coincident with formation of the Fe(III)-peroxyacetic acid complex and the other coincident with the heme oxidation to the subsequent ferryl intermediate. A model to explain the need for two peroxyacetic acid binding events is proposed. The reaction of the W330F variant followed similar kinetics, although the characteristic spectral features of the Fe(IV)=O Por(*+) species were detected. The variant D141A lacking an aspartate at the entrance to the heme cavity as well as the R108A and D141A/R108A variants showed no evidence for the Fe(III) peroxyacetic acid complex, only the formation of ferryl species with absorbance maxima at 414, 545, and 585 nm. PMID- 19290553 TI - Insight into the protein and solvent contributions to the reduction potentials of [4Fe-4S]2+/+ clusters: crystal structures of the Allochromatium vinosum ferredoxin variants C57A and V13G and the homologous Escherichia coli ferredoxin. AB - The crystal structures of the C57A and V13G molecular variants of Allochromatium vinosum 2[4Fe-4S] ferredoxin (AlvinFd) and that of the homologous ferredoxin from Escherichia coli (EcFd) have been determined at 1.05-, 1.48-, and 1.65-A resolution, respectively. The present structures combined with cyclic voltammetry studies establish clear effects of the degree of exposure of the cluster with the lowest reduction potential (cluster I) towards less negative reduction potentials (E degrees ). This is better illustrated by V13G AlvinFd (high exposure, E degrees = -594 mV) and EcFd (low exposure, E degrees = -675 mV). In C57A AlvinFd, the movement of the protein backbone, as a result of replacing the noncoordinating Cys57 by Ala, leads to a +50-mV upshift of the potential of the nearby cluster I, by removal of polar interactions involving the thiolate group and adjustment of the hydrogen-bond network involving the cluster atoms. In addition, the present structures and other previously reported accurate structures of this family of ferredoxins indicate that polar interactions of side chains and water molecules with cluster II sulfur atoms, which are absent in the environment of cluster I, are correlated to the approximately 180-250 mV difference between the reduction potentials of clusters I and II. These findings provide insight into the significant effects of subtle structural differences of the protein and solvent environment around the clusters of [4Fe-4S] ferredoxins on their electrochemical properties. PMID- 19290554 TI - Human serum transferrin: a tale of two lobes. Urea gel and steady state fluorescence analysis of recombinant transferrins as a function of pH, time, and the soluble portion of the transferrin receptor. AB - Iron release from human serum transferrin (hTF) has been studied extensively; however, the molecular details of the mechanism(s) remain incomplete. This is in part due to the complexity of this process, which is influenced by lobe-lobe interactions, the transferrin receptor (TFR), the salt effect, the presence of a chelator, and acidification within the endosome, resulting in iron release. The present work brings together many of the concepts and assertions derived from previous studies in a methodical, uniform, and visual manner. Examination of earlier work reveals some uncertainty due to sample and technical limitations. We have used a combination of steady-state fluorescence and urea gels to evaluate the effect of conformation, pH, time, and the soluble portion of the TFR (sTFR) on iron release from each lobe of hTF. The use of authentic recombinant monoferric and locked species removes any possibility of cross-contamination by acquisition of iron. Elimination of detergent by use of the sTFR provides a further technical advantage. We find that iron release from the N-lobe is very sensitive to the conformation of the C-lobe, but is insensitive to the presence of the sTFR or to changes in pH (between 5.6 and 6.4). Specifically, when the cleft of the C-lobe is locked, the urea gels indicate that only about half of the iron is completely removed from the cleft of the N-lobe. Iron release from the C lobe is most affected by the presence of the sTFR and changes in pH, but is unaffected by the conformation of the N-lobe. A model for iron release from diferric hTF is provided to delineate our findings. PMID- 19290555 TI - Computational modeling of the adsorption and (*)OH initiated photochemical and photocatalytic primary oxidation of nitrobenzene. AB - The adsorption and primary oxidation step for the photodegradation of nitrobenzene (NB) have been studied computationally using MSINDO SCF MO method. The method performs efficiently for extended surface models such as Ti(36)O(90)H(36). Molecular dynamics simulations have revealed that NB is linked to TiO(2) surface at the titanium ion via the oxygen atoms of NO(2) group. In addition, the computed vibrational density of states for the adsorbed NB molecule is in reasonably good agreement with the available experimental data and theoretical results. In order to identify the primary photochemical and photocatalytic (*)OH initiated photooxidation intermediates, we have employed two different theoretical approaches, frontier orbital theory and Wheland localization theory. It has been found that the meta- hydroxynitrocyclohexadienyl radical is energetically more favored than para- and ortho hydroxynitrocyclohexadienyl radicals for the photochemical photolysis, whereas in the case of photocatalysis, the (*)OH radical attack is unselective and all three possible isomers have comparable stabilities. PMID- 19290557 TI - Procedures and methods of benefit assessments for medicines in Germany: give the child a name. PMID- 19290556 TI - Identification of the variant Ala335Val of MED25 as responsible for CMT2B2: molecular data, functional studies of the SH3 recognition motif and correlation between wild-type MED25 and PMP22 RNA levels in CMT1A animal models. AB - Charcot-Marie-Tooth (CMT) disease is a clinically and genetically heterogeneous disorder. All mendelian patterns of inheritance have been described. We identified a homozygous p.A335V mutation in the MED25 gene in an extended Costa Rican family with autosomal recessively inherited Charcot-Marie-Tooth neuropathy linked to the CMT2B2 locus in chromosome 19q13.3. MED25, also known as ARC92 and ACID1, is a subunit of the human activator-recruited cofactor (ARC), a family of large transcriptional coactivator complexes related to the yeast Mediator. MED25 was identified by virtue of functional association with the activator domains of multiple cellular and viral transcriptional activators. Its exact physiological function in transcriptional regulation remains obscure. The CMT2B2-associated missense amino acid substitution p.A335V is located in a proline-rich region with high affinity for SH3 domains of the Abelson type. The mutation causes a decrease in binding specificity leading to the recognition of a broader range of SH3 domain proteins. Furthermore, Med25 is coordinately expressed with Pmp22 gene dosage and expression in transgenic mice and rats. These results suggest a potential role of this protein in the molecular etiology of CMT2B2 and suggest a potential, more general role of MED25 in gene dosage sensitive peripheral neuropathy pathogenesis. PMID- 19290558 TI - Inflammation status of rabbit carotid artery model endothelium during intermittent hypoxia exposure and its relationship with leptin. AB - OBJECTIVES: To develop an intermittent hypoxia/reoxygenation (IH/ROX) rabbit carotid artery model and then investigate the inflammation status of rabbit carotid artery endothelium after IH exposure and its relationship with leptin. MATERIALS AND METHODS: After anesthetization, rabbit's right common carotid artery was cleared of surrounding tissue with anatomic microscope, cannulated to its distal part and the proximal part was ligated. Preparations were challenged by changing the PO(2) of the gas mixture equilibrating the perfusate. Alternate perfusing (2 mL/min) of equilibrated perfusate bubbled with normoxia or hypoxia gas mixtures formed IH/ROX cycles in the right carotid common artery, simulating the pattern of hypoxic episodes seen in obstructive sleep apnea (OSA), or continuous perfusing of hypoxia perfusate to form continuous hypoxia (CH) modes. Sixty adult male New Zealand White rabbits (2.5-3.0 kg) were separated into six groups, ten per group. Groups were: A, intermittent normoxia (IN) group, perfused with perfusion equilibrated with 21% O(2) [PO(2) about 141 +/- 2.87 mmHg] for 15 s and 21% O(2) for 1 min 45 s, 60 cycles; B, severe IH group, 5% O(2) [PO(2) about 35.2 +/- 1.27 mmHg] 15 s and 21% O(2) 1 min 45 s, 60 cycles; C, mild IH group, 10% O(2) [PO(2) about 54.3 +/- 3.31 mmHg] 15 s and 21% O(2) 1 min 45 s, 60 cycles; D, severe IH+Lep group, protocol was the same with severe IH group; E, CH group, IN for 1 h 45 min and then 5% O(2) for 15 min; and F, Lep group, the same with IN group. Right common carotid artery parts distal to the cannula were harvested after exposure, and endothelial cell layers were gotten from longitudinal outspread vessels. Nuclear factor kappaB (NFkappaB) DNA binding activities of partial cell layers were measured with electrophoretic mobility shift assay in the IN group, severe IH group, mild IH group, and CH group nuclear extracts. The other part of the cell layers in the IN group, severe IH group, severe IH+Lep group, and Lep group were cultured for 2 h, and during the culture procedure, recombinated human leptin solutions were added to culture dishes of severe IH+Lep group and Lep group (resulted concentration, 10 ng/mL). Enzyme linked immunosorbent assay was used to analyze medium interleukin-6 (IL-6) concentrations, reverse transcription polymerase chain reaction was used to analyze endothelial cell Ras homology A (RhoA) mRNA expression levels. Statistical analysis was done with SPSS 11.5 software package. RESULTS: NFkappaB DNA binding activities were significantly different between groups (F = 112.428, P < 0.001). This activity in the severe IH group (4.27 +/- 0.64) was higher than that in the mild IH group (2.33 +/- 0.45, P < 0.001), IN group (1.00 +/- 0.26, P < 0.001), and CH group (1.15 +/- 0.36, P < 0.001). RhoA mRNA expression levels were different in groups (F = 26.634, P < 0.001).This level in the severe IH+Lep group (2.54 +/- 0.53) was higher than that in the severe IH group (1.57 +/- 0.44, P = 0.002), IN group (1.00 +/- 0.31, P < 0.001), and Lep group (1.31 +/- 0.30, P < 0.001). IL-6 concentrations were different in groups (F = 79.922, P < 0.001). IL-6 concentration in the severe IH+Lep group (1591.50 +/- 179.57 pg/mL) was higher than that in the severe IH group (1217.20 +/- 320.62 pg/mL, P = 0.036), IN group (325.40 +/- 85.26 pg/mL, P < 0.001), and Lep group (517.40 +/- 183.09 pg/mL, P < 0.001). CONCLUSIONS: IH/ROX activated the inflammation pathway significantly in the endothelium, which was more intensive than CH and intensity dependent. When exposed to both IH/ROX and leptin, inflammation occurs more dramatically. It means that synergic activating roles were performed by IH/ROX and leptin. This study may have a clinical implication that IH can cause endothelial damage through activated inflammation in OSA patients, and if the OSA patients have obesity at the same time, the endothelial damage or the inflammation would be more significant because of elevated leptin level as a synergic factor. PMID- 19290559 TI - Seasonal differences in mercury accumulation in Trichiurus lepturus (Cutlassfish) in relation to length and weight in a Northeast Brazilian estuary. AB - BACKGROUND, AIM, AND SCOPE: At tropical latitudes, and especially on the semi arid coasts of the Brazilian Northeast, the rainfall regime governs the water quality of estuaries due to the pronounced difference between the rainy and dry seasons. These changes may be responsible for seasonal changes in bioavailability of mercury (Hg) and other pollutants to the estuarine and coastal biota. Mercury bioaccumulates along estuarine-marine food chains usually result in higher concentrations in tissues of top predators and posing a risk to both marine mammals and humans alike. The Goiana River Estuary (7.5 degrees S) is a typical estuary of the semi-arid tropical regions and supports traditional communities with fisheries (mollusks, fish, and crustacean). It is also responsible for an important part of the biological production of the adjacent coastal waters. MATERIALS AND METHODS: Trichiurus lepturus (Actinopterygii: Perciformes) is a pscivorous marine straggler. Fish from this species (n = 104) were captured in a trapping barrier used by the local traditional population and using an otter trawl net along the main channel of the low estuary during two dry seasons (D1 = November, December 2005, January 2006; D2 = November, December 2006, January 2007) and the end of a rainy season (R = August, September, October 2006). Fish muscle samples were preserved cold and then freeze-dried prior to analysis of its total mercury (Hg-T) contents. Total mercury was determined by cold vapor atomic absorption spectrometry (CV-AAS) with sodium borohydride as a reducing agent. RESULTS: The studied individuals (n = 104) were sub-adult (30-70 cm, 71 ind.) and adult fish (>70 cm, 33 ind.). Weight (W) (204.1 +/- 97.9 g, total biomass = 21,229.7 g) and total length (TL) (63.1 +/- 10.1 cm, range 29.5-89.0 cm) presented a significant (p < 0.05) correlation. Two-way ANOVA (n = 81) showed that TL and W had significant differences (p < 0.05) among seasons, being higher in D1 than in D2 and R, respectively. Moreover, season vs. month interaction were detected for the variables length and weight. For the variable weight was detected significant difference for the factor month (p < 0.05). It suggests that the fish enter the estuary at the end of the rainy season and increase in length and weight during the time they spend in the estuary. Fish from this estuary are shown to be fit for human consumption (125.3 +/- 61.9 microgHg-T kg(-1) w.wt.; n = 104). Fish mercury contents increased with size and weight. Correlations between TL and Hg-T (r = 0.37286) and between W and Hg-T (r = 0.38212) were significant (p < 0.05). Dryer months showed higher mercury concentrations in fish (D1 773.4 +/- 207.5 microgHg-T kg(-1) d.wt., n = 27; D2 370.1 +/- 78.8 microgHg-T kg(-1) d.wt., n = 27; R 331.2 +/- 138.5 microgHg-T kg(-1) d.wt., n = 27). The variable mercury concentration showed differences in relation to the factor season (p < 0.05), where fish captured during the first dry season showed the highest concentration of mercury. The correlation between Hg-T and rainfall (Rf) showed a negative correlation (r = -0.56; p < 0.05). DISCUSSION: The main likely source of mercury to this estuary is diffuse continental run off, including urban and industrial effluents. Since concentration of mercury in fish tissue is negatively correlated to rainfall, but positively correlated with fish length and weight, it suggests that fish growth in this estuary results in mercury uptake and concentration on the fish tissue. In the dry season of 2005-2006, when rainfall remained below the historic average, fish bioaccumulated significantly more mercury than in the dry season 2006-2007, when rainfall was within the predictable historic average. It is suggested that less rainfall, and consequently less particulate matter and less primary production in the estuary, make mercury more available to the higher levels of the estuarine food chain. In the case of higher rainfall, when river flow increases and water quality in the estuary is reduced, mercury probably is quickly exported associated to the particulate matter to the adjacent coastal waters where it then disperses. This species is a potential routine bioindicator for mercury contamination of the biota, but so far was used only with a limited number of individuals and contexts. CONCLUSIONS: Fish from the Goiana River estuary can still be safely consumed by the local population. However, any further contamination of this resource might lead to total mercury levels above the recommended limits for pregnant women and small children. The proposed heavy dependency of total mercury levels in fish on water quality indicates that land use and water quality standards must be more closely watched in order to guarantee that best possible practices are in place to prevent bioaccumulation of mercury and its transfer along the food chain. Human interventions and climatic events which affect river water flow are also playing a role in the mercury cycle at tropical semi-arid estuaries. RECOMMENDATIONS AND PERSPECTIVES: T. lepturus is largely consumed by coastal populations of tropical and sub-tropical countries all over the world. It is also consumed by a number of marine mammals over which we have a strong conservation interest. This species is also a link among different ecosystems along the estuarine ecocline. Therefore, knowledge of its degree of contamination might contribute to public health issues as well as marine conservation actions. Studies on mercury and other contaminants using this species as bioindicator (cosmopolitan, readily available) could help elucidating mechanisms through which pollutants are being transferred not only through the food chain, but also from estuarine-coastal-open waters. In addition, using the same species in marine pollution studies, especially as part of a mosaic of species, allows for wide range comparisons of marine food chain contamination. PMID- 19290560 TI - Determination of total and available fractions of PAHs by SPME in oily wastewaters: overcoming interference from NAPL and NOM. AB - BACKGROUND, AIM, AND SCOPE: Polycyclic aromatic hydrocarbons (PAHs) are often found in oily wastewaters. Their presence is usually the result of human activities and has a negative effect on the environment. One important step in addressing this problem is to evaluate the effectiveness of PAH removal by biological processes since these are the most cost-effective treatments known today. Many techniques are presently available for PAH determination in wastewaters. Solid phase microextracion (SPME) is known to be one of the most effective techniques for this purpose. When analyzing complex matrices with substances such as natural organic matter (NOM) and non-aqueous phase liquids (NAPL), it is important to differentiate the free dissolved PAH from matrix bonded PAH. PAHs associated with the bonded fraction are less susceptible to biological treatment. The present study concerns the development of a simple and suitable methodology for the determination of the freely dissolved and the total fraction of PAHs present in oily wastewaters. The methodology was then applied to an oily wastewater from a fuel station retention basin. MATERIAL AND METHODS: Headspace SPME was used for analyzing PAH since the presence of a complex or dirty matrix in direct contact with the fiber may damage it. Four model PAHs anthracene, fluorene, phenanthrene, and pyrene-were analyzed by GC-MS. Negligible depletion SPME technique was used to determine the free fraction. Total PAH was determined by enhancing the mass transfer from the bonded phase to the freely dissolved phase by temperature optimization and the use of the method of standard additions. The PAH absorption kinetics were determined in order to define the optimal sampling conditions for this method. The fitting of the experimental data to a mathematical model was accomplished using Berkeley Madonna software. Humic acid and silicon oil were used as model NOM and NAPL, respectively, to study the effect of these compounds on the decrease of SPME response. Then, the method was evaluated with wastewater from a fuel station spill retention basin. RESULTS: The SPME kinetic parameters-k (1) (uptake rate), k (2) (desorption rate), and K (SPME) (partition coefficient)-were determined from experimental data modeling. The determination of the free fraction required 15-min sampling to ensure that PAH depletion from sample was below 1%. For total PAH, a 30-min extraction at 100 degrees C ensured the maximum signal response in the GC-MS. For the determination of free and total PAHs, extractions were performed before reaching the SPME equilibrium. The wastewater used in this study had no free fraction of the analyzed PAHs. However, the four studied PAHs were found when the method for total PAH was used. DISCUSSION: The addition of NOM and NAPL dramatically decreased the efficiency of the SPME. This decrease was the result of a greater partition of the PAHs to the NAPL and NOM phases. This fact was also observed in the analysis of the fuel station spill retention basin, where no free PAH was measured. However, using the method of standard addition for the determination of total PAH, it was possible to quantify all four PAHs. CONCLUSIONS: The method developed in the present study was found to be adequate to differentiate between free and total PAH present in oily wastewater. It was determined that the presence of NOM and NAPL had a negative effect on SPME efficiency. RECOMMENDATIONS AND PERSPECTIVES: The presence of binding substances had a great influence on SPME kinetics. Therefore, it is of extreme importance to determine their degree of interference when analyzing oily wastewaters or results can otherwise be erroneous. Other factors influencing the total PAH determinations should be considered in further studies. PMID- 19290561 TI - Realistic threshold policy with hysteresis to control predator-prey continuous dynamics. AB - This paper introduces a threshold policy with hysteresis (TPH) for the control of one-predator one-prey models. The models studied are the Lotka-Volterra and Rosenzweig-MacArthur two species density-dependent predator-prey models and the Arditi-Ginzburg nondimensional ratio-dependent model. The proposed policy (TPH) changes the dynamics of the system in such a way that a bounded oscillation is achieved confined to a region that does not allow extinction of either species. The policy can be designed by a suitable choice of so called virtual equilibrium points in a simple and intuitive manner. PMID- 19290562 TI - Suicide among Italian adolescents: 1970-2002. AB - The purpose of the present study was to analyze sex and regional differences in the suicide rate of adolescents and the methods they used for suicide in Italy during the period 1970-2002. Temporal trends and regional variations in suicide for Italian adolescents were retrieved from the Italian database on mortality for the period 1970-2002, collected by the Italian Census Bureau and processed by the Italian National Institute of Health-Statistics Unit. In the period 1970-2002, 3,069 adolescent suicides were monitored in Italy. Analyses of these suicides identified significant differences by region of residence and sex. Males were 2.1 times more likely than females to kill themselves. Male and female suicides had inverse trends in the years analyzed, so that the sex difference at the present time is the result of a continuous increase in male suicides and a decrease in female suicides since 1970. The dramatic peaks observed over the time period studied cannot be attributed to a single cause, indicating that further studies are needed to better understand the phenomenon. PMID- 19290563 TI - Genetic signature of rice domestication shown by a variety of genes. AB - Cultivated rice was domesticated from common wild rice. However, little is known about genetic adaptation under domestication. We investigated the nucleotide variation of both cultivated rice and its wild progenitors at 22 R-gene and 10 non-R-gene loci. A significant regression was observed between wild rice and rice cultivars in their polymorphic levels, particularly in their nonsynonymous substitutions (theta ( a )). Our data also showed that a similar proportion (approximately 60%) of nucleotide variation in wild rice was retained in cultivated rice in both R-genes and non-R-genes. Interestingly, the slope always was >1 and the intercept always >0 in linear regressions when a cultivar's polymorphism was x-axis. The slope and intercept values can provide a basis by which to estimate the founder effect and the strength of artificial direct selection. A larger founder effect than previously reported and a strong direct selection effect were shown in rice genes. In addition, two-directional selection was commonly found in differentiated genes between indica and japonica rice subspecies. This kind of selection may explain the mosaic origins of indica and japonica rice subspecies. Furthermore, in most R-genes, no significant differentiation between cultivated and wild rice was detected. We found evidence for genetic introgression from wild rice, which may have played an important role during the domestication of rice R-genes. PMID- 19290565 TI - Torsades de pointes in a child. PMID- 19290566 TI - Distribution of hybrid fungal symbionts and environmental stress. AB - Most asexual fungal symbionts of grasses in the genus Neotyphodium occurring in nature are of hybrid origin. Most hybrid Neotyphodium species result from interspecific hybridization events between pathogenic Epichloe species or co occurring non-hybrid Neotyphodium species. Current hypotheses for the prevalence of hybrid Neotyphodium species include reduction of mutation accumulation and increased adaptive response to environmental extremes. We tested the adaptive response hypothesis by characterizing the distribution of uninfected, hybrid, and non-hybrid Neotyphodium endophytes in 24 native Arizona fescue host populations and abiotic parameters at each locality. Infection was high in all host populations (>70%), but the majority of host populations were infected by non hybrid Neotyphodium (>50% on average). Principal component analysis indicates the frequency of plants infected with hybrid fungi is negatively related to soil nutrients and positively correlated with early spring moisture. Non-hybrid infected hosts are positively associated with soil nutrients and show a complex relationship with soil moisture (negative in early spring moisture, positive with late summer soil moisture). These results suggest the frequency of uninfected, hybrid, and non-hybrid infected plants is related to resource availability and abiotic stress factors. This supports the hypothesis that hybridization in asexual fungal symbionts increases host adaptability to extreme environments. PMID- 19290567 TI - Perceived effects of setting attributes on visitor experiences in wilderness: variation with situational context and visitor characteristics. AB - Understanding how setting attributes influence the nature of the visitor experience is crucial to effective recreation management. Highly influential attributes are useful indicators to monitor within a planning framework, such as Limits of Acceptable Change. This study sought to identify the setting attributes perceived to have the most profound effect on the ability to have "a real wilderness experience" and to assess the degree to which attribute importance varied with situational context and visitor characteristics. To this end, exiting hikers were surveyed at moderate and very high use trailheads in Alpine Lakes Wilderness, WA (USA), and Three Sisters Wilderness, OR (USA). They were asked about the degree to which encountering varying levels of different setting attributes would add to or detract from their experience. Attributes with the largest range of effect on experience, based on evaluations of different levels, were considered most important. The most influential attributes were litter and several types of campsite interaction--people walking through camp and number of other groups camping close by. The perceived importance of setting attributes did not vary much between wilderness locations with substantially different use levels, suggesting that conclusions are robust and generalizable across wilderness areas. There also was little difference in the perceptions of day and overnight visitors. In contrast, we found substantial variation in the perceived importance of setting attributes with variation in wilderness experience, knowledge, attachment, and motivation. Our results validate the emphasis of many wilderness management plans on indicators of social interaction, such as number of encounters. PMID- 19290568 TI - Accountability, reporting, or management improvement? Development of a state of the parks assessment system in New South Wales, Australia. AB - Management effectiveness evaluation has been recognized as an important mechanism for both reporting on and improving protected area management. The Convention on Biological Diversity's program of work on protected areas calls on all countries to implement such systems. In 2004, the first whole of system assessment of park management effectiveness, based on the IUCN-WCPA Management Effectiveness Evaluation Framework, was undertaken in New South Wales, Australia as part of a State of the Parks reporting requirement. This article describes the development of the State of the Parks assessment tool, its elements, and how it addresses the management effectiveness difficulties associated with assessments conducted across an extensive and diverse range of park types. The importance of engaging staff, at all levels, throughout the process is highlighted, as well as the adjustments made to the assessment tool based on staff feedback. While some results are presented, the main purpose of the article is to identify and discuss important procedural and methodological considerations. These include balancing quantitative and qualitative assessment approaches, achieving a comprehensive understanding of the management processes, and responding to any problems associated with assessments. PMID- 19290569 TI - Land use and land cover change analysis and prediction in the upper reaches of the Minjiang River, China. AB - Scientists have aimed at exploring land use and land cover change (LUCC) and modeling future landscape pattern in order to improve our understanding of the causes and consequences of these phenomena. This study addresses LUCC in the upper reaches of Minjiang River, China, from 1974 to 2000. Based on remotely sensed images, LUCC and landscape pattern change were assessed using cross tabulation and landscape metrics. Then, using the CLUE-S model, changes in area of four types of land cover were predicted for two scenarios considering forest polices over the next 20 years. Results showed that forestland decreased from 1974 to 2000 due to continuous deforestation, while grassland and shrubland increased correspondingly. At the same time, the farmland and settlement land increased dramatically. Landscape fragmentation in the study area accompanied these changes. Forestland, grassland, and farmland take opposite trajectories in the two scenarios, as does landscape fragmentation. LUCC has led to ecological consequences, such as biodiversity loss and lowering of ecological carrying capacity. PMID- 19290570 TI - Validation of the estimation of physiologic ability and surgical stress (E-PASS) score in liver surgery. AB - BACKGROUND: The estimation of physiologic ability and surgical stress (E-PASS) has been used to produce a numerical estimate of expected mortality and morbidity after elective gastrointestinal surgery. The aim of this study was to validate E PASS in a selected cohort of patients requiring liver resections (LR). METHODS: In this retrospective study, E-PASS predictor equations for morbidity and mortality were applied to the prospective data from 243 patients requiring LR. The observed rates were compared with predicted rates using Fisher's exact test. The discriminative capability of E-PASS was evaluated using receiver-operating characteristic (ROC) curve analysis. RESULTS: The observed and predicted overall mortality rates were both 3.3% and the morbidity rates were 31.3 and 26.9%, respectively. There was a significant difference in the comprehensive risk scores for deceased and surviving patients (p = 0.043). However, the scores for patients with or without complications were not significantly different (p = 0.120). Subsequent ROC curve analysis revealed a poor predictive accuracy for morbidity. CONCLUSIONS: The E-PASS score seems to effectively predict mortality in this specific group of patients but is a poor predictor of complications. A new modified logistic regression might be required for LR in order to better predict the postoperative outcome. PMID- 19290571 TI - Burden of surgical disease: does the literature reflect the scope of the international crisis? AB - BACKGROUND: Little is factually known about the burden of surgical disease and less is known about global surgical provision of care for diseases that may be treated, cured, or palliated by a surgical intervention. Despite the lack of information, surgical interventions are provided by a variety of agencies every day in the developing world. This literature review represents the first published comprehensive review of the global surgical literature. The primary goal was to collect and summarize what has been published on the current global burden of surgical disease and thereby encourage and promote the allocation of further research and resources. METHODS: A systematic review of English language publications on Pubmed or Medline was performed. RESULTS AND CONCLUSIONS: This report summarizes what little is known in terms of numeric estimates for the global burden of surgical disease. Globally, access and availability of surgical care in developing countries remains scarce, but the problem is receiving more attention for the first time in surgical circles. Much work remains in the effort to obtain reliable estimates of the global burden of surgical disease. PMID- 19290572 TI - Remedial operation for primary hyperparathyroidism. AB - BACKGROUND: Remedial surgery for patients with persistent or recurrent primary hyperparathyroidism (1 degrees HPT) remains a significant challenge. Cervical reexploration is technically difficult; reoperative neck anatomy is distorted by fibrosis and, as a result, remedial 1 degrees HPT patients carry an increased risk of injury to the recurrent (RLN) and superior laryngeal nerve(s) as well as to normal residual parathyroid tissue. Causative hyperfunctioning parathyroid tissue is also more frequently ectopic in the remedial setting and can thus be difficult to localize. METHODS: This report assimilates the current data underlying preoperative, intraoperative and postoperative remedial 1 degrees HPT management and presents an evidence-based algorithm for the management of remedial parathyroid disease. Recommendations are graded according to the quality of supporting data using the system initially developed by Sackett (Chest 95:2S 4S, 1989) and subsequently modified by Heinrich et al. (Ann Surg 243:154-168, 2006). RESULTS: Recent advances in preoperative localization and intraoperative adjuncts have lead to substantial improvements in outcomes after remedial surgery. Preoperative localization techniques, including sestamibi scintigraphy (MIBI), high resolution ultrasound (US), US-guided fine needle aspiration (FNA) and selective venous sampling (SVS), coupled with intraoperative adjuncts such as the rapid parathyroid hormone (PTH) assay have lead to reoperative cure rates as high as 96 percent. Nonetheless, management of remedial 1 degrees HPT varies significantly between surgeons and no formal recommendations standardizing the care of these patients have been published. CONCLUSIONS: Despite the significant challenges associated with remedial surgery for 1 degrees HPT, excellent outcomes can be reproducibly achieved when proper pre-, intra-, and postoperative management is employed. PMID- 19290573 TI - Establishing the evidence base for trauma quality improvement: a collaborative WHO-IATSIC review. AB - BACKGROUND: Quality improvement (QI) programs are an integral part of well developed trauma systems. However, they have not been extensively implemented globally. To promote greater use of effective QI programs, the World Health Organization (WHO) and the International Association for Trauma Surgery and Intensive Care (IATSIC) have been collaboratively developing the upcoming Guidelines for Trauma Quality Improvement Programmes. As part of the development of this publication and to satisfy global demands for WHO guidelines to be evidence based, we conducted a thorough literature search on the effectiveness of trauma QI programs. METHODS: The review was based on a PubMed search of all articles reporting an outcome from a trauma QI program. RESULTS: Thirty-six articles were identified that reported results of evaluations of a trauma QI program or in which the trauma QI program was integrally related to identification and correction of specific problems. Thirteen of these articles reported on mortality as their main outcome; 12 reported on changes in morbidity (infection rates, complications), patient satisfaction, costs, or other outcomes of tangible patient benefit; and 11 reported on changes in process of care. Thirty articles addressed hospital-based care; four system-wide care; and two prehospital care. Thirty-four articles reported an improvement in the outcome assessed; two reported no change; and none reported worsening of the outcome. Five articles also reported cost savings. CONCLUSIONS: Trauma QI programs are consistently shown to improve the process of care, decrease mortality, and decrease costs. Further efforts to promote trauma QI globally are warranted. These findings support the further development and promulgation of the WHO-IATSIC Guidelines for Trauma QI Programmes. PMID- 19290574 TI - Investigation of the hemodynamic effect of stent wires on renal arteries in patients with abdominal aortic aneurysms treated with suprarenal stent-grafts. AB - The purpose of the study was to investigate the hemodynamic effect of stent struts (wires) on renal arteries in patients with abdominal aortic aneurysms (AAAs) treated with suprarenal stent-grafts. Two sample patients with AAA undergoing multislice CT angiography pre- and postsuprarenal fixation of stent grafts were selected for inclusion in the study. Eight juxtarenal models focusing on the renal arteries were generated from the multislice CT datasets. Four types of configurations of stent wires crossing the renal artery ostium were simulated in the segmented aorta models: a single wire crossing centrally, a single wire crossing peripherally, a V-shaped wire crossing centrally, and multiple wires crossing peripherally. The blood flow pattern, flow velocity, wall pressure, and wall shear stress at the renal arteries pre- and post-stent-grafting were analyzed and compared using a two-way fluid structure interaction analysis. The stent wire thickness was simulated with a diameter of 0.4, 1.0, and 2.0 mm, and hemodynamic analysis was performed at different cardiac cycles. The interference of stent wires with renal blood flow was mainly determined by the thickness of stent wires and the type of configuration of stent wires crossing the renal ostium. The flow velocity was reduced by 20-30% in most of the situations when the stent wire thickness increased to 1.0 and 2.0 mm. Of the four types of configuration, the single wire crossing centrally resulted in the highest reduction of flow velocity, ranging from 21% to 28.9% among three different wire thicknesses. Wall shear stress was also dependent on the wire thickness, which decreased significantly when the wire thickness reached 1.0 and 2.0 mm. In conclusion, our preliminary study showed that the hemodynamic effect of suprarenal stent wires in patients with AAA treated with suprarenal stent-grafts was determined by the thickness of suprarenal stent wires. Research findings in our study are useful for follow-up of patients treated with suprarenal stent grafts to ensure long-term safety of the suprarenal fixation. PMID- 19290575 TI - Endovascular treatment of a ruptured profunda femoral artery branch after fogarty thrombectomy of a femoro-femoral crossover arterial graft: a case report and review of the literature. AB - We present a very rare case of a life-threatening rupture of a profunda femoral artery distal branch after a Fogarty thrombectomy of a thrombosed crossover synthetic graft between the ipsilateral common femoral artery and a contralateral iliac-popliteal graft; the bleeding profunda femoral artery branch was successfully embolized with metallic coils through the axillary artery approach. PMID- 19290577 TI - Occlusion balloon catheter-assisted removal of a wedged catheter fragment without a free end from a pulmonary artery. AB - Inadvertent embolization of guide wires and catheter fragments are relatively common due to the widespread usage of central venous catheters. A variety of percutaneous techniques has been proposed to retrieve intravascular foreign bodies. The majority of described methods have been dependent on a free end or exposed portion of the foreign body to grasp, snare, or hook. We report the simple use of a balloon catheter to relocate a catheter fragment without a free end that was wedged into a small pulmonary branch to a larger branch. In the larger branch, the fragment was then easily engaged with a standard snare device. PMID- 19290578 TI - Dysphagia following a total laryngectomy: the effect on quality of life, functioning, and psychological well-being. AB - The aim of this study was to investigate the effect that dysphagia has on quality of life (QoL), functioning, and psychological well-being of people who have undergone a total laryngectomy. A questionnaire battery was sent to all members (N = 197) of the Laryngectomee Association of NSW, Australia. QoL and functioning were assessed using the World Health Organisation Quality of Life-Bref (WHOQoL Bref) and the University of Washington QoL (UW-QoL) measures. Psychological well being was measured using the Depression Anxiety and Stress Scale (DASS). One hundred ten questionnaires (56%) were completed and returned. There were no significant differences in QoL, as measured by the WHOQoL-Bref, between those laryngectomees with and without dysphagia. Laryngectomees with dysphagia, however, had significantly impaired functioning and markedly reduced social participation as measured by the UW-QoL. Significantly higher levels of depression and anxiety were also documented in those laryngectomees who had dysphagia. Dysphagia may not necessarily determine QoL following a total laryngectomy. However, it may have a negative impact on functioning and on psychological well-being. PMID- 19290579 TI - Applying a presentation content manifest for signing clinical documents. AB - In this paper, we demonstrate how to digitally sign a content manifest of a presentable clinical document that contains multiple clinical data with presentations. Only one signature is needed for an entire clinical document with multiple data resources, which can reduce the computation time during signing and verifying processes. In the radiology field, a report may contain text descriptions, images, and annotations that are stored separately in different data resources. The manifest signature would be a proper means for integrity checking for all the clinical data within the manifest. The manifest signature can be extended with a trusted third party to add a digital time signature for long-term verifiability. The performance of the manifest signing compared with that of a traditional digital signing was evaluated. The new manifest signature can be used for signing different types of presentable clinical documents, such HL7 CDA documents and DICOM image reports. PMID- 19290581 TI - Development of transdisciplinarity among students placed with a sustainability for health research project. AB - Transdisciplinary education on sustainability for health has been primarily developed in high-income countries, yet the need in countries with limited research and human resource investments remains urgent. Little empiric documentation of the facilitators and barriers to transdisciplinary learning in such countries has been described. We assessed transdisciplinary learning among students of different disciplines collaborating with an Ecuadorian sustainability for health research project. Six undergraduate students from four different disciplinary backgrounds were incorporated through work-study agreements with provincial university academic supervisors. Learning was fostered and monitored through participant observations by a field supervisor. Students' learning was evaluated through subsequent in-depth interviews and visualization methods. Academic supervisor key informant and co-investigator observations aided triangulation. Qualitative data were analyzed using indicators of transdisciplinary thinking. Principal factors facilitating transdisciplinary learning were interaction with social actors, the integration of work with other disciplines, the use of alternative research techniques and methods, and the constant support of the field supervisor. Inhibiting factors included the existence of rigid academic rules, lack of training of the academic supervisors in diverse research methods, and social pressures to implement unidisciplinary foci. At the end of their link with the project, students had developed both cognitive outcomes and attitudinal values relevant to sustainable development for health. In countries with limited investments in research and human resources development, transdisciplinary approaches with social actors and engaged researchers can sensitize new professionals training in traditional academic contexts to the ecological-social-health problems faced by poor majorities and encourage their subsequent work on sustainability for human health. PMID- 19290582 TI - Recent advances toward the clinical application of PTH (1-34) in fracture healing. AB - PTH 1-34, an active form of parathyroid hormone, has been shown to enhance osteoblastic bone formation when administered as a daily subcutaneous injection. The effect of the intermittent administration of PTH (1-34) is an uncoupling of bone turnover with an increase in bone mass and density and decrease in risk of vertebral and nonvertebral fractures. While PTH (1-34) has been used clinically to increase bone mass and reduce fracture risk in postmenopausal women with osteoporosis, there is increasing evidence that PTH (1-34) may promote fracture healing. Animal studies have demonstrated accelerated callus formation with enhanced remodeling and biomechanical properties of the healing fracture. Given these effects, PTH (1-34) will likely be used clinically to enhance fracture union in poor healing situations such as osteoporosis and recalcitrant nonunions. PMID- 19290583 TI - A musculoskeletal profile of elite female soccer players. AB - The purpose of this study is to identify lower-extremity (LE) musculoskeletal characteristics of elite female soccer players and to determine whether differences between dominant and nondominant extremities exist with respect to strength, flexibility, and range of motion. Physical data were collected from 26 female professional soccer players. Core control, hip and knee passive range of motion (PROM), LE flexibility, hip abductor strength, and dynamic functional alignment were assessed for each LE. Of 26 subjects, 21 scored 2/5 or less on core control. Mean hip internal rotation and external rotation were 33 degrees (+/-8 degrees) and 25 degrees (+/-6.7 degrees), respectively. All subjects had shortened two-joint hip flexors with an average knee flexion angle of 50 degrees (+/-11 degrees) and increased femoral anteversion. Forty one of 48 dominant limbs and 42 of 48 nondominant limbs demonstrated deviations from neutral alignment during step down or single-leg squat. Of 25 subjects, 15 demonstrated a stiff knee landing and/or takeoff. All subjects demonstrated limitations in hip external rotation PROM and hip flexor length. There was no difference between dominant and nondominant LEs in all variables including hip abductor strength. Additional research is needed to determine if there is a correlation between the musculoskeletal characteristics, LE biomechanics, and potential risk for injury. LEVEL OF EVIDENCE: IV. PMID- 19290584 TI - Exploration of the dynamical evolution and the associated energetics of water nanoclusters formed in a hydrophobic solvent. AB - Exploration of the intermolecular binding energy in nanometer-sized small water clusters in hydrophobic solvents and its evolution with the increase in the cluster size until bulk-type geometry is reached constitute a fascinating area of research in contemporary chemical/biological physics. In this contribution we have used femtosecond/picosecond-resolved solvation dynamics and fluorescence anisotropy techniques to explore the dynamical evolution of water clusters in dioxane continuum as a function of water concentration. We have also used temperature dependent picosecond-resolved solvation dynamics in order to explore the magnitude of the intermolecular bonding energy in the water clusters in bulk dioxane. PMID- 19290585 TI - Description of bound reactive dynamics within the approximate quantum trajectory framework. AB - The quantum trajectory framework incorporates quantum effects on dynamics through the quantum potential acting on a trajectory ensemble in addition to the classical potential. A global quadratic approximation to the quantum potential makes the method practical in many dimensions and captures dominant quantum effects in semiclassical systems. In this paper the approach is further developed to describe the "double well" dynamicsa prototype of the proton transfer reactionswhich exhibits the "hard" quantum effect of tunneling. Accurate description is achieved by combining the approximate quantum trajectory dynamics with the population amplitudes in the reactant and product wells. The quantum trajectory dynamics is defined by the asymptotic classical potentials. The population amplitudes represented in a small basis describe transfer between the wells. The method is exact if the reactant/product potentials are quadratic and the basis size is sufficiently large. In the semiclassical regime the trajectory dynamics is approximate and the basis size can be as small as two functions. The approach is fully compatible with the trajectory description of multidimensional systems capturing quantum tunneling along the reactive coordinate and zero-point energy flow among all degrees of freedom. PMID- 19290586 TI - Infrared spectroscopy and structures of cobalt carbonyl cations, Co(CO)n+ (n = 1 9). AB - Cobalt carbonyl cations of the form Co(CO)(n)(+) (n = 1-9) are produced in a molecular beam by laser vaporization in a pulsed nozzle source. These ions, and their corresponding "argon-tagged" analogues, Co(CO)(n)(Ar)(m)(+), are studied with mass-selected infrared photodissociation spectroscopy in the carbonyl stretching region. The number of infrared-active bands, their frequency positions, and their relative intensities provide distinctive patterns allowing determination of the geometries and electronic structures of these complexes. Co(CO)(5)(+) has a completed coordination sphere, consistent with its expected 18 electron stability, and it has the same structure (D(3h) trigonal bipyramid) as its neutral isoelectronic analog Fe(CO)(5). The carbonyl stretches in Co(CO)(5)(+) are less red-shifted than those in Fe(CO)(5) because of charge induced reduction in the pi back-bonding. Co(CO)(1-4)(+) complexes have triplet ground states, but the spin changes to a singlet for the Co(CO)(5)(+) complex. PMID- 19290587 TI - Theoretical investigations of mode mixing in vibrationally excited states of CH5+. AB - Using diffusion Monte Carlo, five vibrational excited states of CH(5)(+) and CD(5)(+) are evaluated and analyzed. Here, we focus on the fundamentals in the five modes that are generated by requiring that the wave functions change sign at specified values of the five symmetry-adapted linear combinations (SALCs) of the CH or CD bond lengths. Even though the definitions of these modes are based on displacements of the CH or CD bond lengths, the frequencies are found to be low compared to previously calculated CH vibrational frequencies in this molecule. The totally symmetric mode, with A(1)(+) symmetry, has a calculated frequency of 2164 and 1551 cm(-1) for CH(5)(+) and CD(5)(+). The frequencies of the four fundamentals that arise from excitation of the four SALCs that transform under G(1)(+) symmetry have frequencies that range from 1039 to 1383 cm(-1) and from 628 to 893 cm(-1) in CH(5)(+) and CD(5)(+), respectively. The origins of the broken degeneracy are investigated and explained to reflect extensive coupling to the two low-frequency modes that lead to isomerization of CH(5)(+). PMID- 19290588 TI - Surface-enhanced Raman spectroscopy of benzenethiol adsorbed from the gas phase onto silver film over nanosphere surfaces: determination of the sticking probability and detection limit time. AB - A chemical warfare agent (CWA) gas detector based on surface-enhanced Raman spectroscopy (SERS) using robust nanostructured substrates and a portable Raman spectrometer is a promising alternative to existing modalities. A gas-dosing apparatus was constructed to simulate chemical gas exposure and provide a platform for quantitative analysis of SERS detection. As a first step toward characterizing SERS detection from the gas phase, benzenethiol (BT) has been chosen as the test analyte. SERS spectra were monitored during BT adsorption onto a silver film over a nanosphere (AgFON) substrate. The SERS detection limit time (DLt) for BT on a AgFON at 356 K is found to be 6 ppm-s (30 mg-s m(-3)) for a data acquisition time (t(acq)) of 1 s. The DLt for this kinetically controlled sensor is fundamentally determined by the low sticking probability of BT on AgFONs which is determined to be approximately 2 x 10(-5) at 356 K. The sticking probability increases with increasing temperature consistent with an adsorption activation barrier of approximately 13 kJ mol(-1). Although the DLts found in the present study for BT are in the low ppm-s, a theoretical model of SERS detection indicates DLts below 1 ppb s(-1) for t(acq)= 1 s are, in fact, achievable using existing portable Raman instrumentation and AgFON surfaces. Achieving this goal requires the sticking probability be increased 3 orders of magnitude, illuminating the importance of appropriate surface functionalization. PMID- 19290589 TI - Midinfrared extinction spectra of submicron carbohydrate particles generated by a pneumatic atomizer. AB - Carbohydrate aqueous solutions of trehalose, glucose, and fructose were sprayed by using a pneumatic atomizer, and their infrared extinction spectra were recorded for the region from 700 to 5000 cm(-1). Analysis based on Mie scattering theory indicated that sprayed droplets transformed into nonvolatile amorphous nanoparticles by solvent evaporation. Average diameters of these particles were estimated to be about 100 nm, which was further confirmed by differential mobility analysis. The results demonstrate that nanoparticles can be created by spray drying of aqueous solutions, and that the sizes, phases, and structures of these particles can be well characterized by infrared extinction spectroscopy. PMID- 19290590 TI - Optical properties of anisotropic core-shell pyramidal particles. AB - This paper describes an approach to fabricate anisotropic core-shell particles by assembling dielectric beads within fabricated noble metal pyramidal structures. Particles with gold (Au) shells and different dielectric cores were generated, and their optical properties were characterized by single particle spectroscopy. Because of their unique geometry, these particles exhibit multiple plasmon resonances from visible to near-IR wavelengths. PMID- 19290591 TI - Effects of alkali cations and halide anions on the DOPC lipid membrane. AB - By means of molecular dynamics simulations with an all-atom force field, we investigated the affinities of alkali cations and halide anions for the dioleoylphosphatidylcholine lipid membrane in aqueous salt solutions. In addition, changes in phospholipid lateral diffusion and in headgroup mobility upon adding NaCl were observed using fluorescence spectroscopy. The simulations revealed that sodium is attracted to the headgroup region with its concentration being maximal in the vicinity of the phosphate groups. Potassium and cesium, however, do not preferentially adsorb to the membrane. Similarly, halide anions do not exhibit a strong affinity for the lipid headgroups but merely compensate for the positive charge of the sodium countercations. Nevertheless, larger halides such as bromide and iodide penetrate deeper into the headgroup region toward the boundary with the hydrophobic alkyl chain, this effect being likely underestimated within the present nonpolarizable force field. Addition of alkali halide salts modifies physical properties of the bilayer including the electronic density profiles, the electrostatic potential, and the area per lipid headgroup. PMID- 19290592 TI - Surface-enhanced vibrational raman optical activity: a time-dependent density functional theory approach. AB - In this work we present the first simulations of the surface-enhanced Raman optical activity (SEROA) using time-dependent density functional theory (TDDFT). A consistent treatment of both the chemical and electromagnetic enhancements is achieved by employing a recently developed approach based on a short-time approximation for the Raman and ROA cross-sections. As an initial application we study a model system consisting of adenine interacting with a Ag(20) cluster. Because both the silver cluster and adenine in the absorption geometry are achiral, the chiroptical properties are due to the interactions between the two systems. Our results show that the total enhancement is on the order of 10(4) both for SEROA and SERS. However, the chemical enhancement is found to be larger for SEROA than for SERS. The results presented here show that SEROA can be a useful approach for studying induced chirality in small metal clusters due to the absorption of molecules. PMID- 19290593 TI - Allopregnanolone (3alpha-hydroxy-5alpha-pregnan-20-one) derivatives with a polar chain in position 16alpha: synthesis and activity. AB - The lipophilic nature of allopregnanolone prevents its user-friendly application in human medicine. On inspiration by previously prepared allopregnanolone with a 16alpha-bound tetraethylammonium salt, an attempt was made to produce allopregnanolone analogues with polar groups introduced into position 16alpha with the goal of increasing water solubility, brain accessibility, and potency of neuroactive steroids. The Michael addition to derivatives of pregn-16-en-20-one was the key reaction step. The link between the steroid skeleton and the new side chain was either a methylene group (when diethyl malonate was added) or an oxygen atom (when a hydroxy derivative was added). [(35)S]TBPS displacement was used to evaluate the products. Several carbamates (but not their parent alcohols) displaced TBPS from the picrotoxin binding site on GABA(A) receptors. Although none of them was more potent than the above ammonium salt, which stimulated this study, their nonionic nature should not prevent their passage into the brain. PMID- 19290594 TI - Comparison of N-terminal modifications on neurotensin(8-13) analogues correlates peptide stability but not binding affinity with in vivo efficacy. AB - Neurotensin(8-13) and two related analogues were used as model systems to directly compare various N-terminal peptide modifications representing both commonly used and novel capping groups. Each N-terminal modification prevented aminopeptidase cleavage but surprisingly differed in its ability to inhibit cleavage at other sites, a phenomenon attributed to long-range conformational effects. None of the capping groups were inherently detrimental to human neurotensin receptor 1 (hNTR1) binding affinity or receptor agonism. Although the most stable peptides exhibited the lowest binding affinities and were the least potent receptor agonists, they produced the largest in vivo effects. Of the parameters studied only stability significantly correlated with in vivo efficacy, demonstrating that a reduction in binding affinity at NTR1 can be countered by increased in vivo stability. PMID- 19290595 TI - Backbone amide linker in solid-phase synthesis. PMID- 19290596 TI - Importance of side chains and backbone length in defect modeling of poly(3 alkylthiophenes). AB - Geometric defects in conjugated polymers play a critical role in determining electronic structure and properties such as charge carrier mobility and band gap. Because the relative roles of individual defects are experimentally difficult to discern, computational approaches provide valuable insight if appropriate molecular models are used. Poly(3-alkylthiophenes) are often modeled with very short backbones and without their side chains. We demonstrate the shortcomings of this approach for modeling torsional disorder in poly(3-hexylthiophene) (P3HT). Using a hybrid density functional model, we identify a minimal acceptable model to comprise approximately 10 monomers with explicitly treated alkane side chains. Potential applications of this work extend to polymer electronics and optoelectronics. PMID- 19290597 TI - Ab initio potential energy surfaces for the ground (X1A') and excited (A1A'') electronic states of HGeBr and the Absorption and emission spectra of HGeBr/DGeBr. AB - We report global potential energy surfaces for both the ground (X(1)A') and the excited (A(1)A'') electronic states of HGeBr as well as the transition dipole moment surface between them using an internally contracted multireference configuration interaction method with the Davidson correction and an augmented correlation-consistent polarized valence quadruple-zeta basis set. Vibrational energy levels of HGeBr and DGeBr are calculated on both the ground and the excited electronic states and found in good agreement with the available experimental band origins. In addition, the A(1)A''-X(1)A' absorption and emission spectra of the two isotopomers were obtained, and an excellent agreement with the available experimental spectra was found. PMID- 19290598 TI - Volatile compounds in acacia, chestnut, cherry, ash, and oak woods, with a view to their use in cooperage. AB - Extracts of wood from acacia, European ash, American ash, chestnut, cherry, and three oak species (Quercus pyrenaica, Quercus alba and Quercus petraea) before and after toasting in cooperage were studied by GC-MS. 110 compounds were detected, and 97 of them were identified. In general, all studied woods showed more lignin derivatives than lipid and carbohydrate derivatives, with a higher variety of compounds detected and abundance of them. The toasting led to an increase in the concentrations of most of these compounds, and this increase is especially important in acacia, chestnut and ash woods. The cis and trans isomers of beta-methyl-gamma-octalactone and isobutyrovanillone were only detected in oak wood, 3,4-dimethoxyphenol and 2,4-dihydroxybenzaldehyde only in acacia wood, and p-anisaldehyde and benzylsalicylate only in cherry wood, before and after toasting, and these compounds could be considered chemical markers for each one of these woods. Moreover, each wood has a characteristic volatile composition, from a quantitative point of view, and therefore we can expect a characteristic sensorial profile. The oak wood turned out to be the most balanced, since although it provides a lot of volatile compounds to the aroma and flavor of aged wine, it can do so without masking their primary and secondary aroma. On the whole, toasted acacia and chestnut woods showed a very high richness of studied compounds, as lignin as lipid and carbohydrate derivatives, while cherry and ash were much richer than toasted oak wood in lignin derivatives, but much poorer in lipid and carbohydrate derivatives. PMID- 19290599 TI - Strong H...F hydrogen bonds as synthons in polymeric quantum magnets: structural, magnetic, and theoretical characterization of [Cu(HF2)(pyrazine)2]SbF6, [Cu2F(HF)(HF2)(pyrazine)4](SbF6)2, and [CuAg(H3F4)(pyrazine)5](SbF6)2. AB - Three Cu(2+)-containing coordination polymers were synthesized and characterized by experimental (X-ray diffraction, magnetic susceptibility, pulsed-field magnetization, heat capacity, and muon-spin relaxation) and electronic structure studies (quantum Monte Carlo simulations and density functional theory calculations). [Cu(HF(2))(pyz)(2)]SbF(6) (pyz = pyrazine) (1a), [Cu(2)F(HF)(HF(2))(pyz)(4)](SbF(6))(2) (1b), and [CuAg(H(3)F(4))(pyz)(5)](SbF(6))(2) (2) crystallize in either tetragonal or orthorhombic space groups; their structures consist of 2D square layers of [M(pyz)(2)](n+) that are linked in the third dimension by either HF(2)(-) (1a and 1b) or H(3)F(4)(-) (2). The resulting 3D frameworks contain charge-balancing SbF(6)(-) anions in every void. Compound 1b is a defective polymorph of 1a, with the difference being that 50% of the HF(2)(-) links are broken in the former, which leads to a cooperative Jahn-Teller distortion and d(x(2))(-y(2)) orbital ordering. Magnetic data for 1a and 1b reveal broad maxima in chi at 12.5 and 2.6 K and long-range magnetic order below 4.3 and 1.7 K, respectively, while 2 displays negligible spin interactions owing to long and disrupted superexchange pathways. The isothermal magnetization, M(B), for 1a and 1b measured at 0.5 K reveals contrasting behaviors: 1a exhibits a concave shape as B increases to a saturation field, B(c), of 37.6 T, whereas 1b presents an unusual two-step saturation in which M(B) is convex until it reaches a step near 10.8 T and then becomes concave until saturation is reached at 15.8 T. The step occurs at two thirds of M(sat), suggesting the presence of a ferrimagnetic structure. Compound 2 shows unusual hysteresis in M(B) at low temperature, although chi vs T does not reveal the presence of a magnetic phase transition. Quantum Monte Carlo simulations based on an anisotropic cubic lattice were applied to the magnetic data of 1a to afford g = 2.14, J = -13.4 K (Cu-pyz-Cu), and J(perpendicular) = 0.20 K (Cu-F...H...F-Cu), while chi vs T for 1b could be well reproduced by a spin-1/2 Heisenberg uniform chain model for g = 2.127(1), J(1) = -3.81(1), and zJ(2) = -0.48(1) K, where J(1) and J(2) are the intra- and interchain exchange couplings, respectively, which considers the number of magnetic nearest-neighbors (z). The M(B) data for 1b could not be satisfactorily explained by the chain model, suggesting a more complex magnetic structure in the ordered state and the need for additional terms in the spin Hamiltonian. The observed variation in magnetic behaviors is driven by differences in the H...F hydrogen-bonding motifs. PMID- 19290600 TI - Confocal Raman microscopy of the interfacial regions of liquid chromatographic stationary-phase materials. AB - The influence of organic modifiers on the structure of reversed-phase liquid chromatographic (RPLC) stationary phases has been a topic of considerable investigation. Retention of organic modifiers in the stationary phase has previously been determined by chromatographic measurements, and the polarity and heterogeneity of the resulting solvation environment has been studied using solvatochromic, fluorescent, and spin probes. In the present work, the composition and solvation environment of a stationary phase is investigated using confocal Raman microscopy, which allows the in situ examination of the solvation environment within individual chromatographic stationary phase particles without the use of probe molecules. The accumulation of organic modifiers in the stationary phase can be quantified, and the environment of the modifier in the stationary phase can be determined. Specifically, we have investigated the interactions of acetonitrile with C(18)-functionalized silica particles using confocal Raman microscopy, which enables the sampling of small (approximately 1 fL) volumes within individual 10 microm particles. Bare chromatographic silica was also studied in order to investigate the interactions of acetonitrile with surface silanols. The nitrile-stretching (nu(CN)) frequency of acetonitrile responds sensitively to the dipolarity of its local microenvironment. The populations of solution-phase and interfacial acetonitrile are thus spectroscopically distinguishable. Scattering from nu(CN) shows contributions from three different environments within a single RPLC chromatographic particle: acetonitrile in the interparticle mobile phase, C(18)-chain associated acetonitrile, and acetonitrile that is interacting with residual surface silanols. Data are presented quantifying these populations and characterizing their environments within single stationary phase particles. PMID- 19290601 TI - Targeted N-linked glycosylation analysis of H5N1 influenza hemagglutinin by selective sample preparation and liquid chromatography/tandem mass spectrometry. AB - Using liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis of deglycosylated and intact glycopeptides from tryptic digests of whole influenza virus, we determined that the six predicted N-linked glycosylation sites within the N-terminal ectodomain of hemagglutinin (HA) from three selected H5N1 strains are occupied. The use of selective sample preparation strategies, including solid phase extraction (SPE) of glycopeptides via hydrazide capture chemistry as well as hydrophilic interaction liquid chromatography (HILIC), sufficiently reduced sample complexity to allow determination of occupied glycosylation sites. The specific amino acid sequence of the tryptic glycopeptides for the identified sites varied slightly among strains, but the overall locations of the occupied glycosylation sites were conserved in the protein sequence. We used this knowledge of glycosylation site occupation to examine the glycans attached to these occupied sites on HA for a reassortant H5N1 strain grown in embryonated chicken eggs. By applying mass spectrometry-based methodologies for examining glycosylation to the study of influenza virus proteins, we can better understand the effect that this post-translational modification has upon the virulence and antigenicity of emerging strains. PMID- 19290602 TI - Rapid proton conduction through unfreezable and bound water in a wholly aromatic pore-filling electrolyte membrane. AB - We found that protons rapidly conduct through unfreezable and bound water in a pore-filling electrolyte membrane (PF-membrane), although many ions usually conduct through free water contained in polymer electrolytes. PF-membrane is a unique membrane that can suppress the swelling of filled sulfonated poly(arylene ether sulfone) (SPES) because of its rigid polyimide substrate. Based on low temperature DSC measurements, this strong suppression of swelling resulting from the special structure of the polymer electrolyte results in unfreezable and bound water only; it does not contain any free water. Protons rapidly conduct through this structure. In addition, the activation energy of the proton conduction decreased from 16.3 to 9.1 kJ/mol in proportion to the increase in the ion exchange capacity (IEC) of the filled SPES, unlike the almost constant values of the SPES-cast membranes. This tendency of PF-membrane occurred because of the structure of the membrane, where the concentration of the sulfonic acid groups increased with increase in IEC, which became possible by squeezing free water using the swelling suppression of filled SPES. Without being constrained by the PF-membrane, this unique proton conduction through the structured water and highly concentrated sulfonic acid groups will help to develop future polymer electrolytes, particularly in the fuel cell field where protons need to conduct at various conditions such as temperatures below 0 degrees C, combined high temperature and low humidity, and the presence of fuels. PMID- 19290603 TI - Configuration-driven unitary group approach for generalized Van Vleck variant multireference perturbation theory. AB - A new, efficient, configuration-driven algorithm utilizing the unitary group approach (UGA) was developed and implemented for the generalized van Vleck perturbation theory (GVVPT) variant of multireference perturbation theory. The computational speed has been improved by 1 or 2 orders of magnitude compared to the previous implementation based on the Table-CI technique. It is shown that the reformulation is applicable to both the second-order (GVVPT2) and third-order (GVVPT3) approximations. Calculations on model problems and on a chemically realistic description of cyclobutadiene are used to illustrate the performance of the method. The calculations on cyclobutadiene, using over 2.3 billion CSFs, provide results on geometric parameters and the barrier height of the automerization reaction in good agreement with established high accuracy results. PMID- 19290604 TI - Functional representation for the born-oppenheimer diagonal correction and born huang adiabatic potential energy surfaces for isotopomers of H3. AB - Multireference configuration interaction (MRCI) calculations of the Born Oppenheimer diagonal correction (BODC) for H(3) were performed at 1397 symmetry unique configurations using the Handy-Yamaguchi-Schaefer approach; isotopic substitution leads to 4041 symmetry-unique configurations for the DH(2) mass combination. These results were then fit to a functional form that permits calculation of the BODC for any combination of isotopes. Mean unsigned fitting errors on a test grid of configurations not included in the fitting process were 0.14, 0.12, and 0.65 cm(-1) for the H(3), DH(2), and MuH(2) isotopomers, respectively. This representation can be combined with any Born-Oppenheimer potential energy surface (PES) to yield Born-Huang (BH) PESs; herein, we choose the CCI potential energy surface, the uncertainties of which ( approximately 0.01 kcal/mol) are much smaller than the magnitude of the BODC. Fortran routines to evaluate these BH surfaces are provided. Variational transition state theory calculations are presented comparing thermal rate constants for reactions on the BO and BH surfaces to provide an initial estimate of the significance of the diagonal correction for the dynamics. PMID- 19290605 TI - Cyclohexane isomerization. Unimolecular dynamics of the twist-boat intermediate. AB - Direct dynamics simulations were performed at the HF/6-31G level of theory to investigate the intramolecular and unimolecuar dynamics of the twist-boat (TB) intermediate on the cyclohexane potential energy surface (PES). Additional calculations were performed at the MP2/aug-cc-pVDZ level of theory to further characterize the PES's stationary points. The trajectories were initiated at the C(1) and C(2) half-chair transition states (TSs) connecting a chair conformer with a TB intermediate, via an intrinsic reaction coordinate (IRC). Energy was added in accord with a microcanonical ensemble at the average energy for experiments at 263 K. Important nontransition state theory (TST), non-IRC, and non-RRKM dynamics were observed in the simulations. Trajectories initially directed toward the chair conformer had a high probability of recrossing the TS, with approximately 30% forming a TB intermediate instead of accessing the potential energy well for the conformer. The TB intermediate initially formed was not necessarily the one connected to the TS via the IRC. Of the trajectories initiated at the C(2) half-chair TS and initially directed toward the chair conformer, 35% formed a TB intermediate instead of the chair conformer. Also, of the trajectories forming a TB intermediate, only 16% formed the TB intermediate connected with the C(2) TS via the IRC. Up to eight consecutive TB --> TB isomerizations were followed, and non-RRKM behavior was observed in their dynamics. A TB can isomerize to two different TBs, one by a clockwise rotation of C-C-C-C dihedral angles and the other by a counterclockwise rotation. In contrast to RRKM theory, which predicts equivalent probabilities for these rotations, the trajectory dynamics show they are not equivalent and depend on whether the C(1) or C(2) half-chair TS is initially excited. Non-RRKM dynamics is also observed in the isomerization of the TB intermediates to the chair conformers. RRKM theory assumes equivalent probabilities for isomerizing to the two chair conformers. In contrast, for the first and following TB intermediate formed, there is a preference to isomerize to the chair conformer connected to the TS at which the trajectories were initiated. For the first TB intermediate formed, approximately 30% of the isomerization is to a chair conformer, but this fraction decreases for the later formed TB intermediates and becomes approximately 10% for the eighth consecutive TB intermediate formed. PMID- 19290606 TI - Improved methods for Feynman path integral calculations of vibrational-rotational free energies and application to isotopic fractionation of hydrated chloride ions. AB - We present two enhancements to our methods for calculating vibrational-rotational free energies by Feynman path integrals, namely, a sequential sectioning scheme for efficiently generating random free-particle paths and a stratified sampling scheme that uses the energy of the path centroids. These improved methods are used with three interaction potentials to calculate equilibrium constants for the fractionation behavior of Cl(-) hydration in the presence of a gas-phase mixture of H(2)O, D(2)O, and HDO. Ion cyclotron resonance experiments indicate that the equilibrium constant, K(eq), for the reaction Cl(H(2)O)(-) + D(2)O right harpoon over left harpoon Cl(D(2)O)(-) + H(2)O is 0.76, whereas the three theoretical predictions are 0.946, 0.979, and 1.20. Similarly, the experimental K(eq) for the Cl(H(2)O)(-) + HDO right harpoon over left harpoon Cl(HDO)(-) + H(2)O reaction is 0.64 as compared to theoretical values of 0.972, 0.998, and 1.10. Although Cl(H(2)O)(-) has a large degree of anharmonicity, K(eq) values calculated with the harmonic oscillator rigid rotator (HORR) approximation agree with the accurate treatment to within better than 2% in all cases. Results of a variety of electronic structure calculations, including coupled cluster and multireference configuration interaction calculations, with either the HORR approximation or with anharmonicity estimated via second-order vibrational perturbation theory, all agree well with the equilibrium constants obtained from the analytical surfaces. PMID- 19290607 TI - Confinement spectroscopy: probing single DNA molecules with tapered nanochannels. AB - We demonstrate a confinement spectroscopy technique capable of probing small conformational changes of unanchored single DNA molecules in a manner analogous to force spectroscopy, in the regime corresponding to femtonewton forces. In contrast to force spectroscopy, various structural forms of DNA can easily be probed, as indicated by experiments on linear and circular DNA. The extension of circular DNA is found to scale according to the de Gennes exponent, unlike for linear DNA. PMID- 19290608 TI - Raman spectroscopy of graphene edges. AB - Graphene edges are of particular interest since their orientation determines the electronic properties. Here we present a detailed Raman investigation of graphene flakes with edges oriented at different crystallographic directions. We also develop a real space theory for Raman scattering to analyze the general case of disordered edges. The position, width, and intensity of G and D peaks are studied as a function of the incident light polarization. The D-band is strongest for polarization parallel to the edge and minimum for perpendicular. Raman mapping shows that the D peak is localized in proximity of the edge. For ideal edges, the D peak is zero for zigzag orientation and large for armchair, allowing in principle the use of Raman spectroscopy as a sensitive tool for edge orientation. However, for real samples, the D to G ratio does not always show a significant dependence on edge orientation. Thus, even though edges can appear macroscopically smooth and oriented at well-defined angles, they are not necessarily microscopically ordered. PMID- 19290609 TI - Enabling ultrahigh plastic flow and work hardening in twinned gold nanowires. AB - By using molecular dynamics simulations, we show that significant strain hardening and ultrahigh flow stresses are enabled in gold nanowires containing coherent (111) growth twins when balancing nanowire diameter and twin boundary spacing at the nanoscale. A fundamental transition in mechanical behavior occurs when the ratio of diameter to twin boundary spacing is larger than 2.14. A model based on site-specific dislocation nucleation and cross-slip mechanisms is proposed to explain the size dependence of flow behavior in twinned nanowires under tensile loading. PMID- 19290610 TI - Electrical conductivity of InN nanowires and the influence of the native indium oxide formed at their surface. AB - The electrical properties of InN nanowires were investigated in four-point probe measurements. The dependence of the conductance on the wire diameter allows distinguishing between "core" bulk (quadratic) and "shell" sheet (linear) contributions. Evidence of the formation of a thin In(2)O(3) layer at the surface of the nanowires is provided by X-ray core level photoemission spectroscopy. The shell conductivity is therefore ascribed to an electron accumulation layer forming at the radial InN/In(2)O(3) interface. Although conductance through the accumulation layer dominates for nanowires below a critical diameter of about 55 nm, the core channel cannot be neglected, even for small nanowires. PMID- 19290611 TI - Carbon nanotubes as injection electrodes for organic thin film transistors. AB - We have investigated the charge injection efficiency of carbon nanotube electrodes for organic semiconducting layers and compared their performance to that of traditional noble metal electrodes. Our results reveal that charge injection from a single carbon nanotube electrode is more than an order of magnitude more efficient than charge injection from metal electrodes. Moreover, organic thin film transistors that use arrays of carbon nanotube electrodes display considerable effective mobilities (0.14 cm(2)/(V.s)) and nearly ideal linear output characteristics. These results indicate that carbon nanotubes should be considered a viable alternative to metal electrodes for next-generation organic field-effect transistors. PMID- 19290612 TI - Influence of conformational flexibility on self-assembly and luminescence properties of lanthanide coordination polymers with flexible exo-bidentate biphenol derivatives. AB - To explore how nonplanar conformational distortions affect supramolecular self assembly and properties of lanthanide complexes, we have designed and synthesized two new flexible exo-bidentate ligands derived from biphenol featuring two salicylamide pendant arms, 2,2'-bis{[(2'-benzylaminoformyl)phenoxyl]ethoxyl}-1,1' biphenylene (L(I)) and 5,5'-dibromo-2,2'-bis{[(2' benzylaminoformyl)phenoxyl]ethoxyl}-1,1'-biphenylene (L(II)). These two structurally related ligands can have different conformations and are used for constructing diverse lanthanide polymers with interesting luminescence properties. Among two series of lanthanide nitrate complexes which have been characterized by elemental analysis, X-ray powder diffraction, and IR spectroscopy, four new coordination polymers have been determined using X-ray diffraction analysis. The coordination polymer type {Ln(2)(NO(3))(6)(L(I))(3).3H(2)O}(infinity) (Ln = Nd, Sm, Eu, Gd, Tb or Dy) displays a two-dimensional honeycomb-like framework in the ab plane, which can be regarded as a (6,3) topological network with neodymium atoms acting as "three connected" centers. In contrast, the coordination polymer types {[Nd(NO(3))(3)(L(II))(CH(3)OH)] x CH(3)OH}(infinity) and [Ln(NO(3))(3)(L(II))(C(2)H(5)OH)](infinity) (Ln = Sm, Eu, Gd, Tb or Dy) possess single-stranded helix chains which can be further connected through intermolecular hydrogen bonds to form two-dimensional supramolecular sheets. The photophysical properties of trivalent Sm, Eu, Tb, and Dy complexes at room temperature were investigated. The present work substantiates the claim that the supramolecular structure as well as the luminescence properties of the coordination polymer can be tuned by controlling the conformational distortion of a nonplanar flexible ligand in the supramolecular self-assembly. PMID- 19290613 TI - Using spectroscopic ellipsometry to characterize and apply the optical constants of hollow gold nanoparticles. AB - In this paper, we report the optical constants (refractive index, extinction coefficient) of self-assembled hollow gold nanoparticle (HGN) monolayers determined through spectroscopic ellipsometry (SE). We prepared a series of HGNs exhibiting various morphologies and surface plasmon resonance (SPR) properties. The extinction coefficient (k) curves of the HGN monolayers exhibited strong SPR peaks located at wavelengths that followed similar trends to those of the SPR positions of the HGNs in solution. The refractive index (n) curves exhibited an abnormal dispersion that was due to the strong SPR extinction. The values of Deltan and kmax both correlated linearly with the particle number densities. From a comparison of the optical constant values of HGNs with those of solid Au nanoparticles (NPs), we used SE measurements to demonstrate a highly sensitive Si based chemical sensor. HGNs display a slightly lower value of k at the SPR peak but a much higher sensitivity to changes in the surrounding medium than do solid Au NPs. PMID- 19290614 TI - Synthesis, characterization, and application of vanadium-salan complexes in oxygen transfer reactions. AB - We report the synthesis and characterization of several chiral salen- and salan type ligands and their vanadium complexes, which are derived from salicylaldehyde or salicylaldehyde derivatives and chiral diamines (1R,2R-diaminocyclohexane, 1S,2S-diaminocyclohexane, and 1S,2S-diphenylethylenediamine). The structures of H(2)sal(R,R-chan)(2+) x 2 Cl(-) x (CH(3))(2)CHOH x H(2)O (1c; H(2)sal(R,R-chan) = N,N'-salicyl-R,R-cyclohexanediaminium), Etvan(S,S-chen) (3c; Etvan(S,S-chen) = N,N'-3-ethoxy-salicylidene-S,S-cyclohexanediiminato), and naph(R,R-chen) (6c; naph(R,R-chen) = N,N'-naphthylidene-R,R-cyclohexanediiminato) were determined by single-crystal X-ray diffraction. The corresponding vanadium(IV) complexes and several other new complexes involving different salicylaldehyde-type precursors were prepared and characterized in the solid state and in solution by spectroscopic techniques: UV-vis, circular dichroism, electron paramagnetic resonance, and (51)V NMR, which provide information on the coordination geometry. The salan complexes oxidize in organic solvents to V(V) species, and this process was also studied using spectroscopic techniques. Single crystals suitable for X ray diffraction were obtained for [{V(V)O[sal(S,S-dpan)]}(2)(mu-O)] x H(2)O x 2(CH(3))(2)CHOH (14c; sal(S,S-dpan) = N,N'-salicyl-S,S-diphenylethylenediaminato) and [{V(V)O[t-Busal(R,R-chan)]}(2)(mu-O)] x 2 (CH(3))(2)CHOH (15c), both containing an OV(V)(mu-O)V(V)O moiety (V(2)O(3)(4+) core) with tetradentate ligands and one mu-oxo bridge. Both structures are the first examples of dinuclear vanadium complexes involving the V(V)(2)O(3)(4+) core with tetradentate ligands, the configuration of the V(2)O(3) unit being twist-angular. The V-salen and V-salan complexes are tested as catalysts in the oxidation of styrene, cyclohexene, cumene, and methyl phenyl sulfide with H(2)O(2) and t-BuOOH as oxidants. Overall, the V-salan complexes show higher activity and normally better selectivity in alkene oxidation and higher activity and enantioselectivity for sulfoxidation than their parent V-salen complexes, therefore being an advantageous alternative ligand system for oxidation catalysis. The better performance of V-salan complexes probably results from their significantly higher hydrolytic stability. Mechanisms for the alkene oxidation with these newly obtained V-salan compounds are discussed, including the use of DFT for the comparison of several alternative mechanisms for epoxidation. PMID- 19290615 TI - Acid-sensitive Pt(II) 2,6-di(pyridin-2-yl)pyrimidin-4(1H)-one complexes. AB - Three Pt(II) 2,6-di(pyridin-2-yl)pyrimidin-4(1H)-one complexes (2-4) with chloride or 4-ethynyltolyl ancillary ligands were synthesized and characterized. The photophysical properties of 2-4 were investigated in different solvents and at different acid concentrations. Their electronic absorption and emission responses at various acid concentrations were compared to those of 5. 2-4 all exhibit a broad charge-transfer band in their electronic absorption spectra from 380 to 500 nm and emit at about 560 nm in acetonitrile at room temperature, presumably ascribed to the triplet metal-to-ligand charge transfer ((3)MLCT) state. All of them exhibit broad and relatively strong triplet transient absorption in the visible to the near-IR region (450-820 nm). Upon addition of p toluenesulfonic acid, the original charge-transfer band intensity decreases, accompanied by the increase of the absorption in the 350-400 nm region in their electronic absorption spectra. Meanwhile, the (3)MLCT emission is quenched, and the triplet transient absorption intensity decreases. The changes in electronic absorption, emission, and the transient absorption spectra are reversible upon the addition of base, that is, triethylamine. The reversible acid sensitivity is caused by the protonation and deprotonation of the carbonyl oxygen on the terdentate ligand. Therefore, these platinum complexes could be potential chromogenic and luminescent sensors for acids. PMID- 19290616 TI - Studies on the atropisomerism of Fe(II) 2,6-bis(N-arylimino)pyridine complexes. AB - NMR spectra of free 2,6-bis(N-arylimino)pyridine (PDI) ligands displaying different substituents at the ortho and ortho' positions of the two N-aryl rings indicate that they can exist in syn (meso) and anti (chiral) configurations. These interconvert in solution at room temperature, via rotation of the aryl group. The corresponding paramagnetic FeX(2)(PDI) complexes exhibit the same kind of isomerism, a property that is thought to be important for their activity as alpha-olefin polymerization catalysts. For the first time, this has been detected by (1)H NMR and studied in solution. Although the conformational stability of the diastereoisomeric complexes varies widely (depending on the size of the substituents at the imine and the aromatic rings), a moderate degree of steric hindrance suffices to allow their chemical separation. A simple procedure is developed for the preparation of these complexes in diastereoisomerically pure form. In addition, introduction of a prochiral substituent in the pyridine ring enables positive assignment of the stereoisomers. Isomerization rate measurements of the Fe(II) complexes in solution suggest that isomerization very likely involves the dissociation of the corresponding Fe-N(imino) bond prior to the rotation of N-aryl groups. DFT calculations provide additional support to the conformational assignment as well as the dissociative isomerization mechanism. PMID- 19290617 TI - Is 2.07 A a record for the shortest Pt-S distance? Revision of two reported X-ray structures. AB - The comparison of the very similar compounds (Ph(3)P)(2)Pt(mu-S)(2)Pt(PPh(3))(2) (1) and (Ph(2)PyP)(2)Pt(mu-S)(2)Pt(PPh(2)Py)(2) (2) raises intriguing questions about the reliability of the reported Pt(2)S(2) core in 1, where the Pt-S bonds are the shortest ever reported. Also, the trans-annular S...S separation of 2.69 A is surprisingly shorter in 1 than in 2 (3.01 A), but no incipient coupling between two S(2-) bridges seems reasonable in this case. Various considerations lead to reformulate 1 as [(Ph(3)P)(2)Pt(mu-OH)(2)Pt(PPh(3))(2)](BF(4))(2), 3. The sets of cell parameters for 1 and 3 are not equal but two axes match, and the volume of 1 is exactly double. Simple matrices may be constructed to interconvert the direct and reciprocal crystalline cells, thus corroborating their identity of the compounds. It is concluded that, in the structure solution of 1, some atoms were either neglected (BF(4)(-) counterions) or ill identified (sulfido in place of hydroxo bridges), while the structure of 3 was solved by collecting only one half of the possible reflections (hence, also the different space groups). A new preparation, crystallization and X-ray structure of 3 confirms the above points and dismisses any other theoretical conjecture about two electronically different Pt(2)S(2) cores in 1 and 2. PMID- 19290618 TI - Comparative proteomic profiling of membrane proteins in rat cerebellum, spinal cord, and sciatic nerve. AB - Proteomics is an increasingly powerful technology that can provide in-depth insights into entire proteomes and their variation upon disease. Large-scale proteomics today enables identification and measurement of changes of thousands of proteins from minute amount of tissues. Here, we provide a proteomic profile of three distinct parts of the murine nerve system: cerebellum, spinal cord, and sciatic nerve. We focus on membrane proteins as the key regulators of neural transmission and memory. Rat tissues were homogenized and extracted to remove nonmembrane proteins and the resulting membranes were solubilized with detergents. Proteins were fractionated by size exclusion chromatography, depleted for detergents, digested and analyzed by LC-MS/MS using the LTQ-Orbitrap instrument. With the application of stringent identification criteria, in total, 4124 proteins were identified. Of these proteins, 3528, 3290, and 1649 were mapped to cerebellum, spinal cord, and sciatic nerve, respectively, allowing in depth mapping of neurotransmitter receptors, ion channels, and transporter proteins. This work is the most in-depth proteomic analysis of nerve tissues to date and provides the first unbiased insights into the proteomes of anatomically and functionally distinct parts of the membrane proteome of the central and peripheral nerve systems. The methods applied here can be directly applied to studying nerve systems and their disorders. PMID- 19290619 TI - Does the different proteomic profile found in apical and basal leaves of spinach reveal a strategy of this plant toward cadmium pollution response? AB - Chlorosis develops in Spinacia oleracea L. plants exposed to Cd and is prevalently localized in the basal leaves. A proteomic comparison of basal and apical leaves from Cd-treated plants showed modified profiles that are different and complementary in the two locations. Total chlorophyll increased in apical leaves as did photosynthetic complexes and enzymes involved in CO2 fixation and carbohydrate metabolism. Thus, apical leaves seem to supply the plant's energy requirements and, consistent with this, remain green after 40 days. In contrast, basal leaves experienced reduced chlorophyll a synthesis and photosynthesis, and later on an over production of ROS, which induces a cell defense response, leading to senescence and cell death. There was also an over production of GSH and phytochelatins, whose main role is in chelating Cd. These chelate-polypeptide complexes accumulate in the vacuole, limiting the distribution of Cd to apical leaves. In line, we found that many proteins involved in carbon metabolism were less abundant, whereas proteins involved in remobilizing carbon from other energy sources were up-regulated. We suggest that phytochelatin production has priority in Cd-stressed basal leaves and the nitrogen and sulfur metabolic pathways are activated for this purpose. Finally, as dead leaves detach from the plant, they carry away the sequestered Cd, thereby removing it completely from the plant and preventing any future access to the apical leaves. These events may represent an active detoxification strategy in higher plants. PMID- 19290620 TI - Phosphoproteome study reveals Hsp27 as a novel signaling molecule involved in GDNF-induced neurite outgrowth. AB - Glial-cell-line-derived neurotrophic factor (GDNF) is a most potent survival factor for dopaminergic neurons. In addition, GDNF was also found to promote neurite outgrowth in dopaminergic neurons. However, despite the potential clinical and physiological importance of GDNF, its mechanism of action is unclear. Therefore, we employed a state-of-the-art proteomic technique, DIGE (Difference in two-dimensional gel electrophoresis), to quantitatively compare profiles of phosphoproteins of PC12-GFRalpha1-RET cells (that stably overexpress GDNF receptor alpha1 and RET) 0.5 and 10 h after GDNF challenge with control. A total of 92 differentially expressed proteins were successfully identified by mass spectrometry. Among them, the relative levels of phosphorylated Hsp27 increased significantly both in 0.5 and 10 h GDNF-treated PC12-GFRalpha1-RET cells. Confocal microscopy and Western blot results showed that the phosphorylation of Hsp27 after GDNF treatment was accompanied by its nuclear translocation. After the mRNA of Hsp27 was interfered, neurite outgrowth of PC12 GFRalpha1-RET cells induced by GDNF was significantly blocked. Furthermore, the percentage of neurite outgrowth induced by GDNF was also reduced by the expression of dominant-negative mutants of Hsp27, in which specific serine phosphorylation residues (Ser15, Ser78 and Ser82) were substituted with alanine. Our data also revealed that p38 MAPK and ERK are the upstream regulators of Hsp27 phosphorylation. Hence, in addition to the numerous novel proteins that are potentially important in GDNF mediated differentiation of dopaminergic cells revealed by our study, our data has indicated that Hsp27 is a novel signaling molecule involved in GDNF-induced neurite outgrowth of dopaminergic neurons. PMID- 19290621 TI - A straightforward and highly efficient precipitation/on-pellet digestion procedure coupled with a long gradient nano-LC separation and Orbitrap mass spectrometry for label-free expression profiling of the swine heart mitochondrial proteome. AB - For label-free expression profiling of tissue proteomes, efficient protein extraction, thorough and quantitative sample cleanup and digestion procedures, as well as sufficient and reproducible chromatographic separation, are highly desirable but remain challenging. However, optimal methodology has remained elusive, especially for proteomes that are rich in membrane proteins, such as the mitochondria. Here, we describe a straightforward and reproducible sample preparation procedure, coupled with a highly selective and sensitive nano LC/Orbitrap analysis, which enables reliable and comprehensive expression profiling of tissue mitochondria. The mitochondrial proteome of swine heart was selected as a test system. Efficient protein extraction was accomplished using a strong buffer containing both ionic and nonionic detergents. Overnight precipitation was used for cleanup of the extract, and the sample was subjected to an optimized 2-step, on-pellet digestion approach. In the first step, the protein pellet was dissolved via a 4 h tryptic digestion under vigorous agitation, which nano-LC/LTQ/ETD showed to produce large and incompletely cleaved tryptic peptides. The mixture was then reduced, alkylated, and digested into its full complement of tryptic peptides with additional trypsin. This solvent precipitation/on-pellet digestion procedure achieved significantly higher and more reproducible peptide recovery of the mitochondrial preparation than observed using a prevalent alternative procedure for label-free expression profiling, SDS PAGE/in-gel digestion (87% vs 54%). Furthermore, uneven peptide losses were lower than observed with SDS-PAGE/in-gel digestion. The resulting peptides were sufficiently resolved by a 5 h gradient using a nano-LC configuration that features a low-void-volume, high chromatographic reproducibility, and an LTQ/Orbitrap analyzer for protein identification and quantification. The developed method was employed for label-free comparison of the mitochondrial proteomes of myocardium from healthy animals versus those with hibernating myocardium. Each experimental group consisted of a relatively large number of animals (n = 10), and samples were analyzed in random order to minimize quantitative false-positives. With this approach, 904 proteins were identified and quantified with high confidence, and those mitochondrial proteins that were altered significantly between groups were compared with the results of a parallel 2D-DIGE analysis. The sample preparation and analytical strategy developed here represents an advancement that can be adapted to analyze other tissue proteomes. PMID- 19290622 TI - Strategies for a reliable biostatistical analysis of differentially expressed spots from two-dimensional electrophoresis gels. AB - We performed quantitative comparisons with the two-dimensional gel electrophoresis technique and evaluated the reliability of biostatistical tests for the correction of "false significant" results (alpha-error) by performing repeated runs of an experiment. Results based on uncorrected p-values yielded numerous significant differences in spot intensity which could not be replicated in two additional runs. The best strategy for avoiding these "false-positive" results was strongly dependent on the type of result. In experiments yielding very marked group differences in spot intensity, calculation of the "False Discovery Rate" (FDR) by the Benjamini and Hochberg method corrected the results with sufficient reliability. In experiments yielding relatively small (p values>0.001) group differences, up to 100% of all results which were significant in two repeated runs were excluded ("false-negative") by calculation of the FDR. In such experiments, significant differences need confirmation by repeated runs. PMID- 19290623 TI - Bioinformatics and immunologic investigation on B and T cell epitopes of Cur l 3, a major allergen of Curvularia lunata. AB - The knowledge on epitopes of proteins can help in devising new therapeutic modalities for allergic disorders. In the present study, five B (P1-P5) and five T cell (P6-P10) epitopes were predicted in silico based on sequence homology model of Cur l 3, a major allergen of Curvularia lunata. Peptides (epitopes) were synthesized and assessed for biological activity by ELISA, competitive ELISA, lymphoproliferation and cytokine profiling using Curvularia allergic patients' sera. B cell peptides showed higher IgE binding by ELISA than T cell epitopes except P6. Peptides P1-P6 achieved EC(50) at 100 ng, whereas P7-P10 required 10 mug in inhibition assays. Peripheral blood mononuclear cells from Curvularia allergic patients (n = 20) showed higher lymphoproliferation for T cell epitopes than B cell epitopes except P6 confirming the properties of B and T cell prediction. The supernatant from these patients show highest interleukin-4 release on stimulation with P6 followed by B cell peptides. P4 and P6 together identified 35/37 of Curvularia positive patients by skin tests. In summary, experimental analysis confirmed in silico predicted epitopes containing important antigenic regions of Cur l 3. P6, a predicted T cell epitope, showed the presence of a cryptic B cell epitope. Peptides P4 and P6 have potential for clinical application. The approach used here is relevant and may be used to delineate epitopes of other proteins. PMID- 19290624 TI - Polymer-modified gadolinium metal-organic framework nanoparticles used as multifunctional nanomedicines for the targeted imaging and treatment of cancer. AB - Novel nanoscale theragnostic devices were successfully prepared through attachment of well defined, multifunctional polymer chains to gadolinium (Gd) metal-organic framework (MOF) nanoparticles. Copolymers of poly(N isopropylacrylamide)-co-poly(N-acryloxysuccinimide)-co-poly(fluorescein O methacrylate) were prepared via reversible addition-fragmentation chain transfer (RAFT) polymerization. The succinimide functionality was utilized as a scaffold for attachment of both a therapeutic agent, such as methotrexate, and a targeting ligand, such as H-glycine-arginine-glycine-aspartate-serine-NH(2) peptide. Employment of a trithiocarbonate RAFT agent allowed for reduction of the polymer end groups to thiolates providing a means of copolymer attachment through vacant orbitals on the Gd(3+) ions at the surface of the Gd MOF nanoparticles. These versatile, nanoscale scaffolds were shown to be biocompatible and have cancer cell targeting, bimodal imaging, and disease treatment capabilities. This unique method provided a simple yet versatile route of producing polymer-nanoparticle theragnostic materials with an unprecedented degree of flexibility in the construct, potentially allowing for tunable loading capacities and spatial loading of targeting/treatment agents, while incorporating bimodal imaging capabilities through both magnetic resonance and fluorescence microscopy. PMID- 19290625 TI - Facile synthesis of multivalent folate-block copolymer conjugates via aqueous RAFT polymerization: targeted delivery of siRNA and subsequent gene suppression. AB - Cell specific delivery of small interfering ribonucleic acid (siRNA) using well defined multivalent folate-conjugated block copolymers is reported. Primary amine functional, biocompatible, hydrophilic-block-cationic copolymers were synthesized via aqueous reversible addition-fragmentation chain transfer (RAFT) polymerization. N-(2-hydroxypropyl)methacrylamide) (HPMA), a permanently hydrophilic monomer, was copolymerized with a primary amine containing monomer, N (3-aminopropyl)methacrylamide (APMA). Poly(HPMA) confers biocompatibility, while APMA provides amine functionality, allowing conjugation of folate derivatives. HPMA-stat-APMA was chain extended with a cationic block, poly(N-[3 (dimethylamino)propyl]methacrylamide), to promote electrostatic complexation between the copolymer and the negatively charged phosphate backbone of siRNA. Notably, poly(HPMA) stabilizes the neutral complexes in aqueous solution, while APMA allows the conjugation of a targeting moiety, thus, dually circumventing problems associated with the delivery of genes via cationically charged complexes (universal transfection). Fluorescence microscopy and gene down-regulation studies indicate that these neutral complexes can be specifically delivered to cancer cells that overexpress folate receptors. PMID- 19290626 TI - Differential proteomic analysis of subfractioned human hepatocellular carcinoma tissues. AB - To discover new potential biomarkers of HCC, we used 2-DE gel separation and MALDI-TOF-MS analysis of partially enriched nuclear fractions from liver biopsies of 20 different patients. We obtained a proteomic map of subfractioned liver samples including about 200 common protein spots, among which identified components corresponded to expression products of 52 different genes. A differential analysis of proteins from tumoral and control tissues revealed a significant change in the expression level of 16 proteins associated to cytoskeletal, stress response and metabolic functions. These data may provide novel candidate biomarkers for HCC and useful insights for understanding the mechanisms of HCC pathogenesis and progression. PMID- 19290627 TI - Wet chemistry route to hydrophobic blue fluorescent nanodiamond. AB - Hydrophobic blue fluorescent nanodiamond was synthesized by covalent linking of octadecylamine to the surface of nanodiamond particles. The material is easily dispersible in hydrophobic solvents, forming a transparent colloidal solution, and can be used in those applications where stable dispersions of nanodiamond in fuels, polymers or oils are required. Bright blue fluorescence of the octadecylamine-modified nanodiamond opens up new avenues for its use as a non toxic quantum dot analogue for biomedical imaging of cellular membranes and other hydrophobic components of biological systems. Similar surface modification can be used for other carbon nanoparticles. PMID- 19290628 TI - Bovine milk caseins and transglutaminase-treated cereal prolamins are differentially recognized by IgA of celiac disease patients according to their age. AB - The prevalence of celiac disease (CD) has increased worldwide, which could be related to some dietary proteins in infant regimens and/or new food processes, affecting CD-predisposed infants and older children or adults differentially. IgA reactivity to human and bovine caseins, as well as yogurt caseins and prolamins from wheat or maize breads, microbial transglutaminase (mTG)-treated or not, was evaluated in three patient groups: G1, <2 years old; G2, approximately 3 years old; and G3 >8 years old. Human caseins were not recognized by IgA, whereas IgA reactivity of G2 and G3 was higher to bovine milk caseins. Immunoreactivity of G1 to yogurt caseins was lower and comparable to controls, with no effects due to mTG treatment. However, mTG treatment increased reactivity of G3 to wheat and maize prolamins. IgA immunoreactivity of CD patients to caseins and mTG-treated or not prolamins was age-dependent, which could reflect a differential manifestation of the effects of such proteins on the intestinal barrier. PMID- 19290629 TI - Galactose oxidase as a model for reactivity at a copper superoxide center. AB - The mononuclear copper enzyme, galactose oxidase, has been investigated under steady-state conditions via O(2)-consumption assays using 1-O-methyl-alpha-D galactopyranoside as the sugar substrate to produce an aldehyde at the C-6 position. The rate-determining step of the oxidative half-reaction was probed through the measurement of substrate and solvent deuterium and O-18 isotope effects on k(cat)/K(m)(O(2)). The reaction conforms to a ping-pong mechanism with the kinetic parameters for the reductive half, k(cat)/K(m)(S) = 8.3 x 10(3) M(-1) s(-1) at 10 degrees C and pH 7.0, comparing favorably to literature values. The oxidative half-reaction yielded a value of k(cat)/K(m)(O(2)) = 2.5 x 10(6) M(-1) s(-1). A substrate deuterium isotope effect of 32 was measured for the k(cat)/K(m)(S), while a smaller, but significant value of 1.6-1.9 was observed on k(cat)/K(m)(O(2)). O-18 isotope effects of 1.0185 with either protiated or deuterated sugar, together with the absence of any solvent isotope effect, lead to the conclusion that hydrogen atom transfer from reduced cofactor to a Cu(II) superoxo intermediate is fully rate-determining for k(cat)/K(m)(O(2)). The measured O-18 isotope effects provide corroborative evidence for the reactive superoxo species in the dopamine beta-monooxygenase/peptidylglycine alpha hydroxylating monooxygenase family, as well as providing a frame of reference for copper-superoxo reactivity. The combination of solvent and substrate deuterium isotope effects rules out solvent deuterium exchange into reduced enzyme as the origin of the relatively small substrate deuterium isotope effect on k(cat)/K(m)(O(2)). These data indicate fundamental differences in the hydrogen transfer step from the carbon of substrate vs the oxygen of reduced cofactor during the reductive and oxidative half-reactions of galactose oxidase. PMID- 19290630 TI - Wrenchnolol derivative optimized for gene activation in cells. AB - Naturally occurring transcription factors usually have two independent domains, a DNA-binding domain and an activation domain. In designing a synthetic small molecule that mimics a transcription factor, each of the two domains needs to be replaced by small-molecule counterparts. Results of the present study show that derivatives of wrenchnolol, a synthetic molecule that interacts with Sur-2 coactivator, serve as activation modules and stimulate gene transcription in vitro and in cells when tethered to a DNA-binding molecule. Thirteen derivatives of wrenchnolol were chemically synthesized and tested for their ability to activate transcription in vitro and in cells. When tethered to the GAL4 DNA binding domain, one derivative increased transcription of a GAL4-responsive reporter gene in cells 9-fold. This optimized derivative also induced up to 45% myogenesis of C2C12 cells when tethered to the DNA-binding domain of myogenic transcription factor MyoD. This optimized derivative may serve as a starting point for designing biological tools or components of fully synthetic transcription factors that permit selective up-regulation of genes. PMID- 19290631 TI - Impact of metallophilicity on "colossal" positive and negative thermal expansion in a series of isostructural dicyanometallate coordination polymers. AB - Five isostructural dicyanometallate coordination polymers containing metallophilic interactions (In[M(CN)(2)](3) (M = Ag, Au), KCd[M(CN)(2)](3), and KNi[Au(CN)(2)](3)) were synthesized and investigated by variable-temperature powder X-ray diffraction to probe their thermal expansion properties. The compounds have a trigonal unit cell and show positive thermal expansion (PTE) in the ab plane, where Kagome sheets of M atoms reside, and negative thermal expansion (NTE) along the trigonal c axis, perpendicular to these sheets. The magnitude of thermal expansion is unusually large in all cases (40 x 10(-6) K(-1) < |alpha| < 110 x 10(-6) K(-1)). The system with the weakest metallophilic interactions, In[Ag(CN)(2)](3), shows the most "colossal" thermal expansion of the series (alpha(a) = 105(2) x 10(-6) K(-1), alpha(c) = -84(2) x 10(-6) K(-1) at 295 K), while systems containing stronger Au-Au interactions show relatively reduced thermal expansion. Thus, it appears that strong metallophilic interactions hinder colossal thermal expansion behavior. Additionally, the presence of K(+) counterions also reduces the magnitude of thermal expansion. PMID- 19290632 TI - Multi-stimuli sensitive amphiphilic block copolymer assemblies. AB - Stimuli-responsive polymers are arguably the most widely considered systems for a variety of applications in biomedical arena. We report here a novel triple stimuli sensitive block copolymer assembly that responds to changes in temperature, pH and redox potential. Our block copolymer design constitutes an acid-sensitive THP-protected HEMA as the hydrophobic part and a temperature sensitive PNIPAM as the hydrophilic part with an intervening disulfide bond. The micellar properties and the release kinetics of the encapsulated guest molecule in response to one stimulus as well as combinations of stimuli have been evaluated. Responsiveness to combination of stimuli not only allows for fine tuning the guest molecule release kinetics, but also provides the possibility of achieving location-specific delivery. PMID- 19290633 TI - Reductive half-reaction of aldehyde oxidoreductase toward acetaldehyde: a combined QM/MM study. AB - We report a combined QM/MM study on the mechanism of the reductive half-reaction of aldehyde oxidoreductase. Five possible pathways are explored concerning the binding mode of acetaldehyde and the catalytic effect of the nearby glutamic acid (Glu869), taking both possible protonation states into account. In the most favorable pathway, Glu869 participates and acts as a Lewis base to deprotonate the labile hydroxide group. This proton transfer is essential for the high activity of the enzyme toward substrate because it increases the nucleophilicity of the migrating O atom and strengthens the electrophilicity of the target C atom in the substrate. The subsequent product-forming reactions occur in two discrete steps, first nucleophilic attack and then hydride transfer, which implies that the oxidation of aldehyde is a two-electron process. A variant of this mechanism, with an additional water molecule bridging the Glu869 side chain and the substrate, has similar barriers. Judging from previous gas phase calculations and our present QM/MM data, the catalytic effect of Glu869 mainly lowers the barrier of the nucleophilic attack so that the hydride transfer becomes the rate determining step in the reductive half-reaction. PMID- 19290634 TI - Quasi-planar homopolymetallic and heteropolymetallic coordination arrays. Surface like molecular clusters of magnesium and aluminum. AB - The sterically isolated preorganized tetradentate ligand systems, tetrakis(2 hydroxy-3-n-propylphenyl)ethene and tetrakis(5-tert-butyl-2-hydroxy-3 trimethylsilylphenyl)ethene, nucleate the formation of quasi-planar raft-like polymetallic coordination complexes with high selectivity, providing topologically consistent structural models for metal coordination to the "oxo surface" of silica- and alumina-supported heterogeneous catalysts. The coordination of magnesium salts to these systems yields trimetallic magnesium halide and alkyl complexes arrayed on the oxygen "surface" of the ligand, regardless of the steric profile of the ortho-substituents. The magnesium complexes, characterized in the solid state by X-ray crystallography, contain two chemically distinct metal environments, a relatively inert central magnesium bis(alkoxide) and two more labile pseudotetrahedral "wing" magnesium atoms. The central metal coordination is pseudo-octahedral; crystallography strongly suggests the presence of an unprecedented dative magnesium-olefin bonding interaction from the metal to ethene bridge of the ligand. Consistent with the chemistry proposed for typical magnesium-treated catalyst supports, the labile wing magnesium centers can be cleanly and sequentially exchanged for aluminum with retention of the surface-like coordination array. Thus, treatment with diethylaluminum chloride provides heterotrimetallic magnesium-aluminum complexes containing one aluminum and two magnesium sites or two aluminum and one magnesium site, respectively. All four heteropolymetallic complexes have been characterized by X-ray crystallography. PMID- 19290635 TI - Quantitative three-dimensional modeling of zeotile through discrete electron tomography. AB - Discrete electron tomography is a new approach for three-dimensional reconstruction of nanoscale objects. The technique exploits prior knowledge of the object to be reconstructed, which results in an improvement of the quality of the reconstructions. Through the combination of conventional transmission electron microscopy and discrete electron tomography with a model-based approach, quantitative structure determination becomes possible. In the present work, this approach is used to unravel the building scheme of Zeotile-4, a silica material with two levels of structural order. The layer sequence of slab-shaped building units could be identified. Successive layers were found to be related by a rotation of 120 degrees, resulting in a hexagonal space group. The Zeotile-4 material is a demonstration of the concept of successive structuring of silica at two levels. At the first level, the colloid chemical properties of Silicalite-1 precursors are exploited to create building units with a slablike geometry. At the second level, the slablike units are tiled using a triblock copolymer to serve as a mesoscale structuring agent. PMID- 19290636 TI - Highly porous and robust 4,8-connected metal-organic frameworks for hydrogen storage. AB - Highly porous and robust metal-organic frameworks (MOFs) were constructed based on aromatics-rich octa-carboxylate ligands and copper paddle-wheel building units. Each octa-carboxylate ligand is linked to eight copper paddle wheels via the bridging carboxylate groups in a rectangular prismatic fashion to lead to very rare (4,8)-connected networks of the scu topology. The high-connectivity MOFs show remarkably high porosity and framework stability, as evidenced by a perfect agreement between experimental and theoretical surface areas and the maintenance of framework powder X-ray diffraction patterns after solvent removal. These aromatics-rich MOFs exhibit an exceptionally high hydrogen uptake of up to 2.5 wt% at 77 K and 1 atm. This work thus demonstrates the ability to construct highly porous and robust functional MOFs using multidentate bridging ligands of high connectivity. Such a rational synthetic strategy is complementary to the common reliance on high-nuclearity metal clusters for building stable and porous MOFs. PMID- 19290637 TI - Evaluation of two food grade proliposomes to encapsulate an extract of a commercial enzyme preparation by microfluidization. AB - The entrapment by microfluidization of a commercial enzyme extract (Debitrase DBP20) in liposomes using two food grade proliposome (C and S) preparations was studied. Liposomes obtained at a low microfluidization pressure (4000 psi) were distributed in a bimodal population of small (30-40 nm) and large vesicles (300 700 nm). The composition of the proliposome influenced entrapment efficiency and the repartition of the enzyme between the core and the surface of the liposome. More enzyme was associated with the liposomal surface and greater entrapment efficiencies (64%) were obtained for liposomes with the highest negative zeta potential (proliposome C). Increasing microfluidization pressure and increasing the number of passes through the microfluidizer resulted in losses in entrapment efficiency and enzyme activity, due to decreasing liposome size and enzyme denaturation. Entrapment efficiency was not influenced by external pH and enzyme activity was not adversely affected over storage for 18 days under the conditions evaluated. PMID- 19290638 TI - Galactooligosaccharides (GOS) inhibit Vibrio cholerae toxin binding to its GM1 receptor. AB - It is widely reported that cholera toxin (Ctx) remains a significant cause of gastrointestinal disease globally, particularly in developing countries where access to clean drinking water is at a premium. Vaccines are prohibitively expensive and have shown only short-term protection. Consequently, there is scope for continued development of novel treatment strategies. One example is the use of galactooligosaccharides (GOS) as functional mimics for the cell-surface toxin receptor (GM1). In this study, GOS fractions were fractionated using cation exchange chromatography followed by structural characterization using a combination of hydrophilic interaction liquid chromatography (HILIC) and electrospray ionization mass spectrometry (ESI-MS) such that their molecular weight profiles were known. Each profile was correlated against biological activity measured using a competitive inhibitory GM1-linked ELISA. GOS fractions containing >5% hexasaccharides (DP(6)) exhibited >90% binding, with EC(50) values between 29.27 and 56.04 mg/mL. Inhibition by GOS DP(6) was dose dependent, with an EC(50) value of 5.10 mg/mL (5.15 microM MW of 990 Da). In removing low molecular weight carbohydrates that do possess prebiotic, nutraceutical, and/or biological properties and concentrating GOS DP(5) and/or DP(6), Ctx antiadhesive activity per unit of (dry) weight was improved. This could be advantageous in the manufacture of pharmaceutical or nutraceutical formulations for the treatment or prevention of an acute or chronic disease associated with or caused by the adhesion and/or uptake of a Ctx or HLT. PMID- 19290639 TI - Nutriomic analysis of fresh and processed fruit products. 1. During in vitro digestions. AB - Nutriomic analysis is a postgenomic-based study of nutritious components (nutriome). There is a need for an in vitro digestion and absorption model to unravel interactive factors varying nutriome release from various food materials that cannot be directly studied in humans. Effects of processing and in vitro digestion steps on carotenoid, sugar, and organic acid release from tomato, papaya, and mango products were comprehensively studied for the first time in this research. In vivo chewing experiments using 24 healthy adult volunteers was carried out prior to chewing simulation. Microscopy showed that cutting and blending alone were unlikely to mimic chewing at swallowing point. Using general linear model (GLM) ANOVA and principal component analysis (PCA), effects of interaction between digestion steps and processing types on the nutriome release were significant (p < 0.05) when 90% particles of 0.5 (dried) and 1.5 cm (fresh) were digested in vitro. Generally, dried and fresh fruits released lower levels of nutriome components than juices. PCA indicated nutriome release from tomato products was affected by the factors studied more than those from papaya and mango products. Fruit type is the main determinant factor relative to processing and digestion steps because it determines the extent of matrix that breaks down and consequent nutriome diffusion rates. It is predicted that pectin plays a role in determining the rate of nutriome release and absorption, which requires further investigation. PMID- 19290640 TI - Nutriomic analysis of fresh and processed fruit products. 2. During in vitro simultaneous molecular passages using Caco-2 cell monolayers. AB - Many studies have used Caco-2 cell monolayers as human intestinal absorption models. However, only a few studied digested foods, instead of pure standard compounds. Moreover, beneficial and nutritional molecules (nutriome) have not been investigated simultaneously. The present study explored nutriome passages from digest solution of fresh, dried, and juiced tomato, mango, and papaya using Caco-2 cell monolayers in apical-->basolateral directions. A validation method using complementary TEER and P(app) values or internal standard caffeine is recommended because physiologically passive diffusion is unlikely to happen. Sugars were transported into basolateral sides, resulting in potential glucose equivalent bioavailability of 2.26-75 mg h(-1)/100 g (WB). Using sugar passage rates (DB) of juices as 100% references, the rate order was tomato (49.8% dried; 89.5% fresh) > mango (56.8% dried; 22.8% fresh) > papaya (18.7% dried; 36.7% fresh). Major indications that phytochemical absorption does not occur in the small intestine were obtained from the bioassay condition selected. Apical organic acid levels decreased, which occasionally were transported into basolateral sides, whereas the disappearances of apical carotenoids and phenolics were not. Pectin substances were predicted to be responsible for the disappearances of bioactive compounds in those pectin-rich fruits. Further investigations on the role of pectin substances in intestinal passages are recommended. PMID- 19290641 TI - Oxidative stability of unsaturated monoacyl trehalose in aqueous solution. PMID- 19290642 TI - Novel approach for chemotype hopping based on annotated databases of chemically feasible fragments and a prospective case study: new melanin concentrating hormone antagonists. AB - A novel strategy for chemotype hopping, based on annotated databases of chemically feasible fragments and their oriented functionalization, is presented. A three-dimensional (3D) similarity analysis of project-oriented functionalized scaffolds provides a prioritized proposal for synthesis with the most appropriate linkers and optimal regiochemistry on R-groups. This strategy maximizes the potential of proprietary and commercially available compounds. A retrospective and prospective case study, on melanin concentrating hormone (MCH) antagonists, showing the impact on the drug discovery process of this new strategy by maintaining primary activity and improving key ADME/Tox property while enhancing intellectual property (IP) position is demonstrated. PMID- 19290643 TI - Nanoimprinted thin films of reactive, azlactone-containing polymers: combining methods for the topographic patterning of cell substrates with opportunities for facile post-fabrication chemical functionalization. AB - Approaches to the fabrication of surfaces that combine methods for the topographic patterning of soft materials with opportunities for facile, post fabrication chemical functionalization could contribute significantly to advances in biotechnology and a broad range of other areas. Here, we report methods that can be used to introduce well-defined nano- and microscale topographic features to thin films of reactive polymers containing azlactone functionality using nanoimprint lithography (NIL). We demonstrate that NIL can be used to imprint topographic patterns into thin films of poly(2-vinyl-4,4-dimethylazlactone) and a copolymer of methyl methacrylate and 2-vinyl-4,4-dimethylazlactone using silicon masters having patterns of grooves and ridges ranging in width from 400 nm to 2 microm, demonstrating the potential of this method to transfer patterns to films of these reactive polymers over a range of feature sizes and densities. We demonstrate further that the azlactone functionality of these polymers survives temperatures and pressures associated with NIL, and that topographically patterned films can be readily functionalized post-fabrication by treatment of surface-accessible azlactone functionality with small molecules and polymers containing primary amines. The results of experiments in which NIH-3T3 cells were seeded onto films imprinted with lined patterns having a pitch of 4 microm demonstrated that cells attach and proliferate on these azlactone-containing films and that they align in the direction of the imprinted pattern. Finally, we demonstrate that the treatment of these materials with amine-functionalized poly(ethylene glycol) (PEG) can be used to create regions of topographically patterned films that prevent cell adhesion. The results of this study suggest approaches to the functionalization of topographically patterned surfaces with a broad range of chemical functionality (e.g., peptides, proteins, carbohydrates, etc.) of biotechnological interest. The ability to manipulate and define both the physical topography and chemical functionality of these reactive materials could provide opportunities to investigate the combined effects of substrate topography and chemical functionality on cell behavior and may also be useful in a broad range of other applications. PMID- 19290644 TI - Near-field enhancement of multipole plasmon resonances in Ag and Au nanowires. AB - In this paper, we investigate theoretically the electromagnetic field enhancement arising from excitation of silver and gold nanowires (NWs) of finite length, capable of sustaining surface plasmon resonances of different multipole order, using the Discrete Dipole Approximation (DDA). The influence of NW length on the degree of enhancement and confinement of the electromagnetic field for each surface plasmon mode is analyzed by a 3D mapping of the near field for different planes around the NW as well by calculating its variation with distance along two different directions, one parallel to and the other perpendicular to the NW axis, outside of the NW. It was found that the enhancement is still significant at relative large distances from the NW end, its decay being of much longer range than that predicted by a simple dipole approximation, especially at near-infrared wavelengths. PMID- 19290645 TI - Spontaneous formation of dipolar metal nanoclusters. AB - The adsorption of three- and four-atom Ag and Pd clusters on the alpha-Al(2)O(3) (0001) surface is explored with density functional theory. Within each adsorbed cluster, two different cluster-surface interactions are present. We find that clusters simultaneously form both ionic bonds with surface oxygen and intermetallic bonds with surface aluminum. The simultaneous formation of disparate electronic structure motifs within a single metal nanoparticle is termed a "dipolar nanocluster". This coexistence is ascribed to a balance of geometric constraints and metal electronic structure, and its importance for nanoparticle catalysis is highlighted. PMID- 19290646 TI - Folding and association of thermophilic dimeric and trimeric DsrEFH proteins: Tm0979 and Mth1491. AB - Although the majority of natural proteins exist as protein-protein complexes, the molecular basis for the formation and regulation of such interactions and the evolution of protein interfaces remain poorly understood. We have investigated these phenomena by characterizing the thermal and chemical denaturation of thermophilic DsrEFH proteins that have a common subunit fold but distinct quaternary structures: homodimeric Tm0979 and homotrimeric Mth1491. Tm0979 forms a moderate affinity dimer, and a monomeric intermediate is readily populated at equilibrium and during folding kinetics. In contrast, the Mth1491 trimer has extremely high stability, so that a monomeric form is not measurably populated at equilibrium, although it may be during folding kinetics. A common mechanism for evolution of quaternary structures may be facile formation of a relatively stable monomeric species, with stabilizing intermolecular interactions centering on alternative environments for a beta-strand at the edge of the monomer, augmented by malleable hydrophobic interactions. The exceptional trimer stability arises from a remarkably slow unfolding rate constant, 6.5 x 10(-13) s(-1), which is a common characteristic of highly stable thermophilic and/or oligomeric proteins. The folding characteristics of Tm0979 and Mth1491 have interesting implications for assembly and regulation of homo- and heterooligomeric proteins in vivo. PMID- 19290647 TI - Dendritic effect on supramolecular self-assembly: organogels with strong fluorescence emission induced by aggregation. AB - A novel class of dumbbell-shaped dendritic molecules with a p-terphenylene core was synthesized, and their self-assembling properties were investigated. The incorporation of bulky dendritic wedges to the central stiff aromatic scaffolds could finely tune their solubility in many organic solvents. Unlike the self assembly behavior of p-terphenylen-1,4"-ylenebis(dodecanamide), the p terphenylene cored different generation dendritic molecules could form gels in several kinds of organic solvents through a cooperative effect of the pi-pi stacking, hydrogen-bonding, and van der Waals forces. Interestingly, significant fluorescence enhancement was observed after gelation. Extensive investigations with atomic force microscopy (AFM), scanning electron microscopy (SEM), transmission electron microscopy (TEM), theological measurements, UV-vis absorption spectroscopy, FT-IR spectroscopy, 1H NMR, and X-ray powder diffraction (XRD) revealed that these dendritic molecules self-assembled into elastically interpenetrating one-dimensional nanostructures in organogels. PMID- 19290648 TI - Triterpenoid saponins from Dianthus versicolor. AB - Seven new pentacyclic triterpenoid saponins, named dianversicosides A-G (1-7), together with nine known compounds, were isolated from the aerial parts of Dianthus versicolor. The structures of 1-7 were elucidated on the basis of spectroscopic data and chemical evidence. The absolute configuration of the 3 hydroxyl-3-methylglutaryl (HMG) group in 1-4 was ascertained by chemical analysis combined with a chiral HPLC method. Cytotoxic activities of the isolated compounds were evaluated against a small panel of cancer and other cell lines. PMID- 19290649 TI - Preparation of porous poly(L-lactic acid) honeycomb monolith structure by phase separation and unidirectional freezing. AB - A honeycomb monolith structure with micro/nanoscale porous walls was successfully fabricated in poly(l-lactic acid) (PLLA) by integrating polymer-solvent and polymer-polymer phase separations induced during a pseudosteady-state unidirectional freezing process. Poly(ethylene glycol) (PEG) and PLLA were dissolved in 1,4-dioxane to prepare a single phase polymer solution. The direction of freezing created a honeycomb monolith structure of PLLA/PEG polymers. Crystallization of the solvent reduced the solvent concentration and induced liquid-liquid phase separation during the unidirectional freezing. A sea and-island morphology, where PEG domains were dispersed in the PLLA matrix, was developed, and pores were created in the channel walls of the honeycomb monolith structure by leaching out the PEG domain. The effects of the PEG molecular weight and the PLLA/PEG weight ratio on the aligned honeycomb structure and the pores in the channel walls were investigated. A ternary phase diagram for PLLA, PEG, and 1,4-dioxane was created from cloud point temperature measurements. Based on this phase diagram, hypotheses for the mechanism of the cellular-dendritic transition and the formation mechanism of the pores in the channel walls are proposed. PMID- 19290650 TI - Natural and engineered nano and colloidal transport: role of zeta potential in prediction of particle deposition. AB - This study is designed to answer the question of whether the zeta-potential measurement of natural and engineered nano and colloidal particles can be used as a general predictor for their transport and deposition within porous medium. The transport and deposition of engineered zeolite particles and natural colloids (soil and sediment colloids) was studied and evaluated considering their zeta potential measurement. The zeta-potential of Ca(2+) or K(+) saturated zeolite particles increased (i.e., became less negative) with increasing bulk ionic strength, independent of the cation species in the bulk solution, while the zeta potential of Ca(2+) or K(+) saturated natural colloids was dependent on the cation species in the bulk solution, unexpectedly decreasing with the bulk K(+) concentrations while increasing with bulk Ca(2+) concentrations (10(-5) to 10(-2) M). The particle deposition rate coefficient (k) was sensitive to the cation species in the bulk solution and on the cation exchangeable sites of the particle surfaces. The trend of the zeta-potential of the natural colloids, especially with simple electrolytes in bulk solution, was at odds with that of k under the same conditions, indicating that the zeta-potential of these natural colloids could not be used to adequately predict their transport and deposition. The observed anomaly is a direct result of surface heterogeneity and hydrophobicity of natural colloids, and it points out the important difference between natural and engineered nano and colloidal particles in terms of their transport and deposition prediction. PMID- 19290651 TI - Magnetic-field-induced formation of one-dimensional magnetite nanochains. AB - Magnetite nanoclusters with an average size of about 120 nm have been prepared and allowed to self-assemble into one-dimensional (1D) nanochain structures with the average length about of 2 mum by a simple magnetic-field-induced (MFI) assembly approach (0.20 T). The constituent, phase, and morphology of these 1D nanochains have been characterized by X-ray diffraction and transmission electron microscopy. Magnetic measurement reveals that these 1D nanochains are weakly ferromagnetic at room temperature. In this paper, we discuss the influence of magnetization time and strength of external magnetic field on the formation of 1D nanochains. We also show that, by changing the amount of hydrogen peroxide in the starting materials, 1D nanochains with different interparticle spacing can be obtained. This 1D nanochain structure with different interparticle spacing would be an ideal system for the further study of magnetization properties of 1D ordered magnetic nanostructures. PMID- 19290652 TI - Association behavior of star-shaped pH-responsive block copolymer: four-arm poly(ethylene oxide)-b-poly(methacrylic acid) in aqueous medium. AB - A four arm pH-responsive poly(ethylene oxide)-b-poly(methacrylic acid) block copolymer was synthesized by atom transfer radical polymerization technique. The conformation transition over the course of neutralization was investigated using a combination of potentiometric and conductometric titrations, dynamic and static light scattering, and transmission electron microscopy. The multiarm block copolymer existed as an extended unimer at high pH due to the negatively charged carboxylate groups and hydrophilic poly(ethylene oxide) segments. The block copolymers self-assembled into core-shell micelles and large spherical aggregates that flocculated at low degree of neutralization (alpha). Such behavior is controlled by the fine balance of electrostatic, hydrophobic, and hydrogen bond interactions. The hydrodynamic radius (R(h)) of the aggregates was approximately 84 nm at alpha of 0.3, and it decreased to 63 and 46 nm at alpha approximately 0.2 and 0.1, respectively, as a result of the reduced electrostatic interaction between ionized carboxylate groups. The thermodynamic parameters obtained from isothermal titration calorimetric technique in different salt concentrations indicated that the energy to extract a proton from a charged polyion was reduced by the addition of salt, which favors the neutralization process. PMID- 19290654 TI - Enantio- and diastereoselective tandem zn-promoted brook rearrangement/ene-allene carbocyclization reaction. AB - The zinc-catalyzed addition of various alkynes to acylsilanes followed by a Zn Brook rearrangement and either the Zn-ene-allene or Zn-yne-allene cyclization led to the enantio- and diastereoselective formation of carbocycles in a single-pot operation. PMID- 19290655 TI - Role for threonine 201 in the catalytic cycle of the soluble diiron hydroxylase toluene 4-monooxygenase. AB - The active site residue Thr-201 in toluene 4-monooxygenase hydroxylase (T4moH) has a structural counterpart in the active sites of all diiron monooxygenases. Thus, our previous finding that mutation of this residue to Ala, Gly, or Ser had no impact on steady-state catalysis or coupling was surprising. In this work, we provide kinetic, biochemical, and structural evidence that one role of Thr-201 may be to stabilize a peroxo-level intermediate during enzyme catalysis. During reactions in the absence of substrate, T201 T4moH slowly consumed O(2) but only a negligible amount of H(2)O(2) was released. In contrast, T201A T4moH gave stoichometric release of H(2)O(2) during reaction in the absence of substrate. Both enzyme isoforms were tightly coupled during steady-state catalysis with saturating toluene and other optimal substrates and exhibited near-identical kinetic parameters. However, rapid mix single-turnover studies showed that T201A T4moH had a faster first-order rate constant for product formation than T201 T4moH did. Comparison of X-ray crystal structures of resting and reduced T201A T4moH in complex with T4moD with comparable structures of T201 T4moHD revealed changes in the positions of several key active site residues relative to the comparable structures of T201 T4moH with T4moD. This combination of catalytic and structural studies offers important new insight into the role of the role of conserved Thr-201, and its contributions to the catalytic reaction cycle. PMID- 19290656 TI - Capillary spreading dynamics of electrowetted sessile droplets in air. AB - We report the contact line dynamics of sessile water droplets, 1.1-1.6 mm in radius, spread by electrowetting in air. Coplanar electrodes patterned on the substrate allow a true sessile condition with no wire into the droplet. The frequency response of the droplets is studied using 25 VAC ranging from 10 to 205 Hz. The effect of contact angle hysteresis is seen in form of stick-slip motion. A model developed provides a good match to the experimental result. Step response is studied with voltages in the range of 20-80 VDC. Two regimes of motion are observed. In the first regime, local flows cause the contact line speed to increase and reach a maximum while the contact angle is still changing. Global flows in the second regime cause the contact line to move with a reduced speed and attain the spherical shape pertaining to the new equilibrium contact angle. A model is used to describe the motion. PMID- 19290657 TI - pH-responsive and thermoreversible hydrogels of N-(2-hydroxyalkyl)-L-valine amphiphiles. AB - Gelation behavior of a series of low-molecular-weight-hydrogelators, N-(2 hydroxydodecyl)-L-amino acid, was studied in aqueous phosphate buffer (pH 12). The effect of head-group structure and chirality and hydrocarbon chain length on the gelation efficiency was investigated. Only N-(2-hydroxyalkyl)-L-valine (L C(n)HVal, n = 10, 12, 14, and 16) derivatives were found to form gel in aqueous buffer at pH 12. The increase of the chain length of the hydrocarbon tail enhances the ability to gelate buffered water up to C14 chain length. The L C16HVal amphiphile was found to have gelation ability lower than L-C14HVal. The gelation number, mechanical strength, thermal stability, and morphology of the supramolecular aggregates were studied. The effect of salt concentrations on the gelation was investigated. Addition of NaCl increased gelation number but decreased melting temperature of the hydrogels slightly. The morphology of the hydrogels was characterized by electron microscopy and small-angle X-ray diffraction techniques. Rheology measurements were performed to examine the mechanical strength of the hydrogels. Both hydrogen-bonding and hydrophobic interactions were shown to be the driving forces for supramolecular aggregate formation. PMID- 19290658 TI - Evidence for a Ca(2+)-specific conformational change in avian thymic hormone, a high-affinity beta-parvalbumin. AB - Named for the capacity to stimulate differentiation and maturation of T-cell precursors, avian thymic hormone (ATH) is nonetheless a beta-parvalbumin that is also expressed in the avian retina. With Ca(2+)- and Mg(2+)-binding constants in excess of 10(8) and 10(4) M(-1), respectively, both EF-hand motifs qualify as Ca(2+)/Mg(2+) sites. However, whereas addition of either apo- or Mg(2+)-bound ATH to 1,8-anilinonaphthalenesulfonic acid (ANS) causes a large increase in quantum yield and a pronounced blue shift, addition of the Ca(2+)-bound protein is without effect. These observations suggest that apo- and Mg(2+)-bound ATH adopt conformations distinct from the Ca(2+)-bound protein, exposing apolar surface for interaction with ANS. Differential scanning calorimetry (DSC) data imply that unfolding of apo-ATH is accompanied by diminished exposure of apolar surface, relative to Ca(2+)-free rat beta-PV, perhaps due to greater solvent-accessible apolar surface in the native form. The fluorescence and DSC results, considered together, may indicate that the AB and CD-EF domains of ATH are not tightly associated in the absence of bound Ca(2+). Consistent with this idea, sedimentation velocity data reveal that the apo- and Mg(2+)-bound forms of ATH show greater departures from spherical symmetry than the Ca(2+)-bound state. These findings suggest that a high-affinity binding signature does not require that the parvalbumin apo- and Ca(2+)-bound conformations be indistinguishable, as we have recently proposed. They also suggest that it is possible to engineer a Ca(2+)-dependent conformational change into a high-affinity EF-hand protein, furnishing a mechanism by which the protein could play a reverse Ca(2+) sensor role. PMID- 19290659 TI - A hydrogen-substituted osmium stannylene complex: isomerization to a metallostannylene complex via an unusual alpha-hydrogen migration from tin to osmium. AB - An osmium complex bearing a terminal hydrogen-substituted stannylene ligand, Cp*((i)Pr(3)P)(H)Os=SnH(trip) (1) (trip = 2,4,6-triisopropylphenyl), has been prepared by stannylene extrusion, and the complex has been structurally characterized. Complex 1 coordinates Lewis bases and activates the O-H bonds of water and methanol. Most interestingly, 1 converts to the metallostannylene complex Cp*((i)Pr(3)P)(H)(2)OsSn(trip) (2) thermally or photochemically by what appears to be a radical process. PMID- 19290660 TI - Hydrogen- and oxygen-driven interconversion between imido-bridged dirhodium(III) and amido-bridged dirhodium(II) complexes. AB - The reaction of [Cp*RhCl(2)](2) (Cp* = eta(5)-C(5)(CH(3))(5)) with 2 equiv of p toluenesulfonamide in the presence of KOH resulted in the formation of the sulfonylimido-bridged dirhodium(III) complex [(Cp*Rh)(2)(mu-NTs)(2)] (1a; Ts = SO(2)C(6)H(4)CH(3)-p). The imido complex 1a reacted with hydrogen donors such as H(2) and 2-propanol to give the sulfonylamido-bridged dirhodium(II) complex [(Cp*Rh)(2)(mu-NHTs)(2)] (2). Treatment of the (amido)rhodium(II) complex 2 with O(2) regenerated the (imido)rhodium(III) complex 1a. Complex 1a also underwent reversible protonation to afford the cationic amido- and imido-bridged dirhodium(III) complex [(Cp*Rh)(2)(mu-NHTs)(mu-NTs)](+) (4), which further reacted with H(2) or 2-propanol to give the (hydrido)bis(amido)dirhodium(III) complex [(Cp*Rh)(2)(mu-H)(mu-NHTs)(2)](+) (5). On the basis of DFT calculations and experimental results using 4 and 5, the reaction of 1a with H(2) proved to proceed via heterolytic cleavage of H(2) assisted by the sulfonyl oxygen atom followed by proton migration from the metal center. Furthermore, the redox interconversion between 1a and 2 was applied to catalytic aerobic oxidation of H(2) and an alcohol by using 1a as a well-defined dinuclear catalyst. The iridium complex [(Cp*Ir)(2)(mu-NTs)(2)] (1b) as well as a rhodium complex [Cp*RhCl(2)](2) without bridging imido ligands did not catalyze these aerobic oxidation reactions. PMID- 19290661 TI - Overhauser dynamic nuclear polarization to study local water dynamics. AB - Surface and internal water dynamics of molecules and soft matter are of great relevance to their structure and function, yet the experimental determination under ambient and steady-state conditions is challenging. One of the most powerful approaches to measure local water dynamics within 5 A distances is to utilize the modulation of the nuclear spin relaxation rate of water protons through their time-dependent dipolar coupling to paramagnetic probes, here nitroxide spin labels. We recently introduced a method to obtain local water dynamics through Overhauser dynamic nuclear polarization (DNP). This has a unique advantage over other related techniques available in that a highly amplified proton nuclear magnetic resonance signal carries the information, allowing the use of minute microliter sample volumes and 100 muM sample concentrations. The outcome of our approach is the quantitative determination of the key DNP parameter known as the coupling factor, which provides local translational diffusion dynamics of the solvent within 5 A of the spin label. In contrast to recent reports that the coupling factor for nitroxide radicals cannot be quantified due to the difficulty in determining the saturation factor for the spin label, we show the saturation factor can be accurately determined and for the first time present agreement between measurements and theory. We discuss the discrepancy between the related field cycling relaxometery technique and DNP in determining the coupling factor and present arguments in support of the DNP determined value. DNP measurements of local hydration dynamics around nitroxides in bulk water and on the surface of proteins are presented. PMID- 19290662 TI - Chiral Bronsted acid-catalyzed enantioselective alpha-hydroxylation of beta dicarbonyl compounds. AB - A novel, facile, and highly enantioselective Bronsted acid-catalyzed alpha hydroxylation of beta-dicarbonyl compounds with up to 99:1 er using nitroso compounds as the oxygen source has been developed. The results disclosed herein considerably extend the substrate scope for the alpha-aminoxylation, allowing expeditious, straightforward, and efficient access to valuable alpha-hydroxy-beta dicarbonyl compounds with the highest levels of enantiocontrol. PMID- 19290663 TI - On the Pd-catalyzed vinylation of aryl halides with tris(alkoxy)vinylsilanes in water. AB - Pd-catalyzed vinylation of aryl halides with tris(alkoxy)vinylsilanes occurs in aqueous NaOH solution through Heck coupling of the aryl halide with the silyl olefin followed by desilylation at the Pd center or, at high temperatures, hydrolysis of the C-Si bond. Pd-mediated desilylation does not occur via beta-Si elimination under these conditions. PMID- 19290664 TI - Invariance of exocytotic events detected by amperometry as a function of the carbon fiber microelectrode diameter. AB - Etched carbon fiber microelectrodes of different radii have been used for amperometric measurements of single exocytotic events occurring at adrenal chromaffin cells. Frequency, kinetic, and quantitative information on exocytosis provided by amperometric spikes were analyzed as a function of the surface area of the microelectrodes. Interestingly, the percentage of spikes with foot (as well as their own characteristics), a category revealing the existence of sufficient long-lasting fusion pores, was found to be constant whatever the microelectrode diameter was, whereas the probability of overlapping spikes decreased with the electrode size. This confirmed that the prespike foot could not feature accidental superimposition of separated events occurring at different places. Moreover, the features of amperometric spikes investigated here (charge, intensity and kinetics) were found constant for all microelectrode diameters. This demonstrated that the electrochemical measurement does not introduce significant bias onto the kinetics and thermodynamics of release during individual exocytotic events. All in all, this work evidences that information on exocytosis amperometrically recorded with the usual 7 microm diameter carbon fiber electrodes is biologically relevant, although the frequent overlap between spikes requires a censorship of the data during the analytical treatment. PMID- 19290665 TI - Imaging and quantification of isotachophoresis zones using nonfocusing fluorescent tracers. AB - We present a novel method for visualizing isotachophoresis (ITP) zones. We introduce negligibly small concentrations of a fluorophore that is not focused by isotachophoresis. This nonfocusing tracer (NFT) migrates through multiple isotachophoresis zones. As it enters each zone, the NFT concentration adapts to the local electric field in each zone. ITP zones can then be visualized with a point detector or camera. The method can be used to detect, identify, and quantify unknown analyte zones and can visualize complex and even transient electrophoresis processes. This visualization technique is particularly suited to microfluidic and laboratory-on-a-chip applications, as typical fluorescence microscopes and charge-coupled device (CCD) cameras can provide high-resolution spatiotemporal data. We present a theoretical description, a methodology for identifying analytes, and experimental validation. We also visualize and analyze a complex, transient DNA ITP preconcentration and separation. PMID- 19290666 TI - Direct profiling and imaging of epicuticular waxes on Arabidopsis thaliana by laser desorption/ionization mass spectrometry using silver colloid as a matrix. AB - Colloidal silver laser desorption/ionization (LDI) mass spectrometry (MS) was employed to directly profile and image epicuticular wax metabolites on a variety of different surfaces of Arabidopsis thaliana leaves and flowers. Major cuticular wax compounds, such as very long-chain fatty acids, alcohols, alkanes, and ketones, were successfully detected as silver adduct ions. The surface metabolites of different flower organs (carpels, petals, and sepals) were profiled for the first time at a spatial resolution of approximately 100 microm. In addition, mass spectral profiles and images were collected from wild type and a mutant strain, which carried alleles that affect the surface constituents of this organism. One of these mutant alleles (cer2-2) is in a gene whose biochemical functionality is still unclear, although its effect on normal epicuticular wax deposition was the characteristic that led to its original identification. Variations of wax products between different spatial locations for wild type and for a mutant strain were investigated by normalizing the ion intensities to a reference peak ([(107)Ag + (109)Ag](+)). The spatially resolved surface metabolite profiling data of this mutant has provided new insights into the complexity of epicuticular wax deposition at the cellular-resolution scale. This MS-based metabolite imaging technology has the potential to provide valuable data for dissecting metabolism in multicellular organism at the level of single cells. PMID- 19290667 TI - Detection of extracellular H2O2 released from human liver cancer cells based on TiO2 nanoneedles with enhanced electron transfer of cytochrome c. AB - The high conductive TiO(2) nanoneedles film is first employed as a support matrix for immobilizing model enzyme, cytochrome c (cyt c) to facilitate the electron transfer between redox enzymes and electrodes. Reversible and direct electron transfer of cyt c is successfully achieved at the nanostructured TiO(2) surface with the redox formal potential (E(0)') of 108.0 +/- 1.9 mV versus Ag|AgCl and heterogeneous electron transfer rate constant (k(s)) of 13.8 +/- 2.1 s(-1). Experimental data indicate that cyt c is stably immobilized onto the TiO(2) nanoneedles film and maintains inherent enzymatic activity toward H(2)O(2). On the basis of these results, the cyt c-TiO(2) nanocomposits film is capable of sensing H(2)O(2) at a suitable potential, 0.0 V (vs Ag|AgCl), where not only common anodic interferences like ascorbic acid, uric acid, 3,4 dihydroxyphenylacetic acid but also a cathodic interference, O(2), are effectively avoided. Besides high selectivity, the present biosensor for H(2)O(2) shows broad dynamic range and low detection limit. These remarkable analytical advantages, as well as the characteristic of TiO(2) nanoneedles film such as high conductivity, biocompatibility, and facile ability to miniaturize establishes a novel approach to detection of extracellular H(2)O(2) released from human liver cancer cells. PMID- 19290668 TI - In vitro activity of the EWS oncogene transcriptional activation domain. AB - Aberrant chromosomal fusion of the Ewings sarcoma oncogene (EWS) to several different cellular partners gives rise to the Ewing's family of oncogenic proteins [EWS fusion proteins (EFPs)] and associated tumors (EFTs). EFPs are potent transcriptional activators dependent on the N-terminal region of EWS [the EWS activation domain (EAD)], and this function is thought to be central to EFT oncogenesis and maintenance. Thus, EFPs are promising therapeutic targets, and detailed molecular studies of the EAD will be pivotal for exploring this potential. For many reasons, the molecular mechanism of EAD action is poorly understood and one major obstacle to progress is the lack of an in vitro transcription assay. Using well-characterized EAD-dependent activators and soluble nuclear extracts, we have attempted to recapitulate EAD transcriptional activity in vitro. We report that while the EAD activates transcription strongly in vitro, the effect of EAD mutations is strikingly different from that observed in vivo. Our results therefore suggest that crude soluble extracts do not support bona fide EAD activity in vitro, and we discuss our findings in relation to future assay development and potential mechanisms of EAD action. PMID- 19290669 TI - Direct observation of molecular structural change during intersystem crossing by real-time spectroscopy with a few optical cycle laser. AB - Ultrafast spectroscopy by a sub-5 fs pulse laser was applied to the simultaneous study of electronic relaxation and vibrational dynamics in Ru(II)(TPP)(CO). The electronic lifetimes of (1)Q(x(1,0))(pi,pi*) and (1)Q(x(0,0))(pi,pi*) were determined to be 230 +/- 70 fs and 1150 +/- 260 fs, respectively. The spectrogram shows the time dependent changes in the vibrational spectrum associated with the spin state change from the Franck-Condon state in the excited singlet state to the triplet state via the curve crossing point between the singlet and triplet potential surfaces. The time constant of the intersystem crossing process was determined to be about 1.0 ps from observed electronic relaxation and vibrational dynamics reflecting the transition from the singlet to triplet electronic excited state. PMID- 19290670 TI - Lighting up individual DNA binding proteins with quantum dots. AB - The ability to determine the precise loci and occupancy of DNA-binding proteins is instrumental to our understanding of cellular processes like gene expression and regulation. We propose a single-molecule approach for the direct visualization of proteins bound to their template DNA. Fluorescent quantum dots (QD) are used to label proteins bound to DNA, allowing multicolor, nanometer resolution localization. Protein-DNA complexes are linearly extended and imaged to determine the precise location of the protein binding sites. The method is demonstrated by detecting individual QD-labeled T7-RNA polymerases on the T7 bacteriophage genome. This work demonstrates the potential of this approach to precisely read protein binding position or, alternatively, "write" such information on extended DNA with QDs via sequence-specific molecular recognition. PMID- 19290672 TI - Cellular uptake of platinum nanoparticles in human colon carcinoma cells and their impact on cellular redox systems and DNA integrity. AB - Supercritical fluid reactive deposition was used for the deposition of highly dispersed platinum nanoparticles with controllable metal content and particle size distribution on beta-cyclodextrin. The average particle size and size distribution were steered by the precursor reduction conditions, resulting in particle preparations <20, <100, and >100 nm as characterized by transmission electron microscopy and scanning electron microscopy (SEM). These particle preparations of different size distributions were used to address the question as to whether metallic platinum particles are able to invade cells of the gastrointestinal tract as exemplified for the human colon carcinoma cell line HT29 and thus affect the cellular redox status and DNA integrity. Combined focused ion beam and SEM demonstrated that platinum nanoparticles were taken up into HT29 cells in their particulate form. The chemical composition of the particles within the cells was confirmed by energy-dispersive X-ray spectroscopy. The potential influence of platinum nanoparticles on cellular redoxsystems was determined in the DCF assay, on the translocation of Nrf-2 and by monitoring the intracellular glutathione (GSH) levels. The impact on DNA integrity was investigated by single cell gel electrophoresis (comet assay) including the formation of sites sensitive to formamidopyrimidine-DNA-glycosylase. Platinum nanoparticles were found to decrease the cellular GSH level and to impair DNA integrity with a maximal effect at 1 ng/cm(2). These effects were correlated with the particle size in an inverse manner and were enhanced with increasing incubation time but appeared not to be based on the formation of reactive oxygen species. PMID- 19290671 TI - Regulation of FMN subdomain interactions and function in neuronal nitric oxide synthase. AB - Nitric oxide synthases (NOS) are modular, calmodulin- (CaM-) dependent, flavoheme enzymes that catalyze oxidation of l-arginine to generate nitric oxide (NO) and citrulline. During catalysis, the FMN subdomain cycles between interaction with an NADPH-FAD subdomain to receive electrons and interaction with an oxygenase domain to deliver electrons to the NOS heme. This process can be described by a three-state, two-equilibrium model for the conformation of the FMN subdomain, in which it exists in two distinct bound states (FMN-shielded) and one common unbound state (FMN-deshielded). We studied how each partner subdomain, the FMN redox state, and CaM binding may regulate the conformational equilibria of the FMN module in rat neuronal NOS (nNOS). We utilized four nNOS protein constructs of different subdomain composition, including the isolated FMN subdomain, and determined changes in the conformational state by measuring the degree of FMN shielding by fluorescence, electron paramagnetic resonance, or stopped-flow spectroscopic techniques. Our results suggest the following: (i) The NADPH-FAD subdomain has a far greater capacity to interact with the FMN subdomain than does the oxygenase domain. (ii) CaM binding has no direct effects on the FMN subdomain. (iii) CaM destabilizes interaction of the FMN subdomain with the NADPH FAD subdomain but does not measurably increase its interaction with the oxygenase domain. Our results imply that a different set point and CaM regulation exists for either conformational equilibrium of the FMN subdomain. This helps to explain the unique electron transfer and catalytic behaviors of nNOS, relative to other dual-flavin enzymes. PMID- 19290673 TI - Flexible pores of a metal oxide-based capsule permit entry of comparatively larger organic guests. AB - In zeolites and other rigid solid-state oxides, substrates whose sizes exceed the pore dimensions of the material are rigorously excluded. Now, using a porous 3 nm diameter capsule-like oxomolybdate complex [{Mo(VI)(6)O(21)(H(2)O)(6)}(12){(Mo(V)(2)O(4))(30)(OAc)(21)(H(2)O)(18)}](33-) as a water-soluble analogue of solid-state oxides (e.g., as a soluble analogue of 3 A molecular sieves), we show that carboxylates (RCO(2)(-)) can negotiate passage through flexible Mo(9)O(9) pores in the surface of the capsule and that the rates follow the general trend R = 1 degree >> 2 degrees > 3 degrees >> phenyl (no reaction). Surprisingly, the branched alkanes (R = iso-Pr and tert-Bu) enter the capsule even though they are larger than the crystallographic dimensions of the Mo(9)O(9) pores. Four independent lines of spectroscopic and kinetic evidence demonstrate that these organic guests enter the interior of the capsule through its Mo(9)O(9) apertures and that no irreversible changes in the metal oxide framework are involved. This unexpected phenomenon likely reflects the greater flexibility of molecular versus solid-state structures and represents a sharp departure from traditional models for diffusion through porous solid-state (rigid) oxides. PMID- 19290674 TI - Physical activity in culturally and linguistically diverse migrant groups to Western society: a review of barriers, enablers and experiences. AB - A close examination of epidemiological data reveals burdens of disease particular to culturally and linguistically diverse (CALD) migrants, as these individuals adjust to both culture and modernization gaps. Despite the increased risk of hypertension, diabetes mellitus, overweight/obesity and cardiovascular disease, individuals from CALD groups are less likely to be proactive in accessing healthcare or undertaking preventative measures to ensure optimal health outcomes. The purpose of this paper is to review literature that outlines the barriers, challenges and enablers of physical activity in CALD groups who have recently migrated to Western society, and to identify key strategies to increase physical activity participation for these individuals. Electronic and manual literature searches were used to identify 57 publications that met the inclusion criteria. Findings from the review indicate that migration to Western societies has a detrimental effect on the health status and health behaviours of CALD groups as they assimilate to their new surroundings, explore different cultures and customs, and embrace a new way of life. In particular, there is evidence that physical inactivity is common in migrant CALD groups, and is a key contributing risk factor to chronic disease for these individuals. Challenges and barriers that limit physical activity participation in CALD groups include: cultural and religious beliefs, issues with social relationships, socioeconomic challenges, environmental barriers, and perceptions of health and injury. Strategies that may assist with overcoming these challenges and barriers consist of the need for cultural sensitivity, the provision of education sessions addressing health behaviours, encouraging participation of individuals from the same culture, exploration of employment situational variables, and the implementation of 'Health Action Zones' in CALD communities. This information will inform and support the development of culturally appropriate programmes designed to positively influence the physical activity behaviours of individuals from CALD populations. PMID- 19290675 TI - Physiological differences between cycling and running: lessons from triathletes. AB - The purpose of this review was to provide a synopsis of the literature concerning the physiological differences between cycling and running. By comparing physiological variables such as maximal oxygen consumption (V O(2max)), anaerobic threshold (AT), heart rate, economy or delta efficiency measured in cycling and running in triathletes, runners or cyclists, this review aims to identify the effects of exercise modality on the underlying mechanisms (ventilatory responses, blood flow, muscle oxidative capacity, peripheral innervation and neuromuscular fatigue) of adaptation. The majority of studies indicate that runners achieve a higher V O(2max) on treadmill whereas cyclists can achieve a V O(2max) value in cycle ergometry similar to that in treadmill running. Hence, V O(2max) is specific to the exercise modality. In addition, the muscles adapt specifically to a given exercise task over a period of time, resulting in an improvement in submaximal physiological variables such as the ventilatory threshold, in some cases without a change in V O(2max). However, this effect is probably larger in cycling than in running. At the same time, skill influencing motor unit recruitment patterns is an important influence on the anaerobic threshold in cycling. Furthermore, it is likely that there is more physiological training transfer from running to cycling than vice versa. In triathletes, there is generally no difference in V O(2max) measured in cycle ergometry and treadmill running. The data concerning the anaerobic threshold in cycling and running in triathletes are conflicting. This is likely to be due to a combination of actual training load and prior training history in each discipline. The mechanisms surrounding the differences in the AT together with V O(2max) in cycling and running are not largely understood but are probably due to the relative adaptation of cardiac output influencing V O(2max) and also the recruitment of muscle mass in combination with the oxidative capacity of this mass influencing the AT. Several other physiological differences between cycling and running are addressed: heart rate is different between the two activities both for maximal and submaximal intensities. The delta efficiency is higher in running. Ventilation is more impaired in cycling than in running. It has also been shown that pedalling cadence affects the metabolic responses during cycling but also during a subsequent running bout. However, the optimal cadence is still debated. Central fatigue and decrease in maximal strength are more important after prolonged exercise in running than in cycling. PMID- 19290676 TI - Treatment of common deficits associated with chronic ankle instability. AB - Lateral ankle sprains are amongst the most common injuries incurred by athletes, with the high rate of reoccurrence after initial injury becoming of great concern. Chronic ankle instability (CAI) refers to the development of repetitive ankle sprains and persistent residual symptoms post-injury. Some of the initial symptoms that occur in acute sprains may persist for at least 6 months post injury in the absence of recurrent sprains, despite the athlete having returned to full functional activity. CAI is generally thought to be caused by mechanical instability (MI) or functional instability (FI), or both. Although previously discussed as separate entities, recent research has demonstrated that deficits associated with both MI and FI may co-exist to result in CAI. For clinicians, the main deficits associated with CAI include deficits in proprioception, neuromuscular control, strength and postural control. Based on the literature reviewed, it does seem that subjects with CAI have a deficit in frontal plane ankle joint positional sense. Subjects with CAI do not appear to exhibit any increased latency in the peroneal muscles in response to an external perturbation. Preliminary data suggest that feed-forward neuromuscular control may be more important than feed-back neuromuscular control and interventions are now required to address deficits in feed-forward neuromuscular control. Balance training protocols have consistently been shown to improve postural stability in subjects with CAI. Subjects with CAI do not experience decreased peroneus longus strength, but instead may experience strength deficits in the ankle joint invertor muscles. These findings are of great clinical significance in terms of understanding the mechanisms and deficits associated with CAI. An appreciation of these is vital to allow clinicians to develop effective prevention and treatment programmes in relation to CAI. PMID- 19290677 TI - The anatomy of the pubic region revisited: implications for the pathogenesis and clinical management of chronic groin pain in athletes. AB - Chronic groin pain is a common complaint for athletes participating in sports that involve repetitive sprinting, kicking or twisting movements, such as Australian Rules football, soccer and ice hockey. It is frequently a multifactorial condition that presents a considerable challenge for the treating sports medicine practitioner. To better understand the pathogenesis of chronic groin pain in athletes, a precise anatomical knowledge of the pubic symphysis and surrounding soft tissues is required. Several alternative descriptions of pubic region structures have been proposed. Traditionally, chronic groin pain in athletes has been described in terms of discrete pathology requiring specific intervention. While this clinical reasoning may apply in some cases, a review of anatomical findings indicates the possibility of multiple pathologies coexisting in athletes with chronic groin pain. An appreciation of these alternative descriptions may assist sports medicine practitioners with diagnostic and clinical decision-making processes. The purpose of this literature review is to reappraise the anatomy of the pubic region, considering findings from cadaveric dissection and histology studies, as well as those from diagnostic imaging studies in athletes. PMID- 19290678 TI - Annual age-grouping and athlete development: a meta-analytical review of relative age effects in sport. AB - Annual age-grouping is a common organizational strategy in sport. However, such a strategy appears to promote relative age effects (RAEs). RAEs refer both to the immediate participation and long-term attainment constraints in sport, occurring as a result of chronological age and associated physical (e.g. height) differences as well as selection practices in annual age-grouped cohorts. This article represents the first meta-analytical review of RAEs, aimed to collectively determine (i) the overall prevalence and strength of RAEs across and within sports, and (ii) identify moderator variables. A total of 38 studies, spanning 1984-2007, containing 253 independent samples across 14 sports and 16 countries were re-examined and included in a single analysis using odds ratios and random effects procedures for combining study estimates. Overall results identified consistent prevalence of RAEs, but with small effect sizes. Effect size increased linearly with relative age differences. Follow-up analyses identified age category, skill level and sport context as moderators of RAE magnitude. Sports context involving adolescent (aged 15-18 years) males, at the representative (i.e. regional and national) level in highly popular sports appear most at risk to RAE inequalities. Researchers need to understand the mechanisms by which RAEs magnify and subside, as well as confirm whether RAEs exist in female and more culturally diverse contexts. To reduce and eliminate this social inequality from influencing athletes' experiences, especially within developmental periods, direct policy, organizational and practitioner intervention is required. PMID- 19290679 TI - The importance of cellular phosphorus in controlling the uptake and toxicity of cadmium and zinc in Microcystis aeruginosa, a freshwater cyanobacterium. AB - In the present study, we quantified the 4-h uptake and 48-h toxicity of Cd and Zn in a freshwater cyanobacterium, Microcystis aeruginosa, under varied cellular and ambient P concentrations. After acclimation under different P conditions, the cyanobacteria had different cellular P concentrations. We found that an elevated cellular P concentration significantly promoted the short-term uptake of Cd and Zn by M. aeruginosa. With an increase in the cellular P concentration from 66 to 118 micromol/g, the uptake rates of Cd and Zn increased by 40- and 16-fold, respectively, and a significant exponential relationship between metal uptake rate and cellular P concentration was observed. The pulse amplitude-modulated parameter (maximum photosystem II quantum yield) and cell-specific growth rate were used as toxicity endpoints of cyanobacteria over 48 h of metal exposure. The P-replete cells were more tolerant of [Cd2+] or [Zn2+] than the P-starved cells but became more sensitive to Cd toxicity when incubated in a P-deficient medium. The polyphosphate bodies may have formed in the cyanobacterial cells under surplus P conditions, which can serve as a metal sink to sequester/detoxify the incoming Cd and Zn. The geometric means of median inhibition concentration based on the cellular metal to P ratio (mol:mol) were 0.041 and 0.036 for Cd and Zn, respectively. The cellular metal to P ratio was better than the cellular P concentration at predicting the toxic effects of metals under different P conditions. PMID- 19290680 TI - Toxicity of two insecticides to California, USA, anurans and its relevance to declining amphibian populations. AB - Contaminants have been associated with population declines of several amphibian species in California (USA). Pesticides from the Central Valley of California are transported by winds into the Sierra Nevada Mountains and precipitate into wet meadows where amphibians breed. The present study examined the chronic toxicity of two of the insecticides most commonly used in the Central Valley and found in the mountains, chlorpyrifos and endosulfan, to larval Pacific treefrogs (Pseudacris regilla) and foothill yellow-legged frogs (Rana boylii) and discusses the implications of this toxicity to declining amphibian populations. Larvae were exposed to the pesticides from Gosner stages 25 to 26 through metamorphosis. The estimated median lethal concentration (LC50) for chlorpyrifos was 365 microg/L in P. regilla and 66.5 microg/L for R. boylii. Time to metamorphosis increased with concentration of chlorpyrifos in both species, and cholinesterase activity declined with exposure concentration in metamorphs of both species at Gosner stages 42 to 46. For endosulfan, the estimated LC50 was 15.6 microg/L for P. regilla and 0.55 microg/L for R. boylii. All R. boylii exposed to concentrations of greater than 0.8 microg/L died before they entered metamorphosis. Pseudacris regilla remains relatively abundant and is broadly distributed throughout California. In contrast, R. boylii is among the species experiencing severe population declines. The present study adds to the increasing evidence that pesticides are very harmful to amphibians living in areas that are miles from sources of pesticide application. PMID- 19290681 TI - Application of a stable isotope technique to determine the simultaneous uptake of cadmium, copper, nickel, lead, and zinc by the water flea Daphnia magna from water and the green algae Pseudokirchneriella subcapitata. AB - Accumulation and toxicological effects of water and dietary metals in aquatic organisms can potentially be very different. Therefore, it is important to know the relative contribution of these different sources to metal exposure, availability, and accumulation. In the present study, a stable isotope technique was applied to investigate the uptake of Cd, Cu, Ni, Pb, and Zn by the green alga Pseudokirchneriella subcapitata and the water flea Daphnia magna during simultaneous exposure to the five metals at environmentally realistic concentrations from separate water and dietary routes. Green algae take up Cu faster compared to Cd, Ni, Pb, and Zn, and the distribution of metals between the external and internal compartments is dependent on metal and population growth stage. The metal accumulation reached a steady state within 24 to 48 h for all metals. Metal uptake rate constants from water were highest for Cu and lowest for Ni. Metal assimilation efficiencies from the food source varied with metal, ranging from approximately 80% in the case of Cd to near 0% in the case of Ni. Because the data for the different metals were obtained on the same multimetal exposed organisms, the results are directly comparable among the metals. For all five metals studied, water appeared to be the most important route of uptake by D. magna. PMID- 19290682 TI - Effects of dietary N-acetylcysteine on the oxidative stress induced in tilapia (Oreochromis niloticus) exposed to a microcystin-producing cyanobacterial water bloom. AB - Fish can be exposed to toxic cyanobacterial cells in natural waters and fish farms and suffer from oxidative damage. The present study investigates the effects of N-acetylcysteine (NAC), a glutathione (GSH) precursor, on the oxidative stress induced by Microcystis cyanobacterial cells containing microcystins (MCs) in tilapia fish (Oreochromis niloticus). Variation in lipid peroxidation (LPO) levels, carbonyl group content, reduced glutathione to oxidized glutathione ratio (GSH:GSSG), and catalase (Enzyme Commission [EC] 1.11.1.6), superoxide dismutase (SOD; EC 1.15.1.1), glutathione reductase (GR; EC 1.8.1.7), glutathione peroxidase (GPx; EC 1.11.1.9), and glutathione S transferase (EC 2.5.1.18) activities in liver and kidney of tilapia exposed to a single oral dose of 120 microg MC-LR (with leucine [L] and arginine [R])/fish and killed in 24 h were investigated in the absence and presence of 20.0, 44.0, and 96.8 mg NAC/fish/d. Results showed a protective role of NAC, depending on the dose and the biomarker considered. The increase in LPO (1.9- and 1.4-fold in liver and kidney, respectively) and the decreased protein content and GSH: GSSG in the liver induced by MCs were recovered mainly by the lower doses of NAC employed. Antioxidant enzyme activities increased (range, 1.4- to 1.7-fold) by MCs also were ameliorated by NAC, although the highest level used induced significant alteration of some enzymatic activities, such as SOD, GPx, and GR. Thus, NAC can be considered to be a useful chemoprotectant that reduces hepatic and renal oxidative stress in the prophylaxis and treatment of MC-related intoxications in fish when careful attention is given to its application dose because of its own pro-oxidant activity, as shown in the present study at 96.8 mg NAC/fish/d. PMID- 19290683 TI - Reversibility of estrogenic sex changes in zebrafish (Danio rerio). AB - Development of male zebrafish (Danio rerio) courtship behavior was studied following estrogenic disruption of sexual differentiation. Sixty zebrafish were exposed at 28 degrees C to 5 ng/L (nominal concentration) of 17alpha ethinylestradiol (EE2) from the egg stage until adulthood at four months of age, resulting in a female-biased sex ratio. Twenty-five EE2-exposed phenotypic female zebrafish were subsequently held in clean water for eight months. During this period, eight phenotypic males developed. These phenotypic males demonstrated significant behavioral aberrations and a low fertilization rate compared to control males. PMID- 19290684 TI - Inaugural editorial. AB - Gives a brief history of the Psychology of Addictive Behaviors, noting its current premier status among addictions journals. The editor discusses the types of manuscripts that the journal publishes, covering a wide range of topics and substantive areas. The plan is to keep the journal as an outlet for publication of manuscripts concerning all of the addictive behaviors. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290685 TI - Longitudinal relations between marital aggression and alcohol problems. AB - Relations between couples' (N = 158) marital aggression and alcohol problems were examined across a two-year period. Alcohol problems and aggression were assessed via self-report and partner-reports. Results support bidirectional relations between marital aggression and problem drinking. T1 wife problem drinking was associated with decreased T2 verbal aggression; T1 husband problem drinking was associated with increased T2 physical aggression. T1 physical aggression predicted increased T2 wife problem drinking; it predicted increased T2 husband problem drinking only when wife problem drinking was low. T1 verbal marital aggression predicted increased T2 husband problem drinking only when husbands engaged in greater problem drinking at T1. Results suggest that problem drinking may prevent couples from adequately handling marital disagreements, and that marital problems may lead to drinking as a form of coping with stress; couples in which the husband engages in greater problem drinking than the wife may be at increased risk. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290686 TI - Influences of sexual sensation seeking, alcohol consumption, and sexual arousal on women's behavioral intentions related to having unprotected sex. AB - This experimental study examined effects of alcohol consumption and sexual sensation seeking on unprotected sex intentions, taking into account sexual arousal, indirectly discouraging sex, and condom insistence. Women (N = 173; mean age = 25.02) were randomly assigned to a control, placebo, low-dose beverage (target blood alcohol level = .04), or high- dose beverage (target blood alcohol level = .08) condition. Participants projected themselves into a hypothetical sexual interaction with a man in which no condom was available. Structural equation modeling demonstrated that both sexual sensation seeking and alcohol dose directly increased sexual arousal early in the interaction, but later sexual arousal indirectly increased unprotected sex intentions by decreasing endorsement of indirect discouragement and, in turn, condom insistence. These findings help to clarify the role of alcohol consumption and sensation seeking in women's sexual decision making and point to the importance of examining it as a multistage process. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290687 TI - Movie exposure to alcohol cues and adolescent alcohol problems: a longitudinal analysis in a national sample. AB - The authors tested a theoretical model of how exposure to alcohol cues in movies predicts level of alcohol use (ever use plus ever and recent binge drinking) and alcohol-related problems. A national sample of younger adolescents was interviewed by telephone with 4 repeated assessments spaced at 8-month intervals. A structural equation modeling analysis performed for ever-drinkers at Time 3 (N = 961) indicated that, controlling for a number of covariates, movie alcohol exposure at Time 1 was related to increases in peer alcohol use and adolescent alcohol use at Time 2. Movie exposure had indirect effects to alcohol use and problems at Times 3 and 4 through these pathways, with direct effects to problems from Time 1 rebelliousness and Time 2 movie exposure also found. Prospective risk promoting effects were also found for alcohol expectancies, peer alcohol use, and availability of alcohol in the home; protective effects were found for mother's responsiveness and for adolescent's school performance and self-control. Theoretical and practical implications are discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290688 TI - A longitudinal study of social competence among children of alcoholic and nonalcoholic parents: role of parental psychopathology, parental warmth, and self regulation. AB - This study tested a conceptual model predicting children's social competence in a sample of children with alcoholic and non-alcoholic parents. The model examined the role of parents' alcohol diagnoses, depression, and antisocial behavior at 12 18 months of child age in predicting parental warmth/sensitivity at 2 years of child age. Parental warmth/sensitivity at 2 years was hypothesized to predict children's self-regulation and externalizing behavior problems at 3 years. Parenting, self-regulation, and behavior problems were expected to predict social competence in kindergarten. Structural equations modeling was supportive of this model. Fathers' alcohol diagnosis was associated with lower warmth/sensitivity. Lower maternal warmth/sensitivity at 2 years was predictive of lower child self regulation at 3 years. Parenting, self-regulation, and externalizing behavior problems were predictive of social competence in kindergarten, although associations varied by reporter (parents or teacher). There was a direct association between fathers' alcohol diagnosis and father reports of social competence, and between fathers' depression and teacher reports of social competence. The study elucidates developmental processes in predicting social competence and the role of fathers' alcoholism and associated risk factors in this process. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290689 TI - Betting on change: modeling transitional probabilities to guide therapy development for opioid dependence. AB - This study investigated the process of change by modeling transitions among four clinical states encountered in 64 detoxified opiate-dependent individuals treated with daily oral naltrexone: no opiate use, blocked opiate use (i.e., opiate use while adhering to oral naltrexone), unblocked opiate use (i.e., opiate use after having discontinued oral naltrexone), and treatment dropout. The effects of baseline characteristics and two psychosocial interventions of differing intensity, behavioral naltrexone therapy (BNT) and compliance enhancement (CE), on these transitions were studied. Participants using greater quantities of opiates were more likely than other participants to be retained in BNT relative to CE. Markov modeling indicated a transition from abstinence to treatment dropout was approximately 3.56 times greater among participants in CE relative to participants in BNT, indicating the more comprehensive psychosocial intervention kept participants engaged in treatment longer. Transitions to stopping treatment were more likely to occur after unblocked opiate use in both treatments. Continued opiate use while being blocked accounted for a relatively low proportion of transitions to abstinence and may have more deleterious effects later in a treatment episode. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290690 TI - Self-efficacy and smoking cessation: a meta-analysis. AB - According to relapse models, self-efficacy (SE), or confidence in one's ability to abstain, should predict the outcome of an attempt to quit smoking. We reviewed 54 studies that prospectively examined this relationship. The relationship between SE and future smoking depended upon the population studied and the timing of the SE assessment. The relationship between SE and future smoking was modest when SE was assessed prior to a quit attempt; SE scores were .21 standard deviation units (SD) higher for those not smoking at follow-up than for those who were smoking. The relationship was stronger (.47 SD) when SE was assessed post quit. However, this effect was diminished when only abstainers at the time of the SE assessment were included in analysis (.28 SD). Controlling for smoking status at the time of SE assessment substantially reduced the relationship between SE and future smoking. Although SE has a reliable association with future abstinence, it is less robust than expected. Many studies may overestimate the relationship by failing to appropriately control for smoking behavior at the time of the SE assessment. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290691 TI - Hostility and smoking cessation treatment outcome in heavy social drinkers. AB - Hostility is a multifaceted construct encompassing affective, behavioral, and cognitive aspects. There is preliminary evidence linking hostility to poorer outcomes in smoking cessation treatment; however, it is unclear which components of hostility are most important in cessation. In this study, the authors examined multiple aspects of trait hostility in 92 heavy social drinkers who were seeking smoking cessation treatment. Consistent with their hypothesis, the authors found that the cognitive component of hostility was most relevant to smoking cessation outcome. Specifically, those who expressed bitterness about their lives and tended to believe that they had poor luck and had gotten a raw deal out of life had poor smoking cessation outcomes. Cognitive measures of hostility also predicted greater nicotine withdrawal symptoms 1 week after quitting smoking. Other components of hostility including anger and both physical and verbal aggression did not significantly predict smoking outcome or nicotine withdrawal. Further examination of how a hostile worldview contributes to smoking cessation failure is warranted, as this facet of hostility may prove a valuable target for smoking cessation interventions. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290692 TI - Attention bias in nicotine withdrawal and under stress. AB - Drug motivation models postulate that attention biasing toward smoking-related cues is a cognitive mechanism supporting continued or renewed drug use, and they predict that drug use history, deprivation, and distress should modulate the extent of this bias. The present study used the modified Stroop paradigm to extend past research regarding attention biasing toward smoking and unpleasant, pleasant, and neutral words among adult nonsmokers and daily smokers. Both nonsmokers and smokers showed differential attention toward unpleasant and pleasant cues, particularly pleasant cues, but did not show a unique bias toward smoking-related stimuli. Results suggested that, among smokers, nicotine deprivation and exogenous stress (threat of electric shock) have a nonadditive effect on attention toward pleasant cues but no effect on attention to smoking cues specifically. Similarly, instructing smokers that they would have an opportunity to smoke did not significantly increase the bias of nicotine-deprived smokers' attention toward smoking-related cues, relative to arousing unpleasant and pleasant cues. Overall, results suggest that smokers' attention may be biased toward both smoking-related and other salient cues when deprived of nicotine and anticipating an opportunity to smoke. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290693 TI - Cross-lagged links among gambling, substance use, and delinquency from midadolescence to young adulthood: additive and moderating effects of common risk factors. AB - The authors examined cross-lagged links among gambling, substance use, theft, and violence from midadolescence to young adulthood and whether behavioral disinhibition, deviant peers, and parental supervision as common risk factors explain or moderate those links. In 2 community samples, male Caucasians were assessed for gambling participation and problems with the South Oaks Gambling Screen-Revised for Adolescents (K. C. Winters, R. Stinchfield, & J. Fulkerson, 1993) at age 16 years and the South Oaks Gambling Screen (H. R. Lesieur & S. B. Blume, 1987) at age 23. Other problem behaviors were also assessed both times. Risk factors were measured at age 16. Adolescent substance use was related to subsequent theft and violence but not gambling. Gambling problems were linked to subsequent gambling participation. For adolescents with deviant peers, gambling problems were linked to subsequent theft; this was not the case for adolescents without deviant peers. Only for individuals high on disinhibition did stability of gambling problems resemble moderate stabilities of other problem behaviors. Each risk factor was related to each problem behavior (exception: parenting unrelated to gambling). These risk factors partly explained the cross-lagged links among behaviors and thus may be useful targets of prevention. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290694 TI - The association of form of gambling with problem gambling among American youth. AB - A random telephone survey was conducted with 2,274 United States residents aged 14 to 21. Analyses were performed to assess the relationship between the specific gambling games played and the extent of problem gambling symptoms. The forms of gambling that were most associated with gambling problems were card games, casino gambling, "other" gambling on routine activities, and betting on games of skill such as basketball, pool, or golf. The form of gambling that made the largest contribution to gambling problems per 14 days of play was casino gambling. The hypothesis that rapid forms of gambling, such as slot machines, would be the most problematic forms of gambling was not upheld. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290695 TI - Correlates of gambling among youth in an inner-city emergency department. AB - Correlates of past year gambling were examined in a diverse sample of 1128 youth ages 14 to 18 (54.1% female, 58.0% African American) presenting to an inner-city emergency department (ED). Overall, 22.5% of the sample reported past-year gambling. Male youth were more likely to gamble than female youth, and African American youth reported higher rates of past-year gambling than non-African American youth. Significant bivariate correlates of gambling included lower academic achievement, being out of school, working more than 20 hours per week, alcohol, cigarette, and marijuana use, alcohol problems, severe dating violence, moderate and severe general violence, and carrying a weapon. When examined simultaneously, being male, African American, out of school, working for pay, alcohol and marijuana use, severe general violence, and carrying a weapon all emerged as significant correlates of past-year gambling, largest amount of money gambled, and gambling frequency. In addition, involvement in severe dating violence was associated with frequency and largest amount gambled. The results suggest that gambling is common among youth in the inner city and is associated with several risk behaviors. The inner-city ED may provide a context for screening and intervention to address multiple risk behaviors. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290696 TI - Strength of commitment language in motivational interviewing and gambling outcomes. AB - The purpose of the present study was to examine the mechanisms by which the motivational interview (MI) is an effective treatment for pathological gambling. Forty MIs with pathological gamblers were transcribed and coded for language content and strength of expression. As hypothesized, participants who expressed stronger commitment to change their gambling behavior during the MI exhibited better gambling outcomes over 12 months than did those who expressed weaker commitment or no commitment to change their gambling behavior during the MI. Contrary to expectation, commitment strength in the latter part of the MI was not a stronger predictor of gambling outcome. Expression of desire, ability, need, reasons, and readiness for change were not predictive of outcome. Ability and readiness were associated with commitment. This study has important implications for clinical monitoring of client treatment success and for improving the MI. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290697 TI - Eating behavior in response to food-cue exposure: examining the cue-reactivity and counteractive-control models. AB - Many studies have demonstrated that those high in weight-related concerns eat more after food-cue exposure, which is consistent with predictions of the cue reactivity model. However, the counteractive-control model predicts that exposure to fattening foods activates dieting-related goals and behavior in weight concerned individuals. Although these models seem incongruous, the authors hypothesized that the salience of the cue could represent a critical factor in determining which model is activated. The authors predicted that attending to salient food cues would result in increased intake (cue reactivity) in individuals with high weight-related concerns, whereas incidental food-cue exposure would result in decreased intake (counteractive control), relative to control exposure. The authors employed a 3 (attended vs. incidental vs. control cue) x 2 (low vs. high weight-related concerns) design. As expected, participants with high weight-related concerns who attended to a food cue ate more than did both those with high weight-related concerns in the control condition and those with low weight-related concerns in the attended-cue condition; however, intake of individuals with high weight-related concerns who were exposed to the incidental cue did not differ from that of those in the control condition. The manner of food-cue presentation may be a critical factor in determining eating behavior. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290698 TI - Personality factors and styles among college students who binge eat and drink. AB - Elevated rates of comorbidity between binge eating and alcohol use problems have been widely documented. Prior studies have examined specific personality traits associated with the co-occurrence of these problems. The current study explores comprehensive personality factors that are associated with the co-occurrence of binge eating and binge drinking among a diverse sample of 208 college undergraduates. Using the Five Factor Model of personality, the authors assessed both comprehensive personality factors and style of impulse control, a personality style defined by different combinations of neuroticism and conscientiousness. On the basis of responses to a screening instrument, college students were assigned to one of four groups: binge eat, binge drink, binge eat and drink, and non-binge. The binge eat and drink group reported a higher level of neuroticism than did students in the binge drink and non-binge groups. Additionally, the binge eat and drink group was more likely to report an undercontrolled style of impulse control than were other groups. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290699 TI - Influence of motivational interviewing on explicit and implicit alcohol-related cognition and alcohol use in at-risk adolescents. AB - Both implicit and explicit cognitions play an important role in the development of addictive behavior. This study investigated the influence of a single-session motivational interview (MI) on implicit and explicit alcohol-related cognition and whether this intervention was successful in consequently decreasing alcohol use in at-risk adolescents. Implicit and explicit alcohol-related cognitions were assessed at pretest and one month posttest in 125 Dutch at-risk adolescents ranging in age from 15 to 23 (51 males) with adapted versions of the Implicit Association Test (IAT) and an expectancy questionnaire. Motivation to change, alcohol use and alcohol-related problems were measured with self-report questionnaires, at pretest, at posttest after one month, and at the six-month follow-up. Although the quality of the intervention was rated positively, the results did not yield support for any differential effects of the intervention on drinking behavior or readiness to change at posttest and six-month follow-up. There were indications of changes in implicit and explicit alcohol-related cognitions between pretest and posttest. Our findings raise questions regarding the use of MI in this particular at-risk adolescent population and the mechanisms through which MI is effective. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290700 TI - Measured alcohol content in college party mixed drinks. AB - The main objective of this study was to measure the alcohol content in college party drinks. Samples of mixed drinks were collected from on-campus parties (N = 23) over a 12-week period at a university in the Northeast. Samples were analyzed by using a method that measures oxygen utilization during ethanol oxidation. Standard drink equivalents were calculated and blood alcohol concentrations (BACs) for men and women were estimated. The percent alcohol in sampled drinks ranged from 3.7% to 22.8%. Characteristics of the sampled parties were not related to drink concentration. A party drink at the median concentration and drink size contained 0.97 standard drinks. Estimated BACs varied widely depending on drink alcohol concentration, but in most cases a heavy drinking episode for both men and women resulted in an estimated BAC at or above .08. Mixed drinks at the sampled parties on average approximated one standard drink, but the variability in mixed drink strength compromises a drinker's ability to keep track of the number of drinks consumed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290701 TI - What men want: the role of reflective opposite-sex normative preferences in alcohol use among college women. AB - Misperceptions of peer drinking norms have been found to be strongly associated with individual drinking behavior, especially for proximal reference groups such as same-sex friends. Less studied are the effects of perceived preferences from the opposite sex on alcohol use; that is, the behaviors an individual believes the opposite sex prefers from them. Research suggests that these perceived "reflective" normative preferences may be particularly salient among college women, who may drink in pursuit of intimate relationships and positive attention from male peers. Heterosexual undergraduate students from two universities participated in this project. Females answered questions regarding the amount of alcohol they believe a typical male would like his female friends, dates, or romantic partners to drink. Males answered the same questions, stating their actual preferences. Results showed that females overestimate the amount of alcohol males want their female friends, dating partners, and sexual partners to drink, and that this misperception was associated with their drinking behavior, even after controlling for perceived same-sex norms. These results suggest that reflective normative feedback may offer a powerful new tool for female-targeted interventions. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290702 TI - Brief alcohol intervention with college student drinkers: face-to-face versus computerized feedback. AB - Research has demonstrated that brief interventions featuring personalized feedback can be used to decrease alcohol use among heavy-drinking college students. The current study investigated the efficacy of face-to-face and computer delivered interventions relative to an assessment-only control condition. The content of the personalized feedback was identical across the face to-face and computerized conditions. There were 84 at-risk students assessed before, and 4 weeks after, the delivery of the interventions. The results suggest that both face-to-face and computerized interventions were equally successful in reducing the quantity and frequency of alcohol consumption, and that both interventions were more effective than the control condition. Participants also rated both interventions as acceptable, although the face-to-face intervention was given a more favorable rating. These initial results suggest that computerized interventions can be used to efficiently reduce alcohol use among college students. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290703 TI - Comparable efficacy of contingency management for cocaine dependence among African American, Hispanic, and White methadone maintenance clients. AB - Cocaine use is a significant problem among methadone maintenance clients. Contingency management (CM) is a reinforcement-based approach with demonstrated efficacy for reducing cocaine use. This study examines whether the efficacy of CM treatment for cocaine-dependent individuals receiving methadone maintenance for opioid dependence differs by ethnicity. Participants were 191 African American, Hispanic, and White cocaine-dependent methadone maintenance clients, randomly assigned to standard methadone treatment or standard methadone treatment plus CM for 12 weeks. Hispanic participants were younger, less educated, and reported fewer years of cocaine use than did African American and White participants and reported fewer years of heroin use than did African American participants. African American participants were less likely to report a history of psychiatric symptoms or treatment in comparison with Hispanic and White participants. While CM was associated with longer duration of continuous cocaine abstinence and a greater proportion of submitted urine samples negative for cocaine, ethnicity was not related to treatment outcomes, and there was no significant interaction between treatment and ethnicity. CM appears to be an efficacious treatment for cocaine dependence among methadone maintenance clients, regardless of ethnicity. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290704 TI - Disclosure of sexual orientation and subsequent substance use and abuse among lesbian, gay, and bisexual youths: critical role of disclosure reactions. AB - Research on whether disclosure of sexual orientation promotes lower substance use among lesbian, gay, and bisexual (LGB) individuals has been inconsistent. One reason for this may be that disclosure results in accepting and rejecting reactions. The current report longitudinally examines whether the types of reactions to disclosure are associated with substance use and abuse among 156 LGB youths (ages 14-21). Neither the number of disclosures nor the numbers of accepting or neutral disclosure reactions were associated with substance use or abuse. However, the number of rejecting reactions to disclosure was associated with current and subsequent alcohol, cigarette, and marijuana use, even after controlling for demographic factors, social desirability, and emotional distress. Further, accepting reactions were found to moderate or protect youths from the negative role of rejecting reactions on alcohol use, but not other substances. This research indicates that, rather than disclosure per se, it is the number of accepting and rejecting reactions in response to disclosure that are critical to understanding substance use among LGB youths. Further, the results suggest that to be maximally effective, substance use prevention and treatment efforts should address rejecting reactions. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290705 TI - Influence of stressors on breast cancer incidence in the Women's Health Initiative. AB - OBJECTIVE: To examine associations among life events stress, social support, and breast cancer incidence in a cohort of postmenopausal women. DESIGN AND MAIN OUTCOME MEASURE: Women's Health Initiative observational study participants, breast cancer free at entry, who provided assessment of stressful life events, social support, and breast cancer risk factors, were prospectively followed for breast cancer incidence (n = 84,334). RESULTS: During an average of 7.6 years of follow-up, 2,481 invasive breast cancers were diagnosed. In age-adjusted proportional hazards models, 1 stressful life event was associated with increased risk, but risk decreased with each additional stressful life event. After adjustment for confounders the decreasing risk was not significant. Stressful life events and social support appeared to interact in relation to breast cancer risk such that women who had greater number of stressful life events and low social support had a decreased risk of breast cancer. CONCLUSIONS: This study found no independent association between stressful life events and breast cancer risk. The results are compatible with a more complex model of psychosocial factors interacting in relation to breast cancer risk. PMID- 19290706 TI - Psychosocial adjustment among cancer survivors: findings from a national survey of health and well-being. AB - OBJECTIVE: The current study examined whether cancer survivors showed impairment, resilience, or growth responses relative to a sociodemographically matched sample in four domains: mental health and mood, psychological well-being, social well being, and spirituality. The impact of aging on psychosocial adjustment was also investigated. DESIGN: Participants were 398 cancer survivors who were participants in the MIDUS survey (Midlife in the United States) and 796 matched respondents with no cancer history. Psychosocial assessments were completed in 1995-1996 and 2004-2006. MAIN OUTCOME MEASURES: Outcomes including self-report measures of mental health and mood, psychological well-being, social well-being, and spirituality. RESULTS: Findings indicated that cancer survivors demonstrated impairment relative to the comparison group in mental health, mood, and some aspects of psychological well-being. Longitudinal analyses spanning pre- and postdiagnosis clarified that while mental health declined after a cancer diagnosis, poorer functioning in other domains existed prior to diagnosis. However, survivors exhibited resilient social well-being, spirituality, and personal growth. Moreover, age appeared to confer resiliency; older survivors were more likely than younger adults to show psychosocial functioning equivalent to their peers. CONCLUSION: While younger survivors may be at risk for disturbances in mental health and mood, cancer survivors show resilience in other important domains of psychosocial adjustment. PMID- 19290707 TI - Major depressive disorder is associated with attenuated cardiovascular reactivity and impaired recovery among those free of cardiovascular disease. AB - OBJECTIVE: To examine cardiovascular reactivity and recovery to laboratory stress among a naturalistic sample of individuals diagnosed with major depressive disorder (MDD) and healthy control participants. Prospective evidence suggests that MDD confers risk for cardiovascular disease equal to or greater than the risk associated with depressed mood. Enhanced cardiovascular reactivity has been proposed as a mechanism explaining increased risk, but data are inconsistent as to whether depressed individuals exhibit enhanced or attenuated reactivity. Further, few studies have examined appraisal and recovery differences. DESIGN: Participants diagnosed with MDD (N = 25) and healthy control participants (N = 25) engaged in a cardiovascular reactivity protocol including 2 tasks, each followed by a brief recovery period. MAIN OUTCOME MEASURES: Blood pressure, heart rate, pre-ejection period, cardiac output and total peripheral resistance were assessed. Appraisals of tasks were assessed prior to each task. RESULTS: Depressed participants exhibited significantly less systolic blood pressure, heart rate, and cardiac output reactivity during speech, less heart rate reactivity during mirror tracing, and less heart rate recovery after speech and mirror tracing than controls. Depressed participants appraised the tasks as more demanding, threatening, and stressful and reported being less able to cope than controls. Appraisals were related to heart rate reactivity, but appraisals did not mediate the relationship between depression group and reactivity. CONCLUSION: Impaired recovery rather than exaggerated cardiovascular reactivity may partially explain the increased prospective cardiovascular disease risk in depressed individuals. PMID- 19290708 TI - Self-efficacy as a marker of cardiac function and predictor of heart failure hospitalization and mortality in patients with stable coronary heart disease: findings from the Heart and Soul Study. AB - OBJECTIVE: The authors sought to evaluate the association of self-efficacy with objective measures of cardiac function, subsequent hospitalization for heart failure (HF), and all-cause mortality. DESIGN: Observational cohort of ambulatory patients with stable CHD. The authors measured self-efficacy using a published, validated, 5-item summative scale, the Sullivan Self-Efficacy to Maintain Function Scale. The authors also performed a cardiac assessment, including an exercise treadmill test with stress echocardiography. MAIN OUTCOME MEASURES: Hospitalizations for HF, as determined by blinded review of medical records, and all-cause mortality, with adjustment for demographics, medical history, medication use, depressive symptoms, and social support. RESULTS: Of the 1,024 predominately male, older CHD patients, 1013 (99%) were available for follow-up, 124 (12%) were hospitalized for HF, and 235 (23%) died during 4.3 years of follow up. Mean cardiac self-efficacy score was 9.7 (SD 4.5, range 0-20), corresponding to responses between "not at all confident" and "somewhat confident" for ability to maintain function. Lower self-efficacy predicted subsequent HF hospitalization (OR per SD decrease = 1.4, p = .0006), and all-cause mortality (OR per SD decrease = 1.4, p < .0001). After adjustment, the association of cardiac self efficacy with both HF hospitalization and mortality was explained by worse baseline cardiac function. CONCLUSION: Among patients with CHD, self-efficacy was a reasonable proxy for predicting HF hospitalizations. The increased risk of HF associated with lower baseline self-efficacy was explained by worse cardiac function. These findings indicate that measuring cardiac self-efficacy provides a rapid and potentially useful assessment of cardiac function among outpatients with CHD. PMID- 19290709 TI - Evaluating three theory-based interventions to increase physicians' recommendations of smoking cessation services. AB - OBJECTIVE: To evaluate three theory-based interventions aimed at increasing the rate at which primary care physicians recommend smoking cessation services to smokers. DESIGN: Primary care physicians (n = 251) were randomized to one of four conditions: (a) information based on social cognitive theory (SCT) targeting outcome expectations, (b) information based on SCT + elaboration likelihood model, (c) feedback based on self-perception theory, or (d) control. MAIN OUTCOME MEASURES: Intentions to recommend and self-reported recommendations of smoking cessation services 1 week postintervention. RESULTS: Data were analyzed using covariance and mean structure analysis. Compared with the control group, only the SCT group reported more frequently recommending services (mean difference = 1.1 recommendations per week, Cohen's d = 0.46) and higher intentions. Mediation analysis was consistent with increased intentions resulting from changes in outcome expectations. There was no evidence that changes in intentions explained self-reported behavior change. CONCLUSION: The study provides preliminary "proof of principle" for further studies incorporating more robust outcome measures. PMID- 19290710 TI - Validity of stage assessment in the adoption and maintenance of physical activity and fruit and vegetable consumption. AB - OBJECTIVE: Stage assessments are examined to develop and test refined measurements that can be used for classifying individuals. DESIGN: Stages were assessed in 1,850 persons in terms of their physical activity and dietary behaviors. MAIN OUTCOME MEASURES: Stages for both behaviors were compared to behavior and other test variables. Misclassification, sensitivity, specificity, receiver-operation-curves, and discontinuity patterns were computed. Discontinuity patterns were tested with trends across stages and planned contrasts between adjacent stages. RESULTS: In comparison to previous studies, sensitivity (70% to 80%) and specificity (80% to 87%) were high. When using lower level criteria (such as less intensive activity), sensitivity was lower, whereas specificity was higher. When behavioral maintenance was assessed, results suggested that the temporal cut-off point between action and maintenance was equally optimal at different cut-off points. Applying contrast analyses, nonlinear trends across the stages and a match of 87% of predictions of stage differences resulted. CONCLUSION: Stage assumptions are supported in general, and refined stage assessment in particular. Levels of psychological variables (e.g., easiness, habit) may discriminate stages as well as or even better than temporal stage definitions. PMID- 19290711 TI - A real-time assessment of work stress in physicians and nurses. AB - OBJECTIVE: This study adapted ecological momentary assessment methods to: (a) examine differences in work stress between nurses and physicians, and (b) to study relationships between work stress, work activity patterns, and sleep. DESIGN: A total of 185 physicians and 119 nurses (206 women, 98 men) working in four teaching hospitals participated in an observational study of work stress. MAIN OUTCOME MEASURES: Participants carried handheld computers that randomly prompted them for work activity, patient load, and work stress information. RESULTS: Participants completed more than 9,500 random interval surveys during the study (an average of 30.8 surveys per person-week). Approximately 85% of all surveys were completed in full (73.3%) or partially (11.6%). Emotional stress scores among physicians were nearly 50% higher (26.9[19.0]) than those of nurses (18.1[14.9], r[302] = .37, p < .001). Direct and indirect care activities were associated with higher stress reports by both clinician groups (rs[159] = .14 .26, ps < .01). Sleep quality and quantity were predictors of work stress scores (ps < .05). Finally, higher work stress and lower sleep quality were also associated with poorer memory performance (r[302] = -.12, .17, ps < 05). CONCLUSIONS: The findings identify patterns of work stress in relationship to work activities, sleep habits, and provider differences that may be used to assist ongoing hospital work reform efforts. PMID- 19290712 TI - Explanations for side effect aversion in preventive medical treatment decisions. AB - OBJECTIVE: Many laypeople demonstrate excessive sensitivity to negative side effects of medical treatments, which may lead them to refuse beneficial therapies. This Internet-based experiment investigated three possible explanations for such "side effect aversion." One was derived from mental accounting, one examined the mere presence of a side effect, and one focused on computational difficulties. DESIGN: Participants (N = 5,379) were presented with a hypothetical cancer preventive treatment situation that was or was not accompanied by one or two small side effects. The side effects were either beneficial or harmful. In all conditions, the net absolute risk reduction associated with the treatment was 15%. MAIN OUTCOME MEASURES: Participants indicated their willingness to accept treatment and their perceptions of the treatment's effects on their overall cancer risk. RESULTS: Data were consistent only with the "mere presence" explanation of side effect aversion, the idea that side effects act as a strong negative cue that directly affects treatment appraisal. The number of negative side effects did not influence treatment willingness. CONCLUSION: Side effect aversion is a challenge to informed decision making. Specific mechanisms that produce side effect aversion should be identified. PMID- 19290713 TI - Using icon arrays to communicate medical risks: overcoming low numeracy. AB - OBJECTIVE: Icon arrays have been suggested as a potentially promising format for communicating risks to patients-especially those with low numeracy skills-but experimental studies are lacking. This study investigates whether icon arrays increase accuracy of understanding medical risks, and whether they affect perceived seriousness of risks and helpfulness of treatments. DESIGN: Two experiments were conducted on samples of older adults (n = 59, 62 to 77 years of age) and university students (n = 112, 26 to 35 years of age). MAIN OUTCOME MEASURES: Accuracy of understanding risk reduction; perceived seriousness of risks; perceived helpfulness of treatments. RESULTS: Icon arrays increased accuracy of both low- and high-numeracy people, even when transparent numerical representations were used. Risks presented via icon arrays were perceived as less serious than those presented numerically. With larger icon arrays (1,000 instead of 100 icons) risks were perceived more serious, and risk reduction larger. CONCLUSIONS: Icon arrays are a promising way of communicating medical risks to a wide range of patient groups, including older adults with lower numeracy skills. PMID- 19290714 TI - Reconceptualizing decisional balance in an adolescent sun protection intervention: mediating effects and theoretical interpretations. AB - OBJECTIVE: The Transtheoretical model (TTM) integrates principles of operant learning, such as stimulus control and reinforcement, and psychological factors, such as decisional balance. Understanding interrelationships between decisions, behavior, and consequences from multiple-theoretical perspectives can advance theory and inform development of more effective interventions. METHOD: This analysis examined the mediating effects of a special case of the decisional balance construct in which the pros of competing behaviors (i.e., sun protection vs. exposure) were measured rather than the pros and cons of the same behavior. Participants included 819 adolescents (10 to 16 years old, 53.5% girls, 58.4% White) randomized to a 24-month expert system intervention (SunSmart) or a physical activity and nutrition comparison group. MAIN OUTCOME MEASURES: Self report measures included sun protection behaviors, pros for protection, and pros for exposure. Mediation analysis using latent growth curve models found both the treatment-to-mediator and mediator-to-behavior paths significant for decisional balance, producing an indirect effect of .323 (p < .01) and good model fit (CFI = .973, RMSEA = .055). RESULTS: Multiple strategies for conceptualizing and measuring decisional balance appear to be valid. Results are interpreted from the TTM and operant perspectives. PMID- 19290715 TI - Symptom perception in children with asthma: cognitive and psychological factors. AB - OBJECTIVE: This study tested the differential effects of several cognitive and psychological variables on children's perception of asthma symptoms by use of an Asthma Risk Grid. Children's subjective and objective assessments of PEFR (peak expiratory flow rate) were characterized as representing perceptual accuracy, symptom magnification, and/or underestimation of asthma symptoms. DESIGN: The study included 270 children with asthma (ages 7-17) and their primary caregivers who completed measures assessing cognitive and psychological factors and a 5 to 6 week symptom perception assessment. MAIN OUTCOME MEASURES: Children's symptom perception scores by use of the Asthma Risk Grid. RESULTS: Children's attentional abilities had more of a bearing on their symptom monitoring abilities than their IQ estimates and psychological symptoms. The more time children took on Trails and Cancellation Tasks and the fewer errors they made on these tasks, the more likely they were to perceive their asthma symptoms accurately. More time on these tasks was associated with more symptom magnification scores, and fewer errors were related to fewer symptom magnification scores. More errors and higher total scores on the Continuous Performance Task were associated with a greater proportion of scores in the danger zone. CONCLUSION: Statistical support was provided for the utility of attentional-based instruments for identifying children who may have problems with perceptual accuracy, and who are at risk for asthma morbidity. PMID- 19290716 TI - Supportive friendships moderate the association between stressful life events and sexual risk taking among African American adolescents. AB - OBJECTIVE: This study examined whether uncontrollable stressful life events were associated with sexual risk taking among adolescents across a 1-year period, and whether supportive friendships modified associations. DESIGN: Participants were 159 sexually active African American adolescents (57% male; mean age [SD] = 17.0 [1.5] years at baseline). Participants were recruited for in-person interviews through random digit dialing in one inner-city neighborhood characterized by high rates of poverty and crime relative to the surrounding city. MAIN OUTCOME MEASURES: Dependent variables included substance use before sexual activity and inconsistent condom use. RESULTS: Among adolescents who reported low levels of supportive friendships, uncontrollable stressors were associated with greater levels of sexual risk taking over time. In contrast, uncontrollable stressors were not associated with sexual risk taking among adolescents who reported high social support from friends; risk taking was typically moderate to high among these adolescents. CONCLUSION: Different processes may explain sexual risk taking among adolescents with varying levels of social support from friends. Adolescents with low support may be prone to engagement in health risk behavior as a stress response, while adolescents with high support may engage in risk behavior primarily due to peer socialization of risk. PMID- 19290717 TI - Health-related quality of life in bereaved HIV-positive adults: relationships between HIV symptoms, grief, social support, and Axis II indication. AB - OBJECTIVE: This study evaluated a model of the impact of borderline and antisocial personality disorder indications on HIV symptoms and health-related quality of life (HRQoL) in AIDS-bereaved adults, accounting for grief severity, social support, and years since HIV diagnosis. DESIGN: Structural equation modeling was used to test the proposed model in a sample of 268 HIV-seropositive adults enrolled in an intervention for coping with AIDS-related bereavement. MAIN OUTCOME MEASURES: Functional assessment of HIV infection, HIV symptoms. RESULTS: The proposed model demonstrated excellent fit with study data and all hypothesized paths were supported. Personality disorder indication was directly related to HIV symptoms and HRQoL and indirectly related through both social support and grief severity. Social support was negatively related to HIV symptoms and positively related to HRQoL, while grief severity was positively related to HIV symptoms and negatively related to HRQoL. Finally, HIV symptoms had a direct negative relationship with HRQoL. CONCLUSION: Personality disorders have a direct negative effect on HIV symptoms and HRQoL and indirect effects through grief severity and social support. PMID- 19290718 TI - Do perceptions of vulnerability and worry mediate the effects of a smoking cessation intervention for women attending for a routine cervical smear test? An experimental study. AB - OBJECTIVE: Numerous correlational studies have examined whether perceptions of vulnerability or worry are better predictors of health-related behavior. The aim of this experimental study was to explore some of the potential causal relationships involved: Are the effects of a brief smoking cessation intervention (for women attending for cervical smear tests) on intention to stop smoking mediated by perceived vulnerability or worry about cervical cancer? DESIGN: A mediation analysis of an experimental study. MAIN OUTCOME MEASURES: Perceived vulnerability to and worry about cervical cancer, and intention to stop smoking in the next month. RESULTS: Questionnaires were completed by 172 (71%) women at 2 week follow-up. Compared with women in the control group, those in the intervention group had higher perceptions of vulnerability, worry, and intention to stop smoking. Personal vulnerability (p < .01) and comparative vulnerability (p < .05) were significant mediators of the relationship between study group and intention to stop smoking. Worry about cervical cancer was not related to intention. CONCLUSION: Worry may be a less important construct in relation to disease prevention behaviors such as smoking cessation. More experimental studies comparing different behaviors are needed to determine the causal relationship between worry and outcomes. PMID- 19290720 TI - The influence of religious participation on the adjustment of female inmates. AB - Incarcerated women at a state correctional facility (N = 213) participated in a study of the relationship between stress, adjustment, institutional misconduct, and degree of personal support derived from religious participation. A series of multivariate analyses of variance investigated differences on adjustment indicators between four groups of inmates who differed on their self-reported support from religious activities, while controlling for self-reported support for other institutional activities. Inmates who received high-level support from participation in religious activities reported significantly less depression, recounted perpetrating fewer aggressive acts, and committed fewer serious institutional infractions than those who did not attend religious activities as well as those who attended but reported receiving low-level support. In addition, inmates reporting a high level of support through their religious activities reported fewer instances of feeling angry, having arguments with inmates and correctional officers, physical fights, and injury than those who reported no participation in religious activities. Results indicate that inmates who perceive that they are receiving personal support from religious activities are better adjusted to the challenges of prison. PMID- 19290721 TI - Beyond point and level systems: moving toward child-centered programming. AB - Many residential treatment facilities and child inpatient units in the United States have been structured by way of motivational programming such as the point and/or level systems. On the surface, they appear to be a straightforward contingency management tool that is based on social learning theory and operant principles. In this article, the authors argue that the assumptions upon which point and level systems are based do not hold up to close empirical scrutiny or theoretical validity, and that point and level system programming is actually counterproductive with some children, and at times can precipitate dangerous clinical situations, such as seclusion and restraint. In this article, the authors critique point and level system programming and assert that continuing such programming is antithetical to individualized, culturally, and developmentally appropriate treatment, and the authors explore the resistance and barriers to changing traditional ways of "doing things." Finally, the authors describe a different approach to providing treatment that is based on a collaborative problem-solving approach and upon which other successful models of treatment have been based. PMID- 19290722 TI - Self-regulation and its relations to adaptive functioning in low income youths. AB - Most studies of self-regulation involving children have linked it to specific outcomes within a single domain of adaptive functioning. The authors examined the association of self-regulation with a range of indices of adaptive functioning among 155 youth ages 8-18 years from families with very low income. Controlling for other explanatory variables, self-regulation was strongly associated with various outcome measures in the areas of mental health, behavior, academic achievement, and social competence. The authors also contrasted youths relatively high and low in self-regulation (the top and bottom quartiles). Youths with good self-regulation had much better indices of adaptive functioning across measures of social competence, academic achievement, grades, problem behaviors, and depression and anxiety than their counterparts with more diminished self regulatory capacities. In addition, youths with better self-regulation skills stated more adaptive responses both in terms of how they coped with past stressful live events and how they would deal with hypothetical stressors. This study indicates that self-regulation is robustly associated with a range of important indices of adaptive functioning across many domains. Findings are discussed in light of their implications for theory and intervention for children of diverse economic backgrounds. PMID- 19290723 TI - Psychiatric disorders in property, violent, and versatile offending detained male adolescents. AB - This study examines the past year prevalence rate of psychiatric disorders in detained male adolescents and the relation between psychiatric disorders and type of offending. The Diagnostic Interview Schedule for Children (DISC-IV) was administered in a sample (N = 245) of male detained adolescents aged 12 to 17 years. Based on lifetime official criminal history, participants were classified into property, violent, and versatile subgroups. High rates of psychiatric disorders were found in all groups. In addition, property offenders reported significantly higher rates of depression, disruptive behavior disorders, substance use disorders and comorbidity than violent and versatile offenders. Overall, versatile offenders did not differ from violent offenders, with the exception of more marijuana use disorder found in violent offenders. This study once more emphasizes that detained boys have substantial mental health needs, a finding that is generalizable across countries. In addition, the current study suggests that classifying detained juveniles by offense subgroups may carry clinical relevance. The long-term impact of these differences, and the possible effects of intervention, should be subject of further research. PMID- 19290724 TI - Paternal and maternal influences on problem behaviors among homeless and runaway youth. AB - Using an Attachment Theory conceptual framework, associations were investigated among positive paternal and maternal relationships, and recent problem behaviors among 501 currently homeless and runaway adolescents (253 males, 248 females). Homeless and runaway youth commonly exhibit problem behaviors such as substance use, various forms of delinquency and risky sex behaviors, and report more emotional distress than typical adolescents. Furthermore, attachments to their families are often strained. In structural equation models, positive paternal relationships significantly predicted less substance use and less criminal behavior, whereas maternal relationships did not have a significant effect on or association with either behavior. Positive maternal relationships predicted less survival sex behavior. Separate gender analyses indicated that among the females, a longer time away from home was significantly associated with a poorer paternal relationship, and more substance use and criminal behavior. Paternal relations, a neglected area of research and often not addressed in attachment theory, should be investigated further. Attachments, particularly to fathers, were protective against many deleterious behaviors. Building on relatively positive relations and attachments may foster family reunifications and beneficial outcomes for at-risk youth. PMID- 19290725 TI - The longitudinal impact of HIV+ parents' drug use on their adolescent children. AB - The impact of parental substance use on the emotional and behavioral adjustment of their adolescent children was examined over 5 years. A representative sample of 220 parents with HIV (PWH) and 330 adolescent children in New York City were repeatedly assessed. Some parents never used marijuana or hard drugs over the 5 years (nonusers). Among those who were users, substance use varied over time. PWH who used substances during a specific 3-month period were classified as active users and those who abstained from substance use were classified as inactive users. Longitudinal regression analyses were used to analyze the impact of variations in patterns of substance use over time on their adolescent children's emotional adjustment and behavioral problems. PWH relapse exacerbated adolescent substance use, trouble with peers, and adolescent emotional distress. Even time limited reductions in parents' substance abuse can have a significant positive impact on their adolescent children's emotional and behavioral adjustment. Interventions that address parental substance use among PWH should be developed to ameliorate the impact of substance use relapse on their adolescents. PMID- 19290726 TI - Cocaine-exposed infant behavior during Still-Face: risk factor analyses. AB - Prenatal cocaine exposure and the role of gender were evaluated using risk factor analyses to determine whether 6-month-old cocaine-exposed male infants demonstrated greater disruptions in infant-caregiver socioemotional interactions during a Still-Face test. Overall, non-cocaine-exposed infants spent more time looking at toys, compared with cocaine-exposed infants; nonexposed female infants spent more time scanning the environment, compared with nonexposed male infants. When caregiver behavior during the Still-Face was evaluated, differences emerged in amount of time the caregiver spent vocalizing to the infant. She vocalized more to a cocaine-exposed infant compared with a nonexposed one; she reduced vocalizing more during the test if the cocaine-exposed infant was female. An exposure by gender interaction emerged in the amount of change in caregiver vocalizations; however, the overarching hypothesis that male cocaine-exposed infants are at higher risk than nonexposed male, nonexposed female, and cocaine exposed female infants was not supported. Because this interaction was evident in this cohort at 24 months, future research is needed to determine at what age an interaction begins to emerge in this cohort. PMID- 19290727 TI - Victimization, anger, and gender: low anger and passive responses work. AB - This study examined the contributions of gender, anger, expectations of positive outcomes, and frequency of victimization by and bullying of peers among school aged children to predict individual differences in intentions to respond to provocative events with nonassertive behavior. Children between the ages of 9 and 13 (N = 505, 246 female, 259 male) completed the Anger Response Inventory, Child Version (Tangney et al., 1996) and measures of victimization and bullying. Results of regression procedures demonstrated that female gender and low anger predicted ignoring and using distraction. Nonassertive responses, low anger, and low victimization predicted expecting more positive outcomes following provocation. Victimization was unrelated to intentions to use nonassertive responses but bullying negatively predicted walking away and using distraction. No modifying effects for gender, victimization, or bullying were found. PMID- 19290728 TI - Non-suicidal self-injury and eating pathology in high school students. AB - Although past research has explored self-injurious behaviors and disordered eating among adults in clinical settings, little research has been conducted examining nonsuicidal self-injury (NSSI) and eating pathology in community samples of adolescents. Four hundred and 40 students were screened for the presence of NSSI; a prevalence rate of 13.9% was found. Those who indicated that they engaged in NSSI (n = 59) and a comparison group of non-self-injurers (n = 57) completed the Eating Disorders Inventory. Results indicate that students who engage in NSSI display significantly more eating pathology than their non-NSSI peers, including poor interoceptive awareness; difficulties with impulse regulation; an increased sense of ineffectiveness, distrust, and social insecurity; and increased bulimic tendencies and body dissatisfaction. Relationships were found between increased lifetime frequency of NSSI behaviors and poor impulse control and deficits in affective regulation. In addition, adolescents who had stopped self-injuring reported comparable rates of eating pathology as did adolescents who continued to self-injure. The theoretical connection between NSSI and eating pathology are discussed with reference to enhancing knowledge regarding the characteristics of NSSI. PMID- 19290729 TI - Female adoptees' perceptions of contact with their birth fathers: satisfactions and dissatisfactions with the process. AB - Qualitative data were analyzed to identify factors associated with adoptees' satisfactions or dissatisfactions in their contact experiences with their birth fathers. Participants were 17 women who had been adopted prior to 2 years of age and had subsequently met their birth fathers. All women completed a questionnaire and 7 also took part in a semistructured interview. Four main themes were identified that affected satisfaction with contact: birth fathers' attributes and behavior, adoptees' attributes (e.g., expectations), behavior of others (e.g., birth mother, birth father's family, and adoptive parents), and circumstances of the conception and relinquishment. The implications of these findings for members of the adoption triangle and those providing postadoption services are discussed. PMID- 19290730 TI - Lesbian and heterosexual preadoptive couples' openness to transracial adoption. AB - This study uses data from 147 White preadoptive couples (54 lesbian, 93 heterosexual) to examine adopters' subjective explanations for why they are open or not open to adopting transracially. Participant perceptions of racial-ethnic diversity in their communities and families, perceptions of family support or nonsupport, and attitudes about race were among the factors they cited as influencing their openness. These findings hold important implications for training and service delivery in transracial adoption. PMID- 19290731 TI - Attributions to discrimination and depression among Latino/as: the mediating role of competence. AB - The present study examined the role of attributions to discrimination and competence in predicting depression among a sample of 93 Latino/a adults. The major findings showed that attributions to discrimination were related to decreases in both general competence and intercultural competence, which were in turn associated with increases in depressive symptoms. This pattern of results suggests that general and intercultural competence partially mediated the relationship between attributions to discrimination and depression. The findings are discussed within the context of the cultural adaptation process and factors that ameliorate Latino/a mental health. In addition, theoretical and practical implications are outlined along with areas of future research. PMID- 19290732 TI - Cultural influences on help-seeking attitudes in Asian American students. AB - There is considerable evidence indicating that Asian American college students have less favorable attitudes toward and are less likely to use mental health services than other ethnic groups in the United States. Because a person's attitudes are often strongly associated with their voluntary behaviors, understanding what influences help-seeking attitudes may help shed light on why Asian American college students refrain from seeking mental health treatment. Andersen's Sociobehavioral Model is commonly used as a guide to understand help seeking in the mainstream population. A modified version of this model that includes culture-related variables (i.e., level of acculturation and stigma tolerance) was used to guide this study. Results indicated that stigma tolerance predicted help-seeking attitudes above and beyond traditional variables associated with help-seeking. These findings suggest that reducing societal stigma and increasing individual tolerance to stigma should be a focus for prevention and intervention programs on college campuses. PMID- 19290733 TI - Cognitive and sensory declines in old age: gauging the evidence for a common cause. AB - Resource accounts of behavioral aging postulate that age-associated impairments within and across intellectual and sensory domains reflect, in part, a common set of senescent alterations in the neurochemistry and neuroanatomy of the aging brain. Hence, these accounts predict sizeable correlations of between-person differences in rates of decline, both within and across intellectual and sensory domains. The authors examined reliability-adjusted variances and covariances in longitudinal change for 8 cognitive measures and for close visual acuity, distant visual acuity, and hearing in 516 participants in the Berlin Aging Study (ages 70 to 103 years at 1st measurement). Up to 6 longitudinal measurements were distributed over up to 13 years. Individual differences in rates of cognitive decline were highly correlated, with a single factor accounting for 60% of the variance in cognitive change. This amount increased to 65% when controlling for age at first measurement, distance to death, and risk of dementia. Contrary to expectations, the correlations between cognitive and sensory declines were only moderate in size, underscoring the need to delineate both domain-general and function-specific mechanisms of behavioral senescence. PMID- 19290734 TI - The association between age and the frequency of nouns selected for production. AB - This article examines whether age-related difficulties in word retrieval are reflected in the lexical frequency of the nouns selected for production. Participants (N = 136; age range 20-85 years) completed a picture-naming test, a semantic fluency task, and a picture description task. Each noun in the speech samples was examined for its lexical frequency according to published word counts and was rated subjectively by 40 young and 40 older adults. In addition, the cumulative occurrence of each noun was calculated across all descriptions. Cumulative analysis was conducted on fluency protocols as well. Results show that age was associated with a decrease in total scores on structured single-word production tests as well as a decrease in the lexical frequency of nouns selected for production. Age was also associated with the production of words that appeared in 1 picture description only or in 1 fluency protocol only. Instead of selecting the most common nouns and thus avoiding age-associated retrieval difficulties, older speakers appear to produce more infrequent nouns, most likely because they have a larger vocabulary from which to choose these nouns. PMID- 19290735 TI - Adult aging effects on semantic and episodic priming in word recognition. AB - Two experiments compared automatic semantic and episodic priming effects in adult aging. In the 1st experiment, target words were semantically primed; in the 2nd experiment, targets were primed by repetition of semantically unrelated words. Both experiments involved a pronunciation task with response signals at fixed times following target onset. Consequently, priming was measured as improvement in the percentage of correct responses. Priming was also calculated with speed accuracy measures of intercept and slope. Both types of priming effect were significant in the percentage correct and slope measures, but no age group differences were found. Furthermore, the magnitudes of the priming effects were equivalent. The age-resistant nature of semantic and episodic priming, as well as evidence for a common theoretical mechanism, is discussed. PMID- 19290736 TI - Smarter in middle age, faster in old age: a cross-lagged panel analysis of reaction time and cognitive ability over 13 years in the West of Scotland Twenty 07 Study. AB - Participants in the West of Scotland Twenty-07 Study performed reaction time tasks and took the Alice Heim 4 Part 1 test (AH4) of intelligence twice, 13 years apart. Cross-lagged associations between speed of processing and AH4 were examined using latent variables in structural equation modeling. The stability coefficients of the latent traits of processing speed and of AH4 score across 13 years were .49 and .89, respectively. There was a significant association (-.21) between AH4 score at age 56 and speed of processing at age 69 but not vice versa. The results fail to support the theory that processing speed is a foundation for successful cognitive aging but support a hypothesis that suggests that higher general intelligence might be associated with lifestyle and other factors that preserve processing speed. PMID- 19290737 TI - Genetic and environmental transactions linking cognitive ability, physical fitness, and education in late life. AB - Cognitive ability and physical fitness are important to the ability to live independently in late life. Both are also related to level of attained education, with better educated older adults tending to display better cognitive ability and better late-life physical health. Chronic illnesses that affect both physical and cognitive function, lifetime cognitive ability that facilitates healthy lifestyle choices, and general biological aging processes have been offered as 3 explanations for the late-life physical-cognitive correlation. Education is generally assumed to provide a protective environment. The authors used a sample of 1,053 twin pairs aged 70 and over and gene-environment moderation models to explore 5 hypotheses that could help to disentangle the genetic and environmental transactions involving physical and cognitive functions and education. Results provide some support for all 3 explanations for the physical-cognitive correlation and indicate the ways in which better education may support better function and lack of education may undermine it. PMID- 19290738 TI - Age differences in the effects of domain knowledge on reading efficiency. AB - In the present study, the author investigated age differences in the effects of knowledge on the efficiency with which information is processed while reading. Individuals between 18 and 85 years of age, with varying levels of cooking knowledge, read and recalled a series of short passages within the domain of cooking. Reading efficiency was operationalized as time spent reading divided by the amount recalled for each passage. Results showed that reading efficiency increased with increasing levels of knowledge among older but not younger adults. Similarly, those with smaller working memory capacities showed increasing efficiency with increasing knowledge. These findings suggest that knowledge promotes a more efficient allocation policy that is particularly helpful in later life, perhaps due to age-related declines in working memory capacity. PMID- 19290739 TI - Interacting effects of cognitive load and adult age on the regularity of whole body motion during treadmill walking. AB - We investigated effects of concurrent cognitive task difficulty (n-back) on the regularity of whole-body movements during treadmill walking in women and men from 3 age groups (20-30, 60-70, and 70-80 years old). Using principal component analysis of individual gait patterns, we separated main (regular) from residual (irregular) components of whole-body motion. Proportion of residual variance (RV) was used as an index of gait irregularity. The gait in all age groups became more regular (reduced RV) upon introduction of a simple cognitive task (1-back), relative to walking without a concurrent cognitive task. In contrast, parametrically increasing working memory load from 1-back to 4-back led to age differential effects, with gait patterns becoming more regular in those 20-30 years old, becoming less regular in those 70-80 years old, and showing no significant effects in those 60-70 years old. Our results support the dual process account of sensorimotor-cognitive interactions (O. Huxhold, S.-C. Li, F. Schmiedek, and U. Lindenberger, 2006), with age-general effects of internal versus external attentional focus and age-specific effects of resource competition with increasing cognitive task difficulty. PMID- 19290740 TI - Does differential strategy use account for age-related deficits in working-memory performance? AB - The strategy-deficit hypothesis states that age differences in the use of effective strategies contribute to age-related deficits in working memory span performance. To evaluate this hypothesis, strategy use was measured with set-by set strategy reports during the Reading Span task (Experiments 1 and 2) and the Operation Span task (Experiment 2). Individual differences in the reported use of effective strategies accounted for substantial variance in span performance. In contrast to the strategy-deficit hypothesis, however, young and older adults reported using the same proportion of normatively effective strategies on both span tasks. Measures of processing speed accounted for a substantial proportion of the age-related variance in span performance. Thus, although use of normatively effective strategies accounts for individual differences in span performance, age differences in effective strategy use cannot explain the age related variance in that performance. PMID- 19290741 TI - Priming of familiar and unfamiliar visual objects over delays in young and older adults. AB - Although priming of familiar stimuli is usually age invariant, little is known about how aging affects priming of preexperimentally unfamiliar stimuli. Therefore, this study investigated the effects of aging and encoding-to-test delays (0 min, 20 min, 90 min, and 1 week) on priming of unfamiliar objects in block-based priming paradigms. During the encoding phase, participants viewed pictures of novel objects (Experiments 1 and 2) or novel and familiar objects (Experiment 3) and judged their left-right orientation. In the test block, priming was measured using the possible-impossible object-decision test (Experiment 1), symmetric-asymmetric object-decision test (Experiment 2), and real-nonreal object-decision test (Experiment 3). In Experiments 1 and 2, young adults showed priming for unfamiliar objects at all delays, whereas older adults whose baseline task performance was similar to that of young adults did not show any priming. Experiment 3 found no effects of age or delay on priming of familiar objects; however, priming of unfamiliar objects was only observed in the young participants. This suggests that when older adults cannot rely on preexisting memory representations, age-related deficits in priming can emerge. PMID- 19290742 TI - No age differences in complex memory search: older adults search as efficiently as younger adults. AB - In 2 experiments, the authors investigated age differences in memory search under 4 conditions: forward search, backward search, random search, and fixed irregular search. Both search slopes and serial position curves were investigated. Mixing conditions led to smaller age differences than blocking conditions, suggesting that younger adults have an advantage over older adults when strategies can be applied to memory scanning. All age differences in scanning rates, however, disappeared when age differences in a magnitude-judgment control task were controlled for, showing that age differences in memory scanning tasks are not because of the scanning process per se, but because of attention, sensorimotor speed, and decision processes. In both experiments, the serial position curves of older adults echoed those of younger adults closely, demonstrating that younger and older adults use the same scanning processes. PMID- 19290743 TI - Age and education effects on relationships of cognitive test scores with brain structure in demographically diverse older persons. AB - This study examined how age and education influence the relationship between neuropsychological test scores and brain structure in demographically diverse older adults spanning the range from normal cognition to dementia. A sample of 351 African Americans, 410 Hispanics, and 458 Whites underwent neuropsychological testing. Volumetric magnetic resonance imaging (MRI) measures of total brain, white matter hyperintensity, and hippocampus were available for 79 African Americans, 102 Hispanics, and 134 Whites. The authors used latent variable modeling to examine effects of age, education, and brain volumes on test scores and determine how much variance brain volumes explained in unadjusted and age- and education-adjusted scores. Age adjustment resulted in weaker relationships of test scores with MRI variables; adjustment for ethnicity yielded stronger relationships. Education adjustment increased relationships with MRI variables in the combined sample and Hispanics, made no difference in Whites, but decreased some associations in African Americans. Results suggest that demographic adjustment is beneficial when demographic variables are strongly related to test scores independent of measures of brain structure, but adjustment has negative consequences when effects of demographic characteristics are mediated by brain structure. PMID- 19290744 TI - Cognitive ability at age 11 and 70 years, information processing speed, and APOE variation: the Lothian Birth Cohort 1936 study. AB - The e4 allele of the apolipoprotein E (APOE) gene confers risk of Alzheimer's disease and, in some studies, relates to cognitive ability and decline in older people without Alzheimer's disease. Its relationship with processing speed, a contributor to cognitive decline with age, is largely unknown. This study tests the association of APOE with cognition and speed, with and without covarying childhood mental ability. The 1,013 participants were tested on cognitive ability at age 11 as part of the Scottish Mental Survey of 1947 and, at age 70, were tested on reasoning, working memory, information processing speed, and executive function. The results showed that APOE was associated with the general cognitive factor, 2 nonverbal tests, and choice reaction time (RT) variability; as expected, the e4 allele was the risk allele. RT measures and a general speed factor were nonlinearly related to APOE when factoring childhood ability (p < .05): The correlation between childhood ability and speed was lower in e4 allele carriers. APOE has an influence on nonverbal cognition in old age and interacts with childhood IQ to influence processing speed. PMID- 19290745 TI - Neural basis for recognition confidence in younger and older adults. AB - Although several studies have examined the neural basis for age-related changes in objective memory performance, less is known about how the process of memory monitoring changes with aging. The authors used functional magnetic resonance imaging to examine retrospective confidence in memory performance in aging. During low confidence, both younger and older adults showed behavioral evidence that they were guessing during recognition and that they were aware they were guessing when making confidence judgments. Similarly, both younger and older adults showed increased neural activity during low- compared to high-confidence responses in the lateral prefrontal cortex, anterior cingulate cortex, and left intraparietal sulcus. In contrast, older adults showed more high-confidence errors than younger adults. Younger adults showed greater activity for high compared to low confidence in medial temporal lobe structures, but older adults did not show this pattern. Taken together, these findings may suggest that impairments in the confidence-accuracy relationship for memory in older adults, which are often driven by high-confidence errors, may be primarily related to altered neural signals associated with greater activity for high-confidence responses. PMID- 19290746 TI - Aging, vascular risk, and cognition: blood glucose, pulse pressure, and cognitive performance in healthy adults. AB - Advanced age is associated with decline in many areas of cognition as well as increased frequency of vascular disease. Well-described risk factors for vascular disease, such as diabetes and arterial hypertension, have been linked to cognitive deficits beyond those associated with aging. To examine whether vascular health indices such as fasting blood glucose levels and arterial pulse pressure can predict subtle deficits in age-sensitive abilities, the authors studied 104 healthy adults (ages 18 to 78) without diagnoses of diabetes or hypertension. Whereas results revealed a classic pattern of age-related differences in cognition, preprandial blood glucose level and pulse pressure independently and differentially affected cognitive performance. High-normal blood glucose levels were associated with decreased delayed associative memory, reduced accuracy of working memory processing among women, and slower working memory processing among men. Elevated pulse pressure was associated with slower perceptual-motor processing. Results suggest that blood glucose levels and pulse pressure may be sensitive indicators of cognitive status in healthy adults; however, longitudinal research is needed to determine whether such relatively mild elevations in this select group predict age-related cognitive declines. PMID- 19290747 TI - Repelling the young and attracting the old: examining age-related differences in saccade trajectory deviations. AB - In the present study, the authors examined age-related differences in saccade curvature as older and younger adults looked to an X target that appeared concurrently with an O distractor. They used a fixation gap procedure to introduce variance into the saccadic latencies of both groups. Consistent with earlier findings, younger adults' early onset saccades curved toward the distractor (as the distractor competed with the target for response selection), while late-onset saccades curved away from the distractor (as the distractor location became inhibited over time). In contrast, older adults' saccades gradually decreased in curvature toward the distractor, but at no point along the latency continuum did they show deviations away. These results suggest that while the local inhibitory mechanisms responsible for decreases in curvature toward distractors may be preserved with age, aging may lead to a selective decline in the frontal inhibitory mechanisms responsible for deviations away from distractors. PMID- 19290748 TI - Expectations about memory change across the life span are impacted by aging stereotypes. AB - This study examined whether expectations about memory change with age vary for different personality types. Four adjectives from each of M. L. Hummert, T. A. Garstka, J. L. Shaner, and S. Strahm's (1994) age-stereotype trait sets were selected to create 11 adjective clusters varying in both valence (positive vs. negative) and relevance to memory functioning. There were 373 participants in 3 age groups who rated the memory abilities of target adults, defined by the adjective clusters, across the adult life span. Consistent with past studies, participants believed in age-related memory decline. However, participants rated target adults with positive personality traits as having better memory ability and less age-related memory decline than target adults with negative personality traits. This effect was larger when the traits were relevant to memory than when they were not. Finally, older participants were more strongly influenced by both the valence and the relevance of the personality descriptions than younger participants. PMID- 19290749 TI - Perceived income inadequacy as a predictor of psychological distress in Alzheimer's caregivers. AB - The authors examined perceived income inadequacy as a predictor of self-reported depressive symptomatology and anxiety in the original sites of the Resources for Enhancing Alzheimer's Caregiver Health I project. Perceived income inadequacy, self-reported household income, and control factors (e.g., subjective health) were entered into hierarchical regression analyses predicting psychological distress. Findings suggest that perceived income inadequacy and not household income significantly predicted more self-reported depressive symptomatology and greater self-reported anxiety. This supports previous findings that objective income measures alone are not adequate indicators of socioeconomic status in older adults. PMID- 19290750 TI - Prospective associations between marital discord and depressive symptoms in middle-aged and older adults. AB - Data from both spouses in a population-based sample of middle-aged and older adults (N = 1,869 couples) were used to evaluate the 2-year prospective association between marital discord and depressive symptoms. Nested path analyses indicated that (a) baseline marital discord predicted one's own depressive symptoms at follow-up, (b) baseline depressive symptoms predicted one's own marital discord at follow-up, (c) baseline depressive symptoms predicted partners' marital discord at follow-up, and (d) there were no gender differences in the magnitudes of the pathways between one's own baseline depressive symptoms and one's own marital discord at follow-up or between one's own baseline marital discord and one's own depressive symptoms at follow-up. These results suggest a bidirectional longitudinal association between marital discord and depressive symptoms in middle-aged and older adults. PMID- 19290751 TI - Outcomes from the Resources for Enhancing Alzheimer's Caregiver Health (REACH) program for bereaved caregivers. AB - Although preventive efforts for bereaved individuals have not been shown to be particularly effective in past studies, it has been suggested that intervention effects might be underestimated due to methodological issues such as short follow up assessments. Thus, the present study aimed to assess the efficacy (as whole intervention packages and as component parts) of the Resources for Enhancing Alzheimer's Caregiver Health (REACH) interventions in preventing complicated grief, normal grief, and depressive symptoms at a longer term follow-up assessment among bereaved caregivers (N = 224). On average, active interventions showed a statistically significant effect on normal grief symptoms (d = 0.28), exhibited a trend toward improvement on complicated grief symptoms (d = 0.25), and demonstrated little impact on depressive symptoms (d = 0.09). Interestingly, the examination of intervention components revealed differential effects, whereby cognitive and behavioral strategies were most effective at reducing levels of complicated grief, information and emotional support were most effective for addressing normal grief, and environmental modifications were most effective for ameliorating depressive symptoms. These findings suggest that different preloss interventions might be warranted depending on a caregiver's unique clinical presentation and combination of risk factors. PMID- 19290752 TI - Interference and facilitation in spatial working memory: age-associated differences in lure effects in the n-back paradigm. AB - Working memory (WM) declines prominently during normal aging. The mechanisms underlying this decline are not fully understood. The authors analyzed performance on 2 versions of a 2-back spatial WM task to assess younger and older adults' responses to lures (i.e., nontarget items that match an item earlier in the sequence but not at the current target lag). Results demonstrate lure interference effects that are particularly pronounced among older adults. At the same time, however, older adults showed facilitation for targets. Taken together, these findings suggest that the contribution of familiarity signals to WM performance increases during normal aging. PMID- 19290753 TI - Illusory recollection in older adults and younger adults under divided attention. AB - The authors investigated the effect of divided attention, study-list repetition, and age on recollection and familiarity. Older and younger adults under full attention and younger adults under divided attention at study viewed word lists highly associated with a single unstudied word (critical lure) once or three times, and subsequently performed a remember-know recognition test. Younger adults made fewer false remember responses to critical lures from repeated study lists, whereas younger adults under divided attention and older adults both showed an increase with repetition. Findings suggest older adults' susceptibility to illusory memories is related to a deficit in available attention during encoding. PMID- 19290754 TI - Effects of regulating emotions on cognitive performance: what is costly for young adults is not so costly for older adults. AB - The authors examined whether instructions to regulate emotions after a disgust inducing film clip created an equally costly cognitive load across adulthood. Young and older adults across all instructional conditions initially demonstrated increased working memory performance after mood induction, typical of practice effects. Age-group differences emerged at the 2nd postinduction trial. When instructed to down-regulate disgust feelings, older adults' performance continually increased, whereas young adults' performance dropped. Instructions to maintain disgust did not affect working memory performance. Consistent with claims that older adults are more effective at regulating emotions, findings indicate that intentional down-regulation of negative emotions may be less costly in older age. PMID- 19290755 TI - Electromyographic evidence for age-related differences in the mimicry of anger. AB - Although older adults have difficulty recognizing all facial emotions, they have particular difficulty decoding expressions of anger. Since disruption of facial mimicry impairs emotion recognition, electromyography of the corrugator supercilii (i.e., brow) muscle region was used to test whether there are age differences in anger mimicry. Associations between mimicry and emotion recognition were also assessed. The results indicated that although there were no age differences in anger mimicry, older (but not young) adults' corrugator responses to angry expressions were associated with reduced anger recognition. Implications for understanding emotion recognition difficulties in older adulthood are discussed. PMID- 19290757 TI - The stereotype-matching effect: greater influence on functioning when age stereotypes correspond to outcomes. AB - Older individuals assimilate, and are targeted by, contradictory positive and negative age stereotypes. It was unknown whether the influence of stereotype valence is stronger when the stereotype content corresponds to the outcome domain. We randomly assigned older individuals to either positive-cognitive, negative-cognitive, positive-physical, or negative-physical subliminal-age stereotype groups and assessed cognitive and physical outcomes. As predicted, when the age stereotypes corresponded to the outcome domains, their valence had a significantly greater impact on cognitive and physical performance. This suggests that if a match occurs, it is more likely to generate expectations that become self-fulfilling prophecies. PMID- 19290758 TI - The realism in older people's confidence judgments of answers to general knowledge questions. AB - The study investigated 2 aspects of the accuracy (i.e., realism) of confidence judgments of persons age 60-93 years (N = 1,384) regarding their answers to general knowledge questions. These aspects are the level of confidence (calibration) in relation to the proportion of correct answers and the ability to discriminate between correct and incorrect answers by means of confidence judgments. No age differences were found for either of the 2 aspects. Gender differences were found for proportion of correct answers and confidence but not for the realism in the confidence judgments. PMID- 19290759 TI - Social inappropriateness, executive control, and aging. AB - Age-related deficits in executive control might lead to socially inappropriate behavior if they compromise the ability to withhold inappropriate responses. Consistent with this possibility, older adults in the current study showed greater social inappropriateness than younger adults--as rated by their peers- and this effect was mediated by deficits in executive control as well as deficits in general cognitive ability. Older adults also responded with greater social inappropriateness to a provocative event in the laboratory, but this effect was unrelated to executive functioning or general cognitive ability. These findings suggest that changes in both social and cognitive factors are important in understanding age-related changes in social behavior. PMID- 19290760 TI - Understanding the influence of demographic and psychological variables on retirement planning. AB - The authors examined the degree to which workers were engaging in financial, health, interpersonal/leisure, and work planning for retirement, exploring whether demographic and psychological variables inhibited or promoted planning in each of these domains. Planning in each domain was influenced by a unique set of variables. Goals emerged as a consistent and positive predictor of planning. Gender accounted for health and interpersonal/leisure planning, while work planning behavior was negatively predicted by income. Time perspective also helped to clarify the amount of retirement planning undertaken in the financial and interpersonal/leisure domains. Practical implications for designing retirement interventions are discussed. PMID- 19290761 TI - Emotion experience, expression, and regulation in Alzheimer's disease. AB - Alzheimer's disease (AD) is associated with intact experience but abnormal expression of emotion. Because emotion regulation is important in determining levels of experienced and expressed emotion, individuals with AD and control participants were asked to watch film clips under conditions of spontaneous expression, suppression, or amplification of emotion. Both groups had difficulties with behavioral amplification that were related to performance on a measure of theory of mind. However, intentional use of suppression was intact even for those with AD, consistent with models of aging that regard some emotion control processes as being relatively more automatic in older adulthood. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290762 TI - The accuracy of recidivism risk assessments for sexual offenders: a meta-analysis of 118 prediction studies. AB - This review compared the accuracy of various approaches to the prediction of recidivism among sexual offenders. On the basis of a meta-analysis of 536 findings drawn from 118 distinct samples (45,398 sexual offenders, 16 countries), empirically derived actuarial measures were more accurate than unstructured professional judgment for all outcomes (sexual, violent, or any recidivism). The accuracy of structured professional judgment was intermediate between the accuracy found for the actuarial measures and for unstructured professional judgment. The effect sizes for the actuarial measures were moderate to large by conventional standards (average d values of 0.67-0.97); however, the utility of the actuarial measures will vary according to the referral question and samples assessed. Further research should identify the psychologically meaningfully factors that contribute to risk for reoffending. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290763 TI - Some evidence for multidimensional biculturalism: confirmatory factor analysis and measurement invariance analysis on the Bicultural Involvement Questionnaire Short Version. AB - In the present study, the authors empirically investigated a multidimensional conceptualization of acculturation using the Bicultural Involvement Questionnaire Short Version (BIQ-S; J. Szapocznik, W. M. Kurtines, & T. Fernandez, 1980). Baseline data from four prevention trials, representing a total of 893 adolescent and 880 guardian participants of Hispanic descent, were used to examine the factor structure of BIQ-S scores. Results of exploratory and confirmatory factor analyses supported a four-factor solution. The factors were labeled as follows: (a) Comfort With Use of Spanish Language, (b) Comfort With Use of English Language, (c) Enjoyment of Hispanic Cultural Activities, and (d) Enjoyment of American Cultural Activities. The findings support an expansion of the theoretical conceptualization of biculturalism and suggest that within both the heritage and the receiving cultures, comfort with language and enjoyment of other cultural components represent different factors. Measurement invariance analyses revealed stability of the factor structure, as well as some measurement differences, between adolescents and their guardians. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290764 TI - Development and evaluation of the Balanced Index of Psychological Mindedness (BIPM). AB - Psychological mindedness (PM) refers to a person's interest and ability to be in touch with and reflect on his or her psychological states and processes. In this multipart study, the authors present the Balanced Index of Psychological Mindedness (BIPM). The psychometric properties and clinical relevance of this 14 item self-report scale were examined in 2 community samples and a mental health patient sample in the Netherlands (total N = 1,211). The 7-item Interest and Insight subscales of the BIPM showed good internal consistency (Cronbach's alphas = .85 and .76, respectively), test-retest reliability (r = .63 and .71, respectively), and construct validity (e.g., r > .40 with related constructs). PM was higher in women than in men. Relatively low PM was found among poorly educated people and in mental health patients (ps < .001). Insight, but not Interest, correlated negatively with symptoms of psychological distress (r = -.21 to -.36, p < .05) for 6 out of 8 symptom scales in mental health patients. These data suggest that the BIPM is a concise, valid, and clinically relevant self report scale that may enhance research on PM in clinical groups as well as the general population. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290765 TI - Factor structure of the Hare Psychopathy Checklist: youth version in German female and male detainees and community adolescents. AB - Substantial evidence exists for 3- and 4-factor models of psychopathy underlying patterns of covariation among the items of the Psychopathy Checklist-Revised (PCL R) in diverse adult samples. Although initial studies conducted with the Psychopathy Checklist: Youth Version (PCL:YV) indicated reasonable fit for these models in incarcerated male adolescents in the United States and the United Kingdom, only one published study has addressed the factor structure of PCL:YV psychopathy in female adolescents, and no prior studies have addressed it outside of these countries. We used confirmatory factor analysis to investigate the factor structure underlying PCL:YV scores in 314 incarcerated (143 male, 171 female) and 193 in-school (99 male, 94 female) adolescents, ages 14 to 19 years. The 2-factor model provided adequate fit only for incarcerated male adolescents and the 4-factor model was problematic in all samples, but the 3-factor solution provided an adequate model in incarcerated and community male adolescents. None of the models provided consistently acceptable fit among female adolescents. Current findings provide evidence for the robustness of the 3-factor model of psychopathy in incarcerated and community male adolescent samples but raise doubts about the applicability of this model to female adolescents. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290766 TI - An exploration of the base rate scores of the Millon Clinical Multiaxial Inventory-III. AB - The Millon Clinical Multiaxial Inventory (3rd ed.; MCMI-III) is a widely used psychological assessment of clinical and personality disorders. Unlike typical tests, the MCMI-III uses a base-rate score transformation to incorporate prior probabilities of disorder (i.e., base rates) in test output and diagnostic thresholds. The authors describe the base rate transformation and contend that its supporting documentation in the MCMI-III manual is incomplete and fails to meet interdisciplinary test documentation standards. They show that the MCMI III's base rate transformation is not optimal, and they derive an optimal alternative transformation using Bayes' theorem. Bayes transformation formulae for 7 exponential family distributions are given. The authors discuss the effect of the base rate transformation and further argue that the MCMI-III's use of a single base rate per diagnostic category is ill-advised. They argue that base rates differ among clinical settings and that tests like the MCMI-III should flexibly incorporate a base rate of disorder pertinent to the examinee's characteristics, such as demographics, chief complaint, clinical history, or other variables. They explain how this can readily be accomplished. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290767 TI - Detecting psychopathy from thin slices of behavior. AB - This study is the first to demonstrate that features of psychopathy can be reliably and validly detected by lay raters from "thin slices" (i.e., small samples) of behavior. Brief excerpts (5 s, 10 s, and 20 s) from interviews with 96 maximum-security inmates were presented in video or audio form or in both modalities combined. Forty raters used these excerpts to complete assessments of overall psychopathy and its Factor 1 and Factor 2 components, various personality disorders, violence proneness, and attractiveness. Thin-slice ratings of psychopathy correlated moderately and significantly with psychopathy criterion measures, especially those related to interpersonal features of psychopathy, particularly in the 5- and 10-s excerpt conditions and in the video and combined channel conditions. These findings demonstrate that first impressions of psychopathy and related constructs, particularly those pertaining to interpersonal functioning, can be reasonably reliable and valid. They also raise intriguing questions regarding how individuals form first impressions and about the extent to which first impressions may influence the assessment of personality disorders. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290768 TI - Evaluating the validity of computerized content analysis programs for identification of emotional expression in cancer narratives. AB - Psychological interventions provide linguistic data that are particularly useful for testing mechanisms of action and improving intervention methodologies. For this study, emotional expression in an Internet-based intervention for women with breast cancer (n = 63) was analyzed via rater coding and 2 computerized coding methods (Linguistic Inquiry and Word Count [LIWC] and Psychiatric Content Analysis and Diagnosis [PCAD]). Although the computerized coding methods captured most of the emotion identified by raters (LIWC sensitivity = .88; PCAD sensitivity = .83), both over-identified emotional expression (LIWC positive predictive value = .31; PCAD positive predictive value = .19). Correlational analyses suggested better convergent and discriminant validity for LIWC. The results highlight previously unrecognized deficiencies in commonly used computerized content-analysis programs and suggest potential modifications to both programs that could improve overall accuracy of automated identification of emotional expression. Although the authors recognize these limitations, they conclude that LIWC is superior to PCAD for rapid identification of emotional expression in text. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290769 TI - Interpersonal characteristics of male criminal offenders: personality, psychopathological, and behavioral correlates. AB - Interest in conceptualizing the interpersonal style of individuals who engage in serious antisocial behavior has increased in recent years. This study examines the personality, psychopathological, and behavioral correlates of interpersonal dominance and warmth, as operationalized via scales of the Personality Assessment Inventory (L. Morey, 2007), across several samples of male prison inmates (combined N = 1,062). Consistent with theory, multivariate analyses indicated that low warmth and, to a lesser extent, high dominance were associated with antisocial and paranoid traits, specifically, and externalizing-spectrum psychopathology more generally, whereas borderline traits and internalizing spectrum psychopathology were uniquely associated with low interpersonal warmth. Among smaller subsamples of inmates followed prospectively, high dominance and, to a lesser extent, low warmth predicted general and aggressive institutional misconduct, whereas dominance uniquely predicted staff ratings of treatment noncompliance/failure. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290770 TI - To sum or not to sum: taxometric analysis with ordered categorical assessment items. AB - Meehl's taxometric method has been shown to differentiate between categorical and dimensional data, but there are many ways to implement taxometric procedures. When analyzing the ordered categorical data typically provided by assessment instruments, summing items to form input indicators has been a popular practice for more than 20 years. A Monte Carlo study compared the accuracy of taxometric analyses implemented in the traditional way (without summing items) and taxometric analyses implemented with the summed-input method. These analyses generated no support for the summed-input method, which substantially reduced discriminating power for 2 of the 3 procedures studied. Accuracy was highest when 5 or more indicators and 4 or more ordered categories were used. Findings from the simulation study were then used to help interpret the results for taxometric analyses of antisocial personality disorder criteria with real research data. In this example, the traditional method yielded clearer results than the summed input method. Implications for the use and further study of the taxometric method in assessment research are discussed. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290771 TI - Detection of overreporting of psychopathology on the Personality Assessment Inventory: a meta-analytic review. AB - The Personality Assessment Inventory (L. C. Morey, 1991) includes 3 measures for identifying overreporting of psychopathology: the Negative Impression scale (NIM), Malingering Index (MAL), and Rogers Discriminant Function (RDF). Meta analysis revealed that each measure was a strong predictor of uncoached (NIM, d = 1.48, k = 23; MAL, d = 1.15, k = 19; RDF, d = 1.13, k = 15) and coached malingering (NIM, d = 1.59, k = 8; MAL, d = 1.00, k = 6; RDF, d = 1.65, k = 3). For uncoached malingering, effects were larger in simulation than criterion groups studies, for identifying feigning of severe mental disorders than mood/anxiety disorders, and when feigners were compared to unimpaired honest respondents as opposed to patients. Cut scores of NIM > or = 81 and MAL > or = 3 resulted in the highest overall classification rates for identifying feigning. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290772 TI - Assessment of borderline personality features in population samples: is the Personality Assessment Inventory-Borderline Features scale measurement invariant across sex and age? AB - Borderline personality disorder (BPD) is more often diagnosed in women than in men, and symptoms tend to decline with age. Using a large community sample, the authors investigated whether sex and age differences in four main features of BPD, measured with the Personality Assessment Inventory-Borderline Features scale (PAI-BOR; Morey, 1991), are a result of measurement bias or if they represent true differences. The PAI-BOR was completed by four Sex x Age groups (N = 6,838). Multigroup confirmatory factor analysis showed that the PAI-BOR is measurement invariant across sex and age. Compared with men, women reported more borderline characteristics for affective instability, identity problems, and negative relationships but not for self-harm. Younger men had higher scores for identity problems and self-harm than did older men. Younger women had higher scores for identity problems and affective instability than did older women. Results suggest that the PAI-BOR can be used to study the etiology of BPD features in population based samples and to screen for BPD features in clinical settings in both men and women of varying ages. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290773 TI - A cross-cultural study of the psychometric properties of the Reactive-Proactive Aggression Questionnaire among Italian nonclinical adolescents. AB - Five independent studies were used to test the hypothesis that a reliable 2 factor structure underlies the Reactive-Proactive Aggression Questionnaire (RPQ) items and that the 2 scales show distinct patterns of association with personality and bullying behavior measures. Study 1 (N = 1,447) gave evidence of a clear 2-factor structure of RPQ items with factor loading matrices closely matching reactive (congruence coefficient = .90) and proactive (congruence coefficient = .91) models of item assignment. The RPQ 2-factor structure was consistently replicated in Study 2 (N = 662), as well as across the remaining 3 studies. In Study 3 (N = 536), Neuroticism differentiated reactive and proactive forms of aggression. In Study 4 (N = 674), self-reports of bullying behaviors were selectively correlated with proactive aggression. Findings confirm and extend the differential correlates of proactive-reactive aggression and also support the psychometric properties of the RPQ in a different cultural context. Finally, in Study 5 (N = 347), the RPQ scales showed adequate 2-month test-retest reliability. (PsycINFO Database Record (c) 2009 APA, all rights reserved). PMID- 19290775 TI - An exploratory survey on the views of European tissue engineers concerning the ethical issues of tissue engineering research. AB - We present the first exploratory survey about the views of tissue engineers on the ethical issues of tissue engineering (TE), conducted among participants of a large European TE consortium. We analyzed the topics for which ethical guidance is necessary and the preferred dissemination channels, which are relevant issues and goals of clinical trials with human tissue-engineered products, and which information is to be given to trial participants. The need for comprehensive, specific ethical guidance of TE is a first key finding of this survey. Second, it becomes clear that little clarity exists on some crucial issues in the setup and conduct of clinical trials in TE. Identifying the unique features of TE and their repercussions for the ethical conduct of TE research and therapy is necessary. Third, prospective trial participants are to be informed about a wide variety of issues before taking part in the trial. PMID- 19290776 TI - Absence of transverse tubules contributes to non-uniform Ca(2+) wavefronts in mouse and human embryonic stem cell-derived cardiomyocytes. AB - Mouse (m) and human embryonic stem cell-derived cardiomyocytes (hESC-CMs) are known to exhibit immature Ca(2+) dynamics such as small whole-cell peak amplitude and slower kinetics relative to those of adult. In this study, we examined the maturity and efficiency of Ca(2+)-induced Ca(2+) release in m and hESC-CMs, the presence of transverse (t) tubules and its effects on the regional Ca(2+) dynamics. In m and hESC-CMs, fluorescent staining and atomic force microscopy (AFM) were used to detect the presence of t-tubules, caveolin-3, amphiphysin-2 and colocalization of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs). To avoid ambiguities, regional electrically-stimulated Ca(2+) dynamics of single ESC-CMs, rather than spontaneously beating clusters, were measured using confocal microscopy. m and hESC-CMs showed absence of dyads, with neither t tubules nor colocalization of DHPRs and RyRs. Caveolin-3 and amphiphysin-2, crucial for the biogenesis of t-tubules with robust expression in adult CMs, were also absent. Single m and hESC-CMs displayed non-uniform Ca(2+) dynamics across the cell that is typical of CMs deficient of t-tubules. Local Ca(2+) transients exhibited greater peak amplitude at the peripheral than at the central region for m (3.50 +/- 0.42 vs. 3.05 +/- 0.38) and hESC-CMs (2.96 +/- 0.25 vs. 2.72 +/- 0.25). Kinetically, both the rates of rise to peak amplitude and transient decay were faster for the peripheral relative to the central region. Immature m and hESC-CMs display unsynchronized Ca(2+) transients due to the absence of t-tubules and gene products crucial for their biogenesis. Our results provide insights for driving the maturation of ESC-CMs. PMID- 19290777 TI - A disruption in iron-sulfur center biogenesis via inhibition of mitochondrial dithiol glutaredoxin 2 may contribute to mitochondrial and cellular iron dysregulation in mammalian glutathione-depleted dopaminergic cells: implications for Parkinson's disease. AB - Parkinson's disease (PD) is characterized by early glutathione depletion in the substantia nigra (SN). Among its various functions in the cell, glutathione acts as a substrate for the mitochondrial enzyme glutaredoxin 2 (Grx2). Grx2 is involved in glutathionylation of protein cysteine sulfhydryl residues in the mitochondria. Although monothiol glutathione-dependent oxidoreductases (Grxs) have previously been demonstrated to be involved in iron-sulfur (Fe-S) center biogenesis, including that in yeast, here we report data suggesting the involvement of mitochondrial Grx2, a dithiol Grx, in iron-sulfur biogenesis in a mammalian dopaminergic cell line. Given that mitochondrial dysfunction and increased cellular iron levels are two important hallmarks of PD, this suggests a novel potential mechanism by which glutathione depletion may affect these processes in dopaminergic neurons. We report that depletion of glutathione as substrate results in a dose-dependent Grx2 inhibition and decreased iron incorporation into a mitochondrial complex I (CI) and aconitase (m-aconitase). Mitochondrial Grx2 inhibition through siRNA results in a corresponding decrease in CI and m-aconitase activities. It also results in significant increases in iron-regulatory protein (IRP) binding, likely as a consequence of conversion of Fe-S-containing cellular aconitase to its non-Fe-S-containing IRP1 form. This is accompanied by increased transferrin receptor, decreased ferritin, and subsequent increases in mitochondrial iron levels. This suggests that glutathione depletion may affect important pathologic cellular events associated with PD through its effects on Grx2 activity and mitochondrial Fe-S biogenesis. PMID- 19290778 TI - Nitroproteomics of peripheral blood mononuclear cells from patients and a rat model of ALS. AB - Increased levels of 3-nitrotyrosine in the central nervous system have been found in patients and mouse models of familial ALS (fALS), suggesting a possible use of nitrated proteins as biomarkers. We analyzed peripheral blood mononuclear cells (PBMCs), easily accessible samples, from sporadic ALS (sALS) patients and a rat model of fALS (a) to establish whether an increased level of nitrated proteins was present in PBMCs, too, and (b) to identify possible candidate biomarkers. With a proteomic approach, we identified for the first time the major overnitrated proteins in PBMCs from patients and rats at different disease stages. In the rats, their increased levels already were measured at a presymptomatic stage. Among them, actin, ATP synthase, and vinculin overlap between sALS patients and the rat model. Interestingly, in a previous study, actin and ATPase have been found overnitrated in the spinal cord of a mouse model of fALS before disease onset, suggesting their possible involvement in motor neuron degeneration. In conclusion, we observed that an increased level of nitrated proteins was not restricted to the spinal cord but also was present in peripheral cells of patients and an animal model, and that nitrated proteins are promising candidate biomarkers for early diagnosis of ALS. PMID- 19290779 TI - Determination of branched chain amino acids, methionine, phenylalanine, tyrosine and alpha-keto acids in plasma and dried blood samples using HPLC with fluorescence detection. AB - BACKGROUND: The determination of branched chain amino acids [BCAA; valine (Val), leucine (Leu), isoleucine (Ile)], alpha-keto acids derived from BCAA [BCKA; alpha ketoisovaleric acid (KIV), alpha-ketoisocaproic acid (KIC), alpha ketomethylvaleric acid (KMV)], methionine (Met), phenylalanine (Phe) and tyrosine (Tyr) is currently the most reliable approach for the diagnosis of maple syrup urine disease (MSUD), hypermethioninemia, phenylketonuria (PKU) and tyrosinemia. The aim of this study was to develop rapid and simple HPLC methods for measurement of BCAA, Met, Phe, Tyr and BCKA in plasma and dried blood samples. METHODS: Samples of peripheral venous blood with EDTA as anticoagulant were obtained from a group of healthy blood donors (n=70, 35 females, 27-41 years of age and 35 males, 28-43 years of age). Blood-spot samples from a group of newborns (n=80, 40 girls and 40 boys 3-5 days of age) were collected onto #903 Specimen Collection Paper and allowed to dry for at least 24 h before analysis. Prior to separation, the amino acids (AA) were derivatized with o phthaldialdehyde (OPA) and BCKA with o-phenylenediamine (OPD). Reverse phase column chromatography (LiChroCart 125-4 Purospher RP-18e, 5 microm) was used for separation and fluorescence detection used to monitoring of effluent. For AA analysis, 25 mmol/L sodium hydrogenphosphate-methanol (90:10, v/v), pH 6.5+/-0.1 was used as mobile phase A and 100% methanol was used as mobile phase B. Measurement of BCKA used a mixture of methanol and deionized water (55:45, v/v) as mobile phase A and mobile phase B consisted of 100% methanol. RESULTS: Analytical performance of these methods was satisfactory for the determination of all AA and BCKA. The intra-assay and inter-assay coefficients of variation were below 10% and recovery ranged from 90%-110%. CONCLUSIONS: We have developed simple, rapid and selective HPLC methods with fluorescence detection for the determination of BCAA, Met, Phe, Tyr and BCKA in plasma and dried blood samples. PMID- 19290780 TI - Efficacy, tolerability and safety of the fixed combination of bimatoprost 0.03% and timolol 0.5% in a broad patient population: multicenter, open-label observational study. AB - OBJECTIVE: To evaluate intraocular pressure (IOP)-lowering efficacy, tolerability, and safety of the fixed combination of bimatoprost 0.03% and timolol 0.5% (Ganfort) among German patients. METHODS: Multicenter, observational, open-label study of patients with primary open angle glaucoma or ocular hypertension (n = 606). As determined by participating physicians, patients had insufficient IOP control and required a medication change. They were switched to once-daily fixed-combination bimatoprost/timolol with no wash-out period. IOP was recorded at treated baseline, 4-6 weeks and 12 weeks after switching. Tolerability was measured using a 4-step scale (excellent, good, moderate, poor) and all adverse events were recorded. RESULTS: A total of 405 patients switched from monotherapy, 97 switched from other fixed combinations, and 104 switched from non-fixed combinations. Among all patients, 32.5% had used prostaglandin analog (PGA) monotherapy, 8.7% had been using a fixed combination that included a PGA, and 6.9% had been using an adjunctive combination of a PGA and a beta-blocker. Mean treated baseline IOP (+/-SD) for all patients was 20.7 +/- 3.5 mmHg. Overall, changing medication to fixed-combination bimatoprost/timolol lowered IOP to 16.6 +/- 2.7 mmHg (p < 0.001 vs. baseline) after 4-6 weeks and to 16.1 +/- 2.6 mmHg (p < 0.001) after 12 weeks; reductions of 19.8% and 22.2%, respectively. Combined bimatoprost/timolol provided an additional IOP reduction versus baseline in most subgroups based on prior treatment. At week 12, patients who had previously used a beta-blocker achieved an additional 25.8% decrease from baseline and IOP was reduced by 22.6% in former PGA monotherapy patients. At week 12, 84.6% of all eyes reached a target pressure less than or equal to 18 mmHg. Tolerability of bimatoprost/timolol was rated excellent or good by the physicians for 98.7% of patients and by 96.7% of the patients themselves. Few adverse events occurred during the treatment period. CONCLUSIONS: Although this study was limited by its observational design, our results show that the fixed combination of bimatoprost 0.03%/timolol 0.5% was effective, well tolerated, and safe in a broad patient population. PMID- 19290786 TI - Pharmacogenetics of the organic anion transporting polypeptide 1A2. AB - The solute carrier, human organic anion transporting polypeptide 1A2 (OATP1A2, OATP-A, OATP1 and OATP) is highly expressed in the intestine, kidney, cholangiocytes and the blood-brain barrier. This localization suggests that OATP1A2 may be vitally important in the absorption, distribution and excretion of a broad array of clinically important drugs. Several nonsynonymous polymorphisms have been identified in the gene encoding OATP1A2, SLCO1A2 (SLC21A3), with some of these variants demonstrating functional changes in the transport of OATP1A2 substrates. PMID- 19290787 TI - Novel DNA sequence variations of cytochrome P450 genes in the Han Chinese population. AB - AIM: Cytochrome P450 is an important monooxygenase responsible for the metabolism of a large variety of structurally diverse compounds. The aim of this study was to systematically investigate the DNA sequence variations in 14 cytochrome P450 genes relevant to drug metabolism in the Han Chinese population. MATERIALS & METHODS: We sequenced these 14 genes in 23 subjects and determined the allele frequencies. RESULTS: We identified a total of 312 genetic variants, which included 80 (25.6%) novel variants. These novel variants included 67 noncoding variants, four synonymous and nine nonsynonymous variants. Among these variants, an 11-nucleotide insertion in the 3-flanking site of intron 6 in CYP2E1 (c.968 77_-76insGATGGGTGGAT) had the highest allele frequency of 0.565. A total of 16 of these novel variants were predicted to have potential functional consequences; however, among them only c.-1299T>A in CYP2C18 and c.-498C>A in CYP2D6 reached a frequency of 4.9%. CONCLUSIONS: This study establishes a genetic database of cytochrome P450 genes in the Han Chinese population and suggests further genetic diversity throughout this important gene family. PMID- 19290788 TI - Association of CRTH2 gene polymorphisms with the required dose of antihistamines in patients with chronic urticaria. AB - INTRODUCTION: Chronic urticaria (CU), defined as the recurring incidence of wheals with or without angioedema for more than 6 weeks, is a common disorder associated with mast cell activation, degranulation, and histamine release. Considering the association between the CRTH2 gene and mast cells, we investigated the association of this gene polymorphism with the CU phenotype and antihistamine drug requirement in patients with CU. MATERIALS & METHODS: Two groups consisting of 384 patients with CU and 231 patients as normal controls (NCs) were enrolled from the Department of Allergy and Rheumatology, Ajou University Hospital, Suwon, Korea. Two polymorphisms of the CRTH2 gene, -466T>C and -129C>A were genotyped using primer extension methods. RESULTS: No significant differences were detected in the genotype and allele frequencies of the two CRTH2 polymorphisms between the CU and NC groups, and no significant associations were observed with clinical parameters, such as atopy status, serum total IgE, prevalence of autoantibodies and duration of CU. However, CU patients with homozygous TT genotypes had significantly higher dose requirements of antihistamines to control the CU symptoms (164.56 +/- 115.62 vs 137.38 +/- 90.15 loratadine equivalents, mg/week) than those with the CT and CC genotypes (p = 0.025). The luciferase activity was significantly enhanced in the construct containing CRTH2 466C compared with the -466T-containing construct (p < 0.001). Co-transfection experiments with GATA-3 (300 ng) and the -466T and -466C CRTH2 alleles revealed that the CRTH2 -466T allele produced a greater increase in induction of luciferase activity than the -466C allele (p < 0.001). CONCLUSION: The CRTH2 -466T>C gene polymorphism may not affect on the phenotype of CU, but contributes to the required dose of antihistamines in patients with CU. PMID- 19290789 TI - Association studies of catechol-O-methyltransferase (COMT) gene with schizophrenia and response to antipsychotic treatment. AB - AIM: We investigated the catechol-O-methyltrasferase (COMT) gene, which is a strong functional and positional candidate gene for schizophrenia and therapeutic response to antipsychotic medication. MATERIALS & METHODS: Single-locus as well as detailed haplotype-based association analysis of the COMT gene with schizophrenia and antipsychotic treatment response was carried out using seven COMT polymorphisms in 398 schizophrenia patients and 241 healthy individuals from a homogeneous south Indian population. Further responsiveness to risperidone treatment was assessed in 117 schizophrenia patients using Clinical Global Impressions (CGI). A total of 69 patients with a CGI score of 2 or less met the criteria of good responders and 48 were patients who continued to have a score of 3 and above and were classified as poor responders to risperidone treatment. RESULTS: The association of SNP rs4680 with schizophrenia did not remain significant after adjusting for multiple testing. Haplotype analysis showed highly significant association of seven COMT marker haplotypes with schizophrenia (CLUMP T4 p-value = 0.0001). Our results also demonstrated initial significant allelic associations of two SNPs with drug response (rs4633: chi(2) = 4.36, p value = 0.036, OR: 1.80, 95% CI: 1.03-3.15; and rs4680: chi(2) = 4.02, p-value = 0.044, OR: 1.76, 95% CI: 1.01-3.06) before multiple correction. We employed two marker sliding window analysis for haplotype association and observed a significant association of markers located between intron 1 and intron 2 (rs737865, rs6269: CLUMP T4 p-value = 0.021); and in exon 4 (rs4818, rs4680: CLUMP T4 p-value = 0.028) with drug response. CONCLUSION: The present study thus indicates that the interacting effects within the COMT gene polymorphisms may influence the disease status and response to risperidone in schizophrenia patients. However, the study needs to be replicated in a larger sample set for confirmation, followed by functional studies. PMID- 19290790 TI - MicroRNA polymorphisms: the future of pharmacogenomics, molecular epidemiology and individualized medicine. AB - Referred to as the micromanagers of gene expression, microRNAs (miRNAs) are evolutionarily conserved small noncoding RNAs. Polymorphisms in the miRNA pathway (miR-polymorphisms) are emerging as powerful tools to study the biology of a disease and have the potential to be used in disease prognosis and diagnosis. Detection of miR-polymorphisms holds promise in the field of miRNA pharmacogenomics, molecular epidemiology and for individualized medicine. MiRNA pharmacogenomics can be defined as the study of miRNAs and polymorphisms affecting miRNA function in order to predict drug behavior and to improve drug efficacy. Advancements in the miRNA field indicate the clear involvement of miRNAs and genetic variations within the miRNA pathway in the progression and prognosis of diseases such as cancer, neurological disorders, muscular hypertrophy, gastric mucosal atrophy, cardiovascular disease and Type II diabetes. Various algorithms are available to predict miRNA-target mRNA sites; however, it is advisable to use multiple algorithms to confirm the predictions. Polymorphisms that may potentially affect miRNA-mediated regulation of the cell can be present not only in the 3 -UTR of a miRNA target gene, but also in the genes involved in miRNA biogenesis and in pri-, pre- and mature-miRNA sequences. A polymorphism in processed miRNAs may affect expression of several genes and have serious consequences, whereas a polymorphism in miRNA target site, in the 3 UTR of the target mRNA, may be more target and/or pathway specific. In this review, we for the first time suggest a classification of miRNA polymorphisms/mutations. We also describe the importance and implications of miR polymorphisms in gene regulation, disease progression, pharmacogenomics and molecular epidemiology. PMID- 19290791 TI - Update on the pharmacogenetics of the vitamin D receptor and osteoporosis. AB - Vitamin D and calcium are essential for normal skeletal growth and for maintaining the mechanical and structural integrity of the skeleton. Reduced intake of calcium and vitamin D may be associated with reduced bone mass and osteoporosis while a chronic and severe vitamin D deficiency may lead to osteomalacia. Given the importance of vitamin D in bone homeostasis, common polymorphisms in the vitamin D receptor gene were the first to be investigated as possible determinants of bone mass and fracture risk. Even though results are still conflicting and the molecular mechanisms by which these polymorphisms influence receptor activity remain in part to be investigated, an additional important issue is represented by their potential pharmacogenomic and pharmacogenetic implications. This review analyzes major pharmacogenetic studies of polymorphisms in the vitamin D receptor gene and osteoporosis. PMID- 19290793 TI - Cutaneous leishmaniasis: disease susceptibility and pharmacogenetic implications. AB - Cutaneous leishmaniasis is a major tropical infection of public health importance caused by a number of vector-borne Leishmania protozoa species. Evidence supports a highly complex etiology. Environmental, parasite and host factors determine pathogenesis, and result in a diverse clinical spectrum of disease. Disease susceptibility, clinical course, prognosis and therapy response are highly variable, suggesting a genetic basis. Epidemiological studies have demonstrated familial aggregation, and family and association studies have identified HLA and non-HLA gene associations. Further progress in susceptibility gene identification for leishmaniasis would require genome-wide scans and candidate gene-association studies in large cohorts. Correlation between host genotype and therapy response has important pharmacogenetic implications, especially as current therapies for leishmaniasis are inadequate and progress in new drug development has been poor. PMID- 19290792 TI - Application of pharmacogenomics to malaria: a holistic approach for successful chemotherapy. AB - Drug resistance in malaria jeopardizes the most elementary objectives of malaria control--reducing suffering and eliminating mortality. An important, and so far the only known, mechanism of drug resistance appears to be polymorphisms in the malaria parasite genes. Efforts to circumvent antimalarial drug resistance now range from the use of combination therapies with existing agents to genomics based studies directed toward discovering novel targets and agents. However, the potential contribution of host genetic/molecular factors, particularly those associated with antimalarial drug metabolism, remains largely unexplored. Our knowledge concerning the basic mechanisms involved in the pharmacokinetics of antimalarial drugs is fragmentary. In addition, the link between antimalarial drug pharmacokinetics and treatment outcomes is generally unclear. The purpose of this article is to provide general background information on antimalarial drug resistance and associated parasite genetic factors, and subsequently highlight the aforementioned unexplored and unclear areas, with a view to stimulate much needed further research. PMID- 19290794 TI - Significance of genetic polymorphisms of the renin-angiotensin-aldosterone system in cardiovascular and renal disease. AB - The angiotensin converting enzyme (ACE) is a component of the renin-angiotensin aldosterone system (RAAS). The RAAS--involved primarily in blood pressure and sodium homeostasis--is activated in many renal and cardiovascular diseases, and therapy with ACE inhibitors and other blockers of the RAAS has proven to be clinically beneficial. Plasma and tissue levels of ACE are at least partially determined by a genetic polymorphism based on the presence (insertion [I]) or absence (deletion [D]) of a 287 base pair element in intron 16. In particular Asian subjects with the DD genotype (and increased ACE activity) have been reported to be at higher risk for cardiovascular disorders and nephropathy. Numerous studies evaluated the role of the ACE I/D polymorphism as well as other genetic variants of the RAAS in the context of RAAS inhibitor therapy. However, as race and environmental factors, such as salt intake also affect treatment response most studies were underpowered leading to conflicting results. PMID- 19290795 TI - CYP3A5 and ABCB1 genes and hypertension. AB - Hypertension is the first single modifiable cause of disease burden worldwide. Genes encoding proteins that are involved in the metabolism (CYP3A5) and transport (ABCB1) of drugs and hormones might contribute to blood pressure control in humans. Indeed, recent data have suggested that CYP3A5 and ABCB1 gene polymorphisms are associated with blood pressure in the rat as well as in humans. Interestingly, the effects of these genes on blood pressure appear to be modified by dietary salt intake. This review summarizes what is known regarding the relationships of the ABCB1 and CYP3A5 genes with blood pressure, and discusses the potential underlying mechanisms of the association. If the role of these genes in blood pressure control is confirmed in other populations and other ethnic groups, these findings would point toward a new pathway for blood pressure control in humans. PMID- 19290796 TI - Visual genotyping of SNPs of drug-metabolizing enzymes by tetra-primer PCR coupled with a dry-reagent DNA biosensor. AB - BACKGROUND: SNP-typing strategies involve an exponential amplification step, an allele discrimination reaction and detection of the products. Usually, allele discrimination is performed after amplification. Tetra-primer PCR allows allele discrimination during the amplification step, thereby avoiding additional genotyping reactions. However, to date, electrophoresis is the only method used for detection of tetra-primer PCR products. We report a dipstick test that enables visual detection of tetra-primer PCR products within minutes without instruments. The method is applied to the genotyping of CYP2C19*2 (c.681G>A) and CYP2D6*4 (g.3465G>A). MATERIALS & METHODS: A pair of external primers amplifies a segment encompassing the SNPs. Biotinylated inner primers have a 3 -mismatch and pair off with the external primers to guide a bidirectional amplification that generates allele-specific fragments. The products are hybridized briefly with poly(dA)-tailed probes and applied to the DNA biosensor, which is then immersed in the appropriate buffer. As the buffer migrates along the biosensor, the hybrids are captured from streptavidin at the test zone and interact with oligo(dT)-functionalized gold nanoparticles leading to the formation of a red line. Another red line is formed at the control zone to indicate proper function of the sensor. RESULTS: We genotyped 55 samples for CYP2C19*2 and 49 samples for CYP2D6*4. The accuracy of this method was confirmed by sequencing and electrophoresis. CONCLUSIONS: The unique advantages of the proposed method are its simplicity and low cost. Contrary to electrophoresis, hybridization provides sequence confirmation of amplified fragments. The dry-reagent dipstick format minimizes the requirements for highly qualified personnel. PMID- 19290799 TI - Micro-CT of corrosion casts for use in the computer-aided design of microvasculature. AB - Two-dimensional micro-computed tomography (micro-CT) slices can be reconstructed into three-dimensional (3D) models that demonstrate capillary beds. This study focused on the acquisition of data necessary to create scaffolding that directly mimics the unique structural patterns of a microvascular tree system. The Microfil vascular contrasting method was compared to the Baston's methylmethacrylate corrosion casting (BMCC) method to determine which provided the most accurate and high-resolution results for 3D micro-CT reconstruction derived from the two-dimensional micro-CT slices of the capillary beds. It was determined that the BMCC, a method traditionally used in the scanning electron microscopic analysis of the microvasculature, was the best method for representing capillary lumina for micro-CT scanning. The removal of tissues from the BMCC cast resulted in samples that eliminated background material, thus increasing the X-ray contrast levels of the CT images. This provided for a more complete and more distinguishable high-resolution image of the represented capillary lumina. Images created with this BMCC method were reconstructed in a stereolithography file format as 3D mesh structure for later importing into computer-aided design (CAD) software. The resulting Bio-CAD, then, can be used to guide the more accurate fabrication of the microvascular scaffolding and then serve as the framework for tissue engineering of microvascular structures. Results from this study clearly indicated that the BMCC method is superior to the Microfil method for accurate and complete high-resolution imaging of capillary beds. PMID- 19290800 TI - Noninvasive monitoring of embryonic stem cells in vivo with MRI transgene reporter. AB - Reporter gene-based magnetic resonance imaging (MRI) offers unique insights into behavior of cells after transplantation, which could significantly benefit stem cell research and translation. Several candidate MRI reporter genes, including one that encodes for iron storage protein ferritin, have been reported, and their potential applications in embryonic stem (ES) cell research have yet to be explored. We have established transgenic mouse ES (mES) cell lines carrying human ferritin heavy chain (FTH) as a reporter gene and succeeded in monitoring the cell grafts in vivo using T(2)-weighted MRI sequences. FTH generated MRI contrast through compensatory upregulation of transferrin receptor (Tfrc) that led to increased cellular iron stored in ferritin-bound form. At a level sufficient for MRI contrast, expression of FTH posed no toxicity to mES cells and did not interfere with stem cell pluripotency as observed in neural differentiation and teratoma formation. The compatibility and functionality of ferritin as a reporter in mES cells opens up the possibility of using MRI for longitudinal noninvasive monitoring of ES cell-derived cell grafts at both molecular and cellular levels. PMID- 19290801 TI - High-throughput and combinatorial technologies for tissue engineering applications. AB - As the field of tissue engineering progresses, new technology is essential to accelerate the identification of potentially translatable approaches for the repair of tissues damaged due to disease or trauma. The development of high throughput and combinatorial technologies is helping to speed up research that is applicable to all aspects of the tissue engineering paradigm. This diverse technology can be used for both the rapid synthesis of polymers and their characterization with respect to local and bulk properties in a high-throughput fashion. The interactions of cells with many diverse materials in both two- and three-dimensions can be assessed rapidly through the use of microarrays and rapid outcome measures and with microfluidic devices for investigation of soluble factor and material combinations. Finally, small molecule screening of large libraries is being used to identify molecules that exhibit control over cell behavior, including stem cell differentiation. All of these approaches are aimed to move beyond traditional iterative methods to identify unique materials and molecules that would not have been identified otherwise. Although much of this work is only applicable for in vitro studies, future methods may translate into rapid screening of these approaches in vivo. PMID- 19290802 TI - Human Vitronectin-Derived Peptide Covalently Grafted onto Titanium Surface Improves Osteogenic Activity: A Pilot In Vivo Study on Rabbits. AB - Peptide and protein exploitation for the biochemical functionalization of biomaterial surfaces allowed fabricating biomimetic devices able to evoke and promote specific and advantageous cell functions in vitro and in vivo. In particular, cell adhesion improvement to support the osseointegration of implantable devices has been thoroughly investigated. This study was aimed at checking the biological activity of the (351-359) human vitronectin precursor (HVP) sequence, mapped on the human vitronectin protein; the peptide was covalently linked to the surface of titanium cylinders, surgically inserted in the femurs of New Zealand white rabbits and analyzed at short experimental time points (4, 9, and 16 days after surgery). To assess the osteogenic activity of the peptide, three vital fluorochromic bone markers were used (calcein green, xylenol orange, and calcein blue) to stain the areas of newly grown bone. Static and dynamic histomorphometric parameters were measured at the bone-implant interface and at different distances from the surface. The biological role of the (351-359)HVP sequence was checked by comparing peptide-grafted samples and controls, analyzing how and how much its effects change with time across the bone regions surrounding the implant surface. The results obtained reveal a major activity of the investigated peptide 4 days after surgery, within the bone region closest to the implant surface, and larger bone to implant contact 9 and 16 days after surgery. Thus, improved primary fixation of endosseous devices can be foreseen, resulting in an increased osteointegration. PMID- 19290803 TI - Linear ordered collagen scaffolds loaded with collagen-binding brain-derived neurotrophic factor improve the recovery of spinal cord injury in rats. AB - Growth factors such as brain-derived neurotrophic factor (BDNF) are widely used in the recovery of spinal cord injury (SCI) for promoting axonal regeneration or improving neuron survival. However, the lack of efficient delivery approaches limits their clinical applications. In this study, we constructed an efficient delivery system including collagen-binding BDNF and linear ordered collagen scaffolds (LOCS). A collagen-binding domain (CBD) was utilized to construct collagen-targeting BDNF (CBD-BDNF) to allow its specific binding to the collagen. In vitro activity assay showed that CBD-BDNF had similar bioactivity in neurite outgrowth of dorsal root ganglia and the survival of PC12 cells. CBD-BDNF was shown to have specific binding activity to collagen. Using the rat hemisection SCI model, we found that LOCS loaded with CBD-BDNF significantly improved the SCI recovery evaluated by the Basso, Beattie, and Bresnahan scale and immunohistochemical staining with anti-neurofilament antibody. Thus, this targeting drug delivery system consisting of CBD-BDNF and LOCS could be an effective strategy for the repair of SCI. PMID- 19290804 TI - The effect of hydrostatic pressure on three-dimensional chondroinduction of human adipose-derived stem cells. AB - BACKGROUND: The optimal production of three-dimensional cartilage in vitro requires both inductive factors and specified culture conditions (e.g., hydrostatic pressure [HP], gas concentration, and nutrient supply) to promote cell viability and maintain phenotype. In this study, we optimized the conditions for human cartilage induction using human adipose-derived stem cells (ASCs), collagen scaffolds, and cyclic HP treatment. METHODS: Human ASCs underwent primary culture and three passages before being seeded into collagen scaffolds. These constructs were incubated for 1 week in an automated bioreactor using cyclic HP at 0-0.5 MPa, 0.5 Hz, and compared to constructs exposed to atmospheric pressure. In both groups, chondrogenic differentiation medium including transforming growth factor-beta1 was employed. One, 2, 3, and 4 weeks after incubation, the cell constructs were harvested for histological, immunohistochemical, and gene expression evaluation. RESULTS: In histological and immunohistochemical analyzes, pericellular and extracellular metachromatic matrix was observed in both groups and increased over 4 weeks, but accumulated at a higher rate in the HP group. Cell number was maintained in the HP group over 4 weeks but decreased after 2 weeks in the atmospheric pressure group. Chondrogenic specific gene expression of type II and X collagen, aggrecan, and SRY-box9 was increased in the HP group especially after 2 weeks. CONCLUSION: Our results demonstrate chondrogenic differentiation of ASCs in a three-dimensional collagen scaffolds with treatment of a cyclic HP. Cyclic HP was effective in enhancing accumulation of extracellular matrix and expression of genes indicative of chondrogenic differentiation. PMID- 19290805 TI - Mesenchymal stem cells derived from canine umbilical cord vein--a novel source for cell therapy studies. AB - The canine model provides a large animal system to evaluate many treatment modalities using stem cells (SCs). However, only bone marrow (BM) protocols have been widely used in dogs for preclinical approaches. BM donation consists of an invasive procedure and the number and differentiation potential of its mesenchymal stem cells (MSCs) decline with age. More recently, umbilical cord was introduced as an alternative source to BM since it is obtained from a sample that is routinely discarded. Here, we describe the isolation of MSCs from canine umbilical cord vein (cUCV). These cells can be obtained from every cord received and grow successfully in culture. Their multipotent plasticity was demonstrated by their capacity to differentiate in adipocytic, chondrocytic, and osteocytic lineages. Furthermore, our results open possibilities to use cUCV cells in preclinical trials for many well-characterized canine model conditions homologs to human diseases. PMID- 19290806 TI - Smooth muscle cell seeding of decellularized scaffolds: the importance of bioreactor preconditioning to development of a more native architecture for tissue-engineered blood vessels. AB - Vascular smooth muscle cells (VSMCs) impart important functional characteristics in the native artery and, therefore, should logically be incorporated in the development of tissue-engineered blood vessels. However, the native architecture and low porosity of naturally derived biomaterials (i.e., decellularized vessels) have impeded efforts to seed and incorporate a VSMC layer in tissue-engineered blood vessels. To this end, the goal of this study was to develop improved methods for seeding, proliferation, and maturation of VSMCs on decellularized porcine carotid arteries. Decellularized vessels were prepared in the absence and presence of the adventitial layer, and statically seeded with a pipette containing a suspension of rat aortic VSMCs. After cell seeding, recellularized engineered vessels were placed in a custom bioreactor system for 1-2 weeks to enhance cellular proliferation, alignment, and maturation. Initial attachment of VSMCs was dramatically enhanced by removing the adventitial layer of the decellularized porcine artery. Moreover, cyclic bioreactor conditioning (i.e., flow and pressure) resulted in increased VSMC proliferation and accelerated formation of a muscularized medial layer in the absence of the adventitial layer during the first week of preconditioning. Fura-2-based digital imaging microscopy revealed marked and reproducible depolarization-induced calcium mobilization after bioreactor preconditioning in the absence, but not in the presence, of the adventitia. The major finding of this investigation is that bioreactor preconditioning accelerates the formation of a significant muscular layer on decellularized scaffolds, in particular on adventitia-denuded scaffolds. Further, the VSMC layer of bioreactor-preconditioned vessels was capable of mobilizing calcium in response to cellular depolarization. These findings represent an important first step toward the development of tissue-engineered vascular grafts that more closely mimic native vasculature. PMID- 19290807 TI - Genome-environment interactions and prospective technology assessment: evolution from pharmacogenomics to nutrigenomics and ecogenomics. AB - The relationships between food, nutrition science, and health outcomes have been mapped over the past century. Genomic variation among individuals and populations is a new factor that enriches and challenges our understanding of these complex relationships. Hence, the confluence of nutritional science and genomics nutrigenomics--was the focus of the OMICS: A Journal of Integrative Biology in December 2008 (Part 1). The 2009 Special Issue (Part 2) concludes the analysis of nutrigenomics research and innovations. Together, these two issues expand the scope and depth of critical scholarship in nutrigenomics, in keeping with an integrated multidisciplinary analysis across the bioscience, omics technology, social, ethical, intellectual property and policy dimensions. Historically, the field of pharmacogenetics provided the first examples of specifically identifiable gene variants predisposing to unexpected responses to drugs since the 1950s. Brewer coined the term ecogenetics in 1971 to broaden the concept of gene-environment interactions from drugs and nutrition to include environmental agents in general. In the mid-1990s, introduction of high-throughput technologies led to the terms pharmacogenomics, nutrigenomics and ecogenomics to describe, respectively, the contribution of genomic variability to differential responses to drugs, food, and environment defined in the broadest sense. The distinctions, if any, between these newer fields (e.g., nutrigenomics) and their predecessors (e.g., nutrigenetics) remain to be delineated. For nutrigenomics, its reliance on genome-wide analyses may lead to detection of new biological mechanisms governing host response to food. Recognizing "genome-environment interactions" as the conceptual thread that connects and runs through pharmacogenomics, nutrigenomics, and ecogenomics may contribute toward anticipatory governance and prospective real-time analysis of these omics fields. Such real-time analysis of omics technologies and innovations is crucial, because it can influence and positively shape them as these approaches develop, and help avoid predictable pitfalls, and thus ensure their effective and ethical application in the laboratory, clinic, and society. PMID- 19290808 TI - Mononuclear cell transcriptome response after sustained virgin olive oil consumption in humans: an exploratory nutrigenomics study. AB - Virgin olive oil (VOO) is considered to be one of the main components responsible for the health benefits of the Mediterranean diet, particularly against atherosclerosis where peripheral blood mononuclear cells (PBMNCs) play a crucial role in atherosclerosis development and progression. The objective of this article was to identify the PBMNC genes that respond to VOO consumption in order to ascertain the molecular mechanisms underlying the beneficial action of VOO in the prevention of atherosclerosis. Gene expression profiles of PBMNCs from healthy individuals were examined in pooled RNA samples by microarrays after 3 weeks of moderate and regular consumption of VOO, as the main fat source in a diet controlled for antioxidant content. Gene expression was verified by qPCR. The response to VOO consumption was confirmed for individual samples (n = 10) by qPCR for 10 upregulated genes (ADAM17, ALDH1A1, BIRC1, ERCC5, LIAS, OGT, PPARBP, TNFSF10, USP48, and XRCC5). Their putative role in the molecular mechanisms involved in atherosclerosis development and progression is discussed, focusing on a possible relation with VOO consumption. Our data support the hypothesis that 3 weeks of nutritional intervention with VOO supplementation, at doses common in the Mediterranean diet, can alter the expression of genes related to atherosclerosis development and progression. PMID- 19290809 TI - The STEDMAN project: biophysical, biochemical and metabolic effects of a behavioral weight loss intervention during weight loss, maintenance, and regain. AB - The Study of the Effects of Diet on Metabolism and Nutrition (STEDMAN) Project uses comprehensive metabolic profiling to probe biochemical mechanisms of weight loss in humans. Measurements at baseline, 2 and 4 weeks, 6 and 12 months included diet, body composition, metabolic rate, hormones, and 80 intermediary metabolites measured by mass spectrometry. In 27 obese adults in a behavioral weight loss intervention, median weight decreased 13.9 lb over the first 6 months, then reverted towards baseline by 12 months. Insulin resistance (HOMA) was partially ameliorated in the first 6 months and showed sustained improvement at 12 months despite weight regain. Ghrelin increased with weight loss and reverted to baseline, whereas leptin and PYY fell at 6 months and remained persistently low. NPY levels did not change. Factors possibly contributing to sustained improvement in insulin sensitivity despite weight regain include adiponectin (increased by 12 months), IGF-1 (increased during weight loss and continued to increase during weight regain), and visceral fat (fell at 6 months but did not change thereafter). We observed a persistent reduction in free fatty acids, branched chain amino acids, and related metabolites that may contribute to improved insulin action. These findings provide evidence for sustained benefits of weight loss in obese humans and insights into mechanisms. PMID- 19290810 TI - Knowledge and attitudes of Canadian consumers and health care professionals regarding nutritional genomics. AB - With advances in the field of nutrigenomics, commercial laboratories have begun marketing genotyping services, nutritional advice, and dietary supplements "tailored" to match individual genetic predispositions. Although primarily offered by American companies, these services are available to Canadian consumers via the Internet. Qualitative research in the form of focus groups with members of the Canadian public was undertaken to assess the current level of understanding of and receptivity toward this new genomic application. Additionally, focus groups with health care professionals (physicians, pharmacists, dieticians, nutritionists, and naturopaths) investigated their interest in integrating nutrigenomics into health care delivery, and their capacity to do so. Gauging knowledge and attitudes early in the introduction of a new technology serves to identify potential "blind spots" regarding the ethical, legal, and social implications. Preliminary results indicate consumers believe potential benefits of nutrigenomics outweigh risks, while health care professionals express more skepticism. Both groups agree that more public education about nutrigenomics is needed and that regulatory oversight should ensure consumer protection. PMID- 19290812 TI - Nutrigenomics patents and commercialization: old wine in a new bottle? AB - This paper looks at the ethical, legal, and social issues (ELSI) associated with commercialization and patenting through the lens of nutrigenomics. These are two areas have generated a great deal of ELSI literature, although very little specific to nutrigenomic research. Nutrigenomic researchers seem likely to face the same patent concerns as those associated with gene patents more generally- specifically, that patents will hurt research and the distribution and uptake of useful technologies. Likewise, there is concern that commercialization pressure will lead to the inappropriate and premature implementation of nutrigenomic services. This paper concludes that while the patenting issues do not seem unique or particularly worrisome in the context of nutrigenomics, the early commercialization of testing is cause for concern and worthy of careful policy consideration. PMID- 19290811 TI - Risk assessment and communication tools for genotype associations with multifactorial phenotypes: the concept of "edge effect" and cultivating an ethical bridge between omics innovations and society. AB - Applications of omics technologies in the postgenomics era swiftly expanded from rare monogenic disorders to multifactorial common complex diseases, pharmacogenomics, and personalized medicine. Already, there are signposts indicative of further omics technology investment in nutritional sciences (nutrigenomics), environmental health/ecology (ecogenomics), and agriculture (agrigenomics). Genotype-phenotype association studies are a centerpiece of translational research in omics science. Yet scientific and ethical standards and ways to assess and communicate risk information obtained from association studies have been neglected to date. This is a significant gap because association studies decisively influence which genetic loci become genetic tests in the clinic or products in the genetic test marketplace. A growing challenge concerns the interpretation of large overlap typically observed in distribution of quantitative traits in a genetic association study with a polygenic/multifactorial phenotype. To remedy the shortage of risk assessment and communication tools for association studies, this paper presents the concept of edge effect. That is, the shift in population edges of a multifactorial quantitative phenotype is a more sensitive measure (than population averages) to gauge the population level impact and by extension, policy significance of an omics marker. Empirical application of the edge effect concept is illustrated using an original analysis of warfarin pharmacogenomics and the VKORC1 genetic variation in a Brazilian population sample. These edge effect analyses are examined in relation to regulatory guidance development for association studies. We explain that omics science transcends the conventional laboratory bench space and includes a highly heterogeneous cast of stakeholders in society who have a plurality of interests that are often in conflict. Hence, communication of risk information in diagnostic medicine also demands attention to processes involved in production of knowledge and human values embedded in scientific practice, for example, why, how, by whom, and to what ends association studies are conducted, and standards are developed (or not). To ensure sustainability of omics innovations and forecast their trajectory, we need interventions to bridge the gap between omics laboratory and society. Appreciation of scholarship in history of omics science is one remedy to responsibly learn from the past to ensure a sustainable future in omics fields, both emerging (nutrigenomics, ecogenomics), and those that are more established (pharmacogenomics). Another measure to build public trust and sustainability of omics fields could be legislative initiatives to create a multidisciplinary oversight body, at arm's length from conflict of interests, to carry out independent, impartial, and transparent innovation analyses and prospective technology assessment. PMID- 19290841 TI - Chitosan nanoparticles for drug delivery to the eye. AB - The purpose of this review is to provide the reader with an overview of the advances made in ocular delivery of bioactive molecules by means of chitosan based nanosystems, and their potential relevance in clinical use. The studies described clearly emphasise that chitosan-based nanostructures are versatile systems that can be tailor-made according to required compositions, surface characteristics and particle size. Such parameters, which are known to influence their in vivo performance, can be modulated by adjusting the formulation conditions of the nanotechnologies responsible for their formation, by incorporating additional materials in the preparation steps, and/or by using synthetically modified chitosan. Moreover, this review illustrates how the advances achieved in the understanding of the interaction of nanosystems with the ocular structures should result in the coming years, logically, into challenging innovations in ocular nanomedicines with significant impact on clinical practice. PMID- 19290842 TI - Challenges and opportunities in CNS delivery of therapeutics for neurodegenerative diseases. AB - With an increase in lifespan and changing population demographics, the incidence of central nervous system (CNS) diseases is expected to increase significantly in the 21st century. The most challenging of the CNS diseases are neurodegenerative diseases, characterized by age-related gradual decline in neurological function, often accompanied by neuronal death. Alzheimer's disease, Parkinson's disease and Huntington's disease are some examples of neurodegenerative diseases and have been well described in terms of disease mechanisms and pathology. However, successful treatment strategies for neurodegenerative diseases have so far been limited. Delivery of drugs into the CNS is one of the most challenging problems faced in the treatment of neurodegeneration. In this review, we describe the difficulties with CNS therapy, especially with the use of biological macromolecules, such as proteins and nucleic acid constructs. CNS therapeutics also represents a huge opportunity and examples of strategies that can enhance therapeutic delivery for the treatment of neurodegenerative diseases are emphasized. It is anticipated that with an increase in biological understanding of neurodegenerative diseases, there will be even more therapeutic opportunities. As such, these delivery strategies have a very important role to play in the future in the translation of CNS therapeutics from bench to bedside. PMID- 19290843 TI - Stability of selected serum proteins after long-term storage in the Janus Serum Bank. AB - BACKGROUND: Human serum from biobanks is frequently used in prospective epidemiological studies. Long-term storage may modify its composition. A better understanding of the stability of the serum components may improve the interpretation of future studies. METHODS: The concentrations of selected proteins; immunoglobulins, carrier proteins and enzymes in samples stored at -25 degrees C for 25 years and 2 years were compared with 1-month-old samples. For each length of storage time, 130 specimens were randomly selected from apparently healthy male blood donors aged 40-49 years. We examined the distribution of values, compared dispersion and localization of central tendency, and established reference intervals for each component. RESULTS: The study demonstrated non significant or numerically small group differences in the concentrations of albumin, aspartate amino transferase, cystatin C, immunoglobulin E, immunoglobulin G, and sex hormone binding globulin. Mean values between fresh and 25-year-old samples suggested larger differences during storage for alanine amino transferase (-73.4%), creatinine kinase (-96.1%), insulin C-peptide (-98.7%), ferritin (-18.5%) and transferrin (+8.2%). CONCLUSIONS: The findings showed that long-term storage can introduce a considerable bias for vulnerable components. PMID- 19290844 TI - Effect of maternal smoking on cord blood estriol, placental lactogen, chorionic gonadotropin, FSH, LH, and cortisol. AB - AIM: To determine the effect of maternal cigarette smoking on cord blood concentrations of E3, hPL, beta-hCG, FSH, LH, and cortisol. METHODS: Hormone concentrations were measured in term neonates of 100 smoking and 100 non-smoking mothers. RESULTS: The median E3, hPL, beta-hCG, FSH, LH and cortisol cord blood concentrations in the non-smoking mothers' offspring were 212 ng/mL, 2.00 microg/mL, 57.5 mIU/mL, 0.10 mIU/mL, 0.20 mIU/mL, and 14.3 microg/mL, respectively; in the smoking they were 163, 1.39, 45.4, 0.10, 0.20, and 25.1, respectively (P=0.008, 0.004, 0.037, 0.498, 0.286, 0.004, respectively). There was a significant but poor negative correlation between number of cigarettes/day and E3 (r=-0.163, P=0.021), hPL (r=-0.191, P=0.007), and beta-hCG (r=-0.143, P=0.044), whereas the correlation with cortisol was positive (r=0.259, P<0.0001). Multiple linear regression analyses showed that maternal smoking is a determinant of cord blood E3, hPL, beta-hCG, FSH, and cortisol concentrations. CONCLUSIONS: Tobacco smoking is associated with a reduction in cord blood E3, hPL, and beta hCG concentrations, whereas it is associated with increased cortisol concentrations. The disturbed endocrine equilibrium of the fetus induced by tobacco smoking could have adverse consequences on the fetus and child since fetal brain is a target organ for hormonal actions. PMID- 19290845 TI - Embryo vascularization by three-dimensional power Doppler ultrasonography at 7-10 weeks of pregnancy. AB - AIM: Assess vascular indices of 7-10 week embryos using three-dimensional power Doppler (3DPD) and correlate them with the crown-rump length (CRL). METHODS: This cross-sectional study included 65 healthy pregnancies between 7 and 10 weeks. The three-dimensional volume of the embryo was obtained using an endocavitary volumetric transducer and the VOCAL (Virtual Organ Computer-aided Analysis) method, with a 12 degree rotation angle and 15 sequential planes. The vascularization (VI), flow (FI) and the vascular and flow (VFI) indices were obtained using 3DPD and the mean, median, standard deviation, maximum and minimum values were calculated for each gestational age. Pearson's correlation coefficient (r) was used to evaluate the correlation between vascular indices and CRL. RESULTS: The VI ranged from 0.77 to 41.67, mean 14.68 (+/-8.60), the FI went from 25.71 to 139.50, mean 90.61 (+/-21.51) and the VFI from 0.20 to 81.57, mean 15.69 (+/-12.42). The correlation between CRL and all 3D power Doppler vascular indices was low (VI - r=-0.073, P=0.566; FI - r=0.173, P=0.168 and VFI - r= 0.004, P=0.974). CONCLUSION: 3D power Doppler vascular indices in 7-10 week embryos do not correlate with CRL. PMID- 19290846 TI - Obstetrical chondro-epiphyseal separation of the distal humerus: a case report and review of literature. PMID- 19290847 TI - Changes in symphysis pubis width during labor. AB - We studied changes in the width of the symphysis pubis in 32 women examined serially by ultrasound during labor. Measurements were made at the superior border of the symphysis and at its narrowest breadth in the latent phase, the active phase, and the second stage of labor. There was a significant increase in the width of the symphysis between the first and second stages of labor at both measured levels. Widening was observed in 94% at the superior symphyseal breadth and in 59% at the narrowest. Of those cases in which the width of the symphysis increased, there was a large spectrum of change, ranging from 9 to 98% of the original width at the narrowest measurement site and from 2 to 139% at the superior breadth. There was a strong inverse correlation between maternal age and the degree of symphyseal widening in nulliparas, but not in multiparas. We conclude that labor is associated with a substantial widening of the symphysis pubis in most, but not all women. PMID- 19290848 TI - Post-term pregnancy as a risk factor for intrapartum fetal distress. PMID- 19290849 TI - Detailed multigrade evaluation of fetal disorders with the quantified actocardiogram. AB - AIMS: To evaluate fetal disorders using detailed quantitative values from the actocardiogram (ACG) involving simultaneous tracing of ultrasonic Doppler fetal movement bursts and fetal heart rate (FHR). METHODS: Duration of FHR accelerations and fetal movement bursts were measured manually in 20 common fetal disorders. The severity of the fetal disorder was estimated using the FHR acceleration duration to movement burst ratio (A/B ratio) and 10-0 clinical severity ranks derived from the A/B ratio. The correlation of the A/B ratio and 1 and 5 min Apgar scores, as well as numerically expressed long-term outcomes were studied. RESULTS: A/B ratios were significantly correlated with the 1 and 5 min Apgar scores and the numerically evaluated long-term outcomes. Controversial cases of FHR pattern were more easily understood using the A/B ratio. The 10-0 severity derived from the A/B ratio was useful in clinical fetal studies. CONCLUSION: Common fetal disorders were evaluated quantitatively and in more detail using the A/B ratio from the actocardiogram than when using common binary good or bad evaluation. The A/B ratio was useful in outcome estimation, where the prognostic capability of the A/B ratio was confirmed by significant correlation with 1 and 5 min Apgar scores and long-term outcomes of fetal disorders. PMID- 19290850 TI - Women need more propofol than men during EEG-monitored total intravenous anaesthesia / Frauen benotigen mehr Propofol als Manner wahrend EEG-uberwachter total-intravenoser Anasthesie. AB - Gender-related differences in the pharmacology of drugs used in anaesthesiology have been reported by different authors. The aim of this study was to compare propofol dosages in a greater number of male and female patients who had received electroencephalogram (EEG) monitoring to maintain a defined depth of anaesthesia. Data from an EEG-controlled study were analysed with regard to gender differences in the consumption of the short-acting hypnotic propofol during maintenance of total intravenous anaesthesia and with regard to recovery times. The 656 patients (239 male, 417 female) were 15 to 97 years old, underwent different surgical procedures, and received propofol in combination with remifentanil, a short acting opioid. During the steady-state of anaesthesia the EEG stage D(2)/E(0), which corresponds to deep hypnosis, was the target level (EEG monitor: Narcotrend). Propofol dosages were calculated as mg/kg body weight/h and as mg/kg lean body mass/h. Significantly higher propofol dosages were observed in female patients compared to male patients, especially with lean body mass as a reference parameter. The dosages were characterised by a high interindividual variability. The time from stop of propofol until extubation was significantly shorter in women than in men. The propofol dosage for maintenance of anaesthesia at the EEG level D(2)/E(0) decreased with increasing age. PMID- 19290851 TI - Comparison and evaluation of existing methods for the extraction of low amplitude electrocardiographic signals: a possible approach to transabdominal fetal ECG. AB - Analysis of the fetal ECG (fECG) allows physicians to detect changes in the well being state of the fetus. But when assessing the fECG through the abdominal signals (ADS), its very low amplitude causes a problem, as the fECG representation in the ADS is buried in a mixture of other signals with stronger energy. Different methods have been proposed in the past to extract the transabdominal fECG for instantaneous fetal heart rate (fHR) computation; four representatives of them are selected for an accurate comparison of their performance in fECG extraction and in fHR estimation. A model for the ADS including all the possible disturbances is developed within this study to generate simulated data as they are required for the quantitative comparison of the algorithms. Their performances and limits considering both the enhancement of the fECG and the ability to preserve fECG morphology are analyzed using the simulated data. The results clearly show that linear methods for maternal ECG removal provide better results with respect to the extraction of the fECG morphology. The algorithms are then tested on real ADS data recorded during labor. Finally, the advantage of considering linear methods for ADS processing is discussed. PMID- 19290852 TI - Coagulation and fibrinolysis related cytokine imbalance in preeclampsia: the role of placental trophoblasts. AB - OBJECTIVE: Cytokine imbalance might have a pivotal role in hypercoagulability seen in preeclampsia. Our objective was to determine the relationship of blood coagulation related factors in placental tissue and peripheral blood in preeclamptic and normal pregnancies. METHODS: We compared mRNA and protein levels of tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), and tissue factor (TF) in the placenta of normal and preeclamptic pregnancies. Placental and peripheral blood t-PA and PAI-1 levels were examined. Trophoblasts were used to study the effects of hypoxia, hypoxia-reperfusion, and inflammatory cytokines on t-PA, PAI-1, tissue factor pathway inhibitor (TFPI), and TF. RESULTS: PAI-1 and TF mRNA and protein levels were higher in placental tissue of preeclamptic pregnancies and in the peripheral blood of patients with preeclampsia. mRNA and protein secretion of TF, TFPI, PAI-1, but not t-PA, was increased in trophoblast cell culture under hypoxia and hypoxia-reoxygenation. Cell cultures with high levels of tumor necrosis factor-alpha (TNF-alpha) exhibited increased expression and secretion of TF and PAI-1, decreased TFPI, and no significant change of t-PA. CONCLUSIONS: Imbalanced synthesis of t-PA, PAI-1, TFPI, and TF in trophoblasts may contribute to hypercoagulability in patients with preeclampsia. PMID- 19290853 TI - Progesterone - induced blocking factor (PIBF) and Th(1)/Th(2) cytokine in women with threatened spontaneous abortion. AB - OBJECTIVE: The aim of this prospective study was to compare serum and urine concentrations of progesterone-induced blocking factor (PIBF) and serum concentrations of anti-inflammatory (IL10) and pro-inflammatory (IL6, TNFalpha, IFNgamma) cytokines of women with threatened spontaneous abortion with normal pregnancy and to evaluate the impact of PIBF on outcome of pregnancy. METHODS: A sample of 30 women with threatened spontaneous abortion (study group) and 20 healthy pregnant women (control group) between 6(th) and 24(th) gestational weeks was studied. Serum and urine PIBF, IL10 and IL6, TNFalpha, IFNgamma cytokine concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Five (16.7%) pregnancies in the study group ended missed abortion vs. none in the control group (P<0.05). Five (20%) threatened aborters delivered between 24(th) and 37(th) weeks of gestation, whereas two (10%) preterm deliveries occurred in the controls (P>0.05). PIBF concentrations in urine (19.5+/-12.9 ng/mL) and serum (214.4+/-120.6 of patients with threatened abortion were significantly lower than in healthy pregnant women (45.3+/-33.7 ng/mL and 357.3+/-159.9 ng/mL, respectively). Women with threatened abortion had significantly lower serum levels of anti-inflammatory cytokine, but levels of proinflammatory cytokines were higher in this group compared with healthy controls. CONCLUSIONS: Determination of progesteron-induced blocking factor level in body fluids in early pregnancy might be used for the diagnosis and prognosis of threatened abortion. PMID- 19290854 TI - Parental mental representations during late pregnancy and early parenthood following assisted reproductive technology. AB - OBJECTIVE: To evaluate the relationship between assisted reproduction technology (ART) and parental mental representations during late pregnancy and early parenthood. STUDY DESIGN: Women (n=25) following ART pregnancies were compared with their partners (n=23) and with women following spontaneous conceptions (n=39). Subjects were interviewed on mental representations at late gestation and three months postpartum. RESULTS: Socio-demographic and obstetrical characteristics were similar between groups. ART women tend to decrease scores of intensity of investment from before to after delivery. During pregnancy and postpartum, ambivalent representations were more often present and well integrated representations were less often present among ART women as compared with non-ART women. ART women had significantly more ambivalent representations that persisted at three months postpartum, and men had more disengaged representations. CONCLUSIONS: A greater desire for pregnancy exists in ART women which might not always coincide with a desire for maternity. ART men, however, manifest greater disengagement from the entire child project. PMID- 19290855 TI - Flowcytometric assessment of fetomaternal hemorrhage during external cephalic version at term. AB - External cephalic version (ECV) at term is a safe procedure and reduces the incidence of cesarean sections for breech presentation. One of the known complications, however, is an ECV-related disruption of the placental barrier and a subsequent transfusion of fetal blood into maternal circulation. While the incidence of ECV-related fetomaternal hemorrhage (FMH) has been determined recently in a large trial using a manual Kleihauer-Betke test (KBT), questions remain on the amount of ECV-related FMH. KBT, which detects fetal red blood cells (RBC) on the basis of acidic resistance of fetal hemoglobin (HbF), is known to be a sensitive test, yet prone to procedural errors limiting its accuracy in quantifying FMH. In this study we investigated 50 patients for FMH before and after ECV, using a dual-color flow cytometric test kit with a lower limit of quantification of 0.05% fetal RBC in maternal peripheral blood. Three patients had a quantifiable increase of fetal RBC detected after ECV (0.06%; 0.08%; 0.1%). None of these subtle increments was predictable by ECV-related clinical parameters or translated into fetal compromise. Using a sensitive and accurate flow cytometric test method, our data provide further assurance to mothers on the safety of ECV at term. PMID- 19290856 TI - Does progesterone inhibit bacteria-stimulated interleukin-8 production by lower genital tract epithelial cells? AB - OBJECTIVE: Progesterone (P(4)) has been clinically shown to prevent the recurrence of preterm birth. The mechanism(s) of action is unclear, but may involve modulation of the immunologic inflammatory response of the lower genital tract. We evaluated the effects of P(4) on interleukin-8 (IL-8) production by vaginal and cervical epithelial cells stimulated with bacterial species that are commonly associated with preterm birth. METHODS: Vaginal and endocervical epithelial cells were incubated with up to 10,000 ng/mL P(4) overnight and stimulated with heat-killed Escherichia coli, Gardnerella vaginalis, or Ureaplasma urealyticum. Concentrations of IL-8 in conditioned medium were quantified by ELISA and viability of the cell cultures was measured by the reduction of a tetrazolium salt. RESULTS: E. coli, G. vaginalis and U. urealyticum-stimulated IL-8 production for both cell lines. P(4) inhibited basal and bacteria-stimulated IL-8 production for vaginal epithelial cells but enhanced IL-8 production by endocervical cells. P(4) reduced the number of viable cells for both cell lines. CONCLUSIONS: P(4) inhibits IL-8 production by vaginal epithelial cells stimulated with pathogens associated with preterm birth, possibly by reducing the number of viable cells or by inhibiting their proliferation. Although P(4) also reduces proliferation of endocervical cells it also increases their production of IL-8. PMID- 19290857 TI - Enzyme mechanisms: fast reaction and computational approaches. AB - Now, more than ever, enzymology and its development can be considered of vital importance to the progression of the biological sciences. With an increase in the numbers of enzymes being identified from genomic studies, enzymology is key to defining the structural and functional properties of these enzymes in order to establish their mechanisms of action and how they fit into metabolic networks. Along with the efforts of the bioinformaticians and systems biologists, such studies will ultimately lead to detailed descriptions of intricate biochemical pathways and allow identification of the most appropriate target enzymes for intervention in disease therapy. Thus the timing for the recent Biochemical Society Focused Meeting entitled 'Enzyme Mechanisms: Fast Reaction and Computational Approaches' was highly appropriate. The present paper represents an overview of this meeting, which was held at the Manchester Interdisciplinary Biocentre on 9-10 October 2008. PMID- 19290858 TI - Insights into the mechanisms of adenosylcobalamin (coenzyme B12)-dependent enzymes from rapid chemical quench experiments. AB - Glutamate mutase is one of a group of adenosylcobalamin-dependent enzymes that use free radicals to catalyse unusual and chemically difficult rearrangements involving 1,2-migrations of hydrogen atoms. A key mechanistic feature of these enzymes is the transfer of the migrating hydrogen atom between substrate, coenzyme and product. The present review summarizes recent experiments from my laboratory that have used rapid chemical quench techniques to identify intermediates in the reaction and probe the mechanism of hydrogen transfer through a variety of pre-steady-state kinetic isotope effect measurements. PMID- 19290859 TI - An end to 40 years of mistakes in DNA-protein association kinetics? AB - Proteins that bind to specific sequences in long DNA molecules have to locate their target sites amid myriad alternative sequences, yet they do so at remarkably rapid rates, sometimes approaching 10(10) M(-1) x s(-1). Hence, it has been asserted widely that binding to specific DNA sites can surpass the maximal rate for 3D (three-dimensional) diffusion through solution and that this could only be accounted for by a reduction in the dimensionality of the search for the target in effect by 1D (one-dimensional) diffusion (or 'sliding') along the DNA contour. It will be shown here that there is, in fact, no known example of a protein binding to a specific DNA site at a rate above the diffusion limit, and that the rapidity of these reactions is due primarily to electrostatic interactions between oppositely charged molecules. It will also be shown that, contrary to popular belief, reduced dimensionality does not, in general, increase the rate of target-site location but instead reduces it. Finally, it will be demonstrated that proteins locate their target sites primarily by multiple dissociation/reassociation events to other (nearby or distant) sites within the same DNA molecule, and that 1D diffusion is limited to local searches covering approximately 50 bp around each landing site. PMID- 19290860 TI - Probing coupled motions in enzymatic hydrogen tunnelling reactions. AB - Much work has gone into understanding the physical basis of the enormous catalytic power of enzymes over the last 50 years or so. Nevertheless, the detailed mechanism used by Nature's catalysts to speed chemical transformations remains elusive. DHFR (dihydrofolate reductase) has served as a paradigm to study the relationship between the structure, function and dynamics of enzymatic transformations. A complex reaction cascade, which involves rearrangements and movements of loops and domains of the enzyme, is used to orientate cofactor and substrate in a reactive configuration from which hydride is transferred by quantum mechanical tunnelling. In the present paper, we review results from experiments that probe the influence of protein dynamics on the chemical step of the reaction catalysed by TmDHFR (DHFR from Thermotoga maritima). This enzyme appears to have evolved an optimal structure that can maintain a catalytically competent conformation under extreme conditions. PMID- 19290861 TI - Conformational changes in the catalytic cycle of protochlorophyllide oxidoreductase: what lessons can be learnt from dihydrofolate reductase? AB - In chlorophyll biosynthesis, the light-activated enzyme, POR (protochlorophyllide oxidoreductase), has been shown to be an excellent model system for studying the role of protein motions during catalysis. The catalytic cycle of POR is understood in detail and comprises an initial photochemical reaction, which is followed by a number of 'dark' steps. The latter steps in the reaction cycle have been shown to involve a series of ordered product release and substrate rebinding events and are known to require conformational changes in the protein in order to proceed. However, owing to the current lack of any structural information on the enzyme, the nature of these conformational rearrangements remains poorly understood. By contrast, there is a wealth of structural and kinetic information available on the closely related enzyme dihydrofolate reductase, which is known to have a similar catalytic mechanism to POR. Dihydrofolate reductase is able to adopt an 'occluded' and a 'closed' structure, depending on which ligand is bound in the active site, and as a result, the catalytic cycle is controlled by a 'switching' between these two conformations. By analogy, we suggest that a similar cycling between different conformations may be operating in POR. PMID- 19290862 TI - Time-resolved studies of radical pairs. AB - The effect of magnetic fields on chemical reactions through the RP (radical pair) mechanism is well established, but there are few examples, in the literature, of biological reactions that proceed through RP intermediates and show magnetic field-sensitivity. The present and future relevance of magnetic field effects in biological reactions is discussed. PMID- 19290863 TI - Insights into the mechanism and inhibition of fatty acid amide hydrolase from quantum mechanics/molecular mechanics (QM/MM) modelling. AB - FAAH (fatty acid amide hydrolase) is a promising target for the treatment of several central nervous system and peripheral disorders. Combined QM/MM (quantum mechanics/molecular mechanics) calculations have elucidated the role of its unusual catalytic triad in the hydrolysis of oleamide and oleoylmethyl ester substrates, and have identified the productive inhibitor-binding orientation for the carbamoylating compound URB524. These are potentially crucial insights for designing new covalent inhibitors of this drug target. PMID- 19290864 TI - Opportunities for mesoporous nanocrystalline SnO2 electrodes in kinetic and catalytic analyses of redox proteins. AB - PFV (protein film voltammetry) allows kinetic analysis of redox and coupled chemical events. However, the voltammograms report on the electron transfer through a flow of electrical current such that simultaneous spectroscopy is required for chemical insights into the species involved. Mesoporous nanocrystalline SnO(2) electrodes provide opportunities for such 'spectroelectrochemical' analyses through their high surface area and optical transparency at visible wavelengths. Here, we illustrate kinetic and mechanistic insights that may be afforded by working with such electrodes through studies of Escherichia coli NrfA, a pentahaem cytochrome with nitrite and nitric oxide reductase activities. In addition, we demonstrate that the ability to characterize electrocatalytically active protein films by MCD (magnetic circular dichroism) spectroscopy is an advance that should ultimately assist our efforts to resolve catalytic intermediates in many redox enzymes. PMID- 19290865 TI - Density functional theory (DFT) and combined quantum mechanical/molecular mechanics (QM/MM) studies on the oxygen activation step in nitric oxide synthase enzymes. AB - In this review paper, we will give an overview of recent theoretical studies on the catalytic cycle(s) of NOS (nitric oxide synthase) enzymes and in particular on the later stages of these cycles where experimental work is difficult due to the short lifetime of intermediates. NOS enzymes are vital for human health and are involved in the biosynthesis of toxic nitric oxide. Despite many experimental efforts in the field, the catalytic cycle of this important enzyme is still surrounded by many unknowns and controversies. Our theoretical studies were focused on the grey zones of the catalytic cycle, where intermediates are short lived and experimental detection is impossible. Thus combined QM/MM (quantum mechanics/molecular mechanics) as well as DFT (density functional theory) studies on NOS enzymes and active site models have established a novel mechanism of oxygen activation and the conversion of L-arginine into N(omega)-hydroxo arginine. Although NOS enzymes show many structural similarities to cytochrome P450 enzymes, it has long been anticipated that therefore they should have a similar catalytic cycle where molecular oxygen binds to a haem centre and is converted into an Fe(IV)-oxo haem(+*) active species (Compound I). Compound I, however, is elusive in the cytochrome P450s as well as in NOS enzymes, but indirect experimental evidence on cytochrome P450 systems combined with theoretical modelling have shown it to be the oxidant responsible for hydroxylation reactions in cytochrome P450 enzymes. By contrast, in the first catalytic cycle of NOS it has been shown that Compound I is first reduced to Compound II before the hydroxylation of arginine. Furthermore, substrate arginine in NOS enzymes appears to have a dual function, namely first as a proton donor in the catalytic cycle to convert the ferric-superoxo into a ferric-hydroperoxo complex and secondly as the substrate that is hydroxylated in the process leading to N(omega)-hydroxo-arginine. PMID- 19290866 TI - Combining X-ray crystallography and single-crystal spectroscopy to probe enzyme mechanisms. AB - The combination of X-ray crystallography and rapid cryo-trapping methods has enabled the visualization of catalytic intermediates in a variety of enzyme systems. However, the resolution of the X-ray experiment is not always sufficient to precisely place the structure on the reaction pathway. In addition, many trapped intermediates are X-ray-sensitive and can decay during diffraction data collection, resulting in a final structure that may not be representative of the initial trapped species. Complementary methods, such as single-crystal spectroscopy, provide a means to precisely identify the cryo-trapped species as well as detect any X-ray-induced changes during diffraction data collection. PMID- 19290867 TI - Tryptophan 13C nuclear-spin polarization generated by intraprotein electron transfer in a LOV2 domain of the blue-light receptor phototropin. AB - (13)C-NMR experiments were performed on photo-excited fully and partially (13)C labelled LOV2 domains of the blue-light receptor phototropin. In the present paper, we report on nuclear-spin polarized tryptophan resonances that are generated by light-induced intraprotein electron transfer to the FMN cofactor. The spectra are discussed with respect to earlier data obtained from (13)C-NMR experiments on unlabelled LOV2 domains that have been reconstituted with FMN (13)C isotopologues. PMID- 19290868 TI - Ultrafast catalytic processes and conformational changes in the light-driven enzyme protochlorophyllide oxidoreductase (POR). AB - The enzyme POR (protochlorophyllide oxidoreductase), from the family of alcohol dehydrogenases, reduces protochlorophyllide into chlorophyllide on the absorption of light. The reduction involves the transfer of two protons and two electrons and is an important regulatory step in the biosynthesis of chlorophyll. In recent years, due to the availability of large quantities of the pure enzyme, much of the catalytic reaction has been unravelled by using a variety of spectroscopic methods, including ultrafast initial events in catalysis. In addition, it has been demonstrated that a light-activated conformational change of the protein is necessary to activate catalysis. This makes POR a very important model system to study the relationship between structural changes of enzymes and functionality. PMID- 19290869 TI - The bacterial respiratory nitric oxide reductase. AB - The two-subunit cytochrome bc complex (NorBC) isolated from membranes of the model denitrifying soil bacterium Paracoccus denitrificans is the best characterized example of the bacterial respiratory nitric oxide reductases. These are members of the super-family of haem-copper oxidases and are characterized by the elemental composition of their active site, which contains non-haem iron rather than copper, at which the reductive coupling of two molecules of nitric oxide to form nitrous oxide is catalysed. The reaction requires the presence of two substrate molecules at the active site along with the controlled input of two electrons and two protons from the same side of the membrane. In the present paper, we consider progress towards understanding the pathways of electron and proton transfer in NOR and how this information can be integrated with evidence for the likely modes of substrate binding at the active site to propose a revised and experimentally testable reaction mechanism. PMID- 19290870 TI - The petite purple photosynthetic powerpack. AB - Photoreaction centres are Nature's solar batteries. These nanometre-scale power producers are responsible for transducing the energy of sunlight into a form that can be used by biological systems, thereby powering most of the biological activity on the planet. Although to the layman the word 'photosynthesis' is usually associated with green plants, much of our understanding of the molecular basis of biological transduction of light energy has come from studies of purple photosynthetic bacteria. Their RCs (reaction centres) and attendant light harvesting complexes have been subjected to an intensive spectroscopic scrutiny, coupled with genetic manipulation and structural studies, that has revealed many of the molecular and mechanistic details of biological energy transfer, electron transfer and coupled proton translocation. This review provides a short overview of the structure and mechanism of the purple bacterial RC, focusing in the main on the most heavily studied complex from Rhodobacter sphaeroides. PMID- 19290871 TI - Oxidation of L-tryptophan in biology: a comparison between tryptophan 2,3 dioxygenase and indoleamine 2,3-dioxygenase. AB - The family of haem dioxygenases catalyse the initial oxidative cleavage of L tryptophan to N-formylkynurenine, which is the first, rate-limiting, step in the L-kynurenine pathway. In the present paper, we discuss and compare structure and function across the family of haem dioxygenases by focusing on TDO (tryptophan 2,3-dioxygenase) and IDO (indoleamine 2,3-dioxygenase), including a review of recent structural information for both enzymes. The present paper describes how the recent development of recombinant expression systems has informed our more detailed understanding of the substrate binding, catalytic activity and mechanistic properties of these haem dioxygenases. PMID- 19290872 TI - Mechanism of CB1954 reduction by Escherichia coli nitroreductase. AB - NTR (nitroreductase NfsB from Escherichia coli) is a flavoprotein with broad substrate specificity, reducing nitroaromatics and quinones using either NADPH or NADH. One of its substrates is the prodrug CB1954 (5-[aziridin-1-yl]-2,4 dinitrobenzamide), which is converted into a cytotoxic agent; so NTR/CB1954 has potential for use in cancer gene therapy. However, wild-type NTR has poor kinetics and binding with CB1954, and the mechanism for the reduction of CB1954 by NTR is poorly understood. Computational methods have been utilized to study potential underlying reaction mechanisms so as to identify the order of electron and proton transfers that make up the initial reduction step and the sources of the protons. We have used Molecular Dynamics to examine the nature of the active site of the wild-type enzyme and the preferred binding mode of the substrate. A combination of these results has allowed us to unequivocally identify the reaction mechanism for the reduction of CB1954 by NTR. PMID- 19290873 TI - Nucleic acid detection and quantification in the developing world. AB - Techniques using nucleic acid amplification have not had the same amount of impact on research and clinical diagnosis in the developing world as that observed in the West. This is unsurprising when the costs and infrastructure required to perform nucleic acid amplification are considered. Despite this, nucleic acid amplification is being increasingly used in both research and diagnosis in countries such as Zambia and Tanzania. Scientific research in the developing world is made possible through the support and development of the necessary laboratory infrastructure and the establishment of special transport for the reagents and samples. This has enabled world-leading country-relevant research to be performed by local scientists on subjects ranging from rapid diagnosis of infectious diseases to measuring the RNA gene expression in an immune response. Concomitantly, the challenge presented by the need for tests that are more appropriate for a resource-poor setting has led to a number of newer methodologies for nucleic acid detection, which can be tailored to be performed in the field without the need for training in molecular biology. As nucleic acid amplification techniques become both simpler and cheaper, their impact is likely to play an increasingly crucial role in research and diagnosis in the developing world. PMID- 19290874 TI - Miniaturized PCR systems for cancer diagnosis. AB - PCR retains a pivotal role in making accessible marker nucleic acid sequences for ready analysis in cancer diagnosis. For certain cancers such as acute lymphoblastic leukaemia, the application of quantitative procedures to assess and subsequently direct therapy has given rise to the slowly maturing field of MRD (minimal residual disease) management. Although excellent protocols exist for performing these analyses, akin to all PCR procedures the limit of detection can vary markedly between laboratories. The present paper is an overview that describes how the analytical field relating to miniaturization is likely to identify the missing link that integrates sample processing with downstream PCR, analysis and eventual therapy. Miniaturized devices are suited to the multi parallelized handling of defined numbers of cells, and PCR-based microfluidic procedures have become reasonably established. The integration of sample processing and PCR in microfluidic devices is beginning to offer reproducible quantitative data that relate the number of biomarker nucleic acids to the defined analysed cell or cells for meaningful clinical assessment. The application of MRD may, through integrated miniaturized PCR, become more reliable and routine with additional applications in defining disease threshold levels for other cancer types. These enabling integrated platforms may facilitate biomarker measurements to predict the response and outcome, which are also of current interest for personalized medical care. PMID- 19290875 TI - COLD-PCR: a new platform for highly improved mutation detection in cancer and genetic testing. AB - PCR is widely employed as the initial DNA amplification step for genetic testing and cancer biomarker detection. However, a key limitation of PCR-based methods, including real-time PCR, is the inability to selectively amplify low levels of variant alleles in a wild-type allele background. As a result, downstream assays are limited in their ability to identify subtle genetic changes that can have a profound impact on clinical decision-making and outcome or that can serve as cancer biomarkers. We developed COLD-PCR (co-amplification at lower denaturation temperature-PCR) [Li, Wang, Mamon, Kulke, Berbeco and Makrigiorgos (2008) Nat. Med. 14, 579-584], a novel form of PCR that amplifies minority alleles selectively from mixtures of wild-type and mutation-containing sequences irrespective of the mutation type or position on the sequence. Consequently, COLD PCR amplification from genomic DNA yields PCR products containing high-prevalence variant alleles that can be detected. Since PCR constitutes a ubiquitous initial step for almost all genetic analysis, COLD-PCR provides a general platform to improve the sensitivity of essentially all DNA-variation detection technologies including Sanger sequencing, pyrosequencing, single molecule sequencing, mutation scanning, mutation genotyping or methylation assays. COLD-PCR combined with real time PCR provides a new approach to boost the capabilities of existing real-time mutation detection methods. We replaced regular PCR with COLD-PCR before sequencing or real-time mutation detection assays to improve mutation detection sensitivity by up to 100-fold and identified novel p53/Kras/EGFR (epidermal growth factor receptor) mutations in heterogeneous cancer samples that were missed by all existing methods. For clinically relevant micro-deletions, COLD-PCR enabled exclusive amplification and isolation of the mutants. COLD-PCR is expected to have diverse applications in the fields of biomarker identification and tracing, genomic instability, infectious diseases, DNA methylation testing and prenatal identification of fetal alleles in maternal blood. PMID- 19290876 TI - Mutation scanning using high-resolution melting. AB - Mutation scanning techniques are used to detect sequence variants without the need for prior knowledge of the identity or precise location of the variant, in contrast with genotyping techniques, which determine the status of a specific variant. High-resolution melting is a recently developed method that shows great potential as a mutation scanning technique. Sensitivity and specificity for mutation detection are extremely high and the technique also has advantages of cost and throughput. Practical considerations for successful mutation scanning by high-resolution melting are also discussed in this review. PMID- 19290877 TI - PCR in forensic genetics. AB - Since the introduction in the mid-1980s of analyses of minisatellites for DNA analyses, a revolution has taken place in forensic genetics. The subsequent invention of the PCR made it possible to develop forensic genetics tools that allow both very informative routine investigations and still more and more advanced, special investigations in cases concerning crime, paternity, relationship, disaster victim identification etc. The present review gives an update on the use of DNA investigations in forensic genetics. PMID- 19290878 TI - Functionalized nanoparticles for nucleic acid sequence analysis using optical spectroscopies. AB - SERRS (surface-enhanced resonance Raman scattering) is a vibrational spectroscopy which allows extremely sensitive and selective detection of labelled DNA sequences with detection limits which rival, and in most cases surpass, that of fluorescence. SERRS relies on a visible chromophore adsorbing on to an enhancing surface. DNA itself is not SERRS-active, as it lacks a suitable visible chromophore and has poor adsorption properties on to the surfaces used for enhancement. The surface normally used for enhancement in these sorts of studies are metallic nanoparticles and, through modification of DNA probes by the addition of suitable SERRS labels, signals can be obtained that are highly sensitive and very selective. The aggregation state of the nanoparticles is critical to the sensitivity, and, in the present paper, we show how straightforward detection of labelled DNA probes can be achieved using SERRS in a quantitative manner and with a variety of different commercially available labels. In a second approach, we show how the properties of aggregation to turn on the SERRS effect can be exploited through DNA hybridization to give identification of a particular DNA sequence. This approach lends itself to closed tube formats and is a promising way forward for molecular diagnostics using SERRS. PMID- 19290879 TI - Optical fibre-based detection of DNA hybridization. AB - A dual-peak LPFG (long-period fibre grating), inscribed in an optical fibre, has been employed to sense DNA hybridization in real time, over a 1 h period. One strand of the DNA was immobilized on the fibre, while the other was free in solution. After hybridization, the fibre was stripped and repeated detection of hybridization was achieved, so demonstrating reusability of the device. Neither strand of DNA was fluorescently or otherwise labelled. The present paper will provide an overview of our early-stage experimental data and methodology, examine the potential of fibre gratings for use as biosensors to monitor both nucleic acid and other biomolecular interactions and then give a summary of the theory and fabrication of fibre gratings from a biological standpoint. Finally, the potential of improving signal strength and possible future directions of fibre grating biosensors will be addressed. PMID- 19290880 TI - Genomic DNA amplification by the multiple displacement amplification (MDA) method. AB - Large amounts of DNA are frequently required for use in detection assays and genomic analysis. The limited availability of DNA can be a critical obstacle to meeting research and clinical needs. DNA amplification methods are often required to generate sufficient material from small specimens or environmental samples with low DNA content. The MDA (multiple displacement amplification) reaction is increasingly the method of choice for many applications because of its extensive coverage of the genome, the generation of extremely long DNA products compared with older whole genome amplification methods and the high DNA yields, even from exceedingly low amounts of starting material. Remarkably, MDA enables genomic sequencing even from single microbial cells. Some of the uses of MDA and its strengths and limitations will be discussed. PMID- 19290881 TI - The single-nucleotide primer extension (SNuPE) method for the multiplex detection of various DNA sequences: from detection of point mutations to microbial ecology. AB - Methods based on SNuPE (single-nucleotide primer extension) have become invaluable tools for the rapid and highly specific detection of point mutations and single-nucleotide polymorphisms in the field of human genetics. In the primer extension reaction, a DNA polymerase is used to label a specific primer hybridized to the target sequence by incorporating a single labelled ddNTP (dideoxynucleotide). This labelling provides not only information about the complementary nucleotide of interest in the opposite strand but also a semiquantitative analysis of the sequence targeted by the primer. Since several subdisciplines of microbiology increasingly require cultivation-independent molecular screening tools for elucidating differences between either strains or community structures based on sequence variations of marker genes, SNuPE offers a promising alternative to the existing tool box. The present review describes the method in detail and reports the state-of-the-art applications of this technique both in the field of nucleic acid detections in human genetics and in microbiology. PMID- 19290882 TI - The SAFE project: towards non-invasive prenatal diagnosis. AB - After the revolutionary detection of ffDNA (free fetal DNA) in maternal circulation by real-time PCR in 1997 and advances in molecular techniques, NIPD (non-invasive prenatal diagnosis) is now a clinical reality. Non-invasive diagnosis using ffDNA has been implemented, allowing the detection of paternally inherited alleles, sex-linked conditions and some single-gene disorders and is a viable indicator of predisposition to certain obstetric complications [e.g. PET (pre-eclampsia)]. To date, the major use of ffDNA genotyping in the clinic has been for the non-invasive detection of the pregnancies that are at risk of HDFN (haemolytic disease of the fetus and newborn). This has seen numerous clinical services arising across Europe and many large-scale NIPD genotyping studies taking place using maternal plasma. Because of the interest in performing NIPD and the speed at which the research in this area was developing, the SAFE (Special Non-Invasive Advances in Fetal and Neonatal Evaluation) NoE (Network of Excellence) was founded. The SAFE project was set up to implement routine, cost effective NIPD and neonatal screening through the creation of long-term partnerships within and beyond the European Community and has played a major role in the standardization of non-invasive RHD genotyping. Other research using ffDNA has focused on the amount of ffDNA present in the maternal circulation, with a view to pre-empting various complications of pregnancy. One of the key areas of interest in the non-invasive arena is the prenatal detection of aneuploid pregnancies, particularly Down's syndrome. Owing to the high maternal DNA background, detection of ffDNA from maternal plasma is very difficult; consequently, research in this area is now more focused on ffRNA to produce new biomarkers. PMID- 19290883 TI - Dissolved oxygen alteration of the spectrophotometric analysis and quantification of nucleic acid solutions. AB - Nucleic acids are routinely and readily analysed using the A(260)/A(280) ratio, although this method is known to be prone to erroneous results owing to contaminants in solution that absorb at similar wavelengths. The aim of the present review, while highlighting the problems and alternatives of using UV spectrophotometry for nucleic acid measurements, is to bring forth an observational result from our recent studies, namely that DO (dissolved oxygen) and nitrogen can alter the A(260) of aqueous DNA solutions. Our finding is of importance because DO is highly variable between protocols and storage conditions of DNA preparations. The physicochemical nature of the oxygen-DNA interactions is briefly discussed. PMID- 19290884 TI - Real-time DNA microarrays: reality check. AB - DNA microarrays are plagued with inconsistent quantifications and false-positive results. Using established mechanisms of surface reactions, we argue that these problems are inherent to the current technology. In particular, the problem of multiplex non-equilibrium reactions cannot be resolved within the framework of the existing paradigm. We discuss the advantages and limitations of changing the paradigm to real-time data acquisition similar to real-time PCR methodology. Our analysis suggests that the fundamental problem of multiplex reactions is not resolved by the real-time approach itself. However, by introducing new detection chemistries and analysis approaches, it is possible to extract target-specific quantitative information from real-time microarray data. The possible scope of applications for real-time microarrays is discussed. PMID- 19290886 TI - Ca2+/H+ antiporter-like activity of human recombinant Bax inhibitor-1 reconstituted into liposomes. AB - We investigated the functional activity of recombinant Bax inhibitor-1 reconstituted into liposomes. When proteoliposomes were suspended in acidic solutions, encapsulated Ca(2+) was released from the membranes, as previously suggested [Kim HR, Lee GH, Ha KC, Ahn T, Moon JY, Lee BJ, Cho SG, Kim S, Seo YR, Shin YJ et al. (2008) J Biol Chem283, 15946-15955]. Concomitantly, proton ions were internalized when assayed using the time-dependent change in the fluorescence of the pH-sensitive dye oxonol V entrapped in the proteoliposomes. The influx of proton ions was confirmed by observing tritium accumulation in the membranes. However, the external acidity of the membranes per se did not induce proton ion influx without internalized Ca(2+). These results suggest that reconstituted Bax inhibitor-1 has a Ca(2+)/H(+) antiporter-like activity. PMID- 19290887 TI - Full-length adiponectin protects hepatocytes from palmitate-induced apoptosis via inhibition of c-Jun NH2 terminal kinase. AB - Hepatic apoptosis is elevated in patients with non-alcoholic steatohepatitis and is correlated with the severity of the disease. Long-chain saturated fatty acids, such as palmitate, induce apoptosis in liver cells. The present study examined adiponectin-mediated protection against saturated fatty acid-induced apoptosis in the human hepatoma cell line, HepG2. Cells were cultured in a control media (i.e. without fatty acids) or the same media containing 250 micromol L(-1) of albumin bound oleate or palmitate for 24 h. The adiponectin concentrations used were: 0, 1, 10 or 100 microg mL(-1) (n = 4-6 per treatment). Palmitate and thapsigargin, but not oleate, activated caspase-3 and decreased cell viability in the absence of adiponectin. Adiponectin reduced palmitate- and thapsigargin-induced activation of caspase-3 and cell death in a dose-dependent manner. Phosphatidylinositol 3-kinase and AMP-activated protein kinase inhibitors abolished the effects of adiponectin. Adiponectin-induced inhibition of palmitate and thapsigargin-induced apoptosis was not the result of an augmentation in the unfolded protein response or the increased expression of genes encoding the inhibitor of apoptosis proteins, inhibitor of apoptosis protein-2 and X-linked mammalian inhibitor of apoptosis protein. Palmitate and thapsigargin, but not oleate, increased c-Jun NH(2) terminal kinase phosphorylation in the absence of adiponectin. Adiponectin blocked palmitate- and thapsigargin-induced activation of c-Jun NH(2) terminal kinase and reduced apoptosis. These data suggest that adiponectin is an important determinant of saturated fatty acid-induced apoptosis in liver cells and may have implications for fatty acid-mediated liver cell injury in adiponectin-deficient individuals. PMID- 19290888 TI - Integrated treatment to resolve a horizontal root fracture. AB - Root fractures are defined as those that involve cement, dentin and pulp, comprising from 0.5 to 7% of injuries in permanent dentition. Diagnosis is made through clinical and radiographic exams, the latter frequently being limited by the position of the fracture. Treatment varies according to the displacement and vitality of the fragments. The authors present a clinical case of recurrent trauma of tooth 21 causing a horizontal root fracture in the middle third. After several attempts at endodontic treatment, the option was to remove the apical fragment by surgery. The postoperative period of 4 years shows very satisfactory results with regard to wound repair and tooth mobility, or implantation of the coronal segment. PMID- 19290889 TI - Parasymphyseal fracture associated with fracture of a maxillary primary molar in a child: case report. AB - Bone and posterior tooth fractures can be serious injuries that are difficult to diagnose, particularly in emergencies. The aim of this study was to report a case of a parasymphyseal fracture associated with the fracture of a maxillary primary molar in a child. PMID- 19290890 TI - Mesiodens as a risk factor in treatment of trauma cases. AB - The presence of mesiodens often results in complications including retention of primary teeth and delayed eruption of permanent teeth, closure of eruption path, rotations, retention, root resorption, pulp necrosis, and diastema as well as nasal eruption and formation of dentigerous and primordial cysts. Less common complications involving the permanent incisors include dilaceration of the developing roots and loss of tooth vitality. Therefore, early diagnosis of mesiodens has particular importance in terms of preventing such complications. However, with respect to surgical removal of mesiodens, ideal timing of intervention - immediate or delayed intervention - remains to be a highly controversial issue. While predisposing factors of dental trauma such as open bite, increasing overjet with protrusion of upper incisors, and insufficient lip closure have been well documented in the literature, there is limited evidence indicating mesiodens as a risk factor in trauma. In this report of two cases, it is aimed to emphasize mesiodens as a risk factor in the treatment of dental trauma besides its effect on prognosis. PMID- 19290891 TI - The long road to evidence-based traumatology. PMID- 19290892 TI - Splinting duration and periodontal outcomes for replanted avulsed teeth: a systematic review. AB - Clinical guidelines are now available for the management of avulsed teeth. The principles of evidence-based dentistry can be used to assess whether these guidelines are based on currently-available evidence. A qualitative systematic review was conducted of relevant clinical literature to examine the evidence on splinting duration and periodontal healing outcomes. The review was constrained markedly by small sample sizes, retrospective nature of clinical audits, dissimilarities of selected studies in their design, methodology and observation periods, and lack of uniformity in terminology for outcomes. A total of 138 replanted avulsed permanent teeth pooled from four papers each reporting both short-term splinting (14 days or less) and long-term splinting (over 14 days) in accord with current clinical guidelines, were studied. The evidence for an association between short-term splinting and an increased likelihood of functional periodontal healing, acceptable healing, or decreased development of replacement resorption, appears inconclusive. The study found no evidence to contraindicate the current guidelines and suggests that the likelihood of successful periodontal healing after replantation is unaffected by splinting duration. Pending future research to the contrary, it is recommended that dentists continue to use the currently-recommended splinting periods when replanting avulsed permanent teeth. PMID- 19290893 TI - An evidence-based assessment of the clinical guidelines for replanted avulsed teeth. Part II: prescription of systemic antibiotics. AB - BACKGROUND: Current clinical guidelines recommend prescribing systemic antibiotic therapy (SAT) for patients having an avulsed permanent tooth replanted. The principles of evidence-based dentistry can be used to assess whether this is the best approach based on currently-available evidence. The objective of this study was to use the principles of evidence-based dentistry to answer the PICO question: (P) for a replanted avulsed permanent tooth, (I) is prescribing SAT, (C) compared with not prescribing SAT, (O) associated with an increased likelihood of successful periodontal healing after tooth replantation? MATERIALS AND METHODS: A literature search was performed across four internet databases (Ovid Medline, Cochrane Library, PubMed, ISI Web of Science), for relevant citations (n = 35 702). Limiting citations to those in English and removing duplicates produced a set of titles (n = 14 742) that were sieved according to evidence-based dentistry principles. Relevant titles were selected for abstract assessment (n = 782), identifying papers for examination (n = 74). Inclusion criteria were applied and three papers (326 total teeth) met the final criteria for meta-analysis. RESULTS: Meta-analyses found no statistically significant difference between prescribing or not prescribing antibiotics for acceptable periodontal healing without progressive root resorption (common odds ratio = 0.90, SE = 0.29, 95% confidence intervals = 0.51-1.58). CONCLUSION: The evidence for an association between prescribing SAT and an increased likelihood of acceptable periodontal healing outcome is inconclusive. This investigation of antibiotic use as defined in the clinical guidelines indicates there is inconclusive clinical evidence from studies of replanted avulsed human teeth to either contradict or support the guideline. Pending future research to the contrary, dentists are recommended to follow current guidelines in prescribing SAT when replanting avulsed teeth. PMID- 19290894 TI - Effects on permanent teeth after luxation injuries to the primary predecessors: a study in children assisted at an emergency service. AB - AIM: The purpose of this study is to evaluate the developmental disturbances in permanent teeth as a result of luxation injuries in the primary teeth predecessors. MATERIALS AND METHODS: A total of 5330 records, corresponding to 10 years attendance at the Emergency Center of Baby Clinic, Londrina State University, Brazil, were analyzed. Three hundred and eighty nine children were involved in this study, totaling 620 traumatized primary teeth. Clinical and radiographic examinations were carried out in the permanent successor teeth. RESULTS: In total, 623 permanent teeth were examined and developmental disturbances were detected in 126 teeth (20.2%). The white or yellow-brown discoloration of enamel was the clinical disturbance most observed (78.0%) and the crown alteration most detected through radiographic analysis was hypoplasia (86.0%). Root alterations were rare; root dilaceration was observed in only one case. The age of the children when their primary teeth received damage varied between 6 and 36 months (P = 0.000325). Intrusive luxation and avulsion were related with most of the cases of sequelae in the successor permanent teeth (P = 0.000001). CONCLUSION: The results of the present investigation emphasize the special attention required for children who suffered dental trauma at an early age, especially in cases of intrusive luxation and avulsion. PMID- 19290895 TI - Sequential surgical treatment for panfacial fractures and significance of biological osteosynthesis. AB - The goal of this retrospective study was to evaluate the efficacy of panfacial fracture repair and to review guidelines for treatment based on AO/Arbeitsgemeinschaft fuer Osteosynthesefragen Association for Study of Internal Fixation theories of biological osteosynthesis. Sixty-eight patients with panfacial fractures were subjected to preoperative X-ray cephalometric analysis and model surgery, followed by open surgical reduction, rigid internal fixation and at least 8 weeks of clinical follow up. A variety of surgical approaches were used, with the 68 patients undergoing a total of 93 surgical procedures. In all but eight patients, the treatment produced satisfactory correction of maxillofacial deformities and restoration of normal function. Among the eight patients whose treatment was not deemed successful, there were two whose facial deformities were not corrected by treatment. In addition, there were five patients with enophthalmos or motor disturbance of the eye that failed to show improvement during the study, and four patients who exhibited signs of limited mouth opening and malocclusion. The systematic and sequential choices of surgical methods were key factors in determining panfacial fracture treatment outcomes. We propose that treatment of bone fractures must take into account the biological characteristics of the damaged bone to facilitate selection of appropriate plate and screw systems and repositioning methods. PMID- 19290896 TI - Effect of different bleaching systems on the ultrastructure of bovine dentin. AB - The aim of this study was to evaluate in vitro the effect of different in-office bleaching systems on the surface morphology of bovine dentin. Thirty tooth fragments measuring 4 x 4mm, containing enamel and dentin, were obtained from the crowns of extracted bovine incisors. Samples were subjected to simulated intracoronal bleaching techniques using conventional (Opalescence Endo and Whiteness Super Endo) and light-activated systems (Opalescence Xtra) and Whiteness HP Maxx). Controls were treated with either sodium perborate mixed with 10% hydrogen peroxide or no bleaching agent. The samples were observed under SEM and the recorded images were evaluated for topographic alterations. The ultrastructural alterations of dentin observed in this study varied greatly between groups according to the products used. Higher pH products (Whiteness HP Maxx) and Opalescence Xtra) associated with in-office techniques yielded better maintenance of dentin ultrastructure. Apparently, both low pH and hydrogen peroxide oxidation play a role in altering the ultrastructure of dentin during internal dental bleaching. The use of alkaline products with reduced time of application (in-office techniques) may decrease such morphological alterations. PMID- 19290897 TI - Dental trauma in adults in Switzerland. AB - Most epidemiologic studies of dental trauma are restricted to children and sports. The goal of the present study was to collect information on dental trauma in as many Swiss adults as possible. For that purpose data of the largest insurance company of Switzerland (SUVA) were evaluated and compared for the years 1992 and 2002. In these 2 years 23 000 dental injuries were reported to all insurance companies in Switzerland. Most often the upper central incisors were injured. However, in 20% posterior teeth were also affected. In 1992 most of the dental trauma was experienced by 30-39 year olds, in 2002 by 40-49 year olds. Regardless of the year, most of the injuries occurred at home; more than one fifth of the injuries were sports related. Most common injuries were crown fractures without pulp involvement. In less than 10% of all cases more complicated injuries like avulsions or crown-root fractures occurred. Most traumatic injuries were treated by direct fillings and custom-made crowns; more involved treatments were required less often. Comparison of the years 1992 and 2002 showed that the numbers of and causes for injuries had hardly changed in these 10 years. However, the choice of therapy was related to the general development of dental medicine, which became especially apparent in the fields of adhesive dentistry and implantology. PMID- 19290898 TI - Dental trauma: an evidence-based approach to care. AB - BACKGROUND: One of the key aspects of improving the dental care we offer to our patients is in basing our practice on the best available evidence. AIM: The main purpose of this paper is to present some basic ways of searching reliable papers on Internet and to introduce some techniques to facilitate the development of strategic search skill to improve the quality of dental care, using dental trauma as an example. MATERIALS AND METHODS: After formulation of a clinical question on a dental trauma issue, a comparison was made between: data collected in PubMed using a given term; data collected in PubMed using the Medical Subject Headings vocabulary (Mesh) and data collected using the principles of evidence-based research, all by an independent researcher. A number of papers searched about a clinical question on dental trauma and a brief commentary about each Internet research database is presented. The authors reported that the best sites available to perform the sample search were those produced by academic centers, professional organizations and government-sponsored. To get evidence-based clinical papers we did internet search on PubMed, Cochrane, Center of Evidence Based Dentistry and Knowledge Finder using the clinical question "Emergency Treatment for Avulsed Tooth". RESULTS: All searched databases were efficient for scientific journals, but when we used the PI strategy, the search seemed to be more relevant and specific. The Cochrane, Center of Evidence-Based Dentistry and Knowledge Finder presented more reliable papers to answer our clinical question about dental trauma treatment. CONCLUSION: It is imperative that dentists understand the advantages of searching the Internet and learning to use it effectively to guide practice and assist their patients in their pursuit for better oral health. PMID- 19290899 TI - Maxillofacial fractures sustained during baseball and softball. AB - AIM: The purpose of this study was to investigate the demographics, the type of impact, the site and the treatment of maxillofacial fractures sustained during baseball and softball to develop an effective preventive strategy. PATIENTS AND METHODS: Data of 82 patients treated for baseball- and softball-related maxillofacial fractures at the Department of Oral and Maxillofacial Surgery, Nara Medical University between 1982 and 2007 were retrospectively analyzed. RESULTS: Injuries were found in 64 men in baseball and 16 men and two women in softball with average ages of 19.6 and 30.0 years, respectively. Fractures were caused by being hit by the ball in 61 patients followed by collision in 16 patients. Fractures of the mandible and the mid-face were found in 44 and 38 patients, respectively. The mental and angle region of the mandible and zygoma and alveolar bone of the maxilla were frequently involved. Treatment was mostly conservative. Open reduction and internal fixation were performed only in 15 patients. CONCLUSIONS: Most maxillofacial fractures in these sports were ball-related. Therefore, effective preventive means should be considered to protect against such injuries. PMID- 19290900 TI - Traumatic anterior dental injuries in 7- to 12-year-old Brazilian children. AB - The aim of this study was to determine the prevalence and risk factors of dental trauma in the permanent anterior teeth of schoolchildren in Campina Grande, Brazil. A sample of 448 schoolchildren, 228 boys and 220 girls, aged 7-12 years, were randomly selected from 17 public schools in an urban area. The sample selection was carried out in two stages: first, schools were selected by simple sampling and then children were chosen using a proportionality coefficient. Data were collected through clinical examinations and interviews, after examiner calibration. Overjet (OJ) was considered a risk factor when it presented values higher than 3 mm, while lip coverage was classified as adequate or inadequate. Yates' chi-squared test verified the association between the variables and odds ratio. Significance level was set at 5%. The prevalence of dental injuries was 21%. Boys experienced more injuries than girls, 21.9% and 20%, respectively (P > 0.05). Falls and collisions were the main causes of dental trauma, 63.8% and 24.5%, respectively. There was a statistically significant difference between traumatic dental injuries (TDI) and OJ (95% CI 0.22-0.63) (P < 0.001) and between TDI and inadequate lip coverage (95% CI 9.16-34.93) (P < 0.001). Data indicated that boys presenting an OJ size >3 mm and inadequate lip coverage were more likely to have TDI in Campina Grande, Brazil. PMID- 19290901 TI - Parent and caretaker knowledge about avulsion of permanent teeth. AB - Traumatic dentoalveolar injuries are frequent in children and adolescents, affecting teeth, their supporting structures and adjacent soft tissues and contributing to the appearance of major psychosocial and economic problems. Tooth avulsion, the most severe dentoalveolar lesion, is a dental emergency. The prognosis of avulsed teeth significantly depends on prompt and efficient action at the site of the accident, thus requiring that parents or caretakers be knowledgeable about the correct management of this situation. The objective of the present study was to assess the level of knowledge of parents or caretakers concerning the management of tooth avulsion and to investigate the association between level of knowledge and schooling, monthly family income and age. We interviewed 107 parents or caretakers using a 12-item questionnaire comprising objective questions whose answers received a score from 0 to 3. The results show that 99% of those interviewed would immediately seek professional help; however, 71% did not know what avulsion was. Only 3% would use milk as storage medium and 16% would attempt replantation of the avulsed tooth. The distribution of final means for the overall level of parent or caretaker knowledge was 44.63% for score 3, 15.88% for score 2, 17.99% for score 1 and 21.47% for score 0, showing a low level of knowledge concerning tooth avulsion. Schooling, monthly family income and age were not associated with the knowledge scores for any of the 12 questions. The level of parent and caretaker knowledge concerning the management of tooth avulsion is low, without association with age, schooling and monthly family income. PMID- 19290902 TI - Biocompatibility evaluation of alendronate paste in rat's subcutaneous tissue. AB - Alendronate is a known inhibitor of root resorption and the development of alendronate paste would enhance its utilization as intracanal medication. Therefore, this study aimed to investigate the biocompatibility of experimental alendronate paste in subcutaneous tissue of rats, for utilization in teeth susceptible to root resorption. The study was conducted on 15 male rats, weighing approximately 180-200 grams. The rats' dorsal regions were submitted to one incision on the median region and, laterally to the incision, the subcutaneous tissue was raised and gently dissected for introduction of two tubes, in each rat. The tubes were sealed at one end with gutta-percha and taken as control. The tubes were filled with experimental alendronate paste. The animals were killed at 7, 15 and 45 days after surgery and the specimens were processed in laboratory. The histological sections were stained with hematoxylin-eosin and analyzed by light microscopy. Scores were assigned to the inflammatory process and statistically compared by the Tukey test (P < 0.05). Alendronate paste promoted severe inflammation process at 7 days, with statistically significant difference compared to the control (P < 0.05%). However, at 15 days, there was a regression of inflammation and the presence of connective tissue with collagen fibers, fibroblasts and blood vessels was observed. After 45 days, it was observed the presence of well-organized connective tissue, with collagen fibers and fibroblasts, and few inflammatory cells. No statistical difference was observed between the control and experimental paste at 15 and 45 days. The experimental alendronate paste was considered biocompatible with subcutaneous tissue of rat. PMID- 19290903 TI - Effect of calcium hydroxide-based materials on periapical tissue healing and orthodontic root resorption of endodontically treated teeth in dogs. AB - This study evaluated periapical tissue healing and orthodontic root resorption of endodontically treated teeth sealed with calcium hydroxide in dogs. The sample consisted of three contralateral pairs of maxillary incisors and two contralateral pairs of mandibular incisors in each of two dogs using a split mouth design. After biomechanical preparation of the teeth in the first group (n = 10), a Ca(OH)(2) dressing was placed for 14 days before root canal filling with Ca(OH)(2)-based sealer (Sealapex) and gutta-percha points. In the second group (n = 10), root canals were obturated immediately after the mechanical preparation with gutta-percha points and zinc oxide and eugenol (ZOE)-based sealer (Endofill). After completion of endodontic treatment, the teeth were moved with an orthodontic appliance with a calibrated force of 200 g, reactivated every 21 days. After 105 days, the animals were killed and the teeth were removed upon completion of active treatment, without a period of recovery, and prepared for histomorphological analysis. All sections of each tooth were graded subjectively on a scale from one to four to obtain the average of the 16 histomorphological parameters analysed. Evaluation of the differences between the two treatment protocols was made with Mann-Whitney U-test. It was observed that the teeth treated with Ca(OH)(2)-based materials provided better outcomes (P = 5%), with complete repair of all root resorption areas, high rate of biological closure of the main canal and apical accessory canals by newly formed cementum, less intense and extensive chronic inflammatory infiltrate, and better organization of the periodontal ligament. Under the tested conditions, Ca(OH)(2)-based materials had a favourable action on periapical tissue healing and repair of orthodontic root resorption in endodontically treated dogs' teeth. PMID- 19290904 TI - Prevalence of dental trauma in deciduous teeth of Brazilian children. AB - The aim of this study was to assess the prevalence of dental trauma as compared to the prevalence of dental caries in children aged 3-59 months. A cross sectional study was carried out during the National Immunization Day for Polio in Recife, northeast Brazil. The sample comprised 3489 children under the age of 5. The clinical examination was performed by 123 previously trained dental students. The inter-examiner agreement for crown fracture was 90.32% and for tooth discoloration it was 83.87%. The prevalence of dental caries was assessed using the dmf-t index. The statistical methods included descriptive analysis and the chi-square test with a margin of error of 5%. A total of 56 142 teeth were examined and the prevalence of dental trauma was 14.9%. Dental fracture was the most prevalent injury observed in 516 teeth (0.9%), followed by discoloration in 191 (0.9%), intrusion in 12 (0.02%) and extrusion in five (0.008%). The most affected teeth were the upper central incisors. The prevalence of dental trauma increased significantly with age and family income (P < 0.0001). Dental trauma was most prevalent from 2 to 5 years of age. The prevalence of dental caries was 14.3%. This prevalence increased significantly with age (P < 0.0001) and with the reduction in family income (P < 0.0001). The results showed that the prevalence of dental caries and dental trauma was similar, and that both caused the same amount of damage to dental health for the target population. PMID- 19290905 TI - Traumatic dental injuries of permanent incisors in 11- to 13-year-old South African schoolchildren. AB - Traumatic dental injuries (TDI) are common in children. There are few data on prevalence of TDI in South African populations. We assessed the prevalence and causes of TDI to anterior teeth in 11- to 13-year-old South African schoolchildren through a cross-sectional study, using a random cluster sampling method. Oral examinations were performed by calibrated examiners following training for TDI to anterior permanent incisor teeth (eight teeth) using a modified version of Ellis's classification. Of the study population of 2610 children aged 11-13 years old from 26 primary schools, 1665 children participated. The response rate was 64%. More than two-thirds (64.4%) were 12 years old. One hundred and six children had a TDI (6.4%). After adjusting for the effect of age and socio-economic status, boys had an almost 2.5 (95% CI: 1.59, 3.69) times higher probability of having a traumatic dental injury than girls. The highest prevalence was in 12 year olds and in the high socio-economic status group. Most of the children had trauma to one tooth. Enamel fracture was the main type of TDI (69.1%). The majority of the TDIs were untreated (85.4%). Homes and schools were the most common places where TDIs occurred, while only 5.7% occurred on a street, road or pavement. Falls were the main cause of TDIs. Sport was the second most common cause and the third most common cause was collision with objects. The present study indicates that the prevalence of TDIs in schoolchildren is not as high as has been reported in other countries. The prevalence of TDIs in this population was relatively low. PMID- 19290906 TI - Analysis in vitro of direct bonding system with cyanoacrylate ester and orthodontic wires. AB - The aim of this study was to evaluate the tensile strength of orthodontic wires bonded onto the enamel with cyanoacrylate ester. To obtain the specimens, 120 human premolars (extracted for orthodontic or periodontal reasons) were included in acrylic blocks of rapid polymerization with three teeth each. Four groups were formed with ten specimens each. In the specimens, a dental splint model was made with cyanoacrylate ester and round stainless steel wire. In groups I, II and III, cyanoacrylate ester was used with round steel wires, with variation in diameter: 0.014 inches; 0.016 inches and 0.018 inches, respectively. In group IV, round steel wire 0.018 inches was used with photo polymerizing resin composite with previous acid etching. The adhesive force of the materials was measured in two points under the action of the tensiometer (ETM-USA). The number of loose wires was counted along with those that remained fixed according to the different levels of force applied because of the direction of the tensile force (vertical or horizontal) and the diameter of the wire used. The data obtained were first submitted to a descriptive analysis and then submitted to a statistical analysis (Friedman's Test and Dunn's Test of Multiple Comparison - Epi-info 3.2). Within the limitations of the experimental conditions presented, the cyanoacrylate ester or 'Super Bonder' maintained bonded to enamel and steel wires (0.016 and 0.018 inches) during the tensile strength tests under different levels of applied forces. PMID- 19290907 TI - New method of temporary rehabilitation after traumatic tooth loss in a juvenile patient: a case report. AB - We report a 12-year-old female patient who had experienced traumatic loss of one of her permanent central incisors. In the course of interdisciplinary treatment, the gap in the front teeth was filled with a prosthetic plastic tooth fastened to an orthodontic palatal implant by means of a wire extension (stainless steel 1.2 mm; laser-welded). A palatal implant is a temporary orthodontic anchorage device which, in contrast to a conventional single tooth implant, is not inserted in the toothless alveolar bone but in the palatal region to permit unhindered maxillary growth. The advantages of this treatment compared with traditional treatment options are that it permits secure fixation, adjustment to vertical growth and good hygiene. Besides, it is an economical treatment modality, can be used as anchorage in conjunction with orthodontic treatment and provides good comfort, esthetics and phonetics. PMID- 19290908 TI - Mandibular fracture with a mouth formed mouthguard in kickboxing. AB - Reports of injuries caused by kickboxing, one of the contact sports that potentially causes a large number of injuries, are relatively rare. Wearing a mouthguard is obligatory in kickboxing, but the association between maxillofacial injuries and the quality of mouthguards has not been described thus far. In this article, we present a case of mandibular fracture in a 25-year-old male, who was injured during kickboxing despite wearing a mouth formed mouthguard. PMID- 19290910 TI - Missile trauma in dogs. PMID- 19290911 TI - Recent topics related to human herpesvirus 6 cell tropism. AB - Human herpesvirus 6 (HHV-6) is a T lymphotropic herpes virus that is categorized into two variants, A (HHV-6A) and B (HHV-6B), on the basis of distinct genetic, immunological and biological characteristics. HHV-6 uses human CD46 as a cellular receptor. Without viral replication, HHV-6A induces cell-cell fusion between cells expressing human CD46. Some HHV-6B strains can also induce CD46-mediated cell-cell fusion. A multiple glycoprotein complex composed of glycoprotein (g) H gL complexed with gQ1 and gQ2 has been identified, and found to be a viral ligand for the human CD46 receptor. Moreover, a novel complex consisting of gH/gL/gO, which does not associate with CD46, has also been identified. The evidence suggests that an additional receptor for HHV-6B or both variants may play a role in determining the cell tropism of this virus. Finally, cholesterol in the HHV-6 envelope and plasma membrane of the host cells plays an important role in HHV-6 entry, although how this function relates to cell-envelope fusion remains to be elucidated. PMID- 19290912 TI - Borna disease virus: a unique pathogen and its interaction with intracellular signalling pathways. AB - Borna disease virus (BDV) is a neurotropic RNA virus that establishes non cytolytic persistent infection in the central nervous system of warm-blooded animals. Depending on the host species and the route of infection, BDV persistence can modulate neuronal plasticity and animal behaviour and/or may provoke a T cell-mediated immunopathological reaction with high mortality. Therefore, BDV functions as a model pathogen to study persistent virus infection in the central nervous system. Here, we review recent evidence showing that BDV interferes with a spectrum of intracellular signalling pathways, which may be involved in viral spread, maintenance of persistence and modulation of neurotransmitter pathways. PMID- 19290913 TI - A new player in a deadly game: influenza viruses and the PI3K/Akt signalling pathway. AB - Upon influenza A virus infection of cells, a wide variety of antiviral and virus supportive signalling pathways are induced. Phosphatidylinositol-3-kinase (PI3K) is a recent addition to the growing list of signalling mediators that are activated by these viruses. Several studies have addressed the role of PI3K and the downstream effector protein kinase Akt in influenza A virus-infected cells. PI3K/Akt signalling is activated by diverse mechanisms in a biphasic manner and is required for multiple functions during infection. While the kinase supports activation of the interferon regulatory factor-3 during antiviral interferon induction, it also exhibits virus supportive functions. In fact, PI3K not only regulates a very early step during viral entry but also results in suppression of premature apoptosis at later stages of infection. The latter function is dependent on the expression of the viral non-structural protein-1 (A/NS1). It has been shown that PI3K activation occurs by direct interaction of A/NS1 with the p85 regulatory subunit and interaction sites of A/NS1 and p85 have now been mapped in detail. Here, we summarize the current knowledge on influenza virus induced PI3K signalling and how this pathway supports viral propagation. PMID- 19290914 TI - Resistance of Neisseria gonorrhoeae to non-oxidative killing by adherent human polymorphonuclear leucocytes. AB - Symptomatic infection with Neisseria gonorrhoeae (Gc) is characterized by abundant neutrophil (PMN, polymorphonuclear leucocyte) influx, but PMNs cannot clear initial infection, indicating that Gc possess defences against PMN challenge. In this study, survival of liquid-grown Gc was monitored after synchronous infection of adherent, interleukin 8-treated human PMNs. 40-70% of FA1090 Gc survived 1 h of PMN exposure, after which bacterial numbers increased. Assays with bacterial viability dyes along with soybean lectin to detect extracellular Gc revealed that a subset of both intracellular and extracellular PMN-associated Gc were viable. Gc survival was unaffected in PMNs chemically or genetically deficient for producing reactive oxygen species (ROS). This result held true even for OpaB+ Gc, which stimulate neutrophil ROS production. Catalase- and RecA-deficient Gc, which are more sensitive to ROS in vitro, had no PMN survival defect. recN and ngo1686 mutant Gc also exhibit increased sensitivity to ROS and PMNs, but survival of these mutants was not rescued in ROS-deficient cells. The ngo1686 mutant showed increased sensitivity to extracellular but not intracellular PMN killing. We conclude that Gc are remarkably resistant to PMN killing, killing occurs independently of neutrophil ROS production and Ngo1686 and RecN defend Gc from non-oxidative PMN antimicrobial factors. PMID- 19290915 TI - Centrosome abnormalities during a Chlamydia trachomatis infection are caused by dysregulation of the normal duplication pathway. AB - The presence of supernumerary centrosomes in cells infected with Chlamydia trachomatis may provide a mechanism to explain the association of C. trachomatis genital infection with cervical cancer. We show that the amplified centrosomal foci induced during a chlamydial infection contain both centriolar and pericentriolar matrix markers, demonstrating that they are bona fide centrosomes. As there were multiple immature centrioles but approximately one mature centriole per cell, aborted cytokinesis alone cannot account for centrosome amplification during a chlamydial infection. Production of supernumerary centrosomes required the kinase activities of Cdk2 and Plk4, which are known regulators of centrosome duplication, and progression through S-phase, which is the stage in the cell cycle when duplication of the centrosome occurs. These requirements indicate that centrosome amplification during a chlamydial infection depends on the host centrosome duplication pathway, which normally produces a single procentriole from each template centriole. However, C. trachomatis induces a loss of numerical control so that multiple procentrioles are formed per template. PMID- 19290916 TI - HadA is an atypical new multifunctional trimeric coiled-coil adhesin of Haemophilus influenzae biogroup aegyptius, which promotes entry into host cells. AB - The Oca (Oligomeric coiled-coil adhesin) family is a subgroup of the bacterial trimeric autotransporter adhesins, which includes structurally related proteins, such as YadA of Yersinia enterocolitica and NadA of Neisseria meningitidis. In this study, we searched in silico for novel members of this family in bacterial genomes and identified HadA (Haemophilus adhesin A), a trimeric autotransporter expressed only by Haemophilus influenzae biogroup aegyptius causing Brazilian purpuric fever (BPF), a fulminant septicemic disease of children. By comparative genomics and sequence analysis we predicted that the hadA gene is harboured on a mobile genetic element unique to BPF isolates. Biological analysis of HadA in the native background was limited because this organism is not amenable to genetic manipulation. Alternatively, we demonstrated that expression of HadA confers to a non-invasive Escherichia coli strain the ability to adhere to human cells and to extracellular matrix proteins and to induce in vitro bacterial aggregation and microcolony formation. Intriguingly, HadA is predicted to lack the typical N terminal head domain of Oca proteins generally associated with cellular receptor binding. We propose here a structural model of the HadA coiled-coil stalk and show that the N-terminal region is still responsible of the binding activity and a KGD motif plays a role. Interestingly, HadA promotes bacterial entry into mammalian cells. Our results show a cytoskeleton re-arrangement and an involvement of clathrin in the HadA-mediated internalization. These data give new insights on the structure-function relationship of oligomeric coiled-coil adhesins and suggest a potential role of this protein in the pathogenesis of BPF. PMID- 19290917 TI - Kinases and phosphatases of the immune system. PMID- 19290918 TI - T-cell receptor proximal signaling via the Src-family kinases, Lck and Fyn, influences T-cell activation, differentiation, and tolerance. AB - T-cell development in the thymus and activation of mature T cells in secondary lymphoid organs requires the ability of cells to respond appropriately to environmental signals at multiple stages of their development. The process of thymocyte selection insures a functional T-cell repertoire, while activation of naive peripheral T cells induces proliferation, gain of effector function, and, ultimately, long-lived T-cell memory. The T-cell immune response is initiated upon engagement of the T-cell receptor (TCR) and coreceptor, CD4 or CD8, by cognate antigen/major histocompatibility complexes presented by antigen presenting cells. TCR/coreceptor engagement induces the activation of biochemical signaling pathways that, in combination with signals from costimulator molecules and cytokine receptors, direct the outcome of the response. Activation of the src family kinases p56(lck) (Lck) and p59(fyn) (Fyn) is central to the initiation of TCR signaling pathways. This review focuses on our current understanding of the mechanisms by which these two proteins orchestrate T-cell function. PMID- 19290920 TI - The structure, regulation, and function of ZAP-70. AB - The tyrosine ZAP-70 (zeta-associated protein of 70 kDa) kinase plays a critical role in activating many downstream signal transduction pathways in T cells following T-cell receptor (TCR) engagement. The importance of ZAP-70 is evidenced by the severe combined immunodeficiency that occurs in ZAP-70-deficient mice and humans. In this review, we describe recent analyses of the ZAP-70 crystal structure, revealing a complex regulatory mechanism of ZAP-70 activity, the differential requirements for ZAP-70 and spleen tyrosine kinase (SyK) in early T cell development, as well as the role of ZAP-70 in chronic lymphocytic leukemia and autoimmunity. Thus, the critical importance of ZAP-70 in TCR signaling and its predominantly T-cell-restricted expression pattern make ZAP-70 an attractive drug target for the inhibition of pathological T-cell responses in disease. PMID- 19290919 TI - Multiple roles of Lyn kinase in myeloid cell signaling and function. AB - Lyn is an Src family kinase present in B lymphocytes and myeloid cells. In these cell types, Lyn establishes signaling thresholds by acting as both a positive and a negative modulator of a variety of signaling responses and effector functions. Lyn deficiency in mice results in the development of myeloproliferation and autoimmunity. The latter has been attributed to the hyper-reactivity of Lyn deficient B cells due to the unique role of Lyn in downmodulating B-cell receptor activation, mainly through phosphorylation of inhibitory molecules and receptors. Myeloproliferation results, on the other hand, from the enhanced sensitivity of Lyn-deficient progenitors to a number of colony-stimulating factors (CSFs). The hyper-sensitivity to myeloid growth factors may also be secondary to poor inhibitory receptor phosphorylation, leading to impaired recruitment/activation of tyrosine phosphatases and reduced downmodulation of CSF signaling responses. Despite these observations, the overall role of Lyn in the modulation of myeloid cell effector functions is much less well understood, as often both positive and negative roles of this kinase have been reported. In this review, we discuss the current knowledge of the duplicitous nature of Lyn in the modulation of myeloid cell signaling and function. PMID- 19290921 TI - Bruton's tyrosine kinase (Btk): function, regulation, and transformation with special emphasis on the PH domain. AB - Bruton's agammaglobulinemia tyrosine kinase (Btk) is a cytoplasmic tyrosine kinase important in B-lymphocyte development, differentiation, and signaling. Btk is a member of the Tec family of kinases. Mutations in the Btk gene lead to X linked agammaglobulinemia (XLA) in humans and X-linked immunodeficiency (Xid) in mice. Activation of Btk triggers a cascade of signaling events that culminates in the generation of calcium mobilization and fluxes, cytoskeletal rearrangements, and transcriptional regulation involving nuclear factor-kappaB (NF-kappaB) and nuclear factor of activated T cells (NFAT). In B cells, NF-kappaB was shown to bind to the Btk promoter and induce transcription, whereas the B-cell receptor dependent NF-kappaB signaling pathway requires functional Btk. Moreover, Btk activation is tightly regulated by a plethora of other signaling proteins including protein kinase C (PKC), Sab/SH3BP5, and caveolin-1. For example, the prolyl isomerase Pin1 negatively regulates Btk by decreasing tyrosine phosphorylation and steady state levels of Btk. It is intriguing that PKC and Pin1, both of which are negative regulators, bind to the pleckstrin homology domain of Btk. To this end, we describe here novel mutations in the pleckstrin homology domain investigated for their transforming capacity. In particular, we show that the mutant D43R behaves similar to E41K, already known to possess such activity. PMID- 19290922 TI - Conformational snapshots of Tec kinases during signaling. AB - The control of cellular signaling cascades is of utmost importance in regulating the immune response. Exquisitely precise protein-protein interactions and chemical modification of substrates by enzymatic catalysis are the fundamental components of the signals that alert immune cells to the presence of a foreign antigen. In particular, the phosphorylation events induced by protein kinase activity must be spatially and temporally regulated by specific interactions to maintain a normal and effective immune response. High resolution structures of many protein kinases along with supporting biochemical data are providing significant insight into the intricate regulatory mechanisms responsible for controlling cellular signaling. The Tec family kinases are immunologically important kinases for which regulatory details are beginning to emerge. This review focuses on bringing together structural insights gained over the years to develop an understanding of how domain interactions both within the Tec kinases and between the Tec kinases and other signaling molecules control immune cell function. PMID- 19290923 TI - Tec kinases regulate T-lymphocyte development and function: new insights into the roles of Itk and Rlk/Txk. AB - The Tec (tyrosine kinase expressed in hepatocellular carcinoma) family of non receptor tyrosine kinases consists of five members: Tec, Bruton's tyrosine kinase (Btk), inducible T-cell kinase (Itk), resting lymphocyte kinase (Rlk/Txk), and bone marrow-expressed kinase (Bmx/Etk). Although their functions are probably best understood in antigen receptor signaling, where they participate in the phosphorylation and regulation of phospholipase C-gamma (PLC-gamma), it is now appreciated that these kinases contribute to signaling from many receptors and that they participate in multiple downstream pathways, including regulation of the actin cytoskeleton. In T cells, three Tec kinases are expressed, Itk, Rlk/Txk, and Tec. Itk is expressed at highest amounts and plays the major role in regulating signaling from the T-cell receptor. Recent studies provide evidence that these kinases contribute to multiple aspects of T-cell biology and have unique roles in T-cell development that have revealed new insight into the regulation of conventional and innate T-cell development. We review new findings on the Tec kinases with a focus on their roles in T-cell development and mature T cell differentiation. PMID- 19290924 TI - The Tec kinases Itk and Rlk regulate conventional versus innate T-cell development. AB - Tec family kinases are important components of antigen receptor signaling pathways in B cells, T cells, and mast cells. In T cells, three members of this family, inducible T-cell kinase (Itk), resting lymphocyte kinase (Rlk), and Tec, are expressed. In the absence of Itk and Rlk, T-cell receptor signaling is impaired, with defects in mitogen-activated protein kinase activation, Ca(2+) mobilization, and actin polymerization. During T-cell development in the thymus, no role has been found for these kinases in the CD4(+) versus CD8(+) T-cell lineage decision; however, several studies indicate that Itk and Rlk contribute to the signaling leading to positive and negative selection. In addition, we and others have recently described an important role for Itk and Rlk in the development of conventional as opposed to innate CD4(+) and CD8(+) T cells. Natural killer T and gammadelta T-cell populations are also altered in Itk- and Rlk/Itk-deficient mice. These findings strongly suggest that the strength of T cell receptor signaling during development determines whether T cells mature into conventional versus innate lymphocyte lineages. This lineage decision is also influenced by signaling via signaling lymphocytic activation molecule (SLAM) family receptors. Here we discuss these two signaling pathways that each contribute to conventional versus innate T-cell lineage commitment. PMID- 19290925 TI - Tyrosine kinases and their substrates in B lymphocytes. AB - Gene-targeting experiments have highlighted the importance of the intracellular protein tyrosine kinases, Lyn, Syk, and Btk, in B-cell receptor-mediated phospholipase C gamma 2 and phosphoinositide 3-kinase activation. In linking such tyrosine kinases with effector enzymes, an important role has emerged for adapter molecules. Adapter proteins nucleate formation of distinct signaling complexes in a specific location within the cell and facilitate the interaction between these signaling components in this particular subcellular compartment, which, in turn, contribute to the qualitative and quantitative control of B-cell signaling. PMID- 19290926 TI - The tyrosine kinase network regulating mast cell activation. AB - Mast cell mediator release represents a pivotal event in the initiation of inflammatory reactions associated with allergic disorders. These responses follow antigen-mediated aggregation of immunoglobulin E (IgE)-occupied high-affinity receptors for IgE (Fc epsilon RI) on the mast cell surface, a response which can be further enhanced following stem cell factor-induced ligation of the mast cell growth factor receptor KIT (CD117). Activation of tyrosine kinases is central to the ability of both Fc epsilon RI and KIT to transmit downstream signaling events required for the regulation of mast cell activation. Whereas KIT possesses inherent tyrosine kinase activity, Fc epsilon RI requires the recruitment of Src family tyrosine kinases and Syk to control the early receptor-proximal signaling events. The signaling pathways propagated by these tyrosine kinases can be further upregulated by the Tec kinase Bruton's tyrosine kinase and downregulated by the actions of the tyrosine Src homology 2 domain-containing phosphatase 1 (SHP-1) and SHP-2. In this review, we discuss the regulation and role of specific members of this tyrosine kinase network in KIT and Fc epsilon RI-mediated mast cell activation. PMID- 19290927 TI - Abl tyrosine kinases in T-cell signaling. AB - Stimulation of the T-cell antigen receptor (TCR) leads to the activation of signaling pathways that are essential for T-cell development and the response of mature T cells to antigens. The TCR has no intrinsic catalytic activity, but TCR engagement results in tyrosine phosphorylation of downstream targets by non receptor tyrosine kinases. Three families of tyrosine kinases have long been recognized to play critical roles in TCR-dependent signaling. They are the Src, zeta-associated protein of 70 kDa, and Tec families of kinases. More recently, the Abelson (Abl) tyrosine kinases have been shown to be activated by TCR engagement and to be required for maximal TCR signaling. Using T-cell conditional knockout mice deficient for Abl family kinases, Abl (Abl1) and Abl-related gene (Arg) (Abl2), it was recently shown that loss of Abl kinases results in defective T-cell development and a partial block in the transition to the CD4(+)CD8(+) stage. Abl/Arg double null T cells exhibit impaired TCR-induced signaling, proliferation, and cytokine production. Moreover, conditional knockout mice lacking Abl and Arg in T cells exhibit impaired CD8(+) T-cell expansion in vivo upon Listeria monocytogenes infection. Thus, Abl kinase signaling is required for both T-cell development and mature T-cell function. PMID- 19290928 TI - Calmodulin-dependent phosphatase, kinases, and transcriptional corepressors involved in T-cell activation. AB - The second messenger calcium plays an essential role in mediating the T-cell receptor (TCR) signaling pathway leading to cytokine production and T-cell clonal expansion. The immunosuppressive drugs cyclosporine A and FK506 have served both as therapeutic agents and as molecular probes for unraveling the protein phosphatase calcineurin as a rate-limiting enzyme involved in the transmission of calcium signal from the cytosol into the nucleus to reprogram gene expression. The use of mouse knockout models has helped to verify and further elucidate the functions of different isoforms of calcineurin in both helper T-cell activation and thymocyte development. In addition to calcineurin, three other classes of calmodulin-binding proteins have also been shown to play important roles in calcium signaling in T cells. Thus, Cabin1 and class II histone deacetylases have been found to constitute a novel calcium-signaling module in conjunction with the transcription factor myocyte enhance factor family and the transcriptional coactivator p300 to suppress and activate cytokine gene transcription in a calcium-dependent manner. The calmodulin-dependent protein kinases II and IV were also shown to play negative and positive regulatory functions, respectively, in TCR-mediated cytokine production. The crosstalks among these and other signal transducers in T cells form an extensive nonlinear signaling network that dictates the final outcome of the TCR signaling pathway. PMID- 19290929 TI - CARMA1-mediated NF-kappaB and JNK activation in lymphocytes. AB - Activation of transcription factor nuclear factor-kappaB (NF-kappaB) and Jun N terminal kinase (JNK) play the pivotal roles in regulation of lymphocyte activation and proliferation. Deregulation of these signaling pathways leads to inappropriate immune response and contributes to the development of leukemia/lymphoma. The scaffold protein CARMA1 [caspase-recruitment domain (CARD) membrane-associated guanylate kinase (MAGUK) protein 1] has a central role in regulation of NF-kappaB and the JNK2/c-Jun complex in both B and T lymphocytes. During last several years, tremendous work has been done to reveal the mechanism by which CARMA1 and its signaling partners, B cell CLL-lymphoma 10 and mucosa associated lymphoid tissue 1, are activated and mediate NF-kappaB and JNK activation. In this review, we summarize our findings in revealing the roles of CARMA1 in the NF-kappaB and JNK signaling pathways in the context of recent advances in this field. PMID- 19290930 TI - Regulation of the immune response by stress-activated protein kinases. AB - Activation of immune cells to mediate an immune response is often triggered by potential 'danger' or 'stress' stimuli that the organism receives. Within the mitogen-activated protein kinases (MAPKs) family, the stress-activated protein kinase (SAPK) group was defined as group of kinases that activated by stimuli that cause cell stress. In the immune cells, SAPKs are activated by antigen receptors (B- or T-cell receptors), Toll-like receptors, cytokine receptors, and physical-chemical changes in the environment among other stimuli. The SAPKs are established to be important mediators of intracellular signaling during adaptive and innate immune responses. Here we summarize what is currently known about the role of two sub-groups of SAPKs - c-Jun NH(2)-terminal kinase and p38 MAPK-in the function of specific components of the immune system and the overall contribution to the immune response. PMID- 19290931 TI - TNFR signaling: ubiquitin-conjugated TRAFfic signals control stop-and-go for MAPK signaling complexes. AB - Nearly two decades after the initial cloning and identification of the founding father of the tumor necrosis factor receptor (TNFR) family, much has been learned about the mechanisms by which these receptors signal to critical transcription factors and other targets that regulate gene expression and cellular physiology. Mitogen-activated protein kinases (MAPKs) and inhibitor of nuclear factor (NF) kappaB (I kappaB) kinases (IKKs) were identified early on as the upstream kinases responsible for activation of activator-protein 1 (AP-1) and NF-kappaB, respectively, and later on for their ability to control life-or-death decisions in TNF-stimulated cells. Both of these critical pathways are regulated at the level of MAPK kinase kinases (MAP3Ks), after which point they diverge. Recent work, however, illustrates that protein ubiquitination cascades play a critical initiating role in TNFR signaling and account for spatial and temporal separation of IKK and MAPK signaling cascades and thereby determine biological specificity and outcome. Cellular inhibitors of apoptosis (cIAPs) 1 and 2 are ubiquitin (Ub) ligases (E3s) that mediate canonical Lys48-linked ubiquitination of TNFR associated factor 3 (TRAF3), marking it for subsequent degradation by the proteasome. TRAF3 degradation releases the brake on TRAF2/6:MAP3K signaling complexes responsible for MAPK activation, leading to their translocation from the cytoplasmic segment of the receptor to the cytosol where they initiate MAPK phosphorylation and activation. By contrast, IKK activation proceeds considerably faster than MAPK activation, takes place at the receptor, and is independent of cIAP1/2 activity and TRAF3 degradation. This arrangement may be important for ensuring the proper delivery of NF-kappaB-dependent survival signals and conversion of JNK-promoted death signals to proliferative ones. PMID- 19290932 TI - New insights into the regulation and function of serine/threonine kinases in T lymphocytes. AB - The development of T lymphocytes in the thymus and the function of mature T cells in adaptive immune responses are choreographed by antigen receptors, costimulatory molecules, adhesion molecules, cytokines, and chemokines. These extrinsic stimuli are coupled to a diverse network of signal transduction pathways that control the transcriptional and metabolic programs that determine T cell function. At the core of T-lymphocyte signal transduction is the regulated metabolism of inositol phospholipids and the production of two key lipid second messengers: polyunsaturated diacylglycerols (DAGs) and phosphatidylinositol (3-5) triphosphate [PI-(3-5)-P(3)]. The object of the present review is to discuss facts, controversies, and unresolved issues about DAG and PI-(3,4,5)-P(3) production in T lymphocytes and to discuss some of the serine/threonine kinases that control unique aspects of T-lymphocyte biology and coordinate T-cell participation in adaptive immune responses. PMID- 19290933 TI - Fine tuning the immune response with PI3K. AB - The phosphoinositide 3-kinase (PI3K) family of lipid kinases regulates diverse aspects of lymphocyte behavior. This review discusses how genetic and pharmacological tools have yielded an increasingly detailed understanding of how PI3K enzymes function at different stages of lymphocyte development and activation. Following antigen receptor engagement, activated PI3K generates 3 phosphorylated inositol lipid products that serve as membrane targeting signals for numerous proteins involved in the assembly of multiprotein complexes, termed signalosomes, and immune synapse formation. In B cells, class IA PI3K is the dominant subgroup whose loss causes profound defects in development and antigen responsiveness. In T cells, both class IA and IB PI3K contribute to development and immune function. PI3K also regulates both chemokine responsiveness and antigen-driven changes in lymphocyte trafficking. PI3K modulates the function not only of effector T cells, but also regulatory T cells; these disparate functions culminate in unexpected autoimmune phenotypes in mice with PI3K-deficient T cells. Thus, PI3K signaling is not a simple switch to promote cellular activation, but rather an intricate web of interactions that must be properly balanced to ensure appropriate cellular responses and maintain immune homeostasis. Defining these complexities remains a challenge for pharmaceutical development of PI3K inhibitors to combat inflammation and autoimmunity. PMID- 19290934 TI - Janus kinases in immune cell signaling. AB - The Janus family kinases (Jaks), Jak1, Jak2, Jak3, and Tyk2, form one subgroup of the non-receptor protein tyrosine kinases. They are involved in cell growth, survival, development, and differentiation of a variety of cells but are critically important for immune cells and hematopoietic cells. Data from experimental mice and clinical observations have unraveled multiple signaling events mediated by Jaks in innate and adaptive immunity. Deficiency of Jak3 or Tyk2 results in defined clinical disorders, which are also evident in mouse models. A striking phenotype associated with inactivating Jak3 mutations is severe combined immunodeficiency syndrome, whereas mutation of Tyk2 results in another primary immunodeficiency termed autosomal recessive hyperimmunoglobulin E syndrome. By contrast, complete deletion of Jak1 or Jak2 in the mouse are not compatible with life and, unsurprisingly, do not have counterparts in human disease. However, activating mutations of each of the Jaks are found in association with malignant transformation, the most common being gain-of-function mutations of Jak2 in polycythemia vera and other myeloproliferative disorders. Our existing knowledge on Jak signaling pathways and fundamental work on their biochemical structure and intracellular interactions allow us to develop new strategies for controlling autoimmune diseases or malignancies by developing selective Jak inhibitors, which are now coming into clinical use. Despite the fact that Jaks were discovered only a little more than a decade ago, at the time of writing there are 20 clinical trials underway testing the safety and efficacy of Jak inhibitors. PMID- 19290936 TI - PEST family phosphatases in immunity, autoimmunity, and autoinflammatory disorders. AB - The proline-, glutamic acid-, serine- and threonine-rich (PEST) family of protein tyrosine phosphatases (PTPs) includes proline-enriched phosphatase (PEP)/lymphoid tyrosine phosphatase (LYP), PTP-PEST, and PTP-hematopoietic stem cell fraction (HSCF). PEP/LYP is a potent inhibitor of T-cell activation, principally by suppressing the activity of Src family protein tyrosine kinases (PTKs). This function seems to be dependent, at least in part, on the ability of PEP to bind C terminal Src kinase (Csk), a PTK also involved in inactivating Src kinases. Interestingly, a polymorphism of LYP in humans (R620W) is a significant risk factor for autoimmune diseases including type 1 diabetes, rheumatoid arthritis, and lupus. The R620W mutation may be a 'gain-of-function' mutation. In non hematopoietic cells, PTP-PEST is a critical regulator of adhesion and migration. This effect correlates with the aptitude of PTP-PEST to dephosphorylate cytoskeletal proteins such as Cas, focal adhesion associated-kinase (FAK), Pyk2, and PSTPIP. While not established, a similar function may also exist in immune cells. Additionally, overexpression studies provided an indication that PTP-PEST may be a negative regulator of lymphocyte activation. Interestingly, mutations in a PTP-PEST- and PTP-HSCF-interacting protein, PSTPIP1, were identified in humans with pyogenic sterile arthritis, pyoderma gangrenosum, and acne (PAPA) syndrome and familial recurrent arthritis, two autoinflammatory diseases. These mutations abrogate the ability of PSTPIP1 to bind PTP-PEST and PTP-HSCF, suggesting that these two PTPs may be negative regulators of inflammation. PMID- 19290935 TI - CD45, CD148, and Lyp/Pep: critical phosphatases regulating Src family kinase signaling networks in immune cells. AB - Reciprocal regulation of tyrosine phosphorylation by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs) is central to normal immune cell function. Disruption of the equilibrium between PTK and PTP activity can result in immunodeficiency, autoimmunity, or malignancy. Src family kinases (SFKs) play a central role in both immune cell function and disease due to their proximal position in numerous signal transduction cascades including those emanating from integrin, T and B-cell antigen receptors, Fc, growth factor, and cytokine receptors. Given that tight regulation of SFKs activity is critical for appropriate responses to stimulation of these various signaling pathways, it is perhaps not surprising that multiple PTPs are involved in their regulation. Here, we focus on the role of three phosphatases, CD45, CD148, and LYP/PEP, which are critical regulators of SFKs in hematopoietic cells. We review our current understanding of their structures, expression, functions in different hematopoietic cell subsets, regulation, and putative roles in disease. Finally, we discuss remaining questions that must be addressed if we are to have a clearer understanding of the coordinated regulation of tyrosine phosphorylation and signaling networks in hematopoietic cells and how they could potentially be manipulated therapeutically in disease. PMID- 19290937 TI - T-cell protein tyrosine phosphatase is a key regulator in immune cell signaling: lessons from the knockout mouse model and implications in human disease. AB - The immune system requires for its proper ontogeny, differentiation, and maintenance the function of several tyrosine kinases and adapters that create and modify tyrosine phosphorylation sites. Tyrosine phosphorylation is a crucial protein modification in immune cell signaling and can be reversed by protein tyrosine phosphatases (PTPs). Much progress has been made in identifying and understanding PTP function in the immune system. In this review, we present one of these proteins, named T-cell PTPs (TC-PTP) (gene name PTPN2), a classical, non receptor PTP that is ubiquitously expressed with particularly high expression in hematopoietic tissues. TC-PTP is remarkable not only by the fact that it appears to influence most, if not all, cells involved in the development of the immune system, from stem cells to differentiated lineages, but also recent findings have positioned it at the core of several human diseases from autoimmune disease to cancer. PMID- 19290939 TI - Functional characterization of the muscarinic receptors involved in endothelium dependent relaxation in isolated canine uterine artery. AB - Acetylcholine interacts with endothelial muscarinic receptors releasing nitric oxide and causing vasodilatation. To identify the receptor subtype responsible for acetylcholine-induced relaxation in canine uterine artery, the usual organ bath method for in vitro investigation on isolated blood vessels was applied. Using a range of muscarinic receptor antagonists such as atropine (nonselective), pirenzepine (M(1)-selective), methoctramine (M(2)-selective) and p-fluoro hexahydro-sila-difenidol (p-FHHSiD) (M(1)/M(3)) and determining pA2 value of those antagonists through Shild analysis, we aimed at establishing a precise receptor mechanism underlying acetylcholine-induced relaxation in isolated canine uterine artery. The relaxation of uterine arterial rings in response to acetylcholine in the presence or absence of selective muscarinic receptors antagonists was calculated using concentration response curves. Acetylcholine induced concentration-dependent and endothelium-dependent relaxation of arterial rings precontracted with phenylephrine (pEC(50) = 6.90 +/- 0.02). Muscarinic receptors antagonists atropine, pirenzepine, methoctramine and p-FHHSiD competitively antagonized the response to acetylcholine and obtained pA(2) values were 9.91 +/- 0.06, 6.60 +/- 0.04, 6.21 +/- 0.08 and 8.05 +/- 0.1, respectively. This study showed that acetylcholine induced endothelium-dependent relaxation of canine uterine artery by stimulation of muscarinic receptors localized on the endothelial cells. On the basis of differential antagonist affinity, we suggest that the muscarinic receptors involved in the acetylcholine-induced relaxation of canine uterine artery are predominantly of M(3) subtype. PMID- 19290940 TI - Hydrogenated castor oil nanoparticles as carriers for the subcutaneous administration of tilmicosin: in vitro and in vivo studies. AB - Tilmicosin-loaded solid lipid nanoparticles (SLN) were prepared with hydrogenated castor oil (HCO) by o/w emulsion-solvent evaporation technique. The nanoparticle diameters, surface charges, drug loadings and encapsulation efficiencies of different formulations were 90 approximately 230 nm, -6.5 approximately -12.5 mV, 40.3 approximately 59.2% and 5.7 approximately 11.7% (w/w), respectively. In vitro release studies of the tilmicosin-loaded nanoparticles showed a sustained release and the released tilmicosin had the same antibacterial activity as that of the free drug. Pharmacokinetics study after subcutaneous administration to Balb/c mice demonstrated that a single dose of tilmicosin-loaded nanoparticles resulted in sustained serum drug levels (>0.1 microg/mL) for 8 days, as compared with only 5 h for the same amount of tilmicosin phosphate solution. The time to maximum concentration (Tmax), half-life of absorption (T(1/2) ab) and half-life of elimination (T(1/2) el) of tilmicosin-loaded nanoparticles were much longer than those of tilmicosin phosphate solution. Tissue section showed that drug loaded nanoparticles caused no inflammation at the injection site. Cytotoxicity study in cell culture and acute toxicity test in mice demonstrated that the nanoparticles had little or no toxicity. The results of this exploratory study suggest that the HCO-SLN could be a useful system for the delivery of tilmicosin by subcutaneous administration. PMID- 19290938 TI - SHP-1 and SHP-2 in T cells: two phosphatases functioning at many levels. AB - Tyrosine phosphorylation and dephosphorylation of proteins play a critical role for many T-cell functions. The opposing actions of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs) determine the level of tyrosine phosphorylation at any time. It is well accepted that PTKs are essential during T cell signaling; however, the role and importance of PTPs are much less known and appreciated. Both transmembrane and cytoplasmic tyrosine phosphatases have been identified in T cells and shown to regulate T-cell responses. This review focuses on the roles of the two cytoplasmic PTPs, the Src-homology 2 domain (SH2) containing SHP-1 and SHP-2, in T-cell signaling, development, differentiation, and function. PMID- 19290941 TI - Pharmacokinetics of ketamine and its metabolite norketamine administered at a sub anesthetic dose together with xylazine to calves prior to castration. AB - The objective of this study was to evaluate the plasma pharmacokinetics of ketamine and its active metabolite norketamine administered intravenously at a dose of 0.1 mg/kg together with xylazine (0.05 mg/kg) to control the pain associated with castration in calves. A two-compartment model with an additional metabolite compartment linked to the central compartment was used to simultaneously describe the time-concentration profiles of both ketamine and its major metabolite norketamine. Parameter values estimated from the time concentration profiles observed in this study were volume of the central compartment (V(c) = 132.82 +/- 68.23 mL/kg), distribution clearance (CL(D) = 15.49 +/- 2.56 mL/min/kg), volume of the peripheral compartment (V(T) = 257.05 +/ 41.65 mL/kg), ketamine clearance by the formation of the norketamine metabolite (CL(2M) = 8.56 +/- 7.37 mL/kg/min) and ketamine clearance by other routes (CL(o) = 16.41 +/- 3.42 mL/kg/min). Previously published data from rats suggest that the metabolite norketamine contributes to the analgesic effect of ketamine, with a potency that is one-third of the parent drug. An understanding of the time concentration relationships and the disposition of the parent drug and its metabolite is therefore important for a better understanding of the analgesic potential of ketamine in cattle. PMID- 19290942 TI - Pharmacokinetics of an immediate and extended release oral morphine formulation utilizing the spheroidal oral drug absorption system in dogs. AB - This study investigated the pharmacokinetics of a human-labeled oral morphine formulation consisting of both immediate and extended release components in dogs. In a randomized design, 14 dogs were administered either 1 or 2 mg/kg morphine orally. Blood samples were collected up to 24 h post drug administration. Plasma concentrations of morphine were measured using high-pressure liquid chromatography with electrochemical coulometric detection. For both groups, maximal concentration occurred at 3 h post drug administration followed by a gradual decrease in morphine concentration over 24 h. There was substantial variability in morphine concentrations among dogs. The higher dose group produced a greater exposure (higher area-under-the-curve), higher peak concentration, longer half-life and a shorter time to peak concentration (t(max)). The specific oral morphine formulation used in this study produced sustained plasma morphine concentrations over 24 h compared with previous intravenous dosing and immediate release oral morphine studies. However, the low morphine plasma concentrations and high variability produced from this formulation, suggest that the clinical application of this formulation at the doses evaluated in this study are limited. PMID- 19290943 TI - Transsplenic portal catheterization combined with a jugular double-lumen catheter for pharmacokinetic and presystemic metabolization studies in pigs. AB - The reliability of a silicone double-lumen catheter implanted into the external jugular vein and tunnelled towards the neck region was investigated in eight pigs. Surgery was uneventful without interference with the normal homoeostasis during 8 days. After injection of amoxicillin/clavulanic acid through the distal port of the catheter, analysis of drug components in the simultaneous blood samples obtained by the proximal port and a Venoject system were comparable in one pig. Histological control of the catheterized jugular veins pointed to an acceptable tissue reaction while bacteriological examination of the tip of the catheters was negative in only three animals. A moulding of the intestinal veins was made in a pig cadaver to determine the optimal length of insertion of a silicone portal catheter from the splenic vein towards the portal vein. Surgery was straightforward in four pigs whereby the catheter was exteriorized towards the back region. No complications were encountered during and after surgery for 9 days. The technique of a double-lumen catheter placed into the jugular vein and a transsplenic portal catheter is a useful tool for the study of the pharmacokinetics and also the first-pass effect of drugs in experimental pigs. PMID- 19290944 TI - Evaluation of two scopolamine and physostigmine pretreatment regimens against nerve agent poisoning in the dog. AB - Currently, there is no viable protection against chemical warfare agents for the working dog. Physostigmine (PHY) and scopolamine (SCO) have been shown to protect dogs against nerve agents with minimal side effects. The goal of this study was to investigate whether reported protective SCO/PHY plasma concentrations of 0.2 and 0.7 ng/mL, respectively, could be reached and maintained with minimal side effects thereby identifying possible pretreatment regimens. Two continuous regimens of SCO/PHY were administered to beagle dogs. The first regimen consisted of administering transdermal SCO and intraocular PHY, the second consisted of transdermal SCO and rectal PHY. SCO/PHY plasma concentrations for each regimen were determined, individual protective times were calculated and a computerized pharmacokinetic analysis was performed. The results showed transdermal SCO and intraocular PHY routes of delivery achieved sustained protective drug concentrations with minimal side-effects and the rectal route of delivery did not. Group median protective times for the first regimen were 54.45 h for SCO and 64.35 h for PHY, and for the second regimen 63.75 h for SCO and 0 h for PHY. The combined transdermal patch and intraocular regimen may provide a safe and effective regimen against nerve agent poisoning in dogs. PMID- 19290945 TI - Pharmacokinetic behavior in sheep and cattle of 5-chloro-2-(methylthio)-6-(1 naphthyloxy)-1H-benzimidazole, a new fasciolicide agent. AB - The physicochemical properties, pK(a), Log P and solubility of compound alpha, (5 chloro-2-(methylthio)-6-(1-naphthyloxy)-1H-benzimidazole), a new fasciolicide agent, were characterized using conventional methods. Also, its pharmacokinetics was evaluated in sheep and cattle. In both species an oral dose of 12 mg/kg was administered. Blood samples were collected during 144 h and analyzed by using an HPLC assay. Results showed that compound alpha is a weak base with a pK(a) value of 2.87 and log P of 1.44. The solubility was very low in aqueous solvents. Pharmacokinetic studies showed that in both species compound alpha could not be detected at any sampling time. The mean half-life (t(1/2)) values of alpha sulphoxide in sheep and cattle were 19.86 and 29.87 h, while the half-life values of alpha sulphone were 19.43 and 46.32 h respectively. C(max) values of alpha sulphoxide did not differ between species while alpha sulphone values were higher in cattle. Plasma protein binding of alpha sulphoxide was between 82% and 86%. These results, combined with the previous efficacy studies, suggest that compound alpha could be a promising fasciolicide agent. PMID- 19290946 TI - The formation of aminorex in racehorses following levamisole administration. A quantitative and chiral analysis following synthetic aminorex or levamisole administration vs. aminorex-positive samples from the field: a preliminary report. AB - Beginning in 2004, the horseracing industry experienced an epidemic of drug positives for the amphetamine-like drug aminorex. Investigation of the therapeutic treatment of the horses called positive for this drug suggested that its source was from the administration of the anthelmintic levamisole. This study examines the urine concentrations of aminorex as a function of time following administration of synthetic, racemic aminorex. Confirmation of the presence of aminorex in urine samples from the horses known to be treated with levamisole is also presented as are data concerning the concentrations of aminorex in positives called from the field and the corresponding concentrations of levamisole found in the same samples. Furthermore, this study illustrates that the chiral isomer distribution of aminorex found in samples from the field is significantly different from that arising from the administration of synthetic, racemic aminorex and is similar to that observed from aminorex arising from levamisole administration. An examination of the chiral isomer distribution of aminorex and a determination of the presence of levamisole in a sample may be used to assess the source of an aminorex positive, distinguishing it from an intentional synthetic, racemic aminorex administration. The role of levamisole in aminorex formation is also discussed. PMID- 19290947 TI - P-glycoprotein in intestines, liver, kidney and lymphocytes in horse. AB - P-glycoprotein (P-gp) is an important drug transporter, which is expressed in a variety of cells, such as the intestinal enterocytes, the hepatocytes, the renal tubular cells and the intestinal and peripheral blood lymphocytes. We have studied the localization and the gene and protein expression of P-gp in these cells in horse. In addition we have compared the protein sequence of P-gp in horse with the protein sequences of P-gp in several other species. Real time RT PCR and Western blot showed gene and protein expression of horse P-gp in all parts of the intestines, but there was no strict correlation between these parameters. Immunohistochemistry showed localization of P-gp in the apical cell membranes of the enterocytes and, in addition, staining was observed in the intestinal intraepithelial and lamina propria lymphocytes. Peripheral blood lymphocytes also stained for P-gp, and gene and protein expression of P-gp were observed in these cells. There was a high gene and protein expression of P-gp in the liver, with P-gp-immunoreactivity in the bile canalicular membranes of the hepatocytes. Gene and protein expression of P-gp were found in the kidney with localization of the protein in different parts of the nephrons. Protein sequence alignment showed that horse P-gp has two amino acid insertions at the N-terminal region of the protein, which are not present in several other species examined. One of these is a 99 amino acid long sequence inserted at amino acid positions 23 121 from the N-terminal. The other is a six amino acid long sequence present at the amino acid positions 140-145 from the N-terminal. The results of the present study indicate that P-gp has an important function for oral bioavailability, distribution and excretion of substrate compounds in horse. PMID- 19290948 TI - Milrinone and theophylline act as lower oesophageal sphincter relaxing agents: a comparative pharmacodynamic study in the rabbit. AB - This study demonstrates that the inotropic agent milrinone and the bronchodilator drug theophylline exert a relaxing effect on the rabbit lower oesophageal sphincter in vitro. The relaxing effect of milrinone and theophylline, which is concentration-dependent, involves a second messenger 3',5'-cyclic adenosine monophosphate pathway and most probably it is accomplished through inhibition of phosphodiesterase (PDE) type III, as according to the obtained results it is not significantly modified either by nicotinic acid, an inhibitor of adenylate cyclase, or by the inhibitor of nitric oxide-synthetase N(omega)-nitro-L-arginine methylester and the purinergic antagonist suramin; moreover, it persists under non-adrenergic non-cholinergic conditions and it is both hexamethonium- and tetrodotoxin-insensitive. Both milrinone and theophylline display equal efficacy, comparable to that of the calcium blocker verapamil and the non-selective PDE inhibitor papaverine, but milrinone appears 50 times more potent than theophylline and three times less potent than verapamil, as, according to the pIC(50) values the potency rank of order is found to be verapamil (5.56) > milrinone (5.12) > theophylline (3.42). The here obtained pharmacodynamic profiles of the drugs suggest that both milrinone and theophylline may be considered as potent relaxing agents of the lower oesophageal sphincter. PMID- 19290949 TI - A step towards effect-site target-controlled infusion with propofol in dogs: a k(e0) for propofol. AB - Target-controlled infusion (TCI) anesthesia using target effect-site concentration rather than plasma concentration provides less drug consumption, safer anesthesia, less undesired side effects and improved animal welfare. The aim of this study was to calculate the constant that converts propofol plasma into effect-site concentration (k(e0)) in dogs, and to implement it in a TCI system and compare it with the effect on the central nervous system (CNS). All dogs were subjected to general anesthesia using propofol. Fourteen dogs were used as the pilot group to calculate k(e0), using the t(peak) method. Fourteen dogs were used as the test group to test and validate the model. RUGLOOP II software was used to drive the propofol syringe pump and to collect data from S/5 Datex monitor and cerebral state monitor. The calculated k(e0) was incorporated in an existing pharmacokinetic model (Beths Model). The relationship between propofol effect site concentrations and anesthetic planes, and propofol plasma and effect site concentrations was compared using Pearson's correlation analysis. Average t(peak) was 3.1 min resulting in a k(e0) of 0.7230 min(-1). The test group showed a positive correlation between anesthetic planes and propofol effect-site concentration (R = 0.69; P < 0.0001). This study proposes a k(e0) for propofol with results that demonstrated a good adequacy for the pharmacokinetic model and the measured effect. The use of this k(e0) will allow an easier propofol titration according to the anesthetic depth, which may lead to a reduction in propofol consumption and less undesired side effects usually associated to high propofol concentrations in dogs. PMID- 19290950 TI - Regulation of platelet activating factor-induced equine platelet activation by intracellular kinases. AB - Lipopolysaccharide (LPS) can activate equine platelets directly or indirectly, via leukocyte-derived platelet activating factor (PAF). Thromboxane (Tx) production by LPS-stimulated equine platelets requires p38 MAPK and this kinase has been suggested as a therapeutic target in endotoxaemia. The present study has utilised selective inhibitors to investigate the role of p38 MAPK and two other kinases, phosphatidylinositol-3 kinase (PI3K) and protein kinase C (PKC), in regulating PAF-induced Tx production, aggregation and 5-HT release in equine platelets, and the modification of these responses by LPS. LPS enhanced PAF induced 5-HT release, an effect that was reduced by the p38 MAPK inhibitor, SB203580 (60 +/- 8% reduction; n = 6). SB203580 did not affect responses to PAF alone; whereas inhibition of PKC reduced PAF-induced 5-HT release, Tx production and aggregation (maximal inhibition by the PKCdelta inhibitor, rottlerin: 69 +/- 13%, 63 +/- 14% and 97 +/- 1%, respectively; n = 6). Wortmannin and LY249002, which inhibit PI3K, also caused significant inhibition of PAF-induced aggregation (maximal inhibition 78 +/- 3% and 88 +/- 2%, respectively; n = 6). These data suggest that inhibition of platelet p38 MAPK may be of benefit in equine endotoxaemia by counteracting some of the effects of LPS. However, detrimental effects of platelet activation mediated by PAF and not enhanced by LPS are unlikely to be markedly affected. PMID- 19290951 TI - Detection and pharmacokinetics of tetrahydrogestrinone in horses. AB - The anti-doping rules of national and international sport federations ban any use of tetrahydrogestrinone (THG) in human as well as in horse sports. Initiated by the THG doping scandals in human sports a method for the detection of 3-keto 4,9,11-triene steroids in horse blood and urine was developed. The method comprises the isolation of the analytes by a combination of solid phase and liquid-liquid extraction after hydrolysis and solvolysis of the steroid conjugates. The concentrations of THG in blood and urine samples were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A THG excretion study on horses was conducted to verify the method capability for the analysis of postadministration urine samples. In addition, blood samples were collected to allow for determination of the pharmacokinetics of THG in horses. Following the administration of a single oral dose of 25 microg THG per kg bodyweight to 10 horses, samples were collected at appropriate intervals. The plasma levels of THG reached maximal concentrations of 1.5-4.8 ng/mL. Twenty-four hours after the administration plasma levels returned to baseline. In urine, THG was detectable for 36 h. Urinary peak concentrations of total THG ranged from 16 to 206 ng/mL. For the 10 horses tested, the mean plasma clearance of THG was 2250 mL/h/kg and the plasma elimination half-life was 1.9 h. PMID- 19290952 TI - Antacids in the initial management of metallic zinc ingestion in dogs. AB - Zinc poisoning in dogs, following ingestion of post-1982 U.S. one cent coins is an increasingly common toxicological syndrome causing gastrointestinal abnormalities, hemolytic anemia, pancreatitis and renal failure. Thermodynamic laws predict that the rate of the chemical reaction between HCl and metallic zinc, which releases absorbable zinc anions, is dependent on pH. The significance of the relation between pH and dissolution is, however, apparently contradicted by the fact that recent veterinary toxicology texts are silent on the use of antacids in the early management of zinc ingestion in dogs. A series of experiments were conducted to test the hypotheses that the degree to which zinc dissolution is pH dependent is likely to be of clinical importance and that the use of antacids will be of benefit in preventing zinc poisoning in dogs that had ingested metallic zinc. Zinc dissolution was strongly dependent on pH in an exponential manner, indicating that clinically achievable upward adjustment of gastric pH using antacids is likely to have significant effects on the rate of zinc absorption in dogs. These data clearly support the use of antacids during the initial treatment of metallic zinc ingestion in dogs. PMID- 19290960 TI - Hospital volume and neonatal mortality among very low birth weight infants. PMID- 19290961 TI - Neonatal outcome of VLBW infants in Berlin, Germany. PMID- 19290963 TI - 'Universal' patient consent form for case reports. PMID- 19290964 TI - 'It couldn't hurt .... Could it?' Safety of complementary and alternative medicine practices. PMID- 19290965 TI - Plasmalogen levels in full-term neonates. AB - AIM: Plasmalogens are phospholipids characterized by the presence of a vinyl ether bond at the sn-1 position of the glycerol backbone. They are particularly abundant in the nervous system, the heart and striated muscle. Peroxisomes are essential for their biosynthesis and red blood cell (RBC) plasmalogen levels are a reliable test in the investigation of patients suspect for a peroxisomal defect. The functions attributed to them include protection against oxidative stress, myelin formation and signal transduction. The aim of the present study was the investigation of RBC plasmalogen levels in neonates. METHODS: A total of 25 healthy full-term, appropriate for gestational age neonates were studied. RBC plasmalogens were estimated using gas chromatography within the first five days of life. Fifteen healthy children 1-8-year olds served as controls. RESULTS: Statistically significant lower plasmalogen levels were found in neonates compared to older children. CONCLUSION: Our results indicate that a different range of normal values for plasmalogen levels should be used in the investigation of peroxisomal diseases in neonates. The lower levels of plasmalogens in neonates found in our study could render them more vulnerable to oxidative stress. PMID- 19290969 TI - Abstracts of the 5th ELITA-ELTR (European Society for Organ Transplantation European Liver and Intestine Transplant Association) Winter Meeting, 19-21 March 2009. Are, Sweden. PMID- 19290970 TI - The 'deep dynamics' of the discipline of mental health nursing. PMID- 19290971 TI - Mental health nurses establishing psychosocial interventions within acute inpatient settings. AB - Acute inpatient units provide care for the most acutely unwell people experiencing a mental illness. As a result, the focus for care is on the containment of difficult behaviour and the management of those considered to be 'at high risk' of harm. Subsequently, recovery-based philosophies are being eroded, and psychosocial interventions are not being provided. Despite the pivotal role that mental health nurses play in the treatment process in the acute inpatient setting, a review of the literature indicates that mental health nursing practice is too custodial, and essentially operates within an observational framework without actively providing psychosocial interventions. This paper will discuss the problems with mental health nursing practice in acute inpatient units highlighted in the current literature. It will then put forward the argument for routine use of psychosocial interventions as a means of addressing some of these problems. PMID- 19290972 TI - The feeling of being trapped in and ashamed of one's own body: A qualitative study of women who suffer from eating difficulties. AB - The aim of this study was to explore women's bodily experiences of suffering from eating difficulties (ED). The research question was: How do women who suffer from ED experience the bodily aspects related to their condition? Women suffering from ED experience problems in both the physical and emotional areas. Few qualitative studies have specifically addressed sufferers' bodily experiences related to ED. An explorative design was used. The data were collected by means of focus group interviews on the subject of ED-related problems, guilt and shame, and being a mother. The interpretation of the qualitative data was inspired by hermeneutic content analysis. The main theme, 'Powerful feelings of being trapped in and ashamed of one's own body', comprised two themes: 'The feeling of being trapped by overwhelming physical sensations' and 'The feeling of being ashamed of one's own body'. Bodily experiences were reported as strong. One clinical implication for nurses is to acknowledge this fact and make it possible for these women to articulate their difficulties, especially those connected to the feelings of being trapped and ashamed. Support groups focusing on these themes could be one way of assisting women and easing some of the negative feelings associated with ED. PMID- 19290973 TI - Learning from experience: Using action research to discover consumer needs in post-seclusion debriefing. AB - Approximately 16% of consumers are secluded during an inpatient admission. Despite the harmful psychological consequences resulting from the use of physical force, restraint, control, and temporary sequestration of therapeutic communication, there is little evidence of nursing practices to support consumers who are secluded. This paper will outline the findings from an action research project examining post-seclusion debriefing practice. A series of focus groups was undertaken with mental health nurses (MHN) and consumer consultants (CC) to investigate current practice, identify consumer/clinician preferences, and scope future practice possibilities. The findings suggest that CC need assistance in dealing with the negative feelings evoked by seclusion. MHN use a range of approaches for debriefing; however, these sometimes do not meet consumer preferences. MHN focus on explaining why seclusion happened and how to avoid it. While mitigation is a critical issue, CC want more emotional support from debriefing. MHN saw the possibility of using the debriefing to support consumers' transit from seclusion to high-dependency status and then to low-dependency status. Opinions raised regarding the potential for CC to have a role in debriefing were divided. The findings will be used to inform the development of a consumer debriefing training program for MHN and CC. PMID- 19290974 TI - Vulnerable populations and multicentred research. AB - There are many challenges facing researchers with projects requiring multicentred ethics approval. Achieving ethical approval at multiple sites, whether statewide or nationally, is a complex and time-consuming experience, compounded by the research process itself, as well as the recruitment of clinical sites and participants. Human ethics and research committees act as research gatekeepers and, as many research activities involve multiple applications and multiple approvals, can considerably delay the commencement of a project. A delay in ethics approval results in delays recruiting staff and participants, delays in the utilization of funding, and in delays regarding the progress and completion of projects. Such problems are additional problems for researchers of vulnerable populations, such as those in mental health or palliative care, where multicentred research is necessary to ensure the validity of the project itself. A current example of work that has required multicentred human ethics and research approval from around Australia is the establishment of the National Register of Antipsychotic Medication in Pregnancy. The guidelines for embarking on such a project with the requirement of multicentred ethical approval are described. Some of the issues, recommendations, and guidelines presented by the authors are taken from their experiences in establishing multicentred research projects. PMID- 19290975 TI - Psychiatry, mental health nurses, and invisible power: Exploring a perturbed relationship within contemporary mental health care. AB - Interpersonal relationships, although considered to be the cornerstone of therapeutic engagement, are replete with issues of power; yet, the concept of 'invisible power' within such formal mental health care relationships is seldom explored and/or critiqued in the literature. This paper involves an examination of power in the interpersonal relationship between the mental health nurse and the consumer. Issues of power are emphasized by drawing on examples from clinical experiences, each of which is then deconstructed as an analytical means to uncover the different layers of power. This examination highlights the existence of both obscure and seldomly acknowledged invisible manifestations of power that are inherent in psychiatry and interpersonal mental health nursing. It also identifies that there is an orthodoxy of formal mental health care that perhaps is best described as 'biopsychiatry' (or 'traditional psychiatry'). Within this are numerous serious speech acts and these provide the power for mental health practitioners to act in particular ways, to exercise control. The authors challenge this convention as the only viable discourse: a potentially viable alternative to the current of formal mental health care does exist and, most importantly, this alternative is less tied to the use of invisible power. PMID- 19290976 TI - Long-term homelessness in men with a psychosis: Limitation of services. AB - A proportion of persistently homeless men with psychosis are unable to function in the community, despite intensive outreach. The present study aimed to (i) develop a method to measure the number of homeless clients with a psychosis who are unable to maintain basic levels of function, despite the application of intensive community treatment; and (ii) determine if the proportion of these men had increased in an Australian inner urban setting over a 5-year period. Criteria were developed and validated to define this group. A review of records of clients engaged in homeless settings was conducted in order to compare the proportion of men with these features in 2000 and 2005. In 2005, 23% met the criterion of long term homelessness with poor function, despite intensive outreach, as compared with 17% in 2000 (P = 0.62). The 2005 cohort was more likely to have attempted intensive community psychiatric treatment (P = 0.04). We conclude that a proportion of homeless men with a psychosis in Australia do not achieve basic function, despite access to intensive outreach. The proportion of these men did not change between 2000 and 2005. These men require additional specialized services, including access to long-term supported housing. PMID- 19290977 TI - Testing and using Goldberg's General Health Questionnaire: Mental health in relation to home nursing, home help, and family care among older, care-dependent individuals. AB - The aims of this study were to test the Norwegian version of Goldberg's 30-item General Health Questionnaire (GHQ-30) in a group of older, care-dependent individuals living at home; to describe self-reported mental health; and to relate mental health to receiving home nursing, home help, and family care. A sample of 234 home nursing patients in Norway aged 75 years and older was interviewed. Mental state was assessed using the GHQ-30. Reliability and validity were calculated with Spearman's rank correlations, Cronbach's alpha coefficient, and Mann-Whitney U-test. The factor analysis was performed using the principal components analysis with varimax rotation and Kaiser normalization. Demographic characteristics and amounts of formal and family care were recorded, and descriptive statistics and stepwise multiple regression were used in the analyses. Cronbach's alpha coefficient for the GHQ was 0.92. The item-total correlations were generally acceptable. For items concerning depression and anxiety, the item-total correlations ranged from r(s)= 0.60 to 0.77. The factors extracted in the factor analysis explained 70% of the variance in the group. Females <85 years of age living in urban areas were associated with reduced mental health. There were no associations between general mental health and the amounts of formal and family care provided. PMID- 19290978 TI - Systematic screening for metabolic syndrome in consumers with severe mental illness. AB - The high prevalence of metabolic syndrome (MetS) in people with a mental illness has been reported recently in the literature. Gaps have emerged in the widespread use of systematic screening methods that identify this collection of critical risk factors for cardiac and metabolic disorders in people with severe mental illness. A sample (n = 103) of consumers with severe mental illness was screened for MetS using the Metabolic Syndrome Screening Tool and compared to a sample (n = 72) of consumers who were not receiving a systematic approach to screening for MetS. The results demonstrated ad hoc screening of consumers for MetS in the comparison group, potentially leaving patients at risk of cardiac and metabolic disorders being untreated. Mental health nurses are well placed to show leadership in the screening, treatment, and ongoing management of MetS in people with severe mental illness. A potential new speciality role entitled the 'cardiometabolic mental health nurse' is proposed as a means leading to improved outcomes for consumers who have both the complication of physical health problems and a severe mental illness. PMID- 19290979 TI - The journal in 2008. PMID- 19290980 TI - Validity of self-reported cardiovascular disease. PMID- 19290981 TI - Chronic lymphocytic leukaemia: current first-line therapy. AB - There is a major evolution in progress in the first-line therapy of chronic lymphocytic leukaemia. Several recent, large, clinical trials have documented superior outcomes with fludarabine-based therapy compared with treatment with alkylating agents. Monoclonal antibodies, especially rituximab, are establishing an important role for targeted treatment. It is expected that chemoimmunotherapy will become the preferred treatment for many patients in the near future. Specific challenges remain to be answered, however, especially the optimal treatment for the elderly, patients with autoimmune haemolysis and those with P53 deletions and mutations. PMID- 19290982 TI - Validity of self-reported cardiovascular disease events in comparison to medical record adjudication and a statewide hospital morbidity database: the AusDiab study. AB - Epidemiological studies often rely on self-reported cardiovascular disease (CVD) information, but this may be inaccurate. We investigated the accuracy of self reported CVD (myocardial infarction, stroke, coronary artery bypass surgery and coronary artery angioplasty) during the follow up of the Australian Diabetes, Obesity and Lifestyle (AusDiab) study. Self-reported CVD events, including the date of the event and hospital admission details, were collected with an interviewer-administered questionnaire. Of the 276 self-reported CVD events, 188 (68.1%) were verified by adjudication of medical records. Furthermore, linkage to the statewide Western Australian Hospital Morbidity Database (WAHMD) showed that CVD events were unlikely to be missed, with only 0.2% of those denying any CVD event being recorded as having had an event on the WAHMD. The adjudication of medical records was as accurate as record linkage to the WAHMD for validation of self-reported CVD, but combining the results from both methods of ascertainment improved CVD event identification. PMID- 19290983 TI - Anti-N-methyl-D-aspartate receptor antibody limbic encephalitis. AB - We report the case of a 57-year-old woman who developed acute psychiatric symptoms, behavioural disturbances, insomnia and dystonia resembling a catatonic state. During the course of her illness she developed hypoventilation and required monitoring in the intensive care unit. Her serum and cerebrospinal fluid showed antibodies to the NR1/NR2 heteromers of the N-methyl-D-aspartate receptor (NMDAR). Anti-NMDAR encephalitis is a severe form of autoimmune encephalitis, which has only recently been described in the published work. Most patients improve with immunosuppressive treatment. Raising awareness of this rare but increasingly reported condition is important, as it is responsive to treatment and potentially reversible. PMID- 19290984 TI - Butyrylcholinesterase: biomarker for exposure to organophosphorus insecticides. AB - Butyrylcholinesterase (BuChE) is routinely measured to assess exposure to or effects of organophosphorus insecticides (OP). As a biomarker, it can be used to clarify the relation between exposure to OP and health impairment. The interpretation of BuChE inhibition data, particularly of small changes in enzymatic activity, sometimes presents significant complexities. These complexities are presented in this short communication and the factors that influence the degree of BuChE inhibition are discussed. Despite the complexities of their interpretation, BuChE measurements remain a mainstay for the fast initial screening of exposure to OP; thus, they are a useful tool in the protection of humans, domestic animals and wildlife from overexposure to these toxic agents. PMID- 19290985 TI - Thrombolytic therapy for management of complicated catheter-related Candida albicans thrombophlebitis. AB - In immunocompromised patients, endovascular infection due to Candida albicans is associated with significant morbidity and mortality. Recommended management includes removal of any existing central venous catheter. Rarely, complications of endocarditis or infected mural thrombi may arise, with poorer clinical outcomes. For large endoluminal lesions, particularly of the great vessels or those that are intra-atrial, thrombolysis has been used in paediatric populations or before surgery for dissolution of infected thrombus. We describe the case of an adult patient with lung carcinoma who developed persisting candidaemia with a large endovascular fungal lesion adherent to the tip of a peripherally inserted central venous catheter. Local urokinase infusion enabled safe removal of the catheter without embolization. As an adjunct to antifungal therapy, local thrombolysis may play a contributory role in the management of central venous catheter-related candidal septic thrombosis. PMID- 19290986 TI - Aspergilloma. PMID- 19290987 TI - Pseudomembranous tracheobronchial aspergillosis. PMID- 19290988 TI - Chylothorax due to angioimmunoblastic T-cell lymphoma. PMID- 19290989 TI - Acute disseminated encephalomyelitis secondary to Mycoplasma pneumoniae. PMID- 19290990 TI - The impact of the measurement of uncertainty. PMID- 19290991 TI - Gadolinium-induced nephrogenic systemic fibrosis: communication breakdown among specialists or intuitive stroke of genius? PMID- 19290992 TI - The thrombotic thrombocytopenic purpura registry: a new national resource to inform patient care and medical research. PMID- 19290993 TI - Toll-like receptor 4 genetic diversity among pig populations. AB - The transmembrane glycoprotein encoded by the Toll-like receptor 4 gene (TLR4) acts as the transducing subunit of the lipopolysaccharide receptor complex of mammals, which is a major sensor of infections by Gram-negative bacteria. As variation in TLR4 may alter host immune response to lipopolysaccharide, the association between TLR4 polymorphisms and immune traits of the respiratory and gut systems has important implications for livestock. Here, a sequence dataset from 259 animals belonging to commercial and traditional European pig populations, consisting of 4305 bp of TLR4, including the full transcribed region, a portion of intron 2 and the putative promoter region, was used to explore genetic variation segregating at the TLR4 locus. We identified 34 single nucleotide polymorphisms, 17 in the coding sequence and 17 in the non-coding region. Five non-synonymous mutations clustered within, or in close proximity to, the hypervariable domain of exon 3. In agreement with studies in other mammals, a major exon 3 haplotype segregated at high frequency in the whole sample of 259 pigs, while variants carrying non-synonymous substitutions showed frequencies ranging between 0.6% and 8.7%. Although results on exon 3 provided suggestive evidence for purifying selection occurring at the porcine TLR4 gene, the analysis of both coding and non-coding regions highlighted the fact that demographic factors strongly influence the tests of departure from neutrality. The phylogenetic analysis of TLR4 identified three clusters of variation (ancestral, Asian, European), supporting the evidence of Asian introgression in European main breeds and the well documented history of pig breed domestication previously identified by mtDNA analysis. PMID- 19290994 TI - Noninvasive measurement of pH in platelet concentrates with a fiber optic fluorescence detector. AB - BACKGROUND: Stored platelets (PLTs) are metabolically active, resulting in a decrease of pH during storage. The pH of PLT concentrates (PCs) is recognized as a measure of quality, and pH limits are set by regulatory bodies. A pH sensor was built into a PLT storage container, and the feasibility of testing pH using a noninvasive fluorescent measurement method was evaluated. STUDY DESIGN AND METHODS: A citrated polyvinylchloride (PVC) PLT storage container with pH sensor insert was made and evaluated for biocompatibility during PLT storage and on pH reading accuracy, reproducibility, and durability. A noninvasive fluorescence reader was tested versus syringe-based sampling and subsequent measurement with a blood gas analyzer (BGA). The effect of interfering substances in plasma on the accuracy of this optical measurement was tested. Calibration and accuracy of the pH sensor were determined in both phosphate-buffered saline and in PCs. RESULTS: The citrated PVC storage container with pH sensor insert showed good storage properties for 300 mL of pooled buffy coat PLTs in plasma over 7 days. The pH sensor was easy to use and tracked pH(22) in the range of 6.2 to 7.8 over 11 days of storage. Accuracy in PCs was 0.08 pH units measured at 22 degrees C when calibrated against a BGA. CONCLUSION: The storage container with integrated pH sensor and noninvasive reader allows pH of PCs to be tracked over time in a noninvasive manner. PMID- 19290995 TI - WHO comparative evaluation of serologic assays for Chagas disease. AB - BACKGROUND: Evaluation of commercially available test kits for Chagas disease for use in blood bank screening is difficult due to a lack of large and well characterized specimen panels. This study presents a collaborative effort of Latin American blood centers and the World Health Organization (WHO) to establish such a panel. STUDY DESIGN: A total of 437 specimens, from 10 countries were collected and sent to the WHO Collaborating Center in Sao Paulo and used to evaluate 19 screening assays during 2001 through 2005. Specimens were assigned a positive or negative status based on concordant results in at least three of the four confirmatory assays (indirect immunofluorescence, Western blot, radioimmunoprecipitation assay, and recombinant immunoblot). RESULTS: Of the 437 specimens, 168 (39%) were characterized as positive, 262 (61%) were characterized as negative, and 7 (2%) were judged inconclusive and excluded from the analysis. Sensitivity and specificity varied considerably: 88 to 100 and 60 to 100 percent, respectively. Overall, enzyme immunoassays (EIAs) performed better than the other screening assays. Four EIAs had both parameters higher than 99 percent. Of the four confirmatory assays, only the RIPA gave a 100 percent agreement with the final serologic status of the specimens. CONCLUSION: The sensitivities and specificities of at least four of the commercially available EIAs for Chagas disease are probably high enough to justify their use for single-assay screening of blood donations. Our data suggest that the majority of commercially available indirect hemagglutination assays should not be used for blood donor screening and that the RIPA could be considered a gold standard for evaluating the performance of other assays. PMID- 19290997 TI - Digital pathology: a tool for 21st century neuropathology. AB - Digital pathology represents an electronic environment for performing pathologic analysis and managing the information associated with this activity. The technology to create and support digital pathology has largely developed over the last decade. The use of digital pathology tools is essential to adapt and lead in the rapidly changing environment of 21st century neuropathology. The utility of digital pathology has already been demonstrated by pathologists in several areas including consensus reviews, quality assurance (Q/A), tissue microarrays (TMAs), education and proficiency testing. These utilities notwithstanding, interface issues, storage and image formatting all present challenges to the integration of digital pathology into the neuropathology work environment. With continued technologic improvements, as well as the introduction of fluorescent side scanning and multispectral detection, future developments in digital pathology offer the promise of adding powerful analytic tools to the pathology work environment. The integration of digital pathology with biorepositories offers particular promise for neuropathologists engaged in tissue banking. The utilization of these tools will be essential for neuropathologists to continue as leaders in diagnostics, translational research and basic science in the 21st century. PMID- 19290998 TI - Application of telepathology for neuropathologic intraoperative consultations. AB - The demand for expert neuropathologic intraoperative diagnoses often exceeds the available supply and geographic distribution of neuropathology centers. Telepathology has therefore been implemented in recent years to meet this demand. Herein, we draw on our experience with 4 generations of telepathology systems over the past 8 years to discuss the design, initiation and maintenance of an effective telepathology service for neuropathologists, including when to change systems. In addition to workflow efficiency, unique advantages of telepathology include integration into other informatics modalities, quality assurance/quality control (QA/QC) maintenance and the potential for visual data to be readily available to clinicians. Given the improvements in technology and the multiple uses of telepathology, this method for delivering patient care will undoubtedly continue to grow over time. PMID- 19290999 TI - Internet and World Wide Web-based tools for neuropathology practice and education. AB - The Internet and the World Wide Web (www) serve as a source of information and a communication network. Together they form a so-called web or network that allows for transmission and dissemination of information in unprecedented speed, volume and detail. This article presents an overview of the current status of neuropathology content on the www. As well as considering the Internet as a resource for neuropathology practice, education and research, we also address the issue of quality assurance when evaluating Internet and www content. Four major categories of websites (archival, broker, news and blog) are discussed and resources relevant to neuropathology of each type are highlighted. We believe that our report and similar attempts can provide an opportunity to discuss appropriate and effective use of the Internet by the neuropathology community. PMID- 19291000 TI - A 32-year-old man with a fourth ventricular mass. AB - Lipoastrocytomas are rare and only four cases have been reported previously. Our case is in the fourth ventricle of a 32 year-old man.Many areas showed tumor cells with a signet-ring appearance and focally formed perivascular arrangements. Eosinophilic granular bodies were frequent. There was strong positive immunoreactivity to GFAP and S-100 protein, even in those vacuolated tumor cells.The proliferation index was less than 10%. Ultrastructural studies showed that the signet-ring cells had large lipid droplets as well as intermediate filaments. Ultrastructural features characteristic of ependymomas were not seen. These findings confirm that the vacuoles were lipid droplets in astrocytic tumor cells. The features noted are suggestive of a probable better prognosis in this variant of low-grade astrocytoma. PMID- 19291001 TI - A 33-year-old Chinese woman with a left frontal tumor. AB - Rhabdoid tumor cells are typically observed in atypical teratoid/rhabdoid tumor (AT/RT) but may also be seen in meningioma,glioma, melanoma, rhabdomyosarcoma and metastatic carcinoma.We present an astroblastoma with unusual rhabdoid features which is rarely described in the English literature. Apart from the rhabdoid tumor cells, all the histopathological features typical for astroblastoma are present in this case. These features include pseudopapillary arrangement, astroblastic pseudorosettes, perivascular hyalinization and calcifications, absence of fibrillary background and a pushing tumor border. The tumor cells display a multilineage immunohistochemical profile. In addition, diffuse and strong membranous and cytoplasmic dot-like pattern is appreciated with epithelial membrane antigen (EMA). The diagnosis of astroblastoma is also well supported by the age of presentation, anatomical location and radiological features of the tumor.We believe that on top of the above-mentioned unusual tumors with rhabdoid cells, astroblastoma should also be considered in the list of differential diagnosis. PMID- 19291002 TI - Posterior fossa tumor in a 12 year-old boy. AB - Cerebellar glioblastoma multiforme (GBM) has rarely been reported in children. We report a case of a 12-year-old child complaining of right upper limb tremor, loss of the normal capacity to modulate fine voluntary movements with right hand and headache, lasting for over a month. Radiological studies (CT and MRI) revealed a lesion of the right cerebellar hemisphere. The tumor was surgically excised and the histological examination revealed the presence of a GBM. The differential diagnosis of the lesions in the posterior fossa should include GBM. A gross total resection should be always attempted in order to achieve a better clinical outcome, although nearly all of these tumors recur. PMID- 19291003 TI - Posterior fossa tumor in a 2 year-old girl. AB - We report a case of a 2 year-old girl who presented with three weeks' history of deterioration of walking, then became unable to walk and later she developed projectile vomiting. Neurological examination revealed bilateral papilledema, nystagmus, and truncal ataxia with intention tremor. Radiological studies showed an enhancing mass in the posterior fossa extending from the cerebellum to the roof of the fourth ventricle. The tumor was diagnosed as an embryonal tumor with abundant neuropil and true rosettes (ETANTR). The tumor cells in the neuroblastic component were diffusely positive for synaptophysin and CD56, with scattered positive cells for glial fibrillary acidic protein. The true rosettes were only positive for vimentin. Ki67 showed high index (over 90%) in the true rosettes, while the neuroblastic areas were up to 15%. Our patient developed recurrent disease 6 months after resection and chemotherapy. ETANTR is a very rare aggressive embryonal CNS tumor that combines features of neuroblastoma and ependymoblastoma. We review the thirteen cases reported in the literatures. This case represents the second report of an ETANTR arising in the cerebellum. PMID- 19291004 TI - 72 year-old man with new seizure while dancing. AB - Metastasis of an extracranial malignant neoplasm to a meningioma is a rare event.We report a case of a 72-year-old man who presented abruptly with a grand mal seizure. Neuroradiological examination revealed an extraaxial lesion located in the posterior right frontal convexity with poorly defined boundaries. Histological and immunohistochemical examination showed that the lesion was a meningothelial meningioma harboring metastatic renal cell carcinoma.The MRI could be indicative but not specific of this type of lesion.Some cases of intracranial meningiomas containing metastatic carcinomas have been published, but to our knowledge only five cases of renal cell carcinoma metastasizing to a meningioma have been reported. Possible explanations for this type of "tumor in tumor" lesion are reviewed. PMID- 19291005 TI - A 15 year old boy with a posterior fossa tumor. AB - A 15 year old male presented with hydrocephalus from a tectal mass obstructing the cerebral aqueduct and upper fourth ventricle. The solid-cystic partly enhancing mass proved to be a pigmented pleomorphic xanthoastrocytoma, the third such example reported. The lesion revealed typical features of a PXA with the unusual addition of intracytoplasmic melanin in select lesional cells. Melanin pigment production is uncommon in glial tumors and of uncertain significance. The present case is recurrence-free one year post-operatively. PMID- 19291006 TI - Conservation and diversification of the symmetry developmental program among close relatives of snapdragon with divergent floral morphologies. AB - Multiple evolutionary shifts in floral symmetry and stamen number have occurred in the snapdragon (Antirrhinum majus) family Veronicaceae. In Mohavea, Veronica and Gratiola there have been independent evolutionary reductions in stamen number and modifications to corolla shape. It is hypothesized that changes in the regulation of homologs of snapdragon dorsal flower identity genes CYCLOIDEA (CYC) and RADIALIS (RAD) underlie these floral transitions. CYC-like and RAD-like genes from Veronica montana and Gratiola officinalis were cloned and sequenced, compared with homologs from other Veronicaceae species using phylogenetic analysis, and their expression was investigated by reverse transcriptase polymerase chain reaction (RT-PCR) and in situ hybridization. VmCYC1, GoCYC1, GoCYC2 and RAD-like genes are expressed exclusively in the dorsal region of floral meristems and developing flowers. Their expression patterns do not correlate with patterns of stamen arrest. VmCYC2 and GoCYC3 are expressed in both vegetative and floral tissues, with VmCYC2 being most abundant in all regions of the floral meristem and all petals. These results support conservation of the floral symmetry gene network for Veronicaceae RAD-like and some CYC-like paralogs, suggest regulatory evolution of other CYC-like genes following gene duplication and implicate different genetic mechanisms underlying dorsal versus ventral stamen abortion within Veronica and Gratiola. PMID- 19291007 TI - Characterization of a MADS FLOWERING LOCUS C-LIKE (MFL) sequence in Cichorium intybus: a comparative study of CiMFL and AtFLC reveals homologies and divergences in gene function. AB - In Arabidopsis thaliana, the ability to flower is mainly related to a floral repressor, FLOWERING LOCUS C (FLC), which is regulated through the vernalization pathway. The genes controlling the vernalization pathway seem to be only partially conserved in dicots other than the Brassicaceae. Cichorium intybus (chicory) is a biennial species belonging to the Asteraceae family, and it shows an obligate vernalization requirement for flowering. Cichorium intybus MADS (MCM1, Agamous, Deficiens, SRF) FLC-like (CiMFL) sequences were isolated in C. intybus by RT-PCR and their expression patterns characterized during plant development and in response to vernalization. The biological function of CiMFL was analysed by complementation of A. thaliana FRIGIDA (AtFRI);flc3. Resetting of MFL expression after vernalization was analysed during microsporogenesis. Before vernalization, CiMFL is mainly expressed in the axils of young leaves. Vernalization induced CiMFL down-regulation under a long-day photoperiod but not under a short-day photoperiod. Furthermore, together with a decrease in CiMFL transcripts, cold conditions induced changes in the morphology of the shoot apical meristem and in the transition to flowering. The biological function of CiMFL was found not to be conserved. Our results show that the regulation of CiMFL expression in time and space and in relation to environmental conditions is only partially conserved with respect to FLC isolated from A. thaliana. A model for flowering repression by CiMFL is proposed. PMID- 19291008 TI - Quantitative genetic analysis of biomass and wood chemistry of Populus under different nitrogen levels. AB - The genetic control of carbon allocation and partitioning in woody perennial plants is poorly understood despite its importance for carbon sequestration, biofuels and other wood-based industries. It is also unclear how environmental cues, such as nitrogen availability, impact the genes that regulate growth, biomass allocation and wood composition in trees. We phenotyped 396 clonally replicated genotypes of an interspecific pseudo-backcross pedigree of Populus for wood composition and biomass traits in above- and below-ground organs. The loci that regulate growth, carbon allocation and partitioning under two nitrogen conditions were identified, defining the contribution of environmental cues to their genetic control. Sixty-three quantitative trait loci were identified for the 20 traits analyzed. The majority of quantitative trait loci are specific to one of the two nitrogen treatments, demonstrating significant nitrogen-dependent genetic control. A highly significant genetic correlation was observed between plant growth and lignin/cellulose composition, and quantitative trait loci co localization identified the genomic position of potential pleiotropic regulators. Pleiotropic loci linking higher growth rates to wood with less lignin are excellent targets to engineer tree germplasm improved for pulp, paper and cellulosic ethanol production. The causative genes are being identified with a genetical genomics approach. PMID- 19291009 TI - Corticotropin-releasing factor-1 receptor antagonists decrease heroin self administration in long- but not short-access rats. AB - Dysregulation of the stress-related corticotropin-releasing factor (CRF) system has been implicated in the development of drug dependence. The present study examined the effects of administering CRF type 1 (CRF(1)) receptor antagonists on heroin self-administration in animals allowed short (1 hour) or long (8-12 hours) access to intravenous heroin self-administration sessions. The nonpeptide CRF(1) antagonists MJL-1-109-2 (1 hour versus 8 hours access) or R121919 (1 hour versus 12 hours access) were systemically injected in both short- and long-access rats. MJL-1-109-2 (10 mg/kg) and R121919 (10 and 20 mg/kg) reduced heroin self administration in long-access animals without altering heroin intake in short access animals. Both MJL-1-109-2 and R121919 decreased first-hour intravenous heroin self-administration selectively in long-access rats, with R121919 decreasing cumulative heroin intake across the 12-hour session. The results demonstrate that blockade of the CRF-CRF(1) receptor system attenuates the increased heroin intake of rats with extended access to the drug. PMID- 19291010 TI - Attentional bias in alcohol-dependent patients: the role of chronicity and executive functioning. AB - It has been suggested that the attention towards alcohol-related stimuli increases with the duration of drinking and alcohol dependence. The present study aimed to assess whether an attentional bias was present in detoxified alcohol dependent patients, and if the magnitude of the attentional bias depended on the subject's drinking history and variables of executive functioning. Attentional bias was assessed in 30 alcohol-dependent patients using a visual dot-probe task with a picture presentation time of 50 ms. In addition, patients completed a variety of different cognitive tasks such as attention, continuous performance, working memory, set shifting and inhibitory control tests. Based on correlation analysis we split the patient sample on the median with regard to the duration of alcohol dependence and our results indicated a significant attentional bias towards alcohol-associated pictures in patients dependent for less than 9 years, but not in patients with a longer duration of dependence. The two patient samples differed significantly with regard to attention and working memory functioning with patients who were dependent for more than 9 years showing a greater impairment. When impairment of attention and working memory were controlled for, the group differences in attentional bias were no longer significant. Our results indicate that differences with regard to drinking-related variables as well as cognitive functioning seem to modulate attentional bias and need to be taken into account in models of drinking maintenance. PMID- 19291011 TI - Retention rate and side effects in a prospective trial on hepatitis C treatment with pegylated interferon alpha-2a and ribavirin in opioid-dependent patients. AB - Hepatitis C viral (HCV) infection is present in 30 to 98% of intravenous drug users. Intravenous substance abuse represents the main route of HCV transmission in industrialized countries. A multi-centre, randomized, controlled, prospective study assessed sustained virological response (SVR), adverse events such as depressive episodes and retention rate of HCV treatment in opioid-dependent patients. Stabilized, opioid-dependent patients with chronic HCV infection (genotype 2 or 3) received pegylated interferon alpha-2a in combination with ribavirin 800 mg/day (Group A) or 400 mg/day (Group B). Participants were randomized, blocked and stratified by genotype and viral load. A standardized psychiatric assessment, Beck Depression Inventory (BDI) and Van Zerssen's list of complaints were administered at each study visit. In 31 months, 300 opioid dependent patients were screened; 190 (63.3%) were hepatitis C antibody positive. According to study protocol, out of 75 'potential-to-treat' patients with genotype 2 or 3, 17 stable patients (22.6%) were included in the study. All participants completed the study. Significant haemoglobin decreases occurred in both Groups A (P = 0.001) and B (P = 0.011). All the patients had an end-of treatment (week 24) HCV RNA negativity. Fifteen (88.2%) achieved SVR at week 48. Overall, 52.9% developed depressive symptoms during treatment. Because of the prompt initiation of antidepressant medication at first appearance of depressive symptoms, no severe depressive episodes occurred. Our data show a high retention rate and reliability, and good viral response for both treatments. Hepatitis C treatment in stable opioid-dependent patients was efficacious, suggesting that addiction clinics can offer antiviral therapy in combination with agonistic treatment as part of multi-disciplinary treatment. PMID- 19291013 TI - Myocardial metastasis in patient with cholangiocarcinoma. AB - Myocardium is a rare site for metastasis. We present here the case of myocardial metastasis from cholangiocarcinoma. PMID- 19291014 TI - The many faces of strain. PMID- 19291015 TI - Non-Doppler two dimensional strain imaging for evaluation of coronary artery disease. AB - Over the recent years, strain echocardiography has emerged as a quantitative technique for the evaluation of global and segmental cardiac function. Strain is a measure of deformation, expressed as a percent change in a segment's length compared to its predeformation length. Strain rate (SR) is the local rate of deformation or strain per unit time. Recently non-Doppler two dimensional strain imaging has been developed. This technique is based on tracking ultrasonic speckles from the two dimensional echocardiographic images. These speckles are followed over a number of successive frames, and myocardial velocity is calculated by measuring frame-to-frame changes. This technique is independent of the Doppler angle of incidence and allows measurement of several vectors of strain within myocardial tissue. Non-Doppler strain is a powerful tool, enabling detection of subtle abnormalities in myocardial function. Current evidence shows that non-Doppler strain imaging may allow identification of the early changes that occur with ischemic insult to the myocardium. It may also provide a tool for identification of scarred, non-viable myocardium, with similar accuracy to that of cardiac MRI. Non-Doppler strain imaging is likely to become a standard tool in the evaluation of patients with ischemic heart disease. PMID- 19291016 TI - The value of 2D strain imaging during stress testing. AB - Stress echocardiography is increasingly used but its major limitation is the subjective interpretation of wall motion changes requiring experience. Speckle tracking enables simultaneous evaluation of radial, longitudinal, and circumferential myocardial deformation. Recently, two-dimensional (2D) strain has been found to be as reliable as sonomicrometry for the assessment of left ventricular (LV) regional function. In the presence of inducible ischemia, longitudinal and circumferential abnormalities preceed the decrease in radial deformation. Optimal cutoffs have been obtained from 2D strain rate (SR) at peak dobutamine stress to predict coronary artery disease. However, 2D strain rate does not yet provide incremental accuracy to visual interpretation by experts. Speckle tracking strain could be useful to better identify contractile reverse and biphasic response of viable myocardium but there are not yet clinical studies published in this setting. Preliminary results suggest that 2D strain obtained during exercise could be useful in asymptomatic patients with severe aortic stenosis or organic mitral regurgitation (MR). In conclusion, the reliability and clinical importance of 2D strain during stress will be specified by further investigations. PMID- 19291017 TI - The role of speckle tracking strain imaging in cardiac pacing. AB - In recent years, concerns have been raised about possible harmful effects of long term right ventricular (RV) apical pacing. These detrimental effects may be related to changes in left ventricular (LV) mechanics during RV apical pacing. As a consequence, alternative RV pacing sites have been proposed, and in selected patients an upgrade from RV to biventricular pacing may be considered. Novel two dimensional (2D) speckle tracking strain imaging allows detailed evaluation of LV mechanics, including LV mechanical dyssynchrony, LV strain and LV torsion. In this review, the role of speckle tracking strain imaging in the evaluation of LV function in patients with RV apical pacing will be reviewed. The effects of RV apical pacing on LV mechanical dyssynchrony, LV strain and LV torsion will be discussed. In addition, the role of speckle tracking strain imaging in the selection of the optimal (alternative) RV pacing site and in the selection of patients who may benefit from an upgrade from RV apical pacing to biventricular pacing will be reviewed. PMID- 19291018 TI - Speckle tracking in the evaluation of left ventricular dyssynchrony. AB - A number of echocardiographic techniques have been introduced to determine left ventricular dyssynchrony (LVD) and to improve selection of patients for CRT. During the last years tissue Doppler imaging (TDI) has been used as the most preferred technique to quantify LVD, but results with nonresponder rates below 30% have been shown only in small studies based on high experience. Angle of incidence dependency, noise, artifacts, and tethering motion of adjacent segments are the main limitations of TDI influencing selection of patients for CRT. Although strain TDI is not affected by translation or tethering, accurate measurement of regional strain is also limited. Two-dimensional (2D) strain imaging based on novel speckle tracking echocardiography (STE) is a relatively new tool to define regional myocardial strain and to quantify dyssynchrony based on a more robust technique and avoiding angle of incidence. Current studies are promising to use strain or vector velocity imaging derived from STE for qualitative and quantitative assessment of LVD and follow-up studies as well. If one compare different types of strain components at present, radial strain imaging seems to be the most promising technique to determine LVD and to predict positive response to CRT. Furthermore, STE offers an insight into rotational mechanics of the dyssynchronous ventricle. Although clinical studies using 2D strain have analyzed LVD related to various conditions, measures are based on a 2D data set. Three-dimensional strain imaging, based on speckle tracking will probably open a new door to assess patients with heart failure and LVD. PMID- 19291019 TI - Strain and strain rate imaging in cardiomyopathy. AB - The most common indication for an echocardiogram is for the assessment of left ventricular (LV) function and, in the evaluation of cardiomyopathy (CM), this becomes even more important. However, conventional echocardiographic measures of ventricular function are insensitive at detecting subtle perturbations in contractility. In patients with CM, the ability to detect abnormalities early in the course of the disease to establish a diagnosis can be critical and often may influence specific treatments administered as well as establish important prognostic information. Technologic advances in echocardiographic imaging during the last decade now allow for the measurement of LV strain and strain rate (SR) imaging. Strain and SR imaging allow for a more precise characterization of the mechanics of myocardial contraction and relaxation (deformation imaging) and emerging data are establishing the use of these techniques in a variety of different cardiomyopathic conditions. After establishing a common understanding of strain imaging as well as defining the methods by which these measures can be incorporated into an echocardiographic examination, we will review the accumulating information illustrating the great promise that this imaging modality has in the care of patients with CM. This review will focus on the role of strain and SR imaging in CM. PMID- 19291020 TI - Two-dimensional, non-Doppler strain imaging during anesthesia and cardiac surgery. AB - Transesophageal echochardiography (TEE) has become an essential intraoperative monitor during general anesthesia for cardiac surgical procedures. In clinical practice, ventricular function is visually evaluated using gray scale and Doppler modes, despite the fact that subjective interpretation is influenced by level of experience and training. Echocardiographic strain imaging measures cardiac deformation and provides objective quantification of regional myocardial function. Non-Doppler strain, which is derived by tracking speckles from two dimensional (2D) images, bypasses the limitations of Doppler-based strain measurements and evaluates the complex myocardial deformation along three dimensions. As a result, longitudinal shortening, circumferential thinning and radial thickening can be quantified using standard midesophageal and transgastric views, being acquired during a comprehensive TEE examination. Once non-Doppler strain becomes available on "real time," it will have the potential to become a valuable tool for detection of ischemia on the regional level and objective quantification of global ventricular function. PMID- 19291021 TI - Re: Echocardiographic left ventricular mass in a multiethnic Southeast Asian population: proposed new gender and age-specific norms. PMID- 19291023 TI - Identification of sulfate-reducing bacteria in methylmercury-contaminated mine tailings by analysis of SSU rRNA genes. AB - Sulfate-reducing bacteria (SRB) are often used in bioremediation of acid mine drainage because microbial sulfate reduction increases pH and produces sulfide that binds with metals. Mercury methylation has also been linked with sulfate reduction. Previous geochemical analysis indicated the occurrence of sulfate reduction in mine tailings, but no molecular characterization of the mine tailings-associated microbial community has determined which SRB are present. This study characterizes the bacterial communities of two geochemically contrasting, high-methylmercury mine tailing environments, with emphasis on SRB, by analyzing small subunit (SSU) rRNA genes present in the tailings sediments and in enrichment cultures inoculated with tailings. Novel Deltaproteobacteria and Firmicutes-related sequences were detected in both the pH-neutral gold mine tailings and the acidic high-sulfide base-metal tailings. At the subphylum level, the SRB communities differed between sites, suggesting that the community structure was dependent on local geochemistry. Clones obtained from the gold tailings and enrichment cultures were more similar to previously cultured isolates whereas clones from acidic tailings were more closely related to uncultured lineages identified from other acidic sediments worldwide. This study provides new insights into the novelty and diversity of bacteria colonizing mine tailings, and identifies specific organisms that warrant further investigation with regard to their roles in mercury methylation and sulfur cycling in these environments. PMID- 19291024 TI - Molecular methods to detect and monitor dissimilatory arsenate-respiring bacteria (DARB) in sediments. AB - Dissimilatory arsenate-respiring bacteria (DARB) reduce arsenate to arsenite and may play a significant role in arsenic mobilization in aquifers and anoxic sediments. Many studies have been conducted with pure cultures of DARB to understand their involvement in arsenic contamination. However, few studies have examined uncultured DARB in the environment. In order to investigate uncultured DARB in anoxic sediments, genes encoding arsenate respiratory reductases (arr) were targeted as a genetic marker. Degenerate primers for the alpha-subunit of arr genes were designed and used with PCR amplification to detect uncultured DARB in the sediments collected from three stations (upper, mid and lower bay) in the Chesapeake Bay. Phylogenetic analysis of putative arrA genes revealed the diversity of DARB with distinct community structures at each of the three stations. Arsenate reduction in sediment communities was confirmed using enrichment cultures established with sediment samples from the upper bay. In addition, terminal restriction fragment length polymorphism analysis of the putative arrA genes showed changes in the community structure of DARB in the enrichment cultures while reducing arsenate. This was also confirmed by cloning and sequence analysis of the arrA genes obtained from the enrichment cultures. Thus, we were able to detect diverse uncultured DARB in sediments, as well as to describe changes in DARB community structure during arsenic reduction in anoxic environments. PMID- 19291025 TI - Screening tests for free radical scavengers on cutaneous cultured cells. AB - Synopsis Numerous studies discuss the important development of free radicals in human skin after exposure to UV irradiation. They suggest that these reactive molecules are responsible for sun-accelerated cutaneous aging. We reproduced two types of radical agression on cutaneous human cell cultures. The first brings into action oxygen radicals generated by hypoxanthine-xanthine oxidase system and the second involves the use of UV radiation. These complementary models may be used as screening methods for antioxidant molecule research because they allow the distinction between molecules with filtering properties and free radical scavengers. The possibility of introducing molecules at different stages (before, during and after exposure to agression) permits the study of the molecular mecanism. These models were tested on silymarin, catechin, ascorbic acid and alpha tocopherol. PMID- 19291026 TI - Sensitivity to perfumes and preservatives in patients with contact dermatitis. AB - Synopsis Patients with contact dermatitis were patch tested for sensitivity to perfumes (perfume mixture, balsam of Peru) and to preservatives (formaldehyde). The percentages of sensitivity in Greek patients with contact dermatitis were 8.1% for the perfume mixture, 3.7% for the balsam of Peru and 2.3% for formaldehyde. It is notable that 1.4% of the patients demonstrated simultaneous sensitivity to the perfume mixture and to balsam of Peru, while no case of simultaneous sensitivity occurred for formaldehyde and the two other compounds. Factors such as sex, age, occupation and anatomic site of the appearance of dermatitis in patients were taken into account and related to the percentages of sensitivity. Comparing with the literature we have concluded that the frequency of appearance of sensitivity by Greek patients is not significantly different from the average observed for patients in Northern or Southern Europe. PMID- 19291027 TI - Studies of factors affecting light scattering by individual human hair fibres. AB - Synopsis An automated goniophotometer has been built to investigate the scattering of light by different types of singly mounted human hair fibres using white light and laser illumination. The effects that varying hair colour and treatment (gold coating, ultraviolet irradiation and dyeing) have on the specular reflectance and on the half-peak-height width (omega) have been studied. The polarization of light has been used to discriminate between light reflected from the front surface and light scattered internally by the hair. It has been found that the analysis of results from individual hair fibres has been useful in understanding the light scattering mechanisms which occur in human hair. PMID- 19291028 TI - Efficiency and usability of silicone surfactants in emulsions. AB - Synopsis The critical micelle contration (CMC) of silicone surfactants was determined to provide their optimal efficiency in emulsion systems. The lowest CMC value was found for the amphionic polysiloxane polyorganobetaine copolymer, which is undoubtedly the consequence of electrostatic repulsion preventing the aggregation of ionized molecules. Also the nonionic surfactants (polydimethyl siloxane polyethers) showed some differences: the surfactant with polyethylene oxide side chain is more effective compared to the polyethylene-polypropylene oxide side chain. The results of miscibility testing of silicone surfactants with common components of emulsions and creams confirm that not only silicone oils, but also silicone surfactants, should be considered as both explicitly hydrophobic and non lipophilic. The stability of emulsions containing silicone surfactants prepared by cold emulsification supported the incorporation of polyelectrolyte sodium carboxymethylcellulose, which increased viscosity of the continuous phase and prevented coalescence. The physical stability testing of prepared emulsions confirms that the used methods - centrifugation, particle size analysis and the observation of ageing at room (20-25 degrees C) and elevated temperature (40 degrees C) - give comparable results. PMID- 19291029 TI - Transparent formulations of a liposoluble sunscreen agent in an aqueous medium. AB - Synopsis Transparent formulations of a liposoluble sunscreen in an aqueous medium were studied, taking into account the different chemicals of a selected system situated in the vertices of a regular tetrahedron: sunscreen agent, co-solvent, (surfactants +/- mineral oil) mixture (in the tetrahedron base) and water as the fourth component. Several compositions were selected at optimized component ratios after which water was added progressively. Clear gel and liquid compositions were obtained yielding a sunscreen agent that it suitable for conventional use. The physico-chemical properties of some compositions were assessed in terms of water/sunscreen agent content. PMID- 19291030 TI - The skin microflora and the formation of human axillary odour. AB - Synopsis We have examined the relationship between human axillary skin microflora and underarm odour (UAO), in particular, the ability of cutaneous bacteria to transform steroids. A study was made of bacterial population density and odour intensity of the axillae of 34 normal male subjects. There was a statistically significant association between population density of aerobic coryneform bacteria and UAO intensity. No associations could be found between population densities of staphylococci, micrococci or propionibacteria and UAO intensity. An in vitro model for formation of UAO was developed, and used to test individual bacterial isolates. Only aerobic coryneforms could produce axillary odour in vitro, most notably C. xerosis. Many aerobic coryneforms could transform testosterone, the principal metabolites being 5alpha- and 5beta-DHT, androstenedione, and 5alpha- and 5beta-androstanedione. UAO positive coryneforms were more metabolically active than UAO negative bacteria. Micrococci also transformed testosterone to androstenedione, whilst staphylococci and propionibacteria could not metabolize it. A hypothesis for the role of aerobic coryneforms in the formation of human axillary odour is discussed. PMID- 19291031 TI - Studies on the interaction of p-phenylene diamine with phospholipids. AB - Synopsis In vitro studies on the interaction of phospholipids with p-phenylene diamine, a major component of hair dyes, were undertaken. Thin layer and molecular sieving chromatography along with phosphorus estimation and spectral characterization suggested the possibility of p-phenylene diamine and its biotransformation products combining with lipophilic biomolecules. PMID- 19291032 TI - Emulsification by the phase inversion temperature method: the role of self bodying agents and the influence of oil polarity. AB - Synopsis Oil-in-water emulsions stabilized with nonionic emulsifiers change to water-in-oil emulsions as the temperature rises when the hydrophilic and lipophilic properties of the mixed emulsifier are just balanced. Preparation above the phase inversion temperature followed by rapid cooling yields emulsions that exhibit very fine droplet size and extreme long-term stability. Cosmetic emulsions were prepared by this phase inversion temperature (PIT) method using typical raw materials such as polar oils, e.g. decyl oleate, 2-octyl dodecanol or isopropyl myristate, and nonionic emulsifiers, e.g. ceteareth-12 or polyoxyethylene eicosyl/docosyl ether combined with cetostearyl alcohol as a co emulsifier. The phase inversion temperature was measured as a function of the oil polarity and the concentration of mixed emulsifier. The relationship between phase inversion temperature, droplet size and emulsion stability was investigated. In addition, self-bodying agents such as cetostearyl alcohol or monoglycerides were added to these thin, fine disperse emulsions to adjust the consistency. The influence of these ingredients on phase inversion temperature, droplet size, yield value and emulsion stability was studied. PMID- 19291033 TI - Measurement of beard hair removal efficacy using image analysis. AB - Synopsis Using an image analyzer, a method was developed for measuring differences in the beard hair removal efficacy of shaving systems. The method involves the collection of shaved whiskers from three panels of shavers. The panelists use two different shaving products on the two sides of their faces which they alternate from day to day. The lengths or projection areas of about 200 whiskers from each shave is then measured using an electronic image analyzer. By comparing the mean values of the whisker lengths (areas), differences in hair removal efficacies less than 1% can be measured at statistically significant levels. The protocol allows the determination of differences in both the first and second cuts of hair lengths, when twin bladed razors are used. PMID- 19291034 TI - Influence of hazelnut oil phospholipids on the skin moisturizing effect of a cosmetic emulsion. AB - Synopsis Previous studies have shown significant differences between the moisturizing effects and skin tolerances of virgin and refined vegetable oils when incorporated in cosmetic emulsions. They have also shown significantly greater and longer lasting moisturizing potential for a cosmetic emulsion containing virgin vegetable oils when compared with refined oils in the same emulsion. Results were obtained with sweet almond and hazelnut oils. Hazelnut oil has now been analysed to show the effect of refining. The phospholipid content of the oil decreased from 286 ppm in virgin oil to traces in refined oil. These oils were added at 10% to a cosmetic emulsion applied twice a day for 28 days to the skin of 56 volunteers and the changes in moisturizing effect measured by means of the capacitance method (corneometer CM 240 Khazaka). Measurements were made under conditions of controlled temperature and humidity on the first day of application and repeated after 21 and 28 days of treatment. A significant increase in the moisturizing effect of the emulsion containing virgin hazelnut oil was found and this was itself significantly greater than the emulsion containing refined oil. Both emulsions had a greater moisturizing effect when compared with the control emulsion and the untreated skin. Phospholipids were isolated by dialysis from virgin oil and added to the refined oil (234 ppm). The moisturizing effect obtained with the emulsion containing the enriched refined oil was shown to be the same as that obtained with the virgin oil emulsion. PMID- 19291035 TI - A new methodology for testing chemicals and drugs on cell activity. AB - Synopsis A new method for testing the effects of cosmetics and various chemicals on cell activity is described. It is based on the study of mitochondria compartments and combines the use of fluorescent probes which are accumulated in a potential-dependent (Rhodamine 123) or potential-independent (Nonyl Acridine Orange) manner with flow cytometric analysis. Measured fluorescences allow the determination of a new parameter indicating mitochondria efficiency. This new methodology for the evaluation of cell activity has been applied in three different situations: during cell ageing, during cell cycle and when cells are in the presence of drugs. PMID- 19291036 TI - Dowicil 200 Stability test: chemical and microbiological studies. AB - Synopsis The stability of the formaldehyde-releaser preservative Dowicil 200 has been investigated. The studies were carried out on cosmetic emulsions preserved with different concentrations of Dowicil 200, stored in the dark at 45 degrees C or 25 degrees C. The degree of microbiological activity was tested with respect to S. aureus, P. aeruginosa, E. coli and A. niger using the agar diffusion and challenge tests. The chemical stability was evaluated by HPLC in the reverse phase mode. A significant preservative instability was observed. PMID- 19291037 TI - Preliminary Communication: Treatment of irritant contact dermatitis in workmen with perfluoropolymethylisopropyl ether (Fomblin HC). AB - Synopsis Non-allergic contact dermatitis is a major dermatological problem for the workmen of an important factory which manufactures industrial machines (SAIMP SpA) located at Padua. Such dermatitis is due to the presence in the environment of paints, solvents, surfactants, lubricants and metal powders (steel and cast iron). As medical treatment, the factory doctor practises the use of common barrier creams which do not give completely satisfactory results. To cope with this problem, some workmen were treated with preparations containing Fomblin HC. The highly successful results of this new type of treatment are reported: among other findings this study shows, through experimental evidence, that Fomblin HC has a protective effect not only against water soluble substances but also against oil soluble substances (in fact, the latter are probably the main cause of contact dermatitis contracted by these workmen). PMID- 19291038 TI - [Not Available]. AB - Synopsis The main purpose of this article is to define multiple emulsions, their preparation (new materials and manufacturing processes) methods of characterisation and analysis, and their applications in cosmetology and pharmacy. The second part is devoted to a presentation of the investigation in our laboratory into the optimal manufacturing process to provide stable formulations. The study of release mechanisms from such formulations mainly shows a breakdown of multiple droplets which could be controlled by variation of solute concentration. PMID- 19291039 TI - Dental plaque - associated infections and antibacterial oral hygiene products. AB - Synopsis Dental plaque accumulates on hard non-shedding surfaces such as teeth, dentures and orthodontic appliances. This accumulation is facilitated by the absence of adequate oral hygiene procedures. The term 'plaque' describes a mass of microorganisms embedded in an organic matrix of host and microbial origin. In addition to the aesthetic desirability of 'clean teeth, healthy gums and fresh breath' associated with the absence of plaque, obvious consequences of the presence of plaque include tooth decay (dental caries), gingivitis and periodontal (gum) disease and denture associated problems. Thus the prevention of plaque formation, the reduction of plaque accumulation and the effective removal of plaque are considerations of the cosmetic and health professions alike. There are many oral hygiene products available to the general public - toothpastes, mouthwashes, denture cleaners, and, more recently, chewing gums and novel mouthwashes. Several of these products have antimicrobial components. This paper reviews the microbiology of plaque and plaque associated problems, and surveys the type of products currently available for maintenance of good oral hygiene. Potential areas for future development are also explored. PMID- 19291040 TI - Viscosity regulation by surfactants. AB - Synopsis The viscosity of cosmetic emulsions is of great practical importance and related to their skin feel. To achieve a desirable and constant viscosity is not an easy task because numerous factors must be considered. Based on theoretical considerations a new surfactant, steareth-5-stearate, has been developed. It can be shown that its use, together with a second more hydrophilic surfactant, results in flowable emulsions, the viscosity of which remain stable under normal conditions. PMID- 19291041 TI - Prophylaxis of Striae gravidarum with a topical formulation. A double blind trial. AB - Synopsis A prophylactic antistriae cream (Centella asiatica extract, alpha tocopherol, and collagen-elastin hydrolisates) was assessed by a double blind trial in 80 pregnant women. In the placebo group 22 women (56%) presented striae, whereas in the treated group only 14 women (34%) developed striae in this pregnancy; this difference was significant (p < 0.05; chi(2) test). An arbitrary score was designed to assess the intensity of striae (from 0 to 3); this score was 1.42 (sd 0.5) in the treated group and 2.13 (sd 1.32) in the placebo group and this difference was also significant (p= 0.014; Mann-Whitney test). In women with a history of striae during puberty, the active cream induced a significant absolute prevention in 89% of the cases whereas in the placebo group all the women developed striae (p= 0.00014; chi(2) test). PMID- 19291042 TI - HPLC determination of thioglycolic acid and other aliphatic thiols in cosmetic formulations using ethacrynic acid as precolumn derivatization reagent. AB - Synopsis Limits imposed on the usage of thioglycolic acid, its salts and esters in cosmetic formulations require selective and sensitive analytical methods for their determination. In this study a convenient and reliable method based on high performance liquid chromatography (HPLC) has been developed. The method involves a precolumn reaction of the thiol compounds with ethacrynic acid to give thiol adducts which can be separated by reversed-phase liquid chromatography and detected at lambda= 273 nm. The derivatization reaction proved to be quantitative under mild conditions (20 min at pH 7.4 and room temperature) and the chromatographic conditions allowed thioglycolic acid, glyceryl monothioglycolate, thiolactic acid, and thioglycolic acid ethyl ester to be discriminated. The proposed method was successfully applied to the analysis of commercial cosmetic formulations containing thioglycolic acid and glyceryl monothioglycolate, fulfilling the requirements of a general and selective method for the analysis of aliphatic thiols in cosmetics. PMID- 19291043 TI - Methods for assessment of cutaneous ageing. AB - Summary Quantitative assessment of skin ageing is necessary for the evaluation of age modifying treatments. The particular method used must depend on the target of the intended treatment. It should also clearly distinguish between the degree of chronic environmentally-induced trauma to the skin (predominantly photoageing) and the intrinsic ageing process. However, it is clear that no one measure will be adequate to define the extent of cutaneous ageing. Broadly speaking, the techniques available are either invasive or non-invasive. Both have their uses. For example, topically applied retinoic acid 0.05% has been shown to increase epidermal thickness and decrease stratum corneum replacement time, using an invasive technique to determine the value for the first and a non-invasive method to evaluate the second. Virtually all aspects of skin structure and function change with age. However, because of the importance with regard to appearance, some measure of solar elastotic degenerative change should be made. Profilometry techniques and ultrasound determination of skin thickness, as well as measurement of skin colour, may all have their use for this purpose. PMID- 19291044 TI - Direct contact membrane method for evaluating preservative efficacy in solid cosmetics. AB - Synopsis A method was developed for evaluating in-product preservative efficacy of solid cosmetics. Microbes (10(5)-10(6) colony-forming units) resting on membrane filters were placed in direct contact with the surface of pressed eye shadows at room temperature in a moist chamber. At daily intervals, or as appropriate, the filters were removed and viable counts were determined by pour plates of modified letheen agar. Linear regression analysis was used to evaluate preservative efficacy. Decimal reduction times (D values) of 1.2-1.7 days were obtained for Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 8739 on in-house pressed eye shadows preserved with parabens and Germall 115. An average D value of 1.7 was also obtained for P. aeruginosa and S. aureus on a commercially pressed eye shadow containing the same preservatives as the in-house formulation. However, this commercial product was either totally ineffective or about 3-6 times less effective against E. coli. The predicted complete inactivation time corresponding to the above D value was about 9 days and was within the limits of current cosmetic guidelines for bacteria of 'less than 0.1% survival by the 14th day.' PMID- 19291045 TI - Biosurfactants for cosmetic applications. AB - Synopsis Biosurfactants are biologically-produced, natural surface active agents and are synthesized by a range of microorganisms. These compounds include glycolipids, lipopeptides and phospholipids. The yields of biosurfactants in microbial fermentations may be as high as 150g l(-1) and may be increased by optimizing production conditions and by strain improvement. Biosurfactants display very low critical micelle concentration (CMC) values when compared with conventional surfactants and are stable at a wide range of temperatures, pH values and salinities. They show many promising characteristics for cosmetic applications. PMID- 19291046 TI - Ethylene formation from methionine as a method to evaluate oxygen free radical scavenging and metal inactivation by cosmetics. AB - Synopsis It has been proposed that oxygen free radicals are involved in skin aging. This paper describes a new method for the evaluation of oxygen free radical scavenging by cosmetic products. It is based on the measurement, by gas chromatography, of ethylene produced during the oxidation of methionine by the hydroxyl radical. OH. is produced by an iron catalyzed superoxide-driven Fenton reaction in which superoxide is obtained by photochemical oxygen reduction. The cosmetic is applied, together with methionine, riboflavine, NADH, FeCl(3) and EDTA, on a glass microfibre filter and submitted to UVA exposure through a quartz cell. Ethylene is then measured from aliquots of the atmosphere inside the cell. Catalase or Desferal completely inhibits ethylene production. SOD or high concentrations of hydroxyl radical scavengers (Mannitol, DMSO etc.) afford a partial protection. Thus the efficiency of O(2) (-)., H(2)O(2) and OH. scavengers and iron chelators can be measured. The main advantage of this test is that it is performed in conditions which simulate skin during UV exposure (e.g. air and UV exposed thin layer). Furthermore, as it is non-invasive, it can also be applied to human skin in vivo. PMID- 19291047 TI - [Not Available]. AB - Synopsis The normal dermal human fibroblastic cell (NDHF) was used to determine a cellular ageing pattern. Cells were cultured in monolayers until the 30th passage. First of all, the following cell growth characteristics were studied: growth rate, fluorimetric DNA determination, DNA repair after UV irradiation. Secondly, metabolism characteristics were examined: lysosomal enzymatic activity and type I and III collagen biosynthesis. Strains were obtained from 10,30,43 and 69-year-old donors to favour a comparison between in vitro and in vivo ageing. Cell growth ability is modified in vitro only for the oldest strain which shows a significant decrease in the cellular density at the 30th passage. The DNA rate and its repairing ability are not changed by in vitro ageing whatever the strain age. Lysosomal activity increases during in vitro ageing whereas the collagen I synthesis decreases. In vitro proliferating potentialities do not reflect in vivo ageing. On the other hand, in this study, metabolic potentialities evolve in the same way in vitro as in vivo and could be a good enough pattern to select anti ageing products. PMID- 19291048 TI - Influence of skin cleansing preparation acidity on skin surface properties. AB - Synopsis Two long-term trials were conducted each over eight weeks to compare the effect of the regular application of skin cleansing preparations of pH 5.5 and pH 8.5 and pH 5.5 and pH 7.0 respectively on the surface pH, roughness and transepidermal water loss (TEWL) of normal human forehead and forearm skin. Both trials were based on a cross-over design: five healthy volunteers started with a pH 5.5 preparation and switched to the other after four weeks, five additional volunteers used the preparations in the opposite order. While the skin surface pH was markedly lower in those individuals using the pH 5.5 preparation at each examination, as compared to those using the pH 8.5 or pH 7.0 preparation, no such difference could be established with respect to skin roughness and TEWL. Hence the skin irritancy of a cleansing preparation does not seem to be linked to its pH within the pH ranges tested. PMID- 19291049 TI - Skin lipids. AB - Synopsis Concepts regarding the structure and function of the stratum corneum (SC) have changed from that of a tough film of loosely adherent cells to that of a two-compartment system of protein-enriched cells embedded in intercellular lipids. The two-compartment arrangement enlarged the role of epidermal lipids from that of the plasticizing component of the SC to that of the epidermal barrier governing water-holding properties as well as take-up of water, the differences in permeability of topically applied lipophilic and hydrophilic agents as well as cohesion and desquamation of the SC. Barrier properties of SC are dependent largely on the intactness of the lipid lamellae that surround the corneocytes. The pliability of the SC depends on a correct balance of lipids, hygroscopic water-soluble substances and water. Mammalian differentiation involves characteristic changes in lipid composition consistent with the requirements for waterproofing. There is a progressive depletion of phospholipids and glycosphingolipids with enrichment in ceramides, cholesterol, free fatty acids, and small amounts of other polar (e.g. cholesterol sulphate) and non-polar species (e.g. hydrocarbons, cholesterol esters, triglycerides). The sphingolipids account for the most lipid by weight and are presumed to be of major importance for the water-retaining properties of the epidermal barrier. Decrease of sphingolipid content occurs in aged skin. Deficiency of essential fatty acids leads to enhanced transdermal water transport in addition to dryness and scaliness of the skin. A mixture of both saturated and unsaturated fatty acids produces the optimal barrier to water loss from the SC. The balance between solid crystal and liquid crystalline phases in epidermal lipids is determined by the degree of fatty acid unsaturation and the amount of water. PMID- 19291050 TI - Changes in glutathione content in human hair follicle keratinocytes as a function of age of donor: relation with glutathione dependent enzymes. AB - Synopsis Glutathione (GSH) plays an important role in cellular protection during ageing. The hair plucking technique is a non-invasive method for the direct biochemical study of keratinocytes. Hair was taken from the suboccipital area of 63 volunteers (men and women whose ages ranged from 13 to 103 years). The results show a diminution in the glutathione content as a function of age. We compare two groups of population: group A (less than 80) and group B (more than 80). A remarkable fact is observed: group B displays a weak dispersion of the values as compared to A. The glutathione content (nmol 10(-3) mg DNA) is 5.42 +/- 0.60 for A and 1.86 +/- 0.35 for B. A reduction of 88% was observed in glutathione reductase activity and of 78% in the activity of glutathione-S-transferase from group A to group B. The glutathione peroxidase activity remains relatively constant. The decrease in the GSH concentration and the constancy of the glutathione peroxidase suggest that the capacity of the cell to protect itself from peroxides remains unchanged but that the GSH concentration may become the limiting factor. PMID- 19291051 TI - [Not Available]. AB - Synopsis Antielastase activity of derivatives like 'propionylaminoacid'(C(3) prolin, C(3) hydroxyprolin, C(3) collagen) was examined for pancreatic elastase, and fibroblastic elastase production. Essential metabolic variations of normal dermal fibroblasts were evaluated: adhesion, proliferation capacity, total protein biosynthesis and collagen type I and type III production. Possible other factors such as cellular nutrients were examined by oxygen consumption evaluation. Propionylaminoacid derivatives have antielastase activities. Pancreatic elastase showed dose related inhibition (20% to 50% inhibition for concentration from 5 to 80 mg ml(-1). Moreover, fibroblastic elastase production was inhibited, cellular respiration was enhanced. A very good tolerance in vitro was observed for concentration 0-1 mg ml(-1) range: adhesion, proliferation capacity and collagen (type I and type III) production were not altered, and oxygen consumption was enhanced. PMID- 19291052 TI - Determination of alpha-bisabolol and d-panthenol in cosmetic products by gas chromatography. AB - Synopsis alpha-bisaboloi and d-panthenol are used in many cosmetic preparations, respectively, for their anti-inflammatory and regenerating properties. Their quantitative determination was an important element of their evaluation in these emulsions. Their concentration has been determined by gas chromatographic techniques, using a flame ionization detector. In both cases, the internal standards have been chosen for their compatibility with the analysis of extracted substances. Owing to the complexity of the cosmetic formulations, a preliminary extraction of alpha-bisabolol and d-panthenol was necessary. For the two substances the preparative separation was based on a liquid-liquid extraction. After dissolving the emulsion in methanol and diluting it with an aqueous buffer solution alpha-bisabolol was then extracted by ethyl acetate and d-panthenol by ethyl formate. PMID- 19291053 TI - Fine details on the surface of human hair. AB - Synopsis Some previously unreported fine variations in the form of normal hairs are described as they are observed in the scanning electron microscope (SEM). They all arise in the course of surface frictional wear and the chipping away of the hair surface scale edges and include remanent surface impressions of cuticle scale edges, 'false' scale edges, granular surface remnants and highly irregular (chevron) scale patterns. Mechanisms are proposed for the way in which each of these different fine features arise. The paper also contains deliberations on the conditions for operating the SEM consistent with obtaining the best information about the architecture of hair surfaces. The correctness of viewing orientation of scanning electron micrographs is also emphasized to avoid misinterpretation of features on the hair surface. PMID- 19291054 TI - Solution behaviour of new cationic surfactants derived from Guerbet alcohols and their use in hair conditioners. AB - Summary The solution behaviour of new cationic surfactants, synthesized by using long-chain Guerbet alcohols, in water was investigated by a polarizing microscopic technique, differential scanning calorimetry, and electric conductivity measurements. These surfactants show the gel-liquid crystalline phase transition to be lower than 0 degrees C and form lamellar liquid crystals even in cold water and at low concentrations of surfactants. The sorption of cationic surfactants from aqueous solutions onto hair was determined as a function of temperature. The effect of adsorbed cationic surfactants on the critical surface tension and kinetic frictional coefficients of hair surface have been investigated. These surface characteristics of hair treated with quaternary ammonium compounds derived from Guerbet alcohols were found to be significantly improved. These results can be explained by the high ability of sorption onto hair. Hair rinses and conditioners having excellent ease of combing or brushing for wet and dry hair can be formulated by the application of these cationic surfactants. PMID- 19291055 TI - The role of primary alkanolamines in cosmetic formulation. AB - Synopsis Alkaline products are commonly used in cosmetic preparations as neutralizing agents for acid-functional raw materials. Primary alkanolamines can provide positive attributes in such cases. The properties of nitroparaffin derived primary alkanolamines are compared here with the general requirements for cosmetic-type acid-neutralizing materials. Acid-functional copolymers, used as hair fixatives in both aerosol and non-aerosol products, are efficiently solubilized by neutralization with 2-amino-2-methyl-1-propanol (amino methylpropanol) in a variety of aqueous and non-aqueous systems. The effects of such neutralization on resultant film properties which influence product performance are discussed in detail. Upon neutralization, crosslinked acrylic acid polymers of varying molecular weights (carbomers) provide significant benefits when used as thickeners, gellants or emulsifiers in cosmetic products. New data are added to previous knowledge on the use of tris(hydroxymethyl)aminomethane (tromethamine) as a neutralizing agent for three resins, especially in gelled formulations. Alcohol compatibility, pH behaviour, and viscosity characteristics of such formulations are described. In addition, toxicological considerations of primary alkanolamines are addressed. Further to the above applications, nitroparaffin-derived primary alkanolamines can be effectively used as cosmetic emulsifiers (in the form of their fatty-acid soaps), dispersants or co-dispersants for particulate-containing products, and pH buffers/stabilizers. PMID- 19291056 TI - The use of Fourier transform infra-red spectroscopy in the study of the surface chemistry of hair fibres. AB - Synopsis Human hair keratin is susceptible to damage by both chemical and physical agencies. Oxidative damage can now be assessed by the use of Fourier transform infra-red (FTIR) spectroscopy. Analysis may be performed using the following techniques; diffuse reflectance, attenuated total reflectance and microspectrophotometry. This latter technique is made possible by the coupling of a research grade microscope to the optical bench of the FTIR spectrometer. The computing power necessary to operate the FTIR allows data to be mathematically manipulated. Derivative spectra obtained in this manner allow small peaks obscured by the close proximity of large peaks to be resolved. Analysis of single fibres treated with hydrogen peroxide or metabisulphite solutions showed the formation of cysteic acid and cysteine-S-sulphonate respectively. Second order derivative spectroscopy allowed a profile of the oxidative damage from the root to tip of naturally weathered hair to be constructed. PMID- 19291057 TI - Sodium lauryl sulphate/amphoteric surfactant interactions: selective diffusion through a collagen film. AB - Synopsis Diffusion tests of anionic, amphoteric and mixtures of amphoteric/anionic surfactants have been performed using a collagen film. A sensitive potentiometric method has been developed to determine the surfactant diffused avoiding the interferences observed in the usual analytical measurements due to the simultaneous presence of amphoteric/anionic surfactants. From the results obtained it is evident that the surfactant mixtures exhibit a synergistic behaviour as there is a mutual inhibition of the surfactant diffused through the collagen film. Empirical equations have been derived to describe the diffusion process of both the individual surfactants and mixtures of surfactants which allow the diffusion of the individual surfactants and the inhibition of diffusion in mixtures to be quantified. PMID- 19291059 TI - Simulated consumer exposure to dimethyl ether and propane/butane in hairsprays. AB - Synopsis The potential human exposures from use of dimethyl ether (DME) and 'liquefied petroleum gas'(LPG) arising from use in hairsprays have been assessed. DME and LPG concentrations were measured in the 'breathing zone' of an experimental manikin and an 'accompanying child' designed to simulate human use of hairsprays in a domestic situation and in the breathing zone of a 'stylist' and 'customer' under salon conditions. Results were expressed as the 10 min time weighted average in the air (TWA10) and as the peak concentration in the breathing zone of the 'user'. Following a 10s use of hairspray containing 50% DME or 26% LPG, TWA10 values for an adult user in a closed room (volume 21 m(3)) were on average 114 ppm and 73 ppm respectively. The child TWA10 values were 89 ppm (DME) and 80 ppm (LPG). Leaving the door open during spraying did not significantly alter these values. The peak concentrations measured in the user breathing zone were 1577 ppm of DME and 671 ppm of LPG. Simulated salon use of a hairspray gave a calculated value of 55 ppm DME and 88 ppm LPG for the stylist over an 8 h working period. PMID- 19291060 TI - Novel quaternary ammonium salts derived from triglycerides and their application in skin and hair products. AB - Synopsis An investigation has been carried out into the synthesis of quaternary ammonium salts of triglycerides. The aim of this work was to produce such compounds without breaking the triglyceride skeleton. The reaction of chloracetyl chloride with the triglyceride of ricinoleic acid which is the major constituent (88-90%) of castor oil gave an intermediate derivative with pendant chloro groups. Quaternization with stearyl dimethylamine gave a 98% yield of a quaternary ammonium salt containing three quaternary groups per molecule. This reaction sequence overcame the steric hindrance problems encountered in unsuccessful attempts to quaternize halogen groups attached directly to the main chain. Using the same procedure, a quaternary salt was prepared from castor oil. In skin care studies this quaternary was used at a low level in a moisturizing lotion. Subjective assessment of the ease of application of the test lotion versus control was significantly in favour of the test. Viscoelastic measurements of the skin were made using two techniques before and after application of the lotion. Both showed an advantage to the test lotion. No difference between the test and control lotions was found when sebum levels and transepidermal water loss measurements were measured. A C-12 quaternary synthesized from dimethyl dodecylamine exhibited antimicrobial activity. An aqueous dispersion of the C-18 quaternary when applied to bleached hair in order to improve combing, gave excellent results showing superiority to four other commercially available quaternary ammonium salts. PMID- 19291061 TI - Skin colour typology and suntanning pathways. AB - Synopsis The evaluation of sun-product efficacy, with laboratory solar simulators or in actual sun, implicates clinical and subjective assessment of the various skin responses in terms of wavelengths constitutive of solar light. These photobiological responses vary according to skin types and particularly to basic skin melanic content, i.e. with skin colour. Now, the instrumental measurement of live skin colour has become easier to perform, fast and reliable. Based on the standard CIE-L*a*b* colour system and correlated with the human eye, this technique was used to define the skin colour domain of the caucasian population, to propose a skin colour classification, and then to objectively follow, over a three week period, the dynamics and kinetics of tanning induced by UVB, UVA and UVB +/- A multi-exposures on the three skin categories. The specific directions in the three-dimensional L*a*b* colour space of the tanning components, i.e. erythema, immediate pigmentation and constitutional melanization, as well as the resulting tanning pathways, were analysed and defined in the three-dimensional colour space, using a vectorial method. The UVB, UVA and UVB +/- A tannings were differentiated by their intensity, their hue and especially their lasting capacity: UVA tanning clearly appeared more lasting than UVB. In addition, the UVA*UVB interaction on tanning intensity was not found to be significant. With the skin colour classification and the tanning models, this comprehensive study supplies a basic tool for the colorimetric interpretation of the skin phenomena involved, provided that this interpretation is always considered in the three dimensions of the colour space. It also suggests some useful practical applications for sun product formulation and evaluation. PMID- 19291062 TI - Optimization of emulsions. AB - Synopsis The optimization of three emulsions with predetermined components was performed by the aid of experimental design. Three different surfactants were compared: the salts of hexadecylhydrogenphosphate with diethanolamine, lysine and arginine respectively. Optimized microemulsions showed quite high stabilities towards centrifugation (30-60 min at 7000 rpm). The results showed that the counter-ion can affect the composition and stability of an emulsion. PMID- 19291063 TI - Skin surface profile technique and its applications. AB - Synopsis In this study the 'skin surface profile'(SSP) technique and its applications are described. With this technique the SSP is replicated by means of silicone impression materials. The SSP preparations were studied by a stereomicroscope or microscopic projector. Some applications of SSP technique are as follows: a. measurement of the stratum corneum hydration, b. study of the profile of primary crests and lines, c. measurement of the primary crests reservoir available for extension, and d. estimation of the true area of the skin surface in a body region. PMID- 19291064 TI - In vivo evaluation in man by two noninvasive methods of the stratum corneum barrier function after physical and chemical modifications. AB - Synopsis This study investigated the effect of different treatments on skin permeability, in vivo in man, by two noninvasive methods: transepidermal water loss (TEWL) determination measured with an evaporimeter, and Laser Doppler velocimetry (LDV) to measure the lag time before the vasodilatation induced by application of methyl and hexyl nicotinates. Different treatments were performed on forearms of volunteers: 1. removal of the stratum corneum by stripping, 2. occlusion and hydration of the stratum corneum, 3. application of three surfactants in aqueous solution (Tween 60, sodium dodecyl sulphate and cetyl trimethylammonium bromide). Increase in TEWL and decrease in lag time before the vasodilating effects of nicotinates measured with LDV after all treatments confirmed the modifications in skin permeability. The variations observed were more or less significant depending on the treatment, the investigative method, and the molecule used for the penetration study. With methyl (hydrophilic) nicotinate, the more TEWL increased, the more lag time decreased. A significant decrease in lag time after hexyl (lipophilic) nicotinate application was observed when there was a very high increase in TEWL. The lipophilic nature of the stratum corneum barrier was confirmed in vivo. Use of these two complementary methods, TEWL and LDV, allows the evaluation in vivo of the effect of different treatments on skin permeability in man. PMID- 19291065 TI - Identification des filtres UV dans les produits cosmetiques par CCM et HPLC. AB - Synopsis The method describes the identification of 22 UV filters authorized in Cosmetic products in Europe (dir. 76/768 EEC, annex VII, modified by dir. 89/174 EEC). (Table I). Twelve UV filters registered in part 2 of annex VII (dir. 83/574EEC) provisionally allowed in 1983 were abandoned by Industry. Consequently they were suppressed from the list of UV filters which cosmetic products may contain (dir. 89/174 EEC). Nevertheless the method describes also the identification of these 12 chemicals because they could interfere with the analysis of the 22 authorized UV filters in view of similarities in chemical structures (Table I (suite)). The screening of the UV filters in final products is based on thin layer chromatography (TLC) and confirmed by high pressure liquid chromatography (HPLC/UV). TLC combines four different systems and several detection reagents: absorption chromatography on silica gel with development in a neutral solvent, an acidic solvent, a basic solvent, reversed phase chromatography on RP 18 bonded silica with development in a methanol/water mixture. The results of these screening tests are confirmed by reversed phase HPLC/UV in an isocratic elution system. The acid components and the salts which are eluted before or with the solvent peak are separated in a gradient elution system at pH 3.5. Several UV filters available on the market of cosmetic ingredients are impure chemicals or mixtures. The identification of their principal components by nuclear magnetic resonance spectrometry (NMR) and by gas chromatography/mass spectrometry (GC/MS) will be the subject of future publications. PMID- 19291067 TI - Pigment loss in response to the environment: a new role for the WD/bHLH/MYB anthocyanin regulatory complex. PMID- 19291068 TI - Chlorophyll and folate: intimate link revealed by drug treatment. PMID- 19291069 TI - Plant defense: a pre-adaptation for pollinator shifts. PMID- 19291070 TI - Individuals, populations, communities and function: the growing field of ectomycorrhizal ecology. PMID- 19291071 TI - Mycorrhizas in tropical forests: a neglected research imperative. PMID- 19291074 TI - The GO system prevents ROS-induced mutagenesis and killing in Pseudomonas aeruginosa. AB - Inactivation of the Pseudomonas aeruginosa mutM, mutY, or mutT gene conferred a 2.4-, 17.2-, or 38.1-fold increase in spontaneous mutation frequency, respectively. Importantly, the mutY and mutT strains each displayed a robust H(2)O(2)-induced mutation frequency. In addition, the mutM, mutY, and mutT mutations severely sensitized P. aeruginosa to killing by H(2)O(2), suggesting that these gene products act to repair one or more cytotoxic lesions in P. aeruginosa. Nucleotide sequence analysis of a fragment of the rpoB gene from rifampicin resistant mutM-, mutY-, and, mutT-deficient strains was consistent with this conclusion. These findings are discussed in terms of possible roles for mutM, mutY, and mutT in contributing to survival and mutagenesis of P. aeruginosa colonizing the airways of cystic fibrosis patients. PMID- 19291075 TI - In-patient suicide--a 13-year assessment. AB - OBJECTIVE: To describe the suicide rates of psychiatric in-patients in the canton of Zurich for the period 1992-2004, and to determine putative risk factors. METHOD: The data were derived from the psychiatric case register of the canton Zurich. The analyses were based on person-years calculations and standardised mortality ratios. Additional information was assessed via case records. RESULTS: During the 13-year period the standardised mortality ratio was 48.9. The risk of suicide was particularly high in patients with personality and affective disorders. Most suicides occurred during regular leave periods, despite the fact that clinical assessment had indicated there was no suicide risk for the patient. CONCLUSION: The suicide risk of in-patients is distinctly higher than in the general population. A better assessment of suicide risk before regular leave periods could lead to a decrease of suicides in in-patient settings, as well as a more rigorous treatment of borderline cases, and of affective and psychotic symptoms. PMID- 19291077 TI - Atypical depression: retrospective self-reporting of treatment effectiveness. AB - OBJECTIVE: Earlier studies demonstrated that those with atypical depression show a differentially superior response to monoamine oxidase inhibitor (MAOI) antidepressants. This study compares ratings of effectiveness for a range of treatments, amongst depressed subjects with and without atypical features. METHOD: In an on-line survey, individuals experiencing likely clinical depression rated symptoms experienced when depressed, including 'atypical features' and the effectiveness of previous treatments. Mean treatment effectiveness ratings were compared amongst those with 'atypical depression' (n = 338) and 'non-atypical depression' (n = 377). RESULTS: There were few significant differences between the 'atypical depression' and 'non-atypical depression' groups in effectiveness ratings for drug treatments, and none for psychological treatments. The 'atypical depression' group had significantly lower mean effectiveness ratings for some selective serotonin reuptake inhibitor antidepressants. Few respondents had trialed MAOIs. CONCLUSION: While MAOIs are rarely prescribed, a range of non-MAOI drug and psychological treatments are of some perceived benefit for depressed patients with atypical features. PMID- 19291078 TI - Burning mouth syndrome and electroconvulsive therapy. PMID- 19291079 TI - Violence against people with severe mental illness in Europe. PMID- 19291080 TI - Psychotic and behavioural symptoms in a population-based sample of the very elderly subjects. AB - OBJECTIVE: The aim was to elucidate the relationship between psychotic and behavioural symptoms in the elderly. METHOD: A representative sample of 85 year old subjects living in Gothenburg, Sweden (n = 451) was assessed with neuropsychiatric examinations, key informant interviews and record reviews. RESULTS: Fourteen percent of these very elderly subjects had paranoid symptoms with concomitant anxious agitation and/or irritability/anger. Hallucinations and paranoid symptoms were both associated with a pattern of behavioural symptoms including both anxious agitation and irritability/anger simultaneously in both demented [hallucinations, Odds ratio (OR) 2.8, Confidence interval (CI) 1.2-6.7, paranoid symptoms OR 5.6 CI 2.2-14.2] and non-demented (hallucinations OR 3.2 CI 1.2-8.3, paranoid symptoms OR 4.8 CI 2.0-11.8). CONCLUSION: Psychotic symptoms are associated with behavioural symptoms regardless of dementia status. Since these symptoms lead to decreased ability to function in daily life and increased caregiver burden, it is important for health professionals to identify and treat these symptoms also in non-demented. PMID- 19291081 TI - Obsessive-compulsive disorder: a disorder of pessimal (non-functional) motor behavior. AB - OBJECTIVE: To determine whether in addition to repetitiveness, the motor rituals of patients with obsessive-compulsive disorder (OCD) involve reduced functionality due to numerous and measurable acts that are irrelevant and unnecessary for task completion. METHOD: Comparing motor rituals of OCD patients with behavior of non-patient control individuals who were instructed to perform the same motor task. RESULTS: Obsessive-compulsive disorder behavior comprises abundant acts that were not performed by the controls. These acts seem unnecessary or even irrelevant for the task that the patients were performing, and therefore are termed 'non-functional'. Non-functional acts comprise some 60% of OCD motor behavior. Moreover, OCD behavior consists of short chains of functional acts bounded by long chains of non-functional acts. CONCLUSION: The abundance of irrelevant or unnecessary acts in OCD motor rituals represents reduced functionality in terms of task completion, typifying OCD rituals as pessimal behavior (antonym of optimal behavior). PMID- 19291083 TI - Effects of a communication course for clinicians on parents' perception of care- a randomized controlled trial. AB - INTRODUCTION: In paediatric care, it is a great challenge to make sure that all parties feel involved and heard, they all have had the opportunity to take part in the decisions and all the participants experienced that their expectations to the consultation are fulfilled. Previous research in communication skills training has primarily focused on the behavioural changes of clinicians, and only few studies have investigated the effect from the perspective of the user. METHOD: A randomized controlled trial including medical doctors and nurses from paediatric outpatients' clinics was carried out. The intervention group completed a 5-day communication course, whereas the control group had no intervention. The intervention was evaluated using questionnaires measuring parents' perception of the communication and their satisfaction. The questionnaires were filled out by parents to children consulting a clinician in the outpatient's clinic. RESULTS: Before the intervention, 75% (314/419) answered the questionnaire and 65% (271/419) were included in the precourse analysis. After the intervention, 68% (946/1395) answered the questionnaire and 55% (764/1395) were included in the postcourse analysis. There were no significant differences between the satisfaction of parents visiting clinicians from the intervention group and those visiting clinicians from the control group; however, the proportion of parents who had a positive perception of the communication was up to 9.8% higher in the intervention group compared with the control group. For example: 'the clinician told my child what he/she could do in order to feel better'. DISCUSSION: Although no statistically significant differences were found, the study indicates that parents who had visited a clinician from the intervention group have experienced the communication as more positive. PMID- 19291084 TI - Conditions for successfully implementing resident-oriented care in nursing homes. AB - This study reports an investigation of the conditions for a successful introduction of a resident-oriented care model on six somatic and psychogeriatric intervention wards in three Dutch nursing homes. This study aims to answer the following research question: 'What are the conditions for successfully implementing resident-oriented care?' To answer the research question, the organisational change process was monitored by using the '7-S' model of Peters and Waterman as a diagnostic framework. Based on this model, the following change characteristics were studied: structure, strategy, systems, staff, skills, style and shared values. Our study involved a one group pretest/post-test design. To measure the conditions for change, we operationalised the factors of the 7-S model serving as a diagnostic framework and studied their presence and nature on the intervention wards. For this purpose qualitative interviews were held with the change agents of the nursing homes and the wards' supervisors. To determine the degree of 'success' of the implementation, we measured the extent to which resident-oriented care was implemented. For this purpose a quantitative questionnaire was filled in by the nurses of the intervention wards. By relating the extent to which resident-oriented care was implemented to the differences in change conditions, we were able to distinguish the 'most' from the 'least' successful intervention ward and so, pointing out the conditions contributing to a successful implementation of resident-oriented care. The results showed that, in contrast to the least successful intervention ward, the most successful intervention ward was characterised by success conditions related to the 7-S model factors strategy, systems, staff and skills. The factor structure did not contribute to the success of the implementation. Success conditions appeared to be related to the ward level and not to the organisational or project level. Especially the supervisors' role appeared to be crucial for a successful implementation. PMID- 19291086 TI - Should edentulous patients be constrained to removable complete dentures? The use of dental implants to improve the quality of life for edentulous patients. AB - BACKGROUND: Nowadays, there is some speculation among dental educators that the need for complete dentures will significantly decrease in the future and that training in their provision should be removed from the dental curriculum. OBJECTIVE: To sensitise the reader to the functional shortcomings of complete denture therapy in the edentulous patient and present restorative options including implants to improve edentulous quality of life in these patients. METHODS: Information retrieval followed a systematic approach using PubMed. English articles published from 1964 to 2008, in which the masticatory performance of patients with implant-supported dentures was assessed by objective methods and compared with performance with conventional dentures, were included. RESULTS: National epidemiological survey data suggested that the adult population in need of one or two complete dentures will increase from 35.4 million adults in 2000 to 37.9 million adults in 2020. Clinical studies have showed that the ratings of general satisfaction were significantly better in the patients treated with implant overdentures post-delivery compared with the complete denture users. In addition, the implant group gave significantly higher ratings on comfort, stability and ability to chew. Furthermore, patients who received mandibular implant overdentures had significantly fewer oral health-related quality of life problems than did the conventional group. CONCLUSION: Implant-supported dentures including either complete overdentures or a hybrid prosthesis significantly improve the quality of life for edentulous patients compared with conventional removable complete dentures. Therefore, the contemporary dental practitioner should consider other options as well as conventional removable complete dentures to restore edentulous patients. PMID- 19291087 TI - The role of nitric oxide- and prostacyclin-independent vasodilatation in the human cutaneous microcirculation: effect of cytochrome P450 2C9 inhibition. AB - The component of the flow- or agonist-dependent vasodilatation, insensitive to inhibitors of nitric oxide (NO) synthases (NOS) or cyclooxygenases (COX), is suggested to reflect the production of an endothelium-dependent hyperpolarizing factor (EDHF). The identity of EDHF in humans remains controversial; in coronary arterioles, it appears to be a cytochrome P450 (CYP) 2C9-derived metabolite, whereas there are no data for human skin microcirculation. Therefore, the aim of our study was to investigate the role of the NO- and prostacyclin (PGI(2)) independent mechanism, particularly the potential involvement of CYP 2C9, in skin microcirculation. We measured skin blood flow on the volar aspect of the forearm in 12 healthy subjects by laser-Doppler fluxmetry (LDF). The inhibitors of NOS, N(omega)-monomethyl-L-arginine (L-NMMA), and cyclooxygenase (COX), diclofenac, as well as sulfaphenazole, the specific CYP 2C9 inhibitor, and saline as control, were administered to the measurement sites by an intradermal microinjection in different combinations. Afterwards, baseline LDF was assessed and iontophoresis of acetycholine (ACh) applied. Combined NOS and COX inhibition had no effect on baseline LDF, whereas it significantly reduced the ACh-induced increase in LDF (t test, P<0.05). Sulfaphenazole did not affect baseline LDF either in the control site or in the L-NMMA- and diclofenac-pretreated site. In addition, sulfaphenazole did not attenuate the ACh-induced vasodilatation in either site. We conclude that a NO- and PGI(2)-independent vasodilator mechanism, potentially attributable to EDHF, contributes substantialy to the ACh-induced vasodilatation in human skin microcirculation and that it is probably not a CYP 2C9-derived metabolite. PMID- 19291088 TI - Characterization of vaginal lactobacilli coaggregation ability with Escherichia coli. AB - The coaggregation abilities of probiotic strains might enable it to form a barrier that prevents colonization by pathogenic bacteria. In the present study, the characterization of the coaggregation ability of 19 vaginal lactobacilli was studied. Coaggregation ability of all lactobacilli with Escherichia coli ATCC 11229 was positive. Only the highest coaggregation percentage of Lactobacillus acidophilus S1 was obtained with E. coli ATCC 11229 under both aerobic (71%) and anaerobic conditions (62%). The coaggregation abilities of strains occurred higher at acidic pH than at basic pH values. Moreover, the coaggregation abilities of tested strains against E. coli decreased after heat treatment (70 or 85 degrees C). Also, the relationship between hydrophobicity and coaggregation of strains was found to be significant. The effect of sonication, some enzymes (lipase and pepsin) and sodium periodate on coaggregation ability of L. acidophilus S1, which is one of the highest potentials on coaggregation ability, was investigated. Sodium periodate did not have a significant effect on coaggregation ability of L. acidophilus S1. The sonicated cell showed lower coaggregation than the control, the supernatant fluid of this sonicated cells showed similar coaggregation ability to the control. Coaggregation abilities of bacteriotherapeutic lactobacilli with pathogenic bacteria can be used for preliminary screening in order to identify potentially probiotic bacteria suitable for human use against urogenital tract infections. PMID- 19291085 TI - The impact of dose escalation and resistance modulation in older patients with acute myeloid leukaemia and high risk myelodysplastic syndrome: the results of the LRF AML14 trial. AB - The acute myeloid leukaemia (AML)14 trial addressed four therapeutic questions in patients predominantly aged over 60 years with AML and High Risk Myelodysplastic Syndrome: (i) Daunorubicin 50 mg/m(2) vs. 35 mg/m(2); (ii) Cytarabine 200 mg/m(2) vs. 400 mg/m(2) in two courses of DA induction; (iii) for part of the trial, patients allocated Daunorubicin 35 mg/m(2) were also randomized to receive, or not, the multidrug resistance modulator PSC-833 in a 1:1:1 randomization; and (iv) a total of three versus four courses of treatment. A total of 1273 patients were recruited. The response rate was 62% (complete remission 54%, complete remission without platelet/neutrophil recovery 8%); 5-year survival was 12%. No benefits were observed in either dose escalation randomization, or from a fourth course of treatment. There was a trend for inferior response in the PSC-833 arm due to deaths in induction. Multivariable analysis identified cytogenetics, presenting white blood count, age and secondary disease as the main predictors of outcome. Although patients with high Pgp expression and function had worse response and survival, this was not an independent prognostic factor, and was not modified by PSC-833. In conclusion, these four interventions have not improved outcomes in older patients. New agents need to be explored and novel trial designs are required to maximise prospects of achieving timely progress. PMID- 19291089 TI - Effects of Clinacanthus siamensis leaf extract on influenza virus infection. AB - Ethanolic extracts of 20 medicinal plants were screened for influenza virus NA inhibition and in vitro antiviral activities using MDCK cells in an MTT assay. The vaccine proteins of influenza virus A/New Caledonia/20/99 (H1N1), mouse adapted influenza virus A/Guizhou/54/89 (A/G)(H3N2) and mouse-adapted influenza virus B/Ibaraki/2/85 (B/I) were used in the NA inhibition assay, and mouse adapted influenza viruses A/PR/8/34 (H1N1), A/G and B/I were used in the in vitro antiviral assay. The results of the in vitro antiviral assay indicated that the A/G virus was the most susceptible and an extract of the leaf of CS possessed the highest in vitro anti-A/G virus activity (41.98%). Therefore, the A/G virus and the CS extract were selected for studying in vivo anti-influenza virus activity. BALB/c mice were treated with CS extract (100 mg/kg per day, 5 times) orally from 4 hr before to 4 days after infection. CS extract elicited significant production of anti-influenza virus IgG(1) antibody in BAW and increased mouse weight compared to oseltamivir (0.1 mg/kg per day) on day 19 or water on days 17-19 of infection. Moreover, CS extract produced a higher anti-influenza virus IgA antibody level in BAW compared to oseltamivir, and a tendency towards an increase in anti-influenza virus IgA compared to water was shown. The results suggest that CS extract has a protective effect against influenza virus infection. PMID- 19291090 TI - Neutralizing antibody against severe acute respiratory syndrome (SARS) coronavirus spike is highly effective for the protection of mice in the murine SARS model. AB - We evaluated the efficacy of three SARS vaccine candidates in a murine SARS model utilizing low-virulence Pp and SARS-CoV coinfection. Vaccinated mice were protected from severe respiratory disease in parallel with a low virus titer in the lungs and a high neutralizing antibody titer in the plasma. Importantly, the administration of spike protein-specific neutralizing monoclonal antibody protected mice from the disease, indicating that the neutralization is sufficient for protection. Moreover, a high level of IL-6 and MCP-1 production, but not other 18 cytokines tested, on days 2 and 3 after SARS-CoV infection was closely linked to the virus replication and disease severity, suggesting the importance of these cytokines in the lung pathogenicity of SARS-CoV infection. PMID- 19291091 TI - Differences in clinical manifestations of influenza-associated encephalopathy by age. AB - Data from patients in Japan was analyzed to examine the age distribution and differences by age in the clinical manifestations of influenza-associated encephalopathy. Between 1998 and 2002, 472 cases of influenza-associated encephalopathy in patients aged 15 years or younger were reported to the Collaborative Study Group on Influenza-Associated Encephalopathy. These cases were divided into two groups by age: 0-5 and 6-15 years. The differences between the groups were estimated based on the data for those aged 0-5 years, and the odds ratios and 95% confidence intervals calculated. Distribution was inversely correlated with age, with a peak at 1-2 years old. In comparison with patients aged 0-5, those aged 6-15 years had a significantly greater incidence of type B infection, lower frequency of convulsions, higher frequency of loss of consciousness and altered consciousness as the initial neurological symptom, lower serum transaminase levels, lower frequency of low-density area for brain CT upon admission, and lower incidence of sequelae. Our analysis indicates that the clinical course, laboratory data, and brain imaging findings of influenza associated encephalopathy exhibits patterns that vary with age. PMID- 19291092 TI - The context of tetanus toxoid application influences the outcome of antigen specific and self-directed humoral immune response. AB - Results are presented concerning our attempts to create a suitable model system for studying the connection between microbial antigen (micAg), autoimmunity and autoimmune disease on the basis of hyper-immunization and application of micAg in different contexts. Our research was focused on tetanus toxoid (TTd) as a model micAg. Non-pretreated and complete Freund's adjuvant pretreated BALB/c mice were immunized with high doses of TTd mixed with glycerol or aluminum hydroxide as adjuvants. The main aims of the experiments were to evaluate the properties of induced humoral immune responses, evaluate the pathological potential of induced immune responses and determine possible correlations between the properties of a humoral immune response and its pathological potential. The production of TTd specific and self-reactive beta(2)-glycoprotein I (beta(2)-GP I)-specific antibodies (Abs) was detected in all groups but with specific, context-related properties. Analysis of pregnancy-related pathology (anti-beta(2)-GP I Abs associated) showed differences in the pathological potential of the induced immune response. It was demonstrated that severity of pathology is positively correlated to the abundance of IgG that recognizes beta(2)-GP I adsorbed onto phosphatidylserine, and to IgG affinity. Furthermore, it was demonstrated that molecular mimicry, which results in generation of anti-beta(2)-GP I Abs upon TTd immunization, is necessary but not sufficient for the development of pregnancy related pathology. PMID- 19291093 TI - Presence of a novel Ehrlichia sp. in Ixodes granulatus found in Okinawa, Japan. AB - Ehrlichia-specific DNA fragments of Ehrlichia omp-1 and groEL genes were found in two I. granulatus ticks which had been collected from wild small mammals in a subtropical zone in Japan. The DNA sequences of groEL and 16SrDNA of the suspected Ehrlichia were clustered into a group of E. chaffeensis, E. muris, and Ehrlichia sp. HF565 found in I. ovatus, but were distinctly different. Therefore the Ehrlichia strain was designated as a novel Ehrlichia sp. 360. The Ehrlichia sp. 360 was detected in I. granulatus but not in any other ticks. This suggests that I. granulatus is a probable vector of Ehrlichia sp. 360 in Japan. PMID- 19291094 TI - Prevalence of antimicrobial resistance among serotypes of Campylobacter jejuni isolates from cattle and poultry in Japan. AB - Penner serotypes of C. jejuni in a total of 601 isolates from apparently healthy cattle, layer and broiler chickens in Japan were examined between 2001 and 2006. Predominant serotypes were B (O: 2, 19.1%), D (O: 4, 13.5%), Y (O: 37, 7.3%) and G (O: 8, 5.8%), whereas the remaining serotypes made up less than 5% of the total isolates. The frequency of ampicillin resistance in serotype G (65.6%) was significantly higher than in serotypes D (12.5%), B (11.2%), and Y (0%). Our results suggest that serotype is one factor contributing to the prevalence of ampicillin resistance in C. jejuni isolates. PMID- 19291095 TI - New ITS genotype of Cryptococcus gattii isolated from an AIDS patient in Brazil. AB - Based on combinations of nine variable nucleotides at nine different base positions in the ITS1-5.8S-ITS2 region, Cryptococcus gattii strains were classified into six genotypes. A new genotype of C. gattii, designated as ITS type 8, was isolated from an AIDS patient in Brazil. The ITS type 8 strain is closely related to the ITS type 4 strain, which has been frequently isolated in Brazil and the USA, but which shows ITS-signatured nucleotide difference at each nucleotide position. The ITS type 8 strain is also differentiated from all heretofore reported ITS types of C. gattii strains in the RAPD band patterns and IGS sequence information. PMID- 19291096 TI - Comparison of different methods for detecting methicillin resistance in MRSA isolates belonging to international lineages commonly isolated in the American continent. AB - The aim of the present paper was to compare different methods for detecting methicillin resistance in Staphylococcus aureus. Among the isolates analyzed, 52 belonged to MRSA international lineages commonly detected in the American continent and 14 to sporadic MRSA clones. Both 30 microg-cefoxitin disk and PBP2a had 100% sensibility/specificity when the low-level heterogeneous isolates were tested and, thus, are highly recommended. PMID- 19291097 TI - Activation of mitogen-activated protein kinases is involved in the induction of interferon beta gene in macrophages infected with Orientia tsutsugamushi. AB - We investigated the role of MAPK in IFN-beta gene expression in macrophages after infection with Orientia tsutsugamushi. ERK1/2 became phosphorylated in Orientia stimulated macrophages. Selective inhibition of ERK1/2 and p38 MAPK could all significantly reduce Orientia-stimulated IFN-beta mRNA expression. Orientia inactivation by heat abolished IFN-beta mRNA induction only, whereas cytochalasin D treatment completely blocked both IFN-beta and chemokine expression, suggesting requirement of cellular internalization by viable bacteria for IFN-beta gene induction. In conclusion, our data indicate that MAPK pathways are required to induce maximal IFN-beta gene expression in macrophages during Orientia infection. PMID- 19291098 TI - Evaluating the ability of tree-based methods and logistic regression for the detection of SNP-SNP interaction. AB - Most common human diseases are likely to have complex etiologies. Methods of analysis that allow for the phenomenon of epistasis are of growing interest in the genetic dissection of complex diseases. By allowing for epistatic interactions between potential disease loci, we may succeed in identifying genetic variants that might otherwise have remained undetected. Here we aimed to analyze the ability of logistic regression (LR) and two tree-based supervised learning methods, classification and regression trees (CART) and random forest (RF), to detect epistasis. Multifactor-dimensionality reduction (MDR) was also used for comparison. Our approach involves first the simulation of datasets of autosomal biallelic unphased and unlinked single nucleotide polymorphisms (SNPs), each containing a two-loci interaction (causal SNPs) and 98 'noise' SNPs. We modelled interactions under different scenarios of sample size, missing data, minor allele frequencies (MAF) and several penetrance models: three involving both (indistinguishable) marginal effects and interaction, and two simulating pure interaction effects. In total, we have simulated 99 different scenarios. Although CART, RF, and LR yield similar results in terms of detection of true association, CART and RF perform better than LR with respect to classification error. MAF, penetrance model, and sample size are greater determining factors than percentage of missing data in the ability of the different techniques to detect true association. In pure interaction models, only RF detects association. In conclusion, tree-based methods and LR are important statistical tools for the detection of unknown interactions among true risk-associated SNPs with marginal effects and in the presence of a significant number of noise SNPs. In pure interaction models, RF performs reasonably well in the presence of large sample sizes and low percentages of missing data. However, when the study design is suboptimal (unfavourable to detect interaction in terms of e.g. sample size and MAF) there is a high chance of detecting false, spurious associations. PMID- 19291099 TI - Discovery and binding studies on a series of novel Pin1 ligands. AB - Pin1 plays a key role in various biological cellular processes via the recognition of phosphorylated Ser/Thr-Proline motifs. Moreover, high expression levels of Pin1 are correlated to tumorgenesis in some cancer types. Here, we identify a novel series of small molecular weight compounds with a core structure mimicking the phoshorylated serine. The binding affinity and binding mode of the compounds for Pin1 are analyzed via NMR spectroscopy and computational studies. The reported chemical probes and relative binding data to Pin1 represent valuable stepping stones for the validation of Pin1 as target for drug discovery and for eventually the development of possible lead compounds. PMID- 19291100 TI - Rational design of multitargeted tyrosine kinase inhibitors: a novel approach. AB - The non-receptor Src tyrosine kinase is known to cooperate with the epidermal growth factor receptor in a mechanism leading to invasion and metastasis of solid tumours. With the purpose of developing agents targeted to both epidermal growth factor receptor and Src or related kinases, we embarked on the design of chimeric molecules termed combi-molecules capable of blocking both Src and epidermal growth factor receptor. To this end, we have chosen to design molecules containing a quinazoline moiety (directed at epidermal growth factor receptor) and a 7-phenyl-pyrazolopyrimidine (directed at Src). Molecular modelling showed that the optimal position to attach the linker was the 6-position of the quinazoline and the 9-position of the pyrazolopyrimidine. This has led to the synthesis of SB162, SB166 and SB163. SB163 containing the longest linker was the only molecule capable of inducing a dose-dependent inhibition of both Src and epidermal growth factor receptor. SB163 also induced a dose inhibition of Abl and PDGFR. PMID- 19291101 TI - Design of conformationally constrained azole antifungals: efficient synthesis and antifungal activity of trans-3-imidazolylflavanones. AB - An efficient protocol has been developed for the preparation of a series of trans 3-imidazolylflavanones and (Z)-trans-3-imidazolylflavanone oximes, as potential antifungal agents, by the reaction of 2-imidazolyl-2'-hydroxyacetophenone with different benzaldehyde derivatives and subsequence oximation reaction. The stereochemical and conformational aspects of compounds were also assigned by (1)H NMR spectroscopy. The aryl group present at C-2 and the imidazole ring present at C-3 were assigned to occupy equatorial position in trans-3-imidazolylflavanones 3 and axial position in (Z)-trans-3-imidazolylflavanone oximes 4. Most of the 3 imidazolylflavanone derivatives containing a substituent on phenyl ring showed significant antifungal activity and modification of the 4-oxo group to oxime or substituent on phenyl ring produced changes in the antifungal activity profile of these compounds. Generally, 3'-chloro- analog of 3-imidazolylflavanone 3c exhibited better profile of antifungal activity against strains of fungi tested, superior than reference drug fluconazole. PMID- 19291102 TI - Dye labelling of nucleosides. AB - Dye-labelled nucleosides were obtained in 30-79% (average 45%) yields by treating N-(4-arylazobenzoyl)-1H-benzotriazoles 3a-b with appropriate nucleosides. Similarly, 3a-b afforded dye-labelled threoninol conjugates in 55-89% (average 67%) yields. All novel products were characterized by NMR and elemental analysis. PMID- 19291103 TI - Synthesis, biological evaluation, mechanism of action and quantitative structure activity relationship studies of chalcones as antibacterial agents. AB - Forty-eight chalcone analogs were synthesized and their in vitro antibacterial activity against Staphylococcus aureus NCIM 5021, Bacillus subtilis NCIM 2718, Phaseolus vulgaris NCIM 2813, Escherichia coli NCIM 2931, Salmonella typhi 2501 and Enterobacter aerogenes NCIM 5139 were evaluated by microdilution broth assay. Quantitative structure-activity relationships were developed for all the cases (r(2) = 0.68-0.79; r(2)(adj) = 0.58-0.78; q(2) = 0.51-0.68; F = 13.02-61.51). Size, polarizability, electron-donating/withdrawing and hydrophilic nature of the molecule determine the activity against these Gram-positive and Gram-negative bacteria. Staphylococcus aureus was the most and S. typhi was the least hydrophobic of these organisms. These chalcones act better against more hydrophobic organisms. The more active chalcones have log P between 1.5 and 3. Compound 24, one of the most active compounds, was found to act by damaging the cell wall of S. aureus. Slimicidal activity of five of the most active compounds (24, 31, 32, 34 and 37) was found to be in the range of 48-60% against S. aureus and 40-54% against E. coli. A correlation was observed among the hydrophobicity of the compounds, hydrophobicity of the bacterial cell surface and the antibacterial activity of the compound. PMID- 19291104 TI - Three-dimensional quantitative structure-activity relationship studies on c-Src inhibitors based on different docking methods. AB - c-Src kinase play an important role in cell growth and differentiation and its inhibitors can be useful for the treatment of various diseases, including cancer, osteoporosis, and metastatic bone disease. Three dimensional quantitative structure-activity relationship (3D-QSAR) studies were carried out on quinazolin derivatives inhibiting c-Src kinase. Molecular field analysis (MFA) models with four different alignment techniques, namely, GLIDE, GOLD, LIGANDFIT and Least squares based methods were developed. glide based MFA model showed better results (Leave one out cross validation correlation coefficient r(2)(cv) = 0.923 and non cross validation correlation coefficient r(2)= 0.958) when compared with other models. These results help us to understand the nature of descriptors required for activity of these compounds and thereby provide guidelines to design novel and potent c-Src kinase inhibitors. PMID- 19291105 TI - QSAR studies of CYP2D6 inhibitor aryloxypropanolamines using 2D and 3D descriptors. AB - Thirty six aryloxypropanolamine compounds with cytochrome 2D6 (CYP2D6) inhibitory activity were subjected to molecular shape analysis and molecular field analysis studies to explore the required molecular shape features as well as information on putative interactions with the active site of the enzyme. In addition to the 3D QSAR models, impact of two-dimensional (2D; thermodynamic, structural and topological including E-state parameters) descriptors towards the inhibitory activity was also studied. The whole data set was divided into training (n = 26) and test (n = 10) sets by K-means clustering technique. The chemometric tools used for molecular shape analysis and molecular field analysis were GFA and G/PLS techniques respectively. The G/PLS model derived in molecular field analysis using maximum common subgroup alignment was found to be the best model based on highest external (R(2)(pred) = 0.912) predictive power with lowest RMSEP value (0.363). The molecular field analysis-derived models suggest the requirement of U shape conformation for optimum interactions as well as importance of different substituents on the aryloxypropanol fragment like indolylalkyl substituent. The molecular shape analysis models indicate the importance of distribution of positive and negative charges on the surface of the molecules. The QSAR models with 2D descriptors reveal the importance of bulk, branching and presence of different fragments. PMID- 19291106 TI - Modelling of cytotoxicity data (CC50) of anti-HIV 1-[5-chlorophenyl) sulfonyl]-1H pyrrole derivatives using calculated molecular descriptors and Levenberg Marquardt artificial neural network. AB - A nonlinear quantitative structure anti-HIV activity relationship study was presented for modelling and predicting pyrryl aryl sulfones cytotoxicity data. Levenberg-Marquardt artificial neural network was used to link molecular structures and cytotoxicity data. A data set consisting of 27 derivatives of 1-[5 chlorophenyl) sulfonyl]-1H-pyrrole was used in this study. Among a large number of calculated descriptors, only eight significant molecular descriptors were obtained by stepwise regression, as the most feasible descriptors, and then they were used as inputs for neural network. The data set was randomly divided into 20 training and 7 validation sets and the neural network architecture and its parameters were optimized. The prediction ability of the model was evaluated using the validation data set, leave-one-out cross-validation and response randomization method. The mean square errors and mean absolute errors for the validation data set were 0.0067 and 0.066, respectively, and for the leave-one out method, they were 0.013 and 0.087, respectively. The results obtained showed the excellent prediction ability and stability of the proposed model in the prediction of cytotoxicity data of the corresponding anti-HIV analogues. PMID- 19291107 TI - 3-(3-ethylphenyl)-2-substituted hydrazino-3H-quinazolin-4-one derivatives: new class of analgesic and anti-inflammatory agents. AB - A new series of 3-(3-ethylphenyl)-2-substituted hydrazino-3H-quinazolin-4-ones were synthesized by reacting the amino group of 2-hydrazino-3-(3-ethylphenyl)-3H quinazolin-4-one with a variety of aldehydes and ketones. The title compounds were investigated for analgesic, anti-inflammatory and ulcerogenic index behavior. The compound 2-(N'-3-pentylidene-hydrazino)-3-(3-ethylphenyl)-3H quinazolin-4-one (AS2) emerged as the most active compound in exhibiting analgesic activity and the compound 2-(N'-2-pentylidene-hydrazino)-3-(3 ethylphenyl)-3H-quinazolin-4-one (AS3) emerged as the most active compound in exhibiting anti-inflammatory activity; and these compounds are moderately potent when compared with the reference standard diclofenac sodium. Interestingly, the test compounds showed only mild ulcerogenic potential when compared with aspirin. PMID- 19291109 TI - Diabetes and landmine-related amputations: a call to arms to save limbs. PMID- 19291111 TI - Extracorporeal shock wave therapy suppresses the early proinflammatory immune response to a severe cutaneous burn injury. AB - Following severe burn injury, persistent inflammation perpetuated by surface eschar, bacterial colonisation and neutrophil proteolytic activity can impede normal healing and result in further tissue damage. Extracorporeal shock wave treatment (ESWT) has been shown in the clinical setting to promote the healing of burn and difficult-to-heal wounds; however, the mechanism is unclear. We investigated the role of ESWT on the early proinflammatory response using a severe, full-thickness and highly inflammatory cutaneous burn wound in a murine model. Various wound-healing parameters were measured and leukocyte infiltration quantitated. A panel of 188 candidate genes known to be involved in acute inflammation and wound healing was screened. We show that ESWT of burn wounds 1 hour postwounding significantly blunts polymorphonuclear neutrophil and macrophage infiltration into the wound. ESWT treatment potently attenuates both CC- and CXC-chemokine expression, acute proinflammatory cytokine expression and extracellular matrix proteolytic activity at the wound margin. Given these findings and the clinical success of ESWT, we speculate that ESWT may be a potential therapeutic modality to treat severe wounds wherein excessive inflammatory responses involving increased levels of inflammatory cells, proinflammatory cytokines and proteases may become self-resolving allowing wound healing to progresses by way of normal physiological repair processes. PMID- 19291112 TI - Evaluation of the quality and cost-effectiveness of Versajet hydrosurgery. AB - Judicious resource allocation to maximise benefits is essential to health care decision-making, particularly with increasingly expensive treatments and rising populations. Accurate wound debridement is important, and while many methods exist, surgical debridement is currently the gold standard with material costs at approximately pound 140. Versajet, a hydrosurgical device, is suggested by some to have facilitated a paradigm shift in wound management by allowing debridement of undesirable tissue while accurately preserving viable structures. Its proponents claim that it minimises surgical duration, length of hospital admission, improves wound healing and may be deployed at the bedside or in the community. Consoles and single-use hand-units cost pound 6000- pound 7000 and pound 220- pound 240, respectively. Some propose that hydrosurgery may provide an efficacious, efficient and cost-effective alterative to surgical debridement. However, the evidence available is largely based on expert opinion. Methodological flaws of such descriptive studies include the lack of control groups, selection bias and lack of blinding. Hence, these claims need to be interpreted cautiously. This opinion review examines the available evidence as to whether Versajet hydrosurgery provides quality and cost-effective care. PMID- 19291113 TI - Silver resistance in MRSA isolated from wound and nasal sources in humans and animals. AB - Methicillin-resistant Staphylococcus aureus (MRSA) colonises skin, nasal passages and dermal wounds. Methods used to manage wounds infected and colonised with MRSA often include the use of topical antiseptics such as ionic silver and iodine. The objectives of this study were to determine the prevalence of silver-resistance (sil) genes in MRSA and methicillin-resistant coagulase-negative staphylococci (MR-CNS) isolated from wounds and nasal cavities of humans and animals, and also to determine the susceptibility of sil-positive and sil-negative MRSA isolates to a silver-containing Hydrofiber (SCH) wound dressing, on planktonic silE-positive and silE-negative MRSA. Polymerase chain reaction was used to determine the presence of three silver-resistance (sil) genes, silE, silP and silS in 33 MRSA and 8 methicillin-resistant staphylococci (MR-CNS). SilP and silS genes were absent in all isolates tested; however, two MRSA strains were found to contain the silE gene, together with one isolate of MR-CNS. Phenotypic resistance of the silE-positive strains and their susceptibility to the SCH dressing was evaluated using the zone of inhibition test on Mueller Hinton agar, and confocal laser microscopy using a live/dead fluorescent stain. Results confirmed that the SCH dressing was effective in killing all MRSA strains with and without the silE gene. First, this study showed that the prevalence of sil genes was low in the isolates investigated; and secondly, that the presence of a silver-resistance gene (silE) in MRSA and MR-CNS did not afford protection to the organism in the presence of a SCH wound dressing. The use of topical antiseptics in chronic wound care should be considered before the use of antibiotics that can result in their overuse and the risk of further resistance. PMID- 19291114 TI - Maggot debridement therapy with Lucilia cuprina: a comparison with conventional debridement in diabetic foot ulcers. AB - This is prospective case-control study of more than 18 months performed to assess the effectiveness of maggot debridement therapy (MDT) with the sterile larvae of Lucilia cuprina (a tropical blowfly maggot) for the treatment of diabetic foot ulcers. Literature thus far has only reported results with the temperate maggot, Lucilia sericata. This study documents outcome in diabetic foot wounds treated with maggot debridement versus those treated by conventional debridement alone. In this series of 29 patients treated with MDT, 14 wounds were healed, 11 were unhealed and 4 were classified under others. The control group treated by conventional debridement had 30 patients of which 18 wounds were healed, 11 unhealed and 1 classified under others. There was no significant difference in outcome between the two groups. The conclusion that can be made from this study is that MDT with L. cuprina is as effective as conventional debridement in the treatment of diabetic foot ulcers. It would be a feasible alternative to those at high risk for surgery or for those who refuse surgery. PMID- 19291115 TI - Haemodynamic effects of -75 mmHg negative pressure therapy in a porcine sternotomy wound model. AB - Previous research has shown -125 mmHg to be the optimal negative pressure for creating an environment that promotes wound healing, and this has therefore been adopted as a standard pressure for patients with deep sternal wound infection. However, it has not yet been clearly shown that -125 mmHg is the optimal pressure from a haemodynamic point of view. Furthermore, there have been reports of cardiac rupture during -125 mmHg negative pressure therapy. We therefore studied the effects of a lower pressure: -75 mmHg. Twelve pigs were used. After median sternotomy, sealed negative pressure therapy of -75 mmHg was applied. Baseline measurements were made and continuous recording of the cardiac output, end-tidal CO(2) production, mean arterial pressure, mean pulmonary pressure (pulmonary artery pressure), systemic vascular resistance, pulmonary vascular resistance, left atrial pressure and central venous pressure was started. Six pigs served as controls. No statistically significant difference was observed in any of the haemodynamic parameters studied, compared with the controls. The present study shows that, with a suitable foam application technique, -75 mmHg can be applied without compromising the central haemodynamics. PMID- 19291116 TI - Development of vaginal erosion and vesicocutaneous fistula following midurethral transvaginal tape with PelviLace for treatment of stress urinary incontinence. AB - Stress urinary incontinence (SUI) is a common diagnosis that greatly impacts quality of life for many women. Current therapies implement the use of suburethral slings, either synthetic or xenografts, to remedy this condition. In this study, we report on a patient with SUI, treated with PelviLace Biourethral Support, who suffered multiple issues postoperatively. PMID- 19291117 TI - Conservative management of ulcerated haemangioma--twenty years experience. AB - Ulceration is the most common complication of haemangioma and occurs in 5-15% of cases. The present study was carried out to evaluate the clinical features of ulcerated haemangioma and efficacy of the management protocol adopted by us over a period of 20 years. All patients with ulcerated haemangioma were evaluated on the basis of age at presentation, sex, rural or urban distribution and site of haemangioma. Treatment included application of topical antibiotic and systemic antibiotic and analgesic for pain. The total number of patients was 608. The male to female ratio was 1: 2.28. The rural:urban distribution was 2.43:1. The most common site of involvement was head and neck. Mean age of patients was 5.60 +/- 2.44 months. Mean size of haemangioma and ulceration was 47.30 +/- 20.67 cm(2) and 7.49 +/- 4.52 cm(2), respectively. The mean time for ulcer healing was 40.06 +/- 19.41 days. Ulcer size of more than 10 cm(2) took more time to heal. Response to treatment was satisfactory. Ulcerated haemangioma usually occurs before completion of 1 year of age; hence, every patient with haemangioma needs careful attention. Adequate treatment and regular follow up brings satisfactory response in the patients. PMID- 19291118 TI - Fixed sporotrichosis as a cause of a chronic ulcer on the knee. AB - Chronic wounds are a major health care problem worldwide. Wound healing is a holistic endeavour that requires an accurate identification of the specific entities interfering with wound healing in a particular patient. We present a case of fixed sporotrichosis as the cause of a chronic ulcer in the knee. Although a culture of Sporothrix schenckii could not be obtained, a positive response to the sporotrichin skin test, a skin biopsy showed a suppurative granuloma and an adequate response to oral administration of potassium iodide confirmed the diagnosis. The identification and correction of the underlying aetiology of any chronic wound is the first and most important step to restore wound healing. PMID- 19291119 TI - Intralesional administration of epidermal growth factor-based formulation (Heberprot-P) in chronic diabetic foot ulcer: treatment up to complete wound closure. AB - Previous studies have shown that an epidermal growth factor-based formulation (Heberprot-P) can enhance granulation of high-grade diabetic foot ulcers (DFU). The aim of this study was to explore the clinical effects of this administration up to complete wound closure. A pilot study in 20 diabetic patients with full thickness lower extremity ulcers of more than 4 weeks of evolution was performed. Mean ulcer size was 16.3 +/- 21.3 cm(2). Intralesional injections of 75 microg of Heberprot-P three times per week were given up to complete wound healing. Full granulation response was achieved in all 20 patients in 23.6 +/- 3.8 days. Complete wound closure was obtained in 17 (85%) cases in 44.3 +/- 8.9 days. Amputation was not necessary in any case and only one relapse was notified. The most frequent adverse events were tremors, chills, pain and odour at site of administration and local infection. The therapeutic scheme of intralesional Heberprot-P administration up to complete closure can be safe and suitable to improve the therapeutic goal in terms of healing of chronic DFU. PMID- 19291120 TI - Bacteriotherapy with Lactobacillus plantarum in burns. AB - Bacterial colonisation and infection remain the major causes of delayed healing and graft rejection following burns. Topical treatment is necessary to reduce the incidence of burn wound infection. Silver sulphadiazine (SD-Ag) is an often used microbicidal agent. However, this treatment produces adverse reactions and side effects. On the basis of experimental data and clinical application of lactobacilli as probiotics, we performed this exploratory study to establish the effectiveness of bacteriotherapy with topical application of the innocuous bacteria Lactobacillus plantarum cultured in De Man, Rogosa and Sharpe medium to provide an alternative method for burn treatment using SD-Ag as a reference. These innocuous bacteria would compete with other bacteria that are wound pathogens and would modify the wound environment and promote tissue repair. Eighty burned patients from the Plastic Surgery and Burns Unit were grouped into infected (delayed) second- and third-degree and non infected (early) third-degree burns and treated with L. plantarum or SD-Ag. The proportion of patients with delayed second-degree burns was 0.71 for L. plantarum and 0.73 for SD-Ag (relative rate: -2.72%) with respect to the decrease in bacterial load (<10(5) bacteria/g of tissue), promotion of granulating tissue wound bed and healing. In early third-degree burns, the values were 0.75 for L. plantarum and 0.84 for SD Ag (relative rate: -1.07%) in preventing wound infection and promotion of granulation tissue, 0.90 in graft taking for both treatments (relative rate: 0%) and 0.75 for L. plantarum and 0.77 for SD-Ag (relative rate: -2.60%) in healing. In delayed third-degree burns, values were 0.83 for L. plantarum and 0.71 for SD Ag (relative rate: +16.90%) with respect to the decrease in the bacterial load (<10(5) bacteria/g of tissue) and providing a granulating tissue wound bed, 0.90 in graft taking for both treatments (relative rate: 0%) and 0.75 for L. plantarum and 0.64 for SD-Ag (relative rate: + 17.19%) in healing. Although the number of patients (between 12 and 15 per group) did not enable the application of a power statistical test, these results suggest that the L. plantarum treatment should be studied in greater depth and could be used as a valid alternative for the topical treatment of burns. PMID- 19291121 TI - Effect of a water-based no-sting, protective barrier formulation and a solvent containing similar formulation on skin protection from medical adhesive trauma. AB - Trauma to the skin from repeated removal of adhesive-based medical products can cause pain, anxiety, risk of secondary infections and additional health care costs. Skin barrier formulations are used to protect the integrity from such trauma. However, not all formulations are equally protective. We report the results of a randomised controlled study comparing a solvent-free (SF) formulation and a solvent-containing (SC) formulation to the skin of 12 healthy volunteers aged 18-55 years. Treatments were applied at baseline to two of the four test sites on the back of each subject and repeated for 5 days. Measurements of pain, discomfort, erythema and skin water loss were taken 24 hours after each application. The SF formulation is associated with lower mean scores for erythema (day 5, P < 0.05) and lower values for transepidermal water loss (day 5, P < 0.05) and redness (days 4 and 5, P < 0.05) when compared with either no treatment or daily treatment with a SC formulation. There were no significant differences between subject responses when pain on application of the test formulation or discomfort associated with removal of the medical adhesive tapes were rated. We conclude that a SF formulation provides better security against adhesive-derived skin trauma than a SC formulation. PMID- 19291123 TI - Incarcerated women and abortion provision: a survey of correctional health providers. AB - CONTEXT: Many women entering jail or prison are pregnant, and correctional facilities are therefore an important venue for providing a range of pregnancy related care, including access to abortion services. However, the availability of abortion services to inmates in the United States is unknown. METHODS: Between October 2006 and March 2007, surveys about abortion provision were mailed to 951 health professionals who provided clinical care in correctional facilities. Descriptive statistics were tabulated, and measures of association were calculated using chi-square analysis. RESULTS: Of the 286 respondents who returned analyzable surveys, 68% indicated that inmates at their facility can obtain "elective" abortions. Eighty-eight percent of this group indicated that their facility provides transportation, but only 54% said that they help arrange appointments. Responses did not differ by providers' individual or institutional characteristics. However, providers from states with a Republican-dominated legislature or with a Medicaid policy that severely restricted coverage for abortion were more likely to indicate that availability of abortion services was limited than were those whose state had a predominantly Democratic legislature or a Medicaid program that covered all or most medically necessary abortions. CONCLUSIONS: Although incarceration does not preclude women's need for access to abortion, full access to services is not available in all settings. Improving women's overall health care in correctional settings should include increasing the accessibility of abortion services. PMID- 19291124 TI - Taking be proud! Be responsible! To the suburbs: a replication study. AB - CONTEXT: An important phase of HIV prevention research is replicating successful interventions with different groups and in different settings. METHODS: Be Proud! Be Responsible!, a successful intervention originally targeting black urban males and carried out in nonschool settings, was presented in health classes at urban and suburban schools with diverse student bodies. A group-randomized intervention study, which included 1,357 ninth and 10th graders from 10 paired schools in a Midwestern metropolitan area, was conducted in 2000-2002. Half the schools received the intervention, and half received a general health promotion program. Students' reports of their sexual behavior and selected cognitive mediators were analyzed immediately following the programs and four and 12 months later. RESULTS: Compared with students who received the control curriculum, students exposed to the intervention reported significantly greater knowledge of HIV, other STDs and condoms; greater confidence in their ability to control sexual impulses, to use condoms and to negotiate the use of condoms; and stronger intentions to use condoms. Stratified analyses revealed that the strongest intervention impacts were on knowledge and efficacy among males and students attending suburban schools. The intervention had no impact on sexual initiation, frequency of intercourse or condom use. CONCLUSIONS: Schools are a logical and viable setting for the dissemination and acquisition of information about HIV, including prevention strategies. However, the behavioral impact of an intervention may not be easily transferable when the program is taught outside a carefully controlled, nonschool setting. PMID- 19291125 TI - They "miss more than anything their normal life back home": masculinity and extramarital sex among Mexican migrants in Atlanta. AB - CONTEXT: Gender has been recognized as a significant influence on sexual health behaviors. Labor migration presents an important context of vulnerability for sexual health. To understand how the context of migration affects risk-related practices, both cultural and social aspects of gender need to be explored. METHODS: In the quantitative part of a mixed-methods study conducted in 1999 in Atlanta, 187 Mexican migrant men were asked about their demographic characteristics; sexual history; migration motivations; substance use; social support; leisure-time activities; and ideas about masculinity, sexuality and marriage. Multivariate regression analyses were conducted to test the association between these domains and men's number of partners since their arrival in Atlanta. RESULTS: Number of partners was positively associated with owning a home in Mexico; number of trips back to Mexico; social network size; having had a sex worker as a partner; and going out dancing and to strip clubs on weekends (coefficients, 0.3-4.1). It was negatively associated with age, education, contact with social network members and feeling that sex is tied to emotional intimacy (-0.4 to -1.0). CONCLUSIONS: Programs must acknowledge and target migrant men's social networks and the spaces in which they may encounter risky sexual situations. Multilevel strategies, such as the development of more health enhancing community spaces and the promotion of safer sexual practices should form part of comprehensive efforts to reduce sexual risk among migrant men. PMID- 19291127 TI - Sexual insistence and disliked sexual activities in young adulthood: differences by gender and relationship characteristics. AB - CONTEXT: Because sexual negotiations within young adult couples have consequences for sexual and reproductive health, it is important to determine associations between relationship contexts and sexual insistence. METHODS: Bivariate and multiple logistic regression analyses were conducted on data from 4,469 young adults participating in Wave 3 of the National Longitudinal Study of Adolescent Health (2001-2002). Analyses examined predictors of respondents' having experienced sexual insistence and having repeatedly engaged in sexual behaviors they disliked in a current relationship of at least three months' duration. RESULTS: Seven percent of men and 8% of women had had unwanted sex at their partner's insistence. A significantly greater proportion of women than of men (12% vs. 3%) had engaged repeatedly in sexual activities they disliked, primarily fellatio and anal sex. Relationship characteristics were associated with sexual insistence, but gender was not. For example, female respondents who reported unreciprocated love for their partner had higher odds of reporting sexual insistence perpetration than those who reported that they and their partner loved each other (odds ratio, 3.9). Females were more likely than males to report repeated participation in disliked sexual activities (3.7); relationship characteristics were relatively unimportant for this outcome. CONCLUSIONS: Young adults of both genders may need education on the importance of accepting a partner's sexual desires and being sensitive to both a partner's unwillingness to engage in an activity and the true extent of a partner's dislike of certain activities. They may also need guidance on how to voice their own preferences and dislikes. PMID- 19291128 TI - Trends in family planning in Russia, 1994-2003. AB - CONTEXT: Although Russian women have adequate knowledge of modern contraceptives, their level of use of these methods has been low, and abortion rates remain relatively high. METHODS: In 1994-2003, sexually active women aged 18-49 were interviewed about their contraceptive use as part of the Russia Longitudinal Monitoring Survey. Trends in contraceptive use were examined. Multivariate analyses were conducted to identify characteristics associated with reliable contraceptive use (IUD use or consistent oral contraceptive use) in 1994 and 2003. RESULTS: In each year, about 25% of sexually active women had used no contraceptive method in the past month, and 20% had used traditional methods. Prevalence of barrier method use increased from 9% to 21% between 1994 and 2003, while that of IUD use declined from 34% to 21%. These changes were especially pronounced in Moscow and St. Petersburg, and among younger women. Common reasons for nonuse were irregular sexual relations (cited by 29% of nonusers in 2003), desire for pregnancy (22%), perceived inability to get pregnant (15%), feeling that contraceptives are uncomfortable or unpleasant (15%), health problems (11%) and the availability of abortion (6%). In 1994 and 2003, the odds of reliable contraceptive use were elevated among women with at least a secondary education (odds ratios, 1.5-1.7), and were reduced among smokers (0.6-0.7). CONCLUSIONS: Modern, effective contraceptive use has not increased among sexually active Russian women. Growing use of barrier contraceptives may reflect HIV awareness. Obstacles to effective contraceptive use, such as attitudes and health service factors, need further clarification. PMID- 19291129 TI - Contraceptive use and abortion among women of reproductive age in st. Petersburg, Russia. AB - CONTEXT: Although the characteristics associated with contraceptive use among Russian women have been studied, no large-scale research has been conducted on women's use of different contraceptive methods and abortion. METHODS: A random sample of 1,147 women aged 18-44 completed questionnaires at local women's clinics in St. Petersburg in 2003-2004. Chi-square tests were used to examine differences in selected characteristics among age-groups, and logistic regression was used to assess associations between these characteristics and the use of contraceptive methods at last intercourse and abortion history. RESULTS: Among women at risk of unintended pregnancy, six in 10 had used reliable contraceptives (the pill, the IUD or condoms) at last intercourse; 42% had used condoms. Women in the middle income level were more likely than women with lower income to have used the pill (odds ratio, 2.1); cohabiting women and those who had had children had lowered odds of using condoms (0.6 and 0.3-0.5, respectively). More than half of those surveyed reported having had an abortion. Characteristics associated with increased odds of having had an abortion included being 25 or older (2.2 3.5), cohabiting (2.9), having high income (1.7), having experienced first intercourse before turning 18 (2.2) and having used no contraceptive method at first sex (1.5). The factor that was most strongly associated with abortion was a woman's number of births (4.9-5.7). CONCLUSIONS: Educational programs that promote the consistent use of condoms, especially among young women, and family planning programs that reduce financial barriers to contraceptive use, are critically needed in Russia. PMID- 19291130 TI - Abortion access for incarcerated women: are correctional health practices in conflict with constitutional standards. PMID- 19291137 TI - Mapping of quantitative trait loci affecting behaviour in swine. AB - Behavioural indices in vertebrates are under genetic control at least to some extent. In spite of significant behavioural problems in farm animals, information on the genetic background of behaviour is sparse. The aim of this study was to map QTL for behavioural indices in swine under healthy conditions and after infection with Sarcocystis miescheriana, as behaviour can be significantly influenced by disease. This well-described parasite model subsequently leads to acute (day 14 p.i.), subclinical (day 28 p.i.) and chronic disease (day 42 p.i.), allowing the study and comparison of the behaviour of pigs under four different states of health or disease. The study was based on a well-described Pietrain/Meishan F(2) family that has recently allowed the detection of QTL for disease resistance. We have mapped six genome-wide significant and 24 chromosome wide significant QTL for six behavioural indices in swine. Six of these QTL (i.e. 20% of total QTL) showed effects on behavioural traits of the healthy pigs (day 0). Some of them (QTL on SSC11 and 18) lost influence on behavioural activities during disease, while the effects of others (QTL on SSC5, SSC8) partly remained during the whole experiment, although with different effects on the distinct behavioural indices. The disease model has been of high relevance to detect effects of gene loci on behavioural indices. Considering the importance of segregating alleles and environmental conditions that allow the identification of the phenotype, we conclude that there are indeed QTL with interesting effects on behavioural indices in swine. PMID- 19291138 TI - Radiation hybrid mapping of six porcine genes of the matrix metalloproteinase family. PMID- 19291139 TI - Genetic mapping of quantitative trait loci for resistance to Haemonchus contortus in sheep. AB - This paper presents results from a mapping experiment to detect quantitative trait loci (QTL) for resistance to Haemonchus contortus infestation in merino sheep. The primary trait analysed was faecal worm egg count in response to artificial challenge at 6 months of age. In the first stage of the experiment, whole genome linkage analysis was used for broad-scale mapping. The animal resource used was a designed flock comprising 571 individuals from four half-sib families. The average marker spacing was about 20 cM. For the primary trait, 11 QTL (as chromosomal/family combinations) were significant at the 5% chromosome wide level, with allelic substitution effects of between 0.19 and 0.38 phenotypic standard deviation units. In general, these QTL did not have a significant effect on faecal worm egg count recorded at 13 months of age. In the second stage of the experiment, three promising regions (located on chromosomes 1, 3 and 4) were fine mapped. This involved typing more closely spaced markers on individuals from the designed flock as well as an additional 495 individuals selected from a related population with a deeper pedigree. Analysis was performed using a linkage disequilibrium-linkage approach, under additive, dominant and multiple QTL models. Of these, the multiple QTL model resulted in the most refined QTL positions, with resolutions of <10 cM achieved for two regions. Because of the moderate size of effect of the QTL, and the apparent age and/or immune status specificity of the QTL, it is suggested that a panel of QTL will be required for significant genetic gains to be achieved within industry via marker-assisted selection. PMID- 19291140 TI - The management of patent ductus arteriosus in Australia and New Zealand. AB - AIM: This study aimed to establish current management practice for patent ductus arteriosus (PDA) among individual consultant neonatologists in Australia and New Zealand, to examine the influences that drives practice and highlight the importance of future randomised controlled trials in the region. METHODS: Eligible subjects were identified from the Directory of Neonatal Intensive Care Units in Australia and New Zealand, 2007. A questionnaire was sent online to each consultant and was followed up with a letter and telephone call. Seven questions addressed management approach, the drug used and the treatment regimen, threshold for referral for surgical ligation and the literature influencing practice. Data were collected from 22 August 2007 to 22 November 2007. RESULTS: The overall response rate was 95%. For infants < or =28 weeks or < or =1000 g, all consultants treat PDA by one of four distinct management approaches. Expectant management was favoured by 35%, echocardiographic targeted prophylaxis 32%, presymptomatic treatment 16% and prophylaxis by 17%. There were marked regional variations in practice. Within individual units, more than one approach is used in 14 out of 24 units. Long courses of indomethacin are used to treat PDA by 86%. For 22% of consultants, management is not influenced by published literature. CONCLUSIONS: Differences of opinion in the literature are reflected by the heterogeneity in clinical practice across regions and within units. Crucial questions undergoing evaluation are whether data extrapolated from a previous area are relevant to PDA in modern neonatology and whether targeting treatment early can translate to improved clinical outcome. PMID- 19291141 TI - New insights concerning methicillin-resistant Staphylococcus aureus disease. PMID- 19291142 TI - Laboratory tools and strategies for methicillin-resistant Staphylococcus aureus screening, surveillance and typing: state of the art and unmet needs. AB - The public health burden caused by methicillin-resistant Staphylococcus aureus (MRSA) infections is now widely recognized, and is a cause of public alarm. Effective MRSA risk management in the healthcare system as well as in the community should rely on accurate detection of reservoirs and sources of transmission, as well as on close monitoring of the impact of interventions on disease incidence and bacterial dissemination. MRSA carrier screening and disease surveillance, coupled with molecular typing, are key information tools for integrated MRSA control and individual risk assessment. These tools should be tailored to the distinct needs of local interventions and national prevention programmes. Surveillance schemes should primarily inform local staff and serve as quality assurance about MRSA risk management. New technologies, including the use of selective culture media and real-time PCR assays, allow faster detection of MRSA carriers upon admission or during stay in healthcare institutions. More research is needed to ascertain their cost-effectiveness for MRSA control. Likewise, tremendous progress has been made concerning molecular typing methods, with optimization and standardization of sequence-based technologies offering broad applicability and high throughput. However, no single S. aureus typing method is yet providing fully reliable information within the range of discrimination needed for public health action. Further refinement of genotyping methods and international harmonization of surveillance and typing schemes must be achieved to facilitate global MRSA control. PMID- 19291143 TI - Prevention and control of methicillin-resistant Staphylococcus aureus. AB - Recent efforts to combat infections have focused on pharmaceutical interventions. However, the global spread of antimicrobial resistance calls for the reappraisal of personal and institutional hygiene. Hygiene embodies behavioural and procedural rules that prevent bacterial transmission. Consequently, the chance of spreading bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) is significantly reduced. Hygiene is part of the primacy and totality of patient care, ensuring that no harm is done. Any prevention and control strategy must be underpinned by changes in attitude, embraced by all. The major components of preventing and controlling MRSA include hand and environmental hygiene (as part of standard precautions), patient isolation, and patient/staff decolonization. Improving hand hygiene practice is especially important where the risk of infection is highest, e.g. in intensive care. Physical isolation has two advantages: the physical barrier interrupts transmission, and this barrier emphasizes that precautions are required. With limited isolation facilities, risk assessment should be conducted to indicate which patients should be isolated. Environmental hygiene, although important, has a lower priority than standard precautions. When a patient is ready for discharge (home) or transfer (to another healthcare facility), the overall interests of the patient should take priority. All patients should be informed of their MRSA-positive status as soon as possible. Because of increased mupirocin resistance, a selective approach to decolonization should be taken. When MRSA-positive staff are identified, restricting their professional activity will depend on the nature of their work. Finally, politicians and others need to commit to providing the necessary resources to maximize MRSA prevention and control. PMID- 19291144 TI - Management of methicillin-resistant Staphylococcus aureus infections. AB - This review addresses selected aspects of the management of severe healthcare associated infections due to methicillin-resistant Staphylococcus aureus (MRSA), including the limitations of current therapy, potential alternative agents, new therapeutic options, clinical approaches to MRSA bacteraemia/endocarditis and ventilator-associated pneumonia, and strategies to improve outcomes in patients with severe MRSA infections. PMID- 19291145 TI - The structural and biochemical characterizations of a novel TET peptidase complex from Pyrococcus horikoshii reveal an integrated peptide degradation system in hyperthermophilic Archaea. AB - The structure of a 468 kDa peptidase complex from the hyperthermophile Pyrococcus horikoshii has been solved at 1.9 A resolution. The monomer contains the M42 peptidase typical catalytic domain, and a dimerization domain that allows the formation of dimers that assemble as a 12-subunit self-compartmentalized tetrahedron, similar to those described for the TET peptidases. The biochemical analysis shows that the enzyme is cobalt-activated and cleaves peptides by a non processive mechanism. Consequently, this protein represents the third TET peptidase complex described in P. horikoshii, thereby called PhTET3. It is a lysyl aminopeptidase with a strong preference for basic residues, which are poorly cleaved by PhTET1 and PhTET2. The structural analysis of PhTET3 and its comparison with PhTET1 and PhTET2 unravels common features explaining the general mode of action of the TET molecular machines as well as differences that can be associated with strong substrate discriminations. The question of the stability of the TET assemblies under extreme temperatures has been addressed. PhTET3 displays its maximal activity at 95 degrees C and small-angle neutron scattering experiments at 90 degrees C demonstrate the absence of quaternary structure alterations after extensive incubation times. In conclusion, PhTETs are complementary peptide destruction machines that may play an important role in the metabolism of P. horikoshii. PMID- 19291146 TI - Special gears for full-time engines: association of dystrophin-glycoprotein complex and focal adhesion complex with myosin heavy chain isoforms in rat skeletal muscle. PMID- 19291147 TI - Key functions of the energetic regulation are implicated in vitamin C action. PMID- 19291149 TI - There are three major families of crystallins: misnaming of alphaB crystallin. PMID- 19291150 TI - Effort-reward imbalance and burnout among German nurses in medical compared with psychiatric hospital settings. AB - The aim of this study was to investigate whether nurses' efforts and rewards, as well as the effort-reward imbalance (ERI) and burnout, differ between subjects working in psychiatric vs. medical hospitals and between nurses under education and examined nurses respectively. Furthermore, the relationship between ERI and burnout was evaluated. Nursing is associated with high levels of emotional strain and heavy workloads. Burnout and a negative ERI can result in high absenteeism and turnover and have been identified as reasons why nurses leave their profession. In the last decade, working conditions of the nursing profession have changed in Germany, but somatic and psychiatric hospitals developed in different ways. This development may lead to different profiles. A sample of 389 nurses (78.8% female) in four German hospitals was investigated. A total of 147 nurses worked in psychiatric hospitals and 236 nurses worked in medical (somatic) hospitals. Fifty participants were still under education. The Effort-Reward Imbalance Inventory measures effort, reward and overcommitment at job and provides an imbalance score between effort and reward. The Maslach Burnout Inventory with the subscales, emotional exhaustion, lack of accomplishment and depersonalization, was also used. Nurses working in medical hospitals reported more burnout and had higher ERI scores. Subjects under education were comparable to examined nurses in terms of burnout but had lower ERI scores. Multiple regression analyses showed all ERI scales to be significant predictors for emotional exhaustion, while age, field of work and educational status further predict effort or ERI respectively. At present, the working situation of nurses in different settings appears to be characterized by a perceived imbalance of effort and reward and is associated with a high risk of developing burnout symptoms. PMID- 19291151 TI - The measurement of psychotic acuity by nursing staff. AB - Accurate evaluation of patients' psychotic state is essential to decrease psychotic symptoms and protect the patient and others. The aim of this paper is to conduct a literature review in order to access the utility, reliability and validity of current rating scales that are purported to measure psychotic acuity of inpatient population. A search of a number of electronic databases was undertaken to retrieve potential articles that focus on the measurement of acute psychosis. We identified some conceptual and theoretical problems when using a scale that is monitoring the progress of discharge and assesses the outcome of treatments. The findings revealed a difficulty in finding a commonly agreed definition of acute psychosis and a problem of obtaining frequent measures, and the frequency of measurement and fluctuation in psychosis. The most dominant scales in assessing psychosis were reviewed: the Global Assessment of Functioning Scale, the Brief Psychiatric Rating Scale and the Positive and Negative Symptom Scale. Several issues related to the scales' inter-rater reliability and construct validity remain unexplored. None of these scales addressed the conceptual and theoretical problems that we identified. A new scale that will measure acuity of symptoms in inpatient settings needs to be created. PMID- 19291152 TI - Facing inadequacy and being good enough: psychiatric care providers' narratives about experiencing and coping with troubled conscience. AB - The aim of this study is to illuminate the meaning of encounters with a troubled conscience among psychiatric therapists. Psychiatric care involves ethical dilemmas which may affect conscience. Conscience relates to keeping or losing a sense of personal integrity when making judgments about one's actions. Ten psychiatric therapists were interviewed in June 2006. The interviews were tape recorded, transcribed verbatim and interpreted using a phenomenological hermeneutic method. Two themes 'Facing inadequacy' and 'Struggling to view oneself as being 'good enough'' are presented. In the therapists interviewed, awareness of their use of power, a sense of powerlessness and a sense of blame gave rise to feelings of betrayals and shameful inadequacy. By sharing their inadequacy with co-workers, they managed to endure the sense of their inadequacy which otherwise would have threatened to paralyse them. Finding consolation in sharing wearing feelings, becoming realistic and attesting their worthiness, they reached reconciliation and found confirmation of being good enough. The findings are interpreted in light of Logstrup's ethics of trust, according to which conscience alerts us to silent but radical ethical demand and the risk of self deception. PMID- 19291153 TI - ADHD in the college student: a block in the road. AB - Older adolescents with Attention Deficit Hyperactivity Disorder (ADHD) often struggle as they make the transition to college. Loss of parental supervision and structure, variable course schedules and the freedom and distractions of campus life can lead to health-risk behaviours for the college student with ADHD. The purpose of this study was to determine factors that help, as well as hinder, college students with ADHD as they learn to cope with everyday challenges of life and academics once they leave the structure and support of their parents' home. Investigators conducted semi-structured qualitative interviews with 15 college students with ADHD. Follow-up interviews confirmed and validated information obtained during initial interviews. In-depth qualitative analysis of transcribed interviews resulted in identification of three global themes: gaining insight about ADHD, managing life and utilizing sources of support. Each global theme contains factors that hinder, as well as factors that help the college student with ADHD. The factors that college students described as helping and hindering their transition to college can serve as the basis for interventions to educate and empower younger adolescents with ADHD well in advance of their transition to college. PMID- 19291154 TI - Survey of psychiatrists' views of nurse prescribing. AB - For mental health nurses and service users to realize the benefits of non-medical prescribing, psychiatrists need to understand the remit of this role and be actively involved in influencing it. Although healthcare policy in the UK supports the expansion of non-medical prescribing, it requires much more for the successful implementation of such initiatives in practice. The aim of this survey was to explore the attitudes of all grades of psychiatrist working in two mental health trusts in the West Midlands to nurse prescribing. A specifically designed questionnaire was sent out to two groups of psychiatrists to assess their attitudes towards non-medical prescribing. Psychiatrists (n = 147) completed and returned a specifically designed questionnaire of their views. The more senior doctors appeared less concerned about nurse prescribing. Junior doctors expressed equivocation towards the role, suggesting that nurse prescribers be consistently supervised and have limited access to mental health drugs. The findings from this study have considerable implications for teams if junior doctors hold different views to their senior colleagues. Without the assistance of trusts in facilitating role change, the introduction of new roles could potentially heighten conflict between professions. PMID- 19291155 TI - Monitoring oral health and dental attendance in an outpatient psychiatric population. AB - Impaired mental health has been associated with an increased need for dental care. Population surveys have indicated that people with enduring mental health problems make less frequent planned visits to the dentist and report a greater number of missing teeth than the general population. The study aims to examine oral health status, attendance to dental care, and medication in an outpatient psychiatric sample. A descriptive study combining a structured interview with a visual oral examination carried out in 113 outpatients under psychiatric care. Dental health was described in terms of sound, missing and/or filled teeth, and showed a relation between these categories and types of psychiatric diagnosis, age and numbers of antidepressant and neuroleptic drugs. Oral hygiene was found to be more neglected among men and in patients with the diagnosis of schizophrenia. The need of dental treatment was widespread, although regular dental visits were commonly reported. In order to maintain good oral health, regular dental check-ups should be encouraged for patients under psychiatric care. Further studies are required to reveal contributory causes for/to decreased oral health. The difficulty such individuals have in maintaining additional self efficacy raises questions about the necessity for oral health interventions in outpatient psychiatric services. PMID- 19291156 TI - Using and understanding consumer satisfaction to effect an improvement in mental health service delivery. AB - Consumer satisfaction is today, widely accepted as a measure of the level and quality of service received by consumers. The aim of this survey-based study is to explore consumer satisfaction with quality of care, staff, environment and discharge in a south eastern Sydney adult acute inpatient mental health unit. A cross-sectional analysis is pursued in order to identify aspects of the patient stay, which form an associative relationship with an overall rating of consumer satisfaction on a 10-point scale. During the survey period, there were 182 discharges. Seventy questionnaires (38.5%) were returned from this group. The survey results highlight a number of areas of identified need, enabling the service to prioritize organizational systems around meeting these needs. Multiple regression analysis identified three items in the survey, which were independently significant associates of overall consumer satisfaction. They included being happy with the service provided by the consumer support worker, having support for services on discharge and feeling safe and secure on the ward. The model containing these three items accounted for 50% of the variation in overall satisfaction. Two primary interventions have been developed because survey administration which, it is hoped, will address issues raised in the survey. The interventions were the development of an admission and discharge pathway and a ward-based psychosocial intervention programme, which includes the involvement of consumer support workers. PMID- 19291157 TI - A study of stigmatized attitudes towards people with mental health problems among health professionals. AB - The project aimed to assess stigmatized attitudes among health professionals directed towards patients with mental health problems. The Attitude to Mental Illness Questionnaire was used to assess participants' attitudes towards fictitious patients from a secure forensic hospital and patients with schizophrenia and substance use disorders. Participants were health professionals from acute and mental health settings. In total, 108 completed questionnaires were received. Participants had highly stigmatized attitudes towards patients from a forensic hospital and those with active substance use disorders. Attitudes were less stigmatized to people with substance use disorders who were recovering in remission. This suggested that health professionals have stigmatized attitudes towards an illness such as schizophrenia and this is worse towards patients from a secure hospital. The manner in which patients with substance use disorder are presented can have a significant effect on stigmatized attitudes by health professionals. PMID- 19291158 TI - User involvement in community mental health services--principles and practices. AB - Although user involvement has been the vision of mental health care for the last decades, there are indications of this vision remaining as rhetoric rather than reality in many clinical settings. The objective of this paper is to raise some fundamental questions associated with user involvement. Four challenges are examined: assumptions about the nature of the problems, the power of defining 'true knowledge', the power domination in service provision and community mental health care as an accommodating arena for maximizing the service user role. Finally, some strategies are suggested in order to overcome barriers towards realizing the intentions of user involvement. PMID- 19291159 TI - Locked doors in acute inpatient psychiatry: a literature review. AB - Many acute inpatient psychiatric wards in the UK are permanently locked, although this is contrary to the current Mental Health Act Code of Practice. To conduct a literature review of empirical articles concerning locked doors in acute psychiatric inpatient wards, an extensive literature search was performed in SAGE Journals Online, EBM Reviews, British Nursing Index, CINAHL, EMBASE Psychiatry, International Bibliography of the Social Sciences, Ovid MEDLINE, PsycINFO and Google, using the search terms 'open$', 'close$', '$lock$', 'door', 'ward', 'hospital', 'psychiatr', 'mental health', 'inpatient' and 'asylum'. A total of 11 empirical papers were included in the review. Both staff and patients reported advantages (e.g. preventing illegal substances from entering the ward and preventing patients from absconding and harming themselves or others) and disadvantages (e.g. making patients feel depressed, confined and creating extra work for staff) regarding locked doors. Locked wards were associated with increased patient aggression, poorer satisfaction with treatment and more severe symptoms. The limited literature available showed the urgent need for research to determine the real effects of locked doors in inpatient psychiatry. PMID- 19291160 TI - Thinking the unthinkable: does mental health nursing have a future? PMID- 19291161 TI - The Matrix Model of dual diagnosis service delivery. AB - The Matrix Model is essentially a strategy for managing dual diagnosis across a range of agencies. It is a way of implementing partnership working across services and commissioning structures. The Matrix Model was born out of hard experience at the coalface of dual diagnosis treatment at a tier four service in Bristol. A very common experience, which many may recognize, was that clients with complex mental health and addiction needs were being sent from 'pillar to post' in their treatment. Things needed to change. Here is a method of how things can change. Briefly, professionals in the drug/alcohol and mental health fields co-locate, working with clients in each other's workspaces. In doing this, they create nodes of integration. These nodes of integration link through parallel working to create a matrix. Outcome and key recommendation is that professionals in the drug/alcohol and mental health fields co-locate in each other's agencies, adopting an assertive outreach approach to working with dual diagnosis/complex needs clients. PMID- 19291162 TI - Emergency Room Outreach: a new approach. PMID- 19291165 TI - High thrombin-activatable fibrinolysis inhibitor levels may protect against recurrent fetal loss. PMID- 19291166 TI - False-positive tests for heparin-induced thrombocytopenia in patients with antiphospholipid syndrome and systemic lupus erythematosus. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is a severe complication of heparin therapy that can be associated with arterial or venous thrombosis and is caused by antibodies against platelet factor 4 (PF4)-heparin complex. Patients with antiphospholipid syndrome (APS) have been reported with positive tests for PF4-heparin complex antibodies by antigen assay. Whether such patients can be treated with heparin is a dilemma. OBJECTIVES: To determine the incidence and nature of the HIT immune reaction in patients with APS and/or systemic lupus erythematosus (SLE). METHODS: Antibodies against PF4-heparin complex were assayed by particle gel immunoassay (PaGIA), or enzyme immunoassay (EIA) with or without an excess of heparin. EIA for PF4 alone was also performed. Functional assays for HIT, that is, heparin-induced platelet activation (HIPA) and heparin-induced platelet aggregation, were also performed. RESULTS: In 32 of 42 patients (76.2%) with APS, APS and SLE, SLE, or SLE with antiphospholipid antibodies, EIA IgG or PaGIA for PF4-heparin complex antibodies were positive. Of these 32 samples, 26 (81.3%) tested positive for anti-PF4 antibodies. All 24 samples that were positive for PF4-heparin complex by EIA IgG were also positive for EIA IgG in the presence of heparin excess, and all were negative by the HIPA and heparin-induced platelet aggregation tests. CONCLUSION: A large proportion of patients with APS and/or SLE give false-positive HIT antigen test results that are presumably related to autoantibodies against PF4, which can be distinguished from true HIT antibodies by EIA for PF4-heparin complexes tested with heparin excess, and by functional assays. PMID- 19291167 TI - Abnormal platelet function in C3-deficient mice. AB - SUMMARY BACKGROUND AND OBJECTIVES: The complement system is a biochemical cascade composed of several plasma proteins that can interact with endothelial cells and blood cells, including platelets. In order to investigate the effect of the complement system on platelets, we studied platelet function in C3-deficient mice that lack complement activity. METHOD AND RESULTS: Tail-cut bleeding time was prolonged and platelet aggregation in response to protease-activated receptor-4 (PAR4) peptide was decreased in C3-deficient mice as compared with wild-type littermates. Platelet aggregation in response to other agonists (ADP and collagen) was similar between C3-deficient mice and their normal littermates. Isolated platelets from wild-type mice aggregate less in C3-deficient plasma than in normal plasma, and, conversely, addition of plasma from wild-type mice or plasma-purified C3 improved aggregation of C3-deficient platelets. We also monitored the formation of murine arteriole or venule thrombi in an intravital microscopy thrombosis model. We found that C3-deficient mice had a significantly delayed thrombotic response in arterioles as compared with their wild-type littermates. Furthermore, thrombi in C3-deficient mice were less stable and embolized more frequently than those in wild-type mice. CONCLUSIONS: Platelets of C3-deficient mice have subnormal function, resulting in a prolonged tail-cut bleeding time and delayed thrombosis after vessel wall injury. PMID- 19291168 TI - Effect of an expenditure cap on low-income seniors' drug use and spending in a state pharmacy assistance program. AB - OBJECTIVE: To estimate the impact of a soft cap (a ceiling on utilization beyond which insured enrollees pay a higher copayment) on low-income elders' use of prescription drugs. DATA SOURCES AND SETTING: Claims and enrollment files for the first year ( June 2002 through May 2003) of the Illinois SeniorCare program, a state pharmacy assistance program, and Medicare claims and enrollment files, 2001 through 2003. SeniorCare enrolled non-Medicaid-eligible elders with income less than 200 percent of Federal Poverty Level. Minimal copays increased by 20 percent of prescription cost when enrollee expenditures reached $1,750. RESEARCH DESIGN: Models were estimated for three dependent variables: enrollees' average monthly utilization (number of prescriptions), spending, and the proportion of drugs that were generic rather than brand. Observations included all program enrollees who exceeded the cap and covered two periods, before and after the cap was exceeded. PRINCIPLE FINDINGS: On average, enrollees exceeding the cap reduced the number of drugs they purchased by 14 percent, monthly expenditures decreased by 19 percent, and the proportion generic increased by 4 percent, all significant at p<.01. Impacts were greater for enrollees with greater initial spending, for enrollees without one of five chronic illness diagnoses in the previous calendar year, and for enrollees with lower income. CONCLUSIONS: Near-poor elders enrolled in plans with caps or coverage gaps, including Part D plans, may face sharp declines in utilization when they exceed these thresholds. PMID- 19291169 TI - Recognition of Candida albicans Als3 by the germ tube-specific monoclonal antibody 3D9.3. AB - Monoclonal antibody 3D9.3 (MAb 3D9.3) reacts with the surface of Candida albicans germ tubes and recognizes a protein epitope. We used a two-step chromatography procedure to purify and identify the antigen (3D9) from C. albicans strain 66396 germ tubes. MAb 3D9.3 recognized two intense protein bands at 140 and 180 kDa. A comparative analysis between theoretical and experimental mass spectrum peaks showed that both bands corresponded to Als3. This conclusion was supported by lack of reactivity between MAb 3D9.3 and an als3Delta/als3Delta mutant strain, and the fact that an immunoglobulin preparation enriched for Als3 specificity recognized the purified 3D9 antigen. PCR demonstrated that C. albicans strain 66396 has two different-sized ALS3 alleles that correspond to the two purified protein bands. Strain- and species-specificity of the 3D9 epitope were studied with various C. albicans strains and Candida species, such as closely related Candida dubliniensis. The 3D9 epitope was detected only in C. albicans, demonstrating the utility of MAb 3D9.3 for differentiation between C. albicans and C. dubliniensis. Adhesion assays demonstrated that MAb 3D9.3 blocks adhesion of C. albicans germ tubes to human buccal epithelial cells and vascular endothelial cells. PMID- 19291170 TI - Suppressive effects of Bifidobacterium longum on the production of Th2-attracting chemokines induced with T cell-antigen-presenting cell interactions. AB - In human trials, Bifidobacterium longum BB536 alleviates subjective symptoms of Japanese cedar pollinosis, an IgE-mediated type I allergy caused by exposure to Japanese cedar, and significantly suppresses the increase of plasma thymus- and activation-regulated chemokine (TARC) associated with pollen dispersion. In the present study, we investigated the suppressive effects of BB536 on the production of T helper type 2 (Th2)-attracting chemokines, such as TARC and macrophage derived chemokine (MDC), together with the mechanisms of their production. Murine splenocytes were cultured with heat-killed BB536, and the levels of Th2 attracting chemokines in the supernatants were measured. TARC and MDC were produced in cultures without stimulation, and the production was significantly suppressed by BB536. These chemokines were produced by antigen-presenting cells (APCs) of splenocytes stimulated with an anti-CD40 antibody. Furthermore, TARC production was induced with granulocyte macrophage colony-stimulating factor that was produced by T cells and dendritic cells. BB536 suppressed MDC production induced with the anti-CD40 antibody by APCs from the spleen, mesenteric lymph nodes (MLNs) and Peyer's patches, and it suppressed TARC production by APCs from the spleen and MLNs. These results indicate that BB536 suppresses the production of Th2-attracting chemokines induced by the T cell-APC interaction, suggesting a novel mechanism for alleviating symptoms of allergic disorders by probiotics. PMID- 19291171 TI - Mucosal vaccination with a multicomponent adenovirus-vectored vaccine protects against Streptococcus pneumoniae infection in the lung. AB - Streptococcus pneumoniae is a major bacterial respiratory pathogen. Current licensed pneumococcal polysaccharide and polysaccharide-protein conjugate vaccines are administered by an intramuscular injection. In order to develop a new-generation vaccine that can be administered in a needle-free mucosal manner, we have constructed early 1 and 3 gene regions (E1/E3) deleted, replication defective adenoviral vectors encoding pneumococcal surface antigen A (PsaA), the N-fragment of pneumococcal surface protein A (N-PspA), and the detoxified mutant pneumolysin (PdB) from S. pneumoniae strain D39. Intranasal vaccination with the three adenoviral vectors (Ad/PsaA, Ad/N-PspA, and Ad/PdB) in mice resulted in robust antigen-specific serum immunoglobulin G responses, as demonstrated by an enzyme-linked immunosorbent assay. In addition, nasal mucosal vaccination with the combination of the three adenoviral vectors conferred protection against S. pneumoniae strain D39 colonization in mouse lungs. Taken together, these data demonstrate the feasibility of developing a mucosal vaccine against S. pneumoniae using recombinant adenoviruses for antigen delivery. PMID- 19291172 TI - Cytological findings in a primary GIST of the gallbladder. PMID- 19291173 TI - Fine needle aspiration cytology of palatine tonsils: a study of 112 consecutive adult tonsillectomies. AB - OBJECTIVE: To study fine needle aspiration cytology (FNAC) findings of tonsillar lesions with histological controls and to assess its role in the diagnostic evaluation of tonsillectomy specimens. METHODS: This study consisted of 112 cases that required tonsillectomy, comprising 55 (49.1%) men and 57 (50.9%) women. The ages ranged between 20 and 62 years. The clinical diagnosis in 101 cases was chronic tonsillitis, whereas 11 were suspected of neoplasia. FNAC was performed before tonsillectomy under general or local anaesthesia or on fresh specimens using a 21-G needle. The smears were stained using Wright-Giemsa and Papanicolaou methods. Histological examination was carried out on surgical specimens of all cases and, when required, immunohistochemistry was performed on histological sections. The diagnostic outcomes between FNAC and surgical biopsy were compared. RESULTS: In this study, 106 cases were diagnosed as chronic tonsillitis/follicular hyperplasia, four cases as non-Hodgkin's lymphoma, one as Hodgkin's lymphoma and one as monophasic synovial sarcoma. All malignant cases were diagnosed by FNAC, but synovial sarcoma was incorrectly diagnosed as squamous cell carcinoma. Five cases clinically suspected of neoplasia were correctly diagnosed as chronic tonsillitis on cytology. CONCLUSION: Tonsillar aspiration is a safe procedure and is useful in the evaluation of tonsillectomy specimens. However, ancillary tests on cytological material are often needed when neoplasia is suspected and would help clinical management and allow histological examination of cases diagnosed cytologically as lymphoma. PMID- 19291174 TI - Significance of collagenous and mucinous spherulosis in breast cytology specimens. AB - OBJECTIVE: Spherulosis of the breast is a rare but distinct benign morphological entity. As there are few cytological reports of breast spherulosis, the significance of spherulosis among cytological specimens is unclear. The objective was to document cytological aspects of spherulosis. METHODS: A total of 3491 consecutive breast fine needle aspiration cytology (FNAC) samples and 69 nipple discharge cytology samples were reviewed. Papanicolaou-stained slides with or without Romanowsky staining were analysed. The corresponding 1926 histological specimens were also reviewed. RESULTS: We detected 17 cases of collagenous spherulosis (CS) and/or mucinous spherulosis (MS) among 3560 breast cytology specimens (0.48%). All samples were from women, who varied in age from 22 to 69 years. CS and/or MS were present in 15 of 3491 FNAC specimens (0.43%) and in two of 69 nipple discharge cytology specimens (2.9%). Corresponding histological specimens were available for 14 of the 17 specimens. Of the 14 specimens, 12 consisted of intraductal papilloma, one of fibroadenoma, and one of fibrocystic change. There was no spherulosis among the 1251 cytological specimens of malignant diseases. CONCLUSIONS: Cytological evidence of spherulosis is a good indicator of intraductal papilloma. PMID- 19291175 TI - Fine needle aspiration cytology in the diagnosis of cervical tuberculosis and paracoccidioidomycosis. PMID- 19291176 TI - The evaluation of human papillomavirus genotyping in cervical liquid-based cytology specimens; using the Roche Linear Array HPV genotyping assay. AB - OBJECTIVE: To ascertain the usefulness of the Roche Linear Array human papillomavirus (HPV) genotyping assay for assessing HPV genotypes in liquid-based cytology (LBC) samples and to evaluate this methodology within a cytopathology laboratory. These tests are of importance as persistent infection with high-risk HPV genotypes is considered a causal factor in the development of cervical cancer. METHODS: A total of 175 cervical LBC samples were tested using the Roche Linear Array HPV genotyping test. The suitability of the assay use in routine cytopathology laboratory was considered. HPV genotypes were matched to the cervical cytology results, which included negative, borderline nuclear abnormalities, mild, moderate and severe dyskaryosis. RESULTS: The assay could be applied to screening samples with the combined result available at the reporting stage. There were no test failures. All samples used after cytological analysis had sufficient DNA for testing. The results were reproducible and easily read and there was concordance of results between biomedical scientists. The results of the assay showed co-infection with multiple HPV genotypes was common in both high grade and low-grade cytology samples. The percentage of HPV+ samples in the normal cytology samples (although in this grouping the number of samples was low) was 37%. In the cytology samples reported as severe dyskaryosis the HPV genotypes most commonly found were HPV16 and HPV51. CONCLUSION: The assay was able to detect multiple HPV infection with a wide range of genotypes in LBC samples sent for routine cytological analysis. It would be suitable for use in a cytopathology laboratory. The results of the assay show that the genotype profile has some variation from other geographical regions, and more work is needed to determine population prevalence, to ascertain the impact of the HPV vaccine, to evaluate test for cure and HPV triage management. PMID- 19291177 TI - Chronic blepharitis caused by Demodex folliculorum mites. PMID- 19291178 TI - Identification of paraxanthine as the most potent caffeine-derived inhibitor of connective tissue growth factor expression in liver parenchymal cells. AB - BACKGROUND: Recently, we identified hepatocytes as the major cellular source of profibrogenic connective tissue growth factor (CTGF/CCN2) in the liver. Based on reports of a hepatoprotective effect of coffee consumption, we were the first to provide evidence that caffeine suppresses transforming growth factor (TGF)-beta dependent and -independent CTGF expression in hepatocytes in vitro and in vivo, thus suggesting this xanthine-alkaloid as a potential therapeutic agent. AIM: This study aims at comparing the inhibitory capacities of caffeine and its three demethylated derivates paraxanthine, theophylline and theobromine on CTGF expression in hepatocytes and hepatic stellate cells (HSC). RESULTS: Our data suggest paraxanthine as the most important pharmacological repressor of hepatocellular CTGF expression among the caffeine-derived metabolic methylxanthines with an inhibitory dosage (ID)50 of 1.15 mM, i.e. 3.84-fold lower than what is observed for caffeine. In addition, paraxanthine displayed the least cell toxicity as proven by the water-soluble tetrazolium-1 cell vitality assay. However, caffeine or any of the metabolites did not inhibit CTGF expression in HSC. At the toxicological threshold concentration of 1 mM for paraxanthine, we observed an inhibition of hepatocellular CTGF synthesis by 44%, which was strongly reverted in the presence of the specific competitive cyclic adenosine monophosphate inhibitor Rp-adenosine 3',5-cyclic monophosphorothioate triethylammonium salt. Furthermore, CTGF protein expression induced by various concentrations of TGF-beta (0.13-1 ng/ml) is still reduced by, on average, 27%/45% in the presence of paraxanthine (1.25 mM/2.5 mM). CONCLUSION: Our data provide an evidence-based suggestion of the caffeine-derived primary metabolite paraxanthine as a potentially powerful antifibrotic drug by its inhibitory effect on (hepatocellular) CTGF synthesis. PMID- 19291179 TI - The missing stone. PMID- 19291180 TI - Early prediction of sustained virological response at day 3 of treatment with albinterferon-alpha-2b in patients with genotype 2/3 chronic hepatitis C. AB - BACKGROUND: Albinterferon-alpha-2b (albIFN) is a long-acting fusion polypeptide composed of albumin and IFN-alpha-2b. In a phase 2 study of albIFN 1500 mug q2wk or q4wk in patients with genotype 2/3 chronic hepatitis C, albIFN demonstrated sustained virological response (SVR) rates of 62-77% (intent-to-treat population). AIMS: To assess the association of initial viral kinetics during albIFN therapy with baseline factors and SVR prediction. METHODS: In all, 43 patients were treated with albIFN 1500 mug (q2wk/q4wk) plus ribavirin (RBV) 800 mg/day for 24 weeks. Hepatitis C virus (HCV)-RNA levels were measured by real time polymerase chain reaction, insulin resistance by homeostasis model assessment of insulin resistance (HOMA-IR) and serum albIFN levels by enyzme linked immunosorbent assay. Prediction analysis was performed in a per protocol 28-patient subset who were > or =80% adherent to albIFN/RBV and had HCV-RNA levels measured at treatment day 3. RESULTS: Day-3 HCV-RNA level and first-phase viral decline as well as second-phase slope of viral decline were significantly associated with SVR. In adherent patients, 82.1% had a day-3 viral load <4.2 log(10) IU/ml or first-phase decline >1.25 log(10) IU/ml, which was predictive of SVR, both positively (95.7%; sensitivity: 100%) and negatively (100%; specificity: 83.3%). As low first-phase decline was associated with a high pretreatment HOMA-IR index (P=0.004) and a low day-3 serum albIFN level (P=0.01). CONCLUSIONS: First-phase viral decline with albIFN/RBV was predictive of SVR in this study and may be modulated in part by IR. PMID- 19291181 TI - Thyroid dysfunction in primary biliary cirrhosis, primary sclerosing cholangitis and non-alcoholic fatty liver disease. AB - BACKGROUND/AIMS: Primary biliary cirrhosis (PBC) is frequently associated with autoimmune diseases, including thyroid disease, although it is uncertain that this association is higher than in other liver diseases. METHODS: We compared the prevalence and incidence of thyroid dysfunction (TD) in a series of patients with PBC (n=67) with patients with primary sclerosing cholangitis (PSC) (n=79) and non alcoholic fatty liver disease (NAFLD) (n=97) seen in a tertiary referral centre who had previously participated in clinical trials. RESULTS: At initial evaluation, prevalence of TD in PBC was 13% compared with 11% in PSC (P=0.71) and 25% in NAFLD (P=0.08). Incidence of TD was 2.9 patients per 100 person-years in PBC compared with 2.1 patients per 100 person-years in PSC (P=0.57) and 1.8 patients per 100 person-years in non-alcoholic liver disease (P=0.45). Older age, female gender, biochemical abnormalities and concurrent autoimmune disorders were not predictive of the development of TD. CONCLUSIONS: TD was unexpectedly as common in patients with PBC as in patients with PSC and NAFLD, yet significantly more common than expected in the general population. Further investigation of thyroid disease in PSC and NAFLD is warranted. PMID- 19291183 TI - Psychometric properties of MacNew Heart Disease Health-related Quality of Life Questionnaire: Brazilian version. AB - AIM: . This paper is a report of a study to test the psychometric properties of the Brazilian Version of the MacNew Heart Disease Health-related Quality of Life Questionnaire. BACKGROUND: Evidence of acceptable psychometric properties of the original MacNew questionnaire has been reported. However, the psychometric performance of the Brazilian version has not been extensively studied. METHOD: Between November 2004 and June 2005, 159 patients with myocardial infarction completed the Brazilian version of the instrument. Reliability was determined with Cronbach's alpha coefficient; convergent validity with Pearson's correlation between the Brazilian MacNew questionnaire and the SF-36; and the 'known-group' approach was used to test discriminant validity. Ceiling and floor effects and practicality were analyzed. FINDINGS: It took an average of 8.9 minutes to complete the instrument. A ceiling effect was detected in the subscales and global scores, with no floor effect. Cronbach's alpha coefficient ranged from 0.87 to 0.92. Scores on similar domains of the SF-36 and the MacNew questionnaire were correlated (r = 0.66; 0.78 and 0.53 for the physical, emotional, and social domains). Correlation between the dissimilar SF-36 mental health and MacNew physical domains was lower at 0.45; correlation between the dissimilar SF-36 physical functioning and MacNew emotional domain was 0.64. Individuals without regional ventricular dysfunction or symptoms had statistically significantly higher scores than those with regional ventricular dysfunction or symptoms. CONCLUSION: The Brazilian version of the MacNew questionnaire has acceptable levels of validity, reliability and practicality. Further studies are advisable to confirm its divergent validity and to test its responsiveness. PMID- 19291182 TI - Endothelin inhibits cholangiocarcinoma growth by a decrease in the vascular endothelial growth factor expression. AB - BACKGROUND: Endothelins (ET-1, ET-2, ET-3) are peptides with vasoactive properties interacting with ET(A) and ET(B) receptors. ET-1 inhibits secretin stimulated ductal secretion (hallmark of cholangiocyte growth) of cholestatic rats by interaction with ET receptors. AIM: The aims of the studies were to evaluate (i) the effect of ET-1 on cholangiocarcinoma growth in Mz-ChA-1 cells and nude mice and (ii) whether ET-1 regulation of cholangiocarcinoma growth is associated with changes in the expression of vascular endothelial growth factor-A (VEGF-A), VEGF-C, VEGF receptor-2 (VEGFR-2) and VEGFR-3. METHODS: We determined the expression of ET(A) and ET(B) receptors on normal and malignant (Mz-ChA-1) cholangiocytes and human cholangiocarcinoma tissue and the effect of ET-1 on the proliferation and expression of VEGF-A, VEGF-C (regulators of tumour angiogenesis) and its receptors, VEGFR-2 and VEGFR-3, in Mz-ChA-1 cells. In vivo, Mz-ChA-1 cells were injected into the flanks of athymic mice and injections of ET 1 or saline into the tumours were performed daily. The effect of ET-1 on tumour size, cell proliferation, apoptosis, collagen quantity and the expression of VEGF A and VEGF-C and VEGFR-2 and VEGFR-3 were measured after 73 days. RESULTS: Higher expression of ET(A) and ET(B) was observed in malignant compared with normal cholangiocytes. ET-1 inhibited proliferation and VEGF-A, VEGF-C, VEGFR-2 and VEGFR-3 expression of Mz-ChA-1 cells. Chronic ET-1 treatment decreased tumour volume, tumour cell proliferation and VEGF-A and VEGF-C expression but increased apoptosis and collagen tissue deposition compared with controls. CONCLUSIONS: Modulation of VEGF-A and VEGF-C (by ET-1) may be important for managing cholangiocarcinoma growth. PMID- 19291184 TI - Resident perspectives of the determinants of quality of life in residential care in Ireland. AB - AIM: This paper is a report of a study conducted to identify the determinants of quality of life for older people living in residential care, including exploration of mediating factors at personal and institutional levels and to construct a model of these. BACKGROUND: The quality of life of older people living in residential care is an under-researched area. The focus in the gerontology literature has been on quality of care rather than quality of life for these people. There is emerging consensus, however, that quality of life for older people is a complex, multidimensional concept, incorporating both subjective and objective elements, whether people live at home or in residential care. METHOD: A grounded theory study was conducted in 2005-2006. A total of 101 interviews were undertaken with older people across 12 sites, representing different types of residential care in Northern Ireland. The constant comparative technique was used to analyse data. FINDINGS: Four themes having an impact on the quality of life of residents were identified: ethos of care; sense of self and identity; connectedness; and activities and therapies. Mediating and facilitating and/or constraining factors for quality of life were also identified. The core category was conceptualized as 'maximizing potential'. CONCLUSION: Achieving optimal quality of life for older people in residential care will remain elusive unless all the factors which affect on it are taken into account by practitioners and policy-makers. Improving the quality of life of older people in residential care will require action in respect of all of the domains identified in this paper. PMID- 19291185 TI - Social isolation in older adults: an evolutionary concept analysis. AB - AIM: This paper is a report of an analysis of the concept of social isolation as experienced by older adults. BACKGROUND: Social isolation is a serious problem with older adults that is expected to increase in the future. Although strong correlations exist between social isolation and negative health consequences, few interventions exist. One contributing factor to this is a lack of conceptual clarity. DATA SOURCES: The CINAHL, PsycINFO and Medline databases and the Web of Science were searched using the key terms social isolation, social networks, older adults, elderly and social integration for publications between 1983 and 2007. REVIEW METHODS: An evolutionary concept analysis was undertaken. References were read in their entirety and analysed based on the summary of surrogate terms, related concepts, attributes, antecedents and consequences. FINDINGS: Five attributes were identified: number of contacts, feeling of belonging, fulfilling relationships, engagement with others and quality of network members. Antecedents included: lack of relationships, psychological barriers, physical barriers, low financial and resource exchange, and prohibitive environment. Roy's Adaptation Model is proposed as a theoretical context into which social isolation fits. CONCLUSION: Numerous definitions are used to describe social isolation, but they lack uniformity, consistency and clarity; this impairs the further development and refinement of the concept. This concept analysis can offer additional understanding and a starting point for future research related to social isolation. PMID- 19291186 TI - Decision-making for acutely ill nursing home residents: nurses in the middle. AB - AIM: This paper is a report of a study to generate a model of nursing behaviours and social processes inherent in decision-making for acutely ill nursing home residents. BACKGROUND: Most research concerning clinical decision-making in nursing homes focuses on the perspectives of doctors. Much less is known about the perspectives and actions of nurses with regard to decision-making, despite the centrality of their roles in nursing homes. METHOD: Grounded theory was used. Data were collected in 2004 in four nursing homes in the United States of America using in-depth, semi-structured interviews, 74 hours of non-participant observation and informal conversational interviews with key nursing staff involved in decision-making. FINDINGS: Nurses strive to create a plan of care acceptable to family members and doctors, consistent with wishes of residents and most comfortable for residents. A unifying theme of satisfying all sides emerged as representative of the negotiation strategies used by nurses to address these competing points of view. Four phases in this negotiation occurred: weighing the significance; notifying the family; feeling it out; and playing the middleman. The outcome was either a decision for life-prolongation or for palliative care. CONCLUSION: Decisions for nursing home residents are complex and involve weighing and balancing the interests and preferences of many concerned participants, including residents, families and doctors. This process requires skill in clinical judgment, communication and collaboration. PMID- 19291187 TI - Clinical decision-making for 'as needed' medications in mental health care. AB - AIM: This paper is a report of a study exploring the medical and nursing decision making process associated with the prescription and administration of 'as needed' psychotropic medication. BACKGROUND: The administration of 'as needed' psychotropic medications is a relatively autonomous component of a nurse's role, allowing for the capacity to administer psychotropic medications rapidly in acute situations or at the request of a patient. From the research evidence available to date, it is very difficult to determine how doctors and nurses make decisions about the prescription and administration of 'as needed' psychotropic medications. METHOD: A qualitative exploratory-descriptive study was undertaken to explore nurses and doctors decision-making surrounding the administration of pro re nata or 'as needed' psychotropic medications. Nineteen medical and nursing staff from three mental healthcare sites (acute, secure and rehabilitation) in Australia participated in semi-structured interviews in 2006. Thematic content analysis of the transcripts was conducted independently by two members of the research team and then merged to form the final themes. RESULTS: Four themes were identified in the data: decision-making processes; factors which influenced the administration and prescription of 'as needed' medication; individual protocols and improving practice. CONCLUSION: In-service education should be developed for mental health nurses on psychotropic medications and 'as needed' medications, and on the appropriate use of 'as needed' medications as a behaviour management strategy. Further, an extensive review of 'as needed' medication prescription and administration compared to best practice guidelines is needed. PMID- 19291188 TI - Humanism and its critiques in nursing research literature. AB - AIM: This paper raises for debate the issue of how humanist ideas have been taken up by nurse scholars, particularly in research literature. BACKGROUND: Many nurses from the mid-1970s onwards have described and promoted humanism as an appropriate philosophical basis for nursing practice and research. This has been partly in an attempt to sharply differentiate the profession from medicine, and later, managerialism, which have been represented as reductionist and failing to adequately respond to the whole patient. METHODS: A summary of definitions of humanism and critiques of humanism in broad philosophical literature is followed by an examination and critique of literature appearing in PubMed published within nursing scholarship from 1976 to 2007 which discusses or promotes humanism in nursing practice or research. FINDINGS: Writers have attempted to enhance the importance of nursing by associating it with the humanistic project of accepting responsibility for realizing our human potential. They have promoted a version of research which is qualitative and centres on understanding individual lifeworlds of research participants because of a strong valuing of the experiences and perspectives of the individual. Much of the literature on this topic describes this humanism in dualistic contrast to medico-scientific reductionism and objectivity. CONCLUSION: Some of the presentations of humanistic nursing lack rigour and can be seen as doing little more than reproducing professional ideology. Scholars and others in the field of nursing could take the trouble to submit these ideologies to proper scrutiny. PMID- 19291189 TI - Operating theatre nurses' perceptions of competence: a focus group study. AB - AIM: This paper is a report of a study exploring nurses' perceptions of the components of competence in the operating theatre. BACKGROUND: Competency Standards for operating theatre practice are used in some countries to guide clinical and professional behaviours. The need for competence assessment has been enshrined, but the conceptualization and agreement about what signifies competence in Operating Theatre has been lacking. METHODS: Three focus groups were conducted with 27 operating theatre nurses in three major metropolitan hospitals in Queensland, Australia. Interviews were audio taped and field notes were taken. Data were collected during 2008. Thematic analysis was performed. FINDINGS: From the analysis of the textual data, three themes were identified: 'coalescence of theoretical, practical, situational and aesthetic knowledge within a technocratic environment'; 'the importance of highly developed communication skills among teams of divergent personalities and situations'; and 'managing and coordinating the flow of the list'. CONCLUSION: These findings have identified that competence in respect to components of knowledge, teamwork and communication, and the ability to coordinate and manage are important and should be incorporated in operating theatre Competency Standards. Additionally, findings may assist in the development of an instrument to measure operating nurses' perceived competence. PMID- 19291190 TI - Knowing in nursing: a concept analysis. AB - AIM: This paper is a report of an analysis of the evolution of the concept of knowing in nursing. BACKGROUND: Following the seminal contribution of Carper in 1978, knowing has been discussed with increasing frequency in the nursing literature with reference to the development of nursing knowledge. Various patterns of knowing, as well as research focused on reflection on experience, have been the foundation for activities and research designed to improve practice as well as patient care in clinical, community, education, cultural and administrative settings. METHODS: Rodgers' evolutionary method of concept analysis provided the framework for this analysis. The data source was a search of literature published from 1978 to 2007. Three aims guided the analysis: to understand how the concept of knowing is used in nursing, how it is used in other health-related disciplines and how the concept has evolved into the current interpretation. DISCUSSION: Analysis revealed that knowing in nursing refers to a uniquely personal type of knowledge constructed of objective knowledge interfaced with the individual's subjective perspective on personal experience. Knowing is a dynamic process and a result of personal reflection and transformation as the individual lives and interacts in the world. Its antecedents are experience, awareness and reflection. The consequences are understanding, finding meaning and transformation. CONCLUSION: This analysis illuminates an area in which nursing has carved out its own niche in healthcare research. The concept of knowing in nursing practice and research reflects a focus on the individual experience of health and illness. PMID- 19291191 TI - Fall risk factors in older people with dementia or cognitive impairment: a systematic review. AB - AIM: This paper is a report of a review conducted to identify and summarize specific risk factors for falls in older people with dementia or cognitive impairments as documented by prospective or case-control studies. BACKGROUND: People with dementia have a doubled to threefold risk for falls, but the reasons for this have not yet been fully explained. Several integrative literature reviews discuss possible specific fall risk factors. However, there is lack of a systematic evaluation of studies. DATA SOURCES: The CINAHL, PubMed, EMBASE and PsychInfo databases were searched for the period between 1980 and May 2007. REVIEW METHODS: A systematic review was conducted. Cohort or case-control studies published in English or German were included if they investigated risk factors for falls or fall-related injuries in a sample consisting of participants with dementia or cognitive impairment. Two reviewers independently assessed study quality. RESULTS: Six prospective studies were included in the review. These differed concerning samples, settings, follow-up periods and examined variables. Therefore, meta-analysis was not possible. Eight categories of risk factors emerged: disease-specific motor impairments, impaired vision, type and severity of dementia, behavioural disturbances, functional impairments, fall history, neuroleptics and low bone mineral density. CONCLUSION: There is lack of sound studies examining fall risk factors in cognitively impaired elders. Well-known risk factors such as motor impairment show particular characteristics in people with dementia. In addition, behavioural disturbances contribute to their high risk for falls. Further prospective studies are needed. PMID- 19291192 TI - Pain assessment tools for unconscious or sedated intensive care patients: a systematic review. AB - AIM: This paper is a report of a systematic review describing instruments developed for pain assessment in unconscious or sedated intensive care patients. BACKGROUND: Intensive care patients who are unconscious or sedated are unable to communicate and therefore recognition and assessment of the pain is difficult. To assess these patients' pain, it is important to use a valid and reliable pain assessment tool. DATA SOURCES: A systematic bibliographical review was conducted, based on seven databases, covering the period from January 1987 to February 2007. A total of 1,586 abstracts was identified and reviewed, 58 papers were selected for full-text review and nine papers were included in the review. METHODS: Two researchers independently reviewed the abstracts and three reviewers extracted the papers. The included papers were evaluated using a quality assessment instrument previously developed to evaluate pain assessment tools. RESULTS: Five different pain assessment tools were identified that had been used with unconscious or sedated intensive care patients. All five instruments included behavioural indicators and three included physiological indicators. Their psychometric properties varied and it was not possible to deduce their clinical utility. CONCLUSION: All instruments were reasonably new. In most of them psychometric testing was in an early stage or even absent. Before any of the reported instruments can be chosen in preference to others, it is essential to test their validity, reliability and feasibility further. PMID- 19291193 TI - Research use in clinical practice - extent and patterns among nurses one and three years postgraduation. AB - AIM: This paper is a report of a study of nurses' research use in clinical practice one and three years postgraduation in Sweden. BACKGROUND: Internationally, learning to critically appraise and use research is an educational objective within nursing training, with the aim of promoting research use in nursing practice. The extent to which these skills is acquired and used among relatively newly graduated nurses is largely unexplored, however. METHOD: . A descriptive study was conducted in 2006 using a national longitudinal survey of two nursing cohorts one (n = 1,365) and three (n = 933) years postgraduation. The self-reported extent of instrumental, conceptual and persuasive research use was measured. Data were analysed using both variable- and pattern-oriented approaches based on cluster analysis. RESULTS: Research use was reported to occur in about half or fewer of the working shifts. In both samples, seven clusters of nurses with different research use profiles were identified. Clusters representing overall low and very low users in all three types of research use were predominant both at one (45.6%) and three (51.6%) years postgraduation, whereas clusters of nurses reporting overall high research use were uncommon. The proportion of very low users was larger 3 years after graduation than 1 year after graduation. CONCLUSIONS: The low extent of reported research use, raises the question of whether scientific perspectives included in nursing education are translated into clinical application. The pattern-oriented approach illustrates the complexity of research use and identification of typical research use profiles in specific contexts may have potential to guide interventions aimed at supporting evidence-based practice. PMID- 19291194 TI - Ethical care for older persons in acute care settings. PMID- 19291195 TI - Nursing and spirituality. AB - Those matters that are judged to be spiritual are seen as especially valuable and important. For this reason it is claimed that nurses need to be able to offer spiritual care when appropriate and, to aid them in this, nurse theorists have discussed the nature of spirituality. In a recent debate John Paley has argued that nurses should adopt a naturalistic stance which would enable them to employ the insights of modern science. Barbara Pesut has criticized this thesis, especially as it is applied to palliative care. This paper re-examines this debate with particular attention to the meaning of 'spirituality' and the justification for accepting spiritual and religious theories. It is argued that when we take into consideration the great diversity among religious and spiritual ideas, the lack of rational means of deciding between them when they conflict, and the practicalities of nursing, we find that a spiritual viewpoint is less useful than a naturalistic one, when offering palliative care. PMID- 19291196 TI - How can Rorty help nursing science in the development of a philosophical 'foundation'? AB - What can nurse scientists learn from Rorty in the development of a philosophical foundation? Indeed, Rorty in his 1989 text entitled Contingency, Irony, and Solidarity tantalizes the reader with debates of reason 'against' philosophizing. Forget truth seeking; move on to what matters. Rorty would rather the 'high brow' thinking go to those that do the work in order to make the effort useful. Nursing as an applied science, has something real that is worth looking at, and that nurse researchers need to think about. And as a profession built upon relationships, we should be thinking of the exchanges we have with those around us, of the contrasts in vocabularies used and of the contingencies involved, letting this launch us into our imaginings and areas of enquiry. The business of nurse researchers is to study what nurses do--how we care; Rorty would have us care. But, not to dismiss the reflective thinker as Rorty advocates for the self doubting ironist to continue to seek the final vocabulary, the ideal of what 'this' means, accepting this as the best to be offered at the time. As a science struggling to find foundation, we need only to look at what we do and value--as antifoundational as Rorty portrays himself, Rorty 'ironically' may have revealed a foundation for nursing science that is consistent with its path. PMID- 19291197 TI - Continuity of nursing and the time of sickness. AB - This paper explores the relationship between temporal continuity in nursing and temporal features of sickness. It is based on phenomenological and hermeneutical philosophy, empirical studies of sickness time, and the nursing theories of Nightingale, of Benner and of Benner and Wrubel. In the first part, temporal continuity is defined as distinct from interpersonal continuity. Tensions between temporal continuity and discontinuity are discussed in the contexts of care management, of conceptualisations of disease and of time itself. Temporal limitations to the methodological concept of situation are discussed. The main part of this paper explores nurses' possibilities to relate to their patients' time, and how temporal features of sickness may warrant temporal continuity of nursing. Three temporal characteristics of sickness are discussed: the immediacy of patients' suffering, the basic continuity of life through sickness and health care, and the indeterminism and precariousness of sickness. The timing of nursing acts is discussed. The paper explores how sickness is both part of the continuity of life, and threatens this continuity. It concludes that this tension is implicitly recognised in the temporal continuity of nursing, which allows for discontinuous and continuous aspects of sickness time. Nurses accordingly perceive the sick person's time at several levels of temporality, and distinguish highly complex temporal processes in their patients' trajectory. Temporal continuity provides the time, flexibility, and closeness for nurses to perceive and act into time dimensions of individual sickness. The paper shows that temporal continuity of nursing is grounded in temporal characteristics of severe sickness. It suggests that temporal continuity is an important theoretical concept in nursing. PMID- 19291198 TI - Towards an ontological theory of wellness: a discussion of conceptual foundations and implications for nursing. AB - In this article a discussion of the phenomenon of wellness and its relevance to contemporary nursing practice is developed. Drawing on phenomenology, the research literature and the author's own wellness research, an exposition of the concept of wellness is presented. It is proposed that the experience of being well is lived as a continuity of time and that it involves both a taking-for granted of the body and containment of the horizon of concern. The state of actually being well is also clarified and contrasted with the more common understanding of wellness as an optimal or future state. This discussion has significance for nursing knowledge development, in terms of our understanding of the experience of wellness and illness. It also has implications for how nurses approach their practice, particularly in the area of health promotion. PMID- 19291199 TI - Noddings's caring ethics theory applied in a paediatric setting. AB - Since the 1990s, numerous studies on the relationship between parents and their children have been reported on in the literature and implemented as a philosophy of care in most paediatric units. The purpose of this article is to understand the process of nurses' care for children in a paediatric setting by using Noddings's caring ethics theory. Noddings's theory is in part described from a theoretical perspective outlining the basic idea of the theory followed by a critique of her work. Important conceptions in her theory are natural caring (reception, relation, engrossment, motivational displacement, reciprocity) and ethical caring (physical self, ethical self, and ethical ideal). As a nurse one holds a duty of care to patients and, in exercising this duty, the nurse must be able to develop a relationship with the patient including giving the patient total authenticity in a 'feeling with' the patient. Noddings's theory is analysed and described in three examples from the paediatrics. In the first example, the nurse cared for the patient in natural caring while in the second situation, the nurse strived for the ethical caring of the patient. In the third example, the nurse rejected the impulse to care and deliberately turned her back to ethics and abandoned her ethical caring. According to the Noddings's theory, caring for the patient enables the nurse to obtain ethical insights from the specific type of nursing care which forms an important contribution to an overall increase of an ethical consciousness in the nurse. PMID- 19291200 TI - Compassion and professional care: exploring the domain. AB - Compassion unites people during times of suffering and distress. Unfortunately, compassion cannot take away suffering. Why then, is compassion important for people who suffer? Nurses work in a domain where human suffering is evidently present. In order to give meaning to compassion in the domain of professional care, it is necessary to describe what compassion is. The purpose of this paper is to explore questions and contradictions in the debate on compassion related to nursing care. The paper reviews classical philosophers as well as contemporary scientists' main arguments on compassion. First, I will examine the relationship between compassion and suffering. Second, how does one recognize serious suffering? This issue raises questions about the role of imagination and the need for identification. Third, literature describes compassion as an emotion. Some philosophers consider emotions uncontrollable feelings; others see a clear rational dimension in emotions. In order to determine what compassion is, it is necessary to weigh these contradictional arguments. Fourth, I will discuss motives for compassion. Is compassion an act of altruism or egoism? In this debate Nietzsche and Schopenhauer are well-known opponents. Today, analysis of their arguments leads to some surprising conclusions. Fifth, there is the issue of fault and compassion. Can we only feel compassionate when people who suffer are not to blame for their own suffering? Such a condition faces professional caretakers with a dilemma which needs a thorough analysis if compassion is to be of use in the field of professional care. Finally, I will explore the moral meaning of compassion. Compassion, described as a concept with cognitive as well as affective dimensions, also has volitional and behavioural aspects. These aspects specifically are of importance to nursing care and further research of compassion in the nursing domain. PMID- 19291201 TI - Scientism and the medicalization of existential distress: a reply to John Paley. PMID- 19291203 TI - The combined 12th International Philosophy of Nursing Conference and 15th New England Nursing Knowledge Conference. PMID- 19291205 TI - Neural markers of religious conviction. AB - Many people derive peace of mind and purpose in life from their belief in God. For others, however, religion provides unsatisfying answers. Are there brain differences between believers and nonbelievers? Here we show that religious conviction is marked by reduced reactivity in the anterior cingulate cortex (ACC), a cortical system that is involved in the experience of anxiety and is important for self-regulation. In two studies, we recorded electroencephalographic neural reactivity in the ACC as participants completed a Stroop task. Results showed that stronger religious zeal and greater belief in God were associated with less firing of the ACC in response to error and with commission of fewer errors. These correlations remained strong even after we controlled for personality and cognitive ability. These results suggest that religious conviction provides a framework for understanding and acting within one's environment, thereby acting as a buffer against anxiety and minimizing the experience of error. PMID- 19291206 TI - Issues in determining factors influencing bacterial attachment: a review using the attachment of Escherichia coli to abiotic surfaces as an example. AB - An understanding of the mechanisms which facilitate the attachment of Escherichia coli and other bacterial species to abiotic surfaces is desired by numerous industries including the food and medical industries. Numerous studies have attempted to explain bacterial attachment as a function of bacterial properties such as cellular surface charge, hydrophobicity and outer membrane proteins amongst others. Conflicting evidence in the literature both for and against a positive relationship may arise from the nature of the test methods used to measure them. A handful of recent studies utilizing technologies such as atomic force microscopy have begun to look at bacterial attachment at a single cell and molecular level. These studies may provide the information required to fully understand the underlying factors which influence bacterial cell attachment to abiotic surfaces. A number of issues in determining the influential factors of bacterial attachment have been identified from the literature: a lack of standardization and sensitivity of methods, as well as the value of measuring bulk properties of a number of cells rather than the behaviour of single cells which may overlook key interactions at a molecular level. These issues will need to be addressed in future studies in this area. PMID- 19291207 TI - Molecular detection of nifH gene-containing Paenibacillus in the rhizosphere of sorghum (Sorghum bicolor) sown in Cerrado soil. AB - AIMS: To develop a polymerase chain reaction (PCR)-based approach for the detection of nifH gene-containing Paenibacillus in environmental samples. METHODS AND RESULTS: The primers, nifHPAENf and nifHPAENr, were designed and tested with DNA from: (i) strains of different nitrogen-fixing Paenibacillus species, (ii) strains of other nitrogen-fixing genera and (iii) rhizosphere of sorghum sown in Cerrado soil amended with either 12 or 120 kg ha(-1) of nitrogen fertilizer. All nitrogen-fixing Paenibacillus strains tested and the DNA samples from rhizosphere soil were amplified when these primers were used, generating a 280 bp fragment. When the PCR products obtained from both sorghum rhizospheres were cloned and sequenced, the majority of the clones analysed could be identified as Paenibacillus durus. Moreover, a greater diversity in the nifH sequences could be observed in the rhizosphere treated with a high amount of nitrogen fertilizer. CONCLUSIONS: Nitrogen fertilization slightly influenced the structure of the nifH gene-containing Paenibacillus community in sorghum rhizospheres cultivated in Cerrado soil. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR detection method developed is adequate to assess the presence of nifH gene-containing Paenibacillus in the environment and can be used in future to determine the ecological role of this group of micro-organisms for the nitrogen input to the plants. PMID- 19291208 TI - In vitro synergistic effect of gentamicin with the anti-inflammatory agent diclofenac against Listeria monocytogenes. AB - AIMS: A total of nine Listeria monocytogenes strains (seven serotypes) were studied to ascertain whether the non-steroidal anti-inflammatory drug diclofenac (Dc) used in combination with the conventional antilisterial antibiotic gentamicin (Gm) or ampicillin (Am) synergistically augments the efficacy of the antibiotic in vitro. METHODS AND RESULTS: The effect of combination was evaluated by the checkerboard method to obtain a fractional inhibitory concentration (FIC) index followed by kill curves. Dc was synergistic with Gm (FIC 0.37) and there was indifference with Am (FIC 1) against L. monocytogenes ATCC 51774. The magnitude of the differences between killing by a single agent and the combination observed at 24 h was significant (P < 0.05) for Dc plus Gm but not Dc plus Am. CONCLUSIONS: Thus, the ability of extended antibiotic therapy may be improved with the help of this synergistic drug pair in listeriosis. SIGNIFICANCE AND IMPACT OF THE STUDY: Such findings may indicate parallel administration of anti-inflammatory and anti listeriosis drugs. PMID- 19291209 TI - Overview of diagnostic and detection methods for legionellosis and Legionella spp. AB - Since 1977, the diagnostic tools for Legionnaires' disease have been based on culture and serological investigations. Both methods require considerable time to produce results and have 'low' to 'reasonable' sensitivity. Since the introduction of urinary antigen tests in the mid 1990s, underdiagnosis has diminished and mortality has declined, thanks to early diagnosis. To obtain the most accurate diagnosis, culture, serological investigation, and urinary antigen testing should all be performed. In the last decade, advances in polymerase chain reaction technology allowed the development of assays detecting Legionella nucleic acids in clinical and environmental samples. Thus far, only widely varying results with relatively small series have been reported. Furthermore, these assays which are still labour intensive and complicated are not yet practicable for the average medical and/or environmental microbiological laboratory. PMID- 19291210 TI - Pseudomonas fluorescens and closely-related fluorescent pseudomonads as biocontrol agents of soil-borne phytopathogens. AB - Many strains of Pseudomonas fluorescens show potential for biological control of phytopathogens especially root pathogens. In taxonomic terms, several of them are indeed P. fluorescens sensu stricto, while others belong in fact to neighbouring species of the 'P. fluorescens' complex or to ill-defined related species within the fluorescent Pseudomonas spp. These bacteria have become prominent models for rhizosphere ecological studies and analysis of bacterial secondary metabolism, and in recent years knowledge on their plant-beneficial traits has been considerably enhanced by widening the focus beyond the case of phytopathogen directed antagonism. Current genomic analyses of rhizosphere competence and biocontrol traits will likely lead to the development of novel tools for effective management of indigenous and inoculated P. fluorescens biocontrol agents and a better exploitation of their plant-beneficial properties for sustainable agriculture. PMID- 19291211 TI - Benzoate-mediated changes on expression profile of soluble proteins in Serratia sp. DS001. AB - AIM: To assess differences in protein expression profile associated with shift in carbon source from succinate to benzoate in Serratia sp. DS001 using a proteomics approach. METHODS AND RESULTS: A basic proteome map was generated for the soluble proteins extracted from Serratia sp. DS001 grown in succinate and benzoate. The differently and differentially expressed proteins were identified using ImageMaster 2D Platinum software (GE Healthcare). The identity of the proteins was determined by employing MS or MS/MS. Important enzymes such as Catechol 1,2 dioxygenase and transcriptional regulators that belong to the LysR superfamily were identified. CONCLUSIONS: Nearly 70 proteins were found to be differentially expressed when benzoate was used as carbon source. Based on the protein identity and degradation products generated from benzoate it is found that ortho pathway is operational in Serratia sp. DS001. SIGNIFICANCE AND IMPACT OF THE STUDY: Expression profile of the soluble proteins associated with shift in carbon source was mapped. The study also elucidates degradation pathway of benzoate in Serratia sp. DS001 by correlating the proteomics data with the catabolites of benzoate. PMID- 19291212 TI - Development of a new method for the detection of lactic acid bacteria capable of protecting ham against Enterobacteriaceae. AB - AIMS: Challenge trials seem to be the best assessment approach to evaluate the potency of food protective cultures. However, this method is time consuming and often difficult to implement. Here, we describe the development of the 'sequential culturing method', a new method for the screening of strains as protective cultures. METHODS AND RESULTS: The sequential culturing method is based on the simulation, in a meat simulation medium (named BHI5L200), of the inhibition of Enterobacteriaceae by Lactobacillus, observed previously in situ. Results obtained with this sequential culturing method were in good agreement with those of the challenge test on sliced cooked ham and confirmed the antagonistic potency of Lactobacillus. The results obtained from the screening of 187 lactic acid bacteria (LAB) indicated that Lactobacillus sakei, Lactococcus lactis diacetylactis and Carnobacterium spp. were strong inhibitors of Enterobacteriaceae whereas Pediococcus spp., Leuconostoc spp., Weisselia spp. and other species of Lactobacillus and Lactococcus, did not possess the same inhibitory capacity. CONCLUSIONS: Sequential culturing method appeared to be a useful tool to rapidly select LAB cultures which are good candidates for bioprotection of meat. SIGNIFICANCE AND IMPACT OF THE STUDY: Sequential culturing method and simulating media could efficiently mimic challenge test experiments in the selection of potential protective culture for all types of food, on the condition to have the appropriate simulating media, corresponding to the food for which protective cultures were searched. PMID- 19291213 TI - Mathematical models of mycelium growth and ergosterol synthesis in stationary mould culture. AB - AIMS: To develop mathematical models for mycelium growth and ergosterol formation in conditions of periodic stationary culture; to verify possibilities of applying a model describing the relationship between ergosterol content and mycelium quantity. METHODS AND RESULTS: The mould growth and ergosterol formation models covering all phases of mould growth are described using a modified logistic equation with the addition of an exponential function. The correlation between ergosterol and mycelium biomass depended on the growth phase of mould. Meaningful relation was obtained for initial two phases, when both parameters were growing equally. The quadratic function for estimation of the biomass based on ergosterol content was formulated. The error resulting from the application of this function in initial phases of moulds growth was small at 5-7%, in the following phases it was at 11-31%. CONCLUSIONS: Mycelium biomass can be precisely determined basing on the ergosterol content, when we know the moulds growth phase. In natural environments, without the information about growth phases, it will be possible, but with the higher error. SIGNIFICANCE AND IMPACT OF THE STUDY: Presented model based on the ergosterol content making possible to estimate the quantity of mycelium in moulds cultures and natural environments. PMID- 19291214 TI - Brewer's spent grain and corn steep liquor as substrates for cellulolytic enzymes production by Streptomyces malaysiensis. AB - AIMS: To evaluate cellulase production by Streptomyces malaysiensis in submerged fermentation using brewer's spent grain (BSG) and wheat bran (WB) as carbon source, and corn steep liquor (CSL) as nitrogen source, as compared to yeast extract (YE), and partial characterization of the crude enzyme. METHODS AND RESULTS: Maximum cellulase production by Streptomyces malaysiensis (720 U l(-1)) occurred within 4 days incubation when using a growth medium containing BSG 0.5% (w/v) and CSL1.2% (w/v). CMCases activity showed to be stable over an acidic pH range (2.0-7.0) and in temperatures of 40-60 degrees C. Zymogram indicated three bands of CMCase activity, with different molecular masses. CONCLUSION: S. malaysiensis was able to grow and produce good levels of CMCases using solely brewer's spent grain and corn steep liquor as low-cost substrates, making this strain and these low cost by-product worthy for further investigation, and potentially feasible for biotechnological applications in different areas. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first study reporting the use of the low-cost by-products brewer's spent grain and corn steep liquor, as sole substrates for microbial enzyme production. PMID- 19291215 TI - Inactivation of Vibrio parahaemolyticus in pure culture, whole live and half shell oysters (Crassostrea virginica) by X-ray. AB - AIMS: To study the inactivation effect of different doses of X-ray on Vibrio parahaemolyticus in pure culture, inoculated whole live and half shell oysters and to evaluate the efficacy of X-ray doses on reduction of inherent microflora on oysters. METHODS AND RESULTS: X-ray was produced using RS 2400 generator system (Rad Source Technologies Inc.). Pure culture of V. parahaemolyticus, inoculated half and whole shell oysters with V. parahaemolyticus were treated with 0.0, 0.1, 0.5, 0.75, 1.0, 1.5, 2.0, 3.0 and 5.0 kGy X-ray. Surviving bacteria in the pure culture and inoculated oysters, before and after treatment, were enumerated using overlay plating (in TSA then TCBS) and most probable number (MPN) methods. A greater than 6.0 log reduction of V. parahaemolyticus was observed with 0.75, 2.0 and 5.0 kGy X-ray for pure culture, half shell and whole shell oysters, respectively. Treatment with 0.75, 2.0 and 5.0 kGy X-ray reduced the MPN to <3 for pure culture, half and whole shell oysters, respectively. Treatment with 1.0 kGy X-ray significantly (P < 0.05) reduced the inherent micro organisms on whole shell oysters from 4.7 +/- 0.1 to less than the detectable limit (<1.0 log CFU g(-1)). CONCLUSIONS: X-ray (1-5 kGy) significantly (P < 0.05) reduced V. parahaemolyticus and inherent microflora on oysters to less than detectable limit (<1.0 log CFU g(-1)). SIGNIFICANCE AND IMPACT OF THE STUDY: Treatment with X-ray could control pathogenic bacteria and extend the shelf life of oysters. PMID- 19291216 TI - Comparison of antimicrobial resistance patterns between clinical and sewage isolates in a regional hospital in Taiwan. AB - AIMS: To compare bacterial populations and antimicrobial resistance patterns between clinical and sewage isolates from a regional hospital in northern Taiwan. The dissemination of antibiotic-resistant bacteria from hospital compartments to the hospital sewage treatment plant was examined. METHODS AND RESULTS: A total of 1020 clinical isolates and 435 sewage isolates were collected between July and September 2005. The percentages of Gram-negative bacteria from the clinical and sewage isolates were 87.2% and 91.0%, respectively (P = 0.033). Escherichia coli were the leading bacterial isolates in both groups. Antimicrobial susceptibility testing showed a significant difference (P < 0.001) in resistance to ampicillin (85.6% vs 94.1%), ampicillin/sulbactam (31.7% vs 55.4%), cefazolin (29.2% vs 71.5%) and cefuroxime (20.7% vs 61.9%) between clinical and sewage coliform isolates, respectively. CONCLUSIONS: The sewage isolates had higher antimicrobial resistance rates than the clinical isolates from the same hospital. SIGNIFICANCE AND IMPACT OF THE STUDY: The low efficacy of the hospital sewage treatment may contribute to the dissemination of multidrug resistant bacteria from this hospital compartments to the environment. Practices which limit the disposal of antimicrobial agents into the wastewater system may be the possible measure to prevent the selection of multidrug-resistant bacteria from sewage treatment plants. PMID- 19291217 TI - Anti-Helicobacter pylori activity of Lactobacillus delbrueckii subsp. bulgaricus strains: preliminary report. AB - AIMS: To evaluate the activities of six Lactobacillus delbrueckii subsp. bulgaricus (LB) strains against 30 Helicobacter pylori strains by agar-well diffusion method. METHODS AND RESULTS: LB cultures [4 x 10(8)-4 x 10(9) CFU ml( 1)) either were prepared in milk at their native pH, 3.8-5.0, or were adjusted to pH 6.4-7.7. At low and neutralized pH, LB strains inhibited the growth by 40 86.7% and 16.7-66.7% of H. pylori strains, respectively. LB activity was strain dependent. At low and neutralized pH, one and five H. pylori strains, respectively, were not inhibited by any LB strain. LB2 and LB3, taken together, were active against most metronidazole and clarithromycin resistant strains. CONCLUSIONS: All LB strains inhibited a number of H. pylori strains, including also antibiotic resistant strains. LB activity was strain-dependent and better at low pH. At low pH values, the most active LB strains were LB1, LB2 and LB3, inhibiting 86.7% of H. pylori strains, while at neutralized pH values, the most active LB strains were LB2 and LB3, inhibiting 53.3 and 66.7% of H. pylori strains, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: LB could be utilized in the treatment or prophylaxis of H. pylori infection and warrants clinical investigations. PMID- 19291218 TI - Identification of a novel region of the GABA(B2) C-terminus that regulates surface expression and neuronal targeting of the GABA(B) receptor. AB - GABA(B) is a G protein-coupled receptor composed of two subunits, GABA(B1) and GABA(B2). GABA(B1) contains an endoplasmic reticulum-retention sequence and is trafficked to the cell surface only in association with GABA(B2). To determine whether the C-terminus of GABA(B2) regulates GABA(B) trafficking, we constructed forms of GABA(B2) with various C-terminal truncations and examined their surface expression. Truncation of GABA(B2) after residue 841 significantly reduced surface expression of both the subunit and the heterodimerized receptor. Turnover of the Delta841 construct, however, did not differ from that of full-length GABA(B2). To determine whether the C-terminus of GABA(B2) might target GABA(B) to neurites, cultured hippocampal neurons were transfected with the truncated GABA(B2) constructs. Truncation of GABA(B2) at residue 841 resulted in primarily somatic localization; furthermore, axonal trafficking of this construct was significantly more restricted than dendritic trafficking. Finally, to biochemically assess trafficking of the truncated GABA(B2) constructs, we digested transfected HEK293 cell lysates with endoglycosidase H. When GABA(B2) was truncated at residue 841, it became sensitive to digestion by this enzyme, indicating incomplete trafficking. Taken together, these data show that the region of the GABA(B2) C-terminus between residues 841 and 862 is important for regulating forward trafficking and neuronal targeting of the GABA(B) receptor. PMID- 19291219 TI - Changes in adult olfactory bulb neurogenesis in mice expressing the A30P mutant form of alpha-synuclein. AB - In familial and sporadic forms of Parkinson's disease (PD), alpha-synuclein pathology is present in the brain stem nuclei and olfactory bulb (OB) long before Lewy bodies are detected in the substantia nigra. The OB is an active region of adult neurogenesis, where newly generated neurons physiologically integrate. While accumulation of wild-type alpha-synuclein is one of the pathogenic hallmarks of non-genetic forms of PD, the A30P alpha-synuclein mutation results in an earlier disease onset and a severe clinical phenotype. Here, we study the regulation of adult neurogenesis in the subventricular zone (SVZ)/OB system in a tetracycline-suppressive (tet-off) transgenic model of synucleinopathies, expressing human mutant A30P alpha-synuclein under the control of the calcium/calmodulin-dependent protein kinase II alpha (CaMK) promoter. In A30P transgenic mice alpha-synuclein was abundant at the site of integration in the glomerular cell layer of the OB. Without changes in proliferation in the SVZ, significantly fewer newly generated neurons were observed in the OB granule cell and glomerular layers of A30P transgenic mice than in controls, most probably due to increased cell death. By tetracycline-dependent abrogation of A30P alpha synuclein expression, OB neurogenesis and programmed cell death was restored to control levels. Our results indicate that, using A30P conditional (tet-off) mice, A30P alpha-synuclein has a negative impact on olfactory neurogenesis and suppression of A30P alpha-synuclein enhances survival of newly generated neurons. This finding suggests that interfering with alpha-synuclein pathology can rescue newly generated neurons, possibly leading to new targets for therapeutic interventions in synucleinopathies. PMID- 19291220 TI - Towards a therapy for Huntington's disease (Commentary on Giampa et al.). PMID- 19291221 TI - Phosphodiesterase type IV inhibition prevents sequestration of CREB binding protein, protects striatal parvalbumin interneurons and rescues motor deficits in the R6/2 mouse model of Huntington's disease. AB - The phosphodiesterase type IV inhibitor rolipram increases cAMP response element binding protein (CREB) phosphorylation and exerts neuroprotective effects in both the quinolinic acid rat model of Huntington's disease (DeMarch et al., 2007) and the R6/2 mouse including sparing of striatal neurons, prevention of neuronal intranuclear inclusion formation and attenuation of microglial reaction (DeMarch et al., 2008). In this study, we sought to determine if rolipram has a beneficial role in the altered distribution of CREB binding protein in striatal spiny neurons and in the motor impairments shown by R6/2 mutants. Moreover, we investigated whether rolipram treatment altered the degeneration of parvalbuminergic interneurons typical of Huntington's disease (Fusco et al., 1999). Transgenic mice and their wild-type controls from a stable colony maintained in our laboratory were treated with rolipram (1.5 mg/kg) or saline daily starting from 4 weeks of age. The cellular distribution of CREB binding protein in striatal spiny neurons was assessed by immunofluorescence, whereas parvalbuminergic neuron degeneration was evaluated by cell counts of immunohistochemically labeled tissue. Motor coordination and motor activity were also examined. We found that rolipram was effective in preventing CREB binding protein sequestration into striatal neuronal intranuclear inclusions, sparing parvalbuminergic interneurons of R6/2 mice, and rescuing their motor coordination and motor activity deficits. Our findings demonstrate the possibility of reversing pharmacologically the behavioral and neuropathological abnormalities of symptomatic R6/2 mice and underline the potential therapeutic value of phosphodiesterase type IV inhibitors in Huntington's disease. PMID- 19291222 TI - Synchronous GABA-receptor-dependent potentials in limbic areas of the in-vitro isolated adult guinea pig brain. AB - Epileptiform discharges are known to reflect the hypersynchronous glutamatergic activation of cortical neurons. However, experimental evidence has revealed that epileptiform synchronization is also contributed to by population events mediated by GABA(A) receptors. Here, we analysed the spatial distribution of GABA(A) receptor-dependent interictal events in the hippocampal/parahippocampal region of the adult guinea pig brain isolated in vitro. We found that arterial perfusion of this preparation with 4-aminopyridine caused the appearance of glutamatergic independent interictal potentials that were reversibly abolished by GABA(A) receptor antagonism. Laminar profiles and current source density analysis performed in different limbic areas demonstrated that these GABA(A)-receptor mediated events were independently generated in different areas of the hippocampal/parahippocampal formation (most often in the medial entorhinal cortex) and propagated between interconnected limbic structures of both hemispheres. Finally, intracellular recordings from principal neurons of the medial entorhinal cortex demonstrated that the GABAergic field potential correlated to inhibitory postsynaptic potentials (membrane potential reversal, 68.12 +/- 8.01 mV, n = 5) that were interrupted by ectopic spiking. Our findings demonstrate that, in an acute seizure model developed in the adult guinea pig brain, hypersynchronous GABA(A)-receptor-mediated interictal events are generated from independent sources and propagate within limbic cortices in the absence of excitatory synaptic transmission. As spared or enhanced inhibition was reported in models of epilepsy, our data may support a role of GABA-mediated signaling in ictogenesis and epileptogenesis. PMID- 19291223 TI - Respiration-gated formation of gamma and beta neural assemblies in the mammalian olfactory bulb. AB - A growing body of data suggests that information coding can be achieved not only by varying neuronal firing rate, but also by varying spike timing relative to network oscillations. In the olfactory bulb (OB) of a freely breathing anaesthetized mammal, odorant stimulation induces prominent oscillatory local field potential (LFP) activity in the beta (10-35 Hz) and gamma (40-80 Hz) ranges, which alternate during a respiratory cycle. At the same time, mitral/tufted (M/T) cells display respiration-modulated spiking patterns. Using simultaneous recordings of M/T unitary activities and LFP activity, we conducted an analysis of the temporal relationships between M/T cell spiking activity and both OB beta and gamma oscillations. We observed that M/T cells display a respiratory pattern that pre-tunes instantaneous frequencies to a gamma or beta regime. Consequently, M/T cell spikes become phase-locked to either gamma or beta LFP oscillations according to their frequency range and respiratory pattern. Our results suggest that slow respiratory dynamics pre-tune M/T cells to a preferential fast rhythm (beta or gamma) such that a spike-LFP coupling might occur when units and oscillation frequencies are in a compatible range. This double-coupling process might define two complementary beta- and gamma-neuronal assemblies along the course of a respiratory cycle. PMID- 19291224 TI - Gamma activity and reactivity in human thalamic local field potentials. AB - Depth recordings in patients with Parkinson's disease on dopaminergic therapy have revealed a tendency for oscillatory activity in the basal ganglia that is sharply tuned to frequencies of approximately 70 Hz and increases with voluntary movement. It is unclear whether this activity is essentially physiological and whether it might be involved in arousal processes. Here we demonstrate an oscillatory activity with similar spectral characteristics and motor reactivity in the human thalamus. Depth signals were recorded in 29 patients in whom the ventral intermediate or centromedian nucleus were surgically targeted for deep brain stimulation. Thirteen patients with four different pathologies showed sharply tuned activity centred at approximately 70 Hz in spectra of thalamic local field potential (LFP) recordings. This activity was modulated by movement and, critically, varied over the sleep-wake cycle, being suppressed during slow wave sleep and re-emergent during rapid eye movement sleep, which physiologically bears strong similarities with the waking state. It was enhanced by startle eliciting stimuli, also consistent with modulation by arousal state. The link between this pattern of thalamic activity and that of similar frequency in the basal ganglia was strengthened by the finding that fast thalamic oscillations were lost in untreated parkinsonian patients, paralleling the behaviour of this activity in the basal ganglia. Furthermore, there was sharply tuned coherence between thalamic and pallidal LFP activity at approximately 70 Hz in eight out of the 11 patients in whom globus pallidus and thalamus were simultaneously implanted. Subcortical oscillatory activity at approximately 70 Hz may be involved in movement and arousal. PMID- 19291225 TI - Anti-Nogo-A antibody treatment promotes recovery of manual dexterity after unilateral cervical lesion in adult primates--re-examination and extension of behavioral data. AB - In rodents and nonhuman primates subjected to spinal cord lesion, neutralizing the neurite growth inhibitor Nogo-A has been shown to promote regenerative axonal sprouting and functional recovery. The goal of the present report was to re examine the data on the recovery of the primate manual dexterity using refined behavioral analyses and further statistical assessments, representing secondary outcome measures from the same manual dexterity test. Thirteen adult monkeys were studied; seven received an anti-Nogo-A antibody whereas a control antibody was infused into the other monkeys. Monkeys were trained to perform the modified Brinkman board task requiring opposition of index finger and thumb to grasp food pellets placed in vertically and horizontally oriented slots. Two parameters were quantified before and following spinal cord injury: (i) the standard 'score' as defined by the number of pellets retrieved within 30 s from the two types of slots; (ii) the newly introduced 'contact time' as defined by the duration of digit contact with the food pellet before successful retrieval. After lesion the hand was severely impaired in all monkeys; this was followed by progressive functional recovery. Remarkably, anti-Nogo-A antibody-treated monkeys recovered faster and significantly better than control antibody-treated monkeys, considering both the score for vertical and horizontal slots (Mann-Whitney test: P = 0.05 and 0.035, respectively) and the contact time (P = 0.008 and 0.005, respectively). Detailed analysis of the lesions excluded the possibility that this conclusion may have been caused by differences in lesion properties between the two groups of monkeys. PMID- 19291226 TI - Age matters. AB - The age of an experimental animal can be a critical variable, yet age matters are often overlooked within neuroscience. Many studies make use of young animals, without considering possible differences between immature and mature subjects. This is especially problematic when attempting to model traits or diseases that do not emerge until adulthood. In this commentary we discuss the reasons for this apparent bias in age of experimental animals, and illustrate the problem with a systematic review of published articles on long-term potentiation. Additionally, we review the developmental stages of a rat and discuss the difficulty of using the weight of an animal as a predictor of its age. Finally, we provide original data from our laboratory and review published data to emphasize that development is an ongoing process that does not end with puberty. Developmental changes can be quantitative in nature, involving gradual changes, rapid switches, or inverted U-shaped curves. Changes can also be qualitative. Thus, phenomena that appear to be unitary may be governed by different mechanisms at different ages. We conclude that selection of the age of the animals may be critically important in the design and interpretation of neurobiological studies. PMID- 19291227 TI - Perceptual stability during dramatic changes in olfactory bulb activation maps and dramatic declines in activation amplitudes. AB - We compared the concentration dependence of the ability of rats to identify odorants with the calcium signals in the nerve terminals of the olfactory receptor neurons. Although identification performance decreased with concentrations both above and below the training stimuli it remained well above random at all concentrations tested (between 0.0006% and 35% of saturated vapor). In contrast, the calcium signals in the same awake animals were much smaller than their maximum values at odorant concentrations <1% of saturated vapor. In addition, maps of activated glomeruli changed dramatically as odorant concentration was reduced. Thus perceptual stability exists in the face of dramatic changes in both the amplitude and the maps of the input to the olfactory bulb. The data for the concentration dependence of the response of the most sensitive glomeruli for each of five odorants was fitted with a Michaelis-Menten (Hill) equation. The fitted curves were extrapolated to odorant concentrations several orders of magnitude lower the smallest observed signals and suggest that the calcium response at low odorant concentrations is > 1000 times smaller than the response at saturating odorant concentrations. We speculate that only a few spikes in olfactory sensory neurons may be sufficient for correct odorant identification. PMID- 19291228 TI - Surround suppression sharpens orientation tuning in the cat primary visual cortex. AB - In the primary visual cortex (V1), the response of a neuron to stimulation of its classical receptive field (CRF) is suppressed by concurrent stimulation of the extraclassical receptive field (ECRF), a phenomenon termed 'surround suppression'. It is also known that the orientation tuning of V1 neurons becomes sharper as the size of the stimulus increases beyond the CRF. However, there have been few quantitative investigations of the relationship between sharpening of orientation tuning and surround suppression. We examined this relationship in 73 V1 neurons recorded from anesthetized and paralysed cats using sinusoidal grating patches as stimuli. We found that sharpening of orientation tuning was significantly correlated with the strength of surround suppression for large stimuli that cover both CRF and ECRF. Furthermore, simulation analysis using a variety of tuning widths and most suppressive orientation of orientation-tuned surround suppression demonstrated that broadly orientation-tuned surround suppression sharpens orientation tuning for large gratings without shift in optimal orientation. Our findings suggest that one of the functional roles of surround suppression in V1 is enhancement of orientation discrimination for large and uniformly patterned objects. PMID- 19291229 TI - Preference judgements involve a network of structures within frontal, cingulate and insula cortices. AB - Environmental stimuli constantly compete for human attention and in many cases decisions are made based on the affective meaning they convey. Although the network of structures involved in processing affective value has been well described, the specific contribution of these structures to the process by which affective value guides decision making is less well understood and is the focus of the present study. Thus, subjects read descriptions of individually tailored holidays, varying in incentive value and then made preference judgements, cognitive judgements or no decision. Choices made from an affective perspective, compared with those made from a cognitive perspective, activated a region of the anterior insula/operculum and also the anterior cingulate cortex. Furthermore, activity in perigenual, anterior cingulate cortex was correlated with subjective ratings of incentive value. In contrast, medial orbitofrontal cortex (OFC) and a region of posterior ventrolateral prefrontal cortex (PFC), bordering on the insula, were found to be more active when affective stimuli guided response selection than when no selection was made. However, only the activity in the ventrolateral PFC was specific to response selection based on affective compared with cognitive judgements. It is proposed that the necessary introspection required to make subjective preference judgements is provided by the insula and cingulate cortices, while the medial OFC and posterior ventrolateral PFC/insula cortices contribute to stimulus evaluation and motivational aspects of response selection, respectively. PMID- 19291230 TI - Effects of dopamine-related gene-gene interactions on working memory component processes. AB - Dopamine modulates complex cognitive functions like working memory and cognitive control. It is widely accepted that an optimal level of prefrontal dopamine supports working memory performance. In the present study we used a molecular genetic approach to test whether the optimal activity of the dopamine system for different component processes of working memory is additionally related to the availability of dopamine D2 receptors. We sought evidence for this assumption by investigating the interaction effect (epistasis) of variations in two dopaminergic candidate genes: the catechol-O-methyltransferase (COMT) Val(158)Met polymorphism, which has been shown to influence prefrontal dopamine concentration, and the DRD2/ANKK1-Taq-Ia polymorphism, which has been related to the density of D2 receptors. Our results show that COMT effects on working memory performance are modulated by the DRD2/ANKK1-TAQ-Ia polymorphism and the specific working memory component process under investigation. Val-participants- supposedly characterized by increased prefrontal dopamine concentrations- outperformed Val+ participants in the manipulation of working memory contents, but only when D2 receptor density could be considered to be high. No such effect was present for passive maintenance of working memory contents or for maintenance in the face of distracting information. This beneficial effect of a balance between prefrontal dopamine availability and D2 receptor density reveals the importance of considering epistasis effects and different working memory subprocesses in genetic association studies. PMID- 19291231 TI - Deficient responses from the lateral geniculate nucleus in humans with amblyopia. AB - Amblyopia or lazy eye is the most common cause of uniocular blindness in adults. It is caused by a disruption to normal visual development as a consequence of unmatched inputs from the two eyes in early life, arising from a turned eye (strabismus), unequal refractive error (anisometropia) or form deprivation (e.g. cataract). Animal models based on extracellular recordings in anesthetized animals suggest that the earliest site of the anomaly in the primate visual pathway is the primary visual cortex (corresponding to the striate cortex, cytoarchitectonic area 17 and area V1), which is where inputs from the two eyes are first combined in an excitatory fashion, whereas more distal and monocular processing structures such as the retina and lateral geniculate nucleus (LGN) are normal. Using high-field functional magnetic resonance imaging in a group of human adults with amblyopia, we demonstrate that functional deficits are first observable at a thalamic level, that of the LGN. Our results suggest the need to re-evaluate the current models of amblyopia that are based on the assumption of a purely cortical dysfunction, as well as the role for the LGN in visual development. PMID- 19291232 TI - Disruption of a mitochondrial MutS DNA repair enzyme homologue confers drug resistance in the parasite Toxoplasma gondii. AB - MutS homologues (MSHs) are critical components of the eukaryotic mismatch repair machinery. In addition to repairing mismatched DNA, mismatch repair enzymes are known in higher eukaryotes to directly signal cell cycle arrest and apoptosis in response to DNA-damaging agents. Accordingly, mammalian cells lacking certain MSHs are resistant to chemotherapeutic drugs. Interestingly, we have discovered that the disruption of TgMSH-1, an MSH in the pathogenic parasite, Toxoplasma gondii, confers drug resistance. Through a genetic selection for T. gondii mutants resistant to the antiparasitic drug monensin, we have isolated a strain that is resistant not only to monensin but also to salinomycin and the alkylating agent, methylnitrosourea. We have shown that this phenotype is due to the disruption of TgMSH-1 as the multidrug-resistance phenotype is complemented by a wild-type copy of TgMSH-1 and is recapitulated by a directed disruption of this gene in a wild-type strain. We have also shown that, unlike previously described MSHs involved in signalling, TgMSH-1 localizes to the parasite mitochondrion. These results provide the first example of a mitochondrial MSH that is involved in drug sensitivity and implicate the induction of mitochondrial stress as a mode of action of the widely used drug, monensin. PMID- 19291233 TI - Detection and quantification of Paenibacillus polymyxa in the rhizosphere of wild barley (Hordeum spontaneum) with real-time PCR. AB - AIM: To detect and quantify the plant drought tolerance enhancing bacterium Paenibacillus polymyxa in a collection of 160 Hordeum spontaneum rhizosphere samples at the 'Evolution Canyon' ('EC'), Israel. METHODS AND RESULTS: PCR primers and a FAM-TAMRA probe (6-carboxyfluorescein, 6-carboxy-tetramethyl rhodamine) targeting 16S rRNA genes were designed and used to detect and quantify the target strain. Two commercial kits, Bio101 Fast Spin and Mo Bio Ultra Clean Soil DNA, were tested for DNA isolation from the rhizosphere and surrounding soil. Population densities of P. polymyxa were studied in the rhizosphere of wild barley and surrounding soil from the contrasting climatic slopes at the 'EC' using the real-time PCR and culture based methods. CONCLUSION: Paenibacillus polymyxa is one of the best established species in wild barley rhizosphere at the 'EC' slopes. With the real-time PCR assay we are able to detect 1 pg of DNA per PCR corresponding to 100 cells per ml. The results at the 'EC' correlate well to bacterial estimations by culture based methods. SIGNIFICANCE AND IMPACT OF THE STUDY: Significantly higher P. polymyxa cell number was detected in the rhizosphere of arid 'African' microclimate indicating possible role of adaptive co-evolution with plants. PMID- 19291234 TI - Anhydrobiosis quotient: a novel approach to evaluate stability in desiccated bacterial cells. AB - AIM: The major objective of this study was the development of a methodology to quantify the anhydrobiotic ability of bacteria and its application to evaluate the stability of desiccated bacterial cells using the biocontrol agent Tsukamurella paurometabola C-924 as a model of anhydrobiote. METHODS AND RESULTS: Tsukamurella paurometabola C-924 was desiccated by spray-drying. Samples of desiccated cells were stored at several temperatures and viability and residual moisture were measured at different intervals of time. The term anhydrobiosis quotient (epsilon) was defined, and a scale of anhydrobiotic ability for classifying micro-organisms in terms of tolerance to desiccation was established (1 < or = epsilon < or = 15). The anhydrobiosis quotient was used to evaluate the stability of the anhydrobiotic cells. As a main result, changes in the anhydrobiosis quotient at several temperatures were fitted using a reparameterized Weibull model, which was found to be robust for the prediction of the stability at 4 degrees C. CONCLUSIONS: A novel methodology was developed to evaluate the desiccated state in bacteria. The anhydrobiosis quotient allows the quantitative estimation of the anhydrobiotic ability, and the mathematical model developed allows the prediction of the desiccated state of bacterial populations. SIGNIFICANCE AND IMPACT OF THE STUDY: The new methodology could be applied in studying the anhydrobiosis state of bacterial populations as a predictive tool for industrial and environmental microbiology. PMID- 19291235 TI - Development of real-time PCR tests for detecting botulinum neurotoxins A, B, E, F producing Clostridium botulinum, Clostridium baratii and Clostridium butyricum. AB - AIMS: To develop real-time PCR assays for tracking and tracing clostridia responsible for human botulism. METHODS AND RESULTS: Real-time PCR assays based on the detection of the genes ntnh encoding the nontoxin-nonhaemagglutinin (NTNH) proteins or the most homologous regions of the botulinum neurotoxin (bont) genes have been developed together with four real-time PCR assays, each being specific of the genes bont/A, bont/B, bont/E, bont/F and enables a toxin type-specific identification. The specificity of the assays was demonstrated using a panel of botulinum toxin producing clostridia (29 strains), nonbotulinum toxin producing clostridia (21 strains) and various other bacterial strains. The toxin type specific assays had a sensitivity of 100 fg-1000 fg of total DNA in the PCR tube (25-250 genome equivalents) which correspond to 10(3) to 10(4) cells ml(-1). After a 48 h enrichment in anaerobic conditions, these PCR assays allowed the detection of Clostridium botulinum type A in a naturally contaminated sample of 'foie gras' suspected in a C. botulinum outbreak. CONCLUSION: These PCR tests are specific and reliable for detection of heterogeneous BoNT producing clostridia responsible for human botulism. SIGNIFICANCE AND IMPACT OF THE STUDY: Adoption of these PCR assays is a step forward a reliable and rapid detection of these clostridia in food samples. PMID- 19291236 TI - Microbial population profiles of the microflora associated with pre- and postharvest tomatoes contaminated with Salmonella typhimurium or Salmonella montevideo. AB - AIMS: To determine the microflora profiles of pre- and postharvest tomatoes contaminated with Salmonella montevideo or S. typhimurium DT104. METHODS AND RESULTS: Salmonella montevideo or S. typhimurium was inoculated onto the flowers of tomato plants with the microflora of the subsequent fruit examined using a combination of Source Carbon Utilization and 16S rDNA-PCR profiling. From 16S rDNA profiles it was evident that tomatoes derived from Salmonella inoculated plants harboured a different microbial population compared to nontreated controls. The same result was observed for tomatoes inoculated at postharvest and subsequently stored for 14 days at 15 degrees C. From sequencing analysis it was found that tomatoes derived from Salmonella inoculated plants but testing negative for the enteric pathogen, frequently harboured Enterobacter and Bacillus spp. In contrast, both bacterial types were not found associated with tomatoes testing positive for Salmonella. CONCLUSIONS: Salmonella introduced onto tomatoes at pre- or postharvest alters the composition of the microbial community. The presence of Enterobacter and Bacillus spp negatively affects the persistence of Salmonella on preharvest tomatoes. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella appears to modify rather than become integrated into the microbial communities associated with tomatoes. Yet, the presence of antagonistic bacteria appears to reduce the persistence of the enteric pathogen. PMID- 19291237 TI - Arsenic-resistant bacteria isolated from agricultural soils of Bangladesh and characterization of arsenate-reducing strains. AB - AIMS: To analyse the arsenic-resistant bacterial communities of two agricultural soils of Bangladesh, to isolate arsenic-resistant bacteria, to study their potential role in arsenic transformation and to investigate the genetic determinants for arsenic resistance among the isolates. METHODS AND RESULTS: Enrichment cultures were performed in a minimal medium in the presence of As(III) and As(V) to isolate resistant bacteria. Twenty-one arsenic-resistant bacteria belonging to different genera of Gram-positive and Gram-negative bacteria were isolated. The isolates, with the exception of Oceanimonas doudoroffii Dhal Rw, reduced 2 mmol l(-1) As(V) completely to As(III) in aerobic conditions. Putative gene fragments for arsenite efflux pumps were amplified in isolates from Dhal soil and a putative arsenate reductase gene fragment was amplified from a Bacillus sp. from Rice soil. CONCLUSIONS: Phylogenetically diverse arsenic resistant bacteria present in agricultural soils of Bangladesh are capable of reducing arsenate to arsenite under aerobic conditions apparently for detoxification purpose. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides results on identification, levels of arsenic resistance and reduction of arsenate by the bacterial isolates which could play an important role in arsenic cycling in the two arsenic-contaminated soils in Bangladesh. PMID- 19291238 TI - Functionality of exopolysaccharides produced by lactic acid bacteria in an in vitro gastric system. AB - AIMS: To evaluate whether slime-exopolysaccharides (EPS) or capsular polysaccharide (CPS) production could protect the polymer-producing strains Streptococcus thermophilus CRL 1190 and Lactobacillus casei CRL 87 against the harsh conditions of an in vitro gastric system (GS). EPS stability on the GS was studied. METHODS AND RESULTS: An in vitro GS model containing human saliva and gastric juice was standardized. Polymer functionality on the cell viability and metabolic activity of the EPS-producing strains in the GS acidic conditions was evaluated. Two isogenic EPS/CPS deficient mutants were used for comparison. EPS or CPS conferred no significant protection on the cell viability of the studied strains after passage through the GS conditions. However, the phospho- and beta galactosidase activities of the EPS(+) strains were higher than those of the EPS( ). Cytoplasmic alterations in the wild-type and mutant strains and partial degradation of both EPS were detected. CONCLUSIONS: The presence of EPS/CPS protected the metabolic activity of the assayed LAB strains, but had no effect on survival at low pH. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of EPS/CPS as well as polymer resistance to the harsh conditions of the human GS could impact positively in probiotic strains to exert their properties in the host. PMID- 19291239 TI - Initial in vitro evaluations of the antibacterial activities of glucosinolate enzymatic hydrolysis products against plant pathogenic bacteria. AB - AIMS: The aim of the study was to evaluate the in vitro antibacterial effects of glucosinolate hydrolysis products (GHP) against plant pathogenic micro-organisms namely Agrobacterium tumefaciens, Erwinia chrysanthemi, Pseudomonas cichorii, Pseudomonas tomato, Xanthomonas campestris and Xanthomonas juglandis. METHODS AND RESULTS: Using a disc diffusion assay, seven different doses of 10 GHP were tested against each bacteria. The results showed that the isothiocyanates were potent antibacterials, whilst the other GHP were much less efficient. Moreover, the antibacterial effects were dose-dependent, increasing with the dose applied; 2-phenylethylisothiocyanate and sulforaphane showed the strongest inhibitory effects. The overall results show a great potential for using the isothiocyanates as an alternative tool to control undesired bacterial growth in plants. CONCLUSIONS: Glucosinolate hydrolysis products and more specifically the isothiocyanates: benzylisothiocyanate, 2-phenylethylisothiocyanate, the isothiocyanate Mix and sulforaphane, were effective phytochemicals against the in vitro growth of the phytopathogenic bacteria. The antibacterial activity exhibited by these phytochemicals reinforces their potential as alternatives to the traditional chemical control of phytopathogenic bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This current in vitro study is the first providing comparative data on GHP as potential control agents for plant pathogenic bacteria. However, more studies are needed to determine their possible allelopathic impacts e.g. inhibition of plant growth and negative effects on beneficial soil bacteria and fungi (mycorrhizae). PMID- 19291240 TI - The antimicrobial effects of glucosinolates and their respective enzymatic hydrolysis products on bacteria isolated from the human intestinal tract. AB - AIMS: The aim of the study was to evaluate the in vitro antibacterial activity of glucosinolates and their enzymatic hydrolysis product against bacteria isolated from the human intestinal tract. METHODS AND RESULTS: Using a disc diffusion bioassay, different doses of intact glucosinolates and their corresponding hydrolysis products were tested. There were clear structure-activity and concentration differences with respect to the in vitro growth inhibition effects as well as differences in the sensitivities of the individual bacteria. The most effective glucosinolate hydrolysis products were the isothiocyanates; sulforaphane and benzyl isothiocyanate were the best inhibitors of growth. Indole 3-carbinol had some inhibitory effects against the Gram-positive bacteria but had no effect, even at the highest dose, against the Gram-negative bacteria. Indole-3 acetonitrile had some inhibitory activity against the Gram-negative bacteria. Glucosinolates, nitriles and amines were ineffective at all the doses used. CONCLUSIONS: Glucosinolate hydrolysis products and specifically the isothiocyanates SFN and BITC have significant antimicrobial activity against Gram positive and Gram-negative bacteria, and might be useful in controlling human pathogens through the diet. SIGNIFICANCE AND IMPACT OF THE STUDY: This the first major in vitro study demonstrating the potential of these natural dietary chemicals as an alternative to, or in combination with, current antibiotic-based therapies for treating infectious diseases. PMID- 19291241 TI - Effects of modification of membrane lipid composition on Bacillus subtilis sporulation and spore properties. AB - AIMS: To determine effects of inner membrane lipid composition on Bacillus subtilis sporulation and spore properties. METHODS AND RESULTS: The absence of genes encoding lipid biosynthetic enzymes had no effect on B. subtilis sporulation, although the expected lipids were absent from spores' inner membrane. The rate of spore germination with nutrients was decreased c. 50% with mutants that lacked the major cardiolipin (CL) synthase and another enzyme for synthesis of a major phospholipid. Spores lacking the minor CL synthase or an enzyme essential for glycolipid synthesis exhibited 50-150% increases in rates of dodecylamine germination, while spores lacking enzymes for phosphatidylethanolamine (PE), phosphatidylserine (PS) and lysylphosphatidylglycerol (l-PG) synthesis exhibited a 30-50% decrease. Spore sensitivity to H(2)O(2) and tert-butylhydroperoxide was increased 30-60% in the absence of the major CL synthase, but these spores' sensitivity to NaOCl or Oxone was unaffected. Spores of lipid synthesis mutants were less resistant to wet heat, with spores lacking enzymes for PE, PS or l-PG synthesis exhibiting a two to threefold decrease and spores of other strains exhibiting a four to 10-fold decrease. The decrease in spore wet heat resistance correlated with an increase in core water content. CONCLUSIONS: Changing the lipid composition of the B. subtilis inner membrane did not affect sporulation, although modest effects on spore germination and wet heat and oxidizing agent sensitivity were observed, especially when multiple lipids were absent. The increases in rates of dodecylamine germination were likely due to increased ability of this compound to interact with the spore's inner membrane in the absence of some CL and glycolipids. The effects on spore wet heat sensitivity are likely indirect, because they were correlated with changes in core water content. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study provide insight into roles of inner membrane lipids in spore properties. PMID- 19291242 TI - Elaboration of an electroporation protocol for large plasmids and wild-type strains of Bacillus thuringiensis. AB - AIMS: To elaborate an effective electroporation protocol for large plasmids and wild type strains of Bacillus thuringiensis. METHODS AND RESULTS: The effect of DNA desalting, wall-weakening agency, cell growth conditions, electroporation solutions, and electric fields on electroporation efficiency was evaluated to optimize electroporation conditions for B. thuringiensis. By using this improved method, the greatest efficiency was reached 2 x 10(10 )CFU microg(-1) with pHT304, which is 10(4) times higher than previously reported. Four large plasmids (29.1, 44.9, 58 and 60 kb) were successfully transferred into the acrystalliferous B. thuringiensis strain BMB171; these results have not been achieved with previous protocols. Three wild type B. thuringiensis strains which could not be transformed previously were also transferred successfully. CONCLUSIONS: This improved method is more efficient for small plasmids; it is also appropriate for large plasmids and wild type B. thuringiensis strains which were not transformed by previous procedures. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study established an effective electroporation protocol for large plasmids and wild type strains of B. thuringiensis. This method is well suited for the cloning and expression of huge DNA fragments such as gene clusters in B. thuringiensis. It also can be used as a reference method for other Bacillus strains that are refractory to electroporate. PMID- 19291243 TI - Genomic diversity of vibrios associated with the Brazilian coral Mussismilia hispida and its sympatric zoanthids (Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi). AB - AIMS: A taxonomic survey of the vibrios associated with the Brazilian endemic coral Mussismilia hispida and the sympatric zoanthids (i.e. Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi). METHODS AND RESULTS: Mucus of 54 cnidarian specimens collected in three different places at Sao Sebastiao in two consecutive years (i.e. 2005 and 2006) was used for taxonomic characterization of the cnidarian microbiota. Ninety-eight of the 151 vibrio isolates fell within the vibrio core group according to partial 16S rDNA sequences. We performed the sequencing of recA and pyrH genes of all vibrio isolates. The most abundant taxa belonged to the vibrio core group (Vibrio harveyi, Vibrio rotiferianus, Vibrio campbellii and Vibrio alginolyticus), Vibrio mediterranei (=Vibrio shillonii) and Vibrio chagasii. With the exception of V. chagasii which was found only in the mucus of M. hispida, the other species appeared in different hosts with no evidence for the presence of host-specific clones or species. Using rep-PCR analysis, we observed a high genomic heterogeneity within the vibrios. Each vibrio isolate generated a different rep-PCR fingerprint pattern. There was a complete agreement between the grouping based on rep-PCR and concatenated sequences of pyrH, recA and 16S rDNA, but the pyrH gene has the highest discriminatory power for vibrio species identification. CONCLUSION: The vibrio core group is dominant in the mucus of these cnidarians. There is a tremendous diversity of vibrio lineages within the coral mucus. pyrH gene sequences permit a clear-cut identification of vibrios. SIGNIFICANCE AND IMPACT OF THE STUDY: The taxonomic resolution provided by pyrH (but not recA) appears to be enough for identifying species of vibrios and for disclosing putative new taxa. The vibrio core group appears to be dominant in the mucus of the Brazilian cnidarians. The overrepresentation of these vibrios may reflect as yet unknown ecological functions in the coral holobiont. PMID- 19291244 TI - Application of IHS criteria to headache attributed to spontaneous intracranial hypotension in a large population. AB - We applied the recent International Headache Society (IHS) criteria for headache related to spontaneous intracranial hypotension (SIH) to 90 consecutive patients with a final diagnosis of SIH confirmed by cerebral magnetic resonance imaging with contrast. Orthostatic headache (developing within 2 h of standing or sitting up) was present in 67 patients (75%) but appeared within 15 min after standing or sitting-as required by point A of the criteria-in only 53 (59%). Forty-four (49%) patients did not satisfy point A, including 22 (24%) with non-orthostatic headache and 14 (16%) with headache developing >or= 15 min after standing or sitting up; 80 (89%) did not satisfy point D. Only three (3%) patients had headache fully satisfying the IHS criteria. These findings indicate that the current IHS criteria do not capture most patients with SIH-associated headache. Excluding the requirement for response to epidural blood patch (criterion D) and considering headaches appearing within 2 h of sitting or standing up would capture more patients. PMID- 19291245 TI - Chronic migraine and medication overuse headache: clarifying the current International Headache Society classification criteria. AB - Despite the recent advances in the understanding and classification of the chronic daily headaches, considerable controversy still exists regarding the classification of individual headaches, including chronic migraine (CM) and medication overuse headache (MOH). The original criteria, published in 2004, were difficult to apply to most patients with these disorders and were subsequently revised, resulting in broader clinical applicability. Nonetheless, they remain a topic of debate, and the revisions to the criteria have further added to the confusion. Even some prominent headache specialists are unsure which criteria to use. We aimed to explain the nature of the controversies surrounding the entities of CM and MOH. A clinical case will be used to illustrate some of the problems faced by clinicians in diagnosing patients with chronic daily headache. PMID- 19291246 TI - Treat-early and treat-mild: role of fast vs. slow escalation of headaches. AB - This prospective, multi-center, observational study aimed to examine patients' early treatment decision process. Specifically, we assessed if the association between mild headache pain at treatment initiation and early treatment differed by the speed of headache escalation. Patients (n = 168) were instructed to collect information on their headache experience during the study period via an electronic diary over 30 consecutive days after enrollment. At the time of treatment, patients who treated early were 2.3 times as likely to experience mild headache pain as those who treated late. Controlling for the effect of escalation of headache, patients who treated early were three times as likely to report mild headache pain at dosing as those who treated late. The interaction between fast escalation of headache and mild pain was not statistically significant. Early treatment is associated with mild pain, regardless of the speed of headache escalation. PMID- 19291247 TI - Immediate response to greater auricular nerve blockade in red ear syndrome. PMID- 19291248 TI - Trigeminovascular wind-up and the pulsating nature of migraine pain. PMID- 19291250 TI - Mutagens manufactured in fungal culture may affect DNA/RNA of producing fungi. AB - Self-produced mutagens in culture by fungi may affect DNA analysis of the same fungi. This has not been considered previously. Many fungi produce numerous mutagenic secondary metabolites (SM) in culture. There is a paradox of growing fungi in media to produce representative DNA which also support mutagenic SM. This is a crucial issue in developing diagnostic and phylogenetic methods, especially for closely-related fungi. For example, idh gene analysis of the patulin metabolic pathway in fungi can be interpreted as producing some false negative and positive results in terms of possession, or nonpossession, of the gene from mutated strains. The most obvious mycotoxins and fungi to consider in this regard are aflatoxins and Aspergillus, as aflatoxins are the most mutagenic natural compounds. Many other fungi and SM are relevant. Conditions to grow fungi have not been selected to inhibit SM production although relevant data exist. In fact, fungi repair damaged nucleic acid (NA) and are capable of removing toxins by employing transporter proteins. These and NA repair mechanisms could be inhibited by secondary metabolites. Mutagenic effects may involve inhibition of DNA stabilizing enzymes. There may be an equivalent situation for bacteria. Researchers need to devise methods to reduce SM for valid protocols. More work on how mutagens affect the NA of producing fungus in vitro is required. The current review assesses the potential seriousness of the situation with selected papers. PMID- 19291251 TI - Rationale for the study methods and design of the epidemiology of lower urinary tract symptoms (EpiLUTS) study. AB - Accurate prevalence data on illnesses are of paramount importance for medical decision-making, healthcare planning, and the appropriate allocation of healthcare resources. When assessing the prevalence of any illness or health condition, it is important to base prevalence data on an assessment of the general population at large and not on selected groups of the population. Previous estimates of the prevalence of lower urinary tract symptoms (LUTS) have used postal surveys, telephone and in-person interviews, and, more recently, the Internet. Determining the mode of data collection for a research study is dictated by several factors, including target population, topic sensitivity, timeliness, and available resources. The purpose of this report is to document the rationale for conducting a large prevalence study of LUTS and its impact on patient outcomes using Internet- or Web-based surveys. A brief overview of important survey considerations is provided, and previous survey methods used in assessing the prevalence of LUTS discussed. The research objectives and sampling approach used in the EpiLUTS study in the USA, the UK and Sweden are presented, with a discussion of the advantages and disadvantages of this Internet-based sampling approach in relation to other epidemiological methods. PMID- 19291252 TI - Can fuzzy logic make things more clear? AB - Intensive care is a complex environment involving many signals, data and observations. Clinical decision support and artificial intelligence using fuzzy logic and closed loop techniques are methods that might help us to handle this complexity in a safe, effective and efficient way. Merouani and colleagues have performed a study using fuzzy logic and closed loop techniques to more effectively wean patients with sepsis from norepinephrine infusion. PMID- 19291253 TI - Darwin and Huxley revisited: the origin of allometry. AB - The relative sizes of parts of an organism frequently depend on the absolute size of the individual, a relationship that is generally described by power laws. I show here that these power laws are a consequence of the way evolution operates. PMID- 19291254 TI - Q&A: What did Charles Darwin prove? PMID- 19291255 TI - Evolutionary genomics and the reach of selection. AB - Unexpected findings in evolutionary genomics both question the role of selection in genome evolution and clarify how genomes work. PMID- 19291256 TI - Mayr, mathematics and the study of evolution. AB - In 1959 Ernst Mayr challenged the relevance of mathematical models to evolutionary studies and was answered by JBS Haldane in a witty and convincing essay. Fifty years on, I conclude that the importance of mathematics has in fact increased and will continue to do so. PMID- 19291258 TI - Ethics review: end of life legislation--the French model. AB - French law 2005-370 of April 22, 2005 (Leonetti's law) brings new rights to patients and clarifies medical practices regarding end of life care. This new law prohibits unreasonable obstinacy in investigations or therapeutics and authorizes the withholding or withdrawal of treatments when they appear "useless, disproportionate or having no other effect than solely the artificial preservation of life". Relief from pain is a fundamental right of patients. With regard to pain control, the law also allows doctors to dispense to patients "in an advanced or final phase of a serious and incurable affliction" anti-pain treatments as needed, even if these treatments, as a side effect, hasten their death. The drafting of advance directives regarding end of life constitutes a new right of patients. The decision to withdraw or withhold a treatment from a patient unable to express their will has to take into account the wishes they might have expressed through advance directives, and/or the wishes of a trusted person or, lastly, of the family. Before making any decision, physicians should respect a collegial medical procedure. Euthanasia defined as the act of terminating one's life on a patient's explicit request remains illegal. PMID- 19291259 TI - Estimating rates and patterns of morphological evolution from phylogenies: lessons in limb lability from Australian Lerista lizards. AB - Squamates (lizards and snakes) offer an exciting model system for research on the evolution of body form. A new phylogenetic study in BMC Evolutionary Biology of Australian lizards shows remarkable evolutionary lability in digit numbers among closely related species, but also highlights important challenges in this area. PMID- 19291260 TI - Why didn't Darwin discover Mendel's laws? AB - Darwin's focus on small quantitative variations as the raw material of evolution may have prevented him from discovering the laws of inheritance. PMID- 19291261 TI - Diabetes and sepsis outcomes--it is not all bad news. AB - Patients with diabetes mellitus have an increased risk of developing infections and sepsis. In this issue of Critical Care Esper and colleagues report on a large survey, involving 12.5 million sepsis cases, that examined the impact of pre existing diabetes on organ dysfunction during sepsis. Their main conclusion was that diabetes patients, relative to non-diabetics, were less likely to develop respiratory failure and more likely to develop renal failure during the course of sepsis. PMID- 19291262 TI - Extending international sepsis epidemiology: the impact of organ dysfunction. AB - In the previous issue of Critical Care, Blanco and colleagues contributed to a growing body of literature on the international epidemiology of severe sepsis. Taken together, these studies confirm that the sepsis incidence is high, that the development of organ dysfunction is a major determinant of mortality and that the occurrence of organ dysfunction is influenced by chronic comorbid medical conditions. It is clear that early detection of organ dysfunction and serial sequential organ dysfunction scoring provides us with the best chance to optimize clinical care. Identifying factors that contribute to the development of organ dysfunction in sepsis will lead to the development of new treatment modalities that will reduce mortality. Future studies must therefore focus on the impact of new treatment modalities for preventing progression to multiple organ dysfunction syndrome and consequent mortality in sepsis. PMID- 19291264 TI - Reality check for malaria proteomics. AB - New studies highlight the wide diversity of post-translational protein modifications in the intra-erythrocytic stages of the malaria parasite, raising new avenues for inquiry. PMID- 19291263 TI - Hypoxia. Regulation of NFkappaB signalling during inflammation: the role of hydroxylases. AB - NFkappaB is a master regulator of innate immunity and inflammatory signalling. Microenvironmental hypoxia has long been identified as being coincident with chronic inflammation. The contribution of microenvironmental hypoxia to NFkappaB induced inflammation has more recently been appreciated. Identification of the co regulation of NFkappaB and hypoxia inducible factor (HIF) pathways by 2-oxo glutarate-dependent hydroxylase family members has highlighted an intimate relationship between NFkappaB inflammatory signalling and HIF-mediated hypoxic signalling pathways. Adding another layer of complexity to our understanding of the role of NFkappaB inflammatory signalling by hypoxia is the recent recognition of the contribution of basal NFkappaB activity to HIF-1alpha transcription. This observation implicates an important and previously unappreciated role for NFkappaB in inflammatory disease where HIF-1alpha is activated. The present review will discuss recent literature pertaining to the regulation of NFkappaB inflammatory signalling by hypoxia and some of the inflammatory diseases where this may play an important role. Furthermore, we will discuss the potential for prolylhydroxylase inhibitors in inflammatory disease. PMID- 19291266 TI - In-flight medical emergencies: time for a registry? AB - When a passenger becomes sick while flying on board a commercial airline flight, the cabin crew commonly solicit the assistance of a volunteer physician. Although in-flight medical emergencies take place every day, little is known about the epidemiology of these events. A new study by Sand and colleagues sheds light on the incidence of specific illnesses that occurred on board commercial flights. PMID- 19291267 TI - Yeasts as models in evolutionary biology. AB - A report of the workshop 'Evolutionary and Environmental Genomics of Yeasts', Heidelberg, Germany, 1-5 October 2008. PMID- 19291268 TI - Innovative combination strategy to enhance effect and diminish adverse effects of glucocorticoids: another promise? AB - In a paper by Zimmermann and colleagues in this issue of Arthritis Research & Therapy, results of extended laboratory research with the drug combination of prednisolone and dipyridamole are reported. There seems to be a boost and extension of the glucocorticoid effect by the combination, without a clear increase of adverse effects, potentially allowing the application of lower dosages. However, laboratory models are not patients and the glucocorticoid mechanisms leading to effects and adverse effects are manifold. The next required step will be to demonstrate the improved therapeutic window in patients in adequate comparative clinical trials, assessing predefined beneficial effects and adverse effects in a standardized way. PMID- 19291269 TI - Recent developments in anti-rheumatic drugs in pediatrics: treatment of juvenile idiopathic arthritis. AB - Juvenile idiopathic arthritis (JIA) is the most common autoimmune autoinflammatory disease in childhood and affects approximately 1 in 1,000 children. Despite advances in diagnosis and treatment options, JIA remains a chronic condition for most affected children. Recent evidence suggests that disease control at onset may determine the tempo of subsequent disease course and long-term outcomes, and raises the concept of a therapeutic window of opportunity in patients with JIA. This underscores the importance of early aggressive treatment in patients with JIA. With the advent of novel biologic therapeutics, the repertoire of agents available for treatment of children with JIA has greatly increased. The present article will summarize recent developments in the medical treatment of children with JIA and will offer insights into emerging therapies. PMID- 19291270 TI - In ovo omnia: diversification by duplication in fish and other vertebrates. AB - Gene and genome duplications are considered to be the main evolutionary mechanisms contributing to the unrivalled biodiversity of bony fish. New studies of vitellogenin yolk proteins, including a report in BMC Evolutionary Biology, reveal that the genes underlying key evolutionary innovations and adaptations have undergone complex patterns of duplication and functional evolution. PMID- 19291271 TI - Patients with cancer on the ICU: the times they are changing. AB - A recent paper by Taccone and coworkers showed that 15% of patients from 198 European intensive care units (ICUs) had a malignancy, mostly solid tumors but also hematological malignancies. Over the past years, the prognosis of cancer patients has improved significantly, even when ICU admission is necessary. Refusal of ICU admission should not be based on a diagnosis of cancer as the underlying condition. In contrast, these decisions should be based on the availability of treatment options, and on patients' own preferences. PMID- 19291272 TI - Rac and Rho driving tumor invasion: who's at the wheel? AB - Genome-wide analysis of regulators of Rho-family small GTPases has identified critical elements that control the morphology and invasive behavior of melanoma cells. PMID- 19291274 TI - Are we training pit bulls to review our manuscripts? AB - Good early training of graduate students and postdocs is needed to prevent them turning into future generations of manuscript-savaging reviewers. How can we intercalate typical papers into our training? PMID- 19291273 TI - Transforming growth factor-beta in breast cancer: too much, too late. AB - The contribution of transforming growth factor (TGF)beta to breast cancer has been studied from a myriad perspectives since seminal studies more than two decades ago. Although the action of TGFbeta as a canonical tumor suppressor in breast is without a doubt, there is compelling evidence that TGFbeta is frequently subverted in a malignant plexus that drives breast cancer. New knowledge that TGFbeta regulates the DNA damage response, which underlies cancer therapy, reveals another facet of TGFbeta biology that impedes cancer control. Too much TGFbeta, too late in cancer progression is the fundamental motivation for pharmaceutical inhibition. PMID- 19291275 TI - Building bridges from 'omics' to cell biology. AB - A report from the Keystone Symposium on Molecular and Cellular Biology, 'Omics Meets Cell Biology', Breckenridge, Colorado, 25-30 January, 2009. PMID- 19291276 TI - The transition from ductal carcinoma in situ to invasive breast cancer: the other side of the coin. AB - The factors associated with the progression of ductal carcinoma in situ (DCIS) to invasive breast cancer are poorly understood. Many studies of this subject focus on the role of molecular and genetic alterations in the neoplastic epithelial cells. However, emerging evidence suggests that transition from DCIS to invasive cancer is strongly dependent upon alterations in the microenvironment. The potential roles of myoepithelial cells and of stromal-epithelial interaction are of particular interest in this regard. PMID- 19291277 TI - Gene expression changes with a 'non-injurious' ventilation strategy. PMID- 19291278 TI - Bench-to-bedside review: oxygen as a drug. AB - Oxygen is one of the most commonly used therapeutic agents. Injudicious use of oxygen at high partial pressures (hyperoxia) for unproven indications, its known toxic potential, and the acknowledged roles of reactive oxygen species in tissue injury led to skepticism regarding its use. A large body of data indicates that hyperoxia exerts an extensive profile of physiologic and pharmacologic effects that improve tissue oxygenation, exert anti-inflammatory and antibacterial effects, and augment tissue repair mechanisms. These data set the rationale for the use of hyperoxia in a list of clinical conditions characterized by tissue hypoxia, infection, and consequential impaired tissue repair. Data on regional hemodynamic effects of hyperoxia and recent compelling evidence on its anti inflammatory actions incited a surge of interest in the potential therapeutic effects of hyperoxia in myocardial revascularization and protection, in traumatic and nontraumatic ischemicanoxic brain insults, and in prevention of surgical site infections and in alleviation of septic and nonseptic local and systemic inflammatory responses. Although the margin of safety between effective and potentially toxic doses of oxygen is relatively narrow, the ability to carefully control its dose, meticulous adherence to currently accepted therapeutic protocols, and individually tailored treatment regimens make it a cost-effective safe drug. PMID- 19291279 TI - Clinical review: Ventilatory strategies for obstetric, brain-injured and obese patients. AB - The ventilatory management of patients with acute respiratory failure is supported by good evidence, aiming to reduce lung injury by pressure limitation and reducing the duration of ventilatory support by regular assessment for discontinuation. Certain patient groups, however, due to their altered physiology or disease-specific complications, may require some variation in usual ventilatory management. The present manuscript reviews the ventilatory management in three special populations, namely the patient with brain injury, the pregnant patient and the morbidly obese patient. PMID- 19291280 TI - Two devils: hypernatremia and hyponatremia can show faces to the same patient in the intensive care unit. PMID- 19291281 TI - Automics: an integrated platform for NMR-based metabonomics spectral processing and data analysis. AB - BACKGROUND: Spectral processing and post-experimental data analysis are the major tasks in NMR-based metabonomics studies. While there are commercial and free licensed software tools available to assist these tasks, researchers usually have to use multiple software packages for their studies because software packages generally focus on specific tasks. It would be beneficial to have a highly integrated platform, in which these tasks can be completed within one package. Moreover, with open source architecture, newly proposed algorithms or methods for spectral processing and data analysis can be implemented much more easily and accessed freely by the public. RESULTS: In this paper, we report an open source software tool, Automics, which is specifically designed for NMR-based metabonomics studies. Automics is a highly integrated platform that provides functions covering almost all the stages of NMR-based metabonomics studies. Automics provides high throughput automatic modules with most recently proposed algorithms and powerful manual modules for 1D NMR spectral processing. In addition to spectral processing functions, powerful features for data organization, data pre-processing, and data analysis have been implemented. Nine statistical methods can be applied to analyses including: feature selection (Fisher's criterion), data reduction (PCA, LDA, ULDA), unsupervised clustering (K Mean) and supervised regression and classification (PLS/PLS-DA, KNN, SIMCA, SVM). Moreover, Automics has a user-friendly graphical interface for visualizing NMR spectra and data analysis results. The functional ability of Automics is demonstrated with an analysis of a type 2 diabetes metabolic profile. CONCLUSION: Automics facilitates high throughput 1D NMR spectral processing and high dimensional data analysis for NMR-based metabonomics applications. Using Automics, users can complete spectral processing and data analysis within one software package in most cases. Moreover, with its open source architecture, interested researchers can further develop and extend this software based on the existing infrastructure. PMID- 19291282 TI - Multiple sequence-directed possibilities provide a pool of nucleosome position choices in different states of activity of a gene. AB - BACKGROUND: Genome-wide mappings of nucleosome occupancy in different species have shown presence of well-positioned nucleosomes. While the DNA sequences may help decide their locations, the observed positions in vivo are end-results of chromatin remodeling, the state of gene activity and binding of the sequence specific factors to the DNA, all of which influence nucleosome positions. Thus, the observed nucleosome locations in vivo do not reflect the true contribution of DNA sequence to the mapped position. Moreover, the naturally occurring nucleosome positioning sequences are known to guide multiple translational positionings. RESULTS: We show that yeast SNR6, a gene transcribed by RNA polymerase III, constitutes nucleosome-positioning sequence. In the absence of a chromatin remodeler or any factor binding, the gene sequence confers a unique rotational phase to nucleosomes in the gene region, and directs assembly of several translationally positioned nucleosomes on approximately 1.2 kb DNA from the gene locus, including the short approximately 250 bp gene region. Mapping of all these gene sequence-directed nucleosome positions revealed that the array of nucleosomes in the gene upstream region occupy the same positions as those observed in vivo but the nucleosomes on the gene region can be arranged in three distinct registers. Two of these arrangements differ from each other in the position of only one nucleosome, and match with the nucleosome positions on the gene in repressed and active states in vivo, where the gene-specific factor is known to occupy the gene in both the states. The two positions are interchanged by an ATP-dependent chromatin remodeler in vivo. The third register represents the positions which block the access of the factor to the gene promoter elements. CONCLUSION: On a gene locus, multiple nucleosome positions are directed by a gene sequence to provide a pool of possibilities, out of which the preferred ones are selected by the chromatin remodeler and transcription factor of the gene under different states of activity of the gene. PMID- 19291283 TI - A compact VEGF signature associated with distant metastases and poor outcomes. AB - BACKGROUND: Tumor metastases pose the greatest threat to a patient's survival, and thus, understanding the biology of disseminated cancer cells is critical for developing effective therapies. METHODS: Microarrays and immunohistochemistry were used to analyze primary breast tumors, regional (lymph node) metastases, and distant metastases in order to identify biological features associated with distant metastases. RESULTS: When compared with each other, primary tumors and regional metastases showed statistically indistinguishable gene expression patterns. Supervised analyses comparing patients with distant metastases versus primary tumors or regional metastases showed that the distant metastases were distinct and distinguished by the lack of expression of fibroblast/mesenchymal genes, and by the high expression of a 13-gene profile (that is, the 'vascular endothelial growth factor (VEGF) profile') that included VEGF, ANGPTL4, ADM and the monocarboxylic acid transporter SLC16A3. At least 8 out of 13 of these genes contained HIF1alpha binding sites, many are known to be HIF1alpha-regulated, and expression of the VEGF profile correlated with HIF1alpha IHC positivity. The VEGF profile also showed prognostic significance on tests of sets of patients with breast and lung cancer and glioblastomas, and was an independent predictor of outcomes in primary breast cancers when tested in models that contained other prognostic gene expression profiles and clinical variables. CONCLUSION: These data identify a compact in vivo hypoxia signature that tends to be present in distant metastasis samples, and which portends a poor outcome in multiple tumor types.This signature suggests that the response to hypoxia includes the ability to promote new blood and lymphatic vessel formation, and that the dual targeting of multiple cell types and pathways will be needed to prevent metastatic spread. PMID- 19291284 TI - Progress along developmental tracks for electronic health records implementation in the United States. AB - The development and implementation of electronic health records (EHR) have occurred slowly in the United States. To date, these approaches have, for the most part, followed four developmental tracks: (a) Enhancement of immunization registries and linkage with other health records to produce Child Health Profiles (CHP), (b) Regional Health Information Organization (RHIO) demonstration projects to link together patient medical records, (c) Insurance company projects linked to ICD-9 codes and patient records for cost-benefit assessments, and (d) Consortia of EHR developers collaborating to model systems requirements and standards for data linkage. Until recently, these separate efforts have been conducted in the very silos that they had intended to eliminate, and there is still considerable debate concerning health professionals access to as well as commitment to using EHR if these systems are provided. This paper will describe these four developmental tracks, patient rights and the legal environment for EHR, international comparisons, and future projections for EHR expansion across health networks in the United States. PMID- 19291285 TI - Dramatic response of a gastrointestinal stromal tumor to neadjuvant imatinib therapy. AB - Gastrointestinal stromal tumors (GISTs) are the most common sarcoma of the alimentary tract and are believed to derive from the interstitial Cell of Cajal. Imatinib mesylate (Gleevec; Novartis, Basel, Switzerland) has revolutionized the treatment of GISTs and is generally used in the metastatic and adjuvant settings. We report the case of a 61-year old man who was treated with neoadjuvant imatinib for a massive gastric GIST with the hope of avoiding a potential multi-visceral resection. PMID- 19291286 TI - The influence of solid/liquid separation techniques on the sugar yield in two step dilute acid hydrolysis of softwood followed by enzymatic hydrolysis. AB - BACKGROUND: Two-step dilute acid hydrolysis of softwood, either as a stand-alone process or as pretreatment before enzymatic hydrolysis, is considered to result in higher sugar yields than one-step acid hydrolysis. However, this requires removal of the liquid between the two steps. In an industrial process, filtration and washing of the material between the two steps is difficult, as it should be performed at high pressure to reduce energy demand. Moreover, the application of pressure leads to more compact solids, which may affect subsequent processing steps. This study was carried out to investigate the influence of pressing the biomass, in combination with the effects of not washing the material, on the sugar yield obtained from two-step dilute acid hydrolysis, with and without subsequent enzymatic digestion of the solids. RESULTS: Washing the material between the two acid hydrolysis steps, followed by enzymatic digestion, resulted in recovery of 96% of the mannose and 81% of the glucose (% of the theoretical) in the liquid fraction, regardless of the choice of dewatering method (pressing or vacuum filtration). Not washing the solids between the two acid hydrolysis steps led to elevated acidity of the remaining solids during the second hydrolysis step, which resulted in lower yields of mannose, 85% and 74% of the theoretical, for the pressed and vacuum-filtered slurry, respectively, due to sugar degradation. However, this increase in acidity resulted in a higher glucose yield (94.2%) from pressed slurry than from filtered slurry (77.6%). CONCLUSION: Pressing the washed material between the two acid hydrolysis steps had no significant negative effect on the sugar yields of the second acid hydrolysis step or on enzymatic hydrolysis. Not washing the material resulted in a harsher second acid hydrolysis step, which caused greater degradation of the sugars during subsequent acid hydrolysis of the solids, particularly in case of the vacuum-filtered solids. However, pressing in combination with not washing the material between the two steps enhanced the sugar yield of the enzymatic digestion step. Hence, it is suggested that the unwashed slurry be pressed to as high a dry matter content as possible between the two acid hydrolysis stages in order to achieve high final sugar yields. PMID- 19291287 TI - Anopheles pseudowillmori is the predominant malaria vector in Motuo County, Tibet Autonomous Region. AB - BACKGROUND: Malaria is endemic in Linzhi Prefecture in the Tibet Autonomous Region (TAR), but the vector for malaria transmission had never been identified. METHODS: Adult Anopheles spp. were collected in Motuo County, Linzhi Prefecture on the Sino-Indian border in July and August, 2007. Multiplex PCR was adopted for species identification, and a nested PCR approach was used to detect sporozoites in the salivary glands of the mosquitoes. RESULTS: 3,675 mosquitoes of the Anopheles maculatus group were collected and processed for species identification. Among them, 3,602 (98.0%) were Anopheles pseudowillmori and 73 (2.0%) were Anopheles willmori. The Plasmodium vivax SSUrDNA fragment was amplified in two of 360 pooled An. pseudowillmori samples. CONCLUSION: The local An. maculatus group comprises the species An. pseudowillmori and An. willmori. Anopheles pseudowillmori is considered the sole malaria vector in Motuo County in Linzhi Prefecture. PMID- 19291288 TI - Nonradioactive heteroduplex tracking assay for the detection of minority-variant chloroquine-resistant Plasmodium falciparum in Madagascar. AB - BACKGROUND: Strains of Plasmodium falciparum genetically resistant to chloroquine (CQ) due to the presence of pfcrt 76T appear to have been recently introduced to the island of Madagascar. The prevalence of such resistant genotypes is reported to be low (< 3%) when evaluated by conventional PCR. However, these methods are insensitive to low levels of mutant parasites present in patients with polyclonal infections. Thus, the current estimates may be an under representation of the prevalence of the CQ-resistant P. falciparum isolates on the island. Previously, minority variant chloroquine resistant parasites were described in Malawian patients using an isotopic heteroduplex tracking assay (HTA), which can detect pfcrt 76T-bearing P. falciparum minority variants in individual patients that were undetectable by conventional PCR. However, as this assay required a radiolabeled probe, it could not be used in many resource-limited settings. METHODS: This study describes a digoxigenin (DIG)-labeled chemiluminescent heteroduplex tracking assay (DIG-HTA) to detect pfcrt 76T-bearing minority variant P. falciparum. This assay was compared to restriction fragment length polymorphism (RFLP) analysis and to the isotopic HTA for detection of genetically CQ-resistant parasites in clinical samples. RESULTS: Thirty one clinical P. falciparum isolates (15 primary isolates and 16 recurrent isolates) from 17 Malagasy children treated with CQ for uncomplicated malaria were genotyped for the pfcrt K76T mutation. Two (11.7%) of 17 patients harboured genetically CQ resistant P. falciparum strains after therapy as detected by HTA. RFLP analysis failed to detect any pfcrt K76T-bearing isolates. CONCLUSION: These findings indicate that genetically CQ-resistant P. falciparum are more common than previously thought in Madagascar even though the fitness of the minority variant pfcrt 76T parasites remains unclear. In addition, HTAs for malaria drug resistance alleles are promising tools for the surveillance of anti-malarial resistance. The use of a non-radioactive label allows for the use of HTAs in malaria endemic countries. PMID- 19291289 TI - Function of anterior talofibular and calcaneofibular ligaments during in-vivo motion of the ankle joint complex. AB - BACKGROUND: Despite the numerous in-vitro studies on the mechanical properties and simulated injury mechanisms of the anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL), the in-vivo biomechanical behavior of these two ligaments has not yet been described. METHODS: Apparent length of the ATFL and CFL was measured in four ankles in healthy male subjects between 32 and 45 years of age (two left and two right) during a dorsiflexion-plantarflexion and supination-pronation arc of motion using a combined dual-orthogonal fluoroscopic and magnetic resonance imaging technique. RESULTS: The ATFL elongated from the neutral position at 16.3 +/- 3.0 mm to 20.8 +/- 2.7 mm at maximal plantarflexion and shortened significantly from the neutral position to 13.9 +/- 2.9 mm at maximal dorsiflexion (p = 0.01). The CFL shortened from the neutral position at 28.0 +/- 2.9 mm to 26.6 +/- 2.2 mm at maximal plantarflexion (p = 0.08) and elongated significantly from the neutral position to 29.9 +/- 3.0 mm at maximal dorsiflexion (p = 0.003). The ATFL elongated significantly from 14.8 +/- 2.5 mm at maximal pronation to 17.4 +/- 3.0 mm at maximal supination (p = 0.08). At the same time, the CFL shortened from 31.0 +/- 3.8 mm at maximal pronation to 26.9 +/ 3.6 mm at maximal supination (p = 0.02). CONCLUSION: The results showed that the ATFL elongates more during plantarflexion and supination whereas the CFL increases in length with dorsiflexion and pronation. Concurrently, these data also demonstrated the reciprocal function between the two ligaments. While one shortens, the other one elongates. The different elongation of the ATFL and CFL during the same motion arc suggests that under excessive loading conditions the ATFL might be more vulnerable in plantarflexion and supination while the CFL might be more susceptible to injury in dorsiflexion and pronation. Furthermore, in the case of surgical reconstruction the grafts used to reconstruct the two ligaments may need to be tensioned at different positions of the ankle in order to reproduce their natural in vivo function. PMID- 19291290 TI - Local increase in trapezius muscle oxygenation during and after acupuncture. AB - PURPOSE: This study aimed to compare the trapezius muscle blood volume and oxygenation in the stimulation region and in a distant region in the same muscle during acupuncture stimulation (AS). We hypothesized that AS provokes a localized increase in muscle blood volume and oxygenation in the stimulation region. METHODS: Two sets of near-infrared spectrometer (NIRS) probes, with 40-mm light source detector spacing, were placed on the right trapezius muscle, with a 50-mm distance between the probes. Changes in muscle oxygenation (oxy-Hb) and blood volume (t-Hb) in stimulation and distant regions (50 mm away from the stimulation point) were measured using NIRS. Nine healthy acupuncture-experienced subjects were chosen as the experimental (AS) group, and 10 healthy acupuncture experienced subjects were chosen for the control (no AS) group. Measurements began with a 3-min rest period, followed by "Jakutaku" (AS) for 2 min, and recovery after stimulation. RESULTS: There was a significant increase in oxy-Hb (60.7 muM at maximum) and t-Hb (48.1 muM at maximum) in the stimulation region compared to the distant region. In the stimulation region, a significant increase in oxy-Hb and t-Hb compared with the pre-stimulation level was first noted at 58.5 s and 13.5 s, respectively, after the onset of stimulation. CONCLUSION: In conclusion, oxygenation and blood volume increased, indicating elevated blood flow to the small vessels, not in the distant region used in this study, but in the stimulation region of the trapezius muscle during and after a 2-min AS. PMID- 19291291 TI - RSCA genotyping of MHC for high-throughput evolutionary studies in the model organism three-spined stickleback Gasterosteus aculeatus. AB - BACKGROUND: In all jawed vertebrates, highly polymorphic genes of the major histocompatibility complex (MHC) encode antigen presenting molecules that play a key role in the adaptive immune response. Their polymorphism is composed of multiple copies of recently duplicated genes, each possessing many alleles within populations, as well as high nucleotide divergence between alleles of the same species. Experimental evidence is accumulating that MHC polymorphism is a result of balancing selection by parasites and pathogens. In order to describe MHC diversity and analyse the underlying mechanisms that maintain it, a reliable genotyping technique is required that is suitable for such highly variable genes. RESULTS: We present a genotyping protocol that uses Reference Strand-mediated Conformation Analysis (RSCA), optimised for recently duplicated MHC class IIB genes that are typical for many fish and bird species, including the three-spined stickleback, Gasterosteus aculeatus. In addition we use a comprehensive plasmid library of MHC class IIB alleles to determine the nucleotide sequence of alleles represented by RSCA allele peaks. Verification of the RSCA typing by cloning and sequencing demonstrates high congruency between both methods and provides new insight into the polymorphism of classical stickleback MHC genes. Analysis of the plasmid library additionally reveals the high resolution and reproducibility of the RSCA technique. CONCLUSION: This new RSCA genotyping protocol offers a fast, but sensitive and reliable way to determine the MHC allele repertoire of three spined sticklebacks. It therefore provides a valuable tool to employ this highly polymorphic and adaptive marker in future high-throughput studies of host parasite co-evolution and ecological speciation in this emerging model organism. PMID- 19291292 TI - Young adult obese subjects with and without insulin resistance: what is the role of chronic inflammation and how to weigh it non-invasively? AB - BACKGROUND: Obesity is a leading risk factor for metabolic syndrome whose further expression is non-alcoholic fatty liver disease. Metabolic syndrome is associated with a proinflammatory state that contributes to insulin resistance. Finally, a "metabolically benign obesity" that is not accompanied by insulin resistance has recently been postulated to exist. AIM: To find whether any inflammation markers were independently associated with the presence of insulin resistance, evaluating specific anthropometric, ultrasonographic and laboratory parameters in a population of young adult obese subjects. METHODS: Of forty two young individuals, divided into two groups (with or without insulin resistance), were studied serum C-reactive protein and fibrinogen as indexes of chronic pro inflammatory status. Body mass index, waist circumference and metabolic syndrome presence were assessed as part of the metabolic evaluation. Ultrasonography weighted visceral and subcutaneous abdominal fat thickness, spleen size as longitudinal diameter and liver hyperechogenicity. RESULTS AND DISCUSSION: Serum C-reactive protein and fibrinogen as well as spleen longitudinal diameter were significantly increased in the obese young with insulin resistance compared to non-insulin resistance group. Insulin resistance was significantly associated with hepatic steatosis score at sonography (r = 0.33, P = 0.03), spleen longitudinal diameter (r = 0.35, P = 0.02) and C-reactive protein (r = 0.38, P = 0.01), but not with body mass index, visceral or subcutaneous abdominal adipose tissue, waist circumference and fibrinogen (P = 0.18, 0.46, 0.33, 0.37 and 0.4, respectively). Steatosis score at sonography was well associated with spleen volume (rho = 0.40, P = 0.01) and C-reactive protein levels (rho = 0.49, P = 0.002). Metabolic syndrome was much more frequent in obese patients with insulin resistance. These findings show that in young adults the only abdominal adiposity without insulin resistance, plays a scarce role in determining hepatic steatosis as well as metabolic syndrome. CONCLUSION: Increases in spleen size and CRP levels represent a reliable tool in diagnosing insulin resistance. PMID- 19291293 TI - Validating an alternate version of the chewing function questionnaire in partially dentate patients. AB - BACKGROUND: The aim of this study was to investigate the dimensionality, reliability, and validity of an alternate version of the chewing function questionnaire in partially dentate patients in Japan. METHODS: Subjects were partially dentate patients who attended the prosthodontic clinic at Tokyo Medical and Dental University (N = 491, 71% women, mean age (+/- SD): 63.0 +/- 11.5 years). The questionnaire asked each subject to rate his or her ability to chew 20 common Japanese foods. For each individual, responses were combined to yield a chewing function summary score, with higher scores indicating better self reported chewing ability. We used exploratory factor analysis to investigate the scores' dimensionality. For validity assessment, we computed the correlations between the chewing function score and oral health-related quality of life (OHRQoL, as measured by the Japanese 14-item Oral Health Impact Profile (OHIP 14)) Internal consistency of scores and test-retest reliability were investigated by asking a subset of subjects (N = 62) to complete the questionnaire twice, 2 weeks apart. RESULTS: Exploratory factor analysis provided some evidence that self-reported chewing ability can be characterized by a summary score as the original authors suggest. Support for the validity of chewing function scores using the alternate version of the questionnaire was derived from correlations with OHIP-14 scores (r = -0.46, 95% confidence interval (CI): -0.53 to -0.39); thus, better chewing ability was associated with less impaired OHRQoL. Internal consistency was 'satisfactory,' with a Cronbach's alpha of 0.90 (lower limit of 95% CI: 0.89). The test-retest reliability was 'good,' with an intraclass correlation coefficient of 0.69 (95% CI: 0.56 to 0.82). CONCLUSION: The alternate version of the chewing function questionnaire can be used as a stand-alone instrument because of the demonstrated reliability and validity of scores obtained using the questionnaire in partially dentate patients. PMID- 19291294 TI - APRIL is overexpressed in cancer: link with tumor progression. AB - BACKGROUND: BAFF and APRIL share two receptors - TACI and BCMA - and BAFF binds to a third receptor, BAFF-R. Increased expression of BAFF and APRIL is noted in hematological malignancies. BAFF and APRIL are essential for the survival of normal and malignant B lymphocytes, and altered expression of BAFF or APRIL or of their receptors (BCMA, TACI, or BAFF-R) have been reported in various B-cell malignancies including B-cell non-Hodgkin's lymphoma, chronic lymphocytic leukemia, Hodgkin's lymphoma, multiple myeloma, and Waldenstrom's macroglobulinemia. METHODS: We compared the expression of BAFF, APRIL, TACI and BAFF-R gene expression in 40 human tumor types - brain, epithelial, lymphoid, germ cells - to that of their normal tissue counterparts using publicly available gene expression data, including the Oncomine Cancer Microarray database. RESULTS: We found significant overexpression of TACI in multiple myeloma and thyroid carcinoma and an association between TACI expression and prognosis in lymphoma. Furthermore, BAFF and APRIL are overexpressed in many cancers and we show that APRIL expression is associated with tumor progression. We also found overexpression of at least one proteoglycan with heparan sulfate chains (HS), which are coreceptors for APRIL and TACI, in tumors where APRIL is either overexpressed or is a prognostic factor. APRIL could induce survival or proliferation directly through HS proteoglycans. CONCLUSION: Taken together, these data suggest that APRIL is a potential prognostic factor for a large array of malignancies. PMID- 19291295 TI - Kerfdr: a semi-parametric kernel-based approach to local false discovery rate estimation. AB - BACKGROUND: The use of current high-throughput genetic, genomic and post-genomic data leads to the simultaneous evaluation of a large number of statistical hypothesis and, at the same time, to the multiple-testing problem. As an alternative to the too conservative Family-Wise Error-Rate (FWER), the False Discovery Rate (FDR) has appeared for the last ten years as more appropriate to handle this problem. However one drawback of FDR is related to a given rejection region for the considered statistics, attributing the same value to those that are close to the boundary and those that are not. As a result, the local FDR has been recently proposed to quantify the specific probability for a given null hypothesis to be true. RESULTS: In this context we present a semi-parametric approach based on kernel estimators which is applied to different high-throughput biological data such as patterns in DNA sequences, genes expression and genome wide association studies. CONCLUSION: The proposed method has the practical advantages, over existing approaches, to consider complex heterogeneities in the alternative hypothesis, to take into account prior information (from an expert judgment or previous studies) by allowing a semi-supervised mode, and to deal with truncated distributions such as those obtained in Monte-Carlo simulations. This method has been implemented and is available through the R package kerfdr via the CRAN or at (http://stat.genopole.cnrs.fr/software/kerfdr). PMID- 19291296 TI - Whole genome surveys of rice, maize and sorghum reveal multiple horizontal transfers of the LTR-retrotransposon Route66 in Poaceae. AB - BACKGROUND: Horizontal transfers (HTs) refer to the transmission of genetic material between phylogenetically distant species. Although most of the cases of HTs described so far concern genes, there is increasing evidence that some involve transposable elements (TEs) in Eukaryotes. The availability of the full genome sequence of two cereal species, (i.e. rice and Sorghum), as well as the partial genome sequence of maize, provides the opportunity to carry out genome wide searches for TE-HTs in Poaceae. RESULTS: We have identified an LTR retrotransposon, that we named Route66, with more than 95% sequence identity between rice and Sorghum. Using a combination of in silico and molecular approaches, we are able to present a substantial phylogenetic evidence that Route66 has been transferred horizontally between Panicoideae and several species of the genus Oryza. In addition, we show that it has remained active after these transfers. CONCLUSION: This study constitutes a new case of HTs for an LTR retrotransposon and we strongly believe that this mechanism could play a major role in the life cycle of transposable elements. We therefore propose to integrate classe I elements into the previous model of transposable element evolution through horizontal transfers. PMID- 19291297 TI - Discovery and identification of potential biomarkers of pediatric acute lymphoblastic leukemia. AB - BACKGROUND: Acute lymphoblastic leukemia (ALL) is a common form of cancer in children. Currently, bone marrow biopsy is used for diagnosis. Noninvasive biomarkers for the early diagnosis of pediatric ALL are urgently needed. The aim of this study was to discover potential protein biomarkers for pediatric ALL. METHODS: Ninety-four pediatric ALL patients and 84 controls were randomly divided into a "training" set (45 ALL patients, 34 healthy controls) and a test set (49 ALL patients, 30 healthy controls and 30 pediatric acute myeloid leukemia (AML) patients). Serum proteomic profiles were measured using surface-enhanced laser desorption/ionization-time-of-flight mass spectroscopy (SELDI-TOF-MS). A classification model was established by Biomarker Pattern Software (BPS). Candidate protein biomarkers were purified by HPLC, identified by LC-MS/MS and validated using ProteinChip immunoassays. RESULTS: A total of 7 protein peaks (9290 m/z, 7769 m/z, 15110 m/z, 7564 m/z, 4469 m/z, 8937 m/z, 8137 m/z) were found with differential expression levels in the sera of pediatric ALL patients and controls using SELDI-TOF-MS and then analyzed by BPS to construct a classification model in the "training" set. The sensitivity and specificity of the model were found to be 91.8%, and 90.0%, respectively, in the test set. Two candidate protein peaks (7769 and 9290 m/z) were found to be down-regulated in ALL patients, where these were identified as platelet factor 4 (PF4) and pro platelet basic protein precursor (PBP). Two other candidate protein peaks (8137 and 8937 m/z) were found up-regulated in the sera of ALL patients, and these were identified as fragments of the complement component 3a (C3a). CONCLUSION: Platelet factor (PF4), connective tissue activating peptide III (CTAP-III) and two fragments of C3a may be potential protein biomarkers of pediatric ALL and used to distinguish pediatric ALL patients from healthy controls and pediatric AML patients. Further studies with additional populations or using pre-diagnostic sera are needed to confirm the importance of these findings as diagnostic markers of pediatric ALL. PMID- 19291298 TI - Detecting imipenem resistance in Acinetobacter baumannii by automated systems (BD Phoenix, Microscan WalkAway, Vitek 2); high error rates with Microscan WalkAway. AB - BACKGROUND: Increasing reports of carbapenem resistant Acinetobacter baumannii infections are of serious concern. Reliable susceptibility testing results remains a critical issue for the clinical outcome. Automated systems are increasingly used for species identification and susceptibility testing. This study was organized to evaluate the accuracies of three widely used automated susceptibility testing methods for testing the imipenem susceptibilities of A. baumannii isolates, by comparing to the validated test methods. METHODS: Selected 112 clinical isolates of A. baumanii collected between January 2003 and May 2006 were tested to confirm imipenem susceptibility results. Strains were tested against imipenem by the reference broth microdilution (BMD), disk diffusion (DD), Etest, BD Phoenix, MicroScan WalkAway and Vitek 2 automated systems. Data were analysed by comparing the results from each test method to those produced by the reference BMD test. RESULTS: MicroScan performed true identification of all A. baumannii strains while Vitek 2 unidentified one strain, Phoenix unidentified two strains and misidentified two strains. Eighty seven of the strains (78%) were resistant to imipenem by BMD. Etest, Vitek 2 and BD Phoenix produced acceptable error rates when tested against imipenem. Etest showed the best performance with only two minor errors (1.8%). Vitek 2 produced eight minor errors(7.2%). BD Phoenix produced three major errors (2.8%). DD produced two very major errors (1.8%) (slightly higher (0.3%) than the acceptable limit) and three major errors (2.7%). MicroScan showed the worst performance in susceptibility testing with unacceptable error rates; 28 very major (25%) and 50 minor errors (44.6%). CONCLUSION: Reporting errors for A. baumannii against imipenem do exist in susceptibility testing systems. We suggest clinical laboratories using MicroScan system for routine use should consider using a second, independent antimicrobial susceptibility testing method to validate imipenem susceptibility. Etest, whereever available, may be used as an easy method to confirm imipenem susceptibility. PMID- 19291299 TI - A novel method for screening the glutathione transferase inhibitors. AB - BACKGROUND: Glutathione transferases (GSTs) belong to the family of Phase II detoxification enzymes. GSTs catalyze the conjugation of glutathione to different endogenous and exogenous electrophilic compounds. Over-expression of GSTs was demonstrated in a number of different human cancer cells. It has been found that the resistance to many anticancer chemotherapeutics is directly correlated with the over-expression of GSTs. Therefore, it appears to be important to find new GST inhibitors to prevent the resistance of cells to anticancer drugs. In order to search for glutathione transferase (GST) inhibitors, a novel method was designed. RESULTS: Our results showed that two fragments of GST, named F1 peptide (GYWKIKGLV) and F2 peptide (KWRNKKFELGLEFPNL), can significantly inhibit the GST activity. When these two fragments were compared with several known potent GST inhibitors, the order of inhibition efficiency (measured in reactions with 2,4 dinitrochlorobenzene (CDNB) and glutathione as substrates) was determined as follows: tannic acid > cibacron blue > F2 peptide > hematin > F1 peptide > ethacrynic acid. Moreover, the F1 peptide appeared to be a noncompetitive inhibitor of the GST-catalyzed reaction, while the F2 peptide was determined as a competitive inhibitor of this reaction. CONCLUSION: It appears that the F2 peptide can be used as a new potent specific GST inhibitor. It is proposed that the novel method, described in this report, might be useful for screening the inhibitors of not only GST but also other enzymes. PMID- 19291300 TI - Marked increase of procalcitonin after the administration of anti-thymocyte globulin in patients before hematopoietic stem cell transplantation does not indicate sepsis: a prospective study. AB - INTRODUCTION: Procalcitonin (PCT) and C-reactive protein (CRP) are established markers of infection in the general population. In contrast, several studies reported falsely increased PCT levels in patients receiving T-cell antibodies. We evaluated the validity of these markers in patients scheduled for hemopoietic stem cell transplantation receiving anti-thymocyte globulin (ATG) during conditioning. We also assessed renal and liver functions and their relationship to PCT and CRP changes. METHODS: Twenty-six patients without clinical signs of infection were prospectively studied. ATG was administered in up to three doses over the course of 5 days. PCT, CRP, white blood cell (WBC) count, urea, creatinine, glomerular filtration rate, bilirubin, alanin amino-transferase (ALT), and gamma-glutamyl transferase (GGT) were assessed daily during ATG administration. Pharyngeal, nose, and rectal swabs and urine samples were cultured twice weekly. Blood cultures were obtained if clinical symptoms of infection were present. RESULTS: Baseline (BL) levels of both PCT and CRP before ATG administration were normal. WBC count decreased after ATG administration (P = 0.005). One day after ATG administration, both PCT and CRP levels increased significantly, returning to BL levels on day 4. Microbiological results were clinically unremarkable. There was no interrelationship between PCT levels and BL markers of renal or liver functions (P > 0.05 for all comparisons). Bilirubin and GGT were increased on days 2 to 5 and ALT was increased on day 3 (P < 0.05 versus BL). No difference in renal functions was observed. Three patients developed bacterial infection on days 7 to 11 with different dynamics of PCT and CRP. There was no association between the number of ATG doses and PCT levels or between the risk of developing infection and previous PCT levels. CONCLUSIONS: ATG triggered a marked early surge in PCT and CRP followed by a steady decrease over the course of 3 days. The dynamics of both PCT and CRP were similar and were not associated with infection. PCT levels were independent of renal and liver functions and were not predictive of further infectious complications. A direct effect of ATG on T lymphocytes could be the underlying mechanism. Hepatotoxic effect could be a contributing factor. Neither PCT nor CRP is a useful marker that can identify infection in patients receiving ATG. PMID- 19291301 TI - The efficacy and safety of a new fixed-dose combination of amodiaquine and artesunate in young African children with acute uncomplicated Plasmodium falciparum. AB - BACKGROUND: Artesunate (AS) plus amodiaquine (AQ) is one artemisinin-based combination (ACT) recommended by the WHO for treating Plasmodium falciparum malaria. Fixed-dose AS/AQ is new, but its safety and efficacy are hitherto untested. METHODS: A randomized, open-label trial was conducted comparing the efficacy (non-inferiority design) and safety of fixed (F) dose AS (25 mg)/AQ (67.5 mg) to loose (L) AS (50 mg) + AQ (153 mg) in 750, P. falciparum-infected children from Burkina Faso aged 6 months to 5 years. Dosing was by age. Primary efficacy endpoint was Day (D) 28, PCR-corrected, parasitological cure rate. Recipients of rescue treatment were counted as failures and new infections as cured. Documented, common toxicity criteria (CTC) graded adverse events (AEs) defined safety. RESULTS: Recruited and evaluable children numbered 750 (375/arm) and 682 (90.9%), respectively. There were 8 (AS/AQ) and 6 (AS+AQ) early treatment failures and one D7 failure (AS+AQ). Sixteen (AS/AQ) and 12 (AS+AQ) patients had recurrent parasitaemia (PCR new infections 10 and 6, respectively). Fourteen patients per arm required rescue treatment for vomiting/spitting out study drugs. Efficacy rates were 92.1% in both arms: AS/AQ = 315/342 (95% CI: 88.7-94.7) vs. AS+AQ = 313/340 (95% CI: 88.6-94.7). Non-inferiority was demonstrated at two sided alpha = 0.05: Delta (AS+AQ - AS/AQ) = 0.0% (95% CI: -4.1% to 4.0%). D28, Kaplan Meier PCR-corrected cure rates (all randomized children) were similar: 93.7% (AS/AQ) vs. 93.2% (AS+AQ) Delta = -0.5 (95% CI -4.2 to 3.0%). By D2, both arms had rapid parasite (F & L, 97.8% aparasitaemic) and fever (97.2% [F], 96.0% [L] afebrile) clearances.Both treatments were well tolerated. Drug-induced vomiting numbered 8/375 (2.1%) and 6/375 (1.6%) in the fixed and loose arms, respectively (p = 0.59). One patient developed asymptomatic, CTC grade 4 hepatitis (AST 1052, ALT 936). Technical difficulties precluded the assessment and risk of neutropaenia for all patients. CONCLUSION: Fixed dose AS/AQ was efficacious and well tolerated. These data support the use of this new fixed dose combination for treating P. falciparum malaria with continued safety monitoring. TRIAL REGISTRATION: Current Controlled Trials ISRCTN07576538. PMID- 19291302 TI - Characterization of NF-kB-mediated inhibition of catechol-O-methyltransferase. AB - BACKGROUND: Catechol-O-methyltransferase (COMT), an enzyme that metabolizes catecholamines, has recently been implicated in the modulation of pain. Specifically, low COMT activity is associated with heightened pain perception and development of musculoskeletal pain in humans as well as increased experimental pain sensitivity in rodents. RESULTS: We report that the proinflammatory cytokine tumor necrosis factor alpha (TNFalpha) downregulates COMT mRNA and protein in astrocytes. Examination of the distal COMT promoter (P2-COMT) reveals a putative binding site for nuclear factor kappaB (NF-kappaB), the pivotal regulator of inflammation and the target of TNFalpha. Cell culture assays and functional deletion analyses of the cloned P2-COMT promoter demonstrate that TNFalpha inhibits P2-COMT activity in astrocytes by inducing NF-kappaB complex recruitment to the specific kappaB binding site. CONCLUSION: Collectively, our findings provide the first evidence for NF-kappaB-mediated inhibition of COMT expression in the central nervous system, suggesting that COMT contributes to the pathogenesis of inflammatory pain states. PMID- 19291303 TI - Long term survivors with metastatic pancreatic adenocarcinoma treated with gemcitabine: a retrospective analysis. AB - BACKGROUND: Metastatic pancreatic adenocarcinoma has a short median overall survival (OS) of 5-6 months. However, a subgroup of patients survives more than 1 year. We analyzed the survival outcomes of this subgroup and evaluated clinical and pathological factors that might affect survival durations. METHODS: We identified 20 patients with metastatic or recurrent pancreatic adenocarcinoma who received single-agent gemcitabine and had an OS longer than 1 year. Baseline data available after the diagnosis of metastatic or recurrent disease was categorized as: 1) clinical/demographic data (age, gender, ECOG PS, number and location of metastatic sites); 2) Laboratory data (Hematocrit, hemoglobin, glucose, LDH, renal and liver function and CA19-9); 3) Pathologic data (margins, nodal status and grade); 4) Outcomes data (OS, Time to Treatment Failure (TTF), and 2 year OS). The lowest CA19-9 levels during treatment with gemcitabine were also recorded. We performed a univariate analysis with OS as the outcome variable. RESULTS: Baseline logarithm of CA19-9 and total bilirubin had a significant impact on OS (HR = 1.32 and 1.31, respectively). Median OS and TTF on gemcitabine were 26.9 (95% CI = 18 to 32) and 11.5 (95% CI = 9.0 to 14.3) months, respectively. Two-year OS was 56.4%, with 7 patients alive at the time of analysis. CONCLUSION: A subgroup of patients with metastatic pancreatic cancer has prolonged survival after treatment with gemcitabine. Only bilirubin and CA 19 9 levels were predictive of longer survival in this population. Further analysis of potential prognostic and predictive markers of response to treatment and survival are needed. PMID- 19291304 TI - Microparticle-induced release of B-lymphocyte regulators by rheumatoid synoviocytes. AB - INTRODUCTION: In the present study, we investigated the ability of microparticles isolated from synovial fluids from patients with rheumatoid arthritis or osteoarthritis to induce the synthesis and release of key cytokines of B lymphocyte modulation such as B cell-activating factor, thymic stroma lymphopoietin, and secretory leukocyte protease inhibitor by rheumatoid fibroblast-like synoviocytes. METHODS: Microparticles were analyzed in synovial fluids from patients with rheumatoid arthritis, osteoarthritis, microcristalline arthritis, and reactive arthritis. In addition, microparticle release after activation from various cell lines (CEM lymphocyte and THP-1 cells) was assessed. Microparticles were isolated by differential centrifugation, and quantitative determinations were carried out by prothrombinase assay after capture on immobilized annexin V. B cell-activating factor, thymic stroma lymphopoietin, and secretory leukocyte protease inhibitor release was evaluated by enzyme-linked immunosorbent assay. RESULTS: Microparticles isolated from synovial fluids obtained from rheumatoid arthritis and osteoarthritis patients or microparticles derived from activated THP-1 cells were able to induce B cell-activating factor, thymic stroma lymphopoietin, and secretory leukocyte protease inhibitor release by rheumatoid arthritis fibroblast-like synoviocytes. Conversely, CEM-lymphocytes derived microparticles generated by treatment with a combination of PHA, PMA and Adt-D did not promote the release of B cell-activating factor but favored the secretion of thymic stroma lymphopoietin and secretory leukocyte protease inhibitor by rheumatoid arthritis fibrobast-like synoviocytes. However, microparticles isolated from actinomycin D-treated CEM lymphocytes were not able to induce B cell-activating factor, thymic stroma lymphopoietin, or secretory leukocyte protease inhibitor release, indicating that microparticles derived from apoptotic T cells do not function as effectors in B-cell activation. CONCLUSIONS: These results demonstrate that microparticles are signalling structures that may act as specific conveyors in the triggered induction and amplification of autoimmunity. This study also indicates that microparticles have differential effects in the crosstalk between B lymphocytes and target cells of autoimmunity regarding the parental cells from which they derive. PMID- 19291305 TI - Arm rotated medially with supination - the ARMS variant: description of its surgical correction. AB - BACKGROUND: Patients who have suffered obstetric brachial plexus injury (OBPI) have a high incidence of musculoskeletal complications stemming from the initial nerve injury. The presence of muscle imbalances and contractures leads to typical bony changes affecting the shoulder, including the SHEAR (Scapular Hypoplasia, Elevation and Rotation) deformity. The SHEAR deformity commonly occurs in conjunction with Medial Rotation Contracture (MRC) of the arm. OBPI also causes muscle imbalances at the level of the forearm, that lead to a fixed supination deformity (SD) in a small number of patients. Both MRC and SD will cause severe functional limitations without surgical intervention. METHODS: Fourteen OBPI patients were diagnosed with MRC of the shoulder and SD of the forearm along with SHEAR deformity during a 16 month study period, with eight patients available to long-term follow-up (age range 2.2 - 18 years). Surgical correction of the MRC was performed as a triangle tilt or humeral osteotomy depending on the age of the child, after which, the patients were treated with a radial osteotomy to correct the fixed supination deformity. Function was assessed using the modified Mallet scale, examination of apparent supination and appearance of the extremity at rest. RESULTS: Significant functional improvements were observed in patients with surgical reconstruction. Mallet score increased by an average of 5.2 (p < 0.05). Overall forearm position was not significantly changed from an average of 5 degrees to an average of 34 degrees maximum apparent supination after both shoulder rotation and forearm rotation corrective surgeries. CONCLUSION: The simultaneous presence of two opposing deformities in the same limb will visually offset each other at the level of the wrist and hand, giving the false impression of neutral positioning of the limb. In reality, the neutral-appearing position of the hand indicates a fixed supination posture of the forearm in the face of a medial rotation contracture of the shoulder. Both of these deformities require surgical attention, and the presence of concurrent MRC and SD should be monitored for in OBPI patients. PMID- 19291306 TI - Population-specific gene expression in the plant pathogenic nematode Heterodera glycines exists prior to infection and during the onset of a resistant or susceptible reaction in the roots of the Glycine max genotype Peking. AB - BACKGROUND: A single Glycine max (soybean) genotype (Peking) reacts differently to two different populations of Heterodera glycines (soybean cyst nematode) within the first twelve hours of infection during resistant (R) and susceptible (S) reactions. This suggested that H. glycines has population-specific gene expression signatures. A microarray analysis of 7539 probe sets representing 7431 transcripts on the Affymetrix soybean GeneChip were used to identify population specific gene expression signatures in pre-infective second stage larva (pi-L2) prior to their infection of Peking. Other analyses focused on the infective L2 at 12 hours post infection (i-L2(12h)), and the infective sedentary stages at 3 days post infection (i-L2(3d)) and 8 days post infection (i-L2/L3(8d)). RESULTS: Differential expression and false discovery rate (FDR) analyses comparing populations of pi-L2 (i.e., incompatible population, NL1-RHg to compatible population, TN8) identified 71 genes that were induced in NL1-RHg as compared to TN8. These genes included putative gland protein G23G12, putative esophageal gland protein Hgg-20 and arginine kinase. The comparative analysis of pi-L2 identified 44 genes that were suppressed in NL1-RHg as compared to TN8. These genes included a different Hgg-20 gene, an EXPB1 protein and a cuticular collagen. By 12 h, there were 7 induced genes and 0 suppressed genes in NL1-RHg. By 3d, there were 9 induced and 10 suppressed genes in NL1-RHg. Substantial changes in gene expression became evident subsequently. At 8d there were 13 induced genes in NL1-RHg. This included putative gland protein G20E03, ubiquitin extension protein, putative gland protein G30C02 and beta-1,4 endoglucanase. However, 1668 genes were found to be suppressed in NL1-RHg. These genes included steroid alpha reductase, serine proteinase and a collagen protein. CONCLUSION: These analyses identify a genetic expression signature for these two populations both prior to and subsequently as they undergo an R or S reaction. The identification of genes like steroid alpha reductase and serine proteinase that are involved in feeding and nutritional uptake as being highly suppressed during the R response at 8d may indicate genes that the plant is targeting. The analyses also identified numerous putative parasitism genes that are differentially expressed. The 1668 genes that are suppressed in NL1-RHg, and hence induced in TN8 may represent genes that are important during the parasitic stages of H. glycines development. The potential for different arrays of putative parasitism genes to be expressed in different nematode populations may indicate how H. glycines evolve mechanisms to overcome resistance. PMID- 19291307 TI - Elevated P75NTR expression causes death of engrailed-deficient midbrain dopaminergic neurons by Erk1/2 suppression. AB - BACKGROUND: The homeodomain transcription factors Engrailed-1 and Engrailed-2 are required for the survival of mesencephalic dopaminergic (mesDA) neurons in a cell autonomous and gene-dose-dependent manner. Homozygote mutant mice, deficient of both genes (En1-/-;En2-/-), die at birth and exhibit a loss of all mesDA neurons by mid-gestation. In heterozygote animals (En1+/-;En2-/-), which are viable and fertile, postnatal maintenance of the nigrostriatal dopaminergic system is afflicted, leading to a progressive degeneration specific to this subpopulation and Parkinson's disease-like molecular and behavioral deficits. RESULTS: In this work, we show that the dose of Engrailed is inversely correlated to the expression level of the pan-neurotrophin receptor gene P75NTR (Ngfr). Loss of mesDA neurons in the Engrailed-null mutant embryos is caused by elevated expression of this neurotrophin receptor: Unusually, in this case, the cell death signal of P75NTR is mediated by suppression of Erk1/2 (extracellular-signal regulated kinase 1/2) activity. The reduction in expression of Engrailed, possibly related to the higher levels of P75NTR, also decreases mitochondrial stability. In particular, the dose of Engrailed determines the sensitivity to cell death induced by the classic Parkinson-model toxin MPTP and to inhibition of the anti-apoptotic members of the Bcl-2 family of proteins. CONCLUSION: Our study links the survival function of the Engrailed genes in developing mesDA neurons to the regulation of P75NTR and the sensitivity of these neurons to mitochondrial insult. The similarities to the disease etiology in combination with the nigral phenotype of En1+/-;En2-/- mice suggests that haplotype variations in the Engrailed genes and/or P75NTR that alter their expression levels could, in part, determine susceptibility to Parkinson's disease. PMID- 19291308 TI - Implications of the structure of human uridine phosphorylase 1 on the development of novel inhibitors for improving the therapeutic window of fluoropyrimidine chemotherapy. AB - BACKGROUND: Uridine phosphorylase (UPP) is a key enzyme of pyrimidine salvage pathways, catalyzing the reversible phosphorolysis of ribosides of uracil to nucleobases and ribose 1-phosphate. It is also a critical enzyme in the activation of pyrimidine-based chemotherapeutic compounds such a 5-fluorouracil (5-FU) and its prodrug capecitabine. Additionally, an elevated level of this enzyme in certain tumours is believed to contribute to the selectivity of such drugs. However, the clinical effectiveness of these fluoropyrimidine antimetabolites is hampered by their toxicity to normal tissue. In response to this limitation, specific inhibitors of UPP, such as 5-benzylacyclouridine (BAU), have been developed and investigated for their ability to modulate the cytotoxic side effects of 5-FU and its derivatives, so as to increase the therapeutic index of these agents. RESULTS: In this report we present the high resolution structures of human uridine phosphorylase 1 (hUPP1) in ligand-free and BAU inhibited conformations. The structures confirm the unexpected solution observation that the human enzyme is dimeric in contrast to the hexameric assembly present in microbial UPPs. They also reveal in detail the mechanism by which BAU engages the active site of the protein and subsequently disables the enzyme by locking the protein in a closed conformation. The observed inter-domain motion of the dimeric human enzyme is much greater than that seen in previous UPP structures and may result from the simpler oligomeric organization. CONCLUSION: The structural details underlying hUPP1's active site and additional surfaces beyond these catalytic residues, which coordinate binding of BAU and other acyclouridine analogues, suggest avenues for future design of more potent inhibitors of this enzyme. Notably, the loop forming the back wall of the substrate binding pocket is conformationally different and substantially less flexible in hUPP1 than in previously studied microbial homologues. These distinctions can be utilized to discover novel inhibitory compounds specifically optimized for efficacy against the human enzyme as a step toward the development of more effective chemotherapeutic regimens that can selectively protect normal tissues with inherently lower UPP activity. PMID- 19291309 TI - The roll-your-own cigarette market in Canada: a cross-sectional exploratory study. AB - BACKGROUND: Even though the use and prevalence of roll-your-own cigarettes (RYO) has been declining over the past decades, RYO remains important. Given the paucity of research examining RYO use, there is a need to better understand the current and potential future context of RYO use. METHODS: Data from the 2002 Canadian Tobacco Use Monitoring Survey (CTUMS) were used to examine RYO tobacco use among 23,341 Canadians aged 15 and older. Logistic regression models were conducted to examine factors which differentiate smokers who smoke RYO tobacco all of the time, most of the time or sometimes from smokers who do not smoke RYO tobacco. RESULTS: We found that 17% (n = 925,000) of current smokers in Canada reported smoking RYO. When compared to manufactured cigarette (MC) smokers, RYO users were heavier smokers, more addicted to nicotine, and less likely to consider quitting smoking. Lower income smokers were more likely to smoke RYO tobacco compared to smokers with high income. Conversely, smokers who had completed secondary school or university were less likely to smoke RYO tobacco compared to smokers who had not completed secondary school. CONCLUSION: This study demonstrates that RYO tobacco use is not a negligible problem within Canada and provides valuable new insight for developing future tobacco control initiatives for this population of smokers. PMID- 19291310 TI - Preferential uptake of the non steroid anti-inflammatory drug diclofenac into inflamed tissues after a single oral dose in rats. AB - BACKGROUND: Diclofenac is a nonsteroidal anti-inflammatory drug which is available as prescription (RX) and over-the-counter (OTC) medication for the systemic and topical treatment of painful and inflammatory conditions such as arthritis and back pain. This study was undertaken to investigate the distribution and retention of diclofenac and/or its metabolites in inflamed tissues, using the carrageenan-induced inflammation model and quantitative whole body autoradiography in rats. METHODS: [14C]diclofenac sodium was administrated as a single 2 mg/kg oral dose 1 h after injection of carrageenan into one front and one hind footpads and subcutaneously into the dorsum of the neck of rats. A control animal received saline injections. Three carrageenan-treated rats and one control rat were sacrificed at 1, 4, 8, and 24 h after [14C]diclofenac sodium administration (total of 4 rats/time point). The carcasses were immediately snap frozen and prepared for cryosectioning. Lengthwise whole-body sections (40 microm thick), including all major tissues, were obtained from different levels across the body. The tissue concentrations of total radiolabeled components were determined using quantitative autoradioluminography. RESULTS: The radioactivity patterns demonstrated that diclofenac and/or its metabolites preferentially distributed into the inflamed tissues. In unharmed tissues the distribution was similar in control and treated animals. The exposure, based on the areas under the tissue concentration versus time (AUC(0-tlast)), was 26 and 53 fold higher in the inflamed neck and inflamed footpads of treated animals than in control rats; the exposures in unharmed tissues were similar in the treated and control rats, and the AUC(0-tlast) was 17 fold higher in the inflamed paws than in the non inflamed footpads of the carrageenan-treated rats. The higher exposure in the inflamed tissues may be explained partly to the fact that the elimination of total radiolabeled components from inflamed tissues (t(1/2) = 6 h) appeared lower than from the corresponding unharmed tissues (t(1/2) = 2 h). CONCLUSION: This animal study demonstrated that diclofenac and/or its metabolites were rapidly and preferentially taken up and retained in inflamed tissues. Although there were theoretical considerations that mildly acidic NSAID may show some preferential distribution in inflamed tissues there was no clear experimental proof for diclofenac until the present study. PMID- 19291311 TI - ACE I/D genotype, adiposity, and blood pressure in children. AB - BACKGROUND: Angiotensin converting enzyme (ACE) is a possible candidate gene that may influence both body fatness and blood pressure. Although several genetic studies have been conducted in adults, relatively few studies have examined the contribution of potential candidate genes, and specifically ACE I/D, on adiposity and BP phenotypes in childhood. Such studies may prove insightful for the development of the obesity-hypertension phenotype early in life. The purpose of this study was to examine differences in body fatness and resting blood pressure (BP) by ACE I/D genotype, and determine if the association between adiposity and BP varies by ACE I/D genotype in children. METHODS: 152 children (75 girls, 77 boys) were assessed for body composition (% body fat) using dual energy x-ray absorbtiometry and resting BP according to American Heart Association recommendations. Buccal cell samples were genotyped using newly developed PCR RFLP tests for two SNPs (rs4341 and rs4343) in complete linkage disequilibrium with the ACE I/D polymorphism. Partial correlations were computed to assess the ociations between % body fat and BP in the total sample and by genotype. ANCOVA was used to examine differences in resting BP by ACE I/D genotype and fatness groups. RESULTS: Approximately 39% of youth were overfat based on % body fat (>30% fat in girls, 25% fat in boys). Body mass, body mass index, and fat-free mass were significantly higher in the ACE D-carriers compared to the II group (p < 0.05). BP was not significantly different by ACE I/D genotypes. In the total sample, correlations between adiposity and BP ranged from 0.30 to 0.46, and were not significantly different between genotypes. When grouped by genotype and body fat category, the overfat D-carrier subjects had significantly higher SBP and MAP compared to the normal fat D-carrier and normal fat II groups (p < 0.05). CONCLUSION: ACE D-carriers are heavier than ACE II children; however, BP did not differ by ACE I/D genotype but was adversely influenced in the overfat D carriers. Further studies are warranted to investigate the genetics of fatness and BP phenotypes in children. PMID- 19291312 TI - Divergence in gene regulation at young life history stages of whitefish (Coregonus sp.) and the emergence of genomic isolation. AB - BACKGROUND: The evolution of barriers to reproduction is of key interest to understand speciation. However, there may be a current bias towards studying intrinsic postzygotic isolation in old species pairs as compared to the emergence of barriers to gene flow through adaptive divergence. This study evaluates the relative importance of both processes in the evolution of genomic isolation in incipient species of whitefish (Coregonus clupeaformis) for which preliminary data suggest that postzygotic isolation emerges with intrinsic factors acting at embryo stages but also due to extrinsic factors during adult life. RESULTS: Gene expression data were screened using cDNA microarrays to identify regulatory changes at embryo and juvenile stages that provide evidence for genomic divergence at the underlying genetic factors. A comparison of different life history stages shows that 16-week old juvenile fish have 14 times more genes displaying significant regulatory divergence than embryos. Furthermore, regulatory changes in juvenile fish match patterns in adult fish suggesting that gene expression divergence is established early in juvenile fish and persists throughout the adult phase. Comparative analyses with results from previous studies on dwarf-normal species pairs show that at least 26 genetic factors identified in juvenile fish are candidate traits for adaptive divergence in adult fish. Eight of these show parallel directions of gene expression divergence independent of tissue type or age of the fish. The latter are associated with energy metabolism, a complex trait known to drive adaptive divergence in dwarf and normal whitefish. CONCLUSION: Although experimental evidence suggests the existence of genetic factors that cause intrinsic postzygotic isolation acting in embryos, the analysis presented here provided few candidate genes in embryos, which also corroborate previous studies showing a lack of ecological divergence between sympatric dwarf and normal whitefish at the larval stage. In contrast, gene expression divergence in juveniles can be linked to adaptive traits and seems to be driven by positive selection. The results support the idea that adaptive differentiation may be more important in explaining the emergence of barriers to gene flow in an early phase of speciation by providing a broad genomic basis for extrinsic postzygotic isolation rather than intrinsic barriers. PMID- 19291313 TI - Expression of transmembrane carbonic anhydrases, CAIX and CAXII, in human development. AB - BACKGROUND: Transmembrane CAIX and CAXII are members of the alpha carbonic anhydrase (CA) family. They play a crucial role in differentiation, proliferation, and pH regulation. Expression of CAIX and CAXII proteins in tumor tissues is primarily induced by hypoxia and this is particularly true for CAIX, which is regulated by the transcription factor, hypoxia inducible factor-1 (HIF 1). Their distributions in normal adult human tissues are restricted to highly specialized cells that are not always hypoxic. The human fetus exists in a relatively hypoxic environment. We examined expression of CAIX, CAXII and HIF 1alpha in the developing human fetus and postnatal tissues to determine whether expression of CAIX and CAXII is exclusively regulated by HIF-1. RESULTS: The co localization of CAIX and HIF-1alpha was limited to certain cell types in embryonic and early fetal tissues. Those cells comprised the primitive mesenchyma or involved chondrogenesis and skin development. Transient CAIX expression was limited to immature tissues of mesodermal origin and the skin and ependymal cells. The only tissues that persistently expressed CAIX protein were coelomic epithelium (mesothelium) and its remnants, the epithelium of the stomach and biliary tree, glands and crypt cells of duodenum and small intestine, and the cells located at those sites previously identified as harboring adult stem cells in, for example, the skin and large intestine. In many instances co-localization of CAIX and HIF-1alpha was not evident. CAXII expression is restricted to cells involved in secretion and water absorption such as parietal cells of the stomach, acinar cells of the salivary glands and pancreas, epithelium of the large intestine, and renal tubules. Co-localization of CAXII with CAIX or HIF-1alpha was not observed. CONCLUSION: The study has showed that: 1) HIF-1alpha and CAIX expression co- localized in many, but not all, of the embryonic and early fetal tissues; 2) There is no evidence of co-localization of CAIX and CAXII; 3) CAIX and CAXII expression is closely related to cell origin and secretory activity involving proton transport, respectively. The intriguing finding of rare CAIX expressing cells in those sites corresponding to stem cell niches requires further investigation. PMID- 19291314 TI - Identification of Archaea-specific chemotaxis proteins which interact with the flagellar apparatus. AB - BACKGROUND: Archaea share with bacteria the ability to bias their movement towards more favorable locations, a process known as taxis. Two molecular systems drive this process: the motility apparatus and the chemotaxis signal transduction system. The first consists of the flagellum, the flagellar motor, and its switch, which allows cells to reverse the rotation of flagella. The second targets the flagellar motor switch in order to modulate the switching frequency in response to external stimuli. While the signal transduction system is conserved throughout archaea and bacteria, the archaeal flagellar apparatus is different from the bacterial one. The proteins constituting the flagellar motor and its switch in archaea have not yet been identified, and the connection between the bacterial like chemotaxis signal transduction system and the archaeal motility apparatus is unknown. RESULTS: Using protein-protein interaction analysis, we have identified three proteins in Halobacterium salinarum that interact with the chemotaxis (Che) proteins CheY, CheD, and CheC2, as well as the flagella accessory (Fla) proteins FlaCE and FlaD. Two of the proteins belong to the protein family DUF439, the third is a HEAT_PBS family protein. In-frame deletion strains for all three proteins were generated and analyzed as follows: a) photophobic responses were measured by a computer-based cell tracking system b) flagellar rotational bias was determined by dark-field microscopy, and c) chemotactic behavior was analyzed by a swarm plate assay. Strains deleted for the HEAT_PBS protein or one of the DUF439 proteins proved unable to switch the direction of flagellar rotation. In these mutants, flagella rotate only clockwise, resulting in exclusively forward swimming cells that are unable to respond to tactic signals. Deletion of the second DUF439 protein had only minimal effects. HEAT_PBS proteins could be identified in the chemotaxis gene regions of all motile haloarchaea sequenced so far, but not in those of other archaeal species. Genes coding for DUF439 proteins, however, were found to be integral parts of chemotaxis gene regions across the archaeal domain, and they were not detected in other genomic context. CONCLUSION: Altogether, these results demonstrate that, in the archaeal domain, previously unrecognized archaea-specific Che proteins are essential for relaying taxis signaling to the flagellar apparatus. PMID- 19291315 TI - Effects of statins on the secretion of human serum albumin in cultured HepG2 cells. AB - Statins reduce cholesterol biosynthesis by inhibiting HMG-CoA reductase and thereby lower total cholesterol and LDL cholesterol levels in serum, which in turn lower the incidence of cardiovascular disease (CVD). Statins are also known to modulate various cellular functions such as gene expression, cell proliferation, and programmed cell death through inhibition of downstream intermediates in cholesterol synthesis. In this study, we have investigated the possible effects of statins on the secretion of serum albumin from cultured HepG2 cells since high levels of serum albumin are associated with reduced risks for CVD and statins are effective in lowering the risk of CVD through other effects in addition to their effects on serum total cholesterol and LDL cholesterol levels, known as pleiotropic effects. Our results showed that simvastatin increased HSA secretion up to 32.3% compared to the control group. Among 3 statin analogs we tested, simvastatin exhibited the highest stimulatory effects on HSA secretion compared to the control group. Our study also showed that the increased HSA secretions from HepG2 cells by simvastatin treatments were due to the increased rate of HSA synthesis, not due to the reduced posttranslational degradation rate of HSA. Our finding suggests another added benefit of statins' treatments in preventing CVD through stimulation of HSA biosynthesis. PMID- 19291316 TI - Transcriptomic basis for an antiserum against Micrurus corallinus (coral snake) venom. AB - BACKGROUND: Micrurus corallinus (coral snake) is a tropical forest snake belonging to the family Elapidae. Its venom shows a high neurotoxicity associated with pre- and post-synaptic toxins, causing diaphragm paralysis, which may result in death. In spite of a relatively small incidence of accidents, serum therapy is crucial for those bitten. However, the adequate production of antiserum is hampered by the difficulty in obtaining sufficient amounts of venom from a small snake with demanding breeding conditions. In order to elucidate the molecular basis of this venom and to uncover possible immunogens for an antiserum, we generated expressed sequences tags (ESTs) from its venom glands and analyzed the transcriptomic profile. In addition, their immunogenicity was tested using DNA immunization. RESULTS: A total of 1438 ESTs were generated and grouped into 611 clusters. Toxin transcripts represented 46% of the total ESTs. The two main toxin classes consisted of three-finger toxins (3FTx) (24%) and phospholipases A(2) (PLA(2)s) (15%). However, 8 other classes of toxins were present, including C type lectins, natriuretic peptide precursors and even high-molecular mass components such as metalloproteases and L-amino acid oxidases. Each class included an assortment of isoforms, some showing evidence of alternative splicing and domain deletions. Five antigenic candidates were selected (four 3FTx and one PLA(2)) and used for a preliminary study of DNA immunization. The immunological response showed that the sera from the immunized animals were able to recognize the recombinant antigens. CONCLUSION: Besides an improvement in our knowledge of the composition of coral snake venoms, which are very poorly known when compared to Old World elapids, the expression profile suggests abundant and diversified components that may be used in future antiserum formulation. As recombinant production of venom antigens frequently fails due to complex disulfide arrangements, DNA immunization may be a viable alternative. In fact, the selected candidates provided an initial evidence of the feasibility of this approach, which is less costly and not dependent on the availability of the venom. PMID- 19291317 TI - Protective effects of total fraction of avocado/soybean unsaponifiables on the structural changes in experimental dog osteoarthritis: inhibition of nitric oxide synthase and matrix metalloproteinase-13. AB - INTRODUCTION: The aims of this study were, first, to investigate the in vivo effects of treatment with avocado/soybean unsaponifiables on the development of osteoarthritic structural changes in the anterior cruciate ligament dog model and, second, to explore their mode of action. METHODS: Osteoarthritis was induced by anterior cruciate ligament transection of the right knee in crossbred dogs. There were two treatment groups (n = 8 dogs/group), in which the animals received either placebo or avocado/soybean unsaponifiables (10 mg/kg per day), which were given orally for the entire duration of the study (8 weeks). We conducted macroscopic and histomorphological analyses of cartilage and subchondral bone of the femoral condyles and/or tibial plateaus. We also conducted immunohistochemical analyses in cartilage for the following antigens: inducible nitric oxide synthase, matrix metalloproteinase (MMP)-1, MMP-13, a disintegrin and metalloproteinase domain with thrombospondin motifs (ADAMTS)4 and ADAMTS5. RESULTS: The size of macroscopic lesions on the tibial plateaus was decreased (P = 0.04) in dogs treated with the avocado/soybean unsaponifiables. Histologically, in these animals the severity of cartilage lesions on both tibial plateaus and femoral condyles, and the cellular infiltration in synovium were significantly decreased (P = 0.0002 and P = 0.04, respectively). Treatment with avocado/soybean unsaponifiables also reduced loss of subchondral bone volume (P < 0.05) and calcified cartilage thickness (P = 0.01) compared with placebo. Immunohistochemical analysis of cartilage revealed that avocado/soybean unsaponifiables significantly reduced the level of inducible nitric oxide synthase (P < 0.05) and MMP-13 (P = 0.01) in cartilage. CONCLUSIONS: This study demonstrates that treatment with avocado/soybean unsaponifiables can reduce the development of early osteoarthritic cartilage and subchondral bone lesions in the anterior cruciate ligament dog model of osteoarthritis. This effect appears to be mediated through the inhibition of inducible nitric oxide synthase and MMP-13, which are key mediators of the structural changes that take place in osteoarthritis. PMID- 19291318 TI - Increasingly transformed MCF-10A cells have a progressively tumor-like phenotype in three-dimensional basement membrane culture. AB - BACKGROUND: MCF-10A cells are near diploid and normal human mammary epithelial cells. In three-dimensional reconstituted basement membrane culture, they undergo a well-defined program of proliferation, differentiation, and growth arrest, forming acinar structures that recapitulate many aspects of mammary architecture in vivo. The pre-malignant MCF-10AT cells and malignant MCF-10CA1a lines were sequentially derived from the MCF-10A parental cell line first by expression of a constitutively active T24 H-Ras generating the MCF-10AT cell line. This was followed by repeated selection for increasingly aggressive tumor formation from cells recovered from xenograft tumors in immuno-compromised mice, generating the MCF-10CA1a cell line. When inoculated subcutaneously into the flanks of immuno compromised mice, MCF-10AT cells occasionally form tumors, whereas MCF-10CA1a cells invariably form tumors with a shorter latency than MCF-10AT derived tumors. RESULTS: MCF-10AT cells grown in three-dimensional basement membrane culture form complex multi-acinar structures that produce a basement membrane but undergo delayed cell cycle arrest and have incomplete luminal development. MCF-10CA1a cells grown in three-dimensional basement membrane culture form large, hyper proliferative masses, that retain few characteristics of MCF10A acini and more closely resemble tumors. CONCLUSION: Here we report on the growth and differentiation properties of these three matched cell lines in three-dimensional basement membrane culture. Features of tissue morphogenesis were assessed, including proliferation, basement membrane formation, polarization of alpha-6 beta-4 integrin to the basement membrane, formation of cell:cell junctions, and apoptosis for luminal clearance. The matched series of normal MCF-10A, pre malignant MCF-10AT, and malignant MCF-10CA1a cells offers a unique opportunity to study the mechanisms of malignant progression both in a three-dimensional microenvironment and in the same cell background. PMID- 19291319 TI - The PI3K-Akt-mTOR pathway regulates Abeta oligomer induced neuronal cell cycle events. AB - Accumulating evidence suggests that neurons prone to degeneration in Alzheimer's Disease (AD) exhibit evidence of re-entry into an aberrant mitotic cell cycle. Our laboratory recently demonstrated that, in a genomic amyloid precursor protein (APP) mouse model of AD (R1.40), neuronal cell cycle events (CCEs) occur in the absence of beta-amyloid (Abeta) deposition and are still dependent upon the amyloidogenic processing of the amyloid precursor protein (APP). These data suggested that soluble Abeta species might play a direct role in the induction of neuronal CCEs. Here, we show that exposure of non-transgenic primary cortical neurons to Abeta oligomers, but not monomers or fibrils, results in the retraction of neuronal processes, and induction of CCEs in a concentration dependent manner. Retraction of neuronal processes correlated with the induction of CCEs and the Abeta monomer or Abeta fibrils showed only minimal effects. In addition, we provide evidence that induction of neuronal CCEs are autonomous to primary neurons cultured from the R1.40 mice. Finally, our results also demonstrate that Abeta oligomer treated neurons exhibit elevated levels of activated Akt and mTOR (mammalian Target Of Rapamycin) and that PI3K, Akt or mTOR inhibitors blocked Abeta oligomer-induced neuronal CCEs. Taken together, these results demonstrate that Abeta oligomer-based induction of neuronal CCEs involve the PI3K-Akt-mTOR pathway. PMID- 19291320 TI - Expression-based discovery of candidate ovule development regulators through transcriptional profiling of ovule mutants. AB - BACKGROUND: Arabidopsis ovules comprise four morphologically distinct parts: the nucellus, which contains the embryo sac, two integuments that become the seed coat, and the funiculus that anchors the ovule within the carpel. Analysis of developmental mutants has shown that ovule morphogenesis relies on tightly regulated genetic interactions that can serve as a model for developmental regulation. Redundancy, pleiotropic effects and subtle phenotypes may preclude identification of mutants affecting some processes in screens for phenotypic changes. Expression-based gene discovery can be used access such obscured genes. RESULTS: Affymetrix microarrays were used for expression-based gene discovery to identify sets of genes expressed in either or both integuments. The genes were identified by comparison of pistil mRNA from wild type with mRNA from two mutants; inner no outer (ino, which lacks the outer integument), and aintegumenta (ant, which lacks both integuments). Pools of pistils representing early and late stages of ovule development were evaluated and data from the three genotypes were used to designate genes that were predominantly expressed in the integuments using pair-wise and cluster analyses. Approximately two hundred genes were found to have a high probability of preferential expression in these structures, and the predictive nature of the expression classes was confirmed with reverse transcriptase polymerase chain reaction and in situ hybridization. CONCLUSION: The results showed that it was possible to use a mutant, ant, with broad effects on plant phenotype to identify genes expressed specifically in ovules, when coupled with predictions from known gene expression patterns, or in combination with a more specific mutant, ino. Robust microarray averaging (RMA) analysis of array data provided the most reliable comparisons, especially for weakly expressed genes. The studies yielded an over-abundance of transcriptional regulators in the identified genes, and these form a set of candidate genes for evaluation of roles in ovule development using reverse genetics. PMID- 19291321 TI - Elevated antiphospholipid antibody titers and adverse pregnancy outcomes: analysis of a population-based hospital dataset. AB - BACKGROUND: The primary objective of this study was to determine if elevated antiphospholipid antibody titers were correlated with the presence of preeclampsia/eclampsia, systemic lupus erythematosus (SLE), placental insufficiency, and a prolonged length of stay (PLOS), in women who delivered throughout Florida, USA. METHODS: Cross-sectional analyses were conducted using a statewide hospital database. Prevalence odds ratios (OR) were calculated to quantify the association between elevated antiphospholipid antibody titers and four outcomes in 141,286 women who delivered in Florida in 2001. The possibility that the relationship between elevated antiphospholipid antibody titers and the outcomes of preeclampsia/eclampsia, placental insufficiency, and PLOS, may have been modified by the presence of SLE was evaluated in a multiple logistic regression model by creating a composite interaction term. RESULTS: Women with elevated antiphospholipid antibody titers (n = 88) were older, more likely to be of white race and not on Medicaid than women who did not have elevated antiphospholipid antibody titers. Women who had elevated antiphospholipid antibody titers had an increased adjusted odds ratio for preeclampsia and eclampsia, (OR = 2.93 p = 0.0015), SLE (OR = 61.24 p < 0.0001), placental insufficiency (OR = 4.58 p = 0.0003), and PLOS (OR = 3.93 p < 0.0001). Patients who had both an elevated antiphospholipid antibody titer and SLE were significantly more likely than the comparison group (women without an elevated titer who did not have SLE) to have the outcomes of preeclampsia, placental insufficiency and PLOS. CONCLUSION: This exploratory epidemiologic investigation found moderate to very strong associations between elevated antiphospholipid antibody titers and four important outcomes in a large sample of women. PMID- 19291322 TI - Acetaminophen inhibits neuronal inflammation and protects neurons from oxidative stress. AB - BACKGROUND: Recent studies have demonstrated a link between the inflammatory response, increased cytokine formation, and neurodegeneration in the brain. The beneficial effects of anti-inflammatory drugs in neurodegenerative diseases, such as Alzheimer's disease (AD), have been documented. Increasing evidence suggests that acetaminophen has unappreciated anti-oxidant and anti-inflammatory properties. The objectives of this study are to determine the effects of acetaminophen on cultured brain neuronal survival and inflammatory factor expression when exposed to oxidative stress. METHODS: Cerebral cortical cultured neurons are pretreated with acetaminophen and then exposed to the superoxide generating compound menadione (5 microM). Cell survival is assessed by MTT assay and inflammatory protein (tumor necrosis factor alpha, interleukin-1, macrophage inflammatory protein alpha, and RANTES) release quantitated by ELISA. Expression of pro- and anti-apoptotic proteins is assessed by western blots. RESULTS: Acetaminophen has pro-survival effects on neurons in culture. Menadione, a superoxide releasing oxidant stressor, causes a significant (p < 0.001) increase in neuronal cell death as well as in the release of tumor necrosis factor alpha, interleukin-1, macrophage inflammatory protein alpha, and RANTES from cultured neurons. Pretreatment of neuronal cultures with acetaminophen (50 microM) increases neuronal cell survival and inhibits the expression of these cytokines and chemokines. In addition, we document, for the first time, that acetaminophen increases expression of the anti-apoptotic protein Bcl2 in brain neurons and decreases the menadione-induced elevation of the proapoptotic protein, cleaved caspase 3. We show that blocking acetaminophen-induced expression of Bcl2 reduces the pro-survival effect of the drug. CONCLUSION: These data show that acetaminophen has anti-oxidant and anti-inflammatory effects on neurons and suggest a heretofore unappreciated therapeutic potential for this drug in neurodegenerative diseases such as AD that are characterized by oxidant and inflammatory stress. PMID- 19291323 TI - Fever, hyperglycaemia and swallowing dysfunction management in acute stroke: a cluster randomised controlled trial of knowledge transfer. AB - BACKGROUND: Hyperglycaemia, fever, and swallowing dysfunction are poorly managed in the admission phase of acute stroke, and patient outcomes are compromised. Use of evidence-based guidelines could improve care but have not been effectively implemented. Our study aims to develop and trial an intervention based on multidisciplinary team-building to improve management of fever, hyperglycaemia, and swallowing dysfunction in patients following acute stroke. METHODS AND DESIGN: Metropolitan acute stroke units (ASUs) located in New South Wales, Australia will be stratified by service category (A or B) and, within strata, by baseline patient recruitment numbers (high or low) in this prospective, multicentre, single-blind, cluster randomised controlled trial (CRCT). ASUs then will be randomised independently to either intervention or control groups. ASUs allocated to the intervention group will receive: unit-based workshops to identify local barriers and enablers; a standardised core education program; evidence-based clinical treatment protocols; and ongoing engagement of local staff. Control group ASUs will receive only an abridged version of the National Clinical Guidelines for Acute Stroke Management. The following outcome measures will be collected at 90 days post-hospital admission: patient death, disability (modified Rankin Score); dependency (Barthel Index) and Health Status (SF-36). Additional measures include: performance of swallowing screening within 24 hours of admission; glycaemic control and temperature control. DISCUSSION: This is a unique study of research transfer in acute stroke. Providing optimal inpatient care during the admission phase is essential if we are to combat the rising incidence of debilitating stroke. Our CRCT will also allow us to test interventions focussed on multidisciplinary ASU teams rather than individual disciplines, an imperative of modern hospital services. TRIAL REGISTRATION: Australia New Zealand Clinical Trial Registry (ANZCTR) No: ACTRN12608000563369. PMID- 19291324 TI - Time spent playing outdoors after school and its relationship with independent mobility: a cross-sectional survey of children aged 10-12 years in Sydney, Australia. AB - BACKGROUND: Time spent outdoors is positively associated with physical activity and has been suggested as a proxy for physical activity of children. The role of children's independence in physical activity and time spent outdoors is less understood. This study aimed to assess how much time children spent playing outdoors after school, and to explore the relationship between outdoor play and independence among children aged 10-12 years. METHOD: Children recorded how much time they spent playing outdoors or watching TV/videos or playing computer games after school using a five-day diary, and also reported whether they were allowed to walk on their own in their neighbourhood as an indicator of their independent mobility. Parents were surveyed on family demographics and perception of neighbourhood safety. The surveys were conducted in late 2006 as part of the Central Sydney Walk to School program which involved 1975 children and their parents from 24 primary schools. Factors associated with time spent playing outdoors were determined by logistic regression modelling. RESULTS: Thirty-seven per cent of children spent less than half an hour a day playing outdoors after school, and 43% spent more than 2 hours a day watching TV, videos or playing computer games. Forty-eight per cent of children were allowed to walk on their own near where they lived. Children's independent mobility was significantly associated with outdoor play after adjusting for other confounders. Compared with those who were never allowed to walk on their own near where they lived, students who were allowed to walk on their own were significantly more likely to spend more than half an hour a day playing outdoors after school with an adjusted odds ratio of 2.6, 95% CI 1.84-3.58, P < 0.001. CONCLUSION: The findings that a significant proportion of children spend less than half an hour a day playing outdoors after school and have excessive screen time have important implications for physical activity promotion and obesity prevention. The study also suggests that children's independent mobility should be considered in research and evaluation into children's play and physical activity. Environments that promote greater independent mobility in children may increase their physical activity levels and hence reduce their risk of overweight/obesity. PMID- 19291325 TI - Procalcitonin kinetics within the first days of sepsis: relationship with the appropriateness of antibiotic therapy and the outcome. AB - INTRODUCTION: Management of the early stage of sepsis is a critical issue. As part of it, infection control including appropriate antibiotic therapy administration should be prompt. However, microbiological findings, if any, are generally obtained late during the course of the disease. The potential interest of procalcitonin (PCT) as a way to assess the clinical efficacy of the empirical antibiotic therapy was addressed in the present study. METHODS: An observational cohort study including 180 patients with documented sepsis was conducted in our 15-bed medical intensive care unit (ICU). Procalcitonin measurement was obtained daily over a 4-day period following the onset of sepsis (day 1 (D1) to D4). The PCT time course was analyzed according to the appropriateness of the first-line empirical antibiotic therapy as well as according to the patient outcome. RESULTS: Appropriate first-line empirical antibiotic therapy (n = 135) was associated with a significantly greater decrease in PCT between D2 and D3 (DeltaPCT D2-D3) (-3.9 (35.9) vs. +5.0 (29.7), respectively; P < 0.01). In addition, DeltaPCT D2-D3 was found to be an independent predictor of first-line empirical antibiotic therapy appropriateness. In addition, a trend toward a greater rise in PCT between D1 and D2 was observed in patients with inappropriate antibiotics as compared with those with appropriate therapy (+5.2 (47.4) and +1.7 (35.0), respectively; P = 0.20). The D1 PCT level failed to predict outcome, but higher levels were measured in the nonsurvivors (n = 51) when compared with the survivors (n = 121) as early as D3 (40.8 (85.7) and 21.3 (41.0), respectively; P = 0.04). Moreover, PCT kinetics between D2 and D3 were also found to be significantly different, since a decrease >or= 30% was expected in the survivors (log-rank test, P = 0.04), and was found to be an independent predictor of survival (odds ratio = 2.94; 95% confidence interval 1.22 to 7.09; P = 0.02). CONCLUSIONS: In our study in an ICU, appropriateness of the empirical antibiotic therapy and the overall survival were associated with a greater decline in PCT between D2 and D3. Further studies are needed to assess the utility of the daily monitoring of PCT in addition to clinical evaluation during the early management of sepsis. PMID- 19291326 TI - Spatial analysis of falls in an urban community of Hong Kong. AB - BACKGROUND: Falls are an issue of great public health concern. This study focuses on outdoor falls within an urban community in Hong Kong. Urban environmental hazards are often place-specific and dependent upon the built features, landscape characteristics, and habitual activities. Therefore, falls must be examined with respect to local situations. RESULTS: This paper uses spatial analysis methods to map fall occurrences and examine possible environmental attributes of falls in an urban community of Hong Kong. The Nearest neighbour hierarchical (Nnh) and Standard Deviational Ellipse (SDE) techniques can offer additional insights about the circumstances and environmental factors that contribute to falls. The results affirm the multi-factorial nature of falls at specific locations and for selected groups of the population. CONCLUSION: The techniques to detect hot spots of falls yield meaningful results that enable the identification of high risk locations. The combined use of descriptive and spatial analyses can be beneficial to policy makers because different preventive measures can be devised based on the types of environmental risk factors identified. The analyses are also important preludes to establishing research hypotheses for more focused studies. PMID- 19291327 TI - Training health care workers to promote HIV services for patients with tuberculosis in the Democratic Republic of Congo. AB - BACKGROUND: HIV counseling and testing, HIV prevention and provision of HIV care and support are essential activities to reduce the burden of HIV among patients with TB, and should be integrated into routine TB care. METHODS: The development of training materials to promote HIV services for TB patients involved the definition of target health care workers (HCWs); identification of required tasks, skills and knowledge; review of international guidelines; and adaptation of existing training materials for voluntary counseling and testing, prevention of mother-to-child transmission of HIV, and management of opportunistic infections (OIs). Training effectiveness was assessed by means of questionnaires administered pre- and post-training, by correlating post-training results of HCWs with the centre's HIV testing acceptance rates, and through participatory observations at the time of on-site supervisory visits and monthly meetings. RESULTS: Pre-training assessment identified gaps in basic knowledge of HIV epidemiology, the link between TB and HIV, interpretation of CD4 counts, prevention and management of OIs, and occupational post-exposure prophylaxis (PEP). Opinions on patients' rights and confidentiality varied. Mean test results increased from 72% pre-training to 87% post-training (p<0.001). Important issues regarding HIV epidemiology and PEP remained poorly understood post-training. Mean post-training scores of clinic's HCWs were significantly correlated with the centre's HIV testing acceptance rates (p=0.01). On-site supervisory visits and monthly meetings promoted staff motivation, participatory problem solving and continuing education. Training was also used as an opportunity to improve patient centred care and HCWs' communication skills. CONCLUSION: Many HCWs did not possess the knowledge or skills necessary to integrate HIV activities into routine care for patients with TB. A participatory approach resulted in training materials that fulfilled local needs. PMID- 19291328 TI - Duck (Anas platyrhynchos) linkage mapping by AFLP fingerprinting. AB - Amplified fragment length polymorphism (AFLP) with multicolored fluorescent molecular markers was used to analyze duck (Anas platyrhynchos) genomic DNA and to construct the first AFLP genetic linkage map. These markers were developed and genotyped in 766 F2 individuals from six families from a cross between two different selected duck lines, brown Tsaiya and Pekin. Two hundred and ninety-six polymorphic bands (64% of all bands) were detected using 18 pairs of fluorescent TaqI/EcoRI primer combinations. Each primer set produced a range of 7 to 29 fragments in the reactions, and generated on average 16.4 polymorphic bands. The AFLP linkage map included 260 co-dominant markers distributed in 32 linkage groups. Twenty-one co-dominant markers were not linked with any other marker. Each linkage group contained three to 63 molecular markers and their size ranged between 19.0 cM and 171.9 cM. This AFLP linkage map provides important information for establishing a duck chromosome map, for mapping quantitative trait loci (QTL mapping) and for breeding applications. PMID- 19291330 TI - Ultrasound-assisted percutaneous trigger finger release: it is safe. PMID- 19291329 TI - From bench to bedside: current and future applications of molecular profiling in renal cell carcinoma. AB - Among the adult population, renal cell carcinoma (RCC) constitutes the most prevalent form of kidney neoplasm. Unfortunately, RCC is relatively asymptomatic and there are no tumor markers available for diagnostic, prognostic or predictive purposes. Molecular profiling, the global analysis of gene and protein expression profiles, is an emerging promising tool for new biomarker identification in RCC. In this review, we summarize the existing knowledge on RCC regarding clinical presentation, treatment options, and tumor marker status. We present a general overview of the more commonly used approaches for molecular profiling at the genomic, transcriptomic and proteomic levels. We also highlight the emerging role of molecular profiling as not only revolutionizing the process of new tumor marker discovery, but also for providing a better understanding of the pathogenesis of RCC that will pave the way towards new targeted therapy discovery. Furthermore, we discuss the spectrum of clinical applications of molecular profiling in RCC in the current literature. Finally, we highlight some of the potential challenging that faces the era of molecular profiling and its transition into clinical practice, and provide an insight about the future perspectives of molecular profiling in RCC. PMID- 19291331 TI - The tag test for Dupuytren's surgery. AB - Digital nerve injury is a well-recognised complication of surgical treatment for Dupuytren's disease. We describe a simple test, the Tag test, that can be used intra-operatively to help identification of the digital nerves. PMID- 19291332 TI - Medication discrepancies upon hospital to skilled nursing facility transitions. AB - BACKGROUND: Failure to reconcile medications across transitions in care is an important source of harm to patients. Little is known about medication discrepancies upon admission to skilled nursing facilities (SNFs). OBJECTIVE: To describe the prevalence of, type of medications involved in, and sources of medication discrepancies upon admission to the SNF setting. DESIGN: Cross sectional study. PARTICIPANTS: Patients admitted to SNF for subacute care. MEASUREMENTS: Number of medication discrepancies, defined as unexplained differences among documented medication regimens, including the hospital discharge summary, patient care referral form and SNF admission orders. RESULTS: Of 2,319 medications reviewed on admission, 495 (21.3%) had a medication discrepancy. At least one medication discrepancy was identified in 142 of 199 (71.4%) SNF admissions. The discharge summary and the patient care referral form did not match in 104 of 199 (52.3%) SNF admissions. Disagreement between the discharge summary and the patient care referral form accounted for 62.0% (n = 307) of all medication discrepancies. Cardiovascular agents, opioid analgesics, neuropsychiatric agents, hypoglycemics, antibiotics, and anticoagulants accounted for over 50% of all discrepant medications. CONCLUSIONS: Medication discrepancies occurred in almost three out of four SNF admissions and accounted for one in five medications prescribed on admission. The discharge summary and the patient care referral forms from the discharging institution are often in disagreement. Our study findings underscore the importance of current efforts to improve the quality of inter-institutional communication. PMID- 19291334 TI - Accurate preoperative localization of insulinomas avoids the need for blind resection and reoperation: analysis of a single institution experience with 17 surgically treated tumors over 19 years. AB - INTRODUCTION: Presently, the need for and choice of preoperative localization tests for insulinomas remain controversial. We report the results from a single institution experience whereby the management policy adopted was that of accurate preoperative localization before surgical exploration. MATERIALS AND METHODS: From 1990 to 2008, 17 patients with a clinical and biochemical diagnosis of an insulinoma who underwent surgery were retrospectively reviewed. The diagnosis of all insulinomas were confirmed pathologically. RESULTS: All tumors were localized preoperatively and an average of 2.2 preoperative localization studies including 1.4 noninvasive studies and 0.8 invasive studies were utilized per patient. Invasive localization modalities were more sensitive (92%) than noninvasive modalities in localizing insulinomas (71%). Intra-arterial calcium stimulation with hepatic venous sampling was the most sensitive invasive modality (100%), whereas magnetic resonance imaging was the most sensitive noninvasive modality (63%). Fifteen of 17 tumors (88%) were localized intraoperatively via inspection/palpation and/or intraoperative ultrasonography. Both insulinomas which were not localized intraoperatively were localized correctly to the distal pancreas via preoperative transhepatic portal venous sampling. None of the patients required a blind resection or surgical reexploration for failed localization. All 17 patients underwent complete surgical resection which included eight enucleations and nine distal pancreatectomies with a cure rate of 94% (16/17) at a median follow-up of 35 (range, 1-217) months. The postoperative morbidity and long-term outcome of enucleation was similar to distal pancreatectomy despite a higher rate of microscopic margin involvement. CONCLUSION: Accurate preoperative localization of insulinomas is useful as it eliminates the need for blind distal pancreatectomy and avoids reoperation. Complete surgical resection is the treatment of choice, and whenever possible, a pancreas-sparing approach such as enucleation should be adopted. PMID- 19291335 TI - The omentum: anatomical, metabolic, and surgical aspects. AB - INTRODUCTION: The omentum is acknowledged to have diverse functions in the pathophysiology of intra-abdominal disease. Its angiogenic properties act as a natural defense mechanism in peritonitis and intra-abdominal sepsis. With advancing technology the omentum is revealing itself as a new player in the field of molecular surgery with special reference to cancer, obesity and tissue reconstruction. MATERIALS AND METHODS: This article reviews the existing and potential surgical applications of the omentum. PMID- 19291336 TI - Prevalence of internal hernias after laparoscopic colonic surgery. AB - INTRODUCTION: Laparoscopic approach for colorectal resections is gaining popularity. Internal small bowel herniation (SBH) through a mesenteric defect has been described and, although rare, is a severe complication. The aim of this study was to evaluate the incidence and outcome of internal hernias after laparoscopic colorectal resection. MATERIAL AND METHODS: During a 5-year period, all patients who underwent laparoscopic left colon resection were included in the study. A retrospective data base query was performed searching for all patients in whom SBH required surgical reintervention. RESULTS: A total of 436 laparoscopic left colorectal resections were performed from January 2000 to July 2006. Five male patients presented symptomatic internal hernias and required re operation. Four had a resection for cancer and one for sigmoiditis. The mesenteric defect was not initially closed in three cases. In all cases, we found small bowel hernias through the mesocolon defect. One patient was re-operated on post-op day 2 for mesenteric ischemia and died after 24 h. DISCUSSION: Internal hernia is a rare but fatal complication after laparoscopic colonic resection. Suspicion of this diagnosis requires emergency re-operation because symptoms are nonspecific. CONCLUSION: All mesenteric defects created during colorectal laparoscopy surgery should be meticulously closed. PMID- 19291337 TI - Impact of KIT and PDGFRA gene mutations on prognosis of patients with gastrointestinal stromal tumors after complete primary tumor resection. AB - INTRODUCTION: To investigate the impact of KIT and PDGFRA gene mutations on the prognosis of gastrointestinal stromal tumors (GIST). MATERIAL AND METHODS: Tumor tissue from 184 patients with primary GIST was submitted to mutational analysis of exons 9, 11, 13, and 17 of the KIT gene and exons 12 and 18 of the PDGFRA gene. Clinical and pathological parameters were analyzed and correlated to the risk of recurrence and disease-free survival (DFS). RESULTS AND DISCUSSION: The authors found that somatic mutations were detected in 162 tumors (88.0%). Age, clinical stage, mitotic count, and tumor size were of prognostic relevance on both univariate and multivariate analysis. Five-year DFS was 41.9%. While the presence of a KIT or PDGFRA mutation per se was not associated with tumor recurrence and/or disease-free survival, exon 11 deletion and hemizygous mutation status were both independent factors highly predictive for poor survival. CONCLUSION: The authors conclude that KIT exon 11 deletions and somatic loss of the wild-type KIT identified patients with poor prognosis. Age, clinical stage, tumor size, and mitotic count were standard clinicopathologic features that significantly influenced the prognosis. Mutation type of the mitogen receptor c kit has a potential for predicting the course of the disease and might contribute to management individualization of GIST patients. PMID- 19291338 TI - Analysis of safety and efficacy of intragastric balloon in extremely obese patients. AB - BACKGROUND: The implantation of an intragastric balloon constitutes a short-term effective non-surgical intervention to lose weight. The aim of this study was to evaluate retrospectively the clinical outcome and safety of gastric balloon therapy (GBT) in extremely obese patients. METHODS: One hundred and nine super- and super-super-obese patients, 64 males and 45 females, mean age 39.1+/-8.4 years, mean body mass index (BMI) 68.8+/-8.9 kg/m2, who underwent GBT for weight loss, were studied retrospectively. GBT was assessed in massively obese patients concerning tolerance, weight loss, number of comorbidities and complications. RESULTS: A significant reduction in patients' weight and BMI was evident after GBT. Regarding safety, no major complications occurred. Minor complications at balloon placement and removal occurred in one (0.9%) and three patients (2.8%) respectively. Mean duration of GBT was 177.6+/-56.8 days. After GBT, the mean weight loss was 26.3+/-15.2 kg (p<0.001) and the mean BMI reduction was 8.7+/-5.1 kg/m2 (p<0.001) representing a mean percentage of excess BMI lost (%EBL) of 19.7+/-10.2. The highest BMI loss was observed in patients with BMI>80 kg/m2. A noteworthy improvement of comorbidities in 56.8% of the patients was also noted. Of the 109 patients, 69 received subsequent bariatric surgery. All the procedures were performed laparoscopically. Ten patients, with a mean BMI of 68.6+/-10.6 kg/m2 after the removal of the first BIB, received a second BIB resulting in a non-significant weight and BMI loss of 6.3+/-9.4 kg and 1.8+/-2.9 kg/m2, respectively. CONCLUSIONS: Our study indicates the safety and efficacy of GBT in extremely obese patients particularly as a first step before a definitive anti obesity operation. GBT appears to be a safe, tolerable, and potentially effective procedure for the initial treatment of morbid obesity. PMID- 19291339 TI - After 3 years of starvation: duodenum swallowed remaining stomach. AB - A 42-year-old morbidly obese patient (BMI 44.1 kg/m(2)) was admitted to our emergency room with upper abdominal pain, nausea, and cholestasis. Nine years ago, a vertical banded gastroplasty had been performed (former BMI 53.5 kg/m(2)) with a subsequent weight loss to BMI 33.0 kg/m(2). After regaining weight up to a BMI of 47.6 kg/m(2), 5 years ago a conversion to a gastric bypass was realized. A computed tomography of the abdomen showed an invagination of the remaining stomach into the duodenum causing obstruction of the orifice of common bile duct. The patient underwent an open desinvagination of the intussusception and resection of the remaining stomach. Gastroduodenal intussusception is rare and mostly secondary to gastric lipoma. To prevent this rare but serious complication, the remaining stomach could be fixed at the crura of the diaphragm, tagged to the anterior abdominal wall by temporary gastrostomy tube, or resected. PMID- 19291340 TI - What carries a mediation process? Configural analysis of mediation. AB - Mediation is a process that links a predictor and a criterion via a mediator variable. Mediation can be full or partial. This well-established definition operates at the level of variables even if they are categorical. In this article, two new approaches to the analysis of mediation are proposed. Both of these approaches focus on the analysis of categorical variables. The first involves mediation analysis at the level of configurations instead of variables. Thus, mediation can be incorporated into the arsenal of methods of analysis for person oriented research. Second, it is proposed that Configural Frequency Analysis (CFA) can be used for both exploration and confirmation of mediation relationships among categorical variables. The implications of using CFA are first that mediation hypotheses can be tested at the level of individual configurations instead of variables. Second, this approach leaves the door open for different types of mediation processes to exist within the same set. Using a data example, it is illustrated that aggregate-level analysis can overlook mediation processes that operate at the level of individual configurations. PMID- 19291342 TI - Costing behavioral interventions: a practical guide to enhance translation. AB - BACKGROUND/PURPOSE: Cost and cost effectiveness of behavioral interventions are critical parts of dissemination and implementation into non-academic settings. Due to the lack of indicative data and policy makers' increasing demands for both program effectiveness and efficiency, cost analyses can serve as valuable tools in the evaluation process. METHODS: To stimulate and promote broader use of practical techniques that can be used to efficiently estimate the implementation costs of behavioral interventions, we propose a set of analytic steps that can be employed across a broad range of interventions. RESULTS/CONCLUSIONS: Intervention costs must be distinguished from research, development, and recruitment costs. The inclusion of sensitivity analyses is recommended to understand the implications of implementation of the intervention into different settings using different intervention resources. To illustrate these procedures, we use data from a smoking reduction practical clinical trial to describe the techniques and methods used to estimate and evaluate the costs associated with the intervention. Estimated intervention costs per participant were $419, with a range of $276 to $703, depending on the number of participants. PMID- 19291343 TI - Bone microdamage: a clinical perspective. AB - INTRODUCTION: Microdamage accumulation due to fatigue loading may lead to fracture. In addition, several studies using animal models have suggested in recent years that bisphosphonates might increase microdamage accumulation. METHODS: We have reviewed the literature after a PubMed search, to examine the techniques to look for microcracks, the relationship between microdamage and bone strength, and the influence of anti-osteoporosis agents. RESULTS: Currently, the search for microcracks relies on bulk staining of bone samples, which are then examined on optic microscopy and fluorescence or confocal microscopy. The accumulation of microdamage is associated with fatigue loading and is likely to trigger targeted bone remodeling, especially in cortical bone. Several studies examining beagle dogs receiving bisphosphonates have shown a dose-dependent accumulation of microdamage in bone, with conflicting results regarding the consequences on bone mechanical properties. In living humans, obtaining data is limited to the iliac crest bone. The potential association between long-term bisphosphonate use and microcrack accumulation at the iliac crest bone has not been established unequivocally. CONCLUSIONS: Bone microdamage is critical in the understanding of bone quality. Assessment of microdamage is technically difficult, especially in humans. The clinical impact of microdamage potentially induced by bone drugs has not been established in humans. PMID- 19291345 TI - Translating research into clinical practice. Abstracts of the Eighth International Symposium on Osteoporosis. April 1-5, 2009. Washington, DC, USA. PMID- 19291344 TI - BMD, clinical risk factors and their combination for hip fracture prevention. AB - SUMMARY: This study examined the effects of the use of clinical risk factors (CRFs) alone, BMD alone or the combination using the FRAX tool for the detection of women at risk of hip fracture. BMD tests alone selected women at higher risk and a greater number of hip fracture cases were identified compared to the use of CRFs alone. The combined use of CRFs and BMD identified fewer women above a threshold risk than the use of BMD alone, but with a higher hip fracture risk and thus had the more favourable positive predictive value (PPV) and number needed to treat (NNT). INTRODUCTION: Algorithms have recently become available for the calculation of hip fracture probability from CRFs with and without information on femoral neck BMD. The aim of this study was to examine the effects of the use of CRFs alone, BMD alone or their combination using the FRAX tool for the detection of women at risk of hip fracture. METHODS: Data from 10 prospective population based cohorts, in which BMD and CRFs were documented, were used to compute the 10 year probabilities of hip fracture calibrated to the fracture and death hazards of the UK. The effects of the use of BMD tests were examined in simulations where BMD tests were used alone, CRFs alone or their combined use. The base case examined the effects in women at the age of 65 years. The principal outcome measures were the number of women identified above an intervention threshold, the number of hip fracture cases that would be identified, the positive predicted value and the NNT to prevent a hip fracture during a hypothetical treatment with an effectiveness of 35% targeted to those above the threshold fracture risk. We also examined BMD values in women selected for treatment. Sensitivity analysis examined the effect of age and limited use of BMD resources. RESULTS: BMD tests alone selected women at higher risk of hip fracture than the use of CRFs alone (6.1% versus 5.3%). BMD tests alone also identified a greater number of hip fracture cases (219/1,000) compared to the use of CRFs alone (140/1,000). The combined use of CRFs and BMD identified fewer women above a threshold risk than the use of BMD alone (168/1,000 versus 219/1,000, respectively), but with a higher hip fracture risk (PPV, 8.6% versus 6.1%), and consequently a lower number needed to treat (NNT) (33 versus 47). In sensitivity analyses, the PPV and NNT were always better for the combination than either BMD or CRFs alone across all ages studied (50-70 years). CONCLUSIONS: The use of FRAX in combination with BMD increases the performance characteristics of fracture risk assessment. PMID- 19291346 TI - Abstracts of the Ninth European Congress on Clinical and Economic Aspects of Osteoporosis and Osteoarthritis - ECCEO 9. March 19-21, 2009. Athens, Greece. PMID- 19291350 TI - Use of tumor necrosis factor-alpha (TNF-alpha) antagonists infliximab, etanercept, and adalimumab in patients with concurrent rheumatoid arthritis and hepatitis B or hepatitis C: a retrospective record review of 11 patients. AB - An understanding of the cytokine cascade in a rheumatoid joint has led to the development of new therapeutic options, including drugs targeting tumor necrosis factor-alpha (TNF-alpha). The safety profile of these agents in patients with hepatitis-induced liver disease, however, remains a concern because of risks associated with immune suppression. To examine the effect of three different TNF alpha antagonists, infliximab, etanercept, and adalimumab, on serum transaminases and hepatitis viral load in patients with rheumatoid arthritis (RA) and concurrent hepatitis B (HBV) or hepatitis C (HCV). Medical records of 11 patients with diagnosis of RA and documented seropositivity for hepatitis B or hepatitis C were retrospectively reviewed for worsening of hepatic inflammation and viral proliferation as measured by a rise in aspartate aminotransferase (AST) or alanine aminotransferase (ALT) and viral load while using these agents. Three patients had RA with concurrent chronic HBV and eight patients had RA with concurrent chronic HCV. Seven patients remained on a single anti-TNF-alpha agent and four patients switched to a second anti-TNF-alpha agent due to treatment failure. Two patients showed a transient elevation in AST and/or ALT from normal, but in all 11 patients, AST and ALT levels were within one time the upper range of normal at the conclusion of the study. No significant increase in viral load was seen except one patient who showed a fourfold increase from baseline. Our case series supports results obtained from previous studies examining the safety of anti-TNF-alpha agents in patients with underlying hepatic disease. Use of these agents in patients with HBV or HCV may be associated with a transient transaminitis but appears to be safe overall. In both groups, frequent monitoring of serum transaminase levels and viral load is essential. PMID- 19291351 TI - Anti-chromatin and anti-histone antibodies in Egyptian patients with systemic lupus erythematosus. AB - There has been a renewed interest in anti-chromatin and anti-histone antibodies in the last few years. To assess the prevalence of anti-chromatin and anti histone antibodies in patients with systematic lupus erythematosus (SLE) and to correlate serum levels of these antibodies with clinical features of the disease, the presence of anti-chromatin and anti-histone antibodies in 38 patients with SLE was investigated by an enzyme-linked immunosorbent assay (ELISA). To determine the specificity of these antibodies, 15 patients with rheumatoid arthritis, 15 patients with systemic sclerosis, and 15 normal controls were also tested. Sensitivity of anti-chromatin antibodies in SLE patients was 89.5% and specificity was 80.0%, while sensitivity of anti-histone antibodies was 92.1% and specificity was 82.2%. Significant associations were found between the levels of anti-chromatin antibodies and arthritis, malar rash, oral ulcer, pulmonary affection (P < 0.05) also, lupus nephritis (P < 0.01), and disease activity score as measured by SLE disease activity index (SLEDAI; P < 0.001). Significant association was found between anti-histone antibodies and fatigue (P < 0.05). The incidence of positive anti-chromatin and anti-histone antibodies was significantly higher than that of anti-dsDNA antibodies in early stage of the disease. We conclude that anti-chromatin and anti-histone antibodies are both sensitive and specific for SLE and could be a useful addition to the laboratory tests that can help in the diagnosis of SLE. Anti-chromatin antibodies seem to be a promising marker useful in early diagnosis and assessment of disease activity in SLE patients especially in patients who are negative for anti-dsDNA antibodies. PMID- 19291352 TI - Optical properties of the breast during spontaneous and birth control pill mediated menstrual cycles. AB - Mastodynia is correlated with the menstrual cycle. Using frequency-domain near infrared spectroscopy (FD-NIRS), we investigated changes in breast perfusion in women who were or were not using hormonal contraception. Healthy volunteers, on or not on hormonal contraception, were examined. Optical properties were measured in all quadrants of both breasts, and physiological parameters were calculated. Measurements were repeated every other day during one complete menstrual cycle. Measurements were comparable in all quadrants. Data remained unchanged during the entire cycle in patients using hormonal contraception. However, a biphasic variation of deoxyhemoglobin, oxyhemoglobin, total hemoglobin (tHb), and water content (H(2)O) was observed in women not using contraception. tHb and H(2)O distinctly increased during the ovulation period and remained elevated throughout the luteal phase. It was concluded that FD-NIRS allows accurate measurement of optical properties of human breasts. As opposed to the menstrual cycles of persons using oral contraception, spontaneous menstrual cycles exhibit biphasic variations of tissue perfusion parameters. These findings are important for the investigation of mastodynia. PMID- 19291353 TI - Three synchronous gastrointestinal tumours. AB - Mucinous cystadenomas of the appendix are rare, with significant malignant potential [Lo and Sarr in Hepatogastroenterology 50(50):432-437, 2003]. Carcinoid tumours are similarly uncommon, rarely occurring in Meckel's diverticula but are still the commonest tumour of Meckel's diverticulae [Nies et al. in Dis Colon Rectum 35(6):589-596, 1992; Modlin and Lye in Cancer 97(4):934-959, 2003; Sutter et al. in Schweiz Med Wochenschr Suppl 89:20S-24S, 1997; Weber and McFadden DW in J Clin Gastroenterol 11(6):682-686, 1989]. A 77-year-old woman presented to our clinic with a 6-week history of non-specific lower abdominal pain. A pelvic ultrasound showed an 8 * 3 * 2.5 cm mass in the right iliac fossa. Colonoscopy and CT confirmed this mass and also revealed a left colonic tumor. At laparotomy, three tumours were identified; in the appendix, a Meckel's diverticulum, and the descending colon. A subtotal colectomy and diverticulectomy were performed. Histology confirmed a T3N0 Dukes B colonic adenocarcinoma, a carcinoid of Meckel's diverticulum and a mucinous adenoma of the appendix. The patient is tumor free to date, 5 years after presentation. The presence of three synchronous tumours of different histological origin in the gastrointestinal tract has not previously been described in the literature. PMID- 19291355 TI - Newly developed surface coil for endoluminal MRI, depiction of pig gastric wall layers and vascular architecture in ex vivo study. AB - BACKGROUND: The purpose of this study was to visualize the gastric wall layers and to depict the vascular architecture in vitro by using resected porcine stomachs studied with high-spatial resolution magnetic resonance (MR) imaging. METHODS: Normal dissected porcine stomach samples (n = 4) were examined with a 3 Tesla MR system using a newly developed surface coil. MR images were obtained by the surface coil as receiver and a head coil as transmitter. High-spatial resolution spin-echo MR images were obtained with a field of view of 8 x 8 cm, a matrix of 256 x 128 and slice thicknesses of 3 and 5 mm. RESULTS: T1 and T2 weighted MR images clearly depicted the normal porcine gastric walls as consisting of four distinct layers. In addition, vascular architectures in proper muscle layers were also visualized, which were confirmed by histological examinations to correspond to blood vessels. CONCLUSIONS: High-spatial-resolution MR imaging using a surface coil placed closely to the gastric wall enabled the differentiation of porcine gastric wall layers and the depiction of the blood vessels in proper muscle layer in this experimental study. PMID- 19291354 TI - The role of hedgehog signaling during gastric regeneration. AB - BACKGROUND: Hedgehog signaling plays critical roles during embryonic development. It is also involved in tissue regeneration and carcinogenesis in various adult tissues. Moreover, it regulates the maintenance of cancer stem cells and adult stem cells. Although hedgehog signaling is important in gastric carcinogenesis, its role in gastric regeneration has not been previously examined. In the present study, we evaluated the expression and roles of hedgehog signaling during gastric regeneration. METHODS: Gastric ulcers were induced by serosal application of an acetic acid solution in mice. Sham-operated mice served as controls. The proliferation of gastric progenitor cells was studied using bromodeoxyuridine (BrdU). The expression of hedgehog signaling molecules and the differentiation of gastric progenitor cells were examined by immunohistochemical staining and Western blotting. RESULTS: One day after the induction of gastric ulcer, the proliferation of gastric progenitor cells increased; however, the expression of hedgehog signaling molecules, including sonic hedgehog (Shh), Indian hedgehog (Ihh), desert hedgehog (Dhh), and patched (Ptch1) decreased at the ulcer margin. From 5 days after the induction of gastric ulcer, newly generated gastric glands and their differentiation were observed at the ulcer margin. The expression of hedgehog signaling molecules gradually increased in the newly generated gastric glands of the ulcer margin. Cyclopamine, a specific inhibitor of hedgehog signaling, significantly inhibited the differentiation of mucous cells and parietal cells during the gastric regeneration process. CONCLUSION: The above results suggest that hedgehog signaling is involved in the differentiation of gastric progenitor cells during the gastric ulcer repair process. PMID- 19291356 TI - Standard multivitamin supplementation does not improve vitamin D insufficiency after burns. AB - Children suffering severe burns develop progressive vitamin D deficiency because of inability of burned skin to produce normal quantities of vitamin D(3) and lack of vitamin D supplementation on discharge. Our study was designed to determine whether a daily supplement of a standard multivitamin tablet containing vitamin D(2) 400 IU (10 microg) for 6 months would raise serum levels of 25 hydroxyvitamin D [25(OH)D] to normal. We recruited eight burned children, ages 5 18, whose families were deemed reliable by the research staff. These children were given a daily multivitamin tablet in the hospital for 3 months in the presence of a member of the research staff and then given the remainder at home. At 6 months, the subjects returned for measurements of serum levels of 25(OH)D,1,25-dihydroxyvitamin D [1,25(OH)(2)D], intact parathyroid hormone (iPTH), Ca, P, albumin, and total protein as well as bone mass by dual energy X ray absorptiometry. Serum 25(OH)D levels were compared to a group of seven age matched burned children studied at an earlier date without the vitamin supplement but with the same method of determination of 25(OH)D at 6 months post-burn. In addition, the chewable vitamins were analyzed for vitamin D(2) content by high performance liquid chromatography. Serum concentration of 25(OH)D was 21 +/- 11(SD) ng/ml (sufficient range 30-100) with only one of the eight children having a value in the sufficient range. In comparison, the unsupplemented burn patients had mean serum 25(OH)D level of 16 +/- 7, P = 0.33 versus supplemented. Serum levels of 1,25(OH)(2)D, iPTH, Ca, P, albumin, and total protein were all normal in the supplemented group. Vitamin D(2) content of the chewable tablets after being saponified and extracted was 460 +/- 20 IU. Bone mineral content of the total body and lumbar spine, as well as lumbar spine bone density, failed to increase as expected in the supplemented group. No correlations were found between serum 25(OH)D levels and age, length of stay, percent body surface area burn or third-degree burn. Supplementation of burned children with a standard multivitamin tablet stated to contain 400 IU of vitamin D(2) failed to correct the vitamin D insufficiency. PMID- 19291357 TI - Reticulate hybridization of Alpinia (Zingiberaceae) in Taiwan. AB - Reticulate hybridization is a complicated and creative mechanism in plant evolution that can cause interference in phylogenetic studies. Based on observations of intermediate morphology, low pollen fertility, and overlapping distributions of putative parent species, Yang and Wang (Proceedings of the cross strait symposium on floristic diversity and conservation. National Museum of Natural Science, Taichung, Taiwan, pp 183-197, 1998) first proposed reticulate hybridization of Alpinia in Taiwan. In the present study, molecular tools were used to explore relationships between four parental species and their homoploidy hybrids, and the impact of hybridization on phylogeny reconstruction. Based on DNA markers, maternal heritance of the chloroplast genome, and additivity of nuclear ribosomal internal transcribed spacer, the present results provide strong support for the hybridization hypothesis. Co-existence of parental ribotypes within hybrids revealed that these hybridization events were current, while reciprocal and introgressive hybridization were inferred from chloroplast DNA data. Furthermore, iterative hybridizations involving more than two parental species may occur in notorious hybrid zones. Ecological, phenological, and physiological evidence provides insight into why such frequent hybridization occurs in Taiwanese Alpinia. In the phylogenetic tree of the Zerumbet clade reconstructed in this study, the chloroplast sequences from one hybrid species were not grouped into a subclade, implying instability caused by hybridization. Failure to find morphological apomorphies and biogeographical patterns in this clade was likely partially due to reticulate hybridization. PMID- 19291358 TI - Influence of composition upon the variety of tastes in Cinnamomi cortex. AB - Cinnamomi cortex, which is normally referred to as cinnamon, is a very popular spice as well as an important natural medicine. High-quality cinnamon is traditionally believed to taste sweet and be strongly pungent without astringency. Cinnamomi cortex with larger amounts of cinnamaldehyde was sweeter in taste comparisons. The contents of tannins and sugars in cinnamon powder had little effect on the taste. Evaluations of the sweetness and pungency of cinnamaldehyde solutions (0.1, 0.25, 0.50, 0.75, 1.0, and 5.0 mg/ml) were performed using volunteers. The scores for sweetness increased significantly from 0.10 to 0.50 mg/ml (P < 0.05, Mann-Whitney U-test), but there was no significant difference above 0.75 mg/ml. The concentration threshold for the sweet taste of cinnamaldehyde appeared to be less than 0.75 mg/ml, and the more concentrated solutions gave excessive pungency. Therefore, two contrastive tastes of Cinnamomi cortex, sweet and pungent, were both attributed to cinnamaldehyde. Consequently, its taste, one of its indices of quality, seems to vary mainly according to the content of cinnamaldehyde. PMID- 19291359 TI - Survival after cord blood transplantation from unrelated donor as a second hematopoietic stem cell transplantation for recurrent pediatric acute myeloid leukemia. AB - The Japan Cord Blood Bank Network (JCBBN) reports the treatment of 22 children with acute myeloid leukemia (AML) who received umbilical cord blood transplantation from unrelated donors (CBT) as their second hematopoietic stem cell transplantation (HSCT). Provided by the JCBBN, between February 1997 and September 2006, 22 patients had CBT as a second HSCT. In the initial HSCT, eight received autologous, seven received CBT, and the remaining had allogenic BMT. At the time of CBT as a second HSCT, seven were in the second complete remission (CR2), two in the third CR (CR3), the remaining were not in remission. Reduced intensity conditioning (RIC) conducted for 10 cases and myeloablative conditioning (MAC) for 12 cases. The overall survival rate was 31.3%, 5 years after CBT. Second complete remission at second transplantation was favorable prognosis (58.3 +/- 18.6%, compared with 17.1 +/- 10.8% for the non-CR group. Mortality after CBT as a second HSCT accounted for 15 cases, 8 from treatment related mortality. In conclusion, CBT combined with RIC as second HSCT may be useful against a recurrence of AML in children after the initial HSCT. PMID- 19291361 TI - The nucleotide sequences of the RNA 1 and RNA 2 of asparagus virus 2 show a close relationship to citrus variegation virus. PMID- 19291362 TI - Herpes simplex virus type 1 infection increases the carbohydrate binding activity and the secretion of cellular galectin-3. AB - Galectin-3 binds beta-galactoside-containing sugars and is a chemoattractant for monocytes, macrophages, and neutrophils. Galectin-3 was identified by mass spectrometry from an anti-gI affinity column; however, we determined that galectin-3 did not bind gI, but rather that HSV-1 infection increased galectin-3 binding to carbohydrate residues on IgG. Our conclusions are based on the following observations: (1) galectin-3 from cells infected with a gI-deleted HSV 1 mutant virus bound anti-gI IgG; (2) galectin-3 from wild-type HSV-1 infected cells bound nonimmune IgG; (3) more galectin-3 from infected than uninfected cells bound IgG; and (4) binding to IgG was blocked by lactose, a competitive inhibitor of galectin-3 carbohydrate binding. HSV-1 infection did not increase galectin-3 expression, but did increase its secretion. We propose that increased carbohydrate binding and secretion of galectin-3 contribute to an early pro inflammatory innate immune response to HSV-1 infection. PMID- 19291360 TI - The FGF-2/FGFRs neurotrophic system promotes neurogenesis in the adult brain. AB - Neurogenesis occurs in two regions of the adult brain, namely, the subventricular zone (SVZ) throughout the wall of the lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus (DG) in hippocampal formation. Adult neurogenesis requires several neurotrophic factors to sustain and regulate the proliferation and differentiation of the adult stem cell population. In the present review, we examine the cellular and functional aspects of a trophic system mediated by fibroblast growth factor-2 (FGF-2) and its receptors (FGFRs) related to neurogenesis in the SVZ and SGZ of the adult rat brain. In the SVZ, FGF-2 is expressed in GFAP-positive cells of SVZ but is not present in proliferating precursor cells, which instead express FGFR-1 and FGFR-2, but not FGFR-3 mRNA, although expressed in the SVZ, and FGFR-4. Therefore, it seems that in the SVZ FGF-2 may be released by GFAP-positive cells, different from the precursor cell lineage, and via volume transmission it reaches the proliferating precursor cells. FGFR-1 mRNA is also expressed in the SGZ and is localized in BrdU-labeled precursor cells, whereas FGFR-2 and FGFR-3 mRNA, although expressed in the SGZ, are not located within proliferating precursor cells. An aged-related decline of proliferating precursor cells in the SVZ and DG of old rats has been well documented, and there is the suggestion that in part it could be the consequence of alterations in growth factor expression levels. Thus, the old precursors may respond to growth factors, suggesting that during aging the basic components for neuronal precursor cell proliferation are retained and the capacity to increase neurogenesis after appropriate stimulation is still preserved. In conclusion, the trophic system mediated by FGF-2 and its receptors contributes to create an important micro-environmental niche that promotes neurogenesis in the adult and aged brain. PMID- 19291363 TI - Antiviral activity of Arbidol against Coxsackie virus B5 in vitro and in vivo. AB - We investigated the antiviral activity of Arbidol, an antiviral chemical compound, against Coxsackie virus B5 (CVB(5)) in vitro and in vivo. Arbidol not only prevented the cytopathic effect (CPE) of CVB(5), as demonstrated in an MTT colorimetric assay, when added during or after viral infection, with a 50% inhibitory concentration (IC(50)) from 2.66 to 6.62 microg/ml, but it also decreased the CVB(5)-RNA level in infected host cells, as shown in semi quantitative RT-PCR. BALB/c mice were used as an animal model to test the Arbidol activity in vivo. Orally administered Arbidol at 50 mg/kg body weight/day (once a day) significantly reduced mean virus yields in the lungs and heart as well as mortality after infection for 6 days. Our results demonstrate that in vitro and in vivo infection with CVB(5) can be effectively treated by Arbidol. PMID- 19291364 TI - Damage in needle tissues after infection with Chrysomyxa rhododendri increases cuticular conductance of Picea abies in winter. AB - Chrysomyxa rhododendri is a rust which infects Picea abies growing near the alpine timberline. Attacked needles are normally shed, but few remain on shoots. We hypothesised that these needles increase transpiration of Picea during winter. Partly damaged, completely damaged and healthy needles of an infected tree as well as healthy needles of a resistant tree were compared in a microscopy analysis, and needle conductance of shoots was measured gravimetrically. Despite needle shedding, more than 6% of needles remaining on infected tree shoots were damaged. Partly damaged needles showed local brownish areas in the periphery and completely damaged needles necrotic parenchyma and epidermal tissues. Cuticular conductance of affected shoots was up to 25.23 +/- 2.75 mmol m(-2) s(-1) at moderate water potential and thus twofold higher than in the resistant tree. Needle shedding reduces negative effects of Chrysomyxa infections during summer, but remaining damaged needles impair tree water relations in winter. PMID- 19291365 TI - Catabolism of nutritionally essential amino acids in developing porcine enterocytes. AB - This study was conducted using the piglet model to test the hypothesis that mucosal cells of the neonatal small intestine can degrade nutritionally essential amino acids (EAA). Enterocytes were isolated from the jejunum of 0-, 7-, 14-, and 21-day-old pigs, and incubated for 45 min in Krebs buffer containing plasma concentrations of amino acids and one of the following L-[1-(14)C]- or L-[U (14)C]-amino acids plus unlabeled tracees at 0.5, 2, or 5 mM: histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine. In these cells, branched-chain amino acids (BCAA) were extensively transaminated and 15-50% of decarboxylated branched-chain alpha-ketoacids (BCKA) were oxidized to CO(2) depending on the age of piglets. BCAA transamination increased but their decarboxylation decreased between 0 and 14 days of age. Addition of 1 and 2 mM alpha-ketoglutarate to incubation medium dose-dependently stimulated BCAA transamination without affecting their decarboxylation. Western blot analysis revealed that the abundance of mitochondrial BCAA aminotransferase declined but cytosolic BCAA aminotransferase increased between 0 and 14 days of age, with the cytosolic protein being the major isoform in 7- to 21-day-old pigs. BCKA dehydrogenase protein existed primarily as the phosphorylated (inactive) form in enterocytes of newborn pigs and its levels were markedly reduced in older pigs. All measured parameters of BCAA metabolism did not differ between 14- and 21-day-old pigs. In contrast to BCAA, catabolism of methionine and phenylalanine was negligible and that of other EAA was absent in enterocytes from all ages of piglets due to the lack of key enzymes. These results indicate that enterocytes are an important site for substantial degradation of BCAA but not other EAA in the neonatal gut. PMID- 19291366 TI - Tumor-targeting amino acid auxotrophic Salmonella typhimurium. AB - We have developed an effective bacterial cancer therapy strategy by targeting viable tumor tissue using Salmonella typhimurium auxotrophs that we have generated which grow in viable as well as necrotic areas of tumors. However, the auxotrophy severely restricts growth of these bacteria in normal tissue. The S. typhimurium A1-R mutant, which is auxotrophic for leu-arg and has high anti-tumor virulence, was developed in our laboratory. In vitro, A1-R infects tumor cells and causes nuclear destruction. A1-R was initially used to treat metastatic human prostate and breast tumors that had been orthotopically implanted in nude mice. Forty percent of treated mice were cured completely and survived as long as non tumor-bearing mice. A1-R administered i.v. to nude mice with primary osteosarcoma and lung metastasis was highly effective, especially against metastasis. A1-R was also targeted to both axillary lymph and popliteal lymph node metastasis of human pancreatic cancer and fibrosarcoma, respectively, as well as lung metastasis of the fibrosarcoma in nude mice. The bacteria were delivered via a lymphatic channel to target the lymph node metastases and systemically via the tail vein to target the lung metastasis. The metastases were cured without the need of chemotherapy or any other treatment. A1-R was administered intratumorally to nude mice with an orthotopically transplanted human pancreatic tumor. The primary pancreatic cancer regressed without additional chemotherapy or any other treatment. A1-R was also effective against pancreatic cancer liver metastasis when administered intrasplenically to nude mice. The approach described here, where bacterial monotherapy effectively treats primary and metastatic tumors, is a significant improvement over previous bacterial tumor-therapy strategies that require combination with toxic chemotherapy. Three promoter clones engineered in S. enterica typhimurium were identified to have enhanced expression in bacteria growing in tumors relative to those growing in the spleen. The expression of therapeutics in Salmonella under the regulation of one or more promoters that are activated preferentially in tumors has the potential to improve the efficacy of Salmonella tumor therapy. Exploitation of the tumor-killing capability of Salmonella has great promise for a new paradigm of cancer therapy. PMID- 19291367 TI - The pharmacodynamics of enoxaparin in percutaneous coronary intervention with precise rapid enoxaparin loading (PEPCI-PRE study). AB - OBJECTIVE: We evaluated the early pharmacodynamic profile of the combined 30 mg intravenous and 1 mg/kg subcutaneous enoxaparin loading utilized in the TIMI 11B and ExTRACT TIMI 25 trials. BACKGROUND: It has not been reported whether anti-Xa levels appropriate for percutaneous coronary intervention (PCI) can be reliably achieved within 2 h utilizing this regimen. METHODS: Twenty-six patients with acute coronary syndrome (ACS) treated with this regimen had anti-Xa levels measured at 5 min, 2, 4, 6 and 8 h. RESULTS: Seventy-six percent of patients had anti-Xa levels above 0.5 IU/ml at 5 min. Dose-response curves showed all patients to have anti-Xa levels above 0.5 IU/ml within 1 h. Anti-Xa remained in the targeted range for PCI (0.5 to 1.8 IU/ml) at 2, 4, 6 and 8 h in all patients. CONCLUSION: This regimen is well suited for ACS treatment with an invasive strategy, including the rapid transition to early and rescue PCI. PMID- 19291368 TI - The early osseointegration of the laser-treated and acid-etched dental implants surface: an experimental study in rabbits. AB - The aim of the study was to evaluate early osseointegration of the laser-treated and acid-etched implant surface after the installation in rabbit tibias for 4 weeks. A total of 56 screw-shaped implants were grouped as follows: group A: implants were turned surface; group B: implants were laser-treated surface; group C: implants were acid-etched; group D: Implants were laser-treated and acid etched surface. After 4 weeks, the removal torques were: group A: 13.21 +/- 11.30 Ncm; group B: 29.73 +/- 8.32 Ncm; group C: 30.31 +/- 9.45 Ncm; group D: 35.76 +/- 7.58 Ncm; The averages of bone-to-implant contact (BIC) were as follows: group A: 27.30 +/- 6.55%; group B: 38.00 +/- 8.56%; group C: 42.71 +/- 8.48%; group D: 49.71 +/- 9.21%. The removal torque and bone-to-implant contact measurements yielded statistically significant differences between the treated groups and turned group (P < 0.05); The laser-treated and acid-etched surface achieved higher Bone-to-Implant Contact than the laser-treated surface (P < 0.05), but there was no statistically significant difference between the laser-treated and acid-etched surface and the acid-etched surface in bone-to-implant contact (P > 0.05). In the present study, it was concluded that the laser-treated and acid etched implants had good osteoconductivity and was a potential material for dental implantation. PMID- 19291369 TI - Addition of sodium hyaluronate and the effect on performance of the injectable calcium phosphate cement. AB - An injectable calcium phosphate cement (CPC) with porous structure and excellent anti-washout ability was developed in the study. Citric acid and sodium bicarbonate were added into the CPC powder consisting of tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD) to form macro-pores, then different concentrations of sodium hyaluronate (NaHA) solution, as liquid phase, was added into the cement to investigate its effect on CPC's performance. The prepared CPCs were tested on workability (injectable time and setting time), mechanical strength, as well as anti-washout ability. The experimental results showed that addition of NaHA not only enhanced the anti-washout ability of the CPC dramatically but also improve its other properties. When NaHA concentration was 0.6 wt%, the injectable time elongated to 15.7 +/- 0.6 min, the initial and final setting times were respectively shorten to 18.3 +/- 1.2 and 58.7 +/- 2.1 min, and the compressive strength were increased to 18.78 +/- 1.83 MPa. On the other hand, Addition of NaHA showed little effect on porous structure of the CPC and enhanced its bioactivity obviously, which was confirmed by the apatite formation on its surface after immersion in simulated body fluid (SBF). In conclusion, as an in situ shaped injectable biomaterials, the CPC with appropriate addition of NaHA would notably improve its performance and might be used in minimal invasive surgery for bone repair or reconstruction. PMID- 19291371 TI - Transferrin receptor targeted lipopolyplexes for delivery of antisense oligonucleotide g3139 in a murine k562 xenograft model. AB - PURPOSE: Transferrin (Tf) conjugated lipopolyplexes (LPs) carrying G3139, an antisense oligonucleotide for Bcl-2, were synthesized and evaluated in Tf receptor positive K562 erythroleukemia cells and then in a murine K562 xenograft model. MATERIALS AND METHODS: Particle size and Zeta potentials of transferrin conjugated lipopolyplexs containing G3139 (Tf-LP-G3139) were measured by Dynamic Light Scattering and ZetaPALS. In vitro and in vivo sample's Bcl-2 downregulation was analyzed using Western blot and tumor tissue samples also exhibited by immunohistochemistry method. For athymic mice bearing with K562 xenograft tumors, tumor growth inhibition and survival rate were investigated. Nanoparticle distribution in 3-D cell cluster was observed by Laser scan confocal microscopy. IL-12 production in the plasma was measured by ELISA kit. RESULTS: In vitro, Tf LP-G3139 was more effective in inducing down regulation of Bcl-2 in K562 cells than non-targeted LP-G3139, free G3139 and mismatched control ODN-G4126 in the same formulation. In vivo Tf-LP-G3139 was less effective than free G3139 in Bcl-2 down regulation. 3-D cell cluster model diffusion results indeed indicated limited penetration of the LPs into the cell cluster. Finally, the therapeutic efficacies of Tf-LP-G3139 and free G3139 were determined in the K562 xenograft model. Tf-LP-G3139 showed slower plasma clearance, higher AUC, and greater accumulation in the tumor compared to free G3139. In addition, Tf-LP-G3139 was found to be more effective in tumor growth inhibition and prolonging mouse survival than free G3139. This was associated with increased spleen weight and IL 12 production in the plasma. CONCLUSION: The role of the immune system in the therapeutic response obtained with the Tf-LPs is necessary and in vitro 3-D cell cluster model can be a potential tool to evaluate the nanoparticle distribution. PMID- 19291372 TI - A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels. AB - PURPOSE: To simultaneously quantify intracellular nucleoside triphosphate (NTP) and deoxynucleoside triphosphate (dNTP) pools and to assess their changes produced by interfering with ribonucleotide reductase (RNR) expression in leukemia cells. METHODS: A HPLC-MS/MS system was used to quantify intracellular NTP and dNTP pools. RESULTS: The assay was linear between 50 nM, the lower limit of quantification (LLOQ), and 10 muM in cell lysate. The within-day coefficients of variation (CVs, n = 5) were found to be 12.0-18.0% at the LLOQ and 3.0-9.0% between 500 and 5,000 nM for dNTPs and 8.0-15.0% and 2.0-6.0% for NTPs. The between-day CVs (n = 5) were 9.0-13.0% and 3.0-11.0% for dNTPs and 9.0-13.0% and 3.0-6.0% for NTPs. The within-day accuracy values were 93.0-119.0% for both NTPs and dNTPs. ATP overlapped with dGTP and they were analyzed as a composite. This method was applied to measure basal intracellular dNTPs/NTPs in five leukemia cell lines exposed to the RNR antisense GTI-2040. Following drug treatment, dCTP and dATP levels were found to decrease significantly in MV4-11 and K562 cells. Additionally, perturbation of dNTP/NTP levels in bone marrow sample of a patient treated with GTI-2040 was detected. CONCLUSIONS: This method provides a practical tool to measure intracellular dNTP/NTP levels in cells and clinical samples. PMID- 19291373 TI - Role of transporters in ocular drug delivery system. PMID- 19291374 TI - ICOS deficiency results in exacerbated IL-17 mediated experimental autoimmune encephalomyelitis. AB - INTRODUCTION: Inducible costimulatory molecule (ICOS) is important for the effector function of T cells, especially for Th2 and T cell dependent B cell responses. However, it has been shown that ICOS is required for the differentiation of Th17 cells. Since IL-17 has been identified as a major cytokine involved in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), the enhanced severity of EAE in ICOS-deficient mice (ICOS(-/-)) mice is unexpected. METHODS: To better understand the role of ICOS and of IL-17 in EAE, we induced EAE in ICOS(-/-) by immunization with myelin oligodendrocyte glycoprotein peptide (MOG(35-55)) in complete Freund's adjuvant. RESULTS: As previously reported, we found that ICOS(-/-) mice developed more severe EAE. Upon restimulation with MOG(35-55,) splenocytes from ICOS(-/-) mice with EAE produced higher amounts of IL-17 and ICOS(-/-) mice had a higher expression of IL-17, IL-6, and TGF-beta mRNA in the spinal cords at the onset of the disease. Finally, the blockade of IL-17 strongly inhibited disease even in ICOS(-/-) mice, showing that IL-17 is playing a major role in the pathogenesis of EAE both in WT and ICOS(-/-) mice. CONCLUSION: In conclusion, MOG immunization induces MOG-specific Th17 cells also in ICOS(-/-) mice, and a higher expression of IL-17 and of Th17-driving cytokines IL-6 and TGF-beta in the central nervous system at the onset of EAE that correlates with their more severe disease. PMID- 19291375 TI - Imbalances between interleukin-1 and tumor necrosis factor agonists and antagonists in stable COPD. AB - INTRODUCTION: Interleukin-1 beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) are potentially important in Chronic Obstructive Pulmonary Disease (COPD), but little is known of the relationships between these cytokines and their antagonists in disease compared with healthy controls. It is unclear if concentrations relate to disease severity. The study aimed to investigate these relationships and to assess the potential activity of each cytokine in the context of their antagonists. METHODS: Plasma cytokines, soluble receptors, and cell counts were measured in patients with stable COPD and age-matched healthy controls (n = 15 for both) daily for 5 days; these mediators were also measured in corresponding sputum samples from the COPD patients. RESULTS: COPD patients had significantly reduced concentrations of the antagonists, IL-1sRII, and IL-1RA compared with controls. In COPD, IL-1beta exceeded its antagonists and correlated significantly with BMI and FEV1, while plasma IL-1RA correlated positively with BMI but negatively with sputum IL-1beta, neutrophil, and macrophage counts and smoking history. TNFalpha antagonists exceeded agonists in both groups and did not correlate with COPD severity. CONCLUSIONS: Endogenous IL-1beta antagonists appear reduced in COPD. Furthermore, IL-1beta correlated with clinical aspects of disease severity, suggesting that IL-1beta may play a critical role in COPD. Given the relevant concentrations and binding affinities, it is likely that TNFalpha has limited activity in stable COPD. PMID- 19291376 TI - A systematic review of the impact of genetic counseling on risk perception accuracy. AB - This review presents an overview of the impact of genetic counseling on risk perception accuracy in papers published between January 2000 and February 2007. The results suggest that genetic counseling may have a positive impact on risk perception accuracy, though some studies observed no impact at all, or only for low-risk participants. Several implications for future research can be deduced. First, future researchers should link risk perception changes to objective risk estimates, define risk perception accuracy as the correct counseled risk estimate, and report both the proportion of individuals who correctly estimate their risk and the average overestimation of the risk. Second, as the descriptions of the counseling sessions were generally poor, future research should include more detailed description of these sessions and link their content to risk perception outcomes to allow interpretation of the results. Finally, the effect of genetic counseling should be examined for a wider variety of hereditary conditions. Genetic counselors should provide the necessary context in which counselees can understand risk information, use both verbal and numerical risk estimates to communicate personal risk information, and use visual aids when communicating numerical risk information. PMID- 19291377 TI - The interaction of positive and negative sensory feedback loops in dynamic regulation of a motor pattern. AB - In many rhythmic behaviors, phasic sensory feedback modifies the motor pattern. This modification is assumed to depend on feedback sign (positive vs. negative). While on a phenomenological level feedback sign is well defined, many sensory pathways also process antagonistic, and possibly contradictory, sensory information. We here model the locust flight pattern generator and proprioceptive feedback provided by the tegula wing receptor to test the functional significance of sensory pathways processing antagonistic information. We demonstrate that the tegula provides delayed positive feedback via interneuron 301, while all other pathways provide negative feedback. Contradictory to previous assumptions, the increase of wing beat frequency when the tegula is activated during flight is due to the positive feedback. By use of an abstract model we reveal that the regulation of motor pattern frequency by sensory feedback critically depends on the interaction of positive and negative feedback, and thus on the weighting of antagonistic pathways. PMID- 19291378 TI - Brief report: Effects of pressure vest usage on engagement and problem behaviors of a young child with developmental delays. AB - The purpose of this study was to examine the effects of wearing a pressure vest for a young boy with developmental delays. An A-B-A withdrawal design was used to examine the relation between wearing the pressure vest and child behaviors during a preschool art activity. Although the data showed moderate variability, no systematic differences were found in child engagement when the vest was worn and when the vest was not worn and problem behavior increased when the vest was being worn. These results are discussed in the context of the study limitations. Implications for future research are provided. PMID- 19291379 TI - Adolescent siblings of individuals with an autism spectrum disorder: testing a diathesis-stress model of sibling well-being. AB - The purpose of this study was to test a diathesis-stress model of well-being for siblings who have a brother or sister with an autism spectrum disorder (ASD). Data were collected from 57 adolescents and their mothers. Sisters reported higher levels of depressive and anxiety symptoms than brothers. Having a family history of ASDs was associated with depressive, but not anxiety, symptoms. A high level of maternal depression was also associated with more depressive and anxiety symptoms. A diathesis-stress model was partially supported, primarily through the findings that sibling sub-threshold autism characteristics were associated with depressive and anxiety symptoms in siblings, but only in the presence of a high number of stressful life events. PMID- 19291380 TI - Inhibition and the validity of the Stroop task for children with autism. AB - Findings are mixed concerning inhibition in autism. Using the classic Stroop, children with autism (CWA) often outperform typically developing children (TDC). A classic Stroop and a chimeric animal Stroop were used to explore the validity of the Stroop task as a test of inhibition for CWA. During the classic Stroop, children ignored the word and named the ink colour, then vice versa. Although CWA showed less interference than TDC when colour naming, both groups showed comparable interference when word reading. During the chimeric animal task, children ignored bodies of animals and named heads, and vice versa; the groups performed comparably. Findings confirm that lower reading comprehension affects Stroop interference in CWA, potentially leading to inaccurate conclusions concerning inhibition in CWA. PMID- 19291381 TI - Autism and dimensionality: differences between copying and drawing tasks. AB - Previous research suggests individuals with autism may be less influenced by a three-dimensional interpretation when copying line drawings (Sheppard et al. J Autism Dev Disord 37:1913-1924, 2007). The current research aimed to determine whether this reduced dimensionality effect extends to drawings of an actual object. Twenty-four children and adolescents with autism and 24 comparison participants copied one line drawing with no depth cues, line drawings with a three-dimensional interpretation, and drew a actual three-dimensional object. Participants with autism were less influenced by three-dimensionality on the copying tasks but were equally affected when drawing the actual object. This suggests that any advantage for three-dimensional drawing in non-savant individuals with autism is confined to situations when the individual copies a line drawing with depth cues. PMID- 19291382 TI - Prenatal stress and risk of febrile seizures in children: a nationwide longitudinal study in Denmark. AB - We aimed to examine whether exposure to prenatal stress following maternal bereavement is associated with an increased risk of febrile seizures. In a longitudinal population-based cohort study, we followed 1,431,175 children born in Denmark. A total of 34,777 children were born to women who lost a close relative during pregnancy or within 1 year before the pregnancy and they were included in the exposed group. The exposed children had a risk of febrile seizures similar to that of the unexposed children (hazard ratio (HR) 1.00, 95% CI 0.94-1.06). The HRs did not differ according to the nature or timing of bereavement. Our data do not suggest any causal link between exposure to prenatal stress and febrile seizures in childhood. PMID- 19291383 TI - Supervisor support as a predictor of burnout and therapeutic self-efficacy in therapists working in ABA schools. AB - Very little is known about factors potentially affecting the performance of therapists delivering applied behavior analysis (ABA) interventions for young children with autism. Eighty-one therapists working in ABA schools participated in a questionnaire study focused on their reports of burnout and perceived therapeutic self-efficacy in their work role. Perceived supervisor support played a central role in the prediction of reduced therapist burnout and increased therapeutic self-efficacy. In addition, perceived supervisor support moderated the impact of work demands on personal accomplishment burnout. Those therapists reporting high work demands and lower levels of supervisor support had lower personal accomplishment scores on the Maslach burnout inventory. Clinical implications include the importance of supervisor support for therapists and also supervisor style. PMID- 19291384 TI - Cytomegalovirus microRNAs. AB - MicroRNAs (miRNAs) are small, non-coding RNA molecules that regulate gene expression at a post-transcriptional level in virtually all eukaryotic organisms and some viruses, particularly herpesviruses. miRNAs are non-immunogenic, stealthy tools for viruses to regulate their as well as host gene expression. The human cytomegalovirus (HCMV) is the major cause of morbidity in immunocompromised patients and allogenic bone-marrow or organ-transplant recipients and the leading cause of congenital birth defects. HCMV miRNAs may provide valuable targets for new urgently needed antiviral drugs. This review focuses on recent findings for viral miRNAs expressed by cytomegaloviruses (CMV) including data from human, chimpanzee, and murine CMV. These are discussed in the context of findings for other viruses to highlight potentially conserved roles exerted by viral miRNAs. PMID- 19291385 TI - Parents, peers, and sexual values influence sexual behavior during the transition to college. AB - Several decades of research have identified the contributions of psychosocial influences on adolescent and young adult sexual behavior; however, few studies have examined parental and peer influence and sexual values during the transition from high school to college. The current study tested the influence of sexual values and perceived awareness and caring (PAC), or beliefs about how much parents and peers know and care about students' behavior, on sexual behavior during this transitional period. Using data from a longitudinal study, generalized estimating equations and the generalized linear model were used to examine the associations among sexual values, parental and peer PAC, and sexual behavior, both cross-sectionally and longitudinally. Participants (N = 1,847; 61% female) completed web-based surveys the summer before college matriculation and at the end of the first semester in college. Results indicated that individuals with high levels of both parental and peer PAC engaged in less frequent sexual behaviors and that PAC moderated the effect of sexual values on sexual behaviors. Furthermore, both PAC variables decreased during the transition from high school to college, and high school sexual values, parental PAC, and their interaction predicted the number of sexual partners during the first semester of college. Only sexual values and high school unsafe sexual behaviors predicted unsafe sexual behavior in college. Findings suggest that complex associations exist among perceived awareness and caring, sexual values, and sexual behaviors, and that the transition from high school to college may be an ideal time for safer sex interventions. PMID- 19291386 TI - The relationship between neuropsychological functioning and HAART adherence in HIV-positive adults: a systematic review. AB - Combination antiretroviral therapy has helped extend the lives of persons infected with HIV; however, the efficacy of highly active antiretroviral therapy (HAART) regimens depends, in part, on the consistency with which the medications are taken. In this paper, we review 11 empirical studies conducted in Western developed nations that utilized psychometrically valid neuropsychological measures to examine the relationship between cognitive functioning and HAART adherence. In general, impaired neuropsychological functioning--particularly within the domains of executive functioning and problem solving, learning and memory, attention and working memory, and global cognitive functioning--was associated with lower medication adherence across studies. However, inconsistent operationalizations of neuropsychological impairment and medication adherence employed in these studies, as well as the paucity of longitudinal data to support temporal relationships, may attenuate these conclusions. We conclude with a set of research recommendations that may help to improve the rigor of future studies and clarify questions left unanswered due to methodological limitations of existing studies. PMID- 19291387 TI - Stimulant treatment reduces lapses in attention among children with ADHD: the effects of methylphenidate on intra-individual response time distributions. AB - Recent research has suggested that intra-individual variability in reaction time (RT) distributions of children with ADHD is characterized by a particularly large rightward skew that may reflect lapses in attention. The purpose of the study was to provide the first randomized, placebo-controlled test of the effects of the stimulant methylphenidate (MPH) on this tail and other RT distribution characteristics. Participants were 49 9- to 12-year-old children with ADHD. Children participated in a 3-day double-blind, placebo-controlled medication assessment during which they received long-acting MPH (Concerta), with the nearest equivalents of 0.3 and 0.6 mg/kg t.i.d. immediate-release MPH. Children completed a simple two-choice speeded discrimination task on and off of medication. Mode RT and deviation from the mode were used to examine the peak and skew, respectively, of RT distributions. MPH significantly reduced the peak and skew of RT distributions. Importantly, the two medication effects were uncorrelated suggesting that MPH works to improve both the speed and variability in responding. The improvement in variability with stimulant treatment is interpreted as a reduction in lapses in attention. This, in turn, may reflect stimulant enhancement of self-regulatory processes theorized to be at the core of ADHD. PMID- 19291388 TI - A test of a cognitive diathesis-stress generation pathway in early adolescent depression. AB - This study evaluates a pathway for depressive risk that integrates cognitive diathesis-stress and stress-generation theories, following Hankin and Abramson's (2001, Journal of Clinical Child and Adolescent Psychology, 31(4), 491-504) elaborated cognitive-diathesis transactional stress model. In this model, young adolescents with initial depressive symptoms were hypothesised to experience later stressors that were at least partly dependent on their behaviour. The interaction of cognitive vulnerability, a tendency to make depressogenic attributions and to ruminate, with these dependent stressors was then hypothesised to predict depressive symptoms after 6 months. This model was supported in a sample of 756 young adolescents, with cognitive style and dependent stressors partly mediating the relationship between initial and subsequent depressive symptoms. Cognitive vulnerability was also linked with an increased likelihood of dependent stressors. PMID- 19291389 TI - Novel mutations of the FASN gene and their effect on fatty acid composition in Japanese Black beef. AB - Eight novel and four known mutations were detected in the coding sequence of the bovine fatty acid synthase (FASN) gene of an F2 population from Japanese Black and Limousin cattle. Two mutations, g.16024A>G and g.16039T>C, detected in exon 34, which determine amino acid substitutions of threonine (T) to alanine (A) and tryptophan (W) to arginine (R), were clearly separated in the parental breeds. The haplotypes (TW and AR) segregated in F2 individuals and had a significant effect on the fatty acid composition of backfat, intermuscular fat, and intramuscular fat. The TW haplotype was associated with increasing C18:0 and C18:1 content and the ratio of monounsaturated to saturated fatty acids, and decreasing C14:0, C14:1, C16:0, and C16:1 content. The two mutations were screened in two commercial Japanese Black half-sibling populations and similarly determined the contribution to the fatty acid composition of intramuscular fat. The frequency of the TW haplotype was markedly higher in Japanese Black (0.67) than in Holstein (0.17), Angus (0.02), and Hereford (0.07). We conclude that these mutations may contribute to the characteristic fatty acid composition of Japanese Black beef. PMID- 19291390 TI - Alpha2-adrenergic impact on hypothalamic magnocellular oxytocinergic neurons in long evans and brattleboro rats: effects of agonist and antagonists. AB - We have previously demonstrated that alpha2-adrenoceptors regulate hypothalamic magnocellular oxytocinergic (OXY) neurons in Sprague Dawley rats. Here we investigated whether activation/inhibition of alpha2-adrenoceptors may similarly trigger/downregulate the activity of OXY neurons in control Long Evans (+/+) and permanently osmotically stressed Brattleboro (di/di) rats. The effect of alpha2 adrenoceptor agonist, xylazine (XYL) and alpha2-adrenoceptor antagonists, atipamezole (ATIP), and idazoxan (IDX) were evaluated in the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. Saline (SAL, 0.1 ml/100 g), XYL (10 mg/kg), ATIP, (1 mg/kg), and IDX (10 mg/kg) and IDX or ATIP followed by XYL were applied intraperitoneally. Rats were sacrificed 90 min later and Fos/OXY co labelings analyzed in microscope. In control +/+ rats no or few Fos/OXY co labelings occurred in SON and PVN. XYL significantly increased Fos incidence in OXY neurons in both nuclei. ATIP significantly suppressed the effect of XYL in both nuclei and IDX only in SON. In di/di controls 81% of OXY neurons in SON and 44% in PVN revealed Fos presence and XYL did not further elevate Fos number in SON OXY neurons and slightly increased Fos number in PVN. ATIP or IDX only partially reduced Fos in SAL or XYL treated di/di rats. Our data indicate that: (1) XYL stimulation is not effective in di/di rats because of sustained upregulation of OXY neurons activity and (2) neither ATIP nor IDX reduced significantly the OXY activity in control di/di rats. These findings suggest that alpha2-adrenoceptors have only a limited impact in maintaining OXY cells activity upregulation in PVN and SON of di/di rats. PMID- 19291391 TI - Effect of noradrenalin and EGb 761 pretreatment on the ischemia-reperfusion injured spinal cord neurons in rabbits. AB - Short term sublethal ischemia or ischemic preconditioning gives protection to the neurons against subsequent lethal ischemic attack. This so-called ischemic tolerance can also be provided by certain drugs. We examined the effect of noradrenalin and EGb 761 on the spinal cord neurons injured by 30 min occlusion of abdominal aorta in rabbits. The animals survived 48 and 72 h. Degenerated neurons were visualized by Fluoro Jade B method, viable neurons were demonstrated immunohistochemically with NeuN and ubiquitin antibodies. The rabbits with noradrenalin administration 48 h before 30 min of ischemia and 48/72 h of reperfusion, showed significant increase of degenerated Fluoro Jade B labeled neurons. Animals of both groups were paraplegic. Rabbits pretreated 7 days with EGb 761 prior to 30 min of ischemia and with 48/72 h of reperfusion revealed significant decrease of Fluoro Jade B-positive neurons when compared with the groups with 30 min of ischemia followed by 48/72 h of reperfusion. In the NeuN sections, the number of viable neurons was moderately decreased. These animals showed no paraplegia. Ubiquitin aggregates occurred in the cytoplasm of degenerated neurons in the sections of rabbits preconditioned with noradrenalin 48 h prior to 30 min of ischemia and followed by 48 h of reperfusion while after 72 h of reperfusion, shrunk light shadows without ubiquitin reaction were visible. Our results indicate that EGb 761 could be involved in protection of spinal cord neurons against ischemic injury while effect of noradrenalin is not unambiguous. PMID- 19291392 TI - Event-related desynchronization/synchronization during discrimination task conditions in patients with Parkinson's disease. AB - Parkinson's disease is a neurodegenerative disorder with symptoms, which include movement disturbances and changes of cognitive information processing. The aim of the present study was to investigate the functional relationships between oscillatory electroencephalographic (EEG) dominant components with event-related desynchronization/synchronization (ERD/ERS) method for idiopathic non-demented Parkinson's patients (PP) and control subjects (CS) during auditory discrimination tasks within two post-stimulus intervals of 0-250 and 250-600 ms. When comparing the CS and PP during the first post-stimulus period, we found delta- and theta-ERS significantly pronounced in CS for both tone types (low- 800, high--1,000 Hz) with the following exceptions: at Fz, PP displayed higher delta-ERS, while at C3' theta-ERD in response to a high tone. Alpha-ERS was found in PP in response to either tone at all electrodes and mainly alpha-ERD in CS. In the second post-stimulus interval, the significant differences between the groups were: (i) delta-ERS in CS and delta-ERD in PP in response to the low tone and (ii) delta-ERS for both groups in answer to the high tone, more prominent in CS at Cz and Pz, except for delta-ERD in PP at C3'. For both groups, we detected predominantly theta-ERD and alpha-ERD following both tone types within this second interval. PP showed more expressed theta-ERD at Fz and parietal theta-ERS. Alpha-ERD was significantly higher in CS, while frontal alpha-ERD was more prominent in the PP in response to both tones. The data obtained showed specific functional differences of event-related oscillatory activity in cognitive and sensory-motor information processing between the PP and CS. PMID- 19291393 TI - Memantine prevents sensitivity to excitotoxic cell death of rat cortical neurons expressing human truncated tau protein. AB - We have previously shown that human misfolded tau proteins strongly perturb mitochondrial transport and induce accumulation of free radicals in neurons. This interference is underlying cause of increased susceptibility to oxidative stress and could be linked to excitotoxic pathways. In order to understand integral mechanisms of misfolded tau driven neurodegeneration, we have investigated the role of human truncated tau protein, derived from Alzheimer's disease, in rat cortical neurons under the conditions of excitotoxic stress induced by glutamate. We found that primary neurons expressing truncated tau protein are highly susceptible to glutamate-induced cell death. Pre-treatment with memantine (N methyl-D-aspartate receptor antagonist) significantly improved survival of rat neurons exposed to glutamate and its effect was associated with overall decrease of reactive oxygen species (ROS) in both transgenic and nontransgenic neurons. Interestingly, despite of this overall effect, memantine was not able to decrease misfolded tau-induced ROS level specifically in transgenic cells. Our data suggest that memantine does not interfere with specific pathological pathways induced by misfolded tau protein nevertheless is able to attenuate oxidative stress in neurons. PMID- 19291394 TI - Role of radial glia in transformation of the primitive lumen to the central canal in the developing rat spinal cord. AB - In the last quarter of the embryonic development of rat and shortly after a termination of neurogenesis, the transformation of the spinal cord primitive lumen (pL) to the central canal (CC) occurs. In this work, we show that this phenomenon is not an insignificant event but it is directly associated with the processes of gliogenesis. Using a light microscopy and immunohistochemistry, we monitored the development of the rat embryonic spinal cord from the end of the neurogenesis on the embryonic day 17 until the maturation of the spinal cord during the first postnatal weeks. Our observations demonstrate the importance of the transformation of the pL to the CC and its connection with gliogenesis, and the mechanism of this transformation is proposed. It is found that a segregation of the glutamate transporter (GLAST) immunopositive cells from the alar plates and transformation of the radial glial cells to the fibrous and protoplasmic astrocytes play presumably a key role in the diminution of the ventricular zone. Results indicate that the very transformation and migration of the radial glial cells during gliogenesis could result in a transformation of the pL to the CC. PMID- 19291395 TI - Spinal cord transection significantly influences nNOS-IR in neuronal circuitry that underlies the tail-flick reflex activity. AB - AIM: Spinal cord transection interrupts supraspinal input and leads to the development of prominent spasticity. In this study, we investigated the effect of rat spinal cord transection performed at low thoracic level on changes in (i) neuronal nitric oxide synthase immunoreactivity (nNOS-IR), and (ii) the level of neuronal nitric oxide synthase (nNOS) protein in the neuronal circuitry that underlies tail-flick reflex. METHODS: nNOS-IR was detected by immunohistochemistry and the level of nNOS protein was determined by the Western blot analysis. The tail-flick reflex was tested by a noxious thermal stimulus delivered to the tail of experimental animals. After surgery, experimental animals survived for 7 days. RESULTS: A significant increase in the level of nNOS protein was found 1 week after thoracic transection in the L2-L6 segments. Immunohistochemical analysis discovered that this increase may be a result of (1) a high nNOS-IR in a large number of axons, located predominantly in the dorsal columns (DCs) of lower lumbosacral segments, and (2) a slight increase of density in nNOS-IR in motoneurons. On the other hand the number of nNOS-IR neurons in the superficial dorsal horn and in area surrounded the central canal (CC) was greatly reduced. The tail-flick response was immediate in animals after spinal transection, while control rats responded to thermal stimulus with a slight delay. However, the tail-flick latency in experimental animals was significantly higher than in control. CONCLUSION: These data indicate that transection of the spinal cord significantly influences nNOS-IR in neuronal circuitry that underlies the tail-flick reflex activity. PMID- 19291396 TI - Derivation of molecular signatures for breast cancer recurrence prediction using a two-way validation approach. AB - Previous studies have demonstrated the potential value of gene expression signatures in assessing the risk of post-surgical breast cancer recurrence, however, many of these predictive models have been derived using simple computational algorithms and validated internally or using one-way validation on a single dataset. We have recently developed a new feature selection algorithm that overcomes some limitations inherent to high-dimensional data analysis. In this study, we applied this algorithm to two publicly available gene expression datasets obtained from over 400 patients with breast cancer to investigate whether we could derive more accurate prognostic signatures and reveal common predictive factors across independent datasets. We compared the performance of three advanced computational algorithms using a robust two-way validation method, where one dataset was used for training and to establish a prediction model that was then blindly tested on the other dataset. The experiment was then repeated in the reverse direction. Analyses identified prognostic signatures that while comprised of only 10-13 genes, significantly outperformed previously reported signatures for breast cancer evaluation. The cross-validation approach revealed CEGP1 and PRAME as major candidates for breast cancer biomarker development. PMID- 19291397 TI - Involvement of matrix metalloproteinases on the inhibition of cells invasion and migration by emodin in human neuroblastoma SH-SY5Y cells. AB - Emodin (1,3,8-trihydroxy-6-methylanthaquinone), an active component present in the root and rhizome of Rheum palmatum L. (Polygonaceae) has anti-bacterial, anti tumor, diuretic and vasorelaxant effects. However, its mechanism of action on the cell migration and invasion of human neuroblastoma cancer SH-SY5Y cells is not fully understood. In this study, firstly, the effects of emodin on the percentage of viable cells were examined by using MTT assay and it was found that emodin induced dose-and time-dependent inhibition in human neuroblastoma SH-SY5Y cells. Second, the effects of emodin on the migration and invasion of SH-SY5Y cells were examined by using wound assay and matrigel counting and the results showed that emodin suppressed the migration and invasion of SH-SY5Y cells. Third, we examined the effect of emodin on the levels of associated proteins by using Western blotting and the results indicated that emodin inhibited the levels of GRB2, RhoA, HIF-1alpha, VEGF, FAK, iNOS, COX2, p-p38, p-c-jun, MMP2, MMP9 and MMP7 but promoted the levels of PKC, PI3K, MEKK3 and NF-kappaB p65 that led to the inhibition of migration and invasion of SH-SY5Y cells in vitro. PMID- 19291398 TI - Deciding versus implementing: a comment on "What gets noticed: how barrier and facilitator perceptions relate to the adoption and implementation of innovative mental health practices". PMID- 19291399 TI - Exploring the mental health of black men who have sex with men. AB - Current research indicates that black men who have sex with men (MSM) are disproportionately burdened by depressive distress and anxiety disorders as compared to their white gay and heterosexual counterparts. This study utilizes focus groups to qualitatively explore issues surrounding the mental health status of this population in an attempt to shed light on potential influencing and determinant factors. Twenty-two self-identified black, or multi-racial including black, MSM residing in Atlanta, Georgia participated in two focus groups--11 subjects each, respectively. Categories that emerged from data analysis include: knowledge/experiences, attitudes/beliefs, societal action/behavior, identity development, relationship functionality, and mental health status. Overarching themes for each category were delineated. PMID- 19291400 TI - Reproducibility and performance characteristics of colonic compliance, tone, and sensory tests in healthy humans. AB - BACKGROUND: The performance characteristics of colonic sensation and motility measurements are unclear. AIM: To compare left colon compliance, tone, and sensation in males and females and to evaluate inter and intra-individual coefficients of variation (COV) in these measurements. METHODS: Data were acquired using standard barostat methods, by one technologist, in 72 human volunteers (38 males, 18-65 years). We measured compliance, fasting tone, and sensation during baseline and post-placebo; postprandial (PP) tone was measured only post-placebo. Compliance and thresholds for first sensation, gas, and pain were measured using ascending method of limits; sensory ratings (0-100 mm VAS) using random phasic distensions at 8-36 mmHg above baseline operating pressure. Change in PP tone was measured by barostat balloon volume for the first 30 min after a 1000-kcal meal. Inter-COV was calculated as 100 (SD/mean), and intra-COV as (100 x SD delta/overall mean). RESULTS: There were no statistically significant associations with gender for most sensory or motor data at baseline. A modest association of fasting colonic tone and gender was observed. COV are lower (20-35%) for compliance, fasting tone, pain threshold, and sensation ratings than for PP tone and threshold for first or gas sensation (>45%). COV data are similar in males and females; sensation COVs appear smaller in females than in males. CONCLUSIONS: Testing of compliance, tone and pain, and gas sensation in left colon performs adequately to assess these functions in humans. Lower COV for sensation tests among females is relevant to plan studies of drugs intended for functional GI disorders. PMID- 19291401 TI - The link between post-traumatic stress disorder and physical comorbidities: a systematic review. AB - CONTEXT: Returning veterans from Afghanistan and Iraq will increase frequency of post-traumatic stress disorder (PTSD). Little is known about its impact on physical health. OBJECTIVE: Systematic literature review focusing on the association between PTSD and specific physical disorders. DATA SOURCES: An electronic search using PUBMED and hand search of four journals with an anxiety focus for studies published between January 1981 and July 2008, plus a manual search of article bibliographies. STUDY SELECTION: Original research reports focusing on PTSD and its association with physical health. Studies investigating only PTSD symptoms, trauma and physical disorders classified at the organ-system level were excluded. Eighty studies were reviewed and seven selected for final analysis. DATA EXTRACTION: Specific physical-health diagnoses were organized by system and tabulated. They were considered positive only if results were statistically significant. Total number of positive and negative studies for each diagnosis was then calculated for review. RESULTS: Seven studies examined the relationship between PTSD and specific physical disorders. Arthritis was associated with PTSD in most studies. Data conflicted regarding diabetes, coronary heart disease, and stroke. CONCLUSIONS: Few studies have examined the relationship between PTSD and physical health. Large, prospective epidemiological trials are needed. PMID- 19291402 TI - Modeling growth and quorum sensing in biofilms grown in microfluidic chambers. AB - Biofilms are highly organized structures coordinately formed by multiple species of bacteria. Quorum sensing (QS) is one cell-cell communication mechanism that is used by bacteria during biofilm formation. Biofilm formation is widely acknowledged to occur through a sequence of spatially and temporally regulated colonization events. While several mathematical models exist for describing biofilm development, these have been developed for open systems and are not applicable to closed systems where biofilm development and hydrodynamics are interlinked. Here, we report the development of a mathematical model describing QS and biofilm formation in a closed system such as a microfluidic channel. The model takes into account the effect of the external environment viz the mass and momentum transport in the microfluidic channel on QS and biofilm development. Model predictions of biofilm thickness were verified experimentally by developing Pseudomonas aeruginosa PA14 biofilms in microfluidic chambers and reflect the interplay between the dynamics of biofilm community development, mass transport, and hydrodynamics. Our QS model is expected to guide the design of experiments in closed systems to address spatio-temporal aspects of QS in biofilm development and can lead to novel approaches for controlling biofilm formation through disruption of QS spatio-temporal dynamics. PMID- 19291403 TI - A preliminary study of the population-adjusted effectiveness of substance abuse prevention programming: towards making IOM program types comparable. AB - The Institute of Medicine distinguishes between programs based on who is targeted: the entire population (universal), those at risk (selective), or persons exhibiting the early stages of use or related problem behavior (indicated). Evaluations suggest that although universal programs can be effective in reducing and preventing substance use, selective and indicated programs are both more effective and have greater cost-benefit ratios. This paper tests these assumptions by comparing the impact of these program types in reducing and preventing substance use at the individual level (i.e., those exposed to intervention services) and in the population (i.e., those exposed and not exposed to intervention services). A meta-analysis was performed on 43 studies of 25 programs to examine program comparability across IOM categories. When examining unadjusted effect sizes at the individual level, universal programs were modestly more successful in reducing tobacco use, but selective and indicated programs were modestly more successful in reducing alcohol and marijuana use. When adjusted to the population level, the average effect sizes for selective and indicated programs were reduced by approximately half. At the population level, universal programs were more successful in reducing tobacco and marijuana use and selective and indicated programs were more successful in reducing alcohol use. Editors' Strategic Implications: the authors' focus on the public health value of a prevention strategy is compelling and provides a model for analyses of other strategies and content areas. PMID- 19291404 TI - Competition effects in phonological priming: the role of mismatch position between primes and targets. AB - In three experiments, we examined lexical competition effects using the phonological priming paradigm in a shadowing task. Experiments 1A and 1B showed that an inhibitory priming effect occurred when the primes mismatched the targets on the last phoneme (/bagar/-/bagaj/). In contrast, a facilitatory priming effect was observed when the primes mismatched the targets on the medial phoneme (/viraj/-/vilaj/). Experiment 2 replicated these findings with primes presented visually rather than auditorily. The data thus indicate that the position of the mismatching phoneme is a critical factor in determining the competition effect between prime and target words. PMID- 19291405 TI - Religion, evolution, and mental health: attachment theory and ETAS theory. AB - This article reviews the historical origins of Attachment Theory and Evolutionary Threat Assessment Systems Theory (ETAS Theory), their evolutionary basis and their application in research on religion and mental health. Attachment Theory has been most commonly applied to religion and mental health in research on God as an attachment figure, which has shown that secure attachment to God is positively associated with psychological well-being. Its broader application to religion and mental health is comprehensively discussed by Kirkpatrick (2005). ETAS Theory explains why certain religious beliefs--including beliefs about God and life-after-death--should have an adverse association, an advantageous association, or no association at all with mental health. Moreover, it makes specific predictions to this effect, which have been confirmed, in part. The authors advocate the application of ETAS Theory in research on religion and mental health because it explains how religious and other beliefs related to the dangerousness of the world can directly affect psychiatric symptoms through their affects on specific brain structures. PMID- 19291406 TI - "And let us make us a name": reflections on the future of the religion and health field. AB - After years of marginality, research on religion and health is entering the academic mainstream. Scholarship on this topic has evolved into a large, productive field. As in any emerging field, there are competing visions for what the field should be about and what research questions should be pursued. Different opinions exist as to which constructs should be researched. Words like religion, spirituality, faith, and prayer, and health, healing, medicine, and healthcare, imply different things. The study of their various interconnections can thus take myriad forms. This article argues for a welcoming approach open to the widest range of research subjects. PMID- 19291407 TI - Clearance of apoptotic corpses. AB - Apoptotic corpses can be engulfed and cleared by many other cell types in addition to 'professional' phagocytes such as macrophage. Studies of several organisms have contributed to the understanding of apoptotic corpse engulfment. Two partially redundant engulfment pathways have been characterized that act even in non-professional phagocytes to promote corpse engulfment. This review summarizes some recent progress in signaling by these pathways, including the exposure of eat-me-signals on apoptotic cells, and insights from Drosophila on the roles of the bridging receptor Six Microns Under, the non-receptor tyrosine kinase Shark, and store-operated calcium release in the Draper/Ced-1 pathway of corpse recognition and internalization. The mechanism of apoptotic phagosome maturation is outlined, and possible connections between corpse engulfment and proliferation, cell competition, and immunity are discussed. PMID- 19291408 TI - High efficient electrical stimulation of hippocampal slices with vertically aligned carbon nanofiber microbrush array. AB - Long-term neuroprostheses for functional electrical stimulation must efficiently stimulate tissue without electrolyzing water and raising the extracellular pH to toxic levels. Comparison of the stimulation efficiency of tungsten wire electrodes (W wires), platinum microelectrode arrays (PtMEA), as-grown vertically aligned carbon nanofiber microbrush arrays (VACNF MBAs), and polypyrrole coated (PPy-coated) VACNF MBAs in eliciting field potentials in the hippocampus slice indicates that, at low stimulating voltages that preclude the electrolysis of water, only the PPy-coated VACNF MBA is able to stimulate the CA3 to CA1 pathway. Unlike the W wires, PtMEA, as-grown VACNF MBA, and the PPy-coated VACNF MBA elicit only excitatory postsynaptic potentials (EPSPs). Furthermore, the PPy coated VACNF MBA evokes somatic action potentials in addition to EPSPs. These results highlight the PPy-coated VACNF's advantages in lower electrode impedance, ability to stimulate tissue through a biocompatible chloride flux, and stable vertical alignment in liquid that enables access to spatially confined regions of neuronal cells. PMID- 19291409 TI - Fabrication and physical evaluation of a polymer-encapsulated paramagnetic probe for biomedical oximetry. AB - Lithium octa-n-butoxynaphthalocyanine (LiNc-BuO) is a promising probe for biological electron paramagnetic resonance (EPR) oximetry and is being developed for clinical use. However, clinical applicability of LiNc-BuO may be hindered by potential limitations associated with biocompatibility, biodegradation, and migration of individual crystals in tissue. To overcome these limitations, we have encapsulated LiNc-BuO crystals in polydimethyl siloxane (PDMS), an oxygen permeable and bioinert polymer, to fabricate conveniently implantable and retrievable oxygen-sensing chips. Encapsulation was performed by a simple cast molding process, giving appreciable control over size, shape, thickness and spin density of chips. The in vitro oxygen response of the chip was linear, reproducible, and not significantly different from that of unencapsulated crystals. Cast-molding of the structurally-flexible PDMS enabled the fabrication of chips with tailored spin densities, and ensured non-exposure of embedded LiNc BuO, mitigating potential biocompatibility/toxicological concerns. Our results establish PDMS-encapsulated LiNc-BuO as a promising candidate for further biological evaluation and potential clinical application. PMID- 19291410 TI - School personnel perspectives on their school's implementation of a school-based suicide prevention program. AB - Youth suicide is a national public health priority, with policymakers highlighting schools as an ideal setting in which to deliver suicide prevention programs. Over the past decade, the number of schools implementing such programs has grown substantially, yet little is known about how successfully such programs are being implemented. This study examines the implementation of a district-wide suicide prevention program through key informant interviews with school personnel. Schools with higher rates of implementing district protocols for at risk students had an organized system to respond to at-risk students, a process for effectively responding to students who were at-risk for suicide, and strong administrative support. In contrast, schools that had lower rates of implementing district protocols relied on a handful of individuals for suicide prevention activities and had limited administrative support. Attention to organizational factors leading to successful implementation of school-based suicide prevention programs may enhance the role of schools in national adolescent suicide prevention efforts. PMID- 19291411 TI - The abbreviated impactor measurement (AIM) concept: part II--Influence of evaporation of a volatile component-evaluation with a "droplet-producing" pressurized metered dose inhaler (pMDI)-based formulation containing ethanol as cosolvent. AB - The abbreviated impactor measurement (AIM) concept is a potential solution to the labor-intensive full-resolution cascade impactor (CI) methodology for inhaler aerosol aerodynamic particle size measurement. In this validation study, the effect of increasing the internal dead volume on determined mass fractions relating to aerodynamic particle size was explored with two abbreviated impactors both based on the Andersen nonviable cascade impactor (ACI) operating principle (Copley fast screening Andersen impactor [C-FSA] and Trudell fast screening Andersen impactor [T-FSA]). A pressurized metered dose inhaler-delivered aerosol producing liquid ethanol droplets after propellant evaporation was chosen to characterize these systems. Measures of extrafine, fine, and coarse particle mass fractions from the abbreviated systems were compared with corresponding data obtained by a full-resolution ACI. The use of liquid ethanol-sensitive filter paper provided insight by rendering locations visible where partly evaporated droplets were still present when the "droplet-producing" aerosol was sampled. Extrafine particle fractions based on impactor-sized mass were near equivalent in the range 48.6% to 54%, comparing either abbreviated system with the benchmark ACI-measured data. The fine particle fraction of the impactor-sized mass determined by the T-FSA (94.4 +/- 1.7%) was greater than using the C-FSA (90.5 +/ 1.4%) and almost identical with the ACI-measured value (95.3 +/- 0.4%). The improved agreement between T-FSA and ACI is likely the result of increasing the dead space between the entry to the induction port and the uppermost impaction stage, compared with that for the C-FSA. This dead space is needed to provide comparable conditions for ethanol evaporation in the uppermost parts of these impactors. PMID- 19291412 TI - Recent advances in structure-based virtual screening of G-protein coupled receptors. AB - In addition to the rhodopsin crystal structure, high-resolution crystal structures of ligand-mediated G-protein-coupled receptors (GPCRs) have recently become available, and these have become attractive templates for developing homology models of several GPCRs of therapeutic interest. These crystal structures and the homology models derived from them have provided significant insights into ligand-receptor interactions. Moreover, several studies have demonstrated that the structural models are indeed suitable for virtual screening of compound databases to identify new ligands for various GPCRs. Recent examples of such virtual screening against GPCRs are discussed in this review. PMID- 19291413 TI - Validation of the Chinese version of Underwood's Daily Spiritual Experience Scale -transcending cultural boundaries? AB - BACKGROUND: Daily spiritual experience (DSE) refers to one's interaction with the transcendent in day-to-day life. Underwood's Daily Spiritual Experience Scale mic(DSES) was developed to measure this experiential component of religiousness and spirituality. Addressing ordinary daily experiences rather than particular beliefs, DSES has transcultural applicability potential. PURPOSE: The current study aimed to develop and evaluate the Chinese version of DSES (DSES-C). METHOD: The 16-item scale was translated faithfully through standard translation/back translation procedures. The term "God" required an extended definition embracing both a humanized and a philosophical higher power in Chinese culture. The translated scale plus a battery of validation scales were administered to staff of a large rehabilitation service complex, resulting in 245 completed questionnaires. RESULTS: Exploratory factor analysis revealed a similar factor structure as the original English version and similar problems with items 13 (compassion) and 14 (mercy). After carefully deliberating on the Chinese conceptualization of spirituality and balancing psychometric properties, the one factor 16-item structure of the English version was supported. DSES-C showed high internal consistency (Cronbach's alpha = 0.97). Construct validity was supported by correlations with validation scales in expected directions. CONCLUSION: The psychometric properties of DSES were similar to the English version in factor structure, internal consistency, and convergence/divergence construct validity. PMID- 19291414 TI - Soil factors associated with zinc deficiency in crops and humans. AB - Zinc deficiency is the most ubiquitous micronutrient deficiency problem in world crops. Zinc is essential for both plants and animals because it is a structural constituent and regulatory co-factor in enzymes and proteins involved in many biochemical pathways. Millions of hectares of cropland are affected by Zn deficiency and approximately one-third of the human population suffers from an inadequate intake of Zn. The main soil factors affecting the availability of Zn to plants are low total Zn contents, high pH, high calcite and organic matter contents and high concentrations of Na, Ca, Mg, bicarbonate and phosphate in the soil solution or in labile forms. Maize is the most susceptible cereal crop, but wheat grown on calcareous soils and lowland rice on flooded soils are also highly prone to Zn deficiency. Zinc fertilizers are used in the prevention of Zn deficiency and in the biofortification of cereal grains. PMID- 19291415 TI - Medical demography and epidemiology: dizygotic twins. AB - Demography is the statistical study of populations, epidemiology of health of populations. Fascinating challenges for future research, shared by epidemiology and demography, are to be found in the human life course, how the life course is transmitted over generations, how the healthy life is shaped by dynamics within the life course and how the life course is affected by the social and economic environment. The life course begins when two future parents meet. Then social disadvantage or privilege is beginning and diverse rates of ageing are being fixed. Health in utero and at young age will co-determine health at middle and old age, among others mediated by a greater intelligence and a better education. Particularly better education compresses cognitive disability at the end of life, building cognitive reserves that increase brain plasticity and enhance resistance to damage. While with extended life spans may come extended life spans with disability, the twin demography and epidemiology may show how to lighten the burden of senescence at the end of life. PMID- 19291416 TI - Colloid in the anterior pituitary of helmet guinea fowl (Numida meleagris galeata): morphometric analysis and pattern of occurrence in relation to apoptosis. AB - Colloidal accumulations in the pars distalis of helmet guinea fowls at various ages from 1 to 450 days were examined by Periodic acid-Schiff reaction, immunohistochemistry and electron microscopy. Round, ovoid and elongated colloids were observed. Colloids (69.5 +/- 2.997) with 0.169 +/- 0.014 microm mean diameter were already present in a 1-day-old bird. Numerous colloids were encountered in 450 days old birds (2931.333 +/- 29.847) with 2.263 +/- 0.078 microm mean diameter of round colloids. A significant difference in the mean colloidal number and diameter between young and adult birds was observed. In young birds (aged 1-30 days) both Periodic acid-Schiff reaction positive colloids and S-100 positive folliculostellate (FS) cells were found to appear first on or near the posterolateral region. In adult birds, FS cells were found to completely surround the colloids. We examined the biochemical components of colloids and the relationship with apoptosis by immunohistochemistry. Results showed that the colloids are composed of clusterin protein. Apoptotic cells detected by single stranded DNA (ssDNA) were abundant and localized preferentially near colloids. To define clearly the type of cells undergoing apoptosis in the anterior pituitary, we performed electron microscopy. Numerous endocrine cells at different stages of apoptosis were found engulfed by FS cells that were in close association with the colloidal accumulations. The occurrence of extremely large number of colloids in relation to apoptotic profiles in anterior pituitary of helmet guinea fowl is discussed. PMID- 19291417 TI - A investigation of border disease virus in sheep in Western Turkey. PMID- 19291418 TI - Analytical approaches to photobiological hydrogen production in unicellular green algae. AB - Several species of unicellular green algae, such as the model green microalga Chlamydomonas reinhardtii, can operate under either aerobic photosynthesis or anaerobic metabolism conditions. A particularly interesting metabolic condition is that of "anaerobic oxygenic photosynthesis", whereby photosynthetically generated oxygen is consumed by the cell's own respiration, causing anaerobiosis in the culture in the light, and induction of the cellular "hydrogen metabolism" process. The latter entails an alternative photosynthetic electron transport pathway, through the oxygen-sensitive FeFe-hydrogenase, leading to the light dependent generation of molecular hydrogen in the chloroplast. The FeFe hydrogenase is coupled to the reducing site of photosystem-I via ferredoxin and is employed as an electron-pressure valve, through which electrons are dissipated, thus permitting a sustained electron transport in the thylakoid membrane of photosynthesis. This hydrogen gas generating process in the cells offers testimony to the unique photosynthetic metabolism that can be found in many species of green microalgae. Moreover, it has attracted interest by the biotechnology and bioenergy sectors, as it promises utilization of green microalgae and the process of photosynthesis in renewable energy production. This article provides an overview of the principles of photobiological hydrogen production in microalgae and addresses in detail the process of induction and analysis of the hydrogen metabolism in the cells. Furthermore, methods are discussed by which the interaction of photosynthesis, respiration, cellular metabolism, and H(2) production in Chlamydomonas can be monitored and regulated. PMID- 19291419 TI - Electroweak parity-violating energy shifts of amino acids: the "conformation problem". AB - The preceding paper described our coupled-perturbed Hartree-Fock (CPHF) and density functional theory (DFT) methods of computing the parity-violating energy shift (PVES). This paper addresses the "conformation problem"-the difficulty determining which hand of amino acids in solution is favoured by the weak force due to the difficulty determining the solution conformation. We attempt to resolve this by using the methods of the preceding paper to compute the PVES of solution and gas-phase amino acid structures determined by other groups from high level optimizations that include solvation. We conclude that the conformational hypersensitivity of the PVES still precludes a definite conclusion as to the sign of the PVES of L-alanine in solution, but that there is no problem in the gas phase: the PVES of gas-phase L-alanine is decisively negative. We show that the PVES is very sensitive to certain torsion angles, but is not hypersensitive to bondlengths or bond angles. In determining structures for PVES computations, there is therefore no need for expensive full optimizations: one can just optimize the crucial torsion angles. We present new computations of gas-phase amino acids PVESs, using partial optimizations with small basis sets, and the results agree well with those from higher level techniques. In the following paper we apply these less costly techniques to larger amino acids. The "conformation problem" has led some to dismiss the PVES as the source of life's handedness, but we believe this is premature: we show here that amino acids are a special case because their favoured conformations are almost achiral. PMID- 19291420 TI - Inborn errors of purine and pyrimidine metabolism. AB - Genetic disorders of purine and pyrimidine (PP) metabolism are under-reported and infrequently mentioned in the general literature, as well as in reviews dedicated to other inborn errors of metabolism. Owing to limited awareness, relatively recent recognition, as well as considerable phenotypic variation, these disorders may often be misdiagnosed or remain undiagnosed. Disorders that arise as a result of dysfunction in PP metabolism represent some of the most challenging diagnostic problems in medicine. In addition to their low prevalence rates, they also present with extremely variable signs and symptoms. They may affect any system in a variety of manners, and often mimic other, more recognizable disorders. The diagnostic problem is compounded by the fact that some biochemically affected patients are symptom-free. Rapidly evolving laboratory techniques such as high performance liquid chromatography coupled to tandem mass spectrometry are now well established as the preferred method for detection for these defects, but currently the most important step in diagnosis consists of suspecting the disorder. Diagnosis is vital because genetic counselling can be provided and, in some cases, specific treatment can be offered that may slow or even reverse clinical symptoms. If undiagnosed, these disorders can be devastating to patients and their families, resulting in early death or institutionalization for the rest of patient's life. This article describes the current state of knowledge about inborn errors of purine and pyrimidine metabolism, focusing on the varying clinical presentations, the laboratory findings and discusses indications for selective screening for these disorders. PMID- 19291421 TI - Magnetic resonance imaging of persistent myocardial obstruction after myocardial infarction. A tool becoming increasingly important in clinical cardiology? PMID- 19291422 TI - Signal intensity enhances diagnostic capacity in myocardial infarction. PMID- 19291423 TI - Heat stress upregulates chaperone heat shock protein 70 and antioxidant manganese superoxide dismutase through reactive oxygen species (ROS), p38MAPK, and Akt. AB - Chinese hamster lung fibroblasts V79 cells were treated with heat stress for 4 weeks with short duration (15 min) heat shock every alternate day in culture. It was observed that Hsp 70 and the antioxidant enzyme MnSOD became overexpressed during the chronic heat stress period. Both p38 MAPK and Akt became phosphorylated by chronic heat stress exposure. Simultaneous exposure to SB203580, a potent and specific p38MAPK inhibitor drastically inhibited the phosphorylation of p38MAPK and Akt. Furthermore, exposure to SB203580 also blocked the increase in Hsp70 and MnSOD levels and the elevated SOD activity brought about by chronic heat stress. Heat shock factor 1 (HSF1) transcriptional activity and nuclear translocation of HSF1 were prominently augmented by chronic heat stress, and this amplification is markedly reduced by concomitant exposure to SB203580. Also, activations of p38MAPK and Akt and upregulations of Hsp70 and MnSOD were observed on exposure to heat shock for a single exposure of longer duration (40 min). siRNA against p38MAPK notably reduced Akt phosphorylation by single exposure to heat stress and drastically diminished the rise in Hsp70 and MnSOD levels. Similarly, siRNA against Akt also eliminated the augmentation in Hsp70 and MnSOD levels but p38MAPK levels remained unaffected. Heat stress produced reactive oxygen species (ROS) in V79 fibroblasts. N-acetyl cysteine blocked the increase in phosphorylation of p38MAPK, amplification of Hsp70, and MnSOD levels by heat stress. Therefore, we conclude that heat stress-activated p38MAPK which in turn activated Akt. Akt acted downstream of p38MAPK to increase Hsp70 and MnSOD levels.Concise summary: Thermal injury of the skin over a long period of time has been associated with development of cancerous lesions. Also, in many cancers, the cytoprotective genes Hsp70 and MnSOD have been found to be overexpressed. Therefore, we considered it important to identify the signaling elements upstream of the upregulated survival genes in heat stress. We conclude that heat stress activated p38MAPK which in turn activated Akt. Akt mediated an augmentation in Hsp70 and MnSOD levels working downstream of p38MAPK. PMID- 19291424 TI - Regulatory T cells in type 1 diabetic patients with autoimmune chronic atrophic gastritis. AB - Type A chronic atrophic gastritis (CAG) is increased in type 1 diabetic patients (DM1). To address this issue, we determined and analyzed the number of peripheral blood regulatory T cells (Tregs) in 15 DM1-CAG patients, 15 DM1 patients without associated autoantibodies (DM1) and 15 healthy controls by flow cytometry and compared gastric Tregs expression (CD4+Foxp3+/CD4+) in DM1-CAG patients with that observed in 10 control Helicobacter pylori CAG-infected biopsies. The percentage of peripheral Tregs was higher in DM1-CAG patients compared to DM1 and controls (CD4+Foxp3+: 7.67 +/- 1.91% vs. 5.38 +/- 1.57% and 5.65 +/- 1.76%, P < 0.001, respectively), with no differences between DM1 and controls. Gastric mucosal Tregs were higher in H. pylori CAG than in DM1-CAG patients (31.31 +/- 5.52% vs. 7.68 +/- 3.70%; P < 0.001). Data suggest that Tregs are stimulated in patients with more than one autoimmune disease (DM1 + CAG) in an ineffectual attempt to control autoimmune response and that the number of Tregs in gastric mucosa implicated in the chronification of gastritis differs according to the etiology. PMID- 19291425 TI - Association of TRB3 gene Q84R polymorphism with type 2 diabetes mellitus in Chinese population. AB - BACKGROUND: TRB3, a human homolog of Drosophila Tribbles, has been shown as a critical negative regulator of Akt (also known as protein kinase B), which is a key component in insulin signaling. In addition, TRB3 is another PPAR-target gene and functions as an important link between glucose and lipid metabolism. The Q84R polymorphic variant of TRB3 has been linked to insulin resistance and related clinical outcomes. However, it is unclear whether this polymorphism is associated with type 2 diabetes mellitus (T2DM) in the Chinese population. METHODS: In this study, we genotyped Q84R polymorphism in 177 patients with T2DM and 245 control subjects in Chinese population by using the polymerase chain reaction/ligase detection reaction (PCR/LDR) assay. RESULTS: No significant difference in the Q84R genotype frequency was observed between T2DM patients and controls (P = 0.642). In T2DM group, the Q84R variant in cases was associated with higher FINS, higher HOMA-IR, and lower LnISI (P = 0.003, 0.001, and 0.001, respectively). However, the changes in HOMA-IR and LnISI were not significant in controls (the P value is 0.098 and 0.203, respectively). In addition, FINS levels were also significantly increased from Q84Q to R84 in controls (P = 0.036). CONCLUSION: Our data indicate that the TRB3 Q84R polymorphism is not associated with T2DM in Chinese population. However, the Q84R variant is associated with insulin resistance among T2DM patients in Chinese population. PMID- 19291426 TI - Suberoylanilide hydroxamic acid (SAHA) induces growth arrest and apoptosis in pituitary adenoma cells. AB - Pituitary adenomas, which account for 15-20% of intracranial tumors, are associated with significant morbidity and mortality, due in part, to hormone hypersecretion and mass effects following increased proliferation. Radiotherapy and surgery remain frontline treatment options; however, adverse side effects and surgical limitations to treat invasive tumors necessitate the need for novel therapeutic targets. This study tested the efficacy of the histone deacetylase inhibitor (HDACi), suberoylanilide hydroxamic acid (SAHA) in GH3, and MMQ pituitary adenoma cells. Clinically achievable concentrations of SAHA (500 nM-4 microM) induced growth arrest and increased cell death in GH3 pituitary adenoma cells. Further investigation into the mechanism of cell death revealed an increase in PARP cleavage and procaspase-3 activation, consistent with apoptotic cell death. SAHA also attenuated the expression of anti-apoptotic IAP (XIAP, survivin) and Bcl-2 family proteins (Bcl-2, Bcl-xL), but did not alter Bax expression. Together, these findings support a possible utility for SAHA alone or in combination with radiation for the treatment of pituitary adenoma. PMID- 19291427 TI - Development of a RT real-time PCR for the detection and quantification of human rhinoviruses. AB - Human Rhinoviruses (HRV) are the most common viral agents, being responsible for upper as well as lower respiratory tract infections. Evidence demonstrating that HRV disease is not exclusively limited to the upper airways and may cause lower respiratory complications, together with the frequency of HRV infections and the increasing number of immunocompromised patients underline the need for including HRV in virological diagnostics of acute lower respiratory tract illness. This article describes the development and optimization of a reverse transcription (RT) real-time PCR assay for quantification of HRV RNA in clinical samples. Efficiency, sensitivity, specificity, inter- and intra-assay variability, and dynamic range have been determined. Subsequently, the assay has been validated on bronchoalveolar lavage (BAL) specimens obtained from immunocompetent and immunocompromised patients. PMID- 19291428 TI - Cancer detection and treatment: the role of nanomedicines. AB - Nanotechnology is a field which has been at the forefront of research over the past two decades. The full potential of nanotechnology has yet to be fully realized. One subset of nanotechnology that has emerged is nanomedicine, which has been able to exploit the unique properties of nano-sized particles for therapeutics. Nanomedicine has the potential to increase the specific treatment of cancer cells while leaving healthy cells intact through the use of novel nanoparticles to seek and treat cancer in the human body. However, there are undoubtedly toxicities, which have not yet been fully elucidated. Various nano carriers such as nanoshells, nanocrystals, nanopolymers, quantum dots, and dendrimers, and their role in early cancer detection and treatment have been discussed in this article. PMID- 19291429 TI - Immunohistochemical detection of the Raf kinase inhibitor protein in nonneoplastic gastric tissue and gastric cancer tissue. AB - Expression of the Raf kinase inhibitor protein (RKIP), a metastasis suppressor, is high in normal tissues, low in primary cancers, and lowest or absent in metastatic cancers. Here, we studied the expression of RKIP in nonneoplastic gastric tissue and gastric cancer tissue by immunohistochemistry (IHC) and evaluated its role in the genesis and metastasis of gastric cancer. RKIP immunoreactivity was evaluated in 40 samples of nonneoplastic gastric tissues and 75 samples of gastric cancer tissues. Among the 40 samples of nonneoplastic gastric tissue, 35 (87.5%) were positive for RKIP expression and 5 (12.5%) were negative; in the 75 samples of primary gastric cancer tissue, 39 (52%) were positive for RKIP expression and 36 (48%) were negative. Among 26 samples of metastatic lymph node tissues, 5 (19.2%) were positive for RKIP expression and 21 (80.8%) were negative. RKIP expression level was highest in nonneoplastic gastric tissue, low in primary gastric cancer tissue, and lowest or undetectable in metastatic gastric cancer tissue. Our data suggest that RKIP may play a role in the genesis and metastasis of gastric cancer. PMID- 19291430 TI - Determination of cyanide in blood by reaction head-space gas chromatography. AB - A method describing determination of cyanide in blood by head-space gas chromatography with electron capture detector was reported. The method involves transformation of cyanide into cyanogen chloride by reacting hydrogen cyanide with chloramine-T on a stick of filter paper in the space above the blood in the head-space vial. The recovery was 84-96% and the coefficient of variation was 3.3 7.2%. The limit of quantitation was about 0.01 mg cyanide/l. PMID- 19291431 TI - Intersecting fractures of the skull and gunshot wounds. Case report and literature review. AB - When two fracture lines of a solid surface (ice, glass, eggshell, etc.) intersect, it is always possible to tell which one has been made first. Indeed pre-existing damage of the surface arrests all the fracture lines produced by subsequent impacts. This well-known principle (established by Puppe in 1903) has been largely used in glass fracture analysis, but can be applied also to the examination of skull fractures. It can help sequencing blunt force or gunshot injuries determining the direction of fire and differentiating entrance from exit wounds in the absence of specific distinguishing features (i.e., internal/external beveling of the skull or overlying skin indicators). In this context, we report the case of a 76-year-old man who shot himself in the mouth with a Walther PPK 7.65 handgun and highlight the utility of the application of both Puppe's Rule and Multislice Computed Tomography (MSCT) in the examination of gunshot wounds to the skull. PMID- 19291432 TI - Sudden death by laryngeal polyp: a case report and review of the literature. AB - Sudden unexpected death in the context of bizarre or unusual behavior usually relates to acute drug intoxication or excited delirium. We report the case of a man who died suddenly while running naked on a public street. Although the initial death investigation was indicative of excited delirium or drug intoxication, autopsy revealed glottic obstruction by an inflammatory laryngeal polyp. Toxicologic studies were negative and investigation revealed presentation at a hospital the day before death with stridor. It is believed he developed airway obstruction while dressing at home and ran out on the street to find assistance. This case illustrates the importance of a thorough death investigation and a broad differential diagnosis when approaching a forensic autopsy. PMID- 19291433 TI - Electrical injury of the neck and cardiac air embolism: a novel mechanism of death. AB - We report a case of air embolism in an industrial worker complicating a deep electrical injury in the neck. He was found dead in an upright position while his chest and head were resting on top of a live transformer. At autopsy, there were multiple electrical burns involving the face, neck, chest, and arms. There was a deep electrical burn associated with hemorrhage in the neck involving the skin, underlying subcutaneous tissue, strap muscles, and the thyroid gland. The internal examination showed a prominently bulging right atrium, which was filled with air and not blood. At autopsy, when the pericardium was filled with water and the right atrium opened, a large number of bubbles emerged. We concluded that death was due to air embolism complicating electrical damage to the neck veins. This case illustrates that acute complications of electrical burns rather than electrocution-related cardiac arrhythmia can be the mechanism of death in rare electricity-related deaths. It also reveals the difficulties encountered by the pathologist in determining the exact mechanism of death when there are competing mechanisms. PMID- 19291436 TI - Homicidal smothering: vital histological confirmation of orofacial injury despite a prolonged post-mortem interval. AB - Homicidal smothering is the deliberate occlusion of the external airways, usually using the hands, pillows, or bedding. Victims are often those who are unable to resist, such as the young or the old. The limited resistance offered by these individuals may result in a comparative lack of injury. We present a case of a 72 year-old female, found deceased on her bed. The case was initially not considered suspicious and a coronial autopsy was performed. Concerns were subsequently raised and the body remained refrigerated. Six months after the initial examination, an opinion was requested as to whether further examination would be worthwhile to consider allegations of deliberate smothering. Review of the scene photographs showed deviation of the nose, eversion of the upper lip, and suspected intra-oral bruising. A bloodstained pillow was adjacent to the face. At the later autopsy, the body was found to show significant post-mortem deterioration and the facial tissues were mummified. The upper lip was excised and routinely processed for histology, sections showing remarkably good tissue preservation and extensive recent bruising, allowing the suspicious lip injuries in the scene photographs to be safely interpreted. This report illustrates the subtle pathological findings apparent in many of these cases and reinforces the need for thorough external examination and correlation with forensic scene investigation. Histological sampling of suspected injuries can be rewarding, even in the presence of severe post-mortem deterioration. PMID- 19291439 TI - From the Editor-in-Chief. PMID- 19291440 TI - Driving under the influence of gamma-hydroxybutyrate (GHB). AB - We used an in-house forensic toxicology database (TOXBASE) to evaluate the occurrences of gamma-hydroxybutyrate (GHB) in blood samples from people arrested in Sweden for driving under the influence of drugs (DUID) between 1998 and 2007. Age, gender, and concentrations of GHB in blood were compared and contrasted when GHB was the only drug present and when it occurred along with other drugs. GHB was determined in blood by gas chromatography (GC) after conversion to gamma butyrolactone (GBL) and analysis of the latter with a flame ionization detector. The cut-off concentration of GHB in blood for reporting a positive result was 8 mg/l, which served as limit of quantitation. The mean and median GHB concentrations were 89 mg/l and 82 mg/l, respectively (2(1/2) and 97(1/2) percentiles 12 and 220 mg/l) in 548 arrested drivers. These individuals were predominantly men (95%) with an average age of 26 +/- 5.5 years (range 15-50 years) and women (5%) were several years older with an average age of 32 +/- 8.0 years (range 19-47). There were 102 individuals (29%) who were arrested more than once with GHB in blood (average approximately 3 times per person) and one as many as 10 times. GHB was the only psychoactive substance detected in 215 cases (39%) at mean and median blood-concentrations of 91 mg/l and 83 mg/l, respectively. These concentrations were not significantly different from poly-drug users. A weak but statistically significant correlation existed between the concentration of GHB in blood and the person's age (N = 548, r = 0.135, P < 0.01). The signs of drug influence noted by arresting police officers included sedation, agitation, unsteady gait, slurred speech, irrational behavior, jerky body movements, dilated pupils, and spitting. The blood concentrations reported here are probably appreciably less than at time of driving (30-90 min earlier) owing to the short elimination half-life of GHB (t ((1/2)) = 30-40 min). PMID- 19291441 TI - Experimental simulation of non-ballistic wounding by sharp and blunt punches. AB - Despite a long history of gross and microscopic descriptions of blunt and sharp force injury to the dermal tissues, few have addressed the mechanisms underlying such trauma. The need to develop an understanding of how non-ballistic injury occurs calls for an ability to biomechanically model the process. We recently introduced a basic skin and subcutaneous model, which we used to investigate wounding from a spherical object. Here we employ the same model to examine wounding caused by a sharp wedge shaped object and a blunt rectangular object. Macroscopic examination and SEM views of the surface and cross sections of blunt and sharp force tears show that while in the former there is a clean cut through the skin into the underlying sponge, in the latter there is a tissue plug confined to the skin that is smaller than the impacting rectangle. Fracture initiation in the subdermal tissue occurs at the angles of the impacting object. In sharp force trauma, there is localized breaching of the skin layer coupled with the wedging action of the impacting object. Because the subdermal tissue, in this case the underlying hydrated foam, is attached to the base of the skin, it will contribute to further tearing of the foam beneath the line of contact. PMID- 19291442 TI - Suicidal and homicidal sharp force injury: a 5-year retrospective comparative study of hesitation marks and defense wounds. AB - In sharp force injury cases, the presence of hesitation marks or defense wounds figures among useful characteristics in the distinction of suicide and homicide. However, there are very few systematic comparative studies of hesitation marks and defense wounds in the forensic literature. This study was thus undertaken to systematically compare features of hesitation marks and defense wounds, with a particular emphasis on dispersion patterns. Over a 5-year period, all suicidal and homicidal sharp force cases were retrospectively reviewed for hesitation marks and defense wounds. A total of 58 suicides (7 women; 51 men) and 149 homicides (59 women; 60 men) were found, of which 74% (n = 43) were positive for hesitation marks and 61% (n = 91) for defense wounds. On the upper limbs, hesitations marks were more often observed on the anterior aspect of the limb, while defense wounds were equally distributed on the anterior and posterior aspect. For hand lesions, hesitation marks were generally located on one side only, while defense wounds more commonly involved both sides. No left or right predominance was observed in hesitations marks or defense wounds. Defense wounds were more widely distributed on the upper limbs than hesitation marks. PMID- 19291443 TI - Microscopical examination of particles on smoked cigarette filters. AB - Cigarette butts collected from crime scenes can play an important role in forensic investigations by providing a DNA link to a victim or suspect. Microscopic particles can frequently be seen on smoked cigarette filters with stereomicroscopy. The authors are not aware of previous published attempts to identify this material. These particles were examined with transmission and scanning electron microscopy and were found to consist of two types of superficial epithelial tissue, consistent with two areas of the lip surface. The particles were often composed of several layers of non-nucleated and nucleated epithelium with the former being the most common. It was further determined that both of these cell types are easily transferred from the lip. The results of this study indicate that the most visible source of DNA obtained from cigarette butts and other objects in contact with the lip may be lip epithelial tissue. PMID- 19291444 TI - Intrathoracic petechiae in SIDS: a retrospective population-based 15-year study. AB - Intrathoracic petechiae (IP), the most common gross finding in sudden infant death syndrome (SIDS) cases at autopsy, suggest upper airway obstruction (UAO) occurs during the terminal event. If true, IP would be expected more frequently among SIDS cases found face down compared to other face positions. We compare the rates of IP in SIDS cases found face down versus other face positions. A retrospective 15-year review of IP among all cases of SIDS occurring from 1991 through 2005 accessioned by the San Diego County Medical Examiner's Office was conducted. The presence or absence of IP was based on both gross and microscopic observations of the thoracic organs. The severity of thymic petechiae was scored semiquantitatively from microscopic observations. There were 489 SIDS cases during the study period. Sixteen of these were excluded because their deaths had been delayed by initially successful cardiopulmonary resuscitation. Among the remaining 473 SIDS cases, face position when found was specifically described for 332 (70%). Of 122 cases found face down, 112 (92%) had IP, compared to 85% (179) of 210 infants found with the face up or to the side (P = 0.06). The groups were not different with respect to age or gender, but the racial distribution was significantly different (P = 0.004). African-American infants comprised 28% of the found face down group compared to only 12% of the face up or side group. Our data do not support a role for external UAO caused by face positioning directly into the sleep surface in SIDS, but are consistent with internal UAO associated with apnea or gasping before dying of SIDS. PMID- 19291446 TI - Fatality complicating agranulocytosis in the setting of carbimazole therapy. AB - Carbimazole is an antithyroid drug of the thionamide class which is used in the treatment of hyperthyroidism. Rarely, use of this drug may be associated with the development of agranulocytosis and neutropenia with the consequent risk of sepsis. We present the case of a 50-year-old female who developed rapidly progressive Pseudomonas aeruginosa septicemia in the setting of panycytopenia approximately 4 weeks following the commencement of carbimazole therapy for Graves' disease. She died shortly after presentation to hospital and the case was referred to the coroner as the death was unexpected and the clinical course was considered unusual. Relatively sudden, unexpected deaths resulting from hematological/endocrine causes are uncommonly encountered by forensic pathologists and this case serves to illustrate the enduring value of the autopsy in providing important clinicopathological correlation to clinicians. PMID- 19291445 TI - Sudden unexpected death in a toddler with Williams syndrome. AB - We report a toddler with Williams syndrome (WS) complicated by congenital heart disease and a seizure-like episode who died suddenly several hours after uneventful cardiac catheterization. The cardiovascular anomalies and their association with sudden death as well as the clinical neurological and neuropathologic abnormalities identified in WS are delineated and discussed. PMID- 19291447 TI - Small bowel perforation and fatal peritonitis following a fall in a 21-month-old child. AB - The authors report a case of fatal peritonitis due to small bowel perforation in a 21-month-old female child. Necropsy excluded natural disease and a thorough Coronial investigation concluded that an accidental fall onto a "doorstop" caused the bowel injury. The investigative findings are presented; the discussion address issues of diagnosis and causation/mechanism of injury. PMID- 19291448 TI - DNA reviews: DNA identification following CBRN incidents. AB - Chemical, biological, radioactive, or nuclear (CBRN) incidents can occur due to accident or deliberate action, and may result in substantial loss of life. Whatever the cause, the requirement for identification of the deceased may necessitate the removal of contaminated samples to a DNA laboratory for processing. This review looks at the potential types of CBRN that may result in the requirement for DNA identification of the deceased and investigates the potential risks and difficulties associated with processing samples of this type. PMID- 19291450 TI - University Institute of Legal Medicine in Berlin celebrates its 175th anniversary. AB - The University Institute of Legal Medicine of the University Hospital Charite in Berlin is the oldest faculty in Germany. On February 11, 1833 it was founded as a "Praktische Unterrichtsanstalt fur Staatsarzneikunde (Practice-oriented School of Public Health and Medicine)" at the Friedrich-Wilhelms-Universitat (founded in 1810) and in 1886 it received its own building in Berlin-Mitte. Starting with Wilhelm Wagner, Johann Ludwig Casper, Fritz Strassmann, and their successors established the Berlin School of Legal Medicine, which has been playing an important role for the subject's progress in the German-speaking area until today. Regardless of the accepted scientific achievements, the institute fell victim to the Berlin policy of austerity after 2003 leading to staff reduction and the closure of the historical location. Since the new appointment to the chair in 2007 and the junction of the subject in Berlin-Moabit there is now the chance for a new impetus of forensic medicine in the German capital. PMID- 19291451 TI - The advantage of a medical degree. PMID- 19291452 TI - Hyponatremia at autopsy: an analysis of etiologic mechanisms and their possible significance. AB - While electrolyte measurements after death may be confounded by a number of variables, vitreous humor sodium tends to remain stable for some time, enabling correlation between ante- and postmortem levels. Review of natural and unnatural causes of reduced vitreous humor sodium levels at autopsy was undertaken to demonstrate the range of diseases that may result in this finding. Natural diseases affecting the vasopressin-renin-angiotensin axis may cause reduction in sodium levels with associated hypovolemia, euvolemia, and hypervolemia. Low sodium measurements may also occur with redistribution of water, and artefactually when there are underlying lipid and protein disorders. Unnatural causes of hyponatremia at autopsy include water intoxication from psychogenic polydipsia, environmental polydipsia, ingestion of dilute infant formulas, beer potomania, endurance exercise, fresh water immersion (including water births) and iatrogenic causes including drug and parenteral fluid administration, and surgical irrigation. A knowledge of the range of conditions that may result in lowered postmortem sodium levels will help to exclude or confirm certain diseases at autopsy. In addition, significant vitreous hyponatremia may be a useful finding to help clarify mechanisms of unnatural deaths. PMID- 19291453 TI - Forensic pathological evaluation of injury severity and fatal outcome in traffic accidents: five illustrative autopsy cases of clinically unexpected death. AB - In forensic casework, investigation of injury severity in traffic accidents is important for evaluating the mortality, occasionally in terms of the adequacy of clinical management. The present study evaluated 5 cases of clinically unexpected delayed collapse followed by death using the abbreviated injury scale (AIS), injury severity score (ISS), and a clinical trauma care method (trauma and injury severity score, TRISS). In these cases, major injury (AIS = 3-5) was found in the head, chest and/or abdomen at autopsy, and ISS was estimated to be 11-45 (serious to critical but not incompatible with life). By the TRISS method, the probability of survival (P (s)) was estimated to be >0.5 for all cases (0.60-0.99), suggesting that these were preventable deaths. However, the present cases showed several common features: (a) fatality due to closed injury/-ies to the thoracic and/or abdominal viscera, (b) alert and poor symptoms/clinical signs, and (c) poor positive findings in diagnostic imaging at early times after injury, and (d) complications of other evident injuries, suggesting difficulties in the clinical diagnosis of potentially fatal injuries, but (e) possibly predictable fatal injury when typical patterns of traffic accident injury were considered. PMID- 19291454 TI - Non-traumatic causes of perianal hemorrhage and excoriation in the young. AB - The discovery at autopsy or at a death scene of fresh perianal hemorrhage and/or cutaneous excoriation in a young child is always of concern as this raises the possibility of inflicted injury. Three cases are reported where perianal bleeding and excoriation that were initially considered due to possible sexual assault were subsequently found to be of a non-suspicious nature. Case 1: A previously well 18-month-old boy was accidentally hanged. Fresh perianal hemorrhage that had raised the possibility of sexual assault was subsequently shown to be due to perineal streptococcal dermatitis. Case 2: A 2-year-old girl vomited and then stopped breathing. Fresh blood at the anus, that was also thought to be patulous, raised suspicions of sexual assault. At autopsy, however, bleeding around the normally configured anus was due to a midgut volvulus associated with intestinal malrotation. Case 3: A 21-month-old girl was found dead in her cot. Sexual abuse was suspected when lacerations were allegedly found around her anus. These were, however, due to skin lesions from her previously diagnosed ectodermal dysplasia clefting syndrome. Death was due to upper airway obstruction from acute and chronic inflammation. There was no evidence of anogenital trauma or sexual assault in any of the cases. Although inflicted traumatic causes of perianal hemorrhage and excoriation must be judiciously sought in the young, the current cases demonstrate that organic etiologies must also be considered. Perianal infections, congenital malformations of the mesentery and intestines, and inherited disorders of the skin may all produce findings that may initially suggest that sexual assault has occurred. Careful examination with appropriate photography and/or microbiological testing are required. PMID- 19291455 TI - Characterization of two unusual allele variants at the STR locus ACTBP2 (SE33). AB - Two uncommon allelic variants have been observed at the locus ACTBP2 (SE33) and both were located far outside the ladder range of commercially available typing kits. The short variant showed a 60-bp deletion upstream of the 5'-flanking region with a typical type I repeat structure, which actually would have to be assigned as allele 16. The long allele revealed a typical type III sequence structure that has to be designated as allele 41. PMID- 19291456 TI - Failure of apnea monitoring during bed-sharing. AB - We report the case of an infant found dead in bed with an adult while attached to a respiratory monitor sensor. We experimentally re-created a bed-sharing situation with an infant mannequin to demonstrate that a Graseby home apnea monitor continues to sense adult breathing and movement in the absence of infant breathing when used in this sleep situation. PMID- 19291457 TI - Methadone and methadone metabolites in postmortem specimens. AB - We have determined drug/metabolite concentrations and ratios of methadone (METH) to two of its metabolites (EDDP, 2-ethylidene-1, 5-dimethyl-3, 3 diphenylpyrrolidine; and EMDP, 2-ethyl-5-methyl-3,3-diphenylpyrroline) in postmortem peripheral blood and liver tissue by liquid chromatography/tandem mass spectrometry. The assays employed deuterated internal standards and multiple reaction monitoring (MRM) techniques. The assay linear range was 0.01-2.0 mg/l for each analyte. METH, EDDP, and EMDP were determined in liver and peripheral blood from 46 methadone-positive cases. METH and EDDP were detected in all specimens, whether blood or liver. EMDP was detected, only in liver, and only 17 cases, at concentrations much lower than those of EDDP. Concentrations of METH and EDDP in blood and liver from EMDP-positive cases were in ranges higher than, but overlapping with, concentrations in blood and liver from EMDP-negative cases. These data suggest that although METH is readily demethylated and cyclized to EDDP, in vivo, conversion to EMDP may be less efficient and its accumulation in postmortem tissues may be highly individual. PMID- 19291458 TI - Postmortem serum nitrogen compounds and C-reactive protein levels with special regard to investigation of fatal hyperthermia. AB - The present study analyzed serum levels of urea nitrogen (UN), creatinine (Cr), and C-reactive protein (CRP), which are very stable during the early postmortem period, for investigation of the cause of death with special regard to hyperthermia (heat stroke) in serial medico-legal autopsy cases (n = 429), excluding fatal injury, intoxication, and fire fatality. In this series, mechanical asphyxiation, drowning, and sudden cardiac death cases (n = 56, n = 43, and n = 212, respectively) usually showed low levels within postmortem reference ranges for these serum markers, although UN and CRP levels were mildly elevated in cases of sudden cardiac death and cerebrovascular stroke. There were concomitant significant elevations in serum levels of UN (>50 mg/dL), Cr (>2 mg/dL), and CRP (>2 mg/dL) for chronic renal failure, gastrointestinal bleeding, pneumonia, and hypothermia (cold exposure). UN and CRP were especially high for chronic renal failure and pneumonia, respectively. However, hyperthermia cases showed an isolated elevation in the serum Cr level, suggesting an influence of systemic skeletal muscle damage. These serum markers may be practically useful for postmortem investigation of death due to hyperthermia (heat stroke), for which specific pathological and toxicological evidence may not be available. PMID- 19291459 TI - Investigation on the reliability of determining sex from the human os zygomaticum. AB - The diagnosis of sex using the skull is based mainly on morphological findings. The differential expression of these characteristics is described almost identically in the relevant literature. The descriptions date back to the 18th century and stem from observations of inhomogenous investigations, partly with very small samples. Therefore, it is necessary to re-assess the significance of each characteristic. The objective of the present study is to determine the significance that can be attributed to different morphological characteristics and metric data of the os zygomaticum used in sex diagnostics from a current point of view. A collection of 138 recent human skulls was investigated. Both examination of the morphological characteristics as well as metric analysis of the cheekbone in the test group revealed that both methods were suitable for differentiating sex, but due to their variability they cannot be regarded as sufficiently reliable. PMID- 19291460 TI - Histologic dating of bruises in moribund infants and young children. AB - It is generally held that leukocytes are found within bruised subcutaneous tissues within 4-12 h of injury as part of a standard cellular response to trauma. As a corollary, the absence of leukocytes is often cited as evidence of more recent injury. To investigate how long after injury it may be before a leukocyte response occurs selected bruises from three children aged 27, 11, and 3 months, respectively, were examined microscopically. All of the children had sustained lethal head trauma, with survival on life-support equipment for some time in hospital, and with bruises of at least 24-h duration confirmed by medical evaluation (at 30, 44, and 79 h from the time of initial medical evaluation to death). Histologic examination of selected lesions in all three cases revealed extravasation of red blood cells within subcutaneous tissues, but no leukocyte infiltration or other cellular reaction. Other bruises in these children exhibited a standard inflammatory response. This study has shown that selected bruises in three children were present for at least 30 h without a leukocyte infiltrate. Caution should, therefore, be exercised in assigning too rigid a time course to bruising in infants and young children based on a lack of a vital reaction, as the absence of leukocytes within soft tissues of bruised skin in these cases may not necessarily indicate that the injuries are recent. Variability in tissue response may also occur in different bruises in the same individual. Whether severe craniocerebral trauma played a role in delaying the cellular response in these particular injuries is unclear. PMID- 19291462 TI - DNA reviews: predicting phenotype. AB - The prediction of an individuals physical appearance from small biological samples, such as those collected from crime scenes may still sound like science fiction, but how close are we to achieving this goal? This review provides a brief introduction to the areas under investigation for direct and indirect phenotypic inference from DNA alone and suggests some sources of further reading for those interested in gaining a more in-depth knowledge of this complex subject. PMID- 19291463 TI - Guidelines for the effective conduct of mass burials following mass disasters: post-Asian tsunami disaster experience in retrospect. AB - The frequency of mass disasters is increasing, demanding actions that deal with these promptly and effectively to secure human interests. An undeniable and inevitable reality of any mass disaster is the massive number of fatalities, which will give rise to a further chain of events ranging from the recovery of the deceased, to their transport, storage, identification and, finally, disposal. Past experience has shown that traditional human disposal methods should be redesigned according to the requisites of mass fatality scenarios, and it has been proven that a proper mass burial is by far the most appropriate and standard method for disposal of the dead due to mass disasters as it takes all its practical issues into consideration. A mass burial can be defined as burying more than one deceased of a single or related incident in a single grave or multiple graves simultaneously or separately within a restricted time period in a single or multiple burial sites located within an identified geographical area. In the present context, it is an utmost necessity that we develop uniform detailed guidelines for the proper conduct of mass burials that provide the deceased with all due respect to human dignity, as this will enable these guidelines to be incorporated into future national mass disaster management schemes as an integral component. PMID- 19291464 TI - Diagnostic yield and characteristic features in a series of decomposed bodies subject to coronial autopsy. AB - A study was undertaken of 629 cases autopsied at Forensic Science SA, Adelaide, Australia over a 10-year period from 1994 to 2004 where significant decomposition had been documented. The age range of victims was 10 months to 92 years (mean 51.5 +/- 18.1 years). There were 498 males (79%) and 131 females (21%) (M:F = 3.8:1). Deaths in 289 of the 629 cases (46%) were due to natural causes, with 179 suicides (28%), 83 accidents (13%), and 36 homicides (6%). The cause of death was ascertained in 89% of cases and the manner of death in 93% of cases. In 35 cases (6%) both the cause and manner could not be determined. Numbers of suicides were higher in younger age groups while deaths from organic illnesses predominated in later life. The number of cases in summer was significantly greater than in winter. Despite technical difficulties in handling decomposed bodies, and artifactual alteration of tissue structure and microscopic features, the autopsy was still a useful diagnostic exercise. While it is likely that more information may be gleaned from fresh bodies in perfectly preserved states, decomposed bodies may reveal significant anatomical and pathological features that enable both the cause and manner of death to be established. PMID- 19291465 TI - Femur, rib, and tooth sample collection for DNA analysis in disaster victim identification (DVI) : a method to minimize contamination risk. AB - Although much literature is available on DNA extraction from tissue samples to obtain the best possible genotyping results, to the best of our knowledge no written recommendations exist on how to excise or extract bone and tooth samples from a victim to facilitate this. Because the possibility of cross-contamination is high, especially when excising numerous samples under disaster conditions, it is important to minimize this risk and to keep samples in optimum condition. In this paper a standard operating procedure is proposed for collection of femur, rib, and tooth samples to aid victim identification both after mass disasters and in (single) forensic investigations. PMID- 19291466 TI - Decomposition and insect succession on cadavers inside a vehicle environment. AB - This study presents differences in rate of decomposition and insect succession between exposed carcasses on the soil surface and those enclosed within a vehicle following carbon monoxide (CO) poisoning. Nine 45-kg pigs were used as models for human decomposition. Six animals were sacrificed by CO gas, half of which were placed within the driver's side of separate enclosed vehicles and half were placed under scavenger-proof cages on the soil surface. A further three animals were sacrificed by captive headbolt and placed under scavenger proof cages on the soil surface. The pattern of insect succession and rate of decomposition were similar between surface carcasses within trials regardless of the mode of death. Progression through the physical stages of decomposition was 3-4 days faster in the enclosed vehicle due to higher temperatures there compared to external ambient temperatures. Patterns of insect succession also differed between the vehicle and surface treatments. Carcass attendance by representatives of the Calliphoridae was delayed within the vehicle environment by 16-18 h, while oviposition was not observed until 24-28 h following death. In contrast, attendance by Calliphoridae at surface carcasses occurred within 1 h of death, and oviposition occurred within 6-8 h of death. Typical patterns of insect succession on the carcasses were also altered. Carcass attendance by representatives of the Coleoptera occurred during the bloat stage of decomposition at surface carcasses but was delayed until the onset of wet decomposition (as defined by carcass deflation and breakage of the skin) within the vehicle environment. This study provides baseline data outlining the decomposition patterns of a carcass enclosed within a vehicle following CO poisoning in Western Australia. Understanding how variations in decomposition situations impact on the rate of decomposition and patterns of insect succession is essential to obtaining an accurate estimate of minimum post-mortem interval (PMI). PMID- 19291467 TI - The biomechanical modelling of non-ballistic skin wounding: blunt-force injury. AB - Knowledge of the biomechanical dynamics of blunt force trauma is indispensable for forensic reconstruction of a wounding event. In this study, we describe and interpret wound features on a synthetic skin model under defined laboratory conditions. To simulate skin and the sub-dermal tissues we used open-celled polyurethane sponge (foam), covered by a silicone layer. A drop tube device with three tube lengths (300, 400, and 500 mm), each secured to a weighted steel scaffold and into which a round, 5-kg Federal dumbbell of length 180 mm and diameter 8 cm was placed delivered blows of known impact. To calculate energy and velocity at impact the experimental set-up was replicated using rigid-body dynamics and motion simulation software. We soaked each foam square in 500 mL water, until fully saturated, immediately before placing it beneath the drop tube. We then recorded and classified both external and internal lacerations. The association between external wounding rates and the explanatory variables sponge type, sponge thickness, and height were investigated using Poisson regression. Tears (lacerations) of the silicone skin layer resembled linear lacerations seen in the clinical literature and resulted from only 48.6% of impacts. Poisson regression showed there was no significant difference between the rate of external wounding for different sponge types (P = 0.294) or different drop heights (P = 0.276). Most impacts produced "internal wounds" or subsurface cavitation (96%). There were four internal "wound" types; Y-shape (53%), linear (25%), stellate (16%), and double crescent (6%). The two-way interaction height by sponge type was statistically significant in the analysis of variance model (P = 0.035). The other two-way interactions; height by thickness and sponge type by thickness, were also bordering on statistical significance (P = 0.061 and P = 0.071, respectively). The observation that external wounds were present for less than half of impacts only, but that nearly all impacts resulted in internal wounds, might explain the observed haematoma formation and contusions so often associated with blunt-force injuries. Our study also confirms the key role of hydrodynamic pressure changes in the actual tearing of subcutaneous tissue. At the moment and site of impact, transferred kinetic energy creates a region of high pressure on the fluid inside the tissue. As a result of the incompressibility of the fluid, this will be displaced away from the impact at a rate that depends on the velocity (or kinetic energy) of impact and the permeability and stiffness of the polymeric foam and skin layer. PMID- 19291468 TI - Sudden infant death while awake. AB - Epidemiologic data suggest that SIDS is related to the sleep state, but exiguous literature has addressed infants who had been awake at the time of sudden catastrophic deterioration and subsequent death. The aims of this study are to: (1) Report five infants who were awake at the onset of the lethal event, and (2) Discuss potential lethal pathophysiological events that may lead to these circumstances. The demographic and pathologic profiles of these cases are similar to SIDS. Altered responses to severe hypotension, bradycardia, and apnea, perhaps elicited by aspiration and mediated by cerebellar and vestibular structures, might be involved in the pathogenesis of these deaths. Comprehensive medical history review, investigation of the circumstances of death, thorough postmortem examination with ancillary studies, and preservation of tissues for gene testing, are crucial to explaining these deaths. Careful attention should be given to the awake or sleep state immediately prior to the sudden clinical collapse, and death of infants; those who were awake should be reported to enhance understanding of this phenomenon. PMID- 19291469 TI - Fatal bupropion overdose with post mortem blood concentrations. AB - We report five cases of fatal bupropion overdose with post mortem bupropion concentrations ranging from 3.1 to >20 mg/L. Four patients had ingested a sustained-release formulation of bupropion and had evidence of pill "bodies" in their stomach with significantly elevated blood bupropion concentrations. The pills found in these patients may represent the residual matrix/shell with significant portions of the actual bupropion released and absorbed by the patients. PMID- 19291470 TI - Deaths in a head-down position: a case report and review of the literature. AB - Deaths in a head-down position, mostly accidental events, are rare deaths in which a victim is found in an inverted body posture, with marked congestion of dependent body parts and no definite pathoanatomical cause of death. Such an exclusion diagnosis can only be put forward after elimination of other possible causes of death, following a scene investigation, medical record review, complete autopsy, and toxicological analysis. Particular attention should be taken not to confuse deaths by head-down position from the more usual positional asphyxia. We here report the case of an 82-year-old woman found stuck in the railings of the staircase leading to her house, her body freely suspended downward below the stairs. Death was finally attributed to a head-down position when correlating the autopsy findings with elements from the scene. Case reports of death by head-down position in the literature are reviewed. Furthermore, results from human experimental studies of head-down tilt are compiled. This systematic review allows an interesting insight into the physiopathology of those deaths, pointing more toward heart failure than to an asphyxial phenomenon per se. PMID- 19291471 TI - The mysterious death of America's richest woman in 1917, Mary Lil' (RIP). AB - Suspicious circumstances surrounding the death of wealthy socialite Mary Lily "Lil'" Kenan Flagler Bingham in 1917 prompted a secret autopsy conducted by renowned physicians at the request of her relatives. Mrs Bingham, who was formerly married to railroad magnate, Henry Flagler, was among America's richest people at the time of her death. Mary Lil's untimely death occurred approximately six months after her marriage to Judge Robert "Bob" Worth Bingham, a college boyfriend whom she married three years after Flagler's death. In order to establish Mary Lil's medical condition prior to her death, a book about Mary Lil's life written by Pulitzer Prize author, David Chandler and articles from the New York Times; The Louisville Courier Journal, a Kentucky newspaper; and Mary Lil's hometown newspaper, The Morning Star, in Wilmington, North Carolina were examined. Although a team of medical experts performed a secret autopsy following her death and released their findings to her family, the examination and tests results remain undisclosed to the public. PMID- 19291474 TI - Retrospective diagnoses of conditions affecting historical individuals. PMID- 19291472 TI - Cancer and forensic microsatellites. AB - Although not directly related, circumstances do occur in forensic investigations whereby cancer studies and forensic science cross paths. This review takes a look at the circumstances under which this may occur, and investigates some potential problems that can arise when tumor tissue is submitted for DNA profile analysis. A background to the underlying molecular biology of tumors is described, highlighting the genetic instabilities that are observed in DNA sequences of similar or identical primary structure to the short tandem repeat markers used in forensic DNA profiling kits. PMID- 19291475 TI - Forensic pathology and problems in determining cause of death. PMID- 19291476 TI - Forensic veterinary medicine: a rapidly evolving discipline. AB - Forensic veterinary medicine is of increasing importance. Legal and other cases in which a veterinary input is likely to be required include those relating to unexpected death of animals, welfare, abuse, and breaches of conservation law. The principles of investigation closely follow those used in human forensic medicine, but there are some important differences. PMID- 19291477 TI - Trends in sudden infant death syndrome in Australia from 1980 to 2002. AB - Analysis was undertaken of trends in sudden infant death syndrome (SIDS) in Australia from 1980 to 2002 using Australian Bureau of Statistics data. The results showed a decline in the SIDS mortality rate from an average of 195.6 deaths per 100,000 live births in the period 1980-1990 to an average of 96.5 deaths per 100,000 live births in the period 1991-1996 and 51.7 deaths per 100,000 live births in the period 1997-2002. The Poisson regression coefficients for SIDS fitted to the rates at the Australia level indicated that the Reduce the Risks (RTR) campaign led to a significant decline of almost 40% when contrasting the pre- and post-campaign periods. Despite recent suggestions that the fall in SIDS rate has been due to natural variations in incidence, the data clearly show that the decline in SIDS rates coincided almost immediately with the introduction of the RTR campaign and has been sustained over time. There was no evidence of diagnostic transfer or of a postponement of death from infancy to early childhood years. Unless this dramatic fall was caused by as yet undetected factors, the campaign is the only plausible explanation for the markedly reduced SIDS rate in Australia. PMID- 19291478 TI - Mandibular morphology as an indicator of human subadult age: geometric morphometric approaches. AB - Methods for skeletal identification have a long history in physical and forensic anthropology. Recent literature demonstrates that new methods are constantly being developed, concurrent with refinements to those already commonly employed. The present study concerns the application of geometric morphometrics to assess the potential of mandibular morphology as a developmental marker for estimating age at death in subadult human skeletal remains. The sample comprises 79 known age and sex subadult individuals of South African Bantu and African American origin; 38 bilateral three-dimensional landmarks were designed and acquired using a portable digitizer. Linear regression was used to predict age using the multivariate descriptors of mandible size and shape based on configurations of three-dimensional landmarks. Our results show that the mandible can be used to predict age in the subadult skeleton with accuracy comparable to standards based on the dentition (standard error rates are between +/-1.3 and +/-3.0 years). These results closely parallel our previous study using the linear measurement of ramus height, but suggest that geometric morphometrics may be slightly more accurate when adolescents are included in the sample. PMID- 19291479 TI - The postmortem distribution of ketone bodies between blood, vitreous humor, spinal fluid, and urine. AB - The distribution of the ketone bodies: acetone, acetoacetate, and D-beta hydroxybutyrate, between blood, vitreous humor, spinal fluid, and urine was examined in 105 medico-legal autopsies. The ketone body concentration in the body fluids was determinated by head-space gas chromatography. The correlation between blood and the body fluids could be described with regression lines on the logarithmic-transformed results. The correlation is dependent on the ketone body concentration. The ketone bodies in spinal fluid show the best correlation to blood, followed by vitreous humor, and last urine. The concentration dependence in spinal fluid is mainly due to ketone bodies being metabolized in the brain. The human brain utilizes ketone bodies during normal nutritional state. In vitreous humor, the dependence is mainly due to protein bindings of acetoacetate and beta-hydroxybutyrate in blood and the difference in dry matter between blood and vitreous humor. PMID- 19291480 TI - Characterization of cardiomyopathy cases at a forensic institute in the period 1992-2006 and perspectives for screening. AB - We surveyed our data over a 15-year period to determine the prevalence of cardiomyopathy diagnoses in our database and characterized the cases in regards to demographic and clinicopathological data. Furthermore we evaluated implications for screening. The data were acquired through our computerized database containing autopsy reports. Only cases with typical anatomical and histological features were included. The total number of autopsies in the studied period was 7,185 of which 32 suited the following criteria: 14 arrythmogenic right ventricular cardiomyopathies (ARVC), 8 hypertrophic cardiomyopathies (HCM), 8 dilated cardiomyopathies (DCM) and 2 other cardiomyopathies. Symptoms of cardiac disease were present in 8 ARVC, 3 HCM and 7 DCM cases. Symptoms of cardiac disease, including suspected sudden cardiac deaths, were present in the families of 5 cases. In two of these families the illnesses were discovered as a result of testing after autopsy. We underscore that these diseases are difficult to diagnose as the presenting symptoms are elusive. We found the circumstances around death to be extremely varied, and found that new hereditary cases could be discovered after autopsy. This places an intriguing perspective on the integration of standardized screening protocols with participation from forensic institutes. PMID- 19291481 TI - Pseudo-gunshot wound injury from perforating rib fracture: a cautionary case report. AB - We report a case of a young woman who was found unresponsive in a garbage dumpster beneath the balcony of her 9th floor apartment residence. Initial investigations by the police and coroner raised concerns regarding the circumstances of the death. Initial examination of the body, revealed a single penetrating injury on the chest with a wide abrasion collar--the injury pattern was similar to a gunshot entry wound or a shored exit wound. Autopsy revealed the abraded skin perforation along with major chest injuries with bruising of the chest wall, extensive comminuted rib fractures, and pleural and lung lacerations. These injuries were consistent with the effects of a fatal descent from height. The penetrating injury with the abrasion collar was due to a fractured end of a rib protruding through the skin mimicking a gunshot wound. The involvement of a firearm was further excluded by postmortem radiography (no projectile in situ), detailed external examination of the clothing, and scene investigation revealing that the decedent had descended from a balcony into the dumpster. This case underscores the concept that all round penetrating wounds with abrasion collars are not gunshot injuries. PMID- 19291482 TI - Fatal migration of vena caval filters. AB - Inferior vena caval filters, which are used to prevent pulmonary thromboemboli, may migrate to the right ventricle of the heart. We describe two deaths after the migration of IVC filters. In neither was the filter incorrectly inserted or deployed. In each instance, a deep pelvic/leg vein thromboembolus was associated with the dislodged filter. When a thromboembolism becomes entrapped in the filter, the surface area increases and the previous low-resistance filter becomes obstructed or partially obstructed. The force of the blood flow on this entangled wire/clot may cause the filter/clot to embolize to the heart. Even though one may be tempted to certify the death as due to the failure of the filter, death likely would have occurred if the filter were not in place. If the deep venous thrombus is caused by a disease (not an injury or other medical complication), the manner of these deaths is best certified as natural. PMID- 19291483 TI - Fall downstairs: accident, homicide or natural death? AB - In homicide cases a clear anatomical or toxicological cause of death is normally evident. In the following a case is described where, without clear anatomical or toxicological cause of death, the husband of the deceased-a trained anaesthesiologist-was charged with murder. Suspicion was raised since the autopsy findings did not correspond to a fatal fall downstairs but instead could be definitely ruled out as cause of death, since intracranial bleedings and cerebral contusions, the leading causes of death in fatal falls downstairs, were missing. Further suspicion was raised since electrocardiograms (ECGs), previously stated by the accused to have been recorded personally from his wife during cardiopulmonary resuscitation (CPR), were obviously faked. Additionally, an ampule of succinylcholine was missing from his emergency case. In a trial lasting more than three quarters of a year with several witnesses and experts heard by the court, the cause of death remained unclear. However, a natural cause of death and a fatal fall downstairs were ruled out. Subsequently the husband admitted during the trial to have killed his wife without elaborating on the circumstances for legal reasons. He was sentenced to prolonged imprisonment. Of special importance was that, based on the knowledge of typical autopsy findings in fatal falls downstairs, a lethal fall downstairs could be ruled out. Therefore even negative autopsy findings are of great forensic importance. PMID- 19291484 TI - DNA reviews: low level DNA profiling. AB - Low copy number (LCN) DNA profiling has recently been scrutinized in the United Kingdom following the comments of Mr Justice Weir made during the trial of suspected terrorist Sean Hoey. Mr Hoey was acquitted of all charges related to the Omagh bombing of 1998, following the inadmissibility of key DNA evidence during this trial. The Association of Chief Police Officers and Crown Prosecution Service, initially suspended the use of this technique, but quickly reinstated its use following an internal enquiry. This review describes the low copy number technique and the sample types that are now routinely collected from suspects, victims, and crime scenes for examination by this method. PMID- 19291485 TI - Embryology, normal anatomy, and imaging techniques of the hyoid and larynx with respect to forensic purposes: a review article. AB - Experiences with pre- and postmortem imaging in a forensic setting create more and more interest. If autopsy is permitted, forensic pathologists perform careful examination of the neck structures, which plays an important role as this is decisive in the diagnostics of compressive neck injury. Primary tools are important: forensic pathologists and radiologists are supposed to be aware of the complex anatomy of the neck, especially the laryngeal region, to interprete their findings at autopsy and after imaging. It is of great interest whether CT and MR imaging techniques would be useful in comparing findings of autopsy and document findings if autopsy is refused. In the light of this, the embryology, anatomy and a review of imaging techniques of the neck will be highlighted in this article, with special attention to the value of updated imaging techniques of the larynx for forensic purposes in living people and postmortem. PMID- 19291490 TI - Relationships among perceived burden, depressive cognitions, resourcefulness, and quality of life in female relatives of seriously mentally ill adults. AB - Providing care and support to a seriously mentally ill (SMI) family member can have deleterious effects on one's health and quality of life. This study explored relationships among perceived burden, depressive cognitions, resourcefulness, and quality of life in 60 African-American and Caucasian women family members of SMI adults. Caucasians reported greater burden than African-Americans; the groups were similar in depressive cognitions, resourcefulness, and quality of life. In Caucasians and African-Americans, burden correlated with depressive cognitions and both correlated with poorer mental health. In African-Americans, burden also correlated with lower personal resourcefulness and both correlated with poorer mental health. The findings suggest a mediating role by depressive cognitions for both groups and by resourcefulness in African-Americans. Thus, both groups of women may benefit from positive thinking while African-Americans also may benefit from learning personal resourcefulness skills. PMID- 19291491 TI - Raising adults as children? A report on milieu therapy in a psychiatric ward in Norway. AB - Milieu therapy is widely used as a therapeutic approach in psychiatric wards in the Nordic countries, but few studies exist that report on what practices a milieu therapy approach implies as seen from an ethnographic perspective. Therefore, there is a need to obtain insight into how milieu therapy unfolds in a psychiatric ward setting. The present ethnographic study aims to explore this in a locked-up psychiatric ward that was tied to a psychodynamic-oriented milieu therapy approach. Metaphors from traditional nuclear family life were widely used. Patients were often understood as harmed children and were taught self management skills; the staff aimed at providing a caring atmosphere; and the patients seemed to behave, sometimes, in a childlike manner. In a Foucaultian framework, milieu therapy can be seen as a therapeutic normalization technique used to produce self-governing individuals. Milieu therapy "raises" patients in order to transform patients' odd behaviour and nonconforming lifestyles. We see this "raising children" approach as a type of intervention that nicely connects to the national policy of normalization and integration politics towards persons with psychiatric diagnoses. PMID- 19291492 TI - Crisis prevention management: a program to reduce the use of seclusion and restraint in an inpatient mental health setting. AB - The use of seclusion and restraint in the treatment of mentally ill patients is a highly controversial and potentially dangerous practice. A group of direct care psychiatric nurses in a large urban teaching hospital created an evidenced-based performance improvement program that resulted in a decrease in the use of seclusion and restraint. No additional funds were required to develop this program. The public health prevention model was the framework utilized. Early results show a 75% reduction in the use of seclusion and restraint with no increase in patient or staff injuries since its implementation. PMID- 19291493 TI - Electronic screening for mental health in rural primary care: implementation. AB - The goals of this study were to develop a computer-based electronic screening tool (eScreening) and determine the feasibility of implementing eScreening for rural users of primary care. This descriptive pilot adapted existing screening measures for depression and alcohol abuse to a portable computer-based format and examined the feasibility of its adoption and use. This was a three-step design using convenience samples for (1) a focus group with providers, (2) usability testing with selected rural patients using the computerized touch screen, and (3) implementing the touch screen platform with a small sample in primary care to determine feasibility. This paper reports on Phase III, which assessed consumer response to eScreening. PMID- 19291494 TI - Exploring male and female patients' experiences of psychiatric hospital care: a critical analysis of the literature. AB - In a critical analysis of the literature, this paper aimed to explore male and female patients' experiences of psychiatric hospital care. A method for a systematic review of research was used and five articles fitting the inclusion criteria were found. Three themes emerged in the analysis: "Treatment specifically related to women," "To keep a facade," and "Single-sex or mixed ward." The findings were explored by highlighting some knowledge gaps and gender bias. This paper can be used in the planning of psychiatric wards, give incitements to develop strategies for caring, and promote nurses' gender sensitivity. PMID- 19291495 TI - Strategies nurses use to overcome medication refusal by involuntary psychiatric patients. AB - Nurses seem to play an important role in assisting involuntarily hospitalized psychiatric patients to accept medication. The initial aim of this study was to develop a theoretical understanding of strategies nurses use to overcome medication non-acceptance in involuntary psychiatric patients using a grounded theory approach. Interviews (n = 17) were conducted with psychiatric nurses in inpatient settings. Data analysis using the constant comparative method and validation by nursing literature identified four key themes: Engagement, formulating a therapeutic relationship, finding out why, and persistently trying everything. These themes were frequently encountered in descriptions of getting patients to take medications. The findings indicate that psychiatric nurses use varied and individualized techniques to convince involuntary patients to take their medications. PMID- 19291496 TI - Effectiveness of cognitive-behavioral therapy on depressive symptomatology, stress and coping strategies among Jordanian university students. AB - The study examined the effectiveness of cognitive behavioral therapy (CBT) with university students suffering from moderate to severe depressive symptoms in Jordan. Eighty-four university students were recruited and assigned randomly to control and intervention groups. Intervention impact was assessed on measures of depressive symptoms, perceived stress, and coping strategies at three time points; baseline, postintervention, and 3-months postintervention. The interventional model used was the Modified Teaching Kids to Cope (MTKC), and the control group received no treatment. Overall, using CBT showed a significant improvement in the outcome measures. At postintervention, students had lower scores on perceived stress, lower depressive symptoms, less use of avoidance coping strategies, and more use of approach coping strategies. The findings are discussed in terms of treatment implications and recommendations for use at academic and health care settings. PMID- 19291497 TI - Deciding to place a relative in long-term care: "we really don't do that". PMID- 19291498 TI - Stigmatizing language with unintended meanings: "persons with mental illness" or "mentally ill persons"? PMID- 19291499 TI - A new paradigm for quality and safety in nursing. PMID- 19291500 TI - Memantine (Namenda, Forest Pharmaceuticals). PMID- 19291502 TI - Community reactions to a syphilis prevention campaign for gay and bisexual men in Los Angeles County. AB - "Stop the Sores" (STS), a humor-based syphilis prevention campaign, was implemented in response to increasing syphilis prevalence among gay and bisexual men in Los Angeles County. In 2004, 564 men completed surveys measuring exposure and reactions to the campaign and syphilis testing. Mean age was 39, and men of color comprised a significant proportion of the sample (46.8%). Most men reported being HIV-negative (79.3%). Overall, 7.8% of the sample reported ever having syphilis; HIV-positive men were six times more likely to report this. Over one half of the sample (58.5%) reported exposure to the campaign. Men reporting any recent unprotected anal sex were twice more likely (than those who did not) to see the campaign. Men of color were twice more likely than White men to report wanting to speak to their friends about it. Finally, 39.1% of men exposed to the campaign reported being tested for syphilis as a result. Factors related to higher likelihood to test for syphilis included HIV seropositive status, any recent unprotected anal insertive sex, recent use of methamphetamine, recent use of "poppers," and recent use of erectile dysfunction drugs. Although STS was somewhat effective, outreach efforts to particular subgroups may need to increase. PMID- 19291503 TI - Influence of changing occlusal support on jaw-closing muscle electromyographic activity in healthy men and women. AB - OBJECTIVES: To test whether changes in occlusal support differentially modulate masseter and anterior temporalis muscle electromyographic (EMG) activity during controlled maximal voluntary clenching. MATERIAL AND METHODS: Forty-seven healthy subjects (32 M and 15 F, 22.9+/-1.3 years) were recruited. Cotton-rolls were used to modify the occlusal contact relations and were positioned on the right, left, or both sides, and either in the molar or premolar regions, i.e. six different occlusal combinations. Surface EMG activity was recorded bilaterally from the masseter and anterior temporalis area and normalized with respect to maximal voluntary clenching in the intercuspal position. Analysis of variance and the paired t-test were used to test the data. RESULTS: Normalized EMG activity was influenced by changes in cotton-roll modified occlusal support, and there were differences between muscles (p<0.001). In general, EMG activity decreased in both muscles when occlusal support was moved from the molar to the premolar region. When occlusal support was moved from bilateral to unilateral contacts, EMG activity in the balancing-side anterior temporalis muscle and in bilateral masseter muscles decreased. Unilateral clenching on the molars, but not on the premolars, was associated with lower EMG activity in the balancing-side masseter and always associated with lower EMG activity in the balancing-side anterior temporalis compared to the working side (p<0.05). CONCLUSIONS: Masseter and anterior temporalis muscles respond differently to changes in occlusal support, which may have implications for stability of the mandible during intense clenching. PMID- 19291508 TI - AL-Base: a visual platform analysis tool for the study of amyloidogenic immunoglobulin light chain sequences. AB - AL-Base, a curated database of human immunoglobulin (Ig) light chain (LC) sequences derived from patients with AL amyloidosis and controls, is described, along with a collection of analytical and graphic tools designed to facilitate their analysis. AL-Base is designed to compile and analyse amyloidogenic Ig LC sequences and to compare their predicted protein sequence and structure to non amyloidogenic LC sequences. Currently, the database contains over 3000 de identified LC nucleotide and amino acid sequences, of which 433 encode monoclonal proteins that were reported to form fibrillar deposits in AL patients. Each sequence is categorised according to germline gene usage, clinical status and sample source. Currently, tools are available to search for sequences by various criteria, to analyse the biochemical properties of the predicted amino acids at each position and to display the results in a graphical fashion. The likelihood that each sequence has evolved through somatic hypermutation can be predicted using an automated binomial or multinomial distribution model. AL-Base is available to the scientific community for research purposes. PMID- 19291509 TI - Biochemical characterisation of amyloid by endomyocardial biopsy. AB - Cardiomyopathy is a major cause of death in patients with systemic amyloidosis. There are several forms of systemic amyloidosis which cause cardiomyopathy and determination of the exact type of amyloid in each affected patient is essential for treatment and determination of prognosis. In this study, we tested the feasibility of determining the type of amyloidosis by biochemical analysis of endomyocardial biopsies. Right ventricular endomyocardial biopsies were obtained from 10 patients with restrictive cardiomyopathy. Three patients had monoclonal protein demonstrated in serum or urine and all three had bone marrow findings consistent with monoclonal gammopathy. Seven patients had isolated cardiomyopathy without evidence of monoclonal gammopathy. A portion of each myocardial biopsy was submitted for histologic evaluation and all demonstrated amyloid by Congo red staining. Each biopsy was analysed biochemically by isolation of amyloid fibrils and the protein characterised by amino acid sequence analysis. Four amyloid isolates were characterised as immunoglobulin light chain proteins. Two specimens obtained from patients with transthyretin (TTR) DNA mutations contained TTR peptides proving the hereditary nature of the disease. Biopsies from four patients without a TTR mutation contained TTR and were consistent with the diagnosis of senile cardiac amyloidosis (SCA). All endomyocardial biopsy specimens that were analysed had sufficient amyloid fibril subunit protein to allow characterisation by amino acid sequence analysis. This methodology is particularly useful in differentiating SCA with TTR amyloid fibrils from immunoglobulin light chain amyloidosis which also occurs in the elderly age group. PMID- 19291510 TI - Ameliorative effect of 1,2-benzenedicarboxylic acid dinonyl ester against amyloid beta peptide-induced neurotoxicity. AB - Amyloid beta peptide (Abeta)-induced oxidative stress may be linked to neurodegenerative disease. Ethanol extracts of Rosa laevigata protected PC12 cells from hydrogen peroxide-induced oxidative stress. (3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyl tetrazolium bromide) reduction assays revealed a significant increase in cell viability when oxidatively stressed PC12 cells were treated with R. laevigata extract. The effect of R. laevigata on oxidative stress-induced cell death was further investigated by lactate dehydrogenase release assays and trypan blue exclusion assays. Administration of 1,2-benzenedicarboxylic acid dinonyl ester from R. laevigata extract to mice infused with Abeta significantly reversed learning and memory impairment in behavioural tests. After behavioural testing, the mice were sacrificed and brains were collected for the examination of lipid peroxidation, catalase activity and acetylcholinesterase (AchE) activity. These results suggest that 1,2-benzenedicarboxylic acid dinonyl ester from R. laevigata extract may be able to reduce Abeta-induced neurotoxicity, possibly by reducing oxidative stress. Therefore, R. laevigata extract may be useful for the prevention of oxidative stress-induced neurodegenerative disorders. PMID- 19291511 TI - Slaughtered aged cattle might be one dietary source exhibiting amyloid enhancing factor activity. AB - It has been shown that experimental murine AA amyloidosis can be enhanced by dietary ingestion of amyloid fibrils, and it is known that systemic AA amyloidosis occasionally develops in aged cattle. In this study, we examined amyloid deposits in renal and muscular tissues simultaneously obtained from slaughtered aged cattle; from both tissues when affected, amyloid-enhancing activity was also investigated. On histopathology, renal amyloid deposition was seen in nine of the 293 cattle with no history of disease, and minute amyloid deposition in muscular tissue was detectable in one of these nine. All these amyloid deposits were immunohistochemically demonstrated to be AA. Extracts, which might contain amyloid fibril fractions, were isolated from renal and muscular tissues in five of these nine cattle. On SDS-PAGE and Western blot analysis, protein bands immunoreactive to anti-AA serum were detected in the kidney fractions obtained from four of the five latter cattle, but no bands were seen in the muscle fractions of any of the five cattle. Amyloid fibril fractions from two cattle were intravenously injected into group of seven experimentally designed mice for induction of AA amyloidosis. All seven mice injected with kidney fraction developed severe AA amyloidosis, whereas only one of the seven mice given muscle fraction showed slight amyloid deposition in the spleen. These data suggest that food products made from aged cattle possess amyloid-enhancing potential. PMID- 19291512 TI - Epidemiological, clinical and genetic study of familial amyloidotic polyneuropathy in Cyprus. AB - OBJECTIVES. To define the incidence and prevalence of familial amyloidotic polyneuropathy (FAP) TTRVal30Met on the island of Cyprus. To study the clinical phenotype and genetic features of FAP TTRVal30Met in the Cypriot population. METHODS: The clinical and neurogenetic databases were used to identify probands with FAP TTRVal30Met and detailed family trees were constructed. Potential carriers of the mutation were identified from the family trees and assessed clinically and genetically. Transthyretin was completely sequenced in patients and potential carriers. RESULTS: Thirty-six patients carrying the TTRVal30Met mutation (one homozygote) from 22 families were identified. On 1 December 2003 the prevalence of FAP was 3.72/100,000 while the incidence is estimated to be 0.69/100,000 per year. The phenotype observed was characteristic for a length dependent sensorimotor and autonomic neuropathy with neuropathic pain. Mean age of onset was 46 years. Penetrance is estimated to be 28% and positive anticipation in the age of onset is found. CONCLUSION: FAP is relatively prevalent in Cyprus which may be considered as another endemic focus of the disease in Europe. The mean age of onset and penetrance is different from the Portuguese and Swedish populations. Understanding the biological factors that determine these differences could potentially lead to therapeutic advances. PMID- 19291513 TI - Discordant expression of familial amyloid polyneuropathy in monozygotic Brazilian twins. AB - Discordant expression of Familial Amyloid Neuropathy (FAP) in monozygotic twins is a rare event. Only five such cases have been described in the literature so far. We report the clinical, neurophysiologic and autonomic findings of Brazilian monozygotic twins discordant for the expression of FAP type I. Twin I first presented symptoms at the age of 21, when his brother was completely asymptomatic. Twin 2 only presented symptoms at the age of 25, almost four years after his brother. Both brothers eventually developed the complete phenotype of FAP type I. The occurrence of monozygotic twins discordant for the expression of FAP type I suggests that other factors beside TTR gene mutations should play an important role in the pathogenesis of this condition. Environmental factors, as well as modifier genetic loci are likely to modulate the expression of FAP type I and the study of cases such as the one presented here may help to identify some of these factors. PMID- 19291514 TI - Amyloidosis, Evans syndrome and management options of lymphoplasmacytic lymphoma. AB - A 77-year-old man presented with Evans syndrome (ES), hard palate thickening, gastrointestinal (GI) hemorrhage, acute myocardial infarction (AMI) and pleural and pericardial effusions. The patient responded well to emergent ES treatment with high-dose steroids and intravenous immunoglobulin. Investigation revealed lymphoplasmacytic lymphoma (LPL) as well as amyloidosis in the hard palate, lymph nodes, and pericardium. Considering his age, non-myelosuppressive agents were administered, with the exception of dose-reduced cyclophosphamide. The patient developed neutropenic fever, atrial fibrillation and subsequently died. This report describes the first LPL patient with ES. LPL is generally an indolent disease. However, as in our patient, it can be life threatening because of its complications. ES contributed to his GI hemorrhage, severe anemia, and thus AMI at the time of presentation. Probable cardiac amyloidosis played a role in the latter phase (i.e. cardiac arrhythmia and hypotension during sepsis). Although rare, the presence of ES and amyloidosis should be investigated diligently in elderly LPL patients. Instead of aggressive myelosuppressive chemotherapy agents, targeted therapies might be considered in these fragile patients. PMID- 19291515 TI - Atraumatic splenic rupture in amyloidosis. AB - BACKGROUND: Splenic involvement in amyloidosis is rather frequent (5-10%). An atraumatic rupture of the affected spleen is however an extremely rare event. We report on a patient with undiagnosed amyloidosis who underwent emergency splenectomy for atraumatic splenic rupture. METHODS: Review of the literature and identification of 31 patients, including our own case report, with atraumatic splenic rupture in amyloidosis. Analysis of the clinical presentation, the surgical management, the nomenclature and definition of predisposing factors of splenic rupture. RESULTS: We identified 15 women and 16 men (mean age 53.3 +/- 12.4 years; median 52, range: 27-82 years) with an atraumatic splenic rupture. Easy skin bruisability and factor X deficiency were detected in four (13%) and five patients (16%), respectively. The diagnosis of splenic rupture was made either by computed tomography (n = 12), ultrasound (n = 5), exploratory laparotomy (n = 9) or autopsy (n = 4). All patients underwent surgery (n = 27) or autopsy (n = 4). Amyloidosis was previously diagnosed in nine patients (29%). In the remaining 22 patients (71%), the atraumatic splenic rupture represented the initial manifestation of amyloidosis. Twenty-five patients (81%) suffered from primary (AL) and four patients (13%) from secondary amyloidosis (AA). In two patients, the type of amyloidosis was not specified. A moderate splenomegaly was a common feature (68%) and the characteristic intraoperative finding was an extended subcapsular hematoma with a limited parenchymal laceration (65%). In five patients with known amyloidosis, the atraumatic splenic rupture was closely associated with autologous stem-cell transplantation (ASCT) (16%). Three patients were suffering from multiple myeloma (10%). A biopsy-proven amyloidotic liver involvement was present in 14 patients (45%), which lead to atraumatic liver rupture in two patients. The splenic rupture related 30-day mortality was 26% (8/31). CONCLUSIONS: Atraumatic splenic rupture in amyloidosis is associated with a high 30-day mortality. It occurs predominantly in patients with previously undiagnosed amyloidosis. A moderate splenomegaly, coagulation abnormalities (easy skin bruisability, factor X deficiency) and treatment of amyloidosis with ASCT are considered predisposing factors for an atraumatic splenic rupture. PMID- 19291516 TI - Concomitant amyloidosis, renal papillary carcinoma and ipsilateral pelvicalyceal urothelial carcinoma in a patient with familial Mediterranean fever. AB - We report a case of a 58-year-old man with a history of long standing familial Mediterranean fever (FMF) and AA amyloidosis, who developed renal papillary carcinoma and renal pelvic urothelial carcinoma simultaneously. Although the association between chronic inflammatory states like FMF and AA amyloidosis has been well established, the relationship between amyloidosis and solid tumors is not defined as clearly. Furthermore, to the best of our knowledge, co-existence of two different types of kidney malignancy with amyloidosis in a patient with FMF has not been reported. Our patient was admitted to hospital with gross hematuria and renal insufficiency. Imaging studies revealed mass lesions in the middle portion of the right kidney. Right radical nephrectomy showed extensive amyloid deposition, co-existing with renal papillary carcinoma and poorly differentiated invasive urothelial carcinoma. PMID- 19291518 TI - Update on integrated biomarkers for assessment of long-term risk of cardiovascular complications in initially healthy subjects and patients with manifest atherosclerosis. AB - Risk stratification for cardiovascular diseases (CVD) remains suboptimal even after the introduction of global risk assessment by various scores. This has prompted the search for additional biomarkers which might help to improve risk stratification. Basically, there are blood biomarkers representing various pathophysiological pathways of atherosclerosis, and markers of subclinical disease. Since inflammatory processes accompany all stages of atherosclerosis, measurement of plasma/serum concentrations of circulating inflammatory biomarkers have received great attention. Such biomarkers can be measured systemically by sensitive assays, and elevated concentrations in the circulation have been shown to be associated with future CVD events. Thus, they might add to the predictive value of the atherogenic lipoprotein phenotype to further improve CVD risk assessment. In addition, several non-invasive imaging techniques are available for which also a predictive value for CVD could be established. However, for most of these biomarkers the clinical utility has not yet been firmly established. This review attempts to give an update on the potential use of biomarkers for risk stratification in initially healthy subjects and patients with manifest, chronic atherosclerosis, particularly focusing on the integrated value of the combination of these markers. PMID- 19291526 TI - Venous thromboembolism (VTE) in patients with cancer: epidemiology and risk factors. AB - Although cancer is recognized as a major risk factor for venous thromboembolism, the exact magnitude of the problem and specific risk factors most strongly associated with the development of VTE is not well defined. Several recent studies have shown that the incidence of VTE is highest in patients who present with metastatic cancer, particularly cancers associated with a high one-year mortality rate, such as pancreatic cancer. The incidence rate of VTE is highest in the first few months after the diagnosis of cancer, and it decreases over time thereafter. For most cancers, it is not clear to what extent undergoing major surgery adds to the already high risk of VTE associated with the presence of the cancer. However, patients with glioma clearly have a very high incidence of VTE soon after they undergo any invasive neurosurgical procedure. Active chemotherapy, the use of erythropoetin agents, and the use of certain anti-cancer therapies such as thalidomide, high-dose steroids, and anti-angiogenic therapy also increase the risk of thrombosis. Similar to patients without cancer, the risk of VTE is higher in patients with coexisting chronic medical illnesses. Development of VTE is clearly associated with decreased survival and this effect is greater among patients initially diagnosed with local or regional stage cancer compared to patients with metastatic cancer. PMID- 19291527 TI - Prognostic models: a methodological framework and review of models for breast cancer. PMID- 19291528 TI - Narratives of desire in mid-age women with and without arousal difficulties. AB - There is controversy about the nature of women's sexual desire. The aim was to explore narrative descriptions of sexual desire among mid-aged women in hopes of clarifying how women define and experience sexual desire, and how these might differ among women with and without female sexual arousal disorder (FSAD). Mid aged women without (age: M = 45, n = 12) and with (age: M = 55, n = 10) FSAD took part in in-depth interviews that invited them to share personal stories of sexual desire. Women also completed the Brief Index of Sexual Functioning and the Female Sexual Function Index (FSFI). Women in both groups described sexual desire in genital, non-genital physical, and in cognitive-emotional terms. Although women with FSAD had low ratings of sexual desire on the FSFI, they could recall recent experiences of desire that did not differ from the control group. Women identified a number of triggers of desire including touch, memories, and partner's responses--the latter of which acted as both a trigger and an inhibitor. Women in the control group were more likely to express conflation about the distinction between desire and arousal. Among the different "objects" of women's desire, most women acknowledged emotional connection as most important. PMID- 19291529 TI - Goal-oriented cognitive rehabilitation for an individual with Mild Cognitive Impairment: behavioural and neuroimaging outcomes. AB - An individual, goal-oriented cognitive rehabilitation intervention was conducted with AB, a 77-year-old woman with MCI. Outcomes were evaluated using a client centred measure of goal achievement and an experimental associative learning task testing a skill practised in the intervention. Pre- and post-intervention fMRI scans were compared to examine changes in brain activation. Following intervention, AB showed improved performance on rehabilitation goals and on the associative learning task. There were significant decreases in brain activation in sensory areas and significant increases in memory-related areas. Both client centred measures and fMRI may assist in detecting clinically-meaningful changes resulting from cognitive rehabilitation interventions. PMID- 19291530 TI - Throwing the baby out with the bathwater: microRNAs have critical roles in health and disease. PMID- 19291531 TI - The clinicopathological relevance of microRNA in normal and malignant haematopoiesis. AB - MicroRNAs (miRNAs) are recently discovered short non-coding RNA molecules that negatively regulate messenger RNA (mRNA) translation to protein. Their discovery heralds a novel mechanism of post-transcriptional gene regulation and has lead to a cascade of studies aimed at identifying how miRNA dysregulation may contribute to disease. Recent studies have provided indisputable evidence for a role of miRNAs in normal haematopoiesis adding a further layer of complexity to the regulatory process. Leukaemia and lymphoma are characterised by dysregulation of survival and differentiation in haematopoietic progenitor cells. There are several lines of evidence supporting the notion that miRNA dysfunction is contributory, whether by extrapolation from miRNA-mediated oncogenesis in other disorders, by miRNA profiling, or by in vivo and in vitro functional studies of miRNAs in haematological malignancies. Further work is underway to delineate the role of miRNAs in the pathogenesis of malignant blood disorders, with the eventual hope that this will aid in the diagnosis and the development of potential novel cures for such diseases, many of which still have an unacceptable prognosis. PMID- 19291532 TI - Staging bone marrow in diffuse large B-cell lymphoma: the role of ancillary investigations. AB - Diffuse large B-cell lymphoma (DLBCL) is an aggressive non-Hodgkin's lymphoma (NHL) that requires treatment at initial diagnosis. Treatment decisions may be guided by staging investigations, which include assessment of bone marrow (BM). Traditionally this assessment has included light microscopic examination of the bone marrow aspirate and trephine biopsy. Ancillary testing--including immunophenotyping using flow cytometry on BM aspirates, immunohistochemistry (IHC) on BM trephines and molecular studies using immunoglobulin heavy (IgH) or light (IgL) chain gene rearrangements--are performed as required to aid histological diagnosis. This review examines the evidence for the routine use of ancillary tests in the initial staging of DLBCL, including the limitations, potential pitfalls and possible clinical implications of their use in this setting. PMID- 19291533 TI - Primary cutaneous lymphoblastic lymphoma in children: series of eight cases with review of the literature. AB - AIM: Primary cutaneous lymphoblastic lymphoma is a rare but well recognised tumour predominantly of childhood. In this study we examine eight cases of cutaneous lymphoblastic lymphoma in children, which is the largest series to date of tumours confined to the skin with or without local lymph nodes (stage I or II) but without systemic disease at diagnosis. METHODS: The clinical history and histology from the eight cases were reviewed together with a panel of immunohistochemical stains to confirm lineage and diagnosis. RESULTS: Seven of the eight cases were confirmed as B-lymphoblastic lymphoma (B-LBL) of the skin, with the eighth case representing a CD4+/CD56+ plasmacytoid dendritic cell tumour. The cases were all stage I or II, and all patients received systemic chemotherapy after full staging investigations to exclude systemic disease at diagnosis. All patients remained in complete remission at the time of last follow up of between 3 and 9 years from diagnosis. CONCLUSIONS: Lymphoblastic lymphoma may present primarily in the skin without systemic manifestation, with the majority of such cases representing B-LBL. A rare case of childhood CD4+/CD56+ plasmacytoid dendritic cell tumour with similar blastic morphology is also described. Full staging investigations are mandatory at diagnosis to exclude systemic disease. Cases confined to stage I or II at diagnosis carry an excellent prognosis with appropriate systemic chemotherapeutic treatment. PMID- 19291534 TI - Mac-2 binding protein and galectin-3 expression in mucinous tumours of the ovary: an annealing control primer system and immunohistochemical study. AB - AIM: We used a new differential display method, the annealing control primer (ACP) system, to analyse the differentially expressed genes in mucinous ovarian tumours. To verify the corresponding target gene, immunohistochemical staining was performed on various epithelial tumours of the ovary. METHODS AND RESULTS: The ACP-based reverse transcriptase-polymerase chain reaction revealed that 21 genes were upregulated in the mucinous ovarian adenoma and 14 genes were upregulated in the mucinous ovarian carcinoma. Among them, we selected one upregulated gene, the Mac-2 binding protein (Mac-2 BP), and verified the expression of the Mac-2 BP and its ligand, galectin-3, in a variety of epithelial ovarian tumours by immunohistochemistry. Positive expression of the Mac-2 BP was significantly higher in the mucinous ovarian tumours compared to the other epithelial tumours. Mac-2 BP expression was significantly increased in the borderline and malignant tumours compared to the benign tumours. Galectin-3 expression was more frequent in clear cell carcinomas, serous tumours and mucinous tumours than in endometrioid and transitional tumours. However, there were no differences in galectin-3 expression in comparisons among benign, borderline and malignant mucinous and serous tumours. CONCLUSION: These data indicate that the Mac-2 BP may play a role in the development and progression of mucinous ovarian tumours. PMID- 19291535 TI - Pulmonary metastases from uterine neoplasms after long tumour-free interval: four cases and review of the literature. AB - AIMS: Uncommon cases of lung metastasis from different types of uterine neoplasms with a long tumour-free interval after hysterectomy are reported. METHODS AND RESULTS: Four cases were retrieved from our surgical pathology archives. Case 1 was a 68-year-old woman who had three pulmonary nodules 23 years after hysterectomy for low-grade endometrial stromal sarcoma (LGESS). The nodules obtained with video-assisted thoracic surgeries were consistent with metastatic LGESS. Case 2 was a 36-year-old woman who had numerous bilateral pulmonary nodules 6 years after hysterectomy for leiomyoma. A transthoracic biopsy revealed benign metastasising leiomyoma. Case 3 was a 77-year-old woman who had a large lung mass with satellite nodules 17 years after hysterectomy with bilateral salpingo-oophorectomy and subsequent radiotherapy for endometrial endometrioid adenocarcinoma (EEA). The biopsied and resected lung tumour was consistent with metastatic EEA. Case 4 was a 51-year-old woman who underwent total hysterectomy and subsequent radiotherapy for endocervical adenocarcinoma 12 years ago and lung lobectomy for metastatic disease 8 years ago. She then developed two pulmonary lesions 14 months ago, and these resected after radiotherapy were metastatic endocervical adenocarcinoma. CONCLUSIONS: A review of the literature revealed that late pulmonary metastasis from uterine neoplasms is rare but not negligible. Immunohistochemical studies and molecular tests, together with detailed clinical information and imaging findings, are important for rendering a diagnosis. PMID- 19291536 TI - Human papillomavirus is detected in transitional cell carcinoma arising in renal transplant recipients. AB - AIMS: We investigated the role of human papillomavirus (HPV) in the development of transitional cell carcinoma (TCC) arising in renal transplant recipients. METHODS: Genomic DNA was extracted from 10 microm paraffin embedded sections of five TCCs arising in five renal transplant recipients using the QIAamp DNA mini kit according to the manufacturer's instructions. beta-globin PCR was performed to test DNA adequacy. Samples were tested for the presence of HPV DNA by broad spectrum HPV PCR method using non-biotinylated SPF10 primers (SPF1A, SPF1B, SPF1C, SPF1D, SPF2B, SPF2D) which amplify a short 65 bp fragment. Positive bands were identified on a 3% gel. Positive samples underwent a second HPV PCR and were amplified using biotinylated SPF10 primer set, which amplifies the same 65 bp region of the L1 open reading frame. INNO-LiPA line probe assay was then performed to genotype the samples which uses a reverse hybridisation principle. RESULTS: Four of five TCCs examined were positive for HPV. The high risk HPV16 was detected in three cases whereas in the fourth case an unclassifiable HPV genotype was present. In all DNA samples, beta-globin amplification was successful. CONCLUSIONS: Our results indicate that HPV and in particular HPV16 may play an aetiological role in the development of TCC in renal transplant patients. PMID- 19291537 TI - LYVE-1 immunocytochemical staining for gastrointestinal carcinoids. AB - AIMS: By immunocytochemical staining for lymphatic vessels using anti-lymphatic vessel hyaluronan receptor (LYVE-1) antibody, pancreatic islets and some pancreatic endocrine tumours (PETs) were positively stained for the cytoplasm in addition to lymphatic vessels. The current study was extended to investigate possible immunostaining of gastrointestinal carcinoids using goat antihuman LYVE 1 antibody. METHODS: Lymphatic vessels were immunostained by LYVE-1 and blood vessels were immunostained by factor 8 (F-8). RESULTS: Among 27 gastrointestinal carcinoids, six cases (22%) including five primary and one metastatic carcinoids were positive in the cytoplasm for LYVE-1 in addition to lymphatic vessels and there were abundant lymphatic and blood vessels at the margin of the carcinoids. These six LYVE-1 positive cases consisted of three of 10 small intestinal, one of five appendiceal and two of seven large intestinal carcinoids. There was no LYVE 1 positive staining in the adjacent normal gastro-intestinal mucosa. CONCLUSIONS: The presence of LYVE-1 immunostaining in some gastrointestinal carcinoids may support a structure-function relationship of lymphatic vessels/hyaluronan receptor for modulating synthesis and secretion of hormones and amines by carcinoid tumour cells. PMID- 19291538 TI - Diphenylhydantoin plays a role in gene expression related to cytoskeleton and protein adhesion in human normal palate fibroblasts. AB - AIMS: Morphogenetic processes during palate development are related to extracellular matrix composition. The cell-extracellular matrix relation plays a role in cell activity and in gene expression. We studied the effect of diphenylhydantoin, a teratogen known to induce cleft palate in human newborns, on extracellular matrix production. We investigated whether diphenylhydantoin treatment caused any differences in glycosaminoglycans, collagen synthesis and gene expression in human normal palate fibroblasts. METHODS: Human palate fibroblasts were maintained for 24 hours in serum-free 199 medium containing 5 microg/mL (3)H-glucosamine or (3)H proline hydrochloride. Collagen and glycosaminoglycan classes were then measured using biochemical methods, gene expression with microarray analysis and cytoskeleton components with immunofluorescent antibodies and computer analysis. RESULTS: In normal fibroblasts diphenylhydantoin reduced collagen and glycosaminoglycan synthesis with a marked effect on sulphated glycosaminoglycans. There were also substantial decreases in tubulin, vimentin and alpha-actin staining and an increase of vinculin compared to controls. Diphenylhydantoin acted on several genes related to the synthesis of cytoskeleton and adhesion membrane proteins. It inhibited caderin, caveolin, RTK and alpha-actin, and increased nectin, cytoplasmatic FRG vinculin, ITGA, ITGB extracellular matrix ligand and EDG2 gene expression. DNA binding gene expression, which plays a role in cell growth and senescence, was activated. CONCLUSIONS: Since cell activity is dependent on the cell morphology and extracellular matrix composition, these findings indicate that in human normal palate fibroblasts diphenylhydantoin can modify cytoskeletal components and extracellular matrix-cell adhesion, with consequent effects on gene expression. These changes might be related to anomalous palate development. PMID- 19291539 TI - Early renal failure detection by cystatin C in Type 2 diabetes mellitus: varying patterns of renal analyte expression. AB - AIM: The early stages of renal failure are poorly diagnosed by current routine tests. We studied cystatin C and routine renal analyte patterns in Type 2 diabetes mellitus. METHODS: Type 2 diabetes mellitus patients (n = 48) were tested for serum cystatin C, urine albumin, haemoglobin A1c, serum creatinine, serum urea, urine creatinine, glucose, triglycerides and low density lipoproteins (LDL). Glomerular filtration rate (GFR) estimates were made using Cockroft-Gault and Modification of Diet in Renal Disease formulae. RESULTS: The cystatin C (95%CI) reference range was 0.78-0.86 mg/L. While serum cystatin C showed general correlation with routine renal tests, a plateau was observed in analytes measured against cystatin C. Cystatin C improved sensitivity led to detection of renal abnormality in 19% of patients not diagnosed by routine tests. CONCLUSIONS: Cystatin C is a more sensitive marker of renal disease in Type 2 diabetes mellitus where estimated GFR is unreported at >60 mL/min and where antihypertensive medications render microalbuminuria detection unreliable. Its incorporation into a panel of renal function tests is highly recommended. PMID- 19291540 TI - Heat shock protein induction in fetal mouse brain as a measure of stress after whole of gestation exposure to mobile telephony radiofrequency fields. AB - AIM: To determine whether whole of gestation exposure of fetal mouse brain to mobile telephone radiofrequency fields produces a stress response detectable by induction of heat shock proteins (HSPs). METHODS: Using a purpose-designed exposure system at 900 MHz, pregnant mice were given a single, far-field, whole body exposure at a specific absorption rate of 4 W/kg for 60 min/day from day 1 to day 19 of gestation. Control mice were sham-exposed or freely mobile in a cage to control for any stress caused by restraint in the exposure module. Immediately prior to parturition on day 19, fetal brains were collected, fixed in 4% paraformaldehyde and paraffin-embedded. Three coronal sections encompassing a wide range of anatomical regions were cut from each brain and any stress response detected by immunostaining for HSP25, 32 and 70. RESULTS: There was no induction of HSP32 or 70 in any brains, while HSP25 expression was limited to two brainstem nuclei and occurred consistently in exposed and non-exposed brains. CONCLUSION: Whole of gestation exposure of fetal mouse brains to mobile phone radiofrequency fields did not produce any stress response using HSPs as an immunohistochemical marker. PMID- 19291541 TI - Molecular confirmation of pathological specimen integrity in Australasia. AB - AIM: Investigations into 14 suspected pathology sample identification errors and mix-ups were performed, as a service for several public hospital and private laboratories, from 2005 to 2007. METHODS: Analyses were performed with the forensic ABI Identifiler kit of 16 microsatellites (15 autosomal and amelogenin) on DNA from paraffin-embedded tissues or blood specimens and compared to independently verified (single or multiple) patient samples. RESULTS: Of 23 unique patient specimens referred for sample integrity confirmation because of pathologist, clinician or patient concern, six (26.1%) were demonstrated to be discordant, indicating that specimen identification errors or mix-ups had occurred. CONCLUSIONS: Due to their great sensitivity and high discrimination power, forensic identity multiplex systems using either microsatellites or single nucleotide polymorphisms (SNPs) can resolve concerns about pathology specimen identity and integrity. PMID- 19291542 TI - The co-location of an ovarian clear cell carcinoma and mucinous cystadenoma. PMID- 19291543 TI - Synchronous occurrence of primary adenocarcinoma and schwannoma in the stomach: a case report. PMID- 19291544 TI - Metastatic fibroblastic reticulum cell sarcoma of the liver: pathological and PET CT evaluation. PMID- 19291545 TI - Ameloblastic carcinoma (spindle cell variant). PMID- 19291546 TI - Leprosy of the soft palate masquerading as squamous cell carcinoma. PMID- 19291547 TI - A case of hyperIgG4 disease or IgG4-related sclerosing disease presenting as retroperitoneal fibrosis, chronic sclerosing sialadenitis and mediastinal lymphadenopathy. PMID- 19291548 TI - Acute light chain tubulopathy in myeloma. PMID- 19291549 TI - Presacral extramedullary haematopoiesis. PMID- 19291550 TI - Predictive value of the Ontario prehospital stroke screening tool for the identification of patients with acute stroke. AB - BACKGROUND: In 2005, a prehospital stroke screening tool was implemented in Toronto, Ontario, Canada. Patients identified by paramedics through the use of this tool in the field were transported to a regional stroke center under an acute stroke protocol. OBJECTIVE: To determine the positive predictive value (PPV) of the Ontario Prehospital Stroke Screening Tool for identification of acute stroke at a single stroke center. METHODS: We conducted a retrospective analysis of consecutive patients transported to a regional stroke center under the prehospital acute stroke protocol over a 12-month period. Final diagnoses, treatments, and outcomes were abstracted from a provincial registry. Rates of fibrinolysis were compared with those for the 12-month period prior to implementation of the stroke protocol. RESULTS: Three hundred twenty-five patients were triaged under the emergency medical services (EMS) acute stroke protocol over the study period. The PPV of the screening tool was 89.5% (95% confidence interval [CI]: 85.7-92.7%) for acute stroke. Thirty-four patients (11%) had nonstroke conditions, with the most common being seizure (4%). The rate of administration of tissue plasminogen activator (tPA) for all patients with suspected stroke increased from 5.9% to 10.1% (p = 0.04) compared with the rate in the 12-month period prior to implementation of the acute stroke protocol. The tPA rate for patients arriving under the stroke protocol was 17.2%. Most patients (75%) receiving tPA arrived from outside the hospital catchment area. CONCLUSIONS: In this preliminary study, the Ontario Prehospital Stroke Screening Tool had a high PPV for acute stroke and appeared to be effective for identifying patients who required triage to a single regional stroke center. Following implementation of a citywide acute stroke protocol using this screening tool, we observed an increase in the number of patients who were eligible for and received fibrinolysis at our stroke center. PMID- 19291551 TI - Emergency medical service providers' attitudes and experiences regarding enrolling patients in clinical research trials. AB - OBJECTIVE: The purpose of this study was to evaluate Emergency Medical Services (EMS) providers' attitudes and experiences about enrolling patients in clinical research trials utilizing the federal rules for exception from informed consent. We hypothesized that Emergency Medical Technicians (EMTs) would have varied attitudes about research using an exception from informed consent which could have an impact on the research. METHODS AND SETTING: Since January 2007, the EMS system has been participating in a randomized, multi-center interventional trial in which out-of-hospital providers enroll critically injured trauma patients using exception from informed consent.A voluntary, anonymous, written survey was administered to EMS providers during an in-service. The survey included demographics and Likert-type questions about their experiences with and attitudes towards research in general, and research using an exception from informed consent for an out-of-hospital clinical trial. RESULTS: The response rate was 79.3% (844/1067). Most respondents, 93.3%, agreed that "research in EMS care is important." However, 38.5% also agreed that individual EMTs/paramedics should maintain the personal right of refusal to enroll patients in EMS trials. Fifty four percent of respondents agreed with the statement that "the right of research subjects to make their own choices is more important than the interests of the general community." In response to statements about the current study, 11.3% agreed that "the study is unethical because the patient cannot consent" and 69.2% responded that they would personally be willing to be enrolled in the study before they were able to give consent if they were seriously injured. Those who had not enrolled a patient into the study (681 respondents) were asked their reasons: 76.8% had not encountered an eligible patient or did not work for an agency that carried the fluid; 4.3% did not have time; 4.1% forgot and 1.1% stated that they were opposed to enrolling patients in studies without their consent. CONCLUSION: The majority of EMS personnel in one community support EMS research and this specific out-of-hospital clinical trial being conducted under an exception from informed consent. Potential barriers to enrollment were identified. Further study in other systems is warranted to better understand EMS provider perspectives about exception from informed consent research. PMID- 19291552 TI - Potential impact of a verbal prehospital DNR policy. AB - BACKGROUND: Forgoing resuscitation in prehospital cardiac arrest has previously required a written prehospital do-not-resuscitate (DNR) order. Some emergency medical services (EMS) agencies, including Los Angeles County (LAC), have implemented policies allowing surrogate decision makers to verbally request to forgo resuscitation. The impact of a verbal DNR policy is unclear, given the absence of information about how often cardiac arrest occurs at home, or in the presence of a family member. OBJECTIVE: To determine the prevalence of written DNR forms, rate of resuscitation, location of cardiac arrest, and availability of a family member in nontraumatic cardiac arrest prior to implementation of the new policy in LAC. METHODS: All prehospital run sheets for nontraumatic cardiac arrest in LAC were reviewed for the first seven days of each month (August 2006 January 2007) for DNR status, location of cardiac arrest, presence of family members, and whether resuscitation was attempted. RESULTS: Of the 897 cardiac arrests, 492 occurred at home, 111 in a public place, and 93 in a nursing home (location was unknown for 201). Fifty-five patients (6%) had a written DNR order, although it was not always available. Of these 55 patients, ten were resuscitated, the majority of the time because the family could not produce the paperwork. A family member was listed as present 29% of the time (261 of 897 cases). A medical history was obtained in an additional 465 cases (52%), indicating that someone familiar with the patient's medical history was present more than half the time, even when a family member was not mentioned. CONCLUSIONS: A written DNR order is uncommonly used in the prehospital setting as a reason to forgo resuscitation in LAC. Even when family members state that the patient has a DNR order, patients are often resuscitated. A majority of cardiac arrests occurs at the patient's home, and in many cases in the presence of family members, some of whom may be able to express a patient's preferences regarding end-of-life care. PMID- 19291553 TI - Repeat emergency medical services use by older adults in a rural community: impact on research methods and study length. AB - OBJECTIVES: To evaluate the proportion of emergency medical services (EMS) requests in a rural community made by unique, noninstitutionalized older adults, or individuals making their first request for EMS assistance during the study period, and the impact on research parameters. METHODS: This study was a retrospective chart review of patients aged 65 years and older cared for by the Geneseo Fire Department Ambulance between February 2004 and May 2005 (period 1) and between July 2006 and October 2007 (period 2). The Geneseo Fire Department Ambulance response territory is a rural community in Upstate New York. We obtained demographic information including age, race, gender, call location, and the frequency of EMS use from the medical record, as well as clinical information including level of prehospital care, chief complaint, and disposition. Descriptive statistics were used for the analysis, along with 95% confidence intervals. RESULTS: Over two 16-month periods, approximately 70% of the EMS calls by community-dwelling (noninstitutionalized) older adults were from unique individuals. The monthly proportion ranged from 75-100% during the first four months to 43-80% for the remaining 12 months for both groups. CONCLUSION: In rural, prehospital studies that enroll older adults and last more than four months, approximately 70% of EMS requests are made by unique older adults, or individuals making their first request for EMS assistance. Investigators must consider these results when estimating the enrollment period for prehospital studies. PMID- 19291554 TI - Management of prehospital seizure patients by paramedics. AB - BACKGROUND: Seizure patients are frequently encountered in the prehospital environment and have the potential to need advanced interventions, though the utility of advanced life support (ALS) interventions in many of these patients has not been proven. OBJECTIVE: Our goals were to assess the management of prehospital seizure patients by paramedics in an urban EMS system with an existing ALS-based prehospital seizure protocol and to assess characteristics and short-term outcomes that may aid in addressing the utility of specific ALS interventions. METHODS: This was a retrospective study of 97 EMS cases with the chief complaint of seizure. Prehospital records were reviewed for patient and event characteristics, including past seizure history, seizure timing, level of consciousness, on-scene and transport times, and EMS interventions. Emergency department (ED) records were reviewed for recurrence of seizure activity, ED evaluation, and disposition. Data were analyzed using descriptive statistics and Student t-test. RESULTS: Of 87 patients meeting the protocol inclusion criteria for all ALS interventions, 11 (12.6%) received cardiac monitoring, 55 (63.2%) had intravenous (IV) access attempted, and 56 (64.4%) had blood glucose determination. Average on-scene time was 5.9 minutes longer if IV access was attempted (p = 0.001), though transport times were not significantly different (11.6 versus 11.3 minutes, respectively; p = 0.851). Additional seizure activity occurred in the prehospital and/or ED settings in 28 patients (28.9% of all cases), including 17 in the prehospital setting and 15 in the ED. Diazepam was administered by EMS for half of the eight (8.2%) patients who had seizures lasting more than 1 minute, while the remainder had seizures that were focal or spontaneously resolved. CONCLUSION: This study showed a lower-than-anticipated level of compliance with an ALS-based prehospital seizure protocol, though patient-specific care appeared appropriate. Prehospital seizure patients have the potential for seizure recurrence and may benefit from focused ALS interventions, but their heterogeneity makes uniform protocols difficult to develop and follow. PMID- 19291555 TI - Physician medical direction and clinical performance at an established emergency medical services system. AB - OBJECTIVE: Few developed emergency medical services (EMS) systems operate without dedicated medical direction. We describe the experience of Hamad Medical Corporation (HMC) EMS, which in 2007 first engaged an EMS medical director to develop and implement medical direction and quality assurance programs. We report subsequent changes to system performance over time. METHODS: Over one year, changes to the service's clinical infrastructure were made: Policies were revised, paramedic scopes of practice were adjusted, evidence-based clinical protocols were developed, and skills maintenance and education programs were implemented. Credentialing, physician chart auditing, clinical remediation, and online medical command/hospital notification systems were introduced. RESULTS: Following these interventions, we report associated improvements to key indicators: Chart reviews revealed significant improvements in clinical quality. A comparison of pre- and post-intervention audited charts reveals a decrease in cases requiring remediation (11% to 5%, odds ratio [OR] 0.43 [95% confidence interval (CI) 0.20-0.85], p = 0.01). The proportion of charts rated as clinically acceptable rose from 48% to 84% (OR 6 [95% CI 3.9-9.1], p < 0.001). The proportion of misplaced endotracheal tubes fell (3.8% baseline to 0.6%, OR 0.16 [95% CI 0.004-1.06], (exact) p = 0.05), corresponding to improved adherence to an airway placement policy mandating use of airway confirmation devices and securing devices (0.7% compliance to 98%, OR 714 [95% CI 64-29,334], (exact) p < 0.001). Intravenous catheter insertion in unstable cases increased from 67% of cases to 92% (OR 1.31 [95% CI 1.09-1.71], p = 0.004). EMS administration of aspirin to patients with suspected ischemic chest pain improved from 2% to 77% (OR 178 [95% CI 35-1,604], p < 0.001). CONCLUSIONS: We suggest that implementation of a physician medical direction is associated with improved clinical indicators and overall quality of care at an established EMS system. PMID- 19291556 TI - Bispectral index monitoring in helicopter emergency medical services patients. AB - Background. Many critically ill patients are given sedatives and paralytics to facilitate aeromedical transport. Bispectral index (BIS) monitoring is a computer derived electroencephalography (EEG) analog currently used to monitor the level of awareness of sedated patients. It gives a score of 1-100, with 1 representing no brain function and 100 representing a completely alert patient. Objective. To evaluate whether critically ill patients are adequately sedated during aeromedical transport. Methods. This was a prospective, observational study of a convenience sample of critically ill patients transported by helicopter. All intubated patients who received sedatives and/or paralytics to facilitate transport were eligible for enrollment by the attending clinician. Prior to liftoff, a BIS sensor was applied to the patient's forehead. Minimum, maximum, and mean BIS index scores were recorded every minute during transport. Results. Forty-seven patients (57% male) were enrolled, with a median age of 60 years (interquartile range [IQR] 18-81, range 14 to 86 years). The median duration of monitoring was 15.0- minutes (IQR 6.0-26.0, range 2 to 33). The median BIS score was 54.6 (IQR 38.6-67.3, range 28 to 89.5). Only two patients (4.3%, 95% confidence interval [CI] 0.5% to 14.8%) had at least one BIS score greater than 85, the accepted threshold for recall. Conclusion. These results suggest that patients are adequately sedated during air medical transport. PMID- 19291557 TI - Noninvasive ventilation during pediatric interhospital ground transport. AB - OBJECTIVE: We report our use of noninvasive ventilation (NIV) during pediatric interhospital ground transport. METHODS: We retrospectively reviewed transport and hospital records for nonneonatal patients 0.05). CONCLUSIONS: Paramedics in an urban/suburban EMS system can diagnose STEMI and identify appropriate cardiac catheterization laboratory activations with a high degree of accuracy, and an acceptable false-positive rate, when tested using paper-based scenarios. PMID- 19291560 TI - Implementation of specialty centers for patients with ST-segment elevation myocardial infarction. AB - BACKGROUND: Early percutaneous coronary intervention (PCI) has been shown to be superior to fibrinolytic therapy and is associated with reduced morbidity and mortality for patients with ST-segment elevation myocardial infarction (STEMI). OBJECTIVE: To determine the performance of a regional system with prehospital 12 lead electrocardiogram (ECG) identification of STEMI patients and direct paramedic transport to STEMI receiving centers (SRCs) for provision of primary PCI. METHODS: This was a prospective study evaluating the first year of implementation of a regional SRC network to determine the key time intervals for patients identified with STEMI in the prehospital setting. Results. During the 12 month study period, 1,220 patients with a suspected STEMI were identified on prehospital 12-lead ECG, of whom 734 (60%) underwent emergency PCI. A door-to balloon time of 90 minutes or less was achieved for 651 (89%) patients, and 459 (62.5%) had EMS-patient contact-to-balloon times or = 30) and 90 age matched patients with normal weight (BMI 19 - 26) who underwent heart transplantation at our institution between January 1997 and December 2005. RESULTS: Morbidly obese patients experienced higher rates of pre-transplant diabetes (29% vs 15%, p < 0.05) and prolonged waiting time before transplantation (191.4+/-136.1 vs 117.4+/ 143.2 days, p < 0.001). There were no significant differences in post-operative complications including rejection and major and minor infections. There was no difference in actuarial survival between the obese and control groups after a mean follow-up of 4.26+/-2.95 years (p = 0.513, log-rank statistic 0.452). Causes of death did not differ. Cox proportional hazard analysis revealed increased association of peripheral vascular disease (HR 31.718, p = 0.001), and pre operative inotropic support (HR 33.725, p = 0.013) with increased mortality in the obese group. CONCLUSIONS: This study suggests morbid obesity does not affect survival or rates of infection and rejection after heart transplantation. PMID- 19291587 TI - Laboratory surveillance of urogenital Chlamydia trachomatis infections in Denmark 1988-2007. AB - In 1994 laboratory reporting of urogenital chlamydia was integrated in the countrywide notification system for infectious diseases. Previously (1988-1993), laboratory confirmed cases had been reported on a voluntary basis. The applied laboratory technologies changed significantly: in 1988, 48% of cases were diagnosed by culture; in 1994, 88% by enzyme immunoassays; and since 2004 more than 99% by nucleic acid amplification tests. The proportions of chlamydia positive men diagnosed by testing urine were <1% in 1994, 10% in 2001 and 75% in 2007. From 1994 to 2002 the annual incidence rates of chlamydia per 100,000 population among women were around 350-400 and then increased to 586 in 2007. Among men the incidence rate rose from 125 in 1994 to 358 in 2007. In conclusion, the annual incidence rates of urogenital chlamydia remained high and possibly increasing during a 14-y period (1994-2007), with a high testing activity, improving technologies and countrywide information campaigns. The observed increase is likely to be associated with the introduction of more sensitive diagnostic methods and with increasing testing of men. A possible change in target groups from low- to high-prevalence could not be assessed due to lack of information about persons being tested. PMID- 19291588 TI - A new life with aphasia: everyday activities and social support. AB - Abstract People who develop aphasia must adjust their lifestyles and learn to cope with the activity limitations that may follow from their disability. The purpose of this study was to describe aphasic individuals' experiences of everyday activities and social support in daily life. Interviews were conducted with 20 people with aphasia, and analysed with qualitative content analysis. The results show that everyday activities changed considerably with the onset of aphasia, and the participants were hindered from participating in activities by communication problems or physical disabilities. Aphasia led to the loss of friends and colleagues, and the interviewees often felt lonely. They generally received a lot of social support from close relatives, but support from the healthcare system was lacking. They need different kinds of social support to help them manage their aphasia and everyday activities and to improve their participation in society. Further studies are needed to improve our knowledge of everyday activity and social support for people with aphasia, and what it means to live with aphasia. PMID- 19291589 TI - Prescribing behaviour after the introduction of decentralized drug budgets: is there an association with employer and type of care facility? AB - OBJECTIVE: To analyse whether prescribing patterns changed after introduction of drug budgets and whether there is an association between drug prescribing patterns and the type of employer and care facility. METHODS: Data analysed encompassed information on dispensed medicines, by workplaces, prescribed in the Region Vastra Gotaland, Sweden, for the years 2003 and 2006. Workplaces (n = 969) were categorized according to type of employer and type of care facility. Five prescribing indicators reflecting goals for cost-containing prescribing in Region Vastra Gotaland were assessed. Changes over time and differences between different types of employer and care facility were analysed by Mann-Whitney tests. RESULTS: In 2003, workplaces with a public employer had a significantly higher adherence to three of the prescribing indicators compared with private practitioners. Two of these differences remained in 2006. In 2003, none of the prescribing indicators differed between primary care and other care facilities. Three years later workplaces in primary care had a significantly higher adherence to three of the prescribing indicators than other care facilities. There was a statistically significant difference in change between 2003 and 2006 between primary care and other care facilities; there were no differences in change between workplaces with public and private employers. CONCLUSIONS: Adherence to three of the prescribing indicators increased after the introduction of decentralized drug budgets. Workplaces with a public employer showed greater adherence to two of the prescribing indicators than private sector workplaces. PMID- 19291590 TI - Validation of a new plasma cystatin C-based formula and the Modification of Diet in Renal Disease creatinine-based formula for determination of glomerular filtration rate. AB - OBJECTIVE: New proposed definitions of chronic kidney disease necessitate the development and use of simple and accurate methods for estimating glomerular filtration rate (GFR). Plasma cystatin C has been shown to be a more reliable GFR marker than creatinine and formulae for estimating GFR have been reported. The purpose of this study was to validate a cystatin C-based GFR prediction equation in a different population from the derivation set but using the cystatin C assay of a single laboratory, and to compare the results with that of the creatinine based Modification of Diet in Renal Disease (MDRD) Study equation. MATERIAL AND METHODS: A newly presented formula based on plasma cystatin C and gender and the MDRD formula based on creatinine for estimation of GFR were validated in an unselective patient material. Single sample iohexol clearance was used as the GFR reference method in 406 consecutive patients with GFR varying from normal to poor renal function. The creatinine assay used was standardized to express true plasma creatinine. RESULTS: Median bias (1.1%) and accuracy (79.1% of the estimates within +/-30% of iohexol clearance) of the cystatin C formula were close to the derivation set. The accuracy was significantly higher than that of the original four-variable MDRD equation (73.2%; median bias 9.8%). However, the accuracy did not differ significantly from that of the re-expressed MDRD formula (79.6%; median bias 3.2%) based on true creatinine. Both formulae performed with a low bias and acceptable accuracy up to a GFR of 90 ml/min/1.73 m(2). CONCLUSIONS: GFR estimation based on plasma cystatin C performed equally well in the validation as in the derivation set, and was as accurate as the re-expressed MDRD creatinine based equation. PMID- 19291591 TI - Dopamine (DA) induced irreversible proteasome inhibition via DA derived quinones. AB - This study demonstrated that DA and its oxidative metabolites: H2O2 and aminochrome (AM), cyclized DA quinones, could all directly inhibit proteasome activity. DA and AM, especially AM, could induce intensive and irreversible proteasome inhibition, whereas proteasome inhibition induced by H2O2 was weaker and GSH reversible. It was concluded that DA induced irreversible proteasome inhibition via DA-derived quinones, rather than through small molecular weight ROS. The AM was also more toxic than H2O2 to dopaminergic MN9D cells. Furthermore the cytotoxicity and proteasome inhibition induced by DA, AM and H2O2 could be abrogated by GSH, ascorbic acid (AA), Vitamin E, SOD (superoxidase dismutase) or CAT (catalase) with different profiles. Only GSH was potent to abrogate DA, AM or H2O2-induced cell toxicity and proteasome inhibition, as well as to reverse H2O2 induced proteosome inhibition. Therefore, therapeutic strategies to increase GSH level or to use GSH substitutes should function to control PD onset and development. PMID- 19291592 TI - Glutathionylation regulates cytosolic NADP+-dependent isocitrate dehydrogenase activity. AB - Cytosolic NADP+-dependent isocitrate dehydrogenase (IDPc) is susceptible to inactivation by numerous thiol-modifying reagents. This study now reports that Cys269 of IDPc is a target for S-glutathionylation and that this modification is reversed by dithiothreitol as well as enzymatically by cytosolic glutaredoxin in the presence of GSH. Glutathionylated IDPc was significantly less susceptible than native protein to peptide fragmentation by reactive oxygen species and proteolytic digestion. Glutathionylation may play a protective role in the degradation of protein through the structural alterations of IDPc. HEK293 cells treated with diamide displayed decreased IDPc activity and accumulated glutathionylated enzyme. Using immunoprecipitation with an anti-IDPc IgG and immunoblotting with an anti-GSH IgG, we purified and positively identified glutathionylated IDPc from the kidneys of mice subjected to ischemia/reperfusion injury and from the livers of ethanol-administered rats. These results suggest that IDPc activity is modulated through enzymatic glutathionylation and deglutathionylation during oxidative stress. PMID- 19291593 TI - Neuroprotective potential of combination of resveratrol and 4-amino 1,8 naphthalimide in experimental diabetic neuropathy: focus on functional, sensorimotor and biochemical changes. AB - The present study investigated whether combination of resveratrol and 4-amino 1,8 naphthalimide (4-ANI) is effective in the development of diabetic neuropathy (DN). After 6 weeks of diabetes induction, rats were treated for 2 weeks with resveratrol and 4-amino 1,8 naphthalimide (4-ANI) either alone or in combination. Experimental end points included functional, behavioural and biochemical parameters along with PAR immunohistochemistry and were performed at the end of treatment. Combination of resveratrol (10 mg/kg) and 4-ANI (3 mg/kg) attenuated conduction and nerve blood flow deficits and resulted in amelioration of diabetic neuropathic pain. Significant reversal of biochemical alterations (peroxynitrite, MDA and NAD levels) were also observed, as well as PAR accumulation in the sciatic nerve. This study suggests the beneficial effect of combining resveratrol and 4-ANI in experimental diabetic neuropathy. PMID- 19291595 TI - Isolation and clinical significance of Pseudallescheria and Scedosporium species. AB - In the course of the past decades, Pseudallescheria/Scedosporium species have become increasingly recognized as causative agents of significant infections in humans. In our laboratory in Bonn, Germany, the first clinical strain was isolated by chance by dipping a wooden swab taken from a chronic wound process into brain-heart infusion broth. The latter was being used as an enrichment medium for the recovery of infectious agents. Subsequently, a total of 35 additional Pseudallescheria/Scedosporium strains from four patients suffering from invasive fungal infection were isolated with this method. In contrast, only 27 of the 36 isolates were detected by plating the specimens on Sabouraud glucose agar (75%). Furthermore, eight strains were recovered from the respiratory tract samples of six out of 42 patients suffering from cystic fibrosis. While all eight strains were isolated on SceSel+ agar, only five were obtained from wooden sticks dipped into broth for enrichment of infectious agents (62.5%) and only three on Sabouraud glucose agar (37.5%). The clinical cases are summarized and methods used for detection are described. Species identification is based on the taxonomy valid in 2005; in the interim several re-classifications have been proposed. PMID- 19291596 TI - Using aCGH to study intraspecific genetic variability in two pathogenic molds, Aspergillus fumigatus and Aspergillus flavus. AB - We have examined the feasibility of using array comparative genomic hybridization (aCGH) to explore intraspecific genetic variability at the genomic level in two pathogenic molds, Aspergillus fumigatus and Aspergillus flavus. Our analysis showed that strain-specific genes may comprise up to 2% of their genomes in comparison to isolates from different vegetative (heterokaryon) compatibility groups (VCGs). In contrast, isolates with the same VCG affiliations have almost identical gene content. Most isolate-specific genes are annotated as 'hypothetical' and located in a few large subtelomeric indels. The list includes highly polymorphic loci that contain putative het (heterokaryon compatibility) loci, which determine the individual's VCG during parasexual crossing. Incidentally, VCGs in both species seem to be significantly associated with either alpha or HMG mating type (Chi-square test, P=0.05). In conclusion CGH can be used to effectively to identify isolate-specific genes in Aspergillus species. Preliminary evidence suggests that gene flow in both species is largely constrained by VCG boundaries, although further VCG sampling is required to confirm this observation. PMID- 19291598 TI - Aspergillus terreus complex. AB - Aspergillus terreus can cause invasive infections in humans, which are often refractory to therapy with the antifungal drug amphotericin B and have a propensity to dissemination. The organism has diverse colony morphology, and sub typing studies have demonstrated that isolates of A. terreus are diverse in their genotypes. However, detailed phylogenetic studies of section Terrei employing sequence information from protein coding regions have not been thus far attempted. Interestingly, A. terreus produces unicellular forms called accessory conidia in vitro and during infection; the clinical relevance of these structures are not well understood. This paper presents an overview of the morphology, species identification strategies, and molecular epidemiology of A. terreus. PMID- 19291600 TI - Ormond's disease or secondary retroperitoneal fibrosis? An overview of retroperitoneal fibrosis. AB - Retroperitoneal fibrosis represents a rare inflammatory disease. About two thirds of all cases seem to be idiopathic (= Ormond's disease). The remaining one third is secondary and may be ascribed to infections, trauma, radiation therapy, malignant diseases, and the use of certain drugs. Up to 15 % of patients have additional fibrotic processes outside the retroperitoneum. The clinical symptoms of retroperitoneal fibrosis are non-specific. In sonography retroperitoneal fibrosis appears as a retroperitoneal hypoechoic mass which can involve the ureters and thus cause hydronephrosis. Intravenous urography and MR urography can demonstrate the typical triad of medial deviation and extrinsic compression of the ureters and hydronephrosis. CT and MRI are the modalities of choice for the diagnosis and follow-up of this disease. The lesion typically begins at the level of the fourth or fifth lumbar vertebra and appears as a plaque, encasing the aorta and the inferior vena cava and often enveloping and medially displacing the ureters. In unenhanced CT, retroperitoneal fibrosis appears as a mass that is isodense with muscle. When using MRI, the mass is hypointense in T 1-weighted images and of variable intensity in T 2-weighted images according to its stage: it may be hyperintense in early stages, while the tissue may have a low signal in late stages. After the administration of contrast media, enhancement is greatest in the early inflammatory phase and minimal in the late fibrotic phase. Dynamic gadolinium enhancement can be useful for assessing disease activity, monitoring response to treatment, and detecting relapse. To differentiate retroperitoneal masses, diffusion-weighted MRI may provide useful information. PMID- 19291601 TI - [Septal delineation using a 20 % diluted contrast chaser bolus in coronary CT angiography: a comparison of 64-slice and dual source CT]. AB - PURPOSE: Due to the technical advance of multislice cardiac CT and the recently introduced dual source CT, the acquisition times for cardiac CT exams are < 10 sec. At the same time the assessment of left ventricular function is possible. However, in many patients a complete right ventricular outwash is noted, leading to insufficient septal delineation. Studies were able to demonstrate that contrast media (CM) mixed in the saline chaser bolus is sufficient for significantly better visualization of septal structures. The aim of this study was to investigate whether this dual flow concept works as well for 64-slice and dual source CT angiography using only 20 % CM in the saline chaser bolus. MATERIALS AND METHODS: 97 patients were included in this prospective study. 47 patients underwent 64-slice CT coronary angiography. 80 cc were administered at 5 cc/sec as the main bolus followed by a 50 cc saline chaser bolus containing 20 % contrast media. The other 50 patients were examined using dual source CT. They received a CM protocol adapted for the scan time with the identical saline/CM chaser bolus. The datasets were quantitatively examined in defined ROIs along the septum and in the right ventricle with respect to the density. The septal delineation was qualitatively analyzed and both groups were compared. In a final step the density was measured in the proximal and distal RCA as well as in the LAD and also compared. RESULTS: Using the protocol adapted for the scan time, significantly less CM was used. No significant difference was able to be found regarding the septal delineation or coronary enhancement. CONCLUSION: The study shows that the dual flow concept allows for robust septal delineation regardless of the CM injection protocol used as long as a 20 % saline chaser bolus is used. A CM protocol adapted for the scan time also leads to significant CM reduction at equal image quality. PMID- 19291602 TI - [Initial experience with prospectively triggered, sequential CT coronary angiography on a 128-slice scanner]. AB - PURPOSE: Spiral CT angiography (CTA) of the coronaries using low-pitch scanning and ECG-gated image reconstruction is a robust method for detecting or excluding relevant coronary plaque. However, the resulting dose exposure is considerable. The aim of the present study was to evaluate image quality and artifacts as well as to record dose values for sequential coronary CTA using a 128-slice scanner with a temporal resolution of 150 ms. MATERIALS AND METHODS: 20 patients with a regular heart rate and without contraindications for oral/I.V. beta blockers, who were referred for CTA of the coronaries for exclusion or detection of relevant plaques, were examined by sequential CTA with the following parameters: 120 kV, 200 ref mAs, collimation 2 x 64 x 0.6, table feed of 34.5 mm at a detector width of 38.4 mm. A total acquisition time of 380 ms per table position allowed for mild shifting of the reconstruction window within the cardiac cycle of +/- 5 %. 50 ml of contrast agent were injected at 5 ml/s followed by a 50 ml split bolus (20 % contrast). The individual start delay was determined by a test bolus scan (10 ml contrast + 50 ml saline flush at 5 ml/s). The image quality for each segment, coronary artery, and patient was determined on a 4-point scale. Dose values were estimated based on the individual dose length product as provided by the scanner's patient protocol. Artifacts were evaluated to determine the cause (calcium vs. motion). RESULTS: All patients received beta blocker pretreatment. The mean heart rate was 62 +/- 5 beats/min. 5 % (13 / 286) of all segments in 5 / 20 patients were rated as non-diagnostic. The mean dose length product was 213 mGy x cm, and the mean effective dose was 3.6 mSv. Calcifications were the major cause of non-diagnostic images. However breathing or other motion artifacts occurred as well. CONCLUSION: In select patients with effective heart rate control and thorough instruction for breath hold compliance, sequential CTA of the coronaries using a 128-slice scanner with a temporal resolution of 150 ms is technically feasible. The resulting effective dose values are clearly below those of spiral coronary CT scans. PMID- 19291603 TI - Strategies for dose-optimized imaging in pediatric cardiac dual source CT. AB - PURPOSE: As a result of rapid advances in CT technology, including dual source CT, cardiac CT exams at high heart rates are feasible, making pediatric cardiac CT in congenital heart disease possible. Dose concerns and patient size variability make general recommendations difficult. The aim of this study was to investigate which scanner settings are suitable for multiple weight groups to provide dose-optimized scanning. MATERIALS AND METHODS: All 12 patients underwent a contrast-enhanced gated dual source CT exam. A maximum of 2 ml/kg body weight or 60 ml contrast was applied. Scanner settings at 80 and 120 kV, as well as weight-adapted mAs settings were used in an iterative fashion. Datasets were assessed for image quality and dose-optimized scan parameters were established via class comparison. In a final step a table was created as a recommendation for cardiac CT in children corresponding to their body weight. Strategies for optimized contrast application with hand vs. manual injection were explored. RESULTS: In all children diagnostic image quality was obtained. In children < 60 kg, 80 kV can be used, in all others 100 kV is sufficient. The eff. mAs varied from 362 mAs to 30 mAs depending on body weight. Retrospectively a significant dose reduction would have been possible in 6 / 12 patients. Cardiac CT can be performed between 0.6 and 3.2 mSv in a pediatric population. CONCLUSION: The present study shows that dual source cardiac CT is a potential diagnostic alternative in children with congenital heart disease without excessive radiation exposure or contrast media application. PMID- 19291604 TI - Iodixanol induces apoptotic and antiproliferative effects but no necrotic cell death in renal proximal tubular cells in vitro. AB - PURPOSE: To evaluate the cytotoxic effects of the iso-osmolar contrast medium iodixanol on renal tubular cell cultures. MATERIALS AND METHODS: LLC-PK1 cells were incubated with iodixanol and isotonic NaCl (18.75 - 75 mg I/ml, 1 - 24 hours). Cell death was assessed by the trypan blue exclusion test. To assess apoptosis, mononucleosomes and oligonucleosomes of cell lysates were determined. Measurement of BrdU (5-bromo-2'-deoxyuridine) incorporation into the DNA was used for the quantification of cell proliferation. RESULTS: Iodixanol did not induce any significant increase in the number of necrotic cells (8 % and 9 % at 37.5 and 75 mg I/ml vs. 8 % for control, p > 0.05). In contrast, iodixanol significantly increased the number of oligonucleosomes indicating induction of apoptosis (125 +/- 4 % of control, p < 0.05). Iodixanol induced a significant, dose- and time dependent inhibition of BrdU incorporation indicating the inhibition of cell proliferation (92 +/- 2 % and 79 +/- 2 % of control at 18.75 and 37.5 mg I/ml, p < 0.001). CONCLUSION: Apoptosis plus an antiproliferative effect of iodixanol without cell necrosis contribute to the renal tubular toxicity of iodixanol. PMID- 19291605 TI - [Dual-source CT: in vitro characterization of gallstones using dual energy analysis]. AB - PURPOSE: Despite clinically available high-resolution CT, the detection and classification of gallstones remains a challenge in some cases. This pilot study examines whether noninvasive characterization of gallstones in vitro is possible using dual-energy analysis (DECT) of dual source CT datasets. MATERIALS AND METHODS: A total of 43 gallstones (0.4 - 1.5 cm) were examined at 80 kV, 140 kV and in the dual-energy mode. The monoenergetic datasets were examined by two independent examiners and classified as calcium, cholesterol or pigment stones. The results were compared with the pathological classification as the clinical gold standard. After creating reference images for each group via dual-energy analysis, the classification was repeated and compared with the gold standard again. RESULTS: Using the monoenergetic analysis at 80 kV, the sensitivity and specificity were 100 / 84 % and 100 / 88 % for calcium stones. For cholesterol stones the values were 54 / 89 % and 54 / 85 % and for pigment stones 70 / 80 % for both examiners. At 140 kV, the sensitivity and specificity for calcium stones were 100 / 84 % for both examiners, 46 / 92 % for cholesterol stones for both examiners and the sensitivity and specificity were 80 / 75 % and 80 / 80 % for pigment stones. Using the reference images established by DECT, both examiners were able to correctly classify all gallstones. CONCLUSION: The present data indicates that DECT is able to correctly classify Gallstones according to the clinical gold standard in vitro. Clinical studies have to demonstrate whether these results lead to optimized clinical decision making. PMID- 19291606 TI - [Asymptomatic siliconoma after prosthesis rupture: portrayal using elastography]. PMID- 19291607 TI - [Multicenter analysis of tolerance and clinical safety of the extracellular MR contrast agent gadobenate dimeglumine (MultiHance)]. AB - PURPOSE: Retrospective analysis of the occurrence of adverse events and the diagnostic efficacy of a paramagnetic contrast agent with weak intermittent protein binding and high relaxivity. MATERIALS AND METHODS: Postmarketing surveillance studies for gadobenate dimeglumine (MultiHance, BRACCO Altana Pharma, Constance) were conducted in Germany between 1998 and 2006 and then retrospectively analyzed. Demographic data, relevant comorbidities, and allergies were recorded. The safety and tolerability of MultiHance were logged on a standardized data sheet. RESULTS: A total of 38,568 patients were included in the study. 829 patients (2.1%) had a known intolerance against contrast media. The examined regions included the central nervous system, the liver, and the vascular bed. The injection rate with automated injectors (n = 10456) varied between 1.0 und 3.0 ml/sec in 86.5% of patients. Adverse events totaled 1.2%. 11 patients (0.03%) experienced serious adverse events. The most frequent findings were nausea, vomiting and a feeling of warmth. CONCLUSION: MultiHance is a safe and very well tolerated contrast agent for magnetic resonance imaging (MRI) with a profile and frequency of adverse events similar to other extracellular MR contrast materials. PMID- 19291608 TI - Cytotoxic activity of isoliquiritigenin towards CCRF-CEM leukemia cells and its effect on DNA damage. AB - The cytotoxic activity of isoliquiritigenin (ISL) towards CCRF-CEM human T-cell leukemia cells and DNA damage induction were investigated in the present study. The anti-proliferative effect of ISL on CCRF-CEM cell line IN VITRO was analyzed. In order to further explore the underlying mechanism of cell growth inhibition of ISL towards CCRF-CEM cell line, the cell cycle distribution and mitochondrial membrane potential (DeltaPsim) disruption were measured. ISL exerted significant activity towards CCRF-CEM cells. The growth inhibitory ratio was concentration- and time-dependent with an IC(50) value of 18.38 microM. The cell cycle was arrested in the G2/M phase upon treatment with low doses of ISL. Mitochondrial membrane potential (DeltaPsim) disruption was observed at low ISL doses. The effect of ISL on DNA damage was analyzed by agarose gel electrophoresis and atomic force microscopy (AFM). Either ISL or Cu2+ failed to cause pBR322 DNA damage. However, plasmid DNA was damaged after treatment with ISL in the presence of Cu2+. Two forms of plasmid DNA closed circular and linear forms were visualized by AFM. PMID- 19291609 TI - 20S-protopanaxadiol inhibits P-glycoprotein in multidrug resistant cancer cells. AB - One of the major causes for cancer cells to resist current chemotherapy is attributed to the over-expression of P-glycoprotein (P-gp), resulting in insufficient drug delivery to the tumor sites. Protopanaxadiol ginsenosides Rg3 and Rh2 are known to induce apoptosis and significantly enhance the tumor inhibitory effects of chemotherapeutics in a synergistic fashion. One of the possible mechanisms is by blocking P-gp activity. The final deglycosylation metabolite of protopanaxadiols (PPDs) IN VIVO is 20S-protopapanaxadiol (aglycone PPD, aPPD), which has also shown anticancer activity and synergy with chemotherapy drugs. In the present study, P-gp over-expressing cancer cells were utilized to test whether aPPD also inhibits P-gp activity. We found that aPPD caused similar cytotoxicity in P388adr cells as their parental non-MDR cells, suggesting that aPPD may not be a substrate of P-gp. On the other hand, the calcein AM efflux assay showed that aPPD was able to inhibit P-gp activity as potently as verapamil on MDR cells. The blockage of P-gp activity was highly reversible as wash-out of aPPD resulted in an immediate recovery of P-gp activity. Unlike verapamil, aPPD did not affect ATPase activity of P-gp suggesting a different mechanism of action. The above results indicate that aPPD, unlike its precursor ginsenosides Rg3 and Rh2, is not a substrate of P-gp. It is also the first time that aPPD has showed a reversible nature of its P-gp inhibition. In addition to its pro-apoptotic nature, aPPD may be a potential new P-gp inhibitor for cancer treatment. PMID- 19291610 TI - Protective effects of gastrodin on lead-induced synaptic plasticity deficits in rat hippocampus. AB - Lead is a well-known toxin in the environment that causes severe damage to the nervous system. Gastrodin is the main bioactive component of Tian ma ( GASTRODIA ELATA Bl.), which is a traditional herbal medicine widely used in eastern Asia. Increasing lines of evidence show that gastrodin has diverse effects, especially neuroprotective effects. In the present study, we investigated whether gastrodin supplementation can rescue impairments of synaptic plasticity produced by developmental lead exposure. We examined three electrophysiological parameters of synaptic plasticity: input/output (I/O) function, paired-pulse facilitation (PPF), and long-term potentiation (LTP) of field excitatory postsynaptic potential (fEPSP) in the hippocampal CA1 region of rats on postnatal day 22 (P22). Our results showed that lead exposure significantly impaired synaptic plasticity in the hippocampal CA1 region and that gastrodin can effectively rescue these lead-induced impairments. Therefore, gastrodin may have potential therapeutic value for lead-induced impairments during human developmental stages. PMID- 19291611 TI - Improved chromatographic assessment of Chinese medicinal products by multi chemical classes analysis. AB - An improved strategy for the quality evaluation of proprietary Chinese medicines (PCM) in complex matrices is presented. It involves the simultaneous separating and displaying of multi-chemical classes and analytes within a single chromatographic elution. The developed fingerprints have the properties of better coverage of multi-chemical classes among the composing herbs. This comprehensive and more facile approach was illustrated for Lemai Keli, a commonly used PCM listed in the Chinese Pharmacopoeia with highly complex analytical matrices. Ten marker compounds belonging to monoterpene glycosides, chalcone glycosides, phenolic acids and phenolic carboxylic aldehyde groups of the analytes together with three common chromatographic peaks were successively separated and displayed within the chromatographic window as a unique HPLC-PAD fingerprint. Rigorous method validation data demonstrated the suitability of the current method for quality assessment purposes in a highly complex PCM matrix. Additional chemometric treatment further demonstrated the uniqueness of the developed fingerprint, with a distinctive chemical profile compared to a highly similar PCM formulation (Kangen Karyu) from which it was considered to be impossible to differentiate when using the conventional single marker approach. PMID- 19291612 TI - Capillary electrophoresis of natural products: current applications and recent advances. AB - Capillary electrophoresis (CE) was introduced as a new analytical technique in the 1970s and rapidly proved to be a powerful tool for the separation and detection of various classes of natural and synthetic compounds. Since the availability of commercially manufactured high-performance instruments, CE represents an interesting alternative to high-pressure liquid chromatography (HPLC), mainly because of its speed and high separation efficiency. In this overview a short description of the basic and widely used CE methods will be given and the applicability of these methods for the analysis of natural products will be discussed. Due to the growing number of publications dealing with CE or CE/MS of secondary plant metabolites, an exhaustive overview of all current applications cannot be given in this contribution. Therefore, the focus of this mini-review will be on the advances and new aspects of recently published CE methods in natural products analysis. PMID- 19291613 TI - New seco-anthraquinone glucosides from Rumex nepalensis. AB - Investigation of the N-BuOH extract of the roots of Rumex nepalensis afforded two new seco-anthraquinone glucosides, nepalensides A and B, along with nine known compounds, torachrysone ( 3), rumexoside, orientaloside, orcinol glucoside, aloesin, lyoniresinol 3 alpha- O- beta- D-glucopyranoside, (-)-epicatechin-3- O gallate, (3,5-dimethoxy-4-hydroxyphenol)-1- O- beta- D-(6- O-galloyl) glucose, and (-)-epicatechin. Their structures were elucidated on the basis of spectroscopic methods. The possible formation of nepalensides A and B is briefly discussed. PMID- 19291614 TI - Effects of maslinic acid, a natural triterpene, on glycogen metabolism in cultured cortical astrocytes. AB - Maslinic acid (2- alpha,3- beta-dihydroxyolean-12-en-28-oic acid) is a triterpenoid compound present in plants of Olea europaea. In the present study, we investigated the effect of maslinic acid on astrocytic glycogen metabolism. Glycogen phosphorylase (GP) activity in homogenates of cultured astrocytes was analyzed, and maslinic acid exhibited GP inhibition with an IC (50) value of 5.7 microM. Moreover, the influence of maslinic acid on glycogen synthesis and glycogenolysis was also investigated. Pre-incubation with maslinic acid dose dependently increased cellular glycogen content and prevented the excessive glycogenolysis induced by norepinephrine. In conclusion, maslinic acid is suggested to be a potent inhibitor of astrocytic glycogen phosphorylase. PMID- 19291615 TI - [Treatment of severe sepsis]. AB - The sepsis syndrome is defined as a clinically suspected infection accompanied by systemic manifestations. The emergence of one or multiple organ dysfunctions is the hallmark of severe sepsis. The significance of sepsis in pulmonary medicine is highlighted by the fact that severe pneumonia is the most prevalent cause of sepsis in non-surgical intensive care medicine and that acute lung injury is among the prominent manifestations of severe sepsis. The formation of major study groups and competence centres during the last decade has made meaningful prospective interventional studies possible that, in turn, have resulted in the development of clinical treatment algorithms. Today, the multidisciplinary treatment of severe sepsis includes haemodynamic, respiratory, endocrine, immune, and coagulation aspects besides the control of the underlying infection. While new concepts of shock treatment and mechanical ventilation have been established by means of confirmatory studies, endocrine interventions as well as immune and anticoagulation procedures are still under investigation. PMID- 19291616 TI - [Dyspnea lusoria]. PMID- 19291617 TI - [Telemonitoring of arm ergometric training in COPD patients. A pilot study]. AB - INTRODUCTION: Regular moderate physical activity in COPD reduces morbidity and improves life expectancy. Many programmes often fail after rehabilitation because regular training at home, possibly caused by a lack of motivation, is not maintained, the performance parameters then decline to the prerehabilitation levels. For cost reasons, a simple procedure is required to measure the training times in the home environment with a daily transmission of telemetric data so that, in the case of decline, advice and assistance can be provided. QUESTIONS: How long is the actual daily training time monitored by telemetry of COPD patients if a target of 20 minutes per day is prescribed by the doctor? Is there a correlation between physical activity at home and the training period? METHOD: We used a device for training the upper extremities with a GSM data transmission module for the transfer of the training times. The physical activity before an exacerbation of COPD and the willingness to modify the training behaviour were determined by questionnaires. 13 patients with COPD (age 69 +/- 9 years, FEV1 48.6 +/- 23.9 %) were investigated prospectively for 6 days under stationary conditions. RESULTS: The mean training period (target 20 min/day) was 16.1 +/- 11.7 min/day. The frequency of physical activity per week correlated with the daily ergometry training period in the clinic (r = 0.59). CONCLUSION: Training supervised by telemonitoring represents a useful way to objectively obtain training times. Ergometric training in combination with online transmission (telemetric monitoring) and regular phone calls could be an inexpensive method to increase physical activity. PMID- 19291618 TI - [Ludolph Brauer--a leader in modern internal medicine and chest medicine]. PMID- 19291619 TI - Giant intrathyroid parathyroid adenoma: a preoperative and intraoperative diagnostic challenge. AB - We describe the case of an unusually large (giant) cystic intrathyroid parathyroid adenoma in a 73-year-old woman who had a 1-year history of hypercalcemia and a 5-year history of an asymptomatic enlargement of the left lobe of the thyroid. This unique case highlights the potential difficulties that can arise in the evaluation of thyroid nodules in patients with hyperparathyroidism. These difficulties were accentuated in this case by the large size of the mass, its intrathyroid location, and cytologic features that were compatible with a lesion of thyroid origin. In some cases, including this one, even a thorough preoperative evaluation that includes fine-needle aspiration biopsy and radiographic and nuclear medicine studies may not allow for a definitive preoperative diagnosis. The histologic overlap between thyroid and parathyroid lesions can also be problematic at the time of intraoperative frozen section evaluation. Intraoperative parathyroid hormone monitoring may be helpful in these difficult cases. PMID- 19291620 TI - Recurrent sudden sensorineural hearing loss sensitive to transtympanic and systemic steroid treatment: case report. AB - Sudden sensorineural hearing loss (SSNHL) is a rare otologic emergency. Recurrence of this pathology is extremely rare, and its incidence remains unknown. We report the case of a previously healthy 28-year-old man with normal findings on bilateral ear examination who presented with severe unilateral SSNHL. Audiometry demonstrated a pure-tone average of 70 dB and a speech discrimination score of 0% in the affected ear. The patient was treated within hours of the onset of symptoms with an intratympanic injection of dexamethasone and then started on a tapered course of oral prednisone. He experienced a full recovery based on repeat audiometric testing 7 days later. However, 1 year later, the patient experienced a repeat episode of SSNHL in the same ear. Management again included an intratympanic injection and an oral steroid course with good results. A complete metabolic and radiographic evaluation failed to identify a cause of the hearing loss. This case represents a highly unusual instance of recurrent SSNHL of unknown etiology. PMID- 19291621 TI - Rosai-Dorfman disease in an Asian Indian woman with classic generalized lymphadenopathy and nasal obstruction: a case report. AB - Rosai-Dorfman disease is rare. When it does occur, it usually affects children, and it has a propensity for whites and blacks as opposed to members of other races. We report the case of a 45-year-old Asian Indian woman who presented with painful masses in the area of the axillary, cervical, and inguinal lymph nodes. She had a decade-long history of tonic-clonic seizures and a recent history of digestive complaints and progressive nasal stuffiness. Endoscopic examination of the left nasal cavity revealed the presence of submucosal bulges along the septum and the lateral wall; hypertrophy of the adenoids was also noted. Histologic analysis of lymph node specimens revealed dilated parenchymal sinuses, germinal activity with infiltration of numerous histiocytes (emperipolesis), and chronic inflammatory cells. The patient was diagnosed with Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) with atypical extranodal involvement (the nasal area). An aggressive course of polychemotherapy thrice daily was initiated, but it had no lasting effect, and the patient died 8 months later of an undetermined cause. Our patient's age at the onset of her disease, her race, and the extranodal involvement make this case of Rosai-Dorfman disease unusual and perhaps unique. PMID- 19291622 TI - Malignant rhabdoid tumor of the parapharyngeal space. AB - Malignant rhabdoid tumor has been a somewhat controversial entity since it was first described in 1978 as a possible sarcomatous variant of Wilms tumor. Eventually, however, it was found to be a distinct neoplastic tumor with histologic characteristics similar to those of rhabdomyosarcoma. Malignant rhabdoid tumors affect children. Clinically, they occur primarily in the kidney, and they behave aggressively. Associated mortality is significant, even with combined-modality treatment regimens. We describe the case of a large extrarenal malignant rhabdoid tumor of the parapharyngeal space with extension to the infratemporal fossa and skull base in a previously healthy 2-year-old girl who had presented with a cervical mass and ipsilateral Horner syndrome. The patient underwent complete surgical extirpation of the lesion and received adjunctive cisplatin chemotherapy and radiation therapy, and she remained disease-free at 9 months of follow-up. Given the age group of the patients that these neoplasms most commonly affect and given the neoplasms' resemblance to rhabdomyosarcoma and other small round-cell tumors of the head and neck, discussion of the associated clinical pathology, imaging characteristics, histopathologic features, and mode of management are of particular importance, especially so in view of the uncommon location of the tumor in this specific case. Such a discussion may help lead to minimization of misdiagnosis and maximization of therapeutic benefit. PMID- 19291623 TI - The thyroid foramen: a case report and literature review of a largely unrecognized laryngeal anomaly. AB - A thyroid foramen is a congenital linear opening located in the lamina of the posterosuperior portion of the thyroid cartilage. Although its overall incidence is reported to be as high as 39%, few practitioners are aware of its existence. We report the case of a 19-year-old man who was involved in a motor vehicle accident. He experienced transient left true vocal fold paresis and what were initially thought to be "bilateral thyroid cartilage fractures" based on computed tomography of the neck performed as part of his trauma evaluation. On further scrutiny, however, the "fractures" were determined to be bilateral thyroid foramina. The presence of the paresis in the setting of these thyroid foramina prompted us to review the literature to examine the significance of thyroid foramina and the vessels and nerves that course through them. The average diameter of a thyroid foramen is approximately 2.5 mm. These openings appear in an oblique orientation, and they occur both unilaterally and bilaterally. In 70% of cases, an anastomotic branch connects the external and internal branches of the superior laryngeal nerve. Embryologically, thyroid foramina are believed to arise from a combined branchial and neurovascular origin. A detailed understanding of laryngeal anatomy, including its variations and their physiologic implications, is important to head and neck surgeons. PMID- 19291624 TI - Surgical treatment is warranted in aggressive central giant cell granuloma: a report of 2 cases. AB - Central giant cell granuloma (CGCG) is a benign but locally destructive lesion of the mandible or maxilla that presents most often in the second and third decades of life. Reports of treatment include curettage or complete en bloc resection and nonsurgical approaches such as intralesional steroid injections, interferon alfa 2a, and calcitonin. We describe the cases of 2 young adults with CGCG of the maxilla involving the palate. The first patient, an 18-year-old man, was initially treated with a partial maxillectomy via a facial degloving approach and a postoperative series of transoral intralesional steroid injections to the site of the residual hard palate disease. The partial maxillectomy achieved a near total debulking of the mass and immediate cosmetic improvement, and the steroid injections resulted in initial regression of the residual disease. However, the patient experienced a recurrence 8 months postoperatively, so a bilateral total inferior maxillectomy via a facial degloving approach was performed with subsequent placement of an obturator. No recurrence was seen 15 months following revision surgery. In view of the incomplete resolution of disease with the use of steroid injections in the first patient, we treated the second patient, a 22-year old woman, with a subtotal maxillectomy without steroid injection. No recurrence of disease was noted 12 months postoperatively. We conclude that a combination of partial surgical resection and intralesional steroid injection may not be sufficient for the treatment of large CGCGs of the maxilla. Complete surgical resection should be considered for the initial treatment of CGCG, particularly in aggressive cases. PMID- 19291625 TI - Resident duty hours: concerns and consequences. PMID- 19291626 TI - Unilateral submandibular gland aplasia: a rare phenomenon. PMID- 19291627 TI - Facial nerve hemangioma of the middle ear. PMID- 19291628 TI - Laryngeal granular cell tumor. PMID- 19291629 TI - Large mixed hiatal hernia and stricture as a cause of dysphagia. PMID- 19291630 TI - Trigeminal trophic syndrome. PMID- 19291631 TI - Sudden-onset life-threatening stridor in an adult caused by a laryngeal ductal cyst. AB - Laryngeal cysts are rare in adults; therefore, the majority of previous reports deal primarily with laryngeal cysts in infants and children. Ductal cysts of the larynx arise from the submucous glands when the ducts of these glands are occluded. They are most commonly located in the vallecula. In this report, we present a rare cause of life-threatening stridor in adults, a large ductal cyst of the larynx that was removed via endoscopic CO(2) laser resection. On the basis of clinical observations during the postoperative period, we conclude that endoscopic CO(2) laser resection is a reliable and safe method for removing cysts of the larynx. It is also a cost-effective procedure that minimizes hospitalization time and the need for tracheotomy. PMID- 19291632 TI - Angiomyomatous hamartoma in a submandibular lymph node: a case report. AB - Angiomyomatous hamartomas have been found almost exclusively in the inguinal and femoral lymph nodes; few reports of these lesions in the head and neck region have been published. We present a case of angiomyomatous hamartoma in the submandibular area, a site that has not been previously reported in the literature. The mass was initially diagnosed as an enlarged lymph node. When it did not regress following broad-spectrum antibiotic treatment, the patient, a 51 year-old woman, underwent an excisional biopsy. Histopathology identified the mass as an angiomyomatous hamartoma. At follow-up 3 years and 9 months postoperatively, the patient exhibited no evidence of recurrence on physical examination and computed tomography. Even though angiomyomatous hamartoma of the head and neck is rare, we suggest that otolaryngologists include it in the differential diagnosis of head and neck masses. PMID- 19291633 TI - Cervical esophagotomy for an impacted denture: a case report. AB - We present the case of a 46-year-old woman with an impacted denture and an impending esophageal perforation. Her family physician initially missed the diagnosis but during a subsequent visit reviewed her x-ray and was able to see the shadow of the denture's wire attachment in her esophagus. The patient was then referred to a tertiary care hospital, where esophagoscopy confirmed the location of the denture, but the surgeon there was unable to remove it. Eighteen days after she had swallowed her denture, she was referred to our hospital. Attempts at removal via rigid esophagoscopy were unsuccessful, but the denture was successfully removed via a cervical esophagotomy. A Gastrograffin swallow performed 1 week postsurgically showed no extravasation of the contrast medium, and subsequent follow-ups were unremarkable. We conclude that cervical esophagotomy is a safe method for removing foreign bodies impacted in the cervical esophagus when they cannot be removed endoscopically. PMID- 19291634 TI - Idiopathic internal carotid artery aneurysm rupture in an infant: a case report. AB - We report a case of idiopathic rupture of an internal carotid artery (ICA) aneurysm in an infant, a finding that we believe has not been previously reported in the literature. The more common causes of isolated injury to the ICA are deep neck space infections and penetrating intraoral trauma. Pseudoaneurysms of the artery are well known to result in delayed hemorrhage. In most cases, the outcome is unfavorable because of the large amount of blood loss. Our case involved a previously healthy 11-month-old girl who presented with massive epistaxis secondary to a ruptured left ICA aneurysm. The patient had no history of any antecedent trauma or infection. She was successfully managed by ligation of the involved ICA in the neck. PMID- 19291635 TI - [Does JUPITER change the daily routine of our practice?]. PMID- 19291636 TI - [Fever in returning travellers]. PMID- 19291637 TI - [CME-Ultrasonography 27]. PMID- 19291638 TI - [Optimised glycaemic control and dose titration with insulin glargine (Lantus) by general practitioners in Switzerland: results of the OPTI-LAN Postmarketing Study]. AB - RATIONALE: National and international societies have published guidelines regarding glycaemic control in type-2 diabetes mellitus. Clinical studies have shown that glycaemic control of type-2 diabetes mellitus can be improved using simple algorithms for titration of insulin Glargine (Lantus). It is unclear, to what degree published guidelines are adopted in daily practice in Switzerland. METHODS: We performed a postmarketing study on the adoption of published guidelines regarding glycaemic control and the use of algorithms for titration of insulin Glargine doses by 77 general practitioners in Switzerland. Data collection was done prospectively over 6 months from September 2005 to September 2007. RESULTS: A total of 317 patients were included with a mean age of 63 +/- 11 years, 137 (43%) were female and 298 (94%) had type-2 diabetes. Average diabetes duration was 8 +/- 6 years. The initial average HbA1C and fasting plasma glucose were 8.9 +/- 1.6% and 10.1 +/- 3.2 mmol/l respectively. This, despite treatment with at least one oral hypoglycaemic agent (83% of patients) and/or insulin (47% of patients). As target the treating physicians chose an HbA1C < or = 7% for 284 patients (90%) and a fasting plasma glucose < or = 7 mmol/l for 245 patients (77%); thresholds that correspond to the guidelines of the Swiss Society of Endocrinology and Diabetes (SSED). The algorithm developed Riddle et al. was used by 33% of treating physicians; whereas 17% of physicians used a schedule proposed by the American Diabetes Association (ADA) and by the European Association for the Study of Diabetes (EASD) that is generally adapted by the patients themselves. During the 6-month treatment with insulin Glargine (Lantus) 196 patients (62%) achieved the SSED guidelines for glycaemic control. On average HbA1C was reduced by 2.1% (8.9 vs. 6.8%) and fasting plasma glucose by 3 mmol/l (10.1 vs. 7.1 mmol/l). On average insulin dose was increased by 14 IU (from 16 to 30 IU). Overall 91% of patients were satisfied with the way insulin doses were adapted. In 64% general wellbeing improved during the treatment period and in 27% it was unchanged. CONCLUSION: In daily practise in Switzerland the recommendations of the SSED are implemented to a high degree. Using insulin Glargine (Lantus) recommended goals of glycaemia can be rapidly and adequately achieved and patient satisfaction can be improved. PMID- 19291639 TI - [Regenerative therapy in cardiology]. AB - The aim of regenerative medicine is the reconstitution of the physiological structure of critically damaged organs. Bone marrow derived stem cells (BMDCs) show promising therapeutic potential. BMDCs are already used in oncology and are ideally suited for regenerative medicine due to their regenerative potential and safety profile. A variety of cells have been contemplated in cardiology, and there is emerging preclinical and clinical data on the feasibility and safety of different cell lines in the setting of acute myocardial infarction and chronic heart failure. In this review, the various concepts and cells will be discussed in further detail. PMID- 19291640 TI - [Elevated transaminases - what to do if everything was done?]. AB - Transaminases, gamma-GT and alcalic phosphatase are classically termed as liver enzymes, however they can be found in almost every organ. Elevated levels of the transaminases ALAT (alanin-aminotransferase) and ASAT (aspartat-aminotransferase) are signs of disturbed permeability of the cells, in which these enzymes can be found. In contrast to ALAT, which is mainly liver-specific, the ASAT is found in other organs as well, e.g. heart and skeletal muscle. At a mild elevation of these enzymes a reevaluation is recommended, however if an elevation persists and is suspicious for a liver disease, a specific work up is necessary. In this manuscript, we discuss often overlooked problems and provide a diagnostic algorithm for the workup of elevated liver enzymes. PMID- 19291641 TI - [Genital warts]. PMID- 19291642 TI - [Sepsis and rhabdomyolysis after holidays in Thailand]. AB - We report a case of severe hypophosphatemia with neurologic, hematologic and musculoskeletal symptoms. Acute hypophosphatemia becomes clinically significant if there is underlying phosphate depletion and can induce a variety of symptoms that can be deleterious. The diagnosis is made by clinical presentation and serum phosphate level. Treatment is promptly warranted, once diagnosis is confirmed. PMID- 19291645 TI - [Answers to questions on hypercalcemia from PRAXIS No. 5]. PMID- 19291646 TI - [CME-Laboratory 10/ Solution on the subject "Disorders of electrolyte and water balance"]. PMID- 19291649 TI - [Epilepsy and epileptic syndromes during the first year of life]. AB - AIM: To analyse the epidemiological characteristics and the relative distribution of the different types of epilepsy and epileptic syndromes during the first year of life. PATIENTS AND METHODS: An analysis was performed of the patient records of all patients with epilepsy diagnosed during their first year of life who were submitted to a developmental check-up in the year 2007. The sample consisted of 60 patients (27 boys and 33 girls). Epidemiological and clinical data were collected, together with the findings from complementary examinations. The diagnostic criteria applied were those of the International League Against Epilepsy. RESULTS: The mean age at the time of diagnosis was 6.3 months. The mean follow-up time was 7.6 years. The aetiology was symptomatic in 40 cases (66.7%), cryptogenic in 16 (26.7%) and idiopathic in four cases (6.7%). Neuroimaging tests detected abnormalities in 34 patients (56.7%). West's syndrome (30%), symptomatic focal epilepsies (23.3%) and epilepsies linked to specific syndromes (16.7%) were the epileptic syndromes with the highest prevalence. Learning disabilities were observed in 82.5% of the children. CONCLUSIONS: Most epilepsies that present during the first year of life are symptomatic and/or cryptogenic, and are accompanied by psychoneurological impairment and/or resistance to therapy, which condition cognitive disorders that are eligible for specialised psycho pedagogical intervention. PMID- 19291650 TI - [Vestibular evoked myogenic potential findings on multiple sclerosis]. AB - INTRODUCTION: Vestibular evoked myogenic potential (VEMP) elicited by acoustic stimulation, has been proposed in the assessment of vestibulospinal pathways. AIM: To research the clinical utility of VEMP on multiple sclerosis (MS). SUBJECTS AND METHODS: Forty-four individuals were evaluated (30 normal state and 14 had MS). The acoustic stimuli were short tone burst (1 kHz, 118 dBNa, rise fall 1 ms, plateau 2 ms) presented through a supra-aural earphone. The stimulation rate was 5 stimuli per second. RESULTS: All the healthy subjects showed a normal VEMP response. In the MS group, abnormal VEMP was recorded among 92.8% of patients and normal in just one case. The P13-N23 increased of prolongation which was the most frequently alteration (57.1%), followed by absence of response among four (28.5%) cases. CONCLUSION: VEMP was considered a good method of diagnostic support of vestibulospinal tract in cases of MS. PMID- 19291651 TI - [A prospective study of neurodevelopment of uninfected children born to human immunodeficiency virus type 1 positive mothers]. AB - INTRODUCTION: The human immunodeficiency virus type 1 (HIV-1) has tropism for the immune and central nervous systems (CNS). Intrauterine exposure to HIV-1 induces immunological alterations, independent of infection that might affect the development of the CNS. Similarly, the intrauterine exposure to antiretrovirals might also affect the neurodevelopment. AIM: To evaluate the neurodevelopment of babies born to HIV-1 positive mothers (exposed) and compare with babies born to HIV-1 negative mothers (unexposed). SUBJECTS AND METHODS: We carried-out an observational prospective study of neurodevelopment of 23 exposed and 20 unexposed children using the infant development scale Bayley-II, and the Denver II test, neurological examination and anthropometric measurements during the first two years of life. RESULTS: None of the exposed babies acquired the infection. At one month of age the exposed babies exhibit normal but statistically lower values in the head circumference, compared to unexposed neonates. No differences were found in the psychomotor development index between both studied groups and exposed babies exhibited a lower mental development index but only at six months of age. The exposed babies exhibited a higher number of alterations during the neurological and Denver-II tests without reaching significant differences. CONCLUSIONS: The results suggest that intrauterine exposure to HIV-1 and to antiretrovirals in uninfected children born to HIV-1 positive mothers does not induce alterations in the neurodevelopment, at least during the first two years of life. PMID- 19291652 TI - [Guillain-Barre syndrome in the paediatric age: epidemiological, clinical and therapeutic profile in a hospital in El Salvador]. AB - INTRODUCTION: Guillain-Barre syndrome (GBS) is an acute polyradiculoneuropathy with, in most cases, an autoimmune aetiology. It manifests as a flaccid motor paralysis, of a distal ascending type, accompanied by areflexia with or without sensory alterations. AIM: To determine the clinical and therapeutic epidemiological profile of patients diagnosed as suffering from GBS who were attended at the Benjamin Bloom Children's Hospital in El Salvador, between 1st January 1997 and 31st December 2004. PATIENTS AND METHODS: We performed an observation-based, descriptive, cross-sectional study that included 414 cases. The following data were evaluated: age distribution, sex, age, history of infections, clinical manifestations, results of the lumbar puncture, electrophysiological studies and treatment that was received. RESULTS: A predominance of males was observed, together with a more frequent presentation in school-age children. The number of cases increased in the third quarter of each of the years included in the study. The most prevalent event in the history of infections was acute respiratory infection. And the most frequent clinical presentation was ascending weakness. Albuminocytological dissociation was detected in 88% of patients who had undergone lumbar puncture. Acute inflammatory demyelinating polyneuropathy predominated in 88.3% of cases. Sixty per cent of patients had to be admitted to the intensive care unit. In most cases treatment consisted in intravenous administration of immunoglobulin. Complications occurred in 30.5% of cases and the mortality rate was 2.9%. CONCLUSIONS: GBS occurs predominantly in the rainiest season of the year, which is related to an increase in the number of infections in the upper respiratory tract and gastroenteric infections that give rise to a predisposition to present secular outbreaks of the disease. PMID- 19291653 TI - [Thrombosis of the cavernous sinus following prone-position spinal surgery in a patient with factor V Leiden]. AB - INTRODUCTION: Non-infectious thrombosis of the cavernous sinus has sometimes been reported as a complication following neurosurgical procedures and, in one case, after carrying out the operation in a prone position. Factor V Leiden is a genetic risk factor for presenting an intracranial venous thrombotic disease. We report the case of a patient who suffered thrombosis of the cavernous sinus following prolonged surgery in the prone position and in whom a mutation of factor V Leiden in a heterozygotic state was discovered. CASE REPORT: A 64-year old male, with arterial hypertension as the only known vascular risk factor, who, after prolonged surgery in a prone position, presented amaurosis in the left eye accompanied some hours later by ocular pain, conjunctival ecchymosis, proptosis and abolition of extrinsic ocular mobility. An angiography scan confirmed the existence of thrombosis in the cavernous sinus. Treatment was established with low-molecular-weight heparin without the occurrence of any other kinds of complications. The patient gradually recovered extrinsic ocular mobility but not visual acuity. A heterozygotic mutation for factor V Leiden was found in a hypercoagulability study that was later performed. CONCLUSIONS: The coexistence of a risk factor for presenting a venous thrombotic disease and a mechanical phenomenon, venous statis, due to the posture adopted for the surgical intervention together account for the complication presented by the patient. PMID- 19291654 TI - [Idiopathic hypertrophic pachymeningitis: a case report]. AB - INTRODUCTION: Hypertrophic pachymeningitis is a clinical condition that is caused by a diffuse or localised thickening of the dura mater. It predominantly affects males and manifests as chronic headache, with or without association to neurological manifestations, such as paralysis of the cranial nerves, cerebellar ataxia and neuro-ophthalmic complications. CASE REPORT: A 61-year-old male, with no relevant past history, who, one month before admission, had begun to suffer from right frontotemporal headache that irradiated to the ipsilateral orbital region and was more pronounced at night. A week later he was affected by a decrease in visual acuity in the right eye and two weeks later he noted the presence of right palpebral ptosis, while the headaches increased. The examination showed: right palpebral ptosis with global ophthalmoparesis with predominance of adduction and abduction, and diminished photomotor reflex in the right eye. The visual acuity of the right eye was reduced and the palpebral fissure was 0 in the right eye. The fundus oculi was normal. Infectious and non infectious causations of meningitis were precluded. Magnetic resonance imaging revealed a diffuse thickening of the supratentorial and infratentorial meninges, as well as diffuse uptake of the paramagnetic substance; thickening of the mucus in both paranasal maxillary sinuses was also observed. A meningeal biopsy study confirmed the existence of hypertrophic pachymeningitis. Treatment was established with prednisone and the clinical symptoms improved. CONCLUSIONS: Idiopathic hypertrophic pachymeningitis is an underdiagnosed condition that must be taken into consideration in cases of patients with a history of subacute or chronic meningitis in which infectious and non-infectious causations have been precluded, and high-dose steroid treatment must be established. PMID- 19291655 TI - [Mechanisms of secondary degeneration in the central nervous system during acute neural disorders and white matter damage]. AB - INTRODUCTION: Acute neurodegenerative diseases, including stroke and traumatic brain and spinal cord injury, possess an elevated worldwide incidence. Two distinct lesive patterns can be identified after these destructive events: primary damage, an early consequence of the primary pathological event, and secondary neural degeneration (SND), a group of pathological events inducing late degeneration in cells not or even only partially affected by the primary damage. This pathological mechanism is an important contributing factor for functional deficits and target for therapeutic approaches. Several factors are involved on the SND etiology, including excitotoxicity, inflammation, and oxidative stress. AIM: To review the main mechanisms underlying the SND occurring after acute neural disorders. DEVELOPMENT: The more recent findings about the eliciting processes of SND degeneration are discussed, as well as their significance to degeneration of white matter tracts. CONCLUSIONS: The characterization of the events underlying SND is of fundamental importance for the development of new therapeutic approaches effective enough to decrease the functional deficits, contributing to the improvement of the quality of life of people suffering neurological diseases. These therapeutic approaches must be validated in experimental models of both brain and spinal cord diseases, which effectively simulate human neural disorders protecting both gray and white matters for a better neuroprotective efficacy. PMID- 19291656 TI - [Paraneoplastic limbic encephalitis: a difficult-to-diagnose condition]. AB - INTRODUCTION: Paraneoplastic syndromes can be defined as manifestations in distant places of tumors or metastasis, which are not related with tumoral growth. Most of these syndromes are caused by substances secreted by the tumor, that mimic natural hormones, or interfere with plasma proteins. DEVELOPMENT: The rate of paraneoplastic syndromes with neurological manifestations is less than 0.5/100,000 per year, and affect about 0.01% of cancer patients. The pathogenesis of neurological paraneoplastic syndromes is attributed to humoral autoimmunity, due to the existence of a great variety of antibodies in relationship with the neurological alterations associated. Nevertheless, the absence of antibodies does not exclude a neurological paraneoplastic syndromes, just as antibodies may be found without a neurological paraneoplastic syndrome. The characteristic symptoms of paraneoplastic limbic encephalitis are confusion of acute onset, mood changes, hallucinations, loss of short term memory, and seizures; these symptoms generally develop in days or weeks, but may present suddenly. Image studies, cerebral spinal fluid evaluation, and serologic tests are the most useful in diagnosing a neurological paraneoplastic syndrome. The treatment requires two different approaches. The first one is through the suppression of the immune response generated by neurological damage. The second, is by removing the tumor as the source of the antigen. The latter is often the only effective treatment. CONCLUSIONS: The paraneoplastic limbic encephalitis is an unusual and hard to diagnose entity, which can easily be confused with psychiatric problems. An early diagnosis and treatment is very important to avoid nonreversible neuronal damage. PMID- 19291657 TI - [Connectionist simulation in neuropsychology]. AB - INTRODUCTION: The expression 'connectionist neuropsychology' has been applied since the early 1990s to designate an emerging area of research that uses artificial neural networks in an attempt to simulate some of the cognitive disorders that result from different kinds of brain injury. Although some of the models generated by this type of simulation offer a number of advantages over the classic models of information processing, this approach is still not very well known among Spanish researchers and health professionals. AIMS: To make this important research tool more widely known and to review the advantages and shortcomings of a series of particular models. DEVELOPMENT: After a brief introduction to the fundamental principles of connectionist simulation, some of the most important models involving aphasias, dyslexias, agnosias and apraxias are outlined. CONCLUSIONS: Despite their limitations, the models proposed by connectionist neuropsychology seem to be far more accurate and specific than the classic ones. Furthermore, they are easy to handle and make it possible to get much closer to the complex reality of these disorders. This type of research allows different kinds of brain injury to be modelled while also enabling researchers to explore brain functions that have remained unknown up till now, either for ethical or practical reasons. It therefore represents a source of inspiration both for designing experimental research studies and for developing new rehabilitation procedures. PMID- 19291658 TI - [Aretaeus of Cappadocia (2nd century AD) and the earliest neurological descriptions]. AB - INTRODUCTION AND DEVELOPMENT: Aretaeus of Cappadocia, who was possibly a contemporary of Galen, is considered to have been one of the best clinical physicians of the Ancient World. Nothing is known of his biography, except for constant references to his probable place of birth, Cappadocia. His Extant Works, one of the most important and influential treaties on Greco-Roman medicine, has survived to our days (although it is incomplete). It consists of eight books, in which he gives an orderly and precise account of the aetiology, symptomatology and therapeutics of acute and chronic diseases. Several chapters, possibly devoted to neurological matters (phrenitis, lethargy, wasting and apoplexy), are missing from Book I. Book III includes matters such as headaches, scotoma, epilepsy, melancholy, madness and paralysis. CONCLUSIONS: Aretaeus has always stood out for his capacity for observation as well as the thoroughness of his nosographic descriptions, which in many cases, like migraine or epilepsy, has led to his accounts being considered as seminal works. As the leading representative of the pneumatic school as far as aetiology is concerned, Aretaeus added a fifth element to the classical Greek stoichiology, pneuma (spirit), which permeates everything and, when altered, gives rise to diseases. PMID- 19291659 TI - [Prognosis and functional neuroimaging in a case of methanol intoxication]. PMID- 19291660 TI - [Polypills in the secondary prevention of strokes]. PMID- 19291661 TI - [Amyotrophic lateral sclerosis and pregnancy]. PMID- 19291662 TI - [Monkey sign in pontine myelinolysis]. PMID- 19291663 TI - [Gabapentin in the treatment of ocular neuromyotonia]. PMID- 19291664 TI - [Ischemic stroke due to transient left ventricular apical ballooning in women with epilepsy debut]. PMID- 19291665 TI - [Cyanide and carbon monoxide in intoxication by smoke in a fire]. PMID- 19291667 TI - Self-assembled nanogel of hydrophobized dendritic dextrin for protein delivery. AB - Highly branched cyclic dextrin derivatives (CH-CDex) that are partly substituted with cholesterol groups have been synthesized. The CH-CDex forms monodisperse and stable nanogels with a hydrodynamic radii of approximately 10 nm by the self assembly of 4-6 CH-CDex macromolecules in water. The CH-CDex nanogels spontaneously trap 10-16 molecules of fluorescein isothiocyanate-labeled insulin (FITC-Ins). The complex shows high colloidal stability: no dissociation of trapped insulin is observed after at least 1 month in phosphate buffer (0.1 M, pH 8.0). In the presence of bovine serum albumin (BSA, 50 mg . mL(-1)), which is a model blood system, the FITC-Ins trapped in the nanogels is continuously released ( approximately 20% at 12 h) without burst release. The high-density nanogel structure derived from the highly branched CDex significantly affects the stability of the nanogel-protein complex. PMID- 19291668 TI - Non-isothermal kinetics of hard alpha-keratin thermal denaturation. AB - DSC measurements carried out at different heating rates were used for the kinetic analysis of the endothermic process assigned to the denaturation of the helical material from human hair in water excess. We found that the kinetic mechanism is autocatalytic and that the value of the activation energy is close to disulphide bond scission rather than to protein denaturation. This allowed us to propose a multistep mechanism for the thermal denaturation of hard alpha-keratins in water excess that relies on the 3-phase model which describes their structure. The limiting step of the thermal denaturation process is then the scission of S-S bonds between the main morphological components, namely IF and matrix (IFAP). The theoretical proposed model shows a good agreement with the experimental recorded data. PMID- 19291669 TI - Computer-assisted therapy in orbital and mid-facial reconstructions. AB - BACKGROUND: Management of orbital and mid-facial fractures requires a thorough ophthalmic evaluation and precise imaging. A principle goal of therapy is to anatomically reduce fracture segments and to restore a normal orbital volume as soon as possible. Diagnostic advances such as new surgical and imaging techniques have dramatically improved both the functional and aesthetic outcome of reconstructions. METHODS: Orbital reconstruction is performed using computer assisted navigation. This technique makes preoperative simulation by mirroring the unaffected side onto the affected side. RESULTS: Results from computer assisted navigation application to both primary and secondary orbital and mid facial reconstruction are shown. CONCLUSION: Navigation technique it offers significant advantages in both primary and secondary reconstructions. Navigation facilitates reconstruction in unilateral defects through mirroring techniques, and in bilateral defects by importing virtual models from standard CT datasets improving the software tool to fulfil the need for maxillofacial surgery reconstruction. PMID- 19291670 TI - A mass spectrometric study on meloxicam metabolism in horses and the fungus Cunninghamella elegans, and the relevance of this microbial system as a model of drug metabolism in the horse. AB - This paper describes a study where the metabolism of the non-steroidal anti inflammatory drug meloxicam was investigated in six horses and in the filamentous fungus Cunninghamella elegans. The metabolites identified were compared between the species, and then the fungus was used to produce larger amounts of the metabolites for future use as reference material. C. elegans proved to be a good model of phase I meloxicam metabolism in horses since all four metabolites found were the same in both species. Apart from the two main metabolites, 5' hydroxymethylmeloxicam and 5'-carboxymeloxicam, a second isomer of hydroxymeloxicam and dihydroxylated meloxicam were detected for the first time in horse urine and the microbial incubations. Phase II metabolites were not discovered in the C. elegans samples but hydroxymeloxicam glucuronide was detected intact in horse urine for the first time in this study. Urine from six horses was further analyzed in a semi-quantitative sense and 5' hydroxymethylmeloxicam gave peaks with much higher intensity compared to the parent drug and the other metabolites, and was detected for at least 14 days after the last given dose in some of the horses. From the results presented in this article, we suggest that analytical methods developed for the detection of meloxicam in horse urine after prohibited use should focus on the 5' hydroxymethyl metabolite and that C. elegans can be used to produce large amounts of this metabolite for potential future use as a reference compound. PMID- 19291671 TI - The female gonad in two species of Microplana (Platyhelminthes, Tricladida, Rhynchodemidae): ultrastructural and cytochemical investigations. AB - The female gonad of the land planarians Microplana scharffi and Microplana terrestris consists of two small germaria located ventrally in the anterior third of the body and of two ventro-lateral rows of oblong vitelline follicles distributed between the intestinal pouches. Both these structures are enveloped by a tunica composed of an outer extracellular lamina and an inner sheath of accessory cells. Oocyte maturation is characterized by the appearance of chromatoid bodies and the development of endoplasmic reticulum and Golgi complexes. These organelles appear to be correlated with the production of egg granules with a fenestrated/granular content of medium electron density, about 4 5 mum in diameter, which remain dispersed in the ooplasm of mature oocytes. On the basis of cytochemical tests showing their glycoprotein composition, and their localization in mature oocytes, these egg granules have been interpreted as yolk. In the vitelline follicles, vitellocytes show the typical features of secretory cells with well-developed rough endoplasmic reticulum and Golgi complexes involved in the production of eggshell globules and yolk. The eggshell globules, which appear to arise from repeated coalescences of two types of Golgi-derived vesicles, contain polyphenols and, when completely mature, they measure about 1 1,2 mum in diameter and show a meandering/concentric content pattern as is typical of the situation observed in most Proseriata and Tricladida. Mature vitellocytes also contain a large amount of glycogen and lipids as further reserve material. On the basis of the ultrastructural features of the female gonad and in relation to the current literature the two species of rhynchodemids investigated appear to be closely related to the freshwater planarians belonging to the family Dugesiidae. PMID- 19291672 TI - Development of transient head cavities during early organogenesis of the Nile Crocodile (Crocodylus niloticus). AB - Three consecutive pairs of head cavities (premandibular, mandibular, and hyoid) found in elasmobranchs have been considered as remnants of preotic 'head' somites serial homologues of the myotomic compartments of trunk somites that give rise to the extraoccular musculature. Here, we study a more derived vertebrate, and show that cavitation is more complex in the head of Crocodylus niloticus, than just the occurrence of three pairs of cavities. Apart from the premandibular cavities, paired satellite microcavities, and unpaired extrapremandibular microcavities are recognized in the prechordal region as well. We observed that several developmental phenomena occur at the same time as the formation of the head cavities (premandibular, satellite, extrapremandibular, mandibular, and hyoid) appear temporarily in the crocodile embryo. These are 1) rapid growth of the optic stalk and inflation of the optic vesicle; 2) release of the intimate topographical relationships between the neural tube, notochord and oral gut; 3) tendency of the prechordal mesenchyme to follow the curvature of the forebrain; and 4) proliferation of the prechordal mesenchyme. On the basis of volumetric characters, only the hyoid cavity and hyoid condensation is comparable to the trunk somitocoel and somite, respectively. PMID- 19291673 TI - Effect of the content of unmethylated CpG dinucleotides in plasmid DNA on the sustainability of transgene expression. AB - BACKGROUND: Nonviral gene transfer generally suffers from short-term expression of transgenes. We have previously demonstrated that plasmids with reduced CpG content exhibited a more prolonged expression of murine interferon (IFN)-beta or IFN-gamma, which was effective in inhibiting metastatic tumor growth. A further extension of the duration of transgene expression could be achieved by controlling the number and location of CpG motifs in plasmid DNA. METHODS: Luciferase-expressing plasmids with differing CpG content were injected into the tail vein of mice by the hydrodynamic injection method. The effects of CpG content on the duration of transgene expression were examined, focusing on cytosine methylation and pro-inflammatory cytokines. Based on the findings, IFN gamma-expressing plasmids were constructed and their transgene expression and inhibitory effect on pulmonary metastasis were evaluated. RESULTS: Plasmids with a few CpG motifs showed a prolonged luciferase activity in the liver. Methylation of CpG motifs in plasmids reduced the expression and the extent of this reduction was greater for plasmids with a high CpG content. Pro-inflammatory cytokines hardly affected the expression. pCpG-Mu gamma, the IFN-gamma-expressing plasmid, which contains 20 CpG motifs only in the cDNA region, exhibited a sustained IFN gamma concentration at therapeutic levels, and had a great inhibitory effect on the pulmonary metastasis of tumor cells. CONCLUSIONS: The duration of transgene expression of IFN-gamma was successfully increased by reducing the CpG content of IFN-expressing plasmid vector, which resulted in an increased anticancer activity of IFN gene transfer. PMID- 19291674 TI - Simultaneous quantification of three major triterpenoids in radix asteris by high performance liquid chromatography with evaporative light scattering detection. AB - INTRODUCTION: Radix asteris, with triterpenoids as its main pharmacological effective compounds, has been widely used for moistening the lung, dispersing phlegm and relieving cough. Quantification of the triterpenoids is important for the quality control of Radix asteris. OBJECTIVE: To establish a high-performance liquid chromatography method with evaporative light scattering detection for simultaneous determination of three major triterpenoids, shionone, friedelin and epi-friedelinol, in Radix asteris. METHODOLOGY: The optimal chromatographic conditions were achieved on an RP(18) column with gradient elution by acetonitrile and 0.05% acetic acid in 22 min with ELSD set at an evaporating temperature of 40 degrees C. Validation of the method included tests of linearity, sensitivity, precision, repeatability, stability and accuracy. RESULTS: All calibration curves showed good linear regression (r(2) > 0.9991) within test ranges. The established method showed good precision and accuracy with overall intra-day and inter-day variations of 1.61-2.97 and 1.74-2.42%, respectively, and overall recoveries of 97.35-101.13% for the three compounds analysed. CONCLUSION: The method developed was successfully applied to quantify the main triterpenoids in 14 Radix asteris samples. PMID- 19291675 TI - Qualitative and quantitative determination of hydrolysable tannins and other polyphenols in herbal products from meadowsweet and dog rose. AB - INTRODUCTION: Meadowsweet and dog rose are used as herbal medicines and foodstuffs. Their biological and pharmacological properties are mainly attributed to vitamins, flavonoids and tannins known to be effective antioxidants. OBJECTIVE: To evaluate the distribution of hydrolysable tannins and other polyphenols in commercially available products of Filipendula ulmaria and Rosa canina. METHODOLOGY: In order to evaluate the distribution of polyphenols, the isolation and structure elucidation of six ellagitannins (tellimagrandins I and II, rugosins A, B, D and E) along with other well-known compounds was performed. Further high-performance chromatographic methods (HPLC, HPTLC) were developed for qualitative and quantitative studies of phenolic acids, flavonoids and tannins in meadowsweet flowers and dog rose fruits. The analysis of linearity, limits of detection and quantification, precision and accuracy showed that the HPLC assay was sufficiently precise for content evaluation. Selection of a suitable solvent and optimisation of the sample extraction procedure were also carried out. RESULTS: Amounts of more than 20 polyphenols were estimated in the studied pharmaceuticals (27 products of meadowsweet and 10 products of dog rose) using a Chromolith Performance RP-18e column and UV detection. Methyl gallate 3-O-beta glucoside was identified for the first time in dog rose, while rugosins A, B and E are new meadowsweet constituents. CONCLUSION: The results showed that meadowsweet flowers and rose hips with seeds yielded 55.53-124.8 and 0.43-1.26 mg/g of ellagitannins, respectively. The sum of detected polyphenols was 83.88 165.68 mg/g for Filipendulae ulmariae flos and 1.21-2.66 mg/g for Rosae pseudo fructus cum fructibus. PMID- 19291676 TI - A hypoxic inducible factor-1 alpha hybrid enhances collateral development and reduces vascular leakage in diabetic rats. AB - BACKGROUND: Diabetes mellitus is a common comorbidity of atherosclerosis. Hypoxia inducible factor-1 (HIF-1) is the master regulator of the angiogenic response to hypoxia. METHODS: We studied the effects of adenoviral vectors expressing a constitutively active HIF-1 alpha hybrid (Ad2/HIF-1 alpha/VP16) or vascular endothelial growth factor (Ad2/VEGF) on collateral development and vascular leakiness in a diabetic rat model of hindlimb ischemia. RESULTS: After the removal of the right femoral artery, the mRNA levels of VEGF, angiopoietin-1 and angiopietin-4 in the calf muscles, as measured by Taqman reverse transcriptase polymerase chain reaction, were transiently elevated in Zucker lean (ZL) but not Zucker diabetic fatty (ZDF) rats. The angiographic score, as determined by post mortem angiography, was significantly lower in ZDF animals 35 days after surgery compared to their ZL counterparts. In separate animals, intramuscular injection of Ad2/HIF-1a/VP16 and Ad/2VEGF into the thigh muscles significantly increased the angiographic score and capillary density 21 and 35 days after the injection compared to Ad2/CMVEV (a vector expressing no transgene) or vehicle. After the injection of Ad2/CMVEV or vehicle, the Evans-blue dye content in the thigh muscles was significantly higher in ZDF rats than their ZL counterparts. Ad2/HIF 1 alpha/VP16 but not Ad2/VEGF reduced tissue Evans blue dye content. CONCLUSIONS: The endogenous angiogenic response to ischemia was impaired in ZDF rats, possibly due to down-regulation of angiogenic factors. Ad2/HIF-1 alpha/VP16 enhanced collateral development and reduced vascular leakage in the ischemic hindlimb of ZDF rats indicating that hybrid HIF-1 alpha angiogenic therapy may be efficacious for peripheral vascular disease with a diabetic comorbidity. PMID- 19291677 TI - Temporal metabolomic analysis of o-glucoside phenolic compounds and their aglycone forms in olive tree and derived materials. AB - INTRODUCTION: Maturity is one of the most important factors associated with evaluation of the quality of fruit and vegetables. In olive oil, maturation plays a key role in the kinetics of biosynthetic pathways of the secondary metabolism. One of the most relevant pathways is that catalysed by beta-glucosidases, which are involved in olive oil debittering. Therefore, the knowledge of this influence can be of particular interest for olive oil industry. OBJECTIVE: To monitor the profile of O-glucoside phenols and their aglycone forms in olive oil, alperujo (the semisolid residue resulting in the production of olive oil), stones, leaves and branches in order to interpret its evolution according to the sample and the period of the season (October, when olive drupes are green; December, when these drupes are green-purple; and February, when they are mostly black). METHODOLOGY: Targeted phenols were extracted by previously optimised methods assisted by auxiliary energies (i.e. ultrasound, microwaves or superheated liquids) according to the characteristics of the sample. The analysis was carried out by liquid chromatography-tandem mass spectrometry with a triple quadrupole mass detector. Highly selective identification and sensitive determination of metabolites was performed in multiple-reaction monitoring mode. Statistical analysis to evaluate differences in the profile of the target compounds was based on principal compounds analysis. RESULTS: The evolution of the analytes concentration is strongly related to the role of beta-glucosidases. An explanation for this evolution in olive oil, alperujo and stones is given by relation to the industrial process for olive oil production. For leaves and small branches, the concentration was practically constant over the season, which was foreseeable because of the perennial character of olive trees. Leaves and branches were found to be highly concentrated in O-glucoside derivatives, demonstrating their capacity for phenolic compounds storage. CONCLUSIONS: Targeted metabolomic profiling has proved a useful tool to monitor O-glucoside phenolic compounds and their aglycone forms in olive materials. The profile of target compounds enables interpretion of their evolution according to the olive material and the period of the season. PMID- 19291678 TI - Characterisation of tannin-containing herbal drugs by HPLC. AB - INTRODUCTION: Herbal drugs containing tannins are characterised in the European Pharmacopoeia by their tannin content analysed by the old, nonspecific, colorimetric Folin-Ciocalteu method. The result of the analysis is a single figure relating to the content of tannins, but this does not provide much information on the identity or status of the herbal drug. OBJECTIVE: In the present paper methods for obtaining more detailed information of the constituents in these herbal drugs are described. METHODOLOGY: The methods developed are based on a reversed-phase gradient HPLC system coupled to DAD, fluorescence, electrochemical and MS detectors. RESULTS: The HPLC system developed provides characteristic fingerprints of the herbal drugs when using UV detection at 250 nm. The fingerprints may be used for identification of tannin-containing herbal drugs. Methanolysis of the herbal drug generated methyl gallate and ellagic acid, which were analysed in the HPLC system. The molar ratio between methyl gallate and ellagic acid may also be used for the identification of herbal drugs. CONCLUSIONS: An HPLC system equipped with selective detectors was shown to be valuable in the identification of herbal tannin. Most promising was fingerprinting using UV detection, but methanolysis followed by HPLC also proved useful. PMID- 19291679 TI - Field experiments to assess approaches for spray drift incident investigation. AB - BACKGROUND: Spray trials were conducted to determine the variation in primary spray drift deposition between trials under very similar conditions, in order to assess the feasibility of developing a computational tool to aid post-event investigations of pesticide spray incidents. Pesticide deposition was examined by analysis of filter paper and vegetation samples. RESULTS: Considerable variation in the drift profile was found. The overall estimate of the spray drift decay term was -1.13 (95% confidence interval -1.02 to -1.24), with statistically significant differences between plots. Variation in the drift profile between neighbouring essentially identical plots indicates the variation in deposition that might be expected over small distances. Vegetation samples were found to have considerably lower capture efficiency than filter papers. Importantly, degradation of pesticides was found to have little effect on the pesticide drift profile over a 14 day period. CONCLUSIONS: The levels of spatial variation in spray drift deposits between runs and plots observed in this study suggest serious limitations to the inferences that may be drawn from limited numbers of post-incident samples. In particular, they would limit inferences about the spray conditions that could be drawn from an estimate of the drift profile derived from limited post-incident samples. PMID- 19291680 TI - Effectiveness of isotopically labelled and non-isotopically labelled internal standards in the gas chromatography/mass spectrometry analysis of sulfur compounds in wines: use of a statistically based matrix comprehensive approach. AB - The effectiveness of isotopically and non-isotopically labelled internal standards in reducing matrix-induced effects is evaluated. The question is addressed in the quantitative analysis by gas chromatography/mass spectrometry (GC/MS) of dimethyl sulphide, chosen as a typical example of volatile sulphur compounds, in wine matrices. When matrix/run effects are not cancelled out the use of a variance component model (VCM) to handle the linear calibrations obtained by regression technique is successful. The method implies the estimation of an overall calibration straight line, which properly takes into account the uncertainty due to different matrices, the calibration run and the measurement error, making the use of an isotopically labelled internal standard not necessary. The obtained results show that the benefits of lowering times and costs for routine analyses compensate for the small increase in uncertainty in the concentration values obtained in the regression analysis and the slight increase in the detection limit. PMID- 19291681 TI - Morphological changes during postembryonic development in two species of neotropical harvestmen (Opiliones, Laniatores, Cranaidae). AB - Morphological changes during postembryonic development in the Cranaidae are described on the basis of the examination of an incomplete series of larvae, nymphs, and adults of Phareicranaus calcariferus and Santinezia serratotibialis. The life histories of these species are hypothesized to consist of six nymphal stages, featuring the appearance of secondary male sexual characteristics in the antepenultimate nymph (N5). Color and body shape change dramatically during development. Growth rates for nymphs based upon leg measurements were similar for both species. In S. serratotibialis, the greatest increase in leg size occurred from larva to 1st nymph. The tarsomeres of legs I-IV varied by 1-2 segments per leg for each nymph stage, with the number of tarsal segments increased by 1-2 segments at each stage. Adults had nearly twice as many tarsomeres on leg II than other legs. Ontogenetic changes were observed in the armature of the proximal cheliceral segment, ocularium, pedipalp, opisthosoma, distitarsus III and IV, and leg IV. Morphological changes in postembryonic development in cranaid harvestmen are similar to those reported for other Laniatores. PMID- 19291682 TI - In vivo bioactivity of DNA-based coatings: an experimental study in rats. AB - DNA-based coatings possess beneficial properties useful for medical and dental implants. The present study evaluated the potential in vivo bioactivity of DNA based coatings, either or not pretreated in simulated body fluid (SBF). DNA-based coatings were generated on titanium cylinders using layer-by-layer deposition, with bis-ureido-surfactant as the cationic component and DNA as the anionic component. Noncoated titanium implants and CaP-coated implants served as controls. A total of 80 implants, divided in four experimental groups (n = 10) were implanted unilaterally into the lateral femoral condyles of 80 rats. After implantation periods of 1 and 4 weeks, the bone-to-implant contact and bone volume around the implants were determined histomorphometrically. The results of this study showed that DNA-based coatings and CaP coatings increased bone-to implant contact after 1 week compared to noncoated controls. After 4 weeks of implantation, bone-to-implant contact increased significantly for SBF pretreated DNA coatings and CaP coatings, whereas DNA-coated implants showed no additional effect. The bone-to-implant contact of noncoated controls increased to the level of the DNA-coated implants. Consequently, this study demonstrates that DNA-based coatings are histocompatible and favor early bone responses. SBF-pretreated DNA based coatings were found to increase both early and late peri-implant bone responses. PMID- 19291683 TI - The effect on bone growth enhancement of implant coatings with hydroxyapatite and collagen deposited electrochemically and by plasma spray. AB - Skeletal bone consists of hydroxyapatite (HA) [Ca(10)(PO(4))(6)(OH)(2)] and collagen type I, both of which are osseoconductive. The goal of osseointegration of orthopedic and dental implants is the rapid achievement of a mechanically stable long-lasting fixation between bone and an implant surface. In this study, we evaluated the mechanical fixation and tissue distribution surrounding implants coated with three surfaces: plasma-sprayed HA coating, thinner coating of electrochemical-assisted deposition of HA, and an identical thin coating with a top layer of mineralized collagen. Uncoated plasma-sprayed titanium (Ti-6Al-4V) served as negative control. The electrochemical-assisted deposition was performed near physiological conditions. We used a canine experimental joint replacement model with four cylindrical implants (one of each treatment group) inserted in the humeri cancellous metaphyseal bone in a 1 mm gap. Observation time was 4 weeks. The mechanical fixation was quantified by push-out test to failure, and the peri-implant tissue formation by histomorphometric evaluation. HA coatings deposited by plasma spray technique or electrochemically, increased the mechanical fixation and bone ongrowth, but there was no statistical difference between the individual HA applications. Addition of collagen to the mineralized phase of the coating to create a more bone natural surface did not improve the osseoconductive effect of HA. PMID- 19291684 TI - Biological response of osteoblast-like UMR-106 cells to the modified PHBV matrix- effects of porosity and collagen dip coating. AB - In this study, we related porosity and collagen coating of poly(3-hydroxybutyrate co-3-hydroxyvalerate) (PHBV) scaffold to the degree of cell proliferation on the engineered PHBV scaffold. Based on the [3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2h-tetrazolium, inner salt] (MTS) assay, we established that UMR-106 cell proliferation is maximum in collagen-coated porous PHBV film followed by porous PHBV film and least in nonporous PHBV film. RT-PCR analysis of the proliferated cells on tissue culture polystyrene (TCPS) and porous and nonporous PHBV scaffolds revealed that the proliferated cells retained their osteoblastic phenotype characteristics. Atomic absorption analysis was performed to measure the extent of calcium conversion by the cells grown on PHBV and TCPS. The calcium content of the culture media was used to indirectly measure the mineralization ability of the cells. The extent of calcium conversion by the cells was found to depend on the incubation time. Based on the results of the study, modified PHBV matrix seems to be a suitable matrix candidate for bone tissue engineering application. PMID- 19291685 TI - Redundancy of the effect of TGFbeta1 and beta-NGF on the second meiotic division of rat spermatocytes. AB - We have previously shown that in cocultures of late pachytene/diplotene spermatocytes (PS/DS) with Sertoli cells, beta-nerve growth factor (beta-NGF) or transforming growth factor (TGFbeta1) regulates the second meiotic division by blocking secondary spermatocytes in metaphase II, and thereby lowers round spermatid formation. In this study, we raised the question if beta-NGF and TGFbeta1 have additional or redundant effects on this step. Hence, we addressed the effect of beta-NGF in combination with TGFbeta1, as compared to those of beta NGF or TGFbeta1 separately, on the completion of meiosis by rat late PS/DS. Identification and counting of meiotic cells were performed by cytological methods. Under our culture conditions, some late PS/DS differentiated into round spermatids. When beta-NGF in combination with TGFbeta1 was added to the culture medium, the numbers of PS/DS and of secondary spermatocytes were not modified by the treatment. By contrast, the number of round spermatids was about 2-fold lower in treated cultures than in basal cultures, and an increase in metaphase II, but not metaphase I, over basal values was observed. Similar results were found when either beta-NGF or TGFbeta1 was added to the culture medium. These results indicate a redundancy between beta-NGF and TGFbeta1 on the negative regulation of the second meiotic division of rat spermatocytes. This suggests that beta-NGF and TGFbeta1 share some common pathway of regulation of this step. In addition, these results reinforce the view that the adjustment of the number of round spermatids supported by Sertoli cells is a key event of the spermatogenic process. PMID- 19291686 TI - Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri. AB - Ultrastructural aspects of citrus canker development were investigated in nonwounded leaves of citrus species by transmission electron microscopy (TEM). A susceptible species Mexican lime and a resistant species Yuzu were spray inoculated with a virulent strain of Xanthomonas citri pv. citri. Initial symptoms occurred on Mexican lime approximately 9 days after inoculation, whereas they appeared on Yuzu mostly 11 days after inoculation. In Mexican lime leaves, the bacterial invasion was usually accompanied by host cell wall dissolution and cellular disruption. Fibrillar materials from degenerated cell walls were usually found in intercellular spaces. Damaged host cells with necrotic cytoplasm showed the localized separation of plasma membrane from the cell wall. Bacterial multiplication and electron-transparent capsule-like structures around bacteria were commonly observed. Meanwhile, cell wall protuberances were prominent outside host cell walls in response to bacterial invasion in Yuzu leaves. Occlusion of intercellular spaces was also formed by the fusion of two or more individual cell wall protuberances originated from two adjacent host cells. Papillae-like materials accumulated locally within host cells in close proximity to bacteria. Some bacteria were found to be undergoing degeneration in xylem vessels. Also, the shrunken, inactive bacteria were surrounded by electron-translucent fibrillar materials in intercellular spaces, implying bacterial immobilization. These cellular responses are thought to be the consequences of defense responses of Yuzu leaves to invading bacteria. In both citrus species, X. citri pv. citri contained polyphosphate bodies showing electron-dense and elliptical structures in cytoplasm. PMID- 19291687 TI - Evaluating antimicrobials and implant materials for infection prevention around transcutaneous osseointegrated implants in a rabbit model. AB - Transcutaneous osseointegrated implants can improve function for select amputee patients, but infection serves as a significant limitation of implantable transcutaneous devices. This study examined the efficacy of an antimicrobial, pexiganan acetate (SUPONEX), and a porous tantalum implant material (Trabecular Metal) in preventing pin tract infection of osseointegrated implants in a rabbit model. Thirty-seven rabbits were randomized to three groups: Ti-control group (n = 11) with titanium alloy implant and no antimicrobial, Ti-Pexiganan group (n = 8) with titanium alloy implant and topical pexiganan acetate 1% applied daily at the skin/implant interface, and Ta-control group (n = 18) with porous tantalum implant and no antimicrobial. All implants were placed transcutaneously through skin, muscle, and bone. Rabbits were monitored for infection for 24 weeks. We observed a 75% reduction in rates of pin tract infection in the Ti-Pexiganan group compared to that observed in the Ti-control group (p = 0.019). No difference in rates of infection was observed between the Ta-control group and the Ti-control group (p = 0.230). In conclusion, pexiganan acetate may be an important antimicrobial for transcutaneous osseointegrated implants. Porous tantalum will not likely prevent pin tract infection without additional methods of soft tissue immobilization around the implant site. PMID- 19291688 TI - A novel hydrophilic poly(lactide-co-glycolide)/lecithin hybrid microspheres sintered scaffold for bone repair. AB - Novel 3-D porous scaffolds made of sintered poly(lacide-co-glycolide) (PLGA)/lecithin hybrid microspheres (PLGA/Lec-SMS) were developed and investigated. The addition of lecithin in PLGA bulk successfully managed the desired hydrophilic modification without sacrificing bulk properties. The outcomes were verified with infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and contact angle analyses. Specifically, this model of scaffold gained significant improvement in mechanical (mainly compressive) strength upon an optimization of lecithin fractions aligning with sintering conditions. Given a perspective of bone tissue engineering use, human fetal osteoblasts were seeded into a series of these PLGA/Lec-SMS scaffolds upon which key parameters of cytocompatibility and osteoconductivity (including cell viability, alkaline phosphatase activity, calcium secretion, and osteogenic genes expression) were assessed. Osteoblasts seeded on PLGA scaffolds with 5 wt % lecithin demonstrated high cell viability and alkaline phosphatase activity. Moreover, elevated lecithin also enhanced the expression of type I collagen. Taken together, these results suggest PLGA/Lec-SMS are promising scaffolds for bone repair. PMID- 19291689 TI - Preparation and evaluation of thiolated chitosan scaffolds for tissue engineering. AB - Thiolated chitosan (TCS) was proposed as a promising candidate as scaffold material for tissue engineering. However, a continuous exploration of such material as a three-dimensional (3D) scaffold with controllable design of microstructure as well as mechanical strength was necessitated. The current study was thus carried out to substantiate such potential of TCS. Thioglycolic acid (TGA) was first introduced on the side chain of chitosan (CS) via the amide bond formation between COOH groups of TGA and NH(2) groups of CS. Composite TCS/CS scaffolds with different ratios were prepared via freeze-drying under different temperatures to optimize the structural properties. The microstructure of the scaffolds was observed by scanning electron microscopy (SEM), and tensile strength of scaffolds was measured. Both the TCS/CS proportion and freezing temperature affected the microstructure and mechanical property of scaffolds, which in turn rendered effects on the growth of cultured fibroblasts. Scaffolds obtained from the TCS/CS proportion of 7:3 and a freezing temperature of -20 degrees C had the maximum tensile strength with a pore distribution ranging from a few to several hundred micrometers. The preferential growth of fibroblasts on this scaffold was also demonstrated. Hence, results in this study would offer valuable information on the preparation of suitable TCS scaffolds for tissue engineering. PMID- 19291690 TI - bFGF-loaded HA-chitosan: a promising scaffold for periodontal tissue engineering. AB - A scaffold containing growth factors promoting regeneration may be a useful device to maintain periodontal regeneration when applied with appropriate cells. The aim of this study is to evaluate the convenience of chitosan and hydroxyapatite (HA)-chitosan scaffolds loaded with basic fibroblast growth factor (bFGF) for periodontal tissue engineering applications. Scaffolds were fabricated by freeze-drying technique using 2 and 3% chitosan gel in the absence or presence of HA particles. Addition of HA beads to chitosan gels produced a novel scaffold in which the pore sizes and interconnectivity were preserved. The scaffolds were loaded with 100 ng bFGF by embedding technique. HA-chitosan scaffolds provide better controlled release kinetics for bFGF compared with chitosan scaffolds and total release continued up to 168 h. Cell culture studies were carried out with periodontal ligament (PDL) cells and cementoblasts. Both 3-[4,5-dimethylthiazol-2 yl]-diphenyltetrazolium bromide (MTT) assay and confocal laser scanning microscope analysis revealed cells proliferating inside the scaffolds. The results demonstrated that bFGF-loaded HA-chitosan scaffolds provide a suitable three-dimensional environment supporting the cellular structure, proliferation, and mineralization. PMID- 19291691 TI - Novel multiarm PEG-based hydrogels for tissue engineering. AB - Injectable scaffolds are promising substrates for regenerative medicine applications. In this study, multiarm amino-terminated poly(ethylene glycol) (PEG) hydrogels were crosslinked with genipin, a compound naturally derived from the gardenia fruit. Four- and eight-arm amino-terminated PEG hydrogels crosslinked with varying concentrations of genipin were characterized. Both surface and cross-sectional structures of PEG-based hydrogels were observed by scanning electron microscopy. In vitro gelation time, water uptake, swelling, and weight loss of PEG hydrogels in phosphate buffered saline at 37 degrees C were studied. The results showed that the eight-arm PEG demonstrated a much slower gelation time compared with the four-arm PEG, which may be due to the differing structures of the multiarm PEG hydrogels, which in turn affects the ability of genipin to react with the amine groups. Human adipose-derived stem cells were seeded onto the four- and eight-arm PEG hydrogels in vitro to assess the biological performance and applicability of the gels as cell carriers. The four arm PEG hydrogel resulted in enhanced cell adhesion when compared with the eight arm PEG hydrogel. Overall, these characteristics provide a potential opportunity for multiarm PEG hydrogels as injectable scaffolds in a variety of tissue engineering applications. PMID- 19291692 TI - Electron impact ion fragmentation pathways of peracetylated C-glycoside ketones derived from cyclic 1,3-diketones. AB - Monosaccharide C-glycoside ketones have been synthesized by aqueous-based Knoevenagel condensation of isotopically labeled and unlabeled aldoses with cyclic diketones, 5,5-dimethyl-1,3-cyclohexanedione (dimedone) and 1,3 cyclohexanedione (1,3-CHD). The reaction products and their corresponding acetylated analogs produce characteristic molecular adduct ions by matrix assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS). Analysis of the peracetylated C-glycosides by electron ionization (EI) gas chromatography/mass spectrometry (GC/MS) revealed diagnostic fragment ions that have been used to deduce the EI fragmentation pathways and the structure of each C-glycoside ketone. Characteristic gluco- and ribo-specific ions were observed at m/z 350 and 278, respectively. Ions common to both carbohydrate fragmentation pathways were observed at m/z 193 and 169 for the dimedone-C-glycosides, and m/z 165 and 141 for the 1,3-CHD-C-glycosides. Ions with m/z 169 and 141 retain the anomeric carbon (carbon-1) of the original sugar, while m/z 193 and 165 are shown to retain carbons-1, 2, and 3. PMID- 19291693 TI - Skin peptides from anurans of the Litoria rubella Group: sequence determination using electrospray mass spectrometry. Opioid activity of two major peptides. AB - Many species of frogs of the genus Litoria secrete bioactive peptides from their skin glands. These peptides are normally host-defence compounds and may have one, or more of the following activities; smooth muscle contraction, analgesic, antimicrobial, antiviral, lymphocyte proliferator (immunomodulator) and neuronal nitric oxide synthase (nNOS) inactivation. Two frog species of the Litoria rubella Group that have been studied before, namely, Litoria electrica and Litoria rubella, are different from other species of the genus Litoria in that they produce small peptides that show neither membrane, lymphocyte nor nNOS activity. In this study we have used electrospray mass spectrometry together with Edman sequencing to identify eight skin peptides of the third member of this Group, Litoria dentata: surprisingly, none of these peptides show activity in our biological screening program. However, two major peptides (FPWL-NH(2) and FPWP NH(2)) from L. electrica and L. rubella are opioids at the micromolar concentration. PMID- 19291694 TI - Sonographic study of the decidua basalis in the first trimester of pregnancy. AB - OBJECTIVES: To describe the sonographic appearance of the decidua basalis and its changes in the first trimester of pregnancy. METHODS: We reviewed images from 159 first-trimester ultrasound examinations in 105 women with uncomplicated pregnancies who later delivered at term. The appearance of the decidua basalis layer and the sonographic changes that it underwent, including in echogenicity and thickness, were analyzed with respect to gestational age. RESULTS: A distinct decidual layer could be identified consistently at 5-6 weeks' gestation and its thickness peaked at 6-7 weeks. It was seen inconsistently at 8-9 weeks and was not identifiable by 10 weeks. Its appearance changed over time, from a uniformly echogenic layer at 5-6 weeks to a heterogeneous echogenic layer at 7 weeks, corresponding to the histological evidence of trophoblast penetration. The layer then became less echogenic with time until it became unidentifiable. CONCLUSIONS: There is a window of opportunity in the first trimester for sonographic examination of the decidua. This may allow screening, at an early stage, for conditions that affect the decidua during pregnancy. PMID- 19291695 TI - Antibiotics: natural products essential to human health. AB - For more than 50 years, natural products have served us well in combating infectious bacteria and fungi. Microbial and plant secondary metabolites helped to double our life span during the 20th century, reduced pain and suffering, and revolutionized medicine. Most antibiotics are either (i) natural products of microorganisms, (ii) semi-synthetically produced from natural products, or (iii) chemically synthesized based on the structure of the natural products. Production of antibiotics began with penicillin in the late 1940s and proceeded with great success until the 1970-1980s when it became harder and harder to discover new and useful products. Furthermore, resistance development in pathogens became a major problem, which is still with us today. In addition, new pathogens are continually emerging and there are still bacteria that are not eliminated by any antibiotic, e.g., Pseudomonas aeruginosa. In addition to these problems, many of the major pharmaceutical companies have abandoned the antibiotic field, leaving much of the discovery efforts to small companies, new companies, and the biotechnology industries. Despite these problems, development of new antibiotics has continued, albeit at a much lower pace than in the last century. We have seen the (i) appearance of newly discovered antibiotics (e.g., candins), (ii) development of old but unutilized antibiotics (e.g., daptomycin), (iii) production of new semi synthetic versions of old antibiotics (e.g., glycylcyclines, streptogrammins), as well as the (iv) very useful application of old but underutilized antibiotics (e.g., teicoplanin). PMID- 19291696 TI - Aerial electrostatic-charged sprays for deposition and efficacy against sweet potato whitefly (Bemisia tabaci) on cotton. AB - BACKGROUND: The efficacy of aerial electrostatic-charged sprays was evaluated for spray deposit characteristics and season-long control of sweet potato whitefly (SWF), Bemisia tabaci Genn. biotype B (aka B. argentifolii Bellows & Perring), in an irrigated 24 ha cotton field. Treatments included electrostatic-charged sprays at full and half active ingredient (AI) label rate, uncharged sprays and conventional sprays applied with CP nozzles at full label rate with several different insecticides. RESULTS: Spray droplet size was significantly smaller for electrostatic-charged sprays than for conventional sprays in top- and mid-canopy locations. The seasonal mean numbers of viable eggs and live large nymphs on cotton treated with electrostatic-charged sprays were comparable with those on cotton treated with conventional applications. Lethal concentration (LC(50)) for adults for electrostatic-charged sprays was comparable with that for conventional sprays. CONCLUSION: The amenability of electrostatic-charged sprays to a wide array of pesticides with different chemistries should be a useful tool in combating insect resistance. Results reported here suggest that the potential exists for obtaining increased efficacy against whiteflies using an electrostatic spray charging system, and that additional research will be required to improve charge-to-mass (Q/M) ratio in order to increase deposition of pest control materials to the lower surfaces of cotton leaves where the whiteflies reside. PMID- 19291697 TI - Leukocyte transmigration is modulated by chemokine-mediated PI3Kgamma-dependent phosphorylation of vimentin. AB - Phosphoinositide 3-kinase gamma (PI3Kgamma) plays a fundamental role in mediating leukocyte migration to inflammation sites. However, the downstream cytoplasmic events triggered by its signaling activity are still largely obscure. To address this issue, tyrosine and serine/threonine phosphorylated proteins of chemokine stimulated WT or PI3Kgamma-null macrophages were investigated. Among the proteins analyzed, the intermediate filament vimentin was found as a downstream effector of the PI3Kgamma signaling pathway. Specific analysis of the phosphorylation state of vimentin in macrophages showed that this protein becomes rapidly phosphorylated in both tyrosine and serine residues upon chemokine stimulation. In the absence of PI3Kgamma or the kinase activity of PI3Kgamma (PI3Kgamma(KD/KD)), phosphorylation of vimentin was reduced. PI3Kgamma-null macrophages displayed impaired chemokine-driven vimentin fiber disassembly as well as reduced ability to transmigrate across endothelial cells. While WT macrophages infected with a vimentin mutant resistant to N-terminal serine phosphorylation showed a reduction in transendothelial migration, infection of PI3Kgamma-null macrophages with a vimentin mutant mimicking serine phosphorylation of N-terminal residues rescued the transendothelial migration defect. These results define vimentin N-terminal phosphorylation and fiber reorganization as a target of chemokine-dependent PI3Kgamma signaling in leukocytes. PMID- 19291698 TI - Negative feedback regulation of IL-32 production by iNOS activation in response to dsRNA or influenza virus infection. AB - iNOS plays an important role in mediating inflammation. In this study, we found that iNOS-derived NO was increased 2.4-fold in the serum samples of 101 patients infected with influenza A virus in comparison with samples of 105 healthy individuals. In A549 human lung epithelial cells, infection with influenza A virus or stimulation with poly(I:C)+IFN-gamma resulted in increased mRNA and protein levels of both IL-32 and iNOS, with subsequent release of NO. Over expression of IL-32 resulted in upregulated iNOS expression with subsequent NO production. Knock down of IL-32 by IL-32-specific siRNA resulted in the inhibition of dsRNA-induced expression of iNOS and NO release, indicating that IL 32 is an upstream regulatory factor of dsRNA-triggered iNOS production. Surprisingly, over-expression of iNOS resulted in the reduction of IL-32 expression, and suppression of iNOS by the selective iNOS inhibitor S methylisothiourea sulfate stimulated IL-32 expression, indicating that a negative feedback mechanism operates between the iNOS/NO and IL-32 systems. These findings suggest that influenza A virus infection activates IL-32 and iNOS expression by a heretofore unrecognized complex mechanism, in which the two pro-inflammatory factors regulate each other, involving positive and negative feedback regulatory loops. PMID- 19291699 TI - Transforming a NEP1 toxin gene into two Fusarium spp. to enhance mycoherbicide activity on Orobanche--failure and success. AB - BACKGROUND: The NEP1 gene encoding a fungal toxin that successfully conferred hypervirulence when transformed into Colletotrichum coccodes (Wallr.) Hughes attacking Abutilon theophrasti (L.) Medic. was tested to ascertain if it would enhance pathogenicity of Fusarium species to Orobanche aegyptiaca Pers. parasitising crops. RESULTS: None of the Fusarium oxysporum (#CNCM I-1622) NEP1 transformants was hypervirulent. NEP1 transformants of a new but unnamed Fusarium sp. (#CNCM I-1621--previously identified as F. arthrosporioides) killed Orobanche more rapidly than the wild type. Transformed lines of both species were NEP1 PCR positive, as was the wild type of F. oxysporum #CNCM I-1622 and five other formae speciales of F. oxysporum. All six wild-type formae speciales of F. oxysporum tested excrete minute amounts of immunologically and bioassay-detectable NEP1 like protein. NEP1 expression of most F. oxysporum transformants was suppressed, suggesting that the native gene and the transgene silence each other. The sequence of the putative NEP1 gene in Fusarium oxysporum #CNCM I-1622 differs from the sequence in the toxin-overproducing strain of F. oxysporum f. sp. erythroxyli in four or five amino acids in the first exon. CONCLUSION: Wild-type Fusarium sp. #CNCM I-1621 does not contain a NEP1-like gene, explaining why it seemed amenable to transformation with high expression, and its virulence was probably enhanced by not cosuppressing the endogenous gene as occurred with Fusarium oxysporum #CNCM I-1622. PMID- 19291700 TI - Synergy-inducing chemokines enhance CCR2 ligand activities on monocytes. AB - The migration of monocytes to sites of inflammation is largely determined by their response to chemokines. Although the chemokine specificities and expression patterns of chemokine receptors are well defined, it is still a matter of debate how cells integrate the messages provided by different chemokines that are concomitantly produced in physiological or pathological situations in vivo. We present evidence for one regulatory mechanism of human monocyte trafficking. Monocytes can integrate stimuli provided by inflammatory chemokines in the presence of homeostatic chemokines. In particular, migration and cell responses could occur at much lower concentrations of the CCR2 agonists, in the presence of chemokines (CCL19 and CCL21) that per se do not act on monocytes. Binding studies on CCR2(+) cells showed that CCL19 and CCL21 do not compete with the CCR2 agonist CCL2. Furthermore, the presence of CCL19 or CCL21 could influence the degradation of CCL2 and CCL7 on cells expressing the decoy receptor D6. These findings disclose a new scenario to further comprehend the complexity of chemokine-based monocyte trafficking in a vast variety of human inflammatory disorders. PMID- 19291701 TI - Natural Treg cells spontaneously differentiate into pathogenic helper cells in lymphopenic conditions. AB - Induction of Forkhead-box p3 (Foxp3) expression in developing T cells upon peptide-MHC encountering has been proposed to define a lineage of committed Treg cells. However, sustained expression of Foxp3 is required for Treg function and what maintains Foxp3 expression in peripheral Treg remains obscure. To address this issue, we monitored natural Treg phenotype and function upon adoptive transfer into lymphocyte-deficient mice. We first show that about 50% of Foxp3 GFP(+) Treg isolated from Foxp3(gfp) KI animals loose Foxp3 expression in severe lymphopenic conditions. We next evidence that the cytokine IL-2, either produced by co-transferred conventional T cells or administrated i.v. prevents Foxp3 downregulation. Moreover, we document that Treg that lost Foxp3 expression upon adoptive transfer produce IL-2 are not suppressive and promote tissue infiltration and damage upon secondary transfer into alymphoid mice. Our findings that Treg convert into pathogenic Th cells in absence of IL-2 provide new clues to the success of Treg-based immune therapies. PMID- 19291702 TI - Human self-protein CD8+ T-cell epitopes are both positively and negatively selected. AB - The cellular immune system recognizes self-epitopes in the context of MHC-I molecules. The immunological general view presumes that these self-epitopes are just a background, both positively and negatively selecting T cells. We here estimate the number of epitopes in each human protein for many frequent HLA alleles, and a score representing over or under presentation of epitopes on these proteins. We further show that there is a clear selection for the presentation of specific self-protein types. Proteins presenting many epitopes include, for example, autoimmune regulator (AIRE) upregulated tissue-specific antigens, immune system receptors and proteins with a high expression level. On the other hand, proteins that may be considered less "useful" for the immune system, such as low expression level proteins, are under-presented. We combine our epitope estimate with single nucleotide polymorphism (SNP) measures to show that this selection can be directly observed through the fraction of non-synonymous SNP (replacement fraction), which is significantly higher inside epitopes than outside. PMID- 19291703 TI - Reciprocal regulation of IL-23 and IL-12 following co-activation of Dectin-1 and TLR signaling pathways. AB - Recognition of microbial products by germ-line-encoded PRR initiates immune responses, but how PRR mediate specific host responses to infectious agents is poorly understood. We and others have proposed that specificity is achieved by collaborative responses mediated between different PRR. One such example comprises the fungal beta-glucan receptor Dectin-1, which collaborates with TLR to induce TNF production. We show here that collaborative responses mediated by Dectin-1 and TLR2 are more extensive than first appreciated, and result in enhanced IL-23, IL-6 and IL-10 production in DC, while down-regulating IL-12 relative to the levels produced by TLR ligation alone. Such down-regulation occurred with multiple MyD88-coupled TLR, was dependent on signaling through Dectin-1 and also occurred in macrophages. These findings explain how fungi can induce IL-23 and IL-6, while suppressing IL-12, a combination which has previously been shown to contribute to the development of Th17 responses found during fungal infections. Furthermore, these data reveal how the collaboration of different PRR can tailor specific responses to infectious agents. PMID- 19291704 TI - Schistosoma mansoni egg antigens induce Treg that participate in diabetes prevention in NOD mice. AB - Schistosoma mansoni soluble egg antigens (SEA) profoundly regulate the infected host's immune system. We previously showed that SEA prevents type 1 diabetes in NOD mice and that splenocytes from SEA-treated mice have reduced ability to transfer diabetes to NOD.scid recipients. To further characterize the mechanism of diabetes prevention we examined the cell types involved and showed that CD25(+) T-cell depletion of splenocytes from SEA-treated donors restored their ability to transfer diabetes. Furthermore, SEA treatment increased the number and proportional representation of Foxp3(+) T cells in the pancreas of NOD mice. We have used in vitro systems to analyze the effect of SEA on the development of NOD Foxp3(+) T cells. We find that SEA can induce Foxp3 expression in naive T cells in a TGF-beta-dependent manner. Foxp3 induction requires the presence of DC, which we also show are modified by SEA to upregulate C-type lectins, IL-10 and IL 2. Our studies show that SEA can have a direct effect on CD4(+) T cells increasing expression of TGF-beta, integrin beta8 and galectins. These effects of SEA on DC and T cells may act in synergy to induce Foxp3(+) Treg in the NOD mouse. PMID- 19291705 TI - The influence of simulated immigration and chemical persistence on recovery of macroinvertebrates from cypermethrin and 3,4-dichloroaniline exposure in aquatic microcosms. AB - BACKGROUND: Chemical dissipation and organism immigration are considered important factors that influence recovery potential from perturbation of aquatic macroinvertebrates. Experiments were conducted to investigate the effect of simulated immigration on recovery of aquatic macroinvertebrates exposed in outdoor microcosms to ecotoxicologically similar concentrations of the rapidly dissipating pyrethroid insecticide cypermethrin (70 ng L(-1)) or the more persistent herbicide intermediate and degradate 3,4-dichloroaniline (10 mg L( 1)). Microcosms were covered with light-permeable mesh to prevent recolonisation. Immigration was simulated by the regular addition of organisms after treatment. RESULTS: Microcosms exposed to 3,4-dichloroaniline treatment suffered substantial loss of taxon richness and by 10 months after treatment had only recovered where invertebrates had been added. Those treated with cypermethrin underwent an initial decline in certain crustacean and insect populations. These populations showed some signs of recovery over a period of 5 months through internal processes alone. However, rate of recovery was further enhanced where immigration was simulated, and in this case recovery had occurred around 100 days after treatment. CONCLUSION: Although not the only factors involved, simulated immigration and chemical fate clearly influence the ability of communities to recover from chemical exposure. Consideration of immigration processes and development of models will help to increase the realism of risk assessments. PMID- 19291706 TI - A thermostable triple mutant of pyranose 2-oxidase from Trametes multicolor with improved properties for biotechnological applications. AB - In order to increase the thermal stability and the catalytic properties of pyranose oxidase (P2Ox) from Trametes multicolor toward its poor substrate D galactose and the alternative electron acceptor 1,4-benzoquinone (1,4-BQ), we designed the triple-mutant T169G/E542K/V546C. Whereas the wild-type enzyme clearly favors D-glucose as its substrate over D-galactose [substrate selectivity (k(cat)/K(M))(Glc)/(k(cat)/K(M))(Gal) = 172], the variant oxidizes both sugars equally well [(k(cat)/K(M))(Glc)/(k(cat)/K(M))(Gal) = 0.69], which is of interest for food biotechnology. Furthermore, the variant showed lower K(M) values and approximately ten-fold higher k(cat) values for 1,4-BQ when D-galactose was used as the saturating sugar substrate, which makes this enzyme particularly attractive for use in biofuel cells and enzyme-based biosensors. In addition to the altered substrate specificity and reactivity, this mutant also shows significantly improved thermal stability. The half life time at 60 degrees C was approximately 10 h, compared to 7.6 min for the wild-type enzyme. We performed successfully small-scale bioreactor pilot conversion experiments of D-glucose/D galactose mixtures at both 30 and 50 degrees C, showing the usefulness of this P2Ox variant in biocatalysis as well as the enhanced thermal stability of the enzyme. Moreover, we determined the crystal structure of the mutant in its unligated form at 1.55 A resolution. Modeling D-galactose in position for oxidation at C2 into the mutant active site shows that substituting Thr for Gly at position 169 favorably accommodates the axial C4 hydroxyl group that would otherwise clash with Thr169 in the wild-type. PMID- 19291707 TI - A microreactor for the study of biotransformations by a cross-linked gamma lactamase enzyme. AB - A (+)-gamma-lactamase was precipitated, cross-linked and the resulting solid crushed prior to immobilisation within a capillary column microreactor. The microreactor was subsequently used to study enzyme stability, activity, kinetics and substrate specificity. The thermophilic (+)-gamma-lactamase retained 100% of its initial activity at the assay temperature, 80 degrees C, for 6 h and retained 52% activity after 10 h, indicating the advantage of immobilisation. This high stability of the immobilised enzyme provided the advantage that it could be utilised to screen many compounds in the microreactor system. This advantage overcame the fact that the immobilisation process affected enzyme kinetics and activity, which was reduced (by 70%) compared to the free enzyme. In general, the enzyme displayed similar substrate specificity to that found in a previous study for the free enzyme; however, enhanced activity was seen towards one substrate, acrylamide. The system developed correlates well with the free enzyme in batch assay and indicates the suitability of the system for enzyme substrate screening, allowing a significant reduction in cost, due to the reduced amounts of enzyme, substrates and other assay constituents required. PMID- 19291709 TI - Do we see what we think we see? The complexities of morphological assessment. AB - Reliable pathological interpretation is vital to so many aspects of tissue-based research as well as being central to patient care. Understanding the complex processes involved in decision-making is the starting point to improve both diagnostic reproducibility and the definition of diagnostic groups that underpin our experiments. Unfortunately, there is a paucity of research in this field and it is encouraging to see The Journal of Pathology publishing work in this area. This review attempts to highlight the opportunities that exist in this field and the technologies that are now available to support this type of research. Key amongst these are the use of decision analysis tools such as inference networks, and virtual microscopy that allows us to simulate diagnostic decision-making. These tools have roles, not only in studying the subtleties of diagnostic decision-making, but also in delivering new methods of training and proficiency testing. Research which helps us to better understand what we see, why we see it, and standardizing interpretative reasoning in pathological classification is essential for improving the wide range of activities that pathologists support, including clinical diagnosis, teaching, training, and experimental research. PMID- 19291710 TI - Acute hypoxia and osteoclast activity: a balance between enhanced resorption and increased apoptosis. AB - Osteoclasts are the primary mediators of pathological bone resorption in many conditions in which micro-environmental hypoxia is associated with disease progression. However, effects of hypoxia on human osteoclast activity have not been reported. Mature human osteoclasts were differentiated from peripheral blood or obtained from giant cell tumour of bone. Osteoclasts were exposed to a constant hypoxic environment and then assessed for parameters including resorption (toluidine blue staining of dentine slices), membrane integrity (trypan blue exclusion), apoptosis (TUNEL, DAPI), and osteolysis-associated enzyme activity (TRAP, cathepsin K). 24 h exposure to 2% O(2) produced a 2.5-fold increase in resorption associated with increased TRAP and cathepsin K enzyme activity. Hypoxia-Inducible Factor-1alpha (HIF-1alpha) siRNA completely ablated the hypoxic increase in osteoclast resorption. 24 h at 2% O(2) also increased the number of osteoclasts with compromised membrane integrity from 6% to 21%, with no change in the total osteoclast number or the proportion of late-stage apoptotic cells. Transient reoxygenation returned the percentage of trypan blue-positive cells to normoxic levels, suggesting that osteoclasts can recover from the early stages of cell death. Repeated over an extended period, hypoxia/reoxygenation enhanced osteoclast differentiation at this pO(2). These data suggest that in diseased bone, where the pO(2) may fall to 50 mM, respectively. At elevated reaction temperatures, we obtained Pt nanonetworks and Pt nanodendrites in the presence of SDS at concentrations of <10 mM and >50 mM, respectively. Transmission electron microscopy images revealed that these Pt NMs were all composed of one dimensional Pt nanostructures having widths of 3.0+/-1.0 nm and lengths of 17.0+/-4.8 nm. Cyclic voltammetry data indicated that the as-prepared Pt nanonetworks, nanosponges, and nanodendrites possessed large electrochemically active surface areas (77.0, 70.4, and 41.4 m(2) g(-1), respectively). For the electrocatalytic oxidation of methanol, the ratio of the forward oxidation peak current (I(f)) to the backward peak current (I(b)) of the Pt nanodendrites, nanosponges, and nanonetworks were all high (I(f)/I(b)=2.88, 2.66, and 2.16, respectively). These three NMs exhibit greater electrocatalytic activities and excellent tolerance toward poisoning species for the oxidation of methanol when compared with the performance of standard Pt NMs. PMID- 19291725 TI - Unprecedented solvent-assisted reactivity of Hydrido W3CuS4 cubane clusters: the non-innocent behaviour of the cluster-core unit. AB - Opening the cluster core: Substitution of the chloride ligand in the novel cationic cluster [W(3)CuS(4)H(3)Cl(dmpe)(3)](+) (see figure; dmpe=1,2 bis(dimethylphosphino)ethane) by acetonitrile is promoted by water addition. Kinetic and density functional theory studies lead to a mechanistic proposal in which acetonitrile or water attack causes the opening of the cluster core with dissociation of one of the Cu--S bonds to accommodate the entering ligand.Reaction of the incomplete cuboidal cationic cluster [W(3)S(4)H(3)(dmpe)(3)](+) (dmpe=1,2-bis(dimethylphosphino)ethane) with Cu(I) compounds produces rare examples of cationic heterodimetallic hydrido clusters of formula [W(3)CuClS(4)H(3)(dmpe)(3)](+) ([1](+)) and [W(3)Cu(CH(3)CN)S(4)H(3)(dmpe)(3)](2+) ([2](2+)). An unexpected conversion of [1](+) into [2](2+), which involves substitution of chloride by CH(3)CN at the copper centre, has been observed in CH(3)CN/H(2)O mixtures. Surprisingly, formation of the acetonitrile complex does not occur in neat acetonitrile and requires the presence of water. The kinetics of this reaction has been studied and the results indicate that the process is accelerated when the water concentration increases and is retarded in the presence of added chloride. Computational studies have also been carried out and a mechanism for the substitution reaction is proposed in which attack at the copper centre by acetonitrile or water causes disruption of the cubane-type core. ESI-MS experiments support the formation of intermediates with an open-core cluster structure. This kind of process is unprecedented in the chemistry of M(3)M'Q(4) (M=Mo, W; Q=S, Se) clusters, and allows for the transient appearance of a new coordination site at the M' site which could explain some aspects of the reactivity and catalytic properties of this kind of clusters. PMID- 19291726 TI - Electronic energy transfer to the S2 level of the acceptor in functionalised boron dipyrromethene dyes. AB - Taking the high road: Highly efficient electronic energy transfer takes place from a set of appended aryl polycyclic hydrocarbons to an expanded boron dipyrromethene (Bodipy)-based dye (see figure) despite negligible spectral overlap with the lowest-energy excited state localised on the acceptor.A multi component array has been constructed around an expanded boron dipyrromethene (Bodipy) dye that absorbs and emits in the far-red region. One of the appendages is a perylene-based moiety that is connected to the boron atom of the terminal Bodipy by a 1,4-diethynylphenylene connector. Despite the fact that there is almost negligible spectral overlap between fluorescence from the perylene unit and absorption by the Bodipy residue, electronic energy transfer is rapid and essentially quantitative. It is concluded that at least half of the photons absorbed by perylene are transferred to the upper-lying singlet excited state (S(2)) associated with the Bodipy-based acceptor. The second appendage is a pyrene unit that is covalently linked to fluorene, through an ethynylene spacer, and to the boron atom of the Bodipy terminus, through a 1,4-diethynylphenylene connector. Pyrene absorbs and emits at higher energy than perylene and there is strong spectral overlap with the Bodipy-based S(2) state, and none with the corresponding S(1) state. Electronic energy transfer is now very fast and exclusively to the S(2) state of the acceptor. It is difficult to compute reasonable estimates for the rates of Coulombic energy transfer, because of uncertainties in the orientation factor, but the principle mechanism is believed to arise from electron exchange. Comparison with an earlier array built around a conventional Bodipy dye indicates that there are comparable electronic coupling matrix elements for the two systems. It is notable that pyrene is more strongly coupled to the Bodipy unit than perylene in both arrays. These new arrays function as highly effective solar concentrators. PMID- 19291727 TI - Urinary cysteinyl leukotriene E4 significantly increases during pain in children and adults with sickle cell disease. AB - Baseline level of the cysteinyl leukotriene (CysLT), leukotriene E4 (LTE4), is associated with an increased pain rate in children and adults with sickle cell disease (SCD). To provide additional evidence for a role of CysLTs in the pathogenesis of vaso-occlusion, we tested the hypothesis that LTE4 levels will increase within an individual during painful episodes compared to baseline. In a cohort of 19 children and adults with SCD, median LTE4 levels increased from 82.36 pg/mg creatinine at baseline to 162.81 pg/mg creatinine during a painful episode (P < 0.001). These data further support a contribution of CysLTs to the process of vaso-occlusion. PMID- 19291728 TI - Enzyme, substrate, and myeloma in Gaucher disease. PMID- 19291729 TI - Safety of anti-D during pregnancy. PMID- 19291730 TI - Lyotropic liquid-crystalline solutions of high-concentration dispersions of single-walled carbon nanotubes with conjugated polymers. PMID- 19291731 TI - Destruction and control of Toxoplasma gondii tachyzoites using gold nanosphere/antibody conjugates. PMID- 19291732 TI - 3D metallic nanostructure fabrication by surfactant-assisted multiphoton-induced reduction. PMID- 19291733 TI - Optical tweezers study of topoisomerase inhibition. PMID- 19291734 TI - Robust Au-PEG/PS microbeads as optically stable platforms for SERS. PMID- 19291735 TI - Preparation of metallic line patterns from functional block copolymers. AB - A simple route for the preparation of nanoscopic metallic line patterns from functional block copolymers (BCPs) containing poly(2-vinylpyridine) or poly(methyl methacrylate) blocks is demonstrated. The time evolution of the surface morphologies of BCP thin films exposed to solvent vapors is studied to optimize the conditions for generating BCP microdomains oriented parallel or normal to the substrate. BCP templates are prepared by film reconstruction or by removal of one of the copolymer microdomains, depending on the properties of the functional BCPs. Finally, metallic line patterns are prepared by either electrochemical etching or direct metal deposition and lift-off processes using the BCP templates. PMID- 19291736 TI - Synthesis, modelling, and antimitotic properties of tricyclic systems characterised by a 2-(5-Phenyl-1H-pyrrol-3-yl)-1,3,4-oxadiazole moiety. AB - Antitumour activity was observed in a series of tricyclic compounds characterised by a 2-(1H-pyrrol-3-yl)-1,3,4-oxadiazole moiety with various substitutions. Their synthesis and antiproliferative activity toward a panel of human tumour cell lines is described. The most interesting compounds 1 c and 4 c were selected for further evaluation to elucidate their possible mechanism of action.Interesting antitumour activity was observed in a series of tricyclic compounds characterised by the presence of a 2-(1H-pyrrol-3-yl)-1,3,4-oxadiazole moiety that is variously substituted. Their synthesis and antiproliferative activity toward a panel of human tumour cell lines is described. The two most interesting compounds were selected for further evaluation to elucidate their possible mechanism of action. Analysis of cell cycle, tubulin polymerisation, modulation of mitotic markers of the M phase, and apoptosis showed that antimitotic activity is the primary mechanism of the cytotoxic effects of these compounds. Experiments performed on isolated tubulin confirmed that the compounds act by inducing tubulin polymerisation, like taxanes. The binding model against tubulin was also examined by molecular modelling and docking. The results support the proposed binding model, which is able to explain the activity of the oxadiazole derivatives on the basis of their docking energy. PMID- 19291737 TI - Proteomic analysis for process development and control of therapeutic protein separation from human plasma. AB - The use of proteomics technology during the development of a new process for plasma protein separation was demonstrated. In a two-step process, the two most abundant proteins, HSA and IgG, were removed in a first step of anion-exchange chromatography using a gel with very high capacity. Subsequently, two fractions containing medium and low abundance proteins were re-chromatographed on a smaller column with the same type of gel. Collected fractions were separated by SDS-PAGE and 2-D electrophoresis, and excised proteins were digested with trypsin and identified by LC-ESI-MS/MS. This proteomic analysis proved to be a useful method for detection of low abundance therapeutic proteins and potential harmful contaminants during process development. Based on this method, low abundance therapeutic proteins, such as vitamin-K-dependent clotting factors and inhibitors, could be identified as present in target fractions after chromatographic separation. In addition, the tracking of potentially dangerous impurities and designing proper steps for their removal are important outcomes when developing, refining or controlling a new fractionation schema. For the purpose of in-process control, in-solution digestion of complete fractions followed by protein identification with LC-ESI-MS/MS was demonstrated as a rapid and simple alternative to the entire analysis including 1-D or 2-D electrophoretic steps. PMID- 19291738 TI - Multiple solvent crystal structures of ribonuclease A: an assessment of the method. AB - The multiple solvent crystal structures (MSCS) method uses organic solvents to map the surfaces of proteins. It identifies binding sites and allows for a more thorough examination of protein plasticity and hydration than could be achieved by a single structure. The crystal structures of bovine pancreatic ribonuclease A (RNAse A) soaked in the following organic solvents are presented: 50% dioxane, 50% dimethylformamide, 70% dimethylsulfoxide, 70% 1,6-hexanediol, 70% isopropanol, 50% R,S,R-bisfuran alcohol, 70% t-butanol, 50% trifluoroethanol, or 1.0M trimethylamine-N-oxide. This set of structures is compared with four sets of crystal structures of RNAse A from the protein data bank (PDB) and with the solution NMR structure to assess the validity of previously untested assumptions associated with MSCS analysis. Plasticity from MSCS is the same as from PDB structures obtained in the same crystal form and deviates only at crystal contacts when compared to structures from a diverse set of crystal environments. Furthermore, there is a good correlation between plasticity as observed by MSCS and the dynamic regions seen by NMR. Conserved water binding sites are identified by MSCS to be those that are conserved in the sets of structures taken from the PDB. Comparison of the MSCS structures with inhibitor-bound crystal structures of RNAse A reveals that the organic solvent molecules identify key interactions made by inhibitor molecules, highlighting ligand binding hot-spots in the active site. The present work firmly establishes the relevance of information obtained by MSCS. PMID- 19291739 TI - Solution structure of the Escherichia coli HybE reveals a novel fold. PMID- 19291740 TI - NMR-derived solution structure of SUMO from Drosophila melanogaster (dSmt3). PMID- 19291741 TI - A coarse-grained potential for fold recognition and molecular dynamics simulations of proteins. AB - A coarse-grained potential for protein simulations and fold ranking is presented. The potential is based on a two-point model of individual amino acids and a specific implementation of hydrogen bonding. Parameters are determined for distance dependent pair interactions, pseudo bonds, angles, and torsions. A scaling factor for a hydrogen bonding term is also determined. Iterative sampling for 4867 proteins reproduces distributions of internal coordinates and distances observed in the Protein Data Bank. The adjustment of the potential and resampling are in the spirit of the generalized ensemble approach. No native structure information (e.g., secondary structure) is used in the calculation of the potential or in the simulation of a particular protein. The potential is subject to two tests as follows: (i) simulations of 956 globular proteins in the neighborhood of their native folds (these proteins were not used in the training set) and (ii) discrimination between native and decoy structures for 2470 proteins with 305,000 decoys and the "Decoys 'R' Us" dataset. In the first test, 58% of tested proteins stay within 5 A from the native fold in Molecular Dynamics simulations of more than 20 nanoseconds using the new potential. The potential is also useful in differentiating between correct and approximate folds providing significant signal for structure prediction algorithms. Sampling with the potential consistently regenerates the distribution of distances and internal coordinates it learned. Nevertheless, during Molecular Dynamics simulations structures are found that reproduce the learned distributions but are far from the native fold. PMID- 19291742 TI - Effects of chlorantraniliprole on eggs and larvae of Lobesia botrana (Denis & Schiffermuller) (Lepidoptera: Tortricidae). AB - BACKGROUND: Chlorantraniliprole is a novel insecticide belonging to the class of selective ryanodine receptor agonists. The aims of this study were to evaluate the toxicity and the length of residual activity of chlorantraniliprole against the grapevine moth, Lobesia botrana (Denis & Schiffermuller), larvae and eggs, under laboratory and field conditions. RESULTS: Dose-response bioassays showed that chlorantraniliprole had a high level of toxicity to neonate larvae of L. botrana. In field trials, chlorantraniliprole applied at 35 mg AI L(-1) was as effective against the larvae as the commercial standard reference, chlorpyrifos ethyl, and had a long-lasting effect. When applied at a field rate of 35 mg AI L( 1), chlorantraniliprole caused greater than 20% egg mortality. The toxic effect of chlorantraniliprole is completed by its substantial activity against neonates emerging from eggs. CONCLUSION: Chlorantraniliprole can be considered as a useful tool for L. botrana control, providing a new standard in efficacy and an extremely good residual control. The combined effects against eggs and larvae of grapevine moth should contribute to its efficacy in the field. The new mode of action of chlorantraniliprole and its safety to beneficials and non-target organisms would be extremely useful in IPM and for managing resistance to insecticides. PMID- 19291743 TI - Assessing the efficiency of mixed effects modelling in quantifying metabolism based drug-drug interactions: using in vitro data as an aid to assess study power. AB - The clinical assessment of metabolic drug-drug interactions (mDDI) may involve population-based pharmacokinetic (POPPK) assessment as part of Phase 3 clinical trials. The elements of such POPPK study design have not been linked to prior information from in vitro experiments. Using in vitro-in vivo extrapolation techniques, implemented within Simcyp algorithms, the influence of POPPK study design (sample size, concentration-time data points, proportion of the population receiving a concomitant medication (COMED)) was studied in relation to the inhibitory potency of the each COMED. Steady-state concentrations of a candidate compound (compound X; mainly metabolized by cytochrome P450 enzymes, CYP3A4 and CYP2D6) in the presence and absence of COMEDs (including ketoconazole, fluconazole, quinidine and paroxetine as inhibitors) were analysed using non linear mixed effect modelling (NONMEM). The NONMEM operator was blind to the nature of the COMEDs and the inhibitory effects on model parameters were classified as either statistically (p>0.01 for a change in objective function) or kinetically (COMED effect>2 fold) significant. Using a population study size of 2000, no false-positive cases were identified and, except in one case, no false negative interaction was observed when >2.5% of patients had received an interacting COMED. The findings increase the confidence in the ability of the mixed effects modelling approach to identify 'true' interactions. However, they also emphasize the importance of study design and the potential value of using pre-clinical information from in vitro studies. Recent US Food and Drug Administration guidance on mDDI has put more emphasis on the use of in vitro systems for detecting and anticipating such effects. Combining these data with the framework of non-linear mixed effect modelling seems a natural progression in the field of assessing mDDI. PMID- 19291744 TI - Male-male combats in a polymorphic lizard: residency and size, but not color, affect fighting rules and contest outcome. AB - Theoretical models predict that the outcome of dyadic agonistic encounters between males is influenced by resource-holding potential, resource value, and intrinsic aggressiveness of contestants. Moreover, in territorial disputes residents enjoy a further obvious competitive advantage from the residency itself, owing to the intimate familiarity with their territory. Costs of physical combats are, however, dramatically high in many instances. Thus, signals reliably reflecting fighting ability of the opponents could easily evolve in order to reduce these costs. For example, variation in color morph in polymorphic species has been associated with dominance in several case studies. In this study, we staged asymmetric resident-intruder encounters in males of the common wall lizard Podarcis muralis, a species showing three discrete morphs (white, yellow, and red) to investigate the effects of asymmetries in color morph, residency, and size between contestants on the outcome of territorial contests. We collected aggression data by presenting each resident male with three intruders of different color morph, in three consecutive tests conducted in different days, and videotaping their interactions. The results showed that simple rules such as residency and body size differences could determine the outcome of agonistic interactions: residents were more aggressive than intruders, and larger males were competitively superior to smaller males. However, we did not find any effect of color on male aggression or fighting success, suggesting that color polymorphism in this species is not a signal of status or fighting ability in intermale conflicts. PMID- 19291745 TI - Complications of fractional CO2 laser resurfacing: four cases. AB - BACKGROUND AND OBJECTIVE: Fractional ablative laser therapy is a new modality which will likely be widely used due to its efficacy and limited side-effect profile. It is critical to recognize, characterize, and report complications in order to acknowledge the limits of therapeutic efficacy and to improve the safety of these devices. STUDY DESIGN/MATERIALS AND METHODS: The photographs, treatment parameters, and clinical files of four female patients aged 54-67 who had scarring or ectropion after fractional CO(2) laser resurfacing on the face or neck were carefully reviewed to search for any possible linking factors. RESULTS: Patient 1 developed erosions and swelling of the right lower eyelid 2 days postoperatively, which developed into scarring and an ectropion. Patient 2 developed linear erosions and beefy red swelling on the right side of the neck which developed into a tender, band-like scar over 1-month. Patient 3 developed stinging and yellow exudate in multiple areas of the neck 3 days postoperatively. Cultures grew methicillin-resistant Staphylococcus aureus. Despite appropriate treatment, she developed multiple areas of irregular texture and linear streaking which developed into scars. Patient 4 developed an asymptomatic patchy, soft eschar with yellowish change on the left side of the neck. Azithromycin was started, however at 2-week follow-up she had fibrotic streaking which developed into horizontal scars and a vertical platysmal band. The treatment and final outcome of each patient are described. CONCLUSION: Scarring after fractional CO(2) laser therapy may be due to overly aggressive treatments in sensitive areas (including excessive energy, density, or both), lack of technical finesse, associated infection, or idiopathic. Care should be taken when treating sensitive areas such as the eyelids, upper neck, and especially the lower neck and chest by using lower energy and density. Postoperative infections may lead to scarring and may be prevented by careful taking of history, vigilant postoperative monitoring and/or prophylactic antibiotics. PMID- 19291746 TI - Hypertrophic scarring of the neck following ablative fractional carbon dioxide laser resurfacing. AB - BACKGROUND: Ablative fractional carbon dioxide (CO(2)) laser treatments have gained popularity due to their efficacy, shortened downtime, and decreased potential for scarring in comparison to traditional ablative CO(2) resurfacing. To date, scarring with fractional CO(2) lasers has not been reported. OBJECTIVE: Five patients treated with the same fractional CO(2) laser technology for photodamage of the neck were referred to our practices 1-3 months after treatment. Each patient developed scarring. Of the five cases, two are discussed in detail. The first was treated under general anesthesia on the face and anterior neck at a pulse energy of 30 mJ (859 microm depth) with 25% coverage. Eleven days after treatment, three non-healing areas along the horizontal skin folds of the anterior neck were noted. At 2 weeks after CO(2) ablative fractional resurfacing, these areas had become thickened. These raised areas were treated with a non-ablative fractionated 1,550 nm laser to modify the wound healing milieu. One week later, distinct firm pale papules in linear arrays with mild hypopigmentation had developed along involved neck skin folds. Skin biopsy was performed. For the second patient, the neck was treated at a pulse energy of 20 mJ (630 microm depth) with 30% coverage of the exposed skin, with a total treatment energy of 5.0 kJ. Minimal crusting was noted on the neck throughout the initial healing phase of 2 weeks. She then experienced tightness on her neck. Approximately 3 weeks after treatment, she developed multiple vertical and horizontal hypertrophic scars (HS). RESULTS: Histopathology for the first case confirmed the presence of a hypertrophic scar. The papules in this case completely resolved with mild residual hypopigmentation after treatment with topical corticosteroids. HS failed to resolve in the second case to date after 1 month. CONCLUSION: As with traditional ablative CO(2) laser resurfacing, HS is a potential complication of ablative fractional CO(2) laser resurfacing, particularly on the neck. With early diagnosis and appropriate treatment HS of neck skin may be reversible. We urge caution when treating the neck with this device and close attention to wound care in the post-operative period. PMID- 19291747 TI - Fractional nonablative 1540 nm laser resurfacing for thermal burn scars: a randomized controlled trial. AB - BACKGROUND AND OBJECTIVE: Burn scars cause permanent and disfiguring problems for many patients and limited treatments are available. Nonablative fractional lasers induce a wound healing response, which may lead to remodeling of burn scar texture. This randomized trial evaluates efficacy and adverse effects of 1540 nm fractional laser versus untreated control for burn scars. MATERIALS AND METHODS: Seventeen adult patients with burn scars of 1 year or older and Fitzpatrick skin types I-III were included in the study. Side-by-side test areas were randomized to (i) three monthly 1540 nm fractional laser treatments and (ii) no treatment. Blinded on-site response evaluations were performed 4 and 12 weeks after final treatment. Primary endpoints were change in skin texture (0-10 categorical scale), pain and adverse effects. Secondary endpoints were change in skin color and patient satisfaction (0-10 categorical scales). RESULTS: Preoperative skin texture was similar and moderately uneven in treated and untreated sites (6 (5.5 8), P = 1). Postoperative laser-treated skin appeared significantly more even and smooth compared to adjacent untreated control sides (4 weeks: 4 (2.5-6), P = 0.0015; 12 weeks: 4 (2-5), P = 0.0007). Patients were satisfied with treatments (week 12: 7 (4-8.5)) and 8/17 patients evaluated burn scars to be moderately or significantly improved. Skin redness increased transiently from laser treatments. No significant differences were found in skin pigmentation. Patients experienced moderate pain (5 (3.5-6)), erythema (17/17/16 patients, after first/second/third treatments), edema (9/9/8), bullae (3/0/3), and crusts (4/0/4). One patient had minor scarring. No adverse effects were seen in untreated control areas. CONCLUSIONS: Nonablative 1540 nm fractional laser improves burn scar texture, which raises a new potential for future burn scar treatment. PMID- 19291748 TI - Fractional ablative skin resurfacing: complications. PMID- 19291749 TI - Pulsing influences photoradiation outcomes in cell culture. AB - BACKGROUND AND OBJECTIVES: Skin pigmentation can adversely affect phototherapy outcomes. Delivering pulsed light has been suggested as a means of enhancing efficacy. Suitable pulse frequencies remain indeterminate, often being selected empirically. This study was undertaken to determine whether pulsed light delivery mitigates the filtering effect of melanin pigment on photomodulation in vitro. STUDY DESIGN AND METHODS: Human HEP-2 cells were cultured in complete DMEM media. Photoradiation was delivered through 0.025% melanin filters at 670 nm (5.0 J/cm(2)/treatment/24 hours) for 72 hours at different pulse rates. Group A received no light treatment. Group B received treatments without pulsing. Groups C, D, E, F, and G received treatments at 6, 18, 36, 100, and 600 Hz. Cell proliferation was assessed by MTT assay and oxidative burst was measured using the 2.7 dichloro-fluorescein-diacetate assay. RESULTS: Cell proliferation was maximally stimulated at 100 Hz at 48 and 72 hours (n = 4, P< or =0.05). Oxidative burst was maximally stimulated at 600 Hz (n = 4, P< or =0.05). All frequencies were stimulatory at 48 and 72 hours (n = 4, P< or =0.05). CONCLUSION: This investigation suggests that light pulsing may improve outcomes by mitigating the filtration effects of cutaneous melanin. Further studies to further define these effects are warranted. PMID- 19291750 TI - Risk factors for recurrence of vaginal intraepithelial neoplasia in the vaginal vault after laser vaporization. AB - BACKGROUND: We sought to investigate risk factors for recurrence of vaginal intraepithelial neoplasia (VAIN) in the vaginal vault after laser vaporization. MATERIALS AND METHODS: Among 162 patients treated with laser vaporization in the vaginal vault after hysterectomy between March 1998 and August 2007, the clinical data of 68 patients were reviewed retrospectively. They underwent hysterectomy before laser vaporization due to the International Federation of Gynecology and Obstetrics (FIGO) stage Ia1-Ib1 cervical cancer (n = 17), cervical carcinoma in situ (CIS, n = 18), cervical intraepithelial neoplasia (CIN) (CIN I, n = 4; CIN II, n = 9; CIN III, n = 12), leiomyoma (n = 5) and endometrial polyp (n = 3). All patients were diagnosed as VAIN I (n = 24), VAIN II (n = 18) and VAIN III (n = 26) in the vaginal vault before laser vaporization. Risk factors for recurrence of VAIN in the vaginal vault after laser vaporization were investigated by multivariate logistic regression analysis. RESULTS: The median age was 48 years (range, 32-74 years), and the median duration of follow-up was 33 months (range, 10-115 months). Among all patients, 18 (26.5%) showed disease recurrence. Age (< or =48 years) and VAIN III in the vaginal vault before laser vaporization were risk factors for recurrence after laser vaporization (age < or =48 years, adjusted OR, 2.07, 95% CI, 1.18-7.01; VAIN III in the vaginal vault before laser vaporization, adjusted OR, 8.42, 95% CI, 1.63-18.56). CONCLUSION: Laser vaporization may be a feasible method for the treatment of VAIN in the vaginal vault after hysterectomy. However, age (< or =48 years) and VAIN III in the vaginal vault before laser vaporization should be considered as risk factors for recurrence after laser vaporization. PMID- 19291751 TI - Noninvasive laser vasectomy: preliminary ex vivo tissue studies. AB - BACKGROUND AND OBJECTIVES: Male sterilization (vasectomy) is more successful, safer, less expensive, and easier to perform than female sterilization (tubal ligation). However, female sterilization is more popular, primarily due to male fear of vasectomy complications (incision, bleeding, infection, and scrotal pain). The development of a completely noninvasive vasectomy technique may eliminate these concerns. MATERIALS AND METHODS: Ytterbium fiber laser radiation with a wavelength of 1,075 nm, average power of 11.7 W, 1-second pulse duration, 0.5 Hz pulse rate, and 3-mm-diameter spot was synchronized with cryogen cooling of the scrotal skin surface in canine tissue for a treatment time of 60 seconds. RESULTS: Vas thermal lesion dimensions measured 2.0+/-0.3 mm diameter by 3.0+/ 0.9 mm length, without evidence of skin damage. The coagulated vas bursting pressure measured 295+/-72 mm Hg, significantly higher than typical vas ejaculation pressures of 136+/- 29 mm Hg. CONCLUSIONS: Noninvasive thermal coagulation and occlusion of the vas was produced in an ex vivo canine tissue model. However, chronic in vivo animal studies will be necessary to optimize the laser/cooling treatment parameters and confirm long-term vas occlusion with absence of sperm in the ejaculate, before clinical application. PMID- 19291752 TI - In vivo effects of low level laser therapy on inducible nitric oxide synthase. AB - BACKGROUND AND OBJECTIVE: Low level laser therapy (LLLT) has been demonstrated to modulate inflammatory processes with evidence suggesting that treatment protocol, such as wavelength, total energy, and number of treatments determine the clinical efficacy. In this study, the effects of LLLT mediated by different wavelengths and continuous versus pulsed delivery mode were quantified in a transgenic murine model with the luciferase gene under control of the inducible nitric oxide synthase (iNOS) expression. STUDY DESIGN/MATERIALS AND METHODS: LLLT modulated iNOS gene expressed in the acute Zymosan-induced inflammation model is quantified using transgenic mice (FVB/N-Tg(iNOS-luc)). Here an energy density of 5 J cm(-2) at either 635, 660, 690, and 905 nm in continuous wave mode and at 905 nm for short pulse delivery were evaluated. Age of the animals was determined as additional modulating the inflammatory response and the LLLT efficacy for some treatment protocols. RESULTS: Animals younger than 15 weeks showed mostly reduction of iNOS expression, while older animals showed increased iNOS expression for some LLLT protocols. Intensity and time course of inducible nitric oxide expression was found to not only depend on wavelength, but also on the mode of delivery, continuous, or pulsed irradiation. CONCLUSION: LLLT exhibit different effects in induced inflammatory process according to different wavelengths and wave mode. Upregulation of iNOS gene following 905 nm pulsed wave suggests a different mechanism in activating the inflammatory pathway response when compared to the continuous wave. PMID- 19291753 TI - High contrast near-infrared polarized reflectance images of demineralization on tooth buccal and occlusal surfaces at lambda = 1310-nm. AB - BACKGROUND AND OBJECTIVES: Sound enamel manifests peak transparency in the near IR (NIR) at 1310-nm, therefore the near-IR is ideally suited for high contrast imaging of dental caries. The purpose of this study was to acquire images of early demineralized enamel on the buccal and occlusal surfaces of extracted human teeth using NIR reflectance imaging and compare the contrast of those images with the contrast of images taken using other methods. MATERIALS AND METHODS: Fifteen human molars were used in this in vitro study. Teeth were painted with a clear acid-resistant varnish, leaving two 2 mm x 2 mm windows on the buccal and occlusal surfaces of each tooth for demineralization. Artificial lesions were produced in the exposed windows after a 2-day exposure to a demineralizing solution at pH 4.5. Lesions were imaged using NIR transillumination, NIR and visible light reflectance, and fluorescence imaging methods. Crossed polarizers were used where appropriate to improve contrast. Polarization sensitive optical coherence tomography (PS-OCT) was also used to non-destructively assess the depth and severity of demineralization in each sample window. RESULTS: NIR reflectance imaging had the highest image contrast for both the buccal and occlusal groups and it was significantly higher contrast than visible light reflectance (P < 0.05). CONCLUSION: The results of the study suggest that NIR reflectance imaging is a promising new method for acquiring high contrast images of early demineralization on tooth surfaces. PMID- 19291754 TI - Performance assessment of novel side firing flexible optical fibers for dental applications. AB - BACKGROUND AND OBJECTIVE: Use of lasers in dentistry in root canals of teeth and in periodontal pockets for disinfection would be more effective if energy was delivered laterally. This study examined the energy emission profiles of fibers modified in various ways to enhance their lateral emissions for dental use. MATERIALS AND METHODS: Commercial optical fibers were altered by tube etching with hydrofluoric acid, modified tube etching (after removing the protective polyimide coating), alumina abrasive particle beams, and by etching and particle beams used in combination. Three free-running pulsed infrared lasers (Nd:YAG, Er:YAG, and Er,Cr:YSGG) were employed to test the modified fibers, with 25 fibers for each laser (modified or unmodified). Surface topography of fibers was examined using SEM. Laser emissions forward and laterally were measured, and thermally sensitive paper used to record the emission profiles. Visible tracing of emissions was undertaken using coaxial He-Ne or InGaAsP diode laser emissions. RESULTS: The etching/abrasion/etching combination gave a unique honeycomb surface configuration with grating-like properties, whilst etching alone gave a conical end. Conical and honeycomb tips showed greater lateral and lower forward emissions compared with plain fibers, with fourfold improvements in lateral emission. The most regular lateral emissions were from the honeycomb configuration. CONCLUSION: The honeycomb and the conical fiber modifications show dramatic improvements in lateral emissions. The unique emission profile obtained for the honeycomb fibers could play a significant role in increasing the efficiency of laser delivery for endodontic and periodontal applications in dentistry, as well as in other fields. PMID- 19291755 TI - Method for disruption and re-canalization of atherosclerotic plaques in coronary vessels with photothermal bubbles generated around gold nanoparticles. AB - BACKGROUND AND OBJECTIVES: Rapid and safe re-canalization of totally occluded coronary vessels, especially those with the calcified plaques, represent a challenge for cardiology. STUDY DESIGN/MATERIALS AND METHODS: We have suggested to employ photothermal microbubbles (PTMBs) that are generated around injected to plaque or thrombus gold nanoparticles with a short laser pulse for selective mechanical disruption and removal of the plaque tissue and without thermal and mechanical damage to arterial wall. PTMBs were generated in vitro around 30-250 nm gold spheres and with 10 nanoseconds laser pulse at 532 nm in three models: the layer of the living fibroblasts, the epoxy layers, and human arteries with plaques. RESULTS: In all three models, complete removal of the material was observed after 1-10 single laser pulses. The size of cleared zones (20-220 microm) was found to be 500-1,000 times bigger than the size of the nanoparticles applied. PTMB generation did not increase the temperature of the microenvironment outside PTMB and the debris size was below 2 microm. CONCLUSIONS: New proposed method for non-thermal mechanical and localized removal of plaque tissue with PTMB can provide safe and rapid re-canalization of totally occluded and calcified arteries without collateral damage. PMID- 19291756 TI - Low-energy laser irradiation promotes synovial fibroblast proliferation by modulating p15 subcellular localization. AB - BACKGROUND AND OBJECTIVE: Low-energy laser irradiation (low-level laser therapy) (LELI/LLLT/photobiomodulation) has been found to modulate various biological effects, especially those involved in promoting cell proliferation. Synovial fibroblasts are important in maintaining the homeostasis of articular joints and have strong chondrogenetic capacity. Here, we investigated the effect and molecular basis of LELI on synovial fibroblast proliferation. STUDY DESIGN/MATERIALS AND METHODS: HIG-82 rabbit synovial fibroblasts were cultured, and laser irradiation (660 nm) was applied at the power density of 40 mW/cm(2) for 2 minutes, corresponding to laser fluence of 4.8 J/cm(2). The effect of LELI on cell proliferation, cell cycle progression, and expression of cyclin-dependent kinase inhibitors (CKIs) were investigated. We also examined whether the effects of LELI on HIG-82 cell proliferation were affected by cAMP content, which is known to influence the cell cycle via inducing CKIs. RESULTS: LELI promoted HIG 82 synovial fibroblast proliferation and induced cytoplasmic localization of cyclin-dependent kinase inhibitor p15 (INK4B/CDKN2B). Moreover, the proliferation of HIG-82 synovial fibroblasts was reduced by cAMP, while cAMP inhibitor, SQ22536, induced p15 cytoplasmic localization and as a result, elevated synovial fibroblast proliferation was observed. In addition, the promotive effect of LELI induced HIG-82 synovial fibroblast proliferation was abolished by cAMP treatment. Our findings suggest that cAMP may be involved in the effect of LELI on synovial fibroblast proliferation. CONCLUSION: We revealed the effect and molecular link involved in synovial fibroblast proliferation induced by 660-nm LELI. Our study provides new insights into the mechanisms by which LELI has biological effects on synovial fibroblast proliferation. These insights may contribute to further investigation on biological effects and application of LELI in regenerative medicine. PMID- 19291757 TI - Three-dimensional study of pectoralis major muscle and tendon architecture. AB - A thorough understanding of the normal structural anatomy of the pectoralis major (PM) is of paramount importance in the planning of PM tendon transfers or repairs following traumatic PM tears. However, there is little consensus regarding the complex musculotendinous architecture of the PM in the anatomic or surgical literature. The purpose of this study is to model and quantify the three dimensional architecture of the pectoralis muscle and tendon. Eleven formalin embalmed cadaveric specimens were examined: five (2M/3F) were serially dissected, digitized, and modeled in 3D using Autodesk Maya; six (4M/2F) were dissected and photographed. The PM tendon consisted of longer anterior and shorter posterior layers that were continuous inferiorly. The muscle belly consisted of an architecturally uniform clavicular head (CH) and a segmented sternal head (SH) with 6-7 segments. The most inferior SH segment in all specimens was found to fold anteriorly forming a trough that cradled the inferior aspect of the adjacent superior segment. No twisting of either the PM muscle or tendon was noted. Within the CH, the fiber bundle lengths (FBL) were found to increase from superior to inferior, whereas the mean FBLs of SH were greatest in segments 3-5 found centrally. The mean lateral pennation angle was greater in the CH (29.4 +/- 6.9 degrees ) than in the SH (20.6 +/- 2.7 degrees ). The application of these findings could form the basis of future studies to optimize surgical planning and functional recovery of repair/reconstruction procedures. PMID- 19291758 TI - The ZMF: Is it a reliable intraoperative guide for the IOF? AB - Intraoperative localization of the inferior orbital fissure (IOF) is necessary when making an osteotomy across the zygomatic bone while performing an orbito zygomatic craniotomy. The zygomatico-facial foramen (ZMF) may serve as good reference point for locating the IOF. In this study, the position of the ZMF was assessed and its location in relation to the IOF was measured in 78 skulls. The ZMF was present in 83.3% of the skulls and when present, was related to the lateral end of the IOF in all cases. The mean distance of the ZMF from the IOF was 15.6 mm, and the mean distance between the ZMF and the fronto-zygomatic suture was 25.9 mm. In skulls where the ZMF was absent, the mean distance of the inferior orbital rim (at the level of IOF) from the fronto-zygomatic suture (20.7 mm) could be used for determining the position of the IOF. The IOF could thus be located approximately 15-16 mm medial to the ZMF in the same transverse plane. The location of the ZMF could also be used as a landmark for determining the inferior limit of the orbito-zygomatic craniotomy. PMID- 19291759 TI - Effects of sex, status, and mating cues on expected aggressive responses. AB - The effect of sex, status, and mating cues on expected aggression was examined via three scenario-based studies in which participants imagined themselves in a situation with a same-sex instigator of a provocation. Participants were randomly assigned to receive a scenario, which included one of two levels of status of instigator (high, low), one of two levels of attractiveness of the instigator (unattractive, attractive), and one of two levels of provocation (apology, insult). Sex and dispositional aggressivity were also included in a full factorial design. Based on evolutionary psychology ideas, we anticipated that status and attractiveness would differentially influence expected aggression for men vs. women. Participants in Experiment 1 were instructed to imagine that they were alone, whereas participants in Experiments 2 and 3 imagined themselves in a situation that included mating-related primes. In general expected aggression was greater for aggression-prone participants and under conditions of provocation and/or a high-status instigator. Experiments 2 and 3 found that, in the context of mate competition, sex differences in the effects of instigator provocation, status, and attractiveness emerged: greater aggressivity now only predicted more aggression for males but not females who were insulted; aggression was highest for females confronting an unattractive, high-status instigator and for males confronting an attractive, high-status instigator; females were more likely to aggress against a high-status instigator, regardless of being in a steady relationship or a first date situation, but males were only more likely to aggress against a high-status instigator in a first date situation. PMID- 19291760 TI - Kinesin-5 in Drosophila embryo mitosis: sliding filament or spindle matrix mechanism? AB - The Drosophila syncytial embryo uses multiple astral mitotic spindles that are specialized for rapid mitosis. The homotetrameric kinesin-5, KLP61F contributes to various aspects of mitosis in this system, all of which are consistent with it exerting outward forces on spindle poles. In principle, kinesin-5 could accomplish this by (i) sliding microtubules (MTs), minus end leading, relative to a static spindle matrix or (ii) crosslinking and sliding apart adjacent pairs of antiparallel interpolar (ip) MTs. Here, I critically review data on the biochemistry of purified KLP61F, its localization and dynamic properties within spindles, and quantitative modeling of KLP61F function. While a matrix-based mechanism may operate in some systems, the work tends to support the latter "sliding filament" mechanism for KLP61F action in Drosophila embryo spindles. Cell Motil. Cytoskeleton 2009. (c) 2009 Wiley-Liss, Inc. PMID- 19291761 TI - Comparing 6 and 10 sec exhalation time in exhaled nitric oxide measurements in children. AB - Standard exhalation time for Fractional exhaled Nitric Oxide (FeNO) measurements is 10 sec but this is difficult for young children. Recommended exhalation time for children is 6 sec, but this was never substantiated in literature. We aimed to investigate the agreement between FeNO values measured with exhalation times of 6 and 10 sec and the preference of children for either method. The study population comprised children aged 5-17 years visiting the Pediatric Pulmonology outpatient clinic. FeNO values, measured during 6 (FeNO-6) and 10 (FeNO-10) sec (random order) using the single-breath online (SBOL) technique, were compared. Preferences for exhalation times were related to FVC values. Ninety-eight children (mean age 10.6 years) were included. Median FeNO-6 (15.2 ppb) and FeNO 10 (13.6 ppb) did not differ significantly (P = 0.259). Mean difference between FeNO-6 and FeNO-10 was -0.3 ppb, limits of agreement ranging from -5.8 ppb to +5.3 ppb. Sixty percent of children with a Forced Vital Capacity (FVC) less than 3 L preferred the FeNO-6 method. We found good agreement between FeNO-6 and FeNO 10, so they can be used interchangeably. An exhalation time of 6 sec was preferred by the majority of subjects with a FVC below 3 L. PMID- 19291762 TI - GAPO syndrome with the novel features of pulmonary hypertension, ankyloglossia, and prognathism. PMID- 19291763 TI - Interventions to improve patient education regarding multifactorial genetic conditions: a systematic review. AB - The careful education of patients with complex genetic disease is essential. However, healthcare providers often have limited time to spend providing thorough genetic education. Furthermore, the number of healthcare professionals possessing strong genetics training may be inadequate to meet increasing patient demands. Due to such constraints, several interventions have been investigated over the past decade to identify potential resources for the facilitation of this specific type of patient education. This systematic literature review of these interventions for patient education attempts to elucidate the answer to the question: is there sufficient evidence for best practice for delivering genetic information to patients with multifactorial conditions? The various interventions (CD-ROM, group counseling, video/decision aid, and miscellaneous) were analyzed in terms of quality criteria and achievement of specific outcomes and were rated according to the Stetler model for evidence-based practice. Seven main outcomes were evaluated: (1) objective and subjective knowledge assessment, (2) psychological measures (general anxiety, depression, stress, cancer worry), (3) satisfaction/effectiveness of intervention, (4) time spent in counseling (time spent on basic genetic information vs. specific concerns), (5) decision making/intent to undergo genetic testing, (6) treatment choice and value of that choice, and, finally (7) risk perception. Overall, the computer interventions resulted in more significant findings that were beneficial than any other category followed by the video category, although the group and miscellaneous categories did not measure all of the outcomes reported by the other two categories. Nevertheless, while these groups had neutral or negative findings in some of the outcomes, the computer intervention group showed significant improvement in genetics knowledge, psychological measures, satisfaction/effectiveness, time spent with counselor, and decision/intent to undergo testing. PMID- 19291764 TI - RNF135 mutations are not present in patients with Sotos syndrome-like features. PMID- 19291765 TI - Performance of plane-wave-based LDA+U and GGA+U approaches to describe magnetic coupling in molecular systems. AB - This work explores the performance of periodic plane wave density functional theory calculations with an on-site Coulomb correction to the standard LDA and GGA exchange-correlation potential--commonly used to describe strongly correlated solids--in describing the magnetic coupling constant of a series of molecular compounds representative of dinuclear Cu complexes and of organic diradicals. The resulting LDA+U or GGA+U formalisms, lead to results comparable to experiment and to those obtained by means of standard hybrid functionals provided that the value of the U parameter is adequately chosen. Hence, these methods offer an alternative efficient computational scheme to correct LDA and GGA approaches to adequately describe the electronic structure and magnetic coupling in large molecular magnetic systems, although at the expenses of introducing an empirical (U) parameter. For all investigated copper dinuclear systems, the LDA+U and GGA+U approaches lead to an improvement in the description of magnetic properties over the original LDA and GGA schemes with an accuracy similar to that arising from the hybrid B3LYP functional, by increasing the on-site Coulomb repulsion with a moderate U value. Nevertheless, the introduction of an arbitrary U value in the 0 10 eV range most often provides the correct ground-state spin distribution and the correct sign of the magnetic coupling constant. PMID- 19291766 TI - Fragile X screening: attitudes of genetic health professionals. AB - Although genetic health professionals (GHP) are major stakeholders in developing and implementing fragile X (FrX) testing and screening guidelines, their attitudes about FrX testing and population screening are virtually absent in the literature. A survey was conducted of physician geneticists (geneticists) and genetic counselors (GC). The survey addressed GHP's attitudes towards (1) prenatal FrX carrier screening; (2) pre- and full mutation screening of male and female newborns; (3) the single best time for FrX screening over the lifespan; and (4) their willingness to test a normally developing child with a positive family history. Surveys were completed by 30% (273/894) of eligible GHP. Attitudes of geneticists and GC were mostly indistinguishable. The single most favored screening approaches were (1) preconception screening targeted at women with a positive family history (43%); and (2) universal preconception screening (29%). While only 6% and 11% declared universal prenatal and universal newborn screening (NBS) as the ideal time respectively, 73% and 60% respectively would support such programs. GHP would design a NBS program to test male and female infants and to identify both pre- and full mutations. Over half would agree to order FrX testing on some normally developing children with a positive family history. In expanding FrX testing and screening to low risk individuals, GHP prefer preconception screening as the single best time. The majority also support prenatal screening and NBS. If NBS were to be introduced, GHP prefer screening to identify boys and girls with both pre- and full mutations. PMID- 19291767 TI - A novel genetic syndrome characterized by pediatric cataract, dysmorphism, ectodermal features, and developmental delay in an indigenous Australian family. AB - A novel syndrome initially presenting with cataract and developmental delay within an Indigenous Australian family is described. We present the extended four generation pedigree and describe in detail the phenotypic appearance of five clearly affected male second cousins in this family. The common features of these children include developmental delay, short stature, cortical cataract, facial dysmorphism, clinodactyly, thin hair and an erythematous skin rash. Initial inspection of the pedigree suggested an inherited disorder with possible X-linked inheritance. However, a thorough scan of the X chromosome failed to reveal linkage. This family represents a new syndrome of familial cataract, dysmorphic features, short stature and developmental delay with probable autosomal inheritance and variable expressivity. PMID- 19291768 TI - Autosomal dominant inheritance in a large family with focal facial dermal dysplasia (Brauer-Setleis syndrome). AB - Focal facial dermal dysplasia (FFDD) (OMIM 227260) is a rare ectodermal disorder characterized by congenital bitemporal scar-like depressions resembling forceps marks and variable additional facial manifestations. No gene defects or gene loci for FFDD are known to date. We report on a large multi-generational German family with typical characteristics of FFDD and provide a detailed clinical description of four affected individuals. They had large bitemporal discolored dermal depressions, sparse lateral eyebrows, abnormal eyelashes, and dysplastic and low set ears. Three of the four affected individuals had congenital horizontal nystagmus, which had hitherto only been reported in a single patient with FFDD. In contrast to previous assumptions about an autosomal recessive etiology of this disorder, this family provides further evidence that FFDD is inherited in an autosomal dominant mode. Although this family is not large enough to yield significant results in linkage analysis, it may, in combination with other families, contribute to the identification of a gene locus for this intriguing ectodermal disorder. PMID- 19291769 TI - Molecular cytogenetic characterization of an interstitial de novo 13q deletion in a 3-month-old with severe pediatric gastroesophageal reflux. AB - Gastroesophageal reflux (GER) occurs when gastric contents travel back into the esophagus through the esophageal sphincter. GER is very common in infants with most growing out of it, but some continue to have chronic symptoms throughout childhood and adulthood. A gene for severe pediatric gastroesophageal reflux disease (GERD) was identified by linkage analysis and was mapped to chromosome 13. We report here a de novo interstitial deletion of chromosome 13 in a 3-month old biracial male who presented to the emergency room with severe GER and failure to thrive. Chromosome analysis showed an interstitial deletion of chromosome 13, with the karyotype reported as 46, XY, del(13)(q12.3q14.1). BAC-FISH analysis demonstrated that the deletion encompasses 12.3 Mb and does involve the GERD1 locus. The GERD1 locus has been mapped to a 9-cM interval between the markers CAGR1 and D13S263, both of which are deleted in our patient. We propose that the GER phenotype in our patient is due to a haploinsufficiency of GERD1. PMID- 19291770 TI - A patient with Baller-Gerold syndrome and midline NK/T lymphoma. AB - Three autosomal recessive disorders are associated with mutations in the RECQL4 gene: Rothmund-Thomson syndrome (RTS), Baller-Gerold syndrome (BGS), and RAPADILINO syndrome. BGS is characterized by two major clinical abnormalities: craniosynostosis and preaxial limb anomalies but not cancer development. We performed RECQL4 mutation detection in a patient with BGS and several clinical signs of RTS who developed a midline NK/T-cell lymphoma. Sequencing was used to identify RECQL4 mutations, and RNA analysis was used to examine expression of mRNA in leukocytes. The patient was found to be compound heterozygous for two mutations in exon 15, namely c.[2492_2493delAT] + c.[2506_2518del13bp]. We found that only the allele with 13 bp deletion was expressed in blood leukocytes. Our patient showed severe phenotypic abnormalities, with clinical signs of both BGS and RTS. She developed an extranodal NK/T-cell lymphoma, which is extremely rare in children of her age and is the first described case of BGS with development of a cancer. This case of a RECQL4-related disorder highlights the significant phenotypic overlap between the classically delineated RECQL4-associated syndromes and questions the need to redefine or combine these clinical entities. PMID- 19291771 TI - Somatic mosaicism for the SALL1 mutation p.Ser371X in full-blown Townes-Brocks syndrome with Duane anomaly. PMID- 19291772 TI - A ZRS duplication causes syndactyly type IV with tibial hypoplasia. PMID- 19291773 TI - Xq13.2q21.1 duplication encompassing the ATRX gene in a man with mental retardation, minor facial and genital anomalies, short stature and broad thorax. AB - In a man with severe mental retardation, minor facial and genital anomalies, disproportionate short stature and a broad thorax, we identified a de novo Xq13.2q21.1 duplication by array CGH. This 7 Mb duplication encompasses 23 known genes, including the X-linked mental retardation (XLMR) genes ATRX and SLC16A2. The phenotype of this patient is similar to that described in more than 10 previously reported patients with overlapping Xq duplications. Detailed comparison of the clinical characteristics and the function of the genes located in the commonly duplicated regions of these patients led us to the hypothesis that an increased dosage of ATRX and perhaps of other genes is involved in the pathogenetic mechanism of this XLMR phenotype, including mental retardation, short stature, and genital abnormalities comprising cryptorchidism and/or a small penis. PMID- 19291774 TI - Indirect action of tumor necrosis factor-alpha in liver injury during the CD8+ T cell response to an adeno-associated virus vector in mice. AB - CD8+ T cells can cause hepatocellular injury by two distinct mechanisms. In addition to their direct cytotoxic effect, there is also collateral liver injury, which occurs when cells are killed in an antigen-independent manner. Whereas immune effector cytokines interferon-gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) have both been implicated in various forms of hepatitis, their respective roles in direct and/or collateral liver damage remains unclear. In order to investigate these elements of liver injury, we developed a new experimental model of CD8+ T-cell-mediated hepatitis based on an adeno-associated virus-based gene therapy vector. This vector is used to deliver antigen to hepatocytes, and CD8+ T cells specific for the vector-encoded transgene are adoptively transferred to produce liver immunopathology. In this experimental model, CD8+ T-cell IFNgamma acts on Kupffer cells, inducing TNFalpha secretion and liver injury. Both IFNgamma and TNFalpha are important in this injury process, but TNFalpha acts as an autocrine amplifier of Kupffer cell function, rather than as a direct effector of hepatocellular damage. CONCLUSIONS: TNFalpha indirectly promotes liver damage and is not a direct hepatotoxic agent. IFNgamma also indirectly contributes to liver injury through Kupffer cell activation while, in parallel, directly promoting hepatitis through induction of hepatocyte major histocompatability complex class I. In principle, it may be possible to ameliorate this immunopathologic indirect mechanism by developing therapies that target Kupffer cells, without impairing CD8+ T-cell-mediated antiviral immunity. This would have great therapeutic potential in chronic viral hepatitis. PMID- 19291775 TI - The missense mutation G12D in connexin30.3 can cause both erythrokeratodermia variabilis of Mendes da Costa and progressive symmetric erythrokeratodermia of Gottron. AB - Progressive symmetric erythrokeratoderma of Gottron (PSEK) is commonly distinguished from erythrokeratodermia variabilis Mendes da Costa (EKV). However, conclusive proof that the disorders are identical is still lacking. We performed mutation analysis and microsatellite haplotyping in two independently referred patients with PSEK and three patients from a previously published family with EKV. All patients had the same mutation in the GJB4 gene causing the amino acid substitution p.Gly12Asp (G12D). Haplotype analysis showed that all five patients had the same allelic haplotype over 2 Mb covering the disease locus. Apparently, the same GJB4 mutation may cause either an EKV or a PSEK phenotype. A single ancestral founder might have introduced EKV in the Netherlands. PMID- 19291776 TI - Familial upper eyelid coloboma with ipsilateral anterior hairline abnormality: two new reports of MOTA syndrome. AB - We describe two patients with upper eyelid coloboma, hypertelorism, disruption of the eyebrow, and anterior hairline anomaly. The phenotype in our patients is consistent with Manitoba Oculotrichoanal syndrome (MOTA syndrome, OMIM 248450), which comprises a variable spectrum of eyelid malformations ranging from cryptophthalmos to upper eyelid colobomas; aberrant ipsilateral anterior hairline, and anal anomalies. It was first described in members of the indigenous population of the Island Lake region of Northern Manitoba, Canada. Autosomal recessive inheritance is demonstrated in these families and single-gene etiology has been proposed. This constellation of anomalies also corresponds to those arising from the Number 10 cleft in Tessier's anatomical classification of clefting malformations. Tessier Number 10 clefts are the rarest of the 15 craniofacial clefting malformations first described by Tessier [Tessier (1976); J Maxillofac Surg 4:69-92]. They have only ever been reported as sporadic occurrences and the underlying etiology is thought to be environmental. We believe the phenotype in our patients and in those previously described with MOTA syndrome represents a disorder of craniofacial clefting; specifically, one that occurs along the tissue planes of the Tessier Number 10 cleft. The familial clustering of these facial features and their variable association with other congenital anomalies supports a genetic rather than environmental cause. The mapping of the gene for this syndrome is likely to involve a combined functional and positional approach with a focus on candidate genes involved in craniofacial development. PMID- 19291777 TI - The phenotype of persons having mosaicism for trisomy 21/Down syndrome reflects the percentage of trisomic cells present in different tissues. AB - Little is known about the pathogenesis of the phenotype in individuals with trisomy 21 mosaicism and Down syndrome. The primary goal of this study was to identify factors contributing to the observed phenotypic variation by evaluating 107 individuals having trisomy 21 mosaicism. To investigate a potential "threshold" effect due to trisomic imbalance, lymphocyte and buccal mucosa nuclei were scored using FISH. Overall, buccal cells showed a significantly higher frequency of trisomy than lymphocytes (P < 0.0001). Using latent class analysis, two phenotypic classes were identified based on the clinical findings of the propositi. Patients from class 1 had significantly fewer traits and a lower percentage of trisomic cells (mean of 37.3% lymphocytes; 34.5% buccal mucosa cells) when compared to those stratified into class 2 (54.0% lymphocytes; 53.4% buccal mucosa cells). Tissue-specific influences were also detected, with buccal mucosa trisomy levels being significantly correlated with IQ (P = 0.0094; both ectodermal derivatives), while congenital heart defects were significantly correlated with lymphocytes (P = 0.0286; both mesodermal embryonic derivatives). In conclusion, allowing for the distinction of two groups, we observed variation in phenotype, associated with the percentage of trisomic cells. We also observed tissue-specific effects on phenotype. The results of this study should enable geneticists and other health care professionals to provide information regarding optimal diagnostic approaches and anticipated clinical outcomes. PMID- 19291778 TI - Epidemiology of acephalus/acardius monozygotic twins: new insights into an epigenetic causal hypothesis. AB - Apart from a series of 10 acephalus/acardius (Ac/Ac) cases described from a pathological point of view, and the analysis of a review of published cases, we have been unable to find any epidemiological studies on Ac/Ac. Using data from the Spanish Collaborative Study of Congenital Malformations (ECEMC), we present here what seems to be the first epidemiological analysis of a consecutive series of the Ac/Ac type of monozygotic twins (MZT). Among a total of 2,281,604 consecutive births, 11 cases of Ac/Ac MZT were detected, giving a frequency of 0.48 per 100,000 births. However, we consider the period 1980-1985 as the baseline for our data, as in this period voluntary termination of pregnancy was not possible in Spain, and the frequency of Ac/Ac MZT was 0.49 per 100,000 births. Nonetheless, this frequency should be considered as a minimal estimation. The characteristics of these Ac/Ac cases indicate that they are more frequent in males (sex ratio 2.67). In addition, gestational age in Ac/Ac cases was 2.41 and 3.12 weeks lower than in malformed and control twins, respectively. Similarly, their mothers are 4.54 and 4.68 years younger than mothers of separate malformed and control twins, respectively. To understand the biological basis behind the occurrence of MZT in the context of recent observations, we evaluate the hypothesis that the epigenetic processes involved in the early cleavage of the embryo, and in blastocyst formation during development, may be implicated in twinning. PMID- 19291779 TI - Assessment and validation of the new capsule endoscopy Crohn's disease activity index (CECDAI): what difference does it make? PMID- 19291780 TI - Fecal calprotectin and lactoferrin for the prediction of inflammatory bowel disease relapse. AB - BACKGROUND: The purpose of the study was to determine the role of fecal calprotectin and lactoferrin in the prediction of inflammatory bowel disease relapses, both in patients with ulcerative colitis (UC) and Crohn's disease (CD), in a large, long-term, follow-up study. METHODS: The prospective multicenter study included CD and UC patients who had been in clinical remission for 6 months. At baseline, patients provided a single stool sample for calprotectin and lactoferrin determination. Follow-up was 12 months in patients showing no relapse and until activity flare in relapsing patients. RESULTS: In all, 163 patients (89 CD, 74 UC) were included. Twenty-six patients (16%) relapsed during follow-up. Calprotectin concentrations in patients who suffered a relapse were higher than in nonrelapsing patients (239 +/- 150 versus 136 +/- 158 microg/g; P < 0.001). Relapse risk was higher in patients having high (>150 microg/g) calprotectin concentrations (30% versus 7.8%; P < 0.001) or positive lactoferrin (25% versus 10%; P < 0.05). Fecal calprotectin (>150 microg/g) sensitivity and specificity to predict relapse were 69% and 69%, respectively. Corresponding values for lactoferrin were 62% and 65%, respectively. The area under the receiver operating characteristic curve to predict relapse using calprotectin determination was 0.73 (0.69 for UC and 0.77 for CD). Better results were obtained when only colonic CD disease or only relapses during the first 3 months were considered (100% sensitivity). High fecal calprotectin levels or lactoferrin positivity was associated with clinical relapse in Kaplan-Meier survival analysis, and both fecal tests were associated with relapse in the multivariate analysis. CONCLUSIONS: Fecal calprotectin and lactoferrin determination may be useful in predicting impending clinical relapse-especially during the following 3 months-in both CD and UC patients. PMID- 19291781 TI - Effects of infliximab therapy on abdominal fat and metabolic profile in patients with Crohn's disease. AB - BACKGROUND: Tumor necrosis factor is an adipocytokine possessing a well established lipolytic effect. In Crohn's disease (CD) patients, infliximab therapy may thus result in visceral fat accumulation, which is associated with an increased risk of metabolic syndrome. METHODS: A total of 132 CD patients were investigated. In a first prospective study, magnetic resonance imaging (MRI) quantification of subcutaneous and visceral abdominal fat was performed before and 8 weeks after initiation of infliximab induction therapy (5 mg/kg at weeks 0, 2, and 6) in 21 responding patients treated for perianal disease. In a second prospective study, fasting glycemia, glycated hemoglobin (HbA1c), HDL, LDL, and total cholesterol and triglyceride levels were assessed in 111 responding patients receiving infliximab infusions every 8 weeks, with a mean follow-up of 41 weeks. RESULTS: A significant homogeneous 18% increase in total abdominal fat was observed in the 21 CD patients after infliximab induction therapy (P = 0.027), independently of body mass index evolution. Infliximab maintenance therapy was associated with a decrease in glycemia (P < 0.0001) and HbA1c (P = 0.0005) concentrations, together with an increase in both total cholesterol (P = 0.02) and HDL cholesterol (P = 0.008) concentrations. All glycemic and lipid parameters remained within the normal range throughout the study. CONCLUSIONS: Infliximab induction therapy is associated with a significant increase in abdominal fat tissue in CD patients. Infliximab maintenance therapy has no deleterious effects on lipid profile and is accompanied by a decrease in glycemia and HbA1c concentrations, probably by reversing the impairment of tumor necrosis factor-induced insulin-mediated glucose uptake. PMID- 19291782 TI - Red cell exchange transfusion for babesiosis in Rhode Island. AB - We report four cases of clinically severe tick borne babesiosis treated with chemotherapy and adjunctive red cell exchange (RCE) at two Rhode Island hospitals from 2004 to 2007. All RCE procedures were performed using a Cobe Spectra device and were well tolerated without complications. The volume of allogeneic red cells used in the exchange was determined using the algorithm in the apheresis device with the input variables of preprocedure hematocrit, weight, height, an assumed allogeneic red cell hematocrit of 55 and a desired post procedure hematocrit of 27. The preprocedure level of parasitemia varied between 2.4% and 24% and the postprocedure level of parasitemia between 0.4 and 5.5% with an average overall percent reduction in parasitemia of 74%. Retrospectively, application of a new formula to calculate red cell mass appeared to correlate better with the percent reduction in parasitemia. Previous reports of RCE in babesiosis were reviewed. The reported reduction in parasitemia varied from 50% to >90%. Although a preprocedure level of parasitemia of 10% is sometimes used as a threshold for RCE in clinically severe babesiosis, this threshold does not have a firm empirical basis. No postprocedure desired level of parasitemia is indicated nor the mass of allogeneic red cells needed to achieve such a level. We conclude that current estimates of the dose of allogeneic red cells used in RCE are probably inaccurate, advocate a new formula to estimate this dose and suggest that a 90% reduction in parasitemia should be the minimally desired target of RCE in babesiosis. PMID- 19291783 TI - Lidocaine prevents referred hyperalgesia associated with cystitis. AB - AIMS: Lidocaine produces analgesia by inhibiting excitation of nerve endings or blocking impulse conduction in peripheral nerves. This study was performed to determine whether intrathecal or intravesical administration of lidocaine prior, or subsequent, to induction of chemical cystitis in rats would block referred mechanical hyperalgesia. METHODS: Intrathecal or intravesical lidocaine was administered 15 (intrathecal) or 30 (intravesical) min before intravesical instillation of saline or 1 mM acrolein (400 microl) or 4 hr after saline or acrolein instillation in female Wistar rats. Mechanical sensitivity of hind paws was determined at 24 hr prior to any treatment (baseline) and, 4, 24, and 48 hr after intravesical instillation of acrolein or saline. Also, nerve growth factor (NGF) content was measured in bladder and dorsal root ganglia (DRG). RESULTS: Pre treatment with intrathecal or intravesical lidocaine attenuated acrolein-induced referred mechanical hyperalgesia of the hind paws. Lidocaine administered after acrolein instillation did not alter referred hyperalgesia. Lidocaine treatment prior to or after induction of cystitis reduced NGF content in the bladder. CONCLUSIONS: These results indicate that pre-treatment with lidocaine attenuates referred hyperalgesia associated with cystitis. Lidocaine treatment 4 hr after induction of cystitis failed to prevent referred hyperalgesia despite a similar decrease in bladder NGF. Neurourol. Urodynam. (c) 2009 Wiley-Liss, Inc. PMID- 19291784 TI - SAFE biopsy: a validated method for large-scale staging of liver fibrosis in chronic hepatitis C. AB - The staging of liver fibrosis is pivotal for defining the prognosis and indications for therapy in hepatitis C. Although liver biopsy remains the gold standard, several noninvasive methods are under evaluation for clinical use. The aim of this study was to validate the recently described sequential algorithm for fibrosis evaluation (SAFE) biopsy, which detects significant fibrosis (> or =F2 by METAVIR) and cirrhosis (F4) by combining the AST-to-platelet ratio index and Fibrotest-Fibrosure, thereby limiting liver biopsy to cases not adequately classifiable by noninvasive markers. Hepatitis C virus (HCV) patients (2035) were enrolled in nine locations in Europe and the United States. The diagnostic accuracy of SAFE biopsy versus histology, which is the gold standard, was investigated. The reduction in the need for liver biopsies achieved with SAFE biopsy was also assessed. SAFE biopsy identified significant fibrosis with 90.1% accuracy (area under the receiver operating characteristic curve = 0.89; 95% confidence interval, 0.87-0.90) and reduced by 46.5% the number of liver biopsies needed. SAFE biopsy had 92.5% accuracy (area under the receiver operating characteristic curve = 0.92; 95% confidence interval, 0.89-0.94) for the detection of cirrhosis, obviating 81.5% of liver biopsies. A third algorithm identified significant fibrosis and cirrhosis simultaneously with high accuracy and a 36% reduction in the need for liver biopsy. The patient's age and body mass index influenced the performance of SAFE biopsy, which was improved with adjusted Fibrotest-Fibrosure cutoffs. Two hundred two cases (9.9%) had discordant results for significant fibrosis with SAFE biopsy versus histology, whereas 153 cases (7.5%) were discordant for cirrhosis detection; 71 of the former cases and 56 of the latter cases had a Fibroscan measurement within 2 months of histological evaluation. Fibroscan confirmed SAFE biopsy findings in 83.1% and 75%, respectively. CONCLUSION: SAFE biopsy is a rational and validated method for staging liver fibrosis in hepatitis C with a marked reduction in the need for liver biopsy. It is an attractive tool for large-scale screening of HCV carriers. PMID- 19291785 TI - Increased intestinal permeability and tight junction alterations in nonalcoholic fatty liver disease. AB - The role played by the gut in nonalcoholic fatty liver disease (NAFLD) is still a matter of debate, although animal and human studies suggest that gut-derived endotoxin may be important. We investigated intestinal permeability in patients with NAFLD and evaluated the correlations between this phenomenon and the stage of the disease, the integrity of tight junctions within the small intestine, and prevalence of small intestinal bacterial overgrowth (SIBO). We examined 35 consecutive patients with biopsy-proven NAFLD, 27 with untreated celiac disease (as a model of intestinal hyperpermeability) and 24 healthy volunteers. We assessed the presence of SIBO by glucose breath testing (GBT), intestinal permeability by means of urinary excretion of (51)Cr-ethylene diamine tetraacetate ((51)Cr-EDTA) test, and the integrity of tight junctions within the gut by immunohistochemical analysis of zona occludens-1 (ZO-1) expression in duodenal biopsy specimens. Patients with NAFLD had significantly increased gut permeability (compared with healthy subjects; P < 0.001) and a higher prevalence of SIBO, although both were lower than in the untreated celiac patients. In patients with NAFLD, both gut permeability and the prevalence of SIBO correlated with the severity of steatosis but not with presence of NASH. CONCLUSIONS: Our results provide the first evidence that NAFLD in humans is associated with increased gut permeability and that this abnormality is related to the increased prevalence of SIBO in these patients. The increased permeability appears to be caused by disruption of intercellular tight junctions in the intestine, and it may play an important role in the pathogenesis of hepatic fat deposition. PMID- 19291786 TI - Golgi phosphoprotein 2 (GOLPH2) expression in liver tumors and its value as a serum marker in hepatocellular carcinomas. AB - Hepatocellular carcinomas (HCCs) and bile duct carcinomas (BDCs) have a poor prognosis. Therefore, surveillance strategies including sensitive and specific serum markers for early detection are needed. Recently, Golgi Phosphoprotein 2 (GOLPH2) has been proposed as a serum marker for HCC, but GOLPH2 expression data in liver tissues was not available. Using tissue microarrays and immunohistochemistry, we semiquantitatively analyzed GOLPH2 protein expression in patients with HCC (n = 170), benign liver tumors (n = 22), BDC (n = 114) and normal liver tissue (n = 105). A newly designed sandwich enzyme-linked immunoassay (ELISA) was used to analyze GOLPH2 levels in the sera of patients with HCC (n = 62), hepatitis C virus (HCV) (n = 29), BDC (n = 10), and healthy control persons (n = 12). By immunohistochemistry 121/170 (71%) of HCC showed strong GOLPH2 expression, which was significantly associated with a higher tumor grade (P = 0.01). A total of 97/114 (85%) BDCs showed a strong GOLPH2 expression which proved to be an independent prognostic factor for overall survival (P < 0.05). Serum levels of GOLPH2 measured by ELISA were significantly elevated in patients with HCC with underlying HCV infection (median 18 mg/L, P < 0.05) and patients with BDC (median = 14.5 mg/L, P < 0.01) in comparison to healthy controls (median 4 mg/L). CONCLUSION: GOLPH2 protein is highly expressed in tissues of HCC and BDC. GOLPH2 protein levels are detectable and quantifiable in sera by ELISA. In patients with hepatitis C, serial ELISA measurements in the course of the disease appear to be a promising complementary serum marker in the surveillance of HCC. GOLPH2 should be further evaluated as a serum tumor marker in BDC on a larger scale. PMID- 19291787 TI - Evolution of hepatic steatosis in patients with advanced hepatitis C: results from the hepatitis C antiviral long-term treatment against cirrhosis (HALT-C) trial. AB - Hepatic steatosis is a common histologic feature in patients with chronic hepatitis C (CHC) but there are no large longitudinal studies describing the progression of steatosis in CHC. We examined changes in steatosis on serial biopsies among CHC patients participating in the Hepatitis C Antiviral Long-term Treatment against Cirrhosis (HALT-C) Trial. All 1050 patients in the trial had advanced fibrosis at baseline biopsy and were documented not to have had a sustained virological response to peginterferon and ribavirin. Most (94%) patients had genotype 1 infection. At least one protocol follow-up biopsy was read on 892 patients, and 699 had the last biopsy performed 3.5 years after randomization. At enrollment, 39% had cirrhosis and 61% had bridging fibrosis; 18%, 41%, 31%, and 10% had steatosis scores of 0, 1, 2, and 3 or 4, respectively. The mean steatosis score decreased in the follow-up biopsies in both the interferon-treated patients and controls with no effect of treatment assignment (P = 0.66). A decrease in steatosis score by > or =1 point was observed in 30% of patients and was associated with both progression to cirrhosis and continued presence of cirrhosis (P = 0.02). Compared to patients without a decrease in steatosis, those with a decrease in steatosis had worse metabolic parameters at enrollment, and were more likely to have a decrease in alcohol intake, improvement in metabolic parameters, and worsening liver disease (cirrhosis, esophageal varices, and deterioration in liver function). CONCLUSION: Serial biopsies demonstrated that in patients with CHC, steatosis recedes during progression from advanced fibrosis to cirrhosis. Decreased alcohol intake and improved metabolic parameters are associated with a decline in steatosis and may modulate hepatitis C progression. PMID- 19291788 TI - Fibrin accumulation plays a critical role in the sensitization to lipopolysaccharide-induced liver injury caused by ethanol in mice. AB - The early stages of alcohol-induced liver injury involve chronic inflammation. Whereas mechanisms by which this effect is mediated are not completely understood, it is hypothesized that enhanced sensitivity to circulating lipopolysaccharide (LPS) contributes to this process. It has recently been shown that ethanol induces activation of plasminogen activator inhibitor-1 (PAI-1). PAI 1 causes fibrin accumulation in liver by inhibiting degradation of fibrin (fibrinolysis). LPS also enhances fibrin accumulation by activating the coagulation cascade. It was therefore hypothesized that ethanol will synergistically increase fibrin accumulation caused by LPS, enhancing liver damage. Accordingly, the effect of ethanol pretreatment on LPS-induced liver injury and fibrin deposition was determined in mice. Ethanol enhanced liver damage caused by LPS, as determined by plasma parameters and histological indices of inflammation and damage. This effect was concomitant with a significant increase in PAI-1 expression. Extracellular fibrin accumulation caused by LPS was also robustly increased by ethanol preexposure. Coadministration of the thrombin inhibitor hirudin or the MEK (mitogen-activated protein kinase) inhibitor U0126 significantly attenuated the enhanced liver damage caused by ethanol preexposure; this protection correlated with a significant blunting of the induction of PAI-1 caused by ethanol/LPS. Furthermore, thrombin/MEK inhibition prevented the synergistic effect of ethanol on the extracellular accumulation of fibrin caused by LPS. Similar protective effects on fibrin accumulation were observed in tumor necrosis factor receptor 1 (TNFR-1)(-/-) mice or in wild-type injected with PAI-1 inactivating antibody. CONCLUSION: These results suggest that enhanced LPS induced liver injury caused by ethanol is mediated, at least in part, by fibrin accumulation in livers, mediated by an inhibition of fibrinolysis by PAI-1. These results also support the hypothesis that fibrin accumulation may play a critical role in the development of early alcohol-induced liver injury. PMID- 19291789 TI - Fatty liver is associated with insulin resistance, risk of coronary heart disease, and early atherosclerosis in a large European population. AB - Patients with fatty liver (FL) disease have a high risk of developing diabetes and cardiovascular diseases. The aim was to evaluate the association between FL, insulin resistance (IR), coronary heart disease (CHD) risk, and early atherosclerosis in a large European population (RISC Study). In 1,307 nondiabetic subjects (age 30-60 years) recruited at 19 centers, we evaluated liver enzymes, lipids, insulin sensitivity (by euglycemic-hyperinsulinemic clamp), glucose tolerance (by 75 g oral glucose tolerance test), carotid atherosclerosis as intima media thickness (IMT), CHD risk by the Framingham Heart study prediction score, and physical activity (by accelerometer). The presence of FL was estimated using the fatty liver index (FLI; >60, likelihood >78% presence FL; FLI <20 likelihood >91% absence of FL). Subjects were divided into three groups: G1: FLI <20 (n = 608); G3: FLI >60 (n = 234), G2: intermediate group (n = 465). Compared to G1, G3 included more men (70% versus 24%) and people with impaired glucose tolerance (23% versus 5%). IMT increased with FLI (G3 = 0.64 +/- 0.08 versus G1 = 0.58 +/- 0.08 mm, P < 0.0001). FLI was associated with increased CHD risk (r = 0.48), low-density lipoprotein cholesterol (r = 0.33), alanine aminotransferase (r = 0.48), aspartate aminotransferase (r = 0.25), systolic blood pressure (r = 0.39) and IMT (r = 0.30), and reduced insulin sensitivity (r = -0.43), high density lipoprotein cholesterol (r = -0.50), adiponectin (r = -0.42), and physical activity (r = -0.16, all P < 0.0001). The correlations hold also in multivariate analysis after adjusting for age, gender, and recruiting center. CONCLUSION: In middle-age nondiabetic subjects, increased IMT, CHD risk, and reduced insulin sensitivity are associated with high values of FLI. PMID- 19291790 TI - Retreating chronic hepatitis C with daily interferon alfacon-1/ribavirin after nonresponse to pegylated interferon/ribavirin: DIRECT results. AB - Up to 50% of patients with chronic hepatitis C fail to respond to initial therapy with pegylated interferon (PEG-IFN) and ribavirin (RBV). With unsuccessful viral eradication, these patients remain at risk for developing progression of their liver disease. Retreatment with PEG-IFN/RBV yields sustained virologic response (SVR) rates that are under 10%. A wholly synthetic interferon, interferon alfacon 1 or consensus interferon (CIFN) given with RBV, was evaluated in patients who failed initial PEG-IFN/RBV therapy. The intent-to-treat analysis included 487 patients; 245 received CIFN 9 microg/day and RBV, and 242 received CIFN 15 microg/day and RBV. Within this group of patients, 59.3% had documented advanced fibrosis at baseline liver biopsy (stage F3 or F4). SVR rates were 6.9% (17/245 patients) in the 9 microg group and 10.7% (26/242) in the 15 microg group. In the intent-to-treat analysis, SVR rates were higher among patients with a >2-log(10) decrease in hepatitis C virus RNA during prior PEG-IFN/RBV therapy: 11% (4/38) in the 9 mug group and 23% (7/31) in the 15 microg group. Among patients with lower baseline fibrosis scores (F0-F3), SVR rates were 7.8% (15/192) in the 9 microg group and 13.1% (23/175) in the 15 microg group. In this same group of patients (F0-F3), if a >2-log(10) decrease in hepatitis C virus RNA with previous PEG IFN/RBV treatment was achieved, SVR rates improved to 10.7% and 31.6% in the 9 microg and 15 microg groups, respectively. CIFN/RBV combination retreatment was safe and well tolerated. CONCLUSION: Retreatment of PEG-IFN and RBV nonresponders with CIFN and RBV is safe and efficacious and can be considered a retreatment strategy for patients failing previous therapy with PEG-IFN/RBV, especially in interferon-sensitive patients with lower baseline fibrosis scores. PMID- 19291791 TI - Processed allografts and type I collagen conduits for repair of peripheral nerve gaps. AB - Autografting is the gold standard in the repair of peripheral nerve injuries that are not amenable to end-to-end coaptation. However, because autografts result in donor-site defects and are a limited resource, an effective substitute would be valuable. In a rat model, we compared isografts with Integra NeuraGen (NG) nerve guides, which are a commercially available type I collagen conduit, with processed rat allografts comparable to AxoGen's Avance human decellularized allograft product. In a 14-mm sciatic nerve gap model, isograft was superior to processed allograft, which was in turn superior to NG conduit at 6 weeks postoperatively (P < 0.05 for number of myelinated fibers both at midgraft and distal to the graft). At 12 weeks, these differences were no longer apparent. In a 28-mm graft model, isografts again performed better than processed allografts at both 6 and 22 weeks; regeneration through the NG conduit was often insufficient for analysis in this long graft model. Functional tests confirmed the superiority of isografts, although processed allografts permitted successful reinnervation of distal targets not seen in the NG conduit groups. Processed allografts were inherently non-immunogenic and maintained some internal laminin structure. We conclude that, particularly in a long gap model, nerve graft alternatives fail to confer the regenerative advantages of an isograft. However, AxoGen processed allografts are superior to a currently available conduit-style nerve guide, the Integra NeuraGen. They provide an alternative for reconstruction of short nerve gaps where a conduit might otherwise be used. PMID- 19291792 TI - CD133+ and nestin+ tumor-initiating cells dominate in N29 and N32 experimental gliomas. AB - The current study was designed to critically evaluate the notion that cancer stem cell (CSC)-like cells constitute a subpopulation of cells within experimental gliomas. Virtually all cells within the N29 and N32 rat glioma models homogenously expressed CD133, the stem/progenitor marker nestin as well as the neural lineage markers glial fibrillary acidic protein, betaIII-tubulin, and CNPase in vitro. The phenotype was largely retained on exposure to conditions promoting differentiation in vitro and after intracranial implantation of tumor cells into syngeneic hosts. Unsorted adherently grown cells displayed very high clonogenicity in vitro and robust tumorigenicity in vivo. Single N29 and N32 tumor cells invariably formed clones in vitro, and intracerebral inoculation of as few as 10 adherently growing N29 and N32 tumor cells, respectively, gave rise to a tumor. These results provide an alternative view on CSC-like cells in glioma models: sphere-formation is not a prerequisite for accumulation of tumorigenic cells, and CSC-like cells do not reside within a rare subpopulation of cells in these glioma models. N29 and N32 gliomas may accordingly be used for the development of treatment strategies directed specifically against a practically pure population of brain tumor-initiating CSC-like cells. PMID- 19291793 TI - Pml and TAp73 interacting at nuclear body mediate imatinib-induced p53 independent apoptosis of chronic myeloid leukemia cells. AB - Bcr-abl signals for leukemogenesis of chronic myeloid leukemia (CML) and activates ras. Since the function of promyelocytic leukemia protein (pml) is provoked by ras to promote apoptosis and senescence in untransformed cells, the function is probably masked in CML. Imatinib specifically inhibits bcr-abl and induces apoptosis of CML cells. As reported previously, p53(wild) CML was more resistant to imatinib than that lacking p53. Here, we searched for an imatinib induced p53 independent proapoptotic mechanism. We found imatinib up-regulated phosphorylation of p38 mitogen-activated protein kinase (MAPK), checkpoint kinase 2 (chk2) and transactivation-competent (TA) p73; expression of pml and bax; formation of PML-nuclear body (NB); and co-localization of TAp73/PML-NB in p53 nonfunctioning K562 and p53(mutant) Meg-01 CML cells, but not in BCR-ABL(-) HL60 cells. In K562 cells, with short interfering RNAs (siRNAs), knockdown of pml led to dephosphorylation of TAp73. Knockdown of either pml or TAp73 abolished the imatinib-induced apoptosis. Inhibition of p38 MAPK with SB203580 led to dephosphorylation of TAp73, abolishment of TAp73/PML-NB co-localization, and the subsequent apoptosis. Conversely, interferon alpha-2a (IFNalpha), which increased phosphrylated TAp73 and TAp73/PML-NB co-localization, increased additively apoptosis with imatinib. The imatinib-induced TAp73/PML-NB co-localization was accompanied by co-immpunoprecipitation of TAp73 with pml. The imatinib-induced co localization was also found in primary CML cells from 3 of 6 patients, including 2 with p53(mutant) and one with p53(wild). A novel p53-independent proapoptotic mechanism using p38 MAPK /pml/TAp73 axis with a step processing at PML-NB and probably with chk2 and bax being involved is hereby evident in some imatinib treated CML cells. PMID- 19291794 TI - Localization of nuclear cathepsin L and its association with disease progression and poor outcome in colorectal cancer. AB - Previous in vitro studies have identified a nuclear isoform of Cathepsin L. The aim of this study was to examine if nuclear Cathepsin L exists in vivo and examine its association with clinical, pathological and patient outcome data. Cellular localization (nuclear and cytoplasmic) and expression levels v of Cathespin L in 186 colorectal cancer cases using immunohistochemistry. The molecular weight and activity of nuclear and cytoplasmic Cathepsin L in vivo and in vitro were assessed by Western blotting and ELISA, respectively. Epithelial nuclear staining percentage (p = 0.04) and intensity (p = 0.006) increased with advancing tumor stage, whereas stromal cytoplasmic staining decreased (p = 0.02). Using multivariate statistical analysis, survival was inversely associated with staining intensity in the epithelial cytoplasm (p = 0.01) and stromal nuclei (p = 0.007). In different colorectal cell lines and in vivo tumors, pro- and active Cathepsin L isoforms were present in both the cytoplasm and nuclear samples, with pro-Cathepsin L at 50 kDa and active Cathepsin L at 25 kDa. Purified nuclear and cytoplasmic fractions from cell lines and tumors showed active Cathepsin L activity. The identification of nuclear Cathepsin L may play an important prognostic role in colorectal disease progression and patient outcome. Moreover, these findings suggest that altering active nuclear Cathepsin L may significantly influence disease progression. PMID- 19291795 TI - Proteomic and phosphoproteomic alterations in benign, premalignant and tumor human breast epithelial cells and xenograft lesions: biomarkers of progression. AB - The MCF10A human breast epithelial cell lineage includes the benign MCF10A cells, premalignant cells (MCF10AT, MCF10ATG3B) and malignant MCF10CA1a tumor cells. The premalignant and tumor cells recapitulate the progressive alterations associated with the temporal development of PBD and carcinoma. Ras protein levels were elevated by 6.9-, 22.4- and 32.2-fold in 10AT, 10ATG3B and 10CA1a cells, respectively, relative to 10A cells. K-Ras was not detected, N-Ras levels were unchanged; Rac and Rho levels increased in 10CA1a tumor cells. Phospho phosphatidylinositol 3-kinase, phosphoinositide-dependent protein kinase 1 (PDK1), phospho-PDK1, phospho-eukaryotic translation initiation factor 4E (eIF4E) and phospho-eukaryotic initiation factor 4E binding protein 1 (4E-BP1) levels progressively increased in the cell lineage, with the greatest increase monitored in 10CA1a tumor cells. Phospho Ser 473 and Thr 408 Akt levels increased 10.2- and 136-fold in 10CA1a cells, respectively, relative to 10A cells. Phospho-p70S6 kinase (p70S6K) increased >2-fold in 10CA1a cells, relative to 10A cells. Immunohistochemistry confirmed Ras, phospho-Akt and phospho-p70S6K (Thr 421/ Ser 424) expression in lesions arising from premalignant and tumor cells. FOXO 1, phospho-FOXO 1 and phospho-FOXO 4 were significantly elevated in 10ATG3B premalignant and 10CA1a tumor cells. Phospho-FOXO 3a was progressively elevated, with the greatest levels detected in 10CA1a tumor cells. Immunohistochemistry revealed that phospho-FOXO 1, 3a and 4 staining was less in benign lesions, but elevated in advanced 10ATG3B and malignant 10CA1a lesions, showing a correspondence between the cells and lesions. Hence, phospho-Akt and phospho-FOXO 1, 3a and 4 merit consideration as biomarkers of tumorigenic risk from hyperplastic breast tissue. PMID- 19291796 TI - Survivin minigene DNA vaccination is effective against neuroblastoma. AB - The inhibitor of apoptosis protein survivin is highly expressed in neuroblastoma (NB) and survivin-specific T cells were identified in Stage 4 patients. Therefore, we generated a novel survivin minigene DNA vaccine (pUS-high) encoding exclusively for survivin-derived peptides with superior MHC class I (H2-K(k)) binding affinities and tested its efficacy to suppress tumor growth and metastases in a syngeneic NB mouse model. Vaccination was performed by oral gavage of attenuated Salmonella typhimurium SL7207 carrying pUS-high. Mice receiving the pUS-high in the prophylactic setting presented a 48-52% reduction in s.c. tumor volume, weight and liver metastasis level in contrast to empty vector controls. This response was as effective as a survivin full-length vaccine and was associated with an increased target cell lysis, increased presence of CD8(+) T-cells at the primary tumor site and enhanced production of proinflammatory cytokines by systemic CD8(+) T cells. Furthermore, depletion of CD8(+) but not CD4(+) T-cells completely abrogated the pUS-high mediated primary tumor growth suppression, demonstrating a CD8(+) T-cell mediated effect. Therapeutic vaccination with pUS-high led to complete NB eradication in over 50% of immunized mice and surviving mice showed an over 80% reduction in primary tumor growth upon rechallenge in contrast to controls. In summary, survivin-based DNA vaccination is effective against NB and the rational minigene design provides a promising approach to circumvent potentially hazardous effects of using full length antiapoptotic genes as DNA vaccines. PMID- 19291797 TI - Haemochromatosis HFE gene polymorphisms as potential modifiers of hereditary nonpolyposis colorectal cancer risk and onset age. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is characterized by germline mutations in DNA mismatch repair genes; however, variation in disease expression suggests that there are potential modifying factors. Polymorphisms of the HFE gene, which cause the iron overload disorder hereditary haemochromatosis, have been proposed as potential risk factors for the development of colorectal cancer (CRC). To understand the relationship between HNPCC disease phenotype and polymorphisms of the HFE gene, a total of 362 individuals from Australia and Poland with confirmed causative MMR gene mutations were genotyped for the HFE C282Y and H63D polymorphisms. A significantly increased risk of developing CRC was observed for H63D homozygotes when compared with combined wild-type homozygotes and heterozygotes (hazard ratio = 2.93, p = 0.007). Evidence for earlier CRC onset was also observed in H63D homozygotes with a median age of onset 6 years earlier than wild type or heterozygous participants (44 vs. 50 years of age). This effect was significant by all tests used (log-rank test p = 0.026, Wilcoxon p = 0.044, Tarone-Ware p = 0.035). No association was identified for heterozygosity of either polymorphism and limitations on power-prevented investigation of C282Y homozygosity or compound C282Y/H63D heterozygosity. In the Australian sample only, women had a significantly reduced risk of developing CRC when compared with men (hazard ratio = 0.58, p = 0.012) independent of HFE genotype for either single nucleotide polymorphisms. In conclusion, homozygosity for the HFE H63D polymorphism seems to be a genetic modifier of disease expression in HNPCC. Understanding the mechanisms by which HFE interrelates with colorectal malignancies could lead to reduction of disease risk in HNPCC. PMID- 19291798 TI - Anatomical information is needed in ultrasound imaging of muscle to avoid potentially substantial errors in measurement of muscle geometry. AB - This study validates two-dimensional (2D) ultrasound measurements of muscle geometry of the human medial gastrocnemius (GM) and investigates effects of probe orientation on errors in these measurements. Ultrasound scans of GM muscle belly were made both on human cadavers (n = 4) and on subjects in vivo (n = 5). For half of the cadavers, ultrasound scans obtained according to commonly applied criteria of probe orientation deviated 15 degrees from the true fascicle plane. This resulted in errors of fascicle length and fascicle angle up to 14% and 23%, respectively. Fascicle-like structures were detectable over a wide range of probe tilt and rotation angles, but they did not always represent true fascicles. Errors of measurement were either linear or quadratic functions of tilt angle. Similar results were found in vivo. Therefore, we conclude that similar errors are likely to occur for in vivo measurements. For all cadavers, at the distal end of GM, the true fascicle plane was shown to be perpendicular to the distal aponeurosis. Using transverse images of GM to detect the curvature of the deep aponeurosis at the distal end of the muscle belly is a simple strategy to help identify the fascicle plane. For subsequent longitudinal imaging, probe alignment within this plane will help minimize measurement errors of fascicle length, fascicle angle, and muscle thickness. Muscle Nerve, 2009. PMID- 19291799 TI - Fast-twitch sarcomeric and glycolytic enzyme protein loss in inclusion body myositis. AB - Inclusion body myositis (IBM) is an inflammatory disease of skeletal muscle of unknown cause. To further understand the nature of the tissue injury in this disease, we developed methods for large-scale detection and quantitation of proteins in muscle biopsy samples and analyzed proteomic data produced by these methods together with histochemical, immunohistochemical, and microarray data. Twenty muscle biopsy samples from patients with inflammatory myopathies (n = 17) or elderly subjects without neuromuscular disease (n = 3) were profiled by proteomic studies using liquid chromatographic separation of peptides followed by mass spectrometry. Thirteen of the diseased samples additionally underwent microarray studies. Seventy muscle specimens from patients with a range of neuromuscular disorders were examined by ATPase histochemical methods. Smaller numbers of samples underwent immunohistochemical and immunoblot studies. Mass spectrometric studies identified and quantified approximately 300 total distinct proteins in each muscle sample. In IBM and to a lesser extent in polymyositis, proteomic studies confirmed by histochemical, immunohistochemical, and immunoblot studies showed loss of many fast-twitch specific structural proteins and glycolytic enzymes despite relative preservation of transcript levels. Increased abundance of a nuclear membrane protein, immunoglobulins, and two calpain-3 substrates were present. The atrophy present in IBM muscle is accompanied by preferential loss of fast-twitch structural proteins and glycolytic enzymes, particularly glycogen debranching enzyme, with relative preservation of the abundance of their respective transcripts. Although muscle atrophy has long been recognized in IBM, these studies are the first to report specific proteins which are reduced in quantity in IBM muscle. PMID- 19291800 TI - Characterization of circulating tumor cells by fluorescence in situ hybridization. AB - Tumor cells in blood of patients with metastatic carcinomas have been associated with poor survival prospects. Further characterization of these cells may provide further insights into the metastatic process. Circulating Tumor Cells (CTC) were enumerated in 7.5 mL of blood with the CellSearch system. After enumeration of Cytokeratin+, CD45-, nucleated cells, the cells are fixed in the cartridge while maintaining their original position. Cartridges were hybridized with FISH probes against the centromeric regions of chromosome 1, 7, 8, and 17. Next fluorescence images of the FISH probes of the previous identified CTC were acquired. Leukocytes surrounding the CTC were used as internal controls. The number of copies of chromosome 1, 7, 8, and 17 could be determined in 118 CTC containing blood samples from 59 metastatic prostate cancer patients. The samples contained a total of 21,751 CTC (mean 184, median 16, SD 650). Chromosome counts were obtained in 61% of the relocated CTC. On an average, these CTC contained 2.8 copies of chromosome 1, 2.7 copies of chromosome 7, 3.1 copies of chromosome 8, and 2.3 copies of chromosome 17. CTC in which no chromosome count was obtained most likely underwent apoptosis indicated by the expression of M30. In 6/59 patients only diploid CTC were detected these samples, however, only contained 1 5 CTC. Heterogeneity in the chromosomal abnormalities was observed between CTC of different patients as well as among CTC of the same patient. Cytogenetic composition of CTC can be reliably assessed after they have been identified by the CellSearch system. The majority of CTC in hormone refractory prostate cancer are aneuploid confirming that they indeed are cancer cells. An extensive heterogeneity in the copy number of each of the chromosomes was observed. PMID- 19291801 TI - HLA-DRneg patients without acute promyelocytic leukemia show distinct immunophenotypic, genetic, molecular, and cytomorphologic characteristics compared to acute promyelocytic leukemia. AB - BACKGROUND: Loss of HLA-DR and CD34 is a well-known characteristic of malignant promyelocytes in acute promyelocytic leukemia (APL). However, this immunophenotype is not specific for APL. The purpose of this study was to investigate whether further biological characterization of the HLA-DR(neg) acute myeloid leukemia patients would allow more clearly define criteria to separate APL from non-APL patients. METHODS: Immunophenotyping, cytogenetics, molecular analyses, and cytomorphology were prospectively performed within routine leukemia diagnostics of 800 patients included in different prospective acute myeloid leukemia multicenter trials. RESULTS: Beside 60 APL, an additional 62 HLA-DR(neg) non-APL patients were identified. The main differential characteristics of HLA DR(neg) non-APL included high CXCR-4 expression in most patients and almost all leukemia cells, a significantly higher proportion of patients presenting with NPM1 mutations as well as the significant association with cup-like nuclear morphology. The biological distinctness of both leukemia subtypes was further emphasized by the complete absence of aberrant CD2 expression and increased leukocyte and platelet counts in HLA-DR(neg) non-APL patients. Even in the CD34(pos) subgroup of HLA-DR(neg) non-APL all those features contributed in at least the same way to the separation from APL. CONCLUSIONS: The results of the present study show that an immunophenotypic, molecular, and cytomorphologic separation of both HLA-DR(neg) leukemia subgroups is possible indicating that both groups are biologically distinct. PMID- 19291802 TI - Short-term lime pretreatment of poplar wood. AB - Short-term lime pretreatment uses lime and high-pressure oxygen to significantly increase the digestibility of poplar wood. When the treated poplar wood was enzymatically hydrolyzed, glucan and xylan were converted to glucose and xylose, respectively. To calculate product yields from raw biomass, these sugars were expressed as equivalent glucan and xylan. To recommend pretreatment conditions, the single criterion was the maximum overall glucan and xylan yields using a cellulase loading of 15 FPU/g glucan in raw biomass. On this basis, the recommended conditions for short-term lime pretreatment of poplar wood follow: (1) 2 h, 140 degrees C, 21.7 bar absolute and (2) 2 h, 160 degrees C, and 14.8 bar absolute. In these two cases, the reactivity was nearly identical, thus the selected condition depends on the economic trade off between pressure and temperature. Considering glucose and xylose and their oligomers produced during 72 h of enzymatic hydrolysis, the overall yields attained under these recommended conditions follow: (1) 95.5 g glucan/100 g of glucan in raw biomass and 73.1 g xylan/100 g xylan in raw biomass and (2) 94.2 g glucan/100 g glucan in raw biomass and 73.2 g xylan/100 g xylan in raw biomass. The yields improved by increasing the enzyme loading. An optimal enzyme cocktail was identified as 67% cellulase, 12% beta-glucosidase, and 24% xylanase (mass of protein basis) with cellulase activity of 15 FPU/g glucan in raw biomass and total enzyme loading of 51 mg protein/g glucan in raw biomass. Ball milling the lime-treated poplar wood allowed for 100% conversion of glucan in 120 h with a cellulase loading of only 10 FPU/g glucan in raw biomass. PMID- 19291803 TI - Separation of product associating E. coli host cell proteins OppA and DppA from recombinant apolipoprotein A-I(Milano) in an industrial HIC unit operation. AB - We have shown how product associating E. coli host cell proteins (HCPs) OppA and DppA can be substantially separated from apolipoprotein A-I(Milano) (apo A-I(M)) using Butyl Sepharose hydrophobic interaction chromatography (HIC). This work illustrates the complex problems that frequently arise during development and scale-up of biopharmaceutical manufacturing processes. Product association of the HCPs is confirmed using co-immunoprecipitation and Western blotting techniques. Two-dimensional gel electrophoresis and mass spectrometry techniques are used to confirm the identity of OppA and DppA. In this example, clearance of these difficult to separate HCPs decreased significantly when the process was scaled to a 1.4 m diameter column. Laboratory-scale experimentation and trouble shooting identified several key parameters that could be further optimized to improve HCP clearance. The key parameters included resin loading, peak cut point on the ascending side, wash volume, and wash salt concentration. By implementing all of the process improvements that were identified, it was possible to obtain adequate HCP clearance so as to meet the final specification. Although it remains speculative, it is believed that viscosity effects may have contributed to the lower HCP clearance observed early in the manufacturing campaign. PMID- 19291804 TI - Kinetics of CHO A L mutant expression after treatment with gamma radiation, EMS, and asbestos. AB - The flow cytometry mutation assay (FCMA) uses hybrid CHO A(L) cells to measure mutations of the cd59 gene located on human chromosome 11 by the absence of fluorochrome-conjugated antibody binding to the CD59 surface antigen. Mutant expression peaks between 6 and 12 days, then decreases to a stable plateau, instead of a constant mutant fraction obtained by clonogenic assays. To evaluate this variable mutant expression time, cells were treated with radiation, EMS or asbestos and cell proliferation and survival were measured at times leading up to peak mutant expression. Potential doubling time (T(pot)) values increased by at least 75% for each agent by 3 h after treatment but returned to control levels after only 3 days. Survival returned to 90% of control within a week, close to the peak expression day for all three agents. The survival of CD59(-) cells sorted on the peak day of expression was roughly half that of CD59(+) cells. Cloned EMS-treated CD59(-) cells had a doubling time of 16.7 vs. 14.1 h for CD59(+) cells. Triple mutants (CD59(-)/CD44(-)/CD90(-)) were preferentially lost from the population over time, while the proportion of CD59(-)/CD90(-) mutants increased. In conclusion, the peak day of mutant expression occurs only when cells recover from the toxic effects of the mutagen. A fraction of cells originally quantified as mutants are lost over time due to lethal deletions and slower growth. PMID- 19291805 TI - Generation of digital phantoms of cell nuclei and simulation of image formation in 3D image cytometry. AB - Image cytometry still faces the problem of the quality of cell image analysis results. Degradations caused by cell preparation, optics, and electronics considerably affect most 2D and 3D cell image data acquired using optical microscopy. That is why image processing algorithms applied to these data typically offer imprecise and unreliable results. As the ground truth for given image data is not available in most experiments, the outputs of different image analysis methods can be neither verified nor compared to each other. Some papers solve this problem partially with estimates of ground truth by experts in the field (biologists or physicians). However, in many cases, such a ground truth estimate is very subjective and strongly varies between different experts. To overcome these difficulties, we have created a toolbox that can generate 3D digital phantoms of specific cellular components along with their corresponding images degraded by specific optics and electronics. The user can then apply image analysis methods to such simulated image data. The analysis results (such as segmentation or measurement results) can be compared with ground truth derived from input object digital phantoms (or measurements on them). In this way, image analysis methods can be compared with each other and their quality (based on the difference from ground truth) can be computed. We have also evaluated the plausibility of the synthetic images, measured by their similarity to real image data. We have tested several similarity criteria such as visual comparison, intensity histograms, central moments, frequency analysis, entropy, and 3D Haralick features. The results indicate a high degree of similarity between real and simulated image data. PMID- 19291806 TI - Relationship between ultra-weak bioluminescence and vigour or irradiation dose of irradiated wheat. AB - Ultra-weak luminescent analysis is a new way to detect the irradiation dose and the vigour of irradiated wheat. Wheat grain and wheat flour were used in this research for ultra-weak luminescent analysis. The experimental data showed that the bioluminescence intensity of wheat grain sample was different with increasing storage time and increasing dose, and a similar trend appeared in the germination rates of irradiated wheat grain. It was found that the differences in bioluminescence intensities and germination rates of irradiated wheat grain at different doses and storage times were due to the effect of irraditation on the wheat embryo and self-repair during storage. As a result, ultra-weak luminescent analysis cannot be used to detect the irradiation dose of irradiated wheat, but it can be used to determine vigour. Experiments showed that the irradiation dose had a highly significant effect on the bioluminescence intensities of wheat flour when cane sugar was added. PMID- 19291807 TI - A novel chemiluminescent flow injection analysis of trace amounts of rutin by its inhibition of the luminol-hydrogen peroxide reaction catalyzed by tetrasulfonated colbalt phthalocyanine. AB - Based on the inhibition effect of rutin on the luminol-hydrogen peroxide chemiluminescence (CL) system catalyzed by tetrasulfonated colbalt phthalocyanine (CoTSPc), a sensitive flow-injection CL method has been developed for the determination of rutin. The CL reaction mechanism was carefully investigated by examining CL emission spectra, UV-visible spectra and variation of reaction conditions. It was found that there existed a linear relationship between CL intensity and the concentration of rutin in the range of 8.0 x 10(-9) to 1.0 x 10(-6) mol L(-1), and the detection limit is 3.8 x 10(-9) mol L(-1). This proposed method is sensitive, convenient and simple, and has been applied to the determination of rutin in commercial rutin tablets with satisfactory results. PMID- 19291808 TI - Analytical quantification of low-density lipoprotein using europium tetracycline indicator. AB - Low-density lipoprotein (LDL) is known as 'bad' cholesterol. If too much LDL circulates in the blood it can be retained in the walls of the arteries, causing atherosclerosis. In this paper we showed an alternative method to quantify LDL using the europium tetracycline (EuTc) indicator. The optical properties of the EuTc complex were investigated in aqueous solutions containing LDL. An enhancement was observed of the europium luminescence in the solutions with LDL compared those without the lipoprotein. A method to quantify the amount of LDL in a sample, based on EuTc enhanced luminescence, is proposed. The enhancement mechanism is also discussed. PMID- 19291809 TI - Catalytic resonance scattering spectral determination of ultratrace horseradish peroxidase using rhodamine S. AB - A highly sensitive and selective resonance scattering spectral assay was proposed for the determination of horseradish peroxidase (HRP), based on its catalytic effect on the H2O2 oxidation of KI to form I3(-). The I3(-) combined respectively with rhodamine (Rh) dye such as rhodamine S (RhS), rhodamine 6G (Rh6G), rhodamine B (RhB) and butyl-rhodamine B (b-RhB), to form association particles (Rh-I3(n). The four Rh systems all exhibit a stronger resonance scattering (RS) peak at 424 nm. For the RhS, Rh6G, RhB and b-RhB systems, HRP concentration in the range of 3.2 x 10(-12) to 4.8 x 10(-9), 2 x 10(-11) to 3.2 x 10(-9), 1.6 x 10(-11) to 3.2 x 10(-9) and 1.6 x 10(-11) to 4 x 10(-9) g/mL was linear to its RS intensity at 424 nm, with a detection limit of 2.2 x 10(-12), 2.5 x 10(-12), 4.4 x 10(-12) and 2.6 x 10(-12 )g/mL, respectively. This RhS system was most sensitive and stable, and was applied for the determination of HRP in the hepatitis B surface antibody labeling HRP and water samples, with satisfactory results. PMID- 19291810 TI - Luminol-dependent chemiluminescence of human phagocyte cell lines: comparison between DMSO differentiated PLB 985 and HL 60 cells. AB - The human promyelocytic leukemia HL 60 and PLB 985 cell lines can differentiate into terminally mature neutrophil-like cells via dimethyl sulfoxide (DMSO) induction. In this study the luminol-dependent chemiluminescence (LCL) of both neutrophil-like cells was analayzed and compared in response to phorbol myristate acetate (PMA) and opsonized zymosan (OZ) stimulants. It was shown that, like human blood neutrophils, both neutrophil-like cells expressed high levels of CD11b, but unlike human blood neutrophils these cells almost lack LCL-detectable intracellular oxidase activity. By studying the pattern of activation to OZ and PMA and priming with GM-CSF, we concluded that there is no difference between the percentage of differentiation and function of DMSO-induced HL 60 and PLB 985. However, the LCL capacity (area under the curve) of DMSO induced PLB 985 cells was higher than that of HL 60 cells in response to both PMA and OZ, which implies a higher capacity to generate reactive oxygen species in PLB 985 cells. PMID- 19291811 TI - A bioluminescent enzyme immunoassay for prostaglandin E(2) using Cypridina luciferase. AB - Prostaglandin E(2) is one of the major cyclooxygenase metabolites of arachidonic acid. We developed a competitive immunosorbent assay for prostaglandin E(2) utilizing a bioluminescent enzyme Cypridina luciferase. The prostaglandin E(2) amount could be quantified over the concentration ranging from 7.8 to 500 pg/mL. The amount of unlabeled prostaglandin E(2) required to displace 50% of the maximal binding of Cypridina luciferase-labeled prostaglandin E(2) (B/B(0)) was approximately 35 pg/mL. The results show a great potential of Cypridina luciferase as a new labeling enzyme for enzyme-linked immunosorbent assay. PMID- 19291812 TI - Influence of Tl+ activator ions on the luminescence characteristics of KCl0.5Br0.5:Eu2+ powder phosphors. AB - Photoluminescence (PL) of thallium co-doped with KCl0.5Br0.5:Eu2+ powder phosphors display emission bands at 320 and 370 nm attributable to centres involving Tl+ ions in addition to characteristic Eu2+ emission around 420 nm. Additional PL excitation and emission bandS observed around 260 and 380 nm, respectively, were observed in the double-doped KCl0.5Br0.5:Eu2+, Tl+ powder phosphors and are attributed to complex centres involving Tl+ and Eu2+ ions. The enhancement observed in the intensity of Eu2+ emission around 420 nm with the addition of TlBr in KCl0.5Br0.5:Eu2+ powder phosphors is attributed to the energy transfer from Tl+ --> Eu2+ ions. Photostimulated luminescence (PSL) studies of gamma-irradiated KCl0.5Br0.5:Eu2+, Tl+ mixed phosphors are reported and a tentative PSL mechanism in the phosphors has been suggested. PMID- 19291813 TI - Safety of ultrasonography in pregnancy: WHO systematic review of the literature and meta-analysis. AB - OBJECTIVE: In the context of the planned International Society of Ultrasound in Obstetrics and Gynecology-World Health Organization multicenter study for the development of fetal growth standards for international application, we conducted a systematic review and meta-analysis to evaluate the safety of human exposure to ultrasonography in pregnancy. METHODS: A systematic search of electronic databases, reference lists and unpublished literature was conducted for trials and observational studies that assessed short- and long-term effects of exposure to ultrasonography, involving women and their fetuses exposed to ultrasonography, using B-mode or Doppler sonography during any period of pregnancy, for any number of times. The outcome measures were: (1) adverse maternal outcome; (2) adverse perinatal outcome; (3) abnormal childhood growth and neurological development; (4) non-right handedness; (5) childhood malignancy; and (6) intellectual performance and mental disease. RESULTS: The electronic search identified 6716 citations, and 19 were identified from secondary sources. A total of 61 publications reporting data from 41 different studies were included: 16 controlled trials, 13 cohort and 12 case-control studies. Ultrasonography in pregnancy was not associated with adverse maternal or perinatal outcome, impaired physical or neurological development, increased risk for malignancy in childhood, subnormal intellectual performance or mental diseases. According to the available clinical trials, there was a weak association between exposure to ultrasonography and non-right handedness in boys (odds ratio 1.26; 95% CI, 1.03-1.54). CONCLUSION: According to the available evidence, exposure to diagnostic ultrasonography during pregnancy appears to be safe. PMID- 19291814 TI - Islet transplantation and antioxidant management: a comprehensive review. AB - Islet transplantation as a promising treatment for type 1 diabetes has received widespread attention. Oxidative stress plays an essential role in cell injury during islet isolation and transplantation procedures. Antioxidants have been used in various studies to improve islet transplantation procedures. The present study reviews the role of oxidative stress and the benefits of antioxidants in islet transplantation procedures. The bibliographical databases Pubmed and Scopus were searched up to November 2008. All relevant human and animal in-vivo and in vitro studies, which investigated antioxidants on islets, were included. Almost all the tested antioxidants used in the in-vitro studies enhanced islet viability and insulin secretion. Better control of blood glucose after transplantation was the major outcome of antioxidant therapy in all in-vivo studies. The data also indicated that antioxidants improved islet transplantation procedures. Although there is still insufficient evidence to draw definitive conclusions about the efficacy of individual supplements, the benefits of antioxidants in islet isolation procedures cannot be ignored. PMID- 19291815 TI - Nuclear effects of ethanol-induced proteasome inhibition in liver cells. AB - Alcohol ingestion causes alteration in several cellular mechanisms, and leads to inflammation, apoptosis, immunological response defects, and fibrosis. These phenomena are associated with significant changes in the epigenetic mechanisms, and subsequently, to liver cell memory. The ubiquitin-proteasome pathway is one of the vital pathways in the cell that becomes dysfunctional as a result of chronic ethanol consumption. Inhibition of the proteasome activity in the nucleus causes changes in the turnover of transcriptional factors, histone modifying enzymes, and therefore, affects epigenetic mechanisms. Alcohol consumption has been associated with an increase in histone acetylation and a decrease in histone methylation, which leads to gene expression changes. DNA and histone modifications that result from ethanol-induced proteasome inhibition are key players in regulating gene expression, especially genes involved in the cell cycle, immunological responses, and metabolism of ethanol. The present review highlights the consequences of ethanol-induced proteasome inhibition in the nucleus of liver cells that are chronically exposed to ethanol. PMID- 19291816 TI - In vitro and in vivo models of acute alcohol exposure. AB - Alcohol abuse is a global problem due to the financial burden on society and the healthcare system. While the harmful health effects of chronic alcohol abuse are well established, more recent data suggest that acute alcohol consumption also affects human wellbeing. Thus, there is a need for research models in order to fully understand the effect of acute alcohol abuse on different body systems and organs. The present manuscript summarizes the interdisciplinary advantages and disadvantages of currently available human and non-human models of acute alcohol abuse, and identifies their suitability for biomedical research. PMID- 19291817 TI - Autophagy and ethanol-induced liver injury. AB - The majority of ethanol metabolism occurs in the liver. Consequently, this organ sustains the greatest damage from ethanol abuse. Ethanol consumption disturbs the delicate balance of protein homeostasis in the liver, causing intracellular protein accumulation due to a disruption of hepatic protein catabolism. Evidence indicates that ethanol or its metabolism impairs trafficking events in the liver, including the process of macroautophagy, which is the engulfment and degradation of cytoplasmic constituents by the lysosomal system. Autophagy is an essential, ongoing cellular process that is highly regulated by nutrients, endocrine factors and signaling pathways. A great number of the genes and gene products that govern the autophagic response have been characterized and the major metabolic and signaling pathways that activate or suppress autophagy have been identified. This review describes the process of autophagy, its regulation and the possible mechanisms by which ethanol disrupts the process of autophagic degradation. The implications of autophagic suppression are discussed in relation to the pathogenesis of alcohol-induced liver injury. PMID- 19291818 TI - Is the iron regulatory hormone hepcidin a risk factor for alcoholic liver disease? AB - Despite heavy consumption over a long period of time, only a small number of alcoholics develop alcoholic liver disease. This alludes to the possibility that other factors, besides alcohol, may be involved in the progression of the disease. Over the years, many such factors have indeed been identified, including iron. Despite being crucial for various important biological processes, iron can also be harmful due to its ability to catalyze Fenton chemistry. Alcohol and iron have been shown to interact synergistically to cause liver injury. Iron-mediated cell signaling has been reported to be involved in the pathogenesis of experimental alcoholic liver disease. Hepcidin is an iron-regulatory hormone synthesized by the liver, which plays a pivotal role in iron homeostasis. Both acute and chronic alcohol exposure suppress hepcidin expression in the liver. The sera of patients with alcoholic liver disease, particularly those exhibiting higher serum iron indices, have also been reported to display reduced prohepcidin levels. Alcohol-mediated oxidative stress is involved in the inhibition of hepcidin promoter activity and transcription in the liver. This in turn leads to an increase in intestinal iron transport and liver iron storage. Hepcidin is expressed primarily in hepatocytes. It is noteworthy that both hepatocytes and Kupffer cells are involved in the progression of alcoholic liver disease. However, the activation of Kupffer cells and TNF-alpha signaling has been reported not to be involved in the down-regulation of hepcidin expression by alcohol in the liver. Alcohol acts within the parenchymal cells of the liver to suppress the synthesis of hepcidin. Due to its crucial role in the regulation of body iron stores, hepcidin may act as a secondary risk factor in the progression of alcoholic liver disease. The clarification of the mechanisms by which alcohol disrupts iron homeostasis will allow for further understanding of the pathogenesis of alcoholic liver disease. PMID- 19291819 TI - Impact of asialoglycoprotein receptor deficiency on the development of liver injury. AB - The asialoglycoprotein (ASGP) receptor is a well-characterized hepatic receptor that is recycled via the common cellular process of receptor-mediated endocytosis (RME). The RME process plays an integral part in the proper trafficking and routing of receptors and ligands in the healthy cell. Thus, the mis-sorting or altered transport of proteins during RME is thought to play a role in several diseases associated with hepatocyte and liver dysfunction. Previously, we examined in detail alterations that occur in hepatocellular RME and associated receptor functions as a result of one particular liver injury, alcoholic liver disease (ALD). The studies revealed profound ethanol-mediated impairments to the ASGP receptor and the RME process, indicating the importance of this receptor and the maintenance of proper endocytic events in normal tissue. To further clarify these observations, studies were performed utilizing knockout mice (lacking a functional ASGP receptor) to which were administered several liver toxicants. In addition to alcohol, we examined the effects following administration of anti-Fas (CD95) antibody, carbon tetrachloride (CCl(4)) and lipopolysaccharide (LPS)/galactosamine. The results of these studies demonstrated that the knockout mice sustained enhanced liver injury in response to all of the treatments, as shown by increased indices of liver damage, such as enhancement of serum enzyme levels, histopathological scores, as well as hepatocellular death. Overall, the work completed to date suggests a possible link between hepatic receptors and liver injury. In particular, adequate function and content of the ASGP receptor may provide protection against various toxin-mediated liver diseases. PMID- 19291820 TI - Hepatitis C virus and ethanol alter antigen presentation in liver cells. AB - Alcoholic patients have a high incidence of hepatitis C virus (HCV) infection. Alcohol consumption enhances the severity of the HCV disease course and worsens the outcome of chronic hepatitis C. The accumulation of virally infected cells in the liver is related to the HCV-induced inability of the immune system to recognize infected cells and to develop the immune responses. This review covers the effects of HCV proteins and ethanol on major histocompatibility complex (MHC) class I- and class II-restricted antigen presentation. Here, we discuss the liver which functions as an immune privilege organ; factors, which affect cleavage and loading of antigenic peptides onto MHC class I and class II in hepatocytes and dendritic cells, and the modulating effects of ethanol and HCV on antigen presentation by liver cells. Altered antigen presentation in the liver limits the ability of the immune system to clear HCV and infected cells and contributes to disease progression. HCV by itself affects dendritic cell function, switching their cytokine profile to the suppressive phenotype of interleukin-10 (IL-10) and transforming growth factor beta (TGFbeta) predominance, preventing cell maturation and allostimulation capacity. The synergistic action of ethanol with HCV results in the suppression of MHC class II-restricted antigen presentation. In addition, ethanol metabolism and HCV proteins reduce proteasome function and interferon signaling, thereby suppressing the generation of peptides for MHC class I-restricted antigen presentation. Collectively, ethanol exposure further impairs antigen presentation in HCV-infected liver cells, which may provide a partial explanation for exacerbations and the poor outcome of HCV infection in alcoholics. PMID- 19291821 TI - Alcohol metabolites and lipopolysaccharide: roles in the development and/or progression of alcoholic liver disease. AB - The onset of alcoholic liver disease (ALD) is initiated by different cell types in the liver and a number of different factors including: products derived from ethanol-induced inflammation, ethanol metabolites, and the indirect reactions from those metabolites. Ethanol oxidation results in the production of metabolites that have been shown to bind and form protein adducts, and to increase inflammatory, fibrotic and cirrhotic responses. Lipopolysaccharide (LPS) has many deleterious effects and plays a significant role in a number of disease processes by increasing inflammatory cytokine release. In ALD, LPS is thought to be derived from a breakdown in the intestinal wall enabling LPS from resident gut bacterial cell walls to leak into the blood stream. The ability of adducts and LPS to independently stimulate the various cells of the liver provides for a two hit mechanism by which various biological responses are induced and result in liver injury. Therefore, the purpose of this article is to evaluate the effects of a two-hit combination of ethanol metabolites and LPS on the cells of the liver to increase inflammation and fibrosis, and play a role in the development and/or progression of ALD. PMID- 19291822 TI - Alcohol-induced protein hyperacetylation: mechanisms and consequences. AB - Although the clinical manifestations of alcoholic liver disease are well described, little is known about the molecular basis of liver injury. Recent studies have indicated that ethanol exposure induces global protein hyperacetylation. This reversible, post-translational modification on the epsilon amino groups of lysine residues has been shown to modulate multiple, diverse cellular processes ranging from transcriptional activation to microtubule stability. Thus, alcohol-induced protein hyperacetylation likely leads to major physiological consequences that contribute to alcohol-induced hepatotoxicity. Lysine acetylation is controlled by the activities of two opposing enzymes, histone acetyltransferases and histone deacetylases. Currently, efforts are aimed at determining which enzymes are responsible for the increased acetylation of specific substrates. However, the greater challenge will be to determine the physiological ramifications of protein hyperacetylation and how they might contribute to the progression of liver disease. In this review, we will first list and discuss the proteins known to be hyperacetylated in the presence of ethanol. We will then describe what is known about the mechanisms leading to increased protein acetylation and how hyperacetylation may perturb hepatic function. PMID- 19291823 TI - Role of cathepsin B-mediated apoptosis in fulminant hepatic failure in mice. AB - AIM: To investigate the pathogenic role of cathepsin B and the protective effect of a cathepsin B inhibitor (CA-074Me) in fulminant hepatic failure in mice. METHODS: LPS/D-Gal N was injected into mice of the model group to induce fulminant hepatic failure; the protected group was administered CA-074me for 30 min before LPS/D-Gal N treatment; the normal group was given isochoric physiologic saline. Liver tissue histopathology was determined with HE at 2, 4, 6 and 8 h after Lps/D-Gal injection. Hepatocyte apoptosis was examined by TUNEL method. The expression of cathepsin B in liver tissues was investigated by immunohistochemistry, Western blot and RT-PCR. RESULTS: Compared with the normal group, massive typical hepatocyte apoptosis occurred in the model group; the number of apoptotic cells reached a maximum 6 h after injection. The apoptosis index (AI) in the protected group was clearly reduced (30.4 +/- 2.8 vs 18.1 +/- 2.0, P < 0.01 ). Cathepsin B activity was markedly increased in drug-treated mice compared with the normal group (P < 0.01). Incubation with LPS/D-Gal N at selected time points resulted in a time-dependent increase in cathepsin B activity, and reached a maximum by 8 h. The expression of cathepsin B was significantly decreased in the protected group (P < 0.01). CONCLUSION: Cathepsin B plays an essential role in the pathogenesis of fulminant hepatic failure, and the cathepsin B inhibitor CA-074me can attenuate apoptosis and liver injury. PMID- 19291824 TI - Clinical significance of "anti-HBc alone" in human immunodeficiency virus positive patients. AB - AIM: To determine the prevalence and clinical relevance of isolated antibodies to hepatitis B core antigen as the only marker of infection ("anti-HBc alone") among human immunodeficiency virus (HIV) type-1 infected patients. Occult hepatitis B infection frequency was also evaluated. METHODS: Three hundred and forty eight histories from 2388 HIV-positive patients were randomly reviewed. Patients with serological markers of hepatitis B virus (HBV) infection were classified into three groups: past hepatitis, "anti-HBc alone" and chronic hepatitis. Determination of DNA from HBV, and RNA and genotype from hepatitis C virus (HCV) were performed on "anti-HBc alone" patients. RESULTS: One hundred and eighty seven (53.7%) HIV-positive patients had markers of HBV infection: 118 past infection (63.1%), 14 chronic hepatitis (7.5%) and 55 "anti-HBc alone" (29.4%). Younger age [2.3-fold higher per every 10 years younger; 95% confidence intervals (CI) 1.33-4.00] and antibodies to HCV infection [odds ratio (OR) 2.87; 95% CI 1.10-7.48] were factors independently associated with the "anti-HBc alone" pattern. No differences in liver disease frequency were detected between both groups. Serum levels of anti-HBs were not associated with HCV infection (nor viral replication or HCV genotype), or with HIV replication or CD4 level. No "anti-HBc alone" patient tested positive for HBV DNA. CONCLUSION: "Anti-HBc alone" prevalence in HIV-positive patients was similar to previously reported data and was associated with a younger age and with antibodies to HCV infection. In clinical practice, HBV DNA determination should be performed only in those patients with clinical or analytical signs of liver injury. PMID- 19291825 TI - Prognostic factors for 5-year survival after local excision of rectal cancer. AB - AIM: To evaluate the prognostic factors for 5-year survival after local excision of rectal cancer, and to examine the therapeutic efficacy and surgical indications for this procedure. METHODS: Clinical data, obtained from 106 local rectal cancer excisions performed between January 1980 and December 2005, were retrospectively analyzed. Survival analysis was performed using the Kaplan-Meier method, statistical comparisons were performed using the log-rank test, and multivariate analysis was performed using the Cox proportional hazards model. RESULTS: Transanal, transsacral, and transvaginal excisions were performed in 92, 12, and 2 cases, respectively. The rate of complication, local recurrence, and 5 year survival was 6.6%, 17.0%, and 86.7%, respectively. Univariate analysis showed that T stage, vascular invasion, and local recurrence were related to the prognosis of the cases (P < 0.05). Multivariate analysis showed that T stage [P = 0.011, 95% confidence interval (CI) = 1.194-3.878] and local recurrence (P = 0.022, 95% CI = 1.194-10.160) were the major prognostic factors for 5-year survival of cases after local excision of rectal cancer. CONCLUSION: Local rectal cancer excision is associated with few complications, and suitable for stages Tis and T1 rectal cancer. Prevention of local recurrence, active postoperative follow up, and administration of salvage therapy are the effective methods to increase the efficacy of local excision of rectal cancer. PMID- 19291826 TI - Morphology and ontogeny of dendritic cells in rats at different development periods. AB - AIM: To study the morphology and ontogeny of dendritic cells of Peyer's patches in rats at different development periods. METHODS: The morphometric and flow cytometric analyses were performed to detect all the parameters of villous-crypts axis and the number of OX62(+)DC, OX62(+)CD4(+)SIRP(+)DC, and OX62(+)CD4(-)SIRP( )DC in the small intestine in different groups of rats. The relationship between the parameters of villous-axis and the number of DC and DC subtype were analyzed. RESULTS: All morphometric parameters changed significantly with the development of pups in the different age groups (F = 10.751, 12.374, 16.527, 5.291, 3.486; P = 0.000, 0.000, 0.000, 0.001, 0.015). Villous height levels were unstable and increased from 115.24 microm to 140.43 microm as early as 3 wk postpartum. Villous area increased significantly between 5 and 7 wk postpartum, peeked up to 13817.60 microm(2) at 7 wk postpartum. Villous height and crypt depth ratios were relatively stable and increased significantly from 2.80 +/- 1.01 to 4.54 +/- 1.56, 9-11 wk postpartum. The expression of OX62(+)DC increased from 33.30% +/- 5.80% to 80% +/- 17.30%, 3-11 wk postpartum (F = 5.536, P = 0.0013). OX62(+)CD4(+)SIRP(+)DC subset levels detected in single-cell suspensions of rat total Peyer's patch dendritic cells (PP-DCs) increased significantly from 30.73% +/- 5.16% to 35.50% +/- 4.08%, 5-7 wk postpartum and from 34.20% +/- 1.35% to 43.60% +/- 2.07% 9-11 wk postpartum (F = 7.216, P = 0.005). CONCLUSION: This study confirms the age-related changes in villous-crypt axis differentiation in the small intestine. Simultaneously, there are also development and maturation in rat PP-DCs phenotypic expression. Furthermore, the morphological changes of intestinal mucosa and the development of immune cells (especially DC) peaked at 9 11 wk postpartum, indicating that the intestinal mucosae reached a relatively mature state at 11 wk postpartum. PMID- 19291827 TI - Meta-analysis of capsule endoscopy in patients diagnosed or suspected with esophageal varices. AB - AIM: To review the literature on capsule endoscopy (CE) for detecting esophageal varices using conventional esophagogastroduodenoscopy (EGD) as the standard. METHODS: A strict literature search of studies comparing the yield of CE and EGD in patients diagnosed or suspected as having esophageal varices was conducted by both computer search and manual search. Data were extracted to estimate the pooled diagnostic sensitivity and specificity. RESULTS: There were seven studies appropriate for meta-analysis in our study, involving 446 patients. The pooled sensitivity and specificity of CE for detecting esophageal varices were 85.8% and 80.5%, respectively. In subgroup analysis, the pooled sensitivity and specificity were 82.7% and 54.8% in screened patients, and 87.3% and 84.7% in the screened/patients under surveillance, respectively. CONCLUSION: CE appears to have acceptable sensitivity and specificity in detecting esophageal varices. However, data are insufficient to determine the accurate diagnostic value of CE in the screen/surveillance of patients alone. PMID- 19291828 TI - Antisense expression of PKCalpha improved sensitivity of SGC7901/VCR cells to doxorubicin. AB - AIM: To explore whether antisense blocking of protein kinase C alpha (PKCalpha) would reverse multi-drug resistance (MDR) in the vincristine (VCR)-resistant human gastric cancer cell line SGC7901/VCR. METHODS: SGC7901/VCR cells expressing antisense PKCalpha, SGC7901/VCR/aPKC, were established by transfection with a recombinant plasmid reversely inserted with PKCalpha cDNA. Empty vector (PCI-neo) transfected cell clones, SGC7901/VCR/neo, served as the control. Western blot method was used to detect PKCalpha content in SGC7901, SGC7901/VCR, SGC7901/VCR/neo and SGC7901/VCR/aPKC cells, using PKCalpha-specific antibody. The sensitivity of SGC7901, SGC7901/VCR, SGC7901/VCR/neo and SGC7901/VCR/aPKC cells to doxorubicin (DOX) in vitro was determined by MTT assay. The uptake of DOX in these cells was detected with fluorescence spectrophotometer. RESULTS: Western blot analysis showed that the PKCalpha protein level was about 8.7-fold higher in SGC7901/VCR cells than that in SGC7901 cells, whereas the protein expression of PKCalpha was reduced by 78% in SGC7901/VCR/aPKC cells when compared with the SGC7901/VCR cells. SGC7901/VCR/aPKC cells had a 4.2-fold increase in DOX cytotoxicity, accompanied by a 1.7-fold increase of DOX accumulation in comparison with SGC7901/VCR cells. CONCLUSION: PKCalpha positively regulates MDR in SGC7901 cells, and inhibition of PKCalpha can partially attenuate MDR in human gastric cancer cells. PMID- 19291829 TI - Cannabinoid hyperemesis syndrome: clinical diagnosis of an underrecognised manifestation of chronic cannabis abuse. AB - Cannabis is a common drug of abuse that is associated with various long-term and short-term adverse effects. The nature of its association with vomiting after chronic abuse is obscure and is underrecognised by clinicians. In some patients this vomiting can take on a pattern similar to cyclic vomiting syndrome with a peculiar compulsive hot bathing pattern, which relieves intense feelings of nausea and accompanying symptoms. In this case report, we describe a twenty-two year-old-male with a history of chronic cannabis abuse presenting with recurrent vomiting, intense nausea and abdominal pain. In addition, the patient reported that the hot baths improved his symptoms during these episodes. Abstinence from cannabis led to resolution of the vomiting symptoms and abdominal pain. We conclude that in the setting of chronic cannabis abuse, patients presenting with chronic severe nausea and vomiting that can sometimes be accompanied by abdominal pain and compulsive hot bathing behaviour, in the absence of other obvious causes, a diagnosis of cannabinoid hyperemesis syndrome should be considered. PMID- 19291830 TI - Liver cell adenoma showing sequential alteration of radiological findings suggestive of well-differentiated hepatocellular carcinoma. AB - A liver tumor 35 mm in diameter was found incidentally in a 40-year-old woman who had no history of liver diseases or the use of oral contraceptives. Radiological diagnostics showed the typical findings of liver cell adenoma (LCA). Dynamic computed tomography revealed that the tumor showed a homogenous enhancement in the arterial phase and almost the same enhancement as the surrounding liver parenchyma in the delayed phase. The tumor was found to contain fat on magnetic resonance imaging. A benign fat containing liver tumor was suggested. However, radiological findings altered, which caused us to suspect that a well differentiated hepatocellular carcinoma (HCC) containing fat was becoming dedifferentiated. Partial hepatectomy was performed and the pathological findings showed the typical findings of LCA. This case was an extremely rare LCA, which had no background of risk for LCA and developed the sequential alteration of the radiological findings to suspect well-differentiated HCC. PMID- 19291831 TI - Intraductal papillary mucinous neoplasm in chronic calcifying pancreatitis: egg or hen? AB - Intraductal papillary mucinous neoplasm (IPMN) is an increasingly reported entity. Extensive pancreatic calcification is generally thought to be a sign of chronic pancreatitis, but it may occur simultaneously with IPMN leading to diagnostic difficulties. We report a case of a patient initially diagnosed with chronic calcifying pancreatitis who was later shown to have a malignant IPMN. This case illustrates potential pitfalls in the diagnosis of IPMN in the case of extensive pancreatic calcification as well as clues that may lead the clinician to suspecting the diagnosis. The possible mechanisms of the relation between pancreatic calcification and IPMN are also reviewed. PMID- 19291832 TI - Recent findings in the use of intraoperative parathyroid hormone monitoring in parathyroid disease. AB - PURPOSE OF REVIEW: To discuss recent findings and controversies regarding intraoperative parathyroid hormone monitoring (IPM) in guiding parathyroidectomy. RECENT FINDINGS: IPM is being frequently used in guiding surgeons to complete excision of abnormal glands during parathyroidectomy for sporadic primary hyperparathyroidism (SPHPT). This adjunct is now being used in many centers around the world and has become a standard of care in the treatment of SPHPT. As the use of this technique developed, the understanding of what was necessary to return patients with hyperparathyroidism to a eucalcemic state, namely, excision of all parathyroid tissue secreting high amount of parathyroid hormone, was recognized. Two major controversies have developed during the evolution of IPM guided parathyroidectomy. One is that gland excision based on this modality may not recognize all abnormal glands, which, if not excised, will result in operative failure or recurrent hyperparathyroidism. The second disagreement is a technical one and concerns the best intraoperative protocol to be used. SUMMARY: Parathyroidectomy for SPHPT is highly successful regardless of the operative approach used. Despite the controversies summarized in the present review, IPM has been shown to be accurate as an adjunct to guide parathyroidectomy and has changed the operative management of SPHPT. PMID- 19291833 TI - Very late stent thrombosis after discontinuation of clopidogrel therapy. PMID- 19291834 TI - Editor's introduction: a diverse collection of articles in terms of topic, geographic location of the research, and methodology. PMID- 19291835 TI - India asks: 'Can you spare a few days ... and a few catheters, Doctor?'. PMID- 19291836 TI - Intranasal administration of peptide antigens of HIV with mucosal adjuvant CpG ODN coentrapped in microparticles enhances the mucosal and systemic immune responses. AB - The mucosal immune system acts as a first line of defense against infection caused by luminal pathogens. Because HIV is transmitted primarily via mucosal associated tissues, particularly with sexual transmission, understanding antiviral immunity present at these sites is important. As most of the peptide antigens show poor immunogenicity when immunized alone but after incorporating the same peptide antigens along with adjuvant CpG ODN in microparticles has shown enhanced immunogenicity in the murine model. In the present study we have investigated the immunomodulatory effects of two adjuvants, CpG 1826 and CpG 2006 (Class B, Also known as type K) to the four peptide antigens of HIV such as envelope glycoproteins gp41 Leucine Zipper, gp41 fusion domain and gp120-C2 as well as regulatory protein (Nef) in microparticles, exploring nasal route with single immunization schedule. Peptide (s) alone in the microparticles elicited low peptide specific IgG and IgA peak titres in the sera, whereas the inclusion of CpG ODN with peptides in microparticles significantly enhanced peptide specific IgG and IgA peak titres and such responses were sustained for longer durations. Similarly higher SIgA response was achieved in the mucosal washes with CpG encapsulated in microparticles. Such presence of SIgA in washes was further correlated with the presence of secretory component (SC) in the respective washes. Both adjuvants induced excellent peptide specific IgG and IgA immune responses. Thus the overall study highlighted the importance of CpG ODNs as a mucosal adjuvant for weaker peptide antigens and thus can explore for developing peptide based vaccine against HIV. PMID- 19291837 TI - Regulation of arginase/nitric oxide synthesis axis via cytokine balance contributes to the healing action of malabaricone B against indomethacin-induced gastric ulceration in mice. AB - The role of the arginine-metabolism in the healing action of the Myristica malabarica phenol malabaricone B (mal B) and omeprazole against indomethacin induced stomach ulceration in mouse was investigated. Indomethacin (18 mg kg- 1) was found to induce maximum stomach ulceration in Swiss albino mice on the 3rd day of its administration, which was associated with reduced arginase activity (30.8%, P < 0.01), eNOS expression, along with increased iNOS expression, total NOS activity (5.55 folds, P < 0.001), NO generation (2.19 folds, P < 0.001), and ratio of pro-/anti-inflammatory cytokines. Besides providing comparable healing as omeprazole (3 mg kg- 1 x 3 days), mal B (10 mg kg- 1 x 3 days, p. o.) shifted the iNOS/NO axis to the arginase/polyamine axis as revealed from the increased arginase activity (51.6%, P < 0.001), eNOS expression, and reduced iNOS expression, total NOS activity (approximately 75%, P < 0.001), and NO level (50.6%, P < 0.01). These could be attributed to a favourable anti/pro inflammatory cytokines ratio, generated by mal B. The healing by omeprazole was however, not significantly associated with those parameters. PMID- 19291838 TI - Fluorescent indication that nitric oxide formation in NTS neurons is modulated by glutamate and GABA. AB - Nitric oxide (NO) in NTS plays an important role in regulating autonomic function to the cardiovascular system. Using the fluorescent dye DAF-2 DA, we evaluated the NO concentration in NTS. Brainstem slices of rats were loaded with DAF-2 DA, washed, fixed in paraformaldehyde and examined under fluorescent light. In different experimental groups, NTS slices were pre-incubated with 1 mM l-NAME (a non-selective NOS inhibitor), 1 mM d-NAME (an inactive enantiomere of l-NAME), 1 mM kynurenic acid (a non-selective ionotropic receptors antagonist) or 20 microM bicuculline (a selective GABAA receptors antagonist) before and during DAF-2 DA loading. Images were acquired using a confocal microscope and the intensity of fluorescence was quantified in three antero-posterior NTS regions. In addition, slices previously loaded with DAF-2 DA were incubated with NeuN or GFAP antibody. A semi-quantitative analysis of the fluorescence intensity showed that the basal NO concentration was similar in all antero-posterior aspects of the NTS (rostral intermediate, 15.5 +/- 0.8 AU; caudal intermediate, 13.2 +/- 1.4 AU; caudal commissural, 13.8 +/- 1.4 AU, n = 10). In addition, the inhibition of NOS and the antagonism of glutamatergic receptors decreased the NO fluorescence in the NTS. On the other hand, d-NAME did not affect the NO fluorescence and the antagonism of GABAA receptors increased the NO fluorescence in the NTS. It is important to note that the fluorescence for NO was detected mainly in neurons. These data show that the fluorescence observed after NTS loading with DAF-2 DA is a result of NO present in the NTS and support the concept that NTS neurons have basal NO production which is modulated by l-glutamate and GABA. PMID- 19291839 TI - Obama allocates funds for health-care priorities. PMID- 19291840 TI - Stem-cell experts raise concerns about medical tourism. PMID- 19291841 TI - Complicated lessons: Lorenzo Odone and medical miracles. PMID- 19291847 TI - Something for everyone? Industry has mixed reactions to budget proposals. PMID- 19291846 TI - Raising the bar for boards. AB - The pressure to improve quality of care has brought increased oversight of hospital trustees, with a variety of industry players offering plans for voluntary certification or education. The efforts are winning praise. "When you go in, you don't know what fully to expect," says Durwood Dominy, left, a veteran trustee in Georgia. PMID- 19291848 TI - Adjusting budgets. Providers look to stay in the black during recession. PMID- 19291849 TI - Private practice. Recovery act contains added disclosure accounting. PMID- 19291850 TI - 'Safety net is struggling'. Report: even the insured have access problems. PMID- 19291851 TI - Spurned in Texas. Supreme court rejects group's price-fix challenge. PMID- 19291852 TI - Boosting nursing profile. Underpaid faculty causing shortage, group claims. PMID- 19291853 TI - Grappling with suicide. New Washington law poses dilemma for providers. PMID- 19291854 TI - No alms for ACEP. Emergency doc group off-base with funding request to feds. PMID- 19291855 TI - A long road to reform. The commitment to achieving universal coverage is an important step. PMID- 19291856 TI - It can be done. Disclosure, retail investors key to bond sale. PMID- 19291857 TI - The effect of phototherapy on neutrophils. AB - BACKGROUND: The role of phototherapy on neutrophils has not been reviewed previously. This novel and non-invasive therapeutic approach is of particular interest for potential use in the treatment of pathologic processes in dermatology and infectious diseases in which neutrophils are the primary culprit. OBJECTIVES: The primary aim of this study was to systematically review the role of phototherapy on neutrophils. METHOD: Original publications were identified through searches in PubMed, Medline, Ovid, and the Cochrane Library. Search terms used included "phototherapy and neutrophils," "light therapy and neutrophils," and "laser and neutrophils." Studies were selected based on the level of evidence based research. RESULTS: The literature search revealed a total of 22 controlled laboratory studies that evaluated the role of phototherapy on neutrophils. Among the effects of phototherapy noted were increases in: the respiratory burst of neutrophils, apoptosis of polymorphonuclear cells, and plasma NO and iNOS mRNA. Other notable findings include decreased: number of neutrophils in areas of inflammation, ROS production, neutrophil anti-apoptotic factors, and IL-1beta concentration. Studies on PDT demonstrated neutrophilia and resultant decreased tumor growth. CONCLUSION: Evidence indicates that phototherapy has a significant impact on neutrophils, the effect of which varies according to the specific type of phototherapy. These findings have a variety of potential clinical applications including the treatment of various autoimmune conditions, inflammatory diseases, and cancers. PMID- 19291858 TI - Pretreatment with R(+)-verapamil significantly reduces mortality and cytokine expression in murine model of septic shock. AB - It is well known that cytokines play an important role in the pathogenesis of sepsis and septic shock. There is evidence indicating that the membrane transporter, P-glycoprotein (P-gp), may be involved in the release of cytokines, such as IL-2, IL-4 or IFN-gamma. The aim of this study was to assess the influence of P-gp inhibitor, R(+)-verapamil, on cytokine expression in serum and tissues as well as survival rate of mice with LPS-induced septic shock. These effects were compared with the response to treatment with pentoxifylline, lisofylline, and prednisolone administered alone or after pretreatment with R(+) verapamil. When given as a single agent, R(+)-verapamil significantly decreased serum levels of TNF-alpha and IFN-gamma and protected mice from endotoxin lethality. Moreover, it decreased up-regulated by LPS TNF-alpha gene expression in the liver and lungs. Given concomitantly with immunomodulatory compounds, it enhanced their beneficial impact on the survival of mice with septic shock. The highest increase in survival rate was observed in combination with pentoxifylline (7% vs. 67%). The most striking differences observed between saline and R(+) verapamil pretreated animals on combination therapy included down-regulation of TNF-alpha, higher levels of IL-6, and decreased IFN-gamma concentrations. These results suggest that P-gp may be involved in the release of IFN-gamma, and possibly also TNF-alpha, in mice with septic shock. R(+)verapamil improves survival of mice receiving a lethal dose of LPS and significantly potentiates the protective effect of pentoxifylline and prednisolone against LPS-induced lethality, probably as a result of both P-gp inhibition and a synergistic interaction at the gene level. PMID- 19291859 TI - Glycyrrhiza inflata-derived chalcones, Licochalcone A, Licochalcone B and Licochalcone D, inhibit phosphorylation of NF-kappaB p65 in LPS signaling pathway. AB - Licorice root has been used as a traditional medicine for the treatment of gastric ulcer, bronchial asthma and inflammation. Licochalcone A is a major component of Xinjiang licorice, Glycyrrhiza inflata. Previously we showed that Licochalcone A significantly inhibited LPS-induced NF-kappaB transcriptional activation by abrogating the phosphorylation of NF-kappaB p65 at serine 276. Glycyrrhiza inflata contains not only Licochalcone A but also Licochalcone B, Licochalcone C, Licochalcone D, Echinatin and Isoliquiritigenin, harboring the common structure of chalcones. No chalcones had any effect on LPS-induced IkappaB degradation, nuclear translocation and DNA binding activity of NF-kappaB p65; however, we observed that Licochalcone B and Licochalcone D significantly inhibited LPS-induced phosphorylation at serine 276 and transcriptional activation of NF-kappaB, the same as Licochalcone A. Interestingly, we also found that Licochalcone A, Licochalcone B and Licochalcone D effectively inhibited LPS induced activation of PKA, which is required for the phosphorylation of NF-kappaB p65 at serine 276. Consequently, Licochalcone B and Licochalcone D significantly reduced the LPS-induced production of NO, TNFalpha and MCP-1. On the other hand, Licochalcone C, Echinatin and Isoliquitigenin failed to inhibit LPS-induced NF kappaB activation. These findings suggest that the anti-inflammatory effect of Glycyrrhiza inflata is ascribable to the potent inhibition of NF-kappaB by Licochalcone A, Licochalcone B and Licochalcone D. PMID- 19291860 TI - An adaptive theory of increasing mortality with increasing chronological age in populations in the wild. AB - An "increasing mortality with increasing chronological age in populations in the wild" (IMICAW) is a phenomenon shown by many species, and the greater or smaller (or non-existent) IMICAW has an adaptive value, since it reduces the "mean duration of life" (ML). As Leopold (1961) pointed out, a smaller ML brings about a greater spreading velocity, within the species, of any advantageous mutation.However, this is an argument of group selection and is, therefore, inadequate to demonstrate that within a species a C gene causing IMICAW is stable compared with a C' allele not having this effect. The problem may be solved if we consider the inclusive fitness of C with the hypothesis that the dead individuals are replaced by kin individuals. In such a case, even with low values of the coefficient of relationship (Hamilton, 1971) of the substituting individuals, C tends to be stable and favoured by the selective mechanism as compared with C'. When the preferential replacement by kin individuals does not happen and/or when the turnover of generations is swift enough, C is not favoured and hence IMICAW loses its hypothesized adaptive value. In such cases, survival curves must be of type II or III of Pianka's classification (1970). It is discussed if IMICAW might be a consequence of the action of many harmful genes that express themselves tardily in the course of life. PMID- 19291861 TI - Studies on the mechanism of hypoglycemia in a patient with massive intraperitoneal leiomyosarcoma. PMID- 19291862 TI - [Osteonecrosis of the jaw]. PMID- 19291863 TI - [There is no "quick fix"]. PMID- 19291864 TI - [Physical exercise and type 2 diabetes: Is 3 x 10 minutes a day better than 30 minutes?--secondary publication]. AB - We compared the effects on glycaemic control of two different exercise protocols in elderly men with type 2-diabetes; one 30-minute session per day (1 x 30) and three 10-minute sessions per day (3 x 10). Cardiopulmonary fitness increased in both groups. Improvement in glycaemic control was only found in the 3 x 10 group. A likely explanation is that the total energy expenditure during training performed as multiple sessions is higher than the expenditure achieved during a single session per day. PMID- 19291865 TI - [Aliskiren combined with losartan in type 2-diabetes and nephropathy - secondary publication]. AB - We evaluated the renoprotective effects of adding aliskiren to treatment with losartan in hypertensive patients with type 2 diabetes and nephropathy. A total of 599 patients were randomized to six months of treatment with placebo or aliskiren in addition to losartan 100 mg and optimal antihypertensive therapy. The primary outcome was a reduction in the urinary albumin-creatinine ratio. Aliskiren 300 mg daily reduced the mean urinary albumin-creatinine ratio by 20% (p < 0.001) compared with placebo. The number of adverse events was similar between groups. Aliskiren is renoprotective independently of its blood pressure lowering effect. PMID- 19291866 TI - [Treatment of patients in private hospitals II]. PMID- 19291867 TI - The female labour market in English agriculture during the Industrial Revolution: expansion or contraction? PMID- 19291868 TI - [Comment to a press release]. PMID- 19291869 TI - [Electronic chaos]. PMID- 19291870 TI - [Do we have a bad Medical Association?]. PMID- 19291871 TI - Comparative phylogenetic study of Stichotrichia (Alveolata: Ciliophora: Spirotrichea) based on 18S-rDNA sequences. AB - Since molecular phylogenies of stichotrich ciliates started to be published, some remarkable contradictions to morphology-based classifications have been reported, such as the Convergent Evolution of Urostylids and Uroleptids (CEUU) hypothesis, the Halteria paradox, the polyphyly of Oxytricha and of Stichotrichia. We hypothesized the internal phylogeny of 18S-rDNA from 53 morphological species of stichotrichs and their relationships with Hypotrichia and Oligotrichia using parsimony and neighbor-joining methods, including new data from Pseudouroleptus caudatus and Strongylidium pseudocrassum. Competing phylogenetic scenarios were compared using statistical tests, and the results suggest the reconsideration of both CEUU and the position of Halteria among flexible-body oxytrichids. The polyphyly of Oxytricha was not rejected and the monophyly of Stichotrichia was accepted based on parsimony analysis if Pseudoamphisiella is considered an external (discocephalid related) taxon. PMID- 19291872 TI - [DAMP again]. PMID- 19291873 TI - Isolation and characterization of HC1: a novel human DNA repair gene. AB - Nucleotide excision repair (NER) acts on a broad spectrum of large lesions, while base excision repair removes individual modified bases. Although both processes have been well studied in human cells, novel genes involved in these DNA repair pathways have been described. Using a heterologous complementation approach, we identified a fetal human cDNA that complemented two Escherichia coli mutants that are defective in 3-methyl adenine glycosylase and in three endonucleases, all of which are enzymes with important roles in base excision repair. The central cDNA open reading frame complemented NER mutant strains and promoted an increase in survival rate of bacteria exposed to UV light. The corresponding protein was able to restore nucleotide-excision-repair activity when added to a cell extract from Chinese hamster ovary cells deficient in the ERCC1 protein, an enzyme known to promote incision at the 5' end of the lesion during NER. In contrast, that protein was not able to complement XPG Chinese hamster ovary cells deficient in the 3' incision step of NER. These data indicate a new human repair gene, which we named HC1; it is involved in the recognition of two kinds of DNA lesions and it contributes to the 5' DNA incision step in NER. PMID- 19291874 TI - Characterization of mitochondrial genotypes in the foundation herd of the Canchim beef cattle breed. AB - The Canchim (5/8 Charolais + 3/8 Zebu) beef cattle breed was developed at Southeast-Embrapa Cattle to take advantage of hybrid vigor and to combine the higher growth rate and beef quality of Charolais with tropical adaptations of Zebu. The development of three lineages (old, new, and crossbred) has increased its genetic basis. The genotypic origin (Bos taurus or Bos indicus) of the mitochondrial DNA (mtDNA) of the Canchim breed was unknown. We characterized the mtDNA genotype of this founder herd by allele-specific polymerase chain reaction. The 173 founder Zebu females (62 Indubrasil, 3 Guzerat, and 108 Nellore) and their 6749 offspring were identified. The frequency of B. indicus mtDNA ranged from 1.15 to 2.05% among the descendants (n= 6404) of each maternal line with available DNA, and among animals that were alive (n= 689) in December 2007 among the three lineages. Though mtDNA characterization can be used to direct animal selection, the low frequency of B. indicus mtDNA impairs the evaluation of its effects on production traits in these animals. The high prevalence of B. taurus mtDNA in Canchim proves that the founder Zebu females from the Indubrasil, Guzerat and Nellore breeds were obtained from crosses of Zebu sires with local B. taurus dams. PMID- 19291875 TI - Primary open angle glaucoma was not found to be associated with p53 codon 72 polymorphism in a Brazilian cohort. AB - Primary open angle glaucoma (POAG) is the most common type of glaucoma. The p53 codon 72 Arg-Pro (CGC to CCC) polymorphism of exon 4 affects various biological properties; recently, it was reported that this polymorphism affects the ability to induce apoptosis in vitro. Various genotypes have been found to be significantly associated with POAG. We examined the distribution of this polymorphism in 104 unrelated POAG patients and in 58 normal healthy individuals without history of POAG at the Pronto Clinica de Olhos in Goiania, Brazil. The controls were recruited among individuals undergoing ophthalmological examination. Their genomic DNA was analyzed for p53 gene codon 72 polymorphism by polymerase chain reaction. The Arg72 allele was more common than the Pro72 allele in both groups. There was no significant difference in the distribution of the codon 72 polymorphism between groups (p= 0.3311). The genotype distribution in the POAG group was 23.07 Arg homozygote, 75 heterozygote, and 1.93% Pro homozygote, while in the control group it was 31.04 Arg homozygote, 68.96 heterozygote, and 0% Pro homozygote. We concluded that the p53 codon 72 Arg/Pro polymorphism is not associated with glaucoma in Brazilian patients. PMID- 19291876 TI - PedExpert: a computer program for the application of Bayesian networks to human paternity testing. AB - PedExpert is a Windows-based Bayesian network software, especially constructed to solve problems in parentage testing that are complex because of missing genetic information on the alleged father and/or because they involve genetic mutations. PedExpert automates the creation and manipulation of Bayesian networks, implementing algorithms that convert pedigrees and sets of indispensable information (genotypes, allele frequencies, mutation rates) into Bayesian networks. This program has a novel feature that can incorporate information about gene mutations into tables of conditional probabilities of transmission of alleles from the alleged father to the child, without adding new nodes to the network. This permits using the same Bayesian network in different modes, for analysis of cases that include mutations or not. PedExpert is user-friendly and greatly reduces the time of analysis for complex cases of paternity testing, eliminating most sources of logical and operational error. PMID- 19291877 TI - Antioxidant effect of haptoglobin phenotypes against DNA damage induced by hydrogen peroxide in human leukocytes. AB - Human haptoglobin is classified into three major phenotypes: Hp1-1, Hp2-1 and Hp2 2; there are two autosomal alleles Hp*1 and Hp*2, and the Hp*1 allele has two subtypes, Hp*1F and Hp*1S. Haptoglobin acts as an antioxidant, preventing hemoglobin-driven oxidative damage. We used the comet assay to examine oxidative damage to DNA induced by hydrogen peroxide in human leukocytes; we also looked for differences in the antioxidant capacity of haptoglobin subtypes. Haptoglobin genotypes were determined through allele-specific polymerase chain reaction, visualized on a polyacrylamide gel. The Hp1-1 genotype had the least DNA damage; this indicates that Hp alleles differ in their protective effects against oxidative damage. Among Hp*1 alleles, Hp*1F was the most protective. PMID- 19291878 TI - Genetic parameters for pre-weaning traits in Braunvieh cattle. AB - The objective of this study was to estimate genetic parameters for pre-weaning traits of Braunvieh cattle raised under tropical conditions in Brazil. The weight and weight gain parameters were birth weight (BW, N = 9955), weight at 120 days of age (W120, N = 5901), weaning weight at 205 days (WW, N = 6970), weight gain from birth to 205 days (GAIN205, N = 6013), weight gain from birth to 120 days (GAIN120, N = 5135), and weight gain from 120 to 205 days (GAIN85, N = 4482). Variance components were estimated using the animal model with the MTDFREML software. The relationship matrix included 35,188 animals; phenotypic measures were available for 18,688. Direct and maternal heritability increased from birth to weaning, with estimates of 0.23 +/- 0.037, 0.25+/- 0.050, 0.41+/- 0.059 for direct heritability for BW, W120 and WW, respectively, 0.08 +/- 0.012, 0.15 +/- 0.032, 0.22 +/- 0.036 for maternal genetic effects, and 0.18, 0.14 and 0.16 for total heritability estimates. For pre-weaning gains, estimates of heritability were 0.36 +/- 0.059, 0.30+/- 0.059, 0.12 +/- 0.035 for direct genetic effects of the traits GAIN205, GAIN120 and GAIN85, respectively, 0.23 +/- 0.038, 0.17 +/- 0.037, 0.03 +/- 0.029 for estimates of maternal heritability, and 0.12, 0.13, 0.16 for total heritability, respectively. Genetic correlations between weights were greater between measures taken at shorter intervals. This information can be used to optimize the design of programs for genetic improvement of Braunvieh cattle raised under tropical conditions. PMID- 19291879 TI - Increasing mutagenicity of Sao Goncalo Channel waters based on the Allium cepa test. AB - The Sao Goncalo Channel is of great importance to the conservation of local biodiversity; it also is a water supply source of the city of Pelotas, Brazil, and the surrounding region. We examined the mutagenic activity of its waters. The following items were seasonally investigated in Allium cepa root radicular meristem cells: mitotic index, mitotic anomalies, interphase anomalies, and total anomalies. Water samples were collected from four different stations, Lock Dam, Santa Barbara Channel, Pelotas Creek, and Barra do Laranjal. A drinking water negative control was used. For each sampling station, 8000 cells were counted, 2000 of which by repetition. The data were computed on a database (SPSS), and then analyzed by the chi-square test and the Mann-Whitney U-test. In 2005, the channel water provoked a significantly greater number of anomalies than the control water. The number of anomalies increased in 2007. This suggests that there was an increase in toxic substances in the channel over the years. PMID- 19291880 TI - Bovine papillomavirus DNA in milk, blood, urine, semen, and spermatozoa of bovine papillomavirus-infected animals. AB - Papillomavirus infection in bovines is associated with cutaneous papillomatosis on the hide, udders and other epithelial tissues, as well as in oral respiratory, alimentary and urinary tract mucosa. Bovine papillomavirus (BPV) is also considered the etiological agent of esophageal tumors and the malignant bladder tumors that characterize the clinical condition associated with chronic enzootic hematuria. After infective viral DNA was found in cattle blood and BPV1, 2 and 4 DNA in cattle reproductive and embryonic tissues, we looked for and found BPV DNA in blood, milk, urine, seminal fluid, and spermatozoa of BPV-infected animals. Peripheral blood lymphocyte cultures from BPV-infected animals had high rates of chromosome aberrations, including radial rearrangements that signal oncogenic potential and viral interaction with telomeric regions. The finding of BPV DNA in body fluids and tissues other than the epithelium demonstrates co-infection of other tissues or cell types by papillomavirus and shows the potential role of lymphocytes, seminal fluid and spermatozoa in BPV transmission. Our findings reinforce a peremptory need for prophylactic and therapeutic instruments to curtail this disease in bovine livestock. PMID- 19291881 TI - Distinction between plant samples according to allele dosage by semiquantitative polymerase chain reaction. AB - The lack of informativity of samples from heterozygotic individuals is one of the hindrances in the mapping of quantitative trait loci of outbred populations, since it is not normally possible to identify the origin of each allele. One way to include these individuals in analyses would be to genotype their endosperm, considering that a heterozygote (Aa) has AAa or Aaa endosperm, when the female genitor donates the A or a allele, respectively. We used semiquantitative polymerase chain reaction to determine allele dosages in DNA mixtures, by simulating the observed conditions for endospermic tissue. Semiquantitative polymerase chain reaction on agarose gels, along with regression analysis, allowed differentiation of the samples according to the amount of DNA. This type of information will help decrease the number of non-informative individuals in quantitative trait locus mapping of outbred populations, thereby increasing mapping accuracy. PMID- 19291882 TI - The first fences: fencing the colony of New South Wales, 1788-1823. PMID- 19291883 TI - Trouble with farms at the Census Office: an evaluation of farm statistics from the censuses of 1851-1881 in England and Wales. PMID- 19291884 TI - Varietal innovation and the competitiveness of the British cereals sector, 1760 1930. PMID- 19291885 TI - Word and sign in Elizabethan conflicts with the devil. PMID- 19291886 TI - [Pregnancy shortly after bariatric surgery]. AB - Bariatric surgery is increasingly used to treat morbidly obese patients. Fertility in women may be enhanced after these procedures, owing to substantial weight loss and possibly a decreased absorption of oral contraceptives. We report a pregnancy that occurred two months after biliopancreatic diversion with duodenal switch in a 32-year-old woman. She subsequently developed haemolysis, elevated liver enzymes and low platelets count (HELLP) syndrome and had a weight loss of 43 kg (from the bariatric procedure) until the infant was delivered preterm by caesarean section (due to low activity). The infant was small in relation to the gestational age, with a weight of less than 50 % of the expected (780 g at 29.6 weeks). Histological examination demonstrated a small placenta with insufficient spiral artery trophoblast infiltration, possibly caused either by severe preeclampsia or by maternal nutritional deficiencies. Severe metabolic aberrations may complicate pregnancies after malabsorptive bariatric surgery. Patient preparations before weight-loss operations should include information on fertility and birth control in the postoperative period. Protocols for monitoring of patients that become pregnant after bariatric surgery are needed. PMID- 19291887 TI - [Bariatric surgery and pregnancy]. PMID- 19291888 TI - Autism. PMID- 19291889 TI - "A plan for the abolition of slavery": consistently with the interests of all parties concerned (London, 1828). PMID- 19291890 TI - Convergence or divergence? Recent historical writings on the Rape of Nanjing. PMID- 19291891 TI - Measuring partnerships in public health. AB - (1) Community-based organizations, public health departments, and direct service health care providers tend to dominate public health collaboratives. (2) Three dimensions for assessing the value of a partner are power and influence, active involvement, and resources. (3) Two important factors in developing positive and successful public health collaboratives are trust among partners and reciprocity. PMID- 19291892 TI - Domesticity and difference: male breadwinners, working women, and colonial citizenship in the 1945 Nigerian general strike. PMID- 19291893 TI - Feminism, social science, and the meanings of modernity: the debate on the origin of the family in Europe and the United States, 1860-1914. PMID- 19291894 TI - Millenarian slaves? The Santidade de Jaguaripe and slave resistance in the Americas. PMID- 19291895 TI - Money, a substitute for confidence? Vaughan to Keynes and beyond. PMID- 19291896 TI - Global tuberculosis control amid the world economic crisis. PMID- 19291897 TI - Glucose self-monitoring: necessary--or not? PMID- 19291898 TI - Nature, nurture, and memory in a Socialist utopia: delineating the Soviet socio ethnic body in the age of Socialism. PMID- 19291899 TI - Retraction: A method for discriminating a Japanese chicken, the Nagoya breed, using microsatellite markers (Poult Sci. 85:2124-2086). PMID- 19291900 TI - Prisoners of war and torture need more attention. PMID- 19291901 TI - Balkan immigrants. PMID- 19291902 TI - Detainees in Saudi Arabia. PMID- 19291903 TI - ASSESSMENT OF THE NATURAL HISTORY OF ANTERIOR COMMUNICATING ANEURYSMS. PMID- 19291904 TI - Mammalian chromosomes in vitro. XVII. Idiogram of the Chinese hamster. AB - From 67 diploid male Chinese hamster cells in vitro, an idiogram was constructed. The chromosomes can be classified into 4 natural groups: group I, 2 pairs of large submetacentrics (1 and 2); group II, 3 pairs of medium-sized submetacentrics (the sex pair, 4 and 5); group III, 3 pairs of medium-sized subtelocentrics (6,7, and 8); group IV, 3 pairs of small metacentrics (9,10, and 11). A system of subdividing each chromosome into regions is also suggested. PMID- 19291905 TI - [Efficacy and challenge of medical care within industry--from a 22-year experience working on about 10,000 subjects]. PMID- 19291906 TI - [Employee assistance program (EAP)]. PMID- 19291907 TI - 2008 Frechette Awards. PMID- 19291909 TI - [Role of an attending physician in mental health services in industry]. PMID- 19291908 TI - [Cooperation between psychiatrists and industrial health specialists in industrial mental health services]. PMID- 19291910 TI - [Basis and problems related to the approval of work-related illnesses--in view of the current trend in legal cases concerning work-related disabilities]. PMID- 19291911 TI - [Medical problems concerning Minamata disease and the approach by the Japanese Society of Psychiatry and Neurology]. PMID- 19291912 TI - [Determination of brain death in children]. PMID- 19291913 TI - [Pediatric asthma and obesity]. PMID- 19291914 TI - Global standardisation of HbA1c. AB - HbA1c is used for assessing glycaemic control in patients with diabetes. It is also used for treatment goals and as a target for therapeutic intervention. The Direct Control and Complications Trial in the USA showed that HbA1c can be used to predict the risk of complications. Hence, it is important for HbA1c assays to be standardised. The National Glycohemoglobin Standardization Program (NGSP) in the USA was formed in 1996 so that HbA1c results from different laboratories would be comparable to those reported in the DCCT study. There were also HbA1c standardisation programmes in Sweden and Japan. These three standardisation programmes are, in fact, direct comparison methods (DCMs), and yield different HbA1c results. In 1994, the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) established a Working Group on Standardisation of HbA1c. This working group has developed a global HbA1c reference system with very much improved intra-assay and inter-assay coefficients of variation. Recommendations have been made to report HbA1c results as IFCC-HbA1c values in SI units (mmol HbA1c/mol Hb) and NGSP-HbA1c (%) as well as estimated average glucose (eAG), once a tight relationship has been shown to exist between eAG and HbA1c. PMID- 19291915 TI - Experimental production of clinical-grade dendritic cell vaccine for acute myeloid leukemia. AB - Dendritic cells (DC) are professional antigen presenting cells of the immune system. Through the use of DC vaccines (DC after exposure to tumour antigens), cryopreserved in single-use aliquots, an attractive and novel immunotherapeutic strategy is available as an option for treatment. In this paper we describe an in vitro attempt to scale-up production of clinical-grade DC vaccines from leukemic cells. Blast cells of two relapsed AML patients were harvested for DC generation in serum-free culture medium containing clinical-grade cytokines GM-CSF, IL-4 and TNF-alpha. Cells from patient 1 were cultured in a bag and those from patient 2 were cultured in a flask. The numbers of seeding cells were 2.24 x 10(8) and 0.8 x 10(8), respectively. DC yields were 10 x 10(6) and 29.8 x 10(6) cells, giving a conversion rate of 4.7% and 37%, respectively. These DC vaccines were then cryopreserved in approximately one million cells per vial with 20% fresh frozen group AB plasma and 10% DMSO. At 12 months and 21 months post cryopreservation, these DC vaccines were thawed, and their sterility, viability, phenotype and functionality were studied. DC vaccines remained sterile up to 21 months of storage. Viability of the cryopreserved DC in the culture bag and flask was found to be 50% and 70% at 12 months post cryopreservation respectively; and 48% and 67% at 21 months post cryopreservation respectively. These DC vaccines exhibited mature DC surface phenotypic markers of CD83, CD86 and HLA-DR, and negative for haemopoietic markers. Mixed lymphocyte reaction (MLR) study showed functional DC vaccines. These experiments demonstrated that it is possible to produce clinical grade DC vaccines in vitro from blast cells of leukemic patients, which could be cryopreserved up to 21 months for use if repeated vaccinations are required in the course of therapy. PMID- 19291916 TI - Performance evaluation of the Arkray Adams HA-8160 HbA1c analyser. AB - BACKGROUND: HbA1c measurement is currently routinely used to predict long term outcome of diabetes, thus playing a fundamental role in the management of diabetes. The relationship between HbA1c value and long term diabetic complications has been established by a randomised control Diabetes Control and Complications Trial (DCCT) which used high performance liquid chromatography (HPLC) as a reference method for HbA1c assay. To ensure that HbA1c results from a variety HbA1c assay methods are similar to the DCCT values, the American Diabetes Association (ADA) recommended that all laboratories should use methods certified by the National Glycohemoglobin Standardization Programme (NGSP) with interassay coefficient variation (CV) of < 5% (ideally < 3%). The International Federation of Clinical Chemistry (IFCC) working group on HbA1c standardisation has set a CV < 2.5% as a criteria for its reference laboratories. OBJECTIVES: To evaluate the performance of Arkray Adams HA-8160 HbA1c analyser which uses a cation exchange HPLC method and its correlation to HbA1c assay on Cobas Integra 800 which is an immunoturbidimetric method. METHODS: For the imprecision study, patient samples and control material of two levels were analysed on HA-8160 analyser 20 times in a single run (within-run imprecision) and twice a day on five consecutive days (between-run imprecision). For the recovery study, two samples each with high and low values were selected and mixed in ratios of 1:3, 1:1 and 3:1, and were analysed by HA-8160. Sixty samples were analysed by both Cobas Integra 800 and HA 8160 for method comparison study. Ten uraemic samples and ten thalassaemic samples were assayed on Cobas Integra 800 and HA 8160 for interference study. RESULTS: Within-run CVs were 0.6% and 0.7% for medium and high value samples respectively, 0.6% and 0.7% for low and high level controls respectively. Between run CVs were 0.5% and 0.4% for medium and high value samples respectively, 0.5% and 0.6% for low and high level controls respectively. The mean recovery was 100.1%. A good correlation between the 2 methods (Adams = 1.00 Cobas - 0.11, r = 0.98) was observed. CONCLUSIONS: The Akray Adams HA-8160 HbA1c analyser performed within the target CV of < 2.5% and showed a good correlation with the Cobas Integra 800. PMID- 19291917 TI - Assessment of P-gp and MRP1 activities using MultiDrugQuant Assay Kit: a preliminary study of correlation between protein expressions and its functional activities in newly diagnosed acute leukaemia patients. AB - Multidrug resistance (MDR) is believed to be responsible for poor response of patients towards chemotherapy particularly patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). The best-characterized resistance mechanism is the one mediated by permeability-glycoprotein (P-gp) encoded by MDR1 gene, which is responsible for drug efflux. We studied P-gp and multidrug resistance-associated protein 1 (MRP1) expression and functional activities in 43 newly diagnosed acute leukemia cases (19 paediatric ALL cases and 24 adult AML cases). The expression and functional activities were examined using flow cytometry and MultiDrugQuant assay kit (involving calcein AM uptake and efflux). P-gp and MRP1 expression and its functional activities were observed in 68.4% of paediatric ALL. In adult AML cases, all cases expressed MRP1 and its functional activities but only 58.3% were positive for P-gp and its functional activities. We were able to show a significant correlation between the expression of the multidrug resistant protein (P-gp and MRP1) and their functional activity in adult AML and paediatric ALL samples. PMID- 19291918 TI - The effect of water extracts of Euphorbia hirta on cartilage degeneration in arthritic rats. AB - The effect of water extracts of Euphorbia hirta on the histological features and expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in the rat articular cartilage was investigated. Arthritis was induced in rats using Freund's Complete Adjuvant containing heat killed M. tuberculosis, and treated with water extracts of E. hirta. Paraffin tissue sections of the arthritic joints were evaluated. The extent of cartilage degeneration was found to be greatest in rats treated with the highest dosage of E. hirta, followed by rats in the untreated group. Rats treated with the intermediary and low dosages of Euphorbia hirta showed improved histology. MMP-13 levels were found to be decreased with decreasing dosages of E. hirta. TIMP-1 levels were found to increase with decreasing dosages of E. hirta. MMP-3 levels fluctuated without any appreciable pattern. Low dosages of E. hirta seem to be beneficial in reducing cartilage degeneration in cases of arthritis. PMID- 19291919 TI - Association of Ki67 with raised transaminases in hepatocellular carcinoma. AB - Transaminase enzymes, alanine (ALT) and aspartate transaminase (AST), have been reported to be raised and implicated to have prognostic value in hepatocellular carcinoma (HCC). Ki67, a marker of cellular proliferative activity, has also been noted to be increased in HCC. A study was conducted at the Department of Pathology, Faculty of Medicine, University of Malaya, Kuala Lumpur to determine the possible association of proliferative activity, as determined by Ki67, with the transaminase enzymes. 31 cases of histologically diagnosed HCC who underwent tumour resection were retrieved from departmental archives. The patients' ages ranged between 40 to 79 years with a mean of 58.3 years. There was a male preponderance with M:F = 2.9:1. Ethnic Chinese formed 83.9% of the cases. 4 microm sections, cut from the formalin-fixed, paraffin-embedded tumour tissue block of each case, were immunohistochemically stained with Ki67 (DAKO monoclonal MIB-1) using the commercial DakoCytomation EnVision+System-HRP kit. The latest ALT and AST levels, assayed within 7 days prior to tumour resection, were retrieved from the patients' case records. 24 (77.4%) HCC demonstrated elevation of either ALT and/or AST. 27 (87.1%) HCC were immunopositive for Ki67. Ki67 immunoexpression was significantly correlated with raised transaminases (p<0.05). Hypothetically, the mechanism by which this phenomenon may occur may simply be release of transaminases due to destruction of hepatocytes by the cancer. Thus rising levels of the transaminases could signal a more rapid growth of the tumour and these routinely performed tests can be of prognostic value in management of HCC patients. PMID- 19291920 TI - Biochemical profiling in two siblings with mitochondrial 2-methylacetoacetyl-CoA thiolase deficiency. AB - We report the biochemical profiling in two siblings with mitochondrial 2 methylacetoacetyl-CoA thiolase deficiency. Organic aciduria typical of this rare inborn error metabolism was found when the elder sibling presented with an episode of severe ketoacidosis at 20 months of age, which consisted of excessive excretion of ketones, tiglylglycine, 2-methyl-3-hydroxybutyrate, and 2 methylacetoacetate. Blood acylcarnitiness profile showed elevation of C5OH carnitine, which represents 2-methyl-3-hydroxybutyrylcarnitine. A similar biochemical profile was identified in the younger sibling during screening although he had only mild clinical symptoms. Both patients reported a favourable outcome on follow-up. PMID- 19291921 TI - Follicular dendritic cell sarcoma of inguinal lymph node--a case report. AB - Follicular dendritic cell sarcomas (FDCS) are rare neoplasms that involve lymph nodes or extranodal sites. They show varied histological features and thus can be mistaken for carcinoma or sarcoma. Correct identification is important for further management. A 43-year-old Indian female presented with a three-month history of progressive swelling at the right inguinal region. It was excised completely and was reported as lymph node with metastatic poorly differentiated carcinoma based on Haematoxylin and eosin (H&E) stain findings. Computerized tomography (CT) scans of thorax, abdomen and pelvis were normal and did not reveal a primary site. Following this, the case was referred to one of the authors. The slides were reviewed and a variety of immunocytochemical markers were done. The tumour cells were negative for epithelial, melanocytic, neural, leucocyte and soft tissue tumour markers. They were immunopositive for CD21, CD35 and negative for CD68. Based on the immunocytochemical findings, a final diagnosis of FDCS was made. This case highlights the histological and immunophenotypical profile of a rare tumour which requires a high index of suspicion for diagnosis. PMID- 19291922 TI - Invasive lobular carcinoma of the breast diagnosed from an ovarian tumour. AB - Invasive lobular carcinoma of breast is well known to be able to metastasise to unusual places, including the gastrointestinal and gynaecological tracts. However, it is very unusual for breast cancer to present in an ovarian Brenner tumour. This case highlights the diagnostic difficulties of patients presenting with unilateral ovarian masses, and the merits of thorough histological assessment of the entire pathological specimen despite the presence of one obvious pathological diagnosis. PMID- 19291923 TI - Metastasis of cervical carcinoma to endometrial polyp: an interesting case report. AB - A metastatic focus of small circumscribed carcinoma in an endometrial polyp is extremely rare. Most of these reported cases have a primary carcinoma of the breast. We report a circumscribed metastatic squamous cell carcinoma in an endometrial polyp. This, to the best of our knowledge, is the first case report of metastasis of cervical carcinoma to a benign endometrial polyp. PMID- 19291924 TI - An unusual cause of haemoptysis and headache: cryptococcosis. AB - Pulmonary cryptococcosis can be clinically silent in non-HIV infected patients but can also present as nodules and masses on the chest radiograph, which can be mistaken for tuberculosis or lung cancer. Common symptoms include fever and cough, and uncommonly haemoptysis. This report illustrates a non-HIV infected patient whose main complaint was haemoptysis and headache. He was diagnosed with pulmonary cryptococcosis from biopsy of an endobronchial mass found on flexible bronchoscopy. Disseminated cryptoccoccal infection should be considered as a differential diagnosis in non-HIV infected patients presenting with haemoptysis and headache. Early recognition and administration of appropriate therapy will improve clinical outcome in these patients. PMID- 19291925 TI - Musculo-skeletal issue. PMID- 19291926 TI - Evaluation of the effect of local corticosteroid injection and anti-inflammatory medication in carpal tunnel syndrome. AB - BACKGROUND AND AIMS: To evaluate the effect of local corticosteroid injection versus non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of carpal tunnel syndrome (CTS), 32 patients were included in this study. METHODS: A prospective, randomised clinical trial, assessing functional findings by Functional Status Scale (FSS), (JTT) Jebsen Taylor Test and electrophysiologic examinations, analysed on initial visit and at the third month after treatment. Group A was treated with betamethasone injection and group B with NSAIDs, both with concomitant use of wrist splints. RESULTS: The mean age of the patients was 40.8 +/- 11.2 (range 21-64) years. There was a significant improvement in FSS in groups A and B at the third month. In JTT, 'writing', 'picking up small common objects' and 'picking up large heavy objects' activities were improved in group A; 'writing' and 'stacking checkers' activities were improved in group B. Statistically significant improvement was observed in peak sensory conduction velocity and distal motor latency in groups A and B. Mixed nerve conduction velocity and compound sensory action potential were improved in group A. CONCLUSIONS: The results showed that neither of the groups demonstrated superior results. We conclude that local steroid injection and NSAIDs with concomitant use of wrist splints may offer patients with CTS variable and effective treatment options for the management of functional scores and nerve conduction parameters. PMID- 19291927 TI - A comparison of rural and urban rheumatoid arthritis populations. AB - INTRODUCTION: There is evidence to suggest that remote populations have poorer clinical outcomes in certain disease processes such as asthma and cancer. This study looks to identify any disparities in the management of patients with rheumatoid arthritis in the context of rurality. METHODS: A retrospective observational study was performed on all 1314 patients with a diagnosis of rheumatoid arthritis who have been under the care of the principal rheumatologist at Raigmore Hospital, Inverness, between the years 1994 and 2004 inclusive. Rurality was defined according to the Scottish Household Survey. Populations were assessed in terms of age; sex; duration of diagnosis; number of years of Disease Modifying AntiRheumatic Drugs (DMARD) therapy, prednisolone use and the number of musculoskeletal practical interventions undertaken (eg joint aspiration or replacement). RESULTS: Two thirds of patients were considered rural dwellers. No significant difference was established between the populations with regards to management. DMARD therapy had been prescribed in 77% of rural patients vs 70% of their city counterparts for a mean 5.4 and 4.0 years respectively. The proportion of patients exposed to prednisolone therapy and who underwent musculoskeletal procedures were equivalent. CONCLUSIONS: Rural dwellers, with rheumatoid arthritis in the Highlands of Scotland, do not appear to be disadvantaged in regards to their disease management in comparison to the urban population. PMID- 19291928 TI - Direct to consumer advertising via the Internet, a study of hip resurfacing. AB - BACKGROUND AND AIMS: With increased use of the internet for health information and direct to consumer advertising from medical companies, there is concern about the quality of information available to patients. The aim of this study was to examine the quality of health information on the internet for hip resurfacing. METHODS: An assessment tool was designed to measure quality of information. Websites were measured on credibility of source; usability; currentness of the information; content relevance; content accuracy/completeness and disclosure/bias. Each website assessed was given a total score, based on number of scores achieved from the above categories websites were further analysed on author, geographical origin and possession of an independent credibility check. RESULTS: There was positive correlation between the overall score for the website and the score of each website in each assessment category. Websites by implant companies, doctors and hospitals scored poorly. Websites with an independent credibility check such as Health on the Net (HoN) scored twice the total scores of websites without. CONCLUSIONS: Like other internet health websites, the quality of information on hip resurfacing websites is variable. This study highlights methods by which to assess the quality of health information on the internet and advocates that patients should look for a statement of an "independent credibility check" when searching for information on hip resurfacing. PMID- 19291929 TI - Use of radiology by general practitioners making orthopaedic referrals: a cross sectional study. AB - BACKGROUND: In orthopaedic clinics, patients can be x-rayed during their visit to the surgeon. Could radiographs carried out in primary care be more efficiently carried out when the patient meets the surgeon? AIMS: To quantify and describe use of radiological investigations by general practitioners (GPs), when referring to orthopaedics. METHODS: We made a retrospective examination of 294 referral letters for detail regarding x-ray investigation of the condition prompting the letter. We used referral letters assessed by the recipient as 'routine'. We excluded those addressed to a particular surgeon because of subspecialty interest or prior care. RESULTS: Twenty one percent (62) of referrals described an investigation that did not contribute to management prior to the patient's clinic appointment. CONCLUSIONS: A significant number of patients travelled specifically for an x-ray. In some of these cases the radiographs could be delayed until the clinic visit without compromising care. DISCUSSION: We suggest that GPs seeing patients with musculoskeletal problems should consider referral to an orthopaedic surgeon prior to requesting x-rays. X-ray investigation in the orthopaedic clinic may be more convenient for the patient and surgeon. PMID- 19291930 TI - Displaced intracapuslar hip fractures in the working age alcohol-abusing patient. AB - Management of alcohol-abusing patients presenting with intracapsular hip fractures is controversial. The aim of this study was to compare the management and outcome of working-age alcohol-abusing patients with similar-aged controls. Patients were identified from a prospectively collected database of trauma admissions. Full case notes were available for 78 patients under 65 of age presenting with a displaced intacapsular fracture at a teaching hospital between 1998 and 2002. Thirty seven patients had evidence of alcohol abuse. Alcohol abusing patients presented to hospital later (p = 0.05), underwent surgery a median of 18 hours later (p = 0.011) and required a longer post operative stay (p = 0.003) compared to non-abusers. Despite this, the results of internal fixation were comparable. There was no significant difference between alcohol-abusers and non-abusers in rates of avascular necrosis (6.9% vs 9.7%; odds ratio 0.69, 0.11 4.47) or revision surgery (0.21 vs 0.10 procedures/ patient; odds ratio 1.49, 0.30-7.33). The high rates of alcohol abuse in this low-velocity trauma population suggest such patients are at increased risk of osteoporosis. Routine screening for osteoporosis should be considered in working-age alcohol abusers. After subcapital fracture, reduction and internal fixation is an acceptable treatment in this sub-group of patients. PMID- 19291931 TI - Hormone replacement therapy (HRT) or etidronate for osteoporosis in postmenopausal asthmatics on glucocorticoids: a randomised factorial trial. AB - INTRODUCTION: The study was designed to establish the effects of HRT on osteoporosis and fractures over five years in postmenopausal women with asthma receiving regular glucocorticoids and to compare with etidronate. METHODS: Postmenopausal patients receiving inhaled and/or oral glucocorticoids were randomly assigned to HRT, cyclical etidronate, HRT plus cyclical etidronate or no treatment for five years. The trial was multi-centre and aimed to recruit 750 patients. Outcomes were fractures and changes in bone mineral density (BMD). RESULTS: For reasons detailed in the discussion section of the text, only 50 patients were entered. Three did not fulfil the eligibility criteria and were excluded from the analysis. Among the remaining 47 patients, three (6%) experienced new, symptomatic fractures, one on etidronate and two in the no treatment group. New or worsening morphometric fractures of the thoracolumbar spine occurred in 50% of the 22 patients with spinal radiographs on entry and at five years (one HRT, three etidronate, two HRT plus etidronate and five on no treatment). BMD improved by approximately 1% per annum in those receiving HRT and/or etidronate; comparisons of HRT vs no HRT tended to favour HRT but were only statistically significant at proximal femur. The same trends emerged in the etidronate vs no etidronate comparison, but none reached the 5% level of statistical significance. DISCUSSION: For postmenopausal patients receiving glucocorticoids for asthma, HRT appears as effective as etidronate in preventing loss of BMD over five years and may have a similar effect on fracture prevention. PMID- 19291932 TI - Weight changes following lower limb arthroplasty: a prospective observational study. AB - The aim of this study was to assess patterns of weight loss/gain following total hip or knee joint replacement. Four hundred and fifty primary lower limb arthroplasty patients, where the current surgery was the last limiting factor to improved mobility, were selected. Over a one year period 212 gained weight (mean 5.03kg), 92 remained static, and 146 lost weight. The median change was a weight gain of 0.50Kg (p = 0.002). All patients had a significant improvement in Oxford outcome scores. Hip arthroplasty patients were statistically more likely to gain weight than knee arthroplasty patients. A successful arthroplasty, restoring a patient's mobility, does not necessarily lead to subsequent weight loss. The majority of patients put on weight with an overall net weight gain. No adverse effect on functional outcome was noted. PMID- 19291933 TI - Manipulation under anaesthesia and early physiotherapy facilitate recovery of patients with frozen shoulder syndrome. AB - AIM: The aim of this study was to examine the efficacy of manipulation under anaesthesia (MUA) followed by early physiotherapy in treating frozen shoulder syndrome. METHODS: In a prospective trial conducted between 26th August 2002 and 25th June 2004 in 86 patients with frozen shoulder syndrome, MUA was performed as a day procedure. Main outcomes were Disabilities of the Arm, Shoulder and Hand (DASH) score, a visual analogue score (VAS) for pain and range of movement, which were measured preoperatively and at six weeks postprocedure. RESULTS: Fifty (58.1%) patients had complete data. The average age was 54.5 years. The mean duration of symptoms until MUA was 13 months. The mean DASH score decreased from 48.07 to 15.84 (p < 0.0005). The mean VAS reduced from 6.07 to 1.88 (p < 0.0005). Flexion improved from 104.18 to 157.56 (p < 0.0005); abduction from 70.48 to 150.00 (p < 0.0005); and external rotation from 13.88 to 45.62 (p < 0.0005). CONCLUSION: MUA combined with early physiotherapy alleviates pain and facilitates recovery of function in patients with frozen shoulder syndrome. PMID- 19291934 TI - An observational study of the impact of a rehabilitation admission on readmission data. AB - BACKGROUND AND AIM: Specialist psychiatric rehabilitation services are changing in response to overall redesign of psychiatric services. The assumed therapeutic gains of these services have not been established or quantified, and their role in contemporary practice has not been formally articulated. The aim of this observational study is to examine the impact of an admission to psychiatric rehabilitation wards at the Royal Edinburgh Hospital on readmission data post discharge. METHOD: Thirty-five admissions were included in the sample. Simple measures of outcome were used: the number of days spent in hospital; number of Mental Health Act uses; number of admissions. This information was obtained from case notes and from data collected routinely on the Patient Information Management System. These measures were examined in the two-year periods before and after a rehabilitation admission. RESULTS: In the two-year period following the index admission there was a significant reduction in all these measures compared to the two-year period before the index admission: mean number of bed days (p < 0.001); number of uses of the Mental Health Act (p < 0.001); number of readmissions (p < 0.01). CONCLUSION: There was an improvement in patient outcome measures following a rehabilitation admission. PMID- 19291935 TI - NICE guidance in the Scottish context. AB - The UK landscape of guidance for clinical practice and public health has become increasingly complicated in recent times, and it is easy to feel confused as to what is available for what purposes, what applies where, and who does what. Within this overall picture, there is potential for uncertainty as to the applicability, or otherwise, in Scotland of guidance produced by the National Institute for Health and Clinical Excellence (NICE) in England, including NICE technology appraisals, clinical guidelines, interventional procedures guidance, and public health programme and intervention guidance. This paper is offered as a map to help health professionals and others to find their way through the Scottish landscape, with particular reference to NICE's outputs and the respective roles of NHS Quality Improvement Scotland (NHS QIS) and NHS Health Scotland (HS). PMID- 19291936 TI - Urogynaecology: a review. PMID- 19291937 TI - Twentieth century orthopaedics: its development as a specialty in Aberdeen. PMID- 19291938 TI - [Determination of procalcitonin. Valuable for reducing ineffective antibiotic use is debatable]. PMID- 19291939 TI - [Eighty-plussers with aorta stenosis. Age itself is no contra-indicator for surgery]. PMID- 19291940 TI - [Angioplasty in stable angina pectoris. Not cost effective compared to drug treatment]. PMID- 19291941 TI - [Assessment clinical and surgical competency. Less agreement among assessors by using 'Short clinical assessment' than with 'Objective structured assessment of technical skills']. PMID- 19291942 TI - [Diagnostic images (406). A neonate with obstructive breathing. Solitary median maxillary central incisor'-syndrome]. PMID- 19291943 TI - [Exceptionally long survival]. PMID- 19291944 TI - [Cancer stem cells]. PMID- 19291945 TI - [Acetylsalicylic acid does not prevent cardiovascular diseases in diabetic patients]. PMID- 19291946 TI - [Postnatal ocular toxoplasmosis in a grown woman]. PMID- 19291947 TI - [Intensive care or not in immature children?]. PMID- 19291948 TI - [A rare cause of clubbing of fingers and watch glass nails]. PMID- 19291949 TI - [The social impact of applied health research. Measure and make visible]. PMID- 19291950 TI - [Obtaining medical ethical approval for a multicentre, randomised study: prospective evaluation of a ponderous process]. PMID- 19291951 TI - [Holistic medicine]. PMID- 19291952 TI - Exposure limits in peril. PMID- 19291953 TI - Sedentary confinement. PMID- 19291955 TI - Release the brakes and fly without fear. PMID- 19291954 TI - Go beyond OSHA compliance. PMID- 19291956 TI - Developing a culture of alertness. PMID- 19291957 TI - Refining safety. PMID- 19291958 TI - Advancing safety around the world. PMID- 19291959 TI - Getting the message. PMID- 19291960 TI - It's time for an annual refresher! PMID- 19291961 TI - Vision testing: a blind spot in occupational safety. PMID- 19291962 TI - Stay on your toes. PMID- 19291963 TI - Determining confined space training requirements. PMID- 19291964 TI - Pinpointing a pathogen. PMID- 19291965 TI - For safety's sake. PMID- 19291966 TI - Special delivery. PMID- 19291967 TI - Point/counterpoint. Preparation for a terrorism-related radiation event should be no different from that for a biological or chemical event. PMID- 19291968 TI - A low-cost density reference phantom for computed tomography. AB - The authors characterized a commercially available foam composed of polyurethane and polyisocyanurate which is marketed for modeling parts in the aircraft, automotive, and related industries. The authors found that the foam may be suitable for use as a density reference standard in the range below -400 Hounsfield units. This range is coincident with the density of lung tissue. The foam may be helpful in making the diagnosis of lung disease more systematic. PMID- 19291969 TI - In vivo 3D modeling of the femoropopliteal artery in human subjects based on x ray angiography: methodology and validation. AB - Endovascular revascularization of the femoropopliteal (FP) artery has been limited by high rates of restenosis and stent fracture. The unique physical forces that are applied to the FP artery during leg movement have been implicated in these phenomena. The foundation for measuring the effects of physical forces on the FP artery in a clinically relevant environment is based on the ability to develop 3D models of this vessel in different leg positions in vivo in patients with peripheral arterial disease (PAD). By acquiring paired angiographic images of the FP artery, and using angiography-based 3D modeling algorithms previously validated in the coronary arteries, the authors generated 3D models of ten FP arteries in nine patients with PAD with the lower extremity in straight leg (SL) and crossed leg (CL) positions. Due to the length of the FP artery, overlapping paired angiographic images of the entire FP artery were required to image the entire vessel, which necessitated the development of a novel fusion process in order to generate a 3D model of the entire FP artery. The methodology of angiographic acquisition and 3D model generation of the FP artery is described. In a subset of patients, a third angiographic view (i.e., validation view) was acquired in addition to the standard paired views for the purpose of validating the 3D modeling process. The mean root-mean-square (rms) error of the point-to point distances between the centerline of the main FP artery from the 2D validation view and the centerline from the 3D model placed in the validation view for the SL and CL positions were 0.93 +/- 0.19 mm and 1.12 +/- 0.25 mm, respectively. Similarly, the mean rms error of the same comparison for the main FP artery and sidebranches for the SL and CL positions were 1.09 +/- 0.38 mm and 1.21 +/- 0.25 mm, respectively. A separate validation of the novel fusion process was performed by comparing the 3D model of the FP artery derived from fusion of 3D models of adjacent FP segments with the 2D validation view incorporating the region of fusion. The mean rms error of vessel centerline points of the main FP artery, the main FP artery plus directly connected sidebranches, and the mean rms error of upstream, downstream, and sidebranch directional vectors at bifurcation points in the overlap region were 1.41 +/- 0.79 mm, 2.13 +/- 1.12 mm, 3.16 +/- 3.72 degrees, 3.60 +/- 5.39 degrees, and 8.68 +/- 8.42 degrees in the SL position, respectively, and 1.29 +/- 0.35 mm, 1.61 +/- 0.78 mm, 4.68 +/- 4.08 degrees, 3.41 +/- 2.23 degrees, and 5.52 +/- 4.41 degrees in the CL position, respectively. Inter- and intraobserver variability in the generation of 3D models of individual FP segments and the fusion of overlapping FP segments were assessed. The mean rms errors between the centerlines of nine 3D models of individual FP segments generated by two independent observers, and repeated measurement by the same observer were 2.78 +/- 1.26 mm and 3.50 +/- 1.15 mm, respectively. The mean rms errors between the centerline of four 3D models of fused overlapping FP segments generated by two independent observers, and repeated measurement by the same observer were 4.99 +/- 0.99 mm and 5.98 +/- 1.22 mm, respectively. This study documents the ability to generate 3D models of the entire FP artery in vivo in patients with PAD in both SL and CL positions using routine angiography, and validates the methodologies used. PMID- 19291970 TI - Computer-aided mass detection in mammography: false positive reduction via gray scale invariant ranklet texture features. AB - In this work, gray-scale invariant ranklet texture features are proposed for false positive reduction (FPR) in computer-aided detection (CAD) of breast masses. Two main considerations are at the basis of this proposal. First, false positive (FP) marks surviving our previous CAD system seem to be characterized by specific texture properties that can be used to discriminate them from masses. Second, our previous CAD system achieves invariance to linear/nonlinear monotonic gray-scale transformations by encoding regions of interest into ranklet images through the ranklet transform, an image transformation similar to the wavelet transform, yet dealing with pixels' ranks rather than with their gray-scale values. Therefore, the new FPR approach proposed herein defines a set of texture features which are calculated directly from the ranklet images corresponding to the regions of interest surviving our previous CAD system, hence, ranklet texture features; then, a support vector machine (SVM) classifier is used for discrimination. As a result of this approach, texture-based information is used to discriminate FP marks surviving our previous CAD system; at the same time, invariance to linear/nonlinear monotonic gray-scale transformations of the new CAD system is guaranteed, as ranklet texture features are calculated from ranklet images that have this property themselves by construction. To emphasize the gray scale invariance of both the previous and new CAD systems, training and testing are carried out without any in-between parameters' adjustment on mammograms having different gray-scale dynamics; in particular, training is carried out on analog digitized mammograms taken from a publicly available digital database, whereas testing is performed on full-field digital mammograms taken from an in house database. Free-response receiver operating characteristic (FROC) curve analysis of the two CAD systems demonstrates that the new approach achieves a higher reduction of FP marks when compared to the previous one. Specifically, at 60%, 65%, and 70% per-mammogram sensitivity, the new CAD system achieves 0.50, 0.68, and 0.92 FP marks per mammogram, whereas at 70%, 75%, and 80% per-case sensitivity it achieves 0.37, 0.48, and 0.71 FP marks per mammogram, respectively. Conversely, at the same sensitivities, the previous CAD system reached 0.71, 0.87, and 1.15 FP marks per mammogram, and 0.57, 0.73, and 0.92 FPs per mammogram. Also, statistical significance of the difference between the two per-mammogram and per-case FROC curves is demonstrated by the p-value < 0.001 returned by jackknife FROC analysis performed on the two CAD systems. PMID- 19291971 TI - The DOSIMAP, a high spatial resolution tissue equivalent 2D dosimeter for LINAC QA and IMRT verification. AB - The continual need for more accurate and effective techniques in radiation therapy makes it necessary to devise new control means combining high spatial resolution as well as high dose accuracy. Intensity modulated radio therapy (IMRT) allows highly conformed fields with high spatial gradient and therefore requires a precise monitoring of all the multileaf positions. In response to this need, the authors have developed a new 2D tissue equivalent dosimeter with high spatial resolution. A plastic scintillator sheet is sandwiched between two polystyrene blocks and the emitted light is captured by a high resolution camera. A newly developed procedure described herein allows efficient discrimination of the scintillation from the parasitic Cerenkov radiation. This processing is applied on the cumulated image from a sequence of images taken during an irradiation field at a rate of 10 images/s. It provides a high resolution mapping of the cumulated dose in quasireal time. The dosimeter is tissue equivalent (ICRU 44) and works both for electrons and photons without complex parameter adjustment since phantom and detector materials are identical. Instrument calibration is simple and independent of the irradiation conditions (energy, fluence, quality, ...). In this article, the authors present the principle of the dosimeter and its calibration procedure. They compare the results obtained for photons and electron beams with ionization chamber measurements in polystyrene. Technical specifications such as accuracy and repeatability are precisely evaluated and discussed. Finally, they present different IMRT field measurements and compare DOSIMAP measurements to TPS simulations and dosimetric film profiles. The results confirm the excellent spatial resolution of the instrument and its capacity to inspect the leaf positions for each segment of a given field. PMID- 19291972 TI - Deformable registration of abdominal kilovoltage treatment planning CT and tomotherapy daily megavoltage CT for treatment adaptation. AB - In adaptive radiation therapy the treatment planning kilovoltage CT (kVCT) images need to be registered with daily CT images. Daily megavoltage CT (MVCT) images are generally noisier than the kVCT images. In addition, in the abdomen, low image contrast, differences in bladder filling, differences in bowel, and rectum filling degrade image usefulness and make deformable image registration very difficult. The authors have developed a procedure to overcome these difficulties for better deformable registration between the abdominal kVCT and MVCT images. The procedure includes multiple image preprocessing steps and a two deformable registration steps. The image preprocessing steps include MVCT noise reduction, bowel gas pockets detection and painting, contrast enhancement, and intensity manipulation for critical organs. The first registration step is carried out in the local region of the critical organs (bladder, prostate, and rectum). It requires structure contours of these critical organs on both kVCT and MVCT to obtain good registration accuracy on these critical organs. The second registration step uses the first step results and registers the entire image with less intensive computational requirement. The two-step approach improves the overall computation speed and works together with these image preprocessing steps to achieve better registration accuracy than a regular single step registration. The authors evaluated the procedure on multiple image datasets from prostate cancer patients and gynecological cancer patients. Compared to rigid alignment, the proposed method improves volume matching by over 60% for the critical organs and reduces the prostate landmark registration errors by 50%. PMID- 19291973 TI - Air kerma based dosimetry calibration for the Leksell Gamma Knife. AB - No accepted official protocol exists for the dosimetry of the Leksell Gamma Knife (GK) stereotactic radiosurgery device. Establishment of a dosimetry protocol has been complicated by the unique partial-hemisphere arrangement of 201 individual 60Co beams simultaneously focused on the treatment volume and by the rigid geometry of the GK unit itself. This article proposes an air kerma based dosimetry protocol using either an in-air or in-acrylic phantom measurement to determine the absorbed dose rate of fields of the 18 mm helmet of a GK unit. A small-volume air ionization chamber was used to make measurements at the physical isocenter of three GK units. The absorbed dose rate to water was determined using a modified version of the AAPM Task Group 21 protocol designed for use with 60Co based teletherapy machines. This experimentally determined absorbed dose rate was compared to the treatment planning system (TPS) absorbed dose rate. The TPS used with the GK unit is Leksell GammaPlan. The TPS absorbed dose rate at the time of treatment is the absorbed dose rate determined by the physicist at the time of machine commissioning decay corrected to the treatment date. The TPS absorbed dose rate is defined as absorbed dose rate to water at the isocenter of a water phantom with a radius of 8 cm. Measurements were performed on model B and C Gamma Knife units. The absorbed dose rate to water for the 18 mm helmet determined using air-kerma based calculations is consistently between 1.5% and 2.9% higher than the absorbed dose rate provided by the TPS. These air kerma based measurements allow GK dosimetry to be performed with an established dosimetry protocol and without complications arising from the use of and possible variations in solid phantom material. Measurements were also made with the same ionization chamber in a spherical acrylic phantom for comparison. This methodology will allow further development of calibration methods appropriate for the smaller fields of GK units to be compared to a well established standard. PMID- 19291974 TI - Multiscale deformable registration for dual-energy x-ray imaging. AB - Dual-energy (DE) imaging of the chest improves the conspicuity of subtle lung nodules through the removal of overlying anatomical noise. Recent work has shown double-shot DE imaging (i.e., successive acquisition of low- and high-energy projections) to provide detective quantum efficiency, spectral separation (and therefore contrast), and radiation dose superior to single-shot DE imaging configurations (e.g., with a CR cassette). However, the temporal separation between high-energy (HE) and low-energy (LE) image acquisition can result in motion artifacts in the DE images, reducing image quality and diminishing diagnostic performance. This has motivated the development of a deformable registration technique that aligns the HE image onto the LE image before DE decomposition. The algorithm reported here operates in multiple passes at progressively smaller scales and increasing resolution. The first pass addresses large-scale motion by means of mutual information optimization, while successive passes (2-4) correct misregistration at finer scales by means of normalized cross correlation. Evaluation of registration performance in 129 patients imaged using an experimental DE imaging prototype demonstrated a statistically significant improvement in image alignment. Specific to the cardiac region, the registration algorithm was found to outperform a simple cardiac-gating system designed to trigger both HE and LE exposures during diastole. Modulation transfer function (MTF) analysis reveals additional advantages in DE image quality in terms of noise reduction and edge enhancement. This algorithm could offer an important tool in enhancing DE image quality and potentially improving diagnostic performance. PMID- 19291975 TI - Maximum proton kinetic energy and patient-generated neutron fluence considerations in proton beam arc delivery radiation therapy. AB - Several compact proton accelerator systems for use in proton therapy have recently been proposed. Of paramount importance to the development of such an accelerator system is the maximum kinetic energy of protons, immediately prior to entry into the patient, that must be reached by the treatment system. The commonly used value for the maximum kinetic energy required for a medical proton accelerator is 250 MeV, but it has not been demonstrated that this energy is indeed necessary to treat all or most patients eligible for proton therapy. This article quantifies the maximum kinetic energy of protons, immediately prior to entry into the patient, necessary to treat a given percentage of patients with rotational proton therapy, and examines the impact of this energy threshold on the cost and feasibility of a compact, gantry-mounted proton accelerator treatment system. One hundred randomized treatment plans from patients treated with IMRT were analyzed. The maximum radiological pathlength from the surface of the patient to the distal edge of the treatment volume was obtained for 180 degrees continuous arc proton therapy and for 180 degrees split arc proton therapy (two 90 degrees arcs) using CT# profiles from the Pinnacle (Philips Medical Systems, Madison, WI) treatment planning system. In each case, the maximum kinetic energy of protons, immediately prior to entry into the patient, that would be necessary to treat the patient was calculated using proton range tables for various media. In addition, Monte Carlo simulations were performed to quantify neutron production in a water phantom representing a patient as a function of the maximum proton kinetic energy achievable by a proton treatment system. Protons with a kinetic energy of 240 MeV, immediately prior to entry into the patient, were needed to treat 100% of patients in this study. However, it was shown that 90% of patients could be treated at 198 MeV, and 95% of patients could be treated at 207 MeV. Decreasing the proton kinetic energy from 250 to 200 MeV decreases the total neutron energy fluence produced by stopping a monoenergetic pencil beam in a water phantom by a factor of 2.3. It is possible to significantly lower the requirements on the maximum kinetic energy of a compact proton accelerator if the ability to treat a small percentage of patients with rotational therapy is sacrificed. This decrease in maximum kinetic energy, along with the corresponding decrease in neutron production, could lower the cost and ease the engineering constraints on a compact proton accelerator treatment facility. PMID- 19291976 TI - Nonrigid registration of three-dimensional ultrasound and magnetic resonance images of the carotid arteries. AB - Atherosclerosis at the carotid bifurcation can result in cerebral emboli, which in turn can block the blood supply to the brain causing ischemic strokes. Noninvasive imaging tools that better characterize arterial wall, and atherosclerotic plaque structure and composition may help to determine the factors which lead to the development of unstable lesions, and identify patients at risk of plaque disruption and stroke. Carotid magnetic resonance (MR) imaging allows for the characterization of carotid vessel wall and plaque composition, the characterization of normal and pathological arterial wall, the quantification of plaque size, and the detection of plaque integrity. On the other hand, various ultrasound (US) measurements have also been used to quantify atherosclerosis, carotid stenosis, intima-media thickness, total plaque volume, total plaque area, and vessel wall volume. Combining the complementary information provided by 3D MR and US carotid images may lead to a better understanding of the underlying compositional and textural factors that define plaque and wall vulnerability, which may lead to better and more effective stroke prevention strategies and patient management. Combining these images requires nonrigid registration to correct the nonlinear misalignments caused by relative twisting and bending in the neck due to different head positions during the two image acquisition sessions. The high degree of freedom and large number of parameters associated with existing nonrigid image registration methods causes several problems including unnatural plaque morphology alteration, high computational complexity, and low reliability. Thus, a "twisting and bending" model was used with only six parameters to model the normal movement of the neck for nonrigid registration. The registration technique was evaluated using 3D US and MR carotid images at two field strengths, 1.5 and 3.0 T, of the same subject acquired on the same day. The mean registration error between the segmented carotid artery wall boundaries in the target US image and the registered MR images was calculated using a distance based error metric after applying a "twisting and bending" model based nonrigid registration algorithm. An average registration error of 1.4 +/- 0.3 mm was obtained for 1.5 T MR and 1.5 +/- 0.4 mm for 3.0 T MR, when registered with 3D US images using the nonrigid registration technique presented in this paper. Visual inspection of segmented vessel surfaces also showed a substantial improvement of alignment with this nonrigid registration technique compared to rigid registration. PMID- 19291977 TI - Accuracy in the localization of thoracic and abdominal tumors using respiratory displacement, velocity, and phase. AB - Current four dimensional (4D) computed tomography (CT) reconstruction techniques are retrospectively created based on either the phase or displacement of the respiratory signal. Both techniques have known limitations which cause clinically significant motion artifacts in 4D CT images. These artifacts, which appear as undefined or irregular boundaries in the 4D CT images, cause systematic errors in patient contouring and dose calculations. The aim of this work was to evaluate the reproducibility of tumor position as a function of displacement, phase, and velocity of the respiratory signal, respectively, in order to determine the optimum parameter or combination of parameters to use in order to minimize artifacts in 4D CT images or to accurately deliver radiation to relevant structures during treatment. METHOD AND MATERIALS: Estimated tumor centroid position and respiratory signal data were acquired with the Cyberknife Synchrony system for 26 thoracic radiotherapy patients (52 fractions). A reference respiratory cycle was calculated for each patient. Displacement, phase, and velocity of ten data points were calculated from this reference respiratory cycle, where each point represents an image bin. The corresponding tumor position was then sorted into these image bins if the phase, displacement, simultaneous displacement and phase, or simultaneous displacement and velocity of the respiratory signal were within tolerances of 0.5 mm for displacement and 0.5 mm/s for velocity, respectively, from the corresponding data of the reference cycle for each image bin. RESULTS: The mean of the standard deviations of tumor positions over all bins and all fractions for the superior-inferior direction were 2.13 +/- 1.01 mm for phase sorting, 1.20 +/- 0.76 mm for displacement sorting, 1.20 +/- 0.71 mm for simultaneous displacement and phase sorting, and 1.10 +/- 0.71 mm for simultaneous displacement and velocity sorting, with maximum deviations of 43.0, 16.1, 15.5, and 14.1 mm for each scenario, respectively. The same trend was observed for the anterior-posterior and left-right directions. A linear dependence was observed between the mean of the standard deviations of tumor positions over all fractions as a function of the velocity of the respiratory signal at each bin for all the sorting scenarios. A substantially larger gradient for the phase sorting scenario, compared to the other scenarios, suggests that tumor localization will become increasingly less accurate as the velocity of the tumor increases during a breathing cycle, e.g., if the amplitude of motion increases while the period of the respiratory cycle stays constant or during mid inhale or exhale phases of the respiratory cycle. CONCLUSION: This study illustrates that position of a tumor can be determined more accurately if displacement and velocity are used simultaneously as sorting parameters for 4D CT images or during treatment. A real-time displacement and velocity based 4D CT image sorting method may therefore produce fewer and smaller artifacts in 4D CT images than current retrospective sorting methods. PMID- 19291978 TI - Direct two-dimensional reconstruction algorithm for an inverse-geometry CT system. AB - An inverse-geometry computed tomography (IGCT) system uses a large source array opposite a smaller detector array. A previously described IGCT reconstruction algorithm uses gridding, but this gridding step produces blurring in the reconstructed image. In this article, the authors describe a two-dimensional (2D) IGCT reconstruction algorithm without gridding. In the transverse direction, the raw data of the IGCT system can be viewed as being composed of many fan beams. Because the spacing between source spots is larger than the spot width, each fan beam has undersampled projection data, but the missing samples are effectively provided by other undersampled fan beam views. In the proposed method, a direct fan beam reconstruction algorithm is used to process each undersampled fan beam. Initial images with this method showed ring artifacts caused by nonuniform sampling in the radial direction as compared to an ideal fan beam. A new method for correcting this effect was developed. With this correction, high quality images were obtained. The noise performance of the proposed 2D IGCT reconstruction algorithm was investigated, and it was comparable to that of the fan beam system. A MTF study showed that the proposed method achieves better resolution than the gridding method. PMID- 19291979 TI - Optical observation of energy loss distribution and practical range of positrons from a 18F water solution in a water-equivalent phantom. AB - The energy loss distribution of beta+ particles is closely related to their maximum penetration depth distribution and annihilation point distribution. The latter is of practical importance for positron emission tomography. Experimental data related to the energy loss distribution are important for comprehensive validation of physics and simulation models of beta+ interactions. In this paper the authors present a new experimental approach that allows them to visually observe the beta+ energy loss distribution of a solution of nuclear medicine radioisotopes in a plastic scintillator using an optical camera. The authors also report a set of the first experimental results. A water solution of 18F was localized in a small hole in a plastic scintillator (BC430). Optical imaging of the scintillator yielded visual images of the energy loss distribution with a submillimeter resolution. The radial dependence in the energy distribution was quantitatively measured by analysis of the images, and exponential fitting parameters were obtained. The authors observed that the results of Monte Carlo simulation with EGS5 (version 1.0.2) and GEANT4 (version 4.9.01.p01) were consistent with those obtained experimentally. The results of the Monte Carlo simulation indicated that for a linear scale, the energy loss distribution in the scintillator was approximately the same as that in water, and the relative shape of the energy loss distribution was close to those of the maximum penetration depth distribution and annihilation point distribution. This paper also presents discussions about the further possibilities of this optical imaging approach. Thus, optical observation of the beta+ energy loss distribution in a scintillator is a promising technique for visual and quantitative experimental studies of beta+ emission from a solution of radioisotopes that are used in nuclear medicine. PMID- 19291980 TI - Anniversary paper: evolution of ultrasound physics and the role of medical physicists and the AAPM and its journal in that evolution. AB - Ultrasound has been the greatest imaging modality worldwide for many years by equipment purchase value and by number of machines and examinations. It is becoming increasingly the front end imaging modality; serving often as an extension of the physician's fingers. We believe that at the other extreme, high end systems will continue to compete with all other imaging modalities in imaging departments to be the method of choice for various applications, particularly where safety and cost are paramount. Therapeutic ultrasound, in addition to the physiotherapy practiced for many decades, is just coming into its own as a major tool in the long progression to less invasive interventional treatment. The physics of medical ultrasound has evolved over many fronts throughout its history. For this reason, a topical review, rather than a primarily chronological one is presented. A brief review of medical ultrasound imaging and therapy is presented, with an emphasis on the contributions of medical physicists, the American Association of Physicists in Medicine (AAPM) and its publications, particularly its journal Medical Physics. The AAPM and Medical Physics have contributed substantially to training of physicists and engineers, medical practitioners, technologists, and the public. PMID- 19291981 TI - Optimizing the dynamic range extension of a radiochromic film dosimetry system. AB - The authors present a radiochromic film dosimetry protocol for a multicolor channel radiochromic film dosimetry system consisting of the external beam therapy (EBT) model GAFCHROMIC film and the Epson Expression 1680 flat-bed document scanner. Instead of extracting only the red color channel, the authors are using all three color channels in the absorption spectrum of the EBT film to extend the dynamic dose range of the radiochromic film dosimetry system. By optimizing the dose range for each color channel, they obtained a system that has both precision and accuracy below 1.5%, and the optimized ranges are 0-4 Gy for the red channel, 4-50 Gy for the green channel, and above 50 Gy for the blue channel. PMID- 19291982 TI - Threshold adjusted calcium scoring using CT is less susceptible to cardiac motion and more accurate. AB - The purpose of this paper is to investigate calcium scoring on computed tomography (CT) using an adjusted threshold depending on the maximum Hounsfield value within the calcification (HU(peak)). The volume of 19 calcifications was retrospectively determined on 64-slice multidetector CT and dual source CT (DSCT) at different thresholds and the threshold associated with the physical volume was determined. In addition, approximately 10 000 computer simulations were done simulating the same process for calcifications with mixed density. Using these data a relation between the HU(peak) and the threshold could be established. Hereafter, this relation was assessed by scanning six calcifications in a phantom at 40-110 beats per minute using DSCT. The influence of motion was determined and the measured calcium scores were compared to the physical volumes and mass. A positive linear correlation was found between the scoring threshold and the HU(peak) of the calcifications both for the phantom measurements as for the computer simulations. Using this relation the individual threshold for each calcification could be calculated. Calcium scores of the moving calcifications determined with an adjusted threshold were approximately 30% less susceptible to cardiac motion compared to standard calcium scoring. Furthermore, these scores approximated the physical volume and mass at least 10% better than the standard calcium scores. The threshold in calcium scoring should be adjusted for each individual calcification based on the HU(peak) of the calcification. Calcium scoring using an adjusted threshold is less susceptible to cardiac motion and more accurate compared to the physical values. PMID- 19291983 TI - Microbeam radiation therapy: a Monte Carlo study of the influence of the source, multislit collimator, and beam divergence on microbeams. AB - Microbeam radiation therapy (MRT) is a new oncology method currently under development for the treatment of inoperable pediatric brain tumors. Monte Carlo simulation, or the computational study of radiation transport in matter, is often used in radiotherapy to theoretically estimate the dose required for treatment. However, its potential use in MRT dose planning systems is currently hindered by the significant discrepancies that have been observed between measured and theoretical dose and the PVDR (peak to valley dose ratio). The need to resolve these discrepancies is driven by the desirability of making MRT available to humans in the next few years. This article aims to resolve some of the discrepancies by examining the simplifications adopted in previous MRT Monte Carlo studies, such as the common practice of commencing microbeam transport on the surface of the target which neglects the influence of the distributed synchrotron source, multislit collimator, and the beam divergence between them. This article uses PENELOPE Monte Carlo simulation to investigate the influence of these beamline components upstream of the target on the lateral dose profiles and PVDRs of an array of 25 microbeams. It also compares the dose profiles and PVDRs of a microbeam array produced from a single simulation (full array) to those produced from the superposition of a single microbeam profile (sup array). The effect of modeling the distributed source and the beam divergence was an increase in the absorbed dose in the penumbral and valley regions of the microbeam profiles. Inclusion of the multislit collimator resulted in differences of up to 5 microm in the FWHM of microbeam profiles across the array, which led to minor variations in the corresponding PVDR yields. PMID- 19291984 TI - Atlas-based functional radiosurgery: early results. AB - Functional disorders of the brain, such as dystonia and neuropathic pain, may respond poorly to medical therapy. Deep brain stimulation (DBS) of the globus pallidus pars interna (GPi) and the centromedian nucleus of the thalamus (CMN) may alleviate dystonia and neuropathic pain, respectively. A noninvasive alternative to DBS is radiosurgical ablation [internal pallidotomy (IP) and medial thalamotomy (MT)]. The main technical limitation of radiosurgery is that targets are selected only on the basis of MRI anatomy, without electrophysiological confirmation. This means that, to be feasible, image-based targeting must be highly accurate and reproducible. Here, we report on the feasibility of an atlas-based approach to targeting for functional radiosurgery. In this method, masks of the GPi, CMN, and medio-dorsal nucleus were nonrigidly registered to patients' T1-weighted MRI (T1w-MRI) and superimposed on patients' T2-weighted MRI (T2w-MRI). Radiosurgical targets were identified on the T2w-MRI registered to the planning CT by an expert functional neurosurgeon. To assess its feasibility, two patients were treated with the CyberKnife using this method of targeting; a patient with dystonia received an IP (120 Gy prescribed to the 65% isodose) and a patient with neuropathic pain received a MT (120 Gy to the 77% isodose). Six months after treatment, T2w-MRIs and contrast-enhanced T1w-MRIs showed edematous regions around the lesions; target placements were reevaluated by DW-MRIs. At 12 months post-treatment steroids for radiation-induced edema and medications for dystonia and neuropathic pain were suppressed. Both patients experienced significant relief from pain and dystonia-related problems. Fifteen months after treatment edema had disappeared. Thus, this work shows promising feasibility of atlas-based functional radiosurgery to improve patient condition. Further investigations are indicated for optimizing treatment dose. PMID- 19291985 TI - Compression and smart coding of offset and gain maps for intraoral digital x-ray sensors. AB - The response of indirect x-ray digital imaging sensors is often not homogenous on the entire surface area. In this case, calibration is needed to build offset and gain maps, which are used to correct the sensor output. The sensors of new generation are equipped with an on-board memory, which serves to store these maps. However, because of its limited dimension, the maps have to be compressed before saving them. This step is critical because of the extremely high compression rate required. The authors propose here a novel method to achieve such a high compression rate, without degrading the quality of the sensor output. It is based on quad tree decomposition, which performs an adaptive sampling of the offset and gain maps, matched with a RBF-based interpolation strategy. The method was tested on a typical intraoral radiographic sensor and compared with traditional compression techniques. Qualitative and quantitative results show that the method achieves a higher compression rate and produces images of superior quality. The method can be adopted also in different fields where a high compression rate is required. PMID- 19291986 TI - A modified gradient correlation filter for image segmentation: application to airway and bowel. AB - The segmentation of structures of interest from medical images may incorrectly include adjacent structures in the segmented image (i.e., false positives). This study introduces a family of gradient correlation filters that reduce false positives in the segmented image by comparing the segmented region gradients with a user-defined model. A gradient correlation filter was applied to a database of clinical computed tomography scans for the task of differentiating airway from lung regions and bowel from lung regions. The results were evaluated using receiver-operating characteristic analysis and demonstrated excellent results for both the airway/lung and bowel/lung classification tasks. PMID- 19291987 TI - Monte Carlo radiation dose simulations and dosimetric comparison of the model 6711 and 9011 125I brachytherapy sources. AB - Smaller diameter brachytherapy seeds for permanent interstitial implantation allow for use of smaller diameter implant needles. The use of smaller diameter needles may provide a lower incidence of healthy-tissue complications. This study determines the brachytherapy dosimetry parameters for the smaller diameter source (model 9011) and comments on the dosimetric comparison between this new source and the conventional brachytherapy seed (model 6711). PMID- 19291988 TI - Statistical characteristics of streak artifacts on CT images: relationship between streak artifacts and mA s values. AB - The purpose of this study is to investigate how streak artifacts on computed tomography (CT) images vary with reduction in radiation doses by assessing the quantitative relationship between the streak artifacts and milliampere-time product (mA s) values. A commercially available chest phantom was used to measure the streak artifacts on the CT images obtained using a 4- and 16-multidetector row helical CT scanners with various mA s values at a constant tube voltage of 120 kVp. The cardiac slice image was employed as a target image for evaluating the streak artifacts on the CT image. Eighty parallel line segments with a length of 20 pixels were placed perpendicular to numerous streak artifacts on the cardiac slice image, and the largest difference between adjacent CT values in each of the 80 CT-value profiles of these line segments was employed as a feature variable of streak artifacts; these feature variables have been analyzed by the extreme value theory. The largest difference between adjacent CT values in each CT-value profile can be statistically modeled by a Gumbel distribution. Further, the maximum level of streak artifacts on CT images that will be tolerated for clinical use and low-dose CT screening examination was expected to be estimated using the location parameter in the Gumbel distribution. PMID- 19291989 TI - A local shift-variant Fourier model and experimental validation of circular cone beam computed tomography artifacts. AB - Large field of view cone-beam computed tomography (CBCT) is being achieved using circular source and detector trajectories. These circular trajectories are known to collect insufficient data for accurate image reconstruction. Although various descriptions of the missing information exist, the manifestation of this lack of data in reconstructed images is generally nonintuitive. One model predicts that the missing information corresponds to a shift-variant cone of missing frequency components. This description implies that artifacts depend on the imaging geometry, as well as the frequency content of the imaged object. In particular, objects with a large proportion of energy distributed over frequency bands that coincide with the missing cone will be most compromised. These predictions were experimentally verified by imaging small, localized objects (acrylic spheres, stacked disks) at varying positions in the object space and observing the frequency spectrums of the reconstructions. Measurements of the internal angle of the missing cone agreed well with theory, indicating a right circular cone for points on the rotation axis, and an oblique, circular cone elsewhere. In the former case, the largest internal angle with respect to the vertical axis corresponds to the (half) cone angle of the CBCT system (typically approximately 5 degrees - 7.5 degrees in IGRT). Object recovery was also found to be strongly dependent on the distribution of the object's frequency spectrum relative to the missing cone, as expected. The observed artifacts were also reproducible via removal of local frequency components, further supporting the theoretical model. Larger objects with differing internal structures (cellular polyurethane, solid acrylic) were also imaged and interpreted with respect to the previous results. Finally, small animal data obtained using a clinical CBCT scanner were observed for evidence of the missing cone. This study provides insight into the influence of incomplete data collection on the appearance of objects imaged in large field of view CBCT. PMID- 19291990 TI - In vitro in-stent restenoses evaluated by 3D ultrasound. AB - The purpose of this study was to quantify in-stent restenoses with 3D B mode and power Doppler ultrasound (U.S.) imaging. In-stent restenoses were mimicked with vascular phantoms in which a nonferromagnetic prototype stent (Boston Scientific) and a ferromagnetic clinical stainless steel stent (Palmaz P295) were embedded. Each phantom had an 80% in-stent stenosis and a 75% stenosis located outside the stent. These phantoms were compared to a reference phantom reproducing both stenoses without stent. Data sets of 2D cross-sectional U.S. images were acquired in freehand scanning using a magnetic sensor attached to the U.S. probe and in mechanical linear scanning with the probe attached to a step motor device. Each 2D image was automatically segmented before 3D reconstruction of the vessel. Results indicate that the reference phantom (without stent) was accurately assessed with errors below 1.8% for the 75% stenosis and 3.2% for the 80% stenosis in both B mode and power Doppler for the two scanning methods. The 80% in-stent stenoses in Boston Scientific and Palmaz stents were, respectively, evaluated at 73.8 (+/-3.2)% and 75.8 (+/- 3)% in B mode and at 82 (+/- 2.5)% and 86.2 (+/- 6.4)% in power Doppler when freehand scans were used. For comparison, when linear scans were selected, in-stent stenoses in the Boston Scientific or Palmaz stent were, respectively, evaluated at 77.4 (+/- 2.0)% and 73.8 (+/- 2.5)% in B mode and at 87.0 (+/- 1.3)% and 85.6 (+/- 5.8)% in power Doppler. To conclude, 3D freehand U.S. is a valuable method to quantify in-stent restenoses, particularly in B mode. It is thus hoped that, in the clinical setting, noninvasive 3D U.S. may provide sufficient precision to grade in-stent restenoses. PMID- 19291991 TI - Determination of correction factors for a 2D diode array device in MV photon beams. AB - The purpose of this study is to evaluate the dosimetric characteristics of a 2D diode array device (MAPCHECK) and provide correction factors for clinically relevant conditions. Measurements were performed for various collimator settings (5 x 5-40 x 40 cm2), SSDs (80-110 cm), and depths (5-20 cm) for 6 and 15 MV photon beams. The correction factor was determined as the ratio of dose measured by an ionization chamber in a large full scatter water phantom to the dose measured by the MAPCHECK with 30 x 30 cm2 solid water phantom. The total correction factor was separated into three components. The phantom scatter factor was evaluated by taking the ratio of dose measured by MAPCHECK with backscatter material and 30 x 30 cm2 phantom to that by the ionization chamber in an infinite large water phantom renormalized to the reference field size (10 x 10 cm2). This was also compared to the theoretical calculation using ratios of phantom scatter factor between an infinitely large water phantom and 30 x 30 cm2 solid water phantoms. The backscatter effect was determined by taking the ratio of the MAPCHECK measurements with and without 10 cm thick solid water backscatter material also normalized to the reference field size at the depth of measurement. The extra correction factor is defined as the ratio of dose measured by the MAPCHECK (without backscatter) to that measured by the ionization chamber in a large water phantom at depth d for the reference field size (10 x 10 cm2) normalized to the reference depth (10 cm) and reference field size. The total correction factors for 6 and 15 MV photon beams were up to 16% and 11.5%, respectively, due to the difference in phantom scatter in MAPCHECK measurements when the field sizes are larger than 30 cm. The result was consistent with theoretical analysis with phantom scatter alone. The backscatter correction factor varied by up to 2% and increased with increasing field size. The extra correction factor decreases with depths and is < 0.4% for all depths. The authors have verified that correction factors for MAPCHECK are a function of the scaled field size and depth only and are independent of the SSD. Results were verified for off-axis points measured by MAPCHECK. PMID- 19291992 TI - Considerations and limitations of fast Monte Carlo electron transport in radiation therapy based on precalculated data. AB - The purpose of this work is to revisit the impediments and characteristics of fast Monte Carlo techniques for applications in radiation therapy treatment planning using new methods of utilizing pregenerated electron tracks. The limitations of various techniques for the improvement of speed and accuracy of electron transport have been evaluated. A method is proposed that takes advantage of large available memory in current computer hardware for extensive generation of precalculated data. Primary tracks of electrons are generated in the middle of homogeneous materials (water, air, bone, lung) and with energies between 0.2 and 18 MeV using the EGSnrc code. Secondary electrons are not transported, but their position, energy, charge, and direction are saved and used as a primary particle. Based on medium type and incident electron energy, a track is selected from the precalculated set. The performance of the method is tested in various homogeneous and heterogeneous configurations and the results were generally within 2% compared to EGSnrc but with a 40-60 times speed improvement. In a second stage the authors studied the obstacles for further increased speed-ups in voxel geometries by including ray-tracing and particle fluence information in the pregenerated track information. The latter method leads to speed increases of about a factor of 500 over EGSnrc for voxel-based geometries. In both approaches, no physical calculation is carried out during the runtime phase after the pregenerated data has been stored even in the presence of heterogeneities. The precalculated data are generated for each particular material and this improves the performance of the precalculated Monte Carlo code both in terms of accuracy and speed. Precalculated Monte Carlo codes are accurate, fast, and physics independent and therefore applicable to different radiation types including heavy charged particles. PMID- 19291993 TI - Comparison of patient size-based methods for estimating quantum noise in CT images of the lung. AB - The authors explored four methods for estimating quantum noise in CT images of the lung, each based on a different definition of patient size (water-equivalent diameter) and the relationship between noise and diameter determined in water phantoms. The accuracies of the four methods were evaluated using an image subtraction method as a gold standard. Noise estimates based on patient sizes derived from chest area, water-equivalent area, non-lung area, and water equivalent path length had maximum errors of 229%, 93%, 34%, and 57%, respectively. Considering the magnitude of noise variation across the lung volume (approximately 30%), noise estimate based on non-lung area was reasonably accurate. PMID- 19291994 TI - An energy fluence-convolution model for amorphous silicon EPID dose prediction. AB - In this work, an amorphous silicon electronic portal imaging device (a-Si EPID) dose prediction model based on the energy fluence model of the Pinnacle treatment planning system Version 7 (Philips Medical Systems, Madison, WI) is developed. An energy fluence matrix at very high resolution (< 1 mm) is used to incorporate multileaf collimator (MLC) leaf effects in the predicted EPID images. The primary dose deposited in the EPID is calculated from the energy fluence using experimentally derived radially dependent EPID interaction coefficients. Separate coefficients are used for the open beam energy fluence component and the component of the energy fluence transmitted through closed MLC leaves to each EPID pixel. A spatially invariant EPID dose deposition kernel that describes both radiative dose deposition, central axis EPID backscatter, and optical glare is convolved with the primary dose. The kernel is further optimized to give accurate EPID penumbra prediction and EPID scatter factor with changing MLC field size. An EPID calibration method was developed to reduce the effect of nonuniform backscatter from the support arm (E-arm) in a calibrated EPID image. This method removes the backscatter component from the pixel sensitivity (flood field) correction matrix retaining only field-specific backscatter in the images. The model was compared to EPID images for jaw and MLC defined open fields and eight head and neck intensity modulated radiotherapy (IMRT) fields. For the head and neck IMRT fields with 2%, 2 mm criteria 97.6 +/- 0.6% (mean +/- 1 standard deviation) of points passed with a gamma index less than 1, and for 3%, 3 mm 99.4 +/- 0.4% of points were within the criteria. For these fields, the 2%, 2 mm pass score reduced to 96.0 +/- 1.5% when backscatter was present in the pixel sensitivity correction matrix. The model incorporates the effect of MLC leaf transmission, EPID response to open and MLC leakage dose components, and accurately predicts EPID images of IMRT fields. Removing the backscatter component of the pixel sensitivity matrix correction reduces the effect of nonuniform E-arm backscatter. PMID- 19291995 TI - Vision 20/20: proton therapy. AB - The first patients were treated with proton beams in 1955 at the Lawrence Berkeley Laboratory in California. In 1970, proton beams began to be used in research facilities to treat cancer patients using fractionated treatment regimens. It was not until 1990 that proton treatments were carried out in hospital-based facilities using technology and techniques that were comparable to those for modern photon therapy. Clinical data strongly support the conclusion that proton therapy is superior to conventional radiation therapy in a number of disease sites. Treatment planning studies have shown that proton dose distributions are superior to those for photons in a wide range of disease sites indicating that additional clinical gains can be achieved if these treatment plans can be reliably delivered to patients. Optimum proton dose distributions can be achieved with intensity modulated protons (IMPT), but very few patients have received this advanced form of treatment. It is anticipated widespread implementation of IMPT would provide additional improvements in clinical outcomes. Advances in the last decade have led to an increased interest in proton therapy. Currently, proton therapy is undergoing transitions that will move it into the mainstream of cancer treatment. For example, proton therapy is now reimbursed, there has been rapid development in proton therapy technology, and many new options are available for equipment, facility configuration, and financing. During the next decade, new developments will increase the efficiency and accuracy of proton therapy and enhance our ability to verify treatment planning calculations and perform quality assurance for proton therapy delivery. With the implementation of new multi-institution clinical studies and the routine availability of IMPT, it may be possible, within the next decade, to quantify the clinical gains obtained from optimized proton therapy. During this same period several new proton therapy facilities will be built and the cost of proton therapy is expected to decrease, making proton therapy routinely available to a larger population of cancer patients. PMID- 19291996 TI - Measurement of compressed breast thickness by optical stereoscopic photogrammetry. AB - The determination of volumetric breast density (VBD) from mammograms requires accurate knowledge of the thickness of the compressed breast. In attempting to accurately determine VBD from images obtained on conventional mammography systems, the authors found that the thickness reported by a number of mammography systems in the field varied by as much as 15 mm when compressing the same breast or phantom. In order to evaluate the behavior of mammographic compression systems and to be able to predict the thickness at different locations in the breast on patients, they have developed a method for measuring the local thickness of the breast at all points of contact with the compression paddle using optical stereoscopic photogrammetry. On both flat (solid) and compressible phantoms, the measurements were accurate to better than 1 mm with a precision of 0.2 mm. In a pilot study, this method was used to measure thickness on 108 volunteers who were undergoing mammography examination. This measurement tool will allow us to characterize paddle surface deformations, deflections and calibration offsets for mammographic units. PMID- 19291997 TI - Accurate estimation of compressed breast thickness in mammography. AB - The determination of volumetric breast density (VBD) from mammograms requires an accurate knowledge of the thickness of the compressed breast. Previously, the authors described a technique for measuring local thicknesses using optical stereoscopic photogrammetry [A. H. Tyson, G. E. Mawdsley, and M. J. Yaffe, "Measurement of compressed breast thickness by optical stereoscopic photogrammetry," Med. Phys. 36(2), 569-576 (2009)]. Here, the authors describe the use of this tool to guide the development of a simpler, more practical field technique for the estimation of breast thickness and test its accuracy. Phantoms were constructed having similar shapes and compression characteristics to breasts of different sizes. These phantoms were compressed at different forces on several types of mammography units and their thickness under compression was measured using optical stereoscopic photogrammetry at many points of contact with the compression plate. A prediction equation was developed that uses the readout of compressed thickness and compression force provided by the mammography system to estimate local breast thickness. Using this approach, systems can be calibrated to an accuracy of better than 5 mm in thickness using a simple test object compared to an error of up to 15 mm associated with using only the thickness readout of the mammography machine. On the systems tested, the estimated value of VBD obtained using this method is significantly reduced from that determined using the constant thickness reported by the mammography machine. PMID- 19291998 TI - Measurement of the x-ray tube anodes' surface profile and its effects on the x ray spectra. AB - An experimental study--involving measurements with an optical microscope, a profilometer, and a scanning electron microscope--for determination of the surface profile of x-ray tube anodes is presented. The islands on the "mud flatting" surface are separated by approximately 8 microm deep cracks. The surface roughness on the island is typically below 1 microm, and the area ratio of cracks to the total surface is higher on the more extensively used regions (anode aging). A simple model was proposed to calculate the spectrum modification introduced by the rough surface. Loss of x-ray intensity of 4% was predicted using the roughest surface at a small emission angle. PMID- 19291999 TI - Depth dependence of electron backscatter: an energy spectral and dosimetry study using Monte Carlo simulation. AB - This study investigated the depth dependence of electron backscatter from a layer of lead (Pb) for clinical electron beams. The change in the electron backscatter with variation in the water depth above the Pb was determined. Electron energy spectra and relative depth doses as a function of depth in water over the Pb layer were calculated using a Monte Carlo simulation and studied. Phase-space files for 4 and 9 MeV electron beams (10 x 10 cm2 applicator and cutout) based on the Varian 21 EX linear accelerator were generated using the EGSnrc-based BEAMNRC code. 3 mm of Pb, at depths of 0.5 and 1 cm in water, was irradiated with electrons. The source-to-surface distance is equal to 100 cm. Electron energy spectra and relative depth doses with and without the presence of the Pb layer at different depths in water were determined using the BEAMNRC code. For the 4 MeV electron energy spectra at a depth of 0.5 cm in water, electron backscatter was found to originate at the Pb-water interface and extend to 0.5 cm above the Pb insert. However, at a depth of 1 cm in water, electron backscatter almost disappeared at 0.5 and 1 cm above th ePb insert. This is due to the increased attenuation of the incident 4 MeV electron beam in a thicker layer of water as well as increased attenuation of the electron backscatter above the Pb. This resulted in a 23% decrease in relative dose at a measurement point of 0.5 cm depth, when the depth of the Pb insert was changed from 1 to 0.5 cm. For the electron energy spectra of the 9 MeV beams with a 0.5 cm depth of water, only a small amount of electron backscatter was observed. However, more electron backscatter was found when the water depth was increased to 1 cm. This is because the electron beam energy was decreased more due to the increase in attenuation from the increased depth of water compared to 0.5 cm. Since the electron energy spectrum and relative depth dose above the Pb layer vary with depth of water on top of the Pb, the electron backscatter depends significantly on the thickness of water, or water equivalent bolus, or critical tissue over the Pb shield in electron radiotherapy. PMID- 19292000 TI - Quantitative simultaneous 99mTc/123I cardiac SPECT using MC-JOSEM. AB - Simultaneous rest 99mTc-Sestamibi/ 123I-BMIPP cardiac SPECT imaging has the potential to replace current clinical 99mTc-Sestamibi rest/stress imaging and therefore has great potential in the case of patients with chest pain presenting to the emergency department. Separation of images of these two radionuclides is difficult, however, because their emission energies are close. The authors previously developed a fast Monte Carlo (MC)-based joint ordered-subset expectation maximization (JOSEM) iterative reconstruction algorithm (MC-JOSEM), which simultaneously compensates for scatter and cross talk as well as detector response within the reconstruction algorithm. In this work, the authors evaluated the performance of MC-JOSEM in a realistic population of 99mTc/123I studies using cardiac phantom data on a Siemens e.cam system using a standard cardiac protocol. The authors also compared the performance of MC-JOSEM for estimation tasks to that of two other methods: standard OSEM using photopeak energy windows without scatter correction (NSC-OSEM) and standard OSEM using a Compton-scatter energy window for scatter correction (SC-OSEM). For each radionuclide the authors separately acquired high-count projections of radioactivity in the myocardium wall, liver, and soft tissue background compartments of a water-filled torso phantom, and they generated synthetic projections of various dual-radionuclide activity distributions. Images of different combinations of myocardium wall/background activity concentration ratios for each radionuclide were reconstructed by NSC-OSEM, SC-OSEM, and MC-JOSEM. For activity estimation in the myocardium wall, MC-JOSEM always produced the best relative bias and relative standard deviation compared with NSC-OSEM and SC-OSEM for all the activity combinations. On average, the relative biases after 100 iterations were 8.1% for 99mTc and 3.7% for 123I with MC-JOSEM, 39.4% for 99mTc and 23.7% for 123I with NSC-OSEM, and 20.9% for 99mTc with SC-OSEM. The relative standard deviations after 30 iterations were 0.7% for 99mTc and 1.0% for 123I with MC-JOSEM, as compared to 1.1% for 99mTc and 1.2% for 123I with NSC-OSEM and 1.3% for 99mTc with SC-OSEM. Finally, the authors compared the relative standard deviation after 30 iterations with the minimum theoretical variance on activity estimation, the Cramer-Rao lower bound (CRB), and with the biased CRB. The measured precision was larger than the biased bound values by factors of 2-4, suggesting that further improvement could be made to the method. PMID- 19292001 TI - Evaluation of quantitative imaging methods for organ activity and residence time estimation using a population of phantoms having realistic variations in anatomy and uptake. AB - Estimating organ residence times is an essential part of patient-specific dosimetry for radioimmunotherapy (RIT). Quantitative imaging methods for RIT are often evaluated using a single physical or simulated phantom but are intended to be applied clinically where there is variability in patient anatomy, biodistribution, and biokinetics. To provide a more relevant evaluation, the authors have thus developed a population of phantoms with realistic variations in these factors and applied it to the evaluation of quantitative imaging methods both to find the best method and to demonstrate the effects of these variations. Using whole body scans and SPECT/CT images, organ shapes and time-activity curves of 111In ibritumomab tiuxetan were measured in dosimetrically important organs in seven patients undergoing a high dose therapy regimen. Based on these measurements, we created a 3D NURBS-based cardiac-torso (NCAT)-based phantom population. SPECT and planar data at realistic count levels were then simulated using previously validated Monte Carlo simulation tools. The projections from the population were used to evaluate the accuracy and variation in accuracy of residence time estimation methods that used a time series of SPECT and planar scans, Quantitative SPECT (QSPECT) reconstruction methods were used that compensated for attenuation, scatter, and the collimator-detector response. Planar images were processed with a conventional (CPlanar) method that used geometric mean attenuation and triple-energy window scatter compensation and a quantitative planar (QPlanar) processing method that used model-based compensation for image degrading effects. Residence times were estimated from activity estimates made at each of five time points. The authors also evaluated hybrid methods that used CPlanar or QPlanar time-activity curves rescaled to the activity estimated from a single QSPECT image. The methods were evaluated in terms of mean relative error and standard deviation of the relative error in the residence time estimates taken over the phantom population. The mean errors in the residence time estimates over all the organs were < 9.9% (pure QSPECT), < 13.2% (pure QPLanar), < 7.2% (hybrid QPlanar/QSPECT), < 19.2% (hybrid CPlanar/QSPECT), and 7%-159% (pure CPlanar). The standard deviations of the errors for all the organs over all the phantoms were < 9.9%, < 11.9%, < 10.8%, < 22.0%, and < 107.9% for the same methods, respectively. The processing methods differed both in terms of their average accuracy and the variation of the accuracy over the population of phantoms, thus demonstrating the importance of using a phantom population in evaluating quantitative imaging methods. Hybrid CPlanar/QSPECT provided improved accuracy compared to pure CPlanar and required the addition of only a single SPECT acquisition. The QPlanar or hybrid QPlanar/QSPECT methods had mean errors and standard deviations of errors that approached those of pure QSPECT while providing simplified image acquisition protocols, and thus may be more clinically practical. PMID- 19292002 TI - An investigation on the capabilities of the PENELOPE MC code in nanodosimetry. AB - The Monte Carlo (MC) method has been widely implemented in studies of radiation effects on human genetic material. Most of these works have used specific-purpose MC codes to simulate radiation transport in condensed media. PENELOPE is one of the general-purpose MC codes that has been used in many applications related to radiation dosimetry. Based on the fact that PENELOPE can carry out event-by-event coupled electron-photon transport simulations following these particles down to energies of the order of few tens of eV, we have decided to investigate the capacities of this code in the field of nanodosimetry. Single and double strand break probabilities due to the direct impact of gamma rays originated from Co60 and Cs137 isotopes and characteristic x-rays, from Al and C K-shells, have been determined by use of PENELOPE. Indirect damage has not been accounted for in this study. A human genetic material geometrical model has been developed, taking into account five organizational levels. In an article by Friedland et al. [Radiat. Environ. Biophys. 38, 39-47 (1999)], a specific-purpose MC code and a very sophisticated DNA geometrical model were used. We have chosen that work as a reference to compare our results. Single and double strand-break probabilities obtained here underestimate those reported by Friedland and co-workers by 20%-76% and 50%-60%, respectively. However, we obtain RBE values for Cs137, AlK and CK radiations in agreement with those reported in previous works [Radiat. Environ. Biophys. 38, 39-47 (1999)] and [Phys. Med. Biol. 53, 233-244 (2008)]. Some enhancements can be incorporated into the PENELOPE code to improve its results in the nanodosimetry field. PMID- 19292003 TI - An efficient pre-object collimator based on an x-ray lens. AB - A multiprism lens (MPL) is a refractive x-ray lens with one-dimensional focusing properties. If used as a pre-object collimator in a scanning system for medical x ray imaging, it reduces the divergence of the radiation and improves on photon economy compared to a slit collimator. Potential advantages include shorter acquisition times, a reduced tube loading, or improved resolution. We present the first images acquired with a MPL in a prototype for a scanning mammography system. The lens showed a gain of flux of 1.32 compared to a slit collimator at equal resolution, or a gain in resolution of 1.31-1.44 at equal flux. We expect the gain of flux in a clinical setup with an optimized MPL and a custom-made absorption filter to reach 1.67, or 1.45-1.54 gain in resolution. PMID- 19292004 TI - Range and modulation dependencies for proton beam dose per monitor unit calculations. AB - Calculations of dose per monitor unit (D/MU) are required in addition to measurements to increase patient safety in the clinical practice of proton radiotherapy. As in conventional photon and electron therapy, the D/MU depends on several factors. This study focused on obtaining range and modulation dependence factors used in D/MU calculations for the double scattered proton beam line at the Midwest Proton Radiotherapy Institute. Three dependencies on range and one dependency on modulation were found. A carefully selected set of measurements was performed to discern these individual dependencies. Dependencies on range were due to: (1) the stopping power of the protons passing through the monitor chamber; (2) the reduction of proton fluence due to nuclear interactions within the patient; and (3) the variation of proton fluence passing through the monitor chamber due to different source-to-axis distances (SADs) for different beam ranges. Different SADs are produced by reconfigurations of beamline elements to provide different field sizes and ranges. The SAD effect on the D/MU varies smoothly as the beam range is varied, except at the beam range for which the first scatterers are exchanged and relocated to accommodate low and high beam ranges. A geometry factor was devised to model the SAD variation effect on the D/MU. The measured D/MU variation as a function of range can be predicted within 1% using the three modeled dependencies on range. Investigation of modulated beams showed that an analytical formula can predict the D/MU dependency as a function of modulation to within 1.5%. Special attention must be applied when measuring the D/MU dependence on modulation to avoid interplay between range and SAD effects. PMID- 19292005 TI - Design and characterization of a novel indicator dosimeter for blue-light radiation. AB - Optical absorption and fluorescence measurements have been done on poly[2-methoxy 5-(2'-ethyl-hexyloxy)-1,4-phenylene vinylene]/[aluminum-tris(8-hydroxyquinoline)] solutions. The authors show that there is a visible response that covers the electronic absorption of bilirubin (350-500 nm), and hence, this material is applicable for managing the radiation doses planning before treatment of jaundice of neonates, which is one of the most common reasons of hospital readmission of newborns infants. The results show that the material presents a gradation of color from orange to yellow clearly, while its peak position emission shifts from orange-red (lambda(max) = 571 nm) to green (lambda(max) = 540 nm) with the radiation exposure time. The rate of these changes can be altered by manipulations of organic solution concentration and they are usually slow (from 2 to 8 h), suggesting these color and emission changes can be used to design an easy to make, easy to read, easy to operate, low cost (< US $0.50) and accuracy for individual monitoring indicator dosimeter in order to represent easily the radiation exposure time usually used in management of neonatal jaundice. PMID- 19292006 TI - Microscintigraphy with high resolution collimators and radiographic detectors. AB - The potential use of high resolution collimators with standard radiographic detectors in place of conventional gamma cameras for high resolution microscintigraphy is presented. Polycapillary multiple hole collimators are shown to provide 10-100 micron scale spatial resolution. A series of images from arrays of 125I brachytherapy seeds in Lucite phantoms display resolution better than 0.1 mm with good sensitivity and a 30 mm field of view. In addition to application to brachytherapy seed localization, such "cellular" level resolution is necessary for high-resolution in vivo imaging in mouse models. The system could also enable the use of a wider variety of isotopes, including much lower photon energy isotopes in nuclear medicine, as the high resolution collimator allows more flexibility in detector constraints. PMID- 19292007 TI - Investigation of PVA cryogel Young's modulus stability with time, controlled by a simple reliable technique. AB - We describe a quasistatic method for mechanical characterization of tissue mimicking material used in elastography. We demonstrate that it is possible to assess the elasticity modulus with a reasonable reproducibility using simple and easy tools and methods. Possessing a simple relevant technique with evaluated relative error to assess Young's modulus of these phantoms could deeply improve the quality of the research in the field of elastography. The method was tested and validated with four samples of polyvinyl alcohol (PVA) cryogel with different elasticity values corresponding to those of stiffer soft biological tissues. Young's moduli, varying from 70 to 180 kPa depending on the number of freeze-thaw cycles (two to five), were measured within strict measurement conditions and found to have a reproducibility varying from 4% to 8%. Relative error, estimated as the ratio between observed and reference values, varied from 16% to 32%. Besides, measurement stability over 4 months was evaluated. The method demonstrated good feasibility and acceptable reproducibility for mechanically characterizing and controlled over time phantoms used for validating new potential ultrasound imaging techniques in the field of elastography. Nevertheless, in this study, investigation was performed on gel possessing young's modulus values ranging from 80 to 215 kPa. Some tissue values of Young'modulus were reported to be lower, ranging from 0.6 to 28 kPa as liver or glandular values. Consequently, further validation of this static method for mechanical characterization of phantom gels should be performed using softer PVA cryogel. PMID- 19292008 TI - A new single acquisition, two-image difference method for determining MR image SNR. AB - A new method for computing the signal-to-noise ratio (SNR) of magnetic resonance images is presented. The proposed method is a "difference of images" based technique where two images are produced from one acquisition in which the readout direction field of view (FOV) and matrix size are doubled compared to the phase encode direction. Two "normal" unaliased FOV images are produced by splitting (undersampling) the even versus odd data points in the read direction into two separate raw data sets. After image reconstruction, conventional difference of images SNR computations are applied. [Signal defined as mean within signal producing region of interest (ROI) in one image, noise defined as standard deviation of the difference between the two images using the same signal ROI position and size, divided by sqrt(2) to account for the subtraction process.] This method combines the desirable minimal acquisition time of a single image acquisition technique and the superior noise quantification characteristics of the difference of images methodology. The proposed method is more robust against system drift than existing SNR difference of images methods because the two images are effectively acquired nearly simultaneously in time. This method is compatible with phased array coils and is useful for parallel image reconstruction analysis because it is very stable. This method produces results that can be made equivalent to, and compared with, other existing SNR methods with simple known scale factors, assuming the image noise follows theoretical expectations. PMID- 19292009 TI - Age-related differences in reliance behavior attributable to costs within a human decision aid system. AB - OBJECTIVE: An empirical investigation was done to determine if there are age related differences attributable to costs in reliance on a decision aid. BACKGROUND: Costs of reliance on a decision aid may affect reliance on the aid. Older and younger adults may not perceive and respond to a dynamic cost structure equally or objectively. METHOD: Sixteen older adults (65-74 years) and 16 younger adults (18-28 years) performed a counting task with an imperfect decision aid. Two types of costs were manipulated: (a) cost of error (CoE) and (b) cost of verification (CoV). The percentage of trials in which participants agreed with the decision aid and did not perform the task manually was recorded as reliance. RESULTS: Participants decreased their reliance as the CoE increased and increased their reliance with a lower CoV; however, they tended to underrely on the decision aid. Younger adults tended to change their reliance behavior more than older adults did with the changing cost structure. CONCLUSIONS: Older and younger adults appear to interpret costs differently, with older adults being less responsive to changes in costs. Older adults may have been less able to monitor the changing costs and hence not adapt to them as well as younger adults. APPLICATION: Designers of decision aids should consider explicitly stating costs associated with reliance on the aid, as individuals may differ in how they interpret and respond to changing costs. PMID- 19292010 TI - Expertise differences in attentional strategies related to pilot decision making. AB - OBJECTIVE: We investigated expertise differences in pilot decision making by examining a hypothesized attention-action link. During simulated flight we measured the accuracy and latency of more and less expert pilots' decision outcomes and used eye tracking to measure their attention. We also examined whether decision outcomes and attentional strategies were influenced by properties of the cues indicating problems. BACKGROUND: Errors in decision making contribute to many accidents and incidents, especially among inexperienced pilots. Although much is known about decision errors in terms of their outcomes, less is known about the cognitive processes that underlie expert differences in aviation decision making. METHOD: Fourteen more expert and 14 less expert pilots flew 16 flights in a simulator. Half of the flights contained a failure requiring diagnosis and action in response to the failure. Cues signaling the failures varied in how diagnostic and/or correlated they were. RESULTS: The more expert pilots generally made better decisions in terms of speed and accuracy. Both groups made faster correct decisions when cues were high versus low in diagnosticity. Only the more expert pilots made faster correct decisions when cues were correlated. More attention was allocated to relevant cues (measured by percentage dwell time on areas of interest) when a failure was present, primarily among expert pilots. Moreover, the amount of attention to cues was associated with decision accuracy. CONCLUSION: The findings support the link between greater attention and more effective decision making. APPLICATION: The expert advantage in attention underlying decision outcomes may provide targets for improving pilot training. PMID- 19292011 TI - Perceptual-motor performance and associated kinematics in space. AB - OBJECTIVE: To determine whether the impairment of perceptual-motor performance often observed during the initial stages of space flight is attributable to the direct effects of microgravity on sensory-motor input-output relationships or to cognitive overload arising from the variety of stressors encountered in this environment. BACKGROUND: Experimental evidence is contradictory, and the present experiment investigated the role of two variables: use of ajoystick with tracking or aiming tasks and degree of arm restraint. METHODS: Five well-trained astronauts performed Fitts' reciprocal aiming task on three occasions (preflight, on the International Space Station, and postflight) under the following conditions: (a) with a stylus or ajoystick, (b) with the arm restrained or unrestrained, and (c) as a single task or together with auditory reaction time (RT) as a dual task. In addition to the speed and accuracy of responses, kinematic measures were derived from the joystick. RESULTS: In space, the slope of Fitts' function increased only in the dual task condition with the joystick. RT was also slowed in the dual condition, and there was an increase in aiming and RT errors. Percentage time to peak acceleration, velocity, and deceleration were increased when the aiming task was performed alone. CONCLUSION: These results support the cognitive overload rather than the microgravity hypothesis and indicate that an impairment in sensory-motor performance is not a necessary concomitant of space flight. APPLICATION: Impairments in perceptual-motor performance in space can be eliminated or mitigated by appropriate training and task design. PMID- 19292012 TI - Effects of simulator practice and real-world experience on cell-phone-related driver distraction. AB - OBJECTIVE: Our research examined the effects of practice on cell-phone-related driver distraction. BACKGROUND: The driving literature is ambiguous as to whether practice can reduce driver distraction from concurrent cell phone conversation. METHODS: Drivers reporting either high or low real-world cell phone usage were selected to participate in four 90-min simulated driving sessions on successive days. The research consisted of two phases: a practice phase and a novel transfer phase. RESULTS: Dual-task performance deficits persisted through practice and transfer driving conditions. Moreover, groups reporting high and low real-world experience exhibited similar driving impairments when conversing on a hands-free cell phone. CONCLUSIONS: These data indicate that practice is unlikely to eliminate the disruptive effects of concurrent cell phone use on driving. APPLICATION: Multiple regulatory agencies have considered, or are currently considering, legislation to restrict in-vehicle cell phone use. Findings reported herein may be useful to inform these public policy decisions. PMID- 19292013 TI - Does team training improve team performance? A meta-analysis. AB - OBJECTIVE: This research effort leveraged the science of training to guide a taxonomic integration and a series of meta-analyses to gauge the effectiveness and boundary conditions of team training interventions for enhancing team outcomes. BACKGROUND: Disparate effect sizes across primary studies have made it difficult to determine the true strength of the relationships between team training techniques and team outcomes. METHOD: Several meta-analytic integrations were conducted to examine the relationships between team training interventions and team functioning. Specifically, we assessed the relative effectiveness of these interventions on team cognitive, affective, process, and performance outcomes. Training content, team membership stability, and team size were investigated as potential moderators of the relationship between team training and outcomes. In total, the database consisted of 93 effect sizes representing 2650 teams. RESULTS: The results suggested that moderate, positive relationships exist between team training interventions and each of the outcome types. The findings of moderator analyses indicated that training content, team membership stability, and team size moderate the effectiveness of these interventions. CONCLUSION: Our findings suggest that team training interventions are a viable approach organizations can take in order to enhance team outcomes. They are useful for improving cognitive outcomes, affective outcomes, teamwork processes, and performance outcomes. Moreover, results suggest that training content, team membership stability, and team size moderate the effectiveness of team training interventions. APPLICATION: Applications of the results from this research are numerous. Those who design and administer training can benefit from these findings in order to improve the effectiveness of their team training interventions. PMID- 19292014 TI - [The FINESSE trial]. PMID- 19292015 TI - [Evaluation of "complex syncope: what are the indications for second-level investigations?]. AB - Syncope is a common symptom accounting for 1.1% of all admissions to the emergency department in Italy. Diagnostic and therapeutic management of patients with syncope may be complex and with a major impact on health expenditure. A standardized approach to syncope may reduce diagnostic tests, hospitalizations and health costs. After the initial "gold standard" evaluation, which includes history, physical examination, orthostatic hypotension test and ECG, several diagnostic pathways can be followed. It has been shown that a correct initial evaluation and a thorough knowledge of syncope can reduce needless testing and increase diagnostic yield, optimizing resource management. In this review we aim to underscore the key points of the management of patients with syncope and the main indications for specific second-level examinations, such as those for neuroautonomic evaluation (tilt table test, carotid sinus massage) and implantable loop recorder. The role of Syncope Units in the management of patients with temporary loss of consciousness is also described. PMID- 19292016 TI - [Alcohol-induced cardiac disease]. AB - Excessive and chronic ethanol consumption exerts deleterious and diffused effects on the myocardium, independent of coronary atherosclerosis, arterial hypertension, valvular disease or congenital heart disease. Although the effects of chronic alcoholism on systolic cardiac function are well known, diastolic involvement has been evaluated only partially. This short review summarizes the experimental and clinical evidence for alcohol-induced cardiac disease. PMID- 19292017 TI - [Mechanisms of myocardial cell protection from ischemia/reperfusion injury and potential clinical implications]. AB - Myocardial cell damage caused by myocardial ischemia results from several factors that include the duration of ischemia, oxygen demand by cardiomyocytes at the time of ischemia, and the presence and entity of collateral blood flow to the ischemic area. Importantly, myocardial cell injury may derive not only from ischemia itself but also from detrimental phenomena occurring during the restoration of myocardial blood flow after the ischemic episode (reperfusion damage). In the last decades a lot of studies have demonstrated that cardiomyocytes have several mechanisms that provide them protection against the damage deriving from ischemia-reperfusion, also allowing a prolongation of survival in the most severe cases. In this article we review some of these mechanisms, also discussing their present and/or potential therapeutic applications in the clinical setting. The topics include the interventions aimed at reducing cardiac work through a reduction of oxygen demand by myocardial cells or at optimizing the utilization of energetic resources by myocardial cells in situations of ischemia, the importance of phenomena such as ischemic preconditioning (early and delayed) and postconditioning of myocardial cells, and, finally, the theoretic possibility of interventions aimed at preventing cell death consequent to apoptosis. PMID- 19292018 TI - [CardiReset: general medicine and monitoring of cardiovascular risk factors in a whole region]. AB - BACKGROUND: The CardioRESET project was carried out in 2005 to evaluate the feasibility of a cardiovascular risk factor survey by general practitioners in Friuli Venezia Giulia, a north-eastern Italian region. METHODS: We randomized 2701 subjects (1336 males and 1365 females), aged 35-74 years, from the general population. The family doctors surveyed the randomized population sample using the standardized methods of the Osservatorio Epidemiologico Cardiovascolare, a reference national survey. RESULTS: The participation rate was 85.4% and all variables were recorded at least in 60% of subjects. Mean values of risk factors, prevalence of hypertension, dyslipidemia, diabetes and data on their control, smoking habits were comparable with data from the Osservatorio Epidemiologico Cardiovascolare; only the mean value of low-density lipoprotein cholesterolemia was higher in our region. CONCLUSIONS: This experience points out that in a small region it is possible to engage general practitioners to achieve a standardized surveillance of cardiovascular risk factors at a low cost. PMID- 19292019 TI - [Surgical repair of a giant left ventricular aneurysm]. PMID- 19292020 TI - [Chest pain evaluation project]. AB - The evaluation of acute chest pain remains challenging, despite many insights and innovations over the past two decades. The percentage of patients presenting at the emergency department with acute chest pain who are subsequently admitted to the hospital appears to be increasing. Patients with acute coronary syndromes who are inadvertently discharged from the emergency department have an adverse short term prognosis. However, the admission of a patient with chest pain who is at low risk for acute coronary syndrome can lead to unnecessary tests and procedures, with their burden of costs and complications. Therefore, with increasing economic pressures on health care, physicians and administrators are interested in improving the efficiency of care for patients with acute chest pain. Since the emergency department organization (i.e. the availability of an intensive observational area) and integration of care and treatment between emergency physicians and cardiologists greatly differ over the national territory, the purpose of the present position paper is two-fold: first, to review the evidence based efficacy and utility of various diagnostic tools, and, second, to delineate the basic critical pathways (describing key steps for care and treatment) that need to be implemented in order to standardize and expedite the evaluation of chest pain patients, making their diagnosis and treatment as uniform as possible across the country. PMID- 19292021 TI - [Treatment of acute infarction in the province of Foggia: primary angioplasty exists, too]. PMID- 19292022 TI - Preventing errors in patient management: the emergency department clinician and the toxicology laboratory. AB - Poor communication between the clinical toxicologist and the toxicology laboratory can result in a variety of problems. Clinicians must familiarize themselves with the toxicology assays that are available in their hospital and how to interpret assay results. Toxicology tests ordered in the emergency department should provide useful information for patient management and disposition. Toxicology laboratory personnel should have a good working relationship with emergency department clinicians and clinical toxicologists to maximize the usefulness of the laboratory in patient management. PMID- 19292023 TI - Hepatotoxicity in an infant following supratherapeutic dosing of acetaminophen for twenty-four hours. AB - Severe hepatotoxicity in acute acetaminophen overdose in children is uncommon. This apparent protection does not extend to chronic or repeated high dose acetaminophen administration. The Illinois Poison Center (IPC) was consulted on a case involving a 7 month old 17 lb infant, who had presented the day prior to an emergency department with complaint of febrile illness. Patient had been prescribed acetaminophen for symptoms, and the mother called IPC when she realized that she had been incorrectly giving the patient concentrated infant acetaminophen (80 mg per 0.8 mL) totalling 42.3 mg/kg/dose or 234 mg/kg/24 hours instead of children's acetaminophen (160 mg/5 ml). Per recently published national triage guidelines, the patient was referred to an acute care facility for evaluation. The patient presented with significantly elevated aspartate aminotransferrase (AST) and alanine aminotransferrase (ALT) levels of 1339 and 907 U/L respectively, and was initiated on i.v. n-acetylcysteine therapy. The patient responded well to therapy, with AST and ALT declining to 145 and 479 U/L respectively over the next 48 hours. This case represents one of the shortest durations of chronic acetaminophen therapy to cause hepatic injury in an infant. Further, while the level was only slightly above the national triage guideline recommended referral dose per 24 hours, significant hepatotoxicity was observed. PMID- 19292024 TI - Cocaine: history, social implications, and toxicity: a review. AB - The amount of positive cocaine results in an urban emergency department are staggering. The ages of use are becoming more common in older age groups. Most of these patients have underlying medical conditions, including end-stage renal disease (on hemodialysis) and heart and lung disease. Most of their visits to the emergency department are for cocaine exacerbation of underlying chronic condition, adding exponentially to health care dollars. This article describes the history and pharmacology of illicit cocaine use. PMID- 19292025 TI - A review of the use of ethyl glucuronide as a marker for ethanol consumption in forensic and clinical medicine. AB - Ethyl glucuronide (EtG) is a direct phase-II metabolite of ethanol formed through the UDP-glucuronosyl transferase catalyzed conjugation of ethanol with glucuronic acid. It has been detected in many antemortem and postmortem biological matrices using a variety of analytical methods. Due to its long urinary elimination time, detectability in hair, specificity for ethanol exposure, and low detection limits of assays, the use of EtG has been proposed as a marker of recent ethanol intake in a variety of clinical and legal settings, including medical monitoring for relapse, emergency department patient evaluation, postmortem assessments, and transportation accident investigation. However, challenges associated with factors such as establishing appropriate cut-off levels capable of distinguishing between drinking and nonbeverage sources of ethanol exposure, nonuniform laboratory reporting limits, sample stability, and microbial activity substantially complicate accurate interpretation of results. The following review briefly explores the history, utility, and limitations of EtG in contemporary medical and forensic practice. PMID- 19292026 TI - Clinical approach to clinical herbal toxicity. AB - This article is a review of the history and current status of herbs in the United States. The clinical effects of herbs and herbal medicine will be reviewed with respect to side effects and clinical toxicity. Treatment of herbal toxicity will be described where indicated. PMID- 19292027 TI - The effect of complimentary and alternative medicine products on laboratory testing. AB - A multi-billion dollar industry has evolved over the last decade based on herbal product sales with an underlying belief that herbals are natural and therefore safe. The herbal product industry is essentially unregulated and producers are not required to follow good manufacturing practices (GMP). Batch to batch product variation, heavy metal and pesticide contamination, and even therapeutic drug contamination are problematic. Compounding these manufacturing issues are drug to drug and drug to herbal interactions that can cause cytochrome induction or inhibition. It is important for physicians to query their patients on herbal use and educate them on the potential adverse reactions. Herbals have been used for thousands of years and undoubtedly have demonstrated health benefits. However, more research is needed to gain an understanding of the complexity issues from mechanism of action to interference with clinical laboratory testing. PMID- 19292028 TI - Laboratory interferences with the newer cyanide antidote: hydroxocobalamin. AB - Cyanide poisoning occurs in many smoke inhalation victims. The newest FDA approved treatment for acute cyanide intoxication is hydroxocobalamin (Cyanokit). However, hydroxocobalamin exhibits chemical properties that can disrupt several clinical laboratory tests. Knowledge of these effects on laboratory tests can be useful in assisting laboratory technicians and clinicians in managing these patients. This article briefly discusses acute cyanide poisoning and treatment, and summarizes laboratory interferences that have been reported with the use of hydroxocobalamin. PMID- 19292029 TI - Normal reference value of red blood cell count of Chinese young men and geographical factors. AB - AIM: This paper aims at supplying a scientific basis for unifying the normal reference value standard of red blood cell count of Chinese young men. METHODS: The research is made to study the relationship between the normal reference value of 41,759 examples of red blood cell count of young men and 8 geographical factors in 334 areas in China, which is determined by the microscopical counting method. RESULTS: It is found that the correlation of geographical factors and the normal reference value of red blood cell count of young men is quite significant (F = 224.98, P = 0.000). By using the method of stepwise regression analysis, one regression equation is inferred. CONCLUSION: If geographical values are obtained in a certain area, the normal reference value of red blood cell count of young men in this area can be reckoned by using the regression equation. Furthermore, according to the geographical factors, China can be divided into eight districts: Northeast China District, North China District, Shanxi-Shaanxi-Inner Mongolia District, Middle and Lower reaches of the Changjiang River District, Southeast China District, Northwest China District, Southwest China District, and Qinghai Tibet Plateau District. PMID- 19292030 TI - Death by polonium-210: lessons learned from the murder of former Soviet spy Alexander Litvinenko. AB - The medical response to radiation--whether the result of radiological warfare, terrorist deployment of improvised radiation dispersal weapons, political assassination, occupational or industrial accidents or the medically radiated patient remains one of the least taught among all disciplines within medical education. In the aftermath of 9/11 among medical vulnerabilities to toxicant threats, of all the categories of weapons of mass destruction (WMD)--whether using the CBRNE (chemical, biological, radiological, nuclear, explosive) or NBC (nuclear, biological, chemical) acronym--radiation is the least taught in professional schools, responder cultures or civil preparedness organizations. To date, few health care professionals (HCP) possess the fundamental knowledge or skills to identify and diagnose, let alone treat a radiation victim; this vulnerability made even more obvious in the aftermath of the high profile assassination of former Russian agent Alexander Litvinenko. He was poisoned with Polonium210. Radioactive substances are ubiquitous with radiation sources being in or transported through virtually every region nationwide. It is essential to increase preparedness among community and rural health care facilities as well as urban and university hospitals. Managing radiation injuries effectively requires access to specialized equipment and expertise. Radiation sickness is progressive and may require acute, critical and long-term care throughout the course of illness. Regardless of the source, preparedness rests upon acknowledging a threat exists and dedicating the resources to address the risks including the enhancement of training and equipment. Mass or individual exposures to radiation present unique challenges to the entire response continuum from law enforcement, first responders and emergency medical care. Increased education about and practice in responding to radiological threats is essential to enhance preparedness. PMID- 19292031 TI - American Academy of Clinical Toxicology: "question of the day". PMID- 19292032 TI - [Cloning, expression, purification and identification of multi-potent transcription factor CTCF recombinant and its segments]. AB - OBJECTIVE: To clone ctc f cDNA, N, Zn and C fragments separately into expresstion vector, purify and identify the expressed proteins. METHODS: Using the recombinant plasmid pGEM7Zf (-)-ctc f as template for PCR, pGEX-4T-2-ctc f, pGEX 4T-2-ctc f-N, pGEX-4T-2- ctc f-Zn and pGEX-4T-2- ctc f-C recombinants were constructed successfully. After transformed into E. coli BL21 cells, the recombinants were confirmed by enzyme digestion and sequence analysis. After optimizing the IPTG inducing condition, the purified GST fusion proteins with affinity chromatography were conformed by Far-Western blotting. RESULTS: The recombinant plasmids pGEX-4T-2-ctc f, pGEX-4T-2- ctc f-N, pGEX-4T-2-ctc f-Zn and pGEX-4T-2-ctc f-C were confirmed by restriction enzyme assay and sequencing. All GST fusion proteins, CTCF, CTCF-N, CTCF-Zn and CTCF-C were successfully expressed at the optimal parameters and purified with affinity chromatography, and specifically recognized by anti-GST antibody. CONCLUSION: Ctc f, ctc f-n, ctc f Zn and ctc f-c gene recombinants were constructed successfully and their corresponding fusion proteins were successfully purified with affinity chromatography and identified. PMID- 19292033 TI - [Effects of ARLTS1 gene on growth and apoptosis of epithelial ovarian cancer SKOV3 cells]. AB - OBJECTIVE: To investigate the effect of ARLTS1 (ADP-ribosylation factor-like tumor suppressor gene 1) on the growth and apoptosis of ovarian cystadenocarcinoma cell line SKOV3. METHODS: The recombinant plasmid pCMV-Tag 3B ARLTS1 was constructed and transfected into SKOV3 cells by liposome protocol. The expression of ARLTS1 mRNA and its protein were examined by RT-PCR and Western blotting. Effects of ARLTS1 gene on growth and apoptosis of the transfected cells were evaluated by MTT assay and flow cytometry (FCM). The alterations of caspase 3 and bcl-2 protein levels were examined by Western blotting. RESULTS: The stable transfection of pCMV-Tag 3B-ARLTS1 in SKOV3 cells were obtained and verified by RT-PCR and Western blotting methods. After transfected with ARLTS1, more SKOV3 cells were observed in S phase by FCM compared with those in the other two groups (P is 0.035 and 0.011, respectively). The 36.7% apoptotic index of ARLTS1 transfected cell was significantly higher than that of the other two groups (P < 0.001). The growth of ARLTS1 transfected cells was dramatically inhibited compared with cells transfected with Vector (P < 0.05). Western blotting indicated a significantly decrease in caspase-3 and bcl-2 protein levels in cells transfected with ARLTS1 compared with SKOV3 cells (P is 0.021 and 0.013, respectively). CONCLUSION: ARLTS1 transfection can inhibit proliferation and induce apoptosis of SKOV3 cells in vitro and down-regulate the expression of caspase-3 and bcl-2. ARLTS1 gene may be a candidate tumor suppressor gene in ovarian cancer. PMID- 19292034 TI - [Oxidative damage of gasoline engine exhausts to rat lung tissues]. AB - OBJECTIVE: To study the effects of extracts of condensate, particulates and semivolatile organic compounds from gasoline engine exhaust on DNA damage, 8 oxoguanine DNA glycosylase-1 (OGG1) expression, and changes of ultra-structures in lungs of rats. METHODS: Organic extracts of gasoline engine exhaust (GEE) was intratrachealy instilled into rat lungs at 0, 5.6, 16.7, and 50.0 L/kg body weight, respectively, once a week for a month. The single DNA strand break was measured by comet assay. The OGG1 was determined using immunohistochemistry method. The ultrastructure of lung cells was observed with electronic microscope. RESULTS: The rates of tailed cells detected by the comet assay increased significantly when the rats were exposed to 16.7 and 50.0 L/kg of GEE compared with those exposed to solvent only (P < 0.05). However, the tail length did not differ significantly between the groups. Similarly, exposure to 16.7 and 50.0 L/kg of GEE led to increased OGG1 significantly. Significant changes of mitochondria in type I and II alveolar cells as well as respiratory bronchiole epithelial cells were observed, which included decrease of numbers, pyknosis and swelling. CONCLUSION: Gasoline engine exhausts induce single DNA strand break, increase OGG1 expression, decrease numbers of mitochondria, and destroy ultrastructures of mitochondria in various lung cells of rats. PMID- 19292035 TI - [Effects of prepubertal exposure to estradiol benzoate on testicular development and function of Sprague-Dawley rats]. AB - OBJECTIVE: To investigate the effects of prepubertal exposure to estradiol benzoate (EB) on the testicular development and function of SD rats. METHODS: Fifty four 21-day-old male SD rats were randomly and equally divided into 2 experimental groups (AEa and AEb, n = 18 each) and one control group (AC, n = 18). The rats in the experimental groups were injected (s. c.) with EB dissolved in corn oil at a dose of 0.1 (AEa) and 100.0 microg/(kg x d) (AEb) respectively during prepuberty for 14 d [from postnatal day (PND) 22 to PND 35]. The rats in the control group received vehicle injections only. The testicular development of the rats was observed. The testes were harvested at the stages of late puberty (PND 50), sexual maturity (PND 64) and adulthood (PND 130) (n = 6 at each stage). The serum testosterone, the weights and histology of the testes, and the quality of sperms in the epididymal cauda were examined. RESULTS: The testes of rats in group AEb descended at day 43.17 +/- 1.72, later than the rats in the AC group (27.00 +/- 0.89, P < 0.01). At the stage of late puberty, the AEb group had lower serum testosterone than the AC group (P < 0.05) and had less unilateral testes weights than the AC group (P < 0.01). Compared with the AC group, the histological alteration of the testes of rats in the AEb group included seminiferous tubules maldevelopment, decreased cell numbers of seminiferous epithelia, spermatogenesis blocking and Leydig cells aplasia. At the stage of sexual maturity, the AEb group had less unilateral testes weight than the AC group (P < 0.01) and maintained similar histological alterations of testes as at the stage of late puberty. At the stage of adulthood, no differences were observed among the three groups in the unilateral testes weights and the histological features of the testes (P > 0.05). However, the AEb group had significant lower density and motility of sperms and percentage of grade a+b sperms in the epididymal cauda than the AC group (P < 0.05). CONCLUSION: Prepubertal exposure to low dose of EB does not affect the testicular development and function of SD rats, while high dose [100 microg/(kg x d) x 14 d] exposure has significant short (PND 50 and 64) and long-term (PND 130) toxic effects. The toxic effects are associated with the damage of Leydig and Sertoli cells. PMID- 19292036 TI - [Expression of transferrin receptor 2 in mononuclear cells from children with acute leukemia]. AB - OBJECTIVE: To study the expression of TFR2 mRNA in mononuclear cells of bone marrow and peripheral blood from children with acute lymphoblastic leukemia, and to explore possible correlations between TFR2 expression and a panel of prognostic/risk factors. METHODS: Bone marrow or peripheral mononuclear cells were isolated from 56 children with newly diagnosed acute lymphoblastic leukemia (ALL) and 15 normal children as control. TFR2 mRNA expression level in mononuclear cell was determined by real-time fluorescence quantitative RT-PCR. RESULTS: Relative expression level of TFR2 mRNA in prednisone good responders (median: 0.0848) was significantly higher than that in prednisone poor responders (median: 0.0126) (P = 0.038). The medians of TFR2 mRNA expression levels in low risk, moderate-risk and high-risk ALL were 0.1677, 0.0728 and 0.0125 respectively (P = 0.003). The medians of TFR2 mRNA expression levels in three ALL groups with initial WBC counts less than 50 x 10(9)/L, from 50 x 10(9)/L to 100 x 10(9)/L and more than 100 x 10(9)/L were 0.0974, 0.0294 and 0.0078 (P = 0. 013). In addition, the relative TFR2 mRNA level in B-lineage ALL was significantly higher than that in T-ALL, with medians of 0.0636 and 0.0065 respectively (P = 0.004). Ranked correlation analysis revealed that TFR2 expression was negatively correlated to initial white blood cell count, percentage of blast cells and absolute numbers of blasts in peripheral blood, with ranked correlation coefficients of -0.398, 0.307 and 0.421 respectively (correponding P values were 0.003, 0.022 and 0.001). CONCLUSION: TFR2 might be a novel prognostic factor for ALL. PMID- 19292037 TI - [The effect and pathophysiological mechanism of high free fatty acids(FfAs) on the cardiac structure and function]. AB - OBJECTIVE: To study the pathophysiological links between elevated circulating FFAs concentration and the cardiac structure, and their function in obese insulin resistance rat model. METHODS: 4 weeks age male SD rats were fed with high-fat chow (OB) or standard laboratory chow (NC) respectively. Whole-body insulin sensitivity, the maximum velocity of myocardial contraction (+dp/dt(max)) and the maximum velocity of myocardial diastole (-dp/dt(max)) of intracardiac pressure, and myocardiac cell diameter (MCD) were measured. The concentrations of triglyceride (TG), FFAs and angiotensin II (Ang II) both in blood and in left ventricular portions of heart and the expressions of NF-kappaB, I-kappaB and iNOS in myocardium were analyzed. RESULTS: OB group developed obesity and left ventricular hypertrophy, and their insulin sensitivity was much lower than that of control group. Obese rats had higher plasma concentrations of TG, FFAs and Ang II. Accordingly, dramatic lipid deposition occurred within cardiomyocytes of obese rats, and the value of myocardiac Ang II was also increased. High-fat diet also induced a progressive decrease in values of +dp/dt(max) and -dp/dt(max). The higher expressions of NF-kappaB and iNOS in myocardium were observed in OB group, while IkappaB lower. Intramyocardial lipid deposition was associated with plasma FFAs concentrations (r = 0.80, P < 0.01). Intramyocardial FFAs concentration was associated with myocardial Ang II concentration (r = 0.74, P < 0.05) and changes in expressions of NF-kappaB (r = 0.86, P < 0.01), iNOS (r = 0.66, P < 0.05). The contractile dysfunction was associated with intramyocardial lipid deposition (r = -0.87, P < 0.01), Ang II (r = -0.52, P < 0.05) and expressions of NF-kappaB (r = 0.57, P < 0. 01), iNOS (r = -0.70, P < 0. 01). The diastolic dysfunction was associated with intramyocardial lipid deposition ( r = -0.85, P < 0.01), Ang II (r = -0.82, P<0.01) and expressions of NF-kappaB (r = -0.75, P < 0.01), iNOS (r = -0.78, P < 0.01). CONCLUSION: In obese/insulin resistance, state ectopic lipid accumulation in myocardium as the results of elevated circulating FFAs and TG concentration impairs cardiac systolic and diastolic functions. It is logical to deduce that ectopic lipid accumulation in myocardium may increase RAS activity and expressions of NF-kappaB, iNOS in myocardium, all of them have important roles to increase the risk of congestive heart failure in obese subjects. PMID- 19292038 TI - [Construction and property study of recombinant Lactococcus lactis with non fusion expressing of beta-galactosidase]. AB - OBJECTIVE: To construct recombinant Lactococcus lactis strains exhibiting high beta-galactosidase activity in non-fusion way, and study their enzyme activities and enzyme secretion rates. METHODS: The recombinant plasmids pMG36e-lacZ 1.1480 and pMG36e-lacZ wch9901 which could express beta-galactosidase from Lactobacillus delbrueckii subsp. bulgaricus in non-fusion way in Escherichia coli were obtained and transformed into Lactococcus lactis subsp. lactis MG1363. The beta galactosidase activity of resulting recombinant L. lactis in different incubation periods and lactose concentrations, and their enzyme secretion rates in different culture conditions were examined. RESULTS: Recombinant L. lactis carrying pMG36e lacZ wch9901 (MG1363/pMG36e-lacZ wch9901) exhibited the highest beta galactosidase activity. Its enzyme activity was (16.95 +/- 0.09) U/mg pro, which was 2.75 folds of that of the native counterpart; recombinant L. lactis reached its enzyme producing peak after grown for 24 h; decreased enzyme activity of recombinant L. lactis were observed when incubated in medium containing lactose; the beta-galactosidase expressed by recombinant strains could be secreted into the culture medium, and the highest secretion rate (27.09 +/- 0.05)% was observed when the culture medium contained 20 g/L of lactose and without erythromycin. CONCLUSION: High level expression of non-fusion beta-galactosidase with secretion in recombinant L. lactis strains was obtained. This will be very helpful for the further developing of live delivery bacteria of beta-galactosidase. PMID- 19292039 TI - [Detecting int I 1 gene expression of Pseudomonas aeruginosa with fluorescent quantitative PCR assay]. AB - OBJECTIVE: In order to understand the role of integron, fluorescent quantitative polymerase chain reaction (FQ-PCR)was developed to measure the changes in int I 1 gene expression of Pseudomonas Aeruginosa in biofilm and planktonic cells. METHODS: Three clinical strains of P. aeruginosa with int I 1 gene (SW07, R07 and TH12) were cultured in planktonic cells and biofilm cells. The total RNA of these cultured bacteria were extracted by the conventional method. The FQ-PCR was developed to measure the changes in int I 1 mRNA expression of the P. aeruginosa with bacterial 16s rRNA as an internal control. RESULTS: The three clinical strains of P. aeruginosa expressed int I 1 mRNA in both biofilm and planktonic cells, but with different levels. The int 1 mRNA expressed by the RO7, SW07 and TH12 strains in the biofilm cells were 1.4, 5.7 and 128 times higher than in the planktonic cells, respectively. CONCLUSION: The int I 1 gene expression of P. aeruginosa in the biofilm is up-regulated at mRNA level. The integron may capture and accumulate drug resistance gene cassettes more effectively in the biofilm condition. PMID- 19292040 TI - [Hypoxia inducible factor-1alpha expression trends in hippocampus of rat of different ages]. AB - OBJECTIVE: To reveal the expression trends of HIF-1alpha in hippocampus of different age rats, investigate the role of HIF-1alpha in the aging process of nervous system. METHODS: The cerebral tissues was collectd from rats of different age, including 3, 18, 24, 36 months old. There were 6 rats in each age group. The expressions of nissl body and HIF-1alpha in different part of hippocampus were observed by nissl staining and immunohistochemical technique. RESULTS: (1) With the increase of rat age, nerve cells appeared to be bigger and to arrange sparsely, while the nissl body decreased; (2) The positive HIF-1alpha staining cells in CA1, CA3 region of hippocampus increased along with the increase of rat age. The difference between any two age groups showed statistical significance (P < 0.05). CONCLUSION: The function of protein synthesis weakened in nerve cells but the expression of HIF-1alpha increased with the age increasing, which may play an important role in aging of nervous system. PMID- 19292041 TI - [Expression of p38MAPK in the olfactory bulb of rats with Alzheimer's disease]. AB - OBJECTIVE: To identify the changes of expression of p38MAPK in the olfactory bulb (OB) of rats with Alzheimer's Disease (AD) and to explore the functional mechanism of p38MAPK in the pathological changes of AD. METHODS: AD animal model was established by injecting amyloid-beta peptide 25-35 into the lateral cerebral ventricle of rats. The learning and memory abilities of rats were measured by Y maze experiment. The expressions of p38MAPK in the OBs of rats in the AD group, saline control group, p38MAPK inhibitor group and inhibitor control (con inhibitor) group were examined by immunohistochemistry. RESULTS: (1) The capabilities of learning and memory of the rats were impaired significantly at day 7 and day 14 after the induction of AD. Significant learning and memory differences were found (P < 0.05) between the AD group and the other groups. (2) The expression of p38MAPK in the OBs of rats was found at day 4 after the induction of AD. The expression increased with time. The AD rats had more positive p38MAPK cells than those in the saline controls (P < 0.05). The expression of phospho-p38MAPK in the rats with AD increased significantly (P < 0.01) compared with those in the saline group. The AD rats treated with p38MAPK inhibitor had less expression of phospho-p38MAPK than those in the AD group and the inhibitor control (P < 0.01). CONCLUSIONS; Abeta25-35 can activate p38MAPK signal transduction pathway. p38MAPK may play a role in the formation of dysosmia in AD. SB203580, a p38MAPK inhibitor, can reduce the neurotoxicity evoked by p38MAPK. PMID- 19292042 TI - [Expression of glucose transporter 1 in human breast carcinoma and its clinical significance]. AB - OBJECTIVE: To investigate the expression of glucose transporter 1 (Glut1) in human breast cancer and its relationship to proliferating cell nuclear antigen (PCNA) protein, other tumor biomarkers and clinical pathologic factors. METHODS: Imunohistochemical staining (SP) was applied to measure the expression of Glut1 and PCNA in 20 cases of human breast fibroadenoma, 20 cases of usual hyperplasia and 80 cases of breast carcinoma. RESULTS: Glut1 was not found expressing in breast fibroadenoma and hyperplastic lesions. In contrast, the total positive rate of Glut1 in breast carcinoma was 58.8% (47/80); that in the ductal carcinoma in situ (DCIS) was 45.0% (9/20), that in the well-differentiated invasive carcinoma was 50.0% (15/30) and that in the poorly differentiated was 76.7% (23/30). The total positive rate of PCNA in breast carcinoma was 75% (60/80), that in DCIS was 65% (13/20) and that in invasive carcinoma was 78.3% (47/60). There was a positive correlation between Glut1 and PCNA level (r = 0.742, P (< 0.01). CONCLUSION: The overexpression of Glut1 play important roles in carcinogenesis and progression of breast carcinoma and closely correlate with cell proliferation of breast carcinoma, may suggest different therapeutic approaches or the need for closer follow-up, and be wished to become a new target for treatment of breast carcinoma. PMID- 19292043 TI - [Expression of VEGF and its significance in primary diffuse large B-cell lymphoma of the female genital system]. AB - OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) and its significance in primary diffuse large B-cell lymphoma of the female genital system. METHODS: VEGF mRNA and expression of VEGF protein were detected by real-time fluorescence quantitative PCR method and immunohistochemistry, respecitvely. Microvessel density (MVD) was obtained after staining of microvessels using CD34 antibody. The relationship between the expression of VEGF and MVD, clinical features and prognosis of primary diffuse large B-cell lymphoma of the female genital system patients were analyzed. RESULTS: (1) Over expression of VEGF mRNA was found in 17/20 (85.0%) by real-time fluorescence quantitative PCR method and the tumor cells expressed VEGF protein in 48/56 (85.7%) by immunohistochemistry; (2) The expression of VEGF was positively correlated to the MVD of the tumor (r = 0.76, P = 0.0170); (VEGF expression was correlated to clinical stages, B symptom and the serum level of lactate dehydrogenase (LDH) (r = 0.77, P = 0.044; r = 0.58, P = 0.023; r = 0.69, P = 0.017); (4) Cox multivariate analysis demonstrated that the age of more than 60 years and the positive VEGF expression were independent prognostic factors (P < 0.05). CONCLUSION: VEGF was widely expressed in the tumor cells of primary diffuse large B-cell lymphoma of the female genital system and may be related to the clinical features and prognosis of primary diffuse large B-cell lymphoma of the female genital system patients. PMID- 19292044 TI - [Relationship of 25(OH)VD with bone mass and other indicators in male patients with diabetes mellitus]. AB - OBJECTIVE: To investigate the relationship of serum 25- Hydroxy D with bone mass density and other indicators in male patients with diabetes. METHODS: Bone mass density (BMD), serum 25- HydroxyD (25 (OH)VD) and several other biochemical indicators were measured in 82 male patients with diabetes, among whom 49 had osteoporosis. RESULTS: The diabetic patients with osteoporosis had higher tartrate-resistant acid phosphatase-5b (TRCAP-5b) and lower 25(OH)VD, testosterone (T), Dehydroepiandrosterone sulfate (DHEA-S), and bone-specific alkaline phosphatase (B-ALP) than those without osteoporosis (P < 0.05). The Parathormone (PTH) and albumin-creatinine ratio (UAlb/Cr) also increased in the diabetic patients with osteoporosis, but with no statistical significance. No difference was observed in glycosylated hemoglobin (HBA1c) between the diabetic patients with and without osteoporosis. The serum 25(OH)VD was negatively correlated with the duration of diabetes in both groups (P < 0.05), which was independent from the increase of age. The serum 25(OH)VD was positively correlated with T score of neck, ward, greater trochanter of femur and vertebrae lumbales, and T and DHEA-S in both patients with and without osteoporosis (P < 0.05). The 25(OH)VD was also positively correlated with PTH in the patients with osteoporosis. Negative correlation was noted between 25(OH)VD and B-ALP. There were no correlations between 25(OH)VD and TRCAP-5b, HBA1c and UAlb/Cr. CONCLUSION: Serum 25(OH)VD, T and DHEA-S may play an important role in the development of osteoporosis in male patients with diabetes mellitus. PMID- 19292045 TI - [Expression of glomerular heparan sulfate domains in pediatric patients with minimal change nephrotic syndrome]. AB - OBJECTIVES: To investigate the expression of glomerular heparin sulfate (HS) in paediatric patients with minimal change nephritic syndrome (MCNS). METHODS: The kidyney tissues were collected by biopsy from 13 paediatric patients with MCNS, while 5 normal renal biopsy samples were used as control. HS in glomeruli was analysed by indirect immunofluorescence staining using four different monoclonal antibodies, Hepss1, 3G10, JM403 and 10E4, which all recognize distinct HS species and each interacts with a specific HS domain. The concentrations of urine heparan sulfate also were measured by enzyme-linked immunosorbent assay (Elisa). RESULTS: Expression of HS fine domains was aberrant in paediatric patients compared with control subjects. Children with MCNS in replase showed a decreased glomerular expression of 10E4, JM403 and Hepss1 (P < 0.05). The level of urinary HS was significantly increased in peadiatric patients with MCNS when compared with that in control subjects (P < 0.01). CONCLUSION: These results suggest that loss of heparan sulphate in renal tissue may play a role in the pathogenesis of MCNS proteinuria. PMID- 19292046 TI - [A proton magnetic spectroscopy research on hippocampus metabolisms in people with suicide-attempted depressions]. AB - OBJECTIVE: To investigate the metabolic features of hippocampus in people with suicide-attempted depressions. METHODS: Proton resonance spectroscopy (1H-MRS) was performed on the bilateral hippocampi of 24 patients with suicide-attempted depressions and 24 healthy people. The ratios of N-acetylasparte(NAA)/creatine (Cr), choline(Cho)/Cr and myoinositol(mI)/Cr were calculated and used as markers reflecting the metabolic levels of NAA, Cho and mI. The differences between the patients with suicide-attempted depressions and the healthy people were analyzed. The left hippocampus was also compared with the right to determine the asymmetry of the hippocampus metabolic. RESULTS: The ratio of NAA/Cr in the left hippocampus of the patients with suicide-attempted depressions was significantly less than that of the healthy control (P < 0.05). The ratio of NAA/Cr in the left hippocampus was significantly greater than in the right hippocampus of the healthy control (P < 0.05). However, the left-right deference of the ratio of NAA/Cr disappeared in the patients with suicide-attempted depressions. There was an asymmetry between the left and right hippocampi in the healthy people (P < 0.05), but not in the patients with suicide-attempted depressions (P > 0.05). No correlations between the metabolic markers and the course of disease, number of episodes and HAMD17 scores were found in the patients with suicide-attempted depressions (P > 0.05). CONCLUSION: There is a decrease of metabolite in the left hippocampus of patients with suicide-attempted depressions. The asymmetry of the bilateral hippocampi disappears in the patients with suicide-attempted depressions. PMID- 19292047 TI - [The labral plicae entrapment syndrome of hip in children: an ultrasonography study]. AB - OBJECTIVE: To study the ultrasonographic (US) features of labral plicae entrapment syndrome of hip (LPEH) in children and to evaluate the value of US in diagnosing LPEH. METHODS: (1) Twenty six LPEH models and 38 sham LPEH hips were established surgically from 32 children cadavers [15 male and 17 female, age from 2 to 8 years, mean age of (6.12 +/- 2.13) years]. US was performed on these hips double-blindly. The sensitivity, specificity, positive and negative predictive values were calculated. (2) A total of 21 children (17 male and 4 female, mean age (5.95 +/- 2.67) years) with unilateral LPEH and 21 age and gender matched children with normal hips were consecutively recruited. The 21 symptomatic hips, 21 asymptomatic hips and 42 normal hips were examined by the US (ATL 5000) using the high-frequency (7.5-12 MHz) linear array transducer. RESULTS: (1) The sensitivity, specificity, positive and negative predictive values of the US for the LPEH model were 88%, 84%, 79%, and 91%, respectively. (2) Fluids in hip joints were detected in all of the 21 symptomatic hips. No fluid was detected in the asymptomatic and normal hips (less than 2mm); The mean width of the inferomedial recess was significantly larger than that of the anterior recess (12.50 mm vs. 4.35 mm, P < 0.05) in the 21 symptomatic hip joints; The echogenic entrapped labral plicaes were demonstrated in the inferomedial recess of the 21 symptomatic hip joints, with a length ranging from 5.3 to 25.0 mm [mean(15.63 +/- 5.57) mm) and a width ranging from 4.0 to 17.0 mm [mean (8.90 +/- 7.81) mm], respectively; No color signal of blood flow was demonstrated in 90% of the 21 entrapped labral plicaes; With regard to the mean thickness of cartilage of femora head, anterior layer and posterior layer of the anterior capsule, there were no statistical significant differences between the three groups (P > 0.05). After manual reduction and conservative treatment, all of the entrapped labral plicae and fluids disappeared in the US follow-up examinations. CONCLUSION: The entrapped labral plicae in the inferomedial recess of hip joints could be visualized by the US. The US provides a useful diagnostic clue to LPEH in children. PMID- 19292048 TI - [Relationships between PON1 L55M polymorphism and coronary heart diseases complicated with fasting hyperglycemia in Han populations in Guangdong]. AB - OBJECTIVE: To investigate the relationships between Paraoxonase1 (PON1) gene L55M polymorphism and coronary heart diseases (CHD) complicated with fasting hyperglycemia (FHG) or type 2 diabetes mellitus (T2DM). METHODS: Two hundreds and eighty four patients were recruited in this study, which comprised 86 patients with CHD, 62 with CHD and FHG, 46 with CHD and T2DM, and 90 patients without the above conditions. The sequences of the PON1 gene L55M polymorphism were analysed by the SNPs methods. RESULTS: The frequency of allele L was 0.86. No significant differences in the frequencies of the three genotypes (LL, LM, and MM) were found among the four groups of patients. The L allele of PON1 L55M was not associated with the number and range of coronary arterial lesions (chi2 = 0.56, P = 0.76; chi2 = 0.11, P = 0.74, respectively). CONCLUSION: PON1 gene L55M polymorphism exists in Han populations in Guangdong, with LL as the main genotype. PON1 L55M gene polymorphism and allelic genes are not associated with the level of fasting blood glucose or diabetes mellitus. There is no evidence to support the relationship between PON1 L55M gene polymorphism and coronary artery lesions. PMID- 19292049 TI - [Association of interleukin-1B gene polymorphisms with coronary heart disease]. AB - OBJECTIVE: To determine whether polymorphisms in interleukin-1B gene promoter -31 and exon 5 +3953 loci are associated with coronary heart diseases (CHD) in Chengdu Han population. METHODS: Two SNPs of IL-1B gene (+3953C/T and -31T/C) in 100 patients with CHD (CHD group) and 144 healthy controls in Chengdu were analysed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Two genotypes at IL-1B + 3953 locus (CC and CT) and 3 genotypes at IL-1B -31 locus (CC, TC and TT) were identified. The -31T alleles carriers were associated with a significantly increased risk of CHD as compared with the non-carriers (OR = 2.12, 95% CI: 1.45-3.09, P < 0.01). Genotypes and allele frequencies at IL-1B + 3953 locus in CHD cases did not differ from the controls. CONCLUSION: IL-1B -31T/C polymorphism may contribute to the risk of developing CHD in Chengdu Han population. PMID- 19292050 TI - [Relationship among TNF-alpha gene promoter -308 site polymorphism, the levels of maternal serum TNF-alpha, and the mRNA expression placental TNF-alpha in preterm labor]. AB - OBJECTIVE: To investigate the role of tumor necrosis factor-alpha (TNF-alpha) in preterm labor and gene polymorphisms of TNF-alpha in genetic susceptibility to preterm labor. METHODS: We collected maternal serum and partial placental tissues from 46 cases of preterm labor(21 cases of infectious proup with chorioamnionitis, 25 cases of noninfectious group without chorioamnionitis), while obtained maternal serum of 50 cases and placental tissues of 20 cases of term labor in contemporary period. TNF-alpha mRNA in placental tissue was tested by RT-PCR, Maternal serum level of TNF-alpha was measured by ELISA. Based on the genotype results of preliminary studies on TNF-alpha promoter -308 site, we analyzed the difference of placental TNF-alpha mRNA, maternal serum levels of TNF alpha in different genotypes, and the relationship of different genotypes to chorioamnionitis. RESULTS: The levels of maternal serum TNF-alpha and the mRNA expression of placental TNF-alpha in preterm labor with chorioamnionitis were significantly higher than those of preterm labor without chorioamnionitis and term labor (P < 0.05). There was no significant difference between those of term labor and preterm labor without chorioamnionitis (P > 0.05). In patients with preterm labor, the levels of placental TNF-alpha mRNA and serum TNF-alpha were statistically significant higher in GA(AA) than in GG(P < 0.05), and GA(AA) has significant relationship with chorioamnionitis Codds ratio (OR) 4.22, 95% CI 1.197, 14.8963. CONCLUSION: It suggested that TNF-alpha -308 polymorphism was associated with the genetic susceptibility to preterm labor. TNF-alpha and its gene polymorphism can be a useful marker for early prediction and diagnosis of preterm labor and intrauterine infection, and may be a genomic index of poor prognosis. PMID- 19292051 TI - [Histological study of miniscrew implanted in lower mandibles of juvenile and adult Beagle dogs]. AB - OBJECTIVE: To examine the histological discrepancies between juvenile and adult Beagle dogs at different concrescence times after miniscrew implants. METHODS: Miniscrew implants were performed in six juvenile Beagle dogs and six adult Beagle dogs. The space between the fourth premolar root and first molar root, and the spaces in distal and mesial of M1 root were picked up for the implants of the 48 miniscrews. The lower jaw specimens including the miniscrews were harvested 3 and 12 weeks after the implants for the histological examinations and bone implant contact (BIC) calculations. RESULTS: There was no miniscrew falling off or becoming loose. The miniscrews had favorable biological consistencies with the tissues around them. The osteoblasts and osteoclasts showed active functions in the peri-bones of the minisrews. The BIC became higher when the healing time was prolonged. The juvenile Beagle dogs had lower BIC than the adults 3 weeks after the implants. But the BIC of the juvenile dogs surpassed the adults 12 weeks after the implants. CONCLUSION: The osseointegration of the miniscrews is hindered by the poorer bone quality of the juvenile Beagle dogs. But the early osseointegration deficiency can be made up by the rapid development and growth of the bones of the juvenile dogs. PMID- 19292052 TI - [Effects of cardiotrophin-1 on differentiation of cardiomyocyte-like cells induced from rat bone marrow mesenchymal stem cells]. AB - OBJECTIVE: To investigate the effects of cardiotrophin-1 (CT-1) on differentiation of induced rat bone marrow mesenchymal stem cells (BMMSCs) in vitro with 5-azacytidine (5-aza) for the purpose of elucidation of the cellular biological mechanisms. METHODS: BMMSCs isolated from femur of rats were divided into four groups: untreated group as control (group A); 0.1 nmol/L CT-1 added to medium (Group B); induced with 10 micromol/L 5-aza only (Group C); induced with 10 micromol/L 5-aza combined with 0.1 nmol/L CT-1 added to medium (Group D). After 4 weeks of induced culturing, the differentiation of induced myocyte like cells were estimated, levels of cardiac troponin-T (cTnT) by immunohistochemical staining and ultrastructure of induce-cultured BMMSCs were determined and mRNA expression of alpha-actin, beta-myosin heavy chain (beta-MHC), Nkx2.5, GATA4 were analyzed by real time polymerase chain reaction (RT-PCR). RESULTS: After 4 weeks of induced culturing, morphological characteristics of myocyte like cells with the expression of cTnT were observed in group C and D cells. Higher level expression of GATA4, Nkx2.5, alpha- actin and beta-MHC mRNA in group D was observed by comparing with those of group C and the differentiated BMMSCs with formations of myofilaments distinctly were also existed in 5-aza combined with CT 1 treatment group. CONCLUSION: This study suggests that induced culturing of BMMSCs in the presence of 5-aza combined with CT-1 can enhance cardiomyocytic characteristics. CT-1 upregulates the expression of GATA4, Nkx2.5, alpha-actin and beta-MHC mRNA, and rapidly promotes the differentiation and maturation of cardiomyocyte-like cells differentiated from BMMSCs induced with 5-aza. PMID- 19292053 TI - [Effect of a traditional Chinese medicine compound on chronic heart failure in guinea-pigs]. AB - OBJECTIVE: To investigate the effect of a traditional Chinese medicine compound (CTMC) on chronic heart failure (CHF) in guinea-pigs. METHODS: The CHF of guinea pigs were induced by repeated injection of hyodemic isoproterenol. The hemodynamics, organ (heart, lung, liver and kidney)/body weight ratio, pathological changes, and serum cTn-I and CK-MB were measured to determine the effectiveness of the traditional Chinese medicine treatement. RESULTS: The LVDP and LVEDP were decreased and the absolute value of + dp/dt(max) and - dp/ dt(max) were increased by the administration of 10 mg/kg, 20 mg/kg and 30 mg/kg of the compound tablets. The effect increased with doses. The traditional Chinese medicine also decreased the area of myocardial necrosis and the degree of injury to myocardiacyte. The intervention group had lower serum cTn-I and CK-MB levels than the controls. CONCLUSION: The compound tablets can improve the left ventricular diastolic function of CHF and reduce the myocardial damage in a dose dependent manner. PMID- 19292054 TI - [Effects of neregulin on rhesus monkey heart failure induced by rapid pacing]. AB - OBJECTIVE: To investigate the effects of Recombined Human Neuregulin (NRG) on the expression of Glucose transporter (Glut1) in myocardium tissue of Rhesus Monkeys with Pacing-induced Heart Failure and the heart function. METHODS: Twenty four rhesus monkeys were randomly divided into three groups (shame operated group, heart failure group and NRG treated group), each with 8 monkeys. Heart failures were induced by rapid pacing (240 heart beats/min). Daily intravenous injection of recombined human NRG [3 microg/(kg x d)] and normal saline were given to the monkeys for 10 days for the NRG treated group and heart failure group respectively. Hemodynamic measurements including +dp/dt(max) and left ventricular systolic, end-diastolic blood pressures (LVSP and LVEDP) were conducted. The RT PCR was applied to detect the expression level of PKB, Glut1 mRNA in the left ventricular cardiac muscle. RESULTS: The monkeys in the heart failure group had lower levels of + dp/dt(max) and LVSP and higher levels of LVEDP than those in the shame operated group (P < 0.05). The monkeys in the NRG treated group had higher levels of +dp/dt(max) than those in the heart failure group (P < 0.05). Lower expression of PKB, Glut1 mRNA in the heart failure group was observed compared with the shame operated group (P < 0.05) while the NRG treated group had a higher expression level of PKB,Glut1 mRNA by compared with the heart failure group (P < 0.05). CONCLUSION: Recombined human NRG can relieve heart failure syndroms through up-regulating the expression of PKB, Glut1 mRNA and improving energy supply of the ischemic cardiac muscle. PMID- 19292055 TI - [Study on the expression of activin-A in myocardial ischemia postconditioning]. AB - OBJECTIVE: To analyze the relationship between the hydroxy free radical (* OH) and the activity of lactate dehydrogenase (LDH) and to study the potential role of activin-A in myocardial ischemia postconditioning for the protection of myocardium. METHODS: Forty healthy adult Sprague-Dawley rats (gender unconcerned) were randomly divided into four different groups (n = 10 in each group): sham operation, ischemia (I), ischemia/reperfusion (I/R) and ischemia postconditioning (postcon). In anesthetized open-chest rats, the left anterior descending artery (LAD) was ligated. Sham operation: the surgical procedure was identical to other groups, but the LAD was ligated for only 30 min, and not occluded. Ischemia: the LAD was occluded for 30 min; Ischemia/reperfusion: the LAD was occluded for 30 min and reperfused for 1 h; Postconditioning: after LAD occlusion for 30 min, at the start of reperfusion, three cycles of 30 s reperfusion and 30 s LAD re occlusion preceded the 1h of reperfusion. The expression of activin-A in myocardium was detected by ELISA in all groups. The ability of suppression on the hydroxy free radical (* OH) and the activity of lactate dehydrogenase (LDH) in myocardium taken from each group were measured by spectrophotometer. RESULTS: Compared with the sham-operated group, the expression of activin-A was significantly elevated in ischemia, ischemia/reperfusion and postconditiong groups, with the highest in the ischemia/reperfusion group. And postconditioning significantly decreased the expression of activin-A in the ischemia/ reperfusion injured myocardium [(343.05 +/- 25.8) microg/mg protein vs (470.6 +/- 33.1) microg/mg protein, P < 0.05]. The expression of activin-A was negatively correlated with the ability of suppression on the hydroxy free radical and LDH separately. CONCLUSION: The result indicated that activin-A was involved in myocardial ischemia/reperfusion injury, and down-regulation of activin-A might be one of the cardioprotective mechanisms of postconditioning. PMID- 19292056 TI - [Effect of Astragalus mongholicus on expression of hepatocyte growth factor in SD rats with unilateral ureteral obstruction]. AB - OBJECTIVE: To study the effect of Astragalus mongholicus (AM) on the expression of hepatocyte growth factor (HGF) in SD rats with unilateral ureteral obstruction (UUO) and to elucidate the mechanisms underlying the renoprotective effects of AM. METHODS: Fifty four Sprague-Dawley rats were randomly divided into 3 groups: sham-operation group (SOR), UUO group (UUO) and UUO + AM group (AM). After administration of AM[10 g/ (kg x d)] for 3, 7 and 14 days,the histological changes of renal interstitial tissues were observed dynamically, and renal damage including tubular impairment and interstitial fibrosis were quantified on HE and Masson stained tissue sections. The protein expression of HGF and alpha-smooth muscle actin (alpha-SMA) was measured by immunohistochemistry. The mRNA expression of HGF and alpha-SMA were determined by real-time PCR. The expression of HGF and its receptor (C-met) were assessed by Western blot. RESULTS: Renal damage was exacerbated and the expression of alpha-SMA was significantly increased in UUO group compared with those of SOR group (P < 0.05) at each time point. HGF and C-met expression peaked at the 7th day after UUO and then decreased greatly. After AM intervention, tubular impairment and interstitial fibrosis were alleviated, up-regulations of alpha-SMA expressions was significantly suppressed, whileas the expression of HGF and C-met were significantly increased when compared with UUO group (P < 0. 05) at each time point. CONCLUSION: AM can ameliorate renal interstitial fibrosis induced by UUO in rats. The mechanisms of its antifibrotic effects may be related with the up regulation of HGF and C-met expression, and the suppression of tubulo-epithelial mesenchymal transdifferentiation in renal intersitial progress. PMID- 19292057 TI - [Effects of ginsenoside Rb1 on TGF-beta1 induced p47phox expression and extracellular matrix accumulation in rat renal tubular epethelial cells]. AB - OBJECTIVE: To investigate the effects of Ginsenoside Rb1 (G-Rb1) on the oxidative damage and extracellular matrix accumulation in rat renal tubular epethelial cells induced by transforming growth factor-beta1 (TGF-beta1). METHODS: Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, 10 ng/mL TGF-beta1-induced group, G-Rb1 treated groups in which rat renal tubular epethelial cells were treated with different concentration of G-Rb1 (10 ng/mL, 20 ng/mL, 40 ng/mL) after TGF-beta1 induction, G-Rb1 40 ng/mL group and 100 nmol/L DPI(diphenyleneiodonium, an inhibitor of NADPH oxidase) group. Intracellular reactive oxidative species (ROS) level was measured by flowcytometry. p47phox protein expression was assessed by immunohistochemistry and western blotting method. The expressions of collagen I (Col-I) and fibronectin(FN) gene were measured by real-time PCR analysis. The protein level of Col-I and FN were quantitatively detected by enzyme-linked immunosorbent assay. RESULTS: TGF-beta1 at 10 ng/mL significantly increased the intercellular ROS production and p47phox expression (P < 0.05). The levels of Col-I and FN were also significantly up-regulated with the stimulation of 10 ng/mL TGF-beta1 (P < 0.05). Compared to TGF-beta1-induced group, G-Rb1 and DPI depressed TGF-beta1 induced ROS production and p47phox overexpression. Meanhile, G-Rb1 and DPI decreased the levels of Col-I and FN. CONCLUSION: G-Rb1 could inhibit TGF-beta1 induced ROS production and decrease the levels of Col-I and FN in a dose dependent manner. The mechanism might be partly related to the suppression of p47phox expression. PMID- 19292058 TI - [Effect of tanshinone II A on NB4 cell induced procoagulant activity in human umbilical vein endothelial cells]. AB - OBJECTIVE: To investigate the effect of tanshinone II A on the procoagulant activity (PCA) of human umbilical vein endothelial cells (HUVEC) induced by acute promyelocytic leukemia (APL) cell line NB4 cells. METHODS: The HUVEC were incubated for 6, 12, and 24 hours in different tanshinone II A conditioned medias (Tan II A-NB4-24h-CM, Tan II A-NB4-72h-CM, Tan II A-NB4-120h-CM). Then the HUVEC were incubated for 6, 12, 24, and 72 hours with Tan II A-NB4-120h-CM and different concentrations of Tan II A (0, 0.25, 0.5, 1.0 microg/mL). The HUVEC lysates were obtained by three repeated freezing and thrawing. Their PCA were tested using the one stage clotting assay. The activity of tissue factor (TF : act) was tested using the chromogenic substrate assay. The control groups included 0.3 microg/mL ATRA, 0.01% DMSO and RPMI 1640. RESULTS: Tan II A-(72 h,120 h)-NB4-CM elevated PCA of HUVEC and six hours of incubation in the 120 h NB4-CM had the greatest PCA. The PCA of HUVEC in the 1.0 microg/mL Tan II A-NB4 CM was the same as in the 0.3 microg/mL ATRA-NB4-CM. (2) The NB4-CM induced PCA of HUVEC decreased with 5.0 microg/mL of Tan II A, at a level similar to the decrease with 0.3 microg/mL of ATRA. Less than 5.0 microg/mL of Tan II did not reduce the NB4-CM induced PCA of HUVEC. (3) Both Tan II A 120 h-NB4-CM and ATRA 120 h-NB4-CM elevated the TF : act of HUVEC. The TF : act reached the peak after 6 hours of incubation. The Tan II A 120 h-NB4-CM maintained the peak level of TF : act at the 12th hour and fell to the base line at the 24th hour. The ATRA 120 h NB4-CM induced TF:act dropped down with time after reaching its peak at the 6th hour. (4) The 1.0 microg/mL of Tan II A did not reduce the TF : act of HUVEC induced by the Tan II A 120 h-NB4-CM. But the 0.3 microg/mL of ATRA reduced the TF : act of HUVEC at the 6th hour. CONCLUSION: TanIIA-NB4-CM increases PCA and TF : Act of HUVEC. TanIIA decreases PCA of HUVECs induces by TanIIA-NB4-CM. PMID- 19292059 TI - [Influence of dexamethasone on expression of 11beta-HSD2 in primary cultured cytotrophoblasts from human preterm placenta]. AB - OBJECTIVE: To study the influence of dexamethasone (DEX) on the expression of 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) in primary cultured cytotrophoblasts from human preterm placenta. METHODS: Placenta villous cytotrophoblasts from preterm birth were cultured with the protocol of enzymatic dissociation and tissue incubation method. After isolation and identification, cytotrophoblasts were treated with the repeated administration of DEX (100 nmol/L), or DEX-RU486 (1 micromol/L) for 7 days, in which DEX was not added into the culture at 4th day. Cytotrophoblasts were collected everyday, and the expression levels of 11beta-HSD2 mRNA and protein were determined by real-time fluorescence quantitative PCR and Western blot method. RESULTS: After the treatment of DEX (100 nmol/L), the expression of 11beta-HSD2 mRNA and protein in cytotrophoblast increased in the first three days (P < 0.05). At 4th day, 11beta HSD2 mRNA and protein declined in the absence of DEX. In 5th-7th day, the increase of 11beta-HSD2 expression were resumed when cytotrophoblasts received DEX again (P < 0.05). With the treatment of DEX and RU486 (l micromol/L), both mRNA and protein level of 11beta-HSD2 in cytotrophoblasts were lower than those with DEX alone, but there was not significantly different (P > 0.05). CONCLUSION: Repeated administration of DEX can upregulate the expression level of 11beta-HSD2 in primary cultured cytotrophoblasts from preterm placenta. Cytotrophoblasts from preterm birth may have the ability to protect the infants by the mechanism of 11beta-HSD2 regulation. PMID- 19292060 TI - [Bone targeting of antitumor conjugate phenamine acid aryolysine-hexanedioic acid bridge grafting-bisphosphonates]. AB - OBJECTIVE: To develop a new bone targeting antitumor therapy system which uses diphosphonate and bone-seeking nuclide to enhance the coordinated effects of chemotherapy and radiotherapy on bones, and to validate the targeting of the new therapy system in vitro and in vivo. METHODS: Phenamine acid caryolysine was connected to bisphosphonates, and then combined with radioactive nuclide 153Sm to establish a new bone targeting chemotherapeutic and radioactive drug for bone cancers. The targeting of this new therapy system was validated by hydroxyapatite crystal absorbing test and body distribution in vivo methods. RESULTS: The optical spectrum of the phenamine acid caryolysine-bisphosphonates conjugate detected by the nuclear magnetic resonance was consistent with the standard structure of synthesized drugs. The conjugate had good absorbability to hydroxyapatite crystals. The body distribution of the conjugate showed higher radiocounting in bones than in other tissues. CONCLUSION: The conjugate has a structure that is consistent with the active compound targeting bone tumors. PMID- 19292061 TI - [The expression of APE1 and its correlation with prognostic significance after 252Cf radiotherapy in cervical cancer]. AB - OBJECTIVE: To investigate the expression feature of the apurinic/apyrimidinic endonuclease (APE1) and its correlation with clinicopathology and prognostic significance after 252Cf radiotherapy in cervical cancer. METHODS: The expression of APE1 was detected by immunohistochemistry technique in 89 cases of cervical cancer (treated by 252Cf), 15 cases cervical intraepithelial neoplasia (CIN) and 10 cases of normal cervical tissue, and its association with clinicopathological data as well as prognosis were analyzed. RESULTS: The expression of APE1 in cervical cancer is higher significantly than that in normal cervical tissue and CIN (P < 0.01). In normal cervical tissue and CIN, the APE1 express was located in the nucleus. In cervical cancer, the APE1 express was located in the nucleus (59), cytoplasm (8) or nucleus and cytoplasm (22), the location of APE1 was related with FIGO stage and pathological grade (P < 0.01), and not related with lymph node metastasis. The level of APE1 express related with FIGO stage, pathological grade and lymph node metastasis (P < 0.05), and not related with age and pathological type. The Kaplan-Meier survival analysis demonstrated that the survival time of the group of APE1 nucleus expression (median survival time is 70.9 months) and the group of APE1 low expression (median survival time is 75.8 months) is longer significantly than that of the group of APE1 cytoplasm expression (median survival time is 57.8 months) and the group of APE1 high expression (median survival time is 56.5 months) (P = 0.025, 0.001). CONCLUSION: The dystopic express of APE1 might play a pivotal role in carcinogenesis and progression of cervical cancer, and the express of APE1 might estimate the prognosis after 252Cf radiotherapy. PMID- 19292062 TI - [A clinical trial for homoharringtonine and low-dose cytosine arabinoside combined with G-CSF or GM-CSF to treat the relapsed or refractory acute myeloid leukemia (AML), geriatric AML and advanced myelodysplastic syndromes]. AB - OBJECTIVE: To evaluate the effectiveness and toxicity of the regimen combined homoharringtonine, low-dose cytarabine with G-CSF or GM-CSF (HAG regimen) in treating patients with relapsed or refractory AML, geriatric AML and advanced myelodysplastic syndromes (MDS). METHODS: Forty patients with AML or advanced MDS were treated with HAG regimen for remission induction and consolidation therapy. All of them were followed up till April 2006. Results 20 of them (50%) achieved complete remission (CR), including 46.2% patients with relapsed or refractory AML, 60% elderly patients with primary AML who were either untreated or treated with only one course of induction therapy previously, and 66.7% patients with MDS RAEB. After a follow-up of 6- 47 (median 23) months from the date of remission, the median times of relapse-free survival and overall survival were (7.0 +/- 1.1) and (28 +/- 12.3) months, respectively. Myelosuppression was the most significant toxicity. The incidences of infection and hemorrhage which exceed grade II were 43.8% and 37.5%, respectively. Non-hematologic adverse effects were minimal. CONCLUSION: The HAG regimen presented effective and well-tolerated. It seems promising for the treatment of relapsed or refractory AML, geriatric AML and advanced MDS. PMID- 19292063 TI - [Valproic acid induced intracellular GSH-redox imbalance and apoptosis of leukemic cells resistant to dexamethasone and doxorubicin]. AB - OBJECTIVE: To investigate the anti-neoplastic effects of Valproic acid (VPA) on leukemic cells, especially drug-resistant lines, and to investigate whether modulation of GSH-redox status is involved in VPA-induced apoptosis. METHODS: After the treatment of VPA at various concentrations for indicated times, cellular proliferation of the Jurkat, CEM, HL-60, K562, K562/AO2 cells were evaluated via MTT assay; and the activities of Caspase-3, Caspase-8 and Caspase-9 were quantitatively analyzed by colorimetric assay. The morphological change and cell cycle distribution were also examined on Jurkat (Dexamethasone-resistant) and K562/AO2 (Doxorubicin-resistant) cell lines. The levels of intracellular glutathione/glutathione disulfide (GSH/GSSG) and the activities of the typical antioxidant enzymes, i.e., glutathione reductase (GSH-Rd) and glutathione peroxidase (GSH-Px), were measured on cell lysates of Jurkat and K562/AO2 cell lines prior to and after VPA treatment. Apoptosis rates of Jurkat and K562/AO2 cells treated with VPA along or in combination with N-acety-l-cysteine (NAC), catalase (CAT) or DL-buthionine-(S,R)-sulfoximine (BSO) were determined by Annexin V/propidium iodide (PI) staining with flow cytometry analysis. RESULTS: At concentrations comparable with that achieved at clinical settings, VPA inhibited cell proliferation, activated Caspase-3, 8, and 9, and induced cell cycle arrest in Jurkat and K562/AO2. A rapid decrease in GSH-Rd and GSH-Px activities and GSH content in Jurkat and K562/AO2 were detected after VPA treatment. Co-administration of NAC or CAT attenuated VPA-induced apoptosis. CONCLUSION: VPA inhibit cell proliferation, induce cell cycle arrest and apoptosis in drug-resistant leukemic cells. Apoptosis correlates with down regulation of intracellular GSH and disruption of intracellular GSH-redox balance, possibly through inhibition of glutathione reductase and glutathione peroxidase. PMID- 19292064 TI - [Isoniazid prophylaxis for pulmonary tuberculosis in Chinese patients with rheumatoid arthritis receiving long-term methotrexate therapy]. AB - OBJECTIVE: To investigate the efficacy and safety of Isoniazid prophylaxis (INHP) for tuberculosis in Chinese patients with rheumatoid arthritis (RA) who receive long-term Methotrexate (MTX) therapy. METHODS: Two hundred and one (201) patients with RA were randomized to initial treatment with either MTX/Isoniazid (INH) [MTX, 0.1-0.3 mg/(kg x week), maximal 20 mg/week; INH, 5 mg/(kg x d), max 300 mg/d] or MTX alone [0.1-0.3 mg/(kg x week), maximal 20 mg/week]. The doses of MTX remained the same after 6-month INH treatment. All patients were followed up for 36 months, the incidence of pulmonary tuberculosis (TB) was investigated, while the toxicity of INH and MTX were assessed. RESULTS: There were 77 patients completed INHP. Two withdrew due to unwilling to participate. Treatment was discontinued in 4 cases (4.9%) owing to toxicity: increasing ALT/AST in two (2.5%), decrease of white blood cell (WBC) in one (1.2%) and anaphylaxis in one (1.2%). In control group, 5 patients withdrew for personal reasons. The incidence of adverse effect due to MTX in this group was 6.2%: Hepatotoxicity appeared in 4 patients (3.5%) and decrease of WBC was seen in 3 (2.7%). Nine patients (8.5%) developed TB in control group, and 1 patient (1.3%) developed TB 14 months after INH treatment. The risk of TB infection in control group was 6 times more than that in INHP group (chi2 = 4.47, P < 0.05). CONCLUSION: INHP is safe and effective to prevent TB in RA patients treated with MTX. PMID- 19292065 TI - [Effect of carbon dioxide pneumoperitoneum on the translocation of intestinal endotoxin/bacteria in rats]. AB - OBJECTIVE: To evaluate the effect of carbon dioxide pneumoperitoneum on the translocation of intestinal endotoxin/bacteria. METHODS: The ampicillin-resistant E. coli JM109 labeled with plasmid PUC18 was given by gavage to 48 rats to detect the translocation of intestinal bacteria. Hydrochloric acid was peritoneal inoculated to the rats to induce acute chemical peritonitis. The rats were randomly allocated to the control group without pneumoperitoneum (n = 16), the experimental groups with low insufflation (5 mmHg of carbon dioxide, n = 16) and with high insufflation (15 mmHg of carbon dioxide, n = 16). The blood endotoxin in the femoral veins were detected every hour. The ampicillin-resistant E. coli JM109 were identified by restricted digest and agrose gel-electrophoresis. The rats were killed at the end of the 3 hour experiment and blood samples from the portal and mesenteric lymph nodes were taken for culture. The levels of SIgA in the plasma and the intestinal mucosal were detected by radioimmunoassay. RESULTS: The blood endotoxin in the femoral veins of the rats increased significantly 1 hour after the induction of pneumoperitoneum with 15mmHg of carbon dioxide (P < 0.01) and continued to increase with time. The plasma and intestinal mucosal SIgA of the rats decreased significantly (P < 0.01) after 3 hours of exposure to 15 mmHg of carbon dioxide pneumoperitoneum. The bacterial translocation rates from intestinal to portal were 0%, 25% and 50% for the control, 5 mmHg and 15 mmHg groups respectively, and 12.5%, 25%, and 75% from intestinal to mesenteric lymph nodes for the three groups respectively. CONCLUSION: Carbon dioxide pneumoperitoneum increase intestinal Endotoxin/bacterial translocations and the effect increases with pressures. PMID- 19292066 TI - [Effect of weight on bone mineral density in ovariectomic rats]. AB - OBJECTIVE: To evaluate the effect of weight on bone mineral density in ovariectomic rats. METHODS: Forty 6-month-old Wistar female rats were randomly divided into the sham surgical group and the ovariectomic group. The weights of the rats and the bone mineral densities and bone mineral contents of the femurs of the rats were measured eight weeks after the operations. RESULTS: The body weights of the ovariectomic rats increased significantly. The vaginal cytology of the ovariectomic rats showed no estrous cycle. The areas of the uterus transverse section of the ovariectomic rats deceased significantly. The depth of endometrium and its epithelia, and the area percentage of endometric gland of the ovariectomic rats were smaller than those of the sham surgical controls. There were no statistically differences in the BMD and BMC between the ovariectomic rats and the sham surgical controls. However, the BMC/weight ratio of the ovariectomic rats decreased significantly. The linear stepwise regression analysis indicated a positive correlation between BMC and body weight and a negative correlation between BMC and the weight differences pre- and post operations. The BMC/weight ratio also had a negative correlation with the weight differences. CONCLUSION: The osteoporotic animal model after ovariectomy has been successfully established. The certain increased body weight after ovariectomy may slow the lose of bone mass. But the over increasing in body mass lead to BMC/weight decling, caused by osteoporosis. PMID- 19292067 TI - [Pharmacokinetics of PEG recombinant human interleukin-6 in rats]. AB - OBJECTIVE: To study the pharmacokinetics and tissue distribution of PEG-rhIL-6 in rats after a single dose administration. METHODS: Pharmacokinetics and distribution of PEG-rhIL-6 in rats were studied by 125I isotope tracing method. Pharmacokinetic analysis was performed using 3P97 computer software. RESULTS: PEG rhIL-6 declined in one-compartment model with half-lives of 10.44-11.37 h for t1/2 Ka, 19.77-21.53 h for t1/2 Ke and 20.51-21.96 h for T(pcak), respectively. PEG-rhIL-6 was mainly distributed in blood and excreted via urine. CONCLUSION: The half-lives of PEG-rhIL-6 are prolonged after being modified by PEG. PMID- 19292068 TI - [Isolation, purification and primary culture of rat pancreatic beta-cells]. AB - OBJECTIVE: To isolate and purify rat pancreatic beta-cells and to explore the best conditions for the primary culture of the pancreatic beta-cells in vitro. METHODS: The pancreas of Norman Wistar rats were digested by collagenase V. The islets were purified by mesh sieve. The activity of the islets was stimulated by different concentrations of glucose and detected by dithizone dye. The purified islets were put into RPMI-1640 nutritive medium for culture overnight. The cultured islets were digested again with trypsin and DNAase to obtain the suspension containing single pancreatic cells. The beta-cells were separated and purified in a fluorescence-activated cell sorter (FACS) in the medium containing 2.8 mmol/L glucose. The purified beta-cells were identified by immunohistochemistry and glucose stimulating test. Ham's F-10 with different concentrations of glucose and 3-Isobutyl-1-methylxanthine (IBMX) were used as nutritive medium for the primary cell culture for 24 hours. The best conditions for the culture were identified. RESULTS: An average of 550 +/- 90 islets with fine activities were obtained per rat. The purification with FACS obtained about 5688 beta-cells per rat, with a recovery rate of (93.69 +/- 1.26)% and a purity of (85.5 +/- 1.24)%. A concentration of 10.0 mmol/L and 16.0 mmol/L glucose in primary culture for 24 hours produced the highest survival rates of beta-cells, but IBMX did not increase the survival rates of beta-cells. CONCLUSION: FACS is effective in purifying pancreatic beta-cells from the suspension with a medium containing 2.8 mmol/L glucose. Pancreatic beta-cells maintain relatively high activities in Ham's F-10 medium containing 10.0-16.0 mmol/L glucose in primary culture. PMID- 19292069 TI - [Application of laser capture microdissection and surface-enhanced laser desorption ionization time-of-flight mass spectrometry to screen biomarkers for early diagnosis of lung adenocarcinoma]. AB - OBJECTIVE: To improve diagnostic methods to screen biomarkers for early diagnosis in lung adenocarcinoma by employing laser capture microdissection (LCM) and surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and support vector machine (SVM). METHODS: Frozen sections of thickness 8 microm were made using 6 cases of fresh lung cancer tissues and 4 cases of matched normal lung tissues. The sections were stained by improved HE solution. The homogeneous adenocarcinoma cells and normal cells were collected by LCM in each sample, and then SELDI profiles based on PBS II+SELDI-TOF-MS (IMAC protein chip) were analyzed using SVM. RESULTS: High quality cell samples were obtained by LCM quickly and precisely from normal specimens and diseased tissues without interstitium, inflammation and necrosis. Eighty four differential protein peaks were found. Top ten of them were identified as candidate biomarkers; six proteins were significantly weakly expressed in lung cancer tissue compared to normal tissues, but the other four protein were over-expressed (P < 0.05). Every candidate biomarker has undergone the blind-cross-test. Each of them can separate the lung cancer from normal samples with a sensitivity of 100% and a specificity of 100%. The 3191 m/z was considered as disease marker of lung adenocarcinoma. CONCLUSION: The method combined LCM with SELDI-TOF-MS may be able to screen potential biomarkers to distinguish lung cancer from healthy tissue with high sensitivity and specificity, which could improve early diagnosis for lung cancer. PMID- 19292070 TI - [Study on the detection method for determining the bioactivity of recombinant human interleukin-22]. AB - OBJECTIVE: To develop a method for determining the bioactivity of recombinant human IL-22. METHODS: pSTAT3-TA-Luc and pcDNA3. 1 plasmids were co-transfect to HepG2 cells to generate stable HepG2/ STAT3 cell line with G418 screen. After treating cell with IL-22, the luciferase activity was assayed. Orthogonal test was used to optimize the assay condition, and then the reproducibility and specificity of the assay was checked. RESULTS: The best condition for this assay are: cell density as 4 x 10(5)/mL, stimulating time as 4 hours, luciferase substrate as 50 microL. 50% effective dose (ED50) of IL-22 assayed by this method is 16.89 ng/mL, relative standard deviation (RSD) is 7.09%. Neutralizing antibody test shows the high specificity. Comparing with ELISA, the method described here has more advantages, including higher stability, easier performance and less cost. CONCLUSION: Luciferase reporter gene assay method is a fast, sensitive, reproducible method for IL-22 bioactivity determination. PMID- 19292071 TI - [New directions in medical thinking?]. PMID- 19292072 TI - [New trends in the classification of personality disorders]. AB - The author makes an overview of the latest theoretical developments and of the critical views on the current classification of personality disorders. The major topics of the ongoing debate over the typology of personality disorders as presented by DSM-IV-TR are listed as the difficulties that remain in clarifying the clinical criteria, despite the clinical assessment tools and the self assessment questionnaires that are usually employed in research. The solution to the dilemma categorical vs. dimensional in personality disorders taxonomy, as suggested by some current authors in dimensionally quantifying the diagnosis criteria for the classic types of personality disorders has received much attention. But the relevant practical solutions need more longitudinal future studies done on a congruent agenda. PMID- 19292073 TI - [The gastro-esophageal reflux disease and its psycho-somatic component]. AB - Different factors may be involved in the occurrence of gastro-esophageal reflux disease (GERD) symptoms. This review presents the importance of psychic factors in GERD symptom genesis, especially in patients with no esophagitis. In addition, these factors could be involved in the pharmacological therapy, with educational and behavioral implications increasing the cost-efficacy of the therapy. PMID- 19292074 TI - [Radiofrequency ablation of malignant liver tumours]. AB - Both primary and secondary liver tumors are common problems. Hepatocellular carcinoma (HCC) is a tumor with rapid progression and poor prognosis. The standard in the treatment of liver tumors is surgical resection. The majority of patients with primary or metastatic hepatic tumors are not candidates for resection because of tumor size, location near major intrahepatic blood vessels precluding a margin-negative resection, multifocality, or inadequate hepatic function due to coexistent cirrhosis. Only 20-30% of patients undergoing potentially curative liver resection will not suffer from tumor recurrence. Radiofrequency ablation (ARF) is an evolving technology used to treat patients with non-removable primary and metastatic hepatic cancers. ARF produces coagulative necrosis of tumor through local tissue heating. Liver tumors are treated percutaneously, laparoscopically, or during laparotomy using ultrasonography to identify their position and guide the ARF needle electrode. The indications, treatmentplanning, and limitations of hepatic ARF must be defined. ARF of hepatic malignancies is a safe and promising technique to produce coagulative necrosis of non-removable hepatic malignancies. Experience with this treatment is not yet rich enough to establish long-term outcomes. PMID- 19292075 TI - [Under-evaluated extrahepatic manifestations of cholestasis]. AB - Biliary stasis can occur in many different diseases. Pruritus, metabolic bone disease, deficiencies of fat-soluble vitamins, steatorrhea, hyperlipidemia and fatigue represent the major extra-hepatic manifestations of cholestatic liver disease that considerably affect the patient's quality of life. The present article reviews pathogenetic and clinical aspects of and current therapeutic approaches to extra-hepatic manifestations of cholestatic liver disease. PMID- 19292076 TI - [The implications of nutrition in the obesity of children and adolescents]. AB - Obesity is currently the most prevalent nutritional disease of children and adolescents and a common health problem. Besides the bio-ecologic and socio economic factors (genetical, professional, cultural, educational), the exogenous factors (nutritional) have a pro-obesity action by increasing the caloric intake determining energy imbalance. The authors present the role of nutrition in the occurrence of obesity in children and adolescents and its role in the treatment of the disease. PMID- 19292077 TI - [Five years of kidney transplantation in Iasi]. AB - The activity of Renal Transplant Center Iasi started in November 2000, when we realized the first renal transplant from a live donor. Since then, 46 renal transplants were successfully realized in our center, to patients aged between 13 47 years (medium age = 30 +/- 5), M/F=27/19, 25 (56.8%) of them selected from HD, 17 (39.9%) from CAPD and 4 (8.7%) pre-emptive. Medium age of donors was 49.1 years (30-64), M/F=29/17. The donors were, in 78.3% of cases, first-degree relatives (parents, siblings). In 10 cases (21.7%) the grafts were donated by "emotionally related"donors (in most cases, spouses). An urologist-cardiovascular surgeon team, performed the transplant operations. There were no important complications during operation. We had one major vascular complication (graft artery thrombosis) in a 13 years recipient, successfully resolved after thrombectomy and stenting. Immunosuppressive therapy associated induction with monoclonal antibodies anti-Tac, cyclosporine, MMF and prednisone. Eight patients from 46 (17.39%) presented acute rejection episodes and all responded at corticosteroids. Medium values of serum creatinine were: 1.54 mg% at 1 month, 1.42 mg% at 6 months, 1.44 mg% at 1 year, 1.21 mg% at 2 years, 1.38 mg% at 3 years, 1.4 mg% at 4 years and 1.2 at 5 years. The survival of patients and donors is 100% and the survival of renal graft--97.1% (one case of chronic allograft nephropathy with lost of renal function). We also present the satisfactory evolution of the 51 renal transplanted patients addressed to our center from different other renal transplantation centers in Romania for management follow up. PMID- 19292078 TI - [Assessing the number of TB cases using the capture-recapture method with two data sources]. AB - AIM: A more accurate assessment of the number of TB cases in Iasi County using the capture-recapture method. METHODS: TB patients diagnosed in Iasi County are included in the Registry of Tuberculosis, and are microbiologically examined. The Registry of Tuberculosis in Iasi County contains data about the diagnosis and treatment of TB patients that live in this area (notification). The second list includes all the patients smear positive or culture positive for TB. This list will be obtained from the Microbiology Laboratory, from the Clinic of Pulmonary Diseases and the Municipal Hospital of Pascani. RESULTS: There are 1,134 notifications, 763 laboratory, 642 overlap and thus 1,255 patients in total. The overlap between the data sources is 51%. The capture-recapture leads to an estimate of 1,348 patients (95% CI: 1,320-1,375). This means that the estimated exhaustive of the notifications is (1,134/1,348) = 84.1% (95% CI: 82.5-85.9%) and the exhaustive of the lab is (763/1,348) = 56.6% (95% CI: 55.5-57.8%)). CONCLUSION: The capture recapture assessment using two data sources estimated a that the notifications are about 84% exhaustive, like WHO estimations (80%). PMID- 19292079 TI - [Current trends in the epidemiology of bloodstream infections in patients with hematological malignancies and solid neoplasms]. AB - OBJECTIVES: to characterize the epidemiological and microbiological spectrum of systemic infections in hematological and oncological patients. METHOD: a prospective study of bloodstream infections was conducted between May 2004 December 2005 in two hospitals from Iasi. RESULTS: We investigated a total of 86 suspected episodes of systemic infections that occurred in 76 selected patients, 40 males and 36 females (average age = 26 years old). There were 39 (45.35%) febrile episodes with positive blood culture and 47 (54.65%) with negative blood culture. The most common microorganisms were gram-positive cocci 12 (30.76%), followed by stable cell-wall deficient forms 9 (23.07%), gram-negative bacteria 7 (17.95%), gram-positive bacteria 5 (12.82%), fungi 4 (10.25%), and anaerobes 2 (5%). From the total of 23 cell-wall deficient form strains (58.97%), 14 returned to their typical morphology (6 gram-positive cocci, 5 gram-positive bacilli, 3 gram-negative bacilli) and 9 were L-stable. CONCLUSIONS: 1) The isolation rate of 45.5% was in accordance with literature reports; similar percentages of gram positive and gram-negative isolates were found (the shift from predominating gram negative to a gram-positive etiology was noted). 2) A high proportion of cell wall deficient forms was found, rarely reported in previous studies. 3) Studies reporting local microbiological findings are necessary because they support an initial empiric antibiotic regimens for prophylaxis or therapy more accurately than from other areas. The right treatment protocols remain to be established. PMID- 19292080 TI - [Clinical epidemiological research on the evolution of rubella cases recorded in some collectives of institutionalized children]. AB - Authors present the results of a retroprospective study to assess the number of rubella cases recorded in medical social assistance units for children, in two districts of Moldavia, Romania, between 1993-2003. The age distribution highlighted a predominance of 11-19 age groups in Botosani and 5-14 in Iasi; and sex distribution, a predominance in females, but with no significant difference. The rubella epidemic peak of year 2003 was observed in both districts. The necessity of Immunization National Program spreading for rubella and rubella congenital syndrome prevention is a reality, taking into consideration the high incidence of this disease in institutionalized children also. PMID- 19292081 TI - [Multidisciplinary approach in pyomyositis early diagnosis]. AB - Pyomyositis is a disease characterized by primary bacterial infection of the skeletal muscles of lower limbs and thorax. It is a very rare disease (100 cases in USA in the last 20 years) but with an increasing occurrence especially among patients with AIDS in an advanced stage or in association with other pathologic conditions like diabetes mellitus, cancer, connective tissue diseases or hematological disorders, long usage of corticosteroid drugs. We are hereby showing the case of a 46-year-old female who presented difficult problems of differential diagnosis until final diagnosis was established. Her life was saved due to an exemplary interdisciplinary collaboration and by using an extremely useful imaging method for musculoskeletal pathology--IRM. The IRM protocol: coronal and axial T2 weighted SE, coronal and axial T1 weighted SE, axial and coronal T2 weighted + Fat Sat was comprehensive and conclusive. The imaging diagnosis established the presence and extension of the muscle abscess, leading to an efficient drainage. After the surgery, the repeated IRM confirmed the drainage efficiency. Unlike the French literature which claims that the muscle abscesses are secondary to a primary center, in this case, and in accordance with Anglo-Saxon literature, the muscle abscess was primary, in absence of any other center of infection. PMID- 19292082 TI - [Dextrocardia of post right pneumonectomy]. AB - The negative pressure achieved in the hemi thorax in the case of lung pneumonectomy has consequences on the mediastinum organ statics. This condition results through the other lung expansion. These changes are common, but rare in adult lung pneumonectomy. In the case of right lung pneumonectomy, the mediastinum moves to the operated part and disturbs the function of the vessels and cavities with thin wall or low pressure (vena cava, right atrium). These aspects are not observed in the case of left lung pneumonectomy. Taking this into consideration the authors present the case of a young woman with right lung pneumonectomy, an intervention made in childhood, at ten years old. After sixteen years of evolution a serious mediastinum re-shuffle was discovered, the heart was moved in the right hemi thorax, with a dextrocardia like aspect. This case is interesting because it raises positive and differential diagnosis problems- dextrocardia, associated disorders, serious complications. PMID- 19292083 TI - [Hypersensitivity myocarditis, a surprising diagnosis. Case report]. AB - Hypersensitivity myocarditis is a rare morphoclinical entity that may cause sudden death through arrhythmia and cardiogenic shock. The focus of the paper is the histological evaluation of the cases to identify the death cause of a 50-year old patient, known to have suffered of hypertrophic cardiomyopathy, infective endocarditis and severe mitral regurgitation, case in which hypersensitivity myocarditis precipitated the evolution of the disease. The basis of the study was a complete cardiac evaluation including the morphological diagnosis on necropsy specimens. The paper shows that hypersensitivity myocarditis is a cause of sudden death through acute cardiac failure, and the setting of a precise morphological diagnosis is very important for an early targeted treatment. PMID- 19292084 TI - Cheyne stokes respiration in stroke patients. AB - Cerebral-vascular events represent a major cause of mortality worldwide. Resulting respiratory complications are common and possibly composing a marker of severe neurological disorder. Cheyne-Stokes respiration is a disorder characterised by recurrent central apnoeas during sleep, alternating with a crescendo-decrescendo pattern of tidal volume, thus constituting an additional burden to the patient. In this paper the authors review the physio-pathological background, clinical presence and latest therapy-related data on Cheyne Stokes respiration in stroke patients. PMID- 19292085 TI - [New developments in the diagnosis of benign prostatic hyperplasia]. AB - The first phase in the diagnosis of patients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia is to exclude another disease. This initial evaluation consist of anamnesis, including the quantification of symptoms according to the International Prostatic Symptom Score, physical examination- with digital rectal examination playing a central role, urinalysis, and renal function assessment; the prostate-specific antigen may suggest prostate cancer or may predict the volume of the prostate and the natural evolution of the disease. If, after the initial evaluation, the diagnosis is still uncertain, further investigations are necessary: imagistic exploration of the urinary tract, urodynamic tests or urethro-cystoscopy. The authors review current knowledge. PMID- 19292086 TI - [Ultrasound of cervix uteri transvaginal in preterm birth]. AB - Premature birth is a social problem due to its implications in the natal mortality and morbidity. This is the reason why it is necessary to establish new methods which could identify pregnant women with high risk of premature birth. Besides the measurable biochemical factors such as fetal fibronectin and salivary estriol, I1-6 sonography is a simple method highly efficient and of reasonable financial costs among all the sonographic means of observing the cervix uteri. The transvaginal sonography is the "golden standard", permitting the most faithful assessment of the pursued parameters--the length of the cervix, the aspect of the internal os (funneling), the cervix index. The experience gathered up to now shows that all of these investigations have a predictive value which is more negative than positive, but when used together they could lead to an improved result. PMID- 19292087 TI - [Specific effects upon hearing due to occupational noise]. AB - Every day, millions of employees are exposed to noise at work and at the risks this can entail. While the noise is most obviously in industries such as manufacturing and construction, it can also be an issue in a wide range of other working environments. One in five workers have to raise their voice to be heard for at least half of the time when they are at work. This paper shortly presents the physical aspects of the noise and its dangerous local level effects on the human body. PMID- 19292088 TI - [Pancreatic islets isolation from the brain death donors]. AB - The paper analyzes a number of 5 cases of pancreatic islet cells isolation, purification and culture extracted from the pancreas of brain-dead donors. For the isolation stage the authors used the "two-step" digestion method. The isolated pancreatic islet cells were morphologically and functionally analyzed by assessing their viability and purity. For the in-vivo experiments the authors used animals with Streptozotocin induced diabetes. The data clearly indicated a high efficiency of collection providing an optimal number of transplantable pancreatic islet cells. PMID- 19292089 TI - [Comparative experimental study of the effect of various locally administered pharmacological agents on the appearance of peritoneal adhesions]. AB - The paper evaluates the intensity of peritoneal adhesions induced by talc administration on Wistar rats, alone and in association with different other substances. The comparative study was performed on 40 rats, separated in 5 groups: group A was injected intraperitoneally with talc, group B with talc and streptokinase, group C with talc and heparin, group D with talc and dexamethasone and group E with talc and dextran. The adherence syndrome was evaluated after 3 weeks, scoring 5 parameters: number, density, strength, vascularization, topography and granulomas. The sum of these 5 parameters was defined as the adherence score. The best results in preventing adhesion formation are obtained with the intraperitoneal administration of dextran and streptokinase. PMID- 19292090 TI - [Surgical resection in esophageal-gastric junction adenocarcinoma]. AB - Between January 1999 and November 2005 there have been a number of 44 cases of cancers of the esophageal-gastric junction. Only 24 of them have been surgically removed. The carcinoma of the esophageal-gastric junction was defined on topographical and histological grounds, complemented by radiological, endoscopical and tomographical aspects. The authors analyzed the location of the tumor, compared the imaging with the intra-surgical findings and chose the surgical strategy. The authors widely used the classification proposed by Siewert and Stein, which should be the reference for such tumors. The distal esophagectomy for type I and II tumors should reach much closer, at a reasonable, oncologically sufficient, distance from the upper limit of the tumor; the transhiatal approach is as good as a trans-thoracic one for an accurate mediastinal lymphadenectomy; an esophago-jejunal anastomosis should not be carried out at any costs, reducing the height of the esophageal resection (a risk of positive resection margins), for the length of the jejunal segment to fit the gap. PMID- 19292091 TI - [The results of the Sommerville-Petit procedure in the treatment of the typical congenital hip dislocation]. AB - The studied group from the Pediatric Surgery and Orthopedics Clinic, Iasi, consisted of 87 cases, which means that a total of 118 hips were treated between September 1998-September 2004. In 116 cases, the extension was performed with trans-femoral Kirschner wire, in a period of 3-7 weeks. After 7-14 days of extension, the authors performed adductor and sometimes psoas tenotomy. The stable orthopedic reduction was obtained in 34 hips, while the 15 cases with inadequate results required open reduction. This method, performed from the beginning after a period of extension, was applied to 69 cases. In 13 patients the iterative open reduction was required. None of the cases associated femoral diaphysis shortening resection. PMID- 19292092 TI - [Diabetic eye in children]. AB - Diabetes mellitus is a new morbidity in children and adolescents. For pediatric patients, it announces the earlier onset of cardiovascular disease, retinopathy, nephropathy, and neuropathy, with the risk of impaired quality of life and premature death. The emergence of diabetes mellitus in young people is believed to be associated with changes in the level of physical activity and nutrition that are ubiquitous in modern society. The authors present a prospective study of 26 children with diabetes mellitus. The average age was 9.1 +/- 2.8 for boys and 12.6 +/- 3.7 for girls. The ophthalmologic disorders founded were: lens modifications in 7.7%, refractive errors in 34.6% and diabetic retinopathy in 11.5%. The study estimated the necessity of ophthalmologic screening for children with diabetes mellitus. PMID- 19292093 TI - [Hysteroscopy in the diagnosis and treatment of tubal infertility]. AB - The aim of this study is to present the authors' experience in the diagnosis and treatment of tubal proximal disease by means of hysteroscopy. The study includes 96 patients with tubal infertility to which hysteroscopy was performed in 1998 2003 in the 2nd Clinic of Obstetrics and Gynecology, Iasi. The average age was 32.5 +/- 1.2 years. The indication was proximal tubal obstruction diagnosed by hysterosalpingography or sono-hysterosalpingography. The hysteroscopy revealed lesions affecting fertility that were treated in the same operative sequence. Proximal tubal obstruction was present in 50 cases (52%): unilateral--41 cases, bilateral--9 cases. Tubal permeabilization was performed by tubal catheterization or by removing polyps, adhesion and was verified by chromo-tubation at the end of hysteroscopy and by sono-hysterosalpingography, 2 months after the intervention. When proximal tubal obstruction is suspected, hysteroscopy offers a solution by confirming the diagnosis and by efficiently treating the lesions, leading to an increased average of pregnancies. PMID- 19292094 TI - The prenatal detection of Jeune syndrome (asphyxiating thoracic dystrophy). Case report. AB - The authors present the case of a foetus with Jeune syndrome (asphyxiating thoracic dystrophy) in a woman with a previous deceased child with the same disease, and also with a normal sibling. The diagnosis was mentioned at 26 week of pregnancy, based on ultrasonographic findings: short proximal bones (under 3 percentiles), and a diminished thoracic circumference, (although greater than 10 percentiles for the gestational age). There was an interdisciplinary agreement for the therapeutic termination of the pregnancy, and the post-expulsion assessment confirmed the diagnosis. This case demonstrates a higher incidence of Jeune dystrophy than the one expected for an autosomal recessive disease, with 2 out of 3 children affected, instead of 25%. It also shows that the earliest change is the one regarding the shortened long bones, often difficult to notice before 20 weeks, fact which favors a detailed genetic sonogram done after this limit. PMID- 19292095 TI - [Severe peripheral ischemia of rare origin]. AB - The authors describe a rare case of acute peripheral ischemia in a young patient, following a lower limb orthopaedic surgery. The clinical presentation and diagnosis tests suggested a Horton arteritis with peripheral symptoms, although the temporal artery biopsy was negative. Since the etiology was unclear, the authors considered the influence of local trauma (initial and surgical), of Ergot derivatives and antibiotic treatment, but the clinical outcome after corticotherapy strongly indicated a temporal arteritis. PMID- 19292096 TI - [Parathyroid carcinoma in a patient on hemodialysis for renal failure]. AB - Parathyroid carcinoma is a rare endocrine neoplasm with difficult histological diagnostic and unpredictable evolution. More unusual are the cases appearing in the course of end-stage renal disease on maintenance dialysis (22 observations in medical literature). A 46-year-old man suffering of chronic glomerulonephritis and renal failure for 13 years, having been on hemodialysis for three years, complains of asthenia, adynamia, muscle weakness, progressive osteoarticular pains, itching. The preoperative iPTH was 71/ng/ml, seric Ca2+/1.23 mmol/l, seric Ca/2.6/mmoli/l, seric P/2.02 mmol/l. Clinical and ultrasonographical examinations revealed a left "thyroid nodule" of 44 x 37 mm but no images of the parathyroid. Upon surgery, three parathyroid glands (two from the right side and the superior left one) were identified and excised after the frozen section. The fourth gland was not found but the resected "nodule" together with the adjacent thyroid lobe proved a parathyroid carcinoma upon paraffin examination. The clinical course after operation was uneventful. The authors underlined the criteria and difficulties in the pathologic diagnosis of parathyroid carcinoma and also the principles of surgical treatment based on their experience of three cases. PMID- 19292097 TI - [Abdominal gunshot wound with multiple visceral lesions]. AB - The authors present the case of a 21-years-old patient who was admitted in the hospital with a severe abdominal gunshot wound and hemorrhagic shock. Because of the gravity of the external bleeding and the severity of his general status he was admitted directly to the surgery room. The decision of surgical treatment was taken from the first look, so all the diagnostic maneuvers were taken in the surgery room at the same time with the preparations for general anesthesia and surgery, except for the clinical exam and preservation of blood and urine. The duration of the surgical operation was relatively short, almost 2 hours, compared with the visceral lesions identified and treated. The recovery was amazingly rapid, with no significant complications, the patient being discharged from the hospital after 17 days. This case has some particularities which can be discussed: immediate surgery is the only way to give a chance of survival in these kinds of situations; never loose time with other investigations; single gunshot wound of the abdomen determined 4 visceral lesions, every one of them being very severe; the patient had a very short period of recovery. PMID- 19292098 TI - [Hansson Twin Hook--a new implant technique in trochanteric fractures]. AB - Hansson Hook is an implant developed in Scandinavian countries for femoral neck fractures. Due to the good results given by this technique the implant was developed in order to use it in trochanteric fractures. The hook was doubled and the implant adapted to a plate with barrel, similar with those used in compression screw technique. The new implant is named Omega 2 Hansson Twin Hook and is produced by Stryker Osteonics. In the authors' department five patients with trochanteric fractures were treated using Hansson Twin Hook. According to Evans classification, three fractures were stable and two unstable. The surgical technique is somewhat different than a common compression screw implantation: the incision is smaller, the plate is the first to be implanted, the expanding of the hooks requires special instrumentation. Patients were allowed to walk with full weight the day after the surgery. The postoperative results were very good, both immediately and three months after operation. In conclusion, the authors believe that Hansson Twin Hook is a useful implant in trochanteric fractures and offers a good rotational stability. PMID- 19292099 TI - Biosynthesis and physio-pharmacological actions of angiotensin peptides: 1. Synthetic enzymes. AB - The present work introduces a brief review of the actual knowledge concerning the enzymes involved in the biosynthesis of the active angiotensins, followed by a presentation of their main physio-pharmacological actions. The enzymatic pathways that generate active ang. II (1-8) are complemented with data concerning its transformation into angiotensin III (2-8), ang. IV (3-8), ang. V (1-5) and ang. 1 7. Besides the classic renin of renal origin, the tissue isorenins, represented by tonin and cathepsins D and G, inactive angiotensin-I-forming are also reviewed. Furthermore, chymase and the new angiotensin-converting enzyme 2 (ACE2), which generates angiotensin 1-7, having opposite properties from the mother-substance (Ang. II) are discussed at length. The presentation of properties of angiotensin-generating enzymes is followed by the presentation of the action of angiotensinases (aminopetidases, carboxypeptidase and endopeptidases), which are involved both in the generation of biologically active angiotensin peptides and in their inactivation. PMID- 19292100 TI - [Mutagenic aspects of the lipoprotein lipase gene]. AB - Lipoprotein lipase (LPL) is an enzyme involved in the metabolism of triglyceride rich lipoproteins (chylomicrons and VLDL), participating also in the remodeling of HDL particles. It is encoded by a gene located on chromosome 8 (8p22), containing 10 exons. Abnormalities in this gene lead to the development of LPL enzyme deficiency. About 100 mutations have been described in the LPL gene, the most frequent being Asp9Asn, Gly188Glu and Asn291Ser. Mutations in the homozygous form are associated with type I hyperlipoproteinemia (familial chylomicronemia). Mutations in the heterozygous state have a significant incidence in population (3 7%) and lead to a decrease in the LPL activity with up to 50%, which causes the modification of the plasma lipid profile, meaning an increase in triglycerides and a decrease in HDL cholesterol. Both modifications represent cardiovascular risk factors, so that the carriers of LPL gene mutations have an increased predisposition to develop coronary heart disease. Moreover, the association of heterozygous mutations in the LPL gene in individuals with genetic-type hyperlipoproteinemias may aggravate the perturbations of the lipid profile and, consequently, they can increase the cardiovascular risk in these patients. The accumulated data may be considered an argument for the importance of investigating LPL gene mutations in the population, in order to detect precociously the individuals with an increased atherogenic predisposition and to decide on the appropriate therapy. PMID- 19292101 TI - An image pre-processing automatic system for bone scan metastasis evaluation. AB - Scintigraphic images provide morphologic and functional information referring to pointed human body tissues and organs. Depending on the ability and experience of the nuclear medicine physician and the degree of scintigram complexity, the diagnosis process becomes very difficult sometimes. Image processing might decrease subjectivity and help physicians in decision making in complex cases as bone metastasis diagnosis based on scintigraphic explorations. The aim of this paper is to describe the compulsory steps of a pre-processing method in order to build a database for an automatic final appreciation of pathologic bone scan areas as a percentage of the total bone scintigraphic surface. This may include the scintigraphic result in some metastasis probability category with more accuracy than a simple, subjective appreciation of the scintigram, especially in doubtful cases. This paper points to the steps of the processing method of the database used in the rule-based nuclear medicine aide-decision expert system (NMADES). The objective evaluation of the pathological sites requires image preprocessing operations in a number of steps: histogram transforms, correlated superposition of direct and reversed incidences to reinforce the uptake sites, smoothing by pseudo-cepstrum methods, symmetry axes extraction by robust linear regression and symmetric areas search with fuzzy methods. Some for and against's are underlined in the last section, devoted to conclusions and future work. PMID- 19292102 TI - Interactions between angiotensin I and acetylcholine on rat left main bronchial rings. AB - Angiotensin (Ang) II is known to amplified bronchoconstriction induced by acetylcholine (ACh). On the other hand all the components of renin angiotensin system were located on lungs. Contractile effects of Ang I (the precursor of Ang II) and interactions between Ang I and ACh on rat bronchial rings were characterize using angiotensin II type 1 (AT1) receptor antagonist (losartan), angiotensin converting enzyme (ACE) inhibitors (captopril and teprotide) and chymase inhibitor (chymostatin). We found that Ang I has contractile effects and amplified ACh-induced contractions. Blocking of AT1 receptors with 10 mM losartan significantly reduced 10 mM Ang I contractile effects (12.79 +/- 9.59% from 167.62 +/- 8.92%; p<0.05). Pre-treatment with 1 mM teprotide reduced 10 mM Ang I induced contractions (35.68 +/- 7.83%; p>0.05). Captopril and teprotide only reduced Ang I actions. This suggested that both types of Ang I effects were mediated by AT1 receptors, but possibly conversion of Ang I into Ang II were not significantly dependent by ACE or chymase. PMID- 19292103 TI - [The influence of the dihydroxamic compounds on the rat lung]. AB - The role of the dihydroxamic compounds in the human and animal organism are little known. This is the reason they the experiment studies the effect of these compounds on the rat lung. 3 lots of Wistar adult male rats were used. The first lot was the witness lot and did not receive any substance. The second lot received Dihydroxamic adipoil acid, 0.5 mg/kg weight/ day, intraperitoneal administration in a unique daily dose, for 3 weeks. The third lot received Dihydroxamic terephtaloyl acid, 0.5 mg/kg weight/day, intraperitoneal administration in a unique daily dose. After 14 and 21 days the animals of each lot were euthanised by sectioning carotid arteries. The authors collected the lungs, prepared the histological sections, stained in HE and Szekely. The witness lot presented normal rat lung structure. After 2 and 3 weeks of treatment, the second lot presented distended blood vessels. After 2 and 3 weeks of treatment, the third lot presented very severe alterations of the lung tissue at all levels. PMID- 19292104 TI - [Nutrition and periodontal disease]. AB - It is difficult ro assess the role of nutrition in the etiology and progression of periodontal disease because many other factors besides the local effect of plaque affect periodontal tissue metabolism. It is clear that nutrition can affect host response to bacterial plaque and it is also apparent that there may be a need for the intake of greater amounts of certain nutrients (such as ascorbic acid, iron etc.). Inadequate nutrient intake or deficiency is significant because of the number of interactions that occur during the assimilation of foods and the effects of stress and medication. Periodontal health cannot be achieved unless nutrient deficiency is corrected along with the other phases of treatment. PMID- 19292105 TI - Periosteoplasty--a new technique for recession coverage. AB - A new technique of periodontal flap surgery was performed on 5 patients with severe periodontal recessions of the upper or lower anterior teeth. The procedure begins with a thorough root and soft tissue scaling. For the surgical approach the periosteum was incised and mobilized at the apical part of the mucoperiosteum flap; it will further be used to cover the defect. The mucoperiosteum flap is then readapted and sutured. Prior to surgery the following clinical parameters were registered: sulcus bleeding index, periodontal probing depths, attachment loss and the height of the attached gingiva. Registrations of the same clinical features were made 3, and 12 months after surgery. Each clinical parameter registered an improvement following surgery. No sulcus bleeding was observed at any time during the postoperative follow-up. The evaluation after 12 months showed an average of 5.5 mm gingival reattachment and an average of 0.3 mm probing depth. The mean height of the attached gingiva was 0 mm before surgery, 2.3 mm at the 3 and 6 month evaluations and 2.2 mm after 12 months. The periosteum eversion technique is suitable for the treatment of gingival recessions, resulting in good gingival functions and the obvious improvement in appearance. PMID- 19292106 TI - [Root caries prevalence in Romania. Study in Oral Health Center, Iasi]. AB - Root caries are a frequent pathology with an evolution compromising the vitality and even the persistence of the tooth on the arch. The WHO study offers information about the prevalence of those lesions and the treatment needs, orienting on the prophylactic measures and therapy. The study evaluated 10,415 patients, revealing some deficiencies in the interfering of the evolution of those lesions. The most affected teeth were the inferior canines and incisors and the superior third molars. The study recommends a focused prophylaxis concerning also the elderly people. PMID- 19292107 TI - [Evaluation of caries activity using the micro-crystallization saliva index (IMK)]. AB - The dental decay activity of a patient can be evaluated with the help of certain indirect signs that reveal the dental decay risk elements. The quantity, buffer capacity, inorganic components of the saliva are important in the pathogenesis of dental decay. The micro-crystallization saliva test (IMK) may represent an important criterion on whose basis one obtains an evaluation of the re mineralization capacity of saliva. The purpose of this study consisted in the calculation of the IMK index on a lot of patients and its correlation to the type of dental decay activity. The study was conducted on a lot of 30 patients with ages between 18 and 45, nursed and examined every 6 months during 18 months. The attention was focused on the following parameters: the index of micro crystallization (IMK) and the dental decay activity, calculated as rate of appearance of new dental lesions within 18 months. As a result of investigations, the authors observed the following correlations to dental decay activity: the patients with IMK = 1/0.6 did not have any new dental lesion, in the patients with IMK = 0.6/0.4 new dental lesions appeared within 12-18 months and in the patients with IMK = 0.4/0 new dental lesions appeared in less than 6 months after examination. The IMK index--indicator of the re-mineralization capacity of saliva, may be evaluated using a relatively simple methodology, accessible and at a moderate price. Research pointed out that this index may be an useful instrument when evaluating the dental decay susceptibility of patients, contributing to the identification of the dental decay risk and consecutively to the choice of individualized prophylactic and therapeutic procedures. PMID- 19292108 TI - [The distribution of anaerobic morpho-types in sub gingival plaque associated with periodontal disease]. AB - The authors' goal has been to establish the distribution of anaerobic morpho types in sub gingival plaque collected from patients with periodontal disease. The study included 26 patients. Samples of sub gingival plaque were transported in thioglycolate broth. The authors did Gram stains and culture on three types of media. The plates were incubated at 37 degrees C in anaerobic atmosphere for 5-7 days. From colonies with growth of at least +2 Gram stains and tolerance to oxygen were studied. The authors isolated 156 strains (an average of 6 strains per patient). 134 strains were anaerobic bacteria. Anaerobic morpho-types were distributed as follows: 70.01%--gram negative bacilli, 11.19%--gram positive cocci, 10.44%--gram negative cocci, 8.20% gram positive bacilli. Gram negative anaerobic bacilli were present in all samples and in 80% cases they were associated with other bacteria. PMID- 19292109 TI - [Research concerning the characterization of Allium cepa based ointments]. AB - Three topical formulae with Allium cepa L. extract 30% (w/w) were studied for the evaluation of their release profiles of flavonoids and for their potency on experimental wounds on mice. The rates of release were performed under occluded conditions using cellulose acetate membrane. Data obtained from the in vitro release show that the gel formula F1 is the most efficient in the membrane diffusion process. The rate of wound healing was assessed by the contracting ability and the period of epithelization. The most important effect in the contracting ability was noticed in the case of F2 formula. PMID- 19292110 TI - [The extraction and transport of folic acid through liquid membranes]. AB - The study on the extraction and transport of folic acid (vitamin B9) with Amberlite LA-2 underlined the significant influences of the pH-gradient on the aqueous phases, carrier concentration and mixing intensity, in correlation with the extraction mechanism. Thus, for reaching the maximum efficiency of folic acid separation, the pH-value of the initial aqueous phase of 5, the pH-value of the final aqueous solution greater than 11 and the Amberlite LA-2 concentration in solvent layer over 80 g/l are required. PMID- 19292111 TI - [Synthesis and physico-chemical characterisation of some new derivatives of rutoside and clofibric acid]. AB - Fibrates are drugs with efficacy in reducing blood cholesterol levels and especially, triglyceride plasma levels. Unfortunately, fibrates have a poor water solubility and showed some adverse reactions at long treatment. The objective of this study was to obtain some new clofibric acid derivatives with rutin; some of these compounds contain a guanidine moiety, known as effective at cardiovascular level. All the compounds are soluble in water. PMID- 19292112 TI - Synthesis and characterization of a hydrogel with therapeutic applications. AB - The study presents the synthesis of an acrylamide polymeric hydrogel in aqueous solution by copolymerization concomitant with a crosslinking process using a bifunctional co-monomer with a redox initiation system--potassium peroxo disulphate/ascorbic acid. The swelling kinetics as well the characterization of structure through optical microscopy was performed. The synthesized polymeric gel shows temperature sensitivity, maintaining low temperatures for a prolonged period of time. Thus, the material is assessed as an inner material in thermo sensitive packs to be applied in pain therapy. PMID- 19292113 TI - Stress and deformation analysis induced by dental implants in mandible. AB - The paper presents the human implanted mandible behavior under the action of muscle forces creating different stress distributions depending on the kind of motion performed. The mandible is treated in accordance to the classical mechanics methods. The model is based on the mandible actual shape, taking into account the correct geometric parameters. By imposing the motion conditions during mastication and by considering the real forces developed by muscles during this motion, the reaction forces acting in the temporal-mandible joint and on each tooth have been computed. The aim of the paper is to establish a connection between the implant number and the jaw mechanical behavior. PMID- 19292114 TI - [Natural zeolites with medical applications--preliminary preparation and characterization]. AB - The aim of this research is to carry out a preliminary study of the chemical behavior of natural zeolites under thermal and chemical activation. Taking into account the biological properties reported for the NZ, the authors study the influence of different activations on the structural behavior of the raw material, aiming at the future preparation of slow release systems. The raw material used in this study was the purified natural zeolites from Barsana, Piatra Verde and Slanic Moldova deposits. The samples were characterized following specific techniques: XRD, SEM, EDAX, IR, pH measurements and determinations of the specific surface. The results indicate that the natural zeolites could used like as carriers in medical applications because they increase their specific surface after activation. Studies of the release of pharmaceuticals from the zeolitic host systems are in progress. PMID- 19292115 TI - Muddy water. PMID- 19292116 TI - A rare case of signet ring cell gastric adenocarcinoma. AB - Although the incidence and mortality rates continue to decline in the United States, gastric cancer remains one of the leading causes of cancer mortality worldwide. Signet Ring Cell gastric cancer is a rare and distinct histological subtype. This is a case report of an individual who presented with nausea, vomiting, abdominal pain and weight loss. The patient was found to have a large, obstructing antral mass that was biopsied by esophagogastroduodenoscopy (EGD) and revealed Signet Ring Cell gastric adenocarcinoma. Preoperative staging with positron emission tomography (PET) and computed tomography (CT) scan displayed multiple pulmonary nodules felt to be consistent with sarcoidosis after transbronchial biopsy results were obtained. Patient was taken to surgery for curative resection and lymph node dissection, but peritoneal metastatic disease was found intra-operatively. Gastric carcinoma frequently requires a multi disciplinary approach to properly diagnose, stage, and treat. PMID- 19292117 TI - MSMA: physicians caring for Mississippi. PMID- 19292118 TI - Congressional tampering with prescription pad requirements: vexation without representation? PMID- 19292119 TI - "Give 'em a bullet to bite on": Civil War medicine and Mississippi's medical heritage. PMID- 19292120 TI - Terminating the patient-physician relationship. Abandonment of the patient: what it means and how to avoid it. PMID- 19292121 TI - The impact of Hurricane Katrina upon older adult nurses: an assessment of quality of life and psychological distress in the aftermath. AB - The primary purpose of the current study was to evaluate the impact of Hurricane Katrina upon older nurses using cross sectional data from 291 respondents. Collected data served as the numerical predicate for the evaluation of quality of life and psychological distress among nurses who were affected by Hurricane Katrina. While the focus for the present study was upon older nurses, cross sectional data was reflected for the plenary sample as well. Predictors of Katrina's impact upon older nurses were identified through multinomial regression analyses and included the physical function subscale (OR=0.954), the fatigue subscale (OR=0.961), the arousal subscale (OR=4.190), average to poor health (OR=2.040), married (OR=2.769) and the MSPSS (OR = 0.780). Significant associations between age and storm impact (F=10.707, n=.001), depression (F=15.782, n< .001), social support (F=5.869, n=.016), health status (F=29.004, n<.001), anxiety (F=5.583, n=.019) and posttraumatic distress disorder (F .032, fn= .46) remained after adjustment for other risk factors. These associations, as reflected in their respective mean scores, indicated that older nurses experienced greater storm impact (2.880 vs. 2.511), depressive symptoms (11.250 vs. 9.080), anxiety (77.800 vs. 75.430), posttraumatic distress (72.830 vs. 70.860) and lower health status (68.891 vs. 73.569). Accordingly, a more robust public policy paradigm for addressing the growing labor shortages in the medical community is needed. Heightened Congressional interest and increased resourcing is required in order to affect necessary programmatic, educational and institutional remediation. Furthermore, given the increasing role of older nurses in the work place, extensive studies are needed to evaluate their status and independent risk factors for sustaining quality of life and psychological well being among these contributors of health care. PMID- 19292122 TI - More than meets the eyes. PMID- 19292124 TI - Wanna hear something really scary? (Some thoughts on children's health). PMID- 19292125 TI - No turning back. PMID- 19292126 TI - A time to heal. PMID- 19292127 TI - Hospital payment monitoring program concludes. PMID- 19292128 TI - UMC neurosurgery patient regains function despite virtual decapitation. PMID- 19292129 TI - Images in Mississippi medicine. Quarantine against hydrophobia. PMID- 19292130 TI - Stuff that wears you out. PMID- 19292131 TI - Indications for pediatric cochlear implantation at the University of Mississippi Medical Center. AB - OBJECTIVE: To classify and describe causes of hearing loss in children of Mississippi. SETTING: Tertiary, university practice. DESIGN: Consecutive series of children undergoing cochlear implantation at the University of Mississippi Medical Center. MATERIALS AND METHODS: A prospectively maintained database has been kept since the inception of the cochlear implant program at UMMC. Records were queried as to the diagnosis of hearing loss, perioperative complications, and postoperative acceptance of the device by the patient. RESULTS: Fifty-nine children underwent cochlear implantation from April 2003 to March 2007. Twenty two children had no risk factors for hearing loss and were presumed to have genetic hearing loss. Seven had hearing loss related to complications of prematurity. Four had abnormal genes for Connexin 26, and four had congenital cytomegalovirus infections. Three had unclassified autosomal dominant hearing loss. CONCLUSIONS: Since nearly half of the patients in this series had no known risk factors for hearing loss, our findings support the need for universal newborn hearing screening. Further advancements in genomic research will facilitate diagnosis of genetic hearing loss. PMID- 19292132 TI - Strange bedfellows. PMID- 19292133 TI - Statewide coordination of Mississippi Health Awareness Day. PMID- 19292134 TI - There are giants in the land. PMID- 19292135 TI - Mississippi priorities. PMID- 19292136 TI - Eggs, beaches and shotguns. PMID- 19292137 TI - Carisoprodol abuse in Mississippi. PMID- 19292138 TI - A case of severe hemorrhagic diarrhea. AB - A 50-year-old white female presented to the emergency department with diarrhea, abdominal pain, and hematochezia. The illness started four days prior to presentation and escalated to 10-15 bowel movements daily. She was tachycardic and hypotensive upon presentation with a diffusely tender abdomen. Her anemia and persistent hematochezia prompted endoscopy by a gastroenterologist. Flexible sigmoidoscopy was performed to the splenic flexure and revealed evidence of severe active colitis. The mucosa was friable and severely ulcerated. Numerous biopsies were obtained that revealed acute colitis with fibrinopurulent exudates. A stool culture showed Escherichia coli O157:H7, which was confirmed by the Mississippi State Board of Health. Infection with E. coli O157:H7 affects nearly 1.1 per 100,000 Americans annually. It accounts for about 3% of all bacterial and protozoal causes of foodborne illnesses and possesses a mortality rate of about 1 2%. PMID- 19292139 TI - Special six-county coast project implemented after Hurricane Katrina. PMID- 19292140 TI - My Christmas wish list. PMID- 19292141 TI - Paget's sarcoma of the radius. PMID- 19292142 TI - Fibular autograft and silicone implant arthroplasty following resection of giant cell tumor of the metacarpal: a case report with 8 years follow-up. PMID- 19292143 TI - Synchronous multicentric osteoid osteoma with associated fibrous cortical defect. PMID- 19292144 TI - Isolated lesser tuberosity fracture of the humerus. PMID- 19292145 TI - Medial elbow dislocation associated with a fracture of the lateral humeral condyle in a child. PMID- 19292146 TI - Association of Streptococcus bovis haematolgenous infection in total knee replacement with large intestine carcinoma. PMID- 19292147 TI - Tibia vara secondary to focal fibro-cartilaginous dysplasia of proximal tibia. PMID- 19292148 TI - Patellar clunk syndrome after TKA with cruciate retaining design: a report of two cases. PMID- 19292149 TI - Permanent loss of cervical spinal cord function after posterolateral fusion for lumbar spinal pyogenic spondylitis. PMID- 19292150 TI - Complete quadriceps tendon rupture with concomitant tears of the anterior cruciate ligament and lateral meniscus. PMID- 19292151 TI - Rupture of anterior cruciate ligament associated with fractures of ipsilateral femoral shaft and neck. PMID- 19292152 TI - Peroneal nerve palsy secondary to ganglion cyst of the proximal tibiofibular joint in a pediatric patient. PMID- 19292153 TI - Primary intermuscular hydatid cyst of the thigh. PMID- 19292154 TI - Osteochondroma development subsequent to traumatic hip dislocation. PMID- 19292155 TI - Septic arthritis of the hip in relation with femoral neck fracture: a report of three cases. PMID- 19292156 TI - Dissociation of modular total hip arthroplasty at different levels due to subsidence of cementless stems. A report of three cases. PMID- 19292157 TI - The presence of bilateral hip dysplasia and genu valgum in Fraser syndrome. PMID- 19292158 TI - Use of a stacked Galveston functional brace for treatment of multiple adjacent metacarpal fractures. PMID- 19292159 TI - Marginal excision of squamous cell carcinomas of the hand. AB - Squamous cell carcinoma is the most common malignant tumor of the hand. Most hand surgery texts recommend wide excision with 1- to 2-cm margins. During a 10-year period, 32 patients with 35 lesions were treated with marginal surgical excision for squamous cell carcinoma of the hand. Greater potential morbidity and functional loss associated with wide excision was avoided in these patients. No recurrences were found at a mean follow-up of 31 months. One case of possible metastasis was identified. Three (8%) tumors required further treatment due to positive surgical margins. Marginal excision of squamous cell carcinoma of the hand is associated with a low rate of recurrence and metastasis. In addition, the potential for improved cosmesis and diminished morbidity when compared to older more radical surgical methods supports marginal excision for squamous cell carcinoma of the hand as a viable treatment technique. PMID- 19292160 TI - Management of nonunion associated with broken intramedullary nail of the femur. AB - This study analyzed 14 patients with 17 broken intramedullary nails for the treatment of femoral fractures. Average distance from the fracture site was 7.9 cm in cases in which nail breakage occurred at the junction between the cylindrical and cloverleaf portions and 6.4 cm in cases in which nail breakage occurred at the screw hole. Eleven patients with 14 broken nails were treated with exchange nailing and bone grafting, 2 were treated with plating and bone grafting, and 1 was treated with extracorporeal shock wave therapy. Nonunion or delayed union is the main cause of nail breakage. Exchange nailing with bone grafting is a safe and effective method of treatment for a broken intramedullary nail with nonunion. PMID- 19292161 TI - Screw osteolysis in the cementless anatomic modular knee arthroplasty. AB - A cohort of patients underwent prospective follow-up to delineate the natural history of screw-associated osteolysis in cementless Anatomic Modular Knee arthroplasty. In 1993, fluoroscopically guided radiographs were obtained in 230 patients (280 arthroplasties) to identify occult osteolysis. This cohort was reassessed periodically to identify new or progressive screw-associated osteolysis. At early follow-up, 94 knees (34%) had osteolysis. Osteolysis progressed in all groups, and osteolysis developed in 60 additional knees. At intermediate follow-up, 55% of knees showed radiographic evidence of osteolysis. Osteolysis progressed to a higher grade in a significant percentage of patients, and 23% of arthroplasties were revised. Regular periodic radiographic evaluation is recommended for early recognition of osteolysis. PMID- 19292162 TI - Trend of the carrying angle during flexion-extension of the elbow joint: a pilot study. AB - This study evaluated the variability of the carrying angle during flexion extension of the elbow and defined a description of the movement applicable in sport and rehabilitation.The arm and forearm of 7 healthy volunteers were marked and the coordinates were acquired during motion using 6 infrared cameras. Five repeated measures were performed for each volunteer to verify the reliability.Our results demonstrated that this movement can be easily modeled as a linear variation of the carrying angle in function of the flexion angle.The reliability between repeated measures was high and adopting a linear fit the accuracy was >94%. PMID- 19292163 TI - Proximal row carpectomy with capitate resection and capsular interposition for treatment of scapholunate advanced collapse. AB - Proximal row carpectomy with capitate head resection and dorsal capsular interposition was performed in 8 patients with stage II and III scapholunate advanced collapse (SLAC). Mean patient age was 58.5 years (range, 50-79 years). One year postoperatively, range of motion and grip strength were not significantly changed from preoperative values. Mean postoperative Disabilities of the Arm, Shoulder, and Hand questionnaire score was 27. Pain improved from 8.8 preoperatively to 4.3 postoperatively on a scale of 0-10. Pain levels were highly inversely correlated with final range of motion (r=-0.94, P=.0004). Proximal row carpectomy with capitate leveling and capsular interposition for stage II and III SLAC wrist provides reasonable pain relief in 75% of patients. Patients should be counseled that 25% of cases may demonstrate progression of radiocapitate degeneration, necessitating further intervention. PMID- 19292164 TI - Arthroscopically assisted mini-deltopectoral rotator cuff repair. AB - This study examined the technique and outcome of arthroscopically assisted mini deltopectoral rotator cuff repair. The mini-deltopectoral approach enables the repair of large or massive rotator cuff tears while preventing injury to the deltoid muscle and axillary nerve often caused by splitting and excessive traction during conventional mini-open repair. This technique also is useful in the repair of subscapularis tears and thus could extend the indications for mini open repair. PMID- 19292165 TI - Osteoarticular autograft and allograft transplantation of the knee: 3 year follow up. AB - Between 1998 and 2001, a total of 48 patients underwent autologous or allograft osteoarticular transplantation with a minimum follow-up of 24 months. Nineteen patients underwent concomitant procedures. Average patient age was 46 years. Graft area averaged 4.8 cm2 (range, 0.2-21.6 cm(2)). Grafts implanted included 24 autografts, 12 refrigerated allografts, and 12 frozen allografts. Average follow up was 37.1 months (range, 24-63 months). Postoperative Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, Knee Society Score (KSS), and pain scores improved significantly. Age <35 years and male gender was associated with a positive outcome. There was no significant difference in improvement between autograft and allograft groups. Nine patients, considered failures, underwent total knee arthroplasty and data analysis. No significant relationship was noted for any variable. Eighty-one percent of grafts are still functioning >3 years after transplantation. PMID- 19292166 TI - Clinical application of rapid prototype model in pediatric proximal femoral corrective osteotomy. AB - Proximal femoral deformities can occur in children as sequelae of several disorders such as hip joint infection or avascular necrosis of the femoral head. Early surgical correction of deformities promises a good result; however, limited exposure of the surgical field often creates difficulties for surgeons. Application of a rapid prototyping model for proximal femoral corrective osteotomy in children serves not only as a guide for surgical planning but also as a communication tool between physicians and patients. PMID- 19292167 TI - A retrospective analysis of treatment and outcomes in patients with late onset tibia vara. AB - This retrospective analysis included 36 patients who presented with late-onset tibia vara during a 15-year period. Mean patient age at presentation was 12.8 years. Seventy-eight percent of patients were male, 78% were black, and 72% presented with unilateral deformity. Mean initial varus was 21 degrees with 5.8 degrees at the femur and 12.3 degrees at the tibia. Surgical correction was performed in 31 patients; 5 patients refused surgery. Patients who had osteotomies had a higher rate of complications than those treated with stapling. Epiphyseal stapling has unpredictable results in patients with late-onset tibia vara and is only a temporizing procedure in approximately half of cases. None of the treatments resulted in optimal knee function or alignment, and the long-term prognosis for late-onset tibia vara is guarded at best. PMID- 19292168 TI - Evaluation of a tibial torsiometer applicable to clinical practice. AB - This study evaluated a new tibial torsiometer that is universally applicable to all limb sizes and requires no special training, jigs, or radiographic equipment. To compare the reliability of measurements obtained with the torsiometer, 160 tibias were evaluated with both the torsiometer and computed tomography. Results for both methods were identical in 51 tibias, within 1 degrees in 66 tibias, and within 2 degrees in 43 tibias. The difference between the two methods was not significantly different. No significant difference was found among testers. This device should prove to be useful for the quantitation of tibial torsion. PMID- 19292169 TI - Functional outcome in adult patients following Bernese periacetabular osteotomy. AB - This study evaluated functional outcome following Bernese periacetabular osteotomy. In 24 patients with mean follow-up of 3.5 years, mean dysfunction score was 15.23 on the Short Musculoskeletal Function Assessment. Eighteen patients (75%) scored = or <20, indicating a high functional level. Irrespective of preoperative Shenton line continuity, most patients demonstrated a high functional level. However, a trend toward a poorer outcome was observed in patients with preoperative noncongruent joints and Tonnis osteoarthritis grade 3. These results suggest patients with less than optimal presentation may still benefit from this surgery, delaying or eliminating the need for total hip arthroplasty. PMID- 19292170 TI - Changes of the posterior meniscal horn height during loading: an in vivo magnetic resonance imaging study. AB - This in vivo magnetic resonance imaging study evaluated the deformation of the meniscal posterior horn during loading. Images of the medial and lateral meniscus were obtained in 20 normal knees in 0 degrees and 30 degrees of flexion, and the height of the posterior horn of both menisci was measured. Mean height of the posterior horn of the medial meniscus under no load and load equal to 50% and 100% body weight was 6.8, 6.3, and 6 mm, respectively (P<.05). Mean height of the posterior horn of the lateral meniscus under no load and load equal to 50% and 100% body weight was 7.6, 7.2, and 6.9 mm, respectively (P<.05). Knee position (0 degree versus 30 degrees of flexion) at a constant load resulted in no significant change in height. These results show height of the posterior horn of the medial and lateral meniscus decrease with increasing loading and angle of knee flexion. PMID- 19292171 TI - Extended femoral osteotomy and proximally coated prosthesis for hip revision. AB - Extended trochanteric osteotomy facilitates femoral reconstruction in revision total hip arthroplasty. This study examined the outcome of extended trochanteric osteotomy combined with a proximally porous-coated calcar replacement prosthesis during revision total hip arthroplasty. Forty-five cases were reviewed with an osteotomy union rate of 98%. No femoral component was loose radiographically, and none were revised for loosening. These findings indicate excellent short-term clinical and radiographic results can be achieved with an extended trochanteric osteotomy and proximally porous-coated calcar replacement prosthesis. PMID- 19292183 TI - Synovial lipomatosis of the knee in an adolescent girl. AB - Synovial lipomatosis is an uncommon intraarticular lesion rarely affecting the pediatric age group. Also known as Lipoma Arborescens (LA), it is characterized by extensive villous proliferation of the synovial membrane and hyperplasia of subsynovial fat. The etiology of this condition remains unknown. It should be considered in the differential diagnosis of refractory chronic joint effusion and synovial swelling around the knee joint. MRI is helpful in the diagnosis of this pathology. Treatment is by arthroscopic excision. We report the findings in a 15 year-old girl who presented with a painful, swollen knee of 4 years duration. She was found to have Synovial Lipomatosis of the knee joint, which was excised arthroscopically. Synovial lipomatosis is a rare diagnosis in childhood. It should be considered in the differential diagnosis of refractory chronic joint effusion and synovial swelling around the knee joint. Magnetic resonance imaging may be helpful in the diagnosis of these cases, but confirmation can only be obtained through histology. The recommended treatment is arthroscopic excision of the lesion. In recurrent cases, open synovectomy may be required. PMID- 19292184 TI - Acute compartment syndrome of the calf presenting after prolonged decubitus position. AB - Acute compartment syndrome subsequent to high velocity trauma and motor vehicle accidents causing fractures and significant soft trauma in the extremities has been reported in the literature. Compartment syndrome also has been reported to occur after prolonged decubitus position while in the lithotomy position in gynecological patients as well in arthroscopic patients. This case report presents a new and different presentation of acute compartment syndrome. A 39 year-old woman presented with severe pain in her right calf after falling off of a sofa. The patient most likely caused extensive compression of the leg compartments by her body weight and position. This case report is a good example of the high level of suspicion that an orthopedists needs to have when a patient presents to the emergency room with severe pain of an extremity after what may initially be considered to be insignificant trauma. PMID- 19292185 TI - Bilateral osteonecrosis of the femoral head associated with pregnancy: four new cases and a review of the literature. AB - Osteonecrosis is thought to result from the temporary or permanent loss of blood supply to the bones. Without proper nutrition, the bone tissue dies, is resorbed, and collapses. If the process involves the bones in a joint, it often leads to collapse of the joint surface. Four new cases of bilateral osteonecrosis of the femoral head occurring during pregnancy are described, with pathologic examination of core tissue biopsy substantiating the diagnoses. Operative and nonoperative management included anti-inflammatory medication, pulsating electromagnetic fields, core decompression, and total hip replacement. A review of the literature is undertaken to document the reported cases of osteonecrosis of the femoral head associated with pregnancy. Idiopathic osteoporosis of the femoral head is a condition that mimics osteonecrosis of the femoral head and can be a challenge in diagnosis during the latter stages of pregnancy. Bilateral osteonecrosis of the femoral head during pregnancy is rare. Patients may have other predisposing factors leading to femoral head necrosis. Transient osteoporosis of the femoral head during pregnancy can present with a clinical picture similar to osteonecrosis. Avascular necrosis of the femoral head should be considered in the differential diagnosis of pain about the hip developing during pregnancy or in the immediate postpartum period. PMID- 19292186 TI - Giant anterior sacral meningocele presenting as bacterial meningitis in a previously healthy adult. AB - Meningocele may be asymptomatic and incidentally discovered. Presenting as a retrorectal mass, sacral meningocele may produce urinary, rectal, and menstrual pain. Anterior sacral meningocele may be the cause of tethered cord syndrome. This article presents a case of a previously healthy 39-year-old man with large meningeal herniation that occupied the entire pelvis who developed symptoms of bacterial meningitis. A 39-year-old man was admitted with fever, chills, headache and photophobia. Escherichia coli was isolated from cerebrospinal fluid culture. Moderate improvement regarding meningeal symptoms was noted due to intravenous antibiotic therapy, but intense pain in the lower back associated with constipation, fecal and urinary incontinence, and saddle anesthesia developed. Abdominal ultrasound was negative. Plain radiographs and computed tomography demonstrated sacral bone defect and retrorectal expansive mass. MRI confirmed anterior sacral meningocele with cord tethering. After posterior laminectomy and dural opening, communication between meningocele and intrathecal compartment was obliterated. Computed tomography-guided percutaneous drainage through the ischiorectal fossa was performed to treat residual presacral cyst. Delayed diagnosis in our patient was related to misleading signs of bacterial meningitis without symptoms of intrapelvic expansion until the second week of illness. In our patient, surgical treatment was unavoidable due to resistive meningitis, acute back pain, and symptoms of space-occupying pelvic lesion. Neurosurgical approach was successful in treatment of meningitis and neurological disorders. Computed tomography-guided evacuation of the residual retrorectal cyst was less invasive than laparotomy, resulting in normalization of defecation and miction despite incomplete evacuation. Further follow-up studies may provide insight into the most effective treatment of such conditions. PMID- 19292187 TI - Supracondylar correction osteotomy to prevent repetitive posterior dislocation of a hip prosthesis. AB - Recurrent dislocation of a total hip prosthesis can be a challenging and often disappointing problem. This case report describes a 78-year-old woman who had recurrent posterior dislocations of a revision total hip replacement (THR) that occurred on flexion and internal rotation of the hip. The initial THR for posttraumatic osteonecrosis and extensive revision THA were performed through an anterolateral approach. Conservative treatment was not successful in preventing posterior re-dislocation, neither was open surgery in which offset was improved. Investigation under anesthesia by means of an image-intensifier showed that the prosthesis was easily dislocated posteriorly on flexion and internal rotation. On external rotation the hip was completely stable. Flexion-internal rotation is a rare motion which, if the patient is clearly instructed should be avoidable. The mechanism of an anterior dislocation caused by external rotation, extension and adduction is more common, especially since an anterolateral approach was used in all procedures. We describe the surgical technique of performing an supracondylar external rotating osteotomy of the femur to avoid further dislocation. Fixation was performed with a condylar plate fixation of which the proximal part is fixated above the distal point of the femur component to avoid a stress riser. Since the foot and knee of the patient are now internally rotated, the patient is forced to produce an external rotation in the hip to have a normal leg alignment, therefore preventing the internal rotation on which the dislocation occurred. In the prevention of a re-dislocation, solutions other than performing a revision of the THA can successfully solve the problem and should therefore be considered. PMID- 19292188 TI - Bilateral femoral neck fractures in long-term steroid use for systemic lupus erythematosus and chronic lung disease. AB - Bilateral femoral neck fractures present a rare injury. Only one report to our knowledge was not related to an acute severe traumatic event, and developed years after pelvic irradiation. Chronic steroid use may severely decrease bone strength, thus increasing the risk for such an injury. Patients with chronic lung disease and chronic inflammatory conditions are frequently treated with steroids such as prednisone at doses that may exceed 2.5 mg a day for long durations. Fractures at vulnerable sites such as the femoral neck may then follow without any severe trauma. Awareness of the detrimental effect of chronic steroid consumption on bone morphology, and familiarity with treatment alternatives to improve bone mass is important to prevent such a severe injury. We describe two cases of bilateral femoral neck fractures in women who were treated for years with orally administered prednisone. The rarity of such an injury of bilateral hip fractures and the fact that neither of the patients sustained major trauma, strongly suggests that both cases were related to impaired bone metabolism due to the effect of prolonged steroid consumption. The biological effects of different roots of steroid administration on bone turnover, as well as several strategies that can be implemented by clinicians to treat and prevent steroid induced osteoporosis and fractures, are further clarified in this article. PMID- 19292189 TI - Total hip arthroplasty in paralytic dislocation from poliomyelitis. AB - This article presents a case of a patient with degenerative hip disease in paralytic dislocation by poliomyelitis. Poliomyelitis is an acute infection disease caused by a group of neurotrophic viruses, which has a special affinity by the anterior horns cells of the spinal cord and for certain motor nuclei of the brain stem. Paralysis is a flaccid type and characteristically paralysis is asymmetrical. It is said that the joints of the affected limb by poliomyelitis are protected from the development of osteoarthritis. Hip dislocation in poliomyelitis is an acquired deformity caused by flaccid paralysis and the resulting muscular imbalance. In young children, when the gluteus maximus and medius muscles are paralyzed and the hip flexors and adductors are of normal strength, eventual luxation of the hip is almost inevitable. Hip osteoarthritis in a limb with poliomyelitis is an unusual entity because these limbs do not support excessive loads. In patients who present with the residual effects of poliomyelitis including degenerative disease and hip dysplastic, surgery is one of the most difficult challenges faced by reconstructive surgeons. In such cases, surgeons should attempt to optimize the component position and choice, surgical approach, and soft tissue tensioning because stability of the prosthesis can be problematic. PMID- 19292190 TI - Femoral nerve palsy following iliacus hematoma. AB - We report a rare case of femoral nerve palsy secondary to hematoma compression of the iliacus. The patient presented with pain 2 days after performing a double kick in karate. The femoral nerve symptoms (numbness and weakness) did not begin until 3 to 4 days after the initial trauma. Magnetic resonance imaging (MRI) confirmed that there was a hematoma compressing the iliacus (and femoral nerve). This hematoma was subperiosteal, within the iliac fossa. This case is rare because the femoral nerve palsy resolved without surgical intervention, the mechanism of injury, the imaging used to diagnose the iliacus hematoma, and the hematoma was subperiosteal and not within the actual iliacus muscle. A review of the literature regarding femoral nerve palsy secondary to traumatic iliacus hematoma is presented. If the diagnosis of femoral nerve palsy secondary to iliacus hematoma is suspected after history and physical examination, MRI is the imaging study of choice. If MRI does not depict a discreet hematoma mass, nonoperative management should be considered. Electromyography has not been shown to offer information that was not provided by history, physical examination, and other imaging. Surgery should be considered if there is progression of the femoral nerve symptoms, or if MRI depicts a discreet collection of blood impinging on the femoral nerve. PMID- 19292191 TI - Autogenous osteochondral transfer in the femoral head after osteonecrosis. AB - Osteonecrosis of the femoral head is a major problem, especially in younger patients. Most of them need a total hip replacement. This article presents a case of a successful autologous osteochondral transfer. PMID- 19292192 TI - Recurrent benign solitary intraosseous schwannoma of the tibia. AB - Schwannomas (neurilemmomas) are benign neoplasms composed of well-differentiated Schwann cells and are usually found in the soft tissues. The occurrence of schwannomas in bone is rare, accounting for less than 0.2% of primary bone tumors. Most cases of osseous schwannoma reported in the world medical literature involve bones of the skull. Only 2 cases have previously been described in the tibia, neither of which has recurred. We report the first case of a recurrent benign solitary intraosseous schwannoma of the tibia, and detail the clinical, radiological and histological findings. PMID- 19292193 TI - Delayed diagnosis of synovial sarcoma of the foot. AB - Soft tissue sarcomas are relatively rare in the population, rarely encountered by the average orthopedic surgeon, and may only be seen once in a lifetime by a general practitioner. Although rare, it is a serious condition that, if diagnosed early, can result in improved prognosis. This article presents a case of a delayed diagnosis of a soft tissue sarcoma and highlights possible pitfalls and causes for such delays in hope of educating and reducing such incidences in the future. PMID- 19292194 TI - Suicide attempt with injection of insecticide in both wrists. AB - Organophosphates that are commonly used in agriculture, houses, gardens, and in veterinary medicine worldwide, may be used for suicidal purposes. But suicide attempt with self-injection of organophosphates is rare. This article presents a case of a suicide attempt of a young man with self-injection of an organophosphate insecticide (dichlorvos) to both his wrists. PMID- 19292195 TI - Brachial plexus neuroma mimicking local recurrence of malignant mesenchymal tumor after forequarter amputation. AB - A stump neuroma is an attempt for the repair of a nerve following amputation. This article presents a case of a 60-year-old woman who was suspected of having a local recurrence on the chest wall following amputation of the left arm for a malignant mesenchymal tumor. The tumor did not show any invasion to adjacent structures thereby any necessity for chest wall resection and reconstruction. The patient underwent local excision of the tumor with reamputation of the branches of the lower brachial plexus, subclavian artery, and proximal one third of the clavicle. The tumor was eventually diagnosed as a brachial plexus neuroma. Although rare, cases that require left pneumonectomy with a transthoracic forequarter amputation including brachial plexus resection have been reported. Transection of the brachial plexus also may be needed during resection of a superior sulcus tumor. These cases often undergo an aggressive chest wall resection that includes the subclavian artery and vein in addition to the brachial plexus, particularly in tumors involving the lower truncus. Therefore, when recurrent chest wall tumor is suspected in such cases, stump neuroma resulting from transection of the brachial plexus should be included in the differential diagnosis. PMID- 19292196 TI - Lymphedema of the hand and forearm following fracture of the distal radius. AB - Lymphedema of the hand following a fracture of the distal radius is a disabling, but rare complication. Although the pathogenesis of this mechanism is poorly understood, extensive review of the literature suggests that an underlying cause of this lymphedema is psychogenic in nature. After numerous therapeutic modalities including intense physiotherapy, sympathetic nerve blocks and hospitalization, our patient still continued to persist with this debilitating disease. This article presents a case of Colles fracture complicated by nonpitting edema in a 62-year-old woman in whom psychogenic causes were not identified. The surgical procedure in our patient was uncomplicated and thus lymphedema was uncomplicated and thus lymphedema secondary to any vascular injury was ruled out. Questions that need to be addressed are whether the onset of the fracture induced an avascular anastomosis that led to the lymphedema. Our conclusions led us to believe the development of lymphedema of the distal radius following Colles fracture was idiopathic in our patient. PMID- 19292197 TI - Nailing before vascular repair in tibial fracture with associated arterial injury. AB - The incidence of arterial injuries in extremity fractures is approximately 1% to 2%. In patients with combined vascular and orthopedic injury but without a mangled extremity, the decision whether to give priority to the fracture fixation or to the vascular repair is still somewhat controversial. We successfully used the unreamed tibial nail before the arterial repair when treating an open tibial shaft fracture associated with a torn anterior tibial artery and present it as an illustrative case for the method. Fracture alignment was achieved manually and an unreamed tibial nail was introduced. The nail was secured only proximally by two interlocking screws inserted through jig. The procedure provided good alignment and length for end-to-end reconstruction of the anterior tibial artery. The distal interlocking screws were inserted at the end of vascular reconstruction. PMID- 19292198 TI - Post-traumatic cerebral fat embolism prior to operative repair of femoral and tibial fractures. AB - Rare cases of primary cerebral fat embolism as a postoperative complication have been reported. In addition, cerebral fat embolism occurring before operative repair without shunt lesion are more rarely reported. We report a patient with a posttraumatic cerebral fat embolism resulting in severe neurologic dysfunction without right to left shunt. A 25-year-old man was brought to the hospital immediately after a traffic accident. He sustained a right segmental femoral shaft fracture and a left tibial shaft fracture. The patient was alert with no neurological deficits. Approximately 13 hours after injury, the patient developed acute mental status deterioration and dyspnea. Magnetic resonance imaging of the brain revealed extensive multifocal infarction owing to embolic showering throughout nearly the entire brain parenchyma. Computed tomography with intravenous contrast revealed no evidence of embolism in the lung, abdomen, and pelvis. Transthoracic and transesophageal echocardiogram revealed no circulating embolic particles or intracardiac shunt. The patient underwent closed reduction and internal fixation with a reamed intramedullary nail in the tibia and underwent open reduction and internal fixation with a reamed nail in the femur at 5 days after injury. We performed decompression of a hematoma containing a large number of lipid droplets via a small incision in the femur shaft fracture, established suction drainage of the tibia medullar cavity, and applied a tourniquet to the thigh in the tibia shaft fracture. Supportive medical treatment included endotracheal ventilatory support and tracheostomy. The patient was discharged from the hospital 50 days after admission. On follow-up 2 months later, he had returned to activities of daily living, however a speech disturbance remained. PMID- 19292199 TI - Spinal cord compression due to vertebral hemangioma. AB - This article presents a case of multiple vertebral hemangiomas in a 58-year-old man with pain in the dorsal region and bilateral progressive foot numbness. Magnetic resonance imaging revealed multiple vertebral hemangiomas. One hemangioma at the T7 level demonstrated epidural extension, causing spinal cord compression. After treatment with radiotherapy, the patient's symptoms improved significantly. PMID- 19292200 TI - Postlaminectomy kyphosis in an achondroplastic adolescent treated for spinal stenosis. AB - We report a case of a skeletally immature achondroplastic adolescent with significant thoracolumbar lordosis who presented with neurogenic claudication and urinary incontinence progressing over a 1-year period. She underwent decompressive lumbar laminectomy from T12 to L5 with preservation of the facet joints. Over the ensuing 4 years of follow-up, she developed a progressive thoracolumbar kyphosis that progressed to 105 degrees. She remained neurologically intact and was fully ambulatory, but had severe back pain in the area of the deformity. Because of the severity and progression of her deformity, a combined anterior and posterior fusion and stabilization was required. We present this case and a review of the literature on spinal stenosis in achondroplasia and the complication of post-laminectomy kyphosis. PMID- 19292201 TI - Delayed presentation of metastatic melanoma of the cervical spine. AB - A 54-year-old woman presented with neck pain and bilateral shoulder pain. The pain was of varying intensity and was present with activity as well as at night and at rest. She had weakness of her triceps and finger intrinsic muscles. Her past medical history included stage IV melanoma for which she underwent an excision from the right flank area 25 years ago. She was cleared from any disease 5 years after the excision. Magnetic resonance imaging of the cervical spine revealed increased signal intensity in the C7 vertebral body with soft tissue extension into the epidural space. Two weeks after initial presentation the patient underwent a C7 corpectomy and fusion. The C7 vertebral body was grossly pathologic and tissue samples revealed metastatic malignant melanoma. During the 2 weeks following the fusion the patient developed increasing pain and generalized weakness. Repeat MRI of the cervical spine revealed a large soft tissue intensity extending in the spinal canal from C6 to T2. Several days later the patient developed an acute onset of lower extremity weakness as well as urinary incontinence. The patient opted to pursue hospice care, and she died 3 weeks later, 8 weeks after the initial presentation. PMID- 19292204 TI - Stabilization of pathologic long bone fractures with the Fixion expandable nail. AB - The Fixion intramedullary nail technology is safe and effective in the treatment of pathologic bone fractures. PMID- 19292207 TI - Effects of vascular bundle implantation on autograft, fresh-frozen allograft, and xenograft incorporation in a rabbit model. AB - This study examined the effects of vascular bundle implantation into a bone graft. Vascularized and nonvascularized autografts, allografts, and xenografts were placed inside defects in the proximal tibia in rabbits. Evaluation using radiographs, magnetic resonance imaging, bone scintigraphy, and microscopy showed autografts fused more rapidly than allografts and xenografts, and the majority of the vascularized grafts were incorporated completely. Autografts emerged as the gold standard. These findings indicate vessel implantation enhances and accelerates vascularization, new bone formation, and incorporation in autografts, allografts, and xenografts. PMID- 19292208 TI - The effect of postoperative immobilization on the healing of radiofrequency heat probe modified tissue: assessment of tissue length, stiffness, and morphology. AB - The effect of postoperative immobilization on the length, stiffness, and structure of connective tissue after radiofrequency probe shrinkage was examined in a rabbit patellar tendon model. Tendon lengths were measured before, immediately following, and 2, 4, and 8 weeks following heat application, with the contralateral tendon as a control. The animals were randomly assigned to three groups. In Group I, the controls, the animals were allowed free caged activity for 8 weeks. In Groups II and III, the experimental limb was immobilized at 15 degrees of knee flexion for 2 and 4 weeks, respectively, after which the animals were allowed 6 and 4 weeks of free caged activity. Changes in tendon length, stiffness, and cross-sectional area were compared using repeated measures ANOVA and differences between groups examined using Tukey's post-hoc analysis. Patellar tendon lengths were increased in all 3 groups at 8 weeks (P< .001). Tissue elongation was less extensive in group III compared to group I at 8 weeks (P< .001), while tissue cross-sectional area was decreased in both groups II and III compared to group I at 8 weeks (P< .001). All groups demonstrated decreases in tensile strength when compared to controls (P< .001). These findings suggest that postoperative immobilization is important following thermal shrinkage of connective tissue, as early activity can lead to stretching of the heat-modified tissue. PMID- 19292209 TI - Lumbar disk replacement with the ProDisc prosthesis. AB - Twenty-two ProDisc II prostheses (Spine Solutions, New York, New York) were implanted in 21 patients with degenerative disk disease at L5-S1 (19 disks) and L4-L5 (3 disks). After mean follow-up of 3.1 years (range, 17-49 months), pain intensity in all but 3 patients had improved from an average of 7.7 preoperatively to 4.6 postoperatively (P< .001) on a visual analog scale. Average Oswestry Disability Index score improved from 61 to 35 (P< .001). Radiographic reconstruction of the disk space height was achieved in all cases. Previous diskectomy at the implanted level and disk degeneration adjacent to previous fusion negatively influenced the results. PMID- 19292210 TI - Application of Hazard Analysis Critical Control Points to control surgical site infections in hip and knee arthroplasty. AB - This study examined critical points linked to the increase of surgical site infections in patients undergoing joint replacement. With this type of infection, it is not always possible to detect specific causes, and it often is necessary to perform a systematic review of assistance processes. Using Hazard Analysis Critical Control Points methodology, 4 stages of patients' pathway were identified: preoperative assessment, surgical procedure, postoperative assistance, and discharge. Surgical site infection risk factors were identified for each stage and categorized as patient conditions, staff and procedures, equipment, and environment. PMID- 19292211 TI - One-stage bilateral total joint arthroplasty: a prospective, comparative study of total hip and total knee replacement. AB - This prospective study compared the incidence of minor and major complications in 50 patients (100 joints) undergoing bilateral total knee arthroplasty (TKA) with 50 patients (100 joints) undergoing bilateral total hip arthroplasty (THA). Gender, comorbidities, American Society of Anesthesiologists scores, and body mass indices were similar in both groups. There was no difference in preoperative hemoglobin, operative time, anesthetic management, postoperative surveillance, and hemoglobin at discharge. However, the need for allogenic blood transfusion, despite preoperative autologous blood donation and retransfusion, was high for both groups (34% for bilateral TKA and 20% for bilateral THA. Bilateral THA patients had lower rates of total major and minor complications than bilateral TKA patients. Although cementless bilateral THA and cemented bilateral TKA can be performed efficiently and with relative safety in a select group of patients, the high rate of minor complications and in particular the need for allogenic transfusion in both groups is concerning and should be discussed with patients before surgery. PMID- 19292212 TI - Long-term results with the Ilizarov technique for tibiocalcaneal fusion. AB - Talar necrosis and infection are serious complications that have challenged orthopedic surgeons for years. In this study, 6 patients who underwent tibiocalcaneal fusion using the Ilizarov technique, predominantly for post traumatic talar osteitis, were reviewed after mean follow-up of 8 years. Solid fusion was obtained in all cases. In 5 patients, simultaneous lengthening was performed through a proximal tibial corticotomy. Complications were related primarily to pin insertion sites. The Ilizarov technique can be used successfully for tibiocalcaneal fusion in complex cases with talar osteitis that otherwise may ultimately require amputation. PMID- 19292213 TI - Comparison of distraction osteogenesis for congenital and acquired limb-length discrepancy in children. AB - This study evaluated 116 lower limb lengthenings in 88 consecutive patients (mean age, 13.5 years) treated for congenital (31 patients [46 lengthened segments]) and acquired (57 patients [70 lengthened segments]) limb-length discrepancy. Mean follow-up was 3.8 years. Mean lengthening index was 34+/-16 days/cm in the congenital group and 32+/-9 days/cm in the acquired group; this difference was not statistically significant (P> .05). Mean length of distraction regenerate was 6.4+/-3.7 cm in the congenital group and 5.7+/-2.9 cm in the acquired group; this difference was not statistically significant (P> .05). Lengthening percentage was significantly higher in the congenital group (25+/-17.6) than in the acquired group (19+/-15.4). The increase in the complications rate in congenital limb length discrepancy correlates with a significantly higher lengthening percentage. Bone regenerate is produced equally regardless of whether the etiology is acquired or congenital. PMID- 19292214 TI - Comparison of two tension-band fixation materials and techniques in transverse patella fractures: a biomechanical study. AB - This study compared the biomechanical properties of 2 tension-band techniques with stainless steel wire and ultra high molecular weight polyethylene (UHMWPE) cable in a patella fracture model. Transverse patella fractures were simulated in 8 cadaver knees and fixated with figure-of-8 and parallel wire configurations in combination with Kirschner wires. Identical configurations were tested with UHMWPE cable. Specimens were mounted to a testing apparatus and the quadriceps was used to extend the knees from 90 degrees to 0 degrees; 4 knees were tested under monotonic loading, and 4 knees were tested under cyclic loading. Under monotonic loading, average fracture gap was 0.50 and 0.57 mm for steel wire and UHMWPE cable, respectively, in the figure-of-8 construct compared with 0.16 and 0.04 mm, respectively, in the parallel wire construct. Under cyclic loading, average fracture gap was 1.45 and 1.66 mm for steel wire and UHMWPE cable, respectively, in the figure-of-8 construct compared with 0.45 and 0.60 mm, respectively, in the parallel wire construct. A statistically significant effect of technique was found, with the parallel wire construct performing better than the figure-of-8 construct in both loading models. There was no effect of material or interaction. In this biomechanical model, parallel wires performed better than the figure-of-8 configuration in both loading regimens, and UHMWPE cable performed similarly to 18-gauge steel wire. PMID- 19292215 TI - Comparison of screw and screwless fixation in cementless total knee arthroplasty. AB - This study compared two groups of consecutive patients undergoing total knee arthroplasty (TKA) with an identical cementless tibial baseplate supplementally fixated with and without screws. In 58 TKAs, two 6.5-mm cancellous screws were used for fixation, and in 58 TKAs, screwless fixation was used. Clinical evaluation was performed using the Hospital for Special Surgery knee score, and fluoroscopically guided radiographs were evaluated for radiolucencies. In the screw fixation group, average follow-up was 82+/-6 months (range, 59-89 months) and average patient age at surgery was 63 years. For the screwless fixation group, average follow-up was 67+/-5 months (range, 48-76 months) and average patient age at surgery was 62 years. Average knee score was 97 for the screw fixation group and 98 for the screwless fixation group. This study demonstrated stability in cementless TKA both with and without screw fixation in the mid-term using the Natural-Knee II tibial component. PMID- 19292216 TI - Comparison of autograft and allograft fixation in Pemberton osteotomy. AB - This retrospective study compared the results of autograft and allograft fixation in 29 children (36 hips) who underwent Pemberton osteotomy. Autograft fixation was used in 21 hips and allograft fixation was used in 15 hips. In the autograft group, 76% of patients had a successful result; 4 of 8 of patients with a neuromuscular disorder in this group had a successful result. In the allograft group, 93% of patients had a successful result; 6 of 7 of patients with a neuromuscular disorder in this group had a successful result. Pemberton osteotomy performed with allograft fixation provided similar or better results than autograft, especially in children with neuromuscular disorders. LEVEL OF EVIDENCE: diagnostic study, Level III-2 (retrospective cohort study). PMID- 19292217 TI - Treatment of intertrochanteric nonunion of the proximal femur using the S-ROM prosthesis. AB - Nonunion is an uncommon yet devastating complication following intertrochanteric hip fracture, and few reports in the literature address this problem. This study examines the results of intertrochanteric nonunion treated with the uncemented S ROM prosthesis in 10 patients. All patients demonstrated clinically significant pain relief and return to ambulation following hemiarthroplasty or total hip arthroplasty. Average Harris hip score was 86 after an average follow-up of 30 months; there was a trend toward better results in cases of pure intertrochanteric fractures compared to cases with subtrochanteric involvement. Radiographic follow-up demonstrated stable ingrowth in 9 cases, and few complications occurred in this series. PMID- 19292218 TI - Mentoring healers. PMID- 19292221 TI - Acquired boneless forearm as a complication of traditional bone setting. AB - An 8-year-old girl sustained closed fracture of the right ulna 10 weeks prior to presentation. She was taken to a traditional bone setter who applied a tight splint. The patient reported pain, but the splint was not removed. A week after application of the splint, a foul odor was detected and removal of the splint showed extensive exposure of the forearm bones. Above elbow amputation was rejected by the patient's parents when she was taken to hospital, where she was admitted for 8 weeks by a second traditional bone setter. A trained nurse applied herbal concoctions and dressed the wound daily in anticipation that the skin would cover the exposed bone fragments. She was brought to our hospital for wound dressing so that the skin would cover the exposed bones fragments. Examination revealed a grossly shortened right forearm--by 7 cm compared with her left- extensive exposure of both radius and ulna at the anterior aspect of the forearm, and loss of sensation and movement of the fingers. Radiographs showed sequestrated radius and ulna with involucrum around the olecranon process. Above elbow amputation was offered to the patient but the parents again declined. The forearm bones detached while scrubbing the wound for review and removal of the sequestrated bone. The wound healed within one and a half weeks of dressing, resulting in an acquired boneless forearm. PMID- 19292222 TI - Spondylolysis of the second lumbar vertebra treated with segmental wiring and bone grafting. AB - Spondylolysis affects mostly the lower lumbar spine and rarely the upper lumbar spine. In a literature research, we found that the descriptions of spondylolysis of the upper lumbar spine had been reported mainly with the outcomes of conservative treatment using lumbosacral supports. However, an indication of surgical treatment has rarely been reported. Ravichandran et al reported 2 cases of spinal fusion, decompression or a combination of these procedures, but the procedures have not proved satisfactory in cases of upper lumbar spondylolysis. We found no reports of segmental wire fixation and bone grafting for upper lumbar spondylolysis. Herein, we report spondylolysis of the second lumbar vertebra in a 27-year-old man. He presented with pain and tenderness at the L2 spinous process, and swollen paravertebral muscles. The patient first became aware of lumbago at age 24 years. Much of his work involved heavy labor. He was diagnosed with spondylolysis of L2 and treated conservatively by a general orthopedist from age 25 to 27 years, but without improvement. We performed segmental wire fixation of the transverse and spinous processes of L2, followed by a bone graft. Six months after surgery, the lumbago had resolved and the patient was able to return to work. Three years after surgery, his pain is completely resolved. This is the first report in the English literature in which a successful indication for symptomatic spondylolysis in the upper lumbar spine is described. PMID- 19292223 TI - Florid reactive periostitis ossificans of the distal ulna. AB - Florid reactive periostitis is the term used to describe a benign disease characterized by aggressive periosteal reaction and benign soft-tissue inflammation. Although it is considered rare, there are numerous reports in the literature that refer to this disease with different names such as parosteal fasciitis, fasciitis ossificans, benign fibro-osseous pseudotumor, pseudomalignant osseous tumor of soft tissue, and bizarre parosteal osteochondromatous proliferation. As a result, the nomenclature is confusing, and some authors have placed florid reactive periostitis ossificans into the heterogeneous group of pseudomalignant osseous tumors of soft tissue or proliferative periosteal processes, whereas other authors place this entity in the myositis ossificans group. In the same manner, florid reactive periostitis has been considered to be a previous stage of bizarre parosteal osteochondromatous proliferations. This article presents a case of florid reactive periostitis ossificans of the distal ulna in a 13-year-old boy. The patient presented with a painful lesion in the distal ulna, and plain radiographs suggested the presence of a quickly growing periosteal lesion with associated calcification and soft tissue mass. Histologically, the appearance was that of reactive periostitis. The clinical, radiological, and histologic features of florid reactive periostitis are described. PMID- 19292224 TI - Treatment of avulsion fractures of the second metacarpal base with a miniplate and screws. PMID- 19292225 TI - Progressive Gorham disease of the forearm. AB - Gorham's-Stout disease is a rare but potentially debilitating disease consisting of massive bone osteolysis and bone resorption associated with vascular proliferation and increased osteoclastic activity. Although it can present in a wide variety of forms, it typically involves bones formed by intramembranous ossification such as the skull, pelvis, and scapula. It can occur spontaneously or after trauma. Most cases are monofocal and resolved spontaneously, although there are reports of multifocal and rapidly progressing disease. It typically presents as disuse muscle atrophy or pathologic fracture during the second through fourth decades of life, yet it has also been reported in childhood and in the elderly. The etiology of Gorham's disease remains to be fully elucidated. Gorham attributed the origin of the disease to uncontrolled proliferation of small vessels eating away bone tissue. Other authors attribute the cause of the disease to increased osteoclastic activity mediated by elevated cytokine levels and increased osteoclastic differentiation. Treatment is not established and focuses at stopping osteoclastic activity and angiogenic proliferation. Radiation therapy, chemotherapy, bone grafting, and antiresorptives medications have all been used with different degrees of success. In an effort to further characterize this elusive disease, we report on an unusual presentation of a patient with Gorham's disease of the radius spreading to the ulna and then the proximal humerus with a 13-year follow-up. To our knowledge this is the first report in the literature of a saltatory type of Gorham's disease spreading from bone to bone across a joint. PMID- 19292226 TI - Acute fracture of the acetabulum secondary to a convulsive seizure 3 years after total hip arthroplasty. AB - While aseptic loosening, osteolysis, and infection are the most common causes of failure after total hip arthroplasty (THA), late hip pain can also be the result of acetabular fracture related to trauma and resultant prosthetic failure. However, atraumatic fracture of the acetabulum around a well-fixed acetabular component is unusual. We present a patient with an acetabular fracture resulting from a generalized convulsive attack 3 years after an uncomplicated primary THA. A 33-year-old man presented with acute left hip pain. He had chronic renal insufficiency and had undergone bilateral THA due to avascular necrosis. The night prior to his admission, he suffered a generalized convulsive attack with severe extremity contractions. Afterwards, he had acute left groin pain and had difficulty walking. Physical examination revealed moderate left hip pain as well as a 1-cm shortening of the affected limb. Radiological examination demonstrated an acetabular fracture with medial wall comminution. The acetabular component had migrated medially and rotated horizontally. Revision of the acetabular component with a reinforcement ring and implantation of a cemented acetabular component was realized. Severe muscle spasms during generalized seizures are known to lead to various musculoskeletal injuries (fractures of the proximal humerus, femur, acetabulum, and dislocation of the shoulder). Seizures could also lead to acute periprosthetic fracture of the acetabulum in patients with osteopenia. Therefore careful reaming is required to avoid overmedialization of the acetabular component in those patients. PMID- 19292227 TI - Unusual radiologic presentation of monostotic fibrous dysplasia. AB - Fibrous dysplasia is a benign fibro-osseous lesion that can occur as an isolated skeletal lesion (monostotic form) or affect multiple skeletal sites (polyostotic form). In addition, fibrous dysplasia may be associated with single or multiple endocrinopathies, or with precocious puberty and cutaneous hyperpigmentation in McCune-Albright syndrome. Fibrous dysplasia usually appears as a well-defined radiolucent medullary lesion that is irregular, mildly expansive, and with a hazy opacity classically described as "ground glass" in appearance. In the long tubular bones, fibrous dysplasia may cause expansion of the bone contour with cortical thinning and endosteal scalloping. The shaft is typically involved, but the metaphysis may also be affected. These changes are usually recognizable on plain radiographs, but in cases where the lesion is difficult to visualize computed tomography may be beneficial. However, fibrous dysplasia may present with radiographic features that may mimic other benign fibro-osseous lesions, may be associated with other lesions, and even be confused with certain types of malignancies. We present a case of monostotic fibrous dysplasia in the proximal tibia of an 11-year-old girl with the unusual radiologic features of multifocal lytic lesions and cortical disruption. While most often the clinical presentation, laboratory findings, and imaging studies permit the diagnosis of fibrous dysplasia, in certain cases this may pose a diagnostic challenge. The diagnosis was confirmed by open biopsy. PMID- 19292228 TI - Intraosseous schwannoma of the frontal bone. AB - Schwannomas or neurilemomas are relatively frequent benign tumors; however, intraosseous schwannomas are rare. The most common site for intraosseous schwannoma is the mandible. Other sites include the vertebra, ulna, humerus, femur, tibia, patella, scapula, rib, and small bones of the hands. The skull vault is a rare site for intraosseous schwannoma, and only 4 cases have been reported previously in the literature. This article presents a case of intraosseous schwannoma in the frontal bone in an 11-year-old boy. Radiologically, intraosseous schwannomas are lytic, sharply demarcated defects that often expand the involved bone. Perilesional sclerosis may surround the area of central lucency. Histologic features of intraosseous schwannomas are similar to that observed for soft-tissue schwannomas including Antoni type A areas (compactly arranged spindle-shaped cells with palisading or Verocay bodies) and Antoni type B areas (widely separated cells with loose intervening collagenous matrix). In our patient, typical radiologic and histologic findings including immunohistochemistry and electron microscopy results led to a diagnosis of intraosseous schwannoma. PMID- 19292229 TI - Adult onset flatfoot associated with an intramuscular hemangioma of the posterior tibialis muscle. AB - A 30-year-old man was referred for a painful mass in the left calf and planovalgus deformity of the left foot. The pain had been present for 15 years but recently had become severe. On examination there was a visible, exquisitely tender mass in the medial distal aspect of the lower leg. There was valgus deformity of the hindfoot, abduction of the forefoot, and complete loss of the longitudinal arch which was passively correctable to neutral. Magnetic resonance imaging showed a 3.035.5312.0-cm complex mass in the calf with a "bag of worms" appearance consistent with a vascular tumor. An incisional biopsy resulted in diagnosis of benign intermuscular hemangioma. A marginal resection of the mass was later performed. During operative resection, the tumor was found to involve the entire posterior tibialis muscle to the musculotendinous junction. The patient began full weight bearing 4 weeks postoperatively and was placed in an ankle-foot orthosis for correction of the planovalgus. At 19 months postoperative, he reported no pain and mild weakness of the left foot, and he had returned to full-time work. There was no recurrence of the tumor. Intermuscular hemangioma of the calf may cause acquired spasticity and equinus deformity of the foot. In addition, intramuscular hemangiomas have been reported to cause muscle weakness and tendon rupture. In this patient, the authors believe that the posterior tibialis muscle was weakened and elongated secondary to the replacement of the muscle tissue by the tumor, ultimately leading to the planovalgus deformity. PMID- 19292230 TI - Multidisciplinary investigation in Down syndrome: bear in mind. AB - Patients who suffer from Down Syndrome may have a number of orthopedic and rheumatologic disorders. Recent life-table study of live-born individuals with Down Syndrome reports a survival rate of >50% to the age of 50. Therefore some osteoporotic-related disorders have to be considered. Femoral neck fracture is a typical disease when these patients are elderly, often due to traumatic episodes or to violent seizures. We report the case of a 50-year-old Down Syndrome patient who had progressive loss of mobility and standing capacity associated with mental confusion, first considered to be a neurological disturbance. He lived with his family and led a standard lifestyle for his condition with a normal ability to walk. The parents reported he had not fallen and that, before the episode, he had been able to walk and stand normally. After examination by a multidisciplinary team of physicians, the diagnosis was a bilateral femoral neck fracture. The patient was then treated with a one-stage bilateral hip hemiarthroplasty. The pathogenesis of osteoporosis Down Syndrome patients is multifactorial and includes a sedentary lifestyle and poor mobility, endocrine abnormalities, and anticonvulsant medications. The musculoskeletal disorders, however, are not the only medical problems these patients have to face. Because of the limited communicative skills, clinical symptoms and signs of early disease frequently go undetected or are misdiagnosed for long periods of time. For this reason we recommend annual systematic screening for health problems including screening for osteoporosis. PMID- 19292231 TI - Vanishing bone disease: review and case reports. AB - Vanishing bone disease is a rare idiopathic disease, leading to extensive loss of bony matrix, replaced by proliferating thin-walled vascular channels and fibrous tissue. There are >191 cases reported in the English literature. Gorham and Stout made the first overview of the disease in 1955 and they first presented 24 cases known at that time. The etiology remains speculative, the prognosis unpredictable, and effective therapy still unknown. The disease can be monostotic or polyostotic although multicentric involvement is exceptional. We report 2 cases of a histologically studied vanishing bone disease involving the humerus and the femoral head. The patients' past history was noncontributory. The radiographic study revealed a destructive lesion of the left humerus in the first case, and complete disappearance of the femoral head in the second case. Laboratory findings including hormonological tests revealed no evidence of metabolic, immunologic, neoplastic, or infection etiology. Histopathological findings of the 2 cases revealed thickened bone of lamellar structure without marrow cavities next to fibrous tissue, with few fibroblasts and a small number of newly formed vascular channels. The prognosis varies from slight disability to death by involvement of vital skeletal structures. The treatment of vanishing bone disease is controversial. Several treatment modalities have been proposed. Surgical intervention has been suggested as a method of choice by many authors and concerns local resection of the affected bone, with or without replacement prosthesis or bone grafts. PMID- 19292232 TI - Histologic analysis of allograft mixed with hydroxyapatite-tricalcium phosphate used in revision femoral impaction bone grafting. AB - Failed total hip arthroplasty (THA) often is accompanied by loss of bone stock. In cases in which revision THA becomes necessary, adequate stabilization of the implant can be difficult because of the bone loss. Bone can be reconstructed by using tightly packed morselized cancellous bone graft in combination with a cemented stem. However, worldwide availability of human bone is limited, and risks such as infection and antigenicity are associated with the use of allogeneic bone. These risks have led to the development of synthetic bone graft extenders. The objective of impaction bone grafting is to achieve stability of an implant and subsequently allow the restoration of living bone stock. To reduce the amount of allograft consumed, a mixture of morselized allograft and hydroxyapatite-tricalcium phosphate was used for impaction grafting in a series of 15 patients. In 2 patients who required re-revision arthroplasty, the areas of impacted graft were analyzed histologically. The results showed a complex mixture of residual hydroxyapatite-tricalcium phosphate, necrotic bone (graft), viable bone, and fibrous tissue. Although several pre-clinical studies have described the expansion of allograft with synthetic materials, this article represents the first description of histologic findings in human femoral impaction grafting with a combination of allograft and hydroxyapatite-tricalcium phosphate. PMID- 19292233 TI - Cervical kyphosis and thoracic lordoscoliosis in a patient with cerebral palsy. AB - The management of cerebral palsy and spinal deformity can be challenging for spine surgeons. Scoliosis is the most common spinal deformity associated with cerebral palsy; however, the development of thoracic lordoscoliosis is rare. Cervical kyphosis in cerebral palsy patients is associated primarily with athetoid cerebral palsy. This article presents a case of progressive myelopathy and concurrent cervical kyphosis and thoracic lordoscoliosis in a 17-year-old spastic quadriplegic cerebral palsy patient who underwent 2-stage treatment. In the first stage, the cervical spine was managed with a 2-level cervical corpectomy of C5 and C6, with multilevel laminectomies, lateral mass plating, and wire fixation. In the second stage 3 months later, the thoracic deformity was corrected through a combined anterior spinal release of the thoracic spine with posterior spinal segmental instrumentation and sublaminar wiring. Postoperatively, the patient successfully returned to independent ambulation. PMID- 19292234 TI - Failure of constrained acetabular liner without metal ring disruption. AB - Constrained acetabular liners can fail leading to recurrent dislocation. Failure can occur at any of the five possible interfaces: bone-acetabular shell, acetabular shell-constrained liner insert, constrained liner insert-bipolar head, bipolar head- femoral head and femoral head-trunion. We report a patient who presented with dissociation of the cemented Osteonics acetabular constrained liner (Stryker-Howmedica-Osteonics, Rutherford, New Jersey). The failure interface was at the factory pre-assembled constrained liner insert-bipolar head without any locking ring failure; instead there was deformation of the constrained liner insert's polyethylene rim, which facilitated dissociation. To our knowledge, there are no previous reports of such a failure mode pertaining to this type of constrained liner. Constrained acetabular liners are indicated during primary or revision total hip arthroplasty for patients who are at high risk for dislocations or who have had recurrent dislocations. Failure rates (typically recurrent dislocation) range from 4% to 29% at mid-term follow-up. The first report on the Osteonics acetabular constrained liner was published in 1994. Failures have been reported previously to occur at surgically controllable interfaces, such as the acetabular shell from the bony surface and the constrained liner insert from the acetabular metal shell, and have been attributable to excessive constraint or improper technique. All dissociations pertaining to factory-preassembled component interfaces have been attributed to breakage of the locking ring. This article presents the first case of disengagement of the tripolar constrained liner without disruption of the locking ring. PMID- 19292235 TI - A rare trifocal presentation of Clostridium septicum myonecrosis. AB - Gas gangrene or clostridial myonecrosis is a rare, life threatening infection of the muscle that is typically associated with recent surgery or trauma. It usually affects older individuals who suffer from vascular insufficiency or who are immunocompromised. The typical pathogen is Clostridium perfringens. Atraumatic gas gangrene caused by Clostridium septicum is less common and has a unique association with colon cancer. This condition aggressively spreads through the muscular tissue and often culminates in septic shock. Fatality occurs within 48 hours if left untreated yet, despite emergent treatment the mortality rate is >50% without any comorbidities. Given the propensity to occur in the extremities the orthopedic surgeon must be familiar with its presentation, diagnosis, and treatment. This article presents a case of a 16-year-old girl who presented to the emergency room with an 18-hour history of malaise and progressive right arm discomfort and swelling. The identification and emergent treatment of necrotizing soft tissue infections is important and often falls on the orthopedic surgeon, especially when isolated to the extremities. Subtle initial presentation and rapid progression of the infection are classic findings of necrotizing soft tissue infections, particularly with clostridial myonecrosis and necrotizing fasciitis. Clostridium myonecrosis is a rare orthopedic condition with high mortality. Early diagnosis and debridement is imperative. A team approach to managing these patients postoperatively is necessary. The use of hyperbaric oxygen can be considered, however, it may not be readily available and its use is unproven in human studies. Due to its high mortality, one should err on the side of more aggressive resective/ablative surgery. PMID- 19292236 TI - Treatment of posterior sternoclavicular dislocation with locking plate osteosynthesis. AB - Posterior dislocations of the sternoclavicular joint are rare injuries, representing approximately 3% of all major insults to the shoulder girdle. Despite their relative infrequency, they must be diagnosed and treated early because of the proximity of the sternoclavicular joint to sensitive anatomic structures in the superior mediastinum. Physical examination can yield limited findings, and a high index of suspicion is warranted for this injury pattern especially with impingement signs and symptoms: dyspnea, dysphagia, dysphonia, brachial plexus injury, and vascular alterations. Evaluation with plain anteroposterior radiographs is difficult because of superimposed ribs and lungs. A computed tomography scan is required for appropriate evaluation of this injury pattern. Once posterior dislocation of the sternoclavicular joint has been diagnosed, closed reduction should be attempted as soon as possible with a thoracic surgeon available. Most posterior sternoclavicular dislocations can be treated successfully with closed reduction and 6 to 8 weeks of immobilization. Recurrent dislocations or chronic dislocations require open treatment for stabilization. Precisely which technique should be used to obtain stable internal fixation is controversial, but ligament repair with reconstruction appears to be the most widely accepted. This article reports two such cases of recurrent dislocation that were treated with locking plate osteosynthesis as opposed to more commonly used soft tissue stabilization procedures. Both patients experienced a nearly full return to function and are currently doing well. A review of the current literature regarding this subject matter is also included. PMID- 19292237 TI - Acute isolated lateral leg compartment syndrome following a peroneus longus muscle tear. AB - Acute compartment syndrome is a rare orthopedic emergency caused by orthopedic, vascular, iatrogenic, or soft tissue injury that elevates the pressure within a closed fascial space. Untreated acute compartment syndrome leads to ischemia of the muscles and nerves within the compartment with a resultant loss of motor and sensory function, death of skeletal muscle, and subsequent contracture and deformity. This article presents a case of compartment syndrome in a 33-year-old man following a peroneus longus muscle tear. The patient underwent emergent fasciotomy and decompression of the lateral compartment, and a hematoma located at the musculotendonous junction of the peroneus longus was evacuated. The patient was placed in a posterior splint with the ankle at 90 degrees and elevated. Postoperatively, the patient reported complete resolution of pain, numbness, and tingling. The patient underwent delayed skin closure on the second postoperative day and subsequently had an uneventful recovery. By 4 months postoperatively, he had resumed full activity. Knowledge of the signs and symptoms of compartment syndrome and a high index of suspicion is paramount as this injury can occur after a seemingly innocuous injury such as an ankle inversion injury. PMID- 19292238 TI - Femoral neck fracture as an atypical presentation of child abuse. AB - Femoral neck fractures account for 46% of the proximal femur fractures in children; however, the overall incidence of proximal femur fractures in children is <1%. Pathologic causes for proximal femur fractures include local or global causes of bone weakness, such as metabolic bone conditions or neoplastic causes. Nonpathologic causes of femur fractures are caused almost exclusively by high energy trauma and are readily identified. Nonaccidental injury leading to femoral neck fracture is unusual and can be challenging to diagnose when the medical history is not forthcoming. Femoral neck fractures in children usually are a consequence of high-energy trauma or less frequently are associated with pathologic conditions. If neither condition is readily identified, the possibility of nonaccidental injury should be considered. This article presents a case of nonaccidental injury with an atypical presentation in a 3-year-old girl with a femoral neck fracture and discusses the diagnostic rationale for a diagnosis of nonaccidental injury. This case report highlights the magnitude of child abuse and its early recognition in the orthopedic community. PMID- 19292246 TI - Comparison of ligament reconstruction tendon interposition and trapeziometacarpal interposition arthroplasty for basal joint arthritis. AB - This study compared patients with basal joint arthritis who underwent either ligament reconstruction tendon interposition (LRTI) or trapeziometacarpal interposition arthroplasty (TMIA). Twenty-two consecutive LRTI and 22 TMIA procedures were compared. Arthritis was graded using the staging system of Eaton. In the LRTI group, 1 patient (4.5%) had stage II, 3 patients (13.6%) had stage III, and 14 patients (15 LRTI procedures; 68.1%) had stage IV disease; 3 patients had previous silicone implants (13.6%). In the TMIA group, 3 patients (13.6%) had stage II and 19 patients (86.4%) had stage III disease. Mean age was 62.5 years in LRTI patients and 54.7 years in TMIA patients (P=.005). Mean follow-up was 48 months for both groups. Thumb shortening was determined using preoperative lateral and follow-up pinch lateral radiographs. Functional outcomes and patient satisfaction were analyzed. Pinch strength averaged 8.1 lb for LRTI and 12.6 lbs for TMIA patients; this difference was significant (P<.002). Patients in both groups had high overall satisfaction rates, with scores averaging 8.5 for LRTI patients and 9.2 for TMIA patients. Thumb metacarpal proximal migration was 20.5% in the LRTI group, and 6.5% in the TMIA group; this difference was significant (P=.0006). There was no statistically significant linear correlation between proximal migration and pinch strength (R=0.32, P=.13). Five TMIA patients required revision to LRTI. Patients in both groups reported high satisfaction. Pinch strength was greater in TMIA patients, and proximal migration of the thumb metacarpal was greater in LRTI patients. There was no correlation between proximal metacarpal migration and pinch strength. Increased pinch strength with TMIA may not be sufficiently advantageous to outweigh the risk for revision surgery. PMID- 19292247 TI - The effect of collar on aseptic loosening and proximal femoral bone resorption in hybrid total hip arthroplasty. AB - This study compared proximal femoral bone resorption in hybrid total hip arthroplasty cases that had poor or good contact between the collar and proximal medial femoral neck. A total of 94 patients (102 hips) comprised the study group. Mean patient age was 52 years, and mean follow-up was 4.86 years. Bone resorption of the proximal femur was evaluated with immediate postoperative and follow-up anteroposterior and lateral radiographs. Statistical analysis using the Mann Whitney test showed no significant difference between the groups that had good or poor contact. The collar did not prevent calcar resorption even when ideal contact was achieved. PMID- 19292248 TI - Classification of anterosuperior glenoid labrum variants and their association with shoulder pathology. AB - This prospective study of 367 consecutive shoulder arthroscopies assessed variants of the anterosuperior glenoid labrum and associated shoulder pathology. Thirty-three shoulders were excluded because of prior surgery, septic arthritis, or adhesive capsulitis. Anterosuperior glenoid variants were classified as: type I, cordlike middle glenohumeral ligament without sublabral foramen; type II, sublabral foramen without a cordlike middle glenohumeral ligament; type III, sublabral foramen with a cordlike middle glenohumeral ligament; and type IV, absent anterosuperior labrum with the anterior aspect of the superior labrum continuous with a cordlike middle glenohumeral ligament. The presence of these variants was correlated with the incidence of shoulder pathology found on arthroscopic inspection. Of 334 shoulders, 118 (35.3%) had variants of the anterosuperior glenoid labrum. Of these, 32 (27.1%) were type I, 27 (22.9%) were type II, 34 (28.8%) were type III, and 25 (21.2%) were type IV. The incidence of advanced superior labrum anterior-posterior lesions in the 86 shoulders displaying a type II, III, or IV variant was significantly higher than in shoulders with no anterosuperior variant (48.8% versus 23.6%, P<.001). Other pathologic findings were not significantly increased in shoulders with variants compared to those without. LEVEL OF EVIDENCE: Level 1. PMID- 19292249 TI - Management of falls after total knee arthroplasty. AB - This study evaluated 78 patients who fell after total knee arthroplasty. Eight patients (10.3%) had soft-tissue injuries that led to a poor clinical course (deep infection, unplanned return to the operating room, temporary or permanent resection arthroplasty, or knee arthrodesis). Factors related to a poor clinical course included copious bleeding from the surgical wound immediately after falling, falling within 8 weeks of arthroplasty, and rupturing the parapatellar surgical wound and quadriceps repair. Patients who fall and have bleeding from their surgical wound should receive emergency surgical irrigation and debridement. A treatment protocol for patients who fall after total knee arthroplasty is presented. PMID- 19292250 TI - The effect of cup placement in cementless total hip arthroplasty on the wear rate of polyethylene. AB - There are many reports on cup placement, inclination, anteversion, and position. This article strictly defines the area of cup placement based on data from 185 healthy hip centers and Ranawat's triangle. Linear wear rate (LWR) was measured in 55 total hip arthroplasty (THA) cases and categorized as low or high. The relationships between these categories and cup size, inclination, anteversion, position, age, follow-up period, bone graft, cup osteolysis, and stem osteolysis were investigated. PMID- 19292251 TI - Immediate weight bearing after cementless total hip arthroplasty. AB - This study analyzed 399 Balance femoral stems to determine whether immediate full weight bearing following total hip arthroplasty affected radiographic osseointegration. Postoperatively, all patients were permitted unlimited weight bearing. Findings demonstrated osseointegration was successful in 99.5% of femoral components. Of these, 0.5% subsided in the first 6 postoperative weeks and then osseointegrated; 0.5% failed to osseointegrate. Patients with Dorr C bone had a significantly increased rate of subsidence (P=.006). These findings indicate the Balance stem reliably osseointegrates without subsidence in patients with Dorr A or B bone, despite immediate full weight bearing. PMID- 19292252 TI - Microporous polysaccharide hemospheres do not inhibit bone healing compared to bone wax or microfibrillar collagen. AB - Topical hemostats may reduce bone bleeding but the presence of residual matter may inhibit bone growth. We compared a recently approved hemostat, microporous polysaccharide hemospheres (Arista AH, Medafor Inc, Minneapolis, Minnesota), with conventional hemostatic agents in a rabbit calvarial model. Standard defects were created and microfibrillar collagen (Avitene; C.R. Bard Inc, Murray Hill, New Jersey), bonewax, or microporous polysaccharide hemospheres was applied. Bone growth was evaluated after fluorescent labeling the mineralization front. At 7 weeks bonewax and microfibrillar collagen had reduced bone growth compared to control or microporous polysaccharide hemospheres. Microporous polysaccharide hemospheres and control animals also had reduced rabbit calvarial defects compared to bonewax or microfibrillar collagen. PMID- 19292253 TI - Bacterial growth on articulating spacers: an in vitro study. AB - We fashioned cement disk-shaped spacer models using antibiotic-loaded Palacos and embedded polyethylene and titanium into the surface of half of the models and inoculated the models with methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant S aureus (MRSA), or Staphylococcus epidermidis, and placed them in nutrient broth. Vancomycin was loaded into the cement of the MRSA spacer models and tobramycin into the MSSA and Staphylococcus epidermidis models. In the MSSA and MRSA models, no organisms survived beyond 48 hours in the antibiotic bath regardless of the presence of additional materials. At 96 hours, 86.6% of models with only antibiotic cement had viable Staphylococcus epidermidis, while 80% of models with antibiotic cement, polyethylene, and titanium had viable Staphylococcus epidermidis. Adding polyethylene and titanium to antibiotic-loaded cement does not promote bacterial survival. PMID- 19292254 TI - S-ROM hip prosthesis: 10- to 14-year results. AB - This retrospective study evaluated the results of 55 S-ROM femoral components (45 patients) at 10 to 14 years postoperatively. Twelve patients (15 hips) were lost to follow-up. Survivorship of the prosthesis was determined in the remaining 33 patients (40 hips). Harris hip scores and radiographic analyses were performed. Thirty-four hips were performed in a primary fashion and 6 for revision arthroplasty. There was 100% survival for aseptic loosening in the primary arthroplasty group and 1 failure in the revision group. This study shows excellent long-term results of the S-ROM prosthesis at 10 to 14 years. PMID- 19292255 TI - Fixation strength of tibial inlay graft with one versus two screws: a cadaveric study. AB - Various modes of fixation have been advocated for posterior cruciate ligament reconstruction. This study compared pullout forces and modes of failure for 2 commonly used techniques. Ten Achilles tendon-bone plug grafts were fixed to tibial troughs using either one 6.5-mm or two 4-mm cancellous screws positioned anatomically and tested to failure. The constructs did not differ statistically in mean pullout forces (385.2 and 358.8 N for the 6.5-mm and 4-mm screw constructs, respectively). The 2-screw construct failed by pullout, whereas the single-screw fixation failed through fracture. This study provides insight into the fixation used for posterior cruciate ligament reconstruction. PMID- 19292256 TI - Surgical treatment of severe thoracic scoliosis in skeletally mature patients. AB - The few number of severe thoracic scoliosis requiring surgical treatment makes the description of its therapeutic course difficult. Twenty-one cases of severe thoracic scoliosis with Cobb's angle >90 degrees were treated surgically in the past 20 years. Surgical treatment was performed because of an evolutive scoliosis or in case of respiratory or functional impairment. All patients underwent posterior approach after a preoperative preparation by the use of halo-traction. In 6 cases, a previous surgical posterior spinal release was performed before the halo-traction period. Fusion achieved in all cases and the final correction was generally better than preoperative reducibility. The surgical course was guided by the poor functional status of these patients, especially concerning respiratory function. Spinal instrumentation was made up of 3 rods with few spinal implants in the majority of the cases. Surgical correction was made by means of rods narrowing without any rod's rotation procedures. Functional improvement, especially respiratory was noted in all the cases. Nevertheless, postoperative spirometries were not significantly different from the preoperative ones. The aesthetic improvement of the rib hump was disappointing making necessary a complementary thoracoplasty in two cases. Surgical treatment of severe thoracic scoliosis remains justified in adults because of a progressive functional and radiological deterioration with respiratory compromise. We performed spinal fusion by posterior approach only, after a preoperative period of halo-traction. The final correction depends on the preoperative reducibility. We noted a functional and respiratory improvement in all the cases. PMID- 19292257 TI - Reconstruction of the medial patellofemoral ligament using a quadriceps tendon graft: a case series. AB - This study assessed medial patellofemoral ligament reconstruction using a partial thickness quadriceps tendon graft in 14 knees in 13 patients. Nine patients were available for follow-up after a minimum of 24 months. Final patient evaluation was performed an average of 42 months postoperatively (range, 28-65 months). Primary outcome measure was occurrence of patellar dislocation postoperatively, and outcomes were quantified using the Kujala questionnaire and Crosby and Insall criteria. Postoperatively, no patient reported patellar dislocation. Using Crosby and Insall criteria, good or excellent results were achieved in 100% of patients. Mean Kujala score was 91.9. Postoperatively, all patients reported their knee was improved. These findings indicate medial patellofemoral ligament reconstruction with a quadriceps tendon graft is effective in preventing patellar dislocation and improving quality of life. However, patients should be counseled this procedure is indicated primarily for the treatment of recurrent instability, and postoperative relief of anterior knee pain is inconsistent. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 19292258 TI - Articular cartilage thickness of the humeral head: an anatomic study. AB - This study determined the thickness of normal humeral head articular cartilage by anatomic cross section using computer-aided image analysis software. Sixteen adult cadaveric humeral heads were analyzed. Our findings reveal that the thickness of humeral articular cartilage is substantially thinner than articular cartilage found in the knee. The cartilage is thickest in the central portion of the head and becomes progressively thinner towards the periphery. Surgical techniques used to treat pathology in the glenohumeral joint, specifically thermal energy or mechanical debridement, may have deleterious effects on the relatively thin humeral articular cartilage. PMID- 19292262 TI - Intraosseous glomus tumor in acromion process of scapula. AB - A 25-year-old woman presented with a 5-year history of left shoulder pain, which was constant dull aching in nature, non-radiating with no relieving or aggravating factors and no seasonal variation, gradually increasing in intensity over years. There was no history of trauma or constitutional symptoms. the patient had taken anti-tubercular drugs for 6 months for this pain with no relief. There was point tenderness over the left acromion process. Local temperature was not raised. Movements of the left shoulder were not effected. All routine hematological investigations were normal. Surgical exploration of the lesion revealed a normal soft tissue periosteum and cortex. On removing the cortical bone a red colored jellified tissue was isolated and thoroughly curetted. Histopathology revealed intraosseous glomus tumor. The patient became pain free immediately postoperatively and continued to be symptom free at 24 month follow-up. Glomus tumor located within bone is rare. Only 22 cases of primary intraosseous glomus tumor have been reported in world literature. Glomus tumors are generally treated by meticulously shelling out the entire lesion. Recurrence of symptoms and the need for reoperation have been reported between 12% and 24%. Most authors assume the recurrence is due to inadequate excision, prompting some to recommend more extensive en bloc excision. Tuberculosis being endemic in this region, antituberculer chemotherapy is generally started on clinical suspicion and tissue diagnosis is only attempted in nonresponders. This case however re-emphasizes the value of tissue diagnosis especially when a lesion is at an unusual site like the acromion process. PMID- 19292263 TI - Intracranial metastasis from synovial sarcoma. AB - Synovial sarcomas comprise between 5% to 10% of all soft tissue sarcomas in adults. It has a predilection for young adults and is one of the more frequently misdiagnosed soft tissue sarcomas. It is not unusual for synovial sarcoma to present initially after a traumatic event. It most commonly occurs in close proximity to a large joint. The most common site for metastasis is the lung, followed by lymph node involvement. Synovial sarcoma rarely metastasizes to the skeleton and when it occurs, it most commonly involves the long bones. Cranial metastasis is rare and has only been described in 2 previously reported cases. Reports of other sarcomas having intracranial metastasis include rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, Ewing's sarcoma, and clear cell sarcoma from the kidney. The synchronous or metachronous development of >or=2 primary soft tissue sarcomas has been reported. These are primary soft tissue sarcomas occurring at multiple soft tissue sites without pulmonary or lymphatic involvement. It is often difficult to distinguish between the synchronous or metachronous appearance of a second primary and soft tissue sarcoma metastasis. This article presents a case of a 17-year-old adolescent boy who presented with simultaneous enlarging masses involving the skull and thigh. Open biopsies confirmed synovial sarcoma in both regions. Staging studies, including computed tomography (CT) of his chest, abdomen/pelvis and bone scan were otherwise negative for metastasis. PMID- 19292264 TI - Prophylactic stabilization of proximal femoral metastatic bone disease: reconstruction intramedullary nail fracture. AB - Pathological fracture is an unfortunate manifestation of metastatic bone disease and whenever possible, prophylactic stabilization is preferable to fixation of fracture. The Russell-Taylor reconstruction intramedullary nail (Smith & Nephew Richards, Inc., Memphis, Tennessee) provides a stable construct for the prophylactic management of impending pathological fracture. Implant failure with this system typically involves distal locking screws. Cases of proximal screw cut out and pullout have been reported as has breakage of both proximal locking screws. We present a case of failure of the Russell-Taylor delta reconstruction nail implant, just distal to the point of insertion of the proximal locking screws. PMID- 19292265 TI - Total en bloc spondylectomy in the treatment of aggressive osteoblastoma of the thoracic spine. AB - Osteoblastoma is an uncommon, benign osteoblastic tumor accounting for <1% of all primary bone tumors. Aggressive osteoblastoma is a variant of osteoblastoma and is considered to be a borderline or intermediate osteoblastic tumor, first defined by Dorfman and Weiss in 1984. As a rare tumor, its true incidence and distribution are not clearly known. Treatment of osteoblastoma is complete surgical excision by either curettage or en bloc technique. Recurrence is seen in 10% to 15% of cases, and aggressive osteoblastoma are more likely to recur (75% of the cases. The quadriceps, iliopsoas, and gluteal muscles are most commonly affected. Multiple reports addressed the optimal treatment of tropical pyomyositis. Traditionally, surgical treatment was advocated. However, there are reports of successful nonsurgical or minimally invasive treatment. Based on our experience, it is suggested that the sequence of events in the development of tropical pyomyositis is an initial insult such as vigorous exercise, leading to elevated compartment pressure, resulting in a compartment syndrome. If simultaneous bactermia occurs, the vulnerable muscle is seeded with the infectious organism and abscess formation follows. That may explain other authors' findings that early aspiration, combined with antibiotic treatment, may be sufficient to decompress the muscle, prevent abscess formation and result in full recovery. PMID- 19292271 TI - Priapism as a complication after total hip arthroplasty: a case report and review of the literature. AB - Priapism is defined as a prolonged engorgement or erection of the penis or clitoris that is unrelated to sexual arousal. Recent studies have determined the incidence of priapism to be between 1.5 and 2.9 per 100,000 person years. The incidence of priapism following elective orthopedic surgery is rare. Defining the etiology of a case of postoperative priapism following orthopedic surgery can be difficult, and many times cannot be determined. This case represents the first report of priapism following an elective total hip arthroplasty (THA) performed under spinal anesthesia, and focuses on a review of the literature and potential etiologies of this rare complication. A 44-year-old man with a history of right hip osteoarthritis underwent elective minimally invasive cementless right THA under spinal anesthesia (1 mg of Intrathecal Morphine and 100 mg of intravenous Fentanyl) without initial complication. A preoperative Foley catheter was placed without incident and the patient was placed into the left lateral decubitus position. Surgical time was recorded at 60 minutes. Nine hours after the Foley catheter was removed, the patient developed a persistent painless erection. Successful emergent treatment included a penile Winter shunt, to irrigate blood from the corpora. A computed tomography scan of the pelvis ruled out pelvic deep vein thrombosis as an etiology. This case represents the first report of priapism after an elective THA. The incidence of priapism as a complication is well described and has been associated with numerous medical conditions discussed in this article. PMID- 19292272 TI - Cup-neck impingement due to the malposition of the implant as a possible mechanism for metallosis in metal-on-metal total hip arthroplasty. AB - The metal-on-metal bearing total hip prosthesis is expected to reduce the risk of debris-related osteolysis. However, several reports demonstrated that the socket stem impingement in the metal-on-metal prosthesis due to the implant malposition results in titanium wear debris and secondary metallosis. In this article, we presented a case of massive metallosis due to metal-on-metal impingement. A 60 year woman had severe hip pain due to fracture of the greater trochanter. We planned a revision of the metal-on-metal inlay. Intraoperatively, the trochanteric bursa and joint space were found to be stained black. Black stained granulation tissue was observed between the femoral stem and the great trochanter. Intraoperatively, notching was noticed on both the posteroinferior aspect of the neck of the femoral component and the anterior aspect of the metal liner and polyethylene core. The notch corresponded to the position of impingement between the socket and the femoral neck during the maximum extension of the hip. To clarify the mechanism of cup-neck impingement, the alignment of the prosthesis and pelvic tilt were evaluated. The cup was placed in too much anteverted position. In addition, increased posterior tilt of pelvis in the standing position made the anteversion of the acetabular cup more significant, which enhanced the cup-neck impingement during the gait. Careful attention is necessary for implant alignment and pelvic tilt especially in metal-on-metal bearing total hip arthroplasty. PMID- 19292273 TI - Chondroblastoma of trapezium with metacarpal involvement. AB - Chondroblastomas typically affect the epiphysis of the long bones in young patients, and only rarely occur in the elderly >70 years. Approximately 10% to 20% of chondroblastomas are located in the small bones of the hands or feet, however, they are rare in the carpal bones. Chondroblastoma is a benign tumor, but it may show not only aggressive clinical course but also pulmonary metastases. We report a case of chondroblastoma in a 75-year-old woman that originated in the right trapezium, involving the proximal half of the first metacarpal. She had noticed a mass in her right wrist for approximately 1 year before presenting to the hospital. Radiologically, a large lytic tumor was observed in the right trapezium, with marked expansion of the cortex. On magnetic resonance imaging and computed tomography, the tumor extended from the trapezium into the proximal half of the first metacarpal. Resection of the trapezium and a half of the first metacarpal was performed. Histologically, typical chondroblastoma with small foci of secondary aneurysmal bone cyst was noted. This is an unusual case of chondroblastoma, originating in the carpal bones of an elderly individual, and involving adjacent bone. Benign chondroblastomas of unusual sites may show locally aggressive features, and benign chondroblastomas may occur in elderly persons. PMID- 19292279 TI - Estimating the elbow carrying angle with an electrogoniometer: acquisition of data and reliability of measurements. AB - In clinical practice, the carrying angle of the elbow is assessed by radiographs or goniometers to evaluate the reduction of distal humerus or radial head fractures and epicondylar pathology. This study evaluated a noninvasive technique for estimating the carrying angle in full extension using a 3-dimensional digitizer. Data acquisition was performed on 37 healthy adults. Mean carrying angle was 12.7 degrees +/-3.8 degrees. No side or gender differences were observed (P>.05). Interoperator repeatability was good (interclass correlation coefficient, 0.7), and intraoperator repeatability was excellent (interclass correlation coefficient, 0.85).The major advantage of this technique is the possibility of being adapted to commercial digitizers and being used in new computer technologies for better qualifying anatomic and functional features of the elbow joint. PMID- 19292280 TI - Reconstruction of large defects of the long bones with free vascularized bone grafts: functional results after minimum 5-year follow-up. AB - This study evaluated the clinical and radiographic results of 44 patients (26 men and 18 women) who underwent reconstruction of large defects of the long bones with free vascularized bone grafts. Mean patient age was 29+/-15 years and mean follow-up was 8.6+/-2.1 years. Mean Enneking index was 78.6% (95% confidence interval, 73.8-83.4). No differences were noted for different types of graft, etiologies, or anatomic locations. Regression analysis revealed no association between defect size and functional results. Sixteen patients developed 29 complications. All but 1 patient showed full incorporation of the graft. These results confirm the value of vascularized grafts for bridging large bone defects. PMID- 19292281 TI - Comparison of patient satisfaction with incision cosmesis after standard and minimally invasive total hip arthroplasty. AB - This study evaluated patient assessments and attitudes regarding incision cosmesis following standard and minimally invasive total hip arthroplasty 1 to 3 years postoperatively. A cosmesis questionnaire designed to elicit a score evaluating scar satisfaction was mailed to patients. Although the difference in total cosmesis score between the standard and minimally invasive groups was not statistically significant, patients with a standard incision had better scores at <1 year. One significant finding was worse responses in the minimal incision patients regarding sinking and curling of scar edges. PMID- 19292282 TI - Coralline hydroxyapatite in complex acetabular reconstruction. AB - This retrospective study examined whether a coralline hydroxyapatite bone graft substitute adequately repaired bone defects during complex acetabular reconstructions. Seventeen patients who underwent acetabular revision using Pro Osteon 500 were assessed to determine whether any cups required re-revision, whether bone had incorporated into the coralline hydroxyapatite grafts, and whether the coralline hydroxyapatite grafts resorbed with time. At latest follow up, no cups required re-revision, but 1 had failed. Radiographic evidence of bone incorporation was observed in every coralline hydroxyapatite graft. Graft resorption was not observed. PMID- 19292283 TI - A proximally coated beaded stem minimum 5-year follow-up. AB - This study reports the minimum 5-year clinical and radiographic outcome of a proximally beaded stem. A consecutive series of 159 primary uncemented total hip arthroplasties in 153 patients were evaluated clinically and radiographically by prospectively determined criteria. Radiographic results showed 97% of the femoral stems are bone ingrown and stable by Engh criteria. There have been 5 femoral revisions (2 for subsidence, 2 for loosening, and 1 broken trunion). At minimum 5 year follow-up, this femoral stem has demonstrated reliable clinical function, bone ingrowth, and proximal bone preservation. PMID- 19292284 TI - Open anterior repair without routine capsulorraphy for traumatic anterior shoulder instability in a community setting. AB - Anterior shoulder instability repairs often are performed but rarely reported in community practices. This study assessed outcomes after open anterior repairs without routine capsulorraphy by a community surgeon. Repairs were performed in 64 consecutive shoulders; patient self-assessment questionnaires were mailed to all patients after a minimum follow-up of 2 years. Eighty-three percent reported excellent or good modified Rowe scores at mean follow-up of 5 years. Patients who reported 100% elevation had better outcomes than those who did not report 100% elevation, whereas patients who reported 100% external or internal rotation had comparable outcomes to patients who did not report 100% rotation. Open anterior repair without routine capsulorraphy yielded good outcomes in this community setting. PMID- 19292285 TI - Improvement in documentation of compartment syndrome with a chart insert. AB - To improve documentation of compartment syndrome, an educational program was instituted and a chart insert consisting of a preprinted checklist of history and physical examination parameters for at-risk patients was created. From October 2004 to May 2005, a total of 45 consecutive at-risk patients were identified. Progress notes were divided into group 1 (educational program alone) and group 2 (educational program and checklist). Group 2 showed more complete documentation than group 1. The combination of a chart insert and an educational program proved to be more effective than an educational program alone for improving the documentation of compartment syndrome. PMID- 19292286 TI - Prevalence of methicillin-resistant Staphylococcus aureus among orthopedic patients at a large academic hospital. AB - Community-based methicillin-resistant Staphylococcus aureus (MRSA) contributes to postoperative surgical site infections, and it is therefore important to eliminate nasal carriage of MRSA before surgery. A total of 678 nasal swabs were performed on elective orthopedic patients undergoing surgery with the usage of metal implants. Thirty-eight specimens (5.6%) were positive for MRSA and 146 (21.5%) were positive for methicillin-sensitive S aureus (MSSA). A slow increase in the number of MSSA was noted between 2006 and 2007. Positive cases of MRSA nasal carriage were treated with nasal mupirocin ointment and chlorhexidine baths or showers for 5 days prior to surgery. PMID- 19292287 TI - Short and long term failure of the modular oncology knee prosthesis. AB - This retrospective study investigated the modes of implant failure in 80 patients with modular oncology knee prostheses. Twenty patients (25%) required revision: 12 (60%) for stem loosening, 6 (30%) for bearing failure, and 2 (10%) for infection. Patients with bone sarcomas survived longer; however, long-term prosthetic survivorship was a problem. A higher failure rate was found in patients with tibial tumors and with adjuvant treatment of chemotherapy. This study demonstrates for improved long-term survivorship of modular oncology knee prostheses, there must be improvement in the methods of stem fixation, prosthetic materials, and bearing mechanics. PMID- 19292288 TI - Proximal femoral endoprosthesis for the treatment of metastatic. AB - Traditional reconstructive options may not always be adequate to treat the extensive bone loss that can occur with metastatic disease of the proximal femur. Another method of treatment is resection of the proximal femur and reconstruction with an endoprosthesis. However, the more extensive surgery raises concern for a higher perioperative complication rate in this potentially medically unstable population. This study reviewed 57 patients with metastatic disease treated with 58 proximal femoral endoprostheses. The only perioperative complications were 2 symptomatic deep venous thromboses. Late complications included 3 dislocations, 2 deep venous thromboses, 1 pulmonary embolism, and 4 infections. Three deaths occurred during the perioperative period, all from underlying cancer. Proximal femoral endoprostheses offer a safe treatment option for patients with extensive metastatic disease. PMID- 19292289 TI - A comparison between total hip replacement for osteonecrosis and degenerative joint disease. AB - There has been some reluctance to perform total hip replacement on younger patients with osteonecrosis because of the poor results reported in the earlier literature. To determine the extent to which advances in surgical technique and prosthesis design have led to improved results, we evaluated 203 total hip replacements performed for osteonecrosis and compared them to 300 performed for degenerative joint disease. The failure rate for all femoral and acetabular components combined was 3.7% (22 of 600), in degenerative joint disease and 10.1% (41 of 406) in osteonecrosis. However, results with the most recently used acetabular component, the noncemented HGP-1, were markedly improved over earlier components in both conditions, with only 1 failure in 235 hips (0.4%). The increased survivorship of THR in osteonecrosis with improved component design and surgical technique is encouraging. Total hip replacement is a good option for patients with advanced stages of osteonecrosis, and similar surgical indications should be used for patients with degenerative joint disease and other disorders. PMID- 19292299 TI - Osteochondritis dissecans of the medial femoral condyle associated with malformation of the menisci. AB - Osteochondritis dissecans is a form of osteochondrosis limited to the articular epiphysis and the relationship of osteochondritis dissecans and discoid lateral meniscus has been clearly identified. This article presents a 10-year-old boy with osteochondritis dissecans of the medial femoral condyle associated with hypoplastic medial and partial deficient lateral menisci. The patient presented with activity-related pain and intermittent swellings of his left knee for 2 months without any evidence of significant knee trauma. Magnetic resonance imaging revealed anomalous medial meniscus mimicking displaced bucket-handle tear and partially deficient lateral meniscus with osteochondritis dissecans at the lateral aspect of medial femoral condyle without any significant bone loss with a non-dislocated fragment. Arthroscopy of the knee revealed an incomplete separated osteochondral flap with partial discontinuity that was unstable on probing on the lateral aspect of the medial femoral condyle. The medial meniscus was hypoplastic without a body and only the meniscal rim could be seen. The lateral meniscus had an anomalous vertical insertion of the posterior horn, normal body, but an absent anterior horn. The anterior cruciate ligament was intact, but looked like a peacock's tail. Arthroscopic fixation of the chondral lesion was performed. At 30 months follow-up, the boy had no limitation in his daily and sports activity. The present case is the first description of congenital anomaly of the both menisci and osteochondritis dissecans together. Meniscal or ligamentous anomalies of the knee may be associated with osteochondritis dissecans. PMID- 19292300 TI - Infantile tibia vara: correction of recurrent varus deformity following epiphyseolysis. AB - Infantile tibia vara (infantile Blount's disease) is a condition in which progressive varus of the knee develops due to diminished physeal growth of the medial aspect of the proximal tibia. A 6-year-old boy with severe bilateral infantile tibia vara (Langenskiold stage V) underwent operative correction of his deformities. Surgery consisted of epiphyseolyses of the medial proximal tibiae and valgus-producing corrective tibial osteotomies. Nearly 5 years after symmetric growth, he again developed progressive varus involving his left knee. Usually, a repeat proximal tibial osteotomy is required along with consideration for completion of epiphyseodesis. In this case report, a simpler approach using staples was taken to successfully regain correction in a patient with recurrent deformity who was treated previously by a combination of epiphyseolysis and proximal tibial osteotomy. Stapling of the lateral aspect of the physis allowed the limited remaining growth potential within the abnormal medial physeal region to correct the angular deformity. A small overcorrection into valgus corrected after subsequent removal of the staple. At 14 years, with clinical and radiologic alignment normal, bilateral proximal tibial epiphysiodeses were performed. The patient returned to playing football at a high school level and had no functional limitations. To our knowledge, this is the first report of growth modulation (physeal stapling) resulting in successful correction of recurring genu varum in a knee that had previously undergone epiphyseolysis of the medial proximal tibial physis in advanced infantile tibia vara. PMID- 19292301 TI - Transient osteoporosis of the knee. AB - A 52-year-old man presented with a 3-week history of right knee pain, significant enough to prevent weight bearing. There was no history of knee injury or disease. On knee examination, an effusion and medial joint line tenderness was noted. Knee range of motion was full, but painful. Radiographs revealed minor osteoarthritic changes. Laboratory values were within normal limits, including those associated with infection. Magnetic resonance imaging showed decreased signal intensity on T1-weighted images within the medial femoral condyle and corresponding increased signal intensity on T2-weighted consistent with bone marrow edema as well as edema to the adjacent soft tissues. Bone scanning showed increased uptake of radioisotope in the medial femoral condyle. The patient was diagnosed with bone marrow edema syndrome, and managed medically with analgesics and continued non weight-bearing status. At the 1-month follow-up, his symptoms had improved significantly. The patient progressively increased weight bearing as tolerated over the subsequent weeks. At 12-week follow-up, symptoms had completely resolved. Magnetic resonance imaging showed no signal abnormalities in the affected knee. Transient osteoporosis is an uncommon and typically self-limiting condition that can be challenging to diagnose and treat. The clinician must be aware of this entity, as well as its typical course, to prevent unnecessary testing and invasive intervention. PMID- 19292302 TI - Tibial tubercle fragmentation: a clue to simultaneous patellar ligament avulsion in pediatric tibial tubercle fractures. AB - Simultaneous avulsions of the tibial tubercle and patellar ligament have been reported, but are rare. We present an 11-year-old boy who was initially diagnosed with a Type IIIA tibial tubercle avulsion fracture after falling off of his bicycle. Intraoperatively, following exposure and fixation of the tibial tubercle fragment, knee range of motion under live fluoroscopy revealed a stationary patella with no tension in the patellar ligament. The incision was extended and a distal avulsion of the patellar ligament from the tibial tubercle was identified, with a small flap of tibial tubercle periosteum remaining attached. Transosseous suture fixation of the avulsed periosteal flap was achieved with 2 No. 5 ethibond sutures placed through connecting drill holes. Postoperatively, the patient was placed in a long leg cast with the knee in extension for 6 weeks. The injury to healed and the patient returned to full activities, although he required 2 courses of physical therapy and 4.5 months to regain full range of motion. Preoperative diagnosis of simultaneous tibial tubercle fracture and patellar ligament avulsion can be difficult. Palpation of the patellar ligament for gaps may not be possible due to a large knee effusion. Most patients do not tolerate quadriceps testing on examination and do not have patella alta on radiographs. In retrospect, a clue to this diagnosis in our patient was the preoperative radiographic finding of multiple calcified fragments below the patella. The calcified fragments likely represent the avulsed tibial tubercle periosteum attached to the distal patellar ligament. In addition, the large tibial tubercle fragment is separated and rotated superiorly, a finding that was also noted in two other similar case reports. PMID- 19292303 TI - Delayed posttraumatic pseudoaneurysm of the anterior tibial artery mimicking a malignant tumor. AB - The anterior tibial artery arises at the lower border of the popliteus muscle, and passes anteriorly to lie on the interosseous membrane medial to the fibular neck. Injury to the artery results in extravasation of blood, and the tamponade effect of the surrounding tissue may contain acute hemorrhage. The blood clot gradually liquefies resulting in the formation of a fibrous capsule. Consequently, a pseudoaneurysm contains no normal elements of arterial wall, as opposed to a true arterial aneurysm. Pseudoaneurysm of the anterior tibial artery has been reported as a complication of fracture surgery and is usually felt to be iatrogenic. It has occurred following a fracture treated without surgery and may be related to the vessel being tethered by the fascia and interosseous membrane. It has also been reported in cases of blunt trauma to the leg without fracture, and as well following elective orthopedic procedures. We report the case of a patient 7 months following open reduction and plate fixation of a fracture of the distal third of the tibial shaft, who presented with a large painful proximal leg mass thought to be a malignant tumor. Three previous such presentations have been found in the literature and illustrates the importance of suspecting a delayed vascular injury following lower leg trauma. Duplex ultrasound examination should be performed initially to avoid an inadvertent incision and catastrophic bleeding as might occur with an inappropriate biopsy. Preoperative and intraoperative angiograms are essential to confirm the diagnosis, delineate the site and type of injury, and help to plan the treatment. PMID- 19292304 TI - Tuberculous tenosynovitis of flexor digitorum longus tendon. AB - Extrapulmonary tuberculous involvement of musculoskeletal system accounts for approximately 1% to 5% of cases. Tuberculous involvement of the feet tendon is even rarer. We are reporting an extremely rare case of tuberculous tenosynovitis of flexor digitorum longus tendon in a 68-year-old man. A 68-year-old farmer from India presented with a painless swelling over medial aspect of left ankle for 3 months. Examination of left ankle and foot revealed non-tender, fluctuant swelling, anteroinferior to medial malleolus. Plain radiographs of the ankle and foot revealed no abnormality. Laboratory findings, including erythrocyte sedimentation rate, were within normal limits. When excision was attempted, purulent material came out on incising the subcutaneous tissue. Gram staining, acid fast bacilli staining and pyogenic culture detected no organism. Histopathological examination of the granulation tissue showed non-specific inflammation. The wound healed well with secondary intention over a 15-day period. Two months postoperatively, swelling recurred at the same site. Magnetic resonance imaging showed small hypointense collection in the tendon sheath of left flexor digitorum longus behind the left ankle joint and adjacent lower end of tibia on the T2-weighted, STIR, and gradient (T2 with fat saturation) images, which appeared hypointense on T1-weighted images, suggestive of tenosynovitis. Tenosynovectomy was performed. Mycobacterium tuberculosis was isolated on culture. Histopathological examination, this time revealed epitheloid granuloma. Patient recovered uneventfully after 5-drug antituberculous regime. PMID- 19292305 TI - Melorheostosis: case report with 20-year follow-up. AB - The authors report on a typical case of melorheostosis observed in an 18-year-old man, localized in the II digit of the right hand causing tumefaction and irregular pain. At 20-year follow-up, the patient showed a radiographical slight worsening of the lesion. Radiographic examination showed a mild increase in cortical and endosteal hyperostosis with bone shape alteration in the II digit of the right hand, slight cortical and endosteal hyperostosis at the level of right shoulder girdle involving the humerus. The other part of the skeleton was not affected. Clinical examination revealed swelling and tumefaction of the forefinger of the right hand and reduced articular motion both in proximal and distal interphalangeal joint and metacarpal phalangeal joint. The reminder of the clinical examination was normal. Melorheostosis is an uncommon mesenchymal disorder characterized by a chondral hyperostosis of the long bones associated to a sclerosis of the spongious bone. The etiology of this affection is unknown: the most accredited hypothesis is a developmental anomaly with a sequential sclerotomes distribution. Both genders may be affected, with the long tubular bones of the lower limbs more involved. This anomaly is often asymptomatic but maybe accompanied by pain with a smooth symptom progression and periodic exacerbations. Therapy is mainly symptomatic and comprises anti-inflammatory drugs or surgical approach for removal of soft tissue that may preclude daily activities. This case report demonstrates that melorheostosis is a relatively benign disorder with mildly progressive course. PMID- 19292306 TI - Combined glenoid and humeral head allograft reconstruction for recurrent anterior glenohumeral instability. AB - Recurrent glenohumeral instability can be a challenging therapeutic problem to orthopedic surgeons. Oftentimes, recurrent anterior glenohumeral instability is associated with a host of capsular, labral, and occasionally, osseous deficiencies. Current treatment strategies for significant bony deficiency within the shoulder are aimed at nonanatomic restoration of glenohumeral anatomy with resultant limitation in range of motion parameters. While these techniques may prevent recurrence of instability, the resulting loss of motion and decreased function may not be suitable for young, active individuals. This case report presents a patient with recurrent anterior glenohumeral instability from a combined, large glenoid and humeral head bone deficiency. A novel surgical technique of restoring both humeral and glenoid bone is introduced as a way to prevent decreased motion after surgery while still maintaining stability. This reconstruction offers a more anatomic restoration of bone deficiency with the desired effect of achieving a more complete postoperative range of motion, while still conferring stability. The patient has returned to activities of daily living. PMID- 19292307 TI - Osteoid osteoma of the hamate. AB - Uncommonly affecting the carpus, osteoid osteoma, a benign bone lesion, will involve the hand in 10% of cases. With negative initial radiographs and persistent tenderness in a 15-year-old boy after a fall, a wrist magnetic resonance imaging revealed extensive hamate edema without a fracture line. He was casted for a presumed hamate fracture. Without complaints of dysthesias or weakness, marked localized tenderness over the hamate, mild dorsal carpal edema, and limitation of wrist motion was noted 6 months later despite immobilization and rest. A computed tomography (CT) scan revealed a round, osteolytic lesion ulnarly at the hamate hook's base. Within the classic zone of sclerosis, a central calcification was visualized. Irregular periosteal bone formation is seen on the hamate hook and the dorsal aspect of the hamate. A volar curetting of the lesion was performed without grafting as stability was not threatened. Permanent sections consisted of a bony nidus with compact sclerotic, poorly organized osteoid. The central tissue was vascular with many osteoblasts, fibroblasts and mesenchymal cells. Following surgery, his pain remitted completely. Characteristics of pain and swelling have been consistent among reported cases of osteoid osteoma. Presentation may include warmth, erythema and restriction of motion. A delay in diagnosis is present in all cases involving the hamate usually due to negative plain radiographic findings. Magnetic resonance imaging findings further delayed diagnosis and treatment. Lesions are more accurately and frequently diagnosed in children with the use of CT. PMID- 19292308 TI - A case of factitious subfascial emphysema. AB - A 44-year-old woman with a history of borderline personality disorder, Hepatitis C, and multiple hospital admissions for cellulitis and cutaneous abscesses presented with pain of several days duration in her left upper extremity following getting her left arm "stuck" in a subway turnstile. The pain and swelling had progressively worsened following the injury. At the time of presentation the patient's temperature was 98.6 degrees F, heart rate was 82 beats/minute, blood pressure of 116/60, and an oxygen saturation of 98% on room air. The left upper extremity was diffusely swollen but minimally tender throughout. Crepitus was noted along the left arm, elbow and forearm areas. No open wounds were visualized during the examination. Radiographs and computed tomography revealed diffuse subcutaneous emphysema extending from the left axillary region to the wrist, as well as gas beneath the fascia extending between multiple muscles in both the upper and lower areas of her left upper extremity. The possibility of necrotizing fasciitis or gas gangrene prompted immediate operative exploration. No evidence of infection was found during the exploration. Stat gram stains sent during the procedure revealed no organisms. Aerobic, anaerobic, fungal, and acid-fast bacteria cultures taken during the operation revealed no final growth. The patient was stable throughout the hospitalization, remaining afebrile with a normal white blood cell count. The gas in the subcutaneous and subfascial layers was not caused by infection but presumably was due to factitious self-injection of air. PMID- 19292309 TI - Rare case of upper arm compartment syndrome following biceps tendon rupture. AB - We present a case of upper arm compartment syndrome following a biceps tendon rupture in a 77-year old man on warfarin sodium. Compartment syndrome is common in the forearm and leg, but rare in the upper arm with only a handful of cases reported in the literature. Our patient's anticoagulated state predisposed him to the development of compartment syndrome. To the best of our knowledge there has been only one other case reported in the literature of upper arm compartment syndrome following biceps tendon rupture in a patient on warfarin sodium. Compartment syndrome of the upper arm is a rare occurrence. Previous cases have occurred due to malposition of blood pressure cuffs, injections, venepuncture, trauma, tourniquets, shoulder dislocation, surgical complication, subatmospheric pressure induced, biceps rupture, and triceps rupture. The fascia of the upper arm is relatively thinner and more distensible than the fascia of the leg or forearm. This creates more room for the compartment to swell before pressures builds up to a significant level. Thus, a significantly increased amount of pressure needs to build up before compartment syndrome will occur in the upper arm. Once the diagnosis was formed, the patient was treated with emergent fasciotomy and evacuation of hematoma. After a prolonged hospital stay, the patient was released with minor neurological deficits. At final follow-up, the patient was neurovascularly intact with no complaints of numbness or tingling, and he had regained full motor function throughout. PMID- 19292310 TI - External fixation of pubic symphysis diastasis from postpartum trauma. AB - Pubic symphysis diastasis is recognized as a possible complication of pregnancy. When this occurs, pubic symphysis diastasis may cause anterior widening and loss of stiffness within the pubic symphysis, causing potential instability in the pubic joint. The persistent loss of reduction can cause substantial disability in postpartum women. Pubic symphysis diastasis has previously been treated conservatively using a pelvic girdle and bedrest with some success. When a diastasis >3 cm is present, however, surgical intervention may be needed to preserve the integrity of the pubic symphysis joint. To date, most surgical procedures for reduction of pubic symphysis diastasis have been via internal fixation with plates and screws on the superior pubic rami. Although internal fixation provides good structural support, this method would be inadequate if a postpartum pubic symphysis diastasis patient has significant reproductive organ damage, due to the risk of soft tissue infection or osteomyelitis. External fixation is an alternate method of pubic symphysis diastasis treatment that has not received significant attention in the literature to date. We present the following case report to highlight a novel use of a pelvic frame external fixator for treatment of a severe postpartum pubic symphysis diastasis with organ damage. This article outlines a treatment alternative to internal fixation for cases of pubic symphysis diastasis with a contaminated pelvic environment. PMID- 19292311 TI - Decreased range of motion following arthroscopic remplissage. AB - The Hill-Sachs lesion is an impression fracture of the posterolateral margin of the humeral head that commonly results from impaction with the anterior glenoid rim during subluxation or dislocation of the shoulder. This bony defect has been implicated as an etiology of recurrent instability of the shoulder. The "remplissage" technique described by Wolf is an arthroscopic method of filling the Hill-Sachs defect via infraspinatus tenodesis and posterior capsulodesis. The purpose of this technique is to prevent recurrent instability by making the Hill Sachs lesion extra-articular thereby eliminating engagement of the defect with the anterior glenoid rim. In addition, the infraspinatus tenodesis acts as a checkrein by preventing anterior translation of the humeral head. We report a complication of this recently described technique. A 28-year-old man who failed conservative management underwent an arthroscopic Bankart repair with a concomitant remplissage procedure. Two years postoperatively, the patient continued to struggle with the loss of external rotation. Conservative management with physical therapy and range of motion exercises failed. The patient was treated with an arthroscopic release of the tenodesed infraspinatus, which improved external rotation by 202 in circle intraoperatively. At 6 months postoperatively, the patient has maintained a significant increase external rotation. We hypothesize that the loss in external rotation occurred as a result of the tenodesis of the infraspinatus and posterior capsule into the Hill-Sachs defect. The tenodesed cuff and capsular tissue acted as a mechanical block to external rotation of the shoulder. PMID- 19292312 TI - Acute and chronic management of posterolateral corner injuries of the knee. AB - Injuries of the posterolateral corner of the knee are uncommon, but can lead to chronic disability from persistent instability and resultant articular cartilage degeneration if not appropriately treated. These injuries rarely occur alone and are often associated with cruciate ligament tears or tibial plateau fractures. Clinical management is dictated by the severity and timing of injury. Early diagnosis may allow primary repair of the posterolateral corner rather than delayed reconstructon. This review article provides a brief overview of posterolateral corner injuries of the knee, with a focus on clinical management based on grade of injury and timing of diagnosis. PMID- 19292319 TI - Grammont inverted prosthesis for the treatment of cuff tear arthropathy: a 6-year follow-up study. AB - This study evaluated mid-term functional results of the Grammont inverted prosthesis for the treatment of shoulder osteoarthritis associated with massive cuff rupture. Twenty-eight patients with a minimum of 6 years of follow-up were evaluated. Clinical evaluation was performed on all patients preoperatively and at follow-up using the Constant score. Radiologic assessment included radiographs and magnetic resonance imaging preoperatively and radiographs at follow-up. Constant score improved from 27.4 preoperatively to 64.6 postoperatively. The inverted prosthesis is attractive and remains one of the options for treating cuff tear arthropathy. However, glenoid loosening remains a serious problem. PMID- 19292320 TI - Complications of cervical halo-vest orthoses in elderly patients. AB - Halo-vest orthoses have been associated with complications. Previous reports have suggested increased complications in elderly patients; however, data are limited. This study reviewed 75 patients =/>65 years with cervical spine fractures treated with halo-vest orthoses. Forty-one patients (55%) experienced at least 1 complication. Pin-site problems were the most frequent adverse outcome. Seventeen patients (23%) had significant pulmonary compromise. Pulmonary complications were associated with high morbidity. We found an 8% early mortality rate. Halo-vest orthoses are useful devises. However, adverse events experienced by elderly patients are common and may lead to significant morbidity and mortality. PMID- 19292321 TI - Risk factors in total joint arthroplasty: comparison of infection rates in patients with different socioeconomic backgrounds. AB - Infection after total joint arthroplasty is a serious complication. Several risk factors have been shown to increase the risk of total joint infections. The purpose of this study was to evaluate whether socioeconomic background was a risk factor for infection in primary total joint arthroplasty. A retrospective chart review was conducted over a 4-year period on a single surgeon's split practice between private patients with mostly private insurance and Medicare and county based patients with predominately indigent county health coverage and Medicaid. An infection rate was calculated for each population in both primary total knee and hip arthroplasty. The two populations were statistically analyzed for differences in age, preoperative diagnoses, and socioeconomic background. To our knowledge, this is the first study showing an increased risk of infection in total joint arthroplasty based on socioeconomic background. PMID- 19292322 TI - Decreased proliferation of mesenchymal stem cells in corticosteroid-induced osteonecrosis of femoral head. AB - The ability of self-repair in patients with corticosteroid-induced osteonecrosis of the femoral head is limited, and it has been suggested the cause is likely relevant to the poor proliferation activity of mesenchymal stem cells in the femoral head region. This study measured the number and proliferation activity of human mesenchymal stem cells in patients both with and without corticosteroid induced osteonecrosis of the femoral head. Bone marrow was collected from the proximal femur in patients with steroid-induced osteonecrosis of the femoral head (osteonecrosis group, n=18) and patients with new femoral neck fractures without osteonecrosis (control group, n=11). Mesenchymal stem cells were isolated by density gradient centrifugation, and then selected by the adhesive method. The MTT reduction assay method was used to evaluate the level of proliferation. Cells from osteonecrosis patients showed reduced proliferation ability compared with the control patients. The percentage of cells in the S+G2/M phase was decreased significantly (P<.01) in the osteonecrosis group. The decreased proliferation ability of mesenchymal stem cells may play a role in the low repair capacity of steroid-induced osteonecrosis of femoral head. The altered function of mesenchymal stem cells may be responsible for the pathogenesis and progression of osteonecrosis. PMID- 19292323 TI - Impaction grafting for femoral component revision in a goat model using washed morselized cancellous allograft. AB - To determine whether washing morselized cancellous bone allograft in impaction grafting for revision hip arthroplasty would improve mechanical and biologic performance, left hip hemiarthroplasty with a collarless stem cemented into impacted morselized cancellous bone was performed in 22 goats. Washed allograft was used in the experimental group, and standard allograft was used in the control group. One of 11 experimental and 4 of 11 control implants were observed to be loose at 8 weeks. Washing allowed significantly more morselized cancellous bone to be placed in the experimental group compared to the control group (7.7+/ 1.9 and 6.2+/-2.0 g, respectively, P<.05). Significantly less in vivo subsidence over the 8-week study period also was demonstrated in the experimental group compared to the control group (0.4+/-0.4 and 2.2+/-2.3 mm, respectively, P<.05). Angular motion during cyclic +/-1.5 Nm loading demonstrated significant differences between the 2 groups at time zero (2.67 degrees +/-1.02 degrees for the control group and 1.98 degrees +/-0.47 degrees for the experimental group, P<.05) and at 8 weeks (2.40 degrees +/-0.38 degrees for the control group and 1.74 degrees +/-0.55 degrees for the experimental group, P<.05). Histology showed little difference between the 2 groups, but there was a trend toward less inflammation in the experimental group. PMID- 19292324 TI - The tibial sulcus as a reference point for total knee arthroplasty. AB - Determining the amount of tibial resection is an important aspect of knee arthroplasty. The tibial sulcus is a reference point that should be unaffected by degenerative processes. This study compared the tibial sulcus and plateau height for 50 arthritic knees to 50 normal knees on magnetic resonance imaging. The tibial sulcus measured 17.09 mm in arthritic knees and 17.88 mm in normal knees. Sulcus height differences were not statistically significant. Based on these findings, a tibial cut 12 to 14 mm from the sulcus is recommended as a reference point in arthritic knees. This measuring technique has application in limited incision knee arthroplasty. PMID- 19292325 TI - Suture anchor versus suture through tunnel fixation for quadriceps tendon rupture: a biomechanical study. AB - This biomechanical study compared suture anchors versus transosseous sutures for repair of quadriceps tendon ruptures using a force of 150 N at a frequency of 0.5 Hz. No significant difference in displacement was found between the 2 techniques with initial loading or with load or no load after 1000 cycles. Displacement after 1000 cycles for suture anchors and bone tunnels was 4.65 and 4.50 mm, respectively. These findings suggest a possible role for suture anchors in repairing quadriceps tendon ruptures. Suture anchors are relatively expensive but require less dissection over the patella and do not involve suture placement about the patella tendon. PMID- 19292326 TI - Association of knee bone bruise frequency with time postinjury and type of soft tissue injury. AB - This study correlated the frequency of bone bruises with soft tissue injuries in the knee and examined bruise frequency as a function of time postinjury. Magnetic resonance imaging of 1546 patients revealed bone bruises in 19% of patients without an associated meniscal or ligamentous injury. For those patients presenting with at least one meniscoligamentous injury, the frequency of bruising was 60% at 0 to 4 weeks, 42% at 4 to 10 weeks, and 31% at 10 to 26 weeks postinjury. The frequency of bruising varied with the presence of concomitant injuries, with the greatest frequency of bruises (78%) observed in patients with anterior cruciate ligament injuries. PMID- 19292327 TI - Unusual metastases from extremity soft tissue sarcomas. AB - The most common metastatic route for extremity soft-tissue sarcomas is via the venous system to the lungs. Metastases to other sites such as the brain, liver, and soft tissue distant from the primary tumor are rare. A tumor registry, prospectively kept since 1986, was reviewed for unusual metastatic spread. Of 3671 tumors, 346 high-grade extremity soft-tissue sarcomas were evaluated. A total of 15 patients (4.3%) presented with initial recurrence of disease that was extrapulmonary and distant from the site of the primary tumor. Four of these patients (27%) were successfully treated for their recurrence. Based on these findings, a different strategy for follow-up of patients after treatment of a high-grade extremity soft-tissue sarcoma is suggested. PMID- 19292328 TI - The use of locking plates in orthopedic oncology reconstructions. AB - Locking-plate systems are believed to provide better purchase in poor quality bone and equivalent purchase with fewer screws, and also to limit screw pullout by functioning as fixed-angle devices. This retrospective study examined 25 oncologic reconstructions involving locking plates. There were 8 cases of open reduction and internal fixation for pathologic fracture or nonunion and 17 limb salvage reconstructions. Mean follow-up was 18.2 months with 92% of constructs intact (there were 2 implant-related failures). Locking plates offer advantages that can be useful in orthopedic oncology reconstructions. The long-term performance and mechanisms of failure of these implants remains to be defined. PMID- 19292331 TI - Subtrochanteric fracture in a patient with trabecular metal osteonecrosis intervention implant. AB - We present an unreported case of a sixty-four year old woman who suffered a subtrochanteric fracture of the femur four weeks after implantation of a trabecular metal osteonecrosis intervention implant for osteonecrosis of the femoral head. Traditional treatments for femoral head avascular necrosis include observation, core decompression, bone grafting, vascularized fibular grafting, osteotomy, hemiarthroplasty and total hip arthroplasty. With the recent development of trabecular metal, a new implant system has been developed using the properties of trabecular metal for treatment of osteonecrosis. Trabecular metal is a relatively new and unique material that physically and mechanically more closely resembles bone than any other prosthesis. Its use in osteonecrosis of the femoral head has shown promising results with few if any short term complications. To the best of our knowledge, besides normal progression of the disease, no major complications of this implant system have been reported. This article presents the first reported case of a subtrochanteric fracture of the femur following implantation of a trabecular metal osteonecrosis implant. PMID- 19292332 TI - Traumatic simultaneous asymmetric hip dislocations and motor vehicle accidents. AB - Hip dislocations are becoming more frequent with increasing numbers of motor vehicle accidents. Bilateral hip dislocations are a rare injury, and simultaneous asymmetric dislocations are even more rare. Of the 20 cases described in the English literature, 16 have been sustained by occupants of motor vehicle accidents. This article presents the first case of a female with traumatic simultaneous asymmetric hip dislocations, as well as the first patient with associated asymmetric acetabular wall fractures. PMID- 19292333 TI - Traumatic lower extremity arteriovenous fistulae in children. AB - Traumatic arteriovenous fistulae are rare injuries in the pediatric population. Most are caused by penetrating injuries or are post-surgical in nature. Fistulae resulting from non-penetrating injuries are often missed early in the course of physical examination. This occurs due to the absence of clinical signs of arterial or venous injury, despite the close proximity of the affected vessels to point of injury. Likewise, signs and symptoms of post-surgical vascular injury may be difficult to discern from normal postoperative discomfort. The astute clinician must be on alert for unusual presentations of vascular injury to intervene in an expeditious manner. This article presents a series of vascular complications following either blunt injury or surgical management of the lower extremity in children who presented to our facility between November 2004 and December 2005. PMID- 19292334 TI - Conservative follow-up after epidural abscess and diskitis complicating instrumented metal interbody cage. AB - There have been few reports on the imaging diagnosis and treatment of infection of interbody cages used in posterior lumbar interbody fusion. Even in a patient undergoing posterior lumbar interbody fusion using a metal cage, magnetic resonance imaging (MRI) might be able to diagnose epidural abscesses and diskitis when titanium alloy implants are used. When infection of the instrumented cage is diagnosed, emergency debridement with or without removal of implants, including the cage, appropriate wound closure, and the administration of potent antibiotics is the standard treatment. This article describes the conservative follow-up of a patient with epidural abscess and diskitis following instrumented posterior lumbar interbody fusion using a metal cage without surgical intervention. A 40 year-old man had undergone instrumented posterior lumbar interbody fusion at the L5/S1 level approximately 1 year previously. He suddenly developed a fever (temperature, 38 degrees C) and severe lower back pain without apparent cause. Late infection was suspected from his symptoms and laboratory findings, and a wide spectrum antibiotic was administered. On MRI, the low and high signal areas near the end-plate of L5/S1 were detected by T1- and T2-weighted imaging, respectively, and an abscess was noted around the cage. We recommended debridement as treatment; however, he refused further surgical intervention by reason of symptomatic remission. Therefore, wide spectrum antibiotics were continued. Fortunately, no lower back pain has recurred. Examination by MRI during the clinical course clarified the presence and disappearance of an epidural abscess and diskitis. Although the outcome of this case was fortunate, this conservative treatment is unusual, and is not recommended as standard treatment. PMID- 19292335 TI - Chondromyxoid fibroma of the lumbar spine in a pediatric patient. AB - Chondromyxoid fibroma is a rare nonmalignancy that comprises <1% of all bone tumors. It typically presents with pain, swelling, and tenderness to palpation. The lesion has a predilection for the metaphysis of long bones of the lower extremity, most commonly in the proximal tibial metaphysis. Patients often present in their second or third decade of life, although some reports have included a younger average age. Sporadic reports of chondromyxoid fibroma in the spine are found in the literature. This article presents the second case of a chondromyxoid fibroma of the lumbar spine in a pediatric patient, along with a literature review with emphasis on recurrence rates and malignant transformation. PMID- 19292336 TI - Management of elbow ankylosis in a head injured patient with floating elbow injury. AB - The development of heterotopic ossification around the elbow has been linked to trauma (particularly fractures or dislocations), closed head injury, spinal cord injury, burns of the extremity, genetic traits, adult respiratory distress syndrome, and orthotopic liver transplant. Pain and lack of motion are the most common complaints during the progression of ectopic bone about the joint. Opposite to common credence, fracture healing is not accelerated in patients with traumatic brain injury. Formation of heterotopic bone is not a clinically significant form of healing, therefore healing rates of patients with traumatic brain injury should coincide with those of the general population. Heterotopic bone formation on the medial side of the elbow can cause cubital tunnel compression and ultimately result in ulnar neuropathy. The literature is devoid of any report concerning ectopic bone formation in traumatic brain injury with simultaneous bony trauma to the affected arm. This article presents a case of a patient with a chronic rigidly ankylosed elbow secondary to heterotopic ossification, after suffering a traumatic brain injury, complex periarticular elbow fractures, and an ipsilateral segmental forearm fracture. PMID- 19292337 TI - Modified ulnar translocation technique for the reconstruction of giant cell tumor of the distal radius. AB - Giant cell tumors of the distal radius have been frequently described as difficult to treat, chiefly because of their close proximity to multiple tendons, median nerve, radial artery and carpus.The aim of treatment is to remove the tumor completely and preserve the radiocarpal and radioulnar joints.However, this is not always feasible as giant cell tumors seem to behave more aggressively and have a higher recurrence rate in the distal radius, even if local adjuvant treatment with phenolmethylmethacrylate or liquid nitrogen is applied. The above incidence is increased in Campanacci grade III lesions, which are characterized by fuzzy borders, loss of cortical continuity, and extension into soft tissues. In these cases, wide excision instead of intralesional excision may be advocated, particularly when the tumor breaks through the cortex, violates the articular surface, and destroys >50% of the surrounding metaphysis. Several reconstructive options (e.g., resection arthroplasty, prosthetic replacement, arthrodesis, ulnar translocation, centralization of the carpus over the remaining ulna, use of a nonvascularized, or vascularized fibular graft [with or without arthrodesis], and allograft replacement) have been described for the treatment of either recurrent or primary grade III giant cell tumor with destruction of the bone cortex and associated soft tissue mass. Ulnar translocation has been mentioned rarely in the literature, and, according to our knowledge, only 10 cases have been previously reported. This article presents a case of a woman with a grade III giant cell tumor of the distal radius. Wide excision of the tumor followed by reconstruction of the distal forearm with a modified ulnar translocation technique and wrist arthrodesis led to optimum results and no mass recurrence at 13 years postoperatively. PMID- 19292338 TI - Coccidioides immitis osteomyelitis of the radius presenting as Ewing's sarcoma. AB - Coccidioides immitis osteomyelitis is a relatively rare manifestation of a disease that most commonly presents with pulmonary infection. Disseminated disease occurs in approximately 1% of infected individuals, with bony involvement in 10% to 50% of those patients with extrapulmonary infection. Diagnosis and treatment of patients with primarily osteoarticular complaints is frequently delayed, which may result in progression of disease and suboptimal results. This article discusses the successful treatment of a patient whose initial presentation was suggestive of Ewing's sarcoma of the proximal radius, but was found on biopsy to have coccidioidomycosis osteomyelitis. The patient was treated with surgical debridement and systemic as well as intralesional antifungal therapy. PMID- 19292339 TI - Ochrobactrum anthropi septic arthritis of the acromioclavicular joint in an immunocompetent 17 year old. AB - Ochrobactrum anthropi is a ubiquitous, aerobic, gram-negative bacillus of low virulence, most frequently associated with nosocomial infections and infections related to indwelling catheters in immunocompromised hosts. This article presents the first reported case of O anthropi septic arthritis, here occurring at the acromioclavicular joint of an otherwise healthy patient, and provides treatment recommendations based on our experience. PMID- 19292340 TI - Isolated acute dislocation of the proximal tibiofibular joint. AB - Lesions of knee's lateral side are less common than medial injuries. The anatomy of the lateral ligaments and the presence of additional structures (eg, fibula head) can cause diagnostic problems. Isolated dislocation of the proximal tibiofibular joint is unusual; therefore, it may be overlooked in the emergency department. Many cases are missed due to failure of diagnosis. This type of injury is common in athletes whose sports require twisting motions of the flexed knee (eg, wrestling, parachute jumping, judo, gymnastics, skiing, rugby, football, soccer, track, baseball, basketball, racquetball, and roller-skating). Anterolateral dislocation of the proximal tibiofibular joint is seemingly rare in soccer players, as less than a handful cases have been reported in the literature. The diagnosis can be made by clinical examination, plain knee radiographs, and, sometimes, computed tomography (CT) scanning for further confirmation. Treatment usually consists of closed or open reduction. In complicated cases, however, arthrodesis or resection of the fibular head may be required. This article reports a rare case of acute isolated anterolateral dislocation of the proximal tibiofibular joint in a soccer player and discusses the joint anatomy, types of dislocations, mechanisms of injury, and treatment options. PMID- 19292341 TI - Complete and incomplete femoral stress fractures in the adolescent athlete. AB - Stress fractures of the lower extremities are a known risk of intense physical activity. Femoral stress fractures represent a small subset of lower extremity stress fractures but can have grave consequences if undetected, resulting in a displaced fracture. If detected prior to displacement, conservative treatment of these fractures is highly successful. However, displacement frequently makes surgical treatment necessary. Unfortunately these fractures are often difficult to detect. Symptoms of stress fractures are frequently vague, particularly in femoral stress fractures. Specific physical examination techniques can assist in diagnosis but require a very high index of suspicion. Plain radiographs are also frequently negative in early stress fractures, further confounding diagnosis, and bone scan or magnetic resonance imaging studies are frequently needed. We present a case of an otherwise healthy 15-year-old male athlete with bilateral femoral stress fractures. The patient reported bilateral knee pain of 2 months' duration with physical activity, but had not curtailed his activities. His pain had been evaluated with plain radiographs, which were interpreted as normal. Immediately prior to presentation, while playing lacrosse, he sustained a displaced right supracondylar femur fracture. On further evaluation with bone scan he was found to have a contralateral nondisplaced supracondylar femoral stress fracture. His displaced fracture was managed with a retrograde intramedullary nail and his nondisplaced fracture was treated conservatively. PMID- 19292351 TI - Effects of the lateral approach on blood flow of the gluteus medius and abductor function in total hip arthroplasty. AB - Total hip arthroplasty was performed in 19 patients using the lateral approach. Laser Doppler flowmetry was used to measure the blood flow in the anterior and posterior parts of the gluteus medius. The anterior part was split and detached. The posterior part was left intact. The blood flow in the gluteus medius of the anterior part was decreased by 15.4%, while that of the posterior part remained constant during surgery. Postoperatively, the Trendelenburg sign remained positive in 3 of 16 patients. No correlations were observed between a positive Trendelenburg sign and reduction in blood flow of the gluteus medius. PMID- 19292352 TI - Correlation of angulation on initial and healed radiographic views of surgical humerus neck fractures. AB - This retrospective study evaluated the correlation of the angulation on initial and healed radiographs of displaced surgical humerus neck fractures treated nonoperatively, with emphasis on the initial axillary view. Thirty-six patients with displaced isolated surgical humerus neck fractures were treated nonoperatively to union. Fracture angulation of the injury and healed views were measured. Angulation on the initial axillary view was a poor predictor of angulation at union. Initial axillary radiographs of surgical neck fractures treated nonoperatively overestimated final angulation, with the Y view being more predictive of angulation at union. PMID- 19292353 TI - Complications following quadriceps-sparing total knee arthroplasty. AB - This study examined whether quadriceps-sparing total knee arthroplasty (TKA) through a minimal medial incision could be performed without an increased risk of complications. Data were collected prospectively on 209 patients who underwent quadriceps-sparing TKA though a median parapatellar approach. Surgical complications included 2 patellar tendon avulsions, 2 lateral collateral ligament ruptures, and 1 medal collateral ligament rupture. Arthrofibrosis occurred in 21 patients (10%) requiring manipulation under anesthesia, and minor wound complications occurred in 22 patients (11%). Unintended cement retention was noted on 16 radiographs (8%). At 6 months of follow-up, Knee Society scores and functional outcomes were unchanged from previous experience with standard TKA. Compared to previous experience with traditional medial parapatellar approaches, the rate of intraoperative complications such as ligament rupture and retention of cement were higher than expected. Complication rates in the second 100 patients who underwent quadriceps-sparing TKA showed no significant difference compared to previous patients who underwent standard TKA. Complications were more common in patients with preoperative peripheral vascular disease, body mass index >33, and valgus deformity >10 degrees ; these patients may not be good candidates for this modified procedure. This quadriceps-sparing technique required a progressive learning curve and has not yet proven to be superior to standard approaches. However, complication rates after extensive experience were not significantly increased. PMID- 19292354 TI - Sagittal alignment as a risk factor for adjacent level degeneration: a case control study. AB - This study examined whether sagittal alignment, preexisting adjacent level degeneration, and smoking predispose patients to adjacent segment degeneration following lumbar fusion. Fifty-one patients with adjacent segment degeneration were identified and matched with control patients based on age, sex, level, and date of index surgery. Preexisting adjacent level degeneration and sagittal alignment through the fusion and from L1-S1 were determined before and after initial surgery. Patients with adjacent segment degeneration had significantly less lordosis through the fusion and lumbar spine following their initial surgery. There was no significant difference in the amount of preexisting adjacent level degeneration and smoking between the adjacent segment degeneration and control groups. Fusion of the lumbar spine in abnormal sagittal alignment, with loss of lumbar lordosis, predisposes patients to the development of adjacent segment degeneration. Adjacent segment degeneration does not appear to be just a progression of preexisting degenerative changes at the adjacent level. PMID- 19292355 TI - 11beta-hydroxysteroid dehydrogenase type 1 expression in periprosthetic osteolysis. AB - Periprosthetic membranes contain fibroblasts, macrophages and cytokines including interleukin 1Beta (IL-1Beta) and tumor necrosis factor alpha (TNF-alpha). Glucocorticoids may play an inhibitory role in osteolysis through the upregulation of 11Beta-hydroxysteroid dehydrogenase type 1 (11Beta-HSD1) in membrane fibroblasts by IL-1Beta and TNF-alpha. This study evaluated 15 periprosthetic membranes for the presence of 11Beta-HSD1 using immunochemistry. Also, fibroblast cell cultures were exposed to IL-1Beta and TNF-alpha, and 11Beta HSD1 gene expression was evaluated. In all membranes, 11Beta-HSD1was present, and fibroblast 11Beta-HSD1 was upregulated significantly after the addition of IL 1Beta and TNF-alpha. These findings suggest increased 11Beta-HSD1 expression in fibroblasts may be a mechanism for increasing local cortisol levels in membranes, which could potentiate the osteolytic process through inhibition of osteoblastic function. PMID- 19292356 TI - The accuracy of magnetic resonance imaging in determining the osteotomy plane in osteosarcoma. AB - This study evaluated the accuracy of T1-weighted magnetic resonance imaging (MRI) in determining the osteotomy plane in 21 patients with osteosarcoma undergoing limb-salvage surgery. Twelve cases involved the distal femur and nine cases involved the proximal tibia. Mean patient age was 16.3 years (range, 12-24 years). None of the patients presented with evidence of metastasis. After being placed on neoadjuvant chemotherapy, all patients were treated with en bloc resection and a custom prosthesis. Intramedullary extension was measured on preoperative MRI and radiographs, and also on postoperative specimen by gross and histopathological evaluation. The osteotomy plane was confirmed at 30 mm distal from the primary tumor based on T1-weighted MRI. Mean intramedullary extension measured on MRI, radiographs, and gross examination were 107.4+/-34.5, 78.6+/ 25.6, and 92.6+/-20.5 mm, respectively; actual mean extension was 104.3+/-32.8 mm. There were no significant differences between the actual extension and the extension measured on MRI according to statistical analysis. Intramedullary extension was measured accurately on MRI, which also confirmed the surgical margins. These findings indicate using 30 mm distal from the primary tumor as the osteotomy plane based on T1-weighted MRI is safe. PMID- 19292357 TI - Efficacy of repeat shoulder magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) is frequently used in the assessment of patients with shoulder pain. Occasionally, physicians will obtain a repeat MRI of a shoulder that has not had surgical intervention. The purpose of this study was to determine whether repeat shoulder MRI is diagnostically efficacious. All shoulder MRIs (1252 studies) performed at the University of Texas Medical Branch (UTMB) over a 5-year period were retrospectively reviewed. Patients with a surgical intervention prior to repeat MRI were excluded. A total of 19 patients with 22 repeat shoulder MRIs (1.8% of all shoulder MRIs) were identified. The initial and repeat shoulder MRI findings were compared to determine whether a change in pathology could be detected. In 12 (54%) repeat shoulder MRIs studied, significant progression of pathology was noted. The initial MRI did not depict a rotator cuff tear in 17 patients, while subsequent MRI was remarkable for a cuff tear in 8 (47%) of these patients. Furthermore, of the 12 patients demonstrating considerable progression of disease, 4 (33%) had a repeat shoulder MRI within 90 days. The results of the study demonstrate that MRI is an effective modality for detecting shoulder pathology. In select cases in which the findings may be unremarkable, repeat shoulder MRI may be indicated. PMID- 19292358 TI - The effects of suramin in prevention of peritendinous adhesions following flexor tendon injury in a chicken model. AB - Peritendinous adhesions are the most common complication of flexor tendon injuries. Overproduction of transforming growth factor Beta has been reported as a major cause of tissue fibrosis. This study investigated the effects of suramin, a known inhibitor of transforming growth factor Beta, on the formation of flexor tendon adhesions in a chicken model. Forty-two chicken flexor tendons that were divided partially were repaired using a modified Kessler technique. The chickens were divided into 3 groups. In group 1 (control), no suramin was administered to the tendon repair site; in group 2, 1 mg of suramin was injected around the tendon repair site; and in group 3, 2 mg of suramin was injected around the tendon repair site. At 6 weeks postoperatively, the animals were sacrificed, and functional, histologic, and biomechanical examinations were performed. There was no difference between the groups in terms of tendon excursion. Histologic evaluation showed adhesions were decreased in the groups treated with suramin. In the biomechanical evaluation, a statistically significant difference was present only between the group treated with 2 mg of suramin and the control group. Although suramin is effective in preventing peritendinous adhesions, it can reduce tendon strength as dose-dependent. Future studies should be performed with different doses to determine clinical application. PMID- 19292359 TI - The effect of drill hole size on the fixation strength of a cemented prosthetic patellar button. AB - This study used a cadaver model to analyze the fixation strength of 3 different patellar resurfacing preparations using an all-polyethylene patellar component. One of 3 drill hole sizes was randomly selected and used to prepare the patella. The entire construct was cemented, mounted to a servohydraulic testing machine, cycled between 0 and 50 N, and then tested to failure. Mean forces obtained prior to failure were 258.5, 293, and 353.1 N for the chamfer, 4.5-mm, and 9.5-mm drill holes, respectively. There was a statistically significant difference in strength to failure between the 9.5-mm and chamfer drill hole sizes. These findings may help to reduce patellar implant failures. PMID- 19292363 TI - Lateral subtalar dislocation. AB - Subtalar dislocations are rare in routine orthopedic practice. While many of these dislocations are a result of high-energy injuries such as fall from a height or traffic accidents, it is not uncommon for patients to present after slipping down a few stairs. Two types of dislocation have been described, medial and lateral. The type of dislocation is described according to the position of the foot. In lateral subtalar dislocation the head of talus is found medially and the calcaneus is dislocated laterally. The navicular may lie dorsolateral to the talus. The reverse is true of lateral dislocation. Medial dislocation has been referred to as "basketball foot" due to its preponderance in basketball players.4 The deciding factor is the inverted or everted position of the foot when the force is dissipated through the weak talonavicular and talocalcaneal ligaments. This article presents a case of an adult with lateral subtalar dislocation following a fall. PMID- 19292364 TI - Arthroscopic removal of separated bipartite patella causing snapping knee syndrome. AB - The bipartite patella is a developmental osseous variant that is found in approximately 2% to 6% of the population, and is bilateral in 50%. The proposed etiologies include old nonunion, osteochondritis, and congenital growth defect. It is often found incidentally around the inferior pole, lateral margin, or superolateral border of the patella. It is usually asymptomatic, but may be related to anterior knee pain. However, separation of the bipartite patella is rare, with 9 cases reported in the literature. The symptomatic snapping knee syndrome may be caused by multiple intra-articular and extra-articular pathology including discoid meniscus, tumors, iliotibial band, popliteus, gracilis, semitendinosus, or biceps femoris tendon. However, no reports exist on separated bipartite patella as the feasible cause of the snapping knee syndrome in the orthopedic literature. This article presents a case of snapping knee syndrome due to separated bipartite patella. The accessory bone was removed by arthroscopy, which has rarely been described in the literature. PMID- 19292365 TI - Quadriceps tendon rupture and periosteal sleeve avulsion fracture of the proximal patella pole in an active adolescent. AB - A unique case of anterior knee pain in an active adolescent is described: a periosteal sleeve avulsion and quadriceps rupture in a young basketball player. The trauma mechanism was without any direct trauma on the knee. A review of overuse-related anterior knee pain in the young and active is presented. A quadriceps rupture with a periosteal sleeve avulsion fracture in an active adolescent is very rare. To our knowledge there are no similar cases described. This article presents a case of a 14-year-old adolescent boy who sustained a quadriceps rupture with a periosteal sleeve avulsion. PMID- 19292366 TI - Idiopathic lumbar spinal subdural hematoma. AB - Reports of spinal subdural hematoma are rare. In the few reported cases, type of onset, symptoms, and course have varied, precluding diagnosis based on simple radiography. Obtaining a definitive diagnosis and deciding on a treatment approach can thus sometimes be difficult. Spinal subdural hematoma is classified as acute, subacute or chronic. With acute spinal subdural hematoma, severe and sudden back pain occurs and progression to paraplegia is rapid, within several days. With subacute spinal subdural hematoma, progression to paraplegia occurs slowly, over a period of > or =1 week. Although several cases of spontaneous resolution have been described early surgical treatment is commonly required. This article presents a case of an 85-year-old woman with subacute spinal subdural hematoma who regained the ability to walk following surgical treatment. PMID- 19292367 TI - Low-grade intramedullary osteosarcoma of the tibia presenting as an intracortical lesion. AB - Intramedullary or intraosseous osteosarcomas are the most common subtype of osteosarcoma, followed in frequency by parosteal and periosteal osteosarcomas, with intracortical osteosarcomas representing the rarest type. The diagnosis and classification of osteosarcomas can be complicated or delayed when these lesions masquerade as benign bone lesions. This article presents a case of a low-grade osteosarcoma arising in the tibia of a 19-year-old man, which had radiographic features most suggestive of a benign entity. PMID- 19292368 TI - Hip rotationplasty with antibiotic-loaded bone cement spacer for severe infection following limb-sparing surgery. AB - Knee rotationplasty was initially proposed for the reconstruction of the knee joint in the congenital hypoplasia of the femur. Its application was extended to functional reconstruction of the knee joint after wide resection of malignant bone or soft tissue tumor around the knee. It has also been shown to salvage a failed knee-sparing surgery due to infection or the aseptic loosening of the prosthesis. Hip rotationplasty has been described as a method for the reconstuction of hip function, as well as in the knee joint, in the case of a primary malignant tumor of the proximal part of the femur in children. It has also been described as having a surgical application for the severe congenital deficiency of the proximal part of the femur to reconstruct hip and knee joints, as well as for the massive bone defect of the proximal part of the femur due to infection to mimic a functional femoral shaft. This article reports a case where the hip joint was secondarily reconstructed with hip rotationplasty after subtotal resection of the femur due to infection of the hip hemiarthroplastic prosthesis and osteomyelitis of the hip joint and femur. PMID- 19292369 TI - Hydatid bone disease of the femur. AB - Osseous hydatid disease is caused by the parasitic tapeworm Echinococcus. The species most responsible for hydatid disease is Echinococcus granulosus, endemic especially in sheep-rearing districts like Mediterranean countries and Australia. In Turkey, the exact incidence of human hydatid disease is not known, but < or =34% of asymptomatic farmers have positive serology. Bone hydatidosis is rare, making up 0.5% to 4% of all cases. Patients usually present with pain, swelling, or pathological fracture. There are no specific radiographic signs in affected bone. In the later stages, lytic lesions with a trabeculated pattern, with or without sclerosis, may be seen. Computed tomography (CT) is still the best method for diagnosis and posttherapy follow-up of osseous hydatidosis. On CT, skeletal cystic hydatidosis appears as one or several closely related, well-defined, osteolytic lesions. There may be bone expansion, cortical thinning, cortical destruction, sclerosis, honeycomb appearance, and extension into adjacent soft tissues. PMID- 19292370 TI - Trochanteric excision following persistent nonunion of the greater trochanter. AB - Trochanteric nonunion associated with total hip arthroplasty (THA) may result from 1 of 3 situations: 1) isolated fracture of the greater trochanter; 2) complication of trochanteric osteotomy; or 3) fracture secondary to cystic formation or osteolysis from wear debris post-THA. Isolated fracture of the greater trochanter can occur during primary or revision THA or as a result of trauma in the post-THA patient. It is estimated that approximately 5% of intraoperative femoral fractures involve just the greater trochanter. Perhaps the most common case is subsequent to trochanteric osteotomy. Although trochanteric osteotomy is more often used in revision THA, it may be used in primary THA where there is severe femoral deformity, developmental dysplasia of the hip, or in cases where previous intraosseous hardware is removed. Finally, trochanteric nonunion may be seen in subsequent to late fracture through an osteolytic lesion of the greater trochanter. Regardless of the etiology, nonunion of the greater trochanter can result in significant pain, Trendelenberg gait, and functional limitations, and additional surgery to reattach the trochanter may be required. Reattachment may be accomplished using various techniques, including abductor slide, trochanteric claw plate, Dalls-Miles cable grip system, and various wiring techniques. The nonunion rate following trochanteric osteotomy ranges up to 6%, and persistent nonunion following reattachment ranges up to 39%. PMID- 19292371 TI - Chronic nonhealing ulcer of the right thumb with multiple subcutaneous nodules. AB - A 60-year-old man presented with a 3-month history of nonhealing ulcer over the tip of his right thumb. The ulcer started as a blister over the tip of the thumb that later ruptured and spread proximally to cover the whole pulp area of the thumb. There was no history of trauma, fever, weight loss, or loss of appetite. He is a pensioner and an avid gardener. He has a few cats as pets. The patient initially presented to a private orthopedic surgeon with a nonhealing ulcer of the right thumb. Multiple debridements were unsuccessful in ameliorating the ulcer. Three months after the onset of the initial lesion, multiple painless erythematous nodules had developed on his right arm, and one on the right thigh. All routine blood investigations were nondiagnostic. Swab culture from the ulcer failed to grow any organism and a course of antibiotics did not resolve the problem. Cultures of the biopsy specimen using Sabouraud's dextrose agar and potato carrot medium grew dark brown plaques that microscopically appeared to be branching hyphae. A diagnosis of sporotrichosis of the right upper limb was made and the patient was started on antifungal treatment immediately (T. Itraconazole [Sporanox] 200 mg BD). One month after commencement of antifungal treatment, the ulcer began to dry up and at 3 months all the lesions including the one on the right thigh had healed. PMID- 19292372 TI - Pneumothorax following infraclavicular brachial plexus block for hand surgery. AB - The infraclavicular brachial plexus block is a routinely used anesthesia technique for orthopedic hand, wrist, and arm surgeries. Although the pulmonary anatomy surrounding the brachial plexus would suggest a theoretical potential for pneumothorax development during infraclavicular brachial plexus blockade, this complication is rarely reported in the literature. We report two occurrences of pneumothorax development following routine infraclavicular brachial plexus blockade performed by physicians at an academic training institution with technical experience. Both patients were healthy adult males with a normal body habitus undergoing elective upper extremity surgical intervention. They presented to the emergency department on postoperative days 1 and 2 with shortness of breath and chest pain. Despite different clinical management strategies for each patient, including conservative physical activity with inspirational spirometry compared to thoracic decompression, both patients' pneumothoraces resolved with no apparent sequelae. This article describes the clinical benefits of infraclavicular brachial plexus blockade versus general anesthesia during elective outpatient surgeries. The development of symptomatic pneumothoraces in 2 healthy adult patients stresses the importance of careful patient assessment pre- and postoperatively. Possible preventative strategies to avoid pneumothorax complications during infraclavicular brachial plexus blockade are discussed. This case report indicates there is a risk of iatrogenic injury during infraclavicular brachial plexus blockade. This information could be valuable in determining anesthesia modalities used for orthopedic upper extremity surgeries in patient populations with problematic follow-up or limited access to health care. PMID- 19292373 TI - Hypersensitivity to polymethylmethacrylate following shoulder hemiarthroplasty. AB - Acrylic resins have been used for many years in several health-related applications due to their ease of use, favorable material properties, and relative cost. Cements containing polymethylmethacrylate (PMMA), in particular, have been widely accepted for use in orthopedic surgery, as well as in other fields of medicine. Although relatively rare, the potential for acrylic resins such as PMMA to induce hypersensitivity reactions via cutaneous or mucosal exposures has been reported; however, comparatively few cases have been described of patients reacting adversely to acrylic resins used as permanent cements during surgical procedures. This article reports a hypersensitivity reaction to PMMA cement applied in a right shoulder hemiarthroplasty, which initially presented as a possible postoperative infection. It is believed to be the first case in the literature of such a reaction occurring in an upper extremity prosthesis. PMID- 19292382 TI - Comparison of bony healing and cartilage degeneration following strut grafting and trapdoor procedures in an ovine model. AB - This experimental study compared bony healing and cartilage degeneration following strut grafting and trapdoor procedures for osteonecrosis of the femoral head in an ovine model. Osteonecrosis and a bony defect were created surgically in 16 hips of 8 Merino sheep, and the index grafting procedures were performed in a second session. Three months after surgery, the hips treated by strut grafting yielded better articular cartilage protection and bony healing compared to the osteonecrosis groups. The trapdoor group yielded better bony healing compared to the rest of the groups, but the grafts and the osteochondral flaps failed to unite with the host bone. These findings indicate elevating the osteochondral flap during the trapdoor procedure injures the cartilage and may cause degeneration. PMID- 19292383 TI - Arthroscopic capsular shrinkage for chronic ankle instability with thermal radiofrequency: prospective multicenter trial. AB - The study was designed as a prospective multicenter longitudinal trial. Adult patients with symptomatic mechanical chronic ankle instability, not improving with conservative therapy, were included and underwent surgery. Primary outcome measures were radiological and manually tested mechanical laxity. Secondary outcome measures were number of complications, reoperations and symptoms, range of motion, and functional (ankle) scores (Karlsson and SF-36 score). The latest follow-up was 9 months for each patient. Thirty-nine patients underwent surgery (19 male patients; 16 right ankles; median age, 27 years). Mechanical stability showed no clinically relevant improvement whereas most secondary outcome measures showed a substantial and statistically significant improvement. One surgery related complication occurred without functional consequences, and 3 patients underwent a secondary procedure. One was considered a treatment failure, requiring an open anatomic ligament reconstruction. The second patient sustained a severe supination trauma by starting intensive training too early, also requiring an open anatomic reconstruction. The third patient had posterior ankle pain, which was successfully treated by posterior ankle arthroscopy. Arthroscopic thermal capsular shrinkage of the ankle is a safe procedure, leading to resolution of symptoms in the majority of patients with chronic ankle instability. PMID- 19292384 TI - Consistent new bone formation in 95 revisions: average 9-year follow-up. AB - This study evaluates the use of Wagner cementless stem for femurs with marked loss of proximal bone stock. Ninety-five hips in 89 patients were revised with a Wagner stem and prospectively studied. The indications were as follows: aseptic loosening for 84 hips, periprosthetic fracture for 5, Girdlestone for 3, mechanical loosening for 2 and recurrent dislocation for 1. The mean follow-up was 108 months (range, 84-180 months; SD, 13.5). There was only 1 re-revision, 2 months after surgery, and the survival rate at 9 years was 98.5%. The mean Merle d'Aubigne score improved from 8.8 to 16.1 (P<0.001). There were 15 (15.8%) intraoperative fractures, 5 (5.3%) dislocations, 2 (2.1%) thrombosis, and 2 (2.1%) sciatic neuropraxia. Significant bone formation occurred around the Wagner stem in 97% of the hips in these cases. No radiolucency or loosening had been diagnosed at the last follow-up. PMID- 19292385 TI - Fracture load for periprosthetic femoral fractures in cemented versus uncemented hip stems: an experimental in vitro study. AB - This cadaveric study examined fracture loads in cemented and uncemented hip stems. Additionally, individual data and bone quality were analyzed and correlated to fracture patterns and fracture load. Cemented or uncemented hip stems were implanted in a randomized fashion in 10 matched paired fresh-frozen femora (donor median age, 78 years, and donor median weight, 74.2 kg). Bone density was measured before the femurs were fractured under load (maximum load of 10,000 N), and fracture patterns were analyzed according to the Vancouver and Johansson classification systems. In the uncemented group, all of the femurs fractured with a median load of 2625 N (range, 1725-7647 N). In the cemented group, 5 femurs fractured with a median maximum load of 9127 N (range, 2845 10,000 N) and 5 femurs did not fracture with a maximum load of 10,000 N. Fracture load corresponded to 4 times and 8.8 times body weight in the uncemented and cemented groups, respectively. Fracture patterns corresponded to Vancouver type A fractures in uncemented stems and Vancouver type C fractures in cemented hip stems. Analysis showed a significant correlation between fracture load and bone density in the uncemented group, whereas there was no correlation in the cemented group. Patients with poor bone quality treated with an uncemented hip stem are at higher risk for periprosthetic fractures; therefore, we recommend cemented stems in this group of patients. Cementation appears to protect against periprosthetic fractures, probably from internal stiffening of the femoral cavity. PMID- 19292386 TI - Gender differences in the distal femur dimensions and variation patterns in relation to TKA component sizing. AB - Matching femoral components with distal femurs in female patients can be difficult. The purpose of this study was to determine the validity of the following hypotheses: (1) Anteroposterior (AP) dimensions do not increase proportionately to medial-lateral (ML) dimensions in men and women. (2) The AP/ML ratio is different in men and women. (3) The AP/ML ratios of femoral implants and the variations in ratios are more in line with data from men than women. Magnetic resonance imaging of the knees of 50 consecutive male patients and 50 consecutive female patients were studied. Both groups had similar age distribution. Anteroposterior and ML dimensions were measured, and AP/ML ratios were obtained and plotted. Inside AP and ML dimensions of 8 popular total knee arthroplasty (TKA) systems produced by 6 manufacturers were obtained. The AP/ML ratio for women was 0.80 (SD=.05), whereas that for men was 0.76 (SD=.03). This study demonstrated a significant difference in distal femur proportions between men and women. Dimensions propagation of femoral components for TKA followed significantly closer to men's distal femur dimensions variation than to that of women. PMID- 19292387 TI - Acute aortic dissection or ruptured aortic aneurysm associated with back pain and paraplegia. AB - Acute aortic dissection and a ruptured aortic aneurysm are both catastrophic events that usually present suddenly. Although these conditions are relatively uncommon compared to other diseases treated by the orthopedists as a primary practitioner, they are of primary importance. The purpose of this study was to investigate the key points that differentiate these conditions from spinal disease. A review of 50 patients with aortic dissection (n=40) or a ruptured aortic aneurysm (n=10) was performed to determine the manifestations (eg, back pain and paraplegia). We also evaluated the predisposing factor and blood pressure on admission and reviewed clinical imaging (radiographs, computed tomography [CT]) retrospectively. Sudden severe isolated back pain was observed in 18 (45.0%) of 40 patients, and 31 (77.5%) of 40 patients had at least some back pain in aortic dissection, while 1 patient had sudden paraplegia with a ruptured aortic aneurysm. Hypertension was the most predisposing factor and was present in 29 (58.0%) of 50 patients. On admission, hypertension was present in 26 (56.5%) of 46 patients, and hypotension was present in 14 (30.4%) of 46 patients. In all cases, the correct diagnosis was made based on CT. For a patient with an abrupt onset of severe back pain, acute aortic dissection and a ruptured aortic aneurysm should be considered in the differential diagnosis from spinal disease. The most reliable tool for imaging diagnosis was CT. PMID- 19292388 TI - Trends in surgical management for type II odontoid fracture: 20 years of experience at a regional spinal cord injury center. AB - This study reviewed surgical treatment methods in patients with type II odontoid fracture who were admitted to a level 1 spinal cord injury center during a 20 year period. Of the 186 patients who met inclusion criteria, 75 were treated operatively. The rate of surgical intervention increased during the study period. Length of hospital stay was not related to treatment. Estimated blood loss was significantly higher for posterior techniques. The Brooks technique and anterior odontoid screw fixation declined in frequency, whereas the Magerl technique has been used at a consistent rate since its introduction. The C1 lateral mass and C2 pedicle/isthmus screw technique has increased significantly since its description. Pneumonia, vocal cord, and swallowing problems occurred more often with anterior approaches. PMID- 19292389 TI - Efficacy of composite allograft and demineralized bone matrix graft in treating tibial plateau fractures with bone loss. AB - Tibial plateau fractures with bone loss or significant comminution require grafting and stable fixation. We hypothesized a standardized protocol of internal fixation augmented with a mixture of demineralized bone matrix and corticocancellous allograft chips would result in high healing rates with minimal subsidence. Union was achieved in all 36 patients available for follow-up by a mean of 4.4 months. Mean range of motion was 2 degrees to 120 degrees. One patient developed osteomyelitis. Subsidence ranging from 2.5 to 5.7 mm occurred in 4 patients (11%). This treatment method provides sufficient structural integrity with a high union rate and a low complication rate. PMID- 19292390 TI - Effect of olecranon resection on joint stability and strain of the medial ulnar collateral ligament. AB - This study aimed to assess the effect of posteromedial olecranon resection on the stability of the elbow joint, forearm kinematics, and strain in the anterior bundle of the medial ulnar collateral ligament. Seven elbows were tested after resecting all soft tissues except the capsuloligamentous structures. Varus-valgus angulation, forearm rotation, and the length of the anterior bundle of the medial ulnar collateral ligament were measured at 3 posteromedial olecranon resection stages of 0, 4, and 8 mm, and at elbow flexion angles of 30 degrees, 60 degrees, and 90 degrees. Posteromedial olecranon resection significantly increased varus and valgus laxities at 60 degrees and 90 degrees of elbow flexion but had a larger effect on varus motion than valgus movement. With more posteromedial olecranon resection, the valgus angulation and varus-valgus laxity proportionally increased. In most cases, parallel resections of the posteromedial olecranon significantly decreased the functional varus-valgus stiffness. There was no significant effect on strain in the anterior bundle of the medial ulnar collateral ligament due to olecranon resection. When treating an overhead athlete with posteromedial olecranon impingement, the resection should be limited to osteophytic overgrowth. Throwers may be susceptible to ulnar collateral ligament injury following posteromedial olecranon resection due to increased elbow laxity. PMID- 19292391 TI - Determinants of patient satisfaction following surgery for multidirectional instability. AB - Fifty shoulders in 46 patients underwent stabilization surgery for multidirectional instability. Univariate analysis showed no significant differences (P>0.05) for age, gender, or workers' compensation in patient satisfaction or American Shoulder and Elbow Surgeons (ASES) score. Only the ASES score was lower with prior surgery (P=0.001). There was a significantly increased ASES score (P<0.05) for arthroscopic versus open treatment. Subjective variable analysis showed that satisfaction and ASES score were significantly associated with questions regarding pain, instability, and upper extremity use (P<0.01). Increased range of motion showed a trend toward higher ASES scores (P<0.074). Patient satisfaction (P=0.013) was associated with greater forward elevation and greater external rotation (P=0.056). Multivariate analysis (P<0.05) showed that independent determinants of patient satisfaction with outcome included change in instability symptoms and ASES score. Subjective variables of symptoms and motion had the greatest correlation with patient satisfaction and ASES score following surgery for multidirectional shoulder instability. To improve patient satisfaction, an expanded focus on these subjective points may be beneficial. PMID- 19292392 TI - Effectiveness of interference screw fixation in ulnar collateral ligament reconstruction. AB - Ulnar collateral ligament disruptions of the elbow are increasingly common for athletes involved in overhead sports. One newer reconstructive technique combines the use of a biotenodesis screw for ulnar fixation with the docking procedure for humeral fixation referred to as the DANE procedure. Biomechanical evaluations have determined that the combined procedure has properties similar to previous methods, but few comparative clinical studies have been performed. The current study compares the DANE procedure to a traditional method of ulnar collateral ligament reconstruction with a medium range follow-up. Twelve ulnar collateral ligament reconstructions were observed for a minimum of 21 months. Six reconstructions were performed using bone tunnels on the ulna. The other 6 used a biotenodesis interference screw on the ulna. Both groups used the docking procedure on the humerus. Identical rehabilitation phases were followed by both groups. Attention was paid not only to the time to return to play and the level able to be returned to but also to interoperative differences, including tourniquet time. We concluded that the DANE reconstruction is an appropriate and effective method. Results are similar to those achieved with a traditional reconstructive method. The study also demonstrates that tourniquet time and, therefore, operative time is less for the DANE procedure. PMID- 19292395 TI - Radiologic case study. Cervical spine trauma: unstable fractures, C2-C7 injuries, and imaging guidelines. AB - Enhance your diagnostic skills with this "test yourself" monthly column, which features a radiograph and challenges you to make a diagnosis. PMID- 19292402 TI - Multislice helical CT in the diagnosis of pharyngovertebral fistula due to loosening of anterior screw-plate construct. AB - Late dysphagia is a rare condition that results from hardware failure in anterior spine surgery patients. Dysphagia should be evaluated to find out the possible underlying causes. A barium swallow study and multislice helical computed tomography (CT) scan were used to evaluate a patient suffering from dysphagia 8 months after an anterior cervical fusion of C4-C7 vertebrae. A videofluoroscopic swallow study with a low-viscosity, high-density barium solution and different table tilt levels for obtaining slow passage of the barium meal revealed the existence of an esophagovertebral fistula with 2 ostia: one at the contact side of the screw and the other at a lower level with a pouch around the plate. A multislice helical CT scan provided volumetric data and multiplanar reconstruction that enabled us to delineate the extent of the abscess along the plate through the paravertebral space. It also showed the fistula location and relation of the fistula to the displaced screw, as well as the thickened retroesophageal soft tissue. Anterior spinal surgery patients should be followed carefully for progressive dysphagia. Videofluoroscopic swallow studies and multislice helical CT scans are efficient and complementary tools in the evaluation of late dysphagia. PMID- 19292403 TI - Delayed symptoms of cerebrospinal fluid leak following lumbar decompression. AB - Numerous reports in the literature describe cerebrospinal fluid (CSF) leaks resulting from dural tears; however, most of these reports document symptoms of incidental durotomy occurring ,48 hours postoperatively. This case report describes patients with symptoms of CSF leak occurring a few days to a few weeks after lumbar spine surgery who required additional surgery to repair the dura and alleviate their symptoms. Records for patients who had lumbar spine surgery performed by 2 spine surgeons at a single institution between 1990 and 2005 were reviewed. After additional surgery to repair what appeared to be late postoperative durotomies, 2 patients recovered fully from their orthostatic headaches and reported satisfactory relief of their preoperative neurological symptoms. They did not have symptoms of spinal fluid leakage following surgery, and no intraoperative dural tear was noted, suggesting these cases may have been secondary to late postoperative durotomy. The incidence of durotomy has been noted to be increased in patients undergoing revision spine surgery secondary to adhesions and scarring of the dura. As this was the first spine operation for both cases reported, we believe that residual bone spikes are responsible for puncturing the dural sac postoperatively. Spine surgeons should be exceedingly cautious in inspecting for bone spikes following an extensive dural exposure and recognize the significance of the new onset of an orthostatic headache, even days or weeks following spine surgery. PMID- 19292404 TI - Compartment syndrome following intraosseous infusion. AB - Intraosseous infusion is a valuable technique in the resuscitation of critically ill pediatric patients in whom vascular access has proved otherwise impossible. Although it is well established as a safe and reliable means of emergent access, intraosseous infusion is not without danger, nor complication. One of the rare yet most grave complications of intraosseous access is compartment syndrome. We report a case of compartment syndrome as a result of intraosseous infusion that serves to remind of the potential pitfalls of this technique. An otherwise healthy 6-year-old girl presented to our institution's pediatric intensive care unit following emergent resuscitation for a prolonged cardiac arrest. Approximately 1 hour following an uneventful soccer practice, without any antecedent cardiopulmonary symptoms or complaints, the patient collapsed and was unresponsive, not breathing, and pulseless. In the course of resuscitation, right and left tibial intraosseous lines were started. After 30 minutes of resuscitation, with multiple rounds of lidocaine and epinephrine infused through the intraosseous lines, a sustained perfusing rhythm was established. Acute compartment syndrome was diagnosed, and through anterolateral and posteromedial incisions, all 4 fascial compartments were released. While the condition of the patient's extremity improved, the overall clinical condition of the patient did not. This case highlights the fundamental principles regarding the use of intraosseous infusion and the diagnosis and management of compartment syndrome in critically ill patients. PMID- 19292405 TI - Postsurgical bleeding following treatment of a chronic Morel-Lavallee lesion. AB - Morel-Lavallee lesions are closed soft-tissue degloving injuries that occur when the skin and subcutaneous tissues are separated from underlying fascia as a result of a sudden shearing force. The space that is created has the potential to be filled with a mixture of blood, lymph fluid, or necrotic fat, which may easily become infected. The optimal treatment of Morel-Lavallee lesions is controversial. We present a case of a 29-year-old man who sustained pelvic fractures in a motorcycle accident and 1 year later presented with a painless, slowly expanding left thigh mass. Treatment, consisting of open debridement and primary closure, was complicated by significant postoperative bleeding requiring reoperation. The patient was followed clinically as an outpatient, and at 6-month follow-up he was doing well and had no evidence of recurrence of the lesion. Morel-Lavallee lesions are rare complications that are often missed or underappreciated at the time of initial injury. In many cases these lesions can persist for months before they are recognized. For long-standing Morel-Lavallee lesions, it is important to determine the nature of the fluid in the cavity before planning treatment. In the current case, it was unclear based on the official radiographic interpretation whether the lesion was filled with frank blood versus serous or serosanguineous fluid. PMID- 19292406 TI - Disassembly of cemented bipolar prothesis of the hip. AB - A case of spontaneous disassembly of a bipolar prothesis after hemiarthroplasty due to a dislocation of the hip in an 85-year-old woman is presented, which is a rare complication. Open reduction was performed and original components were used because there was no acetabular and polyethylene linear erosion. The result was satisfactory at final follow-up (36 months), and radiographs showed no acetabular erosion, polyethylene wear, femoral subsidence, or component migration. To date, only 6 cases of disassembly of bipolar components have been reported in the English literature. In almost all cases, the injury was caused without any significant force, as only 2 patients reported a fall. Most were elderly women. We classified them into 3 types according to the etiology: iatrogenic disassembly (type I), erosion of the polyethylene liner (type II), and spontaneous disassembly (type III). Spontaneous disassembly is uncommon and seemed to be caused by a failure of the locking mechanism between the head components, resulting in an intraacetabular dislocation. Open reduction is required and revision surgery is judged by the locking mechanism between components. When there is no acetabular and polyethylene linear erosion, we believe that the original components can be used again. Although the potential disadvantage of bipolar systems shown here does not outweigh the advantages, we should rethink the status of bipolar hemiarthroplasty. PMID- 19292407 TI - Protrusion of the ceramic head through the acetabular shell in total hip replacement. AB - Total hip replacement (THR) is one of the most successful elective operations currently being performed. However, increased implant wear in the femoral head acetabulum articulation has been found to limit the long-term survivorship of the procedure. Clinical complications of this wear process include mechanical failure of the polyethylene insert as well as a biologic foreign-body response to wear debris. Factors that have been implicated in the pathogenesis of this phenomenon include implant design and positioning, patient age, and time from the operation. Pain and sound originating from the joint are among the most common clinical manifestations. Radiographic signs present in the form of progressive radiolucent zones or an eccentrically displaced femoral head. The true extent of bone damage usually is greater than that appreciated originally. A high index of suspicion is essential for an early and successful surgical intervention. We report a rare case of a 72-year-old woman who presented with excessive wear of both the acetabular polyethylene socket and metal backing 3 years after the initial cementless THR. This resulted in protrusion of the femoral head through both the polyethylene cup and the acetabular shell, and subsequent subluxation of the femoral head. Marked periacetabular osteolysis and loosening had developed progressively. No definite signs of infection were identified, either by bone scintigraphy, preoperatively, or by repeated cultures of the excised tissues. The THR was revised, in a 2-stage operation, using bone autografts and a cemented acetabular socket, with a good postoperative outcome. PMID- 19292408 TI - Synovial chondromatosis of the radiocarpal joint. AB - Synovial chondromatosis is an uncommon lesion of unknown etiology, characterized by progressive metaplasia associated with the formation of cartilage in the synovial membranes of joints, tendon sheaths, or bursae. Synovial chondromatosis is typically monoarticular and affects large joints such as the knee and hip, although it has also been described in the ankle, elbow, and shoulder. However, it rarely occurs in the wrist. Our literature review showed only 28 cases with wrist involvement, 13 cases with distal radioulnar joint involvement, 12 cases with midcarpal joint involvement, and 3 cases with radiocarpal joint involvement. This article presents a case of synovial chondromatosis of the radiocarpal joint. A 45-year-old right-handed woman presented with a 4-year history of a palpable mass and pain in her right wrist. On physical examination, 2 masses were palpable on the volar and radial aspect of the joint. Plain radiographs of the right wrist demonstrated well-defined radiopaque calcified mass lesions. Cortex and subcortical bone on the radial aspects of the radius and scaphoid were eroded. T1 weighted magnetic resonance imaging (MRI) showed a low-signal-intensity lesion, and T2-weighted MRI showed a heterogeneous high-signal-intensity lesion in the radiocarpal joint, which was consistent with synovial chondromatosis. Operatively, 2 glistening white lobulated masses were removed together with attached synovia. A histologic examination confirmed synovial chondromatosis with dystrophic calcification. At the time of the most recent examination (3 years postoperatively), the patient was pain-free, had no residual motion deficit, and showed no radiographic evidence of recurrent disease. PMID- 19292409 TI - An unusual case of olecranon tuberculosis. AB - A 67-year-old woman presented with erythema and swelling over her left elbow. She felt pain during palpation of the olecranon, while the range of elbow motion was slightly reduced (02 in circle-302 in circle-1302 in circle). She had been treated conservatively elsewhere for 3 months with nonsteroidal anti-inflammatory drugs and splint immobilization. Radiographs and computed tomography showed an osteolytic area over the olecranon with cortex disruption. She underwent open biopsy of the olecranon, and the histologic features were compatible with tuberculosis. Erythrocyte sedimentation rate was elevated and she had a positive Mantoux test (14 mm). She mentioned no history of pulmonary disease and had a normal chest radiograph. She received antituberculous treatment (isoniazid, rifambin, and pyrazinamide) for 2 months without obvious improvement. She underwent surgical debridement of the olecranon while she continued antituberculous treatment for 12 months, discontinuing pyrazinamide after the third month. After the completion of antituberculous treatment, the patient was pain-free, without local signs of infection and with improved range of motion (02 in circle-202 in circle-1452 in circle). Early diagnosis and treatment of olecranon tuberculosis is of great value, and the prognosis is good when there is no spread in the elbow joint. PMID- 19292410 TI - Arthroscopic treatment of septic arthritis of the shoulder in a 6-year-old boy. AB - Septic arthritis of the shoulder in children is a rare condition. The diagnosis may present some difficulties and, consequently, appropriate treatment often is delayed. Main sequelae are humeral shortening, joint instability, premature arthritis and limited range of motion. We report a case of septic arthritis of the shoulder in a child who was treated by means of shoulder arthroscopy. A 6 year-old boy presented with a history of fever, pain and functional impairment of the shoulder that were lasting despite having undergone antibiotic therapy for 28 days (amoxicillin per os, and then teicoplanin intravenously combined with meropenem intravenously) and an arthrocentesis (no organisms were identified) in another hospital. Clinical examination (pain, swelling, warmness, functional impairment), laboratory tests (white blood cell count, 6.900/mm(3); C-reactive protein, 6.44 mg/dL; erythrocyte sedimentation rate, 119 mm), and imaging studies (radiographs, ultrasonography, computed tomography scan, magnetic resonance imaging, bone scan) performed in our department suggested the diagnosis of a stage IV (with osseous involvement) septic arthritis. Arthroscopic irrigation, debridement, synoviectomy and shaving of the osteochondral erosions were performed, in association with antibiotic therapy (teicoplanin and ceftriaxone disodium intravenously, and then amoxicillin/clavulanate per os). At 22-month follow-up, the patient was asymptomatic and showed a full range of motion. No limb length-discrepancy was found. Radiographs showed irregular profile of the humeral epiphysis without any physeal disturbances. Arthroscopic treatment for septic arthritis of the shoulder in children, though rarely reported, represents an adequate procedure for cases without bone involvement and may lead to good results, even in stage IV cases. Open arthrotomy should be reserved for cases with concomitant osseous infection after failure of arthroscopic treatment. PMID- 19292411 TI - Telangiectatic osteosarcoma of the patella. AB - Although osteosarcoma is the most common primary malignancy of bone, it has only been reported to arise from the patella in a handful of cases. Telangiectatic osteosarcoma accounts for <5% of all osteosarcomas, and it is distinguished histologically by spaces, often blood filled, separated by septa containing highly malignant cells and radiographically by a predominately lytic and/or expansile component. Telangiectatic osteosarcoma can be radiologically confused with aneurysmal bone cyst or giant cell tumor. A 22-year-old otherwise healthy man presented with increasing pain, swelling, and limited flexion of the right knee after failing physical therapy for anterior knee pain. Standard anteroposterior and lateral radiographs demonstrated a diffuse destructive process involving the majority of the patella (including loss of the inferior patellar cortex) and a lytic lesion of the proximal tibia. Apparent osteoid matrix was visible in the soft tissue extension along the inferior pole of the patella. A computed tomography scan of the chest showed 2 pulmonary nodules consistent with metastatic disease. Evaluation of core needle biopsy showed osteosarcoma with telangiectatic features. Given that the majority of the tumor involved the patella/extensor mechanism, it was clear that the tumor originated in the patella. This case presents the first published report of a telangiectatic osteosarcoma arising from the patella. PMID- 19292412 TI - Wound necrosis after total knee arthroplasty. AB - Quickly evolutive skin necrosis and deep infection after total knee arthroplasty (TKA) are not uncommon. Several predisposing factors, such as immunosuppression, malnutrition, steroid use, rheumatoid arthritis, multiple scars, and vascular disease can be involved in the onset of wound complications, as well as long tourniquet time and early knee flexion. Skin necrosis after TKA can be treated in different ways, including local wound care, debridement, and soft tissue coverage with muscle or skin grafts. This article presents a rare case of skin necrosis occurring in a patient without any other apparent risk factor after TKA. A 78 year-old woman affected by primary osteoarthritis of the right knee who had no comorbidities and who had already undergone TKA for primary osteoarthritis on the left knee underwent a cemented TKA. Three days postoperatively, she developed a fever and wound problems, which soon after turned into skin necrosis. This complication was first treated surgically with a debridement of the wound with antibiotic therapy and local wound care, then with vacuum-assisted closure (Kinetic Concepts Inc, San Antonio, Texas) therapy and soft tissue coverage using skin grafting. She had a complete recovery in the next 3 months; the skin grafting was well tolerated and the range of motion and functional outcome were good. PMID- 19292413 TI - An unusual case of elbow dislocation. AB - The annual incidence of elbow dislocation is 6 to 8 cases per 100,000 in the United States; these dislocations represent 11% to 28% of all elbow injuries. The frequency of elbow dislocation is second to that of shoulder dislocation. Adult elbow dislocations are classified by the direction of displacement and associated fractures. They can be anterior, posterior, lateral, or divergent, with the most common type being posterior displacement of both the radius and ulna in relation to the distal humerus. This case report presents a rare case of a twisting dislocation of the elbow joint without any accompanying fracture. A 55-year-old woman presented with a swollen and painful right elbow joint and a pronated forearm after falling on her outstretched right arm. Closed reduction was performed under local anesthetic. No neurovascular deficit was recorded before or after reduction, and no ectopic calcification was observed at 1 year follow-up. Early mobilization of the elbow resulted in a stable joint with full range of motion. The rehabilitation of elbow dislocation ranges from aggressive immediate active motion to traditional plaster of Paris immobilization for several days. Forceful passive mobilization in the rehabilitation period must be avoided, since the elbow joint has a natural tendency to develop myositis ossificans following passive manipulation. PMID- 19292414 TI - Osteochondral flap avulsion fracture in a child with forearm compartment syndrome. AB - Traumatic elbow dislocations account for approximately 3% to 6% of childhood upper extremity injuries, and 41% of these dislocations are concomitant injuries around the elbow. Most of these injuries are associated with a fall on an outstretched hand. Compartment syndrome is one recognized complication after elbow dislocation and distal humeral and radial neck fracture. Compartment syndrome in the forearm is a devastating complication in children with forearm, elbow, and supracondylar fractures. Compartment syndrome occurs as the result of hypoxic damage caused by interruption of the circulation to the muscles. Any evidence of compartment syndrome requires compartment pressure measurements and possibly fasciotomy. Fasciotomy is recommended in the presence of clinical signs of compartment syndrome, such as undue pain (out of proportion to severity of injury), pallor, paresthesia, absent or deficient pulse, and paralysis of the limb. Osteochondral flap avulsion fracture is a rare clinical presentation in pediatric elbow injuries. To our knowledge only 8 cases have been reported in the literature. Our case is different from others in terms of delayed presentation, and is associated with olecranon fracture and forearm compartment syndrome. This case reemphasizes the anatomy of the semilunar notch of the proximal ulna and the importance of careful clinical and radiological examination in the treatment of childhood elbow injuries. PMID- 19292415 TI - Fracture-dissociation of ceramic liner. AB - The use of BIOLOX delta ceramic (CeramTec AG, Plochingen, Germany) has been increasing. This ceramic prevents cracking by restraining the phase transformation due to the insertion of nano-sized, yttria-stabilized tetragonal zirconia into the alumina matrix. This restrains the progress of cracking through the formation of platelet-like crystal or whiskers due to the addition of an oxide additive. We observed a case of BIOLOX delta ceramic liner (CeramTec AG) rim fracture 4 months postoperatively. Radiographs showed that the ceramic liner was subluxated from the acetabular cup. Scratches on the acetabular cup and femoral neck were seen, and the fracture was visible on the rim of the liner. Under electron microscope, metal particle coatings from the ceramic liner were identified. The ceramic liner, fracture fragments, and adjacent tissues were removed and replaced with a ceramic liner and femoral head of the same size and design. We believe the mechanism of the fracture-dissociation of the ceramic liner in this case is similar to a case of separation of the ceramic liner from the polyethylene shell in a sandwich-type ceramic-ceramic joint. To prevent ceramic liner fracture-dissociation, the diameter of the femoral neck needs to be decreased in a new design, while the diameter of the femoral head needs to be increased to ensure an increase in range of motion. PMID- 19292416 TI - Bilateral femoral supracondylar stress fractures in a cross country runner. AB - Several high-risk factors lead to stress fractures. They include excessive training in athletes leading to overuse injuries, nutritional deficiencies, and endocrine disorders. While stress fractures are common, bilateral stress fractures are rarely seen. Few cases have been reported of bilateral femoral stress fractures in young athletes. This article presents a case of a 14-year-old cross country runner with a bilateral femoral supracondylar stress fracture. He presented with bilateral supracondylar stress fractures from running. The patient followed a strict vegan diet, but his parents stated that, to their knowledge, he was getting adequate protein and calcium. Treatment consisted of decreased activity to pain-free levels with acetaminophen for pain. Low-impact conditioning such as swimming and bicycling was allowed. Hamstring and quadricep stretching was suggested. Nutritional consultation was obtained to ensure appropriate nutrition on a vegan diet. At 1-month follow-up, he was pain free and allowed to proceed with a gradual return to running activities. In this case, the onset of a new workout routine was intolerable for this patient's low bone density, causing insufficiency fractures. Appropriate vegan diets were not associated with stress fracture in our literature review. He may have had an inadequate diet prior to this injury. As in this case, full recovery can be made after this rest period, and the patient may return to his or her original activity safely. In young athletes, diet and nutrition must be kept in mind. PMID- 19292417 TI - Proximal femoral stress reaction in a professional football player. AB - There have been no previous case reports of femoral stress fractures in professional football players. We present a professional defensive back who presented 1 week after noting mild soreness in the right proximal thigh during practices, prior to a regular season game. Early diagnosis of a stress reaction was made with magnetic resonance imaging (MRI). Magnetic resonance imaging is an excellent imaging modality for stress reactions/fractures given its high sensitivity, specificity, and ability to image surrounding soft tissues. Short tau inversion recovery and fat-suppressed images are best for seeing early osseous edema. Early detection of this injury allowed us to hasten the recovery and prevent further injury. The patient was treated successfully with early nonweight bearing, a gradual increase in activity via cross training, and a structured functional progression to return to play. Normalization of MRI lagged behind the player's healing and was not necessary to determine advancement of rehabilitation or return to play. Three main issues are important to review in this case of a proximal femoral stress injury: early diagnosis is the key to treatment and early return to play; imaging studies lag behind clinical symptoms; and the treatment regimen should consist of a period of rest and be tailored to the individual. PMID- 19292419 TI - Sporting activity following discectomy for lumbar disc herniation. AB - The aim of this study was to investigate to what extent patients could resume physical activity following surgery for herniated lumbar disks. We analyzed a cohort of 1003 patients who underwent lumbar spine surgery within 1 year. Out of this cohort, 93 patients were selected according to our inclusion criteria (age 20-35 years, mediolateral single level disk herniation, no comorbidity at the lumbar spine, and treatment with conventional subtotal diskectomy). This group was evaluated after a minimum follow-up of 28 months in a telephone questionnaire; participants were questioned about pre- and postoperative physical activities. The questionnaire was answered by 67 patients. Twenty-six patients were lost to follow-up because they had relocated. The follow-up group had a mean age of 30 years. Five patients underwent a second procedure due to recurrent disk herniation. All patients showed a pain reduction. At follow-up, no patient needed constant pain medication. Eighty-two percent of the patients were pain free during practicing sports. Sixty-two patients performed some type of sport after surgery. Concerning the type and frequency of physical activities, no significant change between pre- and postoperative behavior occurred. The 5 patients with recurrent disk herniation did not behave differently. Single-level lumbar disk surgery does not limit or compromise sportive activity in young people. PMID- 19292420 TI - Course of avascular necrosis of femoral head without collapse of femoral head at first examination: minimum 8-year follow-up. AB - Disruption of the femoral head contour by collapse of necrotic segments is considered to be an important turning point in the history of avascular necrosis of the femoral head, and in many cases, surgical treatment is required soon after the onset of symptoms. However, in a few cases, conservative treatment is sufficiently effective to avoid surgical procedures for long periods. For patients with early-stage avascular necrosis, it is important to accurately predict its natural course. This article documents the characteristics of patients who can be treated without surgery for long periods. Seventeen hips in 13 patients with avascular necrosis of the femoral head without collapse of the femoral head at first examination were followed for at least 8 years of conservative treatment. Long-term outcomes of hips with avascular necrosis were divided into 3 groups: (1) hips without collapse, (2) hips with progression but cessation of collapse, and (3) hips with progression of collapse. In groups 1 and 2, good clinical results are expected. Hips with femoral head collapse .3 mm at 3 years from the onset of hip pain progressed to osteoarthritis. Our analysis indicated that collapse of the femoral head does not necessarily indicate a poor prognosis, and even after collapse occurs, subsequent cessation of collapse can be expected in a certain percentage of hips. PMID- 19292421 TI - The outcome of composite bone graft substitute used to treat cavitary bone defects. AB - Although autogenous bone graft remains the gold standard graft material, it is associated with an unacceptably high incidence of morbidity. Furthermore, operative time, blood loss, and length of hospitalization are often increased. In order for a graft substitute to replicate the optimal bone healing properties of autogenous graft, 3 essential elements must be present: scaffolding for osteoconduction, growth factors for osteoinduction, and progenitor cells for osteogenesis. A composite graft that combines a synthetic scaffold with osteoprogenitor cells from bone marrow aspirate (BMA) may potentially deliver the advantages of autogenous bone grafts without the procurement morbidity. Sixty consecutive patients with cavitary bone defects were treated with a composite of b-tricalcium phosphate (beta-TCP), Vitoss (Orthovita, Malvern, Pennsylvania), and BMA. The cavitary defects were measured on orthogonal views by experienced musculoskeletal radiologists. Radiographically, resorption and trabeculation increased steadily with time. This differential was slightly more noticeable in large defects with a central trabeculation occurring in advance of the peripheral region. The majority of patients progressed to unrestricted activities by 6 weeks and had returned to their usual activities by 12 weeks. No significant difference in graft incorporation rate was noted based on age, size of defect, or use of adjuvant local treatment. The use of a composite graft (ultraporous beta-TCP+BMA) in the treatment of cavitary lesions appears to be safe and effective. PMID- 19292422 TI - Prevalence of knee osteoarthritis and analysis of pain, rigidity, and functional incapacity. AB - Knee osteoarthritis is one of the most prevalent health problems in our society. It accounts for 10% of all primary care visits in general medicine and 30% of outpatient appointments. The objectives of this cross-sectional descriptive study of 100 patients suffering from gonarthritis were to assess pain, functional capacity, and joint damage in patients diagnosed with knee osteoarthritis, as well as the possible repercussions for subsequent surgical treatment. Sociodemographic, clinical, and radiological data were collected, and pain and functional capacity were evaluated by using the Western Ontario and McMaster Universities Osteoarthritis Index. The majority (71) of patients were women, mean age 71 years (SD=7.84), of low educational (66%) and financial (89%) status, with mean disease duration of 11.8 years. Of the total, 87% presented with comorbidity. Radiographs revealed a varus malalignment in 31% of patients and a valgus malalignment in 17%, with bone collapse in 39% of these. The factors that most affect surgery and subsequent rehabilitation are closely linked to social status, the general state of the patient, and the radiological severity of gonarthritis. Most of the patients were obese and suffered from comorbid conditions, and some presented with psychopathology. These factors may influence surgery, and thus improvements in primary care should be made as a way of offering a simpler and more effective treatment for gonarthritis. PMID- 19292423 TI - Factors related to early recovery rate after partial knee meniscectomy. AB - A main role of clinicians treating patients after orthopedic surgery is to inform patients about their anticipated recovery rate and recovery endpoint. In estimating recovery rate, clinicians consider a series of potentially influencing factors, including the type and severity of injury and the characteristics of the patient. Unfortunately, this is done largely without evidence to support factors believed to be important in recovery rate. To our knowledge, no studies exist where factors that might influence recovery rate after arthroscopic partial meniscectomy have been evaluated. Eighty-three patients (11 women and 72 men) were evaluated 4 days after and then again 6 weeks after knee arthroscopic partial meniscectomy surgery. Recovery rate was calculated by dividing the change in a patient's Hughston Clinic knee self-assessment questionnaire during this period by his or her baseline Hughston score and relationships to independent variables (gender, age, body mass index [BMI], injury chronicity, affected meniscus, Hughston Clinic knee self-assessment score at baseline, knee flexion passive range of motion, and knee circumference). These were evaluated using backward stepwise regression analysis. The relationship between recovery rate and the independent variables was statistically significant (P,.05) for the following variables: injury chronicity, gender, and gender/injury type combination. The most unexpected finding in this study was the statistically nonsignificant relationship between recovery rate and the following variables: age, Hughston score, BMI, knee swelling, and knee flexion passive range of motion loss. PMID- 19292424 TI - The biological basis of the bone-muscle inter-relationship in the algorithm of fracture healing. AB - The biological cascade of fracture healing is intimately linked to the muscle envelope. It further depends on the preservation of stable, perpetual axial micromovements. The current study was designed to demonstrate that high molecular weight bioactive substances diffuse from the muscle envelope to initiate osteoinductive activity at experimental fracture sites. Forty-eight rats underwent an experimental fracture of the left tibia and stabilization with an intramedullary 20-gauge needle. The animals were divided into 4 groups (A-D) of 12 rats each according to the post-fracture treatment. In group A (control) no additional treatment was applied following fracture and intramedullary fixation. In groups B, C, and D, a nitrocellulose membrane of various sizes was wrapped around the fracture, separating the periosteum from the muscle envelope. The groups differed by the membrane pore size, allowing passage of the following molecular sizes: 50 kilodaltons (kDa), 12 to 14 kDa, and 3.5 kDa in groups B, C, and D, respectively. Four animals in each group were sacrificed 2, 5, and 10 weeks after the procedure for radiographic and histological evaluation of fracture healing. Radiographic evaluation revealed a decreased rate of bone synthesis that correlated with the nitrocellulose pore size. Morphological and functional analysis of the bone explants indicated poorly healed fractures in groups B, C, and D. Direct contact between fractured bone and its muscle envelope is essential for the biological sequence of new bone formation. The extent of obstruction between the fracture and its muscle envelope correlates with the delay in fracture healing. PMID- 19292428 TI - Morphology and wettability control of silicon cone arrays using colloidal lithography. AB - In this paper we present a simple method to fabricate ordered silicon cone arrays with controllable morphologies on a silicon substrate using reactive ion etching with two-dimensional silica colloidal crystals as masks. The etching process and the morphologies of the obtained structure are quantified. Unlike works reported previously, we show that the surface roughness of the obtained silicon cone arrays can be adjusted by controlling the etching duration, which is proved to be of importance in tailoring the behavior of water droplets when being used as antireflection coatings with superhydrophobicity. Moreover, this strategy is compatible with the methods we have established on controlling the arrangement of colloidal spheres, and thus silicon cone arrays with tunable periodicities, different lattice structures, and various patterns can be prepared. The obtained silicon cone arrays with strips can be used as hydrophobic substrates with anisotropic dewetting just like the leaves of rice. It is found that by adjusting the strip width with and without silicon cones, the water droplets can transform from isotropic dewetting to anisotropic dewetting. PMID- 19292429 TI - Polycondensation and stabilization of chirally ordered molecular organogels derived from alkoxysilyl group- containing L-glutamide lipid. AB - A lipophilic L-glutamide-derived lipid with a triethoxysilyl headgroup (Si-lipid) was newly synthesized as a self-assembling organogelator to stabilize the chirally ordered state of the aggregates. The Si-lipid formed nanofibrous network structures in various organic solvents such as benzene, cyclohexane, and dimethylformamide and entrapped them to form gels. The gels were transformed to sols by heat, and this gel-to-sol transition was thermally reversible. Polycondensation of the triethoxysilyl groups was carried out by acid-catalyzed hydrolysis and condensation in a benzene gel and confirmed by 29Si CP/MAS NMR and FT-IR measurements. After polycondensation, a gel state was maintained, and the thermal and mechanical stabilities of the aggregates increased markedly. Interestingly, polycondensation in chloroform and acetonitrile induced gelation, whereas no gelation was observed before polycondensation. Xerogel, which was prepared by freeze drying organogels, had fibrous network structures similar to those of the original gels. A strong CD signal was observed around the amide bonds in a cyclohexane gel at 20 degrees C, indicating that the gel contained chirally oriented structures based on intermolecular hydrogen bonds. An enhanced CD signal was observed even after polycondensation of the ethoxysilyl group of Si lipid (poly(Si-lipid)) and was maintained at 70 degrees C, which is above the temperature of the gel-to-sol phase transition of the original gel. These results indicate that the formation of siloxane network structure among the fibrous aggregates stabilizes the chiral orientation of lipid aggregates. PMID- 19292430 TI - Selective anion binding in water with use of a zinc(II) dipicolylamino functionalized diketopiperazine scaffold. AB - The design and synthesis of a diketopiperazine based anion receptor bearing two dipicolylamino arms complexed to zinc(II) ions is described. This receptor is readily prepared from the dipeptide precursor by a microwave-assisted intramolecular cyclization reaction. Upon addition of zinc(II), the receptor binds di- and triphosphate ions with high affinity and selectivity in aqueous solution, as determined by using a fluorescent indicator displacement assay. PMID- 19292431 TI - A kinetic model for the enzymatic action of cellulase. AB - We develop a mechanochemical model for the dynamics of cellulase, a two-domain enzyme connected by a peptide linker, as it extracts and hydrolyzes a cellulose polymer from a crystalline substrate. We consider two random walkers, representing the catalytic domain (CD) and the carbohydrate binding module (CBM), whose rates for stepping are biased by the coupling through the linker and the energy required to lift the cellulose polymer from the crystalline surface. Our results show that the linker length and stiffness play a critical role in the cooperative action of the CD and CBM domains and that, for a given linker length, the steady-state hydrolysis shows a maximum at some intermediate linker stiffness. The maximum hydrolysis rate corresponds to a transition of the linker from a compressed to an extended conformation, where the system exhibits maximum fluctuation, as measured by the variance of the separation distance between the two domains and the dispersion around the mean hydrolysis speed. In the range of experimentally known values of the parameters of our model, improving the intrinsic hydrolytic activity of the CD leads to a proportional increase in the overall hydrolysis rate. PMID- 19292433 TI - Time-dependent density functional theory study of the electronic excitation spectra of chlorophyllide a and pheophorbide a in solvents. AB - The electronic excitation spectra of both chlorophyllide a (Chl) and pheophorbide a (Pheo) molecules in solvents have been investigated by using the time-dependent density functional theory (TDDFT) along with the polarizable continuum solvation model (PCM). With increasing Hartree-Fock (HF) exchange percentage in DFT functionals, the predicted HOMO-LUMO gaps increase linearly while the excitation energies increase gradually and even strongly for excited states with partial intramolecular charge-transfer (CT) nature. On the basis of the calculated excitation energies, oscillator strengths and frontier molecular orbital analysis, we provide some new insights into the absorption spectra of Pheo and Chl both in the gas phase and in solutions, especially for the B and higher electronic absorption bands. It is shown that the experimental observed visible Q(y) and Q(x) and ultraviolet B(y) and B(x) bands are all due to singlet (1)(pi,pi*) valence excitations, with the B bands being more strongly red-shifted by solvent effects. Two (1)(pi,pi*) dark states are predicted slightly below (or near) the strong B band for both Chl and Pheo, with one related to the excitation of tetrapyrrole ring and the other related to the excitation of ring I vinyl substituent. The (1)(n,pi*) CT state from the conjugated carbonyl substituent is above B bands and further strongly blue-shifted by solvent effects. The higher eta and N bands are mainly due to (1)(pi,pi*) valence excitations with only partial CT character, which are also red-shifted in solvent. PMID- 19292432 TI - Does water relay play an important role in phosphoryl transfer reactions? Insights from theoretical study of a model reaction in water and tert-butanol. AB - To investigate whether water relay plays an important role in phosphoryl transfer reactions, we have used several theoretical approaches to compare key properties of uridine 3'-m-nitrobenzyl phosphate (UNP) in aqueous and tert-butanol solutions. Previous kinetic experiments found that the isomerization reaction of UNP is abolished in tert-butanol, which was interpreted as the direct evidence that supports the role of water relay in phosphoryl transfer. We have analyzed solute flexibility and solvent structure near the solute using equilibrium molecular dynamics simulations and a combined quantum mechanical/molecular mechanism (QM/MM) potential function for the solute. Snapshots from the simulations are then used in minimum energy path calculations to compare the energetics of direct nucleophilic attack and water-mediated nucleophilic attack pathways. QM/MM simulations are also used to compare the pseudorotation barriers for the pentavalent intermediate formed following the nucleophilic attack, another key step for the isomerization reaction. Combined results from these calculations suggest that water relay does not offer any significant energetic advantage over the direct nucleophilic attack. Unfortunately, the lack of isomerization in tert-butanol solution cannot be straightforwardly explained based on the results we have obtained here and therefore requires additional analysis. This study, nevertheless, has provided new insights into several most commonly discussed possibilities. PMID- 19292434 TI - Preparation and characterization of a pH-responsive nanogel based on a photo cross-linked micelle formed from block copolymers with controlled structure. AB - Poly(ethylene glycol)-b-poly(2-(diethylamino)ethyl methacrylate-co-2 cinnamoyloxyethyl acrylate) (PEG-b-P(DEAEMA/CEA)) was prepared by reversible addition-fragmentation chain transfer (RAFT)-controlled radical polymerization. As solution pH is increased from an acidic pH, the hydrodynamic radius (R(h)) increases abruptly near pH 7, indicative of the micelle formation at pH > 7. The micelle formation at pH > 7 was supported by (1)H NMR and light scattering data. Upon irradiation of light, polymer chains in the core of the polymer micelle are cross-linked as a result of the photodimerization of the cinnamoyl groups, yielding a nanogel. The nanogel was characterized by gel-permeation chromatography (GPC), light scattering, small-angle X-ray scattering (SAXS), transmission electron microscopy (TEM), and fluorescence techniques. The nanogel displayed an ability to solubilize N-phenyl-1-naphthylamine (PNA) and 1 pyrenemethanol (hydrophobic guest molecules) into the hydrophobic core at pH > 7. It was confirmed with PNA that the solubilization of a guest molecule occurred at polymer concentrations (C(p)) lower than the critical micelle concentration (cmc) for PEG-b-P(DEAEMA/CEA) because the nanogel retains its micellar structure at C(p) < cmc. 1-Pyrenemethanol is strongly captured by the nanogel at pH 10, whereas it is easily released from the nanogel when pH is reduced to 3. This indicates that the hydrophobicity of the core of the nanogel can be modulated by a change in the degree of protonation of the DEAEMA units in the core, and thus the capture of a guest molecule and its release can be controlled by a change in solution pH. PMID- 19292435 TI - Spherulitic networks: from structure to rheological property. AB - A finite element method based on ABAQUS is employed to examine the correlation between the microstructure and the elastic response of planar Cayley treelike fiber networks. It is found that the elastic modulus of the fiber network decreases drastically with the fiber length, following the power law. The power law of elastic modulus G' vs the correlation length xi obtained from this simulation has an exponent of -1.71, which is close to the exponent of -1.5 for a single-domain network of agar gels. On the other hand, the experimental results from multidomain networks give rise to a power law index of -0.49. The difference between -1.5 and -0.49 can be attributed to the multidomain structure, which weakens the structure of the overall system and therefore suppresses the increase in G'. In addition, when the aspect ratio of the fiber is smaller than 20, the radius of the fiber cross-section has a great impact on the network elasticity, while, when the aspect ratio is larger than 20, it has almost no effect on the elastic property of the network. The stress distribution in the network is uniform due to the symmetrical network structure. This study provides a general understanding of the correlation between microscopic structure and the macroscopic properties of soft functional materials. PMID- 19292436 TI - Triplet excited states of some thiophene and triazole substituted platinum(II) acetylide chromophores. AB - The photophysical properties of a series of platinum(II) acetylide compounds (trans-Pt(PBu(3))(2)(C[triple bond]C-R)(2)) with the R group consisting of two or three aryl rings (phenyl, phenyl/thiophenyl, phenyl/triazolyl) linked together with ethynyl groups were systematically investigated. Four new structurally similar compounds are reported with: (i) a bithiophene unit in the ligands, (ii) methyl or (iii) methoxy substituents on the aryl ring ligands that promote a more twisted conformation along the long axis of the molecule, and (iv) with two different alkynylaryl ligands giving rise to an asymmetric substitution with respect to the photoactive metal ion center. The spectroscopic studies include optical absorption, spectrally and time-resolved luminescence, as well as transient absorption spectra. The ground-state UV absorption between 300 and 420 nm gave rise to fluorescence with quantum efficiencies in the range of 0.1-1% and efficient intersystem crossing to triplet states. Phosphorescence decay times were in the order of 10-500 micros in oxygen-evacuated samples. The triplet states also lead to strong broadband triplet-triplet absorption between 400 and 800 nm. The complex with asymmetric substitution was found to populate two triplet states of different structure and energy. PMID- 19292437 TI - Biosynthesis of aromatic polyketides in bacteria. AB - Natural products, produced chiefly by microorganisms and plants, can be large and structurally complex molecules. These molecules are manufactured by cellular assembly lines, in which enzymes construct the molecules in a stepwise fashion. The means by which enzymes interact and work together in a modular fashion to create diverse structural features has been an active area of research; the work has provided insight into the fine details of biosynthesis. A number of polycyclic aromatic natural products--including several noteworthy anticancer, antibacterial, antifungal, antiviral, antiparasitic, and other medicinally significant substances--are synthesized by polyketide synthases (PKSs) in soil borne bacteria called actinomycetes. Concerted biosynthetic, enzymological, and structural biological investigations into these modular enzyme systems have yielded interesting mechanistic insights. A core module called the minimal PKS is responsible for synthesizing a highly reactive, protein-bound poly-beta ketothioester chain. In the absence of other enzymes, the minimal PKS also catalyzes chain initiation and release, yielding an assortment of polycyclic aromatic compounds. In the presence of an initiation PKS module, polyketide backbones bearing additional alkyl, alkenyl, or aryl primer units are synthesized, whereas a range of auxiliary PKS enzymes and tailoring enzymes convert the product of the minimal PKS into the final natural product. In this Account, we summarize the knowledge that has been gained regarding this family of PKSs through recent investigations into the biosynthetic pathways of two natural products, actinorhodin and R1128 (A-D). We also discuss the practical relevance of these fundamental insights for the engineered biosynthesis of new polycyclic aromatic compounds. With a deeper understanding of the biosynthetic process in hand, we can assert control at various stages of molecular construction and thus introduce unnatural functional groups in the process. The metabolic engineer affords a number of new avenues for creating novel molecular structures that will likely have properties akin to their fully natural cousins. PMID- 19292439 TI - Preparation, characterization, and oral delivery of insulin loaded carboxylated chitosan grafted poly(methyl methacrylate) nanoparticles. AB - To improve the efficiency of insulin via oral administration, pH-sensitive carboxylated chitosan grafted poly(methyl methacrylate) nanoparticles (CCGN) were prepared. CCGN were characterized by (1)H NMR, dynamic light scattering, zeta potential, and transmission electron microscopy, and the hypoglycemic effect of insulin loaded CCGN via the oral route was evaluated in normal and diabetic rats. CCGN exhibited a homogeneous morphology and a spherical shape with core-shell structure. They were aggregated in simulated gastric fluid while separated in simulated intestinal fluid. Insulin was mainly located in the shell of the CCGN via hydrogen bonding, electrostatic interaction, and Van der Waals force. Insulin release from the CCGN exhibited a pH-sensitive property in that it had a slow release rate at pH 2.0 and a fast release rate at pH 6.8 and 7.4. The pharmacological bioavailability after oral administration of insulin loaded CCGN at a dose of 25 IU/kg was found to be 9.7%. Besides, CCGN showed desirable tissue and blood compatibility. Therefore, the CCGN would be a promising delivery carrier for protein drugs via the oral route. PMID- 19292441 TI - One-pot, high-yield synthesis of 5-fold twinned Pd nanowires and nanorods. AB - In the presence of poly(vinylpyrrolidone) and I(-), uniform Pd nanowires and nanorods have been successfully prepared. As-prepared one-dimensional Pd nanostructures have a 5-fold twinned structure. The twinned Pd nanowires can be further transformed into nanowires/nanorods with a smaller aspect ratio and finally to multiply twinned particles upon increasing the reaction time. With structural anisotropy, the one-dimensional Pd nanostructures exhibit different chemical reactivities on their tips and sides. PMID- 19292443 TI - Dietary administration of high doses of pterostilbene and quercetin to mice is not toxic. AB - The aim of this study is to evaluate possible harmful effects of high doses of t pterostilbene (t-PTER) and quercetin (QUER) in Swiss mice. Mice were fed during 28 days at doses of 0, 30, 300, and 3000 mg/kg body weight/day of t-PTER, QUER, or a mixture of both, t-PTER + QUER, which are equivalent to 5, 50, and 500 times, respectively, the estimated mean human intake of these polyphenols (25 mg/day). Daily oral administration of QUER, t-PTER, or a mixture of both of them did not cause mortality during the experimental period. There were no differences in food and water consumption on sex. No significant body weight gain in the male or female groups was observed. Red blood cell number and the hematocrit increased after polyphenols administration compared to control groups. Biochemical parameters were not affected. Histopathological examination revealed no alterations in clinical signs or organ weight at any dose. PMID- 19292442 TI - Slow hydrogen atom transfer reactions of oxo- and hydroxo-vanadium compounds: the importance of intrinsic barriers. AB - Reactions are described that interconvert vanadium(IV) oxo-hydroxo complexes [V(IV)O(OH)(R(2)bpy)(2)]BF(4) (1a-c) and vanadium(V) dioxo complexes [V(V)O(2)(R(2)bpy)(2)]BF(4) (2a-c) [R(2)bpy = 4,4'-di-tert-butyl-2,2'-bipyridine ((t)Bu(2)bpy), a; 4,4'-dimethyl-2,2'-bipyridine (Me(2)bpy), b; 2,2'-bipyridine (bpy), c]. These are rare examples of pairs of isolated, sterically unencumbered, first-row metal-oxo/hydroxo complexes that differ by a hydrogen atom (H(+) + e( )). The V(IV)-(t)Bu(2)bpy derivative 1a has a useful (1)H NMR spectrum, despite being paramagnetic. Complex 2a abstracts H(*) from organic substrates with weak O H and C-H bonds, converting 2,6-(t)Bu(2)-4-MeO-C(6)H(2)OH (ArOH) and 2,2,6,6 tetramethyl-N-hydroxypiperidine (TEMPOH) to their corresponding radicals ArO(*) and TEMPO, hydroquinone to benzoquinone, and dihydroanthracene to anthracene. The equilibrium constant for 2a + ArOH <==> 1a + ArO(*) is (4 +/- 2) x 10(-3), implying that the VO-H bond dissociation free energy (BDFE) is 70.6 +/- 1.2 kcal mol(-1). Consistent with this value, 1a is oxidized by 2,4,6 (t)Bu(3)C(6)H(2)O(*). All of these reactions are surprisingly slow, typically occurring over hours at ambient temperatures. The net hydrogen-atom pseudo-self exchange 1a + 2b <==> 2a + 1b, using the (t)Bu- and Me-bpy substituents as labels, also occurs slowly, with k(se) = 1.3 x 10(-2) M(-1) s(-1) at 298 K, DeltaH(double dagger) = 15 +/- 2 kcal mol(-1), and DeltaS(double dagger) = 16 +/- 5 cal mol(-1) K. Using this k(se) and the BDFE, the vanadium reactions are shown to follow the Marcus cross relation moderately well, with calculated rate constants within 10(2) of the observed values. The vanadium self-exchange reaction is ca. 10(6) slower than that for the related Ru(IV)O(py)(bpy)(2)(2+)/Ru(III)OH(py)(bpy)(2)(2+) self-exchange. The origin of this dramatic difference has been probed with DFT calculations on the self exchange reactions of 1c + 2c and on monocationic ruthenium complexes with pyrrolate or fluoride in place of the py ligands. The calculations reproduce the difference in barrier heights and show that transfer of a hydrogen atom involves more structural reorganization for vanadium than the Ru analogues. The vanadium complexes have larger changes in the metal-oxo and metal-hydroxo bond lengths, which is traced to the difference in d-orbital occupancy in the two systems. This study thus highlights the importance of intrinsic barriers in the transfer of a hydrogen atom, in addition to the thermochemical (bond strength) factors that have been previously emphasized. PMID- 19292444 TI - Monitoring secondary structural changes in salted and smoked salmon muscle myofiber proteins by FT-IR microspectroscopy. AB - Fourier transform infrared (FT-IR) microspectroscopy and light microscopy were used to study changes in the myofibrillar proteins and microstructure in salmon muscle due to dry salting and smoking. Light microscopy showed that the myofibers of the smoked samples were more shrunken and their shape more irregular and edged than for the nonsmoked samples. FT-IR microspectroscopy showed that salting time mostly contributed in the amide I region, revealing that secondary structural changes of proteins were primarily affected by salting. The main variation in the amide II region was caused by smoking. As it is known that smoke components can react with amino acid side chains and that the contribution of the side chain in the amide II region is larger than that in amide I, it is concluded that the observed differences are due to interactions between carbonyl compounds of smoke and amino acid side chains. PMID- 19292445 TI - Charge redistribution in oxidized and semiquinone E. coli DNA photolyase upon photoexcitation: stark spectroscopy reveals a rationale for the position of Trp382. AB - The electronic structure of the two lowest excited electronic states of FAD and FADH(*) in folate-depleted E. coli DNA photolyase (PL(OX) and PL(SQ), respectively) was measured using absorption Stark spectroscopy. The experimental analysis was supported by TDDFT calculations of both the charge redistribution and the difference dipole moments for the transitions of both oxidation states using lumiflavin as a model. The difference dipole moments and polarizabilities for PL(OX) are similar to those obtained in our previous work for flavins in simple solvents and in an FMN-containing flavoprotein. No such comparison can be made for PL(SQ), as we believe this to be the first experimental report of the direction and magnitude of excited-state charge redistribution in any flavosemiquinone. The picture that emerges from these studies is discussed in the context of electron transfer in photolyase, particularly for the semiquinone photoreduction process, which involves nearby tryptophan residues as electron donors. The direction of charge displacement derived from an analysis of the Stark spectra rationalizes the positioning of the critical Trp382 residue relative to the flavin for efficient vectorial electron transfer leading to photoreduction. The ramifications of vectorial charge redistribution are discussed in the context of the wider class of flavoprotein blue light photoreceptors. PMID- 19292446 TI - Hydration of protonated aromatic amino acids: phenylalanine, tryptophan, and tyrosine. AB - The first steps of hydration of the protonated aromatic amino acids phenylalanine, tryptophan, and tyrosine were studied experimentally employing a mass spectrometer equipped with a drift cell to examine the sequential addition of individual water molecules in equilibrium experiments and theoretically by a combination of molecular mechanics and electronic structure calculations (B3LYP/6 311++G**) on the three amino acid systems including up to five water molecules. It is found that both the ammonium and carboxyl groups offer good water binding sites with binding energies of the order of 13 kcal/mol for the first water molecule. Subsequent water molecules bind less strongly, in the range of 7-11 kcal/mol for the second through fifth water molecules. The ammonium group is able to host up to three water molecules and the carboxyl group one water molecule before additional water molecules bind either to the amino acid side chain as in tyrosine or to already-bound water in a second solvation shell around the ammonium group. Reasons for the surprisingly high water affinity of the neutral carboxyl group, comparable to that of the charge-carrying ammonium group, are found to be high intrinsic hydrophilicity, favorable charge-dipole alignment, and -for the case of multiply hydrated species--favorable dipole-dipole interaction among water molecules and the lack of alternative fully exposed hydration sites. PMID- 19292447 TI - Transition states of Plasmodium falciparum and human orotate phosphoribosyltransferases. AB - Orotate phosphoribosyltransferases (OPRT) catalyze the formation of orotidine 5' monophosphate (OMP) from alpha-D-phosphoribosylpyrophosphate (PRPP) and orotate, an essential step in the de novo biosynthesis of pyrimidines. Pyrimidine de novo biosynthesis is required in Plasmodium falciparum , and thus OPRT of the parasite (PfOPRT) is a target for antimalarial drugs. De novo biosynthesis of pyrimidines is also a feature of rapidly proliferating cancer cells. Human OPRT (HsOPRT) is therefore a target for neoplastic and autoimmune diseases. One approach to the inhibition of OPRTs is through analogues that mimic the transition states of PfOPRT and HsOPRT. The transition state structures of these OPRTs were analyzed by kinetic isotope effects (KIEs), substrate specificity, and computational chemistry. With phosphonoacetic acid (PA), an analogue of pyrophosphate, the intrinsic KIEs of [1'-(14)C], [1, 3-(15)N(2)], [3-(15)N], [1'-(3)H], [2'-(3)H], [4'-(3)H], and [5'-(3)H(2)] are 1.034, 1.028, 0.997, 1.261, 1.116, 0.974, and 1.013 for PfOPRT and 1.035, 1.025, 0.993, 1.199, 1.129, 0.962, and 1.019 for HsOPRT, respectively. Transition state structures of PfOPRT and HsOPRT were determined computationally by matching the calculated and intrinsic KIEs. The enzymes form late associative D(N)*A(N)(double dagger) transition states with complete orotate loss and partially associative nucleophile. The C1'-O(PA) distances are approximately 2.1 A at these transition states. The modest [1' (14)C] KIEs and large [1'-(3)H] KIEs are characteristic of D(N)*A(N)(double dagger) transition states. The large [2'-(3)H] KIEs indicate a ribosyl 2'-C-endo conformation at the transition states. p-Nitrophenyl beta-D-ribose 5'-phosphate is a poor substrate of PfOPRT and HsOPRT but is a nanomolar inhibitor, supporting a reaction coordinate with strong leaving group activation. PMID- 19292448 TI - Zeptomol detection through controlled ultrasensitive surface-enhanced Raman scattering. AB - SERS permits identifying the nature of molecules in extremely low concentrations, but it is hindered by poor enhancement or low reproducibility. We demonstrate controllable approximately 10(10) signal amplification reaching the zeptomol detection limit for a nonresonant molecule by sandwiching the analyte between the tips of star-shaped gold nanoparticles and a planar gold surface using a simple synthetic procedure. This unprecedented control over light-intensity amplification opens a new avenue toward high-yield, fully reproducible, SERS based, zeptomol detection and holds promise for nonlinear optics applications at the single-particle level. PMID- 19292449 TI - Direct detection of molecular biorecognition by dipole sensing mechanism. AB - This work investigates the feasibility of transducing molecular-recognition events into a measurable potentiometric signal. It is shown for the first time that biorecognition of acetylcholine (ACh) can be translated to conformational changes in the enzyme, acetylcholine-esterase (AChE), which in turn induces a measurable change in surface potential. Our results show that a highly sensitive detector for ACh can be obtained by the dilute assembly of AChE on a floating gate derived field effect transistor (FG-FET). A wide concentration range response is observed for ACh (10(-2)-10(-9)M) and for the inhibitor carbamylcholine CCh (10(-6)-10(-11)M). These enhanced sensitivities are modeled theoretically and explained by the combined response of the device to local pH changes and molecular dipole variations due to the enzyme-substrate recognition event. PMID- 19292450 TI - Total synthesis of vinblastine, vincristine, related natural products, and key structural analogues. AB - Full details of the development of a direct coupling of catharanthine with vindoline to provide vinblastine are described along with key mechanistic and labeling studies. Following an Fe(III)-promoted coupling reaction initiated by generation of a presumed catharanthine radical cation that undergoes a subsequent oxidative fragmentation and diastereoselective coupling with vindoline, addition of the resulting reaction mixture to an Fe(III)-NaBH(4)/air solution leads to oxidation of the C15'-C20' double bond and reduction of the intermediate iminium ion directly providing vinblastine (40-43%) and leurosidine (20-23%), its naturally occurring C20' alcohol isomer. The yield of coupled products, which exclusively possess the natural C16' stereochemistry, approaches or exceeds 80% and the combined yield of the isomeric C20' alcohols is >60%. Preliminary studies of Fe(III)-NaBH(4)/air oxidation reaction illustrate a generalizable trisubstituted olefin scope, identify alternatives to O(2) trap at the oxidized carbon, provide a unique entry into C20' functionalized vinblastines, and afford initial insights into the observed C20' diastereoselectivity. The first disclosure of the use of exo-catharanthine proceeding through Delta(19',20') anhydrovinblastine in such coupling reactions is also detailed with identical stereochemical consequences. Incorporating either a catharanthine N-methyl group or a vindoline N-formyl group precludes Fe(III)-promoted coupling, whereas the removal of the potentially key C16 methoxy group of vindoline does not adversely impact the coupling efficiency. Extension of these studies provided a total synthesis of vincristine (2) via N-desmethylvinblastine (36, also a natural product), 16-desmethoxyvinblastine (44) and 4-desacetoxy-16-desmethoxyvinblastine (47) both of which we can now suggest are likely natural products produced by C. roseus, desacetylvinblastine (62) and 4-desacetoxyvinblastine (59), as well as a series of key analogues bearing systematic modifications in the vindoline subunit. Their biological evaluation provided additional insights into the key functionality within the vindoline subunit contributing to the activity and sets the foundation on which further, more deep-seated changes in the structures of 1 and 2 will be explored in future studies. PMID- 19292451 TI - One-pot synthesis of interpenetrating inorganic/organic networks of CuO/resorcinol-formaldehyde aerogels: nanostructured energetic materials. AB - For many applications ranging from catalysis to sensors to energetic materials, it is desirable to produce intimate mixtures of nanoparticles. For instance, to improve the reaction rates of energetic materials, the oxidizing agent and the fuel need to be mixed as intimately as possible, ideally at the nanoscopic level. In this context, the acidity of a hydrated CuCl(2) solution reacting toward a network of CuO nanoparticles (a good oxidant) is used to induce one-pot cogelation of a nanostructured network of a resorcinol-formaldehyde resin (RF, the fuel). The resulting wet gels are dried to aerogels, and upon pyrolysis under Ar, the interpenetrating CuO/RF network undergoes a smelting reaction toward metallic Cu. Upon ignition in the open air, pure RF aerogels do not burn, while CuO/RF composites, even with substoichiometric CuO, sustain combustion, burning completely leaving only a solid residue of CuO whose role then has been that of a redox mediator through the smelting reaction. PMID- 19292452 TI - Lacticin 481 synthetase as a general serine/threonine kinase. AB - Methods that introduce posttranslational modifications in a general, mild, and non-sequence-specific manner using biologically produced peptides have great utility for investigation of the functions of these modifications. In this study, the substrate promiscuity of a lantibiotic synthetase was exploited for the preparation of phosphopeptides, glycopeptides, and peptides containing analogs of methylated or acetylated lysine residues. Peptides attached to the C-terminus of the leader peptide of the lacticin 481 precursor peptide were phosphorylated on serine residues in a wide variety of sequence contexts by the R399M and T405A mutants of lacticin 481 synthetase (LctM). Serine residues located as many as 30 amino acids C-terminal to the leader peptide were phosphorylated. Wild-type LctM was shown to dehydrate these peptides to generate dehydroalanine-containing products that can be conveniently modified with external nucleophiles including thiosaccharides, 2-(dimethylamino)ethanethiol, and N-acetyl cysteamine, resulting in mimics of O-linked glycopeptides and acetylated and methylated lysines. PMID- 19292453 TI - A novel mechanism is involved in cationic lipid-mediated functional siRNA delivery. AB - A key challenge for therapeutic application of RNA interference is to efficiently deliver synthetic small interfering RNAs (siRNAs) into target cells that will lead to the knockdown of the target transcript (functional siRNA delivery). To facilitate rational development of nonviral carriers, we have investigated by imaging, pharmacological and genetic approaches the mechanisms by which a cationic lipid carrier mediates siRNA delivery into mammalian cells. We show that approximately 95% of siRNA lipoplexes enter the cells through endocytosis and persist in endolysosomes for a prolonged period of time. However, inhibition of clathrin-, caveolin-, or lipid-raft-mediated endocytosis or macropinocytosis fails to inhibit the knockdown of the target transcript. In contrast, depletion of cholesterol from the plasma membrane has little effect on the cellular uptake of siRNA lipoplexes, but it abolishes the target transcript knockdown. Furthermore, functional siRNA delivery occurs within a few hours and is gradually inhibited by lowering temperatures. These results demonstrate that although endocytosis is responsible for the majority of cellular uptake of siRNA lipoplexes, a minor pathway, probably mediated by fusion between siRNA lipoplexes and the plasma membrane, is responsible for the functional siRNA delivery. Our findings suggest possible directions for improving functional siRNA delivery by cationic lipids. PMID- 19292454 TI - Identification and characterization of wolframin, the product of the wolfram syndrome gene (WFS1), as a novel calmodulin-binding protein. AB - To search for calmodulin (CaM) targets, we performed affinity chromatography purification of a rat brain extract using CaM fused with GST as the affinity ligand. Proteomic analysis was then carried out to identify CaM-binding proteins. In addition to identifying 36 known CaM-binding proteins, including CaM kinases, calcineurin, nNOS, the IP(3) receptor, and Ca(2+)-ATPase, we identified an ER transmembrane protein, wolframin [the product of the Wolfram syndrome gene (WFS1)] as interacting. A CaM overlay and an immunoprecipitation assay revealed that wolframin is capable of binding the Ca(2+)/CaM complex in vitro and in transfected cells. Surface plasmon resonance analysis and zero-length cross linking showed that the N-terminal cytoplasmic domain (residues 2-285) of wolframin binds to an equimolar unit of CaM in a Ca(2+)-dependent manner with a K(D) for CaM of 0.15 muM. Various truncation and deletion mutants showed that the Ca(2+)/CaM binding region in wolframin is located from Glu90 to Trp186. Furthermore, we demonstrated that three mutations (Ala127Thr, Ala134Thr, and Arg178Pro) associated with Wolfram syndrome completely abolished CaM binding of wolframin. This observation may indicate that CaM binding is important for wolframin function and that impairment of this interaction by mutation contributes to the pathology seen in Wolfram syndrome. PMID- 19292455 TI - Oxidative disassembly of the [2Fe-2S] cluster of human Grx2 and redox regulation in the mitochondria. AB - Mitochondrial Grx2 is a new member of the thioredoxin superfamily that has been found to bind a [2Fe-2S] cluster in a novel coordination motif at the interface of a homodimer, where cluster binding occurs via a catalytic cysteine residue and a molecule of GSH (per monomer). The (Grx2)(2)-[2Fe-2S] dimer is thought to undergo cluster destruction and monomerization in a redox-induced pathway of activation. In this report, we make use of protein film voltammetry (PFV) as a method to probe the stability of the Grx2-[2Fe-2S] cluster, using oxidative poises of varying potential and duration to probe the thermodynamic and kinetic stability of the cluster's electrochemical response. We find that the cluster signal is stable at positive potentials up to 0.5 V but that cluster destruction occurs readily when oxidative pulses in excess of this value are applied. PMID- 19292456 TI - Thermodynamics of multivalent carbohydrate-lectin cross-linking interactions: importance of entropy in the bind and jump mechanism. AB - The high affinity (K(d) = 0.2 nM) of the soybean agglutinin (SBA), a tetrameric GalNAc specific lectin, for a modified form of porcine submaxillary mucin, a linear glycoprotein, with a molecular mass of approximately 10(6) Da and approximately 2300 GalNAcalpha1-O-Ser/Thr residues (Tn-PSM) has been ascribed to an internal diffusion mechanism that involves binding and jumping of the lectin from GalNAc to GalNAc residue of the mucin [Dam, T. K., et al. (2007) J. Biol. Chem. 282, 28256-28263]. Hill plot analysis of the raw ITC data shows increasing negative cooperativity, which correlates with an increasing number of lectin mucin cross-linking interactions and decreasing favorable binding entropies. However, the affinity of bound SBA for other Tn-PSM molecules during cross linking is much higher than that of free SBA for GalNAcalpha1-O-Ser, a monovalent analogue. The high affinity of bound SBA for GalNAc residues on other Tn-PSM molecules appears to be due to the favorable entropy of binding associated with the internal diffusion mechanism. Furthermore, the increasing negative cooperativity of SBA binding to Tn-PSM correlates with a decreasing level of internal diffusion of the lectin on the mucin as cross-linking occurs. These findings indicate the importance of the internal diffusion mechanism in generating large, favorable entropies of binding that drive lectin-mucin cross linking interactions. The results are important for understanding the energetics of lectin-mucin cross-linking interactions that are associated with biological signaling on the surface of cells and the role of the internal diffusion mechanism in ligand-biopolymer interactions in general. PMID- 19292457 TI - Cholesterol exposure at the membrane surface is necessary and sufficient to trigger perfringolysin O binding. AB - Perfringolysin O (PFO) is the prototype for the cholesterol-dependent cytolysins, a family of bacterial pore-forming toxins that act on eukaryotic membranes. The pore-forming mechanism of PFO exhibits an absolute requirement for membrane cholesterol, but the complex interplay between the structural arrangement of the PFO C-terminal domain and the distribution of cholesterol in the target membrane is poorly understood. Herein we show that PFO binding to the bilayer and the initiation of the sequence of events that culminate in the formation of a transmembrane pore depend on the availability of free cholesterol at the membrane surface, while changes in the acyl chain packing of the phospholipids and cholesterol in the membrane core, or the presence or absence of detergent resistant domains do not correlate with PFO binding. Moreover, PFO association with the membrane was inhibited by the addition of sphingomyelin, a typical component of membrane rafts in cell membranes. Finally, addition of molecules that do not interact with PFO, but intercalate into the membrane and displace cholesterol from its association with phospholipids (e.g., epicholesterol), reduced the amount of cholesterol required to trigger PFO binding. Taken together, our studies reveal that PFO binding to membranes is triggered when the concentration of cholesterol exceeds the association capacity of the phospholipids, and this cholesterol excess is then free to associate with the toxin. PMID- 19292458 TI - Enantioselective phytoeffects of chiral pesticides. AB - Chirality exists extensively in nature. Synthetic chiral plant growth regulators and other pesticides usually behave enantioselectively in phyto-biochemical processes. Chiral plant growth regulators regulate the physiological processes of plants enantioselectively, and chiral pesticides cause enantioselective toxicities or ecotoxicities to plants. On the other hand, these chiral agrochemicals can be absorbed and enantioselectively metabolized by plants. This review summarizes the enantioselective effects of chiral plant growth regulators on plants and the phytotoxic and biotransformation effects of chiral herbicides and several persistent organic pollutants (POPs) on plants. Together, this information on the interactions between chiral agrochemicals and plants might shed light on studies on the chemical and biological behaviors of chiral chemicals, and direct research into the selection of plants, which can potentially decontaminate the environment. PMID- 19292459 TI - Development of an analytical scheme for the determination of pyrethroid pesticides in composite diet samples. AB - Analysis of an individual's total daily food intake may be used to determine aggregate dietary ingestion of given compounds. However, the resulting composite sample represents a complex mixture, and measurement of such can often prove to be difficult. In this work, an analytical scheme was developed for the determination of 12 select pyrethroid pesticides in dietary samples. In the first phase of the study, several cleanup steps were investigated for their effectiveness in removing interferences in samples with a range of fat content (1 10%). Food samples were homogenized in the laboratory, and preparatory techniques were evaluated through recoveries from fortified samples. The selected final procedure consisted of a lyophilization step prior to sample extraction. A sequential 2-fold cleanup procedure of the extract included diatomaceous earth for removal of lipid components followed with a combination of deactivated alumina and C(18) for the simultaneous removal of polar and nonpolar interferences. Recoveries from fortified composite diet samples (10 microg kg( 1)) ranged from 50.2 to 147%. In the second phase of this work, three instrumental techniques [gas chromatography-microelectron capture detection (GC microECD), GC-quadrupole mass spectrometry (GC-quadrupole-MS), and GC-ion trap MS/MS] were compared for greatest sensitivity. GC-quadrupole-MS operated in selective ion monitoring (SIM) mode proved to be most sensitive, yielding method detection limits of approximately 1 microg kg(-1). The developed extraction/instrumental scheme was applied to samples collected in an exposure measurement field study. The samples were fortified and analyte recoveries were acceptable (75.9-125%); however, compounds coextracted from the food matrix prevented quantitation of four of the pyrethroid analytes in two of the samples considered. PMID- 19292460 TI - A thorough study on the use of quantitative 1H NMR in Rioja red wine fermentation processes. AB - In this study, we focused our attention on monitoring the levels of important metabolites of wine during the alcoholic and malolactic fermentation processes by quantitative nuclear magnetic resonance (qNMR). Therefore, using (1)H NMR, the method allows the simultaneous quantification of ethanol, acetic, malic, lactic, and succinic acids, and the amino acids proline and alanine, besides the ratio proline/arginine through fermentation of must of grapes corresponding to the Tempranillo variety. Each (1)H NMR spectrum gives direct and visual information concerning these metabolites, and the effectiveness of each process was assessed and compared by carrying out analyses using infrared spectroscopy to ethanol and acetic acid. The quantitative data were explained with the aid of chemometric algorithms. PMID- 19292461 TI - Development and validation of a duplex real-time PCR method to simultaneously detect potentially allergenic sesame and hazelnut in food. AB - The paper describes the development and validation of a duplex real-time PCR method allowing the simultaneous detection of traces of potentially allergenic sesame and hazelnut in food. For the detection of sesame and hazelnut, the genes coding for two major allergenic proteins, Ses i 1 and Cor a 1, were selected. The duplex real-time PCR assay did not show any cross-reactivity with 25 common food ingredients from sesame and/or hazelnut containing foods. Analysis of serially diluted sesame/hazelnut DNA resulted in good linearity up to a dilution of 1:10000 (corresponding to 10 pg microL(-1) or 50 pg). Sesame and hazelnut could be detected in blank whole meal cookies which had been spiked with 0.005% sesame and 0.005% hazelnut. The applicability of the real-time PCR assay for determining sesame and hazelnut in different food matrices was investigated by analyzing 30 commercial foodstuffs comprising salty snacks, cookies, chocolates, creams, mueslis and muesli bars. PMID- 19292462 TI - Quantification of ferritin from staple food crops. AB - Ferritin-iron has been shown to be as bioavailable as ferrous sulfate in humans. Thus, biofortification to breed crops with high ferritin content is a promising strategy to alleviate the global iron deficiency problem. Although ferritin is present in all food crops, its concentration varies between species and varieties. Therefore, a successful ferritin biofortification strategy requires a method to rapidly measure ferritin concentrations in food crops. The objective of this study was to develop a simple and reliable ELISA using an anti-ferritin polyclonal antibody to detect ferritin in various crops. Crude seed extracts were found to have 10.2 +/- 1.0, 4.38 +/- 0.9, 1.2 +/- 0.3, 0.38 +/- 0.1, and 0.04 +/- 0.01 microg of ferritin/g of dry seed in red beans, white beans, wheat, maize, and brown rice, respectively. Although the measured absolute concentrations of ferritin values were low, the presented method is applicable for rapid screening for the relative ferritin concentrations of large numbers of seeds to identify and breed ferritin-rich crops. PMID- 19292463 TI - Protein expression in orthocortical and paracortical cells of merino wool fibers. AB - Crimp and bulk, important wool fiber properties, are thought to be related to differences in the protein composition of the orthocortex and paracortex. Fiber morphological studies have demonstrated that the paracortex has a higher proportion of matrix and cysteine than the orthocortex. While there is some evidence for the differential expression of genes between these cell types in the follicle, this has not been demonstrated satisfactorily in the mature fiber. Using proteolytic digestion of wool fibers, followed by ultrasonic disruption to obtain relatively pure fractions of both cell types, the KAP3 high sulfur protein family was found to be present in higher concentrations in the paracortex. This significant finding provides an explanation for the higher cysteine content reported in the paracortex. This represents an advance in our understanding of protein expression variation in the orthocortex and paracortex, and how this relates to key physical and mechanical properties of wool fibers. PMID- 19292464 TI - Antiproliferation effect and mechanism of prostate cancer cell lines as affected by isoflavones from soybean cake. AB - The objectives of this study were to determine the antiproliferation effect of prostate cancer cell lines LNCaP and PC-3 as affected by 4 isoflavone fractions prepared from soybean cake and isoflavone standards genistein and daidzein. With the MTT test, most treatments were effective in inhibiting prostate cancer cell growth at a low dose of 5 and 10 mug/mL. In cell cycle analysis, the fractions of aglycon, a mixture of acetylglucoside and aglycon, as well as genistein and a combination of genistein and daidzein standards exhibited a high G2/M ratio for LNCaP, as did the acetylglucoside, genistein and a combination of genistein and daidzein standards for PC-3. Results of Western blotting revealed an increase in p53 protein expression of LNCaP following treatments of the aglycon fraction, genistein and a combination of genistein and daidzein standards. However, all the treatments did not affect Bcl-2 protein expression significantly in both LNCaP and PC-3 cells. A decline in cyclin B1 expression of LNCaP was observed for all the treatments, with the mixture of acetylglucoside and aglycon possessing the most pronounced effect. But for PC-3, a decrease in cyclin B1 expression was shown for all the isoflavones, with the exception of malonylglucoside, glucoside and acetylglucoside fractions. The outcome of this study may provide a basis for possible production of functional food in the future with soybean cake as raw material. PMID- 19292465 TI - Characterization of total, free and esterified phytosterols in tetraploid and hexaploid wheats. AB - Dietary plant sterols have received increasing attention in recent years due to their favorable health benefits. The present research focused on quantification of phytosterols as free, esterified and total forms in different tetraploid (5 cultivars of Triticum durum Desf., 9 cultivars of Triticum dicoccon Schrank) and hexaploid (5 cultivars of T. aestivum L., 12 cultivars of Triticum spelta L.) wheats. Tetraploid wheats showed the highest content of total sterol (79.4 and 79.5 mg of sterols /100 g dry weight for T. durum and T. dicoccon, respectively). Hexaploid cultivars were the best source of esterified sterols (40.7% and 37.3% of total sterols for Triticum aestivum and T. spelta, respectively). Significant amounts of free sterols (65.5% and 60.7% of total sterols for T. durum and T. dicoccon, respectively) were found in the tetraploid cultivars. The most abundant phytosterol in all wheat samples was sitosterol accounting for 45.1-59.1, 46.6 57.4 and 38.6-59.5% of total, free and esterified sterol fraction, respectively. These results demonstrate that although the sterol profile present in tetraploid and hexaploid wheat species is the same, differences in their relative amounts and distribution allow statistical differentiation between hexaploids and tetraploids, and between soft and durum wheats. PMID- 19292466 TI - Ovicidal and adulticidal activities of Origanum majorana essential oil constituents against insecticide-susceptible and pyrethroid/malathion-resistant Pediculus humanus capitis (Anoplura: Pediculidae). AB - The toxicity of essential oil constituents from marjoram, Origanum majorana, to eggs and adult females of the susceptible KR-HL and dual malathion- and permethrin-resistant BR-HL strains of human head louse, Pediculus humanus capitis, was examined using contact + fumigant mortality bioassay. Results were compared with those following treatment with two pyrethroid pediculicides, d phenothrin or pyrethrum. As judged by the lethal time to 50% mortality (LT(50)) values at the exposure rate of 0.25 mg/cm(2), 1,8-cineole (14.1 min) was the most toxic compound, followed by linalool (15.4 min) to KR-HL females. These compounds were faster acting than either d-phenothrin (24.1 min) or pyrethrum (33.4 min). Based on the lethal concentration causing 50% mortality (LC(50)) values, (-) camphor (0.022 mg/cm(2)) was the most toxic compound, followed by linalool (0.035 mg/cm(2)), (-)-terpinen-4-ol (0.040 mg/cm(2)), alpha-terpineol (0.045 mg/cm(2)), and 1,8-cineole (0.068 mg/cm(2)) against KR-HL females. These monoterpenoids were less toxic than either d-phenothrin (LC(50), 0.0015 mg/cm(2)) or pyrethrum (0.0013 mg/cm(2)). However, the toxicities of these monoterpenoids were almost identical against females from either of the two strains, even though the BR-HL females exhibited high levels of resistance to d-phenothrin [resistance ratio (RR), 667] and pyrethrum (RR, 754). After a 24 h exposure to linalool, BR-HL egg hatch was inhibited 100 and 84% at 0.25 or 0.125 mg/cm(2), respectively, while ( )-terpinen-4-ol caused 94 and 69% inhibition of egg hatch at 0.25 and 0.125 mg/cm(2). alpha-Terpineol caused 88 and 76% inhibition of egg hatch at 0.5 and 0.25 mg/cm(2), respectively. Thus, certain monoterpenoids from O. majorana essential oil, particularly linalool, (-)-terpinen-4-ol and alpha-terpineol, merit further study as potential pediculicides and ovicides for the control of insecticide-resistant P. h. capitis populations as fumigants with contact action. PMID- 19292467 TI - Residue-free wines: fate of some quinone outside inhibitor (QoI) fungicides in the winemaking process. AB - The fate of three fungicide residues (fenamidone, pyraclostrobin, and trifloxystrobin) from vine to wine was studied to evaluate the decay ratio and the influence of the technological process. The aim of this work was to identify pesticides that can degrade rapidly or be eliminated together with byproduct (lees and cake) of the winemaking process to obtain wine free of residues. The disappearance rate on grapes was calculated as pseudo-first-order kinetics, and the half-life (t(1/2)) was in the range from 5.4 +/- 1.9 to 12.2 +/- 1.2 days. The mechanism of dissipation of the three quinone outside inhibitor (QoI) fungicides was studied using different model systems. It was observed that the main mechanism responsible for disappearance was photodegradation. For active ingredients (ai) the half-lives of fenamidone, pyraclostrobin, and trifloxystrobin were 10.2 +/- 0.8, 20.1 +/- 0.1, and 8.6 +/- 1.0 h, respectively, whereas for formulation higher half-lives were observed when epicuticular waxes were present (from 13.8 +/- 0.2 to 26.6 +/- 0.1 h). After winemaking, fenamidone, pyraclostrobin, and trifloxystrobin residues were not detected in the wine, but they were present in the cake and lees. This was due to the adsorption of pesticide residues to the solid parts, which are always eliminated at the end of the alcoholic fermentation. The data obtained in these experiments suggest that these three active ingredients could be used in a planning process to obtain residue-free wines. PMID- 19292468 TI - Effect of boiling on the content of ascorbigen, indole-3-carbinol, indole-3 acetonitrile, and 3,3'-diindolylmethane in fermented cabbage. AB - The aim of the study was to investigate the effect of the boiling process on the content of ascorbigen, indole-3-carbinol, indole-3-acetonitrile, and 3,3' diindolylmethane in fermented cabbage. The cabbage was boiled for 5 to 60 min. Boiling resulted in a decrease of the total content of the compounds analysed. The changes were mainly caused by leaching of ascorbigen predominating in cabbage into cooking water and by its thermal hydrolysis. Ascorbigen losses resulting from thermal hydrolysis accounted for 30% after 10 min of boiling and for 90% after 60 min of boiling. One of the ascorbigen breakdown products was indole 3 carbinol; the decrease in ascorbigen content was accompanied by a drastic increase in the content of 3,3'-diindolylmethane, a condensation product of indole-3-carbinol. After 40 and 50 min of boiling, the total content of 3,3' diindolylmethane in cabbage and cooking water was approximately 0.2 micromol/100 g and was 6-fold higher than that in uncooked cabbage. 3,3'-Diindolylmethane synthesis proceeded within the plant tissue. After 10 min of boiling, the content of free indole-3-carbinol and indole-3-acetonitrile stabilized at the level of about 80% as compared to the uncooked cabbage. PMID- 19292469 TI - Persistence of two neem formulations on peach leaves and fruit: effect of the distribution. AB - Persistence of azadirachtins (A+B) and of the other limonoids (nimbin, salannin, deacetylnimbin, and deacetylsalannin) on peach leaves and fruits was studied using a commercial formulation (form. C) compared with an experimental formulation (form. E) prepared with coformulations allowed in organic culture. Field experiments were carried out using three concentrations: 1x, 5x, and 10x the dose recommended by the manufacturer. The EU maximum residue level (MRL) in fruits and vegetables for azadirachtin A is 1 mg/kg with a preharvest interval (PHI) of 3 days. At the recommended dose, azadirachtin A residue on fruits was not detectable (LOQ < 0.8 microg/kg). After field treatment at the 5x concentration, azadirachtoids were found with 22% in the epicuticular waxes and the remaining 78% on the fruit surface. No residues were found in the fruit pulp. The experimental formulation (E) produced lower residues on leaves and fruit compared with the commercial formulation (C), although formulation E showed greater stability. This is probably due to the amount of the active ingredients that diffuse into the epicuticular wax layer thus enhancing photostability of azadirachtoids. PMID- 19292470 TI - First identification of O,S-diethyl Thiocarbonate in Indian Cress absolute and odor evaluation of its synthesized homologues by GC-sniffing. AB - Indian cress (Tropaeolum majus L.) absolute was studied by GC-olfactometry (VIDEO Sniff method) in order to identify odor-active aroma compounds. Because of its fruity-sulfury odor note, a compound that has never been identified in plant extracts before stood out: O,S-diethyl thiocarbonate, present at 0.1% (percentage of the total GC/FID area) in the extract. GCxGC-TOFMS allowed for a clean mass spectrum to be obtained, and isolation by preparative GC followed by NMR studies allowed its identification. Here, we report on the first detection of O,S-diethyl thiocarbonate in Indian cress absolute by GC-olfactometry/VIDEO-Sniff and on its isolation and identification. The synthesis and odor evaluation of its homologues are presented. PMID- 19292471 TI - Biodegradable zein-based films: influence of gamma-irradiation on structural and functional properties. AB - Zein, a predominant corn protein, is an alcohol-soluble protein extracted from corn and is characterized by unique film-forming properties. The characteristic brittleness of zein diminishes its usefulness as a structural material. The objective of this work was to study the effect of gamma-irradiation on improving the performance of zein films in packaging applications. This goal has been achieved by irradiating zein film-forming solutions with various doses of gamma rays, namely, 10, 20, 30, and 40 kGy at dose rate of 10.5 kGy/h, using a Co(60) gamma-radiation source. The impact of radiation process on the structural properties has been explored through far-UV CD spectral analysis and scanning electron microscopy. Additionally, viscosity changes that reflect the effect of radiation treatment on degradation and/or cross-linking of protein chains have been measured. However, improvements in the performance of zein films as packaging materials that can be accomplished by radiation treatment have been investigated via monitoring of the color, surface density, roughness, mechanical properties (tensile strength and elongation percentage), water uptake, and water barrier properties. The results indicated that gamma-irradiation treatment of the film-forming solution can be used to improve the water barrier properties, as well as color and appearance of the zein films. Moreover, a sterilization effect is considered to be an additional advantage for applying gamma- irradiation. PMID- 19292472 TI - Adenosine, an identified active component from the Driselase-treated fraction of rice bran, is effective at improving metabolic syndrome in stroke-prone spontaneously hypertensive rats. AB - In the present study, we isolated and identified an active component from the Driselase-treated fraction and investigated its effect by acute and chronic oral administration on hypertension, lipid, and glucose metabolism in stroke-prone spontaneously hypertensive rats. The active component was identified as adenosine and improves hypertension after single oral administration. Rats who were 10 weeks old were divided into control and adenosine groups and were administered water or water with adenosine (10 mg/L), respectively, for 3 weeks. Hypertension and plasma lipid, nitric oxide, insulin, leptin, adiponectin levels, and glucose metabolism were significantly improved in the adenosine group. The mRNA expression levels of genes involved in lipid and glucose metabolism were altered in the adenosine group. Single oral administration of adenosine (10 mg/kg body weight) improved hypertension and plasma triglyceride, glucose, and nitric oxide levels 2 h after administration. In conclusion, oral acute and chronic administration of adenosine are beneficial and improve the metabolic syndrome related disease parameters. PMID- 19292473 TI - Protective effect of red beetroot against carbon tetrachloride- and N nitrosodiethylamine-induced oxidative stress in rats. AB - The aim of the study was to investigate the potential protective effect of beetroot juice in a model of oxidative stress induced by N-nitrosodiethylamine (NDEA) and carbon tetrachloride (CCl(4)). Male Wistar rats were treated with beetroot juice per os, 8 mL/kg/day for 28 days, and a single i.p. dose of the xenobiotics: 150 mg/kg NDEA or 2 mL/kg CCl(4). Simultaneously, two groups of rats not pretreated with juice were given only each of the xenobiotics. The level of microsomal lipid peroxidation in the liver, expressed as TBARS concentration, was increased several fold in rats administered only NDEA or CCl(4). TBARS were decreased by 38% only in rats pretreated with beetroot juice before the administration of CCl(4). In animals pretreated with juice and receiving NDEA, a further increase in TBARS occurred. All of the investigated antioxidant enzymes were inhibited by the administration of either toxicant alone by 26%-77% as compared to controls. Pretreatment with juice caused a partial recovery in the activity of glutathione peroxidase and glutathione reductase, by 35% and 66%, respectively. Superoxide dismutase activity was increased about 3-fold in animals pretreated with juice. Both xenobiotics caused a rise in plasma protein carbonyls, which were reduced by 30% in rats pretreated with juice and then injected with NDEA. Similarly, DNA damage in blood leukocytes caused by either toxicant was slightly diminished, by 20%, in the rats treated with juice before NDEA administration. It could be concluded that pretreatment with beetroot juice can counteract, to some extent, xenobiotic-induced oxidative stress in rats. PMID- 19292474 TI - Chronic ingestion of apple pectin can enhance the absorption of quercetin. AB - The effect of apple pectin (AP) on quercetin and rutin bioavailability was investigated. Rats received a basal (control) or AP-containing diet for 6 weeks followed by orally administered quercetin or rutin. With quercetin administration, the maximum concentration and area under the curve of concentration-time from 0 to 24 h for the sum of quercetin metabolites in the plasma were significantly higher in AP-fed rats than in the control group. However, AP did not significantly affect rutin bioavailability. The crypt depth of the jejunum and ileum and the villus thickness of the ileum of AP-fed rats were significantly greater than those of control rats. These results demonstrate that chronic AP ingestion enhances intestinal absorption of quercetin. This increase in quercetin absorption might be attributed to alteration of the absorptive capacity of the small intestine through AP-induced improvement of its morphological and physiological properties. PMID- 19292475 TI - Inhibitory effects of grape seed procyanidins on foam cell formation in vitro. AB - Human and animal studies have demonstrated that procyanidin-rich diets reduce the risk of cardiovascular diseases and atherosclerosis. Some beneficial effects have been attributed to the well-known antioxidant activity of procyanidins. This study investigated another potential corrective role of procyanidins in cholesterol flux and inflammation in macrophage-derived foam cells. RAW 264.7 macrophages were cultured with moderately oxidized LDL (oxLDL), minimally oxidized LDL (moxLDL), or LPS (0.5 microg/mL) and oxLDL (LPS + oxLDL) to induce foam cells. Then, cells were treated with procyanidins derived from grape seed (PE, 45 microg/mL) for the last 12 h of incubation with the different lipoproteins (25 microg/mL). After lipid extraction, it was determined that total and esterified cholesterol and triglyceride accumulations in foam cells were increased by lipoprotein treatment but reduced by PE incubation. To asses the effect of PE on gene expression, the relative mRNA levels of CD36, ABCA1, iNOS, COX-2, and IkappaBalpha were determined by RT-PCR. It was shown that PE reduced the oxLDL scavenger receptor expression (CD36) and enhanced ATP-binding cassette A1 (ABCA1) expression, a key regulator of macrophage cholesterol efflux. PE also down-regulated inflammatory-related genes such as inducible nitric oxide synthase (iNOS) and kappa beta inhibitor-alpha (IkappaBalpha) without modifying COX-2 expression. In conclusion, evidence is provided that procyanidins may attenuate the development of foam cell formation by reducing cholesterol accumulation and modulating the expression of key genes in cholesterol flux and inflammation. PMID- 19292476 TI - Editorial: "Conjugates of oligonucleotides and modified oligonucleotides": a memorable review by John Goodchild. PMID- 19292477 TI - A two-step binding model proposed for the electrostatic interactions of ricin a chain with ribosomes. AB - Ricin is a ribosome inactivating protein that catalytically removes a universally conserved adenine from the alpha-sarcin/ricin loop (SRL) of the 28S rRNA. We recently showed that ricin A chain (RTA) interacts with the P1 and P2 proteins of the ribosomal stalk to depurinate the SRL in yeast. Here we examined the interaction of RTA with wild-type and mutant yeast ribosomes deleted in the stalk proteins by surface plasmon resonance. The interaction between RTA and wild-type ribosomes did not follow a single-step binding model but was best characterized by two distinct types of interactions. The AB1 interaction had very fast association and dissociation rates, was saturable, and required an intact stalk, while the AB2 interaction had slower association and dissociation rates, was not saturable, and did not require the stalk. RTA interacted with the mutant ribosomes by a single type of interaction, which was similar to the AB2 interaction with the wild-type ribosomes. Both interactions were dominated by electrostatic interactions, and the AB1 interaction was stronger than the AB2 interaction. On the basis of these results, we propose a two-step interaction model. The slow and ribosomal stalk nonspecific AB2 interactions concentrate the RTA molecules on the surface of the ribosome. The AB2 interactions facilitate the diffusion of RTA toward the stalk and promote the faster, more specific AB1 interactions with the ribosomal stalk. The electrostatic AB1 and AB2 interactions work together allowing RTA to depurinate the SRL at a much higher rate on the intact ribosomes than on the naked 28S rRNA. PMID- 19292478 TI - Nucleic acids: expanding the structural and functional horizons. PMID- 19292479 TI - High-throughput thermal scanning: a general, rapid dye-binding thermal shift screen for protein engineering. AB - The low stability of natural proteins often limits their use in therapeutic, industrial, and research applications. The scale and throughput of methods such as circular dichroism, fluorescence spectroscopy, and calorimetry severely limit the number of variants that can be examined. Here we demonstrate a high throughput thermal scanning (HTTS) method for determining the approximate stabilities of protein variants at high throughput and low cost. The method is based on binding to a hydrophobic dye akin to ANS, which fluoresces upon binding to molten globules and thermal denaturation intermediates. No inherent properties of the protein, such as enzymatic activity or presence of an intrinsic fluorophore, are required. Very small sample sizes are analyzed using a real-time PCR machine, enabling the use of high-throughput purification. We show that the apparent T(M) values obtained from HTTS are approximately linearly related to those from CD thermal denaturation for a series of four-helix bundle hydrophobic core variants. We demonstrate similar results for a small set of TIM barrel variants. This inexpensive, general, and scaleable approach enables the search for conservative, stable mutants of biotechnologically important proteins and provides a method for statistical correlation of sequence-stability relationships. PMID- 19292480 TI - Monodispersed polymeric nanocapsules: spontaneous evolution and morphology transition from reducible hetero-PEG PICmicelles by controlled degradation. AB - In this communication, a novel "self-templating" strategy was used to prepare uniform and biocompatible nanocapsules by the addition of a reduction agent (i.e., DTT) into a solution of highly monodispersed PICmicelles bearing a heterodetachable PEG corona. PEG chains were released from PICmicelle shells following disulfide reduction which leads a spontaneous and drastic morphology evolution from micelles to vesicles induced by the decrease of the PEG weight fraction. Formation of uniform nanocapsules with controllable capsule size was achieved by careful control of the micelle composition and molecular weight of homo-P[Asp(DET)]. PMID- 19292481 TI - Stabilization of RNA oligomers through reverse micelle encapsulation. AB - The cellular milieu is rich in diversity of both simple and complex molecules and is also quite crowded. By contrast, typical sample concentrations employed for in vitro investigation of biophysics and structural biology make use of purified macromolecules in simple buffer systems at concentrations that range from micromolar to millimolar. Although this formulation has proven to be compatible with a wide range of biological and structural studies, it is quite different from the relatively crowded conditions typically found within cells. The importance of these crowding effects for proteins has been recognized for some time, but the equivalent analysis is underexplored in nucleic acids. Encapsulation with surfactant-based reverse micelles has emerged as an effective biophysical tool, allowing study of the influence of ionic strength, pH, hydration, and crowding on biologically active macromolecules over a wide range of conditions. We have encapsulated an oligonucleotide model of TAR RNA from HIV and the 5' stem loop oligonucleotide of the U4 snRNA. Observation of imino (1)H resonances is an established method for evaluating the stability of nucleic acid oligonucleotides, implying the presence of stacked, hydrogen bonded base pairs. Inspection of (1)H NMR spectra of the RNA molecules reveals that the intensity of several of the imino resonances increases upon encapsulation. Additional resonances not observed in spectra of the oligonucleotides free in solution support the suggestion that the molecules have gained stability. These results indicate that RNA oligonucleotides may acquire significant stability in the presence of cellular levels of crowding. PMID- 19292482 TI - Toward an accurate determination of free energy landscapes in solution states of proteins. AB - The dynamics of proteins plays a central role in their activity, including enzymatic catalysis and allosteric communication. Many advances have been made in recent years in the characterization of the equilibrium fluctuations of proteins through experimental and computational methods. We present evidence that the use of molecular dynamics simulations with ensemble-averaged structural restraints derived from nuclear magnetic resonance spectroscopy enables the determination of ensembles of structures representing the equilibrium populations of conformations explored during the thermal fluctuations of proteins. We obtained these results by using residual dipolar couplings to characterize the dynamics of ubiquitin and to derive its free-energy landscape under native conditions. PMID- 19292483 TI - Timing of the Delta(10,12)-Delta(11,13) double bond migration during ansamitocin biosynthesis in Actinosynnema pretiosum. AB - The timing of introduction of the unusually placed Delta(11,13) diene system in ansamitocin (AP) biosynthesis was probed by synthesizing optically active potential tri- and tetraketide intermediates as their SNAC thioesters. An AP nonproducing mutant Actinosynnema pretiosum was complemented by the R enantiomer of the triketide and by the tetraketide with rearranged double bonds, but not by the tetraketide carrying the double bonds in conjugation to the thioester function. The results show that the double bonds are installed in their final positions during processing of the nascent polyketide on module 3 of the asm PKS and that KS4 of the PKS acts as a gatekeeper which accepts only a tetraketide with shifted double bonds as substrate for further processing. PMID- 19292484 TI - Isolation of notoamide E, a key precursor in the biosynthesis of prenylated indole alkaloids in a marine-derived fungus, Aspergillus sp. AB - Notoamide E was identified to be a short-lived precursor in the biosynthesis of prenylated indole alkaloids in the mussel-derived Aspergillus sp. In addition, the feeding experiment of the (13)C-labeled notoamide E afforded structurally novel metabolites. PMID- 19292485 TI - Palladium-catalyzed diacetoxylation of alkenes with molecular oxygen as sole oxidant. AB - A new palladium-catalyzed diacetoxylation of alkenes using oxygen as the sole oxidant to afford diacetates was developed. High levels of diastereoselectivity in diacetoxylation of 1,2-disubstituted alkenes was obtained. PMID- 19292486 TI - Binding and enantiomeric selectivity of threonyl-tRNA synthetase. AB - A combination of MD simulations and free energy calculations have been used to propose a new model for the binding of amino acids to threonyl-tRNA-synthetase which not only yields a stable binding mode for l-Ser but also can explain the mechanism by which the editing domains of aminoacyl-tRNA-synthetases are enantiomeric selective preferentially binding d-amino acids. PMID- 19292487 TI - Post-synthesis alkoxide formation within metal-organic framework materials: a strategy for incorporating highly coordinatively unsaturated metal ions. AB - A new noncatenated metal-organic framework containing pendant alcohol functionalities was synthesized. The alcohols were then post-synthetically converted to either lithium or magnesium alkoxides, with the incorporated metals anchored far from nodes or carboxylate functionalities. The metal alkoxide sites can be obtained stoichiometrically while maintaining the permanent porosity and large surface area of the parent hydroxylated material. The incorporated metal ions are found to induce an unusual pattern of binding energetics for H(2): isosteric heats of adsorption increase, rather than decrease, with increasing H(2) loading. Additionally, at 1 atm and 77 K, uptake (at least with low Li(+) loading) is increased by two hydrogen molecules per Li(+). PMID- 19292488 TI - Control of pore size and functionality in isoreticular zeolitic imidazolate frameworks and their carbon dioxide selective capture properties. AB - Five new crystalline zeolitic imidazolate frameworks (ZIFs), ZIF-78 to -82, were prepared from zinc(II) nitrate and mixtures of 2-nitroimidazole and five different functionalized imidazoles and were found to have the GME topology. These structures, along with three previously reported GME ZIFs, constitute a series of highly porous materials with Brunauer-Emmet-Teller surface areas ranging from 620 to 1730 m(2)/g. The pore diameters and apertures vary incrementally from 7.1 to 15.9 A and 3.8 to 13.1 A, respectively, and the functionalities decorating the pores vary from polar cyano- and nitro- groups to nonpolar alkyl groups. The variability expressed in these materials makes them highly attractive for study as gas-separation media. Selectivity values calculated for separation of CO(2) and CH(4) predict that the ZIFs with polar functionality, ZIF-78 (10.6:1) and -82 (9.6:1), retain CO(2) gas to a greater degree than the other members of the GME series and BPL-activated carbon. These predictions are borne out in dynamic breakthrough studies, which confirm the increased capacity of ZIF-78 and -82 and demonstrate the promise of this class of materials. PMID- 19292489 TI - Both intra- and interstrand charge-transfer excited states in aqueous B-DNA are present at energies comparable to, or just above, the (1)pipi* excitonic bright states. AB - Vertical electronic excitations in model systems representing single- and double stranded B-DNA are characterized using electronic structure theory, including both time-dependent density functional theory (TD-DFT) and correlated wave function techniques. Previous TD-DFT predictions of charge-transfer (CT) states well below the optically bright (1)pipi* states are shown to be artifacts of the improper long-range behavior of standard density-functional exchange approximations, which we rectify here using a long-range correction (LRC) procedure. For nucleobase dimers (hydrogen-bonded or pi-stacked), TD-LRC-DFT affords vertical excitation energies in reasonable agreement with the wave function methods, not only for the (1)npi* and (1)pipi* states but also for the CT states, and qualitatively reproduces well-known base-stacking effects on the absorption spectrum of DNA. The emergence of (1)pipi* Frenkel exciton states, localized on a single strand, is clearly evident, and these states (rather than low-energy CT states) are primarily responsible for the fact that DNA's absorption spectrum exhibits a red tail that is absent in monomer absorption spectra. For B-DNA in aqueous solution, the low-energy tail of the CT band (representing both intra- and interstrand CT states) appears at energies comparable to those of the optically bright (1)pipi* exciton states. In systems with more than one base pair, we also observe the emergence of delocalized, interstrand CT excitations, whose excitation energies may be significantly lower than the lowest CT excitation in a single base pair. Together, these observations suggest that a single Watson-Crick base pair is an inadequate model of the photophysics of B-DNA. PMID- 19292490 TI - Synthesis and polymerase chain reaction amplification of DNA strands containing an unnatural triazole linkage. AB - DNA strands containing an unnatural T-triazole-T linkage have been synthesized by click DNA ligation between oligonucleotides with 3'-AZT and 5'-propargylamido dT and amplified efficiently by polymerase chain reaction (PCR) using several different polymerases. DNA sequencing of PCR amplicons and clones in two different sequence contexts revealed the presence of a single thymidine at the ligation site. The remarkable ability of thermostable polymerases to reproducibly copy DNA templates containing such an unnatural backbone opens up intriguing possibilities in gene synthesis, genetic analysis, biology, and nanotechnology. PMID- 19292491 TI - Acyl radical reactions in fullerene chemistry: direct acylation of [60]fullerene through an efficient decatungstate-photomediated approach. AB - A versatile and highly efficient photochemical methodology for the direct acylation of C(60) has been developed. This approach utilizes a wide variety of acyl radicals derived from aldehydes through a hydrogen atom abstraction process mediated by tetrabutylammonium decatungstate [(n-Bu(4)N)(4)W(10)O(32)]. The single addition reaction of these acyl radicals to [60]fullerene proceeded selectively to afford a novel class of previously unexplored fullerene-based materials. Product analysis of this reaction showed that decarbonylation and acylation pathways compete when a tertiary or phenylacetyl aldehyde is the starting material. However, a decrease of the reaction temperature was found to be effective in overcoming the decarbonylation encountered in certain acyl radical additions to C(60); the carbonyl radical addition precedes decarbonylation even in the cases where the decarbonylation rate constant exceeds 10(6) s(-1) (i.e., phenylacetaldehyde). The regiochemistry of the t-butyl radical addition was also found to be thermally controlled. The present methodology is directly applicable even in the cases of the cyclopropyl-substituted aldehydes, where rapid rearrangement of the cyclopropyl acyl radical intermediate can potentially occur. A mechanistic approach for this new reactivity of C(60) has been also provided, based mainly on intra- and intermolecular deuterium isotope effect studies. PMID- 19292492 TI - Structures of thermal, mass-selected water clusters probed with hydrophobic ion tags and infrared photodissociation spectroscopy. AB - The structures of tetramethylammonium (TMA(+)) and phenyltrimethylammonium (PTMA(+)) with 1-8 water molecules attached were investigated using infrared photodissociation spectroscopy (IRPD), blackbody infrared radiative dissociation, and theory to elucidate the effects of ion-water interactions and water-water hydrogen bonding in clusters containing these hydrophobic ions. Several pieces of evidence suggest that these ions have only a small effect on water structure. The binding energy of a water molecule to TMA(+) is 44 kJ/mol, a value that is significantly lower than that reported for other cations and close to the condensation energy of bulk water. The OH asymmetric stretch of the water molecule in TMA(+).(H(2)O)(1) at 3718 cm(-1) is less red-shifted from that of an isolated water molecule (3756 cm(-1)) than for those of other cations, and water water hydrogen bonding is extensive for all clusters with two or more water molecules. These results indicate that charge transfer from water to these hydrophobic ions is much less than that for other cations. There are two predominant structures for these ions with three water molecules: one in which water molecules form a chain and one with a cyclic structure analogous to that of neutral water trimer. With four water molecules attached, water forms a cyclic structure similar to that of neutral water tetramer. The spectra of the larger clusters follow trends reported for neutral water molecule clusters, which indicates that these hydrophobic ions have only a minimal effect on the structure of water in clusters with four or more water molecules. These results suggest that these hydrophobic ions can be used as "tags" that could make possible acquisition of IRPD spectra of even larger clusters, such as clathrates, for which precise mass selection of neutral clusters is not possible by other methods. PMID- 19292493 TI - Mechanistic insights on the hydrodesulfurization of biphenyl-2-thiol with nickel compounds. AB - The reactivity of the nickel(I) dimer [(dippe)Ni(mu-H)](2) (1) with biphenyl-2 thiol was explored with the aim of clarifying the key step of sulfur extrusion during the hydrodesulfurization process using dibenzothiophene (DBT). These reactions were monitored by variable temperature NMR experiments which allowed the complete characterization and isolation of [(dippe)(2)Ni(2)(mu-H)(mu-S-2 biphenyl)] (3). The latter compound evolves to the terminal nickel-hydride [(dippe)Ni (eta(1)-C-2-biphenyl)(H)] (4) and transient [(dippe)NiS] (5), to ultimately yield [(dippe)(2)Ni(2)(mu-S)] (2) and biphenyl as the resulting HDS products. The reactivity of 1 and biphenyl-2-thiol was examined using different ratios of reactants, which allowed preparation of [(dippe)Ni(eta(1)-S-biphenyl-2 thiolate)(2)] (6) when using an excess of this substrate. The reactivity of 6 with 1 was addressed, yielding compound 2 and an equivalent amount of biphenyl. PMID- 19292494 TI - Dinuclear Zn(II) complex promotes cleavage and isomerization of 2-hydroxypropyl alkyl phosphates by a common cyclic phosphate intermediate. AB - The kinetics and cleavage products of 2-hydroxypropyl p-nitrophenyl phosphate were determined in methanol containing the di-Zn(II) complex of bis-1,3-N1,N1' (1,5,9-triazacyclododecyl)propane (4). Time-dependent 1H NMR spectra of the reaction mixture at sspH 9.8 +/- 0.1 show that the catalytic reaction proceeds via a cyclic phosphate (4-methylethylene phosphate, 2) that is subsequently cleaved into a kinetic mixture of two isomeric products, 2-hydroxypropyl methyl phosphate (3) and 1-hydroxypropan-2-yl methyl phosphate (3a), in a 29/71 ratio. In the presence of 4, the kinetic mixture of 3/3a is transformed into a thermodynamic mixture of 72/28 3/3a. The time-dependent 1H NMR spectra of 4 and a 22/78 mixture of 3/3a in CD3OH show that the formation of the thermodynamic mixture occurs on the same time scale as replacement of the P-OCH3 group of the 3/3a starting materials with OCD3. Detailed kinetic studies indicate that the dominant process for loss of the OCH3 group and equilibration of 3/3a is via a 4 catalyzed process where each of the isomers cyclizes to methylethylene phosphate (2), which subsequently reforms the 3/3a thermodynamic mixture. The kcatmax for 4 catalyzed cyclization of 3 and three other 2-hydroxypropyl O-alkyl phosphates (alkyl = CF3CH2- (6a), CH2FCH2- (6b), and CH3CH2- (6c)) has been determined, and the Bronsted plot comprising the log kcatmax vs leaving group sspKa that includes several previously studied 2-hydroxypropyl aryl phosphates is linear, following the expression log kcatmax = (-0.85 +/- 0.02) sspKa + (12.8 +/- 0.4). The betalg value of -0.85 suggests that the catalyzed cleavage of the P-OAr/OR bond has progressed to about 45% in the transition state. The combined results are analyzed in terms of two possible processes involving either a concerted reaction leading to the cyclic phosphate 2 from which the thermodynamic mixture of 3/3a is formed or a stepwise one involving a transient phosphorane whose predominant fate is to eliminate methoxide and proceed to 2 rather than partitioning between 3, 3a, and 2. PMID- 19292498 TI - Charting the chemical space of target sites: insights into the binding modes of amine and amidine groups. AB - Nowadays there is growing awareness that the translation of the increasing number of lead compounds into clinical candidates is still a slow and often inefficient process. In order to facilitate the lead optimization procedure, due consideration must be given to the use of the right bioisosteric replacements. Very recently, we reported that exploring a chemical space of binding sites is a more effective strategy for studying the bioisosteric relationships existing among functional groups. As a continuation of our work in this field, we report herein the construction of a chemical space covered by binding sites of small molecules containing diverse amine and amidine groups. The analysis of the differences in some properties of the binding sites of these functional groups allow for gaining insights into the binding modes of positively charged groups. In addition, this study pinpoints that different types of interactions and bioisosteric relationships exist among primary, secondary, tertiary, quaternary amine, and amidine moieties. PMID- 19292499 TI - Direct correlation of structures and nanomechanical properties of multicomponent lipid bilayers. AB - Exploring the fine structures and physicochemical properties of physiologically relevant membranes is crucial to understanding biological membrane functions including membrane mechanical stability. We report a direct correlation of the self-organized structures exhibited in phase-segregated supported lipid bilayers consisting of dioleoylphosphatidylcholine/egg sphingomyelin/cholesterol (DEC) in the absence and presence of ceramide (DEC-Ceramide) with their nanomechanical properties using AFM imaging and high-resolution force mapping. Direct incorporation of ceramide into phase-segregated supported lipid bilayers formed ceramide-enriched domains, where the height topography was found to be imaging setpoint dependent. In contrast, liquid ordered domains in both DEC and DEC Ceramide presented similar heights regardless of AFM imaging settings. Owing to its capability for simultaneous determination of the topology and interaction forces, AFM-based force mapping was used in our study to directly correlate the structures and mechanical responses of different coexisting phases. The intrinsic breakthrough forces, regarded as fingerprints of bilayer stability, along with elastic moduli, adhesion forces, and indentation of the different phases in the bilayers were systematically determined on the nanometer scale, and the results were presented as two-dimensional visual maps using a self-developed code for force curves batch analysis. The mechanical stability and compactness were increased in both liquid ordered domains and fluid disordered phases of DEC Ceramide, attributed to the influence of ceramide in the organization of the bilayer, as well as to the displacement of cholesterol as a result of the generation of ceramide-enriched domains. The use of AFM force mapping in studying phase segregation of multicomponent lipid membrane systems is a valuable complement to other biophysical techniques such as imaging and spectroscopy, as it provides unprecedented insight into lipid membrane mechanical properties and functions. PMID- 19292500 TI - Metabonomics approach to understanding acute and chronic stress in rat models. AB - The effects of acute and chronic stress on the production of systemic metabolites were investigated in male Sprague-Dawley (SD) rats. Metabolites excreted in urine were analyzed using GC/MS in conjunction with multivariate and univariate statistical techniques. SD rats were subjected to two kinds of acute stress and chronic unpredictable mild stress, respectively. Metabolic analysis demonstrated that urinary expression of a number of metabolites including glutamate, glutamine, homovanillate, proline, succinate, citrate, and tyrosine altered in the acute stress model in the same way as in the chronic model, while pimelate and hippurate changed in the opposite trend. The results suggested that the stress induced metabolic perturbations were reversible and nonspecific. Metabolic response to chronic combined stress revealed biochemical clues to depression-like symptoms validated by behavior and physiologic results. This study provides a noninvasive and dynamic analytical strategy for the characterization of endogenous metabolic perturbations induced by external stress. PMID- 19292501 TI - Kinetic aspects of emulsion stabilization by surfactants: a microfluidic analysis. AB - In classical emulsification processes, surfactants play two roles: first, they reduce the interfacial tension, facilitating droplet deformation and rupture, and second, they reduce droplet coalescence. Here, we use a microfluidic emulsification system to completely uncouple these two processes, allowing stabilization against coalescence to be studied quantitatively and independently of droplet formation. We demonstrate that, in addition to the classical effect of stabilization by an increase of surfactant concentration, the dynamics of adsorption of surfactant at the water-oil interface is a key element for droplet stabilization. Microfluidic emulsification devices can therefore be tailored to improve emulsification while decreasing the concentration of surfactant by increasing the time before the droplets first come into contact. PMID- 19292502 TI - Comprehensive clinico-glycomic study of 16 colorectal cancer specimens: elucidation of aberrant glycosylation and its mechanistic causes in colorectal cancer cells. AB - The structures of neutral and acidic glycosphingolipids from both normal colorectal epithelial cells and colorectal cancer cells, which were highly purified with the epithelial cell marker CD326, have been analyzed. The analysis was performed on samples from 16 patients. The carbohydrate moieties from glycosphingolipids were released by endoglycoceramidase II, labeled by pyridylamination, and identified using two-dimensional mapping and mass spectrometry. The structures from normal colorectal epithelial cells are characterized by dominant expression of neutral type-1 chain oligosaccharides. Three specific alterations were observed in malignant transformation; increased ratios of type-2 oligosaccharides, increased alpha2-3 and/or alpha2-6 sialylation and increased alpha1-2 fucosylation. Although the degree of alteration varies case to case, we found that two characteristic alterations tend to be associated with clinical features. One is a shift from type-1 dominant normal colorectal epithelial cells to type-2 dominant colorectal cancer cells. This shift was found in 5 patients having hepatic metastasis. The other is specific elevation of alpha2-3 sialylation observed in 2 cases exhibiting high serum levels of CA19-9. Examination of the activities of the related glycosyltransferases revealed that while some alterations could be accounted for by changes in the activities of related glycosyltransferases others could not. Although the number of cases analyzed is small, these findings provide valuable information which will help in the elucidation of the mechanism of synthesis of aberrant glycosylation and its involvement in cancer malignancy. PMID- 19292503 TI - Hybrid thermoreversible gels from covalent polymers and organogels. AB - This paper reports on experiments intended for investigating the feasibility of preparing hybrid thermoreversible gels from covalent polymers and noncovalent self-assembling pi-conjugated molecules. The formation and the degree of dispersion of these hybrid gels have been studied with polystyrenes of various tacticities and oligo(p-phenylenevinylene) molecules (OPV) in different nonpolar organic solvents. Detailed investigations of the systems have been carried out by DSC, SAXS, and AFM. It is shown that no liquid-liquid phase separation is involved, indicating that the systems are highly compatible, and that the growth of one type of gel does not interfere with the other. These studies reveal that the resultant hybrid gels are composed of the intermingled fibrillar architectures of both gels. PMID- 19292504 TI - Polymer-assisted crystallization and optical properties of uniform microrods of organic dye Sudan II. AB - The self-assembly of uniform single-crystalline microrods of organic dye Sudan II (1-(2,4- dimethylphenyl-azo)-2-naphthol) in solution was achieved by reprecipitation with the assistance of amphiphilic block copolymer Pluronic F127. It was found that the formation of F127 micelles was favorable for the production of uniform Sudan II microrods whereas individual F127 molecules favored the formation of Sudan II fibers. The optical absorption and fluorescence emission properties of the uniform microrods were characterized, and the time-dependent spectra of the precipitating solution for microrod formation were measured to monitor the self-assembly process of Sudan II molecules. The Sudan II microrods exhibited intense fluorescence emission, indicating an unusual aggregation induced emission (AIE) phenomenon for Sudan II. The fluorescent microscopy investigation suggested that the obtained Sudan II microrods exhibited remarkable optical waveguide behavior, which is expected to serve as single rod active optical waveguides that can be used as building blocks for miniaturized photonic devices. Furthermore, novel Sudan II microplates, which exhibited interesting 2D optical waveguide behavior, were readily obtained by reprecipitation of Sudan II from cationic surfactant cetyltrimethylammonium bromide (CTAB) solution. PMID- 19292505 TI - Control entropy: a complexity measure for nonstationary signals. AB - We propose an entropy statistic designed to assess the behavior of slowly varying parameters of real systems. Based on correlation entropy, the method uses symbol dynamics and analysis of increments to achieve sufficient recurrence in a short time series to enable entropy measurements on small data sets. We analyze entropy along a moving window of a time series, the entropy statistic tracking the behavior of slow variables of the data series. We employ the technique against several physiological time series to illustrate its utility in characterizing the constraints on a physiological time series. We propose that changes in the entropy of measured physiological signal (e.g. power output) during dynamic exercise will indicate changes in underlying constraint of the system of interest. This is compelling because CE may serve as a non-invasive, objective means of determining physiological stress under non-steady state conditions such as competition or acute clinical pathologies. If so, CE could serve as a valuable tool for dynamically monitoring health status in a wide range of non-stationary systems. PMID- 19292506 TI - A lumped model for blood flow and pressure in the systemic arteries based on an approximate velocity profile function. AB - Previously, by assuming a viscous dominated flow in the boundary layer and an inertia dominated flow in the vessel core, a velocity profile function for a 1D wave propagation model was derived. Because the time dependent shape of the velocity profile in this boundary layer model depends on the size of the inviscid core and the boundary layer, and thus on the Womersley number, it differs along the arterial tree. In this study we evaluated a lumped model for a vessel segment in which the element configuration is based on physical phenomena described by the boundary layer model and for which all parameters have a physically based quantitative value dependent on the Womersley numbers. The proposed electrical analog consists of a Womersley number dependent resistor and an inductor arranged in parallel, representing the flow impedence in respectively the vessel core and the boundary layer, in series with a second resistor. After incorporating a capacitor representing the vessel compliance in this rigid tube model, the element configuration resembles the configuration of the four-element windkessel model. For arbitrary Womersley numbers the relative impedence of Womersley theory is approximated with high accuracy. In the limits for small and large Womersley numbers the relative impedences of the proposed lumped model correspond exactly to Womersley theory. PMID- 19292507 TI - Systemically modeling the dynamics of plasma insulin in subcutaneous injection of insulin analogues for type 1 diabetes. AB - Type 1 diabetics must inject exogenous insulin or insulin analogues one or more times daily. The timing and dosage of insulin administration have been a critical research area since the invention of insulin analogues. Several pharmacokinetical models have been proposed, and some are applied clinically in modeling various insulin therapies. However, their plasma insulin concentration must be computed separately from the models' output. Furthermore, minimal analytical study was performed in these existing models. We propose two systemic and simplified ordinary differential equation models to model the subcutaneous injection of rapid-acting insulin analogues and long-acting insulin analogues, respectively. Our models explicitly model the plasma insulin and hence have the advantage of computing the plasma insulin directly. The profiles of plasma insulin concentrations obtained from these two models are in good agreement with the experimental data. We also study the dynamics of insulin analogues, plasma insulin concentrations, and, in particular, the shape of the dynamics of plasma insulin concentrations. PMID- 19292508 TI - Feedback regulation in multistage cell lineages. AB - Studies of developing and self-renewing tissues have shown that differentiated cell types are typically specified through the actions of multistage cell lineages. Such lineages commonly include a stem cell and multiple progenitor (transit amplifying; TA) cell stages, which ultimately give rise to terminally differentiated (TD) cells. In several cases, self-renewal and differentiation of stem and progenitor cells within such lineages have been shown to be under feedback regulation. Together, the existence of multiple cell stages within a lineage and complex feedback regulation are thought to confer upon a tissue the ability to autoregulate development and regeneration, in terms of both cell number (total tissue volume) and cell identity (the proportions of different cell types, especially TD cells, within the tissue). In this paper, we model neurogenesis in the olfactory epithelium (OE) of the mouse, a system in which the lineage stages and mediators of feedback regulation that govern the generation of terminally differentiated olfactory receptor neurons (ORNs) have been the subject of much experimental work. Here we report on the existence and uniqueness of steady states in this system, as well as local and global stability of these steady states. In particular, we identify parameter conditions for the stability of the system when negative feedback loops are represented either as Hill functions, or in more general terms. Our results suggest that two factors -- autoregulation of the proliferation of transit amplifying (TA) progenitor cells, and a low death rate of TD cells -- enhance the stability of this system. PMID- 19292509 TI - Computation of traveling wave fronts for a nonlinear diffusion-advection model. AB - This paper utilizes a nonlinear reaction-diffusion-advection model for describing the spatiotemporal evolution of bacterial growth. The traveling wave solutions of the corresponding system of partial differential equations are analyzed. Using two methods, we then find such solutions numerically. One of the methods involves the traveling wave equations and solving an initial-value problem, which leads to accurate computations of the wave profiles and speeds. The second method is to construct time-dependent solutions by solving an initial-moving boundary-value problem for the PDE system, showing another approximation for such wave solutions. PMID- 19292510 TI - Estimation and identification of parameters in a lumped cerebrovascular model. AB - This study shows how sensitivity analysis and subset selection can be employed in a cardiovascular model to estimate total systemic resistance, cerebrovascular resistance, arterial compliance, and time for peak systolic ventricular pressure for healthy young and elderly subjects. These quantities are parameters in a simple lumped parameter model that predicts pressure and flow in the systemic circulation. The model is combined with experimental measurements of blood flow velocity from the middle cerebral artery and arterial finger blood pressure. To estimate the model parameters we use nonlinear optimization combined with sensitivity analysis and subset selection. Sensitivity analysis allows us to rank model parameters from the most to the least sensitive with respect to the output states (cerebral blood flow velocity and arterial blood pressure). Subset selection allows us to identify a set of independent candidate parameters that can be estimated given limited data. Analyses of output from both methods allow us to identify five independent sensitive parameters that can be estimated given the data. Results show that with the advance of age total systemic and cerebral resistances increase, that time for peak systolic ventricular pressure is increases, and that arterial compliance is reduced. Thus, the method discussed in this study provides a new methodology to extract clinical markers that cannot easily be assessed noninvasively. PMID- 19292511 TI - An in vivo intermediate filament assembly model. AB - A model is developed to study the in vivo intermediate filament organization in terms of repartition between four different structural states: soluble proteins, particles, short, and long filaments. An analysis is conducted, showing that the system has a unique, globally asymptotically stable equilibrium. By means of sensitivity analysis, the influence of parameters on the system is studied. It is shown that, in agreement with biological observations, posttranslational modifications of intermediate filament proteins resulting in filament solubilization are the main regulators of the intermediate filament organization. A high signalling-dependent solubilization of filaments favours the intermediate filament aggregation in particles. PMID- 19292512 TI - Modeling HIV outbreaks: the male to female prevalence ratio in the core population. AB - What affects the ratio of infected men to infected women in the core population in a heterosexual HIV epidemic? Hethcote and Yorke [5] introduced the term "core" initially to loosely describe the collection of individuals having the most unprotected sex partners. We study the early epidemic during the exponential growth phase and focus on the core group because most infected people were infected by people in the core. We argue that in the early outbreak phase of an epidemic, there is an identity, which we call the "outbreak equation." It relates three ratios that describe the core men versus the core women, namely, the ratio E of numbers of all core men to all core women, the ratio C of numbers of infected core men to core women, and the ratio M of the infectiousness of a typical core man to that of a typical core woman. Then the relationship between the ratios is E=MC(2) in the early outbreak phase. We investigate two very different scenarios, one in which there are two times as many core men as core women (E=2) and the other in which core men equal core women (E=1). In the first case, the HIV epidemic grows at a much faster rate. We conclude that if the female core group was larger, that is, if more women in the total population were promiscuous (or if fewer men were promiscuous) then the HIV epidemic would grow more slowly. PMID- 19292513 TI - The dynamics of a simple Laissez-Faire model with two predators. AB - In this paper, we study the dynamics of a laissez-faire predator--prey model with both a specialist and a generalist predator. We analyze the stabilities of equilibria by performing linearized stability analyses. We then reexamine the stability of the equilibrium where the prey and predator coexist by constructing a Lyapunov function. If we hold the generalist predator population constant, treating it as a bifurcation parameter, we show that our model can possess multiple (up to three) limit cycles that surround an equilibrium in the interior of the first quadrant. Our model shows rich dynamics including fold, transcritical, pitchfork, Hopf, cyclic-fold, and Bautin bifurcations as well as heteroclinic connections. If we instead vary the generalist predator population slowly across bifurcations, the model exhibits bursting behavior as it alternates between a repetitive spiking phase and a quiescent phase. PMID- 19292514 TI - Solution of the Michaelis-Menten equation using the decomposition method. AB - We present a low-order recursive solution to the Michaelis-Menten equation using the decomposition method. This solution is algebraic in nature and provides a simpler alternative to numerical approaches such as differential equation evaluation and root-solving techniques that are currently used to compute substrate concentration in the Michaelis-Menten equation. A detailed characterization of the errors in substrate concentrations computed from decomposition, Runge-Kutta, and bisection methods over a wide range of s(0) : K(m) values was made by comparing them with highly accurate solutions obtained using the Lambert W function. Our results indicated that solutions obtained from the decomposition method were usually more accurate than those from the corresponding classical Runge-Kutta methods. Moreover, these solutions required significantly fewer computations than the root-solving method. Specifically, when the stepsize was 0.1% of the total time interval, the computed substrate concentrations using the decomposition method were characterized by accuracies on the order of 10(-8) or better. The algebraic nature of the decomposition solution and its relatively high accuracy make this approach an attractive candidate for computing substrate concentration in the Michaelis-Menten equation. PMID- 19292515 TI - Dynamic effects and information quantifiers of statistical memory of MEG's signals at photosensitive epilepsy. AB - The time series analysis of magnetoencephalographic (MEG) signals is very important both for basic brain research and for medical diagnosis and treatment. Here we discuss the crucial role of statistical memory effects (ME) in human brain functioning with photosensitive epilepsy (PSE). We study two independent statistical memory quantifiers that reflect the dynamical characteristics of neuromagnetic brain responses on a flickering stimulus of different colored combinations from a group of control subjects, which are contrasted with those from a patient with PSE. We analyze the frequency dependence of two memory measures for the neuromagnetic signals. The strong memory and the accompanying transition to a regular and robust regime of the signals' chaotic behavior in the separate areas are characteristic for a patient with PSE. This particularly interesting observation most likely identifies the regions of the protective mechanism in a human organism against occurrence of PSE. PMID- 19292516 TI - The effect of global travel on the spread of SARS. PMID- 19292517 TI - Depletion effects in a mixture of hard and attractive colloids. AB - Monte Carlo simulation and theory were used to study the potential of mean force (PMF) between a pair of big colloidal (solute) particles suspended in a sea of smaller particles (solvent) interacting via Baxter's sticky hard sphere (SHS) potential. Simulation results were obtained by applying a special simulation technique developed for sampling the hard sphere collision force, while the theoretical predictions were calculated from the analytic solution of the Percus Yevick/Ornstein-Zernike integral equation for spatial correlations in a two component mixture at vanishing solute concentration. Both theory and simulation revealed oscillations of the solute-solute PMF with a period equal to the diameter of the solvent molecules. Further, the attractive PMF between solute particles in the SHS fluid decays slower than in a hard sphere solvent. Upon increasing the strength of attraction (stickiness) between the molecules of solvent, these oscillations gradually disappear, the PMF becoming long ranged and attractive at all separations. PMID- 19292518 TI - Energy partitioning for generalized product functions: the interference contribution to the energy of generalized valence bond and spin coupled wave functions. AB - The main driving force for the formation of the covalent bond is the quantum mechanical interference effect among one-electron states, as has been suggested in several works by the use of partition schemes to calculate the interference contributions to the energy. However, due to some difficulties associated with the original approaches, calculations were only carried out for a few, mostly diatomic molecules. In this work, we propose a general approach of partitioning based on generalized product functions with generalized valence bond at the perfect pairing approximation and spin-coupled groups, which should allow the investigation of a broader array of molecules, and hopefully, shed light on the nature of the chemical bond in molecules with unusual chemical features. Among other things, this approach lends itself naturally to the investigation of interference in individual bonds or groups of bonds in a molecule. PMID- 19292519 TI - Solid-liquid coexistence of polydisperse fluids via simulation. AB - We describe a simulation method for the accurate study of the equilibrium freezing properties of polydisperse fluids under the experimentally relevant condition of fixed polydispersity. The approach is based on the phase switch Monte Carlo method of Wilding and Bruce [Phys. Rev. Lett. 85, 5138 (2000)]. This we have generalized to deal with particle size polydispersity by incorporating updates which alter the diameter sigma of a particle, under the control of a distribution of chemical potential differences mu(sigma). Within the resulting isobaric semi-grand-canonical ensemble, we detail how to adapt mu(sigma) and the applied pressure such as to study coexistence, while ensuring that the ensemble averaged density distribution rho(sigma) matches a fixed functional form. Results are presented for the effects of small degrees of polydispersity on the solid liquid transition of soft spheres. PMID- 19292520 TI - A modified ansatz for explicitly correlated coupled-cluster wave functions that is suitable for response theory. AB - A modified ansatz for explicitly correlated coupled-cluster wave functions with a single correlation factor is set forward. It is based on the fixed amplitude ansatz of Ten-no [Chem. Phys. Lett. 398, 56 (2004)] to which an extra term is added that allows for the explicitly correlated description of singly excited configurations. The new approach has been implemented for coupled-cluster singles and doubles with the aid of automated techniques. Numerical results are presented for vertical excitation energies, and ground and excited state equilibrium distances and harmonic frequencies of diatomics. The new approach is shown to provide a nearly unbiased description of ground and predominantly singly exited states, and the improvements seen for explicitly correlated treatments of ground states, as compared to conventional orbital expansions, carry over to excited states. In addition, a correction for contributions from one-electron terms-which are not improved by the correlation factor-is suggested, again with focus on applicability to a response formalism. PMID- 19292521 TI - Improving the accuracy of the nonlocal van der Waals density functional with minimal empiricism. AB - The nonlocal van der Waals density functional (vdW-DF) captures the essential physics of the dispersion interaction not only in the asymptotic regime but also for a general case of overlapping fragment densities. A balanced treatment of other energetic contributions, such as exchange, is crucial if we aim for accurate description of various properties of weakly bound systems. In this paper, the vdW-DF correlation functional is modified to make it better compatible with accurate exchange functionals. We suggest a slightly simplified construction of the nonlocal correlation, yielding more accurate asymptotic C(6) coefficients. We also derive a gradient correction, containing a parameter that can be adjusted to suit an exchange functional of choice. We devise a particularly apt combination of exchange and correlation terms, which satisfies many important constraints and performs well for our benchmark tests. PMID- 19292522 TI - Removing systematic errors in interionic potentials of mean force computed in molecular simulations using reaction-field-based electrostatics. AB - The performance of reaction-field methods to treat electrostatic interactions is tested in simulations of ions solvated in water. The potential of mean force between sodium chloride pair of ions and between side chains of lysine and aspartate are computed using umbrella sampling and molecular dynamics simulations. It is found that in comparison with lattice sum calculations, the charge-group-based approaches to reaction-field treatments produce a large error in the association energy of the ions that exhibits strong systematic dependence on the size of the simulation box. The atom-based implementation of the reaction field is seen to (i) improve the overall quality of the potential of mean force and (ii) remove the dependence on the size of the simulation box. It is suggested that the atom-based truncation be used in reaction-field simulations of mixed media. PMID- 19292523 TI - Accuracies of the empirical theories of the escape probability based on Eigen model and Braun model compared with the exact extension of Onsager theory. AB - This paper deals with the exact extension of the original Onsager theory of the escape probability to the case of finite recombination rate at nonzero reaction radius. The empirical theories based on the Eigen model and the Braun model, which are applicable in the absence and presence of an external electric field, respectively, are based on a wrong assumption that both recombination and separation processes in geminate recombination follow exponential kinetics. The accuracies of the empirical theories are examined against the exact extension of the Onsager theory. The Eigen model gives the escape probability in the absence of an electric field, which is different by a factor of 3 from the exact one. We have shown that this difference can be removed by operationally redefining the volume occupied by the dissociating partner before dissociation, which appears in the Eigen model as a parameter. The Braun model gives the escape probability in the presence of an electric field, which is significantly different from the exact one over the whole range of electric fields. Appropriate modification of the original Braun model removes the discrepancy at zero or low electric fields, but it does not affect the discrepancy at high electric fields. In all the above theories it is assumed that recombination takes place only at the reaction radius. The escape probability in the case when recombination takes place over a range of distances is also calculated and compared with that in the case of recombination only at the reaction radius. PMID- 19292524 TI - Bounding the electrostatic free energies associated with linear continuum models of molecular solvation. AB - The importance of electrostatic interactions in molecular biology has driven extensive research toward the development of accurate and efficient theoretical and computational models. Linear continuum electrostatic theory has been surprisingly successful, but the computational costs associated with solving the associated partial differential equations (PDEs) preclude the theory's use in most dynamical simulations. Modern generalized-Born models for electrostatics can reproduce PDE-based calculations to within a few percent and are extremely computationally efficient but do not always faithfully reproduce interactions between chemical groups. Recent work has shown that a boundary-integral-equation formulation of the PDE problem leads naturally to a new approach called boundary integral-based electrostatics estimation (BIBEE) to approximate electrostatic interactions. In the present paper, we prove that the BIBEE method can be used to rigorously bound the actual continuum-theory electrostatic free energy. The bounds are validated using a set of more than 600 proteins. Detailed numerical results are presented for structures of the peptide met-enkephalin taken from a molecular-dynamics simulation. These bounds, in combination with our demonstration that the BIBEE methods accurately reproduce pairwise interactions, suggest a new approach toward building a highly accurate yet computationally tractable electrostatic model. PMID- 19292525 TI - Topology of the quantum control landscape for observables. AB - A broad class of quantum control problems entails optimizing the expectation value of an observable operator through tailored unitary propagation of the system density matrix. Such optimization processes can be viewed as a directed search over a quantum control landscape. The attainment of the global extrema of this landscape is the goal of quantum control. Local optima will generally exist, and their enumeration is shown to scale factorially with the system's effective Hilbert space dimension. A Hessian analysis reveals that these local optima have saddlepoint topology and cannot behave as suboptimal extrema traps. The implications of the landscape topology for practical quantum control efforts are discussed, including in the context of nonideal operating conditions. PMID- 19292526 TI - Enhanced conformational sampling of nucleic acids by a new Hamiltonian replica exchange molecular dynamics approach. AB - Although molecular dynamics (MD) simulations have been applied frequently to study flexible molecules, the sampling of conformational states separated by barriers is limited due to currently possible simulation time scales. Replica exchange (Rex)MD simulations that allow for exchanges between simulations performed at different temperatures (T-RexMD) can achieve improved conformational sampling. However, in the case of T-RexMD the computational demand grows rapidly with system size. A Hamiltonian RexMD method that specifically enhances coupled dihedral angle transitions has been developed. The method employs added biasing potentials as replica parameters that destabilize available dihedral substates and was applied to study coupled dihedral transitions in nucleic acid molecules. The biasing potentials can be either fixed at the beginning of the simulation or optimized during an equilibration phase. The method was extensively tested and compared to conventional MD simulations and T-RexMD simulations on an adenine dinucleotide system and on a DNA abasic site. The biasing potential RexMD method showed improved sampling of conformational substates compared to conventional MD simulations similar to T-RexMD simulations but at a fraction of the computational demand. It is well suited to study systematically the fine structure and dynamics of large nucleic acids under realistic conditions including explicit solvent and ions and can be easily extended to other types of molecules. PMID- 19292527 TI - The calculation of adiabatic-connection curves from full configuration interaction densities: two-electron systems. AB - The Lieb formulation of density-functional theory is briefly reviewed and its straightforward generalization to arbitrary electron-electron interaction strengths discussed, leading to the introduction of density-fixed and potential fixed adiabatic connections. An iterative scheme for the calculation of the Lieb functionals under the appropriate constraints is outlined following the direct optimization approach of Wu and Yang [J. Chem. Phys. 118, 2498 (2003)]. First- and second-order optimization schemes for the calculation of accurate adiabatic connection integrands are investigated and compared; the latter is preferred both in terms of computational efficiency and accuracy. The scheme is applicable to systems of any number of electrons. However, to determine the accuracy that may be achieved, the present work focuses on two-electron systems for which a number of simplifications may be exploited. The procedure is applied to the helium isoelectronic series and the H(2) molecule. The resulting adiabatic-connection curves yield the full configuration-interaction exchange-correlation energies extrapolated to the basis-set limit. The relationship between the Kohn-Sham and natural orbitals as functions of the electron-electron interaction strength is explored in detail for H(2). The accuracy with which the exchange-correlation contributions to the modified local potential can be determined is discussed. The new accurate adiabatic-connection curves are then compared with some recently investigated approximate forms calculated using accurate full configuration interaction input data. This study demonstrates that the adiabatic-connection integrand may be determined accurately and efficiently, providing important insights into the link between the Kohn-Sham and traditional quantum-chemical treatments of the exchange-correlation problem in electronic-structure theory. PMID- 19292528 TI - Characterization of the HSiN_HNSi system in its electronic ground state. AB - The electronic ground states (X (1)Sigma(+)) of HSiN, HNSi, and the transition state connecting the two isomers were systematically studied using configuration interaction with single and double (CISD) excitations, coupled cluster with single and double (CCSD) excitations, CCSD with perturbative triple corrections [CCSD(T)], multireference complete active space self-consistent field (CASSCF), and internally contracted multireference configuration interaction (ICMRCI) methods. The correlation-consistent polarized valence (cc-pVXZ), augmented correlation-consistent polarized valence (aug-cc-pVXZ) (X=T,Q,5), correlation consistent polarized core-valence (cc-pCVYZ), and augmented correlation consistent polarized core-valence (aug-cc-pCVYZ) (Y=T,Q) basis sets were used. Via focal point analyses, we confirmed the HNSi isomer as the global minimum on the ground state HSiN_HNSi zero-point vibrational energy corrected surface and is predicted to lie 64.7 kcal mol(-1) (22 640 cm(-1), 2.81 eV) below the HSiN isomer. The barrier height for the forward isomerization reaction (HSiN-->HNSi) is predicted to be 9.7 kcal mol(-1), while the barrier height for the reverse process (HNSi-->HSiN) is determined to be 74.4 kcal mol(-1). The dipole moments of the HSiN and HNSi isomers are predicted to be 4.36 and 0.26 D, respectively. The theoretical vibrational isotopic shifts for the HSiN/DSiN and HNSi/DNSi isotopomers are in strong agreement with the available experimental values. The dissociation energy for HSiN [HSiN(X (1)Sigma(+))-->H((2)S)+SiN(X (2)Sigma(+))] is predicted to be D(0)=59.6 kcal mol(-1), whereas the dissociation energy for HNSi [HNSi(X (1)Sigma(+))-->H((2)S)+NSi(X (2)Sigma(+))] is predicted to be D(0)=125.0 kcal mol(-1) at the CCSD(T)/aug-cc-pCVQZ level of theory. Anharmonic vibrational frequencies computed using second order vibrational perturbation theory are in good agreement with available matrix isolation experimental data for both HSiN and HNSi isomers root mean squared derivation (RMSD=9 cm(-1)). PMID- 19292529 TI - Photodissociation of the linear Ar-I2 van der Waals complex: velocity-map imaging of the I2 fragment. AB - Photodissociation of the Ar-I(2) 1:1 linear van der Waals complex is studied over the 490-520 nm region using the velocity-map imaging technique. Molecular iodine, and both the T-shaped and linear Ar-I(2)(X,v(")=0) ground-state complexes absorb strongly in this range, and these transitions access both the bound and dissociative regions of the I(2)(B) state. We measure the angle-speed distribution of vibrationally excited I(2)(B,v(')) state products by resonant 1+1 ionization via the E and f ion-pair states, forming I(2) (+), which is imaged under velocity-mapping conditions. The images show a strong angular anisotropy, which is the same for all excitation energies, spanning from the bound region to above the molecular dissociation limit. The observed angular anisotropy of the I(2) fragments is consistent with a direct dissociation of linear Ar-I(2) complexes promoted to the inner repulsive potential wall of the Ar+I(2)(B,v(')) potential energy surface. PMID- 19292530 TI - The water-nitric oxide intermolecular potential-energy surface revisited. AB - The two lowest energy intermolecular potential-energy surfaces (IPESs) of the water-nitric oxide complex are evaluated using the spin-restricted coupled cluster R-CCSD(T) model and the augmented correlation-consistent polarized valence triple-zeta basis set extended with a set of the 3s3p2d1f1g midbond functions. A detailed characterization of the IPESs for both the (2)A(') and (2)A(") electronic states in the C(s)-symmetry configurations of the complex is performed. The global minimum for the (2)A(') state represented by the lowest energy of -461.8 cm(-1) is deeper than the global minimum in the (2)A(") state with an energy of -435.2 cm(-1). To explore the physics of the interaction an open-shell implementation of the symmetry-adapted perturbation theory is employed and the results are analyzed as a function of the intermolecular parameters. The electrostatic term shows the strongest geometric anisotropy, while the exchange, induction, and dispersion contributions mostly depend on the intermolecular distance. The energy separation between the (2)A(') and (2)A(") states is largely dominated by electrostatic contribution for long intermolecular distances. In the region of short intermolecular distances the exchange part is as important as the electrostatic one and the induction and dispersion effects are also substantial. PMID- 19292531 TI - Intra- and intermolecular electrostatic interactions and their significance for the structure, acidity, and tautomerization behavior of trinitromethane. AB - We have addressed several interesting issues related to trinitromethane: the propellerlike arrangement of its nitro groups, its lower-than-predicted (although still high) acidity, its aci tautomerization, and the absence of expected very short C-HO intermolecular bridges in the crystal. A combination of crystallographic and computed data was used in our analysis. The structural features mentioned and the anomalous acidity can be attributed to intra- and/or intermolecular electrostatic interactions. Decomposition via C-NO(2) bond scission may occur before aci tautomerization can take place. PMID- 19292532 TI - Effects of pi-stacking interactions on the near carbon K-edge x-ray absorption fine structure: a theoretical study of the ethylene pentamer and the phthalocyanine dimer. AB - X-ray absorption spectra have been determined for ethylene and free base phthalocyanine at the carbon K-edge with use of the complex polarization propagator method combined with Kohn-Sham density functional theory and the Coulomb attenuated method B3LYP exchange-correlation functional. Apart from isolated molecules, the study includes pi-stacked systems of the phthalocyanine dimer and the ethylene dimer, trimer, tetramer, and pentamer. For ethylene, pi stacking involves a reduction in transition energy of the valence pi( *)-band by some 70 meV and large spectral changes (regarding also shape and intensity) of the Rydberg bands. For phthalocyanine, there are large spectral changes in the entire valence pi( *)-part of the spectrum. PMID- 19292533 TI - On the dissociation of I2 by O2(a1Delta): Pathways involving the excited species I2(A'3Pi2u,A3Pi(1u)), I2(X1sigma,upsilon), and O2(a1Delta,upsilon). AB - Kinetic studies were carried out to explore the role of the excited species I(2)(A(') (3)Pi(2u),A (3)Pi(1u)), I(2)(X (1) summation operator,upsilon), and O(2)(a (1)Delta,upsilon) in the dissociation of I(2) by singlet oxygen. A flow tube apparatus that utilized a chemical singlet oxygen generator was used to measure the I(2) dissociation rate in O(2)(a (1)Delta)/I(2) mixtures. Vibrationally excited I(2)(X) is thought to be a significant intermediate in the dissociation process. Excitation probabilities (gamma(upsilon)) for population of the upsilonth I(2)(X) vibrational level in the reaction I(2)(X)+I((2)P(1/2))- >I(2)(X,upsilon>10)+I((2)P(3/2)) were estimated based on a comparison of calculated populations with experimentally determined values. Satisfactory agreement with the experimental data [Barnault et al., J. Phys. IV 1, C7/647 (1991)] was achieved for total excitation probabilities partitioned in two ranges, such that Gamma(25/=1%) the excited states are populated in the chain stage by collisions of I(2)(X,15 or =34 weeks and birth weight > or =2500 g. RESULTS: The average SpO(2) for all children was 98.0% (SD 1.4). SpO(2) was negatively related to birth weight and ranged from 98.2% (1.4) in children with birth weight 2750 2999 g to 97.6% (1.3) in children weighing >4500 g (P<0.001). No relationship exists between SpO(2) and gestational age or gender when correcting for birth weight. SpO(2) was independently related to mode of delivery, with higher levels in children born by cesarean section (98.3%; 1.3) than by vaginal delivery (98.0; 1.4; P<0.001). There was no relationship between SpO(2) and the interval from delivery to measurement. CONCLUSIONS: In healthy newborns, levels of SpO(2) measured between 2 and 24 h of life are negatively related to birth weight and related to mode of delivery. However, the variation was within a small range and probably has few implications for the routine use of SpO(2) in newborns. PMID- 19292586 TI - Severe neonatal hyporegenerative anemia due to anti-Vw (anti-MNS9) alloantibody. PMID- 19292587 TI - Determination of fetal heart rate reactivity from a single 20-min window of non stress testing in compromised fetuses. AB - AIMS: To shorten the analysis time needed for non-stress testing (NST) without decreasing efficacy in compromised fetuses. METHODS: We selected 80 cases with a 5-min Apgar score <7 as a study group and 259 cases with a 5-min Apgar score >/=9 as a control group. We applied four different criteria (A, B, C, and D) to each study and control group for the first 20-min window of NST data to evaluate reactivity. Criteria A, B, and C consisted of conventional reactivity criteria according to amplitude (15 or 10 beats per minute), duration (15 or 10 s) and weeks of gestation (/=32), and criteria D combined criteria C with approximate entropy (ApEn). RESULTS: The sensitivity of criteria D (91.25%) was greater than the other three criteria (P<0.0001). The specificities of criteria C (96.14%) and D (99.23%) were also higher than criteria A and B (P<0.0001). The positive and negative predictive value of criteria D were better than that of criteria C (97.33 vs. 83.87, P=0.0066) and (97.35 vs. 89.89, P=0.0004), respectively. CONCLUSION: Adding ApEn to the conventional criteria for reactivity shortened NST analysis time without decreasing efficacy, facilitating a decision of reactivity within a single 20-min NST window. PMID- 19292588 TI - The effect of bilateral pelvic neurectomy on uterine and abdominal electrical and pressure activity, as measured by telemetry in conscious, unrestrained pregnant rats. AB - AIMS: Bilateral pelvic neurectomy (BPN) interferes with delivery in rats. This work measures the effect of BPN on uterine and abdominal electromyography (EMG) and pressure during gestation and delivery. METHODS: Uterine EMG and intra uterine pressure (IUP), as well as abdominal muscle EMG and intra-abdominal pressure (IAP), were recorded on days 19-24 in rats. Mean values were determined for the peak frequency of the power density spectrum (PDS) of the uterine and abdominal EMG "bursts," the integral of the abdominal EMG power spectrum (IPS), and the area under the IUP and IAP curves (AUIUP and AUIAP). RESULTS: Uterine EMG power density spectrum peak frequency and AUIUP were low during days 19-21 in all rats, and on days 22-23 increased sharply and significantly in both groups. Before delivery, sham rat recordings showed sudden sharp increases in AUIAP, AUIUP and IPS as "pushing" was observed. Pushing was not observed or recorded in laboring BPN rats and they did not deliver normally. CONCLUSIONS: A functional pelvic nerve is required for normal labor and delivery in rats. BPN inhibits abdominal EMG and contractions necessary to expel the fetus. PMID- 19292589 TI - Parental mental health and child development from six to thirty-six months in a birth cohort study in Taiwan. AB - This study investigated the reciprocity between parental mental health and the different stages of child development at 6, 18, and 36 months. As the pilot of a birth cohort study, this study comprised 2048 children and their parents who were randomly selected and invited to participate. The development of these children and the mental health of their parents were followed at 6, 18, and 36 months postpartum. Child development was assessed using the Taiwan Birth Cohort Study instrument, and parental health was assessed using the Taiwanese version of the 36-Item Short Form Health Survey. Complete responses for all three stages were received from 844 families. Our results showed that parental mental health had a direct effect on language and social development; however, this effect did not become significant until 36 months. The reciprocity between child development and parental mental health and proper intervention are vital. PMID- 19292590 TI - Predictors of loss of virologic response in subjects who simplified to lopinavir/ritonavir monotherapy from lopinavir/ritonavir plus zidovudine/lamivudine. AB - Previous studies have demonstrated that lopinavir/ritonavir monotherapy maintained plasma HIV-1 RNA suppression in a large proportion of antiretroviral naive subjects. However, more subjects receiving lopinavir/ritonavir monotherapy experienced confirmed virologic rebound >50 copies/ml compared to a standard three-drug HAART regimen. In this study, we sought to determine the factors associated with maintenance of virologic suppression in subjects receiving lopinavir/ritonavir monotherapy. Antiretroviral-naive HIV-1-infected volunteers were randomized 2:1 to initiate a lopinavir/ritonavir-based combination regimen followed by simplification to lopinavir/ritonavir monotherapy or an efavirenz based triple combination therapy and followed for 96 weeks. Potential predictors of time to loss of virologic response included baseline demographics, baseline HIV-1 RNA levels, baseline CD4(+) T cell counts, adherence as determined by 4-day subject recall, duration of HIV-1 RNA <50 copies/ml prior to simplification, and lopinavir concentrations. By the Cox proportional hazards model, higher reported adherence levels and higher baseline CD4(+) T cell counts were associated with a greater likelihood of maintaining virologic suppression while receiving lopinavir/ritonavir monotherapy. Lopinavir concentrations, including trough concentrations, were not significantly associated with virologic outcomes. This analysis suggests that adherence and higher baseline CD4(+) T cell counts may help to predict who will sustain virologic suppression with lopinavir/ritonavir monotherapy. The data also suggest that measuring lopinavir concentrations is not useful in predicting virologic response in these patients. PMID- 19292591 TI - Re: "Enhancement of HIV infection by cellulose sulfate," by Tao et al. PMID- 19292592 TI - Unreliable real-time PCR analysis of human endogenous retrovirus-W (HERV-W) RNA expression and DNA copy number in multiple sclerosis. PMID- 19292594 TI - Virological suppression reduces clinical progression in patients with multiclass resistant HIV type 1. AB - The virological and immunological outcomes in patients carrying multiclass resistant HIV-1, their predictors, and their impact on disease progression were investigated. Antiretroviral-experienced patients carrying at least one primary resistance mutation (IAS-USA 2006) to two to three classes of antiretroviral drugs were analyzed for achieving an HIV-1 RNA <50 copies/ml, a CD4 count increase of >200 cells/microl from baseline, and progression to an AIDS-defining event or death. Survival analysis was performed using the Kaplan-Meier estimates and predictors of different outcomes were analyzed using Cox's regression models. A total of 236 patients were identified. Of these 73% reached HIV-1 RNA <50 copies/ml. Higher genotypic sensitivity score of the salvage regimen, lower viral load, and more recent calendar year at genotyping were independently associated with virological response. Immunological response (58%) was predicted by a more recent calendar year, the achievement of an undetectable viral load, and higher CD4 counts at genotyping. Thirty-three patients showed clinical progression: achieving HIV-1 RNA <50 copies/ml predicted AIDS-free survival, independently from other significant cofactors. In individuals with multiclass-resistant HIV-1, virological suppression and immunological recovery are becoming more easily accessible with more recent therapies. The achievement of virological suppression is a strong predictor of reduced clinical progression. PMID- 19292593 TI - Oligomer-specific conformations of the human immunodeficiency virus (HIV-1) gp41 envelope glycoprotein ectodomain recognized by human monoclonal antibodies. AB - Trimerization of the human immunodeficiency virus (HIV-1) envelope glycoproteins is mediated by the ectodomain of the gp41 transmembrane glycoprotein. Here we investigate oligomer-specific conformations of gp41 by using monoclonal antibodies (MAbs) from HIV-1-infected humans. Human MAbs directed against the cluster I region of gp41 recognized trimeric, dimeric, and monomeric forms of soluble envelope glycoproteins; thus, the integrity of the cluster I epitopes is minimally affected by the oligomeric state. In contrast, human MAbs to the cluster II region were all oligomers specific. One cluster II MAb, 126-6, recognized exclusively the trimeric form of envelope glycoproteins, whereas the others recognized both trimeric and dimeric forms. Thus, a distinct trimer specific conformation exists in the cluster II region of gp41. Analysis of soluble envelope glycoprotein mutants revealed that gp41 sequences immediately N terminal to isoleucine 646 contribute to the formation of both the trimer and the trimer-specific conformational epitope. PMID- 19292595 TI - Human immunodeficiency virus (HIV) antigens and RNA in HIV-seronegative women with cervical intraepithelial neoplasia. AB - While investigating whether proteins retrieved by cervicovaginal lavages (CVL) from women with cervical intraepithelial neoplasia (CIN) might correlate with risk of progression to invasive cervical cancer, we unexpectedly identified HIV gag and env glycoprotein in CVL from women with HIV-negative serology. HIV antigens were consistently identified by mass spectrometry (MS) in CVL from 4 women but were absent in CVL from the remaining 16 women. HIV serologies of all 20 patients were negative for both HIV-1 and HIV-2 antibodies. To validate the unexpected MS findings we performed Western blot (WB) and immunoaffinity chromatography (IC) analysis of CVL for HIV proteins, viral load assays of paired CVL and blood samples, and immunohistochemical HIV p24 expression in cervical biopsy specimens. WB analysis of CVL for prostate-specific antigen (PSA) was performed to exclude semen contamination as the source of HIV proteins. WB and IC results demonstrated the presence of HIV-1 gp41 and p24 antigens in four CVL that were identified by MS to have the HIV proteins. Despite negative serology, HIV RNA in CVL and HIV p24 in cervix biopsies were detected in patients with HIV antigen-positive CVL. HIV p24-positive CVL were PSA negative. All 20 subjects remained HIV seronegative throughout the study. Women with HIV proteins and RNA were comparatively older. Our findings suggest that CVL HIV proteins in women with CIN could be markers for unrecognized HIV exposure or subclinical infection. Proteomic screening of cervical secretions may be useful in identifying seronegative women exposed to HIV and/or at risk for AIDS. PMID- 19292596 TI - A comparison of interpretation by three different HIV type 1 genotypic drug resistance algorithms using sequences from non-clade B HIV type 1 strains. AB - The advent of affordable ART has benefited HIV-infected individuals. Prospective studies have shown that the availability of drug resistance reports for infected individuals has allowed more effective regimens to be prescribed as compared to a control group whose physicians had no access to drug resistance reports. There is a paucity of information on the performance of genotypic algorithms on non-clade B HIV-1 strains, especially clade C. In this study the results obtained on submission of HIV-1 RT and PR sequences of non-clade B strains to the Stanford University HIV drug resistance database (SHDB) were compared to the results obtained from Geno2Pheno (G2P) and DR_Seqan (DS). For the study, we took samples from 93 treatment-naive individuals and 21 samples from 19 infected individuals showing detectable viral load while on ART. There were discrepancies in the clade identification results obtained from the SHDB and G2P databases. This feature was not available in DS. The mean observed concordance between SHDB and G2P was 85.6% while between SHDB and DC it was 37%. When the level of concordance was determined based on exposure to ART, the G2P was found to have a better level of concordance (76.8%) to SHDB as compared to SHDB versus DS (36%). We do not have phenotypic data for the strains included in this study and hence we are not in a position to assign a particular algorithm as being superior. These results also show a possible need for a subtype-specific algorithm for interpretation of HIV-1 genotypic drug resistance. PMID- 19292598 TI - Biochemical properties, pharmacokinetics and pharmacological response of tiotropium in chronic obstructive pulmonary disease patients. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a progressive disease with increasing incidence and mortality. Tiotropium is an inhaled long-acting anti-cholinergic for the maintenance treatment of COPD. OBJECTIVE: To review biochemical and pharmacokinetic data on tiotropium and discuss in the context of tiotropium's efficacy and safety in COPD. METHODS: Review of previously done pharmacokinetic studies performed by the manufacturer of tiotropium. Data obtained through peer-reviewed publications and regulatory websites. RESULTS/CONCLUSIONS: The long duration of action with tiotropium is owing to prolonged, competitive binding to M(3) muscarinic receptors. Tiotropium is poorly absorbed following inhalation, which largely limits side effects. Metabolism of absorbed drug is minimal and excretion is largely through the kidneys. Tiotropium is efficacious and well tolerated by patients with COPD. PMID- 19292599 TI - New trends in transplantation: the use of Thymoglobulin. AB - BACKGROUND: Thymoglobulin (Genzyme Co., Cambridge, MA, USA), rabbit anti thymocyte globulin, is an immunosuppressive drug approved for the treatment of renal transplant rejection. OBJECTIVE: To review the pharmacology, toxicity and efficacy of Thymoglobulin in solid organ and stem cell transplantations. METHODS: Review of the literature, information from Genzyme and personal clinical trial experience serve as sources for this review. CONCLUSION: Thymoglobulin has helped to facilitate the growth of successful steroid sparing approaches to solid organ transplantation. Thymoglobulin is now incorporated into several stem cell transplantation approaches. Infection and second malignancies remain as concerns. Randomized trials are needed to compare efficacy with standard regimens. PMID- 19292600 TI - Allergic rhinitis in patients with asthma: the Swiss LARA (Link Allergic Rhinitis in Asthma) survey. AB - OBJECTIVE: To determine the characteristics of asthma (A) and allergic rhinitis (AR) among asthma patients in primary care practice. RESEARCH DESIGN AND METHODS: Primary care physicians, pulmonologists, and allergologists were asked to recruit consecutive asthma patients with or without allergic rhinitis from their daily practice. Cross-sectional data on symptoms, severity, treatment and impact on quality of life of A and AR were recorded and examined using descriptive statistics. Patients with and without AR were then compared. RESULTS: 1244 asthma patients were included by 211 physicians. Asthma was controlled in 19%, partially controlled in 27% and not controlled in 54%. Asthma treatment was generally based on inhaled corticosteroids (ICS) with or without long acting beta 2 agonists (78%). A leukotriene receptor antagonist (LTRA) was used by 46% of the patients. Overall, 950 (76%) asthma patients had AR (A + AR) and 294 (24%) did not (A - AR). Compared to patients with A - AR, A + AR patients were generally younger (mean age +/- standard deviation: 42 +/- 16 vs. 50 +/- 19 years, p < 0.001) and fewer used ICS (75% vs. 88%, p < 0.001). LTRA usage was similar in both groups (46% vs. 48%). Asthma was uncontrolled in 53% of A + AR and 57% of A - AR patients. Allergic rhinitis was treated with a mean of 1.9 specific AR medications: antihistamines (77%), nasal steroids (66%) and/or vasoconstrictors (38%), and/or LTRA (42%). Rhinorrhoea, nasal obstruction, or nasal itching were the most frequently reported AR symptoms and the greatest reported degree of impairment was in daily activities/sports (55%). CONCLUSIONS: Allergic rhinitis was more common among younger asthma patients, increased the burden of symptoms and the need for additional medication but was associated with improved asthma control. However, most asthma patients remained suboptimally controlled regardl ess of concomitant AR. PMID- 19292601 TI - Bone health in diabetes: considerations for clinical management. AB - BACKGROUND: The metabolic and endocrine alterations of diabetes adversely affect bone quantity and/or quality and may increase fracture risk. SCOPE: A survey of the scientific literature on diabetes and bone cited on PubMed/MEDLINE and published in English from January 1970 to November 2008. FINDINGS: Subjects with type 1 diabetes have reduced bone mass and increased risk of fragility fracture, while those with type 2 diabetes, despite having normal or above-normal bone mineral density (BMD), are susceptible to low-trauma fractures, especially hip fractures. A recent meta-analysis, involving 836 000 subjects and 139 000 incident cases of fracture, found that type 2 diabetes was associated with significantly increased risks of non-vertebral (relative risk 1.2), hip (relative risk 1.7) and foot (relative risk 1.3) fracture. The association with hip fracture persisted after adjustment for age, physical activity and body weight, and was more pronounced in men and in those with long-standing diabetes. Insulin has an anabolic effect on bone, and the qualitatively different effects of type 1 and type 2 diabetes on bone mass are consistent with the opposing insulin secretory states (hypoinsulinaemia vs. hyperinsulinaemia). However, the existence of an elevated fracture risk in type 2 diabetes, despite the underlying hyperinsulinaemia, suggests the involvement of other potential pathogenic influences (e.g., hyperglycaemia, diabetic complications and lifestyle factors) on bone. Animal studies suggest that diabetic bone may be more fragile than non diabetic bone. Falls arising from diabetes-related comorbidities are another possible cause of low-trauma fracture. Clinical trial findings, supported by bone marker and bone density data, suggest that the oral antidiabetic agents metformin and glibenclamide significantly lower fracture risk, whereas the thiazolidinediones slightly increase fracture risk in postmenopausal women, but not in men, with type 2 diabetes. Recent preclinical studies have helped elucidate the mechanisms underlying the dynamics of bone remodelling, but more research is needed to improve outcomes for patients. CONCLUSIONS: Bone health is an important consideration in diabetes, and caution should be exercised in prescribing thiazolidinediones to postmenopausal women with low BMD and patients with prior fracture. PMID- 19292602 TI - Sleep disturbances in patients with chronic pain: effectively managing opioid analgesia to improve outcomes. AB - BACKGROUND: Many patients with chronic pain experience pain-related sleep disturbances, such as difficulty falling and staying asleep and less restful sleep. Evidence suggests that pain and sleep exist in a bidirectional relationship in which pain causes sleep disturbance and sleep disturbance intensifies pain. This association can impair a patient's daily function and decrease quality of life. Evidence suggests that patients with chronic pain can use opioid analgesics or other pain medications to control their pain and, in turn, improve some measures of sleep. This may include subjective sleep measures such as increased sleep time, and, as evidenced in recent studies, objective sleep measures such as sleep efficiency. SCOPE: The role of effective analgesia in the improvement of pain-related sleep disturbance is discussed herein, specifically the risks and benefits of opioid therapy for the treatment of patients with chronic pain and disturbed sleep. MEDLINE and PubMed searches were conducted to locate relevant studies dated from January 1975 to April 2008. English-only randomized controlled trials and nonrandomized studies were considered. FINDINGS: Numerous studies support the benefits of effective analgesia with opioid therapy on sleep. CONCLUSION: Pain control achieved with pharmacotherapy, specifically opioid therapy, may help to improve sleep in patients for which opioid therapy is appropriate. PMID- 19292603 TI - Signatures of a shadow biosphere. AB - Astrobiologists are aware that extraterrestrial life might differ from known life, and considerable thought has been given to possible signatures associated with weird forms of life on other planets. So far, however, very little attention has been paid to the possibility that our own planet might also host communities of weird life. If life arises readily in Earth-like conditions, as many astrobiologists contend, then it may well have formed many times on Earth itself, which raises the question whether one or more shadow biospheres have existed in the past or still exist today. In this paper, we discuss possible signatures of weird life and outline some simple strategies for seeking evidence of a shadow biosphere. PMID- 19292607 TI - Infection, allergy, and the protection from breastfeeding. PMID- 19292604 TI - Transduction of nonhuman primate brain with adeno-associated virus serotype 1: vector trafficking and immune response. AB - We used convection-enhanced delivery (CED) to characterize gene delivery mediated by adeno-associated virus type 1 (AAV1) by tracking expression of hrGFP (humanized green fluorescent protein from Renilla reniformis) into the striatum, basal forebrain, and corona radiata of monkey brain. Four cynomolgus monkeys received single infusions into corona radiata, putamen, and caudate. The other group (n = 4) received infusions into basal forebrain. Thirty days after infusion animals were killed and their brains were processed for immunohistochemical evaluation. Volumetric analysis of GFP-positive brain areas was performed. AAV1 hrGFP infusions resulted in approximately 550, 700, and 73 mm(3) coverage after infusion into corona radiata, striatum, and basal forebrain, respectively. Aside from targeted regions, other brain structures also showed GFP signal (internal and external globus pallidus, subthalamic nucleus), supporting the idea that AAV1 is actively trafficked to regions distal from the infusion site. In addition to neuronal transduction, a significant nonneuronal cell population was transduced by AAV1 vector; for example, oligodendrocytes in corona radiata and astrocytes in the striatum. We observed a strong humoral and cell-mediated response against AAV1-hrGFP in transduced monkeys irrespective of the anatomic location of the infusion, as evidenced by induction of circulating anti-AAV1 and anti-hrGFP antibodies, as well as infiltration of CD4(+) lymphocytes and upregulation of MHC II in regions infused with vector. We conclude that transduction of antigen presenting cells within the CNS is a likely cause of this response and that caution is warranted when foreign transgenes are used as reporters in gene therapy studies with vectors with broader tropism than AAV2. PMID- 19292609 TI - Academy of Breastfeeding Medicine's primary goal is to educate physicians worldwide in breastfeeding and human lactation. PMID- 19292608 TI - Academy of Breastfeeding Medicine founder's lecture 2008: breastfeeding--an extrauterine link between mother and child. AB - In addition to a near-optimal combination of nutrients for the growing infant, breastmilk contains a wide array of bioactive molecules that are known to protect the infant against infectious disease and modulate the composition of the indigenous intestinal microbiota. A growing number of factors that modulate the infant's immunophysiology have also been identified in breastmilk. We suggest that this early immunomodulation via breastmilk is vital for infant health and may explain the epidemiological data indicating that breastmilk reduces the risk of immunoinflammatory conditions in infancy and also later in life. The body of scientific data regarding the role of transforming growth factor-beta in breastmilk in enhancing healthy immune maturation and reducing the risk of disease is reviewed in this article. PMID- 19292611 TI - La Leche League International (LLLI) is foundational to all aspects of breastfeeding. PMID- 19292614 TI - Diphenylhydantoin inhibits osteoclast differentiation and function through suppression of NFATc1 signaling. AB - Diphenylhydantoin (DPH) is widely used as an anticonvulsant drug. We examined the effects of DPH on osteoclast differentiation and function using in vivo and in vitro assay systems. Transgenic mice overexpressing a soluble form of RANKL (RANKL Tg) exhibited increased osteoclastic bone resorption. Injection of DPH into the subcutaneous tissue overlying calvaria of RANKL Tg mice suppressed the enhanced resorption in the calvaria. In co-cultures of mouse osteoblasts and bone marrow cells, DPH inhibited lipopolysaccharide (LPS)-induced osteoclast formation. DPH affected neither the mRNA expression of RANKL and osteoprotegerin nor the growth of mouse osteoblasts in culture. On the other hand, DPH inhibited the RANKL-induced formation of osteoclasts in cultures of mouse bone marrow derived macrophages (BMMphis) and of human peripheral blood-derived CD14(+) cells. DPH concealed LPS-induced bone resorption in mouse calvarial organ cultures and inhibited the pit-forming activity of mouse osteoclasts cultured on dentine slices. DPH suppressed the RANKL-induced calcium oscillation and expression of nuclear factor of activated T cells c1 (NFATc1) and c-fos in BMMphis. Moreover, DPH inhibited the RANKL-induced nuclear localization and auto amplification of NFATc1 in mature osteoclasts. Both BMMphis and osteoclasts expressed mRNA of a T-type calcium channel, Cav3.2, a target of DPH. Blocking the expression of Cav3.2 by short hairpin RNAs significantly suppressed RANKL-induced osteoclast differentiation. These results suggest that DPH inhibits osteoclast differentiation and function through suppression of NFATc1 signaling. The topical application of DPH may be a therapeutic treatment to prevent bone loss induced by local inflammation such as periodontitis. PMID- 19292615 TI - Pro-inflammatory cytokines TNF-related weak inducer of apoptosis (TWEAK) and TNFalpha induce the mitogen-activated protein kinase (MAPK)-dependent expression of sclerostin in human osteoblasts. AB - We have recently shown that TNF-related weak inducer of apoptosis (TWEAK) is a mediator of inflammatory bone remodeling. The aim of this study was to investigate the role of TWEAK in modulating human osteoblast activity, and how TWEAK and TNFalpha might interact in this context. Recombinant TWEAK and TNF were both mitogenic for human primary osteoblasts (NHBC). TWEAK dose- and time dependently regulated the expression of the osteoblast transcription factors RUNX2 and osterix. TWEAK inhibited in vitro mineralization and downregulated the expression of osteogenesis-associated genes. Significantly, TWEAK and TWEAK/TNF induced the expression of the osteoblast differentiation inhibitor and SOST gene product, sclerostin. Sclerostin induction was mitogen-activated protein kinase (MAPK) dependent. The SOST mRNA levels induced by TWEAK were equivalent to or exceeded those seen in steady-state human bone, and the TWEAK/TNF induction of SOST mRNA was recapitulated in fresh cancellous bone explants. TWEAK-induced sclerostin expression was observed in immature osteoblastic cells, both in cycling (Ki67(+)) primary NHBC and in the cell lines MC3T3-E1 and MG-63, as well as in human osteocyte-like cells and in the osteocyte cell line, MLO-Y4. Treatment of NHBC with recombinant human sclerostin mimicked the effects of TWEAK to suppress RUNX2 and osteocalcin (OCN). TWEAK, TNF, and sclerostin treatment of NHBC similarly altered levels of phosphorylated and total GSK3beta and active and total levels of beta-catenin, implying that the Wnt signaling pathway was affected by all three stimuli. Sclerostin also rapidly activated ERK-1/2 MAPK signaling, indicating the involvement of additional signaling pathways. Together, our findings suggest that TWEAK, alone and with TNF, can regulate osteoblast function, at least in part by inducing sclerostin expression. Our results also suggest new roles and modes of action for sclerostin. PMID- 19292616 TI - 1alpha,25(OH)(2) vitamin D(3) induction of ATP secretion in osteoblasts. AB - In the absence of mechanical stimulation, brief exposure of osteoblasts to 1alpha,25(OH)(2)vitamin D(3) (1,25D) triggers plasma membrane electrical responses that couple to exocytosis. Here we describe for the first time 1,25D induction of exocytotic ATP release in static ROS 17/2.8 and SAOS-2 cells and primary calvarial osteoblasts expressing a vitamin D receptor (VDR). We found that 10 nM 1,25D optimally induced 45 +/- 1% and 40 +/- 1% of partial and complete exocytotic events, respectively, from a 1,25D-sensitive pool of ATP containing secretory vesicles within 60 s. We measured a dose-dependent 1,25D induction of ATP secretion, with maximal response of approximately 6.2-fold (16.93 +/- 1.82 nM for SAOS-2) and 3.1-fold (18.89 +/- 1.39 nM for ROS 17/2.8) obtained with 10 nM 1,25D compared with basal ATP levels (2.75 +/- 0.39 nM, SAOS 2; 6.09 +/- 0.58 nM, ROS 17/2.8 cells). The natural metabolite 25(OH)vitamin D(3) (25D, 10 nM) induced a significant 3.6-fold increase of ATP release in ROS 17/2.8 cells, but there was no induction with the antagonist 1beta,25(OH)(2)vitamin D(3) (1beta,25D, 10 nM) or the steroid 17beta-estradiol (10 nM). 1,25D-induced ATP secretion was abolished when cells were preincubated with inhibitors of vesicular exocytosis. siRNA VDR silencing prevented 1,25D stimulation of ATP exocytosis in ROS 17/2.8 and SAOS-2 cells. Similarly, 1,25D failed to stimulate ATP exocytosis in primary osteoblasts from a VDR knockout mouse. ATP secretion coupled to 1,25D induction of cytosolic calcium and chloride channel potentiation. Rapid 1,25D stimulation of ATP secretion involving nontranscriptional VDR functions in osteoblasts may help explain 1,25D bone anabolic properties. PMID- 19292618 TI - Ethics in telemedicine research. PMID- 19292617 TI - Does obesity really make the femur stronger? BMD, geometry, and fracture incidence in the women's health initiative-observational study. AB - Heavier individuals have higher hip BMD and more robust femur geometry, but it is unclear whether values vary in proportion with body weight in obesity. We studied the variation of hip BMD and geometry across categories of body mass index (BMI) in a subset of postmenopausal non-Hispanic whites (NHWs) from the Women's Health Initiative Observational Cohort (WHI-OS). The implications on fracture incidence were studied among NHWs in the entire WHI-OS. Baseline DXA scans of hip and total body from 4642 NHW women were divided into BMI (kg/m(2)) categories: underweight (<18.5), healthy weight (18.5-24.9), overweight (25-29.9), and mild (30-34.9), moderate (35-39.9), and extreme obesity (>40). Femur BMD and indices of bone axial (cross-sectional area [CSA]) and bending strength (section modulus [SM]) were extracted from DXA scans using the hip structure analysis (HSA) method and compared among BMI categories after adjustment for height, age, hormone use, diabetes, activity level, femur neck-shaft angle, and neck length. The association between BMI and incident fracture was studied in 78,013 NHWs from the entire WHI-OS over 8.5 +/- 2.6 (SD) yr of follow-up. Fracture incidence (cases/1000 person-years) was compared among BMI categories for hip alone, central body (hip, pelvis, spine, ribs, and shoulder girdle), upper extremity (humerus and distal), and lower extremity (femur shaft and distal but not hip). Femur BMD, CSA, and SM were larger in women with higher BMI, but values scaled in proportion to lean and not to fat or total body mass. Women with highest BMI reported more falls in the 12 mo before enrollment, more prevalent fractures, and had lower measures of physical activity and function. Incidence of hip fractures and all central body fractures declined with BMI. Lower extremity fractures distal to the hip trended upward, and upper extremity incidence was independent of BMI. BMD, CSA, and SM vary in proportion to total body lean mass, supporting the view that bones adapt to prevalent muscle loads. Because lean mass is a progressively smaller fraction of total mass in obesity, femur BMD, CSA, and SM decline relative to body weight in higher BMI categories. Traumatic forces increase with body weight, but fracture rates at the hip and central body were less frequent with increasing BMI, possibly because of greater soft tissue padding. There was no evident protective effect in fracture rates at less padded distal extremity sites. Upper extremity fractures showed no variation with BMI, and lower extremity fracture rates were higher only in the overweight (BMI = 25 29.9 kg/m(2)). PMID- 19292620 TI - WiMAX: will fourth-generation broadband give telemedicine a boost? PMID- 19292621 TI - Still standing: telemedicine devices and fall prevention. PMID- 19292622 TI - Remote-controlled robotic arm for real-time echocardiography: the diagnostic future for patients in rural areas? AB - There exists a great clinical need for improving specialist consultation and utilization of echocardiography in areas remote from hospital-based care. This paper presents the development and first technical assessment of a concept of cardiovascular consultation utilizing long distance, real-time echocardiography as a diagnostic tool in rural areas. The development of CARdiological consultation at a DISTance (CARDISTA) was achieved in three stages, comprising tests of different broadband infrastructures, videoconference systems, microphones, cameras, monitors, and loudspeakers. The CARDISTA concept includes a cardiologist and a sonographer, a robotic arm (Medirob), a portable ultrasound machine, and presently available information technology using an advanced broadband backbone. The three stages provided, with some remaining doubts, echocardiographic examination at a distance comparable to hospital-based examinations. A continuous broadband capacity of 20 megabits per second (Mbps) seemed to be a vital component of CARDISTA for achieving the highest-quality imaging. With this broadband capacity, it was possible to achieve a transmission delay below 200 ms. The technical tests of the CARDISTA concept revealed promising results in enabling long distance real-time echocardiography for specialist consultation. CARDISTA is now ready for clinical testing and evaluation in rural areas for patients with heart diseases, especially heart failure. PMID- 19292623 TI - St. Vincent's Home telehealth for congestive heart failure patients. AB - St. Vincent's Homecare implemented a remote monitoring project in which researchers studied whether the telehome health patients exhibit enhanced clinical outcomes and patient perceptions of telehome healthcare. Fifty congestive heart failure patients (n = 50) participated in this program. Data collection included pre and post Outcome and Assessment Information Set items, 12 Item Short-Form Health Survey and Minnesota Living with Heart Failure Questionnaire, and data from patient charts were used to capture demographic information. In addition, interviews were conducted in order to assess overall perceptions and attitudes. Results indicate significant changes occurring among respondents in three important aspects that impact their quality of life, namely, physical, behavioral, and emotional improvements. Specifically, statistical significance was documented at the 0.05 level regarding improvement for home telehealth patients in shortness of breath, management of oral medications, ability to engage in moderate activities, amount of energy, swelling in legs/ankles, need to sit/lie down during day, fatigue, need for hospitalization, side effects from treatment, and worry. Additionally, patients found the service easy to use and perceived the care they received via telehealth to be as good as regular in-person care. PMID- 19292624 TI - A pre-post evaluation of a telehomecare program in oncology and palliative care. AB - The purpose of this study was to evaluate the effects of a telehomecare intervention implemented in an oncology and palliative care unit in Quebec, Canada. As part of the intervention, nurses working within the oncology and palliative care program were given a tablet PC to use during patients' home visits and capture data related to the patient's encounter. Outcomes of interest were nurses' satisfaction with the software application, perceived quality of care, and individual and group productivity. With regard to productivity, we hypothesized that this system use would increase (1) the number of patients treated by all the nurses involved in the program, (2) the average number of home visits made by a nurse per shift, and (3) the percentage of time nurses spent providing direct care to patients. There was no intention to reduce the number of home visits made to individual patients. In order to assess the effects of the intervention, data collection was performed over three periods, representing the pre-implementation phase and two post-implementation phases, over 8 months. Data were collected using semistructured interviews, questionnaire surveys, and available documents. Findings revealed that nurses showed a positive attitude toward the software application and were satisfied with the quality of the information stored in the system, which appeared to be of great value for recalling patient information during a visit. According to the nurses, the clinical information system increased the completeness and quality of their interventions in the patient's home, mostly in terms of patient assessment, treatment, and education. Finally, in terms of productivity, we observed a significant increase in the number of patients treated by all the nurses (p = 0.04), the average number of home visits made by a nurse per shift (p = 0.02), and the percentage of time spent by nurses providing direct care (p = 0.003). In summary, the telehomecare intervention produced several positive effects that were related to nurses' quality of documentation and productivity. PMID- 19292625 TI - Cost minimization analysis of a store-and-forward teledermatology consult system. AB - The aim of this study was to perform a cost minimization analysis of store-and forward teledermatology compared to a conventional dermatology referral process (usual care). In a Department of Defense (DoD) setting, subjects were randomized to either a teledermatology consult or usual care. Accrued healthcare utilization recorded over a 4-month period included clinic visits, teledermatology visits, laboratories, preparations, procedures, radiological tests, and medications. Direct medical care costs were estimated by combining utilization data with Medicare reimbursement rates and wholesale drug prices. The indirect cost of productivity loss for seeking treatment was also included in the analysis using an average labor rate. Total and average costs were compared between groups. Teledermatology patients incurred $103,043 in total direct costs ($294 average), while usual-care patients incurred $98,365 ($283 average). However, teledermatology patients only incurred $16,359 ($47 average) in lost productivity cost while usual-care patients incurred $30,768 ($89 average). In total, teledermatology patients incurred $119,402 ($340 average) and usual-care patients incurred $129,133 ($372 average) in costs. From the economic perspective of the DoD, store-and-forward teledermatology was a cost-saving strategy for delivering dermatology care compared to conventional consultation methods when productivity loss cost is taken into consideration. PMID- 19292626 TI - Performance study of the wearable one-lead wireless electrocardiographic monitoring system. AB - This study attempts to compare and assess the performance of a wearable electrocardiogram (ECG) using a sensing fabric electrode and a Bluetooth network with a conventional ECG. A one-lead ECG examination was performed using Bioshirt and an iWorx 214 while walking or running at 3, 6, and 9 km per hour. A correlation coefficient of a heart rate variability (HRV) between these two devices was higher than 0.96 and power spectral density of HRV measured also showed an excellent agreement. Thus, both of these two ECG devices showed similar detection capability for R peaks. The measured values for wave duration and intervals of both devices concur with each other. The intensity of noise is controversial. The ECG device using a sensing fabric electrode and a wireless network showed an ECG signal detection and transmission capability similar to that of a conventional ECG device. PMID- 19292627 TI - E-health program for patients with chronic disease. AB - The aim of this pilot study was to examine the feasibility and benefits of a computer Internet-based videoconferencing healthcare support program for community-dwelling older adults with chronic disease. Eighteen participants (n = 18) who met inclusion criteria were recruited from a community health clinic, provided informed signed consent, and were assigned in groups of six to an Internetbased support group program. Where needed, participants were supplied equipment (computers, Web cams, audio headsets) and trained to access an easy-to use, password-protected Web site that uses videoconferencing to support group member-facilitator interactive communication. The aim of the intervention was to support group member bonding and sharing of challenges and strategies for managing a chronic disease. Following 10 weekly professionally facilitated sessions, the groups met weekly in a self-help mode for an additional 3 months. Participants were interviewed at 6-month follow-up. An interview guide was used to gain feedback on using the Internet to access a health service and to ask about the benefits of having participated in the videoconferencing support group program. At follow-up, participants responded positively to using technology to communicate with healthcare professionals and other group members. They also valued the information shared regarding self-care, and reported reduction in feelings of loneliness and isolation. Our Internet-based, videoconferencing intervention program can be viewed as a prototype for designing technology platforms for the delivery of professional healthcare services to home-based older adults with chronic disease. PMID- 19292628 TI - Nonintrusive biological signal monitoring in a car to evaluate a driver's stress and health state. AB - Nonintrusive monitoring of a driver's physiological signals was introduced and evaluated in a car as a test of extending the concept of ubiquitous healthcare to vehicles. Electrocardiogram, photoplethysmogram, galvanic skin response, and respiration were measured in the ubiquitous healthcare car (U-car) using nonintrusively installed sensors on the steering wheel, driver's seat, and seat belt. Measured signals were transmitted to the embedded computer via Bluetooth(R) communication and processed. We collected and analyzed physiological signals during driving in order to estimate a driver's stress state while using this system. In order to compare the effect of stress on physical and mental conditions, two categories of stresses were defined. Experimental results show that a driver's physiological signals were measured with acceptable quality for analysis without interrupting driving, and they were changed meaningfully due to elicited stress. This nonintrusive monitoring can be used to evaluate a driver's state of health and stress. PMID- 19292629 TI - The influence of economic performance on telemedicine provision. AB - Based on the experiences of Taiwan, this study aims to examine how country-level economic performance affects the provision of healthcare through telecommunications systems and how these health services influence individual level health status. Regression analysis is undertaken and an ordinary least squares method is applied. A panel dataset consists of a national sample over the period from 1995 to 2004. Empirical evidences show that increases in gross domestic product (GDP) increase the reported demand for telemedicine, but the price (cost) of telemedicine reduces that demand. In contrast, the price of conventional medical care and the indicator of healthcare environment (per capita national health expenditure) have no impact on the demand for telemedicine. The study provides quantitative estimates of the impact of GDP and telemedicine prices on the reported demand for telemedicine. A 10% increase in GDP increases the demand for telemedicine services by 3.9%. A 10% increase in the price of telemedicine reduces the demand for telemedicine by 7.4%. Furthermore, Taiwan's current GDP is used to estimate the unmet demand for telemedicine. The estimated current unmet demand for telemedicine is more than three times that currently being provided. In a well-run tele-healthcare system, the residents' capacities to generate income result in their health status being enhanced. Eventually, the economy will enjoy a higher level of welfare due to the impact of health and the provision of telemedicine. This is possibly a much more important source of economic performance and is derived from a policy aimed at promoting telemedicine. PMID- 19292630 TI - Integrating scientific evidence to support telehomecare development in a remote region. AB - This study aimed to understand how different types of knowledge have influenced the decision making process regarding the implementation of telehomecare in the organization of regional healthcare services in the Province of Quebec (Canada). A case study was conducted in order to explore how scientific evidence was integrated in the decision-making processes regarding the implementation of a telehomecare system in the Gaspesie-Magdalene Islands Health Region. A total of 14 semistructured interviews were completed with key organizational decision makers (regional managers, organization managers, healthcare professionals, and technological managers). Two researchers independently carried out data analysis, encouraging iterations and validation with study participants. The Gaspesie Magdalene Islands Telehomecare Project is based on a technological solution named Intelligent Distance Patient Monitoring and constitutes a relevant example of the evolution of an e-health solution. Indeed, the first reports of the experiment influenced decision makers to continue the deployment of the solution. Decision makers from all groups agreed on the importance of using past experience to avoid pitfalls and ensure an optimal decision-making process. They highlighted the importance of knowledge translation between sites as well as within sites. Knowledge translation played an important part in the success of the project. Efficient strategies to transfer evidence to organizational decision making have been identified such as an endusers forum, where researchers provide support by sharing evidence with end-users and actively participate in knowledge translation. PMID- 19292631 TI - NHS Direct telephone helpline: frequency of use over time and by age and gender in an outpatient population. AB - The frequency of awareness and use of the United Kingdom National Health Service (NHS) Direct telephone helpline was measured by questioning consecutive new outpatient referrals seen in a neurology clinic over 3-month periods (January March) in the years 2002-2008. In a cohort of over 1,500 patients, nearly 60% reported awareness of NHS Direct, with increasing cumulative summed frequency of awareness over time, and nearly 20% had used the service with a similar time trend. Analysis by age showed maximal use in the 31-40-year age group with relative underuse by those older than 50 years. Analysis by gender showed that women were more likely both to have heard of and to have used NHS Direct. These findings indicate a steady growth in awareness and use of NHS Direct, yet large numbers of patients reaching secondary care have not utilized this primary care resource. PMID- 19292632 TI - Comment on Internet use for health-related purposes in Greece. PMID- 19292636 TI - A man presenting with nodules on hands and elbows. PMID- 19292638 TI - Uncertainty as to whether the use of antiretroviral therapy for persons recently infected with HIV has a favorable risk-to-benefit ratio. PMID- 19292640 TI - Difficulties in the diagnosis and treatment of acute schistosomiasis. PMID- 19292642 TI - Broadened definition of health care-associated infective endocarditis and risk of misclassification for some community-acquired episodes. PMID- 19292644 TI - 20 years of HIV-2 infection in Portugal: trends and changes in epidemiology. PMID- 19292645 TI - Generalization of the event-based Carnevale-Hines integration scheme for integrate-and-fire models. AB - An event-based integration scheme for an integrate-and-fire neuron model with exponentially decaying excitatory synaptic currents and double exponential inhibitory synaptic currents has been introduced by Carnevale and Hines. However, the integration scheme imposes nonphysiological constraints on the time constants of the synaptic currents, which hamper its general applicability. This letter addresses this problem in two ways. First, we provide physical arguments demonstrating why these constraints on the time constants can be relaxed. Second, we give a formal proof showing which constraints can be abolished. As part of our formal proof, we introduce the generalized Carnevale-Hines lemma, a new tool for comparing double exponentials as they naturally occur in many cascaded decay systems, including receptor-neurotransmitter dissociation followed by channel closing. Through repeated application of the generalized lemma, we lift most of the original constraints on the time constants. Thus, we show that the Carnevale Hines integration scheme for the integrate-and-fire model can be employed for simulating a much wider range of neuron and synapse types than was previously thought. PMID- 19292646 TI - A multiclass classification method based on decoding of binary classifiers. AB - In this letter, we present new methods of multiclass classification that combine multiple binary classifiers. Misclassification of each binary classifier is formulated as a bit inversion error with probabilistic models by making an analogy to the context of information transmission theory. Dependence between binary classifiers is incorporated into our model, which makes a decoder a type of Boltzmann machine. We performed experimental studies using a synthetic data set, data sets from the UCI repository, and bioinformatics data sets, and the results show that the proposed methods are superior to the existing multiclass classification methods. PMID- 19292647 TI - Maximally reliable Markov chains under energy constraints. AB - Signal-to-noise ratios in physical systems can be significantly degraded if the outputs of the systems are highly variable. Biological processes for which highly stereotyped signal generations are necessary features appear to have reduced their signal variabilities by employing multiple processing steps. To better understand why this multistep cascade structure might be desirable, we prove that the reliability of a signal generated by a multistate system with no memory (i.e., a Markov chain) is maximal if and only if the system topology is such that the process steps irreversibly through each state, with transition rates chosen such that an equal fraction of the total signal is generated in each state. Furthermore, our result indicates that by increasing the number of states, it is possible to arbitrarily increase the reliability of the system. In a physical system, however, an energy cost is associated with maintaining irreversible transitions, and this cost increases with the number of such transitions (i.e., the number of states). Thus, an infinite-length chain, which would be perfectly reliable, is infeasible. To model the effects of energy demands on the maximally reliable solution, we numerically optimize the topology under two distinct energy functions that penalize either irreversible transitions or incommunicability between states, respectively. In both cases, the solutions are essentially irreversible linear chains, but with upper bounds on the number of states set by the amount of available energy. We therefore conclude that a physical system for which signal reliability is important should employ a linear architecture, with the number of states (and thus the reliability) determined by the intrinsic energy constraints of the system. PMID- 19292648 TI - Validation-based sparse Gaussian process classifier design. AB - Gaussian processes (GPs) are promising Bayesian methods for classification and regression problems. Design of a GP classifier and making predictions using it is, however, computationally demanding, especially when the training set size is large. Sparse GP classifiers are known to overcome this limitation. In this letter, we propose and study a validation-based method for sparse GP classifier design. The proposed method uses a negative log predictive (NLP) loss measure, which is easy to compute for GP models. We use this measure for both basis vector selection and hyperparameter adaptation. The experimental results on several real world benchmark data sets show better or comparable generalization performance over existing methods. PMID- 19292649 TI - Combining feature- and correspondence-based methods for visual object recognition. AB - We present an object recognition system built on a combination of feature- and correspondence-based pattern recognizers. The feature-based part, called preselection network, is a single-layer feedforward network weighted with the amount of information contributed by each feature to the decision at hand. For processing arbitrary objects, we employ small, regular graphs whose nodes are attributed with Gabor amplitudes, termed parquet graphs. The preselection network can quickly rule out most irrelevant matches and leaves only the ambiguous cases, so-called model candidates, to be verified by a rudimentary version of elastic graph matching, a standard correspondence-based technique for face and object recognition. According to the model, graphs are constructed that describe the object in the input image well. We report the results of experiments on standard databases for object recognition. The method achieved high recognition rates on identity and pose. Unlike many other models, it can also cope with varying background, multiple objects, and partial occlusion. PMID- 19292650 TI - Effect of fiber diameter and alignment of electrospun polyurethane meshes on mesenchymal progenitor cells. AB - Effective strategies to guide cell alignment and the deposition of an oriented extracellular matrix are critical for the development of anisotropic engineered tissues suitable for the repair of ligament defects. Electrospinning is a promising means to create meshes that can align adherent cells, but the effect of fiber mesh architecture on differentiation has not been examined closely. Therefore, the goal of this study was to determine the effect of fiber diameter and the degree of fiber alignment on mesenchymal progenitor cell morphology, proliferation, and ligament gene expression. Specifically, a poly(ester urethane)urea elastomer was electrospun onto rigid supports under conditions designed to independently vary the mean fiber diameter (from 0.28 to 2.3 microm) and the degree of fiber alignment. Bone marrow stromal cells--seeded onto supported meshes--adhered to and proliferated on all surfaces. Cells assumed a more spindle-shaped morphology with increasing fiber diameter and degree of fiber alignment, and oriented parallel to fibers on aligned meshes. Expression of the ligament markers collagen 1alpha1, decorin, and tenomodulin appeared to be sensitive to fiber diameter and greatest on the smallest fibers. Concurrently, expression of the transcription factor scleraxis appeared to decrease with increasing fiber alignment. These results suggest that the formation of a ligament-like tissue on electrospun scaffolds is enhanced when the scaffolds consist of aligned submicron fibers. PMID- 19292651 TI - Moffet on the similarity of response to "active" and "sham" acupuncture. PMID- 19292652 TI - Attitudes of patients with diabetes about complementary medicine in Israel: a cross-cultural perspective. PMID- 19292653 TI - Effects of auricular acupressure on menstrual symptoms and nitric oxide for women with primary dysmenorrhea. AB - OBJECTIVES: The aim of this study was to evaluate the effects of auricular acupressure on relieving menstrual symptoms and decreasing nitric oxide (NO) for women with primary dysmenorrhea. DESIGN: This was a randomized clinical trial comparing the effects of auricular acupressure by seed-pressure method and placebo adhesive patch. SETTING: Settings included colleges in northern and central Taiwan. SUBJECTS: Serum CA-125 testing was used as a screening test for primary dysmenorrhea (<35 mg/dL). The study included 36 college females randomized to acupressure group, 35 to control group. INTERVENTIONS: The acupressure group received auricular acupressure by seed-pressure method on liver (CO12), kidney (CO10), and endocrine (CO18) acupoints. The control group had a plain adhesive patch placed on the same acupoints with no seed attached. Acupressure protocol included massaging 15 times on each acupoint, 3 times a day, for a total of 20 days. OUTCOME MEASURES: Primary: Short-form Menstrual Distress Questionnaire (MDQs). Secondary: blood sample of NO. Assessments of MDQs and NO were performed at baseline and within the first 2 days of their next menses (after completion of 20 days of acupressure). RESULTS: In the acupressure group, the overall menstrual symptoms (95% confidence interval [CI] = -49.8 to -6.5, effect size [ES] = 0.43, p = 0.01) and two subscales, menstrual pain (95% CI = 16.4 -to -2.2, ES = 0.45, p = 0.01) and negative affects (95% CI = -11.9-2.0, ES = 0.38, p = 0.04), revealed that menstrual symptoms decreased significantly after auricular acupressure by the seed-pressure method. The ES for the MDQs were in favor of the auricular acupressure by seed-pressure method. NO level increased in the acupressure group, although this difference did not achieve statistical significance (p > 0.05). CONCLUSIONS: This study supports the effects of auricular acupressure by seed-pressure method in improving menstrual symptoms, and offers a noninvasive complementary therapy for women with primary dysmenorrhea. PMID- 19292654 TI - Antioxidative potentiality of a Kampo formulation measured by an ex vivo study. AB - OBJECTIVES: Antioxidants in herbs or crude herbal formulations are well known. The antioxidative effect of a Kampo formulation, Bofu-tsusho-san (BTS), was studied in healthy human subjects by means of an ex vivo study of the lag time of low-density lipoprotein (LDL) oxidation. METHODS: Eighteen (18) healthy young males (22 +/- 3 years old), who were screened out from 38 males by strict screening tests of serum lipids, were randomly assigned into 3 groups (active BTS 7.5 g/day, placebo of BTS 7.5 g/day, and vitamin E 500 mg and vitamin C 1000 mg/day). The BTS groups were assigned in a double-blind manner. These 3 groups took each test drug, and the antioxidant effects of their plasma were estimated by assessing the susceptibility of LDL to oxidation induced by 2-2'-azobis (4 methoxy-2,4-dimethyl-valeronitrile) via an ex vivo study. The antioxidative activity was evaluated by measuring the lag time of the production of conjugated dienes. Along with lag time, plasma ephedrine, plasma baicalin, serum lipid peroxide, serum free fatty acids (nonesterified fatty acids), urinary 8(OH)dG/creatinine, blood pressure, and heart rate were monitored for 24 hours. RESULTS: Although lag time of the placebo group revealed no significant changes, the active BTS group showed some tendency of prolonged plasma lag time (p = 0.08). However, urinary 8(OH)dG/creatinine and serum lipid peroxide did not show no significant changes. The physiologic changes were compatible with the pharmacokinetic results of ephedrine and its sympathomimetic effect in the active BTS group. CONCLUSIONS: Although the systemic antioxidative effects of BTS have not been ascertained, the antioxidative potentiality on LDL oxidation was suggested by the ex vivo study. PMID- 19292655 TI - The development of a prospective data collection process in a traditional Chinese medicine teaching clinic. AB - OBJECTIVE: There is a growing need for students and practitioners of Traditional Chinese Medicine to gain experience with standardized data collection, patient outcomes measurement, and practice-based research. The purpose of this paper is to describe the development of a process for standardized data collection that could eventually be adopted for clinical, research, and quality assurance purposes. SETTINGS/LOCATION: The setting for this study was an acupuncture and Oriental medicine teaching clinic in Bloomington, Minnesota. METHODS: Four (4) aspects of data collection were assessed and improved, including intake and post treatment questionnaires, follow-up with patients, integration of data collection into clinic flow, and commitment of resources to the project. OUTCOME MEASURES: The outcomes measures were data collection and missing data rates, burden on patients and clinic staff, and efficiency of data entry. RESULTS: Revision to the data collection process resulted in decreased burden to patients and staff, more detailed and aggressive follow-up protocols, enhanced training for clinic staff, and increased personnel and data-related resources. CONCLUSIONS: The systematic collection of descriptive and clinical characteristics can be accomplished in a teaching clinic with thoughtful attention paid to data collection protocols, dedicated resources, and the involvement of all relevant personnel. PMID- 19292656 TI - Cerebral hemodynamic predictors of poor 6-month Glasgow Outcome Score in severe pediatric traumatic brain injury. AB - Little is known regarding the cerebral autoregulation in pediatric traumatic brain injury (TBI). We examined the relationship between cerebral hemodynamic predictors, including cerebral autoregulation, and long-term outcome after severe pediatric TBI. After Institutional Review Board (IRB) approval, a retrospective analysis of prospectively collected data (May 2002 to October 2007) for children age < or =16 years with severe TBI (admission Glasgow Coma Scale [GCS] score <9) was performed. Cerebral autoregulation was assessed within 72 h after TBI. Cerebral hemodynamic predictors (intracranial pressure [ICP], systolic blood pressure [SBP], and cerebral perfusion pressure [CPP]) through the first 72 h after TBI were abstracted. Univariate and multivariate analyses examined the relationship between impaired cerebral autoregulation (autoregulatory index <0.4), intracranial hypertension (ICP >20 mm Hg), and hypotension (SBP <5th percentile and CPP <40 mm Hg). Six-month Glasgow Outcome Scale (GOS) score of <4 defined poor outcome. Ten (28%) of the 36 children examined (9.1 +/- 5.3 [0.8-16] years; 74% male) had poor outcome. Univariate factors associated with poor outcome were impaired cerebral autoregulation (p = 0.005), SBP <5(th) percentile for age and gender (p = 0.02), and low middle cerebral artery flow velocity (<2 SD for age and gender; p = 0.04). Independent risk factors for poor 6-month GOS were impaired cerebral autoregulation (adjusted odds ratio [aOR] 12.0; 95% confidence interval [CI] 1.4-99.4) and hypotension (SBP <5th percentile; aOR 8.8; 95% CI 1.1-70.5), respectively. Previous studies of TBI describing poor outcome with hemodynamics did not consider the status of cerebral autoregulation. In this study, both impaired cerebral autoregulation and SBP <5th percentile were independent risk factors for poor 6-month GOS. PMID- 19292657 TI - Effects of white, grey, and pia mater properties on tissue level stresses and strains in the compressed spinal cord. AB - Recent demographics demonstrate an increase in the number of elderly spinal cord injury patients, motivating the desire for a better understanding of age effects on injury susceptibility. Knowing that age and disease affect neurological tissue, there is a need to better understand the sensitivity of spinal cord injury mechanics to variations in tissue behavior. To address this issue, a plane strain, geometrically nonlinear, finite element model of a section of a generic human thoracic spinal cord was constructed to model the response to dorsal compression. The material models and stiffness responses for the grey and white matter and pia mater were varied across a range of reported values to observe the sensitivity of model outcomes to the assigned properties. Outcome measures were evaluated for percent change in magnitude and alterations in spatial distribution. In general, principal stresses (114-244% change) and pressure (75 119% change) were the outcomes most sensitive to material variation. Strain outcome measures were less sensitive (7-27% change) than stresses (74-244% change) to variations in material tangent modulus. The pia mater characteristics had limited (<4% change) effects on outcomes. Using linear elastic models to represent non-linear behavior had variable effects on outcome measures, and resulted in highly concentrated areas of elevated stresses and strains. Pressure measurements in both the grey and white matter were particularly sensitive to white matter properties, suggesting that degenerative changes in white matter may influence perfusion in a compressed spinal cord. Our results suggest that the mechanics of spinal cord compression are likely to be affected by changes in tissue resulting from aging and disease, indicating a need to study the biomechanical aspects of spinal cord injury in these specific populations. PMID- 19292658 TI - Development of three-dimensional tissue-engineered models of bacterial infected human skin wounds. AB - While infected skin wounds are on the increase because of ageing populations, rising incidence of diabetes, and antibiotic resistance, we lack relevant in vivo or in vitro models to study many aspects of bacterial interaction with skin. The aim of this study was to develop three-dimensional models of normal human skin to study bacterial infection. The common dermatological pathogens Staphylococcus aureus and Pseudomonas aeruginosa were used to infect tissue-engineered skin, and the course of infection in the skin was examined over several days. Two forms of model were developed-one in which bacteria were introduced directly to 10 mm wounds in the epidermis, and another in which wounds were created by burning a 4 mm hole in the center of the tissue before inoculation. The bacteria flourished within the engineered skin, and colonized the upper epidermal layers before invasion into the dermis. Infection with P. aeruginosa caused a loss of epidermis and de-keratinization of the skin constructs, as well as partial loss of basement membrane. These novel complex human skin infection models could be used to investigate microbial invasion of normal skin epithelium, basement membrane, and connective tissue, and as a model to study approaches to reduce bacterial burden in skin wounds. PMID- 19292659 TI - The effect of hypoxia on in vitro prevascularization of a thick soft tissue. AB - Prevascularizing an implantable tissue is one strategy to improve oxygen (O(2)) transport throughout larger tissues upon implantation. This study examined the role of hypoxia both during (i.e., as a stimulus) and after (i.e., mimicking implant conditions) vascularization of an implantable tissue. Tissues consisted of microcarrier beads coated with human umbilical vein endothelial cells embedded in fibrin. The fibrin was covered with a monolayer of normal human lung fibroblasts (NHLFs), or exposed to conditioned media from NHLFs. Capillary networks developed at 20% or 1% O(2) tension for 8 days. In some experiments, tissues were supplemented with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor, whereas in others the tissues prevascularized at 20% O(2) were transferred to 1% O(2) for 8 additional days. Maximal capillary formation occurred in media conditioned by NHLFs at 20% O(2), supplemented with VEGF (concentration >10 pM). Hypoxia (1% O(2)) did not stimulate basic fibroblast growth factor production and decreased in vitro angiogenesis, despite an increase in endogenous VEGF production. Hypoxia also degraded a preformed capillary network within 4 days. Hence, strategies to prevascularize implantable tissues may not require the physical presence of stromal cells, but will likely require fibroblast-derived growth factors in addition to VEGF to maintain capillary growth. PMID- 19292661 TI - Decreased viral load and symptoms of polyomavirus-associated chronic interstitial cystitis after intravesical cidofovir treatment. AB - Interstitial cystitis causes disabling bladder pain and is usually diagnosed in the absence of infection. We describe a patient with interstitial cystitis who had high urinary levels of polyomavirus that decreased dramatically after initiation of intravesical cidofovir treatment; the patient also showed substantial improvement in symptoms. Another patient had milder symptoms of cystitis and intermittent polyomavirus shedding. Polyomaviruses, particularly BK virus, may cause some cases of interstitial cystitis. PMID- 19292660 TI - Risk factors for fecal colonization with multiple distinct strains of Escherichia coli among long-term care facility residents. AB - Of 49 long-term care facility residents, 21 (43%) were colonized with 2 or more distinct strains of Escherichia coli. There were no significant risk factors for colonization with multiple strains of E. coli. These results suggest that future efforts to efficiently identify the diversity of colonizing strains will be challenging. PMID- 19292662 TI - Ethical considerations about reporting research results with potential for further stigmatization of undocumented immigrants. AB - A broad spectrum of infectious diseases is studied in vulnerable populations. However, ethical considerations of reporting research results that could increase stigmatization of socially marginalized and vulnerable populations are not often discussed in the medical literature, particularly not in the context of transmissible diseases. This article addresses ethical considerations that arose when one of us (J.M.A.) recently published the results of a study in Clinical Infectious Diseases that imply that undocumented persons are more likely to transmit tuberculosis than are documented foreign-born persons or persons born in the United States. These study results have the potential to further fuel the often fierce debate regarding undocumented immigrants in the United States. To our knowledge, such ethical considerations have not been discussed previously in the medical literature. PMID- 19292663 TI - Catheter-associated urinary tract infections: a syllogism compounded by a questionable dichotomy. PMID- 19292664 TI - Inappropriate treatment of catheter-associated asymptomatic bacteriuria in a tertiary care hospital. AB - BACKGROUND: Evidence-based guidelines state that asymptomatic bacteriuria is not a clinically significant condition in men and nonpregnant women and that treatment is unlikely to confer clinical benefit. We hypothesized that, among patients with indwelling catheters or condom collection systems, many who receive a diagnosis of and are treated for catheter-associated urinary tract infection (CAUTI) actually have asymptomatic bacteriuria and, therefore, that antibiotic therapy is inappropriate. METHODS: We reviewed all urine culture results at a veterans affairs medical center during a 3-month period. Cultures yielding 10(4) colony-forming units/mL were included if the urine had been collected from a hospitalized patient with an indwelling (Foley) catheter or a condom collection system. We applied standardized definitions to determine whether the episode represented catheter-associated asymptomatic bacteriuria (CAABU) or CAUTI. Antibiotic therapy was considered appropriate for patients who met criteria for symptomatic UTI. RESULTS: Overall, 280 episodes met criteria for inclusion: 164 CAABU and 116 CAUTI. Of the 164 episodes of CAABU, 111 (68%) were managed appropriately (no treatment), whereas 53 (32%) were treated with antibiotics (inappropriate treatment). In multivariate analysis, older patient age, having predominantly gram-negative bacteriuria, and higher urine white blood cell count were significantly associated with inappropriate treatment of CAABU (P < .05, by logistic regression). CONCLUSIONS: Better recognition of CAABU and the distinction between this condition and CAUTI, consistent with evidence-based guidelines, may play a key role in reducing unneeded antibiotic usage in hospitalized patients. PMID- 19292665 TI - Lack of efficacy of prophylactic platelet transfusion for severe thrombocytopenia in adults with acute uncomplicated dengue infection. AB - Thrombocytopenia in dengue infection raises concerns about bleeding risk. Of 256 patients with dengue infection who developed thrombocytopenia (platelet count, < 20 x 10(3) platelets/microL) without prior bleeding, 188 were given platelet transfusion. Subsequent bleeding, platelet increment, and platelet recovery were similar between patients given transfusion and patients not given transfusion. Prophylactic platelet transfusion was ineffective in preventing bleeding in adult patients with dengue infection. PMID- 19292666 TI - Muscle differentiation and myotubes alignment is influenced by micropatterned surfaces and exogenous electrical stimulation. AB - An in vitro muscle-like structure with parallel-oriented contractile myotubes is needed as a model of muscle tissue regeneration. For this purpose, it is necessary to reproduce a controllable microscale environment mimicking the in vivo cues. In this work we focused on the application of topological and electrical stimuli on muscle precursor cell (MPC) culture to influence MPC orientation and induce myotube alignment. The two stimulations were tested both independently and together. A structural and topological template was achieved using micropatterned poly-(L-lactic acid) membranes. Electrical stimulation, consisting of square pulses of 70 mV/cm amplitude each 30 s, was applied to the MPC culture. The effect of different pulse durations on cultures was evaluated by galvanotaxis analysis. The highest cell displacement rate toward the cathode was observed for 3 ms pulse stimulation, which was then applied in combination with topological stimuli. Topological and electrical stimuli had an additive effect in enhancing differentiation of cultured MPC, shown by high Troponin I protein production and, in parallel, Myogenin and Desmin genes, down- and upregulation respectively. PMID- 19292667 TI - Construction of synthetic dermis and skin based on a self-assembled peptide hydrogel scaffold. AB - Using biocompatible peptide hydrogel as a scaffold, we prepared three-dimensional synthetic skin that does not contain animal-derived materials or pathogens. The present study investigated preparation methods, proliferation, and functional expression of fibroblasts in the synthetic dermis and differentiation of keratinocytes in the epidermis. Synthetic dermis was prepared by mixing fibroblasts with peptide hydrogel, and synthetic skin was prepared by forming an epidermal layer using keratinocytes on the synthetic dermis. A fibroblast-rich foamy layer consisting of homogeneous peptide hydrogel subsequently formed in the synthetic dermis, with fibroblasts aggregating in clusters within the septum. The epidermis consisted of three to five keratinocyte layers. Immunohistochemical staining showed human type I collagen, indicating functional expression around fibroblasts in the synthetic dermis, keratinocyte differentiation in the epidermis, and expression of basement membrane proteins. The number of fibroblasts tended to increase until the second week and was maintained until the fourth week, but rapidly decreased in the fifth week. In the synthetic dermis medium, the human type I collagen concentration increased after the second week to the fifth week. These findings suggest that peptide hydrogel acts as a synthetic skin scaffold that offers a platform for the proliferation and functional expression of fibroblasts and keratinocytes. PMID- 19292668 TI - Physicobiochemical synergism through gene therapy and functional tissue engineering for in vitro chondrogenesis. AB - Mechanical and chemical stimulation have been shown to enhance in vitro chondrogenesis. The aim of this study was to analyze and compare combined physicobiochemical effects. Bovine articular chondrocytes were retrovirally transduced to express bone morphogenetic protein-2 (BMP-2) or left as naive controls. Cells were seeded in three-dimensional polyurethane scaffolds and further cultured under static conditions or exposed to dynamic compression and shear in a joint-specific bioreactor. Four groups: control (A), load (B), BMP-2 infected (C), and BMP-2-infected plus load (D) were analyzed for DNA and glycosaminoglycan (GAG) content; collagen I, II, and X; aggrecan, (cartilage oligomeric protein (COMP), superficial zone protein, matrix metalloproteinase (MMP)-3; MMP-13 mRNA; histology; and immunohistochemistry at 7, 21, and 35 days post-seeding. Synergistic effects (D) were higher than the sum of the individual treatments (B and C) for GAG/DNA, collagen II, and COMP. Histology revealed a functional organization in D including an intense safranin O staining in C and D superior to that in A and B. Immunostaining for collagen II and aggrecan was detected in C and D and was strongest in D. The results show that both stimuli augment in vitro chondrogenesis better than in controls. Biochemical manipulation proved to be predominantly more effective than load, and synergistic effects were demonstrated. PMID- 19292669 TI - Novel effect of biphasic electric current on in vitro osteogenesis and cytokine production in human mesenchymal stromal cells. AB - Electrical stimulation (ES) can activate diverse biostimulatory responses in a range of tissues. Of various forms of ES, the application of biphasic electric current (BEC) is a new approach to bone formation. This study is to investigate the effects and mechanism of action of BEC in osteoblast differentiation and cytokine production in human mesenchymal stromal cells (hMSCs). Using an in vitro culture system with a modified version of the BEC stimulator chip used in our previous study, we exposed hMSCs to a 100 Hz ES with a magnitude of 1.5/15 muA/cm(2) for 250/25 mus. hMSCs showed increased proliferation during static BEC stimulation for 5 days. However, alkaline phosphatase activity and calcium deposition were enhanced in hMSCs 7 days after the stimulation, rather than during the period of ES. BEC induced vascular endothelial growth factor (VEGF) and BMP-2 production; the former can enhance the proliferation of human umbilical vein endothelial cells in culture using conditioned media from BEC cultures. Treatment with selective inhibitors of p38 MAPK (SB203580) or Erk (PD98059), as well as calcium channel blockers (verapamil and nifedipine), reduced the BEC mediated increase of VEGF expression and cell proliferation. These findings reveal that BEC is involved in the osteoblast differentiation of hMSCs through enhancement of cell proliferation and modulation of the local endocrine environment through VEGF and BMP-2 induction through the activation of MAPK (Erk and p38) and the calcium channel. Thus, local stimulation using BEC might be most beneficial in promoting osteogenic differentiation of hMSCs, resulting in enhanced bone formation for bone tissue engineering. PMID- 19292670 TI - A student team in a University of Michigan biomedical engineering design course constructs a microfluidic bioreactor for studies of zebrafish development. AB - The zebrafish is a valuable model for teaching developmental, molecular, and cell biology; aquatic sciences; comparative anatomy; physiology; and genetics. Here we demonstrate that zebrafish provide an excellent model system to teach engineering principles. A seven-member undergraduate team in a biomedical engineering class designed, built, and tested a zebrafish microfluidic bioreactor applying microfluidics, an emerging engineering technology, to study zebrafish development. During the semester, students learned engineering and biology experimental design, chip microfabrication, mathematical modeling, zebrafish husbandry, principles of developmental biology, fluid dynamics, microscopy, and basic molecular biology theory and techniques. The team worked to maximize each person's contribution and presented weekly written and oral reports. Two postdoctoral fellows, a graduate student, and three faculty instructors coordinated and directed the team in an optimal blending of engineering, molecular, and developmental biology skill sets. The students presented two posters, including one at the Zebrafish meetings in Madison, Wisconsin (June 2008). PMID- 19292671 TI - Characterization and in vitro control of MPF activity in zebrafish eggs. AB - We describe the characterization of maturation-promoting factor (MPF) in zebrafish eggs and used different defined conditions to maintain its activity in vitro. MPF activity levels are high in freshly ovulated mature eggs and decline rapidly within 5 min after either fertilization or parthenogenetic activation. The MPF activity of eggs matured in vitro declines faster when the eggs are incubated in Hank's culture medium supplemented with 0.5% BSA (H-BSA) than when incubated in Chinook salmon ovarian fluid (CSOF). MPF activity in nonactivated, aged eggs remains high in H-BSA supplemented with 75 microM MG132 or 10 mM caffeine, but neither MG132 nor caffeine can sustain high MPF activity in activated eggs. MG132-treated eggs showed delayed completion of metaphase and extrusion of the second polar body. Nuclear staining of the activated eggs confirmed the correlation between their cell cycle stage and MPF activity at each time point. An embryotoxic effect was found when matured eggs were held in 100 microM of MG132 or 20 mM caffeine for 1 h. Calcium-depleted medium and 1,2-bis(o aminophenoxy)ethane-N,N,N',N'-tetraacetic acid also showed detrimental effects on the embryos. Conversely, nonactivated, aged matured eggs maintained high MPF activity and developmental potential when CSOF was used as a holding medium. PMID- 19292672 TI - Noncanonical Wnt signaling in tumor progression and metastasis. AB - For almost 15 years, the concept that noncanonical (beta-catenin-independent) Wnt signaling pathways play key roles in embryonic development has grown steadily in the scientific literature. Significant progress has been made toward understanding how these pathways regulate morphogenetic processes as diverse as gastrulation cell movements and the formation of cilia. More recently, however, data have implicated components of noncanonical Wnt/Ca(2+) and Wnt/planar cell polarity signaling in directly promoting the invasiveness and malignant progression of diverse forms of human cancer. Here I review this emerging field of cancer research using data from developmental model systems to provide a framework for addressing future questions. PMID- 19292673 TI - Apc1-mediated antagonism of Wnt/beta-catenin signaling is required for retino tectal pathfinding in the zebrafish. AB - The tumor suppressor Apc1 is an intracellular antagonist of the Wnt/beta-catenin pathway. We examined the effects of an Apc1 loss-of-function mutation on retino tectal axon pathfinding in zebrafish. In apc mutants, the retina is disorganized and optic nerves portray pathfinding defects at the optic chiasm and do not project properly to the tectum. Wild-type cells, transplanted into mutant retinae, acquire retinal ganglion cell fate and project axons that cross at the mispositioned optic chiasm and extend to the contralateral tectum, suggesting a function of apc1 in axon pathfinding. These defects are caused mainly by stabilization of beta-catenin. These data demonstrate that Apc1 function is required for correct patterning of the retina and proper retinal ganglion axon projections. PMID- 19292674 TI - Noncanonical Wnt/PCP signaling during vertebrate gastrulation. AB - The branch of the Wnt pathway, related to planar cell polarity signaling in Drosophila, is fundamental not only to the establishment of tissue polarity but also to a variety of morphogenetic processes in vertebrates. The genetic pathway has been noted for its similarity as well as divergence of between vertebrates and Drosophila. This review focuses on issues related to the complexity of the output of the planar cell polarity pathway during gastrulation in zebrafish and Xenopus and, to a lesser extent, during gastrulation/neurulation in mice. PMID- 19292675 TI - Functional tissue engineering requires bioreactor strategies. PMID- 19292676 TI - Osteoblast and osteoclast differentiation in an in vitro three-dimensional model of bone. AB - There is increasing interest in developing new in vitro tissue models using typical tissue engineering approaches. This study was designed to (1) develop a novel three-dimensional (3D) in vitro model of bone by seeding murine primary osteoblasts and osteoclast precursors on a resorbable porous ceramic scaffold based on silicon-stabilized tricalcium phosphate (Skelite), and (2) investigate bone cell interactions in a 3D environment mimicking an in vivo condition and compare it to traditional two-dimensional (2D) cultures. Murine primary osteoblasts from C57Bl6/J mice and osteoclast precursors from C57Bl/6-Tg(ACTB EGFP)1Osb/J mice were co-cultured on 3D Skelite scaffolds and on standard plastic culture dishes. The differentiation of these cells in both culture conditions was compared by histology (hematoxylin-eosin staining and polarized light analysis), immunohistochemistry (collagen type I), and gene expression analysis by real-time PCR for Runt-related transcription factor 2, osterix, osteocalcin, cathepsin K, and tartrate resistant acid phosphatase. To analyze and compare bone turnover in 3D and 2D co-cultures, we evaluated the modulation of RANKL and OPG mRNA expression. We observed an enhancement of osteoblast differentiation in the 3D mineralized environment that in turn promoted earlier osteoclast differentiation. In this paper, we also report that the increased osteoblast differentiation in the 3D model led to a deposition of extracellular matrix that faithfully reflected the morphology of bone tissue. PMID- 19292677 TI - Potential of hydrogels based on poly(ethylene glycol) and sebacic acid as orthopedic tissue engineering scaffolds. AB - In this study, the bioactive effects of poly(ethylene glycol) (PEG) sebacic acid diacrylate (PEGSDA) hydrogels with or without RGD peptide modification on osteogenic differentiation and mineralization of marrow stromal cells (MSCs) were examined. In a separate experiment, the ability of PEGSDA hydrogel to serve as a delivery vehicle for bone morphogenetic protein 2 (BMP-2) was also investigated. As a scaffold, the attachment and proliferation of MSCs on PEGSDA hydrogel scaffolds with and without RGD peptide modification was similar to the control, tissue culture polystyrene. In contrast, cells were barely seen on unmodified PEG diacrylate (PEGDA) hydrogel throughout the culture period for up to 21 days. Osteogenic phenotypic expression such as alkaline phosphatase (ALP) of MSCs as well as mineralized calcium content were significantly higher on PEGSDA-based hydrogels than those on the control or PEGDA hydrogels. Potential use of PEGSDA scaffold as a delivery vehicle of osteogenic molecules such as BMP-2 was also evaluated. Initial burst release of BMP-2 from PEGSDA hydrogel scaffold (14.7%) was significantly reduced compared to PEGDA hydrogel scaffold (84.2%) during the first 3 days of a 21-day release period. ALP activity of an osteoblast was significantly higher in the presence of BMP-2 released from PEGSDA hydrogel scaffolds compared to that in the presence of BMP-2 released from PEGDA scaffolds, especially after 6 days of release. Overall, PEGSDA hydrogel scaffolds without further modification may be useful as orthopedic tissue engineering scaffolds as well as local drug carriers for prolonged sustained release of osteoinductive molecules. PMID- 19292678 TI - Dose-finding study of fibrin gel-immobilized vascular endothelial growth factor 165 and basic fibroblast growth factor in the arteriovenous loop rat model. AB - The angiogenic effects of different concentrations of vascular endothelial growth factor (VEGF) 165 and basic fibroblast growth factor (bFGF) immobilized in a fibrin-based drug-delivery system were quantitatively assessed in the arteriovenous (AV) loop model. An AV loop was created in the medial thigh of 60 rats. The loop was placed in a Teflon isolation chamber and embedded in 500 microL of fibrin gel loaded with VEGF and bFGF in four different concentrations (no growth factor, 100 ng/mL of VEGF, 25 ng/mL of VEGF and bFGF, 100 ng/mL pf VEGF and bFGF). The explantation intervals were 1, 2, and 4 weeks after the initial operation for all groups. Specimens were investigated using (micro-CT) and histological and morphometrical techniques. After 2 weeks, the cross-section area and construct weight were significantly lower with the use of 100 ng/mL of VEGF and bFGF. Micro-CT and histology showed significantly greater vascular density and number of vessels of the constructs at 2 and 4 weeks when 100 ng/mL of VEGF165 and bFGF were applied than in the growth factor-free specimens. The angioinductive effects were dose-dependent, with best results when using 100 ng/mL of VEGF165 and bFGF. The greater tissue formation was accompanied by faster resorption of the fibrin matrix. PMID- 19292679 TI - The effect of perfluorocarbon-based artificial oxygen carriers on tissue engineered trachea. AB - The biological effect of the perfluorocarbon-based artificial oxygen carrier (Oxygent) was investigated in tissue-engineered trachea (TET) construction. Media supplemented with and without 10% Oxygent were compared in all assessments. Partial tissue oxygen tension (PtO(2)) was measured with polarographic microprobes; epithelial metabolism was monitored by microdialysis inside the TET epithelium perfused with the medium underneath. Chondrocyte-DegraPol constructs were cultured for 1 month with the medium before glycosaminoglycan assessment and histology. Tissue reaction of TET epithelial scaffolds immersed with the medium was evaluated on the chick embryo chorioallantoic membrane. Oxygent perfusion medium increased the TET epithelial PtO(2) (51.2 +/- 0.3 mm Hg vs. 33.4 +/- 0.3 mm Hg at 200 microm thickness; 12.5 +/- 0.1 mm Hg vs. 3.1 +/- 0.1 mm Hg at 400 microm thickness, p < 0.01) and decreased the lactate concentration (0.63 +/- 0.08 vs. 0.80 +/- 0.06 mmol/L, p < 0.05), lactate/pyruvate (1.87 +/- 0.26 vs. 3.36 +/- 10.13, p < 0.05), and lactate/glucose ratios (0.10 +/- 0.00 vs. 0.29 +/- 0.14, p < 0.05). Chondrocyte-DegraPol in Oxygent group presented lower glycosaminoglycan value (0.03 +/- 0.00 vs. 0.13 +/- 0.00, p < 0.05); histology slides showed poor acid mucopolysaccharides formation. Orthogonal polarization spectral imaging showed no difference in functional capillary density between the scaffolds cultured on chorioallantoic membranes. The foreign body reaction was similar in both groups. We conclude that Oxygent increases TET epithelial PtO(2), improves epithelial metabolism, does not impair angiogenesis, and tends to slow cartilage tissue formation. PMID- 19292680 TI - Isolation and implantation of bone marrow-derived mesenchymal stem cells with fibrin micro beads to repair a critical-size bone defect in mice. AB - Fibrin microbeads (FMBs) made using thermal treatment of fibrin drops in oil can efficiently isolate mesenchymal stem cells (MSCs) from bone marrow (BM) and other similar sources and culture them continuously in suspension culture. The pure mesenchymal profile of MSCs isolated using FMBs and their differentiation potency to different mesenchymal lineages were previously described in detail. In the current study, MSCs were isolated from the BM of (GFP+) C57/bl mice using FMBs. Addition of pro-osteogenic medium with 10 mM of ss-glycerolphosphate, 50 microg/mL of ascorbic acid, and 10(-8) M of dexamethasone for 1 month resulted in ossified bone-like solid cellular structures, as seen using fluorescence and scanning electron microscopy (SEM). Such spontaneously formed structures were implanted in full-depth approximately 5-mm-diameter drilled defects in the skulls of wild-type c57/bl mice. Two months later, the excised upper parts of the skulls with the defects were viewed using fluorescence microscopy for green fluorescence protein of the cells in the defect and using SEM. They were also scanned using micro-computed tomography to visualize the formation of new hard tissue. Then the samples were processed and sectioned for hematoxylin and eosin staining and immunohistochemistry. Implanted FMBs loaded with (GFP+) MSCs formed partially mature, dense bone-like tissue using a residual moderate inflammatory process containing remnants of FMBs and neo-angiogenesis. The filled defect with bone like tissue had a Ca/P ratio similar to that of native bone. Limited merging of the implant with the skull indicated that the induced bone regeneration derived from the MSCs that were delivered with the implant. No repair was seen in the control animals without implants or where the defect was filled with FMBs only. Repair scoring (on a 0-5 scale) was found to be 3.38+/-0.35 in the experimental arm, relative to 0 in the controls (p < 0.001). PMID- 19292681 TI - Minced skin for tissue engineering of epithelialized subcutaneous tunnels. AB - We used minced, autologous skin for neoepithelialization of surgically created subcutaneous tunnels in a large animal model. Partial-thickness skin grafts were harvested from the back region of five 50-60 kg Yorkshire pigs. The skin was minced to 0.8 x 0.8 x 0.3 mm particles. Silicone-latex tubes were covered with fibrin, rolled in minced skin, and placed in subcutaneous tunnels created in the abdominal area. For comparison, single cell suspensions of keratinocytes and fibroblasts in fibrin or fibrin only were transplanted on tubes. Tunnels were extracted after 14, 21, and 28 days for microscopic evaluation. All tubes transplanted with minced skin particles showed neoepithelialization. The epithelium was stratified and differentiated after 2 weeks in vivo, and the stratum corneum was directed toward the implanted tube. No epithelium formed from tubes transplanted with single cell suspensions, and only sparse keratinocytes could be detected by serial sectioning and immunostaining on day 14, but not later. No epithelial lining was found in tunnels with fibrin-only-coated tubes. Epithelial cysts could be found the first 2 weeks after transplantation in the minced skin group but not later. In conclusion, a minced skin technique could serve as a potential source for tissue engineering of tubular conduits for reconstructive purposes of the urethra and for cutaneous stomas for bladder catheterization, or intestinal irrigations. The method would have the advantage of being simple and expeditious and not requiring in vitro culturing. PMID- 19292682 TI - Human osteoblast-derived factors induce early osteogenic markers in human mesenchymal stem cells. AB - The capacity of human mesenchymal stem cells (hMSC) for self-renewal and differentiation is a tightly regulated process within their microenvironment--the stem cell niche. For future therapeutic applications of hMSC within the frame of tissue engineering, it is of major importance to understand the factors involved in triggering differentiation cascades of hMSC. Using either osteoblast conditioned medium or an indirect coculture system, we investigated whether soluble factors from human osteoblasts (hOB) are sufficient to induce early osteogenic markers in hMSC. Thereby, we detected an induction of several osteogenic markers like alkaline phosphatase, bone sialoprotein 2, leptin receptor, decorin, and cathepsin K in hMSC as indicators of the onset of early osteogenesis. Further, because Wnt signaling has been reported to play an important role in osteogenesis, we performed RNAi against the main Wnt mediator beta-catenin and the low-density lipoprotein receptor-related protein 5 as a major Wnt co-receptor in hMSC. Whereas alkaline phosphatase was significantly downregulated with this approach, the other osteogenic markers showed a markedly upregulation. These observations suggest that hOB-secreted factors could induce early osteogenic markers in hMSC. Thus, with regard to a therapeutic setting, these findings may pave the way for a more in vivo-related differentiation procedure for the generation of osteoblast-like cells. PMID- 19292683 TI - Enhanced ex vivo expansion of human adipose tissue-derived mesenchymal stromal cells by fibroblast growth factor-2 and dexamethasone. AB - In the present study, we investigated the ex vivo expansion of human adipose tissue-derived mesenchymal stromal cells (ATSCs) to identify factors that promoted efficient expansion while preserving stem cell potential. We examined several growth factors and steroids, and found that the combination of a low concentration of fibroblast growth factor-2 (FGF-2) (1 ng/mL) and dexamethasone (DEX) or betamethasone (BET) enhanced the proliferation of ATSCs by approximately 30-60% as compared to control. Enhanced proliferation under these conditions was confirmed using ATSCs isolated from three independent donors. ATSCs that were expanded in the presence of FGF-2 and DEX for 5 days were capable of differentiating into either osteoblastic or adipogenic cells, and the cells were positive for the mesenchymal stem cell markers such as CD29, CD44, CD90, CD105, and CD146, suggesting that the stem cell potential of the ATSCs was preserved. Analysis of signaling pathway revealed that tyrosine phosphorylation of Src kinase was dramatically increased in response to FGF-2 and DEX, suggesting the involvement of Src-dependent pathways in the stimulatory mechanism of proliferation of ATSCs by FGF-2 and DEX. Moreover, Src family kinase inhibitors (SU6656 and Src kinase inhibitor I) substantially reduced the FGF-2 and DEX induced proliferation of ATSCs. SU6656 also inhibited the osteogenic and adipogenic differentiation of ATSCs. The results of the current study demonstrate that FGF-2 in combination with DEX stimulates the proliferation and osteoblastic and adipogenic differentiation of ATSCs through a Src-dependent mechanism, and that FGF-2 and DEX promote the efficient ex vivo expansion of ATSCs. PMID- 19292684 TI - Formation of keratocyte spheroids on chitosan-coated surface can maintain keratocyte phenotypes. AB - Corneal keratocytes have been reported to be able to form spheroids that can preserve their phenotypes after being seeded back onto tissue culture plate in specific culture media. In this study, we found that keratocytes could also form spheroids on a bioengineered material, chitosan-coated surface, with 10% horse serum and Dulbecco's modified Eagle's medium. Under scanning electron microscopy observation, the cells in the spheroids were found to adhere each other tightly, and the cellular boundary could not be distinguished. They could return to a dendritic (keratocyte) morphology and proliferate after they were seeded back onto tissue culture plate. Immunocytochemistry was used to characterize these cells. Reverse transcription-polymerase chain reaction revealed that keratocytes in the spheroids were not from the PAX-6-positive progenitor cells. Further, the results of the seeding density and the number of spheroids formation, cell viability (MTT) assays, negative staining of Ki-67, and Live/Dead assay suggested that the spheroids were from cell aggregation instead of cell proliferation. Cells in the spheroids maintained phenotypes and functions characteristic of keratocytes, as seen by reverse transcription-polymerase chain reaction, collagen gel contraction assay, and challenges of keratocytes with transforming growth factor-beta1. Our results showed that corneal keratocytes could form spheroids on a chitosan-coated surface and maintain a keratocyte phenotype. However, such keratocyte spheroids do not proliferate and cannot withstand transforming growth factor-beta from myofibroblast differentiation. PMID- 19292685 TI - Occurrence and characterization of Salmonella Hiduddify from chickens and poultry meat in Nigeria. AB - The objective of this study was to determine the prevalence of Salmonella serovars and the antimicrobial susceptibility in chickens and poultry meat products in rural areas in Nigeria. The study was an observational cross sectional investigation in which the target population included exotic and local chickens in Maiduguri main markets, chickens from farms, and free-range local chickens. A total of 865 samples were collected from feces, kidney, lungs, cecum, intestine, liver, heart, gizzard, and cloacal swabs from 525 different chickens. Salmonella was isolated from 130 of the samples. A stratified random sampling technique was used to select 41 isolates out of the 130 strains for serotyping, pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing. Thirty-nine out of the 41 samples serotyped yielded Salmonella Hiduddify; two yielded a rough Salmonella serovar. The 39 Salmonella Hiduddify isolates and the two rough isolates were highly similar by PFGE typing, indicating that all of the isolates were of the same serovar. A low frequency of resistance was found among the isolates, except for resistance to ciprofloxacin for which 23 (56%) of the isolates tested exhibited resistance. This study documents for the first time the isolation of Salmonella Hiduddify in chickens and shows that this serovar is widespread in rural areas in Nigeria. It also documents a high frequency of fluoroquinone resistance in the isolates indicating the presence of selective pressure in the environment. Further studies should be conducted to reveal if the serovar is present in eggs and causes salmonellosis among the general population. PMID- 19292686 TI - Prevalence of antibiotic-resistant Salmonella in fish in Guangdong, China. AB - INTRODUCTION: This study examines fish from freshwater aquaculture operations in Guangdong Province, China, to determine the prevalence of antibiotic-resistant Salmonella isolates. This information can help identify risks of human exposure to Salmonella and guide decisions of whether to include farmed fish samples in routine food surveillance for Salmonella. METHODS: One hundred live freshwater farmed finfish were sampled from several stalls at two wholesale and four retail markets in Guangzhou from June to July 2008. Isolation and antibiotic sensitivity testing was done according to the U.S. FDA Bacteriological Analytical Manual. Antibiotic sensitivity testing was done using the Kirby Bauer disk diffusion method. RESULTS: All five Salmonella isolates were susceptible to neomycin, cefotaxime, and cefepime and resistant to erythromycin and penicillin. The most resistant isolate was susceptible to 7 of the 16 antibiotics tested. DISCUSSION: The estimated prevalence of Salmonella is 5% (95% CI: 2-11%) in live finfish from markets in Guangzhou, China. All five isolates were not susceptible to three or more antibiotics. Three of the five isolates had decreased susceptibility to nitrofurantoin, suggesting illegal use of nitrofurans in food animal production, and surveillance of resistance to this class of antibiotics is warranted. We suggest aquaculture-producing countries where there may be high antibiotic use to add farmed fish products to the list of foods they include in Salmonella surveillance. This would help evaluate human health risks posed by antibiotic resistant bacteria in farmed fish products. PMID- 19292687 TI - Emergence, distribution, and molecular and phenotypic characteristics of Salmonella enterica serotype 4,5,12:i:-. AB - Salmonella spp. represent one of the most common causes of bacterial foodborne illnesses around the world. The species Salmonella enterica contains more than 2500 serotypes, and emergence of new human pathogenic Salmonella strains and serotypes represents a major public health issue. Salmonella enterica subsp. enterica serotype 4,5,12:i:- represents a monophasic variant of Salmonella Typhimurium, which has rarely been identified before the mid-1990 s. The prevalence of this serotype among human salmonellosis cases has increased considerably since the mid-1990 s and Salmonella 4,5,12:i:- currently (i.e., the first decade of the 2000s) represents one of the most common serotypes among human cases in many countries around the world. This paper discusses our current knowledge of the global ecology, epidemiology, transmission, and evolution of this emerging Salmonella serotype. PMID- 19292688 TI - E. coli O157 from sheep in northeast Scotland: prevalence, concentration shed, and molecular characterization by multilocus variable tandem repeat analysis. AB - We report the prevalence, concentrations, and strain diversity of Escherichia coli O157 shed by sheep fed on root crops during a winter period in northeast Scotland. E. coli O157 was isolated on 6 farms from 14 studied during January to March 2005. The individual sheep prevalence was 5.8% and concentration excreted was <10(2) colony-forming units/g for all but one fecal sample. Verocytotoxigenic E. coli O157, determined by polymerase chain reaction and verocell assay, was recovered from 27% of samples. Four farms had sheep shedding the same strain as determined by multiple-locus variable analysis and no within-farm diversity was observed. The low numbers shed and the high levels of atoxigenic strains indicate a lower risk to human health from these animals compared to many ruminants grazing pasture during summer months. These data will be valuable for quantitative risk assessments and provide preliminary information that feeding sheep on root crops may be a practical intervention to reduce E. coli O157 infection in animals and ultimately humans. PMID- 19292689 TI - Characterization of pathogenic and resistant genome repertoire reveals two clonal lines in Salmonella enterica subsp. enterica serovar Paratyphi B (+)-tartrate positive. AB - A total of 36 contemporary human, animal, and environmental (+)-tartrate fermenting (dT+) Salmonella enterica serovar Paratyphi B isolates, formerly called Salmonella serovar Java, and five related monophasic S. enterica serovar 4,5,12:b:- isolates from Belgium, Germany, the Netherlands, and the United Kingdom were investigated for clonality and antimicrobial resistance profiles, as well as their virulence and resistance gene repertoire. Two major clonal lines, which could be phenotypically differentiated by the expression of the O:5 antigen, were identified. All O:5 antigen negative strains were multidrug resistant and originated (with two exceptions) from Belgian, Dutch, or German poultry. Strains exhibiting the O:5 antigen encoded by the oafA gene revealed a more heterogeneous group including multidrug-resistant and susceptible strains. Compared to O:5 antigen negative isolates, Salmonella Paratyphi B dT+ O:5 positive strains possessed additional virulence determinants. The Salmonella genomic island 1 was only found in O:5 positive strains. Five monophasic Salmonella 4,5,12:b:- lacking the phase-2 flagellar antigen were assigned to Salmonella Paratyphi B dT+ isolates of the O:5 positive group. The conclusion of the analysis is that Salmonella Paratyphi B dT+ O:5 negative and O:5 positive isolates evolved from a different lineage. Salmonella Paratyphi B dT+ O:5 positive strains possess additional fimbrial and virulence genes that probably enable this clone to interact with a broader range of hosts and the environment. Salmonella Paratyphi B dT+ O:5 negative continuously persists in poultry across Western Europe, especially Belgium, the Netherlands, and Germany. PMID- 19292690 TI - Modeling on-farm Escherichia coli O157:H7 population dynamics. AB - Escherichia coli O157:H7 is a potentially fatal foodborne pathogen with a putative reservoir for human infection in feedlot cattle. In order to more effectively identify targets for intervention strategies, we aimed to (1) assess the role of various feedlot habitats in E. coli O157:H7 propagation and (2) provide a framework for examining the relative contributions of animals and the surrounding environment to observed pathogen dynamics. To meet these goals we developed a mathematical model based on an ecological metapopulation framework to track bacterial population dynamics inside and outside the host. We used E. coli O157:H7 microbiological and epidemiological literature to characterize E. coli O157:H7 habitats at the pen level and account for E. coli O157:H7 population processes in water troughs, feedbunks, cattle hosts, and pen floors in the model. Simulations indicated that E. coli O157:H7 was capable of maintaining viable populations in the feedlot without net growth in the cattle gastrointestinal tract, suggesting E. coli O157:H7 may not always act as an obligate parasite. Water troughs and contaminated pen floors appeared to be particularly influential sources driving E. coli O157:H7 population dynamics and thus would serve as prime environmental targets for interventions to effectively reduce the E. coli O157:H7 load at the pen level. PMID- 19292691 TI - Development and use of a chromogenic macroarray system for the detection of Staphylococcus aureus with enterotoxin A, B, C, D, E, and G genes in food and milk samples. AB - Abstract Staphylococcus aureus is one of the major pathogens that can cause staphylococcal infection and food poisoning. There are five major classical types of staphylococcal enterotoxins (SEs): SEA, SEB, SEC, SED, and SEE, as well as new SEs or SE-like superantigens (SAgs), such as SEG to SEU. Since many S. aureus strains harbor more than one SE gene and identification of SEs involved in food poisoning cases is time consuming, we developed a chromogenic macroarray method that allows convenient and simultaneous detection of classical SE genes and a new SE gene (seg), which is phylogenetically highly related to seb and sec. Two sets of degenerated primers labeled with biotin were used to co-amplify all SE genes in S. aureus strains through the polymerase chain reaction (PCR). Afterwards, these biotin-labeled PCR products were hybridized with SE gene-specific probes spotted on the nitrocellulose membrane. When this macroarray was used to detect enterotoxingenic S. aureus in milk or beef homogenate containing 10(0)-10(4) target cells per milliliter or gram of the sample, all six enterotoxin genes could be identified after a 12-hour enrichment step. This macroarray offers clinical and food inspection laboratories a rapid and economical visual method to detect common enterotoxigenic S. aureus strains. PMID- 19292692 TI - Persistent infection of a carcinoma thyroid cell line with coxsackievirus B. AB - BACKGROUND: Viral infections are described as environmental factors that are implicated in various thyroid diseases. The role of enteroviruses (EV) in the pathogenesis of thyroid diseases has been suspected. Recently, we found that EV RNA could be detected in postoperative thyroid specimens. We decided to investigate the infection of a human thyroid cell line with coxsackievirus B4 (CVB4). METHODS: The wild-type human thyroid carcinoma cells K1 were inoculated with CVB4E2 at 2.1 x 10(7) TCID50/mL. The titer of the virus progeny was determined every 3 days on HEp-2 cells. CVB replication at the molecular level was monitored by searching for intracellular viral genomes using reverse transcription (RT)-polymerase chain reaction (PCR). EV VP1 capsid protein was detected by indirect immunocytofluorescence staining. Cell viability was determined by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) absorbance, and the nuclear morphology was assessed by Hoechst Dye staining. RESULTS: Infectivity assays with CVB4E2 revealed an increase in viral titers. Virus production decreased thereafter, but was not stopped by serial subculture for 24 days after infection. Detection of intracellular positive and negative viral RNA strands by RT-PCR was positive between days 1 and 14 postinfection (p.i.), and by semi-nested RT-PCR up to 24 days. K1 cell cultures infected with CVB4 were stained positively for EV VP1: the number of VP1-positive cells decreased rapidly within 6 days and remained low up to the end of culture. Compared with mock-infected cultures, viability in CVB4-infected cultures was around 100% up to 24 days. Cells with strongly fluorescent nuclei and/or fragmented nuclei were observed. CONCLUSION: We demonstrate for the first time that CVB4 could replicate in thyroid cells and could persist, with predominance of viral negative RNA strands for up to 24 days p.i. without obvious cytopathic effect. Our results suggest that CVB4 may lead to thyroid cell apoptosis. Further studies are needed to determine whether CVB could play a role in thyroid pathologies. PMID- 19292693 TI - Hypothermic therapies targeting brain and spinal cord injury. Introduction. PMID- 19292694 TI - A tribute to Dr. Hubert L. Rosomoff: a pioneer in treatment of pain and use of hypothermia. PMID- 19292695 TI - Posthypothermic rewarming considerations following traumatic brain injury. AB - To date, considerable attention has been focused upon the use of hypothermia as a therapeutic strategy for attenuating many of the damaging consequences of traumatic brain injury (TBI). Despite the promise of hypothermic intervention following TBI, many questions remain regarding the optimal use of hypothermic intervention, including, but not limited to, the rewarming rates needed to assure optimal brain protection. In this review, we revisit the relatively limited literature examining the issue of hypothermia and differing rewarming rates following TBI. Considering both experimental and clinical literature, evidence is presented that the rate of posthypothermic rewarming is an important variable for influencing the protective effects of hypothermic intervention following TBI. In the experimental setting, posttraumatic hypothermia followed by slow rewarming appears to provide maximal protection in terms of traumatically induced axonal damage, microvascular damage and dysfunction, and contusional expansion. In contrast, hypothermia followed by rapid rewarming not only reverses the protective effects associated with hypothermic intervention, but in many cases, exacerbates the traumatically induced pathology and its functional consequences. While similar evaluations have not been conducted in the clinical setting, multiple lines of clinical evidence suggest the benefits of posttraumatic hypothermia are optimized through the use of slow rewarming, with the suggestion that such a strategy reduces the potential for rebound vasodilation, elevated intracranial pressure (ICP), and impaired neurocognitive recovery. Collectively, this review highlights not only the benefits of hypothermic intervention, but also the rate of posthypothermic rewarming as an important variable in assuring maximal efficacy following the use of hypothermic intervention. PMID- 19292696 TI - Management of pitfalls for the successful clinical use of hypothermia treatment. AB - Therapeutic hypothermia is a promising method for controlling intracranial pressure (ICP) in severely brain-injured patients. However, clinical data regarding the effect of brain hypothermia on overall outcome of these patients is limited. This may be because there are specific pitfalls associated with the clinical management of induced hypothermia in patients with severe traumatic brain injury (TBI). These pitfalls may be avoided by preventing specific risk factors when cooling is induced and with rewarming. However, these risk factors have not been well systematically discussed in the literature. In this paper, three categories of clinical issues regarding the management of brain hypothermia are discussed: (1) stress-induced secondary brain injury mechanisms; (2) technical aspects of intensive care unit (ICU) cooling management; and (3) rewarming rates and methods. For patients with a Glasgow Coma Scale (GCS) score of less than 8, management of stress-induced insulin-resistant hyperglycemia, and unstable systemic circulation due to impaired cardiac contractility are especially important. For example, in our experience, posttraumatic hyperglycemia, exacerbated by cooling, may be ameliorated by the administration of a ketone body with mannitol. Prevention of selective free radical damage to neurons is also an important target for successful brain hypothermia treatment. Taken together, it is clear that several orchestrated steps should be initiated to enhance the protective effects of hypothermia therapy and prevent these possible pitfalls. PMID- 19292697 TI - Current and future role of therapeutic hypothermia. AB - Therapeutic moderate hypothermia has been advocated for use in traumatic brain injury, stroke, cardiac arrest-induced encephalopathy, neonatal hypoxic-ischemic encephalopathy, hepatic encephalopathy, and spinal cord injury, and as an adjunct to aneurysm surgery. In this review, we address the trials that have been performed for each of these indications, and review the strength of the evidence to support treatment with mild/moderate hypothermia. We review the data to support an optimal target temperature for each indication, as well as the duration of the cooling, and the rate at which cooling is induced and rewarming instituted. Evidence is strongest for prehospital cardiac arrest and neonatal hypoxic-ischemic encephalopathy. For traumatic brain injury, a recent meta analysis suggests that cooling may increase the likelihood of a good outcome, but does not change mortality rates. For many of the other indications, such as stroke and spinal cord injury, trials are ongoing, but the data are insufficient to recommend routine use of hypothermia at this time. PMID- 19292705 TI - First in line or first in time? Effects of settlement order and arrival date on reproduction in a group-living beetle Dendroctonus ponderosae. AB - 1. In group-living organisms, individuals that initiate aggregations, termed pioneers, may suffer higher mortality costs than individuals that join established aggregations. Here we examine the hypothesis that aggregation initiators achieve higher reproductive success in the early phases of colonization, potentially through lower competition and increased access to the resource (finder's advantage), and that this benefit is sufficient to outweigh the costs of pioneering. 2. We also examine the role of arrival date (irrespective of order within the aggregation) on reproductive success, because individuals in seasonal environments may gain an advantage by arriving early. We test these hypotheses using mountain pine beetles, Dendroctonus ponderosae (Coleoptera: Curculionidae, Scolytinae), an obligately aggregating insect wherein pioneers suffer high mortality due to tree defences. We measured reproductive success at the start of winter when most components of final offspring number were likely to be determined. 3. Surviving pioneers that successfully recruited conspecifics had smaller broods than individuals that joined aggregations, refuting our hypothesis. The later that a beetle settled within an aggregation, the higher its reproductive success. However, beetles that settled early in the season produced more offspring than those that settled later in the season, and this effect was generally stronger than settlement order within an aggregation. Our study highlights the importance of examining the effects of both settlement order and arrival date on the costs and benefits of pioneering aggregations. PMID- 19292706 TI - Assignment of 128 genes localized on human chromosome 14q to the IMpRH map. AB - To provide a gene-based comparative map and to examine a porcine genome assembly using bacterial artificial chromosome-based sequence, we have attempted to assign 128 genes localized on human chromosome 14q (HSA14q) to a porcine 7000-rad radiation hybrid (IMpRH) map. This study, together with earlier studies, has demonstrated the following. (i) 126 genes were incorporated into two SSC7 RH linkage groups by CarthaGene analysis. (ii) In the remaining two genes, TOX4 linked to TCRA located in SSC7 by two-point analysis, whereas SIP1 showed no significant linkage with any gene/marker registered in the IMpRH Web Server. (iii) In the two groups, the gene clusters located from 19.9 to 36.5 Mb on HSA14q11.2-q13.3 and from 64.0 to 104.3 Mb on HSA14q23-q32.33 respectively were assigned to SSC7q21-q26. (iv) Comparison of the gene order between the present RH map and the latest porcine sequence assembly revealed some inconsistencies, and a redundant arrangement of 16 genes in the sequence assembly. PMID- 19292707 TI - Development of gene-associated intronic TR markers for the Pacific abalone Haliotis discus hannai. PMID- 19292708 TI - Monophyletic origin of domestic bactrian camel (Camelus bactrianus) and its evolutionary relationship with the extant wild camel (Camelus bactrianus ferus). AB - The evolutionary relationship between the domestic bactrian camel and the extant wild two-humped camel and the factual origin of the domestic bactrian camel remain elusive. We determined the sequence of mitochondrial cytb gene from 21 camel samples, including 18 domestic camels (three Camelus bactrianus xinjiang, three Camelus bactrianus sunite, three Camelus bactrianus alashan, three Camelus bactrianus red, three Camelus bactrianus brown and three Camelus bactrianus normal) and three wild camels (Camelus bactrianus ferus). Our phylogenetic analyses revealed that the extant wild two-humped camel may not share a common ancestor with the domestic bactrian camel and they are not the same subspecies at least in their maternal origins. Molecular clock analysis based on complete mitochondrial genome sequences indicated that the sub-speciation of the two lineages had begun in the early Pleistocene, about 0.7 million years ago. According to the archaeological dating of the earliest known two-humped camel domestication (5000-6000 years ago), we could conclude that the extant wild camel is a separate lineage but not the direct progenitor of the domestic bactrian camel. Further phylogenetic analysis suggested that the bactrian camel appeared monophyletic in evolutionary origin and that the domestic bactrian camel could originate from a single wild population. The data presented here show how conservation strategies should be implemented to protect the critically endangered wild camel, as it is the last extant form of the wild tribe Camelina. PMID- 19292709 TI - Development of a 25-plex SNP assay for traceability in cattle. AB - Single nucleotide polymorphisms (SNPs) are amenable to automation and therefore have become the marker of choice for DNA profiling. SNaPshot, a primer extension based method, was used to multiplex 25 SNPs that have been previously validated as useful for identity control. Detection of extended products was based on four different fluorochromes and extension primers with oligonucleotide tails of differing lengths, thus controlling the concise length of the entire chromatogram to 81 bases. Allele frequencies for Holstein, Simmental, Limousin, Angus, Charolais and Tux Cattle were estimated and significant positive Pearson correlation coefficients were obtained among the analysed breeds. The probability that two randomly unrelated individuals would share identical genotypes for all 25 loci varied from 10(-8) to 10(-10) for these breeds. For parentage control, the exclusion power was found to be 99.9% when the genotypes of both putative parents are known. A traceability test of duplicated samples indicated a high genotyping precision of >0.998. This was further corroborated by analysis of 60 cases of parent-sib pairs and trio families. The 25-plex SNaPshot assay is adapted for low- and high-throughput capacity and thus presents an alternative for DNA-based traceability in the major commercial cattle breeds. PMID- 19292710 TI - Genome-wide genetic diversity of 'Nici', the DNA source for the CHORI-260 turkey BAC library and candidate for whole genome sequencing. AB - Vertebrate whole genome sequence assembly can benefit from a priori knowledge of variability in the target genome, with researchers often selecting highly inbred individuals for sequencing. However, for most species highly inbred research lines are lacking, requiring the use of an outbred individual(s). Here we examined the source DNA [Nicholas inbred (Nici)] of the CHORI-260 turkey bacterial artificial chromosome (BAC) library through analysis of microsatellites and BAC sequences. Heterozygosity of Nici was compared with that of individuals from several breeder lines. Seventy-eight microsatellites were screened for polymorphism in a total of 43 birds, identifying an average individual heterozygosity of 0.39, with Nici at 0.35. Additional loci (total of 147) were examined on a subset of individuals to obtain better genome coverage. The mean heterozygosity for this subset was 0.33 with Nici at 0.31. Examination of approximately 200 kb of genome sequence identified SNPs in the order of one per 200 bp in Nici. These data suggest that the heterozygosity of Nici is comparable to other birds of selected breeder lines and that whole genome sequencing would result in an abundant resource of genome-wide polymorphisms. PMID- 19292711 TI - Mitochondrial DNA diversity and origins of South and Central American goats. AB - We have analysed the genetic diversity of South and Central American (SCA) goats by partially sequencing the mitochondrial control region of 93 individuals with a wide geographical distribution. Nucleotide and haplotype diversities reached values of 0.020 +/- 0.00081 and 0.963 +/- 0.0012 respectively. We have also observed a rather weak phylogeographic structure, with almost 69% of genetic variation included in the within-breed variance component. The topology of a median-joining network analysis including 286 European, Iberian, Atlantic and SCA mitochondrial sequences was very complex, with most of the haplotypes forming part of independent small clusters. SCA sequences showed a scattered distribution throughout the network, and clustering with Spanish and Portuguese sequences occurred only occasionally, not allowing the distinguishing of a clear Iberian signature. Conversely, we found a prominent cluster including Canarian, Chilean, Argentinian and Bolivian mitochondrial haplotypes. This result was independently confirmed by constructing a Bayesian phylogenetic tree (posterior probability of 0.97). Sharing of mitochondrial haplotypes by SCA and Canarian goats suggests that goat populations from the Atlantic archipelagos, where Spanish and Portuguese ships en route to the New World used to stow food and supplies, participated in the foundation of SCA caprine breeds. PMID- 19292712 TI - A case of delayed-type hypersensitivity to subcutaneously administered drugs during anticancer immunotherapy. PMID- 19292713 TI - Cytarabine-associated lymphoid dyscrasia with atypical T CD30+ infiltrate in a patient suffering from acute nonlymphoblastic leukaemia. PMID- 19292714 TI - A therapeutic effect of STAT6 decoy oligodeoxynucleotide ointment in atopic dermatitis: a pilot study in adults. PMID- 19292715 TI - Primary cutaneous CD4+ small/medium-sized pleomorphic T-cell lymphoma associated with an annular elastolytic giant cell granuloma. PMID- 19292716 TI - Switching between antitumour necrosis factor alpha biologic agents--is patient weight an important consideration? PMID- 19292717 TI - Audit of dermatological content of U.K. undergraduate curricula. AB - BACKGROUND: Recommendations for the dermatology content (learning outcomes) of the core undergraduate curriculum were sent to all U.K. medical schools in June 2006. OBJECTIVE: To carry out an audit of the content of the core curriculum in each U.K. medical school against the recommendations for a core undergraduate dermatology curriculum (the criteria) published by the British Association of Dermatologists, to identify areas of good practice and to gather evidence for developing the learning and teaching of dermatology. METHODS: A questionnaire was circulated to the dermatology teaching leads of all U.K. medical schools (29) and one Irish medical school. Questions which the teaching leads were unable to answer were sent to the relevant deans and responses incorporated into the results. All curricula should include the essential learning outcomes that focus on clinical skills; as this was an audit to benchmark current practice, we did not set standards for the other recommendations for a core curriculum. RESULTS: Replies were received from teaching leads in 29 of the 30 medical schools and from 16 of the deans. Essential clinical skills such as taking a dermatological history and examining the skin were included in the curricula of most, but not all, medical schools. Areas of good practice include teaching on tumours, acne and psoriasis, but we found some surprising omissions including the diagnosis of meningococcaemia. Our data suggest that some students have little exposure to dermatology, but dermatology teaching takes place in secondary care in all medical schools. Knowledge-based assessments are used by 27 medical schools. CONCLUSIONS: Curricula should be strengthened so that the recommended learning outcomes feature in the core curricula of all medical schools. Teaching leads in all specialties, including those in the community, should communicate so that learning and teaching are integrated horizontally and vertically. The results should provide a baseline for future audits. PMID- 19292718 TI - A novel 3017-bp deletion mutation in the FERMT1 (KIND1) gene in a Chinese family with Kindler syndrome. PMID- 19292719 TI - PORCN gene mutations and the protean nature of focal dermal hypoplasia. AB - Focal dermal hypoplasia (FDH) is an X-linked dominant disorder featuring developmental abnormalities of ectodermal and mesodermal tissues. Pathogenic mutations in the PORCN gene (locus Xp11.23) were identified in 2007 and thus far 27 different mutations have been reported. PORCN encodes a putative O acyltransferase which facilitates secretion of Wnt proteins required for ectomesodermal tissue development. We investigated PORCN gene pathology and pattern of X-chromosome inactivation analysis in two unrelated Caucasian female patients who presented with multiple developmental abnormalities consistent with FDH. We also reviewed the clinical and molecular data for all reported PORCN mutations and assessed genotype-phenotype correlation for sporadic and familial cases of FDH. DNA sequencing revealed two new PORCN gene mutations: p.W282X and c.74delG (p.G25fsX51). X-chromosome inactivation analysis revealed a random pattern in one case but was uninformative in the other. Collectively, point/small mutations account for 24 out of the 29 PORCN mutations and are typically seen in sporadic cases; larger deletions are more common in familial cases. Identification of two new PORCN gene mutations confirms the importance of PORCN associated Wnt signalling in embryogenesis. Both new cases showed Blaschko-linear dermal hypoplasia and extensive ectomesodermal abnormalities, including severe limb developmental anomalies and a giant cell tumour of bone in one patient. Clinical variability can be attributed to the degree of lyonization and postzygotic genomic mosaicism, which are important mechanisms in determining the clinical presentation. PMID- 19292720 TI - Mutations in the P2RY5 gene underlie autosomal recessive hypotrichosis in 13 Pakistani families. AB - BACKGROUND: Autosomal recessive hypotrichosis is a rare genetic irreversible hair loss characterized by sparse scalp hair, sparse to absent eyebrows and eyelashes, and sparse axillary and body hair. Affected male individuals have normal beard hair. OBJECTIVES: To search for pathogenic mutations in the human P2RY5 gene in Pakistani families with autosomal recessive hereditary hypotrichosis. METHODS: In the present report, 16 unrelated consanguineous Pakistani families having multiple affected individuals with autosomal recessive hypotrichosis were investigated. Linkage in these families was searched by genotyping microsatellite markers linked to autosomal recessive hypotrichosis loci LAH1, LAH2 and LAH3. Thirteen of the families showed linkage to the LAH3 locus on chromosome 13q14.11 q21.32. These families were then subjected to direct sequencing of the P2RY5 gene, which encodes a G protein-coupled receptor. RESULTS: Sequence analysis of the P2RY5 gene revealed two novel missense mutations (c.742A>T; p.N248Y and c.830C>T; p.L277P) in three families. Five previously described mutations including three missense (c.188A>T; p.D63V, c.436G>A; p.G146R, c.562A>T; p.I188F), one insertion (c.69insCATG; p.24insHfsX52) and one complex deletion (c.172-175delAACT; 177delG; p.N58-L59delinsCfsX88) were detected in the other 10 families. CONCLUSIONS: Mutations revealed in the present study extend the body of evidence implicating the P2RY5 gene in the pathogenesis of human hereditary hair loss. PMID- 19292732 TI - Immunohistochemical detection of polyductin and co-localization with liver progenitor cell markers during normal and abnormal development of the intrahepatic biliary system and in adult hepatobiliary carcinomas. AB - The longest open reading frame of PKHD1 (polycystic kidney and hepatic disease 1), the autosomal recessive polycystic kidney disease (ARPKD) gene, encodes a single-pass, integral membrane protein named polyductin or fibrocystin. A fusion protein comprising its intracellular C-terminus, FP2, was previously used to raise a polyclonal antiserum shown to detect polyductin in several human tissues, including liver. In the current study, we aimed to investigate by immunohistochemistry the detailed polyductin localization pattern in normal (ductal plate [DP], remodelling ductal plate [RDP], remodelled bile ducts) and abnormal development of the primitive intrahepatic biliary system, known as ductal plate malformation (DPM). This work also included the characterization of polyductin expression profile in various histological forms of neonatal and infantile cholestasis, and in cholangiocellular carcinoma (CCC) and hepatocellular carcinoma (HCC). We detected polyductin expression in the intrahepatic biliary system during the DP and the RDP stages as well as in DPM. No specific staining was found at the stage of remodelled bile ducts. Polyductin was also detected in liver biopsies with neonatal cholestasis, including mainly biliary atresia and neonatal hepatitis with ductular reaction as well as congenital hepatic fibrosis. In addition, polyductin was present in CCC, whereas it was absent in HCC. Polyductin was also co-localized in some DP cells together with oval stem cell markers. These results represent the first systematic study of polyductin expression in human pathologies associated with abnormal development of intrahepatic biliary tree, and support the following conclusions: (i) polyductin expression mirrors developmental properties of the primitive intrahepatic biliary system; (ii) polyductin is re-expressed in pathological conditions associated with DPM and (iii) polyductin might be a potential marker to distinguish CCC from HCC. PMID- 19292733 TI - Renal, vascular and cardiac fibrosis in rats exposed to passive smoking and industrial dust fibre amosite. AB - Passive smoking is an independent risk factor for cardiovascular diseases. Industrial fibrous dust, e.g. the asbestos group member, amosite, causes lung cancer and fibrosis. No data are available on renal involvement after inhalational exposure to these environmental pollutants or of their combination, or on cardiovascular and renal toxicity after exposure to amosite. Male Wistar rats were randomized into four groups (n= 6): control and amosite group received initially two intratracheal instillations of saline and amosite solution, respectively. Smoking group was subjected to standardized daily exposure to tobacco smoke for 2 hrs in a concentration resembling human passive smoking. Combined group was exposed to both amosite and cigarette smoke. All rats were killed after 6 months. Rats exposed to either amosite or passive smoking developed significant glomerulosclerosis and tubulointerstitial fibrosis. Combination of both exposures had additive effects. Histomorphological changes preceded the clinical manifestation of kidney damage. In both groups with single exposures, marked perivascular and interstitial cardiac fibrosis was detected. The additive effect in the heart was less pronounced than in the kidney, apparent particularly in changes of vascular structure. Advanced oxidation protein products, the plasma marker of the myeloperoxidase reaction in activated monocytes/macrophages, were increased in all exposed groups, whereas the inflammatory cytokines did not differ between the groups. In rats, passive smoking or amosite instillation leads to renal, vascular and cardiac fibrosis potentially mediated via increased myeloperoxidase reaction. Combination of both pollutants shows additive effects. Our data should be confirmed in subjects exposed to these environmental pollutants, in particular if combined. PMID- 19292734 TI - HER-2 overexpression/amplification in Barrett's oesophagus predicts early transition from dysplasia to adenocarcinoma: a clinico-pathologic study. AB - Barrett's oesophagus (BO) is the primary precursor lesion for oesophageal adenocarcinoma (ADC). The natural history of metaplasia-dysplasia-carcinoma sequence remains largely unknown. HER2/neu oncogene results overexpressed/amplified in preneoplastic lesions and in ADC of the oesophagus and it has been associated with poor prognosis. Our aim was to evaluate the role of HER2 overexpression/amplification in predicting the conversion from precursor lesions to ADC. We retrospectively evaluated by univariate analysis of single variables clinical records and histological specimens of 21 patients with a confirmed diagnosis of BO and/or oesophageal dysplasia. Clinical variables included age, gender, alcohol and smoking intake, presence of symptoms (pyrosis, disphagia) and endoscopic features (length). HER2 status was studied by immunohistochemistry and fluorescence in situ hybridization (FISH) on paraffin embedded tissue. The end-points were the occurrence of progression and the time to-progression (TTP) from the initial histologic lesion to the worst pathological pattern. Median age at diagnosis was 63 years (range 37-84). BO median length was 4.5 cm. Progression occurred in 11 of 21 patients and median TTP was 24 months. HER2 was overexpressed/amplified in 8 of 21 (38%) patients. HER2 overexpression/ amplification and the presence of dysplasia were statistically associated with progression (P= 0.038). This study provides evidence for a possible role of HER2 in the transition from dysplasia to ADC of the oesophagus. This fact could help in identifying patients at high risk of malignant transformation. PMID- 19292744 TI - Reporting of participant compliance in randomized controlled trials of nutrition supplements during pregnancy. AB - Randomized controlled trials (RCT) are widely considered to be the gold standard for demonstrating intervention effects. Adequacy of reporting of participant compliance in RCTs affects the interpretation of study results. Our aims were two fold: first, to assess the adequacy of reporting of participant compliance in RCTs investigating the effect of maternal nutritional supplements on infant outcomes; and second, to examine authors' adherence to the Consolidated Standards of Reporting Trials (CONSORT) guidelines on participant flow. Papers reporting trials of nutritional supplementation during pregnancy, and published after revision of the CONSORT recommendations, were identified using a search of medical databases. Two researchers systematically reviewed the papers to assess the reporting of participant compliance according to specified criteria, and the presentation of participant flow data recommended in the CONSORT guidelines. The literature search identified 58 papers. Almost a third (n = 18) did not describe how participant compliance was assessed. Nearly half of the papers (n = 27) failed to report participant compliance numerically (absolute numbers or percentage) and differences in compliance data between treatment arms were not reported in 52% of papers (n = 28). The majority (83%) gave no information on whether the study protocol included any researcher input aimed at maximizing compliance. In addition to inadequate reporting of compliance, two of the CONSORT requirements (eligibility criteria and numbers discontinuing the intervention) were inadequately reported in 69% and 60% of papers, respectively. We conclude that participant compliance in nutrition trials is frequently inadequately reported. 'False negative' results from RCTs with poor compliance could wrongly influence policy and inhibit further research concerned with nutritional supplementation for women of child-bearing age. We suggest that changes to the CONSORT guidelines may improve RCT reporting. PMID- 19292735 TI - Insights into cerebrovascular complications and Alzheimer disease through the selective loss of GRK2 regulation. AB - Alzheimer disease (AD) and stroke are two leading causes of age-associated dementia. Increasing evidence points to vascular damage as an early contributor to the development of AD and AD-like pathology. In this review, we discuss the role of G protein-coupled receptor kinase 2 (GRK2) as it relates to individuals affected by AD and how the cardiovasculature plays a role in AD pathogenesis. The possible involvement of GRKs in AD pathogenesis is an interesting notion, which may help bridge the gap in our understanding of the heartbrain connection in relation to neurovisceral damage and vascular complications in AD, since kinases of this family are known to regulate numerous receptor functions both in the brain, myocardium, and elsewhere. The aim of this review is to discuss our findings of overexpression of GRK2 in the context of the early pathogenesis of AD, because increased levels of GRK2 immunoreactivity were found in vulnerable neurons of AD patients as well as in a two-vessel occlusion (2-VO) mammalian model of ischaemia. Also, we consider the consequences for this overexpression as a loss of G-protein coupled receptor (GPCR) regulation, as well as suggest a potential role for GPCRs and GRKs in a unifying theory of AD pathogenesis, particularly in the context of cerebrovascular disease. We synthesize this newer information and attempt to put it into context with GRKs as regulators of diverse physiological cellular functions that could be appropriate targets for future pharmacological intervention. PMID- 19292745 TI - Plasma folate status and dietary folate intake among Chinese women of childbearing age. AB - Maternal folic acid deficiency is an underlying risk for neural tube defects (NTDs). China has one of the highest prevalences of NTDs, and the prevalence rates of NTDs vary by region. We characterized plasma folate level and dietary folate intake among Chinese women of childbearing age by region (North and South, East and West, urban and rural) to provide evidence for establishing policy to prevent NTDs. A total of 1003 women of childbearing age from five provinces in China were interviewed. Fasting blood samples were collected. Plasma folate concentrations were determined by a microbiological assay. Dietary intake data were collected using a 24-h recall. Both the plasma folate concentrations and dietary folate intake of women in the South (25.9 nmol L(-1) and 211.0 microg day(-1)) were higher than those of women in the North (13.3 nmol L(-1) and 189.2 microg day(-1)). In the North, plasma folate concentrations and dietary folate intake of women in rural areas were lower than those of women in urban areas, whereas, in the South, an opposite pattern was observed. No difference was found between women in the East and West, in either the North or South regions. Plasma folate and dietary folate intake among Chinese women of childbearing age were suboptimal and varied by region. Different folic acid supplementation approaches and dosage should be undertaken to improve folate status of women in different areas. Particular attention should be paid to women in the North, especially in northern rural areas. PMID- 19292746 TI - Parental distress around supplementing breastfed babies using nasogastric tubes on the post-natal ward: a theme from an ethnographic study. AB - There is abundant evidence of the benefits of breastfeeding. In the UK, supplementation in hospital has consistently been shown to be associated with shortened duration of breastfeeding. This paper reports on a subset of the data from an ethnographic study that explored the expectations, beliefs and experiences of mothers and health professionals concerning supplementation, using a variety of methods, of breastfed babies in an English maternity unit in 2002. This paper aims to describe the expectations, beliefs and experiences of mothers and health professionals concerning supplementation by nasogastric (NG) tube on the post-natal ward. Participant observation was carried out on day and night shifts and at weekends over 9 months. Mothers, midwives, neonatal nurses, health care assistants and senior paediatricians were interviewed. Categories and themes were generated. The researchers' constructs of 'the essential method', when the tube was the method needed for medical reasons, and 'the chosen method', when other methods of oral feeding should have been possible, emerged. The latter included time pressures and the avoidance of any form of oral activity that might perhaps make return to the breast more difficult. The data concerning the use of NG tubes for supplementation yielded the specific theme of parental distress. In the absence of evidence that supplementation by NG tube on the post-natal ward is associated with greater breastfeeding success than other methods, the use of the tube to avoid any form of 'oral confusion' should be discontinued. Its use primarily to save time should not be considered acceptable. PMID- 19292747 TI - Reliability of reported breastfeeding duration among reproductive-aged women from Mexico. AB - Breastfed children have lower risk of infectious diseases, post-neonatal mortality and chronic diseases later in life. Because epidemiologic studies usually rely on reported history of previous breastfeeding, data on the accuracy and precision of recalled histories allow improved interpretation of the epidemiologic findings. We evaluated the reliability of two reported breastfeeding durations in 567 reproductive-aged women from Mexico using information obtained from nearly identical sets of questions applied at different times after weaning. We compared differences between reports, and examined the intraclass correlation coefficient (ICC) for any and for exclusive breastfeeding (EBF). Logistic regression was used to evaluate the determinants of poor recall (difference between reports of >20%). The reliability of duration of any breastfeeding was high (ICC 0.94). Overall, differences between reports of duration were usually <1 month, and for 385/567, the difference was < or =0.5 months. Predictors of poorer recall were having > or =4 children, and time between reports of >2 months. The only predictor of better recall was greater age of the baby at weaning. The reliability of EBF duration was lower (ICC 0.49). In this population with a relatively long duration of breastfeeding, reliability of any breastfeeding duration was high. Age, education and previous breastfeeding were not important predictors of recall, in contrast to findings in earlier studies. Consistent with previous reports, however, parity and length of recall were associated with poorer recall of duration of any breastfeeding. Future studies that use reported breastfeeding duration may want to consider the effect of these variables on recall. PMID- 19292748 TI - Women's experiences of infant feeding support in the first 6 weeks post-birth. AB - Research suggests women find the first 2 to 6 weeks to be the most difficult time for breastfeeding. It has been identified that women need and seek support with breastfeeding during this time. Support is a difficult concept to define. When discussed by professionals, support for breastfeeding is generally viewed in terms of providing information and educational interventions. There is little understanding of the different elements of breastfeeding support strategies and the mechanisms by which support operates. Further, there is a paucity of qualitative research specifically reporting women's experiences and expectations of professional support. This paper describes women's expectations and experiences of 'infant feeding support' provided by health professionals in the first 6 weeks post-birth. The findings are drawn from a grounded theory study exploring women's infant feeding decisions in the first 6 weeks post-birth. Participants were recruited from a variety of socio-demographic areas of Sydney and the NSW Central Coast, Australia in 2003-2004. The women in this study discussed aspects of what they considered helpful and/or unhelpful in terms of professional support. In addition, they also provided insight into aspects of interactions that were deemed important to them as new mothers learning to feed their babies. The results are presented in three sections: expecting support, experiencing support and evaluating support. The findings help to better understand components of professional practices and behaviours that can be considered supportive. The support behaviours are far more complex than simply increasing education and knowledge of infant feeding. They demonstrate the need for sensitive individualized care and show that this type of support can increase women's confidence to breastfeed. PMID- 19292749 TI - Micronutrient sprinkles add more bioavailable iron to some Kenyan complementary foods: studies using an in vitro digestion/Caco-2 cell culture model. AB - Iron deficiency anaemia is arguably the most important public health problem in developing countries. In Sub-Saharan Africa, iron deficiency anaemia has largely been attributed to poor infant feeding practices where complementary foods low in iron bioavailability are offered to at-risk infants. Home fortification of complementary foods using micronutrient Sprinkles has been shown to reduce iron deficiency anaemia in many resource-poor settings. In this study, the benefit of using the micronutrient Sprinkles as a home fortificant for some Kenyan complementary foods was assessed using an in vitro Caco-2 cell model. In each case when micronutrient Sprinkles were added to the complementary food, the amount of Caco-2 cell ferritin formation increased. For example, the addition of Sprinkles to corn porridge increased ferritin formation 5-fold from 5.8 to 31.8 ng mg(-1). Therefore, as indicated by the results, micronutrient Sprinkles would be a suitable form of home fortification for Kenyan complementary foods. Their use should be encouraged so as to improve infant feeding practices in Kenya. PMID- 19292750 TI - Supplementary feeding with fortified spread among moderately underweight 6-18 month-old rural Malawian children. AB - We aimed to analyse growth and recovery from undernutrition among moderately underweight ambulatory children receiving micronutrient-fortified maize-soy flour (Likuni Phala, LP) or ready-to-use fortified spread (FS) supplementary diet. One hundred and seventy-six 6-18-month-old individuals were randomized to receive 500 g LP or 350 g FS weekly for 12 weeks. Baseline and end of intervention measurements were used to calculate anthropometric gains and recovery from underweight, wasting and stunting. Mean weight-for-age increased by 0.22 (95% CI 0.07-0.37) and 0.28 (0.18-0.40) Z-score units in the LP and FS groups respectively. Comparable increase for mean weight-for-length was 0.39 (0.20-0.57) and 0.52 (0.38-0.65) Z-score units. Recovery from underweight and wasting was 20% and 93% in LP group and 16% and 75% in FS group. Few individuals recovered from stunting and mean length-for-age was not markedly changed. There were no statistically significant differences between the outcomes in the two intervention groups. In a poor food-security setting, underweight infants and children receiving supplementary feeding for 12 weeks with ready-to-use FS or maize-soy flour porridge show similar recovery from moderate wasting and underweight. Neither intervention, if limited to a 12-week duration, appears to have significant impact on the process of linear growth or stunting. PMID- 19292751 TI - Cognitive restraint, uncontrolled eating and emotional eating: correlations between parent and adolescent. AB - The purpose of this study was to examine, in a general population, the resemblance in eating behaviour between adolescents and their parents. This study was based on the first examination of a community-based epidemiological study in Northern France. Subjects were offspring aged 14-22 years (135 boys and 125 girls) and their parents (174 fathers and 205 mothers). The Three-Factor Eating Questionnaire Revised 18-item version (TFEQ-R18) identified three aspects of eating behaviour: cognitive restraint of eating, uncontrolled eating and emotional eating. Familial resemblance in eating behaviour was measured by partial Spearman's correlations, adjusted for age and body mass index. Sons' uncontrolled eating was positively related to fathers' cognitive restraint of eating (r = 0.36), but not to fathers' uncontrolled eating (r = 0.07), nor to mothers' eating behaviour. Sons' cognitive restraint of eating was related to no parental eating behaviour scores. In daughters, cognitive restraint of eating was positively related to mothers' uncontrolled eating (r = 0.26), but not to mothers' cognitive restraint of eating (r = 0.13). Daughters' uncontrolled eating and emotional eating were positively associated with the same scores in mothers. Finally, daughters' eating behaviour was not related to fathers' eating behaviour. In conclusion, correlations in eating behaviour were higher with the parent of the same gender, and eating behaviours in adolescents seem to reflect opposition to parents' behaviour more than familial resemblance. PMID- 19292752 TI - Effect of parity on phalangeal bone mineral density in post-menopausal Sri Lankan women: a community based cross-sectional study. AB - There is paucity of studies related to parity and bone mineral density in South Asian countries. We recruited 713 healthy, community dwelling post-menopausal women from seven provinces in Sri Lanka for this survey. The number of pregnancies, including miscarriages beyond 20 weeks of gestation, was recorded. Women with diseases and those who have taken drugs that can affect bone mineral density (BMD) were excluded (n = 15). Phalangeal BMD and bone mineral content (BMC) were measured using AccuDEXA in 713 women. Mean (SE) BMD of nulliparous women (n = 32), women with one to two pregnancies (n = 284), three to four pregnancies (n = 290) and more than four pregnancies (n = 107) were 0.437(0.014), 0.454(0.005), 0.455(0.005) and 0.417(0.006) g/cm(2), respectively (P < 0.001). Corresponding mean (SE) BMCs were 1.30(0.063), 1.41(0.021), 1.43(0.022) and 1.32(0.033) g, respectively (P < 0.001). Women with more than four pregnancies were older and lighter when compared with other groups. When results were adjusted for current age and current weight, differences in mean BMD and BMC between groups became non-significant. BMD of nulliparous women remained low in all analyses. We report a significant difference in unadjusted phalangeal BMD in women categorized according to their parity. Women with one to four pregnancies had the highest phalangeal BMD and BMC, while multi-parous (more than four pregnancies) and nulliparous women had lower values. However, in an adjusted analysis, the differences in BMD and BMC were partially explained by the differences of age and body weight between the groups and the unique effect of parity was difficult to determine. Women with lower BMD may have a higher risk of future fractures. PMID- 19292753 TI - Vitamin D insufficiency common in newborns, children and pregnant women living in Newfoundland and Labrador, Canada. AB - Vitamin D deficiency is associated with poor bone health, colorectal cancer, type 1 diabetes and multiple sclerosis. Two national health-related societies in Canada have made recommendations for vitamin D supplementation, yet little research has been reported on the vitamin D status of Canadians. Lifestyle changes, such as sunscreen use, spending less time outdoors and insufficient intake of vitamin D-containing foods as well as northern latitude, may be affecting human vitamin D status. A cross-sectional analysis of 25-hydroxyvitamin D [25-(OH)D] was conducted in pregnant women, newborns (umbilical cord blood) and children. Samples were analysed by liquid chromatography mass spectrometry. Published ranges for 25-(OH)D were used to determine vitamin D status. The prevalence of 25-(OH)D deficiency for the three groups studied revealed most concentrations in the 25-(OH)D deficiency or insufficiency ranges. There were significant differences in all groups studied between seasons, with the exception of maternal blood and female cord blood samples. 25-(OH)D insufficiency was common in all groups for winter and summer, more so in winter. 25-(OH)D insufficiency was common in the three groups studied. The Newfoundland and Labrador population may be at increased risk for vitamin D insufficiency because of factors such as northern latitude and lifestyle issues. Further research on the vitamin D status of this population is important, considering the potential adverse health-related outcomes and the recommendations on supplementation being made. PMID- 19292755 TI - Neuropsychological and functional MRI studies provide converging evidence of anterior language dysfunction in BECTS. AB - PURPOSE: Benign childhood epilepsy with centrotemporal spikes (BECTS) is the most common epilepsy syndrome of childhood and can be associated with language difficulties. The exact profile of these difficulties and their neurofunctional underpinnings, however, are not yet clear. METHODS: To further understand the impact of the BECTS syndrome on language, we assessed language performance using standard neuropsychological measures, and patterns of language lateralization using functional magnetic resonance imaging (fMRI) in children with typical BECTS (n = 20) and healthy controls (n = 20). RESULTS: The fMRI analyses revealed that language-related activation was less lateralized to the left hemisphere in anterior brain regions in the patients relative to the control group. This finding was consistent with decreased performance in the BECTS group compared to the control group on the neuropsychological measure most dependent on the integrity of anterior aspects of the language axis, namely, sentence production. DISCUSSION: The converging lines of evidence from the neuropsychological and activation methodologies support the view that BECTS is associated with language difficulties that are regional, and anterior, in nature. PMID- 19292756 TI - Increased leak conductance in dentate gyrus granule cells of temporal lobe epilepsy patients with Ammon's horn sclerosis. AB - PURPOSE: Temporal lobe epilepsy (TLE) is often associated with Ammon's horn sclerosis (AHS) characterized by hippocampal cell death and dentate gyrus granule cell dispersion (GCD). Granule cells survive AHS and have been proposed to be hyperexcitable in TLE. Here we studied whether the passive excitability of granule cells correlates with the severity of AHS. METHODS: We analyzed the passive membrane properties of identified granule cells using patch-clamp recordings in acute tissue slices obtained from TLE surgery. Independent Wyler grading and GCD measurements were used to assess the severity of AHS. RESULTS: The input resistances and membrane time constants of granule cells were reduced in high-grade versus low-grade AHS samples and negatively correlated with the degree of GCD. Granule cells possessed large Ba(2+)-sensitive, inwardly rectifying K(+) conductances. DISCUSSION: The increased leak conductance, likely mediated by K(+) channels, does not argue for an increased excitability of granule cells but rather points to a neuroprotective mechanism in the sclerotic focus in TLE. PMID- 19292757 TI - Long-term antiepileptic drug therapy contributes to the acceleration of atherosclerosis. AB - PURPOSE: Long-term antiepileptic drug (AED) therapy has been associated with an increase in risk of atherosclerosis. At issue is whether this risk is related to the duration of AED therapy. We evaluated the hypothesis that the cumulative effect of long-term exposure to AEDs plays a pivotal role in the pathogenesis of atherosclerosis in patients with epilepsy. METHODS: One hundred ninety-five patients under long-term AED therapy and 195 healthy age- and sex-matched control subjects received measurement of intima media thickness (IMT) at the far wall of the common carotid artery (CCA) by B-mode ultrasonography to assess the extent of atherosclerosis. Other measurements included body mass index (BMI) and blood lipid profile or homocysteine, folic acid, uric acid, fasting blood sugar, high sensitivity C-reactive protein (hs-CRP), thiobarbituric acid reactive substances (TBARS), and total reduced thiols. RESULTS: CCA IMT was significantly increased in patients with epilepsy, with male subjects exhibiting thicker IMT than their female counterparts. Whereas BMI, homocysteine, hs-CRP, and TBARS were significantly elevated, folic acid and thiols were significantly reduced in patients with epilepsy. Multiple linear regression analysis further revealed that duration of AED therapy, age, gender, and TBARS level (index for oxidative stress) were independently associated with CCA IMT. In addition, the log transformed CCA IMT increased linearly with duration of AED therapy after adjustments for age, gender, and TBARS level. DISCUSSION: The duration of AED therapy is significantly associated with the acceleration of atherosclerosis in patients with epilepsy, alongside independent contributions of age, gender, and oxidative stress to the atherosclerotic process. PMID- 19292758 TI - Does a SCN1A gene mutation confer earlier age of onset of febrile seizures in GEFS+? AB - SCN1A is the most clinically relevant epilepsy gene and is associated with generalized epilepsy and febrile seizure plus (GEFS+) and Dravet syndrome. We postulated that earlier onset of febrile seizures in the febrile seizure (FS) and febrile seizure plus (FS+) phenotypes may occur in the presence of a SCN1A mutation. This was because of the age-related onset of Dravet syndrome, which typically begins in the first year of life. We found that patients with FS and FS+ with SCN1A mutations had earlier median onset of febrile seizures compared to the population median. Patients with GABRG2 mutations had a similar early onset in contrast to patients with SCN1B mutations where onset was later. This study is the first to demonstrate that a specific genetic abnormality directly influences the FS and FS+ phenotype in terms of age of onset. PMID- 19292759 TI - Automated normalized FLAIR imaging in MRI-negative patients with refractory focal epilepsy. AB - BACKGROUND: Patients with focal epilepsy that is refractory to medical treatment are often considered candidates for resective surgery. Magnetic resonance imaging (MRI) has a very important role in the presurgical work-up of these patients, but is unremarkable in about one-third of cases. These patients are often deferred from surgery or have a less positive outcome if surgery is eventually undertaken. The aim of this study was to evaluate our recently described voxel-based technique using routine T2-FLAIR (fluid-attenuated inversion-recovery) scans in MRI-negative patients and to compare the results with video-EEG (electroencephalography) telemetry (VT) findings. METHODS: We identified 70 epilepsy patients with refractory focal seizures who underwent VT and had a normal routine MRI. T2-FLAIR scans were bias-corrected, and intensity and spatially normalized (nFSI) using Statistical Parametric Mapping 5 (SPM5) as previously described. Individual scans were then compared against a set of 25 normal controls using a voxel-based method. RESULTS: SPM5 identified 10 patients with suprathreshold clusters (14.3%). In 50% of these there was concordance between the lobe of the most significant cluster and the presumed lobe of seizure onset, as defined by VT. All cases were concordant with respect to lateralization of the putative focus. CONCLUSION: Using nFSI we identified focal structural cerebral abnormalities in 11.4% of patients with refractory focal seizures, and normal conventional MRI, that were fully or partially concordant with scalp VT. This voxel-based analysis of FLAIR scans, which are widely available, could provide a useful tool in the presurgical evaluation of epilepsy patients. Ongoing work is to compare these imaging findings with the results of intracranial EEG and histology of surgical resections. PMID- 19292760 TI - Involvement of snapdragon benzaldehyde dehydrogenase in benzoic acid biosynthesis. AB - Benzoic acid (BA) is an important building block in a wide spectrum of compounds varying from primary metabolites to secondary products. Benzoic acid biosynthesis from L-phenylalanine requires shortening of the propyl side chain by two carbons, which can occur via a beta-oxidative pathway or a non-beta-oxidative pathway, with benzaldehyde as a key intermediate. The non-beta-oxidative route requires benzaldehyde dehydrogenase (BALDH) to convert benzaldehyde to BA. Using a functional genomic approach, we identified an Antirrhinum majus (snapdragon) BALDH, which exhibits 40% identity to bacterial BALDH. Transcript profiling, biochemical characterization of the purified recombinant protein, molecular homology modeling, in vivo stable isotope labeling, and transient expression in petunia flowers reveal that BALDH is capable of oxidizing benzaldehyde to BA in vivo. GFP localization and immunogold labeling studies show that this biochemical step occurs in the mitochondria, raising a question about the role of subcellular compartmentalization in BA biosynthesis. PMID- 19292761 TI - Peroxule extension over ER-defined paths constitutes a rapid subcellular response to hydroxyl stress. AB - Plants survive against myriad environmental odds while remaining rooted to a single spot. The time scale over which plant cells can respond to environmental cues is seldom appreciated. Fluorescent protein-assisted live imaging of peroxisomes reveals that they respond within seconds of exposure to hydrogen peroxide and hydroxyl radicals by producing dynamic extensions called peroxules. Observations of the Arabidopsis flu mutant and treatments with xenobiotics eliciting singlet oxygen and superoxide reactive oxygen species suggest that the observed responses are specific for hydroxyl radicals. Prolonged exposure to hydroxyl radicals inhibits peroxule extension, and instead causes motile and spherical peroxisomes in a cell to become immotile and elongate several-fold. Expression of photo-convertible EosFP-PTS1 demonstrates that vermiform peroxisomes result from rapid stretching of individual peroxisomes, while the subsequent 'beads-on-a-string' morphology results from differential protein distribution within an elongated tubule. Over time, the beads in elongated peroxisomes also extend peroxules randomly before undergoing asynchronous, asymmetrical fission. Peroxule extension does not appear to involve cytoskeletal elements directly, but is closely aligned with and reflects the dynamics of ER tubules. Peroxisomal responses reveal a rapidly invoked subcellular machinery that is involved in recognition of hydroxyl stress thresholds, and its possible remediation locally through extension of peroxules or globally by increasing peroxisome numbers. A matrix protein retro-flow mechanism that supports peroxisome-ER connectivity in plant cells is suggested. PMID- 19292762 TI - Tandem affinity purification and mass spectrometric analysis of ubiquitylated proteins in Arabidopsis. AB - Protein ubiquitylation is a central regulatory mechanism that controls numerous processes in plants, including hormone signaling, developmental progression, responses to biotic and abiotic challenges, protein trafficking and chromatin structure. Despite data implicating thousands of plant proteins as targets, so far only a few have been conclusively shown to be ubiquitylated in planta. Here we describe a method to isolate ubiquitin-protein conjugates from Arabidopsis that exploits a stable transgenic line expressing a synthetic poly-UBQ gene encoding ubiquitin (Ub) monomers N-terminally tagged with hexahistidine. Following sequential enrichment by Ub-affinity and nickel chelate-affinity chromatography, the ubiquitylated proteins were trypsinized, separated by two dimensional liquid chromatography, and analyzed by mass spectrometry. Our list of 54 non-redundant targets, expressed by as many as 90 possible isoforms, included those predicted by genetic studies to be ubiquitylated in plants (EIN3 and JAZ6) or shown to be ubiquitylated in other eukaryotes (ribosomal subunits, elongation factor 1alpha, histone H1, HSP70 and CDC48), as well as candidates whose control by the Ub/26S proteasome system is not yet appreciated. Ub attachment site(s) were resolved for a subset of these proteins, but surprisingly little sequence consensus was detected, implying that specific residues surrounding the modified lysine are not important determinants for ubiquitylation. We also identified six of the seven available lysine residues on Ub itself as Ub attachment sites, together with evidence for a branched mixed-linkage chain, suggesting that the topologies of Ub chains can be highly complex in plants. Taken together, our method provides a widely applicable strategy to define ubiquitylation in any tissue of intact plants exposed to a wide range of conditions. PMID- 19292763 TI - The A-chain of insulin is a hot-spot for CD4+ T cell epitopes in human type 1 diabetes. AB - Type 1 diabetes (T1D) is caused by T cell-mediated destruction of the pancreatic insulin-producing beta cells. While the role of CD4(+) T cells in the pathogenesis of T1D is accepted widely, the epitopes recognized by pathogenic human CD4(+) T cells remain poorly defined. None the less, responses to the N terminal region of the insulin A-chain have been described. Human CD4(+) T cells from the pancreatic lymph nodes of subjects with T1D respond to the first 15 amino acids of the insulin A-chain. We identified a human leucocyte antigen-DR4 restricted epitope comprising the first 13 amino acids of the insulin A-chain (A1 13), dependent upon generation of a vicinal disulphide bond between adjacent cysteines (A6-A7). Here we describe the analysis of a CD4(+) T cell clone, isolated from a subject with T1D, which recognizes a new HLR-DR4-restricted epitope (KRGIVEQCCTSICS) that overlaps the insulin A1-13 epitope. This is a novel epitope, because the clone responds to proinsulin but not to insulin, T cell recognition requires the last two residues of the C-peptide (Lys, Arg) and recognition does not depend upon a vicinal disulphide bond between the A6 and A7 cysteines. The finding of a further CD4(+) T cell epitope in the N-terminal A chain region of human insulin underscores the importance of this region as a target of CD4(+) T cell responses in human T1D. PMID- 19292764 TI - Tumour necrosis factor-alpha processing in interstitial lung disease: a potential role for exogenous proteinase-3. AB - Tumour necrosis factor (TNF) blockade has become an important immunomodulatory therapy, particularly in patients refractory to conventional immunosuppression, but responses can be unpredictable. Understanding the complex biology of TNF processing may be key to predicting such responses and reduce unwanted side effects. TNF bioavailability is regulated partly by TNF-alpha converting enzyme (TACE) cleavage; however, it can also be cleaved by proteinase-3 (PR-3). We have demonstrated this mechanism previously in healthy human alveolar macrophages (AM), leading us to hypothesize that PR-3-mediated TNF processing may be an important mechanism in inflammatory lung disease. Furthermore, this may be more apparent in diseases with a neutrophil component typical of usual interstitial pneumonia (UIP), compared with sarcoidosis, where lymphocytes predominate. We isolated AM from patients with UIP and sarcoidosis and healthy subjects. We found increased TACE expression on AM in sarcoidosis. In contrast, TACE was not increased in UIP; we found increased cleavage of glutathione S-transferase proTNF) substrate, relative to both sarcoidosis and healthy controls. Furthermore, cleavage was subject to inhibition by serine protease inhibitor, rather than a TACE inhibitor BB-3103. Cleavage was proportional to the number of neutrophils isolated from bronchoalveolar lavage, whereas there was an inverse relationship between neutrophils and BB-3103 inhibition. There was also increased PR-3 on the AM surface in UIP relative to healthy controls. These data provide evidence for PR-3-mediated cleavage in UIP, which may have implications for future therapeutic targeting of TACE. PMID- 19292765 TI - Oral glucocorticoid treatment decreases interleukin-10 receptor expression on peripheral blood leucocyte subsets. AB - Glucocorticoids (GCS) are capable of stimulating the secretion of interleukin (IL)-10 by leucocytes; however, the potential of GCS to modulate leucocyte susceptibility to IL-10-mediated actions has not yet been studied. In the current paper, we performed a detailed cross-sectional analysis of IL-10 receptor (IL 10R) expression by CD4(+) and CD8(+) T cells, natural killer (NK) cells, monocytes and neutrophils. Next, we analysed the effects of short-term oral GCS treatment on surface IL-10R expression by various leucocyte subpopulations in asthmatic patients. All leucocyte subsets studied presented with substantial levels of surface IL-10R. The highest levels of IL-10R were found on monocytes, predominantly with CD14(2+)CD16(+) and CD14(+)CD16(+) phenotypes, and on CD4(+)CD25(high) T cells. In contrast, levels of IL-10R on CD8(+) T cells, NK cells and neutrophils were significantly lower and similar to each other in intensity. GCS treatment resulted in a significant decrease of IL-10R expression on all analysed peripheral blood leucocyte subsets. Our data suggest that down regulation of IL-10R could counterbalance the otherwise suppressive action of GCS. PMID- 19292766 TI - The expression profile of functional regulatory T cells, CD4+CD25high+/forkhead box protein P3+, in patients with ulcerative colitis during active and quiescent disease. AB - Regulatory T cells (T(reg)) have an essential role in maintaining immune tolerance in the gut. The functional CD4(+) T(reg) express the transcription factor forkhead box protein 3 (FoxP3) or a CD25(high) in humans. Further, depletion of elevated granulocytes/monocytes by extracorporeal adsorption (GMA) induces immunomodulation in patients with ulcerative colitis (UC). We investigated the impact of GMA on T(reg). Thirty-one UC patients, clinical activity index (CAI) 12.1 +/- 2.97, refractory to conventional medications including intravenous corticosteroid and 13 healthy controls (HC), were included. Patients received five GMA sessions over 5 weeks. Biopsies from the rectal mucosa and blood samples at baseline and post-GMA were immunostained with anti-CD4/FoxP3 and anti-CD4/CD25 antibodies for immunohistochemistry and flow cytometry. Following GMA, 22 of 31 patients achieved remission (CAI 5 mm in size. Twenty-five percent of patients had extra colonic abnormalities. There were no recorded complications. CONCLUSION: Our CTC practice is within accepted published guidelines. Bowel preparation is suboptimal in a significant proportion of cases and faecal tagging is being implemented. PMID- 19292777 TI - Secretory breast carcinoma in a 63-year-old man. AB - A secretory carcinoma of the breast, associated with axillary node metastasis, occurred in a 63-year-old man. Secretory breast carcinoma is a rare variant of breast carcinoma, but it is the commonest type seen in children, although most of the cases have been described in females. Here, we describe a case in an adult man. PMID- 19292778 TI - The impact of temperature change on the activity and community composition of sulfate-reducing bacteria in arctic versus temperate marine sediments. AB - Arctic regions may be particularly sensitive to climate warming and, consequently, rates of carbon mineralization in warming marine sediment may also be affected. Using long-term (24 months) incubation experiments at 0 degrees C, 10 degrees C and 20 degrees C, the temperature response of metabolic activity and community composition of sulfate-reducing bacteria were studied in the permanently cold sediment of north-western Svalbard (Arctic Ocean) and compared with a temperate habitat with seasonally varying temperature (German Bight, North Sea). Short-term (35)S-sulfate tracer incubations in a temperature-gradient block (between -3.5 degrees C and +40 degrees C) were used to assess variations in sulfate reduction rates during the course of the experiment. Warming of arctic sediment resulted in a gradual increase of the temperature optima (T(opt)) for sulfate reduction suggesting a positive selection of psychrotolerant/mesophilic sulfate-reducing bacteria (SRB). However, high rates at in situ temperatures compared with maximum rates showed the predominance of psychrophilic SRB even at high incubation temperatures. Changing apparent activation energies (E(a)) showed that increasing temperatures had an initial negative impact on sulfate reduction that was weaker after prolonged incubations, which could imply an acclimatization response rather than a selection process of the SRB community. The microbial community composition was analysed by targeting the 16S ribosomal RNA using catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH). The results showed the decline of specific groups of SRB and confirmed a strong impact of increasing temperatures on the microbial community composition of arctic sediment. Conversely, in seasonally changing sediment sulfate reduction rates and sulfate-reducing bacterial abundance changed little in response to changing temperature. PMID- 19292780 TI - Dermal contributions to UVA-induced oxidative stress in skin. AB - BACKGROUND: When the skin is exposed to solar irradiation, UVA photons interact with skin tissues and induce excessive reactive oxygen species, resulting in oxidative stress. We have shown in a previous study that in vivo chemiluminescence's measurement can be used to evaluate the overall level of UVA induced oxidative stress in human skin. However, the origin of the observed chemiluminescence signals remains unclear. METHODS: UVA-induced chemiluminescence measurements were conducted: (a) in vitro on collagen solutions and solid collagen sheet preparations, (b) ex vivo on human and mouse skin biopsies, and (c) in vivo on human skin of various constitutive pigmentation levels. Fluorescence was measured on collagen in vitro as well as on skin for the in vivo experiments. RESULTS: We found in the in vitro experiments that UVA-induced chemiluminescence increases with the presence of collagen cross-links. When dermal sides were exposed to UVA irradiation, both mouse and human skin biopsies demonstrated significantly higher chemiluminescence levels than when epidermal sides were exposed to UVA. The amount of collagen cross-links decreases slightly following UVA exposure, as shown both by in vivo fluorescence and by UVA-induced chemiluminescence. Finally, there was less measurable UVA-induced chemiluminescence in dark skin compared with light pigmented skin in vivo. CONCLUSIONS: The dermis is very sensitive to UVA photons. Dermal cross-links are potential UVA sensitizers. The oxidative stress induced by UVA and measured by chemiluminescence may largely be attributed to the breakdown of dermal collagen cross-links. PMID- 19292781 TI - Pinpoint papular polymorphous light eruption in Asian skin: a variant in darker skinned individuals. AB - BACKGROUND: Polymorphous light eruption (PMLE) is the most common idiopathic but probably immunologic photodermatosis and has wide morphological variants. METHODS: The photobiological features of all patients diagnosed with the pinpoint papular variant of PMLE at a tertiary dermatology centre in Singapore over a five year period were retrospectively examined. RESULTS: Twenty-one patients were reviewed from 2003 to 2007. There were 11 (52.4%) Chinese, four (19%) Malays, five (23.8%) Indians and one (4%) Cambodian. 14 (66.7%) were males and seven (33.3%) were females. The face/neck (48%) and arms/forearms (95%) were most often affected. Nineteen (90.5%) had Fitzpatrick skin phototype IV and two (9.5%) had skin phototype V. Six (28.6%) had decreased minimal erythema dose (MED) to ultraviolet B (UVB) light only, one (4.8%) had decreased MED to ultraviolet A (UVA) light only and one had decreased MED to both UVA and UVB. Four patients had photoprovocation test done, of which three had positive testing to UVA and one had negative testing to both UVA and UVB. Two histological subtypes were found in our patients, one showing perivascular dermatitis and the other consistent with lichen nitidus. CONCLUSION: Our data suggest that pinpoint papular PMLE is not uncommon in darker-skinned individuals in our cohort. PMID- 19292782 TI - Effects of near-infrared laser exposure in a cellular model of wound healing. AB - BACKGROUND: Clinical studies have demonstrated beneficial outcomes for low-level laser therapy (LLLT) using near-infrared (NIR) wavelengths. It has been hypothesized that the benefits of NIR LLLT are due in part to the thermal effects of NIR exposure. However, it is not clear whether photochemical interactions between NIR light and superficial tissues contribute to beneficial outcomes. To investigate the photochemical effects of NIR exposure, the efficacy of 980 nm NIR LLLT on human fibroblast growth rates is investigated using an in vitro model of wound healing. METHODS: A small pipette is used to induce a wound in fibroblast cell cultures, which are imaged at specific time intervals over 48 h and exposed to a range of laser doses (1.5-66 J/cm(2)) selected to encompass the range of doses used during other in vivo and in vitro studies. For each image acquired, wound sizes were quantified using a novel application of existing image processing algorithms. RESULTS: Cell growth rates were compared across different laser exposure intensities with the same exposure duration, and across different laser exposure durations with the same exposure intensity. Exposure to low- and medium-intensity laser light accelerates cell growth, whereas high-intensity light negated the beneficial effects of laser exposure. Cell growth was accelerated over a wide range of exposure durations using medium-intensity laser light, with no significant inhibition of cell growth at the longest exposure durations used in this study. CONCLUSION: Low-level exposure to 980 nm laser light can accelerate wound healing in vitro without measurable temperature increases. However, these results also demonstrate the need for appropriate supervision of laser therapy sessions to prevent overexposure to NIR laser light that may inhibit cell growth rates observed in response to lower intensity laser exposure. PMID- 19292783 TI - Spectrophotometric intracutaneous analysis as an early non-invasive predictor of efficacy in the phototherapy of psoriasis. AB - BACKGROUND: Phototherapy is generally effective for psoriasis but individual responsiveness and optimal treatment duration for disease clearance are unpredictable. However, easy, rapid and non-invasive plaque assessment by spectrophotometric intracutaneous analysis (SIAscopy), a novel multispectral skin imaging technique, may now make prediction feasible. OBJECTIVES: The early prediction of psoriatic plaque clearance during phototherapy by SIAscopy. METHODS: Sixteen psoriatic plaques in 10 psoriasis patients were serially assessed SIAscopically during phototherapy for punctate dots representing the dilated papillary dermal blood vessels characteristic of active psoriasis, and the results compared with the clinical findings. RESULTS: All plaques showing full SIAscopic clearance at early follow-up also showed complete or almost complete clinical clearance, and remained the same thereafter. All showing no SIAscopic clearance at early follow-up showed at most partial clinical clearance, and also remained the same thereafter. All showing only partial SIAscopic clearance at early follow-up also showed just partial clinical clearance, but then generally progressed to full SIAscopic and clinical clearance. CONCLUSIONS: SIAscopy of psoriatic plaques at early follow-up during patient phototherapy enables good prediction of likely later clinical clearance, thereby potentially avoiding unnecessary further treatment. A larger confirmatory study is now needed. PMID- 19292784 TI - Tanning salons in Santiago, Chile: the knowledge of the staff in charge and the quality of information provided to potential clients before and after a new regulatory law. AB - INTRODUCTION: In recent years, the use of artificial tanning devices has become very popular in Chile. In April, 2007, a new law was established to regulate the non-medical use of tanning beds and tanning booths. The aim of this study was to evaluate the level and quality of information provided by tanning salon staff to clients before deciding on using the services. This information was collected before and 6 months after the law was enacted by researchers visiting tanning salons posing as potential clients. METHODS: Using a previously designed questionnaire, 24 tanning salons in Santiago, Chile, were evaluated by researchers acting as potential customers without experience with artificial tanning devices. Employees were questioned using a hidden questionnaire before and 6 months after enactment of the law. RESULTS: Before the law, 25% of the establishments gave spontaneous information about the risks of using tanning beds, which increased to 45.8% after enactment of the law. Clients never received written information, either before or after enactment of the law. On both evaluations, more than 1/3 of the employees affirmed that there were no risks in using tanning beds. Before enactment of the law, only 12.5% of the tanning salons required the use of goggles, while after the law, this increased to 25% (P=NS). Only two establishments required that clients sign an informed consent form after enactment of the law. DISCUSSION: The level of knowledge among employees in tanning salons was very low and the quality of the information they provided to the clients was very poor. It was important to legislate on ultraviolet tanning services, but at present, the impact of the law has been very limited. PMID- 19292785 TI - Failure of short-term psoralen and ultraviolet A light maintenance treatment to prevent early relapse in patients with chronic recurring plaque-type psoriasis. AB - BACKGROUND: Photochemotherapy using psoralen and ultraviolet A light (PUVA) is a highly effective treatment option for patients with severe psoriasis. Maintenance treatment has been advocated to provide for sustained remission. However, only a few studies have been conducted to assess the efficacy of maintenance treatment and these have provided inconsistent results. METHODS: We performed a prospective intrapatient left-right comparison study in 34 patients with chronic relapsing plaque psoriasis. PUVA treatment for clearing was given four times weekly. After complete or near-complete clearing, all patients were placed on a halfside maintenance schedule with irradiation twice weekly and then once weekly for 4 weeks each. The psoriasis area and severity index score was determined at baseline, end of the clearing phase and at 2-monthly intervals after discontinuation of treatment. RESULTS: Using a short-term maintenance protocol, a moderate delay in relapse of psoriasis was observed in only three patients (8.8%; 95% CI: 1.8-23.6%). In the remaining patients (91.2%), maintenance treatment had no effect on the length of remission. The mean time interval until relapse without and with maintenance irradiation was 4.5 +/- 3.4 and 4.6 +/- 3.4 months, respectively. CONCLUSION: Our data indicate that short-term maintenance treatment is not effective in preventing early relapse of psoriasis and should be avoided. PMID- 19292786 TI - Indoor tanning in North Rhine-Westphalia Germany: a self-reported survey. AB - BACKGROUND/AIMS: We examined the prevalence of indoor tanning in North Rhine Westphalia and identified correlates of sunbed use. METHODS: During regular skin cancer screening campaigns 1242 subjects completed and returned a structured questionnaire on constitutional parameters and indoor tanning habits. RESULTS: The regular sunbed user rate (more than 10 exposures/year) was 15.4% (191/1242). Most sunbed users were under 29 years of age. The number of female sunbed users was greater than the number of male users. Respondents with secondary modern school qualification used sunbeds more infrequently than respondents with junior high school and high school qualifications. Respondents with skin type III and IV used sunbeds more frequently. Tanning and preparation for sunny holidays were the main reasons for sunbed use. The most frequently reported positive effects experienced by means of sunbed use were improved appearance and relaxation. Most respondents indicated that they hardly or never had sunburns following indoor tanning. Almost half of respondents consider radiation generated by sunbeds somewhat dangerous. CONCLUSIONS: Use of indoor tanning in North Rhine-Westphalia has increased and is significantly associated with female gender, younger age, high-level education, and skin type III and IV. The motivation for sunbed use and benefits experienced are mainly based on the perception of improved appearance due to tan and increased sense of well-being. PMID- 19292787 TI - Oral vs. bath PUVA using 8-methoxypsoralen for chronic palmoplantar eczema. AB - BACKGROUND: Both oral and bath PUVA with 8-methoxypsoralen (8-MOP) have been shown to be effective in the treatment of chronic palmoplantar eczema. However, most studies were retrospective and did not include longer follow-up periods. AIM: To compare the therapeutic efficacy, tolerability and duration of remission after oral vs. bath PUVA using 8-MOP in patients with chronic palmoplantar eczema. METHODS: Twenty-nine patients were randomly allocated to treatment with oral or bath PUVA. Treatment was given thrice weekly for a maximum of 20 weeks. The primary outcome measure was the improvement in eczema score at the end of treatment. After clearing patients were followed up until relapse or up to 40 months. RESULTS: Overall, both PUVA modalities appeared comparably effective. However, after stratifying according to eczema type, significant differences in therapeutic outcome in general as well as in response to the two regimes were found. Dyshidrotic eczema responded better to both treatments (P=0.048) and remained longer in remission than hyperkeratotic eczema. Hyperkeratotic eczema cleared significantly better with oral than with bath PUVA (P=0.03). CONCLUSION: Oral PUVA is preferable for patients with hyperkeratotic eczema and bath PUVA for patients with dyshidrotic eczema. PMID- 19292788 TI - p53 protein expression in skin with different levels of photoaging. AB - In non-melanoma skin cancer, keratinocytes show marked p53 protein expression. Considering photoaging as a background for skin cancer, p53 protein expression by immunohistochemistry was evaluated in normal skin with different levels of photoaging. The difference in p53 expression between mild and severe levels of photoaging was not statistically significant (P=0.169). PMID- 19292789 TI - The effects of phototherapy on the numbers of circulating natural killer cells and T lymphocytes in psoriasis. AB - The innate immune system is believed to be important in the pathogenesis of psoriasis and natural killer (NK) have been found in increased numbers in psoriatic plaques. Alterations in the numbers of NK cells in peripheral blood have been reported. We investigated the effect of phototherapy on levels of peripheral NK cells and lymphocytes in patients with psoriasis. In nine patients whom we followed before, during and after narrowband ultraviolet B (UVB) treatment there were no differences in the numbers of circulating lymphocytes, lymphocyte subsets or cells expressing NK markers and controls. Treatment with narrowband UVB did, however, significantly lower circulating CD4 counts which gradually recovered posttreatment. PMID- 19292790 TI - Response of vulval lichen sclerosus and squamous hyperplasia to photodynamic treatment using sustained topical delivery of aminolevulinic acid from a novel bioadhesive patch system. AB - This study evaluated the clinical and histopathological responses of vulval lichen sclerosus (LS) and squamous hyperplasia (SH) to photodynamic therapy (PDT). A novel bioadhesive patch containing aminolevulinic acid (ALA) at a dose of (38 mg/cm(2)) was used to treat 10 patients before irradiation with light of 630 nm. Clinical, histopathological and pathological responses to treatment were assessed at 6 weeks post-treatment. After 17 cycles of PDT, six patients reported significant symptomatic relief and no cutaneous photosensitivity. Histopathological differences were not demonstrated, but statistically significant induction of apoptosis was seen. It can be concluded that ALA-PDT patch-based formulation is pragmatic and primarily offers symptomatic management of vulval LS and SH. PMID- 19292791 TI - The spontaneous resolution of photosensitivity and contact allergy in a patient with chronic actinic dermatitis. AB - Chronic actinic dermatitis (CAD) is an uncommon photodermatosis in which patients typically exhibit photosensitivity together with contact allergy. Improvement of the photosensitivity over time is well recognized in a proportion of patients but the concurrent resolution of contact allergy is rare. We report on a 46-year-old female with longstanding CAD who demonstrated resolution of both of these features. PMID- 19292792 TI - A link between the North Atlantic Oscillation and measles dynamics during the vaccination period in England and Wales. AB - Ecologists have become aware of the role played by interannual climatic variability on the temporal dynamics of infectious diseases. In this report, I present evidence from data on measles cases in England and Wales showing that during the post-vaccination period, the interannual variability of winter weather (represented by the North Atlantic Oscillation, NAO) influences the annual dynamics of the disease. Using annual measles data from seven cities and simple logistic models, this study reveals how, after vaccination, NAO increases its effects on measles fluctuations. In addition, this study shows that vaccination may be represented as a simple vertical and lateral perturbation effect (Royama's classification), by reducing the maximum per capita growth rate and the equilibrium number of infected individuals. The results suggest that vaccination will not lead to outbreaks of measles from regular cyclic to irregular chaotic dynamics. In contrast, because of the reduction in per capita growth rates, the disease dynamics appear to be more stable than during the pre-vaccination period. The analysis of annual data on infectious diseases may be useful for detecting long-term effects of climate and complements the classical analyses and modeling based on monthly or seasonal time-step data. PMID- 19292793 TI - Climatic drivers of hemispheric asymmetry in global patterns of ant species richness. AB - Although many taxa show a latitudinal gradient in richness, the relationship between latitude and species richness is often asymmetrical between the northern and southern hemispheres. Here we examine the latitudinal pattern of species richness across 1003 local ant assemblages. We find latitudinal asymmetry, with southern hemisphere sites being more diverse than northern hemisphere sites. Most of this asymmetry could be explained statistically by differences in contemporary climate. Local ant species richness was positively associated with temperature, but negatively (although weakly) associated with temperature range and precipitation. After contemporary climate was accounted for, a modest difference in diversity between hemispheres persisted, suggesting that factors other than contemporary climate contributed to the hemispherical asymmetry. The most parsimonious explanation for this remaining asymmetry is that greater climate change since the Eocene in the northern than in the southern hemisphere has led to more extinctions in the northern hemisphere with consequent effects on local ant species richness. PMID- 19292794 TI - Mechanistic niche modelling: combining physiological and spatial data to predict species' ranges. AB - Species distribution models (SDMs) use spatial environmental data to make inferences on species' range limits and habitat suitability. Conceptually, these models aim to determine and map components of a species' ecological niche through space and time, and they have become important tools in pure and applied ecology and evolutionary biology. Most approaches are correlative in that they statistically link spatial data to species distribution records. An alternative strategy is to explicitly incorporate the mechanistic links between the functional traits of organisms and their environments into SDMs. Here, we review how the principles of biophysical ecology can be used to link spatial data to the physiological responses and constraints of organisms. This provides a mechanistic view of the fundamental niche which can then be mapped to the landscape to infer range constraints. We show how physiologically based SDMs can be developed for different organisms in different environmental contexts. Mechanistic SDMs have different strengths and weaknesses to correlative approaches, and there are many exciting and unexplored prospects for integrating the two approaches. As physiological knowledge becomes better integrated into SDMs, we will make more robust predictions of range shifts in novel or non-equilibrium contexts such as invasions, translocations, climate change and evolutionary shifts. PMID- 19292796 TI - Therapeutic decisions for ductal carcinoma in situ: a Gordian knot. PMID- 19292797 TI - Prognostic significance of oncogenic markers in ductal carcinoma in situ of the breast: a clinicopathologic study. AB - Ductal carcinoma in situ (DCIS) is a heterogeneous malignant condition of the breast with an excellent prognosis. Until recently mastectomy was the standard treatment. As the results of the National Surgical Adjuvant Breast and Bowel Project-17 trial and the introduction of the Van Nuys Prognostic Index (VNPI) less radical therapies are used. Objectives are to identify clinicopathologic and biologic factors that may predict outcome. Cases of DCIS diagnosed in two Belgian University Centers were included. Paraffin-embedded material and Hematoxylin and Eosin stained slides of DCIS cases were reviewed and tumor size, margin width, nuclear grade, and comedo necrosis were assessed. Molecular markers (estrogen receptor, progesterone receptor, HER1-4, Ki67, and c-myc) were assayed immunohistochemically. Applied treatment strategies were correlated with the prospective use of the VNPI score. Kaplan-Meier survival plots were generated with log-rank significance and multiple regression analysis was carried out using Cox proportional hazards regression analysis; 159 patients were included with a median age of 54 years (range 29-78); 141 had DCIS and 18 DCIS with microinvasion. The median time of follow-up was 54 months (range 5-253). Twenty three patients developed a recurrence (14.5%). The median time to recurrence was 46 months (range 5-253). Before the introduction of the VNPI, 37.5% of the DCIS patients showed a recurrence while thereafter 6.7% recurred (p < 0.005). Two recurrences occurred in the VNPI group I (7.1%); seven in the VNPI group II (8.5%) (median time to recurrence 66.3 months) and 14 in the VNPI group III (28.5%) (median time to recurrence 40.2 months) (disease-free survival [DFS]: p < 0.05). A Cox proportional hazards regression analysis indicated that tumor size, margin width, pathologic class, and age were independent predictors of recurrence, but none of the studied molecular markers showed this. Overexpression of HER4 in the presence of HER3 was found to be associated with a better DFS (p < 0.05). This study confirms the value of the VNPI score and questions the benefit of an aggressive approach in the low-risk DCIS lesions. Independent predictors for recurrence included size, margin width, pathologic class, and age, but none of the molecular markers were part of it. Overexpression of HER4 in the presence of HER3 was associated with a better DFS. PMID- 19292798 TI - Early detection of breast cancer through population-based mammographic screening in Asian women: a comparison study between screen-detected and symptomatic breast cancers. AB - The first nation-wide mammographic screening program in Asia, BreastScreen Singapore (BSS), was launched in Singapore in January 2002. This study compared the presentation and results of screen-detected breast cancers with symptomatic breast cancers in two affiliated high-volume institutions, one of which was an assessment centre for BSS. The medical records of patients diagnosed with primary breast cancer at the Department of General Surgery, Singapore General Hospital and the Department of Surgical Oncology, National Cancer Centre, Singapore, during the period January 2002 to December 2003 were reviewed. Clinical and pathological comparisons were made between screen-detected lesions and symptomatic lesions. Of a total of 767 cases, 640 (83.4%) were invasive carcinomas and 127 (16.6%) were ductal carcinoma in-situ (DCIS) lesions. Only 13.4% of them were screen-detected. Compared to symptomatic cancers, screen detected lesions were of smaller size (median size 18 versus 23 mm), a lower stage (stages 0-2, 95 versus 83.2%) and histologic grade (grade 1-2, 71 versus 60%), with a higher incidence of DCIS (31.0 versus 14.3%) and had higher rates of breast conservation (45.6 versus 28.2%) (all p-values <0.05). By multivariate analysis, tumor palpability, tumor size >20 mm, nodal involvement, cerbB2 overexpression, and advanced disease stage were independent poor prognostic factors for disease-free survival, whereas nodal involvement, advanced disease, and recurrence predicted poor cancer-specific survival. However, there was no statistically significant difference in disease-free survival or cancer-specific survival between the two groups at a median follow-up of 38 months. Screening mammography has allowed the detection of smaller and hence oncologically more favorable lesions in Asian women. Although no significant survival benefit was demonstrated in our study, a longer period of follow-up is essential before the benefit of mortality reduction, as a result of mammography screening becomes evident in our population. PMID- 19292799 TI - Intraoperative high-dose-rate brachytherapy--a novel technique in the surgical management of axillary recurrence. AB - Treating recurrent disease in the axilla is a challenging and complex clinical problem. Several reports in the literature suggest better outcomes with the combination of both surgery and radiation therapy than either modality alone. However, the available options for re-treatment are limited by the extent of disease at relapse, and the prior therapy that the patient has already received. The choice of re-irradiation using conventional external beam therapy is generally limited because of the risk of exceeding the radiation tolerance of the brachial plexus. In lieu of our concerns regarding excessive morbidity from re irradiation with external radiation therapy when treating an axillary relapse, we applied intraoperative high-dose-rate brachytherapy (HDR-IORT) at the time of surgical resection +/- a modest dose of postoperative external beam radiation therapy. In this paper, we describe the feasibility of HDR-IORT technique in three patients presenting with recurrent disease in the axilla. PMID- 19292800 TI - Should histologic type be taken into account when considering neoadjuvant chemotherapy in breast carcinoma? AB - Neoadjuvant chemotherapy is becoming the standard of care in locally advanced breast cancers. With complete pathologic response, patients may have a better overall survival. However, most patients do not have a complete pathologic response, and it is unclear how this impacts survival and whether histologic subtype or chemotherapeutic histologic changes play a role. We retrospectively identified 49 cases of invasive breast carcinoma treated with neoadjuvant chemotherapy (40 ductal, nine lobular) and examined histologic and biologic features of ductal and lobular carcinoma before and after chemotherapy. Patients with lobular carcinomas presented at a later age and had lower grade tumors that were more likely estrogen and progesterone receptor positive. Ductal carcinomas had a greater frequency of HER-2/neu amplification and increased Ki-67 rate. After chemotherapy, none of the lobular carcinomas had complete pathologic response compared with 28% of the ductal carcinomas (p = 0.01). Lobular carcinomas had more lymph node metastases. At the time of clinical follow-up, no lobular carcinomas had evidence of disease. Only one lobular carcinoma case had any histologic changes after chemotherapy compared with 37-68% of ductal carcinomas (p < 0.05). In ductal carcinomas, higher grade and negative estrogen receptor expression before chemotherapy and presence of foam cell clusters, HER 2/neu expression, and absence of lymphatic or vascular space invasion after chemotherapy correlated with pathologic response (p < 0.05). Decreased Ki-67 rate after chemotherapy correlated with survival (p = 0.024). Breast biomarker status changed in 9% of all lobular carcinomas and 19% of all ductal carcinomas. Lobular carcinomas respond poorly to neoadjuvant chemotherapy as evidence by lack of complete pathologic response and rare histologic tissue response. PMID- 19292801 TI - Identification and management of women at high risk for hereditary breast/ovarian cancer syndrome. AB - Despite advances in identifying genetic markers of high risk patients and the availability of genetic testing, it remains challenging to efficiently identify women who are at hereditary risk and to manage their care appropriately. HughesRiskApps, an open-source family history collection, risk assessment, and Clinical Decision Support (CDS) software package, was developed to address the shortcomings in our ability to identify and treat the high risk population. This system is designed for use in primary care clinics, breast centers, and cancer risk clinics to collect family history and risk information and provide the necessary CDS to increase quality of care and efficiency. This paper reports on the first implementation of HughesRiskApps in the community hospital setting. HughesRiskApps was implemented at the Newton-Wellesley Hospital. Between April 1, 2007 and March 31, 2008, 32,966 analyses were performed on 25,763 individuals. Within this population, 915 (3.6%) individuals were found to be eligible for risk assessment and possible genetic testing based on the 10% risk of mutation threshold. During the first year of implementation, physicians and patients have fully accepted the system, and 3.6% of patients assessed have been referred to risk assessment and consideration of genetic testing. These early results indicate that the number of patients identified for risk assessment has increased dramatically and that the care of these patients is more efficient and likely more effective. PMID- 19292802 TI - Locoregional relapse after breast cancer: most relapses occur late and are not clinically detected. AB - There remains controversy over follow-up after breast cancer. The National Institute for Clinical Excellence (NICE) in the United Kingdom recommends 2-3 years of follow-up for the detection of locoregional relapse. Guidelines in North America advocate much longer follow up periods. Clinicians in the UK have been reluctant to implement the NICE guidelines. Previous studies report that the rate of relapse peaks in the first 3-5 years before falling off. In this study, a retrospective analysis of rate of relapse and method of detection in 198 patients treated with conservation surgery between 1995 and 2001 has been undertaken. Median follow-up was 5.9 years. Rate of relapse is essentially constant for 10 years, with most relapses occurring after 3 years. The majority of relapse in this cohort is detected by means other than routine clinical examination, with only 16.66% of relapse detected this way. The guidelines for follow-up in the UK need revision. If follow-up is to be provided, this needs to continue for at least 10 years, if not beyond. This study casts doubt on the value of routine clinical examination. PMID- 19292803 TI - Fiberoptic ductoscopy-guided intraductal biopsy improve the diagnosis of nipple discharge. AB - Fiberoptic ductoscopy (FDS)-guided intraductal biopsy is a minimally invasive technique developed to obtain pathologic diagnoses for patients with spontaneous nipple discharge. We performed biopsies of 53 intraductal lesions from March 2006 to April 2007 followed by surgical microdochectomy. FDS-guided intraductal biopsy was shown to be a minimally invasive, safe, and convenient technique with a high ability (90.6%) to get adequate samples. Twenty-seven solitary papillomas, 12 multiple intraductal papilloma, five ductal hyperplasia, three ductal carcinoma in situ, and one invasive ductal carcinoma were diagnosed. Compared with conventional microdochectomy, FDS-guided intraductal biopsy can significantly increase the detection rate of solitary papilloma (40.7% versus 92.6%, p < 0.05). It should be a routine procedure after intraductal lesion found by screening FDS. Since it would underestimate all multiple intraductal papilloma and some (50%) cancer, microdochectomy is inevitable if biopsies show atypical ductal hyperplasia. PMID- 19292804 TI - Primary peritoneal serous carcinoma presenting as inflammatory breast cancer. AB - Metastasis to the breast from extramammary malignancies is rare. Nevertheless, its recognition is important because the prognosis and treatment differ from that of primary breast cancer. We report a unique case of primary peritoneal serous carcinoma that initially presented as inflammatory breast cancer. The patient received neoadjuvant chemotherapy for breast cancer and subsequently underwent bilateral total mastectomy and bilateral sentinel lymph node biopsy. She was found to have extensive intralymphatic carcinoma in both breasts, with only focal minimal breast parenchymal involvement, and residual metastatic carcinoma in bilateral sentinel lymph nodes. Further work-up revealed pelvic ascites and omental nodularities. The patient underwent laparoscopic bilateral salpingo oophorectomy, which revealed high-grade serous carcinoma involving both ovaries and fallopian tubes. Molecular testing of tumor from the ovary and axillary lymph node showed an identical pattern of allelic loss, confirming a common origin for both tumors. To our knowledge, this is the first reported case of an extramammary primary malignancy that not only presented as inflammatory breast cancer but also was diagnosed and initially treated as such. PMID- 19292805 TI - Breast cancer in the Middle Eastern population of California, 1988-2004. AB - This report presents the patterns of incidence, survival, and mortality of breast cancer in the Middle Eastern (ME) population of California. Cases were identified through surname recognition and population estimates were obtained from census public use files. Rates, trends, and survival in this ethnic group were compared with the non-Hispanic White (NHW) of California, as well as natives in the Middle East. Age-adjusted incidence rates for the insitu (22.8), invasive (126.2), and mortality (23.2) in ME women were significantly lower than similar rates of 26.0, 146.9, and 30.6 in the NHW women. Incidence rate in ME women in California was higher than rates in women in the Middle East. Lower rates for early stage and higher rates for late stage diagnoses in this ethnic population suggest lack of optimal access to preventive healthcare. Relative survival in the two groups is negatively associated with stage at diagnosis and is slightly higher in ME women, probably due to large numbers of lost to follow-up in ME women suggesting the presence of salmon bias. Positive association with socioeconomic standing was detected only in the NHW women. Incidence of breast cancer in ME men was significantly higher than that of NHW men. PMID- 19292806 TI - MRI of the breast: does the internet accurately report its beneficial uses and limitations? AB - As consumer use of the Internet for medical information grows, continuing evaluation of the medical content on the Internet is needed. We evaluated Internet sites describing breast magnetic resonance imaging (MRI), an emerging technology tool in breast cancer diagnosis and screening. We searched Google for sites describing breast MRI and abstracted the affiliation, content, media type, readability, and quality of 90 most popular unique sites. Over half (56%) of the sites were commercially sponsored. The content varied by site and included medical and procedural facts, information about clinical trials, grants and journal articles, as well as human interest stories. Most (82%) sites described potentially beneficial uses of breast MRI, such as further evaluation of newly diagnosed breast cancers (58%); screening women at high risk for breast cancer (54%); evaluation of abnormal breast findings (48%); screening women with dense breasts (48%) or implants (27%); and surveillance for breast cancer recurrences (24%). Approximately half (56%) of the sites described the limitations of breast MRI, most commonly false positive findings (44%) and costs (24%). Website quality, including the display of contact information, sponsorship, currency of information, authorship, and references varied. The reading level was close to high school graduate. Internet sites describing breast MRI were mostly commercially sponsored, more often described the potential beneficial uses of the procedure than its limitations, and were of variable quality and high reading level. With the lack of enforceable standards for display of medical information on the Internet, providers should encourage patients to direct their searches to the most credible sites. PMID- 19292807 TI - Enhancing the adjuvant treatment of hormone receptor positive breast cancer. AB - Aromatase inhibitors (AIs) are now regarded as the optimum hormonal therapy for postmenopausal women with hormone receptor positive breast cancer. However, it is unclear which of the currently available AIs offers patients the most effective and the best-tolerated treatment strategy. We performed a systematic review and meta-analysis of randomized-controlled trials that compared AIs (as first-line agents) with standard hormonal treatment in patients with breast cancer. The results suggest that letrozole offers a more favorable side-effect profile particularly in terms of musculoskeletal adverse events. However, the available data suggests a small survival benefit from the use of anastrozole although patients treated with anastrozole appear to have a more favorable disease profile at study entry. Examination of survival data on adjuvant tamoxifen therapy from these trials supports this observation. PMID- 19292808 TI - First report of a necrotising fasciitis of the breast following a core needle biopsy. AB - Necrotising fasciitis (NF) is the most aggressive form of soft tissue infection. We report the first case of NF of the breast following a core needle biopsy. Aggressive management including surgical debridement and vacuum therapy allowed wound healing and breast conservation. PMID- 19292809 TI - Metastatic breast cancer presenting as cancer of unknown primary associated with superior vena cava syndrome. PMID- 19292810 TI - Synchronous primary breast and ovarian cancer with ovarian cancer metastases to a breast sentinel lymph node. PMID- 19292811 TI - Solid neuroendocrine carcinoma of the breast with osteoclast-like giant cells. PMID- 19292812 TI - Primary well differentiated liposarcoma: an unusual tumor in the breast. PMID- 19292813 TI - Spider bite on a male breast: a rare case in the capital. PMID- 19292814 TI - Precipitating factors for lymphedema following surgical treatment of breast cancer: implications for patients undergoing axillary lymph node dissection. PMID- 19292815 TI - Isolated tumor cells in bone marrow and its relation with known prognostic factors in breast cancer patients. PMID- 19292816 TI - Vacuum-assisted closure dressing as an aid to salvage breast following severe postoperative infection. PMID- 19292817 TI - Travel distance to mammography and the early detection of breast cancer. PMID- 19292818 TI - Dendritic cell infiltration in lymph node positive breast carcinomas. PMID- 19292820 TI - Sensitivity to bites by the bedbug, Cimex lectularius. AB - Bedbugs are a public health problem and can cause significant economic losses, but little is known about the effects of bites on humans. We reviewed case reports and published papers on bedbug bites to assess the empirical basis of the commonly cited figure that only approximately 80% of the population are sensitive to bedbug bites. We found the sensitivity estimate to be based on only one study carried out 80 years ago. However, this study did not account for the now well established fact that only repeated exposure to external allergens leads to skin reactions. In our sample, 18 of 19 persons showed a skin reaction after bedbug exposure, but in most cases only after repeated controlled exposure. With repeated exposure, the latency between bite and skin reactions decreased from approximately 10 days to a few seconds. Our results are relevant for the hospitality industry, where apparently increasing infestation rates are likely to lead to an increase in the number of tourists and hotel employees exposed to bedbugs. Medical and public health professionals may expect to see an increase in the prevalence of people with bedbug bite sensitivity. The significance of the delayed reaction time of skin to bites may also have implications in litigation cases where people seek compensation. PMID- 19292821 TI - Effects on male fitness of removing Wolbachia infections from the mosquito Aedes albopictus. AB - Cytoplasmic incompatibility (CI) induced by maternally inherited Wolbachia bacteria is a potential tool for the suppression of insect pest species with appropriate patterns of infection. The Asian tiger mosquito Aedes albopictus (Skuse) (Diptera: Culicidae) is known to be infected by two strains of Wolbachia pipientis Hertig (Rickettsiales: Rickettsiaceae), wAlb A and wAlb B, throughout its geographical distribution. This infection pattern theoretically restricts the application of CI-based control strategies. However, Wolbachia can be horizontally transferred using embryonic microinjection to generate incompatible transfected lines harbouring a single new strain of Wolbachia. In order to assess the feasibility of this approach, the effects of Wolbachia removal on mosquito fitness need to be clearly evaluated as the removal of natural superinfection is an inescapable step of this approach. Previous research has shown that uninfected females, produced by antibiotic treatment, showed a decrease in fitness compared with those infected with Wolbachia. In this study, the effect of Wolbachia removal on male fitness was investigated. Longevity and reproductive potential (mating competitiveness and sperm capacity) were assessed in both laboratory cages and greenhouses. No differences were observed between uninfected and infected males with respect to longevity, mating rate, sperm capacity and mating competitiveness in either laboratory conditions or greenhouses. The preservation of fitness in males of Ae. albopictus deprived of natural Wolbachia infection is discussed in relation to the development of incompatible insect technique suppression strategies. Finally, the potential application of aposymbiotic males in mark-release-recapture studies is suggested. PMID- 19292822 TI - Vapour-phase activities of essential oils against antibiotic sensitive and resistant bacteria including MRSA. AB - AIMS: To determine whether essential oil (EO) vapours could reduce surface and airborne levels of bacteria including methicillin-resistant Staphylococcus aureus (MRSA). METHODS AND RESULTS: The antibacterial activity of geranium and lemongrass EO individually and blended were evaluated over a range of concentrations by direct contact and vapour diffusion. The EO were tested in vitro against a selection of antibiotic-sensitive and -resistant bacteria, including MRSA, vancomycin-resistant Enterococci (VRE), Acinetobacter baumanii and Clostridium difficile. An EO blend containing lemongrass and geranium was used to formulate BioScent that was dispersed into the environment using the ST Pro machine. The effects were variable depending on the methods used. In a sealed box environment, MRSA growth on seeded plates was reduced by 38% after 20 h exposure to BioScent vapour. In an office environment, the ST Pro machine dispersing BioScent effected an 89% reduction of airborne bacteria in 15 h, when operated at a constant output of 100%. CONCLUSIONS: EO vapours inhibited growth of antibiotic-sensitive and -resistant bacteria in vitro and reduced surface and airborne levels of bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Results suggest that EO vapours, particularly Bioscent, could be used as a method of air disinfection. PMID- 19292823 TI - Impact of two bacterial biocontrol agents on bacterial and fungal culturable groups associated with the roots of field-grown maize. AB - AIMS: To assess the impact of Bacillus amyloliquefaciens and Microbacterium oleovorans on bacterial and fungal groups associated to the roots of field-grown maize. METHODS AND RESULTS: Identification and count of bacterial and fungal culturable populations associated to the roots of maize seedlings, changes in culturable community structure according to the richness and diversity indexes concept and shifts in microbial activity through analysis of cellulolytic, ammonification and nitrification potentials were determined, in relation to kernel treatment with biological control agents. Following the treatment of maize kernels with B. amyloliquefaciens at 10(7) CFU ml(-1), an increase in bacterial diversity was observed at the rhizoplane of resultant seedlings. Bacterial richness was significantly increased at the root inner tissues of seedlings treated with Mic. oleovorans. Fusarium, Aspergillus, Penicillium and Trichoderma were the main fungal genera isolated and there population sizes were unequally affected by the addition of biocontrol agents. CONCLUSIONS: Numbers and types of isolated bacteria and fungi changed in response to the addition of biocontrol agents, while microbial activity remained unchanged with respect to control. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides an insight of the effects of proven biocontrol agents on micro-organisms naturally associated to the target crop. PMID- 19292824 TI - Functional annotation of expressed sequence tags as a tool to understand the molecular mechanism controlling flower bud development in tree peony. AB - Tree peony (Paeonia suffruticosaAndrews) is an important medicinal and ornamental plant. In China, its root bark is an important ingredient for traditional Chinese medicine. It is valued as an ornamental plant because its flower shows a wide variation in shape and color. We used flower buds at different developmental stages to construct the first cDNA library for this organism. A total of 2241 raw expressed sequence tags (ESTs) were obtained after unidirectional sequencing. After processing and assembly, they resulted in a total of 1300 unigenes [363 contigs with an average size of 3.5 ESTs (ranging from 2 up to 36) and 937 singletons]. Gene Ontology categories were assigned and further summarized into 13 broad families with biological roles according to similar functional characteristics or cellular roles. A total of 185 single nucleotide polymorphisms were detected for all contigs. We were able to detect open reading frames in the consensus sequences of 1268 unigenes. 97.5% of the ESTs showed significant similarity to sequences present in public databases. One hundred and sixty-seven short sequence repeats were obtained in the whole data set. All the analysis and information will be valuable resources for a better understanding of this important plant and also can be used for functional study in Paeoniaceae. PMID- 19292825 TI - Role of polyisoprenoids in tobacco resistance against biotic stresses. AB - Infection with avirulent pathogens, tobacco mosaic virus (TMV) or Pseudomonas syringae pv. tabaci induced accumulation of polyisoprenoid alcohols, solanesol and a family of polyprenols [from polyprenol composed of 14 isoprene units (Pren 14) to -18, with Pren-16 dominating] in the leaves of resistant tobacco plants Nicotiana tabacum cv. Samsun NN. Upon TMV infection, solanesol content was increased seven- and eight-fold in the inoculated and upper leaves, respectively, while polyprenol content was increased 2.5- and 2-fold in the inoculated and upper leaves, respectively, on the seventh day post-infection. Accumulation of polyisoprenoid alcohols was also stimulated by exogenously applied hydrogen peroxide but not by exogenous salicylic acid (SA). On the contrary, neither inoculation of the leaves of susceptible tobacco plants nor wounding of tobacco leaves caused an increase in polyisoprenoid content. Taken together, these results indicate that polyisoprenoid alcohols might be involved in plant resistance against pathogens. A putative role of accumulated polyisoprenoids in plant response to pathogen attack is discussed. Similarly, the content of plastoquinone (PQ) was increased two-fold in TMV-inoculated and upper leaves of resistant plants. Accumulation of PQ was also stimulated by hydrogen peroxide, bacteria (P. syringae) and SA. The role of PQ in antioxidant defense in cellular membranous compartments is discussed in the context of the enzymatic antioxidant machinery activated in tobacco leaves subjected to viral infection. Elevated activity of several antioxidant enzymes (ascorbate peroxidase, guaiacol peroxidase, glutathione reductase and superoxide dismutase, especially the CuZn superoxide dismutase isoform) and high, but transient elevation of catalase was found in inoculated leaves of resistant tobacco plants but not in susceptible plants. PMID- 19292826 TI - Extreme host plant conservatism during at least 20 million years of host plant pursuit by oak gallwasps. AB - Diversification of insect herbivores is often associated with coevolution between plant toxins and insect countermeasures, resulting in a specificity that restricts host plant shifts. Gall inducers, however, bypass plant toxins and the factors influencing host plant associations in these specialized herbivores remain unclear. We reconstructed the evolution of host plant associations in Western Palaearctic oak gallwasps (Cynipidae: Cynipini), a species-rich lineage of specialist herbivores on oak (Quercus). (1) Bayesian analyses of sequence data for three genes revealed extreme host plant conservatism, with inferred shifts between major oak lineages (sections Cerris and Quercus) closely matching the minimum required to explain observed diversity. It thus appears that the coevolutionary demands of gall induction constrain host plant shifts, both in cases of mutualism (e.g., fig wasps, yucca moths) and parasitism (oak gallwasps). (2) Shifts between oak sections occurred independently in sexual and asexual generations of the gallwasp lifecycle, implying that these can evolve independently. (3) Western Palaearctic gallwasps associated with sections Cerris and Quercus diverged at least 20 million years ago (mya), prior to the arrival of oaks in the Western Palaearctic from Asia 5-7 mya. This implies an Asian origin for Western Palaearctic gallwasps, with independent westwards range expansion by multiple lineages. PMID- 19292828 TI - Current issues in neuroimaging. PMID- 19292827 TI - Imaging cerebral blood flow in the cognitively normal aging brain with arterial spin labeling: implications for imaging of neurodegenerative disease. AB - INTRODUCTION: Arterial spin labeling (ASL) is a safe, noninvasive imaging method for evaluating cerebral blood flow (rCBF). The purpose of this article is to present ASL imaging features of 38 elderly cognitively normals (CN) with their rCBF values and an averaged profile of targeted anatomic regions rCBF values. METHODS: Thirty-eight CN underwent MR imaging especially ASL with voxel morphometric techniques fusing the MR anatomical and ASL images to a standard reference brain (colin27). The ASL images were fused to echo planar images, which were then coregistered to high-resolution anatomical SPGR images. rCBF was calculated per region of interest using a modified continuous arterial spin labeling (CASL) convolutional method. Anatomical regions were selected and identified by the Talairach atlas in SPM2. RESULTS: We identified areas of decreased and increased perfusion (compared to the averaged rCBF of all 38 CN) corresponding to decreased and increased quantified rCBF. The most common sites for decreased perfusion were precuneus (53%), superior temporal (48%), and orbitofrontal (37%), and for increased perfusion the caudate (39%), posterior cingulate (34%), anterior cingulate (32%), and amygdala (32%). CONCLUSION: There are regional variations in rCBF both increased and decreased with the posterior cingulate and precuneus cortex showing the highest averaged values and signal intensity (bright spots). These variations represent the normal profile of a CN elderly brain, with higher perfusion in areas associated with cognition, memory, and behavior. It is necessary to understand these normal variations in order to determine if there are perfusion changes in ASL detected in neurodegenerative disorders such as Alzheimer's disease. PMID- 19292829 TI - Custom-devised and generic digital enhancement of images for people with maculopathy. AB - AIM: The purpose of this study was to compare the effectivity, in terms of the potential usefulness, of digital filters based on either contrast sensitivity (CS) or supra-threshold contrast matching (CM) in enhancing pictures images for people with maculopathy and to investigate whether generic filters (not based on an individual's vision loss) are equally as effective. Effectivity is measured by changes in perceived visibility. METHODS: Thirty-five subjects with maculopathy, aged 20-92 years, took part [13 atrophic age-related macular degeneration (ARMD), 14 exudative ARMD, and 8 juvenile macular dystrophy (JMD)]. CS and supra threshold CM were measured. A range of CS filters (1 or 2-octave wide band-pass filter using a Gabor or polynomial envelope) with different strengths were developed based on the ratio of the individual's contrast threshold and that of a normal age-related group. Similarly filters were developed based on CM at 3.6% and 27.9% contrast. The following generic filters were also applied with different 'strengths': edge enhancement; sharpening; contrast enhancement; Peli's adaptive enhancement; difference of Gaussian; and an equi-emphasis band-pass filter. The filters were applied to images of faces and general scenes. Subjects were asked to rank the perceived visibility of images (to obtain the best version of each filter) and then to rate the perceived visibility of each image filtered with a particular filter. RESULTS: In general, subjects with atrophic ARMD and JMD preferred the weaker versions of most of the filters, while those with exudative ARMD did not show such a clear preference. Generally, images of faces were preferred with less enhancement than scenes. The filters based on CM were rated as giving significant improvement, while those based on CS and peak emphasis were not preferred. Of the generic filters, the Peli adaptive enhancement filter was most frequently rated as giving a significant improvement (p < 0.05) followed by the contrast enhancement filter. They gave the same perceived enhancement as the custom-devised filters. CONCLUSIONS: Generic filters, which are easier to apply than the custom-devised filters, are appropriate for rehabilitation purposes. PMID- 19292830 TI - Correctable and non-correctable visual impairment among young males: a 12-year prevalence study of the Military Service in Poland. AB - PURPOSE: To evaluate the prevalence of correctable and non-correctable visual impairment in a representative sample of young males commissioned for Military Service in Poland. MATERIAL AND METHODS: Data concerning vision status was retrospectively reviewed in 969 subjects of European Caucasian origin, most of whom live and have lived in Poland. They were selected from the original database comprising 105017 subjects examined in the period 1993-2004. Based on the age of subjects they were divided into two groups; group I aged 18-24 and group II aged 25-34 years. Visual impairment was defined as distance visual acuity of <20/40 in one or both eyes. Non-correctable impairment was defined as that which was not eliminated by refractive correction. RESULTS: A total of 1938 eyes of 969 white males were examined. There was statistically significant association between rates of visual impairment and increasing age (p < 0.001). Visual impairment was found in 128 (13.2%) subjects in at least one eye. Non-correctable visual impairment was found in 12 (1.2%) subjects. Amblyopia was the main cause, accounting for eight cases (66.67%). Correctable visual impairment was found in the remaining 116 (12.0%) patients. Among them myopia was the most common refractive error and accounted for 75.8%. Differences between age-specific rates of refractive errors were statistically significant (p = 0.003). CONCLUSIONS: Appropriate refractive correction improves visual acuity in most subjects presenting with visual impairment. There was a relatively low prevalence of non correctable visual impairment in a population of young adults in Poland, and this was mainly due to amblyopia. PMID- 19292831 TI - Blockade of K(ATP) channels reduces endothelial hyperpolarization and leukocyte recruitment upon reperfusion after hypoxia. AB - Ischemia/reperfusion injury in renal transplantation leads to slow or initial nonfunction, and predisposes to acute and chronic rejection. In fact, severe ischemia reperfusion injury can significantly reduce graft survival, even with modern immunosuppressive agents. One of the mechanisms by which ischemia/reperfusion causes injury is activation of endothelial cells resulting in inflammation. Although several therapies can be used to prevent leukocyte recruitment to ischemic vessels (e.g. antiadhesion molecule antibodies), there have been no clinical treatments reported that can prevent initial immediate neutrophil recruitment upon reperfusion. Using intravital microscopy, we describe abrogation of immediate neutrophil recruitment to ischemic microvessels by the K(ATP) antagonist glibenclamide (Glyburide). Further, we show that glibenclamide can reduce leukocyte recruitment in vitro under physiologic flow conditions. ATP regulated potassium channels (K(ATP)) are important in the control of cell membrane polarization. Here we describe profound hyperpolarization of endothelial cells during hypoxia, and the reduction of this hyperpolarization using glibenclamide. These findings suggest that control of endothelial membrane potential during ischemia may be an important therapeutic tool in avoiding ischemia/reperfusion injury, and therefore, enhancing transplant long-term function. PMID- 19292832 TI - Influence of insulin resistance on hepatic fibrosis and steatosis in hepatitis C virus (HCV) mono-infected compared with HIV-HCV co-infected patients. AB - BACKGROUND: Insulin resistance (IR), the major feature of the metabolic syndrome, is also common in patients with chronic HCV infection. Liver fibrosis and steatosis are known potential outcome of chronic hepatitis B or C infection. Studies have shown that HIV positive individuals co-infected with HCV have more rapid live disease progression than those with HIV alone. Few data have reported the influence of IR on steatosis and fibrosis in the context of HIV-HCV coinfection. AIM: To test the association among insulin resistance (IR), liver fibrosis and liver steatosis in HIV-HCV and HCV-infected patients. PATIENTS AND METHODS: A total of 170 HIV-HCV-infected patients matched by age, gender and genotype with 170 HCV mono-infected patients were included. Patients were considered to be IR when the homeostasis model assessment of IR >2. Significant fibrosis was considered if METAVIR >or=F2 and significant steatosis if >or=10%. RESULTS: Insulin resistance was independently associated in HCV patients with fibrosis [odds ratio (OR) = 2.04 (95% CI 1.02-4)], a body mass index (BMI) >25 kg/m(2) [OR = 3.33 (1.47-7.69)] and steatosis [OR = 3.33 (1.67-6.67)]. Fibrosis >or=F2 was associated in HCV patients with high liver activity grade (>or=A2) [OR = 8.33 (3.85-16.67)], male gender [OR = 3.03 (1.33-7.14)] and IR [OR = 2.44 (1.15 5)]. In HIV-HCV patients, >or=A2 [OR = 5.56 (1.64-20)] was associated with fibrosis. Steatosis >or=10% was associated in HCV patients with IR [OR = 3.13 (1.59-6.25) and >or=F2 (OR = 2.22 (1.15-4.17)]. In HIV-HCV, a BMI >25 kg/m(2) [OR = 3.85 (1.64-9.10)], >or=A2 [OR = 2.16 (1.02-4.55); P = 0.044] and nucleoside reverse transcriptase inhibitor [OR = 3.61 (1.19-10.96); P = 0.023] were independently associated with significant liver steatosis. CONCLUSIONS: Insulin resistance is associated with liver fibrosis and steatosis in HCV mono-infected, but not in HIV-HCV co-infected patients. Significant liver fibrosis is associated with IR independent of liver steatosis only in HCV mono-infected patients. PMID- 19292833 TI - Genetic and environmental influences on serum lipids and the effects of puberty: a Chinese twin study. AB - AIM: To study the contribution of genes and environment on the variation of serum lipids and the effects of puberty. METHODS: In total, 314 same-sex twin pairs aged 5-18 years were studied. Puberty was marked physiologically by spermarche/menarche, and model fitting was used to analyse the genetic and environmental variance and its difference before and after puberty. RESULTS: Lipid levels were different before and after puberty. The genetic factor had an important influence on lipid levels; the heritability estimates of total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), lipoprotein(a) (Lp(a)) and apolipoprotein E (ApoE) were between 49 and 86%. The total phenotypic variances of TC, HDL, LDL, Lp(a) and ApoE decreased after puberty, mainly as a result of decrease of genetic variance, even though the common environmental variance for HDL, Lp(a) and ApoE increased. CONCLUSION: Genes and the environment have different effects on the levels of different lipids. The shared environmental effects on lipids are very important in children. The role of puberty on lipids deserves future study. PMID- 19292834 TI - Core competencies for hair restoration surgeons recommended by the International Society of Hair Restoration Surgery. AB - BACKGROUND: Because hair restoration surgery has changed so significantly, the International Society of Hair Restoration Surgery (ISHRS) recently developed and published a Core Curriculum for Hair Restoration Surgery (CCHRS). The ISHRS organized a task force to develop training programs that would not only present the CCHRS but also provide the practical experience necessary to allow a physician to practice safe, aesthetically sound hair restoration surgery. The task force recognized early on that identification of core competencies for hair restoration surgeons was essential to guiding the development of these training experiences. This article presents the competencies that have been identified. OBJECTIVE: The intent of the Core Competencies for Hair Restoration Surgery is to outline the knowledge and skills that are essential to accurately diagnose and treat hair loss, to ensure patient safety, and to optimize aesthetic results. The ISHRS hopes that all existing surgery and dermatology training programs teaching hair restoration surgery procedures will find the Core Competencies useful in developing their curriculums. METHODS: The Core Competencies were developed through an organized review of the CCHRS by a team of experienced hair restoration surgeons and educators and reviewed and approved by the ISHRS Board of Governors. RESULTS: The diversity of these competencies demonstrate that contemporary hair restoration surgery is a specialty requiring knowledge of several medical disciplines, including genetics, endocrinology, dermatology, tissue preservation, and surgery. CONCLUSION: The International Society of Hair Restoration Surgery believes identification of these Core Competencies is an important contribution to physician education in hair restoration surgery, and physicians who demonstrate competency in these skills will satisfy patients with contemporary results in a safe environment. PMID- 19292835 TI - Time in motion--testing efficiency in the dermatology procedure setting. AB - BACKGROUND: Time-in-motion studies are validated means of determining efficiency and identifying sources of delay in a complex working environment. This analysis technique was applied to the dermatological surgical services provided by a university hospital department of dermatology. OBJECTIVE: To identify any sources of low efficiency in dermatological surgical service provision and to make recommendations for increasing efficiency. METHOD: Three types of dermatology surgical sessions were observed (consultant-led procedure list, nurse-led procedure list, and dermatology day-surgery list). Activities of staff were noted every 2 minutes throughout the sessions, and the data were compiled for processing in Microsoft Excel. RESULTS: The results show that treatment of dermatology patients with day surgery was less efficient and cost effective than the other settings. Consultant-led procedure lists were the most efficient. Nurse led sessions were less expensive but also less efficient than the consultant-led lists. Paperwork and generic communication were identified as major constituents of non-operating time. CONCLUSIONS: The most efficient setting for dermatological procedures is the consultant-led procedure list. Paperwork and generic communication are identified as sources of service delay. Suggestions are made regarding possible solutions to these problems. We demonstrate the advantages of broad, objective appraisal of practice as opposed to numerical target-driven approaches that fail to address specific aspects of efficiency. PMID- 19292836 TI - Effects of orlistat on visceral fat after liposuction. AB - BACKGROUND: Liposuction can aggravate metabolic complications associated with obesity. It has been shown that the recovery of weight lost through these interventions is associated with body fat redistribution toward the visceral cavity, increasing metabolic risk factors for coronary heart disease such as insulin resistance and high triglyceride levels. OBJECTIVES: The aim of this study was to evaluate the consequences of liposuction on body mass redistribution and metabolic parameters 6 months after surgery and to evaluate the use of orlistat treatment (tetrahydrolipstatin) in controlling these parameters. METHODS: A population of 31 women with a mean body mass index of 26.17+/-3.9 kg/m(2) and undergoing liposuction of more than 1,000 cm(3), was studied. Twelve of them were treated postsurgery with 120 mg of orlistat every 8 hours for the following 6 months. Anthropometric, analytical, and radiological (computed tomography) tests were performed to quantify visceral fat area before surgery and 6 months after surgery. RESULTS: Despite weight loss after liposuction, visceral fat was not modified. Patients treated with orlistat showed a greater reduction in visceral fat, although not statistically significant. Orlistat use induced a reduction in low-density lipoprotein cholesterol values of 20.0+/-22.5 mg/dL, compared with an increase of 8.46+/-20.1 mg/dL in controls (p=.07). CONCLUSIONS: Visceral fat does not decrease despite weight loss after liposuction. Orlistat use postliposuction might be a useful tool because it shows a tendency to reduce visceral fat and improve blood lipids profile. PMID- 19292837 TI - Effect of a novel low-energy pulsed-light device for home-use hair removal. AB - BACKGROUND: Removal of unwanted hair is the most popular skin treatment worldwide. Over the past decade, various lasers and light sources for epilation have been advocated for use in an office setting, although most people continue to treat unwanted hair with a variety of temporary physical methods (e.g., waxing, shaving) in a home setting, presumably due to cost and convenience factors. OBJECTIVES: To evaluate the safety and efficacy of a low-energy pulsed light device intended for home-use hair removal. MATERIALS AND METHODS: Twenty women (skin phototypes I-IV) with dark terminal hair in nonfacial sites (axilla, forearms, inguinal region, legs) self-administered three treatments at 2-week intervals using a handheld intense-pulsed-light device. Matched untreated skin sites were also studied. Hair counts and clinical photographs were obtained pretreatment and at 1, 3, and 6 months after the third treatment. Side effects and patient satisfaction scores were recorded. RESULTS: All patients showed a positive clinical response to treatment, with reduction of unwanted hair. No reduction of hair was noted in untreated matched areas. Hair counts were reduced 37.8% to 53.6% 6 months after the three treatments. Skin region influenced clinical response, with lower legs exhibiting greater hair reduction than arms and inguinal and axillary areas. Mild erythema was experienced in 25% of patients, but no other side effects or complications were encountered. Patient satisfaction scores were high, with all patients stating that they would purchase the device for future home use. CONCLUSIONS Low-energy pulsed light can be applied safely and effectively for at-home hair removal in a variety of nonfacial locations and skin phototypes I-IV. PMID- 19292838 TI - Eyebrow transplantation in Asians. PMID- 19292839 TI - Eruptive keratoacanthomas on the legs after fractional photothermolysis: report of two cases. PMID- 19292840 TI - Eccrine syringofibroadenoma-like change adjacent to a squamous cell carcinoma: potential histologic pitfall in Mohs micrographic surgery. PMID- 19292841 TI - Generalized chrysiasis improved with pulsed dye laser. PMID- 19292842 TI - Rhinophyma treated with cryosurgery. PMID- 19292843 TI - Surgical treatment of aplasia cutis congenita with acellular dermal graft and cultured epithelial autograft. PMID- 19292844 TI - Three rhombic flaps for repair of an upper lip wound. PMID- 19292846 TI - Otoacoustic emissions: a valid, efficient first-line hearing screen for preschool children. AB - BACKGROUND: Normal hearing during the preschool years is essential for speech, language, social, emotional, and preacademic development. Children of low socioeconomic status may be particularly vulnerable to the negative effects of late identification and intervention. While a mass-screening effort focused on preschool children does not have broad support, focused screening remains important to identify children at risk. This project was conducted to address 3 primary aims: develop and implement an initial hearing screen using transient evoked otoacoustic emissions (TEOAEs) for at-risk preschoolers, verify speed and tolerability of the screen, and assess the test performance of TEOAEs screening compared to pure tone audiometry in a group of 142 preschool children. METHODS: A total of 744 preschool children attending preschools in an underserved, urban community completed TEOAEs screening by a school nurse. A secondary cohort of 142 children was screened first by TEOAEs and then followed by pure tone audiometry and results compared. RESULTS: A total of 680 children passed screening. Forty one children (5.5%) had a "refer" result. Two-year-olds had the highest refusal rate (10.5%). Mean testing time was 43 seconds per ear. Secondary cohort analysis revealed 1 subject did not pass either TEOAEs or pure tone screening; no subject passed TEOAEs and then did not pass pure tone audiometry. TEOAEs screening test sensitivity was 1.00 (95% confidence interval 0.054-1.00) and specificity 0.94 (0.88-0.97). CONCLUSIONS: TEOAEs screening performed by school nurses is a fast, efficient, and feasible model. Children who pass TEOAEs screening have a very high likelihood of being free from hearing impairment. Application may be particularly relevant in underserved communities. PMID- 19292847 TI - Relation of school environment and policy to adolescent physical activity. AB - BACKGROUND: Physical activity (PA) declines as children and adolescents age. The purpose of this study was to examine how specific school factors relate to youth PA, TV viewing, and body mass index (BMI). METHODS: A sample of 12- to 18-year old adolescents in 3 cities (N = 165, 53% females, mean age 14.6 +/- 1.7 years, 44% nonwhite) completed surveys assessing days of physical education (PE) class per week, school equipment accessibility, after-school supervised PA, and after school field access. Regression analyses were conducted to examine relationships between these school factors and PA at school facilities open to the public (never active vs active), overall PA level (days per week physically active for 60 minutes), BMI z score, and TV watching (hours per week). RESULTS: Adjusting for demographics, days of PE per week and access to school fields after school were correlated with overall PA (beta= 0.286, p = .002, semipartial correlation .236 and beta= 0.801, p = .016, semipartial correlation .186, respectively). The association between after-school field access and overall PA was mediated by use of publicly accessible school facilities for PA. After-school supervised PA and school PA equipment were not associated with overall PA. In adjusted regression analyses including all school factors, days of PE remained correlated to overall PA independent of other school factors (beta= 0.264, p = .007, semipartial correlation = .136). There were no associations between school factors and BMI or TV watching. CONCLUSIONS: Based on these study findings, PE is a promising intervention to address improving overall adolescent PA within the school setting. PMID- 19292848 TI - Social environments and physical aggression among 21,107 students in the United States and Canada. AB - BACKGROUND: Physical aggression is an important issue in North American populations. The importance of students' social environments in the occurrence of physical aggression requires focused study. In this study, reports of physical aggression were examined in relation to social environment factors among national samples of students from Canada and the United States. METHODS: Students in grades 6-10 from the United States (n = 14,049) and Canada (n = 7058) who had participated in the Health Behaviour in School-aged Children Survey (HBSC) were studied. Rates of students' physical aggression were compared between the 2 countries. School, family, socioeconomic, and peer-related factors were considered as potential risk factors. A simple social environment risk score was developed using the US data and was subsequently tested in the Canadian sample. RESULTS: Risks for physical aggression were consistently higher among United States versus Canadian students, but the magnitude of these differences was modest. The relative odds of physical aggression increased with reported environmental risk. To illustrate, US boys in grades 6-8 reporting the highest social environment risk score (5+) experienced a relative odds of physical aggression 4.02 (95% CI 2.7-5.9) times higher than those reporting the lowest score (adjusted OR for risk scores 0 through 5+ was 1.00, 1.19, 2.10, 2.01, 3.71, and 4.02, respectively, p(trend) < .001). CONCLUSIONS: Unexpectedly, rates of physical aggression and associations between social environments and students' aggression were remarkably similar in Canada and the United States. Family, peer, and school social environments serve as risk or protective factors, with significant cumulative impact on physical aggression in both countries. Given the observed high rates and the many negative effects of aggression on long-term health, school policies aimed at the reduction of such behavior remain a clear priority. PMID- 19292849 TI - Trends and subgroup differences in transportation-related injury risk and safety behaviors among US high school students, 1991-2007. AB - BACKGROUND: Seventy percent of unintentional injury-related fatalities--the leading cause of death among youth in the United States--are motor vehicle traffic related. Examining traffic-related safety, therefore, is crucial to public health. This study examines trends in traffic safety issues among US high school students: helmet use while riding a bicycle, seat belt use as a passenger, driving when drinking alcohol, and riding in a car with a driver who had been drinking alcohol. METHODS: Data from the 1991-2007 national Youth Risk Behavior Surveys (YRBS) were analyzed. The YRBS is a self-administered, anonymous survey that uses a national probability sample of US students in public and private schools in grades 9-12. Demographic subgroup differences were determined for 2007 data using t tests. Temporal changes were analyzed using logistic regression analyses. RESULTS: From 1991 to 2007, the percentage of high school students who rarely or never wore bicycle helmets decreased from 96.2% to 85.1%; decreases were also seen in the percentage who never wore a seat belt (from 25.9% to 11.1%), rode with a driver who had been drinking alcohol (39.9-29.1%), and who drove when drinking alcohol (16.7-10.5%). CONCLUSIONS: Although the trends are encouraging, many students still put themselves at risk. Policy approaches (eg, state or local laws or ordinances) complemented by community and school programs may be the best approach to reducing transportation-related injuries and fatalities. PMID- 19292850 TI - Responses to positive results from suspicionless random drug tests in US public school districts. AB - BACKGROUND: Little is known about the context in which school-based suspicionless random drug testing (SRDT) occurs. The primary purpose of the current study was to describe school districts' responses to students' first positive result in districts with SRDT programs. METHODS: Data were collected in spring 2005 from 1612 drug prevention coordinators in a nationally representative sample of 1922 school districts (83.9% response rate), of which 205 districts reported SRDT in high school grades. RESULTS: Respondents reported an array of consequences for students with an initial positive SRDT, including requiring parents or guardians to meet with school officials (88.4%) and requiring students to participate in an education, counseling, or treatment program (60.8%). However, some districts also reported consequences contraindicated by federal advisory guides, such as notifying law enforcement officials (45.1%) and suspending the student from an athletic team (65.0%) or from school (31.0%). Some respondents may have conflated their districts' responses to for cause and random tests. Districts generally had available key services for students testing positive, including professional counseling for substance use problems (87.3%) and referrals to counseling services (91.9%). CONCLUSIONS: More understanding is needed of schools' responses to students who test positive following the administration of SRDT, available advisory guides concerning best practices should be more effectively disseminated, and appropriate training and technical assistance should be available to schools with SRDTs. PMID- 19292851 TI - Promoting tobacco-free school policies through a statewide media campaign. AB - BACKGROUND: Comprehensive, enforced tobacco-free school (TFS) policies lead to significant reductions in youth tobacco use. North Carolina is the first state in the United States to develop a statewide mass media campaign to promote the adoption of and compliance with TFS policies. METHODS: In order to guide campaign development, researchers conducted a literature review as well as interviews with 45 TFS-policy experts, stakeholders, and North Carolina legislators. The experts included state and national TFS researchers and advocates, and the majority of stakeholders were North Carolina school administrators and personnel selected because of their personal knowledge of and experience with TFS policy. Interviewees provided information on messages they believed would be most salient to highlight in the media campaign and the best type of people to appear in ads. RESULTS: Recommended themes included (1) a positive message about TFS becoming the norm in the state, (2) experiences of school districts that had successfully passed TFS policies, (3) the importance of adult role modeling, and (4) personal stories from youth about the importance of TFS policy. Recommended people to appear in ads included youth and adults with a personal connection to and experience with TFS policy. Using these recommended themes and people, the TFS media campaign began in the fall of 2006. In the 8 months following the campaign launch, 9 additional school districts adopted TFS policies, increasing the total from 78 to 87 (out of 115) by June 2007. In July 2007, the North Carolina legislature passed legislation mandating that all school districts adopt TFS policies by August 2008. CONCLUSIONS: Media campaigns can serve as part of a comprehensive strategy to advance TFS policies. Other states should consider these results in designing and evaluating a media campaign to promote adoption of and compliance with TFS policies. PMID- 19292853 TI - "What would you do, what if it's you?" Strategies to deal with a bully. PMID- 19292852 TI - Patterns of vaginal, oral, and anal sexual intercourse in an urban seventh-grade population. AB - BACKGROUND: This study examines the prevalence of vaginal, oral, and anal intercourse among a population of urban, public middle school students, the characteristics of early sexual initiators, and the sequence of sexual initiation. Such data are limited for early adolescents. METHODS: A total of 1279 seventh-grade students (57.3% female, 43.6% black, and 41.8% Hispanic), mean age 12.5 years (SD = 0.63) from 10 middle schools in a large southeastern US public school district completed a cross-sectional survey using audio computer-assisted self-interviews. Main outcomes included lifetime and past 3-months' experience of vaginal, oral, and anal sex; condom use; age of initiation; and number of lifetime partners. RESULTS: Overall, 12.0% of students had engaged in vaginal sex, 7.9% in oral sex, and 6.5% in anal sex. Among students who had initiated intercourse, approximately two thirds were currently sexually active and one quarter reported > or =4 lifetime partners. Six percent had engaged in 1 type of intercourse, 4% in 2 types of intercourse, and 4% in all 3 types of intercourse; vaginal sex was typically initiated at an earlier or at the same age as other types of intercourse. Only 2% had engaged in oral sex without engaging in vaginal sex. Although black students were significantly more likely to have initiated sex compared to other racial/ethnic groups, Hispanic students who had initiated each type of intercourse were more likely to be currently sexually active and to have engaged in recent unprotected sex. CONCLUSIONS: A small percentage of early adolescents are engaging in multiple sexual behaviors. These findings have implications for early adolescent school-based sexual health education. PMID- 19292855 TI - Immunomagnetic isolation of CD4+CD25+FoxP3+ natural T regulatory lymphocytes for clinical applications. AB - Although CD4(+)/CD25(+) T regulatory cells (T(regs)) are a potentially powerful tool in bone marrow transplantation, a prerequisite for clinical use is a cell separation strategy complying with good manufacturing practice guidelines. We isolated T(regs) from standard leukapheresis products using double-negative selection (anti-CD8 and anti-CD19 monoclonal antibodies) followed by positive selection (anti-CD25 monoclonal antibody). The final cell fraction (CD4(+)/CD25(+)) showed a mean purity of 93.6% +/- 1.1. Recovery efficiency was 81.52% +/- 7.4. The CD4(+)/CD25(+bright) cells were 28.4% +/- 6.8. The CD4(+)/CD25(+) fraction contained a mean of 51.9% +/- 15.1 FoxP3 cells and a mean of 18.9% +/- 11.5 CD127 cells. Increased FoxP3 and depleted CD127 mRNAs in CD4(+)CD25(+)FoxP3(+) cells were in line with flow cytometric results. In Vbeta spectratyping the complexity scores of CD4(+)/CD25(+) cells and CD4(+)/CD25(-) cells were not significantly different, indicating that T(regs) had a broad T cell receptor repertoire. The inhibition assay showed that CD4(+)/CD25(+) cells inhibited CD4(+)/CD25(-) cells in a dose-dependent manner (mean inhibition percentages: 72.4 +/- 8.9 [ratio of T responder (T(resp)) to T(regs), 1:2]; 60.8% +/- 20.5 (ratio of T(resp) to T(regs), 1:1); 25.6 +/- 19.6 (ratio of T(resp) to T(regs), 1:0.1)). Our study shows that negative/positive T(reg) selection, performed using the CliniMACS device and reagents, enriches significantly CD4(+)CD25(+)FoxP3(+) cells endowed with immunosuppressive capacities. The CD4(+)CD25(+)FoxP3(+) population is a source of natural T(reg) cells that are depleted of CD8(+) and CD4(+)/CD25(-) reacting clones which are potentially responsible for triggering graft-versus-host disease (GvHD). Cells isolated by means of this approach might be used in allogeneic haematopoietic cell transplantation to facilitate engraftment and reduce the incidence and severity of GvHD without abrogating the potential graft-versus-tumour effect. PMID- 19292856 TI - Who's the faster runner: the cheetah or the rabbit? PMID- 19292857 TI - An in vivo confocal microscopic analysis of Salzmann's nodular degeneration: pre- and post-surgical intervention. PMID- 19292858 TI - Macular hole following intravitreal ranibizumab injection for choroidal neovascular membrane caused by age-related macular degeneration. PMID- 19292859 TI - Argon laser trabeculoplasty for late glaucoma after intravitreal triamcinolone. PMID- 19292860 TI - New perspectives on folate transport in relation to alcoholism-induced folate malabsorption--association with epigenome stability and cancer development. AB - Folates are members of the B-class of vitamins, which are required for the synthesis of purines and pyrimidines, and for the methylation of essential biological substances, including phospholipids, DNA, and neurotransmitters. Folates cannot be synthesized de novo by mammals; hence, an efficient intestinal absorption process is required. Intestinal folate transport is carrier-mediated, pH-dependent and electroneutral, with similar affinity for oxidized and reduced folic acid derivatives. The various transporters, i.e. reduced folate carrier, proton-coupled folate transporter, folate-binding protein, and organic anion transporters, are involved in the folate transport process in various tissues. Any impairment in uptake of folate can lead to a state of folate deficiency, the most prevalent vitamin deficiency in world, affecting 10% of the population in the USA. Such impairments in folate transport occur in a variety of conditions, including chronic use of ethanol, some inborn hereditary disorders, and certain diseases. Among these, ethanol ingestion has been the major contributor to folate deficiency. Ethanol-associated folate deficiency can develop because of dietary inadequacy, intestinal malabsorption, altered hepatobiliary metabolism, enhanced colonic metabolism, and increased renal excretion. Ethanol reduces the intestinal and renal uptake of folate by altering the binding and transport kinetics of folate transport systems. Also, ethanol reduces the expression of folate transporters in both intestine and kidney, and this might be a contributing factor for folate malabsorption, leading to folate deficiency. The maintenance of intracellular folate homeostasis is essential for the one-carbon transfer reactions necessary for DNA synthesis and biological methylation reactions. DNA methylation is an important epigenetic determinant in gene expression, in the maintenance of DNA integrity and stability, in chromosomal modifications, and in the development of mutations. Ethanol, a toxin that is consumed regularly, has been found to affect the methylation of DNA. In addition to its effect on DNA methylation due to folate deficiency, ethanol could directly exert its effect through its interaction with one-carbon metabolism, impairment of methyl group synthesis, and affecting the enzymes regulating the synthesis of S adenosylmethionine, the primary methyl group donor for most biological methylation reactions. Thus, ethanol plays an important role in the pathogenesis of several diseases through its potential ability to modulate the methylation of biological molecules. This review discusses the underlying mechanism of folate malabsorption in alcoholism, the mechanism of methylation-associated silencing of genes, and how the interaction between ethanol and folate deficiency affects the methylation of genes, thereby modulating epigenome stability and the risk of cancer. PMID- 19292861 TI - Solubility-dependent structural formation of a 25-residue, natively unfolded protein, induced by addition of a seven-residue peptide fragment. AB - To elucidate the architectural principle of protein structure, we focused on sequestration from solvent, which is a common characteristic of folding and self associative precipitation. Because protein solubility can be regarded as a basis for the potential ability to sequester from solvent, we assume that poorly soluble proteins tend not only to precipitate, but also to form solution structures. To examine this, the solubility of a 25-residue, natively unfolded protein, modified from a zinc-finger domain of transcription factor Sp1, was disturbed by adding a seven-residue hydrophobic peptide fragment to the C terminus. NMR and ultracentrifuge measurements of the resulting sequence showed that a dissolved species forms an alpha-helical structure in a 15-20 molecule oligomer. To elucidate the mechanism by which the structure forms, we prepared two variants in which the added fragments are less hydrophobic; the structural stabilities were then measured at various pH values. A fairly good correlation was observed between stability and hydration potential, whereas a much stronger correlation was observed between stability and solubility, indicating that the stability is more strongly dependent on the ability to precipitate than on dehydration. These results show that, among poorly soluble protein molecules, dissolved species can be transformed from the solvent-exposed unfolded state into a loosely packed structure via intermolecular interactions. Because decreasing the protein solubility does not require the primary sequence to have a sophisticated design, such a protein structure might form readily and frequently, compared with the well-packed structure found in native proteins. PMID- 19292862 TI - The in vitro nuclear aggregates of polyamines. AB - Natural polyamines (putrescine, spermidine, and spermine) self-assemble in a simulated physiological environment (50 mm sodium phosphate buffer, pH 7.2), generating in vitro nuclear aggregates of polyamines (ivNAPs). These supramolecular compounds are similar in structure and molecular mass to naturally occurring cellular nuclear aggregates of polyamines, and they share the ability of NAPs to interact with and protect the genomic DNA against nuclease degradation. Three main ivNAP compounds were separated by gel permeation chromatography. Their elution was carried out with 50 mm sodium phosphate buffer supplemented with 150 mm NaCl. Freezing and thawing of selected chromatographic fractions obtained by GPC runs in which the mobile phase was sodium phosphate buffer not supplemented with NaCl yielded three different microcrystallites, specifically corresponding to the ivNAPs, all of which were able to bind DNA. In this study, we demonstrated that in vitro self-assembly of polyamines and phosphates is a spontaneous, reproducible and inexpensive event, and provided the indications for the production of the ivNAPs as a new tool for manipulating the genomic DNA machinery. PMID- 19292863 TI - The hemoglobins of the sub-Antarctic fish Cottoperca gobio, a phyletically basal species--oxygen-binding equilibria, kinetics and molecular dynamics. AB - The dominant perciform suborder Notothenioidei is an excellent study group for assessing the evolution and functional importance of biochemical adaptations to temperature. The availability of notothenioid taxa in a wide range of latitudes (Antarctic and non-Antarctic) provides a tool to enable identification of physiological and biochemical characteristics gained and lost during evolutionary history. Non-Antarctic notothenioids belonging to the most basal families are a crucial source for understanding the evolution of hemoglobin in high-Antarctic cold-adapted fish. This paper focuses on the structure, function and evolution of the oxygen-transport system of Cottoperca gobio, a sub-Antarctic notothenioid fish of the family Bovichtidae, probably derived from ancestral species that evolved in the Antarctic region and later migrated to lower latitudes. Unlike most high-Antarctic notothenioids, but similar to many other acanthomorph teleosts, C. gobio has two major hemoglobins having the beta chain in common. The oxygen-binding equilibria and kinetics of the two hemoglobins have been measured. Hb1 and Hb2 show strong modulation of oxygen-binding equilibria and kinetics by heterotropic effectors, with marked Bohr and Root effects. In Hb1 and Hb2, oxygen affinity and subunit cooperativity are slightly higher than in most high Antarctic notothenioid hemoglobins. Hb1 and Hb2 show similar rebinding rates, but also show significant dynamic differences that are likely to have functional consequences. Molecular dynamic simulations of C. gobio Hb1 were performed on the dimeric protein in order to obtain a better understanding of the molecular basis of structure/function relationships. PMID- 19292864 TI - Role of calcium phosphate nanoclusters in the control of calcification. AB - Calcium phosphate nanoclusters are equilibrium particles of defined chemical composition in which a core of amorphous calcium phosphate is sequestered within a shell of casein phosphopeptides. Sequence analyses and a structure prediction method were applied to secreted phosphoproteins of known importance in controlling calcification, and eight noncasein phosphoproteins were identified as containing one or more subsequences capable of forming nanoclusters. Small-angle X-ray scattering was used to confirm that a plasmin phosphopeptide of one of the identified proteins, osteopontin, formed a novel type of calcium phosphate nanocluster in which the radius of the amorphous calcium phosphate core was four times larger than is typical of casein nanoclusters. A thermodynamic treatment of nanocluster formation identified the factors of importance in determining the equilibrium size of the core, and showed how a nanocluster solution could be thermodynamically stable yet supersaturated with respect to the mineral phase of bones and teeth. It is suggested that the ability of some secreted phosphoproteins to form nanoclusters is physiologically important for the control or inhibition of calcification in soft and mineralized tissues, the extracellular matrix and a wide range of biofluids, including milk and blood. PMID- 19292865 TI - Kinetics of inhibition of acetylcholinesterase in the presence of acetonitrile. AB - The hydrolysis of acetylthiocholine by acetylcholinesterase from Electrophorus electricus was investigated in the presence of the inhibitors tacrine, gallamine and compound 1. The interaction of the enzyme with the substrate and the inhibitors was characterized by the parameters K(I), alpha', b or beta, K(m) and V(max), which were determined directly and simultaneously from nonlinear Michaelis-Menten plots. Tacrine was shown to act as a mixed-type inhibitor with a strong noncompetitive component (alpha' approximately 1) and to completely block deacylation of the acyl-enzyme. In contrast, acetylcholinesterase inhibition by gallamine followed the 'steric blockade hypothesis', i.e. only substrate association to as well as substrate/product dissociation from the active site were reduced in the presence of the inhibitor. The relative efficiency of the acetylcholinesterase-gallamine complex for the catalysis of substrate conversion was determined to be 1.7-25% of that of the free enzyme. Substrate hydrolysis and the inhibition of acetylcholinesterase were also investigated in the presence of 6% acetonitrile, and a competitive pseudo-inhibition was observed for acetonitrile (K(I) = 0.25 m). The interaction of acetylcholinesterase with acetonitrile and tacrine or gallamine resulted in a seven- to 10-fold increase in the K(I) values, whereas the principal mode of inhibition was not affected by the organic solvent. The determination of the inhibitory parameters of compound 1 in the presence of acetonitrile revealed that the substance acts as a hyperbolic mixed-type inhibitor of acetylcholinesterase. The complex formed by the enzyme and the inhibitor still catalysed product formation with 8.7-9.6% relative efficiency. PMID- 19292866 TI - Cell biology, regulation and inhibition of beta-secretase (BACE-1). AB - Since the discovery of the beta-secretase responsible for initiating the Alzheimer's amyloid cascade as a novel membrane-bound aspartic proteinase, termed 'beta-site amyloid precursor protein cleaving enzyme', 'aspartyl protease-2' or 'membrane-anchored aspartic proteinase of the pepsin family-2', huge efforts have been devoted to an understanding of its biology and structure in the subsequent decade. This has paid off in many respects, as it has been cloned, its structure solved, novel physiological substrates of the enzyme discovered, and numerous inhibitors of its activity developed in a relatively short space of time. The inhibition of beta-secretase activity in vivo remains one of the most viable strategies for the treatment of Alzheimer's disease, although progress in getting inhibitors to the clinic has been slow, partly as a consequence of its aspartic proteinase character, which poses considerable problems for the production of potent, selective and brain-accessible compounds. This review reflects on the development of beta-secretase biology and chemistry to date, highlighting the diverse and innovative strategies applied to the modulation of its activity at the molecular and cellular levels. PMID- 19292867 TI - The PA-TM-RING protein RING finger protein 13 is an endosomal integral membrane E3 ubiquitin ligase whose RING finger domain is released to the cytoplasm by proteolysis. AB - PA-TM-RING proteins have an N-terminal protease-associated domain, a structure found in numerous proteases and implicated in protein binding, and C-terminal RING finger and PEST domains. Homologous proteins include GRAIL (gene related to anergy in leukocytes), which controls T-cell anergy, and AtRMR1 (receptor homology region-transmembrane domain-RING-H2 motif protein), a plant protein storage vacuole sorting receptor. Another family member, chicken RING zinc finger (C-RZF), was identified as being upregulated in embryonic chicken brain cells grown in the presence of tenascin-C. Despite algorithm predictions that the cDNA encodes a signal peptide and transmembrane domain, the protein was found in the nucleus. We showed that RING finger protein 13 (RNF13), the murine homolog of C RZF, is a type I integral membrane protein localized in the endosomal/lysosomal system. By quantitative real-time RT-PCR analysis, we demonstrated that expression of RNF13 is increased in adult relative to embryonic mouse tissues and is upregulated in B35 neuroblastoma cells stimulated to undergo neurite outgrowth. We found that RNF13 is very labile, being subject to extensive proteolysis that releases both the protein-associated domain and the RING domain from the membrane. By analyzing microsomes, we showed that the ectodomain is shed into the lumen of vesicles, whereas the C-terminal half, which possesses the RING finger, is released to the cytoplasm. This C-terminal fragment of RNF13 has the ability to mediate ubiquitination. Proteolytic release of RNF13 from a membrane anchor thus provides unique spatial and temporal regulation that has not been previously described for an endosomal E3 ubiquitin ligase. PMID- 19292868 TI - Novel genes in cell cycle control and lipid metabolism with dynamically regulated binding sites for sterol regulatory element-binding protein 1 and RNA polymerase II in HepG2 cells detected by chromatin immunoprecipitation with microarray detection. AB - Sterol regulatory element-binding proteins 1 and 2 (SREBP-1 and SREBP-2) are important regulators of genes involved in cholesterol and fatty acid metabolism, but have also been implicated in the regulation of the cell cycle and have been associated with the pathogenesis of type 2 diabetes, atherosclerosis and obesity, among others. In this study, we aimed to characterize the binding sites of SREBP 1 and RNA polymerase II through chromatin immunoprecipitation and microarray analysis in 1% of the human genome, as defined by the Encyclopaedia of DNA Elements consortium, in a hepatocellular carcinoma cell line (HepG2). Our data identified novel binding sites for SREBP-1 in genes directly or indirectly involved in cholesterol metabolism, e.g. apolipoprotein C-III (APOC3). The most interesting biological findings were the binding sites for SREBP-1 in genes for host cell factor C1 (HCFC1), involved in cell cycle regulation, and for filamin A (FLNA). For RNA polymerase II, we found binding sites at classical promoters, but also in intergenic and intragenic regions. Furthermore, we found evidence of sterol-regulated binding of SREBP-1 and RNA polymerase II to HCFC1 and FLNA. From the results of this work, we infer that SREBP-1 may be involved in processes other than lipid metabolism. PMID- 19292869 TI - Proteolytic processing of porcine deoxyribonuclease II occurs in lysosomes but is not required for enzyme activation. AB - DNase II purified from porcine spleen (pDNase II) comprises alpha(1), alpha(2) and beta subunits. The three subunits are encoded by one cDNA, in the sequence alpha(1), beta, and alpha(2), and the peptides linking these subunits are presumably cleaved out post-translationally. To understand the relevance of post translational cleavage to pDNase II, recombinant pDNase II (rpDNase II) was produced in human 293T cells by transfection with an expression plasmid containing pDNase II cDNA (pcDNaseII). An 11.5, a 35 and a 46.5 kDa protein were detected in the cell lysates, whereas only a 46.5 kDa protein was detected in the culture medium of the pcDNaseII-transfected cells. The 46.5 kDa rpDNase II secreted into the medium was purified to homogeneity and characterized. MALDI-TOF MS and N-terminal amino acid sequencing of the 46.5 kDa protein revealed a single contiguous polypeptide chain of pDNase II. Zymographic analysis showed that the 46.5 kDa protein digested DNA in acidic conditions and that the specific activity of this rpDNase II was about twice that of pDNase II purified from porcine spleen. Treatment with chloroquine, a lysosomal inhibitor, resulted in the accumulation of only the 46.5 kDa protein in the pcDNaseII-transfected cells. Treatments with cycloheximide 22 h after transfection led to accumulation of the processed enzyme and disappearance of the 46.5 kDa protein. These results suggest that the proteolytic processing of rpDNase II occurs in the lysosome, which is not involved in the activation of pDNase II. PMID- 19292870 TI - Alpha-1 antitrypsin binds preprohepcidin intracellularly and prohepcidin in the serum. AB - Recent discoveries have indicated that the hormone hepcidin plays a major role in the control of iron homeostasis. Hepcidin regulates the iron level in the blood through the interaction with ferroportin, an iron exporter molecule, causing its internalization and degradation. As a result, hepcidin increases cellular iron sequestration, and decreases the iron concentration in the plasma. Only mature hepcidin (result of the cleavage of prohepcidin by furin proteases) has biological activity; however, prohepcidin, the prohormone form, is also present in the plasma. In this study, we aimed to identify new protein-protein interactions of preprohepcidin, prohepcidin and hepcidin using the BacterioMatch two-hybrid system. Screening assays were carried out on a human liver cDNA library. Preprohepcidin screening gave the following results: alpha-1 antitrypsin, transthyretin and alpha-1-acid glycoprotein showed strong interactions with preprohepcidin. We further confirmed and examined the alpha-1 antitrypsin binding in vitro (glutathione S-transferase, pull down, coimmunoprecipitation, MALDI-TOF) and in vivo (ELISA, cross-linking assay). Our results demonstrated that the serine protease inhibitor alpha-1 antitrypsin binds preprohepcidin within the cell during maturation. Furthermore, alpha-1 antitrypsin binds prohepcidin significantly in the plasma. This observation may explain the presence of prohormone in the circulation, as well as the post translational regulation of the mature hormone level in the blood. In addition, the lack of cleavage protection in patients with alpha-1 antitrypsin deficiency may be the reason for the disturbance in their iron homeostasis. PMID- 19292871 TI - Chronic high-dose morphine treatment promotes SH-SY5Y cell apoptosis via c-Jun N terminal kinase-mediated activation of mitochondria-dependent pathway. AB - Chronic high doses of morphine inhibit the growth of various human cancer cell lines. However, the mechanisms by which such high-dose morphine inhibits cell proliferation and induces cell death are not fully understood. Here we show that c-Jun N-terminal kinase (JNK) plays a pivotal role in high-dose morphine-induced apoptosis of SH-SY5Y cells in a mitochondria-dependent manner. Activation of JNK by morphine led to reactive oxygen species (ROS) generation via the mitochondrial permeability transition pore, because the mPTP inhibitor cyclosporin A significantly inhibited ROS generation. ROS in turn exerted feedback regulation on JNK activation, as shown by the observations that cyclosporin A and the antioxidant N-acetylcysteine significantly inhibited the phosphorylation of JNK induced by morphine. ROS-amplified JNK induced cytochrome c release and caspase 9/3 activation through enhancement of expression of the proapoptotic protein Bim and reduction of expression of the antiapoptotic protein Bcl-2. All of these effects of morphine could be suppressed by the JNK inhibitor SP600125 and N acetylcysteine. The key role of the JNK pathway in morphine-induced apoptosis was further confirmed by the observation that decreased levels of JNK in cells transfected with specific small interfering RNA resulted in resistance to the proapoptotic effect of morphine. Thus, the present study clearly shows that morphine-induced apoptosis in SH-SY5Y cells involves JNK-dependent activation of the mitochondrial death pathway, and that ROS signaling exerts positive feedback regulation of JNK activity. PMID- 19292872 TI - Molecular basis of the unusual catalytic preference for GDP/GTP in Entamoeba histolytica 3-phosphoglycerate kinase. AB - Phosphoglycerate kinase (EC 2.7.2.3) catalyzes reversible phosphoryl transfer from 1,3-bisphosphoglycerate to ADP to synthesize 3-phosphoglycerate and ATP during glycolysis. Phosphoglycerate kinases from several sources can use GDP/GTP as alternative substrates to ADP/ATP; however, the maximal velocities (V(m)) reached with the guanine nucleotides are approximately 50% of those displayed with the adenine nucleotides. By contrast, Entamoeba histolytica phosphoglycerate kinase (EC 2.7.2.10) is the only reported phosphoglycerate kinase displaying higher activity with GDP/GTP and lower affinities for the adenine nucleotides. To elucidate the molecular basis of the Entamoeba histolytica phosphoglycerate kinase selectivity for GDP/GTP, a conformational analysis was carried out on a homology model based on crystallographic structures of yeast and pig phosphoglycerate kinases. Some amino acid residues involved in the purine ring binding site not previously described were detected. Accordingly, Y239, E309 and V311 were replaced by site-directed mutagenesis in the Entamoeba histolytica phosphoglycerate kinase gene for the corresponding amino acid residues present in the adenine nucleotide-dependent phosphoglycerate kinases and the recombinant proteins were purified. Kinetic analysis of the enzymes showed that the single mutants Y239F, E309Q, E309M and V311L increased their catalytic efficiencies (V(m)/K(m)) with ADP/ATP as a result of both, increased V(m) and decreased K(m) values. Furthermore, a higher catalytic efficiency in the double mutant Y239F/E309M was achieved, which was mainly due to an increased affinity for ADP/ATP with a concomitant diminished affinity for GDP/GTP. The main Entamoeba histolytica phosphoglycerate kinase amino acid residues involved in the selectivity for guanine nucleotides were thus identified. PMID- 19292873 TI - Functional and structural characterization of novel mutations and genotype phenotype correlation in 51 phenylalanine hydroxylase deficient families from Southern Italy. AB - Hyperphenylalaninemia (Online Mendelian Inheritance in Man database: 261600) is an autosomal recessive disorder mainly due to mutations in the gene for phenylalanine hydroxylase; the most severe form of hyperphenylalaninemia is classic phenylketonuria. We sequenced the entire gene for phenylalanine hydroxylase in 51 unrelated hyperphenylalaninemia patients from Southern Italy. The entire locus was genotyped in 46 out of 51 hyperphenylalaninemia patients, and 32 different disease-causing mutations were identified. The pathologic nature of two novel gene variants, namely, c.707-2delA and p.Q301P, was demonstrated by in vitro studies. c.707-2delA is a splicing mutation that involves the accepting site of exon 7; it causes the complete skipping of exon 7 and results in the truncated p.T236MfsX60 protein. The second gene variant, p.Q301P, has very low residual enzymatic activity (approximately 4.4%), which may be ascribed, in part, to a low expression level (8-10%). Both the decreased enzyme activity and the low expression level are supported by analysis of the 3D structure of the molecule. The putative structural alterations induced by p.Q301P are compatible with protein instability and perturbance of monomer interactions within dimers and tetramers, although they do not affect the catalytic site. In vivo studies showed tetrahydrobiopterin responsiveness in the p.Q301P carrier but not in the c.707 2delA carrier. We next investigated genotype-phenotype correlations and found that genotype was a good predictor of phenotype in 76% of patients. However, genotype-phenotype discordance occurred in approximately 25% of our patients, mainly those bearing mutations p.L48S, p.R158Q, p.R261Q and p.P281L. PMID- 19292874 TI - Low U1 snRNP dependence at the NF1 exon 29 donor splice site. AB - Many disease-causing splicing mutations described in the literature produce changes in splice sites (SS) or in exon-regulatory sequences. The delineation of these splice aberrations can provide important insights into novel regulation mechanisms. In this study, we evaluated the effect of patient variations in neurofibromatosis type 1 (NF1) exon 29 and its 5'SS surrounding area on its splicing process. Only two of all nonsense, missense, synonymous and intronic variations analyzed in this study clearly altered exon 29 inclusion/exclusion levels. In particular, the intronic mutation +5g>a had the strongest effect, resulting in total exon exclusion. This finding prompted us to evaluate the exon 29 5'SS in relation to its ability to bind U1 snRNP. This was performed by direct analysis of the ability of U1 to bind to wild-type and mutant donor sites, by engineering an in vitro splicing system to directly evaluate the functional importance of U1 snRNA base pairing with the exon 29 donor site, and by coexpression of mutant U1 snRNP molecules to try to rescue exon 29 inclusion in vivo. The results revealed a low dependency on the presence of U1 snRNP, and suggest that exon 29 donor site definition may depend on alternative mechanisms of 5'SS recognition. PMID- 19292875 TI - Hydrolysis of acetylthiocoline, o-nitroacetanilide and o nitrotrifluoroacetanilide by fetal bovine serum acetylcholinesterase. AB - Besides esterase activity, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) hydrolyze o-nitroacetanilides through aryl acylamidase activity. We have reported that BuChE tetramers and monomers of human blood plasma differ in o nitroacetanilide (ONA) hydrolysis. The homology in quaternary structure and folding of subunits in the prevalent BuChE species (G4(H)) of human plasma and AChE forms of fetal bovine serum prompted us to study the esterase and amidase activities of fetal bovine serum AChE. The k(cat)/K(m) values for acetylthiocholine (ATCh), ONA and its trifluoro derivative N-(2-nitrophenyl) trifluoroacetamide (F-ONA) were 398 x 10(6) M(-1) min(-1), 0.8 x 10(6) M(-1) min( 1), and 17.5 x 10(6) M(-1) min(-1), respectively. The lack of inhibition of amidase activity at high F-ONA concentrations makes it unlikely that there is a role for the peripheral anionic site (PAS) in F-ONA degradation, but the inhibition of ATCh, ONA and F-ONA hydrolysis by the PAS ligand fasciculin-2 points to the transit of o-nitroacetalinides near the PAS on their way to the active site. Sedimentation analysis confirmed substrate hydrolysis by tetrameric 10.9S AChE. As compared with esterase activity, amidase activity was less sensitive to guanidine hydrochloride. This reagent led to the formation of 9.3S tetramers with partially unfolded subunits. Their capacity to hydrolyze ATCh and F-ONA revealed that, despite the conformational change, the active site architecture and functionality of AChE were partially retained. PMID- 19292876 TI - TICL--a web tool for network-based interpretation of compound lists inferred by high-throughput metabolomics. AB - High-throughput metabolomics is a dynamically developing technology that enables the mass separation of complex mixtures at very high resolution. Metabolic profiling has begun to be widely used in clinical research to study the molecular mechanisms of complex cell disorders. Similar to transcriptomics, which is capable of detecting genes at differential states, metabolomics is able to deliver a list of compounds differentially present between explored cell physiological conditions. The bioinformatics challenge lies in a statistically valid interpretation of the functional context for identified sets of metabolites. Here, we present TICL, a web tool for the automatic interpretation of lists of compounds. The major advance of TICL is that it not only provides a model of possible compound transformations related to the input list, but also implements a robust statistical framework to estimate the significance of the inferred model. The TICL web tool is freely accessible at http://mips.helmholtz muenchen.de/proj/cmp. PMID- 19292877 TI - NMR studies on the interaction of sugars with the C-terminal domain of an R-type lectin from the earthworm Lumbricus terrestris. AB - The R-type lectin EW29, isolated from the earthworm Lumbricus terrestris, consists of two homologous domains (14,500 Da) showing 27% identity with each other. The C-terminal domain (Ch; C-half) of EW29 (EW29Ch) has two sugar-binding sites in subdomains alpha and gamma, and the protein uses these sugar-binding sites for its function as a single-domain-type hemagglutinin. In order to determine the sugar-binding ability and specificity for each of the two sugar binding sites in EW29Ch, ligand-induced chemical-shift changes in EW29Ch were monitored using (1)H-(15)N HSQC spectra as a function of increasing concentrations of lactose, melibiose, D-galactose, methyl alpha-D galactopyranoside and methyl beta-D-galactopyranoside. Shift perturbation patterns for well-resolved resonances confirmed that all of these sugars associated independently with the two sugar-binding sites of EW29Ch. NMR titration experiments showed that the sugar-binding site in subdomain alpha had a slow or intermediate exchange regime on the chemical-shift timescale (K(d) = 10( 2) to 10(-1) mM), whereas that in subdomain gamma had a fast exchange regime for these sugars (K(d) = 2-6 mM). Thus, our results suggest that the two sugar binding sites of EW29Ch in the same molecule retain its hemagglutinating activity, but this activity is 10-fold lower than that of the whole protein because EW29Ch has two sugar-binding sites in the same molecule, one of which has a weak binding mode. PMID- 19292878 TI - Acute intermittent porphyria--impact of mutations found in the hydroxymethylbilane synthase gene on biochemical and enzymatic protein properties. AB - Acute intermittent porphyria is an autosomal dominantly inherited disorder, classified as acute hepatic porphyria, caused by a deficiency of hydroxymethylbilane synthase (EC 2.5.1.61, EC 4.3.1.8, also known as porphobilinogen deaminase, uroporphyrinogen I synthase), the third enzyme in heme biosynthesis. Clinical features include autonomous, central, motor or sensory symptoms, but the most common clinical presentation is abdominal pain caused by neurovisceral crises. A diagnosis of acute intermittent porphyria is crucial to prevent life-threatening acute attacks. Detection of DNA variations by molecular techniques allows a diagnosis of acute intermittent porphyria in situations where the measurement of porphyrins and precursors in urine and faeces and erythrocyte hydroxymethylbilane synthase activity is inconclusive. In the present study, we identified gene defects in six Czech patients with acute intermittent porphyria, as diagnosed based on biochemical findings, and members of their families to confirm the diagnosis at the molecular level and/or to provide genetic counselling. Molecular analyses of the hydroxymethylbilane synthase gene revealed seven mutations. Four were previously reported: c.76C>T, c.77G>A, c.518G>A, c.771 + 1G>T (p.Arg26Cys, p.Arg26His, p.Arg173Gln). Three were novel mutations: c.610C>A, c.675delA, c.750A>T (p.Gln204Lys, p.Ala226ProfsX28, p.Glu250Asp). Of particular interest, one patient had two mutations (c.518G>A; c.610C>A), both located in exon 10 of the same allele. To establish the effects of the mutations on enzyme function, biochemical characterization of the expressed normal recombinant and mutated proteins was performed. Prokaryotic expression of the mutant alleles of the hydroxymethylbilane synthase gene revealed that, with the exception of the p.Gln204Lys mutation, all mutations resulted in little, if any, enzymatic activity. Moreover, the 3D structure of the Escherichia coli and human protein was used to interpret structure-function relationships for the mutations in the human isoform. PMID- 19292879 TI - X-ray structure of glucose/galactose receptor from Salmonella typhimurium in complex with the physiological ligand, (2R)-glyceryl-beta-D-galactopyranoside. AB - Periplasmic binding proteins are abundant in bacteria by virtue of their essential roles as high-affinity receptors in ABC transport systems and chemotaxis. One of the best studied of these receptors is the so-called glucose/galactose-binding protein. Here, we report the X-ray structure of the Salmonella typhimurium protein bound to the physiologically relevant ligand, (2R) glyceryl-beta-D-galactopyranoside, solved by molecular replacement, and refined to 1.87 A resolution with R and R-free values of 17% and 22%. The structure identifies three amino acid residues that are diagnostic of (2R)-glyceryl-beta-D galactopyranoside binding (Thr110, Asp154 and Gln261), as opposed to binding to the monosaccharides glucose and galactose. These three residues are conserved in essentially all available glucose/galactose-binding protein sequences, indicating that the binding of (2R)-glyceryl-beta-D-galactopyranoside is the rule rather than the exception for receptors of this type. The role of (2R)-glyceryl-beta-D galactopyranoside in bacterial biology is discussed. Further, comparison of the available structures provides the most complete description of the conformational changes of glucose/galactose-binding protein to date. The structures follow a smooth and continuous path from the most closed structure [that bound to (2R) glyceryl-beta-D-galactopyranoside] to the most open (an apo structure). PMID- 19292880 TI - Mechanism of mild acid hydrolysis of galactan polysaccharides with highly ordered disaccharide repeats leading to a complete series of exclusively odd-numbered oligosaccharides. AB - Sulfated galactan kappa-carrageenan is a linear polysaccharide with a repeating disaccharide sequence of alternating 4-sulfated 3-linked galactose and 4-linked 3,6-anhydrogalactose units. In contrast to many examples of chemical hydrolysis of polysaccharides, mild acid treatment of kappa-carrageenan resulted in facile and highly specific cleavage. In this article, we report the identification, by various MS and chromatographic techniques, of an unexpected series of exclusively odd-numbered oligosaccharide fragments from its hydrolytic products. Detailed sequence analysis of the products indicated that all the oligosaccharide fragments have the 4-sulfated 3-linked galactose residues at both the reducing and the nonreducing ends. Further detailed investigation and analysis suggested that these odd-numbered oligosaccharides were derived from two-step cleavages of the glycosidic bonds on either sides of the 3,6-anhydrogalactose residues. Neutral galactan agarose also contains 3,6-anhydrogalactose and has a similar backbone sequence, and exhibited similar results upon mild acid hydrolysis. It is highly unusual to obtain exclusively odd-numbered oligosaccharides from polysaccharides composed of ordered disaccharide repeats. PMID- 19292881 TI - Glutathione, vitamin E and oxidative stress in coronary artery disease: relevance of age and gender. AB - BACKGROUND: Observational studies suggest that low levels of antioxidants are associated with high risk for coronary artery disease (CAD). We investigated whether the biomarkers of oxidative balance undergo the same modifications in all CAD patient groups, regardless of gender and age. MATERIALS AND METHODS: One hundred sixty-eight CAD patients and 107 healthy controls were assayed for plasma levels of reduced glutathione (GSH), alpha- and gamma-tocopherol (alpha- and gamma-T) as endogenous antioxidants. A damage score (DS), representative of oxidative stress status, was calculated. ANCOVA models were used to test the association between antioxidants, DS and CAD and its modulation by age and gender. RESULTS: The DS was higher in CAD than in controls. GSH levels, were lower in CAD patients (mean +/- SEM: 57.61 +/- 1.87 micromol 10 g(-1) haemoglobin vs. 68.55 +/- 2.23 in controls, P < 0.0006) in males and in older subjects. Levels of other antioxidants exhibited a complex pattern. Overall, no difference was found in alpha- and gamma-T contents between CAD and controls, but lower alpha-T values were observed in CAD females. A significant interaction between CAD status and gender was observed (P = 0.003). CONCLUSIONS: Our study shows that the involvement of antioxidants in CAD is related to patients' characteristics. These findings may be relevant in planning antioxidant therapies. PMID- 19292882 TI - Predictors of inotropic and chronotropic effects of NG-monomethyl-L-arginine. AB - BACKGROUND: The haemodynamic effects of intravenous infusion of the non-selective nitric oxide synthase (NOS) L-omega monomethyl arginine (L-NMMA) have previously been characterized in humans. Its effect of reducing cardiac index (CI) is an important reason for the increase in mortality in patients with septic shock receiving L-NMMA in a pivotal outcome trial for this indication. The mechanism for the reduction in CI however, is uncertain. METHODS: In this study, we investigated the haemodynamic and arterial stiffness response to a bolus intravenous infusion of L-NMMA (3 mg kg(-1) over 5 min) in 26 healthy human volunteers to clarify the likely cause of L-NMMA induced negative inotropic and chronotropic effects. Digital photoplethysmography (MicroMedical Pulse Trace) was used to derive two measures of arterial stiffness: stiffness index, a measure of large arterial stiffness, and reflection index (RI), a measure of small- to medium-sized arterial stiffness. Haemodynamic measurements of systolic blood pressure, diastolic blood pressure, heart rate, systemic vascular resistance index (SVRI), stroke index and CI were made using a bioimpedance monitor (BioZ Cardiodynamics). RESULTS: We found that changes in CI during L-NMMA are closely related to changes in RI and SVRI. CONCLUSION: The negative inotropic effect of L NMMA may be a result of an increase in coronary vascular resistance and a resultant decrease in myocardial perfusion. The reduction in CI may also result from a direct reduction of the normal positive inotropic effect of NO by L-NMMA which is closely correlated with its effects on SVRI. PMID- 19292883 TI - TNF-alpha modulates the Na+/ K+ ATPase and the Na+K+2Cl- symporter in LLC-PK cells. AB - BACKGROUND: Tumour necrosis factor alpha (TNF-alpha) has been implicated in the development of diabetic nephropathy and the accompanying increase in sodium retention. Inhibition of renal Na(+)/K(+) ATPase was reported to accompany cell death. As TNF is known to induce both apoptosis and cell survival, this work investigated the effect and mechanism of action of TNF-alpha on the Na(+)/K(+) ATPase and the Na(+)K(+)2Cl(-) symporter using LLC-PK(1) cells, a porcine renal proximal tubules cell line. MATERIALS AND METHODS: Cells were incubated for 2 h with TNF-alpha in presence and absence of pyrrolidinedithiocarbamate, SP600125 and FK009, respective inhibitors of the transcription factor nuclear factor kappa light-chain-enhancer of activated B cells (NF-kappaB), c-Jun N-terminal kinase (JNK) and caspases. The activity of the pump was assayed by measuring the ouabain inhibitable release of inorganic phosphate. Changes in its expression and the expression of the symporter were monitored by western blot analysis. RESULTS: TNF alpha up-regulated both transporters. NF-kappaB, JNK and the caspases were all mediators of the cytokine action. TNF up-regulated the Na(+)/K(+) pump by stimulating JNK which in turn, activated NF-kappaB and inhibited the caspases. TNF effect on the cotransporter was also mediated via activation of JNK which however inhibited NF-kappaB and by so doing prevented activation of caspases. As caspases were demonstrated to down-regulate the two transporters, their inhibition by TNF is responsible for the observed up-regulatory effect. CONCLUSIONS: It was concluded that the Na(+)/K(+) ATPase and Na(+)K(+)2Cl(-) are both targets of TNF-alpha and the effect of the cytokine favours cell survival over cell death. PMID- 19292884 TI - Chronic renal failure induces genetic instability in multiple organs of Wistar rats. AB - BACKGROUND: Taking into consideration the strong evidence for a relationship between DNA damage and carcinogenesis, the aim of this study was to investigate whether blood, liver, heart, kidney and brain are particularly sensitive organs for DNA damaging during chronic renal disease by the single-cell gel (comet) assay to predict genetic instability induced by this pathological condition. METHODS: A total of 18 male Wistar rats were divided into two groups: negative control (n = 8) and experimental (n = 10), in which was submitted to the 5/6 renal mass ablation by ligation of two or three branches of the left renal artery and total right nephrectomy during 8 weeks. RESULTS: The results showed that chronic renal disease was able to induce genetic damage in blood, heart, liver and kidney cells as depicted by the mean tail moment. No genetic damage was induced in brain cells, i.e. no significant statistically differences (P > 0.05) were noticed when compared to negative control. CONCLUSION: In conclusion, our results suggest that chronic renal failure could contribute to the damage of DNA at all organs evaluated, except to the brain cells. As DNA damage is an important step in events leading to carcinogenesis, this study represents a relevant contribution to the correct evaluation of the potential health risks associated with kidney disease. PMID- 19292885 TI - Simvastatin increases osteoblasts and osteogenic proteins in ovariectomized rats. AB - BACKGROUND: Previous reports have indicated that statins could prevent bone loss in ovariectomized (OVX) rats and increase the expressions of osteogenic genes in cultured osteoblasts. In this study, we hypothesized that simvastatin might increase osteoblast number and protein expressions of osteogenic markers localized in bones in concomitance with the prevention of bone loss in OVX rats. MATERIALS AND METHODS: Fifty-four 3-month-old OVX and sham-operated (SHAM) female Sprague-Dawley rats were used. Simvastatin (10-20 mg kg(-1) day(-1)) was administrated orally for 6 weeks. Trabecular volume, osteoblast number and osteogenic proteins including BMP2, collagen type I and osteocalcin on bone sections obtained from lumbar vertebral body, distal femur and proximal tibia were measured. RESULTS: The results showed that SHAM rats had significantly less trabecular bone volume and osteoblast number than that of OVX rats 6 weeks after operation. Oral simvastatin treatment (10-20 mg kg(-1) day(-1)) increased bone volume and osteoblast number in the distal femurs, proximal tibiae and vertebrae of OVX rats. Furthermore, the osteoblastic cells with immuno-stained BMP2, collagen type I and osteocalcin in vertebral bones were significantly increased by simvastatin treatment (20 mg kg(-1) day(-1)) in OVX rats. CONCLUSIONS: This study demonstrates that simvastatin enhances the production of osteogenic proteins in bone and this effect may contribute to the prevention of bone loss in OVX rats. PMID- 19292886 TI - CD98hc (SLC3A2), a novel marker in renal cell cancer. AB - BACKGROUND: In a variety of malignant diseases, molecular targeting represents a therapeutic option, whereby, when compared with chemotherapy, fewer side effects are thought to be expected. Especially in renal cell cancer (RCC), tyrosine kinase-inhibitors have been established as useful and highly effective therapy. However, tyrosine kinase-inhibitors currently approved for RCC treatment lack single molecule specificity and bear a variety of side effects of the gastro intestinal tract, skin, heart and haematopoietic system. Therefore, the identification of novel cell surface markers is sought, which might lead to novel diagnostic and therapeutic strategies in cancer. MATERIAL AND METHODS: Paraffin embedded RCCs from a well characterized tissue bank were immunohistochemically quantified for embryonic transmembrane antigen CD98hc (SLC3A2) expression and semi-quantitative analyses were correlated with subtype or grade of differentiation. RESULTS: We found increased CD98hc expression in different types of malign RCCs, among them clear cell (cc)RCC, papillary (p)RCC and chromophobe (ch)RCC, but lack of expression in the benign renal oncocytoma. Thereby, the extent of CD98hc expression directly complies with grade of malignancy. Furthermore, the more malignant type II pRCC significantly higher expressed CD98hc than the less malignant and more differentiated type I pRCC (type II 83.34%, type I 4.76% CD98hc positive, P < 0.00001; n = 51). The established marker for type I pRCC, Cytokreatin 7, showed 95.24% expression in type I and 26.67% expression in type II pRCC (P < 0.00001, n = 51). CONCLUSIONS: From these data, we conclude that CD98hc is expressed in RCCs, whereby the extent of expression is likely to correlate directly with grade of malignancy. In pRCCs, CD98hc might represent a novel and reliable marker for type II pRCC. PMID- 19292887 TI - Six different CYBA mutations including three novel mutations in ten families from Turkey, resulting in autosomal recessive chronic granulomatous disease. AB - BACKGROUND: One of the rarest forms of autosomal recessive chronic granulomatous disease (AR-CGD) is attributable to mutations in the CYBA gene, which encodes the alpha polypeptide of cytochrome b(558), (also known as p22-phox), a key transmembrane protein in the phagocyte NADPH oxidase system. This gene is localized on chromosome 16q24, encompasses 8.5 kb and contains six exons. MATERIALS AND METHODS: We report here the clinical and molecular characterization of 12 AR-CGD patients from 10 consanguineous, unrelated Turkish families with clinical CGD and positive family history. The ages of the six male and six female patients were between 1and 18 years. Before mutation analysis, subgroup analysis of patients was made by flow cytometry with antibodies against NADPH-oxidase components and with the DHR assay (flow cytometric assay of NADPH oxidase activity in leucocytes). RESULTS: Mutation analysis of CYBA showed six different mutations: a frameshift insertion in exon 3 (C after C166); a missense mutation in exon 2 (p.Gly24Arg), a splice-site deletion in intron 1 (4-bp deletion +4_+7 AGTG), a novel nonsense mutation in exon 6 (p.Cys113X), a novel large deletion of exons 3-6 and a novel 1-bp deletion in exon 6 (c.408delC). All mutations were present in homozygous form and all parents investigated were found to be heterozygotes for these mutations. CONCLUSIONS: In our series of 40 CGD families, approximately 25% of the families have p22-phox defects, with six different mutations, including three novel mutations. The high rate of consanguineous marriages seems to be the underlying aetiology. PMID- 19292888 TI - PDGF-C and -D and their receptors PDGFR-alpha and PDGFR-beta in atherosclerotic human arteries. AB - BACKGROUND: Platelet derived growth factors (PDGFs) are mitogens for fibroblasts and smooth muscle cells. This growth factor family contains four members PDGF-A, PDGF-B, PDGF-C and PDGF-D. Biology of recently discovered PDGF-C and PDGF-D is not well-established. Here we studied the expression of PDGF-C and PDGF-D and their receptors PDGFR-alpha and PDGFR-beta in normal and atherosclerotic human arteries. MATERIALS AND METHODS: Human arterial samples from amputations and autopsies were classified according to the atherosclerotic stage and the expression of PDGF-C and PDGF-D proteins and their receptors was studied by immunohistochemistry. In situ hybridization and reverse transcriptase-PCR were used to study mRNA expression. RESULTS: Both growth factors were expressed in medial smooth muscle cells (SMCs) in normal arteries and atherosclerotic lesions. However, clear differences were found in the expression profiles in endothelium: PDGF-C was strongly expressed in endothelial cells in both normal arteries and lesions whereas PDGF-D was only weakly expressed in endothelium. PDGF-C expression was very prominent in lesion macrophages. PDGF-D was expressed throughout the artery wall in lesions. PDGFR-alpha expression was strong in endothelium and in lesion macrophage-rich areas, whereas PDGFR-beta was mostly expressed in SMCs. CONCLUSIONS: Our results suggest that PDGF-C may play an important role in endothelium in normal and atherosclerotic arteries and in macrophages in lesions. PDGF-D was expressed in all types of lesions with the same intensity and thus differs from the expression of PDGF-C. PMID- 19292889 TI - Lysyl oxidase like-1 dysregulation and its contribution to direct inguinal hernia. AB - OBJECTIVE: The aetiology of inguinal hernia involves changes in collagen turnover and metalloproteinase expression; yet it is not known whether the elastic fibre system could also be affected. This study was designed to compare the expression of tropoelastin (TE), lysyl oxidase-like 1 (LOXL-1) and elastase in the transversalis fascia of patients with and without inguinal hernia. MATERIAL AND METHODS: Transversalis fascia (TF) specimens were obtained from patients undergoing surgery for direct or indirect inguinal hernia (n = 20 each) and from multi-organ donors during organ procurement (controls, n = 16). The specimens were divided according to age (20-40/41-60 years). Tissues were immunohistochemically labelled using anti-tropoelastin, anti-LOXL-1 and anti elastase antibodies and subjected to Western blot analysis. Relative amounts of LOXL-1 and TE mRNA were determined by real time RT-PCR in cultured cells obtained from the TF of patients and controls. RESULTS: Significantly lower TE and LOXL-1 levels were observed in patients with direct inguinal hernia compared with controls or those with indirect hernia. In contrast, patients with direct inguinal hernia showed significantly higher elastase expression. In fibroblasts isolated from the TF, relative amounts of tropoelastin mRNA were lower for the hernia groups but differences were not significant. LOXL-1 mRNA levels were significantly lower in the direct hernia group compared to controls. CONCLUSIONS: Our findings suggest that impaired elastic fibre function in the transversalis fascia of patients with direct inguinal hernia, reflected by diminished elastin synthesis and its enhanced enzyme degradation, contributes to the development of this type of hernia. PMID- 19292890 TI - Downregulation of developmentally regulated endothelial cell locus-1 inhibits the growth of colon cancer. AB - Developmentally regulated endothelial cell locus-1 (Del1) is an embryonic angiogenic factor expressed in early embryonic endothelial cells, but recently has been found to be expressed in some forms of cancers including colon and breast cancers, and melanoma, and human cancer cell lines. Overexpression of Del1 accelerates tumor growth by enhancing vascular formation, implying Del1 may be a potential target for anti-angiogenic cancer therapy. The study aims to investigate whether downregulation of Del1 could inhibit the growth of tumors established in nude Balb/c mice by subcutaneous implantation of human LS-174T colon cancer cells. The shRNA expression vectors targeting human Del1, and vascular endothelial growth factor (VEGF) were constructed. Gene transfection of Del1-shRNA downregulated expression of Del1 in LS-174T cells in vivo and in vitro, but did not alter the proliferative or survival properties of cells in vitro. Gene transfection of VEGF-shRNA downregulated expression of both VEGF and Del1 in LS-174T cells in vivo and in vitro. Both Del1-shRNA and VEGF-shRNA gene therapies exhibited anti-tumor activities and they also showed a synergistic effect in suppressing growth of colon tumors by anti-angiogenesis and anti proliferation. Although further investigation to clarify the mechanisms explaining the role of Del1 in tumor growth, and the interaction between VEGF and Del1, is required, the results indicate that downregulation of Del1 presents a potent therapeutic strategy to combat colon cancer. PMID- 19292891 TI - Persistent H. pylori colonization in early acquisition age of mice related with higher gastric sialylated Lewis x, IL-10, but lower interferon-gamma expressions. AB - BACKGROUND: H. pylori infection is less prevalent in childhood. This study validated whether the rates of H. pylori colonization depend on different acquisition ages, and correlate with the different gastric Lewis antigens or cytokine expressions after H. pylori acquisition. METHODS: We applied a young (7 day-old) C57BL/6 mice group (n = 50) and adult (6-week-old) C57BL/6 mice group (n = 50). In each group, 30 mice were challenged with H. pylori and 20 mice served as naive control. The success of H. pylori colonization was assessed on the 2(nd )week and the 8(th )week, respectively. The intensity of the Lewis x, sialylated Lewis x(sialyl-Le(x)), and cytokine expressions, including TNF-alpha, IFN-gamma, IL-6, IL-10, and IL-1beta, were immunochemically stained and graded. RESULTS: On the 2(nd )week after H. pylori challenge, the colonization rates of H. pylori were similar between the young mice group and the adult mice group (89% vs. 100%, P > 0.05). However, on the 8(th )week, the H. pylori colonization rate was significantly lower in the young mice group than in the adult mice group (53% vs. 95%, P = 0.003). On the 8(th )week, the young mice with a persistence of H. pylori colonization had higher sialyl-Le(x), higher IL-10, and lower IFN-gamma than those of the mice that lost colonization during the 2(nd )to the 8(th )week (P < 0.05). CONCLUSION: The persistence of H. pylori colonization could be an acquisition-age determinant process. After H. pylori exposure at an early acquisition age, the host response with a higher sialyl-Le(x )and IL-10, but a lower IFN-gamma correlates to the consequent persistence of H. pylori colonization. PMID- 19292892 TI - Brains swinging in concert: cortical phase synchronization while playing guitar. AB - BACKGROUND: Brains interact with the world through actions that are implemented by sensory and motor processes. A substantial part of these interactions consists in synchronized goal-directed actions involving two or more individuals. Hyperscanning techniques for assessing fMRI simultaneously from two individuals have been developed. However, EEG recordings that permit the assessment of synchronized neuronal activities at much higher levels of temporal resolution have not yet been simultaneously assessed in multiple individuals and analyzed in the time-frequency domain. In this study, we simultaneously recorded EEG from the brains of each of eight pairs of guitarists playing a short melody together to explore the extent and the functional significance of synchronized cortical activity in the course of interpersonally coordinated actions. RESULTS: By applying synchronization algorithms to intra- and interbrain analyses, we found that phase synchronization both within and between brains increased significantly during the periods of (i) preparatory metronome tempo setting and (ii) coordinated play onset. Phase alignment extracted from within-brain dynamics was related to behavioral play onset asynchrony between guitarists. CONCLUSION: Our findings show that interpersonally coordinated actions are preceded and accompanied by between-brain oscillatory couplings. Presumably, these couplings reflect similarities in the temporal properties of the individuals' percepts and actions. Whether between-brain oscillatory couplings play a causal role in initiating and maintaining interpersonal action coordination needs to be clarified by further research. PMID- 19292893 TI - Breast cancer epidemiology according to recognized breast cancer risk factors in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial Cohort. AB - BACKGROUND: Multidisciplinary attempts to understand the etiology of breast cancer are expanding to increasingly include new potential markers of disease risk. Those efforts may have maximal scientific and practical influence if new findings are placed in context of the well-understood lifestyle and reproductive risk factors or existing risk prediction models for breast cancer. We therefore evaluated known risk factors for breast cancer in a cancer screening trial that does not have breast cancer as a study endpoint but is large enough to provide numerous analytic opportunities for breast cancer. METHODS: We evaluated risk factors for breast cancer (N = 2085) among 70,575 women who were randomized in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Using Poisson regression, we calculated adjusted relative risks [RRs, with 95% confidence intervals (CIs)] for lifestyle and reproductive factors during an average of 5 years of follow-up from date of randomization. RESULTS: As expected, increasing age, nulliparity, positive family history of breast cancer, and use of menopausal hormone therapy were positively associated with breast cancer. Later age at menarche (16 years or older vs. < 12: RR = 0.81, 95% CI, 0.65-1.02) or menopause (55 years or older vs. < 45: RR = 1.29, 95% CI, 1.03-1.62) were less strongly associated with breast cancer than was expected. There were weak positive associations between taller height and heavier weight, and only severe obesity [body mass index (BMI; kg/m(2)) 35 or more vs. 18.5-24.9: RR = 1.21, 95% CI, 1.02 1.43] was statistically significantly associated with breast cancer. CONCLUSION: The ongoing PLCO trial offers continued opportunities for new breast cancer investigations, but these analyses suggest that the associations between breast cancer and age at menarche, age at menopause, and obesity might be changing as the underlying demographics of these factors change. CLINICAL TRIALS REGISTRATION: (http://www.clinicaltrials.gov), NCT00002540. PMID- 19292894 TI - Measurements of left ventricular myocardial longitudinal systolic displacement using spectral and colour tissue Doppler: time for a reassessment? AB - BACKGROUND: Echocardiographic measurements of left ventricular (LV) myocardial displacement may produce different results depending on the choice of employed modality and subjective adjustments during data acquisition and analysis. METHODS: In this study, left ventricular longitudinal systolic displacement was quantified in 57 patients (31 women and 26 men, 50 +/- 16 years) using colour (colour TD) and spectral tissue Doppler (spectral TD) before and after temporal filtering (30 to 70 milliseconds in 20-millisecond steps) and changed offline gain saturation (0%, 50% and 100%), respectively. The results were compared with those obtained with anatomic M-mode. RESULTS: Whereas only minor differences occurred between the results of colour TD and anatomic M-mode measurements, spectral TD significantly overestimated the results obtained with both these methods. However, the limits of agreement between the results produced by all three studied methods were not clinically acceptable in any of the cases. The spectral TD displacement values increased along with increasing offline gain saturation whereas the effect of temporal filtering on colour Doppler measurements was insignificant. CONCLUSION: Measurements of LV myocardial longitudinal displacement employing spectral TD, colour TD or anatomic M-mode produce different results, thus discouraging interchangeable use of these modalities. Whereas the results of spectral TD measurements can be significantly altered by changing offline gain setting, the effect of temporal filtering on colour TD measurements is insignificant, a fact that increases clinical practicality of the latter method. PMID- 19292895 TI - Can performance indicators be used for pedagogic purposes in disaster medicine training? AB - BACKGROUND: Although disaster simulation trainings were widely used to test hospital disaster plans and train medical staff, the teaching performance of the instructors in disaster medicine training has never been evaluated. The aim of this study was to determine whether the performance indicators for measuring educational skill in disaster medicine training could indicate issues that needed improvement. METHODS: The educational skills of 15 groups attending disaster medicine instructor courses were evaluated using 13 measurable performance indicators. The results of each indicator were scored at 0, 1 or 2 according to the teaching performance. RESULTS: The total summed scores ranged from 17 to 26 with a mean of 22.67. Three indicators: 'Design', 'Goal' and 'Target group' received the maximum scores. Indicators concerning running exercises had significantly lower scores as compared to others. CONCLUSION: Performance indicators could point out the weakness area of instructors' educational skills. Performance indicators can be used effectively for pedagogic purposes. PMID- 19292896 TI - Statistical identification of gene association by CID in application of constructing ER regulatory network. AB - BACKGROUND: A variety of high-throughput techniques are now available for constructing comprehensive gene regulatory networks in systems biology. In this study, we report a new statistical approach for facilitating in silico inference of regulatory network structure. The new measure of association, coefficient of intrinsic dependence (CID), is model-free and can be applied to both continuous and categorical distributions. When given two variables X and Y, CID answers whether Y is dependent on X by examining the conditional distribution of Y given X. In this paper, we apply CID to analyze the regulatory relationships between transcription factors (TFs) (X) and their downstream genes (Y) based on clinical data. More specifically, we use estrogen receptor alpha (ERalpha) as the variable X, and the analyses are based on 48 clinical breast cancer gene expression arrays (48A). RESULTS: The analytical utility of CID was evaluated in comparison with four commonly used statistical methods, Galton-Pearson's correlation coefficient (GPCC), Student's t-test (STT), coefficient of determination (CoD), and mutual information (MI). When being compared to GPCC, CoD, and MI, CID reveals its preferential ability to discover the regulatory association where distribution of the mRNA expression levels on X and Y does not fit linear models. On the other hand, when CID is used to measure the association of a continuous variable (Y) against a discrete variable (X), it shows similar performance as compared to STT, and appears to outperform CoD and MI. In addition, this study established a two layer transcriptional regulatory network to exemplify the usage of CID, in combination with GPCC, in deciphering gene networks based on gene expression profiles from patient arrays. CONCLUSION: CID is shown to provide useful information for identifying associations between genes and transcription factors of interest in patient arrays. When coupled with the relationships detected by GPCC, the association predicted by CID are applicable to the construction of transcriptional regulatory networks. This study shows how information from different data sources and learning algorithms can be integrated to investigate whether relevant regulatory mechanisms identified in cell models can also be partially re-identified in clinical samples of breast cancers. AVAILABILITY: the implementation of CID in R codes can be freely downloaded from (http://homepage.ntu.edu.tw/~lyliu/BC/). PMID- 19292897 TI - Relationship between interRAI HC and the ICF: opportunity for operationalizing the ICF. AB - BACKGROUND: The International Classification of Functioning, Disability and Health (ICF) is embraced as a framework to conceptualize human functioning and disability. Health professionals choose measures to represent the domains of the framework. The ICF coding classification is an administrative system but multiple studies have linked diverse clinical assessments to ICF codes. InterRAI-HC (home care) is an assessment designed to assist planning of care for patients receiving home care. Examining the relationship between the ICF and the interRAI HC is of particular interest because the interRAI assessments are widely used in clinical practice and research, are computerized, and uploaded to databases that serve multiple purposes including public reporting of quality in Canada and internationally. The objective of this study was to examine the relationship between the interRAI HC (home care) assessment and the ICF. Specifically, the goal was to determine the proportion of interRAI HC items that can be linked to each of the major domains of the ICF (Body Function, Body Structure, Activities and Participation, and the Environmental Factors), the chapters and the specific ICF codes. METHODS: Three coders who were familiar with both the home care assessment and the ICF independently assigned ICF codes to inter-RAI HC items. Subsequently, a series of teleconference meetings were held to reach consensus on the primary code and much later consensus was used to finalize codes for additional items added to the interRAI HC. RESULTS: Following exclusion of administrative and diagnostic sections, 175 interRAI items were examined for potential assignment of codes. Of these 52 were assigned codes related to body function, 43 to activities and participation, 34 to environment, 1 to body structure, 17 to not coded, and 26 to not defined. Considering all 3-digit ICF codes, interRAI items addressed 43.2% of Body Function and 50.6% of Activities and Participation codes. CONCLUSION: The conceptual overlap in content, offers an excellent opportunity to operationalize the ICF domains and the codes particularly in the areas of Body Function and Activities and Participation. Use of measures such as the interRAI assessments with common elements across settings facilitates standardized reporting for organizations, regions and nations. PMID- 19292898 TI - Chromato-panning: an efficient new mode of identifying suitable ligands from phage display libraries. AB - BACKGROUND: Phage Display technology is a well established technique for high throughput screening of affinity ligands. Here we describe a new compact chromato panning procedure for selection of suitable binders from a phage peptide display library. RESULTS: Both phages and E. coli cells pass non-hindered through the interconnected pores of macroporous gel, so called cryogel. After coupling a ligand to a monolithic cryogel column, the phage library was applied on the column and non-bound phages were washed out. The selection of strong phage binders was achieved already after the first panning cycle due to the efficient separation of phage-binders from phage-non-binders in chromatographic mode rather than in batch mode as in traditional biopanning procedures. E. coli cells were applied on the column for infection with the specifically bound phages. CONCLUSION: Chromato-panning allows combining several steps of the panning procedure resulting in 4-8 fold decrease of total time needed for phage selection. PMID- 19292899 TI - The PsyCoLaus study: methodology and characteristics of the sample of a population-based survey on psychiatric disorders and their association with genetic and cardiovascular risk factors. AB - BACKGROUND: The Psychiatric arm of the population-based CoLaus study (PsyCoLaus) is designed to: 1) establish the prevalence of threshold and subthreshold psychiatric syndromes in the 35 to 66 year-old population of the city of Lausanne (Switzerland); 2) test the validity of postulated definitions for subthreshold mood and anxiety syndromes; 3) determine the associations between psychiatric disorders, personality traits and cardiovascular diseases (CVD), 4) identify genetic variants that can modify the risk for psychiatric disorders and determine whether genetic risk factors are shared between psychiatric disorders and CVD. This paper presents the method as well as sociodemographic and somatic characteristics of the sample. METHODS: All 35 to 66 year-old persons previously selected for the population-based CoLaus survey on risk factors for CVD were asked to participate in a substudy assessing psychiatric conditions. This investigation included the Diagnostic Interview for Genetic Studies to elicit diagnostic criteria for threshold disorders according to DSM-IV and algorithmically defined subthreshold syndromes. Complementary information was collected on potential risk and protective factors for psychiatric disorders, migraine and on the morbidity of first-degree relatives, whereas the collection of DNA and plasma samples was already part of the original CoLaus survey. RESULTS: A total of 3,691 individuals completed the psychiatric evaluation (67% participation). The gender distribution of the sample did not differ significantly from that of the general population in the same age range. Although the youngest 5-year band of the cohort was underrepresented and the oldest 5-year band overrepresented, participants of PsyCoLaus and individuals who refused to participate revealed comparable scores on the General Health Questionnaire, a self-rating instrument completed at the somatic exam. CONCLUSION: Despite limitations resulting from the relatively low participation in the context of a comprehensive and time-consuming investigation, the PsyCoLaus study should significantly contribute to the current understanding of psychiatric disorders and comorbid somatic conditions by: 1) establishing the clinical relevance of specific psychiatric syndromes below the DSM-IV threshold; 2) determining comorbidity between risk factors for CVD and psychiatric disorders; 3) assessing genetic variants associated with common psychiatric disorders and 4) identifying DNA markers shared between CVD and psychiatric disorders. PMID- 19292900 TI - Enhanced antitumor efficacy of cisplatin in combination with HemoHIM in tumor bearing mice. AB - BACKGROUND: Although cisplatin is one of the most effective chemotherapeutic agents, cisplatin alone does not achieve a satisfactory therapeutic outcome. Also cisplatin accumulation shows toxicity to normal tissues. In this study, we examined the possibility of HemoHIM both to enhance anticancer effect with cisplatin and to reduce the side effects of cisplatin in melanoma-bearing mice. METHODS: HemoHIM was prepared by adding the ethanol-insoluble fraction to the total water extract of a mixture of 3 edible herbs, Angelica Radix, Cnidium Rhizoma and Paeonia Radix. Anticancer effects of HemoHIM with cisplatin were evaluated in melanoma-bearing mice. We used a Cr51-release assay to measure the activity of NK/Tc cell and ELISA to evaluate the production of cytokines. RESULTS: In melanoma-bearing mice, cisplatin (4 mg/kg B.W.) reduced the size and weight of the solid tumors, and HemoHIM supplementation with cisplatin enhanced the decrease of both the tumor size (p < 0.1) and weight (p < 0.1). HemoHIM itself did not inhibit melanoma cell growth in vitro, and did not disturb the effects of cisplatin in vitro. However HemoHIM administration enhanced both NK cell and Tc cell activity in mice. Interestingly, HemoHIM increased the proportion of NK cells in the spleen. In melanoma-bearing mice treated with cisplatin, HemoHIM administration also increased the activity of NK cells and Tc cells and the IL-2 and IFN-gamma secretion from splenocytes, which seemed to contribute to the enhanced efficacy of cisplatin by HemoHIM. Also, HemoHIM reduced nephrotoxicity as seen by tubular cell of kidney destruction. CONCLUSION: HemoHIM may be a beneficial supplement during cisplatin chemotherapy for enhancing the anti-tumor efficacy and reducing the toxicity of cisplatin. PMID- 19292901 TI - Version control of pathway models using XML patches. AB - BACKGROUND: Computational modelling has become an important tool in understanding biological systems such as signalling pathways. With an increase in size complexity of models comes a need for techniques to manage model versions and their relationship to one another. Model version control for pathway models shares some of the features of software version control but has a number of differences that warrant a specific solution. RESULTS: We present a model version control method, along with a prototype implementation, based on XML patches. We show its application to the EGF/RAS/RAF pathway. CONCLUSION: Our method allows quick and convenient storage of a wide range of model variations and enables a thorough explanation of these variations. Trying to produce these results without such methods results in slow and cumbersome development that is prone to frustration and human error. PMID- 19292902 TI - Biomarkers of HIV-1 associated dementia: proteomic investigation of sera. AB - BACKGROUND: New, more sensitive and specific biomarkers are needed to support other means of clinical diagnosis of neurodegenerative disorders. Proteomics technology is widely used in discovering new biomarkers. There are several difficulties with in-depth analysis of human plasma/serum, including that there is no one proteomic platform that can offer complete identification of differences in proteomic profiles. Another set of problems is associated with heterogeneity of human samples in addition intrinsic variability associated with every step of proteomic investigation. Validation is the very last step of proteomic investigation and it is very often difficult to validate potential biomarker with desired sensitivity and specificity. Even though it may be possible to validate a differentially expressed protein, it may not necessarily prove to be a valid diagnostic biomarker. RESULTS: In the current study we report results of proteomic analysis of sera from HIV-infected individuals with or without cognitive impairment. Application of SELDI-TOF analysis followed by weak cation exchange chromatography and 1-dimensional electrophoresis led to discovery of gelsolin and prealbumin as differentially expressed proteins which were not detected in this cohort of samples when previously investigated by 2-dimensional electrophoresis with Difference Gel Electrophoresis technology. CONCLUSION: Validation using western-blot analysis led us to conclude that relative change of the levels of these proteins in one patient during a timeframe might be more informative, sensitive and specific than application of average level estimated based on an even larger cohort of patients. PMID- 19292903 TI - Understanding patients' perspective in the use of generic antiepileptic drugs: compelling lessons for physicians to improve physician/patient communication. AB - BACKGROUND: Epilepsy is a condition in which consistency of treatment is paramount to successful management and for most patients, effective seizure control can be achieved. Given the severe consequences of even a single breakthrough seizure, patients should be afforded every opportunity to succeed on their given regimens. DISCUSSION: Some experts argue that global policy on generic antiepileptic drug substitution in epilepsy should be limited - occurring at the discretion of and with careful monitoring by the physician. While the debate continues, physicians still have daily responsibilities to their patients to help them best manage their epilepsy within the context of the current environment - the reality of which may involve switching to a generic antiepileptic drug or navigating various formulations between generics. SUMMARY: To provide context, this paper first reviews the main "hot button" issues fueling the ongoing generic debate, including a broad overview of the current state of the literature. The main goal however is to provide physicians with a patient perspective on generic antiepileptic drug use in epilepsy as a source of clinically useful, everyday advice to improve communication and increase patient self-advocacy, both of which are necessary for optimal patient outcome. PMID- 19292904 TI - Construction and validation of a short-form quality-of-life scale for chinese patients with benign prostatic hyperplasia. AB - BACKGROUND: In 2003, a 74-item quality-of-life (QOL) scale for Chinese benign prostatic hyperplasia (BPH) patients (BPH-QLS) was developed. Although the scale displayed good reliability and validity, the time required to complete it may limit its use. The purpose of this study was to construct and validate a short form quality-of-life (QOL) scale for Chinese patients with Benign Prostatic Hyperplasia (BPH). METHODS: According to the previously published 74-item quality of life scale for BPH (BPH-QLS), we developed a pool of items, then condensed these items and validated the shortened scale, based on collected data from 163 patients with BPH. We used eight methods to reduce the items. RESULTS: A 33-item QOL scale for BPH (short-form of BPH-QLS) was constructed. The time take by the new scale was much shorter than the original one. There was no significant difference between the 33-item scale and 74-item scale, in terms of reliability. Moreover, the 33-item BPH-QLS showed a high correlation with the 74-item BPH-QLS (r = 0.971). Scores generated by the two scales were not only parallel and coincident but also at the same level. CONCLUSION: We concluded that the reliability and validity of the short form of BPH-QLS is close to those of the 74 item BPH-QLS. It should be a good choice in clinical practice for its greater compliance and clinical feasibility. PMID- 19292905 TI - Fostering participation of general practitioners in integrated health services networks: incentives, barriers, and guidelines. AB - BACKGROUND: While the active participation of general practitioners (GPs) in integrated health services networks (IHSNs) plays a critical role in their success, little is known about the incentives and barriers to their actual participation. METHODS: Data were gathered through semi-structured interviews and a mail survey with GPs enrolled in SIPA (system of integrated care for older persons) at 2 sites in Montreal. A total of 61 GPs completed the questionnaire, from which 22 were randomly selected for the qualitative study, with active and non-active participation in the IHSN. RESULTS: The key themes associated with GP participation were clinician characteristics, consequences perceived at the outset, the SIPA implementation process, relationships with the SIPA team and professional consequences. The incentive factors reported were collaborative practices, high rates of elderly and SIPA patients in their clienteles, concerns about SIPA, the selection of frail elderly patients, close relationships with the case manager, the perceived efficacy of SIPA, and improved professional practices. Barriers to GP participation included high expectations, GP recruitment, lack of information on SIPA, difficult relationships with SIPA geriatricians and deterioration of physician-patient relationships. Four profiles of participation were identified: 2 groups of participants active in SIPA and 2 groups of participants not active in SIPA. The active GPs were familiar with collaborative practices, had higher IHSN patient rates, expressed more concerns than expectations, reported satisfactory relationships with case managers and perceived the efficacy of SIPA. Both active and non-active GPs reported quality care in the IHSN and improved professional practice. CONCLUSION: Throughout the implementation process, the participation of GPs in an IHSN depends on numerous professional (clinician characteristics) and organizational factors (GP recruitment, relationships with case managers). Our study provides guiding principles for establishing future integrated models of care. It suggests practical guidelines to support the active participation of GPs in these networks such as physicians with collaborative practices, recruitment of significant number of patients per physicians, the information provided and the accompaniment by geriatricians. PMID- 19292906 TI - Egg banks in hypersaline lakes of the South-East Europe. AB - The cyst banks of 6 coastal hypersaline lakes of South-East Europe have been investigated. The study concerned the bottom sediments of Khersonesskoe and Koyashskoe lakes in the Crimea (Ukraine), Narte saltworks (Albania), Vecchia Salina at Torre Colimena (Apulia, Italy), Pantano Grande and Pantano Roveto at Vendicari (Sicily, Italy). A total of 19 cyst types were recognised. The cyst banks of lakes were found to be well separated in the representation derived from a statistical multivariate data analysis. For all the lakes examined a comparison was possible between the resting community in sediments (cyst bank) and the active one in the water. The cyst banks contained more species than those recorded over a multi-year sampling effort in the water column. The study of cyst hatching, performed on 5 cyst types under lab conditions, demonstrated that cysts do not hatch under the same conditions. Furthermore, each cyst type shows a wide range of preferential hatching conditions, which allow us to confirm the ecological generalism of salt lake species. PMID- 19292907 TI - Anorectal tuberculosis coexisting with adenocarcinoma: an unusual association. AB - INTRODUCTION: Tuberculosis affecting the anorectum is an uncommon extra-pulmonary form of the disease, and its association with malignancy is highly unusual. CASE REPORT: A 35 year lady presented with lower gastrointestinal bleed, altered bowel habit and significant weight loss. On examination, she had nodular stricture in the lower rectum, with friable mucosa, bleeding easily on touch. With the diagnosis of carcinoma lower rectum, she underwent abdomino-perineal resection of the growth. The histopathological examination revealed carcinoma rectum with coexisting tuberculosis. CONCLUSION: The aetiological association between the tuberculosis and anorectal cancer is a matter of debate. However, the treating surgeon should be aware of this association, to avoid confusion and delay in the management. PMID- 19292908 TI - Incidental carcinoma of the prostate gland presenting with initial manifestation of disseminated intravascular coagulopathy (dic) in a middle aged man: a case report. AB - BACKGROUND: Incidental carcinoma of the prostate gland is a common clinical problem among elderly males but this malignancy presenting initially with features of Disseminated Intravascular Coagulopathy (DIC) in the African blacks is rare. Disseminateded intravascular coagulathy is the most frequent coagulation disorder in patients with prostate cancer, However DIC as a first manifestation of prostate cancer is unusual. CASE REPORT: This paper reports a case of a 56 year old Nigerian civil servant who presented initially with clinical features of DIC characterised by bleeding from multilple orifices but was subsequently diagnosed at autopsy to be infiltrating adenocarcinoma of the prostate. CONCLUSION: This rare case of DIC should be considered especially in elderly men when no other cause can be found for coagulopathy. PMID- 19292909 TI - Atypical imaging findings in a renal transplant patient with reversible posterior leukoencephalopathy syndrome: a case report. AB - BACKGROUND: Atypical clinical and imaging findings in Reversible Posterior Leukoencephalopathy Syndrome are recognized with increasing frequency. CASE REPORT: We report a case of an adult in his 5(th )decade immunosupressed with methilprednisolone, tacrolimus and micophenolate who two months after renal transplantation, multiple infections and an episode of humoral rejection became hypertensive with severe headaches, visual field abnormalities, seizures, left hemiparesis and hemineglect. Computed Tomography scan of the brain showed a hypo dense lesion in the left occipital lobe. Ischemic stroke was diagnosed and aspirin and permissive hypertension were indicated. Twelve hours later he developed left sided motor seizures and cortical blindness. Magnetic Resonance Image showed hyper intensity in T2 and FLAIR in both occipital lobes and a small area of cortical restricted diffusion in Diffuson Weighted Images in the left occipital lobe. With a diagnosis of Reversible Posterior Leukoencephalopathy Syndrome his blood pressure was controlled with intravenous labetalol, and two days later the neurologic findings returned to baseline and most Computed tomography findings resolved. CONCLUSION: This case underscores that in the appropriate setting Reversible Posterior Leukoencephalopathy Syndrome should be suspected and the clinician should not be misled by atypical clinical or imaging findings. In contrast to other pathologies that resemble Reversible Posterior Leukoencephalopathy Syndrome, with the right and timely treatment, signs, symptoms and images can be completely reversible. PMID- 19292910 TI - Using non-invasive brain stimulation to augment motor training-induced plasticity. AB - Therapies for motor recovery after stroke or traumatic brain injury are still not satisfactory. To date the best approach seems to be the intensive physical therapy. However the results are limited and functional gains are often minimal. The goal of motor training is to minimize functional disability and optimize functional motor recovery. This is thought to be achieved by modulation of plastic changes in the brain. Therefore, adjunct interventions that can augment the response of the motor system to the behavioural training might be useful to enhance the therapy-induced recovery in neurological populations. In this context, noninvasive brain stimulation appears to be an interesting option as an add-on intervention to standard physical therapies. Two non-invasive methods of inducing electrical currents into the brain have proved to be promising for inducing long-lasting plastic changes in motor systems: transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (tDCS). These techniques represent powerful methods for priming cortical excitability for a subsequent motor task, demand, or stimulation. Thus, their mutual use can optimize the plastic changes induced by motor practice, leading to more remarkable and outlasting clinical gains in rehabilitation. In this review we discuss how these techniques can enhance the effects of a behavioural intervention and the clinical evidence to date. PMID- 19292911 TI - Mobility of primary health care workers in China. AB - BACKGROUND: Rural township health centres and urban community health centres play a crucial role in the delivery of primary health care in China. Over the past two and-a-half decades, these health institutions have not been as well developed as high-level hospitals. The limited availability and low qualifications of human resources in health are among the main challenges facing lower-level health facilities. This paper aims to analyse the mobility of health workers in township and community health centres. METHODS: Data used in this paper come from a nationwide survey of health facilities in 2006. Ten provinces in different locations and of varying levels of economic development were selected. From these provinces, 119 rural township health centres and 89 urban community health centres were selected to participate in a questionnaire survey. Thirty key informants were selected from these health facilities to be interviewed. RESULTS: In 2005, 8.1% and 8.9% of health workers left township and community health centres, respectively. The health workers in rural township health centres had three to 13 years of work experience and typically had received a formal medical education. The majority of the mobile health workers moved to higher-level health facilities; very few moved to other rural township health centres. The rates of workers leaving township and community health centres increased between 2000 and 2005, with the main reasons for leaving being low salaries, limited opportunities for professional development and poor living conditions. CONCLUSION: In China, primary health workers in township health centres and community health centres move to higher-level facilities due to low salaries, limited opportunities for promotion and poor living conditions. The government already has policies in place to counteract this migration, but it must step up enforcement if rural township health centres and urban community centres are to retain health professionals and recruit qualified health workers. PMID- 19292912 TI - Stereotactic radiosurgery for glioblastoma: retrospective analysis. AB - PURPOSE: This retrospective study was done to better understand the conditions for which stereotactic radiosurgery (SRS) for glioblastoma may be efficacious. METHODS: Between 2000 and 2007, 33 patients with a pathological diagnosis of glioblastoma received SRS with the Novalis Shaped Beam Radiosurgery system. Eighteen patients (54%) underwent salvage SRS for recurrence while 15 (45%) patients received upfront SRS following standard fractionated RT for newly diagnosed glioblastoma. RESULTS: There were no RTOG grade >2 acute side effects. The median survival after SRS was 6.7 months (range 1.4 - 74.7). There was no significant difference in overall survival (from the time of initial diagnosis) with respect to the timing of SRS (p = 0.2). There was significantly better progression free survival in patients treated with SRS as consolidation versus at the time of recurrence (p = 0.04). The majority of patients failed within or at the margin of the SRS treatment volume (21/26 evaluable for recurrence). CONCLUSION: SRS is well tolerated in the treatment of glioblastoma. As there was no difference in survival whether SRS is delivered upfront or at recurrence, the treatment for each patient should be individualized. Future studies are needed to identify patients most likely to respond to SRS. PMID- 19292914 TI - In silico prioritisation of candidate genes for prokaryotic gene function discovery: an application of phylogenetic profiles. AB - BACKGROUND: In silico candidate gene prioritisation (CGP) aids the discovery of gene functions by ranking genes according to an objective relevance score. While several CGP methods have been described for identifying human disease genes, corresponding methods for prokaryotic gene function discovery are lacking. Here we present two prokaryotic CGP methods, based on phylogenetic profiles, to assist with this task. RESULTS: Using gene occurrence patterns in sample genomes, we developed two CGP methods (statistical and inductive CGP) to assist with the discovery of bacterial gene functions. Statistical CGP exploits the differences in gene frequency against phenotypic groups, while inductive CGP applies supervised machine learning to identify gene occurrence pattern across genomes. Three rediscovery experiments were designed to evaluate the CGP frameworks. The first experiment attempted to rediscover peptidoglycan genes with 417 published genome sequences. Both CGP methods achieved best areas under receiver operating characteristic curve (AUC) of 0.911 in Escherichia coli K-12 (EC-K12) and 0.978 Streptococcus agalactiae 2603 (SA-2603) genomes, with an average improvement in precision of >3.2-fold and a maximum of >27-fold using statistical CGP. A median AUC of >0.95 could still be achieved with as few as 10 genome examples in each group of genome examples in the rediscovery of the peptidoglycan metabolism genes. In the second experiment, a maximum of 109-fold improvement in precision was achieved in the rediscovery of anaerobic fermentation genes in EC-K12. The last experiment attempted to rediscover genes from 31 metabolic pathways in SA 2603, where 14 pathways achieved AUC >0.9 and 28 pathways achieved AUC >0.8 with the best inductive CGP algorithms. CONCLUSION: Our results demonstrate that the two CGP methods can assist with the study of functionally uncategorised genomic regions and discovery of bacterial gene-function relationships. Our rediscovery experiments also provide a set of standard tasks against which future methods may be compared. PMID- 19292913 TI - RAGE (Receptor for Advanced Glycation Endproducts), RAGE ligands, and their role in cancer and inflammation. AB - The Receptor for Advanced Glycation Endproducts [RAGE] is an evolutionarily recent member of the immunoglobulin super-family, encoded in the Class III region of the major histocompatability complex. RAGE is highly expressed only in the lung at readily measurable levels but increases quickly at sites of inflammation, largely on inflammatory and epithelial cells. It is found either as a membrane bound or soluble protein that is markedly upregulated by stress in epithelial cells, thereby regulating their metabolism and enhancing their central barrier functionality. Activation and upregulation of RAGE by its ligands leads to enhanced survival. Perpetual signaling through RAGE-induced survival pathways in the setting of limited nutrients or oxygenation results in enhanced autophagy, diminished apoptosis, and (with ATP depletion) necrosis. This results in chronic inflammation and in many instances is the setting in which epithelial malignancies arise. RAGE and its isoforms sit in a pivotal role, regulating metabolism, inflammation, and epithelial survival in the setting of stress. Understanding the molecular structure and function of it and its ligands in the setting of inflammation is critically important in understanding the role of this receptor in tumor biology. PMID- 19292915 TI - Ancient diversification of eukaryotic MCM DNA replication proteins. AB - BACKGROUND: Yeast and animal cells require six mini-chromosome maintenance proteins (Mcm2-7) for pre-replication complex formation, DNA replication initiation and DNA synthesis. These six individual MCM proteins form distinct heterogeneous subunits within a hexamer which is believed to form the replicative helicase and which associates with the essential but non-homologous Mcm10 protein during DNA replication. In contrast Archaea generally only possess one MCM homologue which forms a homohexameric MCM helicase. In some eukaryotes Mcm8 and Mcm9 paralogues also appear to be involved in DNA replication although their exact roles are unclear. RESULTS: We used comparative genomics and phylogenetics to reconstruct the diversification of the eukaryotic Mcm2-9 gene family, demonstrating that Mcm2-9 were formed by seven gene duplication events before the last common ancestor of the eukaryotes. Mcm2-7 protein paralogues were present in all eukaryote genomes studied suggesting that no gene loss or functional replacements have been tolerated during the evolutionary diversification of eukaryotes. Mcm8 and 9 are widely distributed in eukaryotes and group together on the MCM phylogenetic tree to the exclusion of all other MCM paralogues suggesting co-ancestry. Mcm8 and Mcm9 are absent in some taxa, including Trichomonas and Giardia, and appear to have been secondarily lost in some fungi and some animals. The presence and absence of Mcm8 and 9 is concordant in all taxa sampled with the exception of Drosophila species. Mcm10 is present in most eukaryotes sampled but shows no concordant pattern of presence or absence with Mcm8 or 9. CONCLUSION: A multifaceted and heterogeneous Mcm2-7 hexamer evolved during the early evolution of the eukaryote cell in parallel with numerous other acquisitions in cell complexity and prior to the diversification of extant eukaryotes. The conservation of all six paralogues throughout the eukaryotes suggests that each Mcm2-7 hexamer component has an exclusive functional role, either by a combination of unique lock and key interactions between MCM hexamer subunits and/or by a range of novel side interactions. Mcm8 and 9 evolved early in eukaryote cell evolution and their pattern of presence or absence suggests that they may have linked functions. Mcm8 is highly divergent in all Drosophila species and may not provide a good model for Mcm8 in other eukaryotes. PMID- 19292916 TI - Decon2LS: An open-source software package for automated processing and visualization of high resolution mass spectrometry data. AB - BACKGROUND: Data generated from liquid chromatography coupled to high-resolution mass spectrometry (LC-MS)-based studies of a biological sample can contain large amounts of biologically significant information in the form of proteins, peptides, and metabolites. Interpreting this data involves inferring the masses and abundances of biomolecules injected into the instrument. Because of the inherent complexity of mass spectral patterns produced by these biomolecules, the analysis is significantly enhanced by using visualization capabilities to inspect and confirm results. In this paper we describe Decon2LS, an open-source software package for automated processing and visualization of high-resolution MS data. Drawing extensively on algorithms developed over the last ten years for ICR2LS, Decon2LS packages the algorithms as a rich set of modular, reusable processing classes for performing diverse functions such as reading raw data, routine peak finding, theoretical isotope distribution modelling, and deisotoping. Because the source code is openly available, these functionalities can now be used to build derivative applications in relatively fast manner. In addition, Decon2LS provides an extensive set of visualization tools, such as high performance chart controls. RESULTS: With a variety of options that include peak processing, deisotoping, isotope composition, etc, Decon2LS supports processing of multiple raw data formats. Deisotoping can be performed on an individual scan, an individual dataset, or on multiple datasets using batch processing. Other processing options include creating a two dimensional view of mass and liquid chromatography (LC) elution time features, generating spectrum files for tandem MS data, creating total intensity chromatograms, and visualizing theoretical peptide profiles. Application of Decon2LS to deisotope different datasets obtained across different instruments yielded a high number of features that can be used to identify and quantify peptides in the biological sample. CONCLUSION: Decon2LS is an efficient software package for discovering and visualizing features in proteomics studies that require automated interpretation of mass spectra. Besides being easy to use, fast, and reliable, Decon2LS is also open-source, which allows developers in the proteomics and bioinformatics communities to reuse and refine the algorithms to meet individual needs.Decon2LS source code, installer, and tutorials may be downloaded free of charge at http://http:/ncrr.pnl.gov/software/. PMID- 19292917 TI - Analysis of TNFAIP3, a feedback inhibitor of nuclear factor-kappaB and the neighbor intergenic 6q23 region in rheumatoid arthritis susceptibility. AB - INTRODUCTION: Genome-wide association studies of rheumatoid arthritis (RA) have identified an association of the disease with a 6q23 region devoid of genes. TNFAIP3, an RA candidate gene, flanks this region, and polymorphisms in both the TNFAIP3 gene and the intergenic region are associated with systemic lupus erythematosus. We hypothesized that there is a similar association with RA, including polymorphisms in TNFAIP3 and the intergenic region. METHODS: To test this hypothesis, we selected tag-single nucleotide polymorphisms (SNPs) in both loci. They were analyzed in 1,651 patients with RA and 1,619 control individuals of Spanish ancestry. RESULTS: Weak evidence of association was found both in the 6q23 intergenic region and in the TNFAIP3 locus. The rs582757 SNP and a common haplotype in the TNFAIP3 locus exhibited association with RA. In the intergenic region, two SNPs were associated, namely rs609438 and rs13207033. The latter was only associated in patients with anti-citrullinated peptide antibodies. Overall, statistical association was best explained by the interdependent contribution of SNPs from the two loci TNFAIP3 and the 6q23 intergenic region. CONCLUSIONS: Our data are consistent with the hypothesis that several RA genetic factors exist in the 6q23 region, including polymorphisms in the TNFAIP3 gene, like that previously described for systemic lupus erythematosus. PMID- 19292918 TI - Expression of TRPM8 in the distal cerebrospinal fluid-contacting neurons in the brain mesencephalon of rats. AB - BACKGROUND: It has been shown that distal cerebrospinal fluid-contacting neurons (dCSF-CNs) exist near the ventral midline of the midbrain aqueduct and also in the grey matter of the inferior third ventricle and the fourth ventricle floor in the superior segment of the pons. The dCSF-CNs communicate between the cerebrospinal fluid (CSF) and the brain parenchyma and may participate in the transduction and regulation of pain signals. The cold sensation receptor channel, TRPM8 is involved in analgesia for neuropathic pain, but whether the TRPM8 receptor exists on dCSF-CNs remains unknown. However, there is preliminary evidence that TRPM8 is expressed in dCSF-CNs and may participate in the transmission and regulation of sensory information between brain parenchyma and cerebrospinal fluid (CSF) in rats. METHODS: Retrograde tracing of the cholera toxin subunit B labeled with horseradish peroxidase (CB-HRP) injected into the lateral ventricle was used to identify dCSF-CNs. A double-labeled immunofluorescent technique and laser scanning confocal microscopy were used to identify the expression of TRPM8 in dCSF-CNs. Software Image-Pro Plus was used to count the number of neurons in three sections where CB-HRP positive neurons were located in the mesencephalon of six rats. RESULTS: The cell bodies of CB-HRP positive dCSF-CNs were found in the brain parenchyma near the midline of the ventral Aq, also in the grey of the 3V, and the 4V floor in the superior segment of the pons. In the mesencephalon their processes extended into the CSF. TRPM8 labeled neurons were also found in the same area as were CB-HRP/TRPM8 double labeled neurons. CB-HRP/TRPM8 double-labeled neurons were found in 42.9 +/- 2.3% of neurons labeled by TRPM8, and all CB-HRP-labeled neurons were also labeled with TPRM8. CONCLUSION: This study has demonstrated that the cold sensation receptor channel, TRPM8, is localised within the dCSF-CNs of the mesencephalon. TRPM8 acts as receptor of dCSF-CNs for sensation transmission and pain regulation. PMID- 19292919 TI - Early results of integrated malaria control and implications for the management of fever in under-five children at a peripheral health facility: a case study of Chongwe rural health centre in Zambia. AB - BACKGROUND: Zambia has taken lead in implementing integrated malaria control so as to attain the National Health Strategic Plan goal of "reducing malaria incidence by 75% and under-five mortality due to malaria by 20% by the year 2010". The strategic interventions include the use of long-lasting insecticide treated nets and indoor residual spraying, the use of artemisinin-based combination therapies (ACT) for the treatment of uncomplicated malaria, improving diagnostic capacity (both microscopy and rapid diagnostic tests), use of intermittent presumptive treatment for pregnant women, research, monitoring and evaluation, and behaviour change communication. Financial barriers to access have been removed by providing free malaria prevention and treatment services. METHODS: Data involving all under-five children reporting at the health facility in the first quarter of 2008 was evaluated prospectively. Malaria morbidity, causes of non-malaria fever, prescription patterns treatment patterns and referral cases were evaluated RESULTS: Malaria infection was found only in 0.7% (10/1378), 1.8% (251378) received anti-malarial treatment, no severe malaria cases and deaths occurred among the under-five children with fever during the three months of the study in the high malaria transmission season. 42.5% (586/1378) of the cases were acute respiratory infections (non-pneumonia), while 5.7% (79/1378) were pneumonia. Amoxicillin was the most prescribed antibiotic followed by septrin. CONCLUSION: Malaria related OPD visits have reduced at Chongwe rural health facility. The reduction in health facility malaria cases has led to an increase in diagnoses of respiratory infections. These findings have implications for the management of non-malaria fevers in children under the age of five years. PMID- 19292920 TI - Gamma-linolenic acid inhibits both tumour cell cycle progression and angiogenesis in the orthotopic C6 glioma model through changes in VEGF, Flt1, ERK1/2, MMP2, cyclin D1, pRb, p53 and p27 protein expression. AB - BACKGROUND: Gamma-linolenic acid is a known inhibitor of tumour cell proliferation and migration in both in vitro and in vivo conditions. The aim of the present study was to determine the mechanisms by which gamma-linolenic acid (GLA) osmotic pump infusion alters glioma cell proliferation, and whether it affects cell cycle control and angiogenesis in the C6 glioma in vivo. METHODS: Established C6 rat gliomas were treated for 14 days with 5 mM GLA in CSF or CSF alone. Tumour size was estimated, microvessel density (MVD) counted and protein and mRNA expression measured by immunohistochemistry, western blotting and RT PCR. RESULTS: GLA caused a significant decrease in tumour size (75 +/- 8.8%) and reduced MVD by 44 +/- 5.4%. These changes were associated with reduced expression of vascular endothelial growth factor (VEGF) (71 +/- 16%) and the VEGF receptor Flt1 (57 +/- 5.8%) but not Flk1. Expression of ERK1/2 was also reduced by 27 +/- 7.7% and 31 +/- 8.7% respectively. mRNA expression of matrix metalloproteinase-2 (MMP2) was reduced by 35 +/- 6.8% and zymography showed MMP2 proteolytic activity was reduced by 32 +/- 8.5%. GLA altered the expression of several proteins involved in cell cycle control. pRb protein expression was decreased (62 +/- 18%) while E2F1 remained unchanged. Cyclin D1 protein expression was increased by 42 +/- 12% in the presence of GLA. The cyclin dependent kinase inhibitors p21 and p27 responded differently to GLA, p27 expression was increased (27 +/- 7.3%) while p21 remained unchanged. The expression of p53 was increased (44 +/- 16%) by GLA. Finally, the BrdU incorporation studies found a significant inhibition (32 +/- 11%) of BrdU incorporation into the tumour in vivo. CONCLUSION: Overall the findings reported in the present study lend further support to the potential of GLA as an inhibitor of glioma cell proliferation in vivo and show it has direct effects upon cell cycle control and angiogenesis. These effects involve changes in protein expression of VEGF, Flt1, ERK1, ERK2, MMP2, Cyclin D1, pRb, p53 and p27. Combination therapy using drugs with other, complementary targets and GLA could lead to gains in treatment efficacy in this notoriously difficult to treat tumour. PMID- 19292921 TI - A novel approach to denoising ion trap tandem mass spectra. AB - BACKGROUND: Mass spectrometers can produce a large number of tandem mass spectra. They are unfortunately noise-contaminated. Noises can affect the quality of tandem mass spectra and thus increase the false positives and false negatives in the peptide identification. Therefore, it is appealing to develop an approach to denoising tandem mass spectra. RESULTS: We propose a novel approach to denoising tandem mass spectra. The proposed approach consists of two modules: spectral peak intensity adjustment and intensity local maximum extraction. In the spectral peak intensity adjustment module, we introduce five features to describe the quality of each peak. Based on these features, a score is calculated for each peak and is used to adjust its intensity. As a result, the intensity will be adjusted to a local maximum if a peak is a signal peak, and it will be decreased if the peak is a noisy one. The second module uses a morphological reconstruction filter to remove the peaks whose intensities are not the local maxima of the spectrum. Experiments have been conducted on two ion trap tandem mass spectral datasets: ISB and TOV. Experimental results show that our algorithm can remove about 69% of the peaks of a spectrum. At the same time, the number of spectra that can be identified by Mascot algorithm increases by 31.23% and 14.12% for the two tandem mass spectra datasets, respectively. CONCLUSION: The proposed denoising algorithm can be integrated into current popular peptide identification algorithms such as Mascot to improve the reliability of assigning peptides to spectra. AVAILABILITY OF THE SOFTWARE: The software created from this work is available upon request. PMID- 19292922 TI - Health risk behaviours among adolescents in the English-speaking Caribbean: a review. AB - BACKGROUND: The aim of this paper was to review and summarize research on prevalence of health risk behaviours, their outcomes as well as risk and protective factors among adolescents in the English-speaking Caribbean. METHODS: Searching of online databases and the World Wide Web as well as hand searching of the West Indian Medical Journal were conducted. Papers on research done on adolescents aged 10 - 19 years old and published during the period 1980 - 2005 were included. RESULTS: Ninety-five relevant papers were located. Five papers were published in the 1980s, 47 in the 1990s, and from 2000-2005, 43 papers. Health risk behaviours and outcomes were divided into seven themes. Prevalence data obtained for these, included lifetime prevalence of substance use: cigarettes-24% and marijuana-17%; high risk sexual behaviour: initiation of sexual activity 10 mg/dl). According to our laboratory data, elevated serum creatinine levels (> 1.3 mg/dL) indicated impaired renal function. RESULTS: In total, there were 5722 patients, including 2816 (49.2%) men and 2906 (50.8%) women, with a median age of 67. Impaired renal function was noted in 307 (5.4%) cases. Serum uric acid was significantly correlated with blood urea nitrogen and serum creatinine. Groups with a higher serum uric acid level had an increased risk of impaired renal function. CONCLUSION: Our purpose in this preliminary observation was to try to define a starting point for the early control of serum uric acid, in order to avoid the development of impaired renal function. We found that serum uric acid level to < 6 mg/dl seemed to be associated with less renal function impairment. PMID- 19292942 TI - Intraarterial Cisplatin and intravenous adriamycin in nonmetastatic osteosarcoma of the extremities: a single institution experience in Taiwan. AB - BACKGROUND: For nonmetastatic osteosarcoma of the extremities, the optimal treatment now consists of multiagent neoadjuvant and adjuvant chemotherapy and limb-sparing surgical procedures. The degree of tumor necrosis after neoadjuvant chemotherapy is one of the most important prognostic indicators. Intraarterial cisplatin and intravenous adriamycin could achieve a good initial tumor response and convert the response to an ultimate cure. METHODS: Between January 1989 and July 2004, 16 patients with nonmetastatic osteosarcoma of the extremities received intravenous adriamycin and intraarterial cisplatin monthly for 2-5 courses, based on achievement of a maximized angiographic response, followed by limb salvage surgery and then adjuvant intravenous chemotherapy with adriamycin and cisplatin. After resection, if patients had a good response (the extent of tumor necrosis was > or =90%), the same regimen was administered intravenously every three weeks for a total of six courses of chemotherapy. Poor responders (tumor necrosis <90%) were treated with a regimen of high-dose methotrexate with leucovorin rescue (HD-MTX) or ifosfamide, cisplatin, and etoposide (ICE). RESULTS: Patients received an average of four cycles of neoadjuvant intraarterial chemotherapy. Sixteen patients underwent limb-preservation surgery and 12 had >90% tumor necrosis. With an average follow-up of 93.5 months, 8 patients were continuously disease -free, 6 died of disease and 2 had no evidence of disease 112 and 182 months respectively after relapse. The 5-year overall survival rate was 61%. No patient developed clinically detectable cardiac toxicity or ototoxicity after adriamycin and cisplatin administration. Febrile neutropenia occurred infrequently. CONCLUSION: This study shows the effectiveness of treating nonmetastatic osteosarcoma of the extremities with intraarterial cisplatin and intravenous adriamycin infusion in Taiwan. However, the number of patients evaluated and treated in a single hospital was obviously too few to be considered statistically robust and this regimen deserves further testing in a multi institutional fashion. PMID- 19292943 TI - Outcome of the L5-S1 segment after posterior instrumented spinal surgery in degenerative lumbar diseases. AB - BACKGROUND: Posterior decompression, instrumentation, and posterolateral fusion are surgical procedures for the treatment of degenerative lumbar diseases. Solid fusion usually causes adjacent problems. This study investigated the clinical outcome and radiographic fate of the L5-S1 segment in patients who underwent posterior instrumented surgery for degenerative lumbar diseases. METHODS: From January 1999 to December 2000, 181 patients (average age 59.4 years, range 45-79 years) underwent posterior decompression, posterior instrumentation, and posterolateral fusion for degenerative lumbar diseases (including degenerative spondylolisthesis and degenerative lumbar scoliosis) with spinal stenosis. Modified Brodsky's criteria and the Oswestry disability index were used to evaluate patients before surgery and at the final followup. Degenerative changes in the L5-S1 intervertebral disc were evaluated with the University of California at Los Angeles (UCLA) grading scale. Adjacent L5-S1 segmental instability was defined as the appearance of retrolisthesis, anterolisthesis, or lateral listhesis in the static or dynamic radiographs at the final follow-up. RESULTS: Only 1 of these 181 patients developed inferior adjacent instability, but there were no symptoms related to this instability. The mean pre-operative L5-S1 disc degenerative score was 1.73 -/+ 0.66 and at the last follow-up, 1.87 -/+ 0.72 (p = 0.006). There was no symptomatic disc degeneration necessitating further L5-S1 fusion during follow-up. One hundred fifty-six patients (86%) exhibited satisfactory results (good or excellent). The mean Oswestry score was 21.8 -/+ 6.0 preoperatively, which improved to 9.6 -/+ 7.4 at the last follow-up (p = 0.001). CONCLUSIONS: The L5-S1 disc degenerated more after posterolateral lumbar floating fusion. However, there was no symptomatic inferior adjacent instability or symptomatic L5-S1 disc degeneration requiring further L5-S1 fusion at a mean 5.1 years follow-up. PMID- 19292944 TI - Results of microendoscopic discectomy performed in the 26 cases with a minimum 3 years follow-up. AB - BACKGROUND: Microendoscopic discectomy (MED) is less invasive than conventional open discectomy, but the long-term benefits of this technique are still debated. Controversy also remains regarding the surgical indications, patient selection, effectiveness, learning curve and complications. METHODS: From Dec 2001 to Dec 2003, 26 patients with lumbar herniated disc disease received MED. The surgical indications included the following: (1) unilateral, single level lumbar disc herniation; (2) signs and symptoms compatible with the involved nerve root; (3) failure of conservative treatment. These cases were the initial MEDs performed by one of our senior authors (TS FU). Clinical symptoms and outcomes were assessed using the Japanese Orthopaedic Association Back Scores. RESULTS: Treatment in two cases was changed to open discectomy because of irreparable dural tears during surgery. For the remaining 24 cases, the average intraoperative blood loss was 55.8 mL. The average operation length was 136.8 minutes and the average post surgical hospital stay was 2.4 days. At 12 weeks after the operation, 22 achieved excellent or good results. The satisfactory rate was 91.7%. On final follow-up, 21 patients had excellent or good results. The satisfactory rate was 87.5%. Complications included two irreparable dural tears, two superficial wound infections and one pseudomenigocele. CONCLUSIONS: Our data indicate that MED is an effective procedure for lumbar disc herniation. The result is satisfactory under adequate surgical indications and patient selection. Despite the low complication rate, dural tears still remain a concern during the learning stage. PMID- 19292945 TI - Thymic hyperplasia following successful chemotherapy for Hodgkin's lymphoma: report of a case. AB - A recurrent mediastinal mass in a patient following treatment for Hodgkin's lymphoma presents a diagnostic challenge. We report a 17-year-old boy with nodular sclerosing Hodgkin's lymphoma, stage IIIsA, who achieved complete response after 6-cycle chemotherapy with doxorubicin, bleomycin, vinblastine and dacarbazine and developed thymic hyperplasia 6 months later. Ten cases have been reported previously. Most patients were young, had the nodular sclerosis subtype of Hodgkin's lymphoma, were in an advanced stage, and had a latency of 2 to 12 months following successful chemotherapy. It is impossible to delineate radiologically benign from malignant lesions. Resection of the tumor for tissue diagnosis is mandatory to establish an accurate diagnosis and thus avoid further harmful therapy. PMID- 19292946 TI - Klatskin tumor with spermatic cord metastasis: a case report. AB - Malignant spermatic cord tumor is quite uncommon. Metastatic spermatic cord tumor is even rarer. Here we report a case of metastatic spermatic cord tumor from a common hepatic duct tumor (Klatskin tumor). A 38-year-old man presented with right scrotal enlargement and chronic testicular pain. He had a Klatskin tumor (cholangiocarcinoma) stage IIIa, and underwent an extended right hepatectomy with resection of the extrahepatic bile duct, and portal vein and reconstruction by Roux-en-y hepatico-jejunostomy one year before this presentation. Scrotal ultrasound revealed a moderate hydrocele with clean content. Scrotal exploration showed turbid fluid and a fibrotic hyperemic spermatic cord. Excision biopsy of the spermatic cord was done, and the pathology revealed adenocarcinoma. The primary cholangiocarcinoma tumor and the spermatic cord tumor showed identical histologic patterns. Klatskin tumor with spermatic cord metastasis was diagnosed. To our knowledge, this case represents the first such report in the literature. PMID- 19292947 TI - [Prenatal risk factors for neonatal asphyxia: how risk for each?]. AB - OBJECTIVE: Neonatal asphyxia is the third leading cause of neonatal death and main cause of long-term neurodevelopmental handicap throughout the world. Prevention is more important than treatment. Most previous reports are limited to retrospective investigations of the relationships between some prenatal risk factors and low Apgar scores. This study was designed to prospectively investigate the relationship between prenatal risk factors and neonatal asphyxia and the influence of single or multiple risk factors on the incidence of neonatal asphyxia, and examine significant risk factors for neonatal asphyxia. METHODS: From April 2002 through October 2004, a total of 10 376 live-born newborns were enrolled. Forty-six prenatal risk factors were investigated. Neonatal asphyxia was diagnosed based on the following four items: 1. 1-min Apgar score or=0.10 and p<0.05 as significant. The OR and 95%CI were calculated for each significant risk factor. RESULTS: Of the 10 376 newborns, 8 530 cases (82.21%) had 1-9 risk factors, and asphyxia occurred in 117 cases (1.13%) out of the 8 530 cases. In the 1 846 cases without risk factors, none had asphyxia (x2=25.6, p<0.01). The incidence of asphyxia increased with increasing numbers of risk factors, from 0.23% in newborns with one risk factor to 14.29% in newborns who had nine risk factors (r=0.96, p<0.01). Twelve significant risk factors identified were as follows: ominous fetal heart rate patterns (OR=17.1,95%CI:11.2 25.9), placenta abruption (OR=15.2, 95% CI: 4.5-51.8), maternal lung diseases (OR=11.5, 95% CI:1.4-91.3), fetal acidosis (OR=6.1, 95% CI:1.5-24.1), placenta previa (OR=5.0,95% CI:1.5-16.9), breech delivery (OR=4.5, 95% CI: 2.1-9.9), meconium stained amniotic fluid (OR=3.2, 95% CI:2.2-4.8), forcepsjassisted delivery (OR=3.2, 95%CI: 1.1-9.9), prolonged labor (OR=2.94, 95%CI:1.5-5.8), abnormal utero contraction (OR=2.8, 95% CI:1.7-4.6), and premature delivery (OR=2.5,95%CI:1.4-4.8). Cesarean section had a protective effect (OR=0.6, 95% CI:0.4-0.9) (all p<0.05). CONCLUSIONS: It is very important to prevent perinatal asphyxia by systematically examining prenatal risk factors and giving interventions for the newborns with risk factors, especially those with the above significant risk factors or with multiple risk factors. Proper cesareon section according to indications might be helpful to decrease the incidence of birth asphyxia. PMID- 19292948 TI - [Multicenter follow-up report of 147 premature infants with brain injuries from 6 hospitals in China]. AB - OBJECTIVE: Sponsored by the Subspecialty Group of Neonatology of Pediatric Society, China Medical Association, more than 10 large-scale hospitals participated in the near two-year multicenter investigation for Brain Injuries in Premature Infants in China. The present study presents the follow-up results of 147 premature infants with brain injuries from 6 Third Class A Level hospitals. METHODS: All premature infants with intraventricular hemorrhage (IVH) or periventricular leukomalacia (PVL) diagnosed in the early neonatal period in the 6 hospitals were followed-up between January 2005 and August 2006. Based on the synthetic results of physical development, examination of nervous system, intelligence tests and cranial ultrasound, the premature infants with brain injuries were classified as normal development, marginal development and retarded development. RESULTS: One hundred and forty-seven premature infants with brain injuries from the 6 hospitals consisted of 141 cases of IVH and 36 cases of PVL (30 cases having IVH and PVL). Based on the synthetic follow-up results, 51.4% of premature infants with brain injuries were generally assessed as normal development, 38.4% as marginal development and 10.7% as retarded development. Among them, delayed growth in head circumference, height and weight was 13.4%; the occurrence frequency of cerebral paralysis (CP) was 7.1% in PVL grade I, 28.6% in PVL grade II and 100% in PVL grade III; 12.7% showed retarded development of intelligence; and 30% presented post-injurious changes on cranial sonography. CONCLUSIONS: The data of the multicenter follow-up can basically reflect the short-term prognosis of premature infants with brain injuries in major big cities of China. About 10% of them have retarded physical, motor-and mental developments. The long-term regular follow-up study is expected for more premature infants with brain injuries, and behavioral sequelae of brain injuries which may occur in peri-school age and adolescence should be paid particularly close attention. PMID- 19292949 TI - [Changes of electroencephalographic background patterns and serum neuron specific enolase levels in neonates with hypoxic-ischemic encephalopathy]. AB - OBJECTIVE: To examine electroencephalographic (EEG) background patterns and serum neuron specific enolase (NSE) levels in neonates with hypoxic-ischemic encephalopathy (HIE) in order to study their roles in assessing clinical progress, severity and prognosis in children with HIE. METHODS: A total of 58 neonates with HIE, including 38 cases of mild, 12 cases of moderate and 8 cases of severe HIE, were enrolled. Thirty normal neonates were used as the control group. Serum NES levels were measured by radioimmunoassay 12-24 hrs and 7-10 days after birth. EEG examination was performed 24 hrs-18 days of age (early stage) and 28-30 days of age (convalescence stage). The neonates with HIE were followed up, with a duration of 6 months to 3 years. RESULTS: Fifty-five neonates with HIE (94.8%) showed EEG abnormalities in the early stage, but only 2 patients (6.7%) in the control group (p<0.01). EEG background patterns abnormalities in the early stage were found in 90.0% (18/20) of moderate-severe HIE neonates. The proportion was significantly higher than that in the mild HIE group [5.2% (2/38); p<0.01]. Thirteen (72.2%) out of 18 patients with early EEG background patterns abnormalities had poor outcomes. Serum levels of NES in the HIE group were significantly higher than those in the control group 12-24 hrs after birth (p<0.01). EEG background patterns abnormalities and increased serum levels of NES 12-24 hrs after birth were consistent with the clinical grading of HIE. Most of neonates [87.5% (7/8)] who showed abnormal EEG background patterns at the convalescence stage had neurological sequelae. CONCLUSIONS: EEG background patterns and serum NSE levels may be useful in assessment of disease severity and neurological outcome in children with HIE. PMID- 19292950 TI - [Effect of impaired glucose tolerance during pregnancy on newborns]. AB - OBJECTIVE: Abnormal glucose metabolism during pregnancy can result in significant adverse outcomes for newborns. This study was designed to investigate the effect of different degrees of impaired glucose tolerance on newborns. METHODS: Glucose challenge test (GCT) and oral glucose tolerance test (OGTT) were performed in pregnant women at 24-28 weeks of gestation. Based on the results of the two tests, their newborn infants were classified into five groups: gestational diabetes mellitus (GDM, n=182), gestational impaired oral glucose tolerance [GIGT, subdividing into GIGT 1 h (n=57) and GIGT 2-3 h groups (n=156)based on the occurrence time of abnormal blood glucose after glucose load], GCT abnormal but OGTT normal (only GCT abnormal, n=38) and normal glucose tolerance (control, n=1 025). The perinatal outcomes were compared among the five groups. RESULTS: The incidences of macrosomia, large for gestational age (LGA), small for gestational age (SGA), neonatal hypoglycemia and premature birth in the GIGT (1 h) group were significantly higher than those in the control group, but similar to those in the GDM group. The incidences of macrosomia, SGA, neonatal hypoglycemia and premature birth in the GIGT (2-3 h) and the only GCT abnormal groups significantly decreased as compared with those in the GDM group, and similar to those in the control group. GIGT (1 h ) group showed higher incidence of neonatal hypoglycemia and premature birth than the GIGT (2-3 h) and the only GCT abnormal groups. CONCLUSIONS: Different degrees of maternal impaired glucose tolerance have different effects on newborns. As maternal GDM, maternal GIGT (1 h) may increase the risk of macrosomia, LGA, SGA, neonatal hypoglycemia and premature birth. PMID- 19292951 TI - [Differentiation of hypoxic-ischemic encephalopathy and acute bilirubin encephalopathy with magnetic resonance imaging in neonates]. AB - OBJECTIVE: To investigate the value of conventional magnetic resonance imaging (MRI) and diffusion weighed imaging (DWI) in the differentiation of hypoxic ischemic encephalopathy (HIE) and acute bilirubin encephalopathy in neonates. METHODS: The MRI findings along with DWI characteristics in 15 neonates with HIE involving basal ganglia and in 18 neonates with acute bilirubin encephalopathy between November 2006 and June 2008 were retrospectively reviewed. RESULTS: On T1WI, only 5 patients presented hyperintensity in the globus pallidus in the HIE group, but 16 in the acute bilirubin encephalopathy group (p<0.01). Nine patients in the HIE group showed hyperintensity in the putamen, but the hyperintensity in the putamen was not found in the acute bilirubin encephalopathy group. The frequency of hyperintensity in the subthalamus in the acute bilirubin encephalopathy group (55.6%) was significantly higher than that in the HIE group (13.3%) (p<0.05). Eight patients in the HIE group showed abnormal signals in the other regions on T1WI, but only two patients in the acute bilirubin encephalopathy group (p<0.05). On DWI, 7 out of 11 patients with HIE presented hyperintensity in the basal ganglia, while all 10 patients of the acute bilirubin encephalopathy group presented normal in the basal ganglia. CONCLUSIONS: Conventional MRI along with DWI is useful in differentiating HIE from acute bilirubin encephalopathy in neonates. PMID- 19292952 TI - [Clinical significance of anti-D IgG screening and titer detection in 286 RhD negative pregnant women]. AB - OBJECTIVE: Anti-D IgG in RhD negative pregnant women is the main antibody of Rh induced hemolytic disease of newborn (HDN). The study aimed to investigate the clinical significance of anti-D IgG screening and titer detection in RhD negative pregnant women. METHODS: Sera of 286 RhD negative pregnant women were collected. Microtube column indirect antiglobulin test was used to screen and identify anti D IgG. The indirect antiglobulin test was used to test the titer of anti-D IgG. RESULTS: Anti-D IgG was identified in 21 cases (7.3%). The titer of anti-D showed an increasing trend with pregnancy progresses. The clinical outcomes of 12 fetuses (newborns) from positive anti-D pregnant women were observed. Two cases died in utero, 2 cases did not show abnormality and 8 cases had hemolysis. The 8 cases with hemolysis were treated with exchange transfusion or blood transfusion, and they had a good prognosis. CONCLUSIONS: The screening and titer detection of anti-D IgG in RhD negative pregnant women are valuable in the prediction and treatment of HDN. PMID- 19292953 TI - Evaluation of the effects of surfactant replacement therapy in neonatal respiratory distress syndrome. AB - OBJECTIVE: Respiratory distress syndrome (RDS) is a major cause of morbidity and mortality in preterm neonates. Pulmonary surfactant deficiency is the primary cause of RDS. The purpose of this study was to determine the effect of surfactant therapy in reduction of the mortality rate in premature neonates with RDS and to assess the relationship between the efficacy of surfactant therapy and some risk factors associated with RDS. METHODS: This study comprised 89 premature neonates with signs of RDS. The neonates were selected by simple sampling from those admitted to the Neonatal Intensive Care Unit (NICU) of Shaheed Beheshti Hospital. The eligible neonates received surfactant replacement-therapy (100 mg/kg) during 48 hours after birth. RESULTS: Overall, 34 (38.2%) out of 89 neonates who received surfactant survived. The higher efficacy of surfactant replacement therapy was observed in neonates with gestational age of more than 32 weeks (47.5%), in those who received the first dose of surfactant during the first 24 hours of life (43.3%), in those with an Apgar score of more than 7/10 at 1 and 5 min (48.1%), and in those with a birth weight of more than 1 500 g (52.5%). The neonates whose mother received steroid therapy before labor had higher reduction in mortality after surfactant therapy (41.7% with steroid vs 34.2% without steroid; p<0.05). CONCLUSIONS: Surfactant replacement therapy in neonatal RDS should be started as soon as possible after birth. It could reduce the mortality rate from RDS by 38.2%. The efficacy of surfactant therapy for neonatal RDS may be associated with gestational age, Apgar score, birth weight, starting time of surfactant therapy and maternal steroid therapy. PMID- 19292954 TI - [Neurobehavioral function of neonatal mice following excitotoxic brain damage]. AB - OBJECTIVE: To assess the changes of neurobehavioral function in a neonatal mouse model of excitotoxic brain damage. METHODS: Fifty-five 5-day-old ICR neonatal mice were randomly assigned to three groups: blank (no intravenous) control (n=20), saline control (n=20) and excitotoxic brain damage model (ibotenic acid treatment, n=15). Behavioral function was evaluated by the surface righting reflex test (postnatal days 6-10), the swimming test (postnatal days 8-12) and the Y-maze discrimination learning test (postnatal days 33-34). RESULTS: Righting time in the surface righting reflex test in the ibotenic acid treatment group on postnatal days 6-10 was more prolonged than that in the two control groups (p<0.05). Swimming test scores in the ibotenic acid treatment group were significantly lower than those in the two control groups (p<0.05). In the Y-maze discrimination learning test, the mice from the ibotenic acid treatment group performed significantly worse than two control groups, presenting with increased learning times (19.79+/-2.42 vs 16.29+/-2.48 or 16.30+/-2.37; p<0.05) and achieving a lower correct percentage (86.7% vs 96.5% or 95.0%) (p<0.05). CONCLUSIONS: The developmental reflexes and learning and memory functions were impaired in neonatal mice following excitotoxic brain damage. Behavioral testing is useful in the evaluation of early developmental reflexes and long-term neurobehavioral outcome in neonatal mice with excitotoxic brain damage. PMID- 19292955 TI - [Anomalous origin of the right pulmonary artery in a neonate]. PMID- 19292956 TI - [A case report of severe neonatal haemolysis due to mother and her newborn having the same blood type of Rh weak D]. PMID- 19292957 TI - [A case report of neonatal polycystic kidney]. PMID- 19292958 TI - [Congenital analgesia: a case report and literature review]. PMID- 19292959 TI - [Changes of prostaglandin D2 receptor on T cells in peripheral blood of children with asthma]. AB - OBJECTIVE: Chronic airway inflammation is associated with the polarization of TH2 cells in asthma. Prostaglandin D2 (PGD2) plays an important role in the polarization of TH2 cells. This study aimed to investigate the changes of PGD2 receptors (DP1/CRTH2) on T lymphocytes and their significance in asthma. METHODS: Seventy-two children with asthma were assigned to two groups: acute attack (n=42) and remission (n=30). Thirty-five healthy children were used as the control group. Plasma levels of TH2 cytokines IL-4 and IL-5, and TH1 cytokine INF-gamma were detected using ELISA. Radiological binding assay (RBA) was used to measure the contents of DP1/CRTH2 receptors on T cells in peripheral blood (PPB). RESULTS: The total combining contents of DP and CRTH2 on T cells in PPB in the acute attack and the remission groups were significantly higher than those in the control group (p<0.01). There was no significant difference in the DP1 content among the three groups. Serum levels of IL-4 and IL-5 significantly increased (p<0.01), in contrast, serum levels of TH1 cytokine IFN-gamma were significantly reduced in the acute attack and the remission groups compared with those in the control group (p<0.01). CONCLUSIONS: The total combining contents of DP and CRTH2 on T cells increased, serum levels of TH2 cytokines also increased, but serum levels of TH1 cytokine decreased significantly in the acute attack and the remission phases in children with asthma. This showed that a polarization of TH2 cells occurred in children with asthma and suggested that CRTH2 antagonism may be a new target for the treatment of asthma. PMID- 19292960 TI - [Viral pathogens of acute lower respiratory tract infection in hospitalized children from East Guangdong of China]. AB - OBJECTIVE: To investigate the viral pathogens of acute lower respiratory tract infection (ALRTI) in hospitalized children from East Guangdong Province of China and the relationship of the pathogens with age and seasons. METHODS: The nasopharyngeal aspirates samples obtained from 345 hospitalized children with ALRTI were investigated for respiratory syncytial virus (RSV), human bocavirus (HBoV), human metapneumovirus (hMPV), influenza virus types A and B, rhinovirus, parainfluenza virus types 1 and 3 and adenovirus by PCR. RESULTS: Viral pathogens were detected in 178 patients (51.6%). RSV was the most frequent (19.3%). Novel viruses hMPV (3.2%) and HBoV (3.2%) were found. A highest detection rate (61.9%) of virus was found between January to March. The infants aged 1 to 6 months showed a higher detection rate (71.3%) of virus than the other age groups. The detection rate of viral pathogens was 72.6% in children with bronchiolitis, followed by asthmatic bronchitis (70.0%) and bronchial pneumonia (44.6%). CONCLUSIONS: RSV remained the leading viral pathogens in children with ALRTI in East Guangdong of China. Novel viruses HBoV and hMPV were also important pathogens. The detection rate of viral pathogens was associated with seasonal changes and age. Different respiratory infectious diseases had different viral detection rates, with highest detection rate in bronchiolitis cases. PMID- 19292961 TI - [Effect of conductive education combined with Frenkel training on balance disability in children with cerebral palsy]. AB - OBJECTIVE: To study the efficacy of conductive education combined with Frenkel training in the improvement of balance function in children with cerebral palsy. METHODS: One hundred and fifteen children with cerebral palsy were randomly administered with conductive education and Frenkel training (study group, n=60) or conventional training (control group, n=55). Activities of daily living (ADL) scale and gross motor function measurement (GMFM) of physical performances were used to assess the balance function. RESULTS: The scores of ADL scale and GMFM of physical performances in both the study and the control groups increased after training. The study group showed higher scores of ADL scale (37.91+/-10.12 vs 34.18+/-6.13; p<0.05)and GMFM (62.93+/-15.00 vs 54.53+/-14.11) than the control group (p<0.05). CONCLUSIONS: Conductive education combined with Frenkel training is more effective for the improvement of balance function in children cerebral palsy. PMID- 19292962 TI - [Antimicrobial resistance of 36 strains of Helicobacter pylori in adolescents]. AB - OBJECTIVE: To investigate the resistance of Helicobacter pylori (H. pylori) to antimicrobial agents in adolescents. METHODS: One hundred and eight adolescents (6-18 years old) underwent a gastroscopy examination. H. pylori strains were isolated from gastric mucosa. Antimicrobial susceptibility testing was performed by means of Kirby-Baner. RESULTS: Thirty-six H. pylori strains were identified. The resistant rate of H. pylori strains to clarithromycin, fruranzolidone and amoxicillin was 8.3%, 16.7% and 33.3%, respectively. H. pylori strains showed a high resistance to metronidazole (94.4%), while no strain was resistant to gentamicin and levofloxacin. CONCLUSIONS: The H. pylori strains from the adolescents in Zhejiang showed a high resistance to metronidazole and amoxicillin but a low resistance to clarithromycin and fruranzolidone. All of H. pylori strains were susceptive to gentamicin and levofloxacin. PMID- 19292963 TI - [Distribution of connexin 43 and interstitial cells of Cajal in bowels of children with Hirschsprung's disease]. AB - OBJECTIVE: To investigate the distribution of connexin 43 (Cx43) and interstitial cells of Cajal (ICC) in bowels of children with Hirschsprung's disease (HD) and explore their roles in the pathogenesis of HD. METHODS: Forty-two children with HD diagnosed by histopathology (33 males and 9 females) aged from 2 months to 10 years were enrolled. Expression of Cx43 and ICC in the bowels was detected using immunohistochemistry. These cases were all sporadic, including 30 cases of common type and 12 cases of short segment type. Five samples from the children with intussusception (aged from 1 month to 8 years) were used as controls. RESULTS: Cx43 and ICC were not expressed in the muscle layers of the aganglionic segment in children with HD, which was significantly different from the Cx43 and ICC expression in the ganglionic segment of HD patients and the control bowels. Moderate expression of Cx43 and ICC were observed in the migratory segment, which was significantly different from that in the ganglionic and aganglionic segments in patients with HD. Moderate or strong expression of Cx43 and ICC was observed in the circular muscle and the region between the circular and longitudinal layer in the ganglionic segment from patients with HD. There were no significant differences in the Cx43 and ICC expression between the ganglionic segment of HD patients and control bowels. CONCLUSIONS: The absence of Cx43 and ICC expression in the aganglionic bowel and the destruction of the gap junction may induce the dysfunctions of intercellular substance exchange and communication transmission, which might partly be responsible for the pathogenesis of HD. PMID- 19292964 TI - [Serotypes and ampicillin resistance of Haemophilus influenzae isolates from children with respiratory infection in Hangzhou]. AB - OBJECTIVE: To investigate the serotypes distribution and ampicillin resistance of Haemophilus influenzae isolates from children with respiratory infection in Hangzhou. METHODS: Haemophilus influenzae strains were identified with V factor and X factor tests. Serotypes were determined with the slide agglutination method. Nitrocefin test was used to detect beta-lactamase. The sensitivities of ampicillin to Haemophilus influenzae were determined with the Kirby-Bauer diffusion method and the E-test method. RESULTS: One hundred and fifty-two Haemophilus influenzae isolates, 108 from boys and 44 from girls, were identified between December 2006 and July 2007. Of the 152 isolates, 148 (97.4%) were untypable, only 4 (2.6%) were typable, including type a, type d, type e and type f (n=1 each type). Haemophilus influenzae type b and c strain was not found. Thirty-four isolates (22.4%) were beta-lactamase-positive. One hundred and thirteen isolates (74.3%) were susceptible to ampicillin, while 34 isolates (22.4%) were resistant to ampicillin. CONCLUSIONS: Untypable strains were the most common in Haemophilus influenzae isolates from children with respiratory infection in Hangzhou. The isolates of Haemophilus influenzae kept susceptibity to ampicillin to a certain extent. PMID- 19292965 TI - [Perturbed hepatic phosphoinositol 3-kinase signaling pathway in the rat with intrauterine growth restriction]. AB - OBJECTIVE: To determine the molecular mechanisms linking intrauterine growth restriction (IUGR) to adult type 2 diabetes mellitus, the effect of IUGR on the hepatic post-receptor insulin-signaling pathway was investigated in the adult offspring. METHODS: The IUGR model was prepared by maternal protein-malnutrition. Western blotting analysis was undertaken to assess hepatic expression of insulin receptor substrate (IRS-2), phosphoinositol 3-kinase (PI-3K), protein kinase B (PKB), phosphorylated PKB-Ser473 and glycogen synthase kinase (GSK) 3 in 8-week old male IUGR rats. RESULTS: The basal levels of PI-3K protein decreased in IUGR rats compared with normal controls (p<0.01), whereas GSK-3beta protein level significantly increased in IUGR rats (p<0.01). Both PKB and phosphorylated PKB Ser473 protein levels significantly decreased in the liver of IUGR rats compared with normal controls (p<0.01)). After insulin administration, phosphorylated PKB Ser473 significantly increased to 182% of basal level in control rats(p<0.01); However, phosphorylation of PKB which responded to insulin was markedly blunted in IUGR rats compared with controls and only increased to 123% of basal level (p<0.05). CONCLUSIONS: The level of PI-3K and PKB and phosphorylated PKB-Ser473 expression decreased in the liver of IUGR rats, whereas the levels of GSK-3beta protein increased. It may contribute to the pathogenesis of insulin resistance in the IUGR rats. PMID- 19292966 TI - [Comorbidities and behavioral problems in children with functional articulation disorder]. AB - OBJECTIVE: To study the incidences of comorbidities and behavioral problems in children with functional articulation disorders. METHODS: One hundred and twelve children with functional articulation disorders (aged 4-11 years) were enrolled. Their comorbidities were identified based on clinical investigations and the DSM IV diagnosis criteria of attention deficit hyperactivity disorder (ADHD), stuttering, tic disorders and enuresis. Behavioral problems were evaluated by the Conners Parent Symptom Questionnaire and the Child Behavior Checklist. RESULTS: Sixty-nine patients (61.6%) had one or more comorbidities. The incidence of comorbidity in children with middle-severe functional articulation disorders was higher than in those with mild disorders. The most common comorbidity was language impairment (30.4%), followed by stuttering (16.1%), enuresis (13.4%), and tic disorders (6.3%). In school age children, ADHD (47.5%) was the most common comorbidity. The incidence of behavioral problems was 40.2% by the Child Behavior Checklist and 57.1% by the Parent Symptom Questionnaire. CONCLUSIONS: The children with functional articulation disorders have high incidence of comorbidity and many behavioral problems. PMID- 19292967 TI - [Efficacy of fosinopril on proteinuria in children with Henoch-Schonlein purpura nephritis]. PMID- 19292968 TI - [Problems and solutions in the diagnosis and therapy of hand-foot-mouth disease]. PMID- 19292969 TI - [Primary Sjogren's syndrome in children: a case report of 4 case]. PMID- 19292970 TI - [Rett syndrome in a child treated with adrenocorticotrophic hormone]. PMID- 19292971 TI - [Oxidants/antioxidants imbalance and hyperoxia-induced lung injury]. PMID- 19292972 TI - [Current application of glucocoriticoid in juvenile idiopathic arthritis]. PMID- 19292973 TI - Proceedings and data from The Schizophrenia Summit: a critical appraisal to improve the management of Schizophrenia. AB - In mid-February 2008, a panel of 10 nationally recognized schizophrenia research experts was assembled in a Schizophrenia Summit to focus on controversies that exist in treating patients with schizophrenia. The current literature related to the diagnosis and etiopathology of schizophrenia was evaluated regarding the identification of a prodromal phase, brain changes, cognitive impairments, genetic factors, and use of neuroimaging in patients with schizophrenia. Further, consideration was given to evidence supporting the neuroprotective benefits of atypical antipsychotic medications, the benefits of treating patients during the prodromal period, the use of combination antipsychotic medications, the need to improve cognitive function, and the management of substance abuse. Summit faculty member opinion is compared with field survey results, and recommendations are made for future research. PMID- 19292974 TI - WITHDRAWN: Modulation of microRNA processing by p53. AB - This article has been withdrawn at the request of the editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19292975 TI - Presenilin 1 modifies lipid raft composition of neuronal membranes. AB - Protein-lipid interactions in the nervous system may provide insight into the causes of neurological disorders. In this study, we elucidated if expression of human presenilin 1 (PS1) in a mouse model changes the physico-chemical properties of brain membranes. PS1 is a multifunctional transmembrane protein and part of the gamma-secretase complex. This complex is critical for the production of the Alzheimer related amyloid beta peptide. Brain membranes isolated from mice expressing a human wild-type PS1 transgene are less fluid and contain higher cholesterol and sphingomyelin levels. Moreover, our data reveal significant changes in membrane micro-domains and indicate that PS1 induces the formation of lipid rafts. PMID- 19292976 TI - Inhibition of LPS-induced pulmonary inflammation by specific flavonoids. AB - In the present study, the anti-inflammatory effects of the flavonoids flavone, fisetin and tricetin were evaluated in a mouse model of LPS-induced acute pulmonary inflammation. The flavonoid fisetin significantly reduced lung myeloperoxidase-levels and gene-expression of inflammatory mediators such as IL 6, TNF-alpha, IL-1beta, MIP-1alpha and MIP-2. The LPS-induced gene transcription of HO-1 and SOD2 was also significantly reduced by fisetin. Overall, the anti inflammatory effects of fisetin in this in vivo model were much more pronounced as compared to the observed effects of flavone or tricetin and the anti inflammatory glucocorticoid dexamethasone. The results of this study indicate that flavonoids such as fisetin might be potential candidates as pharmaceuticals or nutraceuticals in the treatment of pulmonary inflammatory diseases. PMID- 19292977 TI - Acute pancreatitis markedly accelerates pancreatic cancer progression in mice expressing oncogenic Kras. AB - Chronic pancreatitis increases by 16-fold the risk of developing pancreatic ductal adenocarcinoma (PDAC), one of the deadliest human cancers. It also appears to accelerate cancer progression in genetically engineered mouse models. We now report that in a mouse model where oncogenic Kras is activated in all pancreatic cell types, two brief episodes of acute pancreatitis caused rapid PanIN progression and accelerated pancreatic cancer development. Thus, a brief inflammatory insult to the pancreas, when occurring in the context of oncogenic Kras(G12D), can initiate a cascade of events that dramatically enhances the risk for pancreatic malignant transformation. PMID- 19292978 TI - COX-2 as a novel target of CRF family peptides' participating in inflammation. AB - In mammals, corticotropin-releasing factor (CRF) family peptides include CRF, Urocortin (Ucn) 1, Ucn2, and Ucn3. In contrast to their systemic indirect immunosuppressive effects on the hypothalamic-pituitary adrenal axis, CRF family peptides act as locally expressed autocrine or paracrine pro-inflammatory factors in a series of inflammatory diseases. Cyclooxygenase-2 (COX-2), the rate-limiting enzyme in metabolism of arachidonic acid, has been abundantly reported to take part in inflammatory diseases. Recently, reports indicate that CRF family peptides may play an important role in the regulation of COX-2 under inflammatory conditions. Moreover, CRF receptors are involved in this process. This review aims to highlight the current novel findings on regulation of COX-2 by CRF family peptides in inflammation. Furthermore, the relevant mechanisms are discussed. PMID- 19292979 TI - DNA methyltransferase I is a mediator of doxorubicin-induced genotoxicity in human cancer cells. AB - Doxorubicin can induce the formation of extra-nuclear bodies during mitosis termed micronuclei but the underlying causes remain unknown. Here, we show that sub-lethal exposure to doxorubicin-induced micronuclei formation in human cancer cells but not in non-tumorigenic cells. Occurrence of micronuclei coincided with stability of DNMT1 upon doxorubicin assault, and DNMT1 was localized to the micronuclei structures. Furthermore, 5-aza-2'-deoxycytidine-mediated DNMT1 depletion or siDNMT1 knock-down attenuated the frequency of doxorubicin-induced micronucleated cells. Human DNMT1(-/-) cells displayed significantly fewer micronuclei compared to DNMT1(+/+) cells when challenged with doxorubicin, providing additional evidence for the involvement of DNMT1 in mediating such chromosomal aberrations. Collectively, our results demonstrate a role for DNMT1 in promoting DNA damage-induced genotoxicity in human cancer cells. DNMT1, recently identified as a candidate for doxorubicin-mediated cytotoxicity, is over expressed in various cancer cell types. We propose that DNMT1 levels in tumor cells may determine the effectiveness of doxorubicin in chemotherapy. PMID- 19292980 TI - Telmisartan, an angiotensin II type 1 receptor antagonist, attenuates T-type Ca2+ channel expression in neonatal rat cardiomyocytes. AB - Recently, it has been revealed that angiotensin II type 1 receptor (AT(1)) antagonists act as antiarrhythmic agents and that the T-type Ca2+ channel plays an important role in arrhythmia. However, it remains unclear how the T-type Ca2+ channel expression system is involved in angiotensin II-mediated arrhythmogenesis in cardiomyocytes. In this study, we investigated the effect of telmisartan, an AT(1) receptor antagonist, on transcriptional regulation of T-type Ca2+ channel isoform (Ca(v)3.1 and Ca(v)3.2) expression and cardiac contractility using rat neonatal cardiomyocytes. Cultured cardiomyocytes were stimulated with telmisartan and/or angiotensin II for 24 h. T-type Ca2+ currents (I(Ca.T)) were then measured with the patch clamp technique, while Ca(v)3.1 and Ca(v)3.2 mRNA expression were assessed by real-time PCR. Expression of Ca(v)3.1 and Ca(v)3.2 mRNA as well as I(Ca.T) current density in cardiomyocytes increased significantly after long-term application of angiotensin II (24 h), which was accompanied by extracellular signal-regulated kinase (ERK)1/2 and p38 mitogen-activated protein kinase (MAPK) phosphorylation. In contrast, telmisartan decreased Ca(v)3.1 and Ca(v)3.2 mRNA expression as well as I(Ca.T) in a dose-dependent manner in the absence of angiotensin II. In addition, the basal phosphorylation level of p38MAPK but not ERK1/2 was decreased by telmisartan in the absence of angiotensin II. Valsartan, an AT(1) receptor antagonist, did not mimic the action of telmisartan, while the action of telmisartan was completely blocked by valsartan. These results indicate that telmisartan attenuates T-type Ca2+ channel expression likely through p38MAPK activity in an agonist-independent manner, which suggests a novel pharmacological action of telmisartan. PMID- 19292981 TI - Preclinical assessment of proconvulsant drug activity and its relevance for predicting adverse events in humans. AB - Safety pharmacology studies, which are performed before first studies with investigational drugs in humans, often include experiments on proconvulsant drug activity, because such drugs are thought to promote seizures by decreasing seizure threshold. A commonly used model for the assessment of proconvulsant activity of investigational or marketed drugs is the timed intravenous pentylenetetrazole (PTZ) infusion seizure test, in which the latency to myoclonic or clonic seizures is determined by PTZ infusion in mice or rats. This test provides an extremely sensitive parametric method for assessing seizure threshold and allows detecting both anticonvulsant and proconvulsant drug effects. The aim of this review is to critically review the concept of "proconvulsant" drug activity and discuss data obtained by the PTZ and other seizure threshold tests as well as the various factors that may affect seizure threshold determinations. Furthermore, preclinical and clinical data on proconvulsant drug activity are compared. It is concluded that a battery of different tests is needed to provide the most reliable conclusions about the proconvulsant profile, if any, of drugs. Furthermore, misconceptions regarding proconvulsant drug effects, which can result in the undertreatment of brain diseases, are discussed. PMID- 19292982 TI - Immunohistochemical study on GABAergic system in salivary glands. AB - Gamma-aminobutyric acid (GABA) and its receptors are found in the central nervous system and several peripheral tissues. The purpose of this study was to determine the expression and distribution of GABA and glutamate decarboxylase (GAD), a GABA biosynthetic enzyme, in rat salivary gland. Western blot and real time quantitative RT-PCR revealed that GAD67 was the major isoform of GAD in the salivary glands. Furthermore, both GABA and GAD were detected around the acinar cells in the submandibular glands by immunohistochemical analysis. When both sympathetic and parasympathetic nerves related to the submandibular glands were denervated, the immunoreactivities of GABA and GAD were dramatically depressed, and levels of GAD67 and GABA significantly decreased. However, no morphological changes in the glands were observed after denervation. These results indicate that GAD67 is present around acinar cells in the salivary glands, and suggest that the GABAergic system in the glands is closely related to the autonomic nervous system. PMID- 19292983 TI - Unrepeatable extracellular Ca2+-dependent contractile effects of cyclopiazonic acid in rat vascular smooth muscle. AB - Cyclopiazonic acid (CPA), a specific reversible inhibitor of Ca(2+)-pumps in sarcoplasmic reticulum, causes a slowly developing and subsequently diminishing characteristic contraction in endothelium-denuded rat vascular smooth muscle. We recently found that CPA-induced contractions were not completely repeatable in endothelium-denuded rat aorta and superior mesenteric artery. 10 microM CPA induced contractions expressed as a percentage of 80 mM KCl-induced contraction were significantly decreased from 51.4+/-5.7% to 11.8+/-2.6% (P<0.0001) upon the second application in endothelium-denuded rat aorta, and this was not due to any irreversible cytotoxic effects of CPA. The decrease of CPA-induced contractile responses upon the second application was dependent on both types of blood vessels and doses of CPA upon the first application. CPA upon the second application in Ca(2+)-containing solutions did induce its characteristic contractions in the rings pretreated with Ca(2+)-free solutions or Ca(2+) entry blockers before and during its first application, suggesting that capacitative mode of Ca(2+) influx during the application of CPA might be responsible for the diminishment of contractions upon the second application. These data suggest that CPA by inducing a transient rise in cytosolic Ca(2+) level might cause a long lasting upregulation of Ca(2+) extrusion across the plasma membrane in vascular smooth muscle cells and thus accelerate Ca(2+) efflux over a prolonged period, leading to unrepeatable contractile effects of CPA. Such long-lasting upregulation of Ca(2+) extrusion may contribute to the regulation of excitability of vascular smooth muscle cells and protect the cells against excitotoxic injury. PMID- 19292984 TI - Zocor Forte (simvastatin) has a neuroprotective effect against LPS striatal dopaminergic terminals injury, whereas against MPP+ does not. AB - Due to their potential role in preventing further deterioration of Parkinson's disease, anti-inflammatory strategies have attracted great interest. In this context, some studies point out the possible protective effect of anti inflammatory compounds against the in vivo degeneration of dopaminergic neurons produced by lipopolysaccharide (LPS)-induced inflammatory processes and others. We have investigated the effect of the treatment of Zocor Forte (simvastatin) in LPS and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurodegenerative models to identify neuroprotective drugs for Parkinson's disease. We have perfused different concentrations of LPS or 1 mM 1-methyl-4-phenylpyridinium ion (MPP+) in the rat's striatum, 24 h after implanting a brain microdialysis probe, both with and without Zocor Forte (simvastatin) treatment. Results show that LPS perfusion produced a decrease in the basal release of dopamine. Forty-eight hours after implanting the probe, we have perfused 1 mM MPP+ to check the integrity of the dopaminergic terminals present around the cannula. Our model to study toxicity in the striatal dopaminergic terminals suggests that Zocor Forte (simvastatin) could prevent the neurotoxic damage produced by LPS, but not that produced by MPP+. PMID- 19292985 TI - Defining dose-response relationships in the therapeutic blockade of B7RP-1 dependent immune responses. AB - The ICOS (Inducible T cell Co-Stimulator)/B7RP-1 (B7-related protein 1) interaction is critical for the proper activation of a T lymphocyte. In this manuscript we describe a systematic in vivo approach to determine the level of blockade required to impair the generation of a T cell-dependent antibody response. We have developed an overall strategy for correlating drug exposure, target saturation, and efficacy in a biological response that can be generalized for most protein therapeutics. Using this strategy, we determined that low levels of B7RP-1 blockade are still sufficient to inhibit the immune response. These data suggest that contact between the T cell and the antigen-presenting cell during antigen presentation is much more sensitive to inhibition than previously believed and that ICOS/B7RP-1 blockade may be efficacious in the treatment of autoimmune diseases. PMID- 19292986 TI - Plasmodium chabaudi: expression of active recombinant chabaupain-1 and localization studies in Anopheles sp. AB - Plasmodium cysteine proteases have been shown to be immunogenic and are being used as malaria potential serodiagnostic markers and vaccine targets. Genes encoding two Plasmodium chabaudi cysteine proteases chabaupain-1 (CP-1) and chabaupain-2 (CP-2) were identified and further expressed in Escherichia coli. Solubilisation of recombinant CP-1 and CP-2 was achieved by decreasing the temperature of induction. Anopheles gambiae tissues infected with Plasmodium were analyzed by Western blotting using the anti-CP-1 antibody showing that CP-1 is only present in the A. gambiae midguts being absent from other infected mosquito biological material. Anti-CP-1 anti-serum recognized a 30 kDa band in P. chabaudi, Plasmodium berghei and Plasmodium yoelii lysates but does not recognize the recombinant CP-2 extracts suggesting high antibody specificity. PMID- 19292987 TI - Synthetic retinoid Am80 inhibits interaction of KLF5 with RAR alpha through inducing KLF5 dephosphorylation mediated by the PI3K/Akt signaling in vascular smooth muscle cells. AB - Kruppel-like factor 5 (KLF5) is known to physically interact with retinoic acid receptor-alpha (RAR alpha). Here, we show that Am80 inhibited the interaction between KLF5 and RAR alpha and this inhibitory effect was accompanied by the dephosphorylation of KLF5 in VSMCs. Treating VSMCs with LY294002, the PI3K/Akt inhibitor, abrogated Am80-induced KLF5 dephosphorylation and reversed Am80 induced suppression of interaction between KLF5 and RAR alpha, whereas treating vascular smooth muscle cells (VSMCs) with SB203580, the p38 kinase inhibitor, attenuated the interaction between KLF5 and RAR alpha. Constitutively active p38 kinase MKK6b infection prevented the KLF5 dephosphorylation induced by Am80. In conclusion, Am80 induces KLF5 dephosphorylation by activating PI3K/Akt signaling, and inhibits KLF5 phosphorylation by blocking p38 signaling, subsequently leading to the suppression of interaction of KLF5 with RAR alpha. PMID- 19292988 TI - Nicotine: a pro-angiogenic factor. AB - Angiogenesis, the formation of new blood vessels from pre-existing ones, is regulated by the balance between angiogenic activating and inhibiting factors. Recent evidence indicates that nicotine, an alkaloid compound, presents pro angiogenic effects in certain concentrations. Nicotine-induced angiogenesis results from the stimulation of non-neuronal nicotinic acetylcholinergic receptors (particularly alpha 7-NAChR subtype) and involves growth factor secretion and activation of intracellular signalling pathways. Although nicotine is a constituent of tobacco smoke, its contribution towards pathophysiology of several tobacco-associated pathologies is controversial. Nowadays, nicotine is used in tobacco smoke cessation programs, but, again, its use is still a matter of discordance regarding cardiovascular morbidity. Nevertheless, given the established effects of nicotine-induced angiogenesis, this alkaloid might be of therapeutic value in situations of inefficient angiogenesis, such as peripheral vascular impairment and tissue ischemia. Conversely, blockade of nicotine pathway can be helpful in complications exhibiting excessive angiogenesis, namely in diabetic retinopathy and nephropathy or neoplasia. Knowing that nicotine is involved in many angiogenic-associated disorders with a high prevalence in the western world, and that this alkaloid widely used in smoke cessation programs, elucidation of the action of nicotine on the vascularization process is of utmost importance. The current paper provides an overview of the effects of nicotine in angiogenesis. PMID- 19292989 TI - Xeomin is free from complexing proteins. AB - In contrast to the other botulinum toxin products Xeomin only contains the 150kD neurotoxin without complexing proteins which have no therapeutic function and don't influence the diffusion of the neurotoxin. In large clinical Phase III studies (blepharospasm and cervical dystonia) Xeomin showed the same efficacy and profile of adverse events as Botox. Whereas competing product must be stored refrigerated, Xeomin is stable for 3years at room temperature. PMID- 19292991 TI - The role of neuro-immune cross-talk in the regulation of inflammation and remodelling in asthma. AB - Despite recent advances in the development of anti-asthmatic medication, asthma continues to be a major health problem worldwide. The symptoms of asthmatic patients include wheezing, chest tightness, cough and shortness of breath, which, together with airway hyperresponiveness, previously have been attributed to a dysfunction of airway nerves. However, research in the last two decades identified Th2-sensitization and the subsequent allergic reaction to innocuous environmental antigens as a basic immunological mechanism leading to chronic airway inflammation. Recent evidence suggests that the development of allergic asthma is influenced by events and circumstances in early childhood and even in utero. Allergen, ozone or stress exposure, as well as RSV infection in early life could be able to induce irreversible changes in the developing epithelial mesenchymal trophic unit of the airways. The co-existence of chronic inflammation and neural dysfunction have recently drawn attention to the involvement of interaction pathways between the nervous and the immune system in the airways. Intensive basic research has accumulated morphological as well as functional evidence for the interaction between nerves and immune cells. Neuropeptides and neurotrophins have come into focus of attention as the key mediators of neuro immune interactions, which lead to the development of several pharmacological compounds specifically targeting these molecules. This review will integrate our current knowledge on the involvement of neuro-immune pathways in asthma on the cellular and molecular level. It will summarize the results of pharmacological studies addressing the potential of neuropeptides and neurotrophins as novel therapeutic targets in asthma. PMID- 19292992 TI - Self-exploding microcapsules for pulsed drug delivery. PMID- 19292993 TI - Pharmacogenomics in non-small-cell lung cancer chemotherapy. AB - The disappointing results in long-term survival of patients who have a resectable non-small cell lung cancer (NSCLC) may reflect the lack of knowledge on the way by which molecular abnormalities in neoplastic cells affect responsiveness to adjuvant therapy. This issue deserves intensive investigation to select methodological approaches for a new generation of chemotherapeutic strategies. Remarkable advances in the understanding of NSCLC biology have been made, including the discovery of critical mutations in oncogenes (i.e. K-Ras and c myc), as well as the loss of tumor-suppressor genes, such as TP53, p16(INK4) or Rb. Other studies demonstrated the role of mutations or deregulation of the expression of several molecular determinants involved in cell cycle control such as epidermal growth factor receptor (EGFR). All these characteristics, as well as alterations in gene products directly related to drug activity, might contribute to the aggressive behaviour of NSCLC. The future challenge of chemotherapy of NSCLC relies on the identification of molecular markers that are predictive of drug sensitivity and are helpful in the selection of chemotherapeutic agents best suited to the individual patient. Other intriguing issues will be the identification of the optimal drug sequence in combination regimens and the pharmacogenetics of severe toxicities. Moreover, due to the developments of novel technologies to decipher genetic alterations involved in tumor progression, new agents are gaining momentum, including inhibitors of intracellular signal transduction, and a large body of research, using prospective clinical trials, should be devoted to this area. PMID- 19292994 TI - Characterization of biominerals in the radula teeth of the chiton, Acanthopleura hirtosa. AB - Understanding biomineralization processes provides a route to the formation of novel biomimetic materials with potential applications in fields from medicine to materials engineering. The teeth of chitons (marine molluscs) represent an excellent example of a composite biomineralized structure, comprising variable layers of iron oxide, iron oxyhydroxide and apatite. Previous studies of fully mineralized teeth using X-ray diffraction, Raman spectroscopy and scanning electron microscopy (SEM) have hinted at the underlying microstructure, but have lacked the resolution to provide vital information on fine scale structure, particularly at interfaces. While transmission electron microscopy (TEM) is capable of providing this information, difficulties in producing suitable samples from the hard, complex biocomposite have hindered progress. To overcome this problem we have used focused ion beam (FIB) processing to prepare precisely oriented sections across interfaces in fully mineralized teeth. In particular, the composite structure is found to be more complex than previously reported, with additional phases (goethite and amorphous apatite) and interface detail observed. This combination of FIB processing and TEM analysis has enabled us to investigate the structural and compositional properties of this complex biocomposite at higher resolution than previously reported and has the potential to significantly enhance future studies of biomineralization in these animals. PMID- 19292995 TI - Polyploid origins in Gynostemma pentaphyllum (Cucurbitaceae) inferred from multiple gene sequences. AB - The genus Gynostemma (Cucurbitaceae) constitutes a polyploid group of perennial creeping herbs, in whose evolution polyploidization is a key component. With the largest variety of cytotypes (2n=22, 44, 66 and 88) in Gynostemma, G. pentaphyllum is also the most widespread species in this genus. In the present study, we inferred the origins of polyploids in G. pentaphyllum using sequences of the plastid intergenic spacers (trnL-trnF, psbB-psbF and rpl20-rps12) and cloned DNA sequences from two nuclear regions (RPB2 and nrDNA ITS). Phylogenetic analyses of the separate and the combined nuclear gene datasets all supported autoploid origins of polyploids in G. pentaphyllum. Three polyploid populations were more closely related, indicating that significant genetic differentiation may have occurred between diploids and polyploids. We concluded that polyploidization might be an important evolutionary mechanism in the diversification of G. pentaphyllum. On the other hand, no chloroplast DNA (cpDNA) variation was detected in ingroups except the octoploid DL 8x, which possessed a different cpDNA haplotype from the other populations of G. pentaphyllum. This can be explained by limited sample sizes, possible extinction of its diploid progenitors and/or the occurrence of chloroplast transfer through hybridization with other Gynostemma species. However, the distribution of cytotypes in G. pentaphyllum was not as typical as many other autopolyploid complexes. Polyploidization failed to contribute significantly to the expansion of its geographic range. The geographic distribution of diploids and polyploids in G. pentaphyllum may be associated with the past ecological environments of different areas, especially during the glacial period. PMID- 19292996 TI - Maternal care determines rapid effects of stress mediators on synaptic plasticity in adult rat hippocampal dentate gyrus. AB - Maternal care in the rat influences hippocampal development, synaptic plasticity and cognition. Previous studies, however, have examined animals under minimally stressful conditions. Here we tested the hypothesis that maternal care influences hippocampal function differently when this structure is exposed to corticosteroid and noradrenergic hormones, which are elevated during the early phase of a stress response. In the adult male offspring of Long-Evans dams characterised as high or low in maternal care (high LG and low LG) we (1) examined basal dendritic morphology in the dentate gyrus by Golgi staining; (2) investigated rapid modulation of in vitro long term-potentiation (LTP) in the dentate gyrus by glucocorticoid and beta-adrenergic stimulation; (3) examined hippocampal and amygdala-dependent learning under stress using contextual and cued fear conditioning. We found differences in hippocampal dentate gyrus morphology in adult offspring of high and low LG mothers, with less dendritic complexity in low LG offspring. Under basal conditions LTP was lower in slices from low compared with high LG offspring. Hippocampal LTP was rapidly increased by either corticosterone (100 nM) or isoproterenol (1.0 microM) in low LG offspring, suggesting improved dentate plasticity during stress. This was mirrored in hippocampal but not amygdala-dependent learning, as low LG offspring showed enhanced contextual but not cued fear conditioning. We suggest that decreased pup LG during postnatal life may be adaptive in high-threat environments, potentially enhancing hippocampal function in the offspring under conditions of adversity. PMID- 19292997 TI - Grey-matter differences related to true and false recognition of emotionally charged stimuli - a voxel based morphometry study. AB - The issue concerning the neuronal basis of true and false recognition is still a subject of extensive debate. In the present study voxel based morphometry (VBM) was used to examine structural brain correlates of these processes. Since several studies indicate that emotional content facilitates false recognition we decided to use emotional stimuli. Behavioral measures, i.e., true and false recognition rates were used as covariants in VBM analyses. VBM results indicated that the true recognition correlated positively with grey-matter (GM) density in bilateral amygdala, anterior cingulate and middle temporal gyrus, i.e., brain regions, involved in the memory of emotional material, as revealed by fMRI results. False recognition correlated negatively with GM density in prefrontal areas (BA47 and BA9), supporting the role of the prefrontal cortex in monitoring retrieval and limiting the probability of false recognition. Thus our VBM findings (i) point to the brain structures critical for correct and false emotional memory and (ii) disclose structural differences between the neural bases of these two types of memory. PMID- 19292998 TI - Characterization of two putative fibronectin-binding proteins of Clostridium perfringens. AB - Clostridium perfringens is a Gram-positive anaerobic pathogen that causes gas gangrene and food poisoning in humans and animals. Genomic analysis of C. perfringens strain 13 revealed that this bacterium contains two genes (CPE0737 and CPE1847) that encode putative fibronectin (Fn)-binding proteins (Fbps). These genes, named fbpA and fbpB, were found to be constitutively expressed in all three strains (13, NCTC8237, CPN50) of C. perfringens, isolated from gas gangrene of human, that were tested. Both fbpA and fbpB were cloned and His-tagged, recombinant FbpA (rFbpA) and recombinant FbpB (rFbpB) were purified by Ni(2+) Sepharose column chromatography from transformed Escherichia coli. These recombinant Fbps were shown to bind to Fn, purified from human serum, in a ligand blotting assay. Additionally, Fn bound to these rFbps in a dose-dependent manner in an enzyme-linked immunosorbent assay. Furthermore, it was found that pre incubation of Fn with either rFbpA or rFbpB inhibited the binding of Fn to C. perfringens cells. PMID- 19292999 TI - Antimicrobial susceptibility of anaerobic bacteria in Bulgaria. AB - OBJECTIVES: The antimicrobial susceptibility of anaerobic bacteria isolated from clinical specimens in the referent for Bulgaria anaerobic laboratory was studied in a period of 25 years (1983-2007). METHODS: NCCLS-recommended agar dilution methods were used. beta-lactamase activity was determined with nitrocefin discs. RESULTS: The 29 antimicrobial agents included in the study were divided according to their in vitro activity against the anaerobic isolates into 4 main groups for guiding empirical treatment: 1st group of metronidazole, chloramphenicol, meropenem, imipenem and combinations of beta-lactam antibiotics with sulbactam- with high activity and drugs of choice for treatment; 2nd group--clindamycin, cefoxitin, carbenicillin/and azlocillin, piperacillin/--with a good activity and low percent of resistant strains; 3rd group--of tetracycline and erythromycin with higher percent of resistant strains including the new macrolides as josamycin, clarithromycin, roxithromycin and azithromycin; 4th group- penicillins/ampicillin, amoxicillin, penicillin/and cephalosporins/cefamandole, cefazolin, cefotaxime and cefoperazone/--not suitable for treatment of infections including Bacteroides fragilis group strains, with a very high percent of resistant strains, probably due to beta-lactamase activity in most of the strains. CONCLUSION: A continued updating and a follow-up in the changes of antibiotic susceptibility are necessary in every country as resistance patterns vary not only between geographical regions but also even among medical centers and hospitals which may be connected with differences in antibiotic usage in man and animals. PMID- 19293000 TI - Intestinally implanted Nippostrongylus brasiliensis adult worms decrease serum paraoxonase-1 activity in rats. AB - We have previously reported a significant decrease in serum PON1 activity after Nippostrongylus brasiliensis infection in Wistar rats in association with the inflammatory response mounted against the parasite in the migratory phase of infection. However, the roles of intestinal phase and the associated oxidative stress during N. brasiliensis infection on PON1 activity have not yet been elucidated. In the present study, we observed a significant reduction in serum paraoxonase and arylesterase activity on days 6 and 9 post-implantation with N. brasiliensis adult worms in the absence of a significant increase in various serum pro-inflammatory cytokines. In addition, provision of the antioxidant butylated hydroxyanisole (BHA) to adult worm-implanted rats did not ameliorate the reduction in PON1 activity. Due to the prolonged intestinal phase of gastrointestinal nematode infections, alterations in PON1 activity during this phase need to be further examined to elucidate the mechanism of alteration in PON1 activity. PMID- 19293001 TI - Morphology of Luolishania longicruris (Lower Cambrian, Chengjiang Lagerstatte, SW China) and the phylogenetic relationships within lobopodians. AB - New material of the lobopodian Luolishania longicruris has been recovered from the Lower Cambrian Chengjiang Lagerstatte, southwest China. The specimens throw new light on several morphological features of the species, including the paired antenniform outgrowths, eyes, head shield, setae and other cuticular projections, as well as the differentiated sclerites, appendages, claws, and lobopod interspaces. L. longicruris shows well developed tagmosis: a distinct head and a trunk divided into two sections. The new data allow a revised comparison with other lobopodians. Miraluolishania haikouensis Liu et al., 2004 is considered to be a junior synonym of L. longicruris Hou and Chen, 1989. Evidence from gut filling and specialized morphological characters indicates that L. longicruris may have had a filter feeding lifestyle. A new cladistic analysis suggests that fossil lobopodians are paraphyletic or even polyphyletic and L. longicruris may be an important representative of the stem lineage leading to arthropods. PMID- 19293002 TI - A cell-based screening assay for Natural Killer cell activity. AB - Natural Killer (NK) cells are important in the first response against viruses and tumours. Compounds that modulate human NK cell activity offer interesting prophylactic and therapeutic options, however, a systematic screening tool is lacking. Development of suitable NK cell lines or receptor-based assays is hindered by the highly complicated regulation of the different NK cell subsets by multiple receptors. Here, we describe a cell-based flowcytometric activity assay adapted to identify NK cell modulating compounds. Fresh human peripheral blood mononuclear cells (PBMC) were incubated with NK-sensitive K562 target cells labelled with 5-(6)-carboxyfluorescein succinimidyl ester, followed by DNA labelling with propidium iodide to identify dead cells. The assay demonstrated a good performance with an average Z'-factor of 0.6 and over 95% of the assays fulfilled the quality criteria, suggesting that it is possible to use a complex system with two different cell types to screen compounds. A large number of (natural) compounds and extracts were tested and normalized to the positive control, Interleukin-2. Promising and less promising compounds were distinguished. Effectiveness of compounds was based on the augmentation of NK cell activity as well as the number of responding subjects. To conclude the assay is robust, reliable and can be used for functional screening of natural compounds modulating NK cell activity. PMID- 19293003 TI - Valnemulin downregulates nitric oxide, prostaglandin E2, and cytokine production via inhibition of NF-kappaB and MAPK activity. AB - Valnemulin is a pleuromutilin antibiotic used in clinics for the treatment of various infections. We studied the in vitro anti-inflammatory effects of valnemulin and associated signal transduction mechanisms in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. We found that valnemulin inhibited nitric oxide (NO), prostaglandin E2 (PGE2), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and increased interleukin-10 (IL-10) production. Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression were also inhibited by valnemulin. We further observed that valnemulin prevented the LPS-induced NF-kappaB translocation from the cytoplasm into the nucleus. Valnemulin also blocked phosphorylation of three mitogen-activated protein kinases (MAPKs): extracellular signal receptor-activated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK). Our data indicate that valnemulin may have therapeutic anti-inflammatory effects independent of its antibacterial activity. PMID- 19293004 TI - Autoimmune neutropenia in adults. AB - Autoimmune neutropenias (AIN) in adults are a heterogeneous group of diseases with clinical manifestations varying from being asymptomatic to having infectious complications with considerable morbidity and mortality. They are characterized by autoantibodies directed against neutrophils, resulting in destruction of neutrophils. AIN can be divided into two forms. In primary AIN, neutropenia is usually the sole hematologic abnormality and it is more common in children. Secondary AIN, which is more prevalent in adults, is associated with underlying autoimmune diseases, malignancies, infections, particularly viral, neurological diseases or drug exposure. This article is an overview of these conditions with emphasis on secondary AIN; it also discusses the available serological methods for antibody detection and recent therapeutic developments including colony stimulating factors, rituximab and Campath-1H. PMID- 19293005 TI - Tests of gastric neuromuscular function. AB - Tests of gastric neuromuscular function are used to evaluate patients with symptoms referable to the upper digestive tract. These symptoms can be associated with alterations in the rates of gastric emptying, impaired accommodation, heightened gastric sensation, or alterations in gastric myoelectrical function and contractility. Management of gastric neuromuscular disorders requires an understanding of pathophysiology and treatment options as well as the appropriate use and interpretation of diagnostic tests. These tests include measures of gastric emptying; contractility; electrical activity; regional gastric motility of the fundus, antrum, and pylorus; and tests of sensation and compliance. Tests are also being developed to improve our understanding of the afferent sensory pathways from the stomach to the central nervous system that mediate gastric sensation in health and gastric disorders. This article reviews tests of gastric function and provides a basic description of the tests, the methodologies behind them, descriptions of the physiology that they assess, and their clinical utility. PMID- 19293006 TI - Hidradenitis suppurativa: a comprehensive review. AB - Hidradenitis suppurativa, also known as acne inversa, is a chronic, often debilitating disease primarily affecting the axillae, perineum, and inframammary regions. Prevalence rates of up to 4% have been estimated. Our understanding of the disease has changed over time. It is now considered a disease of follicular occlusion rather than an inflammatory or infectious process of the apocrine glands. Clinically, the disease often presents with tender subcutaneous nodules beginning around puberty. The nodules may spontaneously rupture or coalesce, forming painful, deep dermal abscesses. Eventually, fibrosis and the formation of extensive sinus tracts may result. The location of the lesions may lead to social embarrassment and the failure to seek medical treatment. Therapies in the past have consisted of long-term antibiotics, antiandrogens, and surgery. New treatments like tumor necrosis factor-alfa inhibitors have given clinicians more options against this difficult disease. PMID- 19293007 TI - Hair grooming practices and central centrifugal cicatricial alopecia. AB - BACKGROUND: The cause of central centrifugal cicatricial alopecia (CCCA) in African American women remains to be elucidated. OBJECTIVE: This study was designed to determine the hair-grooming practices in African American women with and without CCCA and to evaluate possible etiologic factors. METHODS: Utilizing a novel survey instrument, the Hair Grooming Assessment Survey, we performed a retrospective comparative survey of the hair-grooming practices of two populations of African American women seen and evaluated at the Department of Dermatology, Henry Ford Hospital in Detroit, MI, between 2000 and 2007. The case group were women with clinical and histologic diagnosis of CCCA, and the control group were those without a history of alopecia. RESULTS: All 101 surveys that were returned were analyzed (51 from the case group and 50 from the control group). A strong association was found between the use of both sewn-in hair weaving and cornrow or braided hairstyles with artificial hair extensions and CCCA (P < .04, P < .03, respectively). Similarly, women with CCCA were more likely to report a history of "damage", typically defined as uncomfortable pulling and tenderness, from both sewn-in and glued-in weaves, and from cornrow or braided hairstyles with artificial hair extensions (P < .001, P < .02, and P < .03, respectively). In contrast to previous anecdotal beliefs, no correlation was found between the use of either hot combing or hair relaxers and the development of CCCA. LIMITATIONS: Results are limited by patient recall of past hair grooming practices. Also, as hair grooming practices may vary by geographic region, these results may not be generalized to all women of African descent. CONCLUSION: There is a clear difference in both quantitative and qualitative hair grooming practices among African American women with CCCA. PMID- 19293009 TI - Phenol peels as a novel therapeutic approach for actinic keratosis and Bowen disease: prospective pilot trial with assessment of clinical, histologic, and immunohistochemical correlations. AB - BACKGROUND: Although chemical peels may be used for precancerous lesions, no histologic or immunohistochemical studies have been performed to validate clinical impressions and/or outcome. OBJECTIVE: Our purpose was to investigate the efficacy and prognostic relevance of phenol peels in Japanese patients with actinic keratosis and Bowen disease using clinical and histologic criteria. METHODS: A total of 46 patients were treated with phenol peels, and followed up for at least 1 year after treatment. Biopsy specimens were taken before and after treatment. Cases of complete response were classified by the number of treatment sessions. We evaluated parameters for epidermal thickness, proliferation, dysplasia, and apoptosis, and clinical characteristics to correlate phenol peels with assessments of efficacy, patient-selection criteria, and risk for transformation to cutaneous squamous cell carcinoma. RESULTS: There were 39 (84.8%) patients with a complete response after one to 8 treatment sessions. Statistically, differences in clinical improvement with peels and the number of treatment sessions correlated with histology, personal history of skin cancer, tumor thickness, and cyclin A expression. LIMITATIONS: This study was a prospective pilot trial. Blinded, placebo-controlled, randomized studies would be ideal. CONCLUSION: We conclude that phenol peels are very effective for treating precancerous lesions of actinic keratosis and Bowen disease. In addition, our study clearly demonstrates that tumor thickness and cyclin A could be specific and useful markers as adjunctive diagnostic tools to predict the efficacy of phenol treatment of these lesions. PMID- 19293008 TI - A randomized double-blind trial of intravenous immunoglobulin for pemphigus. AB - BACKGROUND: Pemphigus is a rare life-threatening intractable autoimmune blistering disease caused by IgG autoantibodies to desmogleins. It has been difficult to conduct a double-blind clinical study for pemphigus partly because, in a placebo group, appropriate treatment often must be provided when the disease flares. OBJECTIVE: A multicenter, randomized, placebo-controlled, double-blind trial was conducted to investigate the therapeutic effect of a single cycle of high-dose intravenous immunoglobulin (400, 200, or 0 mg/kg/d) administered over 5 consecutive days in patients relatively resistant to systemic steroids. METHODS: We evaluated efficacy with time to escape from the protocol as a novel primary end point, and pemphigus activity score, antidesmoglein enzyme-linked immunosorbent assay scores, and safety as secondary end points. RESULTS: We enrolled 61 patients with pemphigus vulgaris or pemphigus foliaceus who did not respond to prednisolone (> or =20 mg/d). Time to escape from the protocol was significantly prolonged in the 400-mg group compared with the placebo group (P < .001), and a dose-response relationship among the 3 treatment groups was observed (P < .001). Disease activity and enzyme-linked immunosorbent assay scores were significantly lower in the 400-mg group than in the other groups (P < .05 on day 43, P < .01 on day 85). There was no significant difference in the safety end point among the 3 treatment groups. LIMITATION: Prednisolone at 20 mg/d or more may not be high enough to define steroid resistance. CONCLUSION: Intravenous immunoglobulin (400 mg/kg/d for 5 d) in a single cycle is an effective and safe treatment for patients with pemphigus who are relatively resistant to systemic steroids. Time to escape from the protocol is a useful indicator for evaluation in randomized, placebo-controlled, double-blind studies of rare and serious diseases. PMID- 19293010 TI - Efficacy and safety of pulsed dye laser treatment for cutaneous discoid lupus erythematosus. AB - BACKGROUND: Treatment of chronic discoid lupus erythematosus (CDLE) with a pulsed dye laser (PDL) has shown promising results, although outcomes in previous studies were not validated and laser parameters were inconsistent. OBJECTIVE: We conducted an open prospective study to assess the efficacy and safety of PDL for the treatment of recalcitrant CDLE, using a validated scoring method and a fixed treatment schedule. METHODS: Twelve patients with active CDLE lesions were treated with PDL (585 nm, fluence 5.5 J/cm(2), spot size 7 mm) 3 times with an interval of 6 weeks followed by a 6-week follow-up period. Treatment outcomes were evaluated by 3 observers using the validated Cutaneous Lupus Erythematosus Disease Area and Severity Index (CLASI). Cosmetic results and adverse events were recorded. RESULTS: A significant decline in "active" CLASI was observed after 6 weeks, after 12 weeks, and at follow-up. Baseline active CLASI was 4.4 +/- 0.2 (mean +/- SEM), reaching 1.3 +/- 0.3 after follow-up (P < .0001). Individual scores for erythema and scaling/hypertrophy significantly declined 6 weeks after treatment. The "damage" CLASI (dyspigmentation, scarring, and atrophy) did not show any significant change during or after therapy. The observed clinical improvement was confirmed by two independent observers by clinical assessment of photographs (r = 0.87 and r = 0.89; both P < .05). The treatment was well tolerated, only minimal pain was reported, and the cosmetic result was fair. LIMITATIONS: Small sample size and short follow-up duration were limitations. CONCLUSION: PDL treatment is an effective and safe therapy for patients with refractory CDLE. PMID- 19293011 TI - Classification of cutaneous intravascular breast cancer metastases based on immunolabeling for blood and lymph vessels. AB - BACKGROUND: Intravascular breast cancer metastases to the skin can have several clinically distinct manifestations. Carcinoma telangiectoides, which presents as an erythematous patch with prominent telangiectasias or lymphangioma circumscriptum-like lesions, is a rare manifestation of cutaneous metastatic breast cancer and has been proposed to spread via dermal blood vessels. Carcinoma erysipelatoides, which presents as an erysipelas-like patch or plaque, has been proposed to spread via lymphatics. Clinical variants with nodular lesions that show tumor cells within vessels and in the extravascular space are seen more commonly. It has not been demonstrated conclusively whether dermal blood vessels or whether dermal lymph vessels are principally involved in these clinically distinct forms of cutaneous breast cancer metastases. OBJECTIVES: We sought to determine if carcinoma telangiectoides affects predominantly dermal blood vessels and if carcinoma erysipelatoides affects predominantly dermal lymph vessels. METHODS: Serial sections of biopsy specimens from patients with a characteristic clinical presentation of carcinoma telangiectoides and carcinoma erysipelatoides were labeled for cytokeratin to identify malignant cells. Subsequently, these sections were labeled for CD31 (a marker for blood and lymph vessels) and podoplanin (a marker for lymph vessels, but not for blood vessels), to differentiate blood vessels from lymph vessels in these sections. RESULTS: Sections from carcinoma telangiectoides showed malignant tumor cells strictly within dermal blood vessels, but not in lymph vessels. In contrast, sections from carcinoma erysipelatoides showed malignant tumor cells strictly in dermal lymphatics. LIMITATIONS: We used typical clinical cases to demonstrate the distinct involvement of blood and lymph vessels in these variants of cutaneous metastatic breast cancer. A larger case series is needed to confirm these findings. CONCLUSIONS: Immunolabeling for CD31 and podoplanin of cutaneous lesions of metastatic breast cancer confirms the spread of tumor cells predominantly via lymphatics in carcinoma erysipelatoides and predominantly via blood vessels in carcinoma telangiectoides. PMID- 19293012 TI - Reflectance confocal microscopy for the in vivo detection of Treponema pallidum in skin lesions of secondary syphilis. AB - BACKGROUND: Secondary syphilis is a diagnostic challenge that relies on microscopic and laboratory tests. OBJECTIVE: We sought to assess the usefulness of in vivo reflectance confocal microscopy (RCM) to detect Treponema pallidum in lesions suggestive of secondary syphilis. METHODS: Macular and papular skin lesions from 3 patients clinically suggestive of secondary syphilis were imaged by RCM and confirmed by skin punch biopsy. RESULTS: In all lesions RCM demonstrated elongated small bright particles with a spiral shape intermingled with the keratinocytes. These features corresponded with immunohistochemical findings that revealed several spirochetes infiltrating the epidermis. LIMITATIONS: Unlike immunohistochemistry, RCM did not visualize T pallidum in the dermis and vascular walls because of limited imaging depth. The specificity and sensitivity of this technique need to be assessed. CONCLUSION: RCM may be an effective diagnostic tool for in vivo real-time imaging of T pallidum in skin lesions of secondary syphilis, and seems to correlate well with immunohistochemistry. PMID- 19293013 TI - Central centrifugal cicatricial alopecia: past, present, and future. AB - Clinical scarring alopecia in African American women has been recognized for years. The classification of this unique form of alopecia dates back to Lopresti, who first described the entity called "hot comb alopecia." More recently, the term "central centrifugal cicatricial alopecia" has been adopted to describe a progressive vertex-centered alopecia most common in women of African descent. While this form of hair loss is widely recognized, and may even be on the rise, the causes of central centrifugal cicatricial alopecia are a constant source of debate and remain to be elucidated. This review outlines the descriptive evolution of central centrifugal cicatricial alopecia and the historical controversies ascribed to its pathoetiology; it also examines African hair structure and discusses how hair structure along with common physical and chemical implements utilized by individuals with African hair type may play a causal role in the development of central centrifugal cicatricial alopecia. PMID- 19293014 TI - Enzyme-linked immunosorbent assay and immunobullous disease. PMID- 19293016 TI - Trichodaganomania: the compulsive habit of biting one's own hair. AB - A 17-year-old male presented with patches of hair loss on both forearms with comorbid depression and anxiety disorders. He had irresistible urges to bite his hair on easily approachable sites and endorsed biting his hair during periods of severe anxiety and boredom, resulting in noticeable patches of hair loss. There was no evidence of trichotillomanic behavior or any other skin pathology resulting in loss of hair at those sites. Microscopy of the bitten end of hairs showed smooth, sharply demarcated, blunted shafts. Trichodaganomania (biting one's own hair) should not be confused with trichotillomania, trichotemnomania, or trichoteiromania. A psychodermatologic evaluation is warranted in the evaluation of these cases. PMID- 19293015 TI - Characteristics of swimming pools with high rates of objectively measured sunscreen use. AB - BACKGROUND: A combination of verbal reports, observations, and physical assessments can improve understanding of prevention programs. OBJECTIVE: Skin swabbing techniques were used to detect the presence of sunscreen as part of a sun protection measurement study at 16 swimming pools. Three pools demonstrated much higher sunscreen use (>95%) than the others (47.1%). This paper compares these three pools to the other 13 to examine whether they have common features and if different sources of data can help interpret the findings. METHODS: Data were collected from skin swabs detecting the presence of sunscreen; observations of participants; observations of pool environments; and surveys. RESULTS: Pool observations showed a higher use of shade structures, sun safety signs, and other supporting items at the three high-sunscreen use pools. These three pools had significantly more year-round and long-term employees than did the other 13 pools. LIMITATIONS: Sunscreen characteristics could not be determined using the swabbing technique. Publicity about the study, or the weather, may have influenced behaviors at the pools. CONCLUSIONS: Supportive environments were associated with consistent high levels of sunscreen use. This study also confirms the importance of using multiple data sources to interpret findings. PMID- 19293017 TI - Bullous papular-purpuric gloves and socks syndrome in a 42-year-old female: molecular detection of parvovirus B19 DNA in lesional skin. AB - Papular-purpuric gloves and socks syndrome is a self-limited febrile illness of children and young adults. Only 50 well documented cases have been reported, most of which were associated with parvovirus B19 infection. Molecular detection of the virus from lesional skin has been described in only 5 patients. The syndrome is characterized by a papular-purpuric edematous rash in a distinct "gloves and socks" distribution. Extracutaneous manifestations are usually mild and transient. We report a 42-year-old female with a highly unusual expression of the syndrome, including bullous lesions, lingual aphthae, and conjunctivitis, accompanied by arthritis and a high-grade fever. She had immunoglobulin M and immunoglobulin G antibodies to parvovirus B19, which was detected from lesional skin with the use of polymerase chain reaction for the first time in such a clinical constellation. Because parvovirus B19 infections are generally more severe in adults, we suspect that the unusual disease expression in our patient was related to being more than 40 years of age. PMID- 19293018 TI - Fixed solar urticaria with delayed onset. AB - Fixed solar urticaria (FSU) is an extremely rare type of solar urticaria characterized by urticarial wheals appearing frequently confined to fixed areas of the skin. After a few minutes of exposure to sunlight or other sources of radiation, urticarial lesions can usually be induced exclusively in the same localization. We report a case of delayed onset FSU occurring 6 hours after exposure to ultraviolet A and B light. PMID- 19293019 TI - A case of hypermelanosis in a phylloid pattern. AB - Pigmentary anomalies display a variety of different patterns. The phylloid pattern is characterized by a leaf-shaped arrangement reminiscent of floral ornaments. We describe a 20-year-old Korean man with hyperpigmented oblong patches and atypically short, thick hairs in a phylloid pattern on the face, the left shoulder, and the left side of the trunk. Associated musculoskeletal anomalies included equinovarus deformities, bilateral club foot, lumbar lordosis, spina bifida, and relative hypertrophy of the right thigh muscles. PMID- 19293020 TI - Penicillamine-induced elastosis of the mucosal lip. AB - Long-term penicillamine therapy has been associated with alterations in dermal elastic tissue. Well-described associated dermatoses include pseudo pseudoxanthoma elasticum, acquired cutis laxa, elastosis perforans serpiginosa, and anetoderma. Histologically, "lumpy-bumpy"" or "bramble-bush"" morphologic changes of elastic fibers in the dermis are characteristic. Previous reports of these findings in normal-appearing skin and internal organs suggest a systemic elastolytic process. Here we report an unusual case of penicillamine-induced elastosis affecting the mucosa of the lip with characteristic histologic features. PMID- 19293021 TI - Mortality of bullous pemphigoid. PMID- 19293022 TI - Metastatic basal cell carcinoma: case unproven. PMID- 19293023 TI - Lack of efficacy of topical latanoprost and bimatoprost ophthalmic solutions in promoting eyelash growth in patients with alopecia areata. PMID- 19293024 TI - Follow: the Raptiva (efalizumab) Pregnancy Registry. PMID- 19293025 TI - Evaluation of oral zinc sulfate effect on recalcitrant multiple viral warts: a randomized placebo-controlled clinical trial. PMID- 19293026 TI - Toxic epidermal necrolysis secondary to emergency contraceptive pills. PMID- 19293027 TI - Development of primary varicella infection during infliximab treatment for psoriasis. PMID- 19293028 TI - Generalized essential telangiectasia with predilection for surgical scar. PMID- 19293029 TI - Cutaneous cholesterol embolization. PMID- 19293030 TI - Systematized linear porokeratosis: a rare variant of diffuse porokeratosis with good response to systemic acitretin. PMID- 19293031 TI - Alfa-1-antitrypsin deficiency panniculitis acquired after liver transplant and successfully treated with retransplant. PMID- 19293032 TI - Legal immunity? Wrong diagnosis, wrong prescription: another lesson from enterovirus vaccines. PMID- 19293033 TI - Induction of Akt activity by chemotherapy confers acquired resistance. AB - Resistance to chemotherapy is a major cause of treatment failure in human cancer. Accumulating evidence has indicated that the acquisition of resistance to chemotherapeutic drugs involves the activation of the PI3K/Akt pathway. Modulating Akt activity in response to chemotherapy has been observed often in chemoresistant cancers. The potential molecular mechanisms by which chemotherapeutic agents activate the PI3K/Akt pathway are emerging. Activation of this pathway evades the cytotoxic effects of chemotherapeutic agents via regulation of essential cellular functions such as protein synthesis, antiapoptosis, survival and proliferation in cancer. How chemotherapeutic agents induce Akt activation and how activated Akt confers chemoresistance through regulation of signaling networks are discussed in this review. Combining PI3K/Akt inhibitors with standard chemotherapy has been successful in increasing the efficacy of chemotherapeutic agents both in vivo and in vitro. Several small molecules have been developed to specifically target PI3K/Akt and other components of this pathway, which in combination with chemotherapy may be a valid approach to overcome therapeutic resistance. We propose several feedback and feedforward regulatory mechanisms of signaling networks for maintenance of the Akt activity for cell survival. These regulatory mechanisms may limit the efficacy of PI3K/Akt-targeted therapy; therefore, disruption of these mechanisms may be an effective strategy for development of novel anti-cancer therapies. PMID- 19293034 TI - Abdominal tuberculosis in southeastern Taiwan: 20 years of experience. AB - BACKGROUND/PURPOSE: Abdominal tuberculosis (TB) is a rare manifestation of extrapulmonary TB. This disease entity can be overlooked, and its early diagnosis is difficult because of its variable manifestations and lack of specific symptoms. METHODS: The clinical and diagnostic features, treatment, and outcome of patients with abdominal TB at a major hospital in southeastern Taiwan from January 1987 to December 2006 were investigated. RESULTS: Twenty-one patients with abdominal TB identified during the 20-year period were included. A predominance of male (13/21, 61.9%) and Taiwanese aborigine (15/21, 71.4%) patients was noted. Middle-aged (30-50 years) patients with alcoholic liver cirrhosis had the highest risk. Common presenting features included abdominal pain (18/21, 85.7%), fever (16/21, 76.2%), ascites (13/21, 61.9%), and weight loss (12/21, 57.3%). The mean time to reach a diagnosis was 48 +/- 10 days. Tuberculous peritonitis was noted in 11 patients, with a high correlation with liver cirrhosis (p = 0.0237, Fisher's exact test). The other patients were diagnosed with TB of the gastrointestinal tract (n = 6), urinary tract (n = 2), and pelvis (n = 2). Abdominal sonography and abdominal computed tomography were helpful for diagnosis, by revealing ascites and thickening of the peritoneum. Pulmonary involvement was noted in 11 patients. Most of the patients (16/21, 76.2%) improved with anti-tuberculosis therapy, and five patients died from sepsis and respiratory failure. CONCLUSION: Abdominal TB is a rare manifestation of extrapulmonary TB, even in southeastern Taiwan where TB is prevalent. Delay in diagnosis is common and abdominal TB should be included in the differential diagnosis of middle-aged aborigine men with alcoholic liver cirrhosis and peritonitis. PMID- 19293035 TI - Delayed suspicion, treatment and isolation of tuberculosis patients in pulmonology/infectious diseases and non-pulmonology/infectious diseases wards. AB - BACKGROUND/PURPOSE: Delayed diagnosis and isolation increases the risk of nosocomial transmission of tuberculosis (TB). To assess the risk of delayed management of TB, we analyzed the risk factors of prolonged delay in isolation of smear-positive TB patients in pulmonology/infectious diseases and other wards in a tertiary teaching hospital. METHODS: We enrolled smear-positive TB patients aged > 16 years with delayed respiratory isolation following hospitalization. Medical records were reviewed retrospectively. Time intervals between admission, order of sputum acid-fast staining, initiation of anti-tuberculous treatment and isolation were compared between pulmonology/infectious diseases wards (PIWs) and other wards. Risk factors were analyzed in patients with prolonged isolation delay of > 7 days in individual groups. RESULTS: Isolation was delayed in 191 (73.7%) of 259 hospitalized smear-positive TB patients. Median suspicion, treatment and isolation delays were 0, 3 and 4 days in PIWs and 1, 5 and 7 days in other wards. For patients admitted to non-PIWs, atypical chest radiographs, symptoms without dyspnea or not being admitted from the emergency department (ED) were risk factors for prolonged isolation delay exceeding 7 days. The only risk factor for delayed isolation in patients admitted to PIWs was age >or= 70 years. CONCLUSION: Delays in suspicion, treatment and isolation of TB patients were longer in non-PIWs. Clinicians should be alert to those admitted to non-PIWs with atypical chest radiographs, atypical symptoms, or not admitted from the ED. PMID- 19293036 TI - Clinical implications of alpha-fetoprotein in chronic hepatitis C. AB - BACKGROUND/PURPOSE: Chronic hepatitis C (CHC) shows a significant association with cirrhosis and hepatocellular carcinoma (HCC). Alpha-fetoprotein (AFP) is important in the diagnosis of HCC, but elevated AFP levels have also been observed in CHC without HCC. We evaluated the clinical correlation between elevated AFP levels and CHC. METHODS: From April 1999 to November 2004, 654 CHC patients with no evidence of HCC from imaging studies were collected by chart review. RESULTS: The prevalence of elevated AFP levels (>or= 15 ng/mL) was 23.9%. Univariate analysis revealed that age, histological activity index (HAI) fibrosis score of 3/4, HAI inflammation score >or= 7, aspartate aminotransferase (AST) and alanine transaminase (ALT) levels, AST/ALT ratio, and total bilirubin level were associated with elevated AFP levels. Multivariate analysis revealed that age (>or= 55 vs. < 55 years), HAI inflammation score (>or= 7 vs. < 7), ALT (> 150 vs. 150 x 109 cells/L) were associated with elevated AFP levels. Multivariate analysis also revealed that hepatitis C virus (HCV) genotype 1b, platelet count 80 U/L and AFP >or= 6 ng/mL were associated with advanced fibrosis. Using a cut off AFP level of >or= 6.0 ng/mL, the sensitivity and specificity of diagnosing fibrosis score 3/4 was 74.3% and 68.4%, respectively. Using a cut-off AFP level of >or= 15.0 ng/mL, the sensitivity and specificity of diagnosing fibrosis score 3/4 was 35.7% and 91.1%, respectively. CONCLUSION: Elevated AFP levels were observed in 23.9% of patients with CHC. Elevated AFP levels correlated positively with age, HAI inflammation score, ALT elevation, and thrombocytopenia. In addition, HCV genotype 1b, thrombocytopenia, AST elevation, and AFP level >or= 6 ng/mL were associated with advanced fibrosis. PMID- 19293037 TI - Repeated pneumonia severity index measurement after admission increases its predictive value for mortality in severe community-acquired pneumonia. AB - BACKGROUND/PURPOSE: Severe community-acquired pneumonia (CAP) is associated with high hospital mortality, and accurate assessment of patients is important for supporting clinical decision making. The Pneumonia Severity Index (PSI) is a good tool for predicting disease severity, especially in the low-risk group of patients with CAP. We investigated whether the change in PSI measurement after admission could identify patients at high risk of mortality from CAP. METHODS: We prospectively studied 250 inpatients with CAP. PSI was measured at admission and 72 hours later at a tertiary referral medical center from May 2005 to February 2006. The initial and repeated PSI results were compared. Hospital mortality was used as the outcome measure. RESULTS: Initial PSI in high-risk patients (PSI class > IV) had a low specificity (37%), and a low positive predictive value (PPV) (17%). Increased repeated PSI score, as compared with initial score, was associated with an increased mortality rate (from 7.8% to 33.3% in class IV, and 25.3% to 53.3% in class V; p < 0.0001), and improved the predictive value, with 94% specificity and a PPV of 46% for mortality in high-risk patients. CONCLUSION: Increased PSI score, 72 hours after admission, for patients with CAP improved the predictive value of PSI score and more accurately identified patients with a high risk of mortality. PMID- 19293038 TI - Impact of silent infarction on the outcome of stroke patients. AB - BACKGROUND/PURPOSE: Silent infarcts (SIs) are commonly found on brain computed tomography (CT) or magnetic resonance imaging (MRI) among elderly subjects, but their risk factors and impact on outcome in stroke patients are unknown. We evaluated the prevalence, risk factors and impact of SIs on the outcome of patients admitted with first-ever ischemic stroke or transient ischemic attack (TIA). METHODS: A prospective study of 446 patients admitted consecutively to the neurology service with a diagnosis of TIA or stroke between July 2003 and June 2005, including 226 without any history of prior TIA or stroke. All patients underwent brain CT on the day of admission to the hospital. Risk factors analyzed included age, history of hypertension, diabetes mellitus, cardiovascular disease or stroke, smoking habit and alcohol use. Cholesterol and triglyceride levels were measured on the second day of admission. We monitored these patients for 24 months after stroke onset. RESULTS: The frequency of SIs among the 226 patients with first-ever stroke or TIA was 20%. Most of the SIs were small and deep. Small artery disease was more frequently observed in patients with SIs. Age, hypertension, diabetes mellitus, hypercholesterolemia, hypertriglyceridemia, alcohol use, smoking habits and atrial fibrillation did not significantly differ between patients with SIs and those without SIs. During the 24-month follow-up period, the frequency of recurrent stroke was higher in patients with SIs than those without SIs. The mortality rate was higher in patients without SIs than those with SIs. The interval from stroke onset to rehospitalization was shorter in patients without SIs than in those with SIs. CONCLUSION: The study showed a higher frequency of small artery disease in patients with SIs. First-ever stroke patients with SIs should be considered at high risk for recurrent stroke. PMID- 19293039 TI - Predicting factors in the last week of survival in elderly patients with terminal cancer: a prospective study in southern Taiwan. AB - BACKGROUND/PURPOSE: The proportion of elderly persons among terminal cancer patients is around 60% in Taiwan. Prediction of the last week of survival can be significant in palliative care units for planning patient management, fulfilling patients' wishes at the end of life, and meeting the needs of relatives. We aimed to find out the predictive factors for the last week of survival in elderly terminal cancer patients. METHODS: We prospectively observed items based on previous research that we anticipated might influence survival among 459 consecutive elderly patients. Within 24 hours of admission, we collected data including clinical symptoms and signs, demographic information, and biochemical test results. Multivariate logistic regression analysis was performed to identify factors associated with patient mortality within 7 days. RESULTS: Multivariate analysis indicated that the following factors were predictive of death: higher score of Eastern Cooperative Oncology Group (ECOG) performance status (OR, 2.018; p < 0.001), primary liver cancer (OR, 1.968; p = 0.024), male gender (OR, 2.885; p < 0.001), lower extremity muscle power (OR, 0.722; p = 0.0266), lower systolic blood pressure (OR, 0.985; p = 0.011), higher heart rate (OR, 1.017; p = 0.016), higher hemoglobin (OR, 1.216; p = 0.003), and higher blood urea nitrogen (BUN) (OR, 1.028; p < 0.001). CONCLUSION: We propose that the probability of survival of less than a week can be predicted by our formula. This formula, which includes factors of demographic information (male gender, and presence of liver cancer), clinical signs (lower systolic blood pressure, higher heart rate, and lower mean extremity muscle power), and biochemical tests (elevated BUN, and higher hemoglobin), may help improve survival prediction in terminal elderly cancer patients. PMID- 19293040 TI - Increased connective tissue growth factor expression in a rat model of chronic heart allograft rejection. AB - BACKGROUND/PURPOSE: Chronic rejection limits the long-term success of cardiac transplantation and the underlying cause of the disease is unknown. Connective tissue growth factor (CTGF) is considered as a mitogenic and chemotactic factor for fibroblasts, and is associated with cell proliferation and collagen synthesis. We evaluated the expression of CTGF in a rat model of heart allograft chronic rejection. METHODS: Intra-abdominal heterotopic heart transplantation was performed from 20 Wistar rats to 20 Sprague-Dawley (SD) rats that received cyclosporine, mycophenolate mofetil and methylprednisolone as immunosuppression. Ten heart allografts were explanted at 2 and 8 weeks postoperatively for analysis of morphologic changes. The hearts from 10 normal Wistar rats served as a control group. Coronary artery density, luminal loss of myocardial coronary arteries, and myocardial fibrosis were measured. The expression of CTGF was studied by immunohistochemistry. Correlation between CTGF expression and development of cardiac allograft vasculopathy (CAV) or fibrosis was studied. RESULTS: Allografts harvested at 8 weeks postoperatively showed more coronary intimal proliferation, fibrosis and CTGF expression compared with the 2-week allografts (p < 0.05) and the controls (p < 0.01), but the coronary artery density was lower than in the control group (p < 0.05). However, the control group showed negligible CTGF expression. There were strong negative correlations between the gray value of CTGF protein expression and cardiac fibrosis and coronary intimal occlusion (r = 0.734, -0.713; p < 0.01), which demonstrated that CTGF protein expression was positively correlated with cardiac fibrosis and coronary intimal occlusion. CONCLUSION: CTGF is expressed in cardiomyocytes in CAV. Increased expression of CTGF in cardiac allografts is associated with development of CAV and fibrosis formation. PMID- 19293041 TI - Meconium pseudocyst: a classical and successfully treated case. AB - Meconium peritonitis with pseudocyst formation is rare and can be lethal. We report a newborn infant with antenatal sonographic diagnosis of bowel dilatation and giant meconium pseudocyst. Postnatal presentation included a palpable abdominal mass and abdominal distention. Abdominal X-ray revealed a huge egg shell calcified mass containing air-fluid level. Postnatal sonography revealed a cystic mass with air-fluid level and acoustic shadows. Computed tomography showed a giant communicating cyst with egg-shell calcification, which compressed the other intestinal loops to the posterior peritoneal cavity. Elective laparotomy was performed, and distal ileal atresia with sealed proximal perforation and a giant meconium pseudocyst were found. Resection of the involved small bowel, including the giant pseudocyst, followed by primary end-to-end anastomosis was performed smoothly. The postoperative course was uneventful. Advances in perinatal intensive care mean that neonates can be operated upon under stable rather than critical conditions, and elective rather than emergency laparotomy, and primary anastomosis of the intestine rather than staged enterostomy can be performed. PMID- 19293042 TI - Pyogenic liver abscess with Prevotella species and Fusobacterium necrophorum as causative pathogens in an immunocompetent patient. AB - Pyogenic liver abscess of odontogenic origin in an immunocompetent patient is extremely rare. We report an immunocompetent 25-year-old male hepatitis B carrier with severe dental disease that led to the development of liver abscess. A periapical abscess in the upper left molar area was seen on his dental X-ray. Two sets of blood cultures grew Prevotella species, bacteria that are commonly found inside the oral cavity. Bacterial culture of the liver abscess drainage sample grew both Prevotella and Fusobacterium necrophorum. This led to our diagnosis of pyogenic liver abscess of dental origin, since we found no other source of infection in our patient except for his dental disease. After antibiotic therapy with drainage, abdominal sonography showed resolution of the abscess. The diseased teeth were also extracted. During 1 year of follow-up, there was no sign of abscess recurrence. A diagnosis of pyogenic liver abscess necessitates a complete evaluation to rule out possible biliary, colonic or other associated diseases. However, when a liver abscess is thought to be cryptogenic, we also recommend a careful dental examination to help identify the source of infection. PMID- 19293043 TI - Spontaneous spinal and intracranial subdural hematoma. AB - Subdural hematoma (SDH) of the spine following intracranial hemorrhage is extremely rare. We present a 35-year-old woman who suffered from headache and dizziness initially, and then lower back pain, lower limb weakness and paraparesis gradually developed within 1-2 weeks. Magnetic resonance imaging revealed intracranial and spinal SDH. No vascular abnormality was seen by brain and spinal angiography. Platelet count, prothrombin time, activated partial thromboplastin time, and inflammatory markers, including C-reactive protein, were normal. A diagnosis of spontaneous spinal and intracranial SDH was then confirmed surgically. Postoperative recovery was uneventful. PMID- 19293044 TI - Non-alcoholic fatty liver disease and antiviral therapy in hepatitis C. PMID- 19293047 TI - Beware the node: the more we know, the less aggressive we are. PMID- 19293048 TI - What a strange thing it is to live. PMID- 19293049 TI - Biology of BMP signalling and cancer. AB - In recent years, it has been proposed that tumours are not homogeneous but composed of several cellular types like normal tissues. A cellular subtype, which is though to be the origin of tumours as well as their malignant properties (i.e., capacity for regrowth and metastasis), are the cancer stem cells (CSCs). CSCs, like normal stem cells, have a nearly unlimited capacity to self-renew and to proliferate so that are responsible, besides their same auto-perpetuation giving rise to the features previously depicted, also for the generation of the bulk of more differentiated cells in tumour. The altered behaviour of CSCs may be caused by the malfunction of a number of signalling pathways involved in normal embryonic development and in tissue homeostasis in adulthood. Among these signalling pathways are Wnt, Hedgehog, Notch and BMP pathways. In this review, we will focus on the study of molecular aspects of BMP signalling as well as its involvement in cancer. PMID- 19293050 TI - Tumoral angiogenesis and breast cancer. AB - Breast cancer (BC) is the most common neoplasm in women in Western countries. Tumoral angiogenesis (TA) is essential for the growth and spread of BC cells. There are at least 6 different angiogenic growth factors associated with TA in BC. The major mediator of TA is vascular endothelial growth factor (VEGF), a homodimeric heparin-binding glycoprotein. VEGF signals through VEGF receptor-2 (VEGFR-2), the major VEGF signalling receptor that mediates sprouting angiogenesis. Recently, different antiangiogenic agents have shown efficacy in the treatment of advanced BC. Bevacizumab, a humanised monoclonal antibody against VEGF, in combination with taxanes improves progression-free survival and overall response rate in first-line therapy. Other new antiangiogenic agents, called multi-kinase inhibitors (sunitinib and pazopanib), are under investigation. Finally, a schedule of treatment called metronomic chemotherapy, with antiangiogenic activity, has also demonstrated efficacy in the treatment of advanced BC. PMID- 19293051 TI - Use of radionuclides in cancer research and treatment. AB - Cancer occurs as a result of misregulation of cell growth, which appears to be a consequence of alteration in the function of oncogenes and tumour suppressor genes. Ionising radiation has been used, since the discovery of X-rays in 1896 by Roentgen, both in cancer research and treatment of the disease. The main purpose of cancer research is to understand the molecular alterations involved in the development and progression of the disease in order to improve diagnosis and develop personalised therapies, by focusing on the features of the tumoral cell and the biological events associated to carcinogenesis. Radioisotopic techniques have been used routinely for in vitro research in the molecular and cellular biology of cancer for more than 20 years and are in the process of being substituted by alternative non-radioactive techniques. However in vivo techniques such as irradiation of cells in culture and/or experimental animal models and radioactive labelling are in development, due in part to advances in molecular imaging technologies. The objective of this review is to analyse in an integrative way the applications of ionising radiation in cancer research and therapy. It had been divided into two parts. The first one will approach the techniques applied to cancer research and the second will summarise how ionising radiation is applied to the treatment of neoplastic disease. PMID- 19293052 TI - New approaches to female fertility preservation. AB - Survival rates have been improved in young cancer patients due to advances in oncologic treatments. It is necessary to focus on the consequences of cancer treatments and try to elude or at least prevent them. Menopause and infertility are two of the main causes for concern to those patients who survive cancer. When a solution is offered to these patients, their quality of life and self-esteem can be dramatically increased after therapy. Results observed in vitrified oocytes are similar to those achieved with fresh oocytes, giving this method great potential. Recommendations on fertility preservation should be focused on the oocyte vitrification method, which should be considered as the first option. Limitations of this approach are the need for ovarian stimulation, which means a 2-3 week period prior to chemotherapy and the possibility of high oestradiol levels when tumours are hormone dependent. PMID- 19293053 TI - The EORTC Quality of Life Questionnaire for patients with prostate cancer: EORTC QLQ-PR25. Validation study for Spanish patients. AB - OBJECTIVES: The EORTC Quality of Life (QL) Group has developed a questionnaire, the EORTC QLQ-PR25, for evaluating QL in prostate cancer. The aim of this study is to assess the psychometric properties of the EORTC QLQPR25 when applied to a sample of Spanish patients. MATERIALS AND METHODS: One hundred and thirty-seven prostate cancer patients with localised disease who started radiotherapy with radical intention combined with or without hormonotherapy prospectively completed the EORTC QLQ-C30 and EORTC QLQ-PR25 questionnaires three times: on the first and last day of radiotherapy and in the follow-up period. Psychometric evaluation of the questionnaires' structure, reliability and validity was conducted. RESULTS: Multitrait scaling analysis showed that many of the item-scale correlation coefficients met the standards of convergent and discriminant validity. Exceptions appeared mainly in the scales for bowel symptoms and for hormonal- treatment-related symptoms. Cronbach's coefficients of the scales were good (0.72 0.86) for the urinary symptoms and sexual function scales but they were lower (<0.70) for the bowel and hormonal treatment scales. Most scales of the EORTC QLQ PR25 had low to moderate intercorrelations. Correlations between the scales of the QLQ-C30 and the module were generally low. Group comparison analyses showed better QL in patients with higher Performance Status. Changes in QL appeared throughout the measurements. These were in line with the treatment process. CONCLUSIONS: The EORTC QLQ-PR25 was a reliable and valid instrument when applied to a sample of Spanish prostate cancer patients. These results are in line with those of the EORTC validation study. PMID- 19293054 TI - Predictive factors for non-sentinel lymph node involvement in breast cancer patients with a positive sentinel node: should we consider sentinel node-related factors? AB - INTRODUCTION: Sentinel node (SN) biopsy to predict axillary involvement in breast cancer patients is a common practice. After a positive SN, additional metastases are present in an unpredictable percentage, as variable as 13-66%, of axillary clearances. Our aim is to define variables associated with non-sentinel node (NSN) metastases and to determine the predictive value of a derived clinical decision rule. METHODS: A consecutive series of patients with a positive SN submitted to SN biopsy plus axillary dissection was evaluated from June 1999 to December 2004 (n=143). Patient-, tumour- and SN-related variables were analysed in relation to the presence of additional metastasis. Univariate and multivariate analyses were done and predictive values of a clinical decision rule based on significant variables were estimated. RESULTS: A total of 66 patients had metastasis in non-sentinel axillary nodes. No significant differences were present between this group and those with only the SN metastasised. Significant and independent association was found between NSN positivity and increasing tumour size, the presence of multifocality and the presence of peritumoral lymph channel invasion. CONCLUSIONS: A first derivation of a simple rule based on tumour-related variables concurs to define the presence of NSN metastasis. Care should be taken when including SNrelated variables in these algorithms. PMID- 19293055 TI - Feasibility of adequate resectable rectal cancer treatment in a third-level hospital. AB - PURPOSE: The aim of this study was to determine the feasibility, concerning compliance to protocol and recommended clinical practice guidelines, as well as efficacy results of multidisciplinary treatment (surgery, radiotherapy and chemotherapy) of resectable rectal cancer in a third-level hospital devoid of radiotherapy and clinical oncology units. PATIENTS AND METHODS: A retrospective, single-institution analysis was completed for 45 consecutive patients diagnosed with resectable rectal cancer who entered an officially proposed multidisciplinary treatment protocol from October 1998 to September 2003. Adequacy of patient inclusion, according to clinical stage, was reviewed. Neoadjuvant radiotherapy schedule, surgery procedures and adjuvant chemotherapy indication were assessed. All treatment time intervals were analysed. Finally, efficacy results are discussed and contextualised by comparison with results of clinical trials which support this treatment strategy. RESULTS: According to an independent board review, 3 patients (6.7%) with stage I rectal cancer, 31 patients (68.9%) with stage II and 11 patients (24.4%) with stage III rectal cancer were included. Radiotherapy dosage, volume and schedule were as planned. Median time from diagnosis to start of radiotherapy was 26.36 days (24.26- 28.57; CI 95%). Median duration of radiotherapy was 6.00 days (5.56-6.44; CI 95%). Median time from start of radiotherapy to surgery was 15.67 days (14.47-16.87; CI 95%). Median time from completion of radiotherapy to surgery was 10.67 days (9.53 11.81; CI 95%). Most of the patients underwent low anterior resection [23 patients (51.2%)] and abdominoperineal resection [16 patients (35.6%)]. Correlation between clinical and pathologic staging was as expected. Twenty-nine patients (64.4%) of the 45 that were initially included started adjuvant chemotherapy. A statistically significant relationship between pathologic stage (grouped I-II vs. III) and the use of adjuvant chemotherapy was found (p=0.033; chi-square test). Radiotherapy- and chemotherapy-induced toxicity did not differ from that previously reported. With a median follow-up of 65.46 months, a total of 10 recurrences have been diagnosed, all of them in stage III patients. Overall survival rate at five years was 76% for the complete population included. CONCLUSION: Multidisciplinary treatment of resectable rectal cancer in a third level hospital is feasible. Although efficacy results are comparable to those previously reported in the literature, further improvements in clinical staging as well as in adjuvant chemotherapy indication are desirable. PMID- 19293056 TI - Results of R0 surgery with D2 lymphadenectomy for the treatment of localised gastric cancer. AB - INTRODUCTION: Surgical excision is the best therapeutic option for gastric cancer, provided it is performed with curative intent as R0 surgery. And, according to many authors, D2 lymphadenectomy may be performed with acceptable morbidity and mortality rates. MATERIALS AND METHODS: A prospective study was conducted on a series of 126 consecutive cases of gastric cancer treated with gastrectomy and D2 lymphadenectomy. A R0 resection was done in 99 cases (78.6%). RESULTS: Total gastrectomy was performed in 70 patients and subtotal gastrectomy in 29. The mean number of lymph nodes removed was 32.5 per patient. Suture dehiscence occurred in 3 patients (in one of them in the esophago-jejunal anastomosis). Hospital mortality was 2%. Twenty-six recurrences were detected after a median follow-up of 73.6 months. Five-year actuarial survival was 65%. Five-year survival of patients with positive lymph nodes at the N2 level was 26.5%. CONCLUSIONS: Gastrectomy with D2 lymphadenectomy may be performed with low morbidity and mortality. R0 resection allows acceptable survival rates to be achieved. There is even a group of patients with invaded lymph nodes at the N2 level surviving at 5 years. It appears to be very important that this surgery is performed by specialised surgeons. PMID- 19293057 TI - Acute ischaemic cerebrovascular attack secondary to infusional 5-fluoruracil and cisplatin in a patient with advanced gastric cancer. AB - Acute ischaemic cerebrovascular attack may be an underreported complication related to chemotherapy. We report here the case of a patient with acute ischaemic cerebrovascular attack, immediately after administration of a first cycle of chemotherapy based on 5-fluoruracil and cisplatin. PMID- 19293058 TI - Primary non-Hodgkin lymphoma and invasive ductal carcinoma in the same breast: a rare case report. AB - Primary lymphoma of the breast is an unusual clinical entity. The coexistence in the same breast of an invasive ductal carcinoma is even rarer. We report a 69 year old woman referred for further evaluation of a palpable mass in her right breast. She was diagnosed and treated for simultaneous primary lymphoma and invasive ductal carcinoma. Primary breast lymphoma should always be considered in the differential diagnosis of breast masses. The presence of both malignancies presents a challenge in treatment decisions. PMID- 19293059 TI - [Physical activity in prevention and treatment of diabetes]. AB - Effects and mechanisms of a single bout of physical load and of the regular exercise (training) on the carbohydrate and fat metabolism are reviewed. During exercise and in the following couple of hours sugar utilization improves, "activity functions like insulin". Proper exercise contributes to using up the fat reserves, and slimmer body, fat reduction can be maintained by the combination of exercise and diet-control. Instructions in "exercise for everybody" and particularly for both type diabetics are detailed. In prevention of cardio-metabolic pathologies the most important measure were avoiding the overweight state by rational alimentation and regular physical activity. PMID- 19293060 TI - [Mood disorders in patients with chronic kidney disease: significance, etiology and prevalence of depression]. AB - Due to the rapidly increasing number of end-stage renal disease patients and the high costs of their treatment, all the aspects of kidney disease that may significantly affect clinical outcome (quality of life mortality) deserve increasing attention. It has been established and accepted that in addition to clinical/somatic factors, also psycho-social factors, including depression, may have a significant impact on the clinical outcome of chronic diseases. Depression is considered to be one of the most prevalent mental health problems in patients with chronic kidney disease. In spite of this fact, there are only few studies on the prevalence, diagnosis and treatment of depression in this population using accurate, well defined diagnostic criteria and appropriate epidemiologic methods. In the last decades we have experienced a significant improvement in the quality and effectiveness of the therapeutic options for chronic kidney disease, but mortality is still very high in this population. Our review provides an overview of the literature regarding the prevalence and etiology of depression, and calls the attention to the interrelation among depression, quality of life and mortality. The second part of our paper to be published later will survey the specific diagnostic and therapeutic features of depression in chronic kidney disease patients. PMID- 19293061 TI - [Role of myofibroblast in inflammatory bowel disease and tumor genesis]. AB - Stroma cells with the microenvironment around them have primary role in the regulation of inflammation processes, conformation of tumors and development of metastasis. Myofibroblasts have essential role in inflammation processes and in the regeneration of encompassed tissue. Molecules produced by them operate the cells of the immune system and effect the proliferation of epithelium. Tumors can activate myofibroblasts which can lead to uncontrolled epithelium proliferation across production of changed and increased regulation ligands (such as cytokines, chemokines, chemotactic and other growth factors) and activation of stem cell. This process could lead piling of uncontrolled epithelial cells and can impact later on conformation of tumors. In this study we present an overview about of myofibroblasts and their roles in inflammation and neoplastic processes. PMID- 19293062 TI - [Posterior circumflex humeral artery perforator flap (PCHAP-flap) in axillary scar release]. AB - Scar contracture after burn injury is frequent in the axillary region. If conservative treatment of scars limiting the range of motion of the shoulder joint fails, surgical correction is needed. The authors performed an early reconstruction in a case with unstable scar formation and limited range of motion after burn injury to the axillary region, using the posterior circumflex humeral artery perforator flap. In their opinion, the flap (beside its conventional use as a free flap) may be useful as an island flap in the reconstruction of the axillary region, e. g. for axillary scar release. PMID- 19293063 TI - [Perspectives of inhibition of multidrug resistance during cancer chemotherapy, in vitro and in vivo experiments]. AB - The development of pharmacological agents able to counteract the mechanisms of multidrug resistance in oncology has remained a major goal for the past ten years. Our purpose was to find multidrug resistance reversal agents less toxic than verapamil among various synthetic compounds: cinnamylidene ketones; 1,4 dihydropyridines; phenothiazines; heat shock 90 inhibitor peptides; betti base derivative of tylosin and among some naturally occurring plant derived jatrophane and lathyrane-type diterpenes. The first part of this thesis presents the inhibition of multidrug resistance through inhibition of the P-glycoprotein efflux pump in various cell lines. In general, the newly identified multidrug resistance modifiers were able to enhance the antiproliferative activity of selected anticancer drugs in a synergistic or additive way in in vitro experiments. The in vitro activity of betti base derivative of tylosin was confirmed by further in vivo efficacy studies in DBA/2 mice. As an alternative way of antitumor effect, apoptosis inductions of resistance modifiers were studied. The substituted dihydropyridine 13 was the most promising apoptosis inducer on mouse lymphoma cells. Human cytomegalovirus was used in a modified in vitro model for characterizing lathyrane compounds with antipromotion effect on human lung cancer cells. All the six macrocyclic lathyrane-type diterpenoids reduced the promotion in vitro , except latilagascene D, decreased IE-antigen expression of cytomegalovirus to prevent progression of tumor malignancy. PMID- 19293067 TI - Summaries for patients. What aspirin dose is safest and most effective for preventing heart disease? PMID- 19293068 TI - Summaries for patients. Aspirin for the prevention of cardiovascular disease: U.S. Preventive Services Task Force recommendation statement. PMID- 19293069 TI - B-type natriuretic peptide testing, clinical outcomes, and health services use in emergency department patients with dyspnea: a randomized trial. AB - BACKGROUND: B-type natriuretic peptide (BNP) is used to diagnose heart failure, but the effects of using the test on all dyspneic patients is uncertain. OBJECTIVE: To assess whether BNP testing alters clinical outcomes and health services use of acutely dyspneic patients. DESIGN: Randomized, single-blind study. Patients were assigned to a treatment group through randomized numbers in a sealed envelope. Patients were blinded to the intervention, but clinicians and those who assessed trial outcomes were not. SETTING: 2 Australian teaching hospital emergency departments. PATIENTS: 612 consecutive patients who presented with acute severe dyspnea from August 2005 to March 2007. INTERVENTION: BNP testing (n = 306) or no testing (n = 306). MEASUREMENTS: Admission rates, length of stay, and emergency department medications (primary outcomes); mortality and readmission rates (secondary outcomes). RESULTS: There were no between-group differences in hospital admission rates (85.6% [BNP group] vs. 86.6% [control group]; difference, -1.0 percentage point [95% CI, -6.5 to 4.5 percentage points]; P = 0.73), length of admission (median, 4.4 days [interquartile range, 2 to 9 days] vs. 5.0 days [interquartile range, 2 to 9 days]; P = 0.94), or management of patients in the emergency department. Test discrimination was good (area under the receiver-operating characteristic curve, 0.87 [CI, 0.83 to 0.91]). Adverse events were not measured. LIMITATION: Most patients were very short of breath and required hospitalization; the findings might not apply for evaluating patients with milder degrees of breathlessness. CONCLUSION: Measurement of BNP in all emergency department patients with severe shortness of breath had no apparent effects on clinical outcomes or use of health services. The findings do not support routine use of BNP testing in all severely dyspneic patients in the emergency department. PRIMARY FUNDING SOURCE: Janssen-Cilag. PMID- 19293070 TI - Trajectories of life-space mobility after hospitalization. AB - BACKGROUND: Life space is a measure of where a person goes, the frequency of going there, and the dependency in getting there. It may be a more accurate measure of mobility in older adults because it reflects participation in society as well as physical ability. OBJECTIVE: To assess effects of hospitalization on life space in older adults, and to compare life-space trajectories associated with surgical and nonsurgical hospitalizations. DESIGN: Prospective observational study. SETTING: Central Alabama. PARTICIPANTS: 687 community-dwelling Medicare beneficiaries at least 65 years of age with surgical (n = 44), nonsurgical (n = 167), or no (n = 476) hospitalizations. MEASUREMENTS: Life-Space Assessment (LSA) scores before and after hospitalization (range, 0 to 120; higher scores reflect greater mobility). RESULTS: Mean age of participants was 74.6 years (SD, 6.3). Fifty percent were black, and 46% were male. Before hospitalization, adjusted LSA scores were similar in participants with surgical and nonsurgical admissions. Life-space assessment scores decreased in both groups immediately after hospitalization; however, participants with surgical hospitalizations had a greater decrease in scores (12.1 more points [95% CI, 3.6 to 20.7 points]; P = 0.005) than those with nonsurgical hospitalizations. However, participants with surgical hospitalizations recovered more rapidly over time (gain of 4.7 more points [CI, 2.0 to 7.4 points] per ln [week after discharge]; P < 0.001). Score recovery for participants with nonsurgical hospitalizations did not significantly differ from the null (average recovery, 0.7 points [CI, -0.6 to 1.9 points] per ln [week after discharge]). LIMITATION: Life space immediately before and after hospitalization was self-reported, often after hospital discharge. CONCLUSION: Hospitalization decreases life space in older adults. Surgical hospitalizations are associated with immediate marked life-space declines followed by rapid recovery, in contrast to nonsurgical hospitalizations, which are associated with more modest immediate declines and little evidence of recovery after several years of follow-up. PRIMARY FUNDING SOURCE: National Institute on Aging. PMID- 19293071 TI - Aspirin to prevent cardiovascular disease: the association of aspirin dose and clopidogrel with thrombosis and bleeding. AB - BACKGROUND: The optimal aspirin dose for the prevention of cardiovascular events remains controversial. OBJECTIVE: To assess the incidence of and risk factors for adverse clinical outcomes by investigator-determined aspirin dose in a primary prevention trial. DESIGN: Post hoc observational analyses of data from a double blind, placebo-controlled, randomized trial. SETTING: Outpatient. PATIENTS: 15 595 patients with cardiovascular disease or multiple risk factors. INTERVENTION: Clopidogrel, 75 mg/d, or placebo, with aspirin, 75 to 162 mg/d, as selected by the investigators. MEASUREMENTS: Incidence of the composite outcome of myocardial infarction, stroke, or cardiovascular death (efficacy end point), and incidence of severe or life-threatening bleeding (safety end point), at a median of 28 months (interquartile range, 23 to 31 months) of follow-up. RESULTS: Daily aspirin doses were categorized as less than 100 mg (75 or 81 mg) (n = 7180), 100 mg (n = 4961), and greater than 100 mg (150 or 162 mg) (n = 3454). The hazard of the primary efficacy end point was the same regardless of dose (adjusted hazard ratio, 0.95 [95% CI, 0.80 to 1.13] for 100 mg vs. less than 100 mg, and 1.0 [CI, 0.85 to 1.18] for greater than 100 mg vs. less than 100 mg). The hazard of the primary safety end point also did not depend on dose (adjusted hazard ratio, 0.85 [CI, 0.57 to 1.26] for 100 mg vs. less than 100 mg and 1.05 [CI, 0.74 to 1.48] for greater than 100 mg vs. less than 100 mg). In patients also receiving clopidogrel, daily aspirin doses greater than 100 mg seemed to be non statistically significantly associated with reduced efficacy (adjusted hazard ratio, 1.16 [CI, 0.93 to 1.44]) and increased harm (adjusted hazard ratio, 1.30 [CI, 0.83 to 2.04]). LIMITATION: The analysis was post hoc, and aspirin use was not randomized or blinded. CONCLUSION: Daily aspirin doses of 100 mg or greater were associated with no clear benefit in patients taking aspirin only and possibly with harm in patients taking clopidogrel. Daily doses of 75 to 81 mg may optimize efficacy and safety for patients requiring aspirin for long-term prevention, especially for those receiving dual antiplatelet therapy. PRIMARY FUNDING SOURCE: None. PMID- 19293072 TI - Aspirin for the prevention of cardiovascular disease: U.S. Preventive Services Task Force recommendation statement. AB - DESCRIPTION: Update of the 2002 U.S. Preventive Services Task Force (USPSTF) recommendation about the use of aspirin for the prevention of coronary heart disease. METHODS: Review of the literature since 2002, focusing on new evidence on the benefits and harms of aspirin for the primary prevention of cardiovascular disease, including myocardial infarction and stroke. The new evidence was reviewed and synthesized according to sex. RECOMMENDATIONS: Encourage men age 45 to 79 years to use aspirin when the potential benefit of a reduction in myocardial infarctions outweighs the potential harm of an increase in gastrointestinal hemorrhage. (A recommendation) Encourage women age 55 to 79 years to use aspirin when the potential benefit of a reduction in ischemic strokes outweighs the potential harm of an increase in gastrointestinal hemorrhage. (A recommendation) Evidence is insufficient to assess the balance of benefits and harms of aspirin for cardiovascular disease prevention in men and women 80 years or older. (I statement) Do not encourage aspirin use for cardiovascular disease prevention in women younger than 55 years and in men younger than 45 years. (D recommendation). PMID- 19293073 TI - Aspirin for the primary prevention of cardiovascular events: an update of the evidence for the U.S. Preventive Services Task Force. AB - BACKGROUND: Coronary heart disease and cerebrovascular disease are leading causes of death in the United States. In 2002, the U.S. Preventive Services Task Force (USPSTF) strongly recommended that clinicians discuss aspirin with adults who are at increased risk for coronary heart disease. PURPOSE: To determine the benefits and harms of taking aspirin for the primary prevention of myocardial infarctions, strokes, and death. DATA SOURCES: MEDLINE and Cochrane Library (search dates, 1 January 2001 to 28 August 2008), recent systematic reviews, reference lists of retrieved articles, and suggestions from experts. STUDY SELECTION: English language randomized, controlled trials (RCTs); case-control studies; meta analyses; and systematic reviews of aspirin versus control for the primary prevention of cardiovascular disease (CVD) were selected to answer the following questions: Does aspirin decrease coronary heart events, strokes, death from coronary heart events or stroke, or all-cause mortality in adults without known CVD? Does aspirin increase gastrointestinal bleeding or hemorrhagic strokes? DATA EXTRACTION: All studies were reviewed, abstracted, and rated for quality by using predefined USPSTF criteria. DATA SYNTHESIS: New evidence from 1 good-quality RCT, 1 good-quality meta-analysis, and 2 fair-quality subanalyses of RCTs demonstrates that aspirin use reduces the number of CVD events in patients without known CVD. Men in these studies experienced fewer myocardial infarctions and women experienced fewer ischemic strokes. Aspirin does not seem to affect CVD mortality or all-cause mortality in either men or women. The use of aspirin for primary prevention increases the risk for major bleeding events, primarily gastrointestinal bleeding events, in both men and women. Men have an increased risk for hemorrhagic strokes with aspirin use. A new RCT and meta-analysis suggest that the risk for hemorrhagic strokes in women is not statistically significantly increased. LIMITATIONS: New evidence on aspirin for the primary prevention of CVD is limited. The dose of aspirin used in the RCTs varied, which prevented the estimation of the most appropriate dose for primary prevention. Several of the RCTs were conducted within populations of health professionals, which potentially limits generalizability. CONCLUSION: Aspirin reduces the risk for myocardial infarction in men and strokes in women. Aspirin use increases the risk for serious bleeding events. PMID- 19293074 TI - Nursing home physician specialists: a response to the workforce crisis in long term care. AB - Marginalization of physicians in the nursing home threatens the overall care of increasingly frail nursing home residents who have medically complex illnesses. The authors propose that creating a nursing home medicine specialty, which recognizes the nursing home as a unique practice site, would go a long way toward remedying existing problems with care in skilled nursing facilities and would best serve the needs of the 1.6 million nursing home residents in the United States. Reviewing what is known about physician practice in nursing homes and hospitals, and taking a lead from the hospitalist movement, the specialty would be characterized in 3 dimensions: the degree of physicians' commitment, physicians' practice competencies, and the structure of the medical staff organization in which they practice. Challenges to the adoption of a nursing home specialist model include mainstream medicine's failure to recognize the nursing home as a legitimate medical practice, the need for the nursing home industry and policymakers to appreciate the links between physician practice and quality, and assurance of financial viability. Implications for quality of care, health policy, and research needs are discussed in this article. PMID- 19293075 TI - Aspirin for prevention and treatment of cardiovascular disease. PMID- 19293076 TI - Lost in translation. PMID- 19293077 TI - Just playing doctor. PMID- 19293078 TI - Questioning the accuracy of a recent review of osteoporosis medications. PMID- 19293079 TI - How much did biases in the study of chronic obstructive pulmonary disease medications and mortality affect the outcome? PMID- 19293080 TI - How much did biases in the study of chronic obstructive pulmonary disease medications and mortality affect the outcome? PMID- 19293081 TI - How much did biases in the study of chronic obstructive pulmonary disease medications and mortality affect the outcome? PMID- 19293082 TI - Are risk stratification tables the best way to evaluate model performance? PMID- 19293083 TI - The predictive value of the ELISpot-based interferon-gamma-release assay for tuberculosis disease. PMID- 19293084 TI - Chronic hepatitis after hepatitis E virus infection in a patient with non-Hodgkin lymphoma taking rituximab. PMID- 19293085 TI - Challenges in reporting meta-analyses of diagnostic accuracy studies. PMID- 19293086 TI - Incidence of asymptomatic patent foramen ovale according to age. PMID- 19293087 TI - More cases of drug-resistant tuberculosis complicate care but spur global research and innovation. PMID- 19293088 TI - Bayesian t tests for accepting and rejecting the null hypothesis. AB - Progress in science often comes from discovering invariances in relationships among variables; these invariances often correspond to null hypotheses. As is commonly known, it is not possible to state evidence for the null hypothesis in conventional significance testing. Here we highlight a Bayes factor alternative to the conventional t test that will allow researchers to express preference for either the null hypothesis or the alternative. The Bayes factor has a natural and straightforward interpretation, is based on reasonable assumptions, and has better properties than other methods of inference that have been advocated in the psychological literature. To facilitate use of the Bayes factor, we provide an easy-to-use, Web-based program that performs the necessary calculations. PMID- 19293089 TI - Reconceptualizing inhibition of return as habituation of the orienting response. AB - Inhibition of return (IOR) is an effect on spatial attention whereby reaction times to a target presented at a location where a stimulus had recently been presented are increased, as opposed to when a target is presented at a new location. Despite early reports that habituation is not responsible for the IOR effect, the human cognitive literature provides indirect evidence in favor of the possibility. In addition, recent neurophysiological studies provide direct support for the idea that habituation is at least a contributing source for the IOR effect. The present article describes how habituation may account for the IOR effect and explores some of the predictions that this hypothesis suggests. PMID- 19293090 TI - Super-recognizers: people with extraordinary face recognition ability. AB - We tested 4 people who claimed to have significantly better than ordinary face recognition ability. Exceptional ability was confirmed in each case. On two very different tests of face recognition, all 4 experimental subjects performed beyond the range of control subject performance. They also scored significantly better than average on a perceptual discrimination test with faces. This effect was larger with upright than with inverted faces, and the 4 subjects showed a larger "inversion effect" than did control subjects, who in turn showed a larger inversion effect than did developmental prosopagnosics. This result indicates an association between face recognition ability and the magnitude of the inversion effect. Overall, these "super-recognizers" are about as good at face recognition and perception as developmental prosopagnosics are bad. Our findings demonstrate the existence of people with exceptionally good face recognition ability and show that the range of face recognition and face perception ability is wider than has been previously acknowledged. PMID- 19293091 TI - Does response interference contribute to face composite effects? AB - Holistic processing of faces can be measured as a failure of selective attention to one face-half under instructions to ignore the other face-half in a naming or same/different matching task. But is interference from the irrelevant half due to response interference rather than to holistic processing? Here, participants learned to name two faces "Fred" and two "Bob." At test, composites were created from top and bottom halves of different learned faces or of a novel face, and composites were either aligned or misaligned. Naming was slower when the irrelevant half was from a different face as opposed to the same face, regardless of whether it was associated with the same name, a different name, or no name, suggesting holistic processing. Interference was eliminated when composite halves were misaligned. These results suggest that, unlike Stroop effects, composite effects are not due to response interference. PMID- 19293092 TI - Evidence for a contact-based explanation of the own-age bias in face recognition. AB - Previous research has shown that we recognize faces similar in age to ourselves better than older or younger faces (e.g., Wright & Stroud, 2002). This study investigated whether this own-age bias could be explained by the contact hypothesis used to account for the own-race bias (see Meissner & Brigham, 2001). If the own-age bias stems from increased exposure to people of our own age, it should be reduced or absent in those with higher exposure to other age groups. Participants were asked to remember facial photographs of 8- to 11- and 20- to 25 year-olds. Undergraduates were faster and more accurate at recognizing faces of their own age. However, trainee teachers showed no such own-age bias; they recognized the children's faces more quickly than own-age faces and with comparable accuracy. These results support a contact-based explanation of the own age bias. PMID- 19293093 TI - Race-based perceptual asymmetries underlying face processing in infancy. AB - Adults process other-race faces differently than own-race faces. For instance, a single other-race face in an array of own-race faces attracts Caucasians' attention, but a single own-race face among other-race faces does not. This perceptual asymmetry has been explained by the presence of an other-race feature in other-race faces and its absence in own-race faces; this difference is thought to underlie race-based differences in face processing. We examined the developmental origins of this mechanism in two groups of Caucasian 9-month-olds. Infants in the experimental group exhibited a preference for a pattern containing a single Asian face among seven Caucasian faces over a pattern containing a single Caucasian face among seven Asian faces. This preference was not driven by the majority of elements in the images, because a control group of infants failed to exhibit a preference between homogeneous patterns containing eight Caucasian versus eight Asian faces. The results demonstrate that an other-race face among own-race faces attracts infants' attention but not vice versa. This perceptual asymmetry suggests that the other-race feature is available to Caucasians by 9 months of age, thereby indicating that mechanisms of specialization in face processing originate early in life. PMID- 19293094 TI - Can beauty be ignored? Effects of facial attractiveness on covert attention. AB - Facial beauty has important social and biological implications. Research has shown that people tend to look longer at attractive than at unattractive faces. However, little is known about whether an attractive face presented outside foveal vision can capture attention. The effect of facial attractiveness on covert attention was investigated in a spatial cuing task. Participants were asked to judge the orientation of a cued target presented to the left or right visual field while ignoring a task-irrelevant face image flashed in the opposite field. The presentation of attractive faces significantly lengthened task performance. The results suggest that facial beauty automatically competes with an ongoing cognitive task for spatial attention. PMID- 19293095 TI - Nonautomatic emotion perception in a dual-task situation. AB - Are emotions perceived automatically? Two psychological refractory period experiments were conducted to ascertain whether emotion perception requires central attentional resources. Task 1 required an auditory discrimination (tone vs. noise), whereas Task 2 required a discrimination between happy and angry faces. The difficulty of Task 2 was manipulated by varying the degree of emotional expression. The stimulus onset asynchrony (SOA) between Task 1 and Task 2 was also varied. Experiment 1 revealed additive effects of SOA and Task 2 emotion-perception difficulty. Experiment 2 replicated the additive relationship with a stronger manipulation of emotion-perception difficulty. According to locus of-slack logic, our participants did not process emotional expressions while central resources were devoted to Task 1. We conclude that emotion perception is not fully automatic. PMID- 19293096 TI - Three-quarter views are subjectively good because object orientation is uncertain. AB - Because the objects that surround us are three-dimensional, their appearance and our visual perception of them change depending on an object's orientation relative to a viewpoint. One of the most remarkable effects of object orientation is that viewers prefer three-quarter views over others, such as front and back, but the exact source of this preference has not been firmly established. We show that object orientation perception of the three-quarter view is relatively imprecise and that this impreciseness is related to preference for this view. Human vision is largely insensitive to variations among different three-quarter views (e.g., 45 degrees vs. 50 degrees ); therefore, the three-quarter view is perceived as if it corresponds to a wide range of orientations. In other words, it functions as the typical representation of the object. PMID- 19293097 TI - Facilitated pointing to remembered objects in front: evidence for egocentric retrieval or for spatial priming? AB - Recent spatial memory theories propose that long-term spatial memories are retrieved egocentrically. One source of evidence comes from imagined perspective taking, in which participants learn an object layout, later imagine standing at one object and facing a second (orienting) object, and then point to a third (target) object from the imagined perspective. Pointing is faster for target objects in the anterior than in the posterior half of imaginal space. This "front facilitation" is consistent with asymmetric sensory and biomechanical body properties (favoring the anterior half of body space), supporting claims of egocentric retrieval. However, front facilitation might actually result from spatial priming: Proximity differences might cause orienting objects to prime target objects more in the anterior than in the posterior half of imagined space. Using a modified perspective-taking task that unconfounded front facilitation and spatial priming, two experiments identified separate influences of front facilitation and spatial priming when participants imagined perspectives within the surrounding environment or a remote environment. PMID- 19293098 TI - Orientation dependence of spatial memory acquired from auditory experience. AB - The present study investigated whether memory for a room-sized spatial layout learned through auditory localization of sounds exhibits orientation dependence similar to that observed for spatial memory acquired from stationary viewing of the environment. Participants learned spatial layouts by viewing objects or localizing sounds and then performed judgments of relative direction among remembered locations. The results showed that direction judgments following auditory learning were performed most accurately at a particular orientation in the same way as were those following visual learning, indicating that auditorily encoded spatial memory is orientation dependent. In combination with previous findings that spatial memories derived from haptic and proprioceptive experiences are also orientation dependent, the present finding suggests that orientation dependence is a general functional property of human spatial memory independent of learning modality. PMID- 19293099 TI - Facilitation of learning spatial relations among locations by visual cues: implications for theoretical accounts of spatial learning. AB - Human participants searched in a real environment or interactive 3-D virtual environment open field for four hidden goal locations arranged in a 2 x 2 square configuration in a 5 x 5 matrix of raised bins. The participants were randomly assigned to one of two groups: cues + pattern or pattern only. The participants experienced a training phase, followed by a testing phase. Visual cues specified the goal locations during training only for the cues + pattern group. Both groups were then tested in the absence of visual cues. The results in both environments indicated that the participants learned the spatial relations among goal locations. However, visual cues during training facilitated learning of the spatial relations among goal locations: In both environments, the participants trained with the visual cues made fewer errors during testing than did those trained only with the pattern. The results suggest that learning based on the spatial relations among locations may not be susceptible to cue competition effects and have implications for standard associative and dual-system accounts of spatial learning. PMID- 19293100 TI - Electrophysiological correlates of high-level perception during spatial navigation. AB - We studied the electrophysiological basis of object recognition by recording scalp electroencephalograms while participants played a virtual-reality taxi driver game. Participants searched for passengers and stores during virtual navigation in simulated towns. We compared oscillatory brain activity in response to store views that were targets or nontargets (during store search) or neutral (during passenger search). Even though store category was solely defined by task context (rather than by sensory cues), frontal electrophysiological activity in the low frequency bands (primarily in the [4-8 Hz] band) reliably distinguished between the target, nontarget, and neutral store views. These results implicate low-frequency oscillatory brain activity in frontal regions as an important variable in the study of the cognitive processes involved in object recognition, categorization, and other forms of high-level perception. PMID- 19293101 TI - Spatial negative priming in bilingualism. AB - Balanced bilinguals have been shown to have an enhanced ability to inhibit distracting information. In this study, we investigated the hypothesis that the bilinguals' efficiency in inhibitory control can be advantageous in some conditions, but disadvantageous in others-for example, negative priming conditions, in which previously irrelevant information becomes relevant. Data collected in a target-stimulus locating task from 29 early bilingual adults and 29 age-matched monolinguals showed that the bilinguals' greater inhibition of irrelevant spatial information (i.e., the position of a distractor stimulus) resulted in a smaller effect of the distractor presence (i.e., a smaller difference in error rates in trials with and without distractors) and a larger negative priming effect (i.e., a larger difference between the error rates shown in trials wherein the target position corresponded to the position of the previous-trial distractor and trials wherein the target was presented in a previously vacant position). These findings support the hypothesis of specific nonlinguistic cognitive effects of bilingualism on inhibitory control functions, which are not necessarily reflected in cognitive advantages. PMID- 19293102 TI - Reading habits for both words and numbers contribute to the SNARC effect. AB - This study compared the spatial representation of numbers in three groups of adults: Canadians, who read both English words and Arabic numbers from left to right; Palestinians, who read Arabic words and Arabic-Indic numbers from right to left; and Israelis, who read Hebrew words from right to left but Arabic numbers from left to right. Canadians associated small numbers with left and large numbers with right space (the SNARC effect), Palestinians showed the reverse association, and Israelis had no reliable spatial association for numbers. These results suggest that reading habits for both words and numbers contribute to the spatial representation of numbers. PMID- 19293103 TI - Integers do not automatically activate their quantity representation. AB - Researchers have generally come to the conclusion that integers automatically activate the quantity they symbolize and that this quantity dominates responding. I conducted a strong test of this hypothesis with two numerical same/different experiments. On each trial, I presented the participant an integer between 1 and 9 and asked him or her to identify whether that symbol was a 5. If quantity information dominates responding, participants' reaction times (RTs) should be a function of the numerical distance between the target and the distractor. If quantity information is not activated, the integer is merely a shape, and participants' RTs should be a function of the physical similarity of the target and the distractor. The data from Experiments 1 and 2 demonstrate that quantity information exerts no control and that physical similarity is the primary controlling factor. These findings demonstrate that integers maintain a level of independence from their quantity representations. PMID- 19293104 TI - How many exemplars are used? Explorations with the Rex Leopold I model. AB - The goal of this research is to test the hypothesis that a category is not necessarily represented by all observed exemplars, but by a reduced subset of these exemplars. To test this hypothesis, we made use of a study reported by Nosofsky, Clark, and Shin (1989), and replicated their Experiment 1 in order to gather individual-participant data. Both a full exemplar model and a reduced exemplar model were fit to the data. In general, the fits of the reduced exemplar model were superior to those of the full exemplar model. The results suggest that only a subset of exemplars may be sufficient for category representation. PMID- 19293105 TI - What is pressure? Evidence for social pressure as a type of regulatory focus. AB - Previous research (Markman, Maddox, & Worthy, 2006) suggests that pressure leads to choking when one is learning to classify items on the basis of an explicit rule, but it leads to excelling when one is learning to classify items on the basis of an implicit strategy. In this article, we relate social pressure to regulatory focus theory. We propose that the effects of pressure on performance arise because pressure induces a prevention focus that interacts with the more local reward structure of the task. To test this hypothesis, we repeated previous research, but using a losses reward structure, so that participants under pressure were in a regulatory fit. We also successfully replicated previous results by using a gains reward structure. In contrast with participants who attempted to maximize gains on each trial, participants who attempted to minimize losses choked on the implicit-learning task but excelled on the explicit-learning task. The results suggest a three-way interaction between pressure level, task type, and reward structure. PMID- 19293106 TI - Adjustments to recent and frequent conflict reflect two distinguishable mechanisms. AB - In conflict paradigms such as the Eriksen flanker task, interference has been found to be reduced under conditions of recent and/or frequent cognitive conflict. Using a modified flanker task, we investigated the interplay of conflict recency and conflict frequency by comparing the interference reductions following conflict trials under conditions in which conflict was either frequent or infrequent overall, while controlling for stimulus and response feature repetitions to rule out nonattentional accounts. The reduction of flanker interference after a conflict trial was attenuated when overall conflict was frequent. This result is consistent with models assuming that processing adjustments occur gradually in response to conflict strength, such as the connectionist model of Botvinick, Braver, Barch, Carter, and Cohen (2001), in which both recency and frequency modulations are brought about by the same mechanism. By decomposing response times into initiation times and movement times, we revealed that frequent conflict delayed response initiation but sped up movement. Moreover, whereas frequent conflict reduced interference in both components, interference reduction after individual conflict trials was confined to movement times. Taken together, these results suggest that different mechanisms underlie the two kinds of modulation. PMID- 19293107 TI - Sequential adjustments before and after partial errors. AB - In choice reaction time tasks, subjects speed up before making an error, but slow down afterward to prevent the occurrence of a new error. In some trials, the correct response is preceded by an incorrect electromyographic (EMG) activation too small to reach the response threshold. In this article, we show that these incorrect EMG activations give rise to the same sequential effects as overt errors: Before a trial containing an incorrect EMG activation, subjects speed up, whereas after that trial, they slow down. These activations reflect errors that have been detected, inhibited, and corrected in time. As such, they index the involvement of online executive control. PMID- 19293108 TI - Orthographic influences in spoken word recognition: the consistency effect in semantic and gender categorization tasks. AB - According to current models, spoken word recognition is driven by the phonological properties of the speech signal. However, several studies have suggested that orthographic information also influences recognition in adult listeners. In particular, it has been repeatedly shown that, in the lexical decision task, words that include rimes with inconsistent spellings (e.g., /-ip/ spelled -eap or -eep) are disadvantaged, as compared with words with consistent rime spelling. In the present study, we explored whether the orthographic consistency effect extends to tasks requiring people to process words beyond simple lexical access. Two different tasks were used: semantic and gender categorization. Both tasks produced reliable consistency effects. The data are discussed as suggesting that orthographic codes are activated during word recognition, or that the organization of phonological representations of words is affected by orthography during literacy acquisition. PMID- 19293109 TI - Neighborhood effects in spelling in adults. AB - The participants were asked to spell aloud words for which there were either many orthographically similar words (a dense neighborhood) or few orthographically similar words (a sparse neighborhood). Words with a dense neighborhood were spelled faster and more accurately than were words with a sparse neighborhood. This finding is consistent with the hypothesis of Rapp, Epstein, and Tainturier (2002), that the cognitive spelling system has an interactive architecture incorporating feedback between individual graphemes and orthographic lexeme representations. PMID- 19293110 TI - Making psycholinguistics musical: self-paced reading time evidence for shared processing of linguistic and musical syntax. AB - Linguistic processing, especially syntactic processing, is often considered a hallmark of human cognition; thus, the domain specificity or domain generality of syntactic processing has attracted considerable debate. The present experiments address this issue by simultaneously manipulating syntactic processing demands in language and music. Participants performed self-paced reading of garden path sentences, in which structurally unexpected words cause temporary syntactic processing difficulty. A musical chord accompanied each sentence segment, with the resulting sequence forming a coherent chord progression. When structurally unexpected words were paired with harmonically unexpected chords, participants showed substantially enhanced garden path effects. No such interaction was observed when the critical words violated semantic expectancy or when the critical chords violated timbral expectancy. These results support a prediction of the shared syntactic integration resource hypothesis (Patel, 2003), which suggests that music and language draw on a common pool of limited processing resources for integrating incoming elements into syntactic structures. Notations of the stimuli from this study may be downloaded from pbr.psychonomic journals.org/content/supplemental. PMID- 19293111 TI - Perceptual learning in auditory temporal discrimination: no evidence for a cross modal transfer to the visual modality. AB - Perceptual learning was used to study potential transfer effects in a duration discrimination task. Subjects were trained to discriminate between two empty temporal intervals marked with auditory beeps, using a two-alternative forced choice paradigm. The major goal was to examine whether perceptual learning would generalize to empty intervals that have the same duration but are marked by visual flashes. The experiment also included longer intervals marked with auditory beeps and filled auditory intervals of the same duration as the trained interval, in order to examine whether perceptual learning would generalize to these conditions within the same sensory modality. In contrast to previous findings showing a transfer from the haptic to the auditory modality, the present results do not indicate a transfer from the auditory to the visual modality; but they do show transfers within the auditory modality. PMID- 19293112 TI - Expert image analysts show enhanced visual processing in change detection. AB - Expertise facilitates change detection performance, but the neural underpinnings of these benefits are unknown. Expert image analysts showed larger change-related ERP effects between about 100-200 msec after stimulus onset than did novices, which correlated with both accuracy and years of analysis experience. These results demonstrate that years of visual experience can induce fundamental changes in early visual processing which are related to change detection abilities. PMID- 19293113 TI - The effects of individual differences and task difficulty on inattentional blindness. AB - Most studies of inattentional blindness-the failure to notice an unexpected object when attention is focused elsewhere-have focused on one critical trial. For that trial, noticing the unexpected object might be a result of random variability, so that any given individual would be equally likely to notice the unexpected object. On the other hand, individual differences in the ability to perform the primary task might make noticing more likely for some individuals than for others. Increasing the difficulty of the primary task has been shown to decrease noticing rates for both brief static displays (Cartwright-Finch & Lavie, 2007) and dynamic monitoring tasks (Simons & Chabris, 1999). However, those studies did not explore whether individual differences in noticing arise from differences in the ability to perform the primary task. For our Experiment 1, we used a staircase procedure to equate primary task performance across individuals in a dynamic inattentional blindness task and found that the demands of the primary task affected noticing rates when individual differences in accuracy were minimized. In Experiment 2, we found that individual differences in primary task performance did not predict noticing of an unexpected object. Together, these findings suggest that although the demands of the primary task do affect inattentional blindness rates, individual differences in the ability to meet those demands do not. PMID- 19293114 TI - Perceptual load modulates attentional capture by abrupt onsets. AB - The abrupt appearance of a new object captures attention, even when the object is task irrelevant. These findings suggest that abrupt onsets capture attention in a stimulus-driven manner and are not susceptible to top-down influences on attentional control. However, previous studies examining the ability of abrupt onsets to capture attention have used search displays that lacked significant complexity. Because attention is a limited capacity mechanism, it is possible that increasing the complexity, or perceptual load, of the search arrays may modulate capture by abrupt onsets. We used a flanker task to examine the effect of perceptual load on attentional capture by abruptly appearing objects. Subjects searched for a target letter through low-load (set size = 1) and high-load (set size = 6) displays. On each trial, irrelevant flankers also appeared, one as an onset and the other as an offset. Onset flankers affected search in low-load but not high-load displays. This modulation of attentional capture was not caused by generalized slowing when subjects searched through high-load displays; search for a single perceptually degraded target slowed response times but did not affect attentional capture. These findings demonstrate that attentional capture by an abrupt onset is attenuated when people search through high-load scenes. PMID- 19293115 TI - Attentional enhancement during multiple-object tracking. AB - What is the role of attention in multiple-object tracking? Does attention enhance target representations, suppress distractor representations, or both? It is difficult to ask this question in a purely behavioral paradigm without altering the very attentional allocation one is trying to measure. In the present study, we used event-related potentials to examine the early visual evoked responses to task-irrelevant probes without requiring an additional detection task. Subjects tracked two targets among four moving distractors and four stationary distractors. Brief probes were flashed on targets, moving distractors, stationary distractors, or empty space. We obtained a significant enhancement of the visually evoked P1 and N1 components (approximately 100-150 msec) for probes on targets, relative to distractors. Furthermore, good trackers showed larger differences between target and distractor probes than did poor trackers. These results provide evidence of early attentional enhancement of tracked target items and also provide a novel approach to measuring attentional allocation during tracking. PMID- 19293116 TI - Learning to ignore: acquisition of sustained attentional suppression. AB - We examined whether the selection mechanisms committed to the suppression of ignored stimuli can be modified by experience to produce a sustained, rather than transient, change in behavior. Subjects repeatedly ignored the shape of stimuli, while attending to their color. On subsequent attention to shape, there was a robust and sustained decrement in performance that was selective to when shape was ignored across multiple-color-target contexts, relative to a single-color target context. Thus, amount of time ignored was not sufficient to induce a sustained performance decrement. Moreover, in this group, individual differences in initial color target selection were associated with the subsequent performance decrement when attending to previously ignored stimuli. Accompanying this sustained decrement in performance was a transfer in the locus of suppression from an exemplar (e.g., a circle) to a feature (i.e., shape) level of representation. These data suggest that learning can influence attentional selection by sustained attentional suppression of ignored stimuli. PMID- 19293117 TI - P(rep) misestimates the probability of replication. AB - The probability of "replication," P(rep), has been proposed as a means of identifying replicable and reliable effects in the psychological sciences. We conduct a basic test of P(rep) that reveals that it misestimates the true probability of replication, especially for small effects. We show how these general problems with P(rep) play out in practice, when it is applied to predict the replicability of observed effects over a series of experiments. Our results show that, over any plausible series of experiments, the true probabilities of replication will be very different from those predicted by P(rep). We discuss some basic problems in the formulation of P(rep) that are responsible for its poor performance, and conclude that P(rep) is not a useful statistic for psychological science. PMID- 19293118 TI - Leadership's influence on job satisfaction. AB - PURPOSE: To assess the leadership styles of frontline medical imaging supervisors and examine the relationship between leadership behaviors and motivational factors that influence job satisfaction of medical imaging staff in acute care facilities. METHODS: Three hundred fifty nine staff technologists completed a survey on their supervisors' leadership behaviors and the intrinsic and extrinsic motivating factors that influence job satisfaction and organizational commitment. RESULTS: This study indicates that there are strong, positive relationships between supervisors' transformational and contingent reward behaviors and intrinsic esteem motivators that influence staff members' job satisfaction. The behaviors with the strongest relationship to intrinsic esteem motivators were individualized consideration and contingent rewards. CONCLUSION: The results of this study provide insight into what employees need from their leader and the organization that employs them. This information can be used to help develop strategies to meet those needs through work redesign and leadership behavior development. PMID- 19293119 TI - Assessing radiologist assistant supervision levels in Florida. AB - BACKGROUND: As radiologist assistants (RAs) become more prevalent in the radiology work force, it is important to re-evaluate how they practice and are supervised. PURPOSE: To analyze the impact of RA supervision levels on clinical practice. METHODS: A survey was conducted to evaluate whether a random sample of radiologists in Florida would recommend changes to the current levels of supervision required for advanced-practice technologists. The survey form solicited radiologists' opinions concerning practice activities an advanced practice radiographer would perform and the appropriate supervision levels for each of those activities. RESULTS: Reduced levels of supervision were recommended by the respondents for 62.5% of the changes to procedures overall, although the responses varied considerably across the procedures. Recommendations for change were more likely when the radiologist had experience working with a physician extender and for supervision of certain types of procedures. DISCUSSION: The study revealed that as more experience is gained working with RAs, radiologists increasingly are supportive of lowering supervision levels. PMID- 19293120 TI - Genetics and genomic medicine. AB - Genetics is playing an increasingly important role in the diagnosis, monitoring and treatment of diseases, and the expansion of genetics into health care has generated the field of genomic medicine. Health care delivery is shifting away from general diagnostic evaluation toward a generation of therapeutics based on a patient's genetic makeup. Meanwhile, the scientific community debates how best to incorporate genetics and genomic medicine into practice. While obstacles remain, the ultimate goal is to use information generated from the study of human genetics to improve disease treatment, cure and prevention. As the use of genetics in medical diagnosis and treatment increases, health care workers will require an understanding of genetics and genomic medicine. PMID- 19293121 TI - Imaging diabetes. AB - The American Diabetes Association recently released a report on the economic costs of diabetes in the United States. Authors of the report concluded that in 2007 the national economic burden of diabetes exceeded $174 billion. However, even such sobering statistics cannot quantify the emotional toll diabetes exacts on patients, their friends and families. With the annual prevalence of diabetes continuing to exceed all predictions, imaging is increasingly called upon to assist in disease management, exploration of mechanisms underlying the disease process and evaluation of the effectiveness of novel treatments. PMID- 19293122 TI - Chiari malformations. AB - Chiari malformations can be serious conditions and their diagnosis often confounds clinicians. When patients present with Chiari malformations, they may have no symptoms or a range of symptoms, many of which can be confused with other neurological conditions. Imaging, and particularly magnetic resonance, plays an important role in diagnosing and treating Chiari malformations. PMID- 19293123 TI - Student analysis of clinical education. PMID- 19293124 TI - Pelvic thumbprint? PMID- 19293126 TI - Imaging retained surgical instruments. PMID- 19293127 TI - Information literacy. PMID- 19293128 TI - Sonography in Malawi. PMID- 19293129 TI - Sinus series. PMID- 19293130 TI - Switching from donepezil tablets to rivastigmine transdermal patch in Alzheimer's disease. AB - OBJECTIVE: Evaluate safety and tolerability of switching from donepezil to rivastigmine transdermal patch in patients with mild to moderate Alzheimer's disease. METHODS: Prospective, parallel-group, open-label study to evaluate immediate or delayed switch from 5-10 mg/day donepezil to 4.6 mg/24 h rivastigmine following a 4-week treatment period. RESULTS: Rates of discontinuation due to any reason or adverse events were similar between groups. Incidences of gastrointestinal adverse events were 3.8% in the immediate and 0.8% in the delayed switch group. No patients discontinued secondary to nausea and vomiting. Discontinuations due to application site reactions were low (2.3%). Asymptomatic bradycardia was more common following the immediate switch (2.3% vs 0%); however, these patients had coexisting cardiac comorbidities. CONCLUSION: Both switch strategies were safe and well tolerated. The majority of patients may be able to switch directly to rivastigmine patches without a withdrawal period. Appropriate clinical judgment should be used for patients with existing bradycardia or receiving beta blockers. PMID- 19293131 TI - Increased expression of ac-FoxO1 protein in prelabor fetal membranes overlying the cervix: possible role in human fetal membrane rupture. AB - Forkhead box O proteins have critical roles in a number of cellular processes, including apoptosis. Acetylation and phosphorylation of forkhead box O proteins are posttranslational modifications that attenuate their transcriptional activity. As supracervical fetal membranes are characterized by increased cell death, the aim of this study was to compare the expression of forkhead box O1, acetylated-forkhead box O1, and Ser-256 phosphorylated forkhead box O1 at supracervical and distal site fetal membrane. Fetal membranes overlying the cervix were identified in situ in women undergoing term elective Caesarean section. Immunohistochemistry (n = 7) was used to analyze the protein expression of forkhead box O1, acetylated-forkhead box O1, and Ser-256 phosphorylated forkhead box O1. There was no difference in forkhead box O1 and Ser-256 phosphorylated forkhead box O1 protein expression between the 2 sites. However, when compared with distal site, the intensity and extent of staining of acetylated-forkhead box O1 were greater in amnion and chorion obtained from the supracervical site. In summary, supracervical fetal membranes are characterized by increased acetylated-forkhead box O1 protein expression. Although the precise role and contribution of acetylated-forkhead box O1 in the process of human fetal membrane rupture are unknown, it has been implicated in apoptosis and/or cell cycle regulation. PMID- 19293132 TI - Review: cellular metabolism: contribution to postoperative adhesion development. AB - Postoperative adhesions are a significant source of morbidity, including contributions to pelvic pain, bowel obstruction, and infertility. While the mechanisms of postoperative adhesion development are complex and incompletely understood, hypoxia appears to trigger a cascade of intracellular responses involving hypoxia-inducible factors, lactate, reactive oxygen species, reactive nitrogen species, and insulin-like growth factors that results in manifestation of the adhesion phenotype. Thus, substantial evidence exists to implicate the direct role of cellular metabolism in wound repair and adhesion development. PMID- 19293133 TI - Effect of periconceptional undernutrition in sheep on late gestation expression of mRNA and protein from genes involved in fetal adrenal steroidogenesis and placental prostaglandin production. AB - In sheep, maternal periconceptional undernutrition precociously activates fetal hypothalamic-pituitary-adrenal function in a high proportion of animals, resulting in preterm birth. We investigated whether the effects of periconceptional undernutrition were mediated through genes encoding enzymes involved in adrenal steroidogenesis and in placental production and metabolism of steroids and prostaglandins. Singleton-bearing ewes were fed ad libitum (control, N) throughout gestation or undernourished from 60 days before until 30 days after mating. Fetal adrenal and placentome tissues were collected on gestational day 131. Fetal adrenal P450C17 protein expression was significantly increased with undernutrition. Placental prostaglandin endoperoxide synthase-2 expression was not different between groups. In undernourished pregnancies, there were significant positive correlations between fetal plasma adrenocorticotrophic hormone and cortisol concentrations and between fetal plasma adrenocorticotrophic hormone concentrations and placental prostaglandin endoperoxide synthase-2 messenger RNA (mRNA) and protein. These studies suggest that periconceptional undernutrition affects fetal adrenal P450C17, consistent with an elevation in plasma cortisol, and this occurs prior to activation of placental prostaglandin endoperoxide synthase-2 expression at gestational day 131. PMID- 19293134 TI - Renal biopsy in liver transplant recipients. AB - BACKGROUND: Renal impairment post-liver transplant (LT) is often attributed to calcineurin inhibitors (CNIs). A renal biopsy can be a useful tool but may be complicated in LT recipients. We aimed to determine the clinical scenarios that prompted a decision to perform a renal biopsy in this patient population, to assess histological findings and evaluate patient management and survival and renal outcome. METHODS: Information on clinical variables and renal histology was extracted from single-centre prospectively compiled databases from 1996 onwards. RESULTS: Over 2100 adults received an LT in the time period studied, and 54 of these (35 males and 19 females) were referred for renal review. Of these, 43% underwent a renal biopsy. They had a higher creatinine (P = 0.02), a greater deterioration in creatinine over the year prior to review and were more likely to be nephrotic (both P < 0.01). Histological findings included hypertensive changes (44%), CNI nephrotoxicity (48%), IgA nephropathy (9%), membranoproliferative glomerulonephritis (17%), acute tubular necrosis (4%), crescentic glomerulonephritis (4%) and diabetic nephropathy (9%). Major bleeding complications occurred in 17%. Treatment changed in the majority but, it was not significantly different in the two groups. Although initial renal function was worse in the biopsied group, final patient and renal survival did not differ between the two groups. CONCLUSION: A renal biopsy is a valuable tool in those with renal insufficiency and/or proteinuria and haematuria but the benefits must be weighed against the relatively high complication rate in LT recipients. PMID- 19293135 TI - A critical look at acute decompensated heart failure from a nephrologist's perspective. PMID- 19293136 TI - Nephropathy in Fabry disease: the importance of early diagnosis and testing in high-risk populations. PMID- 19293137 TI - Sudden death and associated factors in a historical cohort of chronic haemodialysis patients. AB - BACKGROUND: In haemodialysis patients, deaths due to cardiovascular causes constitute a large proportion of total mortality and sudden cardiac deaths account for approximately 22% of all deaths. The aim of this study was to evaluate the incidence of sudden cardiac death and associated risk factors in a cohort of haemodialysis patients. METHODS AND RESULTS: The 3-year cumulative incidence of death in a cohort of 476 patients on chronic haemodialysis treatment was 34.3% (SE 2.3). Sudden death had a 6.9% (SE 1.2) cumulative incidence, with 32 events representing 19.2% of all deaths, while cardiovascular not sudden death and noncardiovascular death accounted for a 3-year cumulative incidence of 7.3% (SE 1.2) and 20.1% (SE 1.9), respectively. According to Cox multivariate analysis, significant risk factors for sudden death were the presence of atrial fibrillation, diabetes mellitus, predialytic hyperkalaemia, haemodialysis mode and C-reactive protein level, which were associated with a 2.9 (CI(95%) 1.3-6.4), 3.0 (CI(95%) 1.3-7.2), 2.7 (CI(95%) 1.3-5.8), 4.5 (CI(95%) 1.3-15.5) and 3.3 (CI(95%) 1.2-8.8)-fold increase in the risk of sudden death, respectively. Sudden death was significantly more frequent during the first 24 h of the first short interdialytic interval and during the last 24 h of the long interval, i.e. immediately before and immediately after the first weekly haemodialysis session (P = 0.02). CONCLUSIONS: Our data show that the incidence of sudden death in haemodialysis patients is high and that atrial fibrillation, diabetes, hyperkalaemia, haemodialysis mode and C-reactive protein play an important role in developing fatal arrhythmia. Further studies will be necessary to define which interventions could be helpful in reducing this cause of mortality. PMID- 19293138 TI - The multi-AT-hook chromosomal protein of Drosophila melanogaster, D1, is dispensable for viability. AB - The D1 protein is a high mobility group A (HMGA)-like nonhistone chromosomal protein with primary localization to certain AT-rich satellite DNA sequences within heterochromatin. The binding of D1 to euchromatic sequences is less studied and the functional significance of its chromosomal associations is unclear. By taking advantage of existing P-insertion alleles of the D1 gene, I generated D1 null mutations to investigate the phenotypic effect of loss of the D1 gene. In contrast to a previous report, I determined that the D1 gene is not essential for viability of Drosophila melanogaster, and moreover, that loss of D1 has no obvious phenotypic effects. My tests for an effect of D1 mutations on PEV revealed that it is not a suppressor of variegation, as concluded by other investigators. In fact, the consequence of loss of D1 on one of six variegating rearrangements tested, T(2;3)Sb(V), was dominant enhancement of PEV, suggesting a role for the protein in euchromatic chromatin structure and/or transcription. A study of D1 protein sequence conservation highlighted features shared with mammalian HMGA proteins, which function as architectural transcription factors. PMID- 19293139 TI - A proximal centriole-like structure is present in Drosophila spermatids and can serve as a model to study centriole duplication. AB - Most animals have two centrioles in spermatids (the distal and proximal centrioles), but insect spermatids seem to contain only one centriole (Fuller 1993), which functionally resembles the distal centriole. Using fluorescent centriolar markers, we identified a structure near the fly distal centriole that is reminiscent of a proximal centriole (i.e., proximal centriole-like, or PCL). We show that the PCL exhibits several features of daughter centrioles. First, a single PCL forms near the proximal segment of the older centriole. Second, the centriolar proteins SAS-6, Ana1, and Bld10p/Cep135 are in the PCL. Third, PCL formation depends on SAK/PLK4 and SAS-6. Using a genetic screen for PCL defect, we identified a mutation in the gene encoding the conserved centriolar protein POC1, which is part of the daughter centriole initiation site (Kilburn et al. 2007) in Tetrahymena. We conclude that the PCL resembles an early intermediate structure of a forming centriole, which may explain why no typical centriolar structure is observed under electron microscopy. We propose that, during the evolution of insects, the proximal centriole was simplified by eliminating the later steps in centriole assembly. The PCL may provide a unique model to study early steps of centriole formation. PMID- 19293140 TI - Predicting quantitative traits with regression models for dense molecular markers and pedigree. AB - The availability of genomewide dense markers brings opportunities and challenges to breeding programs. An important question concerns the ways in which dense markers and pedigrees, together with phenotypic records, should be used to arrive at predictions of genetic values for complex traits. If a large number of markers are included in a regression model, marker-specific shrinkage of regression coefficients may be needed. For this reason, the Bayesian least absolute shrinkage and selection operator (LASSO) (BL) appears to be an interesting approach for fitting marker effects in a regression model. This article adapts the BL to arrive at a regression model where markers, pedigrees, and covariates other than markers are considered jointly. Connections between BL and other marker-based regression models are discussed, and the sensitivity of BL with respect to the choice of prior distributions assigned to key parameters is evaluated using simulation. The proposed model was fitted to two data sets from wheat and mouse populations, and evaluated using cross-validation methods. Results indicate that inclusion of markers in the regression further improved the predictive ability of models. An R program that implements the proposed model is freely available. PMID- 19293141 TI - Resistance Gene Replacement in the mosquito Culex pipiens: fitness estimation from long-term cline series. AB - How adaptation appears and is later refined by natural selection has been the object of intense theoretical work. However, the testing of these theories is limited by our ability to estimate the strength of natural selection in nature. Using a long-term cline series, we estimate the selection coefficients acting on different alleles at the same locus to analyze the allele replacement observed in the insecticide resistance gene Ester in the mosquito Culex pipiens in the Montpellier area, southern France. Our method allows us to accurately account for the resistance allele replacement observed in this area since 1986. A first resistance allele appeared early, which was replaced by a second resistance allele providing the same advantage but at a lower cost, itself being replaced by a third resistance allele with both higher advantage and cost. It shows that amelioration of the adaptation (here resistance to insecticide) through allele replacement was successively achieved by selection of first a generalist allele (i.e., with a low fitness variance across environments) and later a specialist allele (i.e., with a large fitness variance across environments). More generally, we discuss how precise estimates of the strength of selection obtained from field data help us understand the process of amelioration of adaptation. PMID- 19293142 TI - Estimation of allele frequencies from high-coverage genome-sequencing projects. AB - A new generation of high-throughput sequencing strategies will soon lead to the acquisition of high-coverage genomic profiles of hundreds to thousands of individuals within species, generating unprecedented levels of information on the frequencies of nucleotides segregating at individual sites. However, because these new technologies are error prone and yield uneven coverage of alleles in diploid individuals, they also introduce the need for novel methods for analyzing the raw read data. A maximum-likelihood method for the estimation of allele frequencies is developed, eliminating both the need to arbitrarily discard individuals with low coverage and the requirement for an extrinsic measure of the sequence error rate. The resultant estimates are nearly unbiased with asymptotically minimal sampling variance, thereby defining the limits to our ability to estimate population-genetic parameters and providing a logical basis for the optimal design of population-genomic surveys. PMID- 19293143 TI - Analysis of the sequence and phenotype of Drosophila Sex combs reduced alleles reveals potential functions of conserved protein motifs of the Sex combs reduced protein. AB - The Drosophila Hox gene, Sex combs reduced (Scr), is required for patterning the larval and adult, labial and prothoracic segments. Fifteen Scr alleles were sequenced and the phenotypes analyzed in detail. Six null alleles were nonsense mutations (Scr(2), Scr(4), Scr(11), Scr(13), Scr(13A), and Scr(16)) and one was an intragenic deletion (Scr(17)). Five hypomorphic alleles were missense mutations (Scr(1), Scr(3), Scr(5), Scr(6), and Scr(8)) and one was a small protein deletion (Scr(15)). Protein sequence changes were found in four of the five highly conserved domains of SCR: the DYTQL motif (Scr(15)), YPWM motif (Scr(3)), Homeodomain (Scr(1)), and C-terminal domain (CTD) (Scr(6)), indicating importance for SCR function. Analysis of the pleiotropy of viable Scr alleles for the formation of pseudotracheae suggests that the DYTQL motif and the CTD mediate a genetic interaction with proboscipedia. One allele Scr(14), a missense allele in the conserved octapeptide, was an antimorphic allele that exhibited three interesting genetic properties. First, Scr(14)/Df had the same phenotype as Scr(+)/Df. Second, the ability of the Scr(14) allele to interact intragenetically with Scr alleles mapped to the first 82 amino acids of SCR, which contains the octapeptide motif. Third, Scr(6), which has two missense changes in the CTD, did not interact genetically with Scr(14). PMID- 19293144 TI - Validation of the Bayesian Alcoholism Test compared to single biomarkers in detecting harmful drinking. AB - AIMS: Conventional tests for alcohol dependence often fail to detect hazardous and harmful alcohol use (HHAU) accurately. We previously validated the Bayesian Alcoholism Test (BAT) for the detection of HHAU among males. This uses 15 biochemical and clinical variables, including questionnaire data to calculate the probability of harmful (>80 g alcohol/day), hazardous (40-80 g/day) and 'moderate' (<40 g/day) drinking. Here we investigate the BAT's diagnostic performance when more limited clinical data are available. METHODS: The WHO/ISBRA Collaborative Project recruited subjects from the general community and alcohol dependence treatment services. We analysed data from male drinkers: 318 alcohol dependent, 220 heavy and 712 moderate drinkers. Drinking was assessed using the Alcohol-Use Disorders and Associated Disabilities Interview Schedule. Eight of 15 markers used in the original BAT could be extracted from the WHO/ISBRA dataset. RESULTS: Comparing harmful to moderate drinkers, the area under the ROC curve for BAT (0.90) was significantly higher than that for CDT (0.82), GGT (0.77) and AST (0.76). Comparing hazardous to moderate drinkers, the area under the ROC curve for BAT (0.78) was significantly higher than that for AST (0.65) but not significantly higher than that for CDT (0.71) and GGT (0.70). For all 1250 subjects, the amount consumed correlated significantly better with BAT (0.65) than with CDT (0.52), GGT (0.44) or AST (0.40) alone. CONCLUSIONS: The BAT is more accurate than commonly used single biological markers in detecting harmful alcohol use, even when only half the input requirements are available. Computerized record keeping increases the practicality of use of algorithms in the detection of harmful drinking. PMID- 19293146 TI - Cerebral blood flow regulation by nitric oxide: recent advances. AB - Nitric oxide (NO) is undoubtedly quite an important intercellular messenger in cerebral and peripheral hemodynamics. This molecule, formed by constitutive isomers of NO synthase, endothelial nitric-oxide synthase, and neuronal nitric oxide synthase, plays pivotal roles in the regulation of cerebral blood flow and cell viability and in the protection of nerve cells or fibers against pathogenic factors associated with cerebral ischemia, trauma, and hemorrhage. Cerebral blood flow is increased and cerebral vascular resistance is decreased by NO derived from endothelial cells, autonomic nitrergic nerves, or brain neurons under resting and stimulated conditions. Somatosensory stimulation also evokes cerebral vasodilatation mediated by neurogenic NO. Oxygen and carbon dioxide alter cerebral blood flow and vascular tone mainly via constitutively formed NO. Endothelial dysfunction impairs cerebral hemodynamics by reducing the bioavailability of NO and increasing the production of reactive oxygen species (ROS). The NO-ROS interaction is an important issue in discussing blood flow and cell viability in the brain. Recent studies on brain circulation provide quite useful information concerning the physiological roles of NO produced by constitutive isoforms of nitric-oxide synthase and how NO may promote cerebral pathogenesis under certain conditions, including cerebral ischemia/stroke, cerebral vasospasm after subarachnoid hemorrhage, and brain injury. This information would contribute to better understanding of cerebral hemodynamic regulation and its dysfunction and to development of novel therapeutic measures to treat diseases of the central nervous system. PMID- 19293147 TI - Autobiographical memory in detoxified dependent drinkers. PMID- 19293148 TI - Thrombocytopenia in early alcohol withdrawal is associated with development of delirium tremens or seizures. AB - AIMS: In several studies, possible risk factors/predictors for severe alcohol withdrawal syndrome (AWS), i.e. delirium tremens (DT) and/or seizures, have been investigated. We have recently observed that low blood platelet count could be such a risk factor/predictor. We therefore investigated whether such an association could be found using a large number of alcohol-dependent individuals (n = 334). METHODS: This study is a retrospectively conducted cohort study based on data from female and male patients (>20 years of age), consecutively admitted to an alcohol treatment unit. The individuals had to fulfil the discharge diagnoses alcohol dependence and alcohol withdrawal syndrome according to DSM-IV. RESULTS: During the treatment period, 3% of the patients developed DT, 2% seizures and none had co-occurrence of both conditions. Among those with DT, a higher proportion had thrombocytopenia. Those with seizures had lower blood platelet count and a higher proportion of them had thrombocytopenia. The sensitivity and specificity of thrombocytopenia for the development of DT during the treatment period was 70% and 69%, respectively. The positive predictive value (PPV) was 6% and the negative predictive value (NPV) was 99%. For the development of seizures, the figure for sensitivity was 75% and for specificity 69%. The figures for PPV and NPV were similar as those for the development of DT. CONCLUSIONS: Thrombocytopenia is more frequent in patients who develop severe AWS (DT or seizures). The findings, including the high NPV of thrombocytopenia, must be interpreted with caution due to the small number of patients who developed AWS. Further studies replicating the present finding are therefore needed before the clinical usefulness can be considered. PMID- 19293145 TI - Nicotinic acetylcholine receptor signalling: roles in Alzheimer's disease and amyloid neuroprotection. AB - Alzheimer's disease (AD), the major contributor to dementia in the elderly, involves accumulation in the brain of extracellular plaques containing the beta amyloid protein (Abeta) and intracellular neurofibrillary tangles of hyperphosphorylated tau protein. AD is also characterized by a loss of neurons, particularly those expressing nicotinic acetylcholine receptors (nAChRs), thereby leading to a reduction in nAChR numbers. The Abeta(1-42) protein, which is toxic to neurons, is critical to the onset and progression of AD. The discovery of new drug therapies for AD is likely to be accelerated by an improved understanding of the mechanisms whereby Abeta causes neuronal death. We examine the evidence for a role in Abeta(1-42) toxicity of nAChRs; paradoxically, nAChRs can also protect neurons when activated by nicotinic ligands. Abeta peptides and nicotine differentially activate several intracellular signaling pathways, including the phosphatidylinositol 3-kinase/v-akt murine thymoma viral oncogene homolog pathway, the extracellular signal-regulated kinase/mitogen-activated protein kinase, and JAK-2/STAT-3 pathways. These pathways control cell death or survival and the secretion of Abeta peptides. We propose that understanding the differential activation of these pathways by nicotine and/or Abeta(1-42) may offer the prospect of new routes to therapy for AD. PMID- 19293149 TI - Lysophosphatidic acid 2 receptor-mediated supramolecular complex formation regulates its antiapoptotic effect. AB - The G protein-coupled lysophosphatidic acid 2 (LPA(2)) receptor elicits prosurvival responses to prevent and rescue cells from apoptosis. However, G protein-coupled signals are not sufficient for the full protective effect of LPA(2). LPA(2) differs from other LPA receptor subtypes in the C-terminal tail, where it contains a zinc finger-binding motif for the interactions with LIM domain-containing TRIP6 and proapoptotic Siva-1, and a PDZ-binding motif through which it complexes with the NHERF2 scaffold protein. In this report, we identify a unique CXXC motif of LPA(2) responsible for the binding to TRIP6 and Siva-1, and demonstrate that disruption of these macromolecular complexes or knockdown of TRIP6 or NHERF2 expression attenuates LPA(2)-mediated protection from chemotherapeutic agent-induced apoptosis. In contrast, knockdown of Siva-1 expression enhances this effect. Furthermore, a PDZ-mediated direct interaction between TRIP6 and NHERF2 facilitates their interaction with LPA(2). Together, these results suggest that in addition to G protein-activated signals, the cooperation embedded in the LPA(2)-TRIP6-NHERF2 ternary complex provides a novel ligand-dependent signal amplification mechanism that is required for LPA(2) mediated full activation of antiapoptotic signaling. PMID- 19293150 TI - Overexpression of E-cadherin on melanoma cells inhibits chemokine-promoted invasion involving p190RhoGAP/p120ctn-dependent inactivation of RhoA. AB - Melanoma cells express the chemokine receptor CXCR4 that confers high invasiveness upon binding to its ligand CXCL12. Melanoma cells at initial stages of the disease show reduction or loss of E-cadherin expression, but recovery of its expression is frequently found at advanced phases. We overexpressed E cadherin in the highly invasive BRO lung metastatic cell melanoma cell line to investigate whether it could influence CXCL12-promoted cell invasion. Overexpression of E-cadherin led to defective invasion of melanoma cells across Matrigel and type I collagen in response to CXCL12. A decrease in individual cell migration directionality toward the chemokine and reduced adhesion accounted for the impaired invasion. A p190RhoGAP-dependent inhibition of RhoA activation was responsible for the impairment in chemokine-stimulated E-cadherin melanoma transfectant invasion. Furthermore, we show that p190RhoGAP and p120ctn associated predominantly on the plasma membrane of cells overexpressing E cadherin, and that E-cadherin-bound p120ctn contributed to RhoA inactivation by favoring p190RhoGAP-RhoA association. These results suggest that melanoma cells at advanced stages of the disease could have reduced metastatic potency in response to chemotactic stimuli compared with cells lacking E-cadherin, and the results indicate that p190RhoGAP is a central molecule controlling melanoma cell invasion. PMID- 19293151 TI - Identification of Flightless-I as a substrate of the cytokine-independent survival kinase CISK. AB - Phosphatidylinositol (PI) 3-kinase mediates multiple pathways that regulate many aspects of the cell including metabolism, survival, migration, and proliferation. Both Akt and cytokine-independent survival kinase (CISK)/SGK3 are known AGC family protein kinases that function downstream of PI 3-kinase. Although the Akt signaling pathway has been studied extensively, the specific signaling cascades that are modulated by CISK remain to be elucidated. To understand CISK function, we affinity-purified the CISK protein complex and identified Flightless-I (FLII) as a novel downstream target of CISK. Here we show that FLII is an in vivo substrate of CISK that functions downstream of PI 3-kinase. CISK can associate with FLII and phosphorylate FLII at residues Ser(436) and Thr(818). FLII has been shown to act as a co-activator for nuclear hormone receptors such as estrogen receptor (ER). We demonstrate here that CISK can enhance ER transcription, which is dependent on its kinase activity, and mutation of CISK phosphorylation sites on FLII attenuates its activity as an ER co-activator. Furthermore, FLII knockdown by RNA interference renders 32D cells more sensitive to interleukin-3 withdrawal-induced apoptosis, suggesting that FLII itself is also a survival factor. These findings support the model that CISK phosphorylates FLII and activates nuclear receptor transcription and suggest a new cell survival signaling pathway mediated by PI 3-kinase and CISK. PMID- 19293152 TI - Inhibition of sphingosine kinase by bovine viral diarrhea virus NS3 is crucial for efficient viral replication and cytopathogenesis. AB - Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid implicated in diverse cellular functions including survival, proliferation, tumorigenesis, inflammation, and immunity. Sphingosine kinase (SphK) contributes to these functions by converting sphingosine to S1P. We report here that the nonstructural protein NS3 from bovine viral diarrhea virus (BVDV), a close relative of hepatitis C virus (HCV), binds to and inhibits the catalytic activity of SphK1 independently of its serine protease activity, whereas HCV NS3 does not affect SphK1 activity. Uncleaved NS2-3 from BVDV was also found to interact with and inhibit SphK1. We suspect that inhibition of SphK1 activity by BVDV NS3 and NS2-3 may benefit viral replication, because SphK1 inhibition by small interfering RNA, chemical inhibitor, or overexpression of catalytically inactive SphK1 results in enhanced viral replication, although the mechanisms by which SphK1 inhibition leads to enhanced viral replication remain unknown. A role of SphK1 inhibition in viral cytopathogenesis is also suggested as overexpression of SphK1 significantly attenuates the induction of apoptosis in cells infected with cytopathogenic BVDV. These findings suggest that SphK is targeted by this virus to regulate its catalytic activity. PMID- 19293153 TI - Intermedilysin-receptor interactions during assembly of the pore complex: assembly intermediates increase host cell susceptibility to complement-mediated lysis. AB - Intermedilysin (ILY) is an unusual member of the family of cholesterol-dependent cytolysins because it binds to human CD59 (hCD59) rather than directly to cholesterol-rich membranes. Binding of ILY to hCD59 initiates a series of conformational changes within the toxin that result in the conversion of the soluble monomer into an oligomeric membrane-embedded pore complex. In this study the association of ILY with its membrane receptor has been examined throughout the assembly and formation of the pore complex. Using ILY mutants trapped at various stages of pore assembly, we show ILY remains engaged with hCD59 throughout the assembly of the prepore oligomer, but it disengages from the receptor upon the conversion to the pore complex. We further show that the assembly intermediates increase the sensitivity of the host cell to lysis by its complement membrane attack complex, apparently by blocking the hCD59-binding site for complement proteins C8alpha and C9. PMID- 19293154 TI - Immunoglobulin gene transcripts have distinct VHDJH recombination characteristics in human epithelial cancer cells. AB - It was well accepted that only B-lymphocytes and plasma cells expressed immunoglobulin (Ig) gene. However, our group and others have confirmed that non-B cells, such as epithelial cancer cells, can also express Ig. The aim of this work is to elucidate the role of non-B-cell-derived Ig by investigating the characteristics of the Ig heavy chain (IgH) gene repertoire in epithelial cancer cells. We cloned and sequenced 89 V(H)DJ(H) (V-D-J recombination of the IgH variable region) transcripts by microdissecting cells from eight different types of epithelial cancers and two cancer cell lines (HT-29 and HeLa S3). The cancer derived Ig gene repertoire showed specific restricted patterns of V(H)DJ(H) recombination with seven sets of predominant V(H)DJ(H) sequences. Surprisingly, within a set of V(H)DJ(H) recombination, the variable (V) sequences derived from different cancer types had not only identical heavy chain variable (VH), diversity (D), and joining (JH) segments usage, but also identical junctions and mutation targets in the V(H) region. The V(Hgamma)DJ(Hgamma) (but not V(Hmicro)DJ(Hmicro)) in the cancer-derived sequences had a high mutation rate; however, it was shown that the mechanism of hypermutation was different from antigen selection in B-cell-derived V(Hgamma)DJ(Hgamma)sequences. In contrast to V(Hmicro)DJ(Hmicro), the V(Hgamma)DJ(Hgamma) sequences did not appear to originate from classical class switching. These results suggest that cancer derived Ig genes have a distinct repertoire that may have implications for their role in carcinogenesis. PMID- 19293155 TI - Oxidizable residues mediating protein stability and cytoprotective interaction of DJ-1 with apoptosis signal-regulating kinase 1. AB - Parkinson disease (PD)-associated genomic deletions and the destabilizing L166P point mutation lead to loss of the cytoprotective DJ-1 protein. The effects of other PD-associated point mutations are less clear. Here we demonstrate that the M26I mutation reduces DJ-1 expression, particularly in a null background (knockout mouse embryonic fibroblasts). Thus, homozygous M26I mutation causes loss of DJ-1 protein. To determine the cellular consequences, we measured suppression of apoptosis signal-regulating kinase 1 (ASK1) and cytotoxicity for [M26I]DJ-1, and systematically all other DJ-1 methionine and cysteine mutants. C106A mutation of the central redox site specifically abolished binding to ASK1 and the cytoprotective activity of DJ-1. DJ-1 was apparently recruited into the ASK1 signalosome via Cys-106-linked mixed disulfides. The designed higher order oxidation mimicking [C106DD]DJ-1 non-covalently bound to ASK1 even in the absence of hydrogen peroxide and conferred partial cytoprotection. Interestingly, mutations of peripheral redox sites (C46A and C53A) and M26I also led to constitutive ASK1 binding. Cytoprotective [wt]DJ-1 bound to the ASK1 N terminus (which is known to bind another negative regulator, thioredoxin 1), whereas [M26I]DJ-1 bound to aberrant C-terminal site(s). Consequently, the peripheral cysteine mutants retained cytoprotective activity, whereas the PD-associated mutant [M26I]DJ-1 failed to suppress ASK1 activity and nuclear export of the death domain-associated protein Daxx and did not promote cytoprotection. Thus, cytoprotective binding of DJ-1 to ASK1 depends on the central redox-sensitive Cys 106 and may be modulated by peripheral cysteine residues. We suggest that impairments in oxidative conformation changes of DJ-1 might contribute to PD neurodegeneration. PMID- 19293156 TI - Determination of in vivo dissociation constant, KD, of Cdc42-effector complexes in live mammalian cells using single wavelength fluorescence cross-correlation spectroscopy. AB - The RhoGTPase Cdc42 coordinates cell morphogenesis, cell cycle, and cell polarity decisions downstream of membrane-bound receptors through distinct effector pathways. Cdc42-effector protein interactions represent important elements of cell signaling pathways that regulate cell biology in systems as diverse as yeast and humans. To derive mechanistic insights into cell signaling pathways, it is vital that we generate quantitative data from in vivo systems. We need to be able to measure parameters such as protein concentrations, rates of diffusion, and dissociation constants (K(D)) of protein-protein interactions in vivo. Here we show how single wavelength fluorescence cross-correlation spectroscopy in combination with Forster resonance energy transfer analysis can be used to determine K(D) of Cdc42-effector interactions in live mammalian cells. Constructs encoding green fluorescent protein or monomeric red fluorescent protein fusion proteins of Cdc42, an effector domain (CRIB), and two effectors, neural Wiskott Aldrich syndrome protein (N-WASP) and insulin receptor substrate protein (IRSp53), were expressed as pairs in Chinese hamster ovary cells, and concentrations of free protein as well as complexed protein were determined. The measured K(D) for Cdc42V12-N-WASP, Cdc42V12-CRIB, and Cdc42V12-IRSp53 was 27, 250, and 391 nm, respectively. The determination of K(D) for Cdc42-effector interactions opens the way to describe cell signaling pathways quantitatively in vivo in mammalian cells. PMID- 19293157 TI - The membrane-binding motif of the chloroplast signal recognition particle receptor (cpFtsY) regulates GTPase activity. AB - The chloroplast signal recognition particle (cpSRP) and its receptor (cpFtsY) function in thylakoid biogenesis to target integral membrane proteins to thylakoids. Unlike cytosolic SRP receptors in eukaryotes, cpFtsY partitions between thylakoid membranes and the soluble stroma. Based on sequence alignments, a membrane-binding motif identified in Escherichia coli FtsY appears to be conserved in cpFtsY, yet whether the proposed motif is responsible for the membrane-binding function of cpFtsY has yet to be shown experimentally. Our studies show that a small N-terminal region in cpFtsY stabilizes a membrane interaction critical to cpFtsY function in cpSRP-dependent protein targeting. This membrane-binding motif is both necessary and sufficient to direct cpFtsY and fused passenger proteins to thylakoids. Our results demonstrate that the cpFtsY membrane-binding motif may be functionally replaced by the corresponding region from E. coli, confirming that the membrane-binding motif is conserved among organellar and prokaryotic homologs. Furthermore, the capacity of cpFtsY for lipid binding correlates with liposome-induced GTP hydrolysis stimulation. Mutations that debilitate the membrane-binding motif in cpFtsY result in higher rates of GTP hydrolysis, suggesting that negative regulation is provided by the intact membrane-binding region in the absence of a bilayer. Furthermore, NMR and CD structural studies of the N-terminal region and the analogous region in the E. coli SRP receptor revealed a conformational change in secondary structure that takes place upon lipid binding. These studies suggest that the cpFtsY membrane binding motif plays a critical role in the intramolecular communication that regulates cpSRP receptor functions at the membrane. PMID- 19293158 TI - Molecular basis for Bcl-2 homology 3 domain recognition in the Bcl-2 protein family: identification of conserved hot spot interactions. AB - The proteins of the Bcl-2 family are important regulators of apoptosis, or programmed cell death. These proteins regulate this fundamental biological process via the formation of heterodimers involving both pro- and anti-apoptotic family members. Disruption of the balance between anti- and pro-apoptotic Bcl-2 proteins is the cause of numerous pathologies. Bcl-xl, an anti-apoptotic protein of this family, is known to form heterodimers with multiple pro-apoptotic proteins, such as Bad, Bim, Bak, and Bid. To elucidate the molecular basis of this recognition process, we used molecular dynamics simulations coupled with the Molecular Mechanics/Poisson-Boltzmann Surface Area approach to identify the amino acids that make significant energetic contributions to the binding free energy of four complexes formed between Bcl-xl and pro-apoptotic Bcl-2 homology 3 peptides. A fifth protein-peptide complex composed of another anti-apoptotic protein, Bcl w, in complex with the peptide from Bim was also studied. The results identified amino acids of both the anti-apoptotic proteins as well as the Bcl-2 homology 3 (BH3) domains of the pro-apoptotic proteins that make strong, recurrent interactions in the protein complexes. The calculations show that the two anti apoptotic proteins, Bcl-xl and Bcl-w, share a similar recognition mechanism. Our results provide insight into the molecular basis for the promiscuous nature of this molecular recognition process by members of the Bcl-2 protein family. These amino acids could be targeted in the design of new mimetics that serve as scaffolds for new antitumoral molecules. PMID- 19293160 TI - Patient-reported outcomes improve with etanercept plus methotrexate in active early rheumatoid arthritis and the improvement is strongly associated with remission: the COMET trial. AB - OBJECTIVES: To compare the effects of etanercept (ETN) 50 mg once weekly plus methotrexate (MTX) versus MTX alone on patient-reported outcomes (PROs) and the relationship between remission and PRO improvement. METHODS: In this double blind, randomised clinical trial (COMET), PROs included: the Health Assessment Questionnaire (HAQ), EuroQoL health status, fatigue and pain visual analogue scales, Hospital Anxiety and Depression Scale, and Medical Outcomes Short-Form 36. Mean changes from baseline were analysed by analysis of covariance using the last observation carried forward method. Results from week 52 are presented. RESULTS: Most PROs demonstrated significantly greater improvements with ETN+MTX than MTX alone, including physical functioning, pain, fatigue and overall health status. A significantly greater improvement in HAQ score was observed in the ETN+MTX than the MTX group (-1.02 vs -0.72; p<0.001) and a greater proportion reached the minimal clinically important difference of 0.22 (88% vs 78%; p<0.006). The relationship between PRO score and clinical status indicated that improvement was greatest among patients achieving remission. CONCLUSIONS: Early treatment with ETN+MTX leads to significantly greater improvements in multiple dimensions of PROs than MTX alone. The close relationship between disease activity and PRO improvement suggests that early treatment, with remission as a goal, should maximise the chance of restoring normal functioning and HRQoL. PMID- 19293161 TI - Anti-inflammatory and cartilage-protecting effects of an intra-articularly injected anti-TNF{alpha} single-chain Fv antibody (ESBA105) designed for local therapeutic use. AB - OBJECTIVES: (1) To show that a single-chain Fv antibody (scFv) against tumour necrosis factor alpha (TNFalpha) (ESBA105) has efficacy comparable to a full length anti-TNFalpha IgG (infliximab); (2) to evaluate whether ESBA105 has all the properties required for the local treatment of arthritis; and (3) to investigate its discriminative tissue penetration properties. METHODS: In vivo efficacy was measured in arthritis of the knee joint induced by the intra articular injection of recombinant human TNFalpha (rhTNFalpha) in Lewis rats. Cartilage penetration of scFv (ESBA105) and full length IgG (infliximab) were studied in bovine cartilage specimens ex vivo. Tissue penetration, biodistribution and pharmacokinetics of ESBA105 were followed and compared after intra-articular and intravenous administration. RESULTS: In cell culture, ESBA105 showed similar TNFalpha inhibitory potency to infliximab. In vivo, ESBA105 inhibited rhTNFalpha-induced synovial inflammation in rats with efficacy again comparable to infliximab. An 11-fold molar excess of ESBA105 over rhTNFalpha resulted in 90% inhibition of knee joint swelling, inflammatory infiltrates and proteoglycan loss from cartilage. In ex vivo studies of bovine cartilage, ESBA105 penetrated well into the cartilage whereas infliximab remained on the surface. In vivo, rapid penetration into the synovial tissue, cartilage and surrounding tissues was observed following intra-articular injection of [(125)I]-ESBA105 into the knee joint of rabbits. CONCLUSIONS: ESBA105 potently inhibits inflammation and prevents cartilage damage triggered by TNFalpha. In contrast to a full length IgG, ESBA105 also penetrates into cartilage and can be expected to reverse the TNFalpha-induced catabolic state of articular cartilage in arthritides. PMID- 19293162 TI - Antifibroblast antibodies from systemic sclerosis patients bind to {alpha} enolase and are associated with interstitial lung disease. AB - OBJECTIVE: To identify target antigens of antifibroblast antibodies (AFA) in systemic sclerosis (SSc) patients. PATIENTS AND METHODS: In the first part, sera from 24 SSc patients (12 with pulmonary arterial hypertension (PAH) and 12 without) and 36 idiopathic PAH patients, tested in pooled sera for groups of three, were compared with a sera pool from 14 healthy controls (HC). Serum IgG reactivity was analysed by the use of a two-dimensional electrophoresis and immunoblotting technique with normal human fibroblasts antigens. In the second part, serum IgG reactivity for two groups: 158 SSc, 67 idiopathic PAH and 100 HC; and 35 SSc and 50 HC was tested against alpha-enolase from Saccharomyces cerevisiae and recombinant human (rHu) alpha-enolase, respectively, on ELISA. RESULTS: In the first part, alpha-enolase was identified as a main target antigen of AFA from SSc patients. In the second part, 37/158 (23%) SSc patients, 6/67 (9%) idiopathic PAH patients and 4/100 (4%) HC (p<0.001) had anti-S cerevisiae alpha-enolase antibodies; 12/35 (34%) SSc patients and 3/50 (6%) HC had anti-rHu alpha-enolase antibodies (p = 0.001). In SSc, the presence of anti-S cerevisiae alpha-enolase antibodies was associated with interstitial lung disease (ILD), decreased total lung capacity (73.2% vs 89.7%; p<0.001) and diffusion capacity for carbon monoxide (47.4% vs 62.3%; p<0.001), and antitopoisomerase 1 antibodies (46% vs 21%; p = 0.005) but not anticentromere antibodies (11% vs 34%; p = 0.006). Results were similar with rHu alpha-enolase testing. CONCLUSION: In SSc, AFA recognise alpha-enolase and are associated with ILD and antitopoisomerase antibodies. PMID- 19293163 TI - Penetration of topical and subconjunctival corticosteroids into human aqueous humour and its therapeutic significance. AB - Topical and subconjunctival corticosteroids are some of the most effective and compelling treatment options in ocular inflammatory diseases. A systematic review of literature indexed by Ovid MEDLINE & EMBASE was performed up to December 2008. There are few studies on their aqueous penetration in human subjects. This review article discusses the penetration of different ocular corticosteroids into human aqueous humour along with the therapeutic implications on management of ocular surface diseases, immune-related corneal diseases, anterior uveitis and postoperative anti-inflammatory use. In the context of the paucity of well constructed, prospective clinical trials comparing the efficacy of different corticosteroids, it provides guiding principles for the use of topical corticosteroids. Dexamethasone alcohol 0.1% and prednisolone acetate 1% are potent corticosteroids, but the latter achieves the highest aqueous concentration within 2 h and maintains higher levels for 24 h. Subconjunctival corticosteroids provide very high concentrations in the aqueous which maintain higher concentrations for longer periods. PMID- 19293164 TI - Exercise prescription for the overweight and the obese: how to quantify and yet keep it simple. AB - An activity point system (APS) is presented that assists healthcare providers and health professionals with a way to prescribe exercise in a semiquantitative manner and to motivate their patients to begin an exercise programme. Activity points are specific to one's body weight, body mass index and activity intensity for 10-minute bouts of activity performed. With a goal of accumulating 100-300 activity points per week, the APS provides a simple yet quantitatively accurate way to prescribe exercise for overweight and obese adults. PMID- 19293165 TI - Vascularity and tendon pathology in the rotator cuff: a review of literature and implications for rehabilitation and surgery. AB - OBJECTIVE: To compile histological and imaging research detailing the microvascularity of the rotator cuff and determine the clinical application of these findings for clinicians. METHODS: A computer-assisted literature search of MEDLINE (1966 to September 2008) using keywords related to blood flow to the shoulder and limited to humans and English language. A hand search was also performed by three of the authors. RESULTS: Nineteen studies met inclusion and exclusion criteria. CONCLUSIONS: The relationship between the variables of vascularity, age and degeneration remains unclear. Recent studies with stronger design and better technology support the fact that increased vascularity is a normal response to smaller tears, but that as tear size increases the healing response fails and decreased vascularity is observed. Also, impingement may cause hypovascularity. These studies support the possibility that people without symptoms may have normal blood flow even with ageing. Finally, exercise may increase blood flow to the rotator cuff. These findings have both surgical and rehabilitation implications. PMID- 19293166 TI - Traffic exposure and subclinical cardiovascular disease: is the association modified by socioeconomic characteristics of individuals and neighbourhoods? Results from a multilevel study in an urban region. AB - OBJECTIVES: Traffic-related pollution is associated with cardiovascular disease in general, but previous studies suggested that low socioeconomic status (SES) groups might be more susceptible towards a negative impact. We examined whether the association between long-term exposure to high traffic and early signs of coronary artery disease is modified by SES. METHODS: Individual-level medical and social data from a population-based study were linked with census information on neighbourhood socioeconomic characteristics. Residential exposure to traffic was defined as proximity to major roads using a geographical information system. We studied associations between high traffic and coronary artery calcification (CAC) within strata of SES to examine effect modification. Data stem from an epidemiological study in Germany including 2264 women and 2037 men (45-75 years). RESULTS: High traffic and low SES were both associated with higher amounts of calcification (>or=75th age-specific percentile). More participants with low SES lived close to major roads while stratified analyses did not indicate higher susceptibility in low SES groups. Participants with low SES and simultaneous exposure to high traffic had highest levels of CAC. For example, the prevalence of high calcification was 23.9% in better-educated men with low traffic exposure but 37.7% in lower-educated men with high traffic exposure (women: 22.0% vs 28.1%). CONCLUSIONS: High traffic exposure was associated with coronary calcification in all social groups, but as low SES individuals had higher calcification in general and were also more often exposed to traffic, existing inequalities could be further shaped by traffic exposure. PMID- 19293167 TI - Social inequality in the prevalence of depressive disorders. AB - BACKGROUND: Uncertainties exist about the strength of the relation between socioeconomic position and depressive disorders. The aim of this study was to investigate the association between education, occupation, employment and income and depressive disorders measured as minor and major depression, as well as antidepressant prescriptions. METHODS: Data were collected from a Danish cross sectional study collected year 2000, comprising 9254 subjects, 55% women, and aged 36-56 years. Register-based information on education, income and prescription were used. RESULTS: The prevalence of major depression DSM-IV algorithm was 3.3% among men and women, whereas minor depression and prescriptions revealed statistically significant higher prevalence among females. A social gradient was found for all depressive end-points with the strongest estimates related to major depressive disorder (MDD). The associations were as follows: MDD and low education odds ratio (OR) 2.38 (CI 95% 1.68 to 3.37), MDD and non-employment OR 11.67 (CI 95% 8.06 to 16.89), MDD and low income OR 9.78 (CI 95% 6.49 to 14.74). Education only explained a minor part of the association between non-employment and depressive disorders and no associations were found between education and prescription. This indicates a strong two-way association between depression and non-employment, low-income respectively. CONCLUSION: A social gradient in depressive disorders was found regardless of socioeconomic position being measured by education, occupation, employment or income. Severe socioeconomic consequences of depression are indicated by the fact that the associations with non-employment and low income were much stronger than the association with low education. PMID- 19293168 TI - Genetics of microtia and associated syndromes. AB - Microtia is a congenital anomaly, characterised by a small, abnormally shaped auricle (pinna). It is usually accompanied by a narrow, blocked or absent ear canal. Microtia can occur as the only clinical abnormality or as part of a syndrome. The estimated prevalence of microtia is 0.8-4.2 per 10 000 births, and it is more common in men. Microtia can have a genetic or environmental predisposition. Mendelian hereditary forms of microtia with an autosomal dominant or recessive mode of inheritance, and some forms due to chromosomal aberrations have been reported. Several responsible genes have been identified, most of them being homeobox genes. Mouse models have been very useful to study these genes, providing valuable information on the development of the auditory system. In this article, we review the epidemiological characteristics of microtia and the environmental causes involved. In addition, we discuss the development of the auditory system, specifically the relevant aspects of external and middle ear development. The focus of this review is to discuss the genetic aspects of microtia and associated syndromes. The clinical aspects of various disorders involving microtia are also discussed in relation to the genes that are causing them. PMID- 19293169 TI - U-type exchange is the most frequent mechanism for inverted duplication with terminal deletion rearrangements. AB - BACKGROUND: Chromosomal rearrangements resulting in an interstitial inverted duplication with concomitant terminal deletion were first described for the short arm of chromosome 8 in 1976. Since then, this type of alteration has been identified and characterised for most chromosome arms. Three mechanisms are commonly proposed to explain the origin of this type of rearrangement. All three mechanisms involve formation of a dicentric chromosome that then breaks in a subsequent meiotic division to produce a monocentric duplicated and deleted chromosome. However, the events leading to the formation of the dicentric chromosome differ between the mechanisms. In one mechanism, either parent carries a paracentric inversion. This results in formation of a loop during meiotic pairing with a recombination event occurring in the loop. In the second mechanism, inverted low copy repeats in the same chromosome arm allow partial folding of one homologue onto itself with a recombination event between the inverted repeats. The third mechanism involves a pre-meiotic double-strand break with subsequent fusion, or U-type exchange, between the sister chromatids. The first two mechanisms require a single copy region to exist between the duplicated and deleted regions on the derivative chromosome, and therefore high resolution analysis of the rearrangement can be used to distinguish between these mechanisms. METHODS AND RESULTS: Using G-banded chromosome analysis, fluorescence in situ hybridisation (FISH) and array comparative genomic hybridisation (CGH), we describe 17 new cases of inverted duplication with terminal deletion of 2q, 4p, 5p, 6q, 8p, 9p, 10q, 13q, 15q, 18p, 18q, and 22q. CONCLUSIONS: These new cases, combined with previously described cases, demonstrate that U-type exchange is the most frequent mechanism for this rearrangement and can be observed on most, or perhaps all, chromosome arms. PMID- 19293170 TI - Rhabdomyosarcoma in patients with constitutional mismatch-repair-deficiency syndrome. AB - BACKGROUND: Biallelic germline mutations in the mismatch repair genes MLH1, MSH2, MSH6 or PMS2 cause a recessive childhood cancer syndrome characterised by early onset malignancies and signs reminiscent of neurofibromatosis type 1 (NF1). Alluding to the underlying genetic defect, we refer to this syndrome as constitutional mismatch repair-deficiency (CMMR-D) syndrome. The tumour spectrum of CMMR-D syndrome includes haematological neoplasias, brain tumours and Lynch syndrome-associated tumours. Other tumours, such as neuroblastoma, Wilm tumour, ovarian neuroectodermal tumour or infantile myofibromatosis, have so far been found only in individual cases. RESULTS: We analysed two consanguineous families that had members with suspected CMMR-D syndrome who developed rhabdomyosarcoma among other neoplasias. In the first family, we identified a pathogenic PMS2 mutation for which the affected patient was homozygous. In family 2, immunohistochemistry analysis showed isolated loss of PMS2 expression in all tumours in the affected patients, including rhabdomyosarcoma itself and the surrounding normal tissue. Together with the family history and microsatellite instability observed in one tumour this strongly suggests an underlying PMS2 alteration in family 2 also. CONCLUSION: Together, these two new cases show that rhabdomyosarcoma and possibly other embryonic tumours, such as neuroblastoma and Wilm tumour, belong to the tumour spectrum of CMMR-D syndrome. Given the clinical overlap of CMMR-D syndrome with NF1, we suggest careful examination of the family history in patients with embryonic tumours and signs of NF1 as well as analysis of the tumours for loss of one of the mismatch repair genes and microsatellite instability. Subsequent mutation analysis will lead to a definitive diagnosis of the underlying disorder. PMID- 19293171 TI - Effect of mannitol and hypertonic saline on cerebral oxygenation in patients with severe traumatic brain injury and refractory intracranial hypertension. AB - BACKGROUND: The impact of osmotic therapies on brain oxygen has not been extensively studied in humans. We examined the effects on brain tissue oxygen tension (PbtO(2)) of mannitol and hypertonic saline (HTS) in patients with severe traumatic brain injury (TBI) and refractory intracranial hypertension. METHODS: 12 consecutive patients with severe TBI who underwent intracranial pressure (ICP) and PbtO(2) monitoring were studied. Patients were treated with mannitol (25%, 0.75 g/kg) for episodes of elevated ICP (>20 mm Hg) or HTS (7.5%, 250 ml) if ICP was not controlled with mannitol. PbtO(2), ICP, mean arterial pressure, cerebral perfusion pressure (CPP), central venous pressure and cardiac output were monitored continuously. RESULTS: 42 episodes of intracranial hypertension, treated with mannitol (n = 28 boluses) or HTS (n = 14 boluses), were analysed. HTS treatment was associated with an increase in PbtO(2) (from baseline 28.3 (13.8) mm Hg to 34.9 (18.2) mm Hg at 30 min, 37.0 (17.6) mm Hg at 60 min and 41.4 (17.7) mm Hg at 120 min; all p<0.01) while mannitol did not affect PbtO(2) (baseline 30.4 (11.4) vs 28.7 (13.5) vs 28.4 (10.6) vs 27.5 (9.9) mm Hg; all p>0.1). Compared with mannitol, HTS was associated with lower ICP and higher CPP and cardiac output. CONCLUSIONS: In patients with severe TBI and elevated ICP refractory to previous mannitol treatment, 7.5% hypertonic saline administered as second tier therapy is associated with a significant increase in brain oxygenation, and improved cerebral and systemic haemodynamics. PMID- 19293172 TI - Misplacing objects in mild to moderate Alzheimer's disease: a descriptive analysis from the VISTA clinical trial. AB - BACKGROUND: Misplacing objects is a commonly reported symptom of Alzheimer's disease (AD) but it is little described systematically and conflicting characterisations (losing/forgetting the location of objects versus inappropriate placement) of this "warning sign" of dementia currently exist. Patient and carer descriptions of misplacing in clinical interviews are reported here. METHODS: This was a secondary qualitative analysis of video recorded, open ended and semistructured clinical interviews with 130 community dwelling patients with mild to moderate AD (mean age 77 (7.7) years; 63% women; 67% mild AD) and their carers who participated in the Video Imaging Synthesis of Treating Alzheimer's Disease (VISTA) study, a 4 month, randomised, placebo controlled trial of galantamine. Employing a framework analysis approach, we summarised descriptive accounts of misplacing with relevant proportions. RESULTS: Recurrent incidents of misplacing were described for 96/130 (74%) study patients, 45 of whom established treatment goals to track this problem. For most (78/96, 81%), misplacing was the inability to recall where an item had been set down or put away. Fewer patients (25/96, including 18 with recall misplacing) put objects in unusual or incorrect places. Patients were commonly aware of their misplacing (56/96, 58%) and were distressed by it (31/56). Patients who misplaced also displayed tendencies towards delusions/hallucinations (51/96, but only directly related to misplacing in 17 cases) and hiding items (15%) CONCLUSION: Misplacing is a common phenomenon in mild to moderate AD. Here, misplacing was usually described as an inability to recall where an item was set down, more so than the inappropriate placement of items. PMID- 19293173 TI - Prospective surveillance of drain associated meningitis/ventriculitis in a neurosurgery and neurological intensive care unit. AB - OBJECTIVES: There are currently no data available on drain associated infection occurrence related to the number of drainage days (DD), and thus drain associated infection rates. Therefore, a prospective surveillance study was conducted to determine drain associated infection rates and DD of hospital acquired external ventricular drain (EVD) and lumbar drain (LD) associated meningitis/ventriculitis in a neurosurgery and a neurological intensive care unit. METHODS: All patients admitted in 2005 and 2006 were documented. Data on age, admitting diagnosis, type and duration of drain, duration of hospital stay and occurrence of meningitis were recorded and analysed statistically. RESULTS: A total of 1333 patients were included, amounting to 3023 DD. After exclusion of 15 contaminations, a total of 26 cases of meningitis were reported accounting for an overall device associated meningitis rate of 8.6 infections/1000 DD. Infections associated with LD seemed to occur more frequently (19.9/1000 DD) compared with EVD (6.3/1000 DD). The presence of intraventricular blood and previous trauma were significant risk factors for infection (p = 0.003; p = 0.04). Finally, length of stay was significantly longer in meningitis patients (p = 0.0003). Coagulase negative staphylococci were the main pathogen (56%) causing meningitis, followed by Staphylococcus aureus (25%). CONCLUSIONS: To the best of the authors' knowledge, this study represents the first to provide data on EVD as well as LD associated meningitis rates calculated per 1000 DD; a parameter that is well established for other invasive devices such as central venous and urinary tract catheters. However, further prospective studies are needed to investigate the possible risk factors for meningitis. PMID- 19293174 TI - Perioperative risk factors for short term shunt revisions in adult hydrocephalus patients. AB - OBJECTIVES: The aim of this study was to prospectively study perioperative variables associated with revision after shunt surgery for adult hydrocephalus. METHODS: Two protocols were designed to prospectively study perioperative risk factors during shunt insertion. Over 10 years (1995-2004), 450 adult (age >16 years) patients with first time shunt implantations were studied. Patients who had been treated with endoscopic third ventriculostomy were excluded from the study. All shunts were designated as meeting one of two end points: (1) shunt failure requiring revision within 6 months or (2) no shunt failure within 6 months. Shunt revision within 6 months postoperatively was considered to be related to the shunting procedure. RESULTS: 85 shunt revisions were performed within 6 months after insertion. During the study period the revision rate declined from 21.1% to 9.1%. Revision rates were the same for ventriculoperitoneal (n = 411) and ventriculoatrial (n = 39) shunts. The predictive values of variables related to the patient, operating room, surgical technique and shunt system were analysed to determine shunt outcome. CONCLUSIONS: Right frontal placement of the ventricular catheter was associated with the lowest rate of revisions. Adjustable valves were associated with a lower risk for shunt revision. Shunt revision rates did not differ between ventriculoperitoneal and ventriculoatrial shunts. PMID- 19293175 TI - Clifton E. Barry, III: TB's strategic opponent. Interviewed by Amy Maxmen. PMID- 19293176 TI - Formation of vitamin A lipid droplets in pancreatic stellate cells requires albumin. AB - OBJECTIVE: Quiescent pancreatic stellate cells (PSCs) store vitamin A as cytoplasmic lipid droplets, and, when activated by profibrogenic stimuli, they transform into myofibroblast-like cells characterised by the loss of vitamin A droplets. Activation of stellate cells is central to fibrogenesis, but the mechanism for the formation of vitamin A droplets and its relationship to stellate cell activation remain unclear. METHODS: With use of cultured PSCs, an attempt was made to characterise the function of albumin endogenously expressed in stellate cells. RESULTS: Albumin is endogenously expressed in quiescent PSCs, localised in cytoplasmic lipid droplets, and its levels are markedly reduced after stellate cell activation. Continuous albumin expression in stellate cells is sufficient to maintain their fat-storing phenotype even after cell passages and renders cells resistant to the activating effects of transforming growth factor beta (TGFbeta). Forced expression of albumin in PSCs after passage 2 (activated PSCs) induced the re-appearance of lipid droplets and phenotypic changes, which were previously reported with retinol treatment. Retinol increases albumin synthesis in activated PSCs and the suppression of albumin expression using small interfering RNA (siRNA) abolishes retinol-induced effects. CONCLUSIONS: The data demonstrate a novel role for albumin in the formation of cytoplasmic vitamin A lipid droplets in stellate cells, and suggest that albumin may have a direct influence on stellate cell activation. PMID- 19293177 TI - Screening colonoscopy for the detection of neoplastic lesions in asymptomatic HIV infected subjects. AB - BACKGROUND: Although non-AIDS defining malignancies are rapidly increasing as HIV infected subjects live longer, little is know about the results of screening for colonic neoplasms (adenomatous polyps and adenocarcinomas) in this population. METHODS: We conducted a screening colonoscopy study to determine the prevalence of colonic neoplasms in 136 asymptomatic HIV-infected subjects >or=50 years of age and 272 asymptomatic uninfected control subjects matched for age, sex, and family history of colorectal cancer. Advanced neoplasms were defined as adenomas >or=10 mm or any adenoma, regardless of size, with villous histology, high-grade dysplasia, or adenocarcinoma. RESULTS: The prevalence of neoplastic lesions was significantly higher in HIV-infected subjects than in control subjects (62.5% vs 41.2%, p<0.001), and remained highly significant after adjustment for potential confounding variables (odds ratio = 3.00; 95% confidence interval, 1.83 to 4.93). Among patients with colorectal adenocarcinoma, HIV-infected subjects were significantly younger (52.4 (SD 1.3) vs 60.3 (SD 4.0) years, p = 0.002) and were more likely to have advanced cancers (stage III or IV) than control subjects (60.0% vs 16.7%, p = 0.24). Of HIV-infected subjects with advanced neoplasms proximal to the splenic flexure, distal neoplastic lesions were absent in 88.9% of individuals and these would have been missed by flexible sigmoidoscopy. CONCLUSIONS: HIV-infected subjects have a higher prevalence of colonic neoplasms, and adenocarcinomas develop at a younger age and are more advanced than in uninfected subjects. Our findings suggest that screening colonoscopy should be offered to HIV-infected subjects, but the age of initiation and the optimal frequency of screening require further study. PMID- 19293178 TI - Beyond Incrementalism? SCHIP and the politics of health reform. AB - When Congress enacted the State Children's Health Insurance Program (SCHIP) in 1997, it was heralded as a model of bipartisan, incremental health policy. However, despite the program's achievements in the ensuing decade, SCHIP's reauthorization triggered political conflict, and efforts to expand the program stalemated in 2007. The 2008 elections broke that stalemate, and in 2009 the new Congress passed, and President Barack Obama signed, legislation reauthorizing SCHIP. Now that attention is turning to comprehensive health reform, what lessons can reformers learn from SCHIP's political adventures? PMID- 19293179 TI - SLC45A3-ELK4 is a novel and frequent erythroblast transformation-specific fusion transcript in prostate cancer. AB - Chromosomal rearrangements account for all erythroblast transformation-specific (ETS) family member gene fusions that have been reported in prostate cancer and have clinical, diagnostic, and prognostic implications. Androgen-regulated genes account for the majority of the 5' genomic regulatory promoter elements fused with ETS genes. TMPRSS2-ERG, TMPRSS2-ETV1, and SLC45A3-ERG rearrangements account for roughly 90% of ETS fusion prostate cancer. ELK4, another ETS family member, is androgen regulated, involved in promoting cell growth, and highly expressed in a subset of prostate cancer, yet the mechanism of ELK4 overexpression is unknown. In this study, we identified a novel ETS family fusion transcript, SLC45A3-ELK4, and found it to be expressed in both benign prostate tissue and prostate cancer. We found high levels of SLC45A3-ELK4 mRNA restricted to a subset of prostate cancer samples. SLC45A3-ELK4 transcript can be detected at high levels in urine samples from men at risk for prostate cancer. Characterization of the fusion mRNA revealed a major variant in which SLC45A3 exon 1 is fused to ELK4 exon 2. Based on quantitative PCR analyses of DNA, unlike other ETS fusions described in prostate cancer, the expression of SLC45A3-ELK4 mRNA is not exclusive to cases harboring a chromosomal rearrangement. Treatment of LNCaP cancer cells with a synthetic androgen (R1881) revealed that SLC45A3-ELK4, and not endogenous ELK4, mRNA expression is androgen regulated. Altogether, our findings show that SLC45A3 ELK4 mRNA expression is heterogeneous, highly induced in a subset of prostate cancers, androgen regulated, and most commonly occurs through a mechanism other than chromosomal rearrangement (e.g., trans-splicing). PMID- 19293180 TI - Heterozygosity for hypoxia inducible factor 1alpha decreases the incidence of thymic lymphomas in a p53 mutant mouse model. AB - Hypoxia inducible factors (HIF) are critical mediators of the cellular response to decreased oxygen tension and are overexpressed in a number of tumors. Although HIF1alpha and HIF2alpha share a high degree of sequence homology, recent work has shown that the two alpha subunits can have contrasting and tissue-specific effects on tumor growth. To directly compare the role of each HIFalpha subunit in spontaneous tumorigenesis, we bred a mouse model of expanded HIF2alpha expression and Hif1alpha(+/-) mice to homozygotes for the R270H mutation in p53. Here, we report that p53(R270H/R270H) mice, which have not been previously described, develop a unique tumor spectrum relative to p53(R270H/-) mice, including a high incidence of thymic lymphomas. Heterozygosity for Hif1alpha significantly reduced the incidence of thymic lymphomas observed in this model. Moreover, reduced Hif1alpha levels correlated with decreased stabilization of activated Notch1 and expression of the Notch target genes, Dtx1 and Nrarp. These observations uncover a novel role for HIF1alpha in Notch pathway activation during T-cell lymphomagenesis. PMID- 19293181 TI - Circulating B-cell chronic lymphocytic leukemia cells display impaired migration to lymph nodes and bone marrow. AB - Homing to secondary lymphoid organs and bone marrow (BM) is a central aspect of leukemic pathophysiology. We investigated the roles of the two major lymphocyte integrins LFA-1 and VLA-4 on B-cell chronic lymphocytic leukemia (CLL) cells in these processes. We found that the majority of CLL cells expressed significantly reduced LFA-1 due to low beta2 integrin transcripts. VLA-4 expression was heterogeneous but underwent rapid activation by the BM chemokine CXCL12. CLL cells failed to transmigrate across VCAM-1-expressing, ICAM-1-expressing, and CXCL12-expressing endothelium, whereas when LFA-1 expression was regained in subsets of CLL cells, these lymphocytes rapidly transmigrated the endothelium. Furthermore, when injected into tail veins of immunodeficient mice, normal B cells rapidly homed to lymph nodes (LN) in a LFA-1-dependent manner, whereas CLL cells did not. Nevertheless, only residual CLL subsets could reenter BM, whereas both normal and CLL cells homed to the mice spleen in an LFA-1-independent and VLA-4-independent manner. Our results suggest that CLL cells have a reduced capacity to adhere and transmigrate through multiple vascular endothelial beds and poorly home to lymphoid organs other than spleen. Integrin blocking could thus be an efficient strategy to prevent circulating CLL cells from reaching prosurvival niches in LNs and BM but not in spleen. PMID- 19293182 TI - A novel human AlkB homologue, ALKBH8, contributes to human bladder cancer progression. AB - We recently identified a novel human AlkB homologue, ALKBH8, which is expressed in various types of human cancers including human urothelial carcinomas. In examining the role and function of ALKBH8 in human bladder cancer development in vitro, we found that silencing of ALKBH8 through small interfering RNA transfection reduced reactive oxygen species (ROS) production via down-regulation of NAD(P)H oxidase-1 (NOX-1) and induced apoptosis through subsequent activation of c-jun NH(2)-terminal kinase (JNK) and p38. However, we also found that JNK and p38 activation resulted in phosphorylation of H2AX (gammaH2AX), a variant of mammalian histone H2A, which contributes to the apoptosis induced by silencing ALKBH8 and NOX-1. Silencing of ALKBH8 significantly suppressed invasion, angiogenesis, and growth of bladder cancers in vivo as assessed both in the chorioallantoic membrane assay and in an orthotopic mouse model using green fluorescent protein-labeled KU7 human urothelial carcinoma cells. Immunohistochemical examination showed high expression of ALKBH8 and NOX-1 proteins in high-grade, superficially and deeply invasive carcinomas (pT(1) and >pT(2)) as well as in carcinoma in situ, but not in low-grade and noninvasive phenotypes (pT(a)). These findings indicate an essential role for ALKBH8 in urothelial carcinoma cell survival mediated by NOX-1-dependent ROS signals, further suggesting new therapeutic strategies in human bladder cancer by inducing JNK/p38/gammaH2AX-mediated cell death by silencing of ALKBH8. PMID- 19293183 TI - Thyroid transcription factor-1 inhibits transforming growth factor-beta-mediated epithelial-to-mesenchymal transition in lung adenocarcinoma cells. AB - Thyroid transcription factor-1 (TTF-1) is expressed in lung cancer, but its functional roles remain unexplored. TTF-1 gene amplification has been discovered in a part of lung adenocarcinomas, and its action as a lineage-specific oncogene is highlighted. Epithelial-to-mesenchymal transition (EMT) is a crucial event for cancer cells to acquire invasive and metastatic phenotypes and can be elicited by transforming growth factor-beta (TGF-beta). Mesenchymal-to-epithelial transition (MET) is the inverse process of EMT; however, signals that induce MET are largely unknown. Here, we report a novel functional aspect of TTF-1 that inhibits TGF beta-mediated EMT and restores epithelial phenotype in lung adenocarcinoma cells. This effect was accompanied by down-regulation of TGF-beta target genes, including presumed regulators of EMT, such as Snail and Slug. Moreover, silencing of TTF-1 enhanced TGF-beta-mediated EMT. Thus, TTF-1 can exert a tumor suppressive effect with abrogation of cellular response to TGF-beta and attenuated invasive capacity. We further revealed that TTF-1 down-regulates TGF beta2 production in A549 cells and that TGF-beta conversely decreases endogenous TTF-1 expression, suggesting that enhancement of autocrine TGF-beta signaling accelerates the decrease of TTF-1 expression and vice versa. These findings delineate potential links between TTF-1 and TGF-beta signaling in lung cancer progression through regulation of EMT and MET and suggest that modulation of TTF 1 expression can be a novel therapeutic strategy for treatment of lung adenocarcinoma. PMID- 19293184 TI - Quantitative optical spectroscopy: a robust tool for direct measurement of breast cancer vascular oxygenation and total hemoglobin content in vivo. AB - We propose the use of a robust, biopsy needle-based, fiber-optic tool for routine clinical quantification of tumor oxygenation at the time of diagnostic biopsy for breast cancer. The purpose of this study was to show diffuse reflectance spectroscopy as a quantitative tool to measure oxygenation levels in the vascular compartment of breast cancers in vivo via an optical biopsy technique. Thirty five patients undergoing surgical treatment for breast cancer were recruited for the study at Duke University Medical Center. Diffuse reflectance spectroscopy was performed on the tumors in situ before surgical resection, followed by needle core biopsy of the optically measured tissue. Hemoglobin saturation and total hemoglobin content were quantified from 76 optical spectra-tissue biopsy pairs, consisting of 20 malignant, 23 benign, and 33 adipose tissues. Hemoglobin saturation in malignant tissues was significantly lower than nonmalignant tissues (P<0.002) and was negatively correlated with tumor size and pathologic tumor category (P<0.05). Hemoglobin saturation was positively correlated with total hemoglobin content in malignant tissues (P<0.02). HER2/neu-amplified tumors exhibited significantly higher total hemoglobin content (P<0.05) and significantly higher hemoglobin saturation (P<0.02), which is consistent with a model of increased angiogenesis and tumor perfusion promoted by HER2/neu amplification. Diffuse reflectance spectroscopy could aid in prognosis and prediction in breast cancer via quantitative assessment of tumor physiology at the time of diagnostic biopsy. PMID- 19293185 TI - The EGF/CSF-1 paracrine invasion loop can be triggered by heregulin beta1 and CXCL12. AB - An important step in the process of metastasis from the primary tumor is invasive spread into the surrounding stroma. Using an in vivo invasion assay, we have previously shown that imposed gradients of epidermal growth factor (EGF) or colony-stimulating factor-1 (CSF-1) can induce invasion through an EGF/CSF-1 paracrine loop between cancer cells and macrophages. We now report that invasion induced by other ligands also relies on this EGF/CSF-1 paracrine invasive loop. Using an in vivo invasion assay, we show that MTLn3 breast cancer cells overexpressing ErbB3 exhibit enhanced invasion compared with control MTLn3 cells in response to the ErbB3 ligand HRG-beta1. The invasive response of both MTLn3 ErbB3 and transgenic MMTV-Neu tumors to HRG-beta1 is inhibited by blocking EGF receptor, CSF-1 receptor, or macrophage function, indicating that invasiveness to HRG-beta1 is dependent on the EGF/CSF-1 paracrine loop. Furthermore, we show that CXCL12 also triggers in vivo invasion of transgenic MMTV-PyMT tumors in an EGF/CSF-1-dependent manner. Although the invasion induced by HRG-beta1 or CXCL12 is dependent on the EGF/CSF-1 paracrine loop, invasion induced by EGF is not dependent on HRG-beta1 or CXCL12 signaling, showing an asymmetrical relationship between different ligand/receptor systems in driving invasion. Our results identify a stromal/tumor interaction that acts as an engine underlying invasion induced by multiple ligands. PMID- 19293186 TI - Tumor-infiltrating regulatory dendritic cells inhibit CD8+ T cell function via L arginine metabolism. AB - Dendritic cells (DC) have a critical effect on the outcome of adaptive immune responses against growing tumors. Whereas it is generally assumed that the presence of phenotypically mature DCs should promote protective antitumor immunity, evidence to the contrary does exist. We describe here a novel mechanism by which tumor-infiltrating dendritic cells (TIDC) actively contribute to the suppression of protective CD8(+) T-cell-based antitumor immunity. Using the BALB/NeuT model of spontaneously arising mammary carcinoma, we found that canonical MHC II(+)/CD11b(+)/CD11c(high) TIDCs act as regulatory DCs to suppress CD8(+) T-cell function, resulting in diminished T-cell-based antitumor immunity in vivo. Stimulation of naive T cells with regulatory TIDCs resulted in an altered cell fate program characterized by minimal T-cell expansion, impaired IFNgamma production, and anergy. Suppression by regulatory TIDCs overcame stimulatory signals provided by standard DCs, occurred in the absence of cognate interactions with T cells, and was mediated primarily by arginase metabolism of l arginine. Immunosuppressive TIDCs were found in every murine tumor type examined and were phenotypically distinct from tumor-infiltrating CD11c(int low)/CD11b(+)/Gr-1(+) myeloid-derived suppressor cells. Thus, within the tumor microenvironment, MHC II(+) TIDCs can function as potent suppressors of CD8(+) T cell immunity. PMID- 19293187 TI - PME-1 protects extracellular signal-regulated kinase pathway activity from protein phosphatase 2A-mediated inactivation in human malignant glioma. AB - Extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase pathway activity is regulated by the antagonist function of activating kinases and inactivating protein phosphatases. Sustained ERK pathway activity is commonly observed in human malignancies; however, the mechanisms by which the pathway is protected from phosphatase-mediated inactivation in the tumor tissue remain obscure. Here, we show that methylesterase PME-1-mediated inhibition of the protein phosphatase 2A promotes basal ERK pathway activity and is required for efficient growth factor response. Mechanistically, PME-1 is shown to support ERK pathway signaling upstream of Raf, but downstream of growth factor receptors and protein kinase C. In malignant gliomas, PME-1 expression levels correlate with both ERK activity and cell proliferation in vivo. Moreover, PME-1 expression significantly correlates with disease progression in human astrocytic gliomas (n=222). Together, these observations identify PME-1 expression as one mechanism by which ERK pathway activity is maintained in cancer cells and suggest an important functional role for PME-1 in the disease progression of human astrocytic gliomas. PMID- 19293188 TI - Spleen tyrosine kinase functions as a tumor suppressor in melanoma cells by inducing senescence-like growth arrest. AB - Loss of tumor-suppressive pathways that control cellular senescence is a crucial step in malignant transformation. Spleen tyrosine kinase (Syk) is a cytoplasmic tyrosine kinase that has been recently implicated in tumor suppression of melanoma, a deadly skin cancer derived from pigment-producing melanocytes. However, the mechanism by which Syk suppresses melanoma growth remains unclear. Here, we report that reexpression of Syk in melanoma cells induces a p53 dependent expression of the cyclin-dependent kinase (cdk) inhibitor p21 and a senescence program. We first observed that Syk expression is lost in a subset of melanoma cell lines, primarily by DNA methylation-mediated gene silencing and restored after treatment with the demethylating agent 5-aza-2-deoxycytidine. We analyzed the significance of epigenetic inactivation of Syk and found that reintroduction of Syk in melanoma cells dramatically reduces clonogenic survival and three-dimensional tumor spheroid growth and invasion. Remarkably, melanoma cells reexpressing Syk display hallmarks of senescent cells, including reduction of proliferative activity and DNA synthesis, large and flattened morphology, senescence-associated beta-galactosidase activity, and heterochromatic foci. This phenotype is accompanied by hypophosphorylated retinoblastoma protein (Rb) and accumulation of p21, which depends on functional p53. Our results highlight a new role for Syk tyrosine kinase in regulating cellular senescence and identify Syk mediated senescence as a novel tumor suppressor pathway the inactivation of which may contribute to melanoma tumorigenicity. PMID- 19293189 TI - Inhibition of the sodium potassium adenosine triphosphatase pump sensitizes cancer cells to anoikis and prevents distant tumor formation. AB - Normal epithelial cells undergo apoptosis upon detachment from the extracellular matrix, a process termed "anoikis." However, malignant epithelial cells with metastatic potential resist anoikis and can survive in an anchorage-independent fashion. Molecules that sensitize resistant cells to anoikis will be useful chemical probes to understand this pathway. To identify novel anoikis sensitizers in anoikis-resistant PPC-1 prostate adenocarcinoma cells, a library of 2,000 off patent drugs and natural products was screened for their ability to preferentially induce cell death in suspension over adherent culture conditions. This screen identified five members of the family of cardiac glycosides as anoikis sensitizers, including ouabain, peruvoside, digoxin, digitoxin, and strophanthidin. We conducted further studies with ouabain to discern the mechanism of cardiac glycoside-induced anoikis sensitization. Ouabain initiated anoikis through the mitochondrial pathway of caspase activation. In addition, ouabain sensitized cells to anoikis by inhibiting its known target, the Na(+)/K(+) ATPase pump, and inducing hypoosmotic stress. Resistance to anoikis permits cancer cells to survive in the circulation and facilitates their metastasis to distant organs, so we tested the effects of Na(+)/K(+) ATPase inhibition on distant tumor formation in mouse models. In these mouse models, ouabain inhibited tumor metastases but did not alter the growth of subcutaneous tumors. Thus, we have identified a novel mechanism to sensitize resistant cells to anoikis and decrease tumor metastasis. These results suggest a potential mechanism for the observed clinical reduction in metastasis and relapse in breast cancer patients who have undergone treatments with cardiac glycosides. PMID- 19293190 TI - Chemokine expression in melanoma metastases associated with CD8+ T-cell recruitment. AB - Despite the frequent detection of circulating tumor antigen-specific T cells, either spontaneously or following active immunization or adoptive transfer, immune-mediated cancer regression occurs only in the minority of patients. One theoretical rate-limiting step is whether effector T cells successfully migrate into metastatic tumor sites. Affymetrix gene expression profiling done on a series of metastatic melanoma biopsies revealed a major segregation of samples based on the presence or absence of T-cell-associated transcripts. The presence of lymphocytes correlated with the expression of defined chemokine genes. A subset of six chemokines (CCL2, CCL3, CCL4, CCL5, CXCL9, and CXCL10) was confirmed by protein array and/or quantitative reverse transcription-PCR to be preferentially expressed in tumors that contained T cells. Corresponding chemokine receptors were found to be up-regulated on human CD8(+) effector T cells, and transwell migration assays confirmed the ability of each of these chemokines to promote migration of CD8(+) effector cells in vitro. Screening by chemokine protein array identified a subset of melanoma cell lines that produced a similar broad array of chemokines. These melanoma cells more effectively recruited human CD8(+) effector T cells when implanted as xenografts in nonobese diabetic/severe combined immunodeficient mice in vivo. Chemokine blockade with specific antibodies inhibited migration of CD8(+) T cells. Our results suggest that lack of critical chemokines in a subset of melanoma metastases may limit the migration of activated T cells, which in turn could limit the effectiveness of antitumor immunity. PMID- 19293191 TI - The membrane mucin Muc4 inhibits apoptosis induced by multiple insults via ErbB2 dependent and ErbB2-independent mechanisms. AB - The aberrant expression of membrane mucins such as Muc1 and Muc4 by tumor cells has been shown to engage signaling pathways that promote cellular properties associated with tumor progression. Our previous studies have shown that Muc4 interacts with and potentiates signaling by the ErbB2 (HER2) receptor tyrosine kinase through an epidermal growth factor-like domain in its extracellular region. Here, we show that expression of Muc4 in human A375 melanoma cells and MCF7 breast cancer cells confers resistance to apoptosis induced by a variety of stimuli, including chemotherapeutic agents, the absence of serum factors, and the loss of cellular adhesion. Mapping experiments revealed that the O-glycosylation and cytosolic domains of Muc4 are dispensable for its antiapoptotic activity, and are also dispensable for the potentiation of signaling by ErbB2. Knockdown of endogenous Muc4 in JIMT-1 breast cancer cells sensitizes cells to apoptotic stimuli, and this can be rescued by Muc4 forms lacking the O-glycosylation or cytosolic domains. Surprisingly, however, the molecular mechanisms underlying Muc4 antiapoptotic activity vary among cell lines. Although Muc4 in JIMT-1 cells engages ErbB2 to promote cell survival, its antiapoptotic mechanism in MCF7 and A375 cells seems to be independent of ErbB2. However, Muc4 expression in all cell lines culminates in the phosphorylation and inactivation of the proapoptotic protein Bad and the elevation of the prosurvival protein Bcl-xL. Our observations suggest that tumor cells can exploit the versatile antiapoptotic activities of Muc4 to acquire resistance to therapeutic agents, and augment cell survival after the loss of adhesion and microenvironment-derived survival factors. PMID- 19293192 TI - Cathepsin G recruits osteoclast precursors via proteolytic activation of protease activated receptor-1. AB - Metastatic breast cancer shows extreme tropism for the bone microenvironment, leading to the establishment of osteolytic metastases. Perpetuation of tumor induced osteolysis requires a continuous supply of osteoclast precursors migrating into the bone microenvironment that can subsequently differentiate into mature osteoclasts and resorb bone. Thus, identification and subsequent targeting of chemoattractants of osteoclast precursors that are up-regulated at the tumor bone interface represents a potential avenue to interrupt osteolysis. We report that cathepsin G, a serine protease, plays a vital role in the bone microenvironment by modulating tumor-stromal interaction in a manner that favors tumor establishment and regulates chemotaxis of monocytes, a subset of which has the potential to differentiate into osteoclasts. Our data show that cathepsin G induced chemotaxis of monocytes is mediated by proteolytic activation of protease activated receptor-1 (PAR-1). Attenuation of PAR-1 activation abrogates cathepsin G-mediated induction of monocyte chemotaxis. We also show that in vivo inhibition of cathepsin G reduces the number of CD11b(+) osteoclast precursors and mature osteoclasts at the tumor-bone interface. Together, these data suggest that therapeutic targeting of both PAR-1 signaling in osteoclast precursors as well as cathepsin G at the tumor-bone interface has the potential to reduce osteolysis by inhibiting the recruitment, differentiation, and activation of osteoclast precursors. PMID- 19293193 TI - Central and peripheral vagal nerve involvement in atrial fibrillation. PMID- 19293194 TI - Aspirin following PCI: too much of a good thing? PMID- 19293195 TI - Risk factors for myocardial infarction in women and men. PMID- 19293196 TI - Impact of intravenous {beta}-lactam/macrolide versus {beta}-lactam monotherapy on mortality in hospitalized patients with community-acquired pneumonia. AB - OBJECTIVES: Guidelines recommend dual-therapy consisting of a beta lactam/macrolide (BLM) for hospitalized patients with community-acquired pneumonia. Nevertheless, the superiority over beta-lactam-monotherapy (BL) remains unproven. METHODS: Analyses from an observational study initiated by the German competence network CAPNETZ were performed. RESULTS: One thousand eight hundred and fifty-four patients were treated with either BL (49.0%) or BLM (51.0%). BLM therapy was associated with lower adjusted 14 day mortality [odds ratio (OR) 0.53; 95% confidence interval (CI): 0.30-0.94]. CRB65, neoplastic disease, age and nursing home residency were confirmed as independent predictors of death. Adjusted 14 day mortality risk was clearly reduced in patients with CRB65 = 2 (n = 411; OR 0.35; CI: 0.12-0.99) and CRB65 > or = 2 (n = 519; OR 0.42; CI: 0.18-0.997). However, this could not be shown for adjusted 30 day mortality. Patients with CRB65 < or = 1 showed low mortality (2.1%) without the influence of BLM. BLM therapy was associated with lower adjusted risk of treatment failure at 14 days (n = 1854; OR 0.65; CI: 0.47-0.89) and 30 days (OR 0.69; CI: 0.51-0.94) as well as in the subgroup of patients with CRB65 = 2 and CRB65 > or = 2. CONCLUSIONS: This study suggests the superiority of BLM therapy in patients with CRB65 risk classes of 2 or higher on 14 day mortality. BLM therapy was also associated with lower risk of treatment failure. PMID- 19293197 TI - Polymorphisms of the beta2 adrenoreceptor gene in chronic obstructive pulmonary disease. AB - BACKGROUND: The beta2-adrenergic receptors are cell surface receptors playing a central role in the pharmacological targeting asthma and chronic obstructive pulmonary disease [COPD]. Recent studies suggest that patients who are homozygous for one of the two important polymorphisms of the beta2-adrenergic receptor [ADRB2] gene at codon 16 (arginine to glycine) and 27 (glutamine to glutamate) may have a reduced response to ss2-agonists. Since smoking patients who are Gly16 homozygotes have an increased risk of airway obstruction we hypothesized that beta2-adrenoreceptor gene polymorphisms may be also a cofounder for COPD development and disease severity. METHODS: We investigated 190 COPD patients and 172 healthy volunteers in a case-control study. DNA was isolated from whole blood and beta2-AR gene polymorphisms Arg/Gly16 and Gln/Glu27 were determined using allele-specific polymerase chain reaction [PCR]. RESULTS: In COPD patients with Gly/Gly16 was found more frequently than in healthy smokers [29.47% COPD versus 18.18% controls, p = 0.026]. All other gene polymorphisms of the ADRB2 gene at codon 16 were equally distributed between groups. ss2-adrenoreceptor gene polymorphisms were neither a cofounder for COPD exacerbations [>or= 3 hospitalizations within the last 3 years] nor for disease severity [FEV1 0.8, MAF>0.05] and were genotyped using TaqMan genotyping technologies. Quantitative genetic association was assessed using regression modelling to correct for covariates. RESULTS: in patients with AATD, carriers of the G allele of rs678815 [MMP3] had lower gas transfer [KCO] [P = 0.025, B =-7.766] than the homozygous wild type, while carriers of the T allele of rs470358 [MMP1] had higher KCO [P = 0.025, B = 6.130]. CONCLUSIONS: variations in MMP1 and MMP3 are associated with gas transfer in AATD, supporting a previous family study showing linkage of KCO to this gene region. Replication of these preliminary data is now required particularly if MMP inhibitors are to be considered as a therapeutic option. PMID- 19293201 TI - Smoking and depression: Is smoking cessation effective? AB - Patients with depression tend to have a higher rate of smoking and an increased severity of nicotine dependence. It seems that common genetic and environmental factors may influence the bimodal relationship between smoking and depression. Nicotine has some positive reinforcing/rewarding effects that may affect people with mood disorders and smoking is also used as a ;relief medication' in order to ameliorate symptoms of depression, as it is already known that nicotine, which is the major psychoactive ingredient in tobacco, may act as an antidepressant. To date, the data regarding the difficulty in smoking cessation in depressed smokers are conflicting. Low confidence and self-esteem among this group of smokers are significant predictors of failure during smoking cessation attempt. In the most recently published guidelines for smokers with psychiatric comorbidities the suggestion is for combination treatment (counseling and pharmaceutical treatment) and prolongation of a therapeutic approach. PMID- 19293202 TI - From large clinical trials to management of COPD in the real world. AB - Large clinical trials in chronic obstructive pulmonary disease [COPD] are analyzed and discussed. Unfortunately, all of them have failed to reach their primary endpoint, which has mainly been the effect on the rate of decline in mean FEV(1) [forced expiratory volume in 1 second]. Nonetheless, almost all trials have demonstrated benefits in important outcomes such as exacerbation frequency, symptoms, quality of life and other measures of health status, which are arguably more meaningful to individual patients than FEV( 1) per se. PMID- 19293203 TI - Antiprotozoal drugs in the treatment of respiratory allergy: a side that should be explored. PMID- 19293204 TI - Utilization of imaging in pregnant patients: 10-year review of 5270 examinations in 3285 patients--1997-2006. AB - PURPOSE: To document the utilization of radiologic imaging in pregnant patients at one academic institution during a 10-year period (1997-2006). MATERIALS AND METHODS: The study was approved by the hospital institutional review board and was compliant with HIPAA. Informed consent was waived. At the authors' institution, pregnant patients exposed to radiation during imaging are recorded in a database compiled by the medical physics department. The authors retrospectively reviewed this database to document the number of patients, number of each type of imaging examination, date of the examination, and the estimated radiation dose to the fetus from 1997 to 2006. The authors searched the institution's medical records to obtain the total number of deliveries by year as a control for the total pregnant patient population. RESULTS: During the 10-year period, 5270 examinations were performed in 3285 pregnant patients (mean age, 28 years). The number of patients and examinations increased from 237 patients undergoing 331 studies in 1997 to 449 patients undergoing 732 examinations in 2006, an increase of 89% in patients and 121% in examinations. The total number of pregnant patients measured by deliveries increased 7%--from 8661 in 1997 to 9264 in 2006. Utilization rates (examinations per 1000 deliveries) of all radiologic examinations increased 107% from 1997 to 2006. The number of conventional radiographic examinations increased by an average of 7% per year, nuclear medicine examinations by 12% per year, and computed tomographic (CT) examinations by 25% per year. The average estimated fetal radiation exposure per examination was 0.43 mGy (range, 0.01-22.5 mGy) for conventional radiography, 4.3 mGy (range, 0.01-43.9 mGy) for CT, and 0.40 mGy (range, 0.01-7.7 mGy) for nuclear medicine examinations. CONCLUSION: For the comparison of 1997 to 2006, the radiologic utilization rate in pregnant patients increased by 107% from 1997 to 2006. The greatest increase was in CT. PMID- 19293205 TI - Alexander technique trial. A trial subject's perspective. PMID- 19293206 TI - Orthopaedic attendance also worse with choose and book. PMID- 19293207 TI - Mobile phones in hospital. Resurgence in memorial postmortem photography? PMID- 19293208 TI - Immediate breast reconstruction should be challenged. PMID- 19293209 TI - Measuring up the NHS. The story of outperforming expectations continues. PMID- 19293210 TI - Coeliac disease. Don't forget increased risk of fetal growth restriction. PMID- 19293211 TI - Coeliac disease. Non-invasive diagnosis needed. PMID- 19293212 TI - Nurse delivered endoscopy. Opportunity costs of endoscopy. PMID- 19293213 TI - On criticising Israel. A thick skin may not provide protection. PMID- 19293214 TI - On criticising Israel. In praise of pachydermatous truth. PMID- 19293215 TI - On criticising Israel. Stop de-legitimising Israel and politicising your journal. PMID- 19293216 TI - On criticising Israel. Response from author of original paper. Comparison of the number of Palestinians killed by the Israeli Defence Force (IDF) with those killed in 9/11. PMID- 19293218 TI - Study authors respond to editorial. Nurse delivered endoscopy. PMID- 19293219 TI - Inequality can lead to poor mental health and depression, report says. PMID- 19293220 TI - Dutch public health experts refute claims that human papillomavirus vaccination has health risks. PMID- 19293222 TI - Target cheap drinking as we did passive smoking, says chief medical officer. PMID- 19293223 TI - Will doctor rating sites improve standards of care? Yes. PMID- 19293224 TI - Will doctor rating sites improve the quality of care? No. PMID- 19293225 TI - Screening is effective in early detection of ovarian cancer. PMID- 19293227 TI - Latest figures show reduction in hospital admissions for stab wounds. PMID- 19293228 TI - High court rejects challenge to NICE guidelines on chronic fatigue syndrome. PMID- 19293229 TI - Strengthen clinical trials driven by investigators, Europe research councils say. PMID- 19293230 TI - Increase in reports of medical errors suggest NHS staff are more confident in reporting problems. PMID- 19293232 TI - Compensation of loss of protein function in microsatellite-unstable colon cancer cells (HCT116): a gene-dependent effect on the cell surface glycan profile. AB - Tumors that display a high level of microsatellite instability (MSI-H) accumulate somatic frameshift mutations in several genes. The compensation of this loss of function by transfection represents a suitable approach to tie respective gene deficiency to alterations in cellular characteristics. In view of the emerging significance of cell surface glycans as biochemical signals for presentation/activity of various receptors/integrins and for susceptibility to adhesion/growth-regulatory tissue lectins, we examined the glycophenotype in the MSI-H colon cancer cell line HCT116 for activin type 2 receptor (ACVR2), absent in melanoma 2 (AIM2), and transforming growth factor beta-type 2 receptor (TGFBR2) known to be associated with MSI colorectal carcinogenesis. A panel of probes specific for functional carbohydrate epitopes including human lectins was used to trace changes in cell surface levels, thereby initiating glycan analysis related to MSI. In particular, the presence of core substitutions and branching in N-glycans, the sialylation status of N- and O-glycans, and the presence of Le(a/x)-epitopes were profiled. Transient transfection affected the glycophenotype, depending on the nature of the gene and the probe. The TGFBR2 presence reduced binding of probes specific for a core substitution and increased branch length in N-glycosylation, even reaching a P-value of 0.0016. ACVR2/AIM2 influenced core 1 mucin-type O-glycosylation differentially, upregulation by ACVR2, and downregulation by AIM2. These alterations of cell surface glycosylation by gene products that are not directly associated with the machinery for glycan generation direct attention to pursue analysis of glycosylation in MSI tumor cells on the level of target glycoproteins and open the way for functional studies. PMID- 19293233 TI - Involvement of chondroitin sulfate E in the liver tumor focal formation of murine osteosarcoma cells. AB - Cell surface heparan sulfate plays a critical role in regulating the metastatic behavior of tumor cells, whereas the role of chondroitin sulfate/dermatan sulfate (CS/DS) has been little understood in this context. Here, we characterized CS/DS chains from the murine osteosarcoma cell line LM8G7, which forms tumor nodules in liver. Structural analysis of the CS/DS chains showed a higher proportion of GlcUA beta 1-3GalNAc(4,6-O-disulfate) (E-units) in LM8G7 (12%) than in its parental cell line LM8 (6%), which rarely forms tumors in the liver. Immunostaining with GD3G7, an antibody specific to E-units, confirmed the higher expression of the epitope in LM8G7 than LM8 cells. The tumor focal formation of LM8G7 cells in the liver in mice was effectively inhibited by the preadministration of CS-E (rich in E-unit) or the preincubation of the antibody GD3G7 with the tumor cells. CS-E or GD3G7 inhibited the adhesion of LM8G7 cells to a laminin-coated plate in vitro. In addition, the invasive ability of LM8G7 cells in vitro was also reduced by the addition of CS-E or the antibody. Further, CS-E or the antibody inhibited the proliferation of LM8G7 cells dose dependently. The binding of LM8G7 cells to VEGF in vitro was also significantly reduced by CS E and GD3G7. Thus, the present study reveals the significance of highly sulfated CS/DS structures in the liver colonization of osteosarcoma cells and also provides a framework for the development of GAG-based anticancer molecules. PMID- 19293235 TI - Distinct phenotypic and functional features of CADASIL mutations in the Notch3 ligand binding domain. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an autosomal dominant small-vessel disease of the brain caused by mutations in the NOTCH3 receptor. The highly stereotyped nature of the mutations, which alter the number of cysteine residues within the epidermal growth factor-like repeats (EGFR), predicts that all mutations share common mechanisms. Prior in vitro assays and genetic studies in the mouse support the hypothesis that common mutations do not compromise canonical Notch3 function but instead convey a non-physiological and deleterious activity to the receptor through the unpaired cysteine residue. Intriguingly, in vitro studies predict that mutations located in the Delta/Serrate/LAG-2 ligand binding domain-(EGFR10 11) may result in a loss of Notch3 receptor function. However, the in vivo relevance and functional significance of this with respect to the pathogenic mechanisms and clinical expression of the disease remain largely unexplored. To ascertain, in vivo, the functional significance of EGFR10-11 mutations, we generated transgenic mice with one representative mutation (C428S) in EGFR10 of Notch3. These mice, like those with a common R90C mutation, developed characteristic arterial accumulation of Notch3 protein and granular osmiophilic material upon aging. By introducing the mutant C428S transgene into a Notch3 null background, we found that, unlike the R90C mutant protein, the C428S mutant protein has lost wild-type Notch3 activity and exhibited mild dominant-negative activity in three different biological settings. From a large prospectively recruited cohort of 176 CADASIL patients, we identified 10 patients, from five distinct pedigrees carrying a mutation in EGFR10 or 11. These mutations were associated with significantly higher Mini-Mental State Examination and Mattis Dementia Rating Scale scores (P < 0.05), when compared with common mutations. Additionally, we found a strong effect of this genotype on the burden of white matter hyperintensities (P < 0.01). Collectively, these results highlight distinctive functional and phenotypic features of EGFR10-11 mutations relative to the common CADASIL mutations. Our findings are compatible with the hypothesis that EGFR10-11 mutations cause the disease through the same gain of novel function as the common mutations, and lead us to propose that reduced Notch3 signalling acts as a modifier of the CADASIL phenotype. PMID- 19293236 TI - Short-term memory binding deficits in Alzheimer's disease. AB - Alzheimer's disease impairs long term memories for related events (e.g. faces with names) more than for single events (e.g. list of faces or names). Whether or not this associative or 'binding' deficit is also found in short-term memory has not yet been explored. In two experiments we investigated binding deficits in verbal short-term memory in Alzheimer's disease. Experiment 1: 23 patients with Alzheimer's disease and 23 age and education matched healthy elderly were recruited. Participants studied visual arrays of objects (six for healthy elderly and four for Alzheimer's disease patients), colours (six for healthy elderly and four for Alzheimer's disease patients), unbound objects and colours (three for healthy elderly and two for Alzheimer's disease patients in each of the two categories), or objects bound with colours (three for healthy elderly and two for Alzheimer's disease patients). They were then asked to recall the items verbally. The memory of patients with Alzheimer's disease for objects bound with colours was significantly worse than for single or unbound features whereas healthy elderly's memory for bound and unbound features did not differ. Experiment 2: 21 Alzheimer's disease patients and 20 matched healthy elderly were recruited. Memory load was increased for the healthy elderly group to eight items in the conditions assessing memory for single or unbound features and to four items in the condition assessing memory for the binding of these features. For Alzheimer's disease patients the task remained the same. This manipulation permitted the performance to be equated across groups in the conditions assessing memory for single or unbound features. The impairment in Alzheimer's disease patients in recalling bound objects reported in Experiment 1 was replicated. The binding cost was greater than that observed in the healthy elderly group, who did not differ in their performance for bound and unbound features. Alzheimer's disease grossly impairs the mechanisms responsible for holding integrated objects in verbal short term memory. PMID- 19293237 TI - Mitochondrial changes within axons in multiple sclerosis. AB - Multiple sclerosis is the most common cause of non-traumatic neurological impairment in young adults. An energy deficient state has been implicated in the degeneration of axons, the pathological correlate of disease progression, in multiple sclerosis. Mitochondria are the most efficient producers of energy and play an important role in calcium homeostasis. We analysed the density and function of mitochondria using immunohistochemistry and histochemistry, respectively, in chronic active and inactive lesions in progressive multiple sclerosis. As shown before in acute pattern III and Balo's lesions, the mitochondrial respiratory chain complex IV activity is reduced despite the presence of mitochondria in demyelinated axons with amyloid precursor protein accumulation, which are predominantly located at the active edge of chronic active lesions. Furthermore, the strong non-phosphorylated neurofilament (SMI32) reactivity was associated with a significant reduction in complex IV activity and mitochondria within demyelinated axons. The complex IV defect associated with axonal injury may be mediated by soluble products of innate immunity, as suggested by an inverse correlation between complex IV activity and macrophage/microglial density in chronic lesions. However, in inactive areas of chronic multiple sclerosis lesions the mitochondrial respiratory chain complex IV activity and mitochondrial mass, judged by porin immunoreactivity, are increased within approximately half of large (>2.5 microm diameter) chronically demyelinated axons compared with large myelinated axons in the brain and spinal cord. The axon-specific mitochondrial docking protein (syntaphilin) and phosphorylated neurofilament-H were increased in chronic lesions. The lack of complex IV activity in a proportion of Na(+)/K(+) ATPase alpha-1 positive demyelinated axons supports axonal dysfunction as a contributor to neurological impairment and disease progression. Furthermore, in vitro studies show that inhibition of complex IV augments glutamate-mediated axonal injury (amyloid precursor protein and SMI32 reactivity). Our findings have important implications for both axonal degeneration and dysfunction during the progressive stage of multiple sclerosis. PMID- 19293238 TI - Too much of a good thing: increased grey matter in boys with conduct problems and callous-unemotional traits. PMID- 19293239 TI - Clinical field-strength MRI of amyloid plaques induced by low-level cholesterol feeding in rabbits. AB - Two significant barriers have limited the development of effective treatment of Alzheimer's disease. First, for many cases the aetiology is unknown and likely multi-factorial. Among these factors, hypercholesterolemia is a known risk predictor and has been linked to the formation of beta-amyloid plaques, a pathological hallmark this disease. Second, standardized diagnostic tools are unable to definitively diagnose this disease prior to death; hence new diagnostic tools are urgently needed. Magnetic resonance imaging (MRI) using high field strength scanners has shown promise for direct visualization of beta-amyloid plaques, allowing in vivo longitudinal tracking of disease progression in mouse models. Here, we present a new rabbit model for studying the relationship between cholesterol and Alzheimer's disease development and new tools for direct visualization of beta-amyloid plaques using clinical field-strength MRI. New Zealand white rabbits were fed either a low-level (0.125-0.25% w/w) cholesterol diet (n = 5) or normal chow (n = 4) for 27 months. High-resolution (66 x 66 x 100 microm(3); scan time = 96 min) ex vivo MRI of brains was performed using a 3 Tesla (T) MR scanner interfaced with customized gradient and radiofrequency coils. Beta-amyloid-42 immunostaining and Prussian blue iron staining were performed on brain sections and MR and histological images were manually registered. MRI revealed distinct signal voids throughout the brains of cholesterol-fed rabbits, whereas minimal voids were seen in control rabbit brains. These voids corresponded directly to small clusters of extracellular beta amyloid-positive plaques, which were consistently identified as iron-loaded (the presumed source of MR contrast). Plaques were typically located in the hippocampus, parahippocampal gyrus, striatum, hypothalamus and thalamus. Quantitative analysis of the number of histologically positive beta-amyloid plaques (P < 0.0001) and MR-positive signal voids (P < 0.05) found in cholesterol fed and control rabbit brains corroborated our qualitative observations. In conclusion, long-term, low-level cholesterol feeding was sufficient to promote the formation of extracellular beta-amyloid plaque formation in rabbits, supporting the integral role of cholesterol in the aetiology of Alzheimer's disease. We also present the first evidence that MRI is capable of detecting iron associated beta-amyloid plaques in a rabbit model of Alzheimer's disease and have advanced the sensitivity of MRI for plaque detection to a new level, allowing clinical field-strength scanners to be employed. We believe extension of these technologies to an in vivo setting in rabbits is feasible and that our results support future work exploring the role of MRI as a leading imaging tool for this debilitating and life-threatening disease. PMID- 19293240 TI - Evaluation of evoked potentials to dyadic tones after cochlear implantation. AB - Auditory evoked potentials are tools widely used to assess auditory cortex functions in clinical context. However, in cochlear implant users, electrophysiological measures are challenging due to implant-created artefacts in the EEG. Here, we used independent component analysis to reduce cochlear implant related artefacts in event-related EEGs of cochlear implant users (n = 12), which allowed detailed spatio-temporal evaluation of auditory evoked potentials by means of dipole source analysis. The present study examined hemispheric asymmetries of auditory evoked potentials to musical sounds in cochlear implant users to evaluate the effect of this type of implantation on neuronal activity. In particular, implant users were presented with two dyadic tonal intervals in an active oddball design and in a passive listening condition. Principally, the results show that independent component analysis is an efficient approach that enables the study of neurophysiological mechanisms of restored auditory function in cochlear implant users. Moreover, our data indicate altered hemispheric asymmetries for dyadic tone processing in implant users compared with listeners with normal hearing (n = 12). We conclude that the evaluation of auditory evoked potentials are of major relevance to understanding auditory cortex function after cochlear implantation and could be of substantial clinical value by indicating the maturation/reorganization of the auditory system after implantation. PMID- 19293241 TI - Dual mechanism of brain injury and novel treatment strategy in maple syrup urine disease. AB - Maple syrup urine disease (MSUD) is an inherited disorder of branched-chain amino acid metabolism presenting with life-threatening cerebral oedema and dysmyelination in affected individuals. Treatment requires life-long dietary restriction and monitoring of branched-chain amino acids to avoid brain injury. Despite careful management, children commonly suffer metabolic decompensation in the context of catabolic stress associated with non-specific illness. The mechanisms underlying this decompensation and brain injury are poorly understood. Using recently developed mouse models of classic and intermediate maple syrup urine disease, we assessed biochemical, behavioural and neuropathological changes that occurred during encephalopathy in these mice. Here, we show that rapid brain leucine accumulation displaces other essential amino acids resulting in neurotransmitter depletion and disruption of normal brain growth and development. A novel approach of administering norleucine to heterozygous mothers of classic maple syrup urine disease pups reduced branched-chain amino acid accumulation in milk as well as blood and brain of these pups to enhance survival. Similarly, norleucine substantially delayed encephalopathy in intermediate maple syrup urine disease mice placed on a high protein diet that mimics the catabolic stress shown to cause encephalopathy in human maple syrup urine disease. Current findings suggest two converging mechanisms of brain injury in maple syrup urine disease including: (i) neurotransmitter deficiencies and growth restriction associated with branched-chain amino acid accumulation and (ii) energy deprivation through Krebs cycle disruption associated with branched-chain ketoacid accumulation. Both classic and intermediate models appear to be useful to study the mechanism of brain injury and potential treatment strategies for maple syrup urine disease. Norleucine should be further tested as a potential treatment to prevent encephalopathy in children with maple syrup urine disease during catabolic stress. PMID- 19293242 TI - On the pathogenesis of collagen VI muscular dystrophies--comment on article of Hicks et al. PMID- 19293244 TI - Incidence and prognosis of > or = 50% symptomatic vertebral or basilar artery stenosis: prospective population-based study. AB - The higher risk of early recurrent stroke after posterior circulation transient ischaemic attack or minor stroke versus after carotid territory events could be due to a greater prevalence of large artery stenosis, but there have been few imaging studies, and the prognostic significance of such stenoses is uncertain. Reliable data are necessary to determine the feasibility of trials of angioplasty and stenting and to inform imaging strategies. In the first-ever population-based study, we determined the prevalence of > or = 50% apparently symptomatic vertebral and basilar stenosis using contrast-enhanced MRA in consecutive patients, irrespective of age, presenting with posterior circulation transient ischaemic attack or minor ischaemic stroke in the Oxford Vascular Study and related this to the 90-day risk of recurrent transient ischaemic attack and stroke. For comparison, we also determined the prevalence of > or = 50% apparently symptomatic carotid stenosis on ultrasound imaging in consecutive patients with carotid territory events. Of 538 consecutive patients, 141/151 (93%) had posterior circulation events and had vertebral and basilar imaging, of whom 37 (26.2%) had > or = 50% vertebral and basilar stenosis, compared with 41 (11.5%) patients with > or = 50% ipsilateral carotid stenosis in 357/387 (92%) patients with carotid events who had carotid imaging (OR = 2.74; 95% CI = 1.67 4.51; P = 0.002). Presence of > or = 50% vertebral and basilar stenosis was unrelated to age, sex or vascular risk factors and, in contrast to > or = 50% carotid stenosis was not associated with evidence of coronary/peripheral atherosclerosis. In patients with posterior circulation events, > or = 50% vertebral and basilar stenosis was associated multiple transient ischaemic attacks at presentation (22% versus 3%; OR = 9.29; 95% CI = 2.31-37.27; P < 0.001) and with a significantly higher 90-day risk of recurrent events (OR = 3.2; 95% CI = 1.4-7.0; P = 0.006), reaching 22% for stroke and 46% for transient ischaemic attack and stroke. The prevalence of > or = 50% vertebral and basilar stenosis in posterior circulation transient ischaemic attack or minor stroke is greater than the prevalence of > or = 50% carotid stenosis in carotid territory events, and is associated with multiple transient ischaemic attacks at presentation and a high early risk of recurrent stroke. Trials of interventional treatment are therefore likely to be feasible, but more data are required on the long-term risk of stroke on best medical treatment. PMID- 19293245 TI - Size matters: increased grey matter in boys with conduct problems and callous unemotional traits. AB - Brain imaging studies of adults with psychopathy have identified structural and functional abnormalities in limbic and prefrontal regions that are involved in emotion recognition, decision-making, morality and empathy. Among children with conduct problems, a small subgroup presents callous-unemotional traits thought to be antecedents of psychopathy. No structural brain imaging study has examined this subgroup of children. The present study used voxel-based morphometry to compare whole brain grey matter volumes and concentrations of boys with elevated levels of callous-unemotional conduct problems and typically developing boys and explored four a priori regions of interest. sMRI scans were collected from 23 boys with elevated levels of callous-unemotional conduct problems (mean age = 11 years 8 months) and 25 typically developing boys (mean age = 11 years 6 months) selected from a community sample of children. Data were analysed using optimized voxel-based morphometry. Study-specific probability maps were created and four a priori regions of interest identified (orbitofrontal, anterior cingulate and anterior insular cortices and amygdala). Both grey matter volume and concentration were examined controlling for cognitive ability and hyperactivity inattention symptoms. Boys with callous-unemotional conduct problems, as compared with typically developing boys, presented increased grey matter concentration in the medial orbitofrontal and anterior cingulate cortices, as well as increased grey matter volume and concentration in the temporal lobes bilaterally. These findings may indicate a delay in cortical maturation in several brain areas implicated in decision making, morality and empathy in boys with callous unemotional conduct problems. PMID- 19293246 TI - Obesity induces functional astrocytic leptin receptors in hypothalamus. AB - The possible role of astrocytes in the regulation of feeding has been overlooked. It is well-established that the endothelial cells constituting the blood-brain barrier transport leptin from blood to brain and that hypothalamic neurons respond to leptin to induce anorexic signaling. However, few studies have addressed the role of astrocytes in either leptin transport or cellular activation. We recently showed that the obese agouti viable yellow mouse has prominent astrocytic expression of the leptin receptor. In this study, we test the hypothesis that diet-induced obesity increases astrocytic leptin receptor expression and function in the hypothalamus. Double-labelling immunohistochemistry and confocal microscopic analysis showed that all astrocytes in the hypothalamus express leptin receptors. In adult obese mice, 2 months after being placed on a high-fat diet, there was a striking increase of leptin receptor (+) astrocytes, most prominent in the dorsomedial hypothalamus and arcuate nucleus. Agouti viable yellow mice with their adult-onset obesity showed similar changes, but the increase of leptin receptor (+) astrocytes was barely seen in ob/ob or db/db mice with their early-onset obesity and defective leptin systems. The marked leptin receptor protein expression in the astrocytes, shown with several antibodies against different receptor epitopes, was supported by RT-PCR detection of leptin receptor-a and -b mRNAs in primary hypothalamic astrocytes. Unexpectedly, the protein expression of GFAP, a marker of astrocytes, was also increased in adult-onset obesity. Real-time confocal imaging showed that leptin caused a robust increase of calcium signalling in primary astrocytes from the hypothalamus, confirming their functionality. The results indicate that metabolic changes in obese mice can rapidly alter leptin receptor expression and astrocytic activity, and that leptin receptor is responsible for leptin-induced calcium signalling in astrocytes. This novel and clinically relevant finding opens new avenues in astrocyte biology. PMID- 19293247 TI - Growth hormone-releasing hormone promotes survival of cardiac myocytes in vitro and protects against ischaemia-reperfusion injury in rat heart. AB - AIMS: The hypothalamic neuropeptide growth hormone-releasing hormone (GHRH) stimulates GH synthesis and release in the pituitary. GHRH also exerts proliferative effects in extrapituitary cells, whereas GHRH antagonists have been shown to suppress cancer cell proliferation. We investigated GHRH effects on cardiac myocyte cell survival and the underlying signalling mechanisms. METHODS AND RESULTS: Reverse transcriptase-polymerase chain reaction analysis showed GHRH receptor (GHRH-R) mRNA in adult rat ventricular myocytes (ARVMs) and in rat heart H9c2 cells. In ARVMs, GHRH prevented cell death and caspase-3 activation induced by serum starvation and by the beta-adrenergic receptor agonist isoproterenol. The GHRH-R antagonist JV-1-36 abolished GHRH survival action under both experimental conditions. GHRH-induced cardiac cell protection required extracellular signal-regulated kinase (ERK)1/2 and phosphoinositide-3 kinase (PI3K)/Akt activation and adenylyl cyclase/cAMP/protein kinase A signalling. Isoproterenol strongly upregulated the mRNA and protein of the pro-apoptotic inducible cAMP early repressor, whereas GHRH completely blocked this effect. Similar to ARVMs, in H9c2 cardiac cells, GHRH inhibited serum starvation- and isoproterenol-induced cell death and apoptosis through the same signalling pathways. Finally, GHRH improved left ventricular recovery during reperfusion and reduced infarct size in Langendorff-perfused rat hearts, subjected to ischaemia reperfusion (I/R) injury. These effects involved PI3K/Akt signalling and were inhibited by JV-1-36. CONCLUSION: Our findings suggest that GHRH promotes cardiac myocyte survival through multiple signalling mechanisms and protects against I/R injury in isolated rat heart, indicating a novel cardioprotective role of this hormone. PMID- 19293248 TI - Different extent of cardiac malfunction and resistance to oxidative stress in heterozygous and homozygous manganese-dependent superoxide dismutase-mutant mice. AB - AIMS: The mitochondrially expressed manganese-dependent superoxide dismutase (MnSOD, SOD2) is an essential antioxidative enzyme that is necessary for normal heart function. In this study, we investigated the heart function of mice that were exposed to increased oxidative stress for time periods of up to 6 months due to decreased MnSOD activity caused by heterozygous deletion of the MnSOD gene. METHODS AND RESULTS: We generated a mouse strain in which the gene encoding MnSOD was exchanged against a cassette containing the SOD cDNA under the control of the tetracycline response element. After breeding with mice carrying the tetracycline receptor, compound mice express MnSOD depending on the presence of tetracycline. Without tetracycline receptor the MnSOD gene is fully inactivated, and animals show an MnSOD-deficient phenotype. Using echocardiographic recordings, we found an impairment of left ventricular functions: MnSOD+/- mice displayed a decrease in fraction shortening and ejection fraction and an increase in left ventricular internal diameter in systole. Furthermore, MnSOD+/- mice developed heart hypertrophy with accompanying fibrosis and necrosis revealed by immunhistochemical analysis. Although we did not find an increase in apoptosis in MnSOD+/- hearts under normal conditions, we observed an increase of the number of apoptotic cells and vascular senescence after treatment with doxorubicin. CONCLUSION: Our study demonstrates that lifelong reduction of MnSOD activity has a negative effect on normal heart function. This animal model presents a valuable tool to investigate the mechanism of heart pathology reported in patients bearing different polymorphic variants of the MnSOD gene and to develop new therapeutic strategies through manipulation of the antioxidative defence system. PMID- 19293249 TI - Identification of multiple dmrt1s in catfish: localization, dimorphic expression pattern, changes during testicular cycle and after methyltestosterone treatment. AB - The double sex and mab-3 related (DM) transcription factor 1 (dmrt1) plays an important role in testicular differentiation. Here, we report cloning of multiple dmrt1s, a full-length and two alternative spliced forms from adult catfish (Clarias gariepinus) testis, which encode predicted proteins of 287 (dmrt1a), 253 (dmrt1b) and 233 (dmrt1c) amino acid residues respectively. Interestingly, dmrt1c lacks the majority of the DM domain. Multiple dmrt1s (dmrt1a and dmrt1c) were obtained from Clarias batrachus as well. Tissue distribution (transcript and protein) of catfish dmrt1 revealed exclusive expression in testis. Semi quantitative RT-PCR revealed the presence of multiple dmrt1s with high levels of dmrt1a in adult testis but not in ovary. Real-time RT-PCR analysis during testicular cycle showed higher levels of dmrt1 transcripts in preparatory and pre spawning when compared with spawning and post-spawning phases. Immunocytochemical and immunofluorescence localization revealed the presence of catfish Dmrt1 protein in spermatogonia and spermatocytes, which indicates plausible role in spermatogenesis. Histological analysis indicated initiation of gonadal sex differentiation in catfish around 40-50 days after hatching. The potential role for dmrt1 in testicular differentiation is evident from its stage-dependent elevated expression in developing testis. Furthermore, dimorphic expressions of dmrt1s were evident at different stages of gonadal development or recrudescence in catfish. Treatment of methyl testosterone (MT) during early stages of gonadal sex differentiation resulted in adult males. Interestingly, we also obtained MT treated fishes having ova-testis gonads. Analysis of dmrt1, sox9a, foxl2 and cyp19a1 expression patterns in MT-treated gonads revealed tissue-specific pattern. These results together suggest that multiple dmrt1s are testis-specific markers in catfish. PMID- 19293250 TI - The virtual cell--a candidate co-ordinator for 'middle-out' modelling of biological systems. AB - Understanding the functioning of biological systems depends on tackling complexity spanning spatial scales from genome to organ to whole organism. The basic unit of life, the cell, acts to co-ordinate information received across these scales and processes the myriad of signals to produce an integrated cellular response. Cells interact with and respond to other cells through direct or indirect contact, resulting in emergent structure and function of tissues and organs. Systems biology has traditionally used either a 'top-down' or 'bottom-up' approach. However, neither approach takes account of heterogeneity or 'noise', which is an inherent feature of cellular behaviour and may have significant impact on system level behaviour. We review existing approaches to modelling that use cellular automata or agent-based methodologies, where individual cells are represented as equivalent virtual entities governed by simple rules. These paradigms allow a direct one-to-one mapping between real and virtual cells that can be exploited in terms of acquiring parameters from experimental systems, or for model validation. Such models are inherently extensible and can be integrated with other modelling modalities (e.g. partial or ordinary differential equations) to model multi-scale phenomena. Alternatively, hierarchical agent models may be used to explore the functions of biological systems across temporal and spatial scales. This review examines individual-based models and the application of the paradigm to explore multi-scale phenomena in biology. In so doing, it demonstrates how cellular-based models have begun to play an important role in the development of 'middle-out' models, but with considerable potential for future development. PMID- 19293251 TI - Inhibition of Anchorage-Independent Proliferation and G0/G1 Cell-Cycle Regulation in Human Colorectal Carcinoma Cells by 4,7-Dimethoxy-5-Methyl-l,3-Benzodioxole Isolated from the Fruiting Body of Antrodia camphorate. AB - In this study, 4,7-dimethoxy-5-methyl-l,3-benzodioxole (SY-1) was isolated from three different sources of dried fruiting bodies of Antrodia camphorate (AC). AC is a medicinal mushroom that grows on the inner heartwood wall of Cinnamomum kanehirai Hay (Lauraceae), an endemic species that is used in Chinese medicine for its anti-tumor and immunomodulatory properties. In this study, we demonstrated that SY-1 profoundly decreased the proliferation of human colon cancer cells (COLO 205) through G0/G1 cell-cycle arrest (50-150 MUM) and induction of apoptosis (>150 MUM). Cell-cycle arrest induced by SY-1 was associated with a significant increase in levels of p53, p21/Cip1 and p27/Kip1, and a decrease in cyclins D1, D3 and A. In contrast, SY-1 treatment did not induce significant changes in G0/G1 phase cell-cycle regulatory proteins in normal human colonic epithelial cells (FHC). The cells were cultured in soft agar to evaluate anchorage-independent colony formation, and we found that the number of transformed colonies was significantly reduced in the SY-1-treated COLO 205 cells. These findings demonstrate for the first time that SY-1 inhibits human colon cancer cell proliferation through inhibition of cell growth and anchorage independent colony formation in soft agar. However, the detailed mechanisms of these processes remain unclear and will require further investigation. PMID- 19293252 TI - Integration of complementary and alternative medicine into family practices in Germany: results of a national survey. AB - More than two-thirds of patients in Germany use complementary and alternative medicine (CAM) provided either by physicians or non-medical practitioners ("Heilpraktiker"). There is little information about the number of family physicians (FPs) providing CAM. Given the widespread public interest in the use of CAM, this study aimed to ascertain the use of and attitude toward CAM among FPs in Germany. A postal questionnaire developed based on qualitatively derived data was sent to 3000 randomly selected FPs in Germany. A reminder letter including a postcard (containing a single question about CAM use in practice and reasons for non-particpation in the survey) was sent to all FPs who had not returned the questionnaire. Of the 3000 FPs, 1027 (34%) returned the questionnaire and 444 (15%) returned the postcard. Altogether, 886 of the 1471 responding FPs (60%) reported using CAM in their practice. A positive attitude toward CAM was indicated by 503 FPs (55%), a rather negative attitude by 127 FPs (14%). Chirotherapy, relaxation and neural therapy were rated as most beneficial CAM therapies by FPs, whereas neural therapy, phytotherapy and acupuncture were the most commonly used therapies in German family practices. This survey clearly demonstrates that CAM is highly valued by many FPs and is already making a substantial contribution to first-contact primary care in Germany. Therefore, education and research about CAM should be increased. Furthermore, with the provision of CAM by FPs, the role of non-medical CAM practitioners within the German healthcare system is to be questioned. PMID- 19293253 TI - Can routine posttransplant HPV vaccination prevent commonly occurring epithelial cancers after allogeneic stem cell transplantation? AB - The association between squamous cell carcinoma (SCC) of the oral cavity, female genital tract, and skin with human papilloma virus (HPV) subtypes is well established in the general population and in solid organ transplant recipients, but no consistent link has been reported between HPV infection and SCC after allogeneic stem cell transplantation (allo-SCT). Studies are needed to determine if SCC, the most common secondary malignancy after allo-SCT, which is linked to chronic graft versus host disease and immunosuppressive therapy, is HPV related. Consideration should be given to assessing pretransplant HPV antibodies to identify patients at risk for HPV reactivation. If a strong relationship between HPV and second malignancies after SCT exists, studies to evaluate the immunogenicity and efficacy of quadrivalent HPV vaccine (subtypes 6, 11, 16, and 18) should be considered in both male and female long-term survivors after allo SCT. PMID- 19293254 TI - Evaluation of T- and NK-cell-targeted therapies: is there a role for rituximab prophylaxis? AB - Splizimomab, an antibody that targets CD2, was studied in the treatment of T and natural killer cell lymphoma and was found to be associated with Epstein-Barr virus lymphoproliferative disease. B cell depletion may provide a platform for further evaluation of this and other promising antibody approaches that result in T cell depletion. PMID- 19293255 TI - PPM1D is a potential therapeutic target in ovarian clear cell carcinomas. AB - PURPOSE: To identify therapeutic targets in ovarian clear cell carcinomas, a chemoresistant and aggressive type of ovarian cancer. EXPERIMENTAL DESIGN: Twelve ovarian clear cell carcinoma cell lines were subjected to tiling path microarray comparative genomic hybridization and genome-wide expression profiling analysis. Regions of high-level amplification were defined and genes whose expression levels were determined by copy number and correlated with gene amplification were identified. The effects of inhibition of PPM1D were assessed using short hairpin RNA constructs and a small-molecule inhibitor (CCT007093). The prevalence of PPM1D amplification and mRNA expression was determined using chromogenic in situ hybridization and quantitative real-time reverse transcription-PCR in a cohort of pure ovarian clear cell carcinomas and on an independent series of unselected epithelial ovarian cancers. RESULTS: Array-based comparative genomic hybridization analysis revealed regions of high-level amplification on 1q32, 1q42, 2q11, 3q24-q26, 5p15, 7p21-p22, 11q13.2-q13.4, 11q22, 17q21-q22, 17q23.2, 19q12-q13, and 20q13.2. Thirty-four genes mapping to these regions displayed expression levels that correlated with copy number gains/amplification. PPM1D had significantly higher levels of mRNA expression in ovarian clear cell carcinoma cell lines harboring gains/amplifications of 17q23.2. PPM1D inhibition revealed that PPM1D expression and phosphatase activity are selectively required for the survival of ovarian clear cell carcinoma cell lines with 17q23.2 amplification. PPM1D amplification was significantly associated with ovarian clear cell carcinoma histology (P = 0.0003) and found in 10% of primary ovarian clear cell carcinomas. PPM1D expression levels were significantly correlated with PPM1D gene amplification in primary ovarian clear cell carcinomas. CONCLUSION: Our data provide strong circumstantial evidence that PPM1D is a potential therapeutic target for a subgroup of ovarian clear cell carcinomas. PMID- 19293256 TI - Small-molecule antagonists for CXCR2 and CXCR1 inhibit human melanoma growth by decreasing tumor cell proliferation, survival, and angiogenesis. AB - PURPOSE: Melanoma, the most aggressive form of skin cancer, accounts for 75% of all skin cancer-related deaths and current therapeutic strategies are not effective in advanced disease. In the current study, we have investigated the efficacy of orally active small-molecule antagonist targeting CXCR2/CXCR1. EXPERIMENTAL DESIGN: Human A375SM melanoma cells were treated with SCH-479833 or SCH-527123, and their effect on proliferation, motility, and invasion was evaluated in vitro. We examined the downstream signaling events in the cells following treatment with antagonists. For in vivo studies, A375SM cells were implanted subcutaneously into athymic nude mice followed by administration of SCH 479833, SCH-527123, or hydroxypropyl-beta-cyclodextrin (20%) orally for 21 days and their effect on tumor growth and angiogenesis was evaluated. RESULTS: Our data show that SCH-479833 or SCH-527123 inhibited the melanoma cell proliferation, chemotaxis, and invasive potential in vitro. Treatment of melanoma cells with SCH-479833 or SCH-527123 also inhibited tumor growth. Histologic and histochemical analyses showed significant (P < 0.05) decreases in tumor cell proliferation and microvessel density in tumors. Moreover, we observed a significant increase in melanoma cell apoptosis in SCH-479833- or SCH-527123 treated animals compared with controls. CONCLUSION: Together, these studies show that selectively targeting CXCR2/CXCR1 with orally active small-molecule inhibitors is a promising therapeutic approach for inhibiting melanoma growth and angiogenesis. PMID- 19293257 TI - Radioimmunotherapy of experimental human metastatic melanoma with melanin-binding antibodies and in combination with dacarbazine. AB - PURPOSE: Melanin has emerged as an attractive target for radioimmunotherapy (RIT) of melanoma, and a radiolabeled monoclonal antibody (mAb) 6D2 to melanin is currently in clinical evaluation. We investigated two approaches to improve the targeting of radiation to tumors using melanin-binding mAbs: (a) the use of an additional mAb to melanin could provide information on whether using antibodies to melanin can serve as a general approach to development of therapeutics for melanoma, and (b) as melanin targeting involves the antibody binding to extracellular melanin released from necrotic melanoma cells, we hypothesized that the administration of a chemotherapeutic agent followed by RIT would facilitate the delivery of radiation to the tumors due to the increased presence of free melanin. EXPERIMENTAL DESIGN: We evaluated the therapeutic efficacy of two melanin-binding IgM mAbs labeled with (188)Re (6D2 and 11B11). We compared the efficacy of RIT with (188)Re-6D2 to chemotherapy with dacarbazine (DTIC) and to combined chemotherapy and RIT in human metastatic melanoma-bearing nude mice. RESULTS: Therapeutic efficacy of (188)Re-labeled 6D2 and 11B11 was comparable despite differences in their affinity and binding site numbers. Comparison of chemotherapy with DTIC and RIT revealed that RIT was more effective in slowing tumor growth in mice. Administration of DTIC followed by RIT was more effective than either modality alone. CONCLUSIONS: These results provide encouragement for the development of RIT for melanoma with melanin-binding mAbs and suggest that combining chemotherapy and RIT may be a promising approach for the treatment of metastatic melanoma. PMID- 19293258 TI - Stem cells and the natural history of lung cancer: implications for lung cancer screening. AB - Lung cancer is not simply a single disease, but a collection of several phenotypically very diverse and regionally distinct neoplasias. Its natural history is complex and not yet fully understood. Stem cells and the complex interaction with the microenvironment of the tumor and the immune system play an important role in tumor progression and metastasizing capacity. This finding explains why lung cancer does not always follow the multistep carcinogenetic and exponential growth model and why small lesions do not always equate to early stage disease. Despite the fact that volume doubling times are increasingly used as surrogate markers for the natural history of lung cancer and as estimates for the proportion of overdiagnosed cases, it is only a momentary impression. At baseline screening especially, screen-detected lung cancer cases are preferably detected when they are in the indolent phase of their growth curve (length-biased sampling), from which it can by no means be concluded that they may not progress or metastasize at a later stage. Because the natural history of lung cancer is only partly elucidated, conclusions on the impact of overdiagnosis in lung cancer screening are premature. PMID- 19293259 TI - Combined PTEN and p27kip1 protein expression patterns are associated with obesity and prognosis in endometrial carcinomas. AB - PURPOSE: Phosphatase and tensin homologue deleted from chromosome 10 (PTEN) and p27(kip1) proteins are key players of the Akt pathway, which is nutritionally regulated by insulin receptor signaling and influenced by estrogens. In this study, the prognostic relevance of the PTEN/p27(kip1) protein expression in endometrial carcinoma in relationship to the body mass index (BMI) was determined. EXPERIMENTAL DESIGN: BMI and prognosis of 452 surgically treated patients with endometrial carcinoma were correlated with histologic subtype, International Federation of Gynecology and Obstetrics (FIGO) stage, and differentiation grade. The expression of PTEN and p27(kip1) was examined in 257 tumors by immunohistochemistry using a tissue microarray approach. RESULTS: Lack of PTEN was observed in 136 of 257 (53%) tumors and absence of p27(kip1) expression was observed in 106 of 225 (47%) tumors. Absence of both proteins was significantly associated with well-differentiated tumors [PTEN (P < 0.02) and p27(kip1) (P < 0.009)]. Differentiation grade, tumor stage, and histologic type were independent of an increased BMI. Importantly, tumors of obese women expressed significantly less PTEN (P < 0.008) and less p27(kip1) (P < 0.01) than tumors from nonobese patients. Combined absence of both PTEN and p27(kip1) expression characterized a group of 75 (32%) tumors with favorable clinical outcome, particularly in the FIGO stages I and II (P = 0.003) of obese patients. Cox regression analysis revealed that PTEN/p27(kip1) phenotype, FIGO stage, and histologic grade were independent predictors of prognosis in endometrioid endometrial carcinoma. CONCLUSIONS: Inactivation of PTEN/p27(kip1) proteins is a specific feature in the progression of endometrial carcinoma in obese patients. The phenotype of the combined loss of PTEN/p27(kip1) protein expression in obese patients is associated with a significantly better prognosis in endometrioid endometrial carcinoma. PMID- 19293260 TI - EBV-related lymphoproliferative disease complicating therapy with the anti-CD2 monoclonal antibody, siplizumab, in patients with T-cell malignancies. AB - PURPOSE: We report an increased incidence of EBV-induced B-cell lymphoproliferative disease (LPD) in patients treated with siplizumab, an anti CD2 antibody. The development of EBV-LPD has been associated with the use of immunosuppressive agents used in solid organ, bone marrow, and stem cell transplantation and in certain congenital immunodeficiencies. EXPERIMENTAL DESIGN: We conducted a single-institution phase I dose-escalation trial of siplizumab, a humanized monoclonal antibody to CD2, in 29 patients with T-cell malignancies. RESULTS: Although initial responses were encouraging, 4 (13.7%) patients developed EBV-LPD and the trial was stopped. Reductions in CD4(+) and CD8(+) cell count numbers in response to therapy were seen in all patients, but in those patients developing EBV-LPD a significantly greater reduction in natural killer (NK) cell number and CD2 expression on T cells was seen. These findings highlight the importance of NK-cell depletion and CD2 expression in addition to T cell depletion in the etiology of EBV-LPD. CONCLUSIONS: The emergence of EBV-LPD may be associated with the ability of siplizumab to deplete both T and NK cells without affecting B cells. Agents that deplete T- and NK-cell populations without affecting B cell number should be screened for this potentially serious adverse event. PMID- 19293261 TI - Testosterone and growth hormone improve body composition and muscle performance in older men. AB - CONTEXT: Impairments in the pituitary-gonadal axis with aging are associated with loss of muscle mass and function and accumulation of upper body fat. OBJECTIVES: We tested the hypothesis that physiological supplementation with testosterone and GH together improves body composition and muscle performance in older men. DESIGN, SETTING, AND PARTICIPANTS: One hundred twenty-two community-dwelling men 70.8 +/- 4.2 yr of age with body mass index of 27.4 +/- 3.4 kg/m2, testosterone of 550 ng/dl or less, and IGF-I in lower adult tertile (< or =167 ng/dl) were randomized to receive transdermal testosterone (5 or 10 g/d) during a Leydig cell clamp plus GH (0, 3, or 5 microg/kg . d) for 16 wk. MAIN OUTCOME MEASURES: Body composition by dual-energy x-ray absorptiometry, muscle performance, and safety tests were conducted. RESULTS: Total lean body mass increased (1.0 +/- 1.7 to 3.0 +/- 2.2 kg) as did appendicular lean tissue (0.4 +/- 1.4 to 1.5 +/- 1.3 kg), whereas total fat mass decreased by 0.4 +/- 0.9 to 2.3 +/- 1.7 kg as did trunk fat (0.5 +/- 0.9 to 1.5 +/- 1.0 kg) across the six treatment groups and by dose levels for each parameter (P < or = 0.0004 for linear trend). Composite maximum voluntary strength of upper and lower body muscles increased by 14 +/- 34 to 35 +/- 31% (P < 0.003 in the three highest dose groups) that correlated with changes in appendicular lean mass. Aerobic endurance increased in all six groups (average 96 +/- 137 sec longer). Systolic and diastolic blood pressure increased similarly in each group with mean increases of 12 +/- 14 and 8 +/- 8 mm Hg, respectively. Other predictable adverse events were modest and reversible. CONCLUSIONS: Supplemental testosterone produced significant gains in total and appendicular lean mass, muscle strength, and aerobic endurance with significant reductions in whole-body and trunk fat. Outcomes appeared to be further enhanced with GH supplementation. PMID- 19293262 TI - Multicenter contraceptive efficacy trial of injectable testosterone undecanoate in Chinese men. AB - CONTEXT: Hormonal male contraceptive regimens effectively and reversibly suppress sperm production, but there are few large-scale efficacy studies. OBJECTIVE: The safety, contraceptive efficacy, reversibility, and feasibility of injectable testosterone undecanoate (TU) in tea seed oil as a hormonal male contraceptive was assessed. DESIGN: This was a multicenter, phase III, contraceptive efficacy clinical trial. PARTICIPANTS: A total of 1045 healthy fertile Chinese men were recruited throughout China into the study. INTERVENTION(S): Injections of 500 mg TU were administered monthly for 30 months. A definition of severe oligozoospermia (< or =1 x 10(6)/ml) was used as a criterion of spermatogenic suppression and as the threshold for entering the contraceptive efficacy phase. MAIN OUTCOME MEASURE(S): The primary outcome was pregnancy rate in the partner. Other outcomes include: semen parameters, testis volumes, reproductive hormone levels, and safety laboratory tests. RESULTS: Forty-three participants (4.8%) did not achieve azoospermia or severe oligozoospermia within the 6-month suppression phase. A total of 855 participants entered into the efficacy phase, and 733 participants completed monthly TU treatment and follow-up. There were nine pregnancies in 1554.1 person-years of exposure in the 24-month efficacy phase for a cumulative contraceptive failure rate of 1.1 per 100 men. The combined method failure rate was 6.1%, comprising 4.8% with inadequate suppression and 1.3% with postsuppression sperm rebound. No serious adverse events were reported. Spermatogenesis returned to the normal fertile reference range in all but two participants. CONCLUSIONS: Monthly injection of 500 mg TU provides safe, effective, reversible, and reliable contraception in a high proportion of healthy fertile Chinese men. PMID- 19293263 TI - Normoalbuminuric type 1 diabetic patients with retinopathy have an impaired tubular response to desmopressin: its relationship with plasma endothelin-1. AB - OBJECTIVE: The aim of the study was to evaluate whether normoalbuminuric type 1 diabetic patients with diabetic retinopathy (DR) have an impaired tubular response to desmopressin (dDAVP, a synthetic analog of vasopressin) administration, and its relationship with plasma and urine endothelin-1 (ET-1) levels. DESIGN: This was an interventional case-control study. SETTING: The study was conducted at a referral center. PARTICIPANTS: Fifteen normoalbuminuric type 1 diabetic patients with DR were compared with 30 normoalbuminuric type 1 diabetic patients without DR. Both groups were matched by age, gender, body mass index, glycosylated hemoglobin, and the main laboratory markers of kidney function. INTERVENTION: After a 12-h period of water deprivation, dDAVP (0.3 microg/kg) was infused over 20 min. Urine was collected at baseline and 1, 2, and 3 h after dDAVP administration. ET-1 was assessed by ELISA. RESULTS: dDAVP induced a lower rise in urine osmolality in patients with DR (from 650 +/- 206 to 754 +/- 224 mosmol/kg; P = 0.02) than in diabetic patients without DR (from 714 +/- 194 to 905 +/- 163 mosmol/kg; P < 0.0001). In addition, fractional excretion of Na+ decreased in patients without DR (from 0.45 +/- 0.30 to 0.29 +/- 0.29%; P = 0.04) but not in the diabetic patients with DR (from 0.36 +/- 0.22 to 0.36 +/- 0.40%; P = 0.96). Plasma ET-1 levels were inversely correlated with the response of urinary osmolality after dDAVP administration (r = -0.62; P = 0.008). CONCLUSIONS: Normoalbuminuric type 1 diabetic patients with DR have impaired renal response to dDAVP that is related to plasma ET-1 levels. Further studies are required to elucidate whether this tubular resistance to dDAVP might favor dehydration in these patients. PMID- 19293264 TI - Blood pressure in pediatric patients with Cushing syndrome. AB - CONTEXT: Hypertension (HTN) has been reported in up to 60% of children with Cushing syndrome (CS), but its course, side effects, and potential differences among various causes of CS have not been adequately studied. OBJECTIVE: The objective of the study was to measure blood pressure in pediatric patients with CS before and after transphenoidal surgery or adrenalectomy and identify side effects and rates of residual HTN. DESIGN: Data from 86 children with corticotropinomas [Cushing disease (CD)] and 27 children with ACTH-independent CS (AICS) were analyzed. RESULTS: Patients with CD and AICS had significant HTN before surgery; more patients with AICS had systolic HTN (SHTN) than with CD (74 vs. 44%, P = 0.0077), but the rate of diastolic HTN (DHTN) was similar. Both groups experienced significant decreases in SHTN immediately after transphenoidal surgery and adrenalectomy. One year postoperatively, both SHTN and DHTN were lower than the preoperative values in all patients, but as many as 16 and 4% of the patients with CD and 21 and 5% of the patients with AICS still had SHTN and DHTN, respectively. Higher blood pressure preoperatively correlated with cortisol levels. Two patients suffered serious side effects: one with multiple infarcts and another with hypertensive encephalopathy. CONCLUSIONS: Children with CS are at risk for residual HTN despite a significant improvement after surgical cure. HTN appears to correlate with the degree of hypercortisolemia. Serious HTN related side effects, although rare, may occur during the perioperative period. PMID- 19293265 TI - Type 2 iodothyronine deiodinase in skeletal muscle: effects of hypothyroidism and fasting. AB - CONTEXT: The iodothyronine deiodinases D1, D2, and D3 enable tissue-specific adaptation of thyroid hormone levels in response to various conditions, such as hypothyroidism or fasting. The possible expression of D2 mRNA in skeletal muscle is intriguing because this enzyme could play a role in systemic as well as local T3 production. OBJECTIVE: We determined D2 activity and D2 mRNA expression in human skeletal muscle biopsies under control conditions and during hypothyroidism, fasting, and hyperinsulinemia. DESIGN: This was a prospective study. SETTING: The study was conducted at a university hospital. PATIENTS: We studied 11 thyroidectomized patients with differentiated thyroid carcinoma (DTC) on and after 4 wk off T4( replacement and six healthy lean subjects in the fasting state and during hyperinsulinemia after both 14 and 62 h of fasting. MEAN OUTCOME MEASURES: D2 activity and D2 mRNA levels were measured in skeletal muscle samples. RESULTS: No differences were observed in muscle D2 mRNA levels in DTC patients on and off T4 replacement therapy. In healthy subjects, muscle D2 mRNA levels were lower after 62 h compared to 14 h of fasting. Insulin increased mRNA expression after 62 h, but not after 14 h of fasting. Skeletal muscle D2 activities were very low and not influenced by hypothyroidism and fasting. CONCLUSION: Human skeletal muscle D2 mRNA expression is modulated by fasting and insulin, but not by hypothyroidism. The lack of a clear effect of D2 mRNA modulation on the observed low D2 activities questions the physiological relevance of D2 activity in human skeletal muscle. PMID- 19293266 TI - Inhibition of Src with AZD0530 reveals the Src-Focal Adhesion kinase complex as a novel therapeutic target in papillary and anaplastic thyroid cancer. AB - CONTEXT: Focal adhesion kinase (FAK) and Src are overexpressed and activated in many cancers and have been associated with tumor progression. The role of the Src FAK complex has not been characterized in papillary and anaplastic thyroid cancer (PTC and ATC). OBJECTIVE: The goal of this study was to determine the role of Src and FAK in the growth and invasion of PTC and ATC. DESIGN: PTC and ATC cells were treated with the oral Src inhibitor, AZD0530, to determine the consequences of Src inhibition using growth and invasion assays. FAK and phospho-FAK levels were analyzed in cell lines as well as in PTC tumor samples. RESULTS: AZD0530 treatment inhibited the growth and invasion in four of five thyroid cancer cell lines, and inhibition did not correlate with basal levels of phospho-Src. Instead, we show for the first time that FAK, a critical substrate and effector of Src, is phosphorylated at tyrosine residue 861 (pY861) in PTC and ATC cells, and high levels of phospho-FAK correlate with AZD0530 sensitivity. We further showed that pY861-FAK phosphorylation is Src-dependent. Sensitivity to AZD0530 was confirmed using a preclinical three-dimensional culture model. Phospho-ERK1/2 was not affected by AZD0530, indicating that Src signaling does not require MAPK. Finally, FAK and pY861-FAK were expressed in 10 of 10 and five of 10 PTC tumors, respectively. CONCLUSIONS: Inhibition of the Src-FAK complex represents a promising therapeutic strategy for patients with advanced thyroid cancer, and phospho-FAK represents a potential biomarker for response. PMID- 19293267 TI - Autoantibodies in type 2 diabetes induce stress fiber formation and apoptosis in endothelial cells. AB - CONTEXT: Macular edema contributes to visual impairment, and albuminuria is associated with increased cardiovascular mortality in adults with type 2 diabetes mellitus. These microvascular complications result from increased capillary leakage of plasma proteins whose causation is not completely understood. OBJECTIVE: The objective of the present study was to test whether plasma from type 2 diabetes with maculopathy/albuminuria or control subjects contains autoantibodies that can induce apoptosis or activate Rho kinase (ROCK) in endothelial cells. DESIGN: A cohort of Veterans Affairs Diabetes Trial adults (>40 yr of age) was randomized to standard vs. intensive glycemic treatment lasting 5-7.5 yr. SETTING: The study was conducted in outpatient clinics. PATIENTS: Case and age-matched control subjects who differed for the baseline presence of significant diabetic maculopathy and/or progression to macro albuminuria were included in the study. INTERVENTION: Pharmacological and lifestyle interventions in the Veterans Affairs Diabetes Trial generally resulted in substantially improved glycemic, blood pressure, and lipid levels. RESULTS: Autoantibodies from patients with macular edema or progression to albuminuria potently induced caspase-dependent apoptosis in endothelial cells (up to 60%), whereas IgG from age-matched normal plasma caused much less apoptosis (<10%; P < 0.0001). The active inhibitory autoantibodies triggered stress fiber formation in endothelial cells likely through the activation of Rho guanosine 5' triphosphatase, which could be nearly completed inhibited by 10 microm Y27632, a specific ROCK inhibitor. CONCLUSIONS: These results suggest that autoantibodies from a subset of advanced type 2 diabetes may contribute to diabetic vascular complications by activating ROCK, inducing stress fiber formation and apoptosis in endothelial cells. PMID- 19293268 TI - Mutations in regulatory subunit type 1A of cyclic adenosine 5'-monophosphate dependent protein kinase (PRKAR1A): phenotype analysis in 353 patients and 80 different genotypes. AB - BACKGROUND: The "complex of myxomas, spotty skin pigmentation, and endocrine overactivity," or "Carney complex" (CNC), is caused by inactivating mutations of the regulatory subunit type 1A of the cAMP-dependent protein kinase (PRKAR1A) gene and as yet unknown defect(s) in other gene(s). Delineation of a genotype phenotype correlation for CNC patients is essential for understanding PRKAR1A function and providing counseling and preventive care. METHODS: A transatlantic consortium studied the molecular genotype and clinical phenotype of 353 patients (221 females and 132 males, age 34 +/- 19 yr) who carried a germline PRKAR1A mutation or were diagnosed with CNC and/or primary pigmented nodular adrenocortical disease. RESULTS: A total of 258 patients (73%) carried 80 different PRKAR1A mutations; 114 (62%) of the index cases had a PRKAR1A mutation. Most PRKAR1A mutations (82%) led to lack of detectable mutant protein (nonexpressed mutations) because of nonsense mRNA mediated decay. Patients with a PRKAR1A mutation were more likely to have pigmented skin lesions, myxomas, and thyroid and gonadal tumors; they also presented earlier with these tumors. Primary pigmented nodular adrenocortical disease occurred earlier, was more frequent in females, and was the only manifestation of CNC with a gender predilection. Mutations located in exons were more often associated with acromegaly, myxomas, lentigines, and schwannomas, whereas the frequent c.491 492delTG mutation was commonly associated with lentigines, cardiac myxomas, and thyroid tumors. Overall, nonexpressed PRKAR1A mutations were associated with less severe disease. CONCLUSION: CNC is genetically and clinically heterogeneous. Certain tumors are more frequent, with specific mutations providing some genotype phenotype correlation for PRKAR1A mutations. PMID- 19293269 TI - Fetal and postnatal growth and body composition at 6 months of age. AB - OBJECTIVES: The objectives of the study was to examine which parental, fetal, and postnatal characteristics are associated with fat and lean mass at the age of 6 months and examine the effect of growth (catch-down, catch-up) in fetal life and early infancy on fat and lean mass. DESIGN: This study was embedded in the Generation R Study, a prospective cohort study from early fetal life onward. Body composition was measured by dual-energy X-ray absorptiometry in 252 infants at 6 months. Parental, fetal, and postnatal data were collected by physical and fetal ultrasound examinations and questionnaires. RESULTS: Children with fetal catch-up in weight (gain in weight sd score >0.67) in the second trimester tended to have a higher fat mass percentage [FM(%)] at 6 months of age, whereas children with fetal catch-down in weight had a lower FM(%) compared with nonchangers. In the third trimester, both catch-up and catch-down in weight were associated with an increase in FM(%) at 6 months. Children with catch-down in the third trimester had a greater risk for postnatal catch-up in weight greater than 0.67 sd score. Birth weight and weight at 6 wk were positively associated with fat mass at 6 months. Postnatal catch-up in weight within 6 wk after birth had the highest association with total and truncal FM(%) at 6 months. Total and truncal FM were higher in girls. CONCLUSION: Catch-down in weight in the third trimester was strongly associated with postnatal catch-up within 6 wk after birth, and both were associated with an increase in fat mass at the age of 6 months. Our study shows that fetal as well as postnatal growth patterns are associated with body composition in early childhood. PMID- 19293270 TI - Expression analysis of dopamine receptor subtypes in normal human pituitaries, nonfunctioning pituitary adenomas and somatotropinomas, and the association between dopamine and somatostatin receptors with clinical response to octreotide LAR in acromegaly. AB - CONTEXT: Dopamine receptor (DR) and somatostatin receptor subtype expression in pituitary adenomas may predict the response to postsurgical therapies. OBJECTIVES: Our objectives were to assess and compare the mRNA levels of DR1-5 and somatostatin receptors 1-5 in normal pituitaries (NPs), nonfunctioning pituitary adenomas (NFPAs), and somatotropinomas. In addition, we determined whether the level of DR expression correlates with the in vivo response to octreotide-LAR in acromegalic patients. DESIGN AND PATIENTS: Eight NPs, 30 NFPAs, and 39 somatotropinomas were analyzed for receptor mRNA levels by real-time RT PCR. The DR2 short variant was estimated as the DR2 long/DR2 total (DR2T). The relationship between DR expression and the postsurgical response to octreotide LAR was assessed in 19 of the acromegalic patients. RESULTS: DR3 was not detected. The relationship between expression levels of DR subtypes in NPs and somatotropinomas was DR2T>>>DR4>>DR5>DR1, whereas in NFPAs, DR2T>>>DR4>>DR1>DR5. The DR2 short variant was the predominant DR2 variant in the majority of samples. In acromegalics treated with octreotide-LAR, DR1 was negatively correlated with percent GH reduction (3 months: r = -0.67, P = 0.002; and 6 months: r = -0.58, P = 0.009), and DR5 was positively correlated with percent IGF-I reduction (3 months: r = 0.55, P = 0.01; and 6 months: r = 0.47, P = 0.04). CONCLUSIONS: DR2 is the predominant DR subtype in NPs, NFPAs, and somatotropinomas. The fact that DR1, DR4, and DR5 are also expressed in many adenomas tested suggests that these receptors might also play a role in the therapeutic impact of postsurgical medical therapies in patients with NFPA and acromegaly. This was supported by the finding that the in vivo response to octreotide-LAR was negatively associated with DR1 and positively associated with DR5. PMID- 19293271 TI - Bone mineral density, growth, and thyroid function in long-term survivors of pediatric Hodgkin's lymphoma treated with chemotherapy only. AB - BACKGROUND: The aim of this study was to investigate the long-term side effects of treatment for childhood Hodgkin's lymphoma with chemotherapy only on growth, bone mineral density (BMD), body composition, and thyroid function. PROCEDURE: A total of 88 patients (56 male, 32 female; 17.6-42.6 yr), treated for childhood Hodgkin's lymphoma from 1974-1998 with combination chemotherapy adriamycin (doxorubicin), bleomycin, vinblastine, and dacarbazine or epirubicin, bleomycin, vinblastine, dacarbazine with or without mechlorethamine, oncovin (vincristine), procarbazine, and prednisone (MOPP) with the intention to avoid radiotherapy, participated in this study. Median follow-up was 15.5 yr (range 5.6-30.2). BMD of lumbar spine and total body (BMD-TB), and body composition were measured using dual-energy x-ray absorptiometry. Bone mineral apparent density of the lumbar spine was calculated to correct for bone size. Free T4 and TSH were measured. RESULTS: Men treated with MOPP had a significantly reduced height with normal body proportions. Women treated with MOPP had decreased BMD-TB and bone mineral apparent density of the lumbar spine as compared with healthy controls. Percent body fat was significantly increased in female patients treated without MOPP. Body mass index was significantly increased in male patients treated without MOPP, whereas lean body mass was normal in all patients. All patients, except one, treated with chemotherapy only had normal thyroid function. However, five patients who received additional radiation to the thyroid either had abnormal levels of TSH or free T4, or used thyroid hormones. CONCLUSIONS: Lean body mass was normal in all patients; thyroid function was normal in all but one patient. The use of MOPP leads to decreased height and increased body mass index in men and decreased BMD-TB in women. PMID- 19293272 TI - Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization. AB - The effects of photocaged nucleosides on the DNA polymerization reaction was investigated, finding that most polymerases are unable to recognize and read through the presence of a single caging group on the DNA template. Based on this discovery, a new method of introducing mutations into plasmid DNA via a light mediated mutagenesis protocol was developed. This methodology is advantageous over several common approaches in that it requires the use of only two polymerase chain reaction primers, and does not require any restriction sites or use of restriction enzymes. Additionally, this approach enables not only site-directed mutations, but also the insertion of DNA strands of any length into plasmids and the deletion of entire genes from plasmids. PMID- 19293273 TI - Expression profiling of hypothetical genes in Desulfovibrio vulgaris leads to improved functional annotation. AB - Hypothetical (HyP) and conserved HyP genes account for >30% of sequenced bacterial genomes. For the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough, 347 of the 3634 genes were annotated as conserved HyP (9.5%) along with 887 HyP genes (24.4%). Given the large fraction of the genome, it is plausible that some of these genes serve critical cellular roles. The study goals were to determine which genes were expressed and provide a more functionally based annotation. To accomplish this, expression profiles of 1234 HyP and conserved genes were used from transcriptomic datasets of 11 environmental stresses, complemented with shotgun LC-MS/MS and AMT tag proteomic data. Genes were divided into putatively polycistronic operons and those predicted to be monocistronic, then classified by basal expression levels and grouped according to changes in expression for one or multiple stresses. One thousand two hundred and twelve of these genes were transcribed with 786 producing detectable proteins. There was no evidence for expression of 17 predicted genes. Except for the latter, monocistronic gene annotation was expanded using the above criteria along with matching Clusters of Orthologous Groups. Polycistronic genes were annotated in the same manner with inferences from their proximity to more confidently annotated genes. Two targeted deletion mutants were used as test cases to determine the relevance of the inferred functional annotations. PMID- 19293274 TI - Light-up properties of complexes between thiazole orange-small molecule conjugates and aptamers. AB - The full understanding of dynamics of cellular processes hinges on the development of efficient and non-invasive labels for intracellular RNA species. Light-up aptamers binding fluorogenic ligands show promise as specific labels for RNA species containing those aptamers. Herein, we took advantage of existing, non light-up aptamers against small molecules and demonstrated a new class of light up probes in vitro. We synthesized two conjugates of thiazole orange dye to small molecules (GMP and AMP) and characterized in vitro their interactions with corresponding RNA aptamers. The conjugates preserved specific binding to aptamers while showing several 100-fold increase in fluorescence of the dye (the 'light up' property). In the presence of free small molecules, conjugates can be displaced from aptamers serving also as fluorescent sensors. Our in vitro results provide the proof-of-concept that the small-molecule conjugates with light-up properties can serve as a general approach to label RNA sequences containing aptamers. PMID- 19293275 TI - AEBP2 as a potential targeting protein for Polycomb Repression Complex PRC2. AB - AEBP2 is a zinc finger protein that has been shown to interact with the mammalian Polycomb Repression Complex 2 (PRC2). In the current study, we characterized this unknown protein and tested its potential targeting roles for the PRC2. AEBP2 is an evolutionarily well-conserved gene that is found in the animals ranging from flying insects to mammals. The transcription of mammalian AEBP2 is driven by two alternative promoters and produces at least two isoforms of the protein. These isoforms show developmental stage-specific expression patterns: the adult specific larger form (51 kDa) and the embryo-specific smaller form (32 kDa). The AEBP2 protein binds to a DNA-binding motif with an unusual bipartite structure, CTT(N)15-23cagGCC with lower-case being less critical. A large fraction of AEBP2's target loci also map closely to the known target loci of the PRC2. In fact, many of these loci are co-occupied by the two proteins, AEBP2 and SUZ12. This suggests that AEBP2 is most likely a targeting protein for the mammalian PRC2 complex. PMID- 19293276 TI - The solution structure of the first PHD finger of autoimmune regulator in complex with non-modified histone H3 tail reveals the antagonistic role of H3R2 methylation. AB - Plant homeodomain (PHD) fingers are often present in chromatin-binding proteins and have been shown to bind histone H3 N-terminal tails. Mutations in the autoimmune regulator (AIRE) protein, which harbours two PHD fingers, cause a rare monogenic disease, autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). AIRE activates the expression of tissue-specific antigens by directly binding through its first PHD finger (AIRE-PHD1) to histone H3 tails non methylated at K4 (H3K4me0). Here, we present the solution structure of AIRE-PHD1 in complex with H3K4me0 peptide and show that AIRE-PHD1 is a highly specialized non-modified histone H3 tail reader, as post-translational modifications of the first 10 histone H3 residues reduce binding affinity. In particular, H3R2 dimethylation abrogates AIRE-PHD1 binding in vitro and reduces the in vivo activation of AIRE target genes in HEK293 cells. The observed antagonism by R2 methylation on AIRE-PHD1 binding is unique among the H3K4me0 histone readers and represents the first case of epigenetic negative cross-talk between non methylated H3K4 and methylated H3R2. Collectively, our results point to a very specific histone code responsible for non-modified H3 tail recognition by AIRE PHD1 and describe at atomic level one crucial step in the molecular mechanism responsible for antigen expression in the thymus. PMID- 19293277 TI - From Amrita to substance D: psychopharmacology, political economy, and technologies of the self. PMID- 19293278 TI - Trussed in evidence? Ambiguities at the interface between clinical evidence and clinical practice. AB - This article considers the dominance that randomized controlled trials (RCTs) of psychotropic agents currently have in relation to the practice of psychiatry in mental health and primary care settings. In contemporary psychiatry, data of marginal significance based on rating scale measures are privileged as evidence that treatments are effective, while judgments of drug effects based on clinical practice are downgraded. The dominance of RCTs has also led to an increasing promotion of rating scales in clinical practice, described here as ;rating scale mongering.' The logical consequence of current interpretations of RCT data is that clinicians should adhere to guidelines which are based on a systematic assembly of such data, but the selective publication of trial data and ghostwriting of publications, lays the basis for guideline capture, and a corresponding capture of evidence-based clinical practice by pharmaceutical companies. PMID- 19293279 TI - From descartes to desipramine: psychopharmacology and the self. AB - Despite the remarkably widespread use of the new generation of antidepressants, almost everything we know about their effects comes from animal studies and clinical trials in which the sole parameter of interest is depressive symptomatology. Almost nothing is known about the effects that antidepressants have on cognition, affect, or motivation when used over a period of months or years. Nor do we understand what effects, if any, antidepressants have on what we think of as the self. In this article, we argue that neither psychiatry nor philosophy, in their current state, are well equipped to think about these issues. In order to explore this idea, we consider the neurobiology of romantic love and its relation to antidepressant neurochemistry. This case study, we suggest, supports the view that antidepressants are very likely to have significant effects on personhood as well as the suggestion that we are in need of new ways of thinking about the self and its pathologies. PMID- 19293280 TI - Pills that swallow policy: clinical ethnography of a Community Mental Health Program in northern India. AB - India's National Mental Health Program (NMHP) was initiated in 1982 with the objective of promoting community participation and accessible mental health services. A key component involves central government calculation and funding for psychotropic medication. Based on clinical ethnography of a community psychiatry program in north India, this article traces the biosocial journey of psychotropic pills from the centre to the periphery. As the pill journeys from the Ministry of Health to the clinic, its symbolic meaning transforms from an emphasis on accessibility and participation to the administration of a discrete ;treatment.' Instead of embodying participation and access, the pill achieves the opposite: silencing community voices, re-enforcing existing barriers to care, and relying on pharmacological solutions for psychosocial problems. The symbolic inscription of NMHP policies on the pill fail because they are undercut by more powerful meanings generated from local cultural contexts. An understanding of this process is critical for the development of training and policy that can more effectively address local mental health concerns in rural India. PMID- 19293281 TI - The unlicensed lives of antidepressants in India: generic drugs, unqualified practitioners, and floating prescriptions. AB - Antidepressant uses have been rising rapidly over the past decades. Two main theories have been advanced to explain this. One claims that socio-economic change causes a global rise of depressive illness. The other holds that European and North American corporations are aggressively marketing antidepressants to expand their global reach. Both theories assume that multinational capitalism drives rising depression rates. Based on ethnographic data from India, this article shows that antidepressants are increasingly used in this country as well, but for reasons than have been little explored yet. Taking fluoxetine (Prozac) as the main example, it is argued that the spread of antidepressants in India is ;unlicensed' by Euro-American corporations in at least three ways: (i) drug marketing is driven by Indian generic producers; (ii) fluoxetine is given by practitioners who have no license to do so; and (iii) knowledge of fluoxetine is spread through unlicensed ;floating' prescriptions that patients take from one prescriber to another. PMID- 19293282 TI - 'Consumers are patients!' shared decision-making and treatment non-compliance as business opportunity. AB - This article describes an aspect of the progressive insertion of commercial interests into the relationship between patients and their clinicians, with particular reference to psychiatry. Treatment noncompliance, a long-standing problem for healthcare professionals, has lately drawn the attention of the pharmaceutical and allied industries as a site at which to improve return on investment (ROI). Newly founded corporate ;compliance departments' and specialized consultancies that regard noncompliance as a form of marketing failure are seeking to rectify it with reinvigorated models and strategies. This intervention stands to impact patients' experience of illness as well as the participation of those formally (physicians, case managers, etc.) and informally (family, friends, etc.) involved in treatment. My analysis draws upon observation at compliance conferences to demonstrate the contrasting models of patient empowerment underlying the marketing vs. medical approaches. I propose a research agenda for measuring the effects of industry compliance programs. PMID- 19293283 TI - Uncanny scripts: understanding pharmaceutical emplotment in the aboriginal context. AB - This article outlines a new social reality of global psycho-pharmaceutical prescribing: the pharmaceutical family, or ;phamily.' Ethnographic case studies from Manitoba, Canada (2002 to 2004) show how pharmaceutical emplotment, involving a synergy between cultural and drug scripts, can have uncanny consequences for vulnerable groups, such as Aboriginal children. Observations and interview transcripts of high prescribing doctors are analyzed to understand the prescribing logic of using psychoactive medication, such as methylphenidate, in young Aboriginal children diagnosed with FASD and/or ADHD. Pharmaceutical narratives are presented in order to show how non-compliance to psychotropic prescribing can further marginalize Aboriginal children and is related to the history of colonial practices in Canada. PMID- 19293284 TI - Adolescent experience of psychotropic treatment. AB - Despite growing concern over the treatment of adolescents with psychiatric medications, little research has examined youth understandings and interpretations of mental illness and psychotropic treatment. This article reports the exploratory findings of semi-structured and open-ended interviews carried out with 20 adolescents diagnosed with one or more psychiatric disorders, and who were currently prescribed psychiatric medications. Grounded theory coding procedures were used to identify themes related to adolescent subjective experience with psychiatric medications. The categories identified are interpreted as different points of view through which adolescents understand and take action upon their illness concerns; their need for medication treatment; their perceptions of how medications work; their responses to parental and other influences upon medication treatment; and, their everyday management activities. PMID- 19293285 TI - Tense prescriptions? Alzheimer medications and the anthropology of uncertainty. AB - This article discusses the two major groups of Alzheimer medications, which are hotly debated in the specialized literature because of their doubtful efficacy. Examining this issue under the rubric of an ;anthropology of uncertainty,' this article seeks to address the question: how do doctors prescribe medications given tensions created by uncertainty? A partial answer is drawn from research conducted in Brazil with local psychogeriatricians, which has documented a high degree of certainty regarding Alzheimer drugs and their benefits. I argue that one reason for this certainty is that ;efficacy' has become increasingly non specific in Alzheimer's disease through the broadening of outcome measures in clinical trials. While such measures previously focused on cognitive symptoms, they now encompass concepts such as functionality, quality of life and activities of daily living. The certainty of the Brazilian psychogeriatricians is further buttressed by three interacting elements: (i) the influence of the pharmaceutical industry; (ii) long-standing arguments for including non-cognitive symptoms in dementia care and research; and (iii) a specific discourse found in geriatrics and gerontology, which recognizes ;the person beyond cognition.' PMID- 19293286 TI - Classifying sexual offenders: an empirical model for generating type-specific approaches to intervention. AB - Although recidivism is lower among sexual offenders who have received treatment than those who have not, evaluation and treatment studies reach different numbers on recidivism rates and effectiveness. The question still remains as to what exactly works for whom; that is, does one kind of treatment work for all sexual offenders? This empirical study attempts to develop a sex offender typology that refers to treatment-relevant factors and characteristics to generate a framework for developing specific treatment approaches. In a first step the authors conduct a survey of experts to identify treatment-relevant factors for the later typology. In the main part of the study, 199 sexual offenders incarcerated in German prison and forensic facilities are examined according to the identified characteristics. A cluster analysis results in five subtypes of sexual offenders. In this article, the clusters are presented with regard to their criminologically relevant characteristics, and implications for type-related interventions are discussed. PMID- 19293287 TI - MicroRNA-125b is a novel negative regulator of p53. AB - The p53 transcription factor is a key tumor suppressor and a central regulator of the stress response. To ensure a robust and precise response to cellular signals, p53 gene expression must be tightly regulated from the transcriptional to the post-translational levels. Computational predictions suggest that several microRNAs are involved in the post-transcriptional regulation of p53. Here we demonstrate that miR-125b, a brain-enriched microRNA, is a bona fide negative regulator of p53 in both zebrafish and humans. miR-125b-mediated down-regulation of p53 is strictly dependent on the binding of miR-125b to a microRNA response element in the 3' untranslated region of p53 mRNA. Overexpression of miR-125b represses the endogenous level of p53 protein and suppresses apoptosis in human neuroblastoma cells and human lung fibroblast cells. In contrast, knockdown of miR-125b elevates the level of p53 protein and induces apoptosis in human lung fibroblasts and in the zebrafish brain. This phenotype can be rescued significantly by either an ablation of endogenous p53 function or ectopic expression of miR-125b in zebrafish. Interestingly, miR-125b is down-regulated when zebrafish embryos are treated with gamma-irradiation or camptothecin, corresponding to the rapid increase in p53 protein in response to DNA damage. Ectopic expression of miR-125b suppresses the increase of p53 and stress-induced apoptosis. Together, our study demonstrates that miR-125b is an important negative regulator of p53 and p53-induced apoptosis during development and during the stress response. PMID- 19293289 TI - Goal setting in rehabilitation: an overview of what, why and how. PMID- 19293288 TI - Evidence that androgen-independent stromal growth factor signals promote androgen insensitive prostate cancer cell growth in vivo. AB - Activation of tumor-stromal interactions is considered to play a critical role in the promotion of tumorigenesis. To discover new therapeutic targets for hormone refractory prostate tumor growth under androgen ablation therapy, androgen sensitive LNCaP cells and the derived sublines, E9 (androgen-low-sensitive), and AIDL (androgen-insensitive), were recombined with androgen-dependent embryonic rat urogenital sinus mesenchyme (UGM). Tumors of E9 + UGM and AIDL + UGM were approximately three times as large as those of LNCaP + UGM. Tumors grown in castrated hosts exhibited reduced growth as compared with those in intact hosts. However, in castrated hosts, E9 + UGM and AIDL + UGM tumors were still approximately twice as large as those of LNCaP + UGM. Cell proliferation in tumors of E9 + UGM and AIDL + UGM grown in castrated host, was significantly higher than that in tumors of LNCaP + UGM. In vitro, expression of fibroblast growth factor (FGF)-2 and IGF-I, but not FGF-7 mRNA, was significantly reduced in UGM under androgen starvation. In cell culture, E9 cells were responsive to FGF-2 and FGF-7 stimulation, while AIDL responded to FGF-7 and IGF-1. Expression of FGFR1 and FGFR2 was considerably higher in E9 than those in LNCaP, similarly expression of FGFR2 and IGF-IR were elevated in AIDL. These data suggest that activation of prostate cancer cell growth through growth factor receptor expression may result in the activity of otherwise androgen-independent stromal growth factor signals such as FGF-7 under conditions of androgen ablation. PMID- 19293290 TI - A pilot study of self-regulation informed goal setting in people with traumatic brain injury. AB - OBJECTIVE: To determine the acceptability and clinical application of two recently developed goal-setting interventions (Goal Management Training and Identity Oriented Goal Training) in people with traumatic brain injury. DESIGN: A three parallel group, randomized controlled pilot study. SETTING: Inpatient and community rehabilitation facilities. SUBJECTS: Thirty-four people with moderate to severe traumatic brain injury (Goal Management Training, n = 12; Identity Oriented Goal Training, n = 10; usual care, n = 12) and their rehabilitation clinicians. INTERVENTIONS: For both Goal Management Training and Identity Oriented Goal Training participants met face to face with their key worker weekly over a period of 6-8 weeks, during which time the key worker worked to engage them in goal setting and goal performance using the strategy prescribed by their group allocation. Usual care was provided to the other participants. MAIN MEASURES: Largely qualitative using observation, individual interviews and focus groups. Participants also completed a Goal Attainment Scale at baseline, post intervention and at three months follow-up. RESULTS: Both approaches were acceptable to the majority of participants with many reporting improved mood and goal attainment. Clinicians found working in a different way with patients both challenging and rewarding, with both experimental approaches enhancing a focus on the person's own goals. Identity Oriented Goal Training seemed particularly helpful in engaging people in the goal-setting process while Goal Management Training appeared particularly helpful in providing a structured framework for error prevention in attempting goal performance. CONCLUSION: These theoretically informed approaches to goal setting showed promise but were time intensive and at times difficult for practitioners to utilize. PMID- 19293291 TI - Identifying and applying psychological theory to setting and achieving rehabilitation goals. AB - BACKGROUND: Goal setting is considered to be a fundamental part of rehabilitation; however, theories of behaviour change relevant to goal-setting practice have not been comprehensively reviewed. OBJECTIVES: (i) To identify and discuss specific theories of behaviour change relevant to goal-setting practice in the rehabilitation setting. (ii) To identify 'candidate' theories that that offer most potential to inform clinical practice. METHODS: The rehabilitation and self-management literature was systematically searched to identify review papers or empirical studies that proposed a specific theory of behaviour change relevant to setting and/or achieving goals in a clinical context. Data from included papers were extracted under the headings of: key constructs, clinical application and empirical support. RESULTS: Twenty-four papers were included in the review which proposed a total of five theories: (i) social cognitive theory, (ii) goal setting theory, (iii) health action process approach, (iv) proactive coping theory, and (v) the self-regulatory model of illness behaviour. The first three of these theories demonstrated most potential to inform clinical practice, on the basis of their capacity to inform interventions that resulted in improved patient outcomes. CONCLUSIONS: Social cognitive theory, goal setting theory and the health action process approach are theories of behaviour change that can inform clinicians in the process of setting and achieving goals in the rehabilitation setting. Overlapping constructs within these theories have been identified, and can be applied in clinical practice through the development and evaluation of a goal-setting practice framework. PMID- 19293292 TI - The effects of an 'exercise and education' programme on exercise self-efficacy and levels of independent activity in adults with acquired neurological pathologies: an exploratory, randomized study. AB - OBJECTIVE: To evaluate the effects of an exercise and education programme on primary outcomes of exercise efficacy and activity levels; and on secondary outcomes including quality of life, mood and physical disability. DESIGN: Randomized, single blind trial. SETTING: Specialist neurosciences centre. PARTICIPANTS: Forty-four patients with acquired neurological pathologies. INTERVENTIONS: Twenty-one participants undertook a four-week exercise and education programme alongside standard follow-up care. Twenty-three participants underwent standard follow-up care alone. OUTCOME MEASURES: Primary outcome measures included the Exercise Efficacy Scale and Human Activity Profile. Secondary outcomes were evaluated by the Short Form 36 Health Survey, Hospital Anxiety and Depression Scale, UWIST Mood Adjective Checklist, Motricity Index, Frenchay Arm Test, Rivermead Mobility Index, 10-metre timed walk and a visual analogue scale. Data were collected at baseline and at 6, 12 and 24 weeks. RESULTS: At six weeks, significant improvements in exercise efficacy were found following the treatment but not the control (P= 0.001). Moderate improvements in activity levels were noted for both study groups with no between-group differences. Short Form 36 'physical health' and 'general health' domains showed trends towards increased positive change following the intervention, but these did not reach significant levels. Mood, physical impairment and physical disability remained unchanged within either study group. Longer term follow-up revealed that the improvements noted in exercise self-efficacy, activity levels and quality of life were maintained. CONCLUSIONS: The intervention resulted in improved exercise efficacy and may positively influence the physical and general health dimensions of health-related quality of life. PMID- 19293293 TI - Inner ear pathology and loss of hearing in estrogen receptor-beta deficient mice. AB - There are well known differences between males and females in hearing. In the present study, the role of estrogen receptor-beta (ER-beta; listed as ESR2 in the MGI Database) in hearing was investigated by comparing hearing and morphology of the inner ear in ER-beta knock-out mice (ER-beta(-/-)) with that of wild-type (WT) littermates. Hearing was analyzed with auditory brainstem response audiometry at 3 and 12 months. The ER-beta(-/-) mice were deaf at 1 year of age, and the morphological analysis showed absence of hair cells and loss of the whole organ of Corti initiated in the basal turn of the cochlea. Furthermore, in ER beta(-/-), but not in WT mice, the spiral ganglion was lacking many of its neurons. Immunostaining showed the presence of both ER-alpha (listed as ESR1 in the MGI Database) and ER-beta in the nuclei of some neurons in the inner ear in WT mice, but no ER-beta was found in the ER-beta(-/-) mice as expected. ER-alpha staining was predominant in the nuclei of large neurons and ER-beta in nuclei of small neurons and fibroblasts. These results reveal that both ERs are present in the inner ear at specific localizations suggesting subtype-specific functions. It is concluded that ER-beta is important for the prevention of age-related hearing loss. These findings strengthen the hypothesis that estrogen has a direct effect on hearing functions. PMID- 19293294 TI - Regulation and role of suppressor of cytokine signaling-3 in hypothalamic 4B cells. AB - Corticotropin-releasing factor (CRF) plays a central role in regulating stress responses. In the hypothalamic paraventricular nucleus (PVN), CRF, produced in response to stress, stimulates the release of ACTH from the anterior pituitary. ACTH then stimulates the release of glucocorticoids from the adrenal glands; circulating glucocorticoids are critical for recovery from stress conditions. Cytokines are also implicated in the regulation of CRF expression. Among them, interleukin (IL)-6 plays a role in the regulation of CRF. Factors other than glucocorticoids are likely to be involved in limiting the stimulation of CRF during stress. Suppressor of cytokine signaling (SOCS)-3 acts as a potent negative regulator of cytokine signaling. Little is known about the ability of the inhibitory signaling pathways to limit activation of the CRF gene in parvocellular PVN neurons. Hypothalamic 4B cells are useful for exploring the mechanisms, because these cells show characteristics of the parvocellular neurons of the PVN. In the present study, we examined whether SOCS-3 is regulated by IL-6 and cAMP in hypothalamic 4B cells. We also explored the involvement of SOCS-3 in the regulation of CRF gene expression. SOCS-3 was found to be regulated by IL-6 and via the cAMP/protein kinase A pathway in the hypothalamic cells. SOCS-3 knockdown increased IL-6- or forskolin-induced CRF gene transcription and mRNA levels. Therefore, SOCS-3, induced by a cAMP stimulant and IL-6, would be involved in the negative regulation of CRF gene expression in hypothalamic cells. PMID- 19293295 TI - Two divergent leptin paralogues in zebrafish (Danio rerio) that originate early in teleostean evolution. AB - We describe duplicate leptin genes in zebrafish (Danio rerio) that share merely 24% amino acid identity with each other and only 18% with human leptin. We were also able to retrieve a second leptin gene in medaka (Oryzias latipes). The presence of duplicate leptin genes in these two distantly related teleosts suggests that duplicate leptin genes are a common feature of teleostean fishes. Despite low primary sequence conservation, we are confident in assigning orthology between mammalian and zebrafish leptins for several reasons. First, both zebrafish leptins share their characteristic gene structure and display key features of conserved synteny with mammalian leptin genes. Secondly, the cysteine residues that make up leptin's single disulphide bridge are equally spaced in mammalian and zebrafish leptins and are unique among all members of the class-I helical cytokine family. Thirdly, the zebrafish leptins cluster with other fish leptins and mammalian leptins in phylogenetic analysis, supported by high bootstrap values. Within the leptin cluster, leptin-b forms a separate clade with the leptin-b orthologue from medaka. Finally, our prediction of the tertiary structures shows that both leptins conform to the typical four alpha-helix bundle structure of the class-I alpha-helical cytokines. The zebrafish leptins are differentially expressed; the liver shows high leptin-a expression (in concordance with what we observed for carp leptins), while leptin-b is expressed at much lower levels, which are downregulated further upon fasting. The finding of duplicate leptin genes in teleosts adds to our understanding of the evolution of leptin physiology in the early vertebrate lineage. PMID- 19293296 TI - Ageing and sexual health in the UK: how should health psychology respond to the challenges? AB - The UK has an increasing ageing population and, like other western societies, has undergone several significant shifts over the last 50 years, including attitudes towards sexual activity, relationships and growing older. Drawing on evidence from a range of sources and a number of disciplines, this editorial examines the potential sexual and reproductive health implications for current and future cohorts of ;older' people, and considers the role of health psychology in responding to the challenges of growing older in a rapidly changing (sexualized, digitalized and medicalized) world. PMID- 19293297 TI - A theory-based approach to understanding follow-up of abnormal Pap tests. AB - We applied a general theoretical framework to understand intentions to attend recommended follow-up for abnormal Pap results. Participants were 338 women attending university-affiliated clinics. Intention was associated with favorable attitudes toward follow-up (OR = 5.3); perceiving attending follow-up as consistent with one's self-concept (OR = 3.0); self-efficacy (OR = 1.8); and believing one would be told exactly what is wrong (OR = 1.3). Intention was negatively associated with believing the problem could be avoided by not returning for follow-up (OR = 0.75). Beliefs, affect and attitudes differed by race and ethnicity (all p < .05). Attendance at follow-up was related to attitude and self-concept (both p < .05). Results have implications for theory development and patient education. PMID- 19293298 TI - Assessing unrealistic optimism: impact of different approaches to measuring susceptibility to diabetes. AB - The effects of measurement method on levels of unrealistic optimism for developing diabetes were examined in 323 UK adults. A mixed factorial design comprised direct and indirect measures of unrealistic optimism as the within subjects variable, and between-subjects manipulations of the order of direct versus indirect assessment, the order of self-other comparison, the number of response options used and the polarity of the ratings. More unrealistic optimism was obtained using the direct method and using unipolar scales, and/or with the self-other order in the indirect method. The results indicate that levels of unrealistic optimism depend on its assessment method. PMID- 19293299 TI - The contribution of active and passive leisure to children's well-being. AB - The relation between leisure and well-being, including happiness and self concept, was examined in 375 children aged 8-12 years. Active leisure (e.g. physical activity) was positively correlated with well-being. Passive leisure (e.g. television and video games) was negatively correlated with well-being. Aspects of active leisure (e.g. the importance of sport to the child and how sports made the child feel) as judged by both parents and children accounted for unique variance in children's wellbeing; passive leisure did not. Similar to previous research on adolescents and adults, active leisure activities were related to children's well-being. PMID- 19293300 TI - Social support coping mediates the relationship between gender and posttraumatic growth. AB - Females tend to report greater levels of posttraumatic growth following trauma than males. Little is known about why such an association exists. This study examined whether social support coping might mediate the relationship between gender and posttraumatic growth. College students and community residing adults ( N = 221) recalled a stressful or traumatic event that they had recently experienced and responded to measures of posttraumatic growth and coping while keeping this event in mind. Gender was significantly associated with both social support coping and growth while social support coping was a partial mediator of the relationship between gender and posttraumatic growth. PMID- 19293301 TI - Objectification processes and disordered eating in British women and men. AB - The present study extended the applicability of Objectification Theory to predict disordered eating in British women and men. Participants completed measures of self-objectification, body surveillance, body shame and disordered eating. Path analyses indicated strong support for the theoretical model in women, with body shame fully mediating the relation between self-objectification and disordered eating. Patterns were similar for men with two exceptions; body shame increased with lower self-objectification and disordered eating was directly increased with higher self-objectification. Findings extend Objectification Theory as a useful framework for identifying sociocultural influences on disordered eating in British women and men. PMID- 19293302 TI - The interaction of mindful-based attention and awareness and disengagement coping with HIV/AIDS-related stigma in regard to concurrent anxiety and depressive symptoms among adults with HIV/AIDS. AB - This investigation examined the interaction of disengagement coping with HIV/AIDS related stigma and mindful-based attention and awareness in regard to anxiety and depressive symptoms among people with HIV/AIDS. There was a significant interaction in regard to anxiety symptoms. Higher levels of disengagement coping paired with lower levels of mindful-based attention and awareness was related to the greatest degrees of anxiety symptoms, while lower levels of disengagement coping paired with higher levels of mindful-based attention and awareness was related to the lowest levels of anxiety symptoms. Although the interaction for depressive symptoms was not significant, a similar pattern of results was observed. PMID- 19293303 TI - Patient and physician perceptions of the physician's explanation and patient responses to physicians. AB - Although theories on meta-cognition and self-monitoring imply the importance of meta-cognition in patient-physician interactions, there is no evidence to support this hypothesis. Thus, we evaluated patient and physician perceptions of the level of a physician's explanation and explored the possible influence of patient meta-cognition on patient responses to physicians. We conducted a questionnaire survey of 579 internist-patient pairs in Japan. The findings show that patient meta-cognition, and not perception, of the sufficiency of a physician's explanation plays a critical role in determining extreme patient responses to a physician, such as ignoring the physician's advice and doctor-shopping, whereas patient perception is a predictor of milder patient responses such as patient understanding and satisfaction. PMID- 19293304 TI - Health implications of body size perception and weight tolerance in older adults. AB - Obesity is a well-recognized risk factor for Type II diabetes and cardiovascular disease. In this study a high percentage of older adults were either overweight or obese. The aims were to assess older adults' tolerance for excess weight, and to compare self-perceptions of an acceptable weight with national guidelines. Participants were 76 males and females aged between 65 and 97 years. Results indicated that 60 per cent accurately identified their own body size. Further, participants were more tolerant of excess weight in males compared to females, regardless of age. It was concluded that this cohort remains vulnerable to weight related illnesses. PMID- 19293305 TI - 'Natural versus taught': competing discourses in antenatal breastfeeding workshops. AB - This article is an analysis of talk in breastfeeding workshops that are part of National Childbirth Trust antenatal classes. Using audio-recordings from breastfeeding workshops antenatal classes, the data were analysed using a qualitative, discursive methodology based in part on the premises outlined by Potter and Wetherell (1987) and Edwards and Potter (1992, 2001). The analysis demonstrates how there are two main discourses of breastfeeding constructed by the breastfeeding counsellor-breastfeeding as natural, and breastfeeding as learnt. In particular, it notes how these two main discourses of breastfeeding that are seemingly in competition with one another, operate concurrently within the teaching of breastfeeding, and enable the breastfeeding counsellor to manage issues and concerns around breastfeeding. PMID- 19293306 TI - Affective forecasting and advance care planning: anticipating quality of life in future health statuses. AB - That sicker people evaluate quality of life in future health status more positively, compared to healthier people, is viewed as an instance of affective forecasting error and explained by Prospect Theory, which holds that two prospects (poor health vs death) are more distinguishable when they are imminent than when distant. In a sample of 230 elderly people, we tested whether life in nine health scenarios would be more acceptable to less healthy individuals than to healthier ones. An interaction between current health status and health scenario supported the relative acceptability of poor-health prospects to sicker individuals, confirming the hypothesis. PMID- 19293307 TI - Longitudinal relations among perceived autonomy support from health care practitioners, motivation, coping strategies and dietary compliance in a sample of adults with type 2 diabetes. AB - The purpose of this study was to test the causal ordering among perceived autonomy support from health care practitioners, motivation, coping strategies and compliance to dietary self-care activities. Using a cross-lagged panel model, we investigate how these variables relate to one another over a one-year period. A total of 365 adults with Type 2 diabetes participated in the study. Results suggest that autonomous motivation and active planning are reciprocally related over time, and that prior autonomous motivation is related to the extent participants subsequently comply with their diet. Results are discussed in light of Self-determination Theory and the coping perspective. PMID- 19293308 TI - Sex disparities in cancer incidence by period and age. AB - BACKGROUND: Cancer epidemiology articles often point out that cancer rates tend to be higher among males than females yet rarely is this theme the subject of investigation. METHODS: We used the Surveillance, Epidemiology and End Results program data to compute age-adjusted (2000 U.S. standard population) sex-specific incidence rates and male-to-female incidence rate ratios (IRR) for specific cancer sites and histologies for the period 1975 to 2004. RESULTS: The 10 cancers with the largest male-to-female IRR were Kaposi sarcoma (28.73), lip (7.16), larynx (5.17), mesothelioma (4.88), hypopharynx (4.13), urinary bladder (3.92), esophagus (3.49), tonsil (3.07), oropharynx (3.06), and other urinary organs (2.92). Only 5 cancers had a higher incidence in females compared with males: breast (0.01), peritoneum, omentum, and mesentery (0.18), thyroid (0.39), gallbladder (0.57), and anus, anal canal, and anorectum (0.81). Between 1975 and 2004, the largest consistent increases in male-to-female IRR were for cancers of the tonsil, oropharynx, skin excluding basal and squamous, and esophagus, whereas the largest consistent decreases in IRR were for cancers of the lip and lung and bronchus. Male-to-female IRRs varied considerably by age, the largest increases of which were for ages 40 to 59 years for tonsil cancer and hepatocellular carcinoma. The largest decreases in male-to-female IRR by age, meanwhile, were for ages 30 to 49 years for thyroid cancer, ages >70 years for esophageal squamous cell carcinoma, and ages >30 years for lung and bronchus cancer. CONCLUSION: These observations emphasize the importance of sex in cancer etiopathogenesis and may suggest novel avenues of investigation. PMID- 19293309 TI - Association of nonsteroidal anti-inflammatory drugs with lung cancer: results from a large cohort study. AB - BACKGROUND: Lung cancer is the most common cause of cancer-related mortality. Smoking cessation is crucial to decrease risk, but additional prevention modalities are needed. The use of nonsteroidal anti-inflammatory drugs (NSAID) may be promising. METHODS: The study was a prospective cohort of 77,125 men and women, ages 50 to 76 years, from Washington state recruited in 2000 to 2002 (the VITamin And Lifestyle study). Lung cancer cases were identified through the Seattle-Puget Sound Surveillance, Epidemiology and End Results cancer registry during 5 years of follow-up. Hazard ratios (HR) associated with 10-year average use of total NSAIDs (excluding low-dose aspirin) and specific categories of NSAIDs were calculated for total incident lung cancer and specific morphologies. RESULTS: A total of 665 lung cancer cases were identified. After adjusting for smoking, age, gender, and acetaminophen use, there was a borderline-significant inverse trend with total NSAID use [>4.2 d/wk for >10 years versus none: HR, 0.82; 95% confidence interval (95% CI), 0.64-1.04; P for trend = 0.05]. The association was strongest for adenocarcinoma (HR, 0.59; 95% CI, 0.37-0.94; P for trend = 0.01) and seemed to be limited to men (HR, 0.66; 95% CI, 0.47-0.92; P for trend = 0.01) and to long-term (> or =10 years) former smokers (HR, 0.65; 95% CI, 0.44-0.96; P for trend = 0.04). There were no appreciable differences by NSAID type. CONCLUSIONS: Total NSAID use was associated with a small reduced risk of lung cancer, which was strongest for adenocarcinoma, men, and long-term former smokers. These findings are supported by known lung carcinogenesis mechanisms and suggest that NSAIDS may be useful for chemoprevention. PMID- 19293310 TI - A prospective study of relative telomere length and postmenopausal breast cancer risk. AB - During breast cancer progression, a substantial increase in chromosomal aberrations is observed in the transition from ductal hyperplasia to carcinoma in situ. Telomeres are essential structures to chromosomal integrity. Consequently, telomere dysfunction, which leads to genomic instability, is hypothesized to play a causal role in the progression of breast cancer. However, the few epidemiologic studies that have assessed the relationship between telomere length and breast cancer risk have been inconsistent. We used quantitative real-time PCR to measure relative telomere length in genomic DNA extracted from peripheral blood leukocytes and examined its association with postmenopausal breast cancer risk in 1,122 invasive breast cancer cases and 1,147 matched controls free of diagnosed cancer nested within the prospective Nurses' Health Study. Our data show that relative telomere length was not associated with a significant elevation in postmenopausal breast cancer risk [below versus above median; odds ratio, 1.23; 95% confidence interval, 0.94-1.60; P(trend) = 0.20]. Estrone and estradiol hormone levels were significantly inversely associated with relative telomere length (P = 0.02). Other established breast cancer risk factors such as family history of breast cancer and history of benign breast disease were not associated with relative telomere length in separate linear regression models each adjusted for age and disease status (P > or = 0.07). Our results provide little support for an important role of telomere length, as measured in peripheral blood leukocytes, as a biomarker of breast cancer risk. PMID- 19293311 TI - Gender is an age-specific effect modifier for papillary cancers of the thyroid gland. AB - BACKGROUND: Thyroid cancer incidence rates have increased worldwide for decades, although more for papillary carcinomas than other types and more for females than males. There are few known thyroid cancer risk factors except female gender, and the reasons for the increasing incidence and gender differences are unknown. METHODS: We used the National Cancer Institute's Surveillance, Epidemiology, and End Results 9 Registries Database for cases diagnosed during 1976-2005 to develop etiological clues regarding gender-related differences in papillary thyroid cancer incidence. Standard descriptive epidemiology was supplemented with age period-cohort (APC) models, simultaneously adjusted for age, calendar-period and birth-cohort effects. RESULTS: The papillary thyroid cancer incidence rate among females was 2.6 times that among males (9.2 versus 3.6 per 100,000 person-years, respectively), with a widening gender gap over time. Age-specific rates were higher among women than men across all age groups, and the female-to-male rate ratio declined quite consistently from more than five at ages 20-24 to 3.4 at ages 35-44 and approached one at ages 80+. APC models for papillary thyroid cancers confirmed statistically different age-specific effects among women and men (P < 0.001 for the null hypothesis of no difference by gender), adjusted for calendar-period and birth-cohort effects. CONCLUSION: Gender was an age-specific effect modifier for papillary thyroid cancer incidence. Future analytic studies attempting to identify the risk factors responsible for rising papillary thyroid cancer incidence should be designed with adequate power to assess this age specific interaction among females and males. PMID- 19293312 TI - Association of CYP1B1 haplotypes and breast cancer risk in Caucasian women. AB - CYP1B1 is a key enzyme involved in estrogen metabolism and may play an important role in the development and progression of breast cancer. In a population-based case-control study, we examined eight CYP1B1 haplotype-tagging single nucleotide polymorphisms in relation to invasive breast cancer risk. Analyses were based on 1,655 cases and 1,470 controls; all women were Caucasian. Among the individual single nucleotide polymorphisms, one (rs9341266) was associated with increased risk of breast cancer (P(trend) = 0.021), although the association was no longer significant after adjusting for multiple tests. A marginally significant haplotype effect was identified (P(global) = 0.015), with significant associations identified for 2 uncommon haplotypes comprising 4% of the controls. Results suggest that genetic variation in CYP1B1 has at most a minor influence on breast cancer susceptibility among Caucasian women. PMID- 19293313 TI - Cardiorespiratory fitness and digestive cancer mortality: findings from the aerobics center longitudinal study. AB - Although higher levels of physical activity are inversely associated with risk of colon cancer, few prospective studies have evaluated overall digestive system cancer mortality in relation to cardiorespiratory fitness (CRF). The authors examined this association among 38,801 men ages 20 to 88 years who performed a maximal treadmill exercise test at baseline in the Aerobics Center Longitudinal Study (Dallas, TX) during 1974 to 2003. Mortality was assessed over 29 years of follow-up (1974-2003). Two hundred eighty-three digestive system cancer deaths occurred during a mean 17 years of observation. Age-adjusted mortality rates per 10,000 person-years according to low, moderate, and high CRF groups were 6.8, 4.0, and 3.3 for digestive system cancer (P(trend) < 0.001). After adjustment for age, examination year, body mass index, smoking, drinking, family history of cancer, personal history of diabetes, hazard ratios (95% confidence intervals) for overall digestive cancer deaths for those in the middle and upper 40% of the distribution of CRF relative to those in the lowest 20% were 0.66 (0.49-0.88) and 0.56 (0.40-0.80), respectively. Being fit (the upper 80% of CRF) was associated with a lower risk of mortality from colon [0.61 (0.37-1.00)], colorectal [0.58 (0.37-0.92)], and liver cancer [0.28 (0.11-0.72)] compared with being unfit (the lowest 20% of CRF). These findings support a protective role of CRF against total digestive tract, colorectal, and liver cancer deaths in men. PMID- 19293314 TI - A functional genetic variant in microRNA-196a2 is associated with increased susceptibility of lung cancer in Chinese. AB - microRNAs (miRNA) are a new class of non-protein-coding, small RNAs that function as tumor suppressors or oncogenes. They participate in diverse biological pathways and function as gene regulators. Recently, we conducted a survey of common single nucleotide polymorphisms (SNP) in miRNA sequences and reported that, among four SNPs (rs2910164, rs2292832, rs11614913, and rs3746444) in pre miRNAs, rs11614913 in miR-196a2 might affect mature miR-196a expression and target mRNA-binding activity and was significantly associated with non-small cell lung cancer survival. However, it remains largely unknown whether miRNA SNPs may alter lung cancer susceptibility. In the current study, we evaluated associations between the above four SNPs in pre-miRNAs and lung cancer susceptibility in a case-control study of 1,058 incident lung cancer patients and 1,035 cancer-free controls in a Chinese population. We found that miR-196a2 rs11614913 variant homozygote CC was associated with approximately 25% significantly increased risk of lung cancer compared with their wild-type homozygote TT and heterozygote TC (odds ratio, 1.25; 95% confidence interval, 1.01-1.54). However, no significant effects were observed on the association between the other three SNPs and lung cancer risk. These findings suggest that functional SNP rs11614913 in miR-196a2 could also contribute to lung cancer susceptibility. PMID- 19293315 TI - Prenatal X-ray exposure and rhabdomyosarcoma in children: a report from the children's oncology group. AB - The association between antenatal diagnostic X-ray exposure and risk of rhabdomyosarcoma in children was assessed in a national case-control study of 319 rhabdomyosarcoma cases and 319 matched controls. Data were collected by telephone interviews of subjects' parents. Overall, an odds ratio (OR) of 1.9 [95% confidence interval (CI), 1.1-3.4] was found for any X-ray examination of the mother during pregnancy. Risk was greatest for X-ray exposure during the first trimester (OR, 5.7; 95% CI, 1.2-27.8) and was also increased for the third trimester (OR, 2.0; 95% CI, 0.9-4.6), whereas second trimester exposure was not associated with increased risk. A nonsignificant increase in risk was found for any X-rays of the abdomen, pelvis, chest, or back. Increased risk was significantly associated with "other" X-ray exposures (relative risk, 2.9; 95% CI, 1.1-7.7), primarily composed of dental X-rays. The association was strongest between embryonal rhabdomyosarcoma and first trimester exposure (relative risk, 10.5; 95% CI, 1.5-458.4). This observation regarding embryonal rhabdomyosarcoma, and our previous report of an increased frequency of major malformations in rhabdomyosarcoma are compatible with findings from animal studies in which Ptc heterozygous knockout mice exhibited an increased risk of radiation-induced development defects and of spontaneously occurring embryonal rhabdomyosarcoma. PMID- 19293316 TI - Editorial: evaluating interventions in child mental health: the importance of care provider and contextual influences. PMID- 19293317 TI - Depression in female adolescents: an IPA analysis. AB - Six female adolescents participated in open-ended interviews regarding their experiences of depression. The transcripts of their interviews were explored using Interpretative Phenomenological Analysis (Smith, 1991). The analysis revealed three inter-connected super-ordinate themes which were: communication, hurt self, and difference. These themes centred on communication processes in participants' families and with peers, issues relating to shifts in identity through adolescence, and feelings of being damaged, distressed, and hurt. Amongst a range of potentially relevant perspectives, an attachment theory (Bowlby, 1969) orientation was found to be particularly relevant and helpful in interpreting the themes that captured these young women's experiences. Some implications for clinical practice are discussed. PMID- 19293318 TI - Social capital and childhood psychiatric disorders: a cross-sectional study. AB - The aim of this research was to determine whether social capital is associated with either the type or severity of psychiatric disorders of childhood, specifically, disorders of emotion and behaviour. Ninety parents of children between the ages of 4 and 18 years with an emotional disorder or behavioural disorder participated in the study. They completed two questionnaires: the Child Behaviour Checklist (to assess severity of disorder) and a questionnaire measuring social capital. There was no statistically significant association between total social capital score and either diagnosis or severity of illness. However, two components of social capital showed significant associations: ;perceptions of the local area' was significantly associated with severity of illness and ;social networks' with diagnosis. The findings of this study suggest that some components of social capital may be more important in relation to mental health than others. PMID- 19293319 TI - Relationship factors and outcome in child anxiety treatment studies. AB - This study reviews 19 randomized controlled trials examining the association between three relationship factors - participation, treatment involvement, and therapeutic relationship - and outcome of cognitive-behavioral anxiety treatments for children and adolescents. In 12 studies, parent participation was considered as an independent variable compared to child-only participation. In three studies, parental involvement was measured. Child involvement was measured in one study. The child's perception of the therapeutic relationship was considered in three studies. Six studies found a significant positive effect of parent participation on diagnostic status, symptom level, or global functioning outcome measures. One study found a significant effect of parental involvement on global outcome measures. Another study found a significant positive association between child involvement and symptom measures and global functioning measures. No association was found between the quality of the child's perception of the therapeutic relationship and treatment outcome. Clinical implications are discussed. PMID- 19293320 TI - Therapeutic limits from an attachment perspective. AB - This article applies attachment theory and relevant research to therapeutic limit setting and focuses particularly on child-centred, non-directive play therapy (NDPT) practice. We review the role of limits in therapeutic change and examine whether therapeutic limit setting exhibits properties similar to those evident in typical adult-child relationships, a topic not previously considered in the literature. The first section identifies properties considered inherent in optimal attachment relationships from a limit setting perspective, drawing particularly on Heard and Lake's (1997) extension of Bowlby's attachment theory. The next section discusses therapists' use of limit setting from an attachment standpoint, distinguishing features of therapeutic limit setting which reflect properties evident in sensitive adult-child attachments. Finally, implications for further research and practice in child therapy are explored. PMID- 19293321 TI - Adolescents' ability to read different emotional faces relates to their history of maltreatment and type of psychopathology. AB - Emotional processing styles appear to characterize various forms of psychopathology and environmental adversity in children. For example, autistic, anxious, high- and low-emotion conduct problem children, and children who have been maltreated, all appear to show specific deficits and strengths in recognizing the facial expressions of emotions. Until now, the relationships between emotion recognition, antisocial behaviour, emotional problems, callous unemotional (CU) traits and early maltreatment have never been assessed simultaneously in one study, and the specific associations of emotion recognition to maltreatment and child characteristics are therefore unknown. We examined facial-emotion processing in a sample of 23 adolescents selected for high-risk status on the variables of interest. As expected, maltreatment and child characteristics showed unique associations. CU traits were uniquely related to impairments in fear recognition. Antisocial behaviour was uniquely associated with better fear recognition, but impaired anger recognition. Emotional problems were associated with better recognition of anger and sadness, but lower recognition of neutral faces. Maltreatment was predictive of superior recognition of fear and sadness. The findings are considered in terms of social information processing theories of psychopathology. Implications for clinical interventions are discussed. PMID- 19293322 TI - Clinical reasoning for child and adolescent mental health practitioners: the mindful formulation. AB - This article outlines a systematic approach to the formulation of clinical problems for practitioners working with children and their families. Whilst assessments in child and adolescent mental health often use a range of theoretical and practical approaches for data collection, there are relatively few resources to assist the clinician in integrating this information to develop a formulation that leads to a well considered intervention plan. The formulation approach presented here was designed to assist in training child and adolescent clinicians in a method that facilitates the process of understanding complex cases. This is done by examining the patterns of strength and difficulty identified during an assessment and systematically providing an explanation for these using ;the Four Ps' - predisposing, precipitating, perpetuating, and protective factors - to structure the clinician's thinking. Interventions that address patterns of strength and difficulty for each of the ;Four Ps' are recommended when working with complex clients. This formulation approach may take more time than merely summarizing the case, but the benefits are a more comprehensive understanding of the client. This means that problems often associated with working with complex cases can be identified and addressed, reducing the risk of drop out, poor engagement or treatment failure. PMID- 19293323 TI - Psychological needs, service utilization and provision of care in a specialist mental health clinic for young refugees: a comparative study. AB - This study addressed psychological needs, patterns of service utilization and provision of care in a specialist mental health service for young refugees and asylum seekers in London. Comparisons were made between two groups with different levels of postulated mental health need: unaccompanied minors (UAMs; n = 49) and children accompanied to the UK by one or more primary caregivers (n = 29). Significant differences were observed in referral pathways, with UAMs more likely to be referred by social services and less likely to be referred from medical agencies. UAMs also attended fewer sessions during treatment, and missed a greater proportion of scheduled appointments. Contrary to prediction, group comparisons revealed similar levels of post-migration stress and overall psychological morbidity. However, UAMs experienced significantly more traumatic events prior to resettlement, and were more likely to exhibit symptoms of post traumatic stress disorder (PTSD) than their accompanied peers. Despite their elevated risk of PTSD, UAMs were less likely than accompanied children to have received trauma-focused interventions. UAMs were also significantly less likely to have been treated using cognitive therapy, anxiety management and parent/carer training, as well as receiving fewer types of practical assistance with basic social needs. The clinical and service implications of these findings are discussed. PMID- 19293324 TI - A school-based mental health intervention for refugee children: an exploratory study. AB - This report describes an exploratory study of a school-based mental health service developed to address the psychological needs of refugee children. The service was made available in three schools and followed a consultative framework. Refugee children were discussed with the mental health team and children at greatest risk were seen. A questionnaire of psychological functioning was completed by teachers before and after the intervention. Data were collected on 47 refugee children and two control groups (ethnic minority and indigenous white children). Subgroup analyses compared children who were seen directly by the service with those for whom only consultation was provided. Refugee children had poorer overall adjustment at baseline particularly in the emotional and peer problem domains. The greatest improvements following the intervention were seen in hyperactivity for the refugee group and in peer problems for the refugees directly seen by the service. While further studies are necessary to assess its efficacy, this exploratory study indicates that an intervention which involves collaboration with teachers and parents, in an environment where children spend much of their time, can benefit vulnerable children. PMID- 19293325 TI - Test of time: what if little Albert had escaped? AB - Watson and Rayner's (1920) ;Little Albert' experiment has become one of the most famous studies in psychology. It is a staple of many general psychology textbooks and is part of the very fabric of the discipline's folklore. Despite this fame, the study has been widely criticized in the nearly 90 years since it was published for its lack of methodological rigour. This article attempts to evaluate the contribution of the ;little Albert' study to modern clinical psychology by speculating on what theories and treatments of child anxiety would look like in a parallel universe in which the study never took place because ;little Albert' escaped from the hospital in which Watson tested him. PMID- 19293326 TI - Surgical treatment of refractory tibial stress fractures in elite dancers: a case series. AB - BACKGROUND: Treatment of tibial stress fractures in elite dancers is centered on rest and activity modification. Surgical intervention in refractory cases has important implications affecting the dancers' careers. HYPOTHESIS: Refractory tibial stress fractures in dancers can be treated successfully with drilling and bone grafting or intramedullary nailing. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Between 1992 and 2006, 1757 dancers were evaluated at a dance medicine clinic; 24 dancers (1.4%) had 31 tibial stress fractures. Of that subset, 7 (29.2%) elite dancers with 8 tibial stress fractures were treated operatively with either intramedullary nailing or drilling and bone grafting. Six of the patients were followed up closely until they were able to return to dance. One patient was available only for follow-up phone interview. Data concerning their preoperative treatment regimens, operative procedures, clinical union, radiographic union, and time until return to dance were recorded and analyzed. RESULTS: The mean age of the surgical patients at the time of stress fracture was 22.6 years. The mean duration of preoperative symptoms before surgical intervention was 25.8 months. Four of the dancers were male and 3 were female. All had failed nonoperative treatment regimens. Five patients (5 tibias) underwent drilling and bone grafting of the lesion, and 2 patients (3 tibias) with completed fractures or multiple refractory stress fractures underwent intramedullary nailing. Clinical union was achieved at a mean of 6 weeks and radiographic union at 5.1 months. Return to full dance activity was at an average of 6.5 months postoperatively. CONCLUSION: Surgical intervention for tibial stress fractures in dancers who have not responded to nonoperative management allowed for resolution of symptoms and return to dancing with minimal morbidity. PMID- 19293327 TI - Wfs1 gene deletion causes growth retardation in mice and interferes with the growth hormone pathway. AB - The aim of present study was to describe changes in gene expression in the temporal lobe of mice induced by deletion of the Wfs1 gene. Temporal lobes samples were analyzed using Affymetrix Mouse Genome 420 2 GeneChips and expression profiles were functionally annotated with GSEA and Ingenuity Pathway Analysis. We found that Wfs1 mutant mice are significantly smaller (20.9 +/- 1.6 g) than their wild-type counterparts (31.0 +/- 0.6 g, P < 0.0001). This difference existed in 129S6 and C57B6 backgrounds. Interestingly, microarray analysis identified upregulation of growth hormone (GH) transcripts and functional analysis revealed activation of GH pathways. In line with microarray data, the level of IGF-1 in the plasma of Wfs1 mutant mice was significantly increased (P < 0.05). Thus, Wfs1 deletion induces growth retardation, whereas the GH pathway is activated. To test the interaction between the Wfs1 deletion and genomic background, mutant mice were backcrossed to two different genetic backgrounds. In line with previous studies, an interaction between a gene knockout and genetic background was found in gene expression profiles in the congenic region. However, genetic background did not alter the effect of the Wfs1 mutation on either body weight or GH pathway activation. Further studies are needed to describe biochemical and molecular changes of the growth hormone axis as well as in other hormones to clarify their role in growth retardation in the Wfs1 mutant mice. PMID- 19293328 TI - Characterization of a preclinical model of chronic ischemic wound. AB - Chronic ischemic wounds presenting at wound clinics are heterogeneous with respect to etiology, age of the wound, and other factors complicating wound healing. In addition, there are ethical challenges associated with collecting repeated biopsies from a patient to develop an understanding of the temporal dynamics of the mechanisms underlying chronic wounds. The need for a preclinical model of ischemic wound is therefore compelling. The porcine model is widely accepted as an excellent preclinical model for human wounds. A full-thickness bipedicle flap approach was adopted to cause skin ischemia. Closure of excisional wounds placed on ischemic tissue was severely impaired resulting in chronic wounds. Histologically, ischemic wounds suffered from impaired re epithelialization, delayed macrophage recruitment and poorer endothelial cell abundance and organization. Compared with the pair-matched nonischemic wound, unique aspects of the ischemic wound biology were examined on days 3, 7, 14, and 28 by systematic screening of the wound tissue transcriptome using high-density porcine GeneChips. Ischemia markedly potentiated the expression of arginase-1, a cytosolic enzyme that metabolizes the precursor of nitric oxide l-arginine. Ischemia also induced the SOD2 in the wound tissue perhaps as survival response of the challenged tissue. Human chronic wounds also demonstrated elevated expression of SOD2 and arginase-1. This study provides a thorough database that may serve as a valuable reference tool to develop novel hypotheses aiming to elucidate the biology of ischemic chronic wounds in a preclinical setting. PMID- 19293329 TI - New Zealand Ginger mouse: novel model that associates the tyrp1b pigmentation gene locus with regulation of lean body mass. AB - The study of spontaneous mutations in mice over the last century has been fundamental to our understanding of normal physiology and mechanisms of disease. Here we studied the phenotype and genotype of a novel mouse model we have called the New Zealand Ginger (NZG/Kgm) mouse. NZG/Kgm mice are very large, rapidly growing, ginger-colored mice with pink eyes. Breeding NZG/Kgm mice with CAST/Ei or C57BL/6J mice showed that the ginger coat colour is a recessive trait, while the excessive body weight and large body size exhibit a semidominant pattern of inheritance. Backcrossing F1 (NZG/Kgm x CAST/Ei) to NZG/Kgm mice to produce the N2 generation determined that the NZG/Kgm mouse has two recessive pigmentation variant genes (oca2(p) and tyrp-1(b)) and that the tyrp-1(b) gene locus associates with large body size. Three coat colors appeared in the N2 generation; ginger, brown, and dark. Strikingly, N2 male coat colour associated with body weight; the brown-colored mice weighed the most followed by ginger and then dark. The male brown coat-colored offspring reached adult body weights indistinguishable from NZG/Kgm males. The large NZG/Kgm mouse body size is a result of excessive lean body mass since these mice are not obese or diabetic. NZG/Kgm mice exhibit an unusual pattern of fat distribution; compared with other mouse strains they have disproportionately higher amounts of subcutaneous and gonadal fat. These mice are susceptible to high-fat diet-induced obesity but are resistant to high-fat diet-induced diabetes. We propose NZG/Kgm mice as a novel model to delineate gene(s) that regulate 1) growth and metabolism, 2) resistance to Type 2 diabetes, and 3) preferential fat deposition in the subcutaneous and gonadal areas. PMID- 19293330 TI - Genomic profiling of developing cardiomyocytes from recombinant murine embryonic stem cells reveals regulation of transcription factor clusters. AB - Cardiomyocytes derived from pluripotent embryonic stem cells (ESC) have the advantage of providing a source for standardized cell cultures. However, little is known on the regulation of the genome during differentiation of ESC to cardiomyocytes. Here, we characterize the transcriptome of the mouse ESC line CM7/1 during differentiation into beating cardiomyocytes and compare the gene expression profiles with those from primary adult murine cardiomyocytes and left ventricular myocardium. We observe that the cardiac gene expression pattern of fully differentiated CM7/1-ESC is highly similar to adult primary cardiomyocytes and murine myocardium, respectively. This finding is underlined by demonstrating pharmacological effects of catecholamines and endothelin-1 on ESC-derived cardiomyocytes. Furthermore, we monitor the temporal changes in gene expression pattern during ESC differentiation with a special focus on transcription factors involved in cardiomyocyte differentiation. Thus, CM7/1-ESC-derived cardiomyocytes are a promising new tool for functional studies of cardiomyocytes in vitro and for the analysis of the transcription factor network regulating pluripotency and differentiation to cardiomyocytes. PMID- 19293331 TI - Sulfur amino acid deficiency upregulates intestinal methionine cycle activity and suppresses epithelial growth in neonatal pigs. AB - We recently showed that the developing gut is a significant site of methionine transmethylation to homocysteine and transsulfuration to cysteine. We hypothesized that sulfur amino acid (SAA) deficiency would preferentially reduce mucosal growth and antioxidant function in neonatal pigs. Neonatal pigs were enterally fed a control or an SAA-free diet for 7 days, and then whole body methionine and cysteine kinetics were measured using an intravenous infusion of [1-(13)C;methyl-(2)H(3)]methionine and [(15)N]cysteine. Body weight gain and plasma methionine, cysteine, homocysteine, and taurine and total erythrocyte glutathione concentrations were markedly decreased (-46% to -85%) in SAA-free compared with control pigs. Whole body methionine and cysteine fluxes were reduced, yet methionine utilization for protein synthesis and methionine remethylation were relatively preserved at the expense of methionine transsulfuration, in response to SAA deficiency. Intestinal tissue concentrations of methionine and cysteine were markedly reduced and hepatic levels were maintained in SAA-free compared with control pigs. SAA deficiency increased the activity of methionine metabolic enzymes, i.e., methionine adenosyltransferase, methionine synthase, and cystathionine beta-synthase, and S-adenosylmethionine concentration in the jejunum, whereas methionine synthase activity increased and S-adenosylmethionine level decreased in the liver. Small intestine weight and protein and DNA mass were lower, whereas liver weight and DNA mass were unchanged, in SAA-free compared with control pigs. Dietary SAA deficiency induced small intestinal villus atrophy, lower goblet cell numbers, and Ki-67-positive proliferative crypt cells in association with lower tissue glutathione, especially in the jejunum. We conclude that SAA deficiency upregulates intestinal methionine cycle activity and suppresses epithelial growth in neonatal pigs. PMID- 19293333 TI - Transgenic mutant D567G but not wild-type human FSH receptor overexpression provides FSH-independent and promiscuous glycoprotein hormone Sertoli cell signaling. AB - We have characterized the in vivo actions of human wild-type FSH receptor (FSHR) overexpressed in Sertoli cells of transgenic (Tg) mice (TgFSHRwt) compared with transgenic overexpression of the human activated mutant FSHR*D567G (TgFSHR*D567G). Testicular TgFSHRwt expression significantly elevated specific FSH binding (>2-fold, P < 0.01) relative to nontransgenic testes, similar to increased FSH binding in TgFSHR*D567G testes. Isolated TgFSHRwt Sertoli cells exhibited higher FSH-stimulated cAMP levels compared with non-Tg or TgFSHR*D567G cells but did not display the elevated FSH-independent basal cAMP levels found in TgFSHR*D567G Sertoli cells. Furthermore, Sertoli cell overexpression of TgFSHR*D567G but not TgFSHRwt allowed promiscuous cAMP responses to human chorionic gonadotropin (300 IU/ml) and TSH (30 mIU/ml), demonstrating increased constitutive signaling and altered glycoprotein hormone specificity via the intracellular D567G substitution rather than FSHR overexpression. Despite elevating Sertoli cell FSH sensitivity, overexpression of TgFSHRwt had no detectable effect upon normal testis function and did not stimulate Sertoli and germ cell development in testes of gonadotropin-deficient hypogonadal (hpg) mice, in contrast to the increased meiotic and postmeiotic germ cell development in TgFSHR*D567G hpg testes. Increased steroidogenic potential of TgFSHR*D567G hpg testes was demonstrated by elevated Cyp11a1 and Star expression, which was not detected in TgFSHRwt hpg testes. Androgen-regulated and Sertoli cell-specific Rhox5 gene expression was increased in TgFSHR*D567G but not TgFSHRwt hpg testes, providing evidence of elevated LH-independent androgen activity due to mutant FSHR*D567G. Hence, transgenic FSHR overexpression in Sertoli cells revealed that the D567G mutation confers autonomous signaling and steroidogenic activity in vivo as well as promiscuous glycoprotein hormone receptor activation, independently of FSHR overexpression alone. PMID- 19293332 TI - Association of luteinizing hormone receptor gene expression with cell cycle progression in granulosa cells. AB - During hormonally induced ovarian follicle growth, granulosa cell proliferation increases and returns to baseline prior to the administration of an ovulatory stimulus. Several key genes appear to follow a similar pattern, including the luteinizing hormone receptor (LHCGR), suggesting an association between cell cycle progression and gene expression. The expression of LHCGR mRNA in granulosa cells isolated from immature rats and treated in culture with FSH increased in a time-dependent manner, whereas administration of the cell cycle inhibitor mimosine completely suppressed expression. Although forskolin was able to induce luteinization in cells treated with mimosine, human chorionic gonadotropin had no effect, indicating the functional loss of LHCGR. The effects of mimosine on cell cycle progression and LHCGR mRNA expression were reversible within 24 h of mimosine removal. Cell cycle inhibition did not alter the stability of LHCGR mRNA, indicating that the primary effect was at the transcriptional level. To determine whether the relationship between LHCGR expression and cell cycle were relevant in vivo, immature rats were given a bolus of PMSG, followed by a second injection of either saline or PMSG 24 h later to augment levels of proliferation. The expression of LHCGR mRNA was elevated in the ovaries of animals receiving a supplement of PMSG. Mimosine also blocked cell cycle progression and LHCGR mRNA expression in macaque granulosa cells isolated following controlled ovarian stimulation cycles and in two different mouse Leydig tumor lines. These data collectively indicate that LHCGR mRNA is expressed as a function of the passage of cells across the G1-S phase boundary. PMID- 19293334 TI - Role for malic enzyme, pyruvate carboxylation, and mitochondrial malate import in glucose-stimulated insulin secretion. AB - Pyruvate cycling has been implicated in glucose-stimulated insulin secretion (GSIS) from pancreatic beta-cells. The operation of some pyruvate cycling pathways is proposed to necessitate malate export from the mitochondria and NADP(+)-dependent decarboxylation of malate to pyruvate by cytosolic malic enzyme (ME1). Evidence in favor of and against a role of ME1 in GSIS has been presented by others using small interfering RNA-mediated suppression of ME1. ME1 was also proposed to account for methyl succinate-stimulated insulin secretion (MSSIS), which has been hypothesized to occur via succinate entry into the mitochondria in exchange for malate and subsequent malate conversion to pyruvate. In contrast to rat, mouse beta-cells lack ME1 activity, which was suggested to explain their lack of MSSIS. However, this hypothesis was not tested. In this report, we demonstrate that although adenoviral-mediated overexpression of ME1 greatly augments GSIS in rat insulinoma INS-1 832/13 cells, it does not restore MSSIS, nor does it significantly affect GSIS in mouse islets. The increase in GSIS following ME1 overexpression in INS-1 832/13 cells did not alter the ATP-to-ADP ratio but was accompanied by increases in malate and citrate levels. Increased malate and citrate levels were also observed after INS-1 832/13 cells were treated with the malate-permeable analog dimethyl malate. These data suggest that although ME1 overexpression augments anaplerosis and GSIS in INS-1 832/13 cells, it is not likely involved in MSSIS and GSIS in pancreatic islets. PMID- 19293335 TI - Ligand-based gene expression profiling reveals novel roles of glucocorticoid receptor in cardiac metabolism. AB - Recent studies have documented various roles of adrenal corticosteroid signaling in cardiac physiology and pathophysiology. It is known that glucocorticoids and aldosterone are able to bind glucocorticoid receptor (GR) and mineralocorticoid receptor, and these ligand-receptor interactions are redundant. It, therefore, has been impossible to delineate how these nuclear receptors couple with corticosteroid ligands and differentially regulate gene expression for operation of their distinct functions in the heart. Here, to particularly define the role of GR in cardiac muscle cells, we applied a ligand-based approach involving the GR-specific agonist cortivazol (CVZ) and the GR antagonist RU-486 and performed microarray analysis using rat neonatal cardiomyocytes. We indicated that glucocorticoids appear to be a major determinant of GR-mediated gene expression when compared with aldosterone. Moreover, expression profiles of these genes highlighted numerous roles of glucocorticoids in various aspects of cardiac physiology. At first, we identified that glucocorticoids, via GR, induce mRNA and protein expression of a transcription factor Kruppel-like factor 15 and its downstream target genes, including branched-chain aminotransferase 2, a key enzyme for amino acid catabolism in the muscle. CVZ treatment or overexpression of KLF15 decreased cellular branched-chain amino acid concentrations and introduction of small-interfering RNA against KLF15 cancelled these CVZ actions in cardiomyocytes. Second, glucocorticoid-GR signaling promoted gene expression of the enzymes involved in the prostaglandin biosynthesis, including cyclooxygenase-2 and phospholipase A2 in cardiomyocytes. Together, we may conclude that GR signaling should have distinct roles for maintenance of cardiac function, for example, in amino acid catabolism and prostaglandin biosynthesis in the heart. PMID- 19293337 TI - A mutation linked to retinitis pigmentosa in HPRP31 causes protein instability and impairs its interactions with spliceosomal snRNPs. AB - The AD29 mutation in HPRP31 belongs to a series of mutations that were initially linked with the autosomal dominant disorder retinitis pigmentosa (RP) type 11. The HPRP31 gene encodes the hPrp31 protein that specifically associates with spliceosomal small nuclear ribonucleoprotein particles (snRNPs). Despite intensive research, it is still unclear how the AD29 (Ala216Pro) mutation causes RP. In this study, we report that the expression of this mutant protein affects cell proliferation and alters the structure of nuclear Cajal bodies that are connected with snRNP metabolism. Interestingly, these effects can be reversed by the over-expression of the hPrp6 protein, a binding partner of hPrp31. Although Ala216 is not contained within the U4 or U5 snRNP interacting domains, we present several lines of evidence that demonstrate that the association between the AD29 mutant and snRNPs in the cell nucleus is significantly reduced. Finally, we show that the stability of the AD29 mutant is severely affected resulting in its rapid degradation. Taken together, our results indicate that the Ala216Pro mutation destabilizes the hPrp31 protein structure in turn reducing its interaction with snRNP binding partners and leading to its rapid degradation. These findings significantly impact our understanding of the molecular mechanisms underlying RP and suggest that the insufficiency of the functional hPrp31 protein combined with the potential cytotoxicity associated with the expression the AD29 mutant are at least partially causative of the RP phenotype. PMID- 19293336 TI - Regulated renin release from 3T3-L1 adipocytes. AB - Whereas adipose tissue possesses a local renin-angiotensin system, the synthesis and regulated release of renin has not been addressed. To that end, we utilized differentiating 3T3-L1 cells and analyzed renin expression and secretion. Renin mRNA expression and protein enzymatic activity were not detectable in preadipocytes. However, upon differentiation, renin mRNA and both intracellular and extracellular renin activity were upregulated. In differentiated adipocytes, forskolin treatment resulted in a 28-fold increase in renin mRNA, whereas TNFalpha treatment decreased renin mRNA fourfold. IL-6, insulin, and angiotensin (Ang) II were without effect. In contrast, forskolin and TNFalpha each increased renin protein secretion 12- and sevenfold, respectively. Although both forskolin and TNFalpha induce lipolysis in adipocytes, fatty acids, prostaglandin E(2), and lipopolysaccharide had no effect on renin mRNA or secretion. To evaluate the mechanism(s) by which forskolin and/or TNFalpha are able to regulate renin secretion, a general lipase inhibitor (E600) and PKA inhibitor (H89) were used. Both inhibitors attenuated forskolin-induced renin release, whereas they had no effect on TNFalpha-regulated secretion. In contrast, E600 potentiated forskolin stimulated renin mRNA levels, whereas H89 had no effect. Neither inhibitor had any influence on TNFalpha regulation of renin mRNA. Relative to lean controls, renin expression was reduced 78% in the epididymal adipose tissue of obese male C57Bl/6J mice, consistent with TNFalpha-mediated downregulation of renin mRNA in the culture system. In conclusion, the expression and secretion of renin are regulated under a complex series of hormonal and metabolic determinants in mature 3T3-L1 adipocytes. PMID- 19293338 TI - Molecular mechanisms for subtelomeric rearrangements associated with the 9q34.3 microdeletion syndrome. AB - We characterized at the molecular level the genomic rearrangements in 28 unrelated patients with 9q34.3 subtelomeric deletions. Four distinct categories were delineated: terminal deletions, interstitial deletions, derivative chromosomes and complex rearrangements; each results in haploinsufficiency of the EHMT1 gene and a characteristic phenotype. Interestingly, 25% of our patients had de novo interstitial deletions, 25% were found with derivative chromosomes and complex rearrangements and only 50% were bona fide terminal deletions. In contrast to genomic disorders that are often associated with recurrent rearrangements, breakpoints involving the 9q34.3 subtelomere region are highly variable. Molecular studies identified three regions of breakpoint grouping. Interspersed repetitive elements such as Alu, LINE, long-terminal repeats and simple tandem repeats are frequently observed at the breakpoints. Such repetitive elements may play an important role by providing substrates with a specific DNA secondary structure that stabilizes broken chromosomes or assist in either DNA double-strand break repair or repair of single double-strand DNA ends generated by collapsed forks. Sequence analyses of the breakpoint junctions suggest that subtelomeric deletions can be stabilized by both homologous and nonhomologous recombination mechanisms, through a telomere-capture event, by de novo telomere synthesis, or multistep breakage-fusion-bridge cycles. PMID- 19293339 TI - Genetic ablation of cyclophilin D rescues mitochondrial defects and prevents muscle apoptosis in collagen VI myopathic mice. AB - Ullrich congenital muscular dystrophy (UCMD) and Bethlem myopathy are inherited muscle disorders caused by mutations of genes encoding the extracellular matrix protein collagen VI (ColVI). Mice lacking ColVI (Col6a1(-/-)) display a myopathic phenotype associated with ultrastructural alterations of mitochondria and sarcoplasmic reticulum, mitochondrial dysfunction with abnormal opening of the permeability transition pore (PTP) and increased apoptosis of muscle fibers. Treatment with cyclosporin (Cs) A, a drug that desensitizes the PTP by binding to cyclophilin (Cyp)-D, was shown to rescue myofiber alterations in Col6a1(-/-) mice and in UCMD patients, suggesting a correlation between PTP opening and pathogenesis of ColVI muscular dystrophies. Here, we show that inactivation of the gene encoding for Cyp-D rescues the disease phenotype of ColVI deficiency. In the absence of Cyp-D, Col6a1(-/-) mice show negligible myofiber degeneration, rescue from mitochondrial dysfunction and ultrastructural defects, and normalized incidence of apoptosis. These findings (i) demonstrate that lack of Cyp-D is equivalent to its inhibition with CsA at curing the mouse dystrophic phenotype; (ii) establish a cause-effect relationship between Cyp-D-dependent PTP regulation and pathogenesis of the ColVI muscular dystrophy and (iii) validate Cyp-D and the PTP as pharmacological targets for the therapy of human ColVI myopathies. PMID- 19293340 TI - Reproductive and epigenetic outcomes associated with aging mouse oocytes. AB - Female aging entails a decline in fertility in mammals, manifested by reduced oocyte reserves and poor oocyte quality accompanied by chromosomal anomalies and reduced litter size. In addition to compromised genetic integrity, recent studies suggest that epigenetic mechanisms may be altered in aging oocytes, with age affecting the expression of DNA methyltransferases, which catalyze the important epigenetic modification, DNA methylation. Loss of DNA methylation patterns, most notably for imprinted genes, is lethal to mouse embryos. To investigate how maternal age affects embryonic development and underlying DNA methylation patterns, young and aged C57BL/6 females were mated with C57BL/6 or C57BL/6(CAST7) males to allow for the identification of parental alleles; resulting blastocysts and mid-gestation embryos and placentas were evaluated. Although pregnancy, ovulation and implantation rates were similar between age groups, an age-related increase in resorption sites, morphological abnormalities and delayed development was found. Interestingly, placental morphology was also perturbed by aging, with elevated numbers of trophoblast giant cells in aged pregnancies. Normal monoallelic expression of the imprinted genes H19 and Snrpn was unaltered in blastocysts from aged females. We failed to observe any age related changes in methylation of the differentially methylated regions of imprinted genes Snrpn, Kcnq1ot1, U2af1-rs1, Peg1, Igf2r and H19. Restriction Landmark Genome Scanning showed no significant differences in genome-wide DNA methylation in embryos and placentas, regardless of maternal age. Our findings demonstrate that maternal age affects post-implantation embryo and placental development; however embryos capable of developing to mid-gestation appear to undergo normal acquisition and maintenance of DNA methylation patterning. PMID- 19293341 TI - MicroRNAs and genomic variations: from Proteus tricks to Prometheus gift. AB - MicroRNAs (miRNAs) are small non-coding RNAs with regulatory functions. MiRNAs are aberrantly expressed in almost all human cancers, leading to abnormal levels of target genes. Recently, an increasing number of studies have addressed whether genomic variations including germ line or somatic mutations and single-nucleotide polymorphisms can count for miRNA abnormal expression by altering their biogenesis and/or affect the ability of miRNAs to bind to target messenger RNAs. Here, we provide an extensive review of the studies that have investigated variations occurring both in miRNA genes and in target genes and we discuss the possible clinical implications of these findings. Furthermore, we propose that sequence variations in miRNAs or interactor sites located in mRNAs can be involved in cancer predisposition. PMID- 19293342 TI - Effect of food on the pharmacokinetics and pharmacodynamics of an oral ghrelin agonist (ARD-07) in healthy subjects. AB - ARD-07 (also known as EP01572) is a peptidomimetic growth hormone secretagogue that can be administered orally. The primary objective of this study is to determine the effects of a meal on the oral bioavailability of ARD-07 after a single oral dose (0.5 mg/kg). In addition, the pharmacodynamic effects (growth hormone release, insulin-like growth factor-1 concentrations) and the tolerability of ARD-07 are investigated in this open-label, randomized, crossover study. Sixteen healthy subjects (8 males, 8 females) receive ARD-07 on 2 different days; the treatment consists of a single oral dose of ARD-07 (0.5 mg/kg body weight), once with and the second day without a test meal. Plasma kinetics of ARD-07 and pharmacodynamic effects are quantified by specific assays. Results are given as mean +/- SEM: The area under the curve for 0 to 24 hours is approximately twice as high without food (27.8 +/- 4.1) than with food (13.7 +/- 1.2; P = .002). The maximum observed ARD-07 concentration relative to dose administration (C(max)) is more than twice as high without food (10.6 +/- 1.6 ng/mL) than with food (4.4 +/- 0.5 ng/mL; P = .001). C(max) of growth hormone occurs at a significantly (P = .001) later stage with food (C(max) = 13.0 +/- 3.5 ng/mL) than without food (37.1 +/- 5.3 ng/mL). Food has a marked effect on the absorption of ARD-07: there is a significant difference in bioavailability between administration of oral ARD-07 with and without food. PMID- 19293344 TI - Search and rescue: finding ways to correct deltaF508 CFTR. PMID- 19293347 TI - Applying translational research in understanding complications and defining targets for intervention: inflammation in PD as a model. AB - The bench-to-bedside approach to translational research is becoming increasingly important to efficiently advance understanding of the mechanisms underlying disease and to improve the quality of patient care. Although this investigation model has been practiced since the early days of the therapy, robust research platforms built to practice translational research have only recently been structured in the field of peritoneal dialysis. Experience with a translational research environment that generated most of the information cited in this overview is the core of this manuscript. The central investigation theme described is how to approach the cardiovascular complications of peritoneal dialysis. The research question was, could the continuous activation of inflammatory pathways be central in this process and represent a relevant target for interventions? PMID- 19293348 TI - Absolute free choice for dialysis modality selection -- is it possible? PMID- 19293349 TI - Increasing PD utilization: should suitable patients be forced? PMID- 19293350 TI - Biocompatibility and peritoneal transport. PMID- 19293351 TI - Previous comorbidity and lack of patient free choice of technique predict early mortality in peritoneal dialysis. AB - OBJECTIVE: To study the prognostic factors for mortality and hospital admission for patients on peritoneal dialysis (PD). METHOD: Biannual data on individual characteristics, clinical and analytical progress, treatment, and events were studied for a cohort of incident patients undergoing PD (2003-2006) in a reference area of 8.8 million people. RESULTS: 489 patients (age 53.58 years, 61.6% male) with 3-year follow-up were included. They presented at inclusion with Charlson Comorbidity Index (CCI) of 5.25; previous cardiovascular (CV) event, 23.7%; diabetes mellitus (DM), 19.1%; and hypertension (HT), 89.9%. Annual hospitalization rate per patient-year at risk was 0.6. The variables that predicted admission were CCI [odds ratio (OR) 1.14 per point], DM (OR 1.66), and previous CV event (OR 1.90). Anemia maintained significance when corrected for CCI: hemoglobin, 0.79 per 1 g/dL Hb; CCI, 1.15 per point. Annual mortality rate was 5.4%. Those that died were older (67.47 vs 52.78 years) and had a higher CCI (8.35 vs 5.0), a lower initial Hb (11.5 vs 12.2 g/dL), a higher hospital admission rate, a higher annual rate of peritonitis, more previous CV events (50.0% vs 22.1%), and higher prevalence of DM (38.5% vs 17.9%). Survival analysis identified the following prognostic factors: CCI [hazard ratio (HR) 1.51 per point], CV event (HR 2.85), DM (HR 2.52), age (HR 1.06 per year), and mandatory referral to PD (HR 6.54). The effect of CV events and DM persisted after correction for age, and that of choice of technique after correcting for CCI and/or age. CONCLUSIONS: The CCI is useful for risk estimation in PD patients. Previous CV event, DM, and age are the most relevant risk factors. Control of anemia has prognostic value for hospital admissions. Mandatory referral to PD is associated with higher mortality. The prognosis in PD depends on predialysis patient management. PMID- 19293352 TI - Effects of ionized sodium concentrations on ultrafiltration rate in peritoneal dialysis using lactate and lactate/bicarbonate solutions. AB - OBJECTIVE: To investigate the possible effects of different concentrations of ionized sodium (NaI) on peritoneal ultrafiltration (UF) rate using lactate (Lac) and lactate/bicarbonate (Lac/Bic) dialysis solutions. DESIGN: Two random consecutive (after an interval of 48 hours) peritoneal equilibration tests (PETs) were performed in 13 patients (4 males and 9 females) on regular continuous ambulatory peritoneal dialysis (PD) treatment for at least 3 months. Two different PD solutions containing anhydrous glucose 3.86% were used: a 40 mmol/L Lac solution and a 15/25 mmol/L mixed Lac/Bic solution. Concentrations of total sodium (NaT) and NaI were measured by flame photometer and direct ion-selective electrode respectively. RESULTS: Dialysate concentrations of NaT were not different during PETs using Lac and Lac/Bic. Dialysate concentrations of NaI in fresh PD solutions were different (133.3 +/- 1.7 vs 128.2 +/- 3.9 mmol, p < 0.0001); however, these differences disappeared just after the end of the infusion of the fresh solutions. Peritoneal UF rate was not significantly different during PETs using Lac versus Lac/Bic (609 +/- 301 mL vs 542 +/- 362 mL). The dialysate-to-plasma ratios of sodium concentrations at 60 minutes of the PETs (which are expressions of free water transport) were not different using Lac versus Lac/Bic (0.89 +/- 0.04 vs 0.89 +/- 0.04 respectively, p = 0.96). All the other classical parameters of the PET were not different between Lac and Lac/Bic. CONCLUSIONS: The higher dialysate concentrations of NaI due to lower dialysate pH and consequently the higher effective osmolality of the fresh Lac PD solutions did not influence peritoneal UF rate, probably because of the fast reduction of NaI concentrations due to rapid correction of dialysate pH at the end of the infusion of Lac solutions into the peritoneal cavity. PMID- 19293353 TI - Association between pulse pressure and mortality in patients undergoing peritoneal dialysis. AB - BACKGROUND: Pulse pressure has been shown to be associated with adverse outcomes in the general population and in patients on hemodialysis (HD). However, the significance of pulse pressure has not been studied in peritoneal dialysis (PD) patients. This study examined the association between pulse pressure and mortality in patients undergoing chronic PD. METHODS: All patients aged 18 years or older that commenced PD between 1 January 2000 and 31 December 2005 at the University Health Network, Toronto, were included. The association between pulse pressure and mortality was assessed using the Cox proportional hazards model. RESULTS: A total of 306 patients were included in the study. Mean pulse pressure of the study cohort was 56.8 +/- 17.8 mmHg. Age and diabetes were significant predictors of elevated pulse pressure (p < 0.001). After adjusting for the level of systolic blood pressure and other demographic and clinical parameters, multivariable Cox proportional hazards modeling showed a direct and consistent association between pulse pressure and death risk. Each increment of 1 mmHg in pulse pressure was associated with a 2.7% increased hazard of all-cause death [95% confidence interval (CI) 1.001 - 1.054, p = 0.039] and a 4.1% increase in risk for cardiovascular mortality (hazard ratio 1.041, 95% CI 1.003 - 1.081; p = 0.035). CONCLUSION: Elevated pulse pressure is associated with an increased risk of all-cause and cardiovascular death in patients on PD. Recognition of this characteristic as an important predictor of mortality suggests that one goal of antihypertensive therapy in PD patients should be to decrease elevated pulse pressure. PMID- 19293354 TI - Oxidative stress in children on peritoneal dialysis. AB - OBJECTIVES: Enhanced oxidative stress has been observed in dialysis and predialysis adult patients with chronic kidney disease (CKD), which resulted in increased mortality and morbidity within this population. Not much attention in the literature has been paid to nonenzymatic antioxidant defense in children with CKD on peritoneal dialysis (PD). The aim of the present study was to describe the plasma, erythrocyte, and dialysate concentrations of oxidized (GSSG) and reduced glutathione (GSH) and vitamins A, E, and C in a pediatric PD population. PATIENTS: 10 children on PD and 27 age-matched healthy subjects were enrolled in the study. RESULTS: Erythrocyte and plasma GSH concentrations were lower in PD patients, erythrocyte concentration of GSSG remained unchanged, and plasma GSSG was significantly higher in children on PD. Children on PD exhibited decreased plasma concentrations of antioxidant vitamins compared to healthy subjects. Moreover, we documented loss of vitamins A, E, and C into ultrafiltrate. CONCLUSION: Such low plasma levels of vitamins A, E, and C and simultaneously decreased activity of erythrocyte GSH may be responsible for the increased oxidative stress occurring in children with CKD on PD. PMID- 19293355 TI - Effects of beraprost sodium, an oral prostaglandin i2 analog, on hemostatic factors and inflammation in chronic peritoneal dialysis patients. AB - Beraprost sodium, an orally active prostaglandin I2 analog with vasodilatory, cytoprotective, antiplatelet, antithrombotic, and anti-inflammatory effects, 120 microg daily for 8 weeks, decreased plasma D-dimer, a marker of intravascular coagulation, and von Willebrand factor, a marker for endothelial injury, in 100 chronic peritoneal dialysis patients. Total cholesterol, triglycerides, high density lipoprotein, apolipoprotein A1, apolipoprotein B, albumin, prealbumin, fibrinogen, troponin-T, and high-sensitivity C-reactive protein levels were not changed. Three patients complained of headache and 1 patient experienced facial flushing; however, no serious adverse effects were observed. These results suggest that beraprost sodium is effective in partially reversing the thrombogenic coagulation profile and endothelial injury in chronic peritoneal dialysis patients. PMID- 19293356 TI - Long-term modality-related mortality analysis in incident dialysis patients. AB - BACKGROUND: The effects of the various dialysis modalities on patient survival are different, especially for diabetic patients. Hemodialysis (HD) and peritoneal dialysis (PD) are the predominant renal replacement modalities. This study analyzes modality-related mortality in long-term dialysis patients. METHODS: This prospective cohort study was conducted between May 1991 and October 2005. Incident patients that had initiated dialysis and had been on dialysis for more than 3 months were enrolled. All cause, infection related, and cardiovascular disease-related mortalities were used as end points. Patient survival was analyzed by the Cox proportional hazards model after adjusting for age, sex, diabetes, comorbidity, and time-averaged values of laboratory data to control influential covariates. RESULTS: In total, 1347 patients (258 on PD and 1089 on HD) were enrolled. Adjusted all cause, infection related, and cardiovascular disease-related mortality did not differ significantly between HD and PD patients. In diabetic patients, adjusted all-cause [HD vs PD: hazard ratio (HR) 0.717, 95% confidence interval (CI) 0.400 - 1.282] and infection-related mortality (HD vs PD: HR 1.341, 95% CI 0.453 - 3.969) did not differ significantly between patients on HD and patients on PD. However, adjusted cardiovascular disease-related mortality increased significantly in diabetic PD patients (HD vs PD: HR 0.375, 95% CI 0.154 - 0.913). For nondiabetic patients, adjusted all cause, infection related, and cardiovascular disease-related mortality did not differ significantly between HD and PD patients. CONCLUSIONS: Dialysis modality had no significant impact on all-cause or infection-related mortality. More studies are needed to clarify the putative difference in cardiovascular mortality risk between diabetic patients on PD and diabetic patients on HD. PMID- 19293357 TI - Pulmonary hypertension in peritoneal dialysis patients: prevalence and risk factors. AB - AIM: To investigate the prevalence of pulmonary arterial hypertension (PAH) and the possible contributing factors for PAH in patients receiving regular continuous ambulatory peritoneal dialysis (CAPD). PATIENTS AND METHODS: The study included 135 CAPD patients and 15 disease-free controls. Patients that had chronic obstructive pulmonary disease, severe mitral or aortic valve disease, connective tissue disease, history of pulmonary embolism, left ventricular ejection fraction <50%, or chest wall or parenchymal lung disease were excluded. All patients and controls were examined using echocardiography and bioelectrical impedance analysis. PAH was defined as systolic pulmonary artery pressure (PAP) >35 mmHg at rest. RESULTS: Mean systolic PAP was higher in the CAPD patients than in the controls (19.66 +/- 11.66 vs 14.27 +/- 4.55 mmHg, p = 0.001). PAH was detected in 17 (12.6%) of the 135 CAPD patients. Mean systolic PAP was significantly higher in patients with PAH than in those without PAH (42.00 +/- 9.13 vs 16.44 +/- 7.83 mmHg, p = 0.001). Serum albumin level and ejection fraction were lower in patients with PAH than in those without PAH (p = 0.001 and 0.003 respectively). The ratio of extracellular water/total body water (ECW/TBW), which can reflect hydration status, was significantly higher in patients with PAH than in those without PAH (p = 0.008). In the PD group, no patients were hypovolemic; 51 (37.8%) of the 135 PD patients were hypervolemic and 84 (62.2%) were normovolemic. Only 3 of the 17 patients with PAH were normovolemic; the rest were hypervolemic. Mean systolic PAP was significantly higher in hypervolemic PD patients (24.57 +/- 14.19 mmHg) than in normovolemic PD patients (16.68 +/- 7.61 mmHg) (p = 0.001). PAP correlated with ECW/TBW (r = 0.317, p = 0.001) and left ventricular mass index (LVMI; r = 0.286, p = 0.001). On the other hand, it inversely correlated with serum albumin level (r = -0.281, p = 0.001), hemoglobin level (r = -0.165, p = 0.044), and ejection fraction (r = -0.263, p = 0.001). Serum albumin level, ECW/TBW, and LVMI were found in multivariate analysis to be independent risk factors for PAP. CONCLUSION: PAH is a frequent cardiovascular complication in CAPD patients. Serum albumin level, hypervolemia, and LVMI are major risk factors for PAH. Therefore, strategies for treatment of hypervolemia, left ventricular hypertrophy, and hypoalbuminemia should be enhanced to prevent the development of PAH in CAPD patients. PMID- 19293358 TI - Subcutaneous administration of darbepoetin alfa effectively maintains hemoglobin concentrations at extended dose intervals in peritoneal dialysis patients. AB - OBJECTIVE: Darbepoetin alfa is an erythropoietic-stimulating protein with a threefold longer half-life than recombinant human erythropoietin (rHuEPO) and can be used less frequently in the treatment of renal anemia. The purpose of this single-center single-arm study was to determine whether darbepoetin alfa, when administered at extended dose intervals, is as effective as rHuEPO for the treatment of renal anemia in patients on peritoneal dialysis. METHODS: Patients on peritoneal dialysis for at least 3 years receiving stable rHuEPO therapy were shifted to darbepoetin alfa administered every week, or every other week, using the recommended 200:1 conversion factor. The doses of darbepoetin alfa were titrated to maintain hemoglobin within +/-1.0 g/dL of patients' baseline values and within a range of 9.0 - 12.0 g/dL for up to 24 weeks (20-week dose titration period followed by 4-week evaluation period). The primary end point was the change in hemoglobin levels between baseline and evaluation period. RESULTS: 73 patients completed the study; mean age was 52.1 years; 30 males. Mean baseline and evaluation period hemoglobin levels were similar (9.56 +/- 1.11 vs 9.73 +/- 1.41 g/dL, p = 0.248). Mean rHuEPO dose was 92.9 IU/kg/week (equivalent to 0.46 microg/kg/week darbepoetin alfa), which was higher than darbepoetin alfa dose during the evaluation period (0.46 vs 0.34 microg/kg/week, p = 0.038). In addition, ferritin levels decreased (483 +/- 26 vs 396 +/- 19 ng/dL, p = 0.014). The other parameters, such as albumin, C-reactive protein, transferrin saturation, Kt/V, and weekly creatinine clearance showed no statistical difference between the two regimens. No serious or major adverse effects were observed with darbepoetin alfa during the study. CONCLUSIONS: Using lower dosage and frequency, darbepoetin alfa effectively maintains hemoglobin levels in peritoneal dialysis patients previously maintained on erythropoietin beta. Similar effects on hemoglobin can be maintained with even lower levels of ferritin during darbepoetin alfa use. These results show that darbepoetin alfa is safe, effective, and convenient in treating renal anemia in peritoneal dialysis patients. PMID- 19293359 TI - Evaluation of taurine as an osmotic agent for peritoneal dialysis solution. AB - OBJECTIVE: The development of a glucose-free peritoneal dialysis (PD) solution is important because glucose has been associated with functional and morphological damage to the peritoneal membrane. The ultrafiltration (UF) and biocompatibility of new PD solutions containing taurine (PD-taurine) instead of glucose as an osmolite were tested in a rat PD model. METHODS: To determine the solution's UF ability, different concentrations of taurine in PD solutions were compared to glucose-based PD solutions (PD-glucose) by giving single intraperitoneal injections for 2, 4, and 6 hours. To examine the biocompatibility of PD-taurine, the rats were divided into 3 groups: a 3.86% PD-glucose group, a 3.5% PD-taurine group and a not dialyzed group. The rats were given 10-mL injections of PD fluids intraperitoneally 3 times daily for 7 days. A peritoneal equilibration test (PET) was performed using a 1.9% xylitol solution at the time the rats were sacrificed. Mesothelial cell monolayers were obtained from the animals and studied based on a population analysis. RESULTS: The net UF of PD-taurine increased in a dose dependent manner; the 3.5% PD-taurine solution was equivalent to the 3.86% PD glucose solution after 4 hours. The PET showed that the drainage volume and the D(4)/D(0) ratio for xylitol after 4 hours with PD-taurine solution were significantly greater than with the PD-glucose solution (p < 0.001 and p < 0.001 respectively). Mesothelial and fibroblast-like cell proliferation was significantly less with PD-taurine than with PD-glucose (p < 0.01). CONCLUSIONS: These results indicate that PD-taurine resulted in net UF equivalent to that of PD-glucose and was more biocompatible than PD-glucose with respect to the peritoneal membrane. PMID- 19293360 TI - HPMCs induce greater intercellular delocalization of tight junction-associated proteins due to a higher susceptibility to H2O2 compared with HUVECs. AB - BACKGROUND: Reactive oxygen species (ROS) have been speculated as possible inducers of structural or functional changes that lead to a hyperpermeable state in patients on long-term peritoneal dialysis. This study aimed to compare localization of tight junction-associated proteins (TJPs), which relate to solute permeability characteristics, between human peritoneal mesothelial cell (HPMC) monolayers and human umbilical vein endothelial cell (HUVEC) monolayers under oxidative stress. METHODS: HPMCs and HUVECs were cultured on a polymer mesh until transepithelial electrical resistance reached a plateau. Solute permeation tests were conducted using FITC-labeled dextrans. Localization of TJPs was observed under a confocal laser scanning microscope. These experiments were carried out with/without 0.1 mmol/L H(2)O(2). In addition, ROS production as well as the amounts of intracellular reductive glutathione (GSH) and oxidative glutathione were measured. RESULTS: When the monolayers were exposed to 0.1 mmol/L H(2)O(2)/medium for 2 hours, the HPMC monolayer revealed a significant reduction in transepithelial electrical resistance (from 32.5 +/- 3.4 to 17.4 +/- 4.9 Omega.cm(2)) with delocalization of TJPs, particularly occludins. The HUVEC monolayer remained stable and exhibited an unremarkable change in TJP organization. Compared to the HUVEC monolayer, the HPMC monolayer exhibited two- to threefold higher 2',7'-dichlorofluorescein intensities that increased in a dose-dependent manner. HUVECs contained approximately 2.5-times more GSH than HPMCs. This supported the lesser production of ROS when exposed to 0.1 mmol/L H(2)O(2) for 24 hours. HUVECs used 8.03 nmol/mg GSH protein to maintain TJP localization, while only 3.75 nmol/mg GSH protein was available for the HPMCs. CONCLUSION: The HUVEC monolayer, which was less permeable to middle-to-high molecular weight solutes, was more tolerant against ROS stress than the HPMC monolayer. Availability of intracellular GSH is an important issue in maintaining the integrity of the mesothelium. PMID- 19293361 TI - Chronic exposure of mouse peritoneum to peritoneal dialysis fluid: structural and functional alterations of the peritoneal membrane. PMID- 19293362 TI - Evolution of gram-negative bacilli susceptibility in peritoneal dialysis-related peritonitis in Brazil: a single center's experience over nine years. PMID- 19293363 TI - Adverse effects of pantoea peritonitis on peritoneal transport. PMID- 19293364 TI - Alternaria peritonitis after contact with a cat. PMID- 19293365 TI - Hemoperitoneum: a problem in PD patients with hepatic tumors. PMID- 19293366 TI - Polymicrobial CAPD peritonitis after hysteroscopy. PMID- 19293367 TI - CAPD assisted by family member -- a single-center experience. PMID- 19293368 TI - Uridine ribohydrolase and the balance between nucleotide degradation and salvage. PMID- 19293369 TI - The gene controlling the indole glucosinolate modifier1 quantitative trait locus alters indole glucosinolate structures and aphid resistance in Arabidopsis. AB - Glucosinolates are defensive secondary compounds that display large structural diversity in Arabidopsis thaliana and related plants. Much attention has been paid to variation in the biosynthesis of Met-derived aliphatic glucosinolates and its ecological consequences, but little is known about the genes that cause qualitative and quantitative differences in Trp-derived indole glucosinolates. We use a combination of quantitative trait locus (QTL) fine-mapping and microarray based transcript profiling to identify CYP81F2 (At5g57220), encoding a cytochrome P450 monooxygenase, as the gene underlying Indole Glucosinolate Modifier1 (IGM1), a metabolic QTL for the accumulation of two modified indole glucosinolates, 4 hydroxy-indole-3-yl-methyl and 4-methoxy-indole-3-yl-methyl glucosinolate. We verify CYP81F2 function with two SALK T-DNA insertion lines and show that CYP81F2 catalyzes the conversion of indole-3-yl-methyl to 4-hydroxy-indole-3-yl-methyl glucosinolate. We further show that the IGM1 QTL is largely caused by differences in CYP81F2 expression, which results from a combination of cis- and trans-acting expression QTL different from known regulators of indole glucosinolate biosynthesis. Finally, we elucidate a potential function of CYP81F2 in plant insect interactions and find that CYP81F2 contributes to defense against the green peach aphid (Myzus persicae) but not to resistance against herbivory by larvae from four lepidopteran species. PMID- 19293370 TI - Uridine-ribohydrolase is a key regulator in the uridine degradation pathway of Arabidopsis. AB - Nucleoside degradation and salvage are important metabolic pathways but hardly understood in plants. Recent work on human pathogenic protozoans like Leishmania and Trypanosoma substantiates an essential function of nucleosidase activity. Plant nucleosidases are related to those from protozoans and connect the pathways of nucleoside degradation and salvage. Here, we describe the cloning of such an enzyme from Arabidopsis thaliana, Uridine-Ribohydrolase 1 (URH1) and the characterization by complementation of a yeast mutant. Furthermore, URH1 was synthesized as a recombinant protein in Escherichia coli. The pure recombinant protein exhibited highest hydrolase activity for uridine, followed by inosine and adenosine, the corresponding K(m) values were 0.8, 1.4, and 0.7 mM, respectively. In addition, URH1 was able to cleave the cytokinin derivative isopentenyladenine riboside. Promoter beta-glucuronidase fusion studies revealed that URH1 is mainly transcribed in the vascular cells of roots and in root tips, guard cells, and pollen. Mutants expressing the Arabidopsis enzyme or the homolog from rice (Oryza sativa) exhibit resistance toward toxic fluorouridine, fluorouracil, and fluoroorotic acid, providing clear evidence for a pivotal function of URH1 as regulative in pyrimidine degradation. Moreover, mutants with increased and decreased nucleosidase activity are delayed in germination, indicating that this enzyme activity must be well balanced in the early phase of plant development. PMID- 19293371 TI - Single-walled and multi-walled carbon nanotubes promote allergic immune responses in mice. AB - The adjuvant effect of particles on allergic immune responses has been shown to increase with decreasing particle size and increasing particle surface area. Like ultrafine particles, carbon nanotubes (CNTs) have nano-sized dimensions and a large relative surface area and might thus increase allergic responses. Therefore, we examined whether single-walled (sw) and multi-walled (mw) CNTs have the capacity to promote allergic responses in mice, first in an sc injection model and thereafter in an intranasal model. Balb/cA mice were exposed to three doses of swCNT, mwCNT, as well as ultrafine carbon black particles (ufCBPs, Printex90) during sensitization with the allergen ovalbumin (OVA). Five days after an OVA booster, OVA-specific IgE, IgG1, and IgG2a antibodies in serum and the numbers of inflammatory cells and cytokine levels in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, ex vivo OVA-induced cytokine release from mediastinal lymph node (MLN) cells was measured. In separate experiments, differential cell counts were determined in BALF 24 h after a single intranasal exposure to the particles in the absence of allergen. We demonstrate that both swCNT and mwCNT together with OVA strongly increased serum levels of OVA-specific IgE, the number of eosinophils in BALF, and the secretion of Th2-associated cytokines in the MLN. On the other hand, only mwCNT and ufCBP with OVA increased IgG2a levels, neutrophil cell numbers, and tumor necrosis factor-alpha and monocyte chemoattractant protein-1 levels in BALF, as well as the acute influx of neutrophils after exposure to the particles alone. This study demonstrates that CNTs promote allergic responses in mice. PMID- 19293372 TI - Competitive binding of poly- and perfluorinated compounds to the thyroid hormone transport protein transthyretin. AB - Due to their unique surfactant properties, poly- and perfluorinated compounds (PFCs) have been extensively used and can be found all over the environment. Concern about their environmental fate and toxicological properties has initiated several research projects. In the present study, we investigated if PFCs can compete with thyroxine (T(4), i.e., the transport form of thyroid hormone) for binding to the human thyroid hormone transport protein transthyretin (TTR). Such competitive capacity may lead to decreased thyroid hormone levels as previously reported for animals exposed to PFCs. Twenty-four PFCs, together with 6 structurally similar natural fatty acids, were tested for binding capacity in a radioligand-binding assay. The binding potency decreased in the order: perfluorohexane sulfonate > perfluorooctane sulfonate/perfluorooctanoic acid > perfluoroheptanoic acid > sodium perfluoro-1-octanesulfinate > perfluorononanoic acid, with TTR binding potencies 12.5-50 times lower than the natural ligand T(4). Some lower molecular weight compounds with structural similarity to these PFCs were > 100 times less potent than T(4). Simple descriptors based on the two dimensional molecular structures of the compounds were used to visualize the chemical variation and to model the structure-activity relationship for the competitive potencies of the TTR-binding compounds. The models indicated the dependence on molecular size and functional groups but demanded a more detailed description of the chemical properties and data for validation and further quantitative structure-activity relationship (QSAR) development. Competitive binding of PFCs to TTR, as observed for human TTR in the present study, may explain altered thyroid hormone levels described for PFC-exposed rats and monkeys. Median human blood levels of the most potent TTR-binding PFCs are one to two orders of magnitude lower than concentration at 50% inhibition (IC(50)) values determined in the present study. In addition, this study contributes to the understanding of the bioaccumulation of PFCs in man and possibly in other wildlife species. PMID- 19293373 TI - Developmental chlorpyrifos and methyl parathion exposure alters radial-arm maze performance in juvenile and adult rats. AB - Although the use of organophosphate (OP) insecticides has been restricted, sufficient exposure can occur to induce detrimental neurobehavioral effects. In this study, we measured physical and reflex development and spatial learning and memory in rats repeatedly exposed to incremental doses of chlorpyrifos (CPS) and methyl parathion (MPS) from postnatal day (PND) 1 to PND21. Other than decreased body weight in the higher dosage groups, no effects on physical or reflex development were observed. Significant hippocampal cholinesterase inhibition was induced in all treatment groups for up to 19 days following exposure. Beginning on PND36, working and reference memory was tested using a 12-arm radial maze, with subject animals trained and tested 4 days a week for 4 weeks. In males, working memory was decreased with the medium and high dosage of MPS but only the high dosage of CPS; while in females, no deficits were observed. For reference memory, errors were significantly increased in males exposed to the high dosage of CPS and all dosages of MPS. In females, enhanced performance was observed within the medium and high dosages of CPS but not with MPS. These data show that repeated developmental exposure to OP insecticides can induce sex-selective alterations and long-lasting changes in spatial learning and memory formation when measured using a radial arm maze and that MPS and CPS induce different neurobehavioral outcomes. PMID- 19293374 TI - Evolution of virulence in epidemic community-associated methicillin-resistant Staphylococcus aureus. AB - Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has recently emerged worldwide. The United States, in particular, is experiencing a serious epidemic of CA-MRSA that is almost entirely caused by an extraordinarily infectious strain named USA300. However, the molecular determinants underlying the pathogenic success of CA-MRSA are mostly unknown. To gain insight into the evolution of the exceptional potential of USA300 to cause disease, we compared the phylogeny and virulence of USA300 with that of closely related MRSA clones. We discovered that the sublineage from which USA300 evolved is characterized by a phenotype of high virulence that is clearly distinct from other MRSA strains. Namely, USA300 and its progenitor, USA500, had high virulence in animal infection models and the capacity to evade innate host defense mechanisms. Furthermore, our results indicate that increased virulence in the USA300/USA500 sublineage is attributable to differential expression of core genome-encoded virulence determinants, such as phenol-soluble modulins and alpha-toxin. Notably, the fact that the virulence phenotype of USA300 was already established in its progenitor indicates that acquisition of mobile genetic elements has played a limited role in the evolution of USA300 virulence and points to a possibly different role of those elements. Thus, our results highlight the importance of differential gene expression in the evolution of USA300 virulence. This finding calls for a profound revision of our notion about CA-MRSA pathogenesis at the molecular level and has important implications for design of therapeutics directed against CA MRSA. PMID- 19293375 TI - Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine. AB - The unique monooxygenase activity of cytochrome P450cam has been attributed to coordination of a cysteine thiolate to the heme cofactor. To investigate this interaction, we replaced cysteine with the more electron-donating selenocysteine. Good yields of the selenoenzyme were obtained by bacterial expression of an engineered gene containing the requisite UGA codon for selenocysteine and a simplified yet functional selenocysteine insertion sequence (SECIS). The sulfur to-selenium substitution subtly modulates the structural, electronic, and catalytic properties of the enzyme. Catalytic activity decreases only 2-fold, whereas substrate oxidation becomes partially uncoupled from electron transfer, implying a more complex role for the axial ligand than generally assumed. PMID- 19293376 TI - Profile of Phillip Cohen. PMID- 19293377 TI - Regulation of the processivity and intracellular localization of Saccharomyces cerevisiae dynein by dynactin. AB - Dynactin, a large multisubunit complex, is required for intracellular transport by dynein; however, its cellular functions and mechanism of action are not clear. Prior studies suggested that dynactin increases dynein processivity by tethering the motor to the microtubule through its own microtubule binding domains. However, this hypothesis could not be tested without a recombinant source of dynactin. Here, we have produced recombinant dynactin and dynein in Saccharomyces cerevisiae, and examined the effect of dynactin on dynein in single-molecule motility assays. We show that dynactin increases the run length of single dynein motors, but does not alter the directionality of dynein movement. Enhancement of dynein processivity by dynactin does not require the microtubule (MT) binding domains of Nip100 (the yeast p150(Glued) homolog). Dynactin lacking these MT binding domains also supports the proper localization and function of dynein during nuclear segregation in vivo. Instead, a segment of the coiled-coil of Nip100 is required for these activities. Our results directly demonstrate that dynactin increases the processivity of dynein through a mechanism independent of microtubule tethering. PMID- 19293378 TI - The Epstein-Barr virus Bcl-2 homolog, BHRF1, blocks apoptosis by binding to a limited amount of Bim. AB - Current knowledge suggests that the balance between life and death within a cell can be controlled by the stable engagement of Bcl-2-related proapoptotic proteins such as Bak, Bax, and Bim by survival proteins such as Bcl-2. BHRF1 is a prosurvival molecule from Epstein-Barr virus that has a high degree of homology to Bcl-2. To understand how BHRF1 blocks apoptosis, BHRF1 and mutants of BHRF1 were expressed in primary cells and an IL-2-dependent T cell line. BHRF1 bound the Executioner Bak and, when cells were cultured without cytokines, BHRF1 associated with Bim. A point mutation that lost the ability to bind Bak retained its ability to bind Bim and to protect cells. This result demonstrated that it was the capacity of BHRF1 to bind Bim, not Bak, that provided protection. Interestingly, the amount of Bim bound by BHRF1 was minimal when compared with the amount of Bim induced by apoptosis. Thus, BHRF1 does not act by simply absorbing the excess Bim produced while cells prepare for death. Rather, BHRF1 may act either by binding preferentially the most lethal form of Bim or by acting catalytically on Bim to block apoptosis. PMID- 19293379 TI - New twist on nitrogen cycling in oceanic oxygen minimum zones. PMID- 19293380 TI - Interplay of alpha-synuclein binding and conformational switching probed by single-molecule fluorescence. AB - We studied the coupled binding and folding of alpha-synuclein, an intrinsically disordered protein linked with Parkinson's disease. Using single-molecule fluorescence resonance energy transfer and correlation methods, we directly probed protein membrane association, structural distributions, and dynamics. Results revealed an intricate energy landscape on which binding of alpha synuclein to amphiphilic small molecules or membrane-like partners modulates conformational transitions between a natively unfolded state and multiple alpha helical structures. Alpha-synuclein conformation is not continuously tunable, but instead partitions into 2 main classes of folding landscape structural minima. The switch between a broken and an extended helical structure can be triggered by changing the concentration of binding partners or by varying the curvature of the binding surfaces presented by micelles or bilayers composed of the lipid-mimetic SDS. Single-molecule experiments with lipid vesicles of various composition showed that a low fraction of negatively charged lipids, similar to that found in biological membranes, was sufficient to drive alpha-synuclein binding and folding, resulting here in the induction of an extended helical structure. Overall, our results imply that the 2 folded structures are preencoded by the alpha-synuclein amino acid sequence, and are tunable by small-molecule supramolecular states and differing membrane properties, suggesting novel control elements for biological and amyloid regulation of alpha-synuclein. PMID- 19293381 TI - Ethylene is an endogenous stimulator of cell division in the cambial meristem of Populus. AB - The plant hormone ethylene is an important signal in plant growth responses to environmental cues. In vegetative growth, ethylene is generally considered as a regulator of cell expansion, but a role in the control of meristem growth has also been suggested based on pharmacological experiments and ethylene overproducing mutants. In this study, we used transgenic ethylene-insensitive and ethylene-overproducing hybrid aspen (Populus tremula x tremuloides) in combination with experiments using an ethylene perception inhibitor [1 methylcyclopropene (1-MCP)] to demonstrate that endogenous ethylene produced in response to leaning stimulates cell division in the cambial meristem. This ethylene-controlled growth gives rise to the eccentricity of Populus stems that is formed in association with tension wood. PMID- 19293382 TI - Electrophysiological correlates of default-mode processing in macaque posterior cingulate cortex. AB - During the course of daily activity, our level of engagement with the world varies on a moment-to-moment basis. Although these fluctuations in vigilance have critical consequences for our thoughts and actions, almost nothing is known about the neuronal substrates governing such dynamic variations in task engagement. We investigated the hypothesis that the posterior cingulate cortex (CGp), a region linked to default-mode processing by hemodynamic and metabolic measures, controls such variations. We recorded the activity of single neurons in CGp in 2 macaque monkeys performing simple tasks in which their behavior varied from vigilant to inattentive. We found that firing rates were reliably suppressed during task performance and returned to a higher resting baseline between trials. Importantly, higher firing rates predicted errors and slow behavioral responses, and were also observed during cued rest periods when monkeys were temporarily liberated from exteroceptive vigilance. These patterns of activity were not observed in the lateral intraparietal area, an area linked to the frontoparietal attention network. Our findings provide physiological confirmation that CGp mediates exteroceptive vigilance and are consistent with the idea that CGp is part of the "default network" of brain areas associated with control of task engagement. PMID- 19293383 TI - Critical role of promoter IV-driven BDNF transcription in GABAergic transmission and synaptic plasticity in the prefrontal cortex. AB - Transcription of Bdnf is controlled by multiple promoters, which drive expression of multiple transcripts encoding for the same protein. Promoter IV contributes significantly to activity-dependent brain-derived neurotrophic factor (BDNF) transcription. We have generated promoter IV mutant mice (BDNF-KIV) by inserting a GFP-STOP cassette within the Bdnf exon IV locus. This genetic manipulation results in disruption of promoter IV-mediated Bdnf expression. BDNF-KIV animals exhibited significant deficits in GABAergic interneurons in the prefrontal cortex (PFC), particularly those expressing parvalbumin, a subtype implicated in executive function and schizophrenia. Moreover, disruption of promoter IV-driven Bdnf transcription impaired inhibitory but not excitatory synaptic transmission recorded from layer V pyramidal neurons in the PFC. The attenuation of GABAergic inputs resulted in an aberrant appearance of spike-timing-dependent synaptic potentiation (STDP) in PFC slices derived from BDNF-KIV, but not wild-type littermates. These results demonstrate the importance of promoter IV-dependent Bdnf transcription in GABAergic function and reveal an unexpected regulation of STDP in the PFC by BDNF. PMID- 19293384 TI - Activation of critical, host-induced, metabolic and stress pathways marks neutrophil entry into cystic fibrosis lungs. AB - Cystic fibrosis (CF) patients undergo progressive airway destruction caused in part by chronic neutrophilic inflammation. While opportunistic pathogens infecting CF airways can cause inflammation, we hypothesized that host-derived metabolic and stress signals would also play a role in this process. We show that neutrophils that have entered CF airways have increased phosphorylation of the eukaryotic initiation factor 4E and its partner the 4E-binding protein 1; 2 key effectors in the growth factor- and amino acid-regulated mammalian target of rapamycin (mTOR) pathway. Furthermore CF airway neutrophils display increased phosphorylation of the cAMP response element binding protein (CREB), a major transcriptional coactivator in stress signaling cascades. These active intracellular pathways are associated with increased surface expression of critical adaptor molecules, including the growth factor receptor CD114 and the receptor for advanced glycation end-products (RAGE), a CREB inducer and sensor for host-derived damage-associated molecular patterns (DAMPs). Most CF airway fluids lack any detectable soluble RAGE, an inhibitory decoy receptor for DAMPs. Concomitantly, CF airway fluids displayed high and consequently unopposed levels of S100A12; a potent mucosa- and neutrophil-derived DAMP. CF airway neutrophils also show increased surface levels of 2 critical CREB targets, the purine recycling enzyme CD39 and the multifunctional, mTOR-inducing CXCR4 receptor. This coordinated set of events occurs in all patients, even in the context of minimal airway inflammation and well-preserved lung function. Taken together, our data demonstrate an early and sustained activation of host-responsive metabolic and stress pathways upon neutrophil entry into CF airways, suggesting potential targets for therapeutic modulation. PMID- 19293385 TI - Heat-shock dependent oligomeric status alters the function of a plant-specific thioredoxin-like protein, AtTDX. AB - We found that Arabidopsis AtTDX, a heat-stable and plant-specific thioredoxin (Trx)-like protein, exhibits multiple functions, acting as a disulfide reductase, foldase chaperone, and holdase chaperone. The activity of AtTDX, which contains 3 tetratricopeptide repeat (TPR) domains and a Trx motif, depends on its oligomeric status. The disulfide reductase and foldase chaperone functions predominate when AtTDX occurs in the low molecular weight (LMW) form, whereas the holdase chaperone function predominates in the high molecular weight (HMW) complexes. Because deletion of the TPR domains results in a significant enhancement of AtTDX disulfide reductase activity and complete loss of the holdase chaperone function, our data suggest that the TPR domains of AtTDX block the active site of Trx and play a critical role in promoting the holdase chaperone function. The oligomerization status of AtTDX is reversibly regulated by heat shock, which causes a transition from LMW to HMW complexes with concomitant functional switching from a disulfide reductase and foldase chaperone to a holdase chaperone. Overexpression of AtTDX in Arabidopsis conferred enhanced heat shock resistance to plants, primarily via its holdase chaperone activity. PMID- 19293386 TI - The yin and yang of MeCP2 phosphorylation. PMID- 19293387 TI - Endocannabinoids: The silent partner of glucocorticoids in the synapse. PMID- 19293388 TI - As in humans, pregnancy increases the clearance of the protease inhibitor nelfinavir in the nonhuman primate Macaca nemestrina. AB - The apparent oral clearance of protease inhibitors (PIs) is increased in pregnant women. Although this phenomenon is reproduced in the mouse, because of the multiplicity of mouse cytochrome P450 isoforms, lack of information on their substrate and inhibitor selectivity, and lack of reagents (e.g., antibodies, purified protein), it is difficult to study the mechanistic basis of this phenomenon in this animal model. To investigate the mechanistic basis of this phenomenon in a more representative model, the nonhuman primate, we first determined whether this phenomenon could be reproduced in Macaca nemestrina, using nelfinavir as a model PI. Consistent with the human and mouse studies, we found that the apparent oral clearance of nelfinavir (NFV) in the macaques was significantly increased (3.14-fold) antepartum (n = 3) versus postpartum (n = 4). This increased apparent oral clearance was a result of an increased systemic clearance (1.9-fold) and a decreased bioavailability (approximately 45%) during pregnancy. In vitro, pregnancy significantly enhanced the rate of NFV depletion in hepatic, but not intestinal S-9 fractions. Human CYP3A inhibitors erythromycin (0.5 mM), ketoconazole (0.5 microM), and troleandomycin (0.01-1 mM), but not the CYP2C inhibitor, sulfaphenazole (3 microM), significantly inhibited the depletion of NFV in hepatic S-9 fractions and expressed rhesus CYP3A64 enzyme. Based on these data, we conclude that increased hepatic activity of NFV-metabolizing enzymes (perhaps CYP3A enzymes) results in increased clearance of PIs during pregnancy in the macaques. The M. nemestrina should be further investigated as a model to study the mechanisms by which the clearance of PIs is increased during pregnancy. PMID- 19293389 TI - Essential role of the 90-kilodalton heat shock protein in mediating nongenomic estrogen signaling in coronary artery smooth muscle. AB - Under normal physiological conditions, estrogen is a coronary vasodilator, and this response involves production of NO from endothelial cells. In addition, estrogen also stimulates NO production in coronary artery smooth muscle (CASM); however, the molecular basis for this nongenomic effect of estrogen is unclear. The purpose of this study was to investigate a potential role for the 90-kDa heat shock protein (Hsp90) in estrogen-stimulated neuronal nitric-oxide synthase (nNOS) activity in coronary artery smooth muscle. 17Beta-estradiol produced a concentration-dependent relaxation of endothelium-denuded porcine coronary arteries in vitro, and this response was attenuated by inhibiting Hsp90 function with 1 microM geldanamycin (GA) or 100 microg/ml radicicol (RAD). These inhibitors also prevented estrogen-stimulated NO production in human CASM cells and reversed the stimulatory effect of estrogen on calcium-activated potassium (BK(Ca)) channels. These functional studies indicated a role for Hsp90 in coupling estrogen receptor activation to NOS stimulation in CASM. Furthermore, coimmunoprecipitation studies demonstrated that estrogen stimulates bimolecular interaction of immunoprecipitated nNOS with Hsp90 and that either GA or RAD could inhibit this association. Blocking estrogen receptors with ICI182780 (fulvestrant) also prevented this association. These findings indicate an essential role for Hsp90 in nongenomic estrogen signaling in CASM and further suggest that Hsp90 might represent a prospective therapeutic target to enhance estrogen-stimulated cardiovascular protection. PMID- 19293390 TI - Scratching behavior and Fos expression in superficial dorsal horn elicited by protease-activated receptor agonists and other itch mediators in mice. AB - Protease-activated receptor (PAR)-2 and PAR-4 are implicated in nonhistaminergic itch. We investigated dose dependence, tachyphylaxis, and cross-tachyphylaxis of itch-associated scratching elicited by intradermal injections of PAR-2 and PAR-4 agonists, serotonin (5-hydroxytryptamine, 5-HT), and histamine in ICR mice, as well as mu-opioid modulation of PAR-2 agonist-evoked scratching. Each agent elicited dose-related increases in scratch bouts. Scratching elicited by the PAR 4 agonist and histamine both exhibited significant tachyphylaxis but no cross tachyphylaxis with each other. Scratching evoked by 5-HT did not exhibit significant tachyphylaxis but did exhibit significant cross-tachyphylaxis to scratching evoked by the PAR-2 and PAR-4 agonists and histamine. Naltrexone and high-dose morphine (10 mg/kg) attenuated PAR-2 agonist-evoked scratching, whereas lower dose morphine (1 mg/kg) had no effect. High-dose morphine also significantly increased circling behavior, which may have interfered with scratching. The PAR-2 agonist and 5-HT produced overlapping distributions of Fos like immunoreactivity in the superficial dorsal horn. These results indicate that PAR-2 and PAR-4 agonists, histamine, and 5-HT elicit itch-related scratching and activate superficial dorsal horn neurons that may participate in scratch reflex and ascending itch signaling pathways. PMID- 19293391 TI - The free radical scavenger edaravone rescues rats from cerebral infarction by attenuating the release of high-mobility group box-1 in neuronal cells. AB - Edaravone, a potent free radical scavenger, is clinically used for the treatment of cerebral infarction in Japan. Here, we examined the effects of edaravone on the dynamics of high-mobility group box-1 (HMGB1), which is a key mediator of ischemic-induced brain damage, during a 48-h postischemia/reperfusion period in rats and in oxygen-glucose-deprived (OGD) PC12 cells. HMGB1 immunoreactivity was observed in both the cytoplasm and the periphery of cells in the cerebral infarction area 2 h after reperfusion. Intravenous administration of 3 and 6 mg/kg edaravone significantly inhibited nuclear translocation and HMGB1 release in the penumbra area and caused a 26.5 +/- 10.4 and 43.8 +/- 0.5% reduction, respectively, of the total infarct area at 24 h after reperfusion. Moreover, edaravone also decreased plasma HMGB1 levels. In vitro, edaravone dose dependently (1-10 microM) suppressed OGD- and H(2)O(2)-induced HMGB1 release in PC12 cells. Furthermore, edaravone (3-30 microM) blocked HMGB1-triggered apoptosis in PC12 cells. Our findings suggest a novel neuroprotective mechanism for edaravone that abrogates the release of HMGB1. PMID- 19293392 TI - Noninvasive quantitative tomography of the therapeutic response to dexamethasone in ovalbumin-induced murine asthma. AB - Animal models of pulmonary inflammation are critical for understanding the pathophysiology of asthma and for developing new therapies. Current conventional assessments in mouse models of asthma and chronic obstructive pulmonary disease rely on invasive measures of pulmonary function and terminal characterization of cells infiltrating into the lung. The ability to noninvasively visualize and quantify the underlying biological processes in mouse pulmonary models in vivo would provide a significant advance in characterizing disease processes and the effects of therapeutics. We report the utility of near-infrared imaging agents, in combination with fluorescence molecular tomography (FMT) imaging, for the noninvasive quantitative imaging of mouse lung inflammation in an ovalbumin (OVA) induced chronic asthma model. BALB/c mice were intraperitoneally sensitized with OVA-Alum (aluminum hydroxide) at days 0 and 14, followed by daily intranasal challenge with OVA in phosphate-buffered saline from days 21 to 24. Dexamethasone and control therapies were given intraperitoneally 4 h before each intranasal inhalation of OVA from days 21 to 24. Twenty-four hours before imaging, the mice were injected intravenously with 5 nmol of the cathepsin-activatable fluorescent agent, ProSense 680. Quantification by FMT revealed in vivo cysteine protease activity within the lung associated with the inflammatory eosinophilia, which decreased in response to dexamethasone treatment. Results were correlated with in vitro laboratory tests (bronchoalveolar lavage cell analysis and immunohistochemistry) and revealed good correlation between these measures and quantification of ProSense 680 activation. We have demonstrated the ability of FMT to noninvasively visualize and quantify inflammation in the lung and monitor therapeutic efficacy in vivo. PMID- 19293393 TI - Epidermal growth factor receptor pathway substrate 8 (Eps8) is a novel regulator of cell adhesion and the blood-testis barrier integrity in the seminiferous epithelium. AB - In the seminiferous epithelium, Eps8 is localized to actin-based cell junctions at the blood-testis barrier (BTB) and the apical ectoplasmic specialization (ES) in stage V-VI tubules but is considerably diminished in stage VIII tubules. Eps8 down-regulation coincides with the time of BTB restructuring and apical ES disassembly, implicating the role of Eps8 in cell adhesion. Its involvement in Sertoli-germ cell adhesion was substantiated in studies using an in vivo animal model by treating rats with 1-(2,4-dichlorobenzy)-1H-indazole-3-carbohydrazide (adjudin) to induce anchoring junction restructuring, during which Eps8 disappeared at the apical ES before germ cell departure. In Sertoli cell cultures with established permeability barrier mimicking the BTB in vivo, the knockdown of Eps8 by RNAi led to F-actin disorganization and the mislocalization of the tight junction proteins occludin and ZO-1, suggesting the function of Eps8 in maintaining BTB integrity. In vivo knockdown of Eps8 in the testis caused germ cell sloughing and BTB damage, concomitant with occludin mislocalization, further validating that Eps8 is a novel regulator of cell adhesion and BTB integrity in the seminiferous epithelium. PMID- 19293394 TI - Phase II study of imatinib mesylate for recurrent meningiomas (North American Brain Tumor Consortium study 01-08). AB - Platelet-derived growth factor (PDGF) and its receptors (PDGFR) are frequently coexpressed in meningiomas, potentially contributing to their pathogenesis. The North American Brain Tumor Consortium conducted a phase II study to evaluate the therapeutic potential of imatinib mesylate (Gleevec), a PDGFR inhibitor, in patients with recurrent meningiomas. Patients were stratified into benign (WHO grade I) meningiomas or atypical (WHO grade II) and malignant (WHO grade III) meningiomas. The primary end point was 6-month progression-free survival (6M PFS). Patients requiring enzyme-inducing antiepileptic drugs were ineligible. Patients received imatinib at a dose of 600 mg/day for the first 4-week cycle and then gradually increased to 800 mg/day for subsequent cycles, if there were no unacceptable toxicities. Plasma concentrations of imatinib and its active metabolite, CGP74588, were assessed. Twenty-three heavily pretreated patients were enrolled into the study (13 benign, 5 atypical, and 5 malignant meningiomas), of whom 22 were eligible. The study was closed prematurely due to slow accrual. Tissue was available only from a minority of patients, but in these specimens there was uniform distribution of PDGFR, the drug target. Imatinib was generally well tolerated. Of 19 patients evaluable for response, 10 progressed at the first scan, and 9 were stable. There were no complete or partial responses. Overall median PFS was 2 months (range, 0.7-34 months); 6M-PFS was 29.4%. For benign meningiomas, median PFS was 3 months (range, 1.1-34 months); 6M-PFS was 45%. For atypical and malignant meningiomas, median PFS was 2 months (range, 0.7 3.7 months); 6M-PFS was 0%. Cycle 1 trough concentrations of imatinib and CGP74588 were 2,129 +/- 1,600 ng/ml and 517 +/- 326 ng/ml, respectively. Single agent imatinib was well tolerated but had no significant activity in recurrent meningiomas. Trough plasma concentrations of imatinib exceeded those associated with imatinib activity in chronic myelogenous leukemia. PMID- 19293395 TI - Gonadal hormones modulate the dendritic spine densities of primary cortical pyramidal neurons in adult female rat. AB - Adult dendritic arbors and spines can be modulated by environment and gonadal hormones that have been reported to affect also those of hippocampal and prefrontal cortical neurons. Here we investigated whether female gonadal hormones and estrous cycle alter the dendrites of primary cortical neurons. We employed intracellular dye injection in semifixed brain slices and 3-dimensional reconstruction to study the dendritic arbors and spines of the major cortical output cells, layer III and V pyramidal neurons, during different stages of the estrous cycle. Dendritic spines of both pyramidal neurons were more numerous during proestrus than estrus and diestrus, whereas dendritic arbors remained unaffected. Ovariohysterectomy (OHE) reduced dendritic spines by 24-30% in 2 weeks, whereas subcutaneous estrogen or progesterone supplement restored it to normal estrous/diestrous level in 14 days; neither treatment affected the dendritic arbors. Reduction of dendritic spines following OHE was associated with decrease of PSD-95 suggesting decrease of excitatory synapses. Thus, fluctuation of gonadal hormones during the female sex cycle is likely to modulate primary cortical functions and loss of gonadal hormones for instance following menopause might compromise cortical function, and the effect could be reversed by exogenous female sex hormones. PMID- 19293396 TI - Structural correlates of semantic and phonemic fluency ability in first and second languages. AB - Category and letter fluency tasks are commonly used clinically to investigate the semantic and phonological processes central to speech production, but the neural correlates of these processes are difficult to establish with functional neuroimaging because of the relatively unconstrained nature of the tasks. This study investigated whether differential performance on semantic (category) and phonemic (letter) fluency in neurologically normal participants was reflected in regional gray matter density. The participants were 59 highly proficient speakers of 2 languages. Our findings corroborate the importance of the left inferior temporal cortex in semantic relative to phonemic fluency and show this effect to be the same in a first language (L1) and second language (L2). Additionally, we show that the pre-supplementary motor area (pre-SMA) and head of caudate bilaterally are associated with phonemic more than semantic fluency, and this effect is stronger for L2 than L1 in the caudate nuclei. To further validate these structural results, we reanalyzed previously reported functional data and found that pre-SMA and left caudate activation was higher for phonemic than semantic fluency. On the basis of our findings, we also predict that lesions to the pre-SMA and caudate nuclei may have a greater impact on phonemic than semantic fluency, particularly in L2 speakers. PMID- 19293397 TI - New horizons for the subplate zone and its pioneering neurons. AB - Transitional neuronal layers are a hallmark of the prenatal and neonatal brain yet their contribution to the development of higher functions is not clear. Evidence accumulated over the last 3 decades shows that early connectivity and functional activity start in a transitional layer called the subplate zone (SPZ). The SPZ is host to a heterogeneous population of neurons and its evolutionary complexity peaked in the human brain. In this issue of Cerebral Cortex, three reports (Hoerder-Suabedissen et al., 2008; McKellar and Shatz, 2008; Moore et al., 2008) present new data and evidence in three species (mouse, rat, human) as to the function of the SPZ, to the heterogeneity of its cellular composition, and to the genetic basis of its development. PMID- 19293398 TI - Reading and subcortical auditory function. AB - Although it is largely agreed that phonological processing deficits are a major cause of poor reading, the neural origins of phonological processing are not well understood. We now show, for the first time, that phonological decoding, measured with a test of single-nonword reading, is significantly correlated with the timing of subcortical auditory processing and also, to a lesser extent, with the robustness of subcortical representation of the harmonic content of speech, but not with pitch encoding. The relationships we observe between reading and subcortical processing fall along a continuum, with poor readers at one end and good readers at the other. These data suggest that reading skill may depend on the integrity of subcortical auditory mechanisms and are consistent with the idea that subcortical representation of the acoustic features of speech may play a role in normal reading as well as in the development of reading disorders. These data establish a significant link between subcortical auditory function and reading, thereby contributing to the understanding of the biological bases of reading. At a more general level, these findings are among the first to establish a direct relationship between subcortical sensory function and a specific cognitive skill (reading). We argue that this relationship between cortical and subcortical function could be shaped during development by the corticofugal pathway and that this cortical-subcortical link could contribute to the phonological processing deficits experienced by poor readers. PMID- 19293399 TI - Odorant receptors from the light brown apple moth (Epiphyas postvittana) recognize important volatile compounds produced by plants. AB - Moths recognize a wide range of volatile compounds, which they use to locate mates, food sources, and oviposition sites. These compounds are recognized by odorant receptors (OR) located within the dendritic membrane of sensory neurons that extend into the lymph of sensilla, covering the surface of insect antennae. We have identified 3 genes encoding ORs from the tortricid moth, Epiphyas postvittana, a pest of horticulture. Like Drosophila melanogaster ORs, they contain 7 transmembrane helices with an intracellular N-terminus, an orientation in the plasma membrane opposite to that of classical GPCRs. EpOR2 is orthologous to the coreceptor Or83b from D. melanogaster. EpOR1 and EpOR3 both recognize a range of terpenoids and benzoates produced by plants. Of the compounds tested, EpOR1 shows the best sensitivity to methyl salicylate [EC(50) = 1.8 x 10(-12) M], a common constituent of floral scents and an important signaling compound produced by plants when under attack from insects and pathogens. EpOR3 best recognizes the monoterpene citral to low concentrations [EC(50) = 1.1 x 10(-13) M]. Citral produces the largest amplitude electrophysiological responses in E. postvittana antennae and elicits repellent activity against ovipositing female moths. Orthologues of EpOR3 were found across 6 families within the Lepidoptera, suggesting that the ability to recognize citral may underpin an important behavior. PMID- 19293400 TI - Different models to mobilize peer support to improve diabetes self-management and clinical outcomes: evidence, logistics, evaluation considerations and needs for future research. AB - Much of diabetes care needs to be carried out by patients between office visits with their health care providers. Yet, many patients face difficulties carrying out these tasks. In addition, many adults with diabetes cannot count on effective support from their families and friends to help them with their self-management. Peer support programmes are a promising approach to enhance social and emotional support, assist patients in daily management and living with diabetes and promote linkages to clinical care. This background paper provides a brief overview of different approaches to mobilize peer support for diabetes self-management support, discusses evidence to date on the effectiveness of each of these models, highlights logistical and evaluation issues for each model and concludes with a discussion of directions for future research in this area. PMID- 19293401 TI - Proteolytic generation of kinins in tissues infected by Trypanosoma cruzi depends on CXC chemokine secretion by macrophages activated via Toll-like 2 receptors. AB - Previous analysis of the endogenous innate signals that steer T cell-dependent immunity in mice acutely infected by the protozoan Trypanosoma cruzi revealed that bradykinin (BK) or lysyl-BK, i.e., the short-lived peptides excised from plasma-borne kininogens through the activity of cruzipain, induces dendritic cell maturation via BK B(2) receptors (B(2)R). Here, we used the s.c. model of T. cruzi infection to study the functional interplay of TLR2, CXCR2, and B(2)R in edema development. Using intravital microscopy, we found that repertaxin (CXCR2 antagonist) blocked tissue-culture trypomastigotes (TCT)-induced plasma leakage and leukocyte accumulation in the hamster cheek pouch topically exposed to TCT. Furthermore, we found that TCT-evoked paw edema in BALB/c mice was blocked by repertaxin or HOE-140 (B(2)R antagonist), suggesting that CXCR2 propels the extravascular activation of the kinin/B(2)R pathway. We then asked if TLR2 mediated sensing of TCT by innate sentinel cells could induce secretion of CXC chemokines, which would then evoke neutrophil-dependent plasma leakage via the CXCR2/B(2)R pathway. Consistent with this notion, in vitro studies revealed that TCT induce robust secretion of CXC chemokines by resident macrophages in a TLR2 dependent manner. In contrast, TLR2(+/+) macrophages stimulated with insect derived metacyclic trypomastigotes or epimastigotes, which lack the developmentally regulated TLR2 agonist displayed by TCT, failed to secrete keratinocyte-derived chemokine/MIP-2. Collectively, these results suggest that secretion of CXC chemokines by innate sentinel cells links TLR2-dependent recognition of TCT to the kinin system, a proteolytic web that potently amplifies vascular inflammation and innate immunity through the extravascular release of BK. PMID- 19293402 TI - Lnk inhibits myeloproliferative disorder-associated JAK2 mutant, JAK2V617F. AB - The JAK2 mutation JAK2V617F is found frequently in patients with myeloproliferative disorders (MPD) and transforms hematopoietic cells to cytokine independent proliferation when expressed with specific cytokine receptors. The Src homology 2 (SH2) and pleckstrin homology (PH) domain-containing adaptor protein Lnk (SH2B3) is a negative regulator of hematopoietic cytokine signaling. Here, we show that Lnk is a potent inhibitor of JAK2V617F constitutive activity. Lnk down-regulates JAK2V617F-mediated signaling and transformation in hematopoietic Ba/F3-erythropoietin receptor cells. Furthermore, in CFU assays, Lnk-deficient murine bone marrow cells are significantly more sensitive to transformation by JAK2V617F than wild-type (WT) cells. Lnk, through its SH2 and PH domains, interacts with WT and mutant JAK2 and is phosphorylated by constitutively activated JAK2V617F. Finally, we found that Lnk levels are high in CD34(+) hematopoietic progenitors from MPD patients and that Lnk expression is induced following JAK2 activation. Our data suggest that JAK2V617F is susceptible to endogenous negative-feedback regulation, providing new insights into the molecular pathogenesis of MPD. PMID- 19293403 TI - The cover. Spring. PMID- 19293404 TI - A piece of my mind. Apostrophe. PMID- 19293405 TI - Soldier suicide rates continue to rise: military, scientists work to stem the tide. PMID- 19293406 TI - Report: Growth of medical schools brings opportunity to redefine their mission. PMID- 19293407 TI - Report aims to reboot health care IT. PMID- 19293408 TI - JAMA editor honored as champion of integrity in scientific research and publishing. PMID- 19293409 TI - Surveillance of sudden death after myocardial infarction. PMID- 19293410 TI - Uninsured patients and emergency department use in the United States. PMID- 19293411 TI - Analyzing patient case mix and hospital rankings. PMID- 19293412 TI - Testing for celiac disease in patients with symptoms of irritable bowel syndrome. PMID- 19293413 TI - ACAT inhibition and progression of carotid atherosclerosis in patients with familial hypercholesterolemia: the CAPTIVATE randomized trial. AB - CONTEXT: Inhibition of acyl coenzyme A:cholesterol acyltransferase (ACAT), an intracellular enzyme involved in cholesterol accumulation, with pactimibe was developed to assist in the prevention of cardiovascular disease. OBJECTIVE: To evaluate the efficacy and safety of pactimibe in inhibition of atherosclerosis. DESIGN, SETTING, AND PATIENTS: A prospective, randomized, stratified, double blind, placebo-controlled study (Carotid Atherosclerosis Progression Trial Investigating Vascular ACAT Inhibition Treatment Effects [CAPTIVATE]) of 892 patients heterozygous for familial hypercholesterolemia conducted at 40 lipid clinics in the United States, Canada, Europe, South Africa, and Israel between February 1, 2004, and December 31, 2005. Study was terminated on October 26, 2005. INTERVENTION: Participants received either 100 mg/d of pactimibe (n = 443) or matching placebo (n = 438), in addition to standard lipid-lowering therapy. MAIN OUTCOME MEASURES: Carotid atherosclerosis, assessed by ultrasound carotid intima-media thickness (CIMT), at baseline, 12, 18, and 24 months. Maximum CIMT was the primary end point and mean CIMT the secondary end point. RESULTS: Because pactimibe failed to show efficacy in the intravascular coronary ultrasound ACTIVATE study, the CAPTIVATE study was terminated prematurely after a follow-up of 15 months. After 6 months of treatment with pactimibe, low-density lipoprotein cholesterol increased by 7.3% (SD, 23%) compared with 1.4% (SD, 28%) in the placebo group (P = .001). The carotid ultrasonographic scans of the 716 patients with at least 2 scans and obtained at least 40 weeks apart were analyzed. Maximum CIMT measurements did not show a pactimibe treatment effect (difference, 0.004 mm; 95% confidence interval [CI], -0.023 to 0.015 mm; P = .64); however, the less variable mean CIMT measurement revealed an increase of 0.014 mm (95% CI, -0.027 to 0.000 mm; P = .04) in patients administered pactimibe vs placebo. Major cardiovascular events (cardiovascular death, myocardial infarction, and stroke) occurred more often in patients administered pactimibe vs placebo (10/443 [2.3%] vs 1/438 [0.2%]; P = .01). CONCLUSIONS: In patients with familial hypercholesterolemia, pactimibe had no effect on atherosclerosis as assessed by changes in maximum CIMT compared with placebo but was associated with an increase in mean CIMT as well as increased incidence of major cardiovascular events. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00151788. PMID- 19293414 TI - Religious coping and use of intensive life-prolonging care near death in patients with advanced cancer. AB - CONTEXT: Patients frequently rely on religious faith to cope with cancer, but little is known about the associations between religious coping and the use of intensive life-prolonging care at the end of life. OBJECTIVE: To determine the way religious coping relates to the use of intensive life-prolonging end-of-life care among patients with advanced cancer. DESIGN, SETTING, AND PARTICIPANTS: A US multisite, prospective, longitudinal cohort of 345 patients with advanced cancer, who were enrolled between January 1, 2003, and August 31, 2007. The Brief RCOPE assessed positive religious coping. Baseline interviews assessed psychosocial and religious/spiritual measures, advance care planning, and end-of-life treatment preferences. Patients were followed up until death, a median of 122 days after baseline assessment. MAIN OUTCOME MEASURES: Intensive life-prolonging care, defined as receipt of mechanical ventilation or resuscitation in the last week of life. Analyses were adjusted for demographic factors significantly associated with positive religious coping and any end-of-life outcome at P < .05 (ie, age and race/ethnicity). The main outcome was further adjusted for potential psychosocial confounders (eg, other coping styles, terminal illness acknowledgment, spiritual support, preference for heroics, and advance care planning). RESULTS: A high level of positive religious coping at baseline was significantly associated with receipt of mechanical ventilation compared with patients with a low level (11.3% vs 3.6%; adjusted odds ratio [AOR], 2.81 [95% confidence interval {CI}, 1.03-7.69]; P = .04) and intensive life-prolonging care during the last week of life (13.6% vs 4.2%; AOR, 2.90 [95% CI, 1.14-7.35]; P = .03) after adjusting for age and race. In the model that further adjusted for other coping styles, terminal illness acknowledgment, support of spiritual needs, preference for heroics, and advance care planning (do-not-resuscitate order, living will, and health care proxy/durable power of attorney), positive religious coping remained a significant predictor of receiving intensive life-prolonging care near death (AOR, 2.90 [95% CI, 1.07-7.89]; P = .04). CONCLUSIONS: Positive religious coping in patients with advanced cancer is associated with receipt of intensive life-prolonging medical care near death. Further research is needed to determine the mechanisms for this association. PMID- 19293416 TI - Self-care of physicians caring for patients at the end of life: "Being connected... a key to my survival". AB - Physicians providing end-of-life care are subject to a variety of stresses that may lead to burnout and compassion fatigue at both individual and team levels. Through the story of an oncologist, we discuss the prodromal symptoms and signs leading to burnout and compassion fatigue and present the evidence for prevention. We define and discuss factors that contribute to burnout and compassion fatigue and consider factors that may mitigate burnout. We explore the practice of empathy and discuss an approach for physicians to maximize wellness through self-awareness in the setting of caring for patients with end-stage illness. Finally, we discuss some practical applications of self-care in the workplace. PMID- 19293415 TI - Effects of modafinil on dopamine and dopamine transporters in the male human brain: clinical implications. AB - CONTEXT: Modafinil, a wake-promoting drug used to treat narcolepsy, is increasingly being used as a cognitive enhancer. Although initially launched as distinct from stimulants that increase extracellular dopamine by targeting dopamine transporters, recent preclinical studies suggest otherwise. OBJECTIVE: To measure the acute effects of modafinil at doses used therapeutically (200 mg and 400 mg given orally) on extracellular dopamine and on dopamine transporters in the male human brain. DESIGN, SETTING, AND PARTICIPANTS: Positron emission tomography with [(11)C]raclopride (D(2)/D(3) radioligand sensitive to changes in endogenous dopamine) and [(11)C]cocaine (dopamine transporter radioligand) was used to measure the effects of modafinil on extracellular dopamine and on dopamine transporters in 10 healthy male participants. The study took place over an 8-month period (2007-2008) at Brookhaven National Laboratory. MAIN OUTCOME MEASURES: Primary outcomes were changes in dopamine D(2)/D(3) receptor and dopamine transporter availability (measured by changes in binding potential) after modafinil when compared with after placebo. RESULTS: Modafinil decreased mean (SD) [(11)C]raclopride binding potential in caudate (6.1% [6.5%]; 95% confidence interval [CI], 1.5% to 10.8%; P = .02), putamen (6.7% [4.9%]; 95% CI, 3.2% to 10.3%; P = .002), and nucleus accumbens (19.4% [20%]; 95% CI, 5% to 35%; P = .02), reflecting increases in extracellular dopamine. Modafinil also decreased [(11)C]cocaine binding potential in caudate (53.8% [13.8%]; 95% CI, 43.9% to 63.6%; P < .001), putamen (47.2% [11.4%]; 95% CI, 39.1% to 55.4%; P < .001), and nucleus accumbens (39.3% [10%]; 95% CI, 30% to 49%; P = .001), reflecting occupancy of dopamine transporters. CONCLUSIONS: In this pilot study, modafinil blocked dopamine transporters and increased dopamine in the human brain (including the nucleus accumbens). Because drugs that increase dopamine in the nucleus accumbens have the potential for abuse, and considering the increasing use of modafinil, these results highlight the need for heightened awareness for potential abuse of and dependence on modafinil in vulnerable populations. PMID- 19293417 TI - Impairments in endocannabinoid signaling and depressive illness. PMID- 19293418 TI - Social policy as health policy. PMID- 19293419 TI - Action on health disparities in the United States: commission on social determinants of health. PMID- 19293420 TI - Consciousness, coma, and brain death--2009. PMID- 19293421 TI - JAMA patient page. Brain death. PMID- 19293422 TI - Protection against aflatoxin B1 in rat--a new look at the link between toxicity, carcinogenicity, and metabolism. PMID- 19293423 TI - The accountability conundrum: staying focused, delivering results: a report on the UHC 2008 quality and safety fall forum. PMID- 19293424 TI - Alcohol consumption and decreased risk of non-Hodgkin lymphoma: role of mTOR dysfunction. AB - Several epidemiologic studies support the emerging paradigm that current alcohol consumers have decreased risk of most types of non-Hodgkin lymphoma. The observed lower risk among people who drank alcohol does not seem to vary with beverage type. The mechanisms accounting for alcohol-induced decrease in the incidence of lymphomas remain largely unknown. We demonstrate that low-dose chronic exposure to ethanol inhibits mammalian target of rapamycin (mTOR) C1 complex formation, resulting in decreased phosphorylation events involved in mTOR pathway signaling in a lymphoid-tissue specific manner. These changes in mTOR signaling lead to a decrease in eIF4E associated with the translation initiation complex and a repression of global cap-dependent synthesis in both lymphoma cell lines and normal donor lymphocytes. We show that chronic exposure of ethanol at physiologically relevant concentrations in a xenograft model results in a striking inhibition of lymphoma growth. Our data support a paradigm in which chronic ethanol exposure inhibits mTOR signaling in lymphocytes with a significant repression of cap-dependent translation, reducing the tumorigenic capacity of non-Hodgkin lymphoma in a human xenograft model. The ethanol-mediated repression of mTOR signaling coupled with decreased in vivo lymphoma growth underscore the critical role of mTOR signaling and translation in lymphoma. PMID- 19293425 TI - Regulation of iron homeostasis in anemia of chronic disease and iron deficiency anemia: diagnostic and therapeutic implications. AB - The anemia of chronic disease (ACD) is characterized by macrophage iron retention induced by cytokines and the master regulator hepcidin. Hepcidin controls cellular iron efflux on binding to the iron export protein ferroportin. Many patients, however, present with both ACD and iron deficiency anemia (ACD/IDA), the latter resulting from chronic blood loss. We used a rat model of ACD resulting from chronic arthritis and mimicked ACD/IDA by additional phlebotomy to define differing iron-regulatory pathways. Iron retention during inflammation occurs in macrophages and the spleen, but not in the liver. In rats and humans with ACD, serum hepcidin concentrations are elevated, which is paralleled by reduced duodenal and macrophage expression of ferroportin. Individuals with ACD/IDA have significantly lower hepcidin levels than ACD subjects, and ACD/IDA persons, in contrast to ACD subjects, were able to absorb dietary iron from the gut and to mobilize iron from macrophages. Circulating hepcidin levels affect iron traffic in ACD and ACD/IDA and are more responsive to the erythropoietic demands for iron than to inflammation. Hepcidin determination may aid to differentiate between ACD and ACD/IDA and in selecting appropriate therapy for these patients. PMID- 19293426 TI - The presence of JAK2V617F mutation in the liver endothelial cells of patients with Budd-Chiari syndrome. AB - Patients with myeloproliferative disorders are at a high risk of developing thrombotic events. Several investigators have hypothesized that endothelial cell (EC) abnormalities might contribute to this prothrombotic state. Budd-Chiari syndrome (BCS) and portal vein thrombosis have been reported to be associated with JAK2V617F-positive hematopoiesis. We explored whether JAK2V617F was present in ECs in the vessels of polycythemia vera (PV) patients with BCS using laser capture microdissection followed by nested polymerase chain reaction or reverse transcribed polymerase chain reaction. The ECs of the 2 BCS patients with PV were homozygous for the JAK2V617F and were shown to express transcripts characteristic of ECs but not hematopoietic cells. ECs of the other BCS patient with PV and 2 patients with hepatoportal sclerosis without PV contained exclusively wild-type JAK2. The presence of JAK2V617F in both ECs and hematopoietic cells belonging to BCS patients with PV indicate that ECs in PV are involved by the malignant process and that in a subpopulation of the patients the disease might originate from a common cell of origin for hematopoietic and ECs. PMID- 19293427 TI - Cortisol and epinephrine control opposing circadian rhythms in T cell subsets. AB - Pronounced circadian rhythms in numbers of circulating T cells reflect a systemic control of adaptive immunity whose mechanisms are obscure. Here, we show that circadian variations in T cell subpopulations in human blood are differentially regulated via release of cortisol and catecholamines. Within the CD4(+) and CD8(+) T cell subsets, naive cells show pronounced circadian rhythms with a daytime nadir, whereas (terminally differentiated) effector CD8(+) T cell counts peak during daytime. Naive T cells were negatively correlated with cortisol rhythms, decreased after low-dose cortisol infusion, and showed highest expression of CXCR4, which was up-regulated by cortisol. Effector CD8(+) T cells were positively correlated with epinephrine rhythms, increased after low-dose epinephrine infusion, and showed highest expression of beta-adrenergic and fractalkine receptors (CX3CR1). Daytime increases in cortisol via CXCR4 probably act to redistribute naive T cells to bone marrow, whereas daytime increases in catecholamines via beta-adrenoceptors and, possibly, a suppression of fractalkine signaling promote mobilization of effector CD8(+) T cells from the marginal pool. Thus, activation of the major stress hormones during daytime favor immediate effector defense but diminish capabilities for initiating adaptive immune responses. PMID- 19293428 TI - Bacterial FIC Proteins AMP Up Infection. AB - Proteins containing FIC (filamentation induced by cyclic adenosine monophosphate) domains are found in both prokaryotic and eukaryotic organisms, but their function has remained elusive. Recent studies indicate that bacterial FIC domain containing proteins disrupt host cell processes after being delivered into eukaryotic host cells: The Vibrio parahaemolyticus VopS protein interferes with Rho guanine triphosphatase (GTPase) function, and the Legionella pneumophila AnkX protein disrupts the microtubule-dependent transport of vesicles. Analysis of the VopS protein revealed that the FIC domain covalently modifies Rac by transferring adenosine 5'-monophosphate (AMP) to a threonine residue in the switch 1 region of the protein. Thus, FIC domain-mediated AMPylation is involved in the posttranslational regulation of protein function, and this activity has been subverted by microbial pathogens to modulate cellular functions during infection. PMID- 19293430 TI - Measurement of phosphorylated extracellular signal-regulated kinase 1 and 2 in an undergraduate teaching laboratory with ALPHAscreen technology. AB - Teaching the practical aspects of signal transduction to large undergraduate classes can be challenging when there is only a finite time frame in which to engage in laboratory activities. This teaching resource describes the use of bead based ALPHAscreen technology for a class of 300 second-year biochemistry students, exposing the next generation of researchers to cutting-edge technology. Although in this case phosphorylated extracellular signal-regulated kinase 1 and 2 were measured, this technology is applicable to the measurement of many different signaling components. This resource provides a practical guide for instructors and exemplifies how such traditionally high-throughput research technologies can be used as teaching tools. PMID- 19293429 TI - Obesity increases vascular senescence and susceptibility to ischemic injury through chronic activation of Akt and mTOR. AB - Obesity and age are important risk factors for cardiovascular disease. However, the signaling mechanism linking obesity with age-related vascular senescence is unknown. Here we show that mice fed a high-fat diet show increased vascular senescence and vascular dysfunction compared to mice fed standard chow and are more prone to peripheral and cerebral ischemia. All of these changes involve long term activation of the protein kinase Akt. In contrast, mice with diet-induced obesity that lack Akt1 are resistant to vascular senescence. Rapamycin treatment of diet-induced obese mice or of transgenic mice with long-term activation of endothelial Akt inhibits activation of mammalian target of rapamycin (mTOR) rictor complex 2 and Akt, prevents vascular senescence without altering body weight, and reduces the severity of limb necrosis and ischemic stroke. These findings indicate that long-term activation of Akt-mTOR signaling links diet induced obesity with vascular senescence and cardiovascular disease. PMID- 19293431 TI - Correlates of undefined cause of injury coded mortality data in Australia. AB - The objective of this research was to identify the level of detail regarding the external causes of death in Australia and ascertain problematic areas where data quality improvement efforts may be focused. The 2003 national mortality dataset of 12,591 deaths with an external cause of injury as the underlying cause of death (UCOD) or multiple cause of death (MCOD) based on ICD-10 code assignment from death certificate information was obtained. Logistic regression models were used to examine the precision of coded external cause of injury data. It was found that overall, accidents were the most poorly defined of all intent code blocks with over 30% of accidents being undefined, representing 2,314 deaths in 2003. More undefined codes were identified in MCOD data than for UCOD data. Deaths certified by doctors were more likely to use undefined codes than deaths certified by a coroner or government medical office. To improve the quality of external cause of injuries leading to or associated with death, certifiers need to be made aware of the importance of documenting all information pertaining to the cause of the injury and the intent behind the incident, either through education or more explicit instructions on the death certificate and accompanying instructional materials. It is important that researchers are aware of the validity of the data when they make interpretations as to the underlying causes of fatal injuries and causes of injury associated with deaths. PMID- 19293432 TI - The quality of injury data from hospital records in Vietnam. AB - The objective of this research was to examine the level of agreement of coders for ICD-10 coding of injury discharges in Danang General Hospital in Vietnam. Two hundred and five medical records of children hospitalised in this facility due to injury were randomly selected and recoded. Information from medical records abstracted by two trained staff was recoded by external coders in Hanoi and in Australia, using ICD-10. The completeness and detail of external cause of injury recorded in medical records was poor. Agreement between coders for injury coding was average, with 32% to 40% discrepancy in the main diagnosis codes at three character level, and 57% to 60% discrepancy at four character level, depending on which coders were being compared. It was concluded that as hospital data represent a cost-effective source of information regarding injuries, with significant costs incurred in collecting such information through special studies and censuses (especially for a developing country such as Vietnam), it is important to establish the quality and value of hospital data for injury surveillance and prevention research and to explore ways in which these data can be improved. PMID- 19293433 TI - Do we have enough information? How ICD-10-AM Activity codes measure up. AB - This research explored the usage of activity codes introduced into the International Statistical Classification of Diseases and Related Health Problems, Tenth revision, Australian Modification (ICD-10-AM) Third Edition and examined the data quality of activity coding, explicitly, completeness and specificity. Injury separations for years 2001/02 to 2005/06 specifying a 'true injury' were extracted for descriptive analyses. Part A investigated the usage of activity codes and compared the usage of the 236 activity codes available in the Activity block (U50-U73) present in the ICD-10-AM Third Edition against the 16 codes present in the second edition. Part B examined the level of completeness of external cause coding and the degree of activity coding specificity in the 2005/06 dataset. It was found that the additional activity codes were used extensively with only 46 codes seldom assigned. Codes present in the second edition were extensively used in the third and fourth editions and the new additional activity codes represent 10% of all activity codes assigned per year. All five datasets demonstrated high levels of completeness, recording completeness levels greater than 97%, where missing activity codes attributed to the majority of missing codes. Fourteen out of the 24 activity categories demonstrated a strong reliance on non-specific codes and Team ball sports and Wheeled non-motor sports illustrated that activity codes assigned lacked detail in the code. Clinicians and coders need to acknowledge the importance of quality clinical documentation for research and policy-making purposes so that circumstances surrounding injury events can be coded to the highest level of specificity to improve injury prevention and control activities. Missing activity codes and the abundance of non-specific coding hinders the usefulness of the data. PMID- 19293434 TI - The risk and consequences of clinical miscoding due to inadequate medical documentation: a case study of the impact on health services funding. AB - As coded clinical data are used in a variety of areas (e.g. health services funding, epidemiology, health sciences research), coding errors have the potential to produce far-reaching consequences. In this study the causes and consequences of miscoding were reviewed. In particular, the impact of miscoding due to inadequate medical documentation on hospital funding was examined. Appropriate reimbursement of hospital revenue in the casemix-based (output-based) funding system in the state of Victoria, Australia relies upon accurate, comprehensive, and timely clinical coding. In order to assess the reliability of these data in a Melbourne tertiary hospital, this study aimed to: (a) measure discrepancies in clinical code assignment; (b) identify resultant Diagnosis Related Group (DRG) changes; (c) identify revenue shifts associated with the DRG changes; (d) identify the underlying causes of coding error and DRG change; and (e) recommend strategies to address the aforementioned. An internal audit was conducted on 752 surgical inpatient discharges from the hospital within a six month period. In a blind audit, each episode was re-coded. Comparisons were made between the original codes and the auditor-assigned codes, and coding errors were grouped and statistically analysed by categories. Changes in DRGs and weighted inlier-equivalent separations (WIES) were compared and analysed, and underlying factors were identified. Approximately 16% of the 752 cases audited reflected a DRG change, equating to a significant revenue increase of nearly AU$575,300. Fifty-six percent of DRG change cases were due to documentation issues. Incorrect selection or coding of the principal diagnosis accounted for a further 13% of the DRG changes, and missing additional diagnosis codes for 29%. The most significant of the factors underlying coding error and DRG change was poor quality of documentation. It was concluded that the auditing process plays a critical role in the identification of causes of coding inaccuracy and, thence, in the improvement of coding accuracy in routine disease and procedure classification and in securing proper financial reimbursement. PMID- 19293435 TI - Commission on Social Determinants in Health: A piece meal move? PMID- 19293436 TI - The need for longitudinal measurements at altitude. PMID- 19293437 TI - Placental pathology in thrombophilia: establishing clinico-pathological correlations. PMID- 19293438 TI - Role of reactive oxygen species in the pathogenesis of mitochondrial DNA (mtDNA) mutations in male infertility. AB - Infertility affects about 15 per cent married couples half of which may be attributed to men with low sperm motility (asthenozoospermia), low sperm count (oligozoospermia) or abnormal sperm morphology (teratozoospermia). As mitochondria are the energy source for initiation, differentiation and function of the germ cells, mutation in mitochondrial genome can impair the formation of mature spermatozoa. Mutations in mitochondrial genome are identified in patients with fertility problems. However, mitochondria are also both the source and target of reactive oxygen species (ROS). ROS are normally generated at low levels by human spermatozoa in order to perform its physiological function. However, if the generation of these reactive free radicals overwhelm the antioxidant defense system, this can lead to oxidative stress, which is characterized by mitochondrial and nuclear genome damage. So both ROS and mtDNA mutations are considered to be the major aetiological factors in a variety of human diseases including male infertility. Identification of novel mutations in mtDNA of infertile patients with supraphysiological levels of ROS are considered to be important to gain better understanding of the aetiology of idiopathic infertility. Early detection and prompt antioxidant therapy can prevent ROS induced DNA damage. This has far reaching impact if such men opt for assisted reproductive technology (ART)/in vitro fertilization. PMID- 19293439 TI - Body composition & cardiovascular functions in healthy males acclimatized to desert & high altitude. AB - BACKGROUND & OBJECTIVES: Several physiological changes affecting physical fitness occur in humans whenever they are exposed to extremes of environments such as heat, cold and high altitude (HA). The present study was undertaken to evaluate effect of stay in desert and HA on physical fitness and body composition of physically active individuals. METHODS: Study was conducted on three groups of male soldiers (n=30 in each group) at different climatic conditions i.e., temperate (plains of north India), hot desert (Rajasthan), and HA (3600 m) in Western Himalayas. Subjects were acclimatized to hot and HA environments and had similar BMI (body mass index). Body fat, lean body mass, haemoglobin levels were determined along with, blood pressure and physical fitness index (PFI). RESULTS: The body fat of subjects at temperate, desert and HA was found to be 15.4, 12.8 and 16.9 per cent respectively. The resting heart rate and blood pressure were higher in altitude group in comparison to others. PFI score of volunteers at temperate, desert and HA were found to be 97.4 +/- 10.3, 92.4+/- 14.4 and 83.8 +/ 6.2 respectively. INTERPRETATION & CONCLUSION: A combination of different factors i.e., higher resting pulse rate, increased blood pressure and body fat may be responsible for lower PFI at HA. The observed differences in body fat content of different groups could be an adaptive feature to the environment. PMID- 19293440 TI - Placental histomorphology in unexplained foetal loss with thrombophilia. AB - BACKGROUND & OBJECTIVES: Acquired and genetic thrombotic conditions, both organ and non organ specific, are associated with increased foetal wastage. This study was carried out to examine the placenta from women with abnormal pregnancies and a history of unexplained foetal loss, and to associate with maternal thrombophilia status. METHODS: Placentas from eight women with history of unexplained foetal loss were analyzed for histopathological characteristics. All the women were simultaneously screened for the common acquired and genetic thrombophilia markers i.e., lupus anticoagulants ( LA), IgG / IgM antibodies for anticardiolipin (ACA), beta2 glycoprotein 1 (beta2GPI) and annexin V, protein C (PC), protein S (PS), antithrombin III (AT III), factor V Leiden ( FVL) mutation, prothrombin (PT) gene G20210A, methylene tetrahydrofolate reductase (MTHFR) C 677T, endothelial protein C receptor (EPCR) 23 bp insertion and plasminogen activator inhibitor ( PAI-1 4G/5G) polymorphisms RESULTS: Six of eight women were positive for one or more thrombophilia markers. The placenta in all the cases except one, showed the characteristic features of infarct fibrin deposition and calcification. Among two women who were negative for thrombophilia, one showed clear evidence of thrombus in the placental sections while the other did not show any characteristic infarcts in the placental sections. INTERPRETATION & CONCLUSION: Our findings showed that the histopathological examination of the placentas confirmed thrombophilia as the aetiological cause of thrombosis in 6 of the 8 women. The presence of thrombus in a negative thrombophilia woman suggests yet unidentified thrombophilia markers or probably non-haemostatic factors causing thrombosis. PMID- 19293441 TI - Influence of squalene feeding on plasma leptin, testosterone & blood pressure in rats. AB - BACKGROUND & OBJECTIVES: Obesity, hyperlipidaemia and hypercholesterolaemia are known risk factors in the pathogenesis of hypertension. Squalene has been shown to reduce serum cholesterol and triglycerides in dogs although its therapeutic use in high BP and obese patients has not been established. This study evaluates the effect of feeding high doses of squalene on plasma leptin, glucose, testosterone, blood pressure (BP) and body fat in rats. METHODS: Wistar rats (male, 22 days old) were randomly divided into two groups receiving either regular control diet or a squalene-containing diet. After feeding squalene for 4 wk, 10 rats each from the squalene and control groups were sacrificed and blood samples were collected for measurement of leptin, cholesterol, triglycerides and testosterone. Blood pressure was monitored weekly. RESULTS: Following squalene feeding, BP and body weight gain were lower in the squalene group. BP was significantly lower from 47 days of age in squalene fed group compared to controls. The levels of plasma leptin, glucose, cholesterol and triglycerides were significantly lower in squalene fed rats than those from the control group at 51 and 75 days of age. However, testicular weights (only 75 days) and testosterone levels were significantly higher in rats from the squalene group than those from the control group at days 51 and 75. INTERPRETATION & CONCLUSION: Our results suggested that squalene may counteract the increase in body fat, BP and levels of plasma leptin, glucose, cholesterol and triglycerides. These effects of squalene may be further explored as a likely new approach for clinical management of high BP and obesity. PMID- 19293442 TI - Effects of hypoxia inducible factor-1alpha (HIF-1alpha) on the growth & adhesion in tongue squamous cell carcinoma cells. AB - BACKGROUND & OBJECTIVES: Hypoxia-inducible factor-1 alpha (HIF-1alpha) is a central transcriptional regulator of hypoxic response. Suppression of HIF-1alpha is important for exploring hypoxia-induced pathophysiological events. This study was carried out to analyze the hypoxia-induced changes of biological characteristics in the human tongue squamous cell carcinoma cell line Tca8113 and evaluate the effects of HIF-1alpha on the phenotype of the tongue squamous cell carcinoma. METHODS: HIF-1alpha gene was silenced with synthesized short interfering ribonucleic acids (siRNA). HIF-1alpha expression was measured on mRNA level by real-time reverse transcription (RT)-PCR and protein level by Western blot and immunofluorescence. The cell cycle and apoptosis of Tca8113 cells were analyzed by FACS. The proliferation and adhesion of Tca8113 cells were determined by MTT colorimetric assay. RESULTS: Tca8113 could survive and showed a more aggressive phenotype under hypoxic condition. Exposure to hypoxia induced a prolonged elevation of HIF-1alpha protein and transfection of siRNA targeting HIF 1alpha (siRNA(HIF-1alpha)) reduced HIF-1alpha synthesis as measured on mRNA and protein level. Under normoxic or hypoxic conditions, treatment of Tca8113 cells with siRNA(HIF-1alpha) induced cell apoptosis and inhibited the growth and adhesion. INTERPRETATION & CONCLUSION: siRNA(HIF-1alpha) could attenuate the tolerance against hypoxia in Tca8113 cells and inhibit their aggressive potential. Interfering with HIF-1alpha pathways by siRNA strategy may provide a therapeutic target for human tongue squamous cell carcinomas. PMID- 19293443 TI - Asymmetrical release of interleukin-6 by cultured cerebral cortical astrocytes treated with lipopolysaccharide. AB - BACKGROUND & OBJECTIVES: The distribution of brain interleukin-6 (IL-6) may be asymmetrical both in cortex and hippocampus. While the brain asymmetry has been extensively investigated, the cellular origin of asymmetrical cytokine induction in the cortex has not been addressed. It was hypothesized that the immune function of glia cell to the inflammatory insults is asymmetrically distributed in the two brain hemispheres. To test this hypothesis, we examined the IL-6 secreting ability of the astrocytes in both the left and right neocortex treated with lipopolysaccharide(LPS) cultured in vitro. METHODS: Two groups of astrocytes cultured in vitro from the two cerebral cortices of the neonatal BALB/ c mice were selected and experimental group was treated with LPS (10 mug/ml) for 24 h. IL-6 levels were measured in both LPS-treated and untreated astrocytes. To confirm the gene array data on the secretion of IL-6 by cortical astrocytes in the left and right hemispheres, semi-quantitative reverse transcriptionpolymerase chain reaction (RT-PCR) was conducted. RESULTS: A statistically significant difference between the levels of IL-6 in cortical astrocytes in the left and right hemispheres of culture supernatants was observed (P<0.05). Cortical astrocytes in the left hemisphere had significantly increased IL-6 mRNA levels compared with cortical astrocytes in right hemisphere (P<0.05). INTERPRETATION & CONCLUSION: The results showed asymmetrical release of brain IL-6 by cerebral cortical astrocytes to the inflammatory insults both in protein and mRNA levels. PMID- 19293444 TI - Organic anion transporter protein (OATP1B1) encoded by SLCO1B1 gene polymorphism (388A>G) & susceptibility in gallstone disease. AB - BACKGROUND & OBJECTIVES: Organic anion transport protein 1B1 (OATP1B1) is a major transporter protein for bile salt uptake in the enterohepatic circulation of bile salts. As the role of SLCO1B1 gene (encodes OATP1B1 or liver specific transporter 1) 388 A>G polymorphism in susceptibility towards gallstone disease is unclear the prevalence of this polymorphism in healthy north Indian population was investigated. METHODS: Peripheral venous blood of 270 unrelated northern Indian patients with symptomatic gallstone disease and 270 unrelated healthy control subjects was screened for SLCO1B1 gene 388 A>G polymorphism by PCR-RFLP method and genotyping was done on 12 per cent polyacrylamide gel. The cross-sectional data on accrual of cases and controls were collected and odds ratio with 95 per cent CI calculated as for case-control design. RESULTS: Allele frequencies of 388 G were 45 per cent in gallstone cases and 44 per cent in controls with no statistical significance. Genotype frequencies in gallstone cases and controls for, genotype AA were 30 and 32 per cent; AG: 51 and 47 per cent and GG: 16 and 21 per cent respectively. No significant association of any allele or genotype with gallstone disease was found. INTERPRETATION & CONCLUSION: Although the prevalence of SLCO1B1 gene 388A>G polymorphism in north Indian population in high, yet this polymorphism does not appear to play a significant role in susceptibility to gallstone formation. PMID- 19293445 TI - DNA sequencing by Microseq kit targeting 16S rRNA gene for species level identification of mycobacteria. AB - BACKGROUND & OBJECTIVES: Identification of mycobacteria to the species level is of therapeutic significance. Conventional methods are laborious and time consuming so we did 16S rRNA sequencing using a commercial MicroSeq sequencing kit, which includes DNA sequencing with software package for identification and phylogenetic analysis of clinical mycobacterial isolates. METHODS: A total of 47 mycobacteria were tested by both conventional and genotypic method using commercially available MicroSeq 500 amplification kit assay. The identification was determined by comparing the 500 bp amplified product of 16S rDNA sequence to the MicroSeq database. RESULTS: The phenotypic identification was concordant with genotypic identification in 33 (70.2%) isolates of 14 Mycobacterium tuberculosis, 11 M. fortuitum, 7 M. abscessus and 1 M. duvalii. For the discrepant isolates, identification was possible only by DNA sequencing in 14 (29.7%) isolates. The 14 discrepant isolates were 5 M. farcinogenes, 3 M. genavense, 2 M. species. nov and 1 each of M. fortuitum, M. immuogenum, M. simiae and M. wolinskyi. Of these, five were uncommon species that were difficult to identify by phenotypic method. INTERPRETATION & CONCLUSION: The MicroSeq DNA sequencing is an excellent tool for species identification of mycobacteria, which reduces the turn around time, makes repeat analysis easy as compared to phenotypic identification specially for mycobacterial isolates with ambiguous biochemical profiles. PMID- 19293446 TI - Application of 16S rDNA based seminested PCR for diagnosis of acute bacterial meningitis. AB - BACKGROUND & OBJECTIVES: The diagnosis of bacterial meningitis remains a challenge to the clinician because of its rapid lethal course lacking the consistency to particular clinical signs and symptoms. Moreover, in many clinical settings use of rampant and short course antibiotic therapy prior to lumbar puncture reduces the chance of isolation of bacteria in CSF culture making the diagnosis difficult. The present study was done to evaluate a multiplex seminested PCR based method for rapid diagnosis of bacterial meningitis even after initiation of antibiotics. METHODS: A 16S rDNA based PCR technique was evaluated using universal bacterial primers to detect any bacterial pathogen in CSF samples. The simultaneous use of three species-specific primers in a multiplex and seminested PCR format was done to identify Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis within 4 h. RESULTS: Analysis of 267 CSF samples obtained from suspected cases of acute bacterial meningitis revealed 94 per cent concordance in results for conventional (Gram stain and culture) and molecular methods. Conventional techniques failed to detect five PCR positive samples where clinical diagnosis, cell count and biochemical findings of CSF supported the evidence of infection. The overall sensitivity, specificity, positive predictive value and negative predictive value of 16S rDNA PCR were 79.24, 97.6, 89.36 and 94.88 per cent respectively when culture was considered as gold standard. The detection limit of 16S rDNA PCR was determined to be 1000 cfu/ml of E. coli and 4000 cfu/ml of S. pneumoniae. INTERPRETATION & CONCLUSION: The results suggest that 16S rDNA PCR can be used as a valuable supplementary test in routine clinical practice for diagnosis of acute bacterial meningitis in hospital setting. PMID- 19293447 TI - Biochemical analysis of protein stability in human brain collected at different post-mortem intervals. AB - BACKGROUND & OBJECTIVES: Rising prevalence of neurodegenerative disorders with a steady increase in aged-population necessitates studies of the human brain to understand their pathophysiology. As animal models are not available, medical centers have established "brain banks" to provide autopsy brain samples for such research. Frozen tissues must be of optimal quality to permit molecular and protein studies. Post-mortem interval (PMI) is an important factor affecting tissue quality although its effects on brain physiology are unclear. We undertook this study to analyze the biochemical effects of PMI on protein stability in human brains collected at autopsy and stored at the brain bank of a tertiary care neurosciences institute in south India. METHODS: Different neuroanatomical areas including frontal cortex (FC), cerebellum (CB), caudate nucleus (CD) and substantia nigra (SN) from autopsy human brains (n=9) with varying PMI (4-18 h) were analyzed for pH, protein insolubility, protein oxidation/ nitration and protein expression of glial fibrillary acidic protein (GFAP), synatophysin and neurofilament (NF). Histological changes at different PMI were also assessed. RESULTS: An increase in tissue pH was noted with increasing PMI. Although there was no significant alteration in solubility of proteins, SN showed increased protein oxidation/nitration events, GFAP and NF expression with increasing PMI. No major abnormalities in cell morphology or tissue integrity were noted. Immunohistochemistry with GFAP and NF did not show any significant increase in signal in FC at high PMI. INTERPRETATION & CONCLUSION: In post-mortem human brains, although there were no gross structural changes at the tissue level with increasing PMI, biochemical events such as oxidative and nitrosative damage of cellular proteins, tissue pH could be considered as markers of tissue quality for biochemical research. Further, SN was found to be most susceptible to PMI related changes. PMID- 19293448 TI - Equitable healthcare & access to affordable medicines in India. PMID- 19293449 TI - Reviving frogs in India's freshwater environment to control mosquito-borne diseases. PMID- 19293450 TI - Malaria - change in therapeutic policy. PMID- 19293451 TI - WHO study suggests low incidence of Hib in india is due to natural immunity. PMID- 19293453 TI - Antipsychotic drugs & their side effects. PMID- 19293455 TI - Gene therapy in large animal models of human genetic diseases. Introduction. PMID- 19293456 TI - Gene therapy for lysosomal storage diseases (LSDs) in large animal models. AB - Lysosomal storage diseases (LSDs) are inherited metabolic disorders caused by deficient activity of a single lysosomal enzyme or other defects resulting in deficient catabolism of large substrates in lysosomes. There are more than 40 forms of inherited LSDs known to occur in humans, with an aggregate incidence estimated at 1 in 7,000 live births. Clinical signs result from the inability of lysosomes to degrade large substrates; because most lysosomal enzymes are ubiquitously expressed, a deficiency in a single enzyme can affect multiple organ systems. Thus LSDs are associated with high morbidity and mortality and represent a significant burden on patients, their families, the health care system, and society. Because lysosomal enzymes are trafficked by a mannose 6-phosphate receptor mechanism, normal enzyme provided to deficient cells can be localized to the lysosome to reduce and prevent storage. However, many LSDs remain untreatable, and gene therapy holds the promise for effective therapy. Other therapies for some LSDs do exist, or are under evaluation, including heterologous bone marrow or cord blood transplantation (BMT), enzyme replacement therapy (ERT), and substrate reduction therapy (SRT), but these treatments are associated with significant concerns, including high morbidity and mortality (BMT), limited positive outcomes (BMT), incomplete response to therapy (BMT, ERT, and SRT), life long therapy (ERT, SRT), and cost (BMT, ERT, SRT). Gene therapy represents a potential alternative, albeit with its own attendant concerns, including levels and persistence of expression and insertional mutagenesis resulting in neoplasia. Naturally occurring animal homologues of LSDs have been described in all common domestic animals (and in some that are less common) and these animal models play a critical role in evaluating the efficacy and safety of therapy. PMID- 19293457 TI - Gene therapy for inhereted metabolic disorders in companion animals. AB - Scientists first described inborn errors of metabolism, also termed inherited disorders of metabolism, early in the 20th century and since then have determined the biochemical and genetic bases of a great number of these disorders both in humans and in an increasing number of companion animals. The availability of metabolic screening tests has advanced the biochemical and genetic characterization in affected breeds of companion animals of inherited metabolic disorders involving amino acid, carbohydrate, fatty acid, and metal metabolism. Advances in gene therapy have led to the development of new treatments for inherited disorders of metabolism, and animal models have played a critical role in this research. For example, glycogen storage disease type Ia in dogs was highly responsive to adeno-associated viral vectormediated gene therapy, which prolonged survival and for more than a year prevented hypoglycemia during fasting. Gene therapy for other glycogen storage diseases and metabolic disorders will also be feasible. The establishment of a breeding colony and the ability to sustain affected animals are critical steps toward evaluating the safety and efficacy of gene therapy with clinically relevant endpoints. The further development of gene therapy for inherited disorders of metabolism could lead to curative therapy for affected humans and animals alike. PMID- 19293458 TI - Large animal models of neurological disorders for gene therapy. AB - he development of therapeutic interventions for genetic disorders and diseases that affect the central nervous system (CNS) has proven challenging. There has been significant progress in the development of gene therapy strategies in murine models of human disease, but gene therapy outcomes in these models do not always translate to the human setting. Therefore, large animal models are crucial to the development of diagnostics, treatments, and eventual cures for debilitating neurological disorders. This review focuses on the description of large animal models of neurological diseases such as lysosomal storage diseases, Parkinsons disease, Huntingtons disease, and neuroAIDS. The review also describes the contributions of these models to progress in gene therapy research. PMID- 19293459 TI - Protein replacement therapy and gene transfer in canine models of hemophilia A, hemophilia B, von willebrand disease, and factor VII deficiency. AB - Dogs with hemophilia A, hemophilia B, von Willebrand disease (VWD), and factor VII deficiency faithfully recapitulate the severe bleeding phenotype that occurs in humans with these disorders. The first rational approach to diagnosing these bleeding disorders became possible with the development of reliable assays in the 1940s through research that used these dogs. For the next 60 years, treatment consisted of replacement of the associated missing or dysfunctional protein, first with plasma-derived products and subsequently with recombinant products. Research has consistently shown that replacement products that are safe and efficacious in these dogs prove to be safe and efficacious in humans. But these highly effective products require repeated administration and are limited in supply and expensive; in addition, plasma-derived products have transmitted bloodborne pathogens. Recombinant proteins have all but eliminated inadvertent transmission of bloodborne pathogens, but the other limitations persist. Thus, gene therapy is an attractive alternative strategy in these monogenic disorders and has been actively pursued since the early 1990s. To date, several modalities of gene transfer in canine hemophilia have proven to be safe, produced easily detectable levels of transgene products in plasma that have persisted for years in association with reduced bleeding, and correctly predicted the vector dose required in a human hemophilia B liver-based trial. Very recently, however, researchers have identified an immune response to adeno-associated viral gene transfer vector capsid proteins in a human liver-based trial that was not present in preclinical testing in rodents, dogs, or nonhuman primates. This article provides a review of the strengths and limitations of canine hemophilia, VWD, and factor VII deficiency models and of their historical and current role in the development of improved therapy for humans with these inherited bleeding disorders. PMID- 19293460 TI - Potential large animal models for gene therapy of human genetic diseases of immune and blood cell systems. AB - Genetic mutations involving the cellular components of the hematopoietic system- red blood cells, white blood cells, and platelets--manifest clinically as anemia, infection, and bleeding. Although gene targeting has recapitulated many of these diseases in mice, these murine homologues are limited as translational models by their small size and brief life span as well as the fact that mutations induced by gene targeting do not always faithfully reflect the clinical manifestations of such mutations in humans. Many of these limitations can be overcome by identifying large animals with genetic diseases of the hematopoietic system corresponding to their human disease counterparts. In this article, we describe human diseases of the cellular components of the hematopoietic system that have counterparts in large animal species, in most cases carrying mutations in the same gene (CD18 in leukocyte adhesion deficiency) or genes in interacting proteins (DNA cross-link repair 1C protein and protein kinase, DNA-activated catalytic polypeptide in radiation-sensitive severe combined immunodeficiency). Furthermore, we describe the potential of these animal models to serve as disease specific preclinical models for testing the efficacy and safety of clinical interventions such as hematopoietic stem cell transplantation or gene therapy before their use in humans with the corresponding disease. PMID- 19293462 TI - Gene therapy in large animal models of human cardiovascular genetic disease. AB - Several naturally occurring animal models for human genetic heart diseases offer an excellent opportunity to evaluate potential novel therapies, including gene therapy. Some of these diseases--especially those that result in a structural defect during development (e.g., patent ductus arteriosus, pulmonic stenosis)- would likely be difficult to treat with a therapeutic gene transfer approach. However, the ability to transduce a significant proportion of the myocardial cells should make the various forms of inherited cardiomyopathy amenable to a therapeutic gene transfer approach. Adeno-associated virus may be the ideal vector for cardiac gene therapy since its low immunogenicity allows for stable transgene expression, a crucial factor when considering treatment of a chronic disease. Cardiomyopathies are a major cause of morbidity and mortality in both children and adults, and large animal models are available for the major forms of inherited cardiomyopathy (dilated cardiomyopathy, hypertrophic cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy). One of these animal models, juvenile dilated cardiomyopathy of Portuguese water dogs, offers an effective means to assess the efficacy of therapeutic gene transfer to alter the course of cardiomyopathy and heart failure. Correction of the abnormal metabolic processes that occur with heart failure (e.g., calcium metabolism, apoptosis) could normalize diseased myocardial function. Gene therapy may offer a promising new approach for the treatment of cardiac disease in both veterinary and human clinical settings. PMID- 19293461 TI - Gene therapy in large animal models of muscular dystrophy. AB - The muscular dystrophies are a group of genetically and phenotypically heterogeneously inherited diseases characterized by progressive muscle wasting, which can lead to premature death in severe forms such as Duchenne muscular dystrophy (DMD). In many cases they are caused by the absence of proteins that are critical components of the dystrophin-glycoprotein complex, which links the cytoskeleton and the basal lamina. There is no effective treatment for these disorders at present, but several novel strategies for replacing or repairing the defective gene are in development, with early encouraging results from animal models. We review these strategies, which include the use of stem cells of different tissue origins, gene replacement therapies mediated by various viral vectors, and transcript repair treatments using exon skipping strategies. We comment on their advantages and on limitations that must be overcome before successful application to human patients. Our focus is on studies in a clinically relevant large canine model of DMD. Recent advances in the field suggest that effective therapies for muscular dystrophies are on the horizon. Because of the complex nature of these diseases, it may be necessary to combine multiple approaches to achieve a successful treatment. PMID- 19293463 TI - AAV-mediated gene therapy for retinal disorders in large animal models. AB - Retinal gene therapy holds great promise for the treatment of inherited and noninherited blinding diseases such as retinitis pigmentosa and age-related macular degeneration. The most widely used vectors for ocular gene delivery are based on adeno-associated virus (AAV) because they elicit minimal immune responses and mediated long-term transgene expression in a variety of retinal cell types. Extensive preclinical evaluation of new strategies in large animal models is key to the development of successful gene-based therapies for the retina. Because of differences in the retinal structures among species and unique structures such as the macula and fovea in the primate retina, nonhuman primates are widely used as preclinical animal models. But the observation of inherited retinal degenerations in dogs, which share a number of clinical and pathologic similarities with humans, has led to the characterization of several canine models for retinal diseases, one of which has already responded successfully to AAV-mediated gene therapy. This article presents a review and detailed discussion of the various large animal models available for the study of AAV-mediated gene based therapies in the retina. PMID- 19293464 TI - Large animal models of genetic disease: pertinent IACUC issues. PMID- 19293465 TI - Intra-voxel heterogeneity influences the dose prescription for dose-painting with radiotherapy: a modelling study. AB - The purpose of this study was to increase the potential of dose redistribution by incorporating estimates of oxygen heterogeneity within imaging voxels for optimal dose determination. Cellular oxygen tension (pO(2)) distributions were estimated for imaging-size-based voxels by solving oxygen diffusion-consumption equations around capillaries placed at random locations. The linear-quadratic model was used to determine cell survival in the voxels as a function of pO(2) and dose. The dose distribution across the tumour was optimized to yield minimal survival after 30 x 2 Gy fractions by redistributing the dose based on differences in oxygen levels. Eppendorf data of a series of 69 tumours were used as a surrogate of what might be expected from oxygen imaging datasets. Dose optimizations were performed both taking into account cellular heterogeneity in oxygenation within voxels and assuming a homogeneous cellular distribution of oxygen. Our simulations show that dose redistribution based on derived cellular oxygen distributions within voxels result in dose distributions that require less total dose to obtain the same degree of cell kill as dose distributions that were optimized with a model that considered voxels as homogeneous with respect to oxygen. Moderately hypoxic tumours are expected to gain most from dose redistribution. Incorporating cellular-based distributions of radiosensitivity into dose-planning algorithms theoretically improves the potential gains from dose redistribution algorithms. PMID- 19293466 TI - Introducing improved voxel navigation and fictitious interaction tracking in GATE for enhanced efficiency. AB - Geant4 Application for Emission Tomography (GATE) is a widely used, well validated and very versatile application for Monte Carlo simulations in emission tomography. However, its computational performance is poor, especially for voxelized phantoms, partly due to the use of a very general particle tracking algorithm. In this work, two methods are proposed to reduce the time spent on particle tracking in the phantom: a newly introduced 'regular navigation algorithm' of Geant4 and fictitious interaction tracking (also known as Woodcock tracking) for photons. The speed-up introduced by the two methods was investigated by simulating a PET acquisition with the Allegro/GEMINI GXL PET/CT scanner. The simulation was based on a clinical head-and-neck [(18)F]FDG PET/CT scan. The total time spent for the simulation (including initialization, particle tracking and signal processing) was obtained for seven settings corresponding to different tracking options. All seven methods led to very close results with regard to the total number of detected coincidences (less than 0.5% differences), and trues, scatter and random fractions. Acceleration factors of approximately 2.7 (14 x 14 x 9 voxels) to 27.6 (378 x 378 x 243 voxels) were obtained in comparison with the fastest available tracking available in GATE 3.1.2. PMID- 19293467 TI - Scatter-based magnetic resonance elastography. AB - Elasticity is a sensitive measure of the microstructural constitution of soft biological tissues and increasingly used in diagnostic imaging. Magnetic resonance elastography (MRE) uniquely allows in vivo measurement of the shear elasticity of brain tissue. However, the spatial resolution of MRE is inherently limited as the transformation of shear wave patterns into elasticity maps requires the solution of inverse problems. Therefore, an MRE method is introduced that avoids inversion and instead exploits shear wave scattering at elastic interfaces between anatomical regions of different shear compliance. This compliance-weighted imaging (CWI) method can be used to evaluate the mechanical consistency of cerebral lesions or to measure relative stiffness differences between anatomical subregions of the brain. It is demonstrated that CWI-MRE is sensitive enough to reveal significant elasticity variations within inner brain parenchyma: the caudate nucleus (head) was stiffer than the lentiform nucleus and the thalamus by factors of 1.3 +/- 0.1 and 1.7 +/- 0.2, respectively (P < 0.001). CWI-MRE provides a unique method for characterizing brain tissue by identifying local stiffness variations. PMID- 19293468 TI - The angular dependence and effective point of measurement of a cylindrical scintillation dosimeter with and without a radio-opaque marker for brachytherapy. AB - Fibre optic scintillation dosimeters, consisting of a plastic scintillator coupled to an optical fibre, are a promising dosimeter in brachytherapy applications. The combination of tissue equivalence, real-time readout and small spatial size makes them especially attractive for in vivo verification of patient treatments. Given that the orientation of the dosimeter with respect to the radioactive source changes during brachytherapy treatment, the angular dependence of the dosimeter is important. We derived the dependence of the response of a cylindrical dosimeter to a point radiation source as a function of distance along its axis and along a radius. Using the results, the effective point of measurement of a cylindrical scintillator was located for two points in the angular response curve as a function of distance between the source and dosimeter. We measured the angular response experimentally for a cylindrical scintillation dosimeter, when the source was located at a distance of 50 mm from the centre of the scintillator. A refinement of the design, in which a radio opaque marker is incorporated into the tip for accurate localization in the patient, modifies the angular response of the dosimeter. For this new dosimeter design, we show that the dosimeter response decreases by 20% when the source is located on the axis of the scintillator, due to absorption by the marker. The dosimeter response becomes almost angle independent at 10 degrees away from the axis. Excluding this cone, a cylindrical scintillation dosimeter which incorporates a radio-opaque marker was found to be angle independent to within 2%. In most clinical brachytherapy applications, this design has an acceptable angular dependence. PMID- 19293469 TI - Occupational exposure measurements of static and pulsed gradient magnetic fields in the vicinity of MRI scanners. AB - Recent advances in magnetic resonance imaging (MRI) have increased occupational exposure to magnetic fields. In this study, we examined the assessment of occupational exposure to gradient magnetic fields and time-varying magnetic fields generated by motion in non-homogeneous static magnetic fields of MRI scanners. These magnetic field components can be measured simultaneously with an induction coil setup that detects the time rate of change of magnetic flux density (dB/dt). The setup developed was used to measure the field components around two MRI units (1 T open and 3 T conventional). The measured values can be compared with dB/dt reference levels derived from magnetic flux density reference levels given by the International Commission on Non-Ionizing Radiation Protection (ICNIRP). The measured motion-induced dB/dt values were above the dB/dt reference levels for both MRI units. The measured values for the gradient fields (echo planar imaging (EPI) and fast field echo (FFE) sequences) also exceeded the dB/dt reference levels in positions where the medical staff may have access during interventional procedures. The highest motion-induced dB/dt values were 0.7 T s( 1) for the 1 T scanner and 3 T s(-1) for the 3 T scanner when only the static field was present. Even higher values (6.5 T s(-1)) were measured for simultaneous exposure to motion-induced and gradient fields in the vicinity of the 3 T scanner. PMID- 19293470 TI - Optimizing functional parameter accuracy for breath-hold DCE-MRI of liver tumours. AB - Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) is a valuable tool for assessing treatment response to novel cancer therapeutics. With appropriate data acquisition, quantitative functional parameter estimates can be obtained by fitting a model to the data. This research focuses on applying a dual-input single-compartment pharmacokinetic model to breath-hold DCE-MRI imaging of the liver. In this paper, the use of two breath-holds, providing greater temporal information, is compared with a single breath-hold approach. Computer simulations are used to assess the accuracy, precision and sensitivity to input function errors obtained for parameters estimated from the two imaging protocols. Data from ten patients were analysed to assess the noise statistics obtained from the two breath-hold protocols. The noise statistics were used with a pharmacokinetic liver model to simulate data, from which the estimation accuracy, precision and sensitivity for the two protocols were assessed. Data from the ten patients were also analysed, and the estimates were compared with literature values. This work demonstrates the feasibility of obtaining functional liver perfusion estimates over a 3D volume using a sequential breath-hold protocol. The simulation results show that the protocol consisting of two images per breath-hold is to be preferred as it requires identical patient co-operation, but provides parameter estimates that have superior accuracy and precision. PMID- 19293471 TI - Development of a multi-layer ionization chamber for heavy-ion radiotherapy. AB - A multi-layer ionization chamber (MLIC) which consists of a stack of parallel plate-type ionization chambers in which the parallel configuration is in the depth direction was developed at the National Institute of Radiological Science (NIRS) and has been used as a field dosimeter for MU calibration since 2002. Although the MLIC can measure depth dose distributions at one time, a correction is needed to obtain an accurate dose at the center of the spread-out Bragg peak (SOBP) in a water phantom. We attributed the observed difference between the correct dose at the center of the SOBP and the measured dose to the lack of water equivalence of the MLIC. In order to overcome this problem, a new MLIC was developed. It consists of polymethyl-methacrylate (PMMA) plates and graphite electrodes instead of flame retardant type 4 (FR4) and copper electrodes. The calibration coefficients of the MLICs were obtained by 160 MeV proton beam irradiation. For carbon-ion beams, the PMMA type MLIC has the capability to measure depth dose distributions in the water phantom with less than 2% error, including the fragment tail region. PMID- 19293472 TI - The efficiency of CD40 down regulation by siRNA and antisense ODN: comparison of lipofectamine and FuGENE6. AB - BACKGROUND: Dendritic cells (DCs) are ideal accessory cells in the field of gene therapy. Delivery of DNA and siRNA into mammalian cells is a useful technique in treating various diseases caused by single gene defects. Selective gene silencing by small interfering RNAs (siRNAs) and antisense oligodeoxynucleotides (ODN)s is an efficient method for the manipulation of cellular functions. An efficient, functional delivery system with no toxicity problems would be attractive. OBJECTIVE: We compared two commercially available cationic lipids, Lipofectamine and FuGENE6, in the delivery of both siRNA and antisense ODNs into mice spleen derived DCs. METHODS: Cellular uptake was measured by the means of fluorescein labelled non-silencing siRNA and antisense ODNs as a model system using flow cytometry. Cytotoxicity of the two delivery systems was compared with propidium iodide and annexin-V staining, and quantified with flow cytometry. The efficiency of our oligonucleotide delivery systems was compared by measuring CD40 expression by flow cytometry. RESULTS: CD40 expression in DCs was 38%. After siRNA transfection by Lipofectamine, CD40 expression decreased to 13%, and after transfection by FuGENE6, it decreased to 18%. The difference was statistically significant. CD40 down regulation in DCs transfected with the two different antisense sequences by Lipofectamine was 21% and 23%, and down regulation after transfection by FuGENE6 was 19% and 18%, respectively. The differences were not statistically significant. The effects of siRNA and antisense ODNs were specific. CONCLUSION: Lipofectamine was a more potent delivery system in siRNA effect, followed by FuGENE6. There was no significant difference between Lipofectamine and FuGENE6 as a delivery system of antisense ODNs. PMID- 19293473 TI - Immunogencity of HSA-L7/L12 (Brucella abortus ribosomal protein) in an animal model. AB - BACKGROUND: The immunogenic Brucella abortus ribosomal protein L7/L12 is a promising candidate antigen for the development of subunit vaccines against brucellosis. OBJECTIVE: This study was aimed to evaluate the protection of recombinant Human Serum Albumin (HAS)-L7/L12 fusion protein in Balb/c mice. METHODS: The amplified L7/L12 gene was cloned in pYHSA5 vector, pYHSA5-L7/L12 construct was transformed in Saccharomyces cerevisiae and the expressed protein from supernatant was purified by affinity chromatography. Balb/c mice were immunized in five groups by tHSA-L7/L12 fusion protein (group 1), Brucella abortus S19 (group 2), HSA (group 3), recombinant L7/L12 (group 4), PBS (group 5). ELISA to detect antibody production, LTT test to assess antigen specific lymphocyte response were conducted prior to virulent B. abortus strain 544 challenge two weeks after the last injection. Bacterial counts from spleens of immunized mice were done four weeks after challenge. RESULTS: In ELISA tests, the specific antibodies exhibited a dominance of immunoglobulin IgG1 over IgG2a. In addition, the tHSA-L7/L12 fusion protein and L7/L12 elicited a strong T-cell proliferative response upon restimulation in vitro with recombinant tHSA-L7/L12 and L7/L12, suggesting the induction of a cellular immunity response in vivo. However, there was no significant difference in proliferative response of L7/L12 and tHSA-L7/L12 fusion protein (p>0.05). The L7/L12 and tHSA-L7/L12 fusion protein vaccines could also induce significant protection against challenge with the virulent strain B. abortus 544 in Balb/c mice (p< or =0.05). CONCLUSION: The tHSA-L7/L12 fusion protein, similar to L7/L12 has the ability to induce antigen specific lymphocyte proliferation, stimulate humoral immunity and engender protection. PMID- 19293474 TI - Kinetics of primary and memory cytotoxic T lymphocyte responses to herpes simplex virus 1 infection: granzyme B mediated CTL activity. AB - BACKGROUND: Herpes simplex virus type 1 is one of the most common viruses among human population. Studies demonstrate the essential role of cell mediated immunity, especially CD8+ T cells, in prevention and clearance of HSV1. OBJECTIVE: It is of great importance to improve our knowledge about the kinetics of CTL responses to primary and secondary HSV-1 infection. METHODS: Using a sensitive technique for detection and analysis of CD8+ T cells, granzyme B ELISA, the CTL activity in the spleens of Balb/c mice at various time points after intraperitoneal administration of HSV1 (strain KOS) in primary and secondary infections were determined. RESULTS: During acute HSV-1 infection, virus specific cytotoxic T cells were detected at day 5 post virus inoculation and peaked at day 7. Six hours after secondary infection the activity of memory CD8+ T cells was detected and peaked at 12 hours post infection. CONCLUSION: The peak of CTL activity was found to be day 7 post infection in primary HSV-1 infections which decreased with time. In secondary infections, the activity of CTLs reached the highest level at 12 hours post infection. PMID- 19293475 TI - Effect of Fas -670 A/G gene polymorphism on corneal allograft endothelial rejection. AB - BACKGROUND: Human cornea expresses functional Fas-ligand capable of killing Fas+ activated lymphocytes. Fas expression is partly regulated by -670 A/G polymorphism in the promoter region of Fas gene. OBJECTIVE: The aim of the present study is to determine the association between Fas-670A/G polymorphism and survival of corneal transplantation. METHODS: In 276 graft recipients who mainly underwent penetrating keratoplasty because of keratoconus, bullous keratopathy and corneal opacity, Fas -670 A/G polymorphism was determined by allele specific oligonucleotide polymerase chain reaction (ASO-PCR) techniques. RESULTS: There was no statistically significant relationship between Fas -670 A/G polymorphism and rejection episode (p=0.35). Moreover, the relationship between this polymorphism and rejection episode outcome (transplant recovery vs failure) was not statistically significant (p=0.13). CONCLUSION: The results of the present study show no significant correlation between corneal graft rejection, rejection recovery and Fas -670A/G gene polymorphism. PMID- 19293476 TI - The influence of iron loading and iron chelation on the proliferation and telomerase activity of human peripheral blood mononuclear cells. AB - BACKGROUND: Iron is an essential trace element in cell proliferation. Several investigations demonstrate that iron deprivation inhibits cell proliferation. However, the impact of iron on telomerase activity of activated lymphocytes remains unexplained to date. OBJECTIVE: In this study, the effect of iron on the proliferation and telomerase activity of lymphocytes stimulated by phytohemagglutinin (PHA) were investigated. METHODS: Iron loading was performed by incubating peripheral blood mononuclear cells in 500microM FeSO4.7H2O for 24 h and iron chelation was done by exposing cells to desferrioxamine, a potent iron chelator. The effects of silymarin, a flavonoid with both antioxidant and iron chelating activities, on the proliferation and telomerase activity of PHA activated lymphocytes were also compared with desferrioxamine. Proliferation and telomerase activity were assessed using BrdU incorporation assay and Telomeric Repeat Amplification Protocol (TRAP), respectively. RESULTS: The proliferations of lymphocytes were significantly inhibited by 10 and 20 microg/ml desferrioxamine in a dose dependent manner, while iron loading recovered suppressed cell proliferation to the normal level. Silymarin at 20 microg/ml significantly increased the proliferation of lymphocytes in both normal and iron treated conditions. Telomerase activity of lymphocytes was markedly increased by iron treatment and suppressed by desferrioxamine. Conversely, iron treatment had no effect on the telomerase activity of lymphocytes incubated with silymarin. CONCLUSION: Iron plays a significant role in the proliferation and telomerase activity of lymphocytes. The effects of silymarin on the proliferation and telomerase activity of lymphocytes were completely different from those of desferrioxamine, suggesting that the immunomodulatory effect of silymarin is probably not associated with its iron chelating activity. PMID- 19293477 TI - Age specific reference levels of serum prostate-specific antigen, prostate volume and prostate specific antigen density in healthy Iranian men. AB - BACKGROUND: It is relevant to highlight that there is not a precise and perfect report on either 95 percentile value (upper limit of normal range) or on appropriate reference intervals for serum PSA in Iranian population. OBJECTIVE: To determine age-specific reference ranges for serum prostate-specific antigen (PSA) concentration and PSA density (PSAD) and prostate volumes in a population of healthy Iranian men. METHODS: Nine-hundred and thirteen healthy Iranian men, aged 50-79 years, underwent a detailed clinical evaluation including a digital rectal examination, a serum PSA determination (DRE) and transrectal ultrasound (TRUS). PSA test was performed on 666 of the subjects and TRUS was done on 633 of them. None of the subjects had any evidence of prostate cancer by any one of the three diagnostic tests and had no history of Lower Urinary Tract Symptoms (LUTS). Age specific ranges for PSA levels, PSA density and prostate volume were determined. RESULTS: The serum PSA concentration correlated directly with the subjects' age (r=0.280; p<0.001) and prostatic volume (r=0.327; p<0.001). Also prostatic volume was directly proportional to age (r=0.197; p<0.001).The serum PSA ranges (95th percentile) for each age range in Iranian men were: 0.00-2.61 ng/ml for 50-59 years; 0.00-3.59 ng/ml for 60-69 years; and 0.00-4.83 ng/ml for 70-79 years. The respective prostate volumes were: 14-59, 16-66 and 18-73ml. Also respective PSA densities were: 0.00-0.076, 0.00-0.10 and 0.00-0.14 ng/ml/ml. CONCLUSION: The present study confirms earlier reports that serum PSA levels and prostate volume and PSAD are age- and race- dependent, so it is appropriate to have age- specific reference ranges for these variables in various communities around the world. This will increase the positive predictive value of PSA estimation in the diagnosis of prostate cancer in different communities. PMID- 19293478 TI - Distribution of HLA-B*27 alleles in patients with ankylosing spondylitis in Iran. AB - BACKGROUND: HLA-B*27 is strongly associated with ankylosing spondylitis (AS). It represents a family of alleles that differ among ethnic groups. OBJECTIVE: The aim of this study was to determine the distribution of HLA-B*27 alleles in AS patients and healthy controls in Isfahan (Iran). METHODS: Sixty AS patients and 430 healthy blood donors were selected. All subjects were HLA-B*27 positive by flow cytometry. HLA-B*27 subtypes were determined by PCR-SSP. RESULTS: Forty patients (66.7%) and 17 controls (3.95%) were HLA-B*27 positive. Subtypes detected by PCR-SSP were B*2705, B*2702, B*2704 and B*2707. One patient was B*2702/B*2710. No significant difference was found in the distribution of these alleles between AS patients and controls. CONCLUSION: Although Caucasian subtypes are predominant among Iranians, this population is characterized by a combination of both specific Caucasian and Oriental subtypes. However such results should be interpreted carefully because of the small sample size in our investigation and definitive conclusion awaits more ethnic-group studies. PMID- 19293479 TI - Successful treatment of refractory Behcet's disease with the TNF-alpha blocker infliximab. PMID- 19293480 TI - Inverse planning for the T1--T2 conundrum in translation research. PMID- 19293481 TI - Concurrent chemoradiotherapy in the management of advanced nasopharyngeal carcinoma: current status. AB - Nasopharyngeal carcinoma (NPC) differs from other head and neck cancers in its epidemiology, natural behavior and, particularly, in the therapeutic considerations. In addition, NPC is responsive to both chemotherapy and radiotherapy (RT). This article reviews the recent advances in concurrent chemoradiotherapy (CCRT) for NPC. To identify the studies mentioned in this article, we searched the MEDLINE database, and abstracts of selected conference proceedings (up to 2008) using the key words 'nasopharyngeal carcinoma,' 'advanced,' 'radiotherapy,' and 'concurrent chemotherapy.' Eight randomized clinical trials of CCRT in NPC have been reported in the English literature. These trials can be classified according to the timing of the chemotherapy, which was given either concurrently with RT (in three trials) or concurrently with radiotherapy and adjuvant after the and of concurrent chemoradiotherapy (in five trials). There have also been four meta-analyses addressing the value and scheduling of chemotherapy in the curative treatment of NPC. This article reviews the recent literature and the pertinent issues concerning the role of CCRT in the treatment of patients with locoregionally advanced NPC. PMID- 19293482 TI - Long-term results of LINAC-based stereotactic radiosurgery for acoustic neuroma: the Greek experience. AB - PURPOSE: To estimate the value of LINAC-based stereotactic radiosurgery (SRS) for the long-term local control of unilateral acoustic neuromas. MATERIALS AND METHODS: Twenty patients (median age 66; range 57-80 years) with unilateral acoustic neuroma underwent LINAC-based SRS from May 2000 through June 2004 with a dose of 11-12 Gy. The follow-up period ranged from 36 to 84 months (median follow up period: 55 months). Before SRS none of the patients had useful hearing. The follow-up consisted of repeat imaging studies and clinical examination for assessment of facial and trigeminal nerve function at 6-month intervals for the first year and yearly thereafter. RESULTS: Eleven tumors (58%) decreased in size and eight (42%) remained stable. One tumor showed a minor increase in size on the MRI done 6 months after SRS in comparison with the pretreatment MRI; however, a subsequent decrease was noticed on the next radiographic assessment and the tumor remained stable from then on. None of the tumors increased in size in the long term follow-up, thus giving an overall growth control of 100% for the patients in this study. None of the patients had useful hearing before SRS, so hearing level was not assessed during follow-up. No patient developed new, permanent facial or trigeminal neuropathy. CONCLUSION: LINAC-based SRS with 11-12 Gy provides excellent tumor control in acoustic neuroma and has low toxicity even after long term follow-up. PMID- 19293483 TI - Chondrosarcoma of bone: does the size of the tumor, the presence of a pathologic fracture, or prior intervention have an impact on local control and survival? AB - AIMS AND OBJECTIVES: We studied 45 patients with chondrosarcoma, without metastasis at diagnosis, who were surgically treated between January 2000 and December 2004 to evaluate the risk factors associated with local recurrence and metastasis. MATERIALS AND METHODS: Fourteen (31%) patients had had some major prior intervention, either in the form of an open biopsy or a curettage / unplanned excision, before presenting to us. Eight patients had pathologic fractures at presentation. None of the patients received adjuvant chemotherapy or radiotherapy. The follow-up duration ranged from 8-75 months. All survivors had a minimum follow-up of 36 months (range 36-75 months). RESULTS: There were 11 grade 1 (24.5%), 23 grade 2 (51%), and 11 grade 3 (24.5%) chondrosarcomas. Thirty-two (71%) patients had tumors that were larger than 8 cm in the greatest dimension. Margins were adequate in 31 patients. Twenty-five patients had disease relapse; there were four local failures, nine distant failures, and 12 combined failures. At the time of the last review, 12 patients had died, 11 were alive with disease, and 22 were free of disease. The cumulative event-free survival was 44% and the overall survival was 73%. CONCLUSION: Grade of tumor, size of tumor, and adequacy of resection might be important predictors of outcome. Local recurrence is a prelude to distant metastasis and portends poor ultimate survival. The presence of a pathological fracture could indicate biologically aggressive disease, and limb salvage in these cases should be advised with caution. Even in cases where there has been a prior unplanned intervention, local control can be achieved by subsequent adequate resection. PMID- 19293484 TI - Field-in-field technique for upper abdominal malignancies in clinical radiotherapy. AB - BACKGROUND: In upper abdominal malignancies (UAM), due to the presence of multiple inhomogeneous tissues, a wedge-based conformal treatment planning results in high-dose regions inside the target volume. AIM: This study was designed to explore the feasibility of using a field-in-field (FIF) technique in different UAM and its efficacy in reducing the high-dose regions. MATERIALS AND METHODS: Twelve patients of UAM (which included malignancies of the gastroesophageal junction, stomach, gall bladder, and pancreas) were selected for this study. Computed tomography (CT) scans were performed and three-dimensional conformal wedge plans were generated for all the cases. The same plan was copied with the wedges removed and a FIF plan was generated. The two plans were compared for mean, maximum, and median doses; dose received by 2% (D2) and 98% (D98) of the target volume; volume receiving> 107% (V > 107%) and < 95% (V Statistical Analysis: Statistical analysis was performed with SPSS, version 10.0. RESULTS: For all the cases, the FIF technique was better than wedge-based planning in terms of maximum dose, D2, V > 107%, and CI; there was a statistically significant reduction in monitor units. With regard to doses to critical structures, there was marginal dose reduction for the kidneys and spinal cord with FIF as compared to wedge-based planning. CONCLUSION: The FIF technique can be employed for UAM in place of wedge-based conformal treatment plans. PMID- 19293485 TI - Reirradiation with high-dose-rate remote afterloading brachytherapy implant in patients with locally recurrent or residual cervical carcinoma. AB - PURPOSE: To assess the outcome for patients with postradiation locally recurrent or residual cervical malignancies treated with high-dose-rate (HDR) remote afterloading brachytherapy implant. This is a prospective study. MATERIALS AND METHODS: Twenty-two patients with postradiation recurrent cervical carcinomas were treated with HDR interstitial brachytherapy. All of these 22 patients had received some form of radiotherapy prior to implant placement and had biopsy proven recurrence. Implant dose for these patients was in the range of 12-45 Gy, with a median dose of 25.80 Gy. RESULTS: Overall survival was better for patients in the 'no bad prognostic factor' (NBPF) group than for patients in the 'bad prognostic factor' (BPF) group and this difference was statistically significant. Overall survival was better for patients who had stage II disease before implant placement than for those who had stage III disease and also for patients who had relatively less parametrial disease. Grade 4 complications were seen only in four patients and were within acceptable limits. Multivariate analysis showed that only the BPF variable had a statistically significant effect on survival. CONCLUSION: We conclude that reirradiation with HDR brachytherapy implant in recurrent or residual carcinoma cervix can give a good chance of survival, with acceptable morbidity, if proper selection of patients is done. This treatment should not be done for palliative purposes. PMID- 19293486 TI - Comparison between young and old patients with bronchogenic carcinoma. AB - OBJECTIVE: This study was undertaken to explore the clinicopathological profile of bronchogenic carcinoma in young patients. MATERIALS AND METHODS: The present study was conducted on 799 consecutive histopathologically proven cases of bronchogenic carcinoma that were referred from different parts of Uttar Pradesh. RESULTS: Out of 799 patients, 73 patients (9.1%; 59 males and 14 females) were < or = 40 years of age and were classified as 'young' patients. The mean ages of the subjects in the younger and older patient groups were 36 and 58 years, respectively. Among the older patients, 590 (81.3%) were smokers, and there were 53 (72.6%) smokers among the younger patients. Squamous cell carcinoma was the commonest histological subtype in both the groups, but squamous cell carcinoma was more frequently diagnosed in older patients than in younger patients. CONCLUSION: This study suggests that, regardless of age or sex, lung cancer must be ruled out in all patients who have persistent signs of pulmonary disease and a history of heavy smoking. PMID- 19293487 TI - Importance of contouring the cervical spine levels in initial intensity-modulated radiation therapy radiation for head and neck cancers: implications for re irradiation. AB - AIM: To evaluate the maximum differential cervical spinal (C-spine) cord dose in intensity-modulated radiation therapy (IMRT) plans of patients undergoing radiotherapy for treatment of head and neck cancer. MATERIALS AND METHODS: The C spine of ten head and neck cancer patients that were planned using IMRT and each cervical vertebral body and the right and left sides was contoured by splitting the cord in the center. Dose-volume histograms (DVH) and maximum point doses were obtained for each contour and compared. RESULTS: The dose to the cord varied with the location of the primary tumor but such variation was not consistently seen. This report provides information that is critical for planning reirradiation treatments. We recommend that contouring of the C-spine cord with IMRT should include outlining of each cervical cord level and identification of the right and the left sides of the cord on each plan. PMID- 19293488 TI - Invasive thymoma with intraorbital metastases. AB - Thymomas are epithelial neoplasm of thymus and most common primary neoplasm of anterior and superior mediastinum affecting males and females equally. It occurs usually in the fifth to seventh decade. Approximately one-third of thymomas are invasive. Metastases to distant extrathoracic sites such as the liver, lung, lymph node, kidneys, ovary and brain occur infrequently. This is more common with invasive thymomas. Although brain has been shown to be a site of infrequent metastases, intraorbital metastases has not yet been reported. Here we report one such case of invasive thymoma of anterior superior mediastinum, which later metastasized to orbit as well as the brain. PMID- 19293489 TI - Butterfly glioma of the corpus callosum. AB - The prognosis of glioblastoma multiforme (GBM) is poor even with aggressive first line therapy, which includes surgery, radiation therapy, and adjuvant chemotherapy. Although the ideal course of treatment for elderly patients with newly diagnosed GBM is still undecided and requires further studies, the new chemotherapeutic agents administered with or without concomitant radiation therapy have shown promising results. However, in our setting, where resources are limited and newer treatment options are expensive, it is often difficult to deliver the best care to the patient. PMID- 19293490 TI - Hypercalcemia and treated breast cancers: the diagnostic dilemma. AB - The relationship of hypercalcemia with cancer is well described in the literature. Breast cancer is the commonest malignancy associated with hypercalcemia; the detection of hypercalcemia in these patients usually signifies metastatic disease and is associated with a poor prognosis. However, the treating oncologist should keep in mind that a strong correlation exists between breast cancer and primary hyperparathyroidism. We present a case of a patient of treated breast cancer who, in the absence of metastatic bone disease, developed hypercalcemia due to hyperparathyroidism secondary to a parathyroid adenoma. PMID- 19293491 TI - Numb chin syndrome as a manifestation of metastatic squamous cell carcinoma of esophagus. AB - Numb chin syndrome (NCS) is a sensory neuropathy presenting with numbness of the chin in the distribution of the mental nerve and the branches of the mandibular division of the trigeminal nerve. Though it can be caused by a benign process, NCS should be regarded as being due to malignancy until proven otherwise. Among the malignancies that cause NCS the most common are breast cancer, prostate cancer, and lymphoreticular malignancy. In squamous cell carcinoma (SCC) of the esophagus, spread to the mandible is a rare and often late event. An often overlooked clinical sign in mandibular metastases is hypoesthesia or paresthesia over the peripheral distribution of the inferior alveolar nerve/mental nerve; this sign has been referred to in the literature as NCS or numb lip syndrome or mental nerve neuropathy. Rarely, this may be the first presentation of a disseminated malignancy. Prognosis is usually poor. The discovery of this symptom should alert the clinician to the possibility of disseminated disease. In this article we report a rare case of metastatic SCC of the esophagus in a 40-year-old male patient who presented with NCS. We also review the mechanism, causes, and evaluation of NCS. PMID- 19293492 TI - Primary non-Hodgkin lymphoma of bone: an unusual presentation. AB - Primary lymphoma of bone (PLB) is an extremely rare condition that is usually confused with other primary injuries of the bone. It is characterized by the involvement of one or more bone locations, with or without involvement of regional lymph nodes and viscera. PLB constitutes 3-7% of all malignant bone tumors and approximately 3% of all extranodal lymphomas. It is found at all ages, being most frequently seen in adult life. Any part of the skeleton can be involved, but a trend exists in favor of bones with persistent bone marrow. We report a case of PLB with an unusual presentation: involvement of the proximal phalanx of the thumb. Treatment with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) plus etoposide established complete remission. Consolidation with radiotherapy of the femur and phalanx was performed. There was no evidence of recurrence at the 14 th month follow-up. PMID- 19293493 TI - Rhabdoid variant of lung cancer: clinicopathological details of a case and a review of literature. AB - Primary rhabdoid tumor of lung is a rare histological and clinical entity. Lung tumors with rhabdoid features have been included as variants of large-cell carcinoma in the 1999 World Health Organization (WHO) classification of lung tumors. A large-cell carcinoma with a rhabdoid phenotype (LCCRP) is unusual, with only 38 cases reported till date. We report the clinical details of one such case that was treated with pneumonectomy and adjuvant chemotherapy. We also present a review of the literature. To identify relevant articles, we searched PubMed, Ovid, and IngentaConnect databases using the key words 'rhabdoid,' 'lung cancer,' and 'primary rhabdoid tumor of lung.' PMID- 19293494 TI - The enigma of proton therapy for head and neck cancers. PMID- 19293495 TI - Leprosy: a disease with diagnostic and management challenges! PMID- 19293496 TI - Vitiligo and diet: a theoretical molecular approach with practical implications. PMID- 19293497 TI - Targeted phototherapy. AB - Phototherapy is one of the most important therapeutic modalities in dermatology. This field has seen several major advances in the recent years, the most recent being targeted phototherapy. Targeted phototherapy, which includes laser and nonlaser technologies, delivers light/laser in the ultraviolet spectrum, of specific wavelength, specifically targeted at the affected skin and thereby avoids many of the side effects of conventional phototherapy. The treatment has been claimed to be effective, quick, and needing fewer treatment sessions. The article reviews this new mode of phototherapy. PMID- 19293498 TI - Relapse in leprosy. AB - Leprosy is unique in terms of the nature of the causative organism (Mycobacterium leprae), the chronicity of the disease, its prolonged treatment and the definitions of "cure" and "relapse." The principal mode of assessing the efficacy of therapeutic regimens in leprosy is the "relapse rate." There are wide variations in estimates of relapse rates after the World Health Organization (WHO) multidrug therapy in different regions. The important predisposing factors for relapse include the presence of "persister" bacilli, monotherapy, inadequate/irregular therapy, presence of multiple skin lesions/thickened nerves and lepromin negativity. The conventional methods of confirming activity or relapse in an infectious disease (demonstration and/or culture of the etiologic agent) have limited utility in leprosy because of the difficulty in demonstrating bacilli in paucibacillary (PB) cases and absence of a method of in vitro cultivation of M. leprae. Bacteriological parameters are useful in multibacillary (MB) leprosy, whereas in PB leprosy, the criteria for relapse depend primarily on clinical features. Although there are no widely available serologic tests for leprosy other than in a research setting, various immunological tests may be useful for monitoring patients on chemotherapy as well as for confirming suspected cases of relapse. The main differential diagnoses for relapse are reversal reactions, erythema nodosum leprosum and reactivation/resistance/reinfection. The most reliable criteria for making an accurate diagnosis of relapse include clinical, bacteriological and therapeutic criteria. Additional ones that may be used, depending on the setting, are histopathological and serologic criteria. Relapsed cases of leprosy should be identified and put back on chemotherapy as soon as possible to prevent further disability and transmission of infection. Factors that should be considered in choosing an appropriate regimen are the type of leprosy (PB or MB), previous treatment and drug resistance. Occasionally, clinicians may need to use their judgement to modify the standard WHO treatment regimens according to the scenario in each patient. PMID- 19293499 TI - Contact sensitization in venous eczema: preliminary results of patch testing with Indian standard series and topical medicaments. AB - BACKGROUND AND AIMS: Elimination of allergens/topical medications causing contact dermatitis in venous eczema, which poses a significant problem in its chronicity and treatment, provides the basis for better therapeutic outcome. Our objective was to determine the pattern of contact sensitization in venous eczema patients in Himachal Pradesh (India). METHODS: Thirty-four patients (M:F, 31:3) and 10 controls (M:F, 6:4) were patch tested with Indian standard series and 10 commonly used topical medicaments. RESULTS: Positive patch test results were seen in 50% (M:F, 16:1) of the patients. Common allergens were Fragrance mix (15%), p phenylendiamine (15%), nickel (9%), wool alcohol (9%), chinoform (9%), balsum of Peru (5%), cobalt chloride (5%), potassium dichromate (3%), epoxy resin (3%), thiuram mix (3%) and formaldehyde (3%). Only sisomycin and miconazole among the topical medications elicited a positive patch test reaction in 3 and 5% patients, respectively. Neomycin contact sensitivity was not seen in any of the patients. One patient who had exacerbation of venous eczema following accidental application of topical diclofenac showed a positive patch test reaction to it. CONCLUSIONS: Patch test should be used to identify the topical agents that may be responsible for perpetuation or aggravation of eczema, especially in patients who do not improve despite adequate treatment of other underlying cause(s). PMID- 19293500 TI - Epidemiological survey of dermatophytosis in Tehran, Iran, from 2000 to 2005. AB - BACKGROUND: Cutaneous fungal infections are common in Tehran, Iran, and causative organisms include dermatophytes, yeasts and non-dermatophyte molds. The prevalence of superficial mycosis infections has risen to such a level that skin mycoses now affect more than 20-25% of the world's population, making them the most frequent form of infection. AIMS: Our aim was to determine the prevalence of superficial cutaneous fungal infections especially dermatophytosis in our Medical Mycology Laboratory in the Pasteur Institute of Iran, Tehran. METHODS: A total of 17,573 specimens were collected from clinically suspected tinea corporis, tinea cruris, tinea capitis, tinea faciei, tinea pedis, tinea manuum and finger and toe onychomycosis from 2000 to 2005. Patients were referred to our laboratory for direct examination, fungal culture and identification. The incidence of each species was thus calculated. RESULTS: Dermatophytes remain the most commonly isolated fungal organisms, except from clinically suspected finger onychomycosis, in which case Candida species comprise >7% of the isolates. Epidermophyton floccosum remains the most prevalent fungal pathogen and increased incidence of this species was observed in tinea cruris. Trichophyton tonsurans continues to increase in incidence. CONCLUSION: This study identifies the epidemiologic trends and the predominant organisms causing dermatophytosis in Tehran, Iran. These data can be used to ascertain the past and present trends in incidence, predict the adequacy of our current pharmacologic repertoire and provide insight into future developments. Consideration of the current epidemiologic trends in the incidence of cutaneous fungal pathogens is of key importance to investigational effort, diagnosis and treatment. PMID- 19293501 TI - Ocular abnormalities in atopic dermatitis in Indian patients. AB - BACKGROUND AND AIMS: Atopic dermatitis (AD) is a common skin disease. Long standing, severe AD with repeated scratching and rubbing of the face, which requires continuous dermatologic care, predisposes the patient to various ocular complications. The knowledge of the frequency and significance of these ocular complications may allow their early diagnosis and treatment. The present study assesses the ocular complications in Indian children suffering from AD. METHODS: In order to study the ocular complications in AD, 100 patients (61 male and 39 female) between the ages of 1 and 14 years were recruited. All the patients had complete dilated fundus examination with indirect ophthalmoscopy. The lid, conjunctiva and cornea were examined. Also, any evidence of cataract formation and retinal disorders were recorded. RESULTS: The mean age of the children was 5.4 years. Forty-three (43.0%) AD patients showed ocular abnormalities in the form of lid and conjunctival changes. Of these, 18 (41.9%) patients showed only lid involvement, 16 (37.2%) only conjunctival involvement and both conjunctival and lid changes were seen in nine (20.9%) patients. Conjunctival changes were mostly in the form of a cobblestone appearance of the papillae, with mild to moderate papillary reaction and papillary hypertrophy. Variables observed to have a significant impact on the development of ocular abnormalities were age more than 5 years, duration of illness >12 months, positive family history of atopy, presence of palmar hyperlinearity and a combination of both xerosis and Dennie Morgan fold. CONCLUSIONS: The present study is the first of its kind from India to document an association between AD in children and various ocular manifestations. The ocular manifestations observed in our cohort were not associated with significant ocular morbidity or visual impairment possibly because of a less-severe disease in Indians. PMID- 19293502 TI - Analysis on value of household contact survey in case detection of leprosy at a low endemic situation in China. AB - BACKGROUND: Leprosy is at a low endemic situation in China, the value of house contact survey in case detection of leprosy becoming a dispute. AIMS: To evaluate the value of household contact survey in the case detection of leprosy at a low endemic situation in China. METHODS: A study was carried out using a designed questionnaire in a retrospective method to analyze the value of household contact survey in case detection in Southwest and East China. RESULTS: A total of 2135 index leprosy patients were collected from January 1, 1996 to December 31, 2005 in six provinces of China. The number of index patients accounted for 22.0 and 14.1% of newly registered patients in the Southwest and East of China, respectively. The household contact survey (36.1%) and the skin clinic (62.0%) ranked first in methods of case detection in Southwest and East China, respectively. Within 5 years after primary leprosy patients were detected, 24.8 and 16.1% of the index patients in Southwest China and East China had been detected, respectively. CONCLUSION: The authors conclude that at the time of a low leprosy endemic situation, the household contact survey is still a useful method for case detection in China. PMID- 19293503 TI - Electronic medical records in dermatology: practical implications. AB - BACKGROUND: Electronic medical records (EMRs) can be of great use in dermatological data recording. Unfortunately, not many studies have been carried out in this specific area. AIMS: We attempt to evaluate the use of an EMR system in dermatology, comparing it with a conventional paper-based system. METHODS: Two hundred patient records of patients attending the dermatology outpatient department were studied over a 3-month period. Half the reports were entered in the conventional paper-based format while the other half was entered in an EMR system. The time taken for each consultation was recorded and the same was carried out for the first subsequent follow-up visit. RESULTS: The average time taken for the completion of the EMR-based consultation for new cases was 19.15 min (range, 10-30 min; standard deviation, 6.47). The paper-based consultation had an average time of 15.70 min (range, 5-25 min; standard deviation, 6.78). The P-value (T-test was used) was 0.002, which was significant. The average time taken for consultations and entering progress notes in the follow-up cases was slightly less than 10 min (9.7) for EMR while it was slightly more than 10 min (10.3) for the paper format. The difference was not statistically significant. The doctors involved also mentioned what they felt were the advantages and disadvantages of the system along with suggestions for improvement. CONCLUSION: The use of an EMR system in dermatology (or for that matter in any specialty) may overawe most users at the beginning, but once a comfort level is established, EMR is likely to outscore conventional paper recording systems. More time-motion-case studies are required to ascertain the optimal usage of EMR systems. PMID- 19293504 TI - Evaluation of narrow-band UVB phototherapy in 150 patients with vitiligo. AB - BACKGROUND: Very few studies have been performed to evaluate the efficacy and safety of narrow-band ultraviolet B (NBUVB) therapy in Indian patients with vitiligo and are of small sample size. AIMS: The purpose of this study is to know the efficacy and safety of NBUVB in 150 vitiligo patients of various age groups. METHODS: One hundred fifty patients (69 males, 81 females), aged 3-70 years, with vitiligo were treated twice weekly with NBUVB. The starting dose was 250 mJ/cm2 in adults and 150 mJ/cm2 in children, with 20% dose increments at each subsequent visit given for a maximum period of 1 year and were followed-up for 6 months for stability of repigmentation. Statistical methods were employed to establish the relation between the response and the number of exposures, duration of treatment, cumulative dose and the compliance. RESULTS: Analysis of our study showed that a majority of our cases, about 73, achieved 25-75% repigmentation, with an average of 51+/-19 exposures, 51 had <25% repigmentation, with an average of 19+/-11 exposures and 26 had >75% repigmentation, with an average of 74+/-24 exposures. Good response to therapy was directly associated with good compliance, more number of exposures and increasing cumulative dose, which was statistically significant (P<0.01). Adverse effects were minimal. Only three patients developed depigmentation of repigmented sites during follow-up. CONCLUSION: Our study proves that NBUVB therapy is an effective and safe tool in the management of vitiligo, with good stability of repigmentation and cosmetic appearance. PMID- 19293505 TI - Spitz nevus of the genital mucosa. AB - We herein report an 11-year-old girl who came to our clinic with a swelling on the genital area of 2 months duration. Dermatological examination of the patient was performed and a pigmented lesion was found on the inner surface of the labium majus of the mucosa. The lesion was well circumscribed and approximately 1 cm in diameter, with homogenous color distribution. The patient was diagnosed as Spitz nevus on the basis of clinical and histopathological findings. Our case is probably the first reported case of Spitz nevus localized to the genital mucosa in the English literature. PMID- 19293506 TI - Syringocystadenoma papilliferum on the thigh: an unusual location. AB - A 28-year-old man presented with a partially eroded growth on the back of his right thigh for the last 15 years. Tuberculosis verrucosa cutis and pyogenic granuloma were suspected on clinical examination. Histopathological examination revealed syringocystadenoma papilliferum (SCAP). SCAP is rare on the thigh and a review of the English literature revealed eight cases. Non-descript clinical presentation in an unusual location and the rare linear and segmental variants lead to misdiagnosis. Onset at puberty should alert one to the possibility of the unusual location of SCAP. PMID- 19293507 TI - Two unusual cases of histoplasmosis in human immunodeficiency virus-infected individuals. AB - Histoplasmosis, also called as Darling's disease, is caused by the dimorphic fungus, Histoplasma capsulatum. In India, several cases of histoplasmosis have been reported since 1954, but in only a few cases could the diagnosis be confirmed by fungal culture. Disseminated histoplasmosis in adults is often associated with immunosuppression, as in human immunodeficiency virus (HIV) infection. Oral lesions are seen in 30-50% of the patients. Here, we are reporting two histoplasmosis cases, one with disseminated histoplasmosis with extensive skin and oral lesions in a known HIV-positive patient and the second case presenting with ulcer of the tongue, found to be HIV positive on investigation. It is important to consider histoplasmosis as one of the differential diagnosis of oral lesions in HIV-infected individuals. PMID- 19293508 TI - Relapse of leprosy presenting as nodular lymph node swelling. AB - Lymphadenopathy is known to be associated with lepromatous leprosy and has also been observed as a feature of type-2 lepra reaction. However, nodular lymph node enlargement is not commonly reported in leprosy patients or as a feature of relapse. We herewith are presenting a case of bacteriological relapse in a patient of lepromatous leprosy treated 22 years before till smear negativity with WHO multidrug therapy (MDT) multibacillary type (MB). She presented with prominent nodular swelling of the cervical group of lymph nodes along with generalized lymphadenopathy, which was mistakenly treated as tubercular lymphadenopathy. A diagnosis of late bacteriological relapse of lepromatous leprosy presenting with prominent lymphadenopathy and ENL was made after relevant investigations. The patient was started on treatment with WHO MDT MB (daily dapsone and clofazimine and monthly rifampicin) and thalidomide (200 mg/day). Nerve pain regressed within 2 weeks of therapy. The lymph nodal swelling regressed within 3 months of starting treatment. PMID- 19293509 TI - Calcinosis cutis on the face. PMID- 19293510 TI - Plagiarism, copyright violation, duplication, fabrication, false statements, breach of research and publication ethics. PMID- 19293511 TI - Dexamethasone-cyclophosphamide pulse therapy in pemphigus. PMID- 19293512 TI - Halobetasol versus clobetasol: a study of potency. PMID- 19293513 TI - Effect of warfarin on chronic idiopathic urticaria. PMID- 19293514 TI - Our experience of the use of thalidomide in the steroid-dependent severe erythema nodosum leprosum. PMID- 19293515 TI - Leprosy in the era of integration. PMID- 19293516 TI - Enhancing digital images using unsharp-mask. PMID- 19293517 TI - Eccrine angiomatous hamartoma in an adult. PMID- 19293518 TI - Doctor I am on fire: eosinophilic folliculitis in HIV negative. PMID- 19293519 TI - Verruca plana following eyebrow threading. PMID- 19293520 TI - Blepharochalasis. PMID- 19293521 TI - Tumor d'emblee responding to methotrexate and prednisolone. PMID- 19293522 TI - Rupioid syphilis in an HIV patient. PMID- 19293523 TI - Becker's nevus with ipsilateral breast hypoplasia. PMID- 19293526 TI - Pap smear. PMID- 19293527 TI - Asymptomatic papular lesions on the trunk. PMID- 19293528 TI - Chylomicronemia syndrome. PMID- 19293529 TI - Impact of psoriasis on quality of life in Iran. AB - BACKGROUND: The Psoriasis Disability Index (PDI) questionnaire is a widely used instrument to measure psychological morbidity in plaque-type psoriasis patients. AIMS: This study aimed to validate the Persian version of the PDI and to evaluate the impact of psoriasis on quality of life (QOL). METHODS: The English language version of the PDI was translated into Persian (Iranian official language) and was used in this study. The questionnaire was administered to a consecutive sample of 125 chronic plaque-type psoriasis patients and statistical analysis was performed to evaluate the impact of psoriasis on QOL. The other health-related QOL assessment tool included the Persian version of the Dermatology Life Quality Index (DLQI). RESULTS: Overall, 125 patients who had received the PDI and DLQI completed all the questions. Reliability analysis showed a satisfactory result (Cronbach's a coefficient=0.92 and 0.79 for PDI and DLQI, respectively). There was a strong statistical correlation between mean PDI and DLQI scores, with mean Psoriasis Area and Severity Index (PASI) (P=0.005 and 0.02). Also, a significant correlation coefficient existed between DLQI and PDI (r=0.94). The higher the PASI index, the higher the PDI and DLQI scores, which indicated greater impact on QOL. In the patients with lesions on visible exposed skin areas, the correlation was statistically significant (P=0.002 and 0.01). CONCLUSION: The Persian PDI is an acceptable, reliable and valid measure of psychological distress, with more suitable content validity than DLQI for assessment of impact of psoriasis on QOL among psoriasis patients. Data provided may improve the physicians' awareness of the importance of the patients' QOL. PMID- 19293530 TI - Servo-controlled hind-limb electrical stimulation for short-term arterial pressure control. AB - BACKGROUND: Autonomic neural intervention is a promising tool for modulating the circulatory system thereby treating some cardiovascular diseases. METHODS AND RESULTS: In 8 pentobarbital-anesthetized cats, it was examined whether the arterial pressure (AP) could be controlled by acupuncture-like hind-limb electrical stimulation (HES). With a 0.5-ms pulse width, HES monotonically reduced AP as the stimulus current increased from 1 to 5 mA, suggesting that the stimulus current could be a primary control variable. In contrast, the depressor effect of HES showed a nadir approximately 10 Hz in the frequency range between 1 and 100 Hz. Dynamic characteristics of the AP response to HES approximated a second-order low-pass filter with dead time (gain: -10.2 +/- 1.6 mmHg/mA, natural frequency: 0.040 +/- 0.004 Hz, damping ratio 1.80 +/- 0.24, dead time: 1.38 +/- 0.13 s, mean +/- SE). Based on these dynamic characteristics, a servo-controlled HES system was developed. When a target AP value was set at 20 mmHg below the baseline AP, the time required for the AP response to reach 90% of the target level was 38 +/- 10 s. The steady-state error between the measured and target AP values was 1.3 +/- 0.1 mmHg. CONCLUSIONS: Autonomic neural intervention by acupuncture-like HES might provide an additional modality to quantitatively control the circulatory system. PMID- 19293531 TI - Relation between abnormalities in circadian blood pressure rhythm and target organ damage in normotensives. AB - BACKGROUND: To determine the individual effect of abnormalities in blood pressure (BP) circadian rhythm (non-dipping status (NDS), increased morning BP (MBP) or increased MBP surge (MBPS)) on target organ damage (TOD) and which of these is more closely related to TOD in normotensives. METHODS AND RESULTS: The 24-h ambulatory BP monitoring (ABPM) and echocardiography were performed and urinary albumin excretion (UAE) was measured in 47 dipper (28 women, mean age 45.8 +/- 9.3) and 32 non-dipper (25 women, mean age 49.1 +/- 8.3 years) normotensive subjects. The left ventricular mass index (LVMI) was higher in non-dipper group (103.8 +/- 24.1 vs 91.6 +/- 23.5 g/m2, P=0.03). UAE in non-dipper group was higher, but the difference between the two was not statistically significant (18.9 [10.3, 28.9] vs 14.1 [7.5, 23.8], P=0.11). In multivariate analysis, both LVMI and UAE were affected by NDS and MBP independent of other confounding variables (for LVMI; Coefficient =0.27, P=0.01 and Coefficient =0.37, P=0.001, respectively, and for UAE; Coefficient =0.27, P=0.02 and Coefficient =0.28, P=0.01, respectively). CONCLUSIONS: It may be postulated that increased night and MBP are the factors that cause TOD, and it seems reasonable to attempt to restore normal diurnal rhythm of the BP even in normotensive subjects. PMID- 19293532 TI - Plasma tetrahydrobiopterin/dihydrobiopterin ratio: a possible marker of endothelial dysfunction. AB - BACKGROUND: Although endothelium-dependent vasodilatation has been used as a marker of endothelial dysfunction (ED), there have been no reliable plasma markers for ED. Oxidative stress, which is a major determinant of ED, oxidizes tetrahydrobiopterin (BH4), an essential cofactor of endothelial type nitric oxide synthase (eNOS), and resulted in the relative deficiency of BH4. METHODS AND RESULTS: In 163 patients with cardiovascular disorders, the plasma levels of BH4 and 7, 8-dihydrobiopterin (BH2) by high performance liquid chromatography were measured and compared with the flow-mediated (FMD) vasodilatory response of the brachial artery, which was measured by ultrasonography. The effects of atorvastatin on plasma pteridine levels and FMD were examined in patients with multiple coronary risk factors. There was a positive relationship between FMD and plasma BH4 levels and a negative relationship between FMD and plasma BH2 levels. Subsequently, a strong positive relationship between FMD and the BH4/BH2 ratio (r=0.585, P<0.0001) was found. Although we did not find any significant relationship between pteridine levels and individual traditional risk factors, the BH4/BH2 ratio in patients with more than 2 risk factors showed significant reductions compared with that in those without risk factors. Statin treatment improved FMD in association with an increase in the plasma BH4/BH2 ratio. CONCLUSIONS: Plasma pteridine levels were associated with endothelial dysfunction in cardiovascular disorders. PMID- 19293533 TI - Coexistent true aortic aneurysm as a cause of acute aortic dissection. AB - BACKGROUND: Aortic aneurysms are found in 5-20% of patients with acute aortic dissection (AAD). Coexisting aortic aneurysms might potentially influence the incidence of AAD. The purpose of this study was to elucidate the role of coexistent aortic aneurysms in AAD. METHODS AND RESULTS: A total of 140 patients with AAD were enrolled in the present study. Clinical characteristics of the patients were evaluated in relation to the locations of aortic segments affected by the dissection as well as of the coexistent aortic aneurysm. Among the 140 study patients, 34 (24%) had true aortic aneurysms. Patients with coexistent aortic aneurysm were significantly older than those without (72 +/- 11 years vs 65 +/- 14 years, P=0.012) and had higher incidence of thrombosed false lumen (62% vs 38%, P=0.017), and coronary artery disease (26% vs 8%, P=0.006). Twenty-two of these 34 (65%) patients had a thoracic aortic aneurysm (TAA), and this frequency of TAA was much higher than that observed in the general population. Furthermore, among all patients with AAD, 12 patients (9%) might be associated with development of AAD. CONCLUSIONS: The current study showed that nearly one-quarter of AAD patients had coexisting true aortic aneurysms, and suggests that TAA are likely to be associated with development of AAD. PMID- 19293534 TI - Magnetic separation of human podocalyxin-like protein 1 (hPCLP1)-positive cells from peripheral blood and umbilical cord blood using anti-hPCLP1 monoclonal antibody and protein A expressed on bacterial magnetic particles. AB - Hemangioblasts are common progenitors of hematopoietic and angiogenic cells, which have been demonstrated in the mouse to possess a unique cell surface marker, podocalyxin-like protein 1 (PCLP1) (Hara, T. et al., Immunity, 11: 567 578. 1999). In this study, we prepared a novel monoclonal antibody against human PCLP1 (hPCLP1) and attempted to isolate human hematopoietic progenitor cells from umbilical cord blood and peripheral blood using nano-sized bacterial magnetic particles (BacMPs) coupled with the anti-hPCLP1 antibody. Flow cytometric analysis demonstrated that the purity of separated hPCLP1-positive cells from peripheral blood was approximately 95% whereas peripheral blood mononuclear cells contained only 0.1% PCLP1+ cells. Umbilical cord blood was demonstrated to be a better source for PCLP1+ cells than peripheral blood. These results suggest that the separation of human PCLP1+ cells using BacMPs with anti-hPCLP1 were extremely effective and may be useful as a means to prepare human hematopoietic progenitor cells. PMID- 19293535 TI - Novel function of transcription factor ATF5: blockade of p53-dependent apoptosis induced by ionizing irradiation. AB - PURPOSE: To find a new molecule that affects p53-dependent radiosensitivity. METHODS AND MATERIALS: A mouse sarcoma cell line, QRsP(p53+/+), was used. From this cell line, we established a radiosensitive clone and a radioresistant one. Colony assay, p53 gene transfer, a luciferase assay for p53 and p21, animal transplantation experiment, and DNA array analyses were performed. RESULTS: Microarray showed marked reduction of a transcription factor, ATF5, both in vitro and in vivo for the radiosensitive clone. Interestingly, flow cytometric analysis demonstrated marked apoptosis for the radiosensitive clone by p53 cloned adenovirus infection. Luciferase reporter assay revealed that ATF5 suppressed the transactivational activity of p53 and p63. By ATF5 gene transfer, the radiosensitive clone regained resistance to both ionizing-radiation and Ad-p53 infection-induced cell death. Surprisingly, time-lapse cell migration observation revealed greater cell motility for ATF5-transfected radiosensitive clone. CONCLUSIONS: It seems likely that ATF5 is a potent repressor of p53 and elevated expression of ATF5 in a tumor may relate to enhanced malignant phenotypes, such as radioresistance or greater cell motility. PMID- 19293536 TI - Brain natriuretic peptide (BNP) level is closely related to the extent of left ventricular sympathetic overactivity in chronic ischemic heart failure. AB - OBJECTIVE: Both brain natriuretic peptide (BNP) and cardiac sympathetic activity are useful surrogate markers of congestive heart failure. BNP is known to be secreted in response to sympathetic tone. This study examined the relationship between the cardiac sympathetic system and BNP. PATIENTS AND METHODS: Sixty patients with chronic ischemic heart failure (mean age,72 years-old; 46 males and 14 females) who had undergone cardiac catheterization and were classified as NYHA II underwent resting (99m)Tc-sestamibi quantitative gated imaging (MIBI) and (123)I-metaiodobenzylguanidine imaging (MIBG). MIBI was used to obtain left ventricular (LV) dimension. MIBG was used to obtain the washout rate and the H/M ratios as well as the extent of LV washout rate abnormality (RSNA), which was defined as the number of regions with a regional washout rate of more than mean+2SD of 15 normal subjects on a two-dimensional polar map divided into 20 regions. Blood samples were obtained to measure neurohormones such as BNP, renin activity, noradrenaline, and angiotensin II. RESULTS: Simple linear regression analysis showed that BNP had significant correlations to age, LVEF, LV end diastolic volume, LV end systolic volume, RSNA, global washout rate, myocardial ischemia, and LV end diastolic pressure. Among them, multiple linear regression analysis showed that only RSNA (partial regression coefficient =0.618, p<0.002) had a significant positive correlation with BNP. CONCLUSION: This study suggested that the extent of cardiac sympathetic overactivity in the LV could enhance BNP release. PMID- 19293537 TI - Risk assessment by post-challenge plasma glucose, insulin response ratio, and other indices of insulin resistance and/or secretion for predicting the development of type 2 diabetes. AB - OBJECTIVE: To find useful indices to identify subjects at high risk for developing diabetes. METHODS: We retrospectively reviewed 75-g oral glucose tolerance test (OGTT) surveys conducted during 1980 to 2001 in a Japanese community. Using personal charts of the surveys, 230 non-diabetics were followed for progression to type 2 diabetes. The usefulness of HbA(1C), fasting (Glucose(0)) and 1-hour (Glucose(60)) glucose levels during OGTT, and indices for insulin resistance and/or secretion to identify high risk subjects for diabetes were then analyzed. MATERIALS: Data described in personal charts of the OGTT program for residents of Nishikawa Town, Niigata Prefecture, Japan. RESULTS: During the 4.3+/-2.7 years of follow-up, 52 subjects progressed to type 2 diabetes. Assessing glucose and insulin levels during OGTT, Glucose(0), Glucose(60), "Insulin Response Ratio" (IRR(30), Insulin at 30 minutes / Fasting insulin; IRR(60), insulin at 60 minutes / fasting insulin), and insulin secretion / insulin resistance index (ISIRI(30)) were significantly associated with progression to type 2 diabetes even after the result for OGTT, body mass index, and familial history of diabetes were adjusted. These were also able to identify still higher risk subjects for type 2 diabetes from those with impaired glucose tolerance (IGT) although other indices for insulin resistance or secretion and hemoglobin A(1C) were less contributable for this purpose. CONCLUSION: A combination of Glucose(0) and Glucose(60) can most cost effectively identify high risk subjects for type 2 diabetes from IGT. IRR(30), IRR(60) and ISIRI(30) can also be used for such identification. However, further studies are needed to clarify whether these indices are superior to Glucose(0) and Glucose(60). PMID- 19293538 TI - A cross-sectional survey of chronic kidney disease and diabetic kidney disease in Japanese type 2 diabetic patients at four urban diabetes clinics. AB - OBJECTIVE: Management of chronic kidney disease (CKD) is a critical issue in public health in attempt to prevent kidney failure and dialysis dependency. Since 1998, diabetes mellitus has been the leading cause of dialysis dependency in Japan. Previous reports demonstrated that the prevalence of CKD in diabetic patients was high; however albuminuria was not always present. This cross sectional survey was performed 1) to indicate the prevalence of CKD and co-morbid illness in diabetic patients seen at diabetic clinics, and 2) to demonstrate the relationship between estimated glomerular filtration rate (eGFR) and urine albumin to creatinine ratio (ACR). PATIENTS AND METHODS: A total of 288 consecutive adult type 2 diabetic patients seen at four diabetic clinics in the Tokyo Metropolitan Area were enrolled in November 2007. We excluded patients with kidney failure. Estimated GFR was calculated by the MDRD Study equation with the Japanese coefficient. RESULTS: Patients had a mean age (+/- SD), 61 (+/- 12); male, 58%; mean BMI, 25.2 kg/m(2) (+/- 5.2); and mean HbA(1c), 7.1% (+/- 1.3). The prevalence of CKD stage 3 was 38% (109/288) with 64% (70/109) of them being normoalbuminuric. Co-morbid illnesses, including hypertension (p<0.001) and old stroke (p=0.02), were significantly higher in CKD stage 3 patients. CONCLUSION: Our patients were relatively young and obese, reflecting urban clinical settings. The prevalence of CKD stage 3 patients was high. Clinicians need to check both eGFR and ACR to avoid underdiagnosis of CKD and diabetic kidney disease. PMID- 19293539 TI - Sevelamer decreases serum uric acid concentration through adsorption of uric acid in maintenance hemodialysis patients. AB - BACKGROUND: Sevelamer, a nonabsorbed hydrogel that binds phosphate, is reported to reduce the serum urate concentration in maintenance hemodialysis patients, however the urate-lowering mechanism remains obscure. In this study we verify the urate-lowering effect of sevelamer in Japan in which the hemodialysis environment is different from that of western countries, and we also clarify the urate lowering mechanism of sevelamer. METHODS: A total of 127 Japanese patients undergoing maintenance hemodialysis were investigated. These patients consisted of 93 males and 34 females, and their mean age was 58.4+/-12.4 years (range, 25 88 years). The mean duration of hemodialysis was 8.7+/-6.1 years (range, 0.5-27.5 years). Sevelamer was added to each patient's former prescription for the treatment of hyperphosphatemia, and the changes in laboratory data before and after administration of sevelamer were compared. In order to clarify the mechanism of urate-lowering effect by sevelamer, a urate adsorption experiment was carried out in vitro. RESULTS: Sevelamer significantly decreased serum phosphate value three and six months after administration. Sevelamer showed a significant reduction in serum urate values in maintenance hemodialysis patients with hyperuricemia, but not in patients with normouricemia. The change rate of serum urate correlated with the change rate of serum phosphate and the change rate of serum calcium x phosphate product, but did not correlate with that of serum calcium. Sevelamer hydrochloride adsorbed urate in vitro. CONCLUSION: Sevelamer decreases serum urate possibly by adsorbing urate in hemodialysis patients. PMID- 19293540 TI - Circulating hepcidin in the crossroads of anemia and inflammation associated with rheumatoid arthritis. AB - OBJECTIVE: To investigate the role of circulating hepcidin, which is a homeostatic regulator of iron metabolism and a mediator of inflammation, in anemia associated with rheumatoid arthritis. METHODS: Forty patients with rheumatoid arthritis (19 with anemia and 21 without anemia), 12 patients with iron deficiency anemia and 14 healthy adults were studied. Serum hepcidin levels were analyzed with hepcidin prohormone solid phase enzyme-linked immunosorbent assay. RESULTS: The mean serum hepcidin levels were significantly higher in patients with rheumatoid arthritis with anemia compared to healthy adults and those with iron deficiency anemia. The active rheumatoid arthritis group had significantly higher mean serum hepcidin levels than the inactive rheumatoid arthritis group. In the rheumatoid arthritis group, serum hepcidin levels were positively correlated with disease activity, but inversely correlated with hemoglobin levels. The serum hepcidin level was found to be a significant predictor for hemoglobin level. CONCLUSION: Serum hepcidin levels are closely associated with disease activity in rheumatoid arthritis patients and might play a role in the pathobiology of chronic disease anemia associated with rheumatoid arthritis. PMID- 19293541 TI - Characteristics of Japanese patients with complex sleep apnea syndrome: a retrospective comparison with obstructive sleep apnea syndrome. AB - OBJECTIVE: The prevalence of complex sleep apnea syndrome (CompSAS) among Asian patients with obstructive sleep apnea syndrome (OSAS) has not yet been reported. Distinguishing CompSAS from pure OSAS is difficult using only diagnostic polysomnography (PSG). We examined the prevalence of CompSAS in Japanese patients with OSAS and the possibility to distinguish CompSAS from pure OSAS by analyzing the severity of respiratory events based on either sleep body position or sleep stage using a diagnostic PSG. PATIENTS AND METHODS: A retrospective chart review of 297 consecutive Japanese patients who were 15 years of age or older with a primary diagnosis of OSAS who were referred for CPAP titration (AHI> or =20 events/hr). RESULTS: Seventeen patients (5.7%) out of the 297 patients who had an obstructive apnea hypopnea index (AHI) of 20 or higher showed adverse increases in central apnea index (CAI) by the treatment with CPAP whereas obstructive apnea index (OAI) and mixed apnea index (MAI) were significantly decreased. In the results, the AHI on the PSG for CPAP titration reached only approximately half of the values on the diagnostic PSG. In these CompSAS patients, both the total CAI and the CAI in the supine position during NREM sleep on the diagnostic PSG were significantly higher than those in the OSAS group. The sleep body position did not so strongly affect the AHI, OAI and MAI in the CompSAS group. Multiple, stepwise, and logistic regression analyses revealed that the CAI in the supine position during NREM (p=0.026) was a significant variable to distinguish CompSAS from OSAS statistically although the variables were within the normal range. CONCLUSION: The prevalence of CompSAS in Japanese OSAS patients may be lower when compared with Caucasian patients. The increase of CAI in the supine position during NREM sleep on diagnostic PSG may be a characteristic feature in CompSAS. PMID- 19293542 TI - Pulmonary embolism due to internal jugular vein thrombosis without an indwelling catheter. AB - A 65-year-old man who had undergone retropubic prostatectomy for prostate adenocarcinoma presented with sudden dyspnea and chest pain. Contrast-enhanced multi-slice computed tomography (MSCT) revealed thrombi in the left internal jugular vein (IJV) and in branches of the right pulmonary artery. Ultrasonography showed that the thrombus which occluded the left IJV was hypoechoic and mobile. After beginning anticoagulant therapy, he again presented with dyspnea and transient hypotension. MSCT and ultrasonography showed that the IJV thrombus had moved and caused a new embolism of the left pulmonary artery branch. This is a rare case of a patient who experienced non-catheter-related thrombosis of the IJV. PMID- 19293543 TI - Compound heterozygous mutation of aquaporin 2 gene in woman patient with congenital nephrogenic diabetes insipidus. AB - We performed mutational analyses of a woman patient with congenital nephrogenic diabetes insipidus referred to us during pregnancy. The diagnosis was made during the neonatal period, after which she was treated with spironolactone and hydrochlorothiazide. Our examination showed the patient to be apparently in good health without definite evidence of dehydration. Serum and urine osmolality were 220 mOsm/L and 50 mOsm/L, respectively, and the serum concentration of AVP was 2.7 pg/mL. Results of a water-deprivation test performed after delivery were compatible with nephrogenic diabetes insipidus. Mutational analyses showed that the patient was a compound heterozygote with point mutations at nucleotide position 298 (G to A; G100R) in exon 1 and nucleotide position 374 (C to T; T125M) in exon 2 of the aquaporin 2 gene, which have been previously described. PMID- 19293544 TI - Pituitary abscess with panhypopituitarism showing T1 signal hyperintensity of the marginal pituitary area: a non-invasive differential diagnosis of pituitary abscess and pituitary apoplexy. AB - A 53-year-old man was hospitalized with general fatigue, headache, dizziness and polyuria. The laboratory findings revealed anterior hypopituitarism and central diabetes insipidus. He also showed eye movement disorder and facial sensory impairment. These symptoms were treated successfully with conservative medical treatment. Concurrently, abnormal pituitary MR imaging findings were revealed. Pituitary abscess was primarily suspected on MR imaging findings, although it was difficult to differentiate pituitary apoplexy by MR imaging findings, alone. In this report, we propose a new diagnostic approach of pituitary abscess, using a combination of CT, MR imaging and clinical manifestations, without either pituitary surgery or pituitary biopsy. PMID- 19293545 TI - Acromegaly accompanied by Turner syndrome with 47,XXX/45,X/46,XX mosaicism. AB - A 33-year-old woman was hospitalized for examination of edematous laryngopharynx. She was acromegalic. A pituitary adenoma with elevated serum levels of growth hormone (GH) and insulin-like growth factor-I (IGF-I) was detected, indicating acromegaly caused by GH-secreting pituitary adenoma. Multiple pigmented nevi were also noted without overt short stature and cubitus valgus. Chromosome analysis revealed that she had contracted Turner syndrome with 47,XXX/45,X/46,XX mosaicism. Transsphenoidal resection of the tumor decreased serum GH and IGF-I levels, but the edema was not improved. Both premature ovarian failure and hypertension appeared after surgery. This case may indicate the important relationships between GH/IGF-I and Turner syndrome. PMID- 19293546 TI - Catheter-related bacteremia caused by Agrobacterium radiobacter in a hemodialysis patient. AB - Agrobacterium radiobacter, a Gram-negative bacillus, is recognized as an emerging opportunistic human pathogen that has a propensity to cause infections in patients with indwelling foreign devices. Here, we describe the first reported case of catheter-related bacteremia caused by A. radiobacter in a hemodialysis patient with a long-term tunneled-cuffed hemodialysis catheter. This case shows that A. radiobacter should be included in the list of pathogens that can cause catheter-related bacteremia in hemodialysis patients. PMID- 19293547 TI - Successful treatment with pneumonectomy for pulmonary Mycobacterium abscessus infection. AB - We report a case of pulmonary Mycobacterium abscessus (M. abscessus) infection with destructive growth in the entire right lung. The patient was 56-year-old woman who had had pulmonary tuberculosis at the age of 40 and had been diagnosed as having pulmonary Mycobacterium abscessus infection 4 years prior to admission at our hospital. Although various antibiotics were administered, persistent fever, hemoptysis and weight loss developed. After undergoing a right pneumonectomy, her clinical symptoms improved dramatically and sputum excretions of M. abscessus ceased. No relapse of the disease has been observed in the 2 years since surgery. Pneumonectomy was very effective for refractory M. abscessus infection that destroyed the right lung. PMID- 19293548 TI - Transient severe pancytopenia due to elevated tumor necrosis factor-alpha in overwhelming infection. AB - A 66-year-old woman complained of fever, sore throat, and neck pain due to pharyngitis and painful lymph node swelling. CBC revealed severe pancytopenia and markedly hypocellular marrow. The administration of antibiotics and granulocyte colony stimulating factor (G-CSF) successfully ameliorated the inflammatory lesions, and hematopoiesis recovered. Causes for pancytopnenia was unlikely to be virus infection or drugs, and aplastic anemia was also unlikely since only the plasma levels of tumor necrosis factor-alpha (TNF-alpha) was markedly elevated, erythropoietin (EPO) was slightly elevated, interferon-gamma (IFN-gamma) was normal, and flow cytometric analysis for paroxysmal nocturnal hemoglobinuria (PNH)-type cells was negative. These results suggested that the cause of impaired hematopoiesis in the present patient might have been due to elevated TNF-alpha in overwhelming infection, although the pathogen was not identified. PMID- 19293549 TI - Adult patient with Epstein-Barr virus (EBV)-associated lymphoproliferative disorder: chronic active EBV infection or de novo extranodal natural killer (NK)/T-cell lymphoma, nasal type? AB - Chronic active Epstein-Barr virus (EBV) infection, which is considered to be a childhood disease, often develops into natural killer (NK) or T-cell lymphoma after recurrent infectious mononucleosis (IM)-like symptoms. We describe a 56 year-old woman who developed NK-cell lymphoma after 9 months of recurrent IM-like symptoms. The patient had an unusual anti-EBV antibody profile. Increased levels of EBV genome were detected in the liver and peripheral blood. Several chemotherapy regimens were ineffective, and the patient died of progression of lymphoma. Certain subtypes of NK-cell lymphoma showing a long-lasting prodrome related to chronic EBV infection may exist. PMID- 19293550 TI - Intensive chemotherapy for a patient with primary cutaneous diffuse large B-cell lymphoma with Burkitt-like morphology. AB - We report a rare case of primary cutaneous diffuse large B-cell lymphoma (DLBCL) with Burkitt-like morphology. A 54-year-old man presented with multiple subcutaneous tumors. Pathological examination showed morphological features resembling Burkitt or Burkitt-like lymphoma (BL/BLL) with high MIB-1 positivity. Cytogenetic studies revealed no 8q24/c-myc translocation. After the diagnosis of Burkitt-like DLBCL, the patient was treated with CODOX-M chemotherapy (cyclophosphamide, doxorubicin, vincristine, cytarabine and methotrexate), which led to durable remission. The present case suggests that short-term, high intensity chemotherapy used for BL/BLL may be appropriate for primary cutaneous Burkitt-like DLBCL, as well as systemic lymphoma with Burkitt-like morphology. PMID- 19293551 TI - Acute disseminated encephalomyelitis developed after Mycoplasma pneumoniae infection complicating subclinical measles infection. AB - A 26-year-old man developed acute disseminated encephalomyelitis (ADEM) after Mycoplasma pneumoniae infection, and was admitted after developing disturbed consciousness. Magnetic resonance images revealed lesions in the midbrain, bilateral internal capsules, left corona radiata, white matter of the left occipital lobe, and thoracic spinal cord. He was diagnosed with subclinical measles infection since no anthema was observed despite the fact that his serum and cerebrospinal fluid samples were positive for measles IgM antibodies. ADEM following mixed infection with measles and M. pneumoniae is rare, and it is not clear whether an additional infection with measles influenced the onset of ADEM after M. pneumoniae infection. Symptoms did not improve with steroid or immunoglobulin treatment, but improvement in symptoms was observed after plasmapheresis. PMID- 19293552 TI - Brain abscess caused by pulmonary arteriovenous fistulas without Rendu-Osler Weber disease. AB - A 52-year-old man was admitted with a brain abscess in the left basal ganglia. He had a paradoxical brain embolic mechanism owing to a pulmonary arteriovenous fistula (PAVF) and was diagnosed as having a right-to-left shunt by transesophageal echocardiography (TEE) and transcranial color Doppler (TCD) with saline contrast medium. We determined that the brain abscess was caused by the PAVFs without Rendu-Osler-Weber disease. TEE and TCD with saline contrast medium were very useful for identifying the presence of the right-to-left shunt that caused the brain abscess. PMID- 19293553 TI - Histopathology of bone marrow "clot" section of a mantle cell lymphoma. PMID- 19293554 TI - Hypothalamic lymphoma with symptoms mimicking pituitary apoplexy. PMID- 19293555 TI - A truly unusual overgrowth syndrome: an alternative diagnosis to Klippel Trenaunay-Weber syndrome. PMID- 19293557 TI - Adding essential amino acids at a low concentration improves the development of in vitro fertilized porcine embryos. AB - The aims of this study were to investigate improvements to the pig preimplantation embryo culture system using in vitro produced embryos. For experiment 1, the optimum time to change the medium from NCSU23 containing 0.6 mM glucose, 0.2 mM pyruvate, 5.7 mM lactate and nonessential amino acids to NCSU23 containing 5.6 mM glucose and both essential and nonessential amino acids was examined. There were no statistically significant differences in blastocyst rates or cell number when the medium was changed at 48, 72 or 96 h, although there was a consistent trend for the 96 h treatment to produce fewer blastocysts with fewer cells. For experiment 2, the addition of essential amino acids at either a 1:50 or a 1:100 dilution of the purchased stock solution for day 1 to 6 or for days 3 to 6 only was investigated. Adding essential amino acids at a 1:50 dilution for day 3 to 6 significantly reduced the blastocyst rate and adding them at a 1:50 dilution from day 1 to 6 significantly reduced both the blastocyst rate and blastocyst cell number compared to when it was added at a 1:100 dilution. Embryos were produced by IVF, cultured for 6 days and good quality blastocysts were transferred into 6 synchronized pseudopregnant recipients (24 to 35 blastocysts per recipient) resulting in 4 pregnancies and 21 live birth piglets. These results show that adding essential amino acids at a 1:100 dilution provided the best culture conditions and the blastocysts produced were able to attain full term development after transfer. PMID- 19293558 TI - Cloning and characterization of cat POU5F1 and NANOG for identification of embryonic stem-like cells. AB - POU5F1 and NANOG play important roles in the maintenance of embryonic stem cell pluripotency. Recently, we isolated cat embryonic stem (ES)-like cells from cat blastocysts generated in vivo. In an effort to identify genetic markers for the characterization of cat ES-like cells, we have determined the coding sequences (CDSs) of cat POU5F1 (cPOU5F1) and NANOG (cNANOG). The sequence identities of cPOU5F1 with orthologous genes of the human and mouse were 92 and 82%, respectively, at the nucleotide level and 94 and 83%, respectively, at the amino acid level. We identified POU-specific and POU homeodomain sequences in the CDS of cPOU5F1. The sequence identities of cNANOG with its human and mouse orthologs were 69 and 68%, respectively, at the nucleotide level and 69 and 58%, respectively, at the amino acid level. We identified a homeodomain, SMAD4 domain and tryptophan repeat domain (W/QXXXX) in the CDS of cNANOG. We examined the expression of cPOU5F1 and cNANOG mRNA in ES-like cells and fibroblast feeder cells by RT-PCR. Transcripts of cPOU5F1 and cNANOG were detected at a high level in ES-like cells. However, these two genes were undetectable in cat fibroblast feeder cells and 6 adult tissues. We also examined ES-like cells by immunocytochemistry and demonstrated that cPOU5F1 and cNANOG are present at high levels in cat ES-like cells and are undetectable in cat fibroblast feeder cells. These results confirmed that cat ES-like cells can be successfully isolated from in vivo-produced blastocysts and that the expression of cPOU5F1 and cNANOG can be used as a biomarker for characterization of cat ES-like cells. PMID- 19293559 TI - Difference in sensitivity to culture condition between in vitro fertilized and somatic cell nuclear transfer embryos in pigs. AB - We evaluated the developmental competence of somatic cell nuclear transfer (SCNT) embryos using in vitro embryo culture systems. Embryos were cultured in NCSU-23, NCSU-23 supplemented with essential and non-essential amino acids (NCSU-23aa) or modified PZM-5 supplemented with BSA instead of PVA (mPZM-5). The rates of blastocyst formation were significantly higher in the mPZM-5 group than in the other groups, regardless of the method of embryo production (38.0 vs. 25.3 or 29.1% for IVF, 18.2 vs. 8.7 or 9.4% for SCNT, respectively). The mean cell numbers of IVF and SCNT blastocysts were also significantly higher in mPZM-5 than in the other groups (62.0 vs. 42.3 or 43.0 for IVF, 46.5 vs. 29.4 or 31.3 for SCNT, respectively). Next, the embryos were cultured in mPZM-5 from days 0 to 4 and then in mPZM-5 (P/P), NCSU-23 (P/N) or NCSU-23aa (P/Naa) until day 6. The rates of blastocyst formation were similar among the 3 two-step culture systems in both embryo groups (36.2, 34.2, and 33.6% for IVF, 20.8, 14.1, and 17.2% for SCNT, respectively). The mean cell number in the IVF and SCNT blastocysts was significantly lower in P/N than in P/P and P/Naa (46.5 vs. 63.5 and 68.7 for IVF, 29.3 vs. 45.5 and 39.7 for SCNT, respectively). Next, we examined the effect of media on apoptosis in IVF and SCNT blastocysts. The apoptosis indices in the blastocysts derived from either NCSU-23 or mPZM-5 were analyzed by TUNEL assay. The apoptosis index of the SCNT blastocysts was significantly lower in mPZM-5 than in NCSU-23 (8.8 vs. 13.6%), whereas no such difference was observed between groups in the IVF embryos (5.1 vs. 4.4%). These data suggested that SCNT embryos were more easily affected by culture environment compared with IVF embryos, offering the possibility to further enhance the developmental competence of SCNT embryos by developing more appropriate culture conditions in pigs. PMID- 19293560 TI - Spontaneous endometrial hyperplasia in the uteri of IL-2 receptor beta-chain transgenic mice. AB - We found frequent and spontaneous proliferation of glandular epithelium and dilated cysts in the uteri of interleukin-2 receptor (IL2R) beta-chain transgenic (Tg2Rbeta) mice. The aim of this study was to examine the involvement of IL2R beta-chain in the pathogenesis of endometrial hyperplasia (EH). Mouse uteri and serum were collected from Tg2Rbeta and normal littermates (NL), which were classified into three groups according to age. The incidence of EH increased in an age-dependent manner in both types of mice. However, in old age, Tg2Rbeta mice showed more serious phenotypes of EH than NL. Immunohistochemical analysis revealed specific localization of IL2R beta-chain in the glandular epithelial cells, with a correlation to the degree of EH, not only in the Tg2Rbeta uteri but also in the NL uteri. Immunoreactions of CD3 and CD25 were detected in the uteri of Tg2Rbeta but were weak in the uteri of NL, and CD25-positive cells were distributed in the endometrial stroma and myometrium in the Tg2Rbeta mice. These findings suggest that the IL2R beta-chain induces growth potential for glandular epithelial cells and an immune-privileged condition mediated by CD25+regulatory-T cells. PMID- 19293561 TI - Calyculin A-sensitive protein phosphatases are involved in maintenance of progressive movement in mouse spermatozoa in vitro by suppression of autophosphorylation of protein kinase A. AB - Protein serine/threonine phosphorylation in mammalian sperm flagella has been considered to play important roles in regulation of motility. Protein phosphorylation state reflects balance of enzymatic activities between protein phosphatases and protein kinases [predominantly protein kinase A (PKA)]. The aims of this study were to disclose roles of protein phosphatases in the regulation of sperm motility and to provide evidence for suppression of PKA full activation by protein phosphatases in sperm flagella. Mouse epididymal spermatozoa were incubated with a cell-permeable protein phosphatase 1 (PP1)/protein phosphatase 2A (PP2A) inhibitor (calyculin A: 25-125 nM) at 37.5 C. After incubation, they were used for immunodetection of phosphorylated proteins, PKA and PP1 gamma2, assessment for motility and co-immunoprecipitation of PP1gamma2 with PKA. Incubation with calyculin A enhanced the phosphorylation states of several proteins (>250 kDa, 170 kDa, 155 kDa, 140 kDa and 42 kDa for serine/threonine phosphorylation and 70 kDa for tyrosine phosphorylation) and PKA catalytic subunits [at the autophosphorylation residue (Thr-197) for its full enzymatic activation] in the flagella. Coincidently, this incubation induced changes of sperm flagellar movement from the progressive type to the hyperactivation-like type. Indirect immunofluorescence and co-immunoprecipitation showed that PKA was co-localized with PP1 gamma2 in the principal pieces of sperm flagella. These findings suggest that calyculin A-sensitive protein phosphatases (PP1/PP2A) suppress full activation of PKA as well as enhancement of the phosphorylation states of other flagellar proteins in sperm flagella in order to prevent precocious changes of flagellar movement from the progressive type to hyperactivation. PMID- 19293562 TI - Local and systemic effects of embryos on uterine tissues during early pregnancy in pigs. AB - In the pig, the periimplantation period is critical for successful establishment of pregnancy. We studied the influence of embryos on oxytocin (OT) and progesterone (P(4)) regulated endometrial and myometrial secretion of 1) luteotrophic prostaglandin E(2) (PGE(2)) and 2) luteolytic prostaglandin F(2alpha) and its metabolite (PGFM) on days 12-14 of pregnancy in pigs. We used unilaterally pregnant pigs created by a surgical procedure in which one uterine horn remained intact and the second horn was cut transversely so that part of the horn was detached from the uterine body. The animals were divided into two groups, inseminated gilts (days 12-14 of pregnancy, n=6) and uninseminated cyclic gilts, which were used as controls (days 12-14 of estrous cycle, n=5). Embryos developed only in the patent part of the uterus and not in the occluded horn. The abundance of OTR mRNA was increased in the endometrium and decreased in the myometrium of the gravid uterine horn in the pregnant pigs compared with the non gravid uterine horn or either uterine horn in the cyclic pigs, indicative of a local effect of the conceptus. The presence of embryos in the uterine horn during the periimplantation period determines endometrial metabolism of PGF(2alpha) and the local response of the endometrium to OT and P(4). OT stimulates PGF(2alpha) secretion and PGFM accumulation in endometrial cultures only from the non-gravid uterine horn and controls PGE(2) secretion from the endometrium and myometrium in the pregnant gilts. The results indicate a more systemic affect of pregnancy on the uterine response to OT and a possibly the local effect of the conceptus in promoting progesterone's inhibition of OT-stimulated PGE(2) secretion and uterine metabolism of PGF(2alpha). PMID- 19293563 TI - Tetraspanin CD9 in bovine oocytes and its role in fertilization. AB - This study was conducted in bovine to investigate whether CD9 (a member of the tetraspanin superfamily of proteins) is present on oocytes and whether it functions in sperm-oocyte binding and fusion. First, the presence of CD9 in bovine matured oocytes was examined by immunofluorescence with the anti-CD9 monoclonal antibody (mAb) and fluorescein isothiocyanate-conjugated goat anti mouse antibody, and the results showed that CD9 was expressed on the plasma membrane of matured oocytes. Sperm binding and fusion with oocytes was then examined by in vitro fertilization. When the zona pellucida-free matured oocytes were fertilized, both sperm binding to ooplasma and sperm penetrating into oocytes were significantly (P<0.01) reduced in anti-CD9 antibody-treated oocytes (6.3 +/- 0.7 per oocyte and 41.6%, respectively) compared with untreated control oocytes (19.0 +/- 0.7 per oocyte and 81.3%, respectively), indicating that the anti-CD9 mAb potentially inhibits sperm-oocyte binding and fusion. These results demonstrated that the CD9 present on bovine matured oocytes is involved in sperm oocyte interaction during fertilization. PMID- 19293564 TI - Topical treatments with pimecrolimus, tacrolimus and medium- to high-potency corticosteroids, and risk of lymphoma. AB - BACKGROUND/AIMS: A potential risk of lymphoma associated with the use of topical calcineurin inhibitors is debated. We assessed the risk of lymphoma among patients treated with topical pimecrolimus, tacrolimus or corticosteroids. METHODS: We conducted a cohort study using health insurance claims data. Cohorts of initiators of topical pimecrolimus, tacrolimus and corticosteroids, along with cohorts of persons with untreated dermatitis and randomly sampled enrollees were identified from January 2002 to June 2006. Lymphomas were identified using insurance claims and adjudicated by medical records review. We adjusted for confounders by propensity score matching. RESULTS: Among 92,585 pimecrolimus initiators contributing 121,289 person-years of follow-up, we identified 26 lymphomas yielding an incidence of 21/100,000 person-years. This incidence of lymphoma was similar to that among tacrolimus users (rate ratio, RR = 1.16; 95% confidence interval, CI = 0.74-1.82) as well as corticosteroid users (RR = 1.15; 95% CI = 0.49-2.72). All three topical treatments were associated with an increased risk of lymphoma compared with the general population (RR(Pim) = 2.89; RR(Tac) = 2.82; RR(Cort) = 2.10) suggesting increased detection of preexisting lymphomas. CONCLUSION: This study did not find an increased risk of lymphoma among initiators of topical pimecrolimus relative to other topical agents during an average follow-up of 1.3 years. Longer-term studies may be needed. PMID- 19293565 TI - Urinary GADD45gamma expression is associated with progression of lgA nephropathy. AB - BACKGROUND: Growth arrest and DNA damage-45gamma (GADD-45gamma) is induced in response to environmental stresses and functions in the regulation of cell cycles. Previous findings by our group suggested that GADD45gamma contributed to renal tubular cell damage through induction of inflammatory and fibrogenic molecules. We examined the effects of urinary GADD45gamma expression on the decline in renal function with IgA nephropathy in the present study. METHODS: Patients (n = 62) were followed for a total of 710.3 +/- 287.5 days. The rate of renal function decline was assessed by the slopes of inverse serum creatinine and estimated glomerular filtration rate (GFR) plotted against time. Renal survival was calculated by Kaplan-Meier analysis and the primary endpoint was an increase in serum creatinine levels by 50% or more. RESULTS: Kidney function declined more rapidly in the GADD45gamma-positive group compared to the GADD45gamma-negative group. Kidney survival estimates at the end of the study period were 82.9% in the GADD45gamma-positive group and 100% in the negative group (p = 0.03). This difference remained significant in the group with GFR values <90 ml/min/1.73 m2 when adjusted to stratification factors. CONCLUSION: The results suggest that urinary GADD45gamma expression is associated with progression of renal disease. PMID- 19293566 TI - Flanking markers of cystatin c (CST3) gene do not show association with Alzheimer's disease. AB - BACKGROUND: In the present study we determined whether the cystatin c gene (CST3) is genetically associated with late-onset Alzheimer's disease (AD). METHODS: Two informative flanking single nucleotide polymorphisms (SNP), rs2424577 and rs3827143, of the CST3 gene and apolipoprotein E (APOE) gene were assessed in 568 Finnish AD patients and 688 cognitively healthy controls. Samples were genotyped with the TaqMan technique, and we conducted a single allele and genotypic distribution comparison as well as an estimation of haplotype frequencies between cases and controls. RESULTS: The APOE genotype distribution differed as expected between the AD cases and controls (p < 0.001). On the whole, any significant differences in AD risk were not found in single SNP and haplotype analyses for the CST3 gene between the whole study cohorts or in the stratified subgroups. Interestingly, AG-genotype carriers of rs3827143 showed a significant difference (p = 0.04) between cases and controls when compared to AA-genotype carriers, but this finding remained insignificant in the adjusted model. CONCLUSION: Although flanking SNP cover the whole gene transcript with strong linkage disequilibrium, our data show that the CST3 gene is not associated with AD risk in the Finnish population. PMID- 19293567 TI - Agreement in cognitive and clinical assessments in Alzheimer's disease. AB - AIMS: To estimate agreement among scores on three common assessments of cognitive function. METHOD: Baseline responses on the Alzheimer's Disease Assessment Scale Cognitive, Clinical Dementia Rating, and the Mini-Mental State Examination were obtained from two clinical trials (n = 138 and n = 351). A graphical method of examining agreement, the means-difference or Bland-Altman plot, was followed by Levene's test of the equality of variance corrected for multiple comparison within each sample. RESULTS: 70-78% of variability was shared by one factor, suggesting that all three instruments reflect cognitive impairment. However, agreement among tests was significantly worse for individuals with greater-than average, relative to individuals with less-than-average, cognitive impairment. CONCLUSIONS: Worse agreement between tests, as a function of increasing cognitive impairment, implies that interpretation of these tests and selection of coprimary cognitive impairment outcomes may depend on impairment level. PMID- 19293568 TI - The neuropsychological performance of nondemented Puerto Rican nonagenarians. AB - BACKGROUND/AIMS: While the oldest old are the fastest growing segment of the US population, normative neuropsychological data for nondemented oldest old Spanish speakers are nonexistent. This study sought to evaluate the neuropsychological performance of nondemented nonagenarians residing in Puerto Rico and to compare their results with those of a similar English-speaking sample from New York. METHODS: We studied 81 subjects who had a complete CERAD neuropsychological assessment in Spanish. We used multiple regression analysis to predict performance on the CERAD battery and ANCOVA to compare the Puerto Rico and New York samples. RESULTS: In 10 out of the 13 neuropsychological tests administered, education was a significant predictor of performance. There were significant differences between the Puerto Rico and New York groups only in the Trail Making Tests. CONCLUSIONS: In this Puerto Rican sample, education was the strongest predictor of neuropsychological performance, which is consistent with previous studies. When education level is properly accounted for, the performance of Puerto Rican nonagenarians in the CERAD battery does not differ from the performance of US English-speaking nonagenarians. PMID- 19293569 TI - Joint use of drugs and alcohol. AB - AIMS: To determine the relationships between the different patterns of drinking and drug use in the general population. DESIGN: The data from three nationwide surveys conducted in Finland in 1998, 2002 and 2004 were pooled into one dataset (n = 7,227). PARTICIPANTS: Population aged 15-69. MEASUREMENTS: Use of illicit drugs was analyzed in relation to annual alcohol consumption, heavy episodic drinking, frequenting of pubs and restaurants, and the misuse of prescribed drugs. Respondents were divided into five different groups according to their involvement in drug cultures, i.e. 'no contact with drug cultures', 'some contact with drug cultures', 'drug experimenters and ex-users', 'cannabis users' and 'multidrug users'. FINDINGS: The prevalence of drug use increased along with the annual alcohol consumption, and polysubstance use was most prevalent among heavy drinkers. Heavy episodic drinking and frequenting of pubs and restaurants increased in parallel with involvement in drug cultures. Misuse of prescribed drugs was most prevalent among multidrug users. CONCLUSIONS: Polysubstance use patterns reveal an interlacing of alcohol and drug cultures. However, standard measures employed in alcohol and drug research are insufficient to capture the phenomenon in detail, and more research and the development of new approaches are needed. PMID- 19293570 TI - Inherited and Sporadic Epimutations at the IGF2-H19 locus in Beckwith-Wiedemann syndrome and Wilms' tumor. AB - The parent-of-origin-dependent expression of IGF2 and H19 is controlled by the imprinting center 1 (IC1) consisting of a methylation-sensitive chromatin insulator. IC1 is normally methylated on the paternal chromosome and nonmethylated on the maternal chromosome. We found that 22 cases in a large cohort of patients affected by Beckwith-Wiedemann syndrome (BWS) had IC1 methylated on both parental chromosomes, resulting in biallelic activation of IGF2 and biallelic silencing of H19. These individuals had marked macrosomia and high incidence of Wilms' tumor. A subset of these patients had 1.4- to 1.8-kb deletions with hypermethylation of the remaining IC1 region and fully penetrant BWS phenotype when transmitted maternally. Another subset of individuals with IC1 hypermethylation had a similar clinical phenotype but no mutation in the local vicinity. All these cases were sporadic and in at least two families affected and unaffected members shared the same maternal IC1 allele but not the abnormal maternal epigenotype. Similarly, no IC1 deletion was detected in 10 nonsyndromic Wilms' tumors with IC1 hypermethylation. In conclusion, methylation defects at the IGF2-H19 locus can result from inherited mutations of the imprinting center and have high recurrence risk or arise independently from the sequence context and not transmitted to the progeny. PMID- 19293571 TI - Epigenetic regulation of growth: lessons from Silver-Russell syndrome. AB - Human growth is a complex process and requires the appropriate interaction of many members. Central members in the growth axes are regulated epigenetically and thereby reflect the profound significance of imprinting for correct mammalian ontogenesis. A prominent imprinting disorder, Silver-Russell syndrome (SRS), is a congenital disease characterized by intrauterine and postnatal growth retardation and other features. SRS represents the first human disorder with imprinting dis turbances affecting two different chromosomes: nearly one tenth of patients carry a maternal unipa-rental disomy of chromosome 7 and more than 38% show a hypomethylation in the imprinting control region 1 in 11p15. Interestingly, hypermethylation of the same region is associated with the overgrowth disease Beckwith-Wiedemann syndrome (BWS), thus SRS and BWS can be regarded as genetically (and clinically) opposite diseases. Because of the different imprinting regions involved, SRS is a suitable model to decipher the role of imprinting in growth and the functional interaction between imprinted genes in different genomic regions. PMID- 19293572 TI - Genetic imprinting: the paradigm of Prader-Willi and Angelman syndromes. AB - Imprinted genes are expressed from only one of the two parental alleles. A consequence of genomic imprinting is that viable embryos must receive two haploid genome complements from parents of opposite sex. The parental-specific expression is obtained through epigenetic modifications (DNA methylation, histone tail modifications) which alter the conformation of chromatin fiber and there-fore regulate the expression of the underlying genes. Deletions, duplication, mutations or alterations of imprinting of the only active allele, as well as uniparental disomy or loss of imprinting of the inactive allele lead to an unbalance (loss of function or gain of function) in the dosage of the gene product and may have phenotypic consequences. Two such examples in human pathology are rep-resented by the Prader-Willi and Angelman syndromes, whose phenotypes result from loss of paternal or maternal contribution of the 15 q11 q13 genomic region, respectively. Prader-Willi syndrome is characterized by pre- and postnatal hypotonia, feeding difficulties in early life and subsequent hyperphagia with obsessive/compulsive food searching, obesity, short stature, hypogonadism and acromicria. Psychomotor development is mildly affected and behavioral problems are more relevant. Patients with Angelman syndrome show a completely different phenotype characterized by severe mental retardation, absent speech, autistic-like behavior, severe epilepsy and postnatal microcephaly. PMID- 19293573 TI - Muscle involvement and IGF-1 signaling in genetic disorders: new therapeutic approaches. AB - In the last decade, dramatic progress has been made in elucidating the molecular defects underlying a number of muscle diseases. With the characterization of mutations responsible for muscle dysfunction in several inherited pathologies, and the identification of novel signaling pathways, subtle alterations in which can lead to significant defects in muscle metabolism, the field is poised to devise successful strategies for treatment of this debilitating and often fatal group of human ailments. Yet progress has been slow in therapeutic applications of our newly gained knowledge. The complexity of muscle types, the intimate relationship between structural integrity and mechanical function, and the sensitivity of skeletal muscle to metabolic perturbations have impeded rapid progress in successful clinical intervention. The relatively poor regenerative properties of striated muscle compound the devastating effects of muscle degeneration. Perhaps the most difficult hurdle is the sheer volume of tissue that must be treated to effect a significant improvement in quality of life. Recent studies on the role of insulin-like growth factor-1 in skeletal muscle growth and homeostasis have excited new interest in this important mediator of anabolic pathways and suggest promising new avenues for intervention in catabolic disease. In this review, we will discuss the potential therapeutic role of local insulin-like growth factor 1 in the treatment of muscle wasting associated with muscle diseases. PMID- 19293574 TI - Overgrowth syndromes: a classification. AB - Overgrowth syndromes are characterized by macrosomia, congenital anomalies, mental retardation and an increased risk of tumors. In this article we will analyze what we define 'classical' overgrowth. PMID- 19293575 TI - C-type natriuretic peptide and overgrowth. AB - Natriuretic peptides are a family of structurally related peptides with different distinct biological effects. C-type natriuretic peptide (CNP)-mediated signaling is important for endochondral ossification and intervenes in the control of chondrocyte maturation by regulating the balance between proliferation and terminal differentiation. CNP is encoded by the NPPC gene on human chromosome 2 for which, so far, no mutations have been described in humans. Recently, two independent articles reported the description of 3 patients with a similar clinical phenotype characterized by the presence of skeletal anomalies and overgrowth. In all 3 cases, the clinical picture was associated with the presence of a balanced translocation involving chromosome 2 and causing overexpression of the NPPC gene and an increased plasma concentration of its product, CNP. Transcriptional dysregulation of NPPC has been ascribed to the separation of the gene unit from the long-range regulatory element with a transcriptional silencing effect on its expression and CNP overproduction has been correlated to the skeletal overgrowth phenotype observed. PMID- 19293577 TI - Developmental abnormalities of the posterior pituitary gland. AB - While the molecular mechanisms of anterior pituitary development are now better understood than in the past, both in animals and in humans, little is known about the mechanisms regulating posterior pituitary development. The posterior pituitary gland is formed by the evagination of neural tissue from the floor of the third ventricle. It consists of the distal axons of the hypothalamic magnocellular neurones that shape the neurohypophysis. After its downward migration, it is encapsulated together with the ascending ectodermal cells of Rathke's pouch which form the anterior pituitary. By the end of the first trimester, this development is completed and vasopressin and oxytocin can be detected in neurohypophyseal tissue. Abnormal posterior pituitary migration such as the ectopic posterior pituitary lobe appearing at the level of median eminence or along the pituitary stalk have been reported in idiopathic GH deficiency or in subjects with HESX1, LHX4 and SOX3 gene mutations. Another intriguing feature of abnormal posterior pituitary development involves genetic forms of posterior pituitary neurodegeneration that have been reported in autosomal-dominant central diabetes insipidus and Wolfram disease. Defining the phenotype of the posterior pituitary gland can have significant clinical implications for management and counseling, as well as providing considerable insight into normal and abnormal mechanisms of posterior pituitary development in humans. PMID- 19293576 TI - Role of transcription factors in midline central nervous system and pituitary defects. AB - The anterior pituitary gland is a central regulator of growth, reproduction and homeostasis, and is the end-product of a carefully orchestrated pattern of expression of signalling molecules and transcription factors leading to the development of this complex organ secreting six hormones from five different cell types. Naturally-occurring and transgenic murine models have demonstrated a role for many of these molecules in the aetiology of combined pituitary hormone deficiency (CPHD). These include the transcription factors HESX1, PROP1, POU1F1, LHX3, LHX4, SOX2 and SOX3. The expression pattern of these transcription factors dictates the phenotype that results when the gene encod-ing the relevant transcription factor is mutated. The highly variable phenotype may consist of isolated hypopituitarism, or more complex disorders such as septo-optic dysplasia. Since mutations in any one transcription factor are uncommon, and since the overall incidence of mutations in known transcription factors is low in patients with CPHD, it is clear that many genes remain to be identified, and characterization of these will further elucidate the pathogenesis of these complex conditions, and also shed light on normal pituitary development. PMID- 19293578 TI - Hyperinsulinism in developmental syndromes. AB - Hyperinsulinism is a cause of recurrent and severe hypoglycaemia in the newborn and infancy period. Several developmental genetic syndromes are associated with hyperinsulinism. The underlying molecular mechanisms that lead to hyperinsulinaemic hypoglycaemia in most of these syndromes are unclear. Beckwith Wiedemann syndrome (BWS) is the most common syndrome associated with hyperinsulinism. The incidence of hyperinsulinism in children with BWS is about 50%. The hyperinsulinaemic hypoglycaemia can be transient, which, in the majority of infants, will be asymptomatic and resolve within the first few days of life. Rarely patients with BWS may require a pancreatectomy. Other overgrowth syndromes such as Soto's syndrome may overlap with BWS and present with hyperinsulinism. Patients with other rare syndromes such as Costello, Timothy and Kabuki syndromes can present with hyperinsulinaemic hypoglycaemia but the genetic mechanism(s) that leads to dysregulated insulin secretion in these syndromes is(are) still unclear. The congenital disorders of glycosylation (CDG) are a rapidly expanding group of metabolic syndromes with a wide symptomatology and severity. They all stem from deficient N-glycosylation of proteins. Hyper-insulinism has been described in congenital disorders of glycosylation, mostly in CDG-Ib but also as the leading symptom in a CDG-Ia patient. In summary, hyperinsulinism may be associated with a large number of developmental syndromes however the underlying molecular mechanisms that cause hyperinsulinism in these syndromes are still unknown. PMID- 19293579 TI - Developmental syndromes: growth hormone deficiency and treatment. AB - Developmental syndromes are characterized by numerous phenotypical signs and malformations. In most of them such as Turner, Noonan, Prader-Willi, Silver Russel, Williams, Kabuki, Leri-Weill syndrome and skeletal dysplasias, short stature is a common feature. Growth defect is very often related to a defect in cellular growth, but some unknown abnormality in GH action is possible. Recently, the greater availability of recombinant GH has expanded the interest towards GH secretion and therapy also in developmental syndromes. We recognize syndromes associated with GH deficiency (GHD), showing a developmental midline defect such as Pallister-Hall syndrome, septo-optic dysplasia, but many of these conditions do not have a convincing link with GHD. Moreover, some conditions, in particular the well-studied Turner syndrome, that do not have a real GHD, have proven to benefit from GH therapy at supra-physiological doses obtaining a higher final height than the expected one according to the natural history. This has expanded the indications for GH therapy. The aim of our paper is to review the literature on GH secretion, on the effects and costs-benefits of GH therapy in many dysmorphic syndromes, presenting some results of GH secretion and therapy obtained in our experience. PMID- 19293580 TI - Growth hormone-resistant syndromes: long-term follow-up. AB - Recombinant human IGF-I (rhIGF-I) has been approved as a therapeutic agent for short stature due to primary IGF-I deficiency following clinical trials that lasted more than 10 years. The first children treated with rhIGF-I were those with short stature resulting from defects in growth hormone (GH) action, either due to GH receptor abnormalities or the development of GH-neutralizing antibodies that arose following GH exposure. The administration of rhIGF-I to such children results in improvements in growth rate that are sustained over many years. This appears to improve adult height, but so far the effects are not as robust as when GH-deficient children are treated with human GH. Side effects are common but manageable and seldom necessitate discontinuation of therapy. PMID- 19293581 TI - Phenotypic aspects of growth hormone- and IGF-I-resistant syndromes. AB - Major advances in the diagnosis and characterisation of growth hormone (GH) and IGF-I resistant disorders have occurred during the past 15 years. With these advances has come the realisation that there is broad phenotypic variation within these diagnostic categories. We discuss the current status of endocrine and molecular evaluation, focussing on the phenotypic characteristics of genetic defects in the GH-IGF-I axis. PMID- 19293582 TI - Double diabetes: a mixture of type 1 and type 2 diabetes in youth. AB - The increase in the incidence of type 1 diabetes (T1D), especially in children <5 years of age reported over the past decade can be attributed to changes in environmental factors, either quantitative or qualitative, rather than to an effect of genetic factors operating in such a short period of time. The notable increase in the incidence of type 2 diabetes (T2D) in children and adolescents is very likely the consequence of the increase in obesity and sedentary life style occurring in developed countries. The increase in the number of children and adolescents with a mixture of the two types of diabetes has recently come to light (i.e. subjects who are obese and/or with signs of insulin resistance as well as positive for markers of autoimmunity to beta cells). Under the current classification, it is difficult to define the type of diabetes affecting these young subjects, being classified as T2D because they are obese and insulin resistant, but also as T1D because of the presence of auto-antibodies to beta cells. There is no doubt that these subjects show an overlapping diabetes phenotype typical of both T1D and T2D suggesting that the current classification of diabetes should be revised taking into account this new form of diabetes which has called double diabetes or hybrid diabetes. PMID- 19293583 TI - Cryptorchidism as part of the testicular dysgenesis syndrome: the environmental connection. AB - Cryptorchidism is part of the testicular dysgenesis syndrome (TDS), which includes other male reproductive disorders such as hypospadias, testis cancer and reduced semen quality. These diseases appear to be linked by common pathogenic mechanisms, interfering with normal fetal testis development. Testis development and descent is dependent on androgens and thus on an intact hypothalamus pituitary-gonadal axis. Although cryptorchidism occurs in rare syndromes and genetic disorders, in the majority of children the etiology remains open. Many maternal and fetal risk factors have been previously identified but recently, scientific focus has also been directed to environmental hormone disrupting chemicals and lifestyle, as the prevalence of testis cancer and cryptorchidism has increased and semen quality decreased over few decades in several countries. Some persistent environmental chemicals, e.g. polychlorinated pesticides and polybrominated flame retardants, were associated with testicular maldescent and testis cancer. In addition, prenatal exposure to phthalates was negatively correlated to testosterone levels and anogenital distance as a measure of androgen effect in infant boys. Alcohol consumption and maternal smoking during pregnancy also appeared to be a risk factor for cryptorchidism. Thus, current evidence suggests that the development of the male reproductive tract may be susceptible to adverse effects of environmental hormone disrupters. PMID- 19293584 TI - Disorders of sex development in developmental syndromes. AB - Disorders of sex development comprise an array of congenital conditions with atypical development of chromosomal, gonadal, and anatomical sex affecting the genitourinary tract and in most instances also the endocrine-reproductive system. While the molecular basis of some of these disorders has been well established, it remains elusive in others. This holds true especially for disorders of sex development that are associated with other congenital malformations and abnormalities in a 'syndromic' condition. Syndromic disorders of sex development may be due to monogenic defects, biochemical abnormalities of steroid synthesis, or cytogenetic abnormalities comprising microdeletions or duplications or unbalanced rearrangements. This review will focus on the clinical description of syndromic disorders of sex development guiding towards the genetic classification. The characterization of the underlying diagnosis will improve genetic counseling of the family including prognosis and recurrence risk. PMID- 19293585 TI - In utero treatment of severe fetal anaemia due to parvovirus B19 in one fetus in a twin pregnancy--a case report and literature review. AB - BACKGROUND: Parvovirus B19 (PVB19) is a well-established cause of nonimmune hydrops fetalis and fetal anaemia in pregnancy. However, discordant viral infection of only 1 fetus in a twin pregnancy is a rare occurrence. CASE REPORT: A 40-year-old female with dichorionic, diamniotic twin pregnancy presented at 22 weeks with fetal hydrops and severe anaemia in 1 twin. Maternal PVB19 infection was confirmed, and the affected fetus was treated with a single intrauterine transfusion. The only subsequent complication developed was that the affected fetus was growing on the 5th centile. The affected twin continued to grow, and 2 live twins were delivered by caesarean section at 36 weeks. By 18 months of age, the affected twin had normal development. CONCLUSION: PVB19 may selectively affect 1 fetus in a dichorionic, diamniotic twin pregnancy and may be treated efficiently with intrauterine transfusion. Discordant viral infection in this case suggests that the viral load may be limited to 1 placenta and/or that different individual immunological fetal response starts very early in utero. PMID- 19293586 TI - Prenatal findings in a case of massive fetomaternal hemorrhage associated with intraplacental choriocarcinoma. AB - We describe biochemical assessment of maternal circulation in a case of massive fetomaternal hemorrhage at term associated with intraplacental choriocarcinoma. Markedly elevated maternal serum hCG level at 37 weeks of gestation suggested choriocarcinoma as a cause of fetomaternal hemorrhage in this case. Measurement of maternal hCG may be a useful parameter when intraplacental choriocarcinoma is in the differential diagnosis. In addition, the placenta should be examined in all cases of fetomaternal hemorrhage. PMID- 19293587 TI - Giant pulmonary sequestration: the rare case requiring the EXIT procedure with resection and ECMO. AB - Although most prenatally diagnosed pulmonary sequestrations (PS) are asymptomatic, large lesions are associated with pleural effusions and pulmonary hypoplasia. We present the first reported case of a prenatally diagnosed giant extralobar pulmonary sequestration that required the ex utero intrapartum treatment (EXIT) procedure with resection and extracorporeal membrane oxygenation (ECMO). We discuss the compelling rationale for performing EXIT-resection-ECMO in the setting of a large thoracic mass and anticipated severe respiratory failure at birth. PMID- 19293588 TI - Cerebrovascular blood flow dynamic changes in fetuses with congenital heart disease. AB - OBJECTIVE: The aim of this study was to determine whether the type of congenital heart disease (CHD) or heart function influence fetal cerebrovascular blood flow dynamics. METHODS: Doppler flow velocimetry was performed in the umbilical artery (UA) and middle cerebral artery (MCA), and the ratio of the UA pulsatility index (PI) to the MCA PI (U/C PI) was determined in 45 fetuses with CHD at 20-40 weeks' gestational age. The control group consisted of 275 healthy fetuses matched for gestational age. Individual PI measurements were converted into Z-scores for statistical analysis. RESULTS: Fetuses with CHD (n = 45) had an increased UA PI (p = 0.001) and U/C PI (p < 0.001) when compared to controls. There was no significant difference in the MCA PI between fetuses with CHD (n = 45) and controls (n = 275), while fetuses with CHD complicated by congestive heart failure (CHF) (n = 10) had a decreased MCA PI (p < 0.001) compared to controls. CONCLUSIONS: The lower PIs observed in the MCA of fetuses with CHD complicated by CHF represents a marker of cerebral vasodilation that is due to cerebral hypoxemia and limiting perfusion. The heart function and type of CHD have an impact on fetal cerebrovascular blood flow distribution and this may contribute to the cause of abnormal neurologic development in these fetuses. PMID- 19293589 TI - Comparison of loop electrosurgical conization with one or two passes in high grade cervical intraepithelial neoplasias. AB - The use of loop electrosurgical conization (LEC) for the treatment of large high grade cervical intraepithelial neoplasias (CINs) is often associated with a difficult procedure that results in accidental sample fragmentation, thermal damage and sometimes the presence of positive margins. This study aims to compare LEC that removes the cervical cone in two blocks (anterior and posterior cervical lips - LEC2) with LEC performed with one pass of the loop (LEC1). In a randomized, controlled trial, patients that needed conization due to high-grade CIN were assigned to one of the techniques. There were no differences in terms of age, cone histopathological diagnosis, blood loss, vaginal injuries, stenosis of the cervical os and specimen artifacts. LEC2 required less hemostatic sutures. LEC2 showed no specimen fragmentation, while LEC1 did (0 vs. 5.9%; p = 0.10). As expected, LEC2 samples were heavier (p = 0.01), included a larger ectocervical area (p = 0.001) and, therefore, had a greater volume (p < 0.001) compared to LEC1 samples. The height of the LEC2 specimens was smaller than that of LEC1 specimens (p < 0.001). LEC2 yielded fewer cases of positive margins (12.7%) than LEC1 (33.3%; p = 0.021). We conclude that the LEC2 technique is an effective treatment choice: it is safe for the patient, with better outcomes regarding sample quality than LEC1. Further studies are encouraged regarding this procedure. PMID- 19293590 TI - Guillain-barre syndrome in pregnancy: early diagnosis and treatment is essential for a favorable outcome. AB - Guillain-Barre syndrome (GBS) is an acute or subacute demyelinating polyneuropathy that rarely complicates pregnancy, but increases maternal and perinatal morbidity. A 15-year-old pregnant woman was admitted with ascending paralysis and acellular lumbar puncture suggestive of GBS. She required mechanical ventilation for 26 days, and was treated with intravenous immunoglobulin and corticosteroids. She recovered well and delivered vaginally a healthy newborn at 35 weeks' gestation. The puerperium was complicated by tracheal stenosis which was surgically corrected. PMID- 19293591 TI - Hepatic epithelial hemangioendothelioma in pregnancy. AB - BACKGROUND: Epithelioid hemangioendothelioma (EHE) is a rare vascular neoplasm that affects various tissues including liver, lung, gastrointestinal tract, head and neck, bone, heart and the central nervous system. CASE REPORT: A case of EHE is presented with lung and liver involvement during pregnancy. The patient had an uncomplicated antenatal course and delivery. Because of her asymptomatic presentation and the lack of safe and effective treatment during pregnancy, the decision was made to continue expectant management during her pregnancy. To our knowledge, this is the only published case report on this tumor with lung and liver involvement during pregnancy. CONCLUSION: Expectant management of hepatic EHE is possible during pregnancy and can result in good maternal and fetal outcomes. PMID- 19293592 TI - Different Mechanisms for the Progression of CKD with ACE Gene Polymorphisms. AB - BACKGROUND/AIMS: The blockade of the renin-angiotensin-aldosterone system is the major target of efforts to prevent the progression of chronic kidney disease (CKD). Dual blockade with angiotensin-converting enzyme (ACE) inhibitor and angiotensin II receptor blocker has been reported to show additive renoprotection. However, three types of insertion/deletion (I/D) polymorphism have been reported, and it is unclear whether the dual blockade is effective for all the ACE genotypes. METHODS: We treated 93 CKD patients with or without dual blockade and analyzed the effects on blood pressure (BP), proteinuria, progression of CKD and the relationship to I/D ACE polymorphisms. RESULTS: After long-term medication (average 33 +/- 2 months), BP decreased in all the genotype groups. However, urinary protein excretion decreased only in the II and DI groups (II: -27.1%, DI: -20.5%, DD: +0.8%). In the II and DI groups, amelioration of the progression of renal failure was correlated with reductions in BP and urinary protein excretion. However, the progression rate of renal failure was not correlated with proteinuria in the DD group. CONCLUSION: Proteinuria and BP are key factors for the progression of CKD in II/DI patients, while controlling the BP rather than reducing the proteinuria appears to be crucial in DD patients. PMID- 19293593 TI - Clinical interpretation of reticulocyte hemoglobin content, RET-Y, in chronic hemodialysis patients. AB - BACKGROUND: Iron deficiency is the most common factor associated with erythropoietin (EPO) hyporesponsiveness. Current iron indices are inadequate to demonstrate the status or utility of iron in erythropoiesis. The aims of this study are to investigate the value of the reticulocyte hemoglobin content, RET-Y, in hemodialysis (HD) patients and compare the levels with conventional iron indices. METHODS: HD patients (n = 289) were divided into 4 groups according to serum ferritin (cutoff value 100 ng/ml) and transferrin saturation (TSAT, cutoff value 20%). The RET-Y value, hemogram and biochemical data were determined and compared between groups. Factors associated with RET-Y were examined. RESULTS: The mean RET-Y value was 1,716 +/- 125 AU. Patients with absolute iron deficiency had lower RET-Y levels and mean corpuscular volume (MCV). Patients with functional iron deficiency had a lower reticulocyte production index and serum albumin levels. MCV, mean corpuscular hemoglobin concentration (MCHC) and albumin were independently correlated with the RET-Y level (all p < 0.001). EPO independent patients had low iron indices and low RET-Y levels, but a higher reticulocyte production index and albumin levels were noted. CONCLUSION: RET-Y levels in HD patients were close to that of the normal population. Low RET-Y levels were observed in patients with absolute iron deficiency and also in EPO independent patients with low ferritin and low TSAT. There was a strong association between the serum albumin and RET-Y levels in chronic HD patients. PMID- 19293594 TI - Physical activity, high-density lipoprotein cholesterol subfractions and lecithin:cholesterol acyltransferase in dialysis patients. AB - BACKGROUND/AIMS: Although a low level of high-density lipoprotein cholesterol (HDL-C) is an independent risk factor for atherosclerotic heart disease, the mechanism of HDL-C abnormality in hemodialysis (HD) as well as peritoneal dialysis (PD) patients is not fully understood. The purpose of this study was to investigate the relationship of physical activity with HDL-C subfractions and lecithin:cholesterol acyltransferase activity in HD and PD patients. METHODS: Thirty-five HD and 26 PD patients were studied. Physical activity was estimated as the average number of steps taken per day over 7 days (steps/day). RESULTS: When possible confounding factors were included in the stepwise multiple regression analyses, in HD patients, steps/day was significantly positively related to HDL(2)-C and apolipoprotein (Apo) A-I, while it was significantly positively related to HDL(3)-C in PD patients. When subjects were subdivided into 3 groups according to steps/day, in HD patients, the highest category of steps/day had significantly higher HDL(2)-C and Apo A-I than the lowest category, while such results were not observed in PD patients. CONCLUSION: These results suggest that the associations of physical activity with HDL-C subfractions and Apo A-I that are known in the general population are more pronounced in HD patients than PD patients. PMID- 19293595 TI - Comparison of antibody monitoring system with flow cytometric crossmatch test in renal transplant recipients with high panel-reactive antibody. AB - BACKGROUND/AIMS: The antibody monitoring system (AMS) is a recently developed enzyme-linked immunosorbent assay (ELISA) crossmatch assay to detect donor specific anti-HLA immunoglobulin G antibodies (DS-HLA Abs). This study was conducted to compare the AMS with the flow cytometric crossmatch (FCXM) test in renal transplant recipients with high panel-reactive antibody (PRA). METHODS: Thirty-two sera were obtained from 10 patients with panel reactivity above 50%. When anti-HLA Ab was detected by ELISA PRA and the matched donor had the corresponding HLA antigen, it was considered to indicate DS-HLA Ab. The results of the AMS assay and FCXM were compared with the DS-HLA Abs. RESULTS: Twenty three (71.9%) sera were positive for DS-HLA Abs by ELISA PRA. The AMS assay showed that the number of compatible sera with DS-HLA Abs was 27 (84.4%), and it was significantly concordant (kappa = 0.649, p < 0.0001). For FCXM, the number of compatible sera with DS-HLA Abs was 26 (81.3%), and it was also significantly concordant (kappa = 0.614, p < 0.0001). There was a significant degree of concordance between the AMS assay and FCXM in detection of DS-HLA Abs (kappa = 0.452, p = 0.010). CONCLUSION: The AMS assay is comparable to FCXM in detecting DS-HLA Abs in high PRA recipients. PMID- 19293596 TI - The pathogenic role of Notch activation in podocytes. AB - Podocytes play a key role in the maintenance of the glomerular filtration barrier. Depletion or dysregulative mechanisms of podocytes can lead to the development of glomerulosclerosis. Signaling pathways that control these processes in podocytes are not fully understood. Recent studies from our and other laboratories found that genes that belong to the Notch pathway are regulated in patients and in animal models of renal disease. Genetic studies performed on mice with conditional expression of active Notch1 protein showed massive albuminuria, glomerulosclerosis, and ultimately renal failure and death of the animals. gamma-Secretase inhibitors and genetic deletion of Notch transcriptional binding partner (Rbpj) protected animals from nephrotic syndrome. Further studies are needed to define whether the activation of Notch pathway in podocytes represents a common pathomechanism in glomerular injury, and its potential to be a therapeutic target for the treatment of chronic kidney disease. PMID- 19293597 TI - Metabolic profiles in urine reflect nephrotoxicity of sirolimus and cyclosporine following rat kidney transplantation. AB - BACKGROUND: Cyclosporine and/or sirolimus impair recovery of renal transplants. This study examines the changes in urine metabolite profiles as surrogate markers of renal cell metabolism and function after cyclosporine and/or sirolimus treatment employing a rat kidney transplantation model. METHODS: Using inbred Lewis rats, kidneys were transplanted into bilaterally nephrectomized recipients followed by treatment with either CsA (cyclosporine) 10, Rapa (sirolimus) 1, CsA10/Rapa1 or CsA25/Rapa1 mg/kg/day for 7 days. On day 7, urine was analyzed by (1)H-NMR spectroscopy. Blood and kidney tissue drug concentrations, tissue high energy compounds (including ATP, ADP) and oxidative stress markers (15-F(2t) isoprostanes) in urine were measured by HPLC mass spectrometry. RESULTS: Changes in urine metabolites followed the order Rapa1 < CsA10 < CsA10/Rapa1 < CsA25/Rapa1. Compared with controls, CsA25/Rapa1 showed the greatest changes (creatinine -36%, succinate -57%, citrate -89%, alpha-ketoglutarate -75%, creatine +498%, trimethylamine +210% and taurine +370%). 15-F(2t)-isoprostane concentrations in urine increased in the combined immunosuppressant-treated animals ([CsA25/Rapa1]: 795 +/- 222, [CsA10/Rapa1]: 475 +/- 233 pg/mg/creatinine) as compared with controls (165 +/- 78 pg/mg creatinine). Rapa concentration in blood and tissues increased in the combined treatment (blood: 31 +/- 8 ng/ml, tissue: 1.3 +/- 0.4 ng/mg) as compared with monotherapy (blood: 14 +/- 8 ng/ml, tissue: 0.35 +/- 0.15 ng/mg). Drug blood concentrations correlated with isoprostane urine concentrations, which correlated negatively with citrate, alpha ketoglutarate and creatinine concentrations in urine. Only CsA25/Rapa1 significantly reduced high-energy metabolite concentrations in transplant kidney tissue (ATP -55%, ADP -24%). CONCLUSION: Immunosuppressant drugs induce changes in urine metabolite patterns, suggesting that immunosuppressant-induced oxidative stress is an early event in the development of nephrotoxicity. Urine 15-F(2t) isoprostane concentrations and metabolite profiles may be sensitive markers of immunosuppressant-induced nephrotoxicity. PMID- 19293598 TI - Reduction in VEGF protein and phosphorylated nephrin associated with proteinuria in adriamycin nephropathy rats. AB - BACKGROUND/AIMS: The relationship between vascular endothelial growth factor (VEGF) and the phosphorylated critical podocyte slit diaphragm molecule nephrin is not fully clarified. This study investigated the dynamic changes in VEGF expression and nephrin phosphorylation, and the effects of the antiproteinuric drugs prednisone and lisinopril on them in Adriamycin nephropathy rats. METHODS: Renal tissues from Adriamycin rats were collected at days 3, 7, 14, and 28. Distribution and expression of VEGF was revealed by immunohistochemistry, real time PCR and Western blot. Phosphorylated nephrin was evaluated by immunoprecipitation. RESULTS: A discontinuous redistribution of VEGF was displayed at day 3, followed by significant protein reduction at day 7 with persistent downregulation to day 28. Phosphorylated nephrin decreased evidently at day 14 and persisted to day 28. The reduction in VEGF and phosphorylated nephrin was not a result of podocyte loss. The intervention of prednisone and lisinopril evidently reduced proteinuria, effectively attenuated the severe lesions of podocyte foot processes, and restored the reduction in VEGF and nephrin phosphorylation. At day 28, the reduction in VEGF and phosphorylated nephrin was negatively correlated with proteinuria, whereas the phosphorylated nephrin was positively correlated with VEGF protein from day 7 to day 28. CONCLUSION: The reduction in VEGF protein and nephrin phosphorylation was possibly involved in the proteinuria in Adriamycin rats, and there might be some relationship between VEGF and nephrin phosphorylation. The antiproteinuric effects of lisinopril and prednisone were achieved at least partially by restoring VEGF protein and nephrin phosphorylation. PMID- 19293599 TI - Suppression subtractive hybridization analysis of gene expression during late kidney development identifies the developmentally regulated gene rPEA3. AB - BACKGROUND: While early kidney development has been studied exhaustively, the later stages of nephrogenesis that occur after birth in the rodent are relatively poorly understood. To gain insight into this process, we detected the alterations in gene expression in rat kidney at two postnatal stages, P0 (0 day after birth), the time at which nephrogensis is still active, and P21 (21 days after birth), when nephrogenesis is complete. METHODS: Sprague-Dawley rats were mated, and appearance of a vaginal plug was designated as E0. Kidneys were dissected from embryos at E13, E15, E17 and E19, and from postnatal days P0, P7, P14, P21 and adult rats. Suppression subtractive hybridization (SSH) analysis was performed and highly expressed genes were evaluated as molecular markers by real-time reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, immunofluorescence, and Western blot. RESULTS: Several differentially expressed genes were identified, including rPEA3, a member of the PEA3 subfamily of Ets domain transcription factors. Real time RT-PCR analysis revealed that rPEA3 exhibited dynamic developmental regulation, with high levels of expression from embryonic day E15 until birth, and declining levels thereafter. By in situ hybridization, rPEA3 mRNA was detected in the ureteric bud (UB) and surrounding metanephric mesenchyme of the kidneys from E15 until birth, but was undetectable in mature kidneys. Double-immunofluorescence staining showed that both rPEA3 and WT1 expressed in the condensed mesenchymal cells at E15 and E17; and later from E19 to P7, both expressed in the epithelial cells of ureteric bud and their branches. CONCLUSIONS: These studies provide compelling evidence that SSH is an effective method for identifying genes that are regulated during renal development, and suggest that the newly identified gene rPEA3 may play an important role in kidney development and differentiation. PMID- 19293600 TI - Angiotensin II regulation of renal dopamine uptake and Na(+),K(+)-ATPase activity. AB - BACKGROUND/AIMS: Angiotensin II (ANG II) decreases dopamine (DA) uptake in renal cortex activating AT(1) receptors. We investigated the signaling pathways that mediate this action and the incidence of DA-ANG II interaction on renal Na(+),K(+)-ATPase activity. METHODS: ANG II effects on [(3)H]-DA uptake and Na(+),K(+)-ATPase were measured in samples from the outer renal cortex of Sprague Dawley rats. RESULTS: Inhibition of the phospholipase C (PLC) pathway blunted ANG II inhibitory effects on [(3)H]-DA uptake, since U-73122, 2-APB, TMB-8, chelerythrine and KN-93 (PLC, IP(3)-dependent Ca(2+) release channels, IP(3) receptors, protein kinase C and CaM kinase II inhibitors, respectively) each one blocked ANG II effects. Inhibition of adenylate cyclase pathway did not modify ANG II inhibitory effects on DA uptake. ANG II effects on [(3)H]-DA uptake were able to modify Na(+),K(+)-ATPase activity in carbidopa-treated rats. Exogenous DA decreased while ANG II increased the enzyme activity. Neither the addition of DA together with ANG II, nor the extraneuronal DA uptake blocker hydrocortisone altered ANG II stimulatory effects on Na(+),K(+)-ATPase activity, but hydrocortisone blocked the inhibitory effects of exogenous DA. CONCLUSION: Stimulation of renal AT(1) receptors by ANG II signals through the PLC pathway to inhibit extraneuronal DA uptake. DA and ANG II act through a common pathway involving reversible renal tubular Na(+),K(+)-ATPase deactivation and activation, respectively. In addition, ANG II by itself is able to stimulate renal Na(+),K(+) ATPase activity. PMID- 19293601 TI - Hypopituitarism following Radiotherapy Revisited. AB - Neuroendocrine disturbances in anterior pituitary hormone secretion are common following radiation damage to the hypothalamic-pituitary (H-P) axis, the severity and frequency of which correlate with the total radiation dose delivered to the H P axis and the length of follow-up. The somatotropic axis is the most vulnerable to radiation damage and GH deficiency remains the most frequently seen endocrinopathy. Compensatory hyperstimulation of a partially damaged somatotropic axis may restore normality of spontaneous GH secretion in the context of reduced but normal stimulated responses in adults. At its extreme, endogenous hyperstimulation may limit further stimulation by insulin-induced hypoglycaemia resulting in subnormal GH responses despite the normality of spontaneous GH secretion. In children, failure of the hyper-stimulated partially damaged H-P axis to meet the increased demands for GH during growth and puberty may explain what has previously been described as radiation-induced GH neurosecretory dysfunction and, unlike in adults, the insulin tolerance test remains the gold standard for assessing H-P functional reserve. With low radiation doses (<30 Gy) GH deficiency usually occurs in isolation in about 30% of patients, while with radiation doses of 30-50 Gy, the incidence of GH deficiency can reach 50-100% and long-term gonadotropin, TSH and ACTH deficiencies occur in 20-30, 3-9 and 3-6% of patients, respectively. With higher dose cranial irradiation (>60 Gy) or following conventional irradiation for pituitary tumours (30-50 Gy), multiple hormonal deficiencies occur in 30-60% after 10 years of follow-up. Precocious puberty can occur after radiation doses of <30 Gy in girls only, and in both sexes equally with a radiation dose of 30-50 Gy. Hyperprolactinaemia, due to hypothalamic damage is mostly seen in young women after high dose cranial irradiation and is usually subclinical. H-P dysfunction is progressive and irreversible and can have an adverse impact on growth, body image, sexual function and quality of life. Regular testing is advised to ensure timely diagnosis and early hormone replacement therapy. PMID- 19293602 TI - Alterations in pubertal timing following therapy for childhood malignancies. AB - The onset of puberty marks a time of rapid linear growth, sexual development, and transition from childhood to maturity. The diagnosis and treatment of a childhood malignancy prior to the onset of puberty has the potential to profoundly affect the timing and the tempo of puberty. CNS tumors located in the hypothalamic pituitary (H-P) region, surgical resection in this location, and exposure to CNS radiotherapy are all associated with both precocious and delayed puberty. Also, chemotherapy and radiation can directly damage the gonads, which can result in absent, arrested, or delayed puberty. As a consequence of these alterations of pubertal timing, both male and female survivors of childhood cancer may be at risk of adult short-stature, decreased bone-mineral density, absent or incomplete sexual development, and ultimately, reduced rates of fertility. Appropriate and timely assessment of survivors at high risk of alterations in pubertal development will enable the identification of patients who would benefit from early medical intervention. PMID- 19293603 TI - Obesity during and after Treatment for Childhood Cancer. AB - Obesity is a common complication of treatment for some childhood cancers, particularly acute lymphoblastic leukaemia (ALL) and craniopharyngioma. Evidence based guidance is available for the general paediatric population on the diagnosis, aetiology, consequences, prevention and treatment of obesity, and this should be considered as the starting point for considering such issues in patients with malignancy. In ALL, a high proportion of patients show rapid and excessive weight gain soon after diagnosis which originates partly in lifestyle, in particular via markedly reduced levels of physical activity. Good evidence on risk factors for obesity in ALL is available, and the natural history and aetiology of obesity in ALL are now fairly well understood, while for craniopharyngioma the natural history is reasonably well understood. Understanding the natural history and aetiology of obesity should facilitate preventive interventions in the future. Evidence on preventive interventions is required urgently, and it should focus on promotion of a reduction in sedentary behaviour and increases in physical activity. Such interventions should be helpful in obesity prevention, but could also have a wide range of additional benefits in the prevention or amelioration of other late effects of treatment. PMID- 19293604 TI - Metabolic disorders. AB - Adult survivors of childhood cancer, particularly brain tumours and acute lymphoblastic leukaemia demonstrate evidence of increased rates of metabolic complications and cardiovascular disease in later life. Evidence is accumulating that risk factors for these complications include obesity, physical inactivity, lipid abnormalities, insulin resistance and development of the metabolic syndrome. Cranial radiotherapy-induced growth hormone deficiency, other direct adverse effects of radiotherapy and anthracycline-induced left ventricular dysfunction are clearly identified risk factors for developing these complications. Growth hormone replacement, where appropriate, has been of some benefit in reducing the prevalence of metabolic complications in some long-term survivors. In others, it is clear that multidisciplinary interventions will need to be developed which focus on modifying aspects of lifestyle including increasing levels of habitual physical activity, improving diet and prevention of smoking along with the use of lipid-lowering medication. PMID- 19293605 TI - Bone and bone turnover. AB - Children with cancer are exposed to multiple influences that may adversely affect bone health. Some treatments have direct deleterious effects on bone whilst others may have indirect effects mediated through various endocrine abnormalities. Most clinical outcome studies have concentrated on survivors of acute lymphoblastic leukaemia (ALL). There is now good evidence that earlier treatment protocols that included cranial irradiation with doses of 24 Gy or greater may result in growth hormone deficiency and low bone mineral density (BMD) in the lumbar spine and femoral neck. Under current protocols, BMD decreases during intensive chemotherapy and fracture risk increases. Although total body BMD may eventually return to normal after completion of chemotherapy, lumbar spine trabecular BMD may remain low for many years. The implications for long-term fracture risk are unknown. Risk factors for low BMD include high dose methotrexate, higher cumulative doses of glucocorticoids, male gender and low physical activity. BMD outcome in non-ALL childhood cancers has been less well studied but there is evidence that survivors of childhood brain or bone tumours, and survivors of bone marrow transplants for childhood malignancy, all have a high risk of long-term osteopenia. Long-term follow-up is required, with appropriate treatment of any endocrine abnormalities identified. PMID- 19293606 TI - Male fertility and strategies for fertility preservation following childhood cancer treatment. AB - Infertility in the male is a potential complication of childhood cancer treatment for long-term survivors. The risk is dependent primarily on the treatment used, but also on the underlying disease. Chemotherapy (especially alkylating agents) and radiotherapy, even in low doses, may damage the seminiferous epithelium and impair spermatogenesis in both children and adults. Leydig cell function and testosterone production are generally preserved after chemotherapy and low dose radiotherapy, whilst larger doses of radiotherapy may result in hypogonadism. Patients treated with potentially gonadotoxic treatments require regular multidisciplinary follow-up including assessment of puberty and gonadal function. Currently the only option available for fertility preservation in young males treated for cancer is semen cryopreservation. For pre-pubertal patients, techniques for fertility preservation remain theoretical and as yet unproven. These include hormonal manipulation of the gonadal environment before treatment, germ cell transplantation and testis xenografting, which have all shown promise in a variety of animal studies. Refinement of these techniques requires investigations in relevant animal models. In the present chapter we include data which suggest that the common marmoset (Callithrix jacchus) monkey, a New World primate, exhibits important parallels with human testicular development and may help us to understand why the pre-pubertal testis is vulnerable to effects of cytotoxic therapy on future fertility. PMID- 19293607 TI - Fertility in female childhood cancer survivors. AB - Advances in childhood cancer treatment over the past decades have significantly improved survival, resulting in a rapidly enlarging group of childhood cancer survivors. There is much concern, however, about the effects of treatment on reproductive potential. In women there is evidence that both chemotherapy and radiotherapy may have an adverse effect on ovarian function, ovarian reserve and uterine function, clinically leading to sub-fertility, infertility, premature menopause and/or adverse pregnancy outcomes. Here we will first address normal female fertility and methods to detect decreased fertility. Hence we will focus on direct effects as well as late fertility-related adverse effects caused by chemotherapy and radiotherapy, and we will conclude with a summary of current options for fertility preservation in female childhood cancer survivors. PMID- 19293608 TI - Long-term follow-up of survivors of childhood cancer. AB - Today more than 75% of children treated for cancer will be cured, and attention is focusing on the late effects of treatments for these long-term survivors. Treatment-related morbidity is diverse, with potential effects on the endocrine system (growth, puberty, fertility, pituitary, thyroid and other disorders), cardiovascular, pulmonary and renal complications, second tumours, cognitive, education, neuropsychological and social manifestations. Multi-disciplinary long term follow-up of these patients is essential to monitor, treat, and prevent morbidity. Depending on the nature of the treatment delivered, long-term follow up of the survivor of childhood cancer can be individualised and delivered by a wide range of health professionals either in hospital or in primary care. In this review we describe the chronic health problems encountered by survivors and discuss the development of a long-term follow-up service for childhood cancer survivors. PMID- 19293609 TI - [Correction of complex lower extremity deformities with the use of the Ilizarov Taylor spatial frame]. AB - OBJECTIVES: We evaluated the effectiveness of the Taylor spatial frame (Smith & Nephew, Memphis, TN, USA) in the treatment of complex lower extremity deformities. METHODS: The Taylor spatial frame (TSF) was applied to 29 bone segments of 25 patients (12 females, 13 males; mean age 17 years). Indications for the TSF were congenital disorders (n=12), rickets (n=6), physeal injuries (n=4), stiff nonunions (n=3), malunions (n=3), and sequela from septic arthritis of the knee (n=1). Applications involved the tibia (n=15), femur (n=9), foot (n=4), and knee (n=1) with (n=24) or without (n=5) osteotomies. Following acute correction with the use of the TSF and internal osteosynthesis by plating or nailing, the fixator was removed in six cases. The chronic mode was used in six cases who underwent acute correction. The remaining deformities were gradually corrected using the "total residual mode". The follow-up period ranged from eight months to 42 months (mean 29 months). RESULTS: The mean duration of external fixator was 24.5 weeks (range 18 to 37 weeks) in 13 tibial and five femoral segments. In all cases, correction was applied until the mechanical axis reached normal limits. Complete consolidation was achieved in all osteotomized segments, including three cases of nonunion. A plantigrade foot was obtained in all foot deformities. Recurrence was seen in one case in which knee contracture and subluxation were treated with soft tissue distraction without osteotomy. CONCLUSION: The Taylor spatial frame is a safe and practical method with excellent results in the treatment of nonunions and deformities complicated especially by translation and rotation providing correct clinical data are derived and used. PMID- 19293610 TI - [Treatment of adult tibial diaphysis fractures with reamed and locked intramedullary nailing]. AB - OBJECTIVES: We evaluated the results of reamed and locked intramedullary nailing for tibial diaphysis fractures. METHODS: The study included 73 patients (68 males, 5 females; mean age 31 years; range 17 to 68 years) who were treated with reamed and locked intramedullary nailing for tibial diaphysis fractures. There were 28 AO/ASIF type A, 29 type B, and 16 type C fractures. The fractures involved the proximal 1/3 (n=12), middle 1/3 (n=50), and distal 1/3 (n=11) of the tibial diaphysis. Twenty-eight fractures (38.4%) were closed. According to the Gustilo-Anderson classification, 30 patients (41.1%) had grade I, 10 patients (13.7%) had grade II, and five patients (6.9%) had grade IIIA open fractures. Intramedullary nailing was performed following open reduction in 17 patients (23.3%), and closed reduction in 56 patients (76.7%). The mean time to surgery was 3.4 days (range 2 to 11 days) and the mean follow-up was 48 months (range 24 to 60 months). The patients were evaluated with respect to range of motion, time to union, and complications. Functional results were assessed using the Johner Wrush criteria. RESULTS: Union was achieved in all the patients within a mean of 18.2 weeks (range 8 to 52 weeks). Four patients required dynamization because of delayed union and grafting was performed in one patient. Transient sensorial deficit occurred in one patient after dynamization. One patient underwent revision surgery because of migration of the distal locking screws. The only limitation of range of motion was seen in flexion of two patients (2.7%) who developed anterior knee pain. According to the Johner-Wrush criteria, functional results were very good in 45 patients (61.6%), and good in 28 patients (38.4%). CONCLUSION: Treatment of tibial diaphysis fractures with reamed and locked intramedullary nailing is an appropriate choice with a low complication rate. It can be safely used in moderately contaminated open fractures. PMID- 19293611 TI - [Comparison between locked intramedullary nailing and plate osteosynthesis in the management of adult forearm fractures]. AB - OBJECTIVES: We evaluated the results of two different surgical methods for the treatment of adult diaphyseal fractures of both forearm bones. METHODS: Forty-two adult patients with forearm fractures were retrospectively evaluated. Of these, 22 patients (7 women, 15 men; mean age 32 years; range 18 to 69 years) underwent open reduction and plate-screw fixation, and 20 patients (6 women, 14 men; mean age 33 years; range 18 to 70 years) underwent closed reduction and locked intramedullary nail fixation. The fractures were classified according to the AO/OTA system. The patients were assessed using the Grace-Eversmann criteria and the DASH (Disability of the Arm, Shoulder and Hand) questionnaire. The mean follow-up was 30 months (range 12 to 45) with plate-screw fixation, and 23 months (range 12 to 34) with intramedullary nailing. RESULTS: The mean operation time was 65 minutes (range 40 to 97 min) with plate-screw fixation, and 61 minutes (range 35 to 90 min) with intramedullary nailing (p>0.05). The mean time to union was significantly shorter with intramedullary nailing (10 weeks vs. 14 weeks; p<0.05). According to the Grace-Eversmann criteria, the results were excellent or good in 18 patients (81.8%) and acceptable in four patients (18.2%) treated with plate-screw fixation, compared to 18 patients (90%) and two patients (10%), respectively, treated with intramedullary nailing. The mean DASH scores were 15 (range 4 to 30) and 13 (range 3 to 25), respectively. The two groups did not differ significantly with respect to functional results and DASH scores (p>0.05). Postoperative complications were seen in three patients (13.6%) and two patients (10%) with plate-screw fixation and intramedullary nailing, respectively. CONCLUSION: The two fixation methods yield similar results in terms of functional healing and patient satisfaction in the management of adult forearm fractures. PMID- 19293612 TI - [Functional results of the parallel-plate technique for complex distal humerus fractures]. AB - OBJECTIVES: We evaluated functional results of patients treated with open reduction and internal fixation with the parallel-plate technique for complex distal humerus fractures. METHODS: Twenty-one patients (14 males, 7 females; mean age 47 years; range 16 to 85) underwent open reduction with olecranon osteotomy and internal fixation with the parallel-plate technique for distal humerus fractures accompanied by highly intra-articular or metaphyseal comminution (n=10), intra-articular comminution and osteoporosis (n=7), and intra-articular and metaphyseal comminution with bone loss (n=4). According to the AO classification, there were 12 C3, six C2, and three C1 type fractures. Eight patients had open fractures. The mean time to surgery was six days (range 1 to 17 days). Functional results were evaluated using the Mayo elbow performance score, Jupiter elbow score, and DASH (Disabilities of the Arm, Shoulder and Hand) score. The mean follow-up was 28 months (range 12 to 48 months). RESULTS: The mean total range of motion was 90.2+/-31.1 degrees, flexion was 118.1+/-17.4 degrees, and extension was 27.8+/-17.4 degrees. The mean Mayo elbow performance score and DASH score were 86.1+/-12.6 and 7.6+/-9.5, respectively. According to the Jupiter elbow scores, the results were excellent in seven patients, good in 11 patients, moderate in two patients, and poor in one patient. Radiographically, solid union was achieved in all the patients. Heterotopic ossification of varying degrees was seen in seven patients, two of whom underwent resection of heterotopic ossification due to severe limitation of movement. Debridement was performed in one patient due to the development of deep infection. Chondrolysis of the elbow occurred in one patient. Patients with open fractures had significantly lower range of motion than those with closed fractures (p<0.05), but the Mayo elbow performance score and DASH score did not differ significantly in this respect (p>0.05). CONCLUSION: Functional results are satisfactory in distal humerus fractures treated with stable osteosynthesis and parallel-plate technique that allow early active motion. PMID- 19293613 TI - [The effect of resistance-related proteins on the prognosis and survival of patients with osteosarcoma: an immunohistochemical analysis]. AB - OBJECTIVES: Despite the developments in chemotherapy protocols, improvement in the survival rates of osteosarcoma has been limited. We evaluated the effect of certain prognosis-related proteins on survival of patients with osteosarcoma. METHODS: Data from 45 patients (24 males, 21 females) who were treated and followed-up for osteosarcoma were reviewed. Following neoadjuvant chemotherapy, 41 patients underwent extremity saving surgery, and four patients underwent amputation. The most frequent localization was the lower end of the femur (n=23, 51.1%), followed by the upper end of the tibia (n=10, 22.2%). Three patients had metastasis on admission. Surgical resection samples were retrieved from the pathology archive and analyzed immunohistochemically for the expression of p glycoprotein p170, p53, heat-shock protein 27 (HSP27), HSP90, and nm23. The effect of these proteins on prognosis and survival was assessed with survival analysis using the Kaplan-Meier method. The mean follow-up was 49.7 months (range 6 to 185 months). RESULTS: Three patients with metastasis on admission died within five years due to pulmonary metastasis. New metastases developed in 29 patients. Total 5-year and 10-year survival rates were 60% and 43%, respectively. The corresponding disease-free survival rates were 41% and 24%. Five-year survival was 29% in patients who developed metastasis. Among clinical factors, survival was influenced only by the presence of metastasis on admission (p=0.044). Five-year and 10-year survival rates were significantly different between patients with and without p53 positivity (p=0.04), while the other proteins were not significantly associated with survival. CONCLUSION: Our data suggest that p53 may be used as a prognostic marker in osteosarcoma due to its significant association with survival. PMID- 19293614 TI - [The importance of patient selection for the treatment of proximal humerus fractures with percutaneous technique]. AB - OBJECTIVES: We evaluated the indications and disadvantages of percutaneous technique for proximal humerus fractures in relation to complications encountered in osteoporotic elderly patients and the importance of patient selection. METHODS: The study included 18 patients (10 men, 8 women; mean age 48 years; range 14 to 89 years) who underwent percutaneous fixation (closed reduction and pin fixation with K-wires or Schanz screws) for proximal humerus fractures. Eight patients were beyond 60 years of age. According to the Neer classification, five patients had two-part, 13 patients had three-part neck fractures. The patients were evaluated with range of motion of the shoulder, radiographs, and the Disability of Arm Shoulder and Hand questionnaire (DASH). The mean follow-up was 23 months (range 8 to 60 months). RESULTS: The mean shoulder abduction was 134 degrees (range 30 degrees to 160 degrees) and the mean elevation was 118 degrees (range 30 degrees to 140 degrees). Full range of motion of the shoulder was achieved in 11 patients (61.1%), including all with two-part fractures. Abduction losses of 60 degrees to 130 degrees and less than 30 degrees were seen in four patients and three patients, respectively, all of whom were older than 60 years. The mean DASH score was 18 (range 0 to 77). Fourteen patients having a score of less than 10 had no pain or functional complaints. Four patients with a score of more than 10 were older than 70 years. Pin migration was observed in seven patients (38.9%), all of whom were over 60 years of age. One patient required revision with partial prosthesis. Nonunion was seen in one patient (5.6%) and malunion with a varus/valgus deformity occurred in four patients (22.2%). None of the patients developed avascular necrosis. CONCLUSION: Percutaneous fixation may be preferred in the treatment of two-part and carefully-selected three-part proximal humerus fractures. Due to high complication rate, patient selection is of primary importance among elderly patients. PMID- 19293615 TI - [The effect of ankle position on intracompartmental pressures of the leg]. AB - OBJECTIVES: We investigated differences in the intracompartmental pressures (ICP) of the leg in relation to various positions of the ankle joint in patients with chronic exertional compartment syndrome (CECS). METHODS: The study included 16 patients (10 males, 6 females; mean age 30+/-9 years, range 16 to 48 years) actively involved in various sports. Intracompartmental pressures were monitored with the use of slit catheters connected to a pressure transducer in 28 anterior and 14 deep posterior compartments before and after exercise during the following positions of the ankle joint: relaxed-resting, passive plantar flexion, neutral, and passive dorsiflexion. Alterations in ICP were assessed with reference to that measured in the relaxed-resting position of the ankle. RESULTS: Significant increases in ICP were observed in both anterior and deep posterior compartments during dorsiflexion of the ankle, being 9.1+/-10.6 mmHg (p=0.0001) and 8+/-10.3 mmHg (p=0.001) in the anterior compartment, and 6.4+/-4.4 mmHg (p=0.0001) and 7.2+/-4.3 mmHg (p=0.001) in the deep posterior compartment before and after exercise, respectively. No significant increases were found in other positions of the ankle (p>0.05). While the lowest values of ICP were noted in the relaxed resting position, plantar flexion of the ankle was associated with decreased ICP pressures. CONCLUSION: Dorsiflexion of the ankle increases ICP significantly in both anterior and deep posterior compartments. The results of this study may have clinical implications for the conservative management of both CECS and tibial fractures. PMID- 19293616 TI - [Correlation of basic motor skills with arthroscopic experience]. AB - OBJECTIVES: One of the methods in motor skill teaching is to furnish the students with the individual skills drawn from the target procedure. This method requires identification and defining of all components of the target procedure. This study aimed to define basic motor skills composing arthroscopic skillfulness. METHODS: A total of 42 orthopedists (mean age 38+/-8 years) were enrolled. The study group was comprised of 17 experienced orthopedists working at least for 10 years as a specialist and performing more than 50 arthroscopic procedures per year. The control group included 25 young orthopedists or residents having an arthroscopic experience of less than three years. All the participants were assessed simultaneously and in the same experimental setting. Each participant was tested after having been shown to use in vitro skill development instruments simulating arthroscopic basic motor skills. RESULTS: Compared to the control group, the experienced group had significantly higher mean age (42 vs. 34.4 years), longer duration of arthroscopic experience (12.4 vs. 1.6 years), and greater number of the arthroscopies performed per year (93.9 vs. 26.9) (p=0.000). The mean anticipation time (p=0.028) and two-arm coordination time (p=0.043) were significantly shorter in the experienced group. In correlation analysis, duration of arthroscopic experience was correlated with the mean anticipation time (r= 0.41, p=0.008) and two-arm coordination time (r=-0.33, p=0.033). In addition, the mean anticipation time decreased significantly as the number of arthroscopies increased (r=-0.446, p=0.003). CONCLUSION: Some basic motor skills correlate with arthroscopic competence. The use of these motor skill instruments in arthroscopy training may aid to improve arthroscopic skills. PMID- 19293617 TI - [The effect of low-molecular-weight heparin on rat tendon healing]. AB - OBJECTIVES: We investigated the effect of low-molecular-weight heparin (LMWH) on the healing of tendons. METHODS: Forty-five adult Wistar rats weighing 300 g were randomized into three groups equal in number. All the rats underwent full thickness surgical incision of the Achilles tendon followed by primary repair. After the operation, two groups received daily subcutaneous LMWH injections (nadroparin calcium) for four weeks at high or low doses (group I, 6 mg/kg, 170 IU AXa; group II, 3 mg/kg, 85 IU AXa). Group III remained untreated as the control group. Histologically, the specimens were examined under light and electron microscopy with regard to the amount of fibrillar collagen synthesis, mitochondrial degeneration, and the composition of the extracellular matrix collagen. Biomechanically, maximum load to failure and correspondent elongation of the tendons were measured. RESULTS: Compared to the control group, histologically, both LMWH-treated groups exhibited increased number of fibroblasts, increased fibrillar collagen formation in the extracellular matrix, and higher counts of granular endoplasmic reticula in cytoplasmic contents of fibroblasts as well as decreased mitochondrial vacuolization and degeneration. Biomechanical assessments showed that tendons in group I had significantly higher maximum load to failure and elongation values than group II and III (31 N vs. 24.6 N and 23.1 N; 25 mm vs. 19.6 mm and 17.3 mm, respectively; p<0.05). Groups II and III did not differ significantly in this respect (p>0.05). CONCLUSION: Daily administration of single dose LMWH improves tendon healing through increasing the number of fibroblasts and fibrillar collagen synthesis and decreasing mitochondrial degeneration. PMID- 19293618 TI - [Anconeus arthroplasty: a salvage procedure in recurrent heterotopic ossification]. AB - Anconeus interpositional arthroplasty has been used in the treatment of radiocapitellar and radioulnar joint problems occurring after trauma. We performed anconeus interposition arthroplasty in a 31-year-old male patient who developed heterotopic ossification (HTO) in the elbow following surgical treatment of an isolated radial neck fracture. Treatment with implant removal and excision of the radial head resulted in recurrent HTO and a stiff elbow. The patient underwent excision of the heterotrophic new bone and mobilization of the proximal radius. The anconeus muscle was mobilized and interposed between the proximal radius and ulna. Sixteen months after the operation, forearm rotation, elbow flexion and extension increased by 80 degrees , 45 degrees , and 60 degrees , respectively. Control X-rays showed maintenance of the radiohumeral gap without any signs of HTO. PMID- 19293619 TI - [Knee ochronotic arthropathy and arthroscopic findings]. AB - Ochronotic arthropathy is a rare condition found in patients with alkaptonuria which is a hereditary metabolic disease associated with deposition of homogentisic acid derivatives in the articular cartilage, menisci, ligaments, and connective tissues due to homogentisic acid oxidase deficiency. These pigmentary changes are termed ochronosis. We presented a 50-year-old woman in whom arthroscopic examination of the right knee revealed brown-black discoloration of the articular cartilage and menisci leading to the diagnosis of alkaptonuria by further laboratory examinations. PMID- 19293620 TI - [Osteochondritis dissecans occurring in the patella and medial femoral condyle in the same knee]. AB - Osteochondritis dissecans (OCD) is a condition in which a portion of articular cartilage separates from the bone together with the underlying subchondral bone. Its classical localization is the medial femoral condyle. We presented a 14-year old boy who had OCD lesions in both the medial femoral condyle and patella in the same knee joint. He presented with complaints of pain, swelling, and locking following sports activities. Magnetic resonance imaging showed completely detached loose bodies causing instability. Arthroscopic treatment was performed including in situ fixation of the condylar lesion and debridement of patellar lesions. At six-month follow-up, he had full range of motion of the knee joint, with some pain in the patellar grind test. PMID- 19293621 TI - [Treatment algorithm for patients with puberphonia]. AB - OBJECTIVES: We evaluated the results of treatment for puberphonia and aimed to develop a treatment algorithm for patients with puberphonia. PATIENTS AND METHODS: Sixteen male patients (mean age 21.5 years; range 16 to 34 years) with puberphonia underwent voice therapy (3-10 sessions). Perceptual and acoustic analyses of vocal quality were performed in 12 patients. Perceptual analysis included the Voice Handicap Index (VHI) and videolaryngostroboscopy (VLS) and acoustic evaluations included F0 (fundamental frequency), jitter, shimmer, and NNE (normalized noise energy). RESULTS: Following voice therapy, all scores of the VHI showed significant improvements (p=0.001). There was a significant improvement in vibratory pattern and mucosal wave of vocal cords in VLS evaluation (p=0.004 and p=0.002, respectively). Among acoustic parameters, only the mean F0 showed a significant change from 246 Hz to 134 Hz after treatment (p=0.001). Stabilization of F0 could not be achieved in two patients, one of whom underwent type III thyroplasty. CONCLUSION: The main difficulties encountered in the treatment of puberphonia include stabilization of the attained F0 and widening the frequency range. Implementation of the treatment algorithm through a step-by-step approach provides an objective way of assessing the disease and its management. PMID- 19293622 TI - [Head and neck paragangliomas]. AB - OBJECTIVES: We evaluated patients who were treated for head and neck paragangliomas. PATIENTS AND METHODS: The study included 40 patients (25 females, 15 males; mean age 48 years; range 26 to 74 years) who were operated on for paragangliomas of the head and neck region between 1993 and 2007. Clinical findings, treatment modalities, and the results of treatment were evaluated. RESULTS: The most common complaint was neck swelling (n=30), followed by tinnitus (n=7), hearing loss (n=6), imbalance (n=3), pain (n=2), hoarseness (n=2), and nasal obstruction (n=1). The mean duration of symptoms was 22 months. The most common paraganglioma was glomus caroticum (n=28) with a mean tumor diameter of 4.5 cm (range 2 to 12 cm). Urinary vanilmandelic acid concentration was measured in 24 patients and found above normal range in two patients. Octreotide scintigraphy was performed in 14 patients and femoral angiography was performed in 27 patients. Multicentric disease was present in one patient and one patient had bilateral involvement. Transcervical excision was the most common approach. Complications were as follows: transient facial nerve paresis (n=3), vagal nerve palsy (n=2), hypoglossal nerve palsy (n=2), permanent facial paralysis (n=1), bleeding (n=1), and total hearing loss (n=1). No recurrences were encountered during a mean follow-up of 71 months. CONCLUSION: Preoperative evaluation of all patients with respect to catecholamine secretion and multicentric disease is important for choosing the proper treatment and preventing possible complications. PMID- 19293623 TI - [Pharyngocutaneous fistula after total laryngectomy: incidence, predisposing factors, and treatment]. AB - OBJECTIVES: We investigated the incidence and etiologic factors of pharyngocutaneous fistulas occurring after total laryngectomy. PATIENTS AND METHODS: The study included 53 patients (46 males, 7 females; mean age 59 years; range 34 to 84 years) who underwent total laryngectomy for epidermoid carcinoma. Risk factors for pharyngocutaneous fistula, its incidence and time of occurrence, treatment approaches, and length of hospital stay were evaluated. RESULTS: Pharyngocutaneous fistula developed in 19 patients (35.9%) within a mean of 7.4 days postoperatively. Age, smoking, the presence of chronic systemic disease; T1 T2 stage, localization, and differentiation of tumor, concomitant bilateral neck dissection, T/Y shaped pharyngeal closure, and high fever in the first postoperative 48 hours did not show any significant relationship with the formation of fistula. Factors significantly associated with fistula formation were T4 stage, alcohol consumption, postoperative anemia, and hypoproteinemia. The mean hospital stay was 40.5 days in patients with pharyngocutaneous fistula compared with 15.3 days in patients without fistula. CONCLUSION: Postoperative anemia, hypoproteinemia, alcohol consumption, and T4 stage are significant risk factors for pharyngocutaneous fistula formation. PMID- 19293624 TI - [Internal jugular vein thrombosis following functional and selective neck dissections]. AB - OBJECTIVES: Internal jugular vein (IJV) thrombosis is a rare complication of functional and selective neck dissections. It increases morbidity and may seldom be fatal. We investigated the frequency of IJV thrombosis and its relationship with the dissection technique. PATIENTS AND METHODS: We evaluated 52 functional and selective neck dissections performed in 34 male patients (mean age 57 years; range 34 to 76 years) with head and neck cancer. Dissections were mainly performed by sharp dissection (n=27) or cautery (n=25). The patients were examined by Doppler ultrasonography with respect to IJV flow and thrombosis preoperatively, and at two weeks and at 3 to 6 months postoperatively. RESULTS: In the early postoperative period, thrombosis was observed in 7.4% (n=2) of the necks treated with sharp dissection and in 4% (n=1) of the necks treated with cautery. There was no statistically significant difference between the two groups with respect to IJV thrombosis. Late Doppler examinations showed complete recanalization of all thrombosed IJVs. CONCLUSION: Our data suggest that sharp dissection or cautery techniques performed in functional neck dissections do not differ with respect to the frequency of postoperative IJV thrombosis. PMID- 19293625 TI - Long-term quality of life evaluation after laser microsurgery with or without adjuvant radiotherapy for laryngeal carcinoma. AB - OBJECTIVES: We evaluated long-term quality of life of patients successfully treated with either laser-assisted microsurgery alone or combined with adjuvant radiotherapy for laryngeal carcinoma. PATIENTS AND METHODS: The study included 53 patients who were treated by laser surgery and 16 patients who were treated by laser surgery combined with adjuvant radiotherapy. Quality of life was evaluated with two validated questionnaires: the global EORTC QLQ-C30 and the head and neck specific EORTC QLQ-H&N35. The mean follow-up was 59 months (range 10 to 111 months). RESULTS: Patients receiving adjuvant radiotherapy showed a significantly lower level of global health status. Surprisingly, the two treatment groups showed close similarities with respect to symptoms specific to the head and neck region. The most frequent complaints were coughing and speech problems in both groups. On the other hand, patients with a follow-up duration of more than or less than five years did not differ significantly with regard to the global health status. CONCLUSION: Adjuvant radiotherapy must be selectively utilized in patients treated with laser surgery for laryngeal carcinoma. PMID- 19293626 TI - [Donor site sequela of class III B vascular anomaly following reconstruction of the mandible with free osteoseptocutaneous fibula flap: a case report]. AB - Free transfer of the fibula for mandibular reconstruction provides desirable functional and aesthetic results. However, unexpected donor- and recipient-site complications may be encountered with most free flap transfer procedures. A 62 year-old woman underwent wide resection and segmental mandibulectomy for low grade squamous cell carcinoma of intraoral location. Reconstruction of the defect was performed at the same session with an osteoseptocutaneous free flap from the right fibula. A class III B vascular malformation was detected intraoperatively at the donor site, which resulted in restriction of foot inversion and abduction, and of toe extension during the follow-up. PMID- 19293627 TI - A case of naso-ophthalmic myiasis. AB - Myiasis is defined as an occupational disease and it is mainly seen in people who are in contact with animals, in particular sheep. We detected nasal and external ophthalmomyiasis caused by Oestrus ovis in a 33-year-old male who worked as a research assistant in the faculty of agriculture. The presenting complaints were severe foreign body sensation and discharge in the nose, cough, and foreign body sensation in the right eye. Ophthalmologic examination revealed many moving larvae in the right eye of the patient. Larvae were removed and nasolacrimal duct irrigation was performed through the inferior canaliculus. Endoscopic nasal examination also showed larvae colonies. The larvae were removed under local anesthesia and were taken to the parasitology laboratory, wherein Oestrus ovis was identified. Minimal endoscopic sinus surgery showed nonspecific chronic inflammation. PMID- 19293628 TI - [A giant retropharyngeal lipoma showing no change in clinical presentation and size within a two-year follow-up: a case report]. AB - Retropharyngeal tumors are rare. They typically originate from the neighboring anatomical structures or from the retropharyngeal space itself. Retropharyngeal lipomas are even rarer. A 75-year-old male patient presented with complaints of dyspnea, dysphagia, snoring, and obstructive sleep apnea. Computed tomography showed a giant mass measuring 125 x 81 x 27 mm in the retropharyngeal space, compressing the larynx, trachea, and esophagus, and extending to the thoracic inlet. The mass had regular margins, exhibited low attenuation, and was homogeneously hypodense. After intravenous contrast administration, there were thin septa formations within the lesion that did not show contrast enhancement. There was no invasion to the neighboring muscular structures. The lesion was considered a lipoma. Surgery was not performed due to the cardiovascular problems of the patient. During a two-year follow-up, there were no changes in the size and tissue characteristics of the mass, nor any mass-related clinical problems. PMID- 19293629 TI - [Castleman's disease in a patient with recurrent reactive lymphadenopathy]. AB - Castleman's disease is an uncommon benign disorder associated with progressive lymph node enlargement. It most commonly involves the mediastinum and involvement of the neck accounts for 6%. A 28-year-old male presented with a progressively enlarging left-sided neck mass. He had a history of excisional lymph node biopsy that yielded a histopathologic diagnosis of reactive lymphadenopathy. In our clinic, a repeat excisional biopsy was performed and the diagnosis was made as Castleman's disease. Although it is an uncommon cause of lymphadenopathy in the neck, Castleman's disease should be considered in the differential diagnosis of recurring lymphadenopathies. Repeated excisional biopsies may be necessary (from the largest lymph node when possible) to uncover the disease. PMID- 19293630 TI - Cavernous hemangioma of the nasal bone: reconstruction with cartilage graft. AB - A 32-year-old woman presented with nasal obstruction on the right side and a cosmetic defect one year after surgery for nasal hemangioma. On inspection, the right nasal cavity was partly obstructed by a proliferative mass. Computed tomography showed a 2 x 3-cm soft tissue mass that destroyed the nasal bone. The mass was excised completely with the surrounding tissue and the defect was reconstructed with cartilage taken from the cavum conchae. Cartilage graft is appropriate for reconstruction of nasal bone defects because of its easy availability, elasticity, vitality, and good functional and aesthetic results. PMID- 19293631 TI - [Surgical approach to encapsulated follicular variant of papillary thyroid carcinoma: a report of three cases]. AB - The diagnosis of follicular variant of thyroid papillary carcinoma is often difficult with fine-needle aspiration biopsy since this variant contains both benign and malignant follicular structures. The encapsulated follicular variant poses even more difficulty for diagnosis. It is usually agreed that this variant has similar clinical behavior to that of classic thyroid papillary carcinoma. However, some recent cases have been shown to have an aggressive course, leading to controversy as to the type of surgery. We presented three female patients whose results of fine-needle aspiration biopsy were not malignant, but postoperative histopathologic diagnosis was made as encapsulated follicular variant. One patient underwent thyroid lobectomy for a single thyroid nodule, and two patients underwent subtotal thyroidectomy. Upon histologic diagnosis of encapsulated follicular variant, total thyroidectomy was performed in two patients. Encapsulated follicular variant should be borne in mind in patients with a benign fine-needle aspiration biopsy result. PMID- 19293632 TI - Tie2 is tied at the cell-cell contacts and to extracellular matrix by angiopoietin-1. AB - Angiopoietin-1 (Ang1) binds to and activates Tie2 receptor tyrosine kinase. Ang1 Tie2 signal has been proposed to exhibit two opposite roles in the controlling blood vessels. One is vascular stabilization and the other is vascular angiogenesis. There has been no answer to the question as to how Tie2 induces two opposite responses to the same ligand. Our group and Dr. Alitalos group have demonstrated that trans-associated Tie2 at cell-cell contacts and extracellular matrix (ECM)-anchored Tie2 play distinct roles in the endothelial cells. The complex formation depends on the presence or absence of cell-cell adhesion. Here, we review how Ang1-Tie2 signal regulates vascular maintenance and angiogenesis. We further point to the unanswered questions that must be clarified to extend our knowledge of vascular biology and to progress basic knowledge to the treatment of the diseases in which Ang1-Tie2-mediated signal is central. PMID- 19293633 TI - Sensory involvement in the SOD1-G93A mouse model of amyotrophic lateral sclerosis. AB - A subset of patients of amyotrophic lateral sclerosis (ALS) present with mutation of Cu/Zn superoxide dismutase 1 (SOD1), and such mutants caused an ALS- like disorder when expressed in rodents. These findings implicated SOD1 in ALS pathogenesis and made the transgenic animals a widely used ALS model. However, previous studies of these animals have focused largely on motor neuron damage. We report herein that the spinal cords of mice expressing a human SOD1 mutant (hSOD1 G93A), besides showing typical destruction of motor neurons and axons, exhibit significant damage in the sensory system, including Wallerian-like degeneration in axons of dorsal root and dorsal funiculus, and mitochondrial damage in dorsal root ganglia neurons. Thus, hSOD1-G93A mutation causes both motor and sensory neuropathies, and as such the disease developed in the transgenic mice very closely resembles human ALS. PMID- 19293634 TI - Inhibition of cell proliferation by a resveratrol analog in human pancreatic and breast cancer cells. AB - Resveratrol has been reported to possess cancer preventive properties. In this study, we analyzed anti-tumor activity of a newly synthesized resveratrol analog, cis-3,4',5-trimethoxy-3'-hydroxystilbene (hereafter called 11b) towards breast and pancreatic cancer cell lines. 11b treatments reduced the proliferation of human pancreatic and breast cancer cells, arrested cells in the G2/M phase, and increased the percentage of cells in the subG1/G0 fraction. The 11b treatments also increased the total levels of mitotic checkpoint proteins such as BubR1, Aurora B, Cyclin B, and phosphorylated histone H3. Mechanistically, 11b blocks microtubule polymerization in vitro and it disturbed microtubule networks in both pancreatic and breast cancer cell lines. Computational modeling of the 11b tubulin interaction indicates that the dimethoxyphenyl group of 11b can bind to the colchicine binding site of tubulin. Our studies show that the 11b treatment effects occur at lower concentrations than similar effects associated with resveratrol treatments and that microtubules may be the primary target for the observed effects of 11b. These studies suggest that 11b should be further examined as a potentially potent clinical chemotherapeutic agent for treating pancreatic and breast cancer patients. PMID- 19293635 TI - Selective addition of CXCR3(+) CCR4(-) CD4(+) Th1 cells enhances generation of cytotoxic T cells by dendritic cells in vitro. AB - Increasing importance is being given to the stimulation of Th1 response in cancer immunotherapy because its presence can shift the direction of adaptive immune responses toward protective immunity. Based on chemokine receptor expression, CXCR3(+) CCR4(-) CD4(+) T cells as Th1-type cells were investigated its capacity in monocyte-derived dendritic cell (DC) maturation and polarization, and induction of antigen specific cytotoxic T lymphocytes (CTL) in vitro. The levels of IL-4, IL-5 and IL-10 were decreased to the basal level compared with high production of IFN-gamma, TNF-alpha, and IL-2 in CXCR3+CCR4-CD4+ T cells stimulated with anti-CD3 and anti-CD28 antibodies. Co-incubation of activated CD4(+) or CXCR3(+) CCR4-CD4(+) T cells with DC (CD4(+/) DC or CXCR3(+) CD4(+/) DC, respectively) particularly up-regulated IL-12 and CD80 expression compared with DC matured with TNF-a and LPS (mDC). Although there was no significant difference between the effects of the CXCR3(+) CCR4(-) CD4(+) and CD4(+) T cells on DC phenotype expression, CXCR3(+) CD4(+/) DC in CTL culture were able to expand number of CD8(+) T cells and increased frequencies of IFN-gamma secreting cells and overall cytolytic activity against tumor antigen WT-1. These results demonstrated that the selective addition of CXCR3(+) CCR4(-) CD4(+) T cells to CTL cultures could enhance the induction of CTLs by DC in vitro, and implicated on a novel strategy for adoptive T cell therapy. PMID- 19293636 TI - Resveratrol inhibits foam cell formation via NADPH oxidase 1- mediated reactive oxygen species and monocyte chemotactic protein-1. AB - Resveratrol is a polyphenolic compound in red wine that has anti-oxidant and cardioprotective effects in animal models. Reactive oxygen species (ROS) and monocyte chemotactic protein-1 (MCP-1) play key roles in foam cell formation and atherosclerosis. We studied LPS-mediated foam cell formation and the effect of resveratrol. Resveratrol pretreatment strongly suppressed LPS-induced foam cell formation. To determine if resveratrol affected the expression of genes that control ROS generation in macrophages, NADPH oxidase 1 (Nox1) was measured. Resveratrol treatment of macrophages inhibited LPS-induced Nox1 expression as well as ROS generation, and also suppressed LPS-induced MCP-1 mRNA and protein expression. We investigated the upstream targets of Nox1 and MCP-1 expression and found that Akt-forkhead transcription factors of the O class (FoxO3a) is an important signaling pathway that regulates both genes. These inhibitory effects of resveratrol on Nox1 expression and MCP-1 production may target to the Akt and FoxO3a signaling pathways. PMID- 19293637 TI - Reactive oxygen species regulate the generation of urokinase plasminogen activator in human hepatoma cells via MAPK pathways after treatment with hepatocyte growth factor. AB - Tumor cells are known to produce larger amounts of reactive oxygen species (ROS) than normal cells. Although numerous reports have indicated the importance of ROS in urokinase plasminogen activator (uPA) production, the precise mechanisms remain controversial. In our study, we investigated the effect of ROS on uPA generation in human hepatoma cells, HepG2 and Hep 3B. We determined the effects of hepatocyte growth factor (HGF) on the regulation of ROS, which resulted in suppression of ROS production, as measured with the fluorescent probe, 2'-7' dichlorofluorescein diacetate. The role of HGF in modulating ROS production, particularly that regulated by Rac-1, was determined. HGF suppressed the increment in Rac-1-regulated ROS in both cell lines. Treatment with 200 micrometer of H(2)O(2) showed a 1.6-2.1 fold increment in HGF, but a little increment occurred at 500 micrometer of H(2)O(2). It looks no dose dependent manner. Combined treatment with H(2)O(2) and HGF, resulted in a slightly increased production of HGF compared to no treatment (control). Also, H(2)O(2) upregulated uPA expression in both hepatoma cell lines. To identify the downstream pathways regulated by ROS, we treated cells with PD 98059, an MEK inhibitor, and SB 203580, a p38 inhibitor, after treatment with H(2)O(2), and showed negative control between ERK and p38 kinase activities for uPA regulation. We found that HGF modulate Rac-1-regulated ROS production through activation of Akt and ROS regulates uPA production via MAP kinase, which provides a novel clue to clarify the mechanism underlying hepatoma progression. PMID- 19293638 TI - Langerhans cell protein 1 (LCP1) binds to PNUTS in the nucleus: implications for this complex in transcriptional regulation. AB - Protein phosphatase-1 (PP1) nuclear targeting subunit (PNUTS), also called PP1R10, p99, or CAT 53 was originally isolated as a mammalian nuclear PP1-binding protein. In this study, we performed yeast two-hybrid screens to identify PNUTS interacting proteins. Here, we report that LCP1 (epidermal Langerhans cell protein 1), a novel member of the HMG-box protein family, binds tightly to PNUTS. Co-immunoprecipitation of deletion constructs revealed that the C-terminus of LCP1 is sufficient for the interaction with an N-terminal region of PNUTS that is distinct from its PP1-binding domain. Furthermore, immunofluorescence studies showed that a subpopulation of LCP1 co-localizes with PNUTS in nuclear speckles. Importantly, we found that the N-terminus of LCP1 has a strong trans-activation activity in a GAL4-based heterologous transcription assay. The transcriptional activity of LCP1 is markedly suppressed by its interaction with PNUTS, in a PP1 independent manner. These findings suggest that the coordinated spatial and temporal regulation of LCP1 and PNUTS may be a novel mechanism to control the expression of genes that are critical for certain physiological and pathological processes. PMID- 19293639 TI - AMP kinase/cyclooxygenase-2 pathway regulates proliferation and apoptosis of cancer cells treated with quercetin. AB - AMPK (AMP-activated protein kinase) is highly conserved in eukaryotes, where it functions primarily as a sensor of cellular energy status. Recent studies indicate that AMPK activation strongly suppresses cell proliferation in non malignant cells as well as in tumor cells. In this study, quercetin activated AMPK in MCF breast cancer cell lines and HT-29 colon cancer cells, and this activation of AMPK seemed to be closely related to a decrease in COX-2 expression. The application of a COX-2 inhibitor or cox-2-/- cells supported the idea that AMPK is an upstream signal of COX-2, and is required for the anti proliferatory and pro-apoptotic effects of quercetin. The suppressive or growth inhibitory effects of quercetin on COX-2 were abolished by treating cancer cells with an AMPK inhibitor Compound C. These results suggest that AMPK is crucial to the anti-cancer effect of quercetin and that the AMPK-COX-2 signaling pathway is important in quercetin-mediated cancer control. PMID- 19293640 TI - B cell activation factor (BAFF) is a novel adipokine that links obesity and inflammation. AB - B cell activation factor (BAFF) is a novel member of the TNF ligand superfamily, mainly produced by myeloid cells. BAFF has been shown to participate in B-cell survival and B- and T-cell maturation. BAFF expression in adipocytes has been recently demonstrated. In the current study, we verified that BAFF expression is increased during adipocyte differentiation. BAFF expression was augmented by TNF alpha treatment and was decreased by rosiglitazone treatment. BAFF secretion in lean and in ob/ob mice sera were compared and smaller amount of BAFF was secreted in ob/ob mice. mRNA and protein expression were different between epididymal and visceral adipose tissue. BAFF expression was also increased in ob/ob mouse adipose tissue. We sought to identify known BAFF receptors (BAFF-R, BCMA, and TACI) in adipocytes, and determined that all three were present and upregulated during adipocyte differentiation. However, the expression of TACI was distinct from that of BAFF-R and BCMA under TNF-alpha and BAFF ligand treatment. BAFF-R and BCMA expression levels were upregulated under pro-inflammatory conditions, but TACI was reduced. Conversely, BAFF-R and BCMA expression levels were downregulated by rosiglitazone treatment, but TACI was increased. Taken together, our results suggest that BAFF may be a new adipokine, representing a link between obesity and inflammation. PMID- 19293666 TI - Body mass index, metabolic syndrome and carotid atherosclerosis. AB - OBJECTIVE: Body fatness and fat distribution are widely accepted as coronary heart disease risk factors. In this study, we have evaluated the contribution of generalized adiposity, assessed by body mass index (BMI), to carotid atherosclerosis, in participants with or without metabolic syndrome (MetS). METHODS: We have analysed 840 female and 1002 male participants in a regional Cardiovascular Disease Prevention Campaign. Blood glucose and lipids were analysed by standard methods. According to BMI, calculated as weight (in kilograms)/height (in square metres), participants were divided into normal weight (BMI <25 kg/m2), overweight (BMI between 25 and 29.9 kg/m2) and obese (BMI>29.9 kg/m2). Carotid atherosclerosis was evaluated by echo Doppler. RESULTS: Blood pressure, waist circumference, triglycerides and glucose were significantly higher, and high-density lipoprotein was lower, in overweight and obese participants, compared with normal weight. MetS was more frequent among obese and overweight than normal-weight participants (51.7 vs. 21.5 vs. 9.8%, respectively). The prevalence of carotid atherosclerosis was 45.29% in participants with MetS, significantly higher than in participants without MetS (33.04%, P<0.0001), but it was similar across the three weight categories. Furthermore, in multiple regression analyses BMI was not significantly associated with carotid atherosclerosis. CONCLUSION: The present findings suggest that increasing body weight favours the clustering of coronary heart disease risk factors. Overweight and obesity, however, do not independently associate with carotid atherosclerosis. PMID- 19293667 TI - Hemostatic changes and clinical sequelae after on-pump compared with off-pump coronary artery bypass surgery: a prospective randomized study. AB - OBJECTIVE: To delineate the effects of extracorporeal bypass on biomarkers of hemostasis, fibrinolysis, and inflammation and clinical sequelae. METHODS: Patients were assigned prospectively and randomly to either on-pump (n=41) or off pump (n=51) coronary bypass surgery. The concentrations of C-reactive protein, fibrinogen, D-dimer, and plasminogen activator inhibitor type-1 in blood were quantified before and after (1 and 24 h) surgery. Similar surgical and anesthetic procedures were used for both groups. Clinical events were assessed during initial hospitalization and at the end of 1 year. RESULTS: The concentrations of plasminogen activator inhibitor type-1 and D-dimer were greater compared with preoperative values 1 and 24 h after surgery in both groups, but their concentrations increased to a greater extent 24 h after surgery in the on-pump group (P<0.01). The concentration of C-reactive protein did not change appreciably immediately after surgery in either group but increased in a parallel manner 24 h after either on-pump or off-pump surgery (P<0.01). Bypass surgery in the on-pump group was associated with greater blood loss during surgery and more bleeding after surgery (P< or =0.01). The incidence of all other complications was similar in the two groups. CONCLUSION: On-pump surgery was associated with biochemical evidence of a prothrombotic state early after surgery but no greater incidence of thrombotic events was observed. The prothrombotic state might be a consequence of extracorporeal bypass, compensation in response to more bleeding, or both in patients undergoing on-pump surgery. PMID- 19293668 TI - Increased circulating placental growth factor during percutaneous coronary intervention is associated with applied radiocontrast agent. AB - BACKGROUND AND AIM OF THE STUDY: Recent studies have suggested placental growth factor (PlGF) and vascular endothelial growth factor (VEGF) as promising new biomarkers for risk stratification in acute coronary syndromes (ACS). However, little is known about the influence of percutaneous coronary intervention (PCI) on circulating PlGF and VEGF levels. METHODS: Thirty-five patients with ACS, 27 patients with stable coronary artery disease (sCAD), and nine healthy controls were enrolled in the study. Although all patients with ACS and 14 patients with stable angina pectoris underwent PCI, 13 patients with coronary artery disease required no revascularization (sCAD). PlGF and VEGF plasma concentrations were measured by immunoassay during and at the end of PCI and coronary angiography. RESULTS: Plasma PlGF levels were comparable in patients with ACS and sCAD on admission. Although coronary angiography or heparin alone did not alter PlGF and VEGF levels, immediately after PCI a dramatic increase was seen in circulating PlGF and a decrease in VEGF, which was independent of the clinical presentation of the patients, heparin administration, or the angiographic procedure itself, but was associated with the extent of coronary artery disease and the amount of the injected contrast media. In-vitro experiments revealed that radiocontrast agents induced the release of PlGF from endothelial cells without altering PlGF mRNA expression. CONCLUSION: Patients undergoing PCI exhibit an increase in circulating PlGF, probably caused by posttranslational modifications of radiocontrast agents in endothelial cells. Therefore, analysis of plasma PlGF and VEGF levels may consider the timing of blood sampling with respect to PCI and contrast media exposure. PMID- 19293669 TI - Safety and efficacy of 2.5-mm sirolimus-eluting stent implantation at lower deployment pressures in very small vessels (<2.5 mm). AB - OBJECTIVES: Percutaneous coronary interventions for very small vessels are common in clinical practice despite an unavailability of the 2.25-mm sirolimus-eluting stent (SES) in some countries. We sought to evaluate the clinical and angiographic outcomes of 2.5-mm SES implantation at lower deployment pressures in very small coronary arteries. METHODS: Between June 2004 and March 2007, a total of 244 patients underwent percutaneous coronary interventions in vessels with reference diameters less than 2.5 mm at our centers: outcomes in 126 consecutive patients undergoing 2.5-mm SES implantation at lower deployment pressures (< or =10 atmospheres) with predilatation and postdilatation were compared with those in 118 patients who received bare-metal stents (BMS). RESULTS: In the SES group, rates of predilatation and postdilatation were 73.8 and 81% respectively, and mean deployment pressure was 8.3+/-1.2 atmospheres. At follow-up, in-segment late loss was markedly lower in SES versus BMS (0.21+/-0.41 vs. 0.48+/-0.63 mm, P=0.001), resulting in significantly lower rates of restenosis (14.7 vs. 37.5%, P<0.001). At 1 year, SES versus BMS use was associated with similar rates of stent thrombosis (0.8 vs. 0.8%, P>0.999), but significantly lower rates of major adverse cardiac events (MACE) (11.9 vs. 27.1%, P=0.003), mainly driven by a significantly lower need for target-lesion revascularization (9.5 vs. 26.3%, P=0.001). Multivariable analysis identified the SES use as independently associated with a reduced 1-year MACE risk (hazard ratio: 0.32; 95% confidence interval: 0.15-0.66; P=0.002). CONCLUSION: Implantation of 2.5-mm SES in vessels with reference diameters less than 2.5 mm using lower deployment pressures and predilatation and postdilatation may lead to reduced risks of restenosis and MACE without an increased risk of stent thrombosis up to 1 year. PMID- 19293670 TI - Bibliography--editors' selection of current world literature. PMID- 19293671 TI - Clefts and craniofacial surgery: innovations and contributions in motion. PMID- 19293673 TI - Festschrift to honor John B. Mulliken, MD. Guest editors' introduction. PMID- 19293676 TI - Judah Folkman, MD, 1933-2008: father of angiogenesis. PMID- 19293677 TI - Remembering Dr Judah Folkman. PMID- 19293678 TI - Pharmacological treatment of a diffuse arteriovenous malformation of the upper extremity in a child. AB - A young girl with an arteriovenous malformation involving the right upper extremity developed rapidly progressive bony destruction that did not respond to embolization. Treatment with marimastat, starting at 3 years of age, resulted in rapid resolution of pain and gradual healing of bony destruction, associated with regression of the intraosseous arteriovenous shunts. New arteriovenous shunts with bony destruction developed over the years and responded to an increase in the dose of marimastat. Interruption of therapy resulted in recurrence of pain and formation of new lesions. The patient has been treated in this way for 12 years with no adverse effects from the drug. PMID- 19293679 TI - Sense of wonder. PMID- 19293680 TI - Perspectives on craniosynostosis: sutural biology, some well-known syndromes, and some unusual syndromes. AB - Perspectives on craniosynostosis are discussed under the following headings: sutural biology (anatomic and genetic categories of synostosis; sutures, suture systems, and types of craniosynostosis; well-known syndromes (Muenke syndrome and Pfeiffer syndrome); and unusual syndromes (thanatophoric dysplasia, Beare Stevenson cutis gyrata syndrome, Crouzonodermoskeletal syndrome, Carpenter syndrome, Elejalde syndrome, hypomandibular faciocranial syndrome, and craniorhiny). Five of these syndromes are caused by fibroblast growth factor receptor (FGFR) mutations; one is caused by ras-like in rat brain 23 (RAB23) mutations; and three have Mendelian patterns of inheritance, but the molecular basis remains unknown to date. PMID- 19293681 TI - Cloverleaf skulls: etiologic heterogeneity and pathogenetic variability. AB - Cloverleaf skulls (Kleeblattschadel-Syndromen, trilobular skulls) have different modes of development (pathogenetic variability). Synostosis may involve various sutures in different types of cloverleaf skulls. Their facial features, radiographic findings, and CNS anomalies are also reviewed. Different causes of cloverleaf skulls have been identified (etiologic heterogeneity). One table lists 26 different conditions with trilobular skulls identified under various etiologic headings: monogenic, chromosomal, disruptive, iatrogenic, and distinctive conditions of unknown cause. Discussed in detail are type 2 thanatophoric dysplasia, type 2 Pfeiffer syndrome, Apert syndrome, amniotic bands, Say Poznanski syndrome, and COH syndrome. Another table summarizes the relative frequencies of the cloverleaf skull syndromes. PMID- 19293683 TI - Nonunion after fracture of the anterior tibial spine: case report and review of the literature. AB - Nonunion of anterior tibial spine of tibia in children is quite rare, but it could be associated with significant instability of the knee as it involves the fixation of anterior cruciate ligament. We report one case in which open reduction and internal fixation was carried out with good functional results. A literature review was performed to identify the cases reported of tibial eminence nonunion in children. PMID- 19293685 TI - Updated outcomes of partner notification for human immunodeficiency virus, San Francisco, 2004-2008. AB - An evaluation of HIV-partner notification demonstrated its effectiveness in identifying new cases of HIV infection in San Francisco. Findings suggested that health departments should consider focusing efforts on reducing the time between diagnosis and interview. Such data are critical for the prioritization of HIV case-finding activities and enable local health departments to measure the costs of partner services, as well as identify areas for quality improvement. PMID- 19293686 TI - Identification of human immunodeficiency virus-1 specific CD8+ and CD4+ T cell responses in perinatally-infected infants and their mothers. AB - BACKGROUND: There are few data describing the specificity, breadth and magnitude of T cell responses to HIV-1 in infancy. METHODS: HIV-specific CD8+ and CD4+ T cell responses to peptide pools representing Gag, Env, Pol, Nef and the regulatory regions (Reg) were simultaneously measured in 18 perinatally-infected infants and 14 of their chronically-infected mothers, using a whole blood interleukin-2 and interferon-gamma flow cytometric intracellular cytokine staining assay. RESULTS: HIV-specific CD8+ T cell responses were detected in all the infants aged 6 weeks and older (range 0.1-6.62%) and their mothers (range 0.1 4.89%). HIV-specific CD4+ T cell responses were detected in 33% of the infants (range 0.11-0.54%) and 73% of the mothers (range 0.16-0.84). CD8+ T cell responses in the mothers were almost equally spread between the variable (Nef, Reg and Env) and conserved proteins (Gag and Pol). Conversely, CD8+ T cell responses to the more variable proteins dominated in the perinatally-infected infants comprising 74% of the total response. Interestingly, mothers and infants shared responses to at least one peptide pool, whereas only one mother-infant pair shared a peptide pool targeted by CD4+ T cells. Two in-utero-infected infants tested at birth had CD8+ T cell responses, and one of them had an Env specific CD4 T cell response. CONCLUSION: Our observations that HIV-specific CD8+ and CD4+ T cell responses can be detected in perinatally-infected infants from 6 weeks of age and that CD8+ T cell responses predominantly target the variable proteins have important implications for HIV vaccine design. PMID- 19293687 TI - Severe retropharyngeal abscess after the use of a reinforced laryngeal mask with a Bosworth introducer. PMID- 19293688 TI - The LMA CTrach in patients with difficult airways. PMID- 19293689 TI - Obstructive sleep apnea of obese adults: pathophysiology and perioperative airway management. AB - Collapsible pharyngeal airway size is determined by interaction between structural properties of the pharyngeal airway and neural regulation of the pharyngeal dilating muscles. Obesity seems to have two distinct mechanical influences on the pharyngeal airway collapsibility. First, obesity increases soft tissue surrounding the pharyngeal airway within limited maxillomandible enclosure occupying and narrowing its space (pharyngeal anatomical imbalance). Second, obesity, particularly central obesity, increases visceral fat volume decreasing lung volume. Pharyngeal wall collapsibility is increased by the lung volume reduction, possibly through decreased longitudinal tracheal traction (lung volume hypothesis). Neural compensation for functioning structural abnormalities operating during wakefulness is lost during sleep, leading to pharyngeal obstruction. Instability of the negative feedback of the respiratory system may accelerate cycling of pharyngeal closure and opening. Improvement of the pharyngeal anatomical imbalance and maintenance of lung volume are the keys for safe perioperative airway managements of obese patients with obstructive sleep apnea. PMID- 19293690 TI - Comparison of two insertion techniques of ProSeal laryngeal mask airway: standard versus 90-degree rotation. AB - BACKGROUND: This study compared two insertion techniques of ProSeal laryngeal mask airway. METHODS: A total of 160 female patients (American Society of Anesthesiologists physical status I or II; age 18-80 yrs) undergoing gynecologic surgery were randomly allocated to the standard or rotational technique groups. In the standard technique group (n = 80), ProSeal laryngeal mask airway insertion was performed by a single experienced user using digital manipulation. In the rotational technique group (n = 80), the ProSeal laryngeal mask airway was rotated counter clockwise through 90 degrees in the mouth and advanced until the resistance of the hypopharynx was felt, and then straightened out in the hypopharynx (n = 80). The ease of insertion was assessed by the success rate at the first attempt. Heart rate and mean blood pressure were recorded 1 min before and 1 min after insertion. Postoperative complications were noted. RESULTS: The success rate of insertion at the first attempt was higher for the rotational technique (100% vs. 85%, P < 0.001). The overall success rate, i.e., successful insertion within three attempts, was 94% for the standard technique versus 100% for the rotational technique. There was no significant change in heart rate, but mean blood pressure increased significantly with the standard technique (P = 0.001). The incidence of blood staining (9% vs. 36%, P < 0.001) and sore throat (8% vs. 25%, P = 0.005) was lower with the rotational technique. CONCLUSION: The rotational technique is more successful than the standard technique and is associated with less pharyngeal mucosal trauma, as evidenced by a lower incidence of sore throat and mucosal bleeding. PMID- 19293691 TI - Prediction and outcomes of impossible mask ventilation: a review of 50,000 anesthetics. AB - BACKGROUND: There are no existing data regarding risk factors for impossible mask ventilation and limited data regarding its incidence. The authors sought to determine the incidence, predictors, and outcomes associated with impossible mask ventilation. METHODS: The authors performed an observational study over a 4-yr period. For each adult patient undergoing a general anesthetic, preoperative patient characteristics, detailed airway physical exam, and airway outcome data were collected. The primary outcome was impossible mask ventilation defined as the inability to exchange air during bag-mask ventilation attempts, despite multiple providers, airway adjuvants, or neuromuscular blockade. Secondary outcomes included the final, definitive airway management technique and direct laryngoscopy view. The incidence of impossible mask ventilation was calculated. Independent (P < 0.05) predictors of impossible mask ventilation were identified by performing a logistic regression full model fit. RESULTS: Over a 4-yr period from 2004 to 2008, 53,041 attempts at mask ventilation were recorded. A total of 77 cases of impossible mask ventilation (0.15%) were observed. Neck radiation changes, male sex, sleep apnea, Mallampati III or IV, and presence of beard were identified as independent predictors. The receiver-operating-characteristic area under the curve for this model was 0.80 +/- 0.03. Nineteen impossible mask ventilation patients (25%) also demonstrated difficult intubation, with 15 being intubated successfully. Twelve patients required an alternative intubation technique, including two surgical airways and two patients who were awakened and underwent successful fiberoptic intubation. CONCLUSIONS: Impossible mask ventilation is an infrequent airway event that is associated with difficult intubation. Neck radiation changes represent the most significant clinical predictor of impossible mask ventilation in the patient dataset. PMID- 19293692 TI - Three-dimensional morphological analyses of positional dependence in patients with obstructive sleep apnea syndrome. AB - BACKGROUND: It is generally considered that patients with obstructive sleep apnea syndrome (OSAS) at increased perioperative risk should be placed in nonsupine positions throughout the recovery process; however, not all patients with OSAS show positional dependence. The authors hypothesized that morphological differences exist in three-dimensional (3D) soft tissue and craniofacial structures of the pharyngeal airway between positional and nonpositional OSAS. METHODS: The subjects of the study were body mass index-matched, age-matched, and apnea hypopnea index-matched positional (n = 10) and nonpositional (n = 10) Japanese OSAS patients and body mass index-matched Japanese control subjects (n = 10). Pharyngeal magnetic resonance imaging and cephalometric radiography were performed during wakefulness. RESULTS: The patients with positional OSAS had a smaller volume of the pharyngeal lateral wall soft tissues, larger maxilla-nasion mandible angle, and smaller lower facial height than the nonpositional OSAS and the control subjects. The patients with positional OSAS showed a significantly steeper sella-nasion-mandible angle and smaller craniofacial volume than the control subjects. There were no significant differences in tongue volume and 3D pharyngeal anatomical balance between positional and nonpositional OSAS. Multivariate stepwise regression for positional dependence showed that the dominant determinant was the volume of the lateral pharyngeal wall, followed by lower facial height and maxilla-nasion-mandible angle. CONCLUSIONS: Patients with positional OSAS have wider airways in the lateral parts, lower facial height, and more backward position of the lower jaw, which may explain differences in the maintenance of pharyngeal airway patency in the lateral sleep position. PMID- 19293693 TI - Continuous positive airway pressure via the Boussignac system immediately after extubation improves lung function in morbidly obese patients with obstructive sleep apnea undergoing laparoscopic bariatric surgery. AB - BACKGROUND: Morbidly obese patients are at elevated risk of perioperative pulmonary complications, including airway obstruction and atelectasis. Continuous positive airway pressure may improve postoperative lung mechanics and reduce postoperative complications in patients undergoing abdominal surgery. METHODS: Forty morbidly obese patients with known obstructive sleep apnea undergoing laproscopic bariatric surgery with standardized anesthesia care were randomly assigned to receive continuous positive airway pressure via the Boussignac system immediately after extubation (Boussignac group) or supplemental oxygen (standard care group). All subjects had continuous positive airway pressure initiated 30 min after extubation in the postanesthesia care unit via identical noninvasive ventilators. The primary outcome was the relative reduction in forced vital capacity from baseline to 24 h after extubation. RESULTS: Forty patients were enrolled into the study, 20 into each group. There were no significant differences in baseline characteristics between the groups. The intervention predicted less reduction in all measured lung functions: forced expiratory volume in 1 s (coefficient 0.37, SE 0.13, P = 0.003, CI 0.13-0.62), forced vital capacity (coefficient 0.39, SE 0.14, P = 0.006, CI 0.11-0.66), and peak expiratory flow rate (coefficient 0.82, SE 0.31, P = 0.008, CI 0.21-0.1.4). CONCLUSIONS: Administration of continuous positive airway pressure immediately after extubation maintains spirometric lung function at 24 h after laparoscopic bariatric surgery better than continuous positive airway pressure started in the postanesthesia care unit. PMID- 19293694 TI - Identification of patients at risk for postoperative respiratory complications using a preoperative obstructive sleep apnea screening tool and postanesthesia care assessment. AB - BACKGROUND: Patients with obstructive sleep apnea are at risk for perioperative morbidity. The authors used a screening prediction model for obstructive sleep apnea to generate a sleep apnea clinical score (SACS) that identified patients at high or low risk for obstructive sleep apnea. This was combined with postanesthesia care unit (PACU) monitoring with the aim of identifying patients at high risk of postoperative oxygen desaturation and respiratory complications. METHODS: In this prospective cohort study, surgical patients with a hospital stay longer than 48 h who consented were enrolled. The SACS (high or low risk) was calculated; all patients were monitored in the PACU for recurrent episodes of bradypnea, apnea, desaturations, and pain-sedation mismatch. All patients underwent pulse oximetry postoperatively; complications were documented. Chi square, two-sample t test, and logistic regression were used for analysis. The oxygen desaturation index (number of desaturations per hour) was calculated. Oxygen desaturation index and incidence of postoperative cardiorespiratory complications were primary endpoints. RESULTS: Six hundred ninety-three patients were enrolled. From multivariable logistic regression analysis, the likelihood of a postoperative oxygen desaturation index greater than 10 was increased with a high SACS (odds ratio = 1.9, P < 0.001) and recurrent PACU events (odds ratio = 1.5, P = 0.036). Postoperative respiratory events were also associated with a high SACS (odds ratio = 3.5, P < 0.001) and recurrent PACU events (odds ratio = 21.0, P < 0.001). CONCLUSIONS: Combination of an obstructive sleep apnea screening tool preoperatively (SACS) and recurrent PACU respiratory events was associated with a higher oxygen desaturation index and postoperative respiratory complications. A two-phase process to identify patients at higher risk for perioperative respiratory desaturations and complications may be useful to stratify and manage surgical patients postoperatively. PMID- 19293695 TI - Lithium protects against anesthesia-induced developmental neuroapoptosis. AB - BACKGROUND: Ethanol and anesthetic drugs trigger neuroapoptosis in the developing mouse brain. Recently, it was found that ethanol-induced neuroapoptosis is preceded by suppressed phosphorylation of extracellular signal-regulated protein kinase (ERK), and lithium counteracts both the phosphorylated ERK suppressant action and ethanol-induced neuroapoptosis. The current study was undertaken to address the following questions. (1) Do ketamine and propofol mimic ethanol in suppressing ERK phosphorylation? (2) If they do, does lithium prevent this suppressant action and also prevent these anesthetic drugs from triggering neuroapoptosis? METHOD: Postnatal day 5 mice were treated with propofol, ketamine, lithium, a combination of propofol or ketamine with lithium or saline, and their brains were prepared for Western blot analysis or histology. For Western blot, cytosolic lysates of caudate putamen were analyzed for expression of phosphorylated ERK and phosphorylated serine/threonine-specific protein kinase. For histology, brains were stained immunohistochemically with antibodies to activated caspase-3, and the density of activated caspase-3 positive cells was determined. RESULTS: Ketamine and propofol suppressed phosphorylated ERK, and lithium counteracted both the phosphorylated ERK suppressant action and neuroapoptotic action of these anesthetic drugs. CONCLUSION: If further testing finds lithium to be safe for use in pediatric/obstetric medicine, administration of a single dose of lithium before anesthesia induction may be a suitable means of mitigating the risk of anesthesia-induced developmental neuroapoptosis. PMID- 19293696 TI - Effect of hypercarbia and isoflurane on brain cell death and neurocognitive dysfunction in 7-day-old rats. AB - BACKGROUND: Millions of neonates undergo anesthesia each year. Certain anesthetic agents cause brain cell death and long-term neurocognitive dysfunction in postnatal day (P)7 rats. Despite its intuitive appeal, a causal link between cell death and neurocognitive decline after anesthesia has not been established. If one existed, the degree of cell death would be expected to correlate with the degree of neurocognitive dysfunction caused by anesthesia. The authors therefore tested if cell death caused by various durations of isoflurane at 1 minimum alveolar concentration causes duration-dependent long-term neurocognitive dysfunction. METHODS: Isoflurane was administered to P7 rats at 1 minimum alveolar concentration for 0, 1, 2, or 4 h. To control for the respiratory depressant effects of anesthesia, a group of rats was treated with 4 h of carbon dioxide. Cell death was assessed by FluoroJade staining 12 h after the end of each intervention, and neurocognitive outcome was assessed 8 weeks later by using fear conditioning, spatial reference memory, and spatial working memory tasks. RESULTS: Widespread brain cell death was caused by 2 h and 4 h of isoflurane and by 4 h of carbon dioxide. The degree and distribution of thalamic cell death was similar in 4 h isoflurane-treated and 4-h carbon dioxide-treated rats. Only 4 h of isoflurane caused a long-term neurocognitive deficit affecting both spatial reference memory and spatial working memory. Working memory was improved in carbon dioxide-treated rats. CONCLUSION: Isoflurane-induced brain cell death may be partly caused by hypercarbia. The inconsistencies between cell death and neurocognitive outcome suggest that additional or alternative mechanisms may mediate anesthesia-induced long-term neurocognitive dysfunction. PMID- 19293697 TI - Isoflurane inhibits growth but does not cause cell death in hippocampal neural precursor cells grown in culture. AB - BACKGROUND: Isoflurane causes long-term hippocampal-dependent learning deficits in rats despite limited isoflurane-induced hippocampal cell death, raising questions about the causality between isoflurane-induced cell death and isoflurane-induced cognitive function. Neurogenesis in the dentate gyrus is required for hippocampal-dependent learning and thus constitutes a potential alternative mechanism by which cognition can be altered after neonatal anesthesia. The authors tested the hypothesis that isoflurane alters proliferation and differentiation of hippocampal neural progenitor cells. METHODS: Multipotent neural progenitor cells were isolated from pooled rat hippocampi (postnatal day 2) and grown in culture. These cells were exposed to isoflurane and evaluated for cell death using lactate dehydrogenase release, caspase activity, and immunocytochemistry for nuclear localization of cleaved caspase 3. Growth was assessed by cell counting and BrdU incorporation. Expression of markers of stemness (Sox2) and cell division (Ki67) were determined by quantitative polymerase chain reaction. Cell fate selection was assessed using immunocytochemistry to stain for neuronal and glial markers. RESULTS: Isoflurane did not change lactate dehydrogenase release, activity of caspase 3/7, or the amount of nuclear cleaved caspase 3. Isoflurane decreased caspase 9 activity, inhibited proliferation, and decreased the proportion of cells in s-phase. messenger ribonucleic acid expression of Sox2 (stem cells) and Ki67 (proliferation) were decreased. Differentiating neural progenitor cells more often select a neuronal fate after isoflurane exposure. CONCLUSIONS: The authors conclude that isoflurane does not cause cell death, but it does act directly on neural progenitor cells independently of effects on the surrounding brain to decrease proliferation and increase neuronal fate selection. These changes could adversely affect cognition after isoflurane anesthesia. PMID- 19293698 TI - Inhibition of p75 neurotrophin receptor attenuates isoflurane-mediated neuronal apoptosis in the neonatal central nervous system. AB - BACKGROUND: Exposure to anesthetics during synaptogenesis results in apoptosis and subsequent cognitive dysfunction in adulthood. Probrain-derived neurotrophic factor (proBDNF) is involved in synaptogenesis and can induce neuronal apoptosis via p75 neurotrophic receptors (p75). proBDNF is cleaved into mature BDNF (mBDNF) by plasmin, a protease converted from plasminogen by tissue plasminogen activator (tPA) that is released with neuronal activity; mBDNF supports survival and stabilizes synapses through tropomyosin receptor kinase B. The authors hypothesized that anesthetics suppress tPA release from neurons, enhance p75 signaling, and reduce synapses, resulting in apoptosis. METHODS: Primary neurons (DIV5) and postnatal day 5-7 (PND5-7) mice were exposed to isoflurane (1.4%, 4 h) in 5% CO2, 95% air. Apoptosis was assessed by cleaved caspase-3 (Cl-Csp3) immunoblot and immunofluorescence microscopy. Dendritic spine changes were evaluated with the neuronal spine marker, drebrin. Changes in synapses in PND5-7 mouse hippocampi were assessed by electron microscopy. Primary neurons were exposed to tPA, plasmin, or pharmacologic inhibitors of p75 (Fc-p75 or TAT-Pep5) 15 min before isoflurane. TAT-Pep5 was administered by intraperitoneal injection to PND5-7 mice 15 min before isoflurane. RESULTS: Exposure of neurons in vitro (DIV5) to isoflurane decreased tPA in the culture medium, reduced drebrin expression (marker of dendritic filopodial spines), and enhanced Cl-Csp3. tPA, plasmin, or TAT-Pep5 stabilized dendritic filopodial spines and decreased Cl-Csp3 in neurons. TAT-Pep5 blocked isoflurane-mediated increase in Cl-Csp3 and reduced synapses in PND5-7 mouse hippocampi. CONCLUSION: tPA, plasmin, or p75 inhibition blocked isoflurane-mediated reduction in dendritic filopodial spines and neuronal apoptosis in vitro. Isoflurane reduced synapses and enhanced Cl-Csp3 in the hippocampus of PND5-7 mice, the latter effect being mitigated by p75 inhibition in vivo. These data support the hypothesis that isoflurane neurotoxicity in the developing rodent brain is mediated by reduced synaptic tPA release and enhanced proBDNF/p75-mediated apoptosis. PMID- 19293699 TI - Behavior and development in children and age at the time of first anesthetic exposure. AB - BACKGROUND: Several experimental studies have suggested that early exposure to anesthetic agents, i.e., before completion of synaptogenesis, can result in widespread apoptotic neuronal degeneration and late cognitive impairment, but human data are lacking. The authors performed a retrospective pilot study to test the feasibility and calculate sample sizes for a larger epidemiologic study of disturbed neurobehavioral development as a function of age at the time of first anesthetic exposure. Pediatric urological procedures were selected because the timing of surgery depends mainly on the age at which a diagnosis is made. METHODS: Neurobehavioral development was assessed using the validated 120-item parental Child Behavior CheckList/4-18 in 314 children who were operated for pediatric urological procedures between the ages of 0 and 6 yr. RESULTS: Of 243 questionnaires returned, the total problem score was clinically deviant in 41 (23%) of children aged less than 24 months at the time of first surgery and 13 (20%) aged greater than 24 months. Crude and adjusted odds ratios for a clinically deviant Child Behavior CheckList/4-18 score increased with younger age at the time of surgery, but the confidence intervals were very wide. Adjusted odds ratio was 1.38 (0.59-3.22) when operated at age less than 6 months, 1.19 (0.45-3.18) when operated between 6 and 12 months of age, and 1.20 (0.45-3.20) when operated between 12 and 24 months (using operated at greater than 24 months of age as reference category). A properly powered cohort study would require at least 2,268 children. CONCLUSIONS: Children undergoing urologic surgery at age less than 24 months showed more behavioral disturbances than children in whom surgery was performed after age 2 yr, although the results were not statistically significant. To confirm or refute an effect of anesthesia on cognitive development, at least 2,268 children need to be studied. With retrospective study designs, residual confounding remains an issue that can only be solved in prospective randomized studies. PMID- 19293700 TI - Early exposure to anesthesia and learning disabilities in a population-based birth cohort. AB - BACKGROUND: Anesthetic drugs administered to immature animals may cause neurohistopathologic changes and alterations in behavior. The authors studied association between anesthetic exposure before age 4 yr and the development of reading, written language, and math learning disabilities (LD). METHODS: This was a population-based, retrospective birth cohort study. The educational and medical records of all children born to mothers residing in five townships of Olmsted County, Minnesota, from 1976 to 1982 and who remained in the community at 5 yr of age were reviewed to identify children with LD. Cox proportional hazards regression was used to calculate hazard ratios for anesthetic exposure as a predictor of LD, adjusting for gestational age at birth, sex, and birth weight. RESULTS: Of the 5,357 children in this cohort, 593 received general anesthesia before age 4 yr. Compared with those not receiving anesthesia (n = 4,764), a single exposure to anesthesia (n = 449) was not associated with an increased risk of LD (hazard ratio = 1.0; 95% confidence interval, 0.79-1.27). However, children receiving two anesthetics (n = 100) or three or more anesthetics (n = 44) were at increased risk for LD (hazard ratio = 1.59; 95% confidence interval, 1.06-2.37, and hazard ratio = 2.60; 95% confidence interval, 1.60-4.24, respectively). The risk for LD increased with longer cumulative duration of anesthesia exposure (expressed as a continuous variable) (P = 0.016). CONCLUSION: Exposure to anesthesia was a significant risk factor for the later development of LD in children receiving multiple, but not single anesthetics. These data cannot reveal whether anesthesia itself may contribute to LD or whether the need for anesthesia is a marker for other unidentified factors that contribute to LD. PMID- 19293701 TI - Critical thinking in anesthesia: eighth Honorary FAER Research Lecture. PMID- 19293702 TI - The pursuit of excellence: the 47th annual Rovenstine Lecture. PMID- 19293703 TI - Intermittent hypoxia and the practice of anesthesia. AB - Intermittent hypoxia, a powerful and unique stimulus, leads to physiologic changes that are distinct from those associated with either single or continuous hypoxic exposure. There is an accumulating body of evidence that the neurocognitive, inflammatory and cardiovascular symptoms that characterize the syndrome of obstructive sleep apnea are linked to the stimulus of intermittent hypoxia. In addition, altered sensitivities to opiates in children with obstructive sleep apnea have been linked to recurrent hypoxia during sleep. Therefore anesthesiologists should have an understanding of this important stimulus. PMID- 19293704 TI - A meta-analysis of clinical screening tests for obstructive sleep apnea. AB - The purpose of this meta-analysis is to compare clinical screening tests for obstructive sleep apnea and establish an evidence base for their preoperative use. Diagnostic odds ratios were used as summary measures of accuracy, and false negative rates were used as measures of missed diagnosis with each screening test in this review. Metaregression revealed that clinical models, logarithmic equations, combined techniques, cephalometry, and morphometry are significant characteristics, whereas body mass index, history of hypertension, and nocturnal choking are significant test elements associated with higher diagnostic accuracy. Test accuracy in repeated validation studies of the same screening test is variable, suggesting an underlying heterogeneity in either the clinical presentation of obstructive sleep apnea or the measured clinical elements of these models. Based on the false-negative rates, it is likely that most of the clinical screening tests will miss a significant proportion of patients with obstructive sleep apnea. PMID- 19293705 TI - Isoflurane differentially affects neurogenesis and long-term neurocognitive function in 60-day-old and 7-day-old rats. AB - BACKGROUND: Anesthetic agents cause cell death in the developing rodent brain and long-term, mostly hippocampal-dependent, neurocognitive dysfunction. However, a causal link between these findings has not been shown. Postnatal hippocampal neurogenesis affects hippocampal function into adulthood; therefore, the authors tested the hypothesis that isoflurane affects long-term neurocognitive function via an effect on dentate gyrus neurogenesis. METHODS: The S-phase marker 5 bromodeoxyuridine was administered at various times before, during, and after 4 h of isoflurane given to postnatal day (P)60 and P7 rats to assess dentate gyrus progenitor proliferation, early neuronal lineage selection, and long-term survival of new granule cell neurons. Fear conditioning and spatial reference memory was tested at various intervals from 2 weeks until 8 months after anesthesia. RESULTS: In P60 rats, isoflurane increased early neuronal differentiation as assessed by BrdU/NeuroD costaining, decreased progenitor proliferation for 1 day, and subsequently increased progenitor proliferation 5-10 days after anesthesia. In P7 rats, isoflurane did not induce neuronal lineage selection but decreased progenitor proliferation until at least 5 days after anesthesia. Isoflurane improved spatial reference memory of P60 rats long-term, but it caused a delayed-onset, progressive, persistent hippocampal deficit in P7 rats in fear conditioning and spatial reference memory tasks. CONCLUSION: The authors conclude that isoflurane differentially affects both neurogenesis and long-term neurocognitive function in P60 and P7 rats. Neurogenesis might mediate the long-term neurocognitive outcome after isoflurane at different ages. PMID- 19293707 TI - Use of the Pentax-AWS in 293 patients with difficult airways. AB - BACKGROUND: Several case reports have shown that the Pentax-AWS (Hoya Corporation, Tokyo, Japan), a new video laryngoscope, is useful in patients with difficult airways. METHODS: We assessed the effectiveness of the Pentax-AWS in two groups. Group 1 included 270 patients in whom direct laryngoscopy using a Macintosh laryngoscope had been difficult. Group 2 included 23 patients with predicted difficult intubation and difficult mask ventilation without previous use of the Macintosh laryngoscope. RESULTS: In group 1, the view of the glottis with the Macintosh laryngoscope was Cormack and Lehane grade 2 in 14 patients, grade 3 in 208 patients, and grade 4 in 48 patients. In 256 patients in whom the grade was 3 or 4 with the Macintosh laryngoscope, the view with the Pentax-AWS was either grade 1 or 2 in 255 patients (99.6%; 95% confidence intervals 97.8 100%). Tracheal intubation was successful with the Pentax-AWS in 268 of 270 patients (99.3%; 95% confidence interval 97.4-100%), and it failed (after two attempts) in two patients. In group 2, tracheal intubation was successful in 22 of 23 patients, and it failed in one patient. The reasons for failed intubation using the Pentax-AWS were failure to position the blade toward the glottic side of the epiglottis, inability to maneuver the endotracheal tube away from the arytenoids and into the trachea, and bleeding and swelling of the oropharynx. CONCLUSION: The success rate of tracheal intubation using the Pentax-AWS was high in patients with difficult laryngoscopy with a Macintosh laryngoscope and in patients with predicted difficult intubation. PMID- 19293708 TI - The effect of inhaled nitric oxide on the course of extracorporeal membrane oxygenation and the occurrence of hemorrhagic complications. AB - This study evaluated the relation between prior inhaled nitric oxide (iNO) and the time to initiation and duration of treatment with veno-arterial extracorporeal membrane oxygenation (ECMO) and the occurrence of hemorrhagic complications. A retrospective study was conducted in 59 human newborns treated for respiratory insufficiency with ECMO over a 5-year period. Patients received iNO before ECMO (iNO group) or not (control group). Both groups were compared for patient characteristics, postnatal age at the initiation of ECMO, duration of ECMO treatment, and hemorrhagic complications. There were no significant differences between the iNO group and the control group for patient characteristics and medication use before the ECMO treatment, except for norepinephrine. There was no significant difference for postnatal age at the initiation of ECMO and mean duration of ECMO treatment. We found no statistically significant difference in hemorrhagic complications between both groups. This persisted after correction for diagnosis and ECMO run-time in linear logistic regression analysis. Inhaled nitric oxide before ECMO did not result in a significant delay in the initiation of ECMO or longer duration of the ECMO. There was no significant relationship between the use of prior iNO and the occurrence of hemorrhagic complications during the ECMO treatment. PMID- 19293709 TI - Evaluation of nano-porous alumina membranes for hemodialysis application. AB - Globally, kidney failure has consistently been a major health problem. The number of patients suffering from kidney failure is radically increasing. Some studies forecast an exponential growth in the number of kidney failure patients during the coming years. This emphasizes the importance of hemodialysis (HD) membranes. Current dialysis membranes (cellulose based and synthetic polymer membranes) have irregular pore shapes and sizes, nonuniform pore distribution and limited reusable capability, which leads to low efficiency of toxin removal. New alumina membranes with uniform, controllable and well-structured nanoscale pores, channeled pores aligned perpendicular to the membrane plane, high porosity, high thermal and chemical resistance, and better mechanical properties are certainly preferable to currently used membranes. Determination of transport properties of alumina membranes will assist in the development of the alumina membranes for enhancing hemodialysis. Experiments were performed to evaluate hydraulic permeability, solute diffusive permeability, sieving coefficient, and clearance of four solutes (urea, creatinine, Vancomycin, and inulin) for alumina membrane. Based on comparison of these values against those of polyethersulfone (PES) membranes, transport performance of alumina membrane was determined. Hydraulic conductivity of the alumina membrane was approximately twice that of the PES membrane and inulin sieving coefficient for alumina membrane is approximately 21% higher than that for PES membrane. Alumina membrane has higher solute clearances and no albumin leakage, which makes it an effective replacement for current dialysis membranes. PMID- 19293710 TI - In vitro characterization and performance testing of the ension pediatric cardiopulmonary assist system. AB - In the last 40 years, mechanical circulatory support devices have become an effective option for the treatment of end-stage heart failure in adults. Few possibilities, however, are available for pediatric cardiopulmonary support. Ension Inc. (Pittsburgh, PA) is developing a pediatric cardiopulmonary assist system (pCAS) intended to address the limitations of existing devices used for this patient population. The pCAS device is an integrated unit containing an oxygenator and pump within a single casing, significantly reducing the size and blood-contacting surface area in comparison to current devices. Prototype pCAS devices produce appropriate flows and pressures while minimizing priming volume and preparation time. The pCAS was tested on a mock circulation designed to approximate the hemodynamic parameters of a small infant using a 10-Fr. extracorporeal membrane oxygenation inflow cannula and an 8-Fr. extracorporeal membrane oxygenation outflow cannula. Revision 4 of the device provided a flow rate of 0.42 L/min at 6,500 RPM. Revision 5, featuring improved impeller and diffuser designs, provided a flow rate of 0.57 L/min at 5,000 RPM. The performance tests indicate that for this cannulae combination, the pCAS pump is capable of delivering sufficient flows for patients <5 kg. PMID- 19293711 TI - Flow uniformity in oxygenators with different outlet port design. AB - This study reports on evaluation of the optimum design of a blood outlet port structure for providing uniform flow by visualizing the blood flow in an extracapillary membrane oxygenator. We tested a cylindrical type extracapillary membrane oxygenator, HPO-20. The HPO-20 has a tangential blood outlet port and is thus referred to as "Tangential HPO-20." We engineered "Vertical HPO-20" with a vertical blood outlet port by modifying the Tangential HPO-20. To visualize the blood-side flow, a total of 120 insulated copper-wire electrodes were placed in the "Tangential" and the Vertical HPO-20s. The test solution flow was visualized by the dimensionless fluid arrival time reaching each electrode. The test solution flow in the Tangential HPO-20 was not uniform, particularly at the outside blood channel. The flow was more uniform in the Vertical HPO-20. The blood flow in an extracapillary membrane oxygenator with a vertical blood outlet port is well distributed so that it produces more uniform blood flow than that with a tangential outlet port because of the small stagnation and reduced channeling. PMID- 19293712 TI - In-hospital outcomes of emergent and elective percutaneous coronary intervention in octogenarians. AB - OBJECTIVES: Although the prevalence of coronary artery disease increases with age, elderly patients are underrepresented in clinical trials. We sought to describe the short-term outcomes of octogenarians undergoing percutaneous coronary intervention (PCI) in both emergent and elective settings. METHODS: We retrospectively identified all octogenarians undergoing PCI at our institution from January 2002 to December 2005. The outcomes of those undergoing primary PCI for ST-segment elevation myocardial infarction (group A) were compared with a randomly selected cohort of octogenarians undergoing elective PCI (group B). RESULTS: Group A comprised 91 patients and group B 88 patients. Procedure success was high with Thrombolysis in Myocardial Infarction grade 3 flow achieved in greater than 90% of patients in both groups. Group A experienced more postprocedure morbidity including acute renal failure (47 vs. 8%) and bleeding requiring transfusion (51 vs. 17%) (P<0.0001 for both comparisons). In-hospital mortality was higher in group A (21 vs. 0%). In multivariate analysis, the risk of in-hospital mortality was predicted by lower admission systolic blood pressure (odds ratio 1.026 per point decrease in systolic blood pressure; 95% confidence interval 1.003-1.049; P = 0.030) and development of cardiogenic shock (odds ratio 7.506; 95% confidence interval, 1.865-30.207; P = 0.005). Mortality in group A was significantly higher among patients with cardiogenic shock (42 vs. 6%, P<0.001). CONCLUSION: PCI is a safe and highly effective procedure in octogenarians during both emergent and elective settings. However, elderly patients undergoing primary angioplasty who develop hemodynamic instability during ST-segment elevation myocardial infarction remain at increased risk for mortality. Measures to reduce postprocedure bleeding and renal dysfunction may improve outcomes among elderly patients undergoing PCI. PMID- 19293713 TI - B-type natriuretic peptide: a strong predictor of early and late mortality in patients with acute chest pain without ST-segment elevation in the emergency department. AB - BACKGROUND: The prognostic importance of early measurement of B-type natriuretic peptide (BNP) in patients with acute chest pain while the diagnosis is still uncertain is unknown. We determined the prognostic value of BNP in these patients immediately after presenting to the emergency department. METHODS: Seven hundred and twenty-three consecutive individuals with suspicious ischemic acute chest pain and no ST-segment elevation were prospectively evaluated using a systematic diagnostic strategy and followed for 1 year. Acute coronary syndrome was diagnosed in 326 patients during their hospital stay. RESULTS: In the follow-up, 15 (2.1%) patients of the whole cohort died of cardiac cause at 1 month and 51 (7.1%) at 1 year. Patients who died had significantly higher admission BNP levels than survivors and this correlation proved linear according to quartile levels. Patients with BNP greater than 101 pg/ml had 13 times higher rate of 1-month mortality (P<0.0001) and 5.3 times higher rate of 1-year mortality (P<0.0001) than patients with BNP of 101 pg/ml or less. Multiple logistic regression analysis disclosed BNP as a strong independent predictor of 1-month and 1-year mortality adding significant prognostic information over traditional risk markers. CONCLUSION: Admission BNP is an independent and powerful marker of early and late cardiac mortality in patients with acute chest pain without ST-segment elevation. These results suggest that BNP should be measured upon arrival at the emergency department for risk stratification in all these patients. PMID- 19293714 TI - The challenges in developing a rational cannabis policy. AB - PURPOSE OF REVIEW: A rational cannabis policy would arguably be one that minimized the harms of both cannabis use and the legal policies adopted to control its use. We, therefore, review recent epidemiological evidence on the harmful effects of cannabis use and social research on the costs and benefits of cannabis prohibition. RECENT FINDINGS: Epidemiological evidence suggests that cannabis increases the risk of road crash injury if users drive while intoxicated. When used chronically, cannabis can produce dependence, respiratory disease and psychotic symptoms, especially in vulnerable young adults. It probably also increases poor educational outcomes and possibly increases the use of other illicit drugs, although it is debated whether these relationships are causal. Proponents of a relaxation of cannabis prohibition argue that prohibition has failed to deter cannabis use, incurs substantial economic costs, has generated a large black market, has increased the potency of cannabis and users' access to other drugs and involves foregone tax revenue from the legal sale of cannabis. SUMMARY: Development of a more rational cannabis policy requires better evaluations of both the health consequences of regular cannabis use and of the costs and benefits of enforcing the existing prohibition on its use. It also requires the liberalization of the international control system to allow member states to experiment with different methods of regulating and controlling cannabis use. PMID- 19293715 TI - Neurobiology of human affiliative behaviour: implications for psychiatric disorders. AB - PURPOSE OF REVIEW: Most of our previous knowledge about neurobiology of affiliation is based on animal research. However, during the last couple of years, there has been a surge of research investigating the neurobiological underpinnings of human affiliative behaviour. This article aims to review recent brain imaging and neuropeptide studies that investigated human affiliation. RECENT FINDINGS: Findings from brain-imaging studies have increased our knowledge of the neural networks critical for sensitivity to social reward, social cooperation and empathy. Some of the inter-individual and between-sex differences in personality and temperament appear to be driven by structural and functional variations in the brain regions processing social rewards. Furthermore, oxytocin exerts its effects by modulating affiliation-related neural networks. Impairments in affiliation-related brain networks and neuropeptides seem to be potentially important for pathophysiology of a range of mental disorders such as autism, schizophrenia and psychopathy. SUMMARY: Human research investigating the neurobiology of affiliation has potentially important implications for the understanding and management of mental disorders. One example is the potential role of oxytocin as a therapeutic agent; however, these preliminary findings need to be rigorously tested with further studies. Multimethod studies that evaluate brain imaging, genetics and neurochemistry within the same sample will be important to further advance our understanding of human affiliation. PMID- 19293716 TI - Correlates of exercise among coronary heart disease patients: review, implications and future directions. AB - Despite the well-documented benefits of exercise, adherence among patients with coronary heart disease (CHD) has been low during and after cardiac rehabilitation (CR) as well as among patients not attending CR. Therefore, an understanding of the factors that influence exercise in this population is crucial to assist in the development of effective interventions. The goal of this review was to document the correlates of exercise of CHD patients in all CR contexts from a social-ecological perspective that addresses multiple levels of influence on exercise. The search strategy included PubMed, PsychINFO and Web of Science databases. In all, 121 studies, examining 32 different correlates of exercise, with a total of 25 217 participants were included. Across all CR contexts, six variables were consistently related to exercise (self-regulatory self-efficacy, health status, intention, perceived control, beliefs/benefits and previous physical activity). Several variables were also related to exercise in three of four contexts (e.g. task self-efficacy, perceived barriers, attitude, action planning, sex and employment status). Many of the variables consistently related to exercise may be amenable to change through the development and implementation of appropriate interventions. Specific suggestions are made for each relevant variable to assist CR staff and other healthcare practitioners in 32 promoting exercise among CHD patients. Current gaps in literature such as a lack of prospective studies and research examining broader (e.g. policy level) correlates are also discussed. PMID- 19293717 TI - Glycemia at admission: the metabolic echocardiography in acute coronary syndrome patients. AB - BACKGROUND: Ventricular dysfunction in acute coronary syndrome (ACS) patients is a recognized predictor of in-hospital and post-discharge morbidity and mortality. Recently, hyperglycemia at admission has been considered as an important marker of poor in-hospital prognosis. AIM: To characterize an ACS population and to identify independent predictors of one-year mortality. METHODS: This study included 1179 consecutive patients admitted to a single coronary care unit for acute coronary syndrome between May 2004 and December 2006. Patients were followed up for 12 months after ACS. RESULTS: In our population, 70.9% of patients were male, with a mean age of 66.6+/-12.5 years; 15.7% were admitted by unstable angina, 44.7% by non-ST elevation myocardial infarction and 38.5% by ST elevation myocardial infarction; 16% of patients had left ventricular systolic dysfunction during the index admission. In-hospital mortality was 3.5% and complications occurred in 5.6% of patients. Mortality rate at 1-year of follow-up was 8.8% and rehospitalization rate for heart failure was 5.5%. After multivariate regression analysis, left ventricular systolic dysfunction [odds ratio (OR): 3.58; confidence interval (CI): 1.57-8.16], glycemia at admission >137 mg/dl (OR: 3.58; CI: 1.52-8.47) and age >65 years (OR: 2.67; CI: 1.08-6.59) were independent predictors of 1-year mortality. CONCLUSION: In this population, hyperglycemia at admission was an independent predictor of 1-year mortality, with a predictive value equivalent to that of left ventricular systolic dysfunction. This fact, never before described, emphasizes the importance of metabolic abnormalities and their control in the prognosis of ACS patients. PMID- 19293718 TI - Results achieved by emergency physicians in teaching basic cardiopulmonary resuscitation to secondary school students. AB - OBJECTIVE: We investigated the results obtained with a basic cardiopulmonary resuscitation (b-CPR) program (PROCES) specifically designed for secondary school students (14-16 years old) and taught by emergency physicians. METHODS: We used a multiple-choice test with 20 questions (10 on theory and 10 on skills) answered before and immediately after and 1 year after receiving the b-CPR course. Satisfactory learning was considered when at least 8 out of 10 skill questions were correctly answered. We investigated student variables associated with better immediate and deferred (1 year after) PROCES performance. We compared the results with those obtained using a more standardized program to teach b-CPR to police cadets. RESULTS: We enrolled 600 high school students. PROCES achieved significant improvement in overall, theory and skill marks immediately after the course (P<0.001), with a significant decay in all of them 1 year after the course (P<0.001). Satisfactory learning was achieved by 57% of school students immediately after PROCES and by 37% when assessed 1 year later. Students without pending study subjects (P=0.001) and those from private schools (P<0.01) achieved significantly better performance immediately after PROCES and only female students achieved greater performance 1 year after the course (P<0.05). With respect to police cadets instructed through a standardized course, immediate satisfactory learning of school students was lower (79 vs. 57%, respectively; P<0.001), whereas deferred satisfactory learning was higher (23 vs. 37%, respectively; P<0.05). CONCLUSION: Emergency physicians can satisfactorily instruct secondary school students in b-CPR using PROCES, and this specific program achieves a reasonable amount of satisfactory learning. PMID- 19293719 TI - Clinical meaning of pepsinogen test and Helicobacter pylori serology in the health check-up population in Korea. AB - OBJECTIVE: This study was performed to assess the affects of age, sex, and Helicobacter pylori status on pepsinogen testing for atrophic gastritis and to establish the clinical implications of pepsinogen test results and H. pylori serology in a Korean population presenting for a health check-up. METHODS: Serum pepsinogen (PG) I and PG II, and H. pylori IgG were measured in 1485 adults. The PG values were analyzed based on age, sex, and H. pylori status, and the cutoff value for atrophic gastritis was determined. RESULTS: Serum PG I (sPGI) and sPGII were higher in H. pylori positive than in H. pylori negative individuals (sPGI, 56.3 vs. 42.2 microg/l, P<0.001; sPGII, 17.5 vs. 8.0 microg/l, P<0.001). The PG I/II ratio was lower in H. pylori positive than in H. pylori negative individuals (3.7 vs. 6.0, P<0.001). The sPGII and PG I/II ratio had a positive (r=0.132, P<0.001) and negative correlation with age (r=-0.229, P<0.001), respectively. Men had a higher sPGI (54.7 microg/l) than did women (48.4 microg/l) (P<0.001) but the PG I/II ratio was not statistically different and neither the atrophic gastritis. The PG I/II cutoff value for atrophic gastritis was 6.0 for H. pylori negative and 3.0 for H. pylori positive individuals. sPGI and sPGII were, however, not specific for atrophic gastritis. CONCLUSION: The H. pylori IgG status, age, and sex were associated with the serum PG levels. To increase the efficacy of the PG I/II ratio for the detection of atrophic gastritis, the cutoff value of the PG I/II ratio should be stratified according to the H. pylori IgG status in the Korean population presenting for a health check-up. PMID- 19293720 TI - Variceal bleeding in primary biliary cirrhosis patients: a subgroup with improved prognosis and a model to predict survival after first bleeding. AB - BACKGROUND: Varices are a late complication in primary biliary cirrhosis (PBC). However, patients without clinical jaundice do bleed from varices; whether their prognosis differs is unknown. AIM: Evaluate PBC patients, particularly those with bilirubin 34 micromol/l (P=0.001, log rank test). Hazard ratios (95% confidence intervals) for independent predictors of mortality after bleeding were: age 1.02 (1-1.05), log10 bilirubin 4.64 (2.56-8.41), ascites 2.13 (1.29-3.51) and hepatic encephalopathy 2.72 (1.56-4.74). CONCLUSION: Variceal bleeding complicates histologically advanced PBC. A distinct subgroup with near normal bilirubin and lower alkaline phosphatase first presents with variceal bleeding in 50% of cases and has a better prognosis than jaundiced PBC variceal bleeders. PMID- 19293721 TI - Assessment of splanchnic blood flow using magnetic resonance imaging. AB - BACKGROUND AND AIMS: The splanchnic circulation has an important function in the body under both physiological and pathophysiological conditions. Despite its importance, no reliable noninvasive procedures for estimating splanchnic circulation have been established. The aim of this study was to evaluate MRI as a tool for assessing intra-abdominal blood flows of the aorta, portal vein (VPO) and the major intestinal and hepatic vessels. METHODS: In nine healthy volunteers, the proximal aorta (AOP) and distal abdominal aorta (AOD), superior mesenteric artery (SAM), celiac trunk (CTR), hepatic arteries (common and proper hepatic arteries, AHC and AHP, respectively), and VPO were localized on contrast enhanced magnetic resonance angiography images. Volumetric flow was measured using a two-dimensional cine echocardiogram-gated phase contrast technique. Measurements were taken before and 30 min after continuous intravenous infusion of somatostatin (250 microg/h) and were independently evaluated by two investigators. RESULTS: Blood flow measured by MRI in the VPO, SAM, AOP, AHP, and CTR significantly decreased after drug infusion. Flows in the AOD and AHC showed a tendency to decrease (P>0.05). Interrater agreement on flows in MRI was very good for large vessels (VPO, AOP, and AOD), with a concordance correlation coefficient of 0.94, as well as for smaller vessels such as the CTR, AHC, AHP, and SAM (concordance correlation coefficient =0.78). CONCLUSION: Somatostatin induced blood flow changes in the splanchnic region were reliably detected by MRI. MRI may be useful for the noninvasive assessment of blood flow changes in the splanchnic region. PMID- 19293722 TI - Systematic review on drug and diet-induced endoscopic remission in Crohn's disease. AB - BACKGROUND: Neither the rate of endoscopic remission (ER) in Crohn's disease (CD) after therapy nor its role in patients' prognosis is well defined. AIM: To systematically review the current evidence on the proportion of ER of different therapies in patients with Crohn's disease and its relation with clinical outcomes. METHODS: Databases (MEDLINE and Cochrane) and manual search of manuscripts found 482 titles. Data was extracted from 24 manuscripts. RESULTS: Ten different methods were used to assess endoscopic outcomes. Corticosteroids induced a pooled proportion of patients with no ulcerations at endoscopic follow up of 17% (95% confidence interval: 12-22%) lower than that found with infliximab [44% (34-53%)], diet [43% (33-52%)] or azathioprine [54% (38-69%)] (P<0.0001). Enteric diets and infliximab were associated with 61% (52-70%) and 70% (62-78%) reduction in endoscopic scores, respectively, significantly higher than corticosteroids [45% (39-52%)] (P=0.01) and placebo [12% (1-22%)] (P<0.0001). A linear relation between ER and clinical remission was observed with infliximab (r=0.931). Only one study tried to assess the direct correlation between ER and patients' prognosis. CONCLUSION: Available treatments induce significant endoscopic improvement. However, pooled results should be cautiously interpreted because of the diversity of measurements. A better definition of endoscopic outcomes and a prospective validation of their relevance in patients with Crohn's disease are needed. PMID- 19293723 TI - The geriatric hand: correlation of hand-muscle function and activity restriction in elderly. AB - On the basis of the importance of hand manipulation in activities of daily living (ADL), deterioration of hand function because of various factors reduces quality and independence of life of the geriatric population. The aim of this study was to identify age-induced changes in manual function and to quantify the correlations between hand-muscle function and activity restriction in the geriatric age group, through grip and pinch measurements and a set of questionnaires. Twenty-four geriatric (aged 65-79 years) volunteers participated in the study. Bilateral grip and pinch strengths have been recorded. To document impairment of manual functions, self-estimated hand function, Duruoz and Dreiser hand indices, Geriatrics-Arthritis Impact Measurement Scale (GERI-AIMS) manual dexterity questionnaires have been completed. Activity restriction and quality of life of these patients were inquired with short form (SF)-36 and Instrumental Activities of Daily Living (IADL) scores. Grip and pinch strengths correlated best with Duruoz and Dreiser indices. Similarly, SF-36 and IADL had higher correlation coefficients for Duruoz and Dreiser indices. A very good correlation between IADL and SF-36 was calculated too. Male and female participants revealed statistically significant differences for grip and pinch strengths as well as self-estimated hand function and SF-36. Another result was that none of our parameters, including grip strength and SF-36 had differed significantly between the 65-70 and 70-79 years age subgroups. However, grip strength displayed statistically significant lower values when compared with young adult mean values of a previous study. Our data in this study support the hypothesis that hand muscle function correlates with functional dependency in the elderly. Manual function can be determined by grip strength in addition to multiple available functional tools. In this study, Dreiser and Duruoz hand function indices were the best to correlate with ADL and quality of life. PMID- 19293724 TI - The myocardial infarction associated CDKN2A/CDKN2B locus on chromosome 9p21 is associated with stroke independently of coronary events in patients with hypertension. AB - OBJECTIVE: We tested whether two single-nucleotide polymorphisms (SNPs) (rs2383207 and rs10757278), previously strongly associated with myocardial infarction, are independently associated with stroke and coronary events in patients with hypertension. METHODS: The Nordic Diltiazem study compared the effects of calcium antagonist and beta-blocker or diuretic-based antihypertensive treatment on cardiovascular events in 10 881 patients with hypertension, of whom 5262 patients provided DNA for the present study. We related SNPs rs2383207 and rs10757278 to stroke and to myocardial infarction and coronary revascularizations (coronary events) using crude and multivariate adjusted Cox proportional hazards models. RESULTS: The G-allele of both SNPs predicted coronary events in crude recessive models [hazard ratios = 1.36, 95% confidence interval (CI) = 1.04-1.79, P = 0.02 for rs10757278 and hazard ratios = 1.40, 95% CI = 1.08-1.81, P = 0.01 for rs2383207] as well as after adjustment for classical cardiovascular risk factors. The G-allele of both SNPs predicted incident stroke in crude additive models [rs2383207 hazard ratios = 1.25 (95% CI = 1.02-1.53), P = 0.04 and rs10757278 hazard ratios = 1.34 (95% CI = 1.09-1.65), P = 0.006], as well as after adjustment for classical cardiovascular risk factors and after additional adjustment for prevalent and incident coronary events, atrial fibrillation, ischemic heart disease and congestive heart failure. As was the case for coronary events, the excess genetic risk of stroke was driven by subjects homozygous for the risk allele. CONCLUSION: Genetic variation at the CDKN2A/CDKN2B locus predicts stroke in hypertensive patients. The genetic association with stroke is independent of classical cardiovascular risk factors and of all prevalent and incident coronary events, suggesting that gene variation at this locus promotes either atherosclerosis or another disease mechanism that is common to both coronary and cerebrovascular disease. PMID- 19293725 TI - Differences in healthy lifestyle characteristics between adults with prehypertension and normal blood pressure. AB - BACKGROUND: Identifying differences in modifiable lifestyle factors between persons with prehypertension and normal blood pressure (BP) can help improve prevention efforts. METHODS: Data from the 1999-2004 National Health and Nutrition Examination Survey were analyzed (in 2008) for persons aged at least 20 years (n = 11 194). Differences in five healthy lifestyle characteristics were examined by BP status (normal, prehypertension, and hypertension). Additionally, differences in lifestyle characteristics by sex, race/ethnicity, and education among those with prehypertension were analyzed. RESULTS: Overall, 32.8% of adults had a normal BMI, 75.3% did not smoke, 31.3% were regularly physically active, 57.7% were moderate drinkers or nondrinkers, and 28.1% had a low sodium intake; only 2% had all five characteristics. Almost 40% had a normal BP and 30.3% were prehypertensive. Those with prehypertension were less likely to have a normal BMI than normotensive individuals [adjusted odds ratio (OR) = 0.63, 95% confidence interval (CI) = 0.56-0.70], to be regularly active (adjusted OR = 0.85, 95% CI = 0.74-0.98), and to moderately/not drink (adjusted OR = 0.88, 95% CI = 0.80-0.97). Those with prehypertension or hypertension were less likely to have at least three or four healthy lifestyle characteristics compared with those with normal BP. Among 3168 persons with prehypertension, some sex, race/ethnic, and education level differences in the prevalence of healthy lifestyles were observed. CONCLUSION: Differences in healthy lifestyle factors were observed by BP status, but the prevalence of healthy lifestyle factors is suboptimal among the population as a whole. PMID- 19293726 TI - Effects of antihypertensive treatment on small artery remodelling. AB - Although it is recognized that the cause of hypertension can be various, once blood pressure has become established structural changes emerge in the systemic vasculature. In medium- and large-sized vessels, as in the left ventricle, there is clear histological evidence of hypertrophy of the medial smooth muscle layers but, downstream in small arteries, which modulate vascular resistance, other changes occur. In essential hypertension, the smooth muscle cells of small vessels are restructured around a smaller lumen, but there is no evidence of hypertrophy or hyperplasia of the vascular wall. In secondary forms of hypertension, which tend to be representative of severer forms of the disease, hypertrophic remodelling is observed. Similarly, in non-insulin-dependent diabetes mellitus, irrespective of whether blood pressure accompanies this disorder or not, hypertrophy is also seen. The presence of architectural alterations in the vascular wall of small arteries may have a strong prognostic significance in patients, and this may be over and above all other known cardiovascular risk factors. Although it is yet to be established whether regression of such changes should be a goal of effective antihypertensive therapy, there is a body of evidence emerging indicating that different classes of antihypertensive drug have a varied effect on reversing vascular structure both in humans and animal models of genetic and experimental hypertension. However, at present, there are no data available concerning the prognostic impact of regressing vascular structural alterations in hypertension, and this must be an urgent research priority. PMID- 19293727 TI - Association between insulin, leptin, adiponectin and blood pressure in youth. AB - OBJECTIVE: To examine whether insulin, leptin and adiponectin are independent correlates of blood pressure (BP) in a large population-based sample of children and adolescents. METHODS: We studied 655 boys and 667 girls aged 9, 13 and 16 years who participated in the Quebec Child and Adolescent Health and Social Survey, a province-wide school-based survey conducted in 1999. RESULTS: Strong, positive univariate associations between BMI, insulin and leptin Z-scores, and both systolic and diastolic BP were found in both sexes. Adiponectin Z-scores were negatively associated with systolic BP in girls only. In multivariate analyses only BMI and insulin Z-scores were significantly associated with systolic BP. In boys, each 1 SD increase in BMI was associated with a 4 mmHg increase in mean systolic BP; each 1 SD increase in insulin was associated with a 1 mmHg increase in mean systolic BP. Only insulin Z-scores were independently associated with diastolic BP in both sexes. For each 1 SD increase in insulin, there was a 1 mmHg increase in mean diastolic BP in boys. Similar to systolic BP, the magnitude of the effect of insulin Z-scores on diastolic BP increased as a function of BMI Z-scores in girls. CONCLUSION: Independently of BMI, insulin is a strong correlate of systolic and diastolic BP in youth. Although animal studies support a role for leptin and adiponectin in controlling BP, they are not independently associated with BP in youth. PMID- 19293729 TI - Moxibustion for treating menopausal hot flashes: a randomized clinical trial. AB - OBJECTIVE: A hot flash is a general postmenopausal symptom experienced by approximately 75% of climacteric women. Women often turn to complementary and alternative medicines to relieve hot flashes. Moxibustion is one such medication. The aim of this study was to evaluate the effect of moxibustion on hot flashes in perimenopausal and postmenopausal women. METHODS: Fifty-one participants were randomly assigned into three groups, namely, moxibustion 1, moxibustion 2, and waiting list (control). The moxibustion groups received 14 sessions of moxibustion treatment for 4 weeks at acupuncture points. Our protocol was supported by evidence from clinical experts (Moxa 1) or published literature (Moxa 2), and we followed all participants for an additional 2 weeks after the end of the study. Our primary outcome measures were frequency and severity of hot flashes. Secondary outcome parameters included quality of life (Menopausal Specific Quality of Life Scale) and Menopause Rating Scale. RESULTS: Fifty-one women participated in our study. By week 4, the difference in severity and frequency of hot flashes had become statistically significant between the treatment groups and the control participants. In addition, there was a statistically significant difference in Menopausal-Specific Quality of Life Scale scores between treatment group 2 and the other groups. CONCLUSIONS: Our results suggest that moxibustion reduces both the frequency and severity of menopausal hot flashes as compared with those in control participants. We would recommend further studies with larger samples and possibly including placebo controls. PMID- 19293728 TI - D1-like receptors inhibit insulin-induced vascular smooth muscle cell proliferation via down-regulation of insulin receptor expression. AB - OBJECTIVE: Vascular smooth muscle cell (VSMC) proliferation is central to the development of vascular diseases, including hypertension, which is regulated by numerous hormones and humoral factors. Our previous study showed that the stimulatory effect of norepinephrine on VSMC proliferation is inhibited by D1 like receptors and the D3 dopamine receptor, a member of the D2-like receptor family. Insulin is a proliferative hormone but it is not known if there is any interaction between insulin and D1-like receptors. We hypothesized that Dl-like receptors may have an inhibitory effect on the insulin-induced VSMC proliferation; aberrant insulin and Dl-like receptor functions could be involved in the pathogenesis of essential hypertension. METHODS: VSMC proliferation was determined by [H]-thymidine incorporation; insulin receptor mRNA and protein expressions were determined by RT-PCR, immunoblotting, and immunohistochemistry. RESULTS: Insulin increased VSMC proliferation in immortalized aortic A10 cells, determined by [H]-thymidine incorporation. Although the D1-like receptor, by itself, had no effect on VSMC proliferation, stimulation with fenoldopam, a D1 like receptor agonist, inhibited the stimulatory effect of insulin. The inhibitory effect of fenoldopam on insulin-mediated VSMC proliferation was receptor specific, because its effect could be blocked by SCH23390, a D1-like receptor antagonist. Fenoldopam also inhibited insulin receptor mRNA and protein expression, which was time dependent and concentration dependent. A PKC or MAP kinase inhibitor blocked the inhibitory effect of fenoldopam on insulin receptor expression, indicating that PKC and MAP kinase were involved in the signaling pathway. CONCLUSION: The inhibitory effect of D1-like receptors on insulin mediated VSMC proliferation may play an important role in the regulation of blood pressure. PMID- 19293730 TI - Loss feedback negativity elicited by single- versus conjoined-feature stimuli. AB - Event-related brain potential studies show that negative feedback in guessing tasks elicits a medial frontal negativity. Most theory and experimentation concerning this feedback-related negativity (FRN) has assumed that the FRN has little relationship to the perceptual characteristics of the feedback. This study challenges this assumption. We used a single visual feature or a conjunction of features to indicate reward feedback in a gambling task. In the single-feature condition, losses elicited a larger FRN than gains; in the conjoined-feature condition, that difference was not observed. The results are consistent with the proposal that the FRN is modulated by the deviation of feedback stimuli from a perceptual template. Future studies must not confound the perceptual properties and the valence of reward feedback. PMID- 19293731 TI - Goldfish CB1 mRNA expression is affected by fasting and anandamide administration. AB - The endocannabinoid system has a well-documented pivotal role in the control of mammalian feeding response; nevertheless, some evidence is available regarding a similar role in nonmammalian vertebrates and invertebrates. As in the bonyfish Carassius auratus, CB1 cannabinoid receptors are abundant in brain regions involved in the control of food intake, and fasting affects endocannabinoid levels, in this study the effects of food deprivation and anandamide administration on CB1 expression were evaluated. Fasting led to a time-dependent increase of CB1 mRNA levels in the forebrain, an effect reversed by refeeding. Furthermore, the administration of exogenous anandamide reduced CB1 expression in food-deprived goldfish. Our results support the involvement of CB1 receptors in the control of energy intake in nonmammalian vertebrates. PMID- 19293732 TI - Attack on the Taj Mahal Palace Hotel: a proxy war on America? PMID- 19293734 TI - Emergency department preparedness for radiation emergencies in the Philadelphia area. PMID- 19293735 TI - From the editor-in-chief. PMID- 19293736 TI - Developing consensus on appropriate standards of hospital disaster care: ensuring that the needs of children are addressed. PMID- 19293737 TI - Responding to gender-based violence in disasters: grappling with research methods to clear the way for planning. PMID- 19293738 TI - World update: cholera crisis in Zimbabwe. PMID- 19293739 TI - Thermal stress and point-of-care testing performance: suitability of glucose test strips and blood gas cartridges for disaster response. AB - OBJECTIVE: Point-of-care testing (POCT) devices are deployed in the field for emergency on-site testing under a wide range of environmental conditions. Our objective was to evaluate the performance of glucose meter test strips and handheld blood gas analyzer cartridges following thermal stresses that simulate field conditions. METHODS: We evaluated electrochemical and spectrophotometric glucose meter systems and a handheld blood gas analyzer. Glucose test strips were cold-stressed (-21 degrees C) and heat-stressed (40 degrees C) for up to 4 weeks. Blood gas cartridges were stressed at -21 degrees C, 2 degrees C, and 40 degrees C for up to 72 hours. Test strip and cartridge performance was evaluated using aqueous quality control solutions. Results were compared with those obtained with unstressed POCT strips and cartridges. RESULTS: Heated glucose test strips and blood gas cartridges yielded elevated results. Frozen test strips and cooled cartridges yielded depressed glucose and blood gas results, respectively. Frozen cartridges failed. CONCLUSIONS: The performance of glucose test strips and blood gas cartridges was affected adversely by thermal stresses. Heating generated elevated results, and cooling depressed results. Disaster medical assistance teams and emergency medical responders should be aware of these risks. Field POCT devices must be robust to withstand adverse conditions. We recommend that industry produce POCT devices and reagents suitable for disaster medical assistance teams. PMID- 19293740 TI - Increased gender-based violence among women internally displaced in Mississippi 2 years post-Hurricane Katrina. AB - OBJECTIVES: Although different types of gender-based violence (GBV) have been documented in disaster-affected populations, no studies have documented a quantitative increase in rates of GBV among populations living in protracted displacement after a disaster. We aimed to assess the change in rates of GBV after Hurricane Katrina among internally displaced people (IDPs) living in travel trailer parks in Mississippi. METHODS: The study design included successive cross sectional randomized surveys, conducted in 2006 and 2007, among IDPs in Mississippi using a structured questionnaire. We sampled 50 travel trailer parks in 9 counties in Mississippi in 2006, and 69 parks in 20 counties in 2007. A total of 420 female respondents comprised the final sample. We measured respondent demographics, forms of GBV including sexual and physical violence further subtyped by perpetrator, suicidal ideation, suicide attempt, and Patient Health Questionnaire-9-assessed depression. RESULTS: Respondents had a mean age of 42.7 years. The crude rate of new cases of GBV among women increased from 4.6/100,000 per day to 16.3/100,000 per day in 2006, and remained elevated at 10.1/100,000 per day in 2007. The increase was primarily driven by the increase in intimate partner violence. GBV experience was significantly associated with increased risk for poor mental health outcomes. CONCLUSIONS: Overall, the rate of GBV, particularly intimate partner violence, increased within the year following Hurricane Katrina and did not return to baseline during the protracted phase of displacement. Disaster planning efforts should incorporate plans to decrease the incidence of GBV following a disaster, and to ensure adequate services to people with postdisaster GBV experience. PMID- 19293741 TI - Developing consensus on appropriate standards of disaster care for children. AB - BACKGROUND: Neither professional consensus nor evidence exists to guide the choice of essential hospital disaster interventions. The objective of our study was to demonstrate a method for developing consensus on hospital disaster interventions that should be regarded as essential, quantitatively balancing needs and resources. METHODS: A panel of pediatric acute care practitioners developed consensus using a modified Delphi process. Interventions were chosen such that workload per staff member would not exceed the previously validated maximum according to the Therapeutic Intervention Scoring System. Based on published models, it was assumed that the usual numbers of staff would care for a disaster surge of 4 times the usual number of intensive care and non-intensive care hospital patients. RESULTS: Using a single set of assumptions on constrained resources and overwhelming needs, the panel ranked and agreed on essential interventions. A number of standard interventions would exceed crisis workload constraints, including detailed recording of vital signs and fluid balance, administration of vasoactive agents, invasive monitoring of pressures (central venous, intraarterial, intracranial), dialysis, and tube feedings. CONCLUSIONS: The quantitative methodology and consensus development process described in the present report may have utility in future planning. Groups with appropriate expertise must develop action plans according to authority within each jurisdiction, addressing likely disaster scenarios, according to the needs in each medical service region, using available regional resources, and accounting for the capabilities of each institution. PMID- 19293742 TI - Variations in disaster preparedness by mental health, perceived general health, and disability status. AB - OBJECTIVES: Chronic medical and mental illness and disability increase vulnerability to disasters. National efforts have focused on preparing people with disabilities, and studies find them to be increasingly prepared, but less is known about people with chronic mental and medical illnesses. We examined the relation between health status (mental health, perceived general health, and disability) and disaster preparedness (home disaster supplies and family communication plan). METHODS: A random-digit-dial telephone survey of the Los Angeles County population was conducted October 2004 to January 2005 in 6 languages. Separate multivariate regressions modeled determinants of disaster preparedness, adjusting for sociodemographic covariates then sociodemographic variables and health status variables. RESULTS: Only 40.7% of people who rated their health as fair/poor have disaster supplies compared with 53.1% of those who rate their health as excellent (P < 0.001). Only 34.8% of people who rated their health as fair/poor have an emergency plan compared with 44.8% of those who rate their health as excellent (P < 0.01). Only 29.5% of people who have a serious mental illness have disaster supplies compared with 49.2% of those who do not have a serious mental illness (P < 0.001). People with fair/poor health remained less likely to have disaster supplies (adjusted odds ratio [AOR] 0.69, 95% confidence interval [CI] 0.50-0.96) and less likely to have an emergency plan (AOR 0.68, 95% CI 0.51-0.92) compared with those who rate their health as excellent, after adjusting for the sociodemographic covariates. People with serious mental illness remained less likely to have disaster supplies after adjusting for the sociodemographic covariates (AOR 0.67, 95% CI 0.48-0.93). Disability status was not associated with lower rates of disaster supplies or emergency communication plans in bivariate or multivariate analyses. Finally, adjusting for the sociodemographic and other health variables, people with fair/poor health remained less likely to have an emergency plan (AOR 0.66, 95% CI 0.48-0.92) and people with serious mental illness remained less likely to have disaster supplies (AOR 0.67, 95% CI 0.47-0.95). CONCLUSIONS: People who report fair/poor general health and probable serious mental illness are less likely to report household disaster preparedness and an emergency communication plan. Our results could add to our understanding of why people with preexisting health problems suffer disproportionately from disasters. Public health may consider collaborating with community partners and health services providers to improve preparedness among people with chronic illness and people who are mentally ill. PMID- 19293743 TI - Willingness of health care personnel to work in a disaster: an integrative review of the literature. AB - Effective hospital surge response in disaster depends largely on an adequate number of personnel to provide care. Studies appearing since 1991 indicate health care personnel may not be willing to work in all disaster situations-and if so, this could degrade surge response. A systematic review of the literature was conducted to determine the state of the evidence concerning the willingness of health care personnel to work in disaster. The aims of this review are to collate and assess the literature concerning willingness of health care personnel to work during a disaster, to identify gaps in the literature as areas for future investigation, and to facilitate evidence-based disaster planning. Twenty-seven studies met inclusion criteria (25 quantitative and 2 qualitative studies). The current evidence indicates there may be certain factors related to willingness to work (or lack of willingness) in disaster including the type of disaster, concern for family, and concerns about personal safety. Barriers to willingness to work have been identified including pet care needs and the lack of personal protective equipment. This review describes the state of an emerging area of science. These findings have significant implications for community and organizational emergency planning and policymaking in an environment defined by limited resources. PMID- 19293744 TI - Active steps to promote influenza vaccination. PMID- 19293752 TI - Influence of preservation method on histopathological lesions of kidney allografts. AB - BACKGROUND: Cold storage (CS) of cadaveric kidneys procured from hemodynamically stable donors for less than 24 hours is a safe procedure. Some papers indicate that continuous pulsatile machine perfusion (MP) allows for extension of preservation times, permits a wider use of kidneys damaged by preagonal ischemia, results in superior immediate function rate as compared to CS and improves long term graft survival. The aim of the study was to evaluate the influence of kidney preservation method prior to transplantation on the characteristics of histopathological lesions of allografts at long-term post transplantation. MATERIAL/METHODS: The study group consisted of 274 patients who received a cadaveric kidney allograft between 1994 and 1999. Altogether 553 biopsy specimens were obtained from this group of patients between 1994 and 2004 and graded according to Banff 2005 criteria.Two groups were identified: CS - recipients of kidneys stored in simple hypothermia (n=114) and MP - recipients of kidneys stored by machine perfusion (n=160). There were 161 cadaveric donors, 92 in the Mp group and 69 in the CS group. The 553 biopsy specimens obtained revealed the following: interstitial fibrosis and tubular atrophy - 189, chronic rejection - 19, acute rejection - 64, arteriosclerosis - 117, calcineurin inhibitor toxicity 25, microangiopathy - 44, focal glomerulosclerosis - 82, arteriole hyalinization - 85, ATN - 241. RESULTS: In the CS group histopathological lesions consistent with interstitial fibrosis and tubular atrophy were more frequently encountered than in the MP group (90% vs 64%, p<0.05) Also, chronic rejection was more frequent in the CS group (9% vs 3%, p<0.05). The remaining lesions encountered in biopsies did not differ significantly between the groups. CONCLUSIONS: Kidneys preserved by cold storage are more frequently affected by chronic rejection and interstitial fibrosis. PMID- 19293753 TI - Developing the European Physical and Rehabilitation Medicine Journal's Network. PMID- 19293754 TI - Les Annales de Readaptation et de Medecine Physique publication in 2008. AB - In 2008 the European Society of Physical and Rehabilitation Medicine (ESPRM) established a network of European journal and promoted collaboration among the European Physical and Rehabilitation Medicine (PRM) journals. In this connection, the authors present here the articles published in the Annales de Readaptation et Medecine Physique in 2008. In the same time, the European Journal presents in the Annales de Readaptation et Medecine Physique its main articles of year 2008. The papers published in the Annales de Readaptation et Medecine Physique in 2008 were reviewed, classified by topics and discussed. Sixty-four papers are described, with a coverage of all the fields of PRM, from childhood to elderly, from basic science to clinical research, from case studies to epidemiological works. The aim of this paper was to provide physiatrists with some articles of the Annales de Readaptation et Medecine Physique related to their working condition. Next year, the name of our journal will become Annals of Physical and Rehabilitation Medicine, as most of the articles will be published in French and in English, with the hope that more and more European authors and readers will find it more convenient. PMID- 19293755 TI - International focus on stroke. Foreword. PMID- 19293756 TI - Assessment of sensorized garments as a flexible support to self-administered post stroke physical rehabilitation. AB - The aim of this study was to provide a description of a newly-developed remote rehabilitation system that can be employed both at home and in the hospital, supporting motor rehabilitation for post-stroke patients with upper-limb impairment. A garment, which embeds kinesthetic sensors made of a piezoresitive polymer, is provided with a wireless connection to a computer (the patient station). The station detects in real time whether the patient is performing the exercises correctly or not, and provides feedback through an easy visual representation on the screen. Movement recognition is performed using a template matching approach, which allows exercises to be defined during each session as required without additional configuration. In this study an healthy volunteer used the garment to record 840 exercises, mimicking both correct and incorrect compensatory movements under expert supervision. The sensitivity and specificity of the recognition system were measured through its ability to correctly identify the pre-labelled exercises. A pilot set of 13 post-stroke subjects (mean age of 50) was then offered to use the rehabilitation system while in the neuro rehabilitation ward; the acceptability was assessed through a 10-question subjective evaluation questionnaire. The wearable system tested provided a raw recognition performance (correct-versus-incorrect exercise detection) above 90%. The majority of the patients were satisfied with the system, considered it useful, and would use it at home. In conclusion, computer-based interventions can support widespread, earlier and more intense physical therapy after a neurological event, provided they are easy to use and blend well with the existing rehabilitation workflow. Wearable sensors are promising candidates to realize unobtrusive devices to support the rehabilitation process and its continuity after discharge from the Rehabilitation Unit. PMID- 19293757 TI - EJPRM systematic continuous update on Cochrane reviews in rehabilitation: news from the 4th Issue 2008. AB - AIM: In 2007 a systematic review about the rehabilitation contents of the Cochrane Collaboration was published. That review was then continuously updated by systematically searching the new papers published in every Cochrane Issue. The aim of the present paper was to systematically review all the rehabilitation papers published in the 4th Issue of 2008 from the Cochrane Library. METHODS: The author systematically searched all the paper of rehabilitative interest from the 4th Issue 2008 of the Cochrane Library. The papers have been divided in 3 groups: new reviews, updated reviews and protocols. Each group have been then divided in subgroups on the base of the topic. RESULTS: The number of included papers was 27, including 7 new reviews, 11 updated reviews and 9 protocols. About updated reviews, 4 reviews changed conclusions on the base of the most recent RCTs. CONCLUSIONS: The Cochrane Collaboration and its product, the Cochrane Library are really relevant instruments to improve evidence-based medicine (EBM) in medical practice and thus also in the rehabilitation field. The present paper can help REHABILITATION SPECIALISTS to easily retrieve the conclusions of the most relevant and updated reviews in order to change their clinical practice in a more rapid and effective way. PMID- 19293758 TI - Technology in rehabilitation. PMID- 19293759 TI - The impact of robotic rehabilitation in children with acquired or congenital movement disorders. AB - AIM: The aim of this study was to evaluate if the robot-mediated therapy (RMT) can yield positive outcomes in children with acquired or congenital upper extremity movement disorders. METHODS: This was an uncontrolled pilot study with pre-post treatment outcome comparison carried out by the Pediatric Rehabilitation Department of a Children's Hospital. The study enrolled 12 children, aged 5 to 15 years, suffering from acquired (at least 12 months post-onset) or congenital upper limb motor impairment. ETIOLOGY: 4 stroke, 6 traumatic brain injuries, and 2 hemiplegic cerebral palsy. RMT was provided 3 times a week for an hour during 6 weeks for a total of 18 robot therapy sessions. The Melbourne Scale (MS) and the upper-extremity subsection of the Fugl-Meyer Assessment (FMA) were used for measurement of impairment. Secondary outcome measurements were made through the Modified Ashworth Scale (MAS); the Reaching Performance Scale (RPS); Parent's Questionnaire, and robot-based evaluation measurements. Specifically, authors compared the smoothness, as measured by the jerk metric, and average speed of unconstrained reaching movements. RESULTS: Pre-post clinical evaluation revealed statistically significant gains for all primary and secondary metrics. In addition, significant improvement of robot-based metrics was observed. The primary outcome measurement mean (SEM) gains were 6.71 (1.29) for MS and 3.33 (0.80) for the FMA. RMT led to spasticity decreases in chronic cases, as shown by the reduction of MAS. It led to improved trunk-upper extremity postural attitude as demonstrated by improved RPS, and it was well accepted by parents and children as observed in the Parent's Questionnaire. CONCLUSIONS: This study suggests that RMT may hold rehabilitative benefits in children suffering from acquired and congenital hemiparesis. PMID- 19293761 TI - New year, new PubMed. AB - Aim of this contribution was to present the latest additions to the worldwide known PubMed interface, concerning a better visualization of results list; a new format for the Advanced search which has merged the old labels' options; new features about saving collections and searches via MyNCBI and about creating one's own bibliography. PMID- 19293762 TI - The continuing strategy of the QJNM&MI: a welcome address by the incoming Editor. PMID- 19293763 TI - PET-CT for tailoring therapy of solid tumors. PMID- 19293764 TI - New PET tracers for evaluation of solid tumor response to therapy. AB - We take advantage of the special characteristics of human tumors to image tumor response including predicting susceptibility to specific cancer therapies. The metabolic phenotype of malignancy, includes increased glycolysis (2-[(18)F]fluoro 2-D-deoxyglucose [FDG]), increased proliferation (2-[(18)F]fluoro-L-thymidine [FLT]), increased amino acid transport, as well as other functions such as fatty acid synthesis which have yet to be fully exploited. The endocrine dependent malignancies offer rich opportunities for selective imaging, including radioligands that have high affinity for hormone receptors, like androgen receptor (AR) (16Beta-[(18)F]16beta-[(18)F]fluoro-5alpha-dihydrotestoste-rone [FDHT]) and estrogen receptor (ER) ([(18)F]fluoroestradiol [FES]) and tissue specific transporters such as sodium iodide symporter (NIS) ((124)I). As knowledge of cancer biology has grown, it has become possible to develop tracers which image the client proteins involved in response to specific drugs, e.g. Gallium-68-Fab'2 herceptin for HER 2 effected by HSP 90 inhibitor drugs. More and more radiolabeled drugs will be used to explore the pharmacology of anticancer therapies, such as [(18)F]Desatinib. These may or may not be excellent imaging agents, but as analogs or even true tracers for specific anti-cancer drugs, noninvasive imaging through positron emission tomography will provide highly useful information, relating cancer pharmacology within the local tumor, to treatment response. PMID- 19293765 TI - [18F]FDG-PET-CT for early monitoring of tumor response: when and why. AB - (18)F-fluorodeoxyglucose positron emission tomography ([(18)F]FDG PET) has been recognized as a suitable tool in tumor response assessment of patients complaining with solid tumors who have undergone chemo- and radiotherapy. It offers the advantage of functional tissue characterization, which is independent from morphologic criteria allowing to differentiate disease relapse from therapy induced fibrosis. At present, there is a growing body of evidence that PET semi quantitative assessment of treatment-induced changes in tumor [(18)F]FDG avidity may predict early tumor response and patient outcome. Patient management might be changed. For instance, in non responder patients this novel diagnostic approach would hamper useless ''wait and watch'' attitude in implementing further options or identifying those needing additional therapeutic strategies. On the other hand, for those patients revealing promptly a favourable metabolic response a cost-sparing approach could be implemented avoiding expensive diagnostic procedures during the follow-up as well as the risk of over-treating. In any case, since even a partial metabolic response may be an indication for continuing therapy, the advantage of metabolic assessment over conventional procedures may be clinically relevant. Although a morphological assessment has been considered for long time the standard for detecting therapy response, limitations of conventional computed tomography-based evaluation in solid tumors are well-known. PET provides an independent means of assessing malignancy. However, no consensus has been achieved until now regarding the optimal timing in performing PET during or at completion of treatment. PMID- 19293766 TI - Molecular imaging to select cancer therapy and evaluate treatment response. AB - The ability to assay in vivo biologic processes non-invasively and quantitatively makes molecular imaging a particularly attractive tool for clinical trials of new drugs and for clinical cancer practice. This review highlights the emerging application of molecular imaging to cancer drug testing and clinical cancer treatment. The potential roles that imaging can play in the approach to cancer drug trials and clinical treatment are first highlighted, including applications to early drug testing, Phase II and III clinical trials, and clinical practice. The use of molecular imaging to select cancer therapy is then discussed, citing examples where molecular imaging can be used to measure the expression of therapeutic targets and factors mediating therapeutic resistance. The use of imaging to measure early pharmacodynamic changes and subsequent response to cancer treatment is then reviewed. Finally, the need for standardization and reproducible quantitative imaging analysis is reviewed. PMID- 19293767 TI - Assessing tumor hypoxia by positron emission tomography with Cu-ATSM. AB - For the last several decades, hypoxia has been recognized to be one of the key factors in tumor aggression and an important impediment to local and distant control of malignant tumors. In addition, hypoxia is a major cause of failure of both radiation therapy and chemotherapy. It has been shown that hypoxia is an independent negative prognostic factor for patient outcome in various solid tumors. Clinical studies using polarographic oxygen electrodes, as a tool for measuring hypoxia, were the first to demonstrate the presence of hypoxia in human tumors and its association with poor prognosis. However, this method is invasive and has technical limitations that prevent its routine clinical use. Over the years, imaging as a noninvasive method has attracted a lot of attention and several radiotracers have been developed for noninvasive evaluation of hypoxia. One of the most promising radiotracers is the copper(II) complex of diacetyl-2,3 bis(N(4)-methyl-3-thiosemicarbazonato) ligand (Cu-ATSM) for imaging with positron emission tomography. In this review, the preclinical evaluation of Cu-ATSM as well as its clinical value in several solid tumors will be discussed. PMID- 19293768 TI - The evolving role of PET-CT in the management of esophageal cancer. AB - Esophageal cancer, although uncommon in the United States, has been increasing in frequency, and despite advances in therapy, the diagnosis still carries a poor prognosis. Many patients present with locally advanced disease and receive multimodality therapy with combined chemoradiation before surgery. Imaging plays an important role in the initial selection of patients for tri-modality therapy and in evaluating responses to neoadjuvant chemoradiation prior to surgery. There is increasing use of 2-deoxy-2-[(18)F]fluoro-D-glucose (FDG)-positron emission tomography (PET)/computed tomography (CT) in management of esophageal cancer to identify distant metastasis at presentation and to assess response to therapy. Standardized imaging protocols with semiquantitative analysis may also provide prognostic information and be used to individualize patient therapy. This review will discuss the role of imaging studies in the management of esophageal cancer patients, with particular attention to FDG-PET/CT. PMID- 19293769 TI - Breast cancer therapy: the role of PET-CT in decision making. AB - This paper describes the role of positron emission tomography (PET) and PET computed tomography (CT) in breast cancer patients. Fluorine-18-Fluoro-D-glucose (FDG) has limited diagnostic value in detecting small noninvasive primary tumors, in staging the axillary region in early stages and in the detection of osteoblastic metastases. Better results have been shown in the detection and staging of primary invasive tumors. Significant clinical data are available in the monitoring of primary chemotherapy in locally advanced breast cancer where [(18)F]FDG PET-CT allows prediction of the response even shortly after the onset of therapy. Quantitative evaluation of tumor uptake is necessary. Therapy-induced changes in tumor metabolism may be helpful in making decisions about continuation, modification or cessation of therapy. Therefore, [(18)F]FDG PET-CT appears to be a promising tool for the personalization of breast cancer treatment by its early identification of nonresponders. It offers improved patient care, avoiding ineffective chemotherapy and the side effects while reducing the cost. An area generating high expectations for PET-TC in breast cancer is in monitoring in order to tailor therapy to the tumor characteristics of individual patients who may require tracers other than [(18)F]FDG. The introduction of new PET tracers and the development of new instruments will offer opportunities to improve the role of PET-CT in decision making of therapy in these patients. PMID- 19293770 TI - Tailoring therapy in colorectal cancer by PET-CT. AB - Positron emission tomography (PET) using [(18)F]-fluoro-2'-deoxy-D-glucose (FDG) has an added value in the clinical management of patients with colorectal carcinoma (CRC). This includes restaging patients before surgical resection or local recurrence of liver metastases, assessment whether residual lesions are scar or recurrence and in pinpointing recurrence in case of unexplained increase in serum levels of carcinoembryonic antigen. At present, there is an increasing interest in new roles for FDG-PET, especially for characterization of lesions, for prognosis and response prediction and for early evaluation of treatment response to commenced therapy. FDG-PET may lead to better selection of patients for different therapeutic options or to early individual adjustment of current treatment. This systematic review aims to provide an up-to-date overview of literature on the current and potential value of FDG-PET in CRC patients by addressing staging and recurrence detection, prognosis and response prediction and evaluation of preoperative (chemo)radiotherapy for primary rectal carcinoma, ablative treatment for unresectable liver metastases and chemotherapy for advanced CRC. PMID- 19293771 TI - PET-CT for treatment planning in prostate cancer. AB - Molecular imaging techniques, such as positron emission tomography (PET), may be of help in management treatment planning. In particular, in prostate cancer patients, PET and PET-computed tomography (PET-CT) can be successfully used in treatment planning at different steps, including: 1) tumor characterization and staging, to define the most appropriate primary treatment; 2) re-staging, to define a second line therapy on the site of possible recurrences; and 3) monitoring the disease and the efficacy of treatment. Although the most commonly used PET tracer, [(18)F]Fluorodeoxyglucose ([(18)F]FDG), presents limitations in imaging prostate cancer patients, several alternative PET tracers have been proposed to evaluate by PET these patients, with promising RESULTS: Optimal treatment for prostate cancer depends on the accuracy in tumor characterization and staging. In fact, localized primary tumor can be treated with radical prostatectomy, while metastatic tumor is usually treated with systemic therapeutic regimen. Different PET tracers, including [(11)C]Choline, [(18)F]Choline and [(11)C]Acetate, have been successfully reported. Howe-ver, further studies in large population of patients are still necessary to establish their final clinical role in the primary detection and staging of prostate cancer. The information on the site of possible recurrences is also important for therapeutic strategies. Several PET tracers have been proposed to re-stage prostate cancer patients. In particular, [11C]Choline PET has now been established as a clinical procedure to non-invasively re-stage, in a single session, prostate cancer patients presenting an increase of prostate specific antigen (PSA) after radical treatment. The role of PET and PET-CT in monitoring the disease and the effects of treatment are under investigation and still to be defined. In the present review, we focused on the use of several PET tracers in different clinical indications aimed at the treatment planning of prostate cancer patients. PMID- 19293772 TI - Oncolytic reovirus effectively targets breast cancer stem cells. AB - Recent evidence suggests that cancer stem cells (CSCs) play an important role in cancer, as these cells possess enhanced tumor-forming capabilities and are resistant to current anticancer therapies. Hence, novel cancer therapies will need to be tested for both tumor regression and CSC targeting. Herein we show that oncolytic reovirus that induces regression of human breast cancer primary tumor samples xenografted in immunocompromised mice also effectively targets and kills CSCs in these tumors. CSCs were identified based on CD24(-)CD44(+) cell surface expression and overexpression of aldehyde dehydrogenase. Upon reovirus treatment, the CSC population was reduced at the same rate as non-CSCs within the tumor. Immunofluorescence of breast tumor tissue samples from the reovirus- and mock-treated mice confirmed that both CSCs and non-CSCs were infectible by reovirus, and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) assay showed that both populations died by apoptosis. Ras, which has been shown to mediate reovirus oncolysis, was found to be present at similar levels in all cell types, and this is consistent with their comparable sensitivity to reovirus. These experiments indicate that oncolytic reovirus has the potential to induce tumor regression in breast cancer patients. More important, the CSC population was equally reduced and was as susceptible to reovirus treatment as the non-CSC population. PMID- 19293773 TI - Host and vector-dependent effects on the risk of germline transmission of AAV vectors. AB - The assessment of the risk of germline transmission of vector-coded sequences is critical for clinical translation of gene transfer strategies. We used rabbit models to analyze the risk of germline transmission of adeno-associated viral (AAV) vectors. Intravenous injection of AAV-2 or AAV-8 resulted in liver mediated, long-term expression of therapeutic levels of human factor IX (hFIX) in a dose-dependent manner. In high-dose cohorts, AAV-8 resulted in twofold higher levels of circulating hFIX and of vector DNA in liver compared to AAV-2. Vector sequences were found in the semen of all rabbits. The kinetics of vector clearance from semen was dose- and time-dependent but serotype-independent. No late recurrence of AAV-8 sequences was found in the semen over several consecutive cycles of spermatogenesis. In a novel rabbit model, AAV-2 or AAV-8 sequences were detected in the semen of vasectomized animals that lack germ cells. Therefore, structures of the genitourinary (GU) tract, as well as the testis, contribute significantly to vector shedding in the semen. Collectively, data from these two models suggest that the risk of inadvertent germline transmission in males by AAV-8 vectors is low, similar to that of AAV-2, and that AAV dissemination to the semen is in part modulated by host-dependent factors. PMID- 19293774 TI - The pharmacological chaperone N-butyldeoxynojirimycin enhances enzyme replacement therapy in Pompe disease fibroblasts. AB - In spite of the progress in the treatment of lysosomal storage diseases (LSDs), in some of these disorders the available therapies show limited efficacy and a need exists to identify novel therapeutic strategies. We studied the combination of enzyme replacement and enzyme enhancement by pharmacological chaperones in Pompe disease (PD), a metabolic myopathy caused by the deficiency of the lysosomal acid alpha-glucosidase. We showed that coincubation of Pompe fibroblasts with recombinant human alpha-glucosidase and the chaperone N butyldeoxynojirimycin (NB-DNJ) resulted in more efficient correction of enzyme activity. The chaperone improved alpha-glucosidase delivery to lysosomes, enhanced enzyme maturation, and increased enzyme stability. Improved enzyme correction was also found in vivo in a mouse model of PD treated with coadministration of single infusions of recombinant human alpha-glucosidase and oral NB-DNJ. The enhancing effect of chaperones on recombinant enzymes was also observed in fibroblasts from another lysosomal disease, Fabry disease, treated with recombinant alpha-galactosidase A and the specific chaperone 1 deoxygalactonojirimycin (DGJ). These results have important clinical implications, as they demonstrate synergy between pharmacological chaperones and enzyme replacement. A synergistic effect of these treatments may result particularly useful in patients responding poorly to therapy and in tissues in which sufficient enzyme levels are difficult to obtain. PMID- 19293775 TI - HSV-mediated transfer of artemin overcomes myelin inhibition to improve outcome after spinal cord injury. AB - Artemin is a neurotrophic factor of the glial cell line-derived neurotrophic factor (GDNF) family of ligands that acts through the GDNF family receptor alpha3 (GFRalpha3)/ret receptor found predominantly on sensory and sympathetic neurons. In order to explore the potential utility of artemin to improve functional outcome after spinal cord injury (SCI), we constructed a nonreplicating herpes simplex virus (HSV)-based vector to express artemin (QHArt). We found that QHArt efficiently transfects spinal cord neurons to produce artemin. Transgene-mediated artemin supported the extension of neurites by primary dorsal root ganglion neurons in culture, and allowed those cells to overcome myelin inhibition of neurite extension through activation of protein kinase A (PKA) to phosphorylate cyclic adenosine monophosphate (cAMP) response element binding protein (CREB) and increase expression of arginase I. Intraspinal injection of QHArt immediately after thoracic spinal cord dorsal over hemisection produced a statistically significant improvement in motor recovery over the course of four weeks measured by locomotor rating score. PMID- 19293776 TI - Rational design of antisense oligomers to induce dystrophin exon skipping. AB - Duchenne muscular dystrophy (DMD), one of the most severe neuromuscular disorders of childhood, is caused by the absence of a functional dystrophin. Antisense oligomer (AO) induced exon skipping is being investigated to restore functional dystrophin expression in models of muscular dystrophy and DMD patients. One of the major challenges will be in the development of clinically relevant oligomers and exon skipping strategies to address many different mutations. Various models, including cell-free extracts, cells transfected with artificial constructs, or mice with a human transgene, have been proposed as tools to facilitate oligomer design. Despite strong sequence homology between the human and mouse dystrophin genes, directing an oligomer to the same motifs in both species does not always induce comparable exon skipping. We report substantially different levels of exon skipping induced in normal and dystrophic human myogenic cell lines and propose that animal models or artificial assay systems useful in initial studies may be of limited relevance in designing the most efficient compounds to induce targeted skipping of human dystrophin exons for therapeutic outcomes. PMID- 19293777 TI - Regulated and multiple miRNA and siRNA delivery into primary cells by a lentiviral platform. AB - RNA interference (RNAi) has tremendous potential for investigating gene function and developing new therapies. However, the design and validation of proficient vehicles for stable and safe microRNA (miR) and small interfering RNA (siRNA) delivery into relevant target cells remains an active area of investigation. Here, we developed a lentiviral platform to efficiently coexpress one or more natural/artificial miR together with a gene of interest from constitutive or regulated polymerase-II (Pol-II) promoters. By swapping the stem-loop (sl) sequence of a selected primary transcript (pri-miR) with that of other miR or replacing the stem with an siRNA of choice, we consistently obtained robust expression of the chimeric/artificial miR in several cell types. We validated our platform transducing a panel of engineered cells stably expressing sensitive reporters for miR activity and on a natural target. This approach allowed us to quantitatively assess at steady state the target suppression activity and expression level of each delivered miR and to compare it to those of endogenous miR. Exogenous/artificial miR reached the concentration and activity typical of highly expressed natural miR without perturbing endogenous miR maturation or regulation. Finally, we demonstrate the robust performance of the platform reversing the anergic/suppressive phenotype of human primary regulatory T cells (Treg) by knocking-down their master gene Forkhead Transcription Factor P3 (FOXP3). PMID- 19293778 TI - Transcriptional enhancers induce insertional gene deregulation independently from the vector type and design. AB - The integration characteristics of retroviral (RV) vectors increase the probability of interfering with the regulation of cellular genes, and account for a tangible risk of insertional mutagenesis in treated patients. To assess the potential genotoxic risk of conventional or self-inactivating (SIN) gamma-RV and lentiviral (LV) vectors independently from the biological consequences of the insertion event, we developed a quantitative assay based on real-time reverse transcriptase--PCR on low-density arrays to evaluate alterations of gene expression in individual primary T-cell clones. We show that the Moloney leukemia virus long terminal repeat (LTR) enhancer has the strongest activity in both a gamma-RV and a LV vector context, while an internal cellular promoter induces deregulation of gene expression less frequently, at a shorter range and to a lower extent in both vector types. Downregulation of gene expression was observed only in the context of LV vectors. This study indicates that insertional gene activation is determined by the characteristics of the transcriptional regulatory elements carried by the vector, and is largely independent from the vector type or design. PMID- 19293779 TI - Increased expression of catalase and superoxide dismutase 2 reduces cone cell death in retinitis pigmentosa. AB - Oxidative and nitrosative damage are major contributors to cone cell death in retinitis pigmentosa (RP). In this study, we explored the effects of augmenting components of the endogenous antioxidant defense system in models of RP, rd1, and rd10 mice. Unexpectedly, overexpression of superoxide dismutase 1 (SOD1) in rd1 mice increased oxidative damage and accelerated cone cell death. With an elaborate mating scheme, genetically engineered rd10 mice with either inducible expression of SOD2, Catalase, or both in photoreceptor mitochondria were generated. Littermates with the same genetic background that did not have increased expression of SOD2 nor Catalase provided ideal controls. Coexpression of SOD2 and Catalase, but not either alone, significantly reduced oxidative damage in the retinas of postnatal day (P) 50 rd10 mice as measured by protein carbonyl content. Cone density was significantly greater in P50 rd10 mice with coexpression of SOD2 and Catalase together than rd10 mice that expressed SOD2 or Catalase alone, or expressed neither. Coexpression of SOD2 and Catalase in rd10 mice did not slow rod cell death. These data support the concept of bolstering the endogenous antioxidant defense system as a gene-based treatment strategy for RP, and also indicate that coexpression of multiple components may be needed. PMID- 19293780 TI - Metaplastic esophageal columnar epithelium without goblet cells shows DNA content abnormalities similar to goblet cell-containing epithelium. AB - OBJECTIVES: The mucosa of patients with columnar-lined esophagus recognized on endoscopy usually shows epithelium with and without goblet cells. Columnar epithelium with goblet cells ("Barrett's esophagus") is generally believed to represent a premalignant lesion and has been shown to contain DNA abnormalities. However, the biological properties of non-goblet columnar epithelium remain unknown. The purpose of this study was to determine the DNA content properties of non-goblet epithelium in patients with metaplastic columnar epithelium of the esophagus. METHODS: Mucosal biopsies of the esophagus from 68 patients with columnar metaplasia of the esophagus (22 without goblet cells and 46 with goblet cells) and 19 patients with normal gastric mucosa (controls) were histologically evaluated for the density of goblet cells. The latter group was divided into low density, high-density, and very high-density goblet cell subgroups. Tissue sections of non-goblet epithelium and goblet cell epithelium (where present) were evaluated by image cytometry, and high-fidelity DNA histograms were created to indicate the G0/G1 peak DNA index (DI), DNA content heterogeneity index (HI), and the percentage of cells with DNA exceeding 5N (5N-EC). G0/G1 peaks with DI>1.1 were considered aneuploid. RESULTS: Normal gastric controls showed a mean peak DI of 1.02+/-0.03 and an HI of 11.6+/-0.7. None of the controls revealed aneuploidy or 5N-EC. Patients with metaplastic columnar epithelium with goblet cells showed a DI of 1.15+/-0.12, HI of 18.2+/-2.1, mild aneuploidy in 54% of the cases, and 5N-EC in 15% of the cases, all of which were significantly higher than in controls. Patients with metaplastic columnar epithelium without goblet cells showed DNA content results statistically similar to those of patients with metaplastic columnar epithelium with goblet cells, and also revealed significantly higher values compared with those of controls. Furthermore, there were no significant differences in any of the key DNA content abnormalities between non-goblet and goblet cell-containing epithelium in patients with metaplastic columnar epithelium with goblet cells, or between these two types of epithelium according to the density of goblet cells. CONCLUSIONS: DNA content abnormalities occur with equal frequency and extent in metaplastic columnar epithelium of the esophagus without goblet cells compared with metaplastic columnar epithelium with goblet cells. These findings suggest that metaplastic non-goblet columnar epithelium of the esophagus may have neoplastic potential. PMID- 19293781 TI - Natural history of minimal hepatic encephalopathy in patients with extrahepatic portal vein obstruction. AB - OBJECTIVES: Minimal hepatic encephalopathy (MHE) leads to deterioration in patient quality of life and could be a marker for future episodes of clinical hepatic encephalopathy (HE) in liver cirrhosis. Whether MHE predicts HE in extrahepatic portal vein obstruction (EHPVO) is not known. We studied the incidence of overt HE in EHPVO patients with MHE. METHODS: Consecutive patients (from October 2006 to July 2007) with a diagnosis of EHPVO were followed up at 3 month intervals. MHE was diagnosed by abnormal psychometry (>2 s.d.) and/or P300 auditory event-related potential (P300 ERP) (>2.5 s.d.), and HE was diagnosed as per West-Heaven criteria. Critical flicker frequency (CFF) was also measured at baseline and after 1 year. RESULTS: Thirty-two EHPVO patients (age, 23.2+/-10.8 years; M/F 22:10) were followed up for 1 year. Of 32 patients, P300 ERP was prolonged in 8 (25%) (371.8+/-13.9 ms), 9 (28%) had abnormal psychometric tests, and CFF was <38 Hz in 8 (25%) patients after a follow-up of 13.5+/-2.4 months. Of 12 patients who had MHE at baseline, 9 (75%) patients continued to have MHE, and in 3 (25%) patients it disappeared. One (5%) of the remaining 20 patients developed MHE during the follow-up. Venous ammonia level was higher in patients with MHE (79.7+/-17.0 micromol/l; range 33-124) compared with patients without MHE (46.6+/-19.8 micromol/l; range 24-78, P<0.001) on follow-up. Similarly, patients who had spontaneous shunts (n=10) had significantly higher venous ammonia levels (82.4+/-20.3 vs. 47.1+/-16.7 micromol/l; P=0.001) than those who had no shunt (n=22). Neither patients who had MHE nor those who did not have MHE at baseline developed HE. CONCLUSIONS: Seventy-five percent of extrahepatic portal vein obstruction patients with MHE continued to have MHE, and new-onset MHE developed in 5% over 1 year. In this small sample, patients with EHPVO and MHE did not progress to overt encephalopathy within the relatively short time frame studied. PMID- 19293782 TI - NAFLD as a risk factor for the development of diabetes and the metabolic syndrome: an eleven-year follow-up study. AB - OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) uncommonly results in cirrhosis and liver-related death; however, its impact on the development of metabolic complications remains unclear. We sought to determine whether NAFLD with elevated aminotransaminase (ALT) levels was a risk factor for incident diabetes or the metabolic syndrome (MS) over an 11-year period. METHODS: Adult residents of Busselton, Western Australia underwent assessment in 1994-1995 as part of the Busselton Health Survey. NAFLD was diagnosed on the basis of a raised ALT (>40 IU/l) after the exclusion of alcohol, viral, metabolic, and autoimmune liver disease. NAFLD and non-NAFLD subjects were reassessed in 2005 for liver complications, diabetes, and the MS. RESULTS: A total of 358 subjects, 68% male (109 NAFLD, 249 non-NAFLD), mean age (s.d.) 59.9 (11.6) years, attended follow-up 11.1 years after the initial assessment. After excluding subjects with diabetes at baseline, those with NAFLD were more likely to have developed diabetes on follow-up (20/106, 18.9% vs. 15/246, 6.1%; P<0.001). After excluding subjects with MS at baseline, those with NAFLD were more likely to have developed MS at follow-up (27/81, 33.3% vs. 51/226, 22.6%; P=0.056). However, in multivariate logistic regression models, NAFLD was no longer a significant independent predictor of the development of diabetes or MS after adjusting for baseline waist circumference, hypertension, and insulin resistance. None of the subjects developed liver complications. CONCLUSIONS: Subjects with NAFLD and elevated ALT levels are at an increased risk of developing diabetes and the MS. This may be because of the presence of associated metabolic risk factors. PMID- 19293783 TI - Thalidomide as a treatment for refractory CGD colitis. PMID- 19293784 TI - Adequate relief in a treatment trial with IBS patients: a prospective assessment. AB - OBJECTIVES: Adequate relief (AR) of irritable bowel syndrome (IBS) symptoms (IBS AR) has been used as a primary end point in many randomized controlled trials of IBS and is considered by the Rome III committee to be an acceptable primary end point. However, controversy exists on whether baseline severity confounds the effect of the treatment outcome. The aim (1) is to compare a subjective report of IBS-AR with global assessment of improvement (IBS-GAI), change in IBS symptom severity scale (IBS-SSS), and IBS quality of life (IBS-QOL); (2) to explore whether initial IBS symptom severity influences the ability of these outcome measures to detect differences post treatment; and (3) to determine whether psychological symptoms influence the sensitivity of these measures, in a randomized controlled treatment trial. METHODS: A total of 289 adult IBS patients were recruited to a treatment trial. Baseline IBS-SSS scores were used to classify IBS severity as mild (<175), moderate (175-300), or severe (>300). Questionnaires were completed at baseline and after 3 weeks of treatment with sham acupuncture or wait-list control. RESULTS: IBS baseline severity (IBS-SSS) significantly affected the proportion of patients who reported IBS-AR at 3 weeks (mild, 70%; moderate, 49.7%; severe, 38.8%) (P<0.05). However, once the patients who reported IBS-AR at baseline (28.0%) were excluded from the analysis, baseline severity no longer affected the proportion of patients reporting IBS-AR. Baseline severity did not have a significant effect on patients reporting moderate or significant improvement on the IBS-GAI (mild, 30%; moderate, 25.3%; severe, 18.8%) (P=NS). Psychological symptoms had no significant correlations with responders after adjusting for baseline severity. CONCLUSIONS: These data suggest that IBS-AR as an end point is inversely related to baseline symptom severity. However, if patients who report AR at screening were excluded from study participation, baseline symptom severity was no longer confounded with a report of AR at the study end point. PMID- 19293785 TI - A rare cause of large liver cysts. PMID- 19293786 TI - Using a multifaceted approach to improve the follow-up of positive fecal occult blood test results. AB - OBJECTIVES: Inadequate follow-up of abnormal fecal occult blood test (FOBT) results occurs in several types of practice settings. Our institution implemented multifaceted quality improvement (QI) activities in 2004-2005 to improve follow up of FOBT-positive results. Activities addressed precolonoscopy referral processes and system-level factors such as electronic communication, provider education, and feedback. We evaluated their effects on timeliness and appropriateness of positive-FOBT follow-up and identified factors that affect colonoscopy performance. METHODS: Retrospective electronic medical record review was used to determine outcomes before and after QI activities in a multispecialty ambulatory clinic of a tertiary care Veterans Affairs facility and its affiliated satellite clinics. From 1869 FOBT-positive cases, 800 were randomly selected from time periods before and after QI activities. Two reviewers used a pretested standardized data collection form to determine whether colonoscopy was appropriate or indicated based on predetermined criteria and if so, the timeliness of colonoscopy referral and performance before and after QI activities. RESULTS: In cases where a colonoscopy was indicated, the proportion of patients who received a timely colonoscopy referral and performance were significantly higher post-implementation (60.5% vs. 31.7%, P<0.0001 and 11.4% vs. 3.4%, P=0.0005). A significant decrease also resulted in median times to referral and performance (6 vs. 19 days, P<0.0001 and 96.5 vs. 190 days, P<0.0001) and in the proportion of positive-FOBT test results that had received no follow-up by the time of chart review (24.3% vs. 35.9%, P=0.0045). Significant predictors of absence of the performance of an indicated colonoscopy included performance of a non-colonoscopy procedure such as barium enema or flexible sigmoidoscopy (OR=16.9; 95% CI, 1.9-145.1), patient non-adherence (OR=33.9; 95% CI, 17.3-66.6), not providing an appropriate provisional diagnosis on the consultation (OR=17.9; 95% CI, 11.3-28.1), and gastroenterology service not rescheduling colonoscopies after an initial cancellation (OR=11.0; 95% CI, 5.1-23.7). CONCLUSIONS: Multifaceted QI activities improved rates of timely colonoscopy referral and performance in an electronic medical record system. However, colonoscopy was not indicated in over one third of patients with positive FOBTs, raising concerns about current screening practices and the appropriate denominator used for performance measurement standards related to colon cancer screening. PMID- 19293787 TI - A prospective evaluation of the bedside index for severity in acute pancreatitis score in assessing mortality and intermediate markers of severity in acute pancreatitis. AB - OBJECTIVES: Our aim was to prospectively evaluate the ability of the bedside index for severity in acute pancreatitis (BISAP) score to predict mortality as well as intermediate markers of severity in a tertiary center. METHODS: The BISAP score was evaluated among 397 consecutive cases of acute pancreatitis admitted to our institution between June 2005 and December 2007. BISAP scores were calculated on all cases using data within 24 h of presentation. The ability of the BISAP score to predict mortality was evaluated using trend and discrimination analysis. The optimal cutoff score for mortality from the receiver operating curve was used to evaluate the development of organ failure, persistent organ failure, and pancreatic necrosis. RESULTS: Among 397 cases, there were 14 (3.5%) deaths. There was a statistically significant trend for increasing mortality (P < 0.0001) with increasing BISAP score. The area under the receiver operating curve for mortality by BISAP score in the prospective cohort was 0.82 (95% confidence interval: 0.70, 0.95), which was similar to that of the previously published validation cohort. A BISAP score >or=3 was associated with an increased risk of developing organ failure (odds ratio=7.4, 95% confidence interval: 2.8, 19.5), persistent organ failure (odds ratio=12.7, 95% confidence interval: 4.7, 33.9), and pancreatic necrosis (odds ratio=3.8, 95% confidence interval: 1.8, 8.5). CONCLUSIONS: The BISAP score represents a simple way to identify patients at risk of increased mortality and the development of intermediate markers of severity within 24 h of presentation. This risk stratification capability can be utilized to improve clinical care and facilitate enrollment in clinical trials. PMID- 19293788 TI - Hypertriglyceridemic pancreatitis: presentation and management. AB - Hypertriglyceridemia (HTG) is reported to cause 1-4% of acute pancreatitis (AP) episodes. HTG is also implicated in more than half of gestational pancreatitis cases. Disorders of lipoprotein metabolism are conventionally divided into primary (genetic) and secondary causes, including diabetes, hypothyroidism, and obesity. Serum triglyceride (TG) levels above 1,000 mg/dl are usually considered necessary to ascribe causation for AP. The mechanism for hypertriglyceridemic pancreatitis (HTGP) is postulated to involve hydrolysis of TG by pancreatic lipase and release of free fatty acids that induce free radical damage. Multiple small studies on HTGP management have evaluated the use of insulin, heparin, or both. Many series have also reported use of apheresis to reduce TG levels. Subsequent control of HTG with dietary restrictions, antihyperlipidemic agents, and even regular apheresis has been shown anecdotally in case series to prevent future episodes of AP. However, large multicenter studies are needed to optimize future management guidelines for patients with HTGP. PMID- 19293789 TI - Editorial: is adequate relief fatally flawed or adequate as an end point in irritable bowel syndrome? AB - There is controversy on the validity of binary end points used in irritable bowel syndrome (IBS) clinical trials. In a usual-care observational study, baseline severity influenced the response measured as satisfactory relief. This editorial reviews the observations from a non-pharmacological study to assess the effect of baseline severity on the performance of binary end points in large drug trials. The pivotal finding is that once the patients who reported adequate relief at baseline were excluded from the analysis, baseline severity no longer affected the proportion of patients reporting adequate relief of IBS with treatment. As large drug trials enriched the study cohorts for at least moderate severity after a no-treatment, run-in period, it seems likely that the precaution of excluding mild disease de facto resolved the hypothetical weakness of the adequate relief end point. Given the high responsiveness and longitudinal construct validity demonstrated with adequate relief end point, it should be accepted as a trial end point. PMID- 19293790 TI - Editorial: It's time to make organized colorectal cancer screening convenient and easy for patients. AB - Colorectal cancer (CRC) screening is widely recommended, but underused. To increase screening rates, we need to implement organized and population-based systems to promote CRC screening among people in a single region, health plan, or health system. This is ideally accomplished using fecal immunochemical tests (FITs), which can be sent through a mass mailing. The study by Levi and colleagues shows that patients using aspirin, non-steroidal anti-inflammatory drugs (NSAIDs), anti-platelet agents, or anti-coagulants do not need to stop these medications while doing the fecal collection. This makes the testing more convenient for patients, and avoids the risk of adverse cardiovascular events caused by stopping these medications. PMID- 19293791 TI - Evaluation of liver biopsy in Egyptian HBeAg-negative chronic hepatitis B patients at initial presentation: implications for therapy. AB - OBJECTIVES: A subgroup of HBeAg-negative chronic hepatitis B (CHB) patients with alanine aminotransferase (ALT) and/or hepatitis B virus (HBV)-DNA levels below the cutoff values of international guidelines may have significant liver disease and miss the opportunity for early treatment. Histopathological changes of HBeAg negative CHB patients at initial presentation irrespective of HBV-DNA and/or ALT levels to increase the likelihood of patients for treatment are evaluated. METHODS: CHB patients attending Cairo Liver Center from January 2006 to May 2008 had biochemical, serological, and virological screening as well as liver biopsy that was assessed by Metavir score. RESULTS: Fifty-two HBeAg-negative CHB patients (46 male and 6 female) with a median age of 37.5 years were included in the study. Significant fibrosis (>or=F2) was found in 26% (5/19) of patients with serum HBV-DNA <2,000 IU/ml, and 53% (21/40) of patients with ALT level <2xULN. Liver biopsy increased candidacy for treatment by nearly 25% before implementation of the recommended lower ALT levels (30 U/l for male and 19 U/l for female patients), and by 21.2% after implementation of the lower ALT level. Implementation of the lower ALT level increased the candidacy of patients for treatment by 4% (two patients), whereas liver biopsy increased eligibility for treatment by 55.8 % (27/49). CONCLUSIONS: Liver biopsy is more reliable than either ALT or HBV-DNA levels in the decision to treat Egyptian HBeAg-negative CHB patients, even with the implementation of the recommended lower ALT levels. PMID- 19293792 TI - Sensitivity, but not specificity, of a quantitative immunochemical fecal occult blood test for neoplasia is slightly increased by the use of low-dose aspirin, NSAIDs, and anticoagulants. AB - OBJECTIVES: We evaluated the effect of the use of aspirin, nonsteroidal anti inflammatory drugs (NSAIDS), and anticoagulants on the performance of immunochemical fecal occult blood test (I-FOBT). METHODS: A prospective, cross sectional study of 1,221 ambulatory patients having total colonoscopy after preparing three I-FOBTs. Information regarding the use of medications was collected from the health medical organization (HMO) database. I-FOBT was analyzed with the OC-MICRO instrument using both >or=75 and 100 ngHb/ml of buffer thresholds to determine positivity. RESULTS: Colorectal cancer (CRC) was found in 17 and advanced adenomatous polyp (AAP) in 97 patients. A total of 212 patients were using aspirin/NSAIDS at the time of I-FOBT testing. Qualitative analysis for the detection of AAP/CRC reveals a trend for an increased sensitivity with aspirin/NSAIDS use. At the threshold 75 ng/ml for positivity, the sensitivity for the detection of AAP/CRC was 66.7% for aspirin/NSAIDS use vs. 51.2% for nondrug takers (P=0.20), and at the threshold of 100 ng/ml, the sensitivity was 66.7 vs. 46.5% (P=0.09). The specificity, however, was not affected by the use of aspirin/NSAIDS. At the threshold of 75 ng/ml for positivity, the specificity for the detection of AAP/CRC was 89.5% for aspirin/NSAIDS use vs. 91.2% for nondrug takers (P=0.47), and at the threshold of 100 ng/ml, the specificity was 92.17 vs. 93.0% (P=0.69). A total of 33 patients were using antithrombotics/coagulants at the time of I-FOBT testing. This group was small; however, it appears that their use was also associated with a trend for increased sensitivity and no change in specificity. CONCLUSIONS: The use of aspirin/NSAIDS and anticoagulants was associated with a trend for increased sensitivity with no change in specificity for the detection of AAP/CRC. This study suggests that there is no need to stop these agents before I-FOBT testing. PMID- 19293793 TI - An investigation of WNT pathway activation and association with survival in central nervous system primitive neuroectodermal tumours (CNS PNET). AB - Central nervous system primitive neuroectodermal tumours (CNS PNET) are high grade, predominantly paediatric, brain tumours. Previously they have been grouped with medulloblastomas owing to their histological similarities. The WNT/beta catenin pathway has been implicated in many tumour types, including medulloblastoma. On pathway activation beta-catenin (CTNNB1) translocates to the nucleus, where it induces transcription of target genes. It is commonly upregulated in tumours by mutations in the key pathway components APC and CTNNB1. WNT/beta-catenin pathway status was investigated by immunohistochemical analysis of CTNNB1 and the pathway target cyclin D1 (CCND1) in 49 CNS PNETs and 46 medulloblastomas. The mutational status of APC and CTNNB1 (beta-catenin) was investigated in 33 CNS PNETs and 22 medulloblastomas. CTNNB1 nuclear localisation was seen in 36% of CNS PNETs and 27% of medulloblastomas. A significant correlation was found between CTNNB1 nuclear localisation and CCND1 levels. Mutations in CTNNB1 were identified in 4% of CNS PNETs and 20% of medulloblastomas. No mutations were identified in APC. A potential link between the level of nuclear staining and a better prognosis was identified in the CNS PNETs, suggesting that the extent of pathway activation is linked to outcome. The results suggest that the WNT/beta-catenin pathway plays an important role in the pathogenesis of CNS PNETs. However, activation is not caused by mutations in CTNNB1 or APC in the majority of CNS PNET cases. PMID- 19293794 TI - High mesothelin correlates with chemoresistance and poor survival in epithelial ovarian carcinoma. AB - The objective of this paper is to investigate the mesothelin expression level to the clinicopathological features, chemoresponse, and to the outcome of patients with epithelial ovarian carcinoma (EOC). Mesothelin mRNA was detected by real time quantitative reverse-transcription PCR in 139 EOC patients. Clinical characteristics, histopathological items, responses to chemotherapy, progression free survival (PFS), and overall survival (OS) were recorded. Tumours with advanced stages had higher mesothelin than those with early stages. The chemoresistant patients showed significantly higher mesothelin than did chemosensitive patients (2.81 vs 0.43, P<0.001), irrespective of optimal or suboptimal surgery in those with advanced stages. Highly expressed levels of mesothelin were an independent but poor prognostic factor in the PFS (2.03 (1.23 3.37) P=0.006) and OS (3.72 (1.64-8.45), P=0.002) of the 139 EOC patients in multivariate analysis. In addition, patients in advanced stages with highly expressed mesothelin also had significantly worse OS, regardless of whether they had undergone optimal (13.85 (1.76-125.60), P=0.013) or suboptimal (4.47 (1.83 10.88), P=0.001) debulking surgery in multivariate analysis. Out results provide new evidence that mesothelin expression is associated with chemoresistance and with shorter disease-free survival and worse OS of patients with EOC. PMID- 19293795 TI - Extracellular, cell-permeable survivin inhibits apoptosis while promoting proliferative and metastatic potential. AB - The tumour microenvironment is believed to be involved in development, growth, metastasis, and therapy resistance of many cancers. Here we show survivin, a member of the inhibitor of apoptosis protein (IAP) family, implicated in apoptosis inhibition and the regulation of mitosis in cancer cells, exists in a novel extracellular pool in tumour cells. Furthermore, we have constructed stable cell lines that provide the extracellular pool with either wild-type survivin (Surv-WT) or the previously described dominant-negative mutant survivin (Surv T34A), which has proven pro-apoptotic effects in cancer cells but not in normal proliferating cells. Cancer cells grown in conditioned medium (CM) taken from Surv-WT cells absorbed survivin and experienced enhanced protection against genotoxic stresses. These cells also exhibited an increased replicative and metastatic potential, suggesting that survivin in the tumour microenvironment may be directly associated with malignant progression, further supporting survivin's function in tumourigenesis. Alternatively, cancer cells grown in CM taken from the Surv-T34A cells began to apoptose through a caspase-2- and caspase-9 dependent pathway that was further enhanced by the addition of other chemo- and radiotherapeutic modalities. Together our findings suggest a novel microenvironmental function for survivin in the control of cancer aggressiveness and spread, and should result in the genesis of additional cancer treatment modalities. PMID- 19293796 TI - Mean sojourn time, overdiagnosis, and reduction in advanced stage prostate cancer due to screening with PSA: implications of sojourn time on screening. AB - This study aimed to assess the mean sojourn time (MST) of prostate cancer, to estimate the probability of overdiagnosis, and to predict the potential reduction in advanced stage disease due to screening with PSA. The MST of prostate cancer was derived from detection rates at PSA prevalence testing in 43,842 men, aged 50 69 years, as part of the ProtecT study, from the incidence of non-screen-detected cases obtained from the English population-based cancer registry database, and from PSA sensitivity obtained from the medical literature. The relative reduction in advanced stage disease was derived from the expected and observed incidences of advanced stage prostate cancer. The age-specific MST for men aged 50-59 and 60 69 years were 11.3 and 12.6 years, respectively. Overdiagnosis estimates increased with age; 10-31% of the PSA-detected cases were estimated to be overdiagnosed. An interscreening interval of 2 years was predicted to result in 37 and 63% reduction in advanced stage disease in men 65-69 and 50-54 years, respectively. If the overdiagnosed cases were excluded, the estimated reductions were 9 and 54%, respectively. Thus, the benefit of screening in reducing advanced stage disease is limited by overdiagnosis, which is greater in older men. PMID- 19293797 TI - Cetuximab plus gemcitabine/oxaliplatin (GEMOXCET) in first-line metastatic pancreatic cancer: a multicentre phase II study. AB - Targeting the epidermal growth factor receptor pathway in pancreatic cancer seems to be an attractive therapeutic approach. This study assessed the efficacy of cetuximab plus the combination of gemcitabine/oxaliplatin in metastatic pancreatic cancer. Eligible subjects had histological or cytological diagnosis of metastatic pancreatic adenocarcinoma. The primary end point was response according to RECIST. Patients received cetuximab 400 mg m(-2) at first infusion followed by weekly 250 mg m(-2) combined with gemcitabine 1000 mg m(-2) as a 100 min infusion on day 1 and oxaliplatin 100 mg m(-2) as a 2-h infusion on day 2 every 2 weeks. Between January 2005 and August 2006, a total of 64 patients (22 women (34%), 42 men (66%); median age 64 years (range 31-78)) were enrolled at seven study centres. On October 2007, a total of 17 patients were alive. Sixty two patients were evaluable for baseline and 61 for assessment of response to treatment in an intention-to-treat analysis. Six patients had an incomplete drug combination within the first cycle of the treatment plan (n=4 hypersensitivity reactions to the first cetuximab infusion, n=2 refused to continue therapy). Reported grade 3/4 toxicities (% of patients) were leukopaenia 15%, anaemia 8%, thrombocytopaenia 10%, diarrhoea 7%, nausea 18%, infection 18% and allergy 7%. Cetuximab-attributable skin reactions occurred as follows: grade 0: 20%, grade 1: 41%, grade 2: 30% and grade 3: 10%. The intention-to-treat analysis of 61 evaluable patients showed an overall response rate of 33%, including 1 (2%) complete and 19 (31%) partial remissions. There were 31% patients with stable and 36% with progressive disease or discontinuation of the therapy before re-staging. The presence of a grade 2 or higher skin rash was associated with a higher likelihood of achieving objective response. Median time to progression was 118 days, with a median overall survival of 213 days. A clinical benefit response was noted in 24 of the evaluable 61 patients (39%). The addition of cetuximab to the combination of gemcitabine and oxaliplatin is well tolerated but does not increase response or survival in patients with metastatic pancreatic cancer. PMID- 19293798 TI - The BAG-1 cochaperone is a negative regulator of p73-dependent transcription. AB - High-level expression of Bcl-2 associated athanogene (BAG-1) protects cancer cells from stress-induced cell death and growth inhibition. These protective effects of BAG-1 are dependent on interactions with the HSC70 and HSP70 chaperones. However, the key stress-response molecules that are regulated by a BAG-1/chaperone mechanism have not been identified. In this study, we investigated the effects of BAG-1 overexpression on the function of p53 family proteins, p53, p63 and p73. Overexpression of BAG-1 isoforms interfered with the transactivating activity of p73 and p63, but had modest and variable effects on p53-dependent transcription. p73 and BAG-1 interacted in intact cells and overexpression of BAG-1 decreased the expression of p73. siRNA-mediated ablation of endogenous BAG-1 increased the activity of a p73-responsive promoter and this was reversed by knock-down of p73. The ability of BAG-1 to modulate p73 activity and expression, and to interact with p73 were dependent on amino acid residues required for the interaction of BAG-1 with HSC70 and HSP70. These results show that BAG-1 inhibits the transactivating functions of p73 and provide new insight into the mechanisms that control the expression of p73. Inhibition of p73 function may be one mechanism that contributes to the pro-survival activity of BAG-1. PMID- 19293799 TI - Characteristics and outcomes of patients with advanced non-small-cell lung cancer who declined to participate in randomised clinical chemotherapy trials. AB - There are inadequate data on the outcomes of patients who declined to participate in randomised clinical trials as compared with those of participants. We retrospectively reviewed the patient characteristics and treatment outcomes of both participants and non-participants in the two randomised trials for chemotherapy-naive advanced non-small-cell lung cancer. Trial 1 compared four platinum-based combination regimens. Trial 2 compared two sequences of carboplatin plus paclitaxel and gefitinib therapies. Nineteen of 119 (16%) and 153 (37%) patients declined to participate in Trials 1 and 2, respectively. Among the background patient characteristics, the only variable associated with trial participation or declining was the patients' attending physicians (P<0.001). Important differences were not observed in the clinical outcomes between participants and non-participants, for whom the response rates were 30.6 vs 34.2% and the median survival times were 489 vs 461 days, respectively. The hazard ratio for overall survival, adjusted for other confounding variables, was 0.965 (95% confidence interval: 0.73-1.28). In conclusion, there was no evidence to suggest any difference in the characteristics and clinical outcomes between participants and non-participants. Trial designs and the doctor-patient relationship may have an impact on the patient accrual to randomised trials. PMID- 19293800 TI - Breast density and outcome of mammography screening: a cohort study. AB - The purpose of this study was to investigate the effect of breast density on breast cancer (BC) mortality in a mammography screening programme. The cohort included 48 052 women participating in mammography screening in Copenhagen, Denmark, where biennial screening is offered to women aged 50-69 years. We collected information for the years 1991-2001 on screening outcome, incident BCs (screen-, interval-, and later detected), and BC deaths. Breast density was dichotomised into fatty (F) and mixed/dense (M/D) breasts. Screening sensitivity was measured as the odds ratio of interval versus screen-detected cancer for dense versus F breasts. Poisson regression was used to estimate the ratios for BC incidence, case fatality, and mortality between women with M/D and F breasts. For women with M/D breasts, the odds ratio of an interval cancer was 1.62 (95% confidence interval, CI, 1.14-2.30), and the age-adjusted rate ratios were 2.45 (95% CI 2.14-2.81) for BC incidence, 0.60 (95% CI 0.43-0.84) for case fatality, and 1.78 (95% CI 1.17-2.72) for BC mortality. The study shows that BC in women with M/D breasts is more frequent, but on average less severe, than in women with F breasts. PMID- 19293801 TI - High cyclin B1 expression is associated with poor survival in breast cancer. AB - Cyclin B1 regulates the G(2)-M transition of the cell cycle. Cyclin B1 expression is higher in premalignant and malignant than normal breast lesions. Correlation of cyclin B1 expression with other histopathological variables and prognostic role in breast cancer are not fully understood. Traditionally used prognostic criteria identify large subset of patients to receive adjuvant chemotherapy and to be exposed to adverse effects. A reliable and simple method helping prognostic evaluation in breast cancer is needed. We analysed cyclin B1 expression on 1348 invasive breast cancers and studied correlations with other histopathological variables and survival. High cyclin B1 correlated with high tumour grade, large tumour size and positive nodal status, oestrogen and progesterone receptor negativity, positive HER2 and p53 status, young age at diagnosis, and high cyclin E, cyclin A and Ki67 expression. Among patients not given adjuvant chemotherapy high cyclin B1 was a strong predictor of shorter overall and metastasis-free survival (RR 3.74, P<0.0005 and RR 3.51, P<0.0005, respectively), and remained as an independent prognostic factor also in multivariate analysis (RR 1.80, P=0.04 and RR 2.31, P=0.02, respectively). This study suggests high cyclin B1 associates with aggressive phenotype and is an independent prognostic factor in breast cancer. PMID- 19293802 TI - Persistence of HPV infection and risk of high-grade cervical intraepithelial neoplasia in a cohort of Colombian women. AB - Little is known about the dynamics of human papillomavirus (HPV) infection and subsequent development of high-grade cervical intraepithelial neoplasia (CIN2/3), particularly in women >30 years of age. This information is needed to assess the impact of HPV vaccines and consider new screening strategies. A cohort of 1728 women 15-85 years old with normal cytology at baseline was followed every 6 months for an average of 9 years. Women with squamous intraepithelial lesions were referred for biopsy and treatment. The Kaplan-Meier method was used to estimate the median duration of infection and Cox regression analysis was undertaken to assess determinants of clearance and risk of CIN2/3 associated with HPV persistence. No difference in the likelihood of clearance was observed by HPV type or woman's age, with the exception of lower clearance for HPV16 infection in women under 30 years of age. Viral load was inversely associated with clearance. In conclusion, viral load is the main determinant of persistence, and persistence of HPV16 infections carry a higher risk of CIN2/3. PMID- 19293803 TI - Differing deregulation of EGFR and downstream proteins in primary colorectal cancer and related metastatic sites may be clinically relevant. AB - Cetuximab and panitumumab efficacy in metastatic colorectal cancer (mCRC) may be influenced by EGFR gene status and/or deregulation of its downstream signalling proteins detected in primary tumour. However, metastasis might have different molecular patterns with respect to primary tumour, possibly affecting the prediction of EGFR-targeted therapy efficacy. We analysed primary tumour and metastasis in 38 mCRC patients. Twelve cases were cetuximab/panitumumab treated. EGFR gene status and protein expression were investigated through fluorescent in situ hybridisation and immunohistochemistry (IHC), K-Ras/BRAF mutations by sequencing and PTEN expression by IHC. We observed EGFR gene deregulation in 25 out of 36 primary tumours and 29 out of 36 metastases, K-Ras mutations in 16 out of 37 cancers and in 15 out of 37 metastases, BRAF mutations in 2 out of 36 cancers and 2 out of 36 metastases and PTEN loss in 8 out of 38 cancers and 12 out of 38 metastases. For the first time in literature, we show that primary colorectal cancer and paired metastasis may exhibit difference with respect to EGFR pathway deregulation mechanisms possibly implying a different response to cetuximab or panitumumab treatment. The investigation of treated patients confirms this hypothesis. We therefore suggest that the analysis of metastatic lesion should be considered in patient management as well as in designing future clinical trials aimed to investigate the effect of anti-EGFR monoclonal antibodies in the treatment of mCRC. PMID- 19293804 TI - Incidence and predictors of treatment-related mortality in paediatric acute leukaemia in El Salvador. AB - Survival rates among children with leukaemia in low-income countries are lower than those in high-income countries. This has been attributed in part to higher treatment-related mortality (TRM). We examined the demographics, treatment, and outcomes of paediatric patients in El Salvador with acute lymphoblastic leukaemia (ALL) or acute myeloid leukaemia (AML) to determine the incidence, causes, and risk factors for TRM. Two trained data managers collected data prospectively; no patients were excluded. Biological, socioeconomic and nutritional predictors were examined. A total of 469 patients with ALL and 78 patients with AML were included. The 2-year cumulative incidence of TRM was significantly higher among children with AML (35.4+/-6.4%) than those with ALL (12.5+/-1.7%; P<0.0001). However, the proportion of deaths attributable to the toxicity of treatment did not differ significantly between AML (25/47, 53.2%) and ALL (55/107, 51.4%; P=0.98). Among children with ALL, low monthly income (P=0.04) and low parental education (P=0.02) significantly increased the risk of TRM. Among children with AML, biological, socioeconomic, and nutritional variables were not associated with TRM. In this low-income country, toxic death significantly contributes to mortality in both ALL and AML. A better understanding of the effect of socioeconomic status on TRM may suggest specific strategies for patients with ALL. PMID- 19293805 TI - Expression of Mcm2, geminin and Ki67 in normal oral mucosa, oral epithelial dysplasias and their corresponding squamous-cell carcinomas. AB - Proteins necessary for the normal regulation of the cell cycle include minichromosome maintenance protein 2 (Mcm2) and geminin. These are overexpressed in several premalignant and malignant tumours. The Mcm2/Ki67 ratio can be used to estimate the population of cells that are in early G(1) (licensed to proliferate), and the geminin/Ki67 ratio can determine the relative length of G(1). A high ratio indicates a short G(1) and a high rate of cell proliferation. Mcm2 and geminin have been scarcely explored in oral epithelial dysplasia (OED) and oral squamous-cell carcinoma (OSCC). The purpose of this study was to identify the expression pattern of Mcm2, Ki67 and geminin in normal oral mucosa (NOM), OED and their subsequent OSCC, to determine if expression could help predict the prognosis of OED. Paraffin sections of 41 OED cases that progressed to carcinoma, 40 OED without malignant progression, 38 OSCC and 15 NOM were immunostained with antibodies against Mcm2, geminin and Ki67. Labelling indices (LIs) increased progressively from NOM, OED and OSCC (Mcm2, P<0.001; geminin, P<0.001 and Ki67, P<0.001). In all the OED cases (n=81) the levels of expression of Mcm2 (LI, 73.6), geminin (LI, 24.4) and Ki67 (LI, 44.5) were elevated indicating a constant cell-cycle re-entry. When the OED groups were compared, Mcm2 protein expression was higher in the OED with malignant progression (P=0.04), likewise there was a significant increase in the Mcm2/Ki67 and geminin/Ki67 ratios (P=0.04 and 0.02 respectively). Mcm2 and geminin proteins seem to be novel biomarkers of growth and may be useful prognostic tools for OED. PMID- 19293806 TI - Homozygous CDA*3 is a major cause of life-threatening toxicities in gemcitabine treated Japanese cancer patients. AB - Among 242 Japanese pancreatic cancer patients, three patients (1.2%) encountered life-threatening toxicities, including myelosuppression, after gemcitabine-based chemotherapies. Two of them carried homozygous CDA*3 (CDA208G>A [Ala70Thr]), and showed extremely low plasma cytidine deaminase activity and gemcitabine clearance. Our results suggest that homozygous *3 is a major factor causing gemcitabine-mediated severe adverse reactions among the Japanese population. PMID- 19293807 TI - Ki-67 and outcome in clinically localised prostate cancer: analysis of conservatively treated prostate cancer patients from the Trans-Atlantic Prostate Group study. AB - Treatment decisions after diagnosis of clinically localised prostate cancer are difficult due to variability in tumour behaviour. We therefore examined one of the most promising biomarkers in prostate cancer, Ki-67, in a cohort of 808 patients diagnosed with prostate cancer between 1990 and 1996 and treated conservatively. Ki-67 expression was assessed immunohistochemically, in two laboratories, by two different scoring methods and the results compared with cancer-specific and overall survival. The power of the biomarker was compared with Gleason score and initial serum prostate-specific antigen (PSA). Both methods showed that Ki-67 provided additional prognostic information beyond that available from Gleason score and PSA: for the semi-quantitative method, Deltachi(2) (1 d.f.)=24.6 (P<0.0001), overall survival chi(2)=20.5 (P<0.0001), and for the quantitative method, Deltachi(2) (1 d.f.)=15.1 (P=0.0001), overall survival chi(2)=10.85 (P=0.001). Ki-67 is a powerful biomarker in localised prostate cancer and adds to a model predicting the need for radical or conservative therapy. As it is already in widespread use in routine pathology, it is confirmed as the most promising biomarker to be applied into routine practice. PMID- 19293808 TI - A tool to balance benefit and harm when deciding about adjuvant therapy. AB - Adjuvant therapy aims to prevent outgrowth of residual disease but can induce serious side effects. Weighing conflicting treatment effects and communicating this information with patients is not elementary. This study presents a scheme balancing benefit and harm of adjuvant therapy vs no adjuvant therapy. It is illustrated by the available evidence on adjuvant pelvic external beam radiotherapy (RT) for intermediate-risk stage I endometrial carcinoma patients. The scheme comprises five outcome possibilities of adjuvant therapy: patients who benefit from adjuvant therapy (some at the cost of complications) vs those who neither benefit nor contract complications, those who do not benefit but contract severe complications, or those who die. Using absolute risk differences, a fictive cohort of 1000 patients receiving adjuvant RT is categorised. Three large randomised clinical trials were included. Recurrences will be prevented by adjuvant RT in 60 patients, a majority of 908 patients will neither benefit nor suffer severe radiation-induced harm but 28 patients will suffer severe complications due to adjuvant RT and an expected four patients will die. This scheme readily summarises the different possible treatment outcomes and can be of practical value for clinicians and patients in decision making about adjuvant therapies. PMID- 19293809 TI - Radiosensitisation of U87MG brain tumours by anti-epidermal growth factor receptor monoclonal antibodies. AB - As epidermal growth factor receptor (EGFR) has been reported to be a radiation response modulator, HER inhibitors are regarded to act as potential radiosensitisers. Our study examined the role of nimotuzumab and cetuximab both, the two monoclonal antibodies (mAbs) to EGFR, as radiosensitisers in a murine glioma model in vivo. Co-administration of both the antibodies with radiation increased the radiosensitivity of U87MG, resulting in a significant delay of subcutaneous (s.c.) tumour growth. Furthermore, the addition of antibodies to the radiation decreased brain tumour sizes and is inhibited by 40-80% the increased tumour cell invasion provoked by radiotherapy, although promoted tumour cell apoptosis. Whereas nimotuzumab led to a reduction in the size of tumour blood vessels and proliferating cells in s.c. tumours, cetuximab had no significant antiangiogenic nor antiproliferative activity. In contrast, cetuximab induced a more marked inhibition of EGFR downstream signalling compared with nimotuzumab. Moreover, both antibodies reduced the total number of radioresistant CD133+ cancer stem cells (CSCs). These results were encouraging, and showed the superiority of combined treatment of mAbs to EGFR and radiation over each single therapy against glioblastoma multiforme (GBM), confirming the role of these drugs as radiosensitisers in human GBM. In addition, we first showed the ability of mAb specifics against EGFR to target radioresistant glioma CSC, supporting the potential use in patients. PMID- 19293810 TI - External validation of serum hCG cutoff levels for prediction of resistance to single-agent chemotherapy in patients with persistent trophoblastic disease. AB - Van Trommel et al have previously shown that serum human chorionic gonadotropin (hCG) cutoff levels can provide early prediction of resistance to first-line methotrexate (MTX) in patients with persistent trophoblastic disease (PTD). In this study, we validate this approach of prediction of resistance to single-agent chemotherapy in an independent and larger cohort of PTD patients using a different hCG assay. Receiver operating characteristics (ROC) curves were constructed to determine hCG cutoff levels and sensitivity between patients cured on single-agent chemotherapy (control group) and patients requiring change to combination chemotherapy (study group). Receiver operating characteristics analysis identified an hCG cutoff value of 737 IU l(-1) that enabled us to predict the subsequent development of single-agent chemotherapy resistance in 52% of patients before their fourth MTX course at 97.5% specificity. This would have enabled an earlier switch to combination chemotherapy reducing the MTX exposure by an average of 2.5 courses. The present findings confirm that serum hCG cutoff levels predict resistance to single-agent therapy earlier than traditional methods. Change to combination chemotherapy should be considered for patients whose serum hCG levels exceed these hCG cutoff values. For patients not exceeding the hCG cutoff levels, static or rising hCG levels should still be included in the criteria for change of chemotherapy. PMID- 19293811 TI - Detection of K-Ras mutations in tumour samples of patients with non-small cell lung cancer using PNA-mediated PCR clamping. AB - Non-small cell lung cancers (NSCLC), in particular adenocarcinoma, are often mixed with normal cells. Therefore, low sensitivity of direct sequencing used for K-Ras mutation analysis could be inadequate in some cases. Our study focused on the possibility to increase the detection of K-Ras mutations in cases of low tumour cellularity. Besides direct sequencing, we used wild-type hybridisation probes and peptide-nucleic-acid (PNA)-mediated PCR clamping to detect mutations at codons 12 and 13, in 114 routine consecutive NSCLC frozen surgical tumours untreated by targeted drugs. The sensitivity of the analysis without or with PNA was 10 and 1% of tumour DNA, respectively. Direct sequencing revealed K-Ras mutations in 11 out of 114 tumours (10%). Using PNA-mediated PCR clamping, 10 additional cases of K-Ras mutations were detected (21 out of 114, 18%, P<0.005), among which five in samples with low tumour cellularity. In adenocarcinoma, K-Ras mutation frequency increased from 7 out of 55 (13%) by direct sequencing to 15 out of 55 (27%) by clamped-PCR (P<0.005). K-Ras mutations detected by these sensitive techniques lost its prognostic value. In conclusion, a rapid and sensitive PCR-clamping test avoiding macro or micro dissection could be proposed in routine analysis especially for NSCLC samples with low percentage of tumour cells such as bronchial biopsies or after neoadjuvant chemotherapy. PMID- 19293812 TI - MicroRNA deregulation and pathway alterations in nasopharyngeal carcinoma. AB - MicroRNAs (miRNAs) are a family of small non-coding RNA molecules of about 20-23 nucleotides in length, which negatively regulate protein-coding genes at post transcriptional level. Using a stem-loop real-time-PCR method, we quantified the expression levels of 270 human miRNAs in 13 nasopharyngeal carcinoma (NPC) samples and 9 adjacent normal tissues, and identified 35 miRNAs whose expression levels were significantly altered in NPC samples. Several known oncogenic miRNAs, including miR-17-92 cluster and miR-155, are among the miRNAs upregulated in NPC. Tumour suppressive miRNAs, including miR-34 family, miR-143, and miR-145, are significantly downregulated in NPC. To explore the roles of these dysregulated miRNAs in the pathogenesis of NPC, a computational analysis was performed to predict the pathways collectively targeted by the 22 significantly downregulated miRNAs. Several biological pathways that are well characterised in cancer are significantly targeted by the downregulated miRNAs. These pathways include TGF Wnt pathways, G1-S cell cycle progression, VEGF signalling pathway, apoptosis and survival pathways, and IP3 signalling pathways. Expression levels of several predicted target genes in G1-S progression and VEGF signalling pathways were elevated in NPC tissues and showed inverse correlation with the down-modulated miRNAs. These results indicate that these downregulated miRNAs coordinately regulate several oncogenic pathways in NPC. PMID- 19293816 TI - Irinotecan toxicity: genes or intestinal microflora? PMID- 19293819 TI - The evolving understanding of COPI vesicle formation. AB - The coat protein I (COPI) complex is considered to be one of the best characterized coat complexes. Studies on how it functions in vesicle formation have provided seminal contributions to the general paradigm in vesicular transport that the ADP-ribosylation factor (ARF) small GTPases are key regulators of coat complexes. Here, we discuss emerging evidence that suggests the need to revise some long-held views on how COPI vesicle formation is achieved. PMID- 19293820 TI - Human genetic variation and its contribution to complex traits. AB - The last few years have seen extensive efforts to catalogue human genetic variation and correlate it with phenotypic differences. Most common SNPs have now been assessed in genome-wide studies for statistical associations with many complex traits, including many important common diseases. Although these studies have provided new biological insights, only a limited amount of the heritable component of any complex trait has been identified and it remains a challenge to elucidate the functional link between associated variants and phenotypic traits. Technological advances, such as the ability to detect rare and structural variants, and a clear understanding of the challenges in linking different types of variation with phenotype, will be essential for future progress. PMID- 19293821 TI - Monocyte subtypes predict clinical course and prognosis in human stroke. AB - The number of circulating monocytes increases after stroke. In this study, we assessed the time course and phenotype of monocyte subsets and their relationship with the clinical course and outcome in 46 consecutive stroke patients and 13 age matched controls. The proportion of the most abundant 'classical' CD14(high)CD16- monocytes did not change after stroke, whereas that of CD14(high)CD16+ monocytes increased and CD14(dim)CD16+ monocytes decreased. CD14(high)CD16+ monocytes had the highest expression of TLR2, HLA-DR and the angiogenic marker, Tie-2; CD14(dim)CD16+ monocytes had the highest expression of costimulatory CD86 and adhesion molecule CD49d. Platelet-monocyte interactions were highest in CD14(high)CD16- monocytes and lowest in CD14(dim)CD16+ monocytes. In adjusted models, 1/CD14(high)CD16- monocytes were associated with poor outcome (OR: 1.38), higher mortality (OR: 1.40) and early clinical worsening (OR: 1.29); 2/CD14(high)CD16+ monocytes were inversely related to mortality (OR: 0.32); and 3/CD14(dim)CD16+ monocytes were inversely related to poor outcome (OR: 0.74) and infarction size (r=-0.45; P=0.02). These results illustrate that the predominant monocyte subtype conveys harmful effects after stroke, which include stronger interaction with platelets. Alternatively, rarer subpopulations of monocytes are beneficial with a phenotype that could promote tissue repair and angiogenesis. Therefore, monitoring of monocyte subtypes may emerge as a useful tool at the bedside for stroke patients. PMID- 19293822 TI - The importance of latency in the focality of perfusion and oxygenation changes associated with triggered afterdischarges in human cortex. AB - The spatiotemporal dynamics of neurovascular coupling during epilepsy are not well understood, and there are little data from studies of the human brain. We investigated changes in total hemoglobin (Hbt) and hemoglobin oxygenation in patients undergoing epilepsy surgery with intraoperative intrinsic optical spectroscopy (IOS) during triggered afterdischarges (ADs). We found an early (approximately 0.5 secs) focal dip in hemoglobin oxygenation, arising precisely in the stimulated gyrus that lasted for 11.5+/-10.0 secs, approximately the length of the AD (10.4+/-4.4 secs). A later oxygen overshoot and increase in blood volume occurred in the adjacent surrounding gyri. After a significant delay (approximately 20 to 30 secs), the overshoot and blood volume signal became extremely focal to the area of the onset of the AD. A smaller very late undershoot, the last phase of the 'triphasic' response, was also identified, although localization was inconsistent. In this study, we show that a 'late focal overshoot' and late Hbt signal may be extremely useful, in addition to the early dip, for the localization of seizure onset. It is likely that a separate mechanism underlies the persistent focal increase in cerebral blood volume after a long-duration cortical stimulation, compared with the nonspecific mechanism that causes the initial increase in cerebral blood flow. PMID- 19293823 TI - Critical closing pressure: comparison of three methods. AB - Critical closing pressure (CCP) is an arterial pressure threshold below which small arterial vessels collapse. Our aim was to compare different methods to estimate CCP in the cerebrovascular circulation using the relationships between transcranial Doppler flow velocity (FV), laser-Doppler flux (LDF), and arterial blood pressure (ABP). A total of 116 experiments in rabbits were analyzed retrospectively. At the end of each recording, cardiac arrest (CA) was induced. Arterial blood pressure in femoral artery, basilar artery FV, cortical blood LDF, intracranial pressure (ICP) was recorded. Critical closing pressure was estimated using linear regression between decreasing mean ABP values, FV, and LDF during CA. In addition, CCP was calculated from FV waveform just before CA. The correlation between CCP evaluated using LDF and FV during CA was 0.98 (P<0.0001). The correlation between CCP measured during CA and CCP estimated from the transcranial Doppler ultrasonography (TCD) waveform was weaker (R=0.39; P<0.001), with CCP calculated from waveform being significantly greater than CCP from CA (median difference 9 mm Hg; P<0.003). Critical closing pressures obtained from FV waveform and CA correlated with mean ICP before CA (R=0.40; P=0.001). In conclusion strong correlation exists between CCP values obtained by means of FV and LDF during cardiac arrest. However, predictions of CCP using TCD waveform analysis show substantial differences from values of CCP recorded during cardiac arrest. PMID- 19293824 TI - Correction for the effect of rising plasma glucose levels on quantification of MR(glc) with FDG-PET. AB - Positron emission tomography (PET) using the tracer [18F]-fluorodeoxyglucose (FDG) is commonly used for measuring metabolic rate of glucose (MR(glc)) in the human brain. Conventional PET methods (e.g., the Patlak method) for quantifying MR(glc) assume the tissue transport and phosphorylation mechanisms to be in steady state during FDG uptake. As FDG and glucose use the same transporters and phosphorylation enzymes, changing blood glucose levels can change the rates of FDG transport and phosphorylation. Compartmental models were used to simulate the effect of rising arterial glucose, from normal to hyperglycemic levels on FDG uptake for a typical PET protocol. The subsequent errors on the values of MR(glc) calculated using the Patlak method were investigated, and a correction scheme based on measured arterial glucose concentration (G(p)) was evaluated. Typically, with a 40% rise in G(p) over the duration of the PET study, the true MR(glc) varied by only 1%; however, the Patlak method overestimated MR(glc) by 15%. The application of the correction reduced this error to approximately 2%. In general, the application of the correction resulted in values of MR(glc) consistently significantly closer to the true steady state calculation of MR(glc) independently of changes to the parameters defining the model. PMID- 19293825 TI - Early metabolic characteristics of lesion and nonlesion tissue after head injury. AB - We defined lesion and structurally normal regions using magnetic resonance imaging at follow-up in patients recovering from head injury. Early metabolic characteristics in these regions of interest (ROIs) were compared with physiology in healthy volunteers. Fourteen patients with severe head injury had positron emission tomography within 72 h, and magnetic resonance imaging at 3 to 18 months after injury. Cerebral blood flow (CBF), oxygen utilization (CMRO(2)), and oxygen extraction fraction (OEF) were all lower in lesion ROIs, compared with nonlesion and control ROIs (P<0.001); however, there was substantial overlap in physiology. Control ROIs showed close coupling among CBF, blood volume (CBV), and CMRO(2), whereas relationships within lesion and nonlesion ROIs were abnormal. The relationship between CBF and CMRO(2) generally remained coupled but the slope was reduced; that for CBF and OEF was variable; whereas that between CBF and CBV was highly variable. There was considerable heterogeneity between and within patients. Although irreversibly damaged tissue is characterized by marked derangements in physiology, a more detailed analysis shows acute changes in physiology and physiologic relationships within regions of the brain that appear structurally normal at follow-up. Such pathophysiological derangements may result in selective neuronal loss and impact on functional outcome. PMID- 19293826 TI - Force- and length-dependent catastrophe activities explain interphase microtubule organization in fission yeast. AB - The cytoskeleton is essential for the maintenance of cell morphology in eukaryotes. In fission yeast, for example, polarized growth sites are organized by actin, whereas microtubules (MTs) acting upstream control where growth occurs. Growth is limited to the cell poles when MTs undergo catastrophes there and not elsewhere on the cortex. Here, we report that the modulation of MT dynamics by forces as observed in vitro can quantitatively explain the localization of MT catastrophes in Schizosaccharomyces pombe. However, we found that it is necessary to add length-dependent catastrophe rates to make the model fully consistent with other previously measured traits of MTs. We explain the measured statistical distribution of MT-cortex contact times and re-examine the curling behavior of MTs in unbranched straight tea1Delta cells. Importantly, the model demonstrates that MTs together with associated proteins such as depolymerizing kinesins are, in principle, sufficient to mark the cell poles. PMID- 19293827 TI - Temporal switching and cell-to-cell variability in Ca2+ release activity in mammalian cells. AB - Genetically identical cells in a uniform external environment can exhibit different phenotypes, which are often masked by conventional measurements that average over cell populations. Although most studies on this topic have used microorganisms, differentiated mammalian cells have rarely been explored. Here, we report that only approximately 40% of clonal human embryonic kidney 293 cells respond with an intracellular Ca(2+) increase when ryanodine receptor Ca(2+) release channels in the endoplasmic reticulum are maximally activated by caffeine. On the other hand, the expression levels of ryanodine receptor showed a unimodal distribution. We showed that the difference in the caffeine sensitivity depends on a critical balance between Ca(2+) release and Ca(2+) uptake activities, which is amplified by the regenerative nature of the Ca(2+) release mechanism. Furthermore, individual cells switched between the caffeine-sensitive and caffeine-insensitive states with an average transition time of approximately 65 h, suggestive of temporal fluctuation in endogenous protein expression levels associated with caffeine response. These results suggest the significance of regenerative mechanisms that amplify protein expression noise and induce cell-to cell phenotypic variation in mammalian cells. PMID- 19293828 TI - Toward accurate reconstruction of functional protein networks. AB - Genome-scale screening studies are gradually accumulating a wealth of data on the putative involvement of hundreds of genes/proteins in various cellular responses or functions. A fundamental challenge is to chart out the protein pathways that underlie these systems. Previous approaches to the problem have either employed a local optimization criterion, aiming to infer each pathway independently, or a global criterion, searching for the overall most parsimonious subnetwork. Here, we study the trade-off between the two approaches and present a new intermediary scheme that provides explicit control over it. We demonstrate its utility in the analysis of the apoptosis network in humans, and the telomere length maintenance (TLM) system in yeast. Our results show that in the majority of real-life cases, the intermediary approach provides the most plausible solutions. We use a new set of perturbation experiments measuring the role of essential genes in telomere length regulation to further study the TLM network. Surprisingly, we find that the proteasome plays an important role in telomere length regulation through its associations with transcription and DNA repair circuits. PMID- 19293829 TI - Personalizing medicine: a systems biology perspective. PMID- 19293831 TI - Toward a comprehensive and quantitative understanding of intracellular microtubule organization. PMID- 19293830 TI - Force- and kinesin-8-dependent effects in the spatial regulation of fission yeast microtubule dynamics. AB - Microtubules (MTs) are central to the organisation of the eukaryotic intracellular space and are involved in the control of cell morphology. For these purposes, MT polymerisation dynamics are tightly regulated. Using automated image analysis software, we investigate the spatial dependence of MT dynamics in interphase fission yeast cells with unprecedented statistical accuracy. We find that MT catastrophe frequencies (switches from polymerisation to depolymerisation) strongly depend on intracellular position. We provide evidence that compressive forces generated by MTs growing against the cell pole locally reduce MT growth velocities and enhance catastrophe frequencies. Furthermore, we find evidence for an MT length-dependent increase in the catastrophe frequency that is mediated by kinesin-8 proteins (Klp5/6). Given the intrinsic susceptibility of MT dynamics to compressive forces and the widespread importance of kinesin-8 proteins, we propose that similar spatial regulation of MT dynamics plays a role in other cell types as well. In addition, our systematic and quantitative data should provide valuable input for (mathematical) models of MT organisation in living cells. PMID- 19293832 TI - Complex topology rather than complex membership is a determinant of protein dosage sensitivity. AB - The 'balance hypothesis' predicts that non-stoichiometric variations in concentrations of proteins participating in complexes should be deleterious. As a corollary, heterozygous deletions and overexpression of protein complex members should have measurable fitness effects. However, genome-wide studies of heterozygous deletions in Saccharomyces cerevisiae and overexpression have been unable to unambiguously relate complex membership to dosage sensitivity. We test the hypothesis that it is not complex membership alone but rather the topology of interactions within a complex that is a predictor of dosage sensitivity. We develop a model that uses the law of mass action to consider how complex formation might be affected by varying protein concentrations given a protein's topological positioning within the complex. Although we find little evidence for combinatorial inhibition of complex formation playing a major role in overexpression phenotypes, consistent with previous results, we show significant correlations between predicted sensitivity of complex formation to protein concentrations and both heterozygous deletion fitness and protein abundance noise levels. Our model suggests a mechanism for dosage sensitivity and provides testable predictions for the effect of alterations in protein abundance noise. PMID- 19293833 TI - Linearization through distortion: a new facet of negative feedback in signalling. PMID- 19293835 TI - Introgression as a likely cause of mtDNA paraphyly in two allopatric skippers (Lepidoptera: Hesperiidae). AB - Gene transfer between species during interspecific hybridization is a widely accepted reality in plants but is considered a relatively rare phenomenon among animals. Here we describe a unique case of mitochondrial DNA (mtDNA) paraphyly in the skipper genus, Erynnis, that involves well-diverged allopatric species. Using molecular evidence from both mitochondrial and nuclear genomes, we found high levels of intraspecific divergence in the mitochondrial genome within E. propertius (over 4% pair-wise sequence divergence) but no such differentiation in the nuclear genome. Sequence comparisons with related Erynnis suggest that past, but recent and infrequent introgression between E. propertius and E. horatius is the most reasonable explanation for the observed pattern of mtDNA paraphyly. This example of putative introgression highlights the complexity of mtDNA evolution and suggests that similar processes could be operating in other taxa that have not been extensively sampled. Our observations reinforce the importance of involving multiple genes with different modes of inheritance in the analysis of population history of congeneric taxa. PMID- 19293836 TI - Inbreeding depression in dioecious populations of the plant Mercurialis annua: comparisons between outcrossed progeny and the progeny of self-fertilized feminized males. AB - Inbreeding depression is a key factor in the maintenance of separate sexes in plants through selection for the avoidance of self-fertilization. However, very little is known about the levels of inbreeding depression in dioecious species, obviously because it is difficult to self-fertilize males or females. We overcame this problem by clonally propagating males from lineages in a dioecious metapopulation of the European annual plant Mercurialis annua, feminizing some of them and crossing the feminized with the unfeminized clones. Using this method, we compared the fitness of selfed vs outcrossed progeny under field conditions in Spain, where this species grows naturally. Multiplicative inbreeding depression (based on seed germination, early and late survival, seed mass and pollen viability) ranged from -0.69 to 0.82, with a mean close to zero. We consider possible explanations for both the low mean and high variance in inbreeding depression in M. annua, and we discuss the implications of our results for the maintenance of dioecy over hermaphroditism. PMID- 19293838 TI - Prevalence data on all Ghent features in a cross-sectional study of 87 adults with proven Marfan syndrome. AB - The prevalence of each single feature in the Ghent criteria in patients with Marfan syndrome (MFS) is not known. To elucidate this, a cross-sectional study of 105 adults with presumed MFS was carried out. All patients were examined by the same group of investigators with standardized and complete assessment of all features in the Ghent criteria. Eighty-seven (83%) fulfilled the criteria in 56 different variants. The most prevalent major criterion in Ghent-positive persons was dural ectasia (91%), followed by major genetic criterion (89%) and ectopic lenses (62 %). In 14 persons (16%), the diagnosis was dependent on the dural findings. In all, 79% fulfilled both major dural and major genetic (positive family history and/or FBN1 mutation) criteria, suggesting that most patients with MFS might be identified by investigating these criteria. A history or finding of ascending aortic disease was present in 46 patients (53%). This low prevalence might partly reflect a high number of diagnosed patients encompassing the whole spectrum of the syndrome. The study confirms the need to examine for the complete set of features in the Ghent criteria to identify all patients with MFS. The majority of persons with MFS might be identified by the combined assessment of dura mater and family history, supplemented with DNA analysis in family-negative cases. The low prevalence of ascending aortic disease might indicate better future prospects in an adult population than those traditionally considered. PMID- 19293839 TI - 16p subtelomeric duplication: a clinically recognizable syndrome. AB - We report on two patients with duplication of the subterminal region of chromosome 16p (dup16p) recognized by fluorescent in situ hybridization (FISH) telomere analysis, presenting with closely overlapping facial features and neurological impairment. Distinct facial anomalies included high forehead, sparse eyebrows, blepharophimosis, short nose, everted upper lip, high-arched palate, wide-spaced teeth, and cupped anteverted ears. Susceptibility to vascular anomalies, in particular pulmonary hypertension and portal cavernoma, was found in one patient. Subtelomeric analysis by FISH demonstrated a de novo duplication of the subtelomeric region of chromosome 16p and a deletion of the subtelomeric region of chromosome 4q in case 1, and duplication of the subtelomeric region of 16p and a deletion of the subtelomeric region of 21q, resulting from malsegregation of a balanced maternal traslocation t(16pter;21qter) in case 2. The extension of duplicated regions measured by array-comparative genome hybridization was about 12 Mb on 16p13.3p13.13 in case 1, and about 8.5 Mb on 16p13.3p13.2 in case 2. In conclusion, we reported a clinically recognizable disorder in two patients with dup16p. Pulmonary hypertension, vascular ring, and manifestations of vascular disruption, as terminal hypoplasia of hands and aplasia cutis, have been previously described in association with dup16p. Thus, susceptibility to pulmonary vascular disease and other vascular anomalies can be a feature of dup16p, suggesting that this subtelomeric region in some respect could be related to vascular anomalies. PMID- 19293837 TI - Replication analysis identifies TYK2 as a multiple sclerosis susceptibility factor. AB - In a recent genome-wide association study (GWAS) based on 12,374 non-synonymous single nucleotide polymorphisms we identified a number of candidate multiple sclerosis susceptibility genes. Here, we describe the extended analysis of 17 of these loci undertaken using an additional 4234 patients, 2983 controls and 2053 trio families. In the final analysis combining all available data, we found that evidence for association was substantially increased for one of the 17 loci, rs34536443 from the tyrosine kinase 2 (TYK2) gene (P=2.7 x 10(-6), odds ratio=1.32 (1.17-1.47)). This single nucleotide polymorphism results in an amino acid substitution (proline to alanine) in the kinase domain of TYK2, which is predicted to influence the levels of phosphorylation and therefore activity of the protein and so is likely to have a functional role in multiple sclerosis. PMID- 19293840 TI - The role of sarcomere gene mutations in patients with idiopathic dilated cardiomyopathy. AB - We investigated a Danish cohort of 31 unrelated patients with idiopathic dilated cardiomyopathy (IDC), to assess the role that mutations in sarcomere protein genes play in IDC. Patients were genetically screened by capillary electrophoresis single strand conformation polymorphism and subsequently by bidirectional DNA sequencing of conformers in the coding regions of MYH7, MYBPC3, TPM1, ACTC, MYL2, MYL3, TNNT2, CSRP3 and TNNI3. Eight probands carried disease associated genetic variants (26%). In MYH7, three novel mutations were found; in MYBPC3, one novel variant and two known mutations were found; and in TNNT2, a known mutation was found. One proband was double heterozygous. We find evidence of phenotypic plasticity: three mutations described earlier as HCM causing were found in four cases of IDC, with no history of a hypertrophic phase. Furthermore, one pedigree presented with several cases of classic DCM as well as one case with left ventricular non-compaction. Disease-causing sarcomere gene mutations were found in about one-quarter of IDC patients, and seem to play an important role in the causation of the disease. The genetics is as complex as seen in HCM. Thus, our data suggest that a genetic work-up should include screening of the most prominent sarcomere genes even in the absence of a family history of the disease. PMID- 19293841 TI - The interaction index, a novel information-theoretic metric for prioritizing interacting genetic variations and environmental factors. AB - We developed an information-theoretic metric called the Interaction Index for prioritizing genetic variations and environmental variables for follow-up in detailed sequencing studies. The Interaction Index was found to be effective for prioritizing the genetic and environmental variables involved in GEI for a diverse range of simulated data sets. The metric was also evaluated for a 103-SNP Crohn's disease dataset and a simulated data set containing 9187 SNPs and multiple covariates that was modeled on a rheumatoid arthritis data set. Our results demonstrate that the Interaction Index algorithm is effective and efficient for prioritizing interacting variables for a diverse range of epidemiologic data sets containing complex combinations of direct effects, multiple GGI and GEI. PMID- 19293842 TI - Clinical features and molecular modelling of novel MPZ mutations in demyelinating and axonal neuropathies. AB - Mutations in the myelin protein zero (MPZ) gene have been associated with different Charcot-Marie-Tooth disease (CMT) phenotypes, including classical demyelinating CMT1B and the axonal form of the disease (CMT2). The MPZ role in the pathogenesis of both demyelinating and axonal inherited neuropathies was evaluated in the Italian population by screening a cohort of 214 patients with CMT1 or CMT2. A MPZ mutation frequency of 7.9% in demyelinating cases and of 4.8% in axonal cases was observed. In the total cohort (264 patients), including those with mutations in other genes, a mutation frequency of 5.8% (7/121) in demyelinating cases and 4.2% (6/143) in axonal cases was found. Three novel MPZ mutations, two missense (p.Ser111Cys, p.Thr124Ala) and one frameshift (p.Tyr145fs) were found, and a molecular modelling approach was used to test the effects of these mutations on the protein structure. Electrostatic distribution changes within the protein, caused by the amino acid substitution, fit in with phenotypes presented by patients herein described. Our findings suggest that the clinical features associated with MPZ mutations depend partly on the nature of amino acid change and that molecular modelling may provide useful support, based on effects on secondary and tertiary protein structure, to predict the phenotype associated with MPZ mutations. PMID- 19293843 TI - Identification of the minimal combination of clinical features in probands for efficient mutation detection in the FBN1 gene. AB - Mutations identified in the fibrillin-1 (FBN1) gene have been associated with Marfan syndrome (MFS). Molecular analysis of the gene is classically performed in probands with MFS to offer diagnosis for at-risk relatives and in children highly suspected of MFS. However, FBN1 gene mutations are found in an ill-defined group of diseases termed 'type I fibrillinopathies', which are associated with an increased risk of aortic dilatation and dissection. Thus, there is growing awareness of the need to identify these non-MFS probands, for which FBN1 gene screening should be performed. To answer this need we compiled the molecular data obtained from the screening of the FBN1 gene in 586 probands, which had been addressed to our laboratory for molecular diagnosis. In this group, the efficacy of FBN1 gene screening was high in classical MFS probands (72.5%,), low (58%) in those referred for incomplete MFS and only slight (14.3%) for patients referred as possible MFS. Using recursive partitioning, we found that the best predictor of the identification of a mutation in the FBN1 gene was the presence of features in at least three organ systems, combining one major, and various minor criteria. We also show that our original recommendation of two systems involved with at least one with major criterion represents the minimal criteria because in probands not meeting these criteria, the yield of mutation identification drastically falls. This recommendation should help clinicians and biologists in identifying probands with a high probability of carrying a FBN1 gene mutation, and thus optimize biological resources. PMID- 19293844 TI - Use of advanced cluster analysis to characterize fish consumption patterns and methylmercury dietary exposures from fish and other sea foods among pregnant women. AB - On account of the interspecies variability in contamination and nutrient contents, consumers must balance the risks and benefits of fish consumption through their choice of species, meal size and frequency. The objectives of this study were to better characterize the risk of methylmercury (MeHg) exposure in a sample of 161 French pregnant women consuming sea food, including fish, molluscs and crustaceans, and to explore the use of unsupervised statistical learning as an advanced type of cluster analysis to identify patterns of fish consumption that could predict exposure to MeHg and the coverage of the Recommended Daily Allowance for n-3 polyunsaturated fatty acid (PUFA). The proportion of about 5% of pregnant women exposed at levels higher than the tolerable weekly intake for MeHg is similar to that observed among women of childbearing age in earlier French studies. At the same time, only about 50% of the women reached the recommended intake of 500 mg/day n-3 PUFA. Cluster analysis of the fish consumption showed that they could be grouped in five major clusters that are largely predictable of the intake of both MeHg and n-3 PUFA. This study shows that a global increase in seafood consumption could lead to MeHg exposure above the toxicological limits for pregnant women, thereby questioning the overall balance between this potential risk and potential beneficial effects of n-3 PUFA intakes. Only pregnant women consuming a high proportion of fatty fish meet the n 3 PUFA intake requirements without exceeding the toxicological limit for MeHg. The clusters identified suggest that different intervention strategies may be needed to address the dual purpose of ensuring high PUFA intakes at acceptable MeHg exposures. PMID- 19293845 TI - Perchlorate exposure from infant formula and comparisons with the perchlorate reference dose. AB - Perchlorate exposure may be higher in infants compared with older persons, due to diet (infant formula) and body weight versus intake considerations. Our primary objective was to quantitatively assess perchlorate concentrations in commercially available powdered infant formulas (PIFs). Secondary objectives were: (1) to estimate exposure in infants under different dosing scenarios and compare them with the perchlorate reference dose (RfD); (2) estimate the perchlorate concentration in water used for preparing PIFs that would result in a dose exceeding the RfD; and (3) estimate iodine intakes from PIFs. We quantified perchlorate levels in three samples (different lot numbers) of reconstituted PIF (using perchlorate-free water) from commercial brands of PIF in each of the following categories: bovine milk-based with lactose, soy-based, bovine milk based but lactose-free, and elemental (typically consisting of synthetic amino acids). Exposure modeling was conducted to determine whether the RfD might be exceeded in 48 dosing scenarios that were dependent on age, centile energy intake per unit of body weight, body weight percentile, and PIF perchlorate concentration. We obtained three different samples in each of the five brands of bovine- and soy-based PIF, three different samples in each of the three brands of lactose-free PIF, and three different samples in two brands of elemental PIF. The results were as follows: bovine milk-based with lactose (1.72 microg/l, range: 0.68-5.05); soy-based (0.21 microg/l, range: 0.10-0.44); lactose-free (0.27 microg/l, range: 0.03-0.93); and elemental (0.18 microg/l, range: 0.08-0.4). Bovine milk-based PIFs with lactose had a significantly higher concentration of perchlorate (P<0.05) compared with all. Perchlorate was a contaminant of all commercially available PIFs tested. Bovine milk-based PIFs with lactose had a significantly higher perchlorate concentration perchlorate than soy, lactose free, and elemental PIFs. The perchlorate RfD may be exceeded when certain bovine milk-based PIFs are ingested and/or when PIFs are reconstituted with perchlorate contaminated water. PMID- 19293846 TI - An overview of measurement method tools available to communities for conducting exposure and cumulative risk assessments. AB - Community-based programs for assessing and mitigating environmental risks represent a challenge to participants because each brings a different level of understanding of the issues affecting the community. These programs often require the collaboration of several community sectors, such as community leaders, local governments and researchers. Once the primary concerns, community vulnerabilities and assets are identified, participants plan on how to address immediate actions, rank known risks, collect information to support decision making, set priorities and determine an evaluation process to assess the success of the actions taken. The evaluation process allows the community to develop new action plans based on the results obtained from earlier actions. Tracking the success of the community actions may be as simple as a visual/tangible result (e.g., cleaning a park) or as complex as the collection of specific measurements to track the reduction of toxic pollutants or to determine the presence of a specific contaminant. Recognizing that communities may need to perform measurements to meet their goals, this paper provides an overview of the available measurement methods for several chemicals and biologicals in relevant environmental samples to a community setting. The measurement methods are organized into several categories according to their level of complexity, estimated cost and sources. Community project technical advisors are encouraged to examine the objective(s) of the community to be addressed by a measurement collection effort and the level of confidence that needed for the data to make appropriate decisions. The tables provide a starting point for determining which measurement method may be appropriate for specific community needs. PMID- 19293847 TI - Dissipative ring solitons with vorticity. AB - We study dissipative ring solitons with vorticity in the frame of the (2+1) dimensional cubic-quintic complex Ginzburg-Landau equation. In dissipative media, radially symmetric ring structures with any vorticity m can be stable in a finite range of parameters. Beyond the region of stability, the solitons lose the radial symmetry but may remain stable, keeping the same value of the topological charge. We have found bifurcations into solitons with n-fold bending symmetry, with n independent on m. Solitons without circular symmetry can also display (m + 1) fold modulation behaviour. A sequence of bifurcations can transform the ring soliton into a pulsating or chaotic state which keeps the same value of the topological charge as the original ring. PMID- 19293848 TI - Photonic logic by linear unidirectional interference. AB - A novel concept of unidirectional optical interference is presented, inspired by a non-Hermitian formulation of quantum mechanics via Maxwell-Schrodinger equation analogy. This model is employed for the design of photonic devices controlling strong fields by weak ones, suitable for large-scale optical logic. Implementations of very low-power photonic logic based on linear unidirectional optical gratings are presented. A linear photonic inverter is demonstrated and its performance is optimized by numerical calculations. The simulations are performed by full-vectorial beam propagation methods showing high signal to noise ratios. PMID- 19293849 TI - Polarization modulation of stochastic electromagnetic beams on propagation through the turbulent atmosphere. AB - Analytic expression is derived for the cross-spectral density matrix of a stochastic electromagnetic beam truncated by a slit aperture and passing through the turbulent atmosphere. The new formula can be used in study of the modulation in the spectral degree of polarization of the electromagnetic Gaussian Schell model beam on propagation. We find that the spectral degree of polarization in the output plane can be directly controlled by the width of the slit aperture. The effect of polarization shaping is also illustrated by numerical examples. PMID- 19293850 TI - Periodic index-modulated plasma waveguide. AB - We demonstrate a wire-obstructed cluster flow technique for making periodically modulated plasma waveguides in hydrogen, nitrogen, and argon with sharp, stable voids as short as 50 microm with a period as small as 200 microm. These gaps persist as the plasma expands for the full lifetime of the waveguide. We demonstrate guided propagation at intensities up to 2 x 10(17) W/cm(2), limited by our laser energy currently available. This technique is useful for quasi-phase matching applications where index-modulated guides are superior to diameter modulated guides. PMID- 19293851 TI - Large bandwidth, highly efficient optical gratings through high index materials. AB - We analyze the diffraction characteristics of dielectric gratings that feature a high index grating layer, and devise, through rigorous numerical calculations, large bandwidth, highly efficient, high dispersion dielectric gratings in reflection, transmission, and immersed transmission geometry. A dielectric TIR grating is suggested, whose -1dB spectral bandwidth is doubled as compared to its fused silica equivalent. The short wavelength diffraction efficiency is additionally improved by allowing for slanted lamella. The grating surpasses a blazed gold grating over the full octave. An immersed transmission grating is devised, whose -1dB bandwidth is tripled as compared to its fused silica equivalent, and that surpasses an equivalent classical transmission grating over nearly the full octave. A transmission grating in the classical scattering geometry is suggested, that features a buried high index layer. This grating provides effectively 100% diffraction efficiency at its design wavelength, and surpasses an equivalent fused silica grating over the full octave. PMID- 19293852 TI - 55-fs pulse generation without wave-breaking from an all-fiber Erbium-doped ring laser. AB - We demonstrate the direct generation of 55 fs pulses from an all-fiber Erbium doped ring laser oscillator using the nonlinear polarization rotation mode locking. The average output power is 56.4 mW but limited by available pump power of 330 mW. The linear chirped pulse duration is 55 fs after recompression using standard single-mode fiber. The pulses show to resist optical wave breaking with a smooth spectrum without any side lobe and cw-breakthrough. This all-fiber laser exhibits relatively high transfer efficiency of 17% and the single pulse energy reaches 1.5 nJ. PMID- 19293853 TI - Simultaneous optical parametric oscillation and intracavity second-harmonic generation based on a hexagonally poled lithium tantalate. AB - Simultaneous optical parametric oscillation and intracavity second-harmonic generation based on a hexagonally poled lithium tantalate is reported. Both the optical parametric oscillation and the cascaded noncollinear second-order harmonic generation processes reach a high efficiency. A variety of possible self doubling optical parametric oscillation processes indicate this hexagonally poled lithium tantalate has potential applications as a compact multi-wavelength light source. PMID- 19293854 TI - Multimode waveguide-cavity sensor based on fringe visibility detection. AB - Fringe visibility detection of the interaction of two bus spatial eigenmodes with a resonant cavity is investigated for the purpose of achieving a sensor platform with high sensitivity. The power distribution between the bus waveguide eigenmodes is modulated by the interaction with the cavity and is detected via fringe visibility lineshapes produced by twin-fiber interferometry. A test device is fabricated in a polymer-silica material system by a photolithographic process and is characterized by measuring the fringe visibility change as a function of analyte refractive index. Fringe visibility modulation from a straight two-mode waveguide coupled to a single mode ring resonator exposed to an aqueous glucose solution demonstrates a visibility change of 1.57 per weight percent, compared to a transmission change of 0.19 per weight percent for a single mode waveguide critically coupled to a ring with similar intrinsic quality factor. The demonstrated change in fringe visibility is 8.2 times larger. PMID- 19293855 TI - Theoretical investigation of polarization-insensitive data format conversion of RZ-OOK to RZ-BPSK in a nonlinear birefringent fiber. AB - Polarization-insensitive conversion of return-to-zero (RZ) ON-OFF keying (RZ-OOK) to RZ binary phase-shift keying (RZ-BPSK) has been achieved by cross-phase modulation (XPM) in a nonlinear birefringent fiber. This work presents a theoretical analysis of the dependence of format conversion on pump-probe detuning, and the pump state-of-polarization (SOP) that can fluctuate unpredictably in a realistic system. An investigation of the impact of pump polarization fluctuation on receiver sensitivity and receiver optimal threshold for the converted RZ-BPSK probe is also carried out. It was found that although the desired XPM-induced pi phase shift can be achieved by launching both the RZ OOK pump and the probe along the same birefringent axis of the fiber, the phase shift degrades to pi/3 if the SOP of the RZ-OOK pump unpredictably switches to the other axis of the fiber, resulting in a large receiver sensitivity penalty fluctuation of 14 dB. By contrast, launching the probe at 45 degrees relative to the birefringent axes can reduce the polarization-dependent receiver sensitivity penalty fluctuation to about 2 dB as the SOP of the RZ-OOK pump is swept over the Poincare sphere. These conclusions are in good agreement with recently. PMID- 19293856 TI - A negative-positive tunable liquid-crystal microlens array by printing. AB - A tunable microlens array by printing was demonstrated. An UV-curable adhesive, NOA65, was printed and cured to form a lens array profile on an ITO glass. Then this microlens array ITO glass was assembled with a normal ITO glass to form a cell, which was later filled with a liquid crystal. The focal length of the lens array can be tuned by an electric field, which changes the index difference between liquid crystals and NOA65 due to the reorientation of the LC molecules. In our experiment, the focal length varied from -2.29 cm to 3.12 cm when the applied voltage was increased from 0 V to 13.26 V. PMID- 19293857 TI - High-speed optical sampling using a silicon-chip temporal magnifier. AB - We demonstrate a single-shot technique for optical sampling based on temporal magnification using a silicon-chip time lens. We demonstrate the largest reported temporal magnification factor yet achieved (>500) and apply this technique to perform 1.3 TS/s single-shot sampling of ultrafast waveforms and to 80-Gb/s performance monitoring. This scheme offers the potential of developing a device that can transform GHz oscilloscopes into instruments capable of measuring signals with THz bandwidths. PMID- 19293858 TI - Compact efficient Q-switched eye-safe laser at 1525 nm with a double-end diffusion-bonded Nd:YVO4 crystal as a self-Raman medium. AB - We report on an efficient Q-switched eye-safe laser at 1525 nm with a double-end diffusion-bonded Nd:YVO(4) crystal as a self-Raman gain medium. A diffusion bonded crystal not only reduces the thermal effects but also increase the interaction length for the stimulated Raman scattering. With an input pump power of 17.2 W, average power of 2.23 W at the first- Stokes wavelength of 1525 nm is generated at a pulse repetition rate of 40 kHz, corresponding to a conversion efficiency of 13%. PMID- 19293859 TI - Tunable dispersion slope compensator using two uniform fiber Bragg gratings mounted on S-shape plate. AB - We propose and experimentally demonstrate a novel method for tunable dispersion slope compensation. We use two uniform fiber Bragg gratings (FBGs), a spatially designed S-bending stage and 4-port circulator. Two FBGs are mounted on each surface of a metal plate along the calculated quadratic curve. The dispersion slope (DS) can be tuned by adjusting a nonlinear strain along two uniform FBGs without changing second order dispersion as well as the central wavelength. In the experiment, a DS tuning range from -13.9 to -54.8 ps/nm(2) is achieved with the bandwidth of larger than 2.0 nm. PMID- 19293860 TI - Selectively enhanced band gap emission in ZnO/Ag2O nanocomposites. AB - A new composite consisting of ZnO nanorods decorated with Ag(2)O nanoparticles has been synthesized and characterized. It is found that the band gap emission of ZnO nanorods can be greatly enhanced by about 10 times, while the defect emission can be suppressed to the detection limit, simultaneously. The ratio between the band gap and defect emission reaches to an enhanced factor of about 600 times. The underlying mechanism is attributed to the combined effects of surface modification, band alignment, as well as charge transfer. Our approach provided here can be extended to many other semiconductors for creating nanocomposites with novel optical properties. PMID- 19293861 TI - Light wheel confined in a purely dielectric composite waveguide. AB - A properly designed composite waveguide consisting of a one-dimensional photonic crystal waveguide and a conventional dielectric waveguide is proposed for the realization of a localized "light wheel". Light confinedly rotating between the two waveguides is numerically demonstrated and explained physically in detail. A delocalized "light wheel" is found at the band gap edge caused by contra directional coupling between the two waveguides. Because of this delocalized "light wheel" , the composite waveguide can be used to trap light as a cavity, and a quality factor of 9 x 10(3) is achieved as an example. The present structure is completely dielectric and thus easy to realize with a low loss. PMID- 19293862 TI - Quantum cascade lasers and the Kruse model in free space optical communication. AB - Mid-infrared (MIR) free space optical communication has seen renewed interest in recent years due to advances in quantum cascade lasers. We present data from a multi-wavelength test-bed operated in the New York metropolitan area under realistic weather conditions. We show that a mid-infrared source (8.1 microm) provides enhanced link stability with 2x to 3x greater transmission over near infrared wavelengths (1.3 microm & 1.5 microm) during fog formation and up to 10x after a short scavenging rain event where fog developed and visibility reduced to approximately 1 km. We attribute the improvement to less Mie scattering at longer wavelengths. We confirm that this result is generally consistent with the empirical benchmark Kruse model at visibilities above 2.5 km, but towards the 1 km eye-seeing limit we measured the equivalent MIR visibility to be > 10 km. PMID- 19293863 TI - Magnetically tunable left handed metamaterials by liquid crystal orientation. AB - The tunability of an omega-type left handed metamaterial was demonstrated at microwave frequencies via the magnetic control of liquid crystal (LC) orientation. From the experimental and simulation results, it is shown that the left handed pass-band can be tuned by 220 MHz by changing the orientation of LC molecules by 90 degrees. A maximum index variation of 0.25 was obtained in the negative index regime with a measured LC birefringence of 0.05 in the 10 - 12 GHz frequency band. PMID- 19293864 TI - Optical sensitivity analysis of deformed mirrors for microcantilever array IR imaging. AB - Optical sensitivity is a major issue to improve the sensor responsivity and the spatial resolution of uncooled optomechanical focal plane arrays (FPA). The optical sensitivity is closely related to the mirror length and the undesired mirror deformation induced from the imbalanced residual stresses in different layers. In this paper, the influences of mirror length and deformation on the optical sensitivity are discussed by Fourier Optics. Theoretical analysis and experiments demonstrate that the optical sensitivity is seriously degraded by undesired mirror deformation, and that there exists an optimal mirror length which makes the optical sensitivity achieve its maximum under a certain mirror deformation. Based on the results, an optimized mirror configuration is presented to increase the optical sensitivity of substrate-free bi-material microcantilever array (SFBMA). PMID- 19293865 TI - Broadband laser tunability of Nd3+ ions in 0.8CaSiO3-0.2Ca3(PO4)2 eutectic glass. AB - Site-selective spectroscopy and stimulated emission experiments performed in the (4)F(3/2)-->(4)I(11/2) laser transition of Nd(3+)-doped 0.8CaSiO(3-) 0.2Ca(3)(PO(4))(2) eutectic glass are presented. The spectral features of the excitation spectra and those of spontaneous and stimulated emissions reveal the existence of a very complex crystal field site distribution for Nd(3+) ions. As a consequence, the stimulated emission of Nd(3+) in this glass shows a tunability of about 10 nm as a function of excitation wavelength. PMID- 19293866 TI - Suppression of ripples on ablated Ni surface via a polarization grating. AB - Gratings were recorded on the surface of nickel by ablation without formation of ripples using an interference of two p-polarized femtosecond laser beams at a pi/4 angle of incidence. The mechanism of ripples' suppression is explained by formation of a polarization grating and by ablation at the locations where the polarization is normal to the Ni surface. The aspect ratio of the ablated grooves was approximately 3 with the period approximately 570 nm at the central wavelength of irradiation of 800 nm. This method is applicable for laser structuring of different materials and a recorded grating structure can be scaled with the irradiation wavelength. PMID- 19293867 TI - Absolute efficiency estimation of photon-number-resolving detectors using twin beams. AB - A nonclassical light source is used to demonstrate experimentally the absolute efficiency calibration of a photon-number-resolving detector. The photon-pair detector calibration method developed by Klyshko for single-photon detectors is generalized to take advantage of the higher dynamic range and additional information provided by photon-number-resolving detectors. This enables the use of brighter twin-beam sources including amplified pulse pumped sources, which increases the relevant signal and provides measurement redundancy, making the calibration more robust. PMID- 19293868 TI - Low threshold monocrystalline Nd:(Gd, Lu)2O3 channel waveguide laser. AB - We report the first waveguide laser based on a rare-earth-doped sesquioxide. A 2 microm thick lattice matched Nd(0.5%):(Gd, Lu)(2)O(3) film with a nearly atomically flat surface has been epitaxially grown on a Y(2)O(3) substrate, using pulsed laser deposition. The film has been structured with reactive ion etching and a rib channel waveguide laser has been realized. Laser radiation at 1075 nm and 1079 nm has been observed under 820-nm pumping. The laser possesses a threshold power of about 0.8 mW and a preliminary slope efficiency of 0.5% versus incident pump power. A maximum output power of 1.8 mW has been obtained for 370 mW incident pump power. PMID- 19293869 TI - Pitch-variable blazed grating consisting of freestanding silicon beams. AB - Theoretical analysis is presented for a pitch-variable blazed grating which consists of freestanding silicon beams. The pitch-variable blazed grating is implemented by combining silicon-on-insulator (SOI) technology with microelectronicmechanical system (MEMS) technology. The whole device is fabricated on a 10 microm silicon device layer to guarantee sufficient stiffness. The 4-level blazed surface profile is realized by combining a two-mask process with fast atom beam etching. Electrostatic combdrive microactuators with double folded springs are proposed to stretch the freestanding grating beams. In association with reactive ion etching and vapor HF release, the freestanding grating beams and the microactuators are obtained, and a Cr/Au film is deposited onto the blazed grating surfaces by a protective mask process to improve the diffracted power. Mechanical response and diffraction efficiency of fabricated devices are characterized and the experimental results indicate that the fabricated 4- level blazed gratings extend both the tuning range and the diffraction efficiency of the 1st diffraction order of present MEMS diffraction gratings. PMID- 19293870 TI - Room temperature efficient actively Q-switched Ho:YAP laser. AB - We report on efficient actively Q-switched Ho:YAP laser double-pass pumped by a 1.91-microm laser. At room temperature, when the incident pump power was 20.9 W, a maximum average output power of 10.9 W at 2118 nm was obtained at the repetition rate of 10 kHz, and this corresponds to a conversion efficiency of 52.2% and a slope efficiency of 63.5%. Moreover, a maximum pulse energy of approximately 1.1 mJ and a minimum pulse width of 31 ns were achieved, with the peak power of 35.5 kW. PMID- 19293871 TI - Total optical transmission through a small hole in a metal waveguide screen. AB - We present the theory of total optical transmission through a small hole in metal waveguide screen. Unlike past works on extraordinary optical transmission using arrays, there is only a single hole; yet, the theory predicts total transmission for a perfect electric conductor (not normalized to the hole size) 100% transmission, regardless of how small the hole. This is very surprising considering the usual application of Bethe's theory to waveguide apertures. Comprehensive numerical simulations agree well with the theory and their modal analysis supports the proposed evanescent-mode mechanism for total transmission. These simulations are extended to show the influence of realistic material response (including loss) at microwave and visible-infrared frequencies. Due to the strong resonant field localization and transmission from only a thin metal screen with a single hole, many promising applications arise for this phenomenon including filtering, sensing, plasma generation, nonlinear optics, spectroscopy, heating, optical trapping, near-field microscopy and cavity quantum electrodynamics. PMID- 19293872 TI - Biased liquid crystal infiltrated photonic bandgap fiber. AB - A simulation scheme for the transmission spectrum of a photonic crystal fiber infiltrated with a nematic liquid crystal and subject to an external bias is presented. The alignment of the biased liquid crystal is simulated using the finite element method to solve the relevant system of coupled partial differential equations. From the liquid crystal alignment the full tensorial dielectric permittivity in the capillaries is derived. The transmission spectrum for the photonic crystal fiber is obtained by solving the generalized eigenvalue problem deriving from Maxwell's equations using a vector element based finite element method. We demonstrate results for a splay aligned liquid crystal infiltrated into the capillaries of a four-ring photonic crystal fiber and compare them to corresponding experiments. PMID- 19293873 TI - Dual-conjugate adaptive optics for wide-field high-resolution retinal imaging. AB - We present analysis and preliminary laboratory testing of a real-time dual conjugate adaptive optics (DCAO) instrument for ophthalmology that will enable wide-field high resolution imaging of the retina in vivo. The setup comprises five retinal guide stars (GS) and two deformable mirrors (DM), one conjugate to the pupil and one conjugate to a plane close to the retina. The DCAO instrument has a closed-loop wavefront sensing wavelength of 834 nm and an imaging wavelength of 575 nm. It incorporates an array of collimator lenses to spatially filter the light from all guide stars using one adjustable iris, and images the Hartmann patterns of multiple reference sources on a single detector. Zemax simulations were performed at 834 nm and 575 nm with the Navarro 99 and the Liou- Brennan eye models. Two correction alternatives were evaluated; conventional single conjugate AO (SCAO, using one GS and a pupil DM) and DCAO (using multiple GS and two DM). Zemax simulations at 575 nm based on the Navarro 99 eye model show that the diameter of the corrected field of view for diffraction-limited imaging (Strehl >or= 0.8) increases from 1.5 deg with SCAO to 6.5 deg using DCAO. The increase for the less stringent condition of a wavefront error of 1 rad or less (Strehl >or= 0.37) is from 3 deg with SCAO to approximately 7.4 deg using DCAO. Corresponding results for the Liou-Brennan eye model are 3.1 deg (SCAO) and 8.2 deg (DCAO) for Strehl >or= 0.8, and 4.8 deg (SCAO) and 9.6 deg (DCAO) for Strehl >or= 0.37. Potential gain in corrected field of view with DCAO is confirmed both by laboratory experiments on a model eye and by preliminary in vivo imaging of a human eye. PMID- 19293874 TI - Hybrid differential interrogation method for sensitive surface plasmon resonance measurement enabled by electro-optically tunable SPR sensors. AB - A novel detection method enabled by electro-optically tunable waveguide-coupled surface plasmon resonance sensors is demonstrated. Both the WCSPR response of sensor and the interrogation light are varied simultaneously in this hybrid scheme. Modulation and demodulation of the sensor's response are achieved by applying a high-frequency AC electrical signal and electrically filtering the detected signal. Scanning the incident angle at a lower speed yields an angular dependent response. Theoretical analyses and experimental results show that the angular-dependent signal is closely related to the derivative of the SPR reflectivity with a sharp, linear jump near the minimum of the SPR peak. Thus, simple linear-fitting and zero-finding algorithms can be used to locate the SPR angle, and sophisticated data processing algorithms and electronic hardware can be avoided. PMID- 19293875 TI - BER performance analysis of radio over free-space optical systems considering laser phase noise under Gamma-Gamma turbulence channels. AB - This paper analytically investigates a bit error rate (BER) performance of radio over free space optical (FSO) systems considering laser phase noise under Gamma Gamma turbulence channels. An external modulation using a dual drive Mach-Zehnder modulator (DD-MZM) and a phase shifter is employed because a DD-MZM is robust against a laser chirp and provides high spectral efficiency. We derive a closed form average BER as a function of different turbulence strengths and laser diode (LD) linewidth, and investigate its analytical behavior under practical scenario. As a result, for a given average SNR with normalized perturbation, it is shown that the difference of average BER corresponding to two LDs (with linewidth of 624 MHz and 10 MHz) under weak turbulence is almost 3 times larger than that under strong turbulence. PMID- 19293876 TI - Single mode fiber birefringence compensation in Sagnac and "plug & play" interferometric setups. AB - Single mode fiber (SMF) birefringence effects have been a limiting factor for a variety of Sagnac applications over longer distanceSMF links. In this report, we present a new concept of the SMF birefringence compensation in a Sagnac interferometric setup, based on a novel polarization control system. For the destructive interference, our control system guarantees a perfect compensation of both the SMF birefringence and imperfect propagation times matching of the setup's components. For the stabilization of the constructive interference, we have applied a fiber stretcher and a simple proportional-integral-derivative (PID) controller. The enclosed experimental data of the setup's visibility confirm validity of our polarization control system. We have also showed that the SMF birefringence model used in a "plug & play" interferometric setup [19], widely cited in the papers on quantum key distribution [11, 12, 13], cannot be applied in SMF Sagnac interferometric setup. However, the SMF birefringence model based on the Kapron equivalence well describes SMF Sagnac. PMID- 19293877 TI - White-light interference microscopy: minimization of spurious diffraction effects by geometric phase-shifting. AB - A common problem when profiling surfaces with steps or discontinuities using white-light (coherence-probe) interferometry is localized spikes (batwings) or spurious peaks due to diffraction effects. We show that errors due to these effects can be minimized by processing the irradiance data obtained with an achromatic phase-shifter operating on the geometric (Pancharatnam) phase to yield the values of the surface height. PMID- 19293878 TI - In-vivo demonstration of a high resolution optical fiber manometry catheter for diagnosis of gastrointestinal motility disorders. AB - Fiber optic catheters for the diagnosis of gastrointestinal motility disorders are demonstrated in-vitro and in-vivo. Single element catheters have been verified against existing solid state catheters and a multi-element catheter has been demonstrated for localized and full esophageal monitoring. The multi-element catheter consists of a series of closely spaced pressure sensors that pick up the peristaltic wave traveling along the gastrointestinal (GI) tract. The sensors are spaced on a 10 mm pitch allowing a full interpolated image of intraluminal pressure to be generated. Details are given of in-vivo trials of a 32-element catheter in the human oesophagus and the suitability of similar catheters for clinical evaluation in other regions of the human digestive tract is discussed. The fiber optic catheter is significantly smaller and more flexible than similar commercially available devices making intubation easier and improving patient tolerance during diagnostic procedures. PMID- 19293879 TI - Revealing single emitter spectral dynamics from intensity correlations in an ensemble fluorescence spectrum. AB - We show that the single emitter linewidth underlying a broadened ensemble emission spectrum can be extracted from correlations among the stochastic intensity fluctuations in the ensemble spectrum. Spectral correlations can be observed at high temporal and spectral resolutions with a cross-correlated pair of avalanche photodiodes placed at the outputs of a scanning Michelson interferometer. As illustrated with simulations in conjunction with Fluorescence Correlation Spectroscopy, our approach overcomes ensemble and temporal inhomogeneous broadening to provide single emitter linewidths, even for emitters under weak, continuous, broadband excitation. PMID- 19293880 TI - Mitigation of signal fading in radio over fiber transmission using fiber nonlinearity. AB - Radio over fiber systems in a microwave/millimeter wave band experience a severe signal fading due to fiber dispersion. However, parametric amplification in transmission fibers, known as modulation instability, can compensate for the signal fading. In this paper, we experimentally demonstrate radio over fiber transmission of 111.689 Mbps BPSK signal with a carrier frequency of 10.804 GHz, and measure bit error rates varying the optical transmission power. For the fiber launched power of 0 dBm, we observe a power penalty of 5.8 dB in the transmission over a 25 km fiber, though by increasing the fiber launched power up to +10 dBm, we successfully reduce the power penalty by 1.1 dB. PMID- 19293881 TI - Square nanosecond mode-locked Er-fiber laser synchronized to a picosecond Yb fiber laser. AB - A mode-locked Er-doped fiber laser was triggered to synchronize with a separate ultrashort picosecond Yb-doped fiber laser by cross phase modulation. Square nanosecond pulses were generated in the long-cavity Er-fiber laser by the peak intensity clamp effect while the synchronization maintained. At the maximum pump power of 450 mW, a synchronous laser pulse duration of 5.5 ns has been achieved. This synchronous nanosecond and picosecond system has shown a large length mismatch tolerance of 2.6 mm and can work stably for days. PMID- 19293882 TI - CO2 laser writing of long-period fiber grating in photonic crystal fiber under tension. AB - We demonstrate that the efficiency of CO(2) laser writing of long-period fiber gratings in a solid-core photonic crystal fiber (PCF) can be enhanced greatly by applying tension to the fiber during the writing process through the mechanism of frozen-in viscoelasticity. Using this mechanism, we are able to write strong gratings in PCFs with a dosage of CO(2) laser radiation low enough not to cause any significant fiber structure deformation. PMID- 19293883 TI - Alternative phase-diverse phase retrieval algorithm based on Levenberg-Marquardt nonlinear optimization. AB - A modified Levenberg-Marquardt (MLM) algorithm is proposed to substitute for modified G-S (MGS) algorithm in some situations of phase-diverse phase retrieval wavefront sensing (WFS), such as the obstructed pupil, in which the second derivative information is specifically employed to eliminate the local minimum stagnation. Experiments have been performed to validate MLM algorithm in WFS accuracy (less than lambda/30 RMS) referring to ZYGO interferometer results and in WFS repeatability (less than lambda/200 RMS), even the dynamic range is more than 7 lambda PV. Moreover, experiments have shown the MLM algorithm is superior to the MGS algorithm both in WFS accuracy and repeatability. PMID- 19293884 TI - Wide band gap materials as a new tuning strategy for dye doped cholesteric liquid crystals laser. AB - A new tuning strategy for mirror-less liquid crystals laser is presented. A three layer cell is prepared with two cholesteric layers sandwiching a layer containing an isotropic mixture of a photoluminescent dye. One of the chiral layers contains a wide band gap material while the second layer consists of a series of small band gap materials. Through the combination of these two layers, a set of mirrors that can selectively reflect different wavelengths is obtained. A different laser wavelength is emitted from different regions of the cell under the pumping beam irradiation. PMID- 19293885 TI - Stacked optical code label for multicasting in optical packet switching networks. AB - Stacked optical code (OC) label and its en/decoder based on fiber Bragg gratings (FBG) are proposed and experimentally demonstrated. This kind of label can carry several nodes' address information simultaneously in optical packet switching networks, so it can be employed in optical multicasting and simplify the node's structure a lot. The en/decoder is fabricated with high precision by our FBG techniques, and the experiment results show that the stacked OC label can support optical multicasting very well. PMID- 19293886 TI - Cantilever couplers for intra-chip coupling to silicon photonic integrated circuits. AB - An intra-chip coupling scheme from optical fibers to silicon strip waveguides is demonstrated. The couplers consist of silicon inverse width tapers embedded within silicon dioxide cantilevers that are deflected out-of-plane by residual stress. Deflection angles from 5 to 30 degrees are obtained and controlled by thermal annealing. Butt-coupling from tapered fibers or collimation-coupling from lensed fibers may be employed. The coupling scheme enables direct access to devices on the entire chip surface without dicing or cleaving the chip. Coupling efficiencies of 1.6 dB per connection for TE polarization and 2 dB per connection for TM polarization are achieved. The coupling efficiency shows little wavelength dependence, with less than 1.6 dB fluctuation over the wavelength range of 1500 nm to 1560 nm. PMID- 19293887 TI - Polarization-gated surface enhanced optical fields for ultrafast electron acceleration. AB - We show that the field polarization of the surface-plasmon-resonance enhanced optical field can be controlled to be linear with doubled intensity enhancement by using the polarization-gated excitation scheme with two counter-incident femtosecond laser pulses under the Kretschmann configuration, which is hence used for ultrafast electron acceleration to increase the maximum kinetic energy. The spatiotemporal evolution of the polarization-gated surface-enhanced optical field is studied by means of a simplified analytical model to describe the dynamical processes of electron acceleration, the kinetic energy and emission angular distributions of the accelerated electrons. PMID- 19293888 TI - Highly sensitive setup for tunable wavelength hyper-Rayleigh scattering with parallel detection and calibration data for various solvents. AB - A very sensitive experimental setup for accurate wavelength-dependent hyper Rayleigh scattering (HRS) measurements of the molecular first hyperpolarizability beta in the broad fundamental wavelength range of 600 to 1800 nm is presented. The setup makes use of a stable continuously tunable picosecond optical parametric amplifier with kilohertz repetition rate. To correct for multi-photon fluorescence, a small spectral range around the second harmonic wavelength is detected in parallel using a spectrograph coupled to an intensified charge coupled device. Reliable calibration against the pure solvent is possible over the full accessible spectral range. An extensive set of wavelength-dependent HRS calibration data for a wide range of solvents is presented, and very accurate measurements of the beta dispersion of the well-known nonlinear optical chromophore Disperse Red 1 are demonstrated. PMID- 19293889 TI - In-band crosstalk penalties in optical networks with narrow optical and electric filtering. AB - Optical cross-connects (OXC) introduce crosstalk and intersymbolic interference due to filtering in the optical signal. We show experimentally and by simulation that higher optical signal-to-noise ratio (OSNR) penalties are obtained for the combined effect of both impairments compared to the sum of the penalties taken independently. Furthermore, the Modified Chernoff Bound is applied for performance estimation, with good accuracy and fast computation time. By using the correct OSNR penalty, the maximum number of cascaded OXC is reduced by 17% relatively to when the penalties are considered separately, for a 2 dB maximum penalty. PMID- 19293890 TI - Characterizing isolated attosecond pulses from hollow-core waveguides using multi cycle driving pulses. AB - The generation of attosecond-duration light pulses using the high-order harmonic generation process is a rapidly evolving area of research. In this work, we combine experimental measurements with careful numerical analysis, to demonstrate that even relatively long-duration, 15 fs, carrier-envelope-phase (CEP) unstabilized near-infrared (NIR) pulses can generate isolated attosecond extreme ultraviolet (EUV) pulses by the dynamically-changing phase matching conditions in a hollow-core waveguide geometry. The measurements are made using the laser assisted photoelectric effect to cross-correlate the EUV pulse with the NIR pulse. A FROG CRAB analysis of the resulting traces (photoelectron signal versus photoelectron energy and EUV-NIR delay) is performed using a generalized projections (GP) algorithm, adapted for a wide-angle photoelectron detection geometry and non-CEP stabilized driving laser pulses. In addition, we performed direct FROG CRAB simulations under the same conditions. Such direct simulations allow more freedom to explore the effect of specific pulse parameters on FROG CRAB traces than is possible using the automated GP retrieval algorithm. Our analysis shows that an isolated pulse with duration of approximately 200 attoseconds can result from CEP unstabilized, high intensity approximately 15 fs multi-cycle driving pulses coupled into a hollow-core waveguide filled with low pressure Argon gas. These are significantly longer driving pulses than used in other experimental implementations of isolated attosecond pulses. PMID- 19293891 TI - Fourier transform spectroscopy using stressed liquid crystal. AB - A Stressed Liquid Crystal (SLC) is proposed for application as a single panel retardance element in a Fourier transform (FT) spectrometer. Volume alignment in SLCs increase the maximum retardance and subsequent FT resolution by creating greater path lengths. Here, the relationship between transmission and shear for thick SLC cells is characterized and the spectral resolution using the SLC phase modulators in a single and double pass FT spectrometer system is quantified. For a 100 microm thick SLC, the resolution of a single frequency peak was observed at 60 nm full width half maximum. PMID- 19293892 TI - Low-power all-optical switch based on the bend effect of a nm fiber taper driven by outgoing light. AB - We demonstrate a very simple all-optical switch (AOS) based on the bend effect of a nm fiber taper driven by outgoing light. The AOS operates at relative low control power compared with those of using nonlinear effect. In our configuration, we find that the turnoff and turnon time of the AOS are about 500 ms and 760 ms, respectively. The optimized extinction ratio of about 15 db under 32 mW control light at 980 nm is achieved. This AOS effect will be very useful for creation of different interesting functional devices. PMID- 19293893 TI - Broadband antireflective poly-Si nanosponge for thin film solar cells. AB - Antireflective nanosponges are fabricated on polycrystalline silicon (poly-Si) thin films using Ag-nanoparticles (NPs) assisted etching. Crystal orientations and grain sizes of the poly-Si thin films are investigated for the poly-Si nanosponge formation and the resultant optical properties. The Ag-NPs assisted etching preferentially etches the poly-Si thin films along crystal orientation of [110]. A 400 nm thick poly-Si nanosponge reduces effective optical reflection of the poly-Si thin film with substrate crystal orientation of (110) and averaged grain size of 250 nm from 26 % to 3 % at the wavelengths ranging from 400 nm to 1000 nm. Carrier lifetimes were found to be 41 and 36 mus for poly-Si thin film and RTO-passivated nanosponges, respectively. PMID- 19293894 TI - Coherent nonlinear emission from a single KTP nanoparticle with broadband femtosecond pulses. AB - We demonstrate that the intensity of the second harmonic (SH) generated in KTiOPO(4) nanoparticles excited with femtosecond laser pulses increases with decreasing duration of the infrared pump pulses. The SH intensity scales, approximately, as the inverse of the laser pulse duration ranging between 13 fs and 200 fs. The SH intensity enhancement requires careful compensation of the high-order spectral phase, being achieved with a genetic algorithm. Using ultrashort laser pulses improves the signal-to-noise ratio and will allow the detection of 10-nm size particles. Finally, we demonstrate that the spectrum of broadband (100 nm) pulses can be shaped to generate non-degenerate sum-frequency mixing. This opens up access to the polarization degrees of freedom of this second-order nonlinear process at the nanoscale. PMID- 19293895 TI - Evaporation kinetics of a non-spherical, levitated aerosol particle using optical resonance spectroscopy for precision sizing. AB - We describe how a time series of optical resonance spectra of an evaporating, non spherical, irregular aerosol particle levitated in an electrodynamic balance exhibits patterns which are related to its evaporation kinetics. Simulated spectra of an evaporating, model aerosol particle show comparable features. If these patterns are used to deduce the particle size change with time, the resulting vapor pressures and enthalpies of vaporization compare favorably with literature data for both crystalline ammonium nitrate and succinic acid particles. PMID- 19293896 TI - Nonclassical 2-photon interference with separate intrinsically narrowband fibre sources. AB - In this paper, we demonstrate a source of photon pairs based on four-wave-mixing in photonic crystal fibres. Careful engineering of the phase matching conditions in the fibres enables us to create photon pairs at 597 nm and 860 nm in an intrinsically factorable state showing no spectral correlations. This allows for heralding one photon in a pure state and hence renders narrow band filtering obsolete. The source is narrow band, bright and achieves an overall detection efficiency of up to 21% per photon. For the first time, a Hong-Ou-Mandel interference with unfiltered photons from separate fibre sources is presented. PMID- 19293897 TI - Estimation of space-borne lidar return from natural waters: a passive approach. AB - A method, based on the reciprocity principle of radiative transfer, for using routinely collected field measurements of apparent optical properties in a water body to estimate the total return (time integrated) to an airborne or space borne lidar is presented. It will allow prediction of lidar returns using the databases of apparent optical propertie assembled in support of ocean color remote sensing. PMID- 19293898 TI - Accurate detection and complete tracking of large populations of features in three dimensions. AB - Localization and tracking of colloidal particles in microscopy images generates the raw data necessary to understand both the dynamics and the mechanical properties of colloidal model systems. Yet, despite the obvious importance of analyzing particle movement in three dimensions (3D), accurate sub-pixel localization of the particles in 3D has received little attention so far. Tracking has been limited by the choice of whether to track all particles in a low-density system, or whether to neglect the most mobile fraction of particles in a dense system. Moreover, assertions are frequently made on the accuracies of methods for locating particles in colloid physics and in biology, and the field of particle locating and tracking can be well-served by quantitative comparison of relative performances. We show that by iterating sub-pixel localization in three dimensions, the centers of particles can be more accurately located in three-dimensions (3D) than with all previous methods by at least half an order of magnitude. In addition, we show that implementing a multi-pass deflation approach, greater fidelity can be achieved in reconstruction of trajectories, once particle positions are known. In general, all future work must defend the accuracy of the particle tracks to be considered reliable. Specifically, other researchers must use the methods presented here (or an alternative whose accuracy can be substantianted) in order for the entire investigation to be considered legitimate, if the basis of the physical argument (in colloids, biology, or any other application) depends on quantitative accuracy of particle positions. We compare our algorithms to other recent and related advances in location/tracking in colloids and in biology, and discuss the relative strengths and weaknesses of all the algorithms in various situations. We carry out performance tests directly comparing the accuracy of our and other 3D methods with simulated data for both location and tracking, and in providing relative performance data, we assess just how accurately software can locate particles. We discuss how our methods, now applied to colloids, could improve the location and tracking of features such as quantum dots in cells. PMID- 19293899 TI - The generalized Wolf shift for cyclostationary fields. AB - Correlation dependent, propagation-induced shifts in the generalized spectra of cyclostationary, random fields are predicted. This result generalizes the Wolf shift for stationary fields and is applicable to periodic trains of fast pulses such as might be generated in comb spectroscopy or other mode-locked pulsed systems. Examples illustrate these shifts for intrinsically stationary fields and the fields generated by a mode-locked laser. PMID- 19293900 TI - Electrically pumped ultraviolet random lasing from ZnO-based metal-insulator semiconductor devices: dependence on carrier transport. AB - The electrically pumped ultraviolet (UV) random lasing and carrier transport of ZnO-based metal-insulator-semiconductor (MIS) structures on Si substrates have been systematically investigated. With the increase of positive bias voltage on the gates of the MIS devices, the current-voltage (I-V) characteristics manifest a normal curved I-V region where the current increases with the bias, followed by a negative differential resistance (NDR) region. Moreover, the UV electroluminescence from the devices in the normal region is transformed from spontaneous emission into increasingly intensive random lasing; while, that in the NDR region is transformed from random lasing into very weak spontaneous emission. The reason for the effect of NDR on the random lasing from the devices has been tentatively explored. PMID- 19293901 TI - Analysis and fabrication of a prism film with roll-to-roll fabrication process. AB - A prism-based brightness enhancement film (BEF), has been investigated through computer-aided optical analysis and experiments in this study. The prism film was fabricated on a 100 microm thick PET film substrate using roll-to-roll (R2R) process with UV curable resins of different refractive indices. The results from both optical analysis and experiments showed that resins with higher refractive indices have better on-axis luminous gain. The prism structures fabricated on the substrate achieved a replication rate of 97.54% in this study. Compared with the 3M BEF, the prism films developed in this study are improved in on-axis luminous gains when the refractive index of the resin is higher than 1.53. PMID- 19293902 TI - Ultraviolet-visible conical emission by multiple laser filaments. AB - We characterized the angular distribution of the supercontinuum emission from multiple infrared laser filaments propagating in air over long distances, from the infrared (1080 nm) to ultraviolet (225 nm). These experimental data suggest that the X-Waves modeling or Cerenkov emission, rather than phase matching of four-wave mixing, could explain the conical emission. We also estimate the total light conversion efficiency from the original laser wavelength into the white light continuum. PMID- 19293903 TI - Physical limits to broadening compensation in a linear slow light system. AB - The dispersion experienced by a signal in a slow light system leads to a significant pulse broadening and sets a limit to the maximum delay actually achievable by the system. To overcome this limitation, a substantial research effort is currently being carried out, and successful strategies to reduce distortion in linear slow light systems have already been demonstrated. Recent theoretical and experimental works have even claimed the achievement of zero broadening of pulses in these systems. In this work we obtain some physical limits to broadening compensation in linear slow light systems based on simple Fourier analysis. We show that gain and dispersion broadening can never compensate in such a system. Additionally, it is simply proven that all the linear slow light systems that introduce a low-pass filtering of the signal (a reduction in the signal root-mean- square spectral width), will always cause pulse broadening. These demonstrations are done using a rigorous shape independent definition of pulse width (the root-mean-square temporal width) and arguments borrowed from time-frequency analysis. PMID- 19293904 TI - Subcarrier multiplexing tolerant dispersion transmission system employing optical broadband sources. AB - This paper presents a novel SCM optical transmission system for next-generation WDM-PONs combining broadband optical sources and a Mach-Zehnder interferometric structure. The approach leeds to transport RF signals up to 50 GHz being compatible with RoF systems since a second configuration has been proposed in order to overcome dispersion carrier suppression effect using DSB modulation. The theoretical analysis validates the potentiality of the system also considering the effects of the dispersion slope over the transmission window. PMID- 19293905 TI - Low loss etchless silicon photonic waveguides. AB - We demonstrate low loss silicon waveguides fabricated without any silicon etching. We define the waveguides by selective oxidation which produces ultra smooth sidewalls with width variations of 0.3 nm. The waveguides have a propagation loss of 0.3 dB/cm at 1.55 microm. The waveguide geometry enables low bending loss of approximately 0.007 dB/bend for a 90 degrees bend with a 50 microm bending radius. PMID- 19293906 TI - Cloaking radiation of moving electron beam and relativistic energy loss spectra. AB - The interaction of ideal and nonideal metamaterial cylindrical cloaks with an electron beam has been studied. The exact solution for energy loss suffered by a fast electron moving inside or outside a cylindrical cloak has been established within a fully relativistic approach. The effect of various imperfect parameters on the efficiency of the cloak has been discussed. It is shown that radiation can be shielded very well by such cloaks when an electron moves either inside or outside of them. The efficiency of nonideal cloaks and the effect of various nonideal parameters on cloak invisibility can be exhibited in the spectra of electron energy loss and radiation emission. This means that the property of a cylindrical cloak can be explored by means of scanning transmission electron microscopy. PMID- 19293907 TI - Opto-VLSI-based photonic true-time delay architecture for broadband adaptive nulling in phased array antennas. AB - This paper proposes a novel Opto-VLSI-based tunable true-time delay generation unit for adaptively steering the nulls of microwave phased array antennas. Arbitrary single or multiple true-time delays can simultaneously be synthesized for each antenna element by slicing an RF-modulated broadband optical source and routing specific sliced wavebands through an Opto-VLSI processor to a high dispersion fiber. Experimental results are presented, which demonstrate the principle of the true-time delay unit through the generation of 5 arbitrary true time delays of up to 2.5 ns each. PMID- 19293908 TI - A liquid-filled tunable double-focus microlens. AB - A novel microlens design with tunable double-focus is presented. It is fabricated by adding only one SU-8 photolithography step to the well-developed liquid-filled microlens fabrication process. The thickness of this layer determines the thickness difference between the central and peripheral region of the membrane, the deformation of which is used to define the surface profile of the microlens. The stepped thickness variation is finally manifested as the difference in deformation contour at two different regions of the membrane when subjected to uniform applied pressure, thereby causing two focal lengths to appear. Experimental and simulation results are presented, from which the tunability of the focal lengths of the double-focus microlens is demonstrated to be effective over a wide range through combining the structural design with pressure control. The successful demonstration of this unconventional microlens design concept will potentially extend t application of liquid-filled microlens technology. PMID- 19293909 TI - An optical power combiner/wavelength demultiplexing module for hybrid WDM FTTX. AB - An optical device scheme that serves simultaneously as a power combiner for upstream and wavelength demultiplexer for downstream signals is presented. The design concept is validated experimentally by an optical module based on off-the shelf discrete optical components. An integrated device based on planar lightwave circuit (PLC) is proposed and analyzed in which a multi-mode interference (MMI) device is utilized to separate the upstream 1310 nm signal from the downstream 155x nm signals. The dense WDM function is realized through an arrayed-waveguide grating (AWG). Design guidelines and optimization procedure for the device are discussed by way of examples. PMID- 19293910 TI - Collective coupling of atoms with cavity mode and free-space field. AB - We report an experimental study of the collective coupling of three-level atoms with a cavity mode and a free-space laser field. The measurement of the cavity transmission with a weak probe field coupled into the cavity mode reveals three spectral peaks: two sidebands and a central peak, which is produced by the coherent interaction of the free-space field with individual atoms and the collective interaction of the multiple atoms with the cavity mode. The experimental results agree with a simple calculation based on the classical light transmission through a cavity containing multiple atoms coherently driven by a free-space laser. PMID- 19293911 TI - Manipulation of operation states by polarization control in an erbium-doped fiber laser with a hybrid saturable absorber. AB - We propose an operation switchable ring-cavity erbium-doped fiber laser (EDFL) via intra-cavity polarization control. By using a semiconductor saturable absorber mirror in the EDFL cavity, stable Q-switching, Q-switched mode-locking, continuous-wave mode-locking, pulse splitting, and harmonic mode-locking pulses can be manipulated simply by detuning a polarization controller while keeping the pump power at the same level. All EDFL operation states can be obtained under the polarization angles detuning within 180 degrees. Continuous-wave mode-locking of EDFL with 800-fs pulsewidth repeated at 4 MHz has been obtained, for which the output pulse energy is 0.5 nJ and the peak power is 625 W. Interaction between solitons and the accompanied non-soliton component will lead to either pulse splitting or 5th-order harmonic mode-locking at repetition rate of 20 MHz. PMID- 19293912 TI - WDM coherent PDM-QPSK systems with and without inline optical dispersion compensation. AB - Using numerical simulations, we study and compare the performance of 42.8-Gb/s and 112-Gb/s intradyne coherent polarization-division- multiplexed quadrature phase-shift-keying (PDM-QPSK) systems in wavelength-division-multiplexed (WDM) transmission with inline dispersion compensation fiber (DCF) and that with fully electronic dispersion compensation. Two effects are considered in the studies. One is fiber nonlinearities and the other is the local oscillator (LO) phase noise to amplitude noise conversion induced by electronic dispersion compensation. Results of 1000-km transmission employing standard single-mode fiber (SSMF) show that, for non-return-to-zero (NRZ) PDM-QPSK, both the 42.8-Gb/s and 112-Gb/s WDM systems with DCF have less tolerance to fiber nonlinearities than those with electronic dispersion compensation due to nonlinear polarization scattering. However, by using time-interleaved return-to-zero (RZ) P -QPSK, which can significantly suppress nonlinear polarization scattering in a system with inline DCF, the 42.8-Gb/s system with DCF can achieve better performance than that with electronic dispersion compensation, and comparable performance can be obtained for the 112-Gb/s system with DCF and that with electronic dispersion compensation. We find that the LO phase noise to amplitude noise conversion can cause significant penalties in the 112-Gb/s system with only electronic dispersion compensation if distributed feedback lasers are used. PMID- 19293913 TI - Nanoscale optical field localization by resonantly focused plasmons. AB - We experimentally demonstrate use of plasmonic resonant phenomena combined with strong field localization to enhance efficiency of confining optical fields in a Si waveguide. Our approach utilizes a plasmonic resonant nano-focusing-antenna (RNFA), that simultaneously supports several focusing mechanisms in a single nanostructure, integrated with a lossless Si waveguide utilized with silicon-on insulator (SOI) technology, to achieve a sub-diffraction limited focusing with a nanoscale (deeply subwavelength) spot size. The metallic RNFA effectively converts an incoming propagating waveguide mode to a localized resonant plasmon mode in an ultrasmall volume in all 3 dimensions. The near-field optical measurements of the fabricated RNFA using heterodyne near-field scanning optical microscope (H-NSOM) validate the theoretical predictions showing strong optical field localization. PMID- 19293915 TI - Periodically poled Ti-diffused near-stoichiometric MgO:LiNbO3 waveguide nonlinear optic wavelength converter. AB - Periodically poled titanium-diffusion waveguide was fabricated in near stoichiometric MgO:LiNbO(3) wafers. The characteristics of the device were examined by pump-probe second harmonic generation (SHG). The device shows very high resistance to photorefractive damage at room temperature. The wavelength tuning of the converted difference frequency (DF) wave can be achieved from 1450 to 1542 nm by tuning pump wave and signal wave. The wavelength conversion efficiency was measured to be -7.3 dB with coupled pump and signal power are 150 mW and 50 mW, respectively. PMID- 19293916 TI - Anterior segment imaging with Spectral OCT system using a high-speed CMOS camera. AB - We describe a new ultrahigh speed Spectral OCT instrument making use of a CMOS camera and demonstrate high quality in vivo imaging of the anterior segment of the human eye. The high flexibility of the designed imaging system allows a wide range of imaging protocols. Two- and three-dimensional high quality OCT images of the cornea, the anterior chamber and the crystalline lens are presented. A high acquisition rate, up to 135,000 A-scans/second enables three-dimensional reconstruction of the anterior segment during lenticular accommodation, blinking and pupillary reaction to light stimulus. We demonstrate OCT tomographic real time imaging of the lens dynamics during accommodation and high quality OCT cross sectional images of the entire anterior segment of the eye from the cornea up to posterior part of the crystalline lens. PMID- 19293917 TI - Side-mode transmission diagnosis of a multichannel selectable injection-locked Fabry-Perot Laser Diode with anti-reflection coated front facet. AB - Theory and experiments on the side-mode-suppression-ratio (SMSR) enhancement and the linewidth reduction of a Fabry-Perot laser diode (FPLD) side-mode-injection locked by using another FPLD are demonstrated to realize its potential application as a DWDM transmitter source. The SMSR, the spectral linewidth and the linewidth enhancement factor are simulated to realize the limitation of the FPLD-FPLD link under side-mode injection-locking condition. A degradation of the linewidth enhancement factor from 1.5 to 2.1 is observed due to the slave FPLD injection-locked at principle- and side-mode conditions. Up to 22-channel selectability of the 2.5 Gbit/s directly modulated FPLD based transmitter under side-mode injection-locking is demonstrated with a SMSR >35 dB, a Q-factor 6.8 9.2, a locking range of 24 nm, a power penalty of -0.7 dB, and a BER of 10(-10) at -17 dBm. The side-mode injection-locked FPLD shows high-quality transmission performance and meet the demand for cost-effective and high-capability 2.5 Gbit/s WDM systems. PMID- 19293918 TI - Photothermally-induced disordered patterns of corneal collagen revealed by SHG imaging. AB - The loss of organization of the corneal collagen lattice induced by photothermal effects was analyzed by using second-harmonic generation (SHG) imaging. Porcine cornea samples were treated with low-power laser irradiation in order to get localized areas of tissue disorganization. The disorder induced within the irradiated area of corneal stroma was quantified by means of Discrete Fourier Transform, auto-correlation and entropy analyses of the SHG images. Polarization modulated SHG measurements allowed to probe the changes in the structural anisotropy of sub-micron hierarchical levels of the stromal collagen. Our results emphasize the great potential of the SHG imaging to detect subtle modifications in the collagen assembly. The proposed analytical methods may be used to track several genetic, pathologic, accidental or surgical-induced disorder states of biological tissues. PMID- 19293919 TI - Quantitative photoacoustic measurement of tissue optical absorption spectrum aided by an optical contrast agent. AB - In photoacoustic imaging, the intensity of photoacoustic signal induced by optical absorption in biological tissue is proportional to light energy deposition, which is the product of the absorption coefficient and the local light fluence. Because tissue optical properties are highly dependent on the wavelength, the spectrum of the local light fluence at a target tissue beneath the sample surface is different than the spectrum of the incident light fluence. Therefore, quantifying the tissue optical absorption spectrum by using a photoacoustic technique is not feasible without the knowledge of the local light fluence. In this work, a highly accurate photoacoustic measurement of the subsurface tissue optical absorption spectrum has been achieved for the first time by introducing an extrinsic optical contrast agent with known optical properties. From the photoacoustic measurements with and without the contrast agent, a quantified measurement of the chromophore absorption spectrum can be realized in a strongly scattering medium. Experiments on micro-flow vessels containing fresh canine blood buried in phantoms and chicken breast tissues were carried out in a wavelength range from 680 nm to 950 nm. Spectroscopic photoacoustic measurements of both oxygenated and deoxygenated blood specimens presented an improved match with the references when employing this technique. PMID- 19293920 TI - Single-frequency synthesis at telecommunication wavelengths. AB - We report an optical single-frequency synthesizer at the 1.55 microm telecommunications band. Output from a continuous-wave external cavity diode laser is frequency doubled and phase locked to a predetermined component of a Ti:S laser frequency comb. The synthesizer is capable of generating a single user specified frequency from an atomic time base within the 192-196 THz gain bandwidth of an erbium-doped fiber amplifier. By tuning the repetition rate of the femtosecond laser the synthesized optical frequency can be swept with sub kilohertz step size. Frequency sweeps of several GHz are realized by automatically re-locking the diode laser to adjacent comb components during frequency sweep. We demonstrate the operation of the device by presenting results of Doppler-free spectroscopy on acetylene using synthesized frequencies. PMID- 19293921 TI - Diffusion stabilizes cavity solitons in bidirectional lasers. AB - We study the influence of field diffusion on the spatial localized structures (cavity solitons) in bidirectional lasers. We find threefold positive role of the diffusion: 1) it allows for the existence of solitons in cavities with equal losses for the two fields; 2) it increases the stability range of the individual (isolated) solitons; and 3) it reduces the long-range interaction between cavity solitons, allowing for the independent manipulation (writing and erasing) of individual structures. PMID- 19293922 TI - Polarization-independent self-collimation based on pill-void photonic crystals with square symmetry. AB - We investigate discrepancy and similarity in dispersion relations between transverse-electric (TE) and transverse-magnetic (TM) polarizations in rectangular, square and triangular two-dimensional photonic crystals. It is found that the square lattice is the most appropriate candidate to realize polarization independent, i.e. absolute self-collimation (ASC) in the first photonic band since it possesses not only a relatively broad angular range for self-collimation but also a small difference in dispersion relations between TE and TM modes. By tailoring the shape of air voids in the square-lattice-based structure, the electric-field vector can be rotated to reduce the discrepancy between TE and TM modes whereby the frequency bandwidth of ASC can be enlarged to approximately 4.8%. The ASC phenomenon is demonstrated by numerical experiments based on a finite-difference time-domain (FDTD) technique with negligible propagation losses. PMID- 19293923 TI - Design of all-solid leakage channel fibers with large mode area and low bending loss. AB - We investigate a novel design for all-solid large mode area (LMA) leakage channel fibers (LCFs) for high-power Yb-doped fiber lasers and amplifiers, based on a single down-doped-silica rod ring surrounding a seven-cell pure-silica core, aiming for effectively single-mode behavior and low bending loss characteristics. Through detailed numerical simulations based on the finite element method (FEM), we find that the proposed all-solid LMA-LCFs, having a seven-cell core and two different sizes of down-doped rods, can achieve sufficient differential mode loss and much lower bending loss, as compared with a previously-reported LCF with a one-cell core and six large down-doped-silica rods. PMID- 19293924 TI - Media reporting of health interventions: signs of improvement, but major problems persist. AB - BACKGROUND: Studies have persistently shown deficiencies in medical reporting by the mainstream media. We have been monitoring the accuracy and comprehensiveness of medical news reporting in Australia since mid 2004. This analysis of more than 1200 stories in the Australian media compares different types of media outlets and examines reporting trends over time. METHODS AND FINDINGS: Between March 2004 and June 2008 1230 news stories were rated on a national medical news monitoring web site, Media Doctor Australia. These covered a variety of health interventions ranging from drugs, diagnostic tests and surgery to dietary and complementary therapies. Each story was independently assessed by two reviewers using ten criteria. Scores were expressed as percentages of total assessable items deemed satisfactory according to a coding guide. Analysis of variance was used to compare mean scores and Fishers exact test to compare proportions. Trends over time were analysed using un-weighted linear regression analysis. Broadsheet newspapers had the highest average satisfactory scores: 58% (95% CI 56-60%), compared with tabloid newspapers and online news outlets, 48% (95% CI 44-52) and 48% (95% CI 46-50) respectively. The lowest scores were assigned to stories broadcast by human interest/current affairs television programmes (average score 33% (95% CI 28-38)). While there was a non- significant increase in average scores for all outlets, a significant improvement was seen in the online news media: a rise of 5.1% (95%CI 1.32, 8.97; P 0.009). Statistically significant improvements were seen in coverage of the potential harms of interventions, the availability of treatment or diagnostic options, and accurate quantification of benefits. CONCLUSION: Although the overall quality of medical reporting in the general media remains poor, this study showed modest improvements in some areas. However, the most striking finding was the continuing very poor coverage of health news by commercial current affairs television programs. PMID- 19293926 TI - Leveraging rural energy investment for parasitic disease control: schistosome ova inactivation and energy co-benefits of anaerobic digesters in rural China. AB - BACKGROUND: Cooking and heating remain the most energy intensive activities among the world's poor, and thus improved access to clean energies for these tasks has been highlighted as a key requirement of attaining the major objectives of the UN Millennium Development Goals. A move towards clean energy technologies such as biogas systems (which produce methane from human and animal waste) has the potential to provide immediate benefits for the control of neglected tropical diseases. Here, an assessment of the parasitic disease and energy benefits of biogas systems in Sichuan Province, China, is presented, highlighting how the public health sector can leverage the proliferation of rural energy projects for infectious disease control. METHODOLOGY/FINDINGS: First, the effectiveness of biogas systems at inactivating and removing ova of the human parasite Schistosoma japonicum is experimentally evaluated. Second, the impact of biogas infrastructure on energy use and environmental quality as reported by surveyed village populations is assessed, as is the community acceptance of the technology. No viable eggs were recovered in the effluent collected weekly from biogas systems for two months following seeding with infected stool. Less than 1% of ova were recovered viable from a series of nylon bags seeded with ova, a 2-log removal attributable to biochemical inactivation. More than 90% of Ascaris lumbricoides ova (used as a proxy for S. japonicum ova) counted at the influent of two biogas systems were removed in the systems when adjusted for system residence time, an approximate 1-log removal attributable to sedimentation. Combined, these inactivation/removal processes underscore the promise of biogas infrastructure for reducing parasite contamination resulting from nightsoil use. When interviewed an average of 4 years after construction, villagers attributed large changes in fuel usage to the installation of biogas systems. Household coal usage decreased by 68%, wood by 74%, and crop waste by 6%. With reported energy savings valued at roughly 600 CNY per year, 2-3 years were required to recoup the capital costs of biogas systems. In villages without subsidies, no new biogas systems were implemented. CONCLUSIONS: Sustainable strategies that integrate rural energy needs and sanitation offer tremendous promise for long-term control of parasitic diseases, while simultaneously reducing energy costs and improving quality of life. Government policies can enhance the financial viability of such strategies by introducing fiscal incentives for joint sanitation/sustainable energy projects, along with their associated public outreach and education programs. PMID- 19293925 TI - Placebo response of non-pharmacological and pharmacological trials in major depression: a systematic review and meta-analysis. AB - BACKGROUND: Although meta-analyses have shown that placebo responses are large in Major Depressive Disorder (MDD) trials; the placebo response of devices such as repetitive transcranial magnetic stimulation (rTMS) has not been systematically assessed. We proposed to assess placebo responses in two categories of MDD trials: pharmacological (antidepressant drugs) and non-pharmacological (device- rTMS) trials. METHODOLOGY/PRINCIPAL FINDINGS: We performed a systematic review and meta-analysis of the literature from April 2002 to April 2008, searching MEDLINE, Cochrane, Scielo and CRISP electronic databases and reference lists from retrieved studies and conference abstracts. We used the keywords placebo and depression and escitalopram for pharmacological studies; and transcranial magnetic stimulation and depression and sham for non-pharmacological studies. All randomized, double-blinded, placebo-controlled, parallel articles on major depressive disorder were included. Forty-one studies met our inclusion criteria - 29 in the rTMS arm and 12 in the escitalopram arm. We extracted the mean and standard values of depression scores in the placebo group of each study. Then, we calculated the pooled effect size for escitalopram and rTMS arm separately, using Cohen's d as the measure of effect size. We found that placebo response are large for both escitalopram (Cohen's d - random-effects model - 1.48; 95%C.I. 1.26 to 1.6) and rTMS studies (0.82; 95%C.I. 0.63 to 1). Exploratory analyses show that sham response is associated with refractoriness and with the use of rTMS as an add-on therapy, but not with age, gender and sham method utilized. CONCLUSIONS/SIGNIFICANCE: We confirmed that placebo response in MDD is large regardless of the intervention and is associated with depression refractoriness and treatment combination (add-on rTMS studies). The magnitude of the placebo response seems to be related with study population and study design rather than the intervention itself. PMID- 19293927 TI - S9, a novel anticancer agent, exerts its anti-proliferative activity by interfering with both PI3K-Akt-mTOR signaling and microtubule cytoskeleton. AB - BACKGROUND: Deregulation of the phosphatidylinositol 3-kinases (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway plays a central role in tumor formation and progression, providing validated targets for cancer therapy. S9, a hybrid of alpha-methylene-gamma-lactone and 2-phenyl indole compound, possessed potent activity against this pathway. METHODOLOGY/PRINCIPAL FINDINGS: Effects of S9 on PI3K-Akt-mTOR pathway were determined by Western blot, immunofluorescence staining and in vitro kinas assay. The interactions between tubulin and S9 were investigated by polymerization assay, CD, and SPR assay. The potential binding modes between S9 and PI3K, mTOR or tubulin were analyzed by molecular modeling. Anti-tumor activity of S9 was evaluated in tumor cells and in nude mice bearing human cancer xenografts. S9 abrogated EGF-activated PI3K-Akt mTOR signaling cascade and Akt translocation to cellular membrane in human tumor cells. S9 possessed inhibitory activity against both PI3K and mTOR with little effect on other tested 30 kinases. S9 also completely impeded hyper phosphorylation of Akt as a feedback of inhibition of mTOR by rapamycin. S9 unexpectedly arrested cells in M phase other than G1 phase, which was distinct from compounds targeting PI3K-Akt-mTOR pathway. Further study revealed that S9 inhibited tubulin polymerization via binding to colchicine-binding site of tubulin and resulted in microtubule disturbance. Molecular modeling indicated that S9 could potentially bind to the kinase domains of PI3K p110alpha subunit and mTOR, and shared similar hydrophobic interactions with colchicines in the complex with tubulin. Moreover, S9 induced rapid apoptosis in tumor cell, which might reflect a synergistic cooperation between blockade of both PI3-Akt-mTOR signaling and tubulin cytoskeleton. Finally, S9 displayed potent antiproliferative activity in a panel of tumor cells originated from different tissue types including drug-resistant cells and in nude mice bearing human tumor xenografts. CONCLUSIONS/SIGNIFICANCE: Taken together, S9 targets both PI3K-Akt mTOR signaling and microtubule cytoskeleton, which combinatorially contributes its antitumor activity and provides new clues for anticancer drug design and development. PMID- 19293928 TI - Detection of CWD prions in urine and saliva of deer by transgenic mouse bioassay. AB - Chronic wasting disease (CWD) is a prion disease affecting captive and free ranging cervids (e.g. deer, elk, and moose). The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role. Here we report the presence of infectious prions in the urine and saliva of deer with chronic wasting disease (CWD). Prion infectivity was detected by bioassay of concentrated, dialyzed urine and saliva in transgenic mice expressing the cervid PrP gene (Tg[CerPrP] mice). In addition, PrP(CWD) was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA). The concentration of abnormal prion protein in bodily fluids was very low, as indicated by: undetectable PrP(CWD) levels by traditional assays (western blot, ELISA) and prolonged incubation periods and incomplete TSE attack rates in inoculated Tg(CerPrP) mice (373(+/-)3 days in 2 of 9 urine-inoculated mice and 342(+/-)109 days in 8 of 9 saliva-inoculated mice). These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections. PMID- 19293929 TI - Collagen-based mechanical anisotropy of the tectorial membrane: implications for inter-row coupling of outer hair cell bundles. AB - BACKGROUND: The tectorial membrane (TM) in the mammalian cochlea displays anisotropy, where mechanical or structural properties differ along varying directions. The anisotropy arises from the presence of collagen fibrils organized in fibers of approximately 1 microm diameter that run radially across the TM. Mechanical coupling between the TM and the sensory epithelia is required for normal hearing. However, the lack of a suitable technique to measure mechanical anisotropy at the microscale level has hindered understanding of the TM's precise role. METHODOLOGY/PRINCIPAL FINDINGS: Here we report values of the three elastic moduli that characterize the anisotropic mechanical properties of the TM. Our novel technique combined Atomic Force Microscopy (AFM), modeling, and optical tracking of microspheres to determine the elastic moduli. We found that the TM's large mechanical anisotropy results in a marked transmission of deformations along the direction that maximizes sensory cell excitation, whereas in the perpendicular direction the transmission is greatly reduced. CONCLUSIONS/SIGNIFICANCE: Computational results, based on our values of elastic moduli, suggest that the TM facilitates the directional cooperativity of sensory cells in the cochlea, and that mechanical properties of the TM are tuned to guarantee that the magnitude of sound-induced tip-link stretching remains similar along the length of the cochlea. Furthermore, we anticipate our assay to be a starting point for other studies of biological tissues that require directional functionality. PMID- 19293930 TI - Adult-young ratio, a major factor regulating social behaviour of young: a horse study. AB - BACKGROUND: Adults play an important role in regulating the social behaviour of young individuals. However, a few pioneer studies suggest that, more than the mere presence of adults, their proportions in social groups affect the social development of young. Here, we hypothesized that aggression rates and social cohesion were correlated to adult-young ratios. Our biological model was naturally-formed groups of Przewalski horses, Equus f. przewalskii, varying in composition. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the social interactions and spatial relationships of 12 one- and two-year-old Przewalski horses belonging to five families with adult-young ratios (AYR) ranging from 0.67 to 1.33. We found striking variations of aggression rates and spatial relationships related to the adult-young ratio: the lower this ratio, the more the young were aggressive, the more young and adults segregated and the tighter the young bonded to other young. CONCLUSION/SIGNIFICANCE: This is the first study demonstrating a correlation between adult-young ratios and aggression rates and social cohesion of young individuals in a naturalistic setting. The increase of aggression and the emergence of social segregation in groups with lower proportions of adults could reflect a related decrease of the influence of adults as regulators of the behaviour of young. This social regulation has both theoretical and practical implications for understanding the modalities of the influence of adults during ontogeny and for recommending optimal settings, as for instance, for schooling or animal group management. PMID- 19293931 TI - Inhibition of GSK3 phosphorylation of beta-catenin via phosphorylated PPPSPXS motifs of Wnt coreceptor LRP6. AB - The Wnt/beta-catenin signaling pathway plays essential roles in cell proliferation and differentiation, and deregulated beta-catenin protein levels lead to many types of human cancers. On activation by Wnt, the Wnt co-receptor LDL receptor related protein 6 (LRP6) is phosphorylated at multiple conserved intracellular PPPSPXS motifs by glycogen synthase kinase 3 (GSK3) and casein kinase 1 (CK1), resulting in recruitment of the scaffolding protein Axin to LRP6. As a result, beta-catenin phosphorylation by GSK3 is inhibited and beta-catenin protein is stabilized. However, how LRP6 phosphorylation and the ensuing LRP6 Axin interaction lead to the inhibition of beta-catenin phosphorylation by GSK3 is not fully understood. In this study, we reconstituted Axin-dependent beta catenin phosphorylation by GSK3 and CK1 in vitro using recombinant proteins, and found that the phosphorylated PPPSPXS peptides directly inhibit beta-catenin phosphorylation by GSK3 in a sequence and phosphorylation-dependent manner. This inhibitory effect of phosphorylated PPPSPXS motifs is direct and specific for GSK3 phosphorylation of beta-catenin at Ser33/Ser37/Thr41 but not for CK1 phosphorylation of beta-catenin at Ser45, and is independent of Axin function. We also show that a phosphorylated PPPSPXS peptide is able to activate Wnt/beta catenin signaling and to induce axis duplication in Xenopus embryos, presumably by inhibition of GSK3 in vivo. Based on these observations, we propose a working model that Axin recruitment to the phosphorylated LRP6 places GSK3 in the vicinity of multiple phosphorylated PPPSPXS motifs, which directly inhibit GSK3 phosphorylation of beta-catenin. This model provides a possible mechanism to account, in part, for inhibition of beta-catenin phosphorylation by Wnt-activated LRP6. PMID- 19293932 TI - Molecular basis of filamin A-FilGAP interaction and its impairment in congenital disorders associated with filamin A mutations. AB - BACKGROUND: Mutations in filamin A (FLNa), an essential cytoskeletal protein with multiple binding partners, cause developmental anomalies in humans. METHODOLOGY/PRINCIPAL FINDINGS: We determined the structure of the 23rd Ig repeat of FLNa (IgFLNa23) that interacts with FilGAP, a Rac-specific GTPase-activating protein and regulator of cell polarity and movement, and the effect of the three disease-related mutations on this interaction. A combination of NMR structural analysis and in silico modeling revealed the structural interface details between the C and D beta-strands of the IgFLNa23 and the C-terminal 32 residues of FilGAP. Mutagenesis of the predicted key interface residues confirmed the binding constraints between the two proteins. Specific loss-of-function FLNa constructs were generated and used to analyze the importance of the FLNa-FilGAP interaction in vivo. Point mutagenesis revealed that disruption of the FLNa-FilGAP interface perturbs cell spreading. FilGAP does not bind FLNa homologs FLNb or FLNc establishing the importance of this interaction to the human FLNa mutations. Tight complex formation requires dimerization of both partners and the correct alignment of the binding surfaces, which is promoted by a flexible hinge domain between repeats 23 and 24 of FLNa. FLNa mutations associated with human developmental anomalies disrupt the binding interaction and weaken the elasticity of FLNa/F-actin network under high mechanical stress. CONCLUSIONS/SIGNIFICANCE: Mutational analysis informed by structure can generate reagents for probing specific cellular interactions of FLNa. Disease-related FLNa mutations have demonstrable effects on FLNa function. PMID- 19293933 TI - I feel what you feel if you are similar to me. AB - Social interactions are influenced by the perception of others as similar or dissimilar to the self. Such judgements could depend on physical and semantic characteristics, such as membership in an ethnic or political group. In the present study we tested whether social representations of the self and of others could affect the perception of touch. To this aim, we assessed tactile perception on the face when subjects observed a face being touched by fingers. In different conditions we manipulated the identity of the shown face. In a first experiment, Caucasian and Maghrebian participants viewed a face belonging either to their own or to a different ethnic group; in a second experiment, Liberal and Conservative politically active participants viewed faces of politicians belonging to their own or to the opposite political party. The results showed that viewing a touched face most strongly enhanced the perception of touch on the observer's face when the observed face belonged to his/her own ethnic or political group. PMID- 19293934 TI - Genome-wide hypomethylation in head and neck cancer is more pronounced in HPV negative tumors and is associated with genomic instability. AB - Loss of genome-wide methylation is a common feature of cancer, and the degree of hypomethylation has been correlated with genomic instability. Global methylation of repetitive elements possibly arose as a defense mechanism against parasitic DNA elements, including retrotransposons and viral pathogens. Given the alterations of global methylation in both viral infection and cancer, we examined genome-wide methylation levels in head and neck squamous cell carcinoma (HNSCC), a cancer causally associated with human papilloma virus (HPV). We assayed global hypomethylation levels in 26 HNSCC samples, compared with their matched normal adjacent tissue, using Pyrosequencing-based methylation assays for LINE repeats. In addition, we examined cell lines derived from a variety of solid tumors for LINE and SINE (Alu) repeats. The degree of LINE and Alu hypomethylation varied among different cancer cell lines. There was only moderate correlation between LINE and Alu methylation levels, with the range of variation in methylation levels being greater for the LINE elements. LINE hypomethylation was more pronounced in HPV-negative than in HPV-positive tumors. Moreover, genomic instability, as measured by genome-wide loss-of-heterozygosity (LOH) single nucleotide polymorphism (SNP) analysis, was greater in HNSCC samples with more pronounced LINE hypomethylation. Global hypomethylation was variable in HNSCC. Its correlation with both HPV status and degree of LOH as a surrogate for genomic instability may reflect alternative oncogenic pathways in HPV-positive versus HPV negative tumors. PMID- 19293935 TI - Host genetic background strongly influences the response to influenza a virus infections. AB - The genetic make-up of the host has a major influence on its response to combat pathogens. For influenza A virus, several single gene mutations have been described which contribute to survival, the immune response and clearance of the pathogen by the host organism. Here, we have studied the influence of the genetic background to influenza A H1N1 (PR8) and H7N7 (SC35M) viruses. The seven inbred laboratory strains of mice analyzed exhibited different weight loss kinetics and survival rates after infection with PR8. Two strains in particular, DBA/2J and A/J, showed very high susceptibility to viral infections compared to all other strains. The LD(50) to the influenza virus PR8 in DBA/2J mice was more than 1000 fold lower than in C57BL/6J mice. High susceptibility in DBA/2J mice was also observed after infection with influenza strain SC35M. In addition, infected DBA/2J mice showed a higher viral load in their lungs, elevated expression of cytokines and chemokines, and a more severe and extended lung pathology compared to infected C57BL/6J mice. These findings indicate a major contribution of the genetic background of the host to influenza A virus infections. The overall response in highly susceptible DBA/2J mice resembled the pathology described for infections with the highly virulent influenza H1N1-1918 and newly emerged H5N1 viruses. PMID- 19293936 TI - Health and human rights education in U.S. schools of medicine and public health: current status and future challenges. AB - BACKGROUND: Despite increasing recognition of the importance of human rights in the protection and promotion of health, formal human rights education has been lacking in schools of medicine and public health. Our objectives were: 1) to determine the nature and extent of health and human rights (HHR) education among schools of medicine (SOMs) and public health (SPHs); 2) to identify perceived barriers to implementing HHR curricula; 3) to learn about deans' interests and attitudes toward HHR education, and; 4) to identify factors associated with offering HHR education. METHODS AND PRINCIPAL FINDINGS: We conducted a cross sectional survey among deans of all accredited allopathic SOMs and SPHs in the United States and Puerto Rico. Seventy-one percent of U.S. SOMs and SPHs responded. Thirty-seven percent of respondents indicated that their schools offered some form of HHR education. Main barriers to offering HHR education included competition for time, lack of qualified instructors and lack of funding. Among schools not offering HHR education, 35% of deans were interested in offering HHR education. Seventy-six percent of all deans believed that it was very important or important to offer HHR education. Multiple regression analysis revealed that deans' attitudes were the most important factor associated with offering any HHR education. CONCLUSION: Findings indicate that though a majority of deans of SOMs and SPHs believe that knowledge about human rights is important in health practice and support the inclusion of HHR studies in their schools, HHR education is lacking at most of their institutions. These results and the growing recognition of the critical interdependence between health and human rights indicate a need for SOMs and SPHs to work towards formal inclusion of HHR studies in their curricula, and that HHR competency requirements be considered to overcome barriers to its inclusion. PMID- 19293937 TI - The influence of recovery and training phases on body composition, peripheral vascular function and immune system of professional soccer players. AB - Professional soccer players have a lengthy playing season, throughout which high levels of physical stress are maintained. The following recuperation period, before starting the next pre-season training phase, is generally considered short but sufficient to allow a decrease in these stress levels and therefore a reduction in the propensity for injury or musculoskeletal tissue damage. We hypothesised that these physical extremes influence the body composition, blood flow, and endothelial/immune function, but that the recuperation may be insufficient to allow a reduction of tissue stress damage. Ten professional football players were examined at the end of the playing season, at the end of the season intermission, and after the next pre-season endurance training. Peripheral blood flow and body composition were assessed using venous occlusion plethysmography and DEXA scanning respectively. In addition, selected inflammatory and immune parameters were analysed from blood samples. Following the recuperation period a significant decrease of lean body mass from 74.4+/-4.2 kg to 72.2+/-3.9 kg was observed, but an increase of fat mass from 10.3+/-5.6 kg to 11.1+/-5.4 kg, almost completely reversed the changes seen in the pre-season training phase. Remarkably, both resting and post-ischemic blood flow (7.3+/-3.4 and 26.0+/-6.3 ml/100 ml/min) respectively, were strongly reduced during the playing and training stress phases, but both parameters increased to normal levels (9.0+/-2.7 and 33.9+/-7.6 ml/100 ml/min) during the season intermission. Recovery was also characterized by rising levels of serum creatinine, granulocytes count, total IL-8, serum nitrate, ferritin, and bilirubin. These data suggest a compensated hypo-perfusion of muscle during the playing season, followed by an intramuscular ischemia/reperfusion syndrome during the recovery phase that is associated with muscle protein turnover and inflammatory endothelial reaction, as demonstrated by iNOS and HO-1 activation, as well as IL 8 release. The data provided from this study suggest that the immune system is not able to function fully during periods of high physical stress. The implications of this study are that recuperation should be carefully monitored in athletes who undergo intensive training over extended periods, but that these parameters may also prove useful for determining an individual's risk of tissue stress and possibly their susceptibility to progressive tissue damage or injury. PMID- 19293938 TI - Enterohemorrhagic Escherichia coli O157:H7 gene expression profiling in response to growth in the presence of host epithelia. AB - BACKGROUND: The pathogenesis of enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection is attributed to virulence factors encoded on multiple pathogenicity islands. Previous studies have shown that EHEC O157:H7 modulates host cell signal transduction cascades, independent of toxins and rearrangement of the cytoskeleton. However, the virulence factors and mechanisms responsible for EHEC mediated subversion of signal transduction remain to be determined. Therefore, the purpose of this study was to first identify differentially regulated genes in response to EHEC O157:H7 grown in the presence of epithelial cells, compared to growth in the absence of epithelial cells (that is, growth in minimal essential tissue culture medium alone, minimal essential tissue culture medium in the presence of 5% CO(2), and Penassay broth alone) and, second, to identify EHEC virulence factors responsible for pathogen modulation of host cell signal transduction. METHODOLOGY/PRINCIPAL FINDINGS: Overnight cultures of EHEC O157:H7 were incubated for 6 hr at 37 degrees C in the presence or absence of confluent epithelial (HEp-2) cells. Total RNA was then extracted and used for microarray analyses (Affymetrix E. coli Genome 2.0 gene chips). Relative to bacteria grown in each of the other conditions, EHEC O157:H7 cultured in the presence of cultured epithelial cells displayed a distinct gene-expression profile. A 2.0 fold increase in the expression of 71 genes and a 2.0-fold decrease in expression of 60 other genes were identified in EHEC O157:H7 grown in the presence of epithelial cells, compared to bacteria grown in media alone. CONCLUSION/SIGNIFICANCE: Microarray analyses and gene deletion identified a protease on O-island 50, gene Z1787, as a potential virulence factor responsible for mediating EHEC inhibition of the interferon (IFN)-gamma-Jak1,2-STAT-1 signal transduction cascade. Up-regulated genes provide novel targets for use in developing strategies to interrupt the infectious process. PMID- 19293939 TI - Effect of cigarette smoke extract on dendritic cells and their impact on T-cell proliferation. AB - Chronic obstructive pulmonary disease (COPD) is characterized by chronic airway inflammation. Cigarette smoke has been considered a major player in the pathogenesis of COPD. The inflamed airways of COPD patients contain several inflammatory cells including neutrophils, macrophages,T lymphocytes, and dendritic cells (DCs). The relative contributions of these various inflammatory cells to airway injury and remodeling are not well documented. In particular, the potential role of DCs as mediators of inflammation in the smoker's airways and COPD patients is poorly understood. In the current study we analyzed the effects of cigarette smoke extract on mouse bone marrow derived DC and the production of chemokines and cytokines were studied. In addition, we assessed CSE-induced changes in cDC function in the mixed lymphocyte reaction (MLR) examining CD4+ and CD8+ T cell proliferation. Cigarette smoke extract induces the release of the chemokines CCL3 and CXCL2 (but not cytokines), via the generation of reactive oxygen species (ROS). In a mixed-leukocyte reaction assay, cigarette smoke-primed DCs potentiate CD8(+)T cell proliferation via CCL3. In contrast, proliferation of CD4(+)T cells is suppressed via an unknown mechanism. The cigarette smoke-induced release of CCL3 and CXCL2 by DCs may contribute to the influx of CD8(+)T cells and neutrophils into the airways, respectively. PMID- 19293940 TI - Uncontrolled airway inflammation in lung disease represents a defect in counter regulatory signaling. AB - Counter-regulatory lipid mediators are generated during airway inflammation to promote resolution. Defects in the production of these lipid mediators have now been associated with several diseases of persistent airway inflammation. Lipoxins are the lead members of this class of anti-inflammatory and proresolving chemical mediators. Recently, several new families of fatty acid-derived counter regulatory mediators have been discovered, including the resolvins and protectins. Diminished formation of these endogenous protective signals would interrupt the natural resolution of inflammation. Here, we review how rapidly emerging information on lipoxins, resolvins and protectins is providing new insights into the pathophysiology of chronic airway inflammation in several common illnesses, including asthma. PMID- 19293941 TI - Stigma management? The links between enacted stigma and teen pregnancy trends among gay, lesbian, and bisexual students in British Columbia. AB - Over the past decade, several large-scale school-based studies of adolescents in Canada and the U.S. have documented health disparities for lesbian, gay and bisexual teens compared to their heterosexual peers, such as higher rates of suicide attempts, homelessness, and substance use. Many of these disparities have been linked to "enacted stigma," or the higher rates of harassment, discrimination, and sexual or physical violence that sexual minority youth experience at home, at school, and in the community. An unexpected health disparity for lesbia n, gay and bisexual youth is their significantly higher risk of teen pregnancy involvement (between two and seven times the rate of their heterosexual peers), especially in light of declining trends in teen pregnancy across North America since the early 1990s. What is behind this higher risk? Is it getting better or worse? Using the province-wide cluster-stratified British Columbia Adolescent Health Surveys from 1992, 1998, and 2003, this paper explores the trends in pregnancy involvement, related sexual behaviours, and exposure to forms of enacted stigma that may help explain this particular health disparity for gay, lesbian and bisexual youth in Canada. PMID- 19293944 TI - Multivalent HA DNA vaccination protects against highly pathogenic H5N1 avian influenza infection in chickens and mice. AB - BACKGROUND: Sustained outbreaks of highly pathogenic avian influenza (HPAI) H5N1 in avian species increase the risk of reassortment and adaptation to humans. The ability to contain its spread in chickens would reduce this threat and help maintain the capacity for egg-based vaccine production. While vaccines offer the potential to control avian disease, a major concern of current vaccines is their potency and inability to protect against evolving avian influenza viruses. METHODOLOGY / PRINCIPAL FINDINGS: The ability of DNA vaccines encoding hemagglutinin (HA) proteins from different HPAI H5N1 serotypes was evaluated for its ability to elicit neutralizing antibodies and to protect against homologous and heterologous HPAI H5N1 strain challenge in mice and chickens after DNA immunization by needle and syringe or with a pressure injection device. These vaccines elicited antibodies that neutralized multiple strains of HPAI H5N1 when given in combinations containing up to 10 HAs. The response was dose-dependent, and breadth was determined by the choice of the influenza virus HA in the vaccine. Monovalent and trivalent HA vaccines were tested first in mice and conferred protection against lethal H5N1 A/Vietnam/1203/2004 challenge 68 weeks after vaccination. In chickens, protection was observed against heterologous strains of HPAI H5N1 after vaccination with a trivalent H5 serotype DNA vaccine with doses as low as 5 microg DNA given twice either by intramuscular needle injection or with a needle-free device. CONCLUSIONS/SIGNIFICANCE: DNA vaccines offer a generic approach to influenza virus immunization applicable to multiple animal species. In addition, the ability to substitute plasmids encoding different strains enables rapid adaptation of the vaccine to newly evolving field isolates. PMID- 19293945 TI - A human protein interaction network shows conservation of aging processes between human and invertebrate species. AB - We have mapped a protein interaction network of human homologs of proteins that modify longevity in invertebrate species. This network is derived from a proteome scale human protein interaction Core Network generated through unbiased high throughput yeast two-hybrid searches. The longevity network is composed of 175 human homologs of proteins known to confer increased longevity through loss of function in yeast, nematode, or fly, and 2,163 additional human proteins that interact with these homologs. Overall, the network consists of 3,271 binary interactions among 2,338 unique proteins. A comparison of the average node degree of the human longevity homologs with random sets of proteins in the Core Network indicates that human homologs of longevity proteins are highly connected hubs with a mean node degree of 18.8 partners. Shortest path length analysis shows that proteins in this network are significantly more connected than would be expected by chance. To examine the relationship of this network to human aging phenotypes, we compared the genes encoding longevity network proteins to genes known to be changed transcriptionally during aging in human muscle. In the case of both the longevity protein homologs and their interactors, we observed enrichments for differentially expressed genes in the network. To determine whether homologs of human longevity interacting proteins can modulate life span in invertebrates, homologs of 18 human FRAP1 interacting proteins showing significant changes in human aging muscle were tested for effects on nematode life span using RNAi. Of 18 genes tested, 33% extended life span when knocked down in Caenorhabditis elegans. These observations indicate that a broad class of longevity genes identified in invertebrate models of aging have relevance to human aging. They also indicate that the longevity protein interaction network presented here is enriched for novel conserved longevity proteins. PMID- 19293946 TI - Alignment and prediction of cis-regulatory modules based on a probabilistic model of evolution. AB - Cross-species comparison has emerged as a powerful paradigm for predicting cis regulatory modules (CRMs) and understanding their evolution. The comparison requires reliable sequence alignment, which remains a challenging task for less conserved noncoding sequences. Furthermore, the existing models of DNA sequence evolution generally do not explicitly treat the special properties of CRM sequences. To address these limitations, we propose a model of CRM evolution that captures different modes of evolution of functional transcription factor binding sites (TFBSs) and the background sequences. A particularly novel aspect of our work is a probabilistic model of gains and losses of TFBSs, a process being recognized as an important part of regulatory sequence evolution. We present a computational framework that uses this model to solve the problems of CRM alignment and prediction. Our alignment method is similar to existing methods of statistical alignment but uses the conserved binding sites to improve alignment. Our CRM prediction method deals with the inherent uncertainties of binding site annotations and sequence alignment in a probabilistic framework. In simulated as well as real data, we demonstrate that our program is able to improve both alignment and prediction of CRM sequences over several state-of-the-art methods. Finally, we used alignments produced by our program to study binding site conservation in genome-wide binding data of key transcription factors in the Drosophila blastoderm, with two intriguing results: (i) the factor-bound sequences are under strong evolutionary constraints even if their neighboring genes are not expressed in the blastoderm and (ii) binding sites in distal bound sequences (relative to transcription start sites) tend to be more conserved than those in proximal regions. Our approach is implemented as software, EMMA (Evolutionary Model-based cis-regulatory Module Analysis), ready to be applied in a broad biological context. PMID- 19293947 TI - Physical health care in persons with severe mental illness: a public health and ethical priority. PMID- 19293948 TI - Enhancing research and treatment of mental disorders with dimensional concepts: toward DSM-V and ICD-11. AB - The current versions of the DSM (DSM-IV-TR) and ICD (ICD-10) describe all mental disorders as polythetic-categorical concepts. Lists of symptoms are presented, and diagnostic category labels are assigned to patients based on observing specific patterns of symptoms. A number of notable conceptual problems emerge when using this strictly categorical system in research and in the clinic. When thorough structured diagnostic interviews are used, typical patients meet criteria for more than one specific diagnosis (a phenomenon termed "comorbidity"). In addition, groups of patients with the same putative categorical label are often heterogeneous with respect to key clinical features, such as severity and prognosis, and patients with symptomatology below diagnostic thresholds are often significantly impaired. Although categorical concepts will always be essential in official nosologies (e.g., in providing diagnostic labels for reimbursement purposes), many of the conceptual problems of a strictly categorical diagnostic system can be overcome by enhancing official nosologies with dimensional concepts. Specific dimensional approaches and directions that may be considered for upcoming revisions of both the DSM and ICD are discussed. PMID- 19293949 TI - Relationship problems and the DSM:needed improvements and suggested solutions. AB - Relational problems are clinically significant behavioral or psychological syndromes or patterns that occur between or among individuals and that are associated with present distress or disability or with a significant increased risk of suffering death, pain, disability, or an important loss of freedom. Relational problems (e.g., partner relational problems, partner abuse, child maltreatment) are included as Axis I disorders in the DSM-IV as V-codes (i.e., "Other conditions that may be a focus of clinical attention"). However, there are no criteria provided in the DSM-IV for these codes. In this article, we briefly review literature that incontrovertibly documents both relational problems' syndromes/patterns and their serious sequelae. We then review a series of studies that provide evidence of content validity and inter-rater agreement for criteria to determine presence versus absence of relational problems. The most studied subset of relational problem criteria, those for partner and child maltreatment, have been shown to have remarkably high levels of reliability when disseminated broadly in the field (kappa = .66-.89), at agreement levels never reached by DSM diagnoses for individuals. We conclude by arguing that science, service, families, individuals, and the DSM itself, would be well served to include diagnostic criteria for relational problems and to consider the various options for placement of relational problems/processes in the DSM-V. PMID- 19293950 TI - Metabolic syndrome in people with schizophrenia: a review. AB - Metabolic syndrome and other cardiovascular risk factors are highly prevalent in people with schizophrenia. Patients are at risk for premature mortality and overall have limited access to physical health care. In part these cardio metabolic risk factors are attributable to unhealthy lifestyle, including poor diet and sedentary behaviour. But over recent years it has become apparent that antipsychotic agents can have a negative impact on some of the modifiable risk factors. The psychiatrist needs to be aware of the potential metabolic side effects of antipsychotic medication and to include them in the risk/benefit assessment when choosing a specific antipsychotic. He should also be responsible for the implementation of the necessary screening assessments and referral for treatment of any physical illness. Multidisciplinary assessment of psychiatric and medical conditions is needed. The somatic treatments offered to people with severe and enduring mental illness should be at par with general health care in the non-psychiatrically ill population. PMID- 19293951 TI - Effectiveness as an outcome measure for treatment trials in psychiatry. AB - There is at present some confusion about the relative value of clinical trials performed to investigate efficacy vs. those designed to investigate effectiveness. This is particularly challenging when studies performed as experiments for regulators by companies are used to shape and inform clinical practice, especially if studies conducted under more real life conditions fail to support predicted benefits. We review the field in relation to the new antipsychotics, in particular. Other indications, including mood disorders, which are also briefly touched upon, have so far received less definitive attention, but are likely to encounter the same difficulties. We conclude that, where the results of efficacy trials are positive and an effectiveness trial is negative, one should not necessarily prefer the effectiveness trial - it may simply have failed. Where efficacy trials and effectiveness trials point to similar conclusions, then the findings are mutually supportive. PMID- 19293952 TI - Clinical trial design: horses for courses. PMID- 19293953 TI - Much ado about small differences. PMID- 19293954 TI - The silver lining of recent effectiveness trials. PMID- 19293955 TI - Do "real world" studies on antipsychotics tell us the real truth? PMID- 19293956 TI - The quest for a meaningful evidence base in psychiatry. PMID- 19293957 TI - The role of efficacy and effectiveness trials. PMID- 19293958 TI - Social functioning and quality of life as measures of effectiveness in the treatment of schizophrenia. PMID- 19293959 TI - Indexation of psychiatric journals from low- and middle-income countries: a survey and a case study. AB - The objective of this cross-sectional study was to determine the prevalence of migraine headache among depressed Latino adolescents of Mexican American origin. This is, to the best of our knowledge, the first study of the prevalence of migraine among depressed adolescents of any ethnic/racial background. In a mental health clinic for the indigent, 132 consecutive Latino adolescents fulfilling the DSM-IV criteria for major depressive episode were compared with a sample of adolescents with other mental disorders. Logistic regression was used to test for associations and control for confounding effects. The prevalence of migraine headache among depressed adolescents was 6 times greater than that of the comparison patients (OR = 5.98, z = 2.35, p = 0.019). This finding is consistent with previously published reports involving adult samples, in which the prevalence of migraine was found to exceed that in the general population. However, contrary to what we previously found in Latino adults, the prevalence of migraine was not higher in bipolar than in unipolar adolescents. PMID- 19293960 TI - Mental health policies on reporting child sexual abuse and physician-patient sexual relationships. AB - The reporting of child sexual abuse (CSA) and physician-patient sexual relationships (PPSR) are currently the focus of professional, legal and media attention in several countries. This paper briefly reviews mental health policies on these issues and reports on a WPA survey of them. While the WPA Madrid Declaration permits breaching confidentiality for mandatory reporting of CSA and clearly prohibits PPSR, it is not known how or to what extent these policies are implemented in WPA Member Societies' countries. It is also not known whether policies or laws exist on these topics nationally or to what extent psychiatrists and the public are aware of them. Representatives of WPA Member Societies were e mailed a survey about issues pertaining to CSA and PPSR. Fifty-one percent of 109 countries replied. All reporting countries had laws or policies regarding the reporting of CSA, but this was often voluntary (63%) and without protection for reporting psychiatrists either by law (29%) or by Member Societies (27%). A substantial number of psychiatric leaders did not know the law (27%) or their Society's policy (11%) on these matters. With respect to PPSR, some reporting countries lacked laws or policies about PPSR with current (17%) or past (56%) patients. Fewer than half of responding representatives believed that their Society's members or the public were well informed about the laws and policies pertaining to CSA or PPSR. There is clearly a wide range of laws, policies and practices about CSA and PPSR in WPA Member Societies' countries. There is a need in some countries for laws or supplemental policies to facilitate the protection of vulnerable child and adult patients through clear, mandatory reporting policies for CSA and PPSR. Mechanisms to protect and support reporting psychiatrists should also be developed where they do not already exist. There is also a need in some countries to develop strategies to improve the education of psychiatrists, trainees, and the public on these issues. PMID- 19293961 TI - Community mental health care in the Asia-Pacific region: using current best practice models to inform future policy. AB - The reporting of child sexual abuse (CSA) and physician-patient sexual relationships (PPSR) are currently the focus of professional, legal and media attention in several countries. This paper briefly reviews mental health policies on these issues and reports on a WPA survey of them. While the WPA Madrid Declaration permits breaching confidentiality for mandatory reporting of CSA and clearly prohibits PPSR, it is not known how or to what extent these policies are implemented in WPA Member Societies' countries. It is also not known whether policies or laws exist on these topics nationally or to what extent psychiatrists and the public are aware of them. Representatives of WPA Member Societies were e mailed a survey about issues pertaining to CSA and PPSR. Fifty-one percent of 109 countries replied. All reporting countries had laws or policies regarding the reporting of CSA, but this was often voluntary (63%) and without protection for reporting psychiatrists either by law (29%) or by Member Societies (27%). A substantial number of psychiatric leaders did not know the law (27%) or their Society's policy (11%) on these matters. With respect to PPSR, some reporting countries lacked laws or policies about PPSR with current (17%) or past (56%) patients. Fewer than half of responding representatives believed that their Society's members or the public were well informed about the laws and policies pertaining to CSA or PPSR. There is clearly a wide range of laws, policies and practices about CSA and PPSR in WPA Member Societies' countries. There is a need in some countries for laws or supplemental policies to facilitate the protection of vulnerable child and adult patients through clear, mandatory reporting policies for CSA and PPSR. Mechanisms to protect and support reporting psychiatrists should also be developed where they do not already exist. There is also a need in some countries to develop strategies to improve the education of psychiatrists, trainees, and the public on these issues. PMID- 19293963 TI - Letter to the editor. PMID- 19293962 TI - Embodiment and schizophrenia. AB - The essential feature of schizophrenic existence is its being disembodied. This is the feature that unifies the varied dimensions of that existence. The disembodiment of the self, of the self-object relation and of interpersonal relationships all lead to a kind of world in which the schizophrenic person lives and behaves like a soulless body or a disembodied spirit. PMID- 19293964 TI - The WPA General Assembly in Prague and the new WPA leadership. PMID- 19293965 TI - The WPA International Congress "Treatment in Psychiatry A New Update" (Florence, April 1-4, 2009). PMID- 19293966 TI - Assessing life expectancy: our continuing challenge. PMID- 19293967 TI - The importance of feedback. PMID- 19293968 TI - Note of gratitude. PMID- 19293969 TI - Treating patients with postoperative stress urinary incontinence. PMID- 19293970 TI - Survival after radical prostatectomy and radiotherapy for prostate cancer: a population-based study. AB - OBJECTIVE: Based on the natural history of localized prostate cancer, the life expectancy (LE) of men treated with either radical prostatectomy (RP) or definitive external-beam radiotherapy (EBRT) should exceed 10 years. To test this hypothesis, we examined overall survival rates after RP or EBRT in a contemporary population-based cohort. METHODS: Within a population-based cohort we assessed crude survival in 17 570 men diagnosed with prostate cancer who were either treated with RP (n = 9678) or definitive EBRT (n = 7892) between 1989 and 2000. Age and Charlson Comorbidity Index (CCI) score at treatment represented covariates. In order to control for prostate cancer-related mortality, we repeated analyses for 9131 men who did not receive any secondary treatment for prostate cancer. RESULTS: In the entire cohort, the actuarial 10-year survival probability after RP was 75.3%, versus 36.7% after EBRT (p < 0.001). In those who did not receive any secondary treatment, the actuarial 10-year survival probability after RP was 81.1%, versus 30.4% after EBRT (p < 0.001). In multivariate Cox regression models, EBRT was associated with a 2.8-fold (p < 0.001) and 3.9-fold (p < 0.001) higher risk of mortality in the entire cohort and in the cohort without secondary treatment, respectively. Increased CCI score and increased age were also associated with a higher risk of mortality (p < 0.001). CONCLUSION: Some men treated with EBRT and, to a lesser extent, those treated with RP may have insufficient LE to warrant therapy with curative intent. More stringent selection criteria are necessary to avoid overtreatment. PMID- 19293971 TI - Task-specific bench model training versus basic laparoscopic skills training for laparoscopic radical prostatectomy: a randomized controlled study. AB - BACKGROUND: Performing a laparoscopic urethrovesical anastomosis (LUA) after a radical prostatectomy is technically challenging for the novice laparoscopic surgeon. We developed a low-fidelity urethrovesical model (UVM) to allow a urologist to practise this critical step. The aim of our study was to compare the effect of task-specific bench model training (anastomotic suturing on the UVM) with that of basic laparoscopic suturing on intracorporeal urethrovesical anastomosis performance. METHODS: We recruited 28 senior surgical residents, fellows or staff surgeons for this prospective, single-blinded, randomized controlled study. We randomly assigned participants to an intervention group practising LUA on the UVM or to a control group practising basic laparoscopic suturing and knot-tying on a foam pad. After practising, we videotaped participants performing 5 intra-corporeal interrupted sutures on a foam pad and a LUA on the UVM. A blinded expert scored the videotaped performance using a laparoscopic suturing checklist (CL) and a global rating scale (GRS), and timed the performance. RESULTS: On the foam pad suturing task, the group that trained on the UVM had significantly higher CL scores (10.9 v. 8.1, p = 0.017). On the LUA task, the group that trained on the UVM had significantly higher CL scores (10.9 v. 8.1, p = 0.017), GRS (29.6 v. 22.8, p = 0.005) and shorter times (27.6 v. 38.3 min, p = 0.004) than the control group. CONCLUSION: Our task-specific bench model was shown to be superior to basic laparoscopic suturing drills on a foam pad. PMID- 19293972 TI - Improving laparoscopic suturing skills. PMID- 19293973 TI - Hereditary renal cell carcinoma associated with von Hippel-Lindau disease: a description of a Nova Scotia cohort. AB - BACKGROUND: von Hippel-Lindau (VHL) disease is an autosomal dominant condition characterized by the development of benign and malignant tumours, including cases of renal cell carcinoma (RCC). Early detection of RCC through routine surveillance can lead to decreased morbidity and mortality. Data on the number of patients in Nova Scotia (NS) who have VHL disease, disease manifestations and the frequency and mode of the surveillance have not previously been collected or reported. This project was designed to obtain that information. METHODS: The number and management of patients with VHL disease was determined by multiple sources: the Maritime Medical Genetics Service, patient charts, and pathology, radiology and laboratory data. The actual surveillance being performed was compared with that recommended in the literature. RESULTS: Twenty-one patients from 11 families in NS were identified. Manifestations included cases of RCC (31.6%), central nervous system (CNS) hemangioblastoma (73.7%), retinal hemangioma (47.4%), renal cyst (47.4%) and pheochromocytoma (10.5%). Of the 6 patients with RCC, 4 had bilateral tumours, 2 required kidney transplants and 1 developed metastatic disease. Routine surveillance was being done for the CNS in 62.5% of patients, retina in 47.4%, abdomen in 43.8% and urine catecholamines in only 10.5%. Only 1 of the 6 patients who developed RCC was undergoing routine abdominal imaging. Surveillance investigations were ordered by a number of different specialists. CONCLUSION: Patients with VHL disease in NS have a number of manifestations associated with their disease, including RCC, in a similar frequency to that reported in the literature. The surveillance of these patients is suboptimal in frequency and coordination. von Hippel-Lindau disease is a complex condition that requires a coordinated approach to care to ensure proper surveillance and treatment. Our study highlights current deficiencies and offers an enormous opportunity for improvement. PMID- 19293974 TI - Penile rehabilitation following treatment for prostate cancer: an analysis of the current state of the art. AB - Despite recent advances in surgical technique using laparoscopic and robotic approaches for the management of early organ-confined prostate cancer, most contemporary reports demonstrate significant rates of erectile dysfunction comparable to standard open approaches. Controversy remains related to many of the pre-and postoperative management strategies, including agents to enhance nerve recovery, erectogenic drugs, antioxidants, vasoactive injectables, vacuum erection devices and nerve grafting procedures. Additionally, the optimal timing of these interventions and their duration, dose, frequency and outcome thresholds remain ill-defined. In our paper, we provide a comprehensive literature review involving both the basic and clinical data surrounding rehabilitative approaches. PMID- 19293975 TI - Optimizing sexual function outcomes after radical prostatectomy. PMID- 19293976 TI - Should penile rehabilitation become the norm following radical prostatectomy? PMID- 19293978 TI - Dr. Al Shaiji and Dr. Brock's rebuttal. PMID- 19293977 TI - Penile rehabilitation should not be the norm for patients post-radical prostatectomy. PMID- 19293979 TI - Dr. Lee's rebuttal. PMID- 19293980 TI - Recurrent idiopathic high-flow priapism treated with selective arterial embolization after repeated initial treatments for low-flow priapism. AB - High-flow priapism is most often seen following perineal and penile trauma. We report the case of a 32-year-old man who presented with recurrent idiopathic priapism initially treated as low-flow priapism on 8 previous emergency department visits. Pelvic angiography revealed an abnormal communication between the left cavernosal artery and the left corpus cavernosum and led to the diagnosis of high-flow priapism. Treatment involved embolization of the left common penile artery, which resulted in successful resolution of the recurrent priapism. Our patient's case highlights the importance of an appropriate work-up, including imaging, to distinguish high- and low-flow priapism and to provide appropriate care. PMID- 19293981 TI - Synchronous ipsilateral renal cell carcinoma and urothelial carcinoma of the renal pelvis. AB - Renal cell carcinoma (RCC) and urothelial carcinoma of the upper urinary tract are not uncommon urological malignancies. Their simultaneous occurrence in a patient is, however, extraordinarily rare. We report the case of a patient who underwent laparoscopic nephrectomy for suspected RCC. Preoperative imaging was suspicious for renal pelvic involvement, which was confirmed upon bivalving the fresh specimen at the time of surgery, with the discovery of a separate urothelium-based lesion. We discuss this rare occurrence and our management approach. PMID- 19293982 TI - Sports and the solitary kidney: what parents of a young child with a solitary kidney should know. PMID- 19293983 TI - Investigation and management of antenatally detected hydronephrosis. PMID- 19293984 TI - Follow-up guidelines after radical or partial nephrectomy for localized and locally advanced renal cell carcinoma. PMID- 19293985 TI - Canadian guidelines for the management of asymptomatic microscopic hematuria in adults. PMID- 19293987 TI - Urology meetings of interest. PMID- 19293986 TI - Diagnosis of interstitial cystitis June 2007. PMID- 19293989 TI - Phenotype analysis and quantification of proliferating cells in the cortical gray matter of the adult rat. AB - In intact adult mammalian brains, there are two neurogenic regions: the subependymal zone and the subgranular layer of the hippocampus. Even outside these regions, small numbers of proliferating precursors do exist. Many studies suggest that the majority of these are oligodendrocyte precursors that express NG2, a chondroitin sulfate proteoglycan, and most of the residual proliferating cells seem to be endothelial cells. However, it is still unclear whether NG2 immunonegative proliferating precursors are present, because previous studies have neglected their possible existence. In this study, we systematically analyzed the phenotypes of the proliferating cells in the intact adult rat cortical gray matter. We improved our techniques and carefully characterized the proliferating cells, because there were several problems with identifying and quantifying the proliferating cells: the detection of NG2-expressing cells was dependent on the fixation condition; there were residual proliferating leukocytes in the blood vessels; and two anti-NG2 antibodies gave rise to different staining patterns. Moreover, we used two methods, BrdU and Ki67 immunostaining, to quantify the proliferating cells. Our results strongly suggest that in the intact adult cerebral cortical gray matter, there were only two types of proliferating cells: the majority were NG2-expressing cells, including pericytes, and the rest were endothelial cells. PMID- 19293990 TI - Localization of reversion-induced LIM protein (RIL) in the rat central nervous system. AB - Reversion-induced LIM protein (RIL) is a member of the ALP (actinin-associated LIM protein) subfamily of the PDZ/LIM protein family. RIL serves as an adaptor protein and seems to regulate cytoskeletons. Immunoblotting suggested that RIL is concentrated in the astrocytes in the central nervous system. We then examined the expression and localization of RIL in the rat central nervous system and compared it with that of water channel aquaporin 4 (AQP4). RIL was concentrated in the cells of ependyma lining the ventricles in the brain and the central canal in the spinal cord. In most parts of the central nervous system, RIL was expressed in the astrocytes that expressed AQP4. Double-labeling studies showed that RIL was concentrated in the cytoplasm of astrocytes where glial fibrillary acidic protein was enriched as well as in the AQP4-enriched regions such as the endfeet or glia limitans. RIL was also present in some neurons such as Purkinje cells in the cerebellum and some neurons in the brain stem. Differential expression of RIL suggests that it may be involved in the regulation of the central nervous system. PMID- 19293991 TI - Finding Alu in primate genomes with AF-1. AB - Repetitive sequences occupy more than 40% of the human genome which is much larger compared to the 2% occupied by the coding DNA. Amongst these Alu elements are the second largest class of repeats, occupying nearly 10% of the whole genome. Alus have been implicated in many genomic processes, sometimes giving rise to aberrations while many times playing as silent player in genomic and regulatory evolution. Here we present a web server, AF1, exclusively developed for finding Alu like elements. Besides alignment based methodology, this server utilizes probabilistic scanning to find more diverged elements and employs a more precise way of element classification based on unequal weighting of sequence through sequence encoding. AVAILABILITY: AF1 is freely available at http://software.iiar.res.in/af1/. The standalone is also available for download. PMID- 19293992 TI - BrainSnail: a dynamic information display system for the sciences. AB - Scientific reference management has become crucial in rapidly expanding fields of biology. Many of the reference management systems currently employed are reference centric and not object/process focused. BrainSnail is a reference management/knowledge representation application that tries to bridge disconnect between subject and reference in the fields of neuropharmacology, neuroanatomy and neurophysiology. BrainSnail has been developed with considering both individual researcher and research group efforts. PMID- 19293993 TI - Extract-SAGE: an integrated platform for cross-analysis and GA-based selection of SAGE data. AB - Serial analysis of gene expression (SAGE) is a powerful quantification technique for gene expression data. The huge amount of tag data in SAGE libraries of samples is difficult to analyze with current SAGE analysis tools. Data is often not provided in a biologically significant way for cross-analysis and comparison, thus limiting its application. Hence, an integrated software platform that can perform such a complex task is required. Here, we implement set theory for cross-analyzing gene expression data among different SAGE libraries of tissue sources; up- or down-regulated tissue-specific tags can be identified computationally. Extract-SAGE employs a genetic algorithm (GA) to reduce the number of genes among the SAGE libraries. Its representative tag mining will facilitate the discovery of the candidate genes with discriminating gene expression. AVAILABILITY: This software and user manual are freely available at ftp://sage@bio.kuas.edu.tw/Extract-SAGE.zip. PMID- 19293994 TI - ScanMoment: a web server for combinatorial analysis of basic residues in nucleic acid binding sites. AB - ScanMoment is a webserver designed to identify the presence of the basic faced alpha-helix (BFAH) motif in the nucleic acid binding sites of proteins. The program calculates the 'Basic Moment', a parameter that quantitizes the distribution of basic residues on the surface of an alpha-helix. A sliding window is used to generate a plot displaying regions of the protein sequence that possesses a high Basic Moment and hus likely to possess a BFAH motif. The user may vary the periodicity from that of an alpha-helix (100 degrees ), to those of other secondary structures such as beta sheets and 3(10) helices. The program can also plot the periodicity of basic residues in a protein sequence using a Fourier transformation. The procedure has been used to characterize the presence of BFAHs in the N-terminal extensions of the eukaryotic aminoacyl-tRNA synthetases and to indicate the presence of a BFAH in the tRNA binding site of alanyl-tRNA synthetase. AVAILABILITY: www.scanmoment.org. PMID- 19293995 TI - RICHEST--a web server for richness estimation in biological data. AB - Richness is defined as the number of distinct species or classes in a sample or population. Although richness estimation is an important practice, it requires mathematical and computational methods that are challenging to understand and implement. We have developed a web server, RICHness ESTimator (RICHEST), which implements three non-parametric statistical methods for richness estimation. Its user-friendly web interface allows users to analyze and compare their data conveniently over the web. AVAILABILITY: A web server hosting RICHEST is accessible at http://richest.cgb.indiana.edu/cgi-bin/index.cgi and the software is freely available for local installations. PMID- 19293996 TI - CHPVDB--a sequence annotation database for Chandipura virus. AB - Databases containing proteomic information have become indispensable for virology studies. As the gap between the amount of sequence information and functional characterization widens, increasing efforts are being directed to the development of databases. For virologist, it is therefore desirable to have a single data collection point which integrates research related data from different domains. CHPVDB is our effort to provide virologist such a one-step information center. We describe herein the creation of CHPVDB, a new database that integrates information of different proteins in to a single resource. For basic curation of protein information, the database relies on features from other selected databases, servers and published reports. This database facilitates significant relationship between molecular analysis, cleavage sites, possible protein functional families assigned to different proteins of Chandipura virus (CHPV) by SVMProt and related tools. AVAILABILITY: The database is freely available at http://chpvdb.biomedinformri.org/. PMID- 19293997 TI - Docking study of triphenylphosphonium cations as estrogen receptor alpha modulators. AB - Virtual high throughput screening (VHTS) was performed to assess possible interactions which might occur between commercially available triphenylphosphonium (TPP) cations and estrogen receptor alpha (ERalpha) that could be exploited to design novel ERalpha modulators. One application of TPP cations is for delivering bioactive molecules to targets in mitochondria as the large membrane potential of mitochondria leads cations to accumulate inside them. The estrogen receptors (ERs) alpha and beta, normally activated by the endogenous hormone 17beta-estradiol, are responsible for controlling transcription of nuclear DNA necessary for human development and reproduction. ERs are also associated with the plasma membrane and have been found in the mitochondria of a variety of cell types. Selective estrogen receptor modulators (SERMs) are synthetic compounds which are used to modulate ER activity. Different SERMs display varying combinations of agonistic, antagonistic and neutral effects upon estrogen receptors depending upon the tissue type and cellular location of the receptor. Thus, they are being employed to treat a range of ER-related disorders. A common feature shared by many SERMs is the close arrangement of three aromatic rings similar to TPP cations. Given this structural similarity, the estrogenic activity of triphenyl phosphonium salts was investigated using the automated docking program eHiTS. Compounds were docked into ten different crystal structures of ERalpha. Structures were chosen based upon eHiTS ability to accurately identify the majority of estrogenically active compounds given a set of active and decoy molecules. The results of the VHTS suggest hybrids of TPP cations and known SERMs could serve as potent mitochondrial SERMs. PMID- 19293998 TI - Insights from Streptococcus pneumoniae glucose kinase structural model. AB - Streptococcus pneumonia is the common cause of sepsis and meningitis. Emergence of multiple antibiotic resistant strains in the community-acquired bacterium is catastrophic. Glucose kinase (GLK) is a regulatory enzyme capable of adding phosphate group to glucose in the first step of streptomycin biosynthesis. The activity of glucose kinase was regulated by the Carbon Catabolite Repression (CCR) system. Therefore, it is important to establish the structure-function relation of GLK in S. pneumoniae. However, a solved structure for S. pneumoniae GLK is not available at the protein data bank (PDB). Therefore, we created a model of GLK from S. pnemoniae using the X-ray structure of Glk from E. faecalis as template with MODELLER (a comparative modeling program). The model was validated using protein structure checking tools such as PROCHECK, WHAT IF and ProSA for reliability. The active site amino acid Asp114 in the template is retained in S. pneumoniae GLK model (Asp115). Solvent accessible surface area (ASA) analysis of the GLK model showed that known key residues playing important role in active site for ligand binding and metal ion binding are buried and hence not accessible to solvent. The information thus discussed provides insight to the molecular understanding of glucose kinase in S. pneumoniae. PMID- 19293999 TI - Evaluation of optimization techniques for variable selection in logistic regression applied to diagnosis of myocardial infarction. AB - Logistic regression is often used to help make medical decisions with binary outcomes. Here we evaluate the use of several methods for selection of variables in logistic regression. We use a large dataset to predict the diagnosis of myocardial infarction in patients reporting to an emergency room with chest pain. Our results indicate that some of the examined methods are well suited for variable selection in logistic regression and that our model, and our myocardial infarction risk calculator, can be an additional tool to aid physicians in myocardial infarction diagnosis. PMID- 19294000 TI - Use of sequence motifs as barcodes and secondary structures of internal transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae). AB - Most phylogenetic studies using current methods have focused on primary DNA sequence information. However, RNA secondary structures are particularly useful in systematics because they include characteristics that give "morphological" information which is not found in the primary sequence. Also DNA sequence motifs from the internal transcribed spacer (ITS) of the nuclear rRNA repeat are useful for identification of trematodes. The species of liver flukes of the genus Fasciola (Platyhelminthes: Digenea: Fasciolidae) are obligate parasitic trematodes residing in the large biliary ducts of herbivorous mammals. While Fasciola hepatica has a cosmopolitan distribution, the other major species, i.e., F. gigantica is reportedly prevalent in the tropical and subtropical regions of Africa and Asia. To determine the Fasciola sp. of Assam (India) origin based on rDNA molecular data, ribosomal ITS2 region was sequenced (EF027103) and analysed. NCBI databases were used for sequence homology analysis and the phylogenetic trees were constructed based upon the ITS2 using MEGA and a Bayesian analysis of the combined data. The latter approach allowed us to include both primary sequence and RNA molecular morphometrics and revealed a close relationship with isolates of F. gigantica from China, Indonesia and Japan, the isolate from China with significant bootstrap values being the closest. ITS2 sequence motifs allowed an accurate in silico distinction of liver flukes. The data indicate that ITS2 motifs (50%). Multivariate logistic regression analyses found the following associations: ERBB2 tumors with apocrine differentiation (p=0.0031); Triple negative tumors with high Ki-67 index (p<0.0001) and CK5 positivity (p<0.0001); HER2 negative-low receptor positive tumors (luminal B) with increased lymph node involvement (p=0.0141). The immunohistologic criteria were validated on a different set of 359 cases treated with neoadjuvant chemotherapy, which showed a pathologic complete response predominantly in ERBB2 and triple negative tumors. Immunohistochemistry is a reliable surrogate tool to classify breast carcinoma according to the gene expression profile classification. PMID- 19294004 TI - Absence of human herpesvirus 8 in pemphigus and bullous pemphigoid. AB - Pemphigus and pemphigoid are vesicobullous disorders characterized by an autoimmune attack on intercellular or basement membrane antigens, resulting in defective keratinocyte adhesion. Recently there have been reports of human herpesvirus 8 (HHV8) associated with cases of pemphigus using polymerase chain reaction (PCR) techniques, in situ hybridization, and serologic data. However, data to date is contradictory, and the relationship between this virus and autoimmune vesiculobullous disorders is unclear. No reports have attempted immunohistochemical localization of HHV8 in tissue affected by PV or BP. We studied immunohistochemical expression of HHV8 on paraffin-embedded tissue in 10 cases of pemphigus vulgaris (PV), 1 case of pemphigus foliaceous (PF) and 14 cases of bullous pemphigoid (BP). Five cases of normal skin were included as controls. Confirmatory PCR for HHV8 was performed on 4 selected cases, including 2 cases of PV and 2 cases of BP. Immunohistochemistry failed to identified the presence of HHV8 in all cases of PV (10 cases), PF (1 case) and BP (14 cases). Molecular detection of HHV8 DNA was not detected in selected PV (2 cases) and BP (2 cases). Published studies have shown contradictory evidence regarding the presence of HHV8 in vesiculobullous diseases such as pemphigus and pemphigoid. Our results refute a causal relationship between HHV8 and PV, PF and BP. PMID- 19294005 TI - Estrogen regulates vesicle trafficking gene expression in EFF-3, EFM-19 and MCF-7 breast cancer cells. AB - Estrogens are critical mediators of breast tumorigenesis. This occurs via the action of estrogens on the estrogen receptor (ER), which regulates the transcriptome of breast cancer cells. Despite the long history of the search for estrogen-regulated genes in breast cancer, knowledge of the E2-regulated transcriptome and its effects is incomplete. We used Affymetrix GeneChips to profile the effects of estradiol on the expression of genes in EFF-3, EFM-19 and MCF-7 cells. In addition to many well-characterized estrogen-regulated genes, this identified a novel group of genes that have roles in vesicle trafficking, including exocytosis. Recent evidence in the literature supports a role for vesicle trafficking in tumorigenesis. We focused on five genes (SYTL5, RAB27B, SNX24, GALNT4 and SLC12A2/NKCC1/BSC2) and confirmed their estrogen-regulation using quantitative real-time PCR (qPCR). qPCR also demonstrated that these five genes were expressed in invasive breast carcinoma tissue. Immunohistochemistry showed expression of SYTL5 in cells of normal breast ductal epithelium, ductal carcinoma in-situ (DCIS) and invasive breast carcinoma. The results suggest that a significant effect of estrogens is to regulate the expression of genes that affect diverse aspects of vesicle trafficking including exocytosis. PMID- 19294006 TI - Immunohistochemical detection of estrogen and progesterone receptor and HER2 expression in breast carcinomas: comparison of cell block and tissue block preparations. AB - Fine needle aspiration (FNA) is a rapid tool for detection of breast carcinomas. Evaluation of estrogen and progesterone receptors (ER, PR) and HER2 expression by immunohistochemistry (IHC) are routinely performed in breast carcinomas. Formalin fixation of tissue for a minimum of 6 hours, and for HER2 not more than 48 hours is the current recommended practice. In this retrospective study, we compared ER, PR and HER2 expression in breast carcinomas using archival ethanol-fixed FNA cell block with formalin fixed resection tissue block preparations. 34 archival breast carcinoma FNA cell blocks of primary origin with subsequent resection tissue blocks were identified retrospectively. All 34 cases were diagnosed as invasive ductal carcinoma. Cases with neoadjuvant or adjuvant chemotherapy were excluded. Cell blocks were initially fixed in 50% ethanol (4-12 hrs), followed by formalin fixation (minimum 6 hrs). Tissue blocks were formalin-fixed within 4-8 hrs for 6 48 hrs. ER, PR, and HER2 IHC results on the cell blocks and tissue blocks were compared. Alcohol fixed cell block samples for detection of ER and PR by IHC show good agreement with tissue block samples and are therefore a reliable method (weighted Kappa of 0.773 and 0.785, respectively) to triage patients for hormonal treatment. However, HER2 results show only moderate agreement with a weighted kappa of 0.571. The increase in discrepant results may be due to ethanol fixation which results in false positive increased HER2 expression. These results demonstrate the importance of adherence to the College of American Pathologists/ASCO guidelines for HER2 IHC. PMID- 19294007 TI - The columnar-lined mucosa at the gastroesophageal junction in non-human primates. AB - Despite that anatomists consider the cardia as a portion of the stomach, there is disagreement in the literature over whether the cardia mucosa, described as columnar-lined with mucus-producing glands (CLMMG) with or without occasional interspersed oxyntic cells, is part of the stomach, part of the esophagus or a distinct entity. For some authors this mucosa phenotype is a metaplastic glandular change of the distal esophagus caused by protracted gastro-esophageal reflux (GER). In this survey, the presence of CLMMG mucosa was searched for at the esophagus-gastric junction in 50 non-human primates (NHP). The length of the CLMMG (between the squamous epithelium of the esophagus and the first oxyntic fundic gastric gland) was assessed by the aid of an ocular microscale. In all three foetuses, all four stillborn baboons and one 4 day old baboon, the columnar lined mucosa showed depressions that corresponded to early epithelial pits without glands. In the remaining 45 post-natal NHP, the length of the CLMMG mucosa varied from 0.8 mm to 25.2 mm, and the CLMMG mucosa had replaced the distal esophageal squamous epithelium. The size was neither influenced by the post-natal age nor by the gender of the animals. In NHP, regurgitation with rumination is a natural physiological process leading to GER. The present investigation substantiates the notion that the columnar-lined mucosa with mucus producing glands is a post-natal developmental process in NHP. These animals seem to offer an excellent spontaneous model to study the series of histological events that take place in the distal esophagus of NHP, most likely under the influence of protracted GER. PMID- 19294008 TI - Clinicopathologic features of renal cell carcinoma in young adults: a comparison study with renal cell carcinoma in older patients. AB - To evaluate the clinicopathologic features of renal cell carcinoma in younger adults (40 years). The tumor size of the young adult group were smaller (5.3 vs 5.9 cm) and presented at less advanced stages (T3/T4 tumors, 18% vs 31%) than those occurring in the older age group (>40 years of age). The incidences of chromophobe RCC (12% vs. 6%) and of collecting duct carcinoma (5% vs 0.5%) were higher in the young adult group. The rate of nodal or distant metastasis was lower in young adult group (5% vs. 8.3%). More patients underwent partial nephrectomy in younger than older age group (30% vs 19%). There was no overall survival difference at 5 years (77% vs 70%), but there was a trend for a favorable survival in young adults at 10 years (77% vs 52%). In conclusion, RCC are relatively infrequent in patients who are younger than 40 years. The tumors in this group appear to be smaller and less advanced at presentation. Chromophobe RCC and collecting duct carcinoma are more frequently seen. More patients undergo partial nephrectomy and overall long term survival appears to be more favorable. PMID- 19294009 TI - Disseminated coccidioidomycosis: a case report from the United kingdom. AB - We reported the first case of disseminated coccidioidomycosis from the UK successfully treated with amphotericin B, where skin biopsy was the initial clue for the correct diagnosis. PMID- 19294010 TI - Fatal cerebral hemorrhage in a patient with CD19-positive IgM-producing aggressive plasma cell myeloma, hyperviscosity syndrome and cryoglobulinemia. AB - IgM plasma cell myeloma (PCM) is a rare entity, and CD19 positivity is found in only 1-4% of PCM. Here we report a unique case of IgM PCM, in which the plasma cells are positive for CD19. Clinically, the patient presented with hyperviscosity syndrome, mimicking the clinical manifestation of Waldenstrom's macroglobulinemia. In addition, the IgM para-protein from the patient behaved like cryoglobulins, which interfered with some of the laboratory measurements and resulted in erroneous platelet count, mean platelet volume, and serum IgM level. Despite chemotherapy, the PCM persisted and progressed to plasma cell leukemia, and the patient died of a left frontal hematoma with widespread cerebral hemorrhage extending from left lateral ventricle, third ventricle, fourth ventricle, to cisterna magna. This case represents the first CD19+ IgM-producing PCM and the second case of brain hemorrhage due to plasma cell myeloma/leukemia. PMID- 19294011 TI - Stimulation of human CD4+ T lymphocytes via TLR3, TLR5 and TLR7/8 up-regulates expression of costimulatory and modulates proliferation. AB - The cells of innate and adaptive immunity, although activated by different ligands, engage in cross talk to ensure a successful immune outcome. Toll-like receptors (TLRs) are key components of the innate immune system and have the ability to detect microbial infection and trigger host defence responses. Otherwise, human T lymphocytes are able to produce most TLRs. Thus, we analyze the capability of some TLR ligands to modulate the function of highly-purified CD4+ T cells. We found that agents acting via TLRs (poly I:C, a TLR3 ligand; flagellin, a TLR5 ligand; and R848, a TLR7/8 ligand) are able to regulate the expression of costimulatory molecules both on purified antigen presenting cells and on purified T lymphocytes. Moreover, the activation mediated by TLRs determines a kinetic expression of B7-family members such as through an inhibition of T lymphocytes delayed proliferation. These findings suggest a functional role of some invading microorganisms in regulating acquired immunity. PMID- 19294012 TI - Utilisation of carbon sources by pythium, phytophthora and fusarium species as determined by biolog(r) microplate assay. AB - This study examined the metabolic activity of pure cultures of five root pathogens commonly found in closed hydroponic cultivation systems (Phytophthora cryptogea (PC), Phytophthora capsici (PCP), Pythium aphanidermatum (PA), Fusariumoxysporum f.sp. radicis-lycopersici (FORL) and Fusarium solani (FS)) using sole carbon source utilisation in order to develop effective biocontrol strategies against these pathogens. Aliquots of 150 microL of the mycelial suspension were inoculated in each well of GN2 microtitre plates. On the basis of average well colour development and number of positive wells, the pathogens were divided into two groups, (i) PA and FORL and (ii) PC, PCP and FS. Group (i) was characterised by a short lag-phase, a rapid exponential phase involving almost all carbon sources offered and a long stationary phase, while group (ii) had a more extended lag-phase and a slower utilisation rate of the carbon sources offered. The three isolates in group (ii) differed significantly during their exponential phase. The lowest utilisation rate of carbon sources and number of sources utilised was found for PCP. Of the major group of carbon sources, six carbohydrates, three carboxylic acids and four amino acids were rapidly used by all isolates tested at an early stage. The carbon sources gentibiose, alpha-D glucose, maltose, sucrose, D-trehalose, L-aspartic acid, L-glutamic acid, L proline persisted to the end of the exponential phase.Moreover, similarities between the metabolic profiles of the tested pathogen and the those of the resident microflora could also be found. These findings are of great importance as regards the role of the resident microflora in the biocontrol. PMID- 19294013 TI - Heterodera raskii n. sp. (Heteroderidae: Tylenchina), a Cyst Nematode on Grass, from Hyderabad, India. AB - Heterodera raskii n. sp. is described and illustrated from specimens collected from roots of bulb grass, Cyperus bulbosus, in Hyderabad, India. The new species belongs to the 'goettingiana' group and differs from closely related H. cyperi by the elongate ovoid shaped cysts and females, greater fenestral length, width, vulval slit, and absence of egg sac. The stylet knob shape was round in second stage juveniles and posteriorly sloping in females and males of H. raskii n. sp., while it was anteriorly directed in second-stage juveniles and spherical in females and males of H. cyperi. PMID- 19294014 TI - Ecology and Control of Cereal Cyst Nematode (Heterodera avenae) in Southern Australia. AB - The ecology and control of cereal cyst nematode in southern Australia is reviewed. The wide distribution of Heterodera avenae in Victoria and South Australia is due largely to movement of cysts by wind during dust storms. The fungus Rhizoctonia solani frequently is associated with the nematode in a disease complex in wheat, and disease symptoms are most severe on lighter or well structured soils. Crop rotations which include periods of fallow, or of nonhost crop reduce population levels of H. avenae and improve yields. Early-sown crops (April-May) are less severely damaged than late-sown crops (June-July). The resowing of damaged wheat crops or the application of nitrogenous fertilizers rarely improve grain yields. 'Katyil,' the world's first wheat cultivar bred specifically with resistance to H. avenae, has been released in Victoria. Chemical control of the nematode in cereals is now commercially feasible, and five nematicides are registered for use by growers. PMID- 19294015 TI - Morphological and Histochemical Changes Occurring during the Life-span of Root tip Galls on Lolium perenne Induced by Longidorus elongatus. AB - The RNA and protein content of perennial ryegrass root-tip galls induced by Longidorus elongatus were measured from transverse sections and the morphology described. Galls progressed through five distinct stages and were viable for only 10-12 days at 18 C, after which they collapsed and became necrotic. In the initial stage hypertrophy occurred and cells contained enlarged nuclei and nucleoli, a greater proportion of cytoplasm, and increased concentrations of protein. This was followed by hyperplasia; cells divided to give two or four daughter cells, accompanied by a proportionate reduction in volumes of cytoplasm, nuclei, and nucleoli and reduced concentrations of RNA and protein. The third stage was secondary hypertrophy with enlarged, amoeboid nuclei and nucleoli and a significant increase in concentration of RNA and protein. In the final two stages, as feeding by L. elongatus progressively removed cell contents, most cells were devoid of inclusions and galls collapsed and were invaded by soil bacteria. This ordered development and exploitation of galls suggests that L. elongatus may have two phases in its feeding. PMID- 19294016 TI - Joint Influence of Pratylenchus penetrans (Nematoda) and Leptinotarsa decemlineata (Insecta) on Solanum tuberosum Productivity and Pest Population Dynamics. AB - The joint action of a plant parasitic nematode, Pratylenchus penetrans (root lesion nematode), and an insect defoliator, Leptinotarsa decemlineata (Colorado potato beetle), on growth, development, and yield of Solanum tuberosum cv. Superior was studied in the field. Three population densities of P. penetrans were superimposed on each of three population levels of L. decemlineata. The major impact of P. penetrans on final yield was through a reduction in the number of tubers formed during tuber initiation. Defoliation by L. decemlineata increased with time as larvae advanced through successive instars and densities increased. This resulted in a significant reduction in tuber weight and numbers. Total yield of S. tuberosum was decreased by 66% with increasing population densities of L. decemlineata and 27 % with increasing densities of P. penetrans. L. decemlineata feeding did not affect soil population densities of P. penetrans. Root population densities of P. penetrans, however, were significantly (P = 0.05) higher in plants maintained beetle free than in plants grown in the presence of the beetles. PMID- 19294017 TI - Rhyssocolpus arcticus n. sp. (Nematoda: Nordiidae) from Ellesmere Island and a Redescription of Lindseyus costatus Ferris and Ferris, 1973 (Nematoda: Roqueidae) from Quebec, Canada. AB - A new species of Rhyssocolpus from Ellesmere Island in the Canadian arctic is described and illustrated. Its distinctive features are the set off head and the extensively striated cuticle in the vulval area with well-developed vulval flaps. The male has small spindle-shaped sperm 4-5 mum in length, and the adanal supplement is separated from the anterior series of 7-11 supplements by a distinct gap. A key to the species of Rhyssocolpus is provided. Lindseyus costatus from the St. Lawrence River, Quebec, is also redescribed. Notable differences from the original description are the irregular muscle bands of the sinistrally spiraled esophageal sheath, smooth exocuticle, longer prerectum, shorter male tail, and longer spicules. New information on the esophageal gland nuclei and sperm size and shape is also provided. PMID- 19294018 TI - Population Development and Pathogenicity of Meloidogyne javanica on Flue-cured Tobacco as Influenced by Ethoprop and DD. AB - Growth of flue-cured tobacco as influenced by Meloidogyne javanica and the effectiveness of DD and ethoprop to manage this nematode were evaluated over two growing seasons. Populations of M. javanica, root galling, plant height, steam crown diameter, whole plant weight, and yield were monitored at approximately 2 week intervals beginning 28 days after transplanting. Treatment influence on nematode population development, root galling, and plant growth generally followed a pattern in descending order of efficacy: DD (187 liters/ha), ethoprop (27, 18, or 9 kg a.i./ha), and control. In all treatments, nearly season-long increases in M. javanica populations and root galling were observed. Correlation coefficients relating nematode populations or root galling to final tobacco yield suggested either method may be used successfully to evaluate nematicide efficacy in research plots. Plant growth parameters most affected by M. javanica in order of decreasing severity were cured leaf yield, whole plant weight, plant height, and stem diameter. PMID- 19294019 TI - Probability range in damage predictions as related to sampling decisions. AB - The risk involved in basing a nematode management decision on predicted crop loss is related to the uncertainty in the crop damage function and error in measuring nematode population density. The sampling intensity necessary to measure a nematode population with specified precision varies with population density. Since the density is unknown prior to sampling, optimum sampling intensity for a management decision is calculated for the economic threshold population level associated with the management cost. Population densities below the threshold are measured with greater precision than required; those above the threshold are less precisely measured, but invoke management. The approach described provides resolution to sampling strategies and allows assessment of the risk associated with the management decision. PMID- 19294020 TI - Gastromermis kolleonis n. sp. (Nematoda: Mermithidae), a Parasite of Midges (Chironomus sp. Chironomidae) from Argentina. AB - Gastromermis kolleonis n. sp. (Nematoda: Mermithidae) is described from the Arroyo Saldan River in Cordoba, Argentina. This species parasitizes midges of the genus Chironomus (Chironomidae: Diptera). It is distinguished from other members of the genus by the presence of six longitudinal chords, vulval flaps, degree of ventral displacement of the mouth, and size and shape of the spicule and amphids. PMID- 19294021 TI - Observations on the Mode of Parasitism and Histopathology of Meloidodera floridensis and Verutus volvingentis (Heteroderidae). AB - Some aspects of the host-parasite relationships of two heteroderid nematodes are described. Meloidodera floridensis induced formation of single uninucleate giant cells in the stelar parenchyma tissue of sand pine (Pinus clausa) roots. Wrinkling and yellowing of the cuticle were associated with maturation of the adult female (cystoid stage). The mode of parasitism of different life stages of Verutus volvingentis on buttonweed (Diodia virginiana) is described. The nematode caused extensive necrosis during penetration and the formation of a large feeding site consisting of nonhypertrophied parenchyma cells with enlarged nuclei and thickened cell walls in the cortex. Walls between cells within the feeding site degenerated, resulting in the formation of a syncytium. Two citrus rootstocks, rough lemon (Citrus lirnon) and trifoliate orange (Poncirus trifoliata), were not hosts of V. volvingentis. PMID- 19294022 TI - Description of Meloidoderita polygoni n. sp. (Nematoda: Meloidoderitidae) from USA and Observations on M. kirjanovae from Israel and USSR. AB - Meloidoderita polygoni n. sp. is described and illustrated from roots of smartweed (Polygonum hydropiperoides) from Beltsville, Maryland. This new species is similar to M. kirjanovae but differs especially in having larger spines on the cystoid bodies, females with the anus much closer to the vulva, and more posterior excretory pore. M. polygoni differs from M. safrica particularly in having females with a shorter stylet, a DGO much closer to base of stylet, greater distance between vulva and anus, and larger cystoid bodies. LM and SEM observations showed only three incisures in lateral fields of juveniles and males and no bursa in males. Morphometric data and illustrations are given for M. kirjanovae from mint (Mentha longifolia) in Israel and some details on a limited number of specimens from Armenian SSR. LM examination of juveniles from both these areas indicated only three incisures in lateral fields. Males from Israel had no detectable bursa and appeared to have only three incisures in lateral fields. (Males from Armenian SSR not observed.). PMID- 19294023 TI - Anhydrobiosis in Pratylenchus penetrans. AB - Anhydrobiotic survival of Pratylenchus penetrans was compared in several soil moisture regimes. Bodies of anhydrobiotic nematodes were coiled. In slow-dried soils, Vineland silt loam (VSL) and Fox loamy sand (FLS), 70 and 58% of the total P. penetrans populations were anhydrobiotic when soil moistures reached ca. 3% and water potential 15 kPa or greater. Coiling began at a much lower water potential in FLS than in VSL. In fast-dried soils, only 31 and 22% of the P. penetrans populations in the same two soil types had entered the anhydrobiotic state at comparable moistures. In the above soils, 76-96% of the P. penetrans were alive immediately after entering the anhydrobiotic state. In slow-dried VSL, some nematodes (1%) survived 770 days. In the other soils, all anhydrobiotic nematodes were dead after 438 days. Anhydrobiosis increased the ability of nematodes to survive subzero temperatures, but it did not increase their ability to survive temperatures above 40 C. Infectivity and reproductivity of rehydrated P. penetrans were not affected by anhydrobiosis. PMID- 19294024 TI - Tolerance to Heterodera glycines in Soybean. AB - Fifty-four susceptible soybean, Glycine max, cultivars or plant introductions were evaluated for tolerance to H. glycines, the soybean cyst nematode (SCN). Seed yields of genotypes were compared in nematicide-treated (1,2-dibromo-3 chloropropane, 58 kg a.i./ha) and nontreated plots at two SCN-infested locations over 3 years. Distinct and consistent levels of tolerance to SCN were observed among soybean genotypes. PI 97100, an introduction from Korea, exhibited the highest level of tolerance with an average tolerance index ([yield in nontreated plot / yield in nematicide-treated plot] x 100) of 96 over 2 years. Coker 156 and Wright had moderate levels of tolerance (range in index values 68 to 95) compared to the intolerant cuhivars Bragg and Coker 237 (range in index values 33 to 68). Most of the soybean genotypes evaluated were intolerant to SCN. The rankings of five genotypes for tolerance to SCN and Hoplolaimus columbus were similar. Tolerance for seed yield was more consistently correlated with tolerance for plant height (r = 0.55 to 0.64) than for seed weight (r = 0.23 to 0.65) among genotypes. PMID- 19294025 TI - Effect of Phytotoxin from Nematode-induced Pinewilt on Bursaphelenchus xylophilus and Ceratocystis ips. AB - A phytotoxic extract from Pinus sylvestris infected with Bursaphelenchus xylophilus inhibited growth of the blue-strain fungus Ceratocystis ips and caused temporary paralysis in vitro of B. xylophilus. Although the nematodes recovered from paralysis, final population size of B. xylophilus was suppressed by the toxin. Extracts from noninfected P. sylvestris affected neither the fungus nor the nematode. PMID- 19294026 TI - Changes in the Reproduction of Heterodera glycines on Different Lines of Glycine max. AB - Selection for ability of soybean cyst nematode (SCN), Heterodera glycines, to reproduce on soybeans with different sources of resistance divides some SCN race 4 field populations into two distinct subpopulations. These subpopulations reproduce well on 'Bedford' and plant introduction (PI) 88788 or PI 89772 and PI 90763 but not on both pairs of soybean lines. The ability of these subpopulations to reproduce on the four soybean lines was reversed by changing the soybean line used as a host during a second cycle of selection. When SCN populations previously selected for reproduction on Bedford and PI 88788 were selected for their ability to reproduce on D72-8927 and J74-88, the ability of these populations to reproduce on Bedford and PI 88788 decreased significantly and their ability to reproduce on PI 89772 and PI 90763 increased significantly. Conversely, when SCN populations, previously selected for reproduction on P189772 and P190763, were selected for their ability to reproduce on Bedford, the reproduction of these populations on Bedford increased significantly and reproduction on PI 89772 and PI 90763 decreased significantly. Selection for ability of a SCN race 4 field population to reproduce on soybean lines derived from SCN race 4 resistant PIs resulted in the same division of the field population into two distinct subpopulations. These data substantiate earlier proposals to rotate cultivars with different genes for SCN resistance as a means of managing SCN populations. PMID- 19294027 TI - Effect of Soil Temperature on Reproduction of Meloidogyne chitwoodi and M. hapla Alone and in Combination on Potato and M. chitwoodi on Rotation Plants. AB - Meloidogyne chitwoodi developed and reproduced more rapidly than M. hapla in potato roots at 15, 20, or 25 C when both species of nematodes were inoculated simultaneously at 250 or 1,000 juveniles of each. At 30 C significantly more M. hapla than M. chitwoodi females were found at the lower inoculum level after 41 days. More M. chitwoodi than M. hapla juveniles were extracted from soil at 15, 20, and 25 C, but only at the lower inoculum level at 30 C. Potato was considered a more suitable host for M. chitwoodi than M. hapla because of M. chitwoodi's greater reproduction at 15, 20, and 25 C. Corn and wheat cultivars tested supported M. chitwoodi reproduction at temperatures of 10, 15, 20, and 25 C, but fewest eggs were produced on these plants at 20 C. Temperatures of 10 to 25 C had little influence on the low reproduction of M. chitwoodi on four alfalfa cultivars. M. chitwoodi reproduced on the alfalfa entry Mn PL9HF. PMID- 19294028 TI - Distribution of Tylenchulus semipenetrans in a Texas Grapefruit Orchard. AB - Distribution of the citrus nematode (Tylenchulus semipenetrans) was studied over 18 months in a 6-year-old orchard of grapefruit (Citrus paradisi cv. Ruby Red) on sour orange (C. aurantium) rootstock. The 1.8-ha orchard was under chemical weed control, no tillage, and flood irrigation. Highest numbers of nematodes were found in the top 15 cm of the soil profile. The nematode population peaked in April and declined to lowest levels in August and September. Numbers of nematodes were negatively correlated (r = -0.95) with soil temperatures above 29 C. Soil populations of nematodes were not correlated with soil moisture. The distribution of the nematode in the field was highly skewed and was described by a negative binomial. In this 1.8-ha block, five soil samples of 12 cores each would provide an estimate within 20% of the true nematode population mean with 95% confidence. PMID- 19294029 TI - Response of Peach Seedlings to Infection by the Root Lesion Nematode Pratylenchus penetrans under Controlled Conditions. AB - Twenty-one open pollinated populations of peach rootstock seedlings were evaluated for their response to infection by the root lesion nematode, Pratylenchus penetrans, over a period of 98 days. Nematode-infected peach seedling populations were shorter in plant height and had less shoot weight but more dry root weight than nematode-free controls. Rootstock differences were demonstrated for nematode increase over the 98-day period, and average total numbers of nematodes in soil and roots. Rootstocks were classified into three groups differing in total nematode population levels, ratio of nematode increase, and the number of nematodes per root. The heritable nature of rootstock response to nematodes was evident. Rootstocks showing the lowest response to nematode infection included Tzim Pee Tao, Rutgers Red Leaf, and two progenies of a cross of these two rootstocks. PMID- 19294030 TI - Crop Rotation and Races of Meloidogyne incognita in Cotton Root-knot Management. AB - The influence o f various crop rotations and nematode inoculum levels on subsequent population densities of Meloidogyne incognita races 1 and 3 were studied in microplots. Ten different 3-year sequences o f cotton, corn, peanut, or soybean, all with cotton as the 3rd-year crop, were grown in microplots infested with each race. Cotton monoculture, two seasons o f corn, or cotton followed by corn resulted in high race 3 population densities and severe root galling on cotton the 3rd year. Peanut for 2 years preceding cotton most effectively decreased the race 3 population and root galls on cotton the 3rd year. Race 1 did not significantly influence cotton growth or yield at initial populations of up to 5,000 eggs/500 cm(3) soil. At 5,000 eggs/500 cm(3), cotton growth was suppressed by race 3 but yield was not affected. PMID- 19294031 TI - Nematostatic Activity of Oxamyl and N,N-Dimethyl-1-cyanoformamide (DMCF) on Meloidogyne incognita Juveniles. AB - The nematostatic activity of oxamyl, methyl-N',N'-dimethy]-N-hydroxy-l thiooxamimidate (oxamyl-oxime) and N,N-dimethyl-l-cyanoformamide (DMCF) was studied by immersing 10 Meloidogyne incognita second-stage juveniles into aqueous solutions of various concentrations of each chemical. At concentrations of 500 to 8,000 mug/ml, oxamyl quickly immobilized immersed juveniles. In all other concentrations studied (down to 4 mug/ml), oxamyl stopped or reduced movement of juveniles within 24 hours. DMCF also quickly immobilized juveniles at concentrations of 4,000 and 8,000 mug/ml and reduced movement at 2,000 mug/ml. Lower concentrations had no observed effect on movement. In solutions of the oxime from 2,000 to 8,000 mug/ml, some reduction of movement was observed, but most juveniles maintained some motion over a period of 24 hours. Juveniles were transferred to water from 4,000 mug/ml solutions of oxamyl and DMCF after various intervals of time in order to determine the effect of duration of exposure to the chemicals on the ability of the immobilized juveniles to recover normal motion. Some recovery was observed even after 24 hours of exposure to DMCF, but none after exposure to oxamyl for longer than 40 minutes. PMID- 19294032 TI - Comparative Electrophoretic Analyses of Soluble Proteins from Heterodera glycines Races 1-4 and Three Other Heterodera Species. AB - Modified polyacrylamide gel and SDS-polyacrylamide gel electrophoretic systems using a low molarity tris-HCl buffer and equal pH of homogenizing buffer and stacking gel provided improved stacking for separation of soluble proteins from Heterodera schachtii, H. trifolii, H. lespedezae, and H. glycines races 1, 2, 3, and 4, compared with previous studies with cyst nematodes, The four Heterodera species were easily distinguished using the polyacrylamide gel system, but H. trifolii and H. lespedezae had similar protein patterns. H. glycines races were not separable by that system. The SDS-polyacrylamide gel system produced different protein patterns for all four Heterodera species although H. trifolii and H. lespedezae differed by only a single band, suggesting that these two may be subspecifically related. A protein band unique to H. glycines races 3 and 4 was not detected in SDS-polyacrylamide gel profiles from races 1 and 2. Molecular weight determinations were 55,000 for distinctive proteins in profiles of H. trifolii and 75,000 for H. glycines races 3 and 4. PMID- 19294033 TI - Preparing nematode permanent mounts with adhesive tapes. PMID- 19294034 TI - Comparison of five methods for measuring nematode volume. PMID- 19294035 TI - Technique for axenizing nematodes. PMID- 19294036 TI - Pathogenicity and Reproduction of Hoplolaimus columbus and Meloidogyne incognita on 'Davis' Soybean. AB - The effects of initial populations of Hoplolaimus columbus and Meloidogyne incognita on growth and yield of Davis soybean were determined for 1980 and 1981 in microplots and H. columbus in field tests in 1981. M. incognita suppressed yield in microplots both years and H. columbus in 1980. Maximum suppression of dry pod weight by M. incognita was 45% and by H. columbus 35%. The relationship of yield vs. nematode population at planting time was described by a declining exponential model. Maximum reproductive rates for M. incognita and H. columbus were 67.0 and 4.7, respectively, and were inversely proportional to initial population level. Nematode reproductive rates, survival ability, and feeding habits suggest species specific life strategies in the ecological community. PMID- 19294037 TI - Effect of Temperature on the Development of Thelastoma bulhoesi (Oxyurata, Thelastomatida) and Other Nematodes. AB - Embryonation of Thelastoma bulhoesi was monitored at eight temperatures between 0 and 35 C. Cell division did not occur below 15 C or at 35 C. Development was most rapid at 25 and 36 C. The effect of temperature on the rate of embryological development of T. bulhoesi at different stages was measured using the temperature coefficient, Q. The developmental temperature response curve obtained for T. bulhoesi was similar to enzyme temperature response curves. Our evidence supports the thesis that nematode embryonation, as affected by temperature, varies between species and between stages of development. PMID- 19294038 TI - New and known species of pratylenchus filipjev, 1936 (nematoda: pratylenchidae) from haryana, India, with remarks on intraspecific variations. AB - Two new monosexual and one bisexual species Pratylenchus Filipjev, 1936 collected from Haryana state of India are described and illustrated. The primary distinguishing features of these species are Pratylenchus microstylus n. sp.: L = 331-458 mum, spear = 11 or 12 mum; Pratylenchus cruciferus n. sp.: L = 648-793 mum, central core of lateral fields with oblique lines, hemizonid 2-8 annules anterior to excretory pore; Pratylenchus ekrami n. sp.: spear = 11-13 mum, spermatheca oblong, post vulval uterine sac with differentiated cells, tail with 26-40 annules, males abundant. Studies on intraspecific variations of P. cruciferus, P. ekrami, and P. coffeae (Zimmermann, 1898) Goodey, 1951 revealed that spear length and value of 'V' are the least variable characters. Body length and size of post vulval uterine sac varies to varying degrees in different species. Shape of median bulb in P. ekrami, number of incisures in P. coffeae, and tail shape in P. ekrami and P. coffeae exhibit the greatest amount of intraspecific variations. P. zeae Graham, 1936 and P. thornei Sher & Allen, 1953 are the other species collected during the present studies. PMID- 19294039 TI - Variability in Reproduction of Four Races of Meloidogyne incognita on Two Cultivars of Soybean. AB - Variability in the reproduction of the four races ofMeloidogyne incognita on the soybean cuhivars Pickett 71 and Centennial was studied in growth chamber experiments. Analysis of variance in the number of eggs produced by the races 6 weeks after the plants had been inoculated with 5,000 eggs of each race revealed that the nematode race by soybean cultivar interaction was highly significant (P = 0.001). Races 1, 3, and 4 produced from about 5,000 to 15,000 eggs per root system on Pickett 71 and only from about 300 to 600 eggs per root system on Centennial. In contrast, race 2 produced about 8,000 eggs per root system on Centennial and about 1,200 eggs per root system on Pickett 71. In a second experiment, in which the plants were inoculated with 2,000 second-stage juveniles, race 1 and race 2 produced about 13,000 and 3,000 eggs per root system, respectively, on Pickett 71 and about 600 and 10,000 eggs per root system, respectively, on Centennial. The results suggest that M. incognita resistance in soybean is race-specific. PMID- 19294040 TI - Heydenius dominicus n. sp. (Nematoda: Mermithidae), a Fossil Parasite from the Dominican Republic. AB - Heydenius dominicus n. sp. is described as a new species of fossil mermithid nematode from Dominican Republic amber. The species is represented by two specimens of parasitic juveniles that left their insect host and became embedded in the resin. The nematodes are associated with an adult male limoniid (Diptera: Limoniidae) and an adult female mosquito (Diptera: Culicidae). The parasites are thought to have emerged from the mosquito host. This is the first report of a fossil mermithid from a Neotropical area. PMID- 19294041 TI - Reproductive Isolation and Taxonomic Differentiation of Romanomermis culicivorax Ross and Smith, 1976 and R. communensis Galloway and Brust, 1979. AB - The infertility of hybrid progeny of Romanomermis communensis and R. culicivorax supports their retention as distinct species. Their taxonomic separation on the basis of morphometric data and possession of a cone-shaped spicule guide is rejected. However, differences in the enzyme patterns of peptidase and phosphoglucomutase and the restriction fragment length differences in repetitive genomic DNA provide sensitive diagnostic characters that confirm the differentiation into two species. PMID- 19294042 TI - Nematode population and community dynamics in soybean-wheat cropping and tillage regimes. AB - The nematode community structures of various soybean-wheat regimes and of a single-cropped, conventionally tilled soybean regime were studied at two sites in Tennessee. Each of the 100 nematode species identified in the study was placed in one of five trophic groups, the most diverse being plant parasites (31 species), followed by Dorylaimida (26 species), bacterivores (23 species), fungivores (15 species), and predators (5 species). No significant differences in overall diversity and dominance among treatments and trophic groups were found. Densities of Heterodera glycines Ichinohe infective juveniles were significantly higher in single-cropped, conventionally tilled soybeans in July. When data were subjected to ordination analysis, it was shown that plant-parasitic nematode communities produced an aggregation of conventionally tilled, single-cropped soybean plots when compared to all double-cropped treatments. Ordination of overall nematode communities yielded similar results. PMID- 19294043 TI - A New Gall-Forming Species of Anguina Scopoli, 1777 (Nemata: Anguinidae) on Bluegrass, Poa annua L., from the Coast of California. AB - Anguina pacificae n. sp. is described and illustrated from stem galls on bluegrass, Poa annua L., from golf courses along coastal California. The females are characterized by constrictions in the anterior and posterior connections of the isthmus with the respective parts of the esophagus, the long multicellular columella, and the sharply pointed tail tip. Males are dorsally curved after death; body width is increased markedly after 13 annuli in both sexes, and the tail is conical and with an acute terminus. PMID- 19294044 TI - Influence of Nacobbus aberrans Densities on Growth of Sugarbeet and Kochia in Pots. PMID- 19294045 TI - Cropping Effects of Marigolds, Corn, and Okra on Population Levels of Meloidogyne javanica and on Carrot Yields. PMID- 19294046 TI - Pararotylenchus hopperi nomen novum. PMID- 19294047 TI - Rangeland Grasses as Hosts of Meloidogyne chitwoodi. PMID- 19294048 TI - Flowering Delay in Flue-cured Tobacco Infected with Meloidogyne Species. PMID- 19294049 TI - Expression of Resistance in Peanuts to Pratylenchus brachyurus: Impact on Screening for Resistance. PMID- 19294050 TI - Gametogenesis and the Chromosomes of Meloidogyne nataliei: Not Typical of Other Root-knot Nematodes. AB - Studies of oogenesis and spermatogenesis revealed that Meloidogyne nataliei is a diploid, amphimictic species with four (n), relatively large chromosomes, and possibly with an XX female symbol-XY male symbol mechanism of sex determination. It differs considerably from all other amphimictic, or meiotically parthenogenetic, species of Meloidogyne which have 13-18 smaller chromosomes and from Meloidogyne (Hypsoperine) spartinae which has seven. Consequently, the taxonomic position of M. nataliei needs to be re-evaluated. The chromosomes of M. nataliei and their behavior during gametogenesis resemble more closely chromosomes of the genus Heterodera than those of the genus Meloidogyne. This resemblance, however, may not imply a closer phyletic relationship of M. nataliei to heteroderid nematodes. PMID- 19294051 TI - Use of enzyme phenotypes for identification of meloidogyne species. AB - Enzyme phenotypes were obtained for 291 populations from 16 species of Meloidogyne originating from 65 countries. Soluble proteins from macerates of individual egg-laying females were separated by electrophoresis in 0.7-mm-thick polyacrylamide gels. Enzymes investigated were nonspecific esterases, malate dehydrogenase, superoxide dismutase, and glutamate-oxaloacetate transaminase. Esterases were polymorphic and most useful in identification of major species. About 94% of the populations of M. hapla, 98% of M. incognita, and 100% of M. javanica could be identified to species on the basis of esterase phenotypes alone. About 84% of the populations of M. arenaria exhibited three distinct phenotypes. Two of them were highly species specific (accuracy of identification 98-100%). The third, and least prevalent, phenotype occurred also in two other species. Another 12 less common Meloidogyne species, of which only one or a few populations of each were studied, exhibited a variety of esterase phenotypes, some of which may prove to be species specific. Superoxide dismutase phenotypes similarly were helpful in the characterization of certain species; however, the same phenotype was often observed in more than one species. The remaining two enzymes, with few exceptions, proved to be less useful for identification of Meloidogyne species. Multienzyme phenotypes represented by two or more enzymes often offered biochemical profiles more valuable for definitive characterization of Meloidogyne species than single enzymes. PMID- 19294052 TI - An Induced Resistance Effect of Hydroxyurea on Plants Infected by Meloidogyne javanica. AB - Aqueous solutions of hydroxyurea (HU) in various concentrations were applied as soil drenches to Meloidogyne javanica-infected plants. At a concentration of 15 ppm, the chemical hampered giant cell formation and the number of females on the roots was 20% of that of the control but the growth of the host plants was not affected. Additional HU applications after the one at infection did not add to the inhibitory effect. HU exerted its effect on M. javanica-infected tomato in five soil types. Soil temperature of 32 C neutralized the HU-induced resistance in much the same way that high temperature breaks the natural resistance in M. incognita-resistant tomato. This study provides further evidence of the role of HU as an induced resistance agent. PMID- 19294053 TI - Effect of tillage system and irrigation on population densities of plant nematodes in field corn. AB - Soil populations of plant-parasitic nematodes were monitored bimonthly for 18 months in irrigated and nonirrigated corn plantings using four production systems: conventional and minimum tillage with crop residue returned and minimum tillage with 60% or 90% of previous corn crop residue removed. Populations of Meloidogyne incognita, Scutellonema brachyurum, Pratylenchus scribneri, and Paratrichodorus christiei varied among the tillage, nematicide, and irrigation treatments. Meloidogyne incognita and P. christiei populations were not significantly affected by tillage method, but S. brachyurum populations were highest during April 1981 and 1982 in minimum tillage treatments where crop debris was not removed. In contrast, S. brachyurum populations were lowest during the same period in minimum tillage plots where 90% of previous crop debris had been removed or where residues were incorporated with conventional tillage. Populations of P. scribneri were lowest in minimum tillage during August 1981 and April 1982. Regardless of tillage system, corn yields in all nonirrigated plots were increased during 1982 by application of carbofuran (2.24 kg a.i./ha). No yield increases were observed following nematicide application in 1981. PMID- 19294054 TI - Dolichodorus miradvulvus n. sp. (Nematoda: Tylenchida) with a Key to Species. AB - Dolichodorus miradvulvus n. sp. from Anubias nana Engler in Florida is described and illustrated. The female is characterized by deep grooves in the cuticle on the ventral surface just anterior and posterior to the vulva, and by transversely elongate pouches anterior and posterior to the vulva. Both sexes have a constricted area of the stylet shaft just anterior to the knobs, and on the male the intersection of the lateral field and bursa appear sclerotized. PMID- 19294055 TI - Population Dynamics and Description of Ptycholaimellus hibernus n. sp. (Nematoda: Chromadoridae). AB - Ptycholaimellus hibernus n. sp. from the muddy subtidal of North Inlet Estuary, Georgetown, South Carolina, is described and a key to the genus is provided. P. hibernus differs from all other species of Ptycholaimellus by the shape of the gubernaculum. Ptycholaimellus sp. 2 Hopper 1969 is synonymized with P. ponticus. The abundance of P. hibernus, measured over a 3-year period, is greatest from January to March, coinciding with minimal annual water temperatures (10-15 C). P. hibernus abundance was significantly (negatively) correlated with water temperature and (positively) with the depth of the anoxic sediment layer. PMID- 19294056 TI - Population Densities of Meloidogyne incognita and Yield of Capsicum annuum. AB - Two microplot experiments in 1981 and 1983 provided information on the effect of different population densities of Meloidogyne incognita race 1 and yield of sweet pepper. Microplots were square concrete pipes (30 x 30 cm and 50 cm long) filled with 40 liters of soil infested with 0, 0.062, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, 32, 64, 128, 256, and 512 eggs and juveniles/cm(3) soil. Tolerance limits of 2.2 and 0.165 eggs and juveniles/cm(3) soil and minimum yields of 58% and 20% of the controls were obtained in 1981 and 1983, respectively. Maximum reproduction rates of the nematode were 274 and 1,498 at the lowest initial population density. The population of the nematode declined rapidly after harvest, and only 13% and 6.5% of eggs and juveniles were detected in the soil after 1 and 6 months, respectively. PMID- 19294057 TI - Fungal Parasites of the Cereal Cyst Nematode Heterodera avenae in Southern Sweden. AB - Fungal parasites of the cereal cyst nematode Heterodera avenae were isolated from four sites in southern Sweden. In all, 15 different fungi were isolated from different stages of the nematode life cycle. Among the egg parasites, Verticillium chlamydosporium was common in young cysts on roots, whereas an unidentified species of Verticillium (Verticillium sp. 1) was the dominating species in cysts from soil, especially if the soll had been stored for 8-12 months. V. chlamydosporium was frequently isolated from eggs in cysts from soil, when analyzed shortly after sampling. Verticillium sp. 1 is distinct from V. chlamydosporium because it does not produce dichtyo-chlamydospores in the aerial mycelium and because it grows at 6 C where V. chlamydosporium fails to grow. Paecilomyces lilacinus, Microdochium bolleyi, Cylindrocarpon sp., and several nonsporulating fungi were also isolated from eggs in cysts from soil. Between 10 and 20% of the eggs in cysts collected in the field were infected with fungi. In a pot test between < 1 and 29%, with a mean of 13%, of females on roots became infected, always by Nematophthora gynophila. Resting spores of N. gynophila extracted directly from field soil, collected at the four sites, varied from 3 to 49 spores/gram of air dried soil. PMID- 19294058 TI - Influence of Glomus fasciculatum and Meloidogyne hapla on Allium cepa in Organic Soils. AB - The influence of Meloidogyne hapla and Glomus fasciculatum on Allium cepa (onion) grown in organic soil was evaluated under greenhouse conditions. In the absence of G. fasciculatum, M. hapla significantly retarded the growth of A. cepa cv. Krummery Special and MSU 8155 x 826, but had no detrimental influence on Downing Yellow Globe, Spartan Banner, or Spartan Sleeper. All five cultivars maintained populations of M. hapla, Final root population densities of M. hapla associated with Spartan Banner, Krummery Special, MSU 8155 x 826, and Spartan Sleeper were significantly greater than those recovered from Downing Yellow Globe. Final root population densities of M. hapla were directly proportional to the initial population densities. Root colonization of onion by G. fasciculatum significantly enhanced the growth and development of Downing Yellow Globe. The rate of increase of A. cepa growth and the final spore density were directly proportional to the initial spore density of G. fasciculatum. Final population densities of M. hapla in the presence of G. fasciculatum were generally greater than in the absence of the fungus. After 15 weeks, A. cepa plants grown in the presence of both M. hapla and G. fasciculatum were significantly larger than those grown in the presence of only M. hapla. PMID- 19294059 TI - Ethylene Production by Meloidogyne spp.-Infected Plants. AB - Gall size and rates of ethylene production by various hosts infected with Meloidogyne javanica and by excised tomato root cultures infected with M. javanica or M. hapla were measured. Infection with M. javanica increased the rate of ethylene production in dicotyledonous plants (cabbage, pea, carrot, cucumber, carnation, and tomato), but not in infected monocotyledonous plants (corn, wheat, and onion). Nematode infection induced large galls on roots of dicotyledonous, but not monocotyledonous, plants. Excised tomato roots in culture infected with M. javanica produced ethylene at high rates and formed large galls, whereas roots infected with M. hapla produced ethylene at low rates and induced smaller galls. PMID- 19294060 TI - Sterol Composition of the Corn Cyst Nematode, Heterodera zeae, and Corn Roots. AB - Sterols from free sterol and steryl ester fractions from Heterodera zeae and from total lipids of Zea mays roots were analyzed by gas-liquid chromatography (GLC) and by GLC-mass spectrometry. The major free sterols of H. zeae were 24 ethylcholesterol (54.4% of total free sterol), 24-ethylcholesta-5,22-dien-3beta ol (13.3%), 24-methylcholesterol (12.5%), and cholesterol (7.2%). The same four sterols comprised 34.6%, 7.2%, 30.3%, and 18.6%, respectively, of the esterified sterols of H. zeae. Corn root sterols included 46.6% 24-ethylcholesta-5,22-dien 3beta-ol, 16.7% methylcholesterol, 16.4% cycloartenol, 12.7% 24-ethylcholesterol, and 0.5% cholesterol. The sterol 24-composition of H. zeae differed greatly from that of the only other cyst nematode previously investigated, Globodera solanacearum. PMID- 19294061 TI - Population Development and Influence of Bursaphelenchus xylophilus on Gliocladium virens. AB - Gliocladium virens was isolated from slash pine trees symptomatic and asymptomatic for pine wilt disease with frequencies of 24% and 10%, respectively. Populations of Bursaphelenchus xylophilus, the nematode incitant of this disease, reproduced on this fungus and inhibited its growth. Growth inhibition of the fungus was characterized by an absence of sporulation and by the formation of chains of dark, thick-walled, chlamydospore-like cells. Population increase during a 12-day period following infestation of cultures of the fungus with 10,000 nematodes averaged 3-fold at 16 C, 9-fold at 20 C, and 24-fold at 24 C. In greenhouse studies, nematode recovery from slash pine seedlings coinoculated with both organisms was significantly greater than that obtained from seedlings inoculated with the nematode alone. PMID- 19294062 TI - Host Suitability and Reaction of Bean and Pea Cultivars to Meloidogyne chitwoodi and M. hapla. PMID- 19294063 TI - Pathogenicity of Pratylenchus penetrans to Navy Bean (Phaseolus vulgaris L.). AB - The pathogenicity of Pratylenchus penetrans (root-lesion nematode) to Phaseolus vulgaris (navy bean) was evaluated in greenhouse experiments. Shoot and root fresh weight of cv. Sanilac plants were increased 4 and 21%, respectively, by an initial population density (Pi) of 25 P. penetrans per 100 cm(3) soil. Leaf area and shoot fresh and dry weights were decreased by a Pi of 50 or more P. penetrans per 100 cm(3) soil. A significant positive linear relationship existed between initial soil population densities of P. penetrans and final soil and root population densities of this nematode. Three dry bean cultivars, Sanilac, Seafarer, and Tuscola, were susceptible to P. penetrans, and yields were reduced by 43-76% when plants were exposed to a Pi of 150 P. penetrans per 100 cm(3) soil. P. penetrans also reproduced on bean cultivars Saginaw, Gratiot, and Kentwood, but did not decrease bean yields, suggesting that these cultivars were tolerant to this nematode. PMID- 19294064 TI - Spatial pattern analysis of plant-parasitic nematodes. AB - Spatial patterns of Meloidogyne incognita, Tylenchorhynchus claytoni, Helicotylenchus dihystera, and Criconemella ornata were analyzed using Hill's two term local quadrat variance method (TTLQV), spectral analysis, and spatial correlation. Data were collected according to a systematic grid sampling plan from seven tobacco fields in North Carolina. Different estimates of nematode cluster size were obtained through TTLQV and spectral analysis. No relationship was observed between either estimate and nematode species, time of sampling (spring vs. fall), or mean density. Cluster size estimates obtained from spectral analysis depended on sampling block size. For each species examined, spatial correlations among nematode population densities were greater within plant rows than across rows, indicating that clusters were ellipsoidal with long axes oriented along plant rows. Analysis of mean square errors indicated that significant gains in sampling efficiency resulted from orienting the long axis of sampling blocks across plant rows. Spatial correlation was greater in the fall than in spring and was greater among 1 x 1-m quadrats than among 3 x 3-m quadrats. PMID- 19294065 TI - Density-dependent nematode seasonal multiplication rates and overwinter survivorship: a critical point model. AB - Nematode multiplication rates Pf/Pi and overwinter survivorship (Pi2/Pfl) for Meloidogyne incognita were both adequately described by negative exponential models, indicating density dependence in each case. Density dependence of the multiplication rates is mediated by resource limitation and host damage; in survivorship rates it may be mediated by limitation of stored reserves or prevalence of antagonists. Parameters of multiplication rate models were crop specific and varied with host status and environmental suitability. Maximum multiplication rates (a) of nearly 1,000 were measured for tomatoes. Equilibrium densities were sensitive to tolerance of the nematode by the crop. Overwinter survival rates varied among locations where cultural practices and length of infestation time differed. PMID- 19294066 TI - Population Trends and Vertical Distribution of Plant-Parasitic Nematodes Associated with Vitis labrusca L. in Michigan. AB - Nematode population trends and vertical distribution were monitored in a southwest Michigan vineyard (Vitis labrusca cv. Concord) from 1976 through 1983. Shallow (20 cm) and deep (90 cm) applications of 1,3-dichloropropene applied at 281 (shallow) plus 658 or 1,122 (deep) liters/ ha provided excellent control of Xiphinema americanum, Criconemella xenoplax, and Meloidogyne hapla. Populations of X. americanum remained below detectable levels for the entire 8-year experimental period where the fumigant was applied. X. americanum and C. xenoplax populations exhibited multiyear cycling in nonfumigated plots. M. hapla was first detected in 1978 and increased in prominence from 1980 through 1982. Criconemella spp. were commonly parasitized by an endoparasitic fungus. Parasitism was monitored and reported as an indication of nematode population quality. PMID- 19294067 TI - Description of Pratylenchoides sheri n. sp. (Nematoda: Pratylenchidae). AB - Pratylenchoides sheri n. sp., a bisexual species from the Pacific coastal area of California, is described and illustrated. Its most distinctive characters are the esophageal glands slightly overlapping the intestine dorsally, the large ventrally slanting esophago-intestinal valve, the lack of labial sectors in the first lip annule, the presence of males, and apparently functional spermatheca. Pratylenchoides sheri is morphologically similar to P. magnicauda, but differs in length and labial conformation, its more anterior vulva, and the presence of males. Relationships of head and esophagus gland lobe characters of P. sheri are compared to other Pratylenchoides species. PMID- 19294068 TI - Ultrastructure of the Head Region of Molting Second-Stage Juveniles of Heterodera glycines with Emphasis on Stylet Formation. AB - The morphology and alterations of infective juvenile (J2) body components with emphasis on the body wall, stomatal wall, stylet, and sensilla of Heterodera glycines were observed. During the molt of J2 to J3, the J2 hypodermis separates from the J2 cuticle and forms an extracellular space, continuous with an invagination of the anterior, center of the J3. The space between the J2 cuticle and the enlarged J3 hypodermal cells is filled with electron-dense material resembling a fluid observed in insects during molt. Regeneration of the J2 during molt was traced in a series of ultrathin sections. The site of stylet regeneration is in the hypodermal and myoepithelial tissues of the invaginated anterior, center of the J3. Four arcade-like cells are related to specific components of the stomatal wall, the stylet cone, and the stylet shaft of the J3. The first and second arcade-like cells are primarily related to stomatal wall development, whereas the third and fourth arcade-like cells are related to stylet cone and shaft development. Spherical, electron-translucent vacuoles that occur in myoepithelial cells just posterior to the arcade-like cells appear to be progenitors of the stylet knobs. Early stages of protractor muscle attachment to the vacuolar membrane were observed. PMID- 19294069 TI - Root Penetration by Meloidogyne incognita Juveniles Infected with Bacillus Penetrans. AB - Bacillus penetrans inhibited penetration by Meloidogyne incognita second-stage juveniles (J2) into tomato roots in the laboratory and greenhouse. Spores from this Florida population of B. penetrans attached to J2 of M. javanica, M. incognita, and M. arenaria. A greater proportion of J2 of M. javanica were infected than were J2 of either M. incognita or M. arenaria, and a greater number of spores attached to M. incognita than to M. arenaria. PMID- 19294070 TI - Nematode community structure in dogwood, maple, and peach nurseries in tennessee. AB - Nursery blocks (48 dogwood, 27 maple, 17 peach) in 20 middle Tennessee nurseries were sampled for nematodes in March,July, and October 1981. Dogwoods and maples were grouped in three age classes: 1-2, 3-5, and 10+ years. Nematodes were extracted from soil samples, counted, and assigned to trophic groups as follows: plant parasites, microbivores, fungivores, predators, and omnivores (= Dorylaimida). Total nematode numbers per 200 cm s soil ranged from 52 to 9,166 (mean = 1,785 +/- 1,420). Nematodes were more abundant in dogwood and maple than in peach blocks, and their numbers were significantly correlated with percentage of weed ground cover and number of weed species. Nematode numbers in dogwood sites were also correlated with dogwood age. Microbivores were the most abundant trophic group in all sites, followed by plant parasites, fungivores, omnivores, and predators. Nematode communities in nursery sites shared characteristics of both undisturbed and agricultural habitats. Degree and diversity of plant ground cover appeared to be the most important factors determining nematode community structure. PMID- 19294071 TI - Plant-parasitic nematode communities in dogwood, maple, and peach nurseries in tennessee. AB - Nursery blocks (48 dogwood, 27 red maple, and 17 peach) distributed among 20 Tennessee nurseries were sampled for nematodes in March, July, and October 1981. Plant-parasitic nematodes were extracted from soil, counted by genera, and identified to species after fixation. A total of 57 species in 24 genera were found, with 1-16 species occurring in a site. The species most commonly detected were Paratylenchus projectus and Xiphinema americanum, which were found in 88% and 78% of the sites, respectively. Relationships existed between distribution and densities of some species present in more than 10% of the sites and certain soil factors (pH, bulk density, texture, and organic matter content). Plant parasitic nematode community diversity was related to tree age, percentage of weed ground cover, and number of weed species. Site similarities in community ordinations were dependent on the individual nurseries sampled, tree age, and soil type, but clusters of sites of similar tree ages and soil types were not exclusive. PMID- 19294072 TI - Effect of Ditylenchus dipsaci on Alfalfa Mortality, Winterkill, and Yield. AB - Ditylenchus dipsaci-infected and noninfected alfalfa plants in a naturally infested field were studied from July 1980 to September 1982. Forty-one percent of the plants died during the study. Ninety-seven percent of the plants that died were infected with D. dipsaci. Sixty-nine percent of the observed mortality occurred during winter. Forage yield of infected plants was significantly lower than yield of noninfected plants at each harvest. Stored carbohydrates in infected plants were significantly lower than in noninfected plants. In a controlled environment test, significantly greater mortality occurred in frozen severely infected plants than in frozen noninfected plants, while no mortality occurred in severely infected or noninfected plants that were not frozen. Both forage yield and stored carbohydrates were significantly lower in severely infected than noninfected, non-frozen plants. Mortality in greenhouse-grown plants that were transplanted to field plots was significantly greater in D. dipsaci-infected plants than in noninfected plants after one winter. PMID- 19294073 TI - Effect of Inhibitors and Stimulators of Ethylene Production on Gall Development in Meloidogyne javanica-Infected Tomato Roots. AB - Excised tomato roots infected with Meloidogyne javanica produced ethylene at 3-6 times the rate of noninfected roots. This increase in ethylene production started 5 days after inoculation. Gall growth and ethylene production in infected roots were accelerated by 1-aminocyclopropane-1-carboxylic acid (ACC), indole acetic acid (IAA), and ethrel known as ethylene production stimulators. When inhibitors of ethylene production, like aminoethoxyvinylglycine (AVG) or aminoxyacetic acid (AOA), or inhibitors of ethylene action like silver thiosulfate (STS), were applied, gall growth and ethylene production were inhibited. Enhanced expansion of parenchymatous cells was observed in sections from nematode-induced galls and ethylene-treated roots. Lignification of xylem elements and fibers in the vascular cylinder was markedly inhibited in the gall, compared with noninfected root tissue. Because ethylene is known to induce cell expansion and to inhibit lignification, it is suggested that this plant hormone plays a major role in the development of M. javanica-induced galls. Ethylene affects gall size by enhancing parenchymatous tissue development and allows expansion of giant cells and the nematode body by reducing tissue lignification. PMID- 19294074 TI - Heterorhabditis sp. (Nematoda: Heterorhabditidae): A Nematode Parasite Isolated from the Banded Cucumber Beetle Diabrotica balteata. AB - A nematode identified as Heterorhabditis sp. was discovered in June 1982 in larval cadavers of the banded cucumber beetle, Diabrotica balteata, in soil on wooded land. Effective beetle control (over 95%) was obtained when larvae were exposed to potted soil containing infective stage nematode juveniles or infected larval cadavers. The nematode was propagated in vivo on larvae of D. balteata, Diaphania nitidalis (the pickleworm), and Galleria mellonella (the greater wax moth). This Heterorhabditis sp. has promising potential as a biocontrol agent for the banded cucumber beetle. PMID- 19294075 TI - Population Dynamics of Heterodera glycines Life Stages on Soybean. AB - Population fluctuations of Heterodera glycines differ in fields with high and low initial population densities. In a field with low initial numbers of nematodes, the numbers of cysts and eggs in soil remained low through 100 days from planting then increased during the remainder of the growing season. In a field with high initial nematode populations, numbers increased at 30 days, decreased to low numbers at 100 days, and then resurged to maximum populations at harvest. Numbers of juveniles were greatest at 100 days in the low initial population density field and at planting in the high initial population density field. The initial numbers of eggs in the soil gave the best correlation to soil and root nematode populations 15 and 30 days later. Juveniles in the soil at planting gave the largest correlation coefficients with nematode populations in the roots at 15 days in the field with the low initial population density. Eggs and juveniles in the soil at harvest were poorly related to numbers that overwintered. PMID- 19294076 TI - Occurrence of Biotypes in Radopholus citrophilus. AB - Two morphologically and karyotypically identical populations of the citrus burrowing nematode, Radopholus citrophilus, differed in their ability to damage and reproduce in roots of citrus rootstocks previously identified as either resistant or tolerant. These populations are considered to be biotypes, and their occurrence may explain the appearance of spreading decline symptoms in plantings of rootstocks previously considered resistant. PMID- 19294077 TI - Studies on the Host Range, Biology, and Pathogenicity of Punctodera punctata Infecting Turfgrasses. AB - Punctodera punctata completed its life cycle on Poa annua (annual bluegrass), P. pratensis (Merion Kentucky bluegrass), Lolium perenne (perennial ryegrass), and Festuca rubra rubra (spreading fescue). Minimum time for completion of a life cycle from second-stage juvenile to mature brown cyst was 40 days at 22-28 C. Inoculation by single juveniles indicated that reproduction was most likely by amphimixis. Infestation levels of 50 or 500 juveniles/250 cm(3) soil did not affect top dry weight, root dry weight, or total dry weight of Poa annua. PMID- 19294078 TI - Responses of Anguina agrostis to Detergent and Anesthetic Treatment. AB - The infective dauer juvenile (DJ2) of Anguina agrostis, a stage capable of surviving desiccation, is up to sixfold more resistant to the detergent sodium dodecyl sulfate than are freshly hatched juveniles or adult males, and twofold more resistant to the anesthetic phenoxypropanol. Thus, the DJ2, like dauer stages of other species, may also be more resistant to various types of environmental stress in its natural habitat. In A. agrostis, however, resistance appears to be acquired gradually during development of the second juvenile stage, rather than during a molt. PMID- 19294079 TI - Effects of Pratylenchus zeae and Quinisulcius acutus Alone and in Combination on Sorghum. AB - Host-parasite relationships of Pratylenchus zeae and Quinisulcius acutus, alone or in combination, were studied on sorghum in the greenhouse and laboratory. Q. acutus at 1,000 or 5,000 nematodes per 15-cm-d pot and P. zeae at 500 nematodes per pot significantly suppressed plant height and fresh and oven dry shoot and root weights. A mixture of 1,000 Q. acutus and 500 P. zeae per pot resulted in greatest suppression of growth. Roots of plants inoculated with Q. acutus alone were reduced in number and size and showed lesions and discoloration. Reproduction of this nematode 42 days after inoculation was much greater in treatments of 100 or 1,000 than 5,000 nematodes. The population density of the two species at 6 weeks after inoculation was significantly less when combined than for each species alone. When the two species were combined, reproduction of P. zeae was greater than that of Q. acutus, but the final populations per gram of root weight were the same. Q. acutus fed ectoparasitically on epidermal cells of sorghum roots in the zone of elongation and differentiation when observed under in vitro conditions. PMID- 19294080 TI - Ultrastructure of the Phasmids of Scutellonema brachyurum. AB - Electron microscopical studies reveal that the phasmids of Scutellonema brachyurum include socket and sheath cells which appear to be secretory. The phasmid includes an external cup-shaped ampulla filled with electron-dense material which may be lipopolysaccharide. The ampulla is continuous, with a receptor cavity surrounded by the socket and sheath cells. The dendrite receptor originates from a neuron and directs posteriorly through a receptor socket into the receptor cavity ending near the electron-dense plug. Serial sections through the dendrite receptor indicate that it does not conform to the typical 9+2 arrangement of microtubules characteristic of cilia in higher animals. PMID- 19294081 TI - Cactodera eremica n. sp., Afenestrata africana (Luc et al., 1973) n. gen., n. comb., and an Emended Diagnosis of Sarisodera Wouts and Sher, 1971 (Heteroderidae). AB - Systematic contributions to Heteroderidae include description of Cactodera eremica n. sp., an emended diagnosis of Sarisodera Wouts and Sher, 1971, and proposal of a new genus and new combination, Afenestrata africana (synonym Sarisodera africana Luc et al., 1973). Cactodera eremica, from the roots of shadscale in Utah, most closely resembles Cactodera thornei (Golden and Raski, 1977) but differs by the presence of a finely striated cuticle, a fine surface pattern on eggs, a shorter female stylet, distance of the DGO from the stylet, vulval slit, and smaller diameter of circumfenestra, as well as a shorter tail in second-stage juveniles. The response of the host to C. eremica is similar to other Heterodera sensu lato including a large syncytium with wall ingrowths. The diagnosis of Sarisodera is emended to exclude cysts, which do not form in the type species, S. hydrophila. Afenestrata africana differs from S. hydrophila by the formation of cysts, the dorsal position of the anus in females, the shorter stylet, and a pore-like phasmid opening in second-stage juveniles. In addition, the lip pattern of males and juveniles is characterized by a greater degree of fusion of lip parts, the host response is a syncytium (versus a single uninucleate giant cell in S. hydrophila), and the cuticle is thinner and lacks a D layer. Unlike Heterodera, the cyst of Afenestrata lacks fenestrae. PMID- 19294082 TI - Emendation of the Genus Paurodontella Husain &Khan, 1968 and Description of P. auriculata n. sp. from Western Canada (Nematoda: Paurodontidae). AB - Males and females of Paurodontella auriculata n. sp. are described and illustrated and the genus emended. The species is characterized by a short, robust body of 355-525 mum, a stylet length of 7-8 mum, lateral field with 5-6 incisures, and a conoid, attenuated tail 1.5 times the vulva-anus distance. Primary characters used in emending the genus are asymmetrical stylet knobs, absence of lateral lips, a deep slit-like amphid dividing the submedian lips to base of head, where open lateral sides are covered by a large auriform cuticular flap, and variable length of the basal esophageal bulb stem and male bursa. PMID- 19294083 TI - Description of Meloidogyne pini n. sp., a Root-Knot Nematode Parasitic on Sand Pine (Pinus clausa), with Additional Notes on the Morphology of M. megatyla. AB - Meloidogyne pini n. sp. is described from sand pine, Pinus clausa, in Georgia. The perineal pattern of the female has a large cuticular ridge surrounding a deeply recessed perivulval area. The lateral fields are marked by transverse striae. The female stylet is 14.6 mum long, and the knobs are small, rounded, and set off from the straight and narrow shaft. The excretory pore is near the level of the base of the stylet. The labial disc of the male is large, rounded, and fused with the crescent-shaped medial lips. The head region is smooth, the styler is 20.8 mum long, and the cone is more than twice as long as the shaft. The knobs are rounded and set off from the shaft. In the second-stage juvenile, the labial disc, medial lips, and lateral lips form one smooth, continuous, ovoid head cap. Mean juvenile length is 434 mum, stylet length is 12.8 mum, and tail length is 44.4 mum. M. pini n. sp. also parasitizes loblolly and slash pine. Additional morphological details of M. megatyla are presented. PMID- 19294085 TI - Life History of Pratylenchus vulnus on Carrot Discs. PMID- 19294084 TI - Catalogue of types deposited in the canadian national collection of nematodes. AB - Primary and secondary types housed in the Canadian National Collection of Nematodes are given for 396 species of plant-parasitic, marine, freshwater, terrestrial, and entomophagous nematodes. Species are listed in alphabetical order and include author(s), date, publication source, type category, numbers and sex, and collection accession number. Current nomenclatorial changes in status of a binomial are not given, but authority data are added for type designations made to the original type series. PMID- 19294086 TI - Criconemella anastomoides n. sp. (Nematoda: Criconematina) from Pakistan. AB - Criconemella anastomoides, described and illustrated herein, is characterized by the presence of regular anastomosis on the body, two zig zag lateral lines, stylet knobs sloping posteriorly, short stylet, and short body length. PMID- 19294087 TI - Observations of Molting and Population Development by Orrina phyllobia. PMID- 19294088 TI - Anatomical Changes Induced by Punctodera chalcoensis in Corn Roots. PMID- 19294089 TI - Overestimation of Yield Loss of Tobacco Caused by the Aggregated Spatial Pattern of Meloidogyne incognita. AB - Overestimation of yield loss caused by Meloidogyne incognita on tobacco was calculated as a function of the statistical frequency distribution of sample counts. Sampling frequency distributions were described by a negative binomial model, with parameter k, and the resulting probability generating function was used to calculate discrete damage probabilities. Negative binomial damage predictions were compared to mean-density estimates of damage. Predictions based on mean density alone overestimate yield loss by values ranging from 300% at a k of 0.1 to less than 10% at a k of 1.0. Damage overestimation was described as an exponential function of k and mean density. Preplant sampling data for M. incognita were used to derive a linear model for the estimation of k from mean density, allowing the calculation of yield-loss overestimation based on one parameter, the field mean density. Overestimation of damage ranged from 288% at a density of 50 juveniles/500 cm(3) soil, to 5% at a density of 1,000 juvelfiles/500 cm(3) soil. PMID- 19294090 TI - Dynamics of Winter Survival of Eggs and Juveniles of Meloidogyne incognita and M. arenaria. AB - Winter survival dynamics of Meloidogyne incognita and M. arenaria were studied at nine sites in Texas for 2 years. Population survival from October until April was variable among sites, ranging from 0.1% to 33%. A negative correlation (r = 0.86, P = 0.01) was observed between initial population, densities in October and survival percentage until the following April. Total population (eggs + J2) and population of eggs declined continuously during the survival period. Populations of juveniles (J2) increased initially, then declined. The total populations were 82% eggs in October; hatch of these eggs was believed responsible for the observed increase in the population density of J2. Viable eggs were recovered from the soil until March. Eggs are as important as J2 in winter survival of M. incognita and M. arenaria in Texas. Survival data were analyzed by a simple mathematical model. PMID- 19294091 TI - A Theoretical Model of the Winter Survival Dynamics of Meloidogyne spp. Eggs and Juveniles. AB - A theoretical model of the winter survival dynamics of Meloidagyne spp. was developed by considering the roles of egg hatching and juvenile mortality and the initial populations of eggs and juveniles at the onset of winter. Two distinct patterns of juvenile dynamics appear which depend upon the numerical values of the model parameters. The model predicts whether eggs or juveniles are the major component of overwintering nematode populations at any time prior to planting. The model could be elaborated to include egg viability and differential mortality of eggs but at some cost in ease of mathematical analysis. A general procedure for fitting the proposed model to experimental or observational data is outlined. PMID- 19294092 TI - Permeability of the Body Wall of Romanomermis culicivorax to Lanthanum. AB - Ultrastructural study of the body wall of preparasitic, parasitic, and postparasitic stages of Romanomermis culicivorax showed that the cuticle of all three stages was permeable to lanthanum. The cuticle of the parasitic stage was the thinnest and showed the greatest permeability. Lanthanum accumulated on the apical surfaces of the hypodermal cells but was not found intracellularly. The negative staining characteristics of lanthanum enhanced the detection of numerous smooth septate junctions in the hypodermis of the parasitic stage. PMID- 19294093 TI - Leucine Aminopeptidase in Eggs of the Soybean Cyst Nematode Heterodera glycines. AB - Supernatant from a sonicated macerate of eggs of Heterodera glycines hydrolyzed L leucine beta-naphthylamide and L-leucine 7-amido-4-methylcoumarin. Rate of substrate hydrolysis was influenced by pH and increased with the duration of incubation. A Michaelis-Menten constant of 0.15 mM was obtained. Rate of substrate hydrolysis was decreased by freezing egg supernatant for 26 days or heating above 60 C for 5 minutes. When egg supernatant was incubated with six different substrates, L-leucine beta-naphthylamide was hydrolyzed most readily and L-valine beta-naphthylamide the least readily. The rate of substrate hydrolysis by egg supernatant was not increased by pretreatment of eggs with 3 mM zinc chloride for up to 14 days. PMID- 19294094 TI - Plant-Induced Hatching of Eggs of the Soybean Cyst Nematode Heterodera glycines. AB - Root diffusate from soybean plants caused greater hatching of Heterodera glycines eggs during vegetative growth of the host, but the activity declined with plant senescence. Chelation of the root diffusate with ethylenediamine tetraacetic acid (EDTA) significantly increased hatching activity for H. glycines eggs. Diffusate from leafless plants caused little hatching, whereas treatment of intact plants with the growth regulators gibberellin and kinetin had no effect on the hatching activity of root diffusate. Treating H. glycines eggs with zinc chloride and root diffusate reduced egg hatching from zinc chloride alone. Levels of zinc in the root diffusate were insufficient to induce egg hatch, based on analysis by atomic absorption spectrophotometry. The enzymatic activity of leucine aminopeptidase in H. glycines eggs was not altered by treatment with chelated or nonchelated root diffusate. PMID- 19294095 TI - Morphological Comparisons of Host Races 1 and 2 of Meloidogyne arenaria from Florida. AB - Morphometric comparisons of two populations each of Races 1 and 2 of Meloidogyne arenaria from tobacco, peanut, and soybean in different localities in Florida showed no significant differences in any characters measured by light microscopy. Thirteen morphometric characters of females, 22 of males, and 21 of infective second-stage juveniles were measured for each population. The stylet length in each population and in each life stage was the least variable character. Proposed character additions to supplement the description of M. arenaria and more precisely define the species include the following: female - metacorpus length and width, metacorpus valve length and width, vulva-anus distance; male - length of stylet shaft and knobs, stylet base to head end; infective juvenile - length of stylet shaft and knobs, head end to metacorpus valve, excretory pore to head end, metacorpus valve length and width, and d-ratio. PMID- 19294096 TI - Fine structure of body wall cuticle of females of eight genera of heteroderidae. AB - Body wall cuticle of adult females of eight genera within the Heteroderidae was examined by transmission electron microscopy for comparison with previously studied species within the family. Cuticle structure was used to test some current hypotheses of phylogeny of Heteroderidae and to evaluate intrageneric variability in cuticle layering. Verutus, Rhizonema, and Meloidodera possess striated cuticle surfaces and have the simplest layering, suggesting that striations have not necessarily arisen repeatedly in Heteroderidae through convergent or parallel evolution. Atalodera and Thecavermiculatus possess similar cuticles with derived characteristics, strengthening the hypothesis that the two genera are sister groups. Similarly, the cuticle of Cactodera resembles the specialized cuticle of Globodera and Punctodera in having a basal layer (D) and a surface layer infused with electron-dense substance. Heterodera betulae has a unique cuticle in which the thickest layer (C) is infiltrated with an electron dense matrix. Little intrageneric difference was found between cuticles of two species of Meloidodera or between two species of Atalodera. However, Atalodera ucri has a basal layer (E) not found in other Heteroderidae. The most striking intrageneric variation in cuticle structure was observed between the thin three layered cuticle of Sarisodera africana and the much thicker four-layered cuticle of Sarisodera hydrophila; results do not support monophyly of Sarisodera. PMID- 19294097 TI - Comparative Morphology of Meloidodera spp. and Verutus sp. (Heteroderidae) with Scanning Electron Microscopy. AB - Scanning electron microscopy (SEM) of second-stage juveniles (J2), males, and females of Meloidodera floridensis, M. charis, M. belli, and Verutus volvingentis reveals detailed characteristics of the head region, lateral field, phasmid, body striae, vulva, and perineal region. In M. charis and M. belli the en face pattern conforms to a basic pattern in which the labial disc is surrounded by six lips (sectors) of the first head annulation. In J2 the head has additional annulations, whereas in males annulation is replaced by longitudinal blocks. Conversely, J2 and males of M. floridensis and V. volvingentis each have a unique derived face pattern with fusion of various lip components and with head annulation. All six lips of females of M. charis and M. belli are fused, whereas females of M. floridensis and V. volvingentis have distinct lateral lips. Lateral fields vary among species, with only slight differences at the anterior and posterior ends of the lateral lines and in the spatial relation of the lines to phasmid openings. Phasmid openings are present in adults of Meloidodera spp., but were not observed in adults of V. volvingentis; in this respect, the female perineal pattern of Verutus is different from Meloidodera spp, The very large vulva (+/- 48 mum long) of V. volvingentis is in sharp contrast to the minute vulva (+/- 6 mum long) in a population of M. charis from San Bernardino. Morphological characters revealed by SEM will be most informative when investigated throughout Heteroderidae and incorporated with additional characters for a phylogenetic analysis of the family. PMID- 19294098 TI - Xiphidorus amazonensis n. sp. (Nematoda: Longidoridae) from the Brazilian Amazon Basin. AB - Xiphidorus amazonensis n. sp. was found in the rhizospheres of Jatropha curcas, Musa sp., Anona muricata, Cassia tora, Panicum laxum, Paspalum fasciculatum, Aeschynomene sensitiva, Saccharum officinarum, Manihot esculenta, Abelmoschus esculentus, Tamarindus indica, Mangifera indica, Vigna unguiculata, Zea mays, Commelina sp., Cyperus rotundus, Fimbristylis miliacea, Citrus sinensis, and Eichhornia crassipes on the Amazon River island of Xiborena, approximately 40 km southeast of Manaus, capital of the State of Amazonas. The type habitat is flooded annually for about 6 months by the Amazon River. Xiphidorus amazonensis n. sp. differs from the closely related species Xiphidorus yepesara Monteiro, 1976 by the larger size, by a, b, and c values, and by the rounded tail terminus. It also resembles Xiphidorus tucumanensis Chaves and Coomans, 1984, but can be distinguished by its larger size, larger a, b, and c values, more conical female tail, bilobed amphidial pouch, and the presence of a spermatheca full of sperm. PMID- 19294099 TI - Nematode control related to fusarium wilt in soybean and root rot and zinc deficiency in corn. AB - Nematode and disease problems of irrigated, double-cropped soybean and corn, and zinc deficiency of corn were investigated. Ethylene dibromide, phenamiphos, and aldicarb were equally effective for controlling nematodes and increasing yields of corn planted minimum-till and soybean planted in a moldboard plow prepared seedbed. The residual effects on yields of nematicides applied to the preceeding crop occurred during 3 years for soybean and 1 year for corn. Fusarium wilt symptoms of soybean that developed during 2 years of the study were less severe in all nematicide-treated plots than in control plots. Typical zinc deficiency symptoms on 30-day-old corn plants were observed during 1 year of the study in certain plots. Symptoms were not evident on plants grown on plots treated with ethylene dibromide, and only occasional plants had symptoms on plots treated with phenamiphos and aldicarb. The amount of yield response directly related to nematode control could not be determined because of the apparent interaction of nematodes on the expression of Fusarium wilt of soybean. Our study strongly indicates that the expression of Fusarium wilt of soybean and zinc deficiency in corn are influenced by nematodes and that nematicides will reduce their severity. PMID- 19294100 TI - Development of Meloidogyne chitwoodi on Wheat. AB - Postinfection development of Meloidogyne chitwoodi from second-stage juveniles (J2) to mature females and egg deposition on 'Nugaines' winter wheat required 105, 51, 36, and 21 days at 10, 15, 20, and 25 C. At 25 C, the J2 induced cavities and hyperplasia in the cortex and apical meristem of root tips with hypertrophy of cortical and apical meristem cell nuclei, 2 and 5 days after inoculation. Giant cells induced by late J2 were observed in the stele 10 days after inoculation. Clusters of egg-laying females were common on wheat root galls 25 days after inoculation. Juveniles penetrated wheat roots at 4 C and above, but not at 2 C, when inoculum was obtained from cultures grown at 20 C, but no penetration occurred at 4 C when inoculum was stored for 12 hours at 4 C before inoculation. In northern Utah, J2 penetrated Nugaines wheat roots in the field in mid-May, about 5 months after seedling emergence. M. chitwoodi eggs were first observed on wheat roots in mid-July when plants were in blossom. Only 40% of overwintered M. chitwoodi eggs hatched at 25 C. PMID- 19294101 TI - Control of Root-Knot Nematodes on Tomato by the Endoparasitic Fungus Meria coniospora. AB - The endoparasitic nematophagous fungus Meria coniospora reduced root-knot nematode galling on tomatoes in greenhouse pot trials. The fungus was introduced to pots by addition of conidia at several inoculum levels directly to the soil or addition of nematodes infected with M. coniospora to the soil; both methods reduced root galling by root-knot nematodes. These studies represent a part of a recently initiated effort to evaluate the potential of endoparasitic nematophagous fungi for biocontrol of nematodes. PMID- 19294102 TI - Pratylenchus australis n. sp. and Eutylenchus fueguensis n. sp. (Nematoda: Tylenchina) from southern Chile. AB - Two new species of nematodes from southern Chile are described and illustrated. Pratylenchus australis n. sp. is distinguished by its heavy cephalic sclerotization, smooth tail terminus, lack of spermatheca, and absence of males. Eutylenchus fueguensis n. sp. differs from other Eutylenchus spp. by the long female stylet (31 [28-32] mum), strongly sclerotized excretory duct opening posterior to nerve ring, and broadly rounded caudal alae of males. PMID- 19294103 TI - Extraction of Cysts and Eggs of Heterodera schachtii from Soil with an Assessment of Extraction Efficiency. AB - Techniques are described for the extraction and enumeration of cysts and contained eggs from soil samples. The average recovery of cysts from seeded soil samples of differing soil texture was 82.7 +/- 2.1%. Recovery from sandy clay soil samples seeded at 1 cyst/100 g soil was 63.4 +/- 5.5%; at 4.2 cysts/100 g soil recovery was 89.6 +/- 1.8%. Recovery of cysts from naturally infested clay soil was 88.3 +/- 2.05%. Egg extraction efficiency for seeded samples was calculated as 78%, and for naturally infested soil was estimated as 83%. PMID- 19294104 TI - Parasitic and Saprophytic Abilities of the Nematode-Attacking Fungus Hirsutella rhossiliensis. AB - The ability of Hirsutella rhossiliensis to colonize various substrates in sterile and nonsterile soil was measured. Hirsutella rhossiliensis was recovered from 67% and 77% of living, inoculated Criconemella xenoplax incubated in sterile and nonsterile soil, respectively. In contrast, the fungus was recovered from 100% and 18% of heat-killed, inoculated nematodes incubated on sterile and nonsterile soil, respectively. Hirsutella rhossiliensis was readily recovered from inoculated, autoclaved wheat seeds incubated in sterile soil but not from seeds incubated in nonsterile soil. Autoclaved peach roots were a poor substrate for the fungus. Germination of H. rhossiliensis spores incubated on agar disks above soil was about 90% regardless of soil treatment. However, germ tube length was greatly suppressed by nonsterile soil. Our results suggest that H. rhossiliensis is a better parasite than saprophyte and that the fungus may be specialized for attacking nematodes. PMID- 19294105 TI - Sphaeronema sasseri n. sp. (Tylenchulidae), a Nematode Parasitic on Fraser Fir and Red Spruce. AB - Sphaeronema sasseri n. sp. is described from Fraser fir and red spruce on Mount Mitchell in North Carolina. Females are distinguished from other species in the genus by body shape, occurrence of body annulations, stylet morphology, head shape, and by several morphometric characters. The nematodes occur in colonies surrounding the bases of lateral and feeder roots, and the infected tissues show a general breakdown of the cortex and bark. The roots appear to be severely damaged by high populations of nematodes. This parasite may be important in the etiology of the slow decline of spruce and fir that has occurred in recent years in the southern Appalachian Mountains. PMID- 19294106 TI - Seasonal Fluctuations of Globodera tabacum solanacearum as Estimated by Two Soil Extraction Techniques. AB - Two soil extraction methods were compared to determine their efficiency in recovering cysts and juveniles of a tobacco cyst nematode, Globodera tabacum solanacearum. The methods were equally efficient when extracting nematodes from soil samples seeded in the laboratory; however, there was a significant extraction method x month interaction when the methods were used to estimate field soil populations over 2 years. The centrifugal sugar flotation method recovered greater numbers of cysts when densities were near 400 cysts/100 cm(3) soil and greater numbers of juveniles in all samples. The sugar flotation method recovered greater numbers of cysts during months when densities were less than 400 cysts/100 cm(3) soil. Numbers of cysts and juveniles were lowest in June and July following land tillage in May. A soil freeze in January 1982 may have been responsible for unusually high numbers of recovered cysts in February and March 1982, a pattern that did not occur in 1983. PMID- 19294107 TI - Hexamermis cathetospiculae n. sp. (Mermithidae: Nematoda), a Parasite of the Rice Stemborer, Tryporyza incertulas (Wlk.) (Pyralidae: Lepidoptera) in Malaysia. AB - Hexamermis cathetospiculae n. sp. (Mermithidae: Nematoda), a parasite of the rice stemborer, Tryporyza incertulas (Wlk.), is described. This mermithid was observed to control the stemborer in Malaysia. It is distinguished from other members of the genus by the shape of the vagina, the nearly straight spicules, and the form of the amphids. PMID- 19294108 TI - Susceptibility of the Carrot Weevil (Coleoptera: Curculionidae) to Steinernema feltiae, S. bibionis, and Heterorhabditis heliothidis. AB - Larvae, pupae, and adults of the carrot weevil (Listronotus oregonensis) were infected and killed by the three entomophagous nematodes (Steinernema feltiae, S. bibionis, and Heterorhabditis heliothidis) under controlled conditions. Third stage larvae were more susceptible than pupae or adults. S. feltiae and S. bibionis were the most aggressive nematode species, causing larval mortality after 24-48 hours in both continuous and 2-hour contact with nematode suspension. The nematodes multiplied sufficiently in all insects at all stages of development; however, production of infective-stage larvae per host cadaver was variable. PMID- 19294109 TI - Interactions between six warm-season legumes and three species of root-knot nematodes. PMID- 19294111 TI - Transfer of type specimens. PMID- 19294110 TI - Single Modified Food Cell Induced by Helicotylenchus pseudorobustus in Corn Roots. PMID- 19294112 TI - Host-Parasite Relationships of Meloidogyne arenaria and M. incognita on Susceptible Soybean. AB - Pathogenicity and reproduction of single and combined populations of Meloidogyne arenaria and M. incognita on a susceptible soybean (Glycine max cv. Davis) were investigated. Significant galling and egg mass production were observed on roots of greenhouse-grown soybean inoculated with M. arenaria and M. incognita, in combination and individually. M. arenaria produced more galls and egg masses than M. incognita, whereas in combined inoculation with both nematode species, gall and egg production was intermediate. In growth chamber tests, inoculations with M. arenaria and M. incognita, singly or in combination, produced more galls and egg masses at 30 C than at 25 C. At 25 C, M. arenaria alone produced significantly more galls and egg masses than the combined M. arenaria plus M. incognita, while M. incognita produced the fewest. At 30 C, numbers of egg masses produced by M. arenaria did not differ significantly from combined M. arenaria and M. incognita. In temperature tank tests, M. incognita produced more galls and egg masses at 28 C than at 24 C soil temperature. In contrast, numbers of galls, egg masses, and eggs of M. arenaria were slightly higher at 24 C than at 28 C. Combined inoculum of both nematode species produced greater numbers of galls at 24 C than at 28 C. PMID- 19294113 TI - Interrelationship of Heterodera schachtii and Meloidogyne hapla on Tomato. AB - Invasion of tomato (Lycopersicon esculentum L.) roots by combined and sequential inoculations of Meloidogyne hapla and a tomato population of Heterodera schachtii was affected more by soil temperature than by nematode competition. Maximum invasion of tomato roots, by M. hapla and H. schachtii occurred at 30 and 26 C, respectively. Female development and nematode reproduction (eggs per plant) of M. hapla was adversely affected by H. schachtii in combined inoculations of the two nematode species. Inhibition of M. hapla development and reproduction on tomato roots from combined nematode inoculations was more pronounced as soil temperature was increased over a range of 18-30 C and with prior inoculation of tomato with H. schachtii. M. hapla minimally affected H. schachtii female development, but there was significant reduction in the buildup of H. schachtii when M. hapla inoculation preceded that of H. schachtii by 20 days. PMID- 19294114 TI - Influence of Glomus fasciculatum on Meloidogyne hapla Infecting Allium cepa. AB - The impact of Glomus fasciculatum on Meloidogyne hapla associated with Allium cepa was evaluated in two experiments. Nematode density was not different in mycorrhizal and nonmycorrhizal plants 10 weeks after the joint inoculation of M. hapla and G. fasciculatum. Differences in the age structure of M. hapla populations reared on mycorrhizal and nonmycorrhizat plants were noted. G. fasciculatum enhanced leaf and bulb growth of A. cepa in the absence of M. hapla, but did not affect plant weight when nematodes were present. Survival and reproduction of M. hapla were not affected by G. fasciculatum or phosphorus (P). The estimated time required for inoculated second-stage juveniles (J2) to mature to the adult stage was 1,000 degree hours (base = 9 C) greater in mycorrhizal than in nonmycorrhizal plants supplemented with P. Although the infectivity of J2 was not measured directly, colonization of A. cepa by G. fasciculatum appeared to alter the ability of M. hapla to penetrate roots. PMID- 19294115 TI - Host-Parasite Relationship of Meloidogyne chitwoodi on Potato. AB - The soil fumigant 1,3-dichloropropene gave good to excellent control of the Columbia root-knot nematode, Meloidogyne chitwoodi, on potato, Solanum tuberosum L. Nonfumigant nematicides (aldicarb, fensulfothion, carbofuran, ethoprop, and phenamiphos) were less effective in controlling M. chitwoodi, since the nematode affects tuber quality more than quantity. Soil temperature during the growing season affected parasitism of M. chitwoodi on potato more than did the initial nematode population. There were positive linear correlations between degree-days and infected and galled tubers (r = 0.92), degree-days and nematode generations (r = 1.00), and infected and galled tubers and nematode generations (r = 0.91). Differences in degree-days and resultant nematode reproduction caused great variability in infection and galling of potato tubers during four growing seasons: 89% for 1979, 0% for 1980, 13% for 1981, and 18% for 1982, giving positive linear correlation (r = 0.99) between final nematode soil population (Pf) and percentage of infected and galled tubers. Corresponding increases in the soil populations of second-stage juveniles (J2) during the growing season were 9,700% in 1979, 170% in 1980,552% in 1981, and 326% in 1982. There was a negative linear correlation (r = -0.87) between initial soil J2 populations (Pi) and the degree of parasitism (infection and galling) of potato tubers, Pi being of secondary importance to degree-days. PMID- 19294116 TI - Selection and Inbreeding of Heterodera glycines on Glycine max. AB - Few soybean cyst nematodes (SCN), Heterodera glycines, of a diverse gene pool developed into females on soybeans PI 89772 or PI 209332. Nematodes surviving the selection pressure were then inbred for nine generations by single cyst transfers on the same selecting soybean line. These nematodes appeared to tolerate concurrent selection and inbreeding. Effects of selection-inbreeding, selection only, and secondary selection were evaluated by relative ability to produce cysts on 11 soybean lines. The genetic differences of PI 89772 (also Peking and Pickett 71) and PI 209332 were reaffirmed. The random effects of inbreeding indicated that Ilsoy and Williams may have genes for resistance different from those in PI 89772 or PI 209332. Egg inoculum obtained from soil resulted in very few cysts in some tests. Fresh egg inoculum (from cysts on 27-30-day-old plants) generally resulted in more cysts and more consistent results. Concurrent with the change in inoculum, there was a large increase in relative numbers of cysts on several soybean lines but no change on other lines; the true cause of this large interaction is unknown. Secondary selection of two inbreds was effective and suppressed cyst numbers on the line on which one inbred was selected initially. These results are consistent with the allelism linkage of some SCN genes reported previously. PMID- 19294117 TI - Corn response to subsoiling and nematicide application. AB - A 2-year field study evaluated the influence of subsoiling and nematicide application, alone and in combination, on the growth and yield of field corn in a sandy soil in north-central Florida. The field had a 25-30-cm-deep tillage pan (plowpan) and was infested with Belonolaimus longicaudatus, Hoplolaimus galeatus, Trichodorus christei, and Pratylenchus spp. Subsoiling increased corn yield both years, and the residual effect of subsoiling in the first year increased yields in the second year. Preplant application of DD injected in-row increased yields and reduced nematode populations. At-planting applications of DD injected in-row and carbofuran in-furrow or in a band were less effective than subsoiling in increasing yields and reducing nematode numbers. Interactions between subsoiling and nematicide treatments occurred in the second year. PMID- 19294118 TI - Influence of the Sting Nematode, Belonolaimus longicaudalus, on Young Citrus Trees. AB - The sting nematode, Belonolaimus longicaudatus, was associated with poor growth of citrus in a central Florida nursery. Foliage of trees was sparse and chlorotic. Affected rootstocks included Changsha and Cleopatra mandarin orange; Flying Dragon, Rubidoux, and Jacobsen trifoliate orange; Macrophylla and Milam lemon; Palestine sweet lime; sour orange; and the hybrids - Carrizo, Morton, and Rusk citrange and Swingle citrumelo. Root symptoms included apical swelling, development of swollen terminals containing 3-5 apical meristems and hyperplastic tissue, coarse roots, and a reduction in the number of fibrous roots. Population densities as high as 392 sting nematodes per liter soil were detected, with 80% of the population occurring in the top 30 cm of soil; however, nematodes were detected to 107 cm deep. Although an ectoparasite, the nematode was closely associated with citrus root systems and was transported with bare root nursery stock. Disinfestation was accomplished by hot water treatment (49 C for 5 minutes). PMID- 19294119 TI - Influence of Meloidogyne hapla on Alfalfa Yield and Host Population Dynamics. AB - Self-thinning in alfalfa, a dynamic process involving the progressive elimination of the weakest plants, was enhanced by Meloidogyne hapla. Alfalfa stand densities decreased exponentially with time and were reduced 62% (P = 0.05) in the presence of M. hapla. As stand densities decreased over time, mean plant weights increased at a rate 2.59 times faster in the absence of M. hapla. In a stepwise multiple regression analysis, 65% of the total variation in yield could be explained by changes in stand density and 85% by average weight of individual stems. Alfalfa yields were suppressed (P = 0.05) by M. hapla, with suppression generally increasing with time and as the nematode population density increased. Yield suppression was attributable primarily to the decline in plant numbers and to suppression in individual plant weights. PMID- 19294120 TI - Two-Dimensional Protein Patterns in Heterodera glycines. AB - Two-dimensional polyacrylamide gel electrophoretic protein patterns of H. glycines from southern Indiana (Posey County) and northern Indiana (Pulaski County) were largely similar, but many differences existed. The pattern of the Posey isolate was similar to patterns from isolates collected in other areas of the United States. Unique dense protein spots in the pattern of an isolate from Hokkaido, Japan, distinguished it from patterns of six U.S. isolates. PMID- 19294121 TI - Influence of Planting Date on Population Dynamics and Damage Potential of Pratylenchus brachyurus on Soybean. AB - Planting date was used as a variable to determine the effects of time and different environmental conditions on the population dynamics and damage potential of Pratylenchus brachyurus on soybean at two locations in North Carolina. An initial population slightly less than the damage threshold (275 nematodes/500 cm(3) soil) was used to minimize the influence of host damage on this nematode's population dynamics and to gain greater precision in characterizing factors which influence the damage potential of P. brachyurus to soybean. Equivalent nematode numbers generally resulted in greater yield suppression of soybean in early plantings. Early planting of soybean also resulted in greater (P = 0.01) population densities of P. brachyurus at midseason which often persisted until soybean harvest. Length of time for reproduction and intraspecific competition occurring when soybeans were stunted by the nematode were the most important factors influencing the population dynamics of P. brachyurus. PMID- 19294122 TI - Ultrastructure of Male Sexual Apparatus of Scutellonema brachyurum. AB - Electron micrographs of serial sections show that the male sexual apparatus of Scutellonema brachyurum includes two morphologically identical spicules. Each is composed of a swollen tubular head, crescentic shaft, and leaf-like blade with membranous velum expanded from the central trunk. The spicules are concave and grooved on the ventral side and convex on the dorsal side near the trunk. The trunk is continuous with the shaft and head. Nerve tissue occupies the core of the spicule and includes a dendritic process which gains access to the exterior via a small pore on the lateral side of the spicule tip. Three protractor and two retractor muscles are associated with each spicule. A sensory accessory piece connects with the tip of the gubernaculum and protrudes from the lower side of the opening of the spicular pouch; it protracts and retracts with the muscularized gubernaculum. The gubernaculum varies from bow-shaped in the distal part to boat-shaped in the mid region. A sac exits beneath the accessory piece as a buffer for its movement. A cuticular guiding bar originating from the dorsal wall of the spicular pouch has a tongue. The ventral surface of the tongue is sclerotized to separate the two spicules. It is mobile by muscles of the protractor gubernaculi, retractor gubernaculi, and seductor gubernaculi. PMID- 19294123 TI - Evaluation of Morphological Variability in Meloidogyne arenaria. AB - Seven populations, representing cytological race A (triploid, 3n = 51-56) and the two host races (infective and noninfective on peanut) of Meloidogyne arenaria were studied with light microscopy (LM) and scanning electron microscopy (SEM). Characteristics of root-knot nematodes, recently recommended as useful taxonomic traits, were reexamined among these populations, and their variability both within and between populations was ascertained. We found that stylet morphology of females and head and stylet morphologies of males and second-stage juveniles were the most reliable characters for identification. The two host races of M. arenaria could not be distinguished morphologically. Two of the populations could be separated consistently from the remainder but were not sufficiently divergent to be considered new species. These two variant populations were similar; neither produced males in culture, and they differed from the typical populations in female perineal patterns (LM) as well as in cephalic structure (SEM) and tail shape (LM) of second-stage juveniles. In morphometric studies, most characters of the variant populations differed significantly from those of the typical M. arenaria. PMID- 19294124 TI - Behavior of Tethered Meloidogyne incognita. AB - The tethered-nematode technique was adapted for use with second-stage juveniles of Meloidogyne incognita. The data demonstrate that M. incognita exhibits the same patterns of behavior as adults of the free-living nematode, Caenorhabditis elegans. The principal differences are that M. incognita is slower and less regular in its behavior than C. elegans. The frequency of normal waves is about 0.2 Hz; that of reversal waves is about 0.06 Hz. Reversal bouts last about 1 minute. In response to a change in NaCl concentration, M. incognita modulates the probability of initiating a reversal bout in the same manner as C. elegans except that it responds more slowly and is repelled instead of attracted. PMID- 19294125 TI - Influence of Selected Cultural Practices on Winter Survival of Pratylenchus brachyurus and Subsequent Effects on Soybean Yield. AB - Planting date of soybean, Glycine max, influenced winter survival of Pratylenchus brachyurus in microplots at two locations in North Carolina. Delayed planting resulted in a linear decrease (P = 0.05) in the numbers of P. brachyurus at soybean harvest. Effects of planting date on nematode numbers persisted over winter, indicating that survival in the absence of a host is density independent. Compared with winter fallow, winter wheat, Triticum aestivum, reduced winter survival of P. brachyurus. Subsequent soybean yields were suppressed by the overwintering population of this nematode at one location but not at another. PMID- 19294126 TI - Glomus fasciculatum, a Weak Pathogen of Heterodera glycines. AB - The occurrence ofchlamydospores of Glomus fasciculatum (Gf) within cysts of the soybean cyst nematode, Heterodera glycines, and the effects of vesicular arbuscular mycorrhizae on nematode population dynamics and soybean (Glycine max) plant growth were investigated. Chlamydospores occupied 1-24% of cysts recovered from field soil samples. Hyphae of Missouri isolate Gfl penetrated the female nematode cuticle shortly after she ruptured the root epidermis. Convoluted hyphae filled infected eggs, and sporogenesis occurred within infected eggs. G. microcarpum, G. mosseae, and two isolates of Gf were inoculated with H. glycines on plants of 'Essex' soybeans. Each of the two Gf isolates infected about 1% of the nematode eggs in experimental pot cuhures. The Gfl isolate decreased the number of first-generation adult females 26%, compared with the nonmycorrhizal control. The total numbers of first-generation plus second-generation adult females were similar for both Gf isolates and 29-41% greater than the nonmycorrhizal control. Soybean plants with Gf and H. glycines produced more biomass than did nonmycorrhizal plants with nematodes, but only Gfl delayed leaf senescence. PMID- 19294127 TI - Growth and Virulence of Steinernema glaseri Influenced by Different Subspecies of Xenorhabdus nematophilus. AB - Three Xenorhabdus nematophilus subspecies influenced Steinernema glaseri growth profiles and growth rates, but this was not necessarily because of different bacterial growth rates. Virulence of dauer nematodes in larval Galleria mellonella varied with the number of dauers retaining bacteria and the bacterial subspecies. Virulence was least for dauers grown on X. nematophilus subsp. bovienii because of the lack of retained bacteria. Virulence was subsequently restored by culturing these nematodes on X. nematophilus subsp. poinari. PMID- 19294128 TI - Isolation of Subcellular Granules from Second-Stage Juveniles of Meloidogyne incognita. AB - Subcellular granules from the second-stage (preparasitic) juveniles of root-knot nematode Meloidogyne incognita were isolated by isopycnic centrifugation on Percoll. The granules had an apparent density of 1.13 g/cm(3). The relative specific activity of acid phosphatase in the granule extract was 8.4. Acid phosphatase activity was also detected histochemically in the subventral gland granules. Electron microscopy and malate dehydrogenase activity indicated that contamination of granules by mitochondria was negligible. Electrophoresis of the granule extract in the presence of sodium dodecyl sulfate showed 15-20 major protein bands. PMID- 19294129 TI - Applications of biotechnology to nematology: symposium introduction. PMID- 19294130 TI - Chemical communicators in nematodes. AB - Chemical signals released by one organism and perceived by another organism are classified as semiochemicals. Semiochemicals are divided into pheromones, which elicit intraspecific responses, and allelochemics, which elicit interspecific responses. Nematodes utilize and (or) recognize signals from both categories of semiochemicals. The existence of pheromones, specifically sex and aggregation pheromones, has been demonstrated in numerous plant and animal parasitic and free living nematodes. Sex pheromones have been isolated and purified from Nippostrongylus brasiliensis and Heterodera glycines, and epidietic pheromones have been shown to be responsible for initiation of dauer juvenile formation in Caenorhabditis elegans. Allelochemics cause interspecific responses in insects and other invertebrates but are only postulated to occur in nematodes. Food finding behavior of nematodes is almost certainly caused by host-released allelochemic messengers. Understanding of the behavioral responses and the chemical messengers that affect bioregulation of various processes in nematodes will influence future management strategies. PMID- 19294131 TI - Recent developments in nematode steroid biochemistry. AB - Current knowledge of steroid nutrition, metabolism, and function in free-living, plant-parasitic and animal-parasitic nematodes is reviewed, with emphasis upon recent investigation of Caenorhabditis elegans. A number of 4-desmethylsterols with a trans-A/B ring configuration can satisfy the steroid nutritional requirement in C. elegans, but sterols with a cis-A/B ring configuration or trans A/B sterols with a 4-methyl group cannot. C. elegans removes methyl or ethyl substituents at C-24 of the plant sterols sitosterol, campesterol, stigmasterol, stigmastanol, and 24-methylene-cholesterol to produce various sterols with structures partially dependent upon that of the dietary sterol. Additional metabolic steps in C. elegans include reduction of Delta(2)(2)- and Delta-bonds, C-7 dehydrogenation, isomerization of a Delta-bond to a Delta(1)-bond, and 4alpha methylation. An azasteroid and several long-chain alkyl amines interfere with the dealkylation pathway in C. elegans by inhibiting the Delta(2)-sterol reductase; these compounds also inhibit growth and reproduction in various plant-parasitic and animal-parasitic nematodes. A possible hormonal role for various steroids identified in nematodes is discussed. PMID- 19294132 TI - Germplasm modification and its potential for finding new sources of resistance to diseases. AB - In vitro procedures are playing a major role in plant breeding. Embryo rescue, either through the culture of excised embryos derived from incompatible crosses or by means of ovule culture, has been a standard procedure for the introgression of genes conferring disease resistance into economically important plants. Somatic hybridization (i.e., protoplast fusion) has also been demonstrated to have some potential in obtaining hybrids that result from very wide interspecific and intergeneric crosses. Wide crosses have also been achieved by means of in vitro pollination of excised ovaries or ovules. Tissue culture-induced variability in regenerated plant (i.e., somaclonal variation) appears to be an effective way of obtaining undirected genetic change that can enhance disease resistance and yield and alter the growth habit of crops that are normally propagated vegetatively (e.g., potato) or by seed (e.g., tomato). In the near future, the isolation and sequencing of genes that confer resistance to specific plant pathogens will be possible, and transfer of this information between species will become a reality. PMID- 19294133 TI - Characterization and Localization of Saccharides on the Head Region of Four Populations of the Potato Cyst Nematode Globodera rostochiensis and G. pallida. AB - N-acetylglucosamine, galactose, N-acetylgalactosamine and mannose and (or) glucose were present on specimens of two populations of Globodera rostochiensis and two of G. pallida representing four different pathotypes. Individuals within the pathotypes varied in the amounts of some of the saccharides present. The Pa population differed from the other populations in the presence on all individuals of N-acetylgalactosamine and the absence of extensive residues of mannose/glucose. TEM studies showed that N-acetylglucosamine and mannose/glucose were present on the exudate from the amphids of juveniles from the Ro population. PMID- 19294134 TI - Plant parasitic nematodes associated with leatherleaf fern. AB - Seven species of plant parasitic nematodes were found to be associated with leatherleaf fern (Rumohra adiantiformis) in central Florida. Of these, Pratylenchus penetrans, Tylenchorhynchus claytoni, and Criconemoides curvatum were commonly encountered. Nematode communities generally included two or three species of plant parasitic nematodes, with greatest diversity in nematode species occurring in ferneries shaded by oak trees. Species diversity was not correlated with fernery age. Leatherleaf fern was tolerant of P. penetrans and T. claytoni in microplot tests. PMID- 19294135 TI - Variation in Radopholus citrophilus Population Densities in the Citrus Rootstock Carrizo Citrange. AB - Seedlings of the hybrid citrus rootstock, Carrizo citrange (Citrus sinensis x Poncirus trifoliata) do not uniformly limit development of the citrus burrowing nematode, Radopholus citrophilus. Variation in nematode population densities in roots of seedlings germinating from the same seed suggests that factors responsible for nematode incompatibility are not functional or are not inherited uniformly among progeny. Seeds which produced a single seedling were more likely to produce plants which suppressed citrus burrowing nematode population increase than were seeds which produced two or three seedlings. PMID- 19294136 TI - Survival and Horizontal Movement of Infective Stage Neoaplectana carpocapsae in the Field. AB - Infective stage Neoaplectana carpocapsae were applied to a furrow of loam soil in the field. At regular intervals over a 7-week period, soil samples were taken at distances of 0, 10, 20, 30, and 40 cm on either side of the furrow. Live, infective nematodes were recovered from the soil at all sampling dates over the 7 week period, and nematodes slowly spread out from the original line of application, averaging a distance of 4.35 cm/day. PMID- 19294137 TI - Effects of Hydroxyurea on the Ultrastructure of Giant Cells in Galls Induced by Meloidogyne javanica. AB - Hydroxyurea (HU) at concentrations of 10 or 20 mg/liter was included in a medium on which excised tomato roots infected with the root-knot nematode Meloidogyne javanica were grown. In the HU, treated roots, giant cells were small and contained large vacuoles. Giant cell nuclei were amoeboidal with relatively small nucleoli in treated roots, compared with giant cells of nontreated galls. In treated-root giant cells, the cytoplasm was diffuse and few organelles such as mitochondria, dictyosomes, and endoplasmic reticulum were detected; also, walls of giant cells were thin with less extensive ingrowths than in nontreated roots. We conclude that HU suppressed normal giant cell formation interfering with its function as a feeding cell. PMID- 19294138 TI - Pathogenicity of Bursaphelenchus xylophilus on Pines in Minnesota and Wisconsin. AB - The pinewood nematode, Bursaphelenchus xylophilus, was inoculated into established native jack and red pines (Pinus banksiana and P. resinosa) and exotic Austrian pine (P. nigra) in Minnesota and Wisconsin forests during summer 1981. The nematode isolates did not kill established nonstressed pine trees growing in the forest. However, the same nematode isolates killed pine seedlings under greenhouse conditions. Girdling the main stem of some trees to induce stress resulted in the death of the majority of inoculated and noninoculated branches of Austrian and jack pines, but no branch death was observed on red pine. Greater numbers of nematodes were extracted from branches of inoculated, girdled trees than from nongirdled trees. The mean number of nematodes extracted from branches of inoculated, nongirdled trees was 0.3 - 14 nematodes per gram of wood. PMID- 19294139 TI - Characterization of a New Burrowing Nematode Population, Radopholus citrophilus, from Hawaii. AB - Karyotype, host preference, isozyzme patterns, morphometrics, and mating behavior of two burrowing nematode populations from Hawaii, one infecting Anthurium sp. and the second infecting Musa sp., were compared with Radopholus similis and R. citrophilus populations from Florida. The population from Anthurium sp. had five chromosomes (n = 5), and that from Musa sp. had four (n = 4). Neither of the Hawaiian nematode populations persisted in roots of Citrus limon or C. aurantium. Anthurium clarinerivum and A. hookeri were hosts of the burrowing nematode population from anthurium in Hawaii and of R. citrophilus from Florida, whereas the two anthurium species were poor hosts of the population from Musa sp. in Hawaii and R. similis from Florida. The isozyme pattern of the population isolated from anthurium was identical to that of R. citrophigus, whereas the pattern of the population from banana in Hawaii was identical to that of R. similis. Mating behavior between the burrowing nematode population isolated from Anthurium sp. and a Florida population of R. citrophilus supports their close taxonomic relationship. Mating was observed between the population from Anthurium sp. and the Florida population of R. citrophilus but not between the Hawaiian burrowing nematode population isolated from Musa sp. and a Florida population of R. citrophilus. These findings indicate that a previously unidentified population of R. citrophilus which does not parasitize citrus occurs in Hawaii. PMID- 19294140 TI - Reaction of Six Solanum tuberosum Cultivars to Pratylenchus penetrans. AB - Six potato cultivars - Superior, Yukon Gold, Monona, Norchip, Kennebec, and Russet Burbank - were grown in Tioga loamy sand in tile microplots noninfested or infested with 9,800-11,500 P. penetrans/kg soil. At midseason, soil nematode population densities on Superior potatoes were 39,800/kg compared with 17,500 25,800/kg on the other cultivars. At harvest, 118 or 139 days after planting depending on maturity date, the final soil P. penetrans population density was 24,400/kg with Superior and 34,100-51,500/kg with the other cultivars. No differences occurred in the rate of nematode build-up in soil on the six potato cultivars. The nematode suppressed yield of marketable tubers of Superior by 73% and of Yukon Gold by 25%. Losses for Russet Burbank (61%), Kennebec (55%), Monona (46%), and Norchip (43%) were intermediate. PMID- 19294141 TI - Influence of Meloidogyne incognita on Resistant and Susceptible Sweet Potato Cultivars. AB - Effects of several population densities ofMeloidogyne incognita on the sweet potato cultivars Centennial (susceptible) and Jasper (moderately resistant) were studied. Field plots were infested with initial levels (Pi) of 0, 10, 100, 1,000, 5,000, and 10,000 eggs and juveniles/500 cm(3) soil in 1980 and 0, 100, 1,000, 2,000, 3,000, 4,000, and 5,000 in 1981. M. incognita population development trends were similar on both cultivars; however, at high Pi, more eggs and juveniles were recovered from Centennial than from Jasper. The highest Pi did not result in the highest mid-season (Pm) counts. Pi was negatively correlated with the number of marketable roots and root weight but positively correlated with total cracked roots, percentage of cracked roots, and cracking severity. Jasper tolerated higher Pi with greater yields and better root quality than Centennial. Cracking of fleshy roots occurred with both cultivars at low Pi. PMID- 19294142 TI - Control of Ditylenchus dipsaci in Infected Garlic Seed Cloves by Nonfumigant Nematicides. AB - Different rates of granular formulations ofaldicarb, carbofuran, ethoprop, fensulfothion, and phenamiphos were applied directly onto garlic seed cloves in the seed furrow in sandy clay loam, clay loam, and loam soils at planting to assess efficacy for control of Ditylenchus dipsaci in infected seed cloves. All treatments were compared to hotwater-formalin clove dip disinfection treatment and to nontreated infected controls. Aldicarb and phenamiphos at 2.52 and 5.04 kg a.i./ ha, but not at lower rates, effectively suppressed infection by D. dipsaci and increased yields. Although both nematicides slightly slowed the rate of plant emergence, normal stands were established. Trace levels of infection occurred in all treatments, including the hotwater-formalin dip. Carbofuran at 5.04 kg a.i./ha controlled the nematode but was phytotoxic. Ethoprop was phytotoxic. Fensulfothion did not control D. dipsaci even at the highest application rate, 8.90 kg a.i./ha. Single and multiple applications of oxamyl at 1.12-8.96 kg a.i./ha, applied as a surface spray or in furrow irrigation water, slowed the early progression of disease symptoms but failed to provide season-long nematode control. PMID- 19294143 TI - Management of Globodera rostochiensis as Influenced by Nematode Population Densities and Soil Type. AB - The effects of aldicarb, oxamyl, 1,3-D, and plastic mulch (solarization) on soil population densities of the golden nematode (GN) Globodera rostochiensis was assessed in field and microplot experiments with different soil types. Oxamyl was evaluated in both soil and foliar treatments, whereas aldicarb, 1,3-D, and solarization were applied only to soil. Soil applications of aldicarb and oxamyl resulted in reduced nematode populations after GN-susceptible potatoes in plots with initial population densities (Pi) of > 20 and 7.5 eggs/cm(3) soil, respectively, but nematode populations increased in treated soil when Pi were less than 20 and 7.5 eggs/cm(3)soil. In clay loam field plots with Pi of 19-76 eggs/cm(3) soil, nematode densities increased even with repeated foliar applications of oxamyl, whereas nematode populations at Pi greater than 76 eggs/cm(3) soil were reduced by foliar oxamyl. Treatment with 1,3-D or solarization, singly or in combination, reduced GN soil population densities regardless of soil type or Pi. Temperatures lethal to GN were achieved 5 cm deep under clear plastic but not 10 or 15 cm deep. PMID- 19294144 TI - Biology and Influence of Pterotylenchus cecidogenus on Desmodium ovalifolium. AB - Pterotylenchus cecidogenus did not inhibit germination of Desmodium ovalifolium but did reduce survival of seedlings and root and stem growth. Stem gall formation was associated with nematode population densities which were positively correlated with plant age at time of inoculation. Nematode population densities increased 100 times in 52 days following inoculation. PMID- 19294145 TI - Genotypic differentiation of meloidogyne populations by detection of restriction fragment length difference in total DNA. AB - Detection of EcoRI restriction fragment length differences in repetitive DNA sequences permitted the rapid diagnosis, by genotype, of randomly selected populations of Meloidogyne incognita, Races 1, 2, 3, and 4; M. javanica; M. arenaria, Races 1 and 2; and M. hapla, Races A and B. PMID- 19294146 TI - Parasitism of Xiphinema rivesi and X. americanum by Zoosporic Fungi. AB - Living Xiphinema americanum (Xa) and X. rivesi (Xr) extracted from soil samples and stored for 1-5 days at 4 or 20 C contained aseptate fungal hyphae. The fungi directly penetrated the nematode's cuticle from spores encysted near the head. Penetration through the stoma, vulva, or anus was rare. Catenaria anguillulae (Cat), Lagenidium caudatura (Lag), Aphanomyces sp. (Aph), and Leptolegnia sp. (Lep) were isolated into pure culture from infected nematodes. The pathogenicity of these zoosporic fungi was determined by incubating mixed freshly extracted Xa and Xr in 2% soil extract (pH = 6.7, conductivity = 48 mumhos, 20 +/- 2 C) containing zoospores obtained from single-spore isolates. After 4 days, Cat, Lag, Aph, and Lep had infected 78, 18, 13, and 22%, respectively, of the nematodes. Both Xa and Xr were infected by every fungus; however, the relative susceptibility of Xa and Xr to these fungi was not determined. All noninoculated control nematodes remained uninfected and alive. In a second experiment, parasitism of Xa and Xr by Aph and Lep was increased when nematodes were incubated in 2% soil extract for 4 days before exposure to zoospores. In a third experiment, parasitism of Xa and Xr by Cat was greater in diluted saturation soil extract (conductivity = 100-400 mumhos) than in undiluted saturation extract (conductivity = 780 mumhos). Cat produced small zoospores (4-mum-d), bulbous infection hyphae, and assimilative hyphae of varying diameters in nematodes, whereas Lag, Aph, and Lep produced large zoospores (8-mum-d) and tubular, uniform infection and assimilative hyphae in nematodes. PMID- 19294147 TI - Susceptibility of Psorophora columbiae Larvae over Time to Parasitism by Romanomermis culicivorax. AB - The ability of Romanomermis culicivorax preparasites to penetrate and infect Psorophora columbiae decreased substantially after ca. 28 hours. Parasitism at temperatures typical of Louisiana rice fields (i.e., 26, 29, and 32 +/- 0.5 C) showed a significant linear decrease (P < 0.01) as the percentage of older larval instars increased at the times of exposure. These data emphasize the need for a synchronous field application of preparasites to challenge the rapid development of early instars of Ps. columbiae. Applications of postparasites rather than insecticide treatments to potential mosquito breeding habitats may offer greater flexibility in larval mosquito control programs. PMID- 19294148 TI - Field Interrelationships among Heterodera glycines, Pratylenchus scribneri, and Three Other Nematode Species Associated with Soybean. AB - Field experiments were conducted in 1982 and 1983 to assess interactions between Heterodera glycines and Pratylenchus scribneri on soybean in southern Illinois. Soybean cyst nematode susceptible cultivar Williams 79 and resistant cultivar Fayette were treated or not treated with aldicarb 15G. Initial population densities were 35 H. glycines cysts containing eggs, 100 P. scribneri, 30 Helicotylenchus pseudorobustus, 225 Paratylenchus projectus, and 85 Tylenchorhynchus martini per 250 cm(3) soil in 1982, whereas in 1983 populations were 11 H. glycines cysts, 330 P. scribneri, and 620 H. pseudorobustus. In both years H. glycines populations increased on nontreated Williams 79, decreased on both treated and nontreated Fayette, and remained at initial levels on treated Williams 79. Recovery of P. scribneri per gram dry root was different between nontreated cultivars in 1982 but not in 1983. Aldicarb treatment suppressed soil and root populations of P. scribneri on both cultivars in both years. Populations of H. pseudorobustus, P. projectus, and T. martini at harvest indicated little population increase on either nontreated cultivar. In 1982 H. glycines caused yield suppression but P. scribneri did not, as differences in yield occurred between cultivars but not between aldicarb treatments. In 1983, however, there were no yield differences between cultivars, but aldicarb application resulted in yield increase in both cultivars. In 1983 the yield increase resulting from P. scribneri control was approximately 25%. No synergistic effect on yield was observed between H. glycines and P. scribneri. PMID- 19294149 TI - Interrelationships of Meloidogyne arenaria and M. incognita on Tolerant Soybean. AB - Reproduction of Meloidogyne arenaria race 2 was excellent on Centennial, Govan, and Kirby soybeans, the latter two of which have tolerance to this species. The M. incognita race 1 isolate reproduced poorly on Centennial, especially at the higher of two temperature regimes. Numbers of galls and egg masses of M. arenaria plus M. incognita in simultaneous equivalent infestations on Centennial did not differ from sequential infestations in which M. arenaria was added first and M. incognita was added to the same pots, 1,2, or 3 weeks later. However, at both 25 and 30 C, suppression of galls and egg masses occurred when inoculation of M. incognita preceded that of M. arenaria by 2 weeks. Generally, M. arenaria reproduced well at 25 or 30 C, whereas M. incognita reproduced better at 30 C. Kirby was tolerant to either nematode species at 25 and 30 C, but in combined infestations of M. arenaria and M. incognita there was evidence of synergistic growth suppression. Govan was tolerant of M. arenaria at 25 C but not at 30 C. Moreover, general plant growth was less vigorous for Govan at the higher temperature, whereas Centennial was much more vigorous at this temperature. Kirby grew equally well at both temperatures. PMID- 19294150 TI - Host Range of, and Plant Reaction to, Subanguina picridis. AB - The host range of the knapweed nematode, Subanguina picridis (Kirjanova) Brzeski, under controlled environmental conditions was extended to include, in addition to Russian knapweed, Acroptilon repens (L.) DC., plant species within the Centaureinae, and Carduinae subtribes of the Cynareae tribe of the Asteraceae family. Examination of host response to nematode infection revealed that Russian knapweed was the only highly susceptible host plant. Diffuse knapweed (Centaurea diffusa Lam.) was moderately susceptible, and other plants which formed galls were resistant to S. picridis. PMID- 19294152 TI - Modification of proprietary chemicals for increasing efficacy. AB - Several factors limit the efficacy of nematicides. Consequently, modifications can be made to their active ingredient, formulation, and application to increase their efficacy. PMID- 19294151 TI - New trends in managing plant parasitic nematodes-introductory remarks. PMID- 19294153 TI - Organic and inorganic nitrogen amendments to soil as nematode suppressants. AB - Inorganic fertilizers containing ammoniacal nitrogen or formulations releasing this form of N in the soil are most effective for suppressing nematode populations. Anhydrous ammonia has been shown to reduce soil populations of Tylenchorhynchus claytoni, Helicotylenchus dihystera, and Heterodera glycines. The rates required to obtain significant suppression of nematode populations are generally in excess of 150 kg N/ha. Urea also suppresses several nematode species, including Meloidogyne spp., when applied at rates above 300 kg N/ha. Additional available carbon must be provided with urea to permit soil microorganisms to metabolize excess N and avoid phytotoxic effects. There is a direct relation between the amount of "protein" N in organic amendments and their effectiveness as nematode population suppressants. Most nematicidal amendments are oil cakes, or animal excrements containing 2-7% (w:w) N; these materials are effective at rates of 4-10 t/ha. Organic soil amendments containing mucopolysaccharides (e.g., mycelial wastes, chitinous matter) are also effective nematode suppressants. PMID- 19294154 TI - Impact of conservation tillage on nematode populations. AB - Literature reporting the development of conservation tillage and the research that has been conducted on nematode control in crops grown in conservation tillage systems is reviewed. Effects of different types of conservation tillage on population densities of various nematode species in monocropping and multicropping systems, effects of tillage on nematode distribution in the soil profile, effects of conservation tillage on nematode control, and the role of nematology in conservation tillage research are discussed. PMID- 19294155 TI - Interaction of Vesicular-Arbuscular Mycorrhizae and Cultivars of Alfalfa Susceptible and Resistant to Meloidogyne hapla. AB - The interaction between vesicular-arbuscular mycorrhizal (VAM) fungi and the root knot nematode (Meloidogyne hapla) was investigated using both nematode susceptible (Grasslands Wairau) and nematode-resistant (Nevada Synthetic XX) cultivars of alfalfa (Medicago sativa) at four levels of applied phosphate. Mycorrhizal inoculation improved plant growth and reduced nematode numbers and adult development in roots in dually infected cultures of the susceptible cultivar. The tolerance of plants to nematode infection and development when preinfected with mycorrhizal fungi was no greater than when they were inoculated with nematodes and mycorrhizal fungi simultaneously. Growth of plants of the resistant cultivar was unaffected by nematode inoculation but was improved by mycorrhizal inoculation. Numbers of nematode juveniles were lower in the roots of the resistant than of the susceptible cultivar and were further reduced by mycorrhizal inoculation, although no adult nematodes developed in any resistant cultivar treatment. Inoculation of alfalfa with VAM fungi increased the tolerance and resistance of a cultivar susceptible to M. hapla and improved the resistance of a resistant cultivar. PMID- 19294156 TI - Biology of Subanguina picridis, a Potential Biological Control Agent of Russian Knapweed. AB - The knapweed nematode, Subanguina picridis, forms galls on the leaves, stems, and root collar of Russian knapweed, Acroptilon repens. After being revived from a dormant, cryptobiotic state, second-stage juveniles required at least 1 month in a free-living state before becoming infective. Galls were induced on relatively slow-growing host plants that retained their apical meristems at or near the soil surface for 2-5 weeks. Galls developed extensive areas of nutritive tissue. The nematode was introduced from the Soviet Union and released in Canada for the biological control of Russian knapweed. PMID- 19294157 TI - Morphological and Biological Parameters of the Knapweed Nematode, Subanguina picridis. AB - Specimens of the knapweed nematode Subanguina picridis (Kirjanova) Brzeski obtained from different host plants were highly variable in measurement and structure. This variability refutes the validity of six Subanguina species attacking plants in the Asteraceae. PMID- 19294158 TI - Effects of Initial Nematode Density on Population Dynamics of Globodera rostochiensis on Resistant and Susceptible Potatoes. AB - The influence of resistant and susceptible potato cultivars on Globodera rostochiensis population density changes was studied at different nematode inoculum levels (Pi) in the greenhouse and field. Soil in which one susceptible and two resistant cultivars were grown and fallow soil in pots was infested with cysts to result in densities of 0.04-75 eggs/cm(3) soil. A resistant cultivar was grown in an infested field with Pi of 0.7-16.7 eggs/cm(3) soil. Pi was positively correlated with decline of soil population densities due to hatch where resistant potatoes were grown in the greenhouse and in the field but not in fallow soil. However, Pi was not correlated with in vitro hatch of G. rostochiensis cysts in water or potato root diffusate. Under continuous culture o f a resistant cultivar, viable eggs per cyst declined 60-90% per plant growth cycle (4 weeks) and the number of cysts containing viable eggs had decreased by 77% after five cycles. The rate of G. rostochiensis reproduction on both resistant and susceptible cultivars was negatively correlated with Pi. These data were used to predict the effect of resistant and susceptible potato cultivars on G. rostochiensis soil population dynamics. PMID- 19294159 TI - Globodera rostochiensis Population Density Decline in Relation to Spatial Distribution around Resistant Potato Plants. AB - Golden nematode, Globodera rostochiensis (GN) population decline under resistant potatoes was related to cyst distance from plants 23 cm apart in rows 92 cm apart. GN decline, determined by sampling an infested field planted to the resistant cultivar 'Yankee Chipper', was 81.8% in cores 11.5 cm from plants within rows. Decline was 27.4% at 23 cm from plants between rows and 36.6% at 46 cm. Population decline of juveniles in cysts added to soil in bags was 90.3% for cysts 11.5 cm from plants within rows planted to the resistant cultivar 'Rosa'. Decline between rows was 83.5, 76.9, and 60.4% at 11.5, 23.0, and 46.0 cm from plants, respectively. Maximum decline within for rows 30.5, 46.0, 61.0, and 92.0 cm apart, respectively. Decline under fallow was 43.5%, signif- which peaked 7 weeks AE. There was no effect of soil depth on population decline at any sampling position. Decreasing row spacing resulted in 79.9, 74.2, 73.4, and 66.1% GN population decline for rows 30.5, 46.0, 61.0, and 92.0 cm apart, respectively. Decline under fallow was 43.5%, significantly less than under potatoes. Potato root weight between rows was negatively correlated with row spacing and positively correlated with GN population decline. PMID- 19294160 TI - Heterodera glycines in Indiana: I. Reproduction of Geographical Isolates on Soybean Differentials. AB - Four of five geographical isolates of Heterodera glycines from Indiana classified as Race 3 using standard differentials showed many differences when classified using another group of differentials comprised of five soybean breeding lines and cultivars. Two isolates from northern Indiana produced cysts on more of the differentials tested than did three isolates from southern Indiana, suggesting that potential resistant lines should be tested on a range of H. glycines populations originating from the areas for which cultivars are being developed. PMID- 19294161 TI - Heterodera glycines in Indiana: II. Morphology of Geographical Isolates. AB - Although much morphometric overlap occurs among five geographical isolates of Heterodera glycines in Indiana, significant differences in means exist among the isolates for various comparisons of second-stage juveniles. By using combinations of means, most of the isolates can be distinguished from the rest: e.g., the Vanderburgh County isolate (southern Indiana) has the longest esophagus, tail, and tail terminus; the Vigo County isolate (also from the south) has the shortest esophagus; the White County isolate (northern Indiana) has the shortest tail and tail terminus and the greatest total length; the Benton County isolate (north) is the shortest. Morphological similarities and differences do not appear to be coordinated with reproductive behavioral patterns we observed in the northern versus the southern isolates. PMID- 19294162 TI - Heterodera glycines in Indiana: III. 2-D Protein Patterns of Geographical Isolates. AB - Protein patterns obtained by two-dimensional polyacrylamide gel electrophoresis for three isolates of Heterodera glycines from southern Indiana appear qualitatively similar and have higher pairwise Jaccard similarity coefficients with each other than with isolates from northern Indiana. Three isolates from three northern counties share proteins not present in the southern isolates, but as a group the northern isolates are less similar to each other than are the southern Indiana isolates. PMID- 19294163 TI - Spatial Analysis of Heterodera glycines Populations in Field Plots. AB - Spatial heterogeneity in nematode population densities presents an obstacle to the precise determination of infestation levels. Three field plots were intensively sampled for soybean cyst nematode (Heterodera glycines Ich.) cysts before and after spring cultivation to quantify the spatial attributes of the population. Population density strata were detected running parallel to plant rows. Highest population densities before cultivation were found in the plant row and the middle furrow, Population density in the plant row averaged 26% higher and 4% lower than the whole-plot mean before and after cultivation, respectively. Cysts containing fewer than 25 eggs were not stratified, indicating that most were produced before the previous season. Sample population counts were fit to the negative binomial distribution model before cultivation, but distributions differed among plots. The Neyman type A and negative binomial distributions both fit the data after cultivation disturbed the soil. Population clusters 1-3 m long were detected in plant beds before cultivation. Heterogeneity in population density increased with plant row length after cultivation. Optimum plot length for minimal spatial heterogeneity in four-row mechanically tended field plots was estimated at 6 m after trimming plot ends. PMID- 19294164 TI - Improving the accuracy of sampling field plots for plant-parasitic nematodes. AB - The validity of nematode data from field experiments depends largely on how well samples represent the nematode population. Data from an intensive sampling of three field plots before and after spring cultivation were used to compare eight simulated sampling schemes. Average deviation from the plot mean ranged from 10% to 34% before cultivation and from 7% to 16% after cultivation. Samples taken from only the plant row erred most before cultivation but were comparable to other schemes after cultivation. Several schemes achieved a 25% deviation or less in 90% of the sample simulations. Sampling a nematode population usually involves subsampling a composite bulk sample, however, and this increases error by an estimable amount. A random sample with 35 cores and four random subsamples estimated mean plot densities within 25% with probabilities ranging from 0.77 to 0.85. The probability of a sample-subsample combination coming within a specified percent error of the true mean can be extended cautiously to any field mean and variance more-or-less independent of species and area using formulae presented herein. The most economical method of increasing sample accuracy was to increase the number of soil cores. PMID- 19294165 TI - Tolerance of Soybean to Heterodera glycines. AB - Seven soybeans were selected from 200 entries evaluated for tolerance to soybean cyst nematode (SCN), Heterodera glycines. Tolerance to SCN was measured by comparing the seed yield from aldicarb-treated vs. nontreated plots. A yield response index (YRI) was calculated for each entry: YRI = (seed yield from nontreated plot/seed yield from treated plot) x 100. The soybean entries Coker 156, PI 97100, and S79-8059 exhibited high tolerance (YRI) to SCN when compared to Essex even though they became heavily infected with SCN. Tolerance in soybeans to SCN may be useful in pest management programs designed to stabilize soybean yield. PMID- 19294166 TI - Inbreeding and hybridizing cyst nematodes on pruned soybeans in petri plates. AB - Inbred nematodes propagated on a selecting host are likely to have homozygous genes of interest for investigating the genetics of host-parasite associations. A technique is presented to inbreed soybean cyst nematodes, by sibling matings at each generation, and to cross inbred lines. Soybean seedlings with severely trimmed cotyledons survive well on 0.8% agar. Eggs from a single female are incubated in water in a microtiter well. Virgin as well as mated females result from inoculation of two juveniles per root. Sibling males from the same source are produced by mass inoculations of eggs. Males are added individually to unmated females. Overall success for fertile females was 14% in 1,368 isolations. Three generations of inbreeding by siblings were achieved using nematodes from two populations that differ in their ability to reproduce on differential soybeans. Hybrids from crosses of the two inbred lines tested on differential hosts showed that the influence of Population 1 (selected and inbred on PI 209332) is greater than that of Population 2 (selected and inbred on PI 89772). Reciprocal crosses suggest that the influence of males is stronger than that of females in determining host specificity of F offspring in these crosses. Our technique is simple and effective for inbreeding and crossing soybean cyst nematodes. PMID- 19294167 TI - Distribution of field corn roots and parasitic nematodes in subsoiled and nonsubsoiled soil. AB - A field trial was conducted for 2 years in an Arredondo fine sand containing a tillage pan at 15-20 cm deep to determine the influence of subsoiling on the distribution of corn roots and plant-parasitic nematodes. Soil samples were taken at various depths and row positions at 30, 60, and 90 days after planting in field corn subsoiled under the row with two chisels and in non-subsoiled corn. At 30 and 60 days, in-row nematode population densities to 60 cm deep were not affected by subsoiling compared with population densities in nonsubsoiled plots. After 90 days, subsoiling had not affected total root length or root weight at the 20 depth-row position sampling combinations, but population densities of Meloidogyne incognita and Criconemella spp. had increased in subsoiled corn. Numbers of Pratylenchus zeae were not affected. Subsoiling generally resulted in a change in distribution of corn roots and nematodes in the soil profile but caused little total increase in either roots or numbers of nematodes. Corn yield was increased by subsoiling. PMID- 19294168 TI - Interaction of Endomycorrhizal Fungi, Superphosphate, and Meloidogyne incognita on Cotton in Microplot and Field Studies. AB - Microplot and field experiments were conducted to determine the effects of two vesicular-arbuscular mycorrhizal (VAM) fungi, Glomus intraradices (Gi) and Gigaspora margarita (Gm), and dicalcium phosphate (P) on Meloidogyne incognita (Mi) reproduction and seed cotton yield of the Mi-susceptible cotton cultivar, Stoneville 213. In 1983 population densities of Mi juveniles were significantly lower 60 and 90 days after planting in microplots receiving Gi. Mycorrhizal fungi reduced the severity of yield losses to Mi, whereas P fertilization increased yield losses to Mi. In 1984 microplot yields were reduced linearly as nematode inoculum densities increased in treatments of Mi alone, Gm, or P, but the response was curvilinear with Gi. Yield suppressions in the 1984 field experiment occurred only in plots infested with Mi alone. In the 1984 microplots, numbers of Mi juveniles penetrating seedling roots increased Iinearly with increasing nematode inoculum densities and was favored when mycorrhizal fungi or superphosphate were added. Juvenile penetration of roots was negatively correlated with yields in all treatments (r = -0.54 to -0.81) except Gm and with number of bolls in Mi alone (r = -0.85) and P (r = -0.81) treatments. Mycorrhizal fungi can increase host tolerance to M. incognita in field conditions and may function as important biological control agents in soils infested with high population densities of efficient VAM species. PMID- 19294169 TI - Association of Criconemella xenoplax and Fusarium spp. with Root Necrosis and Growth of Peach. AB - Criconemella xenoplax, Fusarium solani, and F. oxysporum caused necrosis of Nemaguard peach feeder roots in greenhouse tests. Root necrosis was more extensive in the presence of either fungus than wtih C. xenoplax alone. Shoot growth and plant height were less for plants inoculated with F. oxysporum or F. solani than for plants inoculated with the fungi plus C. xenoplax. Neither synergistic nor additive effects on root necrosis or plant growth occurred between C. xenoplax and the fungal pathogens. PMID- 19294170 TI - Respiration Rate of Steinernema feltiae Infective Juveniles at Several Constant Temperatures. AB - Respiration was measured in dauer stages of the insect-parasitic nematode Steinernema feltiae (= Neoaplectana carpocapsae) at 7, 17, and 27 C. Respiration, Q, and nematode viability were temperature dependent. Mean O consumption for 5 x 10 nematodes the first 24 hours was 0.27 ml at 7 C, 0.83 ml at 17 C, and 2.68 ml at 27 C. The Q was 3.10 for 7-17 C and 3.24 for 17-27 C. Some nematodes died during 2, 14, and 21 days at 27, 17, and 7 C, respectively. The respiratory quotient was below 1 at all temperatures tested. A standard asymptotic model is expressed as oxygen consumed = 2.77 * {1 - exponent[-time * exponent(-B + C * temperature)]}; where 2.77 is the maximum response at 27 C. This model estimates nematode O consumption and viability at storage temperatures between 7 and 27 C. The nematodes died when the O concentration reached 0.5 ml/5 x 10 nematodes. This model may be used to predict O requirements of S. feltiae infective juveniles when stored as a waterless concentrate. PMID- 19294171 TI - Susceptibility of Neoaplectana spp. and Heterorhabditis heliothidis to the Endoparasitic Fungus Drechmeria coniospora. AB - Adhesive conidia of the nematophagous fungus, Drechmeria coniospora (Drechsler) W. Gams and Jansson (Moniliales: Deuteromycetes), would occasionally attach but never penetrate the infective stages of insect parasitic Neoaplectana carpocapsae, N. glaseri, N. bibionis, N. intermedia, and Heterorhabditis helfothidis (Rhabditida). However, adult and pre-infective stages of Neoaplectana spp. became infected by the fungus. PMID- 19294172 TI - Pathotypes of the Pinewood Nematode Bursaphelenchus xylophilus. AB - An isolate of Bursaphelenchus xylophilus from Pinus sylvestris in Missouri infected and reproduced in 2-3-year-old seedlings of P. sylvestris and to some extent in seedlings of P. nigra. Wilting, however, occurred only in P. sylvestris. B. xylophilus isolated from P. strobus in Vermont infected and reproduced only in P. strobus seedlings. P. taeda seedlings were resistant to both of these isolates. Phytotoxin production was seen only in susceptible seedling species-nematode combinations. Significant water loss occurred only in those seedlings that were wilted because of infection by a compatible nematode isolate. Our results suggest that these isolates are pathotypes of B. xylophilus. PMID- 19294173 TI - Morphometrics, Illustration, and Histopathology of Sphaeronema rumicis on Cottonwood in Utah. AB - The morphology of a population of Sphaeronema rumicis Kir'yanova found on cottonwood in Utah is illustrated by light and scanning electron micrographs, as well as by drawings. This is the first report of males of S. rumicis, a species also not known previously to occur in North America. S. rumicis females on cottonwood in the United States were smaller than those found by Kir'yanova on sorrel in the USSR. Females and second-stage juveniles (J2) from the United States had slightly shorter stylets than did females and J2 from the USSR. Males were vermiform and had degenerate esophagi. On secondary cottonwood roots S. rumicis induces formation of a syncytium originating from proliferated pericyclic cells. Thick outer walls, wall protuberances, absence of cell wall ingrowths, dense cytoplasm, and hypertrophied nuclei were the main characteristics of syncytia observed in S. rumicis-infected cottonwood roots. PMID- 19294174 TI - Divisispiculimermis mirus n. gen., n. sp. (Mermithidae: Nematoda) Parasitizing Midges in Cordoba, Argentina. AB - Divisispiculimermis mirus n. gen., n. sp., a mermithid parasitizing larvae of Chironomus sp. in the Cajon o Grande Stream, Cordoba, Argentina, is described. The new genus differs from all other mermithid genera in having paired spicules which are separated and divided into proximal and distal sectors. The other diagnostic characters of the genus are medium size, nematodes with the cuticle appearing smooth (lacking cross fibers under the light microscope); head separated from the rest of the body by a slight constriction at the level of the amphids, six cephalic papillae, mouth papillae absent, mouth opening posterior to level of cephalic papillae; six hypodermal chords at midbody; weakly S-shaped vagina; postparasitic juvenile with a tail appendage. PMID- 19294175 TI - Descriptions of four new species of criconematoidea (tylenchina: nemata) from southern chile. AB - Four new species of Criconematoidea are described from Hoste Island, Chile. Criconema certesi n. sp. is distinguished by the fine, spine-like, cuticular extensions on body annuli; projection of annuli into rows of scales on posterior part of body; single, smooth, labial annulus set off by short collar from second (first body) annulus which is about same diameter as first (labial) annulus. Male with prominent caudal alae, slender curved spicules, and four incisures in lateral field. Ogma terrestris n. sp. is distinguished by small scales with rounded tips bearing minute, short bristles; scales number 21 at mid-body; and first (labial) annulus rounded, not retrorse, not set off from succeeding annuli, narrower in diameter from second (first body) annulus. Hemicycliophora macrodorata n. sp. is distinguished by its large size (L = 1.52 [1.28-1.72] mm); large stylet (146 [127-161] mum); annuli = 297 (280-315); tail slightly spicate, lateral field with or without interruptions of incisures, occasional anastomoses; and males with U-shaped spicules. Paratylenchus fueguensis n. sp. is distinguished by its prominent stylet with large, rounded knobs (4-5 mum across); cephalic region rounded not at all set off; lateral field with four incisures; lateral vulvar membranes present; and male tail short, strongly curved (almost 180 degrees ) ventrad. PMID- 19294176 TI - Histopathology of Root Gall Induced in Tomato by Globodera pallida. PMID- 19294177 TI - Temperature Effects on the Growth and Virulence of Steinernema feltiae Strains and Heterorhabditis heliothidis. PMID- 19294178 TI - Plant parasitic nematodes in new jersey. PMID- 19294179 TI - Comparative Morphology of Atalodera spp. and Thecavermiculatus spp. (Heteroderidae) with Scanning Electron Microscopy. AB - SEM examination of second-stage juveniles (J2) and adults of Atalodera ucri, A. lonicerae (syn. Sherodera lonicerae), Thecavermiculatus sp. (undescribed new species), T. andinus, and T. crassicrustatus revealed new characters. A primitive en face pattern with six separate lips occurs in J2 of Thecavermiculatus spp. examined and in about half the polymorphic A. lonicerae. A derived en face pattern with fused adjacent submedial lips occurs in the other half of A. lonicerae and all A. ucri. Posteriorly, the J2 head of all species is annulated. The primitive en face pattern also occurs in males of A. lonicerae and Thecavermiculatus spp., and posteriorly the head of these species consists of plates. Fewer plates occur rarely in males of A. ucri. Males of A. ucri have a derived en face pattern where lips are fused and the head is annulated. Fusion of lips occurs rarely in males of A. lonicerae. Females of all species have similar derived en face patterns. En face patterns of J2 and males o f Atalodera and Thecavermiculatus may aid in species identification and to elucidate intergeneric relationships, but en face characters shared by the two genera are primitive and are not useful for demonstrating monophyly. Perineal region of females indicates the closeness of the vulval-anal distance, as a derived character, which is shared by Atalodera and most Thecavermiculatus spp. suggesting possible monophyly. T. andinus, while having a similar en face pattern to J2 of other Thecavermiculatus species, lacks the derived character of the perineal region. Phasmid openings were not observed in adults of any of the species examined. PMID- 19294180 TI - Species-Specific Restriction Site Polymorphism in Root-knot Nematode Mitochondrial DNA. AB - Research was initiated to physically characterize the mitochondrial genomes of several Meloidogyne spp. and host-races, to address questions regarding their systematics and dispersal, and to assess the possibility of developing molecular diagnostics for these nematodes. Techniques were developed for purification and rapid detection of mitochondrial DNA from root-knot nematodes. Mitochondrial DNAs among Meloidogyne spp. were demonstrated to exhibit extensive divergence. The potential for using the rapidly diverging mitochondrial genomes as a diagnostic assay for M. incognita, M. hapla, M. arenaria, and M. javanica is discussed. PMID- 19294181 TI - Cuticular Collagenous Proteins of Second-stage Juveniles and Adult Females of Meloidogyne incognita: Isolation and Partial Characterization. AB - Cuticles isolated from second-stage juveniles and adult females of Meloidogyne incognita were purified by treatment with 1% sodium dodecyl sulfate (SDS). The juvenile cuticle was composed of three zones differing in their solubility in beta-mercaptoethanol (BME). Proteins in the cortical and median zones were partially soluble in BME, whereas the basal zone was the least soluble. The BME soluble proteins from the juvenile cuticle were separated into 12 bands by SDS polyacrylamide gel electrophoresis and characterized as collagenous proteins based on their sensitivity to collagenase and amino acid composition. The adult cuticle consisted of two zones which were dissolved extensively by BME. The basal zone was completely solubilized, leaving behind a network of fibers corresponding to the cortical zone. The BME-soluble proteins from the adult cuticle were separated by electrophoresis into nine bands one of which constituted > 55% of the total BME-soluble proteins. All bands were characterized as collagenous proteins. Collagenous proteins from juvenile cuticles also contained glycoproteins which were absent from the adult cuticles. PMID- 19294182 TI - Small Collagenous Proteins Present during the Molt in Caenorhabditis elegans. AB - Immunoblotting experiments using antibodies directed against the large collagenous cuticle proteins of Caenorhabditis elegans revealed a class of small collagenous proteins (CP) of apparent molecular weight 38,000-52,000 present during the L4 to adult molt. These CP are smaller than most vertebrate collagens characterized to date and share many characteristics with the small collagenous products translated in vitro from RNA isolated at this molt. C. elegans collagen genes, collagen-coding mRNA, and collagenous in vitro products that have been characterized are also small. Detection of small CP in vivo in C. elegans thus lends further support to the hypothesis that such small collagenous proteins are the primary gene product precursors to the larger collagenous proteins isolated from the C. elegans cuticle. PMID- 19294183 TI - Steinernema feltiae (DD-136) and S. glaseri: Persistence in Soil and Bark Compost and Their Influence on Native Nematodes. AB - Infective juveniles (J3) of the entomogenous nematodes Steinernema feltiae DD-136 (ca. 10,000 J3/100 ml) and S. glaseri (ca. 2,500 J3/100 ml) were incubated in steam-sterilized and nonsterilized sandy soil and bark compost for 8 weeks at 25 C. The nematodes were recovered by a two-step extraction procedure at 1-week intervals, and their infectivity to lepidopterous larvae (Spodoptera litura and Galleria mellonella) and their effect on the population and community of native nematodes in soil were determined. Survival of inoculated nematodes and mortality of insects were enhanced in sterilized media. Nonsterilized bark compost proved to be equally as suitable a medium as sterilized compost. In nonsterilized soil, the survival curve of S.feltiae declined more rapidly than that or S. glaseri which was less infective to insects despite its greater persistence even in nonsterilized soil. Soon after the addition of steinernematids to soil, the population of native nematodes showed a fluctuation with an increase in rhabditids and a decrease in other kinds of nematodes. PMID- 19294184 TI - Strelkovimermis spiculatus n. sp. (Mermithidae: Nematoda) Parasitizing Aedes albifasciatus Mac. (Culicidae: Diptera) in Argentina. AB - Strelkovimermis spiculatus n. sp. (Mermithidae: Nematoda), a parasite of the mosquito Aedes albifasciatus Mac. in Argentina, is described. Diagnostic characters of this species include a ventrally shifted mouth opening and spicules fused at the tips. The mermithid has been reared on Culex pipiens L. larvae in the laboratory. PMID- 19294185 TI - Effects of Temperature, Plant Age, Soil Texture, and Meloidogyne incognita on Early Growth of Soybean. AB - A digitizer-microcomputer combination was utilized to determine soybean seedling response to population densities of M. incognita (Mi) under varied environmental conditions. Plant age, temperature, soil texture, and initial Mi inoculum (Pi) influenced the pattern of shoot and root growth. Effects of Mi on plant top growth were evident on plants inoculated 2 days after seeding, but generally were not noticeable on those receiving Mi after 4, 6, or 8 days (observations limited to 6 days after inoculation). The greatest Pi of Mi (16,700 juveniles/plant) suppressed root growth on plants inoculated at 2 or 4 days after seeding. Mi had no impact on root growth at 22 C on plants inoculated 6 or 8 days after seeding at any temperature used (22, 26, 30 C). New root initiation was inhibited on soybeans inoculated 2 days after seeding at the highest Pi at all three temperatures, but only at 30 C for a Pi of 1,670 juveniles/plant. Growth of first order lateral roots and general root length were suppressed by Mi on the youngest (2-day) plants. However, a low Pi (167 juveniles/ plant) resulted in root proliferation on 4-day-old plants at 26 C. Mi was most damaging in a low clay content soil mixture. PMID- 19294186 TI - Relative Virulence of Meloidogyne incognita Host Races on Soybean. AB - Sensitivity and host efficiency of susceptible ('Lee 68', 'Coker 156') and resistant ('Bragg', 'Centennial', 'Forrest', 'Lee 74') soybean (Glycine max (L.) Merr.) cultivars for races of Meloidogyne incognita (Mi) were determined in greenhouse experiments. Eight Mi populations collected from the southeastern United States were utilized. All Mi races reproduced readily on Lee 68 and Lee 74 and moderately on Forrest and Bragg. Coker 156 exhibited resistance to races 1 and 2, and some race 3 populations, but was very susceptible to certain race 3 and 4 populations. Reproduction of all races was lowest on Centennial. Forrest and Centennial shoot growth was not significantly suppressed by any race. There were no distinct differences in virulence between races except for a race 3 population which reproduced readily on all cultivars, stunting their growth. Considerable variation in reproduction existed within races 1 and 3. PMID- 19294187 TI - Effects of Soil Type on the Damage Potential of Meloidogyne incognita on Soybean. AB - Effects of soil type on the reproduction and damage potential of Meloidogyne incognita on soybean, Glycine max (L.) Merr., were determined at five locations in North Carolina, including one site where plots with six soil types were established. M. incognita reproduced readily on a susceptible soybean cultivar in most soil types, with somewhat limited reproduction in muck soils. The relationship between initial population densities and yield varied among soil types and nematode populations. Yield losses were greatest in sandy and muck soil types, with less nematode damage occurring in the clay soil types. A North Carolina and a Georgia population of M. incognita differed greatly in their ability to reproduce on soybean and suppress growth. The North Carolina population had a moderate effect on yield in 1981 and only a slight effect in 1982. In contrast, a Georgia population severely limited soybean growth and yield at lower initial population densities in 1983, Initial population densities of the nematodes and physical and chemical edaphic factors accounted for much of the variation of soybean yield and nematode reproduction. PMID- 19294188 TI - Effects of Environments, Meloidogyne incognita Inoculum Levels, and Glycine max Genotype on Root-knot Nematode-Soybean Interactions in Field Microplots. AB - Five soybean cultivars (Braxton, Gordon, Jeff, Bragg, and Wright) resistant to Meloidogyne incognita (Mi) and three susceptible cultivars (Coker 156, GaSoy 17, and Coker 237) were grown at two locations for four seasons in microplots with increasing initial soil population densities (Pi) of Mi. The resistant cultivars and Coker 156 yielded better than GaSoy 17 and Coker 237 at all Pi. Yield response was dependent on environmental conditions and at one location was stimulated on Braxton, Gordon, Jeff, and Bragg by low Pi. Although Mi reproduced well on all cultivars, the pattern of reproduction differed. Population densities of Mi leveled off after 90 days on GaSoy 17 and Coker 237 but were still increasing after 120 days on the resistant cultivars; population densities were lower on resistant than on the susceptible cultivars. The population density of Mi on Coker 156 after 120 days was intermediate between those on the other susceptible and on the resistant cultivars. Mi population densities followed the same pattern under varying environmental conditions. PMID- 19294189 TI - Differential Reaction of Alfalfa Cultivars to Meloidogyne hapla and M. chitwoodi Populations. AB - Meloidogyne hapla reproduced and suppressed growth (P < 0.05) of susceptible Lahontan and Moapa alfalfa at 15, 20, and 25 C. At 30 C, resistant Nevada Syn XX lost resistance to M. hapla. M. hapla invaded and reproduced on Rhizobium meliloti nodules of Lahontan and Moapa, inducing giant cell formation and structural disorder of vascular bundles of nodules without disrupting bacteroids. At 15, 20, and 25 C a M. chitwoodi population from Utah reproduced on Lahontan, Moapa, and Nevada Syn XX alfalfa, suppressing growth (P < 0.05). Final densities of the Utah M. chitwoodi population were greater (P < 0.05) than those of Idaho and Washington State populations on Lahontan at 15 and 25 C and on Nevada Syn XX at 15 C, but were less consistent and smaller (P < 0.05) than those of M. hapla on Lahontan and Moapa at 20 and 25 C. Inconsistent reproduction of the Utah M. chitwoodi population on alfalfa suggests the possible existence of nematode strains revealed by variability in alfalfa resistance. No reproduction or inconsistent final nematode population densities with no damage were observed on Lahontan, Moapa, and Nevada Syn XX plants grown in soil infested with Idaho and Washington State M. chitwoodi populations. PMID- 19294190 TI - Meloidogyne incognita Resistance Characteristics in Tomato Genotypes Developed for Processing. AB - Nine resistant processing tomato (Lycopersicon esculentum) cultivars and advanced lines were compared with four susceptible cultivars in 1,3-dichloropropene fumigated and nontreated plots on Meloidogyne incognita-infested sites over 3 years. Yield of all resistant genotypes grown in nontreated and nematicide treated plots did not differ and was greater than yield of susceptible genotypes. M. incognita initial soil population densities caused 39.3-56.5% significant (P = 0.05) yield suppressions of susceptible genotypes. Nematode injury to susceptible plants usually caused both fruit soluble solids content and pH to increase significantly (P = 0.05). Only trace nematode reproduction occurred on resistant genotypes in nontreated plots, whereas large population density increases occurred on susceptible genotypes. Slightly greater nematode reproduction occurred on resistant genotypes at the southern desert location, where soil temperature exceeded 30 C, than at other locations. At two locations resistant MOX 3076 supported greater reproduction than other resistant genotypes. PMID- 19294191 TI - Control of Heterodera carotae, Ditylenchus dipsaci, and Meloidogyne javanica with Fumigant and Nonfumigant Nematicides. AB - Five field trials were conducted in Italy in 1983 and 1984 to test the efficacy of isazofos and benfuracarb in controlling Heterodera carotae on carrot, Ditylenchus dipsaci on onion, and Meloidogyne javanica on tomato. Methyl isothiocyanate (MIT) was tested against H. carotae and M. javanica. Single (10 kg a.i./ha) and split (5 + 5 kg a.i./ha) applications of isazofos gave yield increases of carrot and onion similar to those obtained with DD (300 liters/ha) and aldicarb (10 kg a.i./ha). Population densities of H. carotae in carrot roots at harvest and of M. javanica in tomato roots 2 months after transplanting were also suppressed by isazofos. Benfuracarb (10 kg a.i./ha increased onion yields in a field infested with D. dipsaci, but it was not effective against H. carotae or M. javanica. The efficacy of MIT at 400 and 600 liters/ha was similar to that of MIT + DD (Di-Trapex) at 300 liters/ha. Both nematicides inhibited hatch of H. carotae eggs and decreased the soil population density of M. javanica. PMID- 19294192 TI - Management of Root-knot Nematodes by Phenamiphos Applied through an Irrigation Simulator with Various Amounts of Water. AB - Phenamiphos (6.7 kg a.i./ha) was applied via an irrigation simulator to squash at planting (AP) and 2 weeks after planting (PP), and to corn AP and 1 week PP to manage root-knot nematodes (Meloidogyne incognita). The nematicide was applied with 0.25, 0.64, 1.27, and 1.91 cm surface water/ ha to a Lakeland sand in which the soil moisture was at or near field capacity. Based on efficacy and crop response, no additional benefits resulted when phenamiphos was applied in volumes of water greater than 0.25 crn/ha. The cost of applying each 0.25 cm of water over a hectare is approximately $1.08, or a 92% reduction in nematicide application cost over conventional methods ($13.50/ha). Low root-gall indices and high yields from squash and corn indicate more effective nematode management when phenamiphos was applied AP rather than PP. Results from this method of applying phenamiphos suggest that certain nematicides could be used as salvage alternatives when nematodes are detected in crops soon after planting. For multiple-pest management, nematicides, other compatible biocides, and fertilizers could be applied simultaneously with sprinkler irrigation. PMID- 19294193 TI - Heterodera glycines-Soybean Association: A Rapid Assay Using Prun Seedlings. AB - A 16-day bioassay to determine compatible and incompatible associations of soybeans and cyst nematodes is described. This permits large-scale experiments in the laboratory. Seedlings are placed between two sheets of moist paper towelling so that the root tips are even with the bottom edge. The towelling is then rolled and set on the surface of cyst-infested soil. Radicles are permitted to grow through the soil for 24 hours after which cotyledons and apical portions are trimmed to retard root growth. The plants are assembled in bundles and maintained in aerated water in test tubes for 15 days before counts of emerged nematodes are taken. This system is economical in both space and labor. PMID- 19294194 TI - Effects of Continuous Cropping of Resistant and Susceptible Cultivars on Reproduction Potentials of Heterodera glycines and Globodera tabacum solanacearum. AB - The reproductive potentials of Heterodera glycines (mixture of races 3 and 4 and unidentified races) and a tobacco cyst nematode Globodera tabacum solanacearum were studied in the field. The experiments involved four cultivars of soybean Glycine max and four cultivars of Nicotiana tabacum. The reproductive potential of the H. glycines population was high on Essex and Lee 74 soybean but low on Forrest and Bedford over the 3 years (1982-84) of continuous cropping. The reproductive potential of H. glycines was 12% on Forrest and 6% on Bedford in 1982 but increased to 37 and 35% in 1983 and to 71 and 41% in 1984, respectively, on these two cultivars. The reproductive potential of G. tabacum solanacearum was high on McNair 944 and Coker 319 tobacco cultivars and low on VA 81 and PD 4 over the 3 years of cropping. The reproductive potential of G. tabacum solanacearum on VA 81 and PD 4 was 18 and 17% in 1982, 7 and 16% in 1983, and 5 and 5% in 1984, respectively. The changes in reproductive potentials of H. glycines and G. tabacum solanacearum may be related to inherent genetic variability in the systems that control reproduction of the two cyst nematodes and nature of resistance incorporated in the soybean and tobacco cultivars. PMID- 19294195 TI - Relationship between Root Growth of Potato, Root Diffusate Production, and Hatching of Globodera rostochiensis. AB - Hatching response of Globodera rostochiensis in potato root diffusate (PRD) collected by soaking individual potato, Solanum tuberosum, root systems in water for 2 hours was used to assess the relationship between root growth and PRD production. Resistant potato cultivars Hudson and Rosa were used as test plants. Maximum hatch occurred in PRD collected 3 weeks after plant emergence (AE) in the greenhouse, and declined after this time. Hatch was positively correlated with increased root weight only during the first 3 weeks AE. Hudson PRD was consistently more active than Rosa PRD in stimulating hatch, except when adjusted for root weight. Although the results indicated that cells at the root tip produced a more active PRD than cells located elsewhere, PRD appeared to be produced along the entire root. Differences in time length of the vegetative growth phase, extent of root growth, and volume of roots, rather than the production of a more active PRD per se, may explain why Hudson is more effective than Rosa in reducing G. rostochiensis population densities in soil. PMID- 19294196 TI - Influence of Pratylenchus penetrans on Plant Growth and Water Relations in Potato. AB - Plants of potato (Solanum tuberosum) cultivars Katahdin and Superior were inoculated with 0, 1,500, or 15,000 Pratylenchus penetrans. Transpiration, measured in the greenhouse with a porometer after 56 days of growth, was not significantly different among nematode inoculum levels or between cultivars. The rate of xylem exudation from decapitated root systems of Katahdin plants inoculated with 1,500 or 15,000 P. penetrans and Superior plants inoculated with 15,000 P. penetrans was lower than from noninoculated plants. Root weight of Katahdin and Superior was not affected by P. penetrans inoculum level. Transpiration of plants inoculated with 0, 500, 5,000 or 50,000 P. penetrans was recorded weekly from 14 to 56 days after planting. No consistent effects of nematode inoculum density on transpiration rate were observed. Root hydraulic conductivity was lower in Katahdin plants inoculated with 266 P. penetrans per plant and in Chippewa with 5,081 per plant than in noninoculated plants. Nematodes reduced leaf area of Superior, Chippewa, and Katahdin and root dry weight of Chippewa but had no effect on growth of Hudson, Onaway, or Russet Burbank plants. Assessing nematode effects on root hydraulic conductivity may provide a measure of the tolerance of potato cultivars to nematodes. PMID- 19294197 TI - Population Dynamics, Root Penetration, and Feeding Behavior of Pratylenchus agilis in Monoxenic Root Cultures of Corn, Tomato, and Soybean. AB - Population dynamics, rate of root penetration, and external root feeding behavior of Pratylenchus agilis (Pa) in monoxenic cultures of intact corn seedlings and root explants of corn, tomato, and soybean were studied. In descending order of suitability as hosts were I. O. Chief corn, Rutgers tomato, and Williams soybean. Soybean entries Kent, Pickett 71, PI 90763, and Essex were poor hosts. Numbers of eggs and vermiform Pa in the agar medium indicated total fecundity and host suitability. Agar, sand, or soil as support media did not appear to affect Pa root penetration, but the rate of corn root growth did. Whereas most vermiform Pa and eggs were in roots, substantial numbers appeared able to feed and complete their life cycle as ectoparasites on root epidermal cells and root hairs. PMID- 19294198 TI - Cambium destruction in conifers caused by pinewood nematodes. AB - Percentage and rate of mortality in 2-4-year-old conifers depended upon the numbers of pinewood nematodes Bursaphelenchus xylophilus inoculated into their stems. In addition, percentage of conifer mortality was greater for spring inoculations when cambial activity was greater than for late summer and fall inoculations. Gross and histological examination of stems revealed destruction of the cambial layer, including fusiform and ray intitials and their derivatives. These data suggest that cambial and ray destruction causes tree death through blockage of tracheids by gas, oleoresin, or metabolites from dying ray tissues. PMID- 19294199 TI - Nematode trophic structure in conventional and no-tillage agroecosystems. AB - The effect of tillage intensity on nematode community trophic structure and the role of nematodes in the regulation of decomposition rates in agroecosystems were examined. Conventional (CT) and no-tillage (NT) agroecosystems were sampled monthly for 1 year. Tillage affected nematode trophic structure and total abundance. Monthly mean densities of bacterivorous, fungivorous, and total nematodes were greater in CT than in NT plots. In the summer, however, fungivorous and plant parasitic nematodes were more abundant in NT. No difference was detected for omnivore-predator nematodes. PMID- 19294200 TI - Penetration, Development, Reproduction, and Sex Ratio of Meloidogyne javanica in Three Carrot Cultivars. PMID- 19294201 TI - Comparison of Development, Reproduction, and Aggressiveness of Meloidogyne incognita Races 3 and 4 on Cotton. PMID- 19294203 TI - A new, efficient technique for permanent nematode storage. PMID- 19294202 TI - Methods for Reducing Experimental Variation in Globodera rostochiensis. PMID- 19294204 TI - A simple and reliable device for monitoring fumigant residues in soil. PMID- 19294205 TI - A simple, inexpensive, portable apparatus for injecting experimental chemicals in drip irrigation systems. PMID- 19294206 TI - Transfer of type specimens. PMID- 19294207 TI - Penetration and Postinfection Development of Meloidogyne incognita on Cotton as Affected by Glomus intraradices and Phosphorus. AB - The influence of the vesicular-arbuscular mycorrhizal fungus Glomus intraradices (Gi) and superphosphate (P) on penetration, development, and reproduction of Meloidogyne incognita (Mi) was studied on the Mi-susceptible cotton cultivar Stoneville 213 in an environmental chamber at 28 C. Plants were inoculated with Mi eggs at planting or after 28 days and destructively sampled 7, 14, 21, and 28 days after nematode inoculation. Mi penetration after 7 days was similar in all treatments at either inoculation interval. At 28 days, however, nematode numbers were least in mycorrhizal root systems and greatest in root systems grown with supplemental P. The rate of development of second-stage juveniles to ovipositing females was unaffected by Gi or P when Mi was added at planting, but was delayed in mycorrhizal root systems when Mi was added 28 days after planting. Nematode reproduction was lower in mycorrhizal than in nonmycorrhizal root systems at both Mi inoculation intervals. Nematode reproduction was stimulated by P when Mi was added at planting, but was similar to reproduction in the low P nonmycorrhizal treatment when Mi was added 28 days after planting. Eggs per female were increased by P fertility when Mi was added at planting. PMID- 19294208 TI - Effects of Interactions among Heterodera glycines, Meloidogyne incognita, and Host Genotype on Soybean Yield and Nematode Population Densities. AB - The effects of host genotype and initial nematode population densities (Pi) on yield of soybean and soil population densities of Heterodera glycines (Hg) race 3 and Meloidogyne incognita (Mi) race 3 were studied in a greenhouse and field microplots in 1983 and 1984. Centennial (resistant to Hg and Mi), Braxton (resistant to Mi, susceptible to Hg), and Coker 237 (susceptible to Hg and Mi) were planted in soil infested with 0, 31, or 124 eggs of Hg and Mi, individually and in all combinations, per 100 cm(3) soil. Yield responses of the soybean cultivars to individual and combined infestations of Hg and Mi were primarily dependent on soybean resistance or susceptibility to each species separately. Yield of Centennial was stimulated or unaffected by nematode treatments, yield of Braxton was suppressed by Hg only, and yield suppressions caused by Hg and Mi were additive and dependent on Pi for Coker 237. Other plant responses to nematodes were also dependent on host resistance or susceptibility. Population densities of Mi second-stage juveniles (J2) in soil were related to Mi Pi and remained constant in the presence of Hg for all three cultivars. Population densities of Hg J2 on the two Hg-susceptible Cultivars, Braxton and Coker 237, were suppressed in the presence of Mi at low Hg Pi. PMID- 19294209 TI - Effects of Heterodera glycines and Meloidogyne incognita on Early Growth of Soybean. AB - Greenhouse and field microplot studies were conducted to compare soybean shoot and root growth responses to root penetration by Heterodera glycines (Hg) and Meloidogyne incognita (Mi) individually and in combination. Soybean cultivars Centennial (resistant to Hg and Mi), Braxton (resistant to Mi, susceptible to Hg), and Coker 237 (susceptible to Hg and Mi) were selected for study. In the greenhouse, pot size and number of plants per pot had no effect on Hg or Mi penetration of Coker 237 roots; root weight was higher in the presence of either nematode species compared with the noninoculated controls. In greenhouse studies using a sand or soil medium, and in field microplot studies, each cultivar was grown with increasing initial population densities (Pi) of Hg or Mi. Interactions between Hg and Mi did not affect early plant growth or number of nematodes penetrating roots. Root penetration was the only response related to Pi. Mi penetration was higher in sand than in soil, and higher in the greenhouse than in the field, whereas Hg penetration was similar under all conditions. At 14 days after planting, more second-stage juveniles were present in roots of susceptible than in roots of resistant plants. Roots continued to lengthen in the greenhouse in the presence of either Mi or Hg regardless of host genotype, but only in the presence of Mi in microplots; otherwise, responses in field and greenhouse studies were similar and differed only in magnitude and variability. PMID- 19294210 TI - Soybean and Maize Cropping Models for the Management of Meloidogyne incognita in the Coastal Plain. AB - Models are presented to describe the influence of rotations of Meloidogyne incognita-susceptible cultivars, resistant cultivars, and maize on postharvest abundance of M. incognita juveniles in the soil. Depending on initial densities of juveniles, monocultured regimes reached equilibrium densities after a few years of 287, 40, and 10 juveniles per 10 cm(3) soil for susceptible soybean, resistant soybean, and maize, respectively. Yearly changes in the population density of juveniles due to rotation of these crops were simulated by iterative substitution of the model equations for each crop. A maximum density of 319 per 10 cm(3) soil was reached following a susceptible cultivar in a susceptible resistant soybean rotation. Soybean yield loss estimates are presented for monocultured regimes and for various rotations with maize. PMID- 19294211 TI - Population Changes of Heterodera glycines and Soybean Yields Resulting from Soil Treatment with Alachlor, Fenamiphos, and Ethoprop. AB - The population dynamics of Heterodera glycines as influenced by alachlor, fenamiphos, and ethoprop alone and in herbicide-nematicide combinations were studied in the field. Numbers of H. glycines juveniles and eggs were higher at midseason and harvest where nematicides were applied. Fenamiphos alone or in combination with alachlor provided better control of H. glycines and greater seed yields than treatments with ethoprop. Numbers of H. glycines eggs at harvest in 1980 were positively correlated with numbers of juveniles at planting in 1981 and negatively related to seed yield in 1981. PMID- 19294212 TI - Soybean yield as related to rates of 1,3-dichloropropene applied at planting for management of root-knot disease. AB - 1,3-Dichloropropene (1,3-D) at rates of 17.2 to 51.6 liters/ha applied 3 days preplant or at planting significantly (P < 0.05) reduced the amount of galling on roots of soybean grown in sites infested with Meloidogyne incognita or M. arenaria. Populations of M. incognita second-stage juveniles at harvest were significantly (P < 0.05) reduced by all treatments. Only the 51.6-liters/ ha treatments and a 3-day preplant 34.4-liters/ha application significantly reduced at-harvest juvenile infestations of M. arenaria. Equations (P < 0.001) relating soybean yield and 1,3-D dosage indicated soybean phytotoxicity at the upper range of the nematicide rates. The maximum yield response was predicted at 40 liters/ha applied 3 days preplant at both infestation sites. Maximum yield response was predicted with 30 liters/ha applied at planting to M. incognita-infested soil and from 25 liters/ha applied at planting to M. arenaria-infested soil. Application of economic factors suggested that management of M. incognita may be cost effective with at-plant treatments of low rates of 1,3-D. Yield responses of M. arenaria-infected soybean exposed to similar treatments were insufficient to justify their use at prevailing prices. PMID- 19294213 TI - Effects of Temperature on Development of Heterodera glycines on Glycine max and Phaseolus vulgaris. AB - Soybean cyst nematode resistant 'Fayette' and susceptible 'Williams 79' soybeans (Glycine max) and resistant 'WIS (RRR) 36' and susceptible 'Eagle' snap beans (Phaseolus vulgaris) were used in determining the effects of host and temperature on the development, female production, sex ratios, and host response to Heterodera glycines. Temperatures were maintained constant at 16, 20, 24, 28, and 32 C using water-filled tanks. The most rapid development and greatest female production occurred between 20 and 28 C. The equation DS = 5(10)x(2)y(2) - 3(10)x(2)y - 2.8(10(3))x(2) - 1.94(10(2))y(2) + 0.4288x + 1.0220y - 12.7185, where DS = developmental stage, X = time, and Y = temperature, predicted the developmental stage of the nematode and accounted for 84% of the variation. Male : female ratios did not differ within this range and were generally less than one. At all temperatures the resistant soybean produced the greatest number of necrotic responses to H. glycines infection, followed by the resistant snap bean. The susceptible soybean and snap bean produced the fewest necrotic responses. PMID- 19294214 TI - Scanning Electron Microscopy of Second-Stage Juvenile Cephalic Morphology in Heterodera glycines Races. AB - Scanning electron microscopy (SEM) was used to compare juvenile cephalic morphology of the five described races of Heterodera glycines. Races 1, 2, 3, and 4 were obtained in the United States and race 5 was obtained from Japan. Differences in the gross morphology o f labial discs; ventral, dorsal and lateral lips; amphidial apertures; and fissures on the labial disc o f some specimens were observed. There was considerable interracial and intraracial variation which precluded separation o f juveniles of H. glycines races by SEM. PMID- 19294215 TI - A comparison of techniques useful for preparing nematodes for scanning electron microscopy. AB - Second-stage juveniles of Meloidogyne incognita were prepared by several different techniques for scanning electron microscopy (SEM). Sequential fixation in the cold (4-8 C) was superior to rapid fixation at room temperature, glutaraldehyde and glutaraldehyde-formalin were better fixatives than formalin alone, and critical point drying with carbon dioxide or Freon gave similar results that were only slightly better than air drying with Freon. Freeze drying sequentially fixed nematodes from 100% ethanol in liquid propane produced the best preserved specimens with the fewest artifacts. Specimens of various free living and plant-parasitic nematodes were prepared for SEM by freeze drying. This technique was adequate for most genera but unsatisfactory for a few. Although each genus may require a different procedure for optimum preservation of detail, sequential fixation with glutaraldehyde and freeze drying are comparable and often superior to commonly used techniques for preparing nematodes for SEM. PMID- 19294216 TI - Effect of Meloidogyne incognita and Importance of the Inoculum on the Yield of Eggplant. AB - The relationship between population densities of race 1 of Meloidogyne incognita and yield of eggplant was studied. Microplots were infested with finely chopped nematode-infected pepper roots to give population densities of 0, 0.062, 0.125, 0.25, 0.50, 1, 2, 4, 8, 16, 32, 64, and 128 eggs and juveniles/cm(3) soil. Both plant growth and yield were suppressed by the nematode. A tolerance limit of 0.054 eggs and juveniles/cm(3) soil and a minimum relative yield of 0.05 at four or more eggs and juveniles/cm(3) soil were derived by fitting the data with the equation y = m + (1 - m)z(PT). Maximum nematode reproduction rate was 12,300. Hatch of eggs from egg masses in water or from sodium hypochlorite dissolved egg masses was similar (41% and 39%), but egg viability was significantly greater from egg masses in water (58%) than from sodium hypochlorite dissolved egg masses (12%) after 4 weeks. Greater numbers of nematodes were collected from roots of tomatoes from soil infested with entire egg masses than from tomato roots from soil infested with egg masses dissolved by sodium hypochlorite. PMID- 19294217 TI - Retention of Resistance to Meloidogyne incognita in Lycopersicon Genotypes at High Soil Temperature. AB - Lycopersicon glandulosum and L. peruvianum clones and L. esculentum cultivars 'VFN8' (resistant) and 'Rutgers' (susceptible) were tested for their resistance to Meloidogyne incognita (race l) at soil temperatures of 25 and 32 C. L. esculentum cv. VFN8 and L. peruvianum Acc. No. 128657, both of which possess the Mi gene, were resistant at 25 C but were susceptible at 32 C. L. glandulosum Acc. No. 126443 and L. peruvianum Acc. No. 270435, with combined resistance to M. hapla and M. incognita, and L. peruvianum Acc. Nos. 129152 and LA2157, with resistance to M. incognita, were highly resistant at both temperatures. In a second experiment three of these accessions under heat stress simulated by 32 C ambient and soil temperature retained a high level of resistance. Two clones of L. glandulosum Acc. No. 126440, with resistance to M. hapla, were moderately susceptible to M. incognita at 25 and highly susceptible at 32 C. M. incognita produced significantly (P = 0.01) more eggs on L. esculentum cv. Rutgers at 32 than at 25 C. This study supports the existence of genes other than the Mi gene that confer resistance to M. incognita and are functional at high soil temperatures. PMID- 19294218 TI - Varietal Response and Estimates of Heritability of Resistance to Meloidogyne javanica in Carrots. AB - With methods developed in this study, varietal responses to M. javanica were evaluated and heritability of resistance of two promising carrot cultivars was estimated. More egg masses were found on root systems inoculated with eggs added to the soil in three holes in 250 cm(3) cups than by mixing the inoculum with soil in the cups. A resistant breeding line, CNPH 1437, was discriminated from susceptible cultivar Nova Kuroda with inoculum levels higher than 2,000 eggs per cup. Greenhouse and field results suggested that cultivars Nantes Superior and Shin Kuroda were susceptible, Kuronan was somewhat tolerant, and Brasilia and Tropical were resistant to M. javanica. Nantes Superior or Shin Kuroda yielded less in carbofuran-treated soil (3 kg a.i./ha) than Kuronan, Brasilia, and Tropical did in nontreated soil. However, incorporation of the nematicide greatly increased yields of Kuronan (32%), Brasilia (62%), and Tropical (91%). Primary root galling at the seedling stage was an adequate parameter for resistance evaluation. Estimated heritability were 0.48 +/- 0.07 for primary root galling and 0.35 +/- 0.08 for egg mass production in Brasilia, and 0.16 +/- 0.11 for primary root galling and 0.31 +/- 0.09 for egg mass production in Kuronan. PMID- 19294219 TI - Root Growth of Susceptible and Resistant Potato Cultivars and Population Dynamics of Globodera rostochiensis in the Field. AB - Globodera rostochiensis population densities and potato root growth were measured in field plots of one susceptible and two resistant potato cultivars. Root growth and nematode densities were estimated from soil samples taken at three depths between plants within the rows, three depths 22.5 cm from the rows, and at two depths midway between rows (furrows). Four weeks after plant emergence (AE), nematode densities in the rows had declined 68% in plots of the susceptible cultivar and up to 75% in plots of both resistant cultivars. Significant decline in nematode densities in the furrows 4 weeks AE occurred only in plots of the susceptible cultivar. Total decline in nematode density in fallow soil was 50%, whereas in plots of the resistant cultivars, decline was more than 70% in the rows and more than 50% in the furrows. Nematode densities increased in the rows of the susceptible cultivar but declined in the furrows. We conclude that G. rostochiensis decline or increase is correlated with host resistance and the amount of roots present at any particular site. PMID- 19294220 TI - Influence of Alfalfa Plant Growth on the Multiplication Rates and Ceiling Population Density of Meloidogyne hapla. AB - The rates of reproduction and multiplication of Meloidogyne hapla decreased as a result of self-regulatory, density-dependent processes with time and nematode population increase in the soil and roots of Medicago sativa cv. Cuf 101. Juvenile, egg, and mature female population densities increased at a maximum rate until damage to the host resulted in alfalfa yield reductions. Temporal differences in multiplication and reproduction rates of M. hapla were observed to be a function of initial population density (Pi), host damage, and root biomass, indicating increased levels of competition for a constant but limited number of feeding sites. Over time, a log linear relationship emerged between multiplication rate of M. hapla and Pi. Slopes of -0.90953 for combined eggs and juveniles and -0.71349 for mature females indicated a gradual approach to ceiling densities. Reproductive rates decreased exponentially from an initial maximal value of 200 to a relatively constant rate of 53 eggs per female. PMID- 19294221 TI - A Simulation Model of Heterodera schachtii Infecting Beta vulgaris. AB - A simulation model of a single sugarbeet, Beta vulgaris L., plant infected by the sugarbeet cyst nematode, Heterodera schachtii Schmidt, was developed using published information. The model is an interactive computer simulation programmed in FORTRAN. Given initial population densities of the nematode at planting, the model simulates nematode population dynamics and the growth of plant tap and fibrous roots. The driving variable for nematode development and plant growth is temperature. PMID- 19294222 TI - Meloidogyne hispanica n. sp. (Nematoda: Meloidogynidae), the 'Seville Root-Knot Nematode'. AB - Meloidogyne hispanica n. sp. is described and illustrated from specimens obtained from peach rootstock, Prunus persica silvestris Batsch, from the Seville district of Spain. The perineal pattern of the female is oval shaped to rectangular with low dorsal arch and often widely spaced lateral lines with fringe-like striae. The stylet, 14.1 mum long, has broad, distinctly set off knobs. Males have a high, rounded head cap that slopes posteriorly. Labial disc and medial lips are fused to form elongate lip structures. The robust styler, 23.5 mum long, has large, rounded knobs that are slightly set off from the shaft. Mean second-stage juveniles length is 392.6 mum. The truncate head region is generally not annulated. The distinctly rounded and raised labial disc and the crescent-shaped medial lips form dumbbell-shaped lip structures. The stylet, 11.1 mum long, has rounded, posteriorly sloping knobs. The slender tail, 46.4 mum long, has large irregular-sized annules in the posterior region and ends in a bluntly rounded tip. Tomato was a good host; tobacco, pepper, and watermelon were poor hosts; cotton and peanut were nonhosts. Meloidogyne hispanica n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of 2n = 33-36. The esterase pattern is unique among Meloidogyne species. PMID- 19294223 TI - Description of Meloidogyne christiei n. sp. (Nematoda: Meloidogynidae) from Oak with SEM and Host-Range Observations. AB - Meloidogyne christiei n. sp. is described and illustrated from turkey oak (Quercus laevis) in Sanlando Park, Altamonte Springs, Florida. This new nematode species has a distinctive perineal pattern commonly with a high, squarish arch and coarse broken striae which tend to diverge at various angles, especially in and above the anal area. Female labial disc is indented, forming four points or prongs, unlike other species. Eggs are deposited inside the gall in a tubular, coiled manner. Vaginal muscles are exceptionally prominent and dense. SEM observations provided further detail of the perineal pattern and details of the head of females, males, and second-stage juveniles. Galls on the root commonly occur singly, but sometimes in small clusters, and appear as discrete nodules on the side of the root and without adjacent swelling. In general, only one female is found in each gall but occasionally two are present. In greenhouse tests, citrus, tobacco, cotton, pepper, watermelon, peanut, and tomato were not hosts. This nematode occurs throughout central Florida commonly on Q. laevis, the only known host. PMID- 19294224 TI - New Species of Cyst Nematode Heterodera pakistanensis (Nematoda: Heteroderidae) Attacking Wheat in Pakistan. AB - Heterodera pakistanensis n. sp., described and illustrated from roots of common wheat (Triticum aestivum) from Sukkur, Sind, Pakistan, belongs to the goettingiana group. It is most closely related to H. cyperi Golden, Rau &Cobb, 1962, H. raskii Basnet & Jayaprakash, 1984, and H. mothi Khan &Husain, 1965. Second-stage juveniles (J2) can be distinguished from H. cyperi J2 by an areolated lateral field with four incisures and shorter stylet, whereas cysts are separated by a more elongated vulva slit and the conspicuous structure of the underbridge. It differs from H. raskii by having four areolated lateral lines in J2, smaller female lemon-shaped cyst, shorter fenestra length and width, conspicuous underbridge, and distinct anus with a high cuticular pattern 40-45 mum from posterior end. It also differs from H. mothi by the presence of four areolated lateral lines in J2 and absence of vulva denticles and bullae. PMID- 19294225 TI - Description of Thecavermiculatus carolynae n. sp. (Nematoda: Ataloderinae). AB - Thecavermiculatus carolynae n. sp. from the Pacific coastal area of California is described from female, male, and juvenile (J2) specimens reared on Festuca megalura Nutt. in the greenhouse. Differences between females of T. carolynae and those of its closest relative, T. gracililancea Robbins, 1978, include shorter stylet, shorter length, and lesser length of the vulval slit. Of these two species, males have been found only for T. carolynae. The habitats of the two species are vastly different: T. gracililancea has been found only in shallow clay soil near hilltops in an area subject to high summer temperatures where the soil becomes brick-like during the summer; T. carolynae has been found only in deep sandy soil of a creek bottom subject to cool summer temperatures where the soil remains moist. PMID- 19294226 TI - Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Morphometrics and Lectotype Designation of D. heterocephalus Cobb, 1914. AB - Dolichodorus cobbi n. sp. is described and illustrated from soil around roots of "woods" on the Stanley Farm, Scott County, Arkansas. This species is distinguishable from others of the genus by its short tail projection. It is most closely related to D. marylandicus but differs in having a short tail projection, longer stylet, greater body length, larger c value, and also in position of excretory pore. A limited number of Cobb's original specimens and drawings of D. heterocephalus were examined. A lectotype and paralectotype were designated to establish the taxonomic base for the genus. Cobb's original modified labeled drawing of D. heterocephalus is also included. Morphometric data on D. heterocephalus from the type locality and several other populations are given. Some variations in female tail shape, body length, and stylet length were noted. PMID- 19294227 TI - Effects of Cyanide Ion and Hypoxia on the Volumes of Second-Stage Juveniles of Meloidogyne incognita in Polyethylene Glycol Solutions. AB - Changes in the volumes of second-stage juveniles of Meloidogyne incognita were monitored in aqueous solutions of polyethylene glycol supplemented with dilute balanced salts. At key points within a 48-hour cycle of fluctuating water potential, nematodes were placed under hypoxic conditions or exposed to the respiratory inhibitor, sodium cyanide, to detect any respiration-dependent process that regulates volume. Aerobic respiratory arrest at -500 kPa induced pronounced water loss, lateral and dorsoventral collapse of the body wall, and abnormal failure to shorten longitudinally. Durations of hypoxia that were innocuous in dilute solutions were lethal during 500 kPa increases and decreases in water potential; the same water potential changes under aerobic conditions had no effect on viability. Data are consistent with the hypothesis that respiration is essential to survive water potential changes. PMID- 19294228 TI - Fluorescent and Ferritin Labelling of Cuticle Surface Carbohydrates of Caenorhabditis elegans and Panagrellus redivivus. AB - Caenorhabditis elegans and Panagrellus redivivus were investigated for surface carbohydrates using fluorescent-labelled and ferritin-labelled lectins. Rhodamine labelled Concanavalin A was specifically located in the cephalic region of both species. Rhodamine-labelled wheat germ agglutinin was located over the entire cuticle of P. redivivus but was absent on C. elegans. Rhodamine-labelled peanut agglutinin and Limax flavus agglutinin did not label nematodes of either species. Galactose and sialic acid were not detected on either species, whereas mannose glucose residues were specifically localized in the head areas of both species. No detectable N-acetylglucosamine occurred on C. elegans, but it was evenly distributed over the cuticle surface of P. redivivus. PMID- 19294229 TI - Pinewood Nematode, Bursaphelenchus xylophilus, Associated with Red Pine, Pinus resinosa, in Western Maryland. AB - Red pines Pinus resinosa in Garrett and Allegany counties, Maryland, were examined during 1982-84 to determine distribution of the pinewood nematode, Bursaphelenchus xylophilus, within and among trees. Approximately 25-year-old (younger) and 47-year-old (older) trees were subdivided into the following categories: 1) trees with mostly green needles; 2) trees with mostly reddish brown needles; 3) trees lacking needles but with bark intact; 4) trees lacking both needles and bark; and 5) trees with chlorotic, bleached-green needles. Bursaphelenchus xylophilus was found infecting 68% of younger red pines and 77% of older red pines. Nematodes were not evenly distributed in trees within any given tree decadence category or in trees of the same age. Nematodes were recovered from 20% of wood samples from trunks and primary and secondary branches in younger pines and from 15 % of older red pines. On the basis of tree decadence category, the highest incidence of infection in younger trees (31%) was in bleached-green needled trees (category 5), whereas in older trees the highest infection (25%) occurred in green needled trees (category 1). At both sites trunks were infected more often than branches. PMID- 19294230 TI - Reproduction of Pratylenchus brachyurus on Soybean Callus Tissue: Effects of Culture Age and Observations on Anhydrobiosis. PMID- 19294231 TI - Inflorescence and Leaf Galls on Palisota barteri Caused by Meloidogyne javanica. PMID- 19294232 TI - Distribution of Ditylenchus dipsaci in Daffodil Bulbs. PMID- 19294233 TI - Detection of selective cationic amphipatic antibacterial peptides by Hidden Markov models. AB - Antibacterial peptides are researched mainly for the potential benefit they have in a variety of socially relevant diseases, used by the host to protect itself from different types of pathogenic bacteria. We used the mathematical computational method known as Hidden Markov models (HMMs) in targeting a subset of antibacterial peptides named Selective Cationic Amphipatic Antibacterial Peptides (SCAAPs). The main difference in the implementation of HMMs was focused on the detection of SCAAP using principally five physical-chemical properties for each candidate SCAAPs, instead of using the statistical information about the amino acids which form a peptide. By this method a cluster of antibacterial peptides was detected and as a result the following were found: 9 SCAAPs, 6 synthetic antibacterial peptides that belong to a subregion of Cecropin A and Magainin 2, and 19 peptides from the Cecropin A family. A scoring function was developed using HMMs as its core, uniquely employing information accessible from the databases. PMID- 19294234 TI - Localization of the attachment site of oligoglucans to Mesorhizobium loti HAMBI 1148 murein. AB - The location and nature of the linkage between peptidoglycan and oligoglucans in the cell wall of Mesorhizobium loti HAMBI 1148 have been defined by the analysis of nitrous acid deamination of peptidoglycan glucosaminyl residues. The MurNH(2) Glc(n) fraction was obtained after converting deaminoacylated and N-deacetylated muramyl residues in the cell wall preparation to lactam forms which were stable during subsequent deamination, followed by reduction and opening of the lactams. GC/MS analysis of this material, subjected to partial hydrolysis and reduction or to methanolysis followed by peracetylation, confirmed the presence of glucosyl residues glycosidically attached to muramic acid. The MALDI-TOF spectroscopic analysis of the deaminated material also revealed the presence of [M-H](-) or [M+Na-2H](-) ions representing fragments containing muramic acid with one to three linked glucose residues. The analysis of fully methylated neutral oligosaccharides released from the peptidoglycan with lysozyme followed by borohydride reduction showed the presence of di- and trisaccharides lacking the reducing end. PMID- 19294236 TI - Role of computed tomography in the evaluation of orthodontic treatment in adult patients with obstructive sleep apnea syndrome (OSA). AB - OBJECTIVE: The aim of this study was to investigate pharyngeal size differences between pre- and posttrials of a Mandibular Advancement Device (MAD), using a computed tomography (CT),in the treatment of Obstructive Sleep Apnea (OSA) adult patients. MATERIALS AND METHODS: Eighteen patients with mild to moderate OSA (mean Apnea/Hypopnea Index, AHI, of 16.7) were treated with a MAD to wear at night only. After 3 months of treatment, three-dimensional changes in pharyngeal dimensions were measured on CT images performed with a sixteen detector-row CT scanner (Light Speed Plus; GE Medical Systems). Two consecutive axial sections from the hard palate to the epiglottis were obtained with and without the appliance. Measurements were made of the following airway areas (mm2) and lengths (mm): RF (nasopharynx); ROF (naso-oropharynx); OF (oropharynx); IPF (hypopharynx); SPL (soft palate length); SPT (soft palate thickness); Rgn (retrognation)-hyoid bone; hyoid bone-C2; Rgn-C2; PhL (oropharynx length); pharynx posterior wall thickness at three level. The angle between the hard and the soft palate (APDM) was also calculated. RESULTS: AHI improved significantly (from 16.7 to 11.2) when the appliance was used. Measurements from CT scans showed statistically significant expansion in the naso-oropharynx area (RF p<.014; ROF p<.050), in the Rgn-C2 length (p<.005) and in the angle between the hard and the soft palate (APDM p<.001). CONCLUSIONS: Our findings confirm the effectiveness of MAD in the treatment of patients with mild to moderate OSA. The use of MAD significantly expands the areas of the upper airway lumen most involved in the collapse. PMID- 19294237 TI - Variability of the dental surface: effects on orthodontic treatment. AB - OBJECTIVES: The Author studied the variability of the vestibular surfaces of tooth usually involved in orthodontic treatment. MATERIALS AND METHODS: 200 model casts of Italian subject with complete permanent dentition have been analyzed studying the clinical crown dimension and the angulations of tangents drawn at 3 and 4 mm. RESULTS: The authors found differences in the vestibular surfaces curvature in the last teeth of each series, more accemptuated in the lower than in the upper arch. CONCLUSION: The conclusions of this study are related to the use of orthodontic straight wire appliances. The use of standard values with a complete torque, independently from the individuals, is based on the hypothesis of equal positioning on the tooth surface and related to the teeth axis. A variation in the torque of the upper central incisor for different kinds of straight wire appliance related to extreme variation of vestibular curvature is hypothesised. PMID- 19294238 TI - Evaluation of 3D technologies in dentistry. AB - Quality of service, in terms of improvement in patient satisfaction, is an increasingly important objective in all medical fields, and is especially imperative in orthodontics due to the high numbers of patients treated. Information technology can provide a meaningful contribution to bettering treatment processes, and we maintain that systems such as CAD, CAM and CAE, although initially conceived for industrial purposes, should be evaluated, studied and customized with a view to use in medicine. The present study aims to evaluate Reverse Engineering (RE) and Rapid Prototyping (RP) in order to define an ideal chain of advanced technological solutions to support the critical processes of orthodontic activity. PMID- 19294239 TI - Interleukin-1 gene cluster polymorphisms and gingival recessions after orthodontic treatment. AB - INTRODUCTION: Genetic polymorphisms in the interleukin-1 gene cluster have been associated with the severity of periodontal diseases featured by a variable degree of destruction of connective tissue and bone, such as periodontitis and periimplantitis. This study was aimed to investigate if a link exists between such interleukin-1 gene polymorphisms and the development of gingival recessions during orthodontic treatment in Italian children. METHODS: We evaluated, in 74 young Italian patients of both sexes, the -889 C/T polymorphism of the interleukin-1alpha gene and the -511 C/T and +3954 C/T polymorphisms of interleukin-1alpha gene by polymerase chain reactions-restriction fragment length polymorphism method using NcoI, AvaI and TaqI as restriction enzymes. RESULTS: No association of interleukin-1 genotypes investigated and gingival recession occurring during orthodontic treatment were identified. CONCLUSION: In the population studied specific interleukin-1 genotypes (linked to a higher susceptibility to bone resorption in periodontal disease) there does not appear to be any association with the development of gingival recessions during orthodontic treatment. PMID- 19294240 TI - Disc-temporal adhesion and conservative functional approach: clinical evidence. AB - The Authors show the formative mechanism of disco-temporal adhesion which is a frequent disorder difficult to detect. The mechanisms of articular tension and compression are possible, possible etiopathogenetic factors. In order to point out as the integration of diagnostic and therapeutic methods can frequently provide satisfactory results in the conservative ambit, two clinical cases are described. Diagnostic-therapy iter must be individually planned according to the anatomo-functional situation and to the requests of the patient. Clinical monitoring, diagnostic and therapeutic attention and refinement are the rudiments in the approach to these problems. PMID- 19294241 TI - An unusual complication encountered during orthodontic treatment using a twin block appliance. AB - A case report is presented of a child who developed an unusual complication- lower centerline discrepancy--during Twin Block Appliance treatment due to posturing on the acrylic blocks. PMID- 19294242 TI - The Bracketless Fixed Orthodontics: nine years of clinical experimentation. AB - The Bracketless Fixed Orthodontics (BFO) is an innovative appliance made up of wires and resin. It comes from the observation of the instable position of frontal teeth still bonded to a fixed retainer and from a period of nine year of clinical experimentation on a way to preactivate a wire to obtain a wished dental movements without brackets. For this historical reasons the first experimental name was: Preactivated Retainers. It can be used to levelling teeth, as in case of relapse, or to remove the brackets before the end of the therapy. But it can be used also in more complicated clinical cases in patient that never have been treated with orthodontic therapy, alone or associated to other appliances. In this article are showed only some exemplificative clinical cases. The resolution of crowded teeth, the torque correction and the closing diastema. This appliance applied whether on the buccal aspect or on the lingual aspect of the teeth offers different advantages, as it guarantees a good control of the dental movement, also in the root movement, does not interfere in patients' phonetics and does not interfere in oral hygiene. Since don't exist neither precise references, like brackets, nor a codified therapeutic sequence the best results could be achieved accurately applying the principles of biomechanics described by Burstone and Melsen in their segmented approach. PMID- 19294243 TI - Mass spectrometric screening of chiral catalysts and catalyst mixtures. AB - The use of quasi-enantiomeric substrates and ESI-MS as an analytical tool has made it possible to determine the intrinsic enantioselectivity of chiral catalysts by monitoring catalytic intermediates. In this way, potential problems of methods based on product analysis, which may be caused by catalytically active impurities, partial dissociation of a chiral ligand-metal complex, or a non catalytic background reaction can be avoided. ESI-MS-based screening is fast, reliable, and operationally simple, as it does not require work-up or purification steps. Moreover, mixtures of catalysts with different molecular masses can be screened simultaneously, which is not possible with methods relying on product analysis. In this way catalyst libraries prepared in one batch by combinatorial methods can be screened without the need to synthesize and purify the catalysts individually. This screening method was successfully applied to Pd catalyzed allylic substitutions and metal-catalyzed and organocatalytic Diels Alder reactions. PMID- 19294244 TI - The role of electrophilic species in the Fischer-Tropsch reaction. AB - The heterogeneously catalysed Fischer-Tropsch (FT) synthesis converts syngas (CO+H2) into long chain hydrocarbons and is a key step in the economically important transformation of natural gas, coal, or biomass into liquid fuels, such as diesel. Catalyst surface studies indicate that the FT reaction starts when CO is activated at imperfections on the surfaces of late transition metals (Fe, Ru, Co, or Rh) and at interfaces with "islands" of promoters (Lewis acid oxides such as alumina or titania). Activation involves CO cleavage to generate a surface carbide, C(ad), which is sequentially hydrogenated to CHx(ad) species (x=1-4). An overview of practical aspects of the FT synthesis is followed by a discussion of the chief mechanisms that have been proposed for the formation of 1-alkenes by polymerisation of surface C1 species. These mechanisms have traditionally postulated rather non-polar intermediates, such as CH2(ad) and CH3(ad). However, electrophiles and nucleophiles are well-known to play key roles in the reactions of organic and organometallic compounds, and also in many reactions homogeneously catalysed by soluble metal complexes, including olefin polymerisation. We have now extended these concepts to the Fischer-Tropsch reaction, and show that the polymerisation reactions at polarising surfaces, such as oxide-metal interfaces, can be understood if the reactive chain carrier is an electrophilic species, such as the cationic methylidyne, CH(delta+)(ad). It is proposed that the key coupling step in C-C bond formation involves the interaction of the electrophilic methylidyne with an alkylidene (RCH(ad), R=H, alkyl), followed by an H-transfer to generate the homologous alkylidene: CHdelta+(ad)+RCH(ad)-->RCHCH(ad) and RCHCH(ad)+H(ad)-->RCH2CH(ad). If the reactions occur on non-polarising surfaces, an alternative C-C bond forming reaction such as the alkenyl+methylene, RCH=CH(ad)+CH2(ad)-->RCH=CHCH2(ad), can take place. This approach explains important aspects of the enigmatic Fischer-Tropsch reaction, and allows new predictions. PMID- 19294245 TI - Electrochemical detection of hepatitis C virus based on site-specific DNA cleavage of BamHI endonuclease. AB - We have developed a new electrochemical approach for qualitative and quantitative detection of hepatitis C virus (HCV) based on the site-specific DNA cleavage of BamHI endonuclease. PMID- 19294246 TI - Self-assembly of an M6L12 coordination cube. AB - A 3x3x3 nm cubic coordination compound quantitatively self-assembled from 6 palladium ions and 12 bent ligands with a 90 degrees bend angle. PMID- 19294247 TI - One-pot synthesis of cyclic antifreeze glycopeptides. AB - The first cyclic glycopeptides exhibiting significant antifreeze activity by forming hexagonal-bipyramidal ice crystals, denoted cyclic antifreeze glycopeptides (cyclic AFGPs), were constructed by a one-pot synthesis based on the controlled cyclization reaction of pre-formed small linear glycopeptides. PMID- 19294248 TI - Experimental and theoretical demonstration of ferroelectric anisotropy in a one dimensional copper(II)-based coordination polymer. AB - The anisotropy of polarization in a 1D copper(II)-based coordination polymer was investigated experimentally and theoretically for the first time, revealing that the origin of the ferroelectricity and its anisotropic nature are closely related to the coordination geometry of the metal ion and the packing mode of the coordination polymer. PMID- 19294249 TI - Construction of and electrophoretic fluorescence imaging with a nano-porous polyaniline-fluorescein conducting particle system. AB - Electrophoretic fluorescence images have been fabricated by deposition of nanoporous polyaniline-fluorescein conducting particles onto a micro-patterned polyaniline film. PMID- 19294250 TI - Helicity induction on water-soluble oligoresorcinols in alkaline water and their application to chirality sensing. AB - Oligoresorcinol oligomers were found to adopt a single-stranded helical conformation in alkaline water through intramolecular hydrogen bonds, of which a helix-sense bias was induced by the addition of water-soluble chiral compounds, such as ammonium salts, amines and alcohols. PMID- 19294251 TI - Facile fabrication of triple-layer (Au@Ag)@polypyrrole core-shell and (Au@H2O)@polypyrrole yolk-shell nanostructures. AB - Triple-layer (Au@Ag)@polypyrrole core-shell nanoparticles are fabricated by a one step synthesis involving simultaneous reduction of AgNO3 and polymerization of pyrrole in the presence of Au nanoparticles; the Ag layer in the resulting nanoparticles is etched to give a (Au@H2O)@polypyrrole yolk-shell structure. PMID- 19294252 TI - 'Clicked' magnetic nanohybrids with a soft polymer interlayer. AB - We have developed a facile and efficient methodology to prepare magnetic nanohybrids from 'clickable' magnetic nanoparticles and polymer-coated nanomaterials by Cu(I)-catalyzed azide-alkyne cycloaddition 'click' chemistry. PMID- 19294253 TI - Colorimetric recognition of DNA intercalators with unmodified gold nanoparticles. AB - A colorimetric approach to recognize the binding mode between DNA and molecules was developed, which was based upon the electrostatic interaction between DNA and gold colloids. PMID- 19294254 TI - Discrete MnAlPO-5 nanocrystals synthesized by an ionothermal approach. AB - Discrete MnAlPO-5 nanocrystals with monomodal particle size distribution (diameter of 80 nm) were synthesized using an ionothermal technique; the pore filling effect of the ionic liquid molecules during the synthesis of microporous nanocrystals is verified. PMID- 19294255 TI - Dynamic structural changes of pentacopper(II) chains supported by N6-donor ligands. AB - Elastic pentacopper molecular chains, [Cu5(panapy)4X2] (X=Cl (), Br ()) and [Cu5(panapy)4]X'2 (X'=BF4 (), PF6 ()), were prepared using a naphthyridine modulated N6-donor ligand, panapy2-, and showed magnetically coupled, dynamic rearrangement of five Cu(II) ions switched by the presence/absence of halide termination. PMID- 19294256 TI - Non-covalent functionalization of graphene sheets by sulfonated polyaniline. AB - Graphene sheets were stably dispersed in water by functionalization with sulfonated polyaniline (SPANI), and the composite film of SPANI-functionalized graphene showed improved electrochemical stability and enhanced electrocatalytic activity. PMID- 19294257 TI - A small cationic donor-acceptor iridium complex with a long-lived charge separated state. AB - A cationic cyclometalated iridium complex with a triarylamine donor attached to the primary ligand sphere showed long-lived (0.04 and 1.7 micros) charge separated states after photoexcitation which are due to a combined Marcus inverted region effect and spin-selection rules. PMID- 19294258 TI - BODIPY derivatives as donor materials for bulk heterojunction solar cells. AB - BODIPY derivatives have been used as donor in solution-processed bulk heterojunction solar cells using PCBM as acceptor. A power conversion efficiency of 1.34% has been obtained under simulated solar irradiation. PMID- 19294259 TI - Inhibition of photobleaching and blue shift in quantum dots. AB - Photobleaching and spectral diffusion (blue shift) of quantum dots at the solid/liquid interface are suppressed by adding mercaptoethylamine. PMID- 19294260 TI - Synthesis and structure of air-stable Lewis acidic binuclear complex of zirconocene pentafluorophenylsulfonate and its catalytic application in the allylation of carbonyl compounds with tetraallyltin. AB - Air-stable Lewis acidic micro2-hydroxy bridged binuclear complex of zirconocene pentafluorophenylsulfonate was successfully synthesized and found to show high catalytic efficiency in chemoselective allylation of carbonyl compounds with tetraallyltin in aqueous methanol media. PMID- 19294261 TI - Amine-triggered molecular capsules using dynamic boronate esterification. AB - Boron-contained molecular capsules triggered by amines are described for the first time; when Et3N was employed as a chemical stimulus the Lewis acid Et3NH+, produced by solvolysis reaction on the boron, was found to be encapsulated in the internal cavity, whereas use of nBu3N led to the related dynamic capsule capable of accommodating several guest species including Cs+. PMID- 19294262 TI - Ionic strength-dependent structural transition of proteins at electrode surfaces. AB - Using constant current chronopotentiometry we showed that in 50 mM sodium phosphate (pH 7) bovine serum albumin and some other proteins were not significantly denatured at a bare mercury electrode while at higher phosphate concentrations they underwent electric field-driven denaturation on the electrode surface. PMID- 19294263 TI - Propagation of polymer nanosheets from silica opal membrane gaps by thermal polymerization of bicontinuous microemulsions. AB - Unique polymer morphologies such as fractal surfaces consisting of closely-spaced nanosheets and vertically extended nanosheets were found to be formed by thermal polymerization of styrene or methyl methacrylate bicontinuous microemulsions (BME) from the gaps of a silica opal membrane. PMID- 19294264 TI - Formation of a new class of 7pi radicals via sterically induced P-P bond cleavage of the dimers [(CH)2(NR)2P]2. AB - The 2c-2e- P-P bonded dimers [(CH)2(NR)2P]2 dissociate in solution to give the persistent new 7pi radicals [(CH)2(NR)2P]*, which are isoelectronic with the well known S/N thiazolyl radicals. PMID- 19294265 TI - Photoinduced hole transfer forming polymer quantum dot nanocomposites. AB - The study of hole transfer from CdSe QD core to ligands under visible light irradiation and conductive polymer-QD composites formation without conventional photoinitiator. PMID- 19294266 TI - Multicolour self-assembled particles of fluorene-based bolaamphiphiles. AB - Bolaamphiphilic fluorene-based oligomers self-assemble in water to form fluorescent nanoparticles with tuneable emission colours covering the entire visible range, even including white. PMID- 19294267 TI - A dual-ion-switched molecular brake based on ferrocene. AB - A new dual-ion-switched molecular brake was synthesized and operated by chemical reagents (H+/OH- and Zn2+/EDTA), and its rotation could be read out by chromogenic, fluorogenic and electrochemical signals. PMID- 19294268 TI - Oxovanadium(IV)-based near-IR PDT agents: design to biological evaluation. AB - An oxovanadium(IV) complex of dipyridophenazine, as a potent metal-based PDT agent, shows efficient DNA photocleavage activity at near-IR region and high photocytotoxicity in both UV-A and visible light in HeLa cells. PMID- 19294269 TI - Quantitative formation of [4]pseudorotaxanes from two rods and two bis macrocycles incorporating porphyrinic plates between the rings. AB - [4]pseudorotaxanes consisting of two very large coordinating bis-macrocycles and rigid rods incorporating two side-by-side chelates have been obtained quantitatively utilising the gathering and threading effect of copper(I); the assemblies obtained are several nanometres long and they contain two face-to-face zinc porphyrins which will be used to complex various organic substrates. PMID- 19294270 TI - Selective glucose recognition by boronic acid azoprobe/gamma-cyclodextrin complexes in water. AB - The phenylboronic acid azoprobe (BA-Azo)/gamma-cyclodextrin (gamma-CD) complex exhibits a selective response for D-glucose by forming a supramolecular 2:1 inclusion complex of the azoprobes with D-glucose inside the gamma-CD cavity. PMID- 19294271 TI - Light-driven altitudinal molecular motors on surfaces. AB - A Cu(I)-catalyzed 1,3-dipolar cycloaddition was used to construct a monolayer of an altitudinal molecular motor on quartz and silicon substrates, which represents the fastest light-driven molecular motor, to date, grafted to a solid surface. PMID- 19294272 TI - Copper catalyzed coupling of aryl chlorides, bromides and iodides with amines and amides. AB - A copper(I) catalyzed procedure for carbon-nitrogen bond formation utilizing aryl halides and either amines, including amino acids and diphenylamine, or aliphatic and aromatic amides is described. PMID- 19294273 TI - Electrochemical polymer reaction: selective fluorination of a poly(fluorene) derivative. AB - Anodic fluorodesulfurization of a poly(fluorene) derivative coated on a Pt electrode was successfully carried out in an ionic liquid hydrogen fluoride salt, yielding a alternating copolymer containing a difluorofluorene moiety. PMID- 19294274 TI - Substrate channelling in an engineered bifunctional aldolase/kinase enzyme confers catalytic advantage for C-C bond formation. AB - A new bifunctional enzyme that displays both aldolase and kinase activities has been designed and successfully used in the synthesis of aldol adducts, employing DHA as initial donor, with an increase in the reaction rate of 20-fold over the parent enzymes, which can be interpreted in terms of substrate channelling. PMID- 19294275 TI - A robust procedure for the functionalization of gold nanorods and noble metal nanoparticles. AB - Towards a robust and universal functionalization procedure with alkanethiols for gold nanorods and plasmonic nanoparticles, a straightforward two-step approach is described. PMID- 19294276 TI - Self-assembling optically pure Fe(A-B)3 chelates. AB - Optically pure, single diastereomer fac-tris(diimine) complexes of Fe(II) are available from a remarkably facile one-pot procedure using a range of readily available (R)-2-phenylglycinol derivatives. PMID- 19294277 TI - Versatile, efficient derivatization of polysiloxanes via click technology. AB - Copper-catalyzed (click chemistry) or thermal 1,3-dipolar cycloadditions of molecular or polymeric azido-siloxanes with a broad range of alkynes (such as amino acids or carbohydrates) easily yield new conjugates that would be otherwise difficult, if not impossible, to prepare by traditional methodologies. PMID- 19294278 TI - Chemoenzymatic syntheses of carbasugar analogues of nucleoside diphosphate sugars: UDP-carba-Gal, UDP-carba-GlcNAc, UDP-carba-Glc, and GDP-carba-Man. AB - Chemoenzymatic syntheses of several NDP-carba-sugars have been successfully carried out, and these essential cofactor analogues are expected to be selective inhibitors of glycosyltransferase enzymes. PMID- 19294279 TI - Fluorescent gold nanoparticles-based fluorescence sensor for Cu2+ ions. AB - A new fluorescence sensor for the highly selective detection of Cu2+ ion with a detection limit of 3.6 nM based on the aggregation-induced fluorescence quenching of the highly fluorescent glutathione-capped gold nanoparticles is reported. PMID- 19294280 TI - Oxoammonium salt/NaClO2: an expedient, catalytic system for one-pot oxidation of primary alcohols to carboxylic acids with broad substrate applicability. AB - A facile, green, one-pot oxidation of primary alcohols to carboxylic acids with broad substrate applicability has been developed by employing an expedient catalytic system consisting of 1-Me-AZADO+X-/NaClO2. PMID- 19294281 TI - Fabrication of lanthanide oxide microspheres and hollow spheres by thermolysis of pre-molding lanthanide coordination compounds. AB - A series of lanthanide oxide microspheres and hollow spheres have been fabricated by thermolysis of corresponding lanthanide coordination compounds formed via bottom-up self-assembly. PMID- 19294282 TI - 1,2,3-triphosphole derivatives as reactive intermediates. AB - The complexes [Cp*E{W(CO)5}2] (E=P, As; Cp*=C5Me5) react with 1,2 diphosphinobenzene to give 1,2,3-triphosphole and 2-arsa-1,3-diphosphole derivatives as reactive intermediates that dimerise to polycyclic phosphorous and arsenic containing cage compounds. PMID- 19294283 TI - Functionalization of hexakis methanofullerene malonate crown-ethers: promising octahedral building blocks for molecular networks. AB - Hexaazido and hexaiodo macrocyclic methanofullerenes undergo high-yielding sixfold click reactions or sixfold Heck, Sonogashira and Suzuki cross-coupling reactions, respectively, to yield all organic building blocks for higher molecular architectures. PMID- 19294284 TI - Improved organic hydrogen carriers with superior thermodynamic properties. AB - By the incorporation of N atoms into naphthalene, we present a theoretical investigation to seek for improved organic hydrogen carriers with an explicit guideline, the release of H2 is found to be greatly favored thermodynamically and the corresponding cycloalkanes possess high hydrogen storage capacity, this offers extensive candidates for practical applications of the promising hydrogen energy. PMID- 19294285 TI - Characterization of silver ions adsorbed on gold nanorods: surface analysis by using surface-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-MS) indicated AgBr2-, which adsorbed on gold nanorod surfaces, was a key material to control the anisotropic growth of gold nanorods. PMID- 19294286 TI - Reductive quenching of pyridine linked porphyrins by phenol: a case of proton coupled electron transfer. AB - PCET is reported for free-base meso-(pyridyl)porphyrins, a pure organic system in dichloromethane solution, by employing UV/Vis and fluorescence spectroscopic techniques. PMID- 19294287 TI - Catalytic nitrene transfer from an imidoiron(III) complex to form carbodiimides and isocyanates. AB - The metastable iron(III) imido species LtBuFeNAd catalyzes transfer of the nitrene fragment NAd from an organic azide to isocyanides or CO, forming unsymmetrical carbodiimides or isocyanates. PMID- 19294288 TI - Nanometre-sized titanium dioxide-catalyzed reactions of nitric oxide with aliphatic cyclic and aromatic amines. AB - Activated on the surface of nanometre-sized TiO2, NO gas can easily react with aliphatic cyclic amines and aryl free radicals at RT under atmospheric pressure to offer NONOates and cupferron sodiums, respectively. PMID- 19294289 TI - Conveniently synthesized isophorone dyes for high efficiency dye-sensitized solar cells: tuning photovoltaic performance by structural modification of donor group in donor-pi-acceptor system. AB - A novel isophorone sensitizer D-3 based on a donor-pi-acceptor system has been conveniently synthesized for highly efficient dye-sensitized solar cells, resulting in a remarkable overall conversion efficiency of 7.41% (AM 1.5, 100 mW cm(-2)) with Jsc of 18.63 mA cm(-2), Voc of 634 mV and FF of 0.63. PMID- 19294290 TI - A facile access to [60]fullerene-fused delta-lactones: unexpected reaction pathway of benzenediazonium-2-carboxylates controlled by organic bases. AB - The reaction of [60]fullerene with anthranilic acids and isoamyl nitrite in the presence of triethylamine unexpectedly afforded [60]fullerene-fused delta lactones, which would be difficult to be prepared by other known methods; the organic base played a crucial role in the unusual reaction pathway. PMID- 19294291 TI - C-H oxidation by hydroxo manganese(v) porphyrins: a DFT study. AB - A DFT study on toluene oxidation by Mn(V) porphyrins shows that both MnV(O) and, remarkably, MnV(OH) are potent H-atom abstractors. PMID- 19294292 TI - Formation of Au nanoparticles in an ionic liquid by electron beam irradiation. AB - A novel method to synthesize Au nanoparticles via a reductive reaction in an ionic liquid containing Au3+ ions was demonstrated using a low-energy electron beam irradiation technique; Au nanoparticles (approximately 122 nm) formed by the incident electron beam were well dispersed and crystallized; this finding opens up the possibility that the use of electron beams and ionic liquids is of key importance in the development of new fabrication techniques for nanomaterials. PMID- 19294294 TI - Lock release lithography for 3D and composite microparticles. AB - We present a method called "Lock Release Lithography (LRL)" that utilizes a combination of channel topography, mask design, and pressure-induced channel deformation to form and release particles in a cycled fashion. This technique provides a means for the high-throughput production of particles with complex 3D morphologies and composite particles with spatially configurable chemistries. In this work, we demonstrate a diverse set of functional particles including those displaying heterogeneous swelling characteristics and containing functional entities such as nucleic acids, proteins and beads. PMID- 19294295 TI - Spontaneous, oscillatory liquid transport in surface tension-confined microfluidics. AB - A unique, spontaneous, oscillatory flow phenomenon driven by the Marangoni effect on a heterogeneous surface can be used for passive microfluidic pumping. PMID- 19294296 TI - A new USP Class VI-compliant substrate for manufacturing disposable microfluidic devices. AB - As microfluidic systems transition from research tools to disposable clinical diagnostic devices, new substrate materials are needed to meet both the regulatory requirement as well as the economics of disposable devices. This paper introduces a UV-curable polyurethane-methacrylate (PUMA) substrate that has been qualified for medical use and meets all of the challenges of manufacturing microfluidic devices. PUMA is optically transparent, biocompatible, and exhibits high electroosmotic mobility without surface modification. We report two production processes that are compatible with the existing methods of rapid prototyping and present characterizations of the resultant PUMA microfluidic devices. PMID- 19294297 TI - Use of photopatterned porous polymer monoliths as passive micromixers to enhance mixing efficiency for on-chip labeling reactions. AB - We introduce a passive micromixer with novel architecture using photopatterned porous polymer monoliths (PPM) and demonstrate an improvement in mixing efficiency by monitoring the fluorescence of an on-chip labeling reaction. UV light was used to photopattern a periodic arrangement of PPM structures directly within the channel of a plastic microfluidic chip. By optimizing the composition of the polymerization solution and irradiation time we demonstrate the ability to photopattern PPM in regularly repeating 100 microm segments at the tee-junction of the disposable device. To evaluate the efficiency of this dual functional mixer-reactor fluorescamine and lysine were introduced in separate channels upstream of the tee-junction and the intensity of laser-induced fluorescence resulting from the fluorogenic labeling reaction was monitored. The fluorescence level after the photopatterned periodic monolith configuration was 22% greater than both an equivalent 1 cm continuous segment of PPM and an open channel. Results indicate that this periodic arrangement of PPM, with regularly spaced open areas between 100 microm plugs of PPM, is directly responsible for enhancing the mixing and overall rate of chemical reaction in the system. In addition to facilitating preparation of a dual functional mixer-reactor, the ability to accurately photopattern PPM is an enabling technology for seamlessly integrating multiple monoliths into a single device. This technology will be particularly important to proteomic applications requiring preconcentration, enzymatic digestion and two-dimensional separations. PMID- 19294298 TI - Label-free quantification of asymmetric cancer-cell filopodium activities in a multi-gradient chip. AB - We use novel super-resolution bright-field optical microscopy to observe the filopodium activities of human lung cancer cells in a multi-gradient cell culture chip. Temporal variations of the filopodium numbers are measured without fluorescent labelling. By carefully designing the fluidic field inside the culture chip, we establish stable concentration gradients of the injected reagents. The reagents are injected via a separated central inlet, and the concentration gradients are different at different positions in the chip. The same chip can be used for both control and treated experiments. Using epidermal growth factor as the treatment, we verify that the protrusions of filopodia indicate the direction of concentration gradients experienced by a living cancer cell; while the treatment of bovine serum albumin shows no specific effect on the growth of filopodia. The combination of label-free, high-resolution optical microscopy and a micro cell culture chip establishes a convenient and versatile platform for dynamical cancer-cell analyses. PMID- 19294299 TI - The BAD project: data mining, database and prediction of protein adsorption on surfaces. AB - Protein adsorption at solid-liquid interfaces is critical to many applications, including biomaterials, protein microarrays and lab-on-a-chip devices. Despite this general interest, and a large amount of research in the last half a century, protein adsorption cannot be predicted with an engineering level, design orientated accuracy. Here we describe a Biomolecular Adsorption Database (BAD), freely available online, which archives the published protein adsorption data. Piecewise linear regression with breakpoint applied to the data in the BAD suggests that the input variables to protein adsorption, i.e., protein concentration in solution; protein descriptors derived from primary structure (number of residues, global protein hydrophobicity and range of amino acid hydrophobicity, isoelectric point); surface descriptors (contact angle); and fluid environment descriptors (pH, ionic strength), correlate well with the output variable-the protein concentration on the surface. Furthermore, neural network analysis revealed that the size of the BAD makes it sufficiently representative, with a neural network-based predictive error of 5% or less. Interestingly, a consistently better fit is obtained if the BAD is divided in two separate sub-sets representing protein adsorption on hydrophilic and hydrophobic surfaces, respectively. Based on these findings, selected entries from the BAD have been used to construct neural network-based estimation routines, which predict the amount of adsorbed protein, the thickness of the adsorbed layer and the surface tension of the protein-covered surface. While the BAD is of general interest, the prediction of the thickness and the surface tension of the protein covered layers are of particular relevance to the design of microfluidics devices. PMID- 19294300 TI - DEP actuated nanoliter droplet dispensing using feedback control. AB - Dielectrophoretic (DEP) droplet dispensing using dielectric-coated coplanar electrode structures provides an ideal platform for testing smart control systems for high-speed microfluidic devices. Open-loop control of DEP droplet dispensing is not sufficiently robust for precision droplet dispensing because unavoidable surface property variations of the substrates and other parameters such as liquid viscosity introduce uncertainty in the motion. Closed-loop systems employing distributed optical sensors and feedback provide flexibility, sensitivity, and reliability. In this new scheme, an array of distributed optical sensors detects fluid motion and, through a programmable control module, triggers application of AC voltage bursts of appropriate magnitude, duration, and frequency to control liquid motion and droplet formation. Reconfiguring the module connections and reprogramming the control module permits testing of a variety of control strategies. PMID- 19294301 TI - A novel electrolytic ignition monopropellant microthruster based on low temperature co-fired ceramic tape technology. AB - A planar 2-D liquid monopropellant microthruster fabricated from low temperature co-fired ceramic tapes and ignited by electrolysis is reported. The volume of the combustion chamber was 820 nL (0.82 mm(3)). Silver electrodes were screen printed and positioned on the top and bottom surfaces of the combustion chamber. A DC voltage potential applied across the electrodes was used to initiate decomposition of hydroxylammonium nitrate (HAN) based liquid monopropellants. A thrust output of 150 mN was obtained using a voltage input of 45 V. Measured ignition energies were as small as 1.9 J. Ignition delays, as short as a few hundred milliseconds, were found dependent on the type of HAN-based propellant and the voltage potential. PMID- 19294302 TI - T7-based linear amplification of low concentration mRNA samples using beads and microfluidics for global gene expression measurements. AB - We have demonstrated in vitro transcription (IVT) of cDNA sequences from purified Jurkat T-cell mRNA immobilized on microfluidic packed beds down to single-cell quantities. The microfluidically amplified antisense-RNA (aRNA) was nearly identical in length and quantity compared with benchtop reactions using the same starting sample quantities. Microarrays were used to characterize the number and population of genes in each sample, allowing comparison of the microfluidic and benchtop processes. For both benchtop and microfluidic assays, we measured the expression of approximately 4000 to 9000 genes for sample amounts ranging from 20 pg to 10 ng (2 to 1000 cell equivalents), corresponding to 41 to 93% of the absolute number of genes detected from a 100 ng total RNA control sample. Concordance of genes detected between methods (benchtop vs. microfluidic) and repeats (microfluidic vs. microfluidic) typically exceeded 90%. Validation of microarray by Real-time PCR of a panel of five genes suggests transcription of genes present is approximately six times more efficient with the microfluidic IVT compared with benchtop processing. Microfluidic IVT introduces no bias to the gene expression profile of the sample and provides more efficient transcription of mRNA sequences present at the single-cell level. PMID- 19294303 TI - Jamming prokaryotic cell-to-cell communications in a model biofilm. AB - We report on the physical parameters governing prokaryotic cell-to-cell signaling in a model biofilm. The model biofilm is comprised of bacteria that are genetically engineered to transmit and receive quorum-sensing (QS) signals. The model is formed using arrays of time-shared, holographic optical traps in conjunction with microfluidics to precisely position bacteria, and then encapsulated within a hydrogel that mimics the extracellular matrix. Using fluorescent protein reporters functionally linked to QS genes, we assay the intercellular signaling. We find that there isn't a single cell density for which QS-regulated genes are induced or repressed. On the contrary, cell-to-cell signaling is largely governed by diffusion, and is acutely sensitive to mass transfer to the surroundings and the cell location. These observations are consistent with the view that QS-signals act simply as a probe measuring mixing, flow, or diffusion in the microenvironment of the cell. PMID- 19294304 TI - The deformation of flexible PDMS microchannels under a pressure driven flow. AB - Poly(dimethylsiloxane) (PDMS) microchannels are commonly used microfluidic structures that have a wide variety of biological testing applications, including the simulation of blood vessels to study the mechanics of vascular disease. In these studies in particular, the deformation of the channel due to the pressure inside is a critical parameter. We describe a method for using fluorescence microscopy to quantify the deformation of such channels under pressure driven flow. Additionally, the relationship between wall thickness and channel deformation is investigated. PDMS microchannels of varying top wall thickness were created using a soft lithography process. A solution of fluorescent dye is pumped through the channels at constant volume flow rates and illuminated. Pressure and fluorescence intensity are measured at five positions along the length of the channel. Fluorescence measurements are then used to determine deformation, using the linear relationship of dye layer thickness and intensity. A linear relationship between pressure and microchannel deformation is measured. Pressure drops and deformations closely correspond to values predicted by the model in most cases. Additionally, measured pressure drops are found to be up to 35% less than the pressure drop in a rigid-walled channel, and channel wall thickness is found to have an increasing effect as the channel wall thickness decreases. PMID- 19294305 TI - Continuous focusing of microparticles using inertial lift force and vorticity via multi-orifice microfluidic channels. AB - We developed a new microfluidic method for focusing microparticles through the combined use of inertial lift forces and turbulent secondary flows generated in a topographically patterned microchannel. The mechanism of particle focusing is based on the hydrodynamic inertial forces exerted on particles migrating along a non-circular microchannel, i.e.tubular pinch effect and wall effect, which induce particle movement away from walls and along a specific lateral position in the microchannel. With the extraordinary geometry of multi-orifice microchannel, an ordered and focused particle distribution was achieved at central or side regions according to a particle Reynolds number (Re(p)) range. The focusing of particles was controlled by the particle Reynolds number, microchannel length, and volume fraction of particles in suspension. This method will be beneficial in particle focusing processes in a microfluidic device since it offers continuous, high throughput performance and simple operation. PMID- 19294307 TI - A fritless, EOF microchip pump for high pressure pumping of aqueous and organic solvents. AB - A fritless, microchip electroosmotic flow (EOF) pump is microfabricated and demonstrated on a planar soda lime glass substrate to be capable of supplying reasonable flow rates under high back pressures, such as that required for micro high pressure liquid chromatography (micro-HPLC). The microchip EOF pump is composed of a densely packed microchannel containing 800 nm silica particles and was capable of generating a maximum pressure > 1000 psi ( approximately 7 MPa) and a maximum flow rate of 282 nL/min (aqueous cyclohexylamino alkyl sulfonate (CHES) buffer, 10 mM, pH 9.0, 200 V/cm). Other pumping fluids, such as CHES buffer-acetonitrile mixture (50%, v/v), CHES buffer-methanol mixture (50%, v/v), and pure acetonitrile were also used in a characterization of pump performance that included determinations of the maximum flow rate, maximum pressure, and resulting flow rate against an applied, downstream back pressure. The flow rate under a 200 psi ( approximately 1.4 MPa) back pressure at an applied electric field strength of 250 V/cm ranged from 285 nL/min for aqueous CHES buffer to 44 nL/min for CHES buffer-acetonitrile mixture (50%, v/v), indicating that this EOF pump will meet the future requirements of a micro-HPLC system. PMID- 19294308 TI - Microfluidic assisted synthesis of multi-functional polycaprolactone microcapsules: incorporation of CdTe quantum dots, Fe3O4 superparamagnetic nanoparticles and tamoxifen anticancer drugs. AB - This paper demonstrates a proof-of-concept approach for encapsulating the anticancer drug tamoxifen, Fe3O4 nanoparticles (NPs) and CdTe quantum dots (QDs) into size-controlled polycaprolactone (PCL) microcapsules utilizing microfluidic emulsification, which combined magnetic targeting, fluorescence imaging and drug controlled release properties into one drug delivery system. Cross-linking the composite PCL microcapsules with poly(vinyl alcohol) (PVA) tailored their size, morphology, optical and magnetic properties and drug release behaviors. The flow conditions of the two immiscible solutions were adjusted in order to successfully generate various sizes of polymer droplets. The result showed superparamagnetic and fluorescent properties, and was used as a controlled drug release vehicle. The composite magnetic and fluorescent PCL microcapsules are potential candidates for a smart drug delivery system. PMID- 19294306 TI - A general microchip surface modification approach using a spin-coated polymer resist film doped with hydroxypropyl cellulose. AB - We have developed a simple and effective method for surface modification of polymer microchips by entrapping hydroxypropyl cellulose (HPC) in a spin-coated thin film on the surface. Poly(methyl methacrylate-8.5-methacrylic acid), a widely available commercial resist formulation, was utilized as a matrix for dissolving HPC and providing adherence to native polymer surfaces. Various amounts of HPC (0.1-2.0%) dissolved in the copolymer and spun on polymer surfaces were evaluated. The modified surfaces were characterized by contact angle measurement, X-ray photoelectron spectroscopy and atomic force microscopy. The developed method was applied on both poly(methyl methacrylate) and cyclic olefin copolymer microchips. A fluorescently labeled myoglobin digest, binary protein mixture, and human serum sample were all separated in these surface-modified polymer microdevices. Our work exhibits an easy and reliable way to achieve favorable biomolecular separation performance in polymer microchips. PMID- 19294309 TI - Microchip-based homogeneous immunoassay using fluorescence polarization spectroscopy. AB - We have realized fluorescence polarization immunoassay (FPIA) on a microchip in about 1 minute. FPIA is a homogeneous competitive immunoassay which is based on measuring fluorescence polarization after competitive binding of an analyte and a tracer to an antibody. We constructed a microfluidic FPIA system composed of a newly designed microchip, a laser, a CCD camera and an optical microscope with two specially installed polarizers-one fixed and one rotatable. Theophylline, a typical small drug molecule, was used as a model analyte. Theophylline and fluorescence-labeled theophylline were introduced through different inlets and combined in a 100 microm-wide microchannel where anti-theophylline antibody was added. To optimize the microchip design for FPIA, we investigated the diffusion time of theophylline and the mixing time of theophylline and antibody in this channel, which were 6 s and 36 s, respectively. We successfully carried out a quantitative analysis of theophylline in serum near the therapeutic range in 65 s. In FPIA, a larger tracer-antibody complex emits more polarized fluorescence than the tracer, and therefore, by increasing the antigen concentration in a sample, more polarization relaxation is observed since the tracer-antibody complex concentration is decreased and the tracer concentration is increased. Tracer binding to an antibody is directly measured by spectroscopic techniques without any separation process.This microchip-based FPIA is very simple and rapid, unlike microchip-based heterogeneous immunoassay, because it does not require several processes such as washing and reflowing and immobilizing of antibodies or antigens in the channel. In the future, microchip-based FPIA should find frequent use for point-of-care testing in the clinical field, where conventional FPIA has been used for laboratory tests. PMID- 19294310 TI - Microfluidic structures for flow cytometric analysis of hydrodynamically focussed blood cells fabricated by ultraprecision micromachining. AB - We present three-dimensional microfluidic structures with integrated optical fibers, mirrors and electrodes for flow cytometric analysis of blood cells. Ultraprecision milling technique was used to fabricate different flow cells featuring single-stage and two-stage cascaded hydrodynamic focusing of particles by a sheath flow. Two dimensional focussing of the sample fluid was proven by fluorescence imaging in horizontal and vertical directions and found to agree satisfactorily with finite element calculations. Focussing of the sample stream down to 5 microm at a particle velocity of 3 m s(-1) is accessible while maintaining stable operation for sample flow rates of up to 20 microL min(-1). In addition to fluorescence imaging, the micro-flow cells were characterised by measurements of pulse shapes and pulse height distributions of monodisperse microspheres. We demonstrated practical use of the microstructures for cell differentiation employing light scatter to distinguish platelets and red blood cells. Furthermore, T-helper lymphocytes labelled by monoclonal antibodies were identified by measuring side scatter and fluorescence. PMID- 19294311 TI - Hydrodynamic resistance of single confined moving drops in rectangular microchannels. AB - We integrate a sensitive microfluidic comparator into a T-junction device and report measurements of the excess pressure drop due to a single moving droplet confined in a rectangular microchannel. We specifically focus on drops that are not coated with surfactants and study the effects of drop size, droplet viscosity and capillary number on their hydrodynamic resistance. In the capillary number range of approximately 10(-3)-10(-2), we find two distinct regimes for hydrodynamic resistance behavior based on drop size. In regime I associated with small drops (drop length/channel width approximately <4), we find that the pressure drop is independent of the drop size and the capillary number, and depends weakly on the ratio of the viscosities of the two immiscible phases. In regime II, associated with large drops (drop length/channel width > approximately 4), depending on the viscosity ratio of the two phases, the hydrodynamic resistance could increase, decrease or remain unchanged with drop size. We present a simple model that qualitatively captures these experimental trends. This model reveals that the pressure drop in regime I is dominated by the dissipation due to the end caps, and in regime II by both the end caps and the central body of the droplet. Such fundamental understanding will enable the design of large-scale energy-efficient fluidic circuits by minimizing the overall pressure drop in a network and may also provide insights into controlling droplet traffic to build functional passively-driven two-phase microfluidic technologies. PMID- 19294312 TI - A micro-ELISA system for the rapid and sensitive measurement of total and specific immunoglobulin E and clinical application to allergy diagnosis. AB - We have developed a novel, practical micro-ELISA system for sensitive and rapid allergy diagnosis. The enzymatic reactions occurred under stopped-flow conditions, resulting in both high precision and high sensitivity. A BSA-biotin avidin linker was introduced for the immobilization of water-soluble allergens on polystyrene microbeads, enabling immobilization of allergens in sufficient density to provide high sensitivity. Evaluation of the system's performance showed a good detection limit (2 ng/mL) for total IgE measurement. In addition, a good correlation with a conventional method (CAP method) was demonstrated using human serum samples from 85 allergy patients. Importantly, sample volumes (5 microL) were 10 times smaller and analysis time (12 min) was >20 times faster than the conventional method. All procedures were automatically regulated with our simple microfluidic system, and all the fluidic, optic and electronic components were integrated for portability. We believe that our system has the potential to become a very powerful tool, particularly for point-of-care diagnosis. PMID- 19294313 TI - Generation of alginate microfibers with a roller-assisted microfluidic system. AB - This work describes a convenient microfluidic strategy with a simple and minimal requirement design for the generation of uniformly-sized alginate gel fibers with diameters one order of magnitude smaller than those formed in conventional methods. Different from common microfluidic methods, our system contains a single microchannel through which alginate solutions are transported and squeezed into a calcium chloride solution. Ca2+ ions immediately cross link the alginate solution into a gel state that is collected with a rotor. We use microchannels to define the initial size of the alginate fibers and a roller to further reduce the fiber size (diameter) by one order of magnitude down to approximately 1 microm. The size of the formed fibers can be well controlled by adjusting the corresponding parameters, and be predicted with a simple equation. In addition, various functional materials (colloidal particles, bacteria and nanoparticles) have been encapsulated into the gel fibers with this technique. PMID- 19294314 TI - The effect of adhesion promoter on the adhesion of PDMS to different substrate materials. AB - The effect of the adhesion promoter GE SS4120 on the adhesion strength of PDMS to different substrates was tested. The adhesion to silicon, glass and aluminium was significantly increased, while adhesion of PDMS to Teflon remained poor, and the adhesion strength of PDMS to PDMS decreased. PMID- 19294315 TI - Multiplex transport and detection of cytokines using kinesin-driven molecular shuttles. AB - The application of biomolecular active transport systems offers a potential route for downscaling multiple analyte assays for lab-on-a-chip applications. Recently, the capture and transport of a wide range of target analytes including proteins, virus particles, and DNA have been demonstrated using kinesin-driven molecular shuttles. The molecular shuttles consisted of microtubule (MT) filaments that were functionalized with either analyte-selective antibodies or complementary DNA, thus facilitating selective target capture and transport. In the present work, we have applied this microfluidic platform for the simultaneous detection of multiple target protein analytes. Multiplexing of molecular shuttles was achieved by immobilizing biotinylated antibodies against interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha) on biotinylated MTs using a streptavidin bridge. Nanocrystal quantum dots of different sizes and spectral emissions were functionalized with IL-2 and TNF-alpha antibodies to facilitate multiplexed detection. In this paper we discuss the results of selectivity and motility in single and multiplexed assays. PMID- 19294316 TI - Microfluidic stickers for cell- and tissue-based assays in microchannels. AB - Difficulties in culturing cells inside microchannels is a major obstacle for the wide use of microfluidic technology in cell biology. Here, we present a simple and versatile method to interface closed microchannels with cellular and multicellular systems. Our approach, based on microfluidic stickers which can adhere to wet glass coverslips, eliminates the need to adapt cell culture conditions to microchannels and greatly facilitates the methods required to position cells into microcircuits. We demonstrate the simplicity and efficiency of the method with HeLa cells, primary cultured neurons and Drosophila tissues. PMID- 19294317 TI - When one chip is not enough: augmenting the validity of SELDI-TOF proteomic profiles of clinical specimens. AB - To improve recovery, selectivity and reproducibility of SELDI-TOF analyses, we found it necessary to modify manufacturer's recommended protocols on sample and chip preparation. To yield reproducible denaturing conditions we verified concentrations of denaturing, reducing and lipid-solubilizing agents. We improved sorption of molecules of interest and reproducibility of analyses by introducing the preconditioning step and alkaline/acidic elutions for normal phase chips. The ratio that reproducibly decomposed the specimen was urea 9 mol l(-1) + DTT 10 mmol l(-1) + CHAPS 20 g l(-1). For sample denaturation, 100 microl of the fresh mixture was added to 100 microl of the specimen. Our modification of a chip processing increased recovery of the NP20 chip by up to 400% as assessed by total ion current. We obtained the range of mass accuracy of 0.02-0.04% and response precision between 30-40% of m/z+. We observed about 50% peak overlap. To obtain approximately 92% of possible peaks three chip selectivities, IMAC, H50 and normal phase with alkaline wash should be used. The selectivity of the SELDI chips is affected by unspecific interactions of a sample with a chip backbone. The system is compatible with matrix-based biological materials and does not suffer from urea interference and sensitivity to covalently bound alkaline ions. The technique is reasonably suitable for semiquantitative screening in the mammalian low-molecular weight cellular, tissue and plasma proteome. PMID- 19294318 TI - Exploring gender, work and living conditions and health - suggestions for contextual and comprehensive approaches. AB - New methods are needed to complement traditional epidemiological methods in the analyses of complex multivariate exposures. Contextual and comprehensive analyses such as multilevel and cluster analyses are particularly suitable for comparisons of women's and men's risk factors for health in working life as well as for guidance in preventive action. Such methodologies are presented in this discussion paper. Contextual and comprehensive (or holistic) approaches help identify the gender-structured situations for women and men and facilitate the discovery of differences among all women and all men. Empirical studies using these approaches show that it is important to take gender composition in both the workplace and the sector into consideration when exploring risk factors for health. PMID- 19294319 TI - Gender adjustment or stratification in discerning upper extremity musculoskeletal disorder risk? AB - OBJECTIVES: The aim was to explore whether "adjustment" for gender masks important exposure differences between men and women in a study of rotator cuff syndrome (RCS) and carpal tunnel syndrome (CTS) and work exposures. METHODS: This cross-sectional study of 733 subjects in 12 health care and manufacturing workplaces used detailed individual health and work exposure assessment methods. Multiple logistic regression analysis was used to compare gender stratified and adjusted models. RESULTS: Prevalence of RCS and CTS among women was 7.1% and 11.3% respectively, and among men 7.8% and 6.4%. In adjusted (gender, age, body mass index) multivariate analyses of RCS and CTS, gender was not statistically significantly different. For RCS, upper arm flexion >/=45 degrees and forceful pinch increased the odds in the gender-adjusted model (OR 2.66, 95% CI 1.26-5.59) but primarily among women in the stratified analysis (OR 6.68, 95% CI 1.81-24.66 versus OR 1.45, 95% CI 0.53-4.00). For CTS, wrist radial/ulnar deviation >/=4% time and lifting >/=4.5kg >3% time, the adjusted OR was higher for women (OR 4.85, 95% CI 2.12-11.11) and in the gender stratified analyses, the odds were increased for both genders (women OR 5.18, 95% CI 1.70-15.81 and men OR 3.63, 95% CI 1.08-12.18). CONCLUSIONS: Gender differences in response to physical work exposures may reflect gender segregation in work and potential differences in pinch and lifting capacity. Reduction in these exposures may reduce prevalence of upper extremity disorders for all workers. PMID- 19294320 TI - A cross-sectional study of mental health and well-being and their associations in the UK veterinary profession. AB - BACKGROUND: Veterinary surgeons are at elevated risk of suicide, with a proportional mortality ratio around four times that of the general population and approximately twice that of other healthcare professions. There has been much speculation regarding possible mechanisms underlying increased suicide risk in the profession but little empirical research. We aimed to assess the contribution of mental health and well-being to the elevated risk, through a postal questionnaire survey of a large stratified random sample of veterinary surgeons practising within the UK. METHODS: A questionnaire was mailed twice to 3,200 veterinary surgeons. Anxiety and depressive symptoms, alcohol consumption, suicidal ideation, positive mental well-being, perceptions of psychosocial work characteristics, and work-home interaction were assessed using valid and reliable existing instruments and a series of bespoke questions previously developed through informal focus groups. RESULTS: Evaluable questionnaires were returned by 1,796 participants, a response rate of 56.1%. The demographic and occupational profile of respondents was representative of the UK veterinary profession. The prevalence of 'caseness' (i.e. HADS subscale score >/=8) for anxiety, depression, and co-morbid anxiety and depression was 26.3, 5.8 and 4.5%. 5.4% of respondents were non-drinkers, 32.0% low-risk drinkers, and 62.6% 'at-risk' drinkers (i.e. AUDIT-C score >/=4 for women, >/=5 for men). The 12-month prevalence of suicidal thoughts was 21.3%. CONCLUSIONS: Compared to the general population, the sample reported high levels of anxiety and depressive symptoms; higher 12-month prevalence of suicidal thoughts; less favourable psychosocial work characteristics, especially in regard to demands and managerial support; lower levels of positive mental well-being; and higher levels of negative work-home interaction. The levels of psychological distress reported suggest ready access to and knowledge of lethal means is probably not operating in isolation to increase suicide risk within the profession. PMID- 19294321 TI - Academic attainment: a predictor of psychiatric disorders? AB - AIM: Poor academic performance can reflect the neurobiological abnormalities underlying schizophrenia and has been suggested as a premorbid marker for the disorder. However, it is not clear as to how academic performance compares as a marker for other mental disorders. The objective of this study was to examine whether educational attainment is a candidate marker for certain psychiatric disorders. METHODS: Using a retrospective cohort design, we compared the early educational performance of 6957 National Service male conscripts. The presence of mental illness was assessed using the Composite International Diagnostic Interview. Academic performance was assessed using the results of a standardized national examination after 6 years of primary education. RESULTS: Adjusting for ethnicity, the scores from this examination revealed that those with schizophrenia spectrum disorders scored significantly lower than those without any mental illness. CONCLUSIONS: Poor educational attainment predicts the onset of schizophrenia spectrum disorders and could be a possible predictor for this specific group of mental illness. PMID- 19294322 TI - Risk of mental disorders in refugees and native Danes: a register-based retrospective cohort study. AB - BACKGROUND: Refugees are vulnerable to mental health disorders because of migration trauma. However, register-based prevalence studies are missing. AIMS: To investigate the risk of mental disorders among refugees compared with that among native Danes. METHOD: Refugees (n = 29,139), who received residence permission in Denmark from 1.1.1993 to 31.12.1999 were matched 1:4 on age and sex with native Danes (n = 116,556). Civil registration numbers were linked to the Danish Psychiatric Central Register to obtain data on ICD-10 diagnosis upon discharge for all first-time psychiatric hospital contacts for refugees (n = 2,120) and native Danes (n = 5,044) between 1.1.1994 and 31.12.2003. Treated prevalence was then calculated using a Poisson regression model. RESULTS: Refugee men (RR = 2.02; 95%CI = 1.75-2.34) and refugee women (RR = 1.49; 95%CI = 1.29 1.72) had higher overall risks of having a first-time psychiatric contact for mental disorders than did native Danes; specific risks of psychotic, affective and neurotic disorders were even higher. The results were most striking for refugee men, and for refugees from the former Yugoslavia, Iraq and the Middle East. CONCLUSIONS: Refugees have high rates of various mental disorders. Healthcare services should target refugees' mental health from arrival in the receiving country. PMID- 19294323 TI - Secondary prevention of hazardous alcohol consumption in psychiatric out patients: a randomised controlled study. AB - BACKGROUND: Hazardous alcohol use is associated with an increased risk for development of a substance use disorder, leading to negative outcomes in psychiatric patients. AIMS: In order to investigate whether psychiatric outpatients' hazardous alcohol consumption could be reduced by way of a brief intervention by telephone. METHOD: Non-psychotic psychiatric outpatients, n = 1,670, completed a self-rating form concerning alcohol habits (AUDIT). Participants with scores indicating risk consumption (n = 344) were randomised to intervention (immediate advice) or control (advice after 6 months). RESULTS: Hazardous alcohol habits occurred among 19% of the women and 24% of the men. In the intervention group, half of the patients reduced their alcohol consumption to non-hazardous levels at 6-month follow-up (ITT analysis). In women, 41.5% in the intervention group had no hazardous consumption at follow-up compared to 24.7% in the control group (P = 0.003), corresponding figure for men was 49.1 and 34.0%. CONCLUSION: Brief intervention seems to be effective to reduce hazardous alcohol consumption in psychiatric outpatients. PMID- 19294324 TI - A community-based study of case fatality proportion among those who carry out suicide acts. AB - OBJECTIVE: To investigate the case fatality proportion and associated factors in those carrying out suicide acts in Nantou, Taiwan. METHOD: Data from 1,171 suicide acts (including 973 with deliberate self harm and 198 completed suicides), identified between July 2000 and February 2003, were collected from a population suicide register in Nantou County, Taiwan. Case fatality proportion and the independent effects of demographic factors and suicide method on case fatality were investigated. RESULTS: The overall case fatality proportion was 16.9%, with higher proportions in men (26.3%) and in those aged 65 and over (37.9%). Hanging was the most lethal method (fatality proportion = 81.5%); pesticide was both commonly used and associated with a high case fatality (fatality proportion = 26.3%). Only age (increased with age) and suicide method were found to be independently associated with the risk of fatality. CONCLUSIONS: Suicide method and older age are independent predictors of the fatality of suicide acts. Suicide prevention strategy and clinical assessment ought to take into account of these two factors. Diminution of pesticide toxicity and control of access to pesticides are important considerations for suicide prevention in rural Taiwan. PMID- 19294325 TI - Mental health service use in a nationwide sample of Korean adults. AB - INTRODUCTION: An understanding of the factors leading to the use of mental health services is important in improving access to mental health-care. The purpose of this study was to assess the use of mental health services, determinants of use and barriers to use. METHODS: Data were derived from a representative sample of the general population, aged 18-64 years. The Korean version of the Composite International Diagnostic Interview was used for the purpose of psychiatric assessment. Respondents were also asked about their use of mental health services, and about any experience of barriers to services. The response rate was 80.2%, and a total of 6,275 persons participated in the study. RESULTS: Of the respondents who completed the interview (n=6,275), 1.9% reported the use of mental health services during the past 12 months. Of the respondents who met the criteria for the 1-year diagnosis of a psychiatric disorder (n=916), 6.1% received mental health-care. Age, unemployment, and medical assistance (public assistance) were correlated positively with frequency of mental health-care. Of those who suffered from a psychiatric disorder but did not seek consultation (n=836), 23.4% said that they desired to handle the problem on their own, and 23% asserted that they had no psychiatric disorder. CONCLUSIONS: The high rate of non consultation among those with psychiatric disorders constitutes an important public health problem. Public health efforts to narrow the gap in service delivery are crucial. Also, additional efforts are warranted to address barriers to mental health care to develop an efficient mental health-care system. PMID- 19294326 TI - Effect of cadmium and salinity stresses on growth and antioxidant enzyme activities of wheat (Triticum aestivum L.). AB - A hydroponics experiment was conducted to investigate the effect of salinity (NaCl) and cadmium (Cd) stresses on growth, lipid peroxidation, and antioxidant enzyme activities of three wheat cultivars differing in salt tolerance. Cd and NaCl stresses inhibited plant growth, reduced chlorophyll content, and increased melondialdehyde content and the activities of superoxide dismutase, catalase and peroxidase. The combined effect of NaCl and Cd on these parameters was larger than both NaCl and Cd alone. There was an obvious difference in the response to the both stresses among the three genotypes, with Pir Sabak-85 being less affected. PMID- 19294327 TI - Organochlorine pesticides residues in bottled drinking water from Mexico City. AB - This work describes concentrations of organochlorine pesticides in bottled drinking water (BDW) in Mexico City. The results of 36 samples (1.5 and 19 L presentations, 18 samples, respectively) showed the presence of seven pesticides (HCH isomers, heptachlor, aldrin, and p,p'-DDE) in bottled water compared with the drinking water standards set by NOM-127-SSA1-1994, EPA, and World Health Organization. The concentrations of the majority of organochlorine pesticides were within drinking water standards (0.01 ng/mL) except for beta-HCH of BW 3, 5, and 6 samples with values of 0.121, 0.136, and 0.192 ng/mL, respectively. It is important monitoring drinking bottled water for protecting human health. PMID- 19294328 TI - Persistent DDE in the Mesopotamian wetlands of southern Iraq. AB - Screening of potential pollutants in surface sediments revealed that almost all persistent organochlorine pesticides were not detected in the newly flooded Mesopotamian wetlands of southern Iraq. This observation suggests that there has been minimal input of organochlorine pesticides recently except for p,p'-DDE which was the only pesticide residue detected (0.29-2.33 microg/kg). It was found in all samples indicating its ability to persist under severe drying of previously exposed surface sediments, high temperature, and intensive solar radiation. p,p'-DDE appears to have a negative relationship with wetland biota, such as zooplankton. PMID- 19294329 TI - Self-reported frailty is associated with low calcaneal bone mineral density in a multiracial population of community-dwelling elderly. AB - INTRODUCTION: Low bone mineral density (BMD) is associated with frailty assessed using performance-based measures. However, the latter can be cumbersome and difficult to standardize. We examined whether an easily obtained self-reported frailty measure also predicted low BMD. METHODS: In 230 elderly (82% female, 58% African-American), calcaneal BMD was measured by DXA and frailty evaluated using the VES-13 questionnaire. In addition to the original scoring, we developed a modified scoring system which provided a broader assessment of frailty and excluded age, which is a known independent predictor of BMD. A telephone interview conducted 6 years later ascertained interval fragility fractures and survival status. RESULTS: A higher modified frailty score was associated with lower BMD (p = 0.002), even after adjusting for age, weight, sex, and race and was more predictive of death at 6 years (p = 0.009) than the original score (p = 0.08). Based on our model, a subject with the highest frailty score differed from an otherwise similar subject with the lowest score by a calcaneal BMD of 1.4 T score units, corresponding to 2-3 times higher fracture risk. CONCLUSION: Self reported frailty is associated with low calcaneal BMD and can be used to identify subjects with a greater risk of osteoporosis than expected from traditional risk factors. PMID- 19294330 TI - Spine surgery training and competence of European neurosurgical trainees. AB - BACKGROUND: Little is known about the nature of spine surgery training received by European neurosurgical trainees during their residency and the level of competence they acquire in dealing with spinal disorders. METHODS: A three-part questionnaire entailing 32 questions was devised and distributed to the neurosurgical trainees attending the EANS (European Association of Neurosurgical Societies) training courses of 2004. RESULTS: Of 126 questionnaires, 32% were returned. The majority of trainees responding to the questionnaire were in their final (6(th)) year of training or had completed their training (60.3% of total). Spinal surgery training in European residency programs has clear strengths in the traditional areas of microsurgical decompression for spinal stenosis and disc herniation (77-90% competence in senior trainees). Deficits are revealed in the management of spinal trauma (34-48% competence in senior trainees) and spinal conditions requiring the use of implants and anterior approaches, with the exception of anterior cervical stabilisation. CONCLUSIONS: European neurosurgical trainees possess incomplete competence in dealing with spinal disorders. EANS trainees advocate the development of a postresidency spine subspecialty training program. PMID- 19294331 TI - Oronasopharyngeal chordomas. AB - BACKGROUND: Chordomas are rare tumors derived from notochordal remnants. The authors report on a series of three cases of primary familial oronasopharyngeal chordomas treated at our institution. METHODS: A retrospective chart review was completed of the three cases of primary familial oronasopharyngeal chordoma treated at the University of Utah. FINDINGS: All three patients (100%) were neurologically intact and presented with nasal obstruction. The patients ranged in age from 5 to 65 years and were first-degree relatives. None of the patients had bony erosion of the skull base on imaging, and all of the patients' tumors connected with the skull base via a tract. All three patients were treated with a wide excision combined with drilling of the involved skull base. They all tolerated the procedure without any complications and remain tumor free with a follow-up of 12 months to 4.5 years. CONCLUSION: Primary oronasopharyngeal chordomas are rare tumors that may present without bony erosion of the skull base. A wide excision with drilling of the involved bony structures may offer an oncologic cure. PMID- 19294332 TI - TNF-related apoptosis-inducing ligand level in Alzheimer's disease. AB - In the present study, we determined the significance of tumor necrosis factor related apoptosis-inducing ligand (TRAIL) in Alzheimer's disease (AD). We characterized the expression of TRAIL protein in the cerebrospinal fluid (CSF) and serum with ELISA and TRAIL mRNA in the peripheral blood mononuclear cells (PBMCs) with real-time PCR in 22 patients with AD and 20 control cases. We could not find TRAIL protein in the CSF samples. The concentration of TRAIL protein in sera from patients with AD was not different from controls. However, there was an inverse correlation between serum TRAIL levels and Mini-Mental State Examination scores in AD patients. Also we did not find significant difference in TRAIL mRNA in the PBMCs of patients with AD when compared with control group. Our data indicate that TRAIL serum level decreases in the late stage of disease. PMID- 19294333 TI - Aortic dissection as a possible cause of pure transient global amnesia: a case report and literature review. AB - A 55-year-old man suddenly developed anterograde and retrograde amnesia. His colleagues witnessed the onset of the episode and reported that 2 h before the onset of the amnesic attack the patient transiently became pale. Physical examination was unremarkable and neurological examination revealed no focal neurological sign although a laboratory investigation revealed leukocytosis. Pure transient global amnesia (TGA) was diagnosed. The anterograde amnesia resolved 20 h after onset, but the causes of his transient paleness precedent to TGA and leukocytosis were unclear. Thirty-four hours after onset, the patient complained of sudden back pain and radiological studies revealed aortic dissection (AD; Stanford type B). We emphasize AD as a rare cause of pure TGA, because TGA in itself often has a benign natural history, but AD can be life-threatening if undiagnosed. The precedent pain, transient systemic symptoms, and leukocytosis can be red flags suggesting AD as an etiology of TGA. PMID- 19294335 TI - Interatrial thrombus-in-transit resulting in paradoxical thromboembolism. AB - Patent foramen ovale (PFO) is reported in up to 50% of patients with cryptogenic stroke. However, the role of PFO in acute myocardial infarction is less reported. In this case report, the relationship between PFO, myocardial infarction, and an interatrial paradoxical thromboembolism (aka thrombus-in-transit) was diagnosed with the use of non-invasive technique, percutaneous procedures, as well as gross surgical specimen. PMID- 19294334 TI - Is there a space-time continuum in olfaction? AB - The coding of olfactory stimuli across a wide range of organisms may rely on fundamentally similar mechanisms in which a complement of specific odorant receptors on olfactory sensory neurons respond differentially to airborne chemicals to initiate the process by which specific odors are perceived. The question that we address in this review is the role of specific neurons in mediating this sensory system--an identity code--relative to the role that temporally specific responses across many neurons play in producing an olfactory perception--a temporal code. While information coded in specific neurons may be converted into a temporal code, it is also possible that temporal codes exist in the absence of response specificity for any particular neuron or subset of neurons. We review the data supporting these ideas, and we discuss the research perspectives that could help to reveal the mechanisms by which odorants become perceptions. PMID- 19294336 TI - Decision-making about inherited cancer risk: exploring dimensions of genetic responsibility. AB - Since genetic information has implications for family members, some choices about genetic risk may be influenced by perceptions of responsibility to relatives. Drawing upon 25 semi-structured interviews with test recipients in Canada, this study explored decisions about inherited breast-ovarian and colon cancer. Qualitative data analysis revealed the pervasive significance of genetic responsibility in test decisions. We highlight three dimensions of genetic responsibility: 1) to know about the self for self; 2) to know about the self for others; 3) to know about the self to oblige others to know. It is argued that these dimensions of genetic responsibility have implications for test decisions, family relationships and other family members' desire to know (or not know) and to act (or not act) with respect to their own genetic risk. In particular, genetic responsibility may play out as a framing of a relative's moral obligation to know their risk that could obviate any interest they might have in not knowing. We conclude that perceptions of responsibility to-and of-other family members be thoroughly explored in genetic counseling sessions. PMID- 19294337 TI - Resistance to cadmium as a function of Caco-2 cell differentiation: role of reactive oxygen species in cadmium- but not zinc-induced adaptation mechanisms. AB - Cadmium (Cd) is a highly toxic metal that enters the food chain. Following oral ingestion, the intestinal epithelium is the first biological barrier crossed by Cd and is also an important target tissue. In the present study, the human intestinal Caco-2 cell line was used to evaluate the impact of a low level of exposure on both undifferentiated and differentiated intestinal cells. As revealed by the LC(50) values estimated with the 3-[4,5-dimethyl-2-thiazol-2-yl] 2,5-diphenyltetrazolium bromide (MTT) assay, mature Caco-2 cells were more resistant to Cd. However, following a 24-h exposure to non-cytotoxic levels of Cd (10 microM) or zinc (Zn, 100 microM), threefold increases were obtained in the LC(50) values of 7-day-old cells, whereas increased resistance in 21-day-old cells was observed exclusively with Zn. Induction of MT-IIa and HSP70 mRNAs was higher in undifferentiated cells and an increase in cellular glutathione (GSH) content was observed exclusively in these cell cultures. However, the results obtained with cycloheximide used for inhibiting protein synthesis and with L buthionine sulfoximine (BSO), which inhibits GSH synthesis, revealed that protein synthesis is not a prerequisite to the development of resistance. The presence of 100 mM 3-amino-1,2,4-triazole (3AT), a catalase inhibitor, prevented Cd-induced but not Zn-induced resistance, as well as sensitized cells to Cd toxicity. These results show for the first time differences in constitutive and acquired resistance to Cd as a function of enterocytic differentiation status and suggest the involvement of different mechanisms for Cd- and Zn-induced adaptation in the intestinal cells. Redox signals may trigger Cd-induced adaptation mechanisms but pro-oxidant conditions would eliminate proliferative intestinal cells capability to develop resistance. This would be critical for Cd- but not Zn-induced mechanisms of resistance since Cd but not Zn may cause oxidative stress. PMID- 19294338 TI - [Actives. A central component of modern light protection]. AB - Dermatologists should endorse UV filters as effective and useful. In recent years, numerous technological advances have made it possible to protect the skin better than ever before against the negative effects of both UVAB and UVA radiation. On the other hand, the protection is never 100%. Thus it is wise to combine UV filters with other protective measures. Molecules used for this purpose differ from UV filters in that they cannot absorb or reflect UV radiation. Instead, they damper or block the secondary biochemical processes that follow absorption of UV radiation. The best example for this group is antioxidants. Currently other new active cosmetic ingredients or actives are being developed that can reduce UV-induced skin damage in a variety of ways. PMID- 19294339 TI - [10 frequently asked questions about magnetic resonance imaging in patients with metal implants]. PMID- 19294341 TI - Penicillium marneffei SKN7, a novel gene, could complement the hypersensitivity of S. cerevisiae skn7 Disruptant strain to oxidative stress. AB - Penicillium marneffei is an intracellular fungal pathogen. Being resistant to the reactive oxygen species (ROS) produced by phagocytes is a key step to the surviving of P. marneffei within the phagocytes, as well as its invasion to the host. SKN7, a transcription factor, contributes to the oxidative stress response in yeasts. We cloned the SKN7 ortholog in P. marneffei and compared its transcription on both mycelial and yeast phase, identified its function by complement the S. cerevisiae skn7 disruptant strain. The result showed the full sequence of SKN7 gene was about 2.5 kb, open reading frame extended to 1,845 bp and encoded a putative protein of 614 amino acids. Comparative analysis of the nucleotide sequence of genomic DNA and cDNA confirmed the presence of four introns and two highly conserved HSF-DNA-bind and REC domain. The deduced amino acid sequence was homologous to SKN7 from other fungi. Further, P. marneffei cDNA can partly complement S. cerevisiae skn7 disruptant strain, which was not viable in the presence of 2.5 mM H2O2. The expression level of SKN7 on the two phases, however, had no difference. These results indicated that P. marneffei SKN7 could response to oxidative stress and it was not a phase-specific gene. PMID- 19294342 TI - Biological and microcalorimetric studies of the toxic effect of organoarsenic(V) compounds to wild strain of Bacillus thuringiensis. AB - Microcalorimetric and biological methods were carried out to determine the toxicity of dimethylarsinic acid (DMA) and monomethylarsonic acid (MMA) to wild strain of Bacillus thuringiensis. Thermokinetic parameters were obtained from the power-time curves, showing that the peak-heat output power, total heat output, and number of colonies decreased with the increases in concentration of DMA and MMA. In addition, the generation time and peak maximal time increased with the increases in the dosage of DMA and MMA. The half inhibitory concentrations of DMA and MMA were 99.02 and 142.02 microg mL(-1), respectively for the wild strain of B. thuringiensis. DMA shows higher toxicity to bacteria than MMA. The toxicity resistance of B. thuringiensis against organoarsenic(V) is quite high for the wild strain. Our work demonstrates that microcalorimetry is a very sensitive, simple, and useful technique for in vitro investigation of the toxic effect of organoarsenic(V) on microbial activity. PMID- 19294340 TI - SAS weekly rounds: avascular necrosis. AB - Osteonecrosis of the femoral head is a condition that affects upwards of 10,000 individuals in the USA each year. The peak incidence is in the fourth decade of life, and overall, there is a male preponderance. The condition accounts for up to 12% of total hip arthroplasties performed in developed countries. The etiology can be traumatic or non-traumatic, with 90% of atraumatic cases attributed to corticosteroid therapy or excess alcohol consumption. Osteonecrosis of the femoral head reflects the final common pathway of a range of insults to the blood supply and ultimately results in femoral head collapse, acetabular involvement, and secondary osteoarthritis. Currently, conservative treatment options, which aim to correct pathophysiologic features allowing revascularization and new bone formation, appear to be able to delay but not halt the progression of this condition. As a consequence of femoral head osteonecrosis, many individuals undergo surgical treatments including: core decompression, osteotomy, non vascularized bone matrix grafting, free vascularized fibular grafts, limited femoral resurfacing, total hip resurfacing, and total hip arthroplasty. PMID- 19294343 TI - RNomics: the new frontier in the post-genomic era. PMID- 19294344 TI - Non-coding RNAs and the acquisition of genomic imprinting in mammals. AB - Genomic imprinting, representing parent-specific expression of alleles at a locus, is mainly evident in flowering plants and placental mammals. Most imprinted genes, including numerous non-coding RNAs, are located in clusters regulated by imprinting control regions (ICRs). The acquisition and evolution of genomic imprinting is among the most fundamental genetic questions. Discoveries about the transition of mammalian imprinted gene domains from their non-imprinted ancestors, especially recent studies undertaken on the most ancient mammalian clades - the marsupials and monotremes from which model species genomes have recently been sequenced, are of high value. By reviewing and analyzing these studies, a close connection between non-coding RNAs and the acquisition of genomic imprinting in mammals is demonstrated. The evidence comes from two observations accompanied with the acquisition of the imprinting: (i) many novel non-coding RNA genes emerged in imprinted regions; (ii) the expressions of some conserved non-coding RNAs have changed dramatically. Furthermore, a systematical analysis of imprinted snoRNA (small nucleolar RNA) genes from 15 vertebrates suggests that the origination of imprinted snoRNAs occurred after the divergence between eutherians and marsupials, followed by a rapid expansion leading to the fixation of major gene families in the eutherian ancestor prior to the radiation of modern placental mammals. Involved in the regulation of imprinted silencing and mediating the chromatins epigenetic modification may be the major roles that non-coding RNAs play during the acquisition of genomic imprinting in mammals. PMID- 19294345 TI - MicroRNA in cell differentiation and development. AB - The regulation of gene expression by microRNAs (miRNAs) is a recently discovered pattern of gene regulation in animals and plants. MiRNAs have been implicated in various aspects of animal development and cell differentiation, such as early embryonic development, neuronal development, muscle development, and lymphocyte development, by the analysis of genetic deletions of individual miRNAs in mammals. These studies show that miRNAs are key regulators in animal development and are potential causes of human diseases. Here we review some recent discoveries about the functions of miRNAs in cell differentiation and development. PMID- 19294346 TI - Are small RNAs a big help to plants? AB - The discovery of RNA interference (RNAi) has augmented our knowledge of gene regulation and presents a fascinating technology that has a great potential for application in genetic analysis, disease therapy, plant protection, and many other areas. In this review, we will focus on the biological functions of RNAi and its application in agriculture with a brief introduction to the history of its discovery and molecular mechanism. PMID- 19294347 TI - New insight into the role of miRNAs in leukemia. AB - Recent studies have shown that microRNAs(miRNAs) play an important role in cell differentiation, growth, and death, including the functional study of miRNAs in tumorigenesis. To date, miRNA expression profiles in many types of cancers have been identified and miRNA expression signatures associated with types and cytogenetics of leukemia have also been reported. Increasing evidence has shown that miRNAs could function as either tumor suppressors or oncogenes in cancers such as leukemia, while other miRNAs might be benefitcial for diagnosis and prognosis, predicted to be newly developed biomarkers. In this review, we summarize the recent progress about miRNAs in leukemia and present a miRNA mediated network involved in differentiation, proliferation and apoptosis predicted to be the roles of miRNAs in the pathogenesis of leukemia. PMID- 19294348 TI - The structure and function of catalytic RNAs. AB - Before the discovery of ribozymes, RNA had been proposed to function as a catalyst, based on the discovery that RNA folded into high-ordered structures as protein did. This hypothesis was confirmed in the 1980s, after the discovery of Tetrahymena group I intron and RNase P ribozyme. There have been about ten ribozymes identified during the past thirty years, as well as the fact that ribosomes function as ribozymes. Advances have been made in understanding the structures and functions of ribozymes, with numerous crystal structures resolved in the past years. Here we review the structure-function relationship of both small and large ribozymes, especially the structural basis of their catalysis. ribozyme, structure, catalysis. PMID- 19294349 TI - tRNAs as regulators in gene expression. AB - Transfer RNAs (tRNAs) hold a central place in protein synthesis by interpreting the genetic information stored in DNA into the amino acid sequence of protein, thus functioning as "adaptor" molecules. In recent years, however, various studies have shown that tRNAs have additional functions beyond participating in protein synthesis. When suffering from certain nutritional stresses, tRNAs change the level of aminoacylation to became uncharged, and these uncharged tRNAs act as effector molecules to regulate global gene expression, so that the stressed organism copes with the adverse environmental stresses. In budding yeast and certain mammalian cells, the retrograde movement of mature tRNAs from cytoplasm to nucleus serves as a mechanism for the surveillance system within the nucleus to continue monitoring the integrity of tRNAs. On the other hand, this retrograde action effectively reduces the global protein synthesis level under conditions of nutritional starvation. Quite recently, various publications have shown that tRNAs are not stable molecules in an absolute sense. Under certain physiological or environmental stresses, they are specifically cleaved into fragments of different lengths in the anticodon loop or anticodon left arm. These cleavages are not a meaningless random degradation phenomenon. Instead, a novel class of signal molecules such as tRNA halves or sitRNAs may be produced, which are closely correlated with the modulation of global gene expression. Investigation of the regulatory functions of tRNAs is a frontier, which seeks to reveal the structural and functional diversity of tRNAs as well as their vital functions during the expression of genetic information. PMID- 19294350 TI - Regulation of mammalian pre-mRNA splicing. AB - In eukaryotes, most protein-coding genes contain introns which are removed by precursor messenger RNA (pre-mRNA) splicing. Alternative splicing is a process by which multiple messenger RNAs (mRNAs) are generated from a single pre-mRNA, resulting in functionally distinct proteins. Recent genome-wide analyses of alternative splicing indicated that in higher eukaryotes alternative splicing is an important mechanism that generates proteomic complexity and regulates gene expression. Mis-regulation of splicing causes a wide range of human diseases. This review describes the current understanding of pre-mRNA splicing and the mechanisms that regulate mammalian pre-mRNA splicing. It also discusses emerging directions in the field of alternative splicing. PMID- 19294351 TI - Diversity and evolution of MicroRNA gene clusters. AB - microRNA (miRNA) gene clusters are a group of miRNA genes clustered within a proximal distance on a chromosome. Although a large number of miRNA clusters have been uncovered in animal and plant genomes, the functional consequences of this arrangement are still poorly understood. Located in a polycistron, the coexpressed miRNA clusters are pivotal in coordinately regulating multiple processes, including embryonic development, cell cycles and cell differentiation. In this review, based on recent progress, we discuss the genomic diversity of miRNA gene clusters, the coordination of expression and function of the clustered miRNAs, and the evolutionarily adaptive processes with gain and loss of the clustering miRNA genes mediated by duplication and transposition events. PMID- 19294352 TI - Protein expression profile in the differentiation of rat bone marrow stromal cells into Schwann cell-like cells. AB - During the last decade, increasing evidence suggested that bone marrow stromal cells (MSCs) have the potential to differentiate into neural lineages. Many studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental conditions. However, no proteomic studies on MSCs differentiated into Schwann cell-like cells have been reported. In this study, we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific conditions. By using two-dimensional gel electrophoresis (2-DE), we compared the protein profiles of MSCs before and after induced differentiation. We obtained 792 protein spots in the protein profile by 2-DE, and found that 74 spots changed significantly before and after the differentiation using PDQuest software, with 43 up-regulated and 31 down-regulated. We analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and by database searching, and found that they could be grouped into various classes, including cytoskeleton and structure proteins, growth factors, metabolic proteins, chaperone proteins, receptor proteins, cell cycle proteins, calcium binding proteins, and other proteins. These proteins also include neural and glial proteins, such as BDNF, CNTF and GFAP. The results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells. PMID- 19294353 TI - Construction of EGFP-tagged rBCG of E.tenella and distribution in chickens. AB - Chicken coccidiosis is a major parasitic disease with substantial economic burden to the poultry industry. Enhanced green fluorescent protein (EGFP) tagged recombinant Bacille Calmette-Guerin (rBCG), as a fusion protein with coccidian rhomboid antigen was constructed to track rBCG in vivo in chickens in this study. Immunization of chickens with one dose of rBCG pMV361-Rho/EGFP induced humoral immune response. The colonization of rBCG in liver, spleen, lung, kidney and caecum was observed by laser confocal microscopy. Real-time quantitative RT-PCR showed a rise expression level of rhomboid protein on the 7th day and a peak on the 14th day and disappearance on the 28th day after immunization. These results have significant implications for the development of rBCG vaccines against avian coccidiosis. PMID- 19294354 TI - Study on Tibetan Chicken embryonic adaptability to chronic hypoxia by revealing differential gene expression in heart tissue. AB - Oxygen concentration is essential for appropriate metabolism. Hypoxia can exert a significant impact on physiological alteration of the cell and organism. Tibetan Chicken (Gallus gallus) is a Chinese indigenous breed inhabiting in Tibetan areas, which is also a chicken breed living at high altitude for the longest time in the world. It has developed an adaptive mechanism to hypoxia, which is demonstrated by that Tibetan Chicken has much higher hatchability than low-land chicken breeds in high-altitude areas of Tibet. In the present study, Tibetan Chicken fertilized full sib eggs were incubated up to Hamburger-Hamilton stage 43 under 13% and 21% oxygen concentration, respectively. Shouguang Chicken and Dwarf Recessive White Chicken were used as control groups. The hearts in all of the 3 chicken breeds under hypoxic and normoxic conditions were isolated and hybridized to GeneChip(R) Chicken Genome Array to study molecular mechanisms underlying the adaptation to high altitude of Tibetan Chicken. As a result, 50 transcripts highly expressed in hypoxia are screened out. Among up-regulated genes, some are involved in the gene ontology (GO) such as cell growth, cell difference, muscle contraction and signal transduction. However, the expression levels of 21 transcripts are lower in hypoxia than those in normoxia. Some down-regulated genes take part in cell communication, ion transport, protein amino acid phosphorylation and signal transduction. Interestingly, gene enrichment analyses of these differential gene expressions are mainly associated with immune system response and ion channel activity in response to stimulus. Moreover, the transcriptional expression profiles analyzed by hierarchical clustering and CPP SOM software in all of the 3 different chicken breeds revealed that Tibetan Chicken is much closely related to Shouguang Chicken rather than Dwarf Recessive White Chicken. In addition, 12 transcripts of Tibetan Chicken breed-specific expressed genes were identified, which seem to result in a more effective and efficient induction of energy demand and signal transduction of transcription and suppression of abnormal development in response to hypoxia. These findings will be beneficial in clarifying the adaptive molecular mechanism of Tibetan Chicken as well as providing new insight into cardiovascular disease at high altitude medicine. PMID- 19294355 TI - Simulation study on the efficiencies of MOET nucleus breeding schemes applying marker assisted selection in dairy cattle. AB - Advantages of breeding schemes using genetic marker information and/or multiple ovulation and embryo transfer (MOET) technology over the traditional approach were extensively evaluated through simulation. Milk yield was the trait of interest and QTL was the genetic marker utilized. Eight dairy cattle breeding scenarios were considered, i.e., traditional progeny testing breeding scheme (denoted as STANPT), GASPT scheme including a pre-selection of young bulls entering progeny testing based on their own QTL information, MOETPT scheme using MOET technology to generate young bulls and a selection of young bulls limited within the full-sib family, GAMOPT scheme adopting both QTL pre-selection and MOET technology, COMBPT scheme using a mixed linear model which considered QTL genotype instead of the BLUP model in GAMOPT, and three non-progeny testing schemes, i.e. the MOET, GAMO and COMB schemes, corresponding to MOETPT, GAMOPT and COMBPT with progeny testing being part of the system. Animals were selected based on their breeding value which was estimated under an animal model framework. Sequential selection over 17 years was performed in the simulations and 30 replicates were designed for each scenario. The influences of using QTL information and MOET technology on favorable QTL allele frequency, true breeding values, polygenetic breeding values and the accumulated genetic superiority were extensively evaluated, for five different populations including active sires, lactating cows, bull dams, bull sires, and young bulls. The results showed that the combined schemes significantly outperformed other approaches wherein accumulated true breeding value progressed. The difference between schemes exclusively using QTL information or MOET technology was not significant. The STANPT scheme was the least efficient among the 8 schemes. The schemes using MOET technology had a higher polygenetic response than others in the 17th year. The increases of frequency of the favorable QTL allele varied more greatly across the 3 male groups than in the lactating cows group. The accumulated genetic superiorities of the GASPT scheme, MOETPT scheme, GAMOPT scheme, COMBPT scheme, MOET scheme, GAMO scheme and COMB scheme over the STANPT scheme were 8.42%, 3.59%, 14.58%, 18.54%, 4.12%, 14.12%, 16.50% in active sires and 2.70%, 5.00%, 11.05%, 12.78%, 7.51%, 17.12%, 25.38% in lactating cows. PMID- 19294358 TI - [Botulinum toxin treatment : therapy success in cases of depression and ongoing pension applications]. AB - INTRODUCTION: A discrepancy is often observed between the presence of botulinum toxin antibodies (Btx-Ab) and successful therapy. We studied whether depression and an ongoing pension application explain this contradiction. METHOD: Sixty-five patients treated with Btx were questioned for subjective therapy failure (TF). The extensor digitorum brevis test (EDB test) indicated the presence of Btx-Ab. The Beck Depression Inventory and an exploration investigated for a depressive disturbance as well as an ongoing application for pension. We correlated the EDB test and the TF with a depressive disturbance and the wish for a pension. RESULTS: Forty-three patients had TF, from which 22 Btx-Ab were positive. A depressive disturbance was found in 25 patients, and an ongoing application for pension in 21. The EDB test and TF did not however correlate TF with a depressive disturbance (P=0.003) and a pension desire (P=0.021). The TF mostly appeared at the beginning of treatment. DISCUSSION: A depressive disturbance and an ongoing application for a pension influence the therapeutic success more strongly initially than the development of Btx-Ab. CONCLUSION: Besides Btx-Ab a depressive disturbance and an ongoing application for pension influences the therapeutic success. PMID- 19294359 TI - [On the 200th birthday of psychiatry]. AB - Two hundred years ago the term "psychiatry" (first named "psychiatery") was introduced by Johann Christian Reil in the city of Halle, Germany. With the word's composition Reil demonstrated that psychic disorders belong to the medical domain, which he justified in a long paper explaining why psychiatry must be one of the three major branches of medicine (the others being surgery and pharmacy). Further he emphasized that psychosomatics and medical psychology are part of the comprehensive new discipline psychiatry. He described psychotherapy as an essential treatment for mental and somatic diseases and having equivalence with pharmacological and surgical methods. Johann Christian Reil launched an antistigma campaign and a crusade towards humanization of psychiatric asylums. Two hundred years later psychiatry is well established as a medical discipline and has become self-confident through its self-determination, in spite of certain setbacks and some objectives that have not yet been achieved. PMID- 19294360 TI - [Orolingual angioedema under systemic thrombolysis with rt-PA: an underestimated side effect]. AB - BACKGROUND: Orolingual angioedema (OA) is an unappreciated complication of acute stroke treatment with recombinant tissue plasminogen activator (rt-PA). It has been described in 2% of patients receiving thrombolysis, and it seems that patients taking angiotensin-converting enzyme inhibitors are especially at risk. Even though the presentation is generally unilateral and limited to lips and tongue, an extension of edema to the oropharynx may lead to life-threatening upper airway obstruction. MATERIAL AND METHODS: In a retrospective analysis of clinical data of 407 patients treated with systemic rt-PA thrombolysis between January 2006 and October 2008 in our department, we describe the occurrence and clinical presentation of OA. RESULTS: Nine of 407 patients (2.2%) showed clinical signs of OA. Typical presentations of OA are illustrated in case reports describing two of these patients and are completed by an overview of the current literature. DISCUSSION: Besides prophylactic inspection of the oral cavity during and after thrombolysis, therapeutic options in case of OA include early intravenous antihistaminergic therapy and protective intubation. PMID- 19294361 TI - [Medical treatment of epilepsy: hidden dimensions]. AB - Treatment with antiepileptic drugs (AED) increases the chance of seizure control in patients with epilepsy. This control is best achieved with AED monotherapy in the early stages of the disease. In up to 28% of the remaining patients, complete seizure control is obtained with AED combination therapy. In our own retrospective studies, levetiracetam proved to be the most effective drug in add on treatment of focal epilepsies. Seizure control for at least 6 months was seen in 22.8% of patients, and reduction of seizure frequency by more than 50% was achieved in 58.8%. One must however consider that up to 50% of all seizures are not recognized by the patients. This is especially the case in complex partial seizures and in seizures during sleep. Seizure control (as recognized by the patient) allows better social reintegration, e.g. by obtaining a driving license. In addition to AED therapy other epilepsy-related symptoms such as depression need diagnosis and treatment. Of all patients with epilepsy, 30-50% suffer from depression, but about 50% of these are not willing to start antidepressant drug therapy. PMID- 19294362 TI - Optimal dosing of renin-angiotensin-aldosterone system blockers for renal protection: a solved issue? PMID- 19294363 TI - Islet beta cell failure in the 60% pancreatectomised obese hyperlipidaemic Zucker fatty rat: severe dysfunction with altered glycerolipid metabolism without steatosis or a falling beta cell mass. AB - AIMS/HYPOTHESIS: The Zucker fatty (ZF) rat subjected to 60% pancreatectomy (Px) develops moderate diabetes by 3 weeks. We determined whether a progressive fall in beta cell mass and/or beta cell dysfunction contribute to beta cell failure in this type 2 diabetes model. METHODS: Partial (60%) or sham Px was performed in ZF and Zucker lean (ZL) rats. At 3 weeks post-surgery, beta cell mass and proliferation, proinsulin biosynthesis, pancreatic insulin content, insulin secretion, and islet glucose and lipid metabolism were measured. RESULTS: ZL-Px rats maintained normal glycaemia and glucose-stimulated insulin secretion (GSIS) despite incomplete recovery of beta cell mass possibly due to compensatory enhanced islet glucose metabolism and lipolysis. ZF-Px rats developed moderate hyperglycaemia (14 mmol/l), hypertriacylglycerolaemia and relative hypoinsulinaemia. Despite beta cell mass recovery and normal arginine-induced insulin secretion, GSIS and pancreatic insulin content were profoundly lowered in ZF-Px rats. Proinsulin biosynthesis was not reduced. Compensatory increases in islet glucose metabolism above those observed in ZF-Sham rats were not seen in ZF Px rats. Triacylglycerol content was not increased in ZF-Px islets, possibly due to lipodetoxification by enhanced lipolysis and fatty acid oxidation. Fatty acid accumulation into monoacylglycerol and diacylglycerol was increased in ZF-Px islets together with a 4.5-fold elevation in stearoyl-CoA desaturase mRNA expression. CONCLUSIONS/INTERPRETATION: Falling beta cell mass, reduced proinsulin biosynthesis and islet steatosis are not implicated in early beta cell failure and glucolipotoxicity in ZF-Px rats. Rather, severe beta cell dysfunction with a specific reduction in GSIS and marked depletion of beta cell insulin stores with altered lipid partitioning underlie beta cell failure in this animal model of type 2 diabetes. PMID- 19294364 TI - Humidification during oxygen therapy and non-invasive ventilation: do we need some and how much? PMID- 19294365 TI - Discomfort associated with underhumidified high-flow oxygen therapy in critically ill patients. AB - OBJECTIVE: To measure (1) the discomfort in non-intubated patients under high flow oxygen therapy (HFOT) humidified with bubble (BH) or heated humidifiers (HH), and (2) the hygrometric properties of oxygen with a BH and an HH. DESIGN AND SETTING: This was a randomized cross-over study in critically ill patients during a 3-day period. The humidification device used at days 1 and 3 was changed for the other at day 2. (2) It was also an experimental bench study using the psychrometric method with five randomized flows (3, 6, 9, 12 and 15 l/min) and different humidification techniques. METHODS: Discomfort, particularly dryness of the mouth and throat, was measured for two humidification conditions (BH and HH) using a 0-10 numerical rating scale (NRS) by patients requiring HFOT with a face mask at a flow >/=5 l/min, in a double-blinded condition. RESULTS: (1) In this clinical study, 30 patients treated by HFOT at a median flow of 7.8 l/min (5.1 10.9) were included. The global incidence of moderate (NRS = 4-6) and severe discomfort (NRS = 7-10) was 25 and 29%, respectively. The median intensities of both mouth and throat dryness were significantly lower with the HH than with the BH [7.8 (5.0-9.4) vs. 5.0 (3.1-7.0), P = 0.001 and 5.8 (2.3-8.5) vs. 4.3 (2.0 5.0), P = 0.005, respectively]. (2) In the bench study, the mean absolute humidity measured at an ambient temperature of 26 degrees C with the HH was two times greater than with the BH (30 +/- 1 vs. 16 +/- 2 mg/l, P < 0.05) regardless of the flow rate. CONCLUSIONS: Compared to bubble humidifiers, the use of a heated-humidifier in patients with high-flow oxygen therapy is associated with a decrease of dryness symptoms mediated by increased humidity delivered to the patient. PMID- 19294366 TI - Water content of delivered gases during non-invasive ventilation in healthy subjects. AB - INTRODUCTION: No clear recommendation exists concerning humidification during non invasive ventilation (NIV) and high flow CPAP, and few hygrometric data are available. METHODS: We measured hygrometry during NIV delivered to healthy subjects with different humidification strategies: heated humidifier (HH), heat and moisture exchanger, (HME) or no humidification (NoH). For each strategy, a turbine and an ICU ventilator were used with different FiO(2) settings, with and without leaks. During CPAP, two different HH and NoH were tested. Inspired gases hygrometry was measured, and comfort was assessed. On a bench, we also assessed the impact of ambient air temperature, ventilator temperature and minute ventilation on HH performances (with NIV settings). RESULTS: During NIV, with NoH, gas humidity was very low when an ICU ventilator was used (5 mgH(2)O/l), but equivalent to ambient air hygrometry with a turbine ventilator at minimal FiO(2) (13 mgH(2)O/l). HME and HH had comparable performances (25-30 mgH(2)O/l), but HME's effectiveness was reduced with leaks (15 mgH(2)O/l). HH performances were reduced by elevated ambient air and ventilator output temperatures. During CPAP, dry gases (5 mgH(2)O/l) were less tolerated than humidified gases. Gases humidified at 15 or 30 mgH(2)O/l were equally tolerated. CONCLUSION: This study provides data on the level of humidity delivered with different humidification strategies during NIV and CPAP. HH and HME provide gas with the highest water content. Comfort data suggest that levels above 15 mgH(2)O/l are well tolerated. In favorable conditions, HH and HMEs are capable of providing such values, even in the presence of leaks. PMID- 19294367 TI - Certified reference material for quantification of polycyclic aromatic hydrocarbons in sediment from the National Metrology Institute of Japan. AB - The National Metrology Institute of Japan has issued a certified reference material (CRM) of freshwater lake sediment for polycyclic aromatic hydrocarbon (PAHs) analyses. The certification used three extraction techniques: pressurized liquid extraction (PLE) with toluene, PLE with dichloromethane/ethyl acetate (1:1 by volume), and alkaline extraction (1 M KOH in methanol) in combination with microwave-assisted extraction. Both gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/dopant-assisted atmospheric pressure photoionization/MS (LC/DA-APPI/MS) analyses were used. Certified values are provided for 18 PAHs at 1-25 microg kg(-1) except for perylene (2.08 x 10(3) microg kg(-1)), and information values are provided for two. Since the values of PAHs in the CRM are much lower than those in other CRMs and are comparable to those found at sites with little human influence, the CRM is suitable for PAH monitoring in sediment and soil samples. PMID- 19294368 TI - Complex surface analytical investigations on hydrogen absorption and desorption processes of a TiMn2-based alloy. AB - Metal hydrides are one of the most promising technologies in the field of hydrogen storage due to their high volumetric storage density. Important reaction steps take place at the very surface of the solid during hydrogen absorption. Since these reaction steps are drastically influenced by the properties and potential contamination of the solid, it is very important to understand the characteristics of the surface, and a variety of analytical methods are required to achieve this. In this work, a TiMn(2)-type metal hydride alloy is investigated by means of high-pressure activation measurements, X-ray photoelectron spectroscopy (XPS), secondary neutral mass spectrometry (SNMS) and thermal desorption mass spectrometry (TDMS). In particular, TDMS is an analytical tool that, in contrast to SIMS or SNMS, allows the hydrogen content in a metal to be quantified. Furthermore, it allows the activation energy for desorption to be determined from TDMS profiles; the method used to achieve this is presented here in detail. In the results section, it is shown that the oxide layer formed during manufacture and long-term storage prevents any hydrogen from being absorbed, and so an activation process is required. XPS measurements show the oxide states of the main alloy elements, and a layer 18 nm thick is determined via SNMS. Furthermore, defined oxide layers are produced and characterized in UHV using XPS. The influence of these thin oxide layers on the hydrogen sorption process is examined using TDMS. Finally, the activation energy of desorption is determined for the investigated alloy using the method presented here, and values of 46 kJ/mol for hydrogen sorbed in UHV and 103 kJ/mol for hydrogen originating from the manufacturing process are obtained. PMID- 19294369 TI - Learning the visuomotor transformation of virtual and real sliding levers: simple approximations of complex transformations. AB - Learning to operate a complex tool such as a sliding lever can be conceived as learning both a kinematic and a dynamic transformation. We investigated whether the presence of the dynamic transformation has an inhibitory or a facilitative effect on learning to control a sliding lever. Furthermore, we examined the characteristics of the internal model of the kinematic transformation of the lever in visual open-loop trials. In the experiment, one group of participants practiced with only the kinematic transformation of the lever (virtual lever), the other group practiced with both the kinematic and the dynamic transformation (physical lever). Visual feedback was continuously present during practice. Results showed only marginal differences between both groups in open-loop tests. This finding is likely to be related to the fact that in both groups a simplified approximation of the kinematic transformation was acquired, in particular a symmetry approximation. With such an approximation the target for the hand movement is derived from the visual target for the tip of the lever as the position which is symmetric around a sagittal axis. PMID- 19294370 TI - The role of TRPV1 receptors in pain evoked by noxious thermal and chemical stimuli. AB - Transient receptor potential receptors (TRP) on primary afferent neurons respond to noxious and/or thermal stimuli. TRPV1 receptors can be activated by noxious heat, acid, capsaicin and resiniferatoxin, leading to burning pain or itch mediated by discharges in C polymodal and Adelta mechano-heat nociceptors and in central neurons, including spinothalamic tract (STT) cells. Central nociceptive transmission involves both non-NMDA and NMDA receptors, and inhibitory interneurons as well as projection neurons contribute to the neural interactions. Behavioral consequences of intradermal injection of capsaicin include pain, as well as primary and secondary hyperalgesia and allodynia. Primary hyperalgesia depends on sensitization of peripheral nociceptors, whereas, secondary hyperalgesia and allodynia result from sensitization of central nociceptive neurons, such as STT cells. Central sensitization is associated with enhanced responses to excitatory amino acids and decreased responses to inhibitory amino acids. The mechanism of the increase in responses to excitatory amino acids includes phosphorylation of NR1 subunits of NMDA receptors and GluR1 subunits of AMPA receptors. Central sensitization depends on activation of several protein kinases and other enzymes, such as nitric oxide synthase. This process is regulated by protein phosphatases. Central sensitization can be regarded as a spinal cord form of long-term potentiation. PMID- 19294371 TI - Regional surveillance of emergency-department visits for outpatient adverse drug events. AB - AIMS: To determine the (1) incidence of adverse drug events (ADEs) in 10 emergency department (EDs) of general hospitals in the Regione Campania (southern Italy), (2) rate of ADE-related hospital admissions, (3) drug classes most frequently involved, and (4) the types of ADEs and their frequency. METHODS: We performed a cohort study of all patients attending the EDs. This study was carried out in two observational periods of 10 days each in 10 EDs. Demographic, clinical, and pharmacological data about all patients admitted to EDs were collected by trained and qualified monitors. Records related to ADEs were analyzed and validated by a specific scientific committee. RESULTS: Of 7,861 ED visits, 96 were ADE-related. The incidence of hospitalization was higher in patients who had taken medication than in patients with a negative drug history (24.9 vs. 16.4%). ADEs were significantly more frequent in women. Patients aged between 60 and 69 years and between 30 and 39 years were significantly more likely to experience an ADE. Serious ADEs were identified in 20 ED visits (20.8% of total sample). Antibiotics, NSAIDs, and agents acting on the renin-angiotensin system were the drugs most often involved in ADEs. In multivariate analyses, the adjusted odds ratio was 3.4 (95% CI: 1.07-2.84) for patients taking NSAIDs, 4.78 (95% CI: 2.26-10.12) for those taking beta(2)-adrenergic-receptor agonists, and 6.20 (95%CI: 2.74-14.06) for those taking beta-lactam antibiotics. CONCLUSION: This study shows that ADEs are an important problem in industrialized countries. Moreover, it shows that ADEs affect hospital admission rates and reinforces the importance of drug-induced disease as a public health problem. PMID- 19294372 TI - MRI appearance of Rasmussen encephalitis. PMID- 19294373 TI - Histology of the dural membrane supports the theoretical considerations of its role in the pathophysiology of subdural collections in nontraumatic circumstances. PMID- 19294374 TI - HLA-A allele associations with viral MER9-LTR nucleotide sequences at two distinct loci within the MHC alpha block. AB - The study of the association of the Human Leukocyte Antigen (HLA) alleles and polymorphic retrotransposons such as Alu, HERV, and LTR at various loci within the Major Histocompatibility Complex allows for a better identification and stratification of disease associations and the origins of HLA haplotypes in different populations. This paper provides sequence and association data on two structurally polymorphic MER9-LTR retrotransposons that are located 54 kb apart and in close proximity to the multiallelic HLA-A gene involved in the regulation of the human immune system. Direct DNA sequencing and analysis of the PCR products identified DNA nucleotide variations between the MER9-LTR sequences at the two loci and their associations with HLA-A alleles as potential haplotype and evolutionary markers. All MER9-LTR sequences were haplotypic when associated with common HLA-A alleles. The number of SNP loci was 2.5 times greater for the solo LTR at the AK locus, which is located closer to the HLA-A gene than the solo or 3' LTR at the HG locus. Our study shows that the nucleotide variations of the MER9-LTR DNA sequences are additional informative markers in fine mapping HLA-A genomic haplotypes for future population, evolutionary, and disease studies. PMID- 19294375 TI - Synthesis and characterization of hydroquinone fructoside using Leuconostoc mesenteroides levansucrase. AB - Hydroquinone (HQ) functions as a skin-whitening agent, but it has the potential to cause dermatitis. We synthesized a HQ fructoside (HQ-Fru) as a potential skin whitening agent by reacting levansucrase from Leuconostoc mesenteroides with HQ as an acceptor and sucrose as a fructofuranose donor. The product was purified using 1-butanol partition and silica-gel column chromatography. The structure of the purified HQ-Fru was determined by (1)H and (13)C nuclear magnetic resonance, and the molecular ion of the product was observed at m/z 295 (C12 H16 O7 Na)(+). The HQ-Fru was identified as 4-hydroxyphenyl-beta-D: -fructofuranoside. The optimum condition for HQ-Fru synthesis was determined using a response surface method (RSM), and the final optimum condition was 350 mM HQ, 115 mM sucrose, and 0.70 U/ml levansucrase, and the final HQ-Fru produced was 1.09 g/l. HQ-Fru showed anti-oxidation activities and inhibition against tyrosinase. The median inhibition concentration (IC(50)) of 1,1-diphenyl-2-picrylhydrazyl scavenging activity was 5.83 mM, showing higher antioxidant activity compared to beta arbutin (IC(50) = 6.04 mM). The K ( i ) value of HQ-Fru (1.53 mM) against tyrosinase was smaller than that of beta-arbutin (K ( i ) = 2.8 mM), indicating that it was 1.8-times better as an inhibitor. The inhibition of lipid peroxidation by HQ-Fru was 105.3% that of HQ (100%) and 118.9 times higher than that of beta-arbutin (0.89% of HQ). PMID- 19294376 TI - Giant cell tumour of the triquetrum. AB - We present details of a case of giant cell tumour of bone (GCTOB) involving the triquetrum. GCTOB arising within the carpus is exceedingly rare and, to our knowledge, this is only the second case of monostotic GCTOB of the triquetrum that has been reported. PMID- 19294377 TI - Detection of partial-thickness supraspinatus tendon tears: is a single direct MR arthrography series in ABER position as accurate as conventional MR arthrography? AB - PURPOSE: The purpose of this study was to retrospectively evaluate sensitivity and specificity of a single magnetic resonance (MR) arthrography series in abduction external rotation (ABER) position compared with conventional MR arthrography for detection of supraspinatus tendon tears, with arthroscopy as gold standard, and to assess interobserver variability. MATERIALS AND METHODS: Institutional review board approval was obtained; informed consent was waived. MR arthrograms of 250 patients (170 men and 80 women; mean age, 36 years) were retrospectively and independently evaluated by three observers. Oblique coronal T1-weighted fat-suppressed images, proton density, and T2-weighted images and axial T1-weighted images and oblique sagittal T1-weighted fat-suppressed images were analyzed to detect supraspinatus tendon tears. Separately, a single T1 weighted fat-suppressed oblique axial series in ABER position was evaluated. Both protocols were scored randomly without knowledge of patients' clinical history and arthroscopy results. Tears were subclassified, based on articular surface integrity and extension (Lee classification). Interobserver agreement was assessed by kappa statistics for all patients. Ninety-two of 250 patients underwent arthroscopy; sensitivity and specificity of ABER and conventional MR arthrography were calculated and compared using paired McNemar test. RESULTS: Weighted kappa values of ABER and conventional MR arthrography were 0.48-0.65 and 0.60-0.67, respectively. According to arthroscopy, 69 of 92 patients had an intact cuff, and 23 patients had a cuff tear (16 partial thickness and seven full thickness). There were no statistically significant differences between ABER and conventional MR arthrography regarding sensitivity (48-61% and 52-70%, respectively) and specificity (80-94% and 91-95%). CONCLUSION: Sensitivity and specificity of a single T1-weighted series in ABER position and conventional MR arthrography are comparable for assessment of rotator cuff tears. PMID- 19294378 TI - Fast MR arthrography using VIBE sequences to evaluate the rotator cuff. AB - PURPOSE: The purpose of this paper was to evaluate if short volumetric interpolated breath-hold examination (VIBE) sequences can be used as a substitute for T1-weighted with fat saturation (T1-FS) sequences when performing magnetic resonance (MR) arthrography to diagnose rotator cuff tears. MATERIALS AND METHODS: Eighty-two patients underwent direct MR arthrography of the shoulder joint using VIBE (acquisition time of 13 s) and T1-FS (acquisition time of 5 min) sequences in the axial and paracoronal plane on a 1.0-T MR unit. Two radiologists scored rotator cuff tendons on VIBE and T1-FS images separately as normal, small/large partial thickness and full thickness tears with or without geyser sign. T1-FS sequences were considered the gold standard. Surgical correlation was available in a small sample. RESULTS: Sensitivity, specificity, and positive and negative predictive values of VIBE were greater than 92% for large articular sided partial thickness and full thickness tears. For detecting fraying and articular-sided small partial thickness tears, these parameters were 55%, 94%, 94%, and 57%, respectively. The simple kappa value was 0.76, and the weighted kappa value was 0.86 for agreement between T1-FS and VIBE scores. All large partial and full thickness tears at surgery were correctly diagnosed using VIBE or T1-FS MR images. CONCLUSION: Fast MR arthrography of the shoulder joint using VIBE sequences showed good concordance with the classically used T1-FS sequences for the appearance of the rotator cuff, in particular for large articular-sided partial thickness tears and for full thickness tears. Due to its very short acquisition time, VIBE may be especially useful when performing MR arthrography in claustrophobic patients or patients with a painful shoulder. PMID- 19294379 TI - Assessment of cartilage-dedicated sequences at ultra-high-field MRI: comparison of imaging performance and diagnostic confidence between 3.0 and 7.0 T with respect to osteoarthritis-induced changes at the knee joint. AB - OBJECTIVE: The objectives of the study were to optimize three cartilage-dedicated sequences for in vivo knee imaging at 7.0 T ultra-high-field (UHF) magnetic resonance imaging (MRI) and to compare imaging performance and diagnostic confidence concerning osteoarthritis (OA)-induced changes at 7.0 and 3.0 T MRI. MATERIALS AND METHODS: Optimized MRI sequences for cartilage imaging at 3.0 T were tailored for 7.0 T: an intermediate-weighted fast spin-echo (IM-w FSE), a fast imaging employing steady-state acquisition (FIESTA) and a T1-weighted 3D high-spatial-resolution volumetric fat-suppressed spoiled gradient-echo (SPGR) sequence. Three healthy subjects and seven patients with mild OA were examined. Signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), diagnostic confidence in assessing cartilage abnormalities, and image quality were determined. Abnormalities were assessed with the whole organ magnetic resonance imaging score (WORMS). Focal cartilage lesions and bone marrow edema pattern (BMEP) were also quantified. RESULTS: At 7.0 T, SNR was increased (p < 0.05) for all sequences. For the IM-w FSE sequence, limitations with the specific absorption rate (SAR) required modifications of the scan parameters yielding an incomplete coverage of the knee joint, extensive artifacts, and a less effective fat saturation. CNR and image quality were increased (p < 0.05) for SPGR and FIESTA and decreased for IM w FSE. Diagnostic confidence for cartilage lesions was highest (p < 0.05) for FIESTA at 7.0 T. Evaluation of BMEP was decreased (p < 0.05) at 7.0 T due to limited performance of IM-w FSE. CONCLUSION: Gradient echo-based pulse sequences like SPGR and FIESTA are well suited for imaging at UHF which may improve early detection of cartilage lesions. However, UHF IM-w FSE sequences are less feasible for clinical use. PMID- 19294381 TI - Phase I clinical study of anti-apoptosis protein survivin-derived peptide vaccination for patients with advanced or recurrent urothelial cancer. AB - Survivin, a member of the inhibitor of apoptosis protein family, is expressed in many malignant tumors including urothelial cancer but is hardly detectable in normal, differentiated adult tissues. Previously we reported CD8-positive cytotoxic T-lymphocytes (CTLs) were successfully induced by stimulation with survivin-2B80-88 peptide in vitro. We started a phase I clinical study of survivin-2B80-88 peptide vaccination for advanced urothelial cancer patients to assess the safety and efficacy of this vaccination. Nine patients were received vaccination and were evaluated for immunological evaluation, adverse events, and clinical responses. A total of 46 vaccinations were carried out. There was no severe adverse event. HLA-A24/survivin-2B80-88 peptide tetramer analysis revealed a significant increase in the peptide-specific CTL frequency after the vaccination in five patients. Slight reduction of the tumor volume was observed in one patient. Survivin-2B80-88 peptide-based vaccination is safe and should be further considered for potential immune and clinical efficacy in urothelial cancer patients. PMID- 19294382 TI - Onconeural antibodies in sera from patients with various types of tumours. AB - PURPOSE: We assessed the frequency and levels of onconeural antibodies in 974 patients with various types of tumours, but without apparent paraneoplastic neurological syndromes (PNS). PATIENTS AND METHODS: We included patients with the following tumours: 200 small-cell lung cancer (SCLC) patients, 253 breast cancer patients, 182 ovarian cancer patients, 266 uterine cancer patients and 73 thymoma patients, as well as 52 patients with PNS and cancer and 300 healthy blood donors. Sera were screened for amphiphysin, CRMP5, Hu, Ma2, Ri and Yo antibodies using a multi-well immunoprecipitation technique. RESULTS: The most frequently detected antibodies were Hu followed by CRMP5. Ma2, Yo, amphiphysin and Ri antibodies were less common, but each was found at similar frequencies. Onconeural antibodies were present at similar levels in sera from the PNS control group and from cancer patients. Hu antibodies were rare in cancers other than SCLC. CRMP5 was the only antibody found in patients with thymoma and this antibody was more common among patients with thymoma than in other tumour patients. With one exception, coexisting antibodies were only found in patients with SCLC. The presence of onconeural antibodies in SCLC patients was not associated with prolonged survival. CONCLUSION: Onconeural antibodies are associated with various types of tumours suggesting that all antibodies should be included in the serological screening for possible PNS. The levels of onconeural antibody are not sufficiently sensitive to discriminate between cancer patients with PNS and those without. PMID- 19294383 TI - Oral administration of poly-gamma-glutamate induces TLR4- and dendritic cell dependent antitumor effect. AB - Previously, we reported that the oral administration of high molecular mass poly gamma-glutamate (gamma-PGA) induced antitumor immunity but the mechanism underlying this antitumor activity was not understood. In the present study, we found that application of high molecular mass gamma-PGA induced secretion of tumor necrosis factor (TNF)-alpha from the bone-marrow-derived macrophages of wild type (C57BL/6 and C3H/HeN) and Toll-like receptor 2 knockout (TLR2(-/-)) mice, but not those of myeloid differentiation factor 88 knockout (MyD88(-/-)) and TLR4-defective mice (C3H/HeJ). Production of interferon (IFN)-gamma-inducible protein 10 (IP-10) in response to treatment with gamma-PGA was almost abolished in C3H/HeJ mice. In contrast to LPS, gamma-PGA induced productions of TNF-alpha and IP-10 could not be blocked by polymyxin B. Furthermore, gamma-PGA-induced interleukin-12 production was also impaired in immature dendritic cells (iDCs) from MyD88(-/-) and C3H/HeJ mice. Downregulation of MyD88 and TLR4 expression using small interfering RNA (siRNA) significantly inhibited gamma-PGA-induced TNF alpha secretion from the RAW264.7 cells. Gamma-PGA-mediated intracellular signaling was markedly inhibited in C3H/HeJ cells. The antitumor effect of gamma PGA was completely abrogated in C3H/HeJ mice compared with control mice (C3H/HeN) but significant antitumor effect was generated by the intratumoral administration of C3H/HeN mice-derived iDCs followed by 2,000 kDa gamma-PGA in C3H/HeJ. These findings strongly suggest that the antitumor activity of gamma-PGA is mediated by TLR4. PMID- 19294384 TI - Treatment of non-Hodgkin's lymphoma xenografts with the HB22.7 anti-CD22 monoclonal antibody and phosphatase inhibitors improves efficacy. AB - PURPOSE: To examine the role of phosphatase inhibition on anti-CD22, HB22.7 mediated lymphomacidal effects. EXPERIMENTAL DESIGN: CD22 is a cell-surface molecule expressed on most B cell lymphomas (NHL). HB22.7 is an anti-CD22 monoclonal antibody that binds a unique CD22-epitope, blocks ligand binding, initiates signaling, and has demonstrated lymphomacidal activity. The SHP-1 tyrosine phosphatase is associated with the cytoplasmic domain of CD22. Sodium orthovanadate (NaV) is a phosphatase inhibitor. The SHP-1-CD22 interaction presents an opportunity to manipulate CD22-mediated signaling effects. In vitro cell culture assays and in vivo human NHL xenograft studies were used to assess the effects of phosphatase inhibition. RESULTS: NaV caused dose dependent killing of NHL cells in vitro; when HB22.7 was given with NaV, antibody-mediated cell death was augmented. Flow cytometry showed that NaV-pretreatment resulted in less CD22 internalization after ligation with HB22.7 than did control cells. Studies in mice bearing Raji NHL xenografts showed that the combination of NaV and HB22.7 shrank NHL tumors more rapidly, had a higher complete response rate (80%), and produced the best survival compared to controls; no toxicity was detected. Studies using Raji cells stably transfected with SHP-1DN confirmed that these observations were due to SHP-1 inhibition. CONCLUSION: The relatively specific association of SHP-1 with CD22 suggests that CD22-specific signal augmentation by phosphatase inhibitors can improve the clinical outcome of anti-CD22 based immunotherapy. PMID- 19294385 TI - Risk factors for leukemia in Thailand. AB - A case-control study of adult-onset leukemia was conducted in Bangkok, Thailand to explore the contribution of cellular telephone use and other factors to the etiology of the disease; 180 cases (87 acute myeloblastic leukemia, 40 acute lymphoblastic leukemia, 44 chronic myelogenous leukemia, eight chronic lymphocytic leukemia, one unclassified acute leukemia) were compared with 756 age and sex-matched hospital controls. Data were obtained by interview; odds ratios (ORs) were estimated by unconditional logistic regression. There was no clear association with cellular telephone phone use, but durations were relatively short (median 24-26 months), and there was a suggestion that risk may be increased for those with certain usage practices (ORs, 1.8-3.0 with lower confidence intervals >1.0) and those who used GSM service (OR, 2.1; 95% confidence interval, 1.1-4.0). Myeloid leukemia (acute and chronic combined) was associated with benzene (OR, 3.9; 95% confidence interval, 1.3-11), a nonspecific group of other solvents (2.3; 1.1-4.9), occupational pesticides that were mostly unspecified (3.8; 2.1-7.1), and working with or near powerlines (4.3; 1.3-15). No associations were found for diagnostic X-rays, cigarette smoking, or other occupational exposures. PMID- 19294386 TI - Do anthocyanins and anthocyanidins, cancer chemopreventive pigments in the diet, merit development as potential drugs? AB - Anthocyanins, plant pigments in fruits and berries, have been shown to delay cancer development in rodent models of carcinogenesis, especially those of the colorectal tract. Anthocyanins and anthocyanidins, their aglycons, especially cyanidin and delphinidin, have been subjected to extensive mechanistic studies. In cells in vitro, both glycosides and aglycons engage an array of anti-oncogenic mechanisms including anti-proliferation, induction of apoptosis and inhibition of activities of oncogenic transcription factors and protein tyrosine kinases. Anthocyanins and anthocyanidins exist as four isomers, interconversion between which depends on pH, temperature and access to light. Anthocyanidins are much more prone to avid chemical decomposition than the glycosides, and they only survive for minutes in the biophase. These pharmaceutical issues are very important determinants of the suitability of these flavonoids for potential development as cancer chemopreventive drugs, and they have hitherto not received adequate attention. In the light of their robust cancer chemopreventive efficacy in experimental models and their superior stability as compared to that of the aglycons, the anthocyanins seem much more suitable for further drug development than their anthocyanidin counterparts. PMID- 19294387 TI - A randomized, phase II, dose-finding study of the pan-ErbB receptor tyrosine kinase inhibitor CI-1033 in patients with pretreated metastatic breast cancer. AB - PURPOSE: To evaluate the efficacy and safety of the pan-ErbB receptor tyrosine kinase inhibitor CI-1033 in metastatic breast cancer (MBC). EXPERIMENTAL DESIGN: Patients with measurable, progressive, or recurrent MBC whose primary tumor expressed > or =1 ErbB receptor were randomized to the following CI-1033 regimens: 50 mg (arm A) or 150 mg (arm B) daily without rest period, or 450 mg/day x 14 days every 21 days (arm C). The primary endpoint was 1-year progression-free survival (PFS). RESULTS: Overall, 194 patients were treated. One year PFS estimates were 3.8, 2.0, and 4.6%; median PFS was 61, 56, and 58 days; and investigator-assessed overall response rates were 1.5, 1.5, and 7.3%, in arms A, B, and C, respectively. Response duration was 110-419 days. In arm C, response (18.8 vs. 2.6%) and 1-year overall survival rates (86.7 vs. 47.5%) were greater in patients with HER2-positive versus HER2-negative tumors. The incidence of grade 3/4 adverse events (AEs) was dose-dependent, affecting 10.3, 48.6, and 80.4% of patients in arms A, B and C, respectively. The most common grade 3/4, treatment-related AEs were diarrhea, asthenia, and stomatitis. Arm C enrollment was prematurely discontinued due to a high frequency of grade 3/4 AEs. CONCLUSION: Single-agent CI-1033 did not show clinically meaningful activity in heavily pretreated patients with MBC expressing > or =1 ErbB receptor. Antitumor activity was observed in arm C patients with HER2-positive tumors. However, only the 50 mg dose was well tolerated, and the highest dose reached unacceptable levels of toxicity. PMID- 19294388 TI - Should previous mammograms be digitised in the transition to digital mammography? AB - Breast screening specificity is improved if previous mammograms are available, which presents a challenge when converting to digital mammography. Two display options were investigated: mounting previous film mammograms on a multiviewer adjacent to the workstation, or digitising them for soft copy display. Eight qualified screen readers were videotaped undertaking routine screen reading for two 45-min sessions in each scenario. Analysis of gross eye and head movements showed that when digitised, previous mammograms were examined a greater number of times per case (p = 0.03), due to a combination of being used in 19% more cases (p = 0.04) and where used, looked at a greater number of times (28% increase, p = 0.04). Digitising previous mammograms reduced both the average time taken per case by 18% (p = 0.04) and the participants' perceptions of workload (p < 0.05). Digitising previous analogue mammograms may be advantageous, in particular in increasing their level of use. PMID- 19294389 TI - Triple therapy in refractory detrusor overactivity: a preliminary study. AB - PURPOSE: To evaluate in a prospective study the impact of the "three-drug therapy" (antimuscarinic, alpha-blocker and tricyclic antidepressants) on the treatment of refractory detrusor overactivity (DO). METHODS: Data from 27 consented patients with refractory DO were available for study. They were asked to complete a daily urinary chart and underwent urodynamic evaluation (UD) before and 60 days after treatment. Response to treatment was considered the presence of one or less involuntary detrusor contractions (IDC) on post-treatment UD. Statistical analysis was performed with Fisher and Mann-Whitney tests, besides Spearman's correlation. P values <0.05 were considered significant. RESULTS: The mean follow-up was 15 months. The comparison of the daily urinary chart before and after treatment showed significant increase on bladder capacity and decreases on urgency, urge-incontinence and frequency. Objective data from UD showed that the mean maximum bladder capacity (MBC) ranged from 200 to 300 mL (P < 0.001) with treatment. The same trend was observed with the other UD variables. When compared to baseline, the questionnaire OAB-v8 showed significant improvement (P < 0.01). Main side effects comprised dry mouth and constipation (40%), with average scores of 5.16 and 3.08, respectively (visual scale from 0 to 10). CONCLUSIONS: Triple therapy may be an effective, easily employed and well tolerated option to refractory DO treatment. More studies are necessary to achieve more consistent data on the matter. PMID- 19294390 TI - Cuttlefish use visual cues to control three-dimensional skin papillae for camouflage. AB - Cephalopods (octopus, squid and cuttlefish) are known for their camouflage. Cuttlefish Sepia officinalis use chromatophores and light reflectors for color change, and papillae to change three-dimensional physical skin texture. Papillae vary in size, shape and coloration; nine distinct sets of papillae are described here. The objective was to determine whether cuttlefish use visual or tactile cues to control papillae expression. Cuttlefish were placed on natural substrates to evoke the three major camouflage body patterns: Uniform/Stipple, Mottle and Disruptive. Three versions of each substrate were presented: the actual substrate, the actual substrate covered with glass (removes tactile information) and a laminated photograph of the substrate (removes tactile and three dimensional information because depth-of-field information is unavailable). No differences in Small dorsal papillae or Major lateral mantle papillae expression were observed among the three versions of each substrate. Thus, visual (not tactile) cues drive the expression of papillae in S. officinalis. Two sets of papillae (Major lateral mantle papillae and Major lateral eye papillae) showed irregular responses; their control requires future investigation. Finally, more Small dorsal papillae were shown in Uniform/Stipple and Mottle patterns than in Disruptive patterns, which may provide clues regarding the visual mechanisms of background matching versus disruptive coloration. PMID- 19294391 TI - Intradural arteriosynangiosis in pediatric moyamoya disease: modified technique of encephalo-duro-arterio-synangiosis with reduced operative damage to already growing revascularization. AB - OBJECTIVE: Various operative procedures have been described for the treatment of pediatric moyamoya disease. However, the majority of invasive or radical procedures proposed have focused primarily on revascularization and few have discussed to maximizing preservation of the already growing neovascular network. METHODS: This present procedure describes the supratemporal artery is anastomosed to the inner layer of the dura mater and surrounded by the outer layer as a sandwich with a blunt procedure of dural layer separation. RESULTS: This technique efficiently established an anastomotic connection between the supratemporal artery and the cortical brain, and the dura mater postoperatively with maximally preserving the existing vascular network. CONCLUSION: IDAS, the modified surgical technique of EDAS, shall be benefit in preserving the already developed collateral circulation, and underlying structure over the brain cortex as much as possible, which is important particularly in pediatric moyamoya patients. PMID- 19294392 TI - Factors predicting the postoperative outcome of lower gastrointestinal hemorrhage. AB - PURPOSE: To examine the treatment outcome for patients with acute bleeding from the lower gastrointestinal tract requiring transfusion and acute surgical care as a function of various risk factors MATERIALS AND METHODS: Between 1999 and 2007, we collected data on 59 patients (39 male and 20 female patients) who received surgical intervention for acute lower intestinal hemorrhage requiring transfusion at our university clinic. Treatment complications and mortality were analyzed retrospectively. RESULTS: The average age of the patients in this study is 70.0 +/- 12.2 years (range, 39 to 97 years) with an overall mortality of 15.3%. Blood transfusions >10 U (p = 0.031), postoperative need for ventilation (p = 0.004), necessary reoperations (p = 0.016), and an initial hemoglobin level <80 g/L (p = 0.043) proved to be significant risk factors for death. Blood transfusions >10 U (p = 0.028), necessary reoperations (p = 0.001), and an initial hemoglobin level <80 g/L (p = 0.033) were found to be significant risk factors for postoperative complications. All other parameters have no significant impact. CONCLUSIONS: The decisive factors for the outcome of lower gastrointestinal hemorrhage requiring surgery are the severity of bleeding, beginning of treatment (initial hemoglobin level, need for packed red blood cells), and treatment efficiency (necessary reoperation). PMID- 19294393 TI - Measurements of the quadriceps femoris function after meniscus refixation at the stable athlete's knee. AB - INTRODUCTION: The purpose of this study was to monitor the muscular changes regarding the isokinetic strength and torque pattern of the quadriceps femoris at the stable athlete's knee after meniscus tear refixation. MATERIALS AND METHODS: Therefore 15 athletes (10 male, 5 female) performing recreational or competitional sports at least five times a week before injury were retrospectively examined in the average 2.5 years after isolated arthroscopic meniscus refixation using Inside Out technique. Next to function and sport activity focused scores the isokinetic peak torque (PT) and in the EMG have been analyzed compared to the uninjured knee. RESULTS: The mean age was 31.26 years. The time between injury and surgery was in the average 13.7 days. According to our first results the data suggest a complete recovery of functional and muscular pattern after meniscus refixation at the stable athlete's knee. No significant EMG changes for quadriceps femoris were detectable. The PT was fully recovered. The functional and sport activity score analysis (Lysholm and Tegner score) showed no changes in the postoperative long-term follow up compared to the preinjured status. CONCLUSION: Examining isokinetic PT and the EMG of the quadriceps femoris, these data show no side-to-side differences. Regarding the function and sports activity score system, the functionally high demand patients seem to profit by this procedure. PMID- 19294394 TI - CD4+CD25+ Treg cells in patients with toenail onychomycosis. AB - The essential role played by CD25+CD4+ regulatory T (Treg) cells in the control of immunity against some pathogens such as Helicobacter pylori is now well established. But their role in cutaneous fungal infections is still unknown. Onychomycosis is the chronic fungal infection of the nails, which is very common. The aim of this study was to evaluate possible relationship of CD4+CD25+ Treg cells and onychomycosis. Peripheral blood samples were investigated for CD4+CD25+ Treg cells using flow cytometry analysis in 43 toenail onychomycosis patients and in 30 healthy controls. We have found that onychomycosis patients had a higher expression of CD25+CD4+ Treg cells than controls, with values of 8.45 +/- 4.47% versus 4.64 +/- 1.59%, respectively (P = 0.001). The results of this study suggests that increased numbers of CD4+CD25+ Treg cells may play a role in failure of clearance of dermatophytes from skin by preventing the protective inflammation which is leading to development of onychomycosis. Accordingly, we address the possibility that CD4+CD25+ Treg cells may play a role in immune pathogenesis of other superficial fungal infections. PMID- 19294395 TI - Isolation of Leishmania amastigote protein fractions which induced lymphocyte stimulation and remission of psoriasis. AB - A first generation polyvalent vaccine (AS100(1)) was manufactured with protein from several cultured leishmania species, which proved to be effective in the treatment of psoriasis. To determine the effective factor, a single blind trial with four monovalent second generation vaccines (AS100(2)) was done in 26 subjects, which also resulted in remission of psoriasis. AS100(2) vaccines were further purified, resulting in seven chromatography fractions (AS200) per species. In vitro testing of the fractions on blood lymphocytes resulted in subjects being categorized as low or high responders before treatment. Both responder groups had no statistical difference in clinical outcome after AS100(1) treatment. Subsequently, a single-blind trial in 55 subjects treated with AS200 fractions from Leishmania brasiliensis also induced remission of Psoriasis. Two HIV +/- subjects with plaque psoriasis experienced remission after treatment with AS100(1). There are factors in leishmania species which induce remission of psoriasis by stimulating lymphocytes. PMID- 19294396 TI - Both all-trans retinoic acid and cytochrome P450 (CYP26) inhibitors affect the expression of vitamin A metabolizing enzymes and retinoid biomarkers in organotypic epidermis. AB - The biosynthesis of retinoic acid (RA) from retinol is controlled by several enzymes, e.g. dehydrogenases (RalDH2, RoDH-4) and retinol-esterifying enzyme (LRAT), whereas its degradation mainly involves CYP26 enzymes. In keratinocytes, RA activates the nuclear retinoid-receptors inducing the transcription of many genes. Here, we examined the effects of RA and the CYP26 inhibitors, liarozole and talarozole, on retinoid metabolism and RA-regulated genes in organotypic epidermis. RA induced the expression of CYP26 enzymes already after 8 h, whereas LRAT exhibited a later response and peaked at 48 h, indicating a feedback induction of retinol esterification. In line with a reduced biosynthesis of RA from retinol after exogenous RA, the expression of RDH16 reduced 80% in response to exogenous RA. The mRNA expression of RA-regulated genes (KRT2, KRT4, CRABPII and HBEGF) was altered within 24 h after RA exposure. In contrast, the CYP26 inhibitors caused only minor effects, except for a clear-cut induction of CYP26A1 only when combined with minute amounts of exogenous RA. Cellular accumulation of exogenous [3H]RA was higher after talarozole than after liarozole, probably indicating a greater CYP26-inhibitory potency of the former drug. The present study shows that CYP26A1 expression is extremely sensitive to both exogenous RA and increased endogenous RA levels, i.e. due to CYP26 inhibition, and thus an excellent biomarker for retinoid signalling in organotypic epidermis. PMID- 19294397 TI - Surgery and cancer after endometrial resection. Long-term follow-up on menstrual bleeding and hormone treatment by questionnaire and registry. AB - PURPOSE: Long-term follow-up 11 years after endometrial ablation. METHODS: A questionnaire on menstruation, hormone treatment, and operations on women, which had 421 endometrial ablations during 1990-1996, was added to registered data on cancer, operational and obstetrical history. RESULTS: Eighty-two percent of the questionnaires were returned and answered. Of those women on hormone treatment after ablation 26% had a hysterectomy later on compared to 34% of women with no hormone treatment after ablation (P = 0.28; chi (2) test). Hysterectomy was directly proportional to the amount of meno/metrorrhagia; however, the number of women with a hysterectomy was evenly distributed in relation to the duration of hormone treatment. The time of hormone treatment in relation to the ablation was not associated with the distribution of meno/metrorrhagia or whether the women had a hysterectomy later on. We observed three women with incidental endometrial cancer at follow-up and the expected numbers were 6.8 cases. CONCLUSION: Endometrial ablation is an effective form of treatment for menorrhagia with no increase in the incidence of endometrial cancer. Hormonal treatment does not seem to have any influence on the course of events. PMID- 19294398 TI - Case report of a sarcoma botryoides of the uterine cervix in fertile age and literature review. AB - INTRODUCTION: The sarcoma botryoides of the cervix uteri occurs very rarely in fertile women. CASE: Here is a case of a 35-year-old fertile woman, who developed a sarcoma botryoides of the cervix uteri. The diagnosis required clinical examination, ecography, NMR, CT scan and a pathological examination of the surgical sample. Radical surgery and chemotherapy was performed. The patients is still living after more than 1 year after treatment. DISCUSSION: Although the combined modality approach has given, if not constant, acceptable outcomes, a larger investigation on all the cases of sarcoma botryoides of the cervix uteri is expected. PMID- 19294399 TI - Cerebral plasticity in acute vestibular deficit. AB - The aim of this study was to analyze the effect of acute vestibular deficit on the cerebral cortex and its correlation with clinical signs and symptoms. Eight right-handed patients affected by vestibular neuritis, a purely peripheral vestibular lesion, underwent two brain single photon emission computed tomography (SPECT) in 1 month. The first SPECT analysis revealed reduced blood flow in the temporal frontal area of the right hemisphere in seven of eight patients, independent of the right/left location of the lesion. The alteration was present always in the right, non-dominant hemisphere and was reversible in some patients 1 month after the onset, together with attenuation of signs and symptoms. It may be hypothesized that the transient reduction of cortical blood flow and subsequently of cortical activity in the non-dominant hemisphere, also the expression of cerebral plasticity, may serve as a defense mechanism aimed to attenuate the vertigo symptom. PMID- 19294400 TI - 'Aggressive papillary' thyroid microcarcinoma. AB - The objective of this study is to highlight the fact that papillary thyroid microcarcinoma can be aggressive, requiring therapeutic management similar to that of other differentiated thyroid cancers. This 8-year retrospective study concerned 187 surgical patients managed in an ENT and Head and Neck surgery department for thyroid cancer. 65 patients were found to have papillary microcarcinoma. 41 microcarcinomas were considered to be aggressive because of the presence of several risk factors such as larger than 5 mm, multifocal microcarcinomas, capsular effraction, vascular embolus, tumour extension beyond the thyroid parenchyma and metastatic lymphadenopathy. All patients with aggressive papillary microcarcinoma were treated by total thyroidectomy and (131)I. Ipsilateral recurrent laryngeal and lateral cervical lymph node dissections were performed in ten patients, ipsilateral cervical lymph node dissection was performed in six patients and bilateral recurrent laryngeal and lateral cervical lymph node dissections were performed in three patients. No recurrence or metastasis was observed (follow-up ranging from 6 months to 8 years). The optimal management of thyroid papillary microcarcinoma is still controversial. "Aggressive" papillary thyroid microcarcinoma is not rare and may justify aggressive treatment depending on the presence or absence of prognostic risk factors. PMID- 19294401 TI - Extensive papillary carcinoma of the thyroid gland treated by external beam radiotherapy, surgery and postoperative radioactive iodine therapy. AB - The treatment of choice for well-differentiated thyroid carcinoma is surgery, radioactive iodine ablation and thyroid stimulating hormone suppression [Richard in Cancer 82:375-388, 1988; James et al. in Endocrinol Metab Clin North Am 37(2):497-509]. We report a case with an extensive papillary carcinoma of thyroid gland treated by external beam radiotherapy as a cytoreductive therapy followed by total thyroidectomy. Postoperatively the patient was given radioactive iodine. The patient is free from tumor a year after the treatment. PMID- 19294402 TI - Geometric morphometric methods for three-dimensional virtual reconstruction of a fragmented cranium: the case of Angelo Poliziano. AB - The process of forensic identification of missing individuals is frequently reliant on the superimposition of cranial remains onto an individual's picture and/or facial reconstruction. In the latter, the integrity of the skull or a cranium is an important factor in successful identification. Here, we recommend the usage of computerized virtual reconstruction and geometric morphometrics for the purposes of individual reconstruction and identification in forensics. We apply these methods to reconstruct a complete cranium from facial remains that allegedly belong to the famous Italian humanist of the fifteenth century, Angelo Poliziano (1454-1494). Raw data was obtained by computed tomography scans of the Poliziano face and a complete reference skull of a 37-year-old Italian male. Given that the amount of distortion of the facial remains is unknown, two reconstructions are proposed: The first calculates the average shape between the original and its reflection, and the second discards the less preserved left side of the cranium under the assumption that there is no deformation on the right. Both reconstructions perform well in the superimposition with the original preserved facial surface in a virtual environment. The reconstruction by means of averaging between the original and reflection yielded better results during the superimposition with portraits of Poliziano. We argue that the combination of computerized virtual reconstruction and geometric morphometric methods offers a number of advantages over traditional plastic reconstruction, among which are speed, reproducibility, easiness of manipulation when superimposing with pictures in virtual environment, and assumptions control. PMID- 19294403 TI - Fatal mycotic aneurysm caused by Pseudallescheria boydii after near drowning. AB - Pseudallescheria boydii is a rare cause of mycotic infection. Factors predisposing to systemic infection include traumas, immunosuppression, and near drowning. We report a case of multiple aneurysms caused by this hyalohyphomycete, occurred after near drowning. The car driven by a 53-year-old woman plunged into a canal in The Netherlands. After a 20-min-long submersion, the woman was rescued. At hospital, a severe aspiration of muddy water and a mycotic pneumonia were diagnosed. Despite the immediate prescription of a long-term antimycotic therapy and the initial good response, the patient died 4 months later. The autopsy showed a mycotic aneurysm of the abdominal aorta and multiple ruptured mycotic aneurysms of the circle of Willis with fatal subarachnoid bleeding. PMID- 19294404 TI - Evaluation of the ossification of the medial clavicular epiphysis with a digital ultrasonic system to determine the age threshold of 21 years. AB - In this study, both medial clavicular epiphyses of 77 men were evaluated by two researchers with a digital ultrasound system. The degree of ossification was classified in four stages according to Schulz et al. (Int J Legal Med 122:55-58, 17). Of the test persons over 21 years of age, stage 4 was found bilaterally in 25 (60%), while for five (12%) stage 4 was found on only one side; 12 (28%) further subjects did not show stage 4 on either side, although nine were exactly 21 years old. For the subjects under 21 years of age, stage 4 was found bilaterally in three cases (8.5%), and in three additional cases stage 4 was found on one side. Particularly, the three cases in which stage 4 was diagnosed bilaterally before completion of the 21st year of age limit the value of the used examinatory technique. For these cases it would be interesting to know if parts of the epiphyseal plate that might have been obstructed from the view of the ultrasound transducer would have been visible with CT. PMID- 19294405 TI - Effect of vitrectomy on macular microcirculation in patients with diffuse diabetic macular edema. AB - PURPOSE: To determine whether retinal microcirculatory changes occur after vitrectomy in eyes with diffuse diabetic macular edema (DME), and whether changes in blood flow are associated with visual outcome and the resolution of macular edema. METHODS: Thirty-three eyes of 30 consecutive diabetic patients who underwent pars plana vitrectomy for diffuse DME, and 16 eyes of 16 diabetic patients without macular edema, were included. Mean macular blood flows were measured using a Heidelberg Retinal Flowmeter, and central macular thicknesses (CMTs) were determined by optical coherence tomography. Visual outcomes, CMTs, and macular blood flow were evaluated before and 1, 4, 12 weeks after vitrectomy, and compared between eyes with resolved macular edema and those with persistent macular edema. RESULTS: Mean preoperative macular blood flow in eyes with diffuse DME was higher than in controls (606.5+/-357.9 AU vs 407.1+/-265.9 AU, P=0.021). Mean macular blood flow (422.6+/-247.5 AU) at 12 weeks after vitrectomy was significantly lower than preoperative blood flow (P=0.002), and similar to that of controls (P=0.47). In 22 of the 33 (66.7%) DME eyes, macular edema was resolved at 12 weeks after vitrectomy. The mean ratio of macular blood flow at 12 weeks postoperatively versus the preoperative level was significantly lower in eyes with resolved macular edema than in eyes with persistent macular edema (0.65 vs 1.08, P<0.001). CONCLUSIONS: Increased macular blood flow in diabetic macular edema was normalized after vitrectomy in eyes with resolved macular edema. Changes of macular blood flow may be associated with the resolution of macular edema in diabetic eyes. PMID- 19294406 TI - An active mechanism flanks and modulates the export of the small ribosomal subunits. AB - The modalities of export of the ribosomal subunits from the nucleolus to the nuclear pores have been only partially clarified since it is not yet clear whether the movements depend purely on diffusion or also from an active process. Recently, we suggested the existence of an active transport mechanism of a subset (10-12%) of the small ribosomal subunits (SSU) (Cisterna et al. in 2006, Faseb J). Here, we give further evidence that an active, motor protein-mediated process exists for the SSU transport from the nucleolus to the nuclear pore. We demonstrate that the blockade of ATP synthesis and antibody-mediated inhibition of nuclear myosin or actin induce structural and functional modifications of the nucleolus, suggestive of transcriptional activity decrease. Moreover, both treatments induce a significant retention of RNA inside the nucleus and an accumulation of ribosomal subunits in the granular component. We suggest that the existence of this secondary, active mechanism of SSU transport might be utilized by the cell when a more rapid and directional export is needed. PMID- 19294407 TI - Expression and localization of growth hormone receptor in the oviduct of cyclic and pregnant pigs and mid-implantation conceptuses. AB - Previously it was shown that growth hormone (GH) and its receptor (GH-R) are involved in growth-promoting events during early embryonic development. However, it is still unknown if GH-induced GH-R signalling may support other functions within the oviduct. The purpose of our study was to analyse GH-R expression and localization in the porcine oviduct during different stages of the oestrus cycle and pregnancy (days 2-3 post inseminationem to days 65-71). As shown by reverse transcription polymerase chain reaction (RT-PCR), GH-R is expressed in the porcine oviduct during all stages of the oestrus cycle and pregnancy, respectively. Additionally, GH-R mRNA was detected in porcine conceptuses collected at day 18 of pregnancy. Using immunohistochemistry, GH-R was exclusively localized to the epithelium of the porcine oviduct throughout all segments examined. Localization of GH-R was mainly observed in the cytoplasm of ciliated epithelial cells. Generally, the number of GH-R-positive cells was elevated in the periovulatory phase of the oestrus cycle. Except for the isthmic epithelium, staining intensity of GH-R-positive cells was highest at oestrus and markedly reduced at met- and dioestrous stages. In infundibular and ampullar segments, percentage of GH-R-positive cells was significantly higher at days 2-3 post inseminationem compared to days 65-71 of pregnancy. Furthermore, in porcine conceptuses on day 18 of pregnancy GH-R protein expression was almost exclusively localized to trophectoderm. Our data suggest that GH-R mRNA and protein expression in the porcine oviduct throughout the oestrus cycle and pregnancy may suggest other activities of GH not described previously. Specifically, autocrine or paracrine GH-induced GH-R signalling may be linked to ciliated cell homeostasis of the porcine oviduct. Additionally, our results indicate that GH-R expression in the pig trophectoderm may be responsible for trophoblastic elongation. PMID- 19294408 TI - Mechano-transduction to muscle protein synthesis is modulated by FAK. AB - We examined the involvement of focal adhesion kinase (FAK) in mechano-regulated signalling to protein synthesis by combining muscle-targeted transgenesis with a physiological model for un- and reloading of hindlimbs. Transfections of mouse tibialis anterior muscle with a FAK expression construct increased FAK protein 1.6-fold versus empty transfection in the contralateral leg and elevated FAK concentration at the sarcolemma. Altered activation status of phosphotransfer enzymes and downstream translation factors showed that FAK overexpression was functionally important. FAK auto-phosphorylation on Y397 was enhanced between 1 and 6 h of reloading and preceded the activation of p70S6K after 24 h of reloading. Akt and translation initiation factors 4E-BP1 and 2A, which reside up- or downstream of p70S6K, respectively, showed no FAK-modulated regulation. The findings identify FAK as an upstream element of the mechano-sensory pathway of p70S6K activation whose Akt-independent regulation intervenes in control of muscle mass by mechanical stimuli in humans. PMID- 19294409 TI - Shoulder abduction torque steadiness is preserved in subacromial impingement syndrome. AB - This study compared peak torque and torque steadiness during isometric abduction in subjects with subacromial impingement syndrome (SIS) and those with no upper limb disorders. The SIS group consisted of 27 subjects (33.48 +/- 9.94 years) with unilateral SIS. The control group consisted of 23 healthy and active subjects (32.26 +/- 9.04 years). Peak torque and torque steadiness were measured during isometric abduction (80 masculine in the scapular plane) of the shoulder. Standard deviation, coefficient of variation, stability time, median frequency, and relative power were measured from the steadiness trials. There were neither significant interactions between group and side (P > 0.05), nor were there significant main effects of group and side (P > 0.05) for all variables analyzed. The results of this study showed that steadiness is preserved by SIS during isometric abduction of the shoulder. PMID- 19294410 TI - Physiological correlates of performance. Case study of a world-class rower. AB - This report describes the changes in physiological capacity of a heavy-weight rower who obtained seven medals in World Championships and Olympic Games. The investigation was carried out over the last 6 years of the rower's international competition career in comparison with peer champions, and the following 4 years. Over the first period, maximal oxygen uptake (VO(2max)) remained above 6 l min( 1) which is an outstanding value. The training load measured over the last 18 months of the period increased from 119 to 142 km wk(-1) of rowing. Four years after the international competition period, VO(2max) had only declined by 3.6% although the training load had declined by 35%. These data suggest that the ability of this rower to compete at top level for years was related to ability to maintain an outstanding VO(2max). Gross efficiency and ability to rely on anaerobic glycolysis did not emerge as relevant factors. PMID- 19294411 TI - Time course of haemoglobin mass during 21 days live high:train low simulated altitude. AB - The aim of this study was to determine the time course of changes in haemoglobin mass (Hb(mass)) in well-trained cyclists in response to live high:train low (LHTL). Twelve well-trained male cyclists participated in a 3-week LHTL protocol comprising 3,000 m simulated altitude for ~14 h/day. Prior to LHTL duplicate baseline measurements were made of Hb(mass), maximal oxygen consumption (VO(2max)) and serum erythropoietin (sEPO). Hb(mass) was measured weekly during LHTL and twice in the week thereafter. There was a 3.3% increase in Hb(mass) and no change in VO(2max) after LHTL. The mean Hb(mass) increased at a rate of ~1% per week and this was maintained in the week after cessation of LHTL. The sEPO concentration peaked after two nights of LHTL but there was only a trivial correlation (r = 0.04, P = 0.89) between the increase in sEPO and the increase in Hb(mass). Athletes seeking to gain erythropoietic benefits from moderate altitude need to spend >12 h/day in hypoxia. PMID- 19294412 TI - Postoperative hemorrhagic complications after elective laparoscopic cholecystectomy in patients receiving long-term anticoagulant therapy. AB - BACKGROUND AND AIMS: Among patients on long-term anticoagulant therapy who undergo laparoscopic cholecystectomy (LC), bleeding complications have not been extensively investigated. The objective of this study was therefore to investigate postoperative bleeding complications prospectively in patients on chronic oral anticoagulation. MATERIALS AND METHODS: In the period of January 2002 to December 2007, 44 patients on long-term anticoagulation with warfarin, an oral anticoagulant (OAC), underwent LC in our center. Oral anticoagulant was discontinued 5 days before the planned date of surgery, and patients were admitted to the hospital 3 days before. Upon admission, bridging anticoagulation with enoxaparin, a low molecular weight heparin (LMWH), was started. When their international normalized ratio (INR) decreased to <1.5, patients underwent LC. In the absence of postoperative bleeding complications, OAC and LMWH were resumed on the evening of the day of surgery, and LMWH was continued until each patient's target INR was reached. A comparison group was comprised by 1,421 consecutively enrolled patients with no comorbid disease who underwent LC during the same period. RESULTS: In the comparison group, postoperative bleeding was encountered in 21 patients (1.5%). In the anticoagulation group, postoperative bleeding was encountered in 11 patients (25%) and ranged from minor oozing from a port incision in one patient to hemorrhage, sepsis, and fatality in one patient. In the anticoagulation group, no significant differences were found between patients with and without postoperative bleeding in terms of age, gender, body mass index, American Society of Anesthesiologists score, INR, or other hemostasis parameters. CONCLUSIONS: In patients who underwent LC with bridging anticoagulation, postoperative bleeding was markedly more frequent than expected and was not predicted by the usual coagulation parameters. This suggests a need for methods that can indicate which patients on long-term anticoagulation are at risk for postoperative bleeding. PMID- 19294413 TI - A molecularly guided tour along the nephron. PMID- 19294414 TI - RhoA-ROCK signaling is involved in contraction-mediated inhibition of SERCA2a expression in cardiomyocytes. AB - In neonatal ventricular cardiomyocytes (NVCM), decreased contractile activity stimulates sarco-endoplasmic reticulum Ca(2+)-ATPase2a (SERCA2a), analogous to reduced myocardial load in vivo. This study investigated in contracting NVCM the role of load-dependent RhoA-ROCK signaling in SERCA2a regulation. Contractile arrest of NVCM resulted in low peri-nuclear localized RhoA levels relative to contracting NVCM. In arrested NVCM, ROCK activity was decreased (59%) and paralleled a loss in F-actin levels. Y-27632-induced ROCK inhibition in contracting NVCM increased SERCA2a messenger RNA expression by 150%. This stimulation was transcriptional, as evident from transfections with the SERCA2a promoter. A reciprocal effect of Y-27632 treatment on the promoter activity of atrial natriuretic factor was observed. SERCA2a transcription was not altered by co-transfection of the RhoA-ROCK-dependent serum response factor (SRF) alone or in combination with myocardin. Furthermore, GATA4, another ROCK-dependent transcription factor, induced rather than repressed SERCA2a transcription. This study shows that contractile activity suppresses SERCA2a gene expression via RhoA ROCK-dependent transcription modulation. This modulation is likely to be accomplished by a transcription factor other than SRF, myocardin, or GATA4. PMID- 19294415 TI - Nitric oxide donor-mediated inhibition of phosphorylation shows that light mediated degradation of photosystem II D1 protein and phosphorylation are not tightly linked. AB - An outcome of the photochemistry during oxygenic photosynthesis is the rapid turn over of the D1 protein in the light compared to the other proteins of the photosystem II (PS II) reaction center. D1 is a major factor of PS II instability and its replacement a primary event of the PS II repair cycle. D1 also undergoes redox-dependent phosphorylation prior to its degradation. Although it has been suggested that phosphorylation modulates D1 metabolism, reversible D1 phosphorylation was reported not to be essential for PS II repair in Arabidopsis. Thus, the involvement of phosphorylation in D1 degradation is controversial. We show here that nitric oxide donors inhibit in vivo phosphorylation of the D1 protein in Spirodela without inhibiting degradation of the protein. Thus, D1 phosphorylation is not tightly linked to D1 degradation in the intact plant. PMID- 19294416 TI - Status of HER1 and HER2 in peritoneal, ovarian and colorectal endometriosis and ovarian endometrioid adenocarcinoma. AB - A role for the EGF system, in particular HER1 and 2, in growth of the endometrium has been suggested but HER1 and 2 have not been studied in all locations of endometriosis and in ovarian endometrioid adenocarcinoma (OEC) which is a rare form of malignant transformation of endometriosis. Immunohistochemistry (IHC) was used for studying HER1 and HER2 in ovarian (n = 10), peritoneal (n = 10), colorectal endometriosis (n = 20) and OEC (n = 10). Fluorescent in situ hybridisation (FISH) was used for analysing the status of HER2 gene in colorectal endometriosis and OEC. All samples were negative for HER2 in both glandular and stromal cells and in glandular cells for HER1 by IHC. In 15 out of 20 colorectal endometriosis, there was a weak expression in stromal cells. Following FISH, two colorectal samples had a partial 17 aneusomy and three OEC, a 17 polysomy. The other samples were 17 disomic without HER2 amplification; HER1 and 2 do not seem to have a role in endometriosis physiopathology. PMID- 19294417 TI - EpCAM is overexpressed in gastric cancer and its downregulation suppresses proliferation of gastric cancer. AB - PURPOSE: To identify the role of epithelial cellular adhesion molecule (EpCAM) in gastric cancer growth and explore the potential value of EpCAM monoclonal antibody as new therapeutic strategy for gastric cancer. METHODS: The expression of EpCAM was determined by immunohistochemistry staining in gastric cancer tissues, RT-PCR and Western blot in cell lines. EpCAM expression in cell lines was downregulated by small interfering RNA. Then the effects of EpCAM on gastric cancer cell growth in vivo and in vitro were determined by MTT, FCM analysis, clone formation assay and tumor formation assay. Additionally, western blot was used to detect the effect of EpCAM on cell cycle-relevant factor cyclin D1. RESULTS: EpCAM was found to be overexpressed in gastric cancer tissues and cell lines. Downregulation of EpCAM resulted in a decrease of cell proliferation and cell cycle arrest in AGS and SGC7901 cells, which had high endogenous EpCAM expression. EpCAM downregulation also suppressed tumor formation in nude mice. Moreover, EpCAM repression in gastric cancer cells could downregulate cyclin D1. CONCLUSIONS: EpCAM was a potential oncogene and contributed to the growth of gastric cancer. Our data first provided compelling evidence of potential value of EpCAM in the therapy of gastric cancer in clinic. PMID- 19294418 TI - Intrahepatic cholangiocarcinoma: report of 272 patients compared with 5,829 patients with hepatocellular carcinoma. AB - PURPOSE: To clarify clinicopathologic differences between patients with intrahepatic cholangiocarcinoma (ICC) and hepatocellular carcinoma (HCC), and identify potential factors influencing survival after hepatectomy for ICC. METHODS: Comparison of clinicopathologic data was made between patients who underwent hepatectomy for ICC (n = 272) and HCC (n = 5,829) during the same period. Twenty-five clinicopathologic variables were selected for univariate and multivariate analyses to evaluate their influence on prognosis of ICC. RESULTS: Compared with patients with HCC, ICC patients were more common in females and more elderly, had a lower proportion of asymptomatic tumors, lower serum alpha fetoprotein, higher serum carcinoembryonic antigen, carbohydrate antigen 19-9 and alkaline phosphatase levels; lower incidence of hepatitis history, associated cirrhosis and serum hepatitis B surface antigen; lower proportion of small tumors, well-encapsulated tumors and tumor emboli in the portal vein; higher proportion of single tumor, perihila lymph node involvement and poor differentiation; and less frequency of limited resection (all, P < 0.0001). Distant metastasis was less frequent in patients with ICC (P = 0.027). A total of 5-years overall and disease-free survival (in brackets) after resection was 26.4% (13.1%) and 44.5% (33.1%) (P < 0.0001, P < 0.0001) for patients with ICC and HCC, respectively. Factors influencing survival after resection of ICC can be divided mainly into two categories: early detection of asymptomatic ICC (P < 0.0001) and curative resection (P = 0.002). CONCLUSION: ICC Patients have distinct clinicopathologic features as compared with HCC patients. Surgery remains the only effective treatment for ICC. Early detection of asymptomatic ICC and curative resection were the key to achieve optimal survival. PMID- 19294419 TI - Twenty-year survivors after resection for hepatocellular carcinoma-analysis of 53 cases. AB - PURPOSE: To clarify the clinicopathologic features of patients surviving > or =20 years after resection for hepatocellular carcinoma (HCC). METHODS: Between 1961 and 1987, a total of 396 patients underwent hepatic resection for HCC; 53 (13.4%) patients survived > or =20 years, and 343 (86.6%) patients survived <20 years. A comparative study between the two groups was made. RESULTS: By March of 2007, 67.6% (36/53) patients are still alive, disease free; 5.7% (3/53) patients died of tumor recurrence or metastasis; 11.3% (6/53) patients died of liver failure; 5.7% (5/53) patients were lost during follow-up. The longest patient survived 43 years and 2 months. Five young patients got married after resection and have had babies. One patient with a tumor measuring 17 x 13 x 9 cm (largest tumor in this series) survived for 37 years after resection, still alive, free of disease. Reresection for recurrence was done in nine patients, mean survival being 26 years and 11 months. Reresection for solitary pulmonary metastasis was carried out in three patients, mean survival being 29 years and 2 months. In comparison with patients surviving <20 years, patients surviving > or =20 years were significantly younger (P = 0.031), had a higher incidence of asymptomatic tumors (56.6 vs. 34.4%, P = 0.002); lower gamma-glutamyl transpeptidase level (< or =50 U/L, 64.2 vs. 25.9%, P < 0.000), lower proportion of liver cirrhosis (66.0 vs. 83.6%, P = 0.002); higher percentage of small tumors (< or =5 cm, 62.3 vs. 29.9%, P < 0.000), single nodule tumors (90.6 vs. 62.9%, P < 0.000), and well encapsulated tumors (86.8 vs. 43.6%, P < 0.000); lower proportion of tumor emboli in the portal vein (3.8 vs. 22.5%, P = 0.002), better differentiation of tumor cells (Edmondson grade I, 21.6 vs. 9.1%, P = 0.036), and higher curative resection rate (100 vs. 64.1%, P < 0.000). CONCLUSIONS: Early detection and curative resection are the principal factors improving long-term survival. Long term follow-up after resection of HCC is very important, and should continue for the remainder of the patient's life. Reresection for recurrence and metastasis is important approach to improve prognosis. PMID- 19294420 TI - Return to sender: use of Plasmodium ER retrieval sequences to study protein transport in the infected erythrocyte and predict putative ER protein families. AB - We have investigated how knowledge of endoplasmic reticulum (ER) retrieval signals can be used to study specific trafficking pathways in the malaria infected erythrocyte. We show that addition of various lumenal ER retrieval signals to soluble green fluorescent protein (GFP) chimaera causes retrieval of the fusion protein in the parasite's ER. In contrast, adding these signals to the C-terminus of a membrane bound protein does not affect its eventual sub-cellular localization. This demonstrates proof of principle that ER retrieval signals can be used to study the solubility state of Plasmodium falciparum proteins during their transport to the host erythrocyte. Furthermore, using our knowledge of ER retrieval signals, we identify Plasmodium ER protein families and assign putative functions to them. PMID- 19294421 TI - Evaluation of Babesia bigemina 200 kDa recombinant antigen in enzyme-linked immunosorbent assay. AB - A truncated fragment of the gene encoding the 200-kDa protein (P200) of Babesia bigemina was cloned into a plasmid vector, pGEX-4 T-1 and expressed in Escherichia coli as a glutathione-S-transferase fused protein. An indirect enzyme linked immunosorbent assay (ELISA) using the rp200/CT detected specific antibodies in cattle experimentally infected with B. bigemina. Furthermore, the antigen did not cross-react with antibodies to Babesia bovis, a closely related Babesia parasite indicating that rp200/CT is a specific antigen for the diagnosis of B. bigemina infection. Additionally, ELISA using p200/CT and polymerase chain reaction were conducted on serum and corresponding DNA samples obtained from field cattle to evaluate the diagnostic utility of the p200/CT antigen. Results from the current study suggest that p200/CT ELISA is a sensitive and specific method for improved serodiagnosis of B. bigemina infection. PMID- 19294422 TI - Fast high yield of pure Leishmania (Leishmania) infantum axenic amastigotes and their infectivity to mouse macrophages. AB - Leishmania (L.) infantum (syn. Leishmania chagasi) is a dimorphic protozoan parasite that lives in promastigote and amastigote form in its sandfly vector and mammalian hosts, respectively. Here, we describe an in vitro culture system for the generation of a pure population of L. infantum axenic amastigotes after only 4 days incubation in culture medium supplemented with fetal calf serum, human urine, L: -glutamine, and HEPES at 37 masculineC (pH 5.5). Ultrastrutural analysis and infection assays in two macrophage populations (Kupffer cells (KUP) and peritoneal macrophages (PM)) infected with axenic amastigotes demonstrated that they maintained morphological and biochemical (A2 expression) features and a similar infection pattern to tissue-derived L. infantum amastigotes. The susceptibility of the macrophage lines to axenic or tissue-derived amastigotes and promastigotes was investigated. We found a completely different susceptibility profile for KUP and PM. Liver macrophages, both KUP and immigrant macrophages, are intimately involved in the response to L. infantum infection; this difference in susceptibility is probably related to their capacity to eliminate these parasites. Our in vitro system was thus able to generate axenic amastigotes that resemble tissue-derived amastigotes both in morphology and infectivity pattern; this will help in further investigation of the biological characteristics of the host-parasite relationship as well as the process of pathogenesis. PMID- 19294424 TI - Genetic foundations of human intelligence. AB - Individual differences in intelligence (cognitive abilities) are a prominent aspect of human psychology, and play a substantial role in influencing important life outcomes. Their phenotypic structure-as described by the science of psychometrics-is well understood and well replicated. Approximately half of the variance in a broad range of cognitive abilities is accounted by a general cognitive factor (g), small proportions of cognitive variance are caused by separable broad domains of mental function, and the substantial remainder is caused by variance that is unique to highly specific cognitive skills. The heritability of g is substantial. It increases from a low value in early childhood of about 30%, to well over 50% in adulthood, which continues into old age. Despite this, there is still almost no replicated evidence concerning the individual genes, which have variants that contribute to intelligence differences. Here, we describe the human intelligence phenotype, summarise the evidence for its heritability, provide an overview of and comment on molecular genetic studies, and comment on future progress in the field. PMID- 19294425 TI - The value of sequential dialysis, mannitol and midodrine in managing children prone to dialysis failure. AB - The uremic state impairs compensatory responses to ultrafiltration (UF). Intradialytic symptoms and hypotension can result and lead to premature discontinuation of treatment and sub-optimal dialysis. We report the benefits of mannitol, sequential dialysis and midodrine in reducing dialysis failures in those children prone to intradialytic hypotension. Prophylactic mannitol halved the odds of intradialytic symptoms and hypotension and increased UF volumes. Sequential dialysis halved the odds of symptoms but hypotension persisted. In one patient with refractory hypotension, only intradialytic midodrine consistently maintained acceptable intradialytic blood pressures, reduced intradialytic symptoms and increased the UF potential. PMID- 19294426 TI - Intellectual and motor performance, quality of life and psychosocial adjustment in children with cystinosis. AB - Cystinosis is a rare multisystemic progressive disorder mandating lifelong medical treatment. Knowledge on the intellectual and motor functioning, health related quality of life and psychosocial adjustment in children with cystinosis is limited. We have investigated nine patients (four after renal transplantation) at a median age of 9.7 years (range 5.3-19.9 years). Intellectual performance (IP) was analysed with the Wechsler Intelligence Scale for Children-III (seven children) and the Kaufman Assessment Battery for Children (two children). Motor performance (MP) was evaluated using the Zurich Neuromotor Assessment Test, and quality of life (QOL) was studied by means of the Netherlands Organization for Applied Scientific Research Academical Medical Center Child Quality of Life Questionnaire. Psychosocial adjustment was assessed by the Child Behavior Checklist. The overall intelligence quotient (IQ) of our patient cohort (median 92, range 71-105) was significantly lower than that of the healthy controls (p = 0.04), with two patients having an IQ < 85. Verbal IQ (93, range 76-118) was significantly higher than performance IQ (90, range 68-97; p = 0.03). The MP was significantly below the norm for pure motor, pegboard and static balance, as well as for movement quality. The patients' QOL was normal for six of seven dimensions (exception being positive emotions), whereas parents reported significant impairment in positive emotions, autonomy, social and cognitive functions. Significant disturbance was noted in terms of psychosocial adjustment. Based on the results from our small patient cohort, we conclude that intellectual and motor performance, health-related QOL and psychosocial adjustment are significantly impaired in children and adolescents with cystinosis. PMID- 19294427 TI - Wilms tumor arising in a child with X-linked nephrogenic diabetes insipidus. AB - We report on a child with X-linked nephrogenic diabetes insipidus (NDI) who developed Wilms tumor (WT). Nephrogenic diabetes insipidus is caused by mutations of the arginine vasopressin receptor (AVPR2) or aquaporin-II (AQP2) genes. Wilms tumor is also genetically heterogeneous and is associated with mutations of WT1 (15-20%), WTX (20-30%) and other loci. The boy presented at 5 months with failure to thrive, polyuria, hypernatremia and abdominal mass. Analysis of leukocyte DNA showed a novel missense mutation (Q174H) of the AVPR2 gene, which was not present in his mother. In cells (WitS) isolated from the tumor, WTX mRNA expression and coding sequence were intact. However, we identified a 44-kb homozygous deletion of the WT1 gene spanning exons 4 to 10. The WT1 deletion was not present in leukocyte DNA from the patient or his mother. We also noted strong beta-catenin (CTNNB1) expression in the tumor cells and identified a heterozygote missense Ser45Cys mutation of exon 3 of CTNNB1. However, the mutation was absent both in the constitutional DNA of the patient and his mother. The concurrence of WT and NDI has not been previously reported and may be unrelated. Nevertheless, this case nicely illustrates the sequence of events leading to sporadic Wilms tumor. PMID- 19294428 TI - Hyponatremia in children with respiratory tract infection. PMID- 19294429 TI - Single dose of palonosetron plus dexamethasone to control nausea, vomiting and to warrant an adequate food intake in patients treated with highly emetogenic chemotherapy (HEC). Preliminary results. AB - INTRODUCTION: The aim of our study was to evaluate the efficacy of a single-dose palonosetron plus dexamethasone to control emesis in patients (pts) receiving HEC. Moreover, we evaluated the amount of their food intake (FI) in the week following therapy, in order to measure any reduction of calories consumption related to Chemotherapy-induced nausea and vomiting (CINV). METHODS: Patients affected with advanced cancer were treated with palonosetron 250 mcg plus dexamethasone 20 mg before HEC. Nausea, vomiting, and FI were monitored by a 7 day diary. Complete Response (CR: no vomiting and no rescue therapy) was the primary endpoint, Complete Control (CC: CR and no more than mild nausea) and the evaluation of FI were the secondary endpoints. The endpoints were evaluated during the acute (0-24 h), the delayed (25-168 h) and overall (0-168 h) phases. RESULTS: Thirty-five patients were enrolled; 85.7% and 82.9% of patients achieved CR and CC respectively, during the acute phase; 82.9% and 77.1% of patients achieved CR and CC, during the delayed phase; 80% and 77.1% of patients achieved CR and CC, during the overall phase. During the acute phase, patients with a CC without nausea had a median daily FI of 1,575 kcal, whereas patients with CC and presence of mild nausea had a median daily FI of 1,040 kcal (-535 kcal; p < 0.0001). CONCLUSIONS: Our preliminary results confirm the efficacy of a single dose palonosetron plus dexamethasone to prevent both acute and delayed nausea and vomiting. Moreover, the efficacy of palonosetron in nausea and vomiting control seems to warrant adequate caloric intake in these patients. PMID- 19294430 TI - Diagnosis and surgical treatment of back pain originating from endplate. AB - Twenty-one patients with back pain originating from the endplate injuries were selected to explore the methods of diagnosis and surgical treatment. All patients underwent examinations using radiography, CT, and MR imaging. Pain level of disc was determined through discography in each patient. The principal outcome judgments were pain and disability, and the efficacy of surgical treatment was assessed through the use of the visual analog scale (VAS) for pain and the Oswestry disability index (ODI) for functional recovery. All 21 patients with a diagnosis of back pain originating from endplate injuries according to discography were treated with anterior or posterior fusion surgery. The mean follow-up period was 3 years and 5 months (range, 2-6 years). Of the 21 patients, 20 (20/21) reported a disappearance or marked alleviation of low back pain and experienced a definite improvement in physical function. Statistically significant and clinically meaningful improvements in the VAS and ODI scores were obtained after treatment in the patients with chronic low back pain originating from the endplate injuries (P = 0.0001). The study suggests that discography and fusion surgery may be very effective methods for the diagnosis and treatment, respectively, of chronic back pain originating from the endplate injuries. PMID- 19294431 TI - Arteriovenous fistula of the superior gluteal artery as a complication of posterior iliac crest bone graft harvesting: 3D-CT angiography and arterial embolization. AB - Superior gluteal artery injuries are rare, but potentially serious complications that occur during posterior iliac crest bone graft harvesting. The authors reported an arteriovenous fistula of the superior gluteal artery, which occurred as a complication during posterior iliac crest bone graft harvesting and was diagnosed with 3D-CT angiography, then treated with arterial embolization. PMID- 19294432 TI - A comparison of angled sagittal MRI and conventional MRI in the diagnosis of herniated disc and stenosis in the cervical foramen. AB - The object of this study is to demonstrate that angled sagittal magnetic resonance imaging (MRI) enables the precise diagnosis of herniated disc and stenosis in the cervical foramen, which is not available with conventional MRI. Due to both the anatomic features of the cervical foramen and the limitations of conventional MR techniques, it has been difficult to identify disease in the lateral aspects of the spinal canal and foramen using only conventional MRI. Angled sagittal MRI oriented perpendicular to the true course of the foramina facilitates the identification of the lateral disease. A review of 43 patients, who underwent anterior cervical discectomy and interbody fusion, is presented with a herniated disc and/or stenosis in the cervical foramen. They all had undergone conventional MRI and angled sagittal MRI. Fifty levels were surgically explored for evidence of foraminal herniated disc and stenosis. The results of each test were correlated with what was found at each explored surgical level. The sensitivity, specificity, and accuracy of both examinations for making the diagnosis of foraminal herniated disc and stenosis were compared. During the diagnosis of foraminal herniated disc, the sensitivity, specificity, and accuracy of angled sagittal MRI were 96.7, 95.0, and 96.0%, respectively, compared with 56.7, 85.0, and 68.0% for conventional MRI. In making the diagnosis of foraminal stenosis, the sensitivity, specificity, and accuracy of angled sagittal MRI were 96.3, 95.7, and 96.0%, respectively, compared with 40.7, 91.3, and 66.0% for conventional MRI. In the above groups, the difference between the tests for making the diagnosis of both foraminal herniated disc and stenosis was found to be statistically significant in sensitivity and accuracy. Angled sagittal MRI was a more accurate test compared to conventional MRI for making the diagnosis of herniated disc and stenosis in the cervical foramen. It can be utilized for the precise diagnosis of foraminal herniated disc and stenosis difficult or ambiguous in conventional MRI. PMID- 19294433 TI - Money matters: exploiting the data from outcomes research for quality improvement initiatives. AB - In recent years, there has been an increase in studies that have sought to identify predictors of treatment outcome and to examine the efficacy of surgical and non-surgical treatments. In addition to the scientific advancement associated with these studies per se, the hospitals and clinics where the studies are conducted may gain indirect financial benefit from participating in such projects as a result of the prestige derived from corporate social responsibility, a reputational lever used to reward such institutions. It is known that there is a positive association between corporate social performance and corporate financial performance. However, in addition to this, the research findings and the research staff can constitute resources from which the provider can reap a more direct benefit, by means of their contribution to quality control and improvement. Poor quality is costly. Patient satisfaction increases the chances that the patient will be a promoter of the provider to friends and colleagues. As such, involvement of the research staff in the improvement of the quality of care can ultimately result in economic revenue for the provider. The most advanced methodologies for continuous quality improvement (e.g., six-sigma) are data driven and use statistical tools similar to those utilized in the traditional research setting. Given that these methods rely on the application of the scientific process to quality improvement, researchers have the adequate skills and mind-set to embrace them and thereby contribute effectively to the quality team. The aim of this article is to demonstrate by means of real-life examples how to utilize the findings of outcome studies for quality management in a manner similar to that used in the business community. It also aims to stimulate research groups to better understand that, by adopting a different perspective, their studies can be an additional resource for the healthcare provider. The change in perspective should stimulate researchers to go beyond the traditional studies examining predictors of treatment outcome and to see things instead in terms of the "bigger picture", i.e., the improvement of the process outcome, the quality of the service. PMID- 19294434 TI - Electron transfer from cytochrome c to cupredoxins. AB - Electron transfer (ET) through and between proteins is a fundamental biological process. The activation energy for an ET reaction depends upon the Gibbs energy change upon ET (DeltaG(0)) and the reorganization energy. Here, we characterized ET from Pseudomonas aeruginosa cytochrome c(551) (PA) and its designed mutants to cupredoxins, Silene pratensis plastocyanin (PC) and Acidithiobacillus ferrooxidans rusticyanin (RC), through measurement of pseudo-first-order ET rate constants (k(obs)). The influence of the DeltaG (0) value for ET from PA to PC or RC on the k(obs) value was examined using a series of designed PA proteins exhibiting a variety of E (m) values, which afford the DeltaG (0) variation range of 58-399 meV. The plots of the k(obs) values obtained against the DeltaG(0) values for both PA-PC and PA-RC redox pairs could be fitted well with a single Marcus equation. We have shown that the ET activity of cytochrome c can be controlled by tuning the E(m) value of the protein through the substitution of amino acid residues located in hydrophobic-core regions relatively far from the redox center. These findings provide novel insights into the molecular design of cytochrome c, which could be utilized for controlling its ET activity by means of protein engineering. PMID- 19294435 TI - Influence of bisphosphonates on endothelial cells, fibroblasts, and osteogenic cells. AB - Bisphosphonate-associated osteonecrosis of the jaws (BP-ONJ) is a side effect primarily in patients receiving highly potent nitrogen-containing bisphosphonates. The exact etiopathology is unknown. In addition to reduced bone remodeling, there may also be an impact on soft tissues. The impact of nitrogen- (ibandronate, pamidronate, zoledronate) and non-nitrogen-containing bisphosphonates (clodronate) on human umbilicord vein endothelial cells (HUVEC), fibroblasts and osteogenic cells was analyzed employing cell viability testing and a scratch wound assay. The impact on the cell morphology of vital-stained osteogenic cells was investigated by cell visualization (confocal laser scanning microscopy). Pamidronate and zoledronate had the greatest negative impact on all cell lines, whereas the impact of ibandronate and clodronate was less distinct. The effect of clodronate on HUVEC and fibroblasts was particularly marginal. BP ONJ could be a multifactorial event with multicellular impairments. This might result in altered wound healing. The increased impact of the highly potent bisphosphonates, particularly on non-bone cells, may explain the higher occurrence of BP-ONJ. PMID- 19294436 TI - Two modifications in the treatment of keratocystic odontogenic tumors (KCOT) and the use of Carnoy's solution (CS)--a retrospective study lasting between 2 and 10 years. AB - This retrospective study aimed at evaluating the recurrence rates of keratocystic odontogenic tumors (KCOTs) that were enucleated with and without the application of Carnoy's solution (CS). The study included 36 KCOTs treated between 1996 and 2006. Recurrence rates were investigated in correlation with the respective treatment method applied. Additionally, any damage to the inferior alveolar nerve associated with treatment was analyzed. Treatments consisted of enucleation with (38.9%) or without (61.1%) the application of CS. Median follow-up was 4.5 years. Single enucleation showed a recurrence rate of 50%, but the additional application of CS reduced the recurrence rate to 14.3%. No detrimental effects of CS on the mandibular nerve were detected. Enucleation plus the application of CS reduced the recurrence rate of KCOTs compared with simple enucleation. The application of CS did not cause any damage to the mandibular nerve. PMID- 19294437 TI - Insights into the functional role of protonation states in the HIV-1 protease BEA369 complex: molecular dynamics simulations and free energy calculations. AB - The molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) method combined with molecular dynamics (MD) simulations were used to investigate the functional role of protonation in human immunodeficiency virus type 1 (HIV-1) protease complexed with the inhibitor BEA369. Our results demonstrate that protonation of two aspartic acids (Asp25/Asp25') has a strong influence on the dynamics behavior of the complex, the binding free energy of BEA369, and inhibitor-residue interactions. Relative binding free energies calculated using the MM-PBSA method show that protonation of Asp25 results in the strongest binding of BEA369 to HIV 1 protease. Inhibitor-residue interactions computed by the theory of free energy decomposition also indicate that protonation of Asp25 has the most favorable effect on binding of BEA369. In addition, hydrogen-bond analysis based on the trajectories of the MD simulations shows that protonation of Asp25 strongly influences the water-mediated link of a conserved water molecule, Wat301. We expect that the results of this study will contribute significantly to binding calculations for BEA369, and to the design of high affinity inhibitors. PMID- 19294438 TI - Findings in multi-detector row CT with portal phase enhancement in patients with mesenteric venous thrombosis. AB - There exists no systematic evaluation on the findings in multi-detector row computed tomography (MDCT) with portal phase enhancement in patients with mesenteric venous thrombosis (MVT). Patients with MVT between 2004 and 2006 were identified at Malmo University Hospital, Sweden. Thirty patients had MVT. Median age was 58 years (interquartile range, 46-72), and 57% (17) were men. MDCT with portal phase enhancement showed MVT in 19 out of 20 investigated patients at first evaluation, even though there was clinical suspicion in only one, and conservative management was possible in 19 (95%). Central and peripheral MVT was seen in 20 (100%) and six (30%) cases, respectively. Extra-hepatic and intra hepatic portal and splenic vein thrombosis was seen in 17 (85%), nine (45%) and 15 (75%) cases, respectively. Venous collaterals were visible in 12 (60%) cases. The most frequent extra-vascular pathologies were mesenteric oedema (50%), ascites (40%), small bowel wall oedema (25%) and local small bowel wall dilatation (20%). The vascular findings in MVT are more pronounced than the intestinal findings in MDCT with portal phase enhancement, and evaluation of the mesenteric vessels should be included in routine MDCT scans for unclear acute abdomen. PMID- 19294439 TI - A comparison of electrically evoked and channel rhodopsin-evoked postsynaptic potentials in the pharyngeal system of Caenorhabditis elegans. AB - Dissecting the function of neural circuits requires the capability to stimulate and record from the component neurones. Optimally, the methods employed should enable precise activation of distinct elements within the circuit and high fidelity readout of the neuronal response. Here we compare two methods for neural stimulation in the pharyngeal system of Caenorhabditis elegans by evoking postsynaptic potentials (PSPs) either by electrical stimulation or by expression of the channelrhodopsin [ChR2(gf)] in cholinergic neurones of the pharyngeal circuit. Using a dissection that isolates the pharynx and its embedded neural system of 20 neurones permits analysis of the neurotransmitter pathways within this microcircuit. We describe protocols for selective electrically evoked or ChR2-mediated cholinergic synaptic events in this circuit. The latter was achieved by generating strains, punc-17::ChR2(gf);yfp, that express ChR2(gf) in cholinergic neurones. PSPs evoked by both electrical and light stimulation exhibited a rapid time-course and were blocked by cholinergic receptor antagonists and rapidly reversed on cessation of the stimulus. Electrically evoked PSPs were also reduced in a hypomorphic mutant for the synaptic vesicle acetylcholine transporter, unc-17, further indicating they are nicotinic cholinergic PSPs. The pharyngeal nervous system is exquisitely sensitive to both electrical and light activation. For the latter, short light pulses of 200 mus delivered to punc-17::ChR2(gf);yfp are capable of generating full muscle action potentials. We conclude that the application of optogenetic approaches to the C. elegans isolated pharynx preparation opens the way for a precise molecular dissection of synaptic events in the pharyngeal microcircuit by providing a molecular and system level analysis of the synapses that control the feeding behaviour of C. elegans. PMID- 19294440 TI - Markers of inflammation collocate with increased wall stress in human coronary arterial plaque. AB - In this study, we hypothesized that spatial relationships exist between the local mechanical environment and inflammatory marker expression in atherosclerotic plaques, and that these relationships are plaque-progression dependent. Histologic cross-sections were collected at regular intervals along the length of diseased human coronary arteries and classified as early, intermediate, advanced, or mature based on their morphological features. For each cross-section, the spatial distribution of stress was determined using a 2D heterogeneous finite element model, and the corresponding distribution of selected inflammatory markers (macrophages, matrix metalloproteinase-1 [MMP-1], and nuclear factor kappa B [NF-kappaB]) were determined immunohistochemically. We found a monotonic spatial relationship between mechanical stress and activated NF-kappaB that was consistent in all stages of plaque progression. We also identified progression dependent relationships between stress and both macrophage presence and MMP-1 expression. These findings add to our understanding of the role of mechanical stress in stimulating the inflammatory response, and help explain how mechanical factors may regulate complex biological changes in remodeling. PMID- 19294441 TI - Production of bioethanol by direct bioconversion of oil-palm industrial effluent in a stirred-tank bioreactor. AB - The purpose of this study was to evaluate the feasibility of producing bioethanol from palm-oil mill effluent generated by the oil-palm industries through direct bioconversion process. The bioethanol production was carried out through the treatment of compatible mixed cultures such as Thrichoderma harzianum, Phanerochaete chrysosporium, Mucor hiemalis, and yeast, Saccharomyces cerevisiae. Simultaneous inoculation of T. harzianum and S. cerevisiae was found to be the mixed culture that yielded the highest ethanol production (4% v/v or 31.6 g/l). Statistical optimization was carried out to determine the operating conditions of the stirred-tank bioreactor for maximum bioethanol production by a two-level fractional factorial design with a single central point. The factors involved were oxygen saturation level (pO(2)%), temperature, and pH. A polynomial regression model was developed using the experimental data including the linear, quadratic, and interaction effects. Statistical analysis showed that the maximum ethanol production of 4.6% (v/v) or 36.3 g/l was achieved at a temperature of 32 degrees C, pH of 6, and pO(2) of 30%. The results of the model validation test under the developed optimum process conditions indicated that the maximum production was increased from 4.6% (v/v) to 6.5% (v/v) or 51.3 g/l with 89.1% chemical-oxygen-demand removal. PMID- 19294442 TI - Tuberculoma of the myocardium: a rare case of intra-vitam diagnosis. PMID- 19294443 TI - Chronic total coronary occlusions in patients with stable angina pectoris: impact on therapy and outcome in present day clinical practice. AB - BACKGROUND: Chronic total coronary occlusions (CTOs) represent a subgroup of coronary lesions with a low procedural success and high recurrence rate. However, there is evidence for a prognostic benefit of revascularizing a CTO. OBJECTIVE: This study assessed the prevalence of CTOs among patients with stable angina pectoris and its impact on therapeutic strategies. METHODS: Between 2001 and 2003, a survey was conducted in 64 sites to analyze the outcome of the first diagnostic angiography in patients presenting with stable angina pectoris (STAR registry). The clinical characteristics, initial angiographic findings, therapeutic strategy and outcome within the first year were analyzed. RESULTS: A total of 2,002 patients were entered into the registry. One-third had at least one CTO. At 1 year, the mortality in patients with a CTO was significantly higher than in those without a CTO (5.5 vs. 3.1%; P = 0.009). This excess mortality was related to a higher prevalence of confounding factors in patients with a CTO such as diabetes and more severe LV dysfunction. Patients with a CTO were more likely to undergo surgery or being treated medically, whereas patients without a CTO were more likely to undergo PCI. If a CTO was treated by PCI the periprocedural and long-term outcome was similar to those with PCI for a non-occlusive lesion. However, periprocedural MACE was higher for patients treated for a non-occlusive lesion without first treating the CTO. CONCLUSIONS: The prevalence of CTOs in patients with stable angina pectoris is high, and it influences the clinical outcome within the first year. The therapeutic strategy is influenced towards a rather conservative approach and lower rates of interventional therapy. PMID- 19294444 TI - Effects of candesartan cilexetil "add-on" treatment in congestive heart failure outpatients in daily practice. AB - AIMS: In the present study, we investigated the efficacy and safety of candesartan cilexetil (candesartan) as "add-on" treatment in congestive heart failure (CHF) in daily practice. METHODS AND RESULTS: In this open-label, multicenter study 414 CHF outpatients (NYHA II/III) with left ventricular ejection fraction (LVEF) < or = 40% and plasma brain natriuretic peptide (BNP) levels > 200 pg/ml at baseline were enrolled. Patients were treated with standard therapy including at least one angiotensin converting enzyme inhibitor in addition to another CHF drug; 91% of the patients received beta-blockers. Candesartan was uptitrated to 32 mg/day (target dose if tolerated) during 6 weeks followed by constant dosing over 16 weeks. The primary endpoint plasma BNP was significantly reduced by 25% at week 22 (from 394 to 295 pg/ml, P < 0.0001 vs. baseline). Candesartan produced early and sustained improvements of plasma BNP/NT pro-BNP, LVEF, and quality of life (SF-36) compared to baseline. Of patients on beta-blockers, 37% improved towards NYHA II/I at week 22 (P < 0.0001) and 53.5% of the patients in NYHA III at baseline improved into NYHA II/I at week 22 (n = 232, P < 0.0001). Candesartan was well tolerated; no unexpected findings were reported besides known adverse reactions including hypotension, hyperkalemia, and serum creatinine elevations. CONCLUSION: Candesartan "add-on" treatment provides a good benefit/risk ratio in CHF outpatients in daily practice, although high risk patients should be managed with frequent monitoring of BP, serum potassium, and renal function. PMID- 19294445 TI - Regulation of expression of apolipoprotein A-I by selenium status in human liver hepatoblastoma cells. AB - BACKGROUND: Cardiomyopathy is common to areas with low selenium (Se) intake and in patients receiving total parenteral nutrition. Although controversial, a few studies have suggested a protective role for Se in coronary heart disease on the basis of modulation of high-density lipoproteins (HDL). AIMS OF THE STUDY: In this study, the role of Se as a positive regulator of expression of a key HDL, apolipoprotein A-I (apoA-I), has been evaluated in human hepatoblastoma (HepG2) cell culture model. We further examined if the transcription of apoA-I, driven by the nuclear hormone receptor, peroxisome-proliferator activated receptor, PPARalpha, was trans-repressed by the presence of the oxidative stress-responsive transcription factor, NF-kappaB. METHODS: Modulation of expression of apoA-I and activation of nuclear NF-kappaB subunit p65 and PPARalpha by Se status were evaluated by Western blot and luciferase-based assays. Interaction of p65 with PPARalpha was evaluated by immunoprecipitation. RESULTS: HepG2 cultured in media with Se (100 nM) demonstrated an increase in the expression of apoA-I when compared to Se-deficient cells. A similar trend was also seen in mice that were supplemented with 0.4 ppm of Se as sodium selenite. Treatment of Se-supplemented cells with bacterial lipopolysaccharide (LPS) showed induction of apoA-I. Supplementation of hepatocytes with Se decreased the nuclear levels of p65, which prevented its interaction with PPARalpha to modulate apoA-I transcription. CONCLUSION: Our results suggest that supplementation of hepatocytes with Se mitigates oxidative stress-dependent repression of apoA-I expression by suppressing the NF-kappaB pathway, which allows PPARalpha to effectively drive the expression of apoA-I. PMID- 19294446 TI - The metabolic modulators, Etomoxir and NVP-LAB121, fail to reverse pressure overload induced heart failure in vivo. AB - Shifting substrate oxidation in heart muscle from fatty acids to glucose (substrate-switch) may improve contractile function in heart failure. We tested whether application of two agents (etomoxir and NVP-LAB121) capable of inducing a substrate-switch reverts the onset of heart failure in rats with chronic pressure overload. Hypertrophy was induced by aortic banding in rats for 1 or 15 weeks. Rats were treated for 10 days with the CPT-1-inhibitor etomoxir [29.5 micromol/(kg day)] or with NVP-LAB121 [60 micromol/(kg day)], a pyruvate dehydrogenase-kinase-inhibitor, before assessment by echocardiography and perfusion as isolated working hearts. We also analyzed PDH- and CPT1-activity and expression of alpha- and beta-MHC by RT-PCR. Aortic banding increased heart-to body-weight-ratio (g/kg) from 3.44 +/- 0.26 to 4.14 +/- 0.48 after 1 week and from 2.80 +/- 0.21 to 6.54 +/- 0.26 after 15 weeks. Ejection fraction was impaired after 15 weeks (57 +/- 11 vs. 73 +/- 8%, P < 0.05) and rats exhibited signs of heart failure. Total PDH activity was the same in all groups. CPT-1 activity was unchanged after 1 week but decreased after 15 weeks (P < 0.01). Neither etomoxir nor NVP-LAB121 affected cardiac function in vivo, but etomoxir improved function of the isolated heart. The drugs did not affect total PDH and CPT-1 activity, but increased PDH-activity status, prevented a decrease in PDK4 expression in heart failure, increased alpha and beta-MHC expression and shifted substrate oxidation toward glucose in the isolated working rat heart. In conclusion, pharmacologic induction of substrate-switching is associated with changes in myofibrillar isoform expression but does not reverse heart failure in vivo. The improvement of function in vitro deserves further investigation. PMID- 19294447 TI - Differences in hypothalamic-pituitary-adrenal axis functioning among children with ADHD predominantly inattentive and combined types. AB - Some evidence suggests that the HPA axis may be dysfunctional in children with attention-deficit/hyperactivity disorder (ADHD). The aim of this study was to investigate whether a different pattern of HPA axis activity is found between the inattentive (I) and combined (C) subtypes of ADHD, in comparison with healthy control children. A total of 100 prepubertal subjects [52 children with ADHD combined type (ADHD-C), 23 children with ADHD predominantly inattentive type (ADHD-I), and 25 healthy control subjects] were studied. The effects of stress were studied by comparing cortisol responses to a psychosocial stressor, consisting of a public speaking task. Children with ADHD-I showed an elevated cortisol response to the psychosocial stressor, in contrast to children with ADHD C who showed a blunted cortisol response to the psychosocial stressor. When a distinction was made between responders and non-responders (a subject was classified as a responder when there was an increase in cortisol reactivity), hyperactivity symptoms were clearly related to a lower cortisol reactivity to stress. The results indicate that a low-cortisol responsivity to stress may be a neurobiological marker for children with ADHD-C, but not for those with ADHD-I. Directions for future research and clinical implications are discussed. PMID- 19294448 TI - Surface aggregation of urinary proteins and aspartic Acid-rich peptides on the faces of calcium oxalate monohydrate investigated by in situ force microscopy. AB - The growth of calcium oxalate monohydrate in the presence of Tamm-Horsfall protein (THP), osteopontin, and the 27-residue synthetic peptides (DDDS)(6)DDD and (DDDG)(6)DDD (D = aspartic acid, S = serine, and G = glycine) was investigated via in situ atomic force microscopy. The results show that these four growth modulators create extensive deposits on the crystal faces. Depending on the modulator and crystal face, these deposits can occur as discrete aggregates, filamentary structures, or uniform coatings. These proteinaceous films can lead to either the inhibition of or an increase in the step speeds (with respect to the impurity-free system), depending on a range of factors that include peptide or protein concentration, supersaturation, and ionic strength. While THP and the linear peptides act, respectively, to exclusively increase and inhibit growth on the (101) face, both exhibit dual functionality on the (010) face, inhibiting growth at low supersaturation or high modulator concentration and accelerating growth at high supersaturation or low modulator concentration. Based on analyses of growth morphologies and dependencies of step speeds on supersaturation and protein or peptide concentration, we propose a picture of growth modulation that accounts for the observations in terms of the strength of binding to the surfaces and steps and the interplay of electrostatic and solvent induced forces at the crystal surface. PMID- 19294449 TI - Sequence- or position-specific mutations in the carboxyl-terminal FL motif of the kidney sodium bicarbonate cotransporter (NBC1) disrupt its basolateral targeting and alpha-helical structure. AB - The sodium-bicarbonate cotransporter NBC1 is targeted exclusively at the basolateral membrane. Mutagenesis of a dihydrophobic FL motif (residues 1013 1014) in the C-terminal domain disrupts the targeting of NBC1. In the present study, we determined the precise constraints of the FL motif required for basolateral targeting of NBC1 by expressing epitope-tagged wild-type and mutant NBC1 in MDCK cells and RNA-injected Xenopus oocytes and examining their subcellular localization. We assayed the functional activity of the mutants by measuring bicarbonate-induced currents in oocytes. Wild-type NBC1 (containing PFLS) was expressed exclusively on the basolateral membrane in MDCK cells. Reversal of the FL motif (PLFS) had no effect on basolateral targeting or activity. Shifting the FL motif one residue upstream (FLPS) resulted in mistargeting of the apical membrane but the FLPS mutant retained its functional activity in oocytes. Shifting the FL motif one residue downstream resulted in a mutant (PSFL) that did not efficiently translocate to the plasma membrane and was instead colocalized with the ER marker, protein disulfide isomerase (PDI). Analysis of circular dichroism (CD) revealed that a short peptide, 20 amino acid residues, of wild-type NBC1 contained a significant alpha-helical structure, whereas peptides in which the FL motif was reversed or C-terminally shifted were disordered. We therefore propose that the specific orientation and the precise location of the FL motif in the primary sequence of NBC1 are strict requirements for the alpha-helical structure of the C-terminal cytoplasmic domain and for targeting of NBC1 to the basolateral membrane. PMID- 19294450 TI - Modulation of erythrocyte acetylcholinesterase activity and its association with G protein-band 3 interactions. AB - Circulating acetylcholine, substrate of membrane acetylcholinesterase (AChE), is known to enhance the band 3 protein degree of phosphorylation. The purpose of this study was to verify whether the band 3 phosphorylation status is associated with a G protein and whether it is an influent factor on AChE enzyme activity. From blood samples of healthy donors, erythrocyte suspensions were prepared and incubated with AChE substrate (acetylcholine) and inhibitor (velnacrine), along with protein tyrosine kinase (PTK) and tyrosine phosphatase (PTP) inhibitors. AChE activity was determined by spectrophotometry and extract samples were analyzed by western blotting using primary antibodies to different G protein subunits. Our results with phosphorylated band 3 (PTP inhibitor) show an increase in erythrocyte AChE (p < 0.0001). A dephosphorylated band 3 state (PTK inhibitor) shows a significant decrease. We identified a potential linkage of protein subunits Galpha(i1/2) and G(beta) with band 3 protein. Galpha(i1/2) and G(beta) may be linked to the band 3 C-terminal site. Galpha(i1/2) is associated with the band 3 N-terminal domain, except for the control and ACh aliquots. G(beta) is associated with both phosphorylated and dephosphorylated band 3 in the presence of velnacrine. We conclude that an erythrocyte G protein with subunits Galpha(i1/2) and G(beta) is associated with band 3. AChE depends on the degree of band 3 phosphorylation and its association with Galpha(i1/2) and G(beta). PMID- 19294451 TI - Evidence for the transport of maltose by the sucrose permease, CscB, of Escherichia coli. AB - The purpose of this study was to examine the sugar recognition and transport properties of the sucrose permease (CscB), a secondary active transporter from Escherichia coli. We tested the hypothesis that maltose transport is conferred by the wild-type CscB transporter. Cells of E. coli HS4006 harboring pSP72/cscB were red on maltose MacConkey agar indicator plates. We were able to measure "downhill" maltose transport and establish definitive kinetic behavior for maltose entry in such cells. Maltose was an effective competitor of sucrose transport in cells with CscB, suggesting that the respective maltose and sucrose binding sites and translocation pathways through the CscB channel overlap. Accumulation ("uphill" transport) of maltose by cells with CscB was profound, demonstrating active transport of maltose by CscB. Sequencing of cscB encoded on plasmid pSP72/cscB used in cells for transport studies indicate an unaltered primary CscB structure, ruling out the possibility that mutation conferred maltose transport by CscB. We conclude that maltose is a bona fide substrate for the sucrose permease of E. coli. Thus, future studies of sugar binding, transport, and permease structure should consider maltose, as well as sucrose. PMID- 19294452 TI - Tentacles of venom: toxic protein convergence in the Kingdom Animalia. AB - The origin and evolution of venom in many animal orders remain controversial or almost entirely uninvestigated. Here we use cDNA studies of cephalopod posterior and anterior glands to reveal a single early origin of the associated secreted proteins. Protein types recovered were CAP (CRISP, Antigen 5 [Ag5] and Pathogenesis-related [PR-1]), chitinase, peptidase S1, PLA(2) (phospholipase A(2)), and six novel peptide types. CAP, chitinase, and PLA(2) were each recovered from a single species (Hapalochlaena maculosa, Octopus kaurna, and Sepia latimanus, respectively), while peptidase S1 transcripts were found in large numbers in all three posterior gland libraries. In addition, peptidase S1 transcripts were recovered from the anterior gland of H. maculata. We compare their molecular evolution to that of related proteins found in invertebrate and vertebrate venoms, revealing striking similarities in the types of proteins selected for toxic mutation and thus shedding light on what makes a protein amenable for use as a toxin. PMID- 19294453 TI - Tandem stop codons in ciliates that reassign stop codons. AB - Tandem stop codons are extra stop codons hypothesized to be present downstream of genes to act as a backup in case of read-through of the real stop codon. Although seemingly absent from Escherichia coli, recent studies have confirmed the presence of such codons in yeast. In this paper we will analyze the genomes of two ciliate species--Paramecium tetraurelia and Tetrahymena thermophila--that reassign the stop codons TAA and TAG to glutamine, for the presence of tandem stop codons. We show that there are more tandem stop codons downstream of both Paramecium and Tetrahymena genes than expected by chance given the base composition of the downstream regions. This excess of tandem stop codons is larger in Tetrahymena and Paramecium than in yeast. We propose that this might be caused by a higher frequency of stop codon read-through in these species than in yeast, possibly because of a leaky termination machinery resulting from stop codon reassignment. PMID- 19294455 TI - Growth and elemental accumulation of plants grown in acidic soil amended with coal fly ash-sewage sludge co-compost. AB - A greenhouse experiment was conducted to evaluate the growth and heavy-metal accumulation of Brassica chinensis and Agropyron elongatum in 10 and 25% ash sludge co-compost (ASC)--amended loamy acidic soil (pH 4.51) at two different application rates: 20% and 40% (v/v). Soil pH increased, whereas electrical conductivity decreased with the amendment of ASC to soil. Bioavailable Cu, Zn, and Mn contents of ASC-amended soil decreased, whereas Ni, Pb, and B contents increased. Concentrations of bioavailable Cu, Zn, and Mn in sludge compost (SC) amended soils were 5.57, 20.8, and 8.19 mg kg(-1), respectively. These concentrations were significantly lower than those in soil receiving an application rate of 20 or 25% ASC as 2.64, 8.48, and 5.26 mg kg(-1), respectively. Heavy metals and B contents of the composting mass significantly increased with an increase in ASC application rate from 20 to 40% (6.2 to 16.6 mg kg(-1) for 10% ASC- and 9.4 to 18.6 mg kg(-1) for 25% ASC-amended soil. However, when the ash content in co-compost increased from 10 to 25% during composting, bioavailable heavy-metal contents decreased. However, B contents increased with an increase in ash content. Addition of co-composts increased the dry-weight yield of the plants, and this increase was more obvious as the ash amendment rate in the co-composts and the ASC application rate increased. In case of B. chinensis, the biomass of 2.84 g/plant for 40% application of 25% ASC was significantly higher than SC (0.352 g/plant), which was 40% application of 10% ASC (0.434 g/plant) treatments. However, in A. elongatum, the differences between biomass of plants grown with 10% (1.34-1.94 g/ plant) and 25% ASC (2.12-2.21 g/plant) were not significantly different. Furthermore, there were fewer plant available heavy metals in 25% ASC, which decreased the uptake of heavy metals by plants. ASC was favorable in increasing the growth of B. chinensis and A. elongatum. The optimal ash amendment to the sludge composting and ASC application rates were at 25 and 20%, respectively. PMID- 19294454 TI - Evolution of the mitochondrial genome in mammals living at high altitude: new insights from a study of the tribe Caprini (Bovidae, Antilopinae). AB - Organisms living at high altitude are exposed to severe environmental stress associated with decreased oxygen pressure, cold temperatures, increased levels of UV radiation, steep slopes, and scarce food supplies, which may have imposed important selective constraints on the evolution of the mitochondrial genome. Within mammals, the tribe Caprini is of particular interest for studying the evolutionary effects of life at high altitude, as most species live in mountain regions, where they developed morphological and physiological adaptations for climbing. In this report, we analyzed the complete mitochondrial genome of 24 ruminants, including 20 species of Caprini. The phylogenetic analyses based on 16,117 nucleotides suggested the existence of a new large clade, here named subtribe Caprina, containing all genera, but Pantholops (Pantholopina), Capricornis, Naemorhedus, and Ovibos (Ovibovina). The alignment of the control region showed that all Caprini have between two and four tandem repeats of ~75 bp in the RS2 region, and that several of these copies emerged from recent and independent duplication events. We proposed therefore that the maintenance of at least two RS2 repeats in the control region of Caprini is positively selected, probably for producing a higher number of D-loop strands 3'-ending at different locations. The analyses of base composition at third-codon positions of protein coding genes revealed that Caprini have the highest percentage of A nucleotide and the lowest percentage of G nucleotide, a pattern which suggests increased rates of cytosine deamination (C-->T transitions) on the H strand of mtDNA. Two nonexclusive hypotheses related to high-altitude life can explain such a mutational pattern: more severe oxidative stress (ROS) and higher metabolic rates. By comparing the relative rates of nonsynonymous and synonymous substitutions in protein-coding genes, we identified that Caprini have higher levels of adaptive variation in the ATPase complex. In addition, we detected several changes in mitochondrial genes that should be tested for their potential role in mountain adaptation. PMID- 19294456 TI - Antioxidant enzyme activity in bacterial resistance to nicotine toxicity by reactive oxygen species. AB - We analyzed superoxide dismutase (SOD), catalase (CAT), and ATPase activities in the highly nicotine-degrading strain Pseudomonas sp. HF-1 and two standard strains Escherichia coli and Bacillus subtilis in an attempt to understand antioxidant enzymes in bacteria are produced in response to nicotine, which increases the virulence of the bacteria. Nicotine had different effects on different antioxidant enzymes of different bacteria. SOD plays a more important role in resistance to nicotine stress in E. coli than it does in CAT. Multiple antioxidant enzymes are involved in combating oxidative stress caused by nicotine in Pseudomonas sp. HF-1. The contribution of a particular antioxidant enzyme for protection from nicotine stress varies with the growth phase involved. The inhibition of ATPase in Pseudomonas sp. HF-1 at the stationary phase was enhanced with increasing nicotine concentration, showing a striking dose-response relationship. Nicotine probably affected the metabolism of ATP to some extent. Furthermore, different bacteria possessed distinct SOD isoforms to cope with oxidative stress caused by nicotine. PMID- 19294457 TI - Arterial tortuosity syndrome. PMID- 19294458 TI - Serpigenous calcification in the heart. PMID- 19294459 TI - Carotid artery stent migration in a child with Williams syndrome. AB - We report a case of distal migration of a stent from the brachiocephalic artery to the distal right common carotid artery 7 months after implantation in a 5-year old child with Williams syndrome. There were no neurological sequelae and the migrated stent remained widely patent 5 years following implantation. PMID- 19294460 TI - Xihong Hu, Guoying Huang, Mier Pa. Multidetector computed tomography for assessing a pulmonary artery sling in a pediatric patient. Pediatr Cardiol 2008; 29:1006-1007. PMID- 19294461 TI - Atrial tachyarrhythmias in neonatal enterovirus myocarditis. AB - Enterovirus myocarditis can be a serious infection in neonates, complicated by cardiac dysfunction and arrhythmias. Although decreased systolic function and ventricular arrhythmias have been reported, there is limited description of significant atrial involvement. We present an unusual series of three infants at our institution with enterovirus myocarditis in the neonatal period associated with atrial arrhythmias and dilation. PMID- 19294462 TI - Successful transcatheter closure of a large patent ductus venosus with the Amplatzer vascular plug II. AB - Patent ductus venosus is a rare form of congenital portosystemic shunt from the fetal umbilical vein to the inferior vena cava. The reported surgical treatments include ligation, banding, and liver transplantation. In addition, transcatheter closure with a coil, stent, or original Amplatzer vascular plug (AVP) has been reported. The AVP II, a redesigned version of the original vascular plug with a finer more densely woven nitinol wire and a large diameter (up to 22 mm) is available. This reported case is the first successful occlusion of a large patent ductus venosus with the new AVP II. PMID- 19294463 TI - Killing of Gram-negative bacteria with normal human serum and normal bovine serum: use of lysozyme and complement proteins in the death of Salmonella strains O48. AB - Serum is an environment in which bacterial cells should not exist. The serum complement system provides innate defense against microbial infections. It consists of at least 35 proteins, mostly in pre-activated enzymatic forms. The activation of complement is achieved through three major pathways: the classical, alternative, and lectin. Lysozyme, widely present in body fluids, catalyzes the hydrolysis of beta 1,4 linkage between N-acetyloglucosamine and N-acetylmuramic acid in the bacterial cell wall and cooperates with the complement system in the bactericidal action of serum. In this study, ten strains of serotype O48 Salmonella, mainly associated with warm-blooded vertebrates and clinically important causing diarrhea in infants and children, were tested. The results demonstrated that the most efficient killing of Salmonella O48 occurred when all the components of normal bovine serum (NBS) and normal human serum (NHS) cooperated. To prove the role of lysozyme in the bactericidal activity of bovine and human serum, the method of serum adsorption onto bentonite (montmorillonite, MMT) was used. In order to investigate structural transitions accompanying the adsorption of serum components, we applied X-ray diffraction methods. The results of this investigation suggested that apart from lysozyme, other proteins (as, e.g., C3 protein or IgG immunoglobulin) were adsorbed on MMT particles. It was also shown that Ca(2+) cations can be adsorbed on bentonite. This may explain the different sensitivities of the serovars belonging to the same O48 Salmonella serotype to NBS and NHS devoid of lysozyme. PMID- 19294464 TI - Intrabile duct metastasis from colonic adenocarcinoma without liver parenchyma involvement: contrast enhanced ultrasonography detection. AB - It is well-known that biliary duct invasion with intraluminal growth is one of the developmental patterns of primary liver tumors, and macroscopic intrabiliary growth of liver metastases in colorectal cancer is found with high frequency. Surgical treatment is the only potential curative therapy. However, many patients die of intrahepatic and/or extrahepatic recurrence after the resection. One of the causes of high recurrence rate after resective surgery, particularly surgical margin recurrences, is the invasion of biliary ducts mainly due to intraluminal tumor growth. We describe the first recorded case of a metastasis from colorectal cancer involving solely the common hepatic biliary duct, without invasion of contiguous liver parenchyma. A correct diagnosis was obtained by means of contrast enhanced ultrasound and ultrasound-guided fine needle aspiration biopsy. PMID- 19294465 TI - The Lyon-Turin high-speed rail: the public debate and perception of environmental risk in Susa Valley, Italy. AB - When the construction of the Lyon-Turin segment of the new European high-speed rail network was first publicly announced at the beginning of the 1990 s, it immediately found fierce opposition from the inhabitants of Susa Valley, Italy, one of the areas to be cut across by such infrastructure. At issue were the project's potential environmental impact and its consequences on public health. This study intends to clarify environmental risk perception and public debate between the national government, local advocacy groups, and the inhabitants of Susa Valley. Two major phases of public reaction were identified: (1) an initial rebellious period of no real dialog among the project's major stakeholders (exemplified by the popular "No TAV" [No High Speed Train] movement), followed by (2) a yielding period of intense multilateral negotiations centered on the activities of the "Lyon-Turin Environmental Observatory." The results of a qualitative cross analysis of the residents' perception of the proposed high speed rail revealed that public acceptance of risk in Susa Valley was influenced by the characteristics of hazards perceived by the residents and by the communicative approach used by the project's various stakeholders. It also emerged that early dialog among all the parties involved was critical in forming a personal viewpoint on risk, which, once consolidated, defied new information and perspectives. Likely, a greater and earlier care taken by the other stakeholders to inform and consult the local population about the railway would have greatly eased the public debate. PMID- 19294466 TI - Thymic neuroendocrine tumors in multiple endocrine neoplasia type 1: a comparative study on 21 cases among a series of 761 MEN1 from the GTE (Groupe des Tumeurs Endocrines). AB - BACKGROUND: Thymic neuroendocrine tumors (Th-NET) present a poor prognosis for patients with multiple endocrine neoplasia type 1 (MEN1). The purpose of this article was to study the clinical, biological, and pathological features of Th NET in a large cohort of patients with MEN1. METHODS: The 761-patient MEN1 cohort from the GTE registry was used (Groupe des Tumeurs Endocrines). RESULTS: The actuarial probability of occurrence was 2.6% (range, 1.3-5.5%) at aged 40 years. All, except one, Th-NET patients were men. Four patients had no other associated lesions. The youngest patient was aged 16 years. Mean age at the time of diagnosis was 42.7 (range, 16.1-67.5) years. The 10-year probability of survival was 36.1% (range, 11.5-62%). Seven patients (33%) belonged to clustered MEN1 families. The spectrum of associated lesions in patients with Th-NET was not statistically different from the spectrum of the remainder of the cohort. Various endocrine markers were high, but none were sensitive or specific enough to be useful for Th-NET detection. CT-scan and MRI were always positive at the time of diagnosis. No particular mutation was found to be associated with Th-NET. Five cases underwent prophylactic thymectomy without success. CONCLUSIONS: Several end points may be helpful for future guidelines: (1) earlier detection of Th-NET in MEN1 patients is required; (2) screening of both sexes is necessary; (3) a prospective study comparing MRI vs. CT scan in yearly screening for Th-NET is needed; (4) a reinforced screening program must be established for patients who belong to clustered families; and (5) thymectomies must be performed in specialized centers. PMID- 19294467 TI - Intrahepatic cholangiocarcinoma: prognostic factors after surgical resection. AB - BACKGROUND: Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver tumor. The resectability rate is low because at the time of diagnosis this disease is frequently beyond the limits of surgical therapy. Curative resection (R0) is the most effective treatment and the only therapy associated with prolonged disease-free survival. Based on the gross appearance of the tumor the Liver Cancer Study Group of Japan (LCSGJ) defined three types: mass forming type (MF), periductal infiltrating type (PI), intraductal growth (IG) type. The prognostic significance of gross type has been demonstrated in Eastern countries, but this issue has not been clarified in Western countries. The aim of this study was to identify the prognostic factors for survival in a group of patients submitted to surgical resection for ICC. METHODS: Between 1990 and 2007 a total of 81 consecutive patients with ICC were submitted to surgery. Patients with peritoneal carcinomatosis, extensive vascular involvement, or multiple intrahepatic metastases were excluded from surgical resection. Tumors were classified according to TMN stage (6th edition, 2002) and LCSGJ gross type classification. Tumor gross appearance on the cut surface was categorized into the following types according to the classification proposed by the Liver Cancer Study Group of Japan: MF, PI, or IG type. RESULTS: During the study period 52 patients were submitted to surgical resection with curative intent, whereas in 29 patients surgery was limited to explorative laparotomy. Curative resection (R0) was achieved in 43 patients (83%); and a major hepatic resection was performed in 63% (33/52) of the patients. Extrahepatic bile duct resection was carried out in 36% (19/52) of cases. According to the LCSGJ classification, the MF type was present in 34 patients (65%), the MF + PI type in 13 (25%), the PI type in 3 (6%), and the IG type in 2 (4%). Overall median survival time was 40 months, with a 1-, 3-, and 5-year actuarial survival rates of 83%, 50%, 20%, respectively. Survival was significantly related to the macroscopic gross type, with a median survival of 50 months for patients with the MF type, 19 months for the MF + PI type, 15 months for the PI type, and 17 months for the IG type. At univariate analysis, the macroscopic gross appearance of the tumor, the presence of lymph node metastasis, involvement of extrahepatic bile ducts, the presence of macroscopic vascular invasion, and positive resection margins were significant related to survival. At multivariate analysis, macroscopic vascular invasion and lymph nodes metastases were significant related to survival with hazard ratios of 4.11 and 2.79, respectively. Further statistical analyses were carried out to identify the relation between macroscopic gross type and prognosis. We identified that the MF + PI type tumors were significantly associated with negative prognostic factors, such as the involvement of extrahepatic bile ducts, the presence of lymph nodes metastases, the presence of macroscopic vascular invasion, the presence of perineural invasion, and higher T stage. CONCLUSIONS: Curative resection of ICC is the only therapy that can achieve long-term survival. The best results were observed in patients who underwent R0 resection for MF tumors without lymph node metastases or vascular invasion. Important predictive factors related to poor survival are MF + PI macroscopic tumor type, lymph node metastases, and vascular invasion. In these patients, other therapeutic approaches (i.e., adjuvant or neoadjuvant therapy) should be evaluated to improve results. PMID- 19294468 TI - The caBIG annotation and image Markup project. AB - Image annotation and markup are at the core of medical interpretation in both the clinical and the research setting. Digital medical images are managed with the DICOM standard format. While DICOM contains a large amount of meta-data about whom, where, and how the image was acquired, DICOM says little about the content or meaning of the pixel data. An image annotation is the explanatory or descriptive information about the pixel data of an image that is generated by a human or machine observer. An image markup is the graphical symbols placed over the image to depict an annotation. While DICOM is the standard for medical image acquisition, manipulation, transmission, storage, and display, there are no standards for image annotation and markup. Many systems expect annotation to be reported verbally, while markups are stored in graphical overlays or proprietary formats. This makes it difficult to extract and compute with both of them. The goal of the Annotation and Image Markup (AIM) project is to develop a mechanism, for modeling, capturing, and serializing image annotation and markup data that can be adopted as a standard by the medical imaging community. The AIM project produces both human- and machine-readable artifacts. This paper describes the AIM information model, schemas, software libraries, and tools so as to prepare researchers and developers for their use of AIM. PMID- 19294470 TI - Ethanol and (-)-alpha-Pinene: attractant kairomones for bark and ambrosia beetles in the southeastern US. AB - In 2002-2004, we examined the flight responses of 49 species of native and exotic bark and ambrosia beetles (Coleoptera: Scolytidae and Platypodidae) to traps baited with ethanol and/or (-)-alpha-pinene in the southeastern US. Eight field trials were conducted in mature pine stands in Alabama, Florida, Georgia, North Carolina, and South Carolina. Funnel traps baited with ethanol lures (release rate, about 0.6 g/day at 25-28 degrees C) were attractive to ten species of ambrosia beetles (Ambrosiodmus tachygraphus, Anisandrus sayi, Dryoxylon onoharaensum, Monarthrum mali, Xyleborinus saxesenii, Xyleborus affinis, Xyleborus ferrugineus, Xylosandrus compactus, Xylosandrus crassiusculus, and Xylosandrus germanus) and two species of bark beetles (Cryptocarenus heveae and Hypothenemus sp.). Traps baited with (-)-alpha-pinene lures (release rate, 2-6 g/day at 25-28 degrees C) were attractive to five bark beetle species (Dendroctonus terebrans, Hylastes porculus, Hylastes salebrosus, Hylastes tenuis, and Ips grandicollis) and one platypodid ambrosia beetle species (Myoplatypus flavicornis). Ethanol enhanced responses of some species (Xyleborus pubescens, H. porculus, H. salebrosus, H. tenuis, and Pityophthorus cariniceps) to traps baited with (-)-alpha-pinene in some locations. (-)-alpha-Pinene interrupted the response of some ambrosia beetle species to traps baited with ethanol, but only the response of D. onoharaensum was interrupted consistently at most locations. Of 23 species of ambrosia beetles captured in our field trials, nine were exotic and accounted for 70-97% of total catches of ambrosia beetles. Our results provide support for the continued use of separate traps baited with ethanol alone and ethanol with (-)-alpha-pinene to detect and monitor common bark and ambrosia beetles from the southeastern region of the US. PMID- 19294469 TI - Integrated mercury monitoring program for temperate estuarine and marine ecosystems on the North American Atlantic coast. AB - During the past century, anthropogenic activities have altered the distribution of mercury (Hg) on the earth's surface. The impacts of such alterations to the natural cycle of Hg can be minimized through coordinated management, policy decisions, and legislative regulations. An ability to quantitatively measure environmental Hg loadings and spatiotemporal trends of their fate in the environment is critical for science-based decision making. Here, we outline a Hg monitoring program for temperate estuarine and marine ecosystems on the Atlantic Coast of North America. This framework follows a similar, previously developed plan for freshwater and terrestrial ecosystems in the U.S. Methylmercury (MeHg) is the toxicologically relevant form of Hg, and its ability to bioaccumulate in organisms and biomagnify in food webs depends on numerous biological and physicochemical factors that affect its production, transport, and fate. Therefore, multiple indicators are needed to fully characterize potential changes of Hg loadings in the environment and MeHg bioaccumulation through the different marine food webs. In addition to a description of how to monitor environmental Hg loads for air, sediment, and water, we outline a species-specific matrix of biotic indicators that include shellfish and other invertebrates, fish, birds and mammals. Such a Hg monitoring template is applicable to coastal areas across the Northern Hemisphere and is transferable to arctic and tropical marine ecosystems. We believe that a comprehensive approach provides an ability to best detect spatiotemporal Hg trends for both human and ecological health, and concurrently identify food webs and species at greatest risk to MeHg toxicity. PMID- 19294471 TI - High quality care and ethical pay-for-performance: a Society of General Internal Medicine policy analysis. AB - BACKGROUND: Pay-for-performance is proliferating, yet its impact on key stakeholders remains uncertain. OBJECTIVE: The Society of General Internal Medicine systematically evaluated ethical issues raised by performance-based physician compensation. RESULTS: We conclude that current arrangements are based on fundamentally acceptable ethical principles, but are guided by an incomplete understanding of health-care quality. Furthermore, their implementation without evidence of safety and efficacy is ethically precarious because of potential risks to stakeholders, especially vulnerable patients. CONCLUSION: We propose four major strategies to transition from risky pay-for-performance systems to ethical performance-based physician compensation and high quality care. These include implementing safeguards within current pay-for-performance systems, reaching consensus regarding the obligations of key stakeholders in improving health-care quality, developing valid and comprehensive measures of health-care quality, and utilizing a cautious evaluative approach in creating the next generation of compensation systems that reward genuine quality. PMID- 19294472 TI - A prospective controlled trial of the influence of a geriatrics home visit program on medical student knowledge, skills, and attitudes towards care of the elderly. AB - PURPOSE: To determine the impact of a geriatrics home visit program for third year medical students on attitudes, skills, and knowledge. METHODS: Using a mixed methods, prospective, controlled trial, volunteer control group students (n = 17) at two sites and intervention group students (n = 16) at two different sites within the same internal medicine clerkship were given Internet and CDROM-based geriatric self-study materials. Intervention group students identified a geriatrics patient from their clinical experience, performed one "home" visit (home, nursing home, or rehabilitation facility) to practice geriatric assessment skills, wrote a structured, reflective paper, and presented their findings in small-group teaching settings. Papers were qualitatively analyzed using the constant comparative method for themes. All students took a pre-test and post test to measure changes in geriatrics knowledge and attitudes. RESULTS: General attitudes towards caring for the elderly improved more in the intervention group than in the control group (9.8 vs 0.5%; p = 0.04, effect size 0.78). Medical student attitudes towards their home care training in medical school (21.7 vs 3.2%; p = 0.02, effect size 0.94) improved, as did attitudes towards time and reimbursement issues surrounding home visits (10.1 vs -0.2%; p = 0.02, effect size 0.89). Knowledge of geriatrics improved in both groups (13.4 vs 15.2% improvement; p = 0.73). Students described performing a mean of seven separate geriatric assessments (range 4-13) during the home visit. Themes that emerged from the qualitative analysis of the reflective papers added depth and understanding to the quantitative data and supported results concerning attitudinal change. CONCLUSIONS: While all participants gained geriatrics knowledge during their internal medicine clerkship, students who performed a home visit had improved attitudes towards the elderly and described performing geriatric assessment skills. Requiring little faculty time, a geriatrics home visit program like this one may be a useful clerkship addition to foster medical students' professional growth. PMID- 19294473 TI - SSAT controversies intramucosal esophageal cancer and high-grade dysplasia: which treatment? Surgical therapy: improved outcomes and piece of mind. PMID- 19294474 TI - Primary hepatic osteosarcoma. PMID- 19294475 TI - Migration of the connecting tube into small bowel after adjustable gastric banding. AB - Migration of intraabdominal catheters into the bowel is a well-known phenomenon of uncertain cause. Intragastric migration of adjustable gastric banding is not an uncommon complication, with a reported prevalence of 1-11% in most series. Migration of the connecting tube into the small bowel lumen or colonic lumen is very rare. We present a case study of a female who had adjustable gastric banding with persistent wound infection at the access port side that failed with conservative treatment. Migration of the connecting tube into the jejunal lumen was observed at laparoscopy. The connecting tube and port were removed and the band remained. Small bowel defects were repaired laparoscopically. Migration of band tubing into the bowel should be considered in the clinical setting of relapsing port-side infection. PMID- 19294476 TI - Proximal femoral reconstructions with bone impaction grafting and metal mesh. AB - Extensive circumferential proximal cortical bone loss is considered by some a contraindication for impaction bone grafting in the femur. We asked whether reconstruction with a circumferential metal mesh, impacted bone allografts, and a cemented stem would lead to acceptable survival in these patients. We retrospectively reviewed 14 patients (15 hips) with severe proximal femoral bone defects (average, 12 cm long; 14 type IV and one type IIIB using the classification of Della Valle and Paprosky) reconstructed with this method. The minimum followup was 20 months (average, 43.2 months; range, 20-72 months). Preoperative Merle D'Aubigne and Postel score averaged 4.8 points. With revision of the stem as the end point, the survivorship of the implant was 100% at one year and 86.6% at 72 months. The mean functional score at last followup was 14.4 points. We observed two fractures of the metal mesh at 31 and 48 months in cases reconstructed with a stem that did not bypass the mesh. Dislocation (3 cases) and acute deep infection (3 cases) were the most frequent complications. Patients with complete absence of the proximal femur may be candidates for biological proximal femoral reconstructions using this salvage procedure. Bone impaction grafting must be a routine technique if this method is selected. PMID- 19294477 TI - Amyopathic dermatomyositis or dermatomyositis-like skin disease: retrospective review of 16 cases with amyopathic dermatomyositis. AB - Amyopathic dermatomyositis (ADM) is characterized by the presence of dermatomyositis (DM) for 6 months or more in individuals who have normal muscle enzymes and no clinically significant muscle weakness. The aim of the study was to investigate the initial laboratory data, clinical manifestations, complications, and clinical outcomes of patients with the diagnosis of ADM. We reported 16 cases with the cutaneous findings of dermatomyositis without clinical or laboratory evidence of muscle disease for at least 2 years after onset of the skin manifestations in the Department of Dermatology and Rheumatology at Shanghai Ruijin Hospital between 1998 and 2004. All patients had Gottron's papules, periungual erythema/telangiectasia, and violaceous discoloration of the face, neck, upper chest, and back at some time during the course of their disease. Follow-up of 1 to 10 years after diagnosis found muscle weakness in three patients (18.75%) within 5 years of diagnosis. One patient (6.15%) was rediagnosed as chronic cutaneous lupus erythematosus (CCLE). Four patients (25%) had associated malignancies. Twelve patients (75%) had radiographic evidence indicative of interstitial fibrosis irrespective of respiratory symptoms. Patients with ADM appear to be at risk for developing the same potentially fatal disease complications as those patients with DM (e.g., interstitial lung disease and internal malignancy). These cases further emphasize that the cutaneous manifestations of dermatomyositis are pathognomonic for DM and we propose the term dermatomyositis-like skin disease as a better designation than amyopathic dermatomyositis to describe this distinctive subset of cutaneous symptoms. Dermatomyositis-like skin disease is a complex syndrome, which includes the characteristic cutaneous eruption of dermatomyositis without clinical evidence of muscle disease. Our findings suggest that patients diagnosed with this syndrome are at risk for fatal interstitial lung disease, malignancy, and/or delayed onset of DM or CCLE. Cautious systematic clinical trials should be considered for this group of patients. PMID- 19294478 TI - Discovertebral (Andersson) lesions of the spine in ankylosing spondylitis revisited. AB - A well-known complication in patients with ankylosing spondylitis (AS) is the development of localised vertebral or discovertebral lesions of the spine, which was first described by Andersson in 1937. Since then, many different terms are used in literature to refer to these localised lesions of the spine, including the eponym 'Andersson lesion' (AL). The use of different terms reflects an ongoing debate on the exact aetiology of the AL. In the current study, we performed an extensive review of the literature in order to align communication on aetiology, diagnosis and management between treating physicians. AL may result from inflammation or (stress-) fractures of the complete ankylosed spine. There is no evidence for an infectious origin. Regardless of the exact aetiology, a final common pathway exists, in which mechanical stresses prevent the lesion from fusion and provoke the development of pseudarthrosis. The diagnosis of AL is established on conventional radiography, but computed tomography and magnetic resonance imaging both provide additional information. There is no indication for a diagnostic biopsy. Surgical instrumentation and fusion is considered the principle management in symptomatic AL that fails to resolve from a conservative treatment. We advise to use the term Andersson lesion for these spinal lesions in patients with AS. PMID- 19294479 TI - Effects of a hand-joint protection programme with an addition of splinting and exercise: one year follow-up. AB - Forty-two women with thumb base osteoarthritis referred to a joint protection programme (JP) were distributed into groups: one with only JP (Control group) and one with addition of splints day/night, hot pack/home exercise (SE group). Assessments of pain, stiffness, grip force, disabilities of daily activities were performed before treatment, 1 week and 1 year after treatment. The SE group had a significant decrease in pain, stiffness and an improvement in daily activities directly after the intervention and at 1-year follow-up compared to the Control group. In the SE group pain at night, pain on motion, and stiffness decreased. Grip force increased and daily activities improved. The Control group decreased in pain on motion and showed improvement in daily activities just after the intervention but not at 1-year follow up. This comparative study shows that when splinting and exercise regimen are added to a JP programme it gives a greater improvement of pain, stiffness, grip force and daily activities than the JP programme alone. PMID- 19294480 TI - An unusual case of cancer of the urachal remnant following repair of bladder exstrophy. AB - INTRODUCTION: We report the first case of cancer of the urachal remnant following repair of bladder exstrophy, in a renal transplant recipient. METHOD: A retrospective review of this clinical case and the associated literature were performed. CONCLUSION: This unusual case highlights two very rare entities. Bladder exstrophy has an incidence of 1 in 50,000 newborns, whereas urachal cancer accounts for less than 1% of all bladder tumours. PMID- 19294481 TI - Student-selected components in surgery: providing practical experience and increasing student confidence. AB - INTRODUCTION: Reviews of the medical school curriculum in the UK and Ireland have recommended the introduction of student-selected components (SSCs). The Department of Surgery in The Royal College of Surgeons in Ireland (RCSI) has introduced a 6-week surgical SSC, which aims to develop practical clinical skills, provide mentorship and prepare students for internship. METHODS: Emphasis was placed on exposure to surgical specialities, teaching practical skills and student integration into surgical teams. Students completed an online survey pre- and post-SSC, assessing attitudes towards the course and confidence in performing ward-based and surgical skills. RESULTS: The mean Likert scale scores increased for all the skills assessed. Students felt that the SSC prepared them for their first day of work and strengthened their desire to pursue surgical careers. CONCLUSION: A surgical SSC has been successful in increasing student confidence in performing practical skills required for commencing work as a doctor. Provision of dedicated SSCs is likely to influence the career choice of students. PMID- 19294482 TI - Post-traumatic headache: facts and doubts. AB - The International Classification of Headache Disorders does not separate the moderate from severe/very severe traumatic brain injury (TBI), since they are all defined by Glasgow coma scale (GCS) < 13. The distinction between the severe and very severe TBI (GCS < 8) should be made upon coma duration that in the latter may be longer than 15 days up to months in the case of vegetative state. Post traumatic amnesia duration may double the coma duration itself. Therefore, the 3 month parameter proposed to define the occurrence or resolution of post-traumatic headache (PTH) appears inadequate. Following TBI, neuropathic pain, central pain, thalamic pain, combined pain are all possible and they call for proper pharmacological approaches. One more reason for having difficulties in obtaining information about headache in the early phase after regaining consciousness is the presence of concomitant medications that may affect pain perception. Post traumatic stress disorder (PTSD) develops days or weeks after stress and tends to improve or disappear within 3 months after exposure; interestingly, this spontaneous timing resembles that of PTH. In our experience the number of TBI patients with PTH at 1-year follow-up is lower in those with longer coma duration and more severe TBI. Cognitive functioning evaluated after at least 12 months from TBI, showed mild or no impairment in these patients with severe TBI and PTH, whereas they have psychopathological changes, namely anxiety and depression. The majority of patients with PTH after severe/very severe TBI had skull fractures or dural lacerations and paroxystic EEG abnormalities. The combination of psychological changes (depression and anxiety) and organic features (skull fractures, dural lacerations, epileptic EEG abnormalities) in PTH may be inversely correlated with the severity of TBI, with prevalence of psychological disturbances in mild TBI and of organic lesions in severe TBI. On the other hand, only in severe TBI patients with good cognitive recovery the influence of the psychopathological disorders may play a role. In fact, the affective pain perception is probably related to the integrity of cognitive functions as in mild TBI and in severe TBI with good cognitive outcome. PMID- 19294483 TI - The botanical origin of kratom (Mitragyna speciosa; Rubiaceae) available as abused drugs in the Japanese markets. AB - Kratom is the leaves of Mitragyna speciosa (Rubiaceae). Recently, kratom has been sold in street shops or on the Internet in Japan for the purpose of abuse due to its opium-like effects. In this study, we investigated the botanical origin of the commercial kratom products using the internal transcribed spacer (ITS) sequence analysis of rDNA in preparation for future regulation of this product. In addition, a previously reported method to authenticate the plant, utilizing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to the same products in order to estimate the method's accuracy and utility. The ITS sequence analysis of the commercial kratoms revealed that most of them were derived from M. speciosa or closely related plants, while the others were made from the same tribe plant as M. speciosa. The reported PCR-RFLP method could clearly distinguish kratoms from the other psychoactive plants available in the Japanese markets and also from related plants. The authentication method is considered to be useful for the practical regulation of the plant due to its wide range of application, high accuracy and simplicity. PMID- 19294484 TI - Rapid response and sustained remission by rituximab in four cases of plasma exchange-failed acute thrombotic thrombocytopenic purpura. PMID- 19294485 TI - T cell lymphoblastic lymphoma in parotidectomy for Warthin's tumor: case report and review of the literature. AB - Lymphomas associated with Warthin's tumor (WT) are extremely uncommon and the majorities are of B cell type. We report the simultaneous occurrence of T-cell lymphoblastic lymphoma (T-LBL) and WT in an 81-year-old patient, who presented with fever, night sweats and enlargement of the right parotid gland. The parotidectomy specimen showed a WT with extensive replacement of the lymphoid stroma by T-LBL, but preservation of the oncocytic epithelium. Staging investigations revealed mediastinal and abdominal lymphadenopathy, bilateral pleural effusions and bone marrow infiltration, in keeping with stage IVB disease. The patient received combination chemotherapy treatment but responded poorly, and died three months after diagnosis. To our knowledge, this is the first case report of T-LBL involving WT. The present study indicates that the lymphoid stroma in WT belongs to the systemic lymphoid tissue and can be involved in disseminated lymphoma. It highlights the importance of careful examination of WT's lymphoid stroma for the possible presence of any coexistent malignancy. PMID- 19294486 TI - Natural killer cell receptors and their ligands in liver diseases. AB - The liver is a distinctive immune organ with predominant innate immunity, being rich in innate immune cells such as natural killer (NK) cells. In humans, NK cells comprise about 30%-50% of intrahepatic lymphocytes, whereas peripheral blood lymphocytes contain about 5%-20% NK cells. Accumulating evidence suggests that NK cells play an important role not only in host defense against invading microorganisms and tumor transformation in the liver but also in liver injury and repair. In recent years, significant progress has been made in terms of understanding how NK cells recognize their target cells and carry out their effector functions. It is now clear that NK cells are strictly regulated by numerous activating and inhibitory NK cell receptors that recognize various classes of cell surface ligands, some of which are expressed by normal healthy cells. Therefore, to further elucidate the involvement of NK cells in the pathogenesis of liver diseases, an understanding of recent advances in NK cell biology is crucial. This review provides an overview of recent advances in our knowledge of human NK cell receptors and their ligands in the context of liver diseases. PMID- 19294487 TI - Interaction of a multi-domain adaptor protein, vinexin, with a Rho-effector, Rhotekin. AB - Among various effector proteins for the Rho small GTPase, the function(s) of Rhotekin is almost unknown. We have identified a multi-domain adaptor protein, vinexin, as a binding partner for Rhotekin, using yeast two-hybrid screening of a human heart library. Rhotekin was found to associate with vinexin in vitro, in COS7 cells, and in brain tissues. The C-terminal Pro-rich motif of Rhotekin exhibited binding to the third SH3 domain of vinexin. The binding was little affected by Rho but was inhibited by activated Cdc42 in COS7 cells. Immunofluorescence analyses revealed partial colocalization of vinexin-alpha with Rhotekin at focal adhesions in REF52 fibroblast cells. These results suggest that Rhotekin forms a complex with vinexin and may play a role at focal adhesions. PMID- 19294488 TI - Layer-specific sulfatide localization in rat hippocampus middle molecular layer is revealed by nanoparticle-assisted laser desorption/ionization imaging mass spectrometry. AB - Lipids are major structural component of the brain and play key roles in signaling functions in the central nervous system (CNS), such as the hippocampus. In particular, sulfatide is an abundant glycosphingolipid component of both the central and the peripheral nervous system and is an essential lipid component of myelin membranes. Lack of sulfatide is observed in myelin deformation and neurological deficits. Previous studies with antisulfatide antibody have investigated distribution of sulfatide expression in neurons; however, this method cannot distinguish the differences of sulfatide lipid species raised by difference of carbon-chain length in the ceramide portion in addition to the differences of sulfatide and seminolipid. In this study, we solved the problem by our recently developed nanoparticle-assisted laser desorption/ionization (nano PALDI)-based imaging mass spectrometry (IMS). We revealed that the level of sulfatide in the middle molecular layer was significantly higher than that in granule cell layers and the inner molecular layer in the dentate gyrus of rat hippocampus. PMID- 19294489 TI - Immunolocalization of phospho-Arg-directed protein kinase-substrate in hypoxic kidneys using in vivo cryotechnique. AB - Protein kinases (PKs) phosphorylate proteins at active regions for signal transduction. In this study, normal and hypoxic mouse kidneys were prepared using an "in vivo cryotechnique" (IVCT) and examined immunohistochemically with specific antibodies against phospho-(Ser/Thr) PKA/C substrate (P-PK-S) and phospho-(Ser/Thr) Akt substrate (P-Akt-S) to capture their time-dependent regulation in vivo. Left kidneys were cryofixed with IVCT under normal blood circulation and after varying hypoxic intervals, followed by freeze-substitution with acetone containing paraformaldehyde. Deparaffinized sections were immunostained for P-PK-S, Na(+)/HCO(3) (-) cotransporter NBC1, and a membrane skeletal protein, 4.1B. The P-PK-S was diffusely immunolocalized in the cytoplasm of the proximal tubules in normal kidneys, whereas NBC1 and 4.1B were detected at the basal striations of S1 and S2 segments of the proximal tubule. After 10 or 30 s hypoxia, P-PK-S was still immunolocalized in the cytoplasm of kidneys, but it was detected at the basal striations after 1 or 2 min hypoxia. The immunolocalization of P-Akt-S was the same as P-PK-S in the normal and hypoxic kidneys. Immunoblotting analyses of the kidney tissues under normal or hypoxic condition clearly identified the same 40-kDa bands. The IVCT is useful for time dependent analysis of the immunodistribution of P-PK-S and P-Akt-S. PMID- 19294490 TI - Apoptotic effects of Tian-Long compound on endometrial adenocarcinoma cells in vitro. AB - The Tian-Long (TL) compound is a water-soluble extract of six Chinese medicinal herbs. To explore its antitumor properties and the mechanism for activity in gynecological malignancies, the present studies were carried out using Ishikawa cells derived from uterine endometrial adenocarcinoma. Morphologically, cell death and decrease in the number of viable cells were observed in the presence of the TL compound. The proliferation of Ishikawa cells was significantly suppressed in a time- and dose-dependent manner, as indicated by both the WST-1 and the BrdU incorporation assay. Results from both the WST-1 and the BrdU incorporation assay demonstrated that the compound could inhibit the cell proliferation despite the presence of 17beta-estradiol in the medium. It is generally noted that the disturbance in mitochondrial function and DNA synthesis during cell proliferation can result in apoptosis. Being consistent with this notion, redistribution of the plasma membrane phosphatidylserine was identified with fluoromicroscopy and flow cytometry. Analysis of the fluorescent patterns of JC-1 staining revealed depolarization of the mitochondrial membrane in the exposed cells. Moreover, the amount of Bcl-2 enhanced in the presence of 17beta-estradiol was repressed by the compound. The present results indicate that the ingredients of TL compound are very promising for use in the treatment of endometrial adenocarcinoma. Further studies are necessary to elucidate the therapeutic mechanisms in its antitumor activity. PMID- 19294491 TI - Nanotransportation system for cholera toxin in Vibrio cholerae 01. AB - Vibrio cholerae (V. cholerae) cholera toxin (CT), which causes a severe watery diarrheal illness, is secreted via the type II secretion machinery; it remains unclear, however, how this toxin is transported toward the machinery. In this study, we determined that the pH-dependent intrabacterial transport system correlates with the priming of CT secretion by V. cholerae. The secretion and production of V. cholerae treated at different pHs were examined by enzyme immunoassay. The localization of the CT was analyzed by immunoelectron microscopy. The CT secretion level rapidly increases in the alkaline-pH-treated V. cholerae but does so more slowly in neutral- and acidic-pH-treated V. cholerae. The CT was found to be densely localized near the membrane in the alkaline-pH-treated bacterial cytoplasm, suggesting that the CT shifts from the center to the peripheral portion of the cytoplasm following an extracellular rise in pH. The shift was observed in V. cholerae treated at alkaline pH for more than 10 min. The pH treatment did not enhance CT production at the same stage at which secretion and intrabacterial transport of the CT were enhanced. We propose that V. cholerae possesses a pH-dependent intrabacterial nanotransportation system that probably accelerates priming for CT secretion. PMID- 19294492 TI - Ultrastructural immunolocalization of a cartilage-specific proteoglycan, aggrecan, in salivary pleomorphic adenomas. AB - A pleomorphic adenoma (PA) is the most common epithelial tumor in the salivary glands, but it frequently shows a mesenchyme-like histology, including the presence of myxoid and chondroid areas. Cartilage-specific matrix proteins are deposited in PA. Aggrecan is a major component of cartilage-specific proteoglycans. The present study examined the ultrastructure of the stromal areas in ten salivary PA specimens and investigated the distribution of aggrecan by immunoelectron microscopy. Aggrecan was deposited in the myxoid and chondroid stroma of PA. Ultrastructural observations revealed many proteoglycan cores and fibrils in the myxoid stroma and some spindle-shaped neoplastic myoepithelial cells with vacuoles and actin filaments in the myxoid areas. By immunoelectron microscopy, positivity for aggrecan was observed in the vacuoles of neoplastic myoepithelial cells, which coexisted with the viscous materials, and it was also frequently seen in electron-dense crystals in the myxoid stroma. These findings suggest that neoplastic myoepithelial cells produce aggrecan and release it from vacuoles, and aggrecan is then deposited in the myxoid stroma. Aggrecan deposition is therefore considered to play an important role in the formation of the mesenchyme-like stroma, especially the myxoid stroma. PMID- 19294493 TI - Ureteral small cell carcinoma. AB - Ureteral small cell carcinoma is very rare; only eight cases have been reported in the literature. In this article, we report the ninth case of ureteral small cell carcinoma. A 79-year-old Japanese man presented with asymptomatic macrohematuria, and left nephroureterectomy was performed. The nonpapillary and broad-based tumor, which measured 3.7 x 3.7 x 2.0 cm, was seen in the lower portion of the left ureter just above the ureteral orifice. Microscopically, the tumor was composed of urothelial carcinoma and small cell carcinoma. Immunohistochemically, neoplastic cells of small cell carcinoma were focally positive for chromogranin A. In the normal ureteral mucosa adjacent to the tumor, some endocrine cells positive for chromogranin A and synaptophysin were identified. In conclusion, we suggest that endocrine cells in the ureteral urothelial epithelium may be precursor cells of ureteral small cell carcinoma. PMID- 19294494 TI - Neuroendocrine small cell carcinoma of the breast: report of a case. AB - A rare case of neuroendocrine small cell carcinoma (SmCC) of the breast is reported. A 51-year-old postmenopausal woman noticed a nodule approximately 3 cm in diameter in her right upper breast. Histologically, the tumor consisted of small ovoid to pleomorphic cells with hyperchromatic nuclei, and a large central area was occupied by acellular amorphous tissue. Extensive lymphatic permeation was seen around the tumor. Invasive and in situ ductal carcinoma foci were not observed in and surrounding the tumor. Immunohistochemically, estrogen and progesterone receptors and HER2/neu were all negative in the tumor cells. Synaptophysin and chromogranin A were diffusely positive in the tumor cells. Cytokeratin 8 was only positive in a few tumor cells. The labeling indices of Ki 67 and p53 were high in the tumor. Postoperatively, systemic studies including positron emission tomography were performed but failed to reveal any other possible primary sites, including lung. Based on these findings, the tumor was diagnosed as neuroendocrine primary SmCC of the breast. Postoperatively, the patient received a course of weekly paclitaxel. However, pelvic bone metastasis was identified on a bone scintigram 1 year after surgery. Mammary SmCC showing high Ki-67 and p53 index should be treated carefully because of their aggressive clinical behavior. PMID- 19294495 TI - The role played by lipids unsaturation upon the membrane interaction of the Helicobacter pylori HP(2-20) antimicrobial peptide analogue HPA3. AB - The HPA3 peptide is an analogue of the linear antimicrobial peptide, HP(2-20), isolated from the N-terminal region of the Helicobacter pylori ribosomal protein, able to interact with zwitterionic lipid membranes and generate pores. Herein we focused on the importance of the degree of unsaturation of lipid acyl chains on HPA3 peptide-membrane interactions. Electrophysiology experiments carried out in reconstituted lipid membranes formed from phosphatidylcholines with one (1 palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine - POPC) and two monounsaturated acyl chains (1,2-dioleoyl-sn-glycero-3-phosphocholine - DOPC) demonstrate that the lesser degree of the packing density of membrane lipids encountered in DOPC based planar membranes greatly enhances the electric activity of pores created by the HPA3 peptide. Data derived from fluorescence spectroscopy experiments demonstrate that upon interaction with the bilayer, the HPA3 peptide translocates to the trans-side of the membrane. From the same experiments, we demonstrate that in the case of DOPC-based planar membranes, the net amount of HPA3 peptide which passes across the membrane and re-dissolves in the trans solution is almost 22% greater than POPC-based membranes. Such data further emphasize the modulatory role played by lipid acyl chain in determining antimicrobial peptides-lipids interactions, and demonstrate that small differences in unsaturation degree can impose a sizeable influence on HPA3 peptide activity. PMID- 19294496 TI - Evidence that folic acid deficiency is a major determinant of hyperhomocysteinemia in Parkinson's disease. AB - In the present work we measured blood levels of total homocysteine ((t)Hcy), vitamin B(12) and folic acid in patients with Parkinson s disease (PD) and in age matched controls and searched for possible associations between these levels with smoking, alcohol consumption, L-DOPA treatment and disease duration in PD patients. We initially observed that plasma (t)Hcy levels were increased by around 30 % in patients affected by PD compared to controls. Linear correlation, multiple regression and comparative analyses revealed that the major determinant of the increased plasma concentrations of (t)Hcy in PD patients was folic acid deficiency, whereas in controls (t)Hcy levels were mainly determined by plasma vitamin B(12) concentrations. We also observed that alcohol consumption, gender and L-DOPA treatment did not significantly alter plasma (t)Hcy, folic acid and vitamin B(12) levels in parkinsonians. Furthermore, disease duration was positively associated with (t)Hcy levels and smoking was linked with a deficit of folic acid in PD patients. Considering the potential synergistic deleterious effects of Hcy increase and folate deficiency on the central nervous system, we postulate that folic acid should be supplemented to patients affected by PD in order to normalize blood Hcy and folate levels, therefore potentially avoiding these risk factors for neurologic deterioration in this disorder. PMID- 19294497 TI - Intracerebroventricular administration of N-acetylaspartic acid impairs antioxidant defenses and promotes protein oxidation in cerebral cortex of rats. AB - N-acetylaspartic acid (NAA) is the biochemical hallmark of Canavan Disease, an inherited metabolic disease caused by deficiency of aspartoacylase activity. NAA is an immediate precursor for the enzyme-mediated biosynthesis of N acetylaspartylglutamic acid (NAAG), whose concentration is also increased in urine and cerebrospinal fluid of patients affected by CD. This neurodegenerative disorder is clinically characterized by severe mental retardation, hypotonia and macrocephaly, and generalized tonic and clonic type seizures. Considering that the mechanisms of brain damage in this disease remain not fully understood, in the present study we investigated whether intracerebroventricular administration of NAA or NAAG elicits oxidative stress in cerebral cortex of 30-day-old rats. NAA significantly reduced total radical-trapping antioxidant potential, catalase and glucose 6-phosphate dehydrogenase activities, whereas protein carbonyl content and superoxide dismutase activity were significantly enhanced. Lipid peroxidation indices and glutathione peroxidase activity were not affected by NAA. In contrast, NAAG did not alter any of the oxidative stress parameters tested. Our results indicate that intracerebroventricular administration of NAA impairs antioxidant defenses and induces oxidative damage to proteins, which could be involved in the neurotoxicity of NAA accumulation in CD patients. PMID- 19294498 TI - Trends in autism spectrum disorder diagnoses: 1994-2007. AB - We analyzed predictors of parent-reported initial diagnosis (autistic disorder [AD], pervasive developmental disorder-not otherwise specified [PDD-NOS], pervasive developmental disorder ['PDD'] and autism spectrum disorder ['ASD'], and Asperger syndrome [AS]), among 6,176 individuals with autism spectrum disorders diagnosed from 1994 through 2007. Overall, distribution of diagnoses was influenced by a secular time trend factor; other significant factors included ethnicity, white race, geographic location, urbanicity, and initial evaluator. Since 2001, most initial diagnoses of AD and AS have remained steady while 'PDD' and PDD-NOS have decreased. 'ASD' diagnoses have increased, especially among school-based teams; AS diagnoses also increased uniquely among these evaluators. Findings from this study suggest that current diagnostic guidelines may not be meeting all community evaluator needs. PMID- 19294499 TI - Non-structural proteins of Periplaneta fuliginosa densovirus inhibit cellular gene expression and induce necrosis in Sf9 cell cultures. AB - The non-structural protein NS1 of Periplaneta fuliginosa densovirus (PfDNV) is a multifunctional protein that has previously been shown to possess ATP-binding, ATPase, site-specific DNA-binding, helicase, and transcription activation activities. We report here an investigation of the cytopathogenicity of this viral non-structural (NS) protein, as well as other two NSs, NS2, and NS3, in cultured insect cells. The expression of NS1 alone potently inhibited cellular gene expression, whereas NS2 and NS3 did not produce a similar effect. The inhibition of gene expression by NS1 was confirmed to be specific and not a simple manifestation of toxicity. For example, NS1 inhibited expression of several reporter genes under the control of different RNA polymerase II promoters, whereas it did not inhibit expression from a T7 RNA polymerase promoter construct. Mapping analysis identified the carboxy-terminal peptide of this protein as the region important for the inhibition of cellular gene expression, suggesting that this inhibition is independent of its DNA-binding activity. Next, the mutagenesis assay showed that ATP-binding was essential for the unique function of this protein. Furthermore, we found that NS2 and NS3 cooperatively enhanced the NS1-induced transcription inhibition. Co-expression of all the three NS proteins in Sf9 cells also led to necrotic cell death by ATP depletion. PMID- 19294500 TI - Microfinance and HIV/AIDS prevention: assessing its promise and limitations. AB - Researchers increasingly argue that poverty and gender inequality exacerbate the spread of HIV/AIDS and that economic empowerment can therefore assist in the prevention and mitigation of the disease, particularly for women. This paper critically evaluates such claims. First, we examine the promises and limits of integrated HIV/AIDS prevention and microfinance programs by examining the available evidence base. We then propose future research agendas and next steps that may help to clear current ambiguities about the potential for economic programs to contribute to HIV/AIDS risk reduction efforts. PMID- 19294501 TI - Tolerability of and adherence to combination oral therapy with gefitinib and capecitabine in metastatic breast cancer. AB - PURPOSE: This phase I study explored gefitinib (G) and capecitabine (C) in metastatic breast cancer (MBC). METHODS: Sequential cohorts (n = 3) received G and escalating C on a 14 day on/7 day off schedule, with a validation cohort (n = 10) at the maximum tolerated dose (MTD). Dose limiting toxicity (DLT) was defined in cycle 1. The primary endpoint was safety; secondary endpoints included response and adherence. RESULTS: About 19 patients were treated for a median of 5 cycles. No patients in sequential cohorts experienced DLT; C MTD was 2,000 mg/m(2)/day when paired with daily G 250 mg. In the validation cohort, four experienced serious toxicities, including diarrhea, mucositis, and palmarplantar dysesthesia. At the MTD, 6 (46%) required a C dose reduction, and 3 (23%) came off study for toxicity. One partial response was observed (8%, 95% CI 0.2-38.5%); five had stable disease >24 weeks (26, 95% CI 9-51%). Patients missed few drug doses, with the suggestion of overadherence to therapy. CONCLUSIONS: In this phase I study of G and C in MBC, a C MTD was identified, and significant toxicity was observed. About 8% demonstrated a response, with 26% maintaining stable disease. The possibility of overadherence, as suggested in this study, may have implications for other trials of oral antineoplastic therapy. PMID- 19294502 TI - The distribution over time of costs and social net benefits for pertussis immunization programs. AB - The cost of a six-dose pertussis immunization programs for children and adolescents is investigated in relation to estimators of the price of acellular vaccine, the value of a child's life, levels of vaccination rate and discount rates. We compare the cost of the program maintained over time at 90% with three alternative strategies, each involving a decrease in vaccination coverage. Data from England and Wales, 1966-2005, is used to formalize a delay in occurrence of pertussis cases as a result of a fall in coverage. We first apply the criterion of minimization of the total social cost of pertussis to identify the best cost saving immunization strategy. The results are also discussed in form of the discounted present value of the total social net benefits. We find that the discounted present value of the total social net benefit is maximized when a stable vaccination program at 90% is compared to a gradual decrease in vaccination coverage leading to the lowest vaccination rate. The benefits to society of providing sustained immunization strategy, vaccinating the highest proportion of children and adolescents, are systematically proved on the basis of the second optimisation criterion, independently of the level of estimators applied during economic evaluation for the cost variables. PMID- 19294503 TI - Neuroprotective effect of baicalin in a rat model of permanent focal cerebral ischemia. AB - This investigation was performed to determine the neuroprotective effect of baicalin on permanent cerebral ischemia injury in rats and the potential mechanisms in this process. Adult male Sprague-Dawley rats underwent permanent middle cerebral artery occlusion (pMCAO). The rats were then received intraperitoneal injection with baicalin (10, 30 and 100 mg/kg) or vehicle. Morphological characteristic, neurological deficit scores, cerebral infarct volume and the enzymatic activity of myeloperoxidase (MPO) were measured 24 h after pMCAO. The mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were determined by RT-PCR. Neuronal apoptosis was determined by TUNEL staining and Western blot. Baicalin (30 and 100 mg/kg) reduced neurological deficit scores and cerebral infarct volume 24 h after pMCAO. Baicalin significantly decreased the enzymatic activity of MPO and the expression of iNOS mRNA and COX-2 mRNA in rat brain, it also significantly inhibited neuronal apoptosis and the expression of cleaved caspase-3 protein after pMCAO. Our results suggested that baicalin possesses potent anti-inflammatory and anti apoptotic properties and attenuates cerebral ischemia injury. This protection might be associated with the downregulated expression of iNOS mRNA, COX-2 mRNA, and cleaved caspase-3 protein. PMID- 19294504 TI - Impact of a living donor liver transplant program for hepatocellular carcinoma to the running local deceased donor liver transplant program. PMID- 19294505 TI - Genetic association of nonsynonymous variants of the IL23R with familial and sporadic inflammatory bowel disease in women. AB - PURPOSE: To replicate the association of IL23R R381Q (rs11209026) with inflammatory bowel disease (IBD), examine the effect of the two nonsynonymous variations, Q3H and L310P, on IBD, and to study gender distribution of these variants in IBD patients. RESULTS: IL23R R381Q was associated with Crohn's disease (CD) (P = 0.010), but not with ulcerative colitis (UC); L310P was associated with UC (P = 0.004), but not with CD; no association was observed for Q3H with CD or UC. A female-specific association of R381Q with CD (P = 0.041), and of L310P with UC (P = 0.008) was observed. CONCLUSION: We replicated the association of IL23R R381Q with CD but not UC, and we observed an association of L310P with UC, but not CD, in a central Pennsylvania population. Further analysis of the distribution of IL23R variants revealed that these effects were largely female-specific. The results suggest that IL23R R381Q confers protection against CD and that L310P confers protection against UC in females. PMID- 19294506 TI - The clinical value of serum connective tissue growth factor in the assessment of liver fibrosis. AB - To explore the relation between connective tissue growth factor (CTGF) in serum and the severity of liver fibrosis, and to determine the clinical value of CTGF in the assessment of liver fibrosis, serum CTGF was tested utilizing enzyme linked immunosorbent assay (ELISA). The correlation between serum CTGF concentration and fibrosis stage was assessed. The diagnostic performance of CTGF was assessed by comparing the area under the receiver operating characteristic (ROC) curves (AUC) with a panel of fibrosis markers. The correlation coefficient was 0.689 (P < 0.001) between the levels of serum CTGF and fibrosis stages and the AUC of CTGF was 0.841 (95% confidence interval [CI] 0.762-0.920) in distinguishing mild fibrosis from significant fibrosis. The present data revealed that serum CTGF was significantly correlated with the stage of liver fibrosis, suggested that serum CTGF was an indicator for the stage of liver fibrosis, and shown evidence that serum CTGF could be used as a valuable marker for assessing liver fibrosis. PMID- 19294507 TI - Preventive effect of pentoxifylline on acute radiation damage via antioxidant and anti-inflammatory pathways. AB - PURPOSE: The aim of the present study was to investigate whether pentoxifylline (PTX) treatment could protect against induced acute radiation enteritis. METHOD: Rats received 100 mg/kg/day PTX for 7 days before irradiation and continued on treatment for 3 days after irradiation. The intestinal myeloperoxidase (MPO) activities and malondialdehyde (MDA), glutathione (GSH), prostaglandin E2, and thromboxane B2 levels were determined. Terminal ileum tissue was evaluated for morphological changes. Also, nuclear factor kappa (NF-kappa), tumor necrosis factor-alpha (TNF-alpha), and intercellular adhesion molecule 1 (ICAM-1) expressions were analyzed with immunohistochemisty methods. RESULTS: PTX treatment was associated with increased GSH levels and decreased MPO activity and MDA, prostaglandin E2, and thromboxane B2 levels. Histopathologic examination showed that intestinal mucosal structure was preserved in the PTX-treated group while having significant decreases in NF-kappaB, TNF-a, and ICAM-1 expression. CONCLUSIONS: PTX appears to have a protective effect against radiation damage. This protective effect is mediated in part by decreasing both inflammatory reactions and oxidative stress. PMID- 19294508 TI - Wireless video capsule enteroscopy in preclinical studies: methodical design of its applicability in experimental pigs. AB - The aim of this project was to develop a methodology to introduce wireless video capsule endoscopy in preclinical research. Five mature female pigs (Sus scrofa domestica) were selected for the study. Capsule endoscopes (the EndoCapsule system; Olympus) were introduced into the duodenum endoscopically in each of the animals. The life span of batteries (i.e., total time of endoscopy recording) was 487-540 min (median 492 min). The capsule endoscope reached the cecum during enteroscopy once (after 7 h 57 min), in the remaining cases, endoscopy recordings terminated in the distal or terminal ileum. All capsule enteroscopies found a normal pattern of the small intestine. The intestinal lumen is narrower, transverse folds are sparse or even absent, villi are wider but less prominent in pigs compared to humans. Capsule endoscopy in experimental pigs will be helpful for future trials on injury of different drugs and xenobiotics to the small bowel. PMID- 19294509 TI - Evaluating breath methane as a diagnostic test for constipation-predominant IBS. AB - Studies suggest that subjects with IBS have altered gut flora. Among these findings, methane production is more commonly associated with constipation predominant symptoms. In this study, we prospectively evaluated the role of methane as a diagnostic test. Consecutive Rome I positive IBS patients referred for a lactulose breath test were eligible to participate. After exclusion criteria, subjects completed a symptom questionnaire grading bloating, diarrhea, and constipation on a VAS scale (0-100 mm). Once completed, a physician interviewed the subjects and rated the subject accordingly, and also determined whether the patient had C-IBS, D-IBS, or neither. Subjects and physicians were blinded to the results of the breath test. The presence of methane in the breath test was compared to the results of the scoring by subjects and physicians. A total of 56 Rome I positive IBS subjects were enrolled. During breath testing, 28 subjects produced methane. Good agreement between physician's evaluation and the patient's was seen (diarrhea = 0.69; constipation = 0.69; bloating = 0.62). The severity of constipation was noted to be greater in the methane group (49.3 +/- 28.7) than in the non-methane group (25.3 +/- 31.47) (P < 0.01). In contrast, diarrhea was less severe in the methane group (12.3 +/- 21.0) than the non methane group (36.7 +/- 32.4) (P < 0.01). Out of the 56 patients, 23 C-IBS subjects were identified by the physician. When methane was used to predict the assignment of C-IBS compared to non-C-IBS, it had a sensitivity of 91.7% and a specificity of 81.3% (OR = 47.7, CI = 9.4-232, P < 0.00001). In conclusion, methane is a potential diagnostic test for the identification of C-IBS and may guide treatment. PMID- 19294510 TI - Are the echogenicities on intraductal ultrasonography really biliary microlithiasis? AB - BACKGROUND: Intraductal ultrasonography (IDUS) is a useful procedure for diagnosing microlithiasis in the bile duct but it is not easy to differentiate between tiny echogenicity and real microlithiasis. We compared the echogenicity seen on IDUS and the findings of bile microscopy (BM) of bile that was collected in the common bile duct (CBD) to determine whether the echogenicity seen on IDUS is real microlithiasis. METHODS: This prospective study involved a total of 30 patients who experienced biliary pain (n = 11), acute cholecystitis (n = 11) or indeterminate pancreatitis (n = 8) without a filling defect or obstruction in the bile duct. IDUS was performed during endoscopic retrograde cholangiopancreatography (ERCP), followed by bile aspiration for BM. Endoscopic sphincterotomy (EST) was performed if definite echogenic materials were observed on IDUS. RESULTS: Of the 30 patients, 23 (77%) had echogenic materials visible in the CBD on IDUS. Of these 23 patients, 13 (57%) were found to have biliary crystals by BM. The size of the echogenic materials was the only significant factor associated with BM positivity. Using the receiver operating curve, the optimal size of the echogenicity to differentiate real microlithiasis was 1.4 mm. CONCLUSIONS: Optimal concordance between IDUS and BM was observed when the size of the microlithiasis was greater than 1.4 mm; under these conditions the sensitivity and specificity were 71% and 75%, respectively. This information may be useful when deciding whether to perform endoscopic sphincterotomy. PMID- 19294511 TI - G-CSF enhanced SDF-1 gradient between bone marrow and liver associated with mobilization of peripheral blood CD34+ cells in rats with acute liver failure. AB - The role of stromal cell-derived factor-1 (SDF-1) in modulating massive liver damage is not well known. In this study, expression of SDF-1 in bone marrow and liver was investigated in rats with acute liver failure (ALF) when mobilized using granulocyte colony-stimulating factor (G-CSF). ALF was induced in rats by D galactosamine (D-GalN). Starting after 2 hours following D-GalN induction, the animals were injected with G-CSF 50 microg/kg daily or saline as placebo for 5 days. The percentages of CD34+ cells in peripheral blood and the expression of SDF-1 in bone marrow and liver were then determined. The percentages of peripheral CD34+ cells demonstrated a transient increase in placebo rats following D-GalN induction and a significant increase in rats after G-CSF administration. SDF-1 expression showed a transient decrease in bone marrow and a transient increase in liver tissue from placebo rats. However, a significant decrease of SDF-1 expression in bone marrow and a remarkable increase in liver tissue were observed in animals from the G-CSF group. It was concluded that G-CSF can enhance the reduced expression of SDF-1 in bone marrow and increased expression in liver in ALF rats, forming a greater SDF-1 gradient, and chemoattracting CD34+ cells' migration from bone marrow to an injured liver. PMID- 19294512 TI - Correlates of bacterial pneumonia hospitalizations in elders, Texas border. AB - BACKGROUND: Immunization preventable bacterial pneumonia is an Agency for Healthcare Research and Quality (AHRQ) prevention quality indicator of health care. This study explored associations of individual and county correlates with bacterial pneumonia hospitalization rates for elders residing in 32 Texas counties bordering Mexico. METHODS: We estimated baseline rates from Texas Health Care Information Collection's hospital discharge data for 1999-2001, and population counts from the 2000 U.S. Census. RESULTS: The rate among the total Texas border population was 500/10,000, three times the national rate. Elders 75+, males, and Latinos had the highest rates. An increase of 1 primary care physician per 1000 population is associated with a decrease in pneumonia-related hospitalization rates by 33%, while each 10% increase in Latinos is associated with a 0.1% rate increase. DISCUSSION: This baseline bacterial pneumonia hospitalization study demonstrates a systematic approach to estimate county rates, a process that could lead to improved outcomes through effective community interventions. Methodology demonstrates how publicly available hospital discharge data can be used by communities to better measure and improve quality of health care. PMID- 19294513 TI - Optimal control of HIV-virus dynamics. AB - In this paper we consider a mathematical model of HIV-virus dynamics and propose an efficient control strategy to keep the number of HIV virons under a pre specified level and to reduce the total amount of medications that patients receive. The model considered is a nonlinear third-order model. The third-order model describes dynamics of three most dominant variables: number of healthy white blood cells (T-cells), number of infected T-cells, and number of virus particles. There are two control variables in this model corresponding to two categories of antiviral drugs: reverse transcriptase inhibitors (RTI) and protease inhibitors (PI). The proposed strategy is based on linearization of the nonlinear model at the equilibrium point (steady state). The corresponding controller has two components: the first one that keeps the system state variables at the desired equilibrium (set-point controller) and the second-one that reduces in an optimal way deviations of the system state variables from their desired equilibrium values. The second controller is based on minimization of the square of the error between the actual and desired (equilibrium) values for the linearized system (linear-quadratic optimal controller). The obtained control strategy recommends to HIV researchers and experimentalists that the constant dosages of drugs have to be administrated at all times (set point controller, open-loop controller) and that the variable dosages of drugs have to be administrated on a daily basis (closed-loop controller, feedback controller). PMID- 19294514 TI - Novel biological/biohybrid prostheses for the ossicular chain: fabrication feasibility and preliminary functional characterization. AB - Alternatives for ossicular replacements were fabricated in order to overcome persisting rejections in middle ear prosthetization. Unlike the synthetic prostheses in fashion, we propose biological and biohybrid replacements containing extra cellular matrix (ECM) molecules to improve biointegration. In this study, ECM-containing devices shaped as Partial Ossicular Replacement Prostheses (PORPs) were fabricated reproducing the current synthetic models. Biological PORPs were obtained from human decellularized cortical bone allografts by computer numerically controlled ultraprecision micromilling. Moreover, porous PORP-like scaffolds were produced and cultured with osteoinduced human mesenchymal stromal cells to generate in vitro bone ECM within the scaffold porosity (biohybrid PORPs). The acoustic responses of such devices were investigated and compared to those of commercial prostheses. Results showed that biological PORPs transmit mechanical signals with appropriate frequencies, amplitudes, and with early extinction time. Although signal transmission in biohybrid PORPs showed insufficient amplitude, we believe that tissue engineered constructs represent the new challenge in ossiculoplasty. PMID- 19294515 TI - Race-norming of neuropsychological tests. AB - Recent studies in the United States indicate that some neurologically intact minority groupings perform well below White Americans on neuropsychological tests. This has sparked the production of race-norms, especially for African Americans, that seek to reduce false positive rates (i.e., neurologically intact individuals misdiagnosed with cognitive impairment) in neuropsychological assessments. There are problems with this enterprise including: possible justification for inferior/superior treatment of different racial groupings; unknown effects on false negative rates (i.e., cognitive deficit misdiagnosed as normal); the overlooking of factors possibly responsible for group racial differences (e.g., acculturation); non-scientific and non-operational definitions of race/ethnic groupings; and an impossibly large number of potential race/ethnic groupings for which to generate race-norms. An alternative approach is to use a single set of combined race/ethnic norms and estimate preexisting neuropsychological skill levels by using individual comparison standards. This alternative has been poorly researched, a situation that needs correcting. PMID- 19294516 TI - Differential diagnosis of ACTH-dependent hypercortisolism: imaging versus laboratory. AB - Differential diagnosis of ACTH-dependent Cushing's syndrome often presents major difficulties. Diagnostic troubles are increased by suboptimal specificity of endocrine tests, the rarity of ectopic ACTH secretion and the frequent incidental discovery of pituitary adenomas. A 43-year-old female reported with mild signs and symptoms of hypercortisolism, and initial hormonal tests and results of pituitary imaging (7-mm adenoma) were suggestive for Cushing's disease. However, inadequate response to corticotrophin-releasing hormone and failure to suppress after 8 mg dexamethasone pointed towards an ectopic source. Total body CT scan visualized only a small, non-specific nodule in the right posterior costophrenic excavation. Inferior petrosal sinus sampling revealed an absent center:periphery ACTH gradient but octreoscan and (18)F-FDG-PET-CT failed to detect abnormal tracer accumulation. We weighed results of the laboratory with those of imaging and decided to remove the lung nodule. Pathology identified a typical, ACTH staining carcinoid and the diagnosis was confirmed by postsurgical hypoadrenalism. In conclusion, imaging may prove unsatisfactory or even misleading for the etiologial diagnosis of ACTH-dependent Cushing's syndrome and should therefore be interpreted only in context with results of hormonal dynamic testing. PMID- 19294517 TI - Preparation and evaluation of miconazole nitrate-loaded solid lipid nanoparticles for topical delivery. AB - The purpose of this study was to prepare miconazole nitrate (MN) loaded solid lipid nanoparticles (MN-SLN) effective for topical delivery of miconazole nitrate. Compritol 888 ATO as lipid, propylene glycol (PG) to increase drug solubility in lipid, tween 80, and glyceryl monostearate were used as the surfactants to stabilize SLN dispersion in the SLN preparation using hot homogenization method. SLN dispersions exhibited average size between 244 and 766 nm. All the dispersions had high entrapment efficiency ranging from 80% to 100%. The MN-SLN dispersion which showed good stability for a period of 1 month was selected. This MN-SLN was characterized for particle size, entrapment efficiency, and X-ray diffraction. The penetration of miconazole nitrate from the gel formulated using selected MN-SLN dispersion as into cadaver skins was evaluated ex-vivo using franz diffusion cell. The results of differential scanning calorimetry (DSC) showed that MN was dispersed in SLN in an amorphous state. The MN-SLN formulations could significantly increase the accumulative uptake of MN in skin over the marketed gel and showed a significantly enhanced skin targeting effect. These results indicate that the studied MN-SLN formulation with skin targeting may be a promising carrier for topical delivery of miconazole nitrate. PMID- 19294518 TI - Integrated primary care: an inclusive three-world view through process metrics and empirical discrimination. AB - Integrating behavioral health services within the primary care setting drives higher levels of collaborative care, and is proving to be an essential part of the solution for our struggling American healthcare system. However, justification for implementing and sustaining integrated and collaborative care has shown to be a formidable task. In an attempt to move beyond conflicting terminology found in the literature, we delineate terms and suggest a standardized nomenclature. Further, we maintain that addressing the three principal worlds of healthcare (clinical, operational, financial) is requisite in making sense of the spectrum of available implementations and ultimately transitioning collaborative care into the mainstream. Using a model that deconstructs process metrics into factors/barriers and generalizes behavioral health provider roles into major categories provides a framework to empirically discriminate between implementations across specific settings. This approach offers practical guidelines for care sites implementing integrated and collaborative care and defines a research framework to produce the evidence required for the aforementioned clinical, operational and financial worlds of this important movement. PMID- 19294519 TI - Measures of physical activity and their correlates: the Swedish National March Cohort. AB - We compared the results of self-estimates of physical activity obtained with a novel instrument (the Energy Expenditure Questionnaire, EEQ) to those obtained from questions typically asked in epidemiological investigations (reference method) in a cohort of 42,150 Swedish men and women, aged 18-94. In the EEQ, participants were asked to report total physical activity by estimating the total time during a typical day and night spent on different physical activity intensity levels from the lowest (corresponding to lying in bed, 0.9 Metabolic Energy Turnover; MET) to the highest, (exceeding the intensity of to shovelling snow by hand, i.e., >6 MET). As a comparison, they also estimated hours per week devoted to household chores, commuting and leisure time physical activities classified as; light, moderate and heavy. The average physical activity estimated with the EEQ was 1.36 MET or 32.6 METh/day or 2,341 kcal/day. In comparison, physical activity estimated with the reference method represented no more than 17% of this amount. The Spearman's correlation coefficient between the two measures was 0.26. Using EEQ, men reported significantly more physical activity than women (mean = 36.3 vs. 30.6 METh/day). Body mass index (BMI) > or = 25, education > or = 12 years, and age > or = 60 years were significantly associated with lower physical activity. Questions focusing on leisure time exercise and a few other selected activities provide estimates of activity that correlate poorly with self-reported total energy output from all physical activity and inactivity. Investigators need to be more explicit about which component of activity they investigate. PMID- 19294520 TI - Inhibition of mammary tumor growth and metastases to bone and liver by dietary grape polyphenols. AB - The cancer preventive properties of grape products such as red wine have been attributed to polyphenols enriched in red wine. However, much of the studies on cancer preventive mechanisms of grape polyphenols have been conducted with individual compounds at concentrations too high to be achieved via dietary consumption. We recently reported that combined grape polyphenols at physiologically relevant concentrations are more effective than individual compounds at inhibition of ERalpha(-), ERbeta(+) MDA-MB-231 breast cancer cell proliferation, cell cycle progression, and primary mammary tumor growth (Schlachterman et al., Transl Oncol 1:19-27, 2008). Herein, we show that combined grape polyphenols induce apoptosis and are more effective than individual resveratrol, quercetin, or catechin at inhibition of cell proliferation, cell cycle progression, and cell migration in the highly metastatic ER (-) MDA-MB-435 cell line. The combined effect of dietary grape polyphenols (5 mg/kg each resveratrol, quercetin, and catechin) was tested on progression of mammary tumors in nude mice created from green fluorescent protein-tagged MDA-MB-435 bone metastatic variant. Fluorescence image analysis of primary tumor growth demonstrated a statistically significant decrease in tumor area by dietary grape polyphenols. Molecular analysis of excised tumors demonstrated that reduced mammary tumor growth may be due to upregulation of FOXO1 (forkhead box O1) and NFKBIA (IkappaBalpha), thus activating apoptosis and potentially inhibiting NfkappaB (nuclear factor kappaB) activity. Image analysis of distant organs for metastases demonstrated that grape polyphenols reduced metastasis especially to liver and bone. Overall, these results indicate that combined dietary grape polyphenols are effective at inhibition of mammary tumor growth and site-specific metastasis. PMID- 19294521 TI - Osteoblast-induced EGFR/ERBB2 signaling in androgen-sensitive prostate carcinoma cells characterized by multiplex kinase activity profiling. AB - Bone metastases in prostate cancer are predominantly osteoblastic. To study regulatory mechanisms underlying the establishment of prostate cancer within an osteoblastic microenvironment, human androgen-sensitive prostate carcinoma cells (LNCaP) were treated with culture medium conditioned by human osteoblast-derived sarcoma cells (OHS), and activated signalling pathways in the carcinoma cells were analyzed using microarrays with tyrosine kinase substrates. Network interaction analysis of substrates with significantly increased phosphorylation levels revealed that signalling pathways mediated by EGFR and ERBB2 were activated in LNCaP cells under OHS influence but also by androgen treatment. Activation of EGFR/ERBB2 signalling was also found in LNCaP cells in cocultures with OHS cells or osteoblastic cells that had been differentiated from human mesenchymal stem cells. Our experimental data suggests osteoblast-directed induction of signalling activity via EGFR and ERBB2 in prostate carcinoma cells and may provide a rationale for the use of EGFR or ERBB2 inhibition in systemic prevention or treatment of metastatic prostate cancer in the androgen-sensitive stage of the disease. PMID- 19294522 TI - Relationship between cardiopulmonary mortality and cancer risk and quantitative exposure to polycyclic aromatic hydrocarbons, fluorides, and dust in two prebake aluminum smelters. AB - OBJECTIVES: We examined the risk of mortality and cancer incidence with quantitative exposure to benzene-soluble fraction (BSF), benzo(a)pyrene (BaP), fluoride, and inhalable dust in two Australian prebake smelters. METHODS: A total of 4,316 male smelter workers were linked to mortality and cancer incidence registries and followed from 1983 through 2002 (mean follow-up: 15.9 years, maximum: 20 years). Internal comparisons using Poisson regression were undertaken based on quantitative exposure levels. RESULTS: Smoking-adjusted, monotonic relationships were observed between respiratory cancer and cumulative inhalable dust exposure (trend p = 0.1), cumulative fluoride exposure (p = 0.1), and cumulative BaP exposure (p = 0.2). The exposure-response trends were stronger when examined across the exposed categories (BaP p = 0.1; inhalable dust p = 0.04). A monotonic, but not statistically significant trend was observed between cumulative BaP exposure and stomach cancer (n = 14). Bladder cancer was not associated with BaP or BSF exposure. No other cancer and no mortality outcomes were associated with these smelter exposures. CONCLUSIONS: The carcinogenicity of Soderberg smelter exposures is well established; in these prebake smelters we observed an association between smelter exposures and respiratory cancer, but not bladder cancer. The exploratory finding for stomach cancer needs confirmation. These results are preliminary due to the young cohort and short follow-up time. PMID- 19294523 TI - Pleural and peritoneal mesotheliomas in SEER: age effects and temporal trends, 1973-2005. AB - We analyzed mesothelioma incidence in the Surveillance, Epidemiology, and End Results (SEER) database over the period 1973-2005 using extensions of the age period-cohort (APC) models. In these analyses, the usual non-specific age effects of the conventional APC models were replaced by hazard functions derived from two multistage models of carcinogenesis, the Armitage-Doll model and the two-stage clonal expansion (TSCE) model. The extended APC models described the incidence data on pleural and peritoneal mesotheliomas well. After adjustment for temporal trends, the data suggest that the age-specific incidence rates of both pleural and peritoneal mesotheliomas are identical in men and women. Driven largely by birth cohort effects, age-adjusted rates of pleural mesothelioma among men rose from about 7.5 per million person-years in 1973 to about 20 per million person years in the early 1990s and appear to be stable or declining thereafter. Age adjusted rates of pleural mesothelioma among women have remained more or less constant at about 2.5 per million person-years over the period 1973-2005. Age adjusted rates for peritoneal mesothelioma in both men (1.2 per million person years) and women (0.8 per million person-years) exhibit no temporal trends over the period of the study. We estimate that approximately 94,000 cases of pleural and 15,000 cases of peritoneal mesothelioma will occur in the US over the period 2005-2050. PMID- 19294524 TI - Development of groundwater quality index. AB - Assessing the water quality status for special use is the main objective of any water quality monitoring studies. The water quality index (WQI) is a mathematical instrument used to transform large quantities of water quality data into a single number which represents the water quality level. In fact, developing WQI in an area is a fundamental process in the planning of land use and water resources management. In this study, a simple methodology based on multivariate analysis is developed to create a groundwater quality index (GWQI), with the aim of identifying places with best quality for drinking within the Qazvin province, west central of Iran. The methodology is based on the definition of GWQI using average value of eight cation and anion parameters for 163 wells during a 3-year period. The proportion of observed concentrations to the maximum allowable concentration is calculated as normalized value of each parameter in observing wells. Final indices for each well are calculated considering weight of each parameter. In order to assess the groundwater quality of study area, the derived indices are compared with those of well-known mineral waters. Using developed indices, groundwater iso-index map for study area and the map of areas of which the indices are near to mineral waters was drawn. In the case study, the GWQI map reveals that groundwater quality in two areas is extremely near to mineral water quality. Created index map provides a comprehensive picture of easily interpretable for regional decision makers for better planning and management. PMID- 19294525 TI - Geochemical characterisation of major and trace elements in the coastal sediments of India. AB - Thirty-five surface sediment samples from the Indian continental shelf were recovered offshore from the mouths of the major rivers (Brahmaputra, Ganges, Narmada, Tapti, Godavari, Krishna and Cauvery) discharging into the coastal region of both east and west coasts were analysed using inductively coupled plasma atomic emission spectroscopy for selected major (i.e. Al, Ca, Fe, K, Ti, Mg and Na) and trace elements (e.g. Ba, Co, Cr, Cu, Ga, Ni, P and V), after total dissolution. The main objectives are to understand the processes controlling major and trace elements in the surface sediments and to identify natural and anthropogenic sources in the coastal environment using statistically regressed elemental concentrations to establish regional baseline levels. Metal enrichments observed close to the major urban areas in the east and west coasts are associated with the industrialised activities areas rich in Cu and Co in both the east and west coast sediments. Normalisation of metals to Al indicated that high enrichment factors are in the order of Ca > Ti > or = Fe > Na > Mg > Co > Cu > Ga > V > Ba except K and P depletion. This indicated that the characteristic of estuarine sediment showed higher level along the west coast of India, which was reflected in the coastal sediments as similar to the source of its origin from the riverine composition and its abundances. PMID- 19294526 TI - Spectrofluorimetric study of the interaction of methyl-parathion with fish serum albumin. AB - The interaction of methyl-parathion with the albumin of Piaractus mesopotamicus (Holmberg 1887) (= pacu), a fish species typical of Brazilian rivers, was studied and the results compared with known values for human and bovine albumin obtained in an earlier investigation. Methyl-parathion (O,O-dimethyl O-p-nitrophenyl phosphorothioate) is an organophosphorous pesticide still used in agriculture and fish farming in many countries. The fluorescence quenching technique with tryptophan as a natural probe was used to detect for the presence of methyl parathion. Fluorescence can be mathematically expressed by the Stern-Volmer equation to calculate quenching constants, and changes in the behavior of Stern Volmer curves at different temperatures indicate the nature of the mechanism causing the quenching. Our results indicate that methyl-parathion forms a complex with fish albumin. The estimated association constant is 9.73 x 103 (+/- 4.9 x 102) M(-1) at 25 degrees C. PMID- 19294527 TI - Formation of embryoid bodies from mouse embryonic stem cells cultured on silicon coated surfaces. AB - Embryoid bodies (EBs) are primitive embryonic structures derived from differentiating embryonic stem cells (ESCs). Many techniques have been used to obtain EBs. Improving the technique of EB formation can help in achieving better results in ESCs differentiation into neurons, myocardiocytes, haemopoeitic cells, and others. We evaluated the use of Sigmacote as a hydrophobic substrate to improve EB formation. CCE and P19 cell lines were used to obtain EBs and retinoic acid was used to induce neural differentiation. The results revealed that Sigmacote, as a hydrophobic substrate, can improve EB formation from ESCs. Our results demonstrate that the silicon-coating of glass petri dishes by Sigmacote is an easy and reproducible technique to enhance EB formation from murine ESCs and EC cells. PMID- 19294528 TI - Phenotypic and molecular characterization of two novel CTX-M enzymes carried by Klebsiella pneumoniae. AB - Two clinical strains (Klebsiella pneumoniae 516 and K. pneumoniae 1335) collected in September 2006 from different hospitals in Anhui Province (China) harboured two novel plasmid-mediated bla(CTX-M) genes, designated bla(CTX-M-80) and bla(CTX M-81), respectively. Both CTX-M-80 with pI of 9.0 and CTX-M-81 with pI of 8.4 were extended-spectrum beta-lactamases (ESBLs). The results of susceptibility testing demonstrated two enzymes were highly activity against broad spectrum beta lactams, but the level of resistance was reduced with the addition of beta lactamase inhibitors. The bla(CTX-M-80) gene was detected on a 110-kb plasmid and the bla(CTX-M-81) gene existed on a 120-kb plasmid. The deduced amino acid sequence of CTX-M-80 differed from that of CTX-M-3 by the substitution Ala-27- >Val, and CTX-M-81 possessed the Lys-->Glu, Lys-->Gln, and Asn-->His changes at respective position 82, 98, and 132 in compassion with CTX-M-14. The enzymatic properties showed CTX-M-80 and CTX-M-81 had higher affinities for penicillin G (lower Km values) than for cephalosporins. The activities of novel enzymes against ceftazidime were undetectable or limited, as indicated by MICs data, the same response being observed for many other CTX-M enzymes. This report was evidence of the diversity of CTX-M-type ESBLs in China. PMID- 19294529 TI - Having a fit: impact of number of items and distribution of data on traditional criteria for assessing IRT's unidimensionality assumption. AB - PURPOSE: Confirmatory factor analysis fit criteria typically are used to evaluate the unidimensionality of item banks. This study explored the degree to which the values of these statistics are affected by two characteristics of item banks developed to measure health outcomes: large numbers of items and nonnormal data. METHODS: Analyses were conducted on simulated and observed data. Observed data were responses to the Patient-Reported Outcome Measurement Information System (PROMIS) Pain Impact Item Bank. Simulated data fit the graded response model and conformed to a normal distribution or mirrored the distribution of the observed data. Confirmatory factor analyses (CFA), parallel analysis, and bifactor analysis were conducted. RESULTS: CFA fit values were found to be sensitive to data distribution and number of items. In some instances impact of distribution and item number was quite large. CONCLUSIONS: We concluded that using traditional cutoffs and standards for CFA fit statistics is not recommended for establishing unidimensionality of item banks. An investigative approach is favored over reliance on published criteria. We found bifactor analysis to be appealing in this regard because it allows evaluation of the relative impact of secondary dimensions. In addition to these methodological conclusions, we judged the items of the PROMIS Pain Impact bank to be sufficiently unidimensional for item response theory (IRT) modeling. PMID- 19294530 TI - ATP, P2 receptors and the renal microcirculation. AB - Purinoceptors are rapidly becoming recognised as important regulators of tissue and organ function. Renal expression of P2 receptors is broad and diverse, as reflected by the fact that P2 receptors have been identified in virtually every major tubular/vascular element. While P2 receptor expression by these renal structures is recognised, the physiological functions that they serve remains to be clarified. Renal vascular P2 receptor expression is complex and poorly understood. Evidence suggests that different complements of P2 receptors are expressed by individual renal vascular segments. This unique distribution has given rise to the postulate that P2 receptors are important for renal vascular function, including regulation of preglomerular resistance and autoregulatory behaviour. More recent studies have also uncovered evidence that hypertension reduces renal vascular reactivity to P2 receptor stimulation in concert with compromised autoregulatory capability. This review will consolidate findings related to the role of P2 receptors in regulating renal microvascular function and will present areas of controversy related to the respective roles of ATP and adenosine in autoregulatory resistance adjustments. PMID- 19294531 TI - Strategies for blocking the fibrogenic actions of connective tissue growth factor (CCN2): From pharmacological inhibition in vitro to targeted siRNA therapy in vivo. AB - Connective tissue growth factor (CCN2) is a major pro-fibrotic factor that frequently acts downstream of transforming growth factor beta (TGF-beta)-mediated fibrogenic pathways. Much of our knowledge of CCN2 in fibrosis has come from studies in which its production or activity have been experimentally attenuated. These studies, performed both in vitro and in animal models, have demonstrated the utility of pharmacological inhibitors (e.g. tumor necrosis factor alpha (TNF alpha), prostaglandins, peroxisome proliferator-activated receptor-gamma (PPAR gamma) agonists, statins, kinase inhibitors), neutralizing antibodies, antisense oligonucleotides, or small interfering RNA (siRNA) to probe the role of CCN2 in fibrogenic pathways. These investigations have allowed the mechanisms regulating CCN2 production to be more clearly defined, have shown that CCN2 is a rational anti-fibrotic target, and have established a framework for developing effective modalities of therapeutic intervention in vivo. PMID- 19294532 TI - Protective effect of betaine on changes in the levels of lysosomal enzyme activities in heart tissue in isoprenaline-induced myocardial infarction in Wistar rats. AB - Myocardial infarction is one of the most common manifestations of cardiovascular disease. In the present study, we investigated the protective effect of betaine, a potent lipotropic molecule, on changes in the levels of lysosomal enzymes and lipid peroxidation in isoprenaline-induced myocardial infarction in Wistar rats, an animal model of myocardial infarction in man. Male albino Wistar rats were pretreated with betaine (250 mg/kg body weight) daily for a period of 30 days. After the treatment period, isoprenaline (11 mg/100 g body weight) was intraperitoneally administered to rats at intervals of 24 h for 2 days. The activities of lysosomal enzymes (beta-glucuronidase, beta-galactosidase, beta glucosidase, and acid phosphatase) were significantly (p < 0.05) increased in plasma with a concomitant decline in the activities of these enzymes in heart tissue of isoprenaline-administered rats. Also, the level of lipid peroxidation was higher in heart lysosomes of isoprenaline-injected rats. Pretreatment with betaine daily for a period of 30 days to isoprenaline-induced rats prevented the changes in the activities of these lysosomal enzymes. Oral treatment with betaine (250 mg/kg body weight) to normal control rats did not show any significant effect in all the biochemical parameters studied. Thus, the results of our study show that betaine protects the lysosomal membrane against isoprenaline-induced myocardial infarction. The observed effects might be due to the free radical scavenging and membrane-stabilizing properties of betaine. PMID- 19294533 TI - Prognostic value of tumor-related molecular expression in gastric carcinoma. AB - In order to identify reliable molecular markers for prognostic prediction in gastric carcinoma, we evaluated the expression of six molecular markers, namely bFGF, IGF-2, HGF, MMP-9, integrin beta3 and uPA in gastric cancer. There was a significant correlation between the expression of these markers and the depth of tumor invasion, vessel invasion, lymph node and distant metastasis, TNM stage and microvessel density. The average survival time and 5-year survival rate of patients with positive expression of molecular markers was higher than those with negative expression. Multivariate analysis showed that abnormal expression of bFGF, MMP-9 and uPA, as well as depth of invasion, lymph node and distant metastasis and TNM stage were independently related to poor prognosis of gastric cancer. MMP-9, bFGF and uPA are potential candidates for development as clinically applicable molecular prognostic markers for gastric carcinoma, and may be effective therapeutic targets for the disease in the future. PMID- 19294534 TI - Antioxidants but not doxycycline treatments restore depressed beta-adrenergic responses of the heart in diabetic rats. AB - Reactive oxygen species (ROS) play important roles in the development of diabetic cardiomyopathy. Matrix metalloproteinases (MMPs) can get activated by ROS and contribute to loss of myocardial contractile function in oxidative stress injury. Previously we have shown that either a MMP-2 inhibitor doxycycline or an antioxidant selenium treatment in vivo prevented diabetes-induced cardiac dysfunction significantly. In addition, there is an evidence for impaired cardiac responsiveness to beta-adrenoceptor (beta AR) stimulation in experimental animals with diabetes. The exact nature of linkage between the functional depression in cardiac responses to catecholamines and the variations in uncoupling of beta AR in diabetes has not been clearly defined. Therefore, we aimed to evaluate the effect of in vivo administration of doxycycline on beta AR responses of isolated hearts from diabetic rats and compare these data with two well-known antioxidants; sodium selenate and (n-3) fatty acid-treated diabetic rats. We examined the changes in the basal cardiac function in response to the beta AR stimulation, adenylate cyclase activity, and beta AR affinity to its agonist, isoproterenol. These results showed that antioxidant treatment of diabetic rats could protect the hearts against diabetes-induced depression in beta AR responses, significantly while doxycycline did not have any significant beneficial action on these parameters. As a summary, present data, in part, demonstrate that antioxidants and MMP inhibitors could both regulate MMP function but may also utilize different mechanisms of action in cardiomyocytes, particularly related with beta AR signaling pathway. PMID- 19294535 TI - Intra-arterial air thrombogenesis after cerebral air embolism complicating lower extremity sclerotherapy. AB - BACKGROUND: Cerebral arterial gas embolism is a potentially life-threatening event. Intraarterial air can occlude blood flow directly or cause thrombosis. Sclerotherapy is an extremely rare cause of cerebral arterial gas embolism. METHOD: Case-report RESULTS: A 38-year-old female suffered acute onset of a left middle cerebral artery (LMCA) syndrome with an NIH stroke score of 11 approximately 10 min after lower extremity sclerotherapy. CT angiogram demonstrated LMCA intraarterial air. Patient fully recovered after hyperbaric oxygen treatment with complete resolution of intraarterial air. However, thrombus replaced intraarterial air despite anticoagulation with heparin. CONCLUSION: We provide radiological evidence of hyperbaric oxygen therapy resolving intraarterial air but also demonstrate the thrombogenic potential of this procedural complication. PMID- 19294536 TI - Stage-specific survival differences associated with postoperative radiotherapy for gastrointestinal cancers. AB - OBJECTIVE: To study the frequency and effect associated with postoperative radiotherapy (RT) for patients with resected gastrointestinal (GI) cancers. MATERIALS AND METHODS: In observational cohort from the Surveillance, Epidemiology, and End Results (SEER) program, a total of 23,049 patients were identified with resected pancreatic, gastric, esophageal, or rectal carcinomas diagnosed from 1988 to 2003. Using a propensity score analysis, survival differences associated with postoperative RT were analyzed. RESULTS: Adjuvant RT was given to 51.2%, 26.3%, 33.0%, and 58.0% of pancreatic, gastric, esophageal, and rectal cancer patients, respectively. Age and stage of disease were associated with RT use for each site (P < 0.001), with younger patients and those with advanced disease receiving RT more frequently. Postoperative RT was associated with a survival benefit for patients with pancreatic cancer (hazard ratio [HR], 0.87; 95% confidence interval [CI], 0.79-0.96), gastric cancer (HR, 0.93; 95% CI, 0.87-0.99), and rectal cancer (HR, 0.84; 95% CI, 0.79-0.90). Subgroups of patients were also identified who experienced the greatest improvement in survival with RT (stage IIB pancreatic cancer, HR = 0.71 [95% CI 0.62-0.80]; stage IIIA and IV gastric cancer, HR = 0.86 [95% CI 0.77-0.97] and HR = 0.77 [95% CI 0.67-0.89], respectively; stages IIA, IIIB, and IIIC rectal cancer, HR = 0.87 [95% CI 0.78-0.97], HR = 0.71 [95% CI 0.63-0.80], and HR = 0.79 [95% CI 0.70-0.90], respectively). CONCLUSION: Postoperative RT is associated with improved survival for patients who undergo curative resection of pancreatic, gastric, and rectal malignancies. Significant differences are observed for this effect according to stage of disease, with more advanced cases in general experiencing a greater benefit with RT. PMID- 19294537 TI - Stenosis of gastric body as a rare complication after endoscopic submucosal dissection for multiple gastric epithelial tumors. AB - We experienced a rare case of stenosis of the gastric body, a wider tract, that occurred after multiple, wide endoscopic resection five times for four lesions of early gastric cancer and gastric adenomas. We report this case as a sentinel experience of endoscopic submucosal dissection of the stomach. PMID- 19294538 TI - Analysis of a case with disappearance of the primary gastrointestinal stromal tumor and progressive liver metastases under long-term treatment with tyrosine kinase inhibitors. AB - The response of gastrointestinal stromal tumors (GISTs) to tyrosine kinase receptor inhibitors (TKR-I) has been a breakthrough for small molecular therapy. We report here on the very different long-term outcome of a synchronous metastatic GIST with complete remission of the primary tumor and progressive liver metastases under TKR-I therapy. In 2003, a 52-year-old patient was diagnosed with gastric GIST and synchronous multiple liver metastases. Therapy with imatinib, 400 mg daily, was started immediately. Fifteen months later, the primary was no longer detectable by endoscopy. In 2006, progression of the liver metastases was observed. Mutation analysis of the initial biopsy specimen from the primary, as well as the biopsy from the three main liver metastases after 3 years of imatinib treatment, revealed the common KIT exon 11 deletion (W557_K558del) in all tumor samples. Two of the metastases had a separate secondary mutation in KIT exon 14 and 17, respectively, while the largest cystic metastatic lesion had no other mutation. Imatinib was then increased to a daily dose of 800 mg, and in April 2007 the treatment was changed to sunitinib. Fifty two months after initial diagnosis, the patient died of liver failure. At no time point, relapse of the gastric primary tumor was observed. Whilst TKR-Is are commonly very effective in treating GISTs, the present case illustrates their varying effects regarding the clinical behavior and genetic variations within different tumors of the same patient after long-term treatment. PMID- 19294539 TI - Fatal retropharyngeal abscess: a possible marker of inflicted injury in infancy and early childhood. AB - Retropharyngeal abscesses are serious infections of the deep tissues of the neck associated with significant morbidity and mortality due to their proximity to vital structures. The most common age range is 2-4 years, with neonates accounting for approximately 10% of cases. The following report demonstrates possible forensic issues that may arise in such cases. CASE REPORT: A 10-week-old infant was found dead in her father's arms an hour after feeding. At autopsy death was due to a large retropharyngeal abscess with disseminated Streptococcus pneumoniae sepsis. Other findings at autopsy of bruising and multiple fractures of the ribs and limbs indicated inflicted injury; this raised the possibility that the abscess had arisen from forceful foreign body insertion into the pharynx. The mortality and complication rates of retropharyngeal abscess remain high even with the institution of appropriate treatment; however, the wide variety of presentations often causes delays in diagnosis and treatment. When found at autopsy in infants and children the possibility of inflicted injury should be considered. PMID- 19294540 TI - Follow up your unexpected clinical observations! PMID- 19294541 TI - Utilisation of specialist care in patients with incurable rectal cancer. a population-based study from Western Norway. AB - INTRODUCTION: About 25% of patients with rectal cancer have incurable disease at the time of diagnosis. In the current study from Western Norway (population of 981 000) we focused on the utilisation of specialist care in patients with primarily incurable rectal cancer. PATIENTS AND METHODS: Between 1997 and 2002, 1 167 patients were diagnosed with rectal cancer, of whom 297 (25%) had incurable disease, according to consecutive and prospective reporting to the Norwegian Colorectal Cancer Registry. Consumption of specialist care facilities was studied with regard to outpatient contacts, hospital admissions, and various treatment modalities. Data were analysed with regard to age, sex, marital status, type of residence, and geographical access to hospital facilities. Data were available for 287 patients (97%). RESULTS: The median age was 77 years. Elderly patients (>77 years) more often lived in nursing homes without a spouse. About 60% of the patients were treated with major surgery, chemotherapy or radiotherapy, either alone or in combination. Of those who did not receive such treatment, 87% were elderly. Oncological treatment, either alone or combined with surgery, predicted increased hospital admissions and outpatient contacts. Age >77 years predicted fewer hospital admissions. Survival varied statistically significantly with the various treatment modalities, and was highest for major resections combined with oncological treatment. The majority of the patients living at home died in hospitals (54%) and only 26% died in their homes, while two-thirds of residents of nursing homes died there. DISCUSSION: Patients with primary incurable rectal cancer are heterogeneous with regard to their needs of treatment. While younger patients receive extensive tumour-related treatment, elderly patients are most commonly treated according to their symptoms. Prospective studies of the effect of various treatment options on the ease of symptoms and improved quality of life in unselected populations are needed. PMID- 19294542 TI - Dynamic response of wall shear stress on the stenosed artery. AB - The present study deals with an appropriate mathematical model of an artery in the presence of constriction in which the generated wall shear stress due to blood flow is analysed. The geometry of the stenosed arterial segment in the diseased state, causing malfunction of the cardiovascular system, is formed mathematically. The flowing blood contained in the stenosed artery is treated as non-Newtonian and the flow is considered to be two-dimensional. The motion of the arterial wall and its effect on local fluid mechanics is not ruled out from the present pursuit. The flow analysis applies the time-dependent, two-dimensional incompressible nonlinear Navier-Stokes equations for non-Newtonian fluids. The flow-field can be obtained primarily following the radial coordinate transformation, using the appropriate boundary conditions and finally adopting a suitable finite difference scheme numerically. The influences of flow unsteadiness, the arterial wall distensibility and the presence of stenosis on the flow-field and the wall shear stresses are quantified in order to indicate the susceptibility to atherosclerotic lesions and thereby to validate the applicability of the present theoretical model. PMID- 19294543 TI - Simulation of biological growth. AB - Researchers concerned with the growth of biological tissue often use models that predict the growth as a function of a mechanical stimulus such as stress, strain or elastic energy. However, a general theory for bulk growth should consider that the mechanical stimulus may only be one of many factors contributing to growth. Another important factor could be time, as living tissues can be assumed to have a pre-programmed directional biological growth that is independent of mechanical stimuli. This paper has two objectives: the first is to introduce the concept of directional biological growth within a well developed growth theory, the second is to present the computational methods by which three-dimensional growth that encompasses time and stress effects can be simulated using commercially available finite element analysis software. PMID- 19294544 TI - SWOT analysis of a pediatric rehabilitation programme: a participatory evaluation fostering quality improvement. AB - PURPOSE: To present the results of a strengths, weaknesses, opportunities and threats (SWOT) analysis used as part of a process aimed at reorganising services provided within a pediatric rehabilitation programme (PRP) in Quebec, Canada and to report the perceptions of the planning committee members regarding the usefulness of the SWOT in this process. METHOD: Thirty-six service providers working in the PRP completed a SWOT questionnaire and reported what they felt worked and what did not work in the existing model of care. Their responses were used by a planning committee over a 12-month period to assist in the development of a new service delivery model. Committee members shared their thoughts about the usefulness of the SWOT. RESULTS: Current programme strengths included favourable organisational climate and interdisciplinary work whereas weaknesses included lack of psychosocial support to families and long waiting times for children. Opportunities included working with community partners, whereas fear of losing professional autonomy with the new service model was a threat. The SWOT results helped the planning committee redefine the programme goals and make decisions to improve service coordination. SWOT analysis was deemed as a very useful tool to help guide service reorganisation. CONCLUSIONS: SWOT analysis appears to be an interesting evaluation tool to promote awareness among service providers regarding the current functioning of a rehabilitation programme. It fosters their active participation in the reorganisation of a new service delivery model for pediatric rehabilitation. PMID- 19294545 TI - What factors predict full or partial return to work among sickness absentees with spinal pain participating in rehabilitation? AB - PURPOSE: To identify the factors that predict full or partial return to work among long-term (> or =90 days) sickness absentees due to spinal pain who begin a multidisciplinary rehabilitation programme. METHOD: In a prospective cohort study, 312 patients with neck, thoracic and/or lumbar pain, aged 20-64, participated in a 4-week multidisciplinary rehabilitation programme in Sweden. Questionnaire data at inclusion were used. Factors included in logistic regressions were as follows: age, gender, type of work, pain location, pain intensity (visual analogue scale), activity limitations [Disability Rating Index (DRI)], health-related quality of life (SF-36), pain-related fear of movement (Tampa Scale of Kinesiophobia), motivation (Self Motivation Inventory), sickness absence at baseline and number of sick-leave days during the previous 2 years. Outcome factor was increased versus not increased working time at follow-up 6 months later. RESULTS: Most patients (68%) reported two or three pain locations. At baseline, 56% were full-time sickness absent and 23% at follow-up; 61% had increased their working time. Predictors for increased working time were age below 40 years, low activity limitation (DRI < 50), low SF-36 bodily pain (> 30) and high SF-36 social functioning (> 60). Number of sick-leave days during the previous 2 years (md 360; range 90-730) had no influence. CONCLUSIONS: Even patients with long previous sick leave can increase working time after a multidisciplinary rehabilitation programme, especially if they are younger, have lower levels of activity limitations and pain and better social functioning. To include information on part-time work is useful when evaluating work ability following rehabilitation programmes. PMID- 19294546 TI - Predictors of daily mobility skills 6 months post-discharge from acute care or rehabilitation in older adults with stroke living at home. AB - PURPOSE: To determine the evolution of daily mobility skills from the timed up and-go (TUG) upto 6 months after home return in older adults with stroke discharged from acute care or rehabilitation; and to identify the best predictive factors of the TUG at 6 months post-discharge. METHODS: In this longitudinal prospective study, people with stroke aged 65 years or more and discharged home from an acute care hospital (n = 82) or a rehabilitation service (n = 109) were included. The TUG was measured at discharge (T1), and at 3 and 6 months post discharge (T2 and T3). Correlations between the TUG at T3 and sociodemographic and clinical variables, as well as physical, cognitive, perceptual and psychological measures at T1, were used in a multiple regression model to identify the best predictors of TUG at T3. RESULTS: TUG did not change between T1, T2 and T3 in the two groups of participants. The best predictors of TUG at T3 in participants from acute care were the use of a walking aid in daily life, age, deficits in oral expression and the presence of depressive symptoms. In participants from rehabilitation, predictors were the stage of motor recovery of the foot, the use of a walking aid in daily life, number of schooling years and memory impairments. CONCLUSION: Daily mobility skills, as assessed with the TUG, did not deteriorate upto 6 months after home return in older adults with stroke. The best predictor of the TUG at T3 is the use of a walking aid during daily life in participants from acute care, and motor recovery of the foot in participants from rehabilitation. PMID- 19294547 TI - What type of service provision do patients with chronic pain want from primary care providers? AB - PURPOSE: The aim of this study was to explore what types of service provision patients with chronic pain wanted from their general practitioners (GP). METHOD: A small scale survey measured anxiety and depression and quantified the extent to which patients wanted four different types of help from their GP (explanation and understanding, medical treatment, psychological support and information). An opportunistic sample of 155 patients (30.3% male and 69.7% female) was recruited from three general practice surgeries in the northwest region of Northern Ireland. Ninety-one participants self-reported chronic pain and there were 64 in the no pain group. RESULTS: Even after statistically controlling for anxiety and depression, individuals in the chronic pain group had a greater need for emotional/psychological support and explanation and understanding from their GPs. There were no significant differences between the groups with respect to the need for more information or medical treatment. CONCLUSIONS: A biopsychosocial approach should be employed in the management of chronic pain, however as this study shows, some primary care patients are still being treated within a biomedical framework. To address the unmet needs of this patient population, there is a need for clinical guidelines in the management of chronic pain in primary care settings. Limitations of the present research, with recommendations for future study are offered. PMID- 19294548 TI - Effect of chitosan glutamate, carbomer 974P, and EDTA on the in vitro Caco-2 permeability and oral pharmacokinetic profile of acyclovir in rats. AB - BACKGROUND: Chitosan glutamate and polyacrylic acid (e.g., carbomer 974P) are known to modulate the tight junctions in the intestinal wall and increase permeability and blood exposure of drugs absorbed orally by the paracellular route. AIM: To assess the impact of chitosan glutamate and carbomer 974P on the absorption of paracellularly absorbed model drug, acyclovir, in vitro and in rat in vivo. METHODS: The influence of chitosan glutamate and carbomer 974P (alone and in combination with EDTA-Na2) on the in vitro Caco-2 permeability and oral pharmacokinetic profile in the rat of acyclovir was investigated. RESULTS: In the presence of chitosan glutamate, the apparent permeability of acyclovir across Caco2 monolayer increased 4.1 times relative to control. This increase was accompanied by a significant ( approximately 60%) decrease in transepithelial electrical resistance values indicating opening of the tight junctions in the cell monolayer. In rat, chitosan glutamate doubled oral bioavailability of acyclovir and tripled the amount of acyclovir excreted unchanged into urine. In contrast, the effect of carbomer 974P was not statistically significant at 5% level. CONCLUSIONS: In conclusion, chitosan glutamate (1-3%) and chitosan glutamate (1%)/EDTA-Na2 (0.01%) are effective excipients to increase permeability of acyclovir across Caco-2 cell monolayers and the oral absorption in the rat in vivo. PMID- 19294549 TI - Stem cells: comprehensive treatments for amyotrophic lateral sclerosis in conjunction with growth factor delivery. AB - Amyotrophic lateral sclerosis (ALS) is characterized by loss of both upper and lower motor neurons. ALS progression is complex and likely due to cellular dysfunction at multiple levels, including mitochondrial dysfunction, glutamate excitotoxicity, oxidative stress, axonal dysfunction, reactive astrocytosis, and mutant superoxide dismutase expression, therefore, treatment must provide neuronal protection from multiple insults. A significant amount of ALS research focuses on growth factor-based therapies. Growth factors including insulin-like growth factor-I, vascular endothelial growth factor, brain-derived neurotrophic factor, and glial-derived neurotrophic factor exhibit robust neuroprotective effects on motor neurons in ALS models. Issues concerning growth factor delivery, stability and unwanted side effects slow the transfer of these treatments to human ALS patients. Stem cells represent a new therapeutic approach offering both cellular replacement and trophic support for the existing population. Combination therapy consisting of stem cells expressing beneficial growth factors may provide a comprehensive treatment for ALS. PMID- 19294550 TI - Comparisons of stuttering frequency during and after speech initiation in unaltered feedback, altered auditory feedback and choral speech conditions. AB - BACKGROUND: Stuttering is prone to strike during speech initiation more so than at any other point in an utterance. The use of auditory feedback (AAF) has been found to produce robust decreases in the stuttering frequency by creating an electronic rendition of choral speech (i.e., speaking in unison). However, AAF requires users to self-initiate speech before it can go into effect and, therefore, it might not be as helpful as true choral speech during speech initiation. AIMS: To examine how AAF and choral speech differentially enhance fluency during speech initiation and in subsequent portions of utterances. METHODS & PROCEDURES: Ten participants who stuttered read passages without altered feedback (NAF), under four AAF conditions and under a true choral speech condition. Each condition was blocked into ten 10 s trials separated by 5 s intervals so each trial required 'cold' speech initiation. In the first analysis, comparisons of stuttering frequencies were made across conditions. A second, finer grain analysis involved examining stuttering frequencies on the initial syllable, the subsequent four syllables produced and the five syllables produced immediately after the midpoint of each trial. OUTCOMES & RESULTS: On average, AAF reduced stuttering by approximately 68% relative to the NAF condition. Stuttering frequencies on the initial syllables were considerably higher than on the other syllables analysed (0.45 and 0.34 for NAF and AAF conditions, respectively). After the first syllable was produced, stuttering frequencies dropped precipitously and remained stable. However, this drop in stuttering frequency was significantly greater (approximately 84%) in the AAF conditions than in the NAF condition (approximately 66%) with frequencies on the last nine syllables analysed averaging 0.15 and 0.05 for NAF and AAF conditions, respectively. In the true choral speech condition, stuttering was virtually (approximately 98%) eliminated across all utterances and all syllable positions. CONCLUSIONS & IMPLICATIONS: Altered auditory feedback effectively inhibits stuttering immediately after speech has been initiated. However, unlike a true choral signal, which is exogenously initiated and offers the most complete fluency enhancement, AAF requires speech to be initiated by the user and 'fed back' before it can directly inhibit stuttering. It is suggested that AAF can be a viable clinical option for those who stutter and should often be used in combination with therapeutic techniques, particularly those that aid speech initiation. The substantially higher rate of stuttering occurring on initiation supports a hypothesis that overt stuttering events help 'release' and 'inhibit' central stuttering blocks. This perspective is examined in the context of internal models and mirror neurons. PMID- 19294551 TI - Older people who stutter: barriers to communication and perceptions of treatment needs. AB - BACKGROUND: Little is known about the experience of stuttering for people over 55 years of age. Recent research has established that the same types of stuttering behaviours, cognitions, and emotional consequences experienced during young adulthood persist into older age. AIMS: The aims were to investigate perceptions of limitations to activity and participation in a group of older people who stuttered into adulthood. A further aim was to find out their perceptions about treatment. METHODS & PROCEDURES: This was a qualitative study involving eleven participants, eight males and three females over 55 years of age (mean age = 70.7, standard deviation = 9.13 years, range = 57.2-83.8 years) who self-reported stuttering into adulthood. Participants were randomly assigned to two focus groups for the discussion of topic questions posed by a moderator. The discussion was video- and audio-recorded, transcribed, and analysed using a comparative thematic analysis to derive emergent themes in relation to the topic questions. OUTCOMES & RESULTS: Stuttering can impact on the lives of older people in a similar way to younger people who stutter. Participants who continued to work felt more limited by their stuttering because work involved unpredictable speaking situations with unfamiliar people. Others who had retired experienced some relief from these limitations because they were no longer required to communicate in a work context. The acceptance of stuttering was a theme expressed by some participants, and acceptance diminished the limitations because these older people were less fearful of the consequences of their stuttering. However, others remained constrained by the impact of stuttering on their communication and struggled with a fear of speaking and a fear of negative evaluation by others. They applied learnt and self-devised techniques to assist their speech and felt that if fear of speaking was removed and their self-confidence increased, communication might be better. They would like effective, individual, and short-term treatment with speech-language pathologists who are knowledgeable about stuttering and sensitive to their emotional needs. CONCLUSIONS & IMPLICATIONS: Older people who stutter experienced limitations to participation because of their stuttering and there are implications for their future ability to remain independent and connected to relevant people and services. Further investigation of these limitations and research into effective intervention is indicated. PMID- 19294552 TI - Why do speech and language therapists stay in, leave and (sometimes) return to the National Health Service (NHS)? AB - BACKGROUND: Research into recruitment, retention and return of speech and language therapists in the National Health Service (NHS) is relatively limited, particularly in respect of understanding the factors that drive employment choice decisions. AIMS: To identify what factors influence speech and language therapists working in the NHS to stay, and consider leaving, but not do so. To identify what features of the NHS and alternative employers influence speech and language therapists to leave the NHS. To identify why some speech and language therapists return to the NHS after working elsewhere. METHODS & PROCEDURES: A total of 516 male and female speech and language therapists, in three distinct groups (NHS stayers, leavers and returners) completed a questionnaire and gave responses to open-ended questions regarding their perceptions of the NHS and other employers. Qualitative data analysis identified reasons why individuals stayed in, left or returned to the NHS employment, and what actions could be taken by management to facilitate retention and return. OUTCOMES & RESULTS: Stayers value job and pension security; professional development opportunities; the work itself; and professional support. Leavers not involved in childcare left because of workload/pressure/stress; poor pay; and not being able to give good patient care. Returners returned because of flexible hours; work location; professional development; and pension provision. Stayers and returners primarily wish to see more staff in the NHS, whereas leavers would return if there were more flexibility in work arrangements. Returners were particularly hostile towards Agenda for Change. CONCLUSIONS & IMPLICATIONS: Whilst some preferences appear to require increased resources, others such as reducing bureaucracy and valuing professionals do not. The full impact of Agenda for Change has yet to be established. Predicted excess labour supply of allied health professionals and future structural changes present pressures but also possible opportunities for speech and language therapists. PMID- 19294553 TI - Cognitive-linguistic deficit and speech intelligibility in chronic progressive multiple sclerosis. AB - BACKGROUND: Multiple sclerosis is a disabling neurological disease with varied symptoms, including dysarthria and cognitive and linguistic impairments. Association between dysarthria and cognitive-linguistic deficit has not been explored in clinical multiple sclerosis studies. AIMS: In patients with chronic progressive multiple sclerosis, the study aimed to investigate the presence and nature of cognitive-linguistic deficit, the association between levels of cognitive-linguistic ability and speech intelligibility, and of both of these with functional disability and time since onset of multiple sclerosis symptoms. METHODS & PROCEDURES: The Arizona Battery for Communication Disorders of Dementia (ABCD) (Bayles and Tomoeda 1993 ), The Assessment of Intelligibility of Dysarthric Speech (AIDS) Sentence Intelligibility Task (Yorkston and Beukelman 1984 ), and the Modified Barthel Activities of Daily Living Index (MBADLI) (Shah 1998 ) were administered to 24 chronic progressive multiple sclerosis participants with dysarthria. A total of 24 non-neurologically impaired participants, matched for gender, age and education, formed a control group. OUTCOMES & RESULTS: For multiple sclerosis participants, linear regression analysis showed a strong association between ABCD and AIDS (beta = 0.89, p = 0.005), no association between ABCD and either MBADLI or time since onset, a strong association between AIDS and MBADLI (beta = 0.60, p = 0.001), and a trend towards an association between AIDS and time since onset (beta = -0.29, p = 0.08). Correlations between the four included ABCD construct scores and between these and the total ABCD score were significant (r>0.60, p<0.01). For each of the 15 included ABCD measures and for the four construct scores and the overall ABCD score, multiple sclerosis and control group performances were significantly different (p<0.01) and effect sizes were large (d>0.80). CONCLUSIONS & IMPLICATIONS: The results revealed a strong association between dysarthria, as measured by connected speech intelligibility testing, and cognitive-linguistic deficit, in people with chronic progressive-type multiple sclerosis. While some of the impairments that are associated with multiple sclerosis, including motor speech disorder, may influence performance on the ABCD, the data support the conclusion that marked cognitive-linguistic deficit is present in chronic progressive-type multiple sclerosis patients with dysarthria. Deterioration was global, rather than being indicative of a construct specific deficit, and encompassed language, both expression and comprehension. Episodic memory and linguistic expression were especially affected. Speech and language therapists who work with dysarthric patients with chronic progressive multiple sclerosis should monitor cognitive-linguistic impairment. An awareness of this might influence assessment, intervention and management, including the information and advice given to patients and their relatives. PMID- 19294554 TI - Improved vocabulary production after naming therapy in aphasia: can gains in picture naming generalize to connected speech? AB - BACKGROUND: Naming accuracy for nouns and verbs in aphasia can vary across different elicitation contexts, for example, simple picture naming, composite picture description, narratives, and conversation. For some people with aphasia, naming may be more accurate to simple pictures as opposed to naming in spontaneous, connected speech; for others, the opposite pattern may be evident. These differences have, in some instances, been related to word class (for example, noun or verb) as well as aphasia subtype. Given that the aim of picture naming therapies is to improve word-finding in general, these differences in naming accuracy across contexts may have important implications for the potential functional benefits of picture-naming therapies. AIMS: This study aimed to explore single-word therapy for both nouns and verbs, and to answer the following questions. (1) To what extent does an increase in naming accuracy after picture naming therapy (for both nouns and verbs) predict accurate naming of the same items in less constrained spontaneous connected speech tasks such as composite picture description and retelling of a narrative? (2) Does the word class targeted in therapy (verb or noun) dictate whether there is 'carry-over' of the therapy item to connected speech tasks? (3) Does the speed at which the picture is named after therapy predict whether it will also be used appropriately in connected speech tasks? METHODS & PROCEDURES: Seven participants with aphasia of varying degrees of severity and subtype took part in ten therapy sessions over five weeks. A set of potentially useful items was collected from control participant accounts of the Cookie Theft Picture Description and the Cinderella Story from the Quantitative Production Analysis. Twenty-four of these words (twelve verbs and twelve nouns) were collated for each participant, on the basis that they had failed to name them in either simple picture naming or connected speech tasks (picture-supported narrative and unsupported retelling of a narrative). These were placed in a larger cohort of verb and noun sets for therapy. Post-therapy assessments examined naming accuracy and speed of target items in single-word picture-naming and naming accuracy in connected speech contexts. OUTCOMES & RESULTS: There was a step-wise decrement in naming accuracy over the three naming contexts following targeted therapy. Simple pictures elicited the most correct names, followed by picture-supported narratives and lastly unsupported narratives. Picture-naming accuracy significantly predicted naming in the connected speech contexts for the group as a whole. The speed of picture naming after therapy did not predict the extent to which items were named in composite picture description and narrative tasks. CONCLUSIONS & IMPLICATIONS: The findings suggest that gains in naming accuracy obtained through picture naming therapy may generalize to naming of the same items in more linguistically and cognitively demanding connected speech tasks. Demonstrating this generalization is methodologically challenging and the method utilized in this study may serve as one starting point for gathering a larger database in order to answer the question posed by this paper more robustly. PMID- 19294555 TI - The barriers perceived to prevent the successful implementation of evidence-based practice by speech and language therapists. AB - BACKGROUND: There is currently a paucity of research investigating what speech and language therapists, in particular, perceive are the greatest barriers to implementing evidence-based practice. AIMS: The purpose of this study was to investigate the perceived barriers that are faced by speech and language therapists in southern Ireland when attempting to implement evidence-based practice. METHODS & PROCEDURES: A 34-item questionnaire was sent to 39 therapists working in several counties in southern Ireland. The survey received an 82.1% (n = 32) response rate. OUTCOMES & RESULTS: The results of the study indicated that certain barriers are perceived to prevent evidence-based practice being implemented successfully. The most significant barrier affecting evidence-based practice implementation was reported to be a lack of time to read research (71.9%). Additional barriers that were found to be the most significant were the research having methodological inadequacies (62.5%) and insufficient time to implement new ideas (59.4%). Other important factors identified as being significant barriers to the implementation of evidence-based practice were those associated with the quality and presentation of the research, workplace setting, and lack of skills of the therapist. Associations between specific barriers and workplace setting or grade were also investigated. Some possible reasons for these barriers and the implications for clinical practice are also discussed. CONCLUSIONS & IMPLICATIONS: This small study suggests that therapists agreed that evidence-based practice is essential to the practice of speech and language therapy. There are, however, barriers in place that are perceived to prevent its successful implementation. It is hoped that because these barriers have been identified, individual clinicians and organizations can be proactive in aiming to provide an evidence-based service to their clients. PMID- 19294556 TI - Diagnostic delay and complications for older adults with multiple myeloma. AB - Increased attention to timely diagnosis motivated us to study 5483 patients diagnosed with multiple myeloma using Medicare claims linked to tumor registries in the Surveillance, Epidemiology and End Results programme. We calculated the time between initial visits for anemia or back pain and for myeloma diagnosis, and used logistic regression to predict the likelihood of diagnostic delay, and also the likelihood of renal or skeletal complications. The median time between sign or symptom and myeloma diagnosis was 99 days. Patients with anemia, back pain and comorbidities were more likely to experience diagnostic delay (OR 1.6, 95% CI 1.3-2.0). Diagnosis while hospitalised (OR 2.5, 95% CI 2.2-2.9) and chemotherapy treatment within 6 months of diagnosis (OR 1.4, 95% CI 1.2-1.6) significantly predicted complications; diagnostic delay did not (OR 0.9, 95% CI 0.8-1.1). Our data suggest that complications are more strongly associated with health status and myeloma severity than with diagnostic delays. PMID- 19294557 TI - Evaluation of a low density DNA microarray for small B-cell non-Hodgkin lymphoma differential diagnosis. AB - Lymphomas are classified according to the World Health Organisation (WHO) classification which defines subtypes on the basis of clinical, morphological, immunophenotypic, molecular and cytogenetic criteria. Differential diagnosis of the subtypes is sometimes difficult, especially for small B-cell lymphoma (SBCL). Standardisation of molecular genetic assays using multiple gene expression analysis by microarrays could be a useful complement to the current diagnosis. The aim of the present study was to develop a low density DNA microarray for the analysis of 107 genes associated with B-cell non-Hodgkin lymphoma and to evaluate its performance in the diagnosis of SBCL. A predictive tool based on Fisher discriminant analysis using a training set of 40 patients including four different subtypes (follicular lymphoma n = 15, mantle cell lymphoma n = 7, B cell chronic lymphocytic leukemia n = 6 and splenic marginal zone lymphoma n = 12) was designed. A short additional preliminary analysis to gauge the accuracy of this signature was then performed on an external set of nine patients. Using this model, eight of nine of those samples were classified successfully. This pilot study demonstrates that such a microarray tool may be a promising diagnostic approach for small B-cell non-Hodgkin lymphoma. PMID- 19294558 TI - T-Cell acute lymphoblastic leukemia with a "pinch" of BCR-ABL1. PMID- 19294559 TI - Distinct neutrophil subpopulations phenotype by flow cytometry in myelodysplastic syndromes. AB - The cardinal feature of myelodysplastic syndromes (MDS) is dysplasia involving one or more myeloid cell lineages. In the present study, we used 4-color flow cytometric analysis to investigate dysgranulopoiesis in bone marrow specimens from 65 patients with MDS. The antigen expression patterns of total neutrophil granulocytes (TNG) and of the two distinct neutrophil granulocytic subpopulations (NGSs), NGS-1 (dimmer CD45 expression) and NGS-2 (stronger CD45 expression) identified on the side scatter (SS) vs. CD45-intensity plot, were studied. The neutrophil granulocytes from patients with MDS showed characteristic antigen expression aberrancies which were more pronounced in NGS-2 subpopulation. Studying separately the NGS-2 subpopulation with the CD16/MPO/LF combination, the low CD16(+)/MPO(+) and low CD16(+)/LF(+) percentages seemed to discriminate between lower-risk and higher-risk patients with MDS in most occasions. Furthermore, a detailed assessment of the NGS-1 and NGS-2 immunophenotypic patterns revealed early dysplastic changes, not otherwise observed by standard TNG analysis, especially in cases of lower-risk MDS. PMID- 19294560 TI - Clinical management of massive retroperitoneal hemorrhage after bone marrow biopsy. PMID- 19294561 TI - Liposomal cytarabine in advanced-stage acute lymphoblastic leukemia and aggressive lymphoma with central nervous system involvement: experience of the Polish Acute Leukemia Group. PMID- 19294562 TI - Collaborative memory intervention in dementia: caregiver participation matters. AB - The aim of the study was to examine the effectiveness of a collaborative memory intervention for persons with dementia and their spousal caregivers, where the couple acquired and practised memory supportive strategies (spaced-retrieval and hierarchical cuing) to learn a face-name association and to set a table for coffee/tea. The collaborative intervention was compared to an individual intervention where the person with dementia received the same training but without the participating caregiver and to a control group of couples receiving no training. The results showed that following collaborative intervention recall performance in two collaborative tasks became more equally shared between the spouses, reflected in a decrease in recall for the caregiver and in an increase in recall for the spouse with dementia between pre- and post-test; whereas for the other two groups the caregivers dominated collaborative recall both at pre- and post-test. The results also showed that the persons with dementia in the collaborative group improved their individually assessed episodic memory performance as compared to the persons with dementia in the other two groups. Finally, training had no effects on episodic memory, perceived burden or depressive symptoms for the caregivers. These results suggest that the active participation of the caregiver matters in cognitive dementia rehabilitation. PMID- 19294564 TI - Ocular Immunology and Inflammation. Editorial. PMID- 19294565 TI - Controversies in intraocular lymphoma. PMID- 19294566 TI - Intraocular lymphoma: more questions than answers. PMID- 19294567 TI - Causes of uveitis at a referral center in Saudi Arabia. AB - PURPOSE: The main purpose of this study was to determine the causes of uveitis at the Eye Center in Saudi Arabia. METHODS: A retrospective review. The causes of uveitis were analyzed according to various patient characteristics. RESULTS: A total of 488 cases were encountered. The age range was 3 to 99 years (mean age: 38 years). The most common causes of uveitis included anterior uveitis 60%, panuveitis 24%, posterior uveitis 11%, and intermediate uveitis 6%. CONCLUSIONS: The authors found a high prevalence of acute anterior nongranulomatous uveitis. The most commonly encountered infectious uveitis included herpes virus, tuberculosis, and toxoplasmosis. PMID- 19294568 TI - Infectious uveitis in Thailand: serologic analyses and clinical features. AB - PURPOSE: To determine the seroprevalence of various infectious agents in Thai patients with uveitis. METHODS: Prospective study of 101 consecutive patients with uveitis, 100 HIV-infected retinitis patients, and 100 nonuveitis controls. RESULTS: Antibodies against T. gondii were detected in 31/101 non-HIV patients, mostly with posterior uveitis and focal retinitis, and were significantly higher than in other groups examined. Antibodies for T. pallidum and Leptospira were observed more frequently in patients with HIV-infected retinitis. Active tuberculosis in non-HIV patients was not found. CONCLUSIONS: Seroprevalence of T. gondii antibodies in patients with non-HIV posterior uveitis was higher than in nonuveitis controls and HIV patients with retinitis. PMID- 19294569 TI - Bilateral episcleritis as a manifestation of cutaneous leukocytoclastic vasculitis. AB - PURPOSE: The authors describe how bilateral episcleritis can be a sign of active systemic disease and can respond to treatment in a patient with cutaneous leukocytoclastic vasculitis. DESIGN: Case report. METHODS: Comprehensive ophthalmic and physical examination and color photography were used to monitor inflammation and its response to systemic immunosuppression. RESULTS: Systemic cyclophosphamide caused regression of systemic symptoms, cutaneous lesions, and episcleritis. After an 8-month follow-up, the patient has not had a systemic or ocular recurrence. CONCLUSION: Episcleritis may be a manifestation of cutaneous leukocytoclastic vasculitis. Careful examination of ocular inflammation is important in monitoring systemic disease and treatment. PMID- 19294570 TI - Posterior segment involvement in Korean patients with HLA-B27-associated uveitis. AB - PURPOSE: To compare clinical characteristics and visual prognosis between HLA-B27 associated uveitis patients with and without posterior segment involvement. METHODS: Medical records of 78 patients with HLA-B27-associated uveitis were retrospectively analyzed. RESULTS: Neither demographic nor clinical characteristics were associated with the presence of posterior segment involvement. In patients with posterior segment involvement, more aggressive therapeutic strategies, including immunomodulatory therapy, were frequently required. The rate of visual acuity 20/50 or worse was higher in patients with posterior involvement than in patients without posterior involvement. CONCLUSION: Patients with posterior segment involvement showed worse visual outcome than patients without posterior involvement. PMID- 19294571 TI - Herpes zoster keratouveitis and inflammatory ocular hypertension 8 years after varicella vaccination. AB - More than 8 years after varicella vaccination, a healthy 16 year-old boy presented with keratouveitis, severe inflammatory glaucoma in his left eye, and Hutchinson's sign. He was treated with systemic acyclovir, topical steroids, cycloplegics, and glaucoma medications for a full recovery two months after presentation. It is unclear whether the source of herpes zoster which reactivated in this patient represents wild type virus or his previous vaccine strain. Herpes zoster ophthalmicus is very rare in the pediatric population after varicella vaccination but can cause severe inflammatory glaucoma that requires aggressive therapy. PMID- 19294572 TI - Retinal vasculitis associated with asymptomatic Gardnerella vaginalis infection: a new clinical entity. AB - PURPOSE: To report on 3 cases of retinal vasculitis associated with asymptomatic Gardnerella vaginalis (GV) infection. METHODS: Review of 3 consecutive patients who presented with central retinal vasculitis without signs or symptoms of systemic disease. The vasculitis involved the central branches of the retinal vessels bilaterally and was steroid-dependent. RESULTS: During treatment, patients developed clinically significant vaginitis, which the gynecologist considered to be exacerbated by the steroid treatment, leading to its withdrawal. All 3 vaginal specimens were positive for GV. Antibiotic susceptibility testing led to administration of oral ampicillin (2 g/day for 10 days), which resolved both the vaginal infection and the retinal vasculitis. CONCLUSIONS: Idiopathic retinal vasculitis can be triggered by several agents. In this case series, GV was associated with retinal vasculitis, which was resolved by oral ampicillin. GV infection may be one of a number of triggers of retinal vasculitis. Appropriate treatment and full resolution of ocular inflammation requires exclusion of possible underlying infections. PMID- 19294573 TI - Fluorescein angiography and visual acuity in active uveitis with Behcet disease. AB - PURPOSE: To determine the association of fluorescein angiography characteristics with visual acuity and visual loss during medical treatment in patients with Behcet uveitis. METHODS: Ninety-three eyes from 69 Behcet patients with active panuveitis underwent fluorescein angiography, and the characteristic findings by angiography were determined. The patients were followed with immunomodulatory treatment. RESULTS: Diffuse vascular leakage (73.4%), diffuse macular leakage (66.0%), and disc leakage (52.7%) were the predominant angiographic findings on fluorescein angiography. Multivariate logistic regression analysis revealed that a macular window defect and disc neovascularization on fluorescein angiography were associated with poor visual acuity (A] p.[H3035QfsX4:V1138M] was detected, whereas her parents and another sibling were heterozygous carriers. Patient 2 had a novel homozygous nonsense mutation, c.2907C>A (p.Cys969X), in exon 21. The genotype phenotype correlation of Chinese children with novel merosin-deficient congenital muscular dystrophy is reported. PMID- 19294600 TI - Biotin-responsive basal ganglia disease: case report and review of the literature. AB - Biotin-responsive basal ganglia disease is a rare entity of which 10 cases have been reported in the literature. We report a case of biotin-responsive basal ganglia disease with similarities and differences compared to the previously reported cases by Ozand et al. Our case presented much earlier, was milder and responded better to lower doses of biotin, compared to the cases reported previously. Since our case showed differences with those in the literature, it might represent a new entity or a milder form of the same entity. PMID- 19294601 TI - Giant congenital melanocytic nevus coexistent with Chiari II malformation. AB - BACKGROUND: Giant congenital melanocytic nevus (GCMN) and Chiari II malformation (CM II) are distinct rare clinical entities. No previous report of this condition is to be found in the medical literature. MATERIAL AND METHODS: We report the case of a male infant seen initially for a GCMN and an upper thoracic myelomeningocele. MRI incidentally revealed cerebellum and brain stem elongate via the foramen magnum. The infant's physical examination included spastic quadriparesis and hypotonia. An operation for myelomeningocele repair and suboccipital craniectomy with resection of C (1) and cerebellar tonsils was done. The signs and symptoms of the patient were improved after surgery. The GCMN was not treated according to the parents' wished, but a schedule of long-term monitoring was recommended. CONCLUSION: This case report illustrates that CMNs can coexist with CM II. The clinician who diagnoses a patient with CMNs should remember that this entity can be associated with other pathologies of the central nervous system. Earlier diagnosis and surgical intervention improves the prognosis. Long-term monitoring and follow-up should be recommended for patients with CMNs. PMID- 19294602 TI - Pyridoxine-dependent epilepsy: normal outcome in a patient with late diagnosis after prolonged status epilepticus causing cortical blindness. AB - We report on a male proband with pyridoxine-dependent epilepsy (PDE) and neonatal seizure onset. At the age of 31 months, a prolonged status epilepticus led to severe neurological regression with cortical blindness, loss of speech and muscular hypotonia with slow recovery over the following 3 months. At 33 months of age pyridoxine therapy was initiated with excellent response and the boy remained seizure-free on pyridoxine monotherapy, except for two occasions with seizure recurrence 10 days after accidental pyridoxine withdrawal. alpha aminoadipic semialdehyde dehydrogenase (antiquitin) deficiency was indicated by elevated pipecolic acid concentrations in plasma and alpha-aminoadipic semialdehyde excretion in urine. Molecular analysis of the antiquitin gene revealed a novel missense mutation c.57insA, while the mutation of the other allele remained unidentified so far. Despite the delay in diagnosis and prolonged status epilepticus, neuropsychological evaluations at the ages of 11 and 18 years demonstrated full-scale IQ of 93 and 92, respectively, with better verbal IQ (103 and 101) than performance IQ (85 and 82). PMID- 19294603 TI - [Medication after hospitalization for acute coronary syndrome: Determinants of individual drug changes]. AB - BACKGROUND AND OBJECTIVE: This study examines drug changes from therapeutic medication given during hospitalization to that in further out-of-hospital treatment prescribed by medical practitioners of patients with acute coronary syndrome. The main focus of this trial was to demonstrate any change in such medication after hospital discharge. PATIENTS AND METHODS: During a six-month period a "health diary" covering health status and medication was filled in weekly by 104 patients who had been hospitalized for acute coronary syndrome in the Cologne area of Germany. The mean age of the patients was 62 years and 76 (73%) of them were men. Changes in medication between hospital discharge and further treatment were recorded. Prescribed daily doses and guideline-recommended daily doses were compared for each drug class. RESULTS: Changes in medication were not observed in a population-based approach but occurred on an individual patient's level in 40% of them when only those drugs recommended in therapeutic guidelines were analysed. A fifth more lipid-lowering drugs and over a third more beta-blockers had been prescribed than recommended in the appropriate guidelines. For other groups of drugs no significant deviations from guideline recommendations were noted. DISCUSSION: No obvious interface problem between hospital and further-treatment medication was observed in a population-based approach. However, variations in medication were found when drug prescriptions of individual patients were compared But despite these variations in individual patient the overall prescribing practice by physicians out of hospital showed good implementation of the therapeutic guidelines. PMID- 19294604 TI - [Elevated serum creatinine without discernible kidney disease]. AB - HISTORY AND PHYSICAL EXAMINATION: A 46-year-old previously healthy man was incidentally found to have an elevated serum creatinine concentration of 2,7 mg/dl (250 micromol/l) in a dry chemical enzyme test. He had no symptoms. Physical examination was unremarkable. INVESTIGATIONS: 12 days after the first laboratory test serum creatinine was to be normal (0,66 g/dl, 58 micromol/l). Urinalysis and ultrasound scan of the kidneys revealed no anomalies. But increased sarcosine levels were found in both urine and plasma. DIAGNOSIS AND COURSE: The creatinine testing assay interfered with an elevated serum sarcosine level, which is found in an inborn error of aminoacid metabolism called sarcosinemia. CONCLUSION: In a patient with sarcosinemia a dry chemical enzyme analysis which is often used by general practitioners can produce falsely high creatinine levels. When suspecting sarcosinemia, serum creatinine should be checked against another type of laboratory test. PMID- 19294605 TI - [Endogenous cardioactive steroids--a new class of steroid hormones]. PMID- 19294606 TI - [Suspected deep vein thrombosis in a 60-year-old man]. PMID- 19294607 TI - [Cardiac contractility modulation: a new option for treating systolic heart failure]. AB - Cardiac contractility modulation (CCM) is a recently developed electrical treatment for enhancing ventricular contractility by non-excitatory impulses applied during the absolute refractory period in advanced systolic heart failure, regardless of the presence of dyssynchrony and QRS prolongation. Implantation of a CCM device is comparable to pacemaker implantation. Three electrodes are placed in the heart via the subclavian vein. One electrode in the right atrium is used for sensing, while ventricular stimulation is done by two electrodes in the right ventricle. The FIX-CHF-4 trial was a randomized, double-blind and cross-over study in 164 patients with systolic heart failure and an ejection fraction 35%, in order to evaluate the effect of CCM treatment on exercise capacity and quality of life. After three months the patients' six-minute walk distance increased, while ejection fraction remained unchanged. Moreover, quality of life improved. CCM stimulation was found to be a safe method. In patients with heart failure and left ventricular systolic dysfunction exercise tolerance and quality of life may be improved by CCM. Further studies to determine morbidity and mortality during long-term CCM stimulation are indicated. PMID- 19294608 TI - [Weight reduction in obese subjects--what role do genes play?]. AB - Because of the increasing prevalence of obesity there are ever more weight-loss programmes. Independently of such programmes it is well known that there are considerable variations in weight loss under the same standardized conditions. Recent studies provide first evidence that a genetic background could be partly responsible for these variations. Significant associations were shown between specific gene variants and weight loss. However data are inconsistent and many studies have methodological limitations. Although the genetic background may influence weight loss, items such as compliance and life-style may play a greater role. Thus the current knowledge about genetic determinants of weight loss has as yet no clinical relevance. However, the identification of gene effects could contribute to offering personalized treatment of obesity and thus achieving greater weight loss. PMID- 19294609 TI - [Does the provision of patient records to a medical agency violate the confidentiality requirement? Decision of the Administrative Court in Frankfurt am Main from February 13, 2008]. PMID- 19294610 TI - [Biologic half-life of bisphosphonates]. PMID- 19294611 TI - [Which factors determine how family members experience the obsessive-compulsive disorder of a close relative?]. AB - Obsessive-compulsive disorders represent a challenge with their effects on the everyday life for family members. The main emphasis of previous research was on the recording of the burden and deficits of this group. The qualitative study on hand aims to describe living with an OCD-patient from the viewpoint of the relatives. It could be shown that the perception of the disorder is formed by the experiences of life. On an experience continuum two poles could be pointed out: on the one hand an orientation in the direction of control and on the other hand the acceptance of unchangeability. In turn these basic orientations form the view on the illness and are connected to special burdens and risks but also resources. The perception of the illness can change over time or due to situative reasons. Subjective relieving changes of the perspectives and possible influences and supports of professional helpers are of special interest for therapists. PMID- 19294612 TI - Nestin as a marker in the classification of adrenocortical tumors. AB - Expression of the intermediate filament, nestin, was long believed to be restricted to neuroectodermal stem cells. However, nestin expression has recently been detected in several tumors. Since adrenocortical carcinoma, a tumor entity still very difficult to classify, may gain the ability to aberrantly express neuroectodermal proteins including chromogranin A and synaptophysin, we asked the question whether nestin might also be detected in adrenocortical carcinomas, and if so, whether it might serve as a tool for clinical pathology. Therefore, we studied the expression of nestin in normal adrenal glands, adrenocortical adenomas, and adrenocortical cancers using specific immunohistochemistry and semi quantitative reverse transcriptase-polymerase chain reaction. Immunostaining was nestin-positive in 1 out of 9 normal adrenal glands (11%), 2 out of 20 adrenocortical adenomas (10%), and 13 out of 16 adrenocortical carcinomas (81%). Expression of nestin mRNA could be detected in all microdissected tissues, independently of their grade of dedifferentiation. We conclude that our findings provide further evidence that nestin, as a marker, is not restricted to neuronal stem cells and nestin expression is worth to be studied in adrenocortical tumors. PMID- 19294613 TI - [Survey of sensory perception and patients' subjective assessment of the application of nasal sprays - the nasal-spray-sensoric-scale]. AB - AIM: The Nasal Spray Evaluation Questionnaire is a standardized means to evaluate hedonicity of nasal sprays in the English speaking countries. To date, there is no such questionnaire for German speaking patients. The here presented study aimed at evaluating the translated questionnaire. Three nasal sprays were tested and the questionnaire was used to estimate sensory hedonicity. Finally, the questionnaire was improved and can now serve as a standardized means to measure nasal sensory perceptions. MATERIAL AND METHODS: The questionnaire was literally translated from English to German. There were three groups of 60 patients each presenting with the sensation of a 'dry nose' to a secondary medical center with a board certified otolaryngologist. The ingredients of the three different nasal sprays varied only in one phytotherapeutic addition. The first evaluation of the nasal spray was conducted immediately after application of the nasal spray, and two minutes after application. A third and optional telephone survey was performed several days later after continuous use of the spray. RESULTS: The different evaluations of nasal sprays were already lined out after the first questionnaire. It could serve as a new, reliable and reproduceable tool for the evaluation of nasal sensory perceptions. DISCUSSION: The questionnaire served as an adequate tool to estimate hedonicity of nasal sprays. First results of the German version show, however, that it could facilitate the evaluation by restructuring the questionnaire. Continuously marking good grades with high points (maximum 100) and negative answers with low points (minimum 0 points) will unify and simplify the questionnaire. CONCLUSION: The translation of the Nasal Spray Evaluation Questionnaire into a German version can serve as a new standard tool for measuring nasal sensation. PMID- 19294614 TI - [Selected performances in speech perception in children with APD]. AB - BACKGROUND: The ability to detect phonemes in spoken language (phonological processing) is equally important for first and secondary language acquisition (reading, spelling, orthography). Nevertheless, it is a subject of some controversy whether psychometric tests of auditory verbal stimuli (linguistic load) are to take in account in the diagnostics of (central) auditory processing disorders (C)APD. METHODS: Data in phonological synthesis and in verbal auditory closure, obtained from a research database of children who were audiologically and psychologically diagnosed as with auditory processing deficits, were analyzed retrospectively. These data were collected by a clinical psychologist in the diagnostic setting, who administered the subtests Sound Blending and Auditory Closure out of the German version of the Illinois Test of Psycholinguistic Abilities. PARTICIPANTS: Three clinical groups: n=51 with monosymptomatic APD; n=33 with APD+developmental language impairment; n=15 with APD and developmental dyslexia and one control group (typically developing children without clinical developmental disorders). INCLUSION CRITERIA: normal hearing status and nonverbal intelligence, monolingual German-speaking, no suspicion of attention deficit hyperactivity disorders and visual perception disorders and scoring > or = 2 SDs below the reference mean in at least 2 auditory symptoms. RESULTS: The controls showed the best performance in "Sound Blending" (T-score 49.2; SD 8.8), followed by children with monosymptomatic APD (T-score 48.0; SD 9.5) and children with co morbid conditions (APD+developmental dyslexia: T-score 45.9; SD 6.0; APD+language impairment: T-score 44.4; SD 8.7). The differences between the groups did not reach statistical significance. Test scores in "Auditory Closure" were consistently poorer in the APD-groups (children with monosymptomatic APD: T-score 50.9; SD 8.8; children with APD+developmental dyslexia: T-score 49.6; SD 7.7; children with APD+developmental language impairment: T-score 47.1; SD 10.5) than for the normal group (T-score 54.9; SD 7.5). None of the groups performed any differently from the controls. CONCLUSIONS: Because the acoustic-verbal automatic dimensions of sound blending and auditory closure did not differentiate the 4 study groups, experts should renounce of them in the diagnostics of (C)APDs. PMID- 19294615 TI - Laparoscopic cholecystectomy without visible scar. Combined suprapubic and transumbilical approach: the "Minden cholecystectomy". PMID- 19294616 TI - Ewing's sarcoma / primitive neuroectodermal tumor of the kidney. AB - A 42-year-old female presented with right back pain. The CT scan revealed a 72-mm space-occupying lesion in the middle portion of the right kidney. No metastasis was proven. She underwent laparoscopic radical nephrectomy and lymph node disection. The histopathological examination revealed a high-grade primitive small round tumor the cells of which were strongly positive for CD99 and vimentin. Fluorescence in situ hybridization analysis using a DNA probe for the Ewing sarcoma breakpoint region 1 (EWSR 1) on chromosome 22g12 revealed a rearrangement of the EWSR 1 locus. The diagnosis was Ewing's sarcoma / primitive neuroectodermal tumor of the kidney. She underwent 13 cycles of chemotherapy, and has no evidence of recurrence 19 months after surgery. PMID- 19294617 TI - [Dietary intake of non-dioxin-like polychlorinated biphenyls (PCB) in Bavaria, Germany. Results from the Integrated Exposure Assessment Survey (INES)]. AB - Polychlorinated biphenyls (PCB) were widely used in numerous industrial and commercial applications in high quantities in the past. Based on their persistence in the environment, their tendency to accumulate in the organism and their specific health effects, PCBs have to be assessed as critical substances. Because the dietary intake was assumed to be the main intake route, the Integrated Exposure Assessment Survey (INES) aimed to measure the recent exposure to PCBs in Germany. The study consisted of 10 female and 10 male participants living in Munich and surroundings. The participants collected dietary duplicates of all food consumed and prepared as for consumption over 7 consecutive days. Altogether the 6 non-dioxin-like PCB congeners 28, 52, 101, 138, 153 and 180 ndl PCB or, respectively, indicator PCB and furthermore the congener 118 were detected using a gas chromatographic method. Dietary intake was calculated using the amount of food eaten daily and the results from the duplicates. Using the sum of PCB 138, 153 and 180 multiplied by 4, the daily intake ranged from 4.0 to 24.1 ng/kg b.w. (median: 9.5 ng/kg b.w.). On the contrary, the daily intake was 2.9 to 20.6 ng/kg b.w. (median: 11.2 ng/kg b.w.) if the sum of the 6 indicator PCBs multiplied by factor 2 was used for quantification. No sex-related difference of the dietary intake was observable. Overall, it can be concluded that the dietary PCB intake has further decreased in the last years in Germany. At present, the toxicological database is not suitable to assess the risks coming solely from the non-dioxin-like PCBs because it is not possible to differentiate between non dioxin-like and dioxin-like effects in toxicological studies. Nevertheless, a further reduction of PCB exposure via food by searching for possible sources is needed. PMID- 19294618 TI - [Case report--surgical therapy of a retroperitoneal liposarcoma weighing 45 kg]. AB - Due to the late onset of symptoms, retroperitoneal liposarcoma are often diagnosed in advanced stages when adjacent organs have been infiltrated and the tumours have reached extensive sizes. Surgery remains the first choice of therapy. We report on the primary resection of a 45-kg liposarcoma that was removed en-bloc including the left kidney and descending colon with -tumour-free margins. Nine months later, the follow-up revealed a right-sided recurrence of the tumour, which was surgically removed including the right ureter. Since then, the patient has been without any signs of tumour recurrence or metastases. This report demonstrates that even extreme-ly large tumours can be removed safely and that the size is not a contraindication for primary surgical treatment. Local recurrence is common as seen in our case, and occurs even after R0 resection up to 10 years after the first operation. Recurrences should be surgically removed as this is the only treatment which has been shown to increase survival in even R1 and R2 situations. PMID- 19294619 TI - [Do we need a better method for diagnosing coma?]. PMID- 19294621 TI - Proteome wide screening using peptide affinity capture. AB - MS-based strategies are key technologies for identifying proteins in proteomic research. Despite significant improvements in recent years efficient fractionation processes of target analytes remain major bottlenecks in MS-based protein analysis. Immunoaffinity-based sample fractionation strategies have shown their potential for the enrichment of analyte peptides of interest, but only small numbers of analytes can be quantified in one experiment. The lack of appropriate capture reagents limits the application of immunoaffinity-based approaches and only biased biomarker discovery approaches are possible. This perspective discusses the current status of immunoaffinity MS-based approaches and introduces a novel concept that uses group specific anti-peptide antibodies - Triple X Proteomics Antibodies -- for the enrichment of signature peptides. Classes of peptides with identical termini can be fractionated based on TXP immunoaffinity enrichment steps and can subsequently be identified using established tandem MS procedures. Based on bioinformatic algorithms minimal sets of TXP epitopes can be specified, that cover a wide range of given proteome landscapes of one or even several different species. This opens the possibility to use a minimal number of TXP antibodies as a universal toolbox for general immunoaffinity-based approaches in proteome analysis. PMID- 19294623 TI - Affinity-MS: methods and applications in proteomics research. PMID- 19294624 TI - Radiological findings of early invasive pulmonary aspergillosis in immune compromised patients. AB - Data on the radiological features of invasive pulmonary aspergillosis (IPA) in early stages is scanty. Detection of Aspergillus (ASP) species in broncho alveolar (BAL) fluid by polymerase chain reaction (PCR) enables early diagnosis of IPA. This study describes the radiological features of early stages of IPA. Chest computerized tomography (CT) films of 22 consecutive immune-compromised patients with IPA diagnosed with the aid of ASP PCR testing from BAL fluid were characterized and compared to that of 18 similar patients diagnosed with traditional bacteriological methods and to data from the literature. It was found that patients diagnosed with the aid of ASP PCR testing tended to have focal disease as manifested by more 11-30 mm nodules with halo (68% vs. 33%, p = 0.04), more focal ground glass (single area 32% vs. 6%, p = 0.05, patchy 32% vs. 0%, p = 0.01) and less diffuse ground glass (0% vs. 22%, p = 0.03), less cavitations (5% vs. 28%, p = 0.05) and less consolidations (segmental 14% vs. 50%, p = 0.02 and diffuse 14% vs. 67%, p = 0.001). It was concluded that the radiological appearance of early IPA diagnosed with the aid of PCR testing included mainly discrete small nodules with halo and focal ground glass, representing the early stage of the disease. PMID- 19294625 TI - Quantitative analysis of cell signaling and drug action via mass spectrometry based systems level phosphoproteomics. AB - Protein phosphorylation is a primary form of information transfer in cell signaling pathways and plays a crucial role in regulating biological responses. Aberrant phosphorylation has been implicated in a number of diseases, and kinases and phosphatases, the cellular enzymes that control dynamic phosphorylation events, present attractive therapeutic targets. However, the innate complexity of signaling networks has presented many challenges to therapeutic target selection and successful drug development. Approaches in phosphoproteomics can contribute functional, systems-level datasets across signaling networks that can provide insight into suitable drug targets, more broadly profile compound activities, and identify key biomarkers to assess clinical outcomes. Advances in MS-based phosphoproteomics efforts now provide the ability to quantitate phosphorylation with throughput and sensitivity to sample a significant portion of the phosphoproteome in clinically relevant systems. This review will discuss recent work and examples of application data that demonstrate the utility of MS, with a particular focus on the use of quantitative phosphoproteomics and phosphotyrosine directed signaling analyses to provide robust measurement for functional biological interpretation of drug action on signaling and phenotypic outcomes. PMID- 19294626 TI - Immunotherapy with cytokine induced killer cells in solid and hematopoietic tumours: a pilot clinical trial. AB - BACKGROUND AND OBJECTIVES: CIK cells are a novel population of efficient immune effector cells with high antitumour activity mainly due to the high proliferation of CD3(+)CD56(+) cells, so may play a role in the development of new forms of adoptive cellular immunotherapy. We started a pilot clinical trial with autologous CIK cells in patients with refractory lymphoma and metastatic solid tumours. This study was aimed at determining the feasibility of generating a sufficient number of CIK cells in heavily pretreated patients and at assessing treatment toxicity. DESIGN AND METHODS: CIK cells were generated from peripheral blood mononuclear cells (MNC) and incubated in the presence of IFN-gamma followed by OKT3 and IL-2. Treatment schedule consisted of three cycles of CIK cells infusions at an interval of 3 weeks. RESULTS: At present 12 patients were enrolled: 6 advanced lymphomas, 5 metastatic kidney carcinoma and 1 hepatocellular carcinoma (HCC). The median number of transferred cells per patient was 28 x 10(9) (range, 6-61). Protocol adherence was excellent and the toxicity profile was favourable. After CIK cells infusion, the absolute median count of lymphocytes, CD3(+), CD8(+) and CD3(+)CD56(+) cells significantly increased in patient's peripheral blood. Clinical outcome appeared promising: three patients had complete response (CR) and two patients had stabilization of disease with a median follow-up of 33 months (range, 9-44). INTERPRETATIONS AND CONCLUSIONS: These preliminary data showed that adoptive immunotherapy with CIK cells is a safe therapy with some suggestion of efficacy that significantly enhances immune functions increasing absolute numbers of effector cells without side effects. If confirmed in larger scale studies, these promising results may have a favourable impact on conventional treatment strategy of malignancies. PMID- 19294627 TI - Nitromatrix provides improved LC-MALDI signals and more protein identifications. AB - The beneficial use of NC in MALDI-MS has previously been reported to provide better S/N and reproducibility as well as less alkali metal adducts. We have therefore investigated if additional beneficial properties of NC also existed for commonly employed proteomics-based LC-MALDI procedures. Specifically we studied the effects of NC as a matrix cofactor for prestructured sample supports (AnchorChip plates), and compared the performance with several alternative sample preparation methods recently reported in the literature. The work reported here describes a new method of mixing the NC-matrix solution with the LC-eluent prior to sample deposition and shows that a mixture of CHCA and NC in a complex solvent offers superior analytical results in several ways: most striking is the higher signal intensity, and that the signals last much longer, due to the robustness of the matrix formulation. We have tested the use of the nitromatrix on a single LC MALDI preparation and found that at least ten reiterative analyses could be performed, resulting in total analysis times of more than 75 h (approximately 15 million laser shots). Consequently more than twice as many proteins could be identified than from a single analysis. This combination of longer, and stronger, MALDI signals provided an increase in the number of peptides, greater sequence coverage in MS/MS experiments and ultimately more confident peptide assignments. PMID- 19294628 TI - The detection, correlation, and comparison of peptide precursor and product ions from data independent LC-MS with data dependant LC-MS/MS. AB - The detection, correlation, and comparison of peptide and product ions from a data independent LC-MS acquisition strategy with data dependent LC-MS/MS is described. The data independent mode of acquisition differs from an LC-MS/MS data acquisition since no ion transmission window is applied with the first mass analyzer prior to collision induced disassociation. Alternating the energy applied to the collision cell, between low and elevated energy, on a scan-to-scan basis, provides accurate mass precursor and associated product ion spectra from every ion above the LOD of the mass spectrometer. The method therefore provides a near 100% duty cycle, with an inherent increase in signal intensity due to the fact that both precursor and product ion data are collected on all isotopes of every charge-state across the entire chromatographic peak width. The correlation of product to precursor ions, after deconvolution, is achieved by using reconstructed retention time apices and chromatographic peak shapes. Presented are the results from the comparison of a simple four protein mixture, in the presence and absence of an enzymatically digested protein extract from Escherichia coli. The samples were run in triplicate by both data dependant analysis (DDA) LC-MS/MS and data-independent, alternate scanning LC-MS. The detection and identification of precursor and product ions from the combined DDA search results of the four protein mixture were used for comparison to all other data. Each individual set of data-independent LC-MS data provides a more comprehensive set of detected ions than the combined peptide identifications from the DDA LC-MS/MS experiments. In the presence of the complex E. coli background, over 90% of the monoisotopic masses from the combined LC-MS/MS identifications were detected at the appropriate retention time. Moreover, the fragmentation pattern and number of associated elevated energy product ions in each replicate experiment was found to be very similar to the DDA identifications. In the case of the corresponding individual DDA LC-MS/MS experiment, 43% of the possible detectable peptides of interest were identified. The presented data illustrates that the time-aligned data from data-independent alternate scanning LC-MS experiments is highly comparable to the data obtained via DDA. The obtained information can therefore be effectively and correctly deconvolved to correlate product ions with parent precursor ions. The ability to generate precursor product ion tables from this information and subsequently identify the correct parent precursor peptide will be illustrated in a companion manuscript. PMID- 19294629 TI - Database searching and accounting of multiplexed precursor and product ion spectra from the data independent analysis of simple and complex peptide mixtures. AB - A novel database search algorithm is presented for the qualitative identification of proteins over a wide dynamic range, both in simple and complex biological samples. The algorithm has been designed for the analysis of data originating from data independent acquisitions, whereby multiple precursor ions are fragmented simultaneously. Measurements used by the algorithm include retention time, ion intensities, charge state, and accurate masses on both precursor and product ions from LC-MS data. The search algorithm uses an iterative process whereby each iteration incrementally increases the selectivity, specificity, and sensitivity of the overall strategy. Increased specificity is obtained by utilizing a subset database search approach, whereby for each subsequent stage of the search, only those peptides from securely identified proteins are queried. Tentative peptide and protein identifications are ranked and scored by their relative correlation to a number of models of known and empirically derived physicochemical attributes of proteins and peptides. In addition, the algorithm utilizes decoy database techniques for automatically determining the false positive identification rates. The search algorithm has been tested by comparing the search results from a four-protein mixture, the same four-protein mixture spiked into a complex biological background, and a variety of other "system" type protein digest mixtures. The method was validated independently by data dependent methods, while concurrently relying on replication and selectivity. Comparisons were also performed with other commercially and publicly available peptide fragmentation search algorithms. The presented results demonstrate the ability to correctly identify peptides and proteins from data independent acquisition strategies with high sensitivity and specificity. They also illustrate a more comprehensive analysis of the samples studied; providing approximately 20% more protein identifications, compared to a more conventional data directed approach using the same identification criteria, with a concurrent increase in both sequence coverage and the number of modified peptides. PMID- 19294631 TI - The influence of dioxygen on luminol chemiluminescence. AB - Assays of peroxy compounds are commonly performed after chromatographic separation of analysed mixtures. In high-performance liquid chromatography (HPLC), solvent reservoirs are sparged by helium or inline vacuum-degassed in order to control the compressibility of the solvents for efficient pumping. In this study, we investigated the influence of degassing the reaction solution on the light output of the hemin-catalyzed luminol oxidation by various oxidants. We found that, when t-butyl hydroperoxide, hydrogen peroxide, n-butyl hydroperoxide, iodosobenzene and iodobenzene diacetate were used as oxidants, the luminol chemiluminescence was lowered by 50-70% compared with an equilibrated and degassed solution. The opposite effect was observed when dibenzoyl peroxide and 3 chloroperoxybenzoic acid were used as oxidants, as the chemiluminescence increased by approximately 20-30%. The reduced chemiluminescence was explained based on the known role of dioxygen in luminol chemiluminescence. The enhancement of chemiluminescence was rationalized by suggesting an alternative mechanism of luminol oxidation valid for peroxyacids and diacyl peroxides in which the reaction of a peroxyacid anion with the diazaquinone led to light emission with a higher quantum yield than the usual path, which is suppressed by the removal of dioxygen from the reaction solution. PMID- 19294632 TI - First-trimester Down syndrome screening performance in the Dutch population; how to achieve further improvement? AB - OBJECTIVE: To study the performance of the first-trimester combined test between 2004 and 2006 compared to a previous period to investigate changes in time and identify reasons for sub-optimal performance. METHODS: Serum samples were analysed for pregnancy-associated plasma protein A (PAPP-A) and the free beta subunit of human chorionic gonadotrophin (f beta-hCG). Nuchal translucency (NT) was measured between 10 and 14 weeks. Tests were considered screen positive, if their calculated Down syndrome (DS) risk was at least 1 in 250 at term. RESULTS: A total of 20,293 singleton pregnancies were included in the analysis. The median maternal age fell from 35.7 to 34.3 years. The overall median weight-corrected multiple of the median (MoM) values of PAPP-A and f beta-hCG were 1.12 and 1.03, respectively. The median MoM value of NT was 0.89 and increased from 0.82 to 0.96. Sixty-six DS cases were detected by the screening test. The detection rate (DR) for DS was 75.9%, with a FPR of 3.3%. CONCLUSION: The performance of the first-trimester test has improved over the years. A better performance of the NT measurement was the main reason, although NT assessment should further be improved. In addition, a better setting of the medians for the biochemical parameters may contribute to a higher DR. PMID- 19294633 TI - On what diseases and health conditions should new economic research on health and development focus? AB - Given the public goods nature of research, economic research on health in developing countries is likely to have the highest returns by focusing, inter alia, on diseases and health conditions that are relatively widespread and costly and that are relatively rapidly growing. This article first summarizes the time patterns in economic research on diseases and health in developing countries for 1990-2005. It then compares those time patterns with the distribution of disability-adjusted life years (DALYs) for diseases and health conditions in developing countries estimated for 2005 and for 2030. These comparisons suggest relatively overemphasis on HIV/AIDS and underemphasis on noncommunicable diseases (NCDs). This opens the possibility for individuals or organizations initiating, re-evaluating, or increasing their economic research on health and development to make a significant contribution by focusing particularly on the analysis of behaviour and policy choices related to NCDs. Careful consideration must, of course, be given to other demands, but on the basis of these two criteria, potential contributions are likely to be greatest from research with such a focus. PMID- 19294634 TI - The growth of poor children in China 1991-2000: why food subsidies may matter. AB - How did rapid growth in per capita income and rising income inequality during 1991-2000 in China affect the health status of Chinese children, given that the disappearance in the 1990s of subsidized food coupons simultaneously increased the importance of money income in enabling consumption of basic foods by poor families? Using the China Health and Nutrition Survey data for 1991, 1993, 1997, and 2000 on 4400 households in nine provinces, we examine the height-for-age of Chinese children aged 2-13, with particular emphasis on the growth of children living in poor households. We use mean regression and quantile regression models to isolate the dynamic impact of poverty status and food coupon use on child height-for-age. Our principal findings are: (i) controlling for standard variables (e.g. parents' weight, height, and education) poverty is correlated with slower growth in height-for-age between 1997 and 2000 but not earlier; (ii) in 2000, poverty is negatively correlated with strong growth in height-for-age; and (iii) food coupon use in earlier periods correlates positively with growth in height-for-age. The general moral is the crucial social protection role that subsidized food programmes can potentially play in maintaining the health of poor children. PMID- 19294635 TI - Malnutrition, poverty, and economic growth. AB - This paper argues that indicators of anthropometric shortfall - especially low height and low weight-for-age - are uniquely suited for assessing absolute deprivation in developing countries. Anthropometric indicators are relatively precise, readily available for most countries, reflect the preferences and concerns of many poor people, consistent with reckoning the phenomenon directly in the space of functionings, intuitive, easy to use for advocacy, and consistent over time and across subgroups. Anthropometric indicators can therefore complement (but not replace) standard indicators of income/consumption poverty, especially for comparisons across subgroups, within households, across countries, and in the long run. In addition, the paper analyses spells of change in malnutrition over time, finding that the association between economic growth and chronic child malnutrition is very small (but statistically significant) and much lower than the elasticity of growth on poverty. The policy implication of this finding is that direct interventions aimed at reducing infant malnutrition are required. PMID- 19294636 TI - Measuring health inequality with realization of potential life years (RePLY). AB - This paper proposes a new method to measure health inequalities that are caused by conditions amenable to policy intervention. The method is built on a technique that can separate avoidable and unavoidable mortality risks, using world mortality data compiled by the World Health Organization for the year 2000. The new method is applied to data from 191 countries. It is found that controlling for unavoidable mortality risks leads to a lower estimate of health inequality than otherwise, especially for developed countries. Furthermore, although countries with a higher life expectancy at birth tend to have lower health inequality, there are significant variations in health inequalities across countries with the same life expectancy. The results therefore support the WHO's plea for using health inequality as a distinct parameter from the average level of health in assessing the performance of health systems. PMID- 19294637 TI - Life expectancy and welfare in Latin America and the Caribbean. AB - This paper analyses the recent evolution of life expectancy in Latin American and Caribbean countries, and evaluates how much it has contributed to the overall improvements in welfare. We argue that increases in life expectancy between 1960 and 2000, which were largely independent of income, represented gains in welfare comparable to the ones derived from income growth. For countries in the region, estimates of welfare improvements accounting for health increase the numbers obtained from income alone by 40% on average. The available evidence suggests that improvements in public health infrastructure - such as provision of treated water and sewerage services - and large-scale immunization programs may have been the key factors behind the mortality reductions observed in the period. PMID- 19294638 TI - Measuring intra-household health inequality: explorations using the body mass index. AB - This paper examines the relationship between level of well-being and inequality at inter-country and intra-household levels, using individuals' body mass index (BMI) rather than income as the indicator of well-being. BMI is useful for these purposes because (1) it is measured at the individual rather than household level; (2) it reflects command over food, but also non-food resources that affect health status like sanitary conditions and labour-saving technologies; (3) it accounts for caloric consumption relative to needs; (4) it is easily measured; and (5) any measurement error is likely to be random. We do not find any evidence to support the idea of an intra-household or inter-country Kuznets curve. We study the correlations between average household well-being, still measured by BMI, and differences in the BMIs of males and females, parents and children. Here, we find a tendency to protect the BMI of young children when living standards are very low. We find no clear patterns by gender. Perhaps the most striking finding in the paper is that about half of total BMI inequality at the country level is within households. Thus, standard measures of inequality that use household-level data may drastically understate true inequality. PMID- 19294639 TI - Health inequality and deprivation. AB - This paper looks at health inequality and deprivation, with a particular focus on developing countries. It is specifically concerned with relationships between health and income, especially the extent to which inequality and deprivation in the former is driven by changes in the latter. The paper reports increasing disparity in child mortality among country groups since the mid-1970s. It also reports decreased inequality in life expectancy among countries from the early 1960s until the late 1980s and increased inequality thereafter. Similar patterns in life expectancy deprivation are reported. The paper finds that this is partly due to a changing behavioural relationship between life expectancy and income per capita among countries with low achievement in the former variable. The paper also introduces and provides an overview of the papers that follow in this Supplement. PMID- 19294640 TI - Heritability of regional and global brain structure at the onset of puberty: a magnetic resonance imaging study in 9-year-old twin pairs. AB - Puberty represents the phase of sexual maturity, signaling the change from childhood into adulthood. During childhood and adolescence, prominent changes take place in the brain. Recently, variation in frontal, temporal, and parietal areas was found to be under varying genetic control between 5 and 19 years of age. However, at the onset of puberty, the extent to which variation in brain structures is influenced by genetic factors (heritability) is not known. Moreover, whether a direct link between human pubertal development and brain structure exists has not been studied. Here, we studied the heritability of brain structures at 9 years of age in 107 monozygotic and dizygotic twin pairs (N = 210 individuals) using volumetric MRI and voxel-based morphometry. Children showing the first signs of secondary sexual characteristics (N = 47 individuals) were compared with children without these signs, based on Tanner-stages. High heritabilities of intracranial, total brain, cerebellum, and gray and white matter volumes (up to 91%) were found. Regionally, the posterior fronto occipital, corpus callosum, and superior longitudinal fascicles (up to 93%), and the amygdala, superior frontal and middle temporal cortices (up to 83%) were significantly heritable. The onset of secondary sexual characteristics of puberty was associated with decreased frontal and parietal gray matter densities. Thus, in 9-year-old children, global brain volumes, white matter density in fronto occipital and superior longitudinal fascicles, and gray matter density of (pre )frontal and temporal areas are highly heritable. Pubertal development may be directly involved in the decreases in gray matter areas that accompany the transition of our brains from childhood into adulthood. PMID- 19294641 TI - Cortical mapping of naming errors in aphasia. AB - Persons with aphasia vary greatly with regard to clinical profile; yet, they all share one common feature-anomia-an impairment in naming common objects. Previous research has demonstrated that particular naming errors are associated with specific left hemisphere lesions. However, we know very little about the cortical activity in the preserved brain areas that is associated with aphasic speech errors. Utilizing functional magnetic resonance imaging (fMRI), we show for the first time that specific speech errors are associated with common cortical activity in different types and severities of aphasia. Specifically, productions of phonemic errors recruited the left posterior perilesional occipital and temporal lobe areas. A similar pattern of activity was associated with semantic errors, albeit in the right hemisphere. This study does not discount variability in cortical activity following left hemisphere stroke; rather, it highlights commonalities in brain modulation in a population of patients with a common diagnosis but vastly different clinical profiles. PMID- 19294642 TI - Detection of irregular, transient fMRI activity in normal controls using 2dTCA: comparison to event-related analysis using known timing. AB - When events occur spontaneously during the acquisition of a series of images, traditional modeling methods for detecting functional MRI activation detection cannot be employed. The two-dimensional temporal clustering algorithm, 2dTCA, has been shown to accurately detect random, transient activations in computer simulations without the use of known event timings. In this study we applied the 2dTCA technique to detect the timings and spatial locations of sparse, irregular, transient activations of the visual, auditory, and motor cortices in 12 normal controls. Experiments with one and two independent types of stimuli were employed. Event-related activation using known timing was compared with event related activation using 2dTCA-detected timing in individuals and across groups. The 2dTCA algorithm detected the activation from all presented stimuli in every subject. When compared with block-design results using a measure of correlation between activation maps, no significant difference was found between the 2dTCA activation maps and the event-related maps using known timing across all subjects. Therefore, 2dTCA has the potential to be an accurate and more practical method for detection of spontaneous, transient events using fMRI. PMID- 19294643 TI - Death and survival of heterozygous Lurcher Purkinje cells in vitro. AB - The differentiation and survival of heterozygous Lurcher (+/Lc) Purkinje cells in vitro was examined as a model system for studying how chronic ionic stress affects neuronal differentiation and survival. The Lurcher mutation in the delta2 glutamate receptor (GluRdelta2) converts an orphan receptor into a membrane channel that constitutively passes an inward cation current. In the GluRdelta2(+/Lc) mutant, Purkinje cell dendritic differentiation is disrupted and the cells degenerate following the first week of postnatal development. To determine if the GluRdelta2(+/Lc) Purkinje cell phenotype is recapitulated in vitro, +/+, and +/Lc Purkinje cells from postnatal Day 0 pups were grown in either isolated cell or cerebellar slice cultures. GluRdelta2(+/+) and GluRdelta2(+/Lc) Purkinje cells appeared to develop normally through the first 7 days in vitro (DIV), but by 11 DIV GluRdelta2(+/Lc) Purkinje cells exhibited a significantly higher cation leak current. By 14 DIV, GluRdelta2(+/Lc) Purkinje cell dendrites were stunted and the number of surviving GluRdelta2(+/Lc) Purkinje cells was reduced by 75% compared to controls. However, treatment of +/Lc cerebellar cultures with 1-naphthyl acetyl spermine increased +/Lc Purkinje cell survival to wild type levels. These results support the conclusion that the Lurcher mutation in GluRdelta2 induces cell autonomous defects in differentiation and survival. The establishment of a tissue culture system for studying cell injury and death mechanisms in a relatively simple system like GluRdelta2(+/Lc) Purkinje cells will provide a valuable model for studying how the induction of a chronic inward cation current in a single cell type affects neuronal differentiation and survival. PMID- 19294644 TI - Adult neurogenesis in the crayfish brain: proliferation, migration, and possible origin of precursor cells. AB - The birth of new neurons and their incorporation into functional circuits in the adult brain is a characteristic of many vertebrate and invertebrate organisms, including decapod crustaceans. Precursor cells maintaining life-long proliferation in the brains of crayfish (Procambarus clarkii, Cherax destructor) and clawed lobsters (Homarus americanus) reside within a specialized niche on the ventral surface of the brain; their daughters migrate to two proliferation zones along a stream formed by processes of the niche precursors. Here they divide again, finally producing interneurons in the olfactory pathway. The present studies in P. clarkii explore (1) differential proliferative activity among the niche precursor cells with growth and aging, (2) morphological characteristics of cells in the niche and migratory streams, and (3) aspects of the cell cycle in this lineage. Morphologically symmetrical divisions of neuronal precursor cells were observed in the niche near where the migratory streams emerge, as well as in the streams and proliferation zones. The nuclei of migrating cells elongate and undergo shape changes consistent with nucleokinetic movement. LIS1, a highly conserved dynein-binding protein, is expressed in cells in the migratory stream and neurogenic niche, implicating this protein in the translocation of crustacean brain neuronal precursor cells. Symmetrical divisions of the niche precursors and migration of both daughters raised the question of how the niche precursor pool is replenished. We present here preliminary evidence for an association between vascular cells and the niche precursors, which may relate to the life-long growth and maintenance of the crustacean neurogenic niche. PMID- 19294645 TI - Management of pregnancy in a carrier of the Donohue mutation. PMID- 19294646 TI - Beta-amyloid protein (25-35) disrupts hippocampal network activity: role of Fyn kinase. AB - Early cognitive deficit characteristic of early Alzheimer's disease seems to be produced by the soluble forms of beta-amyloid protein. Such cognitive deficit correlates with neuronal network dysfunction that is reflected as alterations in the electroencephalogram of both Alzheimer patients and transgenic murine models of such disease. Correspondingly, recent studies have demonstrated that chronic exposure to betaAP affects hippocampal oscillatory properties. However, it is still unclear if such neuronal network dysfunction results from a direct action of betaAP on the hippocampal circuit or it is secondary to the chronic presence of the protein in the brain. Therefore, we aimed to explore the effect of acute exposure to betaAP(25-35) on hippocampal network activity both in vitro and in vivo, as well as on intrinsic and synaptic properties of hippocampal neurons. We found that betaAP(25-35), reversibly, affects spontaneous hippocampal population activity in vitro. Such effect is not produced by the inverse sequence betaAP(35 25) and is reproduced by the full-length peptide betaAP(1-42). Correspondingly betaAP(25-35), but not the inverse sequence betaAP(35-25), reduces theta-like activity recorded from the hippocampus in vivo. The betaAP(25-35)-induced disruption in hippocampal network activity correlates with a reduction in spontaneous neuronal activity and synaptic transmission, as well as with an inhibition in the subthreshold oscillations produced by pyramidal neurons in vitro. Finally, we studied the involvement of Fyn-kinase on the betaAP(25-35) induced disruption in hippocampal network activity in vitro. Interestingly, we found that such phenomenon is not observed in slices obtained from Fyn-knockout mice. In conclusion, our data suggest that betaAP acutely affects proper hippocampal function through a Fyn-dependent mechanism. We propose that such alteration might be related to the cognitive impairment observed, at least, during the early phases of Alzheimer's disease. PMID- 19294647 TI - Structural plasticity of dentate granule cell mossy fibers during the development of limbic epilepsy. AB - Altered granule cell>>CA3 pyramidal cell synaptic connectivity may contribute to the development of limbic epilepsy. To explore this possibility, granule cell giant mossy fiber bouton plasticity was examined in the kindling and pilocarpine models of epilepsy using green fluorescent protein-expressing transgenic mice. These studies revealed significant increases in the frequency of giant boutons with satellite boutons 2 days and 1 month after pilocarpine status epilepticus, and increases in giant bouton area at 1 month. Similar increases in giant bouton area were observed shortly after kindling. Finally, both models exhibited plasticity of mossy fiber giant bouton filopodia, which contact GABAergic interneurons mediating feedforward inhibition of CA3 pyramids. In the kindling model, however, all changes were fleeting, having resolved by 1 month after the last evoked seizure. Together, these findings demonstrate striking structural plasticity of granule cell mossy fiber synaptic terminal structure in two distinct models of adult limbic epileptogenesis. We suggest that these plasticities modify local connectivities between individual mossy fiber terminals and their targets, inhibitory interneurons, and CA3 pyramidal cells potentially altering the balance of excitation and inhibition during the development of epilepsy. PMID- 19294648 TI - Patterns of retrograde amnesia for recent and remote incidental spatial learning in rats. AB - A non-navigational test of incidental spatial learning was used to determine whether hippocampal damage causes temporally-graded retrograde amnesia (TGRA) for allocentric-spatial information. Rats were exposed to two identical objects in a circular open field for 7 min on seven consecutive days. In the 1-3 days after the last day of familiarization, rats received neurotoxic lesions of the hippocampal formation (HPC) or sham lesions. Another two groups received the same lesions 3 weeks after familiarization. The rats were then placed back in the open field with one object displaced, and the time spent in each of the quadrants as well as time spent exploring the objects was recorded. Rats that received HPC lesions 3 weeks but not 1-3 days after familiarization showed evidence of preserved remote spatial memory; however, their remote memory was expressed through different behavior than control rats. Rats with HPC lesions spent more time with the displaced object than with the object that remained in the same place, whereas control rats spent more time in the quadrant where the displaced object used to be. These results suggest that remote spatial memories may be preserved with a sufficiently long familiarization-to-surgery interval before HPC lesions, but that the nature of these memories may differ in quantity and/or quality from those of intact rats. PMID- 19294649 TI - Hippocampal long-term potentiation is enhanced in urethane-anesthetized RGS2 knockout mice. AB - RGS2 is a member of the regulator of G-protein signaling (RGS) family and has been implicated in cellular mechanisms associated with neuronal plasticity. Long term potentiation (LTP) of RGS2 knockout and wild-type mice was examined at the Schaffer collaterals to CA1 pathway in urethane-anesthetized mice in vivo to examine RGS2's possible role in the regulation of potentiation. As compared to wild-type mice, RGS2 knockouts demonstrated much stronger LTP of the extracellular population spikes at the somatic and dendritic layers in CA1 region and more pronounced LTP of the population excitatory postsynaptic current sink. Under baseline conditions, RGS2 knockouts showed lower paired-pulse facilitation of the excitatory postsynaptic potentials and associated current sinks in vivo as compared with wild-type mice. The data show for the first time that RGS2 deficient mice in vivo differ from wild-type mice in both short-term and long term synaptic plasticity suggesting that RGS2 serves as a negative regulator of long-term synaptic plasticity. PMID- 19294650 TI - Running exercise-induced up-regulation of hippocampal brain-derived neurotrophic factor is CREB-dependent. AB - The past decade has witnessed burgeoning evidence that antidepressant medications and physical exercise increase the expression of hippocampal brain-derived neurotrophic factor (BDNF). This phenomenon has gained widespread appeal, because BDNF is one of the first macromolecules observed to play a central role not only in the treatment of mood disorders, but also in neuronal survival-, growth-, and plasticity-related signaling cascades. Thus, it has become critical to understand how BDNF synthesis is regulated. Much evidence exists that changes in BDNF expression result from the activation/phosphorylation of the transcription factor, cAMP-response-element binding protein (CREB) following the administration of antidepressant medications. Utilizing a mouse model genetically engineered with an inducible CREB repressor, our current study provides evidence that increases in BDNF expression and cellular survival signaling resulting from physical exercise are also dependent upon activation of this central transcription factor. The transcription and expression of hippocampal BDNF, as well as the activation of Akt, a key survival signaling molecule, were measured following acute exercise, and also following short-term treatment with the norepinephrine reuptake inhibitor, reboxetine. We found that both interventions led to a marked increase in hippocampal BDNF mRNA, BDNF protein, and Akt phosphorylation (as well as CREB phosphorylation) in wild-type mice. As expected, activation of the CREB repressor in mutant mice sharply decreased CREB phosphorylation. In addition, all measures noted above remained at baseline levels when mutant mice exercised or received reboxetine. Increases in BDNF and phospho-Akt were also prevented when mutant mice received a combination of exercise and antidepressant treatment. The results are discussed in the context of what is currently known about BDNF signaling. PMID- 19294651 TI - Neural connectivity as an intermediate phenotype: brain networks under genetic control. AB - Recent evidence suggests that default mode connectivity characterizes neural states that account for a sizable proportion of brain activity and energy expenditure, and therefore represent a plausible neural intermediate phenotype. This implies the possibility of genetic control over systems-level connectivity features. Imaging genetics is an approach to combine genetic assessment with multimodal neuroimaging to discover neural systems linked to genetic abnormalities or variation. In the present contribution, we report results obtained from applying this strategy to both structural connectivity and functional connectivity data. Using data for serotonergic (5-HTTLPR, MAO-A) and dopaminergic (DARPP-32) genes as examples, we show that systems-level connectivity networks under genetic control can be identified. Remarkable similarities are observed across modalities and scales of description. Features of connectivity often better account for behavioral effects of genetic variation than regional parameters of activation or structure. These data provide convergent evidence for genetic control in humans over connectivity systems, whose characterization has promise for identifying neural systems mediating genetic risk for complex human behavior and psychiatric disease. PMID- 19294652 TI - Common and specific contributions of the intraparietal sulci to numerosity and length processing. AB - Numerical and spatial magnitude processing have long been intimately associated, leading to the suggestion that they share a common system of magnitude representation. Although separate investigations on the cerebral areas involved in numerosity and spatial estimation point toward the parietal cortex, the precise anatomical overlap, if any, has not yet been directly established. Here, functional magnetic resonance imaging was used to localize the cerebral network involved in processing both numerosity and length. Blood oxygenation level dependent signal changes were measured while healthy volunteers were making numerosity comparisons on linear arrays of dots, and length comparisons on discrete linear arrays of dots and continuous rectangles. The results show the bilateral involvement of parietal regions around the intraparietal sulci in explicit and implicit processing of numerosity, and a right lateralized occipitoparietal network activation in length processing; numerosity and length processing both activate the right IPS and the precentral gyrus. By excluding the mandatory intrinsic spatial processing of arrays, we demonstrate that the left IPS is involved in numerosity processing only, whereas the right IPS underlies a common processing mechanism or representation of spatial and numerical magnitude. PMID- 19294654 TI - Disulfide bonds versus TrpTrp pairs in irregular beta-hairpins: NMR structure of vammin loop 3-derived peptides as a case study. AB - Structural studies on model peptides have led to a good understanding of the rules behind the formation and stability of regular beta-hairpins. To test their applicability to the successful design of irregular beta-hairpins with long loops and/or beta-bulges at the strands, we mimicked loop 3 of vammin, a 4:6 beta hairpin with a non-Gly beta-bulge. The most stabilising cross-strand pairs, disulfide bonds or/and TrpTrp pairs, were incorporated at non-hydrogen-bonded sites in peptides spanning the 69-80 region of vammin. According to NMR data, these modified peptides adopt beta-hairpin conformations as intended by design. The Trp-containing peptides reproduce even the unusual positive phi angle for the Gln residue, with the indole rings in the preferred edge-to-face orientation. For the first time the beta-hairpin-stabilising capacities of a disulfide bond and a TrpTrp pair are compared in the same model system. We found that the contribution to stability of the noncovalent indole-indole interaction is larger than that of the covalent disulfide bond, and that their combination gives rise to an even more stable beta-hairpin. PMID- 19294658 TI - Synthesis, characterization, and electro-optical properties of Zn(II) complexes with pi-conjugated terpyridine ligands. AB - A series of nine zinc(II) complexes containing substituted 4'-phenyl-2,2':6',2'' terpyridines as ligands is synthesized and fully characterized. The ground-state structures of four examples are calculated by means of DFT and their structural features are confirmed by experimental Raman spectroscopy. Special focus is placed on the degree of pi-electron delocalization between the terpyridine unit and the attached phenyl moiety. Applying Bader's quantum theory of atoms in molecules (QTAIM) and visualizing the electron-density distribution by intermolecular Deltarho plots reveals an increase in ellipticity-and therefore pi electron delocalization-for phenylvinyl-substituted derivatives compared to phenylethynyl-substituted ones. Experimentally, this is verified by spectroscopic means, because an increase in ellipticity goes along with a pronounced decrease of the HOMO-LUMO energy band gap. Overall, the lateral pi-conjugated substituents are found to strongly influence the electro-optical properties of the complexes. In solution, the color of emission can be modulated from violet to cyan (425-487 nm) and high quantum yields (Phi(PL) up to 0.60) are observed. Thin solid films of the complexes in a matrix of poly(methyl methacrylate) have been inkjet printed, and their photophysical behavior (bright emission, Phi(PL) up to 0.30) reveals their potential as new emissive materials for applications in light emitting devices. PMID- 19294660 TI - Flow-injection analysis for meloxicam based on tris(2,2'-bipyridine) ruthenium(II)-Ce(IV) chemiluminescent system. AB - It was found that meloxicam could enhance the chemiluminescence (CL) of the tris(2,2'-bipyridine) ruthenium(II)-Ce(IV) system in the medium of sulfate acid. Based on this phenomenon a new flow-injection system with chemiluminescent detection has been proposed for determination of meloxicam. Under optimum conditions, meloxicam had a good linear relationship with the CL intensity in the concentration range of 6.0 10(-4) to 1.0 microg/mL and the detection limit was 3.7 x 10(-4 )microg/mL. The proposed method was applied to detect meloxicam in tablets and a satisfactory recovery was obtained. The possible mechanism for this CL system is also discussed in this paper. PMID- 19294661 TI - Fluorescence sensing of nitric oxide in aqueous solution by triethanolamine modified CdSe quantum dots. AB - The water-soluble luminescent CdSe quantum dots were prepared by ligand exchange with triethanolamine (TEA). Oxygen can reversibly enhance the fluorescence of the synthesized quantum dots (TEA-CdSe-QDs) in aqueous solution. Nitric oxide radical (NO) can react easily with dissolved oxygen in water and was found to have a significant quenching effect on the fluorescence of the TEA-CdSe-QDs. The fluorescence responses were concentration-dependent and can be well described by the typical Stern-Volmer equation. A good linear relationship (R(2 )= 0.9963) was observed over the range 5.92 x 10(-7 )to 1.85 x 10(-5) mol/L nitric oxide. Above this concentration was a second linear region ranging from 2.12 x 10(-5) to 1.12 x 10(-4 )mol/L NO with a gentler slope. The detection limit, calculated following the 3sigma IUPAC criteria, was 3.02 x 10(-7 )mol/L. The interference effect of some common interferents such as nitrite (NO2 (-)), nitrate (NO(3) (-)), glucose and l-ascorbic acid on the detection of NO was negligible for the proposed system, demonstrating the potential utility of this probe for the detection of NO in biological systems. The possible mechanism was also discussed. PMID- 19294662 TI - Comparative sugar recovery and fermentation data following pretreatment of poplar wood by leading technologies. AB - Through a Biomass Refining Consortium for Applied Fundamentals and Innovation among Auburn University, Dartmouth College, Michigan State University, the National Renewable Energy Laboratory, Purdue University, Texas A&M University, the University of British Columbia, and the University of California at Riverside, leading pretreatment technologies based on ammonia fiber expansion, aqueous ammonia recycle, dilute sulfuric acid, lime, neutral pH, and sulfur dioxide were applied to a single source of poplar wood, and the remaining solids from each technology were hydrolyzed to sugars using the same enzymes. Identical analytical methods and a consistent material balance methodology were employed to develop comparative performance data for each combination of pretreatment and enzymes. Overall, compared to data with corn stover employed previously, the results showed that poplar was more recalcitrant to conversion to sugars and that sugar yields from the combined operations of pretreatment and enzymatic hydrolysis varied more among pretreatments. However, application of more severe pretreatment conditions gave good yields from sulfur dioxide and lime, and a recombinant yeast strain fermented the mixed stream of glucose and xylose sugars released by enzymatic hydrolysis of water washed solids from all pretreatments to ethanol with similarly high yields. An Agricultural and Industrial Advisory Board followed progress and helped steer the research to meet scientific and commercial needs. PMID- 19294663 TI - A novel inhibitory kinetic fluorimetric method for the determination of trace methomyl in environmental samples. AB - A novel inhibitory kinetic fluorimetric method for the determination of trace methomyl was proposed. It was shown that the Fenton reagent oxidized rhodamine B in acid medium which enabled the fluorescence quenching of the latter. The presence of trace methomyl clearly inhibited the reaction. Upon addition of EDTA, a good linear relationship between the inhibitory effect and the concentration of methomyl was observed, together with improved stabilization and sensitivity. Factors affecting the determination of trace methomyl were investigated systematically. Under the optimum conditions, the linear range for the determination of methomyl was 0.04-2.2 microg/mL; the detection limit and the quantification limit for methomyl were 0.011 and 0.037 microg/mL, respectively. The proposed method was applied to the determination of methomyl in four environmental soil samples, six environmental water samples and one synthetic sample; the results were compared with those determined by the HPLC method. The recoveries and the relative errors were 83.5-101.2 and 0.47-2.02%, respectively. The possible reaction mechanism has also been discussed. PMID- 19294671 TI - Metal-free organic dyes for dye-sensitized solar cells: from structure: property relationships to design rules. AB - Dye-sensitized solar cells (DSSC) have attracted considerable attention in recent years as they offer the possibility of low-cost conversion of photovoltaic energy. This Review focuses on recent advances in molecular design and technological aspects of metal-free organic dyes for applications in dye sensitized solar cells. Special attention has been paid to the design principles of these dyes and on the effect of various electrolyte systems. Cosensitization, an emerging technique to extend the absorption range, is also discussed as a way to improve the performance of the device. In addition, we report on inverted dyes for photocathodes, which constitutes a relatively new approach for the production of tandem cells. Special consideration has been paid to the correlation between the molecular structure and physical properties to their performance in DSSCs. PMID- 19294672 TI - Prognostic effect of pathologic fracture in localized osteosarcoma: a cohort/case controlled study at a single institute. AB - BACKGROUND: The negative prognostic role of pathologic fracture in osteosarcoma is not determined, as previous case-control and retrospective cohort studies have produced contradictory results. METHODS: We conducted both cohort (n = 384) and case-control (n = 111) studies on 37 pathologically fractured localized osteosarcoma of extremity. RESULTS: In cohort study, patients with a fracture showed a tendency of poorer survival, but the difference did not reach the level of significance (5-year metastasis-free survival rates; 48% for cases vs. 61% for controls; P = 0.06). A case-control study on 37 fractured and 74 control recruited from 347 patients matched for tumor size and location showed no survival difference between the cases and controls (P = 0.12). CONCLUSIONS: Reported negative prognostic effect of a pathologic fracture is likely to be due to confounding by tumor size and location. The present study suggests that the presence of a pathologic fracture has no prognostic relevance. PMID- 19294673 TI - Sodium benzoate exposure downregulates the expression of tyrosine hydroxylase and dopamine transporter in dopaminergic neurons in developing zebrafish. AB - BACKGROUND: Recent data have demonstrated that treatment with sodium benzoate (SB) leads to significant developmental defects in motor neuron axons and neuromuscular junctions in zebrafish larvae, thereby implying that SB can be neurotoxic. This study examined whether SB affects the development of dopaminergic neurons in the zebrafish brain. METHODS: Zebrafish embryos were exposed to different concentrations of SB for various durations, during which the survival rates were recorded, the expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the neurons in the ventral diencephalon were detected by in situ hybridization and immunofluorescence, and the locomotor activity of larval zebrafish was measured. RESULTS: The survival rates were significantly decreased with the increase of duration and dose of SB-treatment. Compared to untreated clutch mates (untreated controls), treatment with SB significantly downregulated expression of TH and DAT in neurons in the ventral diencephalon of 3-day post-fertilization (dpf) zebrafish embryos in a dose dependent manner. Furthermore, there was a marked decrease in locomotor activity in zebrafish larvae at 6dpf in response to SB treatment. CONCLUSIONS: The results suggest that SB exposure can cause significantly decreased survival rates of zebrafish embryos in a time- and dose-dependent manner and downregulated expression of TH and DAT in dopaminergic neurons in the zebrafish ventral diencephalon, which results in decreased locomotor activity of zebrafish larvae. This study may provide some important information for further elucidating the mechanism underlying SB-induced developmental neurotoxicity. PMID- 19294678 TI - Antibiotic prophylaxis before second-trimester genetic amniocentesis (APGA): a single-centre open randomised controlled trial. AB - OBJECTIVE: To compare procedure-related pregnancy loss after second-trimester genetic amniocentesis in women given an antibiotic prophylaxis and controls. METHODS: Prospective, open randomised controlled single-centre study between January 1999 and December 2005 at Artemisia Fetal Maternal Medical Centre. A follow-up within 4 weeks after the procedure was done.Of 36,347 eligible women, 1424 refused to participate and 34,923 were enrolled and randomised with unequal chance of selection, 21,991 were assigned to treatment group and 12,932 were assigned to the control group, and did not receive any placebo. Oral azithromycin, 500 mg per day, was administered 3 days before amniocentesis. The primary endpoint was the procedure-related pregnancy loss. The secondary endpoint was the rate of preterm premature rupture of membranes. RESULTS: The rate of abortion related to the amniocentesis was 7/21 219 women (0.03%, 95% CI 0.009 0.057) in the intervention group, and 36/12 529 (0.28%, 0.28-0.30) in controls (p = 0.0019). The rate of preterm premature rupture of membranes was 14/21 219 (0.06%, 0.031-0.101) in the intervention group, and 140/12 529 (1.12%, 0.94-1.30) in the control group (p = 0.001). CONCLUSIONS: Antibiotic prophylaxis before second-trimester amniocentesis reduced the risk of abortion and of rupture of the membranes. PMID- 19294679 TI - Effects of natalizumab, an alpha4 integrin inhibitor, on the development of Hartley guinea pigs. AB - BACKGROUND: Natalizumab is a humanized monoclonal immunoglobulin G4 antibody directed against the human alpha4 integrin subunit, disrupting interaction with its ligands. Natalizumab inhibits the interaction of alpha4 integrins with fibronectin, vascular cell adhesion molecule-1, and mucosal addressin cellular adhesion molecule-1, which are of potential importance in development. Two studies were undertaken to evaluate the effects of natalizumab on embryo/fetal development in guinea pigs. METHODS: In the first study, pregnant guinea pigs were treated with intravenous injections of 3, 10, or 30 mg/kg natalizumab or vehicle every other day from gestational day (GD) 4 to 30. In the second study, females were treated on alternate days starting at least 28 days prior to mating through GD 30. Fetal examinations and histopathologic examination of the liver, heart, thymus, spleen, and intestinal tract were performed following maternal euthanasia on GD 59-62. RESULTS: Natalizumab had no significant effect on embryo/fetal development in either study. Exposure to natalizumab during organogenesis did not result in treatment-related external, visceral, or skeletal variations or malformations or histopathologic changes. CONCLUSION: No fetotoxicity or teratogenic effects were attributable to natalizumab in these studies. PMID- 19294680 TI - Evaluation of the safety and pharmacokinetics of the multi-targeted receptor tyrosine kinase inhibitor sunitinib during embryo-fetal development in rats and rabbits. AB - BACKGROUND: Angiogenesis plays a key role in embryo-fetal development and, based on nonclinical safety data, the majority of vascular endothelial growth factor (VEGF)-targeted antiangiogenic agents used in cancer therapy are not recommended during pregnancy. We investigated the effects of sunitinib (an oral inhibitor of multiple receptor tyrosine kinases [RTKs] including VEGF-receptors) on embryo fetal development. METHODS: Presumed-pregnant Sprague-Dawley rats and New Zealand White rabbits received repeated daily oral doses of sunitinib (0-30 mg/kg/day), during the major period of organogenesis. Clinical/physical examinations were performed throughout the gestation phase, and blood samples were collected to determine systemic exposure. Necropsy (including uterine examination) was performed on all animals and fetal morphology was examined. RESULTS: The no observed-adverse-effect level was 1-5 mg/kg/day for maternal toxicity and 3 mg/kg/day for developmental toxicity in rats; 1 and 0.5 mg/kg/day, respectively, in rabbits. Embryo-fetal toxicity included decreases in the number of live fetuses and increases in the numbers of resorptions and post implantation/complete litter losses; these were observed at doses of > or =5 mg/kg/day in rats and 5 mg/kg/day in rabbits. Malformations included fetal skeletal malformations (generally thoracic/lumbar vertebral alterations) in rats and cleft lip/palate in rabbits. These developmental effects were observed at approximately 5.5- (rats) and approximately 0.3-times (rabbits) the human systemic exposure at the approved sunitinib dose (50 mg/day). CONCLUSIONS: Similar effects have been reported with the prototype monoclonal antibody bevacizumab. As is typically observed for potent inhibitors of RTKs involved in angiogenesis, sunitinib was associated with embryo-fetal developmental toxicity in rats and rabbits at clinically relevant dose levels. PMID- 19294681 TI - Digestive proteolysis organization in two closely related Tenebrionid beetles: red flour beetle (Tribolium castaneum) and confused flour beetle (Tribolium confusum). AB - The spectra of Tribolium castaneum and T. confusum larval digestive peptidases were characterized with respect to the spatial organization of protein digestion in the midgut. The pH of midgut contents in both species increased from 5.6-6.0 in the anterior to 7.0-7.5 in the posterior midgut. However, the pH optimum of the total proteolytic activity of the gut extract from either insect was pH 4.1. Approximately 80% of the total proteolytic activity was in the anterior and 20% in the posterior midgut of either insect when evaluated in buffers simulating the pH and reducing conditions characteristic for each midgut section. The general peptidase activity of gut extracts from either insect in pH 5.6 buffer was mostly due to cysteine peptidases. In the weakly alkaline conditions of the posterior midgut, the serine peptidase contribution was 31 and 41% in T. castaneum and T. confusum, respectively. A postelectrophoretic peptidase activity assay with gelatin also revealed the important contribution of cysteine peptidases in protein digestion in both Tribolium species. The use of a postelectrophoretic activity assay with p-nitroanilide substrates and specific inhibitors revealed a set of cysteine and serine endopeptidases, 8 and 10 for T. castaneum, and 7 and 9 for T. confusum, respectively. Serine peptidases included trypsin-, chymotrypsin , and elastase-like enzymes, the latter being for the first time reported in Tenebrionid insects. These data support a complex system of protein digestion in the Tribolium midgut with the fundamental role of cysteine peptidases. PMID- 19294682 TI - Amniocentesis in the second trimester and congenital talipes equinovarus in the offspring: a population-based record linkage study in Scotland. AB - BACKGROUND: To investigate whether amniocentesis in the second trimester is associated with congenital talipes equinovarus (CTEV) in the offspring. METHODS: Case-control study nested within a population-based cohort, developed through linkage of the Scottish Congenital Anomalies Linked Database with records of amniocentesis from cytogenetics laboratories, including 564,299 singleton births 1992-2001. Odds ratios and 95% confidence intervals for CTEV in the offspring (isolated, non-isolated, total) were calculated using logistic regression, adjusting for maternal age, year of birth and health board of birth. RESULTS: There was a modest positive association between total CTEV and amniocentesis at any time (OR = 1.27, 95% CI 0.99-1.65) and at >or= 15 weeks (OR = 1.25, 95%CI 0.95-1.64). The association was strongest for non-isolated CTEV (amniocentesis any time: OR = 1.68, 95%CI 1.08-2.61; amniocentesis >or= 15 weeks: OR = 1.81, 95%CI 1.16-2.83). Amniocentesis at >or= 20 weeks was associated with increased risk of total (OR = 5.87, 95% CI 3.38-10.21), non-isolated (OR = 13.17, 95% CI 6.49-26.74) and isolated CTEV (OR = 3.10, 95% CI 1.28-7.49). There were no associations in mothers aged >or= 35 years. CONCLUSIONS: The modest association observed is most likely accounted for by amniocenteses conducted because of an earlier abnormal prenatal test. Thus, second trimester amniocentesis is unlikely to contribute to the development of CTEV in the offspring. PMID- 19294683 TI - Neutral and ion thermochemistry: its present status and significance. AB - This short account outlines the sources of thermochemical data that are important for gas phase ion chemistry. It describes some of the relationships that have been identified for the empirical estimation of enthalpies among neutral molecules, free radicals, and odd and even electron ions. For neutral species, the additivity principle works well and this has been developed to cover a very wide range of structures and isomers. Ionization energies of homologous species depend inversely on molecular size, allowing estimates to be made for missing members. For ions, the effect of a group substitution (such as replacing a hydrogen atom by, e.g., a methyl or hydroxyl group) can easily be estimated, but such results are strongly dependent upon the position of the charge site, relative to that of the substitution. Special emphasis is given to the reliability of data collections and simple directions are provided as to how critically to assess and identify less-than-satisfactory values. PMID- 19294684 TI - Non-blinking semiconductor colloidal quantum dots for biology, optoelectronics and quantum optics. AB - Twinkle, twinkle: The blinking of semiconductor colloidal nanocrystals is the main inconvenience of these bright nanoemitters. There are various approaches for obtaining non-blinking nanocrystals, one of which is to grow a thick coat of CdS on the CdSe core (see picture). Applications of this method in the fields of optoelectronic devices, biologic labelling and quantum information processing are discussed.The blinking of semiconductor colloidal nanocrystals is the main inconvenience of these bright nanoemitters. For some years, research on this phenomenon has demonstrated the possibility to progress beyond this problem by suppressing this fluorescence intermittency in various ways. After a brief overview on the microscopic mechanism of blinking, we review the various approaches used to obtain non-blinking nanocrystals and discuss the commitment of this crucial improvement to applications in the fields of optoelectronic devices, biologic labelling and quantum information processing. PMID- 19294685 TI - Attosecond quantum stroboscope. AB - Electron disco: A "quantum stroboscope" for capturing the electron motion on a subfemtosecond timescale for a particular class of problems is highlighted. The picture shows a diffraction pattern caused by wave packets obtained by synchronizing attosecond UV pulses to a near-IR field and ionizing rare-gas atoms. PMID- 19294686 TI - Host-assisted guest self-assembly: enhancement of the dimerization of pyronines Y and B by gamma-cyclodextrin. AB - Buckle up! The dimerization of small fluorescent guests is strongly enhanced in presence of a cyclodextrin host. The host cavity acts like a belt to assist the self-assembly of guests (see picture). Small variations in the guest structure have significant influence on the stability and geometry of the aggregates.The role of small variations in the structural properties of host and guest molecules on the stoichiometry and strength of supramolecular associations is analyzed. Earlier we found that a change in substituents from pyronine B to pyronine Y has a dramatic effect on both the stability and the dynamics of the association of these guests with beta-cyclodextrin as host. Now we study the association between these two pyronines and a cyclodextrin with a bigger cavity (gamma-cyclodextrin) using UV/Vis absorption and fluorescence spectroscopy. The absorption spectra of the pyronines show complex variations with cyclodextrin concentration indicating that pyronine dimerization is strongly enhanced inside the cavity of the cyclodextrin. A full model is proposed and the equilibrium constants of the involved processes and the absorption and emission spectra of the different species are estimated. The equilibrium constants of the formation of complexed dimers are much higher than those for free dimerization or for the inclusion of a single guest. The gamma-cyclodextrin host acts like a belt to assist the guest self-assembly. The differences in the stability of pyronine B and pyronine Y dimers are explained on the basis of their structure and geometry. PMID- 19294687 TI - Conformational dynamics of DNA-electrophoresis on cationic membranes. AB - The conformational dynamics of DNA molecules undergoing electrophoresis on a fluid substrate-supported cationic lipid bilayer is investigated using fluorescence microscopy. At low electrophoretic velocities, drift of 2-D random coils is observed. In contrast, at velocities larger than 0.3 mum/s, the DNA molecules stretch out and assume branched configurations. The cross-over scenario is explained by the observation that cationic lipids segregate underneath the adsorbed DNA and confine the DNA to its counter charge imprint on time scales shorter than the relaxation time of the imprint. The concept of a tube-like confinement of the DNA is corroborated by the observed 1/N size dependence of the electrophoretic mobility in analogy to the biased reptation model in gels. The role of membrane defects and possible applications of membrane-based electrophoresis in microfluidic devices are discussed. PMID- 19294688 TI - Improving the sensitivity of negative controls in ancient DNA extractions. AB - Much attention has been paid on ancient DNA (aDNA) studies, and negative control was used as one of the stringent quality assurance criteria in order to detect potential contamination. However, the results of some aDNA studies showed the evidence of contamination despite their negative controls failed to do so. Using lambda DNA to mock extraneous contaminating DNA, our study showed that aDNA had a property of improving the efficiency of extraction including contaminating DNA, while negative controls had low sensitivity to detect contamination. To circumvent this problem, carrier DNA such as poly(dA) is suggested to be introduced into aDNA extraction. PMID- 19294690 TI - SELDI-TOF as a method for biomarker discovery in the urine of aristolochic-acid treated mice. AB - Aristolochic acids (AAs) present in Aristolochia plants are substances responsible for Chinese herbs nephropathy. Recently, strong indications have also been presented, which dietary poisoning with AA is responsible for endemic (Balkan) nephropathy (EN), an enigmatic renal disease that affects rural population living in some countries in Southeastern Europe. A mouse model was applied to follow the effects of two forms of AA, AAI and AAII. SDS-PAGE and SELDI-TOF mass spectrometry with normal phase chips were used to evaluate changes in the urine of treated animals. These two methods are demonstrated to be comparable. The use of SELDI-TOF MS for rapid analysis of a large number of samples and the combination of this method with nano-LC-ESI MS/MS for protein identification were demonstrated. Biomarker discovery after analysis of large cohort of EN patients will be the final aim of these investigations. PMID- 19294691 TI - The use of CE ECL with ionic liquid for the determination of drug alkaloids and applications in human urine. AB - A new approach for the determination of methylephedrine hydrochloride (ME), thebaine, codeine phosphate (CP), and acetylcodeine (AC) was established by CE ECL with ionic liquid (1-butyl-3-methylimidazolium tetrafluoroborate). The conditions for the CE separation, ECL detection, and the effect of ionic liquid were systematically investigated. Under the optimal conditions, the four analytes were well separated within 8 min using 1-butyl-3-methylimidazolium tetrafluoroborate as additive in the electrophoretic buffer. The concentration detection limits of ME, thebaine, CP, and AC were 2.1 x 10(-8), 1.4 x 10(-7), 6.3 x 10(-8), and 3.6 x 10(-8) mol/L (S/N=3), respectively. The LOQs (S/N=10) in real human urine samples were 7.6 x 10(-7) mol/L for ME, 3.6 x 10(-6) mol/L for thebaine, 6.5 x 10(-7) mol/L for CP, and 4.6 x 10(-7) mol/L for AC, respectively. The recoveries of four alkaloids at different levels in human urine samples were between 90.0 and 103.5%. The developed method was successfully applied to the determination of four drug alkaloids in human urine samples. PMID- 19294692 TI - Lipid metabolite levels of prostaglandin D2 and eicosapentaenoic acid recovered from bronchoalveolar lavage fluid correlate with lung function of chronic obstructive pulmonary disease patients and controls. AB - One-step global profiling of analyte (mRNA, protein, metabolite) biomarkers may soon replace conventional blood and histological/biopsy diagnostics technologies. It is important to establish whether the numerous blood and other body fluid derived potential novel diagnostics will be sufficiently efficacious and precise to replace, for example, imaging and functional diagnostic tests. Currently, imaging technologies and spirometry are indispensable for the diagnosis and management of chronic obstructive pulmonary disease (COPD). To validate the concept of using body fluid biomarkers in COPD and to address the question of whether biomarker levels correlate with lung function, we measured the level of a number of biologically relevant lipids and metabolites in the bronchoalveolar lavage (BAL) fluid of COPD and control subjects and examined whether these correlate with numeric parameters of lung function. Both the diagnosis and management of COPD rely on costly and labor intensive lung function tests. Thus, there is an imminent need to replace the current diagnostic approaches with simpler clinical assays. As a first step, we demonstrate proof of principle; the correlation of lipid biomarkers as measured by LC-MS with lung function. In the apparently BAL-accessible fluid compartment, the total recovered lipid metabolite amount, particularly prostaglandin D(2) and eicosapentaenoic acid show a remarkable linear correlation with lung function (R(2)>0.7). The study outcome is encouraging for the continuation of the work toward the measurement of lipid metabolite levels in more easily obtainable biological fluids such as sputum, exhaled air condensate, urine and plasma. PMID- 19294693 TI - Atorvastatin modifies the protein profile of circulating human monocytes after an acute coronary syndrome. AB - Aggressive treatment with high-dose atorvastatin reduces more effectively the incidence of cardiovascular events than moderate statin therapy. The mechanism of this benefit has not been fully elucidated. In order to know the potential effects of statin treatment on the protein expression of circulating monocytes in acute coronary syndrome (ACS) patients, a proteomic analysis of these cells was carried out by 2-DE and MS. Twenty-five patients with non-ST-elevation acute coronary syndrome (NSTEACS) were randomized, the fourth day after admission, to receive ATV 80 mg/dL (n = 14) or conventional treatment (CT) (n = 11), for two months. Blood was withdrawn at the end of the treatment, and monocytes were extracted for proteomic analysis and their protein expression patterns determined. Age, sex, total cholesterol, LDL, HDL, triglycerides, body mass index, presence of hypertension, diabetes, and smoking status were not significantly different between the two groups of patients. The expression of 20 proteins was modified by intensive ATV. Among the most relevant results stand out the normalization by intensive ATV treatment of the expression of proteins that modulate inflammation and thrombosis such as protein disulfide isomerase ER60 (PDI), Annexin I, and prohibitin, or that have other protective effects as HSP 70. Thus, this approach shed light at the molecular level of the beneficial mechanisms of anti-atherothrombotic drugs. PMID- 19294694 TI - Alterations to the protein profile of bladder carcinoma cell lines induced by plant extract MINA-05 in vitro. AB - Bladder cancer (BLCa) is a severe urological cancer of both men and women that commonly recurs and once invasive, is difficult to treat. MINA-05 (CK Life Sciences Int'l, Hong Kong) is a derivative of complex botanical extracts, shown to reduce cellular proliferation of bladder and prostate carcinomas. We tested the effects of MINA-05 against human BLCa cell sublines, B8, B8-RSP-GCK, B8-RSP LN and C3, from a transitional cell carcinoma, grade IV, to determine the molecular targets of treatment by observing the cellular protein profile. Cells were acclimatised for 48 h then treated for 72 h with concentrations of MINA-05 reflecting 1/2 IC(50), IC(50) and 2 x IC(50) (n = 3) or with vehicle, (0.5% DMSO). Dose-dependant changes in protein abundance were detected and characterised using 2-dimensional electrophoresis and MS. We identified 10 proteins that underwent changes in abundance, pI and/or molecular mass in response to treatment. MINA-05 was shown to influence proteins across numerous functional classes including cytoskeletal proteins, energy metabolism proteins, protein degradation proteins and tumour suppressors, suggesting a global impact on these cell lines. This study implies that the ability of MINA-05 to retard cellular proliferation is attributed to its ability to alter cell cycling, metabolism, protein degradation and the cancer cell environment. PMID- 19294689 TI - A new method for preparation of an etched porous joint for capillary electrophoresis and its pore-size evaluation. AB - In this work, an improved approach was developed for the construction of a hydrofluoric acid etched capillary porous joint, and the pore size of the porous joint was investigated. Several etching factors such as reagents concentration and etching temperature were optimized. Results showed that moderate temperature with low hydrofluoric acid concentration was helpful for proportional etching. Low-range protein molecular weight marker labeled with acridinium ester was used as pore-size reference for pore-size evaluation of the joint. A homemade CE with chemiluminescence detection system was used for the analysis of the acridinium ester labeled protein molecular weight marker, and the results showed that only the small molecular peptides can pass through the etched porous joint and the big size proteins cannot. The penetration of protein whose molecular mass was larger than 6500 Da was a signal of over-etching. PMID- 19294695 TI - Synergistic effect of trichostatin A and 5-aza-2'-deoxycytidine on growth inhibition of pancreatic endocrine tumour cell lines: a proteomic study. AB - Our research group recently reported that pancreatic endocrine cancer cell lines are sensitive to the HDAC inhibitor trichostatin A (TSA). In the present paper, we show that the combined treatment of pancreatic endocrine tumour cell lines with TSA and the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (DAC) determines a strong synergistic inhibition of proliferation mainly due to apoptotic cell death. Proteomic analysis demonstrates that the modulation of specific proteins correlates with the antiproliferative effect of the drugs. A schematic network clarifies the most important targets or pathways involved in pancreatic endocrine cancer growth inhibition by single or combined drug treatments, which include proteasome, mitochondrial apoptotic pathway and caspase related proteins, p53 and Ras related proteins. A comparison between the patterns of proteins regulated by TSA or DAC in endocrine and ductal pancreatic cancer cell lines is also presented. PMID- 19294696 TI - Benchmarking currently available SELDI-TOF MS preprocessing techniques. AB - SELDI protein profiling experiments can be used as a first step in studying the pathogenesis of various diseases such as cancer. There are a plethora of software packages available for doing the preprocessing of SELDI data, each with many options and written from different signal processing perspectives, offering many researchers choices they may not have the background or desire to make. Moreover, several studies have shown that mistakes in the preprocessing of the data can bias the biological interpretation of the study. For this reason, we conduct a large scale evaluation of available signal processing techniques to establish which are most effective. We use data generated from a standard, published simulation engine so that "truth" is known. We select the top algorithms by considering two logical performance metrics, and give our recommendations for research directions that are likely to be most promising. There is considerable opportunity for future contributions improving the signal processing of SELDI spectra. PMID- 19294697 TI - Proteomic analysis of low dose arsenic and ionizing radiation exposure on keratinocytes. AB - Human exposure to arsenic and ionizing radiation (IR) occur environmentally at low levels. While the human health effects of arsenic and IR have been examined separately, there is little information regarding their combined effects at doses approaching environmental levels. Arsenic toxicity may be affected by concurrent IR especially given their known individual carcinogenic actions at higher doses. We found that keratinocytes responded to either low dose arsenic and/or low dose IR exposure, resulting in differential proteomic expression based on 2-DE, immunoblotting and statistical analysis. Seven proteins were identified that passed a rigorous statistical screen for differential expression in 2-DE and also passed a strict statistical screen for follow-up immunoblotting. These included: alpha-enolase, epidermal-fatty acid binding protein, heat shock protein 27, histidine triad nucleotide-binding protein 1, lactate dehydrogenase A, protein disulfide isomerase precursor, and S100A9. Four proteins had combined effects that were different than would be expected based on the response to either individual toxicant. These data demonstrate a possible reaction to the combined insult that is substantially different from that of either separate treatment. Several proteins had different responses than what has been seen from high dose exposures, adding to the growing literature suggesting that the cellular responses to low dose exposures are distinct. PMID- 19294698 TI - Analysis of low molecular weight plasma proteins using ultrafiltration and large gel two-dimensional electrophoresis. AB - In this study, various solvent systems were applied to obtain a high and consistent recovery rate of low molecular weight plasma proteins (LMPP) from human plasma. A buffer system containing 7 M urea, 2 M thiourea, 25 mM NH(4)HCO(3) + 20% ACN (pH 8.2) produced the highest recovery rate of LMPP. To validate the recovery of cut off membrane (COM) obtained using the urea buffer system, 27 different 30 kDa COMs were used to prepare the LMPP sample which were then subjected to 1-D SDS-PAGE. Statistical analysis showed that the buffer system with COM produced a consistent the recovery of LMPP. In addition, 2-DE analysis was also conducted to determine the relative intensity of each protein spot. When molecular weight ranges over 30 kDa and under 30 kDa were evaluated, 953 and 587 protein spots were observed in the gels, respectively, resulting in a total of 1540 protein spots being resolved. Identification of the major proteins were then performed using a nano-LC/MS system comprised of an HPLC system and an ESI-quadrupole IT MS equipped with a nano-ESI source. PMID- 19294699 TI - Proteomic analysis of GTP cyclohydrolase 1 multiprotein complexes in cultured normal adult human astrocytes under both basal and cytokine-activated conditions. AB - GTP cyclohydrolase 1 (GCH1) is the rate-limiting enzyme of a metabolic pathway synthesizing tetrahydrobiopterin (BH(4)), the cofactor dimerizing and activating inducible nitric oxide synthase (NOS-2). GCH1 protein expression and enzyme activity are minimal in cultured, phenotypically stable, untreated normal adult human astrocytes (NAHA), but are strongly induced, together with NOS-2, by a mixture of three proinflammatory cytokines (IL-1beta, TNF-alpha, and IFN-gamma- the CM-trio) released by microglia under brain-damaging conditions. The resulting hyper-production of NO severely harms neurons. In this study, using MALDI-TOF/MS, PMF, Western immunoblotting (WB), and antibody microarrays we identified several proteins coimmunoprecipitating with GCH1. Under basal conditions, GCH1 was associated with various adaptor/regulator molecules involved in G-protein-coupled receptors signalling, protein serine/threonine phosphatase 2Cbeta (PP2Cbeta), and serine-threonine kinases like Ca(2+) calmodulin kinases (CaMKs), casein kinases (CKs), cAMP-dependent kinases (PKAs), and mitogen-activated protein kinases (MAPKs). Exposure to the three cytokines' mixture (CM-trio) significantly changed, within the 48-72 h required for the induction and activation of GCH1, the levels and identities of some of the 0 h-associated proteins: after 72 h CK IIalpha tended to dissociate from, whereas MAPK12 and JNK3 were strongly associated with fully active GCH1. These findings provide a first enticing glimpse into the intricate mechanisms regulating GCH1 activation by proinflammatory cytokines in NAHA, and may have therapeutic implications. PMID- 19294700 TI - Glycoproteomic analysis of WGA-bound glycoprotein biomarkers in sera from patients with lung adenocarcinoma. AB - Differential protein expression profiles in the serum samples from patients with lung adenocarcinoma may be associated with glycosylation during cancer development. In this study, we used various glycoproteomic approaches to investigate the different glycoproteomic profiles of human normal and lung adenocarcinoma serum samples and to investigate putative altered glycoprotein biomarkers. In our preliminary screening, FITC-labeled lectin staining was used for the detection of specific glycoprotein profiles. wheat germ agglutinin (WGA) lectin had the highest level of specific binding to glycoproteins in both samples. We enriched for glycoproteins in the serum samples using WGA lectin affinity and then performed co-immunoprecipitation with anti-haptoglobin and 2 DE, 2-D difference in-gel electrophoresis and MS analyses. From these analyses, we identified 39 differentially expressed proteins, including 27 up-regulated proteins and 12 down-regulated proteins. Bioinformatics tools were used to search for protein ontology, category classifications and prediction of glycosylation sites. In addition, three up-regulated glycoproteins (adiponectin, cerulolasmin and glycosylphosphatidyl-inositol-80) and two down-regulated glycoproteins (cyclin H and Fyn) that were found to be correlated with lung cancer development were validated by Western blot analysis. We suggest that these altered glycoproteins may be useful as biomarkers for lung cancer development and progression. PMID- 19294701 TI - Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis. AB - Microcolumn RPLC (microRPLC) is one of the optimum separation modes for shotgun proteomic analysis. To identify as many proteins as possible by MS/MS, the improvement on separation efficiency and peak capacity of muRPLC is indispensable. Although the increase in column length is one of the effective solutions, the preparation of a long microcolumn is rather difficult due to the high backpressure generated during the packing procedure. In our recent work, through connecting microcolumns of 5, 10, and 15 cm length via unions with minimal dead volume, long microcolumns with length up to 30 cm were obtained, with which 318 proteins were identified from proteins extracted from Escherichia coli by microRPLC-ESI MS/MS, and similar distributions of M(w) and pI were found with single and various coupled microcolumns. Furthermore, by using MS/MS with improved sensitivity, with such a serially coupled 30 cm long microcolumn, 1692 proteins were identified within 7 h from rat brain tissue, with false positive rate (FPR) <1%. All these results demonstrated that serially couple microcolumns might be of great promising to improve the separation capacity of microRPLC in shotgun proteomic analysis. PMID- 19294702 TI - Subproteome analysis of the neutrophil cytoskeleton. AB - Neutrophils play a key role in the early host-defense mechanisms due to their capacity to migrate into inflamed tissues and phagocytose microorganisms. The cytoskeleton has an essential role in these neutrophil functions, however, its composition is still poorly understood. We separately analyzed different cytoskeletal compartments: cytosolic skeleton, phagosome membrane skeleton, and plasma membrane skeleton. Using a proteomic approach, 138 nonredundant proteins were identified. Proteins not previously known to associate with the skeleton were: n-acetylglucosamine kinase, phosphoglycerate mutase 1, prohibitin, ficolin 1, phosphogluconate dehydrogenase, glucosidase, transketolase, major vault protein, valosin-containing protein, aldehyde dehydrogenase, and lung cancer related protein-8 (LCRP8). The majority of these proteins can be classified as energy metabolism enzymes. Such a finding was interesting because neutrophil energy metabolism is unusual, mainly relying on glycolysis. The enrichment of phosphoglycerate mutase in cytosolic skeleton was additionally indicated by the use of Western blotting. This is the broadest subcellular investigation to date of the neutrophil cytoskeletal proteome and the first proteomic analysis in any cell type of the phagosome skeleton. The association of metabolic enzymes with cytoskeleton is suggestive of the importance of their localized enrichment and macromolecular organization in neutrophils. PMID- 19294704 TI - 7th HUPO World Congress: the human disease glycomics/proteomics initiative (HGPI) session 17 August 2008, Amsterdam, The Netherlands. AB - The Human Disease Glycomics/Proteomics Initiative (HGPI) Session was held on August 17, 2008, at the HUPO World Congress in Amsterdam. Reports were made on the progress of the first and second analytical pilot studies to profile N- and O linked glycan structures of standard glycoproteins utilizing laboratories from around the world. In addition, recent advances in glycan structural analyses were presented, including the use of O-linked glycan libraries of standards, use of negative mode nano-LC-MS for O-linked glycan analysis, and identification of aberrant O-glycosylation of IgA1 as a cause of IgA nephropathy. A report was made of a newly discovered lectin, malectin, which appears to function in the folding/quality control of glycoproteins with N-linked glycans and may regulate several human disorders whose etiology involves protein quality control in the ER. The major glycan ligand for malectin was identified using a novel printed glycan microarray. Advances in the analysis of the genes that are associated with glycan expression and recognition--the glycotranscriptome--were described, as well as technologies to determine the relative quantitation of N- and O-linked glycans from as few as 2 x 10(6) cells. These technologies are being applied to identify potential biomarkers of stem and cancer cells. PMID- 19294703 TI - Assessment of albumin removal from an immunoaffinity spin column: critical implications for proteomic examination of the albuminome and albumin-depleted samples. AB - High abundance proteins in serum and plasma (e.g., albumin) are routinely removed during proteomic sample processing as they can mask lower abundance proteins and peptides of biological/clinical interest. A common method of albumin depletion is based on immunoaffinity capture, and many immunoaffinity devices are designed for multiple uses. In this case, it is critical that the albumin captured on the affinity matrix is stripped from the column prior to regeneration of the matrix and processing of subsequent samples, to ensure no carryover and that maximal binding sites are available for subsequent samples. The current study examines the ability of a manufacturer's protocol to remove the proteins and peptides captured by an immunoaffinity spin column. The data presented in the current work illustrate the difficulty in completely removing albumin from the immunoaffinity device, and consequently, may explain the variability and decreased efficiency shown for this device in previous studies. In summary, the current data present important considerations for the implementation of multiple-use immunoaffinity devices for processing subsequent clinical samples in a proteomic workflow. PMID- 19294705 TI - The 7th East Midlands Proteomics Workshop 12 November 2008, Nottingham, UK. AB - Over 200 scientists with a common interest in proteomic techniques and their application to fundamental biological and biomedical problems participated in the 7th East Midlands Proteomics Workshop. This annual one day meeting was held in Nottingham in November 2008 and is a joint venture of colleagues from three local Universities: The University of Nottingham, Nottingham Trent University, and Loughborough University. PMID- 19294706 TI - Chemical and structural diversity in eumelanins: unexplored bio-optoelectronic materials. AB - Eumelanins, the characteristic black, insoluble, and heterogeneous biopolymers of human skin, hair, and eyes, have intrigued and challenged generations of chemists, physicists, and biologists because of their unique structural and optoelectronic properties. Recently, the methods of organic chemistry have been combined with advanced spectroscopic and imaging techniques, theoretical calculations, and methods of condensed-matter physics to gradually force these materials to reveal their secrets. Herein we review the latest advances in the field with a view to showing how the emerging knowledge is not only helping to explain eumelanin functionality, but may also be translated into effective strategies for exploiting their properties to create a new class of biologically inspired high-tech materials. PMID- 19294707 TI - Targeting bone metastases with a bispecific anticancer and antiangiogenic polymer alendronate-taxane conjugate. AB - A polymer therapeutic designed for combination anticancer and antiangiogenic therapy inhibited the proliferation of prostate carcinoma cells and the proliferation, migration, and tube-formation of endothelial cells. The nanoconjugate was formed from an N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer, the bisphosphonate alendronate (for bone targeting), and the chemotherapy agent paclitaxel (PTX), which is cleaved by cathepsin B (see scheme). PMID- 19294708 TI - Enhanced hydrogen storage by palladium nanoparticles fabricated in a redox-active metal-organic framework. AB - Quick on the uptake: Palladium nanoparticles were fabricated simply by immersing {[Zn(3)(ntb)(2)(EtOH)(2)]4 EtOH}(n) (1) in an MeCN solution of Pd(NO(3))(2) at room temperature, without any extra reducing agent. 3 wt % PdNPs@[1](0.54+)(NO(3)(-))(0.54) significantly increase H(2) uptake capacities, both at 77 K and 1 bar and at 298 K and high pressures (see picture, red curve) compared to [Zn(3)(ntb)(2)](n) (black). ntb = 4,4',4''-nitrilotrisbenzoate. PMID- 19294709 TI - PELDOR spectroscopy distance fingerprinting of the octameric outer-membrane protein Wza from Escherichia coli. AB - Distance fingerprinting: Pulsed electron-electron double resonance spectroscopy (PELDOR) is applied to the octameric membrane protein complex Wza of E. coli. The data yielded a detailed distance fingerprint of its periplasmic region that compares favorably to the crystal structure. These results provide the foundation to study conformation changes from interaction with partner proteins. PMID- 19294711 TI - Bidirectional chemo-switching of spin state in a microporous framework. AB - The ins and outs of spin: Using the microporous coordination polymer {Fe(pz)[Pt(CN)(4)]} (1, pz=pyrazine), incorporating spin-crossover subunits, two directional magnetic chemo-switching is achieved at room temperature. In situ magnetic measurements following guest vapor injection show that most guest molecules transform 1 from the low-spin (LS) state to the high-spin (HS) state, whereas CS(2) uniquely causes the reverse HS-to-LS transition. PMID- 19294710 TI - Synthesis of orthorhombic Mo-V-Sb oxide species by assembly of pentagonal Mo6O21 polyoxometalate building blocks. AB - Mix and match: The pentagonal [Mo(6)O(21)](n-) polyoxomolybdate building block assembles with other sources of Mo, V, and Sb ions to form an orthorhombic Mo-V Sb oxide. The first single-crystal X-ray analysis of an orthorhombic Mo-V-based oxide, a promising catalyst for light alkane selective oxidation known as the "M1 phase", revealed the structure of the compound. PMID- 19294712 TI - Total synthesis of ciguatoxin. AB - Something fishy: Ciguatoxin (see structure) is one of the principal toxins involved in ciguatera poisoning and the target of a total synthesis involving the coupling of three segments. The key transformations in this synthesis feature acetylene-dicobalthexacarbonyl complexation. PMID- 19294713 TI - Lost in transcription--inhibition of RNA polymerase. AB - Form and function: The natural product myxopyronin A provides the key to understanding the inhibition of bacterial RNA polymerase and should spark new ideas for the design of new antibiotics against tuberculosis and other infectious diseases. PMID- 19294715 TI - 1,1,8,8-Tetramethyl[8](2,11)teropyrenophane: half of an aromatic belt and a segment of an (8,8) single-walled carbon nanotube. PMID- 19294714 TI - Iron-catalyzed chemoselective ortho arylation of aryl imines by directed C-H bond activation. AB - No Fe-ar: Iron catalyzes an imine-directed C-H bond activation to introduce an ortho-aryl group to an acetophenone-derived imine using a diarylzinc reagent (see scheme), whereas palladium catalyzes the conventional substitution reaction . The title reaction features mild and selective C-H bond activation in the presence of aryl bromide, chloride, or sulfonate groups, and 1,2-dichloroisobutane is essential to achieve such selectivity. PMID- 19294716 TI - Charge-conversional polyionic complex micelles-efficient nanocarriers for protein delivery into cytoplasm. AB - Special delivery! Polyionic complex (PIC) micelles that contain the charge conversional moieties citaconic amide or cis-aconitic amide were developed for cytoplasmic protein delivery. The increase of the charge density on the protein cargo helped the stability of the PIC micelles without cross-linking, and the charge-conversion in endosomes induced the dissociation of the PIC micelles to result in efficient endosomal release (see picture). PMID- 19294717 TI - A supramolecular polymer as a self-assembling polyvalent scaffold. AB - Binding bacteria: Discotic molecules self-assemble into columnar supramolecular polymers that show strong polyvalent binding to bacteria by virtue of mannose ligands attached at their periphery (orange; see picture). The reversible formation of the supramolecular polymers allows simple mixing of differently substituted monomers and the optimization of bacterial aggregation. PMID- 19294718 TI - Gas-phase synthesis and reactivity of the lithium acetate enolate anion, CH2CO2Li. AB - Aerial pingpong: The lithium acetate enolate anion, the prototypical lithium salt of an alpha-deprotonated carboxylate, was prepared in the gas phase by electrospray ionization (ESI) and collision-induced ionization (CID). Its structure, reactivity, and energetics are presented along with the results of high-level computations. PMID- 19294719 TI - A dimeric magnesium(I) compound as a facile two-center/two-electron reductant. AB - The odd couple: A dimeric magnesium(I) complex acts as a facile and selective two center/two-electron reductant towards a series of unsaturated substrates (see scheme; Ar = 2,6-iPr(2)C(6)H(3), Ad = 1-adamantyl). The novel reduced or reductively coupled products obtained from these reductions suggest that magnesium(I) compounds may find wide use in organic and organometallic syntheses. PMID- 19294720 TI - Redox economy in organic synthesis. AB - "Economy" is referred to as the thrifty and efficient use of material resources, as the principle of "minimum effort to reach a goal." More illuminating is: "the aim to portion one's forces in order to use as little as possible of them to reach a goal." Such statements certainly apply when the goal is to synthesize a complex target molecule. Redox economy then implies the use of as few redox steps as possible in the synthetic conquest of a target compound. While any sort of economy will help to streamline the effort of total synthesis, redox economy addresses a particularly weak area in present-day total synthesis. It is not enough to point out the present deficiencies, rather the purpose of this Review is to serve as a teaching tool for all practitioners of the field by giving and illustrating guidelines to increase redox economy in multistep organic synthesis. PMID- 19294721 TI - Evidence for C-Cl/C-Br...pi interactions as an important contribution to protein ligand binding affinity. AB - Attractive chlorine: Noncovalent interactions between chlorine or bromine atoms and aromatic rings in proteins open up a new method for the manipulation of molecular recognition. Substitution at distinct positions of two factor Xa inhibitors improves the free energy of binding by interaction with a tyrosine unit. The generality of this motif was underscored by multiple crystal structures as well as high-level quantum chemical calculations (see picture). PMID- 19294722 TI - Total synthesis of ouabagenin and ouabain. AB - A full account of the total synthesis of ouabagenin and ouabain is described. A highly stereocontrolled anionic cycloaddition for the rapid construction of the basic steroid skeleton is a pivotal conversion for the whole strategy. A careful study was needed to establish the order and the sequence of functional group manipulations. Specific conformational features of the ouabain skeleton allowed us to overcome a few stereochemical problems. Degradation studies on ouabain provided an ultimate proof for a key intermediate, which is used as a relay. Late stage butenolide formation and glycosidation yielded ouabain. PMID- 19294723 TI - Current synthesis of triterpene saponins. AB - Triterpene saponins are a structurally and biologically diverse class of glycoconjugates of triterpenes that are widely distributed in terrestrial plants and some marine organisms. Current synthesis of these complex glycoconjugates has been discussed, whereupon the stereocontrolled glycosylation, global protection/deprotection, and the total synthesis of four unusual triterpene saponins are highlighted. PMID- 19294724 TI - Synthesis and biological characterisation of novel N-alkyl-deoxynojirimycin alpha glucosidase inhibitors. AB - The N-alkylated deoxynojirimycin compound, N-(6'-(4''-azido-2'' nitrophenylamino)hexyl)-1-deoxynojirimycin (6) was synthesised as a potential photoaffinity probe for endoplasmic reticulum (ER) alpha-glucosidases I and II. Surprisingly this compound was a highly potent inhibitor of alpha-glucosidase I (IC(50), 17 nM) in an in vitro assay and proved equally effective at inhibiting cellular ER glucosidases, as determined by a free oligosaccharide (FOS) analysis. A modest library of compounds was synthesised to obtain structure-activity information by variation of the N-alkyl chain length and modifications to the azido-nitrophenyl group. All of these compounds failed to improve on the efficacy of compound 6, but most showed greater enzyme inhibitory potency than N-butyl deoxynojirimycin (NB-DNJ), a pharmacological agent that has been evaluated for the treatment of several viruses for which infectivity is dependent on host cell glycosylation. PMID- 19294726 TI - A mechanism for stochastic decision making by bacteria. AB - What mechanism underlies the induction of the lac operon? Expression from the lac operon is an all-or-none phenomenon. Recent work by Choi et al. combines single molecule imaging of gene expression with single-cell induction measurements to develop a stochastic model describing the critical role of single lac-repressor molecules in induction. PMID- 19294725 TI - Hydrogen-bond-assisted "gold cold fusion" for fabrication of 2D web structures. AB - Keeping their cool: Fabrication of a 2D weblike nanonetwork of gold was successfully demonstrated through a two-step procedure including complexation of gold precursors to a weblike supramolecular assembly of surfactant followed by in situ reduction of the precursors to gold. Molecular assemblies stabilized by hydrogen bonding provided a sound template, leading to the highly integrated structure of gold through room-temperature (cold) nanostructure fusion. PMID- 19294727 TI - Release of volatile aldehydes by the brown algal kelp Laminaria digitata in response to both biotic and abiotic stress. AB - With a little kelp from my friends: In response to biotic and abiotic stress, the brown algal kelp Laminaria digitata releases volatile fatty acid aldehydes under laboratory conditions and in its natural environment (red). In response to 4-HHE treatment, L. digitata releases (13S)-HOTrE (green). These results support the hypothesis that these compounds may mediate kelp responses to stress. PMID- 19294728 TI - Facile synthesis of ultrathin and single-crystalline Au nanowires. AB - Au nanowires have attracted significant interest in nanomaterials research owing to their chemical stability and high conductivity for potential nanoelectronic applications. However, previous syntheses yielded only polycrystalline Au nanowires with diameters larger than 10 nm. Very recent advances in solution phase synthesis have led to the formation of ultrathin single-crystalline Au nanowires with diameters of less than 10 nm. This Focus Review summarizes the synthesis, characterization, and transport studies of these ultrathin single crystalline Au nanowires for potential electronic device applications. PMID- 19294729 TI - Syntheses, photophysics, and application of iridium(III) phosphorescent emitters for highly efficient, long-life organic light-emitting diodes. AB - Rational design and synthesis of Ir(III) complexes (1-3) bearing two cyclometalated ligands (C--N) and one 2-(diphenylphosphino)phenolate chelate (P- O) as well as the corresponding Ir(III) derivatives (4-6) with only one (C--N) ligand and two P--O chelates are reported, where (C--NH)=phenylpyridine (ppyH), 1 phenylisoquinoline (piqH), and 4-phenylquinazoline (nazoH). Single crystal X-ray diffraction studies of 3 reveal a distorted octahedral coordination geometry, in which two nazo ligands adopt an eclipsed configuration, with the third P--O ligand located trans to the phenyl group of both nazo ligands, confirming the general skeletal pattern for 1-3. In sharp contrast, complex 4 reveals a trans disposition for the PPh2 groups, along with the phenolate groups residing opposite the unique cyclometalated ppy ligand, which is the representative structure for 4-6. These Ir(III) complexes exhibit green-to-red photoluminescence with moderate to high quantum efficiencies in the degassed fluid state and bright emission in the solid state. For 1-6, the resolved emission spectroscopy and relaxation dynamics are well rationalized by the computational approach. OLEDs fabricated using 12 wt. % of 3 doped in CBP and with BCP as hole blocking material, give bright electroluminescence with lambda(max)=628 nm and CIE(xy) coordinates (0.65, 0.34). The turn-on voltage is 3.2 V, while the current efficiency and the power efficiency reach 11.2 cd A(-1) and 4.5 lm W(-1) at 20 mA cm(-2). The maximum efficiency reaches 14.7 cd A(-1)and 6.8 lm W(-1) upon switching to TPBI as hole blocking material. For evaluating device lifespan, the tested device incorporating CuPc as a passivation layer, 3 doped in CTP as an emitting layer, and BAlq as hole blocking material, shows a remarkably long lifetime up to 36,000 h at an initial luminance of 500 cd m(-2). PMID- 19294730 TI - The Sod2 mutant mouse as a model for oxidative stress: a functional proteomics perspective. AB - Oxidative stress has been implicated in the pathogenesis of numerous human diseases and disorders, but the mechanistic basis often remains enigmatic. The Sod2 mutant mouse, which is sensitized to mitochondrial stress, is an ideal mutant model for studying the role of oxidative stress in a diverse range of complications arising from mitochondrial dysfunction and diminished antioxidant defense. To fully appreciate the widespread molecular consequences under increased oxidative stress, a systems approach utilizing proteomics is able to provide a global overview of the complex biological changes, which a targeted single biomolecular approach cannot address fully. This review focuses on the applications of mass spectrometry and functional proteomics in the Sod2 mouse. The combinatorial approach provides novel insights into the interplay of chemistry and biology, free radicals and proteins, thereby augmenting our understanding of how redox perturbations influence protein dynamics. Ultimately, this knowledge can lead to the development of free radical-targeted therapies. PMID- 19294731 TI - The American Society of Pediatric Hematology/Oncology (ASPHO) 2009 distinguished career award goes to Dr. Sharon Murphy. PMID- 19294732 TI - Strength-based assessment in clinical practice. AB - Strength-based assessment can enhance clinical clarity, improve the range of information, and provide a more complete picture of clients and their circumstances. Deficit-oriented assessment has improved the assessment and treatment of a number of disorders but, at the same time, has created a negative bias, considered strengths as clinical peripheries or by-products, tended to reduce clients to diagnostic categories, and created a power differential, which could be counterproductive to clinical efficacy. Strength-based assessment explores weaknesses as well as strengths to effectively deal with problems. We present a number of strength-based strategies for use in clinical practice. These strategies, we hope, will help clinicians to operationalize how strengths and weaknesses reverberate and contribute to a client's psychological status, which is comprehensive and guards against negative bias. PMID- 19294733 TI - Not so fast: speed effects on forelimb kinematics in cercopithecine monkeys and implications for digitigrade postures in primates. AB - Terrestrial mammals are characterized by their digitigrade limb postures, which are proposed to increase effective limb length (ELL) to achieve preferred or higher locomotor speeds more efficiently. Accordingly, digitigrade postures are associated with cursorial locomotion. Unlike most medium- to large-sized terrestrial mammals, terrestrial cercopithecine monkeys lack most cursorial adaptations, but still adopt digitigrade hand postures. This study investigates when and why terrestrial cercopithecine monkeys adopt digitigrade hand postures during quadrupedal locomotion. Three cercopithecine species (Papio anubis, Macaca mulatta, Erythrocebus patas) were videotaped moving unrestrained along a horizontal runway at a range of speeds (0.4-3.4 m/s). Three-dimensional forelimb kinematic data were recorded during forelimb support. Hand posture was measured as the angle between the metacarpal segments and the ground (MGA). As predicted, a larger MGA was correlated with a longer ELL. At slower speeds, subjects used digitigrade postures (larger MGA), however, contrary to expectations, all subjects used more palmigrade hand postures (smaller MGA) at faster speeds. Digitigrade postures at slower speeds may lower cost of transport by increasing ELL and step lengths. At higher speeds, palmigrade postures may be better suited to spread out high ground reaction forces across a larger portion of the hand thereby potentially decreasing stresses in hand bones. It is concluded that a digitigrade forelimb posture in primates is not an adaptation for high speed locomotion. Accordingly, digitigrady may have evolved for different reasons in primates compared to other mammalian lineages. PMID- 19294734 TI - Enzymatic digestion of liquid hot water pretreated hybrid poplar. AB - Liquid hot (LHW) water pretreatment (LHW) of lignocellulosic material enhances enzymatic conversion of cellulose to glucose by solubilizing hemicellulose fraction of the biomass, while leaving the cellulose more reactive and accessible to cellulase enzymes. Within the range of pretreatment conditions tested in this study, the optimized LHW pretreatment conditions for a 15% (wt/vol) slurry of hybrid poplar were found to be 200(o)C, 10 min, which resulted in the highest fermentable sugar yield with minimal formation of sugar decomposition products during the pretreatment. The LHW pretreatment solubilized 62% of hemicellulose as soluble oligomers. Hot-washing of the pretreated poplar slurry increased the efficiency of hydrolysis by doubling the yield of glucose for a given enzyme dose. The 15% (wt/vol) slurry of hybrid poplar, pretreated at the optimal conditions and hot-washed, resulted in 54% glucose yield by 15 FPU cellulase per gram glucan after 120 h. The hydrolysate contained 56 g/L glucose and 12 g/L xylose. The effect of cellulase loading on the enzymatic digestibility of the pretreated poplar is also reported. Total monomeric sugar yield (glucose and xylose) reached 67% after 72 h of hydrolysis when 40 FPU cellulase per gram glucan were used. An overall mass balance of the poplar-to-ethanol process was established based on the experimentally determined composition and hydrolysis efficiencies of the liquid hot water pretreated poplar. PMID- 19294735 TI - Infrared multiphoton dissociation in quadrupole ion traps. AB - The development of new ion activation techniques continues to be a dynamic area of scientific discovery, in part to complement the tremendous innovations in ionization methods that have allowed the mass spectrometric analysis of an enormous array of molecules. Ion activation/dissociation provides key information about ion structures, binding energies, and differentiation of isomers, as well as affording a primary means of identifying compounds in mixtures. Numerous new activation methods have emerged over the past two decades in an effort to develop alternatives to collisional activated dissociation, the gold standard for providing structurally diagnostic fragmentation patterns. Collisional activated dissociation does not always offer sufficiently high or controllable energy deposition, thus rendering it less useful for certain classes of molecules, such as large proteins or macromolecular complexes. Photodissociation is one of the most promising alternatives and is readily implemented in ion trapping and time of-flight mass spectrometers. Photodissociation generally entails using a laser to irradiate ions with UV, visible, or IR photons, thus resulting in internal energy deposition based on the number and wavelengths of the photons. The activation process can be extremely rapid and efficient, as well as having the potential for high total energy deposition. This review describes infrared multiphoton dissociation in quadrupole ion trap mass spectrometry. A comparison of photodissociation and collisional activated dissociation is covered, in addition to some of the methods to increase photodissociation efficiency. Numerous applications of IRMPD are discussed as well, including ones related to the analysis of drugs, peptides, nucleic acids, and oligosaccharides. PMID- 19294736 TI - Epigenetic silencing in non-neoplastic epithelia identifies E-cadherin (CDH1) as a target for chemoprevention of lobular neoplasia. AB - Invasive lobular carcinoma (ILC) of the breast is believed to develop from in situ lesions, atypical lobular hyperplasia (ALH), and lobular carcinoma in situ (LCIS). Down-regulation of the cell-cell adhesion protein E-cadherin is a defining feature of lobular breast cancer (LBC) and already occurs in ALH and LCIS. Apart from mutational mechanisms, epigenetic silencing of the E-cadherin gene (CDH1) is thought to be involved in E-cadherin down-regulation and has been observed at a high frequency in ILC. Whether CDH1 promoter methylation is already present in in situ lesions and thus contributes to the initiation of LBC is not established. We thus examined microdissected archived tissue from 20 LBCs by methylation-specific PCR to determine the CDH1 methylation status of lobular lesions. Nineteen of the 20 LBCs had a hypermethylated CDH1 promoter, including 13/14 ILCs and 13/13 ALHs or LCIS. Bisulphite sequencing indicated that methylation was complete within the investigated promoter fragment. Intriguingly, CDH1 methylation was likewise present in 8/8 adjacent non-neoplastic epithelia, but not in 6/6 mammary epithelia from healthy subjects. E-cadherin protein and mRNA were down-regulated in in situ lesions relative to adjacent epithelia. Together, these results indicate that CDH1 promoter methylation occurs in LBC prior to E-cadherin down-regulation and neoplastic formation. We thus propose that epigenetic silencing represents the first of the two hits required to silence both CDH1 alleles for LBC to develop. Because promoter methylation is in principle reversible, our findings suggest that chemoprevention of LBC by epigenetic drugs should be feasible. Furthermore, the presence of CDH1 methylation in pre-neoplastic epithelia suggests the existence of mammary regions with increased disease susceptibility, providing an explanation for the often multifocal presentation of LBC. PMID- 19294738 TI - Well-being therapy for generalized anxiety disorder. AB - Well-being therapy (WBT) aims to enhance psychological well-being based on Ryff's (1989) six dimensions: autonomy, personal growth, environmental mastery, purpose in life, positive relations, and self-acceptance. Previous studies have documented the efficacy of this psychotherapy in treating patients with mood and anxiety disorders and in preventing relapse in recurrent depression. We discuss and illustrate the use of WBT in generalized anxiety disorder. We present a case of a patient suffering from generalized anxiety disorder treated with cognitive behavioral therapy followed by WBT. PMID- 19294737 TI - A prognostic gene expression index in ovarian cancer - validation across different independent data sets. AB - Ovarian carcinoma has the highest mortality rate among gynaecological malignancies. In this project, we investigated the hypothesis that molecular markers are able to predict outcome of ovarian cancer independently of classical clinical predictors, and that these molecular markers can be validated using independent data sets. We applied a semi-supervised method for prediction of patient survival. Microarrays from a cohort of 80 ovarian carcinomas (TOC cohort) were used for the development of a predictive model, which was then evaluated in an entirely independent cohort of 118 carcinomas (Duke cohort). A 300-gene ovarian prognostic index (OPI) was generated and validated in a leave-one-out approach in the TOC cohort (Kaplan-Meier analysis, p = 0.0087). In a second validation step, the prognostic power of the OPI was confirmed in an independent data set (Duke cohort, p = 0.0063). In multivariate analysis, the OPI was independent of the post-operative residual tumour, the main clinico-pathological prognostic parameter with an adjusted hazard ratio of 6.4 (TOC cohort, CI 1.8 23.5, p = 0.0049) and 1.9 (Duke cohort, CI 1.2-3.0, p = 0.0068). We constructed a combined score of molecular data (OPI) and clinical parameters (residual tumour), which was able to define patient groups with highly significant differences in survival. The integrated analysis of gene expression data as well as residual tumour can be used for optimized assessment of the prognosis of platinum-taxol treated ovarian cancer. As traditional treatment options are limited, this analysis may be able to optimize clinical management and to identify those patients who would be candidates for new therapeutic strategies. PMID- 19294739 TI - Religious coping among Jews: development and initial validation of the JCOPE. AB - Numerous studies have underscored the importance of religious coping in psychological health and illness; however, the majority of research in this area has been conducted with Christian samples and knowledge about other religious groups is lacking. Although recent investigations have developed scales to measure religious coping among Hindus and Muslims, the potential for future research in Jewish populations remains limited as no measures of religious coping have been validated in the general Jewish community. This two-part study reports on the development and validation of the 16-item Jewish Religious Coping Scale (JCOPE). In Study 1, an exploratory factor analysis identified two factors reflecting positive and negative religious coping strategies, and the concurrent validity for the measure was evaluated by examining correlations with indices of Jewish beliefs and practices. In Study 2, a confirmatory factor analysis (CFA) verified the JCOPE's 2-factor structure, and the scale's incremental validity was evaluated by examining Jewish religious coping as a predictor of psychological distress over and above significant covariates. Results suggest that the JCOPE has good psychometric properties, and that religious coping is a significant predictor of psychological distress among Jews. PMID- 19294740 TI - Prediction of ice content in biological model solutions when frozen under high pressure. AB - High pressure is, at least, as effective as cryoprotective agents (CPAs) and are used for decreasing both homogenous nucleation and freezing temperatures. This fact gives rise to a great variety of possible cryopreservation processes under high pressure. They have not been optimized yet, since they are relatively recent and are mainly based on the pressure-temperature phase diagram of pure water. Very few phase diagrams of biological material are available under pressure. This is owing to the lack of suitable equipment and to the difficulties encountered in carrying out the measurements. Different aqueous solutions of salt and CPAs as biological models are studied in the range of 0 degrees C down to -35 degrees C, 0.1 up to 250 MPa, and 0-20% w/w total solute concentration. The phase transition curves of glycerol and of sodium chloride with either glycerol or sucrose in aqueous solutions are determined in a high hydrostatic pressure vessel. The experimental phase diagrams of binary solutions were well described by a third degree polynomial equation. It was also shown that Robinson and Stokes' equation at high pressure succeeds in predicting the phase diagrams of both binary and ternary solutions. The solute cryoconcentration and the ice content were calculated as a function of temperature and pressure conditions during the freezing of a binary solution. This information should provide a basis upon which high-pressure cryopreservation processes may be performed and the damages derived from ice formation evaluated. PMID- 19294741 TI - Genetic continuity after the collapse of the Wari empire: mitochondrial DNA profiles from Wari and post-Wari populations in the ancient Andes. AB - The Wari empire flourished in the central, highland Peruvian Andes from AD 600 1000, and although the events that led to its demise are unknown, archaeological evidence indicates that Wari control waned at the end of the first millennium. Here, we test the hypothesis that, despite the major shift in social and political organization at the fall of the Wari empire, the mitochondrial DNA (mtDNA) composition of populations from the Ayacucho Basin, the former imperial heartland of the empire, remained essentially unchanged. Results show that mtDNA haplogroup frequencies among the Wari and post-Wari groups differ, but the difference is not statistically significant (chi2 = 5.886, df = 3, P = 0.1172). This is the first study in the Andes to use haplotypic data to evaluate the observed genetic distance between two temporally distinct prehispanic populations (F(ST) = 0.029) against modeled expectations of four possible evolutionary scenarios. None of these simulations allowed the rejection of continuity. In total, at both the haplogroup and haplotype levels these data do not allow us to reject the hypothesis that post-Wari individuals sampled in this study are the maternal descendants of those sampled from the Wari era site of Conchopata. However, genetic homogeneity in the mitochondrial gene pool, as seen in the late prehispanic southern Andes, may also characterize our study region. But, prior to this research, this was unknown. If our new data show mtDNA homogeneity, then this could limit the detection of female migration if, in fact, it occurred. Nonetheless, the novel mtDNA data presented here currently do not support the hypothesis that there was an influx of genetically distinct females into the former Wari heartland after the Wari collapse. PMID- 19294742 TI - Broken ribs: paleopathological analysis of costal fractures in the human identified skeletal collection from the Museu Bocage, Lisbon, Portugal (late 19th to middle 20th centuries). AB - Although rarely reported in the anthropological literature, rib fractures are commonly found during the analysis of human skeletal remains of past and modern populations. This lack of published data precludes comparison between studies and restricts an accurate understanding either of the mechanisms involved in thoracic injuries or their impact on past societies. The present study aimed: 1) to report rib fracture prevalence in 197 individuals, 109 males, and 88 females, with ages at death ranging from 13 to 88 years old, from the Human Identified Skeletal Collection, Museu Bocage, Portugal (late 19th-middle 20th centuries); 2) to test the hypothesis that a higher prevalence of rib stress fractures existed in the 133 individuals who died from respiratory diseases, in a period before antibiotics. The macroscopic analysis revealed 23.9% (n = 47) of individuals with broken ribs. 2.6% (n = 124) out of 4,726 ribs observed were affected. Males presented more rib fractures, and a significantly higher prevalence was noted for older individuals. Fractures were more frequently unilateral (n = 34), left sided (n = 19) and mainly located on the shaft of ribs from the middle thoracic wall. Nineteen individuals presented adjacent fractured ribs. Individuals who died from pulmonary diseases were not preferentially affected. However, a higher mean rate of fractures was found in those who died from pneumonia, a scenario still common nowadays. Since rib involvement in chest wall injury and its related outcomes are important issues both for paleopathology and forensic anthropology, further investigations are warranted. PMID- 19294743 TI - Dental caries in a Portuguese identified skeletal sample from the late 19th and early 20th centuries. AB - Dental caries was investigated in 600 adult dentitions belonging to the identified osteological collections of the Museum of Anthropology, University of Coimbra, Portugal (late 19th/early 20th centuries). The main advantage of this sample compared to an archaeological source is the presence of known demographic parameters such as age, sex, and occupation. The aim of this study is to investigate the issues involved in comparing caries data derived from archaeological death assemblages with statistics compiled from clinical studies of the living. When only the upper dentition was considered, higher rates were observed in females than in males. No differences were found between sexes for lower teeth. In both sexes, both the percentage of carious teeth and the severity of lesions were found to increase with age, demonstrating that caries activity continued throughout life. The slight decrease observed for the age group 70-79 years is probably due to the increased antemortem tooth loss in the elderly. Caries was most common at contact areas (32.9%) and rarest at smooth crown surfaces (6.5%). Root surface caries was graphed in relation to the exposure of roots, and it was confirmed that the degree of root exposure was not strongly related to the frequency of carious lesions on the exposed root surface, although both increased with age. Molars were attacked more frequently by caries as a whole than premolars, canines or incisors. The results are similar to studies of recent living populations with a limited access to professional dental care. PMID- 19294744 TI - The type specimen (LB1) of Homo floresiensis did not have Laron syndrome. AB - The type specimen (LB1) of Homo floresiensis has been hypothesized to be a pathological human afflicted with Laron Syndrome (LS), a type of primary growth hormone insensitivity (Hershkovitz et al.: Am J Phys Anthropol 134 [2007] 198 208). Comparing measurements, photographs and three-dimensional, computed tomography reconstructions of LB1 with data and diagnoses from the literature on LS, we critically evaluate numerous skull and postcranial traits that Hershkovitz et al. identified as being shared by LB1 and patients with LS. The statements regarding most of these traits are new to the clinical literature and lack quantitative support. LB1 and patients with LS differ markedly in the size and shape of the cranium; thickness and pneumatization of cranial bones; morphology of the face, mandible, teeth, and chin; form of the shoulder, wrist, and pelvis; and general body proportions including relative foot size. Claims that patients with LS are similar to LB1 in displaying protracted scapulae, short clavicles, low degrees of humeral torsion, flaring ilia, and curved tibiae are not supported by data or corroborating images. Some points of similarity (e.g., femoral neck shaft angle, femoral bicondylar angle, and estimated stature) can be found in other hominins, and cannot be considered diagnostic. From our review and analysis, we conclude that LB1 did not suffer from LS. PMID- 19294745 TI - Positive interventions in clinical practice. AB - Mainstream psychotherapy has made huge strides in treating symptoms and disorders, but it has largely overlooked happiness as a therapeutic goal despite frequently hearing from clients, "Doctor, I want to be happy." This issue of Journal of Clinical Psychology: In Session describes a number of positive interventions for specific clinical problems, such as depression, anxiety, schizophrenia, loss, grief, and relationship distress. Although the name may suggest it, positive interventions do not imply that rest of psychotherapies are negative. Neither are negatives denied nor minimized. Distinct from self-help recipes proffering instant changes, positive psychology interventions refer to systematic approaches to overcome challenges by using clients' strengths and assets. A hybrid psychotherapy-coaching model and strength-based assessment can ask a client "What is right with you?" All articles are supplemented with rich case illustrations. PMID- 19294746 TI - Growth consulting: practical methods of facilitating growth through loss and adversity. AB - The way that people lead themselves and others through crisis, loss, and adversity impacts their future and the future of those around them. Traditionally, mental health professionals have emphasized the negative consequences of crisis and loss. I present a story of how the simple act of holding the door for others on September 11th created a foundation for positive interventions. I offer the OTHERS(S) model and the process of growth consulting as practical means of creating growth as a result of loss or adversity. A case illustration and examples of clinical methods to facilitate Growth Through Loss and Adversity (GTLA) are provided. PMID- 19294747 TI - Personal coaching as a positive intervention. AB - Personal coaching is a relatively new and unlicensed profession aimed at helping functioning individuals set and achieve goals, overcome obstacles, and maintain motivation. Coaching is increasingly merging with psychology as evidenced by new journals, academic programs, and research symposia. Although coaching has traditionally been used with non-clinical populations, it can be highly relevant to psychotherapy. Clinicians who develop a hybrid psychotherapy-coaching practice might be better protected from occupational stress and burnout. In addition, a number of coaching interventions and assessments might translate well to therapy and help clinicians innovate their practice. The synergy between personal coaching and psychotherapy are illustrated with a case example. PMID- 19294749 TI - The effect of continuous culture on the growth and structure of tissue-engineered cartilage. AB - The use of bioreactors for cartilage tissue engineering has become increasingly important as traditional batch-fed culture is not optimal for in vitro tissue growth. Most tissue engineering bioreactors rely on convection as the primary means to provide mass transfer; however, convective transport can also impart potentially unwanted and/or uncontrollable mechanical stimuli to the cells resident in the construct. The reliance on diffusive transport may not necessarily be ineffectual as previous studies have observed improved cartilaginous tissue growth when the constructs were cultured in elevated volumes of media. In this study, to approximate an infinite reservoir of media, we investigated the effect of continuous culture on cartilaginous tissue growth in vitro. Isolated bovine articular chondrocytes were seeded in high density, 3D culture on Millicell filters. After two weeks of preculture, the constructs were cultivated with or without continuous media flow (5-10 microL/min) for a period of one week. Tissue engineered cartilage constructs grown under continuous media flow significantly accumulated more collagen and proteoglycans (increased by 50 70%). These changes were similar in magnitude to the reported effect of through thickness perfusion without the need for large volumetric flow rates (5 10microL/min as opposed to 240-800 microL/min). Additionally, tissues grown in the reactor displayed some evidence of the stratified morphology of native cartilage as well as containing stores of intracellular glycogen. Future studies will investigate the effect of long-term continuous culture in terms of extracellular matrix accumulation and subsequent changes in mechanical function. PMID- 19294752 TI - Expression of functional Candida antarctica lipase B in a cell-free protein synthesis system derived from Escherichia coli. AB - This article reports the cell-free expression of functional Lipase B from Candida antarctica (CalB) in an Escherichia coli extract. Although most of the cell-free synthesized CalB was insoluble under conventional reaction conditions, the combined use of molecular chaperones led to the soluble expression of CalB. In addition, the functional enzyme was generated by applying the optimal redox potential. When examined using p-nitrophenyl palmitate as a substrate, the specific activity of the cell-free synthesized CalB was higher than that of the reference protein produced in Pichia pastoris. These results highlight the potential of cell-free protein synthesis technology as a powerful platform for the rapid expression, screening and analysis of industrially important enzymes. PMID- 19294753 TI - Somatostatin and gentamicin-induced auditory hair cell loss. AB - OBJECTIVE/HYPOTHESIS: Hair cells of the mammalian auditory system do not regenerate, and therefore their loss leads to irreversible hearing loss. Aminoglycosides, among other substances, can irreversibly damage hair cells. Somatostatin, a peptide with hormone/neurotransmitter properties, has neuroprotective effects by binding to its receptor. In this study, we tested whether somatostatin can protect hair cells from gentamicin-induced damage in vitro. STUDY DESIGN: This study confirmed the expression of somatostatin receptor mRNA within the cochlea and analyzed the effect of somatostatin on gentamicin induced hair cell damage and death in vitro. METHODS: Expression of somatostatin receptor mRNA in the rat cochlea was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). Protection of auditory hair cells from gentamicin was tested using two different concentrations (1 microM and 5 microM, respectively) of somatostatin. RESULTS: We detected somatostatin receptor-1 and -2 mRNA and in the organ of Corti (OC), spiral ganglion, and stria vascularis by RT-PCR. Moreover, we could see significantly less hair cell loss in the OCs that were pretreated with either 1 microM or 5 microM of somatostatin as compared with samples treated with gentamicin alone. CONCLUSIONS: Decreased hair cell loss in somatostatin-treated samples that had been exposed to gentamicin provides evidence for a protective effect of somatostatin in aminoglycoside-induced hair cell death in vitro. PMID- 19294754 TI - Prenatal diagnosis of hemoglobin Bart's disease caused by co-inheritance of two different alpha 0-thalassemia defects in China. PMID- 19294755 TI - Attitudes of Chinese couples in Hong Kong regarding using preimplantation genetic diagnosis (PGD) and human leukocyte antigens (HLA) typing to conceive a 'Saviour Child'. AB - OBJECTIVES: To investigate Hong Kong couples' attitudes and concerns about using preimplantation genetic diagnosis (PGD) and human leukocyte antigens (HLA) typing to conceive a disease-free and tissue-compatible 'Saviour Child (SC)' to save an afflicted sibling. METHODS: Two cohorts of Chinese couples, one with natural pregnancies and the other receiving in vitro fertilization (IVF) services, were studied using a structured questionnaire. RESULTS: Although most couples believed that embryos possess moral rights, they considered it acceptable to reproduce a donor child if it was safe for the child, and tissue transplantation was the only available treatment for the sick sibling. Most couples believed that the donor child would not suffer adverse physical or psychological effects but rather would gain positive psychological benefits, and opined that parents using PGD/HLA typing suffer sacrificially for their children. In response to one specific question, one-third of the couples agreed to use the donor child as a lifetime organ donor and supported the use of PGD for non-medical gender selection. One quarter were willing to reject PGD/HLA-typing because of its potential for non medical genetic enhancement. CONCLUSION: Probably influenced by the Chinese tradition of strong familism, couples in Hong Kong generally show positive attitudes towards using PGD/HLA-typing to reproduce a 'SC' to save a sibling affected with life-threatening diseases amenable to treatment with genetically compatible tissue. PMID- 19294756 TI - Methodological reporting of randomized clinical trials in major gastroenterology and hepatology journals in 2006. AB - To determine the current quality of reporting of randomized clinical trials (RCTs) in the field of gastroenterology and hepatology, we evaluated the methodological reporting of RCTs in six major gastroenterology and hepatology journals. The methodological quality, including generation of the allocation sequence, allocation concealment, double-blinding, and sample size calculation; number of patients; disease area; and funding source was also retrieved from each trial, and the relevant trials were identified by searching MEDLINE in 2006 using a highly sensitive search strategy. The status of reporting the methodological quality of RCTs was descriptively reported. One hundred five trials were included in the final analysis; of these, 81% (85/105) reported adequate generation of the allocation sequence, 61% (64/105) reported adequate allocation concealment, 51% (54/105) were double-blind, and 75% (79/105) reported adequate sample size calculation. The reported methodological quality greatly improved when compared with historical cohorts. CONCLUSION: This study shows that there was substantial improvement in the reported methodological quality in the major gastroenterology and hepatology journals, but this quality can be further improved. PMID- 19294757 TI - Post-pyloric feeding. AB - Postpyloric feeding is an important and promising alternative to parenteral nutrition. The indications for this kind of feeding are increasing and include a variety of clinical conditions, such as gastroparesis, acute pancreatitis, gastric outlet stenosis, hyperemesis (including gravida), recurrent aspiration, tracheoesophageal fistula and stenosis in gastroenterostomy. This review discusses the differences between pre- and postpyloric feeding, indications and contraindications, advantages and disadvantages, and provides an overview of the techniques of placement of various postpyloric devices. PMID- 19294758 TI - Imaging of liver cancer. AB - Improvements in imaging technology allow exploitation of the dual blood supply of the liver to aid in the identification and characterisation of both malignant and benign liver lesions. Imaging techniques available include contrast enhanced ultrasound, computed tomography and magnetic resonance imaging. This review discusses the application of several imaging techniques in the diagnosis and staging of both hepatocellular carcinoma and cholangiocarcinoma and outlines certain characteristics of benign liver lesions. The advantages of each imaging technique are highlighted, while underscoring the potential pitfalls and limitations of each imaging modality. PMID- 19294760 TI - Long-term natural history of Crohn's disease. AB - Crohn's disease is a chronic inflammatory granulomatous process that usually involves different sites in the intestinal tract. Genetic and environmental factors are thought to play a role in its etiology and pathogenesis. The disorder has a heterogeneous clinical expression and data from tertiary care settings have documented its female predominance, occasional familial nature, and high rate of stricture formation and penetrating disease. It may appear from early childhood to late adulthood, although over 80% are currently diagnosed before age 40 years, usually with terminal ileal and colonic involvement. Several studies have now shown differences in phenotypic clinical expression depending on the initial age at diagnosis, with pediatric-onset disease being more severe and more extensive with more involvement of the upper gastrointestinal tract compared to adult-onset disease. In addition, long-term studies from these tertiary care settings have documented that the disorder may evolve with time into a more complex disease with stricture formation and penetrating disease complications (i.e. fistula and abscess). Although prolonged remission with no evidence of inflammatory disease may occur, discrete periods of symptomatic and active granulomatous inflammatory disease may re-appear over many decades. Long-term studies on the natural history have also suggested that discrete events (or agents) may precipitate this granulomatous inflammatory process. PMID- 19294761 TI - Sonography of the small intestine. AB - In the last two decades, there has been substantial development in the diagnostic possibilities for examining the small intestine. Compared with computerized tomography, magnetic resonance imaging, capsule endoscopy and double-balloon endoscopy, ultrasonography has the advantage of being cheap, portable, flexible and user- and patient-friendly, while at the same time providing the clinician with image data of high temporal and spatial resolution. The method has limitations with penetration in obesity and with intestinal air impairing image quality. The flexibility ultrasonography offers the examiner also implies that a systematic approach during scanning is needed. This paper reviews the basic scanning techniques and new modalities such as contrast-enhanced ultrasound, elastography, strain rate imaging, hydrosonography, allergosonography, endoscopic sonography and nutritional imaging, and the literature on disease-specific findings in the small intestine. Some of these methods have shown clinical benefit, while others are under research and development to establish their role in the diagnostic repertoire. However, along with improved overall image quality of new ultrasound scanners, these methods have enabled more anatomical and physiological changes in the small intestine to be observed. Accordingly, ultrasound of the small intestine is an attractive clinical tool to study patients with a range of diseases. PMID- 19294759 TI - Diagnosis of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the commonest cancers worldwide, particularly in parts of the developing world, and is increasing in incidence. This article reviews the current modalities employed for the diagnosis of HCC, including serum markers, radiological techniques and histological evaluation, and summarises international guidelines for the diagnostic approach to HCC. PMID- 19294762 TI - Impact of mass screening for gluten-sensitive enteropathy in working population. AB - AIM: To assess: (1) frequency and clinical relevance of gluten sensitive enteropathy (GSE) detected by serology in a mass screening program; (2) sensitivity of antitransglutaminase (tTGA) and antiendomysium antibodies (EmA); and (3) adherence to gluten-free diet (GFD) and follow-up. METHODS: One thousand, eight hundred and sixty-eight subjects recruited from an occupational health department underwent analysis for tTGA and EmA and, if positive, duodenal biopsy, DQ2/DQ8 genotyping, clinical feature recording, blood tests, and densitometry were performed. Since > 98% of individuals had tTGA < 2 U/mL, this value was established as the cut-off limit of normality and was considered positive when confirmed twice in the same sample. Adherence to a GFD and follow up were registered. RESULTS: Twenty-six (1.39%) subjects had positive tTGA and/or EmA, and 21 underwent biopsy: six Marsh III (one IIIa, four IIIb, one IIIc), nine Marsh I and six Marsh 0 (frequency of GSE 1:125). The sensitivity of EmA for GSE was 46.6% (11.1% for Marsh I, 100% for Marsh III), while for tTGA, it was 93.3% (88.8% for Marsh I, 100% for Marsh III). All 15 patients with abnormal histology had clinical features related to GSE. Marsh I and III subjects had more abdominal pain than Marsh 0 (P = 0.029), and a similar trend was observed for distension and diarrhea. No differences in the percentage of osteopenia were found between Marsh I and III (P = 0.608). Adherence to follow-up was 69.2%. Of 15 GSE patients, 66.7% followed a GFD with 80% responding to it. CONCLUSION: GSE in the general population is frequent and clinically relevant, irrespective of histological severity. tTGA is the marker of choice. Mass screening programs are useful in identifying patients who can benefit from GFD and follow-up. PMID- 19294763 TI - Antioxidant activity of chito-oligosaccharides on pancreatic islet cells in streptozotocin-induced diabetes in rats. AB - AIM: To investigate the antioxidant activity of chito-oligosaccharides (COSs) on pancreatic islet cells in diabetic rats induced by streptozotocin. METHODS: The antioxidant effect of COSs on pancreatic islet cells was detected under optical microscopy and with colorimetric assay and gel electrophoresis. The activities of glutathione peroxidase and superoxide dismutase, total antioxidant capacity, and content of malondialdehyde in serum and tissue slices of pancreas were examined after 60 d to determine the effect of COSs in streptozotocin-induced diabetes in rats. RESULTS: COSs can prohibit the apoptosis of pancreatic islet cells. All concentrations of COSs can improve the capability of total antioxidant capacity and activity of superoxide dismutase and decrease the content of malondialdehyde drastically. Morphological investigation in the pancreas showed that COSs have resulted in the reduction of islets, loss of pancreatic cells, and nuclear pyknosis of pancreatic cells. CONCLUSION: COSs possess various biological activities and can be used in the treatment of diabetes mellitus. PMID- 19294764 TI - Curcumin suppresses PPARdelta expression and related genes in HT-29 cells. AB - AIM: To investigate the effects of curcumin on the expression of peroxisome proliferator-activated receptordelta (PPARdelta) and related genes in HT-29 cells. METHODS: HT-29 cells were treated with curcumin (0-80 micromol/L) for 24 h. The effects of curcumin on the morphology of HT-29 cells were studied by Hoechst 33342 staining. The activity of caspase-3 was determined using DEVD-pNA as substrate. The levels of peroxisome PPARdelta, 14-3-3epsilon and vascular endothelial growth factor (VEGF) in HT-29 cells were determined by Western blotting analysis and their mRNA expression was determined by real-time quantitative RT-PCR. RESULTS: Treatment with 10-80 micromol/L curcumin induced typical features of apoptosis and activated the caspase-3 in HT-29 cells. The expression of PPARdelta, 14-3-3epsilon and VEGF was reduced and the activity of beta-catenin/Tcf-4 signaling was inhibited by curcumin treatment. CONCLUSION: Curcumin can induce apoptosis of HT-29 cells and down-regulate the expression of PPARdelta, 14-3-3epsilon and VEGF in HT-29. PMID- 19294766 TI - Ex-vivo evaluation of gene therapy vectors in human pancreatic (cancer) tissue slices. AB - AIM: To culture human pancreatic tissue obtained from small resection specimens as a pre-clinical model for examining virus-host interactions. METHODS: Human pancreatic tissue samples (malignant and normal) were obtained from surgical specimens and processed immediately to tissue slices. Tissue slices were cultured ex vivo for 1-6 d in an incubator using 95% O(2). Slices were subsequently analyzed for viability and morphology. In addition the slices were incubated with different viral vectors expressing the reporter genes GFP or DsRed. Expression of these reporter genes was measured at 72 h after infection. RESULTS: With the Krumdieck tissue slicer, uniform slices could be generated from pancreatic tissue but only upon embedding the tissue in 3% low melting agarose. Immunohistological examination showed the presence of all pancreatic cell types. Pancreatic normal and cancer tissue slices could be cultured for up to 6 d, while retaining viability and a moderate to good morphology. Reporter gene expression indicated that the slices could be infected and transduced efficiently by adenoviral vectors and by adeno associated viral vectors, whereas transduction with lentiviral vectors was limited. For the adenoviral vector, the transduction seemed limited to the peripheral layers of the explants. CONCLUSION: The presented system allows reproducible processing of minimal amounts of pancreatic tissue into slices uniform in size, suitable for pre-clinical evaluation of gene therapy vectors. PMID- 19294765 TI - Management of pancreaticobiliary disease using a new intra-ductal endoscope: the Texas experience. AB - AIM: To evaluate a new single-operator mini-endoscope, Spyglass, for its performance, feasibility and safety in the management of pancreaticobiliary disease. METHODS: In a multicenter retrospective analysis of patients undergoing intraductal endoscopy, we evaluated 128 patients (71 men, mean age 57.6 years). Indications were therapeutic (TX) in 72 (56%) and diagnostic (DX) in 56 (44%). RESULTS: Peroral endoscopy was performed in 121 and percutaneous in seven. TX indications included CBD stones in 41, PD stones in six, and biliary strictures in 25. DX indications included abnormal LFT's in 15, abnormal imaging in 38 and cholangiocarcinoma staging in three. Visualization of the stone(s) was considered good in 31, fair in six, and poor in four. Advancement of the electrohydraulic lithotripsy probe was not possible in three patients and proper targeting of the lesion was partial in four patients. A holmium laser was used successfully in three patients. Ductal clearance was achieved in 37 patients after one procedure and in four patients after two procedures. Diagnosis of biliary strictures was modified in 20/29 and confirmed to be malignant in 10/23. Of the modified patients, no diagnosis was available in 17. Spyglass demonstrated malignancy in 8/17 and non-malignancy in nine. Suspected pathology by imaging studies and abnormal LFT's was modified in 43/63 (66%). Staging of cholangiocarcinoma demonstrated multicentric cholangiocarcinoma in 2/3. There was no morbidity associated with the use of Spyglass. CONCLUSION: Spyglass Spyscope is a first generation, single operator miniature endoscope that can evaluate and treat various biliary and pancreatic tract diseases. PMID- 19294767 TI - Investigation and prediction of enteral nutrition problems after percutaneous endoscopic gastrostomy. AB - AIM: To investigate and predict enteral nutrition problems after percutaneous endoscopic gastrostomy (PEG). METHODS: We retrospectively analyzed data for 252 out of 285 patients who underwent PEG at our hospital from 1999 to 2008. Enteral nutrition problems after PEG were defined as: (1) patients who required > or = 1 mo after surgery to switch to complete enteral nutrition, or who required additional parenteral alimentation continuously; or (2) patients who abandoned switching to enteral nutrition using the gastrostoma and employed other nutritional methods. We attempted to identify the predictors of problem cases by using a logistic regression analysis that examined the patients' backgrounds and the specific causes that led to their problems. RESULTS: Mean age of the patients was 75 years, and in general, their body weight was low and their overall condition was markedly poor. Blood testing revealed that patients tended to be anemic and malnourished. A total of 44 patients (17.5%) were diagnosed as having enteral nutrition problems after PEG. Major causes of the problems included pneumonia, acute enterocolitis (often Clostridium difficile-related), paralytic ileus and biliary tract infection. A multivariate analysis identified the following independent predictors for problem cases: (1) enteral nutrition before gastrectomy (a risk reduction factor); (2) presence of esophageal hiatal hernia; (3) past history of paralytic ileus; and (4) presence of chronic renal dysfunction. CONCLUSION: Enteral nutrition problems after PEG occurred at a comparatively high rate. Patient background analysis elucidated four predictive factors for the problem cases. PMID- 19294768 TI - Progression of diethylnitrosamine-induced hepatic carcinogenesis in carnitine depleted rats. AB - AIM: To investigate whether carnitine deficiency is a risk factor during the development of diethylnitrosamine (DENA)-induced hepatic carcinogenesis. METHODS: A total of 60 male Wistar albino rats were divided into six groups with 10 animals in each group. Rats in group 1 (control group) received a single intraperitoneal (i.p.) injection of normal saline. Animals in group 2 (carnitine supplemented group) were given L-carnitine (200 mg/kg per day) in drinking water for 8 wk. Animals in group 3 (carnitine-depleted group) were given D-carnitine (200 mg/kg per day) and mildronate (200 mg/kg per day) in drinking water for 8 wk. Rats in group 4 (DENA group) were injected with a single dose of DENA (200 mg/kg, i.p.) and 2 wk later received a single dose of carbon tetrachloride (2 mL/kg) by gavage as 1:1 dilution in corn oil. Animals in group 5 (DENA-carnitine depleted group) received the same treatment as group 3 and group 4. Rats in group 6 (DENA-carnitine supplemented group) received the same treatment as group 2 and group 4. RESULTS: Administration of DENA resulted in a significant increase in alanine transaminase (ALT), gamma-glutamyl transferase (G-GT), alkaline phosphatase (ALP), total bilirubin, thiobarbituric acid reactive substances (TBARS) and total nitrate/nitrite (NOx) and a significant decrease in reduced glutathione (GSH), glutathione peroxidase (GSHPx), catalase (CAT) and total carnitine content in liver tissues. In the carnitine-depleted rat model, DENA induced a dramatic increase in serum ALT, G-GT, ALP and total bilirubin, as well as a progressive reduction in total carnitine content in liver tissues. Interestingly, L-carnitine supplementation resulted in a complete reversal of the increase in liver enzymes, TBARS and NOx, and a decrease in total carnitine, GSH, GSHPx, and CAT induced by DENA, compared with the control values. Histopathological examination of liver tissues confirmed the biochemical data, where L-carnitine prevented DENA-induced hepatic carcinogenesis while D-carnitine mildronate aggravated DENA-induced hepatic damage. CONCLUSION: Data from this study suggest for the first time that: (1) carnitine deficiency is a risk factor and should be viewed as a mechanism in DENA-induced hepatic carcinogenesis; (2) oxidative stress plays an important role but is not the only cause of DENA induced hepatic carcinogenesis; and (3) long-term L-carnitine supplementation prevents the development of DENA-induced liver cancer. PMID- 19294770 TI - Anal metastasis from recurrent breast lobular carcinoma: a case report. AB - We report a case of isolated gastrointestinal metastasis from breast lobular carcinoma, which mimicked primary anal cancer. In July 2000, an 88-year-old woman presented with infiltrating lobular cancer (pT1/G2/N2). The patient received postoperative radiotherapy and hormonal therapy. Four years later, she presented with an anal polypoid lesion. The mass was removed for biopsy. Immunohistochemical staining suggested a breast origin. Radiotherapy was chosen for this patient, which resulted in complete regression of the lesion. The patient died 3 years after the first manifestation of gastrointestinal metastasis. According to the current literature, we consider the immunohistochemistry features that are essential to support the suspicion of gastrointestinal breast metastasis, and since we consider the gastrointestinal involvement as a sign of systemic disease, the therapy should be less aggressive and systemic. PMID- 19294769 TI - Correlation of p53 over-expression and alteration in p53 gene detected by polymerase chain reaction-single strand conformation polymorphism in adenocarcinoma of gastric cancer patients from India. AB - AIM: To study the alterations in p53 gene among Indian gastric cancer patients and to correlate them with the various clinicopathological parameters. METHODS: A total of 103 gastric cancer patients were included in this study. The p53 alterations were studied by both immunohistochemical method as well as polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) analysis. We only studied four (exon 5, 6, 7, and 8) of the 11 p53 exons. The alterations in p53 were also correlated with respect to various clinicopathological parameters. RESULTS: Among 103 cases, p53 over-expression and alteration were detected in 37 (35.92%) and 19 (18.44%) cases, respectively. Most of the p53 alterations were found at exon 5 (31.54%), followed by exon 6 (26.31%), exon 7 (21.04%) and exon 8 (21.04%). A significant correlation of p53 over-expression was found with p53 alteration (P = 0.000). Concordance between p53 alteration (as detected by SSCP) and over-expression [as detected by immunohistochemistry (IHC)] was found in 75% cases. We found that IHC-positive/SSCP-negative cases accounted for 21% of cases and IHC-negative/SSCP-positive cases accounted for remaining 4% cases. CONCLUSION: Our results show that p53 gene mutations are significantly correlated with p53 protein over-expression, with 75% concordance in over-expression and alteration in the p53 gene, but 25% disconcordance also cautions against the assumption that p53 over-expression is always associated with a gene mutation. There may be other mechanisms responsible for stabilization and accumulation of p53 protein with no evidence of gene mutation that reflect an accumulation of a non-mutated protein, or a false negative SSCP result. PMID- 19294771 TI - Severe dystrophy in DiGeorge syndrome. AB - We present the case history of a 3-year-old girl who was examined because of severe dystrophy. In the background, cow's milk allergy was found, but her body weight was unchanged after eliminating milk from her diet. Other types of malabsorption were excluded. Based on nasal regurgitation and facial dysmorphisms, the possibility of DiGeorge syndrome was suspected and was confirmed by fluorescence in situ hybridization. The authors suggest a new feature associated with DiGeorge syndrome. PMID- 19294772 TI - Resected case of eosinophilic cholangiopathy presenting with secondary sclerosing cholangitis. AB - Eosinophilic cholangiopathy is a rare condition characterized by eosinophilic infiltration of the biliary tract and causes sclerosing cholangitis. We report a patient with secondary sclerosing cholangitis with eosinophilic cholecystitis. A 46-year-old Japanese man was admitted to our hospital with jaundice. Computed tomography revealed dilatation of both the intrahepatic and extrahepatic bile ducts, diffuse thickening of the wall of the extrahepatic bile duct, and thickening of the gallbladder wall. Under the diagnosis of lower bile duct carcinoma, he underwent pylorus-preserving pancreatoduodenectomy and liver biopsy. On histopathological examination, conspicuous fibrosis was seen in the lower bile duct wall. In the gallbladder wall, marked eosinophilic infiltration was seen. Liver biopsy revealed mild portal fibrosis. He was diagnosed as definite eosinophilic cholecystitis with sclerosing cholangitis with unknown etiology. The possible etiology of sclerosing cholangitis was consequent fibrosis from previous eosinophilic infiltration in the bile duct. The clinicopathological findings of our case and a literature review indicated that eosinophilic cholangiopathy could cause a condition mimicking primary sclerosing cholangitis (PSC). Bile duct wall thickening in patients with eosinophilic cholangitis might be due to fibrosis of the bile duct wall. Eosinophilic cholangiopathy might be confused as PSC with eosinophilia. PMID- 19294773 TI - Ileal angiomyolipoma manifested by small intestinal intussusception. AB - Angiomyolipomas (AMLs), a form of benign mesenchymal hamartoma, arise primarily in the kidneys of patients with or without tuberous sclerosis. Extra-renal AMLs are very rare and are most commonly found in the liver. AMLs of the small intestine are exceedingly rare. Here, a case of a 28-year-old man, who presented with ileal intussusception caused by ileal AML is reported. The clinicopathological and immunohistochemical findings of ileal AMLs are discussed and the literature on small intestinal AMLs is reviewed. PMID- 19294774 TI - Congenital absence of the splenic artery and splenic vein accompanied with a duodenal ulcer and deformity. AB - Congenital absence of the splenic artery is a very rare condition. To the best of our knowledge, congenital absence of the splenic artery accompanied with absence of the splenic vein has not been reported. We report a case of the absence of the splenic artery and vein in a 61-year-old woman who presented with postprandial epigastric discomfort. Upper gastrointestinal endoscopy showed a dilated, pulsatile vessel in the fundus and duodenal stenosis. An abdominal computed tomography (CT) scan revealed absence of the splenic vein with a tortuously engorged gastroepiploic vein. Three-dimensional CT demonstrated the tortuously dilated left gastric artery and the left gastroepiploic artery with non visualization of the splenic artery. After administration of a proton pump inhibitor, abdominal symptoms resolved without any recurrence of symptoms during 6 mo of follow-up. PMID- 19294775 TI - Legislation contagion: building resistance. AB - The HIV/AIDS Policy & Law Review recently carried a feature article recounting the spread of problematic new HIV laws in west and central Africa. It outlined less-than-model approaches in the AWARE-HIV/AIDS "model" law and described how its provisions had been replicated in many national HIV laws. At the time of writing that article, eight national HIV laws had been passed in the region. Since that date, the rush to legislate HIV in west and central Africa in ways that do not accord with human rights law or policy has continued unabated. PMID- 19294776 TI - Results of the first North American prescription heroin study are promising. AB - In October 2008, the North American Opiate Medication Initiative (NAOMI) research team released the primary outcomes of a randomized controlled trial aimed at testing whether the provision of pharmaceutical-grade heroin under medical supervision benefits people suffering from chronic opiate addictions who have not benefitted from other treatments. The treatment phase was completed in June 2008. Retention and response rates were high, suggesting that heroin-assisted therapy is a safe and highly effective treatment for people with chronic heroin addiction. PMID- 19294777 TI - New excessive demand policy for immigration applicants. AB - Under a new policy adopted in September 2008, Citizenship and Immigration (CIC) officers must now consider all evidence presented by an immigration applicant before making a decision of inadmissibility due to excessive demand on social services. Evidence regarding both ability and intent to mitigate the cost of social services in Canada must be considered, if presented. PMID- 19294778 TI - British Columbia adopts E-Health Act. AB - On 29 May, 2008, Bill 24 passed into law in British Columbia. The E-Health (Personal Health Information Access and Protection of Privacy) Act puts in place a framework for the establishment of government databases of personal health information. PMID- 19294779 TI - (Most) political parties respond to questionnaire on AIDS issues during federal election. AB - During the recent federal election campaign, the Canadian HIV/AIDS Legal Network and the Interagency Coalition on AIDS and Development sent out a questionnaire to the leaders of the five main federal political parties asking about their parties' positions on several key issues related to HIV/AIDS. The Liberals, New Democratic Party (NDP), Bloc Quebecois and Green party responded. The Conservative Party did not. PMID- 19294780 TI - Immigration and Refugee Protection Act amended. AB - With the passage of Bill C-50 (Budget Implementation Act, 2008) in June 2008, proposed changes to the Immigration and Refugee Protection Act (IRPA) contained in Part 6 of Bill C-50, were adopted. PMID- 19294781 TI - Federal Government announces new strategy to eliminate illegal drugs in federal prisons. PMID- 19294782 TI - Mexico: general recommendations issued against mandatory HIV testing and discharge from armed forces. AB - In September and October 2008, Mexico's National Human Rights Commission (NHRC) issued three separate Recommendations addressing the practice of mandatory HIV testing and discharging HIV-positive personnel from Mexico's armed forces. PMID- 19294783 TI - Cambodia: human trafficking legislation threatens HIV response. AB - In February 2008, Cambodia's new Law on the Suppression of Human Trafficking and Sexual Exploitation was promulgated and went into effect. The law criminalizes sex for money, public soliciting for prostitution and many forms of financial transactions connected to sex work. The law has been criticized for conflating sex work and trafficking. PMID- 19294784 TI - U.S.: interim step towards elimination of HIV travel ban. AB - On September 29, 2008, the U.S. Department of Homeland Security (DHS) promulgated new regulations streamlining the travel authorization process for people living with HIV wishing to enter the United States as non-immigrants. PMID- 19294785 TI - Global: review of injecting drug use and HIV prevalence among users. AB - In September 2008, The Lancet published one of the few worldwide estimates of the number of people who inject illicit drugs and of the prevalence of HIV among this population. PMID- 19294786 TI - Uganda: civil society expresses concern about HIV bill. PMID- 19294787 TI - China: Hu Jia wins human rights prize. PMID- 19294788 TI - Vancouver's supervised injection facility granted constitutional exemption from federal drug law. AB - In a landmark decision on 27 May 2008, the B.C. Supreme Court held that because Insite--North America's only supervised injection facility (SIF)--provided health care, denial of access to Insite and safe injection would threaten individuals' rights to life, liberty and security of the person. PMID- 19294789 TI - HIV-positive man from DRC deemed a "person in need of protection". AB - In an in camera hearing on 9 July 2008, the Refugee Protection Division of the Immigration and Refugee Board of Canada, accepted that the manner in which the government of the Democratic Republic of the Congo (DRC) dealt with the HIV epidemic in its country posed a "risk to life" to an HIV-positive man from that country who was applying for asylum in Canada. PMID- 19294790 TI - Federal Court sets aside decision denying Mexican couple's claim of persecution. AB - On 11 April 2008, the Federal Court allowed the application of Castillo Ramirez and Viccon Palacios for judicial review of a decision by the Refugee Protection Division (RPD) of the Immigration and Refugee Board, which found that the applicants were neither Convention refugees nor persons in need of protection. PMID- 19294792 TI - Criminal law and cases of HIV transmission or exposure. Developments in Johnson Aziga case. PMID- 19294791 TI - Court dismisses HIV-positive man's application for review of decision not to defer removal from Canada. AB - On 7 July 2008, Wilson Gumbura sought judicial review of a decision by an enforcement officer not to defer his removal from Canada until a decision had been made on his humanitarian and compassionate application. PMID- 19294793 TI - Manitoba court considers condom use and HIV viral load in analysis of "significant risk". PMID- 19294794 TI - New trial ordered in B.C. case of non-disclosure of HIV-positive status. PMID- 19294795 TI - Man acquitted of sexual assault for non-disclosure of HIV status before oral sex. PMID- 19294796 TI - The gender of resistance in Communist Poland. PMID- 19294797 TI - The historical evolution of the cutting-in pattern, 1798-1967. PMID- 19294798 TI - The mystery of the disappearing pursers' typists. PMID- 19294799 TI - The responses of American Indian children and Irish children to the school, 1850s - 1920s. PMID- 19294800 TI - The sensibility of comfort. PMID- 19294801 TI - The Siegen affair. PMID- 19294802 TI - Practical Neurology. PMID- 19294803 TI - Trust, pawnship, and Atlantic history: the institutional foundations of the old Calabar slave trade. PMID- 19294804 TI - Clinical questions need perceptive answers. PMID- 19294805 TI - [America incorporated: cannibalism as utopia in imagery of early America]. PMID- 19294806 TI - [Clergy and the middle class in Limousin during the first decades of the 19th century]. PMID- 19294807 TI - [Marseilles elites and worker housing at the end of the 19th century: the Bouches du-Rhone Savings Bank's working-class housing in the Capelette neighborhood]. PMID- 19294808 TI - [Falaise outside the railroad network, 1846-67]. PMID- 19294809 TI - [Urban itineraries: changes of residence within Milan in the 19th century]. PMID- 19294810 TI - Understanding the metabolic responses to sepsis. PMID- 19294811 TI - Liver function tests. PMID- 19294812 TI - How to deal with torture victims. PMID- 19294813 TI - Understanding clinical variability in response to asthma treatment. PMID- 19294814 TI - Solitary confinement. An introduction to the Istanbul Statement on the Use and Effects of Solitary Confinement. PMID- 19294815 TI - The Istanbul Statement on the Use and Effects of Solitary Confinement. PMID- 19294816 TI - Clinical audit 1: a guide for the foundation year doctor. PMID- 19294817 TI - Supervision: what the foundation year doctor needs to know. PMID- 19294818 TI - Japes and jackanapes: on being a junior hospital doctor in the 1970s. PMID- 19294819 TI - Being a junior doctor today. PMID- 19294820 TI - So you want to be ... an ophthalmologist. PMID- 19294821 TI - Protection planning: be prepared. PMID- 19294822 TI - Interview with outgoing President of AROI, by Meena Tiwari . PMID- 19294823 TI - A critical analysis of the benefits and limitations of an applied degree in undergraduate nursing education. PMID- 19294824 TI - Improving psychological and psychiatric aspects of palliative care: the national consensus project and the national quality forum preferred practices for palliative and hospice care. AB - As patients with terminal disease enter into the final stage of their illness, psychiatric symptoms and psychological responses to the disease contribute to overall suffering of both patient and family. Until recently, no nationally accepted guidelines or practices had been established to support assessment and management of this type of suffering. In 2007, the National Quality Forum published A National Framework and Preferred Practices for Palliative and Hospice Care Quality that included a list of preferred practices for assessing and treating symptoms of psychiatric illness, anticipatory grief and psychologic distress prior to death, and bereavement after the death, of the patient. While specialized care may be provided to patient and families in the context of advanced disease, all clinicians involved in palliative and end-of-life care are responsible for having a basic understanding of effectively managing psychologic and psychiatric aspects of this care. Evidence from current literature supports these best practices. PMID- 19294825 TI - Psychological adjustment to conjugal bereavement: do social networks aid coping following spousal death? AB - This research sought to investigate the role of social networks in coping and adjustment to spousal bereavement. A total of 198 Xhosa-speaking participants, drawn from the rural areas of the Eastern Cape Province, South Africa, took part in the study. Quantitative data analyses revealed significant correlations between perceived social support on Social Support Appraisals scale and the anxiety scores on the Beck Depression Inventory-Second Edition and the problem solving coping strategy scale on the Coping Strategy Indicator. Regression analyses revealed that depression contributed to increased social support appraisals, while anxiety was a significant negative predictor of the participants' experience of perceived social support. The authors highlight the need for bereavement intervention programs in the rural areas to be geared toward psycho-educating the conjugally bereaved individuals, within the socio-cultural framework, to continuously evaluate their support structures and strengthen ties with social networks that render support, instrumental assistance, and advice in times of need. PMID- 19294826 TI - Death notification for families of homicide victims: healing dimensions of a complex process. AB - Family bereavement by homicide is one of the most challenging facets of crime victim intervention a medical care provider, mental health clinician, or law enforcement officer will face. But the process of healing can begin at the outset through intelligent, sensitive, competent, and compassionate death notification and body identification by trained and dedicated professionals. A practical system for death notification and body identification is presented that can be adapted and modified to the needs of law enforcement, military mental health, and notifiers from related disciplines; it can also be adapted to death notification in sudden, accidental deaths. Throughout, the emphasis is on how notifiers may often be the de facto first responders in the struggle to help traumatically bereaved family members survive their ordeal and reclaim their lives. PMID- 19294827 TI - Caregivers' communication with patients about illness and death: initial validation of a scale. AB - Caregivers of terminal cancer patients experience substantial communication difficulties with their loved ones about their illness. Existing communication scales focus on communication as perceived by the patient and do not include items that refer to his close death. The current study describes the development and initial validation of an instrument aimed at measuring caregivers' communication with patients about their illness and death. Two hundred thirty-six bereaved primary caregivers of cancer patients were recruited over a period of 18 months. The psychometric properties of the scale were explored by confirmatory factor analysis. Results provided support for a one factor solution. The discriminant and convergent validity of the responses to the scale were also supported. Future studies should further establish the validity of the scale with larger sample sizes and different populations of caregivers of patients with other terminal diseases. PMID- 19294828 TI - Stuff happens! PMID- 19294829 TI - Sodium hypochlorite accident--a complication of poor access cavity design. AB - A case is presented of soft tissue damage resulting from inadvertent injection of sodium hypochlorite through a perforation created during root canal treatment of a mandibular incisor. The importance of access cavity design to help avoid such complications is highlighted and the anatomy of the mandibular incisor is discussed. Measures to minimize the risk of a hypochlorite accident and its potentially serious complications are proposed. PMID- 19294830 TI - Inadvertent injection of sodium hypochlorite into the periradicular tissues during root canal treatment. AB - This paper describes a sodium hypochlorite (NaOCl) incident occurring during routine endodontic treatment of a patient who presented with chronic periapical periodontitis of a maxillary canine tooth and discusses the immediate and late treatment of this case. PMID- 19294831 TI - Fracture of a dental needle during administration of an inferior alveolar nerve block. AB - We report a case of a 42-year-old female who was referred to our department for the management of a fractured dental needle following administration of an inferior alveolar nerve block. The fractured needle was successfully removed under general anaesthesia. We also suggest some guidelines for preventing this unfortunate event. PMID- 19294832 TI - Oro-facial thermal injury caused by food heated in a microwave oven. AB - Burns to the oral mucosa usually result from the accidental ingestion of hot food or beverages. The burns are usually of short duration and little consequence. The widespread use of microwave ovens, however, has added a new dimension to the problem. Microwave ovens heat food much quicker than a conventional oven, but they produce uneven heating within the food and extremely high temperatures can be reached. We describe two cases of patients who suffered inadvertent injury to the oral mucosa from the ingestion of microwave-heated food. PMID- 19294833 TI - The incidental finding of a retained gunshot pellet. AB - An 80-year-old woman presented to the restorative department at the Charles Clifford Dental Hospital in Sheffield for a routine review of her dental health with no reported trauma, pain or complications relating to the oral environment. A routine periapical radiograph showed the presence of a foreign object in the right maxillary second premolar region. An unusual case is reported of a retained gunshot pellet lying in the upper buccal mucosa for 70 years without the patient experiencing any pathological signs or symptoms. PMID- 19294834 TI - Paracetamol overdose in a teenager following placement of a fixed appliance. AB - Pain or discomfort is common following placement or adjustment of orthodontic appliances. This case report describes an accidental paracetamol overdose in a 13 year-old girl following the initial placement of fixed appliances. PMID- 19294835 TI - A review of aggressive periodontitis and an associated case report. AB - Aggressive periodontitis is a debilitating oral disease that results in rapid destruction of the periodontal tissues. It has recently been reclassified and ongoing efforts are now being made to understand its pathogenesis and record its prevalence in the population. This case report gives a brief outline of studies that have investigated why this disease tends to occur in otherwise young healthy adults. The mechanisms behind such extensive periodontal damage are discussed. A case report of a patient with aggressive periodontitis, treated in the Dublin Dental School and Hospital, is presented and the rationale behind his treatment is discussed. PMID- 19294836 TI - Common things are common: a case series of oral foreign bodies in paediatric patients. AB - Reports of foreign bodies in the oral cavity are few in number. Three cases of children of varying ages, presenting with oral foreign bodies, and their subsequent diagnosis and management, are described. The importance of considering foreign bodies, as part of a differential diagnosis in paediatric patients, where aetiology is uncertain and clinical appearance is unusual, is highlighted. PMID- 19294837 TI - Physical signs for the general dental practitioner. Case 61. Erythema multiforme. PMID- 19294838 TI - Aspects of human disease. 29. Rheumatoid arthritis. PMID- 19294839 TI - [Resources of Chinese Herbs Sang (Morus sp.) in Sichuan and strategies for development and utilization]. AB - A comprehensive analysis was conducted on resources composition of Chinese Herbs Sang (Morus sp.) in Sichuan using survey data and related literature. The original plants, germplasm collections, cultivation areas, main cultivated varieties and production sale of crude drugs of Sang in Sichuan were clearly expounded. Strategies for development and utilization of Sichuan mulberry resources were suggested. PMID- 19294840 TI - [Development and evaluation of terpene penetration enhancers]. AB - Major advantages of transdermal drug delivery system (TDDS) include avoiding of drug degradation in hepatic and gastrointestinal, predetermining release rate and blood drug level, reducing administration frequency and adverse reactions, and increasing patient compliance. But its application was limited by stratum corneum barrier and low skin permeability of drugs. Terpene penetration enhancers are low toxic and can improve the skin permeability and enhance the permeating veloc both of hydrophilic and lipophilic drugs. Terpenes can also significantly enhance the cumulative release amount of drug at its low concentration comparing with other synthetic penetration enhancers, and play an important role in the TDDS. This review presented the source, classification, mechanism and applications of terpenes, combined use with other enhancers and methods. Optimization, evaluation and prospective applications of terpene penetration enhancers were also discussed. PMID- 19294841 TI - [Progress in studies of mechanism of anti-fibrosis traditional Chinese drugs and effective components]. AB - The progress in the research of pharmacological mechanism of anti-fibrosis traditional Chinese drugs and the effective components is summarized. It's demonstrated by pharmacological experiments that anti-fibrosis traditional Chinese drugs can inhibit the cell proliferation, regulate the cytokines and ECM, and intervente in the signal transduction. PMID- 19294842 TI - [Primary studies on biological characteristics of Dorysthenes hydropicus]. AB - OBJECTIVE: To study the biological characteristics of Dorysthenes hydropicus in the farm of Cirtus grandis, and offer scientific evidence for prevention and controlling of D. hydropicus. METHOD: Indoor-rearing and light trap were applied to study the biological characteristics, development course and harmful effect of D. hydropicus. RESULT: D. hydropicus reproduces one generation in 1-2 year in Guangdong province, and overwinters in the form of larvae. Its imago comes out of the earth mainly in late May after mature. The body length has great individual diversity normally ranged from 25-60 cm, It also shows strong phototaxy. One lamp can trap more than 2 000 of them per night. Female imago has a large egg load with the maximum amount of 543. The eggs hatching is in depth of 1-3 cm soil. The dominant hatching period of egg is from late June to early July, and hatchability is over 85%. The living space of larva ranges from 15-60 cm in soil. D. hydropicus has caused serious harm and lead to thousands of Cirtus grandis trees death every year. CONCLUSION: Dorysthenes hydropicus showed serious threat to the growth of Cirtus grandis and should be prevented and controlled. PMID- 19294843 TI - [Comparative study on botanical morphologic character of different cultivated types medicinal Chrysanthemum morifolium]. AB - OBJECTIVE: Botanical character of different cultivated types medicinal Chrysanthemum morifolium were observed and compared, which can offer reference for its genetic diversity research and breeding. METHOD: Based on the random blocks field experiments design, the main character of leaf and capitulum of medicinal C. morifolium were observed, it was studied on that the botanical character of different cultivated types medicinal C. mortfolium in China were comparatively study with statistical means. RESULT AND CONCLUSION: There are some different of botanical character among different cultivated types medicinal C. morifolium in China, which can be divided into 7 colonies according as its diversity of the botanical and morphological character. The sawtooth number of lamina, leafstalk length and leaf division number are the important laminae morphologic character of different types medicinal C. morifolium, which are relative independent characters. PMID- 19294844 TI - [Effects of shading on photosynthetic characteristics of Pinellia ternata leaves]. AB - OBJECTIVE: To study the effects of shading on photosynthetic physiology and chlorophyll fluorescence of Pinellia ternata. METHOD: Plant growth, chlorophyll content, net photosynthetic rate (P(n)) and chlorophyll fluorescence in P. ternata were investigated under different shading treatments (0%, 70% and 90%) when it grew about 15 cm high. RESULT: The results showed that fresh weight of a tuber, height, leaf length, width, leaf area, specific leaf area (SLA) and contents of chlorophyll content were enhanced after shaded, and chlorophyll a/b rate declined. Compared with control, net photosynthetic rate, light compensation point (LCP) and light saturation point (LSP) of P. ternata decreased after shading, but apparent quantum yield (AQY) increased; quantum yield of PS II (PhiPS II), minimal fluorescence (F(o)), maximal fluorescence (F(m)), intrinsic photochemical efficiency of PS II (F(v)/F(m)) and photochemical quenching coefficient (qP) were enhanced. CONCLUSION: Compared with control, all data indicated that there were distinctive differences between the height, SLA, chlorophyll content, P(n) and chlorophyll fluorescence characteristics under the shading treatments (70% and 90%), the fresh weight of a tuber increased after 70% shading, and provided better environmental conditions for the growth of P. ternata. PMID- 19294845 TI - [Effects of S-3307 on the yield and main ingredients of Alisma plantago aquatica]. AB - To study the effect of S-3307 on the yield and main ingredients of Alisma plantago-aquatica. METHOD: The contents of 24-acetyl alisol A and the 23-acetyl alisol B in tuber were determined by HPLC. RESULTS: The contents of 24-acetyl alisol A and the 23-acetyl alisol B as well as yield were significantly increased in all groups applied with different concentrations of S-3307 comparing with control group. The optimal concentration of S-3307 was 80 mg x kg(-1). The residues of S-3307 was detected under 0.316 8 mg x kg(-1) (detecting limit). CONCLUSION: The optimal concentration of S-3307 is 80 mg x kg(-1), it reached the best result when applied 36 d after seedling. PMID- 19294846 TI - [Variation of ginsenosides in infected roots of American ginseng (Panax quinquefolium)]. AB - To study the infected root of Panax quinquefolium on the contents of ginsenosides. METHOD: The contents of three major ginsenosides Rg1, Re and Rb1 were determined by HPLC compared quantitatively between the different degree infected roots and normal root in the phloem and xylem. RESULT: Rg1 in phloem and xylem of varying degrees infected root showed no significant difference, but Rb1 decreased 26.3% and 28.3% respectively in medium and serious infected roots comparing to normal root. Re in phloem of seriously infected roots decreased in xylem significantly. CONCLUSION: The results indicate that the variation of ginsenosides in different degrees infected roots exists and the proportion of Rg1, Re and Rb1 in the total ginsenosides changes. PMID- 19294847 TI - [Comparison of two kinds of methods to prepare Chinese medicine Danggui Buxue formula pellets]. AB - The pellet-preparating theories and processes of two kinds of methods of extrusion-spheronization and fluid-bed coating method were compared. The micromeritic properties, yield and in vitro dissolution of Danggui Buxue pellets prepared by the two methods were determined. Results showed that the Danggui Buxue pellets prepared by extrusion-spheronization were all spheral with smooth surface, the yield of those pellets was higher and in vitro dissolution of those pellets was more satisfactory. So the two kinds of methods were all suitable to prepare Dang-gui-bu-xue pellets, but Extrusion-spheronization was superior to fluid-bed coating method. PMID- 19294848 TI - [Chemical constituents from flowers of Carthamus tinctorius]. AB - OBJECTIVE: To study the chemical constituents from the flowers of Carthamus tinctorius. METHOD: Compounds were isolated by chromatographic techniques. Their structures were elucidated by spectral methods. RESULT: Ten compounds were identified as 7,8-dimethylpyrazino [2,3-g] quinazolin-2, 4-(1H, 3H) -dione (1), adenosine (2), adenine (3), uridine (4), thymine (5), uracil (6), roseoside (7), 4'-O-dihydrophaseic acid-beta-D-glucopyranoside methylester (8), 4-O-beta-D glucopyranosyloxy-benzoic acid (9) and p-hydroxybenzoic acid (10). CONCLUSION: Compounds 1 and 8 were isolated from natural plants for the first time, and compounds 7, 9 and 10 were isolated from this plant for the first time. PMID- 19294849 TI - [Chemical constituents from dried sorophore of cultured Cordyceps militaris]. AB - OBJECTIVE: To investigate the chemical constituents of the dried sorophore of cultured Cordyceps militaris. METHOD: Compounds were isolated and purified by macroporous adsorption resin and silica gel column chromatography. Their chemical structures were elucidated on the basis of physicochemical properties and spectral data (IR, FAB-MS, 1H-NMR and 13C-NMR). RESULT: Nine compounds were isolated and identified as: ergosta-4, 6, 8 (14)-tetraen-3-one (1), citrostadienol (2), tetracosanoic acid 2, 3-dihydroxypropyl ester (3), ergosterol (4), ergosterol peroxide (5), ergosta-7, 22-dien-3beta, 5alpha, 6beta-triol (6), cordycepin (7), adenosine (8), N-(2-hydroxyethyl) adenosine (9), respectively. CONCLUSION: Compounds 1-3, 6, 9 were separated from the sorophore of cultured C. militaris for the first time. PMID- 19294850 TI - [Studies on chemical constituents in roots and rhizomes of Notopterygium incisum]. AB - OBJECTIVE: To study the chemical constituents in the roots and rhizomes of Notopterygium incisum. METHOD: The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by NMR and MS evidences. RESULT: Twenty-four compounds were obtained and identified as isocalamenediol (1), pregnenolone (2), beta-sitosterol (3), bergapten (4), bergaptol (5), cnidilin (6), isoimperatorin (7), bergamottin (8), demethylfuropinnarin (9), nodakenetin (10), 7'-O-methylnotoptol (11), notopterol (12), notoptol (13), pabulenol (14), phenethylferulate (15), p-hydroxyphenethyl anisate (16), trans-ferulic acid (17), nodakenin (18), sucrose (19), 2-methoxy-4 (3-methoxy-1-propenyl)-phenol (20), 3, 4, 5 trimethoxy-trans-cinnamic acid (21), p-methoxycinnamic acid (22), 4-acetoxy-3-methoxy-trans-cinnamic acid (23), and p hydroxy-trans-cinnamic acid (24). CONCLUSION: Compounds 1, 14, 20-24 were isolated from the roots and rhizomes of N. incisum for the first time. Compounds 1 and 14 are sesquiterpenoid and coumarin, respectively, and 20-24 were phenylpropanoid compounds. PMID- 19294851 TI - [Phenolic components from herbs of Dendrobium aphyllum]. AB - OBJECTIVE: To study the phenolic constituents of Dendrobium aphyllum. METHOD: Various chromatographic techniques were used to isolate and purify the constituents, their physico-chemical properties and spectral data were employed to elucidate their structures. RESULT: Nine bibenzyls and two benzylethanyl compounds were isolated and identified as: moscatilin (1), gigantol (2), batatasin (3), tristin (4), 3, 5, 4'-trihydroxylbibenzyl (5), 3, 5-dimethoxyl-4, 4'-dihydroxylbibenzyl (6), moscatin (7), 2, 4, 7-trihydroxyl-9, 10 dihydrophenanthrene (8), hircinol (9), 2-(4-hydroxyphenyl) ethyl-beta-D glucopyranoside, salidroside (10) and p-hydroxylbenzylacetic acid (11). CONCLUSION: All compounds were obtained firstly from the plant, and the compounds 10 and 11 were isolated in this genus for the first time. PMID- 19294852 TI - [Studies on isolation and identification of flavonoids in herbs of Agrimonia pilosa]. AB - OBJECTIVE: To study the chemical constituents in Agrimonia pilosa. METHOD: The compounds were isolated and purified by various column chromatographic methods and elucidated on the basis of chemical and spectroscopic evidences. RESULT: Nine flavonoids were obtained and identified as tiliroside (1), kaempferol 3-O-alpha-L rhampyranoside (2), quercetin 3-O-alpha-L-rhampyranoside (3), quercetin 3-O-beta D-glucopyranoside (4), kaempferol 3-O-beta-D-glucopyranoside (5), kaempferol (6), apigenin (7), luteolin (8), quercetin (9). CONCLUSION: Compounds 1-3, 5, 6 and 8 were isolated from this plant for the first time. PMID- 19294853 TI - [Study on LC-MS fingerprint for quality assessment of Aristolochia manshuriensis with chemical pattern recognition]. AB - OBJECTIVE: To analyze LC-MS fingerprints of Aristolochia manshuriensis for quality assessment with two different chemical pattern recognition models. METHOD: LC-MS fingerprints of A. manshuriensis were established from 24 batches of samples from different habitats. SIMCA and Clustering analysis were used to compare the parameters of the 29 common peaks. RESULT: Two methods had good consistency, while they reflected the inherent sample information from different perspectives, respectively. CONCLUSION: Modern equipment analysis technology and multivariable chemical pattern recognition would be an efficient way for quality control and variety identification of A. manshuriensis. PMID- 19294854 TI - [Study on fingerprint of Cortex Fraxini with HPLC]. AB - OBJECTIVE: To establish fingerprint of Cortex Fraxini and provide reference for quality evaluation of Cortex Fraxini. METHOD: Chromatographic experiments were performed on Agilent Extend C18 column (4.6 mm x 250 mm, 5 microm), eluted with methanol and water, containing 0.4% acetic acid as the mobile phases in gradient elution. The detection wavelength was 0-60 min, 340 nm; 6-74 min, 254 nm; 74-75 min, 340 nm, and the flow rate was 1.0 mL x min(-1). Forty samples in four varieties of Cortex Fraxini were detected to establish fingerprints, respectively. RESULT: Every parameter of the method validation complied with related rules and regulations. There were 15 common peaks in the fingerprint of 10 Fraxinus rhynchophylla samples, eleven common peaks in the fingerprint of 10 F. chinensis var. acuminata. samples, and in the fingerprint of 10 F. chinensis samples. Nineteen common peaks in the fingerprint of 10 F. stylosa samples. There were 5 common peaks in the fingerprints of 40 Cortex Fraxini samples. The similarity factors of the 10 samples of every species were all more than 0.96 compared with the control fingerprint. The similarity of the 40 Cortex Fraxini samples was more than 0.90. Four effective constituents and one unknown constituent were found in 40 samples. CONCLUSION: The fingerprints of F. rhynchophylla bark, F. chinensis bark, F. chinensis var. acuminata bark, F. stylosa bark and Cortex Fraxini were established. The methodological evaluation showed that the results were in accord with the technology requirements of chromatography fingerprint, and it laid a good foundation for quality control of Cortex Fraxini. PMID- 19294855 TI - [Determination of zizybeoside II of Ziziphus jujuba by HPLC]. AB - OBJECTIVE: To develop a quantitative method for determination of zizybeoside II in Ziziphus jujuba. METHOD: The samples were separated at 30 degrees C on a Zorbax SB-C18 column eluted with methanol-water (20 : 80) as the mobile phase. Flow rate was set at 1.0 mL x min(-1) and the detection wavelength was set at 210 nm. RESULT: The calibration curve was linear within the range from 0.046 to 0.582 microg (r = 0.999 9) and the average recovery was 97.2%. 12 batches of the crude drugs purchased from different areas were determined and the contents of zizybeoside II in Fructus Jujubae were fluctuated from 0.013% to 0.041%. CONCLUSION: The method is simple, repeatable and could be used for the quality control of Z. jujuba. PMID- 19294856 TI - [Regulation function of Qingnao drop pilula to MARCKS mRNA express changes in acute cerebral ischemia hippocampus]. AB - OBJECTIVE: To observe the contribution of Qingnao drop pilula to the alteration of myristoylated alanine-rich C kinase substrate (MARCKS) mRNA expression in acute multi-infarction hippocampus. METHOD: Rat models of acute multi-infarction were established by injecting the embolus of blood powder through the right external carotid arteryinto the internal carotid artery, rats were randomly divided into five groups (n = 12 in each): normal, sham operation, model, Chinese medicine treatment, and Western medicine treatment. Qingnao drop pilula (133.28 mg x kg(-1)), nimodipine (7.25 mg x kg(-1)) were administered respectively to Chinese medicine treatment group and Western medicine treatment group by gavage, equal volume of normal saline were given to three groups. Rats were treated with drugs starting at 3rd day before the operation, one time per day. Observing morphologic changes in hippocampus by optical microscope and electron microscope. Detecting expression level of MARCKS mRNA in hippocampus by semi-quantification PCR method. RESULT: Hippocampus cells arrange tidy, administrative levels were compactness in normal group, which cells differentially impaired in model group, Chinese medicine treatment group and Western medicine treatment group. Hippocampus cells damage of Chinese medicine treatment group have more reckless than the model group in histopathology. The MARCKS mRNA were expressioned in model group vs medication treatment groups, in Chinese medicine treatment group vs the model group. CONCLUSION: Qingnao drop pilula can alleciate histomorphology lesion of hippocampus when occurring acute multi-infarction, to turn slower MARCKS mRNA expression, may play a neuroprotective effect role through accommodating PKC-MARCKS signal transduction system. PMID- 19294857 TI - [Effect of isopropyl 3-(3,4-dihydroxyphenyl) -2-hydroxypropanoate on rat pulmonary artery smooth muscle]. AB - OBJECTIVE: To investigate the effect of isopropyl 3-(3,4-dihydroxyphenyl)-2- hydroxypropanoate on vascular smooth muscle. METHOD: Isolated rat pulmonary artery was perfused and the tension of the vessel was measured, the effect of isopropyl 3-(3, 4-dihydroxyphenyl)-2-hydroxypropanoate on the pulmonary artery precontracted by noradrenaline (NE) and concentration-response curves of 5 hydroxytryptamine (5-HT), endothelin-1 (ET-1), U46619 and KCl was also observed. RESULT: Isopropyl 3-(3,4-dihydroxyphenyl) -2-hydroxypropanoate exerted relaxation effect on the endothelium-intact artery precontracted by NE in a concentration dependent manner, which was inhibited with denuded endothelium. The right-shift of the concentration-response curves of 5-HT, ET-1, U46619 and KCl. CONCLUSION: Isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate have relaxation action on rat pulmonary artery in the way of endothelium-dependance, the mechanism of relaxation action by isopropyl 3-(3,4-dihydroxyphenyl) -2-hydroxypropanoate may be related to calcium channels. PMID- 19294858 TI - [Study on drug-guide effect of Achyranthes bidentata in Sanmiao pill in arthritic rats]. AB - OBJECTIVE: To study the drug-guiding mechanism of Achyranthes bidentata from Sanmiao pill in arthritic rats. METHOD: The rats were treated by Ferud's complet adjuvant and hot water -bath to establish rat adjuvant arthritis model, then the model rats were divided into three groups, model group, Sanmiao pill groups with A. bidentata. (18 g x kg(-1)) and without A. bidentata. (10 g x kg(-1)), respectively. The heart and foot joints were washed and homogenated to determine the berberine concentration by HPCL in different time after ig and the foot edema was tested by volume method. The pathological changes were observed and hemorheologic parameters were also tested. RESULT: The berberine concentration of foot joint was significantly higher in 2, 4, 6 hour and 14 day in the rats with A. bidentata. The berberine concentration ratio of foot joint and heart was significantly higher in rats with A. bidentata. pharmacodynamic researches showed that A. bidentata could enhance the edema inhibition effect of Sanmiao pill. Hemorheologic researches showed that A. bidentata. could significantly improve the blood viscosity of model rats, the blood high shear viscosity, the blood low shear viscosity and the plasma viscosity were (6.47 +/- 0.57), (9.28 +/- 1.2), (1.94 +/- 0.19) mPa x s respectively. CONCLUSION: A. bidentata. could facilitate the targeted tissue distribution of berberine. The effect was correlative with its blood viscosity improvement. PMID- 19294859 TI - [Study on effect of promoting blood circulation drugs components in treating unstable angina in patients with blood stasis inflammatory levels]. AB - OBJECTIVE: To discussion the effects of Huoxue components of effective drug in treating unstable angina in patients with blood stasis WBC (WBC), C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha). METHOD: one hundred and twenty cases of unstable angina were randomly divided into the conventional therapy group, component compatibility group, Pieces group and Xuesaitong group 4 groups, each with 30 cases. Observation of patients before and after treatment of clinical efficacy, blood lipid indicators and the indicators changes. RESULT: Component compatibility group after treatment clinical marked improvement in conditions, and the WBC, CRP, IL-6 and TNF-alpha, TC, TG levels lower than before treatment, there were significant differences (P < 0.05), and lower than the other three groups After treatment (P < 0.05). And HDL-C after treatment than before treatment increased, there were significant differences (P < 0. 05). CONCLUSION: Huoxue-effective component compatibility can be effective treatment of unstable angina blood stasis, and could inhibit the inflammatory level. PMID- 19294860 TI - [Study of molecular mechanism of tanshinone II A inducing differentiation in acute promyelocytic leukemia NB4 cells]. AB - OBJECTIVE: To investigate molecular mechanism of tanshinone II A inducing differentiation and apoptosis in acute promyelocytic leukemia NB4 cells. METHOD: NB4 cells were cultured in vitro and treated with tanshinone II A and observed cellular morphology, cell category and the cellular proliferation. DNA microarray technique was used to analyze the gene expression profiles of NB4 cells induced by tanshinone II A. RESULT: 92.8% of NB4 cells treated with 0.5 mg x L(-1) tanshinone II A were induced into mature neutrophils, in which myetocytes and melamyetocytes were 27.0%, banded and segmented neutrophits 68.2%. Cell growth were inhibited. cDNA microarray showed the enormously expressed 183 genes including 23 differentiation associated genes, and other interrelated genes. CONCLUSION: Tanshinone II A inducing differentiation in NB4 cells may be via regulation of many kinds of genes, especially differentiation associated genes expression. This partially explained the molecular mechanism of tanshinone II A inducing differentiation. PMID- 19294861 TI - [Evaluation of immune regulation effects of Chinese medicinal formulae based on immunomics of traditional Chinese medicine]. AB - OBJECTIVE: To investigate the possibility of evaluation the Chinese medicinal formulae with immune regulation effects according to the immunomics of traditional Chinese medicine. METHOD: The possibility of Chinese medicinal formulae with immune regulation effects was obtained from the discriminant equation of the Chinese medicine with immune regulation effects. The original articles about the Chinese medicinal formulae with immune regulation effects were found since 1980 in CNKI. Using literatures determinate immune effects. RESULT: There were 53 kinds, that with calculated determination, from total 60 Chinese medicinal formulae with literatures determinate on immune regulation effects. The coincidence was 88% (53/60). The discriminant score was 0. 89 +/- 0. 03 in group one (there were two or more than two articles have been published to support the medicines immune effects and the articles were finished by different unites independently) and was 0.60 +/- 0.29 in group of random Chinese medicinal formulae. There was a significant deviation (P < 0.05). CONCLUSION: It was proved preliminarily that the principle of calculation presented in this paper is valuable for the immunomics of traditional Chinese medicine. PMID- 19294862 TI - [Experimental [corrected] study of hypoglycemic activity of conduritol A of stems of Gymnema sylvestre]. AB - OBJECTIVE: To investigates the mechanism of hypooglycemic effect of conduritol A of stems of Gymnema sylvestre. METHOD: Fourteen days later after administration, observation is taken on the change of these mice and rats weight, the FBG, TG, CHO, SOD, MDA, INS, TNF in serum were also detected with enzymology method and Radioimmuoassay method. Take the liver to determine the disposal of glucose. Take the pancreas to do the HE and immunohistochemistrial staining, and show pancreas islet beta-cell. Calulate thymus, pancreas, splenica index. RESULT: Compared with diabetic model mice, high and middosage of conduritol A could remarkably reduce fasted blood sugar in diabetic rats induced by alloxan (P < 0.01). Significantly increase the level of serum insulin (P < 0.05). Activity of SOD was obviously increased, and amount of MDA was obviously decreased (P < 0.05). The amount of conduritol A disposal of glucose was obviously increased (P < 0.05). Significantly increase thymus, pancreas, splencia index (P < 0.01 or 0.05); inhibited the atrophy of thymus, pancreas, splencias of the diabetic rats induced by alloxan. Compared with diabetic model group, cell structure and form of conduritol A had been some way improved. The immunohistochemistry results showed that beta-cells numbers of pancreas in each conduritol A group were more than those in the model group. CONCLUSION: Conduritol A could have an effect on regulating the metabolism of blood lipid, free-radical scavenging, enhancing the antioxidant ability, potentiating immune function. Promoting synthesis of hepatic to decrease fasted blood suger. PMID- 19294863 TI - [Effect of Zhizhuwan on gastrointestinal peptide concentrations in plasma of diabetic gastroenteropathy with constipation patients]. AB - OBJECTIVE: To determine the therapeutic efficacy and elucidate the probable pharmacological mechanism of Zhizhuwan (ZZ), a known formula for GI/Dysmotility and constipation in Traditional Chinese medicine, in managing constipating diabetes mellitus (DM). METHOD: Subjects, composed of both males and females, ranged from 55 to 76 years of age, and had constipating DM. Subjects with severe colon, rectum, heart, renal, or hepatic disease, obesity and those on medications known to alter gastrointestinal motility were excluded. 28 healthy subjects were also recruited. The study group (n = 28) received 90 g per day of ZZ and the control group (n = 28) received 30 mg per day of cisapride for a 1 month period. Conservative therapy was similar in both groups and administered by a person blinded to randomization code. Patients were assessed pre-treatment and post treatment by a blinded interviewer. A constipation visual analogue scale (CVAS) was constructed to evaluate the patients' constipation status. To analyze neuropeptides motilin (MTL), vasoactive intestinal polypeptide (VIP), substance P (SP) levels in plasma, a radioimmunoassay technique was adopted. Data were analyzed for treatment effects using the "t" test. RESULT: Base line characteristics were similar in both treatment groups. There were significant effects on symptom relief, increases in serum SP level, and decreases in VIP level in both groups (P < 0.05). ZZ was significantly more effective than cisapride in ameliorating patients' subjective symptoms of defecating and stool frequency, and increasing serum SP level (P < 0.05). CONCLUSION: ZZ is beneficial in constipating DM via increasing SP levels and decreasing VIP levels of serum. PMID- 19294864 TI - [Research on nature of Chinese materia medica in view of genetic relationship]. AB - In this paper, based on analyzing the status quo and key problem of research on nature of Chinese materia medica, we propose the thinking of research on nature of Chinese materia medica in view of genetic relationship. Namely, according to the cue of documents arrangement and data excavation, in certern scope of genetic relationship, difinite the common pharmaco-effect of the same nature of drug, and analyze the chemical composition correlated the nature of drug, further more, reveal the related regularity of nature of Chinese materia medica-pharmaco-effect chemical composition. Linking the nature of Chinese materia medica, pharmaco effect and chemical composition will provide original thinking and view angle to break through the key problem of research on nature of Chinese materia medica. PMID- 19294865 TI - [Textural research on Zibei Xiqi in Bencao Gangmu Shiyi (a supplement to compendium of materia medica)]. AB - OBJECTIVE: To research the origin of Zibei Xiqi which included in Volume 3 of < Bencao Gangmu Shiyi > (A supplement to the compendium of materia medica). METHOD: The origin of Zibei Xiqi was confirmed with textural, botanical characteristics research and drug name, therapeutic effects and so on. RESULT: The origin of Zibei Xiqi was confirmed. CONCLUSION: The Zibei Xiqi which is recorded in < Bencao Gangmu Shiyi > should be the immature herbs of Shuang Hudie [Tripterospermum chinense, Gentianaceae], and suggest to correct the original records of the drug name. PMID- 19294866 TI - [Suggestions for prevention of adverse reactions after intravasal administration of iodinated contrast media]. AB - Iodinated contrast media are widely used in computed tomography and angiography. Adverse reactions such as contrast-medium induced nephropathy (CIN), anaphylactoid reactions and iodine-induced thyrotoxicosis are associated with intravasal administration of contrast agents. Iodinated contrast agents are generally considered to be safe, but in rare cases they can cause severe life threatening situations. In this review we present an overview about the incidence, pathways, and risk factors of adverse reactions. Simple schemes including hydration protocols for prevention of CIN, medication for prophylaxis of iodine-induced thyrotoxicosis with thyreostatics and anaphylactoid reactions with histamine antagonists and corticosteroids are suggested. PMID- 19294867 TI - [Diagnosis of fungal pneumonia in the thoracic CT]. AB - The importance of fungal infection of the lung in immunocompromised patients has increased substantially during the last decades. Numerically the most patients are those with neutropenia, e.g., patients with malignancies or solid organ and stem cell transplantation, chemotherapy, corticosteroid use and HIV infection. Although fungal infections can occur in immunocompetent patients, their frequency in this population is rare. The clinical symptoms such as fever accompanied with non-productive cough are unspecific. In some patients progression to hypoxemia and dyspnea may occur rapidly. In spite of improved antifungal therapy morbidity and mortality of these infections are still high. Therefore an early and non invasive diagnosis is very important. That is why CT and even better High Resolution-CT (HR-CT) is a very important modality in examining immunocompromised patients with a probability of fungal infection. CT is everywhere available and, as a non-invasive method, able to give the relevant diagnose efficiently. This paper should give an overview about the radiologic findings and possible differential diagnosis of diverse pulmonary fungal infections in CT. Pneumonias caused by Aspergillus, Cryptococcus, Candida, Histoplasma, Mucor and Geotrichum capitatum are illustrated. PMID- 19294868 TI - [Differential diagnosis of focal lesions of the kidney in CT and MRT]. AB - The great majority of renal masses are found incidentally as a result of the use of ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI). If ultrasonography is not diagnostic CT or MRI should be initiated to differentiate lesions of the kidney that need surgical intervention from those that do not and from those that need follow-up examinations. Cystic renal masses are characterized by using the Bosniak classification, including category IIF. In solid Lesions of the kidney first non-surgical lesions as well as lymphoma, renal infarction and nephritis should be excluded. Identifying fatty components in renal lesions is very important because in angiomyolipoma they are almost always present. CT and MRI are exellent for tumor detection. Careful evaluation of imaging finding combined with the patient's history should assist the radiologist in making the proper diagnosis or recommending the appropriate treatment in most cases. This article provides a review about renal masses, the imaging methods for their evaluation and their characteristic features at CT and MR imaging. Different lesions are demonstrated like xantogranulomatous pyelonephritis, acute pyelonephritis, renal infarction, lymphoma, angiomyolipoma, renal oncocytoma, cystic lesion and polycystic disease the kidney, echinococcosis, renal cystadenoma, metastases, renal cell carcinoma (RCC), and multiple bilateral RCC in patients with Hippel-Lindau-Syndrome. This article should help to differentiate complex cystic lesions of the kidney by using the Bosniak classification, especially Bosniak Category IIF. Solid masses should be characterized and the major question to be answered is whether the mass represents a surgical or nonsurgical lesion or if follow-up studies are necessary. PMID- 19294869 TI - [The von Meyenburg complex]. AB - The von Meyenburg complex (VMC) describes bile duct hamartomas and presents a rare, benign incidental finding in liver imaging. We report on a 61-year-old man, who was referred for a follow up study 14 years after remission of Hodgkin's disease. Computed tomography (CT) revealed multiple hypodense lesions throughout the liver, primary suggesting recurrent Hodgkin's disease. Previous CT examinations, which were obtained at a later date, showed those Lesions in identical distribution and morphology over the years, leading to diagnosis of multiple bile duct hamartomas (VMC). Making imaging-based diagnosis of VMC including ultrasound, CT and magnetic resonance imaging is a challenging task for the radiologist. Based on literature research findings, the impact of different modalities in the diagnostic work-up of VMC is discussed. PMID- 19294870 TI - [Hypoxic injury of all deep nuclei of the brain--a case report from computed tomography]. AB - Though being inferior to magnetic resonance imaging, computed tomography (CT) of the brain is the most frequently applied imaging modality in the diagnostic workup of acute cerebral Ischaemia. We report on a case of a comatose 53-year-old man who was brought to the emergency room after cardiopulmonary resuscitation. The CT of the brain showed a diffuse brain oedema with an explicit hypodense demarcation of all deep nuclei. PMID- 19294871 TI - Extraosseous manifestation of multiple myeloma with unusual appearance in computed tomography--case report. AB - Extramedullary Localizations at diagnosis or during the course of multiple myeloma are rare. We report on a 70 year old patient, presenting multiple hypoechoic liver lesions during an ultrasound examination. The following contrast enhanced computed tomography demonstrated hypodense liver Lesions with slight contrast enhancement and hyperdense polypoid masses in the wall of the gall bladder as well as a small pericostal tumor. A punch biopsy of the liver and immunohistochemical studies confirmed the diagnosis of extramedullary multiple myeloma. In a follow-up CT five weeks later the liver lesions and the pericostal tumor clearly showed progress, the masses in the gall bladder had developed into a concentric wall-thickening. Additionally, polypoid contrast-enhancing masses in the gastric wall became apparent as well as a hypodense lesion in the spleen. Radiologists should be aware that multiple myeloma can on rare occasions present as hypodense nodules in the liver or new masses in other organs in CT. Because of the morphologic similarity to metastatic disease, a biopsy may be necessary for definitive diagnosis. PMID- 19294872 TI - [Clinical value of dual source computerized tomography in primary diagnosis of coronary heart disease]. AB - Dual-source-CT-technology (DSCT) improves temporal resolution of cardiac computed tomography to 83 ms per heart-phase. In this study, the clinical performance of this new method is evaluated. MATERIALS AND METHODS: In fifty patients (33 male, 17 female; age 50 +/- 13 years) with suspected coronary heart disease, CT angiography (slice thickness 0.75 mm, contrast-agent 60-80 ml iomeprol) was performed with a Somatom Definition scanner. Based on the coronary 15-segment model of the AHA, scores for image quality and lumen reduction were established to enable the observer, to give recommendations for further therapy. RESULTS: Out of 750 possible AHA-segments, 655 were depicted (87.3%). 591 segments (90.2%) were assessed without any limitation of quality, 49 (7.5%) segments showed moderate, and 15 (2.3%) segments severe limitation in image quality. 508 (77.6%) segments were without pathological findings, 92 (14.0%) segments had minimal atherosclerotic lesions, 42 (6.4%) segments suffered from stenoses with lumen reduction less than 70%, and 13 (2.0%) showed significant stenoses of more than 70%. In 31 patients (62%), coronary heart disease was ruled out by CT angiography without any need for further non-invasive or invasive diagnostics. 8 patients (16%) underwent stress-testing for ischemia. In 11 (22%) patients coronary angiography was recommended, and DSCT findings were confirmed in 10 cases. Only one LCx stenosis was overestimated in DSCT. CONCLUSION: Contrast-enhanced DSCT is a powerful tool in diagnosis of coronary heart disease. 98% of coronary segments could be assessed in diagnostic quality, and at least 90% of haemodynamically significant coronary stenoses were detected. PMID- 19294873 TI - Endovascular treatment of traumatic high flow dural arterio-venous fistula involving the middle meningeal artery and facial veins. AB - Dural arterio-venous fistulas are rare lesions. Traumatic dural AV-fistulas in particular, are very rare complications of head and neck trauma. We report on a 59-year old man with a severe facial and head trauma. Initial computed tomography demonstrated a left frontal subdural hematoma, leading to progressive midline shift. Additionally, a complex skull base fracture was present, involving the frontal, temporal and sphenoid bone. A few days after the accident the patient reported a left pulsatile tinnitus. Subsequent selective carotid angiography revealed a high flow dural arterio-venous fistula involving the middle meningeal artery and facial veins. Complete endovascular embolization was performed by a transarterial approach using microparticles and an electrolytically detachable coil resulting in an immediate disappearance of the tinnitus. Remarkably, this case nicely illustrates possible "dangerous" collateral circulation to the ophthalmic artery-a feature that should be kept in mind during endovascular treatment of this entity. PMID- 19294874 TI - [Cosmetic results after breast conserving carcinoma treatment in patients with intramammarian seromas]. AB - INTRODUCTION: There were 373 patients irradiated after breast conserving carcinoma treatment. A planning computed tomography revealed in 97 of these patients seromas and tissue defects exceeding 2cm in diameter. The cosmetic results in those patients and the impact of seromas herein had to be evaluated. PATIENTS AND METHODS: Mean age was 59 years. A quadrant resection was performed in 17.5 percent of the patients, a segmental resection in 27.8 percent and a tumour excision in 54.6 percent. Radiation therapy was applied with the Linear accelerator and 6 MeV photons up to a total dose in the residual breast of 50 Gy followed by a boost dose to the former tumour bed of 10 Gy. A distinct evaluation and documentation of therapy related side effects and the resulting cosmesis was done in 51 patients. RESULTS: In all the examined seroma patients there were moderate acute skin reactions grade 1 to 3. As late effects in 82.3 percent scar indurations were noticed. At the skin 51 percent showed enhanced pigmentation, 68.6 percent atrophia and only 11 percent teleangiectasia. Subcutaneous fibrosis occurred in 56,9 percent of the patients, 78.4 percent of the women had breast asymmetries. In 41.2 percent there were a lymphedema subcutaneously, in 72.5 percent impaired sensibility. The overall cosmetic result documented with a 5 point score was "very good" (score 1) in 19.6 percent and "good" (score 2) in 33.3 percent, 37.3 percent were "satisfactory" (score 3) and 9.8 percent "bad" (score 4) respectively. No "very bad" results (score 5) were seen. CONCLUSIONS: The cosmetic results in the examined group of seroma and hematoma patients were inferior to those reported in the literature. We conclude that postoperative seroma and hematoma have an adverse effect on the resulting cosmesis and that their frequency and extent have to be reduced in future by the responsible surgeons. PMID- 19294875 TI - [Popliteal artery entrapment syndrome (PAES)--differential diagnosis of claudication intermittens]. PMID- 19294876 TI - [Refined staging in Hilar bronchial neoplasms with ECG-gated multislice-CT---case report]. AB - Equivocal initial CT-based staging in 2 patients with hilar bronchial neoplasms was reassessed with retrospective ECG-gated Multislice-CT and optimized examination parameters prior to definition of treatment. An initially suspected irresectable T4 tumor with mediastinal infiltration was downstaged to T2 in one case, while tumor infiltration into the left atrium could be confirmed in the other case. In doubtful conditions, ECG-gated multislice CT with optimized examination parameters may be helpful for refined staging in patients with hilar bronchial neoplasms, thus possibly influencing treatment strategies. PMID- 19294877 TI - [Spontaneously reversible portal vein thrombosis complicating acute pancreatits- computed tomographic findings]. AB - Portal vein thrombosis complicating acute pancreatitis is more often diagnosed today due to the improved imaging techniques (computed tomography, ultrasound, nmr). Nevertheless the outcome of recent portal vein thrombosis is ill-known. We report on the computed tomographic findings and clinical course of portal vein thrombosis in two patients suffering from acute pancreatitis. Both patients showed spontaneous recanalization of the thrombosis. PMID- 19294878 TI - [Weight bearing upright magnetic resonance imaging of pointe dancing]. AB - Weight-bearing upright Magnetic resonance imaging (MRI) "en pointe" is the ideal tool for research in the field of functional anatomy of pointe dancing and pointe dancing related medicine. PMID- 19294879 TI - Oh data, where are ye? Using administrative databases in epilepsy research. PMID- 19294880 TI - Managing unruptured intracranial aneurysms. PMID- 19294881 TI - Reflections on aspects of medical progress 1944-2008--part 1. The launch of a journey lasting 65 years: personal observations made from a front-row seat at the greatest show on earth. PMID- 19294882 TI - The role of polyamine metabolism in neuronal injury following cerebral ischemia. AB - Stroke is a leading cause of morbidity and mortality in the US, with secondary damage following the initial insult contributing significantly to overall poor outcome. Prior investigations have shown that the metabolism of certain polyamines such as spermine, spermidine, and putrescine are elevated in ischemic parenchyma, resulting in an increase in their metabolite concentration. Polyamine metabolites tend to be cytotoxic, leading to neuronal injury in the penumbra following stroke and expansion of the area of infarcted tissue. Although the precise mechanism is unclear, the presence of reactive aldehydes produced through polyamine metabolism, such as 3-aminopropanal and acrolein, have been shown to correlate with the incidence of cerebral vasospasm, disruption of oxidative metabolism and mitochondrial functioning, and disturbance of cellular calcium ion channels. Regulation of the polyamine metabolic pathway, therefore, may have the potential to limit injury following cerebral ischemia. To this end, we review this pathway in detail with an emphasis on clinical applicability. PMID- 19294883 TI - Meta-analysis of the ACE gene polymorphism in cerebral infarction. AB - BACKGROUND: The angiotensin-1 converting enzyme (ACE) gene is known to have two polymorphic alleles insertion/deletion (I/D). People with the DD genotype have been shown to be at greater risk of cerebral infarction, but only in some studies. Identification of cerebral infarction susceptibility genes and quantification of associated risks have been hampered by conflicting results from underpowered case-control studies. This meta-analysis was made to look specifically into the genetics of cerebral infarction among Han Chinese population. METHODS: Genetic associations studies published from January 1, 1990 to December 30, 2007 were collected from databases of MEDLINE, EMBASE, CBM and CNKI. Data were extracted using standardised forms and pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. RESULTS: Twenty-nine original case-control studies of Han Chinese population, comprising 3654 patients with cerebral infarction and 3058 controls were included in the meta-analysis. Using the random effects model, the pooled ORs of ACE DD genotype VS ID+ II was 1.91 (95% CI 1.56 to 2.34, P<0.00001). CONCLUSIONS: These data suggest that the ACE DD genotype may be a risk factor for cerebral infarction in Han Chinese population. A large scale case-control study is needed to clarify the functional effect of the polymorphism of the ACE I/D gene in the pathogenesis of cerebral infarction in Han Chinese population. PMID- 19294884 TI - Complex I and ATP content deficiency in lymphocytes from Friedreich's ataxia. AB - BACKGROUND: Friedreich's ataxia (FRDA) is an inherited recessive disorder characterized by progressive neurological disability and heart abnormalities. A deficiency in the protein frataxin causes this disease. Frataxin deficiency leads to progressive iron accumulation in mitochondria, excessive free radical production and dysfunction of respiratory chain complexes. The expansion (GAA) repeat in the first intron causes decreased frataxin expression by interfering with transcription. METHODS: Activity of mitochondrial respiratory chain complex I (measured as NADH ferricyanide reductase) and intracellular ATP measurement was performed on lymphocyte of FRDA patients (n=12) and control subjects (n=25). RESULTS: Our findings showed that complex I activity and intracellular ATP were significantly reduced (P=0.001) in patients compared with controls and we found strong correlation between complex I activity and intracellular ATP content in FRDA patients (r=0.93; P<0.002). 8.6 and 9.0 kb deletion in mtDNA was detected in 9 patients out of 12 (75%) by multiplex polymerase chain reaction (PCR) and Southern blot analysis. CONCLUSIONS: This study suggested that a biochemical defect in complex I activity and ATP production, which may be due to iron accumulation in mitochondria, can be involved in age of onset of FRDA. PMID- 19294885 TI - Potentially life-threatening ictal bradycardia in intractable epilepsy. AB - BACKGROUND: Recently a high prevalence of asystole was found in individuals with refractory epilepsy. OBJECTIVE: To measure the prevalence and characterize the pattern of ictal bradycardia (IB) in our cohort. METHOD: We analyzed ictal heart rate (HR) and seizure localization in 69 consecutive individuals with intractable epilepsy. Ictal bradycardia was defined by HR less than 60 beat per min (bpm) and a fall of more than 30 bpm. RESULTS: Ictal bradycardia was observed in 1.9% of seizures and in five individuals (7.2%), who presented IB in 5.7% of their seizures. Four of them had also relative IB (a decrease of HR of more than l0 bpm) in the majority of their seizures. Four additional individuals showed relative IB. CONCLUSION: Ictal bradycardia occurs more frequently than previously thought in individuals with refractory epilepsy. Recognition of individuals with small to moderate ictal HR decrease may help to identify those at greatest risk for asystole. PMID- 19294886 TI - Period prevalence of epilepsy in children in BC: a population-based study. AB - BACKGROUND: Most estimates of the prevalence of seizure disorders in Canada derive from national surveys which differ in sampling and case-finding methods. This study used health care utilization data to make a population-based estimate of the prevalence of epileptic seizures and of epilepsy in children in British Columbia (BC). METHODS: All BC residents between 0-19 years-of-age in 2002-3 enrolled in the Medical Services Plan were included. Epileptic seizures were defined using ICD-9 codes; health care utilization data was obtained from BC Linked Health Database. The period prevalence of epileptic seizures and of epilepsy was determined by age, urban/rural region and socioeconomic status. RESULTS: 8,125 of 1,013,816 children were identified as having an epileptic seizure of which 5621 were classified as epilepsy--5.5 per 1000 children (95% CI: 5.4-5.7). The prevalence of epilepsy in infants and preschoolers was higher than that reported in the literature. A higher prevalence of epilepsy was observed also among those with low socioeconomic status. A higher prevalence of epilepsy was observed in those health regions with a higher proportion of First Nations and a lower prevalence was observed in health regions with a higher proportion of visible minorities. CONCLUSIONS: Age-specific prevalence rates in BC children for epilepsy, determined from population-based administrative records, were similar to published data except in children under five years. We found a gradient of increased prevalence with decreased level of income. Prevalence rates based on utilization data have the potential to guide program planning for children with epileptic seizures. PMID- 19294887 TI - Regional variability in the use of CT for patients with suspected mild traumatic brain injury. AB - OBJECTIVE: To investigate the use of computed tomography (CT) scans in patients with suspected acute mild traumatic brain injury (mTBI) presenting to emergency departments. METHOD: 850 potential mTBI cases were identified through reviews of three months of health records from nine selected emergency departments across the province of Ontario. Records for review were selected using the International Classification of Disease, 9th revision, Clinical Modification codes and Injury codes. RESULTS: Patients who received head CT were significantly older (p<0.01), had documented loss-of-consciousness (LOC) &/or Post-Traumatic Amnesia (PTA) (p<0.001), documented nausea (p<0.01), documented vomiting (p<0.001), abnormal neurological exam results (p<0.01), had visited an urban center (p<0.001), and/or arrived by ambulance (p<0.001). The significant predictors of CT scan prescription (in a forward stepwise logistic regression) were urban location of hospital (OR=5.14; p<0.001), LOC &/or PTA (OR=4.83; p< or =0.001), vomiting (OR=2.56; p< or =0.01), arrival by ambulance (OR=2.15; p< or =0.001), nausea (OR=1.92; p< or =0.02) and older age (OR=1.02; p< or =0.01). CONCLUSION: These data extend our knowledge regarding the use of CT during acute diagnosis and management of suspected mTBI patients. In addition to confirming previously reported risk factors of intracranial complication, geographical location of hospital and arrival mode were found to be significant predictors of CT use. The results suggest that the management patterns for acute mTBI are inconsistent. The implications of this are discussed. PMID- 19294888 TI - Neuromonitoring during surgery for paediatric spinal deformity in Canada (2007). AB - BACKGROUND: Neuromonitoring during paediatric (and adult) spinal deformity surgery helps to reduce the risk of both permanent and short term neurological damage. A shortage of neurophysiologists and technicians limits the availability of this service. Not all surgeons believe neuromonitoring offers neuroprotection during spinal surgery. This study aimed to document the degree to which paediatric patients undergoing spinal deformity correction surgery have their spinal cord function monitored. METHODS: A questionnaire was sent electronically to all of the surgical members of the Canadian Paediatric Spinal Deformity Study Group. RESULTS: Results were received from 9/9 centres indicating that monitoring was performed in 7/9 centres, with one further centre awaiting staffing. Whilst half of those centres that do monitor only monitor sensory and motor evoked potentials, the remaining centres also use EMG and EEG to assess the state of the patient intraoperatively. CONCLUSIONS: Despite a shortage of staff, most paediatric spinal deformity surgeons in Canada who wish to, are able to neurophysiologically monitor their surgical cases. Neuromonitoring appears to be becoming a standard of care, at least for paediatric spinal deformity surgery. There is an urgent need for the establishment of national standards for both technologists and interpreters, as well as training programmes for both these groups. PMID- 19294889 TI - Group education with personal rehabilitation for idiopathic Parkinson's disease. AB - OBJECTIVE: To evaluate the effect of a group education program with personal rehabilitation for idiopathic Parkinson's disease (IPD). METHODS: A single-blind, randomized, controlled clinical trial, with a pre-test/post-test quasi experimental design. Forty-four stable, non-demented patients with IPD were randomly assigned to an intervention group (n=23) and control group (n=21). Three group lectures on health education specific to IPD were delivered to the intervention group. The theme of lectures was "Not a lonely journey, as your friend, we---experts specializing in IPD---would love to join with you". Three domains of the management of IPD were mentioned: "How to manage your routine Meal, Moving and Mood after suffering from IPD" Each lecture was scheduled for 45 minutes (excluding breaks), and designed as an interactive dialogue rather than a traditional lecture. To strengthen the impact of the lectures, relevant information was published on a website: http://www.parkinsonism.cn/. Following the group lecture, individualized and tailored rehabilitation made up of 24 half hour sessions over eight weeks were provided for the intervention group. RESULTS: The program ran smoothly and IPD patients were satisfied with this program. After only eight weeks, the health-related quality of life (HRQOL) in the intervention group improved significantly (p<0.001). Patients and their caregivers in this group also reported their mood elevations following the program. CONCLUSION: Group education combined with personal rehabilitation program appears to be a beneficial and practical intervention, not only complementing the medical treatment for IPD patients but also meeting the growing demand for long-term care. PMID- 19294890 TI - Bulbar signs in normal population. AB - BACKGROUND AND OBJECTIVES: There is lack of published data on bulbar signs among the healthy population. This study aims to determine the range of normality of bulbar signs particularly among the elderly. METHODS: Systemic examination of bulbar signs was carried out according to a predetermined protocol on a cohort of young and elderly healthy subjects. RESULTS: A total of 206 subjects were recruited in the study, 104 young adults with mean age of 20 years, and 102 elderly with mean age of 73 years. Uvula deviation was seen in 28 (26.9%) young subjects and 22 (21.6%) elderly. Irregular tongue border was seen in 17 subjects, unilateral in 4 subjects. Fourteen (6.8%) subjects had deviation on tongue protrusion. Occasional tremor of tongue on protrusion is common in both young and old. Persistent (severe) tongue tremor on protrusion was seen in 18.6% of the elderly, and 4.8% of the young. None of the subjects had tremor of tongue at rest. In gag reflex, absence of gagging response was common in elderly, seen in two thirds of the subjects on stimulation of the posterior pharyngeal wall. However, all the subjects had uvular movement. Habituation or suppression of gagging response was seen in close to 90% of young males. CONCLUSION: There is wide range of normality in bulbar signs in normal population, particularly among the elderly. PMID- 19294891 TI - Superficial anastomotic veins: neurosurgical view depending on 251 craniotomies. AB - BACKGROUND: Superficial anastomotic veins (SAVs) have been studied extensively but little attention has been paid to clinical studies. The aim of this study is to provide variations in the drainage patterns of SAVs depending on the intraoperative findings. METHODS: A total of 251 craniotomies due to intractable temporal lobe epilepsy were performed between 1972 and 1987 at the Montreal Neurological Institute. The courses of the three largest SAVs including the vein of Trolard (VT), vein of Labbe (VL) and superficial Sylvian vein (SSV) were studied. RESULTS: All three veins showed variable courses. The most common predominant vein was the combination of the VL + SSV. The VT and VL were frequently coursed at the level of the central vein and middle temporal vein, respectively. On the right hemisphere the SSV was the predominant type while the VL tended to be predominant on the left hemisphere. A combination of VL and SSV was predominant in patients with right and/or left hemispheric dominance. CONCLUSIONS: The SAVs showed considerable variation in their courses and it is difficult to define an exact pattern although some courses showed constant directions. Attention should be paid not to damage these veins since in a considerable number of cases a single dominant vein may be responsible for draining a majority of the lateral surface of cerebral hemisphere. PMID- 19294892 TI - Malformations of the fetal dural sinuses. AB - BACKGROUND: Dural sinus malformation (DSM) is a term used to describe congenital vascular malformations characterized by massive dilation of one or more dural sinuses: these dilatations are typically associated with arteriovenous shunts. Such malformations can present antenatally but their early natural history and anatomy is poorly defined. METHODS: We reviewed five years of autopsy experience and retrieved three primary vascular malformations of the fetal dural sinuses with ultrasound, magnetic resonance imaging (MRI) and post-mortem correlation. RESULTS: Fetal ultrasound and MRI obtained between 19 and 23 weeks gestational age demonstrated in all cases dilation of the dural sinuses. In two cases vascular thromboses were present in either the dilated dural sinus (one of three) or the associated arteriovenous fistula (one of three). All cases were autopsied at 22-23 weeks gestational age. In one there was imaging and autopsy evidence of remodeling of the dural sinuses associated with a pial arteriovenous fistula. In two cases, no arteriovenous malformation was identified on initial imaging, but only became evident at autopsy. One case showed morphological overlap with vein of Galen aneurysmal malformation, with a midline arteriovenous shunt and vein of Galen ectasia. The other demonstrated a perisylvian dural arteriovenous fistula. CONCLUSION: In utero thrombosis of feeding vascular malformations or of the dural sinus malformation may be prominent. The early in utero developmental trajectory of dural sinus malformation (DSM) is poorly defined and deserves further study. PMID- 19294893 TI - Trigeminal neuralgia: outcomes after gamma knife radiosurgery. AB - BACKGROUND: Trigeminal neuralgia (TN) often remains difficult to treat despite multiple available medications, and can severely impact on the quality of life of affected patients. Gamma knife radiosurgery has recently emerged as a minimally invasive alternative to surgery for patients suffering from drug-resistant TN. The goal of this study was to report the short-term efficacy of gamma knife radiosurgery for TN and assess its impact on the quality of life of patients treated in the first 18 months of our experience. METHODS: Patients with medically-refractory TN or with unacceptable drug side effects were considered for radiosurgery. A maximum dose of 80 Gy was administered to the affected nerve using a single 4-mm isocenter. Follow-up assessments were made at 2,4 and 6 months, with evaluation of pain relief, drug reduction and quality of life. Factors impacting treatment response were assessed using Cox regression analysis. RESULTS: A total of 67 patients were treated. Significant pain relief was seen in 77.6% of patients, including 32.6% who became pain-free. Patients were able to discontinue all medications in 34.3% or reduce drug intake by more than 50% in an additional 28.4% of cases. No variable was found to predict pain relief although older age (>66 years) approached statistical significance. Sensory side effects were seen in 14.9% of patients. Quality of life improved in the majority of patients after radiosurgery. CONCLUSIONS: Gamma knife radiosurgery is a safe and effective management alternative for trigeminal neuralgia, providing good or excellent pain relief and improvement in quality of life in the majority of patients with few side effects. PMID- 19294894 TI - TMS neuro-cardiovascular coupling in vascular compression cranial neuropathy. AB - BACKGROUND: Neurovascular compression (NVC) may cause cranial mononeuropathy but lacks a definitive diagnostic investigation. We hypothesized that the arterial pressure wave (APW) would interact at the neurovascular interface in NVC to inhibit transmission of transcranial magnetic stimulation (TMS) stimuli to affected muscles. METHODS: We report a novel neurophysiological method coupling cardiovascular physiology with TMS. The electrocardiogram (ECG) and arterial pressure wave (APW) were coupled to triggering of cortical TMS in a patient with NVC-induced spinal accessory (CNXI) mononeuropathy. Outcome measures included motor evoked potential (MEP) amplitudes and firing probabilities of normal and affected trapezieus (TPZ). Values at intervals in proximity to the APW (40/80/120/160 ms) were compared to baseline (800 ms) using ANOVA and student t test. RESULTS: Electrocardiogram triggered TMS of CNXI pathways with 100% reliability. MEP amplitudes were decreased in proximity to the APW, particularly at 120 ms (0.21-0.04 mV versus 0.39 +/- 0.10 mV, p=0.003). TPZ firing probabilities were similarly inhibited (43.8% versus 88.2%, p=0.009). No effect of APW proximity was observed on the unaffected side (p=0.868). Procedures were well tolerated. CONCLUSIONS: Vascular compression causes CNXI mononeuropathy. Transcranial magnetic stimulation-cardiovascular coupling may evaluate neurovascular junction interactions and non-invasively diagnose NVC. PMID- 19294895 TI - Optimization of visual evoked potential (VEP) recording systems. AB - OBJECTIVE: To explore the influence of environmental conditions on pattern visual evoked potential (VEP) recordings. METHODS: Fourteen subjects with no known ocular pathology were recruited for the study. In an attempt to optimize the recording conditions, VEP recordings were performed in both the seated and recumbent positions. Comparisons were made between recordings using either LCD or CRT displays and recordings obtained in silence or with quiet background music. Paired recordings (in which only one variable was changed) were analyzed for changes in P100 latency, RMS noise, and variability. RESULTS: Baseline RMS noise demonstrated a significant decrease in the variability during the first 50msec accompanied by a 73% decrease in recording time for recumbent position when compared to the seated position (p<0.05). Visual evoked potentials recorded using LCD monitors demonstrated a significant increase in the P100 latency when compared to CRT recordings in the same subjects. The addition of background music did not affect the amount of RMS noise during the first 50msec of the recordings. CONCLUSION: This study demonstrates that the use of the recumbent position increases patient comfort and improves the signal to noise ratio. In contrast, the addition of background music to relax the patient did not improve the recording signal. Furthermore, the study illustrates the importance of avoiding low-contrast visual stimulation patterns obtained with LCD as they lead to higher latencies resulting in false positive recordings. These findings are important when establishing or modifying a pattern VEP recording protocol. PMID- 19294896 TI - Cerebral angiography and diagnosis of CNS vasculitis. PMID- 19294897 TI - Reversible parinaud syndrome following intraventricular thrombolysis. PMID- 19294898 TI - Systemic lupus erythematosus and right leg weakness. PMID- 19294899 TI - Intraventricular neurocysticercosis managed with albendazole and dexamethasone. PMID- 19294900 TI - Hypotonia and infantile spasms: a new phenotype of coenzyme Q10 deficiency? PMID- 19294901 TI - Grisel syndrome in adult patients. Report of two cases and review of the literature. PMID- 19294902 TI - Congenital dermal sinus and cyst in adulthood. PMID- 19294903 TI - Cryptococcal choroid plexitis an uncommon fungal disease. Case report and review. PMID- 19294904 TI - Silent somatotroph adenoma of the pituitary in an adolescent. PMID- 19294905 TI - Withdrawal of care. PMID- 19294906 TI - Modeling misidentification errors in capture-recapture studies using photographic identification of evolving marks. AB - Misidentification of animals is potentially important when naturally existing features (natural tags) are used to identify individual animals in a capture recapture study. Photographic identification (photoID) typically uses photographic images of animals' naturally existing features as tags (photographic tags) and is subject to two main causes of identification errors: those related to quality of photographs (non-evolving natural tags) and those related to changes in natural marks (evolving natural tags). The conventional methods for analysis of capture-recapture data do not account for identification errors, and to do so requires a detailed understanding of the misidentification mechanism. Focusing on the situation where errors are due to evolving natural tags, we propose a misidentification mechanism and outline a framework for modeling the effect of misidentification in closed population studies. We introduce methods for estimating population size based on this model. Using a simulation study, we show that conventional estimators can seriously overestimate population size when errors due to misidentification are ignored, and that, in comparison, our new estimators have better properties except in cases with low capture probabilities (< 0.2) or low misidentification rates (< 2.5%). PMID- 19294907 TI - Perturbation analysis for patch occupancy dynamics. AB - Perturbation analysis is a powerful tool to study population and community dynamics. This article describes expressions for sensitivity metrics reflecting changes in equilibrium occupancy resulting from small changes in the vital rates of patch occupancy dynamics (i.e., probabilities of local patch colonization and extinction). We illustrate our approach with a case study of occupancy dynamics of Golden Eagle (Aquila chrysaetos) nesting territories. Examination of the hypothesis of system equilibrium suggests that the system satisfies equilibrium conditions. Estimates of vital rates obtained using patch occupancy models are used to estimate equilibrium patch occupancy of eagles. We then compute estimates of sensitivity metrics and discuss their implications for eagle population ecology and management. Finally, we discuss the intuition underlying our sensitivity metrics and then provide examples of ecological questions that can be addressed using perturbation analyses. For instance, the sensitivity metrics lead to predictions about the relative importance of local colonization and local extinction probabilities in influencing equilibrium occupancy for rare and common species. PMID- 19294908 TI - Nesting lizards (Bassiana duperreyi) compensate partly, but not completely, for climate change. AB - Species in which ambient temperatures directly determine offspring sex may be at particular risk as global climates change. Whether or not climate change affects sex ratio depends upon the effectiveness of buffering mechanisms that link ambient regimes to actual nest temperatures. For example, females may simply lay nests earlier in the season, or in more shaded areas, such that incubation thermal regimes are unchanged despite massive ambient fluctuation. Based on eight years of monitoring nests over a 10-year period in the field at an alpine site in southeastern Australia, we show that, even though lizards (Bassiana duperreyi, Scincidae) have adjusted both nest depth and seasonal timing of oviposition in response to rising ambient temperatures, they have been unable to compensate entirely for climate change. That inability stems from the fact that the seasonal progression of soil temperatures, and thus, the degree to which thermal regimes at the time of laying predict subsequent conditions during incubation, also has shifted with climate change. As a result, mean incubation temperatures in natural nests now have crossed the thermal threshold at which incubation temperature directly affects offspring sex in this population. PMID- 19294909 TI - Behavioral adjustments of African herbivores to predation risk by lions: spatiotemporal variations influence habitat use. AB - Predators may influence their prey populations not only through direct lethal effects, but also through indirect behavioral changes. Here, we combined spatiotemporal fine-scale data from GPS radio collars on lions with habitat use information on 11 African herbivores in Hwange National Park (Zimbabwe) to test whether the risk of predation by lions influenced the distribution of herbivores in the landscape. Effects of long-term risk of predation (likelihood of lion presence calculated over four months) and short-term risk of predation (actual presence of lions in the vicinity in the preceding 24 hours) were contrasted. The long-term risk of predation by lions appeared to influence the distributions of all browsers across the landscape, but not of grazers. This result strongly suggests that browsers and grazers, which face different ecological constraints, are influenced at different spatial and temporal scales in the variation of the risk of predation by lions. The results also show that all herbivores tend to use more open habitats preferentially when lions are in their vicinity, probably an effective anti-predator behavior against such an ambush predator. Behaviorally induced effects of lions may therefore contribute significantly to structuring African herbivore communities, and hence possibly their effects on savanna ecosystems. PMID- 19294910 TI - Demographic trade-offs in a neutral model explain death-rate--abundance-rank relationship. AB - The neutral theory of biodiversity has been criticized for its neglect of species differences. Yet it is much less heeded that S. P. Hubbell's definition of neutrality allows species to differ in their birth and death rates as long as they have an equal per capita fitness. Using the lottery model of competition we find that fitness equalization through birth-death trade-offs can make species coexist longer than expected for demographically identical species, whereas the probability of monodominance for a species under zero-sum neutral dynamics is equal to its initial relative abundance. Furthermore, if newly arising species in a community survive preferentially they are more likely to slip through the quagmire of rareness, thus creating a strong selective bias favoring their community membership. On the other hand, high-mortality species, once having gained a footing in the community, are more likely to become abundant due to their compensatory high birth rates. This unexpected result explains why a positive association between species abundance and per capita death rate can be seen in tropical-forest communities. An explicit incorporation of interspecific trade-offs between birth and death into the neutral theory increases the theory's realism as well as its predictive power. PMID- 19294911 TI - Speciation and endemism under the model of island biogeography. AB - Speciation has been considered as a primary process contributing to species diversity, but its contribution to the diversity of local communities has not been fully appreciated. Based on the theory of classic island biogeography, we derived a model for the number of endemic species as a function of the processes of immigration, speciation, and extinction. The model shows that species endemism on an island is proportional to speciation rate but decreases with the sum of immigration and extinction rates (i.e., the species turnover rate). The model predicts that the contribution of immigration to species richness in local communities decreases with time, while the contribution of speciation to local richness increases with time. It further shows that only when the speciation rate is larger than half of the extinction rate can new species added from speciation eventually surpass those added from immigration. We conclude that, although the model leads to an apparent positive relationship between percentage endemism and species diversity on an island, this positive endemics-diversity relationship is not necessarily driven by speciation. PMID- 19294912 TI - Beyond description: the active and effective way to infer processes from spatial patterns. AB - The ecological processes that create spatial patterns have been examined by direct measurement and through measurement of patterns resulting from experimental manipulations. But in many situations, creating experiments and direct measurement of spatial processes can be difficult or impossible. Here, we identify and define a rapidly emerging alternative approach, which we formalize as "space as a surrogate" for unmeasured processes, that is used to maximize inference about ecological processes through the analysis of spatial patterns or spatial residuals alone. This approach requires three elements to be successful: a priori hypotheses, ecological theory and/or knowledge, and precise spatial analysis. We offer new insights into a long-standing debate about process-pattern links in ecology and highlight six recent studies that have successfully examined spatial patterns to understand a diverse array of processes: competition in forest-stand dynamics, dispersal of freshwater fish, movement of American marten, invasion mechanisms of exotic trees, dynamics of natural disturbances, and tropical-plant diversity. Key benefits of using space as a surrogate can be found where experimental manipulation or direct measurements are difficult or expensive to obtain or not possible. We note that, even where experiments can be performed, this procedure may aid in measuring the in situ importance of the processes uncovered through experiments. PMID- 19294913 TI - Stage-structured matrix models for organisms with non-geometric development times. AB - Matrix models have been used to model population growth of organisms for many decades. They are popular because of both their conceptual simplicity and their computational efficiency. For some types of organisms they are relatively accurate in predicting population growth; however, for others the matrix approach does not adequately model growth rate. One of the reasons for the lack of accuracy is that most matrix-based models implicitly assume a specific degree of variability in development times for the organism. Because the variability is implicit, the implied variances are often not verified with experimental data. In this paper, we shall present extensions to the stage-classified matrix models so that organisms with arbitrary means and standard deviations of development times can be modeled. PMID- 19294914 TI - Marketing time predicts naturalization of horticultural plants. AB - Horticulture is an important source of naturalized plants, but our knowledge about naturalization frequencies and potential patterns of naturalization in horticultural plants is limited. We analyzed a unique set of data derived from the detailed sales catalogs (1887-1930) of the most important early Florida, USA, plant nursery (Royal Palm Nursery) to detect naturalization patterns of these horticultural plants in the state. Of the 1903 nonnative species sold by the nursery, 15% naturalized. The probability of plants becoming naturalized increases significantly with the number of years the plants were marketed. Plants that became invasive and naturalized were sold for an average of 19.6 and 14.8 years, respectively, compared to 6.8 years for non-naturalized plants, and the naturalization of plants sold for 30 years or more is 70%. Unexpectedly, plants that were sold earlier were less likely to naturalize than those sold later. The nursery's inexperience, which caused them to grow and market many plants unsuited to Florida during their early period, may account for this pattern. Plants with pantropical distributions and those native to both Africa and Asia were more likely to naturalize (42%), than were plants native to other smaller regions, suggesting that plants with large native ranges were more likely to naturalize. Naturalization percentages also differed according to plant life form, with the most naturalization occurring in aquatic herbs (36.8%) and vines (30.8%). Plants belonging to the families Araceae, Apocynaceae, Convolvulaceae, Moraceae, Oleaceae, and Verbenaceae had higher than expected naturalization. Information theoretic model selection indicated that the number of years a plant was sold, alone or together with the first year a plant was sold, was the strongest predictor of naturalization. Because continued importation and marketing of nonnative horticultural plants will lead to additional plant naturalization and invasion, a comprehensive approach to address this problem, including research to identifyand select noninvasive forms and types of horticultural plants is urgently needed. PMID- 19294915 TI - Including intraspecific variability in functional diversity. AB - Linking species and ecosystems often relies on approaches that consider how the traits exhibited by species affect ecosystem processes. One method is to estimate functional diversity (FD) based on the dispersion of species in functional trait space. Individuals within a species also differ, however, and an unresolved challenge is how to include such intraspecific variability in a measure of functional diversity. Our solution is to extend an existing measure to variation among individuals within species. Here, simulations demonstrate how the new measure behaves relative to one that does not include individual variation. Individual-level FD was less well associated with species richness than species level FD in a single trait dimension, because species differed in their intraspecific variation. However, in multiple trait dimensions, there was a strong association between individual- and species-level FD and richness, because many traits result in a tight relationship between functional diversity and species richness. The correlation between the two FD measures weakened as the amount of intraspecific variability increased. Analyzing natural plant communities we found no relationship between species richness and functional diversity. In these analyses, we did not have to specify the source of intraspecific variation. In fact, the variation was only among individuals. The measure can, however, include differences in the amount of intraspecific variation at different sites, as we demonstrate. Including intraspecific variation should allow a more complete understanding of the processes that link individuals and ecosystems and provide better predictions about the consequences of extinctions for ecosystem processes. PMID- 19294916 TI - Long-term effects of plant diversity and composition on soil nematode communities in model grasslands. AB - An important component of plant-soil feedbacks is how plant species identity anddiversity influence soil organism communities. We examine the effects of grassland plant species growing alone and together up to a richness of 12 species on nematode diversity and feeding group composition, eight years after the establishment of experimental grassland plots at the BIODEPTH site in northern Sweden. This is a substantially longer time than most other experimental studies of plant effects on soil fauna. We address the hypotheses that (la) higher species or functional diversity of plants increases nematode diversity, as well as influences nematode community composition. Alternatively, (1b) individual plant species traits are most important for nematode diversity and community composition. (2) Plant effects on soil organisms will decrease with increasing number of trophic links between plants and soil fauna. Plant species identity was often more important than plant diversity for nematode community composition, supporting hypothesis 1b. There was a weak positive relation between plant and nematode richness;which could be attributed to the presence of the legume Trifolium pratense, but also to some other plant species, suggesting a selection or sampling effect. Several plant species in different functional groups affected nematode community composition. For example, we found that legumes increased bacterial-feeding nematodes, most notably r-selected Rhabditida, while fungal feeding nematodes were enhanced by forbs. Other bacterial feeders and obligate root feeders were positively related to grasses. Plant effects were usually stronger on plant-, bacterial- and fungal-feeding nematodes than on omnivores/predators, which supports hypothesis 2. Our study suggests that plant identity has stronger effects than plant diversity on nematode community composition, but when comparing our results with similar previous studies the effects of particular plant species appear to vary. We also found that more productive plant species affected bacterial-feeding nematodes more than fungal feeders. Moreover, we observed stronger effects the fewer the number of trophic links there were between a nematode feeding group and plants. Although we found clear effects of plants on soil nematodes, these were probably not large enough to result in strong and persistent plant-soil-organism-plant feedback loops. PMID- 19294917 TI - The brighter side of soils: quantum dots track organic nitrogen through fungi and plants. AB - Soil microorganisms mediate many nutrient transformations that are central in terrestrial cycling of carbon and nitrogen. However, uptake of organic nutrients by microorganisms is difficult to study in natural systems. We assessed quantum dots (fluorescent nanoscale semiconductors) as a new tool to observe uptake and translocation of organic nitrogen by fungi and plants. We conjugated quantum dots to the amino groups of glycine, arginine, and chitosan and incubated them with Penicillium fungi (a saprotroph) and annual bluegrass (Poa annua) inoculated with arbuscular mycorrhizal fungi. As experimental controls, we incubated fungi and bluegrass samples with substrate-free quantum dots as well as unbound quantum dot substrate mixtures. Penicillium fungi, annual bluegrass, and arbuscular mycorrhizal fungi all showed uptake and translocation of quantum dot-labeled organic nitrogen, but no uptake of quantum dot controls. Additionally, we observed quantum dot-labeled organic nitrogen within soil hyphae, plant roots, and plant shoots using field imaging techniques. This experiment is one of the first to demonstrate direct uptake of organic nitrogen by arbuscular mycorrhizal fungi. PMID- 19294918 TI - Rain forest nutrient cycling and productivity in response to large-scale litter manipulation. AB - Litter-induced pulses of nutrient availability could play an important role in the productivity and nutrient cycling of forested ecosystems, especially tropical forests. Tropical forests experience such pulses as a result of wet-dry seasonality and during major climatic events, such as strong El Ninos. We hypothesized that (1) an increase in the quantity and quality of litter inputs would stimulate leaf litter production, woody growth, and leaf litter nutrient cycling, and (2) the timing and magnitude of this response would be influenced by soil fertility and forest age. To test these hypotheses in a Costa Rican wet tropical forest, we established a large-scale litter manipulation experiment in two secondary forest sites and four old-growth forest sites of differing soil fertility. In replicated plots at each site, leaves and twigs (< 2 cm diameter) were removed from a 400-m2 area and added to an adjacent 100-m2 area. This transfer was the equivalent of adding 5-25 kg/ha of organic P to the forest floor. We analyzed leaf litter mass, [N] and [P], and N and P inputs for addition, removal, and control plots over a two-year period. We also evaluated basal area increment of trees in removal and addition plots. There was no response of forest productivity or nutrient cycling to litter removal; however, litter addition significantly increased leaf litter production and N and P inputs 4-5 months following litter application. Litter production increased as much as 92%, and P and N inputs as much as 85% and 156%, respectively. In contrast, litter manipulation had no significant effect on woody growth. The increase in leaf litter production and N and P inputs were significantly positively related to the total P that was applied in litter form. Neither litter treatment nor forest type influenced the temporal pattern of any of the variables measured. Thus, environmental factors such as rainfall drive temporal variability in litter and nutrient inputs, while nutrient release from decomposing litter influences the magnitude. Seasonal or annual variation in leaf litter mass, such as occurs in strong El Nino events, could positively affect leaf litter nutrient cycling and forest productivity, indicating an ability of tropical trees to rapidly respond to increased nutrient availability. PMID- 19294919 TI - Temperature oscillation coupled with fungal community shifts can modulate warming effects on litter decomposition. AB - Diel temperature oscillations are a nearly ubiquitous phenomenon, with amplitudes predicted to change along with mean temperatures under global-warming scenarios. Impact assessments of global warming have largely disregarded diel temperature oscillations, even though key processes in ecosystems, such as decomposition, may be affected. We tested the effect of a 5 degrees C temperature increase with and without diel oscillations on litter decomposition by fungal communities in stream microcosms. Five temperature regimes with identical thermal sums (degree days) were applied: constant 3 degrees and 8 degrees C; diel temperature oscillations of 5 degrees C around each mean; and oscillations of 9 degrees C around 8 degrees C. Temperature oscillations around 8 degrees C (warming scenario), but not 3 degrees C (ambient scenario), accelerated decomposition by 18% (5 degrees C oscillations) and 31% (9 degrees C oscillations), respectively, compared to the constant temperature regime at 8 degrees C. Community structure was not affected by oscillating temperatures, although the rise in mean temperature from 3 degrees to 8 degrees C consistently shifted the relative abundance of species. A simple model using temperature-growth responses of the dominant fungal decomposers accurately described the experimentally observed pattern, indicating that the effect of temperature oscillations on decomposition in our warming scenario was caused by strong curvilinear responses of species to warming at low temperature, particularly of the species becoming most abundant at 8 degrees C (Tetracladium marchalianum). These findings underscore the need to consider species-specific temperature characteristics in concert with changes in communities when assessing consequences of global warming on ecosystem processes. PMID- 19294920 TI - Long-term disease dynamics in lakes: causes and consequences of chytrid infections in Daphnia populations. AB - Understanding the drivers and consequences of disease epidemics is an important frontier in ecology. However, long-term data on hosts, their parasites, and the corresponding environmental conditions necessary to explore these interactions are often unavailable. We examined the dynamics of Daphnia pulicaria, a keystone zooplankter in lake ecosystems, to explore the long-term causes and consequences of infection by a chytridiomycete parasitoid (Polycaryum laeve). After quantifying host-pathogen dynamics from vouchered samples collected over 15 years, we used autoregressive models to evaluate (1) hypothesized drivers of infection, including host density, water temperature, dissolved oxygen, host-food availability, and lake mixing; and (2) the effects of epidemics on host populations. Infection was present in most years but varied widely in prevalence, from < 1% to 34%, with seasonal peaks in early spring and late fall. Within years, lake stratification strongly inhibited P. laeve transmission, such that epidemics occurred primarily during periods of water mixing. Development of the thermocline likely reduced transmission by spatially separating susceptible hosts from infectious zoospores. Among years, ice duration and cumulative snowfall correlated negatively with infection prevalence, likely because of reductions in spring phytoplankton and D. pulicaria density in years with extended winters. Epidemics also influenced dynamics of the host population. Infected D. pulicaria rarely (< 1%) contained eggs, and P. laeve prevalence was positively correlated with sexual reproduction in D. pulicaria. Analyses of D. pulicaria density dependent population dynamics predicted that, in the absence of P. laeve infection, host abundance would be 11-50% higher than what was observed. By underscoring the importance of complex physical processes in controlling host parasite interactions and of epidemic disease in influencing host populations, our results highlight the value of long-term data for understanding wildlife disease dynamics. PMID- 19294921 TI - Multiscale control of flooding and riparian-forest composition in Lower Michigan, USA. AB - Despite general agreement that river-valley hydrology shapes riparian ecosystems, relevant processes are difficult to distinguish and often inadequately specified in riparian studies. We hypothesize that physical constraints imposed by broad scale watershed characteristics and river valleys modify local site conditions in a predictable and probabilistic fashion. To test this hypothesis, we employ a series of structural equations that decompose occurrence of riparian ecotypes into regional temperature, catchment storm response, valley hydraulics, and local site wetness via a priori specification of factor structure and ask (1) Is there evidence for multiscale hydrologic control of riparian diversity across Lower Michigan? (2) Do representations of key constraints on flood dynamics distinguish regional patterns of riparian vegetation? (3) How important are these effects? Cross-correlation among geospatial predictors initially obscured much of the variation revealed through analysis of semipartial variance. Causal relationships implied by our model fit with observed variation in riparian conditions (chi square P = 0.43) and accounted for between 84% and 99% of the occurrence probability of five riparian ecotypes at 94 locations. Results suggest strong variation in the effects of regional climate, and both the relative importance and spatial scale of hydrologic factors influencing riparian vegetation through explicit quantification of relative flood frequency, duration, intensity, and relative overall inundation. Although climate and hydrology are not the only determinants of riparian conditions, interactions of hydrologic sourcing and flood dynamics described by our spatial models drive a significant portion of the variation in riparian ecosystem character throughout Lower Michigan, USA. PMID- 19294922 TI - No increase in colonization rate of boreal bryophytes close to propagule sources. AB - Knowledge of the process of recolonization, and which temporal and spatial scale it operates on, is central to our understanding of species distributions, metapopulation dynamics, regional extinction risks, and ecosystem resilience. In this study the recolonization pattern of boreal forest bryophytes was investigated in stands that had been clear-cut approximately 50 years ago. Species known to be sensitive to clear-cutting were inventoried in 23 mature forest stands and in adjacent young stands at 10, 20, 40, and 80 m from the former forest-clear-cut edge. Based on previous studies showing that bryophytes tend to be dispersal limited at local population levels, it was hypothesized that the recolonizaton of many bryophyte species should be higher closer to the mature forest edge. It was also hypothesized that some species would show full recovery, while for others the young stands would still be inhospitable. All these patterns were found for individual species, but the main pattern was, however, quite different. Most species had started to recolonize the young stands (i.e., little or much, depending on species), but without any tendency of a higher colonization rate close to the mature stands. Possible explanations for the limited signs of positive influence of local propagule sources might be microsite limitation or that the local propagule availability displays a rapid decline from its sources and is masked by a higher regional propagule rain. For organisms with light propagules able to build up a regional background level, the role of mature forest stands in the recolonization process of the matrix may rather be to contribute to the regional background level of spores in the landscape than to affect the adjacent stands directly. PMID- 19294923 TI - Coexistence of annual plants: generalist seed predation weakens the storage effect. AB - We investigate the effect of seed predation on the coexistence of competing annual plants. We demonstrate a role for predation that is opposite to the conventional wisdom that predation promotes coexistence by reducing the intensity of competition. In the common situation where competitive coexistence involves intraspecific competition exceeding interspecific competition, predation can undermine coexistence by reducing the overall magnitude of competition, replacing competition with "apparent competition" in a way that does not lead to differential intraspecific and interspecific effects. We demonstrate this outcome in the case where coexistence occurs by "the storage effect" in a variable environment. The storage effect arises when the environment interacts with competition to create opportunities for species to increase from low density. Critical to the storage effect is positive covariance between the response of population growth to the environment and its response to competition, when a species is at high density. This outcome prevents species at high density from taking advantage of favorable environmental conditions. A species at low density has lower covariance and can take advantage of favorable environmental conditions, giving it an advantage over a high-density species, fostering its recovery from low density. Hence, species coexistence is promoted. Here we find that density-dependent predation lowers population densities, and so weakens competition, replacing competition with apparent competition, which does not covary with the environment. As a consequence, covariance between environment and competition is weakened, reducing the advantage to a species at low density. The species still strongly interact through the combination of competition and apparent competition, but the reduced low-density advantage reduces their ability to coexist. Although this result is demonstrated specifically for the storage effect with a focus on annual plant communities, the principles involved are general ones. PMID- 19294924 TI - Latitudinal variation in herbivore pressure in Atlantic Coast salt marshes. AB - Despite long-standing interest in latitudinal variation in ecological patterns and processes, there is to date weak and conflicting evidence that herbivore pressure varies with latitude. We used three approaches to examine latitudinal variation in herbivore pressure in Atlantic Coast salt marshes, focusing on five abundant plant taxa: the grass Spartina alterniflora, the congeneric rushes Juncus gerardii and J. roemerianus, the forb Solidago sempervirens, and the shrubs Iva frutescens and Baccharis halimifolia. Herbivore counts indicated that chewing and gall-making herbivores were typically > or = 10 times more abundant at low-latitude sites than at high-latitude sites, but sucking herbivores did not show a clear pattern. For two herbivore taxa (snails and tettigoniid grasshoppers), correctly interpreting latitudinal patterns required an understanding of the feeding ecology of the species, because the species common at high latitudes did not feed heavily on plant leaves whereas the related species common at low latitudes did. Damage to plants from chewing herbivores was 2-10 times greater at low-latitude sites than at high-latitude sites. Damage to transplanted "phytometer" plants was 100 times greater for plants transplanted to low- than to high-latitude sites, and two to three times greater for plants originating from high- vs. low-latitude sites. Taken together, these results provide compelling evidence that pressure from chewing and gall-making herbivores is greater at low vs. high latitudes in Atlantic Coast salt marshes. Sucking herbivores do not show this pattern and deserve greater study. Selective pressure due to greater herbivore damage at low latitudes is likely to partially explain documented patterns of low plant palatability to chewing herbivores and greater plant defenses at low latitudes, but other factors may also play a role in mediating these geographic patterns. PMID- 19294925 TI - Habitat edges, within-patch dispersion of hosts, and parasitoid oviposition behavior. AB - Although density-edge effects are commonly reported, we have only scratched the surface in understanding the mechanisms underlying how habitat edges mediate species interactions. Here, I use a combination of field surveys and experiments to explore the linkages between the presence of host-plant edges (Scolochloa festucacea), the within-patch distribution of planthopper eggs (Delphacodes scolochloa), and the oviposition behavior of their egg parasitoids (Anagrus spp.). The field surveys revealed that densities of hosts and parasitoids at the edge were > or = 2.5 times lower than densities in the patch interior and that the effect was independent of patch size. As a consequence of its edge-avoidance behavior, host within-patch dispersion was significantly more aggregated in the presence, as compared to the absence, of an edge. Also, as patch size decreased, the proportion of the patch that was core (i.e., beyond the influence of the edge) decreased, and the degree of host aggregation increased. In a subsequent field experiment, I found that the dispersion of hosts affected Anagrus oviposition behavior only when hosts were present on islands with discrete edges. Under these circumstances, the proportion parasitized and per capita parasitized were 2.3 and 3.4 times higher, respectively, when hosts were clumped as compared to when hosts were uniformly or randomly distributed. Based on a laboratory experiment using small S. festucacea patches, I found that Anagrus had 40% shorter step lengths, spent 52% more time in the patch, and parasitized 84% more hosts when hosts were clumped as compared to uniformly distributed. These results were indicative of area-restricted search by Anagrus, which is an effective foraging strategy when hosts are clumped. This is the first study to demonstrate that predator foraging behavior in response to prey dispersion can be mediated by the presence of a patch edge. Also, because edge-averse behavior is commonly reported in the literature, an underappreciated effect of fragmentation on predator-prey interactions and stability could arise from edge-mediated effects on prey within-patch aggregation. PMID- 19294926 TI - Invasive fire ants alter behavior and morphology of native lizards. AB - Nonnative species introductions are becoming more common, but long-term consequences of the novel pressures imposed by invaders on native species remain poorly known. The red imported fire ant, Solenopsis invicta, is an invasive species with potential global impact. Comparison of lizards across the invasive range within the United States reveals novel antipredator strategies and altered morphologies that mitigate potentially lethal attack by these ants, within 70 years of their introduction. The likelihood that adult lizards will behaviorally respond to fire ant attack increases with time since invasion, but hatchlings exhibit high levels of antipredator behavior irrespective of their site of origin. Adults and hatchlings from sites invaded longer ago also have relatively longer hind limbs. This trait increases the effectiveness of behavioral strategies for removing ants and is likely an adaptive response to minimize envenomation during attack. The observed changes are not correlated with habitat, exposure to fire ants, or latitude, arguing against phenotypic plasticity and learning as causal mechanisms, and museum specimens show that morphological differences were not evident prior to fire ant invasion. These data contribute to our growing awareness that ecological invasions can prompt adaptive responses, altering the nature of interactions between invaders and the natives they contact. PMID- 19294927 TI - Age-specific reproduction and survival of individually marked Wood Thrushes, Hylocichla mustelina. AB - Several nonexclusive, sometimes contradictory, hypotheses have been employed to explain age-related changes in reproduction and survival among vertebrates. We evaluated these hypotheses with observations collected from a 31-year study of individually marked Wood Thrushes (Hylocichla mustelina). Age-specific fledgling production, survival probabilities, and derived life table columns are presented here. Additionally, we report "genetic value," or the number of young remaining to be recruited into the breeding population by individuals of age x. Controlling for age of mate, the presence of some individuals in multiple years, and the effect of year of breeding, reproduction and survival peaked in middle-aged birds of each sex and declined later in life for the oldest individuals. We could not exclude the selection hypothesis (removal of inferior individuals) as an explanation for improved female reproduction until middle age. The age at last reproduction was a significant determinant of age-related reproductive success, indicating a disappearance of low-quality individuals. Yearling females that did not successfully reproduce had greater within-season emigration than all other age classes of each sex, regardless of the reproductive success of other classes. For males, we favored the constraint hypothesis (improved reproduction with experience) to explain greater reproductive output with increasing age until at least middle age. For each sex, the age of the mate independently influenced all measures of reproductive performance, but there was generally a linear or quadratic, rather than interactive, effect of the mate's age on reproduction. For both sexes, hypotheses of age-related reproduction were only partially supported because of effects of senescence, or a decline in reproductive output or survival probabilities with increasing age. Senescence may confound attempts to examine or apply general hypotheses of age-related reproduction that assume a monotonic relationship between age and reproduction or survival. PMID- 19294928 TI - Simultaneous positive and negative density-dependent dispersal in a colonial bird species. AB - Contradictory patterns of density-dependent animal dispersal can potentially be reconciled by integrating the conspecific attraction hypothesis with the traditional competition hypothesis. We propose a hypothesis that predicts a U shaped relationship between density and both natal and breeding dispersal distance. Using 10 years of observations on a breeding colony of the Blue-footed Booby (Sula nebouxii), the hypothesis was confirmed by documenting simultaneous positive and negative density-dependent dispersal distances in natal and breeding dispersal of males and breeding dispersal of females within the colony. Point pattern analyses demonstrated that the breeding sites of Blue-footed Boobies were highly aggregated in all years within a large study area, and aggregation presumably resulted in heterogeneity in patch density throughout the colony. As predicted, at moderate to high densities, dispersal distances showed positive density dependence, with individuals moving to lower density patches. In contrast, at low to moderate densities, dispersal distances showed negative density dependence, with individuals moving to higher density patches. In both sexes of the 1994 cohort, the higher the mean density in patches used by an individual over the long term (up to age 11 years), the fewer fledglings it produced. A positive effect of density on long-term reproductive success was not detected, possibly because birds that failed during pair formation or incubation were not sampled. Density of conspecifics may be an important influence on habitat selection of breeders, and dispersal may tend to carry individuals to patches where pair formation opportunities are better and negative effects of competition on reproductive success are reduced. PMID- 19294929 TI - Quantifying a dynamic risk landscape: heterogeneous predator activity and implications for prey persistence. AB - Spatial heterogeneity in predation risk can ameliorate impacts on prey populations, particularly for prey of generalists. Spatially heterogeneous risk implies the existence of refugia, and the spatial scale of those refugia and their persistence over time affect whether prey can avoid predation by aggregating therein. Our objective was to quantify the magnitude, spatial scale, and temporal persistence of heterogeneity in risk of predation by white-footed mice (Peromyscus leucopus), an abundant generalist predator of gypsy moths (Lymantria dispar) and songbirds. We used track plates to measure white-footed mouse activity at > 170 trees in each of three forest plots in upstate New York during summers of 2003-2005. We quantified the mean and coefficient of variation of track activity among trees by fitting the beta-binomial distribution to data from each plot and study period. We measured temporal persistence by disattenuated autocorrelation, and spatial scale by fitting exponential variograms. Mice were much less abundant in 2005 than the other two years, leading to lower overall track activity but higher coefficient of variation among trees. Mouse track activity at individual trees was positively autocorrelated between monthly study periods in 2003 and 2004, and even between the two years, whereas temporal autocorrelation in 2005 was much weaker. Track activity showed positive spatial autocorrelation over lag distances from approximately 30 to > 1000 m. These findings indicate that mouse activity, and hence risk to their prey, varies substantially in space at spatial and temporal scales that appear responsive to mouse population dynamics. The spatial scale and temporal persistence of that variation imply that prey may benefit from returning to, or failing to disperse from, refugia. PMID- 19294930 TI - Hierarchical drivers of reef-fish metacommunity structure. AB - Coral reefs are highly complex ecological systems, where multiple processes interact across scales in space and time to create assemblages of exceptionally high biodiversity. Despite the increasing frequency of hierarchically structured sampling programs used in coral-reef science, little progress has been made in quantifying the relative importance of processes operating across multiple scales. The vast majority of reef studies are conducted, or at least analyzed, at a single spatial scale, ignoring the implicitly hierarchical structure of the overall system in favor of small-scale experiments or large-scale observations. Here we demonstrate how alpha (mean local number of species), beta diversity (degree of species dissimilarity among local sites), and gamma diversity (overall species richness) vary with spatial scale, and using a hierarchical, information theoretic approach, we evaluate the relative importance of site-, reef-, and atoll-level processes driving the fish metacommunity structure among 10 atolls in French Polynesia. Process-based models, representing well-established hypotheses about drivers of reef-fish community structure, were assembled into a candidate set of 12 hierarchical linear models. Variation in fish abundance, biomass, and species richness were unevenly distributed among transect, reef, and atoll levels, establishing the relative contribution of variation at these spatial scales to the structure of the metacommunity. Reef-fish biomass, species richness, and the abundance of most functional-groups corresponded primarily with transect-level habitat diversity and atoll-lagoon size, whereas detritivore and grazer abundances were largely correlated with potential covariates of larval dispersal. Our findings show that (1) within-transect and among-atoll factors primarily drive the relationship between alpha and gamma diversity in this reef fish metacommunity; (2) habitat is the primary correlate with reef-fish metacommunity structure at multiple spatial scales; and (3) inter-atoll connectedness was poorly correlated with the nonrandom clustering of reef-fish species. These results demonstrate the importance of modeling hierarchical data and processes in understanding reef-fish metacommunity structure. PMID- 19294931 TI - Elicitation by design in ecology: using expert opinion to inform priors for Bayesian statistical models. AB - Bayesian statistical modeling has several benefits within an ecological context. In particular, when observed data are limited in sample size or representativeness, then the Bayesian framework provides a mechanism to combine observed data with other "prior" information. Prior information may be obtained from earlier studies, or in their absence, from expert knowledge. This use of the Bayesian framework reflects the scientific "learning cycle," where prior or initial estimates are updated when new data become available. In this paper we outline a framework for statistical design of expert elicitation processes for quantifying such expert knowledge, in a form suitable for input as prior information into Bayesian models. We identify six key elements: determining the purpose and motivation for using prior information; specifying the relevant expert knowledge available; formulating the statistical model; designing effective and efficient numerical encoding; managing uncertainty; and designing a practical elicitation protocol. We demonstrate this framework applies to a variety of situations, with two examples from the ecological literature and three from our experience. Analysis of these examples reveals several recurring important issues affecting practical design of elicitation in ecological problems. PMID- 19294932 TI - Major dimensions in food-web structure properties. AB - The covariance among a range of 20 network structural properties of food webs plus net primary productivity was assessed for 14 published food webs using principal components analysis. Three primary components explained 84% of the variability in the data sets, suggesting substantial covariance among the properties employed in the literature. The first dimension explained 48% of the variance and could be ascribed to connectance, covarying significantly with the proportion of intermediate species and characteristic path length. The second dimension explained 19% and was related to trophic species richness. The third axis explained 17% and was related to ecosystem net primary productivity. A distinct opposite clustering of connectance, the proportion of intermediate species, and mean trophic level vs. the proportion of top and basal species and path length suggests a dichotomy in food-web structure. Food webs appear either clustered and highly interconnected or elongated with fewer links. PMID- 19294933 TI - [Profit through safety: safety plan for a hospital in need]. PMID- 19294934 TI - [One opinion is not the next: quantitative analysis of IGZ inspectors variation ]. PMID- 19294935 TI - [Patient safety: unintentional and potentially preventive damage by cutting specialists]. PMID- 19294936 TI - [Multidisciplinary care of patient's safety at the child intensive care]. PMID- 19294937 TI - ["The dear friend syndrome": if the patient is a colleague and the colleague is the doctor]. PMID- 19294938 TI - [Therapy resistance hypertension: always measure for 24 hours]. PMID- 19294939 TI - [Responsible care: the lessons learned from recent incidences]. PMID- 19294940 TI - [Radiation exposure in computed tomography in the Netherlands: risk-benefit analysis]. PMID- 19294941 TI - [A women with a strange shape of her knee prosthesis]. PMID- 19294942 TI - [Unintentional overdose by third party: patient controlled analgesia by a child after operation]. PMID- 19294943 TI - [Isopropanol toxicity]. PMID- 19294944 TI - [The discussion of medical errors with the patient]. PMID- 19294945 TI - [Jurisprudence judging mistakes made during telephone triage]. PMID- 19294946 TI - [Resign!]. PMID- 19294947 TI - Sleep--an affair of the heart. PMID- 19294948 TI - Still missing some significant ingredients. PMID- 19294949 TI - Association of sleep duration with mortality from cardiovascular disease and other causes for Japanese men and women: the JACC study. AB - STUDY OBJECTIVES: To examine sex-specific associations between sleep duration and mortality from cardiovascular disease and other causes. DESIGN: Cohort study. SETTING: Community-based study. PARTICIPANTS: A total of 98,634 subjects (41,489 men and 57,145 women) aged 40 to 79 years from 1988 to 1990 and were followed until 2003. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: During a median follow up of 14.3 years, there were 1964 deaths (men and women: 1038 and 926) from stroke, 881 (508 and 373) from coronary heart disease, 4287 (2297 and 1990) from cardiovascular disease, 5465 (3432 and 2033) from cancer, and 14,540 (8548 and 5992) from all causes. Compared with a sleep duration of 7 hours, sleep duration of 4 hours or less was associated with increased mortality from coronary heart disease for women and noncardiovascular disease/noncancer and all causes in both sexes. The respective multivariable hazard ratios were 2.32 (1.19-4.50) for coronary heart disease in women, 1.49 (1.02-2.18) and 1.47 (1.01-2.15) for noncardiovascular disease/noncancer, and 1.29 (1.02-1.64) and 1.28 (1.03-1.60) for all causes in men and women, respectively. Long sleep duration of 10 hours or longer was associated with 1.5- to 2-fold increased mortality from total and ischemic stroke, total cardiovascular disease, noncardiovascular disease/noncancer, and all causes for men and women, compared with 7 hours of sleep in both sexes. There was no association between sleep duration and cancer mortality in either sex. CONCLUSIONS: Both short and long sleep duration were associated with increased mortality from cardiovascular disease, noncardiovascular disease/noncancer, and all causes for both sexes, yielding a U shaped relationship with total mortality with a nadir at 7 hours of sleep. PMID- 19294950 TI - Slow wave sleep and REM sleep awakenings do not affect sleep dependent memory consolidation. AB - STUDY OBJECTIVES: The effects of REM sleep and slow wave sleep (SWS) deprivation on sleep-dependent motor and declarative memory consolidation. DESIGN: Randomized, within-subject, cross-over study. SETTING: Weekly (women: monthly) sleep laboratory visits, with retest 60 hours later. PARTICIPANTS: Twelve healthy subjects (6 men) aged between 20 and 30 years. INTERVENTIONS: REM sleep deprivation, SWS deprivation, or undisturbed sleep. MEASUREMENTS AND RESULTS: We deprived subjects once each of REM sleep and SWS, and once let them sleep undisturbed through the night. After each night, we tested declarative and procedural memory consolidation. We tested memory performance by a verbal paired associate task and a sequential finger-tapping task at 21:00 on the study night and again 60 hours later. Although REM sleep and SWS awakenings led to a significant reduction of the respective sleep stages, memory consolidation remained unaffected. We also found a significant correlation between the declarative task and sleep spindles in the undisturbed condition, especially the sleep spindles in the first third of the night. CONCLUSION: We suggest that word pair learning relies on stage 2 sleep spindles and requires little SWS. Their sleep dependent consolidation is not affected by SWS deprivation. Simple motor tasks may either be consolidated in stage 2 sleep or depend on only small amounts of REM sleep. Their sleep dependent consolidation is not influenced by REM sleep deprivation. PMID- 19294951 TI - Banking sleep: realization of benefits during subsequent sleep restriction and recovery. AB - OBJECTIVE: Determine whether sleep extension (a) improves alertness and performance during subsequent sleep restriction and (b) impacts the rate at which alertness and performance are restored by post-restriction recovery sleep. DESIGN: Participants were randomly assigned to an Extended (10 h time in bed [TIB]) or Habitual TIB [mean (SD) hours = 7.09 (0.7)] sleep group for one week, followed by 1 Baseline (10 hours or habitual TIB), 7 Sleep Restriction (3 h TIB), and 5 Recovery Sleep nights (8 h TIB). Performance and alertness tests were administered hourly between 08:00-18:00 during all in-laboratory phases of the study. SETTING: Residential sleep/performance testing facility. PARTICIPANTS: Twenty-four healthy adults (ages 18-39) participated in the study. INTERVENTIONS: Extended vs. habitual sleep durations prior to sleep restriction. RESULTS: Psychomotor vigilance task (PVT) lapses were more frequent and modified maintenance of wakefulness (MWT) sleep latency was shorter in the Habitual group than in the Extended group across the sleep restriction phase. During the Recovery phase, PVT speed rebounded faster (and PVT lapsing recovered significantly after the first night of recovery sleep) in the Extended group. No group differences in subjective sleepiness were evident during any phase of the study. CONCLUSION: The extent to which sleep restriction impairs objectively measured alertness and performance, and the rate at which these impairments are subsequently reversed by recovery sleep, varies as a function of the amount of nightly sleep obtained prior to the sleep restriction period. This suggests that the physiological mechanism(s) underlying chronic sleep debt undergo long-term (days/weeks) accommodative/adaptive changes. PMID- 19294952 TI - Individual differences in childhood sleep problems predict later cognitive executive control. AB - STUDY OBJECTIVE: To determine whether individual differences in developmental patterns of general sleep problems are associated with 3 executive function abilities-inhibiting, updating working memory, and task shifting-in late adolescence. PARTICIPANTS: 916 twins (465 female, 451 male) and parents from the Colorado Longitudinal Twin Study. MEASUREMENTS AND RESULTS: Parents reported their children's sleep problems at ages 4 years, 5 y, 7 y, and 9-16 y based on a 7-item scale from the Child-Behavior Checklist; a subset of children (n = 568) completed laboratory assessments of executive functions at age 17. Latent variable growth curve analyses were used to model individual differences in longitudinal trajectories of childhood sleep problems. Sleep problems declined over time, with approximately 70% of children having > or = 1 problem at age 4 and approximately 33% of children at age 16. However, significant individual differences in both the initial levels of problems (intercept) and changes across time (slope) were observed. When executive function latent variables were added to the model, the intercept did not significantly correlate with the later executive function latent variables; however, the slope variable significantly (P < 0.05) negatively correlated with inhibiting (r = -0.27) and updating (r = 0.21), but not shifting (r = -0.10) abilities. Further analyses suggested that the slope variable predicted the variance common to the 3 executive functions (r = -0.29). CONCLUSIONS: Early levels of sleep problems do not seem to have appreciable implications for later executive functioning. However, individuals whose sleep problems decrease more across time show better general executive control in late adolescence. PMID- 19294953 TI - Evaluation of a school-based intervention for adolescent sleep problems. AB - STUDY OBJECTIVES: The present study investigated the effectiveness of a school based intervention in increasing sleep knowledge and improving adolescent sleep problems. DESIGN: A randomized, controlled trial using 2 groups (program class, classes-as-usual: [CAU]) assessed over 3 time points (pre-program, post-program, 6-week follow-up). PARTICIPANTS/SETTING: Eighty-one students (mean age = 15.6 +/- 0.6 y; 33% male) from 2 schools in South Australia. Schools provided one class to participate in the sleep intervention program (N = 41) and a second class to act as a control class (N = 40). INTERVENTION: Four 50-minute classes across a 4-week period. Classes consisted of educating adolescents on promoting and maintaining a healthy lifestyle based on a cognitive-behavior therapy framework. MEASUREMENTS AND RESULTS: Data were collected pre-program, post-program, and at 6-week follow up using an online questionnaire. Qualitative student and teacher data were collected at post-program. Baseline data indicated sleep problems were prevalent (53.1% insufficient sleep on school nights [< 8 h] and 77.8% discrepant school/weekend rise times [> 2 h]). These 2 criteria identified 36 adolescents with a delayed sleep timing (DST; Program, N = 21; CAU, N = 15). The program increased sleep knowledge (P = 0.001); however, analyses revealed no significant effects on target sleep variables as compared with the CAU class for the entire group (all P > 0.05). For DST adolescents, there was a significant interaction for reducing the discrepancy between school and weekend out of bed times (P = 0.002). There was no impact on other sleep parameters or depressed mood. CONCLUSIONS: School-based sleep interventions for adolescents are a novel method for addressing a prevalent problem. Future programs should develop ways to motivate adolescents to change sleep practices. PMID- 19294954 TI - Sleep disturbances in prepubertal children with attention deficit hyperactivity disorder: a home polysomnography study. AB - STUDY OBJECTIVE: To examine sleep architecture and reported sleep problems in children with ADHD and normal controls, while considering the roles of pertinent moderating factors. DESIGN: Overnight sleep recordings were conducted in 15 children diagnosed with ADHD (DSM-IV) without comorbid psychiatric problems and in 23 healthy controls aged 7 to 11 years. Children were on no medication, in good health and did not consume products containing caffeine > or = 7 days prior to the polysomnography (PSG) study. PSG evaluation was performed at each child's home; children slept in their regular beds and went to bed at their habitual bedtimes. MEASUREMENTS: Standard overnight multichannel PSG evaluation was performed using a portable polysomnography device. In addition, parents were asked to complete a sleep questionnaire. RESULTS: Compared to controls, children in the ADHD group had significantly shorter duration of REM sleep, smaller percentage of total sleep time spent in REM sleep, and shorter sleep duration. In addition, the ADHD group had higher scores on the insufficient sleep and sleep anxiety factors than children in the control group. CONCLUSION: The present findings support the hypothesis that children with ADHD present sleep disturbances. PMID- 19294955 TI - Efficacy and safety of 6-month nightly ramelteon administration in adults with chronic primary insomnia. AB - STUDY OBJECTIVES: Long-duration (> or = 6 months) polysomnographic studies of insomnia medications are lacking. This study evaluated the long-term efficacy of ramelteon, a selective MT1/MT2 melatonin-receptor agonist used for insomnia treatment. DESIGN: Six-month, randomized, double-blind, placebo-controlled study. SETTING: Forty-six investigative sites in the United States, Europe, Russia, and Australia. PARTICIPANTS: Four hundred fifty-one adults (age > or = 18 years) with chronic primary insomnia. INTERVENTIONS: Ramelteon, 8 mg, or placebo 30 minutes before bedtime nightly for 6 months. MEASUREMENTS: Sleep was evaluated by polysomnography and morning questionnaires on the first 2 nights of Week 1; the last 2 nights of Months 1, 3, 5, and 6; and Nights 1 and 2 of the placebo run out. Next-morning residual effects as well as adverse effects and vital signs were recorded at each visit. Rebound insomnia and withdrawal effects were evaluated during placebo run-out. RESULTS: Over the 6 months of treatment, ramelteon consistently reduced latency to persistent sleep compared with baseline and with placebo; significant decreases were observed at Week 1 and Months 1, 3, 5, and 6 (P < 0.05). Ramelteon significantly reduced subjective sleep latency relative to placebo at Week 1, Month 1, and Month 5 (P < 0.05), with reductions nearing statistical significance at Months 3 and 6 (P < or = 0.08). No significant next-morning residual effects were detected during ramelteon treatment. No withdrawal symptoms or rebound insomnia were detected after ramelteon discontinuation. Most adverse events were mild or moderate in severity. CONCLUSIONS: In adults with chronic insomnia, long-term ramelteon treatment consistently reduced sleep onset, with no next-morning residual effects or rebound insomnia or withdrawal symptoms upon discontinuation. PMID- 19294956 TI - Airway dilator muscle activity and lung volume during stable breathing in obstructive sleep apnea. AB - STUDY OBJECTIVES: Many patients with obstructive sleep apnea (OSA) have spontaneous periods of stable flow limited breathing during sleep without respiratory events or arousals. In addition, OSA is often more severe during REM than NREM and more severe during stage 2 than slow wave sleep (SWS). The physiological mechanisms for these observations are unknown. Thus we aimed to determine whether the activity of two upper airway dilator muscles (genioglossus and tensor palatini) or end-expiratory lung volume (EELV) differ between (1) spontaneously occurring stable and cyclical breathing and (2) different sleep stages in OSA. DESIGN: Physiologic observation. SETTING: Sleep physiology laboratory. STUDY PARTICIPANTS: 15 OSA patients with documented periods of spontaneous stable breathing. INTERVENTION: Subjects were instrumented with intramuscular electrodes for genioglossus and tensor palatini electromyograms (EMG(GG) and EMG(TP)), chest and abdominal magnetometers (EELV measurement), an epiglottic pressure catheter (respiratory effort), and a mask and pneumotachograph (airflow/ventilation). Patients slept supine overnight without CPAP. MEASUREMENTS AND RESULTS: Peak and Tonic EMG(GG) were significantly lower during cyclical (85.4 +/- 2.7 and 94.6 +/- 4.7 % total activity) than stable breathing (109.4 +/- 0.4 and 103 +/- 0.8% total activity, respectively). During respiratory events in REM, tonic EMG(GG) activity was lower than during respiratory events in stage 2 (71.9 +/- 5.1 and 119.6 +/- 5.6% total activity). EMG(GG) did not differ between stable stage 2 and stable SWS (98.9 +/- 3.2 versus 109.7 +/- 4.4% total activity), nor did EMG(TP) or EELV differ in any breathing condition/sleep stage. CONCLUSIONS: Increased genioglossus muscle tone is associated with spontaneous periods of stable flow limited breathing in the OSA subjects studied. Reductions in genioglossus activity during REM may explain the higher severity of OSA in that stage. Increased lung volume and tensor palatini activity do not appear to be major mechanisms enabling spontaneous stable flow limited breathing periods. PMID- 19294957 TI - Relationship between obstructive sleep apnea severity and brain activation during a sustained attention task. AB - STUDY OBJECTIVES: The objectives of this study were to (1) characterize cognitive and cerebral correlates of attention and response speed in patients with obstructive sleep apnea (OSA) and (2) assess the association of performance and brain activation with measures of OSA severity. DESIGN: Patients with OSA and controls were compared on performance and brain activation during a sustained attention task. The association of reaction time and brain activation with apnea hypopnea index, nocturnal hypoxia, and arousals was assessed. SETTING: Functional magnetic resonance imaging was conducted while participants performed a Go-No-Go task. The 'Go' trials of the Go-No-Go task were used to index attention processing. PARTICIPANTS: Fourteen patients with OSA and 14 normal control subjects with equivalent age, body mass index, blood pressure, and education. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Patients with OSA showed decreased brain activation in cingulate, frontal, and parietal regions typically involved in attention tasks, compared with control subjects. Within the patients with OSA, increasing arousal index, but not desaturation index, was associated with slower mean reaction time and with decreased brain activation in areas involved in arousal and attention, response selection, motor response, and decision making. The apnea-hypopnea index, by itself, was not associated with changes in cerebral response. CONCLUSIONS: Patients with OSA showed decreased brain activation compared with control subjects during an attention task. The association of arousal index (but not hypoxia) with slow reaction times and brain activation suggests that alertness and reaction times show greater correlations with measures of sleep disruption than with measures of hypoxia. PMID- 19294958 TI - Daytime sleepiness and driving performance in patients with obstructive sleep apnea: comparison of the MSLT, the MWT, and a simulated driving task. AB - STUDY OBJECTIVES: To test the reliability of a driving-simulation test for the objective measurement of daytime alertness compared with the Multiple Sleep Latency Test (MSLT) and with the Maintenance of Wakefulness Test (MWT), and to test the ability to drive safely, in comparison with on-road history, in the clinical setting of untreated severe obstructive sleep apnea. DESIGN: N/A. SETTING: Sleep laboratory. PATIENTS OR PARTICIPANTS: Twenty-four patients with severe obstructive sleep apnea and reported daytime sleepiness varying in severity (as measured by the Epworth Sleepiness Scale). INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Patients underwent MSLT and MWT coupled with 4 sessions of driving-simulation test on 2 different days randomly distributed 1 week apart. Simulated-driving performance (in terms of lane-position variability and crash occurrence) was correlated with sleep latency on the MSLT and more significantly on the MWT, showing a predictive validity toward the detection of sleepy versus alert patients with obstructive sleep apnea. In addition, patients reporting excessive daytime sleepiness or a history of car crashes showed poorer performances on the driving simulator. CONCLUSIONS: A simulated driving test is a suitable tool for objective measurement of daytime alertness in patients with obstructive sleep apnea. Further studies are needed to clarify the association between simulated-driving performance and on-road crash risk of patients with sleep disordered breathing. PMID- 19294959 TI - Modeling napping, post-lunch dip, and other variations in human sleep propensity. AB - STUDY OBJECTIVES: To model sleep propensity (SP) as a continuous variable across 24 hours and to model the post-noon nap zone, or post-lunch dip in performance, and the early evening trough in SP. METHODS: The present model is a variant of the 2-process model with 2 major modifications. (1) The circadian threshold process was replaced by sleep drive R, derived from REM sleep propensity, which shows a strong circadian modulation. (2) The model is based on a multiplicative interaction between the 2 input variables S and R. The model parameters S and R were estimated from experimental data. Thus, SP is modeled by multiplicative interaction of 2 sleep drives, S and R, the former of homeostatic, the latter of circadian nature. In short: SP = S x R. RESULTS: Under the condition of normal phase and duration of nighttime sleep, SP across 24 hours displays 4 characteristics, (a) a major peak at nighttime, (b) a secondary increase, which peaks post-noon, (c) a first local minimum at sleep offset in the morning, and (d) a second local minimum in the early evening hours. Model simulations with either delayed or advanced sleep times suggest that the magnitude of the post noon nap zone depends on the phase of the major sleep period within 24 hours. While the nap zone is attenuated or disappears when night sleep is delayed, SP increases during daytime when night sleep is advanced. In all conditions, the evening local minimum of SP remained stable. CONCLUSIONS: SP can be modeled as a continuous variable, based on the multiplicative interaction of 2 basic sleep drives. The model predictions are in agreement with known variations of SP across 24 hours. PMID- 19294960 TI - Auditory inhibition of rapid eye movements and dream recall from REM sleep. AB - STUDY OBJECTIVES: There is debate in dream research as to whether ponto-geniculo occipital (PGO) waves or cortical arousal during sleep underlie the biological mechanisms of dreaming. This study comprised 2 experiments. As eye movements (EMs) are currently considered the best noninvasive indicator of PGO burst activity in humans, the aim of the first experiment was to investigate the effect of low-intensity repeated auditory stimulation on EMs (and inferred PGO burst activity) during REM sleep. It was predicted that such auditory stimuli during REM sleep would have a suppressive effect on EMs. The aim of the second experiment was to examine the effects of this auditory stimulation on subsequent dream reporting on awakening. DESIGN: Repeated measures design with counterbalanced order of experimental and control conditions across participants. SETTING: Sleep laboratory based polysomnography (PSG) PARTICIPANTS: Experiment 1 : 5 males and 10 females aged 18-35 years (M = 20.8, SD = 5.4). Experiment 2 : 7 males and 13 females aged 18-35 years (M = 23.3, SD = 5.5). INTERVENTIONS: Below waking threshold tone presentations during REM sleep compared to control REM sleep conditions without tone presentations. MEASUREMENTS AND RESULTS: PSG records were manually scored for sleep stages, EEG arousals, and EMs. Auditory stimulation during REM sleep was related to: (a) an increase in EEG arousal, (b) a decrease in the amplitude and frequency of EMs, and (c) a decrease in the frequency of visual imagery reports on awakening. CONCLUSIONS: The results of this study provide phenomenological support for PGO-based theories of dream reporting on awakening from sleep in humans. PMID- 19294961 TI - Evidence of dissociated arousal states during NREM parasomnia from an intracerebral neurophysiological study. AB - STUDY OBJECTIVES: Arousal parasomnias are expressions of sleep/ wake state dissociations in which wakefulness and NREM sleep seem to coexist. We describe the results of a neurophysiological (intracerebral EEG) investigation that captured an episode of confusional arousal. DESIGN: Observational analysis. SETTING: Tertiary sleep center. SUBJECT: A 20-year-old male with refractory focal epilepsy. MEASUREMENTS AND RESULTS: The intracerebral EEG findings documented the presence of a local arousal of the motor and cingulate cortices associated with increased delta activity in the frontoparietal associative cortices; these findings were noted preceding the onset and persisting throughout the episode. CONCLUSIONS: The presence of dissociated sleep/wake states in confusional arousals is the expression not of a global phenomenon, but rather of the coexistence of different local states of being: arousal of the motor and cingulate cortices and inhibition of the associative ones. Whether this is an exclusive feature of NREM parasomnias, or a common substrate on which other triggering elements act, needs to be clarified. PMID- 19294962 TI - Fermented ginseng improves the first-night effect in humans. AB - STUDY OBJECTIVES: The goal of this study was to clarify whether ginseng fermented by lactic acid bacteria (fermented ginseng, FG), can improve the first-night effect (FNE) in humans. DESIGN: Behavioral tests and quantification of mRNA expression related to GABAergic neurotransmission in brain (glutamic acid decarboxylase 1, gamma-aminobutyrate aminotransferase [Abat], gamma-aminobutyric acid transporter 1 [GAT1], gamma-aminobutyric acid transporter 4, gamma aminobutyric acid A receptor subunit alpha 1 and gamma-aminobutyric acid A receptor subunit alpha 2) were carried out in FG-treated mice. We also performed double-blind sleep recordings of human subjects given FG or placebo. SETTING: A university-based sleep laboratory. PATIENTS OR PARTICIPANTS: Sixteen healthy male volunteers (aged 20.69 +/- 0.44 years) were observed in the human study. INTERVENTIONS: At the end of administration, 2 consecutive all-night polysomnography recordings were performed. Subjects also completed psychological questionnaires, and urine and saliva samples were taken to analyze stress sensitive markers. MEASUREMENTS AND RESULTS: The light-dark transition test demonstrated that FG had some anxiolytic effect in mice, but other anxiety measures were unaffected. The hippocampal mRNA expression showed a decrease of Abat and GAT1 suggesting an increase of GABA. Other regions (amygdala and cerebellum) showed no differences. Furthermore, there was some evidence (using simple pairwise comparisons but not supported in the full ANOVA model) that administration of FG tended to diminish decreases in total sleep time and sleep efficiency (seen as first night effects in the placebo group) without affecting sleep architecture. CONCLUSIONS: Our results suggest the administration of FG could improve the FNE in humans. The improvement may be related to an anxiolytic effect of FG which acts via GABAergic modification. PMID- 19294963 TI - Season of birth, gender, and social-cultural effects on sleep timing preferences in humans. AB - STUDY OBJECTIVE: The present study aimed to analyze season of birth effects on preferred sleep-wake cycle timing as assessed by Morningness-Eveningness Questionnaire (MEQ). PARTICIPANTS AND MEASUREMENTS: The MEQ was administered to a sample of 5,720 university students (3,851 Italians and 1,869 Spaniards; 3,877 female and 1,843 male; mean age 22.23 +/- 2.98 years). RESULTS: Females preferred to go to bed significantly earlier and sleep longer than males, regardless of season of birth and nationality. Subjects born in spring and summer went to bed and reached midpoint of sleep later than subjects born in fall and winter. Nationality significantly affected all the sleep parameters considered except duration. CONCLUSION: Overall, the effect of the season of birth on sleep preference timing was significant but quantitatively small. We suggest an evolutionary context for the different contributions of genetic and environmental factors in modulating sleep-wake cycles in humans. PMID- 19294964 TI - Ambulatory surgery in the United States, 2006. AB - OBJECTIVES: This report presents national estimates of surgical and nonsurgical procedures performed on an ambulatory basis in hospitals and freestanding ambulatory surgery centers in the United States during 2006. Data are presented by types of facilities, age and sex of the patients, and geographic regions. Major categories of procedures and diagnoses are shown by age and sex. Selected estimates are compared between 1996 and 2006. METHODS: The estimates are based on data collected through the 2006 National Survey of Ambulatory Surgery by the Centers for Disease Control and Prevention's National Center for Health Statistics (NCHS). The survey was conducted from 1994-1996 and again in 2006. Diagnoses and procedures presented are coded using the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM). RESULTS: In 2006, an estimated 57.1 million surgical and nonsurgical procedures were performed during 34.7 million ambulatory surgery visits. Of the 34.7 million visits, 19.9 million occurred in hospitals and 14.9 million occurred in freestanding ambulatory surgery centers. The rate of visits to freestanding ambulatory surgery centers increased about 300 percent from 1996 to 2006, whereas the rate of visits to hospital-based surgery centers remained largely unchanged during that time period. Females had significantly more ambulatory surgery visits (20.0 million) than males (14.7 million), and a significantly higher rate of visits (132.0 per 1000 population) compared with males (100.4 per 1000 population). Average times for surgical visits were higher for ambulatory surgery visits to hospital-based ambulatory surgery centers than for visits to freestanding ambulatory surgery centers for the amount of time spent in the operating room (61.7 minutes compared with 43.2 minutes), the amount of time spent in surgery (34.2 minutes compared with 25.1 minutes), the amount of time spent in the postoperative recovery room (79.0 minutes compared with 53.1 minutes), and overall time (146.6 minutes compared with 97.7 minutes). Although the majority of visits had only one or two procedures performed (56.3 percent and 28.5 percent, respectively), 2.6 percent had five or more procedures performed. Frequently performed procedures on ambulatory surgery patients included endoscopy of large intestine (5.8 million), endoscopy of small intestine (3.5 million), extraction of lens (3.1 million), injection of agent into spinal canal (2.7 million), and insertion of prosthetic lens (2.6 million). The leading diagnoses at ambulatory surgery visits included cataract (3.0 million); benign neoplasms (2.0 million), malignant neoplasms (1.2 million), diseases of the esophagus (1.1 million), and diverticula of the intestine (1.1 million). PMID- 19294965 TI - Fetal and perinatal mortality, United States, 2005. AB - OBJECTIVES: This report presents 2005 fetal and perinatal mortality data by a variety of characteristics, including maternal age, marital status, race, Hispanic origin, and state of residence; and by fetal gestational age at delivery, birthweight, plurality, and sex. Trends in fetal and perinatal mortality are also examined. METHODS: Descriptive tabulations of data are presented and interpreted. RESULTS: In 2005, there were 25,894 reported fetal deaths of 20 weeks of gestation or more in the United States. The U.S. fetal mortality rate was 6.22 fetal deaths of 20 weeks of gestation or more per 1000 live births and fetal deaths, not significantly different from the rate of 6.20 in 2004 or 6.23 in 2003. The fetal mortality rate declined slowly but steadily from 1990 to 2003, but did not decline from 2003 to 2005. Most of the decline in the overall fetal mortality rate from 1990 to 2003 was due to a decline in fetal deaths of 28 weeks of gestation or more; fetal deaths of 20-27 weeks did not decline. From 2003 to 2005, fetal mortality rates did not decline for either gestational age grouping. From 2003 to 2005, fetal mortality rates declined significantly for non-Hispanic white and non-Hispanic black women, but not for Hispanic, American Indian or Alaska Native (AIAN), or Asian or Pacific Islander women. In 2005, the fetal mortality rate for non-Hispanic black women (11.13) was 2.3 times the rate for non-Hispanic white women (4.79). The rate for AIAN women (6.17) was 29% higher, and the rate for Hispanic women (5.44) was 14% higher than the rate for non-Hispanic white women. Fetal mortality rates are elevated for a number of groups, including teenagers, women aged 35 years and over, unmarried women, and multiple deliveries. In 2005, one-half of fetal deaths of 20 weeks of gestation or more occurred at 20-27 weeks of gestation. PMID- 19294966 TI - Don't overlook parish nurses. PMID- 19294968 TI - Risk management. Theft happens. Protecting the data is what matters. PMID- 19294967 TI - Health reform. Proposed federal health board could take decision-making away from Congress. PMID- 19294969 TI - Environment. Hybrid fleet fuels a bevy of benefits for hospital and home health staff. PMID- 19294970 TI - Other voices. Should money drive everything? PMID- 19294971 TI - Litigation. GPOs sue manufacturer for 'monopolistic' deals. PMID- 19294972 TI - The extra mile. A dramatic homecoming. PMID- 19294973 TI - Medical education. Hospitals, universities must nurture their partnerships. PMID- 19294974 TI - Nursing. Hospital, college link up to strengthen nurse staffing and training. PMID- 19294975 TI - Calling an IT time-out. Tough economy may present an opportunity. AB - In these tough economic times, executives are scrutinizing IT expenditures. This presents an opportunity to make sure that money is being spent wisely. PMID- 19294976 TI - Will we make history? AB - From FDR's call for a social insurance system that would extend "from cradle to grave" to George W. Bush's Medicare prescription drug plan, successive American presidents have tried, each in his own way, to improve the nation's health care system. Now it's Barack Obama's turn, and his multifaceted proposal, backed by billions of dollars in federal funding and support from a broad spectrum of stakeholders, could make comprehensive health care reform a reality. PMID- 19294977 TI - Hospitals are working to control undiagnosed diabetes. AB - Patients with high blood sugar levels are at risk for a wide range of conditions. Early testing and new treatments help keep them out of danger. PMID- 19294978 TI - Not your father's COO. AB - The lines on the organizational chart used to be pretty clear. CEOs were responsible for strategy, external issues and dealing with the board, COOs rode herd on internal operations and day-to-day management, and CFOs managed the money to make it all happen. But as with so many other areas of health care, those roles have blurred. While the chief operating officer's job description hasn't been dramatically rewritten, the pressures facing hospitals today--physician relations: an increased focus on quality, safety and patient satisfaction; reimbursement changes such as Medicare's policies concerning payment for so called never events; and a continually evolving competitive environment--are radically changing the demands on hospital COOs. The four COOs profiled here have embraced their expanded roles. Whether helping create and implement a rural organization's strategic plan, translating corporate policy to the front-line troops, leading a big regional system in a time of market crisis, or balancing the goals of the parent system with those of dozens of local hospitals, these COOs are performing jobs their predecessors a generation ago might not recognize. PMID- 19294979 TI - Something new for mental health services: hope. PMID- 19294980 TI - Seeing the light with CPOE. AB - Computerized order entry may be the right tool to improve quality, safety and efficiency, but implementing it can be scary. These hospitals explain how they did it. PMID- 19294981 TI - Inpatient service trends. PMID- 19294982 TI - Hospitals in pursuit of excellence. PMID- 19294983 TI - Molecular characterization of SAT-2 foot-and-mouth disease virus isolates obtained from cattle during a four-month period in 2001 in Limpopo Province, South Africa. AB - Foot-and-mouth disease (FMD) is an acute, highly contagious viral infection of domestic and wild cloven-hoofed animals. The virus is a single-stranded RNA virus that has a high rate of nucleotide mutation and amino acid substitution. In southern Africa the South African Territories (SAT) 1-3 serotypes of FMD virus are maintained by large numbers of African buffaloes (Syncerus caffer), which provide a potential source of infection for domestic livestock and wild animals. During February 2001, an outbreak of SAT-2 was recorded in cattle in the FMD control zone of South Africa, adjacent to the Kruger National Park (KNP). They had not been vaccinated against the disease since they form the buffer between the vaccination and free zones but in the face of the outbreak, they were vaccinated as part of the control measures to contain the disease. The virus was, however, isolated from some of them on several occasions up to May 2001. These isolates were characterized to determine the rate of genetic change in the main antigenic determinant, the 1 D/2A gene. Nucleotide substitutions at 12 different sites were identified of which five led to amino acid changes. Three of these occurred in known antigenic sites, viz. the GH-loop and C-terminal part of the protein, and two of these have previously been shown to be subject to positive selection. Likelihood models indicated that the ratio of non-synonymous to synonymous changes among the outbreak sequences recovered from cattle was four times higher than among comparable sequences isolated from wildlife, suggesting that the virus may be under greater selective pressure during rapid transmission events. PMID- 19294984 TI - Value of tests for evaluating udder health in dairy goats: somatic cell counts, California Milk Cell Test and electrical conductivity. AB - The value of electric conductivity (EC), California Milk Cell Test (CMCT) and somatic cell count (SCC) as diagnostic tools was investigated in dairy goats. Conductivity colour reading correlated with SCC. Milk samples with conductivity colour red had significantly higher SCC than those with conductivity colours green and orange (P < 0.001). There were moderate positive correlations between CMCT (R2 = 0.470), and conductivity score and CMCT and conductivity colour readings (R2 = 0.597). Conductivity scores were significantly (P< 0.001) higher during and after intra-mammary treatment with Cloxamast LC and conductivity colours were significantly different between treatment and control groups (P< 0.001). There was a weak positive correlation between conductivity colour and stage of lactation (R2 = 0.317) and a moderately positive correlation between conductivity score and stage of lactation (R2 = 0.523). A moderately negative correlation was shown between milk yield and conductivity score (R2 = -0.426) and between milk yield and conductivity colour (R2 = -0.433). Moderate positive correlations were present between CMCT and SCC (R2 = 0.689) and between CMCT and stage of lactation (R2 = 0.459). CMCT ratings were significantly different (P < 0.001) for the intra-mammary treatment groups. CMCT ratings for infected and non infected udder halves (P = 0.008) were significantly different; as were those for infected and non-infected udder halves and for left and right udder halves separately (P= 0.010). CMCT ratings for milk samples with SCC above and below 750 x 10(3) cells per ml were significantly different (P < 0.001) as well as for milk from treated and control udder halves with SCC below or above 750 x 10(3) cells per ml (P < 0.001). CMCT was found to be more accurate for indicating the absence of mastitis than for diagnosing it. There were significant differences in log SCC between treatment and control groups, during and after treatment. Infected udder halves had significantly higher log SCC than non-infected udder halves before and after treatment, but not during treatment. There was a moderate positive correlation between stage of lactation and SCC (R2 = 0.438). PMID- 19294985 TI - Macroscopic features of the venous drainage of the reproductive system of the male ostrich (Struthio camelus). AB - The macroscopic features of the venous drainage of the reproductive system of the male ostrich were studied in six pre-pubertal and three sexually mature and active birds. Each testis was drained by one to four testicular veins. The right testicular veins drained the right testis and epididymis and its appendix to the caudal vena cava and to the right common iliac vein, whereas the left testicular veins drained the left testis and epididymis and its appendix exclusively to the left common iliac vein. A number of variations in the drainage pattern based on the point of entry and number of testicular veins were observed. The cranial aspect of the testis was also linked to the caudal vena cava or common iliac vein via the adrenal veins. The cranial, middle and caudal segments of the ductus deferens (and ureter) were drained by the cranial, middle and caudal ureterodeferential veins respectively, to the caudal testicular veins, the caudal renal veins and pudendal/caudal part of the internal iliac veins. In some specimens, the caudal ureterodeferential veins also drained into the caudal mesenteric vein. The surface of the phallus was drained by tributaries of the pudendal vein. The basic pattern of venous drainage of the reproductive organs of the male ostrich was generally similar to that described for the domestic fowl. However, important differences, including the partial fusion of the caudal renal veins, drainage of the cranial aspect of the testes via the adrenal veins, drainage of the caudal ureterodeferential veins into the caudal mesenteric vein and the presence of veins draining the surface of the phallus, were observed. Although significant, these differences may simply reflect variations in the normal pattern of venous drainage of the reproductive tract of birds which could be verified by studying more specimens and more species. PMID- 19294986 TI - Evaluation of larvicides in developing management guidelines for long-term control of pest blackflies (Diptera: Simuliidae) along the Orange River, South Africa. AB - In 2000 and 2001 Orange River levels were higher than normal: associated serious outbreaks of blackfly had a substantial detrimental impact on the local economy. The poor control was attributed to the suspected development of larval resistance to temephos. A long-term solution to blackfly control, through the identification of a suitable replacement to temephos for use during high flow conditions, was proposed. This study, however, failed to identify or register a suitable larvicide for use during high flow conditions. Although permethrin was highly effective against blackfly larvae, it was rejected because of its detrimental impacts on non-target fauna. Various formulations of locally produced dry Bacillus thuringiensis var. israelensis (B.t.i.) were tested, but these were ineffective against blackflies. The study also confirmed that resistance to temephos has developed among Simulium chutteri in the middle and lower Orange River. The feasibility of "reversing" the resistance to temephos through the use of the synergist piperonyl butoxide (PBO) was investigated, but the results were not favourable. Furthermore, PBO was highly toxic to blackflies and non-target organisms, and was not recommended for further testing. This means that B.t.i. currently remains the only symptomatic measure of treatment currently applied. Although resistance to B.t.i. has not been reported for blackflies elsewhere in South Africa, there is a need to remain vigilant and to implement an operational strategy that minimizes the risks of resistance developing. PMID- 19294987 TI - Prevalence of the dog nematode Spirocerca lupi in populations of its intermediate dung beetle host in the Tshwane (Pretoria) Metropole, South Africa. AB - Spirocerca lupi (Spirurida: Spirocercidae) is a cosmopolitan parasite, principally of domestic dogs and dung beetles are its main intermediate hosts. In South Africa there has recently been growing concern over the upsurge of reported cases of clinical spirocercosis in dogs, while little is known or understood about the dynamics of the host-parasite associations between dung beetles and this nematode. We determined and compared the prevalence of infection in dung beetles between rural, urban and peri-urban areas of Tshwane (Pretoria) Metropole. Dung beetles were sampled during April and October 2006, at various localities in each of these areas. Localities were selected on the basis of being focal areas of high infection with S. lupi in dogs. Pig, dog and cow dung-baited pitfall traps were used for sampling the beetles. Trap contents were collected 48 h after the traps had been set and only dung beetles were collected from the traps. In total, 453 specimens belonging to 18 species were collected from 63 pitfall traps in all three areas. The numbers of species that were collected varied among the three areas. Dung beetles, irrespective of species (18) and numbers (447), predominantly preferred pig dung. The prevalence of dung beetles infected with the larvae of S. lupivaried considerably in the three areas. In the urban area 13.5% of the dung beetles dissected were infected, while the prevalence of S. lupi in dung beetles in the rural area was 2.3%. All the dung beetles that were infected with this nematode showed a preference for omnivore (pig and dog) dung. PMID- 19294988 TI - Tomid parasites in fish in the Olifants and Comati River Systems, South Africa. AB - During parasitological field surveys of freshwater fish, sebekiid and subtriquetrid pentastome larvae were recovered from the body cavity or swim bladder of several fish species from various localities in Limpopo and Mpumalanga Provinces, South Africa. Sebekia wedli was recovered from the body cavity of Marcusenius macrolepidotus (Mormyridae) from Flag Boshielo Dam, Limpopo Province, and Alofia sp. and Subtriquetra rileyi were found in the swim bladder of Oreochromis mossambicus (Cichlidae) from the Phalaborwa Barrage, Limpopo Province. The latter species was also collected from the swim bladder of O. mossambicus in dams in the Phalaborwa region and the Ga-Selati River, Limpopo Province. A single specimen of Sebekia okavangoensis was present in the body cavity of Clarias gariepinus (Clariidae) in a dam on a sugarcane farm in the Komatipoort region, Mpumalanga Province. Pentastomid infections in the Mormyridae and Clariidae represent new host records. PMID- 19294989 TI - Production trials involving use of the FAMACHA system for haemonchosis in sheep: preliminary results. AB - In three trials conducted on two separate farms the production of sheep treated for naturally acquired haemonchosis using the FAMACHA system of targeted selective treatment (TST) (i.e. to treat only those animals unable to manage unaided in the face of heavy Haemonchus challenge) was compared to that of suppressively drenched sheep in the same flock. As expected by the research team who developed and evaluated the FAMACHA system, TST did result in some loss in production. However, despite high levels of worm challenge in two of the trials and the fact that the comparison was with suppressive drenching which is not sustainable, the total effect was relatively small in relation to the important advantage of using the TST as regards reduced selection for anthelmintic resistance (AR). Concerning the sustainability of worm control, it is concluded that the development of drug resistance to anthelmintics leaves sheep and goat farmers in South Africa no choice but to use methods of TST such as FAMACHA. The FAMACHA system can also be a useful clinical aid for early on-farm detection of AR by farmers; the degree of improvement in the colour of the ocular mucous membrane from pale to red in individually drenched anaemic animals over a period of 7-14 days can give a good indication of the efficacy of the compound(s) used. PMID- 19294990 TI - A rapid and sensitive real-time reverse transcription PCR for the pathotyping of South African H5N2 avian influenza viruses. AB - A Fluorescence resonance energy transfer (FRET) real-time reverse-transcription (rRT-PCR) assay was developed that distinguishes stains of South African and European highly pathogenic (HPAI) from low pathogenicity (LPAI) H5 avian influenza viruses in the absence of virus isolation, irrespective of the length of insertion at the hemagglutinin cleavage site (H0). The assay was used to pathotype H5-type viruses detected by rRT-PCR in ostrich tracheal swabs collected during the 2006 HPAI H5N2 outbreak in the Western Cape Province. PMID- 19294991 TI - Sequence and phylogenetic analysis of chicken anaemia virus obtained from backyard and commercial chickens in Nigeria. AB - This work reports the first molecular analysis study of chicken anaemia virus (CAV) in backyard chickens in Africa using molecular cloning and sequence analysis to characterize CAV strains obtained from commercial chickens and Nigerian backyard chickens. Partial VP1 gene sequences were determined for three CAVs from commercial chickens and for six CAV variants present in samples from a backyard chicken. Multiple alignment analysis revealed that the 6% and 4% nucleotide diversity obtained respectively for the commercial and backyard chicken strains translated to only 2% amino acid diversity for each breed. Overall, the amino acid composition of Nigerian CAVs was found to be highly conserved. Since the partial VP1 gene sequence of two backyard chicken cloned CAV strains (NGR/CI-8 and NGR/CI-9) were almost identical and evolutionarily closely related to the commercial chicken strains NGR-1, and NGR-4 and NGR-5, respectively, we concluded that CAV infections had crossed the farm boundary. PMID- 19294992 TI - Epidemiological survey on gastro-intestinal and blood-borne helminths of dogs in north-east Gabon. AB - A survey of helminth parasites was carried out on 198 dogs living in almost complete liberty in villages in the northeast of Gabon. Faeces and blood samples were collected and analysed. Dirofilaria immitis antigen was detected in 13.6% of dogs using the SNAP 3Dx test, a commercially available enzyme-linked immunosorbent assay (ELISA). Faecal examination revealed that 91.4% of dogs were infected by intestinal helminths. Ascarids were found in 58.5% of the samples. Trichuris vulpis was observed in 49.5% of cases, and Uncinaria spp. and Ancylostoma spp. in 34.8%, Spirocerca lupi in 25.3% and Capillaria spp. in 10.6%. Cestode embryophores were found in 8.6% of the samples. PMID- 19294993 TI - [Resilience and precarity]. PMID- 19294994 TI - [Heart insufficiency in elderly--new dimensions of the problem]. AB - The 21st century's demographic and epidemiologic perspectives suggest an absolute and percentual increment of elderly population. In the same time the prevalence of heart insufficiency increases with advanced age, which transforms heart failure in a true challenge for health insurance system. This new dimension of the problem resides in the increment of heart insufficiency's prevalence and in the high costs of medical care of these patients. Heart failure's medical costs are high not only due to the necessity of compensating the cardiac function but also due to the necessity of treating the etiology, the risk factors and the co morbidities with cardiac side effects. Elderly patients usually have a low therapeutic compliance and an important number of iatrogenic reactions mainly due to poly-medication, poly-pathology and cognitive and physical deficiencies, which need special measures of therapeutic education for the patient and family and medical survey, probably best in a nursing house; all these elements increase significantly the cost, which is almost double than the cost for cancer. The prophylaxis in heart insufficiency means an extremely efficient management of entire cardiac pathology in order to delay the moment of heart failure. In elderly patients, the treatment in heart insufficiency is difficult not due to hospital treatment of decompensation episodes but due to low therapeutic compliance at home. We need special programs to ensure medical education of the patient and the family/caregiver in order to avoid iatrogenic pathology and to ensure therapeutic efficiency. PMID- 19294995 TI - [Refractory arterial hypertension]. AB - The present review synthesizes "the state of the art" concerning resistant (refractory) arterial hypertension (RAH). Definition, epidemiology, frequent risk factors, clinical conditions and causes of therapeutic resistance of RAH are revisited. Pathogenic hypothesis and related therapeutic approaches are explored. The paper lists current and new pharmacological therapies of RAH and introduces the latest interventional approach (the Rheos BHT system) to this form of arterial hypertension. PMID- 19294996 TI - [Involvement and role of iron in nonalcoholic steatohepatitis]. AB - Nonalcoholic steatohepatitis (NASH) was described by Ludwig mainly in obese, middle-aged women, often associated with diabetes mellitus and hyperlipidemia. In the recent years, NASH was found to be associated with male, nonobese, nondiabetic patients and with liver iron overload, which led to the hypothesis of iron playing a role in NASH pathogenesis. Increased ferritin with normal transferrin saturation is frequently found in fatty liver patients, but it reflects iron overload only in those patients in which it persists despite an appropriate diet. Insulin resistance hepatic iron overload (IR-HIO) is a new condition of hepatic iron overload, characterized by hyperferritinemia with normal or slightly increased transferrin saturation in the absence of hemochromatotic gene mutations. Although patients with IR-HIO have a high prevalence of insulin resistance-related metabolic disorders, the relationship of IR-HIO and NASH is unclear. Two characteristics allow differentiation of IR-HIO from genetic haemochromatosis: iron overload is heterogeneous from one hepatocyte to another in the periportal area, and sinusoidal iron is distributed throughout the lobule. In IR-HIO, fibrosis develops at a much lower hepatic iron burden than in genetic haemochromatosis, and sinusoidal iron, steatosis and inflammation could represent the histological mark of activity and progression of liver disease in IR-HIO. PMID- 19294997 TI - [Hemostatic disorders in liver disease]. AB - Hemorrhagic complications are common in patients with liver diseases and contribute to the morbidity and mortality associated to this condition. The liver plays a central role in the hemostatic process as here all clotting factors and their inhibitors are synthetized. Liver damage is commonly associated with variable impairment of hemostasis due to multiple causes: decreased synthesis of clotting and inhibitor factors, decreased clearance of activated factors, hyperfibrinolysis, accelerated intravascular coagulation, quantitative and qualitative platelet defects. Their clinical implications remain to be elucidated, so further studies addressing this issue are needed. PMID- 19294998 TI - [Alcoholic ketoacidosis]. AB - Alcoholic ketoacidosis is an acute metabolic acidosis that typically occurs in people who chronically abuse alcohol and have a recent history of binge drinking, little or no food intake and persistent vomiting. Alcoholic ketoacidosis is a result of starvation with glycogen depletion and counter-regulatory hormone production, a raised nicotinamide adenine dinucleotide (NADH) to nicotinamide adenine dinucleotide (NAD+) ratio related to the metabolism of ethanol, and volume depletion resulting in ketogenesis. Alcoholic ketoacidosis is characterized by elevated serum ketone levels and a high anion gap. Once the diagnosis of alcoholic ketoacidosis is made, the mainstay of treatment is hydration with 5% dextrose in normal saline. With timely and aggressive intervention, the prognosis for a patient with alcoholic ketoacidosis is good. PMID- 19294999 TI - [Psychogenic pain in children]. AB - Children present many relations between psychic and somatic processes. The most important clinical aspect is the pain that cannot be attributed to any medical condition and occurs in at least one organ. Psychogenic pain is frequent in the pediatric population and its incidence is increasing. The authors present some of the most debated theories on the mechanisms of psychosomatic pain and some types of such pain, underlining the importance of full investigations. The pain is diagnosed as psychogenic after the exclusion of a potentially lethal disease. PMID- 19295000 TI - [Prognostic value of brachial artery flow--mediated dilation and carotid artery intima-media thickness in hypertensive patients]. AB - Endothelial vasodilator dysfunction assessed by brachial artery flow-mediated dilatation (FMD) and carotid intima-media thickening (IMT) are two indicators of subclinical atherosclerotic disease. The aim of the study was to observe if there is a relationship between FMD and IMT in a group of hypertensive patients. MATERIAL AND METHOD: We measured FMD and IMT by ultrasound in 75 subjects aged between 42 and 62 years. We examined their correlation and interaction with traditionally cardiovascular risk factors. Endothelial function was assessed by means of flow mediated dilation (FMD) on brachial artery, using B-mode ultrasonography. RESULTS: The mean value of FMD was 8.00 +/- 2.02%. We observed a moderate but significant correlation between FMD and total cholesterol (r = 0.511, p < 0.01) and between FMD and triglycerides (r = -0.325, p < 0.01). All the patients in the study were also examined by high resolution B-mode ultrasound to measure the IMT of the common carotid artery. The mean value of IMT values was 1.41 +/- 0.3 mm. The correlation between FMD and IMT was inverse, moderate and significant. In conclusion, in hypertensive patients, B-mode ultrasound intima media thickness and FMD measurement are important tools for evaluation of arterial wall remodeling and can be used as valid markers of atherosclerosis in cardiovascular risk prediction. PMID- 19295001 TI - [The importance of carotid ultrasound in the risk stratification of patients with ischaemic heart disease]. AB - The stratification of cardiovascular risk is important both in primary and secondary prevention. In the clinical evaluation, besides the well-known risk factors, the European Guide of Cardiovascular Prevention recommends the identification of additional markers of risk, one of them being the detection of atherosclerotic alterations by carotid ultrasound examination. The objectives of this study were: assessment of carotid ultrasound alterations in patients with known cardiovascular disease; their relation with the other risk factors, the clinical status and echocardiography. METHOD: The study included 144 patients, aged 41-80 (84 males, 60 women) with cardiovascular disease (stable angina, myocardial infarction, peripheral arterial disease, arterial hypertension). Risk factors as smoking, hypercholesterolemia, diabetes mellitus, obesity have been determined, followed by a clinical assessment and echocardiographic examination. Ultrasound of the extracranial carotid arteries included measurement of IMT of the common carotid arteries, the identification of atherosclerotic plaques and the presence of stenoses. The plaques were considered if the IMT was more than 1.3 mm. RESULTS: Patients with IMT > 0.8 are older (61.5 +/- 8.2 vs. 56.1 +/- 8.2 years, p = 0.001); the prevalence of arterial hypertension is greater (63.8% vs. 21.6%, p = 0.05), and at an IMT of more than 1.1 mm all were hypertensive; the association with peripheral arterial disease is significant at the IMT = 0.8 mm (23.6% vs. 4.8%, p = 0.01). The relationship with echocardiographic markers of left ventricular hypertrophy is present at an IMT of 0.8 mm and is evident at IMT = 1 mm (interventricular septum 13.2 +/- 3 vs. 12 +/- 2.3, p = 0.05; left ventricular wall 14 +/- 5.4 vs. 12 +/- 1.7, p = 0.01; left ventricular mass 323 +/- 94 vs. 282 +/- 68, p = 0.03), and also the relation with the degree of diastolic dysfunction expressed by E/A ratio (0.71 +/- 0.24 vs. 0.99 +/- 0.39, p = 0.001). The presence of atherosclerotic plaques correlates with gender (45.1% at male vs. 24.3%, p = 0.01) and older age (61.7 +/- 8.6 vs. 56.2 +/- 8.5 years, p = 0.001). IMT and carotid plaques are not significantly correlated either with such risk factors as smoking, hypercholesterolemia, diabetes mellitus or clinical conditions as myocardial infarction and angina. CONCLUSION: IMT greater than 0.8 mm is a marker of increased cardiovascular risk, associated with age, presence of peripheral arterial disease, arterial hypertension, left ventricular hypertrophy, and diastolic dysfunction. IMT seems to be a more reliable index of cardiovascular risk than carotid plaques. Measurement of IMT is easy to be done in clinical practice, and is recommended for more accurate risk stratification in patients with atherosclerotic cardiovascular disease. PMID- 19295002 TI - [Nutritional status evaluation in maintenance hemodialysis patients]. AB - Protein-calorie malnutrition is a common complication and an important predictive factor for mortality in patients with end-stage renal disease on maintenance dialysis. Therefore, nutritional status needs to be regularly assessed in these patients by using several methods. If malnutrition is diagnosed, its causes should be thoroughly searched for and properly treated. MATERIAL AND METHOD: This cross-section study aimed at evaluating the nutritional status and the possible risk factors for malnutrition in 149 (82 men) hemodialysis patients by anthropometry, biochemical tests and bioelectrical impedance analysis (BIA). The patients' height (H), post-dialysis body weight (BW), mid-arm circumference (MAC), tricipital skin-fold thickness (TST) were measured and a 3-category subjective global assessment (SGA) was performed. Body mass index (BMI), mid-arm muscle circumference (MAMC), corrected mid-arm muscle area (cMAMA) and anthropometry-estimated percent body muscle mass (% AMM) were calculated from the above measurements by using specific equations. Biochemical tests included protein equivalent of nitrogen appearance (nPNA), and predialysis serum albumin, creatinine, total cholesterol, bicarbonate, and hemoglobin (Hb) levels. We used BIA to estimate body composition - i.e. percent body fat (% BBF), fat-free mass (% FFM), body cell mass (% BCM), extracellular mass (% ECM), muscle mass (% BMM)- and the phase angle (PhA). T-test was used to make comparisons and Pearson coefficient to analyze the correlations. P < 0.05 was considered statistically significant. RESULTS: The male patients had a higher mean muscle mass--as estimated by serum creatinine (9.8 s 8.3 mg/dl; P < 0.0001) and by % BMM (41.7% vs 34.7%)--and a lower fat mass--as estimated by TST (0.95 cm vs 1.2 cm; P = 0.016) and by % FAT (16.7% vs 31.3%; P < 0.0001) than the female patients. Age was found to be positively correlated with BMI (P = 0.001), but inversely correlated with % BCM (P = 0.013) and with % AMM (P = 0.003). Patients with diabetes had lower % BCM than those without diabetes (32.9 vs 35.9%; P = 0.041). The presence of heart failure was associated with significantly reduced MAMC (22.0 vs 23.6 cm2; P = 0.045), % AMM (28.5 vs 32.1; P = 0.021), % BCM (33.0 vs 36.1% ; P = 0.034), BMM/H2 (8.6 vs 9.4 kg/m2; P = 0.013), nPNA (1.17 vs 1.34 g/kg d ; P = 0.047), serum albumin (39.7 vs 42.4 g/l; P = 0.010), serum creatinine (8.1 vs 9.4 mg/dl; P = 0.008) and Hb (10.5 vs 11.2 g/dl; P = 0.017). The serum Hb level was positively correlated with BMI (P = 0.005), BMM/H2 (P = 0.009), serum albumin (P = 0.002) and serum creatinine (P = 0.011). Also, patients with category B-SGA were older (63.7 vs 50.1 y.o.; P < 0.0001) and had more heart failure (42% vs 13%; P = 0.013) than those with category A-SGA. In hemodialysis patients, advancing age, diabetes, heart failure and decreasing Hb levels are associated with worse nutritional status, as estimated by anthropometry, biochemical markers and BIA. Whether treatment of comorbidities such as heart disease and anemia may improve nutritional status in these patients is an important issue that deserves further research. PMID- 19295003 TI - [Celiac disease: diagnostic criteria and impact of gluten free diet--patients' perspective]. AB - Clinical, serological and histological remission in celiac disease is strictly related to long life complete exclusion of gluten from the diet. Because celiac disease is an autoimmune systemic disorder the potential complications include a wide spectrum with two severe long-term morbidities: osteoporosis and malignancy. Gluten free diet, easy to be recommended, is difficult to be followed. This study aimed at evaluating the diagnostic criteria and dietary instruction, quality of life issues, impact of the gluten free diet on the celiac patient. MATERIAL AND METHOD: 250 patients (mean age at diagnosis 18.18 years) on gluten-free diet were asked to fill out a disease specific questionnaire. RESULTS: The response rate was 58.4% (mean age at diagnosis 11 years); 7% had no biopsy proven diagnosis (being excluded) and 28.26% had only antigliadin antibodies tested as serology method; 84.78% had typical digestive symptoms at diagnosis; 28.2% had been followed up after the diagnosis; information about the disease and diet have been provided exclusively by the gastroenterologist in 95.6% of the responders, 45.6% considering the information insufficient. The gluten free products were considered in 73.9% of cases too expensive for the family income, with low availability, lack of variety and unclear labeling; 48.8% reported impaired access to certain activities (social events, traveling) and even having difficulties to be accepted for some jobs. CONCLUSIONS: There is evidence that an educational program is needed to improve knowledge on the atypical forms of disease, diagnostic criteria and tools. The quality of life would be improved by support groups for patients, improved availability, better prices and food labeling. PMID- 19295004 TI - [Digestive disease in the immunocompromised patient with acute lymphoblastic leukemia. Experience of the IVth Pediatric Clinic--Oncology Department of the Iasi Sfanta Maria Children's Hospital]. AB - The goals of this paper were to study the various types of digestive disease in acute lymphoblastic leukemia (ALL), to characterize the children in the study group by age, sex and environment, by presence of liver and spleen enlargement, levels of GOT, GPT, vomiting, to evaluate methotrexate (MTX) serum levels al 24 hours and 48 hours after administration, and to analyze the correlation between MTX levels and MTX liver and blood toxicity. MATERIAL AND METHOD: We studied 39 immunocompromised children hospitalized in the IV-th Pediatric Clinic-Oncology Ward, between 1983-2005, with acute lymphoblastic leukemia (ALL); most of them exhibited defects of humoral immunity such as transitory hypogammaglobulinemia, and defects of the cellular immunity that accompanied hepatomegaly, hepatic cytolysis and biliary obstruction. RESULTS: The diagnostic of ALL was sustained by: medullar biopsy, lumbar punction, cytochemical reactions, blood cell count, flow-cytometry, methotrexate level determination. Hepatic damage was measured by: GOT, GPT, bilirubin, LDH, GGT, FA, HBS antigen, anti HCV antibodies, anti HVA antibodies, anti toxoplasmosis antibodies, anti CMV antibodies, serum protein levels, TQ, inflammation markers. A slight increase in the number of diagnosed ALL cases during the past two decades was noticed and ALL incidence was higher in boys than girls. Hematologic toxicity of MTX is a real problem, causing neutropenia. MTX also caused oral lesions (in 69.36% of children), vomiting (in 69.2% of children) and liver toxicity (in 51.3% of children). MTX serum level 24 hours after administration is significantly different from the serum level at 48 hours after administration. Thus, the use of calcium folinate is proved to be very effective. We have also demonstrated that vomiting had some other causes besides MTX administration. PMID- 19295005 TI - [Clinical and prognostic considerations in thalamic hemorrhage. Clinical study on 117 cases]. AB - Intracerebral hemorrhage located in the thalamus is one of the most frequent forms of hemorrhagic stroke in hypertensive patients. In this study the authors investigated the most important aspects related to thalamic hemorrhage : risk factors, initial signs, neurologic and EEG findings, the prognostic impact of the presence or absence of ventricular penetration, as well as the extension of hematoma. MATERIAL AND METHOD: 117 patients with thalamic hemorrhage confirmed by cerebral computerized tomography were included in the study. RESULTS: Hypertension was the most significant risk factor in our patients. Headache was the most common early clinical symptom, while pyramidal signs were the most frequent findings at neurological examination. Patients with hemorrhagic extension to the putamen had the worst prognosis for stroke. PMID- 19295006 TI - [The management of unexplained recurrent abortions. Is immunotherapy an option?]. AB - Recurrent abortions, defined as more than three consecutive pregnancy losses, are associated with genetic, anatomic and endocrine causes. Whenever investigations fail to detect a cause, immunological dysfunctions are incriminated. The paper shortly reviews the main causes, investigation protocols and immune therapeutic attempts that have been made. Active immunization with allogenic leukocytes or trophoblastic membranes, passive immunization with intravenous immunoglobulins and immunomodulation with glucocorticoids in unexplained recurrent abortions generated contradictory results and was banned by reproductive immunology scientists as empirical, risky for the mother and harmful for the fetus. Confronted with desperate couples, clinicians use the immunotherapy with paternal lymphocytes or intravenous immunoglobulins in healthy women with unexplained recurrent abortions or in cases where accepted therapies failed. PMID- 19295007 TI - [The value of surgical therapy in cervical nodal metastases]. AB - Aside from distant metastasis, cervical nodal metastases are the worst prognostic indicator for survival in patients with head and neck carcinoma. Presence of nodal metastases decreases survival by fifty percent. This paper refers to surgical therapy of cervical adenopathy known as the radical neck dissection, for clinically positive nodal metastases, follow by chemotherapy or radiotherapy depending on the primary tumor site. PMID- 19295008 TI - [The psycho-social impact of the locomotor deformations in school-aged child]. AB - Depending on their severity, congenital or acquired, the deformations of the locomotor system affect more or less the child's mental status. Adolescence is the time when their impact is most important. When the physical, mental or functional suffering become a disability, professional and psycho-social integration problems occur. The role of the team treating such a patient is to orientate him toward a specific job corresponding to his abilities, while the social services must support him in achieving this goal. The psychotherapist is the most important person in his fight against depression and, more important, suicide. PMID- 19295009 TI - [Abdominal wall endometriosis after gynecologic procedures: an under-appreciated diagnosis in general surgery]. AB - Incisional endometriosis is a clinical entity described in the gynecologic literature but it is not well recognized among general surgeons. The preoperative diagnosis is often mistaken for a suture granuloma, abscess, lipoma, cyst or incisional hernia. MATERIAL AND METHOD: This is a retrospective review of six cases of incisional endometriosis in our hospital aimed at determining which, if any, factors would suggest the diagnosis preoperatively. All general surgery patients with a diagnosis of endometriosis in their pathology specimens from January 1990 to February 2006 were reviewed. RESULTS: All six patients had previous cesarean sections through either a Pfannenstiel (n = 4) or lower midline (n = 2) incision. Ages ranged from 28 to 43 years (mean 34.3 years). All patients presented with a palpable painful lesion located in the area of cesarean section incision. Three of the patients had a change in symptoms with their menstrual cycle. The duration of symptoms ranged between 2 months to 6 years. All patients underwent surgical excision. The size of the excised endometrioma ranged from 3 cm to 9 cm (mean 5.3 cm). CONCLUSION: Incisional endometriosis seems to be common in women with a history of cesarean section. Most patients presented with a painful abdominal mass. Three patients presented cyclic changes in pain and size of the mass with menses, but this may be due to physician's lack of awareness and questioning. The management of choice is to remove the lesion completely even if fascial excision is required. PMID- 19295010 TI - [Neurological and abdominal lesional associations in polytrauma]. AB - Polytraumatisms have always been a concern in human history, their number increasing especially during wars. Their forecast depends on many factors, starting from means of transportation, first aid's promptitude, technical endowment which evolved a lot within the years. Amongst the thoracal-abdominal, the cranial-cerebral and vertebral-medullary injuries, the cranial-cerebral and the abdominal injuries are the most severe ones. We hereby present the evaluation scales according to various criteria (seriousness, prognosis, clinical valutation). In the case of abdominal and neurosurgical injuries the symptomatology can be modified by addition or defalcation, which may cause diagnostic errors with consequences upon morbidity and mortality. MATERIAL AND METHOD: We hereby present a study accomplished within three departments: Piatra Neamt Emergency Hospital, Piatra Neamt Coronary Office and Iasi Neurosurgery Clinic, and we find the injury associations in polytrauma cases. RESULTS: We notice a prevalence of the male sex, of the country people, of traffic accidents, of the active age 20-60 years old. The association within abdominal trauma and neurological injuries is of 16.97% in Iasi Neurosurgery Clinic. After being solved, the neurological injuries are sent to surgery departments, for solution or surveillance. CONCLUSION: In order to optimize the medical assistance for traumatized persons we hereby recommend a polytrauma centre establishment. PMID- 19295011 TI - [Struma ovarii]. AB - Struma ovarii is a rare tumor in its pure form, but its true incidence is hard to estimate because of all the variation in the diagnostic criteria due to the fact that some authors reported it as within a teratoma and others only in its pure form. In general, it is an asymptomatic tumor, "benign-like" in most of the cases, and the diagnostic is based only on the histopathological findings. MATERIAL AND METHOD: This is a retrospective study of all ovarian tumors treated in our departments over a period of 11 years, from January 1, 1995 to January 1, 2006. RESULTS: All medical records of 644 patients were reviewed, of which 502 patients (77.96%) with unilateral tumors and 142 (22.04%) with bilateral involvement. 879 tumors were removed: 88.39% benign and 11.61% malignant. Struma ovarii was demonstrated in 2 patients. The only complaint in these 2 patients diagnosed and treated in our clinics was pain in the lower abdomen. The clinical examination and ultrasound suggested ovarian tumors, but the diagnosis was made on the histopathological findings. Following surgical treatment (classic or laparoscopic) the postoperative course was excellent, confirming that this approach is the only practical solution. PMID- 19295012 TI - Open diverticulectomy in a patient with a giant symptomatic retro-pancreatic duodenal diverticulum. A case report. AB - Although often incidentally found, diverticular disease of the small bowel and duodenum may generate complications and impose surgical treatment. Diagnosis of small bowel diverticulosis is difficult because the clinical picture is usually ambiguous. We report a case of diverticulosis of the whole intestine including a giant symptomatic retro-pancreatic duodenal diverticulum in a 74-year-old patient who presented with recurrent episodes of abdominal pain, weight loss and intestinal obstruction. The diagnosis was made by MRI enteroclysis and endoscopy. Open diverticulectomy with choledochostomy and insertion of a T-tube was performed and resulted in a rapid improvement of the symptoms. PMID- 19295013 TI - [Splenic infarction in endocarditis: case report]. AB - Splenic infarction is a very rare event though it can occur in a multitude of conditions with general or local manifestations. Splenic involvement in infectious endocarditis can be either infarction or abscess formation. The clinical picture is usually nonspecific and the diagnosis is often not initially suspected. We present a case of a 67 year-old male patient with infectious endocarditis who has been diagnosed with a splenic infarction by abdominal CT scan. The infarction was a result of septic embolization from the infected endocardium. The patient was initially managed conservatively with triple antibiotic therapy and regular follow-up ultrasound scan. The persistence of abdominal symptoms and the suspicion of splenic abscess formation led to the decision of performing splenectomy. Although imaging can characterize the nature of a splenic lesion, it is sometimes impossible to differentiate between infarction and abscess without histopathological confirmation. Splenic infarction should be suspected in all patients with a past history of thromboembolic disease who complain of left upper quadrant pain and present with localized or systemic inflammatory signs. Unfavorable clinical course and suspicion of abscess formation are indications for surgery. The characteristic features of splenic infarction are discussed together with a review of the recent literature. PMID- 19295014 TI - [Cystic mucinous tumors of the pancreas]. AB - Mucinous cystic tumors (MCTP) are uncommon borderline lesions of the pancreas that may often be misdiagnosed as pseudocysts and have unknown propensity to malignity. During the last two decades in our clinic of 38 patients with cystic lesions of the pancreas two presented MCTP: a 48-year-old woman with a cyst located in the body and tail of the pancreas and a 59-year-old man with a tumor situated in the head of the gland producing an obstructive jaundice. Ultrasonography and computer tomography confirmed the presence of the cystic mass in both patients but the precise diagnosis was made only after the histologic examination of the removed lesions. With an attempted curative resection: distal pancreatectomy with preservation of the spleen in the first case and a Whipple procedure in the second one, both cases were cured. Pathology do not reveal malignant transformations. Both patients were alive, had no complaints or recurrences at 12 and 24 month after surgery. The treatment of choice in such cystic tumors is surgical removal as untreated these lesions may progress to neoplastic degeneration. PMID- 19295015 TI - [Primary total hip arthroplasty in bilateral acetabular protrusion, case report]. AB - Acetabular protrusion is a clinical entity consisting of deepening of the acetabulum and consequent sinking of the femoral head within this cavity, uni- or bilaterally, such that the bottom of the acetabulum protrudes in the pelvic cavity. Etiologically, the disease may be primary or secondary. This article presents the case of a 66-year-old man with primary bilateral acetabular protrusion admitted to our clinic for symptoms involving the hips: pain exacerbated at big effort in erect position and a progressive diminution of the amplitude of movement in both hips. The plain radiograph of the pelvis revealed the deepening of the acetabulum on both sides, intrapelvic protrusion of the bottom of the acetabular cavities, sinking of the femoral heads (and to a certain extent, also the femoral necks) into the acetabula. The articular space was altered bilaterally by osteoarthritic modifications. The diagnosis was bilateral acetabular protrusion with secondary hip osteoarthritis. Subsequently, he underwent bilateral total hip arthroplasty with an additional reinforcement with a Burch-Schneider ring on the left acetabulum. The operations were performed at a 6-months interval, with a favorable postoperative course for both hips. PMID- 19295016 TI - Correlation between E-cadherin abnormal expressions in different types of cancer and the process of metastasis. AB - Cell-cell adhesion plays a critical role in the establishment and maintenance of cell polarity and cell society. Reduced cell-cell adhesiveness allows cancer cells to disobey the social order, resulting in destruction of the histological structure, the morphological hallmark of malignant tumors. Morbidity in most cancer patients is not due to primary cancer but to metastatic disease. Thus, understanding the progression of tumors to metastatic state and the changes that take place in highly aggressive cells is important in the development of novel approaches to the diagnosis and treatment of progressive malignancies. Cell adhesion molecules are implicated in human carcinogenesis. E-cadherin is a calcium-dependent cell adhesion molecule the intact function of which is crucial for the establishment and maintenance of epithelial tissue polarity and structural integrity. The gene encoding E-cadherin (CDH1, on chromosome 16q22.1) was one of the first to be considered as an invasion-suppressor gene. Mutations in CDH1 occur in diffuse type gastric cancer, lobular breast cancer, and endometrial cancer. In human cancers, partial or complete loss of E-cadherin expression correlates with malignancy. Through immunohistochemical analysis it has been assessed the abnormal expressions of E-cadherin in three types of cancer: gastric carcinoma, lobular breast carcinomas and cutaneous melanoma and the correlation with the multistep process of metastasis. PMID- 19295017 TI - [Contribution of immunohistochemistry and flow-cytometry in the study of endometrial pathology]. AB - Endometrioid endometrial carcinoma is characterized by hyperestrogenism and high sensitivity to progestogens. Our study was designed in order to demonstrate the hormonal influences in neoplastic endometrium, by investigating steroid receptors expression and their correlation with proliferation activity, with matrix enzymes expression, and with susceptibility to apoptosis inducers, from hyperplasia to carcinoma. 13 cases were included in our study, 7 endometrial hyperplasias, and 6 endometrial carcinomas. Immunohistochemistry technique was performed, using antibodies against ER-a, PR, PCNA, MMP-2, MMP-9, Fas, and FasL molecules. Flow cytometry was complementary used, for steroid receptors, PCNA and MMPs. Endometrial hyperplasias were positive for both hormonal receptors, in epithelial and stromal cells. An evident decrease of the percent of positive cells and of the staining intensity of both ER and PR was observed in poorly differentiated endometrial carcinomas. Endometrial hyperplasias presented a similar proliferative index with differentiated endometrioid endometrial carcinomas. Poorly differentiated endometrial carcinomas showed the highest PCNA index. Both types of investigated MMPs were evident, with similar aspect of localisation for both MMP-2 and MMP-9. The staining intensity for MMP-9 was higher than that of MMP-2, and was identified both in epithelial and in stromal cells. Fas and FasL expressions were identified in glandular epithelium of endometrial hyperplasias and carcinomas, although the staining intensity was reduced. Flow-cytometry showed a correlation between qualitative and quantitative data concerning hormonal receptors, PCNA and MMPs. Our study emphasises that neoplastic endometrial cells express several molecules correlated with malignant transformation and tumoral progression, by coordinated intervention of steroids, proliferating factors, gelatinases, in opposition with systems involved in apoptosis initiation. PMID- 19295018 TI - [Experimental study regarding the ureteral cicatrization pattern in rabbit]. AB - This study aimed to elaborate an experimental model for ureteral cicatrization following surgical lesion, with direct involvement in urological therapy. MATERIAL AND METHODS: The study was realized on a group of 9 female rabbits on which we have performed ureteral surgery. First surgical event consisted in partial cut of the left ureter following transperitoneal approach, ureteral stent insertion and the suture of the ureteral wound. The second surgical event has accomplished the harvesting of the ureteral fragment during cicatrisation process and was performed at various times from the first surgical event, accordingly at 1, 2, 3...6 days--on different rabbits. 3 rabbits composed the witness group. Fragments were fixed in formol 4%, and histologically stained with hematoxilin eosin and van Gieson. RESULTS: In the first two days we have observed an obvious inflammatory process on the postoperative ureteral scar. In days 3 and 4, the limited fibrosis appeared in the 2nd day engaged a peak in the 4th day when appeared a sketch of ureteral lumen constriction. In the 5th and 6th day the fibrosis process underwent a moderate resolution, simultaneously with a local diffuse congestion, marker for the remodeling processes of the connective matrix. CONCLUSION: Animal cicatrisation model follows the same pattern as in human but at different timing so as extrapolation requires considering these facts. PMID- 19295019 TI - [Considerations regarding the treatment of the algodystrophy using the acupuncture]. AB - MATERIAL AND METHOD: A team made up of 14 subjects with algodystrophy (AD) was studied at the level of the distal zone of one superior member result of the radius and/or ulna fracture, which needed an immobilization device for 30-40 days, using acupuncture. RESULTS: The results were compared with those obtained at a witness team treated by electro-physical recovery (ultrasound-therapy, ultrasound-phoresis with hydrocortisone, Bernard diadynamic currents and physiotherapy). Taking into account that AD is the local expression of the General Syndrome of Adaptation, the acupuncture method was used for the improvement of the integration function of the conjunctive tissue. CONCLUSION: Through the treatment with acupuncture the treatment period is much shorter, and the movement functions are much better. PMID- 19295020 TI - [New social and economical issues in the assessment of Romanian population's health status]. AB - Health status is determined by a combination of biological, environmental (physical and socio-economic), behavioural/lifestyle and medical care factors. The social and economic factors include many influences over which an individual may have limited control, such as economic status or educational level. The most important measures of these factors are represented by: gross domestic product per capita, employment rate, unemployment rate, literacy rate, poverty line, and human development index. From this point of view, the following positive issues have been recorded in Romania: a low unemployment rate (6.1%) compared to European Union countries; a high literacy rate (97.3%), very close to the maximum of 100% reported by all developed countries; and a human development index of 0.805 (rank 60 in the international hierarchy), specific to a country with a high human development. Negative issues have been reported in case of the following indicators: the reported gross domestic product per capita expressed in PPP US$ was 8480, among the lowest in Europe, specific to a country with a medium income; population living with less than 2 US$ per day of 13% and living with less than 1 US$ per day 2.1%; the employment rate was 57.4%, but in female population only 51.3%, whereas in male population it was 63.9%. PMID- 19295021 TI - [The costs of extreme values and case resolved in a public hospital from Iasi, Romania]. AB - For calculating the estimated costs for health care services in public hospitals from Romania case mixed index and mean of hospital duration are used especially. Medical insurance give for medical practice a fixed allowance based on a historical cost. For hospitals with severe cases there is necessary to introduce the term "extreme values, or cases" for improving the cost for intensive care units, for complicated cases. PMID- 19295022 TI - [Epidemiological assessment of morbidity and natural foci of TBE-CEE virus infection in Transylvania]. AB - In Romania, anthropozoonoses, and in this nosological context, human viral neuro infections with incidental transmission by active vectors with natural foci, may be considered infectious diseases with endemic emergence. Since 1996, the form of manifestation of the epidemiological process by viral meningitis and meningoencephalitis, the etiological structure of morbidity and the problem of natural foci have required the implementation of an effective epidemiological survey strategy. This paper includes the results of an epidemiological survey and the general population seroepidemiological evaluation of acute and historic infections, identified as arboviral TBE-CEE neuro-infections. MATERIAL AND METHOD: In the interval 1999-2006, the following aspects were studied: an endemic episode of 37 cases of TBE-CEE neuro-virus infection in a rural population. The epidemiological inquiry was conclusive for the transmission of infection by goat milk from highly tick infested animals, in a habitat with identified natural sylvatic and peridomestic foci; an endemic-epidemic state with 17 cases of TBE CEE neuro-virus infection, occurring within a short time interval, in a rural community. Recent history revealed tick infestation from a common natural area; the size and dynamics of territorial changes in natural foci by comparing the prevalence of historic TBE-CEE infections in 41 subjects at known occupational risk, 95 subjects from the domestic habitat of former patients with TBE-CEE neuro infection, and 172 subjects from areas adjacent to natural foci. RESULTS: Specific serology was positive in 41.5%, 11.6%, and 5.8% of the investigated subjects (chi2 = 38.98); the prevalence of historic TBE-CEE infection in the general population was estimated at 0.6% (0.3-1.1%, CI = 95%) by the seroepidemiological investigation of a representative sample of the Transylvanian population (1669 subjects); the TBE-CEE etiology of acute viral meningitis/meningoencephalitis in the interval 2001-2006 (862 cases in the area) with a prevalence of 14.1% (10.9-17.8%, CI = 95%) of the cases, evaluated based on the etiological investigation of representative samples from 2 counties and on request from the territory (in total 397 investigated cases). Seroepidemiological tests validated by external quality control were performed using the ELISA technique, with standardized immune reagents (IgM and/or IgG specific). CONCLUSION: Based on the epidemiological survey performed, it may be concluded that the specific infection sources and the morbidity of human TBE-CEE virus neuro-infections have an endemic emergent course and natural foci are in full territorial expansion. PMID- 19295023 TI - [Antimicrobial resistance profile of E. coli and Klebsiella spp. from urine in the Infectious Diseases Hospital Iasi]. AB - The increasing frequency of extended-spectrum beta-lactamases (ESBLs) producing Enterobacteriaceae among nosocomial and community-acquired infections is an important problem for both microbiologists and clinicians, because of the difficulty in correctly detecting, reporting and treating such infections. RESULTS: In the Clinical Hospital of Infectious Diseases Iasi the most frequent etiological agents of urinary tract infections were: E. coli - 64%, Klebsiella spp. 11% and Enterococcus spp - 5%. The resistance rate of E. coli and Klebsiella spp. was 41% and 60%, respectively to amoxicillin-clavulanic acid, 29.6% and 72.5%, respectively to third generation cephalosporins, 26% and 24%, respectively to ciprofloxacin. The most active antimicrobial agents against cephalosporins resistant strains of E. coli and Klebsiella spp were carbapenems (susceptibility rate 99% and 94%, respectively) and colimycin (susceptibility rate 89% and 83%, respectively). PMID- 19295024 TI - [Retroprospective clinical epidemiological research on leptospirosis cases hospitalised in the Infectious Diseases Clinic of Iasi]. AB - MATERIAL AND METHOD: Analytical assessment was performed for a sample of 499 patients admitted in the Infectious Diseases Clinic of Iasi, in order to highlight some clinical epidemiological and laboratory characteristics. RESULTS: During the third trimester, 45.3% of patients were hospitalized, comparatively with 7.8% during the first trimester, leading to a summer - autumn seasonality. Age groups of 31-60 years registered 57.3%; 90.2% were males, and only 13.2% agricultural workers. Clinical forms and evolution were assessed by testing a number of 10 biological and biochemical indicators. Leptospira icterohaemorrhagiae was the causal agent in 46.1% of cases, followed by L. wolffi (44.3%), L. pomona (4.2%), and l. grippotyphosa (3.4%). Average length of stay of 11-20 days (42.3%) and over 20 (21.4%) represented an indicator of unfavourable medical and socioeconomic involvements due to leptospirosis. PMID- 19295025 TI - [Anthropological study on hearing deficit]. AB - Professional training of hearing impaired persons is conditioned by the stage of psychologic development and by educational level. MATERIAL AND METHOD: The study was carried out on a series of 142 children (87 girls and 55 boys) aged 12 to 17 years with hearing deficit from a special school for deaf. RESULTS: The distribution of these children according to their parents being deaf or not, skills, constitution, intensity of voice was analyzed. The goal of this study is to demonstrate that students in these special schools have to get full support for participating in joint activities with hearing students. Such activities are favorable not only for developing mutual interests, but also for improving verbal expression in a new, pleasant, lively and instructive. PMID- 19295026 TI - [Results of etiologic diagnosis in clinical syndrome consistent with acute and chronic borreliosis]. AB - Borreliosis is a multisystem infection, which in the absence of adequate diagnosis and clinical management, may develop towards various clinical forms of chronic pathology. Due to the heterogeneity of clinical manifestations it is known under more names: erythema migrans, Lyme disease, neuroborreliosis etc. MATERIAL AND METHOD: Taking into account the present interest and the weight in pathology of syndromes consistent with the suspicion of a Borrelia spp. infection, since 2002 we applied in current practice the investigation of this etiology. There have been investigated 481 subjects, clinically suspected of Borrelia spp. infection that had historical risk of tick bite and cases of serous meningitis, after exclusion of usual etiology. Tests were performed on ELISA kits with standardised immunoreagents and recently, for result validation, on Western immunoblot kits (WB). RESULTS: Our results revealed the Borrelia etiology in 32% of cases (27.96-36.29% CI = 95%) at the screening, value expressed by the persistent positivity of the specific immunoglobulins (Ig) IgM (80.5%) and IgM+IgG (19.5%). Historic infection, represented exclusively by IgG positivity, was present in 8.6% (5.87-11.98% CI = 95%) from the cases that were negative for IgM (68%, 63.71-72.04%, CI = 95%). This weight is superposable with the results obtained in investigating a comparable sample of healthy individuals (193 subjects with 6.74% historical IgG, 3.79-10.96%, CI = 95%). Based on these results, it can be considered that ELISA procedure is useful and of reliable prognosis value for screening the Borrelia spp. etiology, the next step, taking into account the higher sensitivity of WB, being WB procedure which is useful for confirmation of ELISA positive cases and for treatment efficiency surveillance. The results prove that Borrelia spp. infections are a public health issue, which due to the diversity of clinical manifestations and diagnosis difficulties need repeated and complex laboratory investigations. PMID- 19295027 TI - [Multidisciplinary prospective epidemiological research regarding the prevalence of ankylosing spondilytis, comparatively in Romania and Iasi County, between 1990 2006]. AB - During the period of study, the prevalence of ankylosing spondylitis (AS) cases values varied between 5.65% per hundred thousand in 1994 and 3.96 in 1999. RESULTS: Are close to those obtained in the national research in 1989 (5.29% hundred thousand) and 1997 (6.34% hundred thousand). The sample of 6894 cases registered a prevalence of 28.86% in the 35-44 age groups with a decreasing trend, and 0.07% in = 85 age group. Males' prevalence was of 84.08%, comparatively with females (15.91% from total cases). Difference between residence areas had not statistic significance. In Iasi County, AS prevalence had increased values in all 17 years of study period, greater and often double that those at national level, and also greater than every county from Moldavia and one from the Western Romania. Demographical structure, variations, and those occupational, ecological, economical, educational could explain this situation. CONCLUSION: Ankylosing spondylitis is included among the major problems in order to promote community health, taking into account its clinical epidemiological characteristics. PMID- 19295028 TI - Oral rehabilitation between reality and paradigm. AB - The oral rehabilitation represents the integrative concept which re-establishes every segment of the dental-jaw system that is affected by edentation in different clinical forms, without eluding the induced complications and the influence of general status within the chosen therapy algorithm. Despite the significant development of knowledge and techniques, it is still controversial to which degree the therapy solutions of oral rehabilitation approach the therapy's ideal, and also the way in which particular aspects of the prosthetic treatment, the general status and socioeconomic criterion influence the clinical result. The implant-prosthetic therapy represents the state of art in modern treatment of edentulous patients, starting from the simplest cases and ending up with complex situations. However the problem of choosing the best treatment option remains, the patient being confused between several possibilities and numerous limits. In the best of cases, the patient understands, accepts and can afford the adequate treatment option. In other cases, choosing the therapeutically solution can be made only under the impulse of external factors. In critical situations, the patient's fear, disbelief, the restrained economic possibilities exclude the ideal solution and compromise the long-term success of the treatment. Several complex cases that were treated accordingly to the ideal option, using complex integrated algorithms, support the idea that future economic development and improved education should surpass present obstacles, the reality becoming a paradigm. PMID- 19295029 TI - [Use of laser-therapy for treatment of oral and maxillofacial hemangioma]. AB - Laser-therapy opened new perspectives for the treatment of vascular lesions of the maxillofacial regions. MATERIAL AND METHOD: The study evaluated the results of diode laser percutaneous coagulation of oral and maxillofacial hemangioma. A group of 12 patients with sonographically certified hemangioma was treated using percutaneous coagulation with a diode laser. RESULTS: The results showed good and rapid healing, without any complications. All patients presented size reduction of the lesions, a favorable esthetic appearance and no morbidity. There were no complications over a follow-up period of 3 years. The study recommends diode laser-therapy as a valuable treatment option for hemangioma in correctly selected indications. PMID- 19295030 TI - Aesthetic goals in porcelain fused to metal prosthetic appliances. AB - The ultimate goal for ceramic fused to metal restoration is the creation of a perfect aesthetic integration of the restoration with healthy tissues. MATERIAL AND METHOD: The study included a group of twenty-one patients. Sixteen of them were partial edentulous patients and five presented dental lesions or chromatic modifications of the dental tissues. The treatment plan consisted of metal ceramic crowns and bridges. The ceramic system used was Vintage Halo (Shofu). RESULTS: According to the patient's age, topography of the missing teeth or colour modification, we surveyed the aesthetic rehabilitation, creating the specific morphological elements, in order to realise an individual natural aspect with a similar optical behaviour as the dental enamel. CONCLUSIONS: The current popularity of metal-ceramic restorations is primarily related to their predictable strength and reasonable aesthetics. The main parameters that are responsible for a high-quality result are represented by: non-invasive and reversible procedures, conservative preparations, individual function offered by an individualized reconstruction, gnathology-based concepts into the occlusal rehabilitation, biocompatible materials, longevity of the reconstruction and comfort for the patient. PMID- 19295031 TI - Characterization of some extractive fractions isolated from raw Allium cepa L. bulbs. AB - Many recent studies have showed that onion (Allium cepa L.) has antioxidant effect which could be of great importance for prevention and treatment of different diseases: diabetes, cancer, cardiovascular and neurodegenerative disorders. The aim of this study was to investigate the antioxidant capacity of two extracts isolated from fresh onion bulbs: an hydroalcoholic extract (codified EC1) and an aqueous extract (codified EC2). METHODS: Antioxidant capacity was evaluated by three different assays: DPPH scavenging assay, H2O2, scavenging assay and reducing power assay. RESULTS: Both investigated extracts possess interesting antioxidant properties which are significantly influenced by their chemical composition. PMID- 19295032 TI - The study of some polyphenolic compounds from Melissa officinalis L. (Lamiaceae). AB - We proposed the analyses of polyphenolic coumpounds from the Melissa officinalis L. (Lamiaceae) leaves obtained from Medicinal Plant Collection of USAMV Cluj Napoca. MATERIAL AND METHOD: The study was performed by using spectrophotometric (I) and HPLC methods (II). RESULTS: The vegetal product contains 0.64% flavonoids expressed in rutoside and 8,962% phenyl-propane derivatives expressed in caffeic acid (I). HPLC analyses (II) were made after extraction of studied compounds from leaves with ethyl-ether, ethyl acetate and 1-buthanol. These extracts were analyzed before and after the hydrolysis of compounds. There were identified 6 polyphenolic compounds: caftaric acid, caffeic acid, p-cumaric acid, ferulic acid, luteolin and apigenin. CONCLUSION: The extracted amount of these compounds in chosen solvents depending of their polarity. PMID- 19295033 TI - Degradation of polyvinyl chloride (PVC) / hydrolyzed collagen (HC) blends active sludge test. AB - Biodegradable polymers represent a solution for the environment protection: they decrease the landfill space, by declining the petrochemical sources, and offer also an alternative solution for the recycling. MATERIALS AND METHODS: The behavior during degradation in the presence of active sludge of some polyvinyl chloride (PVC) based blends with variable content of hydrolyzed collagen (HC) has been followed. Some samples were subjected to UV irradiation, for 30 hours. The modifications induced in the environment by the polymer systems (pH variation, bacterial composition), as well as the changes of the properties of the blends (weight losses, aspect etc.) were studied. RESULTS: During the first moments of degradation in active sludge, all the samples absorbed water, behavior which favored the biodegradation. The bacteriological analysis of the sludge indicates the presence of some microbiological species. Generally, the populations of microorganisms decrease, excepting the sulphito-reducing anaerobic bacteria, the actinomycetes and other anaerobic bacteria. CONCLUSIONS: PVC/HC blends are degraded with a significant rate in active sewage sludge. More susceptible for the degradation are the UV irradiated blends. After the migration of the components with a small molecular mass in the environment, the natural polymer is degraded. The degradation effect increases with the content in the natural polymer. PMID- 19295034 TI - MIDAS intelligent platform for medical services, support for decision optimization in virtual medical communities. AB - The paper describes the implementation of a open multifunctional platform--MIDAS- for heterogeneous medical data management--support for optimization of clinical decision in virtual medical communities. The objectives of this intelligent environment are: diagnostic easier by access to heterogeneous medical data, a virtual support for medical personal in order to reduce medical errors, fast access to resources for education and improvement of medical education for physicians and students. The structure of the platform is based on a core module and a number of dedicated modules that give an important advantage as re configurable platform depending on necessities. The core module tries to be as general is possible in order to be used in the future as core model in a platform focused on dentistry cases. PMID- 19295035 TI - A psycho-verbal stimulation interface for the cognitive evoked potentials acquisition. AB - The psycho-verbal stimulation interface, created by us, represents an IT extension of an EEG / EMG device dedicated for the human brain Evoked Potentials acquisitions (EPs). METHODS: The psycho-verbal stimulation interface was created as a .NET application and was written in the C# language with respect to the OOP paradigm. It uses the TL logic to command the EPs acquisition module. The signal is send on the parallel port (LPT1) to the EPs device by a dedicated hardware interface. The software module was designed as a multi-threading application in order to perform more than one operation once. RESULTS: The stimulus sample can be customized as text, image or both and each stimulus sequence follows also a fully configurable time schema. The application can measure the patient's reaction time to each stimulus sample, which are finally centralized on a list control. The system can save both the test sequences and the EEG diagrams in digital formats; the resulted files can be stored and delivered online using the Telemes web application. CONCLUSIONS: The Telmes project implemented a secured & scalable tele-medical centers network dedicated to the telemonitoring & tele consulting services, and the psycho-verbal stimulation interface is one of the instruments devoted to these goals. The psycho-verbal stimulation interface can be used as a medical research tool for study the cognitive processes of reading, memory or learning using the endogenous visual event related potentials, as good as a instrument of training the recovery of sensitive language, which can be delivered at home by the neuro-linguist specialist as daily lists programs. The visual evoked potentials and the reaction times collected from the patients can facilitate a prognostic diagnosis of recovery the language. PMID- 19295036 TI - [Scaffolds based on collagen and chitosan for post-burn tissue engineering]. AB - Porous scaffolds based on collagen and chitosan have been obtained from mixed bio polymeric solutions and mixture freeze-drying method in the purpose of using them as materials for post-burns tissue regeneration. MATERIAL AND METHOD: Soluble collagen from bovine leather was obtained by acid-base extraction (isoelectric pH = 4.82). Two types of chitosan (CS I, M(w) = 755.900, de-acetylating degree of 79.2% and CS II, M(w) = 309.900, de-acetylating degree of 79.7%), were provided by Vascon Co., Canada. Various compositions were prepared and then structurally and morphologically characterized. In vitro degradation studies were performed in buffered collagenase or chitosan solutions, respectively, and the kinetic data were analysed. Materials effect on the tissue regeneration was tested on heat induced burns in Wistar rats by covering the damaged tissue with collagen chitosan scaffolds for a period of 28 days. Materials were changed every 7 days. At the end of the follow-up period skin tissue samples were harvested for histological investigation. RESULTS: By freeze-drying of collagen-chitosan solutions porous scaffolds were obtained with a lamellar morphology and porosity closer to chitosan than to collagen. In vitro degradation tests in simulated body fluid with collagenase revealed a decrease of the degradation rate of the collagen by mixing with chitosan. By using chitosan with lower molecular weight the degradation rate of the materials was decreased too, and the influence of the proportion of chitosan in composition diminished; stronger interactions between polymers hinder the enzyme diffusion to the following amino-acids groups: glycine - leucine (Gly-Leu), glycine - isoleucine (Gly-Ile), alanine-proline glycine/leucine (-Ala-Pro-Gly-/-Leu-). In vivo tests and histological examination revealed a differentiated repair process of the post-combustion wounds in accordance with the scaffold-type influence. CONCLUSION: Scaffolds based on collagen and chitosan are biocompatible materials with promising results for tissue regeneration of the wounds. PMID- 19295037 TI - [The measurement of nerve root tensile stress]. AB - In the majority of the cases sciatica is caused by an intervertebral disc herniation compressing the nerve root. The compression determines an increased tension in the nerve root. We believe that the magnitude of this stress offers more information about the root impairment that the disc displacement measured on MRI images. MATERIAL AND METHODS: We present in this paper an original device which allows for intraoperative root stress analysis. The device consists of a force transducer composed of an elastic element and a displacement limitation. RESULTS: The initial measurements were performed on silicon catheters and human cadavers in two different situations: 1. free nerve root; 2. blocked nerve root. Next step was intraoperative nerve roots tensile stress recording during lumbar disc surgery. Different force values were obtained for the same displacement (3 mm perpendicular on nervous root): F1 = 0.21 N; F2 = 0.78 N. Considering experimental values, the tensile stress inside of a nervous root is determined by using specific mechanical calculations presented in this paper. This is a simple and useful device for rapid intraoperative recording of nerve root mechanical stress. PMID- 19295038 TI - [Antoine Bechamp and Victor Cornil Memento for Romanian pharmacy, chemistry and medicine]. AB - Today we commemorate a century from the death of two Romanian loving scientists: Antoine Bechamp (1816- March 31 1908), pharmacist, chemist and physician, professor at the Universities of Strasbourg, Montpellier and Lille, and Victor Cornil (1837- April 14 1908), physician, histopathologist and bacteriologist, professor of the University of Medicine in Paris, co-author with Victor Babes on the first ever Bacteriological Treaty. PMID- 19295039 TI - Assessing need and distribution of neonatal intensive care beds in Mississippi. PMID- 19295040 TI - Access to medications and intermediate markers of health outcomes of a clinic population of patients with diabetes. AB - This study is a correlational analysis of the efficacy of a Medication Assistance Program (MAP) on intermediate physiologic markers of health outcomes of patients with diabetes, specifically blood pressure, HBA1c, and lipid levels. METHODS: The sample included 72 patients enrolled in a MAP and 854 patients not enrolled in a MAP and with a diagnosis of diabetes whose key elements of care are tracked in a registry. RESULTS. For established clinic patients, when compared to MAP patients, the MAP group had a small but statistically significant lower mean systolic blood pressure and higher HbA1c. Comparing MAP patients new to the program and those that were established in the program, there were no significant differences on blood pressure, HbA1c, and lipid level. CONCLUSIONS: Provision of medication alone is certainly necessary, but may not be sufficient, to address health outcomes for impoverished populations. PMID- 19295041 TI - Clueless. PMID- 19295042 TI - Address of the President, 140th Annual Session, May 30, 2008, Biloxi Beau Rivage. "Bringing it home". PMID- 19295043 TI - Has our MSMA been hijacked? PMID- 19295044 TI - SGR, GDP, and CRAP. PMID- 19295045 TI - Prevalence of obesity and overweight among children and youth in Mississippi: current trends in weight status. AB - The purpose of this research was to estimate the 2007 prevalence of obesity and overweight among children and youth and to compare these data with previous prevalence data to determine current trends in weight status in Mississippi. Measured height and weight data were collected on a weighted sample of 3,837 public school students in grades K-12. Prevalence estimates of obesity and overweight, as indicated by Body Mass Index, were calculated. Analyses were made by gender, race, and grade level for 2007 data, and these data were compared to the 2005 findings. In 2007, 23.5% of the students were obese. The prevalence was highest among elementary school students, males and nonwhite students. The prevalence of obesity in 2007 was lower than in 2005, and the decline occurred primarily among middle and high school students. While the 2007 prevalence was still high, the decline suggests that recent obesity initiatives may be having a positive impact. PMID- 19295046 TI - A pain in the neck. PMID- 19295047 TI - How to get off the Medicare merry-go-round. PMID- 19295048 TI - Never. PMID- 19295049 TI - Confabulation nation. PMID- 19295050 TI - Effects of Helicobacter infection on research: the case for eradication of Helicobacter from rodent research colonies. AB - Infection of mouse colonies with Helicobacter spp. has become an increasing concern for the research community. Although Helicobacter infection may cause clinical disease, investigators may be unaware that their laboratory mice are infected because the pathology of Helicobacter species is host-dependent and may not be recognized clinically. The effects of Helicobacter infections are not limited to the gastrointestinal system and can affect reproduction, the development of cancers in gastrointestinal organs and remote organs such as the breast, responses to vaccines, and other areas of research. The data we present in this review show clearly that unintentional Helicobacter infection has the potential to significantly interfere with the reliability of research studies based on murine models. Therefore, frequent screening of rodent research colonies for Helicobacter spp. and the eradication of these pathogens should be key goals of the research community. PMID- 19295051 TI - Alopecia: possible causes and treatments, particularly in captive nonhuman primates. AB - Alopecia (hair loss) occurs in some nonhuman primates housed in captivity and is of concern to colony managers and veterinarians. Here we review the characteristics, potential causes, and treatments for this condition. Although we focus on nonhuman primates, relevant research on other mammalian species is discussed also, due to the relative paucity of studies on alopecia in the primate literature. We first discuss the cycle of hair growth and explain how this cycle can be disrupted to produce alopecia. Numerous factors may be related to hair loss and range from naturally occurring processes (for example, seasonality, aging) to various biologic dysfunctions, including vitamin and mineral imbalances, endocrine disorders, immunologic diseases, and genetic mutations. We also address bacterial and fungal infections, infestation by parasites, and atopic dermatitis as possible causes of alopecia. Finally, we examine the role of psychogenic factors, such as stress. Depending on the presumed cause of the hair loss, various treatment strategies can be pursued. Alopecia in nonhuman primates is a multifaceted disorder with many potential sources. For this reason, appropriate testing for various disease conditions should be completed before alopecia is considered to be related to stress. PMID- 19295052 TI - Kinetics of transmission, infectivity, and genome stability of two novel mouse norovirus isolates in breeding mice. AB - Murine noroviruses are a recently discovered group of viruses found within mouse research colonies in many animal facilities worldwide. In this study, we used 2 novel mouse norovirus (MNV) wildtype isolates to examine the kinetics of transmission and tissue distribution in breeding units of NOD.CB17-Prkdc(scid)/J and backcrossed NOD.CB17-Prkdc(scid)/J x NOD/ShiLtJ (N1) mice. Viral shedding in feces and dissemination to tissues of infected offspring mice were monitored by RT-PCR over a 6-wk period postpartum. Histologic sections of tissues from mice exposed to MNV were examined for lesions and their sera monitored for the presence of antibodies to MNV. Viruses shed in feces of parental and offspring mice were compared for sequence homology of the Orf2 gene. Studies showed that the wildtype viruses MNV5 and MNV6 behaved differently in terms of the kinetics of transmission and distribution to tissues of offspring mice. For MNV5, virus transmission from parents to offspring was not seen before 3 wk after birth, and neither isolate was transmitted between cages of infected and control mice. Susceptibility to infection was statistically different between the 2 mouse strains used in the study. Both immunodeficient NOD.CB17-Prkdc(scid)/J mice and NOD. CB17-Prkdc(scid)/J x NOD/ShiLtJ offspring capable of mounting an immune response shed virus in their feces throughout the 6-wk study period, but no gross or histologic lesions were present in infected tissues. Progeny viruses isolated from the feces of infected offspring showed numerous mutations in the Orf2 gene for MNV5 but not MNV6. These results confirm previous studies demonstrating that the biology of MNV in mice varies substantially with each virus isolate and mouse strain infected. PMID- 19295053 TI - Use of low-molecular-weight heparin to decrease mortality in mice after intracardiac injection of tumor cells. AB - Intracardiac injection of human tumor cells into anesthetized nude mice is an established model of bone metastasis. However, intracardiac injection of some human tumor cell lines cause acute neurologic signs and high mortality, making some potentially relevant tumor cell lines unusable for investigation. We showed that intracardiac injection of tumor cells can induce a hypercoagulable state leading to platelet consumption and thromboemboli formation and that pretreatment with intravenous injection of low-molecular-weight heparin (LMWH; enoxaparin) blocks this state. In addition, intravenous injection of enoxaparin before intracardiac injection with 2 different small-cell lung carcinoma lines, H1975 and H2126, dramatically decreased mouse mortality while still generating bone metastases. Therefore, reduction of mortality by pretreatment with LMWH increases the types of cells that can be studied in this metastasis model and decreases the number of animals used. PMID- 19295054 TI - Effects of 4-vinylcyclohexene diepoxide on peripubertal and adult Sprague-Dawley rats: ovarian, clinical, and pathologic outcomes. AB - Young rats treated daily with intraperitoneal 4-vinylcyclohexene diepoxide (VCD) undergo selective destruction of primordial follicles, resulting in gradual ovarian failure resembling the menopausal transition in women. To determine whether VCD has similar effects on ovaries of older rats, adult and peripubertal Sprague-Dawley rats were injected intraperitoneally daily for 30 d with vehicle or VCD at 40 or 80 mg/kg. Body weight, food intake, complete blood counts, and markers of liver injury and renal function were measured during VCD treatment. Complete gross necropsy and microscopic observations were performed on day 31, and ovarian follicles were counted. At 80 mg/kg, VCD destroyed primordial and primary follicles to a similar extent in both adult and peripubertal animals, although adult rats likely started with fewer follicles and therefore approached follicle depletion. Treatment with VCD did not affect body weight, but food intake was reduced in both adult and peripubertal rats treated with 80 mg/kg VCD. Adult rats treated with 80 mg/kg VCD had neutrophilia and increased BUN and creatinine; in addition, 4 of these rats were euthanized on days 25 or 26 due to peritonitis. VCD treatment did not increase alanine aminotransferase levels, a marker of liver injury, although the 80-mg/kg dose increased liver weights. In conclusion, VCD effectively destroys small preantral follicles in adult Sprague Dawley rats, making them a suitable model of the menopausal transition of women. However, because adult rats were more sensitive to the irritant properties of VCD, the use of a lower dose should be considered. PMID- 19295055 TI - Evaluation of buprenorphine in a postoperative pain model in rats. AB - We evaluated the commonly prescribed analgesic buprenorphine in a postoperative pain model in rats, assessing acute postoperative pain relief, rebound hyperalgesia, and the long-term effects of postoperative opioid treatment on subsequent opioid exposure. Rats received surgery (paw incision under isoflurane anesthesia), sham surgery (anesthesia only), or neither and were treated postoperatively with 1 of several doses of subcutaneous buprenorphine. Pain sensitivity to noxious and nonnoxious mechanical stimuli at the site of injury (primary pain) was assessed at 1, 4, 24, and 72 h after surgery. Pain sensitivity at a site distal to the injury (secondary pain) was assessed at 24 and 72 h after surgery. Rats were tested for their sensitivity to the analgesic and locomotor effects of morphine 9 to 10 d after surgery. Buprenorphine at 0.05 mg/kg SC was determined to be the most effective; this dose induced isoalgesia during the acute postoperative period and the longest period of pain relief, and it did not induce long-term changes in opioid sensitivity in 2 functional measures of the opioid system. A lower dose of buprenorphine (0.01 mg/kg SC) did not meet the criterion for isoalgesia, and a higher dose (0.1 mg/kg SC) was less effective in pain relief at later recovery periods and induced a long-lasting opioid tolerance, indicating greater neural adaptations. These results support the use of 0.05 mg/kg SC buprenorphine as the upper dose limit for effective treatment of postoperative pain in rats and suggest that higher doses produce long-term effects on opioid sensitivity. PMID- 19295056 TI - Three-dimensional time-of-flight magnetic resonance angiography of intracranial vessels in a canine model of ischemic stroke with permanent occlusion of the middle cerebral artery. AB - The purpose of this study was to evaluate the potential efficacy of 3-dimensional time-of-flight magnetic resonance angiography (TOF-MRA) to validate a canine ischemic stroke model. Ischemic stroke was induced through permanent middle cerebral artery occlusion (MCAO) in 5 healthy Beagle dogs. T2-turbo spin echo images and TOF-MRA were obtained with a 1.5-T magnetic resonance system before and 3 and 10 d after MCAO. In 3 dogs, angiograms of the brain obtained at 3 d after MCAO showed complete occlusion of the MCA; in addition, T2 hyperintensities were present unilaterally in the striatocapsular and cerebral cortex lesions. Partial occlusion of the proximal part of the MCA was identified in the 2 remaining dogs, with T2 hyperintensities present only in the striatocapsular lesions. The occluded sites were confirmed at necropsy. The results of this study demonstrate the potential of TOF-MRA to provide a detailed description of intracranial arteries and aid in the evaluation of flow impairment in a canine MCAO model. PMID- 19295057 TI - Refinement of canine pancreatitis model: inducing pancreatitis by using endoscopic retrograde cholangiopancreatography. AB - The causes and treatments of pancreatitis have been studied in diverse species, but the canine pancreatitis model has been used most often due to its similarities to the condition in humans. Although pancreatitis in dogs can be induced readily by numerous methods, managing these dogs can be difficult because they often develop severe abdominal pain, vomiting, inappetance, and lethargy. In an effort to study pancreatitis, we performed a pilot study to determine whether an endoscopic pancreatic procedure would be possible in a dog and whether, through various manipulations, a new method of inducing pancreatitis could be developed. The model uses endoscopic retrograde cholangiopancreatography (ERCP), a common procedure in human gastroenterology that has been associated with postprocedural pancreatitis. Although all 8 dogs used in developing the ERCP model had both biochemical and histologic changes consistent with pancreatitis, 7 of the 8 dogs remained free of classic clinical signs of the disease. This method is presented as a refinement of a canine model and presents an alternative method of inducing pancreatitis, with decreased risk of developing associated clinical signs. PMID- 19295058 TI - Extreme susceptibility of African naked mole rats (Heterocephalus glaber) to experimental infection with herpes simplex virus type 1. AB - Herpes simplex virus type 1 (HSV1) is widely used as a gene delivery vector in a variety of laboratory animals. In a recent study, a thymidine-kinase-inactive (replication-conditional) HSV1 used as a delivery vector was lethal in naked mole rats, whereas mice infected with the identical virus showed no adverse effects. This result prompted us to undertake a controlled comparative histologic study of the effect of HSV1 infection on naked mole rats and mice. Replication-competent and replication-conditional HSV1 caused widespread inflammation and necrosis in multiple organ systems of naked mole rats but not mice; naked mole rats infected with replication-defective virus showed no adverse effects. We conclude that the lethality of HSV1 for naked mole rats is likely the result of overwhelming infection, possibly in part due to this species' natural lack of proinflammatory neuropeptides at the initial site of infection. PMID- 19295059 TI - Bacillus thuringiensis (Bt) transgenic crop: an environment friendly insect-pest management strategy. AB - Introduction of DDT (dichloro-diphenyl-trichloroethane) and following move towards indiscriminate use of synthetic chemical insecticides led to the contamination of water and food sources, poisoning of non-target beneficial insects and development of insect-pests resistant to the chemical insecticides. Increased public concems about the adverse environmental effects of indiscriminate use of chemical insecticides prompted search of altemative methods for insect-pest control. One of the promising alternatives has been the use of biological control agents. There is well-documented history of safe application of Bt (B. thuringiensis, a gram positive soil bacterium) as effective biopesticides and a number of reports of expression of delta-endotoxin gene(s) in crop plants are available. Only a few insecticidal sprays are required on Bt transgenic crops, which not only save cost and time, but also reduce health risks. Insects exhibit remarkable ability to develop resistance to different insecticidal compounds, which raises concern about the unsystematic use of Bt transgenic technology also. Though resistance to Bt products among insect species under field conditions has been rare, laboratory studies show that insects are capable of developing high levels of resistance to one ormore Cry proteins. Now it is generally agreed that 'high-dose/refuge strategy' is the most promising and practical approach to prolong the effectiveness of Bt toxins. Although manybiosafety concerns, ethical and moral issues exist, area under Bt transgenic crops is rapidly increasing and they are cultivated on more than 32 million hectares world over Even after reservation of European Union (EU) for acceptance of geneticaly modified (GM) crops, 6 out of 25 countries have already adopted Bt crops and many otherindustrial countries will adopt Bt transgenic crops in near future. While the modem biotechnology has been recognized to have a great potential for the promotion of human well-being, adoption of biosafety protocol is necessary to protect human health and environment from the possible adverse effects of the products of genetic engineering. The debate between proponents and opponents of GM technology has created major obstacles in hamessing benefits of the technology It has now become clear that transgenics willbe accepted by the public only when doubts related with general risks and environmental safety are adequately dispelled. Thus, there is need to organize public awareness and present the benefits of Bt transgenic crops to improve social attitude for their rational deployment. In this review, an attempt has been made to discuss social and environmental safety issues of Bt transgenic crops. PMID- 19295060 TI - Evaluation of specific biological heat potential of oily wastewater in an autothermal thermophilic aerobic treatment system. AB - This study focuses on the specific biological heat potential (h(b)) of oil and grease wastewaterin an autothermal thermophilic aerobic treatment (ATAT) system. A novel experimental device was applied to evaluate h(b) by using heat balance model under steady state. In the study the treatment system was daily fed with realistic and artificial wastewater at 11250 and 17420 mg COD l(-1), respectively. The wastewater was rich in oil and grease at 1220 and 600 mg l(-1), respectively. The sludge retention time (SRT) was controlled at 5 days. The results showed that the average values of h(b) were 3.7 and 3.1 kcal g(-1) COD removed and the true growth yield (Y0) were 0.10 and 0.13 mg MLSS mg(-1) COD for realistic and artificial wastewater respectively. These two systems could maintain reactoroperating temperatures at 43 degrees C and 48 degrees C, respectively. The COD removal efficiency was as high as 90 to 97%. The oil and grease reduction was 68 to 72%. The high organic matter removal capacity and low sludge yield of ATAT process have been demonstrated. PMID- 19295061 TI - Ontogenetic changes in foliar features and psoralen content of Psoralea corylifolia Linn. exposed to SO2 stress. AB - Field-grown Psoralea corylifolia plants were exposed to 0.5 ppm and 1.0 ppm concentrations of sulphur dioxide gas and sampled for observation at the pre flowering, flowering and post-flowering stages of plant ontogeny. One ppm SO2 concentration caused a significant decline in leaf number and leaf area per plant, total leaf dry weight, and the size and amount of midrib vasculature. The density and size of stomata decreased and many stomata were damaged. Interestingly new epidermis developed oversome of the damaged leaf stomata, thus showing a unique defence strategy against SO2 stress through dedifferentiation of the epidermal cells. Decline in the concentrations of leafchlorophylls and carotenoids in treated plants were up to 20% and 29% respectively. Stomatal conductance, intercellular CO2 content and net photosynthetic rate lowered byover 52%, 20% and 35%, respectively under the SO2 stress. Concentration of psoralen, a basic linear furanocoumarin known for its use in the treatment of dermal diseases, was highest (5.32%) in seeds and lowest (0.28%) in roots. It was heavily reduced in SO2 treated plants, the maximum decline occurring in seeds (86.70%) and leaves (56.27%). In the roots and shoots of the treated plants, it was low in pre-flowering stage, compared with the control, but showed a recovery during the post-flowering phase of plant growth. PMID- 19295062 TI - Larvicidal activity of an indigenous plant, Centratherum anthelminticum. AB - Crude extracts of fruits and leaves of Centratherum anthelminticum in different solvents were tested for larvicidal activity against Anopheles stephensi, the vector of malaria. The petroleum ether crude extract of both fruits and leaves exhibited significant larvicidal activity against III instar larvae with LC50 values of 162.60 ppm and 522.94 ppm, respectively after 24 hr. The petroleum ether extract of fruit was 11.66, 2.15 and 1.32 times more toxic than that of leaf extract after 24, 48 and 72 hr, respectively at LC90 level. However at LC50 level the corresponding values were 3.22, 1.83 and 1.19, respectively. The petroleum ether extract of C. anthelminticum fruits is a promising source for the control of Anopheles larvae. PMID- 19295063 TI - Impact of de-silting of pond on the growth and survival of catla (Catla catla) spawn. AB - Experiments were conducted in non-desilted and de-silted earthen nursery ponds for a period of fifteen days to evaluate the growth and survival of Catla spawn. The fry attained an average weight of 42.03 +/- 0.36 and 74.36 +/- 72 mg, with a survival rate of 23.44 +/- 0. 31 and 50.91 +/- 0.21% in non-desilted and de silted ponds respectively Mentioned length data also indicate that growth and survival of Catla spawn was significantly higher in de-silted ponds. PMID- 19295064 TI - Effect of modifying land cover and long-term agricultural practices on the soil characteristics in native forest-land. AB - Natural forestland soils in the high land mountain ecosystems on the eastern Black sea region of Turkey are being seriously degraded and destructed due to intensive agricultural practices. In this study we examined four soil profiles selected from four sites in each of three adjacent land use types which are native forest, pasture and cultivated fields with corn and hazelnut to compare the soil physical, chemical and morphological properties modified after natural forestland transformation into cultivated land. Disturbed and undisturbed soil samples were collected from four sites. The effects of agricultural practices on soil properties taken from each three adjacent land use types were most clearly detected in the past 50 years with the land use change. Land use change and subsequent tillage practices resulted in significant decreases in organic matter, total porosity, total nitrogen and reduced soil aggregates stability. However, contents of available P were improved by application of phosphorous fertilizers in cultivated system. There was also a significant change in bulk density among cultivated, pasture and natural forest soils. Depending upon the increase in bulk density and disruption of pores by cultivation, total porosity decreased accordingly. The data show that long term continuous cultivation of the natural forest soils resulted in changes in physical and chemical characteristics of soils. PMID- 19295065 TI - Biomonitoring of metal deposition by using moss transplant method through Hypnum cupressiforme (Hedw.) in Mussoorie. AB - Metals Cu, Zn, Cd and Pb were surveyed at 14 sampling sites by using moss Hypnum cupressiforme through active monitoring technique. Samples were transplanted in all four directions of Mussoorie city and were harvested after exposure of four months (representing each season) to analyze metal precipitation and its trend at different sites during 2005. Bioaccumulation ability for metals was evaluated seasonally exhibiting maximum in summer followed by winter and minimum in rainy season. However, at some places Cu shows highly significant values in rainy season in comparison to winter. In case of Zn and Pb significantly different (p < or = 0.05) values were observed between summerand rainy season. Baseline concentration of Cu, Pb and Zn was significantly different at 5% in comparison to other transplant sites. Result indicates Dhanaulti as most polluted location might be due to higher tourist activity and vehicular load, whereas, same was found low at Chamba might be due to place was free from pollution sources or away from in proximity to road and have low human interference. The present study allows us to determine the extent of the area affected by metal precipitation load in different rural and urban areas and abundance of metals in order of Zn > Pb > Cu > Cd. PMID- 19295066 TI - Studies on the changes in lipid peroxidation and antioxidants in drought stress induced cowpea (Vigna unguiculata L.) varieties. AB - Cowpea (Vigna unguiculata L.) is one of the major vegetable crops cultivated in tropical conditions. Two varieties of cowpea, i.e., Vigna unguiculata L.cv Kanakamony and cv Pusakomal were selected for the present study. The changes in lipid peroxidation, ascorbic acid content and activities of enzymatic antioxidants associated with drought stress were determined. A high level of lipid peroxidation was observed in test plants subjected to water stress. The Pusakomal leaves with severe stress showed 2.7 fold increase in TBARS compared to control and 1.2 fold to that of Kanakamony The drought tolerant variety showed significant increase in the activities of peroxidase and catalase on exposure to drought stress. Kanakamony leaves in severe stress possess 1.09 times increase in peroxidase and 1.8 times increase in catalase activity than Pusakomal. The concentration of ascorbic acid in test plants depleted with increased drought stress in both varieties. While the leaves of tolerant variety in severe stress showed 1.13 times higher ascorbic acid content than Pusakomal. Among the two varieties, Kanakamony possess the best antioxidant system to tolerate drought stress. This could limit cellular damage caused by active oxygen species, during waterdeficit. The variety Pusakomal was relatively poor in these adaptations. PMID- 19295067 TI - Water quality and zooplankton composition in a receiving pond of the stormwater runoff from an urban catchment. AB - Six storm periods were monitored from November 2002 to September 2005 at two stations of a receiving pond of the stormwater runoff from a small urban catchment of the city of Santa Fe, Argentina. Weekly samples were taken before and after rain events under different conditions of temperature, pluvial precipitation, and duration of the previous dry period. A sampling station was established at the outlet of the catchment (S1) and another one near the outlet of the receiving pond (S2). Both stations differed significantly in their dissolved oxygen (DO) concentration, temperature, transparency, and zooplankton composition. The concentrations of nutrients and BOD5 values indicated permanently eutrophic condition at both stations. After rainstorms, the concentrations of lead, zinc and suspended solids showed a marked increase. The zooplankton composition at S1 was characterized by the abundance of protozoans (Dexiostoma campylum (Stokes) Didinium nasutum Muller, Plagyopila cf nasuta, and Bdelloidea rotifers (Philodina sp and Rotaria sp), while Monogononta rotifers and small cladocerans were dominant at S2. The most abundant species were the rotifers Platyias quadricornis (Ehrenberg), Mytilina ventralis (Ehrenberg) and Lepadella ovalis (Muller), and the cladoceran Chydorus pubescens Sars. PMID- 19295068 TI - Water quality and pollution status of Chambal river in National Chambal Sanctuary, Madhya Pradesh. AB - The physico-chemical characteristics of Chambal river water in National Chambal sanctuary (Madhya Pradesh) have been studied. The stretch of Chambal river contained in the National Chambal sanctuary (located at 25 degrees 23'-26 degrees 52'N, 76 degrees 28'-79 degrees 15'E) is extending up to 600 km downstream from Kota (Rajasthan) to the confluence of the Chambal with Yamuna river (Etawah). The river flow in Madhya Pradesh spans up to approximately 400 km. Three sampling stations viz., Station A--near Palighat, district Sheopurkalan, Station B--near Rajghat, district Morena and Station C--near Baraighat, district Bhind were established for the collection of water samples during April, 2003 to March, 2004. The water quality parameters namely transparency (12.12-110 cm), colour (transparent-very turbid), turbidity (1-178 TNU), electrical conductivity (145.60 884 microS cm(-1)), total dissolved solids (260-500 mgl(-1)), pH (7.60-9.33), dissolved oxygen (4.86-14.59 mgl(-1)), free carbon dioxide (0-16.5 mgl(-1)), total alkalinity (70-290 mgl(-1)), total hardness (42-140 mgl(-1)), chloride (15.62-80.94 mgl(-1)), nitrate (0.008-0.025 mgl(-1)), nitrite (0.002-0.022 mgl( 1)), sulphate (3.50-45 mgl(-1)), phosphate (0.004-0.050 mgl(-1)), silicate (2.80 13.80 mgl(-1)), biochemical oxygen demand (0.60-5.67 mgl(-1)), chemical oxygen demand (2.40-26.80 mgl(-1)), ammonia (nil-0.56 mgl(-1)), sodium (14.30-54.40 mgl( 1)) and potassium (2.10 mgl(-1)-6.30 mgl(-1)) reflects on the pristine nature of the river in National Chambal sanctuary. On the basis of various parameters studied, Chambal river in this stretch can be placed under the category of oligosaprobic. The water quality analysis, indicated that the riverwater in the sanctuary area is pollution free and can serve as a good habitat for many aquatic animals including endangered species. PMID- 19295069 TI - Ostracods as indicators of pollution in the lakes of Mysore. AB - Ostracods are small crustaceans found in aquatic habitats and the present paper deals with the role of water quality index on their population diversity and seasonal fluctuations in the four lakes of Mysore city. The present investigation was carried out from July 2004 to June 2005. The study revealed highest water quality index and population density of ostracods during summer and least during winter. Dalvoi lake recorded higher water quality index (125.04), population density (60 l(-1)) but lower species diversity (2 species) of ostracods, whereas Kamana lake recorded lowest water quality index (63.49), population density (40 l(-1)) and highest species diversity (6 species) of ostracods. Increase in the water quality index indicates increase in the pollution load. As water quality index (WQI) increases, population density of ostracods increases but species diversity decreases. PMID- 19295070 TI - Ecological study of rotifers from Kor river, Fars, Iran. AB - In the present study, water samples from 3 stations of Kor river, Iran were analyzed for seasonal fluctuation of physico-chemical parameters, i.e. air and water temperature, dissolved oxygen, BOD, COD, total nitrogen, total hardness, pH, alkalinity, acidity, total dissolved solid and water current. Effect of physico-chemical parameters on surface water density of rotifers was also studied. During one year study 13 species of rotifers belonging to 2 classes and 3 orders, 6 families and 10 genera were recognized. Maximum population density belongs to Lecane luna and the minimum population density was observed in Keratella valga and Monostyla lunaris. An inverse relationship between the population density of rotifera and the water current as wall as a direct relationship between the water temperature and the population density of rotifers was observed in all sampling stations and throughout the study period. PMID- 19295071 TI - Impact of copper on the oxidative metabolism of the fry of common carp, Cyprinus carpio (Linn.) at different pH. AB - In order to evaluate the impact of copper on the energetics of a fish, the levels of glucose, glycogen, pyruvate and lactate, the rate of tissue oxygen consumption and the activities of glycogen phosphorylase, isocitrate dehydrogenase (ICDH), succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) were estimated in the whole body of the fry of Cyprinus carpio immediately after 1, 7, 15 and 30 days on exposure to a sublethal concentration of copper 0.08 mgl(-1) at pH 7.5 (normal), 6.0 (weak acidic) and 9.0 (weak alkaline). Aprogressive increase in glucose level and glycogen phosphorylase activity with the corresponding decrease in glycogen level over the time of exposure at pH 7.5 indicated glycogenolysis. Increase in the rate of oxygen consumption, pyruvate level and ICDH and SDH activities at days 1 and 7 (day 1 > 7) followed by their decrease at days 15 and 30 (day 15 < 30) at pH 7.5 indicated an initial elevation in the energetics of the fish fry with a gradual suppression of it on prolonged exposure. During this period the animal might have relied more on energetically less efficient glycolysis as evident by the progressive increase in the level of lactate and LDH activity. The degree of glycogenolysis was relatively more at pH 6.5 than at pH 7.5. At that pH, a progressive decrease in glucose level with an increase in the pyruvate and lactate levels and in LDH activity and a decrease in the rate of oxygen consumption and ICDH and SDH activities revealed greater reliance of the fish on anaerobic glycolysis than on oxidative metabolism. At pH 9.0 also the fish fry initially exhibited glycogenolysis, but gradually it came to normal on day 30 (day 1 > 7 > 15 > 30). Decrease in the glucose level, increase in pyruvate level, rate of oxygen consumption, and ICDH and SDH activities at all the days of exposure suggested an elevation in oxidative metabolism, but it also came to normal on prolonged exposure. Even the lactate level and LDH activity initially increased but gradually reached to normal on day 30. These results indicated that copper suppresses the energetics of the fish fry at pH 6.0, elevates at pH 9.0 relative to the changes at pH 7.5 suggesting that the toxicity of copper is dependent on pH of the water. PMID- 19295072 TI - Seasonal variations in physico-chemical characteristics of water, sediment and soil texture in arid zone mangroves of Kachchh-Gujarat. AB - The present study was carried out to determine the physicochemical characteristics of water and sediment and the textural aspects of sediments in western mangroves of Kachchh-Gujarat, west coast of India, for a period of two years during 1999-2000. Surface water and sediment temperatures varied from 17 degrees C to 37 degrees C and from 18.4 degrees C to 37 degrees C respectively. Tidal amplitude varied from 0.03 m to 3.78 m. Salinity varied from 34.0 to 44 per thousand and the pH in water and sediment ranged between 7.0 and 8.9 and 6.29 and 8.45 respectively. Variation in dissolved oxygen content was from 3.42 to 5.85 ml l(-1). Concentrations of nutrients viz. nitrate (0.23 to 7.26 microM), nitrite (0.04 to 0.87 microM), phosphate (0.13 to 3.12 microM) and reactive silicate (4.23 to 19.02 microM) also varied independently Total organic carbon varied from 0.29% to 2.56% and the total inorganic phosphorus ranged between 0.12 mg g(-1) and 1.97 mg g(-1). Total nitrogen varied from 0.02 mg g(-1) to 1.95 mg g(-1). Sediment textures ranges in terms of % of sand, clay and silt were: 0.26-19.2; 7.6-47 and 47-87.4 respectively in all the 3 stations. The nature of soil texture is characterized by the abundance of silty loam, silty clay and silty clay loam. PMID- 19295073 TI - Substituted phenols as pollutants that affect membrane fluidity. AB - In a toxicological context, the cellular effects of a variety of molecular compounds interacting with membranes may be understood in terms of their ability to affect and modulate lipid-membrane physical properties and even slight changes in membrane fluidity may cause aberrant function and pathological processes. Different model systems (mice splenocytes and liposomes) have been used in modelling studies of the physical effects on lipid bilayers underlying the action of membrane active phenolic compounds, considered by EPA (Environmental Protection Agency) as priority pollutants (phenol; 2-chlorophenol; 2,4 dichlorophenol; 2,4,6-trichlorophenol; pentachlorophenol; 2-nitrophenol; 2,4 dinitrophenol; 2-methyl-4,6-dinitrophenol). Membrane fluidity was assessed by fluorescence steady-state anisotropy of a fluorescent probe 1,6-diphenil-1,3,5 hexatriene (DPH). The substituted phenols increased the fluidity of cells and liposome membranes in a concentration dependent manner and the nitro substituted phenols were the most efficient perturbing the biophysical properties of the membrane. A good parallelism has been established between the results obtained with cell models and artificial liposome model systems, implying that liposomes are useful alternative systems in membrane modification studies and can be conveniently used in order to evaluate the potential toxic effect of phenol derivatives that are common environmental pollutants. PMID- 19295074 TI - Histopathological manifestations in kidney of Clarias batrachus induced by experimental Procamallanus infection. AB - Kidney of Clarias batrachus infected with Procamallanus showed varying degrees of histopathological alterations on 15, 30, 45 and 60 days post-infection. The infected kidney showed variable sized glomeruli, cloudy swelling in tubules, vacuolar/atrophic degeneration, fibrosis, mild degenerative changes in distal convoluted tubules, enlarged Bowmen's capsule, necrotic changes as well as increased granulation and hyperplasia in proximal convoluted tubules after 15 days. After 30 days of infection, the changes were rupture of Bowmen's capsule wall, degenerative changes, edema, necrosis, pyknosis, karyorrhexis and karyolysis in proximal and distal convoluted tubules, fibrosis, cloudy swelling and inflammatory lymphocytes, proliferation and shrinkage in glomeruli, and vacuolization in proximal convoluted tubules as well as cloudy swelling. After 45 days, the infected kidney showed cloudy swelling in glomeruli as well as variation in their size, infiltration of RBCs in intralobular vein and necrosis in proximal convoluted tubules, cloudy swelling in interstitium, vacuolization in the epithelial lining cells, necrosis in haemopoietic tissue and inflammatory edema. After 60 days post-infection, the changes were rupture of intralobular vein, cloudy swelling, necrosis in few proximal convoluted tubules, atrophy and shrinkage in glomeruli, distinct inflammatory edema, pyknosis, karyorrhexis and karyolysis, aggregation of lymphocytes and dilation in blood vessels. PMID- 19295075 TI - Process of rock phosphate solubilization by Aspergillus sp PS 104 in soil amended medium. AB - Aspergillus sp PS 104, a soil isolate had excellent potential to solubilize rock phosphate in vitro. The process was influenced by the presence of various concentrations of local loess (red soil). The simultaneous occurrence, in our experiment, of high levels of solubilized phosphate and synthesized citric acid, together with the lowest reached pH values, confirmed the role of citric acid in the phosphate solubilization mechanism. When the soil was present, phosphate release was better correlated than citrate synthesis with H+ concentration. Changes in soluble phosphate concentration did not follow a sigmoid pattern. The ability of organism to release phosphatase was also studied. An interesting relationship was observed between the two processes of phosphate mobilization: citric acid synthesis and phosphatase production. PMID- 19295076 TI - Studies on phytoplankton diversity in response to abiotic factors in Veeranam lake in the Cuddalore district of Tamil Nadu. AB - The phytoplankton samples were collected from the Veeranam lake for a period of one year from March-2005 to February-2006. The physico-chemical parameters of water such as air-temperature, water temperature, pH, salinity, dissolved oxygen, electrical conductivity and total dissolved solids were observed and their ranges were: 30.1-36.5 degrees C, 29.0-34.4 degrees C, 7.9-8.4, 1.2-2.5 mgl(-1), 7.6-9.2 microS and 2.5-5.2 mgl(-1). Totally 160 species of phytoplankton belonging to different taxonomic groups were identified. Among these 74 species to belong to Bacillariophyceae, 43 species to Chlorophyceae, 38 species to Cyanophyceae and 5 species to Euglenophyceae. The phytoplankton density was high (1705 cells l(-1)) during the summer season and low (760 cells l(-1)) during the winter season. Bacillariophyceae formed the dominant group. Species diversity index (H'), species richness (SR) and species evenness (J') were calculated. PMID- 19295077 TI - Biomanagement of sago-sludge using an earthworm, Lampito mauritii. AB - In the present study vermicomposting was carried out at three different concentrations of sago-sludge namely 50, 75 and 100% with an indigenous earthworm, Lampito mauritii for a period of 50 days. The fecundity of earthworm Lampito mauritii was found to be high in 50%. At the end of 50 days composting period. There were about 12 cocoons, 5 juveniles and 2 nonclitellates appeared at 50% sago-sludge concentration. The microbial analysis showed that after 40 days of composting their population stabilized and further increase in composting period did not increase their population size. Chemical analysis of Lampito mauritii worked substrates showed there is a step wise increase of nitrogen and phosphorus. The fold increase of phosphorus and nitrogen were found to be high for sago-sludge undergoing vermicomposting than the control. From the initial value of 1.8, 1.4 and 0.5 mg kg(-1) total nitrogen increased in a stepwise manner and reached a value of 5.8, 3.9 and 2.3 mg kg(-1), respectively for vermicomposting at 50, 75 and 100%. During composting the organic carbon decreased from its initial value of 56, 74 and 107 mg kg(-1) to 15, 25 and 58 mg kg(-1) for vermicomposting and 34, 45 and 72 mg kg(-1) for 50, 70 and 100% control, respectively. The results indicate that 50% and 75% concentration of sludge mixed with bedding material was ideal for the vermicomposting. PMID- 19295078 TI - Bio-waste from tobacco industry as tailored organic fertilizer for improving yields and nutritional values of tomato crop. AB - The dried tobacco waste was mixed in proportions of 1%, 2% and 3% in soil and filled in earthen pots of 6 kg capacity. Three replicate pots for each soil concentration were used. Tomato saplings of cultivar Bezosheetal were transplanted age of 20 days (signifying stage before flowering), 25 days (signifying stage at the start of flowering) and 45 day (signifying just at the onset of fruiting). The morphological parameters like plant height, number of leaf, flower plant(-1), number of fruits and yield plant(-1) were recorded. Nutrient parameters like moisture, total soluble solids (TSS), acidity vitamin C, reducing sugar proteins, pectin and lycopene were analyzed in tomato fruits. Experiments revealed that the yield of tomato fruit and critical nutritional parameters showed significant increase. For higher yield (183 g plant(-1)) of tomato, the best option is the use of 3% tobacco waste after 45 days of transplant. Alternatively, use of 2% tobacco waste with 45 days of transplant provides a higher quality tomato. The yield obtained here would be significantly higher than control but not as high as with 3% tobacco waste. PMID- 19295079 TI - Evaluation of physical and chemical parameters of river Kaveri, Tiruchirappalli, Tamil Nadu, India. AB - The physical, chemical and total heterotrophic bacteriological parameters (THBP) bacterial population were measured at three sampling stations namely Upper Anicut (S1), Chinthamani road bridge (S2) and Grand Anicut (S3) for a period of three month January to March 2007 in the river Kaveri, Tiruchirappalli. The pH ranged from 6.7 to 7.8, the level of solids and total hardness was high in station S2 when compared to other stations. Station 2 registered a minimum level of DO (3.6 mg l(-1)), and the BOD and COD varied from 3.47 to 4.27, 20.67 to 41.47 mg l(-1) respectively. THBP colony count was found to be high in S2 (8.16 x 10(2) CFU ml( 1)) and a lowest level in S1 (2.64 x 10(2)) and S3 (1.80 x 10(2)). It was observed in the present study that in reservoir (anicut) stations factors like pH, TDS, TSS, Hardness and THBP were at lower level than for the water at Chinthamani road bridge (S2), probably due to the mixing of municipal sewage at this point. PMID- 19295080 TI - Effect on histological and sperm kinetics in DBP exposed Wistar rats. AB - Di-n-butyl phthalate (DBP) is a ubiquitous environmental pollutant, extensively used as a softener for polyvinyl chloride resins. A study was conducted to evaluate its effect on reproductive function of Wistar rats. DBP was given orally at a dose of 500, 1000 and 1500 mg kg(-1) body weight for 7 days. Evaluating histological and fertility parameters assessed reproductive function. Significant reduction in seminiferous tubule diameter, Leydig cell nuclear diameter (except at dose 500 mg), number of primary spermatocytes, secondary spermatocytes and spermatids were observed. Caudal sperm density and viability reduced significantly. Decrease in serum testosterone was also observed. Evidence indicates that DBP exposure causes dose dependent testicular toxicity and has the potential to induce adverse effect. PMID- 19295081 TI - Biosorption of Cr (VI) ions from electroplating industrial effluent using immobilized Aspergillus niger biomass. AB - A fungus, Aspergillus niger was chemically treated with 0.1 M H2SO4 and 0.1 N NaOH to form biosorbent and it was immobilized in calcium alginate beads. The biosorption capacity of immobilized biosorbents for Cr (VI) was found to depend on pH, contact time, biosorbent dose and initial concentration of Cr (VI). The maximum uptake of Cr (VI) was 92.5, 95.9 and 98.4 mg respectively at a pH of 1.5 and with an increase in pH up to 10.5 the metal uptake decreased gradually up to 38.75, 50.19 and 65.28 mg respectively for acid treated, untreated and base treated fungal biosorbents. Increase in biosorbent dose up to 1 g of biomass and contact time up to 60 min resulted in an increase in biosorption from 19.6, 15.6 and 26.1 mg at a biosorbent dose of 0.1 g 100 ml(-1) to 92.45, 95.7 and 98.52 mg at a biosorbent dose of 1.0 g 100 ml(-1) and then further increase in adsorbent dose and contact time did not resulted in more Cr (VI) adsorption by per unit weight of biosorbent. The value of Kad (adsorption rate constant) revealed the pseudo-first order nature of biosorption. The percentage metal uptake by the biosorbent was found to decrease upto 62.33, 52.67 and 83.5 percent respectively for acid treated, untreated and base treated fungal biosorbents at the 300 mgl( 1) Cr (VI) ion concentration. The resulted data was found to fit well in Langmuir model of adsorption isotherm with a high value of correlation coefficient. The value of Qmax, b (Langmuir constants), R(L) (separation factor) and delta G (Gibb's free energy) revealed the favourable nature of adsorption. The biosorbed metal was eluted from the biosorbent by using 0.1 M H2SO4 as elutant. Immobilized biosorbent can be reused for five consecutive biosorption/desorption cycles without apparent loss of efficiency after its reconditioning. The biosorbent was found to perform well in the electroplating industrial effluent. PMID- 19295082 TI - Applicability of leachates originating from solid-waste landfills for irrigation in landfill restoration projects. AB - Since, landfill areas are still the most widely used solid waste disposal method across the world, leachate generated from landfills should be given importance. Leachate of landfills exerts environmental risks mostly on surface and groundwater with its high pollutant content, which may cause unbearable water quality. This leads to the obligation for decontamination and remediation program to be taken into progress for the landfill area. Among a number of alternatives to cope with leachate, one is to employ the technology of phytoremediation. The main objective of this study was to determine the N accumulation ratios and the effects of landfill leachate in diluted proportions of chosen ratios (as 1/1, 1/2, 1/4, 0), on the growth and development of Cynodon dactylon, Stenotaphrum secundatum, Paspalum notatum, Pennisetum clandestinum, Mentha piperita, Rosmarinus officinalis, Nerium oleander, Pelargonium peltatum and Kochia scoparia species. In order to simulate the actual conditions of the landfill, soil covering the landfill is taken and used as medium for the trials. The study showed that S. secundatum, K. scoparia and N. oleander species had an impressive survival rate of 100%, being irrigated with pure leachate, while the others' survival rates were between 0 to 35% under the same conditions. As expected, application of leachate to the plants caused an increase in the accumulation of N, in the upper parts of all plants except P. peltatum. The highest N content increase was observed at S. Secundatum set, accumulating 3.70 times higher than its control set, whereas P. clandestinum value was 3.41 times of its control set. PMID- 19295083 TI - Spectral reflectance properties of mangrove species of the Muthupettai mangrove environment, Tamil Nadu. AB - In the Muthupettai mangrove environment, spectral properties of six mangrove species viz. Avicennia marina, Aegiceras corniculatum, Excoecaria agallocha, Acanthus ilicifolius, Suaeda monoica and S. maritima was studied using Multi band Ground Truth Radiometer (Model-041). The study found that the chlorophyll concentration of different mangrove leaves varies between 0.05 and 0.36 mg g(-1), registering the minimum in S. maritima and maximum in E. agallocha. Interestingly species with higher chlorophyll concentration showed lower reflectance values alteast in the spectral bands 1 and 2. E. agallocha registered 0.36 mg g(-1) of chlorophyll while it recorded only 2.18 and 2.43% reflectance where as S. maritima recorded 3.16 and 3.27% of reflectance in bands 1 and 2. This indicates chlorophyll concentration is one of major factors responsible in determining the reflectance pattern of the pant communities. The spectral properties of mangroves were largely differed with that of the water and soil samples collected from the same locations, these results favourd the utilization of remotely sensed data for depicting various water and soil quality parameters from that of mangrove species in the mangrove environment. This study also found that the difference in reflectance of mangroves at canopy level is not only influenced by the chlorophyll content of species but also by the prevailing environmental condition and background reflectance of soil and water as well. PMID- 19295084 TI - Effect of gamma rays on different explants of callus treatment of multiple shoots in Cucumis melo cv. Bathasa. AB - A mutagenesis programme was carried out using physical mutagens (gamma rays) on Cucumis melo cv. Bathasa. In irradiated seeds the number of shoots formed in the lower doses was significantly higher than the controls. Decrease in the number of shoots and shoot bud formation was observed with higher doses in all the explants studies. There was complete lethality in the 10, 15 and 20 kR doses. In irradiated stem cultures the maximum number of shoots were observed in 2 kR. The number of shoots decreased with increasing doses of irradiation. At higher doses of 4 and 5kR, light green compact callus was formed in almost all the explants. The effect of lower doses of gamma irradiation on shoot bud formation and rooting efficiency from nodal explants cultured on MS +2.0 mgl(-1) L-glutamic acid + 0.5 mgl(-1) BAP, stimulation of shoot and root induction were studied. The number of shoots and root lengths decreased with the increasing dosage of irradiation. The irradiated callus was grown on solidified MS medium with containing 2.0 mgl(-1) BAP + 1.0 mgl(-1) IAA. There was a significant stimulation of growth in the callus at lower doses. At higher doses like 15 and 20 kRs growth was drastically reduced. PMID- 19295085 TI - Impact of sewage irrigation on speciation of nickel in soils and its accumulation in crops of industrial towns of Punjab. AB - Analysis of soil samples collected from sewage and tube well irrigated soils of Ludhiana, Amritsar Jalandhar and Mandi Gobindgarh, revealed that Diethylene triamine pentaacetic acid extractable nickel ( DTPA-Ni) was found to be higher in sewage fed soils. Sewage irrigation increased soil DTPA-Ni content by 3.04 times over the tube well irrigated soils. The content of DTPA-Ni showed decreasing trend with depth. Hydrogen concentration (pH) was negatively and significantly correlated with DTPA-Ni nickel whereas, organic carbon and total Ni show positive and significant correlation. Sequential fractionation was carried out to partition Ni in to fractions namely exchangeable and water soluble, organic bound, carbonate bound, Mn oxides bound, amorphous Fe oxides, crystalline Fe oxides bound and residual. Plant availability of these fractions is believed to decrease in the above order. Sequential fractionation indicated that every extracted fraction exhibited increase in Ni content with sewage irrigation with most prominent increases occurring in the organic and oxide fractions. The lowest amount of Ni in exchangeable and water soluble and the highest in residual pools testify that plants grown on these soils may not suffer from Ni toxicity. Though all the crops irrigated with sewage water had appreciably higher concentration of Ni as compared to the crops raised with tube-well water yet raya (Brassica juncea) and toria (Brassica campestris) accumulated higher content of heavy metals as compared to other crops, with higher content in roots than shoots. Transport index suggested that major part of taken up Ni is translocated to top parts of plant. Based on values of transport indices, different crops maybe arranged as toria > raya = maize > bajra > lady finger. As the plants take up nickel readily and there is danger of its excessive accumulation in plant organs and devaluation of the plant products. This is topical issue particularly in crops used for direct consumption. PMID- 19295086 TI - Utilization of various nitrogen, phosphorus, and selenium compounds by Cochlodinium polykrikoides. AB - The abilityof Cochlodinium polykrikoides to use various nitrogen, phosphorus, and selenium compounds as a nutrient source was examined in batch culture experiments to obtain biological information on the mechanism of C. polykrikoides bloom formation. C. polykrikoides grew using a variety of organic or inorganic nitrogen sources except for L-proline. Maximum yields of C. polykrikoides were obtained by gradually increasing ammonium from 5 to 20 microM, but the yield was inhibited by addition of more than 50 microM. Growth was observed in media containing various phosphorus sources, such as phosphate and 11 different organic compounds. Organic nitrogen and phosphorus seem to play an important role in the dominance of phytoplankton species and mass growth of C. polykrikoides. The ability to use a variety of organic nutrients may allow C. polykrikoides to grow to a high density in spite of inorganic nitrogen and phosphorus depletion. C. polykrikoides grew in the presence of selenite, selenate, and Se-(methyl) selenocysteine hydrochloride. However, growth yield was inhibited by addition of more than 500 microM selenate and 1000 microM Se-(methyl) selenocysteine hydrochloride. Therefore, much of the oil effluents seen in 2005 may have a temporary inhibitory action on C. polykrikoides growth but can be expected to have a positive effect in the long term. PMID- 19295087 TI - Triadimefon pretreatment protects newly assembled membrane system and causes up regulation of stress proteins in salinity stressed Amaranthus lividus L. during early germination. AB - Imposition of salinity stress during early germination imposes a secondary oxidative stress in 120-hr-old Amaranthus lividus seedlings (measured in terms of accumulation of reactive oxygen species, antioxidative defense system and oxidative membrane lipid and protein damages). Seeds of Amaranthus when treated with triadimefon along with NaCI salinity significantly enhanced the activities of catalase, peroxidase and superoxide dismutase, compared to untreated salinity stressed 5-day-old seedlings. Triadimefon treatment also reduced the accumulation of both the ROS (H2O2 and O2*-) in 5-day-old Amaranthus seedlings. When oxidative membrane damages were estimated for triadimefon treated and salinity stressed juvenile seedlings and compared with untreated salinity stressed seedlings, it shows a clear reversal in oxidative membrane damages induced by triadimefon under salinity stress. Triadimefon treatment significantly reduces the membrane lipid peroxidation and the loss of membrane protein thiol level in salinity stressed Amaranthus seedlings. That triadimefon treatment under salinity stress restores the membrane integrity and improves the post-germinative seedling growth could be supported by the data of membrane injury index (MII), relative leakage ratio (RLR), membrane permeability status (MPS), relative growth index (RGI) and mean tolerance index (MTI). SDS-PAGE of total extractible proteins revealed that some new proteins were synthesized in triadimefon treated and salinity stressed seedlings as compared to untreated and salinity stressed one. However the most remarkable feature is the up-regulation of some of the stress proteins in triadimefon treated and salinity stressed seedlings. So, it appears that significant extent of salinity tolerance exhibited by triadimefon pretreated Amaranthus seedlings could be related to the mitigation of oxidative damage to the newly assembled membrane system of juvenile tissues as well as synthesis and up-regulation of stress proteins that enhanced salinity tolerance. PMID- 19295088 TI - Effect of some abiotic factors on population buildup of Idioscopus clypealis (Lethierry) in western Uttar Pradesh. AB - Idioscopus clypealis (Lethierry) is commonly known as mango leaf hopper. The hopper reduces mongo production by causing non setting of flower and dropping of immature fruits. The first appearance of hoppers (0.12 and 0.16) on the branches of mango trees was recorded in February which is correlated with increase in maximum (23.89 degrees C and 24.29 degrees C) and minimum temperature (11.46 degrees C and 11.88 degrees C) and decrease in relative humidity (88.15% and 86.85%). Peak hopper population (5.88 and 6.46) was recoded in May on temperature range of(40.34 degrees C and 40.88 degrees C) and (27.22 degrees C and 27.62 degrees C) as maximum and minimum, which was very high, whereas, relative humidity was very low (55.80% and 55.09%). With the increase in maximum and minimum temperature and decrease in relative humidity hopper population raised. Thus, maximum and minimum temperature positively affected the hopper population, whereas, relative humidity had negative effect, but rainfall showed no significant effect, as it was fluctuating. PMID- 19295089 TI - Decolorization of pulp paper mill effluent by Pleurotus sajorcaju. AB - Pleurotus sajorcaju MTCC-141 procured from Microbial Type Culture Collection Centre and Gene Bank, Institute of Microbial Technology, Chandigarh has been used for color removal from paper mill effluent. The paper mill effluent amended with basal medium supports the growth of Pleurrotus sajorcaju and removes the colour. The optimum concentrations of carbon source (glucose) and nitrogen source (NH4NO3) for the maximum decolourization of paper mill effluent were found to be 1% and 0.2% respectively. During the fungal growth process, the pH of the paper mill effluent decreased from 7.94 to 4.0. PMID- 19295090 TI - Removal of colour from textile effluent using cyanobacterial biomass. AB - The cyanobacterial species were isolated from fresh water pond of Gura-vishnoiyan and were tested for their ability to decolourise the textile effluent. The reduction in colour of textile effluent with the use of dry biomass of Cylindrospermum indicum, Nostoc calcicola, Calothrix weberi was comparatively observed after 2, 4 and 6 days, but maximum colour absorption was observed after 6 days. Calothrix weberi was found the most effective species of cyanobacteria, which can remove the colour intensity upto 85 percent after 6 days of incubation. However, Nostoc calcicola and Cylindrospermum indicum showed 45 percent and 23 percent of colour reduction respectively. PMID- 19295091 TI - Hydrochemical assessment of the pollutants in groundwaters of Vrishabhavathi Valley Basin in Bangalore (India). AB - The present study aims at the assessment of groundwater quality in and around the Vrishabhavathi Valley, the erstwhile fresh water stream, today carrying huge quantities of industrial, agricultural and domestic effluents from the western part of Bangalore metropolis. Groundwater samples were collected from both bore wells and open wells along the Vrishabhavathi watershed and subjected to a comprehensive physico-chemical and bacteriological analysis. The study revealed that 57% of the samples were non-potable due to their values when compared to the BIS standards. The concentrations of nitrate and total hardness were found higher than the standards in 43.33% and 40% of the samples respectively. 50% of the samples examined, indicated bacterial contamination in the groundwater. PMID- 19295092 TI - Energy balance of a suburban vegetated area in a tropical city--Delhi (India). AB - Energy balance measurements were carried out in a suburban vegetated area of southern part of Delhi. These measurements were made on several days during winter (January, February, November, December) and summer (April, May, June) of 1998 and 1999, respectively. During the winter 1998, net radiation (Q*) reached up to 108 W/m2 around noon and during the summer 1999, Q* was as high as 423.43 W/m2. The apportionments of Q* into sensible heat (H) and latent heat (LE) fluxes were 63% and 16% respectively during the summer 1998, whereas, during the winter Q* dissipated by 40% and 60%, respectively by H and LE. Similarly, during the summer 1999, partitioning of Q* into H and LE were 52 and 24%, respectively and during the winter, apportionments of H and LE found to be 37% and 39% respectively. On an average, about 34% of Q* was dissipated as soil heat flux (G) during both the summers (1998 and 1999) and the winter 1999, whereas, during the winter 1998, G was about 14% of Q*. Diurnal variations of H and LE showed erratic nature whereas that of G was relatively smoother during both the summers and winters. The calculated Bowen ratio was 0.67 to 0.91 during winters and 2.16 to 3.84 during summers, respectively. Probably, seasonal differences are due to local/synoptic conditions, nature of sub stratum and existence of vegetation, which influence the energy partitioning. However, as there was no surplus of energy, the area acts as a heat sink. PMID- 19295093 TI - Polycyclic aromatic hydrocarbons in ambient air at Agra: distribution and toxicity assessment. AB - Polycyclic Aromatic Hydrocarbons (PAHs) are a class of organic pollutants that are commonly found in the environment, largely due to combustion or processing of hydrocarbon fuels. PAHs are considered highly toxic for human beings and several of these compounds are carcinogenic, mutagenic and teratogenic. Human exposure to PAHs occurs principally by direct inhalation, ingestion or dermal contact as a result of the widespread presence and persistence of PAHs in the urban environment. With increasing awareness that PAHs are known and suspected carcinogens, this study was undertaken to monitor PAH compounds in Total Suspended Particulate Matter (TSPM) at the industrial site--Nunhai, Agra. For this purpose, TSPM samples were collected on glass fibre filter papers (EPM 2000) using High Volume Sampler (HVS 430) at Nunhai, Agra. 16 EPA priority PAH compounds were analyzed by a Gas Chromatograph equipped with FID detector. PAHs having high molecular weight, i.e., BghiP, BbF, DbA, BaA, BaP and IP, were the most abundant with concentrations ranging between 0.026 to 0.56 ng m(-3). The calculated mean TPAH value was 0.32 ng m(-3). The probably carcinogenic and possibly carcinogenic PAH as classified by International Agency for Research on Cancer (IARC) accounted for 42% and 38% respectively of the total PAH. The health risk associated with inhalatory exposure to PAHs was assessed on the basis of Benzo(a)pyrene concentration in air and Toxic Equivalency Factor (TEF) for individual PAH. In the present study, BaP concentrations ranged from 0.005 to 0.23 ng m(-3) with a mean value of 0.04 ng m(-3). Carcinogenic potencies for DbA and BaP in PAH mixtures based on TEF concept were 45% and 39% respectively. This underlines the importance of DbA and BaPas a surrogate compound of a PAH mixture in our environment in assessing human health risk. PMID- 19295094 TI - Drinking water quality assessment of rain water harvested in ferrocement tanks in Alappuzha District, Kerala (India). AB - The study was conducted to assess the physico-chemical and bacteriological quality of stored rain water in the ferrocement tanks of Alappuzha District, Kerala (India). Representative water samples were collected on random basis from ten stations (S1 to S10) with rain water harvesting facility during the periods January 2006 and April 2006. The present study revealed that the physico-chemical characteristics of stored rain water analysed during the winter and summer seasons were within the permissible drinking water standard limits prescribed by W.H.O. Microbiological analysis showed that most of the stored rainwater samples had microbial contamination in both winter and summer seasons and the bacterial count was above the permissible standards for drinking water. Faecal coliforms were also detected in the stored rain water samples collected from eight stations during the summer season. The present study revealed that the quality of stored rain water is suitable for drinking purpose in terms of physical and chemical characteristics. However, disinfection is necessary to make the water microbially safe for drinking purposes. PMID- 19295095 TI - Fluoride contamination in ground water of Malaprabha sub basin. AB - The fluoride contamination study has been carried out in ground waters of Khanapur, Bailhongal and Saundatti taluks of Malaprabha sub basin, Belgaum district, Karnataka (India). 21 samples were collected from different locations covering shallow and deep aquifers. The samples were collected during the pre monsoon and post-monsoon season of 1999.The fluoride concentrations varied from 0.55 to 3.85 mg/L during pre-monsoon and 0.20 to 3.50 mg/L during post-monsoon seasons. Higher concentration of fluoride was observed in Ugargol village of Saundatti taluk. It was observed that about 28.60% of the Samples have fluoride concentration less than 1 mg/L, 23.80% have concentration in the range of 1.0 1.50 mg/L, 42.85% have concentration in the range of 1.50-3.0 mg/L and 4.75% have concentration more than 3.0 mg/L during the pre-monsoon. During post-monsoon, it was found that about 35% of the samples have fluoride concentration less than 1.00 mg/L, 25% have concentration in the range of 1.0-1.50 mg/L, 35% have concentration in the range of 1.50-3.0 mg/L and 5% have concentration more than 3.0 mg/L, A positive correlation of fluoride was observed with bicarbonate, alkalinity and pH during the pre-monsoon and with EC, TDS, carbonate, bicarbonate and potassium during the post-monsoon. This suggests that source of fluoride may be partially from potassium rich rocks and constituents of fertilizers leaching down to ground water. The regression equations have been developed between fluoride and other water quality parameters. PMID- 19295096 TI - Heavy metals leaching in Indian fly ash. AB - Fly ash is an industrial waste generated from thermal power plants. Fly ash constitutes 80-85% of the total ash produced. A small part of fly ash is utilised in some sectors such as construction materials, building engineering, road, back fill, agriculture, selective engineering and processing useful materials. A large part of fly ash produced is disposed of with very high environmental risk. In the present paper, laboratory leaching test has been used to determine the potential mobility of Pb, Cd, Cr, Cu, Zn, Fe, Mn and Ni in fly ash samples, collected from Chandrapura Thermal Power Plant, Jharkhand and Ramagundam Super Thermal Power Plant, Andhra Pradesh, in order to assess their leachability when these wastes are disposed of. A cascade-leaching test was used at liquid-to-solid ratio (L/S) ranging between 20 and 100. Both fly ash samples exhibited neutral reactions, as indicated by pH values <11.75 and >7.0 at L/S=10 and contact time of 10 minutes. The percentage of leached amounts found to follow the trend Zn>Fe>Mn>Cr>Pb>Cu>Ni>Cd for fly ash from Chandrapura and Fe>Zn>Cu>Mn>Cr>Ni>Pb>Cd for fly ash from Ramagundam. Effect of pH on metals released from ash surface in aqueous solution followed a predictable pattern of decreasing release with increasing pH. PMID- 19295097 TI - Assessment of hand pump waters in three tribal dominated districts of southern Rajasthan, India. AB - Udaipur, Banswara and Dungarpur districts of southern Rajasthan (India) have dominance of tribal population. In these districts besides other water resources, hand pumps are catering the need for drinking water. The present study was undertaken to assess the level of chemical and bacteriological status for comparing the water quality with the prevailing standards. 18 hand pumps were studied for selected water quality parameters such as, pH, hardness, chloride, fluoride, NO3 -N, EC, orthophosphate, TDS, TSS, BOD, total coliforms and faecal coliforms, following the standard methods. The data on chemical parameters revealed that in all 18 hand pumps the water quality was within the permissible level of WHO. However, in eight hand pumps the faecal coliforms were higher (2-6 MPN/100 mL) than the permissible limit which confirm organic contamination in these drinking water resources. On the basis of this study, suitable remedial measures for protection of water quality have been suggested. PMID- 19295098 TI - Removal of lead from aqueous effluents by adsorption on coconut shell carbon. AB - The application of adsorption for removal of heavy metals is quite popular and activated carbon is universally used as an adsorbent. However, high cost of its preparation and regeneration has led to a search for alternative sorbents, especially in the developing countries. A number of sorbents are used to remove metals by adsorption from industrial effluents, which include insoluble starch, xanthates, modified cotton and wool, tree barks, activated carbon, plant leaves and agricultural products. Therefore, as an alternative, coconut shell carbon (CSC), a low cost sorbent derived from organic waste material, was used in the present work, for removal of lead from aqueous effluents. The results of the batch sorption studies indicated that the efficiency of lead removal by coconut shell carbon is comparable to that of commercially available activated carbon. From the kinetic and equilibrium studies, the sorptive capacity of coconut shell carbon for lead was found to be 30 mg/g. Desorption and subsequent recovery of the metal from the surface of the sorbent was successfully demonstrated. Parameters affecting the sorption were evaluated. PMID- 19295099 TI - Design of enclosure of a grinding machine for the noise attenuation. AB - Since most of the equipments are usually installed in rooms or enclosed plants, the effect of an enclosure is important. Absorptive surface of the enclosure can reduce levels to the extent of 5 dB(A) which is quite significant. Effectiveness of enclosures in sound varies with the frequency of sound and the shape, size, laying pattern of sound absorbing material, which is put on the walls of the enclosure. The sound absorbing material accomplishes little shielding if there is no acoustical absorption within it. Acoustical absorbents when applied to the interior surface of the enclosure adsorb some of the energy and the noise level within the enclosure builds up only by the energy not absorbed. This paper elucidates the procedure of developing an enclosure to study its performance in attenuating sound at varying conditions. The results found encouraging and such enclosures can be developed for plant machineries by the plant personnels without getting confused by the claims and rhetoric of advertisements of big companies engaged in developing such items. PMID- 19295100 TI - Fuzzy rule based estimation of agricultural diffuse pollution concentration in streams. AB - Outflow from the agricultural fields carries diffuse pollutants like nutrients, pesticides, herbicides etc. and transports the pollutants into the nearby streams. It is a matter of serious concern for water managers and environmental researchers. The application of chemicals in the agricultural fields, and transport of these chemicals into streams are uncertain that cause complexity in reliable stream quality predictions. The chemical characteristics of applied chemical, percentage of area under the chemical application etc. are some of the main inputs that cause pollution concentration as output in streams. Each of these inputs and outputs may contain measurement errors. Fuzzy rule based model based on fuzzy sets suits to address uncertainties in inputs by incorporating overlapping membership functions for each of inputs even for limited data availability situations. In this study, the property of fuzzy sets to address the uncertainty in input-output relationship is utilized to obtain the estimate of concentrations of a herbicide, atrazine, in a stream. The data of White river basin, a part of the Mississippi river system, is used for developing the fuzzy rule based models. The performance of the developed methodology is found encouraging. PMID- 19295101 TI - Adsorption of fluoride on synthetic iron (III), zirconium(IV) and binary iron(III)-zirconium (IV) oxides: comparative assessment on pH effect and isotherm. AB - Fluoride is an accumulative poison at high dose of intake for humans and animals. In the present study, the sorption of fluoride from aqueous solution has been investigated on synthetic hydrous ferric oxide (HFO), hydrous zirconium oxide (HZO) and hydrous zirconium(IV)-iron(III) oxide (HZFO) by batch mode experiments. Both HFO and HZFO were crystalline and HZO was amorphous in nature. The parametes studied were the effect of pH and sorption equilibriums. The results showed increase in fluoride-sorption with increasing pH from nearly 2.0 to 5.0, 4.6 and 6.8 for HFO, HZO and HZFO, respectively. Analysis of temperature dependent sorption data obtained at equilibrium solution pH 6.8 (+/- 0.2) has been described by the Langmuir, Freundlich, Temkin and Redlich-Peterson isotherm model equations. The present sorption data fit, in general, found very well with the Langmuir and Redlich-Peterson models; and the data fit for HZFO and HFO found to increase, but for HZO the data found to decrease with increasing temperature. The computed thermodynamic parameters such as deltaG0, delltaH0 and deltaS0 from the Langmuir equilibrium constant (b, L/Umg) values show that the fluoride-sorption on HZFO was more spontaneous and endothermic process compared to HFO. The deltaH0 value obtained for fluoride adsorption on HZO indicates exothermic nature. PMID- 19295102 TI - 2, 4 dichlorophenol (2, 4-DCP) sorption from aqueous solution using granular activated carbon and polymeric adsorbents and studies on effect of temperature on activated carbon adsorption. AB - Adsorption equilibrium, kinetics and thermodynamics of 2,4-dichlorophenol (2,4 DCP), one of the most commonly used chlorophenol, onto bituminous coal based Filtrasorb-400 grade granular activated carbon, were studied in aqueous solution in a batch system with respect to temperature. Uptake capacity of activated carbon found to increase with temperature. Langmuir isotherm models were applied to experimental equilibrium data of 2, 4-DCP adsorption and competitive studies with respect to XAD resin were carried out. Equilibrium data fitted very well to the Langmuir equilibrium model. Adsorbent monolayer capacity 'Q0, Langmuir constant 'b' and adsorption rate constant 'k(a)' were evaluated at different temperatures for activated carbon adsorption. This data was then used to calculate the energy of activation of adsorption and also the thermodynamic parameters, namely the free energy of adsorption, deltaG0, enthalpy of adsorption, deltaH0 and the entropy of adsorption deltaS0. The obtained results showed that the monolayer capacity increases with the increase in temperatures. The obtained values of thermodynamic parameters showed that adsorption of 2,4 DCP is an endothermic process. Synthetic resin was not found efficient to adsorb 2,4 DCP compared to activated carbon. The order of adsorption efficiencies of three resins used in the study found as XAD7HP > XAD4 > XAD1180. PMID- 19295103 TI - Relevance of non-tuberculous mycobacteria in India. PMID- 19295104 TI - Psycho-social dysfunction: perceived and enacted stigma among tuberculosis patients registered under revised national tuberculosis control programme. AB - OBJECTIVE: To examine the perceived and enacted stigma experienced by TB patients and the community. METHODS: We interviewed 276 TB patients registered for treatment during January-March 2004 in government health facilities of two Tuberculosis Units of south India. Data on perceived and enacted stigma were collected after two months of starting treatment, using a semi-structured interview schedule. In addition, four Focus Group Discussions were conducted among Directly Observed Treatment (DOT) providers and community members. Narrative summaries were also taken down to collect additional qualitative information. RESULTS: Of the 276 patients, 190 (69%) were males. There was no significant difference between the genders in relation to social stigma. Perceived stigma was higher than enacted stigma in both genders and significantly higher among males (Low self esteem p < 0.05), change of behavior of community (p < 0.05), ashamed to cough in front of others (p < 0.05). CONCLUSION: Considering the social and emotional impact of the disease, it is essential to adopt support strategies to enhance acceptance and for a successful health programme. PMID- 19295105 TI - Study of relapse and failure cases of CAT I retreated with CAT II under RNTCP--an eleven year follow up. AB - OBJECTIVE: To analyse the treatment outcome of Cat I smear positive relapse and failure cases and their fate when treated with Cat II regimen under RNTCP. METHODS: All Cat I smear positive relapse and failure TB patients treated with Category II regimen from 1994 to 2005 in a chest clinic of Delhi were analysed in this retrospective study. The re-treatment outcome data for relapse and failure cases of Cat I when treated with Cat II regimen was reviewed. RESULTS: The study population included 5576 registered as Cat I sputum positive cases in Gulabi Bagh chest clinic from 1994 to 2005. A total of 190 (3.4%) failed on Cat I regimen. Further out of 4905 (87.9%) successfully treated Cat I patients, 442 (9%) presented as relapses. The treatment success rate for relapse and failure cases of Cat I when subsequently treated with Cat II regimen were 76.4% and 48.8% respectively, with a significantly higher failure rate (27.6%) among Cat I failures subsequently treated with Cat II regimen. CONCLUSION: The failure cases of Cat I subsequently treated with Cat II were observed to have a significantly lower success rates (p < 0.05) as compared to relapse cases. The need for reappraisal of Cat II re-treatment regimen for failure cases among Cat I is suggested. PMID- 19295106 TI - Screening of bulk drug samples and anti-tuberculosis products for the presence of therapeutically less active diasteriomeric (R,S) form of ethambutol dihydrochloride. AB - BACKGROUND: The present study was carried out to screen various ethambutol dihydrochloride (EB2HCI) bulk drug samples and anti-tuberculosis (anti-TB) products for the presence of less active (R,S)-EB2HCl. METHODOLOGY: Samples of pure EB2HCl were received gratis from various companies and the formulations were procured from local market, and also from a Directly Observed Treatment Short course (DOTS) centre. Some products available in the institute from Global Drug Facility were also included in the study. In total, 5 API samples and 35 formulations containing EB2HCI were investigated. These were subjected to evaluation for the presence of (R,S)-EB2HCl using a previously published differential scanning calorimetric method. The thermograms were recorded between 25 degrees C and 250 degrees C at a rate of 10 degrees C/min. RESULTS: 1 API sample and 12 formulations were found to contain (R,S)-EB2HCI up to an extent of 30-100%. One of the DOTS centres supply was also found to contain approximately 97% of the less active isomer. CONCLUSION: The presence of therapeutically inactive form of the EB2HCl from 30-100% in approximately 30% of the products in the local market is an alarming finding, which means low quality anti-TB products are in circulation. The same may be contributory to the developing resistance of the drugs against the mycobacterium. PMID- 19295107 TI - Treatment outcome of neuro tuberculosis patients put on DOTS--an observation study from the field. AB - BACKGROUND: Effectiveness of intermittent Short Course Chemotherapy for Neuro Tuberculosis has not been well studied. There are only few reported studies on this issue in the world literature under filed conditions. Neurologists all over India are reluctant to accept Directly Observed Treatment Short course for neuro tuberculosis since its introduction in India. AIM: Assessing effectiveness of Revised National TB Control Programme (RNTCP-DOTS) regimens among neuro tuberculosis patients registered under the programme. METHODS: All the neuro tuberculosis patients referred to RNTCP for treatment were included in the study. Study population included only those patients diagnosed at higher centre and referred to RNTCP during the period Jan - Dec 2002, Alappuzha District. Diagnostic Algorithm as per RNTCP guidelines was strictly followed and treatment outcome and follow-up status were taken from tuberculosis register. No pediatric age group was included in the study. RESULTS: A total of 32 cases registered for DOTS regimen were included in the study, of whom 29 completed the treatment and all were asymptomatic at the end of treatment (85%). All patients received treatment as DOTS, but only 70% received actual DOTS. All patients were given nine months intermittent regimen as per RNTCP guidelines. Five patients died during the treatment (14%). This result shows that DOTS under field programme conditions are efficient in curing Neuro Tuberculosis. CONCLUSION: Good result was obtained with intermittent short course chemotherapy under programme conditions in neurotuberculosis. PMID- 19295108 TI - Lupus vulgaris and tuberculosis verrucosa cutis (TBVC)--a clinical, pathological and epidemiological study of 71 cases. AB - BACKGROUND: The epidemiological aspects and diagnostic problems encountered in a rural set up are largely unknown. The present study on cutaneous tuberculosis encompassing clinical, pathological and epidemiological aspects was undertaken at Rajah Muthiah Medical college and Hospital (RMMC&H), a teaching hospital mainly catering to the health needs of a predominantly rural population from villages and hamlets around Chidambaram, a taluq headquarters, located in Cuddalore district ofTamil Nadu. MATERIAL AND METHODS: During the period of study, 5744 cases (82%) of pulmonary tuberculosis and 1261 (18%) of extra-pulmonary tuberculosis were encountered. Cutaneous tuberculosis accounted for 117 (1.67%) of tuberculosis cases. RESULTS: Of the seventy-one patients with cutaneous tuberculosis, 39 had lupus vulgaris and 32 with TBVC. Lupus vulgaris was more prevalent with male pre-ponderance. The maximum incidence was seen in the second decade of life. Both LV and TBVC showed a male pre-ponderance, M:F ratio being 3:2 and 2:1 in LV and TBVC respectively. A majority of patients with LV (27 cases; 69.2%) and TBVC (31 cases; 96.8%) manifested with a single site of involvement. Lower extremities were more commonly involved among patients hailing from poor economic strata who were not habituated to the use of footwear while working out doors. Over crowding was also a contributing factor. The relationships between BCG vaccination and Mantoux test with cutaneous tuberculosis, association with pulmonary tuberculosis in addition to the underlying predisposing socio-economic factors are discussed. PMID- 19295109 TI - Status report on RNTCP. PMID- 19295110 TI - Disseminated tuberculosis manifesting as chronic pancreatitis. AB - Pancreatic involvement in tuberculosis is known but uncommon. The clinical manifestation may vary from painless obstructive jaundice due to pancreatic mass (cyst or abscess) to fever of unknown origin. Here we report a case who initially presented as acute pancreatitis relapsing into chronic pancreatitis as an initial manifestation of disseminated tuberculosis. PMID- 19295111 TI - Tubercular hepatic abscess--a rare presentation. AB - Hepatic tuberculosis is one of the rare forms of extra-pulmonary tuberculosis. The focal or nodular form presenting as tuberculoma or abscess is uncommon. Hepatic tuberculosis without involvement of lungs or other organs is even rarer. We report a rare case of primary tubercular liver abscess without involvement of any other organ of body. PMID- 19295112 TI - National workshop on public private participation (PPP) for TB control in India- a brief review. PMID- 19295113 TI - Information and communication technology: applicatioin in health sector. PMID- 19295114 TI - DOTS in HIV positive patients. PMID- 19295115 TI - ECRL--a wrist extensor with multiple uses in deformity correction. AB - Extensor carpi radialis longus muscle has been used in various types of procedures for corrective hand surgery and is a favored muscle for correction of finger clawing due to ulnar nerve palsy in leprosy because its removal leaves an insignificant motor deficit and gives a linear scar at the donor site. It is usually not paralyzed in leprosy. The muscle, being phasic, is easy to re educate. The excursion of the muscle is similar to lumbrical muscles which it substitutes. Since the muscle is dorsally located, the transfer does not lose tension due to adaptive wrist flexing habit. Its tendon is usually thick enough, can be split into two and used as graft to elongate the muscle-tendon unit or for ligament reconstruction in cases of trapezio-metacarpal joint arthritis. PMID- 19295116 TI - Borderline tuberculoid leprosy--an unusual presentation: a case report. AB - A twenty-six year old lady presented herself with multiple erythmatous, scaly, well-defined, hypo-esthetic plaques over bilateral soles, face and left leg. Histopathology revealed features consistent with borderline tuberculoid leprosy. PMID- 19295117 TI - A descriptive study of MDT services in Chhattisgarh. AB - INTRODUCTION: The study was conducted to ascertain the registration status, time lag and reasons for non/delayed registration, pattern and sources of MDT, among new cases of leprosy, initially reporting to a tertiary level institution and subsequently referred after diagnosis, to Primary Care institutions. METHODOLOGY: All new cases referred from RLTRI, Raipur to three adjoining blocks between Nov. 2005 and Oct. 2006 were listed and followed at monthly interval at the PHC/CHC, Sub center and household levels, from 2 to 6 months after RLTRI visit. RESULTS: A total of 371 cases were referred, of which 326 (87.9%) could be traced. The level of PHC/ CHC registration of cases, which was 58.0% after 2 months, rose to 80.4% after 6 months of follow up. 82.9% of the registered cases collected subsequent doses of MDT from the nearest Sub center or PHC. Main reason for non/ delayed registration were delay in reporting from patient side (30.7%), non availability of MDT at PHC (24.1%), non-availability of concerned staff (7.3%) and other operational problems (38.0%). CONCLUSION: The study highlights the need to strengthen the "MDT Services" through monitoring and supervision of leprosy activities delivered by General Health Services in post integration phase. PMID- 19295118 TI - Partnership for rehabilitation: looking beyond impairments. AB - Rehabilitation of leprosy-affected persons extends beyond the physical domain of prevention and treatment of impairments. A holistic rehabilitative approach should include addressing those problems that people may have in activities and difficulties that may prevent people from fully participating in social functions, i.e. being fully accepted as integrated members of the societies and communities to which they belong. This article highlights the activities of the Partnership for the Rehabilitation Program (PFR) of the International Nepal Fellowship (INF), Pokhara, Nepal. These activities aim to prevent, reduce or alleviate problems and difficulties that leprosy-affected persons may face in being respected and being contributing members of the communities of which they are a part. PMID- 19295119 TI - Secular trends of new leprosy cases diagnosed in Brazil during 1987-2006. AB - OBJECTIVE: To analyze trends in occurrence of new leprosy cases in Brazil in the light of changes in public health policy during the period 1987-2006. MATERIAL AND METHODS: Study of secular trends in New case detection rate (NCDR), viz. for leprosy in Brazil between 1987 and 2006. A linear trend was used to analyze separately the detection rate during the two decades 1987-1996 and 1997-2006. The cut-off point was the year with the peak detection rate in 1997. To analyze the potential increase in accessibility to diagnosis and treatment, a comparison was made between the proportion of cases diagnosed within the health services and municipalities over a 20-year period, split into four 5-year intervals. RESULTS: The variation of the NCDR between 1987 and 2006 more than doubled between 1987 and 1997, decreased from 1997 to 2000, increased up to 2003 and then dropped considerably between 2004 and 2006. The number of facilities providing health services for the 5-year periods showed a marked increase-more than doubling between the second and third and between the third and fourth periods. The number and therapeutic services are indications that Brazil is perhaps making considerable progress towards eliminating leprosy as a public health problem. PMID- 19295120 TI - Deletion rate evolution and its effect on genome size and coding density. AB - Deletion rates are thought to be important factors in determining the genome size of organisms in nature. Although it is indisputable that deletions, and thus deletion rates, affect genome size, it is unclear how, or indeed if, genome size is regulated via the deletion rate. Here, we employ a mathematical model to determine the evolutionary fate of deletion rate mutants. Simulations are employed to explore the interactions between deletions, deletion rate mutants, and genome size. The results show that, in this model, the fate of deletion rate mutants will depend on the fraction of essential genomic material, on the frequency of sexual recombination, as well as on the population size of the organism. We find that there is no optimal deletion rate in any state. However, at one critical coding density, all changes in deletion rate are neutral and the rate may drift either up or down. As a consequence, the coding density of the genome is expected to fluctuate around this critical density. Characteristic differences in the impact of deletion rate mutations on prokaryote and eukaryote genomes are described. PMID- 19295122 TI - Intraflagellar transport at a glance. PMID- 19295124 TI - Laminin is required for Schwann cell morphogenesis. AB - Development of the peripheral nervous system requires radial axonal sorting by Schwann cells (SCs). To accomplish sorting, SCs must both proliferate and undergo morphogenetic changes such as process extension. Signaling studies reveal pathways that control either proliferation or morphogenesis, and laminin is essential for SC proliferation. However, it is not clear whether laminin is also required for SC morphogenesis. By using a novel time-lapse live-cell-imaging technique, we demonstrated that laminins are required for SCs to form a bipolar shape as well as for process extension. These morphological deficits are accompanied by alterations in signaling pathways. Phosphorylation of Schwannomin at serine 518 and activation of Rho GTPase Cdc42 and Rac1 were all significantly decreased in SCs lacking laminins. Inhibiting Rac1 and/or Cdc42 activities in cultured SCs attenuated laminin-induced myelination, whereas forced activation of Rac1 and/or Cdc42 in vivo improved sorting and hypomyelinating phenotypes in SCs lacking laminins. These findings indicate that laminins play a pivotal role in regulating SC cytoskeletal signaling. Coupled with previous results demonstrating that laminin is critical for SC proliferation, this work identifies laminin signaling as a central regulator coordinating the processes of proliferation and morphogenesis in radial axonal sorting. PMID- 19295123 TI - Mechanisms of biphasic insulin-granule exocytosis - roles of the cytoskeleton, small GTPases and SNARE proteins. AB - The release of insulin from pancreatic islets requires negative regulation to ensure low levels of insulin release under resting conditions, as well as positive regulation to facilitate robust responsiveness to conditions of elevated fuel or glucose. The first phase of release involves the plasma-membrane fusion of a small pool of granules, termed the readily releasable pool; these granules are already at the membrane under basal conditions, and discharge their cargo in response to nutrient and also non-nutrient secretagogues. By contrast, second phase secretion is evoked exclusively by nutrients, and involves the mobilization of intracellular granules to t-SNARE sites at the plasma membrane to enable the distal docking and fusion steps of insulin exocytosis. Nearly 40 years ago, the actin cytoskeleton was first recognized as a key mediator of biphasic insulin release, and was originally presumed to act as a barrier to block granule docking at the cell periphery. More recently, however, the discovery of cycling GTPases that are involved in F-actin reorganization in the islet beta-cell, combined with the availability of reagents that are more specific and tools with which to study the mechanisms that underlie granule movement, have contributed greatly to our understanding of the role of the cytoskeleton in regulating biphasic insulin secretion. Herein, we provide historical perspective and review recent progress that has been made towards integrating cytoskeletal reorganization and cycling of small Rho-, Rab- and Ras-family GTPases into our current models of stimulus secretion coupling and second-phase insulin release. PMID- 19295125 TI - The E3 ubiquitin ligase LNX1p80 promotes the removal of claudins from tight junctions in MDCK cells. AB - The structural continuity of tight junctions (TJs) is consistently maintained even when epithelial cells divide and move within the cellular sheet. This process is associated with dynamic remodeling of TJs by coordinated internalization and generation of claudin-based TJ strands, but the molecular mechanism behind the regulated turnover of TJs remains largely unknown. In this study, we identified the p80 isoform of the E3 ubiquitin ligase ligand of Numb protein X1 (LNX1p80) as a protein binding to claudin-1. Interestingly, the concentration of claudins in TJs was remarkably reduced when LNX1p80 was overexpressed in MDCK cells, and there was a reduction not only in the number of TJ strands but also in the amount of detergent-insoluble claudins. We also found that LNX1p80 promoted polyubiquitylation of claudins. This ubiquitylation is dependent on its RING-finger domain and is not mediated by Lys48 of ubiquitin, which is used for protein degradation by the proteasome. Furthermore, LNX1p80 was often colocalized with claudins in vesicular structures containing markers for late endosomes and lysosomes. These findings suggest that LNX1p80 is involved in the ubiquitylation, endocytosis and lysosomal degradation of claudins, and that the turnover of TJs is regulated by ubiquitylation. PMID- 19295126 TI - The extracellular-matrix protein matrilin 2 participates in peripheral nerve regeneration. AB - Matrilins are adaptor proteins of the extracellular matrix involved in the formation of both collagen-dependent and collagen-independent filamentous networks. Although their molecular structure and binding partners have been characterized, the functional roles of the four matrilin family members in vivo are still largely unknown. Here, we show that matrilin 2, expressed in pre myelinating Schwann cells during normal development, profoundly influences the behaviour of glial cells and neurons in vitro. When offered as a uniform substrate, matrilin 2 increased neurite outgrowth of dorsal root ganglia (DRG) neurons and enhanced the migration of both cell line- and embryonic DRG-derived Schwann cells. Vice versa, axonal outgrowth and cell migration were decreased in DRG cultures prepared from matrilin-2-deficient mice compared with wild-type (wt) cultures. In stripe assays, matrilin 2 alone was sufficient to guide axonal growth and, interestingly, axons favoured the combination of matrilin 2 and laminin over laminin alone. In vivo, matrilin 2 was strongly upregulated in injured peripheral nerves of adult wild-type mice and failure of protein upregulation in knockout mice resulted in delayed regrowth of regenerating axons and delayed time-course of functional recovery. Strikingly, the functional recovery 2 months after nerve injury was inferior in matrilin-2-deficient mice compared with wild-type littermates, although motoneuron survival, quality of axonal regeneration, estimated by analyses of axonal diameters and degrees of myelination, and Schwann cell proliferation were not influenced by the mutation. These results show that matrilin 2 is a permissive substrate for axonal growth and cell migration, and that it is required for successful nerve regeneration. PMID- 19295127 TI - Three-dimensional electron microscopy reveals new details of membrane systems for Ca2+ signaling in the heart. AB - In the current study, the three-dimensional (3D) topologies of dyadic clefts and associated membrane organelles were mapped in mouse ventricular myocardium using electron tomography. The morphological details and the distribution of membrane systems, including transverse tubules (T-tubules), junctional sarcoplasmic reticulum (SR) and vicinal mitochondria, were determined and presumed to be crucial for controlling cardiac Ca(2+) dynamics. The geometric complexity of T tubules that varied in diameter with frequent branching was clarified. Dyadic clefts were intricately shaped and remarkably small (average 4.39x10(5) nm(3), median 2.81x10(5) nm(3)). Although a dyadic cleft of average size could hold maximum 43 ryanodine receptor (RyR) tetramers, more than one-third of clefts were smaller than the size that is able to package as many as 15 RyR tetramers. The dyadic clefts were also adjacent to one another (average end-to-end distance to the nearest dyadic cleft, 19.9 nm) and were distributed irregularly along T tubule branches. Electron-dense structures that linked membrane organelles were frequently observed between mitochondrial outer membranes and SR or T-tubules. We, thus, propose that the topology of dyadic clefts and the neighboring cellular micro-architecture are the major determinants of the local control of Ca(2+) in the heart, including the establishment of the quantal nature of SR Ca(2+) releases (e.g. Ca(2+) sparks). PMID- 19295128 TI - The class I bHLH factors E2-2A and E2-2B regulate EMT. AB - Functional loss of the cell-cell adhesion molecule E-cadherin is an essential event for epithelial-mesenchymal transition (EMT), a process that allows cell migration during embryonic development and tumour invasion. In most carcinomas, transcriptional repression has emerged as the main mechanism responsible for E cadherin downregulation. Here, we report the identification of class I bHLH factor E2-2 (TCF4/ITF2) as a new EMT regulator. Both isoforms of E2-2 (E2-2A and E2-2B) induce a full EMT when overexpressed in MDCK cells but without affecting the tumorigenic properties of parental cells, in contrast to other EMT inducers, such as Snail1 or class I bHLH E47. E-cadherin repression mediated by E2-2 is indirect and independent of proximal E-boxes of the promoter. Knockdown studies indicate that E2-2 expression is dispensable for maintenance of the EMT driven by Snail1 and E47. Comparative gene-profiling analysis reveals that E2-2 factors induce similar, yet distinct, genetic programs to that induced by E47 in MDCK cells. These results, together with the embryonic expression pattern of Tcf4 and E2A (which encodes E12/E47), support a distinct role for E2-2 and suggest an interesting interplay between E-cadherin repressors in the regulation of physiological and pathological EMT processes. PMID- 19295129 TI - Distinct role of PLCbeta3 in VEGF-mediated directional migration and vascular sprouting. AB - Endothelial cell proliferation and migration is essential to angiogenesis. Typically, proliferation and chemotaxis of endothelial cells is driven by growth factors such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). VEGF activates phospholipases (PLCs) - specifically PLCgamma1 - that are important for tubulogenesis, differentiation and DNA synthesis. However, we show here that VEGF, specifically through VEGFR2, induces phosphorylation of two serine residues on PLCbeta3, and this was confirmed in an ex vivo embryoid body model. Knockdown of PLCbeta3 in HUVEC cells affects IP3 production, actin reorganization, migration and proliferation; whereas migration is inhibited, proliferation is enhanced. Our data suggest that enhanced proliferation is precipitated by an accelerated cell cycle, and decreased migration by an inability to activate CDC42. Given that PLCbeta3 is typically known as an effector of heterotrimeric G-proteins, our data demonstrate a unique crosstalk between the G-protein and receptor tyrosine kinase (RTK) axes and reveal a novel molecular mechanism of VEGF signaling and, thus, angiogenesis. PMID- 19295130 TI - An interaction network of the mammalian COP9 signalosome identifies Dda1 as a core subunit of multiple Cul4-based E3 ligases. AB - The COP9 signalosome (CSN) is an evolutionarily conserved macromolecular complex that interacts with cullin-RING E3 ligases (CRLs) and regulates their activity by hydrolyzing cullin-Nedd8 conjugates. The CSN sequesters inactive CRL4(Ddb2), which rapidly dissociates from the CSN upon DNA damage. Here we systematically define the protein interaction network of the mammalian CSN through mass spectrometric interrogation of the CSN subunits Csn1, Csn3, Csn4, Csn5, Csn6 and Csn7a. Notably, we identified a subset of CRL complexes that stably interact with the CSN and thus might similarly be activated by dissociation from the CSN in response to specific cues. In addition, we detected several new proteins in the CRL-CSN interactome, including Dda1, which we characterized as a chromatin associated core subunit of multiple CRL4 proteins. Cells depleted of Dda1 spontaneously accumulated double-stranded DNA breaks in a similar way to Cul4A-, Cul4B- or Wdr23-depleted cells, indicating that Dda1 interacts physically and functionally with CRL4 complexes. This analysis identifies new components of the CRL family of E3 ligases and elaborates new connections between the CRL and CSN complexes. PMID- 19295131 TI - Reprogramming to a muscle fate by fusion recapitulates differentiation. AB - Fusion of mammalian cells to form stable, non-dividing heterokaryons results in nuclear reprogramming without the exchange of genetic material. In this report, we show that reprogramming in somatic cell heterokaryons involves activation of the canonical skeletal muscle transcription factors as well as contraction excitation genes. Thus, the effect of heterokaryon formation on gene expression is to induce a recapitulation of differentiation. Heterokaryons formed with a relatively refractory cell type, the hepatocyte cell line HepG2, revealed the importance of both MyoD expression and other unidentified cytoplasmic components, neither of which are sufficient for efficient muscle gene activation, but are synergistic. We provide evidence that de-repression by transient histone deacetylase inhibition can induce MyoD expression and increase the extent and efficiency of muscle gene transcription. Taken together, the results suggest that understanding the mechanistic basis, using a combination of approaches, and taking into account cell history, will facilitate an increase in the efficiency and fidelity of conversion from one differentiated phenotype to another desired cell type. Inherent advantages of the heterokaryon system merit further investigation in the pursuit of directed cloning. PMID- 19295132 TI - Microarray labeling extension values: laboratory signatures for Affymetrix GeneChips. AB - Interlaboratory comparison of microarray data, even when using the same platform, imposes several challenges to scientists. RNA quality, RNA labeling efficiency, hybridization procedures and data-mining tools can all contribute variations in each laboratory. In Affymetrix GeneChips, about 11-20 different 25-mer oligonucleotides are used to measure the level of each transcript. Here, we report that 'labeling extension values (LEVs)', which are correlation coefficients between probe intensities and probe positions, are highly correlated with the gene expression levels (GEVs) on eukaryotic Affymetrix microarray data. By analyzing LEVs and GEVs in the publicly available 2414 cel files of 20 Affymetrix microarray types covering 13 species, we found that correlations between LEVs and GEVs only exist in eukaryotic RNAs, but not in prokaryotic ones. Surprisingly, Affymetrix results of the same specimens that were analyzed in different laboratories could be clearly differentiated only by LEVs, leading to the identification of 'laboratory signatures'. In the examined dataset, GSE10797, filtering out high-LEV genes did not compromise the discovery of biological processes that are constructed by differentially expressed genes. In conclusion, LEVs provide a new filtering parameter for microarray analysis of gene expression and it may improve the inter- and intralaboratory comparability of Affymetrix GeneChips data. PMID- 19295133 TI - Heat shock factor-1 modulates p53 activity in the transcriptional response to DNA damage. AB - Here we define an important role for heat shock factor 1 (HSF1) in the cellular response to genotoxic agents. We demonstrate for the first time that HSF1 can complex with nuclear p53 and that both proteins are co-operatively recruited to p53-responsive genes such as p21. Analysis of natural and synthetic cis elements demonstrates that HSF1 can enhance p53-mediated transcription, whilst depletion of HSF1 reduces the expression of p53-responsive transcripts. We find that HSF1 is required for optimal p21 expression and p53-mediated cell-cycle arrest in response to genotoxins while loss of HSF1 attenuates apoptosis in response to these agents. To explain these novel properties of HSF1 we show that HSF1 can complex with DNA damage kinases ATR and Chk1 to effect p53 phosphorylation in response to DNA damage. Our data reveal HSF1 as a key transcriptional regulator in response to genotoxic compounds widely used in the clinical setting, and suggest that HSF1 will contribute to the efficacy of these agents. PMID- 19295134 TI - Identification of microRNAs with regulatory potential using a matched microRNA mRNA time-course data. AB - Over the past decade, a class of small RNA molecules called microRNAs (miRNAs) has been shown to regulate gene expression at the post-transcription stage. While early work focused on the identification of miRNAs using a combination of experimental and computational techniques, subsequent studies have focused on identification of miRNA-target mRNA pairs as each miRNA can have hundreds of mRNA targets. The experimental validation of some miRNAs as oncogenic has provided further motivation for research in this area. In this article we propose an odds ratio (OR) statistic for identification of regulatory miRNAs. It is based on integrative analysis of matched miRNA and mRNA time-course microarray data. The OR-statistic was used for (i) identification of miRNAs with regulatory potential, (ii) identification of miRNA-target mRNA pairs and (iii) identification of time lags between changes in miRNA expression and those of its target mRNAs. We applied the OR-statistic to a cancer data set and identified a small set of miRNAs that were negatively correlated to mRNAs. A literature survey revealed that some of the miRNAs that were predicted to be regulatory, were indeed oncogenic or tumor suppressors. Finally, some of the predicted miRNA targets have been shown to be experimentally valid. PMID- 19295135 TI - The RRM domain in GW182 proteins contributes to miRNA-mediated gene silencing. AB - Proteins of the GW182 family interact with Argonaute proteins and are required for miRNA-mediated gene silencing. These proteins contain two structural domains, an ubiquitin-associated (UBA) domain and an RNA recognition motif (RRM), embedded in regions predicted to be unstructured. The structure of the RRM of Drosophila melanogaster GW182 reveals that this domain adopts an RRM fold, with an additional C-terminal alpha-helix. The helix lies on the beta-sheet surface, generally used by these domains to bind RNA. This, together with the absence of aromatic residues in the conserved RNP1 and RNP2 motifs, and the lack of general affinity for RNA, suggests that the GW182 RRM does not bind RNA. The domain may rather engage in protein interactions through an unusual hydrophobic cleft exposed on the opposite face of the beta-sheet. We further show that the GW182 RRM is dispensable for P-body localization and for interaction of GW182 with Argonaute-1 and miRNAs. Nevertheless, its deletion impairs the silencing activity of GW182 in a miRNA target-specific manner, indicating that this domain contributes to silencing. The conservation of structural and surface residues suggests that the RRM domain adopts a similar fold with a related function in insect and vertebrate GW182 family members. PMID- 19295137 TI - In utero exposure to methyl isocyanate in the Bhopal gas disaster: evidence of persisting hyperactivation of immune system two decades later. PMID- 19295136 TI - Current tools for the identification of miRNA genes and their targets. AB - The discovery of microRNAs (miRNAs), almost 10 years ago, changed dramatically our perspective on eukaryotic gene expression regulation. However, the broad and important functions of these regulators are only now becoming apparent. The expansion of our catalogue of miRNA genes and the identification of the genes they regulate owe much to the development of sophisticated computational tools that have helped either to focus or interpret experimental assays. In this article, we review the methods for miRNA gene finding and target identification that have been proposed in the last few years. We identify some problems that current approaches have not yet been able to overcome and we offer some perspectives on the next generation of computational methods. PMID- 19295139 TI - We've got NERVE: a call to arms for neuroscience education. PMID- 19295140 TI - Glial modulation of pain: a step beyond. PMID- 19295141 TI - Characterization of the decision network for wing expansion in Drosophila using targeted expression of the TRPM8 channel. AB - After emergence, adult flies and other insects select a suitable perch and expand their wings. Wing expansion is governed by the hormone bursicon and can be delayed under adverse environmental conditions. How environmental factors delay bursicon release and alter perch selection and expansion behaviors has not been investigated in detail. Here we provide evidence that in Drosophila the motor programs underlying perch selection and wing expansion have different environmental dependencies. Using physical manipulations, we demonstrate that the decision to perch is based primarily on environmental valuations and is incrementally delayed under conditions of increasing perturbation and confinement. In contrast, the all-or-none motor patterns underlying wing expansion are relatively invariant in length regardless of environmental conditions. Using a novel technique for targeted activation of neurons, we show that the highly stereotyped wing expansion motor patterns can be initiated by stimulation of N(CCAP), a small network of central neurons that regulates the release of bursicon. Activation of this network using the cold-sensitive rat TRPM8 channel is sufficient to trigger all essential behavioral and somatic processes required for wing expansion. The delay of wing expansion under adverse circumstances thus couples an environmentally sensitive decision network to a command-like network that initiates a fixed action pattern. Because N(CCAP) mediates environmentally insensitive ecdysis-related behaviors in Drosophila development before adult emergence, the study of wing expansion promises insights not only into how networks mediate behavioral choices, but also into how decision networks develop. PMID- 19295142 TI - Epithelial-derived brain-derived neurotrophic factor is required for gustatory neuron targeting during a critical developmental period. AB - Brain-derived neurotrophic factor (BDNF) is expressed in epithelial targets of gustatory neurons (i.e., fungiform papillae) before their innervation, and BDNF overexpression in nontaste regions of the tongue misdirects gustatory axons to these sites, suggesting that BDNF is necessary for gustatory axons to locate and innervate their correct targets during development. To test this hypothesis, we examined the targeting of taste neurons in BDNF-null mice (bdnf(-/-)). Analysis of bdnf(-/-) mice using a combination of DiI labeling and electron microscopy revealed that taste regions were not innervated by gustatory axons. Instead, branching was increased and many nontaste regions were innervated. The increased branching by gustatory axons in these animals was facilitated by neurotrophin 4 (NT4), because branching was virtually eliminated in bdnf(-/-)/nt4(-/-) mice. No abnormalities in gustatory innervation patterns and targeting were observed in nt4(-/-) mice. Conditional removal of BDNF selectively in epithelial cells disrupted targeting at the tongue tip, where gene recombination removed bdnf by embryonic day 13.5 (E13.5). However, innervation patterns were normal in the midregion and caudal portions of the tongue, where gene recombination did not occur until E14.5. These findings demonstrate that BDNF derived from gustatory epithelia is required for gustatory axons to correctly locate and innervate fungiform papillae. In addition, they show that BDNF-mediated targeting is restricted to a critical period of development, on or before E13.5. PMID- 19295143 TI - Dynamic changes in presynaptic and axonal transport proteins combined with striatal neuroinflammation precede dopaminergic neuronal loss in a rat model of AAV alpha-synucleinopathy. AB - Little is known about key pathological events preceding overt neuronal degeneration in Parkinson's disease (PD) and alpha-synucleinopathy. Recombinant adeno-associated virus 2-mediated delivery of mutant (A53T) human alpha-synuclein into the substantia nigra (SN) under a neuron-specific synapsin promoter resulted in protracted neurodegeneration with significant dopaminergic (DA) neuron loss by 17 weeks. As early as 4 weeks, there was an increase in a dopamine metabolite, DOPAC and histologically, DA axons in the striatum were dystrophic with degenerative bulbs. Before neuronal loss, significant changes were identified in levels of proteins relevant to synaptic transmission and axonal transport in the striatum and the SN. For example, striatal levels of rabphilin 3A and syntaxin were reduced. Levels of anterograde transport motor proteins (KIF1A, KIF1B, KIF2A, and KIF3A) were decreased in the striatum, whereas retrograde motor proteins (dynein, dynamitin, and dynactin1) were increased. In contrast to reduced levels in the striatum, KIF1A and KIF2A levels were elevated in the SN. There were dramatic changes in cytoskeletal protein levels, with actin levels increased and alpha-/gamma-tubulin levels reduced. In addition to these alterations, a neuroinflammatory response was observed at 8 weeks in the striatum, but not in the SN, demonstrated by increased levels of Iba-1, activated microglia and increased levels of proinflammatory cytokines, including IL-1beta, IFN-gamma and TNF-alpha. These results demonstrate that changes in proteins relevant to synaptic transmission and axonal transport coupled with neuroinflammation, precede alpha-synuclein-mediated neuronal death. These findings can provide ideas for antecedent biomarkers and presymptomatic interventions in PD. PMID- 19295144 TI - Rapid synaptic depression explains nonlinear modulation of spectro-temporal tuning in primary auditory cortex by natural stimuli. AB - In this study, we explored ways to account more accurately for responses of neurons in primary auditory cortex (A1) to natural sounds. The auditory cortex has evolved to extract behaviorally relevant information from complex natural sounds, but most of our understanding of its function is derived from experiments using simple synthetic stimuli. Previous neurophysiological studies have found that existing models, such as the linear spectro-temporal receptive field (STRF), fail to capture the entire functional relationship between natural stimuli and neural responses. To study this problem, we compared STRFs for A1 neurons estimated using a natural stimulus, continuous speech, with STRFs estimated using synthetic ripple noise. For about one-third of the neurons, we found significant differences between STRFs, usually in the temporal dynamics of inhibition and/or overall gain. This shift in tuning resulted primarily from differences in the coarse temporal structure of the speech and noise stimuli. Using simulations, we found that the stimulus dependence of spectro-temporal tuning can be explained by a model in which synaptic inputs to A1 neurons are susceptible to rapid nonlinear depression. This dynamic reshaping of spectro-temporal tuning suggests that synaptic depression may enable efficient encoding of natural auditory stimuli. PMID- 19295145 TI - Segregated populations of hippocampal principal CA1 neurons mediating conditioning and extinction of contextual fear. AB - Learning processes mediating conditioning and extinction of contextual fear require activation of several key signaling pathways in the hippocampus. Principal hippocampal CA1 neurons respond to fear conditioning by a coordinated activation of multiple protein kinases and immediate early genes, such as cFos, enabling rapid and lasting consolidation of contextual fear memory. The extracellular signal-regulated kinase (Erk) additionally acts as a central mediator of fear extinction. It is not known however, whether these molecular events take place in overlapping or nonoverlapping neuronal populations. By using mouse models of conditioning and extinction of fear, we set out to determine the time course of cFos and Erk activity, their cellular overlap, and regulation by afferent cholinergic input from the medial septum. Analyses of cFos(+) and pErk(+) cells by immunofluorescence revealed predominant nuclear activation of either protein during conditioning and extinction of fear, respectively. Transgenic cFos-LacZ mice were further used to label in vivo Fos(+) hippocampal cells during conditioning followed by pErk immunostaining after extinction. The results showed that these signaling molecules were activated in segregated populations of hippocampal principal neurons. Furthermore, immunotoxin-induced lesions of medial septal neurons, providing cholinergic input into the hippocampus, selectively abolished Erk activation and extinction of fear without affecting cFos responses and conditioning. These results demonstrate that extinction mechanisms based on Erk signaling involve a specific population of CA1 principal neurons distinctively regulated by afferent cholinergic input from the medial septum. PMID- 19295146 TI - Environmental enrichment restores memory functioning in mice with impaired IL-1 signaling via reinstatement of long-term potentiation and spine size enlargement. AB - Environmental enrichment (EE) was found to facilitate memory functioning and neural plasticity in normal and neurologically impaired animals. However, the ability of this manipulation to rescue memory and its biological substrate in animals with specific genetically based deficits in these functions has not been extensively studied. In the present study, we investigated the effects of EE in two mouse models of impaired memory functioning and plasticity. Previous research demonstrated that mice with a deletion of the receptor for the cytokine interleukin-1 (IL-1rKO), and mice with CNS-specific transgenic over-expression of the IL-1 receptor antagonist (IL-1raTG) display impaired hippocampal memory and long-term potentiation (LTP). We report here a corrective effect of EE on spatial and contextual memory in IL-1rKO and IL-1raTG mice and reveal two mechanisms for this beneficial effect: Concomitantly with their disturbed memory functioning, LTP in IL-1rKO mice that were raised in a regular environment is impaired, and their dendritic spine size is reduced. Both of these impairments were corrected by environmental enrichment. No deficiencies in neurogenesis or hippocampal BDNF and vascular endothelial growth factor secretion were found in IL-1rKO mice that were raised in a regular environment, and both of these variables were increased to a similar degree in enriched IL-1rKO and wild-type mice. These findings suggest that exposure to an enriched environment may be beneficial for individuals with impaired learning and memory related to genetic impairments of IL-1 signaling (and possibly other genetic causes), by reversing impairments in dentate gyrus LTP and spine size and by promoting neurogenesis and trophic factors secretion. PMID- 19295147 TI - Reverse signaling by glycosylphosphatidylinositol-linked Manduca ephrin requires a SRC family kinase to restrict neuronal migration in vivo. AB - Reverse signaling via glycosylphosphatidylinositol (GPI)-linked Ephrins may help control cell proliferation and outgrowth within the nervous system, but the mechanisms underlying this process remain poorly understood. In the embryonic enteric nervous system (ENS) of the moth Manduca sexta, migratory neurons forming the enteric plexus (EP cells) express a single Ephrin ligand (GPI-linked MsEphrin), whereas adjacent midline cells that are inhibitory to migration express the cognate receptor (MsEph). Knocking down MsEph receptor expression in cultured embryos with antisense morpholino oligonucleotides allowed the EP cells to cross the midline inappropriately, consistent with the model that reverse signaling via MsEphrin mediates a repulsive response in the ENS. Src family kinases have been implicated in reverse signaling by type-A Ephrins in other contexts, and MsEphrin colocalizes with activated forms of endogenous Src in the leading processes of the EP cells. Pharmacological inhibition of Src within the developing ENS induced aberrant midline crossovers, similar to the effect of blocking MsEphrin reverse signaling. Hyperstimulating MsEphrin reverse signaling with MsEph-Fc fusion proteins induced the rapid activation of endogenous Src specifically within the EP cells, as assayed by Western blots of single embryonic gut explants and by whole-mount immunostaining of cultured embryos. In longer cultures, treatment with MsEph-Fc caused a global inhibition of EP cell migration and outgrowth, an effect that was prevented by inhibiting Src activation. These results support the model that MsEphrin reverse signaling induces the Src dependent retraction of EP cell processes away from the enteric midline, thereby helping to confine the neurons to their appropriate pathways. PMID- 19295148 TI - NKCC1-dependent GABAergic excitation drives synaptic network maturation during early hippocampal development. AB - A high intracellular chloride concentration in immature neurons leads to a depolarizing action of GABA that is thought to shape the developing neuronal network. We show that GABA-triggered depolarization and Ca2+ transients were attenuated in mice deficient for the Na-K-2Cl cotransporter NKCC1. Correlated Ca2+ transients and giant depolarizing potentials (GDPs) were drastically reduced and the maturation of the glutamatergic and GABAergic transmission in CA1 delayed. Brain morphology, synaptic density, and expression levels of certain developmental marker genes were unchanged. The expression of lynx1, a protein known to dampen network activity, was decreased. In mice deficient for the neuronal Cl(-)/HCO(3)(-) exchanger AE3, GDPs were also diminished. These data show that NKCC1-mediated Cl(-) accumulation contributes to GABAergic excitation and network activity during early postnatal development and thus facilitates the maturation of excitatory and inhibitory synapses. PMID- 19295149 TI - Phase encoding in the Mauthner system: implications in left-right sound source discrimination. AB - The paired teleost Mauthner (M)-cells and their associated network serve as an excellent system to study the biophysical basis of decision making. In teleosts, an abrupt sound evokes an M-spike, triggering a C-start escape that is usually directed away from a sound source. The response latency is minimized by electrical synapses between auditory afferents and the M-cell lateral dendrite. Here, we demonstrate that the electrical synapses also mediate phase encoding. Ramped sound pressure waves (150-250 Hz) evoked electrotonic postsynaptic potentials in the M-cell locked to two diametrically opposed phase angles that were frequency dependent but intensity independent. Phase encoding was also evident at the behavioral level underwater, because the stimuli evoked directional C-starts with an onset that was phase locked to the sound wave. In interneurons inhibitory to the M-cell, these same stimuli also evoked phase locked electrotonic postsynaptic potentials and action potentials. The resulting chemical and electrical (i.e., field effect) inhibitions functioned tonically and phasically, respectively. Phase encoding could be important in underwater sound source localization, which is thought to require a neural computation involving a phase comparison between the pressure and the directional particle motion components of sound. This computation may be implemented by an interplay between phase-dependent afferent excitation and feedforward inhibition that activates the appropriate M-cell and directs the C-start away from the sound source. PMID- 19295150 TI - Inflammation triggers synaptic alteration and degeneration in experimental autoimmune encephalomyelitis. AB - Neurodegeneration is the irremediable pathological event occurring during chronic inflammatory diseases of the CNS. Here we show that, in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis, inflammation is capable in enhancing glutamate transmission in the striatum and in promoting synaptic degeneration and dendritic spine loss. These alterations occur early in the disease course, are independent of demyelination, and are strongly associated with massive release of tumor necrosis factor-alpha from activated microglia. CNS invasion by myelin-specific blood-borne immune cells is the triggering event, and the downregulation of the early gene Arc/Arg3.1, leading to the abnormal expression and phosphorylation of AMPA receptors, represents a culminating step in this cascade of neurodegenerative events. Accordingly, EAE-induced synaptopathy subsided during pharmacological blockade of AMPA receptors. Our data establish a link between neuroinflammation and synaptic degeneration and calls for early neuroprotective therapies in chronic inflammatory diseases of the CNS. PMID- 19295151 TI - Amyloid beta-induced neuronal hyperexcitability triggers progressive epilepsy. AB - Alzheimer's disease is associated with an increased risk of unprovoked seizures. However, the underlying mechanisms of seizure induction remain elusive. Here, we performed video-EEG recordings in mice carrying mutant human APPswe and PS1dE9 genes (APdE9 mice) and their wild-type littermates to determine the prevalence of unprovoked seizures. In two recording episodes at the onset of amyloid beta (Abeta) pathogenesis (3 and 4.5 months of age), at least one unprovoked seizure was detected in 65% of APdE9 mice, of which 46% had multiple seizures and 38% had a generalized seizure. None of the wild-type mice had seizures. In a subset of APdE9 mice, seizure phenotype was associated with a loss of calbindin-D28k immunoreactivity in dentate granular cells and ectopic expression of neuropeptide Y in mossy fibers. In APdE9 mice, persistently decreased resting membrane potential in neocortical layer 2/3 pyramidal cells and dentate granule cells underpinned increased network excitability as identified by patch-clamp electrophysiology. At stimulus strengths evoking single-component EPSPs in wild type littermates, APdE9 mice exhibited decreased action potential threshold and burst firing of pyramidal cells. Bath application (1 h) of Abeta1-42 or Abeta25 35 (proto-)fibrils but not oligomers induced significant membrane depolarization of pyramidal cells and increased the activity of excitatory cell populations as measured by extracellular field recordings in the juvenile rodent brain, confirming the pathogenic significance of bath-applied Abeta (proto-)fibrils. Overall, these data identify fibrillar Abeta as a pathogenic entity powerfully altering neuronal membrane properties such that hyperexcitability of pyramidal cells culminates in epileptiform activity. PMID- 19295153 TI - A specific role of the human hippocampus in recall of temporal sequences. AB - There is a growing interest in how temporal order of episodic memories is represented within the medial temporal lobe (MTL). Animal studies suggest that the hippocampal formation (HF) is critical for retrieving the temporal order of past experiences. However, human imaging studies that have tested recency discrimination between pairs of previously encoded items have generally failed to report HF activation. We hypothesized that recalling a naturalistic sequence of past events would be particularly sensitive to HF function, attributable to greater involvement of associative processes. To test this prediction, we let subjects watch a novel movie and later, during functional magnetic resonance imaging, asked them to rearrange and "replay" scenes from the movie in correct order. To identify areas specifically involved in retrieval of temporal order, we used a control condition where subjects logically inferred the order of scenes from the same movie. Extensive MTL activation was observed during sequence recall. Activation within the right HF was specifically related to retrieval of temporal order and correlated positively with accuracy of sequence recall. Also, the bilateral parahippocampal cortex responded to retrieval of temporal order, but the activation here was not related to performance. Our study is the first to unequivocally demonstrate that correct sequence recall depends on HF. PMID- 19295152 TI - Specificity and sufficiency of EphB1 in driving the ipsilateral retinal projection. AB - At the optic chiasm, retinal ganglion cell (RGC) axons make the decision to either avoid or traverse the midline, a maneuver that establishes the binocular pathways. In mice, the ipsilateral retinal projection arises from RGCs in the peripheral ventrotemporal (VT) crescent of the retina. These RGCs express the guidance receptor EphB1, which interacts with ephrin-B2 on radial glia cells at the optic chiasm to repulse VT axons away from the midline and into the ipsilateral optic tract. However, because VT RGCs express more than one EphB receptor, the sufficiency and specificity of the EphB1 receptor in directing the ipsilateral projection is unclear. In this study, we use in utero retinal electroporation to demonstrate that ectopic EphB1 expression can redirect RGCs with a normally crossed projection to an ipsilateral trajectory. Moreover, EphB1 is specifically required for rerouting RGC projections ipsilaterally, because introduction of the highly similar EphB2 receptor is much less efficient in redirecting RGC fibers, even when expressed at higher surface levels. Introduction of EphB1-EphB2 chimeric receptors into RGCs reveals that both extracellular and juxtamembrane domains of EphB1 are required to efficiently convert RGC projections ipsilaterally. Together, these data describe for the first time functional differences between two highly similar Eph receptors at a decision point in vivo, with EphB1 displaying unique properties that efficiently drives the uncrossed retinal projection. PMID- 19295154 TI - Evidence for a proprioception-based rapid on-line error correction mechanism for hand orientation during reaching movements in blind subjects. AB - The contribution of visual experience to the perception and sensorimotor control of spatial orientation of the hand was investigated in blind subjects. In "orientation-matching" tasks, subjects aligned a match handle held in their right hand to a target handle held in their left hand and fixed in different orientations, with both arms outstretched. In "letter-posting" task 1, the same subjects reached out and simultaneously oriented their right hand to insert the match handle into a target slot fixed in the same range of orientations. Orientations were signaled proprioceptively by a reference handle held in the left hand. Final hand orientation errors were smaller when blind subjects simultaneously reached out and rotated their hand to insert the match handle into the target slot in letter-posting task 1 than when they held their arm extended and aligned the handles in the orientation-matching task. In letter-posting task 2, blind subjects first aligned their hand to the orientation of the target and then subsequently reached to the target with the instruction to not change hand orientation during reaching. Despite the instruction, subjects showed a reduction in absolute hand orientation error from the beginning to the end of the reach. In all tasks, performance of blind subjects was very similar to that of blindfolded normally sighted subjects. These findings provide the first evidence of an automatic on-line error-correction mechanism for hand orientation guided only by proprioceptive inputs during reaching in blind subjects, and reveal that the on line mechanism does not depend on prior visual experience. PMID- 19295155 TI - Cellular plasticity for group I mGluR-mediated epileptogenesis. AB - Stimulation of group I metabotropic glutamate receptors (mGluRs) by the agonist (S)-dihydroxyphenylglycine in the hippocampus transforms normal neuronal activity into prolonged epileptiform discharges. The conversion is long lasting in that epileptiform discharges persist after washout of the inducing agonist and serves as a model of epileptogenesis. The group I mGluR model of epileptogenesis took on special significance because epilepsy associated with fragile X syndrome (FXS) may be caused by excessive group I mGluR signaling. At present, the plasticity mechanism underlying the group I mGluR-mediated epileptogenesis is unknown. I(mGluR(V)), a voltage-gated cationic current activated by group I mGluR agonists in CA3 pyramidal cells in the hippocampus, is a possible candidate. I(mGluR(V)) activation is associated with group I mGluR agonist-elicited epileptiform discharges. For I(mGluR(V)) to play a role in epileptogenesis, long-term activation of the current must occur after group I mGluR agonist exposure or synaptic stimulation. We observed that I(mGluR(V)), once induced by group I mGluR agonist stimulation in CA3 pyramidal cells, remained undiminished for hours after agonist washout. In slices prepared from FXS model mice, repeated stimulation of recurrent CA3 pyramidal cell synapses, effective in eliciting mGluR-mediated epileptiform discharges, also induced long-lasting I(mGluR(V)) in CA3 pyramidal cells. Similar to group I mGluR-mediated prolonged epileptiform discharges, persistent I(mGluR(V)) was no longer observed in preparations pretreated with inhibitors of tyrosine kinase, of extracellular signal-regulated kinase 1/2, or of mRNA protein synthesis. The results indicate that I(mGluR(V)) is an intrinsic plasticity mechanism associated with group I mGluR-mediated epileptogenesis. PMID- 19295156 TI - Diffusion-weighted magnetic resonance imaging reversal by gene knockdown of matrix metalloproteinase-9 activities in live animal brains. AB - The involvement of matrix metalloproteinase-9 (MMP-9) activities in the development of abnormal water diffusion in the brain after cardiac arrest is not fully understood. We used magnetic resonance imaging to determine the correlation between MMP-9 activity and the mechanism of abnormal water diffusion after global cerebral ischemia (GCI)-induced brain damage in C57black6 mice. We induced GCI in mice by occluding both carotid arteries for 60 min, then allowing reperfusion. We labeled a short DNA that targets mmp-9 mRNA activity [phosphorothioate-modified oligodeoxynucleotide (sODN)-mmp9] or a control probe without intracellular target (sODN-Ran) with iron-based MR contrast agent [superparamagnetic iron oxide nanoparticle (SPION)-mmp9 or SPION-Ran] or fluorescein isothiocyanate (FITC)-sODN mmp9 or FITC-sODN-Ran; we then delivered these probes by intracerebroventricular infusion or intraperitoneal injection within 3 h of reperfusion. At low dose (120 pmol/kg) the SPION-mmp9 probe was retained at significant levels in the striatum and cortex of living brains 10 h after GCI. Probe retention was validated by similar elevation of mmp-9 mRNA and antigens in postmortem samples taken from regions that exhibited GCI-induced hyperintensity in diffusion-weighted imaging, and a significant reduction in apparent diffusion coefficient (rADC, p = 0.0006, n = 12). At a higher dose (120 nmol/kg), the FITC-sODN-mmp9 probe revealed significant knockdown of MMP-9 activity, per zymography, and a reversal of striatal rADC (p = 0.004, n = 6). These observations were not duplicated in the control group. We conclude that expression of mmp-9 mRNA is associated with abnormal ADC after GCI. PMID- 19295157 TI - P2X4-receptor-mediated synthesis and release of brain-derived neurotrophic factor in microglia is dependent on calcium and p38-mitogen-activated protein kinase activation. AB - Microglia in the dorsal horn of the spinal cord are increasingly recognized as being crucial in the pathogenesis of pain hypersensitivity after injury to a peripheral nerve. It is known that P2X4 purinoceptors (P2X4Rs) cause the release of brain-derived neurotrophic factor (BDNF) from microglia, which is necessary for maintaining pain hypersensitivity after nerve injury. However, there is a critical gap in understanding how activation of microglial P2X4Rs leads to the release of BDNF. Here, we show that stimulating P2X4Rs with ATP evokes a biphasic release of BDNF from microglia: an early phase occurs within 5 min, whereas a late phase peaks 60 min after ATP stimulation. Concomitant with the late phase of release is an increased level of BDNF within the microglia. Both phases of BDNF release and the accumulation within the microglia are dependent on extracellular Ca(2+). The late phase of BDNF release and accumulation, but not the early phase of release, are suppressed by inhibiting transcription and translation, indicating that activation of P2X4R causes an initial release of a pre-existing pool of BDNF followed by an increase in de novo synthesis of BDNF. The release of BDNF is abolished by inhibiting SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)-mediated exocytosis. Furthermore, we find that the P2X4R-evoked release and synthesis of BDNF are dependent on activation of p38 mitogen-activated protein kinase (MAPK). Together, our findings provide a unifying mechanism for pain hypersensitivity after peripheral nerve injury through P2X4R-evoked increase in Ca(2+) and activation of p38-MAPK leading to the synthesis and exocytotic release of BDNF from microglia. PMID- 19295158 TI - Nuclear factor kappa B signaling regulates neuronal morphology and cocaine reward. AB - Although chronic cocaine-induced changes in dendritic spines on nucleus accumbens (NAc) neurons have been correlated with behavioral sensitization, the molecular pathways governing these structural changes, and their resulting behavioral effects, are poorly understood. The transcription factor, nuclear factor kappa B (NFkappaB), is rapidly activated by diverse stimuli and regulates expression of many genes known to maintain cell structure. Therefore, we evaluated the role of NFkappaB in regulating cocaine-induced dendritic spine changes on medium spiny neurons of the NAc and the rewarding effects of cocaine. We show that chronic cocaine induces NFkappaB-dependent transcription in the NAc of NFkappaB-Lac transgenic mice. This induction of NFkappaB activity is accompanied by increased expression of several NFkappaB genes, the promoters of which show chromatin modifications after chronic cocaine exposure consistent with their transcriptional activation. To study the functional significance of this induction, we used viral-mediated gene transfer to express either a constitutively active or dominant-negative mutant of Inhibitor of kappa B kinase (IKKca or IKKdn), which normally activates NFkappaB signaling, in the NAc. We found that activation of NFkappaB by IKKca increases the number of dendritic spines on NAc neurons, whereas inhibition of NFkappaB by IKKdn decreases basal dendritic spine number and blocks the increase in dendritic spines after chronic cocaine. Moreover, inhibition of NFkappaB blocks the rewarding effects of cocaine and the ability of previous cocaine exposure to increase an animal's preference for cocaine. Together, these studies establish a direct role for NFkappaB pathways in the NAc to regulate structural and behavioral plasticity to cocaine. PMID- 19295159 TI - Evolutionary conservation of vertebrate blood-brain barrier chemoprotective mechanisms in Drosophila. AB - Pharmacologic remedy of many brain diseases is difficult because of the powerful drug exclusion properties of the blood-brain barrier (BBB). Chemical isolation of the vertebrate brain is achieved through the highly integrated, anatomically compact and functionally overlapping chemical isolation processes of the BBB. These include functions that need to be coordinated between tight diffusion junctions and unidirectionally acting xenobiotic transporters. Understanding of many of these processes has been hampered, because they are not well mimicked by ex vivo models of the BBB and have been experimentally difficult and expensive to disentangle in intact rodent models. Here we show that the Drosophila melanogaster (Dm) humoral/CNS barrier conserves the xenobiotic exclusion properties found in the vertebrate vascular endothelium. We characterize a fly ATP binding cassette (ABC) transporter, Mdr65, that functions similarly to mammalian xenobiotic BBB transporters and show that varying its levels solely in the Dm BBB changes the inherent sensitivity of the barrier to cytotoxic pharmaceuticals. Furthermore, we demonstrate orthologous function between Mdr65 and vertebrate ABC transporters by rescuing chemical protection of the Dm brain with human MDR1/Pgp. These data indicate that the ancient origins of CNS chemoprotection extend to both conserved molecular means and functionally analogous anatomic spaces that together promote CNS selective drug partition. Thus, Dm presents an experimentally tractable system for analyzing physiological properties of the BBB in an intact organism. PMID- 19295160 TI - Disruption of Cdk5-associated phosphorylation of residue threonine-161 of the delta-opioid receptor: impaired receptor function and attenuated morphine antinociceptive tolerance. AB - Morphine is the most commonly used and most effective analgesic in the clinic. However, its use is limited by the tolerance. Evidence indicates that the delta opioid receptor (DOR) is essential for morphine antinociceptive tolerance; however, their underlying mechanisms are poorly understood. Here, we show that cyclin-dependent kinase 5 (Cdk5), activated in morphine antinociceptive tolerance, directly phosphorylates DOR at Thr-161 in DRG neurons. Cdk5 was found to phosphorylate Thr-161 in the second loop of DOR, but not the corresponding residue in the mu-opioid receptor (MOR). Phosphorylation at Thr-161 is required for normal cell surface expression of DOR, and the formation of DOR-MOR heterodimers. Our studies indicated that inhibition of Cdk5 activity or overexpression of a DOR mutant lacking the Cdk5 phosphorylation site displayed relatively low cell surface expression and relatively low abilities to form heterodimers of DOR and MOR; intrathecal delivery of a construct expressing the T161A mutant of DOR attenuated morphine antinociceptive tolerance in rats, suggesting that Thr-161 phosphorylation of DOR contributed to Cdk5-mediated morphine antinociceptive tolerance. Furthermore, an engineered Tat fusion interfering peptide corresponding to the second intracellular loop of DOR (Tat DOR-2L), reduced the cell surface expression of DOR, disrupted the formation of DOR-MOR heterodimers, and significantly attenuated the development of morphine antinociceptive tolerance after intrathecal injection. The present study indicates that Cdk5-mediated phosphorylation of DOR at Thr-161 plays a crucial role in the development of morphine tolerance and suggests the possibility of targeting DOR phosphorylation at Thr-161 to attenuate morphine antinociceptive tolerance during pain management. PMID- 19295161 TI - The cleavage products of amyloid-beta precursor protein are sorted to distinct carrier vesicles that are independently transported within neurites. AB - The amyloid-beta (Abeta) precursor protein (APP), a transmembrane protein that undergoes proteolytic cleavage into defined fragments, has been implicated in axonal transport. The proposed role of APP as a vesicle receptor for the microtubule motor kinesin-1 has relevance for the pathogenesis of Alzheimer's disease. Nevertheless, this function, which relies on the transport to the cell periphery of full-length APP rather than its cleavage fragments, remains controversial. Other proposed functions of APP, such as regulating transcription, neurogenesis, cell movement, or neurite growth also rely on APP's presence as a full-length protein at the cell surface, implying that APP cleavage occurs after its transport to the cell periphery. To test this hypothesis, we mapped the localization of various APP epitopes in neurons in culture and in the mouse brain. Surprisingly, epitopes from the N-terminal, C-terminal, and central (Abeta) domains of APP each showed a distinct distribution throughout the cell and rarely colocalized. Within neurites, these epitopes were localized to distinct transport vesicles that associated with different sets of microtubules and, occasionally, actin filaments. C-terminal APP fragments were preferentially transported into neurites as phosphorylated forms, entered the lamellipodium and filopodia of growth cones, and concentrated in regions of growth cone turning and advancement (unlike the N-terminal and Abeta fragments). We conclude that, under normal conditions, the proteolytic cleavage of APP primarily occurs before its sorting into axonal transport vesicles and the cleaved fragments segregate into separate vesicle populations that reach different destinations, and thus have different functions. PMID- 19295162 TI - Specific loss of brain ABCA1 increases brain cholesterol uptake and influences neuronal structure and function. AB - The expression of the cholesterol transporter ATP-binding cassette transporter A1 (ABCA1) in the brain and its role in the lipidation of apolipoproteins indicate that ABCA1 may play a critical role in brain cholesterol metabolism. To investigate the role of ABCA1 in brain cholesterol homeostasis and trafficking, we characterized mice that specifically lacked ABCA1 in the CNS, generated using the Cre/loxP recombination system. These mice showed reduced plasma high-density lipoprotein (HDL) cholesterol levels associated with decreased brain cholesterol content and enhanced brain uptake of esterified cholesterol from plasma HDL. Increased levels of HDL receptor SR-BI in brain capillaries and apolipoprotein A I in brain and CSF of mutant mice were evident. Cholesterol homeostasis changes were mirrored by disturbances in motor activity and sensorimotor function. Changes in synaptic ultrastructure including reduced synapse and synaptic vesicle numbers were observed. These data show that ABCA1 is a key regulator of brain cholesterol metabolism and that disturbances in cholesterol transport in the CNS are associated with structural and functional deficits in neurons. Moreover, our findings also demonstrate that specific changes in brain cholesterol metabolism can lead to alterations in cholesterol uptake from plasma to brain. PMID- 19295163 TI - Regulation of group I metabotropic glutamate receptor trafficking and signaling by the caveolar/lipid raft pathway. AB - Endocytic trafficking of neurotransmitter receptors is critical to neuronal signaling and activity-dependent synaptic plasticity. Although the importance of clathrin-mediated endocytosis in receptor trafficking in neurons is well established, the contribution of the caveolar/lipid raft pathway has been little explored. Here, we show that caveolin-1, an adaptor protein that associates with lipid rafts and the main coat protein of caveolae, binds to and colocalizes with metabotropic glutamate receptors 1/5 (mGluR1/5). The interaction with caveolin-1 controls the rate of constitutive mGluR1 internalization, thereby regulating expression of the receptor at the cell surface. Consistent with a role for caveolin-1 in mGluR trafficking, we show that mGluR1/5 associate with lipid rafts in the brain and that their constitutive internalization is mediated, in both heterologous cells and neurons, by caveolar/raft-dependent endocytosis. We further show that caveolin-1 attenuates mGluR1-dependent activation of extracellular signal-regulated kinase (ERK)-mitogen-activated protein kinase (MAPK) signaling, an effect that is abolished in cells expressing mutant mGluR1 lacking intact caveolin binding motifs. Neurons from caveolin-1 knock-out mice show enhanced basal ERK1/2 phosphorylation and prolonged ERK1/2 activation in response to stimulation with DHPG [(RS)-3,5-dihydroxyphenylglycine], a group I mGluR-selective agonist. Together, these findings underscore the importance of caveolar rafts in neurons and suggest that this pathway might play an important role in synapse formation and plasticity. PMID- 19295164 TI - Macrophage-mediated degradation of beta-amyloid via an apolipoprotein E isoform dependent mechanism. AB - Recent studies suggest that bone marrow-derived macrophages can effectively reduce beta-amyloid (Abeta) deposition in brain. To further elucidate the mechanisms by which macrophages degrade Abeta, we cultured murine macrophages on top of Abeta plaque-bearing brain sections from transgenic mice expressing PDAPP [human amyloid precursor protein (APP) with the APP(717V>F) mutation driven by the platelet-derived growth factor promoter]. Using this ex vivo assay, we found that macrophages from wild-type mice very efficiently degrade both soluble and insoluble Abeta in a time-dependent manner and markedly eliminate thioflavine-S positive amyloid deposits. Because macrophages express and secrete apolipoprotein E (apoE), we compared the efficiency of Abeta degradation by macrophages prepared from apoE-deficient mice or mice expressing human apoE2, apoE3, or apoE4. Macrophages expressing apoE2 were more efficient at degrading Abeta than apoE3 expressing, apoE4-expressing, or apoE-deficient macrophages. Moreover, macrophage induced degradation of Abeta was effectively blocked by an anti-apoE antibody and receptor-associated protein, an antagonist of the low-density lipoprotein (LDL) receptor family, suggesting involvement of LDL receptors. Measurement of matrix metalloproteinase-9 (MMP-9) activity in the media from human apoE-expressing macrophages cocultured with Abeta-containing brain sections revealed greater levels of MMP-9 activity in apoE2-expressing than in either apoE3- or apoE4 expressing macrophages. Differences in MMP-9 activity appear to contribute to the isoform-specific differences in Abeta degradation by macrophages. These apoE isoform-dependent effects of macrophages on Abeta degradation suggest a novel "peripheral" mechanism for Abeta clearance from brain that may also, in part, explain the isoform-dependent effects of apoE in determining the genetic risk for Alzheimer's disease. PMID- 19295165 TI - Adrenergic and noradrenergic innervation of the midbrain ventral tegmental area and retrorubral field: prominent inputs from medullary homeostatic centers. AB - Adrenergic agents modulate the activity of midbrain ventral tegmental area (VTA) neurons. However, the sources of noradrenergic and adrenergic inputs are not well characterized. Immunostaining for dopamine beta-hydroxylase revealed fibers within dopamine (DA) neuron areas, with the highest density in the retrorubral field (A8 cell group), followed by the VTA (A10 cell group), and very few fibers within substantia nigra compacta. A less dense, but a similar pattern of fibers was also found for the epinephrine marker, phenylethanolamine N-methyl transferase. Injection of the retrograde tracer wheat germ agglutinin-apo (inactivated) horseradish peroxidase conjugated to colloidal gold, or cholera toxin subunit b, revealed that the noradrenergic innervation of the A10 and A8 regions arise primarily from A1, A2, A5, and locus ceruleus neurons. Selective lesions of the ventral noradrenergic bundle confirmed a prominent innervation from A1 and A2 areas. Retrogradely labeled epinephrine neurons were found mainly in the C1 area. The identification of medullary noradrenergic and adrenergic afferents to DA neuron areas indicates new pathways for visceral-related inputs to reward-related areas in the midbrain. PMID- 19295166 TI - Behavioral and neural changes after gains and losses of conditioned reinforcers. AB - Human behaviors can be more powerfully influenced by conditioned reinforcers, such as money, than by primary reinforcers. Moreover, people often change their behaviors to avoid monetary losses. However, the effect of removing conditioned reinforcers on choices has not been explored in animals, and the neural mechanisms mediating the behavioral effects of gains and losses are not well understood. To investigate the behavioral and neural effects of gaining and losing a conditioned reinforcer, we trained rhesus monkeys for a matching pennies task in which the positive and negative values of its payoff matrix were realized by the delivery and removal of a conditioned reinforcer. Consistent with the findings previously obtained with non-negative payoffs and primary rewards, the animal's choice behavior during this task was nearly optimal. Nevertheless, the gain and loss of a conditioned reinforcer significantly increased and decreased, respectively, the tendency for the animal to choose the same target in subsequent trials. We also found that the neurons in the dorsomedial frontal cortex, dorsal anterior cingulate cortex, and dorsolateral prefrontal cortex often changed their activity according to whether the animal earned or lost a conditioned reinforcer in the current or previous trial. Moreover, many neurons in the dorsomedial frontal cortex also signaled the gain or loss occurring as a result of choosing a particular action as well as changes in the animal's behaviors resulting from such gains or losses. Thus, primate medial frontal cortex might mediate the behavioral effects of conditioned reinforcers and their losses. PMID- 19295167 TI - Classification of NPY-expressing neocortical interneurons. AB - Neuropeptide Y (NPY) is an abundant neuropeptide of the neocortex involved in numerous physiological and pathological processes. Because of the large electrophysiological, molecular, and morphological diversity of NPY-expressing neurons their precise identity remains unclear. To define distinct populations of NPY neurons we characterized, in acute slices of rat barrel cortex, 200 cortical neurons of layers I-IV by means of whole-cell patch-clamp recordings, biocytin labeling, and single-cell reverse transcriptase-PCR designed to probe for the expression of well established molecular markers for cortical neurons. To classify reliably cortical NPY neurons, we used and compared different unsupervised clustering algorithms based on laminar location and electrophysiological and molecular properties. These classification schemes confirmed that NPY neurons are nearly exclusively GABAergic and consistently disclosed three main types of NPY-expressing interneurons. (1) Neurogliaform-like neurons exhibiting a dense axonal arbor, were the most frequent and superficial, and substantially expressed the neuronal isoform of nitric oxide synthase. (2) Martinotti-like cells characterized by an ascending axon ramifying in layer I coexpressed somatostatin and were the most excitable type. (3) Among fast-spiking and parvalbumin-positive basket cells, NPY expression was correlated with pronounced spike latency. By clarifying the diversity of cortical NPY neurons, this study establishes a basis for future investigations aiming at elucidating their physiological roles. PMID- 19295168 TI - The cause of the imbalance in the neuronal network leading to seizure activity can be predicted by the electrographic pattern of the seizure onset. AB - This study investigates the temporal dynamics of ictal electrical activity induced by injection of the GABA(A) receptor antagonist bicuculline, and the glutamate agonist kainic acid, into the CA3 area of hippocampus. Experiments were conducted in freely moving adult Wistar rats implanted with microelectrodes in multiple brain areas. Wide-band electrical activity (0.1-3000 Hz) was recorded, and the latency of seizure onset as well as the pattern of electrical activity were investigated for each drug. The latencies between injection and the occurrence of first epileptiform events were 3.93 +/- 2.76 (+/-STD) min for bicuculline and 6.37 +/- 7.66 min for kainic acid, suggesting the existence of powerful seizure-suppressive mechanisms in the brain. Bicuculline evoked high amplitude rhythmic epileptiform events at the site of injection which resembled interictal EEG spikes and rapidly propagated to adjacent and remote brain areas. Kainic acid evoked a completely different pattern with a gradual increase in the amplitude of 30-80 Hz activity. Whereas there was strong temporal correlation between EEG events at the site of bicuculline injection and discharges in distant areas, much less correlation was seen with kainic acid injection. Both patterns were followed by generalized ictal EEG discharges and behavioral seizures. Our results illustrate that the same area of the brain can trigger seizures with different electrographic patterns. The knowledge of the network mechanisms underlying these two distinct electrographic patterns might be helpful in designing differential strategies for preventing seizure occurrence. PMID- 19295169 TI - Inhibition of Kv1.3 potassium current by phosphoinositides and stromal-derived factor-1alpha in Jurkat T cells. AB - The activation of Kv1.3 potassium channel has obligatory roles in immune responses of T lymphocytes. Stromal cell-derived factor-1alpha (SDF-1alpha) binds to C-X-C chemokine receptor type 4, activates phosphoinositide 3-kinase, and plays essential roles in cell migration of T lymphocytes. In this study, the effects of phosphoinositides and SDF-1alpha on Kv1.3 current activity were examined in the Jurkat T cell line using whole cell patch-clamp techniques. The internal application of 10 microM phosphatidylinositol 4,5-bisphosphate (PIP(2)) or 10 microM phosphatidylinositol-3,4,5-trisphosphate (PIP(3)) significantly reduced Kv1.3 current, but that of 10 microM phosphatidylinositol-4-monophosphate (PIP) did not. The coapplication of 10 microg/ml anti-PIP(3) antibody with PIP(2) from the pipette did not change the reduction of Kv1.3 current by PIP(2), but the coapplication of the antibody with PIP(3) eliminated the reduction. The heat inactivated anti-PIP(3) antibody had no effect on PIP(3)-induced inhibition. These results suggest that PIP(2) per se can reduce Kv1.3 current as well as PIP(3). External application of 1 muM Akt-kinase inhibitor VIII did not reverse the effect of intracellular PIP(3). External application of 10 and 30 ng/ml SDF 1alpha significantly reduced Kv1.3 current. Internal application of anti-PIP(3) antibody reversed the SDF-1alpha-induced reduction. These results suggest that, in Jurkat T cells, PIP(2), PIP(3), and SDF-1alpha reduce Kv1.3 channel activity and that the reduction by SDF-1alpha may be mediated by the enhancement of PIP(3) production. These novel inhibitory effects of phosphoinositides and SDF-1alpha on Kv1.3 current may have a significant function as a downregulation mechanism of Kv1.3 activity for the maintenance of T lymphocyte activation in immune responses. PMID- 19295170 TI - Loss of function of Sco1 and its interaction with cytochrome c oxidase. AB - Sco1 and Sco2 are mitochondrial copper-binding proteins involved in the biogenesis of the Cu(A) site in the cytochrome c oxidase (CcO) subunit Cox2 and in the maintenance of cellular copper homeostasis. Human Surf1 is a CcO assembly factor with an important but poorly characterized role in CcO biogenesis. Here, we analyzed the impact on CcO assembly and tissue copper levels of a G132S mutation in the juxtamembrane region of SCO1 metallochaperone associated with early onset hypertrophic cardiomyopathy, encephalopathy, hypotonia, and hepatopathy, assessed the total copper content of various SURF1 and SCO2 deficient tissues, and investigated the possible physical association between CcO and Sco1. The steady-state level of mutant Sco1 was severely decreased in the muscle mitochondria of the SCO1 patient, indicating compromised stability and thus loss of function of the protein. Unlike the wild-type variant, residual mutant Sco1 appeared to migrate exclusively in the monomeric form on blue native gels. Both the activity and content of CcO were reduced in the patient's muscle to approximately 10-20% of control values. SCO1-deficient mitochondria showed accumulation of two Cox2 subcomplexes, suggesting that Sco1 is very likely responsible for a different posttranslational aspect of Cox2 maturation than Sco2. Intriguingly, the various SURF1-deficient samples analyzed showed a tissue specific copper deficiency similar to that of SCO-deficient samples, suggesting a role for Surf1 in copper homeostasis regulation. Finally, both blue native immunoblot analysis and coimmunoprecipitation revealed that a fraction of Sco1 physically associates with the CcO complex in human muscle mitochondria, suggesting a possible direct relationship between CcO and the regulation of cellular copper homeostasis. PMID- 19295171 TI - CapZ dynamics are altered by endothelin-1 and phenylephrine via PIP2- and PKC dependent mechanisms. AB - One of the unanswered questions in muscle hypertrophy is how new contractile units are inserted into a stable existing cytoskeletal meshwork. Regulation of actin capping by CapZ may play a role in remodeling processes, therefore, CapZ dynamics are determined during rapid growth of cardiac cells in vitro. Neonatal rat ventricular myocytes were infected with adenovirus expressing green fluorescent protein-CapZ beta1 and responded normally to hypertrophic stimuli. CapZ dynamics were analyzed by fluorescence recovery after photobleaching in cultured myocytes treated with endothelin-1 (100 nM) or phenylephrine (10 muM). Recovery by 30 s was greater with endothelin treatment. Analysis 30 min postbleach showed CapZ-infected cells treated with endothelin recovered more completely than controls (77 +/- 9% vs. 50 +/- 6%, P < 0.001). Similar results were found with phenylephrine (77 +/- 5%, P < 0.05). A potential mechanism for phosphatidylinositol bisphosphate (PIP2) mediation of increased CapZ exchange in endothelin- and phenylephrine-treated cells was tested. PIP2 sequestration with neomycin (500 muM) blocked both endothelin- (43 +/- 6%, P < 0.001) and phenylephrine (36 +/- 4%, P < 0.001)-mediated recovery. The protein kinase C inhibitor chelerythrine chloride (10 muM) also blocked endothelin- (53 +/- 10%, P < 0.001) and phenylephrine (42 +/- 3%, P < 0.001)-mediated recovery. This study demonstrates for the first time that endothelin and phenylephrine alter CapZ dynamics through PIP2- and PKC-dependent pathways, which might destabilize the existing framework and permit sarcomeric remodelling to proceed. PMID- 19295172 TI - Reduced thin filament length in nebulin-knockout skeletal muscle alters isometric contractile properties. AB - Nebulin (NEB) is a large, rod-like protein believed to dictate actin thin filament length in skeletal muscle. NEB gene defects are associated with congenital nemaline myopathy. The functional role of NEB was investigated in gastrocnemius muscles from neonatal wild-type (WT) and NEB knockout (NEB-KO) mice, whose thin filaments have uniformly shorter lengths compared with WT mice. Isometric stress production in NEB-KO skeletal muscle was reduced by 27% compared with WT skeletal muscle on postnatal day 1 and by 92% on postnatal day 7, consistent with functionally severe myopathy. NEB-KO muscle was also more susceptible to a decline in stress production during a bout of 10 cyclic isometric tetani. Length-tension properties in NEB-KO muscle were altered in a manner consistent with reduced thin filament length, with length-tension curves from NEB-KO muscle demonstrating a 7.4% narrower functional range and an optimal length reduced by 0.13 muscle lengths. Expression patterns of myosin heavy chain isoforms and total myosin content did not account for the functional differences between WT and NEB-KO muscle. These data indicate that NEB is essential for active stress production, maintenance of functional integrity during cyclic activation, and length-tension properties consistent with a role in specifying normal thin filament length. Continued analysis of NEB's functional properties will strengthen the understanding of force transmission and thin filament length regulation in skeletal muscle and may provide insights into the molecular processes that give rise to nemaline myopathy. PMID- 19295173 TI - Mitogen-activated protein kinase signaling is necessary for the maintenance of skeletal muscle mass. AB - The signal transduction cascades that maintain muscle mass remain to be fully defined. Herein, we report that inhibition of extracellular signal-regulated kinase 1/2 (ERK1/2) signaling in vitro decreases myotube size and protein content after 3-day treatment with a MEK inhibitor. Neither p38 nor JNK inhibitors had any effect on myotube size or morphology. ERK1/2 inhibition also upregulated gene transcription of atrogin-1 and muscle-specific RING finger protein 1 and downregulated the phosphorylation of Akt and its downstream kinases. Forced expression of enhanced green fluorescent protein-tagged MAPK phosphatase 1 (MKP 1) in soleus and gastrocnemius muscles decreased both fiber size and reporter activity. This atrophic effect of MKP-1 was time dependent. Analysis of the reporter activity in vivo revealed that the activities of nuclear factor-kappaB and 26S proteasome were differentially activated in slow and fast muscles, suggesting muscle type-specific mechanisms may be utilized. Together, these findings suggest that MAPK signaling is necessary for the maintenance of skeletal muscle mass because inhibition of these signaling cascades elicits muscle atrophy in vitro and in vivo. PMID- 19295174 TI - Endogenous expression of TRPV5 and TRPV6 calcium channels in human leukemia K562 cells. AB - In blood cells, changes in intracellular Ca(2+) concentration ([Ca(2+)](i)) are associated with multiple cellular events, including activation of cellular kinases and phosphatases, degranulation, regulation of cytoskeleton binding proteins, transcriptional control, and modulation of surface receptors. Although there is no doubt as to the significance of Ca(2+) signaling in blood cells, there is sparse knowledge about the molecular identities of the plasmalemmal Ca(2+) permeable channels that control Ca(2+) fluxes across the plasma membrane and mediate changes in [Ca(2+)](i) in blood cells. Using RNA expression analysis, we have shown that human leukemia K562 cells endogenously coexpress transient receptor potential vanilloid channels type 5 (TRPV5) and type 6 (TRPV6) mRNAs. Moreover, we demonstrated that TRPV5 and TRPV6 channel proteins are present in both the total lysates and the crude membrane preparations from leukemia cells. Immunoprecipitation revealed that a physical interaction between TRPV5 and TRPV6 may take place. Single-channel patch-clamp experiments demonstrated the presence of inwardly rectifying monovalent currents that displayed kinetic characteristics of unitary TRPV5 and/or TRPV6 currents and were blocked by extracellular Ca(2+) and ruthenium red. Taken together, our data strongly indicate that human myeloid leukemia cells coexpress functional TRPV5 and TRPV6 calcium channels that may interact with each other and contribute into intracellular Ca(2+) signaling. PMID- 19295175 TI - Amantadine inhibits platelet-activating factor induced clathrin-mediated endocytosis in human neutrophils. AB - Receptor signaling is integral for adhesion, emigration, phagocytosis, and reactive oxygen species production in polymorphonuclear neutrophils (PMNs). Priming is an important part of PMN emigration, but it can also lead to PMN mediated organ injury in the host. Platelet-activating factor (PAF) primes PMNs through activation of a specific G protein-coupled receptor. We hypothesize that PAF priming of PMNs requires clathrin-mediated endocytosis (CME) of the PAF receptor (PAFr), and, therefore, amantadine, known to inhibit CME, significantly antagonizes PAF signaling. PMNs were isolated by standard techniques to >98% purity and tested for viability. Amantadine (1 mM) significantly inhibited the PAF-mediated changes in the cellular distribution of clathrin and the physical colocalization [fluorescence resonance energy transfer positive (FRET+)] of early endosome antigen-1 and Rab5a, known components of CME and similar to hypertonic saline, a known inhibitor of CME. Furthermore, amantadine had no effect on the PAF-induced cytosolic calcium flux; however, phosphorylation of p38 MAPK was significantly decreased. Amantadine inhibited PAF-mediated changes in PMN physiology, including priming of the NADPH oxidase and shape change with lesser inhibition of increases in CD11b surface expression and elastase release. Furthermore, rimantadine, an amantadine analog, was a more potent inhibitor of PAF priming of the N-formyl-methionyl-leucyl-phenylalanine-activated oxidase. PAF priming of PMNs requires clathrin-mediated endocytosis that is inhibited when PMNs are pretreated with either amantadine or rimantadine. Thus, amantadine and rimantadine have the potential to ameliorate PMN-mediated tissue damage in humans. PMID- 19295176 TI - Control of mitochondrial biogenesis, ROS level, and cytosolic Ca2+ concentration during the cell cycle and the onset of differentiation in L6E9 myoblasts. AB - Mitochondria can sense signals linked to changes in energy demand to affect nuclear gene expression. This retrograde signaling pathway is presumed to be involved in the regulation of myoblast proliferation and differentiation. We have investigated the regulation of mitochondrial biogenesis and production of putative retrograde signaling agents [hydrogen peroxide (H(2)O(2)) and Ca(2+)] during the cell cycle and the onset of differentiation in L6E9 muscle cells. The biosynthesis of cardiolipin and mitochondrial proteins was mainly achieved in S phase, whereas the expression of mitochondrial biogenesis factors [peroxisome proliferator-activated receptor (PPAR)-alpha, PPAR-delta, and neuronal nitric oxide synthase 1] was regularly increased from G(1) to G(2)M phase. In agreement with the increase in mitochondrial membrane potential, mitochondria in S and G(2)M phases have a significantly higher H(2)O(2) level when compared with G(1) phase. By contrast, the onset of differentiation was characterized by a marked reduction in mitochondrial protein expression and mitochondrial H(2)O(2) level. The capacity of mitochondria to release Ca(2+) in response to a metabolic challenge was significantly decreased at the onset of differentiation. Finally, an increase in calmodulin expression in S and G(2)M phases and a transitory increase in phosphorylated nuclear factor of activated T cells (NFAT) c3 in S phase was observed. NFATc3 phosphorylation was markedly decreased at the onset of differentiation. Our data point to functional links between the control of mitochondrial biogenesis and the regulation of the level of retrograde signaling agents during the cell cycle and the onset of differentiation in L6E9 muscle cells. PMID- 19295177 TI - Nongenomic actions of L-thyroxine and 3,5,3'-triiodo-L-thyronine. Focus on "L Thyroxine vs. 3,5,3'-triiodo-L-thyronine and cell proliferation: activation of mitogen-activated protein kinase and phosphatidylinositol 3-kinase". PMID- 19295178 TI - Involvement of calpains in Ca2+-induced disruption of excitation-contraction coupling in mammalian skeletal muscle fibers. AB - In skeletal muscle fibers, the coupling between excitation of the surface membrane and the release of Ca(2+) from the sarcoplasmic reticulum is irreversibly disrupted if cytoplasmic Ca(2+) concentration ([Ca(2+)]) is raised to micromolar levels for a prolonged period. This excitation-contraction (EC) uncoupling may contribute to muscle weakness after some types of exercise and in certain muscle diseases and has been linked to structural alteration of the triad junctions, but its molecular basis is unclear. Both mu-calpain, a ubiquitous Ca(2+)-activated protease, and muscle-specific calpain-3 become autolytically activated at micromolar Ca(2+) and have been suggested to be responsible for the uncoupling. This study used controlled Ca(2+) exposure in mechanically skinned fibers from extensor digitorum longus muscle to show that EC uncoupling still occurs in muscle fibers of calpain-3-deficient mice, with a Ca(2+) dependence indistinguishable from that in normal mice and rats. Western blotting of muscle fibers that had been partially EC uncoupled by exposure to an intermediate Ca(2+) level ( approximately 5 microM Ca(2+) for 3 min, no ATP) showed the presence of autolytic activation of a proportion of the mu-calpain present, but with little or no activation of calpain-3. Homogenates of normal and calpain-3-deficient muscles exposed to micromolar Ca(2+) displayed similar levels of diffusible proteolytic activity, as gauged by the rate of decline of passive force in stretched, skinned muscle fibers. Exogenously added mu-calpain, preactivated by elevated [Ca(2+)] and applied in the presence of 1 microM Ca(2+), disrupted EC coupling in a manner similar to raised [Ca(2+)]. We conclude that calpain-3 is not responsible for Ca(2+)-induced disruption of EC coupling, but that mu-calpain is a plausible candidate. PMID- 19295179 TI - Clinical and genetic studies in a Chinese family with giant axonal neuropathy. AB - The objective of the study was to investigate a girl with giant axonal neuropathy and detect the mutation of GAN gene in her family. The encoding exons of GAN gene were amplified from genomic DNA of the proband and her parents by polymerase chain reaction and directly sequenced after purification. The proband manifested typical neurological symptoms and pathological abnormalities. The case had 2 heterozygous missense mutations in GAN gene: 1. c. 224 T>A in exon 2, her mother was a heterozygote of this mutation and had normal phenotype; 2. c.1634G>A in exon 10, and her father was a heterozygote of this mutation and had normal phenotype. Both of the mutations caused amino acid changes in the gigaxonin protein. In this family, missense mutation of c.224 T>A and missense mutation of c.1634G>A in GAN gene caused the phenotype of giant axonal neuropathy in the proband. Her parents are heterozygotes of the disease without symptoms. PMID- 19295180 TI - Practice parameters in child neurology: do pediatricians use them? AB - We assessed pediatrician awareness of the parameter ''Evaluating the first non febrile seizure in children'' and how the concepts of this parameter were incorporated into practice. Although most reported caring for children with seizures, 60% were not aware of the practice parameter. When given the clinical scenario of an otherwise healthy 8-year-old child with a first, unprovoked seizure, management was variable. Most (83%) would obtain an electroencephalography, and many (58%) would order an imaging study, usually a magnetic resonance imaging. However, most were also likely to order laboratory studies that were not indicated given the scenario and the practice parameter. This pilot study suggests that pediatricians may not be aware of this practice parameter and many may not be incorporating evidence-based recommendations regarding the evaluation of children with new-onset seizures. PMID- 19295181 TI - Childhood mefloquine-induced mania and psychosis: a case report. AB - Mefloquine, a commonly used oral antimalarial is occasionally associated with severe, neuropsychiatric adverse effects, especially in adults. Such events are extremely rare in children. The authors report on an 11-year-old, otherwise healthy girl from Eastern India, a malaria-endemic region, who developed mania and psychosis following intake of a therapeutic dose of mefloquine for Plasmodium falciparum malaria. She recovered satisfactorily with risperidone therapy. To our knowledge, there is only one documented instance of mefloquine-induced psychosis in the pediatric literature to date. Those caring for children need to realize that severe neuropsychiatric manifestations may be seen in the pediatric age group. A positive history of intake of the offending drug with careful exclusion of other etiologies usually clinches the diagnosis. PMID- 19295182 TI - Pharyngeal-cervical-brachial variant of pediatric Guillain-Barre syndrome with antecedent acute hepatitis A virus infection. AB - Pharyngeal-cervical-brachial weakness is considered a variant of Guillain-Barre syndrome with limited oropharyngeal, neck, and upper limb muscle involvement. The authors report on a 7-year-old boy, who developed pharyngeal-cervical-brachial type of Guillain-Barre syndrome following an antecedent episode of acute hepatitis A virus infection, 2 weeks prior to admission. The presentation was characterized by acute onset dysphagia, loss of head control, and bilateral arm weakness. The diagnosis was confirmed by acute motor axonal changes in the arm and albuminocytologic dissociation of the cerebrospinal fluid. The child was treated with intravenous immunoglobulin, which resulted in gradual improvement over 3 weeks. Documented instances of this form of Guillain-Barre syndrome remain rare in the pediatric age group, with none existing following antecedent hepatitis A virus infection. The authors emphasize that acute hepatitis A virus infection be included in the triggers responsible for Guillain-Barre syndrome in children. PMID- 19295183 TI - Dietary sucrose is essential to the development of liver injury in the methionine choline-deficient model of steatohepatitis. AB - Methionine-choline-deficient (MCD) diets cause steatohepatitis in rodents and are used to study the pathophysiology of fatty liver disease in human beings. The most widely used commercial MCD formulas not only lack methionine and choline but also contain excess sucrose and fat. The objective of this study was to determine whether dietary sucrose in the MCD formula plays a role in the pathogenesis of MCD-related liver disease. We prepared two custom MCD formulas, one containing sucrose as the principal carbohydrate and the other substituting sucrose with starch. Mice fed the sucrose-enriched formula developed typical features of MCD related liver disease, including hepatic steatosis, hepatocellular apoptosis, alanine aminotransferase elevation, lipid peroxidation, and hepatic inflammation. In contrast, mice fed MCD-starch were significantly protected against liver injury. MCD-sucrose and MCD-starch mice displayed identical diet-related abnormalities in hepatic fatty acid uptake and triglyceride secretion. Hepatic de novo lipogenesis and triglyceride synthesis, however, were 2 times higher in MCD sucrose mice than MCD-starch mice (P < 0.01). Hepatic lipid analysis revealed accumulation of excess saturated fatty acids in MCD-sucrose mice that correlated with hepatocellular injury. Overall, the results indicate that dietary sucrose is critical to the pathogenesis of MCD-mediated steatohepatitis. They suggest that saturated fatty acids, which are products of de novo lipogenesis, are mediators of hepatic toxicity in this model of liver disease. PMID- 19295185 TI - The psoriasis family index: preliminary results of validation of a quality of life instrument for family members of patients with psoriasis. AB - BACKGROUND: Psoriasis can have a major impact on the lives of patients, the members of their families and their partners. OBJECTIVES: To develop and validate a disease-specific instrument to measure the secondary impact of psoriasis on the health-related quality of life (HRQoL) of family members of psoriasis patients. METHODS: The development of the new questionnaire involved a number of different stages: qualitative interviews with family members/partners, formulation of the first draft, refinement, initial validation, further refinement and finally revalidation of the questionnaire. RESULTS: Items for the measure were generated from the content analysis of interview transcripts. The psychometric evaluation of the final draft of the Psoriasis Family Index (PFI-15) was carried out in a new cohort of participants. The PFI-15 demonstrated high internal consistency (Cronbach's alpha = 0.86) and high test-retest reliability (intraclass correlation coefficient = 0.93). The construct validity was assessed by testing a number of a priori hypotheses about its construct; a moderate to strong correlation was seen between families' PFI scores and patients' Dermatology Life Quality Index scores (r = 0.54, p < 0.01) and Psoriasis Disability Index scores (r = 0.59, p < 0.01), and between families' PFI scores and patients' Psoriasis Area and Severity Index scores (r = 0.43, p < 0.01). CONCLUSIONS: The PFI is a simple and practical measure to assess the HRQoL of family members/partners of patients with psoriasis. PMID- 19295184 TI - Suppression of superoxide anion and elastase release by C18 unsaturated fatty acids in human neutrophils. AB - The structure-activity relationship of 18-carbon fatty acids (C(18) FAs) on human neutrophil functions and their underlying mechanism were investigated. C(18) unsaturated (U)FAs potently inhibited superoxide anion production, elastase release, and Ca(2+) mobilization at concentrations of <10 microM in formyl-L methionyl-L-leucyl-L-phenylalanine (FMLP)-activated human neutrophils. However, neither saturated FA nor esterified UFAs inhibited these neutrophil functions. The inhibitory potencies of C(18) UFAs decreased in the following order: C(18):1 > C(18):2 > C(18):3 > C(18):4. Notably, the potency of attenuating Ca(2+) mobilization was closely correlated with decreasing cellular responses. The inhibitions of Ca(2+) mobilization by C(18) UFAs were not altered in a Ca(2+) containing Na(+)-deprived medium. Significantly, C(18) UFAs increased the activities of plasma membrane Ca(2+)-ATPase (PMCA) in neutrophils and isolated cell membranes. In contrast, C(18) UFAs failed to alter either the cAMP level or phosphodiesterase activity. Moreover, C(18) UFAs did not reduce extracellular Ba(2+) entry in FMLP- and thapsigargin-activated neutrophils. In summary, the inhibition of neutrophil functions by C(18) UFAs is attributed to the blockade of Ca(2+) mobilization through modulation of PMCA. We also suggest that both the free carboxy group and the number of double bonds of the C(18) UFA structure are critical to providing the potent anti-inflammatory properties in human neutrophils. PMID- 19295186 TI - KRJ-I and BON cell lines: defining an appropriate enterochromaffin cell neuroendocrine tumor model. AB - BACKGROUND: Neuroendocrine tumors (NETs) of the gastrointestinal (GI) system are increasing in incidence with minimal improvement in prognosis. Although the cell of origin has been identified as the enterochromaffin (EC) cell, its secretory and proliferative regulation has not been defined at a mechanistic level. To date, the BON cell line has been the most widely used in vitro EC cell model despite its pancreatic origin. Using whole-genome mathematical analysis as well as secretory and proliferative studies, we compared the BON cell line to the small intestine (SI) EC cell-derived NET cell line, KRJ-I, to assess individual cell line validity and applicability for the investigation of GI-NET disease. METHODS AND RESULTS: Principal component analysis and ANOVA of KRJ-I and BON transcriptomes (U133 Plus 2) identified substantially different (<10%) overlap in transcripts with minimal (R(2) = 0.24) correlation in gene expression profiles. RT-PCR detected large variability (>12%) in neuroendocrine (NE) marker transcripts in the BON cell line and the absence of Tph-2, DDC, TGFbetaR2, and M3 transcripts in KRJ-I. The KRJ-I cell line secreted serotonin (5-HT) in response to isoproterenol (EC(50) = 100 nM), noradrenaline (EC(50) = 1.7 nM), and pituitary adenylate cyclase (PACAP, EC(50) = 0.03 nM). Cholecystokinin (IC(50) = 430 nM), somatostatin (IC(50) = 400 nM), acetylcholine (IC(50) = 3.7 nM), and gamma-aminobutyric acid A (GABA(A), IC(50) = 2 nM) all inhibited 5-HT release, while gastrin and bombesin had no effect. 5-HT secretion in the BON cell line was stimulated by isoproterenol (EC(50) = 900 nM), noradrenaline (EC(50) = 20 nM), cholecystokinin (EC(50) = 130 nM), PACAP (EC(50) = 0.12 nM), bombesin (EC(50) = 15 nM), and acetylcholine (EC(50) = 0.2 nM). It was inhibited by somatostatin (IC(50) = 300 nM) but not GABA(A). KRJ-I responded with proliferation to connective tissue growth factor (CTGF, EC(50) = 0.002 ng/ml), transforming growth factor-alpha (TGFalpha, EC(50) = 0.63 ng/ml) and transforming growth factor-beta (TGFbeta, EC(50) = 0.63 ng/ml). Epidermal growth factor (EGF) and somatostatin had no significant effect. BON cell proliferation was stimulated only by EGF and TGFalpha (EC(50) = 15.8 and 10 ng/ml). TGFbeta (IC(50) = 0.16 ng/ml), MZ-4-147 (IC(50) = 0.5 nM), and BIM23A761 (IC(50) = 0.06 nM) all inhibited proliferation. CTGF and somatostatin had no effect. CONCLUSION: KRJ-I and BON cell lines demonstrate substantial differences in gene level transcripts, inconsistent receptor profile expression, wide variability in NE marker transcript levels, and significantly differential proliferative and secretory responses. Given the EC cell origin of KRJ-I, these results provide evidence that the BON cell line does not represent an EC cell system and is not a valid study model of (carcinoid) EC cell-derived NET. PMID- 19295187 TI - Diffuse bleeding from the bronchial mucosa on flexible bronchoscopy in lung transplanted patients: 'the hemorrhagic bronchial mucosa syndrome'. AB - BACKGROUND: Flexible bronchoscopy (FFB) with bronchoalveolar lavage and transbronchial lung biopsy is a frequently performed procedure after lung transplantation (LTx). During 2002-2006, we performed 1,800 FFBs on LTx patients at our center. OBJECTIVES: To describe the occurrence of bronchial mucosal bleeding during FFB in 5 patients. METHODS: FFB was performed under sedation with propofol and after intubation with a laryngeal mask. All 5 patients had normal chest X-rays and no clinical signs of lower respiratory tract infection. They were normotensive, had normal lung function, normal platelet counts and prothrombin times and no clinical evidence of other coagulopathies. Immunosuppression consisted of cyclosporin A, azathioprine and prednisolone. RESULTS: In these 5 patients, we observed diffuse bleeding from the bronchial mucosa in both the transplanted and native lung. Bleeding was preceded by coughing and the development of diffuse mucosal hyperemia followed by mucosal petechiae, which subsequently coalesced into mucosal hemorrhage and overt intrabronchial bleeding. In 1 case, the bleeding caused impairment of lung function and hypoxemia. CONCLUSION: Bronchial mucosal bleeding is probably caused by increased intrathoracic pressure induced by coughing in a closed airway system, which in turn induces a high pressure in the intrathoracic vessels that is transferred to the bronchial vascular system. Vulnerability of the mucosal capillaries due to immunosuppression probably also contributes to what we have termed 'the hemorrhagic bronchial mucosa syndrome'. The hemorrhagic bronchial mucosa syndrome with intrabronchial bleeding is a rare complication of bronchoscopy, predominantly occurring in LTx patients. From a prophylactic point of view, it seems essential to avoid coughing during bronchoscopy in intubated patients. PMID- 19295188 TI - Sex differences in brain and plasma beta-endorphin content following testosterone, dihydrotestosterone and estradiol administration to gonadectomized rats. AB - AIMS: The present study aims at evaluating the effect of a 2-week treatment with testosterone (T), dihydrotestosterone (DHT) and estradiol valerate (E(2)V) on brain and plasma beta-endorphin (beta-END) concentrations in gonadectomized rats of both sexes. METHODS: Eight groups of female and 8 groups of male Wistar rats were included. For each sex, 1 group of gonad-intact and 1 group of gonadectomized rats were employed as controls (placebo). The other groups received subcutaneous T at the doses of 10 and 100 microg/kg/day (female rats) or 1 and 5 mg/kg/day (male rats). Subcutaneous DHT was administered at the doses of 1, 10, 100 microg/kg/day (female rats) or 0.1, 1 and 5 mg/kg/day (male rats). E(2)V was administered subcutaneously at 0.05 mg/kg/day. beta-END levels were measured in different brain areas and plasma. RESULTS: Ovariectomy (OVX) induced a significant decrease in beta-END in all brain areas analyzed as well as in plasma. Orchidectomy (OCX) reduced opioid concentration in the hypothalamus, anterior pituitary and neurointermediate lobe. In OVX rats, T replacement as well as E(2)V significantly increased beta-END concentration in all brain areas and in plasma. In the OCX group, T and E(2)V did not influence beta-END concentrations in different hypothalamic areas. However, they produced a significant rise in beta-END levels in the hypothalamus, neurointermediate lobe, anterior pituitary and plasma. Conversely, DHT replacement did not affect beta-END levels at any dose administered, either in males or females. CONCLUSIONS: The sensitivity of the endogenous opiatergic system to T administration seems to be sex-related. This effect is particularly evident in the brains of female animals where this endogenous endorphin elicits a much greater response than it does in males that have undergone gonadal steroid depletion and subsequent T replacement. PMID- 19295189 TI - Treatment of mediastinal fibrosis with mycophenolate mofetil. AB - We present the case of a 67-year-old male patient with mediastinal and retroperitoneal fibrosis. In Europe, this is a rare disease. Treatment was established to prevent complications due to strictures or compressions. Because of his diabetes, a therapy of low-dose prednisone combined with mycophenolate mofetil, known as steroid sparing agent, was applied. As a result, the radiological follow-up showed a marked decrease in the mediastinal and retroperitoneal masses. PMID- 19295190 TI - Lymphoproliferative disorders after immunosuppressive therapy for aplastic anemia: a case report and literature review. AB - A 61-year-old Japanese man was referred to our hospital in 2002 due to severe pancytopenia. Bone marrow and peripheral blood findings indicated he had severe aplastic anemia (AA). A whole-body CT scan and Ga scintigraphy revealed no abnormal findings. Antithymocyte globulin and cyclosporine A (CyA) were administered and he got transfusion independently. In September 2004, he complained of abdominal fullness and a skin eruption in the lower abdomen. An abdominal CT revealed a spleen mass and lymphoadenopathy of the pancreas head. Splenectomy was done, and he was diagnosed with a diffuse large B cell lymphoma (DLBCL) of the spleen and skin. His karyotype was associated with t(14; 18). CyA was stopped, all lesions disappeared, and then his AA relapsed. In January 2007, antithymocyte globulin/CyA was readministered. In May 2007, he complained of acute swelling in his right thigh. A biopsy from the tumor revealed DLBCL. CyA was stopped again, yet the lymphoma did not regress. He was given R-CHOP (rituximab, cyclophosphamide, doxorubicin hydrochloride, vincristine, prednisolone), followed by 5 cycles of R-VP (rituximab, vincristine, prednisolone) and radiation therapy, resulting in a partial remission. We report DLBCL after immunosuppressive therapy for AA. Although this is a rare complication, it should be considered before beginning immunosuppressive therapy. PMID- 19295191 TI - Dietary intake of individual glucosinolates in participants of the EPIC Heidelberg cohort study. AB - BACKGROUND/AIMS: To compile a database on content of individual glucosinolates in food and to describe the dietary intake of individual glucosinolates in a German population. METHODS: Studies analysing the content of individual glucosinolates in food were aggregated to form a database of 26 individual glucosinolates in 18 vegetables and condiments consumed in Germany. This database was linked to food intake data derived from 24-hour diet recalls of 2,121 participants of the EPIC Heidelberg cohort study. RESULTS: Mean total glucosinolate intake (+/-standard error) was 14.2 (+/-1.1) mg/day for men and 14.8 (+/-1.3) mg/day for women. The intake increased with age and education; smokers ingested less glucosinolates than never or former smokers. The quantitatively most important individual glucosinolates were glucobrassicin and sinigrin with mean daily intakes of 3.5 (+/-0.3) and 1.7 (+/-0.2) mg/day for men, and 4.2 (+/-0.4) and 2.5 (+/-0.4) mg/day for women, respectively. Broccoli, Brussels sprouts and cauliflower contributed most to the total glucosinolate intake in this population. CONCLUSIONS: The established database allowed for the first time the estimation of dietary intake of individual glucosinolates. The database can be used for epidemiological research on the role of glucosinolates in health and disease. PMID- 19295192 TI - The impact of nutritional fatty acids during pregnancy and lactation on early human adipose tissue development. Rationale and design of the INFAT study. AB - Recent observational studies suggest that mean birth weight and body fat growth in the first year of life have increased continuously over the last decades. Both elevated birth weight and early fat mass are potential risk factors for childhood obesity. Experimental and limited clinical data suggest that the dietary ratio of n-6 to n-3 fatty acids (FAs) during pregnancy is critical for early adipose tissue growth. The aim of this randomized controlled study is to examine the effect of the supplementation with n-3 long-chain polyunsaturated FAs and reduction in the n-6/n-3 ratio in the diet of pregnant women/breast-feeding mothers on adipose tissue growth in their newborns using various methods for the assessment of body fat mass. Measurement of skinfold thickness in the newborn is the primary outcome parameter. Two hundred and four pregnant women will be recruited before the 15th week of gestation and randomly assigned to either active intervention or an isocaloric control diet. This upcoming study will explore the potential of this dietary approach to limit early adipose tissue growth and may contribute to the development of a new strategy for the primary prevention of childhood obesity. PMID- 19295193 TI - Influence of Trp64Arg polymorphism of beta 3-adrenoreceptor gene on insulin resistance, adipocytokines and weight loss secondary to two hypocaloric diets. AB - OBJECTIVE: To investigate the influence of the Trp64Arg polymorphism in the beta 3-adrenoreceptor gene on adipocytokines, insulin resistance and weight loss secondary to a low fat versus a low carbohydrate diet. METHODS: A population of 193 obese patients was analyzed. A nutritional evaluation was performed at the beginning and at the end of a 2-month period in which subjects received 1 of 2 hypocaloric diets. RESULTS: 172 (89.1%; 44 males, 128 females) subjects had the genotype Trp64/Trp64 (wild-type group) and 21 subjects (10.9%; 4 males, 17 females) had Trp64/Arg64 (mutant-type group). In the wild-type group BMI, weight, fat mass, waist circumference, systolic blood pressure, glucose, triglycerides, insulin, homeostasis model assessment (HOMA) and leptin decreased significantly regardless of the diet. In the mutant-type group, this was the case for BMI, weight, waist circumference, fat mass and leptin levels. Only leptin levels have a significant decrease in the wild-type group (diet I 13.7%, diet II 26.3%, p < 0.05 for both). In the mutant group, leptin decreased as well (diet I 22.5%, diet II 30.1%, p < 0.05 for both). CONCLUSION: The metabolic effect of mild weight reduction by 2 hypocaloric diets is greatest in subjects homozygous for the normal beta 3-adrenoreceptor gene. Improvement in glucose, insulin and HOMA is better than in the mutant-type group. PMID- 19295194 TI - Patterns and predictors of long-term weight change in patients with type 2 diabetes mellitus. AB - This study describes patterns of long-term weight changes among patients with type 2 diabetes mellitus (DM) and identifies factors associated with weight changes. During the mean follow-up period of 9.1 (SD 3.6; range 1-15) years, 7,820 patients with type 2 DM have been examined to determine weight changes. Their weight at the last clinic visit was compared with that at the initial visit. The mean age of the participants was 51.3 (SD 10.5) years, with a mean DM duration of 6.1 (SD 5.6) years at initial registration. Mean weight change was 1.9% (95% confidence interval, CI -2.1 to -1.7) and varied according to the severity of baseline obesity (-4.1%; 95% CI -4.6 to -3.6) for normal weight, 8.9% (95% CI -13.0 to -4.8) for underweight, -1.7% (95% CI -2.1 to -1.3) for overweight and -0.3% (95% CI -0.03 to +0.73) for obese, and +0.2% (95% CI -1.8 to +2.1) for morbidly obese patients. Using a stepwise multiple regression model, higher body mass index, follow-up, fasting plasma glucose, systolic blood pressure, triglyceride level and treatment with insulin increased the percent weight change, and higher number of follow-up, cholesterol and smoking significantly decreased it. Although this Iranian patients with type 2 DM had negligible weight change over 9.1 years on average, several clinical and lifestyle characteristics were associated with weight change. PMID- 19295195 TI - Undernutrition and oxidative stress as risk factors for high blood pressure in older Mexican adults. AB - OBJECTIVE: To determine the relationship between undernutrition and oxidative stress as risk factors for high blood pressure (HBP) in older Mexican adults. We carried out a case-control study in a convenience sample of 62 older adults, 60 80 years of age; 32 without HBP (mean age 75.8 +/- 5.1 years) and 30 with HBP (mean age 65.2 +/- 6.8 years). METHODS: We measured serum lipoperoxide (LPO) levels, antioxidant enzymes, total antioxidant activity and lipid profile; in addition, we evaluated the nutritional status with anthropometric measurements, the Mini Nutritional Assessment and food intake frequency. Data were analyzed with Student t and odds ratio (OR) statistical tests. RESULTS: We observed significantly higher LPO levels in subjects with HBP than in normotensive subjects (0.303 +/- 0.10 vs. 0.256 +/- 0.06 micromol/l; p < 0.05). With respect to risk factors for HBP, we found an OR of 12.2 (95% CI 1.9-76.4, p < 0.01) in subjects with high LPO levels (> or = 0.320 micromol/l) and an OR of 5.0 (95% CI 1.1-23.0, p < 0.05) in those with undernutrition. CONCLUSION: Our findings suggest that high LPO levels and undernutrition are significant risk factors for HBP. PMID- 19295196 TI - Pathophysiology of calcium and phosphate metabolism impairment in chronic kidney disease. AB - Secondary hyperparathyroidism (SHPT) is a classical feature of chronic kidney disease (CKD). Commonly, hypocalcemia, hyperphosphatemia, and vitamin D deficiency are involved into the pathogenesis of SHPT. Parathyroid (PT) glands are characterized by a low turnover and rarely undergo mitoses. However, in the presence of low calcium, high phosphorus, vitamin D deficiency, and uremia, PT cells leave quiescence. In the last decade, both new molecular and cellular mechanisms have been investigated in the pathophysiology of SHPT, between them the emerging role of the PT vitamin D receptor and calcium-sensing receptor. Furthermore, recent studies indicate that the fibroblast growth factor-23 may play a central role in the regulation of phosphate-vitamin D metabolism in CKD. Certainly, in the next future, these new insights into the pathogenesis of SHPT will give the possibility to improve the treatment of this condition in the CKD population. PMID- 19295197 TI - Dietary and pharmacological control of calcium and phosphate metabolism in predialysis stages of chronic kidney disease. AB - Data on calcium and phosphate metabolism in the predialysis stages of chronic kidney disease (CKD) are scarce when compared with the available information on patients on dialysis. Visible derangements of calcium and phosphate levels start to become apparent when GFR falls below 40 ml/min. In some but not all patients, hyperphosphatemia may be a mortality risk predictor in CKD stages 4-5. There are only few treatment studies targeting hyperphosphatemia in these CKD stages. However, the RIND study, evaluating progression of coronary artery calcification in incident hemodialysis patients, demonstrated that vascular calcification processes manifest in predialysis stages in the majority of patients, which may well be linked to deranged calcium and phosphate homeostasis. Novel insights into the pathophysiology of calcium and phosphate handling, especially the discovery of the phosphatonin FGF23, suggest that a more complex assessment of phosphate balance is warranted. This assessment should include measurements of fractional phosphate excretion and phosphatonin levels to objectively judge and effectively correct phosphate overload. PMID- 19295198 TI - Impact of disturbances of calcium and phosphate metabolism on vascular calcification and clinical outcomes in patients with chronic kidney disease. AB - Chronic kidney disease (CKD) is frequently complicated by arterial calcification. The latter is part of the associated mineral and bone disorder (CKD-MBD). Hypercalcemia and hyperphosphatemia have long been known to play a major role in the occurrence of vascular and other soft tissue calcification in patients with CKD, together with endocrine disturbances including vitamin D, parathyroid hormone, fibroblast growth factor-23, and klotho. In addition, many other systemic and local promoters, including inflammation and uremic toxins, contribute to the occurrence of vascular calcification, despite a powerful defense system made up of systemic and local inhibitors, as demonstrated in elegant experimental studies done in vitro and in vivo. Most importantly, several reports have shown that both hyperphosphatemia and hypophosphatemia, and to a lesser degree hypercalcemia and hypocalcemia, are associated with an increased relative risk of mortality in patients with CKD. However, all these reports were observational in nature and must therefore be considered as hypothesis generating. It remains to be demonstrated in prospective randomized trials whether normalization of serum phosphorus and/or calcium leads to better patient outcome. In order to improve outcome in patients with CKD-MBD, early medical intervention is of utmost importance. PMID- 19295199 TI - Calcium and phosphate control by dialysis treatments. AB - Calcium and phosphate changes, besides their involvement in bone disease, have been claimed to also be involved in the increased vascular morbidity and mortality of dialysis patients. Even after the recent advances of therapeutic options, their control still remains a challenging problem. Dialysis treatment is a basic approach to the control of these two electrolytes. Calcium control by dialysis is mainly dependent on its mass balance, which is variably influenced by the calcium concentration difference between blood and dialysis solutions (either dialysate or infusion fluids) and by the duration of the treatment. There is no full agreement on the ideal calcium concentration in dialysis fluids, since this choice is also mostly influenced by the concomitant medical therapy. However, there is some consensus in suggesting a lower calcium concentration in standard hemodialysis (HD) treatment (1.25-1.50 mmol/l) than in dialysis treatments characterized by high convective transport. In peritoneal dialysis, calcium balance is affected by its blood dialysate concentration difference and dialysate glucose concentration, with ideal calcium concentration probably being >1.25 and <1.75 (which are the most commonly used concentrations). Phosphate dialysis balance is also a critical and challenging problem, since the possibility to reach an always desired null or possibly negative balance relies on this. Even though some increased phosphate removal can be obtained with the highest efficiency techniques, such as hemodiafiltration as compared with traditional HD, the most impacting factor still remains the duration of dialysis treatment. However, some experimental attempts at increasing phosphate removal independently of increasing dialysis duration are mentioned. PMID- 19295200 TI - Dietary and pharmacological control of calcium and phosphate metabolism in dialysis patients. AB - Chronic kidney disease-mineral and bone disorder is a new term defining a complex syndrome which underlines the need of a systemic approach to disturbances of calcium and phosphate metabolism in patients with renal failure. In recent years, the availability of new phosphorus binders and the appearance of new selective vitamin D receptor activators and calcimimetics have increased our current armamentarium and have changed previous paradigms. All these drugs can be used in combination, acting in distinct yet complementary pathways, with a resultant improvement in their individual clinical profile and reduction in secondary effects, while enhancing the achievement of clinical guideline targets. On the other hand, we should be aware that treatment costs are increasing and most of our knowledge is opinion-based. In this article, we shall consider rational recommendations on the control of calcium, phosphorus and parathyroid hormone while awaiting new evidence. We shall also briefly review some important related issues such as vascular calcification, adynamic bone disease, osteoporosis and the need of parathyroidectomy. Future guidelines may modify current recommendations, but we believe that the lack of an absolute evidence is not equivalent to the lack of awareness of the important problem which chronic kidney disease-mineral and bone disorder represents. PMID- 19295201 TI - Frequency of sustained intracranial pressure elevation during treatment of severe intraventricular hemorrhage. AB - BACKGROUND: Elevated intracranial pressure (ICP) is an important marker of neurological deterioration. The occurrence and significance of elevated ICP and low cerebral perfusion pressure (CPP) in aggressively treated spontaneous intraventricular hemorrhage (IVH) are not defined. METHODS: We performed a secondary longitudinal exploratory data analysis of a randomized multicenter trial of urokinase (UK) versus placebo (Pcb) as a treatment for IVH. Eleven IVH patients who required an external ventricular drain (EVD) were randomized to receive either intraventricular UK or Pcb every 12 h until clinical response permitted EVD removal. ICP and CPP were recorded every 4 or 6 h, as well as before and 1 h after EVD closure for administration of study agent. ICP, CPP and the proportion of ICP readings above 20, 30, 40 and 50 mm Hg were analyzed. RESULTS: Six UK and 5 Pcb patients aged 39-74 years (mean +/- standard deviation; 53 +/- 11 years) were enrolled. Initial ICP ranged from 0 to 38 mm Hg (10.9 +/- 11.0), initial CPP from 65 to 133 mm Hg (100.5 +/- 17.7). We recorded 472 ICP readings over the entire monitoring period. Of these 65 (14%) were >20 mm Hg, 23 (5%) >30 mm Hg, 9 (2%) >40 mm Hg and 3 (<1%) >50 mm Hg. Only 2 of 141 intraventricular injections of study agent with EVD closure were not tolerated and required reopening of the EVD. CONCLUSIONS: In the intensive care unit, initial ICP measured with an EVD was uncommonly elevated (1/11 patients) in this group of severe IVH patients despite acute obstructive hydrocephalus. Frequent monitoring reveals ICP elevation >20 mm Hg in 14% of observations during use of EVD. ICP elevation, though it can occur, is not routinely associated with EVD closure for thrombolytic treatment with UK. PMID- 19295202 TI - Intraventricular hemorrhage: harmful effect is not from the pressure alone. PMID- 19295203 TI - Hemifacial spasm and pontine compression caused by a giant vertebrobasilar dolichoectasia. PMID- 19295204 TI - Limitation of brachial-ankle pulse wave velocity in assessing the risk of stroke: importance of instantaneous blood pressure. AB - BACKGROUND AND OBJECTIVE: The carotid-femoral pulse wave velocity is a well established index of aortic stiffness, and the brachial-ankle pulse wave velocity (baPWV) has recently been developed as a new method for evaluating arterial stiffness. However, whether the baPWV is also useful in evaluating the risk of stroke is currently controversial. Moreover, it is also unknown which types of stroke and neuroradiological findings are associated with increased arterial stiffness. We investigated the association between the baPWV and risk of stroke using a case-control design. METHODS: This study enrolled 223 stroke patients (aged 65.6 +/- 10.3 years, mean +/- SD) and the same number of age- and sex matched controls. All of the subjects in this study underwent a structured interview and had their vascular risk factors assessed. We reviewed brain magnetic resonance imaging and magnetic resonance angiography data of stroke patients to assessthe severity of white-matter hyperintensity lesions, number of cerebral microbleeds, and the stenosis of both intracranial and extracranial arteries. The baPWV was measured noninvasively by an oscillometric method. RESULTS: The baPWV was most affected by the instantaneous systolic blood pressure, and was significantly higher in stroke patients than control subjects (1,721 +/- 395 cm/s vs. 1,657 +/- 366 cm/s, p = 0.03). However, the baPWV did not differ significantly with a specific type of stroke or subtype of ischemic stroke. Multiple regression analysis revealed that the baPWV was not independently associated with increased risk of stroke, or the severity of white matter hyperintensity lesions or cerebral microbleeds. CONCLUSION: Although the baPWV was significantly higher in stroke patients than control subjects, it was not independently associated with increased risk of stroke or specific subtypes of ischemic stroke or neuroradiological findings. PMID- 19295205 TI - Is the NIHSS certification process too lenient? AB - BACKGROUND AND PURPOSE: The National Institutes of Health Stroke Scale (NIHSS) is a widely used measure of neurological function in clinical trials and patient assessment; inter-rater scoring variability could impact communications and trial power. The manner in which the rater certification test is scored yields multiple correct answers that have changed over time. We examined the range of possible total NIHSS scores from answers given in certification tests by over 7,000 individual raters who were certified. METHODS: We analyzed the results of all raters who completed one of two standard multiple-patient videotaped certification examinations between 1998 and 2004. The range for the correct score, calculated using NIHSS 'correct answers', was determined for each patient. The distribution of scores derived from those who passed the certification test then was examined. RESULTS: A total of 6,268 raters scored 5 patients on Test 1; 1,240 scored 6 patients on Test 2. Using a National Stroke Association (NSA) answer key, we found that correct total scores ranged from 2 correct scores to as many as 12 different correct total scores. Among raters who achieved a passing score and were therefore qualified to administer the NIHSS, score distributions were even wider, with 1 certification patient receiving 18 different correct total scores. CONCLUSIONS: Allowing multiple acceptable answers for questions on the NIHSS certification test introduces scoring variability. It seems reasonable to assume that the wider the range of acceptable answers in the certification test, the greater the variability in the performance of the test in trials and clinical practice by certified examiners. Greater consistency may be achieved by deriving a set of 'best' answers through expert consensus on all questions where this is possible, then teaching raters how to derive these answers using a required interactive training module. PMID- 19295206 TI - High resolution wall and lumen MRI of the middle cerebral arteries at 3 tesla. AB - BACKGROUND: Although black-blood MRI (BB-MRI) can identify plaques in the cervical carotid arteries, this modality has not been applied in intracranial arteries. We imaged the lumina and walls of stenotic middle cerebral arteries (MCAs) in symptomatic and asymptomatic patients using high-resolution BB-MRI, in order to characterize vulnerable plaques and to determine the diagnostic accuracy of BB-MRI in MCA stenosis. METHODS: Multicontrast (T(1), T(2) and proton density) weighted BB-MRIs were acquired in 15 patients with MCA stenosis and in 2 volunteers. Each MCA was classified into one of three groups based on MR angiographic findings and symptoms: normal, symptomatic stenosis, or asymptomatic stenosis. The plaque signal intensity was interpreted and the total wall thickness was measured at the most stenotic segment. These values were then compared between asymptomatic and symptomatic MCAs using t test. For assessment of lumen imaging, the MCA stenosis graded on BB-MR images was compared with that graded on conventional angiography (digital subtraction angiography). RESULTS: Twenty-eight MCAs were evaluated (normal MCAs: 12, symptomatic stenoses: 7, and asymptomatic stenoses: 9). T(1)- and/or T(2)-hyperintense foci were demonstrated more frequently within the plaques of symptomatic stenoses than within the plaques of asymptomatic stenoses (57.1 vs. 22%). The total wall thickness in the symptomatic stenoses was significantly higher than that seen in the asymptomatic stenoses. The stenosis grade for the BB-MRI was significantly correlated with the digital subtraction angiography grade. CONCLUSION: High-resolution, multicontrast weighted BB-MRI has the potential to characterize atherosclerotic plaques in the MCA and may be a useful modality for evaluating the degree of stenosis. PMID- 19295207 TI - Cerebral embolism during carotid artery stenting: role of carotid plaque echolucency. AB - BACKGROUND: Carotid artery stenting (CAS) is associated with the risk of intraprocedural stroke. A better understanding of specific risk factors could help to improve the procedure and to reduce the overall risk of CAS. We addressed the role of carotid plaque echolucency as potential risk factor for cerebral embolism during CAS. METHODS: We prospectively evaluated carotid plaque echolucency by use of a computer-assisted measure of echogenicity, the gray scale median (GSM), in 31 consecutive patients with symptomatic high-grade carotid stenosis that were scheduled to undergo CAS. Dual-frequency transcranial Doppler ultrasound was used to detect solid cerebral microemboli during CAS. RESULTS: 27 of the 31 patients met all inclusion/exclusion criteria.Solid cerebral microemboli were detected during 17 of 27 CAS procedures. The GSM of the target plaques was lower in subjects with intraprocedural embolism (37.9 +/- 20.8) than in those without (58.2 +/- 25.7) (p = 0.040). A receiver-operating characteristic analysis showed that the GSM that gave the greatest separation between plaques with a higher and a lower probability of intraprocedural embolism was 50: the proportion of subjects with intraprocedural embolism was 85% in CAS of echolucent plaques (GSM <50) and 42% in CAS of echogenic plaques (GSM > or =50) (p = 0.031). CONCLUSIONS: CAS of both echolucent and echogenic carotid plaques may be associated with cerebral embolism, particularly CAS of echolucent plaques. Plaque echolucency alone does not reliably identify patients at particularly high risk of intraprocedural embolism, but should be considered as one of a broad panel of risk factors of CAS. PMID- 19295208 TI - Diagnostic evaluation for patients with ischemic stroke: are there sex differences? AB - BACKGROUND AND PURPOSE: Differences in the management of women and men with acute coronary symptoms are well documented, but relatively little is known about practices for patients with ischemic stroke. We sought to determine whether there are sex-associated differences in the utilization of diagnostic tests for ischemic stroke patients treated at academic hospitals in the United States. METHODS: Medical records were abstracted for consecutive ischemic stroke patients admitted to 32 US academic medical centers from January through June, 2004, as part of the University HealthSystem Consortium Ischemic Stroke Benchmarking Project. We compared the utilization rates of diagnostic tests including neuroimaging (CT or MRI), electrocardiogram (ECG), ultrasound of the carotid arteries, and echocardiography (transthoracic or transesophageal) for women and men. Multivariate logistic regression was used to test for sex differences with adjustment for potential confounders. RESULTS: The study included 1,256 ischemic stroke patients (611 women; 645 men; mean age 66.6 +/- 14.6 years; 56% white). There were no differences between women and men in the use of neuroimaging (odds ratio, OR = 1.37; 95% confidence interval, CI = 0.58-3.24), ECG (OR = 1.00, 95% CI = 0.70-1.44), carotid artery ultrasound (OR = 0.93, 95% CI = 0.72-1.21) or echocardiography (OR = 0.70, 95% CI = 0.70-1.22). The results were similar after covariate adjustment. CONCLUSIONS: Women and men admitted to US academic hospitals receive comparable diagnostic evaluations, even after adjusting for sociodemographic and clinical factors. PMID- 19295209 TI - Preparation with oral tablets of sodium phosphate (Visiclear) is suitable for colonoscopy but not for colonoscopy with upper endoscopy. PMID- 19295210 TI - Early Helicobacter pylori eradication restores sonic hedgehog expression in the gastric mucosa of Mongolian gerbils. AB - BACKGROUND AND AIM: Sonic hedgehog (Shh) is a morphogen involved in the homeostasis of the gastric fundic glands. Alterations of gastric mucosal Shh expression after eradication of Helicobacter pylori were examined. METHOD: Mongolian gerbils were inoculated with H. pylori at the age of 5 weeks. H. pylori eradication was then carried out at 12, 24 or 48 weeks after the inoculation, and the gerbils were examined at 10 weeks after the eradication. Gastric inflammation was evaluated by the tissue myeloperoxidase activity and the histological scoring. Immunohistochemistry and in situ hybridization were performed for determining the Shh expression. RESULTS: Significant decrease of the myeloperoxidase activity and scores for acute and chronic inflammation as well as atrophy were observed in the H. pylori-eradicated gerbils as compared with the findings in the non-H. pylori-eradicated gerbils. Significant increase of the horizontal length of the area positive for Shh expression was noted in the H. pylori-eradicated gerbils as compared with that in the non-H. pylori-eradicated gerbils. Earlier eradication promoted better restoration of Shh expression. 50% of the animals of the 24-week eradication group and all animals in the 48-week eradication group exhibited heterotopic proliferative glands. In the animals showing heterotopic proliferative glands, the front line of Shh regeneration was cut off at the point of development of heterotopic proliferative glands. CONCLUSION: H. pylori-associated deregulation of Shh expression that could be linked to gastric atrophy and the associated preneoplastic transformation appears to be reversible with early H. pylori eradication. PMID- 19295211 TI - Normal values of 24-hour combined esophageal multichannel intraluminal impedance and pH monitoring in the Chinese population. AB - BACKGROUND: There is a lack of data on combined esophageal multichannel intraluminal impedance and pH (MII-pH) monitoring in the Chinese population. GOALS: To define the normal range and compare the parameters to those of the Western population. STUDY: Healthy volunteers were recruited and underwent 24 hour ambulatory combined MII-pH monitoring. Gastroesophageal reflux episodes were detected and characterized by pH (acidic, weakly acidic and weakly alkaline reflux) and composition (liquid, mixed and gas reflux). RESULTS: Seventy healthy volunteers (male:female 33:37) were recruited. The median and 95th percentile number of the total reflux episodes over 24 h in the Chinese population was 40 and 75, respectively, whereas the corresponding numbers of acidic, weakly acidic and weakly alkaline reflux episodes were 22 and 54, 16 and 40, and 0 and 4, respectively. Among all reflux episodes 53.3% were acidic, 44.7% were weakly acidic, and 2% weakly alkaline. More than half (52.4%) of the reflux episodes were mixed, 37.2% were liquid, and 10.4% were gas. A total of 26.6% of reflux episodes reached 15 cm above the lower esophageal sphincter. Male gender was associated with an increased number of acid, liquid, mixed and proximal reflux episodes (p < 0.05). CONCLUSIONS: This study provides normal values for the Chinese population. Differences between genders in impedance parameters required caution during data interpretation. The number of reflux episodes in the Chinese population was similar to that in the Western population. PMID- 19295212 TI - Cognitive impairment in spinocerebellar degeneration. AB - It has been reported that patients with spinocerebellar degenerations (SCDs) have cognitive dysfunction as well as limb and truncal ataxia, dysarthria and dysphagia. We review cognitive dysfunction in common types of SCD, including spinocerebellar ataxia types 1, 2, 3, 6, and 17, dentatorubral-pallidoluysian atrophy, Friedreich's ataxia, and multiple system atrophy. There are few studies that address cognitive function in SCD. Although there are few comparison studies among the various SCDs, cognitive dysfunction may be more common and severe in spinocerebellar ataxia type 17 and dentatorubral-pallidoluysian atrophy. While cognitive dysfunction in SCD appears to represent frontal dysfunction, the mechanisms of cognitive dysfunction have not been directly clarified. Nevertheless, various lesions, including those in the cerebrocerebellar circuitry, cortico-striatal-thalamocortical circuitry, and the frontal lobe, may influence cognitive function to various degrees for each disease. PMID- 19295213 TI - Amusia and musical functioning. AB - Music, as language, is a universal and specific trait to humans; it is a complex ability with characteristics that are unique compared to other cognitive abilities. Nevertheless, several issues are still open to debate, such as, for example, whether music is a faculty that is independent from the rest of the cognitive system, and whether musical skills are mediated by a single mechanism or by a combination of processes that are independent from one another. Moreover, the anatomical correlations of music have yet to be clarified. The goal of this review is to illustrate the current condition of the neuropsychology of music and to describe different approaches to the study of the musical functions. Hereby, we will describe the neuropsychological findings, suggesting that music is a special function carried out by different and dedicated processes that are probably subserved by different anatomical regions of the brain. Moreover, we will review the evidence obtained by working with brain-damaged patients suffering from music agnosia, a selective impairment in music recognition. PMID- 19295214 TI - Patients' stratification and correlation of brain magnetic resonance imaging parameters with disability progression in multiple sclerosis. AB - AIMS: The objective was to correlate the change in the lesion load (LL) and brain atrophy in patients with multiple sclerosis (MS) with progression of clinical disability, represented by the Expanded Disability Status Scale (EDSS), and to test if stratification of patients according to magnetic resonance imaging (MRI) criteria can increase the predictive ability of MRI for MS clinical development. METHODS: 181 patients with clinically definite relapsing-remitting MS underwent MRI for a period of up to 5 years. Grouping of patients according to the LL value at the study entry revealed a substantial increase in the Spearman rank correlation coefficient R. RESULTS: For the low LL cohort of patients, we found a statistically significant correlation (R up to -0.71 with p < 0.01) of a later increase in the EDSS score (years 4 and 5) with increased brain atrophy in the first 2 years. For the high LL group, we found a statistically significant correlation (R up to 0.72 with p < 0.01) of a later increase in the EDSS score (years 4 and 5) with an increase in the LL value in the first year. CONCLUSIONS: We conclude that stratification of patients according to the MRI criterion (LL) can increase the predictive ability of MRI. PMID- 19295215 TI - Frequent premature atrial contractions in stroke of undetermined etiology. AB - BACKGROUND: The cause of ischemic stroke is undetermined in 15-40% of patients. We studied the association between frequent premature atrial contractions (PAC) and stroke of undetermined etiology. METHODS: We retrospectively studied ischemic stroke patients who were consecutively admitted to our department and underwent 24-hour Holter ECG recording including 163 noncardioembolic stroke patients (group A), 24 stroke patients of undetermined etiology (group B), and 37 cardioembolic stroke patients with prior-diagnosed paroxysmal atrial fibrillation (group C). The number of PAC per 24 h and the prevalence of patients with frequent PAC (200 or more per 24 h) were compared between the groups. RESULTS: The number of PAC was significantly larger in groups B and C than in group A. The proportion of patients with frequent PAC was larger in group B (13/24, 54%) and in group C (18/37, 48%) than in group A (32/163, 20%). CONCLUSION: Our data suggest that frequent PAC should be regarded as a masked type of paroxysmal atrial fibrillation and should be included in one of the causes of cardioembolic stroke. PMID- 19295216 TI - Encephalitis after influenza in Sweden 1987-1998: a rare complication of a common infection. AB - The aim of this study was to investigate the incidence of influenza-related encephalitis in Sweden during 11.5 years. Studies from Japan report an increased incidence of influenza-related encephalitis/encephalopathy. Few other studies are available. We conducted a retrospective register-based study on the Swedish National Inpatient Register, which covers all Swedish hospitals. In 1987-1998, a total number of 14,250 hospitalized individuals had an influenza diagnosis (population incidence: 137 per million person-years). In-hospital mortality was 4.1%. Using three different approaches, only 21 cases of influenza-related encephalitis were found, corresponding to a rate of 1.5 per 1,000 hospitalized persons with an influenza diagnosis (population incidence 0.21 per million person years). We conclude that encephalitis following influenza occurs rarely, or is an infrequently recognized, diagnosed or reported complication. The cases we studied in detail have all recovered without sequels. PMID- 19295217 TI - Clinical identification of dysarthria types among neurologists, residents in neurology and speech therapists. AB - BACKGROUND: Classification of dysarthria types comprises flaccid, spastic, ataxic, hypo- and hyperkinetic and mixed dysarthria. This study focussed on the ability of neurologists to clinically identify the correct type of dysarthria in neurological patients. METHODS: Eighteen patients with dysarthria and 4 healthy controls were enrolled in the study. The gold standard for dysarthria type was the underlying neurological disease. Recordings of a standard reading passage and free speech were made. Raters were neurologists, residents in neurology and speech therapists, whose scores were compared. RESULTS: Neurologists correctly identified 40% of the recordings, residents 41%, and speech therapists 37%. Interrater agreement was fair among all 3 groups; intrarater agreement was fair to moderate. CONCLUSION: This study suggests that neurologists should be aware of the unreliability of identifying the dysarthria type without the use of additional validated instruments or rating scales in a clinical setting. PMID- 19295218 TI - Clinical features, neuroimaging and treatment of spontaneous intracranial hypotension and magnetic resonance imaging evidence of blind epidural blood patch. AB - BACKGROUND AND PURPOSE: Spontaneous intracranial hypotension (SIH) is an uncommon, but not rare, cause of headache. We analyzed a series of patients with SIH and attempted to establish a clinical procedure. METHODS: We retrospectively reviewed 11 patients with SIH who were admitted between January 2004 and May 2007. Data recorded from patients included basic data, clinical symptoms, neurological imaging studies and treatment. RESULTS: Orthostatic headache was the most common symptom, but there were still 2 patients without orthostatic headache. Diffuse pachymeningeal enhancement was the most common finding in brain MRI study and engorgement of the spinal epidural venous plexus was the most common finding in spinal MRI study. Eight of our patients received epidural blood patches in the lumbar area and 6 of these were symptom-free within 2 weeks. Two patients received spinal MRI immediately after administering the epidural blood patch, and this revealed that most of the blood had spread to the upper cervical area from the lumbar injection. CONCLUSION: A blind epidural blood patch from the lumbar area is an acceptable procedure even if the area of leakage is unknown. A reasonable clinical procedure for the patients of SIH may minimize the rate of repeat puncture. PMID- 19295219 TI - Isolated insular infarction following successful intravenous thrombolysis of middle cerebral artery strokes. PMID- 19295220 TI - Marie-Jean-Pierre Flourens (1794-1867) and cortical localization. AB - The child prodigy Marie-Jean-Pierre Flourens received his medical degree at Montpellier when aged 19. As a young promising physician Flourens was asked to investigate Gall's controversial views on cerebral localization. To test Gall's assertions, Flourens developed ablation as a procedure to explore the workings of the brain. By removing anatomically defined areas of the brain of an animal and watching its behaviour, he thought he might localize certain functions. Flourens did not favour the idea of cerebral localization and concluded that the brain functioned as a whole and thus arose the concept of 'cerebral equipotentiality'. This culminated in his 1824 Recherches experimentales sur les proprietes et les fonctions du systeme nerveux. His techniques were, however, crude and imperfect, and his experiments were mainly on birds. Much criticism and debate ensued. A gifted man, Flourens also advanced the physiology of the vestibular apparatus and described the anaesthetic properties of ether. PMID- 19295221 TI - Delayed postanoxic encephalopathy with serial MRI and PET studies. PMID- 19295222 TI - Effect of supplemental oxygen versus dobutamine administration on liver oxygen tension in dPP-guided normovolemic pigs. AB - BACKGROUND: Difference in pulse pressure (dPP) confirms adequate intravascular filling as a prerequisite for tissue perfusion. We hypothesized that both oxygen and dobutamine increase liver tissue oxygen tension (ptO(2)). METHODS: Eight anesthetized pigs received dPP-guided fluid management. Hepatic pO(2) was measured with Clark-type electrodes placed subcapsularly, and on the liver surface. Pigs received: (1) supplemental oxygen (F(i)O(2) 1.0); (2) dobutamine 2.5 microg/kg/min, and (3) dobutamine 5 microg/kg/min. Data were analyzed using repeated-measures ANOVA followed by a Tukey post-test for multiple comparisons. ptO(2 )measured subcapsularly and at the liver surface were compared using the Bland-Altman plot. RESULTS: Variation in F(i)O(2) changed local hepatic tissue ptO(2) [subcapsular measurement: 39 +/- 12 (F(i)O(2) 0.3), 89 +/- 35 mm Hg (F(i)O(2) 1.0, p = 0.01 vs. F(i)O(2) 0.3), 44 +/- 10 mm Hg (F(i)O(2) 0.3, p = 0.05 vs. F(i)O(2) 1.0); surface measurement: 52 +/- 35 (F(i)O(2) 0.3), 112 +/- 24 mm Hg (F(i)O(2) 1.0, p = 0.001 vs. F(i)O(2) 0.3), 54 +/- 24 mm Hg (F(i)O(2) 0.3, p = 0.001 vs. F(i)O(2) 1.0)]. Surface measurements were widely scattered compared to subcapsular measurements (bias: -15 mm Hg, precision: 76.3 mm Hg). Dobutamine did not affect hepatic oxygenation. CONCLUSION: Supplemental oxygen increased hepatic tissue pO(2) while dobutamine did not. Although less invasive, the use of surface measurements is discouraged. PMID- 19295223 TI - [Complementarity: supplementarity and oppositeness]. PMID- 19295224 TI - [Laser needle acupuncture in women with dysmenorrhoea: a randomised controlled double blind pilot trial]. AB - BACKGROUND: Dysmenorrhoea is a common gynaecological disorder.It is treated symptomatically if it is not caused by any underlying disease. Acupuncture is successfully used for menstrual disorders but is associated with certain side effects such as pain. Laser needle acupuncture is a new, non-invasive procedure that might mimic the effects of acupuncture. OBJECTIVE: The aim of this study is to investigate whether the effect of active laser needle acupuncture is superior to the effect of placebo laser needle acupuncture in women with dysmenorrhoea. PATIENTS AND METHODS: Women aged 18-50 years and with a minimum menstrual pain of 4 out of 10 points on a visual analogue scale (VAS) were included in this randomised double blind trial and treated with laser needle acupuncture (Laserneedle Computer-System). Every woman was treated with the same 8 acupuncture points (SP6, LV3 and LI4 bilateral; CV3 and ST36 on the right) in 8 sessions of 20 min each, over a period of 3 menstrual cycles. The primary endpoint was successful pain reduction, defined as a 50% reduction of the mean menstrual pain between baseline and end of the study (calculated from those 2 menstrual days with the worst pain levels on the VAS). We compared real laser acupuncture with placebo laser acupuncture (no laser activity). RESULTS: 48 women, mean age 29.6 +/- 7.5 years, were included in the study (30 women in the placebo group, 18 in the verum group). In the real acupuncture group, the success rate was 16.7% (3/18), in the placebo group 20% (6/30). CONCLUSIONS: We could not find a significant advantage of laser needle acupuncture compared to placebo laser needle acupuncture treatment. Further investigations are recommended because laser needle treatment is non-invasive and allows double blind acupuncture studies. PMID- 19295225 TI - Efficacy of homeopathically potentized antimony on blood coagulation. A randomized placebo controlled crossover trial. AB - BACKGROUND: Homeopathically potentized antimony 6x is traditionally used in anthroposophic medicine for an alleged pro-coagulatory effect in bleeding disorders. However, the scientific evidence base is yet insufficient. Results of a previous in vitro study suggested a slight increase of maximal clot firmness (MCF) and a tendency towards a shorter clotting time (CT). The objective of this study was to investigate the pro-coagulatory effects of antimony in vivo, and possible unexpected or adverse events. PARTICIPANTS AND METHODS: A randomized placebo controlled double blind crossover study was carried out in 30 healthy volunteers (15 males, 15 females). Each participant received intravenously 10 ml of antimony 6x and placebo in a randomized order at an interval of 1 month. Thrombelastography (TEG) was carried out immediately before and 30 and 60 min after the injection. RESULTS: Statistically significant pro-coagulatory effects were observed 30 min after injection for CT in men (p = 0.0306), and for MCF in men and women combined (p = 0.0476). The effect of antimony was significantly larger on test day 1 than on test day 2, whereas the effect of placebo was similar on both test days. No unexpected adverse or adverse events causally related to antimony were observed. CONCLUSION: This study adds evidence to the hypothesis that homeopathically potentized antimony may be efficacious in vivo. The consistency of the results with previous in vitro results indicates an effect on MCF and CT. The in vivo application of antimony 6x is safe. PMID- 19295226 TI - BCQ+: a body constitution questionnaire to assess Yang-Xu. Part II: Evaluation of reliability and validity. AB - INTRODUCTION: Integrated medicine has become an important issue in this century. In order to bridge the gap between traditional Chinese medicine (TCM) and modern medicine, we have established a first final version of a Yang-Xu constitution questionnaire, the BCQ+, in a previous study. The objective of the present investigation is to assess the reliability and validity of the BCQ+. PARTICIPANTS AND METHODS: A total of 1,310 eligible participants, aged between 20-60, no disease onset in the past month, who received health examination in teaching hospitals were recruited. The BCQ+ was developed and psychometrically evaluated to ascertain and validate the factor structure of the BCQ+. Furthermore, the reliability and validity of the BCQ+ were examined. RESULTS: 19 items from the BCQ+ were developed. Exploratory factor analysis identified 5 primary domains that represent the Yang-Xu constitution and correspond to the diminishing energy levels of 5 different body areas: head, chest, limbs, abdominal cavity, and body surface. Cronbach's alpha ranged between 0.55-0.88 and the intraclass correlation coefficients were >0.7 for most of the 19 questions. The diagnoses of Yang-Xu by the BCQ+ and an experienced TCM doctor were consistent. Systolic blood pressure (SBP) was used as an indicator of concurrent validity of the BCQ+, and it was found that higher Yang-Xu scores were associated with a lower SBP. CONCLUSIONS: Five domains with 19 items in the BCQ+ were developed. The BCQ+ is a valid and potentially useful tool for evaluating the Yang-Xu constitution in clinical practice and research. PMID- 19295227 TI - [Art therapy for cancer patients in outpatient care. Psychological distress and coping of the participants]. AB - BACKGROUND: Various types of art therapy increasingly gain importance in psycho oncology. The aim of this article is to determine whether art therapy may help decrease psychological distress and increase coping skills in cancer patients. PATIENTS AND METHODS: An art therapy course for use in psycho-oncological care for outpatients was developed and implemented in a prospective observation study of the Department of Social Medicine,Leipzig University. Participants' levels of psychological distress (HADS) as well as their coping skills (TSK) were quantitatively evaluated before (t1) and after (t2) the intervention. RESULTS: After completion of the course mean anxiety of the participants(n = 18) had significantly decreased from 11.06 to 9.33 (p < 0.04). On average, however, participants reported more anxiety both before and after the intervention than a representative comparison group of n = 48 cancer patients (anxiety: mean= 5.51). Regarding depression no significant change could be observed after completion of the intervention (t1 = 6.94, t2 =6.22; p < 0.32). Also in relation to the comparison group (mean= 5.24) no significant difference in depression could be detected (t1: p < 0.066, t2: p < 0.519). No significant change in participants' coping skills could be observed. CONCLUSIONS: The level of psychological distress among cancer patients in outpatient care is above average. This underlines the need for psycho-oncological aftercare services. Art therapy interventions can make an important contribution to the psychological well-being of cancer patients. PMID- 19295228 TI - Phyllanthus urinaria increases apoptosis and reduces telomerase activity in human nasopharyngeal carcinoma cells. AB - BACKGROUND: This study was designed to obtain the chemical fingerprint and to investigate the effect of Phyllanthus urinaria on telomerase activity and apoptotic pathways in the human nasopharyngeal carcinoma cell line (NPC-BM1). MATERIALS AND METHODS: The polyphenol compounds in P. urinaria were investigated by HPLC/MS. Cell viability with the treatment of P. urinaria, gallic acid, ellagic acid, quercetin and cisplatin was detected by MTT assay. TUNEL assay, DNA fragmentation analysis and caspase3 activity were used to confirm apoptotic changes. Telomerase activity was determined using the TRAP assay. RNA isolation and RT-PCR were used to analyze the related genes expression. All experiments on treatments with P. urinaria from 0-3 mg/ml were carried out for 24 h. RESULTS: 5 major compounds including gallic acid, brevifolin carboxylic acid, corilagin, phyllanthusiin C and ellagic acid were identified as a plant fingerprint by HPLC/MS. With the MTT assay, we demonstrated that P. urinaria, gallic acid and ellagic acid reduce cell viability. The apoptosis features showed DNA fragmentation and increased caspase-3 activity associated with the down regulation of Bcl-2, but not of Bax, p53, and PCNA (proliferating cell nuclear antigen) in P. urinaria-treated NPC-BM1 cells. Furthermore, treatment of NPC-BM1 cells led to an inhibition of hTERT (human telomerase reverse transcriptase), hTP1 (human telomerase-associated protein 1) and c-myc mRNA expression and to decreased telomerase activity. CONCLUSION: This study suggests that P. urinaria induces the death of NPC-BM1 cells in vitro through the induction of apoptosis and inhibited telomerase activity. PMID- 19295229 TI - Mangrove plant extracts: radical scavenging activity and the battle against food borne pathogens. AB - AIMS: The present study was carried out to evaluate the radical scavenging and antibacterial activity of methanolic leaf extracts of mangrove plants. 8 mangrove plants, which are used as folk medicine in various tropical countries, were collected in Pichavaram and Thondi, Tamil Nadu, India. METHODS: The antioxidant properties of methanolic leaf extracts were assessed under in vitro conditions using different antioxidant tests, including DPPH, nitric oxide, hydrogen peroxide, hydroxyl radical scavenging assay, reducing power, ferrous ion chelating, and lipid peroxidation inhibition assay. RESULTS AND CONCLUSION: Of the 8 mangrove plants screened, Rhizophora mucronata (100 microg/ml) showed significantly higher (p < 0.05) activities for all antioxidant assays, and its IC50 values were 43.17, 116, 60.06, and 46.76 microg/ml for DPPH, hydrogen peroxide, nitric oxide, and hydroxyl radical scavenging activity, respectively. Butylated hydroxytoluene, ascorbic acid and alpha-tocopherol were used as positive controls. Methanolic extracts of R. mucronata were also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals in a Fenton's reaction mixture. Moreover, the total phenolic content of R. mucronata (720.79 mg/gm of dry leaves) was very high and a strong positive correlation was observed between its phenolic content and its antioxidant activity. Methanolic leaf extracts of mangrove plants exhibited no antibacterial activity against seven food-borne pathogens studied. PMID- 19295230 TI - [Favourable course of persisting malignant ascites]. AB - Malignant ascites is a frequent complication in oncological diseases. There are no standard therapies for any primary tumour. We report the case of a woman, aged 49 years at the time of primary diagnosis, who suffered from recurrent ascites resulting from liver metastasis of breast cancer. Based on the literature and former experience of our department, mistletoe extract was repeatedly applied intraperitoneally at the occasion of decompressive punctures. The further course of the disease suggests a significant role of mistletoe in achieved symptom control, which also resulted in a considerable improvement in quality of life. The mistletoe solution was well tolerated. Relevant mechanisms of action in addition to the well-known immunomodulating properties of mistletoe could be direct cytotoxic and adjuvant effects to the concomitantly administered chemotherapy of carboplatin/paclitaxel. PMID- 19295231 TI - Watermelon contains 92% water but it also contains allergens! PMID- 19295232 TI - Identification of major allergens in watermelon. AB - BACKGROUND: Watermelon is a worldwide consumed Cucurbitaceae fruit that can elicit allergic reactions. However, the major allergens of watermelon are not known. The aim of this study is to identify and characterize major allergens in watermelon. METHODS: Twenty-three patients allergic to watermelon took part in the study. The diagnosis was based on a history of symptoms and positive skin prick-prick tests to watermelon, confirmed by positive open oral challenge testing to watermelon pulp. Allergenic components were detected by SDS-PAGE and immunoblotting. Molecular characterization of IgE-binding bands was performed by N-terminal amino acid sequencing and mass spectrometry. Allergens were purified combining several chromatographic steps. RESULTS: Several IgE binding bands (8 120 kDa) were detected in watermelon extract. Three major allergens were identified as malate dehydrogenase (36 kDa), triose phosphate isomerase (28 kDa) and profilin (13 kDa). Purified allergens individually inhibited IgE binding to the whole watermelon extract. CONCLUSIONS: All in all these results indicate that malate dehydrogenase, triose phosphate isomerase and profilin are major allergens involved in watermelon allergy. PMID- 19295233 TI - Development of an adjuvant-free cashew nut allergy mouse model. AB - BACKGROUND: Cashew nut allergy is an emerging food allergy with a high risk of systemic anaphylaxis. Currently, an adjuvant-free animal model to study cashew nut allergy is not available. METHODS: BALB/c mice were exposed to cashew nut protein using a transdermal sensitization protocol that does not use adjuvant. Systemic IgE antibody response, systemic anaphylaxis to oral challenge and allergen-driven, spleen-cell, type-2 cytokine responses were studied. RESULTS: Transdermal exposure to cashew nut resulted in a significant dose-dependent allergic response. Oral challenge of sensitized mice with cashew resulted in severe signs of systemic anaphylaxis and a significant hypothermia. Spleen cell culture with cashew nut protein resulted in allergen-driven IL-4, IL-5 and IL-13 responses only in sensitized but not in saline control mice. CONCLUSIONS: These data demonstrate that (i) transdermal exposure to cashew nut protein elicits a robust IgE response leading to clinical sensitization of mice for systemic anaphylaxis to oral cashew nut challenge; (ii) cashew nut is a potent activator of type-2 cytokines, thus explaining the mechanism of cashew allergy, and (iii) this mouse model may be useful for further basic and preclinical studies on cashew nut allergy. PMID- 19295234 TI - Clinico-immunochemical studies on airborne Areca catechu L. Pollen, a probable risk factor in emergency asthma hospitalization from Eastern India. AB - BACKGROUND: The pollen grain of the Areca catechu L. tree is airborne and allergenic. This study aimed to know the role of this pollen as a source of aeroallergen with effect on emergency asthma hospitalization, to isolate its important allergic fraction and to check its cross-reaction with betel nut. METHODS: Areca pollen was monitored with a Burkard sampler. Determination of allergenic activities was studied by in vivo and in vitro analyses. Asthma hospitalization data were collected from two nearby hospitals. The pollen extract was fractionated by a combination of DEAE-Sephadex and Sephacryl S-200 column. The protein components were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cross-reactivity of Areca pollen and betel nut was shown by IgE enzyme-linked immunosorbent assay (ELISA) inhibition. RESULTS: The Areca pollen was perennially airborne. Skin test results of respiratory allergic patients showed 38.6% positivity. The detected aeroallergen spots in particle immunoblotting correlated significantly with airborne pollen count. Areca pollen showed a significant positive correlation with asthma hospitalization. There are 6 IgE-reactive protein components in the whole-pollen extract. IgE-reactive fraction 1 was resolved into 4 subfractions. Subfraction 1a showing IgE reactivity contained 3 protein components, among which 2 of 48 and 118 kDa were IgE reactive. The 48-kDa component was reported to be cross-reactive with other palm pollen types. In IgE ELISA inhibition, the betel nut extract showed 50% inhibition with about 110 ng/ml concentration. CONCLUSION: A. catechu pollen is a significant contributor to the aeroallergen load in India. Its partially purified IgE-reactive fraction may be useful in therapeutics. The betel nut extract showed remarkable cross-reactivity with Areca pollen. PMID- 19295235 TI - IL-16 variability and modulation by antiallergic drugs in a murine experimental allergic rhinitis model. AB - BACKGROUND: Interleukin-16 (IL-16) is a cytokine that induces selective migration of CD4+ cells and participates in inflammatory diseases including allergic rhinitis. Histamine and prostaglandin D(2) are important chemical mediators of allergic inflammation, and antiallergic drugs are commonly used for the treatment of allergic rhinitis. It remains unknown whether treatment with the drugs affects IL-16. OBJECTIVE: We evaluated the variability of IL-16 and the effects of the antiallergic drugs fexofenadine (40 mg/kg/day) and ramatroban (30 mg/kg/day) on IL-16 in an OVA-sensitized BALB/c murine experimental allergic rhinitis model. METHODS: We measured the expression level of IL-16 protein in the mouse nasal septal mucosa by immunohistochemistry, and the serum level of IL-16 by ELISA. Several other parameters associated with allergic rhinitis (nasal symptoms, OVA specific IgE, eosinophil and T cell infiltration) were also measured. RESULTS: Local and systemic expressions of IL-16 were significantly increased in OVA sensitized mice when compared to the nonsensitized group. Fexofenadine and ramatroban significantly inhibited the following OVA-induced allergic features when compared to the nontreated sensitized group: sneezing, nasal rubbing, eosinophil infiltration, IL-16 expressions in nasal tissue, and serum IL-16 level. Serum OVA-specific IgE and local T cell infiltration were reduced, but they did not reach significant values. CONCLUSIONS: These results suggest that IL 16 was both systemically and locally upregulated in the murine allergic rhinitis model and that IL-16 changed in parallel to allergic state by treatment with the drugs. PMID- 19295236 TI - Subcutaneous prostaglandin E(2) restrains airway mast cell activity in vivo and reduces lung eosinophilia and Th(2) cytokine overproduction in house dust mite sensitive mice. AB - BACKGROUND: Prostaglandin (PG) E(2) is thought to exert protective effects in the lungs. Accordingly, aerosolized PGE(2) prevents the experimentally induced airway response to allergen challenge in asthmatics. In vitro evidence indicating that functional PGE(2) receptors (EP) are expressed on human mast cells and that PGE(2) can alter cytokine production suggests that these phenomena may be involved in its beneficial effect in asthma. However, in vivo evidence is scarce. METHODS: We assessed the effects of exogenous PGE(2) and of the EP1/EP3 agonist sulprostone on the murine airway response to house dust mite (HDM) allergens, a model that accurately reproduces the spontaneous exposure of allergic asthma patients to aeroallergens. We also analyzed the in vivo impact of PGE(2) on production in the murine airway of mast cell protease (mMCP)-1, a specific marker of lung mast cell activity, and on local production of cytokines. RESULTS: Exogenous PGE(2), but not sulprostone, reduced eosinophilic infiltration in HDM sensitized mice by half and led to a strong reduction in airway Th(2) cytokine expression. These anti- inflammatory effects were accompanied in vivo by a substantial reduction in HDM-induced upregulation of airway mMCP-1. Neither PGE(2) nor sulprostone had any effect on airway hyperresponsiveness to methacholine. CONCLUSIONS: Our results indicate that the anti-inflammatory effect of PGE(2) can be reproduced in vivo in HDM-sensitized mice and suggest that this protective effect is dependent in vivo on inhibition of the allergen-triggered proinflammatory activity of bronchial mast cells. Finally, the effect of PGE(2) is linked to reduced upregulation of airway Th(2) cytokines. PMID- 19295237 TI - Therapeutic surfactants modulate the viability of eosinophils and induce inflammatory mediator release. AB - BACKGROUND: There is increasing interest in testing surfactant preparations for asthma therapy. Previously, Curosurf was demonstrated to increase inflammation in allergic asthmatics. So far, little is known about the immunomodulatory effects of therapeutic surfactants, in particular concerning the interaction of surfactant components with eosinophils as key effector cells of the allergic airway inflammation. The aim of the present study was to determine the effect of different therapeutic surfactants on cellular functions of eosinophils. METHODS: Eosinophils were isolated from peripheral blood of atopic volunteers and incubated with the natural animal-derived surfactants Curosurf or Alveofact or the synthetic recombinant human surfactant Venticute at different concentrations for up to 42 h. RESULTS: Curosurf and Venticute modulated the viability of eosinophils. While incubation with Curosurf increased the number of necrotic eosinophils after 1, 20 and 42 h, Venticute increased the number of apoptotic and necrotic cells after 1 h, but there were no differences compared with control cells at later time points. All surfactant preparations increased the levels of eosinophil cationic protein after 20 h and, in addition, Curosurf enhanced eosinophil cationic protein release after 42 h. The supernatant of eosinophils induced chemotaxis against autologous eosinophils, and the presence of Curosurf, but not Alveofact or Venticute, augmented the chemotactic effect. Chemotaxis was partly blocked by inhibition of eotaxin but not by inhibition of leukotrienes or platelet-activating factor. CONCLUSIONS: Therapeutic surfactants differ in their effects on eosinophil viability and the accompanying release of inflammatory mediators and chemotactic signals. Proinflammatory effects were most pronounced for the natural surfactant Curosurf. PMID- 19295238 TI - Association analysis of brain-derived neurotrophic factor gene polymorphisms in asthmatic families. AB - BACKGROUND: A role for neuronal modulation of inflammation and airway hyper responsiveness has been well described in asthma, and neurotrophins provide the link between inflammation and neuronal dysfunction. Brain-derived neurotrophic factor (BDNF) is an important mediator in this interaction. The aim of this study was to analyze the possible relationship between polymorphisms of the gene encoding BDNF and susceptibility to asthma. METHODS: 341 families with at least 2 siblings with asthma were genotyped for 4 BDNF polymorphisms (rs6265, rs2030324, rs988748 and rs7124442). RESULTS: Analysis by family-based association tests revealed no significant association between any polymorphisms analyzed and asthma susceptibility. Furthermore, BDNF polymorphism was not associated with asthma related phenotypes such as FEV(1) % predicted, bronchial hyper-responsiveness, IgE level, asthma and atopy severity scores. CONCLUSIONS: Our results suggest that genetic variation in the BDNF gene does not contribute significantly to asthma susceptibility or severity. PMID- 19295239 TI - Cedar and cypress pollinosis and allergic rhinitis: quality of life effects of early intervention with leukotriene receptor antagonists. AB - BACKGROUND: Allergic rhinitis involves inflammation of the nasal passages. The use of nasal steroids is generally very effective in providing significant symptom relief. However, compliance for their use is sometimes poor. METHODS: To examine the efficacy of early intervention (before pollen dispersal) with oral cysteinyl leukotriene receptor antagonists (LTRA) on pollinosis in patients with allergy to cedar and Japanese cypress pollens, groups of subjects were treated with LTRA or a placebo for 4 weeks at the beginning of the cedar pollen dispersal season. Subsequently, all patients received nasal steroid therapy concomitantly with LTRA throughout the remaining period of the pollen dispersal season. The effects of such early treatment with LTRA on pollinosis were investigated using symptom scores from an allergy diary and quality of life (QOL) scores. RESULTS: Sneezing and nasal congestion scores were significantly lower in the LTRA pretreated subjects than observed in the placebo-pretreated patients between weeks 4 and 6 and weeks 3 and 5, respectively. QOL scores improved significantly in all domains after concomitant therapy with nasal steroids. The percent improvement in the nasal congestion score after the concomitant therapy was significantly higher in the LTRA group (69%) than in the placebo group (41%). CONCLUSION: Significant differences observed in symptoms and in QOL effects between LTRA- and placebo-pretreated patients and the absence of major adverse effects noted in these studies suggest that early intervention with LTRA is beneficial and safe and should be considered in the management of pollinosis associated allergic rhinitis. PMID- 19295240 TI - Effect of enzymatically modified isoquercitrin, a flavonoid, on symptoms of Japanese cedar pollinosis: a randomized double-blind placebo-controlled trial. AB - BACKGROUND: Flavonoids exert antiallergic and antioxidant effects. We investigated the efficacy of enzymatically modified isoquercitrin (EMIQ), a flavonoid, to relieve symptoms of pollinosis. METHODS: In a parallel-group, double-blind placebo-controlled study design, 20 subjects with Japanese cedar pollinosis took two capsules daily of 100 mg EMIQ or a placebo for 8 weeks during the pollen season. Subjective symptoms and activities of daily living (ADL) scores were recorded every day, and the quality of life (QOL) score was obtained every 4 weeks. Blood sampling was performed before and after the study to measure serum cytokines, chemokines, IgE, quercetin and oxidized biomarkers. RESULTS: During the entire study period, total ocular score and ocular itching score for the EMIQ group were significantly lower (p < 0.05) than for the placebo group. When limited to the individual periods, total symptom score for the EMIQ group was significantly lower (p < 0.05, week 4-5) than that for the placebo group while other scores for the EMIQ group, such as total nasal score (p = 0.06, week 4-5), nasal obstruction score (p = 0.08, week 4-5), lacrimation score (p = 0.06, week 5-6), ocular congestion score (p = 0.08, week 4-7) and ADL score (p = 0.08, week 4-7), all tended to be lower. The levels of serum cytokines such as interleukin (IL)-4, IL-5, IL-12, IL-13, interferon-gamma, and eotaxin and IgE were not significantly downregulated by the intake of EMIQ but the serum concentrations of oxidized low-density lipoprotein and thymus and activation regulated chemokine were reduced. CONCLUSION: Intake of the quercetin glycoside EMIQ was safe and influenced ocular symptoms caused by pollinosis. PMID- 19295241 TI - Role of mammalian chitinases in asthma. AB - Asthma is a chronic inflammatory disease characterized by airway inflammation, mucus hypersecretion and airway hyperresponsiveness. Mechanisms underlying the pathogenesis of asthma are not fully understood. In recent years, there are mounting evidences demonstrating that mammalian chitinases may play a key role in mediating the T-helper 2 cell-driven inflammatory response that is commonly associated with asthma. Chitinases (e.g., chitotriosidase and acidic mammalian chitinase) are enzymes that degrade chitin, the second most abundant biopolymer that can be found in the cell walls of fungi, microfilarial sheaths of helminths, and exoskeletons of insects and crustaceans. There are also chitinase-like proteins (e.g., YKL-40, Ym1 and Ym2) that lack chitinolytic activity but retain chitin-binding ability. Therefore, chitinases were originally believed to function in host defense against parasitic infections, but the first discovery of their role in inflammatory airway diseases came as a surprise. There is ample evidence to support an association of acidic mammalian chitinase and YKL-40 with allergic bronchial asthma in patients. Our recent studies in a mouse asthma model revealed that anti-inflammatory drugs like corticosteroid and cysteinyl leukotriene receptor antagonist were able to suppress elevated pulmonary levels of mammalian chitinases. Taken together, mammalian chitinases may be useful as biomarkers for asthma. Notwithstanding, large-scale multi-center association studies are required to confirm this hypothesis. Besides, substantially more works using knockout mice, recombinant chitinases and siRNA technology are required to investigate a potential role of chitinases in the pathogenesis of asthma. PMID- 19295242 TI - Aspirin-induced COX-2 overexpression in monocytes of aspirin-intolerant patients. AB - BACKGROUND: We hypothesize that alternate regulation of cyclooxygenase-2 (COX-2) may predispose patients to aspirin-induced exacerbations.Therefore, we want to examine the dynamics of COX-2 up-regulation in whole blood monocytes in the presence and absence of aspirin. METHODS: COX-2 expression was evaluated by flow cytometry through intracellular staining of whole blood monocytes with anti-COX-2 antibodies. Enzyme up-regulation was monitored after in vitro stimulation with lipopolysaccharide (LPS) and/or aspirin in 19 aspirin-intolerant (AI) patients (8 aspirin-sensitive asthmatics and 11 urticaria-angioedema patients) and 14 healthy controls. RESULTS: We found significantly higher COX-2 expression levels after stimulation with LPS and aspirin (mean 78.8, range 44.9-92.3; p = 0.0002) in comparison to LPS alone (mean 65.9%, range 33.6-82.6) in AI patients. A comparable, but lower up-regulation was also observed after aspirin stimulation alone (median 2.1%, range 0.5-15.9; p = 0.004) in comparison with baseline values (median 1%, range 0.1-5.4). There was no significant difference in COX-2 expression between LPS and aspirin stimulation (mean 61.8%, range 26.8-89.2; p = 0.09) and LPS stimulation (mean 55.5%, range 28.1-74.3) nor between aspirin stimulation alone (median 0.5%, range 0-8.6; p = 0.8) and baseline values (median 0.4%, range 0-5.4) in healthy control subjects. CONCLUSIONS: The main finding of this study is that COX-2 appears to be differentially regulated in aspirin sensitive patients. What is really new is the observation that aspirin and LPS increase COX-2 expression on blood monocytes of AI asthmatics, a finding in contrast with the lack of an effect of the same stimuli on COX-2 expression on monocytes from healthy subjects. PMID- 19295244 TI - Does lymphatic involvement impact survival of early stage cervical cancer patients? PMID- 19295243 TI - Conduction failures in rabbit saphenous nerve unmyelinated fibers. AB - Recent experimental and theoretical data indicate that the functional capabilities of axons with specialized structures are much more diverse than traditionally thought. However, few observations were concerned with the main axons without arborization. In the present study, electrical stimulation of the saphenous nerve at different frequencies (2, 5, 10, 20 Hz) was used to test the role of activity-dependent effects on the pattern of action potentials that propagate along individual unmyelinated fibers (C fibers) within the trunk of the saphenous nerve in rabbits. Three basic types of C fiber responses to repetitive stimulation were observed: type-1 fibers showed an entrained response without conduction failure; type-2 fibers discharged with intermittent conduction failures; while only sporadic conduction failures happened in type 3. The failure modality in type-2 and type-3 fibers is closely related to the conductive distance as well as the frequency and duration of stimuli which lead to a critical level of conduction velocity slowing. A novel fluctuation in interspike intervals was always observed immediately before the occurrence of the failures, implying that the fluctuation of conduction velocity is correlated with imminent failures. Both the 4-aminopyridine-sensitive potassium current and hyperpolarization-activated cation current were recognized to be involved in the regulation of conduction failure patterns. The results confirmed, at least in part, the existence of conduction failures in the main axon of C fibers, suggesting that axonal operations may also be determinants for adaptation phenomenon and information processing in peripheral nervous system. PMID- 19295245 TI - Can dynamic MR imaging predict response in patients with rectal cancer undergoing cetuximab-based neoadjuvant chemoradiation? AB - BACKGROUND: The aim of this study was to evaluate therapy response in patients undergoing cetuximab-CapIri-based chemoradiation for rectal cancer using dynamic magnetic resonance imaging (dMRI). PATIENTS AND METHODS: The volumetric degree of tumor regression and contrast media perfusion were compared to the results of the histopathologic ypTN staging. 33 patients were examined using a 1.5-T scanner with repetitive 2D FLASH sequences after contrast media application. All patients were examined twice - before therapy and immediately before surgery. RESULTS: In all patients, the tumor volume decreased (mean 72 +/- 16%). In 25/33 patients, the slope of the contrast media enhancement curve decreased (mean 31 +/- 20%). In histopathologically proven downstaging after therapy, the decrease in slope was significantly higher than in the group without downstaging, and the decrease in slope was better for distinguishing between 'responder' and 'non-responder' than the decrease in volume. CONCLUSION: Using dMRI helps to identify responders undergoing cetuximab-based chemoradiation better than volume decrease alone. PMID- 19295246 TI - Scarless endoscopic papillomectomy of the breast. AB - BACKGROUND: Efforts have been made to improve minimally invasive breast surgery techniques, resulting in less tissue damage and much better cosmetic results. We evaluated the therapeutic value of a new scarless operation, endoscopic papillomectomy (EP), in patients with pathologic nipple discharge (PND). METHODS: Breast ductoscopy was performed on 126 women with PND. These patients underwent a variety of appropriate ductoscopy-assisted (DA) endosurgical interventions, combined with cytologic examinations. Success was determined by recurrence of PND and by standard radiological examinations. RESULTS: Ductoscopy was successfully performed in 102 patients. Of these 102 patients, 26 had solitary papillomas (SP), 5 had multiple papillomas (MP), 11 had intraductal debris, and 1 had a ductal epithelial surface abnormality with positive cytology. Of the 26 polypoid lesions (cytology negative), 22 were excised endoscopically (endoscopic papillomectomy). Patients with MP underwent DA-microdochectomy. Except in one patient, all discharges disappeared. After a mean +/- SD follow-up time of 11.5 +/- 5.8 months (range 2-22 months), there were no recurrences of nipple discharge and no radiological results suggestive of malignancy. Thus, the therapeutic efficacy of EP in our study was 95.4% (21/22). CONCLUSIONS: Ductoscopy is not only a diagnostic procedure, but is also therapeutic for breast papillomas. EP is a new scarless treatment option for patients with PND. PMID- 19295247 TI - Second-line therapy in refractory pancreatic cancer. results of a phase II study. AB - BACKGROUND: This phase II trial investigated the efficacy and safety of oxaliplatin (O), 5-fluorouracil (5-FU), and folinic acid (FA) (OFF) as second line treatment for patients with metastatic pancreatic adenocarcinoma after failure of first-line gemcitabine treatment. PATIENTS AND METHODS: 37 patients with confirmed progressive disease on gemcitabine therapy were treated with OFF (O 85 mg/m(2) days 8, 22; FA 500 mg/m(2), followed by 5-FU 2,600 mg/m(2) days 1, 8, 15, 22) every 6 weeks. Patients were treated on an outpatient basis and remained on treatment until disease progression. RESULTS: All patients were assessable for toxicity and effectiveness. We observed moderate hematotoxicity, the most common non-hematologic toxicity was neurotoxicity. A total of 12 patients had grade 3 nonhematologic toxicities: nausea and vomiting (4 patients), reversible neurotoxicity (5 patients), and diarrhea (3 patients). No grade 4 toxicities were observed. Median time to progression was 12 (1-125) weeks. Survival in second line was 22 (4-326+) weeks. Overall disease control rate was 49% (complete remission = 3%; partial remission = 3%; stable disease > 12 weeks = 43%). CONCLUSIONS: This regimen is feasible and active with an acceptable toxicity profile; it can be safely administered in an outpatient setting. There is an urgent need for further investigation in phase III trials. PMID- 19295248 TI - Impact of the ratio of metastatic to examined lymph nodes on the survival of early-stage cervical cancer patients. AB - BACKGROUND: The aim of the study was to determine the impact of the ratio of metastatic to examined lymph nodes (LN) on the survival of early-stage cervical cancer patients. METHODS: A retrospective review was performed of all patients undergoing primary radical hysterectomy and pelvicparaaortic lymphadenectomy for stage I-II cervical cancer at the Ankara Oncology Hospital from 1995 to 2000. Survival was determined by Kaplan-Meier method and differences were assessed by log-rank test. RESULTS: A total of 2,085 LN were examined. Nodal metastasis was found in 22.1% of the patients. Patients with zero involved LN had a 5-year survival of 89.19%, while patients with 1 positive LN and > 1 positive LNs had a 5-year survival of 100 and 76.92%, respectively. The difference in survival among these three groups was not statistically significant. The 5-year survival rates according to the metastatic/examined LN ratio (LNR) were 75, 90, and 89.19% in the groups of patients with LNR >10, 1-10, and 0%, respectively. The difference in survival among these three groups was not statistically significant. CONCLUSIONS: This factor might be helpful for a better prognostic discrimination of patients with metastatic LN. The stratification of node-positive cervical cancer for prognostic and treatment purposes warrants further investigation. PMID- 19295249 TI - Improved survival of patients with metastatic breast cancer in routine care: results of a retrospective study in a community-based oncology group practice 1995-2005. AB - BACKGROUND: Little is known about the treatment reality in metastatic breast cancer (MBC) outside clinical trials. We undertook this analysis to evaluate the actual treatment reality for unselected patients with MBC in routine care. PATIENTS AND METHODS: All patients with MBC, who were treated in our community based group practice between 1995 and 2005, were analyzed retrospectively concerning prognostic factors, treatment, and survival. RESULTS: 403 consecutive patients were evaluated with a median age of 60 years (range 32-93). Aromatase inhibitor therapy was used in 87% of all patients. 83% received chemotherapy with the median number of lines being 3 (1-15). An anthracycline was given to 49%, a taxane was used in 55%, vinorelbine in 42%, capecitabine in 36%, gemcitabine in 28%, and a platinum compound in 9%. 94% of patients with bone metastasis received a bisphosphonate, and 63% of HER-2/neu-positive patients were treated with trastuzumab. Median survival since the start of palliative therapy was 30 months. Statistical analysis revealed as major prognostic factors hormone receptor status and prevalence of only bone metastasis. CONCLUSIONS: Treatment reality of MBC in routine care reveals a prolonged median survival of 30 months which is probably due to the sequential use of the most effective treatment modalities. PMID- 19295250 TI - Fatal Clostridium septicum infection in a patient with non-Hodgkin's lymphoma undergoing multimodal oncologic therapy. AB - BACKGROUND: Clostridium septicum infections are rare but often fatal. They are known to be associated with a compromised immune system, e.g. with hematologic malignancies. CASE REPORT: We report on a spontaneous, non-traumatic C. septicum infection in a patient with non-Hodgkin's lymphoma undergoing irradiation due to abdominal bulk. Irradiation was delivered in daily fractions of 1.8 Gy. After 7 fractions, the patient suddenly developed severe pain in the lower leg. Clinical examination showed no pathological findings. However, a rapid progression of symptoms with motoric and sensoric deficits progressed rapidly within hours. Sizzling noise and gas entrainment were detectable about 9 h after the first symptoms. Despite immediate surgical treatment, and in the end amputation, the patient died 14 h later of septic shock. Histopathology showed gas gangrene in the leg tissue, with hemorrhagic muscular necrosis and enclosed gas vesicles. C. septicum was identified as causative organism. CONCLUSIONS: Irradiation as well as chemotherapy may have enhanced the risk by immunosuppression and induction of mucosal damage. Earlier antibiotic treatment might have slowed down the clinical course and thus given more time for correct diagnosis and treatment. PMID- 19295251 TI - Relapse of IgA lambda multiple myeloma presenting as obstructive jaundice and abdominal pain. AB - BACKGROUND: Only few cases of pancreatic involvement of multiple myeloma (MM) have been reported in the medical literature. PATIENTS AND METHODS: We here report a case of devastating extramedullary relapse of IgA/lambdaMM (stage IA) treated at diagnosis with a dexamethasone, adriamycin, vincristine (DAV) regimen followed by high-dose therapy and autologous stem cell transplantation (ASCT), achieving a partial remission. After 6 years of stable disease, the patient presented symptoms of obstructive jaundice determined by a large mass of the head of the pancreas. An ultrasound-guided fine-needle aspiration cytology of the pancreatic mass revealed the presence of myeloma plasma cells. A chest X-ray demonstrated a massive right pleural effusion, and the cytomorphologic evaluation of the pleural effusion showed the presence of abnormal plasma cells. RESULTS: We observed a progression of disease despite an aggressive treatment with high-dose cyclophosphamide. CONCLUSIONS: Our case shows that extramedullary relapses of MM after ASCT are very resistant to conventional chemotherapy. The role of new drugs and the optimal treatment strategy in these cases remain to be defined. PMID- 19295252 TI - A case of metachronous triple primary urogenital cancer: urinary bladder, prostate, and renal cancer. AB - BACKGROUND: Multiple primary malignant tumors are rarely seen. Tobacco is one of the factors in their etiology. We report the case of a heavy smoker with metachronous triple primary cancer occurring in the prostate, kidney and urinary bladder. CASE REPORT: A 70-year-old man with prostate cancer presented with the complaint of hematuria. Computed tomography (CT) showed increased wall thickness of the urinary bladder with an enlarged prostate. After the trans-urothelial resection operation pathological diagnosis was consistent with transitional cell carcinoma of the urinary bladder. After 9 months of follow-up, the control CT showed metastatic lesions in the right and left kidneys and in the right lung. Bilateral partial nephrectomy was performed. Interestingly, renal cell carcinoma (RCC) was diagnosed. Rightsided video-assisted thoracoscopic surgery was also performed. The results of the histopathological examination were consistent with metastatic RCC. CONCLUSIONS: Although the patient presented with triple carcinoma, there was no familial cancer history suggesting a genetic association. The patient was a heavy smoker, and tobacco usage may be the underlying cause of the detected cancers. This is one of the rare cases in the published literature with triple primary urogenital cancer. PMID- 19295253 TI - Cisplatin-based chemotherapy for pulmonary metastasized germ cell tumors of the testis--be aware of acute respiratory distress syndrome. AB - BACKGROUND: Cisplatin-based combination chemotherapy is regarded as standard of care for patients with advanced germ cell tumors. In patients with lung metastases and a high tumor load, an association between induction chemotherapy and the development of a 'tumorassociated' acute respiratory distress syndrome (ARDS) has been hypothesized. CASE REPORT: We report the clinical course of a 19 year-old patient who rapidly developed fatal ARDS during the first cycle of chemotherapy using the PEI regimen (cisplatin, etoposide and ifosfamide) for a metastasized (lung, liver, lymph nodes) germ cell tumor of the testis. CONCLUSION: Further clinical research in order to better define risk factors for developing ARDS in this patient population as well as novel strategies for the prevention and treatment of ARDS in those patients are necessary. PMID- 19295254 TI - Molecular targeted therapies for solid tumors: management of side effects. AB - This review will provide physicians and oncologists with an overview of side effects related to targeted agents that inhibit vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and mammalian target of rapamycin (mTOR) signaling in the treatment of solid tumors. Such targeted agents can be divided into monoclonal antibodies, tyrosine kinase inhibitors, multitargeted tyrosine kinase inhibitors and serine/threonine kinase inhibitors. Molecular targeted therapies are generally well tolerated, but inhibitory effects on the biological function of the targets in healthy tissue can result in specific treatment-related side effects, particularly with multitargeted agents. We offer some guidance on how to manage adverse events in cancer patients based on the range of options currently available. PMID- 19295255 TI - Segment-dependent activation of muscarinic acetylcholine receptor-mediated [35S]Guanosine-5'-O-(gamma-thiotriphosphate) binding in airway tissue membranes. AB - Muscarinic acetylcholine receptor (mAChR)-mediated guanine nucleotide-binding regulatory protein (G protein) activation and the functional interaction between receptors and the respective G proteins were investigated using an agonist induced [(35)S]guanosine-5'-O-(gamma-thiotriphosphate) ([(35)S]GTPgammaS)-binding approach in membranes of 3 native equine airway segments (trachea, bronchus and lung), which differ tremendously in mAChR density and subtype distribution; especially subtypes that couple negatively to adenylyl cyclase through G(i/0) proteins, i.e. M(2) receptors. The assay was initially optimized by determining the influence of incubation time, guanosine 5'-diphosphate (GDP), MgCl(2) and NaCl on basal and agonist-stimulated [(35)S]GTPgammaS binding. In standard assays, the presence of 10 mumol/l GDP, 10 mmol/l MgCl(2) and 200 mmol/l NaCl increased carbachol-induced specific [(35)S]GTPgammaS binding in a segment- and receptor-density-dependent manner. Hereby, mAChR agonists stimulated [(35)S]GTPgammaS binding with a rank order of potency (oxotremorine M > carbachol > acetylcholine), and in a specific segment- and receptor-density-dependent manner (trachea > bronchus > lung). The increase in the specific [(35)S]GTPgammaS binding was potently inhibited by the mAChR antagonist atropine. Pertussis toxin and N-ethylmaleimide as G(i/0) protein ADP-ribosylating and alkylating agents reduced basal and agonist-stimulated [(35)S]GTPgammaS binding. The mAChR stimulated [(35)S]GTPgammaS binding serves as a useful method for investigating the functional interaction between mAChRs and their respective G proteins in native airway tissue membranes of equines. PMID- 19295256 TI - Muscarinic receptor subtypes in the lower urinary tract. AB - Acetylcholine acting on muscarinic M(3) receptors on the detrusor muscle is the principal stimulus for inducing the contractile response for urinary bladder voiding. The urinary bladder expresses, however, all cloned muscarinic receptor subtypes (M(1)-M(5)). In terms of quantity, the M(2) subtype dominates over the M(3) subtype in the detrusor, and its role in contraction seems to be primarily indirect, by blocking stimuli from cAMP-coupled receptors that induce relaxation. The excitatory M(1) and inhibitory M(2) and/or M(4) subtypes are also expressed prejunctionally. Muscarinic M(1) and M(2)/M(4) autoreceptors facilitate and inhibit, respectively, the release of acetylcholine. The urothelium had been considered to be a passive barrier; however, during the last decade, it has been shown that the urothelium is of importance for bladder function. In a state of bladder pathology, muscarinic receptor changes occur in the detrusor, prejunctionally, and in the urothelium, but the character of the change differs between disorders. The urothelium expresses all subtypes of muscarinic receptors, and upon stimulation it releases factors affecting bladder afferents and smooth muscle. During inflammation, the expression of muscarinic M(5) receptors is increased, particularly in the urothelium, together with a cholinergic-induced production of nitric oxide in the mucosa. The present review describes signalling mechanisms, expression and functional effects of muscarinic receptors in the lower urinary tract. Their roles in physiological and pathophysiological conditions, as well as clinical implications of the occurrence of different muscarinic receptors, are discussed. PMID- 19295257 TI - [Recent progress in the field of hematopoietic stem cell transplantation for leukemia]. AB - The method and way of hematopoietic stem cell transplantation have been expanded and become complex recently. We discussed recent progress in this field, mainly in the treatment of leukemia. The first topic is the new arrival of the tyrosine kinase inhibitor, imatinib. Imatinib dramatically changed the treatment strategies for chronic myeloid leukemia. The same drug is now changing the treatment options and indications of hematopoietic stem cell transplantation for patients with Philadelphia chromosome-positive acute lymphoblastic leukemia. Efforts have also been made to expand the cell source for transplantation to cord blood and haploidentical donors. Almost 5,000 patients have undergone cord blood transplantation in Japan. Finally, the results of reduced-intensity stem cell transplantation may well expand the patient population eligible for transplantation. PMID- 19295258 TI - [Development of polymeric micelles for the delivery of antitumor agents]. AB - Recently, drug delivery systems(DDS)have received increasing attention as a medical application of nanotechnology. Among them, polymeric micelles, self assemblies of block copolymers, are a promising nanocarrier, because they can incorporate a variety of therapeutic agents such as hydrophobic substances, metal complexes and charged macromolecular compounds. Also, polymeric micelles have been demonstrated to effectively accumulate in the tumor tissue and show remarkable antitumor efficacy. Indeed, polymeric micelles encapsulating adriamycin, paclitaxel, cisplatin and SN-38 have progressed to clinical trials. This review deals with the development strategy and recent progress of polymeric micelles for the delivery of chemotherapeutic agents. PMID- 19295259 TI - [Peptide aptamers and nano-carriers for cancer therapy]. AB - Peptide aptamers are specific binders that are artificially created using in vitro evolution systems. The peptide aptamers that can detect malignant cells in vivo, or can bind surfaces of various materials will enable us to develop novel types of cancer diagnosis and cancer therapy agents by combining with various nano-carriers. Here, the development of peptide aptamer research is described in terms of artificial protein research. Also, two examples of nano-carriers are introduced, i. e., a ferritin, a natural caged protein, and carbon nanohorns, members of the carbon nanomaterial family, by focusing their potential roles in cancer diagnosis and cancer therapy. PMID- 19295260 TI - [Molecular imaging--diagnosis by functional imaging]. AB - In the 20th century, technological innovation in instrumentation has enabled us to noninvasively observe the structural detail of organs and tissues in diagnostic imaging. The 21st century is the era of analyzing the whole body at the molecular level as symbolized by post-genomic studies. In addition to structural information, functional information will be applied to the non invasive imaging of the human organ functions in the near future. Thus, the era of structural imaging will progress to the era of functional imaging. Recent progress in optical imaging has accelerated the multimodality of instruments for structural imaging such as magnetic resonance imaging(MRI)and computerized tomography( CT), and functional imaging with molecular probes. Although there are many limitations to optical modality for whole body imaging, the multi-functional nature of optics will cause a revolution in diagnostic imaging. PMID- 19295261 TI - [Cancer chemotherapy utilizing nanotechnology]. AB - Various attempts are under way to apply nanotechnology, in which remarkable progress has been made in recent years, to the diagnosis and treatment of cancer. For treating solid tumors, a system has been developed for more efficiently delivering nano-scale polymeric formulations to the tumors, utilizing the Enhanced Permeability and Retention( EPR)effect due to the characteristics of the tumor vascular system. Such nano-scale devices include a liposomal formulation, albumin-bound formulation and polymer micellar formulation, and each is currently under clinical development; some of these, such as doxorubicin-containing liposome(Doxyl), are already used in clinical practice. This paper describes the characterization of these nano-scale devices and ongoing clinical trials. PMID- 19295262 TI - [Clinical phase I trial of S-1 in the combination with DOC using super-selective intra-arterial infusion with oral cancer]. AB - The super-selective intra-arterial infusion, which has high anti-tumor effect to infuse high concentration of drugs into arterial in the control of tumor, has been expected to have local control. S-1, developed by the scientific theory of both potentiating antitumor activity of 5-fluorouracil(5-FU)and reducing gastrointestinal toxicity induced by 5-FU, is an active agent for squamous cell carcinoma of the head and neck(HNSCC). Docetaxel(DOC)is the drug Taxanes which has anti-tumor effect by mechanism different from conventional anti-tumor mechanism of action. In Yamaguchi University, DOC+CDDP+5-FU in the three-drug combination therapy shows high anti-tumor effect for advanced oral cancer. In the present study, we conducted a phase I study to examine local control of S-1 in the combination with DOC using super-selective intra-arterial infusion with oral cancer. The study performed super-selective intra-arterial infusion of DOC on the first day, and was considered as the schedule which prescribes three-week S-1 for patients every day from same day. Since blood toxicity nature grade 4 discovered the result in S-1: 65 mg/m(2)day and DOC: 50 mg/ m(2), we decided that recommended dose(RD)was S-1: 65 mg/m(2) and DOC: 40 mg/m(2). PMID- 19295263 TI - [Clinicopathological evaluation of preoperative chemotherapy with doxorubicin plus cyclophosphamide followed by paclitaxel for locally advanced breast cancer]. AB - While the optimum regimen and therapeutic duration of preoperative chemotherapy have not been established for advanced breast cancer, the combination regimen of anthracycline series and taxane series, which has been well received as a postoperative chemotherapy, is also recommended for preoperative chemotherapy. We here present a clinicopathological evaluation of the safety and efficacy of the regimen and the predictors of the effectiveness of preoperative chemotherapy conducted retrospectively in patients with advanced breast cancer who were treated preoperatively with the combination of doxorubicin and cyclophosphamide followed by sequential weekly paclitaxel during the period between January 2004 and June 2007 at Yamada Red Cross Hospital. Eleven patients were treated preoperatively with the regimen during the study period. Tumors shrank in all patients following the preoperative chemotherapy with a mean reduction rate of 64.1%. Adverse reactions to the preoperative chemotherapy included leukopenia; grade 3 in 1 patient and grade 2 or less in the remaining patients. Response rates evaluated in the primary tumors and lymph nodes were high. As for the predictors of the effectiveness of the regimen as preoperative chemotherapy, estrogen receptor(ER)and the presence of HER2 were investigated, and antitumor effects were high in patients with negative ER and positive HRE2. PMID- 19295264 TI - [Clinical significance of intratumoral TS levels and DPD activity in breast cancer]. AB - We evaluated the usefulness of the level of thymidylate synthase(TS)and dihydropyrimidine dehydrogenase(DPD) activity as prognostic factors and indicators for selection of chemotherapy regimens. Between November 1997 and March 1999, fifty-seven patients with stages I - IIIa primary breast cancer were registered. Using recurrence risk categories, they were classified into TAM monotherapy, TAM+oral 5-FU, and TAM+CMF groups(each were standard regimens at the time), and underwent postoperative adjuvant chemotherapy. The relationship between prognosis and the TS level and DPD activity, in addition to conventional risk factors, was examined. The recurrence-free survival time curve showed significant differences when stratified by tumor diameter, ER expression, and TS levels, but not by menopausal status, nodal status, surgical method, p53 expression, DPD activity, or HER2 expression. These results suggest that the TS level is useful as a prognostic factor for breast cancer. PMID- 19295265 TI - [The effectiveness of S-1 based sequential chemotherapy as second-line treatment for advanced/recurrent gastric cancer]. AB - BACKGROUND: We conducted this study to evaluate the efficacy of S-1 combination chemotherapy as second-line treatment for advanced/recurrent gastric cancer that was resistant to S-1 based chemotherapy as first-line treatment. PATIENTS AND METHODS: We evaluated patients included in phase II.III clinical trials, that is SPIRITS trial(S-1 vs CDDP +S-1), GC0301/TOP-002(S-1 vs CPT-11+S-1), OGSG0002(S 1+CPT-11)and OGSG0105(S-1+paclitaxel). Eligibility criteria at first-line included; pathologically proven gastric cancer, adequate bone marrow, hepatic, and renal functions, PS 0-2, no prior therapy, life expectancy estimated > or =12 weeks, age 20-75 years and written informed consent. Endpoints were as follows; )PFS in first-line and second-line, )Time to Second Progression(TSP), 3) OS. RESULTS: Sixty-six patients were evaluable in this study. We classified these patients into 4 groups according to the protocol. A)S-1 alone in first-line and S 1 combination in second-line(n=7), B)S-1 alone in first-line and other drugs in second-line(n=13), C)S-1 combination in first-line and another S-1 combination in second-line(n=33), D) S-1 combination in first-line and other drugs in second line(n=13). We compared S-1 combination group(A+C)to other drugs group(B+D)in second-line. In first-line, PFS was 157.5 days in group(A+C)and 130 days in group(B +D)(p=0.2749). In second-line, PFS, TSP and OS were as follows; 72.5, 256.5, 473 days in group(A+C)and 56, 201.5, 398.5 days(PFS; p=0.0806, TSP: p=0.0718, OS: p=0.0204)in group(B+D), respectively. With regards to adverse events, group(A+C)in first-line showed higher frequency in grade 3/4 leukopenia(10%), febrile neutropenia(5%)and grade 3 diarrhea(10%)than group(B+D). In second-line, group(B+D)showed grade 3/4 leukopenia (12%)and neutropenia(8%)than group(A+C). There were no treatment-related deaths. CONCLUSION: These results indicate that S-1 combination chemotherapy is efficient as second-line for advanced/recurrent gastric cancer that got resistant to S-1 based chemotherapy as first-line. PMID- 19295266 TI - [Expression of thymidine phosphorylase in primary colorectal cancer and liver metastasis-relationship between mRNA levels in cancer cells and protein levels in cancerous tissue]. AB - PURPOSE: To compare mRNA levels in cancer cells and protein levels in cancerous tissue in terms of the expression of thymidine phosphorylase(TP). MATERIALS AND METHODS: The materials were resected specimens of primary colorectal cancer(n=84)and synchronous liver metastasis(n=15). The expressions of TP mRNA in cancer cells was quantified by reverse-transcriptase polymerase chain reaction(RT PCR)using cells obtained by microdissection of paraffinembedded specimens(Danenberg tumor profiling method). The protein level of TP was determined by enzyme-linked immunosorbent assay(ELISA). RESULTS: There was a weak relationship between TP mRNA levels and TP protein levels in primary colorectal cancers(p=0.04), but these factors did not correlate with each other in liver metastases. There was no relationship between primary colorectal lesions and synchronous liver metastases in terms of TP mRNA levels and TP protein levels. In relation to the efficacy of 5-fluorouracil(or UFT)/Leucovorin, the mRNA level of the primary lesion was marginally higher in the order of patients with PD(n=11), SD(n=3), and CR/PR(n=4)(p=0.05). However, the TP protein level did not correlate with therapeutic efficacy. CONCLUSIONS: The different methods of determining the expression of TP in colorectal cancer specimens do not correlate with each other and may cause different therapeutic results after 5-fluorouracil-based chemotherapy. PMID- 19295267 TI - [Development of taste disorders following FOLFOX-FOLFIRI therapy and its effects on the QOL of patients with colorectal cancer]. AB - Taste disorders are frequent occurrences among those patients under the FOLFOX FOLFIRI regimen for colorectal cancer. We conducted a study on the development of taste disorders among colorectal cancer patients under this regimen and the effect of such disorders on their QOL. Taste disorders occurred in 58.1%(18/31 cases)of these patients and the disorders affected appetites in 50%(9 cases). The changes in taste sensations were subtle in most but some described certain tastes as exaggerated. Others reported changes in all taste sensations, including sweet, salty, bitter and sour, as well as deliciousness. When tested via the QOL Survey Sheet(Quality of Life Questionnaire for Cancer Patients Treated with Anticancer Drugs: QOL-ACD), the QOL was found to have deteriorated significantly in those who stated that taste disorders affected their appetite, in comparison with those who were unaffected. In patients with colorectal cancers and treated with the FOLFOX-FOLFIRI regimen, taste disorders are frequent occurrences. The poor nutritional state due to a loss of appetite may constitute a factor responsible for a lowering QOL. PMID- 19295268 TI - [Porous gelatin particles for hepatic arterial embolization; investigation of the size distribution and fragmentation before and after microcatheter passage]. AB - Liver abscess formation has been recently concerned after hepatic arterial embolization using porous gelatin particles( Gelpart)(GP), and inhomogeneous particles size or fragmentation of GP may partly contribute to this complication. The purpose of this study was to investigate the size distribution and incidence of fragments of GP before and after microcatheter passage. 1 mm- and 2 mm-GP were injected into three microcatheters with different inner-diameters by three interventional radiologists. The particles were stained with methylene blue dye, and the longer- and shorter-particle diameters were measured by a digital microscope. The particles less than 300 microm and 600 microm in longer- diameter was defined as fragment for 1 mm- and 2 mm-GP, respectively. Diameter of GP showed a wide bimodal distribution by fragments and non-fragment particles. The change of mean particle diameter ranged within 10%, and the ratio of shorter- to longer-particle diameter remained similarly at 0.75. The incidence of fragments was approximately 10% and 5% before microcatheter passage and increased by one two times and two-five times after microcatheter passage for 1 mm- and 2 mm-GP, respectively. In conclusion, GP were ununiform in particle size distribution and contained fragments. GP did not change in shape significantly after microcatheter passage, however, 2 mm-GP was more prone to fragmentation than 1 mm-GP. PMID- 19295269 TI - [Discussion of alleviating digestive medication toxicity in S-1 administered patients--retrospective and comparative study of the health records of continuously-administered patients and withdrawal patients]. AB - 79 patients(almost gastric cancer), who took drug S-1for the period from November 2000 to February 2006 in Asahi Rosai Hospital, were placed in two groups--a continuous group and a withdrawal group--to investigate and discuss the background factors contributing to the alleviation of digestive medication toxicity. The average number of days elapsed until drug withdrawal was 20 days, approximately 60% of the causes for withdrawal being attributable to digestive symptoms. A comparison between a subgroup of post-gastrectomy patients and a subgroup of non-gastrectomy patients showed that single S-1 drug administration could be continued for a longer period in the former than in the latter (p<0.05). A comparative study on drug regimens among all the patients demonstrated that a two-week regimen allowed the drug to be continuously administered for a longer period than a three- or four-week regimen(p<0.05). It was suggested that adverse effects might be alleviated by suppressing acid secretion in the stomach in the post-gastrectomy group. For the patients in that group, drug withdrawal is almost impossible. Considering that the average number of days elapsed until drug withdrawal is approximately 20 days, there seems to be a pressing need to establish the two-week regimen for continuous S-1 administration, which is not contained in the attachment hereto. PMID- 19295270 TI - [Efficacy of oral cavity care in preventing stomatitis (mucositis) in cancer chemotherapy]. AB - Stomatitis is a common side effect during cancer chemotherapy. We hypothesized that careful oral cavity care using patient guidance and cleanliness index prevents stomatitis in cancer chemotherapy. We introduced oral care patient guidance including teaching good brushing methods, O'Leary's Plaque Control Record(PCR)as a cleanliness index, and Eilers' Oral Assessment Guide(OAG)as an overall index after April 2006. We evaluated the incidence of stomatitis in 20 patients(10 patients between April 2004 to May 2006 and 10 patients after April 2006)with esophageal cancer who received chemotherapy including 5-FU and CDDP. Patients receiving brushing training after 2006 were evaluated regarding cleanliness of their oral cavities using PCR index and OAG index. The rates of stomatitis were 60%(6/10)and 40%(4/10)before and after the introduction of oral care patient guidance. The average of PCR index decreased from 82% to 46% after teaching good brushing method to the patients. The average of OAG index after brushing training was 9.14 which was better score compared with previous reports. Introduction of oral care patient guidance decreased the incidence of stomatitis. Both PCR and OAG indexes were useful in evaluating the objective condition of the oral cavity and in sharing patients' information among a medical team. These indexes encouraged the patients to clean their oral cavities. PMID- 19295271 TI - [Importance of the primary physician for pain management in patients with recurrent advanced cancer--a questionnaire survey]. AB - With the objective of clarifying points that needed improvement to provide earlier and better treatment of pain by assessing the current state of cancer pain management in Japan, we conducted a questionnaire survey about pain management in patients with advanced/recurrent cancer who were suffering from pain. The results of the survey revealed that it is important for primary physician to place greater emphasis on pain management when treating cancer patients, to inform patients that the doctor should always be told if the patient has pain, and provide appropriate information about medical narcotics to their patients. The team approach to management of cancer has been increasing in importance recently. This survey suggested it is important for primary physicians, who play a central role in such teams, to listen to their patients' complaints about symptoms including pain. Furthermore, it should be remembered that patients are eager to establish a good, trusting relationship with their primary physician. PMID- 19295272 TI - [A case of breast reconstruction surgery using flap of latissimus dorsi after neoadjuvant chemotherapy]. AB - We performed breast reconstruction surgery with mastectomy after neoadjuvant chemotherapy(NAC)for a patient with NAC indication desiring breast conservation. The case was a 34-year-old single woman. In March, 2007, she was aware of a lump in her left breast and visited our hospital. The diagnosis was solid-tubular carcinoma 3 cm in diameter from close examination. We performed preoperative chemotherapy with EC(epirubicin 90 mg/m(2), cyclophosphamide 600 mg/m(2))x4, followed by 3w-paclitaxel 175 mg/m(2)x4, and then performed mastectomy with axillary dissection and breast reconstruction surgery using the flap of latissimus dorsi at the same time. Pathologically, pCR was provided. We thought that there are many advantages to both treatment of breast cancer and the cosmetic characteristics. The patient was very satisfied. But further cumulative examinations are awaited because there is not much evidence at present. PMID- 19295273 TI - [A patient with axillary node metastasis from breast cancer who responded to trastuzumab/capecitabine combination therapy]. AB - We report an 83-year-old female with axillary node metastasis from breast cancer who responded to trastuzumab/ capecitabine combination therapy. In April 2007, she underwent surgery and at the same time axillary node metastasis was detected. As there was no hormone sensitivity, chemotherapy was selected, and administration of capecitabine at 1,800 mg/day(2 divided doses)and trastuzumab was initiated. Thoracic CT at the end of the six courses revealed the disappearance of the axillary node metastasis. No major side effects were produced. It was concluded from these findings that trastuzumab/capecitabine combination therapy could be safely and effectively administered to elderly advanced breast cancer patients. PMID- 19295274 TI - [Preoperative therapy using trastuzumab and weekly paclitaxel in a stage IIIB inoperable locally advanced HER2- positive breast cancer with complete pathologic response]. AB - A 63-year-old woman had a 12 cm tumor on her right breast with broad skin redness, satellite lesions and 8 cm ipsilateral lymph nodes swelling(T4bN2aM0, Stage IIIB). Core needle biopsy and immunohistochemistry of breast tumor showed invasive ductal carcinoma with negative hormone receptor(ER-, PgR-)and overexpression of HER2 (HercepTest 3+). She was treated with weekly paclitaxel(80 mg/m(2), 4 administrations with a week rest)and a com- bination with weekly trastuzumab(initially 4 mg/kg followed by 2 mg/kg every week, totally 11 administrations). After 3courses of administration, the breast tumor, skin redness and axillary swelling were completely disappeared(clinical complete response), then mastectomy with axillary dissection was performed. Histopathology of the breast and lymph nodes showed complete disappear of invasive cancer cells with only 2x1 mm residue of ductal component(pCR, grade 3, DC+). We conclude that the combination of weekly paclitaxel and trastuzumab is a promising neoadjuvant therapy regimen for HER2 positive, ER-negative breast cancer. PMID- 19295275 TI - [A case of sarcomatoid malignant peritoneal mesothelioma responding to combination chemotherapy of cyclophosphamide, vincristine, adriamycin and dacarbazine(CYVADIC)]. AB - A 66-year-old woman was seen at our hospital because of abdominal fullness. A computed tomography(CT)revealed massive tumors in abdominal cavity. The patient underwent surgery consisting of tumorectomy, segmental gastrectomy, partial resection of small intestin, transverse colectomy, left oophorectomy and gastrostomy. By using immunohistochemical staining, the patient was diagnosed as sarcomatoid malignant peritoneal mesothelioma. Rapidly abdominal fullness occurred as of 22 days after the operation, and an abdominal CT revealed the massive recurrent tumors. We started a combination chemotherapy of cyclophosphamide, vincristine, adriamycin and dacarbazine (CYVADIC). The recurrent tumors showed remarkable reduction after the two courses of CYVADIC chemotherapy. Although we next started carboplatin and paclitaxel combination chemotherapy, she died due to rapidly progression of the disease with disseminated intravascular coagulation after 132 days of the operation. Malignant mesothelioma, especially sarcomatoid mesothelioma, is known to have a poor prognosis. However, our case suggests that we could improve the prognosis of sarcomatoid malignant mesothelioma by aggressive chemotherapy. PMID- 19295276 TI - [Three (3) cases of advanced hepatocellular carcinoma (HCC) treated successfully by transcatheter arterial chemoembolization (TACE) using lipiodol and a fine- powder formulated cisplatin (DH)]. AB - Although great advances have been made in therapies for patients with hepatocellular carcinoma(HCC), the prognosis for advanced HCC remains poor because liver transplantation is not applicable. We report three(3)cases of HCC treated successfully by transcatheter arterial chemoembolization(TACE)using a suspension of Lipiodol and a fine powder formulation of cisplatin(DDPH). Case No. 1 was a 64-year-old man with multiple HCCs who had undergone several sessions of TACE using doxorubicin(ADM). During the course of the treatment, the HCC became intractable and residual tumors were observed repeatedly within a short period. He was then treated by TACE using DDPH instead of ADM. The tumor marker levels decreased in response to this treatment and no recurrence of HCC has been observed. Case No. 2 was a 71-year-old man who had been diagnosed with multiple HCCs in 2004. He was treated by TACE with ADM, but the procedure had to be repeated more than three(3)times due to residual tumors. Despite the treatment, the tumor grew gradually and a formation of tumor thrombus was observed in the inferior vena cava. Both the tumor and tumor thrombus reduced in size after TACE with DDPH. Case No. 3 was a 52-year-old man who had been monitored diabetes mellitus and chronic hepatitis at our hospital. Multiple HCCs were diagnosed in 2006. TACE with DDPH was performed as an initial therapy. The tumors shrank or disappeared in response to this treatment. These results propose that TACE with DDHP against advanced HCC is effective. PMID- 19295277 TI - [A case report--complete disappearance of multiple intrahepatic recurrences and portal thrombus in a case of hepatocellular carcinoma treated with UFT-E]. AB - Here, we present a case of recurrent hepatocellular carcinoma(HCC)with portal thrombus that disappeared within only two months by oral tegafur(UFT-E). A 82 year-old man had recurrent multiple HCCs, which caused liver dysfunction because of diminished portal flow by the portal thrombus. Hepatic arterial chemo embolization and continuous arterial CDDP-infusion chemotherapy did not show any efficacy. Next, oral UFT-E administration(low-dose, 300 mg/day)was begun, and after 1 month, PIVKA-II decreased and liver dysfunction was normalized. His complete response continued for 8 months until his death due to aspiration pneumonia and progressive dementia. Low-dose UFT-E administration may be considered a candidate in a case of HCC with no alternative treatment. PMID- 19295278 TI - [A case of bone marrow carcinosis from gastric cancer that presented hypocalcemia caused by zoledronic acid during the treatment of methotrexate/5-fluorouracil sequential therapy]. AB - The case was a 64-year-old man. He was diagnosed as gastric cancer, lymph node metastases, brain metastases, bone marrow carcinomas, and disseminated intravascular coagulation(DIC). He was started on methotrexate(MTX)/5- fluorouracil(5-FU)sequential therapy(weekly administration of MTX(100 mg/m(2), iv bolus)followed by 5-FU(600 mg/m(2), iv bolus)with a 3 h interval). DIC was resolved, and the tumor marker decreased remarkably. Four weeks later, he received zoledronic acid 4 mg to prevent skeletal complication. Next day, fatigue and anorexia onset. Six days later, laboratory data showed severe hypocalcemia. He was started on calcium gluconate 3.4 g/day. The calcium level was normalized in twelve days, and the symptoms were improved. MTX /5-FU therapy was resumed, and his condition remained stable. However, after the ninth dosage, he developed fatigue and low back pain, and the DIC relapsed. We started paclitaxel therapy. But it was not effective and he died ten days later. It was considered that careful attention to hypocalcemia is necessary when we use zoledronic acid for the bone marrow carcinomas treated with chemotherapy. PMID- 19295279 TI - [A case of an older adult patient with recurrent gastric cancer successfully treated by low-dose S-1 for a long time]. AB - An 88-year-old woman, who had undergone distal gastrectomy for Stage IIIA gastric cancer, was diagnosed with recurrent abdominal lymph node metastasis 11 months after surgery. Creatinine clearance of this patient calculated by Cockcroft-Gault method was 44 mL/min and renal function was impaired, so we reduced the administration dosage to 50 mg/day. S-1 was administered for two weeks followed by one week rest. This schedule induced grade 2 anorexia and fatigue after one week administration, so it was converted to one week administration followed by one week rest. By this dosage and administration schedule, a partial response(PR)was continued for about four years without any adverse reaction. This successful case might indicate that it was important for older adult patients with gastric cancer to consider not only appropriate reduction of the dosage but also shortening of S-1 administration. PMID- 19295280 TI - [A case of peritoneal serous papillary adenocarcinoma]. AB - A78-year-old female was admitted to our hospital with anorexia due to ascites. We were not able to diagnose by cytological diagnosis. For a definite diagnosis she underwent an open biopsy. Histological findings revealed observable papillary adenocarcinoma with psammoma body, carcinoma cells were positive for Ber-EP4 on immunostaining, and ovaries were normal. Thus, we made a diagnosis of peritoneal serous papillary adenocarcinoma. She was treated by intraperitoneal administration of CDDP(20 mg/day: day 1, every four weeks). Ascites vanished and CA125 was within the normal range. There was no recurrence after 3 years, but Virchow's lymph node metastasis occurred. The patient is still alive 12 months after metastasis with CDDP intraperitoneal administration. PMID- 19295281 TI - [A case of resection of synchronous multiple liver metastases from colorectal cancer after FOLFOX chemotherapy]. AB - A 41-year-old man with multiple liver metastases from sigmoid colon cancer received a radical resection of the original tumor and 16 courses of weekly high dose 5-FU(WHF)chemotherapy via hepatic arterial reservoir. The metastatic lesions showed stable disease(SD), and systemic chemotherapy by mFOLFOX6 was begun via CV port. After 14 courses of mFOLFOX4, abdominal CT revealed liver metastases were remarkably reduced in size. Hepatic resection of lateral segment and radio frequency ablation(RFA)for S6 were enforced, and the patient was uneventfully discharged. Pathological findings of lateral segment revealed no residual cancer cells, indicating that the histological effect of mFOLFOX6 was Grade 3. PMID- 19295282 TI - [A case of ascending colon cancer with local recurrence responding completely to alternating modified-FOLFOX6 and FOLFIRI regimens(modified-FIREFOX regimen)]. AB - A 58-year-old woman underwent right hemicolectomy with lymph node dissection(D2)for advanced ascending colon cancer which pathological examinations revealed to be moderately-differentiated adenocarcinoma. CEA and CA19-9 levels increased 6 months after the operation. She started adjuvant chemotherapy with oral administration of UFT-E(400 mg/day), but CEA and CA19-9 levels continued to elevate. However, a recurrent tumor was not detected by computed tomography(CT)and endoscopic examinations. A local recurrence in the right lateral abdominal wall was confirmed by PET-CT examination. We then conducted modified-FOLFOX6/FOLFIRI alternating regimen(modified- FIREFOX regimen). After this therapy, repeated PET-CT showed that the abnormal FDG-uptake concentration had disappeared, leading to a complete response(CR). The adverse event was grade 3 in leucopenia and grade 2 in gastrointestinal toxicity. She had maintained CR for the 12 months since undergoing chemotherapy. CEA and CA19-9 levels reduced to the normal range. We report this case with some review of the literature. PMID- 19295283 TI - [A fatal case of meningeal carcinomatosis in a stage IV rectal cancer patient who had long time survival by multi- line chemotherapy]. AB - A 65-year-old man was referred with Stage IV rectal cancer with lung and liver metastasis. He had done low anterior resection. One month after operation, he was administered anti-cancer drugs of FOLFOX4 protocol. Six months after operation, partial response was obtained of abdominal CT scan evaluation. Nine months after operation, FOLFOX4 was stopped by peripheral nerve injuries. FOLFIRI protocol started as second-line chemotherapy. Nineteen months after operation, he was diagnosed left hydro-nephrosis by retro peritoneal lymph-node metastasis. He was treated by urethral stenting, and FOLFOX protocol was re-started. Twenty one months after operation, FOLFOX4 was stopped by drug allergy, and changed to FOLFIRI protocol. Twenty five months after operation, He felt difficulty of walking and dizziness. He was diagnosed meningeal carcinomatosis by brain CT scan. He was administered glycerol and steroid. His neurological symptoms was improved. Although he died of disease progression at twenty six months after operation. PMID- 19295284 TI - [A case of PEP(BEP)-resistant ovarian dysgerminoma successfully treated by VeIP therapy]. AB - Ovarian germ cell tumors are malignant tumors which commonly develop during childhood, and which are sensitive to chemotherapy. We have had a case of germ cell tumors which showed resistance to first-line PEP(BEP)chemotherapy. As second line chemotherapy, VeIP therapy was used, because it is possible that this therapy is effective against recurrent testicular germ cell tumors. The patient was fourteen years old. She experienced acute abdominal pain and visited the hospital, where she was diagnosed with torsion of an ovarian tumor. An emergency laparotomy and right salpingoophorectomy were performed, the pathological diagnosis being stage Ia ovarian dysgerminoma G1. She was followed for two years until her serum hCG-CTP elevated to 1.4 mIU/mL. An MRI revealed an abnormal signal in the left ovary, so we diagnosed this as a recurrence of the dysgerminoma. Then she received chemotherapy PEP(BEP), but after eight months of PEP (BEP), her serum hCG-CTP was again elevated to 14.5 mIU/mL. A recurrence was detected with an MRI and PET-CT, and another laparotomy was performed. The recurrent region was detected in the left ovary. A left ovarian cystectomy was performed in which CDDP ip was used. After the operation, the patient again underwent chemotherapy. VeIP (vinblastine+ifosfamide+cisplatin)was chosen as the second-line regimen. After 6 courses of this therapy, she had a follow-up operation. No recurrence region was found in the pelvic area. She remains without recurrence of this disease 24 months after VeIP therapy. This case suggests that VeIP therapy might be an effective second-line therapy for patients with PEP(BEP) resistant ovarian dysgerminoma. PMID- 19295285 TI - [The introduction of the clinical study in the general hospital]. AB - Our hospital fights against a clinical study clearly. However, our hospital did not participate in a clinical study here. When I widened a clinical study in cooperation with the staff, I make a progress report. I started a clinical study only with a doctor first. Therefore it was recognized many problems happened and to promote a clinical study without cooperation of the work of many kinds. I studied with the staff to promote a clinical study and deepened understanding. Thereafter the clinical study was led by a nurse of the chemotherapy room; , in addition, was able to get various cooperation from a skiagrapher. When OGSG0603 began, the nurse and the pharmacist served to become the key than a doctor. I was able to promote a clinical study by cooperation of much staff. In addition, I was able to carry out the system improvement of each section by performing a clinical study. PMID- 19295286 TI - [Bevacizumab (Avastin)]. AB - Vascular endothelial growth factor(VEGF)is a key mediator in angiogenesis, an essential step in tumor growth and metastasis. Elevated levels of VEGF observed in solid tumors are correlated with worse clinical outcomes. Research has therefore focused on developing agents that target VEGF to inhibit tumor growth. Bevacizumab(Avastin), a recombinant humanized monoclonal antibody targeting VEGF. Bevacizumab with conventional chemotherapy has shown antitumor activity in several cancer types with acceptable toxicity. In April 2007, the Japanese Ministry of Health, Labour and Welfare approved bevacizumab for patients with unresectable colorectal cancer. PMID- 19295287 TI - Update on tracheobronchial anatomy and flexible fiberoptic bronchoscopy in thoracic anesthesia. AB - PURPOSE OF REVIEW: This review is focused on tracheobronchial anatomy and the use of flexible fiberoptic bronchoscopy in thoracic anesthesia. RECENT FINDINGS: A complete knowledge of tracheobronchial anatomy is a key factor in determining proper position of lung isolation devices, namely double-lumen endotracheal tubes and bronchial blockers. In addition, changes occur in tracheobronchial anatomy with age; therefore, it is very important that every anesthesiologist is familiar with these anatomical changes in order to recognize anatomical landmarks and perform a successful placement of lung isolation devices. Flexible fiberoptic bronchoscopy must be considered an art in the practice of thoracic anesthesia. SUMMARY: Recognition of tracheobronchial anatomy and familiarity with the use of flexible fiberoptic bronchoscope are key components while managing patients undergoing thoracic surgery and anesthesia. PMID- 19295288 TI - Advances in interventional pulmonology. AB - PURPOSE OF REVIEW: Interventional pulmonology is a rapidly expanding field offering less invasive therapeutic procedures for significant pulmonary problems. Many of the therapies may be new for the anesthesiologist. Although less invasive than surgery, some of these procedures will carry significant risks and complications. The team approach by anesthesiologist and pulmonologist is key to the success of these procedures. RECENT FINDINGS: Many modalities for central airway obstruction have emerged, including the expanding application of airway stenting procedures. Diagnostic bronchoscopy with ultrasound guidance promises great advances in lung cancer staging. New bronchoscopic treatments of asthma and emphysema are actively under investigation. Advances in anesthetic agents and techniques for interventional pulmonology procedures have also occurred. SUMMARY: This review is intended to familiarize the anesthesiologist with current and rising therapeutic modalities for pulmonary disease. Knowledge of interventional pulmonology facilitates planning and preparation for well tolerated and effective procedures. PMID- 19295289 TI - Update on selective lobar blockade during pulmonary resections. AB - PURPOSE OF REVIEW: This review is an update on the advantages that selective lobar blockade provides in thoracic surgical patients who had previous contralateral lobectomy or pneumonectomy. RECENT FINDINGS: Selective lobar blockade while allowing surgical exposure also improves oxygenation in patients who cannot tolerate complete lung collapse due to limited reserve or previous lung surgery. SUMMARY: Selective lobar blockade is an alternative technique to achieve lobar collapse in patients who have had a previous lobectomy or pneumonectomy and require thoracic surgery. This selective blockade can be accomplished by using a bronchial blocker or a double-lumen endotracheal tube. In patients with previous bronchial resection, recognition of tracheobronchial anatomy with the fiberoptic bronchoscope is mandatory to increase the successful placement and function of lung isolation devices. The advantages of a selective blockade are to avoid total lung collapse and only block the lobe(s) in which surgery takes place and to improve intraoperative oxygenation. PMID- 19295290 TI - Update on one-lung ventilation: the use of continuous positive airway pressure ventilation and positive end-expiratory pressure ventilation--clinical application. AB - PURPOSE OF REVIEW: The purpose of this review is to examine the evidence for and the clinical use of continuous positive airway pressure (CPAP) and positive end expiratory pressure (PEEP) for the management of one-lung ventilation during thoracic surgery. CPAP and PEEP use are important as we are increasingly challenged with patients with less respiratory reserve and greater comorbidity leading to the need for greater clinical management and more interventions during one-lung ventilation for thoracic surgery to prevent perioperative complications. RECENT FINDINGS: The focus of this article is on the most recent literature with selected classic articles. First, the supportive literature and rationale for application of PEEP, CPAP or both during thoracic surgery are reviewed, relative to the threats of hypoxemia, hyperoxia and mechanical lung injury. The second part of the article focuses on the clinical use of PEEP and CPAP. Algorithms for the application of CPAP and PEEP to patients both at risk and not at risk of acute lung injury are presented. SUMMARY: CPAP and PEEP are useful not only to treat hypoxia and atelectasis as the consequence of one-lung ventilation, perhaps more importantly, also as part of a protective lung-ventilation strategy to ameliorate mechanical stress and prevent acute lung injury. PMID- 19295291 TI - Right heart function in thoracic surgery: role of echocardiography. AB - PURPOSE OF REVIEW: As part of the preoperative evaluation, echocardiography provides noninvasive assessment of cardiovascular status in patients scheduled for lung resection, especially in the presence of chronic elevation of pulmonary arterial pressures. The goal of this review is to summarize the recent literature on the topic. RECENT FINDINGS: Changes in right ventricular function can occur acutely during lung transplantation or occasionally during lung resection. In the postoperative period, changes in right heart function will depend on preexisting pulmonary hypertension, and whether it is exacerbated by worsening chronic obstructive pulmonary disease, pneumonia or development of the adult respiratory distress syndrome. Currently, it remains controversial whether routine lung resection leads to clinically significant changes in right heart function. SUMMARY: The use of echocardiography in the perioperative setting can be useful in diagnosing and treating right ventricular dysfunction, especially when associated with hemodynamic instability unresponsive to conventional treatment, or arrhythmias, which all may occur after lung resection. PMID- 19295292 TI - Postoperative care after pulmonary resection: postanesthesia care unit versus intensive care unit. AB - PURPOSE OF REVIEW: In an effort to maximize resource utilization and contain costs, immediate postoperative care after noncardiac thoracic surgery is often done in either the postanesthesia care unit or dedicated step down units, leaving the ICU for complex surgical cases, overtly high-risk patients, or the treatment of severe postoperative complications. This review analyzes the current modalities affecting length of stay and costs, mainly by allocating patients after elective lung resection to different postoperative areas according to their needs. RECENT FINDINGS: Several surgical models have been published in recent years with the goal of optimizing perioperative patient care and subsequently decreasing hospital costs and length of stay. The main focus has been on elective lung resection for lung cancer. Preoperative evaluation, changes in surgical and anesthetic techniques as well as careful planning on where to recover these patients seem to make a clinical and financial impact. SUMMARY: The development of models to help predict elective ICU admission should facilitate optimal care, cutting costs and shortening length of stay after lung resection. PMID- 19295293 TI - Open or minimally invasive esophagectomy: are the outcomes different? AB - PURPOSE OF REVIEW: Since the beginning of the 1990s, the use of minimally invasive esophagectomy instead of the open technique has increased. Should this type of approach change the way we manage anesthesia for a patient undergoing esophagectomy for cancer? RECENT FINDINGS: Because valid direct comparisons with open surgery are lacking, one cannot make definitive statements regarding the potential benefits of minimally invasive surgery. Rough comparisons with recent reports on open surgery suggest that reduced mortality, respiratory complications and blood loss, plus a more rapid return to a good quality of life are areas in which minimally invasive surgery might prove superior. Leak rates were similar to those reported with open procedures. Surprisingly, length of hospital stay and overall morbidity are similar with both techniques. Reported operating times appear longer than one might expect for open operations, which mirrors the experience of laparoscopic procedures in other areas. SUMMARY: The implantation of minimally invasive esophagectomy seems inevitable in spite of the absence of randomized, controlled trials. The use of the prone position with one lung ventilation during minimally invasive esophagectomy seems positive. Protective ventilation during one lung ventilation may help to prevent pulmonary complications. Finally, the well accepted use of thoracic epidural anesthesia now has a new positive role following esophagectomy, improving the perfusion at the anastomotic level. PMID- 19295294 TI - Acute lung injury and outcomes after thoracic surgery. AB - PURPOSE OF REVIEW: The present review evaluates the evidence available in the literature tracking perioperative mortality and morbidity as well as the pathogenesis and management of acute lung injury (ALI) in patients undergoing thoracotomy. RECENT FINDINGS: Over the last decade, despite increasing age and comorbid conditions, the operative mortality has remained unchanged for patients undergoing lung resection, whereas procedure-related complications have declined. Better clinical outcomes are achieved in high-volume hospitals and when procedures are performed by a thoracic surgeon. Postthoracotomy ALI has become the leading cause of operative death, its incidence has remained stable (2-5%) and earlier diagnosis can be made by assessing the extravascular lung water volume with the single-indicator dilution technique. The pathogenesis of ALI implicates a multiple-hit sequence of various triggering factors (e.g. oxidative stress and surgical-induced inflammation) in addition to injurious ventilatory settings and genetic predisposition. SUMMARY: Knowledge of the perioperative risk factors of major complications and understanding of the mechanisms of postthoracotomy ALI enable anesthesiologists to implement 'protective' lung strategies including the use of low tidal volume (VT) with recruitment maneuvers, a goal-directed fluid approach and prophylactic treatment with inhaled beta2 adrenergic agonists. PMID- 19295295 TI - Cardiac output monitoring. AB - PURPOSE OF REVIEW: The primary goal of hemodynamic therapy is the prevention of inadequate tissue perfusion and inadequate oxygenation. Advanced cardiovascular monitoring is a prerequisite to optimize hemodynamic treatment in critically ill patients prone to cardiocirculatory failure. The most ideal cardiac output (CO) monitor should be reliable, continuous, noninvasive, operator-independent and cost-effective and should have a fast response time. Moreover, simultaneous measurement of cardiac preload enables the diagnosis of hypovolemia and hypervolemia. RECENT FINDINGS: Over recent years, a number of significant studies in the field of CO monitoring have been published. The available CO monitoring techniques can be divided into invasive techniques, minimally invasive techniques, and noninvasive techniques. SUMMARY: Minor invasive arterial thermodilution is the standard for the estimation of CO. Less invasive and continuous techniques such as pulse-contour CO and arterial waveform analysis are preferable. The accuracy of noncalibrated pulse-contour analysis is still a matter of discussion, although recent studies demonstrate acceptable accuracy compared with a standard technique. Doppler techniques are minimally invasive and offer a reasonable trend monitoring of CO. Noninvasive continuous techniques such as bioimpedance and bioreactance require further investigation. PMID- 19295296 TI - High thoracic epidural anaesthesia for cardiac surgery. AB - PURPOSE OF REVIEW: Epidurals have been used for cardiac surgery for more than 20 years. The worldwide-published use is now large enough to determine that there is no additional risk for epidural use in cardiac versus noncardiac surgery. RECENT FINDINGS: Several large case series have been added to the literature without cases of spinal damage. The estimated risk of epidural haematoma is 1: 12 000 (95% confidence interval of 1: 2100 to 1: 68000), which is comparable to noncardiac surgery. The fear of an increased risk of epidural haematoma associated with cardiopulmonary bypass has not eventuated. Improved analgesia, reduced pulmonary complications and reduced atrial fibrillation in off-pump coronary surgery have been reported. There are some case series and numerous case reports of awake cardiac surgery performed under epidural anaesthesia. This review will focus on safety, benefits and the logistics of performing epidural anaesthesia for cardiac surgery. SUMMARY: Fear of an increased risk of epidural haematoma has largely prevented increased use of this technique for cardiac surgery. Clinicians can be reassured that the risk of epidural use in cardiac surgery is similar to that for noncardiac surgery, which provides a new platform for considering risk versus benefit in their practice. PMID- 19295297 TI - Anesthesia in adult patients with congenital heart disease. AB - PURPOSE OF REVIEW: Recent advances in prenatal diagnosis, interventional cardiology, pediatric cardiac surgery, anesthesia, and critical care have resulted in an increasing number of adult patients with congenital heart disease (CHD). Most of these patients will require noncardiac surgery thus presenting a new challenge for anesthesiologists. The purpose of this article is to summarize preoperative and intraoperative implications for the anesthesiologist in the noncardiac surgery setting. RECENT FINDINGS: These patients present with an increased risk of perioperative mortality. One of the most specific recommendations from the American College of Cardiology conference published in 2001 was that adult patients with moderate-to-severe CHD undergoing noncardiac surgery should be referred to an adult CHD center with the consultation of an expert anesthesiologist. However, though most experts agree that grown-up CHD poses an increasing risk for noncardiac surgery, no major study focusing on this topic has yet been performed. SUMMARY: The number of adult patients with CHD is now superior to the number of children. This is a new challenge for anesthesiologist in the noncardiac surgery settings. PMID- 19295298 TI - Efficacy and safety of activated recombinant factor VII in cardiac surgical patients. AB - PURPOSE OF REVIEW: Excessive bleeding is a common and morbid problem after cardiac surgery. There is no doubt a need for an effective and safe hemostatic agent in order to minimize transfusions and avoid surgical reintervention for hemostasis. Recombinant activated factor VII (rFVIIa) is being used (off-label) increasingly after cardiac surgery to prevent or to control hemorrhage, but its efficacy and safety remain unclear. RECENT FINDINGS: Several case reports, case series and registries would tend to support the use of activated recombinant factor VII to control excessive bleeding after cardiac operations. On the contrary, two randomized controlled trials have produced negative results whereas a third has not been published yet. Adverse thrombotic events are reported with increasing frequency. SUMMARY: At present, the generalized use of rFVIIa to prevent or to control excessive bleeding after cardiac surgery cannot be recommended. The decision to administer a potent hemostatic such as rFVIIa outside its recognized prescribing indications should be made with caution by well informed physicians and discussed with the patient. Patients should be informed about knowledge gaps and pertinent risks, which are both important in the case of rFVIIa. PMID- 19295299 TI - Current aspects of perioperative fluid handling in vascular surgery. AB - PURPOSE OF REVIEW: Perioperative fluid management influences patient outcome. Vascular surgery unites various surgical procedures, mainly with a high impact on patients who often have relevant preexisting illnesses. There are only scarce data on this specialty, forcing the clinician to extrapolate existing data when planning perioperative fluid management. This review aims to summarize the underlying facts. RECENT FINDINGS: Perioperative insensible perspiration does not exceed 1 ml/kg per hour. A third space shift does not exist; therefore, its primary substitution is erroneous. Rather, a crystalloid fluid excess causes a tremendous shift towards the interstitial space. Colloidal volume effects are context sensitive, that is, only their use as a substitute when blood or plasma loss occurs leads to maintenance within the circulatory space. Colloidal hypervolaemia and surgical trauma both have the potential to deteriorate the vascular barrier, leading to plasma loss into the interstitial space. Current perioperative fluid therapy should aim to maintain normovolaemia of the individual body fluid compartments as far as possible. This might be achieved by combining a protocol-based replacement of extracellular losses (urinary output plus insensible perspiration) with isotonic balanced crystalloids and blood volume optimization using isooncotic colloids. SUMMARY: The basis of fluid therapy in vascular surgery is a careful differential indication of the respective classes of preparations. A goal-directed approach might help to avoid hypovolaemia. PMID- 19295300 TI - Anaesthesia for noncardiac surgery in the heart transplant recipient. AB - PURPOSE OF REVIEW: Anaesthetizing a patient who benefited from a heart transplant will undoubtedly fascinate any anaesthetist. Basic principles need to be adhered to in order to ensure that no unnecessary harm is caused to these patients. Furthermore, the conscientious anaesthetist will want to remain abreast of the advances in the understanding of physiological changes occurring over the years after transplantation and the continuous innovations in the field of immunosuppression. RECENT FINDINGS: Understanding of the mechanisms surrounding cardiac allograft vasculopathy and insight into the possibility of reinnervation are continuously evolving; but changes in therapeutic regimens in an attempt to lower side effects have probably a greater impact on day-to-day clinical practice. CONCLUSION: Understanding how to administer fluids adequately to a transplant patient; how to use direct vasoactive agents; how to manage the infectious risk; and the need to be aware of the multiple side effects of immunosuppressive regimens are essential to recognize, prevent or treat possible perioperative complications. PMID- 19295301 TI - Ca2+: is there something new for the cardiovascular anesthesiologist? AB - PURPOSE OF REVIEW: Anesthesiologists are frequently called upon to treat abnormalities of heart rhythm or pumping ability. Intracellular Ca is crucial for normal excitation-contraction coupling in the heart and plays a major role in the sequence of events that starts with an electrical signal generated in the atria and ends with myocardial contraction. RECENT FINDINGS: From controlled diffusion within the cell to a potential role as a biological clock, intracellular Ca is receiving a great deal of attention. For example, the pacemaking electrical signal is known to originate in the sinoatrial node myocyte, but exactly what role Ca plays is controversial despite the fact that the sinoatrial node was discovered over 100 years ago. Basic mechanisms involved in disease processes such as atrial fibrillation and new interventions for heart rate control are beginning to emerge. New discoveries in ventricular myocytes are also stimulating the development of promising therapeutic interventions to safely increase the pumping ability of the heart. SUMMARY: As our understanding of cardiac physiology and pharmacology progresses at the subcellular and molecular levels, new therapies will continue to emerge and the practice of anesthesia will benefit greatly. PMID- 19295302 TI - Aprotinin: 1 year on. AB - PURPOSE OF REVIEW: The nonspecific protease inhibitor aprotinin has been used successfully to reduce bleeding in cardiac surgery. Recent investigations have questioned its safety, and aprotinin has finally been withdrawn from marketing after a large prospective study demonstrated a trend toward higher mortality. RECENT FINDINGS: The initial studies of Karkouti and Mangano provoked a considerable number of large-scale investigations focusing on the safety issues of aprotinin. These studies were of retrospective nature and used sophisticated statistical methods to overcome a possible selection bias. Recently, aprotinin was predominantly used in patients with a higher risk of bleeding, which hampers a retrospective comparison with patients without the drug. This review summarizes the diverging results of these studies. SUMMARY: It remains a matter of speculation whether the quality and results of published data justify the withdrawal of aprotinin; however, one has to accept that this drug is no longer available. It is clear from the aprotinin story that there are no effective instruments to control the safety and clinical efficacy of a drug after its regulatory approval. This highlights the urgent need for independent clinical safety studies after the formal registration of a drug. PMID- 19295303 TI - The decade of polyomavirus BK-associated nephropathy: state of affairs. AB - In the last 10 years, better immunosuppression drugs have decreased the rates of acute rejection in kidney transplantation but have also led to the emergence of polyomavirus-associated nephropathy (PVAN). This occurs in 1% to 10% of patients with kidney transplantion and is caused by BK virus in more than 95% of cases. Less than 5% of cases are attributed to the JC virus. Initially, lack of recognition or late diagnosis of PVAN resulted in rapid loss of graft function in more than 50% of patients. In recent years, it has become clear that early diagnosis and timely reduction in immunosuppression is the only proven measure, which significantly affects the outcome of PVAN. Diverse interventions have been explored including the adjunctive use of cidofovir, leflunomide, fluoroquinolones, and intravenous immunoglobulins. Allograft histology is needed to definitively establish the diagnosis of PVAN, but is of limited sensitivity in the early stage of disease. Well-established techniques and protocols for systematic screening by urine cytology and quantitative molecular-genetic techniques allow now for timely intervention before irreversible parenchymal changes occur. Moreover, preemptive reduction in immunosuppression is most effective in presumptive PVAN as defined by surrogate markers (i.e., high BK virus viremia). In this setting, preservation of graft function can be considered the rule. Nevertheless, the recovery of BK virus-specific T-cell immunity may require prolonged periods during which cytopathic damage may continue to accumulate. Despite remarkable progress in the field, important challenges remain, such as the rare patient with PVAN refractory to any intervention and the newly recognized association of PVAN with urogenital tumors. PMID- 19295304 TI - Normothermic perfusion: a mini-review. AB - The quality of organ preservation is critical to the outcome of transplantation: preservation technology is a limiting factor in the further development in the use of marginal donor organs. Normothermic preservation (preservation at normal physiological temperature) enables prolonged preservation, resuscitation after warm ischemia and organ viability assessment before transplantation. PMID- 19295305 TI - Antiplatelet therapy in transplantation: new tricks from old drugs? PMID- 19295306 TI - One liver for four children: first clinical series of liver cell transplantation for severe neonatal urea cycle defects. AB - BACKGROUND: Urea cycle disorders (UCD) have a poor prognosis despite dietary and pharmacologic therapy, especially if the onset of the disease is within the neonatal period. They are promising target diseases for liver cell transplantation (LCT), which may be a less invasive alternative or supplementation to orthotopic liver transplantation. METHODS: Cryopreserved hepatocytes were isolated under good manufacturing practice conditions. Four children with severe neonatal UCD (age 1 day-3 years) received multiple intraportal infusions of cryopreserved hepatocytes from that same donor, a 9-day old neonate. Portal vein access was achieved surgically in two children, whereas the umbilical vein was suitable for interventional catheter placement in two neonates. Cell applications were carefully monitored by means of Doppler ultrasound and portal vein pressure. RESULTS: LCT was feasible in all children. No signs of portal vein thrombosis or extrahepatic shunting were observed. All children showed metabolic stabilization during observation periods of 4 to 13 months. One child with prenatally diagnosed ornithine transcarbamylase deficiency died after 4 months from a fatal metabolic decompensation. CONCLUSIONS: Given the poor prognosis of UCD with conservative therapy, LCT caused considerable beneficial effects. Periods of hyperammonemia and clinically relevant crises could be reduced during an observation period of up to 13 months. Though cell therapy is not a permanent therapeutic option, bridging to liver transplantation may be substantially improved. PMID- 19295307 TI - Comparison of transplantation of adipose tissue- and bone marrow-derived mesenchymal stem cells in the infarcted heart. AB - BACKGROUND: Mesenchymal stem cells hold promise for cardiovascular regenerative therapy. Derivation of these cells from the adipose tissue might be easier compared with bone marrow. However, the in vivo fate and function of adipose stromal cells (ASC) in the infarcted heart has never been compared directly to bone marrow-derived mesenchymal cells (MSC). METHODS: ASC and MSC were isolated from transgenic FVB mice with a beta-actin promoter driving firefly luciferase and green fluorescent protein double fusion reporter gene, and they were characterized using flow cytometry, microscopy, bioluminescence imaging and luminometry. FVB mice (n=8 per group) underwent myocardial infarction followed by intramyocardial injection of 5x10(5) ASC, MSC, fibroblasts (Fibro, positive control), or saline (negative control). Cell survival was measured using bioluminescence imaging for 6 weeks and cardiac function was monitored by echocardiography and pressure-volume analysis. Ventricular morphology was assessed using histology. RESULTS: ASC and MSC were CD34(-), CD45(-), c-Kit(-), CD90(+), Sca-1(+), shared similar morphology and had a population doubling time of approximately 2 days. Cells expressed Fluc reporter genes in a number dependent fashion as confirmed by luminometry. After cardiac transplantation, both cell types showed drastic donor cell death within 4 to 5 weeks. Furthermore, transplantation of either cell type was not capable of preserving ventricular function and dimensions, as confirmed by pressure-volume-loops and histology. CONCLUSION: This is the first study comparing the in vivo behavior of both cell types in the infarcted heart. ASC and MSC do not tolerate well in the cardiac environment, resulting in acute donor cell death and a subsequent loss of cardiac function similar to control groups. PMID- 19295308 TI - A novel JAK3 inhibitor, R348, attenuates chronic airway allograft rejection. AB - BACKGROUND: This study aimed at investigating the role of a novel JAK3 inhibitor, R348, in the prevention of chronic airway allograft rejection. METHODS: The heterotopic rat trachea transplant model was used. Recipients were treated daily with R348 (10, 20, 40, 80 mg/kg) or rapamycin (0.75 or 3 mg/kg). Blood levels of R348 and of its active metabolite R333 were measured. Grafts were harvested after 28 days to analyze epithelial morphology, mononuclear infiltration, and luminal obliteration. Plasma levels of circulating donor strain-reactive IgG antibodies were quantified. RESULTS: R348 was well tolerated at up to 40 mg/kg, but was toxic at 80 mg/kg. Blood levels of R333 at 2 and 24 hr were consistently 10 to 15 times higher than those of R348. Airway luminal obliteration after 28 days was significantly inhibited by R348 at 40 mg/kg (20.6%+/-13.2%, P<0.05) and 80 mg/kg (15.7%+/-7.6%, P<0.05) and by rapamycin at 3 mg/kg (11.6%+/-6.7% P<0.001) versus untreated controls (100%). R348 is more than or equal to 40 mg/kg but neither dose of rapamycin preserved the physiologic epithelial coverage with its prominent goblet cells population (8.8+/-1.5 goblet cells/microm circumference in syngeneic grafts and 8.0+/-0.9 and 4.3+/-1.2 with R348 80 mg/kg and 40 mg/kg, respectively). Peritracheal graft mononuclear infiltration was most effectively suppressed by R348 is more than or equal to 40 mg/kg (P<0.05) and rapamycin 3 mg/kg (P<0.01). Donor strain-reactive IgG antibodies were significantly decreased by R348 is more than or equal to 40 mg/kg (P0.05). Even subjects with low eGFR and microalbuminuria at baseline (n=10) maintained stable values posttransplantation (61.13+/-3.25 mL/min/1.73 m2 vs. 63.32+/-4.36 mL/min/1.73 m2, P=0.500). CONCLUSIONS: Kidney function remained stable after islet transplantation alone. The unchanged kidney function found in this sample may be attributed to healthier kidney status at baseline and possibly to prompt treatment of modifiable risk factors. Aggressive treatment of risk factors for nephropathy, such as blood pressure, low-density lipoprotein cholesterol, and careful tacrolimus levels monitorization, should be part of islet transplant recipient care. PMID- 19295313 TI - Early metabolic markers of islet allograft dysfunction. AB - BACKGROUND: Islet transplantation can restore normoglycemia to patients with unstable type 1 diabetes mellitus, but long-term insulin independence is usually not sustained. Identification of predictor(s) of islet allograft dysfunction (IGD) might allow for early intervention(s) to preserve functional islet mass. METHODS: Fourteen islet transplantation recipients with long-term history of type 1 diabetes mellitus underwent metabolic testing by mixed meal tolerance test, intravenous glucose tolerance test, and arginine stimulation test every 3 months postislet transplant completion. Metabolic responses were compared between subjects who maintained insulin independence at 18 months (group 1; n=5) and those who restarted insulin within 18 months (group 2; n=9). Data were analyzed before development of islet graft dysfunction and while insulin independent. RESULTS: The 90-min glucose, time-to-peak C-peptide, and area under the curve for glucose were consistently higher in group 2 and increased as a function of time. At 12 months, acute insulin release to glucose in group 2 was markedly reduced as compared with baseline (5.62+/-1.21 microIU/mL, n=4 vs. 16.14+/-3.69 microIU/mL, n=8), whereas it remained stable in group 1 (22.36+/-4.98 microIU/mL, n=5 vs. 27.70+/-2.83 microIU/mL, n=5). Acute insulin release to glucose, acute C-peptide release to glucose (ACpRg), and mixed meal stimulation index were significantly decreased and time-to-peak C-peptide, 90-min glucose, and area under the curve for glucose were significantly increased when measured at time points preceding intervals where IGD occurred compared with intervals where there was no IGD. CONCLUSIONS: The intravenous glucose tolerance test and mixed meal tolerance test may be useful in the prediction of IGD and should be essential components of the metabolic testing of islet transplant recipients. PMID- 19295314 TI - The changes of the medial right lobe, transplanted with left lobe liver graft from living donors. AB - BACKGROUND: Procurement of left lobe (LL) living donor graft with medial right lobe (mRL) might be an innovative technique. METHODS: The grafts were procured from a living donor, exposing the right anterior Glissonean pedicles, after confirmation of the demarcation line by test-clamping of the right Glissonean pedicle. Based on serial computed tomography, the increase in the graft volume (GV) after addition of mRL and changes in volumes were evaluated. RESULTS: The addition of mRL (n=7) increased GV by 48+/-9 g, which corresponded to a 4% increase in GV-to-standard liver volume ratio. After transplantation, mRL volume has increased in all cases. The regeneration rate of the mRL and other LL segments 1 month after transplantation was 61%+/-18% and 146%+/-15%, respectively. Viable hepatic parenchyma with marginal bile duct dilatations in transplanted mRL was observed in all the cases. Marginal enhancement was observed in those cases with promoted regeneration of transplanted mRL. In the cranial part of the mRL, portal branching from the left portal vein, over the middle hepatic vein, was observed in all cases. CONCLUSION: This technique affords an increase in GV in living donor LL procurement, and should increases the application of LL grafts in living donor liver transplantation. PMID- 19295316 TI - Quantitative patterns of azathioprine adherence after renal transplantation. AB - BACKGROUND: Renal transplant recipients regularly fail to take their prescribed immunosuppressive medications, frequently leading to adverse outcomes. METHODS: Medication vials incorporating electronic monitor circuits in their caps compiled prospective data files on the azathioprine dosing patterns of 180 adult renal transplant recipients monitored up to 4 years. These patients were followed for a mean of 8.7 years posttransplantation. RESULTS: Patients were divided into three groups by the medication doses missed during the first 6 months posttransplant. These initial dosing patterns remained remarkably consistent up to 4 years. Patients (n=47) missing the most doses (>or=5%) experienced earlier and more frequent acute rejection episodes (P=0.025). This group also demonstrated significantly longer interdose intervals (P=0.005), with more frequent (P<0.001) and longer (P<0.001) "drug holidays." A patient subgroup with early declining medication adherence (n=23) experienced dramatically poorer outcomes, with significantly increased acute rejection (P<0.001), chronic rejection (P=0.034), graft loss before death (P<0.001), and death (P=0.04). In all tertiles there was a trend toward missing more medication over time. CONCLUSIONS: Excellent posttransplant medication adherence is critical to improved outcomes. Individual dosing patterns are established early after hospital discharge and remain remarkably consistent, despite gradual erosion in adherence over time. The later consequences of medication nonadherence, especially early declines in adherence, include increased frequencies of rejection, graft loss, and death. PMID- 19295315 TI - IL10 and IL10 receptor gene variation and outcomes after unrelated and related hematopoietic cell transplantation. AB - BACKGROUND: Results of a previous study with human leukocyte antigen (HLA) identical siblings showed individual and synergistic associations of single nucleotide polymorphisms in the promoter region of the recipient's IL10 gene and the donor's IL10 receptor beta (IL-10RB) gene with development of grades III-IV acute graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation. METHODS: In this study of 936 patients who had unrelated donors, genotypes of single nucleotide polymorphisms in the IL10 gene and the IL-10RB gene were evaluated as correlates with outcomes after transplantation. RESULTS: We found no statistically significant associations of polymorphisms at positions 3575, -2763, -1082, and -592 of the IL10 gene or codon 238 of the IL10RB gene with severe acute GVHD, extensive chronic GVHD or nonrelapse mortality after hematopoietic cell transplantation. Among HLA-matched unrelated pairs, the patient's IL10/-592 genotype and donor's IL10RB/c238 genotype showed trends suggesting individual and combined associations with grades III-IV acute GVHD similar to those observed among patients with HLA-identical sibling donors. CONCLUSIONS: Although genetic variation in IL10 pathway affects risk of acute GVHD and non-relapse mortality in HLA-identical sibling transplants, the current results indicate that genetic variation in the IL10 pathway does not significant affect these outcomes in unrelated donor transplants suggesting that the strength of the alloimmune response in the latter exceeds the anti-inflammatory activity of IL10. PMID- 19295317 TI - Lack of association between beta-herpesvirus infection and bronchiolitis obliterans syndrome in lung transplant recipients in the era of antiviral prophylaxis. AB - BACKGROUND: Cytomegalovirus (CMV), human herpesvirus-6 and -7 (HHV-6 and -7) are beta-herpesviruses that commonly reactivate and have been proposed to trigger acute rejection and chronic allograft injury. We assessed the contribution of these viruses in the development of bronchiolitis obliterans syndrome (BOS) after lung transplantation. METHODS: Quantitative real-time polymerase chain reaction of bronchoalveolar lavage samples were performed for CMV, HHV-6 and -7 in a prospective cohort of lung transplant recipients. A time-dependent Cox regression analysis was used to correlate the risk of BOS and acute rejection in patients with and without beta-herpesviruses infection. RESULTS: Ninety-three patients were included in the study over a period of 3 years. A total of 581 samples from bronchoalveolar lavage were obtained. Sixty-one patients (65.6%) had at least one positive result for one of the beta-herpesviruses: 48 patients (51.6%) for CMV and 19 patients (20.4%) for both HHV-6 and -7. Median peak viral load was 3419 copies/mL for CMV, 258 copies/mL for HHV-6, and 665 copies/mL for HHV-7. Acute rejection (>or=grade 2) occurred in 46.2% and BOS (>or=stage 1) in 19.4% of the patients. In the Cox regression model the relative risk of acute rejection or BOS was not increased in patients with any beta-herpesviruses reactivation. Acute rejection was the only independently associated risk factor for BOS. CONCLUSIONS: In lung transplant recipients receiving prolonged antiviral prophylaxis, reactivation of beta-herpesviruses within the allograft was common. However, despite high viral loads in many patients, virus replication was not associated with the development of rejection or BOS. PMID- 19295318 TI - Mycophenolate and sirolimus as calcineurin inhibitor-free immunosuppression improves renal function better than calcineurin inhibitor-reduction in late cardiac transplant recipients with chronic renal failure. AB - BACKGROUND: Calcineurin-inhibitor-(CNI)-induced renal failure is one major cause of morbidity in cardiac transplantation (HTx). In this prospective, randomized, multicenter trial, the impact of immunosuppressive conversion toward CNI-free (mycophenolate mofetil [MMF] and sirolimus) or a CNI-reduced immunosuppressive regimen on renal function, efficacy, and safety was evaluated. METHODS: Since 2004, 63 HTx-patients (0.5-18.4 years after HTx) with CNI-based immunosuppression and reduced creatinine clearance less than 60 mL/min (39+/-15 mL/min) were included in this trial. Patients in the CNI-free-Group (group 1) were converted to sirolimus that was started with 2 mg/day until target trough levels (8-14 ng/mL) were achieved. Subsequently, CNIs were withdrawn. In CNI-reduction-Group (group 2), CNI target trough levels were reduced by 40%. In both groups MMF was continued and trough level adjusted (1.5-4 microg/mL). RESULTS: Patients demographics and survival (mean follow-up time: 16.7+/-9 months) was equal (100%). Renal function improved significantly after complete CNI withdrawal while remaining unchanged with CNI-reduction (Creatinine clearance after 12 months: 53+/-24 mg/dL [group 1] vs. 38+/-20 mg/dL [group 2], P=0.01). End-stage renal failure (hemodialysis) was avoided by CNI-withdrawal and occurred only after CNI reduction (n=6; P=0.01). Acute rejection episodes were more common in group 2 (4 vs. 2). Graft function remained stable (echocardiography) within both groups. Adverse events were more common in group 1 (65%) than in group 2 (n=40%) and were responsible for discontinuation in 4 and 0 cases, respectively. CONCLUSIONS: Conversion toward a CNI-free immunosuppression (Mycophenolate, sirolimus) is superior to CNI-reduced immunosuppression in improving renal failure in late HTx recipients. However, this benefit is relativized by the increased incidence and severity of sirolimus/MMF-associated side effects. PMID- 19295319 TI - Postshunt hepatic encephalopathy in liver transplant recipients. AB - BACKGROUND: Preexisting spontaneous portosystemic shunts increase the risk of posttransplantation portal vein thrombosis. Portosystemic shunts may also be placed surgically to manage posttransplant portal vein stenosis/thrombosis. Both types may be complicated by hepatic encephalopathy. METHODS: The database of a major tertiary medical center from 1999 to 2006 was searched for liver transplant recipients with hepatic encephalopathy and stable liver function. The medical and imaging files were reviewed for risk factors, management, and outcome. RESULTS: Of the 244 patients who underwent liver transplantation during the study period, four (1.6%) met the inclusion criteria. Median age at transplantation was 49 years (range 39-54); median time to the first episode of hepatic encephalopathy after transplantation was 23 months (range 2-40). In two patients, a distal splenorenal shunt placed at 1 and 7 months after transplantation to treat portal vein thrombosis led to hepatic encephalopathy at 1 and 33 months later. Both responded to medical therapy. The other two patients had spontaneous splenorenal shunts, and hepatic encephalopathy appeared 33 months and 12 months after transplantation. Treatment consisted of transhepatic percutaneous portal vein dilatation with stent insertion in the first patient and interposition of a venous graft between the superior mesenteric and left intrahepatic portal veins to reroute splanchnic flow in the second patient. CONCLUSIONS: Portosystemic shunts in liver transplant recipients with stable graft function may be associated with hepatic encephalopathy. Pretransplant assessment to detect unknown spontaneous shunts is important. Restoration of portal flow is the preferred procedure in this setting. PMID- 19295320 TI - Successful tapering of immunosuppression to low-dose monotherapy steroids after living-related human leukocyte antigen-identical renal transplantation. AB - BACKGROUND: Living-related (LR) human leukocyte antigen (HLA)-identical renal transplant (RTx) recipients often receive standard immunosuppression, despite the absence of mismatched major HLA-antigens and the known complications of long-term use of immunosuppression. No data are available on the need for immunosuppression for these specific patients. We wondered whether their immunosuppressive load could be radically reduced. METHOD: Between November 1982 and November 2005, 83 LR HLA-identical RTx were performed in our center. Their unadjusted graft survival was 74% at 10 years. In 29 patients (median time after transplantation 5.6 [range 1.0-21.4] years) with stable uncompromised renal function, we tapered their immunosuppression from triple or dual therapy to prednisolone 5 mg/day. Follow up on prednisolone monotherapy was at least 24 months. RESULTS: In 27 of 29 patients reduction of immunosuppression to prednisolone monotherapy was uneventful. One patient, using dual therapy, developed JC-virus nephropathy resulting in graft loss. One refused further discontinuation of his medication. Four (15%) of the 27 patients on monotherapy developed biopsy-proven recurrence of their original disease. Only one of them showed a transient decline in renal function. One additional patient developed minor proteinuria and a rise in serum creatinine level, as a result of chronic urinary tract infections. The remaining 23 of 27 patients (85%) had an uneventful follow up during 24 months prednisolone monotherapy. CONCLUSION: We conclude that HLA-identical LR RTx recipients who are at least 1 year after transplantation might be treated with low-dose steroid monotherapy. Close surveillance of patients for recurrence of their original disease is recommended to allow for potential early therapeutic intervention. PMID- 19295321 TI - Laparoscopic donor nephrectomy: a plea for the right-sided approach. AB - BACKGROUND: Laparoscopic donor nephrectomy (LDN) has become the preferred procedure for live donor nephrectomy. Most transplant surgeons are reluctant toward right-sided LDN (R-LDN) fearing short vessels and renal vein thrombosis. METHODS: In our institution, selection of the appropriate kidney for donation was based on the same criteria that traditionally governed open donor nephrectomy. All intraoperative and postoperative data were prospectively recorded. RESULTS: One hundred fifty-nine R-LDNs (56%) and 124 left-sided LDNs (L-LDN, 44%) were performed. Demographics did not significantly differ. Complications occurred in 10 (6%) vs. 23 (19%) procedures (R-LDN vs. L-LDN, P=0.002), resulting in 2 and 11 conversions, respectively. Right-sided kidney donation was the only independent preventative factor for complications in multivariate analysis (P=0.008, Odds ratio 0.33). R-LDN was associated with shorter operation time (mean 202 vs. 247 min, P<0.001) and less blood loss (139 vs. 294 mL, P<0.001). Hospital stay was 3 days in both groups. With regard to the recipients, the second warm ischemia time was similar (29 vs. 28 min, P=0.699). CONCLUSIONS: R-LDN is faster and safer than L-LDN and does not adversely affect graft function. R-LDN may be advocated to allow donors to benefit from the advantages of laparoscopic surgery. PMID- 19295322 TI - Outcomes of living-related liver transplantation for Wilson's disease: a single center experience in China. AB - BACKGROUND: Although orthotopic liver transplantation provides a therapeutic option for patients with Wilson's disease (WD) presenting fulminant liver failure or drug resistance, it is still unclear whether the living-related liver transplantation (LRLT) can result in long-term therapeutic effect on WD. METHODS: Here, we report a retrospective analysis of LRLT for 36 cases of WD patients. The indications for LRLT were fulminant hepatic failure in two patients and chronic advanced liver disease in 32 patients including 13 patients with Wilsonian neurologic manifestations. Two patients presented with severe Wilsonian neurologic manifestations even though their liver functions were stable. RESULTS: Results revealed that the survival of posttransplant patients or grafts at 1, 3, and 5 years was 91.7%, 83.3%, 75%, or 86.1%, 77.8%, 75%, respectively. Pretransplant intensive care unit-bound and model for end-stage liver disease score were indicated as independent factors predictive of patient survival. Patients with neurologic abnormalities showed significant improvement after liver transplant. CONCLUSION: Our results indicate LRLT is an excellent therapeutic modality for WD patients with end-stage liver disease. Better pretransplant conditions appeared to be advantageous in gaining better survival outcomes of patients undergoing LRLT. PMID- 19295323 TI - Timing of Epstein-Barr virus acquisition and the course of posttransplantation lymphoproliferative disorder in children. AB - BACKGROUND: To investigate the clinical course and risk factors of Epstein-Barr virus (EBV)-associated posttransplantation lymphoproliferative disorder (PTLD) after liver transplantation (LT) among children in Taiwan where children acquired EBV infection in early childhood. METHODS: In the retrospective study, 67 children underwent LT in our hospital and survived for more than 3 months were recruited. Various predisposing risk factors, including viral status, nutritional status, age at transplantation, and medications, were assessed. The diagnosis of EBV-associated PTLD in these liver transplanted patients was confirmed by histologic examination. RESULTS: Eight children developed EBV-associated PTLD after LT, and all (100%) had gastrointestinal tract mucosa-associated lymphoid tissue involvement with the initial presentation as bloody stool. The incidence of PTLD is 11.9% (8/67) in the liver transplanted children in our hospital. Children who received LT before 1 year of age had a higher risk of EBV-associated PTLD than others (relative risk [RR]=10.37, P=0.006). The absence of EBV nuclear antigen antibody in recipients before LT also increased the risk (RR=8.63, P=0.018). The RR of EBV-associated PTLD increased to 13.3 (P=0.002) in EBV naive children who received LT before 1 year of age. CONCLUSIONS: Additional risk of EBV naivity and transplantation age below 1 year was evident in our series to the development of EBV-associated PTLD. Acquired EBV infection in early infancy after the LT may increase the risk of gastrointestinal tract involvement of EBV associated PTLD in Taiwan. PMID- 19295324 TI - Prediction of cardiac complications after liver transplantation. AB - BACKGROUND: Orthotopic liver transplantation (OLT) stresses the cardiovascular system, and cardiac complications after OLT are common. METHODS: Hundred ninety seven patients (>or=40 years) who had OLT from 2002 to 2007 were reviewed to identify predictors of cardiac complications within 6 months after transplantation. RESULTS: Median age was 56 years (40-75 years); 69% men. Reasons for OLT were hepatitis C virus (HCV) 45.5%, alcohol 22%, hepatocellular carcinoma (HCC) 8%, primary biliary cirrhosis 10%, and others 14.5%. Eighty-two patients suffered one or more cardiac complications within 6 months after OLT (pulmonary edema=61 [overt heart failure=7], arrhythmia=13, pulmonary hypertension=7, pericardial effusion=2, and right atrial thrombus=1). Cardiac causes were the leading cause of death (n=5; 23.8% of all mortality). By multivariate analysis, after adjusting for age and sex, independent predictors were adverse intraoperative cardiovascular events (adjusted odds ratio; 95% confidence interval: 5.89, 1.82-19.14), history of cardiac disease (2.42, 0.89-6.6), and i MELD (integrated model for end-stage liver disease) score (1.08, 1.02-1.14), whereas adverse intraoperative cardiovascular events (5.73, 1.96-16.78) and i MELD (1.07, 1.01-1.13) predicted pulmonary edema. None of the following variables predicted complications: age, sex, OLT indication, body mass index, blood pressure, alcohol and smoking history, pre-OLT investigations (chest X-ray, electrocardiogram, echocardiography, coronary angiography, pulmonary arterial pressure, and 2-methoxy isobutyl isonitrile scan), immunosuppressive treatment, or intraoperative variables (transfusion amount, cadaveric vs. living graft or cold ischemia and rewarming times). CONCLUSIONS: Cardiac complications after OLT are common and were the leading cause of death after surgery. Adverse intraoperative cardiovascular events, previous cardiac disease, and advanced liver disease as quantified by i-MELD score predicted postoperative cardiac complications. PMID- 19295325 TI - The efficacy and tolerability of ezetimibe in cardiac transplant recipients taking cyclosporin. AB - BACKGROUND: Despite statin treatment, hyperlipidemia remains problematic after cardiac transplantation and is associated with the development of cardiac allograft vasculopathy. The cholesterol absorption inhibitor ezetimibe may offer a viable option for add on therapy; however, questions have been raised regarding the safety of this during concomitant cyclosporin treatment. METHODS: This is the first placebo controlled, randomized double blinded trial assessing the efficacy and tolerability of ezetimibe in cardiac transplant recipients receiving cyclosporin. Sixty-eight cardiac transplant patients were randomized to receive ezetimibe (10 mg) or matching placebo for 6 months in addition to usual treatments. Fasting blood tests were performed at regular time intervals during the study. RESULTS: Fifty-nine patients completed the study. At 6 months, ezetimibe had reduced total cholesterol by 18% (5.4+/-1.1 to 4.4+/-0.7 mmol/L, P<0.001), low-density lipoprotein cholesterol by 26% (3.0+/-1.0 to 2.1+/-0.7 mmol/L, P<0.001), and triglycerides by 13.5% (2.3+/-1.3 to 1.8+/-0.9 mmol/L, P=0.02). Tolerability was excellent with no patients experiencing predefined safety endpoints. An equal number of patients withdrew consent from each arm of the study because of perceived side effects. Specific analysis confirmed ezetimibe had no significant effect on cyclosporin levels. CONCLUSION: We conclude that ezetimibe is both efficacious and tolerable in cardiac transplant recipients taking cyclosporin. It can be safely considered as add on therapy in patients taking statins (or as monotherapy) to further reduce low-density lipoprotein levels, which may in turn reduce the risk of cardiac allograft vasculopathy. PMID- 19295326 TI - CD3+-mediated rejection and C4d deposition in two composite tissue (bilateral hand) allograft recipients after induction with alemtuzumab. AB - BACKGROUND: New therapies are being introduced in reconstructive transplant surgery to enhance composite tissue allograft survival. METHODS: Alemtuzumab was used in two bilateral hand allograft recipients to cause lymphocyte depletion. RESULTS: Although profound leukopenia and lymphopenia developed, several episodes of acute rejection occurred both in the early and late posttransplant period. Cell-mediated rejection was diagnosed during acute rejection episodes. Intraluminal C4d deposits were found in the capillaries not only accompanying cellular rejection, but also in the absence of clinical rejection. However, their significance is unclear because donor-specific antibodies were absent, there were no pathological signs of injury, allograft function was not impaired, and clinical signs of rejection resolved. CONCLUSION: These findings suggest that alemtuzumab may not prevent cell-mediated rejection of a hand allograft transplant. Furthermore, C4d deposition warrants attention in clinical composite tissue allotransplants. PMID- 19295327 TI - Pulsatile perfusion preservation of en bloc kidneys from small pediatric donors. PMID- 19295328 TI - Streptococcus equi primary bacteremia in a hepatitis-C cirrhotic patient. PMID- 19295329 TI - Prevalence of drug-resistant HIV-1 in rural areas of Hubei province in the People's Republic of China. AB - OBJECTIVE: To determine the prevalence of drug-resistant HIV-1 and the efficacy of first-line highly active antiretroviral therapy (HAART) regimens consisted of generic nucleoside reverse transcriptase inhibitor and nonnucleoside reverse transcriptase inhibitor among 339 study subjects in rural areas of Hubei province, China. METHODS: Two cross-sectional studies were conducted to investigate 150 HAART-naive (99 received subsequent therapy) between 2003 and 2005 and 288 HAART-experienced patients mainly between 2005 and 2006. Patients' CD4+ T-cell count and viral load were determined. HIV-1 pol gene fragments were amplified from patients' plasma by reverse transcriptase-polymerase chain reaction, subsequently sequenced and analyzed. RESULTS: About 83.5% of the patients were from rural villages. They were dominantly infected with subtype B' HIV-1 (96.7%) through paid blood donation (64.6%) and related blood transfusion (28.3%). We found that there was a steady increase of CD4 count over time among treated patients without detectable viral load (186/288, 64.6%). There was, however, an increasing prevalence of nucleoside reverse transcriptase inhibitor- and nonnucleoside reverse transcriptase inhibitor-resistant mutations among patients with detected viremia (102/288, 35.4%) after treatment for 3-6 (24.3%), 9-12 (57.1%), and 20-24 (63.3%) months, respectively. The increasing rates were associated with significant CD4 count drop and viral load increase. Some patients also developed multidrug-resistant mutants. CONCLUSIONS: : We report the first HIV-1 drug resistance study after 2 years on HAART among Chinese patients living in rural villages. Our data suggest that a significant portion of patients are failing first-line regimens with a trend of AIDS progression. It is therefore necessary to maximize the drug adherence and to make affordable second-line HAART regimens available immediately. Our results have implications for implementing HAART in underresourced developing country settings. PMID- 19295330 TI - Envelope coreceptor tropism, drug resistance, and viral evolution among subtype C HIV-1-infected individuals receiving nonsuppressive antiretroviral therapy. AB - BACKGROUND: In resource-constrained settings, antiretroviral treatment (ART) is often continued based on clinical and CD4 responses, without virologic monitoring. ART with incomplete viral suppression was assessed in 27 subjects with subtype C HIV-1. METHODS: Plasma HIV-1 RNA, drug resistance, viral tropism, and evolution in polymerase (pol) and envelope (env) genes were measured. The association between these viral parameters and CD4 cell change over time was analyzed using linear regression models. RESULTS: Increased area under the curve of HIV-1 RNA replication was a predictor of lower CD4 cell gains (P < 0.007), while less drug resistance measured as a genotypic susceptibility score (GSS) (P = 0.065), and lower rates of evolution in pol and env genes (P = 0.08 and 0.097, respectively) measured as genetic distance were modestly associated with increasing CD4 cell counts. Evolution of pol and env were correlated (R2 = 0.48, P = 0.005), however, greater evolution was identified in env vs. pol (P < 0.05). CXCR4-usage (X4) was detected in 14/27 (52%) but no differences in CD4 cell change or plasma viremia were associated with X4-usage. DISCUSSION: Among subtype C HIV-1 infected patients in Zimbabwe receiving incompletely suppressive ART, higher virus replication and lower CD4 cell gains were associated with drug resistance and evolution of polymerase and envelope. PMID- 19295331 TI - Early control of HIV-1 infection in long-term nonprogressors followed since diagnosis in the ANRS SEROCO/HEMOCO cohort. AB - BACKGROUND: To clarify early correlates and natural history of HIV long-term nonprogressors (LTNPs) since HIV diagnosis. METHODS: Patients enrolled in the French ANRS SEROCO/HEMOCO cohort with CD4 count >500 cells/mm3 at HIV diagnosis. LTNP status was defined as being asymptomatic, antiretroviral free, and with CD4 cell count >500 cells/mm3 for >8 years after HIV diagnosis. In LTNPs, we modeled the biological markers' progression through a joint model. Factors associated with loss of LTNP status were identified through a Cox model. RESULTS: Sixty (9%) of 664 patients were identified as LTNPs during follow-up. At enrollment, HIV RNA was 1.85 log copies/10(6) PBMCs and high HIV DNA increase were associated with an increased risk of losing LTNP status [adjusted hazard ratio: 2.8 (1.2-6.8) and 2.2 (1.0-4.8), respectively]. CONCLUSIONS: LTNP status is established in the first years of HIV infection, low HIV DNA level at enrollment and slow increase of HIV DNA being associated with maintained LTNP status. PMID- 19295332 TI - Health-related quality of life in a randomized trial of antiretroviral therapy for advanced HIV disease. AB - OBJECTIVE: To assess and compare alternative approaches of measuring preference based health-related quality of life (HRQoL) in treatment-experienced HIV patients and evaluate their association with health status and clinical variables. DESIGN: Cross-sectional study. SETTING: Twenty-eight Veterans Affairs hospitals in the United States, 13 hospitals in Canada, and 8 hospitals in the United Kingdom. PATIENTS: Three hundred sixty-eight treatment-experienced HIV infected patients enrolled in the Options in Management with Antiretrovirals randomized trial. MEASUREMENTS: Baseline sociodemographic and clinical indicators and baseline HRQoL using the Medical Outcome Study HIV Health Survey (MOS-HIV), the EQ-5D, the EQ-5D visual analog scale (EQ-5D VAS), the Health Utilities Index Mark 3 (HUI3), and standard gamble (SG) and time trade-off (TTO) techniques. RESULTS: The mean (SD) baseline HRQoL scores were as follows: MOS-HIV physical health summary score 41.70 (11.16), MOS-HIV mental health summary score 44.76 (11.38), EQ-5D 0.77 (0.19), HUI3 0.59 (0.32), EQ-5D VAS 65.94 (21.71), SG 0.75 (0.29), and TTO 0.80 (0.31). Correlations between MOS-HIV summary scores and EQ 5D, EQ-5D VAS, and HUI3 ranged from 0.60 to 0.70; the correlation between EQ-5D and HUI3 was 0.73; and the correlation between SG and TTO was 0.43. Preference based HRQoL scores were related to physical, mental, social, and overall health as measured by MOS-HIV. Concomitant medication use, CD4 cell count, and HIV viral load were related to some instruments' scores. CONCLUSIONS: On average, preference-based HRQoL for treatment-experienced HIV patients was decreased relative to national norms but also highly variable. Health status and clinical variables were related to HRQoL. PMID- 19295333 TI - Successful integration of tuberculosis and HIV treatment in rural South Africa: the Sizonq'oba study. AB - BACKGROUND: Tuberculosis (TB) is the leading cause of death among HIV-infected patients worldwide. In KwaZulu-Natal, South Africa, 80% of TB patients are HIV coinfected, with high treatment default and mortality rates. Integrating TB and HIV care may be an effective strategy for improving outcomes for both diseases. METHODS: Prospective operational research study treating TB/HIV-coinfected patients in rural KwaZulu-Natal with once-daily antiretroviral (ARV) therapy concurrently with TB therapy by home-based, modified directly observed therapy. Patients were followed for 12 months after ARV initiation. RESULTS: Of 119 TB/HIV coinfected patients enrolled, 67 (56%) were female, mean age was 34.0 years, and median CD4 count was 78.5 cells per cubic millimeter. After 12 months on ARVs, mean CD4 count increase was 211 cells per cubic millimeter, and 88% had an undetectable viral load; 84% completed TB treatment. Thirteen patients (11%) died; 10 (77%) with multidrug-resistant or extensively drug-resistant TB. There were few severe adverse events or immune reconstitution events. Adherence was high with 93% of study visits attended and 99% of ARV doses taken. CONCLUSIONS: Integration of TB and HIV treatment in a rural setting using concurrent home based therapy resulted in excellent adherence and TB and HIV outcomes. This model may result in successful management of both diseases in other rural resource-poor settings. PMID- 19295334 TI - Racial differences in changes of metabolic parameters and body composition in antiretroviral therapy-naive persons initiating antiretroviral therapy. AB - OBJECTIVE: To assess by race/ethnicity long-term changes in metabolic parameters and body composition among treatment-naive persons initiating antiretroviral therapy (ART). METHODS: We compared changes in 398 participants (African American n = 243, Latino n = 43, white n = 112) initiating ART. At baseline, 1-month (metabolic parameters only) and 4-month follow-up intervals (anthropometric measurements) were performed and fasting metabolic parameters measured. Rates of change over time and overall mean changes from baseline were compared. RESULTS: Latinos had the greatest increase in glucose and insulin resistance and greatest loss of mid-arm and mid-thigh subcutaneous tissue areas. On average, mid-arm and mid-thigh nonsubcutaneous tissue areas increased in all races. Waist subcutaneous tissue area decreased only for Latinos. Visceral tissue area increased the most for Latinos and whites. For all groups, the initial increase in high-density lipoprotein cholesterol was sustained. The initial increase in low-density lipoprotein cholesterol was followed by a gradual decline in all groups. Triglycerides increased for all groups; the increase being the least for African Americans. CONCLUSIONS: In this prospective long-term evaluation, changes in metabolic parameters and body composition varied across race groups. Latinos experienced the most unfavorable changes. Such changes should be monitored over time as the identified differences may impact ART selection. PMID- 19295335 TI - Cardiovascular complications in HIV management: past, present, and future. AB - Highly active antiretroviral therapy (HAART) has significantly improved the prognosis for many individuals with HIV infection. Consequently, HIV infection has become a chronic and manageable disease. The focus on long-term management of patients with HIV infection has broadened to include comorbid conditions, most notably cardiovascular disease. Patients with HIV infection share many cardiovascular risk factors with the general population, and HIV infection itself may increase cardiovascular risk. Changes in lipid profiles associated with increased cardiovascular risk that have been observed with some HAART regimens have been a cause for concern among clinicians who treat HIV-infected patients. However, the lipid effects of HAART seem to depend on the type and duration of regimens employed. They can be managed effectively according to current guidelines that recommend lifestyle changes (eg, improved diet, increased exercise, smoking cessation) and pharmacologic therapy described in established treatment paradigms for patients on antiretroviral therapy and similar to measures currently used by the general population. A review of the clinical data indicates that the virologic and immunologic benefits of HAART clearly outweigh any metabolic effects observed in some patients over time and that preexisting, established cardiovascular risk factors contribute significantly to the potential development of cardiovascular events. These benefits of antiretroviral therapy have been demonstrated in studies comparing the superior efficacy of continuous vs. intermittent HAART. PMID- 19295336 TI - Viral load and CD4+ T-cell dynamics in primary HIV-1 subtype C infection. AB - BACKGROUND: Most knowledge of primary HIV-1 infection is based on subtype B studies, whereas the evolution of viral parameters in the early phase of HIV-1 subtype C infection is not well characterized. METHODS: The kinetics of viral RNA, proviral DNA, CD4+ T-cell count, and subsets of CD4+ T cells expressing CCR5 or CXCR4 were characterized in 8 acute and 62 recent subtype C infections over the first year postseroconversion. RESULTS: The viral RNA peak was 6.25 +/- 0.92 log10 copies per milliliter. After seroconversion, heterogeneity among acute cases was evident by patterns of change in viral load and CD4+ T-cell count over time. The patterns were supported by the rate of viral RNA decline from peak (P = 0.022), viral RNA means (P = 0.005), CD4 levels (P < 0.001), and CD4 decline to 350 (P = 0.011) or 200 (P = 0.046). Proviral DNA had no apparent peak and its mean was 2.59 +/- 0.69 log10 per 106 peripheral blood mononuclear cell. In recent infections, viral RNA set point was 4.00 +/- 0.97 log10 and viral RNA correlated inversely with CD4+ T cells (P < 0.001) and directly with proviral DNA (P < 0.001). CONCLUSIONS: Distinct patterns of viral RNA evolution may exist shortly after seroconversion in HIV-1 subtype C infection. The study provides better understanding of the early phase of subtype C infection. PMID- 19295337 TI - Preexposure antiretroviral prophylaxis attitudes in high-risk Boston area men who report having sex with men: limited knowledge and experience but potential for increased utilization after education. AB - BACKGROUND: Preexposure prophylaxis (PrEP) could protect individuals engaging in repeated high-risk behaviors from HIV infection. Understanding the demographic and behavioral predictors of intent-to-use PrEP may prove useful to identify clinical trial participants. METHODS: In 2007, 227 HIV-uninfected men who report having sex with men (MSM) recruited through modified respondent-driven sampling completed an interviewer-administered survey assessing prior PrEP use and awareness, future intent-to-use PrEP, demographics, sexual risk, psychosocial variables, and drug/alcohol use. Bivariate and multivariable logistic regression procedures examined predictors of intent-to-use PrEP. RESULTS: Mean age of participants was 41 (SD = 9.1); 54% were nonwhite. One participant reported prior off-label PrEP use (medication obtained from his HIV-infected brother). Nineteen percent had previously heard of PrEP, whereas 74% reported intent-to-use PrEP if available after being educated about its potential. In multivariable analysis controlling for age and race/ethnicity, significant predictors of intent-to-use PrEP included the following: less education [odds ratio (OR) = 7.7; P = 0.04], moderate income (OR = 13.0; P = 0.04), no perceived side effects from taking PrEP (OR = 3.5; P = 0.001), and not having to pay for PrEP (OR = 4.2; P = 0.05). DISCUSSION: Many New England MSM indicated an interest in using PrEP after learning about its potential, particularly if they could obtain PrEP at no expense and if PrEP had no side effects. Less educated MSM and those who knew less about PrEP and antiretroviral therapy before entering the study were more open to using antiretroviral therapy for prevention once they had received some information suggesting its potential value. Findings suggest that careful educational messages are necessary to ensure appropriate PrEP use if clinical trials reveal partial efficacy. PMID- 19295338 TI - High frequency of HIV-1 dual infections among HIV-positive individuals in Cameroon, West Central Africa. AB - OBJECTIVES: To determine the frequency of dual inter- and intra-subtype HIV-1 infection among a cohort of 64 longitudinally-studied, HIV-1-positive individuals in Yaounde, Cameroon. METHODS: Blood was collected every 3-6 months for up to 36 months and RNA was extracted from plasma. Gag fragment (HxB2 location 1577-2040) was amplified by nested RT-PCR, and mixed-time-point Heteroduplex Assays (HDAs) were performed. As heteroduplexes in this assay indicate >or=5% genetic discordance in the gag fragment, their presence reveals dual infection. Results were confirmed by phylogenetic analysis. RESULTS: Heteroduplexes were generated by specimens of 10 subjects (15.6%). Kaplan-Meier nonparametric estimate of maintenance of single infection was calculated; the rate/year of a 2 infection was found to be approximately 11%. Dual infection was identified in the final specimens of five subjects, after as much as 18 months follow-up, while for the remaining five subjects, dual infection was identified in interim specimens within an average of 10 months follow-up. Analysis of samples obtained after dual infection from each of these latter five subjects revealed two patterns: reversion to initial strain, or replacement of initial strain. Four subjects were dually-infected with HIV-1 strains of the same subtype, while 6 were infected with different subtypes. CONCLUSIONS: The high prevalence of recombinant HIV-1 strains in Cameroon may in part be explained by the high frequency of dual infection. In this genetically-diverse HIV-1 milieu, dual infections and the recombinant viruses they generate are strongly driving viral evolution, complicating vaccine strategies. PMID- 19295339 TI - Crack-cocaine use accelerates HIV disease progression in a cohort of HIV-positive drug users. AB - BACKGROUND: HIV infection is prevalent among substance abusers. The effects of specific illicit drugs on HIV disease progression have not been established. We evaluated the relationship between substances of abuse and HIV disease progression in a cohort of HIV-1-positive active drug users. METHODS: A prospective, 30-month, longitudinal study was conducted on 222 HIV-1 seropositive drug users in Miami, FL. History of illicit drug, alcohol, and medication use, CD4+ cell count, and viral load were performed every 6 months. RESULTS: Crack cocaine users were 2.14 times [95% confidence interval (CI): 1.08 to 4.25, P = 0.029] more likely to present a decline of CD4 to 50 copies/mL) was examined using a staged multivariable modeling approach. RESULTS: Among 1221 participants, a broad distribution of appointment nonadherence was observed, with 40% of patients missing at least 1 in every 4 scheduled visits. The adjusted odds of appointment nonadherence were 1.85 times higher in African American patients compared with whites [95% confidence interval (CI) = 1.61 to 2.14]. Appointment nonadherence was associated with virologic failure (odds ratio = 1.78, 95% CI = 1.48 to 2.13) and partially mediated the relationship between African American race and virologic failure. African Americans had 1.56 times the adjusted odds of virologic failure (95% CI = 1.19 to 2.05), which declined to 1.30 (95% CI = 0.98 to 1.72) when controlling for appointment nonadherence, a hypothesized mediator. CONCLUSIONS: Appointment nonadherence was more common in African American patients, associated with virologic failure, and seemed to explain part of observed racial disparities in virologic failure. PMID- 19295366 TI - Iris cross-sectional area decreases with pupil dilation and its dynamic behavior is a risk factor in angle closure. AB - PURPOSE: To estimate the change in iris cross-sectional (CS) area with pupil dilation using anterior segment optical coherence tomography comparing eyes with angle closure (AC) to open angle glaucoma (OAG). METHODS: Sixty-five patients from the Wilmer Glaucoma service, 36 with definite or suspected OAG and 29 with definite or suspected AC, underwent anterior segment optical coherence tomography imaging under 3 conditions (pupil constriction to light, physiologic dilation in the dark, and after pharmacologic dilation). The nasal and temporal iris CS areas were measured with custom software, 3 times in each of 4 meridians. The principal outcome variables were iris CS area and change in iris CS area/mm pupil diameter change. The relation of these parameters to potential variables that would influence iris area was estimated by multivariate regression. RESULTS: CS area was smaller in eyes with larger pupil diameter, those that had undergone trabeculectomy, and those of European-derived persons (P<0.05 for all in a univariate analysis). In a multivariate model with CS area as the dependent variable, larger pupil diameter (with a 0.19 mm decrease in CS area for each 1 mm of pupil enlargement, P=0.0002), and trabeculectomy remained significant factors. In a second multivariate model, AC irides had less change in CS area/mm pupil enlargement than OAG or OAG suspects (P=0.01). Change in iris CS area was essentially complete in 5 seconds (n=10 eyes). CONCLUSIONS: The iris loses nearly half its volume from a pupil diameter of 3 to 7 mm, probably by eliminating extracellular fluid. Smaller iris CS area change with physiologic pupil dilation is a potential risk factor for AC. Dynamic iris CS area change deserves testing as a prospective indicator of AC. PMID- 19295367 TI - Repeat selective laser trabeculoplasty. AB - PURPOSE: To evaluate the efficacy of repeat 360-degree selective laser trabeculoplasty (SLT) in glaucoma patients with prior successful 360-degree SLT. DESIGN: Retrospective chart review. METHODS: Forty-four eyes of 35 patients, > or =18 years of age, with open-angle glaucoma (primary open-angle, pseudoexfoliation, or pigmentary glaucoma), uncontrolled on maximum tolerable medical therapy, underwent an initial 360-degree SLT (SLT1), which was successful for > or =6 months, but eventually lost efficacy and was followed by a repeat 360 degree SLT (SLT2). Patients with prior argon laser trabeculoplasty or other glaucoma surgery, before or during the study period, were excluded. Intraocular pressure (IOP) measurements were recorded before each procedure and 1 to 4 weeks, 1 to 3 months, and 5 to 8 months posttreatment and 15 to 21 weeks after the initial SLT. RESULTS: Reduction in IOP after SLT1 and SLT2 was significantly less with repeat treatment at 1 to 3 months, with average decreases of -5.0 and -2.9 mm Hg, respectively (P=0.01), but there were no statistically significant differences between treatments at the other equivalent time points. Using a definition of "success" as > or =20% peak IOP reduction, success rates for SLT1 and SLT2 were not significantly different. There was also no significant difference in eyes that received SLT2 6 to 12 months after SLT1 compared with those that received SLT2 12 months or more after SLT1. CONCLUSIONS: Our findings suggest that repeat 360-degree SLT may be safe and effective after an initially successful 360-degree SLT has failed. These results may be achieved as early as 6 months after the first treatment. PMID- 19295368 TI - Temporal evolution of intraocular pressure elevation after pupillary dilation in pigment dispersion syndrome. AB - PURPOSE: To report 4 patients with pigment dispersion syndrome (PDS) who had delayed intraocular pressure (IOP) spikes after pharmacologic pupillary dilation. METHODS: Four patients with a diagnosis of PDS with documented IOP spike after pharmacologic pupillary dilation were included. Study patients were examined before and after pupillary dilation. The amount of pigment present in the anterior chamber and the IOP were measured at hourly intervals. RESULTS: Although maximal pigment liberation occurred immediately after maximal dilation, the IOP continued to elevate for at least 1.5 hours. CONCLUSIONS: The increase in IOP after pupillary dilation may not occur simultaneously with maximal pigment liberation but may follow it after the pigment has settled out of the anterior chamber. This has implications for monitoring patients with PDS after dilation to detect and treat rises in IOP. PMID- 19295369 TI - Artifacts on the optic nerve head analysis of the optical coherence tomography in glaucomatous and nonglaucomatous eyes. AB - PURPOSE: To determine the frequency and type of imaging artifacts in the optic nerve head (ONH) analysis protocol of the optical coherence tomograph (Stratus OCT) in healthy and glaucomatous eyes. DESIGN: Retrospective study. METHODS: Two experienced operators reviewed imaging obtained in 264 eyes from 264 participants (146 controls, 118 glaucoma patients). Each participant had a minimum of 3 scans using fast ONH protocol. Only scans with good quality were included in the study. RESULTS: Thirty-two eyes (12%) had imaging artifacts in at least one of the radial scans distributed as follows: 12 eyes showed incorrect identification of vitreous-retinal surface plane, 3 eyes had incorrect positioning of retinal pigment epithelium-choriocapillaris border reference points, 10 eyes had vitreous tuft and/or remnants of posterior hyaloid artery above the optic disk that were mistakenly included in the estimation of optic disk parameters (rim and cup area), and 7 eyes with small optic disk were reported with rim area of 0 because of erroneous placement of reference planes. CONCLUSIONS: In the Stratus OCT ONH protocol, careful inspection of each of the 6 radial scans is needed, as artifacts may be present in any of these scans. For the most common artifact found, modifications on retinal sensitivity may be successful in correcting such errors. PMID- 19295370 TI - A comparative study between diode laser cyclophotocoagulation and the Ahmed glaucoma valve implant in neovascular glaucoma: a long-term follow-up. AB - BACKGROUND: To compare the efficacy, safety, and long-term results of intraocular pressure (IOP) reduction by diode laser contact cyclophotocoagulation (DCPC) and Ahmed glaucoma valve (AGV) implant in cases of neovascular glaucoma. METHODS: A total of 66 eyes of 66 patients with neovascular glaucoma were prospectively assigned to either DCPC or AGV implantation. All patients underwent a baseline complete ophthalmologic examination and IOP measurement by Goldmann applanation tonometry before and after 1,3, 6, 12, and 24 months follow-up. Complications and the number of medications were recorded. RESULTS: Mean age of the patients was 60.0+/-11.7 years (range: 20 to 85) in the DCPC group and 57.2+/-10.3 years (range: 20 to 85) in the AGV group. The preoperative IOP was 43.4+/-11.9 mm Hg and 43.3+/-7.4 mm Hg for the DCPC and AGV group, respectively (P>0.05). The postoperative IOP was 16.5+/-11.3 mm Hg and 22.09+/-7.6 mm Hg for the DCPC and AGV groups, respectively (P>0.05) at the last visit. Kaplan-Meier survival analysis showed a probability of success at 24 months of 61.18% and 59.26% for the DCPC and AGV groups (P>0.05). All the patients had a visual acuity of hand movement or worse preoperatively. Visual acuity decreased in 6 eyes (24%) in the DCPC group and 9 eyes (27%) in AGV group. Complications included anterior segment inflammation in 5 eyes (20%), neurotrophic keratitis in 2 eyes (8%), and hypotony in 3 eyes (15%) in the DCPC group and hyphema in 5 eyes (15%) and tube occlusion in 3 eyes (9%) in AGV group. CONCLUSIONS: There was no significant difference in the success rate between the DCPC and AGV implantation in neovascular glaucoma treatment. However, DCPC is less time consuming and easier method for lowering IOP in patients with neovascular glaucoma. PMID- 19295371 TI - Immunohistochemical expression of HLA-DR in the conjunctiva of patients under topical prostaglandin analogs treatment. AB - PURPOSE: Subclinical inflammation may be observed in patients using topical antiglaucomatous drugs. The objective of this study was to investigate inflammation in conjunctiva of glaucoma patients using prostaglandin analogs, by the detection of an immunogenetic marker (HLA-DR) and compare the effect of 3 different drugs: latanoprost, bimatoprost, and travoprost in the induction of this inflammation. SUBJECTS AND METHODS: Thirty-three patients with primary open angle glaucoma were evaluated without and with prostaglandin analogs topical therapy. Imprints of conjunctival cells were obtained, fixed on glass slides, and prepared for immunohistochemical analysis. RESULTS: Before the use of prostaglandin analogs, 4 of the 33 patients evaluated presented expression of HLA DR in the conjunctiva (mild). After 1 month on prostaglandin analog treatment, all but 1 patient presented HLA-DR staining. HLA-DR expression of these 32 patients was scored as mild (19 patients), medium (11 patients), or intense (2 patients). The differences were statistically significant both when the presence and the increased expression of HLA-DR were considered (P<0.001). When the 3 different groups were analyzed (latanoprost, bimatoprost, and travoprost) no statistically significant difference was found (P=0.27). CONCLUSIONS: The use of prostaglandin analogs eye drops provokes a subclinical inflammatory reaction, observed by HLA-DR expression, even after a short period of treatment, independently of the class of the prostaglandin analogs used. PMID- 19295372 TI - Trends in rates of primary angle closure glaucoma and cataract surgery in England from 1968 to 2004. AB - AIM: Eyes that are predisposed to primary angle closure usually have a shallow anterior chamber secondary to a relatively forward position of the lens and progressive lens thickening with ageing. The aim of this study was to examine trends over time in rates of primary angle closure glaucoma (PACG) in England, and to compare these rates with rates of cataract surgery. METHODS: Hospital episode statistics and the hospital inpatient inquiry were analyzed for PACG as the main diagnosis between the years 1968 and 2004, and for cataract surgery over the same period. Age-specific and sex-specific rates of PACG and cataract surgery were calculated over 3 representative time periods. RESULTS: Annual rates of patients with PACG did not change significantly from the late 1960s to the mid 1980s and then increased until the early 1990s before reaching a plateau; from 1999 to 2004, rates of patients with PACG declined significantly. From the 1980s to 2004, annual rates of patients undergoing cataract surgery increased significantly and substantially. In the recent period of decline in PACG, the decline was greatest in older age groups, whereas rates of cataract surgery increased significantly in all age groups for both men and women throughout the whole time period. CONCLUSIONS: Rates of patients with PACG have started to decline in recent years, after a long period of increases in rates of patients undergoing cataract surgery. Although other explanations are possible, this lends support to the hypothesis that cataract surgery may reduce the likelihood of acute angle closure. PMID- 19295373 TI - Thickness of the retinal nerve fibre layer estimated with the scanning laser polarimeter in healthy children. AB - PURPOSE: The scanning laser polarimeter [GDx variable corneal compensation (VCC)] measures the thickness of the retinal nerve fibre layer (RNFL) and has, therefore, a possible potential for the diagnosis and follow-up of glaucoma in children. The aim of the present study is to establish reference values of the RNFL thickness in children. METHODS: The RNFL thickness was measured with a GDx VCC in 72 children aged 4 to 15 years without ocular diseases. The difference in the temporal-superior-nasal-inferior-temporal (TSNIT) average (TA), superior average (SA), and inferior average (IA) between the boys and the girls was estimated with a t test. A 95% confidence interval (CI) of the TA, SA, and IA was calculated for the whole group and compared with those of 4 previous studies. The reproducibility of the examination was estimated in 10 consecutive children and was expressed as 95% CI of the differences between the repeated measurements. RESULTS: There were no statistically significant differences in the TA, SA, and IA between the boys and girls. The 95% CI for the TA, SA, and IA was 61.1+/-1.5, 73.5+/-2.3, and 73.5+/-2.0 microm, respectively. These values did not overlap with those reported previously in the adult groups, confirming a statistically significant age-related thinning of the RNFL. The 95% CI of the differences between the repeated measurements was -0.4+/-1.4 microm for TA, -1.5+/-3.6 microm for SA, and 1.6+/-3.9 microm for IA, suggesting that the examination was reproducible. CONCLUSIONS: The GDx VCC examination is reproducible in children. The present RNFL results should represent useful reference values in the evaluation of pediatric glaucoma. PMID- 19295374 TI - Frequency of spontaneous pulsations of the central retinal vein in glaucoma. AB - PURPOSE: To assess the prevalence of spontaneous pulsations of the central retinal vein in eyes with chronic open-angle glaucoma versus normal eyes in the setting of a general ophthalmic practice. METHODS: The clinical observational case-series study included 167 eyes of 84 subjects with chronic open-angle glaucoma and 704 eyes (352 subjects) of a control group. The optic disc was ophthalmoscopically examined using a noncontact ophthalmoscopic lens at the slit lamp in medical mydriasis. RESULTS: Spontaneous pulsations of the central retinal vein were observed in significantly (P=0.005; odds ratio, 0.59; 95% confidence interval, 0.41-0.84) fewer eyes of the glaucoma group (107/167 or 64.1%+/-3.7%%) than in the eyes of the control group (530/704 or 75.3%+/-1.6%). In both groups, the presence of a spontaneous central retinal vein pulsation was statistically (P>0.10) independent of age, sex, refractive error, and intraocular pressure. CONCLUSIONS: As examined in a nonreferral ophthalmic practice, eyes with chronic open-angle glaucoma compared with normal eyes show a significantly lower frequency of detected spontaneous pulsations of the central retinal vein. Confirming previous hospital-based studies, the results suggest that assessment of the central retinal vessel pulsations, as part of an optic disc examination may clinically be useful for the diagnosis of glaucoma. PMID- 19295375 TI - Signal strength is an important determinant of accuracy of nerve fiber layer thickness measurement by optical coherence tomography. AB - PURPOSE: To investigate the effect of signal strength on the measurement of the retinal nerve fiber layer (RNFL) using optical coherence tomography (OCT). METHODS: Eyes with known or suspected glaucoma or nonglaucomatous optic atrophy were scanned twice within the same visit using Stratus OCT's Fast Nerve Fiber Layer Thickness protocol. Only those eyes with 2 high-quality scans (signal strengths of at least 5 and different from each other, no error messages, and no obvious segmentation errors) were included in the study. The RNFL thickness measurements from the initial and the repeat scans were compared and then correlated with the differences in signal strength. Subgroup analyses were performed similarly among patients with average RNFL thickness less than 90 microm and those with at least 90 microm. RESULTS: Scans with higher signal strengths are associated with greater RNFL thickness measurements if the signal strength is less than 7. Scans with signal strength of at least 7 have higher reproducibility. This is true among all patients and subgroups divided on the basis of average RNFL thickness. Additionally, we found that the greater the variability between the initial and repeat scans, the greater the variability in the RNFL thickness measurements. Scans with higher signal strengths have less variability, especially when the optic nerve is relatively healthy. CONCLUSIONS: When measuring the RNFL thickness with the Stratus OCT, it is important to aim for a signal strength of at least 7. Visual field testing may be more reliable in some patients, especially when the optic nerve is significantly compromised. PMID- 19295376 TI - Correlation between the indiana bleb appearance grading scale and intraocular pressure after phacotrabeculectomy. AB - PURPOSE: To investigate the relationship between bleb morphology, recorded using the Indiana Bleb Appearance Grading Scale (IBAGS), and intraocular pressure (IOP) after phacotrabeculectomy. METHODS: Two years postphacotrabeculectomy, a single observer compared bleb morphology to the IBAGS standard photographs in 76 eyes of 76 patients. In addition, the presence or absence of microcysts was recorded. IOP was also measured. RESULTS: On multivariate analysis increasing bleb height was associated with a low IOP (P=0.017). An increase in IBAGS height score by 1 U resulted in a reduction in IOP of 2.16 mm Hg (95% confidence interval=0.40-3.92 mm Hg). In this study, there was no association between vascularity, bleb extent or microcysts, and IOP. There were no cases of bleb leak in this series. CONCLUSIONS: Two years postphacotrabeculectomy increased bleb height, as measured by the IBAGS, was associated with a decrease in IOP, with a 1 point increase in IBAGS height score resulting in a decrease of 2.16 mm Hg. We found no association between bleb extent, vascularity, or the presence or absence of conjunctival microcysts. As there were no cases of bleb leak in this study this characteristic could not be evaluated. PMID- 19295377 TI - A thin honeycomb-patterned film as an adhesion barrier in an animal model of glaucoma filtration surgery. AB - PURPOSE: To evaluate the effectiveness and safety of a thin honeycomb-patterned biodegradable film for glaucoma filtration surgery in rabbits. METHODS: A 7 microm-thick film made from poly(L-lactide-co-epsilon-caprolactone) was placed in the subconjunctival space in one eye of rabbits, with or without full thickness filtration surgery. The film had a honeycomb-patterned surface that faced the subconjunctival Tenon tissue and the other side was smooth. Filtration surgery was also performed in the fellow eye, which received either no adjunctive treatment or 0.4 mg/mL mitomycin C (MMC; n=6 each). Intraocular pressure (IOP) measurements and bleb evaluations using ultrasound biomicroscopy were performed periodically for 28 days after surgery followed by histologic observation. RESULTS: Postoperative IOPs of the film-treated eyes were significantly lower than that of control eyes from day 10 to day 28 (P<0.05), but were not significantly different from those of MMC-treated eyes. The subconjunctival filtration space, detected by ultrasound biomicroscopy, disappeared in 5 control eyes, 1 MMC-treated eye, but none of the film-treated eyes. A bleb leak occurred postoperatively in 2 MMC-treated eyes. Histologically, in eyes without filtration surgery, fibrotic tissue with the film partly attached to it was noted on the honeycomb side, but was minimal on the sclera that faced the smooth side of the film. In eyes with filtration surgery, the honeycomb-patterned film lined the inner bleb wall with minimal inflammatory reaction. CONCLUSIONS: The thin honeycomb-patterned film that attached to the inner bleb wall worked as an adhesion barrier in glaucoma filtration surgery in rabbits, which is worthy of further investigation. PMID- 19295378 TI - Sensitivity and specificity with the glaucoma probability score in Heidelberg Retina Tomograph II in Japanese eyes. AB - PURPOSE: To evaluate the sensitivity and specificity with glaucoma probability score (GPS), a new glaucoma diagnosis program in Heidelberg Retina Tomograph II ver 3.0 in detecting early glaucoma in Japanese eyes. PATIENTS AND METHODS: Sensitivity and specificity with GPS were determined in 148 early stage open angle glaucoma eyes of 148 hospital-based patients (mean deviation >-5 dB) and age-matched and refraction-matched 148 eyes of 148 ophthalmologically normal subjects selected from a population-based sample of the Tajimi Study. The results with GPS were compared with those with Moorfields regression analysis (MRA) and FS Mikelberg discriminant function (FSM). Analysis was also carried out by stratifying the eyes by refraction and disc area. RESULTS: For all glaucoma eyes, sensitivity with GPS (71.7%) was similar to MRA and FSM (75.2%, P=0.8; 77.2%, P=0.5; Fisher exact test). Specificity with GPS (80.7%) was also statistically equivalent with MRA and FSM (91.1%, P=0.06; 87.4%, P=0.3). Although sensitivity with GPS was lower in higher myopic eyes than in the less myopic eyes, specificity did not differ with refractive error. Specificity with GPS was lower and sensitivity higher (P<0.0001) in larger discs (>2.0 mm) than in smaller discs (< or =2 mm). In larger discs, specificity with GPS was lower (P=0.005) than with MRA and in smaller discs, sensitivity with GPS was lower than with MRA or FSM. CONCLUSIONS: In Japanese eyes, sensitivity and specificity with GPS to detect early glaucoma were similar with MRA or FSM, although the specificity tended to be lower than results in white eyes. Subgroup analysis revealed a notable dependency in diagnostic ability on disc area and refractive error. The diagnostic ability of GPS is still not sufficient for glaucoma detection in the Japanese population requiring further investigation. PMID- 19295379 TI - Efficacy and tolerability of nonpenetrating filtering surgery with and without implant in treatment of open angle glaucoma: a quantitative evaluation of the evidence. AB - OBJECTIVE: To evaluate the efficacy and tolerability of nonpenetrating filtering surgery augmented with implant (NPFSI) compared with nonpenetrating filtering surgery without implant (NPFS) in the treatment of patients with open angle glaucoma. METHODS: Pertinent studies were selected through extensive searches of the Cochrane Library, PubMed, Embase, and American Association of Ophthalmology (AAO) and Association for Research in Vision and Ophthalmology (ARVO) meeting abstracts. Seven controlled clinical trials meeting the predefined criteria were systematically reviewed by meta-analysis. The main outcome measures were percentage intraocular pressure (IOP) reduction, complete success rates, and qualified success rates. The pooled estimates were carried out in RevMan version 4.2 software. RESULTS: The weighted mean difference of the percentage IOP reduction from baseline were 6.7% (95% confidence intervals: -3.8 to 17.2) when comparing NPFSI with NPFS. NPFSI was associated with numerically greater, but nonsignificant, IOP lowering efficacy than NPFS. The pooled risk differences comparing NPFSI with NPFS were 0.13 (-0.02 to 0.28) in complete success rate and 0.09 (-0.04 to 0.21) in qualified success rate. Numerically greater, but nonsignificant, proportions of NPFSI patients than NPFS patients achieved the target end point IOP. NPFSI resulted in the same incidences of postoperative complications as NPFS. CONCLUSIONS: NPFSI was associated with equivalent efficacy to standard NPFS in lowering IOP. Comparable proportions of patients reached the IOP target with NPFSI and NPFS. Both surgeries were well tolerated. PMID- 19295380 TI - Determinants of adherence to glaucoma medical therapy in a long-term patient population. AB - PURPOSE: Estimate patient adherence to glaucoma medications and identify potential determinants of nonadherence. DESIGN: Descriptive study. METHODS: Two hundred patients with open angle glaucoma, ocular hypertension, or glaucoma suspects were interviewed regarding their glaucoma and its treatment and their charts were reviewed. Their ophthalmologist completed a brief assessment form. Drug utilization data were extracted from the provincial drug program database. Patients were defined as adherent if they filled at least 75% of the prescribed medication necessary for their treatment. RESULTS: Data were available for 181 patients. About 62.9% were female and the mean age (+/-SD) was 75.1+/-8.8 years. The mean number of years of glaucoma treatment was 10.7+/-9.3. Self-reported treatment adherence was 88.3%. On the basis of the drug database, the proportion of patients who were adherent to treatment was 71.8%. According to physicians, 74.6% of patients were adherent. Among patients considered by physicians as nonadherent, 71.1% (32/45) were adherent and among patients predicted as adherent, 72.1% (98/136) were adherent. There was no significant difference in adherence according to age, sex, education, and income. However, patients using fewer agents (P=0.041), who were widowed (P=0.041), or who lived alone (P=0.042) were more adherent. Patients using prostaglandins analogs or beta-blockers were more adherent than those using carbonic anhydrase inhibitors (P<0.05). CONCLUSIONS: Fewer medications, use of prostaglandin analogs or beta-blockers, living alone, and being widowed were significantly associated with adherence. Physicians were unable to significantly predict which patients are adherent. PMID- 19295381 TI - Glaucomatous disease in patients with normal pressure hydrocephalus. AB - PURPOSE: To investigate the relative prevalence of glaucomatous disease in patients with normal pressure hydrocephalus (NPH) compared with age-matched non NPH controls with hydrocephalus. DESIGN: Case-control study. PARTICIPANTS: Using International Classification of Diseases (ICD-9) codes, we identified the medical records of 72 NPH cases and 72 age-matched controls with hydrocephalus treated at Stanford University Hospital between 1996 and 2007. METHODS: All available electronic medical records were reviewed. The diagnosis of NPH was verified based on clinical presentation, neuroimaging results, and opening pressure on initial access to cerebrospinal fluid space. Glaucoma status was determined based on history and prior medical, laser, or surgical intraocular pressure-lowering therapy. MAIN OUTCOME MEASURE: The proportion of subjects determined as having glaucomatous disease in cases and controls. RESULTS: The prevalence of glaucoma in NPH cases was estimated to be 18.1% in contrast to 5.6% in controls (chi=5.403, P=0.0201). The average age of NPH cases and non-NPH controls was 75.40+/-13.4 and 73.96+/-9.92 years (mean+/-SD), respectively. CONCLUSIONS: We found the prevalence of a glaucoma diagnosis to be 3-fold greater in patients with NPH as compared with age-matched non-NPH controls with hydrocephalus. These findings suggest the possiblity of an association between these 2 neurodegenerative diseases which should be studied prospectively. PMID- 19295382 TI - Reproducibility of the Heidelberg Retina Tomograph III Glaucoma Probability Score. AB - PURPOSE: To evaluate the intrasession reproducibility of the Heidelberg Retina Tomograph III (HRT III) Glaucoma Probability Score (GPS) and analyze the factors that may affect it. PATIENTS AND METHODS: Twenty normal subjects and 20 glaucomatous patients underwent 10 HRT III examinations of 1 randomly selected eye during the same session. The reproducibility of the GPS and its parameters was assessed by calculating the intraclass correlation coefficients (ICCs) and 95% confidence intervals of the study population as a whole, and of the 2 subgroups. The factors influencing individual reproducibility were evaluated by means of regression analysis. RESULTS: The reproducibility of the GPS parameters (at a confidence level of 95%) was almost perfect in the population as a whole (ICCs 0.964-0.856) and in the normal subjects (ICCs 0.976-0.897), but decreased in the glaucoma patients in whom it ranged from almost perfect to substantial (ICCs 0.922-0.705). The parameters were mainly influenced by image quality, but also by diagnosis and age, although the probability of glaucoma did not seem to depend on these variables. CONCLUSIONS: GPS is a highly reproducible diagnostic tool, especially in the case of "good" or better quality images as evaluated by the HRT software; moreover, the reproducibility of the GPS parameters was significantly better in normal subjects than in glaucoma patients. PMID- 19295383 TI - Comparison of quantitative imaging devices and subjective optic nerve head assessment by general ophthalmologists to differentiate normal from glaucomatous eyes. AB - PURPOSE: To compare the ability of subjective assessment of optic nerve head (ONH) and retinal nerve fiber layer (RNFL) by general ophthalmologists and by a glaucoma expert with objective measurements by optical coherence tomography (Stratus OCT, Carl Zeiss Meditec Inc), confocal scanning laser ophthalmoscope (HRT III; Heidelberg Engineering, Heidelberg, Germany), and scanning laser polarimetry (GDx enhanced corneal compensation; Carl Zeiss Meditec Inc, Dublin, CA) in discriminating glaucomatous and normal eyes. METHODS: Sixty-one glaucomatous and 57 normal eyes of 118 subjects were included in the study. Three independent general ophthalmologists and 1 glaucoma expert evaluated ONH stereophotographs. Receiver operating characteristic curves were constructed for each imaging technique and sensitivity at fixed specificity was estimated. Comparisons of areas under these curves (aROCs) and agreement (k) were determined between stereophoto grading and best parameter from each technique. RESULTS: Best parameter from each technique showed larger aROC (Stratus OCT RNFL=0.92; Stratus OCT ONH vertical integrated area=0.86; Stratus OCT macular thickness=0.82; GDx enhanced corneal compensation=0.91; HRT3 global cup-to-disc area ratio=0.83; HRT3 glaucoma probability score numeric score=0.83) compared with stereophotograph grading by general ophthalmologists (0.80) in separating glaucomatous and normal eyes. Glaucoma expert stereophoto grading provided equal or larger aROC (0.92) than best parameter of each computerized imaging device. Stereophoto evaluated by a glaucoma expert showed better agreement with best parameter of each quantitative imaging technique in classifying eyes either as glaucomatous or normal compared with stereophoto grading by general ophthalmologists. The combination of subjective assessment of the optic disc by general ophthalmologists with RNFL objective parameters improved identification of glaucoma patients in a larger proportion than the combination of these objective parameters with subjective assessment of the optic disc by a glaucoma expert (29.5% vs. 19.7%, respectively). CONCLUSIONS: Diagnostic ability of all imaging techniques showed better performance than subjective assessment of the ONH by general ophthalmologists, but not by a glaucoma expert. Objective RNFL measurements may provide improvement in glaucoma detection when combined with subjective assessment of the optic disc by general ophthalmologists or by a glaucoma expert. PMID- 19295384 TI - Nonstitch suprachoroidal technique for T-flux implantation in deep sclerectomy. AB - A nonstitch technique for suprachoroidal T-flux implantation in deep sclerectomy is described. The foot of the T-flux is introduced into the suprachoroidal space through an incision created just posterior to the scleral spur. As both arms of the T-flux implant are also introduced into the surgically created ostia in the Schlemm's canal, the 3-point fixation makes it unnecessary to secure the implant with a suture. PMID- 19295385 TI - Late-onset bleb leakage. PMID- 19295386 TI - Neuroanesthesiology update. AB - The 2008 literature contained numerous articles of interest to physicians providing perioperative care for neurosurgical or neurologically-impaired patients. In this review, we provide a brief summary of common themes and unique or novel reports. Topics addressed are intracranial hemorrhage, traumatic brain injury, neuropharmacology, neuroprotection, spine surgery, and treatment of carotid artery atherosclerotic disease. PMID- 19295387 TI - Awake craniotomy induces fewer changes in the plasma amino acid profile than craniotomy under general anesthesia. AB - In this prospective, observational, 2-armed study, we compared the plasma amino acid profiles of patients undergoing awake craniotomy to those undergoing craniotomy under general anesthesia. Both experimental groups were also compared with a healthy, age-matched and sex-matched reference group not undergoing surgery. It is our intention to investigate whether plasma amino acid levels provide information about physical and emotional stress, as well as pain during awake craniotomy versus craniotomy under general anesthesia. Both experimental groups received preoperative, perioperative, and postoperative dexamethasone. The plasma levels of 20 amino acids were determined preoperative, perioperative, and postoperatively in all groups and were correlated with subjective markers for pain, stress, and anxiety. In both craniotomy groups, preoperative levels of tryptophan and valine were significantly decreased whereas glutamate, alanine, and arginine were significantly increased relative to the reference group. Throughout time, tryptophan levels were significantly lower in the general anesthesia group versus the awake craniotomy group. The general anesthesia group had a significantly higher phenylalanine/tyrosine ratio, which may suggest higher oxidative stress, than the awake group throughout time. Between experimental groups, a significant increase in large neutral amino acids was found postoperatively in awake craniotomy patients, pain was also less and recovery was faster. A significant difference in mean hospitalization time was also found, with awake craniotomy patients leaving after 4.53+/-2.12 days and general anesthesia patients after 6.17+/-1.62 days; P=0.012. This study demonstrates that awake craniotomy is likely to be physically and emotionally less stressful than general anesthesia and that amino acid profiling holds promise for monitoring postoperative pain and recovery. PMID- 19295388 TI - EEG entropy values during isoflurane, sevoflurane and halothane anesthesia with and without nitrous oxide. AB - We hypothesized that like bispectral index, entropy may be anesthetic agent specific. We carried out a study to assess the entropy values of different anesthetics at equi-minimal alveolar concentrations (MACs) with air and nitrous oxide as carrier gases. Thirty adult patients undergoing spine surgery were randomized to receive halothane, isoflurane, or sevoflurane, in 2 stages, (a) with air/oxygen mixture (2:1) and (b) in nitrous oxide/oxygen (2:1). Heart rate, mean arterial blood pressure, response entropy (RE), and state entropy (SE) were noted at 1.0 and 1.5 MACs for each agent. Statistical analysis was done using the 2-way analysis of variance followed by Bonferroni correction and Student t test for paired data. P value of less than 0.05 were considered significant. The demographics and baseline values of heart rate, mean arterial blood pressure, RE, and SE were comparable. Changing from air/oxygen as carrier gas to 66% nitrous oxide in oxygen resulted in significant increase in both RE and SE at 1.0 MAC for all the agents (P<0.05). Among the agents, it was found that both RE and SE values were significantly higher during halothane anesthesia as compared with sevoflurane and isoflurane (P<0.05). At 1.5 MAC for all agents, after addition of nitrous oxide, there was an insignificant reduction in both RE and SE (P>0.05). Again the values of RE and SE remained high for halothane as compared with isoflurane and sevoflurane. In conclusion, our data suggest a possibility of misinterpretation of anesthetic hypnosis when entropy values increase with addition of nitrous oxide to 1 MAC isoflurane and sevoflurane. PMID- 19295389 TI - Acid-base status and hemodynamic stability during propofol and sevoflurane-based anesthesia in patients undergoing uncomplicated intracranial surgery. AB - Propofol anesthesia may induce metabolic disturbances and sevoflurane anesthesia arterial hypotension. This study compares both techniques regarding acid-base and hemodynamic status during intracranial surgery. Sixty-one patients were randomized into 2 groups according to anesthesia maintenance, a propofol group (n=30), and a sevoflurane group (n=31). The anesthesia protocol including rocuronium and remifentanil infusion was otherwise similar in both groups. Arterial blood samples were drawn every 2 hours during the procedure and upon arrival in the intensive care unit to assess acid-base status. The number of hypotensive and hypertensive events served to assess hemodynamic stability. Metabolic acidosis was more frequent during propofol than sevoflurane anesthesia (7 out of 29 and 1 out of 31, P=0.02). Its severity was linearly correlated with lactate concentration (R=0.32), total dose of propofol (R=0.2), and length of procedure (R=0.28). Hyperlactacidemia was also observed during sevoflurane anesthesia, but without acidosis. Hypertension occurred more frequently during propofol than sevoflurane anesthesia (13 out of 30 vs. 1 out of 31, P<0.001), particularly in patients with a past medical history of hypertension. Higher remifentanil infusion rates reduced the risk of hypertension. Conversely, sevoflurane anesthesia favored arterial hypotension (22 out of 31 vs. 12 out of 30, P=0.015). Preoperative morning administration of antihypertensive medications to patients with a history of arterial hypertension was associated with a low probability of hypertensive events, at the cost of more frequent hypotension. In conclusion, propofol anesthesia for intracranial surgery is more frequently associated with lactic acidosis and hypertension; sevoflurane anesthesia may favor arterial hypotension. PMID- 19295390 TI - A comparison of auditory evoked potentials derived from a monitor integrated module versus standard technique. AB - PURPOSE: Measurement of brain stem auditory evoked potentials (BAEP) and midlatency auditory evoked potentials (MLAEP) using a new monitor integrated module was compared with an established device. The aim of this study was to evaluate if the new system could replace the more inconvenient established technique. MATERIAL AND METHODS: MLAEP and BAEP were obtained from 19 anesthetized male patients using the AEP-Module for Monitor S/5 [GE, Helsinki, Finland (S/5)] and Neuropack 4 mini [Nihon Kohden, Tokyo, Japan (Neuropack)]. Significance of different results was calculated by paired t test. Bias was estimated by Bland-Altman statistics. RESULTS: Latencies of BAEP were significantly shorter and amplitudes of BAEP were significantly higher for Neuropack. Mean latencies of MLAEP (Pa and Nb) were not significantly different (Pa 44.1+/-4.4 ms vs. 41.9+/-5.4 ms/Nb 66.4+/-5.6 ms vs. 62.8+/-6.5 ms), but methods are not interchangeable owing to great variability (Pa -13.16 to 8.94 ms, Nb -19.15 to 11.79 ms). CONCLUSIONS: BAEP recorded by S/5 cannot be used for diagnostic interpretation using generally accepted normal values, but can be used for examining changes during the monitoring period. Mean values of Pa and Nb were not significantly different, but values of the S/5 varied above and below the values of Neuropack such that the measurements could not be used for diagnostic interpretation. However, this did not reduce their usefulness for determining adequate hypnosis. PMID- 19295391 TI - Preoperative anxiety in neurosurgical patients. AB - Anxiety is common in surgical patients, with an incidence of 60% to 92%. There is little information on the incidence and severity of preoperative anxiety in patients scheduled for neurosurgery. The aim of this study was to measure the level of preoperative anxiety in neurosurgical patients and to assess any influencing factors. After the Institutional Review Board approval and informed written consent, 100 patients booked for neurosurgery were interviewed preoperatively. Each patient was asked to grade their preoperative anxiety level on a verbal analog scale, Amsterdam Preoperative Anxiety and Information Scale, and a set of specific anxiety-related questions. The anxiety scores and the responses to the questions were compared between the sex, age, weight, diagnosis, and history of previous surgery. The mean age (+/-SD) was 50+/-13 years. The preoperative diagnosis was tumor (n=64), aneurysm (n=14), and other (n=22). Overall verbal analog scale was 5.2+/-2.7; the score was higher for female (5.8+/ 2.8) than male patients (4.6+/-2.5) (P<0.05). Amsterdam Preoperative Anxiety and Information Scale anxiety and knowledge scores were greater for surgery than for anesthesia. Questionnaire results showed that the most common anxieties were waiting for surgery, physical/mental harm, and results of the operation. In conclusion, our study showed that neurosurgical patients have high levels of anxiety, with a higher incidence in females. There was a moderately high need for information, particularly in patients with a high level of preoperative anxiety. PMID- 19295392 TI - Effects of midazolam on brain injury after transient focal cerebral ischemia in rats*. AB - The benzodiazepine, midazolam, is commonly used for sedation and anesthesia in the operating room and the intensive care unit where there is a risk of cerebral ischemia. We therefore examined its ability to reduce damage subsequent to cerebral ischemia. Male Wistar rats were randomly assigned to a high-dose midazolam group or a matched vehicle group and a lower dose of midazolam group or a matched vehicle group. The rats underwent 90 minutes of middle cerebral artery occlusion. In the midazolam groups, the first dose of midazolam (10 or 25 mg/kg) was given by a 10-minute intravenous infusion before ischemia; a second dose of (1/2) the initial dose (5 or 12.5 mg/kg) was given 1 hour after the onset of ischemia. In the vehicle groups, a similar volume of vehicle was given at the same time intervals. Infarct size, NeuN immunopositive cells in the ischemic penumbral and core regions, and neurologic outcome were determined 7 days after ischemia. Compared with vehicle-treated rats, the higher-dose midazolam (25 mg/kg)-treated rats had a smaller infarct size (93.9+/-63.5 mm vs. 152.0+/-53.7 mm, P<0.05), more NeuN immunopositive cells in the ischemic core region (206.7+/ 211.3/mm vs. 40.0+/-66.3/mm, P<0.01), and better neurologic outcome (P<0.05). Midazolam at a lower dose (10 mg/kg) had no significant effects. Although midazolam generated dose-dependent hemolysis, this hemolysis was transient. Midazolam (25 mg/kg) caused a loss of the righting reflex in rats that lasted until 19.9+/-1.3 min after the injection, the anesthetic dose of midazolam in rats is approximately 100x greater than the anesthetic dose for humans. An anesthetic dose of midazolam in rats reduced neuronal damage and improved neurologic outcome 7 days after focal cerebral ischemia, however, it also caused transient hemolysis. PMID- 19295393 TI - Nimodipine prevents transient cognitive dysfunction after moderate hypoxia in adult mice. AB - BACKGROUND: Cognitive changes associated with moderate hypoxia may be related to the elevation of cytosolic calcium (Ca) levels which may, in turn, affect neurotransmitter synthesis and metabolism. We tested whether treatment with nimodipine (NIMO), an L-type Ca channel blocker, would preserve working memory after hypoxic hypoxia. METHODS: We randomized 157 Swiss-Webster, 30 to 35 g mice (6 to 8 wk) to 6 groups, which were exposed to the following gas mixtures for 1 hour: (1) O2 21%; (2) O2 21% followed by 0.1 mg/kg of subcutaneous NIMO; (3) O2 21% followed by vehicle (60% polyethylene glycol/40% methanol); (4) O2 10%; (5) O2 10% then NIMO; (6) O2 10% then vehicle. The Object Recognition Test (ORT) was given once either on Day 1 or Day 7 to assess changes in short-term memory. ORT exploits the tendency of mice to prefer novel over familiar objects. Two identical objects were placed in an arena for 15 minutes of training. During the testing 1 hour later, one of the objects was replaced by a new object. Recognition Index (RI) was used to compare performance. It is defined as the time spent exploring the novel object divided by the time spent exploring both objects, the novel plus the familiar, and this ratio is converted to a percentage. RI was analyzed with analysis of variance. Tukey Honestly Significant Difference tests were used for post hoc comparisons when appropriate. P values <0.05 were considered significant. RESULTS: RI for the control group was 68.3% (SE+/-3.6%). RI was 53.7% (SE+/-3.8%) for the 10% O2 group on the first posttreatment day. O2 saturation (SpO2) for the hypoxic group was 71.7% (SE+/ 0.5%). By Day 7, RI for the 10% O2 group increased to 64.2% (SE+/-4.7%), which was not significantly different from control. On Day 1, RI was 68.6% (SE+/-5.2%) for hypoxic rodents treated with NIMO. These results were statistically significant. Low RI indicates impaired working memory and high RI indicates intact working memory. These results suggest that NIMO prevented impairment of working memory after moderate hypoxia. CONCLUSIONS: NIMO reverses the disturbance of short-term working memory caused by moderate hypoxia in mice. The results may have implications for cognitive changes linked to Ca homeostasis in the postoperative period. PMID- 19295394 TI - Propofol and ketamine-induced anesthetic depth-dependent decrease of CaMKII phosphorylation levels in rat hippocampus and cortex. AB - Ca/calmodulin-dependent protein kinase II (CaMKII) activation through autophosphorylation at threonine 286 was involved in the modulation of neuronal excitability and neurotransmission. Both propofol and ketamine may affect the intracellular Ca levels through N-methyl-D-aspartate receptors or voltage dependent Ca channels, but they have different mechanisms in general anesthesia. The purpose of this study was to investigate the effects of propofol and ketamine on CaMKII total protein and phosphorylation (p-CaMKII) levels in the brain of rats. We found that both propofol and ketamine could induce a decrease of p CaMKII, not CaMKII total protein, in an anesthetic depth-dependent manner, whereas only ketamine caused a dose (50, 100, and 150 mg/kg)-dependent depression of p-CaMKII in hippocampus and frontal cortex of rats after intraperitoneal injections for 30 minutes. The significant depressions of p-CaMKII started at 5 minutes both in hippocampus and frontal cortex of rats after 100 mg/kg propofol treatment, whereas 100 mg/kg ketamine-induced significant depression of p-CaMKII initiated at 30 minutes in hippocampus and 5 minutes (no reduction observed at 15 min) in frontal cortex of rats. The maximum reduction of p-CaMKII with both drugs was at 60 minutes, and then restored to control level at 240 minutes. In addition, we confirmed the depression of p-CaMKII in hippocampus and frontal cortex of rats after 100 mg/kg of propofol or ketamine treatment for 60 minutes by using immunostaining. These results suggested that decreased p-CaMKII levels correlate with anesthetic depths achieved by propofol and ketamine, which may be related to the effects of propofol and ketamine on central nervous system function and their clinical effect. PMID- 19295395 TI - Clinical evaluation of a new multiparameter neuromonitoring device: measurement of brain tissue oxygen, brain temperature, and intracranial pressure. AB - OBJECTIVE: The study presented evaluated the first clinical use of a new multiparameter catheter measuring intracranial pressure (ICP), partial pressure of brain tissue oxygen (ptiO2), and brain temperature (TBr) (Neurovent PTO). To assess the validity of measured ptiO2 a second probe, which represents the current golden standard of ptiO2 measurement, was implanted (Licox system). METHODS: Thirty patients with indicated invasive measurement of ICP under intensive care unit conditions were included. Using a double lumen bolt, ptiO2 was measured simultaneously with Licox and Neurovent PTO. Ex vivo tests on both probes were conducted independently by the manufacturer of the Neurovent PTO (Raumedic AG, Germany). RESULTS: The average of individual mean ptiO2 measurements showed no relevant differences between the Licox (19.5+/-7.1) and Neurovent multiparameter probe (21.7+/-9.5). Twenty-eight Licox probes out of 30 showed proper functioning over the desired monitoring period. Raumedic multiparameter probes displayed a higher malfunction/handling error frequency (2 device errors, 11 handling errors). A comparison of the ptiO2 data between the Licox and Raumedic systems according to Bland and Altman was possible in 18 out of 30 patients and showed acceptable results (mean difference -1.24 mm Hg; limits of agreement: -25.1 to +22.6 mm Hg). A total of 95.2% of 96,083 recordings was within the calculated limits of agreement. Ex vivo tests of the probes after explantation revealed stable ICP and TBr function of the Raumedic probe. Precision of Zero ptiO2 did not differ between the probes, whereas precision of the 150 mm Hg ptiO2 was greater in the Raumedic probes. CONCLUSIONS: Combining 3 different neuromonitoring functions in 1 probe might ease monitoring by making a second (ptiO2) probe unnecessary. Interpretation of our data is limited by several factors: (1) monocentric study; (2) reduced mechanical probe stability, handling difficulties with the double lumen bolt; (3) design changes to improve mechanical stability will require further study; (4) conflict of interest with Raumedic because of its support for the study. The conclusion drawn from our study is that the new multiparameter probe evaluated does measure ICP, TBr, and ptiO2. But all the initial data given in this paper have to be interpreted cautiously. A new study will be necessary when the mechanical stability of the new probe has been improved. PMID- 19295396 TI - Measurement of systolic pressure variation during graded volume loss using simple tools on Datex Ohmeda S/5 monitor. AB - Direct invasive arterial monitoring is performed routinely for all major neurosurgical procedures. Systolic pressure variation (SPV) used, independently or in combination with central venous pressure (CVP) allows optimal fluid management in hypovolemia and hemorrhage. This study aims to quantify SPV during graded hypovolemia using the simple technique described by Gouvea and Gouvea using Datex Ohmeda S/5, and to compare its reliability relative to other hemodynamic indicators of hypovolemia. Twenty anesthetized neurosurgical patients of ASA grade I and II patients were administered furosemide 0.5 mg/kg intravenously to obtain graded volume loss in the form of urine output. Invasive arterial pressure from radial artery and CVP were monitored using Datex OhmedaS/5 (Finland). Invasive arterial pressure label was changed to pulmonary artery label with the scale appropriate for arterial pressure. The trace was frozen in the wedge mode to reduce the sweep speed and the cursor was used to measure SPV and pulse pressure variation (PPV). Heart rate, systolic blood pressure, diastolic blood pressure, CVP at zero end-expiratory pressure, SPV and PPV are measured at baseline, and after a urine output of 200 and 500 mL. There was a significant correlation between volume loss and CVP, SPV, and PPV. The area under the curve of receiver operating characteristic analysis was >0.75 for CVP, SPV, and PPV. SPV of 7.5 mm Hg and a change of SPV by 4.5 mm Hg, a PPV of 4.5 and change in PPV by 2.5 mm Hg were the best cut-off values that corresponded to a volume change of 500 mL. This simple method enabled calculation of SPV without the computerized modules, and detected volume loss comparable to CVP. PMID- 19295397 TI - Median effective infusion dose (ED50) of alfentanil for monitored anesthesia care of percutaneous vertebroplasty of osteoporotic fractures. AB - Percutaneous vertebroplasty (PVP) consists of injecting small quantities of orthopedic cement to consolidate pathologic vertebral bodies. The procedure is brief but painful during vertebral puncture and cement injection requiring either general anesthesia or monitored anesthesia care with opioids. The optimal dose of alfentanil in this setting is unknown. Therefore, we sought to determine its median effective dose (ED50, or analgesic efficacy in 50% of patients) during PVP in none intubated, spontaneously breathing patients. After approval and informed consent of the Institutional Review Board, 50 patients (American Society of Anesthesiologists II-III, age 50 to 80, weight: 53 to 82 kg) with osteoporotic vertebral fractures were enrolled. The patients were premedicated with oral hydroxyzine 100 mg and had skin infiltration with 50 mg lidocaine before vertebral puncture. The prone position was adopted and oxygen was provided via a facemask. Noninvasive cardiorespiratory variables were monitored. Pain was evaluated by a numerical pain scale (NPS) where 0 represents no pain and 10 the worst tolerable pain. Alfentanil infusion was started 30 minutes before vertebral puncture. The initial dose was 2.0 mg/h. Thereafter, a 0.05 mg/h decrease or increase was applied to the next patient if analgesia was effective (NPS: 3 or less) or not (NPS>3) according to the Dixon method. The bootstrap resampling technique was used to calculate the ED50 and its 95% confidence limits. The latter was 1.05 mg/h (95% confidence interval, 1.0-1.2). Transient apnea (n=2) and nausea/vomiting (n=3) were observed. Given the median body weight of the patients (65 kg), we conclude that 0.27 microg kg/min of alfentanil provides effective analgesia for PVP under monitored anesthesia care. SUMMARY STATEMENT: PVP consists of injecting small quantities of orthopedic cement to consolidate pathologic vertebral bodies. The median effective dose (ED50) for pain relief during vertebral puncture and cement injection is 1.05 (95% confidence interval, 1.0-1.2) mg/h when infusion is started 30 minutes before the procedure. Given the median body weight of our study population (65 kg), this dose corresponds to 0.27 microg kg/min. PMID- 19295400 TI - Postoperative management of obstructive sleep apnea after transsphenoidal pituitary surgery. PMID- 19295401 TI - Determining the remifentanil dose-response relationship during craniotomies: the importance of intensity of the stimulus at the time of pharmacodynamic evaluation. PMID- 19295402 TI - The dilemma of anticoagulation for the heart, missing hemostasis for the brain, and the complications. PMID- 19295403 TI - Takotsubo syndrome after carotid endarterectomy. PMID- 19295404 TI - Report of the 36th annual meeting of society of neurosurgical anesthesia and critical care. PMID- 19295405 TI - Multilayered epithelium in mucosal biopsy specimens from the gastroesophageal junction region is a histologic marker of gastroesophageal reflux disease. AB - Barrett esophagus (BE) is defined as a columnar metaplasia of the distal esophagus that develops as a result of chronic gastroesophageal reflux disease (GERD). A distinctive type of multilayered epithelium (ME) that exhibits features of both squamous and columnar epithelium has been hypothesized to represent an early, or intermediate, phase in the development of BE. The aim of this prospective study was to evaluate the prevalence and specificity of ME in mucosal biopsies of the squamocolumnar junction (SCJ) from patients who had GERD, either with or without BE. During endoscopic examination of the esophagus, 2 biopsy specimens were obtained from across the SCJ from 27 patients with BE, 12 patients who had GERD without BE, and 14 controls who had no symptoms or endoscopic or histologic signs of GERD. ME was present at the SCJ in 33%, 33%, and 0% of BE, GERD, and control patients, respectively. Compared with control subjects, the prevalence of ME was significantly higher in both GERD and BE patients (P<0.05). In GERD patients without BE, ME was always detected adjacent to areas of cardia type mucosa composed of mucous glands. ME from GERD patients and BE patients had a similar immunophenotype, showing expression of the intestinal markers MUC2 and cdx-2 in 38% and 77% of cases, respectively. The prevalence of expression of these markers in ME was significantly different from nongoblet epithelium in control patients. Our results provide further evidence that ME may represent an early, transitional form of columnar metaplasia, and that ME may be used as a histologic marker of reflux disease in mucosal biopsies from the gastroesophageal junction region. PMID- 19295406 TI - SALL4 is a novel sensitive and specific marker of ovarian primitive germ cell tumors and is particularly useful in distinguishing yolk sac tumor from clear cell carcinoma. AB - Ovarian primitive germ cell tumors (GCTs) are uncommon tumors and sometimes pose diagnostic challenges. Among them, yolk sac tumor (YST) poses the greatest diagnostic difficulty and can be mistaken for clear cell carcinoma (CCC). Current immunohistochemical markers such as alpha-fetoprotein (AFP), glypican-3, cytokeratin (CK) 7, and epithelial membrane antigen (EMA) used to distinguish YST from CCC lack adequate sensitivity and specificity. Here by immunohistochemistry, we investigated a novel marker SALL4 in 98 GCTs (29 YSTs, 18 dysgerminomas, 6 gonadoblastomas, 6 embryonal carcinomas, 15 immature and 12 mature teratomas, 7 carcinoid tumors, 3 strumal carcinoids, and 2 struma ovarii) with particular interest of exploring SALL4 to distinguish YST from CCC. One hundred sixty-three non-GCTs including 45 CCCs were also stained. We found that SALL4 is strongly positive in more than 90% tumor cells in all YSTs, dysgerminomas, gonadoblastomas, and embryonal carcinomas. Variable SALL4 staining is seen in 11 of 15 immature teratomas. All other GCTs included in this study are negative for SALL4. Except 3 CCCs with focal SALL4 staining (<15% tumor cells), SALL4 is negative in the remaining 160 non-GCTs. We also compared SALL4 with AFP, glypican 3, CK7, and EMA in all YSTs and CCCs. AFP and glypican-3 are positive in 24 (83%) and 20 (69%) YSTs, respectively, whereas 16 (35%) and 13(28%) CCCs show positive AFP and glypican-3 staining, respectively. Three (10%) and 4 (14%) YSTs show focal (<2% tumor cells) CK7 and EMA staining, respectively. CK7 and EMA are positive in all 45 CCCs but 3 (7%) and 1 (2%) cases show staining in less than 30% tumor cells, respectively. Our findings indicate that SALL4 is a novel sensitive and specific marker for ovarian primitive GCTs. SALL4 is particularly useful in distinguishing YST from CCC and better than AFP, glypican-3, CK7, and EMA. PMID- 19295407 TI - Endocervical adenocarcinomas with prominent endometrial or endomyometrial involvement simulating primary endometrial carcinomas: utility of HPV DNA detection and immunohistochemical expression of p16 and hormone receptors to confirm the cervical origin of the corpus tumor. AB - Determining the primary site of a uterine adenocarcinoma can be problematic in hysterectomy specimens due to the overlapping morphology of endocervical adenocarcinomas and endometrial carcinomas, particularly when both the corpus (usually lower uterine segment) and endocervix are involved and precursor lesions are lacking or difficult to distinguish from intramucosal spread of carcinoma from one site to the other. Both preferential extension of endocervical adenocarcinomas into the endometrium (rather than deep cervical stroma) and myometrial invasion derived from the endometrial component are rarely encountered; to our knowledge, these unusual patterns of spread have not been detailed in prior reports. Clinicopathologic features of 10 endocervical adenocarcinomas (9 pure, 1 adenosquamous) with prominent endometrial or endomyometrial involvement were evaluated. Tumors were analyzed for the presence of human papillomavirus (HPV) DNA and by immunohistochemistry for expression of p16 and hormone receptors. Six cases had limited amounts of tumor in the cervix proper, with depths of invasion no greater than 5 mm in 4 and only adenocarcinoma in situ in 2. Four cases had cervical stromal invasion of more than 5 mm but all of these had greater amounts of horizontal extension into endometrium or endomyometrium. Four tumors extended into endometrium only and 6 had myoinvasion associated with the endometrial component. Five tumors were originally diagnosed as primary endometrial carcinoma with either cervical extension or concurrent endocervical adenocarcinoma in situ. HPV DNA was detected in both the cervical and corpus components in all tumors and all exhibited diffuse/strong p16 expression and decreased or absent expression of hormone receptors. These ancillary techniques are useful for clarifying the origin of uterine adenocarcinomas when morphologic features and tumor location are equivocal. These cases illustrate that dominant uterine corpus involvement (endometrial or endomyometrial) by primary endocervical adenocarcinoma can lead to misclassification as primary endometrial adenocarcinoma with cervical extension (Federation Internationale de Gynecologie et d'Obstetrique stage II), especially when endometrial extension of endocervical adenocarcinoma simulates complex atypical hyperplasia. A subset of misclassified endocervical adenocarcinomas may account for some HPV-positive uterine carcinomas reported as primary endometrial carcinomas. PMID- 19295408 TI - Pathologic features of ulcerative colitis in patients with primary sclerosing cholangitis: a case-control study. AB - BACKGROUND: The pathologic features of ulcerative colitis (UC) in patients with primary sclerosing cholangitis (PSC) are, essentially, unknown. One previous clinical study suggested that UC-PSC patients reveal a high rate of rectal sparing and backwash ileitis. The purpose of this study was to systematically evaluate the pathologic characteristics and distribution of colonic disease in UC PSC patients and to compare the results with a matched control group of UC patients without PSC. METHODS: Forty UC-PSC patients and 40 matched UC patients without PSC (controls) were identified from the files of 3 hospitals between the years 1989 and 2005. Clinical, endoscopic, and follow-up data (including incidence of pouchitis) were evaluated, and a detailed pathologic evaluation of biopsy and resection specimens (when available) was performed in a blinded fashion. The degree of activity and chronicity in mucosal biopsies and/or tissue from resection specimens was graded on a 5-point grading system (0 to 4), and each portion of the colon (cecum, ascending colon, transverse colon, descending colon, rectum) was assessed separately. Rectal sparing and patchiness of disease were evaluated, and scored as either absolute or relative depending on the complete absence of inflammatory disease in the former, or less inflammatory disease in the rectum compared with other parts of the colon in the latter. RESULTS: In this matched case-control study, UC-PSC patients presented at a significantly earlier age (24.5 y), had a higher prevalence rate of pancolitis (85%), and an overall significantly lower grade of inflammation in the colon (mean grade: 2.09+/-0.085) compared with UC controls (mean age: 33.8 y, pancolitis: 45%, inflammation grade: 2.59+/-0.92, P<0.05 for all comparisons). The incidence rate of absolute and relative rectal sparing (27.5%) and of patchy inflammatory disease proximal to the rectum (5.7%) was not significantly different between the UC-PSC cases and the UC controls (25% and 7.9%, respectively). UC-PSC patients had a higher prevalence rate of ileitis (35.7%) and pouchitis (42.8%), but the values were not significantly different from controls (26.9% and 26.6%, respectively). The incidence rate of dysplasia was similar between the 2 patient groups. CONCLUSIONS: UC patients with PSC show a propensity for more extensive, but less active, disease but are otherwise characterized by similar pathologic findings compared with UC patients without PSC. Rectal sparing and patchy disease activity is not characteristic of UC patients with PSC. PMID- 19295409 TI - Peripheral T-cell lymphomas with a follicular growth pattern are derived from follicular helper T cells (TFH) and may show overlapping features with angioimmunoblastic T-cell lymphomas. AB - Rare cases of peripheral T-cell lymphomas with follicular growth pattern (PTCL-F) have been recently reported, and their association with t(5;9)(q33;q22) involving ITK and SYK has been suggested. However, the clinicopathologic aspects of PTCL-F are poorly described and the normal cell counterpart of this subgroup of lymphoma is still unknown. Therefore, we analyzed the pathologic, phenotypic, and cytogenetic features of a series of 30 patients (range: 33 to 88 y) that showed histopathologic features of PTCL-F in at least 1 biopsy (n=30), either at initial presentation (n=26) or at relapse (n=4). Neoplastic cells were medium-sized clear cells that were CD4+ (24/27, 89%), CD10+ (21/29, 72%), BCL-6+ (14/19, 74%), and expressed programed death-1 (27/27, 100%), CXCL13 (23/27, 85%), and ICOS (11/11, 100%), markers of follicular helper T cells (TFH). Four of 22 patients (18%) had t(5;9)(q33;q22) detected by fluorescence in situ hybridization. Patients with clinical data available had multiple lymphadenopathies (25/28, 89%), stage III to IV diseases (17/26, 65%), B symptoms (7/27, 26%), and skin lesions (6/23, 26%). Three patients with sequential biopsies disclosed clinical and histopathologic features of angioimmunoblastic T-cell lymphoma at initial presentation. Our results show that this rare form of PTCL-F (1) has an immunophenotype indicative of derivation from TFH cells, (2) is associated with t(5;9) in a proportion of cases, and (3) shows some overlapping features with angioimmunoblastic T-cell lymphoma, raising the question of a possible relationship. PMID- 19295410 TI - Collagenous gastritis: histopathologic features and association with other gastrointestinal diseases. AB - Collagenous gastritis (CG) characterized by the deposition of a subepithelial collagen band and accompanying inflammatory infiltrate is a rare disorder. The natural history and pathogenesis of CG remain unclear. We describe the histologic features (23 gastric, 18 duodenal, and 4 colonic biopsies) and clinical findings of an additional 12 cases. Histologic features including active or chronic inflammation, surface epithelial injury, intraepithelial lymphocytosis, intestinal metaplasia, and Helicobacter pylori, and measurement of thickness of subepithelial collagenous band were evaluated in gastric biopsies. The clinical features, endoscopic findings, and follow-up were obtained and correlated with histologic features. There was an even number of males (n=6) and females (n=6). Four patients were children/young adults, 3 of whom (75%) presented with anemia and gastric nodularity. Eight patients were adults, 6 of whom (75%) had an associated autoimmune disease (1 with Hashimoto thyroiditis and polymyositis) or other intestinal disease (3 with celiac sprue, 1 with collagenous colitis, 1 with collagenous sprue), in contrast to none in the 4 children/young adults, P=0.06. The range of subepithelial collagen thickness was 15 to 120 microm in CG. The collagenous layer showed surface epithelial injury and entrapped inflammatory cells. On presentation, the thickened collagen distribution in the antrum and body was variably patchy and diffuse. Four (33%) patients showed lymphocytic gastritis (3 within the same biopsy); one of these patients also had celiac sprue and another had collagenous sprue. Three (25%) patients had celiac sprue (2 had duodenal biopsy proven and 1 had a clinical diagnosis of celiac sprue). An additional patient had duodenal biopsies showing collagenous sprue. Four patients had follow-up biopsies during a 3 to 119-month period after the diagnosis of CG. CG persisted on the follow-up gastric biopsies in 3 (75%) of the 4 patients, and the other patient had lymphocytic gastritis, a finding not seen in previous biopsies. CG is a rare disorder with a distinct presentation and association in pediatric and adult patients. An absence of associated intestinal and autoimmune diseases characterizes the pediatric population. Association with lymphocytic gastritis, celiac or collagenous sprue, collagenous colitis, and autoimmune disorders are frequently seen in adult patients. PMID- 19295411 TI - Polymorphisms of MMP-2 gene are associated with systolic heart failure risk in Han Chinese. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes responsible for protein degradation. MMP-2 has been demonstrated to play a pivotal role in myocardial remodeling process that occurs in congestive heart failure (HF). We hypothesized that MMP-2 genetic variations could be associated with systolic HF risk. METHODS: To test the association of single nucleotide polymorphisms of MMP-2 with systolic HF risk, we performed a hospital-based, case control study of 605 patients with systolic HF and 689 controls without HF. Three single nucleotide polymorphisms of MMP-2 (rs243864, rs243866, and rs17859821) were genotyped by restriction fragment length polymorphism methods. RESULTS: The genotype frequencies of MMP-2 rs243866 AA and AG in the control group were significantly higher than that in the case group (24.7% versus 17.9%, P < 0.01). Compared with the GG homozygotes, MMP-2 rs243866 A allele carriers had a significantly lower risk of systolic HF (adjusted OR 0.69, 95% CI 0.49-0.98; P = 0.035). Haplotype analysis indicated the haplotype GGG (rs243864-rs17859821 rs243866) was associated with higher risk of systolic HF (adjusted OR 2.05, 95% CI 1.08-3.89; P = 0.028). CONCLUSION: The findings of the current study suggest that MMP-2 rs243866 A allele was associated with lower risk of systolic HF in Han Chinese. PMID- 19295412 TI - Hemoglobin variability in chronic kidney disease: a cross-sectional study. AB - BACKGROUND: The ability to maintain dialysis patients' hemoglobin (Hgb) within narrow targets remains a significant clinical problem. This study was designed to determine the variability in Hgb values for patients with chronic kidney disease (CKD) receiving or not receiving erythropoiesis-stimulating agents (ESAs) compared with patients on dialysis receiving ESAs. METHODS: This cross-sectional review of anemia management in CKD and dialysis patients analyzed Hgb variability by patient-year, defined as the coefficient of variability calculated for individual patients. One hundred thirty-seven CKD patient-years and 350 dialysis patient-years were available for analysis. Hgb variability was defined as the coefficient of variability calculated as the individual patient's Hgb standard deviation divided by the patient's mean Hgb times 100. RESULTS: The coefficient of variability in Hgb values were significantly less in patients with CKD not treated with ESAs than in patients with CKD treated with ESAs whether they were receiving dialysis (medians: 3.96 versus 8.53%, P < 0.05) or not receiving dialysis (medians: 3.96 versus 7.37%, P < 0.05). CONCLUSION: CKD and hemodialysis patients receiving treatment with ESAs have significantly greater Hgb variability than patients with CKD not receiving ESAs. This finding suggests that the current practice pattern for the administration of exogenous ESAs is partly responsible for the observed Hgb variability. PMID- 19295413 TI - Experience of mycophenolate mofetil in 10 patients with autoimmune-related interstitial lung disease demonstrates promising effects. AB - BACKGROUND: Interstitial lung disease (ILD) is a frequent manifestation of connective tissue disease (CTD), especially systemic sclerosis (SSc), polymyositis-dermatomyositis, and rheumatoid arthritis. ILD related to CTDs heralds a poor prognosis and is associated with high mortality and 60% of patients have evidence of ILD. Cyclophosphamide (CYC) is extensively used in SSc ILD with moderate initial response but a poor long-term outcome, and is associated with significant toxicity. RESULTS: Mycophenolate mofetil (MMF) was administered to 10 patients with autoimmune-related ILD: 4 with SSc, 3 with rheumatoid arthritis, 2 with polymyositis, and 1 with systemic lupus erythematosus and Sjogren syndrome. Five patients received remote CYC infusion. Ten patients had improvement in alveolitis, symptoms (cough, dyspnea, and chest discomfort), perceived quality of life and activity levels. Four of 5 patients discontinued oxygen. Two of 8 repeat high-resolution computed tomography improved, 6 stabilized, none worsened. Pulmonary function testing in 1 of 9 patients showed worsening, 3 with improvement and 5 stabilized. Serial echocardiograms revealed no new pulmonary arterial hypertension and no worsening of preexisting pulmonary arterial hypertension. Very importantly, averaged prednisone dose decreased from 58 to 1.4 mg without worsening. CONCLUSION: MMF is safe, well tolerated, and allows reduction or discontinuation of prednisone without worsening of symptoms or objective progression of disease. MMF is less toxic and its targeted antifibrotic properties make it a potentially more effective agent than CYC that may supplant it as a first-line agent or provide sensible post-CYC maintenance or synergistic strategy in the treatment of CTD ILD. PMID- 19295414 TI - Rifampin hepatotoxicity associated with treatment of latent tuberculosis infection. AB - BACKGROUND: To determine the occurrence of hepatotoxicity associated with rifampin treatment of latent tuberculosis infection in patients from a public health tuberculosis clinic. METHODS: Evaluation of rifampin hepatotoxicity in adults aged >or=18 years from a database maintained from June 2001 to May 2007 in a public health department clinic. Rifampin 600 mg daily for 4 months was prescribed. Hepatotoxicity was defined as aspartate aminotransferase (AST) or alanine aminotransferase (ALT) levels more than 3 times the upper limit of normal (ULN) with symptoms or more than 5 times the ULN without symptoms. RESULTS: Rifampin therapy was initiated in 348 patients. Among 205 patients with evaluable data, 4 (1.95%, 95% confidence interval: 0%-4.33%) had AST or ALT levels >5 times the ULN (2 patients at 1 month and 2 patients at 3 months). Three of these patients had elevated AST/ALT at baseline; 1 had hepatitis C and 1 had an unconfirmed history of hepatitis. Adherence to clinic visits and prescribed treatment was poor. CONCLUSIONS: Rifampin hepatotoxicity associated with treatment of latent tuberculosis infection is rare. Our report suggests that hepatotoxicity is more likely in patients with baseline hepatic dysfunction and the need for increased vigilance in monitoring transaminases in these patients. PMID- 19295415 TI - Pancreas transplantation in African American patients using basiliximab induction. AB - The long-term outcome of pancreas transplant (PT) in African Americans (AA) using interleukin-2 receptor antibody induction has not been well documented. We retrospectively analyzed the 7-year outcomes of 45 AA and 73 white recipients of primary PT at our center. All PT were performed with enteric-systemic drainage. Basiliximab induction, tacrolimus, mycophenolic acid, and steroid maintenance were used as the primary immunotherapy. There was no difference by Kaplan-Meier analysis in patient (P = 0.94), pancreas graft (P = 0.76), or death-censored graft survival (P = 0.71) over 7 years between the AA and white groups. Clinically treated pancreas rejection episodes were slightly higher in AA than in white patients. Similarly, cytomegalovirus infection rates and comparable quality of graft function were noted in both groups over 7 years. Excellent long-term patient and pancreas graft survivals can be achieved in AA recipients of PT by using interleukin-2 receptor antibody induction and combination of tacrolimus, mycophenolic acid, and steroid maintenance. PMID- 19295416 TI - Radial augmentation index unmasks premature coronary artery disease in younger males. AB - OBJECTIVE: The augmentation index, a marker of arterial wave reflection, is considered to indicate cardiovascular risk burden, particularly in younger persons. We assessed whether the easily obtainable radial augmentation index (rAIx) is superior to systolic blood pressure (SBP) or pulse pressure (PP) in detecting atherosclerotic vascular disease at an early age. METHODS: We determined rAIx by applanation tonometry, SBP and PP in 152 male patients with or without invasively documented coronary artery disease (CAD). Ejection fraction (EF) was visually estimated by echocardiography or left ventricular angiography. RESULTS: In younger patients (age < or =60 years, EF > or =30%), rAIx was significantly higher in patients with CAD (79.8+/-13.5%, n=31) compared with patients without CAD (68.5+/-22.0%, n=21; P = 0.04), whereas SBP (121+/-16 vs. 131+/-18 mmHg, P=0.04) and PP (48.7+/-9.4 vs. 56.3+/-12.1 mmHg, P=0.01) were even lower in patients with CAD compared with patients without CAD. In patients aged < or =60 years, rAIx was highest when EF was less than 30% (90.0+/-9.2%) compared with patients with EF 30-54% (80.7+/-11.5%) or EF > or =55% (72.1+/-20.4%, P=0.01). In contrast, in patients above 60 years of age, rAIx, SBP or PP did not differ between patients with or without CAD and the rAIx tended to be lowest in patients with severely reduced EF (P=0.07). CONCLUSION: We conclude that premature vascular disease in younger patients with CAD is reflected only by an elevated rAIx, and not by SBP or PP. The rAIx is not suited to distinguish between older patients with high or low cardiovascular risk. Age seems to influence the interplay between rAIx and systolic heart function. PMID- 19295417 TI - Telephone intervention and quality of life in patients with breast cancer. AB - The aim of this quasi-experimental study was to examine the effectiveness of a telephone support intervention 1 week after surgery on the quality of life (QOL) of patients with breast cancer. The sample consisted of 228 patients with breast cancer allocated to an intervention group (n = 120) and control group (n = 108). The data were collected using Ferrans and Powers Quality of Life Index-Cancer Version (QLI-CV) and European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Breast Cancer Module (EORTC QLQ-BR23). The self reported QOL of patients with breast cancer was considered moderately high. Statistically significant associations were found between QOL and the demographic characteristics of age, education, and employment status and of having underage children. Statistically significant associations were found between QOL and clinical characteristics such as type of surgery and axilla treatment. The strongest predictors of poor QOL were age, control group, and type of surgery. Age was the strongest predictor of poor QOL in global QLI and in the health and functioning, socioeconomic, and family subscales. The patients' experiences show that the telephone intervention was helpful and the timing was appropriate. The QOL in patients with breast cancer was better in subscales of body image, future perspective, and postoperative side effects. The intervention group showed significantly better body image; they worried less about the future and had less postoperative side effects than the control group did. These results may help in discussing QOL issues and should be considered when planning and implementing interventions for patients with breast cancer. PMID- 19295418 TI - Early adulthood uprooted: transitoriness in young women with breast cancer. AB - Young women with breast cancer face a more aggressive disease and lower survival rate compared with those who are older, thereby confronting the potential finitude of life or transitoriness at a time when they are establishing their careers, developing partnerships, and building families. Little is known about the perspectives of young women with breast cancer in how they consider issues of their own mortality. Patients with stage I to III breast cancer aged 39 years and younger who were diagnosed within the last year (N = 16) were included in this qualitative study, which was a secondary analysis of texts written during an expressive writing intervention. Thematic analysis was done by 2 researchers, who reached consensus in coding and theme identification. Nine of the 16 participants (56%) wrote about their mortality (transitoriness). Three themes (being remembered, landscape of emotions and perspectives, and omnipresence of life's finitude) and 10 subthemes were identified from the rich narrative texts. Despite early-stage diagnosis and the beginning phases of adulthood, young women with breast cancer contemplate their mortality and its potential repercussions on their loved ones and their careers. Oncology professionals need to be sensitive to these concerns and provide opportunities for discussion and/or referrals for their young patients as they grapple with such potentially overwhelming issues. PMID- 19295419 TI - Psychometric testing of the Caregiver Quality of Life Index-Cancer on a Taiwanese family caregiver sample. AB - Family caregivers' quality of life (QOL) is an important indicator for end-of life care, but appropriate assessment tools are unavailable in Taiwan. This correlational study therefore tested the psychometric properties of the Caregiver Quality of Life Index-Cancer Mandarin version (CQOLC-M) in 359 dyads of cancer patients with terminal illness and their family caregivers from 5 teaching hospitals throughout Taiwan. Caregivers' QOL, spiritual well-being, social support, pain intensity, and economic situation were assessed using the CQOLC-M and 4 established scales. The CQOLC-M had internal consistency reliability of .87. Construct validity was supported by factor analysis, hypothesis testing, and known-group comparison. Exploratory factor analysis showed 7 underlying factors explaining 48.15% of the variance. Caregiver Quality of Life Index-Cancer Mandarin version scores correlated moderately with caregivers' social support (P < .01) and spiritual well-being (P < .01). Caregivers' QOL was inversely related to both patients' (F = 4.90; P = .008) and caregivers' average pain (t = -4.22; P < .001) in the past 24 hours. We suggest removing 4 items with factor loading of less than 0.4. Since caregivers' QOL depends on patients' and caregivers' physical dimensions, we recommend adding physical-related items to the CQOLC-M. Clinicians can improve caregivers' QOL not only by maintaining their health, social support, and spirituality but also by better managing the pain of patients with terminal illness. PMID- 19295420 TI - Establishing treatment fidelity in a coping and communication support telephone intervention for aging patients with advanced cancer and their family caregivers. AB - A randomized controlled trial of a nurse-delivered coping and communication support (CCS) intervention tailored to the preferences of middle-aged and older patients with advanced cancer and their family caregivers is tested for its value in improving outcomes for quality cancer care. An intervention study must be reliably delivered to estimate the accuracy of findings or make valid conclusions about the outcomes of research. The purpose of this article was to describe methodology for ensuring treatment fidelity in this patient-centered CCS intervention. Fidelity is maximized by ensuring that the CCS intervention is congruent with relevant theory, standardizing training and interventionist competence, and monitoring intervention delivery and documentation. We address unique challenges arising from using individualized interventions that preserve autonomy of the individual and are responsive to shifts in patients' needs and preferences over time. The challenge of nursing research on patient preference and coping communication requires both rigorous measurement of interventionist adherence to the intervention protocol and flexibility to allow for changing needs of patients and family caregivers. PMID- 19295421 TI - Identifying and responding to depression in adult cancer patients: evaluating the efficacy of a pilot communication skills training program for oncology nurses. AB - Depression is a common response among cancer patients to their diagnosis and treatment; however, it goes undetected by healthcare providers in about 50% of cases. Communication skills training has been suggested as means to help nurses detect and respond to patient depression. We developed and pilot tested a communication skills training workshop based around 6 strategies. The training program consisted of 2 half-day experiential workshops that included didactic teaching, exemplary video, and role play. The aim of the study was to evaluate the effectiveness of the communication skills training. Fifteen nurses were recruited from the ambulatory nursing service at (redacted). Standardized patient assessments were used to measure strategy uptake. The presence of each strategy was rated on a 4-point scale from "not attempted" to "successfully attempted." Nurses also completed evaluations of the training program. The nurses attempted 3 of 6 strategies more commonly after training, and a trend to significance was observed in a fourth strategy. The nurses reported more confidence to deal with patient depression and had greater self-efficacy. This short training program demonstrated success in improving nurse communication skills and confidence in dealing with patient depression. A larger trial of the training is planned. PMID- 19295422 TI - The effects of psychological status of the patients with digestive system cancers on prognosis of the disease. AB - To assess the association between psychological status and prognosis of patients with digestive system cancer, epidemiological investigations were conducted in 2 periods to obtain information about the patients' psychological status, survival time, and quality of life. Adjusted relative risks (95% confidence interval) of 1 year survival rate for anxiety and depression were 8.43 (1.18-60.20) and 9.62 (1.76-52.63), respectively. Relations between quality of life and depression and social support were statistically significant (P <.05). This study offers evidences that psychological status in a short-term postoperative period may affect 1-year survival rate, and poor psychological status and social support after being discharged from hospitals are associated with poor quality of life. PMID- 19295423 TI - Perceived intrafamily melanoma risk communication. AB - Melanoma is a skin cancer that can be deadly. Members of families with a strong history of melanoma have a high risk of melanoma occurrence or recurrence. Enhanced survival in these family members could be influenced by their knowledge of melanoma risk and by simple behaviors to decrease their risk or detect melanoma in its early, most curable, stage. Yet, there is minimal exploration on communication of risk or risk-modifying behaviors in melanoma at-risk families. In this study, we describe perceived intrafamily communication of melanoma risk. Using a qualitative descriptive approach, we examined in-depth interviews with 22 members of 8 families having 2 or more cases of melanoma. We identified 4 major themes: (1) awareness and understanding of risk, (2) families facilitate and hinder communication, (3) promoting melanoma prevention and detection in the family, and (4) an obligation to tell others. We discuss these findings in the context of extant knowledge of cancer risk communication in families at high risk for other cancers, impediments to cancer risk communication, remaining gaps in knowledge of this phenomenon, suggestions for hypothesis-driven research, and clinical implications that are applicable to these and other at-risk families. PMID- 19295424 TI - Development of a modified instrument to measure anticipatory grieving in Jordanian parents of children diagnosed with cancer: the Marwit and Meuser Caregiver Inventory Childhood Cancer. AB - The purpose of this article is to report on the development and field testing for validity and reliability of a modified version of the Marwit and Meuser Caregiver Inventory (MM-CGI) for the assessment of anticipatory grief among Jordanian parents of children with cancer (the MM-CGI Childhood Cancer). In 2006, a 50-item MM-CGI Childhood Cancer was administered to 140 Jordanian parents living with a child with cancer. The Cronbach alpha coefficient for the total instrument was .95, and Cronbach alpha coefficients for each of the 3 subscales was .91 for personal sacrifice burden, .90 for heartfelt sadness and longing, and .86 for worry and felt isolation. The construct validity of this instrument was supported by demonstrating a significant and positive correlation with the Anticipatory Grief Scale. The MM-CGI Childhood Cancer demonstrated strong convergent validity and excellent internal consistency reliability. However, further testing with a larger sample to facilitate factor analysis is needed to complete the validation process. PMID- 19295425 TI - The experience of spouses as informal caregivers for recipients of hematopoietic stem cell transplants. AB - Spouses often are the primary caregiver for the hematopoietic stem cell transplant (HSCT) recipients and participate in the transplant recipients' care throughout the HSCT trajectory. The purpose of this qualitative study was to explore and describe the experiences of spouses of HSCT recipients during the acute phase of the transplant trajectory. The participants were spousal caregivers (8 wives and 3 husbands) of transplant recipients. Each participant was interviewed 1 to 6 times. Data were analyzed using Spradley's domain analysis. As couples entered the transplant experience, spouses described a sense of dislocation from normal life. They were now riding a rollercoaster in the dark as they lived the uncertainty of this risky therapy. Spouses structured the uncertainty with rituals, formed a positive perspective, and envisioned the future. They described a caregiving role but also needed to balance "me and my world" with "us and our world." PMID- 19295426 TI - Health-promoting lifestyle factors of cancer survivors in Taiwan. AB - This study set out, using a prospective design, to examine the health-promoting lifestyle factors for cancer survivors. A questionnaire, based on the health promotion model, was used to collect self-reported data during clinical visits on the health-promoting lifestyle factors related to cancer and the health locus of control. A total of 57 cancer patients were followed up from the date of their interview to the date of their death from the disease. After controlling for age, sex, type of cancer, operation, radiotherapy, chemotherapy, and the number of months since cancer diagnosis, the external control and chance locus were identified as statistically significant preventive factors for these cancer patients. Although self-efficacy, social support, and health promotion lifestyle were all found to have no significant influences on the survival of these cancer patients, the survivors were found to be more likely to have higher scores on health promotion lifestyle factors and lower scores on self-efficacy. These results contribute to the information required for consideration of the cultural variables aimed at caring for cancer patients. PMID- 19295427 TI - Breast cancer survivorship program: testing for cross-cultural relevance. AB - Taking CHARGE, a theory-based self-management program, was developed to assist women with survivorship concerns that arise after breast cancer treatment. Few such programs have been evaluated for cultural relevance with diverse groups. This study determined the utility and cultural relevance of the program for African American (AA) breast cancer survivors. Two focus groups were held with AA women (n = 13), aged 41 to 72 years, who had completed primary treatment. Focus group participants assessed the program content, format, materials, and the self regulation process. Content analysis of audiotapes was conducted using an open, focused coding process to identify emergent themes regarding program relevance and topics requiring enhancement and/or further emphasis. Although findings indicated that the program's content was relevant to participants' experiences, AA women identified need for cultural enhancements in spirituality, self preservation, and positive valuations of body image. Content areas requiring more emphasis included persistent fatigue, competing demands, disclosure, anticipatory guidance, and age-specific concerns about body image/sexuality. Suggested improvements to program materials included portable observation logs, additional resources, more photographs of younger AA women, vivid colors, and images depicting strength. These findings provide the basis for program enhancements to increase the utility and cultural relevance of Taking CHARGE for AA survivors and underscore the importance of evaluating interventions for racially/ethnically diverse groups. PMID- 19295428 TI - Complementary and alternative medicine in pediatric allergic disorders. AB - PURPOSE OF REVIEW: Allergic disorders represent a serious public health problem in children. The chronic nature of these diseases and the fear of known side effects of synthetic drugs influence many families to seek complementary and alternative medicine. This review focuses on traditional Chinese medicine (TCM) herbal products and acupuncture for treating pediatric allergies. RECENT FINDINGS: Given the general safety profile and reputed efficacy, TCM are well received by the general population. However, compared with the long human history and popularity of the use of TCM, research into its efficacy and safety is still in its infancy. In the last 2-3 years, there have been more controlled studies of TCM for allergic asthma and allergic rhinitis. Several publications including ours indicate that some TCM herbal formulas are well tolerated and produce some level of efficacy. Some herbal formulas also showed beneficial immunomodualtory effects. Several preclinical studies demonstrated that the food allergy herbal formula-2 was effective in protecting against peanut anaphylaxis in animal models. Two TCM products have entered clinical trials in the United States for treating asthma and food allergy, respectively. Both of these trials include children. SUMMARY: Recent studies indicate that TCM therapy including herbal medicines and acupuncture for allergic disorders in children is well tolerated. There are also promising clinical and objective improvements. More controlled clinical studies are encouraged. PMID- 19295429 TI - Gene-environmental interaction in the development of atopic asthma: new developments. AB - PURPOSE OF REVIEW: Over recent years, major advances have occurred in understanding of the role of atopy in asthma. To accommodate these advances requires a revision in the paradigm that asthma is an atopic disease. In turn, a revision in the focus of gene-environment studies is now needed. The aim of this review, therefore, is to provide a new perspective to the topic of the role of genes and environment in the development of symptoms in atopic asthma. RECENT FINDINGS: Recent data from prospective birth-cohort studies has shown that atopic events appear to play a limited role in the development of asthma and even less of a role in significant acute wheeze in children. Overwhelmingly, acute respiratory viral infection, not acute allergen exposure, is the major environmental cause of acute wheezing episodes in asthmatic children. This knowledge has led to the examination of the genetics of the immune system, particularly genes controlling innate immune responses, with respect to viral defences - an important area of gene-environment interaction in asthma. SUMMARY: Advances in knowledge of the genetics of viral defence have contributed to improved insight into asthma in children and could lead to effective new antiviral asthma therapies. PMID- 19295430 TI - The role of recombinant-activated factor VII in bleeding trauma patients. AB - PURPOSE OF REVIEW: Exsanguinating hemorrhage and postshock organ failure account for 35-40% of deaths from trauma, and there is an increasing recognition of the importance of coagulopathy in the evolution of this disease. RECENT FINDINGS: Since 1999, case reports, small series, retrospective studies and a few controlled trials have reported the use of recombinant-activated factor VII (rFVIIa) as an adjunct for reversal of coagulopathy in trauma patients, and numerous other publications have examined the use of rFVIIa in related conditions such as traumatic brain injury, hemorrhagic stroke and uncontrolled surgical bleeding. SUMMARY: We present a brief discussion of the mechanism of action of rFVIIa and its role in facilitating hemostasis and a review of the recent medical literature on the use of rFVIIa in trauma patients, including current guidelines and controversies. PMID- 19295431 TI - Can efficient supply management in the operating room save millions? AB - PURPOSE OF REVIEW: Supply expenses occupy an ever-increasing portion of the expense budget in today's increasingly technologically complex operating rooms. Yet, little has been studied and published in the anesthesia literature. This review attempts to bring the topic of supply management to anesthesiologists, who play a significant role in operating room management. RECENT FINDINGS: Little investigative work has been performed on supply management. Anecdotal reports suggest the benefits of a perpetual inventory system over a periodic inventory system. A perpetual inventory system uses utilization data to update inventory on hand continually and this information is linked to purchasing and restocking, whereas a periodic inventory system counts inventory at some regular intervals (such as annually) and uses average utilization to set par levels. SUMMARY: On the basis of application of operational management concepts, ways of taking advantage of a perpetual inventory system to achieve savings in supply expenses are outlined. These include linking the operating room scheduling and supply order system, distributor-driven just-in-time delivery of case carts, continual updating of preference lists based on utilization patterns, increasing inventory turnovers, standardizing surgical practices, and vendor consignment of high unit cost items such as implants. In addition, Lean principles of visual management and elimination of eight wastes may be applicable to supply management. PMID- 19295432 TI - Acute pain in children and adults with sickle cell disease: management in the absence of evidence-based guidelines. AB - PURPOSE OF REVIEW: Acute, vaso-occlusive pain is the most characteristic complication of sickle cell disease (SCD). Although there has been rigorous work examining the pathogenesis of vaso-occlusion, fewer studies have focused on approaches to the clinical management of acute pain. In this review, we will examine the epidemiology and management strategies of acute pain events and we will identify limitations in the best available studies. RECENT FINDINGS: Most acute pain events in adults with SCD are managed at home without physician contact. Prior descriptions of the natural history of pain episodes from the Cooperative Study of Sickle Cell Disease relied on physician contact, limiting the generalizability of these findings to current practice. Patient-controlled analgesia has replaced on-demand therapy to become the standard for management of severe pain events in children and adults with SCD requiring hospital admission. SUMMARY: Unfortunately, most clinical practice guidelines for the management of acute pain are not based on randomized clinical trials. As a result, our practice of pain management is primarily limited to expert opinion and inferences from observational studies. Additional clinical trials in management of acute pain in children and adults with SCD are critical for the development of evidence-based guidelines. PMID- 19295433 TI - Pediatric genitourinary tumors. AB - PURPOSE OF REVIEW: We will review the 2007/2008 literature on pediatric genitourinary tumors. RECENT FINDINGS: Newly identified constitutional epigenetic defects in Wilms tumor genes extend the understanding of Wilms tumor risk in children lacking syndromic features, and add to the complexity of the pathogenesis of these tumor suppressor genes. Pediatric renal cell carcinoma has distinct molecular characteristics and clinical associations from the adult counterpart. The pathway from PAX3-FKHR translocation to the development of rhabdomyosarcoma tumors has been further elucidated. SUMMARY: Therapeutic strategies continue to be driven by developments in molecular diagnostics in pediatric genitourinary tumors. PMID- 19295434 TI - Outcomes and follow-up strategies for patients on active surveillance. AB - PURPOSE OF REVIEW: Prostate cancer is now the most commonly diagnosed solid tumor in American men. Autopsy studies show the uniquely high prevalence rates of small, indolent tumors in men dying of other causes. These findings have led to an increased interest in managing men with prostate cancer and low risk features expectantly, with close observation for early signs of progression. This approach allows one to limit prostate cancer treatment and any risk of related morbidity to the men who will benefit the most from active intervention. RECENT FINDINGS: Several centers have published their results with active surveillance and delayed selective therapy for men with low grade, early prostate cancer. Although median follow up from these studies is relatively short, the outcomes appear favorable. About one third of men will receive treatment after 3-5 years of surveillance and quality of life remains high. SUMMARY: Data from these reports provide patients and clinicians with the early experiences and expectations with active surveillance for prostate cancer. They also provide a framework for selecting men for this approach and for following them over time. Prospective, randomized trials comparing active surveillance with standard interventions are underway. PMID- 19295435 TI - Summer temperature-related mortality: effect modification by previous winter mortality. AB - BACKGROUND: Several studies have described seasonal patterns of mortality, with rates higher in winter and lower in summer. Few researchers, however, have analyzed how the mortality rate in winter may influence the temperature-mortality association in the following summer. In the present paper, we addressed the question of whether the association between summer temperature and mortality among the elderly is modified by the previous winter mortality rate. METHODS: We selected all deaths in Rome during 1987-2005 among persons 65 years old or older. We collected data on daily mean temperature and humidity. We estimated the effect of summer apparent temperature on mortality by using a time-series approach, and tested the effect modification based on the mortality rate during the preceding winter. RESULTS: The effect of summer apparent temperature on mortality was stronger in years characterized by low mortality in the previous winter (relative risk for days at 30 degrees C vs. days at 20 degrees C = 1.73 [95% confidence interval = 1.50-2.01]), as contrasted with years with medium (1.32 [1.25-1.41]) or high winter mortality (1.34 [1.17-1.55]). The percentages of attributable risks for summer heat were 28%, 18%, and 18% for years characterized by low, medium, or high winter mortality rates respectively. CONCLUSIONS: Low-mortality winters may inflate the pool of the elderly susceptible population at risk for dying from high temperature the following summer. PMID- 19295436 TI - Rejoinder: Progress in understanding the relationship between beryllium exposure and lung cancer. PMID- 19295437 TI - Targeted BCG vaccination against severe tuberculosis in low-prevalence settings: epidemiologic and economic assessment. AB - BACKGROUND: BCG vaccine protects against the severe forms of tuberculosis (TB) in children. Several low-prevalence countries are reviewing their policy, usually shifting from universal vaccination to vaccination of infants in high-risk groups only. We combined an epidemiologic analysis with a cost-effectiveness analysis to evaluate the cost-effectiveness of targeted strategies. METHODS: We fitted a static model to the data to estimate vaccine efficacy and risk of disease. We applied our method to the Dutch situation, analyzing severe TB cases in high-risk group children age 0-5, between 1996 and 2003. We considered the current strategy targeting immigrant children from high-incidence countries, and a proposed strategy additionally targeting children from 3 lower-incidence, but higher immigration, countries. RESULTS: In the absence of vaccination, the annual risk of developing severe TB for a child in the current target group is 3/100,000, while BCG vaccination reduces this risk by 73%. Therefore about 9000 children would need to be vaccinated to prevent 1 case. Vaccinating children from high incidence countries would then cost about Euro 4,500 per discounted disability adjusted life year averted. In the extended target group, the risk of disease is somewhat lower with a similar vaccine effectiveness, so costs are raised. CONCLUSIONS: The current Dutch BCG strategy, as well as the proposed inclusion of immigrant children from Turkey, Surinam and former Yugoslavia, is on average cost effective. However, the low number of both vaccinated and unvaccinated severe TB cases leads to broad confidence intervals on vaccine efficacy, highlighting the difficulty associated with decision-making in low-prevalence settings. PMID- 19295438 TI - Does fentanyl really need to be omitted in favour of sevoflurane in day care surgery? PMID- 19295439 TI - Are newly diagnosed columnar-lined oesophagus patients getting younger? AB - OBJECTIVES: The prevalence of columnar-lined oesophagus seems to have increased steadily in the past three decades in Europe and North America. Although the vast majority of columnar-lined oesophagus will not progress to malignancy, it is nevertheless important to identify the risk factors associated with this condition. This study investigates whether there has been a change, at diagnosis, in age of columnar-lined oesophagus patients between 1990 and 2005, or an increase in the number of patients aged less than 50 years. METHODS: Data on age of diagnosis were abstracted from medical records of 7220 patients from 19 centres registered with UK National Barrett's Oesophagus Registry, between the years 1990 and 2005. Linear regression analysis was carried out to assess any trends in the mean age of diagnosis. RESULTS: Overall there was a mean decrease in age at diagnosis for each 1-year increase in time. This equated to a mean decrease of 3 years over the study period, 1990-2005 with the greatest difference being seen in female patients. About 18% of patients in the study were aged less than 50 years at the time of diagnosis. With this group also, the trend was similar, with an increase in the number of patients aged less than 50 years, at the time of diagnosis, with increasing years. CONCLUSION: The mean age of diagnosis of columnar-lined oesophagus has decreased between the years 1990 and 2005 in both men and women, more so in women. This is also reflected in an increase in newly diagnosed columnar-lined oesophagus patients below the age of 50 years. PMID- 19295440 TI - Association of gastric disease with polymorphisms in the inflammatory-related genes IL-1B, IL-1RN, IL-10, TNF and TLR4. AB - OBJECTIVES: This study aimed to investigate if single nucleotide polymorphisms (SNPs) in a series of inflammatory genes were associated with the development of the most common pathologies thought to precede gastric cancer development namely; Helicobacter pylori (H. pylori)-associated gastritis and intestinal metaplasia. METHODS: A total of 250 patients were genotyped for 11 SNPs in the IL-1B, IL-1RN, TNF, TLR4 and IL-10 genes. The study population comprised H. pylori uninfected ('normal') control patients (n=96), H. pylori-positive gastritis (n=91) and intestinal metaplasia patients (n=63). Genotyping was performed using Taqman allelic discrimination assays. Odds ratios for gastric disease groups were adjusted for potential confounding factors. RESULTS: No differences were identified in frequency of carriage, or homozygosity, for any of the 'risk' alleles investigated across the patient groups. No evidence was found to suggest an association with increased risk of developing either chronic gastritis or intestinal metaplasia with SNPs in the IL-1B, IL-1RN, TNF, TLR4 and IL-10 genes or haplotypes tested. CONCLUSION: This study found no evidence of an association with increased risk of developing either chronic gastritis or intestinal metaplasia with the SNPs or haplotypes tested. PMID- 19295441 TI - Metformin prevents myocardial reperfusion injury by activating the adenosine receptor. AB - Metformin improves cardiovascular outcomes in patients with type 2 diabetes compared with other glucose-lowering drugs. Experimental studies have shown that metformin can increase the intracellular concentration of adenosine monophosphate, which is a major determinant of the intracellular formation of adenosine. We hypothesize that metformin, given at reperfusion, can limit myocardial infarct size due to increased adenosine receptor stimulation. Isolated perfused hearts from Sprague-Dawley rats were subjected to 35 minutes of regional ischemia and 120 minutes of reperfusion. Perfusion with metformin (50 microM) for the first 15 minutes of reperfusion reduced infarct size (percent area at risk) from 42% +/- 2% to 19% +/- 4% (n >or= 6; P < 0.01), which was blocked by a concomitant perfusion with the adenosine receptor antagonist 8-p sulfophenyltheophylline (100 microM; 43% +/- 3%) or nitrobenzylthioinosine (a blocker of transmembranous adenosine transport; 1 microM; 45% +/- 5%). In addition, intravenous administration of metformin (5 mg/kg) reduced infarct size in a rat in situ model of myocardial infarction (34% +/- 6% vs. 62% +/- 5%; P < 0.01), which was completely abolished by 8-p-sulfophenyltheophylline (61% +/- 3%). We conclude that metformin, given at reperfusion, reduces infarct size in a rat model of myocardial infarction, which is critically dependent on adenosine receptor stimulation, probably via increased intracellular formation of adenosine. PMID- 19295442 TI - Antiproliferative action of cudraflavone B, isolated from Cudrania tricuspidata, through the downregulation of pRb phosphorylation in aortic smooth muscle cell proliferation signaling. AB - Cudrania tricuspidata has been proposed to possess anti-inflammatory, antioxidant, hepatoprotective, and antitumor activities. Although cudraflavone B, isolated from the root bark of C. tricuspidata, has a variety of pharmacological effects, its effects on rat aortic smooth muscle cells (RASMCs) are unclear. In the present study, cudraflavone B was found to inhibit cell proliferation and DNA synthesis in cultured RASMCs. Pretreatment with cudraflavone B (0.1-4 microM) suppressed platelet-derived growth factor-BB (PDGF-BB)-stimulated cell number in a concentration-dependent manner. The inhibition percentages were 19.7%, 36.4%, 52.3%, and 99.1% at concentrations of 0.1, 1, 2, and 4 microM, respectively. Moreover, cudraflavone B inhibited [H]-thymidine incorporation into DNA in RASMCs in response to 25 ng/mL PDGF-BB. PDGF-BB-stimulated DNA synthesis was significantly reduced by 15.9%, 31.7%, 43.1%, and 78.2% at concentrations of 0.1, 1, 2, and 4 muM, respectively. Thus, cudraflavone B blocked the PDGF-BB-inducible progression through G0/G1 to S phase of the cell cycle in synchronized cells. Furthermore, PDGF-BB-induced phosphorylation of retinoblastoma protein (pRb), the hyperphosphorylation of which is a hallmark of the G1-S transition in the cell cycle, was significantly inhibited by cudraflavone B. Because pRb phosphorylation is regulated by cyclin-dependent kinases (CDKs), we investigated the expression of CDK2, CDK4, cyclin E, and cyclin D1 and the CDK inhibitors p21 and p27. Treatment with cudraflavone B downregulated the cyclins and CDKs and upregulated the expression of p21 and p27, a CDK inhibitor. These findings suggest that cudraflavone B inhibits RASMC proliferation via the induction of p21 and p27 expression and subsequent cell cycle arrest with reduction of pRb phosphorylation at the G1-S phase. PMID- 19295443 TI - Characterizing the mechanisms of insulin vasodilatation of normal and streptozotocin-induced diabetic rat aorta. AB - The mechanism by which insulin causes vasodilatation remains unclear, so we explored this in aortic rings from normal Wistar Kyoto and streptozotocin-induced diabetic rats. Insulin-induced relaxation of phenylephrine-contracted [endothelium (ED) intact or denuded] aortic rings was recorded in the presence or absence of various drug probes. Insulin relaxant effect was more in ED-intact than in-denuded tissues from normal or diabetic rats. l-NAME or methylene blue partially inhibited insulin effect in ED-intact but not the ED-denuded tissues, whereas indomethacin (cyclooxygenase inhibitor) had no effect on any of the tissues, indicating that insulin induces relaxation by ED-dependent and independent mechanisms, the former via the NOS-cyclic guanosine monophosphate but not the cyclooxygenase pathway. The voltage-dependent K channel (KV) blocker (4 aminopyridine) inhibited insulin action in all the tissues (normal or diabetic, with or without ED), whereas the selective BKCa blocker, tetraethylammonium, inhibited it in normal (ED intact or denuded) but not in diabetic tissues, indicating that KV mediates insulin action in normal and diabetic tissues, whereas the BKCa mediates it only in normal tissues, with possible pathophysiologic absence in diabetic tissues. The inward rectifier K channel (Kir) blocker (barium chloride) significantly inhibited insulin effect only in ED intact or -denuded diabetic tissues, whereas the KATP channel blocker, glibenclamide, inhibited it only in the ED-denuded diabetic tissues, suggesting that Kir channels mediate insulin-induced relaxation in ED-intact or -denuded diabetic tissues, whereas the KATP channel mediates it in ED-denuded diabetic tissues. All the agents combined did not abolish insulin action, suggestive of a direct vasodilatory effect. In conclusion, insulin causes vasodilatation in normal and diabetic tissues via ED-dependent and -independent mechanisms differentially modulated by K channels, some of which functions are altered in diabetes and thus are potential therapeutic targets. PMID- 19295444 TI - Chronotropic effects of azelnidipine, a slow- and long-acting dihydropyridine type calcium channel blocker, in anesthetized dogs: a comparison with amlodipine. AB - Azelnidipine, a dihydropyridine calcium channel blocker unlike other dihydropyridine calcium channel blockers, does not increase but slightly decreases heart rate (HR) in clinical settings. In the present study, the mechanism for the HR-lowering action characteristic of azelnidipine was investigated in anesthetized dogs. In the in situ perfused sinus node preparation, the negative chronotropic action of azelnidipine was almost 5 times more potent than that of amlodipine. When injected intravenously in intact anesthetized dogs, both drugs produced a long-lasting blood pressure reduction, but the action of azelnidipine developed more slowly than that of amlodipine. Azelnidipine hardly affected HR at lower doses and decreased HR at higher doses. On the other hand, amlodipine consistently induced slight tachycardia. The HR lowering effect of azelnidipine in autonomically blocked dogs was not much different from that in intact dogs, whereas tachycardia induced by amlodipine was eliminated in autonomically blocked dogs. However, neither drug affected the HR gain of baroreceptor reflex induced by changing carotid sinus pressure. These results suggest that compared with amlodipine, azelnidipine has a greater intrinsic negative chronotropic action and induces a lower level of reflex tachycardia most probably due to a slower development of blood pressure-lowering effects and that these characteristics of azelnidipine underlies its HR-lowering effects when administered systemically. PMID- 19295445 TI - Combining sirolimus-eluting stents and external irradiation in cholesterol-fed rabbits increased incomplete stent apposition and decreased re endothelialization. AB - Restenosis after the implantation of a drug-eluting stent or after vascular irradiation therapy shares similar physiopathological mechanisms. No experimental data are currently available on vascular wall behavior after external irradiation on arteries stented with sirolimus-eluting stents (SES). Ten New Zealand white rabbits received a 0.5% cholesterol-enriched chow for 1 month. Bilateral iliac artery stent implantation was then performed with an SES (Cypher; Cordis Corp). The animals were randomized into either an irradiated group (I, 2 Gy external x ray irradiation, n = 5) or a control group (C, n = 5). The cholesterol-enriched chow was continued for 1 additional month after stent implantation. The stented arteries were harvested for histological analyses. The number and the percentage of incompletely apposed stents struts (IASS) were significantly higher in irradiated versus control group (3.05 +/- 0.46 vs. 1.57 +/- 0.27 IASS, P < 0.01, and 28.44% +/- 3.97% vs. 15.2% +/- 2.46% of IASS, P < 0.01, respectively). The mean neointimal thickness behind the IASS was also higher in the irradiated group (I: 28.3 +/- 2.5 microm vs. C: 18.2 +/- 2.3 microm, P < 0.01). Re endothelialization was lower in irradiated group (I: 44.6% +/- 17.5% vs. C: 75.2% +/- 5.7%, P < 0.01). The present study revealed that low-dose external irradiation increased incomplete stent apposition and reduced re endothelialization of SES. These results underscore the potential deleterious cumulative side effects of these 2 procedures to prevent restenosis. PMID- 19295446 TI - Investigation of autonomic function and orocecal transit time in patients with nonalcoholic cirrhosis and the potential influence of these factors on disease outcome. AB - BACKGROUND: The presence of autonomic dysfunction in nonalcoholic cirrhosis and its influence on intestinal transit and disease outcome still need clarification. GOALS: To investigate the function of the autonomic nervous system in patients with nonalcoholic cirrhosis and the possible associations among autonomic dysfunction, severity of liver disease, disturbed intestinal transit, and the development of complications during follow-up. STUDY: Measurements of heart rate variability obtained by analysis of 24-hour ambulatory electrocardiographic recordings to assess autonomic function and lactulose breath hydrogen test to determine orocecal transit time were performed in 32 patients with nonalcoholic cirrhosis divided into Child A and B. RESULTS: Child B patients showed significantly lower values (P<0.05) of those parameters reflecting parasympathetic (high frequency, log-transformed high frequency, pNN50) and sympathetic function (low frequency, log-transformed low frequency) in comparison with controls and Child A patients. Orocecal transit time values were significantly (P=0.02) higher in Child B patients than in controls, but no relationship was found between delayed orocecal transit time and autonomic dysfunction. During follow-up, 42% of Child B patients developed encephalopathy. This complication was significantly associated with autonomic dysfunction. In addition, in the 4 patients who died the parameters reflecting parasympathetic function were significantly reduced in comparison with those of survivors. CONCLUSIONS: Autonomic dysfunction and delayed intestinal transit are related to the severity of disease in nonalcoholic cirrhosis. Autonomic dysfunction seems to predispose cirrhotic patients to the development of encephalopathy and may be associated with a poor prognosis of these patients. PMID- 19295447 TI - The clinical utility of esophageal manometry. AB - BACKGROUND AND AIMS: Esophageal manometry (EM) is performed to evaluate symptoms of chest pain and dysphagia, although its clinical utility is not known. The aim of this study was to evaluate the clinical utility of EM by determining whether EM provides new information, changes diagnoses, or alters patient management. METHODS: Before performing EM, referring providers noted indications for the test, symptoms, previous tests performed, and medication use. After EM was completed, a follow-up questionnaire ascertained whether EM provided new information or changed the patient's diagnosis or management plan. Patients provided demographic information. RESULTS: During a 12-month period, 569 EMs were performed and 444 were available for inclusion; 286 fully completed questionnaires were returned (64%) and are the basis for this analysis. The mean age (+/-SD) at the time of manometry was 52 (+/-15) years; 58% were women; 98% were white. EM was requested to assist placement of a pH measuring device (34%), and to evaluate symptoms of dysphagia (29%), chest pain (12%), or acid reflux (11%). Overall, 64% of EM were abnormal; 81% in gastroesophageal reflux disease patients, 74% in dysphagia, and 59% in chest pain. New information was obtained in 87% of patients, whereas a change in diagnosis occurred in 30% of patients, and management changed in 44% of patients. CONCLUSIONS: EM is a clinically useful test because it frequently provides new information and often changes patient diagnosis and management. The clinical utility of EM is greatest in patients with dysphagia. PMID- 19295448 TI - Treatment outcomes with pegylated interferon and ribavirin for male prisoners with chronic hepatitis C. AB - GOALS: To report our experience with pegylated interferon and ribavirin treatment of hepatitis C virus (HCV) RNA-positive inmates at the Rhode Island Department of Corrections. BACKGROUND: An estimated 1 out of 3 HCV-infected individuals will spend time in a jail or prison within a 1-year period, making prisons a unique setting for management of chronic HCV. STUDY: Chart review of all inmates identified as having initiated HCV treatment between October 2000 and April 2004. HCV-infected individuals were identified by HCV antibody screening at intake for known risk factors, elevated aminotransferase levels, or per individual request. Treatment followed standard guidelines with weight-based dosing of pegylated interferon-alpha2b and ribavirin. End points were completion of therapy plus 6 months for sustained virologic response (SVR), therapy discontinuation, and loss to follow-up. RESULTS: The cohort included 71 male patients, was mostly white (80%), and genotype 1 (65%). All 9 African Americans (AA) had genotype 1. Of 59 patients having liver biopsy, 41 had early stage disease. Overall SVR was 28%. Response rate was lower for genotype 1 compared with genotypes 2 and 3 (SVR 18% vs. 60% and 50%). Of inmates with genotype 1, no difference existed in treatment response by race (SVR 22% AA vs. 18% white). Thirty-three patients completed treatment, 26 stopped for side effects, and 5 for initial nonresponse. Eleven were lost to follow-up. CONCLUSIONS: Acceptable HCV treatment outcomes can be achieved in prisons. Our small study indicates no difference in treatment response by AA versus white race for genotype 1. PMID- 19295449 TI - Newer antidepressants and gabapentin for hot flashes: a discussion of trial duration. AB - OBJECTIVE: Information regarding the ideal length of hot flash trials is scarce. In the literature, hot flash trial durations have commonly varied from 4 to 12 weeks. This article is devoted to providing scientific data to better ascertain how long it is necessary to conduct hot flash trials with newer centrally acting agents. METHODS: Individual participant data were collected from all known published, through December 2007, randomized, placebo-controlled, double-blinded clinical trials regarding the use of newer antidepressants and gabapentin for hot flash relief. Trials that studied periods longer than 4 weeks were included for this project. Profile analysis was applied to the hot flash activity longitudinal data for each study individually, allowing a comparison of data collected for 6 to 12 treatment weeks versus data collected for only 4 treatment weeks. RESULTS: Ten studies were identified, five of them fulfilled the eligibility criteria for this investigation, three evaluating gabapentin, and two newer antidepressants. Flatness tests from a profile analysis did not provide any evidence that hot flash activity increased or decreased between week 4 and time periods up to 12 weeks. CONCLUSIONS: Changes in hot flash scores from newer antidepressants and gabapentin are apparent within 4 weeks of therapy. Available data indicate that hot flash treatment efficacy, compared with that of placebo, remains stable for up to 12 weeks of follow-up. PMID- 19295450 TI - Brain-region responsiveness to DT56a (Femarelle) administration on allopregnanolone and opioid content in ovariectomized rats. AB - OBJECTIVE: The natural selective estrogen receptor modulator DT56a (Femarelle), derived from soybean, has been shown to relieve menopausal vasomotor symptoms with no effect on sex steroid hormone levels or endometrial thickness.The purpose of the present study was to evaluate the neuroendocrine effect of DT56a administration through the evaluation of brain content of allopregnanolone (AP), an endogenous neurosteroid gamma-aminobutyric acid agonist with anxiolytic properties, and through the assessment of beta-endorphin (beta-END), the endogenous opioid implicated in pain mechanism, emotional state, and autonomic control. METHODS: Five groups of Wistar ovariectomized (OVX) rats received one of the following treatments: oral DT56a administration at doses of 6, 12, 60, and 120 mg kg(-1) day(-1) or estradiol valerate (E2V) at a dose of 0.05 mg kg(-1) day(-1) for 14 days. One group of fertile and one group of OVX rats receiving placebo were used as controls. The concentration of AP was assessed in the frontal and parietal cortex, hippocampus, hypothalamus, anterior pituitary, and serum, whereas the content of beta-END was evaluated in the frontal and parietal cortex, hippocampus, hypothalamus, neurointermediate lobe, anterior pituitary, and plasma. RESULTS: DT56a increased AP levels in all brain areas analyzed and in serum, with a classical dose-related curve in comparison with OVX rats. In some brain areas, such as the frontal cortex, the parietal cortex, and the anterior pituitary, positive results were found even with the administration of a lower DT56a dose of 60 mg kg(-1) day(-1), attaining AP levels in the range of those in animals treated with E2V. Similarly, beta-END levels were enhanced in selected brain areas such as the hippocampus, the hypothalamus, the neurointermediate lobe, and the anterior pituitary in comparison with those in OVX rats, in which the increase of the opioid was dose related and in the range of those in rats treated with E2V. CONCLUSIONS: This study demonstrated that DT56a positively affects brain neurosteroidogenesis and the opiatergic system: DT56a exerts an estrogen-like effect on selective areas related to mood, cognition, and homeostasis control, presenting a specific pattern of interaction with the brain function. These findings may, in part, explain the clinical effect of DT56a on menopausal symptoms. PMID- 19295451 TI - Dopamine D1-like receptor activation induces brain-derived neurotrophic factor protein expression. AB - Recent studies showed that dopamine or D1 receptor-selective agonists increased brain-derived neurotrophic factor (BDNF) mRNA and protein expression in neuronal cultures, and this action was blocked by SCH23390. Moreover, SKF38393 activated Trk receptors and downstream signaling in striatal neurons. This study examined whether dopamine agonists induce the expression of BDNF protein in rat brain tissue. Acute slice preparations were incubated with dopamine agonists in Hibernate A medium and BDNF protein was measured by a sensitive enzyme-linked immunosorbent assay. Results showed that dopamine increased BDNF in tissue slices after 24 h of incubation. Furthermore, SKF38393 produced a significant increase in BDNF protein in striatal and hippocampal tissue slices. These findings suggest that the induction of BDNF expression may constitute a downstream response to D1 like dopamine receptor activation. PMID- 19295452 TI - Short-term modulation of extracellular signal-regulated kinase 1/2 and stress activated protein kinase/c-Jun NH2-terminal kinase in pancreatic islets by glucose and palmitate: possible involvement of ceramide. AB - OBJECTIVES: The effect of glucose and palmitate on the phosphorylation of proteins associated with cell growth and survival (extracellular signal-regulated kinase 1/2 [ERK1/2] and stress-activated protein kinase/c-Jun NH2-terminal kinase [SAPK/JNK]) and on the expression of immediate early genes was investigated. METHODS: Groups of freshly isolated rat pancreatic islets were incubated in 10 mmol/L glucose with palmitate, LY294002, or fumonisin B1 for the measurement of the phosphorylation and the content of ERK1/2, JNK/SAPK, and v-akt murine thymoma viral oncongene (AKT) (serine 473) by immunoblotting. The expressions of the immediate early genes, c-fos and c-jun, were evaluated by reverse transcription polymerase chain reaction. RESULTS: Glucose at 10 mmol/L induced ERK1/2 and AKT phosphorylations and decreased SAPK/JNK phosphorylation. Palmitate (0.1 mmol/L) abolished the glucose effect on ERK1/2, AKT, and SAPK/JNK phosphorylations. LY294002 caused a similar effect. The inhibitory effect of palmitate on glucose induced ERK1/2 and AKT phosphorylation changes was not observed in the presence of fumonisin B1. Glucose increased c-fos and decreased c-jun expressions. Palmitate and LY294002 abolished these latter glucose effects. The presence of fumonisin B1 abolished the effect induced by palmitate on c-jun expression. CONCLUSIONS: Our results suggest that short-term changes of mitogen-activated protein kinase and AKT signaling pathways and c-fos and c-jun expressions caused by glucose are abolished by palmitate through phosphatidylinositol 3-kinase inhibition via ceramide synthesis. PMID- 19295453 TI - Follow-up study of K-ras mutations in the plasma of patients with pancreatic cancer: correlation with clinical features and carbohydrate antigen 19-9. AB - OBJECTIVE: We followed up the presence of Kirsten rat sarcoma (K-ras) mutations in plasma DNA and assessed its clinical value in patients with pancreatic cancer. METHODS: Plasma samples (N = 430) of 56 patients with pancreatic cancer and 13 patients with pancreatitis were analyzed by real-time polymerase chain reaction using peptide nucleic acid-mediated polymerase chain reaction clamping. RESULTS: K-ras mutations could be detected in the plasma DNA of 20 patients with cancer (36%). No K-ras mutation was found in the plasma of patients with pancreatitis. In 7 (35%) of 20 patients with lowly or moderately elevated carbohydrate antigen 19.9 (CA 19-9) levels lower than 100 U/mL, the result of the assay was positive for K-ras mutation. The combination of K-ras and CA 19-9 level determination gave a sensitivity for the diagnosis of pancreatic cancer of 91% (40/44) of the patients. Thirteen of 35 patients with pancreatic cancer (102 plasma samples) with elevated CA 19-9 levels (>35 U/mL) and altered K-ras gene showed significant correlation with elevated CA 19-9 levels (P=0.048). CONCLUSIONS: The summary of our approach of noninvasive, convenient, extremely high-sensitive K-ras mutation analysis in plasma might provide diagnostic and prognostic information to clinicians but will not be sufficient in a standardized early diagnosis of pancreatic carcinoma. The combination with CA 19-9 assay is useful for detection and prognostic evaluation of pancreatic carcinoma. PMID- 19295454 TI - Magnetic resonance cholangiopancreatography in asymptomatic pancreatic hyperenzymemia. AB - OBJECTIVES: To determine the type and frequency of pancreatic lesions detected by magnetic resonance cholangiopancreatography (MRCP) in subjects with asymptomatic pancreatic hyperenzymemia and to assess for a possible relationship between these lesions and the hyperenzymemia. METHODS: From January 2005 to May 2008, 63 subjects with asymptomatic pancreatic hyperenzymemia were studied by MRCP. In addition, amylase, pancreatic isoamylase, and lipase were determined for 5 consecutive days. RESULTS: In most subjects (n = 57, 90.5%), MRCP showed a normal pancreas. In the remaining 6 subjects (9.5%), the following alterations were found: pancreas divisum in 2, small intrapancreatic cyst in 2, anatomic variant of the Wirsung in 1, and mild dilatation of 3 secondary ducts in 1. In these 6 subjects, hyperenzymemia was highly variable from day to day, with frequent normalizations, as was also true for the 30 subjects with no MRCP alterations in whom diurnal enzyme determinations were made. CONCLUSIONS: Most of the subjects with asymptomatic pancreatic hyperenzymemia did not have pancreatic lesions detectable by MRCP. In the few subjects in whom a lesion was found, the great variability and the frequent transient normalization of serum enzyme levels tend to exclude a relation between the lesion and the hyperenzymemia. PMID- 19295455 TI - Early inhibition of prostaglandin synthesis by n-3 fatty acids determinates histologic severity of necrotizing pancreatitis. AB - OBJECTIVE: Previously, we observed decreased histopathological severity of acute necrotizing pancreatitis (ANP) by parenteral nutrition with n-3 fatty acids. Thus, we now sequentially analyzed the impact of n-3 fatty acids on prostaglandin and leukotriene synthesis in ANP. METHODS: One hundred ninety-eight Sprague Dawley rats (11 groups, n = 18) underwent intraductal glycodesoxycholat instillation and 6-hour cerulein infusion. Afterward, saline was infused in groups 2, 4, 6, 8, and 10, whereas groups 3, 5, 7, 9, and 11 received infusion rich in n-3 fatty acids (Omegaven, Fresenius Kabi, Bad Homburg, Germany). Animals were killed after 6 (group 1), 10 (groups 2 and 3), 14 (groups 4 and 5), 18 (groups 6 and 7), 22 (groups 8 and 9), and 26 hours (groups 10 and 11). The pancreas was histopathologically examined, and the pancreatic eicosanoid metabolism (prostaglandin E2, prostaglandin F1alpha [PGF1alpha], and leukotrienes) and lipid peroxidation (thiobarbituric acid-reactive substance, superoxide dismutase, and glutathione peroxidase) were analyzed. RESULTS: Between the 14th and 26th hours, histopathologic scores (edema, inflammation, bleeding, and necrosis) were reduced in the n-3 fatty acid group compared with the corresponding saline group. Pancreatic prostaglandin E2 and PGF1alpha were decreased between the 10th and 18th hour by n-3 fatty acids; PGF1alpha was reduced after 26 hours compared with the corresponding saline group. Lipid peroxidation was decreased by n-3 fatty acids after 14 hours (thiobarbituric acid reactive substance); however, there was no difference concerning lipid peroxidation protective enzymes (glutathione peroxidase and superoxide dismutase). CONCLUSIONS: Parenteral therapy with n-3 fatty acids decreased histopathologic severity in ANP by early inhibition of prostaglandin (E2 and F1alpha) synthesis and reduction of lipid peroxidation. PMID- 19295456 TI - Dexmedetomidine use in a pediatric cardiac intensive care unit: can we use it in infants after cardiac surgery? AB - OBJECTIVE: To assess clinical response of dexmedetomidine alone or in combination with conventional sedatives/analgesics after cardiac surgery. DESIGN: Retrospective study. SETTING: Pediatric cardiac intensive care unit. PATIENTS: Infants and neonates after cardiac surgery. MEASUREMENTS AND MAIN RESULTS: We identified 80 patients including 14 neonates, at mean age and weight of 4.1 +/- 3.1 months and 5.5 +/- 2 kg, respectively, who received dexmedetomidine for 25 +/ 13 hours at an average dose of 0.66 +/- 0.26 microgxkgxhr. Overall normal sleep to moderate sedation was documented 94% of the time and no pain to mild pain for 90%. Systolic blood pressure (SBP) decreased from 89 +/- 15 mm Hg to 85 +/- 11 mm Hg (p = .05), heart rate (HR) from 149 +/- 22 bpm to 129 +/- 16 bpm (p < .001), and respiratory rate (RR) remained unchanged. When baseline arterial blood gases were compared with the most abnormal values, pH decreased from 7.4 +/- 0.07 to 7.37 +/- 0.05 (p = .006), Po2 from 91 +/- 67 mm Hg to 66 +/- 29 mm Hg (p = .005), and CO2 increased from 45 +/- 8 mm Hg to 50 +/- 12 mm Hg (p = .001). At the beginning of the study, 37 patients (46%) were mechanically ventilated; and at 48 hours, 13 patients (16%) were still intubated and five patients failed extubation. Three groups of patients were identified: A, dexmedetomidine only (n = 20); B, dexmedetomidine with sedatives/analgesics (n = 38); and C, dexmedetomidine with both sedatives/analgesics and fentanyl infusion (n = 22). The doses of dexmedetomidine and rescue sedatives/analgesics were not significantly different among the three groups but duration of dexmedetomidine was longer in group C vs. A (p = .03) and C vs. B (p = .002). Pain, sedation, SBP, RR, and arterial blood gases were similar. HR was higher in group C vs. B (p = .01). Comparison between neonates and infants showed that infants required higher dexmedetomidine doses, 0.69 +/- 25 microgxkgxhr, and vs. 0.47 +/- 21 microgxkgxhr (p = .003) and had lower HR (p = .01), and RR (p = .009), and higher SBP (p < .001). CONCLUSIONS: Dexmedetomidine use in infants and neonates after cardiac surgery was well tolerated in both intubated and nonintubated patients. It provides an adequate level of sedation/analgesia either alone or in combination with low-dose conventional agents. PMID- 19295457 TI - Successful treatment with caspofungin of refractory Candida krusei candidemia in a very low birth weight preterm infant. PMID- 19295458 TI - Human metapneumovirus in Costa Rica. PMID- 19295459 TI - Respiratory syncytial virus and persistent infection of dendritic cells. PMID- 19295460 TI - Kaposi's sarcoma in the pediatric population: the critical need for a tissue diagnosis. AB - Kaposi's sarcoma (KS) is a low-grade vascular neoplasm mediated by the human herpesvirus-8. Only 1 clinical subtype, the endemic/African subtype, commonly affects the pediatric population. Although adults with KS often present with cutaneous findings and generalized lymphadenopathy, African children are more likely to present without classic skin findings. Definitive diagnosis requires histologic examination from tissue biopsy; however, as pathology resources are scarce in many developing African countries where KS is prominent, appropriate diagnosis and treatment of the condition are challenging. We report the case of a Malawian child who presented with generalized lymphadenopathy and was presumptively treated for lymphoma, with clinical worsening of his lesions. A diagnosis of KS was made after excisional biopsy of a superficial lymph node, with the initiation of appropriate therapy. The literature regarding pediatric KS is reviewed and recommendations are offered to allow accurate and timely diagnosis of the condition. PMID- 19295461 TI - False-negative Histoplasma antigen in acute pulmonary histoplasmosis: the value of urinary concentration by ultrafiltration and heat denaturation of serum proteins in detection of Histoplasma antigen. AB - We report an infant with localized pulmonary histoplasmosis in whom Histoplasma antibody assays, quantitative Histoplasma urine and serum antigen concentrations, and histopathologic findings of a mediastinal mass were nondiagnostic. A provisional diagnosis of histoplasmosis was established by using laboratory methods that increase the sensitivity of the antigen assay using ultrafiltration of urine and ethylenediaminetetraacetic acid/heat denaturation of serum proteins. PMID- 19295462 TI - Daptomycin therapy failure in an adolescent with methicillin-resistant Staphylococcus aureus bacteremia. AB - Daptomycin is a Food and Drug Administration-approved alternative to vancomycin for the treatment of serious methicillin-resistant Staphylococcus aureus infections in adults. Treatment failure with daptomycin is increasingly reported in adults, but data in children are limited. We report an adolescent patient with severe burn injuries who had persistent methicillin-resistant Staphylococcus aureus bacteremia and rapidly developed nonsusceptibility to daptomycin. We review the relevant literature. PMID- 19295463 TI - Prediction of Lyme meningitis based on a logistic regression model using clinical and cerebrospinal fluid analysis: a European study. AB - BACKGROUND: A prediction model based on clinical and cerebrospinal fluid (CSF) analysis has been proposed for the differentiation of Lyme meningitis (LM) from non-Lyme aseptic meningitis (NLAM) in the United States. No similar model has ever been proposed for European patients. The objective of our study was to develop a prediction model to differentiate LM from NLAM based on clinical and CSF biologic data. METHODS: The medical charts of all children older than 2 years of age admitted to our hospital from 1996 through 2006 with a definite diagnosis of LM were analyzed and compared retrospectively with those having a diagnosis of NLAM. Chart review included the duration of symptoms, the presence of cranial neuropathy, and CSF analysis. RESULTS: A total of 93 patients were included (LM: 26 patients; NLAM: 67 patients) in the study. Patients with LM had statistically more frequent cranial neuropathy (73% vs. 4%), displayed a longer duration of symptoms before admission (8.8 vs. 1.8 days), had a higher CSF protein (71 vs. 38 mg/d), and had a lower percentage of neutrophil cells in the CSF (3.4% vs. 51%) than patients with NLAM. A predicted probability was derived from these 4 variables. At a cutoff point of >0.432, the model had a negative predictive value of 100% and a positive predictive value of 92.3%, with a sensitivity of 100% and a specificity of 97%. CONCLUSIONS: We report the first European prediction model for LM. Owing to its high negative predictive value, this model may assist physicians in managing aseptic meningitis (AM) while awaiting serologic tests, especially in Lyme endemic regions. PMID- 19295464 TI - Clinical presentation of influenza in unselected children treated as outpatients. AB - BACKGROUND: Influenza causes a great disease burden on children especially in the outpatient setting. The signs and symptoms of influenza in unselected children treated as outpatients have not been previously published. METHODS: We assessed the clinical presentation of influenza in a prospective study of respiratory infections in preenrolled cohorts of children < or =13 years of age during 2 consecutive respiratory seasons (2231 child-seasons of follow-up). We examined the children and obtained a nasal swab for the detection of influenza during every episode of illness, regardless of the presence or absence of fever or the severity of the symptoms. RESULTS: Influenza was virologically confirmed in 372 children, of whom 353 (95%) providing complete data on the signs and symptoms were included in the analyses. A total of 95% of these children were febrile, and 50% had fever > or =39.0 degrees C. Among children <3 years of age, 20% had fever > or =40.0 degrees C. Seventy-seven percent of the children had cough and 78% had rhinitis. In children 7 to 13 years of age, only 39% had headache and 13% had myalgia. CONCLUSIONS: High fever is a prominent sign of influenza in children, and the clinical presentation of influenza is most severe in children <3 years of age. Headache and myalgia are not typical features of influenza in outpatient children. Most children with influenza have rhinitis during the early phase of the illness, which makes the clinical diagnosis of influenza difficult especially in the youngest children. PMID- 19295465 TI - Current recommended dosing of vancomycin for children with invasive methicillin resistant Staphylococcus aureus infections is inadequate. AB - BACKGROUND: Vancomycin area-under-the-concentration-time-curve (AUC) for 24 hours divided by the minimum inhibitory concentration (MIC) (AUC24/MIC) >400 optimally treats invasive methicillin-resistant Staphylococcus aureus (MRSA) infections in adults. It is unknown whether recommended vancomycin dosing regimens for children achieve this value. METHODS: AUC24/MIC was calculated in children using vancomycin doses of 40 and 60 mg/kg/d. AUC24 was calculated as daily dose/vancomycin clearance. Vancomycin clearance in children was estimated by 2 approaches: (1) previously literature-reported vancomycin clearance, and (2) calculated vancomycin clearance using previously derived predictor models and a hypothetical population of healthy children. Representative MIC of hospital MRSA isolates was used (0.5, 1.0, and 2.0 microg/mL). RESULTS: The MIC50/90 for pediatric MRSA isolates in the previous year was 1.0 microg/mL. With a dose of 40 mg/kg/d, both approaches consistently predicted AUC24/MIC <400 when MIC was 1.0 microg/mL. At 60 mg/kg/d, AUC24/MIC >400 was more readily achieved when MIC was 1.0 microg/mL, however, an MIC of 2.0 microg/mL resulted in AUC24/MIC <400 for both dosing regimens. CONCLUSIONS: A vancomycin dose of 40 mg/kg/d in children is unlikely to achieve the recommended pharmacodynamic target of AUC24/MIC >400 for invasive MRSA infections even when MIC is 1.0 microg/mL. A starting dose of 60 mg/kg/d should be used in settings where isolates with MIC of 1.0 are common. Alternatives to vancomycin should strongly be considered for patients with MIC > or =2.0 microg/mL. PMID- 19295466 TI - The effect of universal toddlers-only hepatitis A virus vaccination program on seropositivity rate in unvaccinated toddlers: evidence for reduced virus circulation in the community. AB - BACKGROUND: In July 1999, a national toddler-only hepatitis A virus (HAV) vaccination program was introduced in Israel. Passive and active surveillance showed a large reduction in disease rate, but an objective measurement was needed. We hypothesized that toddler's vaccination in a population living in an endemic area would reduce virus circulation, resulting in reduced HAV seropositivity rates in unvaccinated toddlers. METHODS: The study was conducted among Bedouin children in southern Israel, for whom HAV vaccine coverage reached 85.5% and 74.9% for first and second HAV vaccine doses, respectively, in 2000. Toddlers received 2 doses of HAV vaccine at 18 and 24 months. Data on vaccine coverage was received from well-baby clinics. Sera were obtained from healthy unvaccinated 16- to 20-month-old toddlers. Anti-HAV immunoglobulin (Ig)G concentrations were tested by enzyme-linked immunosorbent assay. RESULTS: A total of 629 sera were tested (209 obtained in 1991-2000 and 420 obtained in 2001 2002). Seropositivity rates of > or =100 mIU/mL ranged from 16.2% to 19.6% in 1991 through 2000 (children born before immunization program). These rates dropped to 2% in 2001-2002 and to 0% in 2003 through 2007. Furthermore, IgG concentrations were significantly lower (P < 0.001) in samples taken in 2000, only a few months after beginning of vaccination, than in those taken before initiation of the HAV immunization program (1991-1998), suggesting a marked reduction in circulating HAV resulting in natural boosting. CONCLUSIONS: Because HAV vaccines are licensed in children > or =12 months old, rates of anti-HAV seropositivity in unvaccinated toddlers can be an objective and sensitive tool to evaluate the effect of immunization program on virus circulation. This method is of special value in communities where no appropriate surveillance is in place. PMID- 19295467 TI - Prevalence of HPV-DNA and anti-HPV antibodies in women from Girardot, Colombia. AB - OBJECTIVE: To assess the frequency of HPV-DNA detection, human papillomavirus (HPV) seropositivity, presence of cervical lesions, and its relationship with certain socio-demographic factors in women from Girardot, Colombia from 2006 to 2007. METHODS: Nine hundred fifty-three women attending their regular Pap smear control voluntarily provided cervical cells and blood samples for HPV-DNA analysis and ELISA detection of anti-L1 peptides and virus-like particles (VLPs) antibodies after answering a questionnaire regarding sexual behaviors, number of births, smoking habits, and socio-demographic background. RESULTS: Twenty-six of the 953 women being examined (2.73%) presented cervical cell abnormalities. A frequency of 36.62% (95% CI: 33.52%-39.7%) HPV seropositivity was detected with peptide 18301, 35.36% (95% CI: 32.3%-38.4%) with 18283, and 32.95% (95% CI: 29.9% 36%) with 18294, whereas VLPs detected a 43% seropositivity (95% CI: 39.8% 46.2%). Antibody frequency found with all peptides was significantly higher in women having cervical abnormalities (atypical squamous cells of undetermined significance and high-grade squamous intraepithelial lesions) compared with those having normal cytologies. Peptide 18283 reported a significantly higher seropositivity (35.71%) in women >44 years old, whereas peptides 18301 and 18294 evidenced a significantly lower seropositivity in those who had never given birth. HR-HPV-DNA was detected in 157 (20.50%) of 766 cervical samples amplifying positively for the beta-globin housekeeping gene. CONCLUSION: Peptides 18283, 18294, and 18301 were more specific and more sensitive than VLPs for detecting women with HR-HPV-DNA positive cervical lesions. Therefore, they could be useful in the design of a serological test for detecting HR-HPV-infected women having cervical lesions at a risk of progressing to cervical cancer. PMID- 19295468 TI - Cervical human papillomavirus incidence and persistence in a cohort of HIV negative women in Zimbabwe. AB - BACKGROUND: Persistent infections with oncogenic human papillomavirus (HPV) types are causally related to cervical cancer. Little is known about the distribution of HPV types, independent risk factors of incidence and persistence, and patterns of persistence in sub-Saharan Africa. METHODS: A cohort of 2040 Zimbabwean women was enrolled in a randomized trial assessing the effect of diaphragm/gel provision on human immunodeficiency virus and HPV acquisition. Data from the study arms were pooled for this analysis because diaphragm/gel provision did not affect HPV acquisition and clearance. Clinicians collected cervical samples for HPV testing at enrollment, 12 months, and exit (median 21 months). RESULTS: HPV prevalence was 24.5% for any HPV type and 16.1% for oncogenic types. HPV incidence at 12 months was 23.3% for any HPV type and 11.4% for oncogenic types. HPV58 had the highest baseline prevalence (5.0%) and incidence (2.4%). Type specific persistence was 29.8% among all HPV infections over a median of 21 months of follow-up. Baseline predictors of incident HPV infection were younger age, having more than 1 lifetime sexual partner, infrequent condom use, herpes simplex virus-2 positive serology, and having a sexually transmissible infection or a different HPV type at enrollment. Baseline predictors of persistent HPV infection were younger age, having more than 1 lifetime sexual partner, and having a high-risk partner. CONCLUSIONS: The novel association between herpes simplex virus-2 seropositivity and incident HPV infection warrants further investigation. Having a high-risk partner is a potentially modifiable risk factor for persistent HPV infection. The relatively high prevalence of HPV58 has implications for vaccine development. PMID- 19295469 TI - High resolution imaging of epithelial injury in the sheep cervicovaginal tract: a promising model for testing safety of candidate microbicides. AB - BACKGROUND: Access to readily available large animal models and sensitive noninvasive techniques that can be used for the evaluation of microbicide-induced changes in tissue could significantly facilitate preclinical evaluations of microbicide safety. The sheep cervicovaginal tract, with stratified squamous epithelium similar to humans, holds promise as a large animal model used before nonhuman primates. In addition, optical coherence tomography (OCT) could enable high resolution visualization of tissue morphology and noninvasive assessment of microbicide-induced epithelial injury. METHODS: We evaluated the dose response of sheep cervicovaginal tract to benzalkonium chloride (BZK). Twenty sheep received treatment with phosphate-buffered saline or BZK solution (2%, 0.2%, or 0.02%). Pre- and posttreatment colposcopy and OCT images were collected and graded based on World Health Organization criteria and a previously reported scoring system, respectively. Biopsies were collected and the degree of epithelial injury and its thickness was assessed based on histology and OCT. RESULTS: The sheep cervicovagina exhibited anatomic and microscopic features similar to the human. Extensive loss of the epithelium was noted on colposcopy and OCT after application of 2% BZK. Colposcopy detected findings in half of sheep and OCT in all sheep treated with 0.2% BZK. OCT detected differences in the 0.02% BZK treated group compared with controls, whereas colposcopy failed to detect any changes. CONCLUSIONS: The sheep cervicovagina is similar to humans, and exhibits dose dependent epithelial changes after BZK treatment. These findings suggest that the sheep model and OCT may become valuable tools for the safety evaluation of candidate microbicides, and warrant continued development. PMID- 19295470 TI - HIV risk among bisexually and homosexually active racially diverse young men. AB - OBJECTIVES: We addressed gaps in current HIV prevention research by examining the differences between 2 groups of young men: men who have sex with men only (MSM/O) and men who have sex with men and women (MSM/W). We examined patterns and correlates of sexual risk, and considered how race/ethnicity may affect these relationships. METHODS: Cross-sectional self-report data were collected from a racially diverse sample of 10,295 young MSM from 1999 to 2002. The sample comprised data from 13 urban locations across the US. RESULTS: MSM/W reported less unprotected anal intercourse (UAI) than MSM/O, despite reporting less exposure to HIV prevention interventions, lower social support, and less awareness of antiretroviral therapies. African American men were more likely to be MSM/W and less likely to report UAI. Ever getting an HIV test was associated with less UAI only among African American participants (MSM/W or MSM/O) in racial/ethnic group-specific analyses. CONCLUSIONS: HIV prevention interventions for MSM should address differences between MSM/W and MSM/O. An important component of HIV prevention efforts designed for African American MSM/W and MSM/O should be HIV testing. PMID- 19295471 TI - The economic burden of pediculosis pubis and scabies infections treated on an outpatient basis in the United States: evidence from private insurance claims data, 2001-2005. PMID- 19295472 TI - Incidence of HIV and sexually transmitted infections and risk factors for acquisition among young methamphetamine users in northern Thailand. AB - BACKGROUND: Southeast Asia is experiencing an epidemic of methamphetamine use, a drug associated with risky sexual behaviors, putting a large segment of the population at increased risk for sexually transmitted infections (STIs) and HIV and in need of prevention efforts. Incidence estimates of STIs are rare in Southeast Asia, especially among newer risk groups. STUDY DESIGN: We enrolled methamphetamine users aged 18 to 25 years in a 12-month randomized behavioral intervention trial in Chiang Mai, Thailand in 2005. Behavioral questionnaires were administered at visits every 3 months, and biologic specimens were collected at baseline and 12 months to test for common STIs (chlamydia, gonorrhea, HSV-2, and HIV). Poisson regression with robust variance was used to determine risk factors for incident STIs. RESULTS: Overall, 12.7% of 519 participants acquired at least 1 STI. Chlamydia was the most common (10.6%), followed by HSV-2 (4.0%), gonorrhea (2.9%), and HIV (0.6%). Risk factors for both men and women included self-reported incarceration and having a casual sex partner during follow-up, and having a prevalent STI at baseline. Additionally, among women, having 2 or more heterosexual partners, and among men, having a greater frequency of drunkenness were risk factors for STI acquisition. CONCLUSIONS: Although HIV incidence is low in this population, incidence of other STIs is high compared with previous studies of young Thai adults. Risk factors for acquisition emphasize the need for new prevention strategies targeted toward current populations at risk. PMID- 19295473 TI - Central cannabinoid 1 receptor antagonist administration prevents endotoxic hypotension affecting norepinephrine release in the preoptic anterior hypothalamic area. AB - It is widely assumed that LPS lowers arterial pressure during sepsis by stimulating release of TNF-alpha and other vasoactive mediators from macrophages. However, recent data from this and other laboratories have shown that LPS hypotension can be prevented by inhibiting afferent impulse flow in the vagus nerve, by blocking neuronal activity in the nucleus of the solitary tract, or by blocking alpha-adrenergic receptors in the preoptic area/anterior hypothalamic area (POA). These findings suggest that the inflammatory signal is conveyed from the periphery to the brain via the vagus nerve, and that endotoxic shock is mediated through a central mechanism that requires activation of POA neurons. In the present study, we tested whether central cannabinoid 1 (CB1) receptors participate in the control of arterial pressure during endotoxemia based on evidence that hypothalamic neurons express CB1 receptors and synthesize the endogenous CB anandamide. We found that intracerebroventricular administration of rimonabant, a CB1 receptor antagonist, inhibited the fall in arterial pressure evoked by LPS significantly in both conscious and anesthetized rats. Rimonabant attenuated both the immediate fall in arterial pressure evoked by LPS and the second, delayed hypotensive phase that leads to tissue ischemia and death. Rimonabant also prevented the associated LPS-induced rise in extracellular fluid norepinephrine concentrations in the POA. Furthermore, rimonabant attenuated the associated increase in plasma TNF-alpha concentrations characteristic of the late phase of endotoxic hypotension. These data indicate that central CB1 receptors may play an important role in the initiation of endotoxic hypotension. PMID- 19295474 TI - Cardiovascular collapse and vascular permeability changes in an ovine model of methicillin-resistant Staphylococcus aureus sepsis. AB - Methicillin-resistant Staphylococcus aureus (MRSA) infections with severe outcomes such as sepsis and septic shock are progressively increasing in both the community and in hospital settings. We hypothesized that overexpression of reactive nitrogen and oxygen species and vascular endothelial growth factor (VEGF) play a pivotal role in cardiovascular collapse associated with vascular hyperpermeability in MRSA sepsis. Twelve sheep were surgically prepared and randomized into a control (noninjured; n = 6) and a sepsis (injured; n = 6) group. Animals in the sepsis group were subjected to cotton smoke inhalation and instillation of 2.5 x 10(11) colony-forming units of live MRSA into both lungs. Cardiovascular variables in the control group remained stable, whereas the MRSA sepsis group developed a hypotensive and hyperdynamic circulatory shock state beginning at 6 h associated with significantly increased vascular permeability evidenced by increased prefemoral lymph flow starting at 12 h and permeability index from 12 to 18 h, higher fluid accumulation from 12 to 24 h, and significantly decreased plasma protein concentration and oncotic pressure beginning at 6 h compared with control animals. Myocardial 3-nitrotyrosine (3-NT) protein, poly (adenosine diphosphate-ribose), and VEGF mRNA expressions measured after the 24-h experiment were significantly increased in the injured animals as well. These results evidence that excessive production of reactive radicals and VEGF may play a major role in cardiovascular collapse and vascular hyperpermeability in MRSA sepsis. PMID- 19295475 TI - Rosmarinic acid protects against experimental sepsis by inhibiting proinflammatory factor release and ameliorating hemodynamics. AB - The present study was to investigate the effects of rosmarinic acid (RA) in cultured RAW264.7 cells and experimental model of sepsis induced by cecal ligation and puncture in rats and the potential mechanism. Results showed that RA concentration dependently down-regulated the levels of TNF-alpha, IL-6, and high mobility group box 1 protein in LPS-induced RAW264.7 cells, inhibited the IkappaB kinase pathway, and modulated nuclear factor-kappaB. Intravenous injection of RA alone or in combination with imipenem reduced cecal ligation and puncture-induced lethality in rats. In addition, serum levels of TNF-alpha, IL-6, high-mobility group box 1 protein, triggering receptor expressed on myeloid cells, and endotoxin were down-regulated; in contrast, serum level of IL-10 was up regulated. Amelioration of hemodynamics and decrease in serum enzyme activities and myeloperoxidase in lung, liver, and small intestine were also observed after RA injection. These data indicate that the antisepsis effect of RA was mediated by decreasing local and systemic levels of a wide spectrum of inflammatory mediators. This article provides the first evidence that RA has the capacity to inactivate inflammatory response in sepsis. The anti-inflammatory mechanism of RA may inhibit activation of the nuclear factor- kappaB pathway by inhibiting IkappaB kinase activity. PMID- 19295476 TI - Liver X receptor agonist GW3965 dose-dependently regulates lps-mediated liver injury and modulates posttranscriptional TNF-alpha production and p38 mitogen activated protein kinase activation in liver macrophages. AB - Modulation of the host inflammatory response to infection may be a key approach to improve the outcome of patients with sepsis and organ injury. We previously reported that pretreatment of rats with the liver X receptor (LXR) agonist GW3965 reduced the liver injury associated with endotoxemia and attenuated the production of TNF-alpha by rat Kupffer cells. Here, we examine the dose-dependent effect of GW3965 on liver injury and cytokine production in a rat model of endotoxemia and explore the mechanisms underlying TNF-alpha attenuation in Kupffer cells. Low doses of GW3965 (0.1 or 0.3 mg/kg) administered 30 min before infusion of LPS and peptidoglycan significantly attenuated the increase in plasma levels of the liver injury markers alanine aminotransferase and bilirubin (6 h) as well as the inflammatory mediators TNF-alpha (1 h) and prostaglandin E2 (6 h) associated with endotoxemia. In contrast, pretreatment with a higher dose of GW3965 (1.0 mg/kg) had no such effect. Studies in primary cultures of rat Kupffer cells demonstrated that LXR agonist treatment attenuated both the secreted and cell-associated levels of TNF-alpha, whereas TNF-alpha mRNA levels were not altered. Phosphorylated p38 mitogen-activated protein kinase, which plays a major role in production of TNF-alpha at the posttranscriptional level, was attenuated by GW3965 treatment in Kupffer cells. Experiments in murine LXR-deficient Kupffer cells demonstrated enhanced production of TNF-alpha in Kupffer cells from LXR alpha(-/-) mice when challenged with LPS compared with LXR-beta(-/-) and wild type Kupffer cells. Taken together, these results argue in favor of a novel mechanism for LXR-mediated attenuation of liver injury by interfering with posttranscriptional regulation of TNF-alpha in Kupffer cells. PMID- 19295477 TI - Protective effect of suberoylanilide hydroxamic acid against LPS-induced septic shock in rodents. AB - We have recently found that suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, improves survival in a lethal model of hemorrhagic shock in rats. The purpose of the present study was to determine whether SAHA treatment would prevent LPS-induced septic shock and improve the survival in a murine model. C57BL/6J mice were randomly divided into two groups. Experimental mice were given intraperitoneal SAHA (50 mg/kg) in vehicle dimethyl sulfoxide fluid (n = 10). The control mice (n = 10) received vehicle dimethyl sulfoxide only. They were injected with LPS (20 mg/kg, i.p.) 2 h later, and the animals from the treatment group were given a second dose of SAHA. Survival was monitored during the next 7 days. In a parallel study, mice treated with or without SAHA were subjected to LPS insult while normal (sham) mice serviced as controls. 1) Lungs were harvested at 3 and 48 h for analysis of gene expression and pathologic changes, respectively; 2) spleens were isolated for analysis of neutrophilic cell population. In addition, RAW264.7 mouse macrophages were cultured to assess the effects of SAHA on LPS-induced inflammation in vitro. All mice in the control group that were subjected to LPS challenge died in less than 48 h. However, SAHA treated animals displayed a significantly higher 1-week survival rate (87.5%) compared with the control group (0%). Moreover, LPS insult decreased the acetylation of histone proteins (H2A, H2B, and H3), elevated the levels of TNF alpha in vivo (circulation) and in vitro (culture medium), increased the neutrophilic cell population in the spleen, enhanced the expression of TNF-alpha and IL-1beta genes in lung tissue, and augmented the pulmonary neutrophil infiltration. In contrast, SAHA treatment markedly attenuated all of these LPS induced alterations. We report for the first time that administration of SAHA (50 mg/kg) significantly attenuates a variety of inflammatory markers and improves long-term survival after a lethal LPS insult. PMID- 19295478 TI - Enoxaparin attenuates endothelial damage with less bleeding compared with unfractionated heparin in endotoxemic rats. AB - Prophylactic use of anticoagulants during sepsis is strongly recommended for the prevention of venous thrombosis. Moreover, recent studies suggested the positive effects of anticoagulants to the inflammation. In this study, we planned to confirm the effects of heparins on protecting against endothelial damage in endotoxemia. In addition, we also examined the differences between unfractionated heparin (UFH) and enoxaparin. Wistar rats received 8.5 mg/kg (i.v.) LPS, followed by a bolus infusion of either 350 U/kg of UFH, 2.0 mg/kg of enoxaparin, or placebo. Microscopic observation of the mesenteric microcirculation and the measurement of the bleeding area after puncture with a microneedle were performed 3 h later (n = 6 in each group). In another series, blood samples were taken 3 h after the LPS injection, and blood cell counts, coagulation markers, and organ damage markers were measured (n = 6 in each). As a result, the leukocyte adherence to the endothelium was significantly reduced in both the UFH and enoxaparin groups, and thus, endothelial damage was attenuated in these groups. The bleeding area was markedly expanded in the UFH group compared with the other groups (P < 0.01 each). The decrease in white blood cells and platelet count was significantly suppressed in the enoxaparin group compared with the UFH group (P < 0.05 each). The fibrinogen level was maintained at significantly better levels, and the elevation of alanine aminotransferase was significantly suppressed in enoxaparin group (P < 0.05 each). In conclusion, both UFH and enoxaparin protect against endothelial damage by preventing leukocyte adhesion. However, UFH significantly increases the bleeding area, whereas enoxaparin does not increase bleeding, and thus, it can reduce organ damages in the endotoxemic rat. PMID- 19295479 TI - Gamma-globulin levels in patients with community-acquired septic shock. AB - Polyclonal intravenous immunoglobulins (IVIGs) can modulate the host immune response and may improve outcomes in some patients. In this prospective, noninterventional study, we assessed the time-course of gamma-globulin concentrations in 21 patients with septic shock and evaluated the relationship of gamma-globulin concentrations to disease severity and outcome. Six patients (28.5%) died. Sixteen patients (76%) had hypo-gamma-globulinemia at admission: 12 (57%) had low IgG concentrations (G650 mg/dL), 2 had low IgA concentrations (G70 mg/dL), and 9 had low IgM concentrations (G40 mg/dL). Two patients had low concentrations of all three gamma-globulins; these patients both died from refractory shock within 2 days. Patients with low IgG concentrations were indistinguishable at baseline from patients with normal IgG concentrations but had fewer vasopressor-free days (over 28 days) and more frequently developed acute lung injury/acute respiratory distress syndrome (3/9 vs. 10/12; P = 0.02). All deaths occurred in the patients with low IgG concentrations (6/ 12 vs. 0/9). There was a variable increase in kappa and lambda free light chains, a marker of gamma-globulin synthesis, over time with no significant difference between low and normal IgG groups. This pilot study indicates that low concentrations of gamma-globulins, especially IgG, are common in patients with community-acquired septic shock and persist over time even when sepsis resolves. Despite similar presentation, patients with hypo-IgG had greater vasopressor requirements, were more likely to develop acute lung injury/acute respiratory distress syndrome, and had higher mortality. Patients with low IgG concentrations may represent a logical target group to study the effects of Ig supplementation in septic shock. PMID- 19295480 TI - Systemic inflammation increases intestinal permeability during experimental human endotoxemia. AB - Although the gut is often considered the motor of sepsis, the relation between systemic inflammation and intestinal permeability in humans is not clear. We analyzed intestinal permeability during experimental endotoxemia in humans. Before and during experimental endotoxemia (Escherichia coli LPS, 2 ng/kg), using polyethylene glycol (PEG) as a permeability marker, intestinal permeability was analyzed in 14 healthy subjects. Enterocyte damage was determined by intestinal fatty acid binding protein. Endotoxemia induced an inflammatory response. Urinary PEGs 1,500 and 4,000 recovery increased from 38.8 +/- 6.3 to 63.1 +/- 12.5 and from 0.58 +/- 0.31 to 3.11 +/- 0.93 mg, respectively (P < 0.05). Intestinal fatty acid binding protein excretion was not affected by endotoxemia. The peak serum IL 10 concentrations correlated with the increase in PEG 1,500 recovery (r = 0.48, P = 0.027). Systemic inflammation results in an increased intestinal permeability. The increase in intestinal permeability is most likely caused by inflammation induced paracellular permeability, rather than ischemia-mediated enterocyte damage. PMID- 19295481 TI - Edaravone, a novel free radical scavenger, reduces high-mobility group box 1 and prolongs survival in a neonatal sepsis model. AB - Free radicals play an important role in the inflammatory process of sepsis. We hypothesized that edaravone, a novel free radical scavenger, can suppress pathophysiological events and prolong survival in a neonatal sepsis cecal ligation and perforation (CLP) model. Of 32 3-day-old anesthetized and mechanically ventilated piglets, 11 received CLP only, 10 received CLP and edaravone treatment starting 30 min after CLP, and 11 constituted a sham (control) group. Mean arterial pressure (MAP), heart rate, cardiac output, arterial blood gas, serum total hydroperoxide, nitrite and nitrate, TNF-alpha, and high-mobility group box 1 (HMGB1) were measured before CLP and at 1, 3, and 6 h after CLP. Compared with the CLP group, the edaravone group showed higher MAP at 6 h, lower heart rate at 1 and 3 h, lower total hydroperoxide at 1 h, lower nitrite and nitrate at 3 and 6 h, and higher (although not significantly so) mean cardiac output at 1, 3, and 6 h. TNF-alpha elevation was delayed from 1 h in the CLP group to 3 h in the edaravone group. In the edaravone group, HMGB1 did not change significantly at any time, whereas in the CLP group, it increased at 6 h. Survival times were longer in the edaravone group than in the CLP group (15.4 +/- 1.4 vs. 10.2 +/- 1 h; P < 0.005). In addition, each of the serial dilutions of edaravone had a higher biological antioxidant potential than tempol does. In conclusion, edaravone suppressed free radicals, delayed the TNF-alpha surge, and prevented HMGB1 elevation, thereby maintaining MAP and prolonging survival time in a neonatal sepsis CLP model. PMID- 19295482 TI - Effects of TNF-alpha-converting enzyme inhibition on acute lung injury induced by endotoxin in the rat. AB - We studied the effects of TNF-converting enzyme inhibition with Y-41654, which down-regulates the production of soluble TNF-alpha (sTNF-alpha), on acute lung injury induced by intratracheal administration of LPS. We first verified in vitro that pretreatment of isolated alveolar macrophages from Sprague-Dawley male rats with 20 microL of 0.1-mM Y-41654, decreased significantly (P < 0.05) the concentration of sTNF-alpha in cell supernatants induced by 10 microg/mL of LPS. We then studied four groups of rats (each n = 10) including 1) a control group, 2) an LPS group (300 microg /kg, instilled intratracheally), 3) a Y-41654 group, and 4) a treatment group treated with Y-41654 after LPS instillation. Y-41654, 10 mg/kg in 0.7 mL of phosphate-buffered saline, was administered (i.v.), 15 min before and 0.5, 1.5, 2.5, and 3.5 h after saline or LPS instillation. The animals were observed for 4 h. In the animals treated with Y-41654, the concentrations of sTNF-alpha and protein in bronchoalveolar lavage fluid, and the number of neutrophils in lung tissue and bronchoalveolar lavage fluid were significantly lower at 4 h than in the LPS group (P < 0.05). In conclusion, sTNF-alpha plays an important role in the development of acute lung injury induced by intratracheal administration of LPS, in part modulating neutrophil kinetics. PMID- 19295483 TI - A longer duration of polymyxin B-immobilized fiber column hemoperfusion improves pulmonary oxygenation in patients with septic shock. AB - Endotoxin plays an important role in the pathogenesis of septic shock. Exposure of endothelial cells to endotoxin activates endothelial cells and increases the surface expression of adhesion molecules, markers of endothelial damage in organ dysfunction. Endotoxin adsorption therapy by polymyxin B-immobilized fiber column (PMX) hemoperfusion has been used for the treatment of septic shock patients. In this study, we measured plasma concentrations of endotoxin and soluble adhesion molecules in septic shock patients before and after the PMX treatment then observed on the relationships between actual duration of use and various outcomes. Sixteen patients with septic shock were studied. The 28-day mortality rate was 50%. The elevated plasma concentrations of endotoxin decreased after the PMX treatment in the survivors but not in the nonsurvivors. The norepinephrine dose and plasma concentrations of soluble endothelial leukocyte adhesion molecule 1 and soluble intercellular adhesion molecule 1 significantly (P < 0.05) decreased in the PMX greater-than-2-h (prolonged) group than in the PMX 2-h (conventional) group (-17.8 +/- 14.6 vs. -1.8 +/- 2.7 microg/min, -143.0 +/- 111.0 vs. 0 +/- 2.8 ng/mL, and -126.2 +/- 144.9 vs. 16.5 +/- 108.1 ng/mL, respectively). Changes in the PaO2-FiO2 ratio and the Sequential Organ Failure Assessment score were significantly (P < 0.05) more improved in the PMX greater than-2-h group than in the PMX 2-h group (75.4 +/- 80.7 vs. 1.2 +/- 49.2 and -0.8 +/- 1.8 vs. 2.2 +/- 1.9 torr, respectively). We thus suggest that a longer duration of PMX treatment may improve the pulmonary oxygenation associated with decreased adhesion molecules in septic shock. PMID- 19295484 TI - Inhibition of c-Jun N-terminal kinase after hemorrhage but before resuscitation mitigates hepatic damage and inflammatory response in male rats. AB - Inhibition of c-Jun N-terminal kinase (JNK) by a cell-penetrating, protease resistant JNK peptide (D-JNKI-1) before hemorrhage and resuscitation (H/R) ameliorated the H/R-induced hepatic injury and blunted the proinflammatory changes. Here we tested the hypothesis if JNK inhibition at a later time point after hemorrhagic shock but before the onset of resuscitation-in a rat model of H/R also confers protection. Twenty-four male Sprague-Dawley rats (250 - 350 g) were randomly divided into 4 groups: 2 groups of shock animals were hemorrhaged to a MAP of 32 to 37 mmHg for 60 min and randomly received either D-JNKI-1 (11 mg/kg i.p.) or sterile saline as vehicle immediately before the onset of resuscitation. Two groups of sham-operated animals underwent surgical procedures without H/R and were either D-JNKI-1 or vehicle treated. Rats were killed 2 h later. Serum activity of alanine aminotransferase and serum lactate dehydrogenase after H/R increased 3.5-fold in vehicle-treated rats as compared with D-JNKI-1 treated rats. Histopathological analysis revealed that hepatic necrosis and apoptosis (hematoxylin-eosin, TUNEL, and M30, respectively) were significantly inhibited in D-JNKI-1-treated rats after H/R. Hepatic oxidative (4 hydroxynonenal) and nitrosative (3-nitrotyrosine) stress as well as markers of inflammation (hepatic and serum IL-6 levels and hepatic infiltration with polymorphonuclear leukocytes) were also reduced in D-JNKI-1-treated rats. LPS stimulated TNF-alpha release from whole blood from hemorrhaged and resuscitated animals was higher in vehicle-treated rats as compared with D-JNKI-1-treated rats. c-Jun N-terminal kinase inhibition after hemorrhage before resuscitation resulted in a reduced activation of c-Jun. Taken together, these results indicate that D-JNKI-1 application after hemorrhagic shock before resuscitation blunts hepatic damage and proinflammatory changes during resuscitation. Hence, JNK inhibition is even protective when initiated after blood loss before resuscitation. These experimental results indicate that the JNK pathway may be a possible treatment option for the harmful consequences of H/R. PMID- 19295485 TI - Effects of carvedilol on mortality and inflammatory responses to severe hemorrhagic shock in rats. AB - The nonselective beta-adrenoceptor and the selective alpha1-adrenoceptor blocker carvedilol are widely used in hypertensive and/or cardiac failure patients because of its efficacy. However, there have been few studies regarding the effects of carvedilol on severe hemorrhagic shock. The present study was performed to evaluate the effects of carvedilol on severe hemorrhagic shock in rats. Twenty-four male Sprague-Dawley rats were randomly assigned to 1 of the following 2 groups (n = 12 per group): control group, no medication; and treatment group, oral administration of carvedilol (10 mg/kg per day) for 5 days. All animals were anesthetized with i.p. pentobarbital. Severe hemorrhagic shock was induced by partial exsanguination. Eight minutes after shock, all removed blood was returned to the animal. No other treatments were administered before, during, or after shock. Hemodynamics and arterial blood gases were recorded, mortality was calculated for the 5-h observation period, and plasma cytokine concentrations were measured at 5 h after shock. The mortality rates at 5 h after cardiac arrest were 8% and 50% for control and treatment groups, respectively. The increases in base deficit and lactate concentrations were less in the control group than that in the treatment group. Moreover, the increases in TNF-alpha concentrations were less in the control group than in the treatment group. The present study indicated that oral administration of carvedilol had adverse effects on mortality and inflammatory responses to severe hemorrhagic shock in rats. These findings suggest that carvedilol may adversely affect recovery from severe hemorrhagic shock. PMID- 19295486 TI - The effect of antithrombin on pulmonary endothelial damage induced by crush injury. AB - After crush injury, patients often experience multiple organ dysfunction syndrome. In this study, we focused on vascular endothelial damage, which is believed to be a possible cause of multiple organ dysfunction syndrome, and revealed a pathological condition of distant organ failure. In particular, the lung is an especially prone target organ at the time of systemic inflammatory invasion after crush injury. We ascertained the effect of antithrombin (AT), which has recently attracted attention for its endothelial protective effects. Using a rat model of crush syndrome, we assessed severity of systemic inflammation and vascular endothelial damage through a blood test and degree of lung injury and centrally focused on morphological analysis of endothelium over time. Crush injury significantly elevated the blood concentration of tissue plasminogen activator-plasminogen activator inhibitor 1 complex, monocyte chemoattractant protein 1, and IL-6. Accumulation of active inflammatory cells (OX-42-positive cells) and expression of von Willebrand factor and vascular cell adhesion molecule 1 significantly increased in the lung 24 h after releasing crush. After 48 h, disarray of alveolar structure and alveolar hemorrhage appeared. Antithrombin administration significantly suppressed accumulation of inflammatory cells, expression of von Willebrand factor and vascular cell adhesion molecule 1, and mortality rate. Our research demonstrates that crush injury induces acute lung injury as distant organ failure, and it would seem that AT administration diminishes vascular endothelial damage and is effective against crush injury. PMID- 19295488 TI - The selective adenosine A2A receptor agonist CGS 21680 reduces JNK MAPK activation in oligodendrocytes in injured spinal cord. AB - Permanent functional deficit after spinal cord injury (SCI) arises from both mechanical injury and from secondary tissue reactions involving inflammation. Adenosine is an important regulator of inflammatory mechanisms. Although functional studies indicate a protective effect of adenosine A2A receptor agonists in SCI, the basic molecular mechanisms accounting for the their protective effects from SCI have to be fully elucidated. In this study, we investigated if the selective A2A receptor agonist 2-[p-(2-carboxyethyl) phenethylamino]-5'-N-ethylcarboxamidoadenosine (CGS 21680) administered after SCI has protective effects against tissue damage, motor deficit, and different inflammatory readouts. Spinal cord injury was induced in mice by extradural compression of a section of the SC exposed via a four-level T5-T8 laminectomy. CGS 21680, administered by subcutaneously implanted osmotic minipumps after SCI, clearly reduced motor deficit for up to 19 days after operation. The drug repeatedly administered intraperitoneally after SCI reduced tissue damage, influx of myeloperoxidase-positive leukocytes, nuclear factor-kappaB activation and iNOS expression in injured spinal cord tissue 24 h after SCI. Enhanced immunoreactivity of microglia, astrocytes, and oligodendrocytes (stained by anti CD11/B, anti-glial fibrillary acidic protein, and anti-Olig2 antibodies, respectively) was also observed 24 h after SCI. Neurons lose immunoreactivity in the nucleus. c-Jun amino-terminal kinase (JNK) mitogen-activated protein kinase, quantified by Western blot, was definitely activated in injured tissue. CGS 21680 treatment significantly reduced JNK phosporylation. Phospho-JNK mitogen-activated protein kinase was de novo expressed selectively in oligodendrocytes. CGS 21680 reduced phospho-JNK immunostaining in oligodendrocytes. Data indicate that protection by the A2A agonist is secondary to reduced leukocyte recruitment in the damaged area. A reducing effect of JNK activation in oligodendrocytes might account for protective effect of the A2A agonist against SCI-induced demyelination. PMID- 19295490 TI - Evolution of portal-systemic collateral vasopressin response in endotoxemic portal hypertensive rats. AB - Cirrhotic patients with portal hypertension and variceal hemorrhage are vulnerable to endotoxemia. However, the direct influence of endotoxemia on portal systemic collateral vasculature remains unexplored. In this study, portal hypertension was induced in Sprague-Dawley rats by partial portal vein ligation. On the 7th day after portal vein ligation, at 0.5, 1.5, and 5 h post endotoxin (LPS; Escherichia coli serotype O111:B4, 3 mg/kg, i.p., E0.5, E1.5 and E5, respectively) or saline (control, C0.5, C1.5, and C5, respectively) injection, hemodynamic measurements and concentration-response relationships to arginine vasopressin (AVP; 10(-10)-10(-7) mol/L) in collateral vascular bed were obtained. In another six parallel groups, reverse-transcriptase-polymerase chain reaction of iNOS, eNOS, and endothelin 1 (ET-1) mRNA expressions for splenorenal shunt, the most prominent intra-abdominal collateral vessel, was performed. The results showed that E0.5 had lower perfusion pressure changes to AVP and higher splenorenal shunt eNOS expression than C0.5 group (P < 0.05). Compared with C1.5, tachycardia, higher perfusion pressure changes and enhanced splenorenal shunt iNOS and ET-1 expression were observed in E1.5 group (P < 0.05). In E5, systemic and portal hypotension with markedly enhanced collateral AVP responsiveness and splenorenal shunt iNOS and ET-1 expressions were noted (P < 0.05). In conclusion, vasoactive substances counterregulation participates, at least in part, the time dependent changes of collateral AVP responsiveness in endotoxemic portal hypertensive rats. PMID- 19295489 TI - Elevated glucose and fatty acid levels impair substance P-induced dermal microvascular endothelial cell migration and proliferation in an agarose gel model system. AB - Substance P (SP), a sensory nerve derived neuropeptide, has been implicated in wound repair. Our hypothesis was that oxidative effects of elevated glucose and fatty acid levels as seen with diabetes mellitus inhibit SP-mediated endothelial cell directional migration and proliferation. Using a 2% agarose gel, immortalized human microvascular endothelial cells (HMEC-1) were plated into a 1.5-mm well, and agonist (SP; 10(-4) mol/L) was loaded into a 3-mm well; controls included NaCl, albumin (bovine serum albumin), and vascular endothelial cell growth factor. The SP receptor antagonist spantide 1 was used to confirm SP specificity. Elevated glucose (40 mmol/L) and fatty acids (40 micromol/L) were added to the medium with and without vitamin E and vitamin C treatment to determine whether endothelial cell responses to SP were altered by metabolic perturbations and whether they could be recovered with antioxidant treatment. Using computer-assisted image analysis, migration distance was measured. Cells were counted using a hemocytometer. Human microvascular endothelial cell 1 migration toward the SP exceeded NaCl or bovine serum albumin; vascular endothelial cell growth factor had similar effects. The SP receptor antagonist, spantide, inhibited SP-induced HMEC-1 migration. Substance P treatment was associated with increased cell number. Ki-67 staining was increased in SP-treated cells compared with controls. Elevated glucose and fatty acid levels diminished cell migration toward SP. The antioxidants vitamins C and E significantly improved proliferation but only marginally improved migration. Our data suggest that glucose and fatty acids perturb SP-induced HMEC-1 migration and proliferation in an agarose gel migration model. PMID- 19295491 TI - Different patterns of Siglec-9-mediated neutrophil death responses in septic shock. AB - Sialic-acid-binding immunoglobulin-like lectin (Siglec) 9 mediates death signals in neutrophils. The objective of this study was to determine the heterogeneity of neutrophil death responses in septic shock patients and to analyze whether these ex vivo data are related to the severity and outcome of septic shock. In this prospective cohort study, blood samples of patients with septic shock (n = 26) in a medical-surgical intensive care unit (ICU) were taken within 24 h of starting the treatment of septic shock (phase A), after circulatory stabilization (phase B), and 10 days after admission or at ICU discharge if earlier (phase C). Neutrophil death was quantified in the presence and absence of an agonistic anti Siglec-9 antibody after 24 h ex vivo. In phase A, two distinct patterns of Siglec 9-mediated neutrophil death were observed: resistance to neutrophil death (n = 14; Siglec-9 nonresponders) and increased neutrophil death (n = 12; Siglec-9 responders) after Siglec-9 ligation compared with neutrophils from normal donors. Experiments using a pharmacological pan-caspase-inhibitor provided evidence for caspase-independent neutrophil death in Siglec-9 responders upon Siglec-9 ligation. There were no differences between Siglec-9 responders and nonresponders in length of ICU or hospital stay of survivors or severity of organ dysfunction. Taken together, septic shock patients exhibit different ex vivo death responses of blood neutrophils after Siglec-9 ligation early in shock. Both the resistance and the increased susceptibility to Siglec-9-mediated neutrophil death tend to normalize within 72 h after shock. Further studies are required to understand the role of Siglec-9-mediated neutrophil death in septic shock. PMID- 19295492 TI - The effect of iloprost on renal dysfunction after renal I/R using cystatin C and beta2-microglobulin monitoring. AB - The purpose of this study was to investigate the effect of iloprost, a cytoprotective prostacyclin analog, on renal injury during unilateral renal I/R in rats and to determine whether the levels of serum cystatin C (CyC) and beta2 microglobulin (B2M), as markers of glomerular function, might denote this injury. Thirty-two Wistar rats were randomized into four groups (n = 8) as follows: control (sham laparotomy), renal I/R (60-min left renal ischemia and 120-min reperfusion), renal I/R + iloprost (20 ng kg(-1) min(-1) infusion during renal I/R period, i.v.), and control + iloprost. Blood and kidney tissue samples were obtained for biochemical and histological analysis from all rats. Serum urea, creatinine, CyC, and B2M levels were evaluated for biochemical analysis. Histopathological changes in renal structure were examined for histological analysis. Serum urea, creatinine, and CyC levels were significantly increased in the renal I/R group. Iloprost treatment decreased these three markers in the renal I/R + iloprost group. beta2-Microglobulin levels were not significantly changed in any group. Histological analyses showed that renal I/R elicited significant renal injury, whereas iloprost significantly decreased I/R-induced renal injury. Serum CyC level is one of the good indicators of acute renal damage due to I/R produced by renal artery occlusion. In contrast, we have shown that there are no significant changes in the levels of serum B2M levels that would make it an accurate diagnostic tool for detecting acute changes in renal injury subject to renal I/R in rats. PMID- 19295493 TI - Activated protein C can be used as a prophylactic as well as a therapeutic agent for heat stroke in rodents. AB - The present study was attempted to assess the prophylactic and the therapeutic effect of human recombinant activated protein C (APC; drotrecogin-alpha, activated) in experimental heat stroke. Anesthetized rats were divided into two groups and given vehicle solution 1 h before the start or immediately after the termination of heat stress (isotonic sodium chloride solution, 2 mL kg(-1) of body weight, i.v.) or APC (1-10 mg in 2 mL of isotonic sodium chloride solution per kilogram of body weight, i.v.). They were exposed to ambient temperature of 40 degrees C for 100 min to induce heat stroke. When the vehicle-pretreated rats underwent heat stress, their survival time values were found to be 57 to 71 min. Pretreatment or treatment with APC significantly increased survival time (122-221 min). All vehicle-pretreated heat stroke animals displayed systemic inflammation (evidenced by increased TNF-alpha, IL-1alpha, and IL-6) and activated coagulation (evidenced by increased levels of activated partial thromboplastin time, prothrombin time, and D-dimer and decreased levels of both platelet count and protein C). Biochemical assay also revealed that both renal and hepatic dysfunction (e.g., increased plasma levels of blood urea nitrogen, creatinine, adenine aminotransferase, aspartate aminotransferase, and alkaline phosphatase) were noted during heat stroke. A significant decrease in both cerebral blood flow and partial pressure of oxygen in hypothalamus were also observed in vehicle pretreated heat stroke animals. These heat stroke reactions were all significantly reduced by pretreatment or treatment with human recombinant APC. The results indicate that human recombinant APC can be used as a prophylactic and a therapeutic agent for experimental heat stroke by ameliorating systemic inflammation, hypercoagulable state, and multiple organ dysfunction. PMID- 19295494 TI - Hematopoietic stem cell proliferation modeling under the influence of hematopoietic-inducing agent. AB - The process by which hematopoietic stem cells (HSCs) residing in the bone marrow differentiate into blood cells is known as hematopoiesis. In the event of hemorrhagic shock, it is crucial for the HSC to rapidly differentiate into new committed erythroid progenitor cells that will give rise to erythrocytes. Growth factors and cytokines enhance the self-renewing process of HSC and are therefore crucial to restoring normal levels of blood cells in the body. Hematopoietic inducing agents (HIAs) such as the cytokine erythropoietin and granulocyte-colony stimulating factor play a vital role in hematopoiesis because they are capable of inducing the proliferation of stem cells. The aim of the current study is to mathematically model the effect of HIA on the proliferation rate of hematopoietic stem cells at varying levels of oxygenation. The role of HIA was analyzed by constructing a set of coupled ordinary differential equations upon which mathematical analysis was performed. The model makes predictions of hematopoietic activity during low oxygen levels (ranging from 3% to 15%) similar to conditions ranging from acute blood loss to normal conditions. PMID- 19295495 TI - Endotoxin and interferon-gamma inhibit translation in skeletal muscle cells by stimulating nitric oxide synthase activity. AB - The purpose of the present study was to test the hypothesis that endogenous NO negatively affects translation in skeletal muscle cells after exposure to a combination of endotoxin (LPS) and interferon-gamma (IFN-gamma). Individually, LPS and IFN-gamma did not alter protein synthesis, but in combination, they inhibited protein synthesis by 80% in C2C12 myotubes. The combination of LPS and IFN-gamma dramatically downregulated the autophosphorylation of the mammalian target of rapamycin and its substrates S6K1 and 4EBP-1. The phosphorylation of ribosomal protein S6 was decreased, whereas phosphorylation of elongation factor 2 and raptor was enhanced, consistent with defects in both translation initiation and elongation. Reduced S6 phosphorylation occurred 8 to 18 h after LPS/IFN-gamma and coincided with a prolonged upregulation of NOS2 messenger RNA and protein. NOS2 protein expression and the LPS/IFN-gamma-induced fall in phosphorylated S6 were prevented by the proteasome inhibitor MG-132. The general NOS inhibitor, L NAME, and the specific NOS2 inhibitor, 1400W, also prevented the LPS/IFN-gamma induced decrease in protein synthesis and restored translational signaling. LPS/IFN-gamma downregulated the phosphorylation of multiple Akt substrates, including the proline-rich Akt substrate 40, while enhancing the phosphorylation of raptor on a 5'-AMP-activated kinase (AMPK)-regulated site. The negative effects of LPS/IFN-gamma were blunted by the AMPK inhibitor compound C. The data suggest that, in combination, LPS and IFN-gamma induce a prolonged expression of NOS2 and excessive production of NO that reciprocally alter Akt and AMPK activity and consequently downregulate translation via reduced mammalian target of rapamycin signaling. PMID- 19295496 TI - Acute liver injury and biomarkers: a biological lesson from indocyanine green. PMID- 19295497 TI - Homing of hematopoietic cells to the bone marrow. AB - Homing is the phenomenon whereby transplanted hematopoietic cells are able to travel to and engraft or establish residence in the bone marrow. Various chemomkines and receptors are involved in the homing of hematopoietic stem cells. This paper outlines the classic homing protocol used in hematopoietic stem cell studies. In general this involves isolating the cell population whose homing needs to be investigated, staining this population with a dye of interest and injecting these cells into the blood stream of a recipient animal. The recipient animal is then sacrificed at a pre-determined time after injection and the bone marrow evaluated for the percentage or absolute number of cells which are positive for the dye of interest. In one of the most common experimental schemes, the homing efficiency of hematopoietic cells from two genetically distinct animals (a wild type animal and the corresponding knock-out) is compared. This article describes the hematopoietic cell homing protocol in the framework of such as experiment. PMID- 19295498 TI - A set of surgical chain saws. PMID- 19295499 TI - A boost in microRNAs shapes up the neuron. PMID- 19295500 TI - Long-range signalling in activation of the translational GTPase EF-Tu. PMID- 19295501 TI - Decreased renal expression of vascular endothelial growth factor in lupus nephritis is associated with worse prognosis. AB - Recent studies suggest that vascular endothelial growth factor (VEGF) plays a crucial role in the preservation of renal function and may also serve as a useful biomarker in monitoring the progression of lupus nephritis (LN). Here we sought to correlate intrarenal VEGF expression with renal histopathology and prognosis of LN. Biopsy specimens from 35 patients with Class III or IV LN (ISN/RPS categorization) were found to have lower levels of intrarenal VEGF than those found in biopsy tissue taken from 10 donor kidneys sampled at the time of allograft reperfusion. This reduced amount of VEGF mRNA in the patients with LN negatively correlated with glomerular endocapillary proliferation, crescent formation, and a high histologic activity index but was positively associated with increased numbers of urinary podocytes. The level of intrarenal VEGF mRNA accurately predicted the deterioration of renal function in these patients within 12 months. Our study shows that expression of VEGF in renal tissue may serve as a molecular marker of renal damage and may be a predictive factor for short-term loss of kidney function in patients with LN. PMID- 19295502 TI - Metabolic abnormalities are present in adults with elevated serum cystatin C. AB - Although metabolic anomalies are often seen in advanced chronic kidney disease (CKD), their presence in more mild states is unknown. We studied 6722 participants in the Third National Health and Nutrition Examination Survey, dividing them into three mutually exclusive groups consisting of those having a normal or mildly reduced estimated glomerular filtration rate (eGFR) (Modification of Diet in Renal Disease (MDRD) formula), those with normal or elevated serum cystatin C, and those with clinically relevant moderate or severely reduced eGFR (stage 3 or 4 of CKD). The prevalence of several metabolic abnormalities associated with moderate to advanced CKD was determined after standardization for age, race-ethnicity, and gender. In the absence of stage 3 or 4 CKD, patients with elevated serum cystatin C had a higher prevalence of low hemoglobin and elevated uric acid, homocysteine, phosphorus, fibrinogen, and C reactive protein than patients with a normal serum cystatin C. Our results show that in adults with normal or mildly reduced eGFR, elevated serum cystatin C is associated with an increased prevalence of metabolic abnormalities traditionally found in moderate or severe CKD. Elevated serum cystatin C may identify patients with 'preclinical' kidney disease not detected by traditional serum creatinine measurements. PMID- 19295503 TI - Methods for female contraception: a model for innovation in drug delivery systems. PMID- 19295504 TI - Predictive biomarkers in the development of oncology drugs: a therapeutic industry perspective. PMID- 19295505 TI - Are we optimizing gestational diabetes treatment with glyburide? The pharmacologic basis for better clinical practice. AB - Glyburide's pharmacokinetics (PK) and pharmacodynamics have not been studied in women with gestational diabetes mellitus (GDM). The objective of this study was to assess steady-state PK of glyburide, as well as insulin sensitivity, beta-cell responsivity, and overall disposition indices after a mixed-meal tolerance test (MMTT) in women with GDM (n = 40), nonpregnant women with type 2 diabetes mellitus (T2DM) (n = 26), and healthy pregnant women (n = 40, MMTT only). At equivalent doses, glyburide plasma concentrations were approximately 50% lower in pregnant women than in nonpregnant subjects. The average umbilical cord/maternal plasma glyburide concentration ratio at the time of delivery was 0.7 +/- 0.4. Insulin sensitivity was approximately fivefold lower in women with GDM as compared with healthy pregnant women. Despite comparable beta-cell responsivity indices, the average beta-cell function corrected for insulin resistance was more than 3.5-fold lower in women with glyburide-treated GDM than in healthy pregnant women. Women with GDM in whom glyburide treatment has failed may benefit from alternative medication or dosage escalation; however, fetal safety should be kept in mind. PMID- 19295506 TI - Personalized dosing of cyclophosphamide in the total body irradiation cyclophosphamide conditioning regimen: a phase II trial in patients with hematologic malignancy. AB - This study investigates the efficacy and safety of personalized cyclophosphamide (CY) dosing in 50 patients receiving CY along with total body irradiation (TBI). Participants received CY 45 mg/kg with subsequent therapeutic drug monitoring using Bayesian parameter estimation to personalize the second CY dose to a target area under the curve (AUC) for carboxyethylphosphoramide mustard (CEPM) (a reporter molecule for CY-derived toxins) and for hydroxycyclophosphamide (to ensure engraftment). The mean second CY dose was 66 mg/kg; the total dose ranged from 45 to 145 mg/kg. After completion of this phase II study, we compared participants' clinical outcomes with those of concurrent controls (n = 100) who received TBI along with standard CY doses of 120 mg/kg. Patients receiving personalized CY dosing had significantly lower postconditioning peak total serum bilirubin (P = 0.03); a 38% reduction in the hazard of acute kidney injury (AKI) (P = 0.03); and nonrelapse and overall survival rates similar to those in the controls (P = 0.70 and 0.63, respectively) despite the lower doses of CY administered to most of the patients in the personalized dosage group. PMID- 19295507 TI - mGluR5 positive allosteric modulators facilitate both hippocampal LTP and LTD and enhance spatial learning. AB - Highly selective positive allosteric modulators (PAMs) of metabotropic glutamate receptor subtype 5 (mGluR5) have emerged as a potential approach to treat positive symptoms associated with schizophrenia. mGluR5 plays an important role in both long-term potentiation (LTP) and long-term depression (LTD), suggesting that mGluR5 PAMs may also have utility in improving impaired cognitive function. However, if mGluR5 PAMs shift the balance of LTP and LTD or induce a state in which afferent activity induces lasting changes in synaptic function that are not appropriate for a given pattern of activity, this could disrupt rather than enhance cognitive function. We determined the effect of selective mGluR5 PAMs on the induction of LTP and LTD at the Schaffer collateral-CA1 synapse in the hippocampus. mGluR5-selective PAMs significantly enhanced threshold theta-burst stimulation (TBS)-induced LTP. In addition, mGluR5 PAMs enhanced both DHPG induced LTD and LTD induced by the delivery of paired-pulse low-frequency stimulation. Selective potentiation of mGluR5 had no effect on LTP induced by suprathreshold TBS or saturated LTP. The finding that potentiation of mGluR5 mediated responses to stimulation of glutamatergic afferents enhances both LTP and LTD and supports the hypothesis that the activation of mGluR5 by endogenous glutamate contributes to both forms of plasticity. Furthermore, two systemically active mGluR5 PAMs enhanced performance in the Morris water maze, a measure of hippocampus-dependent spatial learning. Discovery of small molecules that enhance both LTP and LTD in an activity-appropriate manner shows a unique action on synaptic plasticity that may provide a novel approach for the treatment of impaired cognitive function. PMID- 19295508 TI - Bidirectional effects of fentanyl on dendritic spines and AMPA receptors depend upon the internalization of mu opioid receptors. AB - Fentanyl is a frequently used and abused opioid analgesic and can cause internalization of mu opioid receptors (MORs). Receptor internalization modulates the signaling pathways of opioid receptors. As changes in dendritic spines and synaptic AMPA receptors play important roles in addiction and memory loss, we investigated how fentanyl affects dendritic spines and synaptic AMPA receptors in cultured hippocampal neurons. Fentanyl at low concentrations (0.01 and 0.1 microM) caused the collapse of dendritic spines and decreased the number of AMPA receptor clusters. In contrast, fentanyl at high concentrations (1 and 10 microM) had opposite effects, inducing the emergence of new spines and increasing the number of AMPA receptor clusters. These dose-dependent bidirectional effects of fentanyl were blocked by a selective MOR antagonist CTOP at 5 microM. In neurons that had been transfected with HA-tagged or GFP-tagged MORs, fentanyl at high concentrations induced persistent and robust internalization of MORs, whereas fentanyl at lower concentrations induced little or transient receptor internalization. The blockade of receptor internalization with the expression of dominant-negative Dynamin I (the K44E mutant) reversed the effect of fentanyl at high concentrations, supporting a role of receptor internalization in modulating the dose-dependent effects of fentanyl. In contrast to morphine, the effects of fentanyl on dendritic spines are distinctively bidirectional and concentration dependent, probably due to its ability to induce robust internalization of MORs at high concentrations. The characterization of the effects of fentanyl on spines and AMPA receptors may help us understand the roles of MOR internalization in addiction and cognitive deficits. PMID- 19295509 TI - Genetic and clinical predictors of sexual dysfunction in citalopram-treated depressed patients. AB - Sexual dysfunction is a major contributor to treatment discontinuation and nonadherence among patients treated with selective serotonin reuptake inhibitors (SSRIs). The mechanisms by which depressive symptoms in general, as well as SSRI exposure in particular, may worsen sexual function are not known. We examined genetic polymorphisms, including those of the serotonin and glutamate systems, for association with erectile dysfunction, anorgasmia, and decreased libido during citalopram treatment. Clinical data were drawn from a nested case-control cohort derived from the STAR(*)D study, a multicenter, prospective, effectiveness trial in outpatients with nonpsychotic major depressive disorder (MDD). Self reports of erectile dysfunction, decreased libido, or difficulty achieving orgasm based on the Patient-Rated Inventory of Side Effects were examined among Caucasian subjects (n=1473) for whom DNA and adverse effect measures were available, and who were treated openly with citalopram for up to 14 weeks. Of 1473 participants, 799 (54%) reported decreased libido; 525 (36%) reported difficulty achieving orgasm. Of 574 men, 211 (37%) reported erectile dysfunction. Using a set-based test for association, single nucleotide polymorphisms in glutamatergic genes were associated with decreased libido (GRIA3; GRIK2), difficulty achieving orgasm (GRIA1), and difficulty achieving erection (GRIN3A) (experiment-wide permuted p<0.05 for each). Evidence of association persisted after adjustment for baseline clinical and sociodemographic differences. Likewise, evidence of association was similar when the cohort was limited to those who did not report a given adverse event at the first post-baseline visit (ie, those whose adverse events were known to be treatment emergent). These hypothesis-generating analyses suggest the potential for glutamatergic treatment targets for sexual dysfunction during major depressive episodes. PMID- 19295510 TI - Neuronal correlates of brain-derived neurotrophic factor Val66Met polymorphism and morphometric abnormalities in bipolar disorder. AB - The brain-derived neurotrophic factor (BDNF) Val66Met polymorphism has been proposed as a possible candidate for involvement in the pathophysiology of bipolar disorder (BD). To determine whether an association exists between the BDNF Val66Met genotype and morphometric abnormalities of the brain regions involved in memory and learning in BD and healthy subjects. Forty-two BD patients and 42 healthy subjects were studied. Interactions between BDNF Val66Met genotype and diagnosis in gray (GM) volumes were analyzed using an optimized voxel-based morphometry technique. Declarative memory function was assessed with the California Verbal Learning Test II. Left and right anterior cingulate GM volumes showed a significant interaction between genotype and diagnosis such that anterior cingulate GM volumes were significantly smaller in the Val/Met BD patients compared with the Val/Val BD patients (left P=0.01, right P=0.01). Within-group comparisons revealed that the Val/Met carriers showed smaller GM volumes of the dorsolateral prefrontal cortex compared with the Val/Val subjects within the BD patient (P=0.01) and healthy groups (left P=0.03, right P=0.03). The Val/Met healthy subjects had smaller GM volumes of the left hippocampus compared with the Val/Val healthy subjects (P<0.01). There was a significant main effect of diagnosis on memory function (P=0.04), but no interaction between diagnosis and genotype was found (P=0.48). The findings support an association between the BDNF Val66Met genotype and differential gray matter content in brain structures, and suggest that the variation in this gene may play a more prominent role in brain structure differences in subjects affected with BD. PMID- 19295511 TI - Will the Kraepelinian dichotomy survive DSM-V? AB - Kraepelin proposed dementia praecox and manic-depressive illness as the two major psychotic disorders. This paradigm is still prevalent, but observations of overlapping boundaries between bipolar disorder and schizophrenia challenge this dichotomy. However, the concept of schizophrenia has been radically altered from the original Kraepelinian proposal. We defend the two psychoses positions, but suggest two flaws in the heuristic application: (1) overlapping features, such as psychotic symptoms, are not decisive in differential diagnosis; and (2) each disorder is a syndrome, not a disease entity. An alternative paradigm based on domains of pathology is more powerful for studies of etiology, pathophysiology, and therapeutic discovery. PMID- 19295512 TI - STAT3 inhibition of gluconeogenesis is downregulated by SirT1. AB - The fasting-activated longevity protein sirtuin 1 (SirT1, ref. 1) promotes gluconeogenesis in part, by increasing transcription of the key gluconeogenic genes pepck1 and g6pase, through deacetylating PGC-1alpha and FOXO1 (ref. 4). In contrast, signal transducer and activator of transcription 3 (STAT3) inhibits glucose production by suppressing expression of these genes. It is not known whether the inhibition of gluconeogenesis by STAT3 is controlled by metabolic regulation. Here we show that STAT3 phosphorylation and function in the liver were tightly regulated by the nutritional status of an animal, through SirT1 mediated deacetylation of key STAT3 lysine sites. The importance of the SirT1 STAT3 pathway in the regulation of gluconeogenesis was verified in STAT3 deficient mice in which the dynamic regulation of gluconeogenic genes by nutritional status was disrupted. Our results reveal a new nutrient sensing pathway through which SirT1 suppresses the inhibitory effect of STAT3, while activating the stimulatory effect of PGC-1alpha and FOXO1 on gluconeogenesis, thus ensuring maximal activation of gluconeogenic gene transcription. The connection between acetylation and phosphorylation of STAT3 implies that STAT3 may have an important role in other cellular processes that involve SirT1. PMID- 19295513 TI - Modularity of MAP kinases allows deformation of their signalling pathways. AB - Eukaryotic protein kinase pathways have both grown in number and changed their network architecture during evolution. We wondered if there are pivotal proteins in these pathways that have been repeatedly responsible for forming new connections through evolution, thus changing the topology of the network; and if so, whether the underlying properties of these proteins could be exploited to re engineer and rewire these pathways. We addressed these questions in the context of the mitogen-activated protein kinase (MAPK) pathways. MAPK proteins were found to have repeatedly acquired new specificities and interaction partners during evolution, suggesting that these proteins are pivotal in the kinase network. Using the MAPKs Fus3 and Hog1 of the Saccharomyces cerevisiae mating and hyper osmolar pathways, respectively, we show that these pivotal proteins can be re designed to achieve a wide variety of changes in the input-output properties of the MAPK network. Through an analysis of our experimental results and of the sequence and structure of these proteins, we show that rewiring of the network is possible due to the underlying modular design of the MAPKs. We discuss the implications of our findings on the radiation of MAPKs through evolution and on how these proteins achieve their specificity. PMID- 19295514 TI - Histone modifications at human enhancers reflect global cell-type-specific gene expression. AB - The human body is composed of diverse cell types with distinct functions. Although it is known that lineage specification depends on cell-specific gene expression, which in turn is driven by promoters, enhancers, insulators and other cis-regulatory DNA sequences for each gene, the relative roles of these regulatory elements in this process are not clear. We have previously developed a chromatin-immunoprecipitation-based microarray method (ChIP-chip) to locate promoters, enhancers and insulators in the human genome. Here we use the same approach to identify these elements in multiple cell types and investigate their roles in cell-type-specific gene expression. We observed that the chromatin state at promoters and CTCF-binding at insulators is largely invariant across diverse cell types. In contrast, enhancers are marked with highly cell-type-specific histone modification patterns, strongly correlate to cell-type-specific gene expression programs on a global scale, and are functionally active in a cell-type specific manner. Our results define over 55,000 potential transcriptional enhancers in the human genome, significantly expanding the current catalogue of human enhancers and highlighting the role of these elements in cell-type-specific gene expression. PMID- 19295515 TI - Temporally precise in vivo control of intracellular signalling. AB - In the study of complex mammalian behaviours, technological limitations have prevented spatiotemporally precise control over intracellular signalling processes. Here we report the development of a versatile family of genetically encoded optical tools ('optoXRs') that leverage common structure-function relationships among G-protein-coupled receptors (GPCRs) to recruit and control, with high spatiotemporal precision, receptor-initiated biochemical signalling pathways. In particular, we have developed and characterized two optoXRs that selectively recruit distinct, targeted signalling pathways in response to light. The two optoXRs exerted opposing effects on spike firing in nucleus accumbens in vivo, and precisely timed optoXR photostimulation in nucleus accumbens by itself sufficed to drive conditioned place preference in freely moving mice. The optoXR approach allows testing of hypotheses regarding the causal impact of biochemical signalling in behaving mammals, in a targetable and temporally precise manner. PMID- 19295516 TI - Directional Delta and Notch trafficking in Sara endosomes during asymmetric cell division. AB - Endocytosis has a crucial role during Notch signalling after the asymmetric division of fly sensory organ precursors (SOPs): directional signalling is mediated by differential endocytosis of the ligand Delta and the Notch effector Sanpodo in one of the SOP daughters, pIIb. Here we show a new mechanism of directional signalling on the basis of the trafficking of Delta and Notch molecules already internalized in the SOP and subsequently targeted to the other daughter cell, pIIa. Internalized Delta and Notch traffic to an endosome marked by the protein Sara. During SOP mitosis, Sara endosomes containing Notch and Delta move to the central spindle and then to pIIa. Subsequently, in pIIa (but not in pIIb) Notch appears cleaved in Sara endosomes in a gamma-secretase- and Delta internalization-dependent manner, indicating that the release of the intracellular Notch tail to activate Notch target genes has occurred. We thus uncover a new mechanism to bias signalling even before asymmetric endocytosis of Sanpodo and Delta takes place in the daughter cells: already during SOP mitosis, asymmetric targeting of Delta and Notch-containing Sara endosomes will increase Notch signalling in pIIa and decrease it in pIIb. PMID- 19295518 TI - This time. PMID- 19295519 TI - From students to scientists at XLAB. PMID- 19295520 TI - Chemicals turn human embryonic stem cells towards beta cells. PMID- 19295521 TI - Getting a handle on protein prenylation. PMID- 19295522 TI - Glycome 'fingerprints' provide definitive clues to HIV origins. PMID- 19295523 TI - Back to the future of nucleic acid self-amplification. PMID- 19295524 TI - Hypoxia sensing goes gauche. PMID- 19295529 TI - Pharmacogenetics of pharmacoecology: which route to personalized medicine? PMID- 19295533 TI - Searching where the light is worse: overemphasizing genes and underplaying environment in the quest to reduce substance misuse. PMID- 19295526 TI - Emerging methods for the production of homogeneous human glycoproteins. AB - Most circulating human proteins exist as heterogeneously glycosylated variants (glycoforms) of an otherwise homogeneous polypeptide. Though glycan heterogeneity is most likely important to glycoprotein function, the preparation of homogeneous glycoforms is important both for the study of the consequences of glycosylation and for therapeutic purposes. This review details selected approaches to the production of homogeneous human N- and O-linked glycoproteins with human-type glycans. Particular emphasis is placed on recent developments in the engineering of glycosylation pathways within yeast and bacteria for in vivo production, and on the in vitro remodeling of glycoproteins by enzymatic means. The future of this field is very exciting. PMID- 19295534 TI - Genes and addictions. PMID- 19295535 TI - Stimulation of cholecystokinin-A receptors with Gl181771X: a failed clinical trial that did not test the pharmacogenetic hypothesis for reduction of food intake. AB - There are two interacting components in a clinical trial: the drug and the study design. When a trial does not work, we blame the drug--and the study is usually not published. This Commentary provides a context for the use of efficacy pipeline pharmacogenetics (PGx) in therapeutic programs. Jordan et al. published the results of an obesity trial with a cholecystokinin-A (CCK-A) receptor agonist and concluded that CCK-A by itself does not have a central role in long-term energy balance. The conclusions were sound, the report accurate, and the journal commendable for publishing a negative study, but the trial design was misdirected -it did not build on phase IIA information and did not test the proposed mechanism of action. The hypotheses should have been based on the original putative role of a central mechanism affecting appetite, which had been validated using efficacy PGx in phase IIA. PMID- 19295536 TI - Evaluating QTc in patients with advanced Parkinson's disease: overcoming artifact. AB - Electrocardiograms (ECGs) in patients with Parkinson's disease are affected by artifacts related to muscle tremor. Malik et al. report methods used in QTc study in patients with Parkinson's disease that markedly reduce the noise and variance of QTc in a sample of ECGs that are undeniably difficult to interpret. This study adds significant experience and novel methods that have the potential to further enhance the evaluation of the effect of a drug on QTc. PMID- 19295537 TI - The Dutch vision of clinical pharmacology. AB - Recent position papers addressing the profession of clinical pharmacology have expressed concerns about the decline of interest in the field among clinicians and medical educators in the United Kingdom and other Western countries, whether clinical pharmacology is actually therapeutics, and whether the profession should be limited to physicians. The Dutch Society for Clinical Pharmacology and Biopharmacy offers answers to these questions and presents a new model for clinical pharmacology. PMID- 19295540 TI - Transplantation immunobiology: two important themes. PMID- 19295541 TI - Extended IL10 haplotypes and their association with HIV progression to AIDS. AB - Interleukin-10 (IL-10) is a pleiotropic cytokine with both immunosuppressive and immunostimulatory functions. Its roles in infections and autoimmunity may have resulted in selective pressures on polymorphisms within the gene, leading to genomic coexistence of several semi-conserved haplotypes involved with diverse pathogen interactions during genomic evolution. Previous studies focused either exclusively on promoter haplotypes or on individual SNPs. We genotyped 21 single nucleotide polymorphisms in the human IL10 gene and examined this variation compared to other mammalian species sequences. Haplotype heterogeneity in human populations is centered around 'classic' 'proximal' promoter polymorphisms: -592, -819 and -1082. High-producing GCC haplotypes are by far the most numerous and diverse group, the intermediate IL-10 producing ACC-inclusive haplotypes seem to be related most closely to the ancestral haplotype, and the ATA-inclusive haplotypes cluster a separate branch with strong bootstrap support. We looked at associations of corresponding haplotypes with HIV progression. A haplotype trend regression confirmed that individuals carrying the low-producing ATA-inclusive haplotypes in European Americans progress to AIDS faster, and most likely explain the role of IL10. Our findings are consistent with the hypothesis that existing polymorphisms in this gene may reflect a balance of historic adaptive responses to autoimmune, infectious and other disease agents. PMID- 19295542 TI - Reproducible association with type 1 diabetes in the extended class I region of the major histocompatibility complex. AB - The high-risk human leukocyte antigen (HLA)-DRB1, DQA1 and DQB1 alleles cannot explain the entire type 1 diabetes (T1D) association observed within the extended major histocompatibility complex. We have earlier identified an association with D6S2223, located 2.3 Mb telomeric of HLA-A, on the DRB1(*)03-DQA1(*)0501 DQB1(*)0201 haplotype, and this study aimed to fine-map the associated region also on the DRB1(*)0401-DQA1(*)03-DQB1(*)0302 haplotype, characterized by less extensive linkage disequilibrium. To exclude associations secondary to DRB1-DQA1 DQB1 haplotypes, 205 families with at least one parent homozygous for these loci, were genotyped for 137 polymorphisms. We found novel associations on the DRB1(*)0401-DQA1(*)03-DQB1(*)0302 haplotypic background with eight single nucleotide polymorphisms (SNPs) located within or near the PRSS16 gene. In addition, association at the butyrophilin (BTN)-gene cluster, particularly the BTN3A2 gene, was observed by multilocus analyses. We replicated the associations with SNPs in the PRSS16 region and, albeit weaker, to the BTN3A2 region, in an independent material of 725 families obtained from the Type 1 Diabetes Genetics Consortium. It is important to note that these associations were independent of the HLA-DRB1-DQA1-DQB1 genes, as well as of associations observed at HLA-A, -B and -C. Taken together, our results identify PRSS16 and BTN3A2, two genes thought to play important roles in regulating the immune response, as potentially novel susceptibility genes for T1D. PMID- 19295543 TI - Genetic association of HLA DQB1 with CD4+CD25+(high) T-cell apoptosis in type 1 diabetes. AB - Type 1 diabetes (T1D) has a strong genetic component and the major locus lies in the HLA DQB1 region. We found earlier an increased apoptosis with decreased viability and function of the CD4+CD25+(high) T-cell subset (Treg) in human subjects with recent-onset T1D and in multiple autoantibody-positive, high at risk individuals. Tregs normally inhibit or delay onset of T1D in animal models and increased Treg apoptosis could bring on or accelerate disease from effector T cell-mediated destruction of insulin-producing beta cells. In this study, we test the hypothesis that HLA DQB1 genotypes are associated with increased CD4+CD25+(high) T-cell apoptosis. HLA DQ-based genetic risk status was significantly associated with CD4+CD25+(high) T-cell apoptosis, after adjustment for age, gender and phenotypic status (n=83, F=4.04 (d.f.=3), P=0.01). Unaffected, autoantibody-negative high risk HLA DQB1 control subjects showed increased CD4+CD25+(high) apoptosis levels compared with low risk HLA DQB1 control subjects (n=26, P=0.002), confirming that the association precedes disease. The association of specific HLA DQB1 genotypes with Treg apoptosis was also tested, showing significance for HLA DQB1*0302, DQB1*0201 and HLA DQB1*0602 alleles. Our study shows an association of HLA DQB1 genotypes with CD4+CD25+(high) T-cell apoptosis, which implicates CD4+CD25+(high) T-cell apoptosis as a new intermediate trait for T1D. PMID- 19295544 TI - The complete evaluation of erythrocytosis: congenital and acquired. AB - The approach to a patient with erythrocytosis is greatly simplified by assessing the clonality of the process upfront. In this regard, there has been a dramatic shift toward genetic testing and away from traditional tests, such as measurement of red cell mass. Clonal erythrocytosis is the diagnostic feature of polycythemia vera (PV) and is almost always associated with a JAK2 mutation (JAK2V617F or exon 12). All other scenarios represent non-clonal erythrocytosis, often referred to as secondary erythrocytosis. Serum erythropoietin (Epo) level is usually normal or elevated in secondary erythrocytosis and subnormal in PV. Therefore, in a patient with acquired erythrocytosis, it is reasonable to begin the diagnostic work-up with peripheral blood JAK2 mutation analysis and serum Epo measurement to distinguish PV from secondary erythrocytosis. Conversely, the patient with life long erythrocytosis is more likely to suffer from congenital polycythemia and should therefore be evaluated for germline mutations that result in enhanced Epo effect (for example, Epo receptor mutations), altered intracellular oxygen sensing (for example, mutations involving the von Hippel-Lindau tumor suppressor gene) or decreased P50 (for example, high-oxygen-affinity hemoglobinopathy). The order of tests in this instance depends on the clinical scenario and serum Epo level. PMID- 19295545 TI - Clonal expansion of T/NK-cells during tyrosine kinase inhibitor dasatinib therapy. AB - Dasatinib, a broad-spectrum tyrosine kinase inhibitor (TKI), predominantly targets BCR-ABL and SRC oncoproteins and also inhibits off-target kinases, which may result in unexpected drug responses. We identified 22 patients with marked lymphoproliferation in blood while on dasatinib therapy. Clonality and immunophenotype were analyzed and related clinical information was collected. An abrupt lymphocytosis (peak count range 4-20 x 10(9)/l) with large granular lymphocyte (LGL) morphology was observed after a median of 3 months from the start of therapy and it persisted throughout the therapy. Fifteen patients had a cytotoxic T-cell and seven patients had an NK-cell phenotype. All T-cell expansions were clonal. Adverse effects, such as colitis and pleuritis, were common (18 of 22 patients) and were preceded by LGL lymphocytosis. Accumulation of identical cytotoxic T cells was also detected in pleural effusion and colon biopsy samples. Responses to dasatinib were good and included complete, unexpectedly long-lasting remissions in patients with advanced leukemia. In a phase II clinical study on 46 Philadelphia chromosome-positive acute lymphoblastic leukemia, patients with lymphocytosis had superior survival compared with patients without lymphocytosis. By inhibiting immunoregulatory kinases, dasatinib may induce a reversible state of aberrant immune reactivity associated with good clinical responses and a distinct adverse effect profile. PMID- 19295546 TI - CYT387, a selective JAK1/JAK2 inhibitor: in vitro assessment of kinase selectivity and preclinical studies using cell lines and primary cells from polycythemia vera patients. AB - Somatic mutations in Janus kinase 2 (JAK2), including JAK2V617F, result in dysregulated JAK-signal transducer and activator transcription (STAT) signaling, which is implicated in myeloproliferative neoplasm (MPN) pathogenesis. CYT387 is an ATP-competitive small molecule that potently inhibits JAK1/JAK2 kinases (IC(50)=11 and 18 nM, respectively), with significantly less activity against other kinases, including JAK3 (IC(50)=155 nM). CYT387 inhibits growth of Ba/F3 JAK2V617F and human erythroleukemia (HEL) cells (IC(50) approximately 1500 nM) or Ba/F3-MPLW515L cells (IC(50)=200 nM), but has considerably less activity against BCR-ABL harboring K562 cells (IC=58 000 nM). Cell lines harboring mutated JAK2 alleles (CHRF-288-11 or Ba/F3-TEL-JAK2) were inhibited more potently than the corresponding pair harboring mutated JAK3 alleles (CMK or Ba/F3-TEL-JAK3), and STAT-5 phosphorylation was inhibited in HEL cells with an IC(50)=400 nM. Furthermore, CYT387 selectively suppressed the in vitro growth of erythroid colonies harboring JAK2V617F from polycythemia vera (PV) patients, an effect that was attenuated by exogenous erythropoietin. Overall, our data indicate that the JAK1/JAK2 selective inhibitor CYT387 has potential for efficacious treatment of MPN harboring mutated JAK2 and MPL alleles. PMID- 19295547 TI - Translocation t(14;18) is not associated with inferior outcome in chronic lymphocytic leukemia. PMID- 19295548 TI - Bcr-Abl-mediated redox regulation of the PI3K/AKT pathway. AB - Bcr-Abl causes chronic myelogenous leukemia, a myeloproliferative disorder characterized by clonal expansion of hematopoietic progenitor cells. In this study, inducible expression of Bcr-Abl in TonB.210 cells is associated with increased production of intracellular reactive oxygen species (ROS), which is thought to play a role in survival signaling when generated at specific levels. Elevated ROS in Bcr-Abl-expressing cells were found to activate PI3k/Akt pathway members such as Akt and GSK3beta as well as downstream targets beta-catenin and Mcl-1. The activation of these proteins was inhibited by the flavoprotein inhibitor diphenyleneiodonium, which is commonly used to inhibit NADPH oxidase (Nox). This indicated that increased ROS might be related to increased activity of one member of the Nox family. Knock-down experiments using siRNA suggest that Nox-4 is the main source of increased ROS following Bcr-Abl expression. We showed that Bcr-Abl-induced ROS could also increase survival pathway signaling through redox inhibition of PP1alpha, a serine threonine phosphatase that negatively regulates the PI3k/Akt pathway. Overall our results demonstrate that Bcr-Abl expression increases Nox-4-generated ROS, which in turn increases survival signaling through PI3k/Akt pathway by inhibition of PP1alpha, thus contributing to the high level of resistance to apoptosis seen in these Bcr-Abl-expressing cells. PMID- 19295549 TI - Detection of mutant TET2 in myeloid malignancies other than myeloproliferative neoplasms: CMML, MDS, MDS/MPN and AML. PMID- 19295550 TI - Hippopotamus and whale phylogeny. AB - Thewissen et al. describe new fossils from India that apparently support a phylogeny that places Cetacea (that is, whales, dolphins, porpoises) as the sister group to the extinct family Raoellidae, and Hippopotamidae as more closely related to pigs and peccaries (that is, Suina) than to cetaceans. However, our reanalysis of a modified version of the data set they used differs in retaining molecular characters and demonstrates that Hippopotamidae is the closest extant family to Cetacea and that raoellids are the closest extinct group, consistent with previous phylogenetic studies. This topology supports the view that the aquatic adaptations in hippopotamids and cetaceans are inherited from their common ancestor. PMID- 19295554 TI - Health highway. PMID- 19295555 TI - Turkey censors evolution. PMID- 19295556 TI - Filling the void. PMID- 19295563 TI - Journal club. A geneticist views two theories of X-chromosome inactivation in a broad context. PMID- 19295569 TI - Society sues journal over right to reply. PMID- 19295570 TI - Europe's green billions. PMID- 19295572 TI - America's long hard road to climate-change law. PMID- 19295571 TI - Scientists in bone battle. PMID- 19295573 TI - Big interest in heavy drugs. PMID- 19295574 TI - Roche vows to keep Genentech culture. PMID- 19295575 TI - Incoming chief to tackle woes of US food and drug agency. PMID- 19295582 TI - Science journalism: Supplanting the old media? PMID- 19295583 TI - Clinical epidemiology: Archived answers. PMID- 19295584 TI - Backlogged system in Australia shuts out new investigators. PMID- 19295585 TI - We need more insight into what's worth paying for. PMID- 19295586 TI - Grant-writing offices would let scientists get on with research. PMID- 19295587 TI - Diversity of funding sources and topics is key to survival. PMID- 19295588 TI - Do nations go to war over water? PMID- 19295594 TI - Spectroscopy: Handedness in quick time. PMID- 19295595 TI - Cancer: The nuances of therapy. PMID- 19295597 TI - Geophysics: Hot blanket in Earth's deep crust. PMID- 19295598 TI - Dinosaurs: Fuzzy origins for feathers. PMID- 19295599 TI - Chemistry: Thinking outside the flask. PMID- 19295600 TI - Global change: West-side story of Antarctic ice. PMID- 19295601 TI - Neuroscience: Secret of synapse specificity. PMID- 19295602 TI - Activation of CaMKII in single dendritic spines during long-term potentiation. AB - Calcium/calmodulin-dependent kinase II (CaMKII) plays a central part in long-term potentiation (LTP), which underlies some forms of learning and memory. Here we monitored the spatiotemporal dynamics of CaMKII activation in individual dendritic spines during LTP using two-photon fluorescence lifetime imaging microscopy, in combination with two-photon glutamate uncaging. Induction of LTP and associated spine enlargement in single spines triggered transient ( approximately 1 min) CaMKII activation restricted to the stimulated spines. CaMKII in spines was specifically activated by NMDA receptors and L-type voltage sensitive calcium channels, presumably by nanodomain Ca(2+) near the channels, in response to glutamate uncaging and depolarization, respectively. The high degree of compartmentalization and channel specificity of CaMKII signalling allow stimuli-specific spatiotemporal patterns of CaMKII signalling and may be important for synapse-specificity of synaptic plasticity. PMID- 19295603 TI - Design and engineering of an O(2) transport protein. AB - The principles of natural protein engineering are obscured by overlapping functions and complexity accumulated through natural selection and evolution. Completely artificial proteins offer a clean slate on which to define and test these protein engineering principles, while recreating and extending natural functions. Here we introduce this method with the design of an oxygen transport protein, akin to human neuroglobin. Beginning with a simple and unnatural helix forming sequence with just three different amino acids, we assembled a four-helix bundle, positioned histidines to bis-histidine ligate haems, and exploited helical rotation and glutamate burial on haem binding to introduce distal histidine strain and facilitate O(2) binding. For stable oxygen binding without haem oxidation, water is excluded by simple packing of the protein interior and loops that reduce helical-interface mobility. O(2) affinities and exchange timescales match natural globins with distal histidines, with the remarkable exception that O(2) binds tighter than CO. PMID- 19295604 TI - Femtosecond characterization of vibrational optical activity of chiral molecules. AB - Optical activity is the result of chiral molecules interacting differently with left versus right circularly polarized light. Because of this intrinsic link to molecular structure, the determination of optical activity through circular dichroism (CD) spectroscopy has long served as a routine method for obtaining structural information about chemical and biological systems in condensed phases. A recent development is time-resolved CD spectroscopy, which can in principle map the structural changes associated with biomolecular function and thus lead to mechanistic insights into fundamental biological processes. But implementing time resolved CD measurements is experimentally challenging because CD is a notoriously weak effect (a factor of 10(-4)-10(-6) smaller than absorption). In fact, this problem has so far prevented time-resolved vibrational CD experiments. Here we show that vibrational CD spectroscopy with femtosecond time resolution can be realized when using heterodyned spectral interferometry to detect the phase and amplitude of the infrared optical activity free-induction-decay field in time (much like in a pulsed NMR experiment). We show that we can detect extremely weak signals in the presence of large achiral background contributions, by simultaneously measuring with a femtosecond laser pulse the vibrational CD and optical rotatory dispersion spectra of dissolved chiral limonene molecules. We have so far only targeted molecules in equilibrium, but it would be straightforward to extend the method for the observation of ultrafast structural changes such as those occurring during protein folding or asymmetric chemical reactions. That is, we should now be in a position to produce 'molecular motion pictures' of fundamental molecular processes from a chiral perspective. PMID- 19295605 TI - Hybrid organic-inorganic rotaxanes and molecular shuttles. AB - The tetravalency of carbon and its ability to form covalent bonds with itself and other elements enables large organic molecules with complex structures, functions and dynamics to be constructed. The varied electronic configurations and bonding patterns of inorganic elements, on the other hand, can impart diverse electronic, magnetic, catalytic and other useful properties to molecular-level structures. Some hybrid organic-inorganic materials that combine features of both chemistries have been developed, most notably metal-organic frameworks, dense and extended organic-inorganic frameworks and coordination polymers. Metal ions have also been incorporated into molecules that contain interlocked subunits, such as rotaxanes and catenanes, and structures in which many inorganic clusters encircle polymer chains have been described. Here we report the synthesis of a series of discrete rotaxane molecules in which inorganic and organic structural units are linked together mechanically at the molecular level. Structural units (dialkyammonium groups) in dumb-bell-shaped organic molecules template the assembly of essentially inorganic 'rings' about 'axles' to form rotaxanes consisting of various numbers of rings and axles. One of the rotaxanes behaves as a 'molecular shuttle': the ring moves between two binding sites on the axle in a large amplitude motion typical of some synthetic molecular machine systems. The architecture of the rotaxanes ensures that the electronic, magnetic and paramagnetic characteristics of the inorganic rings-properties that could make them suitable as qubits for quantum computers-can influence, and potentially be influenced by, the organic portion of the molecule. PMID- 19295606 TI - Temperature-dependent thermal diffusivity of the Earth's crust and implications for magmatism. AB - The thermal evolution of planetary crust and lithosphere is largely governed by the rate of heat transfer by conduction. The governing physical properties are thermal diffusivity (kappa) and conductivity (k = kapparhoC(P)), where rho denotes density and C(P) denotes specific heat capacity at constant pressure. Although for crustal rocks both kappa and k decrease above ambient temperature, most thermal models of the Earth's lithosphere assume constant values for kappa ( approximately 1 mm(2) s(-1)) and/or k ( approximately 3 to 5 W m(-1) K(-1)) owing to the large experimental uncertainties associated with conventional contact methods at high temperatures. Recent advances in laser-flash analysis permit accurate (+/-2 per cent) measurements on minerals and rocks to geologically relevant temperatures. Here we provide data from laser-flash analysis for three different crustal rock types, showing that kappa strongly decreases from 1.5-2.5 mm(2) s(-1) at ambient conditions, approaching 0.5 mm(2) s(-1) at mid-crustal temperatures. The latter value is approximately half that commonly assumed, and hot middle to lower crust is therefore a much more effective thermal insulator than previously thought. Above the quartz alpha-beta phase transition, crustal kappa is nearly independent of temperature, and similar to that of mantle materials. Calculated values of k indicate that its negative dependence on temperature is smaller than that of kappa, owing to the increase of C(P) with increasing temperature, but k also diminishes by 50 per cent from the surface to the quartz alpha-beta transition. We present models of lithospheric thermal evolution during continental collision and demonstrate that the temperature dependence of kappa and C(P) leads to positive feedback between strain heating in shear zones and more efficient thermal insulation, removing the requirement for unusually high radiogenic heat production to achieve crustal melting temperatures. Positive feedback between heating, increased thermal insulation and partial melting is predicted to occur in many tectonic settings, and in both the crust and the mantle, facilitating crustal reworking and planetary differentiation. PMID- 19295607 TI - Obliquity-paced Pliocene West Antarctic ice sheet oscillations. AB - Thirty years after oxygen isotope records from microfossils deposited in ocean sediments confirmed the hypothesis that variations in the Earth's orbital geometry control the ice ages, fundamental questions remain over the response of the Antarctic ice sheets to orbital cycles. Furthermore, an understanding of the behaviour of the marine-based West Antarctic ice sheet (WAIS) during the 'warmer than-present' early-Pliocene epoch ( approximately 5-3 Myr ago) is needed to better constrain the possible range of ice-sheet behaviour in the context of future global warming. Here we present a marine glacial record from the upper 600 m of the AND-1B sediment core recovered from beneath the northwest part of the Ross ice shelf by the ANDRILL programme and demonstrate well-dated, approximately 40-kyr cyclic variations in ice-sheet extent linked to cycles in insolation influenced by changes in the Earth's axial tilt (obliquity) during the Pliocene. Our data provide direct evidence for orbitally induced oscillations in the WAIS, which periodically collapsed, resulting in a switch from grounded ice, or ice shelves, to open waters in the Ross embayment when planetary temperatures were up to approximately 3 degrees C warmer than today and atmospheric CO(2) concentration was as high as approximately 400 p.p.m.v. (refs 5, 6). The evidence is consistent with a new ice-sheet/ice-shelf model that simulates fluctuations in Antarctic ice volume of up to +7 m in equivalent sea level associated with the loss of the WAIS and up to +3 m in equivalent sea level from the East Antarctic ice sheet, in response to ocean-induced melting paced by obliquity. During interglacial times, diatomaceous sediments indicate high surface-water productivity, minimal summer sea ice and air temperatures above freezing, suggesting an additional influence of surface melt under conditions of elevated CO(2). PMID- 19295608 TI - Modelling West Antarctic ice sheet growth and collapse through the past five million years. AB - The West Antarctic ice sheet (WAIS), with ice volume equivalent to approximately 5 m of sea level, has long been considered capable of past and future catastrophic collapse. Today, the ice sheet is fringed by vulnerable floating ice shelves that buttress the fast flow of inland ice streams. Grounding lines are several hundred metres below sea level and the bed deepens upstream, raising the prospect of runaway retreat. Projections of future WAIS behaviour have been hampered by limited understanding of past variations and their underlying forcing mechanisms. Its variation since the Last Glacial Maximum is best known, with grounding lines advancing to the continental-shelf edges around approximately 15 kyr ago before retreating to near-modern locations by approximately 3 kyr ago. Prior collapses during the warmth of the early Pliocene epoch and some Pleistocene interglacials have been suggested indirectly from records of sea level and deep-sea-core isotopes, and by the discovery of open-ocean diatoms in subglacial sediments. Until now, however, little direct evidence of such behaviour has been available. Here we use a combined ice sheet/ice shelf model capable of high-resolution nesting with a new treatment of grounding-line dynamics and ice-shelf buttressing to simulate Antarctic ice sheet variations over the past five million years. Modelled WAIS variations range from full glacial extents with grounding lines near the continental shelf break, intermediate states similar to modern, and brief but dramatic retreats, leaving only small, isolated ice caps on West Antarctic islands. Transitions between glacial, intermediate and collapsed states are relatively rapid, taking one to several thousand years. Our simulation is in good agreement with a new sediment record (ANDRILL AND-1B) recovered from the western Ross Sea, indicating a long term trend from more frequently collapsed to more glaciated states, dominant 40 kyr cyclicity in the Pliocene, and major retreats at marine isotope stage 31 ( approximately 1.07 Myr ago) and other super-interglacials. PMID- 19295609 TI - An Early Cretaceous heterodontosaurid dinosaur with filamentous integumentary structures. AB - Ornithischia is one of the two major groups of dinosaurs, with heterodontosauridae as one of its major clades. Heterodontosauridae is characterized by small, gracile bodies and a problematic phylogenetic position. Recent phylogenetic work indicates that it represents the most basal group of all well-known ornithischians. Previous heterodontosaurid records are mainly from the Early Jurassic period (205-190 million years ago) of Africa. Here we report a new heterodontosaurid, Tianyulong confuciusi gen. et sp. nov., from the Early Cretaceous period (144-99 million years ago) of western Liaoning Province, China. Tianyulong extends the geographical distribution of heterodontosaurids to Asia and confirms the clade's previously questionable temporal range extension into the Early Cretaceous period. More surprisingly, Tianyulong bears long, singular and unbranched filamentous integumentary (outer skin) structures. This represents the first confirmed report, to our knowledge, of filamentous integumentary structures in an ornithischian dinosaur. PMID- 19295610 TI - Defensin-like polypeptide LUREs are pollen tube attractants secreted from synergid cells. AB - For more than 140 years, pollen tube guidance in flowering plants has been thought to be mediated by chemoattractants derived from target ovules. However, there has been no convincing evidence of any particular molecule being the true attractant that actually controls the navigation of pollen tubes towards ovules. Emerging data indicate that two synergid cells on the side of the egg cell emit a diffusible, species-specific signal to attract the pollen tube at the last step of pollen tube guidance. Here we report that secreted, cysteine-rich polypeptides (CRPs) in a subgroup of defensin-like proteins are attractants derived from the synergid cells. We isolated synergid cells of Torenia fournieri, a unique plant with a protruding embryo sac, to identify transcripts encoding secreted proteins as candidate molecules for the chemoattractant(s). We found two CRPs, abundantly and predominantly expressed in the synergid cell, which are secreted to the surface of the egg apparatus. Moreover, they showed activity in vitro to attract competent pollen tubes of their own species and were named as LUREs. Injection of morpholino antisense oligomers against the LUREs impaired pollen tube attraction, supporting the finding that LUREs are the attractants derived from the synergid cells of T. fournieri. PMID- 19295612 TI - Improved cutaneous healing in diabetic mice exposed to healthy peripheral circulation. AB - Impaired repair of skin defects is a major complication of diabetes; yet, the pathophysiology of diabetic (db) wound healing remains largely opaque. Here, we investigate the role of humoral factors in modulating db wound repair by generating chimeric animals through parabiotic joining of wild-type (wt) and diabetic (db/db) mice. This strategy allows wounds on healing-deficient db/db mice to be exposed to factors derived from the wt circulation at physiologically appropriate concentrations. When compared with db controls, chimeric db/db animals showed significantly improved healing of full-thickness, cutaneous wounds, with enhanced granulation tissue formation, angiogenesis, cell proliferation, and collagen deposition. Glycemic control was unaffected by parabiosis; however, the distribution of circulating leukocytes, altered in db controls, normalized in db-chimeras. Both wt and db cells were recruited from circulation into db wounds, but wt cells never exceeded 20% of total cells. Improved angiogenesis persisted in db-chimeras separated 24 hours after wounding, suggesting the existence of long-term normalizing factors. This study establishes a new model for studying db wound healing, and shows a key role for circulating factors in normalizing wound repair in diabetes. PMID- 19295613 TI - Distinguishing mouse strains by proteomic analysis of pelage hair. AB - AKR/J mice display a hair interior defect (hid) phenotype for which the molecular basis is unknown. To investigate the application of hair-shaft proteomics to the study of such diseases, pelage from AKR/J and two other mouse strains without this defect was analyzed by shotgun proteomics. The results permitted the identification of 111 proteins from tryptic digests of total hair from AKR/J hid/hid mice, which were predominantly keratins (Krts) and Krt-associated proteins (Krtaps). From the non-solubilizable (crosslinked) fraction of the hair remaining after extensive detergent extraction, 58 proteins were identified. The majority were Krts and Krtaps, but junctional and other membrane proteins, cytoplasmic proteins, and histones were also identified. The results indicate the incorporation of a multitude of proteins into highly crosslinked material. Comparison of unique peptides generated among hair samples from AKR/J-hid/hid, FVB/NJ+/+, and LP/J+/+ mice indicated that these inbred strains could be distinguished by their proteomic patterns. Transmission electron microscopy after mild treatment in detergent and reducing agent permitted the visualization of projections of cortex cells, with characteristic filament patterns, into adjoining medulla cells. Hair shafts from AKR/J mice were deficient in these projections and also exhibited relatively low levels of trichohyalin, a possible contributor to or marker for the hid phenotype. PMID- 19295614 TI - Th17 cytokines stimulate CCL20 expression in keratinocytes in vitro and in vivo: implications for psoriasis pathogenesis. AB - T helper (Th) 17 cells have recently been implicated in psoriasis pathogenesis, but mechanisms of how these cells traffic into inflamed skin are unknown. By immunostaining for interleukin (IL)-17A and IL-22, we show numerous cells present in psoriasis lesions that produce these cytokines. We next found that Th17 cytokines (IL-17A, IL-22, and tumor necrosis factor (TNF)-alpha) markedly increased the expression of CC chemokine ligand (CCL) 20, a CC chemokine receptor (CCR)6 ligand, in human keratinocyte monolayer and raft cultures in a dose- and time-dependent manner. Lastly, we showed in mice that subcutaneous injection with recombinant IL-17A, IL-22, or TNF-alpha led to the upregulation of both CCL20 and CCR6 expression in skin as well as cutaneous T-cell infiltration. Taken together, these data show that Th17 cytokines stimulate CCL20 production in vitro and in vivo, and thus provide a potential explanation of how CCR6-positive Th17 cells maintain their continual presence in psoriasis through a positive chemotactic feedback loop. PMID- 19295615 TI - N-propionyl-cysteaminylphenol-magnetite conjugate (NPrCAP/M) is a nanoparticle for the targeted growth suppression of melanoma cells. AB - A magnetite nanoparticle, NPrCAP/M, was produced for intracellular hyperthermia treatment of melanoma by conjugating N-propionyl-cysteaminylphenol (NPrCAP) with magnetite and used for the study of selective targeting and degradation of melanoma cells. NPrCAP/M, like NPrCAP, was integrated as a substrate in the oxidative reaction by mushroom tyrosinase. Melanoma, but not non-melanoma, cells incorporated larger amounts of iron than magnetite from NPrCAP/M. When mice bearing a B16F1 melanoma and a lymphoma on opposite flanks were given NPrCAP/M, iron was observed only in B16F1 melanoma cells and iron particles (NPrCAP/M) were identified within late-stage melanosomes by electron microscopy. When cells were treated with NPrCAP/M or magnetite and heated to 43 degrees C by an external alternating magnetic field (AMF), melanoma cells were degraded 1.7- to 5.4-fold more significantly by NPrCAP/M than by magnetite. Growth of transplanted B16 melanoma was suppressed effectively by NPrCAP/M-mediated hyperthermia, suggesting a clinical application of NPrCAP/M to lesional therapy for melanoma. Finally, melanoma cells treated with NPrCAP/M plus AMF showed little sub-G1 fraction and no caspase 3 activation, suggesting that the NPrCAP/M-mediated hyperthermia induced non-apoptotic cell death. These results suggest that NPrCAP/M may be useful in targeted therapy for melanoma by inducing non-apoptotic cell death after appropriate heating by the AMF. PMID- 19295616 TI - Differential effects of corticosteroids and pimecrolimus on the developing skin immune system in humans and mice. AB - Atopic dermatitis arises primarily in early infancy. In these patients, corticosteroids are used especially with great caution because of their side effects. Calcineurin inhibitors such as pimecrolimus (PIM) could be useful, but safety concerns have been raised in particular because of the lack of knowledge about their effects on the developing skin immune system. This study was designed to investigate the impact of PIM and corticosteroids on epidermal cells (EC) in infants and newborn mice. We found that the percentage of unfractionated viable infant ECs was significantly decreased in the presence of beta-methasone-17 valerate (BMV) but not PIM. Exposure of unfractionated infant ECs to BMV but not to PIM and vehicle control caused a significant inhibition of the upregulation of CD86 molecules on Langerhans cells (LC). The release of cytokines by LCs and ECs, cultured in the presence of BMV and PIM, was not significantly reduced compared with controls. Topical corticosteroid but not PIM application onto newborn mice induced apoptosis in some LC precursors. Our data suggest that similar to the situation in adult skin, corticosteroids may impair LC maturation as well as viability of ECs in infants, effects not seen with PIM. PMID- 19295617 TI - Identity in the contemporary world. PMID- 19295618 TI - Contemporary impingements on mothering. AB - Mothering in contemporary Western society needs to be understood in the context of a rapidly changing social context. Increased geographic mobility, improved access to child-related information through the media, and scientific and technological progress have contributed to significant shifts in cultural views on mothering. Several contextual impingements on mothering, including changing family structure, economic pressures, decreased social support, cultural ideals of the perfect mother, and increased awareness of interpersonal and global trauma impact mothers' internal worlds. These societal changes often reinforce mothers' fear of losing their children and an idealization of intensive mothering, and evoke challenges in reorganizing their sense of personal identity. Implications for psychoanalytic theory and practice, and specifically the need to integrate individual and contextual forces related to experiences of mothers will be explored. PMID- 19295619 TI - Who are we? Where did we come from? How religious identity divides and damns us all. AB - Charles Darwin's theory of natural selection, now established, is the only workable explanation we have for the remarkable fact of our own existence, the vast diversity of plant and animal life, the compelling illusion of design in nature, and the architecture of the human mind. We are risen apes, not fallen angels. Our different races and ethnicities hide an essential truth: we are all Africans, all siblings, all descendants of a small group of hunter-gatherers who arose in Africa less than 200,000 years ago and conquered the world. There is one universal human nature stemming from our long history as Pleistocene hunter gatherers. We now know why religion appeared, why particular religious ideas emerged, why they are widespread, why they are recurrent features of human minds and human societies, why they are attractive to human minds, and how and why they are related to survival and deadly violence. What makes us human can also make us religious. Religion is a by-product of cognitive mechanisms that evolved, through natural selection, for other adaptive purposes that were crucial for our survival and reproductive success. Religion's power derives from its ability to utilize these mental mechanisms designed for other tasks: our attachment to parental figures, our wish for help in distress, submission to authority, our sensitivity to hierarchies, our love of kin, and our inclination to see human-like intention as the explanation for any unknown. All individuals are vulnerable to religious beliefs and assuming a religious identity. Religious identity is an accident of geographical birth, is certainly one of the most prevalent identities, and the most at odds with the reality of who we are. It remains one of the most destructive identities we can assume. As long as we tolerate the divisive pretensions of religious beliefs and identities, especially those of the three great Abrahamic faiths, we will all remain in the line of fire. PMID- 19295620 TI - Identity transformation in medical students. AB - This paper reviews the literature on the impact of medical school on personal development and consolidation of core identity. The limited literature relies on reports from medical students' journaling exercises, discussion groups, post graduation surveys, and repeated personality testing. We review forces acting on medical students, with potential transforming effects. These forces include high external expectations and internal fear of superficial knowledge and skills, entry into the culture of medicine with its insider jargon and hierarchy, high academic workload, and the emotional burdens of confronting cadavers and death as well as bearing witness to patients' suffering. Potential developmental delay, emergence of substance abuse and hedonic acting out, cynicism, and loss of individual core values are possible consequences. Protections against these adverse outcomes include identification of strong mentors and role models, developing post-conventional morality and relativistic thinking, finding healthy coping strategies such as peer support, and remaining intellectually creative and personally reflective. PMID- 19295621 TI - The Identity Consolidation Inventory (ICI): development and application of a questionnaire for assessing the structuralization of individual identity. AB - This paper describes the author's effort to develop a self-rating scale to measure the consolidation of identity in a given individual. It describes the construction of the 35-item Identity Consolidation Inventory (ICI) and, as an appendix to the paper, includes the final version of this questionnaire. The scale's reliability and validity are demonstrated via its administration to 238 normal controls, 80 individuals with Borderline Personality Disorder and 13 subjects with Dissociative Identity Disorder ("multiple personality"). The clinical and research usefulness of ICI lies in the realm of discerning the syndrome of identity diffusion that underlies all severe personality disorders. PMID- 19295622 TI - The Palestinian/Israeli Conflict: a geopolitical identity disorder. AB - In this paper, the author considers that the large-group dynamics in certain war torn, hot spots throughout the world are symptoms of a "geopolitical identity disorder." He extrapolates from the model of the severely traumatized psyche in dissociative identity disorder in which there is so much intolerable emotion, destructive aggression and conflict that different selves with different identities develop which are unable to recognize how interdependent and related they actually are. In the most extreme cases, one dissociated self tries to kill off "the other" in order to gain exclusive control of the body and make it comply with his or her wishes and vision. In actuality, however, such an attempt is a dissociated suicide plan with lethal implications. This model is applied to the Palestinian/Israeli situation where there is a deadly battle over the land. A clinical vignette is offered to illustrate these ideas and offer possibilities for help. PMID- 19295627 TI - Embrace the complexity. PMID- 19295623 TI - State-dependent self-representations: a culture-bound aspect of identity. AB - The concepts of identity, self and self-representation have been discussed extensively in psychoanalytic metapsychology. These concepts are at times confusing and are used interchangeably by various authors. Regardless of what one calls it, what one experiences in a given moment is one's representation as an analyst or a father or a son or daughter, depending on the situation one is in. This paper describes such state-dependent self-representations as an aspect of the self and argues that state-dependent self-representations are probably more clinically relevant and useful in day-to-day practice. PMID- 19295628 TI - Navigating the network: signaling cross-talk in hematopoietic cells. AB - Recent studies in hematopoietic cells have led to a growing appreciation of the diverse modes of molecular and functional cross-talk between canonical signaling pathways. However, these intersections represent only the tip of the iceberg. Emerging global analytical methods are providing an even richer and more complete picture of the many components that measurably interact in a network manner to produce cellular responses. Here we highlight the pieces in this Focus, emphasize the limitations of the present canonical pathway paradigm, and discuss the value of a systems biology approach using more global, quantitative experimental design and data analysis strategies. Lastly, we urge caution about overly facile interpretation of genome- and proteome-level studies. PMID- 19295629 TI - Integration of cytokine and heterologous receptor signaling pathways. AB - Cytokines are soluble mediators of cell communication that are critical in immune regulation. They induce specific gene-expression programs in responsive cells. Recent findings, however, indicate that cytokine receptors can regulate immune cell functions by transcription-independent mechanisms. Here we review the current understanding of how cytokine signaling regulates the functions of other signaling pathways by first discussing the 'traditional' transcription-mediated consequences of cytokine signaling and then providing a detailed description of transcription-independent lateral communications between cytokine receptors and other cellular receptors. PMID- 19295630 TI - Cross-regulation of signaling by ITAM-associated receptors. AB - An important function of receptors that signal through immunoreceptor tyrosine based activation motifs (ITAMs) is to regulate signaling by heterologous receptors. This review describes mechanisms by which ITAM-associated receptors modulate signaling by Toll-like receptors (TLRs), tumor necrosis factor receptor family members and cytokine receptors that use the Jak-STAT signaling pathway, and the biological importance of this signal transduction cross-talk. ITAM mediated cross-regulation can either augment or dampen signaling by other receptors. Conversely, TLRs and cytokines modulate ITAM-mediated signaling, by means including activation of beta2 integrins that are coupled to the ITAM containing adaptors DAP12 and FcRgamma. Integration of ITAM signaling into signaling networks through cross-talk with other signal transduction pathways results in tight regulation and fine tuning of cellular responses to various extracellular stimuli and contributes to induction of specific activation and differentiation pathways. PMID- 19295631 TI - Death receptor signal transducers: nodes of coordination in immune signaling networks. AB - Death receptors (DRs) are members of the tumor necrosis factor receptor superfamily that possess a cytoplasmic death domain (DD). DRs regulate important operational and homeostatic aspects of the immune system. They transmit signals through apical protein complexes, which are nucleated by the DD adaptors FADD and TRADD, to control cellular outcomes that range from apoptosis to gene activation. FADD and TRADD also nucleate several distal signaling complexes, which mediate cross-talk between distinct DR signaling pathways. Moreover, together with other DR signal transducers, FADD and TRADD participate in functional complexes assembled by certain non-DR immune cell receptors, such as pattern-recognition receptors. Thus, DR signal transducers may provide important nodes of coordination in immune signaling networks. PMID- 19295632 TI - Selectivity and therapeutic inhibition of kinases: to be or not to be? AB - Protein kinases, which serve critical functions in signaling pathways in all cells, are popular therapeutic targets. At present, eight kinase inhibitors have been approved in the United States, each of which shows nanomolar potency. Although the initial goal was to generate inhibitors with a high degree of selectivity, recent experience has revealed that many of these approved compounds target more than one kinase. Surprisingly, this promiscuity is less problematic than one would have imagined; indeed, it opens new therapeutic opportunities. In this Perspective, we discuss the present status of Janus kinase inhibitors-a new class of immunosuppressive drugs-and the advantages and disadvantages of selectively inhibiting this class of kinase. PMID- 19295633 TI - Lymphocyte signaling: beyond knockouts. AB - The analysis of lymphocyte signaling was greatly enhanced by the advent of gene targeting, which allows the selective inactivation of a single gene. Although this gene 'knockout' approach is often informative, in many cases, the phenotype resulting from gene ablation might not provide a complete picture of the function of the corresponding protein. If a protein has multiple functions within a single or several signaling pathways, or stabilizes other proteins in a complex, the phenotypic consequences of a gene knockout may manifest as a combination of several different perturbations. In these cases, gene targeting to 'knock in' subtle point mutations might provide more accurate insight into protein function. However, to be informative, such mutations must be carefully based on structural and biophysical data. PMID- 19295635 TI - AID and RPA: PKA makes the connection local. PMID- 19295636 TI - The road not taken: memory T cell fate 'decisions'. PMID- 19295637 TI - Revisiting the follicular helper T cell paradigm. PMID- 19295639 TI - Pyrococcus CH1, an obligate piezophilic hyperthermophile: extending the upper pressure-temperature limits for life. AB - A novel hydrothermal site was discovered in March 2007, on the mid-Atlantic ridge during the cruise 'Serpentine'. At a depth of 4100 m, the site 'Ashadze' is the deepest vent field known so far. Smoker samples were collected with the ROV 'Victor 6000' and processed in the laboratory for the enrichment of anaerobic heterotrophic microorganisms under high-temperature and high-hydrostatic pressure conditions. Strain CH1 was successfully isolated and assigned to the genus Pyrococcus, within the Euryarchaeota lineage within the Archaea domain. This organism grows within a temperature range of 80 to 108 degrees C and a pressure range of 20 to 120 MPa, with optima for 98 degrees C and 52 MPa respectively. Pyrococcus CH1 represents the first obligate piezophilic hyperthermophilic microorganism known so far. Comparisons of growth yields obtained under high temperature/high-pressure conditions for relative organisms isolated from various depths, showed clear relationships between depth at origin and responses to hydrostatic pressure. PMID- 19295640 TI - Microbiomic analysis of the bifidobacterial population in the human distal gut. AB - One of the most complex microbial ecosystems is represented by the microbiota of the human gastrointestinal tract (GIT). Although this microbial consortium has been recognized to have a crucial effect on human health, its precise composition is still not fully established. Among the GIT bacteria, bifidobacteria represent an important commensal group whose presence is often associated with health promoting effects. In this work, we assessed the complexity of the human intestinal bifidobacterial population by analysing the diversity of several 16S rRNA gene-based libraries. These analyses showed the presence of novel bifidobacterial phylotypes, which had not been found earlier and may thus represent novel taxa within the genus Bifidobacterium. PMID- 19295641 TI - [Virus and prophages in aquatic ecosystems]. AB - In this review, available data on the structure (diversity, abundance, biomass) and functional imprints (bacteriolysis, lysogeny, gene transfers, regulation of prokaryotic diversity) of natural viruses in the context of food webs in aquatic microbial ecology, and the related biogeochemical cycles, are summarized. Viruses are the most abundant, and probably the most diverse, biological entities in aquatic ecosystems and in the biosphere (i.e., viriosphere). Aquatic viruses typically exceed 107 particles/mL in mesotrophic conditions, the majority being represented by phages without tails and by tailed-phages such as members of the family Siphoviridae. Both types of phages have a small capsid and a small genome size, which is considered an evolutionary adaptation to planktonic life. Their contribution to microbial mortality is significant. There is strong evidence that phages exert a significant pressure on the community structure and diversity and on the diversification of potential hosts, mainly through two major pathways: biogeochemical catalysis from lysis products and horizontal gene transfers. In turn, phages are sensitive to environmental factors, both in terms of integrity and of infectivity. Some phages contain typical viral genes that code for biological functions of interest, such as photosynthesis. In general, development in viral ecology is a source of new knowledge for the scientific community in the domain of environmental sciences, but also in the context of evolutionary biology of living cellular organisms, the obligatory hosts for viruses. For example, the recent discovery of a giant virus that becomes ill through infection by another virus (i.e., a viriophage) is fuelling debate about whether viruses are alive. Finally, future research directions are identified in the context of general aquatic ecology, including ecological researches on cyanophages and other phytoplanktonic phages as a priority, primarily in freshwater lakes. PMID- 19295642 TI - Extracellular superoxide anion production contributes to the virulence of Xanthomonas oryzae pv. oryzae. AB - Endogenous superoxide anion production was determined by electron spin resonance in wild-type strains and avrXa7 mutants of Xanthomonas oryzae pv. oryzae. The localization of superoxide anion was carried out in the intra- and extra-cellular fractions. Results showed the presence of superoxide anion in multi-locations of X. oryzae pv. oryzae cells. The extracellular fraction was the major location of superoxide anion production. Furthermore, a positive relationship was shown between the levels of endogenous superoxide anion and the virulence of strains. These indubitable results suggested that the superoxide anion contributes to the virulence of X. oryzae pv. oryzae. PMID- 19295643 TI - Whole-cell protein and ITS rDNA profiles as diagnostic tools to discriminate Fusarium avenaceum intraspecific variability and associated virulence. AB - A total of 91 isolates of Fusarium avenaceum were regrouped into 15 phenotypes and 10 vegetative compatibility groups showing specific one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (1-D SDS-PAGE) protein profiles and less-specific internal transcribed spacer rDNA profiles. Each isolate possessed reproducible signature protein bands. Indeed, the unweighted pair group method with arithmetic averages clustering revealed that the protein profile of each group of isolates correlated with fungus virulence. The use of SDS-PAGE offers a simple and sensitive technique for routine differentiation between pathogenic and nonpathogenic isolates within unknown F. avenaceum populations. The discovery has significant implications for risk assessment of cereal yield to ensure food and feed safety. This low-cost approach has the potential to be optimized and extended to a broad spectrum of Fusarium head blight pathogens. PMID- 19295644 TI - Helicobacter pylori detection and genotyping in gastric biopsy specimens from Chennai patients (India). AB - The aim of the present study was to investigate the prevalence of Helicobacter pylori infection and the correlation between cagA and vacA (mid-region) genotypes with different clinical outcomes from Chennai, India, patients. Biopsies from the antrum were taken to assess the current H. pylori status by histology, rapid urease test (RUT), and PCR. The RUT and PCR analyses were carried out on a single biopsy specimen. Fasting sera were obtained from all patients and H. pylori status was determined by using ELISA. In addition, the correlations between cagA and vacA genotypes and the consequence of H. pylori infection were statistically examined. Prevalence of the cagA gene was found in 96% (90/94) of patients, and the vacA m2 subtype occurred in 60% (56/94), whereas 32% (30/94) showed the vacA m1 subtype. A significant association between the cagA and vacA m2 region (chi2 = 5.556; p < 0.01) was found in ulcer patients. The vacA m2 genotype showed a near significant value (chi2 = 3.943; p < 0.047) in ulcer patients when compared with vacA m1. These findings suggest that H. pylori strains with the vacA m2 region were predominant in South India, especially in and around Chennai. This study also showed that PCR has a potential value for studying the cagA gene directly from biopsy specimens. PMID- 19295645 TI - Improved cure of bacterial vaginosis with single dose of tinidazole (2 g), Lactobacillus rhamnosus GR-1, and Lactobacillus reuteri RC-14: a randomized, double-blind, placebo-controlled trial. AB - Bacterial vaginosis (BV) is the most prevalent vaginal infection worldwide and is characterized by depletion of the indigenous lactobacilli. Antimicrobial therapy is often ineffective. We hypothesized that probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 might provide an adjunct to antimicrobial treatment and improve cure rates. Sixty-four Brazilian women diagnosed with BV were randomly assigned to receive a single dose of tinidazole (2 g) supplemented with either 2 placebo capsules or 2 capsules containing L. rhamnosus GR-1 and L. reuteri RC-14 every morning for the following 4 weeks. At the end of treatment (day 28), the probiotic group had a significantly higher cure rate of BV (87.5%) than the placebo group (50.0%) (p = 0.001). In addition, according to the Gram stain Nugent score, more women were assessed with "normal" vaginal microbiota in the probiotic group (75.0% vs. 34.4% in the placebo group; p = 0.011). This study shows that probiotic lactobacilli can provide benefits to women being treated with antibiotics for an infectious condition. PMID- 19295646 TI - Entry of dengue virus serotype 2 into ECV304 cells depends on clathrin-dependent endocytosis, but not on caveolae-dependent endocytosis. AB - Caveolae- and clathrin-mediated endocytosis are major internalization pathways used by several pathogens; however, their distinctive roles in dengue virus (DV) entry have not been addressed. In this study, we compared the involvement of caveolae- and clathrin-mediated endocytosis in the infectious entry of DV serotype 2 (DV2) into human endothelial-like ECV304 cells. Confocal microscopy study on DV2-infected cells showed that viral antigens were co-localized with clathrin heavy chains, epidermal growth factor pathway substrate clone 15 (Eps15), and adaptin-alpha, but not with caveolin-1. Treatment with chlorpromazine, which inhibits clathrin-dependent endocytosis, led to reduced virus entry into cells, whereas treatment with nystatin, a caveolae inhibitory agent, did not. Furthermore, gene silencing of Eps15 resulted in an average of 75% reduced infection of ECV304 cells by DV2. Our results demonstrated that DV2 enters ECV304 cells by clathrin-dependent endocytosis, not by caveolae-dependent endocytosis. PMID- 19295647 TI - A modified suspension test for estimating the mutagenicity of samples containing free and (or) protein-bound histidine. AB - The Ames test has not been very effective in estimating the mutagenicity of histidine-containing samples because external free and (or) protein-bound histidine in these samples would allow the histidine auxotrophs in such test samples to grow more compared with the negative controls that were used as the reference. This could give rise to a false positive.n this study, a modified suspension mutagenicity assay (MS assay) was developed. The tester strains were incubated in Luria-Bertani (LB) broth containing different concentrations of traditional Chinese medicines (TCMs) until the declining phase, and the test samples were assayed to be mutagenic or not by observing whether statistically significant differences were demonstrated in the relative reversion frequencies (RRFs) between the negative control groups and the test groups. Collectively, using LB broth as the test medium and comparing the RRFs in the declining phase made this assay less influenced by the presence of histidine in the test samples.The mutagenicity of some TCMs was measured with the MS assay. The results in MS assay were consistent with those in the mammalian bone marrow chromosomal aberration test, which indicated that the MS assay was appropriate to estimate the mutagenicity of samples containing free and (or) protein-bound histidine. PMID- 19295648 TI - Biodiversity of Trichoderma strains in Tunisia. AB - Trichoderma strains were sampled in 4 different bioclimatic zones of Tunisia, a Mediterranean North African country with strong climatic and edaphic variability from north to south, to assess the genetic diversity of endemic species of Trichoderma and their relationship to the bioclimatic zones. In all, 53 strains were isolated and identified at the species level by analysis of their internal transcribed spacers regions 1 and 2 (ITS1 and ITS2) of the rDNA cluster and (or) a fragment of the translation elongation factor 1 (tef1) gene, using an online interactive key for species identification in Trichoderma and ex-type strains and taxonomically established isolates of Trichoderma as references. At least 2 different species were observed in each ecosystem. Trichoderma harzianum clade VI and Trichoderma longibrachiatum were present in forest soils in north Tunisia; Trichoderma atroviride and Trichoderma hamatum were found in cultivated fields in northeast Tunisia; T. harzianum clade VI, a Trichoderma sp. close to the T. harzianum complex, and Trichoderma saturnisporum were isolated from forest soils in central Tunisia; and T. harzianum clade II and T. hamatum were present in oasis soils in south Tunisia. PMID- 19295649 TI - Morphological and biochemical changes in Pseudomonas fluorescens biofilms induced by sub-inhibitory exposure to antimicrobial agents. AB - Confocal laser scanning microscopy (CLSM) and scanning transmission X-ray microscopy (STXM) were used to examine the morphological and biochemical changes in Pseudomonas fluorescens biofilms grown in the presence of subinhibitory concentrations of 4 antimicrobial agents: triclosan, benzalkonium chloride, chlorhexidine dihydrochloride, and trisodium phosphate. CLSM analyses using the stains SYTO9 and propidium iodide indicated that the antimicrobial agents affected cell membrane integrity and cellular density to differing degrees. However, fluorescein diacetate assays and plate counts demonstrated that the cells remained metabolically active. Fluorescent lectin binding assays showed that changes in the arrangement and composition of the exopolymer matrix of the biofilms also occurred and that these changes depended on the antimicrobial agent. Detailed single cell analyses using STXM provided evidence that the cell morphology, and the spatial distribution and relative amounts of protein, lipids and polysaccharides in the biofilms and within the cells were different for each antimicrobial. The distribution of chlorhexidine in the biofilm, determined from its distinct spectral signature, was localized mainly inside the bacterial cells. Each antimicrobial agent elicited a unique response; P. fluorescens cells and biofilms changed their morphology and architecture, as well as the distribution and abundance of biomacromolecules, in particular the exopolymer matrix. Pseudomonas fluorescens also exhibited adaptation to benzalkonium chloride at 10 microg/mL. Our observations point to the importance of changes in the quantity and chemistry of the exopolymeric matrix in the response to antimicrobial agents and suggest their importance as targets for control. PMID- 19295650 TI - In vivo associations of Escherichia coli NarJ with a peptide of the first 50 residues of nitrate reductase catalytic subunit NarG. AB - The catalytic subunit of many Escherichia coli redox enzymes bares a twin arginine translocation (Tat)-dependent signal peptide in its precursor, which directs the redox enzyme complex to this Sec-independent pathway. NarG of the E. coli nitrate reductase NarGHI complex possesses a vestige twin-arginine motif at its N terminus. During the cofactor insertion, and assembly and folding of the NarG-NarH complex, a chaperone protein, NarJ, is thought to interact with the N terminus and an unknown second site of NarG. Our previous in vitro study provided evidence that NarJ's role shows some Tat system dependence. In this work, we investigated the associations of NarJ with a peptide of the first 50 residues of NarG (NarG50) in living cells. Two approaches were used: the Forster resonance energy transfer (FRET) based on yellow fluorescent protein - cyan fluorescent protein (YFP-CFP) and the bimolecular fluorescence complementation (BiFC). Compared with the wild-type (WT) E. coli cotransformants expressing both NarJ-YFP and NarG50-CFP, tat gene mutants gave an apparent FRET efficiency (Eapp) that was on the order of 25%-40% lower. These experiments implied a Tat system dependency of the in vivo associations between NarJ and the NarG50 peptide. In the BiFC assay, a 4-fold lower specific fluorescence intensity was observed for the E. coli WT cotransformants expressing both NarJ-Yc and NarG50-Yn than for its tat mutants, again suggesting a Tat dependence of the interactions. Fluorescence microscopy showed a "dot" /unipolar distribution of the reassembled YFP NarJ:NarG50 both in WT and tat mutants, demonstrating a distinct localization of the interaction. Thus, although the degree of the interaction shows Tat dependence, the cell localization is less so. Taken together, these data further support that NarJ's activity on NarG may be assisted by the Tat system. PMID- 19295651 TI - The impact of growth history and flagellation on the adhesion of various Listeria monocytogenes strains to polystyrene. AB - The contribution of growth history and flagella to adhesion of Listeria monocytogenes was analysed. An in-frame deletion on the flagellin encoding gene (flaA) was performed in L. monocytogenes EGD-e to compare its adhesion ability with the parental strain, after cultivation at various pH values and temperatures. The pH, as well as the temperature, affected the adhesion of L. monocytogenes EGD-e. In addition, the adhesion of L. monocytogenes EGD-e was reduced in energy-depressed cells. Conversely, the physicochemical bacterial surface characteristics affected by growth history did not influence the adhesion. Adhesion variations observed among environmental and clinical strains was attributed to the flagella. The naturally aflagellated strains resulted in an adhesion capacity similar to that observed for mutants and parental strains cultivated under flagellum expression repressing conditions. However, L. monocytogenes is able to adhere to inert surfaces through a residual adhesion process without flagella. All these observations emphasize the importance to consider the food environmental factors in the risk assessment of L. monocytogenes in food industry. PMID- 19295652 TI - Biofilm evidence and the microbial diversity of horse wounds. AB - Evidence of biofilms in human chronic wounds are thought to be responsible for preventing healing in a timely manner. However, biofilm evidence in horse wounds has not yet been documented. Consequently, this study aimed to determine whether biofilms could be detected in wounds, and to investigate the microbiology of chronic wounds in horses. Prior to analysis, wound surfaces were irrigated with 5 mL of sterile saline to remove debris. All wounds were swabbed twice (1 cm2 area) using sterile cotton-tipped swabs. In addition to this, 2 tissue biopsies were taken to investigate evidence of biofilm and the microbiology richness of the wounds. All swabs and 1 biopsy sample were transported to the laboratory in Robertson's cooked meat broth. Traditional culturable techniques and denaturing gradient gel electrophoresis with PCR were utilized to identify common bacteria isolated in all wounds. Following analysis of a number of the biopsy samples, biofilms could be clearly seen. The most common bacteria isolated from each wound analysed included Pseudomonas aeruginosa, Staphylococcus epidermidis, Serratia marcescens, Enterococcus faecalis, and Providencia rettgeri. Sequencing of the 16S ribosmonal DNAs, selected on the basis of DGGE profiling, enabled identification of bacterial species not identified using culturable technology. This study is the first to identify biofilms in the chronic wounds of horses. In addition, this study also demonstrated the importance of combining DGGE-PCR with culture techniques to provide better microbiology analysis of chronic wounds. PMID- 19295653 TI - Effect of biofumigation with volatiles from Muscodor albus on the viability of Tilletia spp. teliospores. AB - Volatile organic compounds produced by the fungus Muscodor albus inhibit or kill numerous fungi. The effect of these volatiles was tested on dormant and physiologically active teliospores of the smut fungi Tilletia horrida, Tilletia indica, and Tilletia tritici, which cause kernel smut of rice, Karnal bunt of wheat, and common bunt of wheat, respectively. Reactivated rye grain culture of M. albus was used to fumigate dormant teliospores in dry Petri dishes and physiologically active teliospores on water agar for up to 5 days at 22 degrees C. Teliospores of all 3 species were incapable of germination when fumigated on agar for 5 days. When T. tritici on agar was fumigated only during the initial 48 h of incubation, viability was reduced by 73%-99%. Fumigation of dry loose teliospores of T. tritici caused a 69%-97% loss in viability, whereas teliospores within intact sori were not affected. Dormant teliospores of T. horrida and T. indica were not affected by M. albus volatiles. It appears that M. albus has potential as a seed or soil treatment for controlling seedling-infecting smuts where infection is initiated by germinating teliospores prior to seedling emergence. The volatiles were not effective for postharvest control of teliospores under conditions used in these experiments. PMID- 19295654 TI - Detection of Bacillus sphaericus mosquitocidal toxin genes by multiplex colony PCR. AB - A multiplex colony PCR assay was developed for the detection of 5 genes encoding Bacillus sphaericus mosquito larvicidal toxins, namely binA, binB, mtx1, mtx2, and mtx3. Primers designed for these 5 genes yielded specific PCR amplicons of the expected size from type cultures of B. sphaericus. This method of detecting multiple toxin genes by colony PCR in a single tube reaction is a simple, rapid, and economical technique for identification of highly toxic environmental B. sphaericus isolates. PMID- 19295655 TI - Functional analysis of three AHL autoinducer synthase genes in Mesorhizobium loti reveals the important role of quorum sensing in symbiotic nodulation. AB - One of the most important signal transduction pathways in bacteria, quorum sensing, is involved in many regulatory circuits in rhizobia, especially in the control of communication between rhizobia and their plant hosts. In this study, we identified 3 autoinducer synthase genes - mrlI1, mrlI2, and mrlI3 - in Mesorhizobium loti NZP 2213. We found that MrlI1 and MrlI2 could synthesize distinct N-acyl homoserine lactone (AHL) autoinducers in rich medium cultures, and the expression of mrlI1 was shown to be growth-phase-dependent. MrlI3 did not produce any detectable AHL molecules under the culture conditions tested. To investigate whether these AHL synthases affect nodulation, we examined the nodulation of AHL-deficient mutants on their native plant host Lotus corniculatus and found that the efficiency of nodulation of bacteria with mutations of any of these 3 synthase genes was reduced, suggesting that quorum sensing systems in M. loti may play an important role in successful establishment of rhizobium-legume symbiosis. PMID- 19295656 TI - Isolation and identification of tyramine-producing enterococci from human fecal samples. AB - Lactic acid bacteria (LAB) are recognized as a group of important microorganisms because of their crucial role in food fermentation and their contribution to the maintenance of health homeostasis, as natural inhabitants of the human mucosa. However, the metabolic activities of some strains, such as the ability to synthesize biogenic amines (BAs), can be detrimental to human health. BAs are low molecular weight compounds synthesized by the enzymatic decarboxylation of amino acids. Tyramine, one of the most biologically active BAs, is produced by certain strains of LAB related to food fermentations. Since no data are available as to whether tyramine originates exclusively from food intake, or, like polyamines, could be formed by gut bacteria, this study was focused on the isolation of tyramine-producing LAB from human feces. Different strains of Enterococcus faecium and Enterococcus faecalis able to produce tyramine in culture conditions were isolated. PMID- 19295657 TI - Novel LPL mutations associated with lipoprotein lipase deficiency: two case reports and a literature review. AB - Lipoprotein lipase (LPL) is a key enzyme involved with hydrolysis and removal of triglycerides from plasma. LPL deficiency is a rare condition with an estimated prevalence of 1 in 106. It is characterized biochemically by elevated triglycerides and lowered HDL in the plasma and clinically by a constellation of signs and symptoms during childhood including failure to thrive, lipemia retinalis, eruptive xanthomas, hepatosplenomegaly, and acute pancreatitis. Nearly 100 mutations in the LPL gene have been associated with LPL deficiency. Here we report 2 unrelated pedigrees with LPL deficiency from 2 novel disease-causing LPL mutations: a Gly159Glu missense mutation in exon 5 and a 4-bp ACGG deletion at the 3' boundary of exon 2. We present molecular findings of these 2 cases and review the biochemical, clinical, and genetic features of LPL deficiency. PMID- 19295658 TI - Impact of maternal undernutrition on diabetes and cardiovascular disease risk in adult offspring. AB - Epidemiological, clinical, and experimental observations have led to the hypothesis that the risk of developing chronic diseases in adulthood is influenced not only by genetic and adult lifestyle factors, but also by environmental factors during early life. Low birth weight, a marker of intrauterine stress, has been linked to predisposition to cardiovascular disease (CVD) and diabetes. The compelling animal evidence and significant human data to support this conclusion are reviewed. Specifically, the review discusses the role of maternal nutrition before and during pregnancy, placental insufficiencies and epigenetic changes in the increased predisposition to diabetes and CVD in adult life. The impact of low birth weight and catch-up growth as they pertain to risk of disease in adult life is also discussed. In addition, adult disease risk in the overnourished fetus is also mentioned. Reference is made to some of the mechanisms of the induction of diabetes and CVD phenotype. It is proposed that fetal nutrition, growth and development through efficient maternal nutrition before and during pregnancy could constitute the basis for nutritional strategies for the primary prevention of diabetes and CVD. PMID- 19295659 TI - Extraction of membrane cholesterol disrupts caveolae and impairs serotonergic (5 HT2A) and histaminergic (H1) responses in bovine airway smooth muscle: role of Rho-kinase. AB - Some receptors and signaling molecules, such as Rho-kinase (ROCK), localize in caveolae. We asked whether the function of histamine receptors (H(1)) and 5 hydroxytryptamine (serotonin) receptors (5-HT(2A)) in bovine tracheal smooth muscle are modified after caveolae disruption and if so, whether the altered ROCK activity plays a role in this modification. Methyl-beta-cyclodextrin (MbetaCD), used to deplete membrane cholesterol, was shown to disrupt caveolae and diminish sustained contractions to histamine (approximately 80%), 5-HT (100%), alpha methyl-5-HT (100%), and KCl (approximately 30%). Cholesterol-loaded MbetaCD (CL MbetaCD) restored the responses to KCl and partially restored the responses to agonists. ROCK inhibition by Y-27632 diminished contractions to histamine (approximately 85%) and 5-HT (approximately 59%). 5-HT or histamine stimulation augmented ROCK activity. These increases were reduced by MbetaCD and partially reestablished by CL-MbetaCD. The increase in intracellular Ca(2+) that was induced by both agonists was reduced by MbetaCD. The presence of caveolin-1 (Cav 1), H1, 5-HT(2A), and ROCK1 was corroborated by immunoblotting of membrane fractions from sucrose gradients and by confocal microscopy. H(1) receptors coimmunoprecipitated with Cav-1 in caveolar and noncaveolar membrane fractions, whereas 5-HT(2A) receptors appeared to be restricted to noncaveolar membrane fractions. We conclude that caveolar and cholesterol integrity are indispensable for the proper functionality of the H(1) and 5-HT(2A) receptors through their Rho/ROCK signaling. PMID- 19295661 TI - Effect of stimulation frequency on force, net power output, and fatigue in mouse soleus muscle in vitro. AB - The effects of electrical stimulation frequency on force, work loop power output, and fatigue of mouse soleus muscle were investigated in vitro at 35 degrees C. Increasing stimulation frequency did not significantly affect maximal isometric tetanic stress (overall mean +/- SD, 205 +/- 16.6 kN.m-2 between 70 and 160 Hz) but did significantly increase the rate of force generation. The maximal net power output during work loops significantly increased with stimulation frequency: 18.2 +/- 3.7, 22.5 +/- 3.3, 26.8 +/- 3.7, and 28.6 +/- 3.4 W.kg(-1) at 70, 100, 130, and 160 Hz, respectively. The stimulation frequency that was used affected the pattern of fatigue observed during work loop studies. At stimulation frequencies of 100 and 130 Hz, there were periods of mean net negative work during the fatigue tests due to a slowing of relaxation rate. In contrast, mean net work remained positive throughout the fatigue test when stimulation frequencies of 70 and 160 Hz were used. The highest cumulative work during the fatigue test was performed at 70 and 160 Hz, followed by 130 Hz, then 100 Hz. Therefore, stimulation frequency affects power output and the pattern of fatigue in mouse soleus muscle. PMID- 19295660 TI - Reduced in vivo phosphodiesterase-4 response to acute noradrenaline challenge in diet-induced obese rats. AB - Altered sympathetic nervous activity has been linked to the development and persistence of obesity, partly relating to overfeeding. Binding of the selective, positron-emitting phosphodiesterase-4 (PDE4) inhibitor (R)-[11C]rolipram provides a direct index of the cAMP-hydrolyzing enzyme PDE4. This study examines progressive alterations in PDE4 in a high-fat-fed obese animal model. (R) [11C]Rolipram was injected into diet-induced obese (DIO) and diet-resistant (DR) rats; the animals were killed after 45 min, tissues were extracted, and radioactivity was quantified. Responsiveness of PDE4 to acute noradrenaline (NA) stimulation was determined by 3 h pretreatment with the NA reuptake inhibitor desipramine. There was minimal variance in caloric intake, weight gain, fasting glucose, insulin, and energy expenditure (indirect calorimetry) measures. Basal (R)-[11C]rolipram binding was comparable between DIO and DR rats at 2 or 8 weeks of feeding. The normal increase of PDE4 levels in response to elevated NA by desipramine pretreatment was ablated in PDE4-rich tissues, including brain, heart, and skeletal muscle, of DIO animals after 8 weeks of high-fat diet. Lean DR rats maintained PDE4 responsiveness indicative of a normal NA signal transduction. PMID- 19295662 TI - Comparison of anorexia, lethargy, and fever induced by bacterial and viral mimetics in rats. AB - Although it has been established that some acute phase responses present differently depending on whether a virus or bacteria activates the innate immune system, it has not yet been established whether fever and sickness behaviors, such as anorexia and lethargy, present differently. We therefore investigated the effects of administering lipopolysaccharide (LPS) and polyinosinic : polycytidylic acid (poly I:C) on body temperature, food intake, body mass, and activity (cage activity and wheel running). Male Sprague-Dawley rats were randomly assigned to receive an intraperitoneal injection of one of LPS (75 microg/kg or 250 microg/kg), poly I:C (3000 microg/kg or 4000 microg/kg), or saline. Administration of LPS or poly I:C induced fever, anorexia, and lethargy. Although voluntary wheel running and cage activity were both significantly reduced after administration of LPS or poly I:C, they were not affected equally. Indeed voluntary wheel running was decreased on average by approximately 30% more than cage activity regardless of the dose or type of mimetic administered. Our results indicate that poly I:C is less effective at inducing anorexia, lethargy, and fever in rats than is LPS, and that avoidance of exercise in animals and humans during infection is likely to be a more prominent feature of illness than is avoidance of routine daily activity. PMID- 19295663 TI - Role of protein kinase C beta2 activation in TNF-alpha-induced human vascular endothelial cell apoptosis. AB - The circulatory inflammatory cytokine tumor necrosis factor alpha (TNF-alpha) is increased in pathologic conditions that initiate or exacerbate vascular endothelial injury, such as diabetes. Protein kinase C (PKC) has been shown to play a critical role in TNF-alpha-induced human endothelial cell apoptosis. However, the relative roles played by specific isoforms of PKC in TNF-alpha induced human endothelial cell apoptosis have not been addressed. We investigated the effects of a selective PKCbeta(2) inhibitor (CGP53353) on TNF-alpha-induced apoptosis in human vascular endothelial cells (cell line ECV304) and on the production of reactive oxygen species and nitric oxide, and compared its effects with rottlerin, a reagent that has been shown to reduce PKCdelta protein levels. Cultured human vascular endothelial cells (ECV304) were treated for 24 h with one of 4 regimes: 40 ng/mL TNF-alpha alone (TNF-alpha), TNF-alpha with 10 micromol/L rottlerin (T+rottlerin), TNF-alpha with 1 micromol/L CGP53353 (T+CGP), or untreated (control). Cell viability was measured by MTT assay, and cell apoptosis was assessed by flow cytometry. TNF-alpha-induced endothelial cell apoptosis was associated with dramatic increases in production of intracellular hydrogen peroxide (approximately 20 times greater than control) and superoxide (approximately 16 times greater than control), as measured by dichlorofluorescein and dihydroethidium fluorescent staining, respectively. This increase was accompanied by reduced activity of superoxide dismutase and glutathione peroxidase and, subsequently, an increase in the lipid peroxidation product malondialdehyde. CGP53353, but not rottlerin, abolished or attenuated all these changes. We conclude that PKCbeta(2) plays a major role in TNF-alpha-induced human vascular endothelial cell apoptosis. PMID- 19295664 TI - Multiple Class Segmentation Using A Unified Framework over Mean-Shift Patches. AB - Object-based segmentation is a challenging topic. Most of the previous algorithms focused on segmenting a single or a small set of objects. In this paper, the multiple class object-based segmentation is achieved using the appearance and bag of keypoints models integrated over mean-shift patches. We also propose a novel affine invariant descriptor to model the spatial relationship of keypoints and apply the Elliptical Fourier Descriptor to describe the global shapes. The algorithm is computationally efficient and has been tested for three real datasets using less training samples. Our algorithm provides better results than other studies reported in the literature. PMID- 19295665 TI - Freedom of Speech in Government Science. PMID- 19295666 TI - Plant resistance to nematodes: symposium introduction. PMID- 19295667 TI - Phytoalexins and their Role in the Resistance of Plants to Nematodes. AB - Phytoalexins are antibiotic compounds synthesized in an infected plant in response to infection. Nematodes are capable of eliciting phytoalexins in resistant plants. Resistant lima bean (Phaseolus lunatus) infected by Pratylenchus penetrans produces the phytoalexin coumestrol; soybean (Glycine max) infected by Meloidogyne incognita produces glyceollin; cotton (Gossypium hirsuturn) infected by M. incognita produces terpenoid aldehydes. PMID- 19295668 TI - Potential of tissue culture for breeding root-knot nematode resistance into vegetables. AB - Plant protoplast technology is being investigated as a means of transferring root knot nematode resistance factors from Solanum sisymbriifolium into the susceptible S. melongena. Solanum sisymbriifolium plants regenerated from callus lost resistance to Meloidogyne javanica but retained resistance to M. incognita. Tomato plants cloned from leaf discs of the root-knot nematode resistant 'Patriot' were completely susceptible to M. incognita, while sections of stems and leaves rooted in sand in the absence of growth hormones retained resistance. Changes in resistance persisted for three generations. It is postulated that the exogenous hormonal constituents of the culture medium are modifying the expression of genetic resistance. PMID- 19295669 TI - Current status of breeding for resistance to insects. PMID- 19295670 TI - Plant resistance in nematode pest management. AB - Plant resistance is a primary component in the development of nematode pest management programs. A survey of resistant cultivar and rootstock implementation in 75 annual and perennial crops grown in California provides an insight into the current status of plant resistance in modern agriculture. As new sources of commercially suitable resistant germplasm are made available, their successful implementation will require information on quantitative relationships of injury tolerance and hosting ability, and on relationships to intraspecific pathotypes or races present in the major agricultural regions of implementation. Computer simulation and modeling as an aid in developing these programs is discussed. A format for the implementation of a resistant line with good agronomic potential is outlined. PMID- 19295671 TI - Characterization of sialyl and galactosyl residues on the body wall of different plant parasitic nematodes. AB - The plant parasitic nematodes Helicotylenchus multicinctus, Meloidogyne javanica, Tylenchulus semipenetrans, and Xiphinema index, differing in their host specificity and parasitic habits, were analyzed as to their cuticle surface sialyl, galaclosyl, and/or N-acetylgalactosaminyl residues. The procedure involved the selective oxidation of sialic acid and galactose/N-acetylgal actosamine residues using periodate and galactose oxidase, respectively, to form reactive aldehyde groups. These functional groups were coupled directly with a new hydrazide-containing compound, the fluorescent reagent lissamine rhodamine beta-alanine hydrazide, or they were utilized to introduce DPN-groups to the nematode cuticle. The distribution of the DNP-tagged glycoconjugates was visualized by treating the nematodes with rabbit anti-DNP antibody and staining with fluorescein isothiocyanate (FITC)-labeled goat antirabbit IgG. Sialo residues were observed along the entire outer body wall of the first three aforementioned nematodes, but there were some differences in reaction among the various life stages within the species. In X. index, sialo residues were sited in the tail and head areas, mainly on the lips, oral opening, amphid apertures and stylet. Galactose oxidase treatments revealed galactose on N-acytylgalactosamine residues on T. sentipenetrans and X. index, but there were no indications that their presence was dependent on the developmental stage. Trypsin, pronase, and neuraminidase pretreatment completely abolished the fluorescence in T. semipenetrans but did not alter the sialo residue binding reaction in H. multicinctus or M. javanica, indicating possible differences in the outer body wall saccharide structure and composition between these nematodes. The existence and nature of sugar residues on the cuticle surface of nematodes could contribute to an understanding of the specific recognition by phytophagous nematodes of their host, and perhaps also of the virus transmission mechanism in those nematodes which serve as vectors. PMID- 19295672 TI - Localization of Cuticular Binding Sites of Concanavalin A on Caenorhabditis elegans and Meloidogyne incognita. AB - Utilizing a Concanavalin A (Con A)-hemocyanin conjugate, the majority of cuticular Con A binding sites were shown to be localized on the head region of Caenorhabclitis elegans and Meloidogyne incognita. Secretions which apparently emanated from the amphids and inner labial papillae did not label. PMID- 19295673 TI - Population Dynamics of Meloidogyne incognita on Corn Grown in Soil in Fested with Arthrobotrys conoides. AB - Microplot and greenhouse experiments were conducted to evaluate the effects of soil incorporation of the nematophagous fungus Arthrobotrys conoides and green alfalfa mulch on the population dynamics of Meloidogyne incognita on corn. Reproduction of M. incognita and the incidence of root galling were reduced by the addition of A. conoides and/or green alfalfa in all tests. Numbers of juveniles were reduced by as much as 84%, and eggs were fewest in early to mid season soil samples from microplots. Yields increased in treatments with A. conoides and/or green alfalfa in greenhouse tests and in the microplot tests in 1979. No interaction was found between the fungus and green alfalfa in the reduction of the nematode population. PMID- 19295674 TI - Relationship Between Meloidogyne hapla Density and Damage to Carrots in Organic Soils. AB - Field and growth chamber experiments were conducted to determine the effect of five initial densities (Pi - 20 - 240/100 cm(3) soil) of Meloidogyne hapla on carrot development and yield of storage roots at maturity. Carrots growing in infested and noninfested organic soil were harvested after 15, 29, 44, 59, and 106 days of growth in controlled environment chambers and after l l 0 days in field plots. Nematodes affected weight of roots and foliage, weight and length of the storage portion of tap roots, and induced malformations (forking), galling, and hairiness of tap roots. In most cases the data could not be represented satisfactorily by the exponential model of Seinhorst: y = m + (l-m) Z(P-t). In growth chambers the weight of mature storage roots was not correlated to initial nematode density, but there was a significant negative correlation between weight of storage roots and initial nematode density in field plots. Tolerance levels were calculated as points where the regression lines reached the growth level on noninoculated plants. The tolerance levels of foliage were higher than those of roots, and increased with age of plants. The tolerance level of marketable weight in field plots, average crop value, and a hypothetical control cost function are used to discuss the possibility of optimizing chemical control of root-knot nematode in organic soils. PMID- 19295675 TI - Varietal Response of Tomato to the Interaction of Salinity and Meloidogyne incognita Infection. AB - Response of tomato (Lycopersicon esculentum) cultivars to a range of conductivity levels was tested in the presence and absence of Meloidogyne incognita. The conductivity levels were produced by appropriate adjustment of a 1:1 solution of sodium chloride and calcium chloride. The growth of M. incognita resistant ('Beefmaster' and 'Atkinson') and susceptible ('Hunts 2580' and 'Ronita') tomato plants was inversely related to soil salinity between EC(e) 0 and 5 mmhos/cm. Nematode inoculation of salt-stressed plants significantly reduced plant height, fresh and dry weight, number of flowers, and fruit weight in most cultivars. In Hunts 2580, flower number and fruit weight increased; apparently flower production shifted from determinate to indeterminate, with negative implications for mechanical harvesting. Nematode reproduction on susceptible varieties also decreased with increase in salinity. PMID- 19295676 TI - Effects of Management Practices on Meloidogyne incognita and Snap Bean Yield. AB - Phenamiphos applied at 6.7 kg ai/ha through a solid set or a center pivot irrigation system with 28 mm of water effectively controlled root-knot nematodes, Meloidogyne incognita, and resulted in greater snap bean growth and yields irrespective of growing season, tillage method, or cover crop system. The percentage yield increases attributed to this method of M. incognita control over nontreated controls were 45% in the spring crop, and 90% and 409% in the fall crops following winter rye and fallow, respectively. Root galling was not affected by tillage systems or cover crop, but disk tillage resulted in over 50% reduction in bean yield compared with yields from the subsoil-bed tillage system. PMID- 19295677 TI - Nematode temperature responses: a niche dimension in populations of bacterial feeding nematodes. AB - The optimum temperatures for population development were determined for six species of bacterial-feeding nematodes from among eight temperatures, ranging from 5 to 40 C. Four of the species are cohabiting species. The range of temperatures over which population development occurs (temperature niche breadth) is different for the cohabiting species. This difference may be a means of reducing competition between species, thus increasing temperatures over which habitats can be exploited. PMID- 19295678 TI - Chloroplast Differentiation in Tomato Root Galls Induced by the Root Knot Nematode Meloidogyne incognita. AB - Primary roots of tomato, Lycopersicon esculentum cv. Marglobe, were cultured aseptically on agar containing a standard nutrient formulation with or without kinetin. When secondary roots developed, cultures were inoculated with the root knot nematode, Meloidogyne incognita. Following inoculation, the cultures were divided into two groups which were incubated either in total darkness or in 16-h light-8-h dark cycles. At 24 h, 1, 2, 3, and 4 wk after incubation, roots from all cultures were processed for transmission electron microscopy. Fine structural observation of the parenchyma tissue in galls from the inoculated cultures indicated that starch containing plastids or amyloplasts, which are usually present and remain undifferentiated in these root cells, developed into chloroplasts. These chloroplasts contained a membrane system indistinguishable from those found in leaves of intact plants. Although plastid development was not affected when uninoculated cultures were incubated in the light, differentiation of the amyloplast was induced when roots were cultured on the medium containing kinetin. These results suggest that plastid differentiation in the inoculated tissue may be influenced by an accumulation of kinetin in the gall, which is induced by the nematode and serves as the nutrient sink for its feeding. PMID- 19295679 TI - Meloidogyne platani n. sp. (Meloidogynidae), a Root-knot Nematode Parasitizing American Sycamore. AB - Meloidogyne platani n. sp. is described and illustrated from specimens obtained from roots of American sycamore, Platanus occidentalis, in Virginia. This new species shows certain similarities with M. arenaria but differs from it by a number of distinctive characters. The perineal pattern of females is rounded with fine, wavy to zig-zag striae and raised, convoluted striae in the inner lateral line regions. The stylet of females is 16.5 mum long with large, rounded stylet knobs set off from the shaft. Males have a low head cap and smooth head region. The styler length is 22.0 mum, and the stylet knobs are rounded and set off from the shaft. Mean second-stage juvenile length is 443.0 mum, and stylet length is 12.2 mum. The head region of juveniles is not annulated, and the tail has a definite terminus. This nematode causes severe galling and reproduces well on sycamore. Other good hosts include white ash and tobacco cv. NC 95. M. platani n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of approximately 45 (2n). PMID- 19295680 TI - Macroposthonia sicula n. sp. (Nematoda: Criconematidae), a Parasite of Olive Trees in Sicily. AB - Macroposthonia sicula n.sp. collected from rhizosphere and roots of olive (Olea europaea L.) at Kamarina, Sicily, Italy, is described and illustrated. It is distinguished from the related species (M. sphaerocephala and M. maskaka) by the longer styler and the characteristic narrowing postvulval portion of the body. PMID- 19295681 TI - Effect of Age of Alfalfa Root on Penetration by Pratylenchus penetrans. AB - Penetration by all migratory life stages of Pratylenchus penetrans into roots of alfalfa (Medicago sativa L. cv. Du Puits) was inversely proportional to tissue age. Two-day-old tissue in the root hair zone was penetrated twice as much as 10- or 20-day-old sections of the tap root. Age-related differences were also observed in branch roots; these differences were not affected by increasing the number of nematodes from 1 to 10 per inoculation site, nor by increasing the length of the incubation period from 6 to 96 h. Age-related differences were only significant with 3-wk-old plants, not with 2- and 1-wk-old seedlings. Nematodes entered roots at temperatures from 5 to 30 C with maximum entry at 20 C and minimum at 5 C. At all temperatures, except 5 C, penetration into young tissue (2 days) was significantly greater than into medium (10 days) and old (20 days) tissue. Females and third-stage larvae entered the different-aged root sections 122% and 83%, respectively, more than did males. Two-day-old seedlings of the alfalfa cultivars Vernal, Saranac, and Du Puits were penetrated equally by P. penetrans. Perhaps the inverse relationship between penetration and age of root is, in part, responsible for the increasing resistance or tolerance of plants to nematode damage as they grow older. PMID- 19295682 TI - Interactions among selected endoparasitic nematodes and three pseudomonads on alfalfa. AB - Meloidogyne hapla, Pratylenchus penetrans, and Helicotylenchus dihystera, reduced the growth of 'Saranac AR alfalfa seedlings when applied at concentrations of 50 nematodes per plant. All except P. penetrans reduced seedling growth when applied at 25 per seedling. M. hapla reduced growth when applied at 12 per seedling. Nematodes interacted with three pseudomonads to produce greater growth reductions than were obtained with single pathogens, suggesting synergistic relationships. Ditylenchus dipsaci, applied at 25 or 50 nematodes per seedling, reduced plant weight compared with weights of control plants, but did not interact with test bacteria. All of the nematodes except D. dipsaci produced root wounds which were invaded by bacteria. PMID- 19295683 TI - Partitioning yield loss on yellow squash into nematode and insect components. AB - The effect of a contplex of several insect and nematode pests on yield of yellow squash (Cucurbita pepo L.) was examined in two field tests in southern Florida. Applications of permethrin for insect control and oxamyl primarily for nematode control plus some insect control were made alone and in combination to achieve differential reduction of various insect and nematode components contributing to yield loss. The effect of these components on yield was further analyzed by multiple regression. Yield losses in weight of small fruit to nematode and insect pests together were estimated at 23.4% and 30.4% in each of the two tests, respectively. In the first test, this loss was attributed to the melonworm, Diaphania hyalinata, while in the second test, it was attributed to D. hyalinata and the nematodes Quinisulcius acutus and particularly Rotylenchulus reniforrnis. D. hyalinata accounted for further losses of 9.0% and 10.3%, respectively, from direct damage to the fruit. Despite the presence of low levels of Diabrotica balteata, Liriomyza sativae, and Myzus persicae, yields were little affected by these pests. Prediction of yield loss by multiple regression analysis was more accurate when both insect and nematode populations were present in the plots than when nematodes alone were present. PMID- 19295684 TI - Nematicidal principles from two species of lamiaceae. AB - Aqueous extracts of Ocimum sanctum and O. basilicum leaves contained compounds that killed Meloidogyne incognita larvae in 160 min. Thin layer and gas-liquid chromatography, and infrared spectrophotometry indicated that the essential oils eugenol and linalool were the active nematicidal compounds. PMID- 19295685 TI - An Improved Method for Long-Distance Shipping the Mosquito Parasite Romanomermis culicivorax. AB - To prevent losses during long-distance shipment, a new delivery system was developed for the mosquito parasite Romanomermis culicivorax. A specially designed shipping container, two types of culture media (sand and a mixture of sand, vermiculite, and charcoal [SVC]), and two types of insert (polystyrene and polyester urethane foam [PUF]) were tested. Cultures shipped in SVC/PUF insert combination produced significantly higher yields of preparasites than did the other three combinations tested, and the sand/polystyrene combination produced significantly lower yields than did either sand or SVC media shipped with the PUF insert. Yields from cultures shipped in sand/PUF and SVC/PUF combinations were significantly lower than from unshipped controls. Maximum losses (52%) occurred in cultures shipped when 13-16-wk old. Also, yields of preparasites were significantly lower from cultures shipped singly when compared to similar cultures shipped in groups of eight. Techniques and procedures developed in this study did not completely solve the loss associated with delivery of R. culicivorax. For the first time, however, the system can guarantee the delivery of a quality product. The study defines the importance of the carrier medium, age of cultures, shipment size, and the restriction of the movement of the culture medium. PMID- 19295686 TI - In vitro Cultivation of the Entomogenous Nematode Filipjevimermis leipsandra. AB - The mermithid nematode, Filipjevimermis leipsandra, was successfully cultivated to the preadult stage in Schneider's Drosophila medium supplemented with 20% fetal bovine serum. Upon transfer to a solid substrate the preadults continued to develop into ovipositing adult females. Four molts were observed. The first molt occurred in the egg. The second occurred after 6-8 days in culture during which the very thin cuticle was shed completely. The third molt occurred after 18-20 days in culture; the cuticle was retained by the third-stage nematode. This stage was considered comparable to the preadult stage that emerges from host larva, Diabrotica spp. The fourth molt occurred within 12 days after the preadult was transferred from the liquid medium to a solid substrate. Adult females began ovipositing viable eggs 1-3 days after the final molt. PMID- 19295687 TI - Type specimens on deposit at university of california davis nematode collection. AB - A list of the deposited type specimens, historical highlights, and maintenance procedures are given for the type section of the University of California Davis Nematode Collection (UCDNC). The type specimen section contains more than 630 species and 10,740 individual specimens mounted on more than 3,600 microscope slides. This list of deposited types is only a type specimen location reference and should not be used for type status or as a check list. PMID- 19295688 TI - Development of Meloidogyne incognita in Stored Potato Tubers. PMID- 19295689 TI - Anhydrobiosis in Nematodes II: Carbohydrate and Lipid Analysis in Undesiccated and Desiccate Nematodes. AB - Glycogen, trehalose, glucose, and total lipid contents of six nematode species were studied. Anhydrobiotic Anguina tritici and Ditylencbus dipsaci stored trehalose in preference to glycogen and only small amounts of glucose were detected. Glycogen content was also reduced in anhydrobiotic Aphelenchus avenae. Conversely, Panagrellus redivivus and Turbatrix aceti contained large amounts of glycogen, appreciable amounts of glucose, and minimal amounts of trehalose. Ditylenchus myceliophagous "curds" contained low amounts of glycogen and very little trehalose; total lipid was 60% of that in fresh samples. The lipid contents of fresh samples of P. redivivus, T. aceti, and A. avenae were high (23.1, 21.9, and 36.7% dry weight, respectively), but in anhydrobiotic A. avenae larvae the level was reduced by over 60%. In contrast, lipid levels remained high in anhydrobiotic A. tritici and D. dipsaci larvae (40.6 and 38.3%, respectively). Analysis of lipid composition in anhydrobiotic A. tritici and A. avenae did not indicate any specific metabolic adaptations to desiccation survival. PMID- 19295690 TI - Biology of Meloidogyne platani Hirschmann Parasitic on Sycamore, Platanus occidentalis. AB - The development of Meloidogyne platani on sycamore was followed for 40 days (22 28 C). Juveniles penetrated the feeder roots behind the root cap and invaded the vascular cylinder within 3 days after inoculation. All subsequent development of the nematodes and host effects occurred only within the stele. The second juvenile molt and sex differentiation occurred by the 17th day. Young females were observed by the 26th day. Eggs were observed inside the roots by the 35th day and were exposed to the surface of galls by the 40th day. In pathogenicity studies, a significant negative correlation was shown to exist between fresh shoot and root weights and inoculum density. Besides sycamore, white ash was the only hardwood species tested to become infected. Of the herbacious plants tested, tobacco was heavily galled, tomato and watermelon moderately galled, and pepper only slightly galled. Egg production was moderate on tobacco, slight on tomato and watermelon, and absent on pepper. PMID- 19295691 TI - The Relationship Between Soil Populations of Meloidogyne incognita and Yield Reduction of Soybean in the Coastal Plain. AB - In a replicated field plot experiment, the population density of Meloidogyne incognita was monitored biweekly through the overwintering period (December through April) between soybean crops. The population survived as second-stage juveniles whose numbers remained stable through the winter months and did not decline until February. The yields of plots planted with a M. incognita susceptible cultivar were negatively correlated with the numbers of juveniles recovered at all preplanting sampling dates. In the mid-winter period (December through February), a regression equation describing the relationship predicted a yield reduction (slope) equivalent to 5.36 kg/ha for each juvenile in a 10-cm(3) soil sample. In two subsequent field experiments, conducted in different sites and years, mid-winter (November) sampling gave yield reduction predictions of 4.65 and 6.69 kg/ha. Tests of the null hypothesis gave no evidence to indicate that the three slopes differed (P = 0.05). A regression analysis of combined data from the three experiments determined a mid-winter predictive yield reduction of 5.31 kg/ha for each juvenile in the 10-cm(3) sample. As the sampling time approached the planting date, there were changes in the predictive yield reductions due to each juvenile in a sample. These are best described by the equation, gamma (yield loss) = 54.47 - 0.67X + 0.0023X(2), where X equals the days remaining between sampling and planting. Soil sampling should be performed during mid-winter (November through January) for the most reliable prediction of soybean yield loss. PMID- 19295692 TI - The Effect of Arthrobotrys conoides on Meloidogyne incognita Population Densities in Corn as Influenced by Temperature, Fungus Inoculum Density, and Time of Fungus Introduction in the Soil. AB - In greenhouse experiments, the effect of Arthrobotrys conoides on Meloidogyne incognita population densities as affected by soil temperature, inoculum density, and green alfalfa was determined. The effect on M. incognita population densities was greater at a soil temperature of 25 C than at 18 or 32 C. Nematode control by A. conoides was most effective when the fungus was introduced into the soil 2 wk prior to nematode inoculation and planting of corn. Inoculum density of A. conoides was positively correlated with plant shoot weight (r = 0.81) and negatively correlated with numbers of Meloidogyne juveniles (r = -0.96), eggs ( 0.89) and galls per gram of root (-0.91). A. conoides was not isolated from green alfalfa, but was isolated from alfalfa-amended soil to which no fungus had been added. PMID- 19295693 TI - Differences in the Response of Certain Weed Host Populations to Heterodera schachtii. AB - Significant differences (P = 0.05) in nematode reproduction were observed among populations of Heterodera schachtii and weed collections of black nightshade, common lambsquarters, common purslane, redroot-pigweed, shepherdspurse, and wild mustard from Colorado, Idaho, Oregon, and Utah. Colorado weeds supported the greatest nematode development (P = 0.05). Weeds collected from Idaho and Utah were similar with respect to their response to H. schachtii with the exception of shepherdspurse. At increasing soil temperatures, a Utah redroot-pigweed collection showed a higher percent susceptibility to a Utah nematode population than to nematode populations from the other states (P = 0.05). There was a higher percentage of susceptible plants when the weed host population was collected from the same geographical area as the nematode inoculun. PMID- 19295694 TI - Pathological Interaction of a Combination of Heterodera schachtii and Meloidogyne hapla on Tomato. AB - Increased culturing of a tomato population of Heterodera schachtii (UT1C) on tomato for 480 days (eight inoculation periods of 60 days each) significantly increased virulence to 'Stone Improved' tomato. A synergistic relationship existed between Meloidogyne hapla and H. schaehtii on tomato. A combination of H. schachtii (UTIC) and M. hapla significantly reduced tomato root weights by 65, 64, and 61% below root weights of untreated controls, and single inoculations of M. hapla and H. schachtii, respectively. This corresponded to root reductions of 42, 44, and 46% from a combination of H. schachtii (UT1B) and M. hapla. Antagonism existed between H. schachtii and M. hapla with regard to infection courts and feeding sites. The root-knot galling index dropped from 6.0 with a single inoculation of M. hapla to 4.3 and 3.3 with combined inoculations of M. hapla plus UT1B and M. hapla plus UTIC cyst nematode populations. The pathological virulence of H. schachtii to sugarbeet was not lost by extended culturing on tomato; there were no differences in penetration, maturation, and reproduction between sugarbeet populations continually cultured on sugarbeet and the population continually cultured on tomato. PMID- 19295695 TI - Morphometric and serologic comparisons of a number of populations of cyst nematodes. AB - Thirty-five populations of Heterodera glycines and populations of 15 other Heterodera, Globodera, and Punctodera species were studied morphometrically and some were compared serologically. There was a wide range of each measurement within each nematode population. Except for one soybean cyst nematode population from Indiana, which was a tetraploid and considerably larger than the others, morphometric measurements overlapped. In a discriminant function comparison most of the populations were closely grouped but at least three were rather distinctly separated. Morphometrically H. fici, H. cruciferae, H. schachtii, and H. trifolii were closely associated with H. glycines. Serology indicated a close relationship between H. glycines, H. lespedezae, H. trifolii, H. schachtii, and the Heterodera sp. from Rumex, while H. betulae appeared to be more distantly related. PMID- 19295696 TI - Tolerance Limit of the Sugarbeet to Heterodera schachtii. AB - Field and greenhouse experiments showed that yield losses of sugarbeet, Beta vulgaris, did not occur in soil infested with fewer than eight Heterodera schachtii eggs/g soil. However, larger population densities greatly reduced sugarbeet yield. In the field experiment, the yield in microplots inoculated with more than 64 eggs/g soil was less than 20% of yields in uninoculated microplots. Nevertheless, tolerance limits of 4 and 1.8 eggs/g soil, in greenhouse and field microplots, respectively, were derived by fitting the data with the equation y =m + (l - m)z(P-T). Maximum rates of multiplication of 55 and more than 300, and equilibrium densities of 340 and 130 eggs/g soil, were estimated in greenhouse and field microplot tests, respectively. PMID- 19295697 TI - Effects of Host Variety, Photoperiod and Chemical Treatments on Hatching of Globodera rostochiensis. AB - Golden nematode cysts were collected after they had completed one generation on an early, a mid-season, or a late-maturing potato variety ('Superior,' 'Katahdin,' and 'Sebago,' respectively). The plants were grown in 8.5- or 16-h photoperiods and treated with gibberellic acid (GA) or xylocaine or both. When treated subsequently with potato root-diffusate few (13.3%) juveniles hatched from cysts collected from Sehago, more (22.3%) from cysts collected from Katahdin, and most (36.0%) from cysts collected from Superior. Hatching was greater from cysts collected from plants treated with GA than from untreated plants. Short photoperiods decreased hatching from cysts collected from Katahdin but increased hatching from cysts collected from Superior and Sebago. Gibberellic acid had no detectable effect on hatching, but EDTA had some. PMID- 19295698 TI - Effect of Foliar Applications of Carbofuran and a Related Compound on Plant parasitic Nematodes under Microplot and Field Conditions. AB - Studies were conducted to investigate the basipetal translocation of nematicidal activity from foliar treatments of carbofuran and its analog, 2,3-dihydro-2,2 dimethyl-7-benzofuranyl ([dibutylamino]thio)methyl carbamate, on corn in microplot studies and on tobacco and potato in field trials. Two and three foliar applications of either product at 2,400 mug/ml (9-20 Kg ai/Ha) significantly reduced populations of Pratylenchus penetrans in roots and populations of Tylenchorhynchus claytoni, Xiphinema americanum, and Hoplolaimus sp. in soil. In most cases there was no difference in control between two or three chemical applications. Foliar treatments with carbofuran were equivalent to, or better than, soil treatment, although rates of applications were different. PMID- 19295699 TI - Granular Nematicides for Control of the Yam Nematode, Scutellonema bradys, and Relevant Residues in Raw Tubers. AB - Four granular nentaticides were evaluated for control of the yam nematode, Scutellonema bradys (Steiner &LeHew) Andrassy, on Guinea yam, Dioscorea rotundata Poir, under field conditions prevelant in the tropics. A single application of nematicides (sidedressing) at the rate of 2 kg ai/ha as postplanting treatment at the onset of the rainy season depressed numbers of S. bradys attacking yams during the growing season and significantly increased tuber yields over untreated. The efficacy, based on the regression coefficient values of evaluated nematicides, was in the order of miral, carbofuran, aldicarb, and oxamyl (b = 75.9, -75.5, -72.1, and -65.9, respectively). Yam tuber yields increased by 136.9, 90.6, 87.9, and 85.3% over untreated (P = 0.05) in aldicarb, carbofuran, oxamyl, and miral treated plots, respectively. Residues in raw tubers pretreated with aldicarb, carbofuran, or miral were negligible (front less than 0.02 to 0.3 ppm) and far below the established tolerance levels (l.0 and 1.3 ppm for aldicarb and carbofuran, respectively) of a related crop in the United States. This is the first report on residues of systemic pesticides in yams. PMID- 19295700 TI - Susceptibility of Soybean Cultivars and Lines to Pratylenchus hexincisus. AB - Population increase of Pratylenchus hexincisus on 41 soybean cultivars (maturity groups I-VI) and lines was tested under greenhouse conditions. After 3 months, P. hexincisus was recovered from the roots of all plants tesled. Final populations of P. hexincisus per pot were larger than the initial population in 13 cultivars. Pathogenicity of P. hexincisus on five soybean cultivars representing maturity groups (I-V) was demonstrated under greenhouse conditions. An inoculmn of 5,000 P. hexineisus/plant significantly decreased the root and shoot biomass of all five soybean cultivars after 3 months. PMID- 19295701 TI - Description of Quinisulcius solani n.sp. (Nematoda: Tylenchorhynchidae) with a Key to the Species and Data on Scutylenchus koreanus from Pakistan. AB - A new species. Quinisulcius solani, is described and illustrated from specimens on Solanum tuberosum from Murree Hills, Pakistan. Q. solani n.sp. differs from its closest relative, Q. acutus (Allen, 1955) Siddiqi, 1971, by its spiral to open 'C' shaped body and stylet length of 19 mun, vs. 17 mum in Q. acutus. In Q. acutus the stylet knobs project anteriorly but slope posteriorly in Q. solani n.sp. Tail annules number 17 in Q. acutus but 38 in Q. solani n.sp.; also the phasmids in the former species are at mid-tail and in the latter are in anterior half of tail, at about 34%. Q. solani n.sp. is also closely related to Q. capitatus (Allen, 1955) Siddiqi, 1971 but differs in some characters. Head annules number eight and stylet length is 17 mum in Q. capitatus, but head annules are six and stylet length is 19 mum in Q. solani n.sp. In Q. capitatus T/ABW is 3 vs. 2.2 in Q. solani n.sp. Also, phasmids are located at mid-tail on Q. capitatus but at ahout one-third of the tail on Q. solani n.sp. A key to the 10 species of Quinisulcius is also presented. Scutylenchus koreanus (Choi &Geraert, 1971) Siddiqi, 1979 is recorded for the first time in Pakistan and morphometric data and illustrations given. PMID- 19295702 TI - Respiratory Physiology of a Facultative Anaerobe, Romanomermis culicivorax. AB - The respiratory physiology of postparasitic larval and adult Romanornermis culicivorax was studied by manometric and polarographic methods. Endogenous respiration rates were relatively low and unaffected by postemergent development. The respiratory quotient (RQ) of larvae and young adults was 0.4 but increased to 0.7 about 3 wk after emergence. Exogenous glucose (0.02 mM) had no effect on QO or RQ. Respiration of adults and larvae was completely inhibited by KCN (l mM) but not by NaN (l mM) or 2,4-DNP (0.1 mM). The nematodes survived exposure to cyanide (l mM) for 18 h. Carbon dioxide (10%) inhibited respiration. The postparasitic stages of the nematode were mixed respiratory regulators conformers. Exposure to anaerohic conditions resulted in an increased postanaerobic oxygen consumption which persisted for 3.5 h. The experiments confirmed that the postemergent stages of the nematode are facultative anaerobes. PMID- 19295703 TI - Teratological Development in the Cephalic Anatomy of the Nematode Romanomermis culicivorax. AB - Adults of Romanomermis culicivorax obtained from mass cultures were examined by scanning and transmission electron microscopy to determine the organization of their anterior sense organs. The normal pattern apparently consists of two lateral amphids plus six cephalic papillae. Lateral cephalic papillae contain two sense organs, each with a cuticular pore, while subdorsal and subventral papillae have three sense organs, each with a cuticular pore. About 30% of females and 80% of males examined showed aberrant developments in these sense organs. Both cuticle and underlying cells (hypodermis and neurons) are affected; some sense organs are absent while others are incompletely formed. Few aberrant worms were found in a smaller collection of worms reared in lower population densities. Perhaps aberrant forms are examples of teratological development resulting from, or promoted by, conditions used for mass rearing of biological control agents. PMID- 19295704 TI - Developmental Biology of Caenorhabditis elegans: Symposium Introduction. PMID- 19295706 TI - The Cuticle of Caenorhabditis elegans. PMID- 19295705 TI - Nematode postembryonic cell lineages. AB - The complete postembryonic ceil lineages of the free-living nentatodes Caenorhabditis elegans and Panagrellus redivivus are known. Postembryonic cell divisions lead to substantial increases in the number of cells and, in most cases, in the number of types of cells in the neuronal, muscular, hypodermal, and digestive systems. The patterns of postembyronic cell divisions are essentially invariant and generate a fixed number of progeny cells of strictly specified fates. Cell fates depend upon both lineage history and cell-cell interactions: lineage limits the developmental potential of each cell and, for certain cells, cell-cell interactions specify which of a small number of alternative potential fates is acquired. Relatively simple differences in cell lineage account for some of the striking differences in gross morphology both between sexes and between species. Genetic studies indicate that these cell lineage differences reflect one or a few relatively simple mutational events. Interspecific differences in cell lineage are likely to be good indicators of evolutionary distance and may be helpful in defining taxonomic relationships. Both the techniques utilized in, and the information acquired from, studies of cell lineages in C. elegans and P. redivivus may prove useful to other hematologists. PMID- 19295707 TI - The Development and Motility of Caenorhabditis elegans Spermatozoa. PMID- 19295708 TI - Developmental Determinants in Embryos of Caenorhabditis elegans. AB - C. elegans is proving useful for the study of cell determination in early embryos. Breeding experiments with embryonic lethal mutants show that abnormal embryogenesis often results from defective gene function in the maternal parent, suggesting that much of the information for normal embryonic development is laid down during oogenesis. Analysis of a gut-specific differentiation marker in cleavage-arrested embryos has provided evidence that the potential for this differentiation behaves as a cell-autonomous internally segregating developmental determinant, which is present from the 2-cell stage onward and is partitioned into the gut precursor cell during early cleavage divisions. Visible prelocalized cytoplasmic granules that segregate with a particular cell lineage have heen observed in the embryonic germline precursor cells by fluorescent antibody staining. Whether these granules play a role in germline determina... [remainder of abstract missing in original]. PMID- 19295709 TI - Cross-mating of Romanomermis culicivorax and R. communensis (Nematoda: Mermithidae). PMID- 19295710 TI - Erratum. AB - [This corrects the article on p. 171 in vol. 13.]. PMID- 19295711 TI - Fine Structure of the Esophagus of Males of Sarisodera hydrophila (Heteroderoidea). AB - The fine structure of the esophagus, including procorpus, metacorpus, isthmus, gland lobe, and esophago-intestinal junction, is examined in males of Sarisodera hydrophila. A cuticle-lined lumen extends most of the length of the esophagus, broadens to form a pump chamber in the metacorpus, and posteriorly is continuous with junctional complexes among four esophago-intestinal cells. These four cells are partially enveloped by the gland lobe which basically consists of three gland cells, one dorsal and two subventral. Each gland cell has an anterior process which opens into the lumen of the esophagus through a cuticle-lined duct. The dorsal gland joins the lumen in the anterior portion of the procorpus, whereas ducts of the subventral glands terminate at the base of the metacorpus pump chamber. The subventral glands are predominant in the posterior portion of the gland lobe and are partially ensheathed by a narrow portion of the dorsal gland which extends to within 5 mum of the posterior terminus of the gland lobe. Contents of the dorsal gland include primarily electron dense granules, although rough endoplasmic reticulum (RER) is predominant posteriorly. Secretory granules within the subventral glands vary in morphology and are evenly distributed throughout the two ceils among other organelles, including RER and a large Golgi apparatus. Innervation of the esophagus includes nerve processes which originate from several perikaryons (cell bodies) located in the anterior portion of the gland lobe. The esophagus of males of S. hydrophila is compared with that of other Heteroderoidea, Heterodera glycines and Meloidogyne incognita. PMID- 19295712 TI - Meloidogyne cruciani n. sp. a Root-knot Nematode from St. Croix (U.S. Virgin Islands) with Observations on Morphology of This and Two Other Species of the Genus. AB - Meloidogyne cruciani n. sp. infecting tomato (Lycopersicon esculentum Mill.) in the U.S. Virgin Islands is described and illustrated. M. cruciani is distinguished from other species of the genus by having punctations around the anus of the female and by the second-stage juveniles possessing tri-lobed esophageal glands which are longer than most other species, with their posterier end at about 46.4% of the body length. The esophageal glands of the immature and adult females are contained in five separate lobes. PMID- 19295713 TI - Description of the Blueberry Root-knot Nematode, Meloidogyne carolinensis n. sp. AB - Meloidogyne carolinensis n. sp. is described from cultivated highbush blueberry (cultivars derived from hybrids of Vaccinium corymbosum L. and V. lamarckii Camp) in North Carolina. The perineal pattern of the female has a large cuticular ridge that surrounds the perivulval area, and the excretory pore is near the level of the base of the stylet. The stylet is 15.9 mum long and the knobs gradually merge with the shaft. The head shape and stylet morphology of the male are quite variable. The typical head and four variants, as well as the typical stylet and two variants, are described. The labial disc, medial lips, and lateral lips of second-stage juveniles are fused and in the same contour. The head region is not annulated. Mean juvenile length is 463.7 mum, stylet length is 11.9 mum, and tail length is 42.5 mum. PMID- 19295714 TI - Three new species of the super family neotylenchoidea (nematoda: tylenchida) from pakistan. AB - Three new species in the super family Neotylenchoidea collected in Pakistan during 1979-80 are described. New species Nothotylenchus gohleni is related to N. affinis Thorne, 1941 but is differentiated by its longer, more robust body with tine cuticular annules, a more posterior vulva, inconspicuous basal knobs of the stylet, and poorly developed metacorpal area. N. geraerti and N. tuberosus are also close to this undescribed species, hut they have a more developed metacorpal area and a larger post uterine sac. Boleodorus zaini n.sp. is distinguished by its excretory pore open at a level with the base of the posterior esophageal bulb and by six distinct lines in the lateral field. Paurodontella sohailai n.sp. is closely related to P. densa (Thorne, 1941) Hussain &Khan, 1967 and P. minuta Hussain &Khan, 1967 but differs from them by its larger body, shape of tail, and seven incisures in the lateral field. PMID- 19295715 TI - Criconematina (nematoda: tylenchida) from the aleutian islands. AB - A new genus (Cerchnotocriconenta) and three new species (C. psephinum, Hemicycliophora amchitkaensis, and Paratylenchns amundseni) are described from Adak and Amchitka Islands in the Aleutian chain. The new genus differs from all other criconematid genera in having irregular, convex sculpturing consisting of small, oval plates on the anterior and posterior regions of each annule, with the mid-annular region minutely punctate or dentate. H. arnchitkaensis n. sp. resembles H. similis Thorne and H. zuckermani Brzeski, but has only one head annule, instead of two. P. antundseni n. sp., which has a stylet 17-19 mum long, is similar to P. tateae Wu &Townshend and P. labiosus Anderson &Kimpinski, but differs by the presence of males and the possession of conoid-truncate lip region, functional spermatheca, and long male tail (c = 8.5-9.5). Seriespinula seymouri Wu (Mehta &Raski), Nothocrieonema longulum (Gunhold) De Grisse &Loof, and Macroposthonia xenoplax (Raski) De Grisse &Loof are also reported from the islands. PMID- 19295716 TI - Post-infection Development and Morphology of Meloidogyne cruciani. AB - The development and life stages of Meloidogyne cruciani on tomato was studied at 28 C. Roots of 2-wk-old 'Rutgers' tomato seedlings were exposed to inoculum for 24 h, rinsed, and the seedlings repotted. No major changes in juvenile development were observed prior to 8 days after inoculation. At 11 days the second-stage juvenile had enlarged considerably. The genital primordium had not yet asumed the V-shape characteristic of developing females, but the presence of rectal glands identified the juveniles as females. At this time (11 days), two additional, previously undescribed esophageal lobes were first observed; they were adjacent to the dorsal and subventral glands. After molting from second to third stage, the stylet cone, shaft, and the lumen of the stylet knobs are shed and remain attached to the second-stage cuticle. The excretory duct of the third stage juveniles was directed anteriorly from the excretory pore of the second stage cuticle and appear attached to the body wall of the third-stage juveniles opposite the procorpus. At 19 days after inoculation, the last molt took place. The adult female possessed a new stylet, a large five-gland esophagus, a prominent excretory system ending in a unicellular gland and a fully developed reproductive system. PMID- 19295717 TI - Morphological Comparison of Head Shape and Stylet Morphology of Second-stage Juveniles of Meloidogyne Species. AB - Head shape and stylet morphology of second-stage juveniles of one population each of M. incognita, M. javanica, M. arenaria, and M. hapla were compared by light microscopy. Excised stylets of each species were also compared by scanning electron microscopy (SEM). Differences in head morphology were observed only between M. hapla and the other three species. In SEM, differences in stylet size, shape, and relative distance of the dorsal esophageal gland orifice to the base of the stylet were evident. Differences in stylet morphology between M. incognita and M. javanica could not he detected by light microscopy, but M. arenaria and M. hapla could be distinguished from each other and from the other two species. Head shape and styler morphology of second-stage juveniles are considered useful taxonomic characters. PMID- 19295718 TI - Influence of Soil Temperature on Meloidogyne incognita Resistant and Susceptible Cotton, Gossypium hirsutum. AB - The degree of resistance by a cotton plant to Meloidogyne incognita is affected by soil temperature, particularly in moderately resistant cultivars, The total number of nematodes in the resistant and moderately resistant rools at 35 C was equal to, or greater than, the number in susceptible roots at 20, 25, or 30 C. A shift in numbers to developing and egg-bearing forms of nematodes in the susceptible cultivar as tentperature increased indicates development was affected by temperature rather than by genetic resistance mechanisms. However, the nematode resistant cultivar did not support maturation of nematodes until a soil tempurature of 35 C was attained. This indicated that resistance mechanisms are partially repressed at 35 C and differences in nematode development cannot be explained in terms of accumulated heat units. The moderately resistant cultivar was significantly more sensitive to the effects of high temperature than was the resistant cultivar. PMID- 19295719 TI - Incidence and Distinguishing Characteristics of Meloidogyne chitwoodi and M. hapla in Potato from the Northwestern United States. AB - From September 1980 to June 1981, a survey was conducted in the major potato growing regions of northern California, Idaho, Nevada, Oregon. and Washington to determine the distribution of Meloidogyne chitwoodi and other Meloidogyne spp. Meloidogyne chitwoodi and M. hapla were the only root-knot nematode species detected parasitizing potato in all the states surveyed. Meloidogyne chitwoodi occurred alone in 83% of the samples and M. hapla in 11%, with 6% of all samples containing both species. The greater incidence of M. chitwoodi, as compared to M. hapla, may be due to the cool growing season encountered in 1980 (which favored M. chitwoodi but not M. hapla) and to the increased acreage of small grains (which are good hosts for M. chitwoodi but not M. hapla) planted in rotation with potato. Differentiation between these two species can be determined by a differential host test, perineal patterns of mature females, and shape of the tail tip amt of the tail hypodermal terminus of L juveniles. PMID- 19295720 TI - Influence of Initial Population Densities of Meloidogyne incognita on Three Chile Cultivars. AB - The effects of Meloidogyne incognita on the Big Jim, Jalapeno, and New Mexico No. 6 chile (Capsicum annuum) cultivars were investigated in microplots for two growing seasons. All three cultivars were susceptible to M. incognita and reacted similarly to different initial populations of this nematode. Severe stunting and yield suppressions occurred at all initial M. incognita densities tested ranging from 385 to 4,230 eggs and larvae/500 cm(3) soil. Regression analysis of the microplot data from a sandy loam soil showed yield losses of 31% for the 1978 season and 25% for the 1979 season for the three cultivars for each 10-fold increase in the initial population of M. incognita. PMID- 19295721 TI - Population Dynamics of Belonolaimus longicaudatus and Criconemella ornata and Growth Response of Bermudagrass and Overseeded Grasses on Golf Greens Following Treatment with Nematicides. AB - Portions of a 'Tifgreen' bermudagrass golf green with poor turf and large numbers of Belonolabnus longicaudatus and Criconemella ornata were treated with selected nematicides in the summers of 1977 and 1978. Improvements in turf quality were observed within 4 wk after treatment with phenamiphos and fensulfothion. Treatment with phenamiphos restulted in lower numbers of B. longicaudatus 4 and 14 wk after treatment in the 1977 experiment and up to 1 yr after treatment in the 1978 experiment. Treatment with fensulfothion reduced the number of B. longicaudatus for only 1 month after treatment and significantly increased the numbers of this nematode in September and March in the 1978 experiment, Negative correlations were obtained between numbers of B. longicaudatus and turf qualily up to 1 yr. Numbers of C. ornata were reduced only in January and June following treatment with phenamiphos and not at any time with fensulfothion. Treatment with fensulfothion resulted in higher numbers of this nematode than in check plots in November and March. The percent area covered by prostrate spurge the following year was reduced following treatment with phenamiphos, but not with fensulfothion. PMID- 19295722 TI - Suppression of Alfalfa Growth by Concommitant Populations of Pratylenchus penetrans and two Fusarium species. AB - Growth of alfalfa (Medicago sativa cv. Vernal) seedlings was compared after inoculation with combinations of either Pratylenchus penetrans and Fusarium soloni or P. penetrans and F. oxysporum f. sp. medicaginis. A synergistic disease interaction occurred in alfalfa when F. oxysporum and P. penetrans were added simultaneously to the soil. Alfalfa growth was suppressed at all inoculum levels of P. penetrans and F. oxysporum, but not with F. solani. Seedlings inoculated with the nematode alone gave lower yields than when inoculated with either Fusarium species alone. Fusarium oxysporum, but not F. solani, was pathogenic to alfalfa under similar experimental conditions. Fusarium oxysporum did not alter the populations of P. penetrans in alfalfa roots, whereas the presence of F. solani was associated with a diminished number of P. penetrans in the roots. PMID- 19295723 TI - Effects of Pratylenchus coffeae-Tylenchulus semipenetrans Interactions on Nematode Population Dynamics in Citrus. AB - The distributions of Pratylenchus coffeae and Tylenchulus semipenetrans in a central Florida citrus grove were mutually exclusive. In a challenge experiment conducted in the grove, indigenous populations of either species did not prevlude infection by the other species. Inoculation with either T. semipenetrans or P. coffeae tended to reduce the population size of the other nematode species. In greenhouse tests, individual feeder roots were parasitized predominantly by one or the other of the two species. Host response to parasitism in dual infections did not differ from response to single infection by either species. PMID- 19295724 TI - Biological Relationship of Rotyleinchulus borealis on Several Plant Cultivars. AB - The embryogenic development of Rolylenchulus borealis, at 24-26 C, was completed on corn, in 12-15 days, and the life-cycle of the nematode from egg to egg required 35-40 days at 20-25 C. Juveniles remained in the soil as preinfective stages for 17-19 days before becoming adults. Only immature vermiform and swollen egg-laying females were found attached to corn roots. Eggs were laid in a gelatinous matrix on the root surface; the number of eggs per egg mass was 45 +/- 28 on corn roots. Bean, green pea, potato, sorghum, and sweet potato were also found to be hosts of R. borealis. The nematode established a permanent feeding site on corn root in an endodermal cell that became hypertrophied. Pericyclic cells close to the feeding site showed granular cytoplasm and nuclei with hypertrophied nucleoli. A cell wall ingrowth was also noted around the area of stylet penetration into the endodermal cell. PMID- 19295725 TI - Cultivation of the Pine Wilt Nematode, Bursaphelenchus xylophilus, in Axenic Culture Media. AB - The pine wilt nematode, Bursaphelenchus xylophilus, has been cultured axenically in vitro on soy peptone/yeast extract or modified Caenorhabditis medium supplemented with cholesterol and hemoglobin. Although growth, development and reproduction were best in soy peptone/yeast extract medium, satisfactory population size increases were observed in the chemically defined Caenorhaditis medium. PMID- 19295726 TI - Decline and Death of Pinus spp. in Delaware Caused by Bursaphelenchus xylophilus. AB - Etiological studies to determine the cause of decline and death of Pinus spp. in Delaware were initiated in 1980. The pinewood nematode, Bursaphelenchus xylophilus, was found to be the major canse of mortality in Japanese black pine (Pinus thunbergii). When inoculated into healthy 5-yr-old Japanese black pines, B. xylophilus produced typical decline symptoms observed in the field. The xylophilous fungi most often associated with declining trees, Rhizosphaera pini, Fusarium spp., and Pestalotia funerea, were not pathogenic to Japanese black pine in greenhouse tests. Mineral analyses of soil and foliage showed no significant differences between healthy and infested trees. B. xylolyhilus was also found on loblolly pine (P. taeda), scrub pine (P. virginiana), Scots pine (P. sylvestris), red pine (P. resinosa), Eastern white pine (P. strobus), and pitch pine (P. rigida). PMID- 19295727 TI - Effect of Storage at Low Temperatures on Development and Survival of Postparasitic Juveniles of Romanomermis culicivorax (Nematoda: Mermithidae). AB - Development of postparasites and adult females of Romanomermis culicivorax Ross and Smith, 1976 may be retarded by keeping cultures at 10 or 15 C for 15 wk. Storage at 5 C resulted in high mortality of postparasites. A storage temperature of 10 C is suitable if development of postparasites, young adults, and gravid females is to be greatly retarded. None of the females were gravid when postparasites (1-6 days old) were kept at 10 C for 15 wk. Development resumed after the cultures were returned to 26 C and no significant mortality occurred. A storage temperature of 15 C is suitable if development is to be only moderately retarded. After a 15-wk storage period, only a few weeks were required to return the cultures to full production status. PMID- 19295728 TI - Infection of the Cat Flea, Ctenocephalides felis (Bouche) by Neoaplectana carpocapsae Weiser. AB - Infection of cat flea, Ctenocephalides felis, larvae by the entomophilic nematode Neoaplectana carpocapsae was accomplished in the laboratory. The Breton strain of N. carpocapsae provided higher larval mortality at lower dosages than did the DD 136 strain. Adult nematodes were evident in the insect hemocoel after 48 h; however, no infective third-stage larvae were produced. Larval flea infection increased with an increase in the moisture content of sand from 2% to 7% and of sandy clay from 7% to 12%. Larval flea infection was also obtained on turf containing dauer larvae. Nematode penetration of cocoons with invasion of prepupal and pupal fleas was apparent. PMID- 19295730 TI - Lectin Binding to Meloidogyne javanica Eggs. PMID- 19295729 TI - Influence of Host Nutrients on the Parasitic Development of Mermis nigrescens (Mermithidae). AB - Schistocerca gregaria nymphs and adults of both sexes were infected with eggs of Mermis nigrescens. Mermithid larvae grew more slowly in nymphal hosts, and emerging larvae were smaller than those from adult hosts. The longer the larvae remained in the host, the greater their size. Those developing in adult female hosts were longest. Single mermithid larvae that were transferred to a second host continued to grow and were significantly longer at emergence than larvae that developed solely in one host. In adult hosts that were infected with 40-300 M. nigrescens eggs, the percentage of mermithids that became males was strongly dependent on host weight at infective doses of 90 eggs or more. Results are discussed in relation to nutrient stress on the larvae and its importance in developing in vitro culture techniques. PMID- 19295731 TI - Detection and Ultrastructural Description of a Larval Moult in the Egg of Orrina phyllobia. PMID- 19295732 TI - Comparative Relationship between Meloidogyne chitwoodi and M. hapla population Densities and Growth of Sugarbeet Seedlings. PMID- 19295733 TI - Effects of the Temperature and Duration of the Initial Incubation Period on Resistance to Meloidogyne incognita in Tomato. PMID- 19295734 TI - Effect of Diurnal Changes in Soil Temperatures on Resistance to Metoidogyne incognita in Tomato. PMID- 19295735 TI - Effects of Heterodera fici on the Growth of Commercial Fig Seedings in Pots. PMID- 19295736 TI - Technique for Obtaining Eggs and Juveniles of Heterodera glycines. PMID- 19295737 TI - A technique for inoculating field sites with meloidogyne eggs. PMID- 19295738 TI - Technique for Gnotobiotic Cultivation of Heterodera glycines Ichinohe on Glycine max (L.) Merr. PMID- 19295739 TI - Growth and Development of Romanomermis culicivorax In Vitro. AB - Various combinations of vertebrate and invertebrate tissue culture and microbiological media were utilized in an attempt to culture Romanomermis culicivorax (Mermithidae: Nematoda) in vitro. Most media were unsuitable and caused nematodes to become lumpy, vacuolated, and granular. Slow and limited growth and development of internal structures of the nematodes were obtained with variously supplemented Grace's tissue culture and Schneider's Drosophila media. In an enriched Grace's medium, development attained by the nematodes after 3-4 wk was comparable to 4-5-day-old parasites grown in vivo in the mosquito host, Culex pipiens. Two molts were observed in vitro. Maximum dimensions in vitro were 7.0 mm length and 87-mum width at the widest point. The stichosome, stichocytes, and trophosome developed prominently. A filiform tail and highly cuticularized tube persisted throughout the culture period in vitro. PMID- 19295740 TI - Effects of Selected Insecticides and Nematicides on the In Vitro Development of the Entomogenous Nematode Neoaplectana carpocapsae. AB - The effects of organophosphates (mevinphos, phenamiphos, trichlorfon), carbamates (carbofuran, methomyl, oxamyl), a formamidine (chlordimeform), a synthetic pyrethroid (fenvalerate), a chlorinated hydrocarbon (methoxychlor). and an insect growth regulator (diflubenzuron) on in vitro development and reproduction of Neoaplectana carflocapsae were tested by incorporating each chemical into a nematode rearing medium. Organophosphates and carbamates adversely affected development and reproduction at concentrations >/= 0.1 mg/ml. Phenamiphos was the most toxic, with no nematode reproduction at 0.01 mg/ml. Inoculated infective juveniles developed to adults with some of the organophosphates and carbamates, but limited or no reproduction occurred. Chlordimeform inhibited development at 1.0 mg/ml, while diflubenzuron, fenvalerate, and methoxychlor did not significantly (P > 0.05) reduced reproduction at 1.0 mg/ml. The organophosphate and carbamate nematicides in use for control of plant-parasitic nematodes may be toxic to N. carpocapsae in the soil. PMID- 19295741 TI - Uptake of Lipids by the Entomophilic Nematode Romanomermis culicivorax. AB - Romanomermis culicivorax juveniles were dissected out of Aedes aegypti larvae 7 days after infection and incubated under controlled conditions in isotonic saline containing a (1)C-labeled fatty acid (palmitic acid), monoacylglycerol (glycerol monoolein), or triacylglycerol (glycerol tripalmate) nutrient source. The mermithid absorbed each of these lipids from the incubation medium, the rate of uptake being greatest for glycerol monoolein. No lipase activity was detected in whole nematode homogenates or in the media in which the nematodes were incubated. It is suggested that the nematode transports complex lipid molecules across its outer cuticle intact. PMID- 19295742 TI - Effect of alpha-Tocopherol and Culture Method on Reproduction of Turbatrix aceti. AB - The effect of alpha-tocopherol on the reproductive capacity of the free-living nematode Turbatrix aceti was determined using three different culture methods: mass culture, pair culture. and single culture. Significant differences were observed between control and alpha-tocopherol cultured nematodes for all reproductive parameters measured. The reproductive period started at a significantly earlier time and the length of the reproductive period was significantly longer in alpha-tocopherol cultured nematodes. The average number of offspring was 34 in control cultures as compared to 55 in alpha-tocopherol cultures. The eggs of alpha-tocopherol cultured females showed a more regular outline and uniform distribution of yolk than did eggs from control females. PMID- 19295743 TI - Description of Hoplolaimus magnistylus n. sp. (Nematoda: Hoplolaimidae). AB - Hoplolaimus magnistylus n. sp. is described and illustrated. It was found in soil around roots of soybean in Arkansas and Mississippi. It is similar to H. galeatus and H. concaudajuvencus. It differs from H. galeatus in all stages primarily by possession of a longer stylet. It differs from H. concaudajuvencus by the possession of rounded tails in second-stage juveniles vs. conically pointed tails with acute termini, having fewer subdivisions in female basal lip annules, and the greater distance from female anterior end to posterior end of esophageal lobes. Morphometrics and descriptions of second-, third-, and fourth-stage juveniles are given. A paratype female of H. sheri was examined and found to have six esophageal gland nuclei. PMID- 19295744 TI - Description of Dolichodorus grandaspicatus n. sp. (Nematoda: Dolichodoridae). AB - Dolichodorus grandaspicatus n. sp., collected from soil about roots of a red maple (Acer rubrum L.) in a creek bottom near Ludwig. Johnson County, Arkansas, is described. It differs from two closely related species, D. marylandicus and D. heterocephalus, by the longer spike on the female tail, the lenght of the female tail, and the shape of the extrudable portion of the gubernaculum of the males. It further differs from D. marylandicus by having heavily sclerotized accessory pieces of the vuvla and from D. heterocephalus by having a shorter stylet. PMID- 19295745 TI - Morphological Comparisons Between Xiphinema rivesi Daimasso and X. americanum Cobb Populations from the Eastern United States. AB - Though in the past Xiphinema americanum has been the most commonly reported dagger nematode in the eastern United States, our studies revealed the presence in Pennsvlvania of a previously unrecognized and unreported species related to X. americanum, Morphometric data and photomicrographs establish the identity of this form as X. rivesi and show expected variations in populations of this species from various locations. Similar data and illustrations are given for X. americanum populations from Pennsylvania and other areas, showing variations and relationships. Xiphinema rivesi is widely distributed in the fruit producing area of south-central Pennsylvania and is also reported herein from raspberry in Vermont and apple in Maryland and New York. This species is frequently found in fruit growing areas of Pennsylvania associated with tomato ringspot virus-induced diseases and is also found associated with corn, bluegrass sod, and alfalfa. PMID- 19295746 TI - Simulated sampling strategies for nematodes distributed according to a negative binomial model. AB - A FORTRAN computer program was developed to simulate nematode soil sampling strategies consisting of various numbers of samples per field, with each sample consisting of various numbers of soil cores. The program assumes that the nematode species involved fit a negative binomial distribution. Required input data are estimates of the mean and k values, the number of samples per field and cores per sample in the strategy to be investigated, and the number of times the simulation is to be replicated. Output consists of simulated values of the relative deviation from the mean and standard error to mean ratio, both averaged over all replications. The program was used to compare 150 simulated sampling strategies for Meloidogyne incognita, involving all combinations of two mean values (2.0 and 10.0 la.rvae/10 cm(3) soil), three k values (1.35, 0.544, and 0.294), five different numbers of samples per field (1, 2, 4. 10, 20), and five different numbers of cores per sample (1, 2, 4, 10, 20). Simulations resulting from different mean values were similar, but best results were obtained with higher k values and 20 cores per sample. Relatively few 20-core samples were needed to obtain average deviations from the mean of 20-25%. PMID- 19295747 TI - Estimating relative error in nematode numbers from single soil samples composed of multiple cores. AB - Spatial distributions of several species of plant-parasitic nematodes were determined in each of three fallow vegetable fields and in smaller subunits of those fields. Goodness of fit to each of several theoretical distributions was tested hy means of a X(2) test. Distributions for most species showed good agreement with a negative binomial model. An exception occurred with Crictmemella sp., which showed a better fit to the Neyman Type A distribution. For nematodes distributed according to the negative binomial model, the number of cores per composite sample needed to achieve specified relative errors was calculated. For a given nematode species, such as Quinisulcius actus (Allen) Siddiqi or Meloidogyne incognita (Kofoid &White) Chitwood, the k values for the negative binomial distribution increased as field size decreased, with the result that fewer cores were needed to achieve the same level of precision in a smaller field. Best results were achieved when the single sample was used to estimate populations in fields of 0.25-0.45 ha in size. When using only a single composite sample to estimate mixed populations of the nematodes studied here in a field of that size, approximately 22 cores per composite sample would be needed to estimate all population means within a standard error to mean ratio of 25%. Considerably, more cores were needed to maintain a given level of precision in fields of 1.0 ha or greater, and it may be necessary to subdivide larger unils (ca. 1.5 ha and up) for accurate sampling. PMID- 19295748 TI - Influence of Potassium and Nitrogen Fertilization on Parasitism by the Root-knot Nematode Meloidogyne javanica. AB - The influence of various c oncentrations of K, nitrogen sources, and inoculation with root-knot nematode Meloidogyne javanica were evaluated in tomato plants. Increased potassium concentration increased top and root fresh weights of intact plants and fresh weights of excised roots. Nitrate-fertilized plants weighed more than plants receiving ammonium independent of the K level in the medium. Nematode counts on roots were not affected by nutritional differences in intact or excised roots. In intact roots a high percentage of males was recorded at low K levels, whereas in excised roots the proportion of males in the population rose as the K levels increased. Inoculated intact roots accumulated K when the level of potassium supply was low; infected excised roots contained less K than did nematode-free roots. PMID- 19295749 TI - Optimum Initial Inoculum Levels for Evaluation of Resistance in Tomato to Meloidogyne spp. at Two Different Soil Temperatures. AB - The effects of Meloidogyne incognita or M. javanica at five initial inoculum levels of 20, 100, 200, 1,000, and 2,000 eggs and infective juveniles per seedling on 'Floradade,' 'Nemarex,' 'Patriot,' and 'PI 129149-2(sib)-5' tomatoes maintained at 25 or 32.5 C were studied. The number of egg masses on roots of the susceptible cultivar Floradade was similar for both species of root-knot nematodes at either 2.5 or 32.5 C soil temperatures. At 25 C, very low numbers of egg masses were produced by both species of root-knot nematodes on Nematex, Patriot, and Lycopersicon peruvianum PI 129149-2(sib)-5. At 32.5 C, the best inoculum level for assessing resistance in these tomato genotypes was 200 eggs and infective juveniles per seedling. With 28 days of incubation, this temperature and inoculum level produced quantitative differences in resistance for both species of Meloidogyne. PMID- 19295750 TI - Genetic Basis of the Epidemiologic effects of Resistance to Meloidogyne incognita in the Tomato Cultivar Small Fry. AB - The genetic nature of resistance and its epidemiologic effects on two Meloidogyne incognita populations were assessed in the F hybrid tomato cv. Small Fry. The progeny of a Small Fry x Small Fry cross segregated in a 3:1 resistant:susceptible ratio, indicating the presence of a single, completely dominant resistance gene (LMiR) in Small Fry. In a subsequent experiment, infection frequency and the rate of development of primary infection on resistant Small Fry x Small Fry segregates were compared to those on susceptible segregates and the susceptible cultivar Rutgers. Suppression in both infection frequency and rate of development of primary infection was entirely attributable to gene LMiR. A single egg-mass population of M. incognita propagated for 12 generations on Small Fry showed an increased ability over the wild type population to parasitize plants containing the LMiR gene but failed to completely overcome resistance. The relationship of this phenomenon to the genetics of the Lycopersicon esculentum-M. incognita interaction is discussed. PMID- 19295751 TI - Development of Four Populations of Meloidogyne hapla on Two Cultivars, of Cucumber at Different Temperatures. AB - The infectivity and development of four populations of Meloidogyne hapla were compared, at three temperatures, on tomato and two varieties of cucumber. A population from Canada produced few root-galls on cucumber and, except at 24 C, no larvae developed into adult females and produced egg masses. In contrast, a population with 45 chromosomes from America produced many galls on cucumber and small proportions of larvae became females and produced egg masses at 20 and 24 C. At 18 C this population produced no egg masses on cucumber, but a population from Britain and one from America with 17 chromosomes produced more egg masses at this temperature than at 20 or 24 C. Dissection of the galls showed that on cucumber many larvae died or their growth and development was slowed. PMID- 19295752 TI - Sex Differentiation in Meloidogyne incognita and Anatomical Evidence of Sex Reversal. AB - Sex differentiation was studied by examining the cellular structure of gonad primordia extracted from second-stage juveniles developing under different environmental conditions. In female jnveniles, divisions of the two somatic cells of the primordium occurred in mid-sccond stage and resulted in 12 cells. Two of them were differentiated as cap cells, two occupied the anterior central and eight the posterior central part of the V-shaped primordium. The two germinal cells divided at the 6-8 somatic-cell stage of the primordium; i.e., earlier than in any other plant-parasitic nematode. In male juveniles of similar developmental stage, divisions of somatic cells resulted in 10 cells: one cap cell at the posterior tip and nine cells at the anterior part of the rod-shaped primordium. Germinal cells divided at the 6-8 sontatic-cell stage. On the basis of gonad anatomy it was concluded that some female juveniles undergo sex reversal and proceed with further development as males. The degree of expression of intersexual features depends on the period at which sex reversal occurs. Sex reversal at an early period gives rise to males with one testis, almost indistinguishable front true males. Sex reversal at mid-second stage involves degeneration of the nucleus of one of the cap cells resulting in males with an atrophied testis and a well-developed testis. More delayed sex reversal results in males with two testes of approximately equal size. To explain these patterns of development, it is assumed that sex differentiation is hormonally controlled and that the environment influences hormonal balance by affecting gene expression. PMID- 19295753 TI - Effect of Planting Site Preparation, Hydrated Lime, and DBCP (1, 2-dibromo-3 chloropropane) on Populations of Macroposthonia xenoplax and Peach Tree Short Life in Georgia. AB - Annual postplanting applications of 40.7 kg/ha DBCP (1,2-dibromo-3-chloropropane) controlled Macroposthonia xenoplax (Raski, 1952) deGrisse and Loof, 1965 in peach tree short life sites, reduced bacterial canker incidence from 74% to 6%, and inreased the average life of the trees from 3.9 to 6.8 yr for a 7-yr test period. Hydrated lime at 5.5 kg per planting site reduced bacterial canker incidence from 81% 57 % and increased the tree longevity from 2.6 to 6.0 yr. Populations M. xenoplax were inversely correlated with tree longevity. PMID- 19295754 TI - Population Dynamics of Criconemoides simile on Soybean. AB - Custer and Hood soybean cultivars were inoculated with nine levels of Criconemoides simile ranging from 300 to 20,600 nematodes per plant. Rate of reproduction decreased as inoculum level was increased beyond 900-2,000 nematodes. Final population density was influenced by both composition and level of inoculum. There was an indication that substance(s). inhibitory to larvae, accumnlated in soil in which Hood was grown for 11 months, Significant reduction of fresh weight of roots of Hood, but not Custer, occurred at population densities of 37,000 and 44,700 nematodes per plant. PMID- 19295755 TI - Response of Peanut, Corn, Tobacco, and Soybean to Criconemella ornate. AB - The relative susceptibility of four field crops to Criconemella ornata differed greatly in microplot tests. As few as 178 freshly-introduced C. ornata/500 cm(3) of soil stunted peanut. In contrast, this nematode had no effect on the growth of corn or soybean. Large populations remaining after culture of peanut or corn enhanced the growth of tobacco. A problem of comparing the effects of a freshly introduced population of this nematode with large residual populations was encountered. Freshly extracted, greenhouse-grown inoculum caused the typical "yellows disease" on peanut, whereas much greater residual population densities following a poor host (tobacco) had little effect on the growth of peanut. It is suggested that many of the nematodes in the field following a poor host are dead. Peanut supported greater reproduction (up to 970-fold) than did other crops tested. Corn was intermediate, with a population increase as great as 264-fold; soybean and tobacco failed to maintain initial population densities. PMID- 19295756 TI - Estimate of yield loss from the citrus nematode in Texas grapefruit. AB - Chemical control of the citrus nematode, Tylenchulus semipenetrans Cobb, has consistently increased yield of grapefruit on sour orange rootstock in Texas. In this study, data from chemical control tests conducted from 1973 to 1980 were analyzed to determine the relationship between nematode counts and grapefruit yield and fruit size. The correlation between yield and nematode counts was negative (r = -0.47) and highly significant (P < 0.01). The data best fit the exponential decay curve: y = 160.3e(-0.0000429) where y = yield in kg/tree and x = nematodes/100 cm(3) of soil. The correlation between fruit size and nematode counts was not significant because yield and fruit size were inversely related. Yield loss in an average untreated orchard was estimated to be 12.4 tons/ha. Economic loss to citrus nematode in Texas grapefruit, assuming no treatment and an average on-tree price of $60/ton, was estimated to be $13.2 million annually. PMID- 19295757 TI - Effect of Tylenchorhynchus robustoides on Growth of Buffalo Grass and Western Wheatgrass. AB - Tylenchorhynchus robustoides reduced (P = 0.05) growth of Agropyron smithii (western wheatgrass) at soil temperatures of 20, 25, 30, and 35 C. Growth reduction increased with increasing soil temperatures. Highest populations of T. robustoides were recovered at 25 and 30 C. Clipping weights of Buchloe dactyloides (buffalo grass) were reduced at 25 and 30 C; however, root/crown weights were reduced at 15, 20, 30, and 35 C in nematode infested vs. noninfested soil. Reproduction of T. robustoides was greater at 25, 30, and 35 C than at 20 C on B. dactyloides. In a greenhouse study, T. robustoides reduced clipping and root/crown weights of both grasses 24-64%. PMID- 19295758 TI - A Comparison of the Hatching of Juveniles from Cysts of Heterodera schachtii and H. trifolii. AB - The effects of root diffusates of selected plants within the families Chenopodiaceae and Cruciferae and the hatching agent zinc chloride were tested for their effects on hatching and emergence of juveniles from cysts of Heterodera schachtii and a race of H. trifolii parasitic on Chenopodaceae and Cruciferae in The Netherlands. Although all diffusates strongly stimulated hatching of juveniles of H. schachtii, their effects on H. trifolii were less evident. PMID- 19295759 TI - Screening Soybean for Resistance to Heterodera glycines Ichinohe Using Monoxenic Cultures. AB - A simple, rapid, and inexpensive method for evaluation of host-parasite interactions, based on monoxenic cultures, is described. Axenic root explants of Glycine max (L.) Merr., cultured on a holidic agar medium, were inoculated with axenic second-stage larvae of Heterodera glycines Ichinohe, Race 3. A clear separation of susceptible and resistant cultivars, based on numbers of mature female nematodes present after 3 wk at 25 C, was observed. The method described should aid researchers in the evaluation of the host response to infection by H. glycines. PMID- 19295760 TI - Evaluation of Neoaplectana carpocapsae for Biological Control of the Western Spruce Budworm, Choristoneura occidentalis: Ineffectiveness and Persistence of Tank Mixes. PMID- 19295762 TI - Phoresy Between a Mushroom-infesting Fly and Two Free-living Nematodes Associated with Mushroom Culture. PMID- 19295761 TI - Race Comparisons of Heterodera glycines Using Crossed Immunoelectrophoresis. PMID- 19295763 TI - Effect of Temperature on Reproduction and Motility of Pratylenchus vulnus. PMID- 19295764 TI - An illustrated key to the cyst-forming genera and species of heteroderidae in the Western hemisphere with species morphometrics and distribution. AB - Diagnoses of the cyst-forming genera of Heteroderidae (viz., Heterodera, Sarisodera, Globodera, Punctodera, Cactodera, and Dolichodera) and distribution and morphometrics of the 34 known cyst species in the Western Hemisphere are presented along with an illustrated key for the identification of these genera and species. The key is based mainly on cysts and larvae, and important morphological and diagnostic features are extensively shown by LM and SEM illustrations. The genus Bidera is placed as a new synonym under the genus Heterodera. PMID- 19295765 TI - Redescription of Heterodea zeae, the Corn Cyst Nematode, with SEM Observations. AB - Heterodera zeae, the corn cyst nematode, is redescribed and illustrated with comparative details and measurements of females, cysts, and larvae from Maryland, USA; and India. Scanning electron micrographs o f specimens from the United States are also presented. Revised measurements for the larval stylet and new diadnostic characters, especially in the cyst cone, for H. zeae are given. The relationship of H. zeae to close species is discussed. PMID- 19295766 TI - Description of Distolabrellus veechi n. gen., n. sp. (Nematoda:Rhabditidae). AB - A new, monotypic genus and species in the subfamily Mesorhabditinae is described and illustrated. The primary character which separates the species and genus from all others in the Rhabditidae is a lip region with six set-off and well-separated liplets of two alternating shapes. Other diagnostic characters are transverse and fine longitudinal body striations, cuticular micropores, a prismatic prostom, well-developed esophageal collar, and three denticles on each metarhabdion. Males have a peloderan, open bursa bearing two pre- and seven post-anal bursal papillae, fused spicules over 50 mum long, and paired diverticulate cement glands associated with the vas deferens. PMID- 19295767 TI - Description of Paktylenchus tuberosus gen. n., sp. n. (Nematoda: Tylenchinae). AB - Paktylenchus tuberosus gen.n., sp.n. is described and illustrated from soil around Solanum tuberosum from Kaghan Valley, Pakistan. Paktylenchus gen.n, can be distinguished from the most closely related genus, Agelenchus Andrassy, 1954, by the distinct longitudinal striae, three incisures in the lateral field, and absence of males. Paktylenchus tuberosus shows affinities to Coslenchus Siddiqi, 1978 but differs in having an oblique vagina-to-body axis and a prominent sunken vulva with vulval flaps. Four genera of the subfamily Tylenchinae are also discussed herein. PMID- 19295768 TI - Relationship Between Paratrichadorus sp. Density, and Growth of Wheat in Pots. AB - The effect of a Paratrichodorus sp. (close to P. tunisiensis) on the growth of wheat (Triticum durum Desf.) was investigated in pots containing different nematode densities and maintained in a growth chamber at 20 C for 40 days. The relation between fresh weight of tops and initial nematode density was according to the equation y = m + (1 - m)z(PT). This suggests a tolerance limit of 1.4 nematodes/cm(3) of soil under the conditions of the experiment; taking into account the effect of the great nematode mortality, it is estimated to be between 0.15 and 0.35 nematodes/cm(3) soil. Models of the growth of the plants and the multiplication of the nematodes (assuming a constant mortality of the nematodes in the absence of roots) which explain the relation between initial and tinal nematodes densities at initial densities greater than 1 nematode/cm(3) soil are described in an appendix. Sections of nematode infested roots showed disorganization of root structure clue to abnormal proliferation of lateral roots. Nematode feeding on the root cap and apical meristem caused cessation of root elongation and induced abnormal production of lateral root primordia. PMID- 19295769 TI - Reproduction of Criconemoides simile, Helicotylenchus pseudorobustus, and Paratylenchus projectus on Soybean. AB - Reproduction of Criconernoides simile, Paratylenchus projectus, and Helicotylenchus pseudorobustus, alone and in certain combinations, was characterized during 50 days on the soybean cultivars Custer and Hood. Custer was the more suitable host for all three nematodes and C. simile was the most vigorous parasite on both cultivars. C. simile and H. pseudorobustus reproduced as well in combination as they did alone; addition of P. projectus to this combination did not alter results. Reproduction by P. projectus was repressed on both cultivars when it was combined with C. simile and H. pseudorobustus. The relative importance of time was assessed, as it relates both to the establishment of parasitism by applied nematodes and to the rate of development of larvae in the various host-parasite combinations. The necessity of characterizing as well as enumerating populations when evaluating population development of ectoparasitic nematodes was demonstrated. PMID- 19295770 TI - Efficacy of Selected Nonvolatile Nematicides on Control of Ditylenchus destructor in Iris. AB - Greenhouse and field tests established that fenamiphos at 6.7 and 13.4 kg ai/ha applied in a 30-cm band directly on iris bulbs at planting effectively controlled Ditylenchus destructor. Aldicarb at rates of 5.6 to 11.2 kg ai/ha was less effective. Carbofuran, fensulfothion, and oxamyl at 6.7 to 13.4 kg ai/ha were ineffective. When applied on the bulbs, fenamiphos (granular or liquid) reduced nematode infection from 31 to 0.6% as determined by visual inspection of bulbs at harvest. Populations of D. destructor were reduced from 5.7 nematodes/g of fresh weight of bulb tissue to 0.04, 0.05, and 0.14 with applications of 13.4, 6.7, and 3.3 kg ai/ha fenamiphos, respectively. The most effective treatment was fenamiphos (granular or liquid) applied in a 30-cm band directly on the bulbs at time of planting. PMID- 19295771 TI - Effect of Time of Application on the Action of Foliar Sprays of Oxamyl on Meloidogyne hapla in Tomato. AB - Foliar sprays containing 3,000 or 4,000 ppm oxamyl applied before inoculation with Meloidogyne hapla completely protected tomato plants from intection for up to 36 days but sprays containing 1,000 or 2,000 ppm provided only partial protection. Postinoculation sprays were less effective than preinoculation sprays but they decreased the numbers of females and their rate of development and increased the numbers of males. Similar amounts of oxamyl applied to the soil as a drench or as granules controlled M. hapla more effectively than foliar sprays but the longer treatment was delayed after infection the fewer the larvae that were killed and the more that became male. PMID- 19295772 TI - Development of Meloidogyne incognita Inhibited by Digtaria decumbens cv. Pangola. AB - Population densities of the root-knot nematode, Meloidogyne incognita, were lower after 90 days in soil planted to digitgrass (Digitaria decumbens cv. Pangola) than in soil left fallow or planted to tomato. Roots of tomato seedlings interplanted with Pangola digitgrass were less galled than were roots of tomato seedlings planted alone. Fewer second stage larvae invaded roots of Pangola digitgrass than tomato and those that entered the grass roots failed to develop beyond the late second stage. PMID- 19295773 TI - Population development and reproduction of fungus- and bacterium feeding nematodes in the presence of insect growth regulators. AB - The insect growth regulators (IGRs), diflubenzuron and BAY SIR 8514, at 300 and 1,000 ppm a.i. in potato dextrose agar (PDA) inhibited the radial growth of the fungus Rhizoctonia solani host of Aphelenchus avenae. The IGRs had no effect on the growth of the bacterium Pseudomonas pseudoalcaligenes host of Acrobeloides nanus and Diplogaster iheritieri. At 59 ppm a.i., neither IGR inhibited the population development of A. nanus and D. iheritieri on P. pseudoalcaligenes; however, diflubenzuron stimulated the population development of D. iheritieri. At 300 ppm, both IGRs inhibited the population development of A. nanus and D. iheritieri; however, BAY SIR 8514 was more effective than diflubenzuron except on A. nanus L's. At 300 ppm, only BAY SIR 8514 affected the population development of A. avenae, except the L's. At 1,000 ppm, both IGRs inhibited development, except diflubenzuron for L and L's. Again, BAY SIR 8514 was more effective than diflubenzuron. With single females of A. nanus and D. iheritieri, both IGRs at 300 ppm reduced egg laying, inhibited embryonation, and slowed larval development. PMID- 19295774 TI - Effect of Soil Water Potential on Survival of Meloidogyne javanica in Fallow Soil. AB - A natural infestation of Meloidogyne javanica in an aggregated Oxisol declined at an exponential rate when aliquots of the soil were stored for 72 days in polyethylene bags at various soil water potentials (Psi). Time periods required for reduction in soil infestations by 50% were 2.7, 4.9, 110, 10, and 2.6 days at Psi of -0.16, -0.30, -1.1, -15, and -92 bars, respectively. In the wetter soils, at Psi of -0.16, -0.30, and -1.1 bars, the predominant stage recovered was the second-stage larva. In the drier soils, at Psi of -15 and -92 bars, both eggs and larvae were recovered with neither stage predominating. Incidence of coiled larvae was inversely related to the Psi value of the soil, a greater incidence occurring in the drier soils. After 15-32 days, percentages of coiled larvae were 13, 27, 55, 65, and 88% in soil at Psi of -0.17, -0.60, -1.9, -15, and -82 bars, respectively. PMID- 19295775 TI - Life Cycle of Heterodera zeae Koshy, Swarup, and Sethi on Zea mays L. Axenic Root Explants. AB - Monoxenic cultures of Heterodera zeae, the corn cyst nematode (CCN), were established on root explants of corn Zea mays L., cv. Kenworthy. The life cycle of H. zeae was determined from light anti scanning electron microscopic observations of the root explants grown in the dark at 29.5 +/- .5 C under gnotobiotic conditions. The life cycle, from the time the explants were inoculated with second-stage larvae (L2) to the first appearance of newly hatched second-generation L2, required 22 days. The occurrence of males was rare suggesting that reproduction in H. zeae is parthenogenetic. PMID- 19295776 TI - Response of Soybean Lines to Differentially Selected Cultures of Soybean Cyst Nematode Heterodera glycines Ichinohe. AB - Eight soybean cyst nematode, Heterodera glycines Ichinohe, cultures were developed by their continuous selection on soybean lines with various level of resistance. Each soybean line was inoculated with eggs and larvae of the eight cultures. Female development was much higher on 'Essex' and 'Forrest' than the other lines. The development of nematode cultures selected on Essex and Forrest was low on the other six soybean lines compared with the lines on which they were selected. PI 89772 and PI 90763 had a high level of resistance to the culture selected on PI 88788, whereas cultures selected on PI 89772 and PI 90763 reproduced very little on PI 88788 and PI 209352. Cultures selected on PI 87631 1, PI 209332, and 'Cloud' gave response similar to that of PI 88788. The use of various sources for resistant variety development program is discussed. PMID- 19295777 TI - Effects of Temperature and Root Leachates on Embryogenic Development and Hatching of Meloidogyne chitwoodi and M. hapla. AB - At 20 C the duration of the embryogenic development of Meloiclogyne chitwoodi and M. hapla was about 20 days. At 10 C the embryogenic development was 82-84 days for M. chitwoodi and 95-97 days for M. hapla. The effect of distilled water and root leachates of potato cv. Russet Burbank, tomato cv. Columbian, and wheat cv. Hyslop on the hatching of eggs of the two root-knot nematode species was investigated at 4, 7, 10, 15, 20, and 25 C (+/- 1 C). Cumulative egg taatch was no greater in root leachates titan in distilled water, but temperature did significantly affect egg hatch (P = 0.05). Less than 1% of the eggs of both nematode species hatched at 4 C. The percent cumulative hatch at 10 C was significantly less (P = 0.05) than at higher temperatures for both nematodes and significantly more (P = 0.05) M. chitwoodi eggs hatched than did M. hapla eggs. At 15 G the percent cumulative hatch of both species was significantly lower (P = 0.05) than that at 20 and 25 C. The percent cumulative egg hatch of two species did not differ at 25 C, but was higher (P = 0.05) at 25 C than at 20 C. At 7 C the emergence of M. chitwoodi juveniles was about seven times (P = 0.01) greater than that of M. hapla in distilled water. PMID- 19295778 TI - Ultrastructural Changes Caused by Fusarium oxysporum f. sp. lycopersici in Meloidogyne javanica Induced Giant Cells in Fusarium Resistant and Susceptible Tomato Cultivars. AB - Tomato (Lycopersicon esculentum Mill.) seedlings, susceptible (cv. Pearson A-I Improved) and resistant (cv. Pearson Improved) to race 1 Fusarium oxysporum f. sp. lycopersici (Sacc.) Snyd &Hans., were inoculated with Meloidogyne javanica (Trueb) Chitwood second-stage juveniles and 3 weeks later with race 1 F. oxysporum f. sp. lycopersici spores. One week after fungal inoculation, no fungus was visible in root tissue of the tomato cultivars and the giant cells were normal. Two weeks after fungal inoculation, abundant hyphae were visible in xylem tissues of Fusarium-susceptible but not of Fusarium-resistant plants. In susceptible plants, giant cell degeneration occurred, characterized by membrane and organelle disruption. In addition, where hyphae were in direct contact with the giant cell, dissolution of the giant cell wall occurred. Three weeks after fungal inoculation, fungal hyphae and spores were visible inside xylem tissues and giant cells in Fusarium-susceptible plants and in xylem tissue of the resistant plants. In susceptible and resistant plants, giant cell degeneration was apparent. Giant cell walls were completely broken down in Fusarium susceptible tomato plants. In both cultivars infected by Fusarium, giant cell nuclei became spherical and dark inclusions occurred within the chromatin material which condensed adjacent to the fragmented nuclear membrane. No such ultrastructural changes were seen in the giant cells of control plants inoculated with nematode alone. Giant cell deterioration in both cultivars is probably caused by toxic fungal metabolites. PMID- 19295779 TI - Host Suitability and Susceptibility of Glycine max cv. Bedford to Race 1 of Heterodera glycines. AB - Populations of Heterodera glycines identifiable as race 1 reproduced on the race 1 resistant 'Bedford' soybean. A Beaufort County, North Carolina, population had an index of parasitism of 112% on Bedford in greenhouse tests. Indices of parasitism for this population on race 1 resistant cultivars Pickett 71, Centennial, and Forrest were less than 10%. The Beaufort County population had significantly greater reproduction on Bedford in microplots than did populations of race 3 or race 4. In field tests, a race 1 population suppressed yields of Bedford but not yields of Centennial. Based on these data, Bedford is no longer recommended in North Carolina as a race 1 resistant cultivar. PMID- 19295780 TI - Comparative Response to Sugarbeet and Fodder Beet to Heterodera schachtii. PMID- 19295781 TI - An improved technique for clearing and staining plant tissues for detection of nematodes. PMID- 19295782 TI - Reaction of Beta spp. to Root-knot Nematodes. PMID- 19295783 TI - Observations on the Inner Labial Sensilla of Second-stage Larvae of Heterodera glycines. PMID- 19295784 TI - Gall Formation on Pistacia vera by Rotylenchulus macrodoratur. PMID- 19295785 TI - Nematode chemosensilla: form and function. AB - As an introduction to a symposium of nematode chemoreception, the anatomy of nematode chemosensilla, their distribution on plant parasitic nematodes, and their possible functional roles is briefly reviewed. Comparison of nematode chemosensilla with those of other animals shows their greater resemblance to olfactory primary sense cells of vertebrates. Although the sensory process is obviously derived from a cilium, the absence of many ciliary features is noted. Retention of the ciliary necklace may be important functionally. A simple model is proposed, wherein binding of stimulant molecules to receptors in the membrane of the cilium-derived process results in entry of Na and Ca (the latter via the ciliary necklace) to produce a receptor potential that spreads along the dendrite to the cell body where action potentials continue along the short axon to synapses. PMID- 19295786 TI - Biochemical studies of olfaction: role of cilia in odorant recognition. AB - Chemoreception in vertebrates is beginning to be understood. Numerous anatomical, behavioral, and physiological studies are now available. Current research efforts are examining the molecular basis of chemoreception. Rainbow trout (Salmo gairdneri) have a functional olfactory system and are a suitable vertebrate model for studying odorant interactions with receptors. Using a biochemical approach, initial events of olfactory recognition were examined; the aim was to determine the location and specificity of odor receptors. Cilia occupy the distal region of the receptor neuron on the trout olfactory epithelium, and their membranes are the postulated locus of odorant receptor sites. A cilia preparation was isolated from the olfactory rosette. The preparation was characterized by quantifying biochemical markers for cilia, along with electron microscopy, all of which substantiated enrichment of cilia. Functional activity was assessed by quantifying binding of several radioactively labeled odorant amino acids. The odorants bound to the cilia in a manner similar to the sedimentable preparation previously isolated from t h e olfactory rosette of the same animal, thus verifying the presence of odor receptors in the cilia preparation. Evidence also confirmed a site TSA which binds L-threonine, L-serine, and L-alanine and a site L which binds L-lysine (and L-arginine). Binding of L-serine and D-alanine showed evidence for a single affinity site while the others showed two affinity sites. Separation of membrane fractions from the cilia preparation revealed that binding activity is associated with a very low density membrane fraction B. PMID- 19295788 TI - Hypotheses and possibilities of intervention in nematode chemoresponses. PMID- 19295787 TI - Chemotactic behavior of nematodes. AB - Nematode chemoreception is reviewed Methods that have been used to measure chemotaxis are discussed and a new method using a video camera interfaced to a microcomputer is briefly described. The chemical stimuli that have been identified are discussed. The transition from attractant to repellent as NaCl increases in concentration is demonstrated by new data. PMID- 19295789 TI - Cryphodera utahensis n. sp., (Heteroderidae), A New Species from Wild Rose in Utah. AB - A new species of Cryphodera Colbran, 1966, parasitic on wild rose (Rosa sp. L.), is described from Utah, USA. Cryphodera utahensis n. sp. most closely resembles C. podocarpi (Wouts, 1973) Luc et al., 1978, but is distinct with respect to a number of characters including juveniles with a longer esophagus (206 versus 142 mum), shorter tail (47 versus 69 mun), and four rather than three incisures in the lateral field. Females of C. utahensis have a terminal protuberance which is more pronounced than in other species. Scanning electron microscope observations of the lip region of males indicate a labial disc surrounded by six lip sectors. However, the remainder of the lip region is comprised of irregularly-shaped plates; similar patterns with longitudinal striae apparently characterize males throughout the genus. The lip pattern of juveniles of C. utahensis n. sp. includes an oval labial disc surrounded by six lip sectors; transverse striae extend the height of the lip region without longitudinal striae. The type locality of C. utahensis confirms a broader distribution of the genus than was indicated for other species, which are limited to Australia (Queensland) and New Zealand. PMID- 19295790 TI - Descriptions of Chitwoodius brasiliensis n.sp., Chitwoodius rusticulus n.sp., and Vancterlindia venata n.sp. (Dorylaimida: Tylencholaimidae: Vanderlindiinae). AB - Chitwoodius brasiliensis n.sp. is described from soil around plant roots in Vicosa, MG State, Brazil. It has a body 1.9 mm (1.6-2.5) long, odontostyle and odontophore 33 mum (31-37) and 34 mum (31-36) long, respectively, and males with spicules 59-61 mum long. Chitwoodius rusticulus n.sp. from Colombian rain forest differs from other species of the genus in having a weakly muscular anterior part of the oesophagus, a pore-like vulva, and an unsclerotized vagina. Vanderlindia venata n.sp. from lucerne soil in South Kilimanjaro, Tanzania, differs from the type and only other known species of its genus in having a smaller and less slender body (L = 3.34 mm [3.15-3.71], a = 50 [46-53]), odontostyle 2.3-2.5 times lip region width long, and stylet guiding ring located at 1.2-1.4 times lip region width from anterior end. PMID- 19295791 TI - Effect and Reproduction of Rotylenchulus reniformis on Sweet Potato Selections. AB - Growth, yield, and quality of 10 sweet potato selections and reproduction of the reniform nematode, Rotylenchulus reniformis, were studied in fumigated and nonfumigated plots in a naturally infested field. Nematode reproduction on the different selections in the field was similar to that reported in the greenhouse but was not related to the effect of the nematode on yield of the different selections. Goldrush supported the least reproduction but was the most severely affected by the nematode, while Centennial supported the most reproduction but was the least affected. Although reniform nematode was not found within enlarged fleshy roots, sweet potatoes were more frequently cracked in nonfumigated plots even when nematode populations were relatively low. One selection, P-104, was resistant to cracking. Yield of all selections tested was significantly reduced when initial populations were moderate to high (1,500-10,000 nemas per 500 cm(3)). Correlations were made between nematode population parameters and growth, yield, and cracking of the sweet potatoes. The initial populations and the reproduction ratio for the last part of the growing season gave the most significant negative correlations with yield for most selections. PMID- 19295792 TI - Population Dynamics of Meloidogyne incognita and Rotylenchulus reniformis Alone and in Combination, and Their Effects on Sweet Potato. AB - Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) interactions on sweet potato were studied in naturally and artificially infested field plots for 3 years. In a naturally infested field, early season counts of Mi or Rr were positively correlated with later counts of the same nematode, but negative correlations were found between early Mi and subsequent Rr, and early Rr and subsequent Mi counts. In field plots fumigated with methyl bromide and then infested with low levels of Rr, Mi, and Rr + Mi, final population densities of Mi juveniles were reduced by Rr, but Rr was not affected by Mi. In field plots with a high natural population density of Rr, artificial infestation with high levels of Mi in both fumigated and nonfumigated treatments inhibited Rr, while the final Mi juvenile population density was not affected. Results indicate that a competitive interaction exists with each species capable of inhibiting the other and becoming the dominant population. The nematodes had no apparent effect on yield at the inoculum densities used, either alone or mixed. Both nematodes increased cracking of sweet potatoes, but mixed populations did not differ in incidence of cracking from either Rr or Mi alone. PMID- 19295793 TI - Parasitism of Rotylenchulus reniformis on Soybean Root Rhizobium Nodules in Venezuela. PMID- 19295794 TI - Effects of Concomitant Development on Reproduction of Meloidogyne incognita and Rotylenchulus reniformis on Sweet Potato. AB - The influence of various factors on reproduction of concomitant Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) on sweet potato were studied in the greenhouse. Reproduction of Rr was reduced by Mi at all inoculum levels and experiment durations used, while Mi reproduction was not inhibited. Both species failed to affect each other when inoculated simultaneously onto root systems developed in separate pots from different nodes of the same plant. Reproduction of each species was not significantly greater when inoculation of the second species was delayed 1-2 weeks compared to simultaneous inoculation. After shoot excision, Rr increased in the soil but Mi decreased. Fibrous root weights of plants inoculated with Rr + Mi in some tests were higher than those inoculated with Mi alone, indicating an early suppression of Mi and/or root stintulation by Rr. Drought stress delayed Rr egg hatching and movement of larvae into the soil, but had little effect on Mi reproduction. PMID- 19295795 TI - Interactions between nematodes and other factors on plants. AB - The distinction between qualitative and quantitative interactions is stressed as it helps to elucidate differences between two current definitions of the term synergism and how multifactorial experiments should be designed, analysed statistically, and their results interpreted. Factorial design and analyses are strongly advocated so that interactions can be detected. It is suggested that interactions involving nematodes are common in nature and should be included, where appropriate, in hypotheses. Methods for testing the hypothesis that environment influences tolerance are suggested. PMID- 19295796 TI - Validation of a model for prediction of host damage by two nematode species. AB - PLANT ROOTS WERE MECHANICALLY INJURED OR SUBJECTED TO NEMATODE PARASITISM TO TEST THE MODEL OF HOST DAMAGE BY TWO NEMATODE SPECIES: y = m' + (l - m')c'z(P)z(P) for y 1.0, where m' = m + (m - m) (1 - y)/[(1 - y) + (l - y)] and c' = (z(-T) + z(-T))/2. Damage functions for greenhouse-grown radish plants (cv. Cherry Belle) mechanically injured with small or large steel needles were used to predict growth of plants injured by both needles. Growth predictions accounted for 94%, 87%, and 82% of mean treatment variation in plant height, stem weight, and root weight, respectively. Cowpea (cv. California Blackeye No. 5) damage functions, based on preplant population levels of Meloidogyne incognita and M. javanica, were used to predict seed yield of plants concomitantly infected with various levels of each species. Single species damage functions and population growth curves indicated significant host resistance to M. incognita and significantly lower virulence of that species compared to M. javanica. Model predictions accounted for 88% of mean seed yield variation in two-species treatments. In a separate experiment, mean top weights of 30-day-old cowpea plants, nniformly inoculated with 20,000 M. javanica eggs, increased with increasing levels of concomitantly inoculated M. incognita eggs. It is speculated that competitive interactions between M. incognita and M. javanica mitigated host damage by the more virulent species. PMID- 19295797 TI - Host-Parasite Relationships of Atalodera spp. (Heteroderidae). AB - Atalodera ucri, Wouts and Sher, 1971, and A. lonicerae, (Wouts, 1973) Luc et al., 1978, induce similar multinucleate syncytia in roots of golden bush and honeysuckle, respectively. The syncytium is initiated in the cortex; as it expands, it includes several partially delimited syncytial units and distorts vascular tissue. Outer walls of the syncytium are relatively smooth and thickest near the feeding site of the nematode; inner walls are interrupted by perforations which enlarge as syncytial units increase in size. The cytoplasm of the syncytium is granular and includes numerous plastids, mitochondria, vacuoles, Golgi, and a complex network of membranes. Nuclei are greatly enlarged and amoeboid in shape. Although more than one nucleus sometimes occur in a given syncytial unit, no mitotic activity was observed. Syncytia induced by species of Atalodera chiefly differ from those of Heterodera sensu lato by the absence of cell wall ingrowths; wall ingrowths increase solute transport and characterize transfer cells. In syncytia of Atalodera spp., a high incidence of pits and pit fields in walls adjacent to vasctdar elements suggests that in this case plasmodesmata provide the pathway for increased entry of sohttes. The formation of a syncytium by species of Atalodera and Heterodera sensu lato, but a single uninucleate giant cell by Sarisodera and Hylonema, indicates a pattern of host responses that may be useful, with other characters, for phylogenetic inference for Heteroderidae. PMID- 19295798 TI - Population Development of Pratylenchus hexincisus in Eight Corn Inbreds. AB - Of eight corn inbreds tested in the greenhouse and field, three (H60, H95, and H84) supported lower populations of Pratylenchus hexincisus than other inbreds included in this study. No apparent differences existed among inbreds in nematode invasion or development in the roots, or in population structure. Differences in population were therefore attributed to differential reproduction. PMID- 19295799 TI - Influence of Planting Date on Damage to Soybean Caused by Heterodera glycines. AB - Bragg soybeans were planted in nematicide-treated and nontreated plots on 15 May, 15 June, 1 July, and 15 July in 1980 and 1981 to determine the influence of planting date on damage caused by H. glycines. Although earlier studies showed the nematode was sensitive to high soil temperatures (> 34 C), late planting did not reduce damage caused by the nematode. Yields from plots treated with 1, 2 dibromo-3-chloropropane (57.5 kg a.i./ha) were 48, 118, 395, and 403% higher than yields from nontreated plots with planting dates of 15 May, 15 June, 1 July, and 15 July, respectively, when data were averaged over the 2 years. Increase in both seed size and number accounted for the yield increases in treated plots. Soil temperatures were highest during July in 1980, averaging 8.9 and 6.5 hours per day above 34 C at 10- and 20-cm depths, respectively. Larvae populations of H. glycines were reduced by the nematicide but not by late planting. These results indicate that damage caused by H. glycines may actually increase with later planting and that nematicides may be more beneficial when soybeans are planted late in a double-cropped production system. PMID- 19295800 TI - Host Response to Sarisodera hydrophila Wouts and Sher, 1971. AB - The histopathology of two populations of Sarisodera hydrophila Wouts and Sher, 1971 was examined on Salix lasiolepis Benth. (willow), Populus fremontii Wats. (cottonwood), and Lyonothamnus floribundus Gray (ironwood) using light microscopy as well as scanning and transmission electron microscopy. Sarisodera hydrophila induces formation of a single uninucleate hypertrophied cell (giant cell) which varies only slightly among the three hosts. The giant cell is enclosed by the root stele and contacts phloem, vascular cambium, and xylem. The single hypertrophied nucleus of the giant cell is ameboid or lobulate in shape, generally with a single nucleolus. The cell is characterized by a wall which is separated into two distinct regions about 2 mum and 13 mum thick; the thicker region occurs adjacent to the nematode head. Cell wall ingrowths, such as those associated with host responses to certain other plant-parasitic nematodes, were not observed in giant cells induced by S. hydrophila. However, a high frequency of pit fields with plasmodesmata occurred in the thinner portion of the cell wall which is adjacent to vascular elements. Roots of the three hosts simultaneously infected with S. hydrophila and Meloidogyne sp. resulted in adjacent responses characteristic of each nematode, supporting the view that the specific type of host response is a function of the nematode rather than the host. The varying expressions of host responses among Heteroderoidea may be useful in testing congruency with existing interpretations of phylogeny. PMID- 19295801 TI - Effect of Sesbania rostrata on Hirschmanniella oryzae in Flooded Rice. AB - Microplot experiments on flooded soil infested with Hirschmanniella oryzae were conducted to investigate the influence of the legum Sesbania rostrata as a rotation crop with rice, Oryza sativa L. cv. Moroberekan. To avoid a green manure effect from S. rostrata, all aerial parts were removed at harvest. The dry weight of paddy, culms and leaves, and number of culms of rice following Sesbania were 214%, 158%, and 121% greater, respectively, than those following rice. Ripening of the paddy occurred earlier if rice followed Sesbania. The beneficial effect of Sesbania may have been due to the trap-crop action of Sesbania against H. oryzae. PMID- 19295802 TI - Development of Heterodera glycines Ichinohe on Soybean, Glycine max (L.) Merr., under Gnotobiotic Conditions. AB - The life cycle of the soybean cyst nematode, Race 3 (SCN 3), Heterodera glycines Inchinohe was determined from observations of the developmental stages on soybean Glycine max cv. Kent root explants under gnotobiotic conditions at 25 C. Approximately 51% of the second-stage larvae penetrated the root l day after inoculation (DAI). Third-stage larvae appeared 4 DAI, became sexually differentiated 5 DAI, and protruded from the root tissues 6 DAI. Fourth-stage males and females were observed 7 DAI. Ensheathcd adult males were observed at 9 DAI and exsheathed to free adults at 11 DAI. The fourth-stage female became an adult at l0 DAI, Males entwined arotmd the gelatinons sac of the female at 12 DAI and were assumed to be mating. Some males actually penetrated and were enveloped by the gelatinous sac. The female-to-male sex ratio ranged from 2.3 to 9.5:1. First- and second-stage larvae were observed in the egg 17 and 19 DAI, respectively. The life cycle of the SCN 3 was completed 21 DAI upon hatching of the eggs and emergence of second-stage larvae. The average number of eggs in the cyst body and gelatinous sac, was 210 and 187, respectively. Key words: reproduction, soybean cyst nematode, scanning electron microscopy. PMID- 19295803 TI - Susceptibility of plant selections to Heterodera schachtii and a race of H. trifolii parasitic on Sugarbeet in The Netherlands. AB - Similar host ranges were found for Heterodera schachtii and a race of H. trifolii parasitic on sugarbeet in The Netherlands. Twenty-nine of 41 plant accessions evaluated were susceptible to H. trifolii. Five breeding lines of the interspecific hybrid Beta vulgaris-B. procumbens which are resistant to H. schachtii were highly susceptible to H. trifolii. An accession of B. maritima with partial resistance to H. schachtii was resistant to H. trifolii. PMID- 19295804 TI - Development of the False Root-knot Nematode, Nacobbus aberrans, on Sugarbeet. AB - The duration of the embryogenic development of Nacobbus aberrans (= N. batatiformis) took 9-10 days at 25 C and 51 days at 15 C. The J molted in the egg; hence the Je emerged from the egg. The effect of distilled water attd root leachates of kochia and sugarbeet was investigated at 5, 10, 15, 20, and 25 C. Root leachates did not significantly affect the percent of cumulative hatch of eggs, but temperature did significantly affect emergence of juveniles (p = 0.05). Less than 1, 5, and 20% of eggs hatched at 5, 10, and 15 C, respectively. The percent of cumulative hatch at 20 C was four times greater than at 15 C, while the highest percentage of juveniles emerged at 25 C. The duration of postembryogenic development from J inoculation until the appearance of mature females with egg masses took 38 days, and the life cycle from egg to egg was completed in 48 days at 25 C. All immature stages, young females and males were migratory endoparasites. Young females were able to leave the root swellings, where they developed from juvenile stages, and re-enter the root, where they formed a true gall and became sedentary. Thirty days after inoculation with J nematodes, specimens were detected in root tissues at 10, 15, 20, 25, and 30 C, hut not at 5 C. Five days after inoculation at 23 C ( +/- 2 C), juveniles had penetrated the roots and caused slight swellings of the tip and axis of sugarbeet feeder roots. Large cavities extended from the cortical parenchyma to the periphery of the stelar area, and 50 % of the central cylinder was destroyed 25 days after inoculation at 23 C. No syncytia formation were detected in the sugarbeet root swellings infected with juveniles. Syncytia were associated only with adult females; hyperplasia, abnormal proliferation of lateral roots, and asymmetry of root structure were additional anatomical changes induced by adult females. Only very smooth annules but no cuticular ornamentations were noted by SEM on the perineal area of adult females. PMID- 19295805 TI - Development of Thelastoma bulhoesi (Oxyurata: Thelastomatida) and the Effect of Thiabendazole on the Unembryonated Egg. AB - The embryological and postembryological development of Thelastoma bulhoesi was determined. Initial cleavage was into unequal cells and occurred within 1-2 hours at 25 C. Cell division was holoblastic but no true morula is formed. Gastrulation occurred at approximately 48 hours by epibolic synectic mechanisms. First-stage larvae were fully developed at 96 hours. The molt to second-stage larvae was initiated in the egg and was completed at hatching. Second-stage larvae were first observed in the host at 11 hours postinfection, third-stage larvae at 18 hours, and fourth-stage larvae at 192 hours. Adult female worms were observed at 32 days. Thiabendazole, in even the lowest concentrations, inhibited the developmem of unembryonated ova. PMID- 19295806 TI - Parasitic Variability of Meloidogyne incognita Populations on Susceptible and Resistant Cotton. AB - Root gall induction and egg production by the four recognized host races and two cytological races of Meloidogyne incognita were compared on cotton Gossypium hirsutum cvs. Deltapine 16 (root-knot susceptible) and Auburn 634 (highly resistant). The 12 nematode populations included in the study were from various parts of the world. No population increases occurred on the highly resistant cultivar. After 45 days, populations of host races 1 and 2 induced slight root galling on both cuhivars with only limited reproduction. Host race 4 populations induced moderate root galling with higher reproduction on Deltapine 16 than that of race 1 or race 2 populations. Host race 3 populations induced severe root galling with population density increases of 7-30-fold. In a complementary study, 24 cotton cultivars or breeding lines were compared for suitability as hosts for a typical population of M. incognita race 3. The poorest hosts, 'Aubnru 623,' 'Auburn 634,' and 'McNair 220,' yielded fewer eggs after 45 days than were added initially. The best hosts - 'M-8.' 'DES 24-8,' 'McNair 235,' and 'Coker 20l' - yielded > 5 times as many eggs as were added initially. PMID- 19295807 TI - Effect of Soil Texture on the Distribution and Infectivity of Neoaplectana carpocapsae (Nematoda: Steinernematidae). AB - The vertical migration of N. carpocapsae infective juveniles applied to the soil surface or introduced 14 cm below the soil surface was studied in four different soil types (pure silica sand, coarse sandy loam, silty clay loam, and clay). The percentage of juveniles able to migrate and infect wax moth pupae placed in the soil decreased as the percentage of clay and silt increased. Most nematodes placed on the soil surface remained within 2 cm of the surface, but some penetrated to a depth of 10 cm in pure silica sand and coarse sandy loam to infect pupae. Some pupae at the same depth were also infected with nematodes in silty clay loam soil. In pure silica sand and coarse sandy loam, nematodes introduced 14 cm below the soil surface were able to infect wax moth pupae located between 4 and 24 cm. Movement was least in clay soil and limited in silty clay loam. Nematodes showed a tendency to disperse upwards from the point of application. In all cases the number of migrating nematodes was greatest when wax moth pupae were present. PMID- 19295808 TI - Nematode parasites and associates of Dendroctonus spp. and Trypodendron lineatum (Coleoptera: Scolytidae), with a description of Bursaphelenchus varicauda n.sp. AB - Nematode parasites and associates of four bark beetle species in British Columbia were surveyed. Bursaphelenchus varicauda n.sp., Ektaphelenchus macrostylus, Panagrolaimus dentatus, and Cryptaphelenchus latus were found associated with Dendroctonus pseudotsugae. Parasitorhabditis obtusa was found in the gut and Contortylenchus reversus in the hemocoel of both D. pseudotsugae and D. rufipennis. The latter also had hemocoel infections of Sphaerulariopsis dendroetoni, which were not found concomitant with C. reversus infections. Contortylenchus brevicomi occurred in the hemocoel of D. brevicomis. The first report of a tylenchid larva parasitizing Trypodendron lineatum in North America is presented. Bursaphelenchus varicauda n.sp. was obtained from the gallery frass of D. pseudotsugae. It resembles B. corneolus and B. bestiolus but differs from the former species in female tail shape, the position of the excretory pore, spicule shape, and position of the male caudal papillae, and from the latter species in spicule shape and in the length of the esophagus and postuterine sac. PMID- 19295810 TI - Hypoaspis nr. aculeifer: a Mite Predacious on Root-knot and Cyst Nematodes. PMID- 19295809 TI - Plant-parasitic nematodes of South viet nam. AB - Between 1974 and 1978, 2,842 identifications of plant-parasitic nematodes were made from more than 1,700 soil and plant samples collected in eight provinces of South Viet Nam. Species in nine genera-Helicotylenchus, Criconemoides, Meloidogyne, Pratylenchus, Tylenchorhynchus, Hoplolaimus, Hirschmanniella, Xiphinema, and Rotylenchulus-comprised 96.1% of the identifications; the remaining 3.9% were species of 11 genera. Fourteen genera were associated with rice which was grown on about 2,500,000 ha in 1970. Of these, Ditylenchus, Hirschmanniella, and Meloidogyne were most important. Ditylenchus angustus caused severe damage to about 50,000 ha of flooded rice in the Mekong Delta in 1976. Hirschmanniella spp. were found in all samples examined from flooded rice fields. Meloidogyne spp. were common in rice seedbeds, upland rice, and rice not kept flooded continuously. Meloidogyne and Pratylenchus spp. were found in roots of 22 of the 32 crop plants sampled. Little or no attempt was made in South Viet Nam to control nematodes. PMID- 19295811 TI - Wall Structure of the Mature Female and Cystoid Body of Meloidoderita kirjanovae (Nematoda: Criconematoidea). PMID- 19295812 TI - Effect of Soil Texture on the Distribution and Infectivity of Neoaplectana glaseri (Nematoda: Steinernematidae). AB - The vertical migration of infective juveniles of Neoaplectana glaseri applied to the soil surface or introduced 16 cm below the soil surface was studied in pure silica sand, coarse sandy loam, silty clay loam, and clay. The number of juveniles that migrated and infected wax moth pupae placed in the soil decreased as the proportion of clay and silt increased. The majority of nematodes moved downwards 2-6 cm from the surface, but some penetrated to a depth of 14 cm in pure silica sand and coarse sandy loam. In pure silica sand and coarse sandy loam, nematodes introduced 16 cm below the soil surface were able to infect wax moth pupae located at depths of 0-4 cm and 28-32 cm. Nematodes showed a greater tendency to disperse downwards from the point of application. Movement of the nematode was least in clay soil and limited in silty clay loam soil. The number of migrating nematodes was greatest when wax moth pupae were present. PMID- 19295813 TI - Studies on Anhydrobiosis of Pratylenchus thornei. AB - Large populations of Pratylenchus thornei, a winter pest of cereals, legumes, and potatoes in the northern Negev region of Israel, survive 7-8 months of summer drought and return to full activity at the beginning of the rainy season. To demonstrate that it survives the summer in an anhydrobiotic state, all developmental stages of P. thornei were exposed to gradually reduced relative humidity (RH) using glycerin water solutions. At 97.7% RH the nematodes were coiled and able to survive exposure to 0% RH. About 40% of artificially desiccated nematodes could be reactivated by gradually increasing the humidity to the final water environment. Desiccated nematodes could withstand temperatures up to 40 C. Reactivated individuals showed intestines apparently devoid of reserve materials. Only 3% survived three cycles of desiccation and reactivation. P. thornei reactivated after anhydrobiosis multiplied twice as much within Vicia sativa roots as did fresh nematodes. PMID- 19295814 TI - Biochemical Identification of the Two Races of Radopholus similis by Starch Gel Electrophoresis. AB - Analysis of genetic variation between the banana and the citrus races of Radopholus similis by starch gel eleclrophoresis demonstrated that 7 of 16 enzyme encoding loci could be used for their diagnostic separation. The two races are closely related arid share approximately 75% of the enzymes evaluated. The level of dissimilarities o1 inherited bands indicates that no gene flow occurs between the races. Aldolase, alpha + beta esterase, glucose-6-phosphate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase, malate dehydrogenase, and phosphoglucose isomerase are diagnostic markers of the races. PMID- 19295815 TI - Biochemical Identification of the Two Races of Radopholus similis by Polyacrylamide Gel Electrophoresis. AB - Analysis of proteins of the banana and citrus race of Radopholus similis was carried out by several different types of polyacrylamide gel electrophoresis. These included standard slab gel, SDS slab gel, gradient slab gel, and two ditnensional slab gel electrophoresis. A major band difference was detected between the two races by slab gel electrophoresis. However, several other poorly resolved but consistent hands of high molecular weight proteins near the gel origin also were considered as diagnostic. Resolution of protein bands was greatly improved by SDS and gradient slab gel electrophoresis, but no differences could be detected among the proteins resolved between the two rares with these techniques. Two-dimensional gels revealed a large number of proteins, but background staining obscured them hindering interpretation. When nematode races were reared on three different host plants, no differences in protein patterns were detected between them, indicating host preferences does not play a role in determining the types proteins occurring in these nematodes. PMID- 19295816 TI - Meloidogyne aquatilis n. sp (Nematoda:Meloidogynidae) from Spartina pectinata with a Key to the Canadian Species of Meloidogyne. AB - A new root-knot nematode, Meloidogyne aquatilis n. sp., attacking the roots of Spartina pectinata Link growing in the Ottawa River is described and illustrated. Meloidogyne aquatilis is distinguished from M. graminis by the light brown body color and by the absence of perineal lateral fields in the female. The male differs by the shorter stylet and by the hemizonid being separated by 7-9 annules from the excretory pore. The second-stage juveniles are also recognized by the 7 9-annule gap between the hemizonid and excretory pore and by the shorter tail with a disc-like subterminal tail structure, lower b ratio value, and inflated rectum. A key to the root-knot nematode species of Canada based on females, males, and juveniles is provided. The type host of Dolichodera fluvialis Mulvey and Ebsary, 1980, Spartina pectinata Link, is reported for the first time. PMID- 19295817 TI - Description of Pararotylenchus belli n.sp. (Nematoda:Hoplolaimidae). AB - A new species of Pararotylenchus Baldwin &Bell, 1981 from the Pacific coastal area of California is described. Numerous females and juveniles were recovered, but no males were found. Pararotylenchus belli n.sp. is most similar to P. sphaerocephalus, but females of P. belli are shorter and have a shorter stylet and a hemispherical tail. Specimens from a cereal field in South Dakota were identified as P. colocaudatus. PMID- 19295818 TI - Description of Thecavermiculatus andinus n.sp. (Meloidoderidae), a Round Cystoid Nematode from the Andes Mountains of Peru. AB - Thecavermiculatus andinus n.sp. is described and illustrated from Oxalis tuberosa originally collected in the vicinity of Lake Titicaca high in the Andes mountains of southern Peru. This new species differs markedly front the other two species in the genus, especially in having a much greater female vulval-anal distance and annules with lined punctation on most of the female body with a lacelike pattern restricted to the posterior portion, particularly at the vulva and anus which do not protrude. Females are essentially spherical with protruding neck, white to yellowish in color, and can easily be mistaken for potato cyst nematodes. Among the dozen or more known weed and crop host plants are potato and eggplant. In order to accommodate this new species, the genus Thecavermieulatus is emended. A key to the species of this genus is presented. PMID- 19295819 TI - Pungentus porosus n.sp. (Nematoda:Dorylaimoidea) from Canada with a Key to the Species of Pungentus. AB - Pungentus porosus n. sp. is described and illustrated. The differential characters are a short odontostyle of 6-8 mum or slightly less than one head width, vulva at 44-50%, goblet-shaped amphids, presence of a prevulval uterine sac, and a hemispherical tail with a variable number of pores. Pungenlus sublatum, P. vesiculosus, P. orthocephalus, P. granosus, and P. parvus sensu Eroshenko are considered to be incertae sedis. A key is provided to the 21 valid species of Pungentus. The following species are new Canadian records: P. angulosus, P. clavatus, P. engradinensis, P. marietani, P. microdentatus, P. monohystera, P. obscurus, P. parvus, P. pungens and P. silvestris. PMID- 19295820 TI - Morphological Characteristics of Hoplolaimus indicus Sher, 1963 in Canada, a Parasite of Wild Rice. AB - Hoplolaimus indicus Sher, 1963 was found for the first time in North America as a parasite of wild rice. Supplemental descriptive data, supported by illustrations, are given which extend the known range of variability in this species. The body, stylet, and tail lengths were often greater than those in published reports from India, the esophageal gland overlap was consistently shorter than body width in most specimens, and the excretory pore and vulva were more posterior. Longitudinal striae of the basal head annule ranged from 6 to 20, number of tail atmules from 8 to 22, and the excretory pore extended at most 27 mum anterior or 22 mum posterior of the esophagointestinal valve. A taxonomic key is provided to facilitate the separation of the 13 species of Hoplolaimus having five or six esophageal gland nuclei. PMID- 19295821 TI - Fine structure of body wall cuticle of females of Meloidodera charis, Atalodera lonicerae, and Sarisodera hydrophila (Heteroderidae). AB - The body wall cuticle of adult females of Meloidodera charis, Atolodera lonicerae, and Sarisodera hydrophila is examined by transmission electron and light microscopy for comparison with Heterodera schachtii and previous observations of additional species of Heterodera, Globodera, and Punctodera. The cuticle of M. charis is least complex, consisting of layers A, B, C (with A outermost), and varies in overall thickness from 3 to 8 mum. As in other species, the cuticle is thickest in mature specimens. The cuticle of A. lonicerae is 6-9 mum thick; unlike M. charis it has an innermost layer, D, in addition to A, B, and C. The cuticle of S. hydrophila varies from 14 to 30 mum thick and includes a D layer similar to A. lonicerae; layer C is subdivided into additional zones relative to other heteroderids, and the external portion of the cuticle is infused with an electron-dense material. The presence of a D layer in A. lonicerae and S. hydrophila is a character state which is shared with Globodera spp. and Punctodera sp. The electron-dense material in the outer layers of S. hydrophila also occurs in Globodera spp. and Punctodera sp. On the other hand, H. schachtii resembles other Heterodera spp. as well as M. charis by the absence of a D layer and lack of electron-dense material in the outer layers. The pattern of occurrence of shared character states, including those of the cuticle, may be useful for phylogenetic analysis of Heteroderidae. PMID- 19295822 TI - Meloidogyne enterolobii n. sp. (Meloidogynidae), a Root-knot Nematode Parasitizing Pacara Earpod Tree in China. AB - Meloidogyne enterolobii n. sp. is described and illustrated from roots of pacara earpod tree, Enterolobium contortisiliquum (Vell.) Morong, on Hainan Island in China. The perineal pattern of the female is usually oval shaped, the striae are fine to coarse, the dorsal arch is moderately high to high and usually rounded, and the phasmids are large. The stylet knobs in females are divided longitudinally by a groove so that each knob appears as two. The mean distance of the excretory pore to the anterior end in the female is 62.9 mum. Males have a large, rounded labial disc that fuses with the medial lips to form a dorso ventrally elongate head cap. The labial disc is slightly elevated, and the medial lips are crescent shaped. The second-stage juvenile mean body length is 436.6 mum. The lateral lips are large and triangular in face view. The tail is 56.4 mum long and narrow with a broad, bluntly rounded tip. M. enterolobii n. sp reproduces well on E. contortisiliquum and causes severe damage. Other good hosts include cotton, resistant tobacco 'NC 95,' pepper, watermelon, and tomato. PMID- 19295823 TI - Histopathology of Selected cultivars of Tobacco infected with Meloidogyne species. AB - Rates of nematode penetration and the histopathology of root infections in fluecured tobacco cultivars 'McNair-944,' 'Speight G-28,' and 'NC-89' with either Meloidogyne arenaria, M. incognita, M. hapla, or M. javanica were investigated. Penetration of root tips by juveniles of all species into the M. incognita resistant NC-89 and G-28 was much less than that on the susceptible McNair-944. Few juveniles of M. incognita were detected in resistant cultivars 7 and 14 days after inoculation. Infection sites exhibited some cavities and extensive necrotic tissue at 14 days; less necrotic tissue and no intact nematodes were observed 35 days after inoculation. Although some females of M. arenaria reached maturity and produced eggs, considerable necrosis was induced in the resistant cultivars. Meloidogyne hapla and M. javanica developed on all cultivars, but there was necrotic tissue at some infection sites in the resistant cultivars. The occurrence of single multistructured nuclei in the syncytia of most M. hapla infections differed from the numerous small nuclei found in syncytia caused by the other three species. PMID- 19295824 TI - Influence of Maize Rotations on the Yield of Soybean Grown in Meloidogyne incognita Infested Soil. AB - A replicated field study was conducted from 1972 to 1980 involving soybeans grown in 2-, 3-, and 4-year rotations with maize in soil infested with Meloidogyne incognita. Monocultured soybeans were maintained as controls. Cropping regimes involved root-knot nematode susceptible and resistant soybean cultivars and soybeans treated and not treated with nematicides. Yields of susceptible cultivars declined with reduced length of rotation. Nematicide treatment significantly increased yields of susceptible cultivars when monocultured, but bad little influence on yield when susceptible cultivars were grown in rotation. Yields of monocultured resistant cultivars were significantly lower than yields of resistant cultivars grown in rotation. However, yields of resistant cultivars grown in rotation were not influenced by the length of the rotation. Nematicide treatment significantly increased yields of monocultured resistant cultivars over the latter years of the study. PMID- 19295825 TI - Resistance of Some Vitis Rootstocks to Xiphinema index. AB - Thirty-eight grapevine (Vitis spp.) rootstocks were screened in pots for resistance to the dagger nematode, Xiphinema index, from 1979 to 1981. Resistance ratings were based on visible root symptoms and on changes in the nematode populations over 16 months. Nineteen of the 23 Californian hybrid rootstocks tested were resistant, as were 'Harmony',' 'Freedom,' 'Schwarzmann,' and '3309.' Two hybrids of V. rufotomentosa, '171-52' and '176-9,' were possibly immune to X. index. The rootstocks 'ARG 1,' ' 110 R,' '1202,' and '1616,' which are used commercially for phylloxera resistance were susceptible. PMID- 19295826 TI - Interaction of Vesicular-arbuscular Mycorrhizal Fungi and Phosphorus with Meloidogyne incognita on Tomato. AB - The influence of two vesicular-arbuscular mycorrhizal fungi and phosphorus (P) nutrition on penetration, development, and reproduction by Meloidogyne incognita on Walter tomato was studied in the greenhouse. Inoculation with either Gigaspora margarita or Glomus mosseae 2 wk prior to nematode inoculation did not alter infection by M. incognita compared with nonmycorrhizal plants, regardless of soil P level (either 3 mug [low P] or 30 mug [high P] available P/g soil). At a given soil P level, nematode penetration and reproduction did not differ in mycorrhizal and nonmycorrhizal plants. However, plants grown in high P soil had greater root weights, increased nematode penetration and egg production per plant, and decreased colonization by mycorrhizal fungi, compared with plants grown in low P soil. The number of eggs per female nematode on mycorrhizal and nonmycorrhizal plants was not influenced by P treatment. Tomato plants with split root systems grown in double-compartment containers which had either low P soil in both sides or high P in one side and low P in the other, were inoculated at transplanting with G. margarita and 2 wk later one-half of the split root system of each plant was inoculated with M. incognita larvae. Although the mycoorhizal fungus increased the inorganic P content of the root to a level comparable to that in plants grown in high P soil, nematode penetration and reproduction were not altered. In a third series of experiments, the rate of nematode development was not influenced by either the presence of G. margarita or high soil P, compared with control plants grown in low P soil. These data indicate that supplemental P (30 mu/g soil) alters root-knot nematode infection of tomato more than G. mosseae and G. margarita. PMID- 19295827 TI - Dispersion and Distribution of Pratylenchus scribneri and Hoplolaimus galeatus in Soybean Fields. AB - Examination of dispersional characteristics of Pratylenchus scribneri and Hoplolaimus galeatus indicated that there were patches within soybean fields in which both survival and reproduction wexe enhanced in spite of apparent homogeneity of soil type and topography. Treatment with carbofuran reduced the patchiness (or increased the dispersion) for H. galeatus while it had the opposite effect for P. scribneri. P. scribneri was less highly dispersed in conventional tillage plots than in the zero tillage plots. Populations from quadrats contained entirely within the patches could be described by the normal distribution (in the case of P. scribneri) or by the Poisson distribution (in the case of H. galeatus), while populations from quadrats contained entirely outside the patches could be described by the Poisson distribution for both nematodes. None of the distributions tested (Poisson, normal, negative binomial, Neyman's) gave an adequate fit when populations from both inside and outside the patches were considered together. In all instances, log and ln transformations reduced the goodness of fit of the data to all of the distributions tested. Even with logarithmic transformations, parametric statistics were not appropriate for analysis of data in most instances. PMID- 19295828 TI - The Interrelationship of Heterodera schachtii and Ditylenchus dipsaci on Sugarbeet. AB - Heterodera schachtii significantly (P = 0.05) reduced sugarbeet root growth below that of uninoculated controls at 20, 24, and 28 C, and Ditylenchus dipsaci significantly (P = 0.05) reduced root growth below that of uninoculated controls at 16, 20, 24, and 28 C. A combination of H. schachtii and D. dipsaci significantly (P = 0.05) reduced root growth below that of single inoculations of H. schachtii at all temperatures and D. dipsaci at 20, 24, and 28 C. Single inoculations of H. schachtii and D. dipsaci significantly (P = 0.05) reduced top growth of sugarbeet below that of uninoculated controls at 20, 24, and 28 C, and 16, 20, 24, and 28 C, respectively. A combination of the two nematodes significantly (P = 0.05) reduced top growth below that of single inoculations of H. schachtii at all temperatures. However, a combination of the two nematodes failed to significantly (P = 0.05) reduce top growth below that of single inoculations of D. dipsaci at any temperature. Inoculations of either H. schachtii or D. dipsaci did not affect penetration of the other nematode, and D. dipsaci did not affect development and reproduction of H. schachtii. D. dipsaci did not reproduce on sugarbeet. PMID- 19295829 TI - Population Dynamics of Heterodera glycines and Soybean Response in Soils Treated with Selected Nematicides and Herbicides. AB - Two field experiments were conducted in two locations to determine the effects of the nematicides aldicarb, phenamiphos, and ethoprop and/or the herbicides alachlor, linuron, or metribuzin on the population dynamics of Heterodera glycines and soybean growth and yield. Population densities of H. glycines were greater, at some time during the growing season, in several treatments with alachlor alone and in combination with nematicides. Numbers of H. glycines at harvest were greater in plots treated with aldicarb than in those treated with ethoprop or phenamiphos. The numbers in aldicarb treated plots were generally reduced when plots also received a herbicide. Soybean yields were negatively correlated with numbers of H. glycines eggs and juveniles in early to mid season but positively correlated with late season population densities. PMID- 19295830 TI - On the methodology of nematode extraction from field samples: baermann funnel modifications. AB - Routine quantitative nematode extraction for pest management purposes remains a problem. There is need for more knowledge of the parameters limiting efficiency of the various available methods. Sedimentation rates for several species of nematodes have been confirmed as slow and highly variable and therefore not suitable for quantitative separation of nematodes. Funnel losses with clean and unpitted glassware, whether closed or open stemmed, with or without misting, are negligible so long as misting periods are neither inadequate nor excessive; i.e., approximately a 1.5-min water spray period in a 10-min cycle. Tissue paper used to retain soil, sievings, or other substrate in the funnel extraction can greatly inhibit the passage of nematodes depending upon the tissue properties and the nematode species. PMID- 19295831 TI - On the methodology of nematode extraction from field samples: density flotation techniques. AB - Density flotation has been frequently used for the extraction of nematodes from field samples. Density flotation curves for four nematode species and five solutes have been prepared. The curves confirm that flotation was governed by several factors: solute density, solute osmotic activity, and physiological properties of the nematode species. Nematode viability and function can be adversely affected by improper selection of solute for density extraction of nematodes; nevertheless, some nematode species can be enriched from mixtures by density and solute selection. PMID- 19295832 TI - On the methodology of nematode extraction from field samples: comparison of methods for soil extraction. AB - The commonly used nematode extraction methods were compared using three soil types and four nematode species. The comparison was repeated in three trials by the same operator to estimate operator reproducibility. Extraction efficiency was dependent upon method, soil type, and nematode species, and reproducibility was not particularly satisfactory for routine analyses. Extraction by any method tested was less than 50% efficient. Quantitative nematode extraction methodology needs serious attention. PMID- 19295833 TI - Influence of metalaxyl on three nematodes of citrus. AB - Metalaxyl significantly reduced population of Pratylenchus coffeae, Radopholus similis, and Tylenchulus semipenetrans in roots of Citrus limon (rough lemon) under greenhouse conditions. Postinoculation treatment of rough lemon seedlings was not as effective i n reducing nematode populations as was treatment before inoculation. Fewer nematodes infected metalaxyl-treated roots than nontreated roots. However, incubation of nematodes in metalaxyl did not inhibit nematode motility or their ability to locate and infect roots. Cellular responses to nematode injection differed between treated and nontreated tissues. Metalaxyl appeared to confer nematode contraol by modifying citrus roots such that a normally susceptible rootstock became tolerant. PMID- 19295834 TI - Rhizonema sequoiae n.gen. n.sp. from Coast Redwood Sequoia sempervirens (D.Don) Endl. AB - Rhizonema sequoiae n.gen, u. sp. is described from the roots of Coast Redwood, Sequoia sempervirens (D. Don) Endl., growing near Lake Lagunitas, Marin County, California. Rhizonema females are annulated over their entire bodies, are wholly embedded in host tissue, and secrete an abundant amount of gel material. Mature females do not form a cyst. The vulva is located on a large posterior terminal cone, and the anus is on the dorsal vulval lip. Esophageal glands of the second stage larvae fill more than half of the body cavity. Tails of the vermiform males are blunt, and a cloacal tubus is present. PMID- 19295835 TI - Effects of Selected Nematicides on Hatching of Heterodera schachtii. AB - Aldicarb, carbofuran, fensulfothion, and phenamiphos were tested in concentrations of 1-100 mug/ml for their effects on hatching of Heterodera schachtii. Exposure of cysts to 1 mug aldicarb or carbofuran/ml stimulated hatch whereas phenamiphos and, to a lesser degree, fensulfothion inhibited hatch. Addition of aldicarb to sugarbeet root diffusate or 4 mM zinc chloride suppressed activities of these hatching agents. Transfer of cysts previously treated with aldicarb or carbofuran to zinc chloride or water rapidly initiated hatch which finally exceeded the hatch from cysts not treated with the nematicides. PMID- 19295836 TI - Control of Globodera rostochiensis in Relation to Method of Applying Nematodes. AB - Soaking potato tuber pieces for 15 min in 8,000 mug/ml of oxamyl just before planting reduced the number of Globodera rostochiensis cysts that developed on potato roots, but this treatment was phytotoxic. Five foliar applications of 1.12 kg a.i./ha of oxamyl or carbofuran at 10-day intervals beginning when 90% of the plants had emerged suppressed increase in G. rostochiensis densities. Similar foliar applications of phenamiphos were ineffective in controlling G. rostochiensis. Soil applications (in the row at planting) of aldicarb, carbofuran, phenamiphos, ethoprop, and oxamyl at 5.6 kg a.i./ha reduced the numbers of white females that developed on potato roots, but only those treatments involving aldicarb and oxamyl suppressed G. rostochiensis population increase. Combined soil and foliar treatments did not provide any advantage over soil treatment alone, as soil applications of 5.6 kg a.i./ha alone were equal to, or better than, combined soil (3.4 kg a.i./ha) and foliar (2.2 kg a.i./ha) applications in controlling G. rostochiensis. PMID- 19295837 TI - Effects of Dosage Sequence on the Efficacy of Nonfumigant Nematicides, Plantain Yields, and Nematode Seasonal Fluctuations as influenced by Rainfall. AB - Four nonfumigant nematicides applied three times during the wet season were used to study dosage sequence and nematicide effectiveness. Control of Helicotylenchus multicinctus (Cobb) Thorne and Meloidogyne javanica (Treub) Chitwood increased plantain (Musa AAB) yields. The nematicide (aldicarb, carbofuran, oxamyl, and miral) performance and yield response varied with dosage sequences. Applications of 2, 3, and 2 g ai/tree in March, July, and October (sequence I), respectively, gave greater control of M. javanica than did applications of 3, 2, and 2 g ai/tree in March, June, and September (sequence II), respectively. However, the high initial dose sequence was effective against H. multicinctus. Persistence of the different nematicides differed over the 14-month experimental period. Miral, aldicarb, and carbofuran were the most effective treatments against either species by the end of the wet and dry seasons. Dry season residual nematode populations were significantly lower in nematicide treated than in control plots. Yield increases over controls were 96.9, 90.1, 78.4, and 70.1% for carbofuran applied by sequence II, aldicarb by II and I, and oxantyl by II, respectively. Nematode populations directly fluctuated with rainfall and dropped to low (H. multicinctus) or to undetectable (M. javanica juveniles) levels during the dry season. Of the two nematodes studied, the more serious pest to plantain was H. multicinctus; it was tolerant to drought and survived the dry season in untreated soils. PMID- 19295838 TI - Use of Avermectins for the Control of Meloidogyne incognita on tomatoes. AB - The efficacy of avermectins B and B for control of Meloidogyne incognita on tomato was studied in pots and field plots for two seasons. Avermectins were applied as granules and liquid in furrows or by low pressure drip irrigation systems, at rates ranging from 0.093 to 0.34 kg a.i./ha, as single or multiple applications. Levels of control comparable to those obtained by oxamyl and aldicarb at 3.36 kg a.i./ha were achieved by the avermectin with only 1/10 the volume of chemicals applied to the environment. Avermectin protection of the roots remained constant throughout the first 5 weeks giving slightly longer protection than oxamyl or aldicarb. PMID- 19295839 TI - Effect of Aldicarb, Ethoprop, and Carbofuran on Control of Coffee Root-knot Nematode, Meloidogyne exigua. AB - Egg hatch of Meloidogyne exigua was significantly inhibited in 14 days pretreatment with aldicarb, ethoprop, or carbofnran at concentrations higher than 0.1 mug/ml; these eggs were found to delay hatch in 19 days posttreatment in ethoprop. Aldicarb and carbofuran solutions at concentrations greater than 0.1 mug/ml significantly decreased the motility and the life span of the second-stage juveniles; aldicarb was more toxic than carbofuran to the nematode. In a field test, aldicarb (Temik 10G), ethoprop (Mocap 10G), and carbofuran (Furadan 5G and Furadan Liquid 350F) significantly decreased M. exigua populations. PMID- 19295840 TI - Parasitic Habits of Trophotylenchulus floridensis (Tylenchulidae) and its Taxonomic Relationship to Tylenchulus semipenetrans and Allied Species. AB - Parasitism by Trophotylenchulus floridensis Raski, 1957 was studied on roots of sand pine (Pinus clausa [Chapm.] Vasey). Different life stages of the nematode were observed to be covered by dark, spherical, brittle, capsule-like structures which protruded from the root surface. The mature capsule enveloped a single sedentary female with a gelatinous matrix containing an average of 76 (44-117) eggs. The capsule was composed of a single layer of cells which appeared to be of plant origin. The anterior end of the nematode was embedded superficially in the root tissue where it created a feeding site comprised of a small number of discrete stelar parenchyma cells with dense cytoplasm and enlarged nuclei and nucleoli. The nematode also infected slash pine, Pinus elliottii, loblolly pine, P. taeda, red oak, Quercus falcata, post oak, Q. stellada, and sweet gum, Liquidambar styraciflua, in four different locations in central Florida. The taxonomic relationship between Tr. floridensis and Tylenchulus spp. is discussed. Based on differences in the tail and lip regions, position of the excretory pore, type of obesity and especially in the mode of plant parasitism, the genus Trophotylenchulus Raski, 1957 is upheld, and the transfer of Tylenchulus clavicaudatus Colbran, 1966, Ty. rnangenoti Luc, 1957, and Ty. obscurus Colbran, 1961 to Trophotylenchulus is proposed. PMID- 19295841 TI - Studies on Lasioseius scapulatus, a Mesostigmatid mite predaceous on nematodes. AB - The life history and feeding habits of Lasioseius scapulatus, an ascid predator and potential biocontrol agent of nematodes, was examined. Reproduction was asexual, and the life cycle was 8-10 days at room temperature. Life history consisted of the egg, protonymph, deutonymph, and adult. Both nymphal stages and the adult captured and consumed nematodes. Two fungal genera and eight genera of nematodes were suitable food sources. Second-stage root-knot nematode juveniles were eaten, but eggs and adult females were not. The mite fed voraciously on nematodes and drastically reduced Aphelenchus avenae populations in vitro. It is suggested that mites are of considerable importance in the ecology of certain nematodes. PMID- 19295842 TI - Electron Microscope Characterization of Carbohydrate Residues on the Body Wall of Xiphinema index. AB - The location of carbohydrate moieties on the outer cuticle of Xiphinema index was examined by electron microcopy using several different reagents: a) The periodic acid-thiosemicarbazide-silver proteinate reaction was used as a general stain for carbohydrates. In sectioned material it stained the canal system and deeper layers of the cuticle as well as the outer surface, b) Cationized ferritin at pH 2.5, which identifies carboxyl and sulfate groups, was used to identify sialic acid residues and also labelled parts of the canal system, c) Ferritin-goat anti rabbit IgG coupled to a DNP ligand was used to label either sialyl or galactosyl/N-acetyl-D-galactosaminyl residues, d) Ferritin hydrazide, a new reagent, was used for the ultrastructural localization of glyco-conjugates. Reagents c) (with appropriate antisera) and d) were applied only to the outer surfaces of the cuticle; they showed that sialic acid residues were concentrated mainly on the outer body wall of the head, the lips, oral opening, amphid apertures, and outer surface of protruded odontostyles. Ferritin distribution was not altered by pretreatment with neurantinidase. Galactose oxidase treatments revealed galactose/N-acetyl-D-galactosamine residues along the entire body wall. These results confirmed earlier findings obtained by fluorescence microscopy. PMID- 19295843 TI - Caenorhabditis elegans: Stage Specific Differences in Cuticle Surface Carbohydrates. AB - Stage-specific differences in wheat germ agglutinin (WGA) binding saccharides were demonstrated between the surfaces of the eggs, L1 larvae, young aduhs, and old adults of Caenorhabditis elegans. The WGA binding was to n-acetylglucosamine groups but not to terminally linked n-acetylneuraminic acids. An age-related decrease in WGA binding occurred in adults, supporting previous findings of a decrease in net negative cuticle surface charge during aging. PMID- 19295844 TI - Interrelationships between Ethylene Production, Gall Formation, and Root-knot Nematode Development in Tomato Plants Infected with Meloidogyne javanica. AB - Ethylene production was determined in excised tomato (Lycopersicon esculentum) root cultures of Meloidogyne javanica susceptible and resistant cultivars infected with M. javanica. Uninfected cultivars produced very low amounts of ethylene. Relatively high amounts of ethylene were produced by the infected susceptible cultivars. Peak production of 1.6 n moles * g root(1) * h(1) occurred between 9 and 16 days after inoculation (DAI). The period of high ethylene production coincided with that of rapid increase in gall weight. Low amounts of ethylene were also released by the infected resistant cultivar between 9 and 12 DAI, which follows the hypersensitivity reaction. Ethylene production in infected intact plants during the period of rapid gall growth was twice as much as in uninfected plants during the same time. Exposing excised root cultures to 0.5 or l0 ppm ethylene accelerated the rate of increase in gall weight of M. javanica infected roots. In contrast, overall root growth was inhibited by these treatments, compared to infected roots which were not exposed to ethylene. PMID- 19295845 TI - Host Response to Meloidodera spp. (Heteroderidae). AB - Host responses to Meloidodera floridensis Chitwood et al., 1956, M. charis Hooper, 1960, and M. belli Wouts, 1973 were examined on loblolly pine, peony, and sage, respectively, with light, scanning, and transmission electron microscopy. In each case the nematodes induce a single uninucleate giant cell. The giant cell is initiated in the pericycle and expands as it matures. The mature giant cell induced by M. floridensis is surrounded by vascular parenchyma, whereas that caused by M. charts and M. belli coutacts xylem and phloem. The cell wall of giant cells induced by all three Meloidodera spp. is generally thicker than that of surrounding cells, with the thickest part adjacent to the lip region of the nematode. The thinner portion of the wall includes numerous pit fields with plasmodesmata, but wall ingrowths were not detected in a thorough examination of the entire wall. The nucleus of a giant cell induced by M. goridensis is highly irregular in shape with deep invaginations, whereas those caused by M. charis and M. belli include a cluster of apparently interconnected nuclear units. Organelles, including mitochondria, endoplasmic reticulum, and plastids of giant cells caused by Meloidodera, are typical of those reported in host responses of other Heteroderidae. The formation of a single uninucleate giant cell by Meloidodera, Cryphodera, Hylonerna, and Sarisodera, but a syncytium by Atalodera and Heterodera sensu lato, might be considered in conjunction with additional characters to determine the most parsimonious pattern of phylogeny of Heteroderidae. PMID- 19295846 TI - Effect of Plant Age and Transplanting Damage on Sugar Beets infected by Heterodera schachtii. AB - Sugar beet (Beta vulgaris L. cv. Monogerm C.S.F. 1971) seeds sown into Vineland fine sandy loam, infested with 15,500 H. schachtii juveniles/pot, showed little growth during an 11-week test in the greenhouse. Seedlings transplanted at 2, 4, and 6 weeks of age had 32, 30, and 31% less top weight and 71, 68, and 59% less root weight, respectively, compared to controls grown in nematode-free soil. Nematode reproduction in both direct-seeded and transplanted sugar beets was limited and related to root weight. Shoot/root ratios were increased by the nematodes in all nematode-infected beets compared to those grown in soil without nematodes. In contrast to seeding or transplanting sugar beets into nematode infested Vineland fine sandy loam, an inoculation of Beverly fine sandy loam supporting 0 (seeds), 2-, 4-, and 6-week-old sugar beet seedlings with 7,400 juveniles/pot, followed by 11 weeks of growth in the growth-room, resulted in top weight losses of only 13, 3, 18, and 15% and losses in root weight of 44, 38, 36, and 38%, respectively. Nematode reproduction was high and all shoot/root ratios were increased by the nematode compared to the noninoculated controls. These experiments have shown that sugar beets sown into nematode-infested soil are damaged much more heavily by H. schachtii juveniles than seeds inoculated with the nematode immediately following sowing. Results indicate that an increase in tolerance of sugar beets to attack by H. schachtii does not occur beyond the first 2 weeks of growth and that transplanting damage lowers the tolerance of seedlings to nematode attack. PMID- 19295847 TI - Influence of Six Vegetable Cultivars on Reproduction of Meloidogyne javanica. AB - Replicated field and greenhouse experiments were used to evaluate the effect of tomato, cabbage, cucumber, carrot, Amaranthus hybridus, and pepper on growth and fecundity of Meloidogyne spp., particularly M. javanica. In the field tests, tomato, cucumber, and carrot favored population increases of Meloidogyne spp., while Amaranthus, pepper, and cabbage limited them. Some cropping sequences that included crops from the latter group had a suppressive effect on population growth. Thus, of the 36 cropping sequences that were investigated, the following kept the pests in check: tomato-pepper; tomato-Amaranthus; cabbage-pepper; Amaranthus-pepper; carrot-cabbage; pepper-pepper; pepper-Amaranthus; and Amaranthus-pepper. In the greenhouse tests, tomato, cucumber, and carrot had a high number of galls per 50 cm of root, large, conspicuous galls and egg masses, and a high number of larvae per egg mass. Thus, they were highly susceptible. Cabbage and Amaranthus were unsuitable hosts as reflected in the absence of galls or a low number per 50 cm of root. small size of galls and egg masses, and few progeny on the subsequent crop of pepper. The length of time required for eggs to hatch on different hosts varied considerably and is thought to be a significant factor in infection of hosts. PMID- 19295848 TI - Influence of Meloidogyne hapla Chitwood, 1949 on Development and Establishment of Heterodera schachtii Schmidt, 1871 on Beta vulgaris L. AB - Influence of Meloidogyne hapla on estahlishnrent and maturity of Heterodera schachtii in sugarbeet was studied. Results indicated that when the majority of M. hapla were in second, third, or fourth larval stages within plants prior to H. schachtii inoculation, growth and development of the latter was retarded. However, when M. hapla reached the young female stage prior to inoculation of H. schachtii, establishment and development of the latter was greatly enhanced. As M. hapla reached maturity before and after egg production prior to H. schachtii inoculation, establishment and growth of the latter was progressively decreased. In each instance, M. hapla developed independently and matured at the same rate as in plants inoculated with only M. hapla. Usually ratios of total soluble carbohydrates to reducing carbohydrates were lower, but not significantly different, in plants receiving both nematodes as compared to other treatments. PMID- 19295849 TI - Effect of an Extract from Saprozoic Nematode-Infested Compost on the Mycelial Growth of Agaricus brunnescens. AB - Extracts from compost infested with Caenorhabditis elegans suppressed mycelial growth of Agaricus brunnescens. An extract from uninfested compost also inhibited mycelial growth but to a lesser degree. The critical role of compost bacteria and/or other compost micro-organisms is implicated by these results. PMID- 19295850 TI - Biology of Heterodera mediterranea. AB - Heterodera mediterranea completes embryogenic development in 15-18 days at 24 +/- 2 C. On olive and pistachio seedlings the postembryogenic development was completed in 42-50 days at 24-30 C. Juveniles and adults have semiendoparasitic habits and do not penetrate completely into the root tissue. This cyst forming nematode has been detected only on Olea europaea, Pistacia lentiscus, and P. vera. Syncytia formation and disorder of root stelar structure are the main anatomical changes induced by the parasite. PMID- 19295851 TI - Reproductive Development of Verutus volvingentis (Tylenchida: Heteroderidae). AB - Verutus volvingentis Esser, 1981 deposits eggs in the rhizosphere without a gelatinous matrix. Ecdysis was not observed to occur in the egg. Spicular primordia in the rectal area of a second-stage larva were well defined. One larva increased in width from 28.2 mum to a maximum of 51.7 mum after 176.5 hours of feeding, prior to the second ecdysis. It then decreased steadily in width to 33.3 mum, at which time it had molted to a fully developed male. Males leave the third stage larval integument embedded in the root following final ecdysis. The unique feature of female development was the occurrence of large vaginal primordial cells. Male and female development took from 6 to 15 days and 17 days, respectively. PMID- 19295852 TI - Taxonomic Limits of the Genus Nagelus (Thorne and Malek, 1968 Siddiqi, 1979 with a Description of Nagelus borealis n.sp. from Alaska. AB - The genus Nagelus (Thorne and Malek, 1968) Siddiqi, 1979 is modified and a new species from Alaska is described. The combination of scanning electron microscopy and light microscopy permits the characterization of Nagelus spp. as having a broadly oval face pattern, no longitudinal striations on the lip region, deirids surrounded by six incisures, and an irregularly tapering tail with a large hyaline region. Nagelus leptus (Allen, 1955) Siddiqi, 1979, N. alpensis Doucet and Luc, 1981, N. camelliae (Kheiri, 1972) Siddiqi, 1979, N. jamelensis (Nesterov, 1973) Siddiqi, 1979, and N. obscurus (Allen, 1955) n. comb. are retained on this basis. Nagelus borealis n. sp. is characterized by a styler length of 30 mum or more, an irregularly scalloped perioral disc, and a proportionately larger basal bulb. Nagelus abalosi (Doucet, 1978) Doucet. 1980 and N. virginalis (Doucet, 1978) Doucet, 1980 are synonymized with N. leptus. Nine other species from Nagelus are transferred to Merlinius Siddiqi, 1970. PMID- 19295853 TI - Effect of Aedes aegypti (Diptera:Culicidae) Age on Sex Ratios in Romanomermis culicivorax (Nematoda:Mermithidae). AB - First, second, third, early and late fourth-instar larvae, and pupae of Aedes aegypti were infected with Romanomerrais culicivorax and reared at 20, 25, and 30 C. An increase in the ratio of male to female nematodes was observed with increase in host age at the time of infection at each temperature. The number of pupal and late fourth-instar infections was low, but R. culicivorax continued to develop in adult A. aegypti. Since male nematodes were recovered from both male and female hosts infected as late fourth instars or pupae, the sex of the host did not influence the sex of the nematode. PMID- 19295854 TI - Susceptibility of the Colorado Potato Beetle and the Sugarbeet Wireworm to Steinernema feltiae and S. glaseri. AB - In laboratory tests, larvae of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), and the sugarbeet wireworm (SBW), Limonius californicus (Mannerheim), were exposed to the nematodes Steinernema feltiae Filipjev (Mexican strain) (= Neoaplectana carpocapsae) and S. glaseri Steiner in soil. S. feltiae caused significantly higher mortality in SBW larvae than did S. glaseri, but both nematode species were equally effective against CPB larvae. The minimum concentration of S. feltiae for 100% mortality of CPB larvae after 13 days was 157 nematodes/cm(2) of soil, and the LC based on 6-day mortality was 47.5 nematodes/cm(2); in contrast, 100% mortality of SBW larvae was not achieved with even the highest concentration tested, 393 nematodes/cm(2). CPB adults emerging from nematode-contaminated soil were not infected. In field cage tests, S. feltiae applied to the soil surface at the rates of 155 and 310 nematodes/cm(2) soil caused 59% and 71% mortality, respectively, of late-fourth-instar spring generation CPB, and 28% and 29% mortality, respectively, of SBW. No infection was obtained when larvae of summer generation CPB and SBW were placed in the same cages approximately 6 weeks after nematodes were applied to the soil. Inundative soil applications of S. feltiae, though cost prohibitive at present, were effective in reducing caged CPB and SBW field populations. PMID- 19295855 TI - Characteristics of herbicide distribution as patterns for nematicide behavior. PMID- 19295856 TI - Progress and prospects in the chemotherapy of nematode infections of man and other animals. PMID- 19295857 TI - Insecticides and their design. PMID- 19295858 TI - Reproduction of Two Races of Meloidogyne incognita in Tomato Plants Grown at High Temperature. PMID- 19295859 TI - Monacrosporium Lysipagum Infecting Egg Masses of Meloidogyne acrita. PMID- 19295860 TI - Response of Two Alfalfa Cultivars to Meloidogyne hapla. PMID- 19295862 TI - Effects of Pangola,Digitgrass on Meloidogyne arenaria, M. javanica, and M. hapla. PMID- 19295861 TI - Root Extracts of Pangola Digitgrass Affect Egg Hatch and Larval Survival of Meloidogyne incognita. PMID- 19295863 TI - Isolation of an Unusual Entaphelenchid (Tylenchida: Entaphelenchidae) from the Hemocoel of Platysoma Punctigerum LeConte Coleoptera: Histeridae). PMID- 19295864 TI - Vertical Migration of Heterorhabditis bacteriophora and H. heliothidis (Nematoda: Heterorhabditidae) in Sandy Loam Soil. PMID- 19295865 TI - Nematode damage functions: the problems of experimental and sampling error. AB - The development and use of pest damage functions involves measurement and experimental errors associated with cultural, environmental, and distributional factors. Damage predictions are more valuable if considered with associated probability. Collapsing population densities into a geometric series of population classes allows a pseudo-replication removal of experimental and sampling error in damage function development. Recognition of the nature of sampling error for aggregated populations allows assessment of probability associated with the population estimate. The product of the probabilities incorporated in the damage function and in the population estimate provides a basis for risk analysis of the yield loss prediction and the ensuing management decision. PMID- 19295866 TI - Effect of the Entomogenous Nematode Nemplectana carpocapsae on the Tachinid Parasite Compsilura concinnata (Diptera: Tachinidae). AB - The entomogenous nematode Neoaplectana carpocapsae and its associated bacterium, Xenorhabdus nematophilus, could not infect the pupal stage of the tachinid Compsilura concinnata through the puparium. N. carpocapsae had an adverse effect on 1-, 2- and 3-day-old C. concinnata larvae within the armyworm host in petri dish tests. All 1-day-old larvae treated with nematodes died in their hosts, whereas 61% and 69% of 2- and 3-day-old larvae treated with nematodes, respectively, died. However, the survivors developed to adults. Nine to thirty seven percent of adult tachinids which emerged from nematode-treated soil (50 nematodes/cm(2)) were infected with N. carpocapsae. The nematode adversely affects C. concinnata directly by the frank infection of the tachinid and indirectly by causing the premature death of the host which results in tachinid death. PMID- 19295867 TI - Transport of Palmitic Acid Across the Tegument of the Entomophilic Nematode Romanomermis culicivorax. AB - Romanomermis culicivorax juveniles, dissected out of Aedes aegypti larvae 7 days after infection, were incubated under controlled conditions in isotonic saline containing (1)C-U-palmitic acid to investigate the nature of the transport mechanism(s) used by the nematode for transcuticular uptake of palmitic acid. Net uptake of the isotope by the nematode was of a logarithmic nature with respect to time. Uptake of palmitic acid was accomplished by a combination of diffusion and a mediated process which was substrate saturable and competitively inhibited by myristic and stearic acids. Both 2,4-dinitrophenol and ouabain inhibited uptake of palmitic acid and thus supported the hypothesis that the carrier system is of the active transport variety and is coupled to a NaK ATPase pump. PMID- 19295868 TI - In Vitro Hatch and Survival of Heterodera glycines as Affected by Alachlor and Phenamiphos. AB - Heterodera glycines (race 1) eggs were exposed to aqueous solutions o f selected concentrations o f the herbicide alachlor and the organophosphate nematicide phenamiphos alone and in herbicide-nematicide combinations. Phenamiphos (0.5 mug/ml) + alachlor (0.063, 0.125, or 1.0 mug/ ml) treatments increased the incidence o f juvenile hatch over that of untreated controls at 18 days. At 18 and 25 days, phenamiphos (0.5 mug/ml) treatments contained more juveniles than did phenamiphos at 1.0 mug/ml. Phenamiphos (1.0 mug/ml) alone and in combination with alachlor (1.0 mug/ ml) suppressed hatch for 21 days and juvenile survival for more than 21 days. Alachlor treatments enhanced juvenile survival compared to the untreated control at 14 and 21 days. Technical alachlor gave results similar to those of the formulated product. PMID- 19295869 TI - Activity and Survival of Orrina phyllobia: Preliminary Investigations on the Effects of Solutes. AB - The motility and survival of Orrina phyllobia fourth-stage juveniles (J4) were examined in NaCl, sucrose, and synthetic soil solutions. Synthetic soil solutions (SSSs) contained Na, K, Ca(2), Mg(2), Cl, and NO at relative concentrations identical to those in a known agronomic soil. Nematode activity was dependent on solute composition and on water potential. In all solutions, motility ceased at a water potential of -30 x 10 Pa and nematodes partially desiccated. Activity inhibition in NaCl began at -5 x 10 Pa. At -15 x 10 Pa, a high level of activity was sustained only in SSS. Lethal effects occurred at -15 and -60 x 10 Pa in NaCl and SSS, respectively. No lethal effects were measured in sucrose solutions. Hydrogen ion concentration over the pH range 4.5-9.5 had no measurable effect on nematode activity. PMID- 19295870 TI - Effects of the Ionic Composition and Water Potential of Aqueous Solution on the Activity and Survival of Orrina phyllobia. AB - The activity and survival of Orrina phyllobia fourth-stage juveniles (J4) were examined in aqueous solutions representing 96 combinations of eight predominant soil solution ions at total concentrations of 100, 200, and 1,000 meq/liter. Various water potentials were imposed by the addition of mannitol or polyethylene glycol to ionic solutions. Nematode longevity increased as water potential was decreased. Longevity was approximately doubled at a water potential of -23 x 10 Pa and more than tripled at -60 x 10 Pa. No combination oflons at 200 meq/liter was lethal after a 6-day exposure. Several ion combinations significantly increased longevity at -10 and -23 x 10 Pa. Single cation Na solutions consistently inhibited activity and more than doubled nematode longevity. PMID- 19295871 TI - Transmission of the Pinewood Nematode, Bursaphelenchus xylophilus, to Slash Pine Trees and Log Bolts by a Cerambycid Beetle, Monochamus titillator, in Florida. AB - Field-collected adults of the southern pine sawyer, Monochamus titillator (F.) (Coleoptera: Cerambycidae), naturally infested with fourth-stage juveniles (dauerlarvae) of the pinewood nematode, Bursaphelenchus xylophilus (Steiner and Buhrer, 1934) Nickle, 1970, were maturation fed on excised shoots of typical slash pine, Pinus elliottii Engelm. var elliottii, for 21 days. During August 1981, a male and female adult beetle were held in a sleeve cage placed on the terminal of a side branch of each of seven replicate, healthy 10-year-old slash pine trees. All seven branch terminals showed evidence of beetle feeding on the bark after 1 week, and pinewood nematodes were present in wood samples taken near these feeding sites. Four of the seven trees showed wilt symptoms in 4-6 weeks and died about 9 weeks after beetle feeding. Pinewood nematodes were recovered from the roots and trunks of the dead trees. Each of seven replicate slash pine log bolts was enclosed in a jar with a pair of the same beetles used in the sleeve cages. After 1 week, wood underlying beetle oviposition sites in the bark of all replicate log bolts was infested with the pinewood nematode. PMID- 19295872 TI - Early Growth of Soybean as Altered by Heterodera glycines, Phenamiphos and/or Alachlor. AB - Greenhouse and field experiments were conducted to determine the effects of phenamiphos and/or alachlor on early growth of soybean, root morphology, and infection and resurgence of Heterodera glycines (race 1). All tests were planted to 'Ransom' soybeans. In greenhouse experiments without nematodes, root growth was inhibited at 5 days by alachlor treatments and at 10 days by phenamiphos treatments; with nematodes, phenamiphos treatments enhanced root growth. Phenamiphos also suppressed early penetration of soybean roots by H. glycines in the greenhouse. Early soybean growth parameters among treatments were generally similar in the field. Nematode penetration was limited with treatments containing phenamiphos at one location. Plants treated with only alachlor had less nematode infection than did the control; however, plants treated with herbicide/nematicide combinations had more nematode penetration than did plants treated with phenamiphos alone. Alterations of root growth and interference with the efficacy of phenamiphos are two processes by which alachlor may enhance soybean susceptibility or suitability to H. glycines. PMID- 19295873 TI - Nematode Population Attrition and Histopathology of Heterodera glycines-Soybean Associations. AB - Selected populations of soybean cyst nematodes were inoculated to roots of compatible and incompatible soybeans. Rates of penetration of infective juveniles of nematode populations selected on PI 209332, PI 89772, and Pickett 71 were equivalent on compatible and incompatible soybean roots. The first two populations averaged about 10% and the last about 5% penetration in 24-hour inoculations of young seedlings. About 14% of those juveniles that entered roots in compatible combinations developed into maturing females, compared with only about 1% in incompatible combinations. Several aberrations from the pattern of syncytial development associated with mature females in compatible hosts were apparent. A rapid necrotic response occurred in both kinds of hosts but was more frequent in incompatible associations. Delayed necrosis and small syncytia were present in some combinations. Those few females that developed in incompatible soybeans were associated with a characteristic syncytium different from the kind seen in roots of compatible hosts. PMID- 19295874 TI - Phytotoxin Production in Bursaphelenchus xylophilus-Infected Pinus sylvestris. AB - Our findings suggest that i) phytotoxic materials can be isolated from Bursaphelenchus xylophilus-infected Scots pine, but not from noninfected pines; ii) the phytotoxins cause wilting of 45-day-old and 2-year-old pine seedlings in a dose and species dependent manner; iii) the phytotoxins are produced early in the infection, accumulate or increase with time, and may function to suppress water transport in the tree; and iv) the phytotoxins are lipid materials of low molecular weight which are not acidic. PMID- 19295875 TI - Pterotylenchus cecidogenus n. gen., n. sp., a New Stem-gall Nematode Parasitizing Desmodium ovalifolium in Colombia. AB - Pterotylenchus cecidogenus n. gen., n. sp. from stem-galls of Desmodium ovalifolium Wall. in Colombia is described and illustrated. The new genus belongs to Anguinidae and is related to Orrina Brzeski, 1980 but is unique in having large vulval flaps. P. cecidogenus has females with body 0.59-0.8 mm long, stylet 9-11 mum long, no median oesophageal bulb, a crustaformeria of 32-36 cells, a short post-vulval uterine sac, and a conical pointed tail. PMID- 19295876 TI - Occurrence of Aphelenchoides besseyi in Louisiana Rice Seed and Its Interaction with Sclerotium oryzae in Selected Cultivars. AB - Aphelenchoides besseyi, the nematode causal agent of white-tip disease of rice, was recovered from 5.5% of 474 seed samples obtained from rice seed warehouses in Louisiana. Laboratory tests in which A. besseyi-infested rice seed was treated with Phostoxin(R), a compound used for control of insects in stored grain, indicate that it also has nematicidal properties. In 18-week-duration greenhouse tests, populations of A. besseyi increased 4-5-fold on the cultivars Saturn and Melrose and 3-fold on Nova '76. Green weights of Nova '76 plants inoculated with A. besseyi and Sclerotium oryzae, the causal agent of rice stem rot, were significantly reduced below those of plants inoculated with either organism alone or with distilled water. Weights of Melrose plants were reduced significantly by treatments with A. besseyi alone and A. besseyi plus S. oryzae, but not by S. oryzae alone. Saturn plant weights were not reduced significantly by either organism alone or by the two in combination. PMID- 19295877 TI - Histopathology and Host Range Studies of the Redwood Nematode Rhizonema sequoiae. AB - Second-stage larvae of Rehizonma sequoiae Cid del Prado Vera et al. tunnel through the cortex of the redwood Sequoia sempervirens (D. Don) Endl. root to the vascular tissue where each developing female induces a single ovoid or occasionally spherical giant cell with a single ovoid to spherical nucleus containing one to four enlarged nucleoli. Nematode tunnels are filled with a gel material and often contain second-stage larvae and males. There is tissue necrosis around females, and cortical tissue is destroyed after infection by many second-stage larvae. R. sequoiae females developed to maturity on S. sempervirens, Acer macrophyllum Pursh, AInus rhombifolia Nutt., Libocedrus decurrens Torr, Pseudotsuga menziesii (Mirb.) Franco, and Sequoiadendron giganteum (Lindl.) Decne. In the Marin County, California, forest mature females were also found naturally infecting Lithocarpus densiflorus (Hook &Arn.) Rehd., Umbellularia californica (Hook &Arn.) Nutt., and Arbutus menziesii Pursh. PMID- 19295878 TI - Postembryonic Development of the Redwood Nematode, Rhizonema sequoiae (Nemata: Heteroderidae). AB - Second-stage larvae of Rhizonema sequoiae Cid del Prado Vera et al. developed into adult females in 6 months or adult males in 3 - 4 months on roots of Sequoia sempervirens maintained in a growth chamber at 16 C with a 12-hour light period. Under these conditions the second-stage larvae increased in diameter, the central cells of the genital primordium increased in size, and their nuclei enlarged. Mesenchymal cells accumulated in the esophageal and tail regions. Second-stage larvae become third-stage males or females 2 months after inoculation of redwood roots. Their sex could be distinguished by the ratio of length to width of the genital primordium, 3.4 for males and 1.6 for females. The stylet in both sexes became slender, the median bulb became robust and almost spherical, and rings of punctation on the cuticle were evident. Fourth-stage females developed in 3 months from the time of inoculation, and fourth-stage males in slightly less time. At this stage the females were more swollen than the males, the rectum was conspicuous, their reproductive system was in the process of elongation, and the annulation of the cuticle was more evident. The ratio of males to females was 2.3. Mature females were completely inside the roots and did not form cysts. The cuticle was entirely annulated, and the first eggs were detected inside the female 4 months after inoculation and started the production of abundant gelatin like material. The new generation of second-stage larvae hatched inside the female 2 months after she matured, completing the life cycle in 8 months. The redwood nematode also completed its life cycle in 8 months under greenhouse conditions, but the ratio of males to females increased to 7.4. The entire nematode population died out at 25 C after 6 months. In a Marin County, California, forest, where this nematode occurs naturally, the temperature averaged only 9 C over the November to June period of this study, and the redwood nematode reached the fourth stage with a male-to-female ratio of 1.8. PMID- 19295879 TI - Description of Males of Heterodera zeae. AB - The male ofHeterodera zeae, the corn cyst nematode, is described and illustrated for the first time. Specimens were obtained from a culture originating from cysts collected in Kent County, Maryland, at the site of the first known infestation of H. zeae in the United States. PMID- 19295880 TI - Some Factors Affecting Survival of Desiccation by Infective Juveniles of Orrina phyllobia. AB - The survival of desiccation by J4 Orrina phyllobia was examined at controlled relative humidities. When nematodes were transferred from water to air at 10% relative humidity (rh), 80% died within 30 minutes. When nematodes were transferred from water to air with rh at 70% or greater for ca. 15 minutes prior to being transferred to 10% rh, more than 90% of them survived desiccation. This phenomenon is referred to as preconditioning and occurred at much faster rates (2 30 minutes) than has been observed for other nematode species (24 hours). Differences in preconditioning rates may be due to technique-dependent variations in boundary layer resistance around nematodes during desiccation. PMID- 19295881 TI - Application of Isoelectric Focusing to the Taxonomic Identification of Meloidogyne spp. AB - Meloidogyne incognita, M. arenaria, M. hapla, and M. javanica were distinguishable from each other by isoelectric focusing (IEF) of nematode egg proteins. Proteins extracted from larvae and adults of Hoplolaimus columbus and from eggs of Heterodera glycines had distinctive profiles, also. Protein profiles from eggs, preparasitic larvae and egg-laying adults of M. incognita showed differences. It was necessary to compare samples run at the same time to ensure reliability. PMID- 19295882 TI - Nodulation of Soybeans as Affected by Half-root Infection with Heterodera glycines. AB - A split-root technique was applied to soybean, Glycine max (L.) Merr. cv. Lee 68, to characterize the nature of the nodulation suppression by race 1 of the soybean cyst nematode (SCN), Heterodera glycines. Root-halves of each split-root plant were inoculated with Rhizobium japonicum, and one root-half only was inoculated with various numbers of SCN eggs. Nodulation (indicated by nodule number, nodule weights, and ratio of nodule weight to root weight) and nitrogen-fixing capacity (indicated by rate of acetylene reduction) were systemically and variously suppressed on both root-halves of the split-root plant 5 weeks after half-root inoculation with 12,500 SCN eggs. Inoculation with 500 eggs caused this suppression only on the SCN-infected (+NE) root-half; nodulation on the companion uninfected (-NE) root-half was stimulated slightly. The +NE root-halves inoculated with 5,000 eggs were excised at 2-week intervals; nodulation on the remaining -NE root-halves was not different from that of the noninoculated control when measured 6 weeks after the SCN inoculation. Thus, the systemic suppression of nodulation was reversible upon the removal of the SCN. Similarly, application of various levels of KNO to the -NE root-halves of the split-root plant did not alleviate the suppressed nodulation on the companion +NE root halves, even though plant growth was much improved at certain levels of nitrogen (125 mug N/g soil). This indicated that the localized suppression of nodulation by SCN was caused by factors in addition to poor plant growth. PMID- 19295883 TI - Histopathology of Okra and Ridgeseed Spurge Infected with Meloidodera charis. AB - Histological observations of okra Abelomoschus esculentus 'Clemson Spineless' and ridgeseed spurge Euphorbia glyptosperma (a common weed) infected with Meloidodera charis Hopper, indicated that the juvenile nematode penetrated the roots intercellularly. Within 5 days after plant emergence the nematode positioned its body in the cortical tissue parallel to the vascular system. By 10 days after plant emergence the juvenile had extended its head into the vascular system and initiated giant cell formation, generally in protophloem tissue. Giant cells were one celled and usually multi-nucleate. Eggs were observed in the female body 30 days after plants emerged and juveniles were found within the female body by 40 days. Nematode development progressed equally in the root system of either host plant. Generally, throughout the nematode's life cycle its entire body remained inside the cortical tissue of okra. In ridgeseed spurge, however, the posterior portion of the female erupted through the host epidermis as early as 15 days after plant emergence; only the head and neck remained embedded in the host. The nematode caused extensive tissue disruption in the cortical and vascular system of both plant species. Corn, Zea mays, was another host of the nematode. PMID- 19295885 TI - An Observation Chamber Technique for Evaluating Potential Biocontrol Agents of Globodera rostochiensis. PMID- 19295884 TI - Inhibitory Effect of Watermelon Mosaic Virus on Meloidogyne javanica (Treub) Chitwood Infecting Cucurbita pepo L. PMID- 19295886 TI - Seasonal Variation of a Meloidogyne exigua Population in a Coffee Plantation. PMID- 19295887 TI - Anhydrobiosis in five species of plant associated nematodes. AB - Five species of nematodes - Hemicriconemoides pseudobrachyurum, Hemicycliophora conida, Macroposthonia ornata, Aphelenchoides ritzemabosi, and Psilenchus hilarulus -were desiccated to study their capacity to survive anhydrobiotically. Results indicate that the ability of the sheath to shrink quickly and its relatively loose attachment with the nematode body allow H. conida to survive longer than H. pseudobrachyurum; the survival of M. ornata was intermediate, A. ritzemabosi and P. hilarulus survived immersion in paraffin oil for 12 and 17 days, respectively. Both of these nematodes possess multiple contraction ability; i.e., coiling coupled with transverse and longitudinal folding of the cuticle. P. hilarulus is a new addition to the list of anhydrobiotic nematodes. PMID- 19295888 TI - Influence of Urea, Hydroxyurea, and Thiourea on Meloidogyne javanica and Infected Excised Tomato Roots in Culture. AB - Urea (U), hydroxyurea (HU), and thiourea (TU), in various concentrations, were added to chemically defined plant tissue culture medium on which Meloidogyne javanica was reared on excised tomato roots. Concentrations as low as 3 ppm HU or 12 ppm TU inhibited nematode maturation by 70-90% 4 weeks after inoculation, and the coenocytes in the parasitized tissue were poorly developed. Gall weight was also inhibited by 50% in cultures treated with 3 and 6 ppm HU. However, exposing juveniles of M. javanica and Tylenchulus semipenetrans or juveniles and adults of Pratylenchus thornei to increasing concentrations of HU or TU, up to 100 ppm, was not lethal. These two urea derivatives still inhibited nematode maturation when the infected region of the root was not in direct contact with the chemicals. Therefore, we suggest that these urea derivatives inhibit nematode development by affecting the plant metabolism essential to coenocyte formation, an occurrence similar to the hypersensitive reaction in a naturally resistant plant. PMID- 19295889 TI - Protocylindrocorpus dendrophilus n. sp. (Nematoda: Cylindrocorpidae) Associated with Pine Wood Borings. AB - Protocylindrocorpus dendrophilus n. sp. is described from xylem samples taken from beetle infested slash pine (Pinus elliottii Engelmann var. elliottii) in Central Louisiana. It is similar to P. goodeyi (Ruhm) Paramonov, but differs by the possession of a protuberant and more posteriorly located vulva and in the position of the caudal papillae. Morphometrics of the male and female are presented. PMID- 19295890 TI - Relationship Between Heterodera schachtii, Meloidogyne hapla, and Nacobbus aberrans on Sugarbeet. AB - Heterodera schachtii, Meloidogyne hapla, and Nacobbus aberrans either alone, or in various combinations with each other, can, when inoculated at a concentration of 12 second-stage juveniles/ cm(3) of soil, cause a significant (P = 0.01) suppression of growth of sugarbeet (cv. Tasco AH14) seedlings. M. hapla and H. schachtii decreased growth of sugarbeet more than N. aberrans over a 60-day period. The adverse effect of N. aberrans on the final population/initial population (Pf/Pi) ratio for either M. hapla or H. schachtii was dependent on time, and was more accentuated on that of M. hapla than on that of H. schachtii. Neither M. hapla nor H. schachtii had an adverse effect on the Pf/ Pi ratio of N. aberrans. N. aberrans is considered to be less aggressive on sugarbeet than either H. schachtii or M. hapla. Sections of sugarbeet roots infected simultaneously with H. schachtii and N. aberrans showed scattered vascular elements between the N. aberrans syncytium located in the central part of the root and that of H. schachtii in the peripheral position. PMID- 19295891 TI - Pathogenicity of Criconemella curvata to Alfalfa. AB - Large populations of Criconemella curvata and extended experimental periods were required to adversely affect yield of 'Moapa 69' alfalfa. C. curvata ectoparasitic feeding caused a reduction in feeder root numbers and tap root size and small lesions on tap and secondary roots. Greater reproduction occurred at 27 C than at 22 C, but the effect of the nematode on alfalfa growth was the same at both temperatures. PMID- 19295892 TI - Resistant Germplasm in Gossypium Species and Related Plants to Rotylenchulus reniformis. AB - Gossypium hirsutum, G. herbaceum, G. arboreum, G. barbadense, wild Gossypium spp., Hibiscus spp, and other Malvaceae were tested in the greenhouse to identify germplasm resistant to Rotylenchulus reniformis (Rr). Host resistance was based on Rr egg production per gram of root compared with known G. hirsutum susceptible 'Deltapine 16' as check. G. longicalyx and Sida rhombifolia were nonhosts. High levels of resistance were found in G. stocksii, G. somalense, and G. barbadense 'Texas 110.' Other cotton lines with potential value in breeding for Rr resistance were G. herbaceum P.I. 408775; G. arboreum P.I. 41895, P.I, 417891, CB 3839; and G. hirsutum 893. All these supported less than 20% of the egg production on the check. Seventy-three percent of the Hibiscus spp. tested were resistant. Female development and egg production reflected host resistance; healthy females and large egg masses were observed on susceptible plants, and degenerated females and small egg masses on resistant plants. Females penetrating nonhost G. longicalyx never matured to kidney shape. PMID- 19295893 TI - Nematicides and nonconventional soil amendments in the management of root-knot nematode on cotton. AB - Granular and liquid commercial humates, with micronutrients, and a microbial fermentation product were compared in several combinations with nematicides for their effects on cotton lint yield and root-knot nematode suppression. Fumigant nematicides effectively reduced cotton root galling caused by root-knot nematodes, and cotton lint yields increased. Organophosphates and carbamates were not effective. Occasionally, cotton lint yields were increased or maintained with combination treatments o f humates, micronutrients, and a microbial fermentation product, but galling o f cotton roots by root-knot nematodes was usually not reduced by these treatments. PMID- 19295894 TI - Control of Meloidogyne spp. on Russet Burbank Potato by Applying Metham Sodium through Center Pivot Irrigation Systems. AB - Metham sodium applied in October through center pivot irrigation systems was evaluated for control of Meloidogyne hapla at 374, 468, and 701 liters/ha and for control of M. chitwoodi at 468 liters/ha on potato. Metham sodium at the high rates effectively controlled M. hapla. No females were detected in the tubers at the high rates of nematicide application, whereas a mean of 19 and 69% of the tubers were infected at the low rate and in the nontreated controls, respectively. In the M. chitwoodi trial only 1.5% of the tubers in the treated plots were infected compared with 82% in the nontreated plots. Metham sodium effectively controlled M. chitwoodi to soil depths of 30, 61, and 91 cm. PMID- 19295895 TI - Influence of Meloidogyne chitwoodi and M. hapla on Wheat Growth. AB - Meloidogyne chitwoodi reduced the growth of winter wheat 'Nugaines' directly in relation to nematode density in the greenhouse, The relationship between top dry weight and initial nematode density suggests a tolerance limit of Nugaines wheat to M. chitwoodi of between 0.03 and 0.18 eggs/cm(3) of soil; the value for relative minimum plant top weight was 0.45 g and 0.75 g, respectively. Growth of wheat in field microplots containing four population densities (0.003, 0.05, 0.75 and 9 eggs/cm(3) soil) was not affected significantly at any inoculum level compared to controls during September to July, However, suppression of head weights of 'Fielder' spring wheat grown May-July occurred in microplots initially infested with 0.75 and 9 eggs/cm(3) soil. Reproduction (Pf/Pi) was poorer at these two inoculum levels as compared to the lower densities. In another greenhouse experiment, roots of wheat cultivars Fielder, 'Fieldwin,' 'Gaines,' 'Hyslop,' and Nugaines became infected by M. chitwoodi, but not by M. hapla. Reproduction of M. chitwoodi was less on Gaines and Nugaines than on Fielder, Fieldwin, or Hyslop. PMID- 19295896 TI - Reactions of Grape Rootstocks to Pratylenchus vulnus and Meloidogyne spp. AB - Five grape rootstocks were inoculated with 0, 100, 1,000, and 10,000 Pratylenchus vulnus. Dogridge and Saltcreek supported low average total numbers of P. vulnus, 136-705/pot, at 12 months after inoculation. Growth of both rootstocks was not affected. Harmony, Couderc 1613, and Ganzin 1 supported high average total numbers, 6-856 times the inoculum levels. Numbers in Harmony continued to increase at all levels but reduced root weight only at the 10,000 level after 12 months. Numbers in Couderc 1613 decreased by 15-30% after 12 months, and root weight was reduced at the 10,000 level. In Ganzin 1, total nematode numbers diminished after 12 months but were still at high levels; growth reduction was proportional to numbers of nematodes added. Meloidogyne incognita, M. javanica, and M. arenaria produced galls and egg masses in Harmony and Couderc 1613 only at 36 C. Galling in Ganzin 1 increased with increasing temperature. Galls in Ganzin 1 at 18 C supported mature females after 90 days. Harmony was resistant to M. incognita in single and concomitant inoculations of P. vulnus and M. incognita. At 250 days after inoculation, total numbers of P. vulnus increased above the inoculum level and the 150-day values; increase was greatest in P. vulnus added singly. Neither nematode species affected growth of Harmony. PMID- 19295897 TI - Comparative Morphometrics of Eggs and Second-Stage Juveniles of Heterodera schachtii and a Race of H. trifolii Parasitic on Sugarbeet in The Netherlands. AB - Measurements of second-stage juveniles of Heterodera schachtii from California and The Netherlands and a race of H. trifolii from The Netherlands were obtained and compared to determine if these populations can be differentiated by morphometrics. Juvenile lengths of 10 specimens from each of 10 cysts of each population were measured. Dimensions of tail regions of 20 juveniles from individual cysts of H. schachtii (California) and a like number of juveniles of H. trifolii (The Netherlands) were also obtained. The mean lengths of juveniles of H. schachtii from California and The Netherlands were not significantly different, but similar measurements of H. schachtii and H. trifolii were different (P = 0.05). Mean dimensions of tail lengths, tail widths, tail hyaline lengths, and tail length/tail width were significantly greater for H. trifolii than for H. schachtii. Also, dimensions of eggs of H. trifolii were significantly greater than dimensions of H. schachtii eggs. The investigations established that H. schachtii can be readily differentiated from H. trifolii by morphometrics of eggs and juveniles, Minimum sample sizes required for specified confidence intervals for each criterion measured are provided. PMID- 19295898 TI - Distributed Egg Production Functions for Meloidogyne arenaria in Grape Varieties and Consideration of the Mechanistic Relationship between Plant and Parasite. AB - Nematode egg production rates, as mediated by environmental conditions and host status, are important determinants of population development. Rates of egg production by Meloidogyne arenaria varied from 0.48 to 1.0 egg per female per DD (degree days above 10 C) in different grape varieties. The length of the egg production period ranged from 550 to 855 DD where measurable, and was generally longer in those varieties where the production rate was slow. We hypothesize that if a successful infection site is established, a constant number of eggs is produced if favorable environmental conditions prevail. Mechanistic coupling structures between plant growth and nematode population models are formulated. The nematode population influences metabolite supply through its effect on physiological efficiency and also acts as a metabolic sink; the degree of plant physiological stress influences nematode population development by affecting the sex ratio and egg production rates. PMID- 19295899 TI - Meloidogyne microcephala n. sp. (Meloidogynidae), a Root-knot Nematode from Thailand. AB - Meloidogyne microcephala n. sp. is described and illustrated from specimens obtained from tobacco (Nicotiana tabacum L.) in Thailand. The female perineal pattern usually has a low dorsal arch, coarse striae, and a series of small cuticular flaps around the tail terminus. The stylet of the female is 14.4 mum long, with large, square to rectangular stylet knobs, The distinctive male head region is narrow, small, and truncate with a low, flattened head cap. The stylet length is 20.6 mum, and the knobs are small, angular, and set off from the shaft. Mean length of second-stage juveniles is 457.5 mum, and stylet length is 9.3 mum. The tail tip in the juveniles is set off from the rest of the tail as a small finger-like projection. M. microcephala reproduces by mitotic parthenogenesis, and has a chromosome number of 2n = 36. PMID- 19295900 TI - Hoplolaimoidea (nematoda: tylenchida) from the aleutian islands with descriptions of four new species. AB - Four new species of hoplolaimoid nematodes (Merlinius adakensis, Pratylenchoides megalobatus, Pratylenchus pratensisobrinus, and Pratylenchus ventroprojectus) are described from Adak Island in the Aleutian chain. M. adakensis n. sp. is separated from other species by body length (0.96-1.3 mm), stylet length (32-36 mum), number of tail annules (49-68), and c' (3.1-4.1). P. megalobatus n. sp. differs from all known Pratylenchoides spp. by having a very long esophageal gland lobe (b' = 2.4-3.3, overlap 3-6 times the body width). P. pratensisobrinus n. sp. closely resembles P. pratensis (de Man) Filipjev, but has a longer stylet (15-17 mum), a longer tail (c = 12-15; c' = 2.8-3.7), and more tail annules (23 37). P. ventroprojectus n. sp. is distinguished by body length (392-475 mum), three lip annules, low and flattened cephalic capsule, and presence of terminal subventral projection. Pratylenchoides variabilis Sher, Helicotylenchus amplius Anderson &Eveleigh, and H. spitsbergensis Loof are also reported from Adak and Amchitka Islands. PMID- 19295901 TI - Redescription of Xiphinema americanum Cobb, 1913 with Comments on Its Morphometric Variations. AB - Xiphinema americanum is redescribed and illustrated from material collected from Arlington Cemetery, near Cobb's type locality (Falls Church, Virginia), Morphometric data showing variations within this species are given for three additional populations. PMID- 19295902 TI - A New Pathotype of Root-knot Nematode on Grape Rootstocks. PMID- 19295903 TI - Nematode persistence after fumigation: a methodological problem. PMID- 19295904 TI - Erratum. AB - [This corrects the article on p. 649 in vol. 15.]. PMID- 19295905 TI - Organocatalytic Highly Enantioselective Tandem Michael-Knoevenagel Reaction for the Synthesis of Substituted Thiochromanes. AB - Enantioenriched tetrasubstituted thiochromanes have been synthesized using a tandem Michael addition-Knoevenagel reaction between 2-mercaptobenzaldehydes and benzylidenemalonates with a 9-epi-aminoquinine thiourea derivative as the catalyst. Steric and electron effects were found to affect profoundly the enantioselectivity and diastereoselectivity of the reaction. PMID- 19295906 TI - Synthesis of hindered biphenyls by sequential non-transition metal-catalyzed reaction/palladium-catalyzed cross-couplings. AB - The sequential reaction of 1,2-dihalobenzenes with aryl lithiums followed by palladium-catalyzed cross-coupling reactions with Grignard reagents and arylboronic acids is described. This sequential reaction provides a convenient and expeditious access to tri-ortho substituted biaryl derivatives. PMID- 19295907 TI - Three new structures of left-handed RADA helical filaments: structural flexibility of N-terminal domain is critical for recombinase activity. AB - RecA family proteins, including bacterial RecA, archaeal RadA, and eukaryotic Dmc1 and Rad51, mediate homologous recombination, a reaction essential for maintaining genome integrity. In the presence of ATP, these proteins bind a single-strand DNA to form a right-handed nucleoprotein filament, which catalyzes pairing and strand exchange with a homologous double-stranded DNA (dsDNA), by as yet unknown mechanisms. We recently reported a structure of RadA left-handed helical filament, and here present three new structures of RadA left-handed helical filaments. Comparative structural analysis between different RadA/Rad51 helical filaments reveals that the N-terminal domain (NTD) of RadA/Rad51, implicated in dsDNA binding, is highly flexible. We identify a hinge region between NTD and polymerization motif as responsible for rigid body movement of NTD. Mutant analysis further confirms that structural flexibility of NTD is essential for RadA's recombinase activity. These results support our previous hypothesis that ATP-dependent axial rotation of RadA nucleoprotein helical filament promotes homologous recombination. PMID- 19295908 TI - Is the new primate genus rungwecebus a baboon? AB - BACKGROUND: In 2005, a new primate species from Tanzania, the kipunji, was described and recognized as a member of the mangabey genus Lophocebus. However, molecular investigations based upon a number of papionins, including a limited sample of baboons of mainly unknown geographic origin, identified the kipunji as a sister taxon to Papio and not as a member of Lophocebus. Accordingly, the kipunji was separated into its own monotypic genus, Rungwecebus. METHODOLOGY/PRINCIPAL FINDINGS: We compare available mitochondrial and nuclear sequence data from the voucher specimen of Rungwecebus to other papionin lineages, including a set of geographically proximal (parapatric) baboon samples. Based on mitochondrial sequence data the kipunji clusters with baboon lineages that lie nearest to it geographically, i.e. populations of yellow and chacma baboons from south-eastern Africa, and thus does not represent a sister taxon to Papio. Nuclear data support a Papio+Rungwecebus clade, but it remains questionable whether Rungwecebus represents a sister taxon to Papio, or whether it is nested within the genus as depicted by the mitochondrial phylogeny. CONCLUSIONS/SIGNIFICANCE: Our study clearly supports a close relationship between Rungwecebus and Papio and might indicate that the kipunji is congeneric with baboon species. However, due to its morphological and ecological uniqueness Rungwecebus more likely represents a sister lineage to Papio and experienced later introgressive hybridization. Presumably, male (proto-)kipunjis reproduced with sympatric female baboons. Subsequent backcrossing of the hybrids with kipunjis would have resulted in a population with a nuclear kipunji genome, but which retained the yellow/chacma baboon mitochondrial genome. Since only one kipunji specimen was studied, it remains unclear whether all members of the new genus have been impacted by intergeneric introgression or rather only some populations. Further studies with additional Rungwecebus samples are necessary to elucidate the complete evolutionary history of this newly-described primate genus. PMID- 19295909 TI - Orally available selective melanocortin-4 receptor antagonists stimulate food intake and reduce cancer-induced cachexia in mice. AB - BACKGROUND: Cachexia is among the most debilitating and life-threatening aspects of cancer. It represents a metabolic syndrome affecting essential functional circuits involved in the regulation of homeostasis, and includes anorexia, fat and muscle tissue wasting. The anorexigenic peptide alpha-MSH is believed to be crucially involved in the normal and pathologic regulation of food intake. It was speculated that blockade of its central physiological target, the melanocortin (MC)-4 receptor, might provide a promising anti-cachexia treatment strategy. This idea is supported by the fact that in animal studies, agouti-related protein (AgRP), the endogenous inverse agonist at the MC-4 receptor, was found to affect two hallmark features of cachexia, i.e. to increase food intake and to reduce energy expenditure. METHODOLOGY/PRINCIPAL FINDINGS: SNT207707 and SNT209858 are two recently discovered, non peptidic, chemically unrelated, orally active MC-4 receptor antagonists penetrating the blood brain barrier. Both compounds were found to distinctly increase food intake in healthy mice. Moreover, in mice subcutaneously implanted with C26 adenocarcinoma cells, repeated oral administration (starting the day after tumor implantation) of each of the two compounds almost completely prevented tumor induced weight loss, and diminished loss of lean body mass and fat mass. CONCLUSIONS/SIGNIFICANCE: In contrast to the previously reported peptidic and small molecule MC-4 antagonists, the compounds described here work by the oral administration route. Orally active compounds might offer a considerable advantage for the treatment of cachexia patients. PMID- 19295910 TI - The Set2/Rpd3S pathway suppresses cryptic transcription without regard to gene length or transcription frequency. AB - In cells lacking the histone methyltransferase Set2, initiation of RNA polymerase II transcription occurs inappropriately within the protein-coding regions of genes, rather than being restricted to the proximal promoter. It was previously reported that this "cryptic" transcription occurs preferentially in long genes, and in genes that are infrequently transcribed. Here, we mapped the transcripts produced in an S. cerevisiae strain lacking Set2, and applied rigorous statistical methods to identify sites of cryptic transcription at high resolution. We find that suppression of cryptic transcription occurs independent of gene length or transcriptional frequency. Our conclusions differ with those reported previously because we obtained a higher-resolution dataset, we accounted for the fact that gene length and transcriptional frequency are not independent variables, and we accounted for several ascertainment biases that make cryptic transcription easier to detect in long, infrequently transcribed genes. These new results and conclusions have implications for many commonly used genomic analysis approaches, and for the evolution of high-fidelity RNA polymerase II transcriptional initiation in eukaryotes. PMID- 19295911 TI - Comparative transcriptional profiling of Bacillus cereus sensu lato strains during growth in CO2-bicarbonate and aerobic atmospheres. AB - BACKGROUND: Bacillus species are spore-forming bacteria that are ubiquitous in the environment and display a range of virulent and avirulent phenotypes. This range is particularly evident in the Bacillus cereus sensu lato group; where closely related strains cause anthrax, food-borne illnesses, and pneumonia, but can also be non-pathogenic. Although much of this phenotypic range can be attributed to the presence or absence of a few key virulence factors, there are other virulence-associated loci that are conserved throughout the B. cereus group, and we hypothesized that these genes may be regulated differently in pathogenic and non-pathogenic strains. METHODOLOGY/PRINCIPAL FINDINGS: Here we report transcriptional profiles of three closely related but phenotypically unique members of the Bacillus cereus group--a pneumonia-causing B. cereus strain (G9241), an attenuated strain of B. anthracis (Sterne 34F(2)), and an avirulent B. cereus strain (10987)--during exponential growth in two distinct atmospheric environments: 14% CO(2)/bicarbonate and ambient air. We show that the disease causing Bacillus strains undergo more distinctive transcriptional changes between the two environments, and that the expression of plasmid-encoded virulence genes was increased exclusively in the CO(2) environment. We observed a core of conserved metabolic genes that were differentially expressed in all three strains in both conditions. Additionally, the expression profiles of putative virulence genes in G9241 suggest that this strain, unlike Bacillus anthracis, may regulate gene expression with both PlcR and AtxA transcriptional regulators, each acting in a different environment. CONCLUSIONS/SIGNIFICANCE: We have shown that homologous and even identical genes within the genomes of three closely related members of the B. cereus sensu lato group are in some instances regulated very differently, and that these differences can have important implications for virulence. This study provides insights into the evolution of the B. cereus group, and highlights the importance of looking beyond differences in gene content in comparative genomics studies. PMID- 19295913 TI - Myostatin inhibition in muscle, but not adipose tissue, decreases fat mass and improves insulin sensitivity. AB - Myostatin (Mstn) is a secreted growth factor expressed in skeletal muscle and adipose tissue that negatively regulates skeletal muscle mass. Mstn(-/-) mice have a dramatic increase in muscle mass, reduction in fat mass, and resistance to diet-induced and genetic obesity. To determine how Mstn deletion causes reduced adiposity and resistance to obesity, we analyzed substrate utilization and insulin sensitivity in Mstn(-/-) mice fed a standard chow. Despite reduced lipid oxidation in skeletal muscle, Mstn(-/-) mice had no change in the rate of whole body lipid oxidation. In contrast, Mstn(-/-) mice had increased glucose utilization and insulin sensitivity as measured by indirect calorimetry, glucose and insulin tolerance tests, and hyperinsulinemic-euglycemic clamp. To determine whether these metabolic effects were due primarily to the loss of myostatin signaling in muscle or adipose tissue, we compared two transgenic mouse lines carrying a dominant negative activin IIB receptor expressed specifically in adipocytes or skeletal muscle. We found that inhibition of myostatin signaling in adipose tissue had no effect on body composition, weight gain, or glucose and insulin tolerance in mice fed a standard diet or a high-fat diet. In contrast, inhibition of myostatin signaling in skeletal muscle, like Mstn deletion, resulted in increased lean mass, decreased fat mass, improved glucose metabolism on standard and high-fat diets, and resistance to diet-induced obesity. Our results demonstrate that Mstn(-/-) mice have an increase in insulin sensitivity and glucose uptake, and that the reduction in adipose tissue mass in Mstn(-/-) mice is an indirect result of metabolic changes in skeletal muscle. These data suggest that increasing muscle mass by administration of myostatin antagonists may be a promising therapeutic target for treating patients with obesity or diabetes. PMID- 19295912 TI - Transcriptome analysis of synaptoneurosomes identifies neuroplasticity genes overexpressed in incipient Alzheimer's disease. AB - In Alzheimer's disease (AD), early deficits in learning and memory are a consequence of synaptic modification induced by toxic beta-amyloid oligomers (oAbeta). To identify immediate molecular targets downstream of oAbeta binding, we prepared synaptoneurosomes from prefrontal cortex of control and incipient AD (IAD) patients, and isolated mRNAs for comparison of gene expression. This novel approach concentrates synaptic mRNA, thereby increasing the ratio of synaptic to somal mRNA and allowing discrimination of expression changes in synaptically localized genes. In IAD patients, global measures of cognition declined with increasing levels of dimeric Abeta (dAbeta). These patients also showed increased expression of neuroplasticity related genes, many encoding 3'UTR consensus sequences that regulate translation in the synapse. An increase in mRNA encoding the GluR2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPAR) was paralleled by elevated expression of the corresponding protein in IAD. These results imply a functional impact on synaptic transmission as GluR2, if inserted, maintains the receptors in a low conductance state. Some overexpressed genes may induce early deficits in cognition and others compensatory mechanisms, providing targets for intervention to moderate the response to dAbeta. PMID- 19295914 TI - Impact of anti-inflammatory agents on the gene expression profile of stimulated human neutrophils: unraveling endogenous resolution pathways. AB - Adenosine, prostaglandin E(2), or increased intracellular cyclic AMP concentration each elicit potent anti-inflammatory events in human neutrophils by inhibiting functions such as phagocytosis, superoxide production, adhesion and cytokine release. However, the endogenous molecular pathways mediating these actions are poorly understood. In the present study, we examined their impact on the gene expression profile of stimulated neutrophils. Purified blood neutrophils from healthy donors were stimulated with a cocktail of inflammatory agonists in the presence of at least one of the following anti-inflammatory agents: adenosine A(2A) receptor agonist CGS 21680, prostaglandin E(2), cyclic-AMP-elevating compounds forskolin and RO 20-1724. Total RNA was analyzed using gene chips and real-time PCR. Genes encoding transcription factors, enzymes and regulatory proteins, as well as secreted cytokines/chemokines showed differential expression. We identified 15 genes for which the anti-inflammatory agents altered mRNA levels. The agents affected the expression profile in remarkably similar fashion, suggesting a central mechanism limiting cell activation. We have identified a set of genes that may be part of important resolution pathways that interfere with cell activation. Identification of these pathways will improve understanding of the capacity of tissues to terminate inflammatory responses and contribute to the development of therapeutic strategies based on endogenous resolution. PMID- 19295915 TI - A p53-dependent response limits epidermal stem cell functionality and organismal size in mice with short telomeres. AB - Telomere maintenance is essential to ensure proper size and function of organs with a high turnover. In particular, a dwarf phenotype as well as phenotypes associated to premature loss of tissue regeneration, including the skin (hair loss, hair graying, decreased wound healing), are found in mice deficient for telomerase, the enzyme responsible for maintaining telomere length. Coincidental with the appearance of these phenotypes, p53 is found activated in several tissues from these mice, where is thought to trigger cellular senescence and/or apoptotic responses. Here, we show that p53 abrogation rescues both the small size phenotype and restitutes the functionality of epidermal stem cells (ESC) of telomerase-deficient mice with dysfunctional telomeres. In particular, p53 ablation restores hair growth, skin renewal and wound healing responses upon mitogenic induction, as well as rescues ESCmobilization defects in vivo and defective ESC clonogenic activity in vitro. This recovery of ESC functions is accompanied by a downregulation of senescence markers and an increased proliferation in the skin and kidney of telomerase-deficient mice with critically short telomeres without changes in apoptosis rates. Together, these findings indicate the existence of a p53-dependent senescence response acting on stem/progenitor cells with dysfunctional telomeres that is actively limiting their contribution to tissue regeneration, thereby impinging on tissue fitness. PMID- 19295916 TI - Reverse engineering time discrete finite dynamical systems: a feasible undertaking? AB - With the advent of high-throughput profiling methods, interest in reverse engineering the structure and dynamics of biochemical networks is high. Recently an algorithm for reverse engineering of biochemical networks was developed by Laubenbacher and Stigler. It is a top-down approach using time discrete dynamical systems. One of its key steps includes the choice of a term order, a technicality imposed by the use of Grobner-bases calculations. The aim of this paper is to identify minimal requirements on data sets to be used with this algorithm and to characterize optimal data sets. We found minimal requirements on a data set based on how many terms the functions to be reverse engineered display. Furthermore, we identified optimal data sets, which we characterized using a geometric property called "general position". Moreover, we developed a constructive method to generate optimal data sets, provided a codimensional condition is fulfilled. In addition, we present a generalization of their algorithm that does not depend on the choice of a term order. For this method we derived a formula for the probability of finding the correct model, provided the data set used is optimal. We analyzed the asymptotic behavior of the probability formula for a growing number of variables n (i.e. interacting chemicals). Unfortunately, this formula converges to zero as fast as , where and . Therefore, even if an optimal data set is used and the restrictions in using term orders are overcome, the reverse engineering problem remains unfeasible, unless prodigious amounts of data are available. Such large data sets are experimentally impossible to generate with today's technologies. PMID- 19295917 TI - Dendritic cell-mediated-immunization with xenogenic PrP and adenoviral vectors breaks tolerance and prolongs mice survival against experimental scrapie. AB - In prion diseases, PrP(c), a widely expressed protein, is transformed into a pathogenic form called PrP(Sc), which is in itself infectious. Antibodies directed against PrP(c) have been shown to inhibit PrP(c) to PrP(Sc) conversion in vitro and protect in vivo from disease. Other effectors with potential to eliminate PrPSc-producing cells are cytotoxic T cells directed against PrP derived peptides but their ability to protect or to induce deleterious autoimmune reactions is not known. The natural tolerance to PrP(c) makes difficult to raise efficient adaptive responses. To break tolerance, adenovirus (Ad) encoding human PrP (hPrP) or control Ad were administered to wild-type mice by direct injection or by transfer of Ad-transduced dendritic cells (DCs). Control Ad-transduced DCs from Tg650 mice overexpressing hPrP were also used for immunization. DC-mediated but not direct administration of AdhPrP elicited antibodies that bound to murine native PrP(c). Frequencies of PrP-specific IFNgamma-secreting T cells were low and in vivo lytic activity only targeted cells strongly expressing hPrP. Immunohistochemical analysis revealed that CD3(+) T cell infiltration was similar in the brain of vaccinated and unvaccinated 139A-infected mice suggesting the absence of autoimmune reactions. Early splenic PrP(Sc) replication was strongly inhibited ten weeks post infection and mean survival time prolonged from 209 days in untreated 139A-infected mice to 246 days in mice vaccinated with DCs expressing the hPrP. The efficacy appeared to be associated with antibody but not with cytotoxic cell-mediated PrP-specific responses. PMID- 19295918 TI - Differentiation of bone marrow-derived endothelial progenitor cells is shifted into a proinflammatory phenotype by hyperglycemia. AB - Bone marrow (BM)-derived endothelial progenitor cells (EPC) contribute to vascular maintenance by participating in angiogenesis, re-endothelialization, and remodeling. Myeloid progenitor cells in the BM are functionally and quantitatively an important precursor pool for cells that contribute to these processes. However, these precursor pools in the BM also give rise to important effector cells of the innate immune system, such as macrophages and dendritic cells. We hypothesized that the disturbed repair responses that are being observed in diabetes mellitus are also related to an effect on functional and differentiation characteristics at the level of this bone marrow precursor pool. Indeed, we observed that bone marrow differentiation cultures for EPC, macrophages (Mph), or dendritic cells (DC) from hyperglycemic BM yielded 40% fewer EPC and 50% more Mph compared with control BM. These changes were directly related to the hemoglobin A(1C) levels of the donor mice. BM-derived DC numbers were not affected by hyperglycemia. The composition of the BM was not altered; in particular, the numbers of CD31+/Ly6C+ cells, which serve as common progenitors for EPC, Mph, and DC, were unaffected. In addition, BM-derived EPC from hyperglycemic mice were less angiogenic and more proinflammatory in regards to endocytosis, T-cell activation, and interleukin 12 production. HMG-CoA (3-hydroxy 3-methylglutaryl coenzyme A) reductase inhibition by statin supplementation of the culture medium counteracted these hyperglycemia-induced changes. Our study results show that hyperglycemia alters the differentiation fate of BM precursor cells, reducing the potential to generate vascular regenerative cells and favoring the development of proinflammatory cells. PMID- 19295920 TI - IDentification and Analysis of the Arabidopsis Thaliana Atfas4 Gene Whose Overexpression Results in the Development of A Fasciated Stem. AB - Using a new pEnLox vector employed to generate gain-of-function mutants in Arabidopsis thaliana, the AtFAS4 mutant has been obtained and analyzed. The mutant is characterized by super-expression of the At1g33390 gene, which leads to the occurrence of a mutant phenotype - stem fasciation. The level of expression of the AtFAS4 gene in normally developing A.thaliana plants is extremely low thus accounting for almost complete absence of information on EST's of this gene. The generated AtFAS4 mutant has permitted full-length cDNA of the At1g33390 gene to be obtained and analyzed for the first time. PMID- 19295919 TI - IGF-1 expression in infarcted myocardium and MGF E peptide actions in rat cardiomyocytes in vitro. AB - Insulinlike growth factor-1 (IGF-1) expression is implicated in myocardial pathophysiology, and two IGF-1 mRNA splice variants have been detected in rodents, IGF-1Ea and mechano-growth factor (MGF). We investigated the expression pattern of IGF-1 gene transcripts in rat myocardium from 1 h up to 8 wks after myocardial infarction induced by left anterior descending coronary artery ligation. In addition, we characterized IGF-1 and MGF E peptide action and their respective signaling in H9C2 myocardial-like cells in vitro. IGF-1Ea and MGF expression were significantly increased, both at transcriptional and translational levels, during the late postinfarction period (4 and 8 wks) in infarcted rat myocardium. Measurements of serum IGF-1 levels in infarcted rats were initially decreased (24 h up to 1 wk) but remained unaltered throughout the late experimental phase (4 to 8 wks) compared with sham-operated rats. Furthermore, specific anti-IGF-1R neutralizing antibody failed to block the synthetic MGF E peptide action, whereas it completely blocked IGF-1 action on the proliferation of H9C2 cells. Moreover, this synthetic MGF E peptide did not activate Akt phosphorylation, whereas it activated ERK1/2 in H9C2 rat myocardial cells. These data support the role of IGF-1 expression in the myocardial repair process and suggest that synthetic MGF E peptide actions may be mediated via an IGF-1R independent pathway in rat myocardial cells, as suggested by our in vitro experiments. PMID- 19295921 TI - Paradox of vaccination: is vaccination really effective against avian flu epidemics? AB - BACKGROUND: Although vaccination can be a useful tool for control of avian influenza epidemics, it might engender emergence of a vaccine-resistant strain. Field and experimental studies show that some avian influenza strains acquire resistance ability against vaccination. We investigated, in the context of the emergence of a vaccine-resistant strain, whether a vaccination program can prevent the spread of infectious disease. We also investigated how losses from immunization by vaccination imposed by the resistant strain affect the spread of the disease. METHODS AND FINDINGS: We designed and analyzed a deterministic compartment model illustrating transmission of vaccine-sensitive and vaccine resistant strains during a vaccination program. We investigated how the loss of protection effectiveness impacts the program. Results show that a vaccination to prevent the spread of disease can instead spread the disease when the resistant strain is less virulent than the sensitive strain. If the loss is high, the program does not prevent the spread of the resistant strain despite a large prevalence rate of the program. The epidemic's final size can be larger than that before the vaccination program. We propose how to use poor vaccines, which have a large loss, to maximize program effects and describe various program risks, which can be estimated using available epidemiological data. CONCLUSIONS: We presented clear and simple concepts to elucidate vaccination program guidelines to avoid negative program effects. Using our theory, monitoring the virulence of the resistant strain and investigating the loss caused by the resistant strain better development of vaccination strategies is possible. PMID- 19295923 TI - The doctor-patient relationship III: a way of listening--the Balint group revisited, 2008. PMID- 19295922 TI - Medical alert bracelet: an effective way to preserve veins for future dialysis vascular access in patients with chronic kidney disease. PMID- 19295924 TI - Sleep apnea as a cause of gout flares. PMID- 19295925 TI - Born at the wrong time? Timing of infant birth and the risk for childhood asthma. PMID- 19295926 TI - A systematic review of Cochrane anticoagulation reviews. AB - CONTEXT: I coauthored a published review of anticoagulation for venous thromboembolism in the Cochrane Database of Systematic Reviews and published a review on the same topic in MedGenMed (now the Medscape Journal of Medicine). In contrast to the article in Medscape, the discussion and conclusions in the Cochrane review were altered appreciably during the review process. Consequently, I decided to critique all anticoagulation drug-related reviews and protocols in the Cochrane database with feedback letters concerning any issues of potential controversy. EVIDENCE ACQUISITION: Using key words in the search engine of the Cochrane Reviews, I located reviews and protocols involving anticoagulant drugs. I critiqued each anticoagulation review and protocol and sent a total of 57 feedback letters to Cochrane concerning each publication to elicit a response/rebuttal from the authors. EVIDENCE SYNTHESIS: Cochrane anticoagulation review editors acknowledged receipt of all letters. As of 12 months after receipt of my last letter, the Cochrane authors have replied to 13 of the 57 and agreed with many of my points. Two protocols were withdrawn after my feedback letters were acknowledged. The 58 Cochrane anticoagulation drug reviews, including mine, contained 9 categories of methodological errors (207 total instances) and 4 types of biases (18 total instances). This review of those Cochrane reviews suggests that the effectiveness of anticoagulants for 30 medical indications is questionable. CONCLUSIONS: The efficacy of anticoagulants for treatment and prophylaxis for 30 current medical indications should be reconsidered by the scientific community and medical regulatory agencies. At least 50,000 people per year worldwide have fatal bleeding due to anticoagulant treatment or prophylaxis for these indications. PMID- 19295927 TI - Clear cell adenocarcinoma of the ovary associated with in utero diethylstilbestrol exposure: case report and clinical overview. AB - BACKGROUND: Clear cell adenocarcinoma of the vagina and cervix were previously shown to be tumors occurring in female offspring exposed prenatally to diethylstilbestrol. This report describes the first clinical case of clear cell adenocarcinoma of the ovary linked to early diethylstilbestrol exposure in utero. CASE: A 45-year-old woman presented with a self-discovered lump in the lower abdominal quadrant. She underwent surgery and staging that revealed clear cell adenocarcinoma confined to the left ovary. Foci of high-grade squamous neoplastic proliferation, inflammation, and a paratubal cyst were also present on the pathology specimen. Medical records established unequivocally that the patient's mother received diethylstilbestrol therapy throughout the pregnancy. CONCLUSION: Our case is consistent with clear cell adenocarcinoma, probably related to diethylstilbestrol exposure in utero. It reinforces the need for continued vigilance in individuals prenatally exposed to this drug. PMID- 19295928 TI - A reader responds to "Passive dissemination of printed educational materials in medicine has no or negligible effect on patient outcomes". PMID- 19295929 TI - Skin-to-skin contact provides analgesia to neonates. PMID- 19295930 TI - Part 2: a reader responds to "Does primary care matter?". PMID- 19295931 TI - Open-access medical knowledge: where are we currently and where should we be going? PMID- 19295932 TI - Let's teach moral reasoning to the next generation of doctors. PMID- 19295933 TI - Preventing nonvertebral osteoporotic fractures with extended-interval bisphosphonates: regimen selection and clinical application. AB - CONTEXT: Nonvertebral fractures (NVFs) are the most costly and disabling type of osteoporotic fractures. Bisphosphonate therapy effectively reduces the risk for NVFs; however, fracture protection depends critically on adherence and persistence. Approved bisphosphonate regimens with extended dosing intervals increase patient convenience, help patients remain on therapy, and improve fracture protection in clinical practice. EVIDENCE ACQUISITION: To assess evidence for NVF reduction with extended-interval bisphosphonates, we searched PubMed for phase 3 clinical trials, meta-analyses, and reviews of approved nitrogen-containing bisphosphonate regimens with monthly or less frequent dosing (monthly oral ibandronate, monthly or intermittent oral risedronate, quarterly intravenous [IV] ibandronate, and yearly IV zoledronic acid). These references were augmented by ISI Web of Science cited reference searches, ISI Proceedings searches, and hand searches of relevant conference proceedings and review bibliographies. EVIDENCE SYNTHESIS: Monthly oral and quarterly IV ibandronate reduce NVF risk significantly more than daily oral ibandronate and placebo, as shown by meta-analyses stratified by ibandronate dose (annual cumulative exposure). Intermittent and monthly oral risedronate have shown bone density gains similar to those seen with daily oral risedronate. Incidence rates of NVF, reported as adverse events, were also similar. Yearly IV zoledronic acid reduced NVF risk by 25% and hip fracture risk by 41% compared with placebo in its pivotal trial for postmenopausal osteoporosis. CONCLUSIONS: Extended-interval bisphosphonates offer similar or superior NVF protection with less lifestyle disruption compared with daily or weekly treatment. By removing obstacles to adherence and persistence, extended-interval oral and IV bisphosphonate regimens provide valuable therapeutic options to enhance real-world effectiveness and reduce NVF incidence. PMID- 19295934 TI - Cases in pediatric gastroenterology from The Children's Hospital of Philadelphia: a 2-year-old boy with diarrhea, failure to thrive, and hepatomegaly. PMID- 19295935 TI - Arigato, postdocs. PMID- 19295936 TI - Food availability, neighborhood socioeconomic status, and dietary patterns among blacks with type 2 diabetes mellitus. AB - CONTEXT: High diabetes prevalence among low-income and urban African American populations. OBJECTIVES & MAIN OUTCOME MEASURES: This study aimed to determine associations between neighborhood-level food sources and socioeconomic status (SES), and dietary patterns and body-mass index (BMI). The hypotheses were that the presence of food stores in neighborhoods would be associated with better dietary habits and BMI, and that the presence of convenience stores, and lower neighborhood SES, would be associated with poorer dietary habits and BMI. DESIGN, SETTING, & PATIENTS: Black adults (n = 132) with type 2 diabetes in Project Sugar 2 (Baltimore, Maryland) underwent the Ammerman dietary assessment: total dietary risk score and subscores for meat, dairy, starches, and added fat. Food source availability (food stores, convenience stores, other food stores, restaurants, and other food service places) and SES data from the 2000 US census at the tract level were linked to individual-level data. Linear mixed-effects regression models with random intercepts were used to account for neighborhood clustering and for individual-level SES and potential confounders. RESULTS: The presence of restaurants and other food service places in census tracts were associated with better dietary patterns (adjusted added fat subscore beta = -1.1, 95% confidence interval [CI] = -1.8, -0.4, and beta = -1.0, 95% CI = -1.7, -0.3, respectively). The presence of convenience stores and lower neighborhood SES was not significantly associated with worse dietary patterns or body-mass index, although trends were in the hypothesized direction. CONCLUSIONS: These findings provide some evidence for structural improvements to food environments in urban and low income black neighborhoods. PMID- 19295937 TI - Seven habits (to break) of highly effective people. PMID- 19295938 TI - A reader and author respond to "The end of peer review and traditional publishing as we know it". PMID- 19295939 TI - A primary care perspective on keloids. AB - Keloids are a common presenting complaint in the primary care clinic. This condition presents a formidable challenge, as recurrence is often difficult to prevent despite use of multiple therapeutic interventions. Part of the reason for the absence of a definitive treatment is the incomplete understanding of the pathogenesis of keloid formation, which creates a frustrating situation for both physician and patient. Here we review the most recent literature on the clinical features, pathogenesis, and management of keloids, with special emphasis on the unique challenges faced by primary care physicians. PMID- 19295940 TI - An unmet cancer screening need of HIV-positive women in southeastern Nigeria. AB - CONTEXT: HIV infection and cervical cancer are common in Nigeria. HIV-positive women have a higher risk for cervical neoplasm; therefore, counseling and regular cervical cancer screening are recommended. After post-HIV testing counseling, HIV positive women should be aware of the Papanicolaou (Pap) smear. OBJECTIVE: To determine the coverage of cervical cancer screening information for HIV-positive women by the post-HIV testing counseling. METHODS: Questionnaires were administered to 150 HIV-positive women and 150 HIV-negative controls after post HIV testing counseling at the voluntary counseling and testing clinic of University of Nigeria Teaching Hospital, Enugu, Nigeria. Proportions of responses were compared by using a chi-square test at a 95% confidence level. RESULTS: Thirty-three (22.0%) HIV-positive women and 57 (38%) HIV-negative women were aware of cervical cancer. The awareness of the Pap smear among HIV-positive women (4.0%) was lower than that of HIV-negative women (21.3%) (P < .001). However, this finding became insignificant after adjustment for educational status groups (P > .05). No respondent agreed to being informed of Pap smears during post-HIV testing counseling. After counseling on cervical cancer, 96.0% of the HIV positive group and 98.7% of the HIV-negative group expressed willingness to undergo routine Pap screening. CONCLUSIONS: Cervical cancer screening information is not part of post-HIV testing counseling for women in Enugu, southeastern Nigeria. PMID- 19295942 TI - Spontaneous recovery of adrenal insufficiency in POEMS syndrome. AB - POEMS syndrome is a rare multisystem disorder characterized by polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes. We report the first case of spontaneous recovery of adrenal insufficiency in a patient with POEMS syndrome treated with autologous hematopoietic cell transplantation. We conclude that in patients with POEMS syndrome, adrenal insufficiency should be regularly assessed for potential recovery. PMID- 19295941 TI - Differential assessment and management of asthma vs chronic obstructive pulmonary disease. AB - CONTEXT: Asthma and chronic obstructive pulmonary disease (COPD) are prevalent respiratory conditions with overlapping disease characteristics. Differentiation between asthma and COPD is important because several aspects of the guideline recommended management strategies differ for these conditions. This review identifies the epidemiologic, etiologic, and clinical distinctions of these diseases to assist physicians and other clinicians in differentiating between asthma and COPD. Key components of the guideline-recommended management approaches for these conditions are also reviewed. SEARCH STRATEGIES: Relevant articles were found by searching the MEDLINE database for "asthma" and "chronic obstructive pulmonary disease OR COPD" in association with the terms "diagnostic criteria" and "differential diagnosis". Recent statistical summaries (meta analyses), reviews, and consensus-type documents were also included. SYNTHESIS: A review of relevant articles found that, although asthma and COPD may occur simultaneously, differences between these diseases are frequently recognized in terms of age at onset, prevalence in relation to age and sex, potential for reversibility of airway obstruction, pathophysiology, and typical symptom presentation. A thorough clinical history in conjunction with lung function testing usually aids in diagnostic distinction and choice of therapeutic interventions. Radiologic imaging and inflammatory marker testing may also aid in the differentiation of these conditions. Over time, disease progression often differs between asthma and COPD. CONCLUSIONS: Although overlaps exist in the disease characteristics of asthma and COPD, careful history, physical examination, and lung function testing often reveal information that facilitates distinction between these diseases, allowing physicians and other clinicians to better tailor their therapy. PMID- 19295943 TI - Global maternal mortality: an unspeakable yet avoidable human tragedy. PMID- 19295944 TI - Low-grade endometrial stromal sarcoma with intravenous extension to the heart. AB - Endometrial stromal sarcoma (ESS) is a rare neoplasm of uterine origin. Intracardiac metastasis from this tumor is extremely infrequent. This report describes a 24-year-old woman from Yemen who had irregular vaginal bleeding shortly after spontaneous abortion. She developed left-lower-limb swelling, diagnosed by duplex scanning and magnetic resonance imaging as deep venous thrombosis in the inferior vena cava (IVC) that extended into the iliac veins on both sides, as well as the left femoral vein and right atrium. She developed acute respiratory distress, from which she recovered after transfer to the intensive care unit. Transesophageal echocardiography showed a large mass occupying the right atrium and ventricle and another mass in the right ventricular outflow tract with areas of cavitations. The tumor appeared to come from the IVC and extended through the right atrium and right ventricle into the pulmonary artery, ending in several digit-like projections. After surgical resection of the intracardiac mass, pathologic examination revealed a low-grade ESS that was confirmed by immunohistochemistry. The patient underwent panhysterectomy and IVC debridement. Pathologic examination revealed infiltrating low-grade endometrial sarcoma invading the myometrium and left adnexa, with intravenous extension into the pelvic veins and the IVC to the right side of the heart. This case shows that despite its well-known good prognostic nature, low grade ESS may behave as an aggressive malignancy. PMID- 19295945 TI - Public health and the menace of markets. PMID- 19295946 TI - Plant foods in the American diet? As we sow... PMID- 19295947 TI - Fruit and vegetable intake among adolescents and adults in the United States: percentage meeting individualized recommendations. AB - CONTEXT: Fruit and vegetable intake is an important part of a healthy diet and is associated with numerous positive health outcomes. MyPyramid provides recommendations for fruit and vegetable consumption based on individual calorie requirements as determined by an individual's age, sex, and physical activity level. OBJECTIVES: To determine (1) median fruit and vegetable consumption from all dietary sources among adolescent and adult consumers and the percentage of adolescents and adults meeting individual recommended intake levels based on caloric requirements and (2) consumption levels among various demographic groups, intake levels from subtypes of fruits and vegetables, and primary contributors to fruit and vegetable intake. DESIGN: Analysis of 2-day, 24-hour recall data from the 2003-2004 National Health and Nutrition Examination Survey (NHANES), a continuous, nationally representative, cross-sectional survey. RESULTS: This study included dietary contributions of fruits and vegetables from all dietary sources. Fewer than 1 in 10 Americans meet their calorie-specific MyPyramid fruit or vegetable recommendations. Higher intake was not observed in subgroups with higher recommendations for fruit and vegetable consumption based on caloric requirements. The primary contributors to total fruit intake were whole fruits among adults and fruit juices among adolescents. The largest single contributor to overall fruit intake was orange juice. Potatoes dominated vegetable consumption, particularly among adolescents, in whom fried potatoes increased the median vegetable intake from 0.72 cup to 1.21 cups per day. Dark green and orange vegetables and legumes accounted for a small portion of vegetable intake, and few people met the recommendations. CONCLUSIONS: Few American adolescents or adults reported consuming the recommended amounts of fruits or vegetables. Increasing consumption will probably require multifaceted approaches that augment educational campaigns with policy and environmental strategies aimed at the food system at large, from farm to plate, including schools, worksites, and retail establishments. Increasing America's fruit and vegetable consumption is an important public health strategy for weight management and reduction of risk for chronic disease. PMID- 19295948 TI - The A, B, C's of factitious disorder: a response to Turner. AB - The fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-V) is being prepared, but little attention has been accorded the category of factitious disorder, despite its presence in the manual for almost 30 years. Among relevant articles that have appeared, Turner's publication advocates retention of the category, but with new criteria. In the current paper, we reject Turner's reformulation but use the identified diagnostic dilemmas to illuminate the phenomenology of factitious disorder. We also offer a reconceptualization of the diagnosis that should better inform the preparations for DSM-V. PMID- 19295949 TI - Mesenchymal stem cells in regenerative medicine: of hopes and challenges. PMID- 19295950 TI - Surgical audit of inadvertent parathyroidectomy during total thyroidectomy: incidence, risk factors, and outcome. PMID- 19295951 TI - Trends in levonorgestrel emergency contraception use, births, and abortions: the Utah experience. AB - CONTEXT: Published reports to date have failed to demonstrate a decrease in abortion rates with increased dispersal of levonorgestrel emergency contraception (LNG EC). OBJECTIVE: To evaluate whether there is an association between statewide increases in LNG EC use and birth, fertility, and abortion rates. DESIGN: Ecological study. The number of LNG EC doses dispensed at all Planned Parenthood Association of Utah (PPAU) sites (n = 6) were obtained for 2000-2006. For this time period, birth and abortion data were obtained from the Utah Department of Health. SETTING: State of Utah. PATIENTS: Women of childbearing age. MAIN OUTCOME MEASURES: Birth rates were calculated as the number of live births per 1000 population; general fertility rates, abortion rates, and LNG EC rates were calculated per 1000 women of childbearing age (15-44 years). RESULTS: Between 2000 and 2006, yearly distribution of LNG EC increased from 11,263 to 52,083 doses. Over this period, the rate of Plan B use per 1000 women age 15-44 years increased from 21.30 doses/1000 to 87.82 doses/1000, an increase of 312%. During the same period, there were corresponding changes in the statewide birth rate (-2.94%), general fertility rate (0.73%), and abortion rate (-6.36%). Pearson correlation coefficients were statistically significant for the association between the LNG EC rate and the birth rate (-0.9053; P = .0050) and the abortion rate (-0.8749; P < .001), but not between the Plan B rate and the general fertility rate (0.2446; P = .5970). CONCLUSION: This ecological study represents, to the authors' knowledge, the first statistically significant association between increasing rates of LNG EC distribution and decreasing abortion rates. PMID- 19295952 TI - A reader's response to "Mustard gas poisoning of hundreds of Americans in the United States in 1996". PMID- 19295953 TI - A diabetes drama for President Obama. PMID- 19295954 TI - So long but not farewell: the Medscape Journal of Medicine (1999-2009). PMID- 19295955 TI - Sodium replacement and plasma sodium drop during exercise in the heat when fluid intake matches fluid loss. AB - CONTEXT: Sodium replacement during prolonged exercise in the heat may be critically important to maintaining fluid and electrolyte balance and muscle contractility. OBJECTIVE: To examine the effectiveness of sodium-containing sports drinks in preventing hyponatremia and muscle cramping during prolonged exercise in the heat. DESIGN: Randomized crossover study. PATIENTS OR OTHER PARTICIPANTS: Thirteen active men. INTERVENTION(S): Participants completed 4 trials of an exercise protocol in the heat (30 degrees C) consisting of 3 hours of exercise (alternating 30 minutes of walking and cycling at a heart rate of 130 and 140 beats per minute, respectively); a set of standing calf raises (8 sets of 30 repetitions); and 45 minutes of steep, brisk walking (5.5 km x h(-1) on a 12% grade). During exercise, participants consumed fluids to match body mass loss. A different drink was consumed for each trial: carbohydrate-electrolyte drink containing 36.2 mmol/L sodium (HNa), carbohydrate-electrolyte drink containing 19.9 mmol/L sodium (LNa), mineral water (W), and colored and flavored distilled water (PL). MAIN OUTCOME MEASURE(S): Serum sodium, plasma osmolality, plasma volume changes, and muscle cramping frequency. RESULTS: During both HNa and LNa trials, serum sodium remained relatively constant (serum sodium concentration at the end of the protocol was 137.3 mmol/L and 136.7 mmol/L, respectively). However, a clear decrease was observed in W (134.5 +/- 0.8 mmol/L) and PL (134.4 +/- 0.8 mmol/L) trials compared with HNa and LNa trials (P < .05). The same trends were observed for plasma osmolality (P < .05). Albeit not significant, plasma volume was preserved during the HNa and LNa trials, but a reduction of 2.5% was observed in the W and PL trials. None of the volunteers experienced cramping. CONCLUSIONS: The data suggest that sodium intake during prolonged exercise in the heat plays a significant role in preventing sodium losses that may lead to hyponatremia when fluid intake matches sweat losses. PMID- 19295956 TI - Validity and reliability of devices that assess body temperature during indoor exercise in the heat. AB - CONTEXT: When assessing exercise hyperthermia outdoors, the validity of certain commonly used body temperature measuring devices has been questioned. A controlled laboratory environment is generally less influenced by environmental factors (eg, ambient temperature, solar radiation, wind) than an outdoor setting. The validity of these temperature measuring devices in a controlled environment may be more acceptable. OBJECTIVE: To assess the validity and reliability of commonly used temperature devices compared with rectal temperature in individuals exercising in a controlled, high environmental temperature indoor setting and then resting in a cool environment. DESIGN: Time series study. SETTING: Laboratory environmental chamber (temperature = 36.4 +/- 1.2 degrees C [97.5 +/- 2.16 degrees F], relative humidity = 52%) and cool laboratory (temperature = approximately 23.3 degrees C [74.0 degrees F], relative humidity = 40%). PATIENTS OR OTHER PARTICIPANTS: Fifteen males and 10 females. INTERVENTION(S): Rectal, gastrointestinal, forehead, oral, aural, temporal, and axillary temperatures were measured with commonly used temperature devices. Temperature was measured before and 20 minutes after entering the environmental chamber, every 30 minutes during a 90-minute treadmill walk in the heat, and every 20 minutes during a 60-minute rest in mild conditions. Device validity and reliability were assessed with various statistical measures to compare the measurements using each device with rectal temperature. A device was considered invalid if the mean bias (average difference between rectal and device temperatures) was more than +/-0.27 degrees C (+/-0.50 degrees F). MAIN OUTCOME MEASURE(S): Measured temperature from each device (mean and across time). RESULTS: The following devices provided invalid estimates of rectal temperature: forehead sticker (0.29 degrees C [0.52 degrees F]), oral temperature using an inexpensive device (-1.13 degrees C [-2.03 degrees F]), temporal temperature measured according to the instruction manual (-0.87 degrees C [-1.56 degrees F]), temporal temperature using a modified technique ( 0.63 degrees C [-1.13 degrees F]), oral temperature using an expensive device ( 0.86 degrees C, [-1.55 degrees F]), aural temperature (-0.67 degrees C, [-1.20 degrees F]), axillary temperature using an inexpensive device (-1.25 degrees C, [ 2.24 degrees F]), and axillary temperature using an expensive device (-0.94 degrees F [-1.70 degrees F]). Measurement of intestinal temperature (mean bias of -0.02 degrees C [-0.03 degrees F]) was the only device considered valid. Devices measured in succession (intestinal, forehead, temporal, and aural) showed acceptable reliability (all had a mean bias = 0.09 degrees C [0.16 degrees F] and r >or= 0.94]). CONCLUSIONS: Even during laboratory exercise in a controlled environment, devices used to measure forehead, temporal, oral, aural, and axillary body sites did not provide valid estimates of rectal temperature. Only intestinal temperature measurement met the criterion. Therefore, we recommend that rectal or intestinal temperature be used to assess hyperthermia in individuals exercising indoors in the heat. PMID- 19295957 TI - Comparisons of cubed ice, crushed ice, and wetted ice on intramuscular and surface temperature changes. AB - CONTEXT: Many researchers have investigated the effectiveness of different types of cold application, including cold whirlpools, ice packs, and chemical packs. However, few have investigated the effectiveness of different types of ice used in ice packs, even though ice is one of the most common forms of cold application. OBJECTIVE: To evaluate and compare the cooling effectiveness of ice packs made with cubed, crushed, and wetted ice on intramuscular and skin surface temperatures. DESIGN: Repeated-measures counterbalanced design. SETTING: Human performance research laboratory. PATIENTS OR OTHER PARTICIPANTS: Twelve healthy participants (6 men, 6 women) with no history of musculoskeletal disease and no known preexisting inflammatory conditions or recent orthopaedic injuries to the lower extremities. INTERVENTION(S): Ice packs made with cubed, crushed, or wetted ice were applied to a standardized area on the posterior aspect of the right gastrocnemius for 20 minutes. Each participant was given separate ice pack treatments, with at least 4 days between treatment sessions. MAIN OUTCOME MEASURE(S): Cutaneous and intramuscular (2 cm plus one-half skinfold measurement) temperatures of the right gastrocnemius were measured every 30 seconds during a 20-minute baseline period, a 20-minute treatment period, and a 120-minute recovery period. RESULTS: Differences were observed among all treatments. Compared with the crushed-ice treatment, the cubed-ice and wetted-ice treatments produced lower surface and intramuscular temperatures. Wetted ice produced the greatest overall temperature change during treatment and recovery, and crushed ice produced the smallest change. CONCLUSIONS: As administered in our protocol, wetted ice was superior to cubed or crushed ice at reducing surface temperatures, whereas both cubed ice and wetted ice were superior to crushed ice at reducing intramuscular temperatures. PMID- 19295958 TI - Ultrasound techniques applied to body fat measurement in male and female athletes. AB - CONTEXT: For athletes in disciplines with weight categories, it is important to assess body composition and weight fluctuations. OBJECTIVE: To evaluate the accuracy of measuring body fat percentage with a portable ultrasound device possessing high accuracy and reliability versus fan-beam, dual-energy X-ray absorptiometry (DEXA). DESIGN: Cross-validation study. SETTING: Research laboratory. PATIENTS OR OTHER PARTICIPANTS: A total of 93 athletes (24 women, 69 men), aged 23.5 +/- 3.7 years, with body mass index = 24.0 +/- 4.2 and body fat percentage via DEXA = 9.41 +/- 8.1 participated. All participants were elite athletes selected from the Institut National des Sports et de l'Education Physique. These participants practiced a variety of weight-category sports. MAIN OUTCOME MEASURE(S): We measured body fat and body fat percentage using an ultrasound technique associated with anthropometric values and the DEXA reference technique. Cross-validation between the ultrasound technique and DEXA was then performed. RESULTS: Ultrasound estimates of body fat percentage were correlated closely with those of DEXA in both females (r = 0.97, standard error of the estimate = 1.79) and males (r = 0.98, standard error of the estimate = 0.96). The ultrasound technique in both sexes had a low total error (0.93). The 95% limit of agreement was -0.06 +/- 1.2 for all athletes and did not show an overprediction or underprediction bias. We developed a new model to produce body fat estimates with ultrasound and anthropometric dimensions. CONCLUSIONS: The limits of agreement with the ultrasound technique compared with DEXA measurements were very good. Consequently, the use of a portable ultrasound device produced accurate body fat and body fat percentage estimates in relation to the fan-beam DEXA technique. PMID- 19295959 TI - Rotator cuff tear pain and tear size and scapulohumeral rhythm. AB - CONTEXT: The body of knowledge concerning shoulder kinematics in patients with rotator cuff tears is increasing. However, the level of understanding regarding how pain and tear size affect these kinematic patterns is minimal. OBJECTIVE: To identify relationships between pain associated with a full-thickness rotator cuff tear, tear size, and scapulohumeral rhythm (SHR) and to determine whether pain and tear size serve as predictors of SHR. DESIGN: A test-retest design was used to quantify pain and SHR before and after a subacromial lidocaine injection. Correlation and multivariate analyses were used to identify relationships among pain, tear size, and SHR. SETTING: Orthopaedic biomechanics research laboratory. PATIENTS OR OTHER PARTICIPANTS: Fifteen patients (age range, 40-75 years) with diagnosed full-thickness rotator cuff tears participated. They were experiencing pain at the time of testing. INTERVENTION(S): Shoulder kinematic data were collected with an electromagnetic tracking system before and after the patient received a lidocaine injection. MAIN OUTCOME MEASURE(S): Pain was rated using a visual analog scale. Three-dimensional scapular kinematics and glenohumeral elevation were assessed. Scapular kinematics included anterior-posterior tilt, medial-lateral tilt, and upward-downward rotation. A regression model was used to calculate SHR (scapular kinematics to glenohumeral elevation) for phases of humeral elevation and lowering. RESULTS: Linear relationships were identified between initial pain scores and SHR and between tear size and SHR, representing an increased reliance on scapular motion with increasing pain and tear size. Pain was identified as an independent predictor of SHR, whereas significant findings for the effect of tear size on SHR and the interaction between pain and tear size were limited. CONCLUSIONS: We noted an increased reliance on scapular contributions to overall humeral elevation with increasing levels of pain and rotator cuff tear size. Pain associated with a rotator cuff tear serves as a primary contributor to the kinematic patterns exhibited in patients with rotator cuff tears. PMID- 19295960 TI - A clinical method for identifying scapular dyskinesis, part 1: reliability. AB - CONTEXT: Shoulder injuries are common in athletes involved in overhead sports, and scapular dyskinesis is believed to be one causative factor in these injuries. Many authors assert that abnormal scapular motion, so-called dyskinesis, is related to shoulder injury, but evidence from 3-dimensional measurement studies regarding this relationship is mixed. Reliable and valid clinical methods for detecting scapular dyskinesis are lacking. OBJECTIVE: To determine the interrater reliability of a new test designed to detect abnormal scapular motion. DESIGN: Correlation design using ratings from multiple pairs of testers. SETTING: University athletic training facilities. PATIENTS OR OTHER PARTICIPANTS: A sample of 142 athletes (from National Collegiate Athletic Association Divisions I and III) participating in sports requiring intense overhead arm use. INTERVENTION(S): Participants were videotaped from the posterior aspect while performing 5 repetitions of bilateral, weighted (1.4-kg [3-lb] or 2.3-kg [5-lb]) shoulder flexion and frontal-plane abduction. Videotapes from randomly chosen participants were subsequently viewed and independently rated for the presence of scapular dyskinesis by 6 raters (3 pairs), with each pair rating 30 different participants. Raters were trained to detect scapular dyskinesis using a self instructional format with standardized operational definitions and videotaped examples of normal and abnormal motion. MAIN OUTCOME MEASURE(S): Scapular dyskinesis was defined as the presence of either winging or dysrhythmia. Right and left sides were rated independently as normal, subtle, or obvious dyskinesis. We calculated percentage of agreement and weighted kappa (kappa(w)) coefficients to determine reliability. RESULTS: Percentage of agreement was between 75% and 82%, and kappa(w) ranged from 0.48 to 0.61. CONCLUSIONS: The test for scapular dyskinesis showed satisfactory reliability for clinical use in a sample of overhead athletes known to be at increased risk for shoulder symptoms. PMID- 19295961 TI - A clinical method for identifying scapular dyskinesis, part 2: validity. AB - CONTEXT: Although clinical methods for detecting scapular dyskinesis have been described, evidence supporting the validity of these methods is lacking. OBJECTIVE: To determine the validity of the scapular dyskinesis test, a visually based method of identifying abnormal scapular motion. A secondary purpose was to explore the relationship between scapular dyskinesis and shoulder symptoms. DESIGN: Validation study comparing 3-dimensional measures of scapular motion among participants clinically judged as having either normal motion or scapular dyskinesis. SETTING: University athletic training facilities. PATIENTS OR OTHER PARTICIPANTS: A sample of 142 collegiate athletes (National Collegiate Athletic Association Division I and Division III) participating in sports requiring overhead use of the arm was rated, and 66 of these underwent 3-dimensional testing. INTERVENTION(S): Volunteers were viewed by 2 raters while performing weighted shoulder flexion and abduction. The right and left sides were rated independently as normal, subtle dyskinesis, or obvious dyskinesis using the scapular dyskinesis test. Symptoms were assessed using the Penn Shoulder Score. MAIN OUTCOME MEASURE(S): Athletes judged as having either normal motion or obvious dyskinesis underwent 3-dimensional electromagnetic kinematic testing while performing the same movements. The kinematic data from both groups were compared via multifactor analysis of variance with post hoc testing using the least significant difference procedure. The relationship between symptoms and scapular dyskinesis was evaluated by odds ratios. RESULTS: Differences were found between the normal and obvious dyskinesis groups. Participants with obvious dyskinesis showed less scapular upward rotation (P < .001), less clavicular elevation (P < .001), and greater clavicular protraction (P = .044). The presence of shoulder symptoms was not different between the normal and obvious dyskinesis volunteers (odds ratio = 0.79, 95% confidence interval = 0.33, 1.89). CONCLUSIONS: Shoulders visually judged as having dyskinesis showed distinct alterations in 3-dimensional scapular motion. However, the presence of scapular dyskinesis was not related to shoulder symptoms in athletes engaged in overhead sports. PMID- 19295962 TI - Sagittal-plane trunk position, landing forces, and quadriceps electromyographic activity. AB - CONTEXT: Researchers have suggested that large landing forces, excessive quadriceps activity, and an erect posture during landing are risk factors for anterior cruciate ligament (ACL) injury. The influence of knee kinematics on these risk factors has been investigated extensively, but trunk positioning has received little attention. OBJECTIVE: To determine the effect of trunk flexion on landing forces and quadriceps activation during landing. DESIGN: Two (sex) x 2 (task) repeated-measures design. SETTING: Research laboratory. PATIENTS OR OTHER PARTICIPANTS: Forty healthy, physically active volunteers (20 men, 20 women). INTERVENTION(S): Participants performed 2 drop-landing tasks. The first task represented the natural, or preferred, landing strategy. The second task was identical to the first except that participants flexed the trunk during landing. MAIN OUTCOME MEASURE(S): We measured peak vertical and posterior ground reaction forces and mean quadriceps electromyographic amplitude during the loading phase of landing (ie, the interval from initial ground contact to peak knee flexion). RESULTS: Trunk flexion decreased the vertical ground reaction force (P < .001) and quadriceps electromyographic amplitude (P < .001). The effect of trunk flexion did not differ across sex for landing forces or quadriceps electromyographic activity. CONCLUSIONS: We found that trunk flexion during landing reduced landing forces and quadriceps activity, thus potentially reducing the force imparted to the ACL. Research has indicated that trunk flexion during landing also increases knee and hip flexion, resulting in a less erect landing posture. In combination, these findings support emphasis on trunk flexion during landing as part of ACL injury-prevention programs. PMID- 19295963 TI - Soccer-specific fatigue and eccentric hamstrings muscle strength. AB - CONTEXT: Epidemiologic findings of higher incidences of hamstrings muscle strains during the latter stages of soccer match play have been attributed to fatigue. OBJECTIVE: To investigate the influence of soccer-specific fatigue on the peak eccentric torque of the knee flexor muscles. DESIGN: Descriptive laboratory study. SETTING: Controlled laboratory environment. PATIENTS OR OTHER PARTICIPANTS: Ten male professional soccer players (age = 24.7 +/- 4.4 years, mass = 77.1 +/- 8.3 kg, Vo(2max) = 63.0 +/- 4.8 mL x kg(-1) x min(-1)). INTERVENTION(S): Participants completed an intermittent treadmill protocol replicating the activity profile of soccer match play, with a passive halftime interval. Before exercise and at 15-minute intervals, each player completed isokinetic dynamometer trials. MAIN OUTCOME MEASURE(S): Peak eccentric knee flexor torque was quantified at isokinetic speeds of 180 degrees x s(-1), 300 degrees x s(-1), and 60 degrees x s(-1), with 5 repetitions at each speed. RESULTS: Peak eccentric knee flexor torque at the end of the game (T(300eccH105) = 127 +/- 25 Nm) and at the end of the passive halftime interval (T(300eccH60) = 133 +/- 32 Nm) was reduced relative to T(300eccH00) (167 +/- 35 Nm, P < .01) and T(300eccH15) (161 +/- 35 Nm, P = .02). CONCLUSIONS: Eccentric hamstrings strength decreased as a function of time and after the halftime interval. This finding indicates a greater risk of injuries at these specific times, especially for explosive movements, in accordance with epidemiologic observations. Incorporating eccentric knee flexor exercises into resistance training sessions that follow soccer-specific conditioning is warranted to try to reduce the incidence or recurrence of hamstrings strains. PMID- 19295964 TI - Development and preliminary validation of the Injury-Psychological Readiness to Return to Sport (I-PRRS) scale. AB - CONTEXT: Only a few scales measure confidence within sport; however, these scales are insufficient to measure confidence after athletic injuries. Therefore, better measures are needed to determine the psychological readiness of injured athletes to return to sport participation. OBJECTIVE: To develop a scale that measures the psychological readiness of injured athletes to return to sport participation and to provide preliminary evidence of reliability and validity for the scale. DESIGN: The Delphi method was used to develop the Injury-Psychological Readiness to Return to Sport scale (I-PRRS). Two 1-way analyses of variance with repeated measures and 6 Pearson product moment correlations were computed to help validate the scale. SETTING: Athletic training clinics at 3 National Collegiate Athletic Association (NCAA) schools. PATIENTS OR OTHER PARTICIPANTS: Four certified athletic trainers (ATs) and professors of Commission on Accreditation of Athletic Training Education-accredited athletic training programs and 3 NCAA Division III coaches made up a panel of experts that participated in the Delphi portion of the study to develop the I-PRRS. In the second part of the study, 22 injured athletes, who missed a minimum of 1 week of practice, from 3 NCAA schools in Divisions II and III were surveyed along with their respective ATs. The injured athletes and ATs participated in the validation of the I-PRRS. MAIN OUTCOME MEASURE(S): The injured athlete completed the Profile of Mood States (POMS) short form and the I-PRRS shortly after injury, before returning to the first practice, before returning to competition, and immediately after competition. The respective AT completed the I-PRRS before and after competition. The I-PRRS is a 6-item scale that measures the psychological readiness of injured athletes to return to sport, and the POMS short form is a 30-item scale that measures mood states. I added the negative moods of the POMS and subtracted the positive moods of the POMS to calculate a Total Mood Disturbance (TMD) score. RESULTS: The I PRRS scores were negatively correlated with the TMD scores of the POMS short form at all 4 time intervals, showing concurrent validity. The I-PRRS scores were lowest after injury, increased before practice, increased again before competition, and had no change after competition. The I-PRRS as completed by the athlete and respective AT was positively correlated both before and after practice, demonstrating external validity. CONCLUSIONS: Preliminary evidence for reliability and validity of the I-PRRS was demonstrated. The I-PRRS can be a beneficial tool for ATs to assess an athlete's psychological readiness to return to sport participation after injury. PMID- 19295965 TI - An assessment of burnout in undergraduate athletic training education program directors. AB - CONTEXT: Athletic training education program directors (ATEPDs) often manage their time among students, program administration, and patient care. OBJECTIVE: To assess the level of burnout in ATEPDs and to determine the relationship between burnout and various demographics of ATEPDs. DESIGN: Cross-sectional study. SETTING: Public and private colleges and universities nationwide. PATIENTS OR OTHER PARTICIPANTS: Two hundred forty-nine ATEPDs of undergraduate athletic training education programs accredited by the Commission on Accreditation of Athletic Training Education. INTERVENTION(S): We administered the Maslach Burnout Inventory (MBI) to all participants. MAIN OUTCOME MEASURE(S): The MBI consisted of 21 items assessing 3 characteristics of burnout: emotional exhaustion, depersonalization, and personal accomplishment. Another component of the survey requested demographic information about the ATEPDs. We used univariate, multivariate, and factorial analyses of variance with the alpha level set a priori at .05. We also calculated Pearson product moment correlation coefficients. RESULTS: Women had greater emotional exhaustion than men (20.67 +/- 9.43 and 16.47 +/- 9.64, respectively) (P = .001). The difference between tenure status groups for emotional exhaustion was significant (P = .014), with tenure track ATEPDs scoring higher on emotional exhaustion than tenured ATEPDs. Pearson product moment correlation coefficients revealed a weak negative relationship among emotional exhaustion and age (r = -0.263, P < .001), years of program director experience (r = -0.157, P = .013), and years at current job (r = -0.162, P = .010), indicating that as ATEPDs aged, gained more experience, and stayed in their current jobs, their emotional exhaustion scores decreased. There was also a weak negative relationship between age and depersonalization (r = -0.171, P = .007). There was a weak positive relationship between years at current job and personal accomplishment (r = 0.197, P = .002). CONCLUSIONS: We found that ATEPDs experienced a moderate form of emotional exhaustion burnout and low depersonalization and personal accomplishment burnout, with women experiencing greater emotional exhaustion than males. Additionally, ATEPDs in tenure-track positions experienced greater emotional exhaustion than tenured ATEPDs. The ATEPDs need to obtain healthy coping strategies early within their directorships to manage components related to burnout. PMID- 19295966 TI - Student retention in athletic training education programs. AB - CONTEXT: The success of any academic program, including athletic training, depends upon attracting and keeping quality students. The nature of persistent students versus students who prematurely leave the athletic training major is not known. Understanding the profiles of athletic training students who persist or leave is important. OBJECTIVE: To (1) explore the relationships among the following variables: anticipatory factors, academic integration, clinical integration, social integration, and motivation; (2) determine which of the aforementioned variables discriminate between senior athletic training students and major changers; and (3) identify which variable is the strongest predictor of persistence in athletic training education programs. DESIGN: Descriptive study using a qualitative and quantitative mixed-methods approach. SETTING: Thirteen athletic training education programs located in District 3 of the National Athletic Trainers' Association. PATIENTS OR OTHER PARTICIPANTS: Ninety-four senior-level athletic training students and 31 college students who changed majors from athletic training to another degree option. DATA COLLECTION: Data were collected with the Athletic Training Education Program Student Retention Questionnaire (ATEPSRQ). ANALYSIS: Data from the ATEPSRQ were analyzed via Pearson correlations, multivariate analysis of variance, univariate analysis of variance, and a stepwise discriminant analysis. Open-ended questions were transcribed and analyzed using open, axial, and selective coding procedures. Member checks and peer debriefing techniques ensured trustworthiness of the study. RESULTS: Pearson correlations identified moderate relationships among motivation and clinical integration (r = 0.515, P < .01) and motivation and academic integration (r = 0.509, P < .01). Univariate analyses of variance showed that academic integration (F(1,122) = 8.483, P < .004), clinical integration (F(1,119) = 30.214, P < .001), and motivation (F(1,121) = 68.887, P < .001) discriminated between seniors and major changers. Discriminant analysis indicated that motivation was the strongest predictor of persistence in athletic training education, accounting for 37.2% of the variance between groups. The theoretic model accurately classified 95.7% of the seniors and 53.8% of the major changers. A common theme emerging from the qualitative data was the presence of a strong peer-support group that surrounded many of the senior-level students. CONCLUSIONS: Understanding student retention in athletic training is important for our profession. Results from this study suggest 3 key factors associated with student persistence in athletic training education programs: (1) student motivation, (2) clinical and academic integration, and (3) the presence of a peer support system. Educators and program directors must create comprehensive recruitment and retention strategies that address factors influencing students' decisions to stay in the athletic training profession. PMID- 19295967 TI - Ice hockey injuries in a Japanese elite team: a 3-year prospective study. AB - CONTEXT: As the Asian Ice Hockey League gradually expands and becomes more competitive, ice hockey-related injuries may increase. However, no reports have been published on ice hockey injuries in Japan, including the method of injury and the daily supervision of the players during the regular season. OBJECTIVE: To prospectively study the incidence, types, and mechanisms of ice hockey injuries in an elite Japanese ice hockey team. DESIGN: Prospective observational cohort study design. SETTING: An elite ice hockey team, Tokyo, Japan. PATIENTS OR OTHER PARTICIPANTS: Ninety-four players during the 2002-2005 seasons. MAIN OUTCOME MEASURE(S): Data were collected for 3 consecutive seasons using an injury reporting form. RESULTS: The overall game injury rate was 74.3 per 1000 player game hours and 11.7 per 1000 player-game hours for injuries resulting in any time loss. The overall practice injury rates were 11.2 per 1000 player-practice hours and 1.1 per 1000 player-practice hours for injuries resulting in any time loss. Forwards had the highest rate of injury, followed by defensemen and then goalkeepers. Contusions were the most common injury, followed by strains, lacerations, and sprains. CONCLUSIONS: Most injuries among Japanese ice hockey players occurred during games. Game or play intensity may influence the injury rate during games. PMID- 19295968 TI - Does creatine supplementation hinder exercise heat tolerance or hydration status? A systematic review with meta-analyses. AB - OBJECTIVE: To critically assess original research addressing the effect of creatine supplementation on exercise heat tolerance and hydration status. DATA SOURCES: We searched the electronic databases PubMed, Scopus, Web of Science, SPORTDiscus, and Rehabilitation & Physical Medicine, without date limitations, for the following key words: creatine, exercise, thermoregulation, dehydration, hyperthermia, heat tolerance, exertional heat illnesses, and renal function. Our goal was to identify randomized clinical trials investigating the effect of creatine supplementation on hydration status and thermoregulation. Citations from related articles also were identified and retrieved. DATA SYNTHESIS: Original research was reviewed using the Physiotherapy Evidence Database (PEDro) Scale. One author initially screened all articles. Fifteen of 95 articles examined the effects of creatine on thermoregulation or hydration status (or both). Two independent reviewers then reviewed these articles. Ten studies were selected on the basis of inclusion and exclusion criteria. The PEDro scores for the 10 studies ranged from 7 to 10 points (maximum possible score = 10 points). CONCLUSIONS: No evidence supports the concept that creatine supplementation either hinders the body's ability to dissipate heat or negatively affects the athlete's body fluid balance. Controlled experimental trials of athletes exercising in the heat resulted in no adverse effects from creatine supplementation at recommended dosages. PMID- 19295969 TI - Myocardial perfusion after one year of spinal cord stimulation in patients with refractory angina. AB - AIM: Spinal cord stimulation (SCS) is recommended for patients with coronary artery disease (CAD) and refractory angina. We used positron emission tomography (PET) to investigate the long-term effect of SCS on regional myocardial perfusion in patients suffering from angina pectoris refractory to medical treatment and without option for coronary intervention. PATIENTS, METHODS: We analyzed data of 44 patients with stable CAD (91% three vessel disease). At baseline, we determined coronary flow reserve (CFR) using 13N-ammonia-PET and myocardial viability with 18F-FDG. SCS was performed for one year (Medtronic Itrell III or Synergy, Dusseldorf, Germany). During follow-up, no cardiac interventions were necessary and no myocardial infarctions occurred. At one year follow-up, CFR was measured again. RESULTS: In the majority of patients (77%), SCS led to an improvement of clinical symptoms. CFR did not change significantly during follow up. Subjective improvement did not correlate with an increase of CFR. CONCLUSIONS: Despite its clinical effect, SCS does not have a direct impact on CFR in patients with stable CAD. According to our results, the pain relief is not due to an improvement of the myocardial blood supply. PMID- 19295970 TI - [Hospitalizations due to decompensated cardiac insufficiency in Spain]. PMID- 19295971 TI - [Epidemiological events related to decompensated heart failure]. AB - OBJECTIVE: The present study aimed to evaluate the profile of patients with decompensated heart failure hospitalized in a tertiary hospital. METHODS: It was designed an observational and retrospective study where data from clinical records of patients suffering from heart failure along 2005 were registered randomly. RESULTS: 209 patients were collected (average age: 78.6 +/- 9.1; male: 52.4%) with a comorbidity rate of 87.55%. Almost one third of them have not stimation of systolic function and among the others 72.4% have it preserved. Most of decompensated were due to respiratory infections. Ischemic-hipertensive cardiopathy was the most frequent aetiology of systolic disfunction. Average stay was 12.9 days with a mortality rate of 9.56%. Its main risk factors were advanced stages in NYHA od Red Cross scales, as so as dementia or ictus. CONCLUSIONS: The present study shows a patient hospitalized for decompensated heart failure roughly different from that one reported at clinical trials. It makes difficult to apply therapeutical interventions, previously well documented to be useful. PMID- 19295972 TI - [Epidemiologic survey of acute poisoning in the south area of the Community of Madrid: the VEIA 2004 study]. AB - OBJECTIVE: VEIA study is an evolutional registry of acute poisonings (AP) attended in the Emergency Room of the Doce de Octubre Hospital in Madrid (Spain) in a whole year (1979, 1985, 1990, 1994,1997 and 2000). We present the 2004 study and compare the results with the previous years. METHODS AND RESULTS: Methodology has been identical across VEIA STUDY. Of 1508 AP, 610 are suicide attempts (IAVIS), 319 ethylic, (IAVE), and 218 by illicit drugs (IAVD). Of the 2,259 toxics involved, 48% are medications (50% benzodiacepines) alcohol 25% and illicit drugs 13%. CONCLUSIONS: There is an important increase (34%) of cases and the incidence surpasses all published in our country. There are no gender differences nor in the whole neither the Health Area Census, but there are differences in IAVIS, IAVE and IAVD. IAVIS increase in 35%. Benzodiacepines poisoning increases two-fold as well as antidepressive drugs do in women. In men also increase, but in a minor extent. Acetaminophen remains the same in 23%. NSAID's, adjuvants and myorelaxants increase in women as do also alcohol and other poisons that almost equal men's. There are 13 cases of IAVIS in patients with alimentary disorders. Among men, a quarter are illicit drug abusers. In IAVE, the group without alcoholism grows and the total decreases. Illicit drugs duplicate the number of the former year. Cocaine supposes already 2/3 of the cases, MDMA ascends to 22 cases and they appear new substances as glue, without doubt as an effect of immigration and ketamine. Finally 205 household accidents and 57 industrial injuries complete the series. PMID- 19295973 TI - [Study of the conpsumption of innapropiate medicaments in elder hospitalized in the Internal Medicine Service]. AB - BACKGROUND: The elderly patients are characterized by the high degree of polymedication. This factor is the most important in the appearance of adverse effects (EAD). The study of the medicaments helps to establish which are inappropriate and therefore they must move back, diminishing of this form EAD's possibility. PATIENTS AND METHOD: Prospective and observacional Study. Criterion of incorporation: patients of > 64 years old, hospitalized in an Internal Medicine Service. There were defined as inappropriate medicament the medicines of low therapeutic utility, them not indicated, and the inadequate ones for the elder. By means of the SPSS 11.5 program the possible variables related with the consumption were analyzed. RESULTS: 172 elders were included in the study. The average of medicines for person and day belonged to 5.34 (0-15). 52.5% was consuming an inappropriate medicament (36.6% inadequate for the elder, 15% not indicated, and 12% UTBs). The analysis multivariant associates the consumption of these medicaments with a major number of diseases (p < 0.012), to a major consumption of medicines (p < 0.001) and to the origin of the nursing residences (p < 0.001). Only the consumption of medicaments not adapted for the elder is associated with increase of EAD. CONCLUSION: The half of the elders takes at least a medicament of unnecessary form, and the majority of these medicaments favors appearance of EAD. PMID- 19295974 TI - [Influence of a W3 fatty acids oral enhanced formula in clinical and biochemical parameters of head and neck cancer ambulatory patients]. AB - INTRODUCTION: Postsurgical patients with head and neck cancer could have a high rate of ambulatory complications. The aim of our study was to investigate whether oral ambulatory nutrition of post surgical head and neck cancer patients with recent weight loss, using an omega 3 fatty acids and arginine diet could improve nutritional status and complications. DESIGN: A population of 30 ambulatory post surgical patients with oral and laryngeal cancer and recent weight loss was enrolled. At Hospital discharge post surgical head and neck cancer patients were asked to consume two cans per day of a specially designed omega 3 fatty acid and arginine enhanced supplement during 12 weeks. RESULTS: The average age was 62.7 + 11.1 years (3 females/27 males). The average days of dietary suplementation was 90.8 + 22 days. Blood albumin, prealbumin, transferring and lymphocites concentrations improved. C reactive protein decreased. Neither weight improvement nor other anthropometric parameters improved. A positive correlation between ratio w3/w6and kg of fat mass (r = 0.69; p < 0.05) and with midarm circumference (r = 0.62; p < 0.05) were detected. Patients were divided in two groups; group I (ratio dietary intake w3/w6 < 0,51) and group II (ratio dietary intake w3/w6 > 0,51). Group II had more final fat mass than group I (17.52 + 6.3 kg vs. 13.9 + 5.1 kg; p < 0.05). CONCLUSION: At dose taken, inmuno-enhanced formula increased blood proteins (albumin, prealbumin and transferrin) concentrations and decresed CRP. The increase of fat mass was higher in patients with better w3/w6 ratios. PMID- 19295975 TI - [Visceral leishmaniasis and bronchial asthma: influence of steroid therapy in the development of the macrophage activation syndrome and relative adrenal insufficiency]. AB - The risk of suffering opportunistics infections in the immunoincompetent patient is a fact perfectly established. An uncommon situation constitutes the bronchopaties, pathologies with a high prevalence among the general population that they require habitually, among other, steroid treatment. The immunosupression confers to the clinical evolution of the infections, as a consequence of the inadequate response to the physical stress, due to the inhibition of the hypothalamus-hypophysis axis being able to in particularly serious cases, to develop the denominated macrophage activation syndrome, a serious and uncommon syndrome that darkens the clinical prognosis in these patients. In presence of a feverish syndrome of uncertain origin in a patient in immunosuppressor treatment, although it is to low dose, it is necessary to carry out a exhaustive differential diagnosis, should consider, among them, the infection for Leishmania, a parasitosis whose incidence is increasing notably in the last years in the immunosuppressed population. We present the clinical case of a 63 year-old patient, immunoincompetent as a consequence of secondary chronic steroid therapy to asthmatic bronchopaty that experiences an uncommon form of visceral leishmaniasis in our area, consistent in multiorganic failure in the context of the development of a macrophage activation syndrome. PMID- 19295976 TI - [Suppurative thrombophlebitis central venous catheterization]. AB - Suppurative thrombophlebitisis a very infrequent complication of the central venous catheterization. The majority of the cases are produced by species of staphylococci. A 22-year-old colombian-woman, student, without toxic habits was admitted because of temporary left astrocytoma (grade II). Nine days after implementing a catheter in the right subclavian vein she presented high fever (39.5 degrees C), shivers, progressive right side neck swelling and odinofagia. She had leukocytosis (26,300 cells/microl) and normal cerebrospinal fluid. After withdrawing the catheter, Staphylococcus aureus was isolated in the tip of the catheter and in four bottles of blood cultures. A neck CT demonstrated expansion and absence of contrast in the right internal jugular vein. The patient evolved satisfactorily with cloxacillin, gentamycin and low molecular weight heparin. PMID- 19295977 TI - [Peritoneal tuberculosis as a complication in a case of unclassifiable myelodysplastic/myeloproliferative disease]. AB - We report the case of a 68-year-old male with a diagnosis of unclassifiable myelodysplatic/myeloproliferative disease (WHO classification), under prolonged steroid treatment and unsuccesful chemotherapy response, who developed progressive asthenia, thoracic pain, minimal efforts dyspnea, and abdominal distension, that initially was suspicious of splenic rupture. Exploratory laparotomy showed multiple peritoneal implants, and a diagnosis of peritoneal tuberculosis was obtained from local biopsy. Definitive diagnosis included a positive result to culture and PCR urine test, together with a possible pleural and splenic tuberculous affectation. Response to tuberculostatic treatment was successful. To the best of our knowledge, this is the first reported case with such characteristics. PMID- 19295978 TI - [Acute renal failure in a young male with cellulitis in the lower leg]. AB - Necrotizing fascitiis due to Streptococcus Pyogenes has a high mortality rate. Detection of the infection before it developes to the streptococcal toxic shock syndrome is quite challenging and its one of the main goals of its management because at this final stage the treatment is in most of the cases ineffective. In a secuence of events of the progression of the infection to shock, renal failure occurs before hipotension very often. We report the case of a 38-year-old patient affected by a fulminant necrotizing fascitiis by Streptococcus Pyogenes which presented at admission with lower leg cellulitis and acute renal failure. PMID- 19295979 TI - [Optic neuromyelitis. Main differences with multiple sclerosis]. AB - The optic neuromyelitis or syndrome of Devic is an inflammatory and autoimmune illness of the central nervous system. It is characterized by attacks of optic neuritis and myelitis, being able to produce blindness, great neurological disability and even the short term death. Until the moment an effective treatment doesn't exist, the therapy is centred in the treatment of the acute attacks, the medical prevention of the complications and the rehabilitation. This article is a revision of this not very common illness, considering that its prevalence in our country has gone in increase. We compare between the optic neuromyelitis and the multiple sclerosis, being based on the main ones characteristic clinical-epidemic that distinguishes these two pathologies, considered by many clinical variants of oneself illness. PMID- 19295980 TI - [Perioperatory artificial nutrition]. AB - Malnutrition increases post surgical morbimortality, hospital stance and economical costs. Possibilities of nutritional intervention in surgical patients are important. Early enteral nutrition is better than total parenteral nutrition in patients under surgery. Periroperaoty nutritional support must be administrated to patients with severe or middle undernutrition and will be under surgery, during 7-14 days before surgical intervention, if this intervention could be delayed. Total parenteral nutrition will be not used regularly in patients under mayor digestive surgical procedures. Inmunonutrition has been demonstrated useful in surgical patients. Evidence demonstrates that inmunotritional formulas decrease incidence of infections, hospital stance and time of ventilation in patients in UCI wards. New research areas have been explored in this topic area, carbohydrate utility in presurgical patients and probiotic in enteral formulas. PMID- 19295981 TI - [Autoimmunity from the medicine based on Virchow's postulates to the medicine based on the postulates of the molecular biology and molecular epidemiology]. PMID- 19295982 TI - [Pneumocephalus after cocaine inhalation]. PMID- 19295983 TI - [Ileal endometriosis as cause of intestinal obstruction. A case presentation]. PMID- 19295984 TI - [Muscle weakness as first manifestation of panhypopituitarism secondary to empty sella syndrome]. PMID- 19295985 TI - [High intestinal haemorrhage in patient with Schoelin-Henoch purpura]. PMID- 19295986 TI - [Slow progression primary pulmonar leiomiosarcoma]. PMID- 19295987 TI - [Association between the use of antipsychotic drugs and the metabolic disorders in a Spanish adult population setting]. PMID- 19295989 TI - [Prognosis factors in colorectal cancer]. PMID- 19295988 TI - [Emerging zoonosis]. PMID- 19295990 TI - [Analysis of possible influence of synchronous neoplastic lesions on prognosis of resected colorectal cancer]. AB - AIM: To analyze the relationship between synchronous lesions in patients with colorectal cancer and their prognostic value. PATIENTS AND METHODS: We have retrospectively reviewed 369 patients with resected colorectal cancer. We compared the rate of apparently curative surgery, progression and tumoral relapse, development of extracolonic cancer and mortality between patients with and without synchronous cancer. Afterwards, we analyzed the same parameters in colorectal cancer with and without synchronous adenomas. Finally, we repeated the analysis after stratification of cancers in 2 groups according to pTNM staging: 0 I-II stage vs III-IV. RESULTS: We found synchronous adenomas in 54.7% of our patients and synchronous cancers in 7.6%. Follow-up period of groups with and without synchronous lesions were: 70.8 +/- 22.9 and 67.2 +/- 24.5 months (p = 0.55) respectively. Synchronous cancers showed higher mortality: 35.7 vs. 14.4%: p = 0.006; OR = 3.31 (1.33-8.13), higher tumoral progression : 39.3 vs. 19.1%: p = 0.011; OR = 2.75 (1.14-6.56) and higher relapse rate: 10.7 vs. 3.5%: p = 0.096. Stratifying according to stage, patients with stage 0-I-II and synchronous cancer showed worse prognosis: mortality = 27.7 vs. 8.1%, p = 0.019; OR = 4.45 (1.2 15.1), tumoral progression = 27.8 vs. 8.5%, p = 0.02; OR = 4.12 (1.14-14.19), and extracolonic cancer = 16.7 vs. 6.4% p = 0.095. There were no statistical differences between cases with and without synchronous adenomas. CONCLUSIONS: Synchronous cancers showed worse prognosis after resection, with higher rate of tumoral progression and mortality. This difference is focused on the cases diagnosed in stage 0-I-II, not being found in III-IV. The presence of synchronous adenomas doesn't influence prognosis. PMID- 19295991 TI - [Prevalence of metabolic syndrome (MS) in laboral population. The heart of Asepeyo]. AB - BACKGROUND: The high increase of the metabolic syndrome (MS) on the occidental World, is increasing the cardiovascular disease. Since, as the presence of metabolic syndrome suggests the application of preventive measure necessary, We studied, the prevalence of metabolic syndrome and the cardiovascular risk factors (RF) in our laboral population. METHODS: The sample included 345 workers, 191 females and 154 males. All the people passed a laboral check-up at the year 2006. RESULTS: Obesity was presented in 12.5% of the sample, 16.9% in males, 8.9% in females. The prevalence of MS was 7.8%, being higher in males than in females. 57.7% of obese males presented MS and 29.4% of obese females presented MS. The older worker presented higher prevalence of MS. The high blood pressure was the factor more prevalent, in worker with MS. All the factors of MS were more prevalent in males, to exception of waist circumference. CONCLUSIONS: 1. Almost 8% of workers presented MS, being higher the prevalence in males. The obesity increased of important manner the MS. The older worker had higher prevalence of MS. 2. The waist circumference associated with all the FR related with MS. PMID- 19295992 TI - [Brain abscess. Evaluation of prognostic factors: does the use of antibiotic prescribing protocols improve outcome?]. AB - BACKGROUND: the aim of this study was to evaluate prognostic factors in brain abscess (AB) and influence of management with antibiotic prescribing protocols (APP). PATIENTS AND METHODS: observational study of a cohort of non-paediatric patients with BA admitted at a 944-bed hospital (1976-2005). Data collection from clinical records has been done according to a standard protocol. We analysed epidemiological, clinical, radiological, microbiological and laboratory data associated with mortality. From 1976 to 1983 (Period I), antibiotic treatment was not done according to any internal APP; from 1983 (Period II), antibiotic management was done according to a APP designed by infectious diseases specialists and neurosurgeons. Predictors of mortality were identified by univariate analysis. The influence of the use of APP in outcome was assessed. RESULTS: 104 patients with BA were included (mean age 45 years; range 12-86); presumed primary pathogenic mechanism of BA was identified in 89%; microbiologic diagnosis was made in 76%. Overall mortality was 16.3%. Factors statistically associated with higher mortality were: age > 40 years, ultimately fatal underlying disease, acute severe clinical condition at the onset of BA, altered mental status and inadequate empirical treatment; 33 patients were treated in Period I and 71 in Period II; no statistically significant differences were found between epidemiological, clinical, radiological or microbiological characteristics of the groups except for mean age (> 40 years in 36% and 62% respectively in Period I and II). Rates of resolution of BA were 60 vs. 77.4% (p < 0.05); relapses 21 vs. 7% (p < 0.05) and mortality 18 vs. 15.4% (p > 0.05), in Period I and II respectively. CONCLUSIONS: main prognostic factors associated with mortality in patients with BA are age, rapidly fatal underlying disease, acute severe clinical condition at the onset of BA, altered mental status and inadequate empirical treatment. Empiric treatment according to APP was associated with greater resolution and lower relapse rates. PMID- 19295993 TI - [Liver abscess: retrospective review of 68 cases]. AB - OBJECTIVE: To assess clinical and laboratory features of patients with liver abscesses (LA), and determine prognostic features. METHODS: We performed a retrospective analysis of medical records of patients receiving a diagnosis of LA in the Hospital Severo Ochoa, between 1989-2005. RESULTS: We were able to find 68 patients: 39 males and 29 females; the incidence amounts 26 cases/100,000 hospital admissions; mean age 63 years. A biliary source accounted for 37%, 16% were of portal origin, 7% were ascribed to hematogenous dissemination, 4% direct inoculation during a procedure and no cause could be found in 35%. Liver ultrasonography allowed diagnosis in 43% of cases, and CT scan un 57%. Sixty-two percent of LA were larger than 3 cm in diameter; 28% of cases had multiple abscesses. Cultures of abscess fluid were positive in 71%, and blood cultures in 52%. Globally, we were able to isolate the causal microorganism in 73.5% of cases. Hundred percent of patients received antimicrobials, 56% had percutaneous drainage performed and 25% were surgically managed. There were complications in 13%, 9% suffered recurrences and we found a 19% mortality rate. CONCLUSIONS: LA has an ill-defined clinical picture. A history of neurological disease or abdominal tumor, and multiple LA are associated with an increased complication rate. Following factors correlated with increased mortality: Age-adjusted Charlson's morbidity index > or =5; Quick index < 60% and development of complications. Drainage indication has to be individualized. PMID- 19295994 TI - [Cost effectiveness analysis of varenicline (Champix) for the treatment of smoking in Spain]. AB - OBJECTIVE: To analyse the efficiency of varenicline compared with bupropion, NRT (nicotine replacement therapy) and no pharmacological treatment in Spain. METHODS: A Markov model was developed to analyse the health and economic consequences of smoking cessation therapies. The transition probabilities were taken from published studies. The model allows cost effectiveness analyses for different time frames (10 years, 20 years and life time). Outcomes are measured in terms of incremental life years gained (LYG) and QALYs. Pharmacological costs and costs of medical visits with varenicline and bupropion were considered. Treatment costs of smoking associated morbidity were taken from Spanish studies. RESULTS: The analyses were done under the perspective of the National Health System, discounting costs and health benefits at 3%. The life time cost effectiveness analysis shows that varenicline dominates all other smoking cessation interventions (more effective at a lower cost). This is due to the higher efficacy of varenicline associated with a reduction in smoking related morbimortality, which, in the long term, accounts for health care cost savings that overcome the extra cost of varenicline. Even when shorter timeframes are considered (20 years), vareniclin is cost-effective in comparison with any other alternative. CONCLUSIONS: Varenicline is a dominant option (more effective at a lower cost) compared with all other smoking cessation treatments when the timeframe is the life span of the patient. Varenicline is cost-effective even when shorter timeframes are considered (20 years or more), with an estimated incremental cost per QALY far bellow any threshold commonly accepted in our environment. PMID- 19295995 TI - [Gastroesophageal reflux as a cause of vocal dysfunction]. AB - Vocal cord dysfunction (VCD), is characterized by a paradoxical adduction of the vocal cords during inspiration, and occurs predominantly in young women. Common symptoms are cough, wheezing, episodic dyspnea, and inspiratory stridor. The true incidence and course of the disease are unknown, and it is usually self limited. It can coexist with, or mimic refractory asthma. Psychological disorders were thought to be the principal cause, subsequently multiple organic diseases have also been reported, like gastroesophageal reflux disease (GERD). Diagnosis is made by clinical suspicion and direct observation. The Gold standard for diagnosis is laryngoscopy with visualization of the paradoxical motion of the vocal cords when the patient is symptomatic. Speech therapy and psychotherapy have been used extensively without any prospective study. We report two cases of VCD associated with GERD, both with excellent respond to treatment. PMID- 19295996 TI - [Diagnosis of the spontaneous subarachnoid hemorrhage with short bleeding or long evolution: report of one case]. AB - Subarachnoid hemorrhage implies the presence of blood within the subarachnoid space from some pathologic process. The initial study of choice is an urgent Cranial Computed Tomography scan, but its sensitivity declines with time. So that it is recommended that patients with severe sudden headache but normal Cranial Computed Tomography scan, should have a lumbar puncture performed, more than 12 hours after the onset of symptoms, to rule out subarachnoid hemorrhage. The methods for distinguishing among traumatic lumbar puncture and true Subarachnoid hemorrhage include the erythrocyte level, the "three tube test", D-dimer assay and ferritin in cerebrospinal fluid. But the best technique is the xanthochromia o yellow-to-orange cerebrospinal fluid supernatant, measured spectrographically. We report a case of a young woman with a subarachnoid hemorrhage diagnosed by xanthochromia after 18 days after the onset of bleeding. PMID- 19295997 TI - [Severe ocular symptoms as an initial manifestation of superior vena cava syndrome]. AB - Superior vena cava syndrome is obstruction of blood flow through the superior vena cava (SVC). It is most commonly caused by neoplasm, especially lung cancer. An obstructed SVC initiates collateral venous return to the heart from the upper half of the body and the classic symptoms and signs become obvious. Superior vena cava sindrome is a rare cause of ocular manifestations. We report a case of periorbital swelling, conjunctival edema an orbital proptosis in a male patient presenting malignant superior vena cava obstruction. PMID- 19295998 TI - [Autoimmune pancreatitis. Related to a case]. AB - Autoimmune pancreatitis is an entity that has recently been included in the differential diagnosis of chronic pancreatitis-pancreatic mass. The presence of diffuse, irregular Wirsung's duct stenosis, together with high levels of IgG4, the existence of certain autoantibodies and lymphoplasmacytic infiltration of the pancreas are key factors in the diagnosis. An early diagnostic suspicion and multidisciplinary collaboration are vital in these cases to avoid unnecessary surgery as the differential diagnosis is established with such entities as pancreas cancer, chronic pancreatitis, primary biliary cirrhosis or primary sclerosing cholangitis. Autoimmune pancreatitis has an excellent response to steroid therapy, with complete resolution of clinical, analytical and radiological parameters, as occurred in our patient. The very few cases reported so far suggest the need for the elaboration of long-term follow-up studies to determine the prognosis and frequency of its association with other diseases. PMID- 19295999 TI - [Immune response in optic neuromyelitis]. AB - The Optic Neuromyelitis is an inflammatory and autoimmune illness of the central nervous system. Presently work is carried out a revision of the different mechanisms involved in the pathogenesis of the Optic Neuromyelitis, the paper of the eosinophils is analyzed, of the antibodies against own antigens and of the regulatory T cells in the illness. In the Optic Neuromyelitis is very important the humoral response, the illness exists it is characterized by the immunocomplex deposit, activation of the complement, production of antibodies against proteins of the myelin and eosinophils recruitment in the lesions. It also exists an increase of the expression of chemokines receptors like the CCR3, specific of TH2 cells; the illness is associate predominantly to a TH2 response. PMID- 19296000 TI - [Neoplastic angiogenesis]. AB - Neoplastic angiogenesis is an essential process in the progressive growth of neoplasms and the production of metastasis. Angiogenesis consists of a series of linked and sequential steps that ultimately leads to the development of a neovascular blood supply to the tumor mass. VEGF has got an essential role in neoplastic angiogenesis, therefore it is an important target in the treatment of neoplasms. Bevacizumab, a humanized monoclonal antibody, inhibits VEGF, and may also improve the delivery of chemotherapy to the tumor mass. Multi-kinase ihibitors (sorafenib and sunitinib) are orally administered small-molecules, that inhibit different receptors (essentials in the neoplastic angiogenesis), such as the VEGFR or PDGFR. These agents are useful in the treatment of advanced renal cell carcinoma, and are under investigation in several tumors. PMID- 19296001 TI - [Rhodococcus equi pulmonary infection in a HIV-infected patient and radiological worsening following treatment: Case report]. PMID- 19296002 TI - [Treatment of tetanus with intrathecal baclofen]. PMID- 19296003 TI - [Drugs and serotonin syndrome]. PMID- 19296004 TI - [Primary pulmonary lymphoma: an infrequent cause of pulmonary mass]. PMID- 19296005 TI - [Primary bacteremias caused by Pasteurella multocida]. PMID- 19296006 TI - [Review of eleven cases of erythema induratum at a second level hospital]. PMID- 19296007 TI - [Pneumonia and lumbar spondylodiscitis as complications of Staphylococcus aureus's bacteriemia]. PMID- 19296008 TI - [Giant cell arteritis: and uncommon cause of chronic cough]. PMID- 19296009 TI - Penile strangulation: report of two unusual cases. AB - We report two cases of penile strangulation that presented to our emergency department. In the first case, a 60-year-old man, the object of strangulation was a metallic ring that was extricated using an orthopaedic cutter in the operating theatre. The patient recovered uneventfully. In the second case, a 77-year-old man, the object of strangulation was a plastic bottle, which was extricated using surgical instruments in the emergency department, but the patient subsequently developed postobstructive diuresis. The first case illustrates the difficulty that may be encountered in this delicate yet urgent situation, while the second case reports a rare complication. PMID- 19296010 TI - Prolonged foreign body impaction in the oesophagus. AB - It is not unusual for a foreign body to be swallowed and become lodged in the oesophagus. It is, however, very unusual for such a foreign body to remain lodged for a period of six months. This particular case, a 37-year-old man, is interesting because of the length of time the foreign body, a denture, remained in the oesophagus without complications, its successful removal and the nature of the foreign body, which is prone to cause complications on prolonged stay or during removal. PMID- 19296011 TI - PET/CT localisation of a scapular haemangiopericytoma with tumour-induced osteomalacia. AB - Oncogenic osteomalacia, or tumour-induced osteomalacia (TIO), is a rare paraneoplastic syndrome characterised by hypophosphataemia, phosphaturia, inappropriately low serum levels of 1,25-dihydroxyvitamin D for hypophosphataemia. TIO is caused by mesenchymal tumours that secrete phosphaturic substances, leading to increased renal wasting of phosphates. These tumours are very small in size and grow slowly. Localisation of these tumours has always been difficult with the available biochemical and imaging techniques. At times, despite all efforts, the tumour could not be localised. We report our experience with a 42-year-old woman with TIO where whole-body magnetic resonance imaging could not localise the tumour, a scapular haemangiopericytoma. PET/CT was helpful in the localisation of the tumour which, when surgically removed, resulted in the normalisation of biochemical parameters with clinical improvement. PMID- 19296012 TI - Carotid body tumours: three case reports. AB - A carotid body tumour is a rare presentation of an extra-adrenal pheochromocytoma. They commonly present with asymptomatic neck masses, and careful preoperative evaluation is required to find out the functional and vascular status of these tumours. We report three cases of carotid body paraganglioma, their evaluation, management and the final outcome. PMID- 19296013 TI - Skin and thigh muscle metastasis from papillary thyroid cancer. AB - A 44-year-old man with total thyroidectomy for papillary thyroid carcinoma (PTC) performed at the age of 38 years was referred for mediastinal metastases demonstrated on chest radiograph and high serum thyroglobulin (Tg 328 ng/ml). Computed tomography revealed mediastinal lymph node enlargement with left bronchial compression and reduction of bronchial diameter, as well as two metastases in the left lung. Bronchoscopic biopsy findings showed a poorly differentiated PTC, while a whole body scan after 131-Iodine therapy demonstrated uptake in the right subclavicular region. External beam radiotherapy and chemotherapy yielded no benefit; the neoplasm was aggressive, diffuse and experienced fast growth, leading to the formation of metastases also at unusual sites, such as the skin and thigh muscle. The patient died from a brain metastasis. We report a rare case of PTC metastasis with a poorly-differentiated component in a young patient. Rapid and diffuse metastases also to unusual sites led to death eight years after the initial diagnosis and treatment. PMID- 19296014 TI - Mania as a presentation of primary hypothyroidism. AB - Hypothyroidism is a common problem in clinical practice, with diverse manifestations. Neuropsychiatric problems include affective disorders, disturbances in cognition and psychosis. Mania is commonly associated with hyperthyroidism. Only a few selected case reports mention mania as a presenting feature of hypothyroidism. We report a case of mania with psychotic symptoms in a 47-year-old woman who had no previous history of psychiatric disorder. She had signs of florid hypothyroidism. She required both antipsychotic drugs and thyroxine replacement for the amelioration of her symptoms. The report is followed by a brief review of the literature on mania as a clinical presentation of hypothyroidism and its probable pathogenesis. One has to have a high index of suspicion of underlying organic causes in patients presenting with depression, psychosis or cognitive disorders. PMID- 19296015 TI - Congenital adrenal hyperplasia 11beta-hydroxylase deficiency: two cases managed with bilateral adrenalectomy. AB - This series describes two patients with congenital adrenal hyperplasia due to 11beta-hydroxylase deficiency. The first patient, a ten-year-old with XX genotype, reared as a male, presented with resistant hypokalaemia and hypertension. The second patient, a 23-year-old with XY genotype, presented with bilateral adrenal masses and resistant hypertension. Both the patients were offered bilateral adrenalectomy. These two patients are described with a discussion on the role of bilateral adrenalectomy in the management of difficult cases of congenital adrenal hyperplasia. The association of myelolipoma and testicular rests with this condition is also discussed. PMID- 19296016 TI - Pheochromocytoma and haemophilia: an unusual combination. AB - We report pheochromocytoma and haemophilia occurring in a 19-year-old South Indian man. To the best of our knowledge, this case is the first of its kind to be reported in the medical literature. The patient had bilateral adrenal pheochromocytomas with an extradrenal pheochromocytoma on the left side, and was successfully operated on after optimal preoperative blood pressure control and factor VIII support. PMID- 19296017 TI - Rheumatic mitral stenosis with epistaxis as an initial symptom. AB - Epistaxis is a frequent presentation in the emergency department and often causes significant anxiety among patients and their relatives. We report a 30-year-old man with mitral stenosis presenting with epistaxis as the initial symptom. He had no other symptoms related to mitral stenosis prior to this episode. Epistaxis is a very unusual initial presentation of rheumatic mitral stenosis. PMID- 19296018 TI - Severe left anterior descending artery stenosis with proximal arteriovenous malformation presenting as acute myocardial infarction. AB - Congenital coronary artery anomalies are rare, with an incidence of about 0.06 1.3 percent of all patients undergoing cardiac catheterisation. They are commonly asymptomatic, but potentially serious lesions may lead to myocardial ischaemia, infarction and/or sudden cardiac death. The occurrence of a concomitant stenotic lesion is exceedingly rare. We report an 80-year-old man who presented with acute anterior myocardial infarction. Coronary angiography revealed severe proximal left anterior descending (LAD) and arteriovenous malformation (AVM) from the first septal branch of the LAD. The LAD stenosis and the AVM were successfully treated with two Jomed covered stents. PMID- 19296019 TI - Schizencephaly associated with bipolar II disorder. AB - A 55-year-old man with congenital hemiparesis of the right side, three episodes of generalised tonic-clonic seizure at 16 years of age, and two episodes of severe depression and two episodes of hypomania in the past, presented with severe depression with psychotic symptoms. Computed tomography of the brain showed a grey matter-lined cerebrospinal fluid-filled cleft in the left cerebral hemisphere, involving the temporoparietal region. He was diagnosed to have bipolar II disorder, and was currently severely depressed with psychotic symptoms and schizencephaly. He improved with sodium valproate 1,000 mg/day, quetiapine 450 mg/day and escitalopram 20 mg/day after three weeks without any emergent side effects, and was maintaining well at three months follow-up. Although uncommon, schizencephaly may be considered as one of the differentials in cases of bipolar disorder along with congenital hemiparesis, mental retardation and/or seizures; and neuroimaging should be done to confirm the diagnosis. PMID- 19296020 TI - Double suicide attempt. AB - A suicide pact is an agreement between two or more people to kill themselves. They represent 0.6-4.0 percent of all suicides, with the vast majority being double suicides. Double suicides are quite rare and are generally seen in old, married couples. We present a double suicide pact involving two young brothers aged 20 and 22 years. Using these two detailed cases and other cases reported in the literature, some general observations on the psychodynamics of suicide pacts are discussed. This is the first study in the literature on a suicide pact made by two brothers. PMID- 19296021 TI - Lumbosacral transitional vertebra: clinical and forensic implications. AB - The identification of skeletal remains is one of the classic problems faced by forensic experts. Congenital and acquired malformations can prove to be an important tool for identification, provided antemortem records are available. A lumbosacral transitional vertebra is one such rare congenital anomaly that has clinical and medicolegal implications. We report a case of unilateral lumbosacral transitional vertebra, detected during medicolegal examination of skeletal remains that were recovered from a forested area in coastal Karnataka, India. The congenital anomaly of the transitional vertebra helped in the forensic identification of the deceased. PMID- 19296022 TI - Variation of the axillary arch muscle with multiple insertions. AB - Axillary arch muscles have been described as having variable and sometimes multiple insertions. We report a 90-year-old female cadaver with an axillary arch muscle that originated from the latissimus dorsi and was inserted into the pectoralis major, pectoralis minor and coracoid process. Recognising that axillary arch muscles can be present in such complex forms is important in clinical practice. PMID- 19296024 TI - Preparing effective tables. AB - Tables are used in scientific papers to present information in a concise and efficient manner. Tables are able to provide information at any desired level of detail and precision. Presenting data in a table rather than as part of the text is also an effective way to reduce the length of the text. This article provides some basic guidelines to assist authors in preparing clear and effective tables for their papers. PMID- 19296025 TI - Factors affecting the predictability of SRK II in patients with normal axial length undergoing phacoemulsification surgery. AB - INTRODUCTION: This study aims to evaluate the factors affecting the accuracy of biometry assessment using the SRK II formula in predicting the refractive outcome after uneventful cataract surgery by phacoemulsification for eyes within the normal range of axial lengths. METHODS: A retrospective review of 100 consecutive cases of uncomplicated phacoemulsification surgery performed by two surgeons from January to September 2005 in a single centre was done. Eyes with axial length greater than or equal to 22.5 mm and less than 24.5 mm were included in the study. The error of prediction was calculated by actual postoperative refractive error minus the refraction target. A comparison was made between the low predictability (prediction error more than 0.50 dioptres [D]) and high predictability (prediction error less than or equal to 0.50 D) groups of eyes. Statistical analysis was performed. RESULTS: A total of 267 case notes were reviewed, of which 100 met the inclusion criteria. The mean age was 68.3 years, with a mean keratometric reading of 43.94 +/- 1.27 D and mean axial length of 23.38 +/- 0.51 mm. The mean IOL power used was 21.77 +/- 1.50 D. The mean error was +0.25 +/- 0.67 (range -1.58 to +1.80) D, with the standard error of mean 0.669. 45 percent of the patients were within 0.5 D of the predicted refractive error and 83 percent were within 1.0 D. There was no difference in preoperative corneal astigmatism, mean keratometry, axial length, age or gender of the patient, laterality of the operated eye and intraocular lens power between the low and high predictability groups. There was a negative correlation between the axial length and prediction error. CONCLUSION: SRK II is reliable in the prediction of the refractive outcome in normal axial length eyes. The findings in our study are comparable to those found in previous studies. We found a negative correlation between axial length and prediction error, even within the range of normal axial length eyes. PMID- 19296026 TI - Age-related macular degeneration in Singapore. AB - INTRODUCTION: This study aimed to describe the morphology of age-related macular degeneration (AMD) as well as to obtain an estimate of the population-based incidence rate in Singapore. METHODS: This is a retrospective hospital-based study of AMD cases seen in 1991 and 1992 at the Singapore National Eye Centre (SNEC), a tertiary eye centre. All case notes recorded with the International Classification of Diseases, ninth revision, clinical modification code '362.5', which is 'degenerations of the macula', were retrieved and analysed. Only case notes of patients who were aged 50 years and above and with documented AMD were included in the study. RESULTS: There were 41 (21.8 percent) patients with drusen alone, 39 (20.7 percent) with dry AMD and 108 (57.5 percent) with exudative AMD. The morphology of the disease was similar among the Chinese and non-Chinese and there were no gender differences. A significant majority of patients with dry and exudative AMD had legal blindness at presentation (p-value is less than 0.0001). Notably, 27 (33.3 percent) patients with exudative AMD had improved vision with time. In comparison, the majority of patients with dry AMD or drusen alone tended to have the same or worsening visual acuity over time. The two-year SNEC hospital incidence rate of AMD in 1991-1992 was 0.38 percent or equivalent to 3.8 per 1,000 new cases seen at SNEC. The estimated population-based incidence rate of exudative AMD was 0.02 percent. CONCLUSION: The population incidence of exudative AMD is lower but comparable to the Western population. Patients with exudative AMD tend to have poorer vision as compared to patients with geographical atrophy or drusen. The proportion of exudative AMD to geographical atrophy appears to be higher than in the West. PMID- 19296027 TI - Breast cancer knowledge and screening practices: a survey of 1,000 Asian women. AB - INTRODUCTION: The incidence of breast cancer in Asia is rapidly rising. Knowledge and perception often influence attendance at screening programmes. However, there has not been any survey to assess the level of knowledge of breast cancer in an Asian population. Singapore has a multiracial population and is the only Asian country with a national screening programme. We conducted a survey on 1,000 women to assess their level of knowledge and screening practices. METHODS: A self administered questionnaire was used, where one point was given for a correct answer and zero for an incorrect / "not sure" response. The maximum knowledge score was 19. The women were also surveyed for their screening practices. Points were not assigned to questions on practice. RESULTS: The response rate was 100 percent. The mean score was 11.4 and the median was 12 (range 0-19). The scores were high for general knowledge and disease progression, but poor for risk factors, screening, symptoms and treatment. Several myths and misconceptions were prevalent. Only 53 percent did regular breast self-examination (BSE), while 57 percent of women aged 40 years and above had gone for a screening mammogram. Increasing age, Malay race, lower educational level, lower socioeconomic class and not knowing anyone with breast cancer were significant independent predictors of poor knowledge. Higher educational level and knowledge scores were significant independent predictors of BSE practice and screening mammogram attendance. CONCLUSION: Knowledge affects practice. Public education is required to correct misconceptions and focus on women with poor knowledge. This would help to improve screening attendance rates. PMID- 19296028 TI - Chemopreventive efficacy of curcumin and piperine during 7,12 dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis. AB - INTRODUCTION: Oral carcinoma accounts for 40-50 percent of all cancers in India. Tobacco chewing, smoking and alcohol consumption are the major risk factors associated with the high incidence of oral cancer in India. Our aim was to investigate the chemopreventive potential of curcumin and piperine during 7,12 dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch carcinogenesis. METHODS: Oral squamous cell carcinoma was developed in the buccal pouch of Syrian golden hamsters, by painting them with 0.5 percent DMBA in liquid paraffin, three times a week for 14 weeks. The tumour incidence, tumour volume and burden were determined in the buccal pouches. The status of phase II detoxification agents, lipid peroxidation and antioxidants were estimated by specific colorimetric methods. RESULTS: We observed 100 percent tumour formation in DMBA-alone painted hamsters. Disturbances in the status of lipid peroxidation, antioxidants and phase II detoxification agents were noticed in DMBA-alone painted hamsters. Oral administration of curcumin (80 mg/kg body weight) and piperine (50 mg/kg body weight) to DMBA-painted hamsters on alternate days to DMBA painting for 14 weeks completely prevented the formation of oral carcinoma. Also, curcumin and piperine restored the status of lipid peroxidation, antioxidants and detoxifying agents in DMBA-painted hamsters. CONCLUSION: The chemopreventive efficacy of curcumin and piperine is probably due to their antilipidperoxidative and antioxidant potential as well as their modulating effect on the carcinogen detoxification process. PMID- 19296029 TI - Risk factors for gastric cancer in South India. AB - INTRODUCTION: Stomach cancer is the third most common cancer in South India. A higher incidence has been reported from certain states in northern India, where potential risk factors have been identified. Similar data is available only to a limited extent from southern India. The aim of this case-control study was to evaluate the effects of lifestyle habits and dietary factors on the risk of gastric cancer in South Indians. METHODS: A hospital-based case-control study of matched pairs was conducted in Chennai, India, from 2002 to 2006 in a large tertiary care referral centre in South India. We studied 89 gastric cancer patients and 89 age- and gender-matched healthy controls of the same socioeconomic status. All subjects were interviewed face-to-face by a trained interviewer using a structured questionnaire to collect data about lifestyle habits, such as cigarette smoking, alcohol consumption, tobacco chewing and dietary factors, with special attention to known factors like salted fish, smoked and pickled foods as well as intake of vegetables and fruits. RESULTS: The response rate was 100 percent. There were 64 male and 25 female patients. The male to female ratio was 2.6:1. The demographic characteristics were similar in the case and control populations. Less than ten percent of patients were below the age of 30 years. Approximately 50 percent were between 30 and 60 years of age, and the rest were over 60 years of age. Multivariate logistic regression models indicated that alcohol consumption (odds ratio [OR] 2.3, 95 percent confidence interval [CI] 1.1-4.9, p-value is 0.04) and consumption of pickled food (OR 1.8, 95 percent CI 1.2-3.9, p-value is 0.05) are independent risk factors for the development of gastric cancer. A protective effect of the consumption of pulses (OR 0.4, 95 percent CI 0.2-0.9, p-value is 0.05), showing a 55 percent reduction in risk, was also identified; this could be of use for possible control and prevention of this cancer. Tobacco chewing and cigarette smoking did not emerge as high risk factors for stomach cancer. CONCLUSION: The study showed alcohol and pickled food consumption as independent risk factors for the development of gastric cancer, while consumption of pulses were protective. Cigarette smoking did not predict an increased risk of contracting the disease. PMID- 19296030 TI - Neonatal resuscitation programme in Malaysia: an eight-year experience. AB - INTRODUCTION: The neonatal resuscitation programme (NRP) published by the American Academy of Paediatrics and American Heart Association was launched in Malaysia in 1996. This study aimed to review the outcome of NRP in Malaysia during the first eight years. METHODS: Information on basic demographical data and training activities of NRP providers were collected prospectively from NRP instructors from all over Malaysia during the eight years following the inception of the NRP. The national perinatal and neonatal mortality data during the five year period before and eight years following implementation of the NRP were compared. RESULTS: During the eight years following the launch, 14,575 personnel were trained. 40 percent of NRP-certified personnel worked in areas where delivery services were provided, viz. labour room, operation theatre, obstetric ward, emergency department and maternal and child health clinic. There were very few NRP-certified providers working in emergency departments and most of them were medical assistants. Most of the providers working in neonatal intensive care units (NICUs) and labour rooms were nurses while those in paediatric wards were doctors. All NRP-certified doctors working in NICUs and labour rooms obtained full certificates. Only 80 percent of NRP-certified nurses in these two areas obtained full certificates. There was further serial decrease in perinatal mortality and neonatal mortality rates in Malaysia during the years following the launch of the NRP programme. CONCLUSION: The launch of the Malaysian NRP was associated with further improvement in perinatal and neonatal mortality rates. PMID- 19296031 TI - A qualitative study of factors influencing family physicians' prescription of the Written Asthma Action Plan in primary care in Singapore. AB - INTRODUCTION: The Written Asthma Action Plan (WAAP) educates patients on the early recognition and treatment of deteriorating asthma. It has been adopted in Singapore polyclinics and restructured hospitals in the past few years as recommended by the Singapore National Asthma Programme. Local asthma patients can choose to be treated by family physicians at public polyclinics or by private general practitioners (GPs). In a previous study, half of the physicians in primary care self-reported that they formulated an asthma action plan for their patients, but little is known about their actual practices. This qualitative study explored family physicians' views of WAAP and its implementation in primary care in Singapore. METHODS: Qualitative data was obtained for 29 participants from polyclinics, GP clinics or locum doctors at five separate focus group discussions. The data was coded using NVivo-7 software, audited and analysed to identify emergent themes. RESULTS: The participants generally perceived the usefulness of WAAP. While WAAP was widely used in polyclinics, GPs rarely used it. Even in polyclinics, usage varied and was associated with participants' training and practice of WAAP and task substitution to nurses. They perceived that WAAP would benefit only selected motivated patients who could understand and use it. They were hampered by language barriers in educating their multiracial patients and the perceived lack of time related to patient loads in polyclinics. The GPs lacked training and practice, role models and relied on verbal instructions as a personalised form of care delivery. CONCLUSION: Family physicians generally perceived WAAP as useful, but implementation was hampered by lack of training and practice, language barriers and perceived benefits for patients. PMID- 19296032 TI - Nutritional supplement use among university athletes in Singapore. AB - INTRODUCTION: Nutritional supplements used by athletes can be classified as sports food, dietary supplements and ergogenic aids. The aim of this study was to examine the use of such supplements among university athletes in Singapore. METHODS: 82 athletes from 16 sport disciplines completed a questionnaire which sought information on demographical parameters, type of supplements, frequency of use, motivations, knowledge, expenditure and side effects. RESULTS: The prevalence of supplement use was 76.8 percent and 20 different products were used. Each individual consumed a mean and standard deviation of 3.18 +/- 1.7 products over a 12-month period. The mean number of products consumed daily was 2.1 +/- 1.2. Popular products included sports drinks, vitamin C, multivitamins and traditional/herbal preparations such as essence of chicken, bird's nest and ginseng. Before using a product, 65.9 percent sought information, usually from the media, the Internet, coaches and fellow athletes. However, many did not know where to obtain reliable information and 86.4 percent were also unaware that supplementation can have adverse effects. CONCLUSION: Although there is a high prevalence of supplement use in our study population, many do not have accurate information about these products. Hence, there is an urgent need to provide athletes with education and access to scientific and unbiased information. PMID- 19296033 TI - A randomised controlled trial of peer-adult-led intervention on improvement of knowledge, attitudes and behaviour of university students regarding HIV/AIDS in Malaysia. AB - INTRODUCTION: The aim of this study was to investigate the knowledge, attitudes and behaviour of university students regarding acquired immunodeficiency syndrome (AIDS) and the human immunodeficiency virus (HIV). METHODS: A randomised controlled trial of 530 university students was done using peer-adult facilitators. Participants completed a questionnaire before and after the intervention, which was a four-hour group session. A prevention programme was developed by local experts, health educators and peer facilitators. The peer adult-led programme was designed to provide a conceptual model of HIV risk reduction through information, motivational and behavioural skills, a harm reduction module and health promotion theme. RESULTS: The main outcome measured was the level of knowledge, attitudes and behaviour scores. The results suggest that relative to the control group, participants in the intervention group had higher levels of knowledge (30.37 vs. 25.40; p-value is 0.001) and a better attitude (12.27 vs. 10.84; p-value is 0.001). However, there was no difference in the behavioural score (9.47 vs. 9.41; p-value is 0.530). The correlation between the level of knowledge and age and the level of education was found in the intervention group, but not in the control group (p-value is 0.01). Attitude and gender were found to be correlated in the intervention group only (p-value is 0.01). CONCLUSION: Our programme was successful in increasing knowledge and improving attitudes towards AIDS and HIV. However, it did not improve risk-taking behaviour. Peer-adult-led educational programmes for youth using various interactional activities, such as small group discussions, poster activity and empathy exercises, can be successful in changing the prevailing youth perceptions of AIDS and HIV. PMID- 19296034 TI - Factors associated with breastfeeding patterns in women who recourse to health centres in Zahedan, Iran. AB - INTRODUCTION: The practice of breastfeeding, an important need for infants, can be affected by several factors that need to be investigated. This study was designed to survey the patterns and period of breastfeeding and its associated factors in women. METHODS: The study was performed in Zahedan, southeast Iran, in 2004-2005, with a sample of 450 mothers via a questionnaire to collect information about the period of breastfeeding and some important factors concerning both mothers and children. RESULTS: The exclusive breastfeeding ratio obtained from this study was 98 percent. The proportion of breastfeeding in months 1, 3, 6, 12 and 24 after birth were 92 percent, 85 percent, 69 percent, 56 percent and 8 percent, respectively. The median breastfeeding period was 15 months, with a standard error of 1.17. The period of breastfeeding showed a significant relationship with the age of the children and mothers, the mothers' education level, night breastfeeding, breastfeeding to an ill child, breastfeeding during a mother's illness and the frequency of breastfeeding per 24 hours. The Cox regression also confirmed a significant relationship between the period of breastfeeding and the above-mentioned variables, except for mother's education level and breastfeeding to an ill child. CONCLUSION: Young pregnant women need to be aware of the importance of breastfeeding and of avoiding the reduction or interruption of the breastfeeding period. It is recommended that the health authorities incorporate training and education programmes as well as healthcare measures to their family health programmes in order to overcome the problems of insufficient breastfeeding. PMID- 19296035 TI - Comparison of outcome and quality of life: haemodialysis versus peritoneal dialysis patients. AB - INTRODUCTION: Ever since peritoneal dialysis (PD) was introduced as a form of renal replacement therapy, its efficacy and complications have been compared with that of haemodialysis (HD). The aim of this study was to determine the efficacy and outcome of PD in comparison to HD in our region. METHODS: We compared 60 patients on PD with 60 matched patients on HD in Tabriz's Sina Hospital during the period 2004-2006. The technique, patients' survival and quality of life were compared by means of a health-related quality-of-life questionnaire (GHQ-28). RESULTS: There was no significant difference in the mean age and duration of dialysis between patients on PD and HD. Survival of diabetic patients was better with HD than PD, but in non-diabetic patients, there was no difference in the survival rates between the two groups. Among patients on PD, diabetics had a 25 percent higher mortality rate and non-diabetic patients had a three percent higher mortality rate than their corresponding counterparts on HD. In all four axes of the questionnaire, i.e. psychophysical dysfunction, stress and sleep disorders, social dysfunction and major depression, PD patients had lower scores than HD patients (p-values are less than 0.001, less than 0.001, equal to 0.002 and less than 0.001, respectively), indicating that patients on PD had a better quality of life compared to those on HD. CONCLUSION: In this study, technique, patients' survival and their quality of life were better on PD than on HD. However, survival and mortality of diabetic patients on HD were better than those on PD. PMID- 19296036 TI - High-resolution computed tomography features in patients with chronic obstructive pulmonary disease. AB - INTRODUCTION: In recent years, there has been increasing interest in diagnosing various components of chronic obstructive pulmonary disease (COPD) using high resolution computed tomography (HRCT). The present study was undertaken to evaluate HRCT features in patients with COPD. METHODS: 40 male patients with COPD (age 40 years or older) and with a significant smoking history (20 pack-years or more) were included in the study. They were evaluated for HRCT features including vascular attenuation and distortion, mosaic attenuation pattern, directly visible small airways, low attenuation areas of emphysema and measures of hyperinflation of the lungs: tracheal index, sterno-aortic distance, thoracic cage ratio and thoracic cross-sectional area. RESULTS: The tracheal index ranged from 0.46 to 0.94; Saber-sheath trachea was found in 14 patients. The mean thoracic cage ratio at two levels, carina and 5 cm below carina, were 0.69 (range 0.61-0.78) and 0.73 (range 0.62-0.83), respectively. Sterno-aortic distance at carinal level ranged from 1.43 to 4.55 cm, with a mean of 3.00 cm. Directly visible small airways was the commonest finding (36 patients), followed by vascular attenuation (25 patients), mosaic attenuation pattern (16 patients) and vascular distortion (8 patients). Among various subtypes of emphysema, centriacinar emphysema was commonest (16 patients), followed by paraseptal (13) and panacinar emphysema (11). CONCLUSION: There are certain specific HRCT features of emphysema and it is possible on HRCT to identify the subtypes of emphysema, such as centriacinar, panacinar and paraseptal emphysema. Various features of hyperinflation can also be well identified and quantified on HRCT. PMID- 19296037 TI - Fibroblasts in the human vocal fold mucosa: an ultrastructural study of different age groups. AB - INTRODUCTION: An investigation was carried out to determine the morphological characteristics of fibroblasts in two portions of the vocal fold (VF) mucosa, the macula flava (MF) and Reinke's space (RS), of three different age groups: newborns, adults and geriatrics. METHODS: Normal human VF obtained from autopsy cases were included in this study: four from mature newborns; four from middle aged adults; and four from geriatric cases. Fibroblasts in RS and MF were investigated by transmission electron microscopy. RESULTS: The fibroblasts of the MF in both adults and newborns tended to be stellate in shape, with a small nucleus/cytoplasm (N/C) ratio and a well-developed rough endoplasmic reticulum (rER) and Golgi apparatus (GA). Most of the fibroblasts present in RS were oval in newborns and spindle-shaped in adults, with a large N/C ratio and less developed rER and GA. The majority of fibroblasts of the geriatric MF were stellate in shape; while in geriatric RS, the majority of fibroblasts were spindle-shaped with an N/C ratio of 0.5 to 2.0 as in the case of adults. However, the development of rER and GA was less marked in geriatrics than in adults. CONCLUSION: Histological changes of fibroblasts in the VF mucosa are one of the important causes of the change in voice quality with ageing. Furthermore, geriatric changes in the vocal ligament can be attributed to the activities and the presence of ageing processes in fibroblasts of geriatric VF mucosa. PMID- 19296038 TI - Antibiotic susceptibility pattern of Staphylococcus species isolated from telephone receivers. AB - INTRODUCTION: Microorganisms are transferred to everyday objects from the environment and infected individuals. Pathogenic microbes are transmissible from the air, skin, hands and other interpersonal contacts and cause diseases most of the time. This study centres on the microbial assessment of telephone receivers, their ability to transfer bacterial infections and the isolation of these microorganisms from receivers, with an emphasis on Staphylococcus aureus and its antibiotic susceptibility pattern. METHODS: 1,591 isolates were obtained from roadside telephone booths in 16 different locations in the Lagos metropolis. They were all characterised using the Cowan and Steel's manual, after which the antimicrobial susceptibility pattern of the most frequently-occurring isolate was determined. RESULTS: Of all the locations, Oshodi was found to have the largest number of microorganisms, with Akoka having the least. The organisms were Providencia, Klebsiella, Citrobacter, Enterobacter, Proteus, Escherichia, Staphylococcus, Bacillus, Streptococcus, Micrococcus and yeast. Staphylococcus spp. was the most frequently occurring, making up 20.2 percent of the total number of isolated organisms, followed by Bacillus (18 percent), while Enterobacter and Citrobacter were the least common. 44 percent of the total Staphylococcus screened for an antimicrobial susceptibility pattern showed high resistance to most of the antibiotics used. CONCLUSION: This result could be related to the emergence of Staphylococcus-resistant strains, especially in a highly-populated area where there are health and hygiene problems and where drugs are abused. Precautions should be taken to prevent the spread of infectious diseases through the use of public telephones. PMID- 19296039 TI - Ultrasonography of the hand and wrist. AB - Musculoskeletal ultrasonography (US) of the hands and wrist has recently been increasing in popularity. Recent rapid technical advances in the US, such as new ultra-high frequency probes and smaller probe sizes, have led to improved image quality. This, in turn, has accelerated the growth of musculoskeletal US. Known advantages of US are its lack of ionising radiation, noninvasiveness, portability and low cost. Dynamic and real-time assessment and Doppler imaging are additional benefits of this modality, especially in the imaging of the hands and wrist. Superficial structures of the hands and wrist, including the tendons, ligaments, nerves and vessels, are amenable to imaging with high frequency US. In this article, we demonstrate a spectrum of hand and wrist pathology using US, including entrapment neuropathy, inflammatory conditions, traumatic injury and masses. Ultrasound-guided procedures applicable to the hand and wrist are also briefly discussed. PMID- 19296040 TI - Charles Robert Darwin: 200 years old. PMID- 19296042 TI - The PII superfamily revised: a novel group and evolutionary insights. AB - The PII proteins compose a superfamily of signal transducers with fundamental roles in the nitrogen metabolism of prokaryotic organisms. They act at different cellular targets, such as ammonia transporters, enzymes, and transcriptional factors. These proteins are small, highly conserved, and well distributed among prokaryotes. The current PII classification is based on sequence similarity and genetic linkage. Our work reviewed this classification through an extensive analysis of PII homologues deposited in GenBank. We also investigated evolutionary aspects of this ancient protein superfamily and revised its PROSITE signatures. A new group of PII proteins is described in this work. These PII homologues have a peculiar genetic context, as they are associated with metal transporters and do not contain the canonical PROSITE signatures of PII. Our analysis reveals that horizontal gene transfer could have played an important role in PII evolution. Thus, new insights into PII evolution, a new PII group, and more comprehensive PROSITE signatures are proposed. PMID- 19296043 TI - Appraisal of peritoneal cavity's capacity in order to assess the pharmacology of liquid chemotherapy solution in hyperthermic intraperitoneal chemotherapy. AB - BACKGROUND: Cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy (HIPEC) is an aggressive strategy to treat patients presenting peritoneal carcinomatosis from various origins. The aim of this study was to evaluate the correlation between the peritoneal cavity's capacity and the weight, the size and the body mass index (BMI) of patients to see if it would be relevant to adapt the pharmacology of HIPEC based on these factors. MATERIALS AND METHODS: This study included 100 patients who had chest-abdominal-pelvic Computerised Tomography (CAP-CT) for various reasons. They were chosen randomly (53 males; 47 females; age range 19-96; mean 58 years). Weight and height of each of them were recorded with their identity on a model sheet given to nurses accustomed to work in clinical trials, before the CAP-CT. The BMI was then calculated from these two values. All the subjects were scanned with CT (Philips Brilliance 40, Cleveland, USA) and the volume of the peritoneal cavity, the liver and the spleen of each was measured with Centricity PACS LS software or Volume Viewer 2 (AW Suite 2.0 6.5.1 u) software. RESULTS: The rates of correlation between the weight, the size, the BMI and the volume of the peritoneal cavity in which the volumes of the liver and the spleen were removed are 0.674, 0.317 and 0.576, respectively; and those of the weight, the size, the BMI and the volume of the peritoneal cavity without taking into account the volume occupied by the liver and spleen are 0.749, 0.348 and 0.644, respectively. CONCLUSION: The peritoneal cavity's capacity is mainly correlated with weight and the interest to assess the volume of liver and spleen remains questionable in terms of results. PMID- 19296044 TI - Evaluation of milk basic protein supplementation on bone density and bone metabolism in Chinese young women. AB - INTRODUCTION: Milk is a good source of bioavailable calcium compared with other foods. Recent in vitro and in vivo studies have shown that milk whey protein, especially its basic protein fraction (milk basic protein, MBP), contains several components capable of promoting bone formation and inhibiting bone resorption. The objective of this study was to examine the effects of MBP on bone mineral density (BMD) and bone metabolism of healthy young women. METHODS: Eighty-four healthy young women were randomly assigned to three groups: control group, whole milk group or MBP group treated with milk containing 40 mg MBP for 8 months. The bone mineral density of total body, the lumbar vertebrae L2-L4 and the left forearm of each subject were measured by dual-energy X-ray absorptiometry (DEXA) at 0 and 8 months of treatment. Serum indexes of bone metabolism were measured at 0, 3, 6 and 8 months. Eighty-one subjects who completed the study in accordance with the protocol were included in the analysis. RESULTS: Total BMD in all groups significantly increased compared with baseline values. However, no significant difference on the mean rate of gain of total BMD was observed among the MBP group (2.19%), the whole milk group (2.63%) and the control group (1.61%). Serum cross linked N-teleopeptides of type-I collagen (NTx) in MBP group at 8 months and in whole milk group at 6 months were significantly decreased from baseline. There were no significant differences between whole milk group and MBP group; however, after combining the milk groups, NTx had significantly decreased from baseline. No significant increase was observed in serum bone-specific alkaline phosphatase (BAP) in both whole milk group and MBP group. CONCLUSION: No significant effect of MBP on bone mineral density and bone metabolism was observed, but milk supplementation was effective in suppressing bone resorption. PMID- 19296045 TI - Impulsivity-hyperactivity and subtypes of aggression in early childhood: an observational and short-term longitudinal study. AB - This short-term longitudinal study (N = 112) was conducted to explore the concurrent and prospective associations between teacher-reported impulsive hyperactive behavior and observed relational and physical aggression during early childhood (M = 45.54 months old, SD = 9.07). Multiple informants and methods including observational methods (i.e., 160 min per child) were used to assess aggression and impulsivity-hyperactivity. All measures were found to be valid and reliable. Prospective hierarchical regression analyses revealed that impulsivity hyperactivity was associated with increases in observed physical aggression across time, controlling for initial relational aggression and gender. These findings add to the growing developmental psychopathology literature that suggests that distinguishing between subtypes of aggression during early childhood may be important for understanding the course of impulsivity hyperactivity in young children. Implications for practice are discussed. PMID- 19296046 TI - CSF neurofilament protein analysis in the differential diagnosis of ALS. AB - BACKGROUND: Cerebrospinal fluid (CSF) biomarkers have been studied to differentiate between patients with ALS and neurological controls, but not in comparison to clinically more relevant disorders mimicking ALS. METHODS: In this retrospective study, CSF concentrations of various brain-specific proteins were analyzed in patients with ALS (n = 32) and ALS-mimic disorders (n = 26). RESULTS: CSF concentrations of neurofilament light (NFL) and heavy chain (NFHp35), but not other brain-specific proteins, were significantly higher in patients with ALS than in patients with an ALS-mimic disorder, however with maximum sensitivity or specificity of 80%. The mean CSF level of NFHp35 was 781 ng/L in the ALS group vs. 338 ng/L in the ALS-mimic disorders group and for NFL the mean CSF levels were 62 ng/L vs. 24 ng/L. CONCLUSION: Although CSF concentrations of NFL and NFHp35 are higher in patients with ALS, the diagnostic accuracy for differentiating ALS from ALS-mimic disorders seems insufficient. Our results suggest that, in the clinical work-up of patients suspected of ALS, application of CSF analysis alone is limited but may have potential in combination with other clinical and electrophysiological markers. PMID- 19296047 TI - Assessing structure and function of the afferent visual pathway in multiple sclerosis and associated optic neuritis. AB - The afferent visual pathway is commonly affected in MS. Assessment of the afferent visual pathway using clinical, imaging and electrophysiological methods not only provides insights into the pathophysiology of MS, but also provides a method of investigating potential therapeutic measures in MS. This review summarises the various assessment methods, in particular imaging techniques of the visual pathway. Retinal nerve fibre layer (RNFL) thickness is usually reduced following an episode of optic neuritis. Techniques such as optical coherence tomography, scanning laser polarimetry, and confocal scanning laser ophthalmoscopy are used to quantify RNFL thickness. MRI of the optic nerve is not routinely used in the diagnosis of MS or optic neuritis, but is valuable in atypical cases and in research. T2- weighted images of the optic nerve usually show the hyperintense lesion in optic neuritis and gadolinium enhancement is seen in the acute attack. Quantifying atrophy of the optic nerve using MRI gives an indication of the degree of axonal loss. Magnetization transfer ratio (MTR) of the optic nerve provides an indication of myelination. Diffusion tensor imaging (DTI) of the optic nerve and optic radiation provide information about the integrity of the visual white matter tracts. Functional MRI following visual stimulation is used to assess the contribution of cortical reorganisation to functional recovery following optic neuritis. Investigations including logMAR visual acuity, Sloan contrast acuity, Farnsworth- Munsell 100-hue colour vision tests and Humphrey perimetry provide detailed quantitative information on different aspects of visual function. Visual evoked potentials identify conduction block or delay reflecting demyelination. These collective investigative methods have advanced knowledge of pathophysiological mechanisms in MS and optic neuritis. Relevant ongoing studies and future directions are discussed. PMID- 19296048 TI - The effect and mechanism of action of capsaicin on gastric acid output. AB - BACKGROUND: Capsaicin has beneficial pharmacological properties, such as the ability to improve appetite and digestion. However, capsaicin has been reported to suppress gastric acid output, but to increase secretion; no consensus as to its effects on gastric acid output has been reached, and the underlying mechanisms remain to be elucidated. METHODS: Rat gastric lumen was perfused with capsaicin. Basal acid output and gastric acid secretion stimulated by vagal nerve activation and bethanecol, a muscarinic receptor agonist, were measured. After intravenous infusion of calcitonin gene-related peptide (CGRP), the measurements were repeated. The secretion of gastrin, somatostatin, and histamine was measured in isolated vascularly perfused rat stomach after vagal nerve and bethanecol stimulation, and under the influence of capsaicin. RESULTS: Capsaicin administration had no effect on basal gastric acid output, but inhibited acid secretion resulting from vagal stimulation. Capsaicin had no effect on acid secretion resulting from stimulation with bethanecol. Administration of high-dose CGRP inhibited basal acid output and gastric acid secretion from both vagal nerve and bethanecol stimulation. Low-dose CGRP inhibited gastric acid secretion because of vagal stimulation, but had no effect on basal secretion or acid secretion following stimulation with bethanecol. Capsaicin administration inhibited the stimulated gastrin and histamine secretion and reversed the suppression of somatostatin secretion mediated by vagal stimulation. However, capsaicin had no effect on stimulated gastrin secretion, suppression of somatostatin secretion, or stimulated histamine secretion because of bethanecol. CONCLUSIONS: Capsaicin inhibited gastric acid output, and the mechanism underlying this effect appears to involve vagal nerve inactivation. PMID- 19296049 TI - Bovine dentine organic matrix down-regulates osteoclast activity. AB - Physiological root resorption is a phenomenon that normally takes place in deciduous teeth; root resorption of permanent teeth occurs only under pathological conditions. The molecular mechanisms underlying these processes are still unclear. Our previous study showed that osteoclasts cultured on deciduous dentine exhibited a higher degree of resorption and higher levels of cathepsin K and MMP-9 mRNA than osteoclasts cultured on permanent dentine. These results could be because of different susceptibilities to acid and the different organic matrices between deciduous and permanent dentine. Thus, the purpose of this study was to investigate the effect of dentine extracts from bovine deciduous and permanent dentine on osteoclast activity. Osteoclasts, obtained from mouse bone marrow cells co-cultured with an osteoblast-rich fraction in the presence of 1,25 (OH)(2)-vitamin D3 and PGE2, were incubated with or without 0.6 M HCl extracts from bovine deciduous or permanent dentine for 48 h. TRAP positive cell number, TRAP activity, the areas of resorption pits, and mRNA levels of TRAP, v-ATPase, calcitonin receptor, cathepsin K, and MMP-9 were examined. The results illustrated that TRAP activity, the resorbed area, and the mRNA levels of osteoclast marker genes seemed to be suppressed by both deciduous and permanent dentine extracts. These findings indicate that some factors that suppress osteoclast activity are contained in both deciduous and permanent dentine extracts. Although there was no significant difference in osteoclast activity between deciduous and permanent dentine extracts, osteoclasts incubated with permanent dentine extracts tend to exhibit less resorption activity than those incubated with deciduous dentine extracts. However, we could not clearly explain the causes of this. PMID- 19296050 TI - Glomus jugulare tumours: certain clinical and radiological aspects observed following Gamma Knife radiosurgery. AB - INTRODUCTION: Glomus jugulare tumours represent a great therapeutic challenge. Previous papers have documented good results from Gamma Knife surgery (GKS) with these tumours. However, the relationship between clinical improvement and tumour shrinkage has never been assessed. MATERIALS AND METHODS: There were 14 patients, 9 women and 5 men. The mean follow-up period was 28 months (range 6 to 60 months). All the tumours except one were Fisch type D and the mean volume was 14.2 cm(3) (range 3.7-28.4 cm(3)). The mean prescription dose was 13.6 Gy (range 12-16 Gy). RESULTS: None of the tumours have continued to grow. Eight are smaller and 6 unchanged in volume. Two patients with bruit have had no improvement in their symptoms. Among the other 12 patients, 5 have had symptomatic improvement of dysphagia, 4 in dysphonia, 3 in facial numbness, 3 in ataxia and 2 in tinnitus. Individual patients have experienced improvement in vomiting, vertigo, tongue fasciculation, hearing, headache, facial palsy and accessory paresis. One patient developed a transient facial palsy. Symptomatic improvement commonly began before any reduction in tumour volume could be detected. The mean time to clinical improvement was 6.5 months whereas the mean time to shrinkage was 13.5 months. CONCLUSIONS: Gamma Knife treatment of glomus jugulare tumours is associated with a high incidence of clinical improvement with few complications, using the dosimetry recorded here. Clinical improvement would seem to be a more sensitive early indicator of therapeutic success than radiological volume reduction. Further follow-up will be needed. PMID- 19296051 TI - Contemporary management of jugular paragangliomas (glomus tumours): microsurgery and radiosurgery. PMID- 19296052 TI - Microdialysis patterns in subarachnoid hemorrhage patients with focus on ischemic events and brain interstitial glutamine levels. AB - BACKGROUND: This observational microdialysis (MD) study of 33 subarachnoid hemorrhage patients explores brain interstitial levels of glutamine, glutamate, lactate and pyruvate, and their relationship to clinical status and clinical course at the neurointensive care unit. METHODS: The focus was on ischemic events, defined by clinical criteria or by radiology, and the significance of brain interstitial glutamine levels and lactate/pyruvate (L/P) ratio. RESULTS: Eleven out of 12 periods with an ischemic MD pattern, defined as lactate/pyruvate (L/P) ratios exceeding 40, were either related to delayed ischemic neurological deficits (DIND) or CT-verified infarcts, confirming that L/P above 40 is a specific ischemic and pathological MD measure. Poor admittance WFNS grade (WFNS 4 5) patients had lower glutamine at the onset of monitoring than what good admittance WFNS grade (WFNS 1-3) patients had (P < 0.05). Interstitial glutamine increased over time in most patients. A "glutamine surge" was defined as a period where the interstitial glutamine concentration increased at least 150 microM over 12 h. Fifteen patients had a DIND and associated MD patterns were glutamine surges (n = 12) and/or L/P>40 (n = 6). Seven patients received vasospasm treatment; in five of these the only DIND-associated MD pattern was a glutamine surge. Seventy percent of the glutamine surges occurred during ongoing propofol sedation, and there was no association between extubations and glutamine surges. There was no difference in mean glutamine levels during the monitoring period between patients with favorable 6-month outcome and patients with poor 6-month outcome. CONCLUSION: We suggest that an increasing interstitial glutamine trend is a dynamic sign of augmented astrocytic metabolism with accelerated glutamate uptake and glutamine synthesis. This pattern is presumably present in metabolically challenged, but yet not overt ischemic tissue. PMID- 19296053 TI - Interleukin-2 in CD8+ T cells correlates with Banff score during organ rejection in liver transplant recipients. AB - The aim of this study is to compare the histological grading of acute organ rejection according to the Banff score with intracellular interleukin-2 (IL-2) concentrations in cytotoxic CD8+ T cells from peripheral blood samples. 66 recipients after liver transplantation and 20 healthy controls were included into this study. Blood samples of liver transplant recipients were collected beside routine visits or, in case of suspected organ rejection, with additional liver biopsy. For cytometry, the blood cells were stained with CD3, CD8 and intracellular-IL-2. The percentage of cells with detectable intracellular IL-2 was significantly increased in patients with acute rejection (n = 7, P < 0.001, t Test) compared to recipients without rejection. The percentage of cells with detectable intracellular IL-2 (mean +/- SEM) was 7.6 +/- 0.9% in rejection patients, 2.3 +/- 0.22% in stable liver transplant recipients, and 14 +/- 2.99% in healthy controls. Intracellular IL-2 correlates to the Banff score in rejection patients (Spearmans-rho = 0.81, P < 0.05). This cytometric method shows a good sensitivity (71%) with a cut-off based on a high specificity of 95% for histological proven organ rejection in our study cohort. Measurement of intracellular IL-2 in cytotoxic CD8+ T-lymphocytes by flow cytometry correlates very well to the histological grading according to the Banff score and shows a good sensitivity and excellent specificity in acute organ rejection. PMID- 19296054 TI - Predictors of multivitamin use during pregnancy in Brazil. AB - OBJECTIVES: The study aimed at identifying predictors of multivitamin use during pregnancy in Brazil. METHODS: Birth registry data of 1,774 infants at maternity hospitals in Brazil were used. The effects of maternal health and fertility risk indicators, enabling factors and other maternal characteristics on multivitamin use were evaluated both pooled and stratified by African ancestry. RESULTS: About 14% of the women used multivitamins during pregnancy. Number of previous live births, maternal age and education, number of ultrasound exams and year of pregnancy had significant effects on multivitamin use in the group reporting African ancestry. Maternal acute illnesses and education had significant effects on use in the group without African ancestry. Significant geographic variation in multivitamin use was observed in both groups. CONCLUSIONS: The study identifies several risk indicators, health care access and enabling factors that are predictive of multivitamin use with differences by African ancestry. The study highlights the importance of increasing the awareness of women of childbearing age of the benefits of multivitamin use and identifies barriers that need to be addressed to promote use. PMID- 19296055 TI - Results from a dietary intervention study in preschools "Beastly Healthy at School". AB - OBJECTIVE: Applying the Intervention Mapping Protocol, an intervention was developed to assist Belgian preschools in the implementation of a healthy school food policy. In the present study the impact of the intervention on children's food consumption is investigated. METHODS: Teachers and parents from 16 schools (8 intervention; 8 control) were asked to participate in the study. Teachers registered the children's available food and beverages during the morning and afternoon breaks (data of baseline and follow-up was matched for 618 intervention and 445 control children). Parents were asked to complete a food frequency questionnaire on their children's general consumption (308 intervention and 168 control matches).Linear mixed model analyses were used to investigate the intervention effect. RESULTS: Both assessment methods indicate an increased fruit consumption for intervention children in comparison with control children, although the effect was only significant for the parental reported fruit consumption. Additionally the results suggest that the change is mainly due to increased availability at school. No significant associations were found for the other food items (snacks, vegetables and different types of beverages). CONCLUSION: The results indicate that a healthy food policy at school can improve young children's diet. PMID- 19296056 TI - Soluble endothelial protein C receptor levels in healthy population. AB - Increased levels of sEPCR lead to dysfunction of EPCR-mediated coagulation. The aim of this present study was to determine plasma sEPCR levels in a group of Turkish healthy population including both adults and children. The study population consisted of 230 healthy individuals (108 children and 122 adults) having no acute or chronic disease. Plasma sEPCR levels were measured with ELISA. Analysis revealed a bimodal distribution in both groups. There was a negative relationship between sEPCR levels and the age of individuals (r = -0.385, P = 0.0001). The sEPCR levels of children were found significantly higher than that of adults (P < 0.001). This study is the first study to determine the relationship of sEPCR levels and terms of age. Higher levels of sEPCR may suggest a regulation mechanism for the protein C anticoagulation system over the first years of life. Further studies should be conducted to evaluate the physiological importance and molecular mechanism of increased sEPCR levels in children. PMID- 19296057 TI - [Noise in intensive care units. Do the alarms for subspecialties differ]. AB - INTRODUCTION: Cardiovascular monitoring alarms are frequent in intensive care units (ICUs) and lead to noise levels often exceeding 80 dB. The aim of this study was to evaluate if there are relevant differences between ICUs with different subspecialties in the frequency and distribution of alarm signals, their occurrence during the day, the types of alarms and the underlying vital parameters. METHODS: All alarm signals of the cardiovascular monitoring systems from randomly chosen patients at five different ICUs of the university hospital of Regensburg were evaluated. RESULTS: No significant differences between the ICUs regarding the frequency of alarm signals and only slight differences in the time distribution could be recognized (p=0.02). The most frequent alarm signals were from threshold alarms (61%) followed by technical alarms. The majority of alarms generated were related to invasive arterial blood pressure measurement. CONCLUSIONS: The frequency and distribution of ICU alarm signals seem to be comparable on different ICUs. Therefore, implementation of universal concepts for alarm reduction seems to be applicable regardless of the subspecialty of the ICU. PMID- 19296058 TI - [Subungual squamous cell carcinoma. An important differential diagnosis of subungual tumors]. AB - A 63-year-old patient presented with a 12-month history of a subungual tumor on the right middle finger. The patient had had a similar lesion involving his left index finger 5 years ago, which was identified as a subungual squamous cell carcinoma. Ablation of the nail plate, the nailbed and the matrix was performed. Histopathology confirmed another subungual squamous cell carcinoma. The patient received a full thickness skin graft. Subungual squamous cell carcinomas are among the most frequently reported types of subungual malignancies. However, subungual squamous cell carcinomas arising in more than one digit have been reported only rarely. The cause of subungual squamous cell carcinoma has not been clearly identified, they has been associated with radiation, chronic infection, arsenic ingestion, HPV infection and trauma. PMID- 19296059 TI - [Merkel cell carcinoma. Viral genesis and new therapeutic options?]. AB - Merkel cell carcinoma (cutaneous neuroendocrine carcinoma) is a rare, highly malignant, neuroendocrine tumor of the skin with predominance in older patients. The tumor is most often located in the sun-exposed skin of the head, the neck and -as in our patient - the extremities. Notably, the tumor bears a high risk of an early regional lymph node as well as distant metastases. Clinically, only a presumptive diagnosis of Merkel cell carcinoma can be established. The definite diagnosis is made by histological and immunohistological methods. Surgical excision with a safety margin should be combined with sentinel lymph node biopsy. In advanced tumor stages (lymph node or visceral metastasis), a remission can be achieved by different chemotherapy schedules in combination with radiation. Recently, a previously unknown polyomavirus, named Merkel cell polyomavirus (MCV or MCPyV), has been identified in 80% of Merkel cell carcinomas. In the near future, these novel findings could be utilized to distinguish Merkel cell carcinoma from small round cell cancers and could lead to the development of new therapeutic options. PMID- 19296060 TI - [Tuberculosis cutis colliquativa. Even today an important differential diagnosis]. AB - An 82-year-old woman presented with perforating abscesses in the cervical and axillary region over 7 months. Histopathological and extensive microbiological examinations lead to the diagnosis of tuberculosis cutis colliquativa, a form of postprimary cutaneous tuberculosis. Tuberculosis cutis colliquativa can be caused by either contiguous spread from an underlying structure, direct inoculation or hematogenous dissemination. Morphologically it is characterized by a subcutaneous node forming an abscess with secondary perforation of overlying skin. The parotid, submandibular, and supraclavicular regions are the sites most likely to be affected. Today it still remains an important differential diagnosis for an abscess with fistulas. Correlation with histopathologic findings and microbiological examinations including polymerase chain reaction and mycobacterial culture being the most reliable method is required. Standard therapy regimens consist of 2 months of quadruple therapy followed by another 4 months of a continuation phase using a two-drug regimen. PMID- 19296061 TI - Calciphylaxis associated with rheumatoid arthritis: communication of the second case. AB - Calciphylaxis is the tissue sensitivity to calcification, described mainly in patients with chronic renal insufficiency, renal transplant of parathyroid dysfunction. There is only one case described in a patient with rheumatoid arthritis, without renal dysfunction, nor hyperparathyroidism. We present the second case in a woman with inactive rheumatoid arthritis. PMID- 19296063 TI - A case of acute cardiomyopathy and pericarditis associated with methylphenidate. AB - Methylphenidate is a potent central nervous system stimulant that exerts its effects by increasing synaptic levels of dopamine and norepinephrine. It has become key to treating attention deficit-hyperactivity disorder (ADHD) in children and adolescents. As the use of stimulant medications has ballooned in the past decade, so too has awareness of the cardiovascular complications of these drugs. Effects on heart rate and blood pressure as well as tachyarrhythmias have been well described. However, acute cardiomyopathy and pericarditis secondary to methylphenidate use has been rarely reported. We report the case of a 17-year-old male who developed chest pain, elevated cardiac biomarkers, and acute left ventricular dysfunction following a single dose of methylphenidate. The risk of cardiomyopathy in the setting of methylphenidate treatment should prompt further study on the safety of this drug, and lead to ways of identifying those at risk of developing these complications. PMID- 19296062 TI - Bone mineral density at the hip in Norwegian women and men--prevalence of osteoporosis depends on chosen references: the Tromso Study. AB - This study describes bone mineral density (BMD) and the prevalence of osteoporosis in women and men between 30-89 years in an unselected population. BMD was measured in g/cm(2) at total hip and femoral neck by dual-energy-X-ray absorptiometry in 3,094 women and 2,132 men in the 2001 Tromso Study. BMD levels were significantly explained by age and declined progressively in both sexes from middle into old age, with highest decline in women. With osteoporosis defined as a T-score of two and a half standard deviation below the young adult mean BMD, the prevalence at the total hip in subjects above 70 years was 6.9% in men and 15.3% in women, respectively, using the Lunar reference material for T-score calculations. The prevalence increased significantly to 7.3% in men and 19.5% in women, when T-scores were calculated on basis of the young adult mean BMD (age group 30-39 years) in the study population. At the femoral neck, prevalence of osteoporosis increased from 13.5 to 18.5% in men, and from 20.4 to 35.2% in women above 70 years, respectively, depending on how T-scores were calculated. The study highlights the challenges with fixed diagnostic levels when measuring normally distributed physiologic parameters. Although BMD only partly explains fracture risk, future studies should evaluate which calculations give optimal fracture prediction. PMID- 19296064 TI - Direct and indirect fossil records of megachilid bees from the Paleogene of Central Europe (Hymenoptera: Megachilidae). AB - Aside from pollen and nectar, bees of the subfamily Megachilinae are closely associated with plants as a source of materials for nest construction. Megachilines use resins, masticated leaves, trichomes and other plant materials sometimes along with mud to construct nests in cavities or in soil. Among these, the leafcutter bees (Megachile s.l.) are the most famous for their behaviour to line their brood cells with discs cut from various plants. We report on fossil records of one body fossil of a new non-leafcutting megachiline and of 12 leafcuttings from three European sites-Eckfeld and Messel, both in Germany (Eocene), and Menat, France (Paleocene). The excisions include the currently earliest record of probable Megachile activity and suggest the presence of such bees in the Paleocene European fauna. Comparison with extant leafcuttings permits the interpretation of a minimal number of species that produced these excisions. The wide range of size for the leafcuttings indirectly might suggest at least two species of Megachile for the fauna of Messel in addition to the other megachiline bee described here. The presence of several cuttings on most leaves from Eckfeld implies that the preferential foraging behaviour of extant Megachile arose early in megachiline evolution. These results demonstrate that combined investigation of body and trace fossils complement each other in understanding past biodiversity, the latter permitting the detection of taxa not otherwise directly sampled and inferences on behavioural evolution. PMID- 19296065 TI - [Possibilities and risks of the monoclonal antibody alemtuzumab as a new treatment option for multiple sclerosis]. AB - Monoclonal antibodies are of growing interest as treatment options for immune mediated diseases in neurology. As our knowledge of immunological principals increases, we learn to modulate specifically mechanisms of pathogenesis by the use of monoclonal antibodies. It is clearly desirable to improve efficacy in disease treatment without increasing toxicity by using drugs with more specific modes of action. Natalizumab was the first monoclonal antibody approved in the field of neurology for treatment of relapsing remitting multiple sclerosis (MS). Several other monoclonal antibodies are currently under investigation. Alemtuzumab, a monoclonal antibody targeting CD52, is a highly promising agent currently being studied in two phase III clinical trials. In this review, data from the recently published phase II clinical trial in the treatment of early relapsing remitting MS is summarized and analyzed in light of the development of alemtuzumab for MS and its potential role in treating this disease is discussed. PMID- 19296067 TI - [Magnetic resonance imaging and computed tomography findings in lacunar brain infarctions]. AB - Lacunar brain infarctions belong to the most common causes of stroke and are typically located in basal ganglia with embolic, atheromatous or thrombotic etiology. Lipohyalinosis, often as a result of a long history of hypertension, can also be a cause of lacunar lesions. At present more than 20 different lacunar syndromes have been described. However clinical findings may be variable ranging from clinically"silent" to focal neurologic deficits to cognitive impairment. Magnetic resonance imaging (using diffusion-weighted sequences) generally allows diagnosis in most cases of acute lacunar infarcts and is superior to computed tomography especially in cases of additional leucoaraiosis. PMID- 19296066 TI - [Femoral head necrosis]. AB - The epidemiology and pathohistogenesis of avascular femoral head necrosis has still not been clarified in detail. Because the course of the disease runs in stages and over a long time period nearly always culminates in the necessity for a total hip prosthesis, an exact radiological evaluation is of paramount importance for the treatment. There is a need for a common staging system to enable comparison of different therapy concepts and especially their long-term results. In this article the ARCO staging system is described in full detail, which includes all radiological modalities as well as histopathological alterations. PMID- 19296068 TI - [PET-CT for neuroendocrine tumors and nuclear medicine therapy options]. AB - Neuroendocrine tumors (NET) are defined by biochemical characteristics and structures which can be specifically addressed by radioligands for diagnostic imaging as well as radionuclide therapy in nuclear medicine. Somatostatin receptor imaging has been shown to be an important part of the diagnostic process in the management of NET for a long time. In recent years a number of tracers enabling PET-based imaging of somatostatin receptors and amine precursor uptake have been developed. By combining the specific functional information of the PET signal with anatomical information by CT imaging using PET-CT hybrid scanners, primary tumors and metastases can be detected with high resolution and high sensitivity. Compared with conventional indium-111 octreotide scintigraphy PET-CT has a higher resolution and also a lower radiation exposure. In addition, quantification of the tracer uptake allows therapy monitoring. By labelling with therapeutic beta-emitters, such as lutetium-177 or yttrium-90, a systemic internal radiotherapy with somotostatin analogues (peptide radionuclide radiation therapy, PRRT) can be provided as a therapeutic option for patients with unresectable and metastasized neuroendocrine tumors. PMID- 19296069 TI - [Testosterone replacement therapy and prostate cancer. The current position 67 years after the Huggins myth]. AB - Hypogonadism is highly prevalent in the elderly and in men with prostate cancer. Symptoms of hypogonadism, such as depression, lack of libido, and decreased bone mineral density, can significantly impair quality of life. In addition, testosterone plays an important role in erectile preservation and in growth and function of the cavernosal and penile nerves. There are compelling data showing that testosterone replacement therapy (TRT) does not increase the risk of prostate cancer. The literature (four published studies) concerning men treated with TRT after definitive therapy for prostate cancer reports only one biochemical recurrence. Based on these data, physicians cannot really justify withholding TRT from symptomatic patients after they have been successful treated for prostate cancer. This review gives the practising urologist an overview of the latest literature and useful advice on this controversial topic. PMID- 19296070 TI - [Pharmaceutical therapy for bladder dysfunction disorders]. PMID- 19296071 TI - [Treatment of LUTS in BPS. When and when not to administer pills?]. AB - Benign prostatic syndrome (BPS) is considered a diagnosis of exclusion and needs a thorough work-up. One of the pitfalls for a tailored medical treatment scheme is the objective evaluation of benign prostatic obstruction. Characteristics of the various medical therapy options and the multifactorial origin of LUTS in BPH patients imply an individualized approach. LUTS involving mostly urine storage disorders and a small prostate are suitably managed with alpha(1)-receptor antagonists, which may be combined with antimuscarinics if OAB symptoms predominate. Long-term treatment addressing clinical progression may favor combination therapy of alpha(1)-receptor antagonists with 5alpha-reductase inhibitors if prostate size is sufficient. Age, symptom severity at baseline, a large prostate volume, or a high PSA value are indicative of progression. However, combination therapy aggravates side effects, and thus a risk-benefit analysis is essential. The potential of any medication for BPS to treat obstruction is rather low. If deobstruction is the main aim of therapy, medical treatment is not suitable. PMID- 19296072 TI - [Trends in prostate biopsy interpretation]. AB - Driven by the pressure of increasing use of PSA screening, cancer diagnosis based on histopathology has been constantly improved. When appropriate quality standards for obtaining a biopsy specimen and for tissue preparation are upheld, even the smallest tumour foci can be identified with certainty. Efforts to standardize the Gleason grading should also help to render assessments of tumour biology more precise. In addition, subtle diagnosis of precancerous or cancer associated lesions will permit more exact prognoses of existing cancer risk to be made. There are still weak points: reliable recognition of clinically insignificant carcinomas and the danger of overtreatment. Further research activity on this topic is needed. PMID- 19296076 TI - Thiamine in diabetic nephropathy: a novel treatment modality? PMID- 19296078 TI - Decreased levels of metabolic enzymes in pancreatic islets of patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: Glucose-stimulated insulin secretion is defective in patients with type 2 diabetes. We sought to acquire new information about enzymes of glucose metabolism, with an emphasis on mitochondrial enzymes, by comparing pancreatic islets of type 2 diabetes patients with those of non-diabetic controls. METHODS: Expression of genes encoding 13 metabolic enzymes was estimated with microarrays and activities of up to nine metabolic enzymes were measured. RESULTS: The activities of the mitochondrial enzymes, glycerol phosphate dehydrogenase, pyruvate carboxylase (PC) and succinyl-CoA:3-ketoacid CoA transferase (SCOT) were decreased by 73%, 65% and 92%, respectively, in the diabetic compared with the non-diabetic islets. ATP citrate lyase, a cytosolic enzyme of the mitochondrial citrate pyruvate shuttle, was decreased 57%. Activities of propionyl-CoA carboxylase, NADP-isocitrate dehydrogenase, cytosolic malic enzyme, aspartate aminotransferase and malate dehydrogenase were not significantly different from those of the control. The low activities of PC and SCOT were confirmed with western blots, which showed that their protein levels were low. The correlation of relative mRNA signals with enzyme activities was good in four instances, moderate in four instances and poor in one instance. In diabetic islets, the mRNA signal of the islet cell-enriched transcription factor musculoaponeurotic fibrosarcoma oncogene homologue A, which regulates expression of islet genes, including the PC gene, was decreased to 54% of the control level. PC activity and protein levels in the non-diabetic islets were significantly lower than in islets from non-diabetic rodents. CONCLUSIONS/INTERPRETATION: Low levels of certain islet metabolic enzymes, especially mitochondrial enzymes, are associated with human type 2 diabetes. PMID- 19296077 TI - Agreement among type 2 diabetes linkage studies but a poor correlation with results from genome-wide association studies. AB - AIMS/HYPOTHESIS: Little of the genetic basis for type 2 diabetes has been explained, despite numerous genetic linkage studies and the discovery of multiple genes in genome-wide association (GWA) studies. To begin to resolve the genetic component of this disease, we searched for sites at which genetic results had been corroborated in different studies, in the expectation that replication among studies should direct us to the genomic locations of causative genes with more confidence than the results of individual studies. METHODS: We have mapped the physical location of results from 83 linkage reports (for type 2 diabetes and diabetes precursor quantitative traits [QTs, e.g. plasma insulin levels]) and recent large GWA reports (for type 2 diabetes) onto the same human genome sequence to identify replicated results in diabetes genetic 'hot spots'. RESULTS: Genetic linkage has been found at least ten times at 18 different locations, and at least five times in 56 locations. All replication clusters contained study populations from more than one ethnic background and most contained results for both diabetes and QTs. There is no close relationship between the GWA results and linkage clusters, and the nine best replication clusters have no nearby GWA result. CONCLUSIONS/INTERPRETATION: Many of the genes for type 2 diabetes remain unidentified. This analysis identifies the broad location of yet to be identified genes on 6q, 1q, 18p, 2q, 20q, 17pq, 8p, 19q and 9q. The discrepancy between the linkage and GWA studies may be explained by the presence of multiple, uncommon, mildly deleterious polymorphisms scattered throughout the regulatory and coding regions of genes for type 2 diabetes. PMID- 19296079 TI - [Surgical treatment of pathologic fractures of the humerus and femur]. AB - The life expectancy of patients with malignant tumours and the incidence of osseous metastases have increased over the last decades. Operations for skeletal metastases of the extremities represent the most frequent surgery in orthopaedic oncology. The purpose of this study was to evaluate and compare the different operative treatment options for patients with pathologic fractures of the humerus and femur in terms of complications, postoperative recovery, and survival.From 2000 to 2005, 109 patients were surgically treated for pathologic fractures of the humerus (n=19) or femur (n=90). The study group consisted of 60 women and 43 men, with a mean age of 67 years (13-88). Breast carcinoma (36%) was the most common primary tumour, followed by kidney (17%) and bronchial (16%) carcinoma. Of all patients, 75 (73%) had numerous skeletal metastases, and 38 (37%) had visceral metastases.Wide or marginal resection was performed in seven fractures of the humerus and 14 fractures of the femur; intralesional resection was done in seven humeral and 73 femoral fractures; and stabilisation alone was done in five fractures of the humerus and three fractures of the femur. The median survival time for all patients was 6 months (0-102). The survival rate at 1 year was 25% (25% for both humeral and femoral fractures), 15% at 2 years (17% for humeral and 15% for femoral fractures), and 8% at 3 years (16% for humeral and 7% for femoral fractures). The overall complication rate was 11%, and revision surgeries were performed in seven patients (6.4%). The majority of patients (n=65; 60%), especially those with fractures close to the articular joint, were successfully treated with endoprosthetic replacement. Patients with fractures stabilised by intramedullary nails had shorter operating times, a shorter hospital stay, and fewer complications than patients treated with plating systems. Therefore, we recommend intralesional resection of the metastasis and stabilisation with intramedullary devices, supported by bone cement, as the treatment of choice for pathologic fractures of the diaphysis and metaphysis of the humerus and femur. Wide resection should be reserved for selected cases, such as solitary bone metastasis of kidney carcinoma. PMID- 19296080 TI - [Bisphosphonates for malignant bone tumors]. AB - Bisphosphonates (BPs) are able to prevent, reduce, or delay skeletal complications caused by tumors. Since the introduction of BPs, there has been a marked reduction of skeletal events. Today we can choose from a variety of BPs with different potency, efficacy, dosing, and administration regimens, as well as BPs for different indications. For patients with tumors who have a risk of developing osteoporosis induced by the tumor or antitumor treatment, we should think of prevention. In general, BPs are well tolerated. But there are also side effects: flulike syndrome, arthralgias, or, when administered orally, gastrointestinal symptoms. In a number of cases, the dosing regime must be adjusted according to renal function and baseline creatinine clearance. This paper reviews the current evidence-based use of BPs in solid tumors and the recommendations for treatment. PMID- 19296081 TI - [Prognosis-adapted surgical management of bone metastases]. AB - Surgery in metastatic disease to the bone is the most common procedure in orthopaedic oncology. To assess an adequate therapeutic approach we analysed 513 surgeries performed on 453 patients between 1980 and 2005 and reviewed the literature.The most significant factor is the histology of the primary tumour. A biopsy is mandatory to confirm the diagnosis in an unknown primary tumour, especially in cases of solitary lesions. Pulmonary carcinoma has an unfavourable prognosis compared to breast and renal cell carcinoma patients. Radical resection in isolated metastatic disease in renal cell carcinoma reduces the risk of local recurrence and even may result in a long progression-free survival. In breast cancer osseous and visceral dissemination is the most decisive factor for prognosis. An interdisciplinary approach is mandatory in every patient. PMID- 19296082 TI - [New developments in orthopedic oncology]. PMID- 19296083 TI - Outcomes of microfracture in professional basketball players. AB - Surgical treatment for chondral defects of the knee in competitive running and jumping athletes remains controversial. This study evaluated the performance outcomes of professional basketball players in the National Basketball Association (NBA) who underwent microfracture. Data from 24 professional basketball players from 1997 to 2006 was obtained and analyzed. NBA player efficiency ratings (PER) were calculated for two seasons before and after injury. A control group of 24 players was used for comparison. Study group and control group demographics including age, NBA experience, and minutes per game demonstrated no statistical difference. Mean time to return to an NBA game was 30.0 weeks from the time of surgery. The first season after returning to competition PER and minutes per game decreased by 3.5 (P < 0.01) and 4.9 min (P < 0.05), respectively. The 17 players who continued to play two or more seasons after surgery, the average reduction in their PER and minutes per game was 2.7 (P > 0.05) and 3.0 min (P < 0.26), respectively. A multivariant comparison versus controls demonstrated that power rating during the 2 years after surgery decreased by 3.1 (P < 0.01); while minutes per game decreased by 5.2 (P < 0.001). Twenty-one percent (n = 5 of 24) of the players treated with microfracture did not return to competition in an NBA game. On return to competition player performance and minutes per game are diminished. PMID- 19296084 TI - Negative effects of parecoxib and indomethacin on tendon healing: an experimental study in rats. AB - Conventional non-steroidal anti-inflammatory drugs (NSAIDs) and newer specific cyclooxygenase-2 (cox-2) inhibitors are commonly used in muscular skeletal trauma and in relation to orthopedic surgery to reduce the inflammatory response and pain. Studies have indicated that these drugs can have a negative effect on tendon healing in the early proliferative phase, but might be beneficial in the remodeling phase when inflammation might impede healing. Our study was designed to investigate if short-term administration of cox inhibitors after injury or postoperatively might have negative effects on the tendon healing. The right Achilles tendon of 60 rats was cut transversely, a 3 mm long segment of the tendon was removed and left unrepaired. The animals were then given parecoxib, indomethacin or saline intraperitoneally twice daily for 7 days. After 14 days, the animals were euthanized. The transverse and sagittal diameters in the healing area were measured and mechanical testing of the tensile strength of the tendons was performed. We found a significantly lower tensile strength in rats given both parecoxib and indomethacin compared to the control group. Stiffness in the healing tendons was significantly lower in the parecoxib group compared to both the placebo and the indomethacin groups. The transverse and sagittal diameters of the tendons were reduced in both the parecoxib and indomethacin groups. Both parecoxib and indomethacin impaired tendon healing; the negative effect was most pronounced with parecoxib. PMID- 19296085 TI - Elongation of simulated whipstitch post anterior cruciate ligament reconstruction tibial fixation after cyclic loading. AB - Whipstitch-post (WSP) tibial fixation is one of the most widely used and clinically successful methods of soft tissue graft fixation for anterior cruciate ligament reconstruction (ACLR). However, some consider the method prone to laxity. We hypothesized that WSP would have low elongation rates after experimental cyclic loading. Eight cadaveric human semitendinosus and gracilis (ST/Gr) tendons had whipstitches woven into their overlapped ends. The grafts were looped around a metal bar, pneumatically clamped, and cyclically loaded. The adjusted mean experimental graft elongation for the WSP was 1.13 mm with a maximum elongation of 1.64 mm and a standard deviation of 0.32. These values are equivalent to the lowest published cyclic loading tibial fixation elongation data. Whipstitch-post tibial ACLR fixation is biomechanically sound with among the lowest rates of elongation after laboratory cyclic loading. PMID- 19296086 TI - Functional and perceived response to intra-articular hyaluronan injection in patients with knee osteoarthritis: persistence of treatment effects over 5 months. AB - Intra-articular injections of hyaluronic acid are currently indicated as a symptom-modifying treatment for knee osteoarthritis to palliate symptoms and improve function. A series of three to five weekly injections are typically delivered once per week and a period of at least 6 months must pass between consecutive injection series. Little is known about why some patients do not respond to this treatment or about its effects on outcome measures recorded during functional activities. In addition to investigating short-term self-report and measured functional changes, the purpose of this study was to evaluate the persistence of treatment effects over the typical time interval between consecutive treatment series. According to the a priori definition for response to treatment, over two-thirds of patients experienced improved knee function during the typical period between series of intra-articular injection of hyaluronic acid while a one-third did not demonstrate a change in self-report scores. Differences in response were similarly seen between groups in functional tests. Many had immediate benefits and although some did not retain their gains, others experienced a delayed response in the following months. Knee flexion range of motion was related to self-report and tested function, and improvements in range of motion were associated with concurrent improvements in other functional measures across the testing period. A short functional test and knee-specific questionnaire may provide an index that provides an indication as to whether or not a particular patient is likely to respond to hyaluronan injections. Such information may provide valuable input when decisions are made regarding which treatment option is recommended to individuals with knee osteoarthritis. PMID- 19296087 TI - Healing of meniscal tissue by cellular fibrin glue: an in vivo study. AB - Menisci represent fundamental structures for the maintenance of knee homeostasis, playing a key role in knee biomechanics. However, their intrinsic regenerative potential is poor. As a consequence, when a lesion occurs and the meniscus is partially removed by surgery, knee mechanics is subject to dramatic changes. These have been demonstrated to lead often to the development of early osteoarthritis. Therefore, menisci should be repaired whenever possible. In the last decades, tissue engineering approaches have been advocated to improve the reparative processes of joint tissues. In this study, the bonding capacity of an articular chondrocytes-fibrin glue hydrogel was tested as a biologic glue to improve the bonding between two swine meniscal slices in a nude mouse model. The composites were wrapped with acellular fibrin glue and implanted in subcutaneous pouches of nude mice for 4 weeks. Upon retrieval, a firm gross bonding was observed in the experimental samples while none of the control samples, prepared with acellular fibrin glue at the interface, presented any sign of bonding. This was consistent with the histological and scanning electron microscope findings. In particular, a fibrocartilaginous tissue was found at the interface between the meniscal slices, partially penetrating the native meniscus tissue. In order to overcome the lack of regenerative properties of the meniscus, the rationale of using cellular fibrin glue is that fibrin provides immediate stability while carrying cells in the site of lesion. Moreover, fibrin gel is recognized as an optimal scaffold for cell embedding and for promoting fibrocartilaginous differentiation of the cells which synthesize matrix having healing property. These results demonstrated the potential of this model for improving the meniscal bonding. However, further orthotopic studies in a large animal model are needed to evaluate its potential for clinical application. PMID- 19296088 TI - Hypothermic storage of isolated human hepatocytes: a comparison between University of Wisconsin solution and a hypothermosol platform. AB - Until now little is known about the functional integrity of human hepatocytes after hypothermic storage. In order to address this limitation, we evaluated several commercially available hypothermic preservation media for their abilities to protect freshly isolated hepatocytes during prolonged cold storage. Human hepatocytes were isolated from non-transplantable/rejected donor livers and resuspended in ice-cold University of Wisconsin solution (UW), HypoThermosol-Base (HTS-Base), or HypoThermosol-FRS (HTS-FRS) with or without the addition of fetal bovine serum. Cells were stored at 4 degrees C for 24-72 h, and evaluated for hepatocyte viability (trypan blue exclusion, or labeling with fluorochromes), cell attachment, and function. The energy status of hepatocytes was evaluated by measurement of intracellular adenosine 5'-triphosphate. To determine whether the test cells expressed metabolic functions of freshly isolated cells, the activities of major phase I (cytochromes P450, FMO) and phase II (UGT, ST) drug metabolizing enzymes were examined. Although hepatocytes are shown to be satisfactory after 24 h storage in all of the tested solutions, the cell viability, energy status, and xenobiotic metabolism following cold preservation in HTS-FRS was consistently and, in some cases, markedly higher when compared with other systems. The same metabolites for each of the tested substrates were detected in all groups of cells. Moreover, the use of HTS-FRS eliminates the need for serum in preservation solutions. HTS-FRS represents an improved solution compared to HTS-Base and UW for extending the shipping/storage time of human hepatocytes. PMID- 19296091 TI - 17beta-estradiol attenuates vascular contraction through inhibition of RhoA/Rho kinase pathway. AB - We hypothesized that 17beta-estradiol attenuates vascular contraction through inhibition of RhoA/Rho kinase pathway. Rat aortic rings were contracted with cumulative addition of U46619, NaF, KCl or PDBu 30 min after pretreatment with 17beta-estradiol (10, 30, and 100 microM) or vehicle. We measured the amount of GTP RhoA and the level of phosphorylation of the myosin light chain (MLC(20)), myosin phosphatase targeting subunit 1 (MYPT1) and PKC-potentiated inhibitory protein for heterotrimeric MLCP of 17 kDa (CPI17). Pretreatment with 17beta estradiol dose-dependently inhibited the concentration-response curves in response to U46619, NaF or KCl, but not to PDBu. 17beta-Estradiol decreased not only the level of phosphorylation of MYPT1(Thr855) and CPI17(Thr38) as well as MLC(20), but also the activity of RhoA induced by U46619 or NaF. However, 17beta estradiol did not affect the level of phosphorylation of CPI17 induced by PDBu. 17beta-Estradiol attenuates vascular contraction through inhibition of RhoA/Rho kinase pathway. PMID- 19296090 TI - GABA(A) autoreceptors enhance GABA release from human neocortex: towards a mechanism for high-frequency stimulation (HFS) in brain? AB - High-frequency stimulation (HFS) in human neocortical slices induces gamma aminobutyric acid (GABA) release via GABA(A) receptor (GABA(A)R) activation. The mechanism of this effect and the localization of these GABA(A)Rs were now studied. Fresh human neocortical slices were subjected to HFS (130 Hz) in the presence of veratridine (3 microM). As measured by high-performance liquid chromatography, only GABA but not glutamate outflow was affected by HFS/veratridine stimulation. The evoked GABA overflow was abolished by tetrodotoxin and furosemide, suggesting an involvement of action potentials and plasmalemmal chloride gradients. Double immunolabeling showed that GABA(A)Rs are localized on soma and dendrites of GABAergic neurons in the human neocortex. Moreover, in support of a terminal localization of GABA(A)Rs, the K+-evoked [3H] GABA release from synaptosomes was enhanced by the GABA(A)R agonist muscimol (antagonized by GABA(A)R blockers). We conclude that HFS in human brain neocortex leads to a specific increase of GABA release, which is mediated by facilitatory GABA(A) autoreceptors located on soma, dendrites, and axon terminals of GABAergic neurons. PMID- 19296092 TI - Selection of adequate optimization criteria in chromatographic separations. AB - Computer-assisted optimization of chromatographic separations is still a fruitful activity. In fact, advances in computerized data handling should make the application of systematic optimization strategies much easier. However, in most contemporary applications, the optimization criterion is not considered to be a key issue (Vanbel, J Pharm Biomed, 21:603-610, 1999). In this paper, an update of the importance of selecting adequate criteria in chromatographic separation is presented. PMID- 19296093 TI - Identification of beta-carotene in an evaporitic matrix--evaluation of Raman spectroscopic analysis for astrobiological research on Mars. AB - Since evaporitic rocks on the Martian surface could (or still can) serve as potential habitats for microbial life on Mars, there is a reasonable possibility that these rocks may sustain molecular remnants as evidence for the presence of extinct or extant living organisms on Mars and that beta-carotene could be a suitable biomarker. In this paper, Raman microspectrometry was tested as a nondestructive method of determining the lowest detectable beta-carotene content in experimentally prepared evaporitic matrices--namely, gypsum, halite and epsomite. Two excitation wavelengths were compared--514.5 nm, because of the resonance Raman enhancement in the carotenoid analysis, and 785 nm, as a more universal wavelength now much used in the detection of biomolecules terrestrially. Mixtures were measured directly as well as with a laser beam penetrating the crystals of gypsum and epsomite. We have obtained beta-carotene signals at the 0.1 to 10 mg kg(-1) level--the number of registered beta-carotene Raman bands differed depending on the particular mineral matrix and the excitation wavelength. Concentrations of beta-carotene of about one order of magnitude higher were identified when analysed through single crystals of gypsum and epsomite, respectively. PMID- 19296094 TI - Neural correlates of audiovisual motion capture. AB - Visual motion can affect the perceived direction of auditory motion (i.e., audiovisual motion capture). It is debated, though, whether this effect occurs at perceptual or decisional stages. Here, we examined the neural consequences of audiovisual motion capture using the mismatch negativity (MMN), an event-related brain potential reflecting pre-attentive auditory deviance detection. In an auditory-only condition occasional changes in the direction of a moving sound (deviant) elicited an MMN starting around 150 ms. In an audiovisual condition, auditory standards and deviants were synchronized with a visual stimulus that moved in the same direction as the auditory standards. These audiovisual deviants did not evoke an MMN, indicating that visual motion reduced the perceptual difference between sound motion of standards and deviants. The inhibition of the MMN by visual motion provides evidence that auditory and visual motion signals are integrated at early sensory processing stages. PMID- 19296095 TI - The Open AUC Project. AB - Progress in analytical ultracentrifugation (AUC) has been hindered by obstructions to hardware innovation and by software incompatibility. In this paper, we announce and outline the Open AUC Project. The goals of the Open AUC Project are to stimulate AUC innovation by improving instrumentation, detectors, acquisition and analysis software, and collaborative tools. These improvements are needed for the next generation of AUC-based research. The Open AUC Project combines on-going work from several different groups. A new base instrument is described, one that is designed from the ground up to be an analytical ultracentrifuge. This machine offers an open architecture, hardware standards, and application programming interfaces for detector developers. All software will use the GNU Public License to assure that intellectual property is available in open source format. The Open AUC strategy facilitates collaborations, encourages sharing, and eliminates the chronic impediments that have plagued AUC innovation for the last 20 years. This ultracentrifuge will be equipped with multiple and interchangeable optical tracks so that state-of-the-art electronics and improved detectors will be available for a variety of optical systems. The instrument will be complemented by a new rotor, enhanced data acquisition and analysis software, as well as collaboration software. Described here are the instrument, the modular software components, and a standardized database that will encourage and ease integration of data analysis and interpretation software. PMID- 19296096 TI - Toward a mechanical control of drug delivery. On the relationship between Lipinski's 2nd rule and cytosolic pH changes in doxorubicin resistance levels in cancer cells: a comparison to published data. AB - Based on molecular and physiological resemblance, the mechanism that controls drug bioavailability and toxicity also shares strong similarities to the one that controls drug resistance. In both cases, this mechanism relies on the expression of drug transporters and the physico-chemical properties of drugs, which together alter the intracellular accumulation of chemicals in cells or tissues. However, a parameter that is central and has received great attention in the field of bioavailability, but almost none in the field of drug resistance, is the molecular weight of drugs. In the former area, it is well known that to achieve a reasonable bioavailability, drugs must have-among other properties-a molecular weight less than 500, known as Lipinski's 2nd rule. Accordingly, it is worth questioning whether a similar rule exists in the field of drug resistance and what subsequent mechanism would control the membrane permeability to drugs as a function of their molecular weight. I demonstrate here that cytosolic pH fixes the molecular weight of drugs entering cells, by altering the cell membrane mechanical properties and that, both cytosolic pH and membrane mechanical properties are needed and sufficient to explain doxorubicin resistance levels in different cancerous cell lines. Finally, I discuss the efficiency of a drug handling activity by transporters in MDR and suggest ways to control drug delivery mechanically. In addition, and for the first time, the literal expression of a Law similar to Lipinski's 2nd rule will be described as a function of cytosolic pH and lipid number asymmetry. PMID- 19296097 TI - Production of a novel glycolipid biosurfactant, mannosylmannitol lipid, by Pseudozyma parantarctica and its interfacial properties. AB - The development of a novel glycolipid biosurfactant was undertaken using the high level producers of mannosylerythritol lipids (MELs) such as Pseudozyma parantarctica, Pseudozyma antarctica, and Pseudozyma rugulosa. Besides the conventional MELs (MEL-A, MEL-B, and MEL-C), these yeasts produced an unknown glycolipid when they were cultivated in a medium containing 4% (w/v) olive oil and 4% (w/w) mannitol as the carbon source. The unknown glycolipid extracted from the culture medium of P. parantarctica JCM 11752(T) displayed the spot with lower mobility than that of known MELs on TLC and provided mainly two peaks identical to mannose and mannitol on high-performance liquid chromatography after acid hydrolysis. Based on structural analysis by (1)H and (13)C nuclear magnetic resonance, the novel glycolipid was composed of mannose and mannitol as the hydrophilic sugar moiety and was identified as mannosylmannitol lipid (MML). Of the strains tested, P. parantarctica JCM 11752(T) gave the best yield of MML (18.2 g/L), which comprised approximately 35% of all glycolipids produced. We further investigated the interfacial properties of the MML, considering the unique hydrophilic structure. The observed critical micelle concentration (CMC) and the surface tension at CMC of the MML were 2.6 x 10(-6) M and 24.2 mN/m, respectively. In addition, on a water-penetration scan, the MML efficiently formed not only the lamella phase (Lalpha) but also the myelins at a wide range of concentrations, indicating its excellent self-assembling properties and high hydrophilicity. The present glycolipid should thus facilitate the application of biosurfactants as new functional materials. PMID- 19296098 TI - Effect of ultrafine-grained titanium surfaces on adhesion of bacteria. AB - The influence of the ultrafine crystallinity of commercial purity grade 2 (as received) titanium and titanium modified by equal channel angular pressing (modified titanium) on bacterial attachment was studied. A topographic profile analysis of the surface of the modified titanium revealed a complex morphology of the surface. Its prominent micro- and nano-scale features were 100-200-nm-scale undulations with 10-15 microm spacing. The undulating surfaces were nano-smooth, with height variations not exceeding 5-10 nm. These surface topography characteristics were distinctly different from those of the as-received samples, where broad valleys (up to 40-60 microm) were detected, whose inner surfaces exhibited asperities approximately 100 nm in height spaced at 1-2 microm. It was found that each of the three bacteria strains used in this study as adsorbates, viz. Staphylococcus aureus CIP 68.5, Pseudomonas aeruginosa ATCC 9025 and Escherichia coli K12, responded differently to the two types of titanium surfaces. Extreme grain refinement by ECAP resulted in substantially increased numbers of cells attached to the surface compared to as-received titanium. This enhanced degree of attachment was accompanied with an increased level of extracellular polymeric substances (EPS) production by the bacteria. PMID- 19296099 TI - Biological analysis of the deletion mutants of Staphylococcal enterotoxin C2. AB - To investigate the functional domains involved in the biological activity of staphylococcal enterotoxin (SEC2), a series of SEC2 mutants were constructed. Deletion of the last 77 amino acids at the C-terminus of SEC2 did not affect its native superantigen and fever activities, and further removal of the C-terminal residues reduced SEC2 activities significantly. On the other hand, the mutants lacking 18 or more N-terminal residues severely impaired superantigen activity. These data indicated that the functional regions for the biological activities of SEC2 were confined to N-terminal domain, further implied that the proper three dimensional structure of SEC2 is not needed for its biological activities. Our results deliver valuable information that it is possible to design new SEC2 immunotherapeutic agents which have the superantigen activity and low molecular weight for permeability. PMID- 19296100 TI - Initial results of in vivo high-resolution morphological and biochemical cartilage imaging of patients after matrix-associated autologous chondrocyte transplantation (MACT) of the ankle. AB - OBJECTIVE: The aim of this study was to use morphological as well as biochemical (T2 and T2* relaxation times and diffusion-weighted imaging (DWI)) magnetic resonance imaging (MRI) for the evaluation of healthy cartilage and cartilage repair tissue after matrix-associated autologous chondrocyte transplantation (MACT) of the ankle joint. MATERIALS AND METHODS: Ten healthy volunteers (mean age, 32.4 years) and 12 patients who underwent MACT of the ankle joint (mean age, 32.8 years) were included. In order to evaluate possible maturation effects, patients were separated into short-term (6-13 months) and long-term (20-54 months) follow-up cohorts. MRI was performed on a 3.0-T magnetic resonance (MR) scanner using a new dedicated eight-channel foot-and-ankle coil. Using high resolution morphological MRI, the magnetic resonance observation of cartilage repair tissue (MOCART) score was assessed. For biochemical MRI, T2 mapping, T2* mapping, and DWI were obtained. Region-of-interest analysis was performed within native cartilage of the volunteers and control cartilage as well as cartilage repair tissue in the patients subsequent to MACT. RESULTS: The overall MOCART score in patients after MACT was 73.8. T2 relaxation times (approximately 50 ms), T2* relaxation times (approximately 16 ms), and the diffusion constant for DWI (approximately 1.3) were comparable for the healthy volunteers and the control cartilage in the patients after MACT. The cartilage repair tissue showed no significant difference in T2 and T2* relaxation times (p > or = 0.05) compared to the control cartilage; however, a significantly higher diffusivity (approximately 1.5; p < 0.05) was noted in the cartilage repair tissue. CONCLUSION: The obtained results suggest that besides morphological MRI and biochemical MR techniques, such as T2 and T2* mapping, DWI may also deliver additional information about the ultrastructure of cartilage and cartilage repair tissue in the ankle joint using high-field MRI, a dedicated multichannel coil, and sophisticated sequences. PMID- 19296101 TI - Pilomatricomas: the diagnostic value of ultrasound. AB - OBJECTIVE: The purpose of this study was to analyze statistically significant diagnostic factors for pilomatricoma on the basis of ultrasonographic features. METHODS: Sonographic images were retrospectively reviewed from 44 pilomatricomas, and from 43 control subjects with other subcutaneous tumors. Two radiologists determined the tumoral shape, margin, echotexture, echogenicity, posterior shadowing, posterior enhancement, hypoechoic rim, internal calcification, and vascularity. RESULTS: The reliable diagnostic factors for pilomatricoma were hypoechogenicity (P < 0.001), heterogenicity (P < 0.05), internal calcification (P < 0.001), hypoechoic rim (P < 0.001), and posterior shadowing (P < 0.001). Scattered dots were the most common patterns of internal calcification. A combination of hypoechogenicity, heterogenicity, internal calcification of scattered-dot pattern, and a hypoechoic rim was a statistically significant difference between the two groups (P < 0.001; odds ratio, 21). CONCLUSIONS: The features of heterogeneous echotexture, internal echogenic foci in scattered-dot pattern, and a hypoechoic rim or posterior shadowing itself could be discriminative ultrasonographic criteria for differentiating pilomatricomas from other subcutaneous tumors. PMID- 19296102 TI - Noninvasive imaging of tumor integrin expression using (18)F-labeled RGD dimer peptide with PEG (4) linkers. AB - PURPOSE: Various radiolabeled Arg-Gly-Asp (RGD) peptides have been previously investigated for tumor integrin alpha(v)beta(3) imaging. To further develop RGD radiotracers with enhanced tumor-targeting efficacy and improved in vivo pharmacokinetics, we designed a new RGD homodimeric peptide with two PEG(4) spacers (PEG(4) = 15-amino-4,7,10,13-tetraoxapentadecanoic acid) between the two monomeric RGD motifs and one PEG(4) linker on the glutamate alpha-amino group ((18)F-labeled PEG(4)-E[PEG(4)-c(RGDfK)](2), P-PRGD2), as a promising agent for noninvasive imaging of integrin expression in mouse models. METHODS: P-PRGD2 was labeled with (18)F via 4-nitrophenyl 2-(18)F-fluoropropionate ((18)F-FP) prosthetic group. In vitro and in vivo characteristics of the new dimeric RGD peptide tracer (18)F-FP-P-PRGD2 were investigated and compared with those of (18)F-FP-P-RGD2 ((18)F-labeled RGD dimer without two PEG(4) spacers between the two RGD motifs). The ability of (18)F-FP-P-PRGD2 to image tumor vascular integrin expression was evaluated in a 4T1 murine breast tumor model. RESULTS: With the insertion of two PEG(4) spacers between the two RGD motifs, (18)F-FP-P-PRGD2 showed enhanced integrin alpha(v)beta(3)-binding affinity, increased tumor uptake and tumor-to-nontumor background ratios compared with (18)F-FP-P-RGD2 in U87MG tumors. MicroPET imaging with (18)F-FP-P-PRGD2 revealed high tumor contrast and low background in tumor-bearing nude mice. Biodistribution studies confirmed the in vivo integrin alpha(v)beta(3)-binding specificity of (18)F-FP-P-RGD2. (18)F-FP P-PRGD2 can specifically image integrin alpha(v)beta(3) on the activated endothelial cells of tumor neovasculature. CONCLUSION: (18)F-FP-P-PRGD2 can provide important information on integrin expression on the tumor vasculature. The high integrin binding affinity and specificity, excellent pharmacokinetic properties and metabolic stability make the new RGD dimeric tracer (18)F-FP-P PRGD2 a promising agent for PET imaging of tumor angiogenesis and for monitoring the efficacy of antiangiogenic treatment. PMID- 19296103 TI - Prognostic significance of stress myocardial ECG-gated perfusion imaging in asymptomatic patients with diabetic chronic kidney disease on initiation of haemodialysis. AB - PURPOSE: Diabetic patients with chronic kidney disease (CKD) frequently develop cardiac events within several years of the initiation of haemodialysis. The present study assesses the prognostic significance of stress myocardial ECG-gated perfusion imaging (MPI) in patients with diabetic CKD requiring haemodialysis. METHODS: Fifty-five asymptomatic patients with diabetic stage V CKD and no history of heart disease scheduled to start haemodialysis were enrolled in this study (56 +/- 11 years old; 49 with type 2 diabetes mellitus). All patients underwent (201)Tl stress ECG-gated MPI 1 month before or after the initiation of haemodialysis to assess myocardial involvement. We evaluated SPECT images using 17-segment defect scores graded on a 5-point scale, summed stress score (SSS) and summed difference scores (SDS). The patients were followed up for at least 2 years (42 +/- 15 months) to determine coronary intervention (CI) and heart failure (HF) as soft events and acute myocardial infarction (AMI) and all causes of deaths as hard events. RESULTS: The frequencies of myocardial ischaemia, resting perfusion defects, low ejection fraction and left ventricular (LV) dilatation were 24, 20, 29 and 49%, respectively. Ten events (18%) developed during the follow-up period including four CI, one HF, one AMI and four sudden deaths. Multivariate Cox analysis selected SDS (p = 0.0011) and haemoglobin A(1c) (HbA(1c)) (p = 0.0076) as independent prognostic indicators for all events. CONCLUSION: Myocardial ischaemia, in addition to glycaemic control, is a strong prognostic marker for asymptomatic patients with diabetic CKD who are scheduled to start haemodialysis. Stress MPI is highly recommended for the management and therapeutic stratification of such patients. PMID- 19296104 TI - Symmetric giant xanthogranulomas in Erdheim-Chester disease. PMID- 19296105 TI - Demonstration of metastatic tumour growth following vessel structures by PET/CT. PMID- 19296106 TI - Disseminated tuberculosis infection: a 'super' 18F-FDG PET/CT appearance. PMID- 19296107 TI - Dissociation between myocardial perfusion scintigraphy and angiographic findings explained by intravascular ultrasonography. PMID- 19296108 TI - Rapid assessment of rest myocardial ischaemia in the Chest Pain Unit. PMID- 19296109 TI - Comparison of various surgical methods in the treatment of implant-related infection. AB - The treatment of implant-related infections is troublesome. This study was conducted to compare the effectiveness of three different surgical modalities in the treatment of implant-related infection. A total of 32 Wistar albino rats were randomised into four groups after the establishment of implant-related infection: no treatment, surgical debridement, antibiotic-loaded bone cement and antibiotic loaded autogenous bone. Microbiological colony counts were made at the sixth week in order to evaluate the effectiveness of of the treatments. The antibiotic loaded bone cement group revealed superior results compared with the other groups in terms of reduction of microbiological colonies. Three animals in the bone cement group revealed extensive infection. Although antibiotic-loaded bone cement showed superiority over other treatment modalities, it should be employed after an unsuccessful trial of debridement because of the risk of extensive infection. PMID- 19296110 TI - Different clinical importance of FLT3 internal tandem duplications in AML according to FAB classification: possible existence of distinct leukemogenesis involving monocyte differentiation pathway. AB - Impact of FLT3 receptor tyrosine kinase activation via internal tandem duplication (ITD) of the juxtamembrane region on outcome of acute myeloid leukemia (AML) is still controversial. Recent researches reveal a role of FLT3 in monocyte differentiation in hematopoiesis. We analyzed the clinical impact of FLT3 alterations in adult AML patients excluding acute promyelocytic leukemia (APL) who received induction chemotherapy according to morphologic classification. Retrospective review of medical records from three centers in Korea between 1997 and 2007 was performed. Polymerase chain reaction was performed on genomic DNA derived from blood samples of patients before induction chemotherapy for FLT3-ITD detection. We assessed overall survival (OS), first disease-free survival (1-DFS), and response to induction chemotherapy. One hundred eighty-four patients (median age 49.1 years, range 16.0-76.5) with AML excluding APL received induction chemotherapy from three centers. FLT3-ITD was detected in 22 patients. One hundred forty-one patients were below age 60. One hundred seventy-nine patients received induction chemotherapy with cytarabine and idarubicin (AId) regimen. One hundred nineteen patients achieved complete remission (CR) after first induction chemotherapy. FLT3-ITD was not related to achievement of CR. 1-DFS was longer in patients without FLT3-ITD (median 1-DFS 16.5 vs. 8.5 months, p = 0.025). 1-DFS was not different according to FLT3-ITD status in nonmonocyte lineage leukemia (p = 0.355), while 1-DFS was shorter in monocyte lineage leukemia for FLT3-ITD positive patients (20.9 vs. 2.4 months, p < 0.001). FLT3-ITD had no impact on OS except for monocyte lineage, where OS was significantly shorter in FLT3-ITD positive group (39.4 vs. 6.0 months, p = 0.026). Moreover FLT3-ITD was stronger prognostic factors in monocyte lineage AML than risk stratification based on cytogenetics. Status of FLT3-ITD should be analyzed differently in AML patients according to morphologic profile. FLT3-ITD is a predictive and prognostic marker only in monocyte lineage patients. This result suggests an existence of distinct subset of monocyte lineage AML with leukemogenesis involving FLT3 activating pathway. PMID- 19296111 TI - [Amyloidoses in the Berlin Museum of Medical History of the Charite]. AB - The Berliner Medizinhistorische Museum (Berlin Museum of Medical History) of the Charite is located in its own separate building and was officially opened in 1899. It currently houses 10,000 specimens, of which 23 were labelled with the diagnosis "amyloid" or "amyloidosis". In this retrospective study we aimed to histologically verify the diagnosis, classify the amyloid deposits immunohistochemically and correlate the type of amyloid with clinico-pathological data. The specimens were obtained between 1866 and 1987 and included 17 kidneys, five spleens and one liver. The diagnosis could be confirmed histologically using Congo red staining and polarization microscopy in 22 specimens. However, the diagnosis could not be confirmed in the oldest specimen, which had been labelled by Rudolf Virchow himself. Immunohistochemically amyloid was classified as either AA amyloidosis (19 cases) or AL amyloidosis (two cases). Tuberculosis was the most common cause of AA amyloidosis. This study shows that a surgical pathological re-evaluation of historical specimens can verify the original diagnosis. This is historically fascinating and also offers a valuable addition to student teaching. PMID- 19296112 TI - Evolution of carbonic anhydrases in fungi. AB - The ubiquitous metalloenzyme carbonic anhydrase (CA) catalyzes the interconversion of carbon dioxide and bicarbonate. This enzyme has been investigated in mammals, plants, algae, bacteria, archaea and fungi. Based on distinct structural characteristics, CAs can be assigned to five independently evolved classes (alpha, beta, gamma, delta and zeta). beta-CAs can be further subdivided into plant-type and cab-type sub-classes. The recent characterization of CAs in fungi led us to initiate a systematic search for these enzymes in filamentous ascomycetes. The genomes of basidiomycetes and hemiascomycetous yeasts contain only beta-CAs, while the filamentous ascomycetes also possess genes encoding alpha-class CAs. Here, we present a phylogenetic analysis of 97 fungal CA sequences that addresses the diversification of fungal CAs. During evolution various gene duplication and gene loss events seem to be the cause for the multiplicity of CAs in filamentous ascomycetes. Our data revealed that during the evolution of filamentous ascomycetes, a gene encoding the plant-type beta-CA was duplicated, resulting in two closely related isoforms, one with and one without an N-terminal mitochondrial target sequence (MTS). The acquisition of the MTS most likely took place after the gene duplication event and after the evolutionary separation of the fungal orders Sordariales and Eurotiales. PMID- 19296113 TI - Long-term safety and outcome of a temporary self-expanding metallic stent for achalasia: a prospective study with a 13-year single-center experience. AB - To prospectively evaluate the long-term clinical safety and efficacy of a newly designed self-expanding metallic stent (SEMS) in the treatment of patients with achalasia. Seventy-five patients with achalasia were treated with a temporary SEMS with a 30-mm diameter. The SEMSs were placed under fluoroscopic guidance and removed by gastroscopy 4-5 days after stent placement. Follow-up data focused on dysphagia score, technique and clinical success, clinical remissions and failures, and complications and was performed at 6 months, 1 year, and within 3 to 5 years, 5 to 8 years, 8 to 10 years, and >10 years postoperatively. Stent placement was technically successful in all patients. Complications included stent migration (n = 4, 5.33%), chest pain (n = 28, 38.7%), reflux (n = 15, 20%), and bleeding (n = 9, 12%). No perforation or 30-day mortality occurred. Clinical success was achieved in all patients 1 month after stent removal. The overall remission rates at 6 months, 1, 1-3, 3-5, 5-8, 8-10, and >10 year follow-up periods were 100%, 96%, 93.9%, 90.9%, 100%, 100%, and 83.3%, respectively. Stent treatment failed in six patients, and the overall remission rate in our series was 92%. The median and mean primary patencies were 2.8 +/- 0.28 years (95% CI: 2.25-3.35) and 4.28 +/- 0.40 years (95% CI: 3.51-5.05), respectively. The use of temporary SEMSs with 30-mm diameter proved to be a safe and effective approach for managing achalasia with a long-term satisfactory clinical remission rate. PMID- 19296114 TI - Pseudotumor cerebri: as a cause for early deterioration after Chiari I malformation surgery. AB - INTRODUCTION: Chiari I malformation is associated with a small posterior fossa which cannot accommodate a growing hindbrain. Pseudotumor cerebri has been linked to developmental posterior fossa malformations. CASE REPORTS: The authors present two cases of early-deterioration post-Chiari I malformation surgery in a young and an adult patient, which were linked to raised intracranial pressure and had a stormy post-operative period. The pathophysiology of pseudotumor cerebri in early post-operative complications of Chiari malformation surgery is addressed along with arguments favoring its association. DISCUSSION: Potential clinical risk factors and red flags linking pseudotumor cerebri and Chiari malformation patients are discussed. The management of post-operative complications produced by the association is addressed. PMID- 19296115 TI - Endoscopic management of brainstem injury due to ventriculoperitoneal shunt placement. AB - INTRODUCTION: We report a patient who suffered from brainstem injury following ventriculoperitoneal (VP) shunt placement in the fourth ventricle. DISCUSSION: A 20-year-old man with complex hydrocephalus and trapped fourth ventricle underwent a suboccipital placement of a VP shunt. Postprocedure patient developed double vision. Magnetic resonance imaging showed that the catheter was penetrating the dorsal brainstem at the level of the pontomedullary junction. Patient was referred to our Neuroendoscopic Clinic. Physical exam demonstrated pure right VI cranial nerve palsy. Patient underwent flexible endoscopic exploration of the ventricular system. Some of the endoscopic findings were severe aqueductal stenosis and brainstem injury from the catheter. Aqueductoplasty, transaqueductal approach into the fourth ventricle, and endoscopic repositioning of the catheter were some of the procedures performed. Patient recovered full neurological function. The combination of endoscopic exploration and shunt is a good alternative for patients with complex hydrocephalus. A transaqueductal approach to the fourth ventricle with flexible scope is an alternative for fourth ventricle pathology. PMID- 19296116 TI - Use of 11C-methionine positron emission tomography in basal germinoma: assessment of treatment response and residual tumor. AB - INTRODUCTION: Prognosis of germinoma arising in the basal ganglia or thalamus is worse compared to that in the pineal or suprasellar region. One of the reasons for poor prognosis is the difficulty in evaluating the efficacy of treatment by conventional neuroimaging tools. PET STUDIES: The usefulness of (11)C-methionine (MET) positron emission tomography (PET) in monitoring the biological nature of brain tumors has been proved in glioma patients. CASE REPORTS: Herein, we describe MET-PET findings in three cases of germinomas in the basal ganglia or thalamus and discuss the use of MET-PET in the assessment of treatment response and residual tumor for the next treatment strategy. The patients showed transient increase of MET uptake in the lesions after the initial treatment. Although we did not perform histological verification, MET- PET findings suggested that active tumor cells were still alive in the lesions after the initial treatment. MET uptake gradually decreased during the course of intensive therapy in these patients. MET-PET also revealed germinoma invasion in the brain before discernible signal abnormality or mass lesion in conventional magnetic resonance images in two patients. DISCUSSION: Further studies including histological verification and long-term follow-up might validate the use of MET-PET in monitoring the treatment efficacy and evaluation of active residual tumor after the treatment. CONCLUSION: Until we understand what MET uptake truly represents, treatment strategy based on MET uptake must be carefully designed to prevent overtreatment and resultant complications. PMID- 19296117 TI - Colorectal cancer screening in Europe: differences in approach; similar barriers to overcome. AB - Worldwide diagnoses of bowel cancer approximate an estimated one million new cases per year, comprising 9% of all cancer cases, and this has continued to increase over the last 25 years. With the association between cancer risk and increasing age, together with the suggestion that by 2015 there will be a 22% increase in the proportion of the population aged over 65 years and a 50% increase in the proportion of people aged over 80 years, there is likely to be a significant increase in the demand on cancer services throughout Europe and the rest of the world. This article discusses the current state of bowel cancer screening within Europe. PMID- 19296118 TI - A COX-2 inhibitor combined with chemoradiation of locally advanced rectal cancer. PMID- 19296119 TI - Detection of laryngeal tumors and tumoral extension by multislice computed tomography-virtual laryngoscopy (MSCT-VL). AB - Virtual endoscopy is becoming a widely used non-invasive clinical diagnostic tool. The present study was designed to compare the sensitivity and specificity of the conventional endoscopy and virtual laryngoscopy with respect to laryngeal masses. A total of 38 patients (20 males, 18 females, mean age 61 years) with the complaint of hoarseness were included in the study. Laryngeal mucosa, lumen and mass pathology were evaluated initially by direct endoscopy and then by virtual laryngoscopy during multislice CT of the larynx. Histopathologic evaluation of the masses was also made. The main pathology of the patients was found to be laryngeal masses (60% of patients, one mass for each patient), which were polyps (n = 8), papilloma (n = 4) and carcinoma (n = 11) according to histopathologic evaluation. Retrospective evaluation of 6 lesions detected in virtual but not in conventional laryngoscopy resulted with the finding of viscous-dense mucous secretion. On the contrary, three lesions detected by conventional laryngoscopy could not be detected by virtual evaluation. A total of six patients were evaluated and considered as normal both by conventional and virtual laryngoscopic examinations. Sensitivity of the virtual laryngoscopy was 88% (23/26) while its specificity was only 50% (6/12). Positive and negative predictive values were 79% (23/29) and 66% (6/9), respectively. Accuracy of the virtual laryngoscopy was 76% (29/38). Virtual laryngoscopy is not an alternative to conventional laryngoscopy but may assist direct endoscopy without causing additional radiation exposure or discomfort to the patient. The three-dimensional contribution to interpretation of the results and subsequent manipulation of the data can be used for educational and surgical purposes. PMID- 19296120 TI - Chromatin dynamics is correlated with replication timing. AB - Discrete chromatin domains (ChrD), containing an average of approximately 1 Mbp DNA, represent the basic structural units for the regulation of DNA organization and replication in situ. In this study, a bio-computational approach is employed to simultaneously measure the translational motion of large populations of ChrD in the cell nucleus of living cells. Both movement and configurational changes are strikingly higher in early S-phase replicating ChrD compared to those that replicate in mid and late S-phase. The chromatin dynamics was not sensitive to transcription inhibition by alpha-amanitin but was significantly reduced by actinomycin D treatment. Since a majority of active genes replicate in early S phase, our results suggest a correlation between levels of chromatin dynamics and chromatin poised for active transcription. Analysis of ChrD colocalization with transcription sites and cDNA with ChrD and transcription sites further supports this proposal. PMID- 19296121 TI - The Mi-2/NuRD complex associates with pericentromeric heterochromatin during S phase in rapidly proliferating lymphoid cells. AB - Chromosomal replication results in the duplication not only of DNA sequence but also of the patterns of histone modification, DNA methylation, and nucleoprotein structure that constitute epigenetic information. Pericentromeric heterochromatin in human cells is characterized by unique patterns of histone and DNA modification. Here, we describe association of the Mi-2/NuRD complex with specific segments of pericentromeric heterochromatin consisting of Satellite II/III DNA located on human chromosomes 1, 9, and 16 in some but not all cell types. This association is linked in part to DNA replication and chromatin assembly and may suggest a role in these processes. Mi-2/NuRD accumulation is independent of Polycomb association and is characterized by a unique pattern of histone modification. We propose that Mi-2/NuRD constitutes an enzymatic component of a pathway for assembly and maturation of chromatin utilized by rapidly proliferating lymphoid cells for replication of constitutive heterochromatin. PMID- 19296122 TI - Evaluation of potential retinal toxicity of adalimumab (Humira). AB - PURPOSE: The purpose of this study is to evaluate the retinal toxicity of two doses of adalimumab (Humira), a recombinant human IgG1 monoclonal antibody specific for human tumor necrosis factor (TNF), when injected intravitreally in rabbits. METHODS: Sixteen male pigmented rabbits (divided into two groups, eight animals per group) were used for this study. Two concentrations of adalimumab were tested: 0.5 mg/0.1 ml and 5 mg/0.1 ml. Each concentration was injected intravitreally randomly in one eye (study group) of each rabbit (group I received 0.5 mg/0.1 ml and group II received 5.0 mg/0.1 ml), while in the other eye (control group) 0.1 ml of sterile balanced saline solution (BSS) was injected. Slit-lamp and funduscopic examinations were performed every second day for 2 weeks for signs of infection, inflammation and toxicity. A baseline electroretinogram (ERG) was performed before the experiment and at the last follow-up day (day 14). ERG examination followed ISCEV standards. At the last follow-up day, the animals were sacrificed and the enucleated eyes were prepared for histological evaluation of retinal toxicity. RESULTS: No differences in ERG responses at photopic and scotopic conditions were observed in eyes injected with either concentration of adalimumab or BSS. Furthermore, histologic examination of the retina in the enucleated eyes (in all groups) did not demonstrate any evidence of drug toxicity. CONCLUSIONS: Intravitreal adalimumab did not appear toxic to the retina in this experimental model at concentrations of 0.5 and 5 mg. If found safe in additional studies, intravitreally injected adalimumab could be evaluated for efficacy in the treatment of inflammatory eye conditions. PMID- 19296123 TI - The false-negative rate of fine-needle aspiration cytology for diagnosing thyroid carcinoma in thyroid nodules. AB - PURPOSE: Our aim was to define the false-negative rate of fine-needle aspiration cytology (FNAC) for diagnosing thyroid carcinoma in thyroid nodules <4 cm versus > or =4 cm. MATERIALS AND METHODS: Six hundred sixty-two patients with thyroid nodules who underwent FNAC and surgery at our institution were analyzed. The association of predictive factors with thyroid carcinoma was evaluated. The sensitivity, specificity, and the false-negative rate of FNAC were calculated. RESULTS: The incidence of thyroid carcinoma was significantly higher in nodules > or =4 cm (24%) compared with nodules <4 cm (12%). The false-negative rates of FNAC were 2% in all nodules and 1.3% and 4.3% in nodules <4 cm and > or =4 cm (p = 0.9), respectively. Sensitivity and specificity of FNAC were 90% and 79%, respectively. CONCLUSIONS: The false-negative rate of FNAC is low for thyroid nodules <4 cm and for nodules > or =4 cm. PMID- 19296124 TI - In vitro modulation of the cardiac ryanodine receptor activity by Homer1. AB - The Homer protein family allows clustering and/or functional modulation of many proteins from different calcium signalling complexes including those formed by the ryanodine receptor (RyR) Ca(2+) release channel in skeletal muscle and the heart. Homer1b/c and the cardiac RyR (RyR2) are strongly expressed in the heart and neurons where their interaction with each other may modulate Ca(2+) signalling. However, functional interactions between Homer1b and RyR2 have been poorly defined. Our preliminary data and similar consensus binding sites for Homer in RyR2 and skeletal RyR (RyR1) proteins, led to the hypothesis that Homer may similarly regulate both RyR isoforms. Single-channel and [(3)H]ryanodine binding data showed that RyR2 and RyR1 activity increased to a maximum with ~50 100 nM Homer1b and fell with Homer1b > 200 nM. Homer1b (50 nM) activated RyR2 and RyR1 at all cytosolic [Ca(2+)]; estimated EC(50) value of RyR2 diminished from ~2.8 microM Ca(2+) (control) to ~1.9 microM Ca(2+) in the presence of 50 nM Homer1b. Short Homer1 (lacking the coiled-coil multimerisation domain) and Homer1b similarly modulated RyR2, indicating an action through ligand binding, not mutimerisation. These actions of Homer were generally similar in RyR2 and RyR1. The strong functional interactions suggest that Homer1 is likely to be an endogenous modulator of RyR channels in the heart and neurons as well as in skeletal muscle. PMID- 19296125 TI - A lentivirally delivered photoactivatable GFP to assess continuity in the endoplasmic reticulum of neurones and glia. AB - The endoplasmic reticulum (ER) is the largest intracellular membranous organelle. Functions of the ER are many and diverse, which include various biosynthetic, transport and signalling roles, central to cellular physiology, such as the biosynthesis of membrane and secretory proteins and the regulation of intracellular calcium. Its continuous lumen also serves as a highway for the distribution of proteins and ions to different regions of the cell, independent of the cytosol. The ER is an excitable organelle, capable of generating a regenerative wave of calcium release, which can propagate along the endomembrane throughout the entire cell, serving as a system of intracelluar communication in polarised cells. Nowhere is this feature of ER function more crucial than in neurones. The extremely polarised nature of nerve cells presents a unique challenge for the global co-ordination of localised physiological events such as growth cone guidance and synaptic plasticity. Clearly, the physical continuity of the neuronal ER lumen is central to its functionality as a conduit for communication. To further probe the continuity of ER in neurones and glia, we developed LV-PA-pIN-KDEL, a photoactivatable analogue of our recently described vector LV-pIN-KDEL, a lentivirally delivered ER-targeting soluble GFP. We demonstrate the ability of this vector to transduce astrocytes and neurones in culture and in cortical explants. Furthermore, we exploit the photoactivatable attributes of the vector together with a focal laser photoactivation protocol to reveal the continuous nature of the ER lumen in these cell types, presenting the first direct evidence of an astrocytic ER luminal continuum and providing more data to support the existence of a single ER lumen in neurones. PMID- 19296126 TI - Evidence for programmed cell death and activation of specific caspase-like enzymes in the tomato fruit heat stress response. AB - The tomato (Lycopersicon esculentum) fruit is the best available model to study the stress response of fleshy fruit. Programmed cell death (PCD) plays an important role in stress responses in mammals and plants. In this study, we provide evidence that PCD is triggered in the tomato fruit heat stress response by detection of the sequential diagnostic PCD events, including release of cytochrome c, activation of caspase-like proteases and the presence of TUNEL positive nuclei. Investigating the time course of these events for 12 h after heat treatment indicated that cytochrome c release and caspase-like protease activation occurred rapidly and were consistent with the onset of DNA fragmentation. In addition, LEHDase and DEVDase enzymes were specifically activated in tomato fruit pericarp during the heat treatment and recovery time. There was no significant activation of YVADase or IETDase proteases. Preincubation of pericarp discs with the broad-spectrum, cell-permeable caspase inhibitor Z-VAD-FMK, suppressed heat-induced cell death measured by trypan blue, accompanied by a decrease in LEHDase and DEVDase activities. PMID- 19296127 TI - Disturbance of circadian gene expression in breast cancer. AB - To explore the mechanism of the disruption of circadian rhythm in breast cancer, we examined the expression of nine circadian genes in 53 newly diagnosed breast cancers by immunohistochemical staining, mutational analysis, and methylation analysis of the promoter of circadian genes. Our results showed that 37 of the 53 breast cancer tissues had hypermethylation on the promoters of PER1, PER2, CRY1, or BMAL1. Twenty-five out of 53 paired noncancerous (normal) tissues had methylation on the promoter of PER1 or CRY1. Our results indicated a higher frequency of concurrent methylation of PER1 and CRY1 promoters in cancerous and normal tissues. Promoter methylation of the PER1 correlates with c-erbB2 immunohistochemical reaction of > or = 2+ (p = 0.012) and has a strong inverse correlation with estrogen receptor positivity (p = 0.016). We further analyzed the patterns of circadian gene expression by immunohistochemical methods and found that homogeneous expression of PER2 or BMAL1 is significantly associated with lymph node metastasis and poor prognosis. PER2 heterogeneous expression correlates with <2+ c-erbB2 immunohistochemical reaction. Heterogeneous expression of CLOCK is associated significantly with 3-year survival. In conclusion, the expression pattern of circadian genes might be a biomarker for the prognosis of breast cancer. PMID- 19296128 TI - Semaphorin3A immunohistochemical expression in human meningiomas: correlation with the microvessel density. AB - The immunoexpression of the antiangiogenic factor semaphorin3A (SEMA3A) was evaluated in a series of meningiomas. Then, its correlations with the microvessel density (MVD) of the tumors and with the clinicopathological parameters as well with the survival time or recurrence-free interval were investigated. A positive SEMA3A immunostaining was found in most of meningiomas and a significant association was found between a high expression of this protein and a low MVD of the tumors. Moreover, a low SEMA3A immunoexpression was significantly correlated with a higher recurrence rate of meningiomas. In conclusion, our findings suggest a role for SEMA3A as an antiangiogenic factor in meningiomas with its decrease being associated with the development of recurrences. The supplementation of SEMA3A might be used in novel therapeutic antiangiogenic strategies to prevent the recurrence of highly vascularized meningiomas. PMID- 19296129 TI - Brain mythology. PMID- 19296130 TI - Editorial comment: the four principles and cultural specification. AB - The four-principles approach of Beauchamp and Childress has had a huge influence in many areas of medicine, including Pediatrics. There is a risk that such universalist principles fail to take into account cultural differences. This is an important point but there is a need to see the more nuanced aspects of the approach. PMID- 19296131 TI - Analysis of severely affected patients with dihydropyrimidine dehydrogenase deficiency reveals large intragenic rearrangements of DPYD and a de novo interstitial deletion del(1)(p13.3p21.3). AB - Dihydropyrimidine dehydrogenase (DPD) deficiency is an infrequently described autosomal recessive disorder of the pyrimidine degradation pathway and can lead to mental and motor retardation and convulsions. DPD deficiency is also known to cause a potentially lethal toxicity following administration of the antineoplastic agent 5-fluorouracil. In an ongoing study of 72 DPD deficient patients, we analysed the molecular background of 5 patients in more detail in whom initial sequence analysis did not reveal pathogenic mutations. In three patients, a 13.8 kb deletion of exon 12 was found and in one patient a 122 kb deletion of exon 14-16 of DPYD. In the fifth patient, a c.299_302delTCAT mutation in exon 4 was found and also loss of heterozygosity of the entire DPD gene. Further analysis demonstrated a de novo deletion of approximately 14 Mb of chromosome 1p13.3-1p21.3, which includes DPYD. Haploinsufficiency of NTNG1, LPPR4, GPSM2, COL11A1 and VAV3 might have contributed to the severe psychomotor retardation and unusual craniofacial features in this patient. Our study showed for the first time the presence of genomic deletions affecting DPYD in 7% (5/72) of all DPD deficient patients. Therefore, screening of DPD deficient patients for genomic deletions should be considered. PMID- 19296132 TI - Linkage and heritability analysis of migraine symptom groupings: a comparison of three different clustering methods on twin data. AB - Migraine is a painful disorder for which the etiology remains obscure. Diagnosis is largely based on International Headache Society criteria. However, no feature occurs in all patients who meet these criteria, and no single symptom is required for diagnosis. Consequently, this definition may not accurately reflect the phenotypic heterogeneity or genetic basis of the disorder. Such phenotypic uncertainty is typical for complex genetic disorders and has encouraged interest in multivariate statistical methods for classifying disease phenotypes. We applied three popular statistical phenotyping methods-latent class analysis, grade of membership and grade of membership "fuzzy" clustering (Fanny)-to migraine symptom data, and compared heritability and genome-wide linkage results obtained using each approach. Our results demonstrate that different methodologies produce different clustering structures and non-negligible differences in subsequent analyses. We therefore urge caution in the use of any single approach and suggest that multiple phenotyping methods be used. PMID- 19296133 TI - Gene expression profiling of human mesenchymal stem cells chemotactically induced with CXCL12. AB - In situ tissue engineering is a promising approach in regenerative medicine, with the possibility that adult stem or progenitor cells will be guided chemotactically to a tissue defect and subsequently differentiate into the surrounding tissue type. Mesenchymal stem cells (MSC) represent attractive candidate cells. Chemokines such as CXCL12 (SDF-1alpha) chemoattract MSC, but little is known about the molecular processes involved in the chemotaxis and migration of MSC. In this study, MSC recruitment by CXCL12 was investigated by genome-wide microarray analysis. The dose-dependent migration potential of bone marrow-derived MSC toward CXCL12 was measured in an in vitro assay, with a maximum being recorded at a concentration of 1,000 nM CXCL12. Microarray analysis of MSC stimulated with CXCL12 and non-stimulated controls showed 30 differentially expressed genes (24 induced and six repressed). Pathway analysis revealed 11 differentially expressed genes involved in cellular movement and cytokine-cytokine receptor interaction, including those for migratory inducers such as the chemokines CXCL8 and CCL26, the leukocyte inhibitory factor, secretogranin II, and prostaglandin endoperoxide synthase 2. These results were confirmed by real-time polymerase chain reaction for selected genes. The obtained data provide further insights into the molecular mechanisms involved in chemotactic processes in cell migration and designate CXCL12 as a promising candidate for in situ recruitment in regenerative therapies. PMID- 19296134 TI - Overlapping presentation of fungal tubulointerstitial nephritis in an immunosuppressed pediatric patient. AB - With the expanding use of immunosuppressive therapies and broad-spectrum antibiotics, Candida species has become an increasingly important cause of infections, particularly in the presence of anti-tumor necrosis factor-alpha therapy. We report the case of a 17-year-old female with ulcerative colitis who developed oliguric renal failure following immunosuppressive and nephrotoxic therapy. Although urine cultures and urinary tract imaging were negative in the face of fungemia, renal biopsy was the key to establishing the diagnosis of fungal tubulo-interstitial nephritis as the primary reversible cause of the renal failure. PMID- 19296135 TI - Chlorhexidine-based antiseptic solutions effectively reduce catheter-related bacteremia. AB - The aim of this retrospective study was to investigate if the application of chlorhexidine-based solutions (ChloraPrep) to the exit site and the hub of long term hemodialysis catheters could prevent catheter-related bacteremia (CRB) and prolong catheter survival when compared with povidone-iodine solutions. There were 20,784 catheter days observed. Povidone-iodine solutions (Betadine) were used in the first half of the study and ChloraPrep was used in the second half for all the patients. Both groups received chlorhexidine-impregnated dressings at the exit sites. The use of ChloraPrep significantly decreased the incidence of CRB (1.0 vs 2.2/1,000 catheter days, respectively, P = 0.0415), and hospitalization due to CRB (1.8 days vs 4.1 days/1,000 catheter days, respectively, P = 0.0416). The incidence of exit site infection was similar for the two groups. Both the period of overall catheter survival (207.6 days vs 161.1 days, P = 0.0535) and that of infection-free catheter survival (122.0 days vs 106.9 days, P = 0.1100) tended to be longer for the catheters cleansed with ChloraPrep, with no statistical significance. In conclusion, chlorhexidine-based solutions are more effective for the prevention of CRB than povidone-iodine solutions. This positive impact cannot be explained by decreased number of exit site infections. This study supports the notion that the catheter hub is the entry site for CRB. PMID- 19296136 TI - The quality of spine surgery from the patient's perspective: part 2. Minimal clinically important difference for improvement and deterioration as measured with the Core Outcome Measures Index. AB - The Core Outcome Measures Index (COMI) is a reliable and valid instrument for assessing multidimensional outcome in spine surgery. The minimal clinically important score-difference (MCID) for improvement (MCID(imp)) was determined in one of the original research studies validating the instrument, but has never been confirmed in routine clinical practice. Further, the MCID for deterioration (MCID(det)) has never been investigated; indeed, this needs very large sample sizes to obtain sufficient cases with worsening. This study examined the MCIDs of the COMI in routine clinical practice. All patients undergoing surgery in our Spine Center since February 2004 were asked to complete the COMI before and 12 months after surgery. The COMI has one question each on back (neck) pain intensity, leg/buttock (arm/shoulder) pain intensity, function, symptom-specific well-being, general quality of life, work disability, and social disability, scored as a 0-10 index. At follow-up, patients also rated the global effectiveness of surgery, on a 5-point Likert scale. This was used as the external criterion ("anchor") in receiver operating characteristics (ROC) analyses to derive cut-off scores for individual improvement and deterioration. Twelve-month follow-up questionnaires were returned by 3,056 (92%) patients. The group mean COMI score change for patients declaring that the "operation helped" was a reduction of 3.1 points; the corresponding value for those whom it "did not help" was a reduction of 0.5 points. The group MCID(imp) was hence 2.6 points reduction; the corresponding group MCID(det) was 1.2 points increase (0.5 minus 0.7). The area under the ROC curve was 0.88 for MCID(imp) and 0.89 for MCID(det) (both P < 0.0001), indicating that the COMI had good discriminative ability. The cut-offs for individual improvement and deterioration, respectively, were > or =2.2 points decrease (sensitivity 81%, specificity 83%) and > or =0.3 points increase (sensitivity 83%, specificity 88%). The MCID(imp) score of 2.2 points was similar to that reported in the original study (2-3 points, depending on external criterion used). The MCID(det) suggested that the COMI is less responsive to deterioration than to improvement, a phenomenon also reported for other spine outcome instruments. This needs further investigation in even larger patient groups. The MCIDs provide essential information for both the planning (sample size) and interpretation of the results (clinical relevance) of future clinical studies using the COMI. PMID- 19296137 TI - Susceptibility of clinical isolates of frequently encountered bacterial species to tigecycline one year after the introduction of this new class of antibiotics: results of the second multicentre surveillance trial in Germany (G-TEST II, 2007). AB - Tigecycline, a broad-spectrum antibiotic for parenteral use, was introduced in Germany in May 2006. In the G-TEST-II trial, the susceptibility of isolates, recovered in 2007 from hospitalised patients in 15 centres, was assessed against tigecycline and comparators. Susceptibility tests were performed by the microdilution procedure. This study reports on the susceptibility of the isolates of 16 bacterial species and compares the results with those of a trial (G-TEST I) conducted prior to the introduction of tigecycline. Between 2005 and 2007, tigecycline retained activity against Gram-positive and Gram-negative organisms. By contrast, the rate of vancomycin-resistant strains among Enterococcus faecium isolates almost doubled. Moreover, an increase in resistance to broad-spectrum beta-lactams and fluoroquinolones was observed for members of the family Enterobacteriaceae. Against a background of a steadily rising number of pathogens that are resistant to various antibiotic classes, tigecycline represents an important treatment option. PMID- 19296138 TI - Correlation of incidental colorectal wall thickening at CT compared to colonoscopy. AB - Incidental colon wall thickening is a common finding on computed tomography (CT) scans obtained in patients presenting to the emergency department. Currently, there are no guidelines for the emergency department physician or primary care physician regarding the need for further evaluation, specifically colonoscopy, in these patients. A retrospective review of 2,850 abdominopelvic CT scans performed from April 2006 to September 2006 was conducted. Three hundred twenty-two patients had incidental colon wall thickening and 150 of these patients had a subsequent colonoscopy. This study focused on those 150 patients and analyzed the correlation between the CT scan findings of wall thickening of the colon and the findings on colonoscopy. The CT scan findings correlated with abnormal colonoscopic findings in 96 out of 150 (64%) of our cases. In a specific subgroup, 12 out of 12 (100%) of these CT scans were also noted to have a mass lesion, as well as colon wall thickening, all of which were consistent with an adenoma or carcinoma found on colonoscopy. Incidental colorectal wall thickening at CT scan is due to nonspecific colitis in majority of the cases. Admission to the hospital should be based upon criteria other than this incidental radiological finding. However, patients with colorectal wall thickening with a mass lesion at CT scan should be followed closely as inpatient or outpatient given the high correlation with significant findings on colonoscopy. PMID- 19296139 TI - Cost-effectiveness of immunosuppressive regimens in renal transplant recipients in Germany: a model approach. AB - BACKGROUND: The choice of immunosuppression regimen is of paramount importance for outcomes and cost of renal transplantation. We compared the cost effectiveness of triple immunosuppressive regimens in Germany. METHODS: A strong micro-simulation model was built comparing regimens based on cyclosporine, everolimus, sirolimus, and tacrolimus. Mean cost per patient, incremental cost per life year gained, and incremental cost per additional year with functioning graft were assessed from the perspective of the German statutory health insurance (SHI) after 2 and 10 years. RESULTS: Over the 2-year period, the model predicted mean total costs per patient of 26,732, 29,352, 33,415, and 49,978 euro for sirolimus, cyclosporine, everolimus, and tacrolimus, respectively. Focusing on the cost per life year gained, the sirolimus-based regimen compared favorably with those based on everolimus and tacrolimus. The incremental cost-effectiveness ratio (ICER) of cyclosporine versus sirolimus is 524,000 euro per life year gained. Regarding the cost per year with functioning graft gained, sirolimus dominated cyclosporine and everolimus, while the ICER for tacrolimus compared to sirolimus amounts to 1,788,154 euro. Over the 10-year time frame, mean total costs per patient were 100,758, 108,300, 120,316, and 183,802 euro for sirolimus, cyclosporine, everolimus, and tacrolimus, respectively. With regard to life years gained, sirolimus dominated both cyclosporine and everolimus. The ICER of tacrolimus versus sirolimus was 1,766,894 euro. Considering the years with functioning graft gained, sirolimus dominated cyclosporine and everolimus, while the ICER for tacrolimus compared to sirolimus amounted to 1,339,419 euro. CONCLUSIONS: Over both the 2-year and the 10-year time horizon, sirolimus-based immunosuppression represents a cost-effective option in renal transplantation in Germany. PMID- 19296140 TI - AhR agonist and genotoxicant bioavailability in a PAH-contaminated soil undergoing biological treatment. AB - BACKGROUND, AIM, AND SCOPE: Degradation of the 16 US EPA priority PAHs in soil subjected to bioremediation is often achieved. However, the PAH loss is not always followed by a reduction in soil toxicity. For instance, bioanalytical testing of such soil using the chemical-activated luciferase gene expression (CALUX) assay, measuring the combined effect of all Ah receptor (AhR) activating compounds, occasionally indicates that the loss of PAHs does not correlate with the loss of Ah receptor-active compounds in the soil. In addition, standard PAH analysis does not address the issue of total toxicant bioavailability in bioremediated soil. MATERIALS AND METHODS: To address these questions, we have used the CALUX AhR agonist bioassay and the Comet genotoxicity bioassay with RTL W1 cells to evaluate the toxic potential of different extracts from a PAH contaminated soil undergoing large-scale bioremediation. The extracts were also chemically analyzed for PAH16 and PCDD/PCDF. Soil sampled on five occasions between day 0 and day 274 of biological treatment was shaken with n-butanol with vortex mixing at room temperature to determine the bioavailable fraction of contaminants. To establish total concentrations, parts of the same samples were extracted using an accelerated solvent extractor (ASE) with toluene at 100 degrees C. The extracts were tested as inducers of AhR-dependent luciferase activity in the CALUX assay and for DNA breakage potential in the Comet bioassay. RESULTS: The chemical analysis of the toluene extracts indicated slow degradation rates and the CALUX assay indicated high levels of AhR agonists in the same extracts. Compared to day 0, the bioavailable fractions showed no decrease in AhR agonist activity during the treatment but rather an up-going trend, which was supported by increasing levels of PAHs and an increased effect in the Comet bioassay after 274 days. The bio-TEQs calculated using the CALUX assay were higher than the TEQs calculated from chemical analysis in both extracts, indicating that there are additional toxic PAHs in both extracts that are not included in the chemically derived TEQ. DISCUSSION: The response in the CALUX and the Comet bioassays as well as the chemical analysis indicate that the soil might be more toxic to organisms living in soil after 274 days of treatment than in the untreated soil, due to the release of previously sorbed PAHs and possibly also metabolic formation of novel toxicants. CONCLUSIONS: Our results put focus on the issue of slow degradation rates and bioavailability of PAHs during large-scale bioremediation treatments. The release of sorbed PAHs at the investigated PAH contaminated site seemed to be faster than the degradation rate, which demonstrates the importance of considering the bioavailable fraction of contaminants during a bioremediation process. RECOMMENDATIONS AND PERSPECTIVES: It has to be ensured that soft remediation methods like biodegradation or the natural remediation approach do not result in the mobilization of toxic compounds including more mobile degradation products. For PAH-contaminated sites this cannot be assured merely by monitoring the 16 target PAHs. The combined use of a battery of biotests for different types of PAH effects such as the CALUX and the Comet assay together with bioavailability extraction methods may be a useful screening tool of bioremediation processes of PAH-contaminated soil and contribute to a more accurate risk assessment. If the bioremediation causes a release of bound PAHs that are left undegraded in an easily extracted fraction, the soil may be more toxic to organisms living in the soil as a result of the treatment. A prolonged treatment time may be one way to reduce the risk of remaining mobile PAHs. In critical cases, the remediation concept might have to be changed to ex situ remediation methods. PMID- 19296141 TI - Levels and homologue profiles of PCDD/Fs in sediments along the Swedish coast of the Baltic Sea. AB - BACKGROUND, AIM, AND SCOPE: The primary aim of this study was to explore the variations in PCDD/F levels and homologue profiles of Baltic surface sediments by comprehensively analyzing polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) in samples from a large number of sites, encompassing not only previously known hotspot areas, but also sites near other potential PCDD/F sources, in pristine reference areas (in which there was no industrial activity) and offshore sites. MATERIALS AND METHODS: Surface sediment samples (146 in total) were collected at various points along the Swedish coast and offshore areas. In addition, bulk deposition was sampled, monthly, at a single site in northern Sweden during 1 year. The concentrations of tetra- through octa substituted CDD/Fs were determined in both matrices. RESULTS: Highly elevated concentrations of PCDD/Fs were found at many sites in coastal areas and concentrations were also slightly elevated in some offshore areas. Homologue profiles varied substantially amongst samples from coastal sites, while those from offshore and other pristine sediments were relatively similar. The offshore sediments showed different profiles from those observed in the deposition samples. Sediment levels of PCDD/Fs were not generally significantly correlated to organic carbon levels, except in some pristine areas. Comparison of data obtained in this and previous studies suggest that both their levels and profiles are similar today to those observed 20 years ago in coastal and offshore areas. The only detected trend is that their levels appear to have decreased slightly in the offshore area of the Bothnian Sea. DISCUSSION: The localization of hotspot areas along the coast, the lack of consensus between PCDD/F profiles of sediments and general background, and their weak correlations with organic carbon suggest that PCDD/Fs in the study area largely originate from local/regional emissions. However, due to complicating factors such as sediment dynamics and land upheaval, it is not possible to conclude whether these pollutants derive from recent emissions or from a combination of recent emissions and re-distribution of previous inputs. CONCLUSIONS: The results show that: elevated levels of PCDD/Fs are present in both coastal and offshore areas of the Baltic Sea, the major hotspots are close to the shore, and there are large variations in profiles, indicating that local emissions are (or have been) the major causes of pollution. RECOMMENDATIONS AND PERSPECTIVES: In order to identify other hotspot areas and trace sources, comprehensive analysis of PCDD/Fs in surface sediments is needed in all areas of the Baltic Sea that have not been previously investigated. The high levels of PCDD/Fs observed in surface sediments also indicate a need to elucidate whether they are due mainly to current emissions or a combination of recent pollution and re-distribution of historically deposited pollutants. To do so, better understanding of sediment dynamics and present-day inputs, such as riverine inputs, industrial effluents, and leakage from contaminated soil is required. There are indications that contaminated sediments have a regional impact on fish contamination levels. However, as yet there is no statistically robust evidence linking contaminated sediments with elevated levels in Baltic biota. It should also be noted that the Baltic Sea is being massively invaded by the deep-burrowing polychaete Marenzielleria ssp., whose presence in sediments has been shown to increase water concentrations of hydrophobic pollutants. In awareness of this, it is clear that high levels in sediments cannot be ignored in risk assessments. In order to investigate the emission trends more thoroughly, analysis of PCDD/Fs in offshore sediment cores throughout the Baltic Sea is also recommended. PMID- 19296142 TI - A validated finite element analysis of nerve root stress in degenerative lumbar scoliosis. AB - Few studies have shown the relationship between the curve pattern and nerve root symptoms in degenerative lumbar scoliosis, and its mechanism remains unclear. We developed a finite element model of two patterns of scoliotic curves (isolated lateral bending curve, lateral bending combined with rotation curve). The stress on the nerve root was calculated on both sides (right and left) of the apex vertebra. In the lateral bending curves without rotation, the compressive nerve root stress on the concave side was greater than the tensile stress on the convex side at the apex vertebra. In contrast, when the segmental rotation of the vertebrae was added to the lateral bending curve, there was significantly higher tensile stress on the convex side, and lower compressive stress on the concave side. To conclude, rotatory listhesis may be an important pathomechanism in the development of neurologic symptoms on the convex side of the curve. PMID- 19296143 TI - European Society of Neuroradiology (ESNR) : Board, committees, and meeting information. PMID- 19296144 TI - Risedronate dosing before breakfast compared with dosing later in the day in women with postmenopausal osteoporosis. AB - Two studies in postmenopausal women with osteoporosis provide information about the efficacy and safety of dosing oral risedronate 5 mg daily at a time other than before breakfast (i.e., 2 h before and 2 h after any food and drink other than plain water). A significant increase in lumbar spine BMD was observed for both treatment regimens in the two studies. However, smaller increases in lumbar spine BMD were observed with flexible dosing versus before-breakfast dosing. Geographic region, compliance, and consistency of dosing time appear to affect the amount of increase in BMD observed with flexible dosing. INTRODUCTION: Two studies in postmenopausal women with osteoporosis provide additional information about the efficacy and safety of dosing oral risedronate 5 mg daily at a time other than before breakfast (i.e., 2 h before and 2 h after any food and drink other than plain water). METHODS: One study, flexible dosing, was a 6-month North American study in 730 patients randomized to before-breakfast dosing or flexible dosing later in the day. A second study, IMPACT, was a large (N = 2382), 1-year multinational study in patients that chose their dosing regimen (before breakfast or later in the day). These studies were used to examine the bone mineral density (BMD) response with different dosing regimens. RESULTS: A significant increase in lumbar spine BMD was observed for both treatment regimens in the two studies. However, in both studies, the flexible dosing group had a smaller increase from baseline compared to the before-breakfast regimen (ratio of flexible dosing to before breakfast: flexible dosing study, 0.52; IMPACT study, 0.75). In addition, a relationship between geographic region and BMD response was observed with flexible dosing in both studies. Patients in the flexible dosing group who had greater dosing compliance (based on the number of times the bottle was opened) and consistency of dosing time (bottle opened within a 1.5-h window) had a greater increase in lumbar spine BMD. CONCLUSION: Results of these two studies demonstrate that overall flexible dosing of risedronate leads to smaller BMD gains compared to before-breakfast dosing. This result may be due to poorer adherence to the flexible dosing instructions that may be more pronounced in patients in certain geographic regions. If patients cannot abide by before breakfast dosing and flexible dosing is an approved option, one can expect suboptimal BMD results with flexible dosing. PMID- 19296145 TI - Analysis of routine EEG usage in a general adult ICU. AB - BACKGROUND: Non-convulsive seizures and status epilepticus are common in brain injured patients in intensive care units. Continuous electroencephalography (cEEG) monitoring is the most sensitive means of their detection. In centres where cEEG is unavailable, routine EEG is often utilized for diagnosis although its sensitivity is lower. AIMS: To establish the rate of electrographic seizure detection in ICU using routine EEG. METHODS: We identified all routine EEGs performed within a general adult ICU in Ireland over 3 years, and analyzed the clinical and EEG data. RESULTS: Fifty-two patients underwent single or repeated EEG evaluation during the time period. Epileptiform abnormalities were evident in 15%, periodic abnormalities in 14%, and electrographic seizures in just one patient (2%) in their first or only routine EEG recording. CONCLUSION: The rate of electrographic seizure detection by routine EEG in a general ICU is lower than anticipated. Earlier and more prolonged recordings are needed in this setting. PMID- 19296146 TI - Myopericarditis following Campylobacter infection. AB - BACKGROUND: Myopericarditis can be caused by a wide range of infectious agents. There are a few reported cases implicating Campylobacter in the pathogenesis of myopericarditis. CONCLUSIONS: We believe that this case report represents the first documented case in Ireland of myopericarditis associated with Campylobacter enteritis. PMID- 19296147 TI - Surgical and laser treatment of Sturge-Weber syndrome. AB - Sturge-Weber syndrome (also called encephalofacial or encephalotrigeminal angiomatosis) is a rare neurocutaneous syndrome characterized by facial port-wine stains in the trigeminal nerve distribution area. An innovative treatment based on surgical and laser techniques of a patient affected with Sturge-Weber syndrome is reported. The aesthetic and functional results were satisfactory. PMID- 19296148 TI - A case of late unilateral hematoma and subsequent late seroma of the breast after bilateral breast augmentation. AB - BACKGROUND: A previously unreported case of unilateral recurrent late collections, namely, a hematoma and a subsequent seroma, in an otherwise healthy female patient after breast augmentation is presented. METHODS: A case is reported together with a review of the world literature published and presented regarding late postoperative breast augmentation complications. RESULTS: The patient was a 49-year-old woman who underwent bilateral breast augmentation in 2006. Spontaneous swelling of the left breast developed 1 year postoperatively. The patient was taken to the operating theater, where a large hematoma was found and drained and the prosthesis reinserted. No cause for the complication was identified at the operation. The patient's symptoms of left-sided breast swelling with discomfort recurred 9 months later. A large seroma was found at surgery but no blood staining. On both occasions, the implant was extracted and examined, with no fault discovered. Capsulectomy was performed, and the pocket was examined, debrided, and cauterized as necessary. On neither occasion was any remarkable pathology found clinically or in laboratory samples sent for analysis. The patient was investigated for any sensitivities or hematologic abnormalities, with no significant results. CONCLUSION: A previously unreported case of recurrent late postaugmentation collections, namely, a seroma and a subsequent hematoma, in an otherwise well female patient is reported. The authors found no obvious reason for the recurrence in this patient. PMID- 19296149 TI - Breast augmentation in asymmetrically placed nipple-areola complex in the horizontal axis: lateralisation of implant pocket to offset lateralised nipples. AB - BACKGROUND: Nipple-areola complex (NAC) lateralisation in the horizontal axis is understudied with respect to its potential effects on the outcome of augmentation mammoplasty. An implant pocket dissected slightly laterally on the affected side offsets the lateralised nipple and improves the quality of breast enhancement in the lateral quadrant with an aesthetically positioned NAC. It also prevents the appearance of medially malplaced implants. METHODS: Medial and lateral halves of breasts were measured in 312 mammaplasties performed by the author in 2007; of these, 39 (12%) had NAC asymmetrically positioned in the horizontal plane, 29 (9%) were right lateralised, 4 (1.2%) left lateralised, 5 (1.5%) bilaterally lateralised, and 1 (0.3%) was bilaterally medialised. In the more common right lateralised group (n = 25), the mean medial half measurement was 8.76 +/- 1.22 cm compared with 8.96 cm in the control group (n = 170), with no statistical significance (P = 0.45). However, the lateral half measurement in the lateralised nipple group (n = 25) was 9.06 +/- 1.53 cm compared with 9.7 +/- 1.67 cm in the control group (n = 169) and it showed a borderline significant value (P = 0.06). On the affected side, the medial boundary of the breast pocket is marked 0.25-0.5 cm further away from the routine 2.5-cm intermammary distance with the goal of offsetting the lateralised NAC and to improve its visual aesthetic position on the breast. RESULTS AND CONCLUSION: Lateralisation of the breast pocket in a breast with a lateralised nipple improves the aesthetic position of the NAC. In compensated augmentation mammaplasties, lateral quadrant enhancement of the breast is aesthetically superior to uncompensated cases and prevents the appearance of medially malplaced implants. The intermammary distance in compensated cases was acceptable without any compromise to the aesthetic outcome. PMID- 19296150 TI - Correction of recurrent grade III inverted nipple with antenna dermoadipose flap: case report. AB - BACKGROUND: Numerous techniques have been reported for the correction of the inverted nipple. It was also reported that no single technique was appropriate for correcting all types of nipple deformities because different grades of inverted nipple have different levels of fibrosis, soft tissue bulk, and lactiferous ductus structure. In this article a novel technique for the repair of recurrent grade III inverted nipple is described in detail. METHODS: In this technique, dermoadipose flaps were generated within the area of de epithelialization of mastopexy.The flap is called "antenna flap" because of its designing. RESULTS: No major vital complications such as necrosis of the major flap or nipple were seen. There was no recurrence of nipple inversion after 15 months. The shape and projection of the patient's nipple was deemed satisfactory. CONCLUSION: This technique yielded a satisfactory result without recurrence of inversion. It should be considered for use on any patient contemplating correction of inverted nipple and mastopexy. It should be emphasized that this technique also permits the performance of mastopexy in conjunction with the correction of inverted nipple. PMID- 19296151 TI - Reoperation for the removal of polyacrylamide hydrogel in the breast: use of periareolar approach under direct visualization. PMID- 19296152 TI - Lipoabdominoplasty: revisiting the superior pull-down abdominal flap and new approaches. AB - Abdominoplasty is a very common procedure, especially for patients with abdominal laxness, striaes, and muscle rectus diastases. With the advent of liposuction 28 years ago, we can improve body contouring by treating lipodystrophies in the epigastric, flank, trochanteric, and buttocks areas. The procedure combining abdominoplasty and liposuction is called lipoabdominoplasty. Many new techniques have been proposed since these procedures were introduced; we now revisit the superior pull-down abdominal flap technique with several new modifications and improvements. PMID- 19296153 TI - Clinical efficacy of noninvasive cryolipolysis and its effects on peripheral nerves. AB - BACKGROUND: Cryolipolysis provides a method for noninvasive fat reduction that significantly reduces subcutaneous fat in a pig model without apparent damage to skin and surrounding structures. This study aimed to determine whether fat reduction in humans caused by cold exposure is associated with alteration in local sensory function or nerve fibers. METHODS: In this study, 10 subjects were treated with a prototype cooling device. Fat reduction was assessed in 9 of the 10 subjects via ultrasound before treatment and at the follow-up visit. Sensory function was assessed by neurologic evaluation (n = 9), and biopsies (n = 1) were collected for nerve staining. RESULTS: Treatment resulted in a normalized fat layer reduction of 20.4% at 2 months and 25.5% at 6 months after treatment. Transient reduction in sensation occurred in six of nine subjects assessed by neurologic evaluation. However, all sensation returned by a mean of 3.6 weeks after treatment. Biopsies showed no long-term change in nerve fiber structure. There were no lasting sensory alterations or observations of skin damage in any of the subjects evaluated. CONCLUSION: Noninvasive cryolipolysis results in substantial fat reduction within 2 months of treatment without damage to skin. The procedure is associated with modest reversible short-term changes in the function of peripheral sensory nerves. PMID- 19296154 TI - Quantifying terrestrial ecosystem carbon dynamics in the Jinsha watershed, upper Yangtze, China from 1975 to 2000. AB - Quantifying the spatial and temporal dynamics of carbon stocks in terrestrial ecosystems and carbon fluxes between the terrestrial biosphere and the atmosphere is critical to our understanding of regional patterns of carbon budgets. Here we use the General Ensemble biogeochemical Modeling System to simulate the terrestrial ecosystem carbon dynamics in the Jinsha watershed of China's upper Yangtze basin from 1975 to 2000, based on unique combinations of spatial and temporal dynamics of major driving forces, such as climate, soil properties, nitrogen deposition, and land use and land cover changes. Our analysis demonstrates that the Jinsha watershed ecosystems acted as a carbon sink during the period of 1975-2000, with an average rate of 0.36 Mg/ha/yr, primarily resulting from regional climate variation and local land use and land cover change. Vegetation biomass accumulation accounted for 90.6% of the sink, while soil organic carbon loss before 1992 led to a lower net gain of carbon in the watershed, and after that soils became a small sink. Ecosystem carbon sink/source patterns showed a high degree of spatial heterogeneity. Carbon sinks were associated with forest areas without disturbances, whereas carbon sources were primarily caused by stand-replacing disturbances. It is critical to adequately represent the detailed fast-changing dynamics of land use activities in regional biogeochemical models to determine the spatial and temporal evolution of regional carbon sink/source patterns. PMID- 19296155 TI - Whole-body magnetic resonance angiography at 3 tesla using a hybrid protocol in patients with peripheral arterial disease. AB - The purpose of this study was to determine the diagnostic performance of 3T whole body magnetic resonance angiography (WB-MRA) using a hybrid protocol in comparison with a standard protocol in patients with peripheral arterial disease (PAD). In 26 consecutive patients with PAD two different protocols were used for WB-MRA: a standard sequential protocol (n = 13) and a hybrid protocol (n = 13). WB-MRA was performed using a gradient echo sequence, body coil for signal reception, and gadoterate meglumine as contrast agent (0.3 mmol/kg body weight). Two blinded observers evaluated all WB-MRA examinations with regard to presence of stenoses, as well as diagnostic quality and degree of venous contamination in each of the four stations used in WB-MRA. Digital subtraction angiography served as the method of reference. Sensitivity for detecting significant arterial disease (luminal narrowing > or = 50%) using standard-protocol WB-MRA for the two observers was 0.63 (95%CI: 0.51-0.73) and 0.66 (0.58-0.78). Specificities were 0.94 (0.91-0.97) and 0.96 (0.92-0.98), respectively. In the hybrid protocol WB MRA sensitivities were 0.75 (0.64-0.84) and 0.70 (0.58-0.8), respectively. Specificities were 0.93 (0.88-0.96) and 0.95 (0.91-0.97). Interobserver agreement was good using both the standard and the hybrid protocol, with kappa = 0.62 (0.44 0.67) and kappa = 0.70 (0.59-0.79), respectively. WB-MRA quality scores were significantly higher in the lower leg using the hybrid protocol compared to standard protocol (p = 0.003 and p = 0.03, observers 1 and 2). Distal venous contamination scores were significantly lower with the hybrid protocol (p = 0.02 and p = 0.01, observers 1 and 2). In conclusion, hybrid-protocol WB-MRA shows a better diagnostic performance than standard protocol WB-MRA at 3 T in patients with PAD. PMID- 19296156 TI - Image-guided radiofrequency ablation of a pancreatic tumor with a new triple spiral-shaped electrode. AB - Image-guided, minimally invasive treatment modalities have become an area of considerable interest and research during the last few years for the treatment of primary and secondary liver tumors. We report our experience with an unresectable pancreatic tumor, treated with application of radiofrequency ablation under CT guidance that even though a complication occurred during the procedure, had excellent results on follow-up CT scans. PMID- 19296157 TI - Hepatic arterial infusion chemotherapy using fluorouracil followed by systemic therapy using oxaliplatin plus fluorouracil and leucovorin for patients with unresectable liver metastases from colorectal cancer. AB - The purpose of this study was to assess retrospectively the sequential treatment of hepatic arterial infusion (HAI) chemotherapy followed by systemic therapy using oxaliplatin plus 5-fluorouracil (5-FU) and leucovorin, namely, FOLFOX, for patients with liver metastases from colorectal cancer. We reviewed 20 patients with unresectable liver metastases from colorectal cancer. Patients were initially treated with HAI chemotherapy until disease progression (5 fluorouracil, 1000 mg/m(2) intra-arterial infusion, weekly) and then with FOLFOX thereafter (FOLFOX4, n = 13; modified FOLFOX6, n = 7). Adverse events, tumor response, and time to progression for each therapy were evaluated retrospectively, and overall survival was estimated. Toxicity of HAI chemotherapy was generally mild. Of 20 patients, adverse events leading to treatment discontinuation occurred in only 1 patient (5%) during initial therapy using HAI chemotherapy, while 9 patients (45%) exhibited adverse events during subsequent FOLFOX therapy. For HAI chemotherapy and FOLFOX, objective response rates were 85.0% and 35.0%, respectively, and median time to progression was 11.6 and 5.1 months, respectively. Median overall survival was 30.1 months. In conclusion, the sequence of HAI chemotherapy followed by FOLFOX is a promising treatment strategy for the long-term use of active chemotherapeutic agents, leading to a superior tumor response and fewer toxic effects in patients with unresectable liver metastases from colorectal cancer. PMID- 19296158 TI - Use of intra-arterial chemotherapy and embolization before limb salvage surgery for osteosarcoma of the lower extremity. AB - We report our experience with the use of intra-arterial chemotherapy and embolization before limb salvage surgery in patients with osteosarcoma of the lower extremity. We evaluated the effect of this procedure on the degree of tumor necrosis and on the amount of blood loss during surgery. We reviewed the medical records of all patients who received intra-arterial chemotherapy and embolization before undergoing limb salvage surgery for osteosarcoma of the lower extremity at our institution between January 2003 and April 2008. Patient demographic, tumor characteristics, treatment details, postembolization complications, and surgical and pathological findings were recorded for each patient. We evaluated the operative time, estimated blood loss (EBL), and volume of blood transfusion during surgery and in the postoperative period in all patients in the study group. The same parameters were recorded for 65 other patients with lower extremity osteosarcoma who underwent limb salvage operation at our institution without undergoing preoperative intervention. The study included 47 patients (25 males and 22 females). Angiography showed that the tumors were hypervascular. Intra-arterial chemotherapy and embolization were performed successfully, resulting in a substantial reduction or complete disappearance of tumor stain in all patients. No major complications were encountered. At the time of surgery, performed 3-7 days after embolization, a fibrous edematous band around the tumor was observed in 43 of the 47 patients, facilitating surgery. The goal of limb salvage was achieved successfully in all cases. Percentage tumor necrosis induced by treatment ranged from 70.2% to 94.2% (average, 82.9%). EBL during surgery, EBL from drains in the postoperative period, total EBL, and transfusion volumes were significantly lower in the 47 study patients compared to the 65 patients who underwent surgery without preoperative treatment with intra-arterial chemotherapy and embolization. The mean operative time was also significantly less in the intervention group compared to the nonintervention group (73.2 vs. 88.5 min; p < 0.05). In conclusion, intra-arterial chemotherapy and embolization performed 3 to 7 days before limb salvage surgery in patients with lower extremity osteosarcomas can cause substantial tumor necrosis, reduce the EBL and transfusion requirements during surgery, and induce formation of a false capsule around the tumor, thus facilitating surgical excision of the tumors. PMID- 19296159 TI - Percutaneous management of high-output chylothorax: case reviews. AB - Chylothorax carries significant mortality and morbidity. Patients with high output chylothorax have traditionally been managed by surgical treatment if nonoperative management has proved unsuccessful. Newer, more recent percutaneous techniques used to treat chylothorax are safer and less invasive than surgery. We present three cases that have been successfully managed using these percutaneous techniques. PMID- 19296160 TI - Endovascular repair of a traumatic axillary artery pseudoaneurysm. PMID- 19296161 TI - Improving results of elective abdominal aortic aneurysm repair at a low-volume hospital by risk-adjusted selection of treatment in the endovascular era. AB - Several studies have observed both higher mortality rates and lower utilization of endovascular aneurysm repair (EVAR) at low-volume centers. This article presents the results of elective abdominal aortic aneurysm (AAA) repair at a low volume center in the endovascular era and investigates whether postprocedural mortality can be improved by extension of EVAR application also in this setting. This is an 11.6-year retrospective cohort study of 132 patients undergoing elective surgical or endovascular AAA repair at a tertiary care academic hospital between 1997 and July 2008, i.e., a median volume of 12 cases per year. The study was divided into two periods of time according to the respective indications and contraindications for EVAR, which substantially changed in 2005. During period 1, only aneurysms with necks > or =20 mm long and not involving the iliac arteries were treated endoluminally. Beginning in 2005, indication for EVAR was expanded to aortoiliac aneurysms with a minimum neck length of 15 mm. Preoperative risk was assessed by the SVS/AAVS comorbidity score. During the first period (1997 2004) 18.4% (16/87) of all patients received EVAR. By extending anatomical confines and indications for EVAR in 2005, the utilization rate of EVAR increased to 40.0% (18/45) during the second period (2005-July 2008; p = 0.007). Prevalence of preoperative risk factors did not change during the two observation periods. In contrast to period 1, high-risk patients were preferentially treated endoluminally during the second period, resulting in a significantly higher median SVS/AAVS score in the EVAR group (p < 0.001). A significant decrease in median length of stay at the intensive/intermediate care unit (5 vs. 2 days; p = 0.006) and length of in-hospital stay (20 vs. 12.5 days; p < 0.001) was observed during period 2. Overall perioperative mortality was reduced from 6.9% during the first period to 2.2% during the second period (p = 0.256). EVAR mortality was 0%, mortality after open repair was reduced from 8.5% to 3.7% (p = 0.414). In conclusion, by risk-adjusted selection of treatment and frequent application of EVAR, it is possible to improve perioperative outcome of elective AAA repair at a low-volume hospital. Mortality figures are similar to those of recent trials at high-volume centers, as reported in the literature. PMID- 19296162 TI - Total percutaneous aortic repair: midterm outcomes. AB - The purpose of this study was to examine the immediate and midterm outcomes of percutaneous endovascular repair of thoracic and abdominal aortic pathology. Between December 2003 and June 2005, 21 patients (mean age: 60.4 +/- 17.1 years; 15 males, 6 females) underwent endovascular stent-graft insertion for thoracic (n = 13) or abdominal aortic (n = 8) pathology. Preprocedural computed tomographic angiography (CTA) was performed to assess the suitability of aorto-iliac and common femoral artery (CFA) anatomy, including the degree of CFA calcification, for total percutaneous aortic stent-graft repair. Percutaneous access was used for the introduction of 18- to 26-Fr delivery devices. A 'preclose' closure technique using two Perclose suture devices (Perclose A-T; Abbott Vascular) was used in all cases. Data were prospectively collected. Each CFA puncture site was assessed via clinical examination and CTA at 1, 6, and 12 months, followed by annual review thereafter. Minimum follow-up was 36 months. Outcome measures evaluated were rates of technical success, conversion to open surgical repair, complications, and late incidence of arterial stenosis at the site of Perclose suture deployment. A total of 58 Perclose devices were used to close 29 femoral arteriotomies. Outer diameters of stent-graft delivery devices used were 18 Fr (n = 5), 20 Fr (n = 3), 22 Fr (n = 4), 24 Fr (n = 15), and 26 Fr (n = 2). Percutaneous closure was successful in 96.6% (28/29) of arteriotomies. Conversion to surgical repair was required at one access site (3.4%). Mean follow-up was 50 +/- 8 months. No late complications were observed. By CT criteria, no patient developed a >50% reduction in CFA caliber at the site of Perclose deployment during the study period. In conclusion, percutaneous aortic stent-graft insertion can be safely performed, with a low risk of both immediate and midterm access related complications. PMID- 19296163 TI - A rare case of left ventricular intramural hemangioma diagnosed using 1.5-T cardiac MRI with histopathological correlation and successfully treated by surgery. AB - Hemangiomas are vascular tumors composed of blood vessels, frequently localized in the skin and subcutaneous muscles; their localization in the heart is exceptional. The most common localizations are the lateral walls of the left ventricle, the anterior wall, and the septum. Mostly, these tumors grow intracavitarily, rarely intramurally. We describe a singular case of left ventricular intramural hemangioma, detected and diagnosed using newer magnetic resonance imaging (MRI) modalities, confirmed by histopathological results, and treated successfully by surgery. PMID- 19296164 TI - Characterization of temperate Lactobacillus gasseri phage LgaI and its impact as prophage on autolysis of its lysogenic host strains. AB - We show by electron microscopy that Lactobacillus gasseri phage LgaI, a temperate phage residing in the chromosome of Lactobacillus gasseri ATCC33323, belongs to the family of Myoviridae phages. The LgaI DNA is packed by the "head-full" mechanism, as demonstrated by analysis of restriction patterns of heated (74 degrees C) or non-heated DNA. By isolating prophage-cured cells, we were able to demonstrate phage LgaI to be responsible for the strong autolytic phenotype observed for Lactobacillus gasseri ATCC33323. In addition, we show that a copy of the LgaI prophage resides in the chromosome of Lactobacillus gasseri NCK102. The LgaI prophage was not inducible in L. gasseri NCK102-adh by mitomycin C, however, it apparently contributed to the autolytic phenotype of this strain. PMID- 19296165 TI - Endoscopic treatment of Roux-en-Y gastric bypass-related gastrocutaneous fistulas using a novel biomaterial. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGB) is amongst the commonest surgical intervention for weight loss in obese patients. Gastrocutaneous fistula, which usually occurs along the vertical staple line of the pouch, is amongst its most alarming complications. Medical management comprised of wound drainage, nutritional support, acid suppression, and antibiotics may be ineffective in as many as a third of patients with this complication. We present outcomes after endoscopic application of SurgiSIS, which is a novel biomaterial for the treatment of this complication. DESIGN: A case series of 25 patients. METHODS: Twenty-five patients who had failed conservative medical management of gastrocutaneous fistula after RYGB underwent endoscopic application of SurgiSIS- an acellular fibrogenic matrix biomaterial to help fistula healing. MAIN OUTCOME MEASURES: Fistula closure as assessed by upper gastrointestinal imaging and endoscopic examination. RESULTS: In patients who had failed medical management lasting 4-25 (median, 7) weeks, closure of the fistulous tract was successful after one application in six patients (30%), two applications in 11 patients (55%), and three applications in three patients (15%). There were no procedure related complications. CONCLUSIONS: Endoscopic application of SurgiSIS-an acellular fibrogenic matrix--is safe and effective for the treatment of gastrocutaneous fistula after RYGB. PMID- 19296166 TI - Which term is better: SILS, SPA, LESS, E-NOTES, or TUES? PMID- 19296167 TI - Outcomes of thoracoscopic management of secondary pneumothorax in patients with COPD and interstitial pulmonary fibrosis. AB - BACKGROUND: We retrospectively analyzed the outcomes of surgical treatment for patients with secondary spontaneous pneumothorax. METHODS: Among consecutive patients with secondary pneumothorax who had undergone surgical treatment from 1993 to 2007, those with chronic obstructive pulmonary diseases (COPD) and those with diffuse pulmonary fibrotic diseases (PFD) were collected and divided into two groups (COPD group and PFD group). Postoperative morbidity and mortality were analyzed between the two groups. RESULTS: We enrolled 72 patients (73 surgeries) as the COPD group and 14 patients (14 surgeries) as the PFD group. All of the surgical treatments were initiated through thoracoscopy. Mean age of the patients at surgery was significantly older in the COPD group compared with the PFD group. The surgeries in the COPD group were significantly longer than those in the PFD group. The bleeding volume during surgery in the COPD group was higher than that in the PFD group. Thoracoscopy was more frequently replaced with open thoracotomy in the PFD group (21.4%) than in the COPD group (2.7%; p = 0.0019). In the PFD group, two patients died from postoperative exacerbation of the pulmonary fibrosis and one died from the sepsis caused by the empyema. One patient in the COPD group died of pneumonia. The postoperative mortality rate was significantly higher in the PFD group (21.4%) than in the COPD group (1.4%; p = 0.001). CONCLUSIONS: Favorable results were obtained in patients in the COPD group. However, the postoperative mortality rate in the PFD group was significantly higher than those in the COPD group. This increase in the mortality rate in the PFD group was mainly caused by postoperative exacerbation of pulmonary fibrotic diseases. PMID- 19296168 TI - Primary access-related complications in laparoscopic cholecystectomy via the closed technique: experience of a single surgical team over more than 15 years. AB - BACKGROUND: Laparoscopic cholecystectomy (LC), a common laparoscopic procedure, is a relatively safe invasive procedure, but complications can occur at every step, starting from creation of the pneumoperitoneum. Several studies have investigated procedure-related complications, but the primary access- or trocar related complications generally are underreported, and their true incidence may be higher than studies show. Major vascular or visceral injury resulting from blind access to the abdominal cavity, although rare, has been reported. Of the two methods for creating pneumoperitoneum, the open access technique is reported to have the lower incidence of these injuries. The authors report their experience with the closed method and show that if performed with proper technique, it can be as rapid and safe as other techniques. However, injuries still happen, and the search for the predisposing factors must be continued. METHODS: Between January 1992 and December 2007, a retrospective study examined 15,260 cases of LC performed for symptomatic gallstone disease in the authors' institution by a single team of surgeons. The primary access-related injuries in these cases were retrospectively analyzed. RESULTS: In 15,260 cases of LC, 63 cases of primary access-related complications were identified, for an overall incidence of 0.41%. Major injuries in 11 cases included major vascular and visceral injuries, and minor injuries in 52 cases included omental and subcutaneous emphysema. For the closed method, the findings showed an overall incidence of 0.14% for primary access-related vascular injuries and 0.07% for visceral injuries. CONCLUSION: Primary access-related complications during LC are common and can prove to be fatal if not identified early. The incidence of these injuries with closed methods is no greater than with open methods. No evidence suggests abandonment of the closed-entry method in laparoscopy. PMID- 19296169 TI - An apical symphysial technique using a wide absorbable mesh placed on the apex for primary spontaneous pneumothorax. AB - BACKGROUND: The outcome of thoracoscopic bullectomy for primary spontaneous pneumothorax (PSP) is not satisfactory. To reduce postoperative pneumothorax recurrence after thoracoscopic bullectomy, an effective and easy surgical method is required. We tried a new method using an absorbable mesh that covered the whole apical visceral pleura. METHODS: A total of 157 sides of 143 patients who underwent stapled bullectomy under thoracoscopy for PSP were reviewed retrospectively. In the apical covering group (group A), a 15 x 15-cm(2) absorbable mesh sheet was placed on the apical visceral pleura with fibrin glue. Patients in group B underwent bullectomy alone. Cumulative postoperative recurrence was compared between the groups. Recurrent cases in group A were examined clinicopathologically. RESULTS: Group A had 111 cases and group B had 46. There was no operative mortality. Postoperative recurrence occurred in 15 of 157 cases (9.6%): 4 in group A and 11 in group B. The cumulative postoperative 5 year recurrence rate was 3.6% in group A and 23.9% in group B (log-rank test, p = 0.013). In group A, local adhesion was seen at the apical pleurae, and inflammatory changes with foreign body giant cells were seen at the pleura covered with the mesh. CONCLUSIONS: Placement of a wide absorbable mesh with fibrin glue at the apical visceral pleura significantly reduced postoperative recurrence after thoracoscopic bullectomy for PSP. The mesh was thought to act as a foreign body on the pleura and induce local inflammatory adhesion between the apical pleurae after bullectomy. This was an easy and effective symphysial procedure. PMID- 19296170 TI - Time-dependent morphologic change in depressed-type early gastric cancer. AB - BACKGROUND: Depressed-type early gastric cancer (EGC) is known to repeat improvement and exacerbation of ulceration during its natural course, forming a "malignant cycle." However, it is difficult to observe the malignant cycle of EGC in clinical practice, and little is known about the clinicopathologic factors associated with this cycle. This study aimed to evaluate the malignant cycle of EGC and to determine the clinicopathologic factors associated with the time dependent morphologic change of EGC. METHODS: The medical records of EGC patients treated at the Seoul National University Hospital were retrospectively reviewed with two or more comparable endoscopic photos taken between March 1999 and December 2005. The ulcer stages in EGC were classified by the gastric ulcer stage system and evaluated for time-dependent morphologic changes according to the relevant factors. RESULTS: In this study, 231 cases of depressed-type EGC were evaluated. At the follow-up endoscopy after a median interval of 23 days, a change in ulcer stage was observed in 66 patients (29%), with improvement in 45 patients (20%) and exacerbation in 21 patients (9%). Of the 177 patients who demonstrated an active or healing stage, 45 (25%) showed improvement and 10 (6%) exhibited exacerbation. The multivariate analysis showed that the usage of antisecretory medications, mucosal cancer, and a longer interval between the two endoscopic examinations were the factors that had a significant association with improvement of the ulcer stage. CONCLUSIONS: The malignant cycle was frequently observed in depressed-type EGC. The improvement of ulceration in EGC was time dependent and associated with the usage of antisecretory medication and the depth of invasion. PMID- 19296171 TI - Trimodal imaging-assisted endoscopic mucosal resection of early Barrett's neoplasia. AB - BACKGROUND: Chromoendoscopy is traditionally used to identify inconspicuous Barrett's neoplasia and mark the boundaries before endoscopic mucosal resection (EMR). Trimodal imaging endoscopy is useful in identifying early neoplasia in Barrett's esophagus. The purpose of this study was to determine the efficacy (lateral and deep margin clearance) of trimodal imaging endoscopy-assisted EMR in early Barrett's neoplasia in a tertiary referral setting. METHODS: The entire Barrett's segment was visualized by high-resolution endoscopy followed by autofluorescence imaging, and suspicious areas were identified. Narrow band imaging and magnification was used to confirm the suspicious areas. The outer boundary of the lesion was then marked using the tip of a snare diathermy and EMR was performed using multiband mucosectomy technique by a single operator (KR). RESULTS: Sixteen patients were included: 13 patients had high-grade intraepithelial neoplasia (HGIN); 3 patients had intramucosal carcinoma (IMC); (8 males, median age, 69.5 (range, 50-77) years with Barrett's esophagus (interquartile range (IQR), 2-5 cm; median length, 3 cm). All lesions were successfully identified, using trimodal imaging endoscopy. Overall EMR was complete in 14 of 16 (81.2%) patients with early Barrett's neoplasia. Four of 16 patients (18%; all with pre-EMR HGIN) were considered to have incomplete EMR (deep margins involved), but operative histology showed only Barrett's metaplasia in 2 (histological false positive) and IMC in 1. Thus, overall 14 of 16 (87.5%) patients had complete EMR of the original lesions. In patients who did not undergo esophagectomy, there was no disease recurrence on endoscopic biopsies at a mean follow-up of 8 (IQR, 6-12) months. Three of the 16 patients (18.75%) with post-EMR bleeding were successfully treated with APC/hemoclips. CONCLUSIONS: Trimodal imaging endoscopy is a feasible alternative to chromoendoscopy to identify inconspicuous neoplasia and assist EMR of early neoplasia in Barrett's esophagus. PMID- 19296172 TI - Advanced gastric cancer: is laparoscopic gastrectomy safe? PMID- 19296173 TI - Laparoscopic resection of left liver segments using the intrahepatic Glissonian approach. AB - BACKGROUND: Recent advances in laparoscopic techniques have resulted in growing indications for laparoscopic hepatectomy. However, this procedure has not been widely developed, and anatomic segmental liver resection is not currently performed due to difficulty controlling the segmental Glissonian pedicles laparoscopically. This study aimed to report a novel technique for laparoscopic anatomic resection of left liver segments using the intrahepatic Glissonian approach based on small incisions according to anatomic landmarks such as Arantius' and round ligaments. METHODS: Nine consecutive patients underwent laparoscopic liver resection using the intrahepatic Glissonian technique from April 2007 to June 2008. Five patients underwent laparoscopic bisegmentectomy 2 3, one laparoscopic left hemihepatectomy, two resections of segment 3, and one resection of segment 4. RESULTS: One patient required a blood transfusion. The mean operation time was 180 min (range, 120-300 min), and the median hospital stay was 3 days (range, 1-5 days). No patient had postoperative signs of liver failure or bile leakage. No postoperative mortality was observed. CONCLUSION: The main advantage of the intrahepatic Glissonian procedure over other techniques is the possibility of gaining a rapid and precise access to the left Glissonian sheaths facilitating left hemihepatectomy, bisegmentectomy 2-3, and individual resections of segments 2, 3, and 4. The authors believe that the intrahepatic Glissonian technique facilitates laparoscopic liver resection and may increase the development of segment-based laparoscopic liver resection. PMID- 19296174 TI - Thermal spread and heat absorbance differences between open and laparoscopic surgeries during energized dissections by electrosurgical instruments. AB - BACKGROUND: The reported literature suggests a higher incidence of collateral damage during energized dissections in laparoscopic surgery than in open surgery, probably because the ambient environment of the two approaches causes different heat absorbance and thermal spread during energized dissection. METHODS: The experimental design involved randomized allocation of the surgical approach (4 open and 4 laparoscopic procedures) for eight adult pigs. A bipolar electrosurgery system (LigaSure) was used to perform 40 standardized regional dissections and procedures (5 per animal in randomized order). During these procedures, both white light and infrared thermographic imaging (3-5 mum) were recorded, and biopsy specimens were procured for histology. RESULTS: The core body temperatures were significantly different between the two approaches. The laparoscopic group had a smaller temperature drop (3.08 +/- 0.98 degrees C vs 1.9 +/- 0.72 degrees C; p = 0.03). Differences also were observed in the thermal spread between the laparoscopic and open groups during portal vein dissection with the Advance (1.7 +/- 0.2 mm vs 2.3 +/- 1.7 mm; p = 0.07). Both thermography and histology confirmed the safety of the LigaSure system in limiting thermal spread and necrosis at the fusion line. Significant degradation of the instruments began after 10 activations. The deterioration was faster and more extensive in laparoscopic surgery. CONCLUSIONS: Heat absorbance and thermal spread during bipolar electrosurgery are significantly different between open and laparoscopic surgery. Device performance degradation with repeat activations is more marked in laparoscopic surgery. PMID- 19296175 TI - Meta-analysis of laparoscopic and open distal gastrectomy for gastric carcinoma. PMID- 19296176 TI - Single-incision laparoscopic cholecystectomy using flexible endoscopy: saline infiltration gallbladder fossa dissection technique. AB - BACKGROUND: The introduction of natural orifice translumenal endoscopic surgery has led to the development of new techniques to accomplish minimally invasive procedures using flexible endoscopic instruments. This study evaluated a technique used in endoscopic mucosal resection and applied it to dissection of the gallbladder from the liver bed. METHODS: Eight patients underwent an elective transumbilical single-incision cholecystectomy using a flexible endoscope at the authors' institution from August 2007 to February 2008. An endoscopic injection needle was used to inject 20 ml of saline strategically into the gallbladder fossa. After infiltration, dissection of the gallbladder and hilum was performed with endoscopic instruments, whereas the cystic duct and artery were clipped using laparoscopic instruments. RESULTS: None of the eight patients had inadvertent perforation of the gallbladder during dissection. The technique of infiltrating the potential space between the gallbladder and the liver bed leads to a significantly improved visualization of the plane between them. CONCLUSION: The injection of saline to develop surgical planes is an effective tool in performing a cholecystectomy using flexible endoscopic instrumentation. The enhancement of this potential space improved visualization in all patients. This technique has great potential value for dissections and requires further evaluation of its effectiveness in other applications. PMID- 19296177 TI - British Columbia's fish health regulatory framework's contribution to sustainability goals related to salmon aquaculture. AB - Salmon farming is a significant contribution to the global seafood market to which the goal of sustainability is often applied. Diseases related to farms are perhaps the most contentious issues associated with sustainable salmon farming. We reviewed literature and policies in British Columbia, Canada, as well as interviewed key informants to examine how fish health regulations do or could support sustainability goals. We found four main obstacles to the development and application of a sustainability-based health management system. First, salmon farming faced the same challenges as other industries when trying to establish an operational definition of sustainability that captures all stakeholders' interests. Second, there was no program responsible for integrating the various regulations, responsible departments, and monitoring efforts to develop a comprehensive view of sustainability. Third, there was inadequate research base and social consensus on the criteria that should be used to track health outcomes for sustainability purposes. Fourth, the regulatory and management paradigm for salmon farming has been focused on diseases and pathogens as opposed to embracing a more inclusive health promotion model that includes biotic, abiotic, and social determinants of health. A transparent and inclusive participatory process that effectively links expert views with community and industry concerns should serve as the foundation for the next generation of health management regulations for salmon farming. PMID- 19296178 TI - Desmoid tumors of the anterior abdominal wall: results from a monocentric surgical experience and review of the literature. AB - BACKGROUND: Desmoid tumor, also known as aggressive fibromatosis, is a rare soft tissue tumor. For those cases localized in the anterior abdominal wall, radical resection and reconstruction with a mesh is indicated. Because the rarity of the disease, randomized trials are lacking, but in reported retrospective series, it is clear that although it is considered a benign lesion, local recurrence is not uncommon. METHODS: We analyzed the records of 14 consecutive patients (3 men, 11 women, mean age 36 years, range 25-51 years) with desmoid tumor of the anterior abdominal wall treated at the European Institute of Oncology. The surgical strategy was the same in all cases: wide surgical excision and immediate plastic reconstruction with mesh after intraoperative confirmation by frozen sections of disease-free margins of >1 cm. We considered long-term outcomes by using the European Organization for the Research and Treatment of Cancer QLQ-C30 as an instrument to evaluate the overall quality of the treatment delivered to these patients. RESULTS: No immediate postoperative complication was registered, and no patient developed recurrence after a median follow-up period of 55 months. Two women experienced mesh bulging within 1 year after the operation. The long-term mean global health status registered was 97 out of 100. CONCLUSIONS: Radical resection aided by intraoperative margin evaluation via frozen sections followed by immediate mesh reconstruction is a safe procedure and can provide definitive cure without functional limitations for patients with desmoid tumors of the anterior abdominal wall. PMID- 19296179 TI - Training residents to employ self-efficacy-enhancing interviewing techniques: randomized controlled trial of a standardized patient intervention. AB - BACKGROUND: Current interventions to enhance patient self-efficacy, a key mediator of health behavior, have limited primary care application. OBJECTIVE: To explore the effectiveness of an office-based intervention for training resident physicians to use self-efficacy-enhancing interviewing techniques (SEE IT). DESIGN: Randomized controlled trial. PARTICIPANTS: Family medicine and internal medicine resident physicians (N = 64) at an academic medical center. MEASUREMENTS: Resident use of SEE IT (a count of ten possible behaviors) was coded from audio recordings of the physician-patient portion of two standardized patient (SP) instructor training visits and two unannounced post-training SP visits, all involving common physical and mental health conditions and behavior change issues. One post-training SP visit involved health conditions similar to those experienced in training, while the other involved new conditions. RESULTS: Experimental group residents demonstrated significantly greater use of SEE IT than controls, starting after the first training visit and sustained through the final post-training visit. The mean effect of the intervention was significant [adjusted incidence rate ratio for increased use of SEE IT = 1.94 (95% confidence interval = 1.34, 2.79; p < 0.001)]. There were no significant effects of resident gender, race/ethnicity, specialty, training level, or SP health conditions. CONCLUSIONS: SP instructors can teach resident physicians to apply SEE IT during SP office visits, and the effects extend to health conditions beyond those used for training. Future studies should explore the effects of the intervention on practicing physicians, physician use of SEE IT during actual patient visits, and its influence on patient health behaviors and outcomes. PMID- 19296180 TI - Dilating the stenotic gastrojejunostomy after laparoscopic Roux-en-Y gastric bypass for morbid obesity: when things go wrong. PMID- 19296181 TI - The ancient technique of "gastrorrhaphy". AB - OBJECTIVE: The paper describes "gastrorrhaphy," deriving from the Greek words "gastir" meaning "abdomen" and "rhaphy" meaning "suturing," which was a technique used for the treatment of abdominal wounds. METHODS: The technique is described in detail in the texts of Celsus (first century A.D.) and in those of Galen (second century A.D.). Furthermore, references were found in Oribasius' texts (fourth century A.D.) and in the writings of two veterinarian doctors of the same period. We provide our drawings in order to elucidate the different techniques of suturing. RESULTS: Celsus described one method of "gastrorrhaphy" while Galen presented two different methods for this procedure. All three methods agree on the processes required: replacement of the prolapsed viscera, cleaning of the wound, and suturing. The difference in methods is in the way of suturing the wound; Celsus suggests stitches in layers. While Galen's first method refers to stitching of the peritoneum with the abdominal wall, his second method refers to stitching of similar structures, meaning peritoneum to peritoneum and abdominal wall to abdominal wall. CONCLUSIONS: Celsus' method strongly resembles stitching in layers with cross-sutures, while both of the Galenic techniques of gastrorrhaphy are versions of the full-thickness sutures used nowadays. It should be stressed out that Galen's methods of "gastrorrhaphy" were used by Andreas Vesalius and Ambroise Pare many centuries later. PMID- 19296182 TI - Surgical strategy for hepatocellular carcinoma patients with portal vein tumor thrombus based on prognostic factors. AB - RATIONALE: Surgical strategy for patients with hepatocellular carcinoma and portal vein tumor thrombus (PVTT) remains to be established. METHODS: From 1990 to 2008, 48 hepatocellular carcinoma patients with PVTT detected by preoperative imaging underwent hepatic resection, and their clinical data were retrospectively analyzed. Possible prognostic factors for survival were analyzed with postoperative survival curves, and significant factors were determined by univariate and multivariate analysis. The frequency of postoperative severe complications was investigated for each prognostic factor. RESULTS: Significant prognostic factors included patient age <60 years, serum total bilirubin (T-Bil) >0.8 mg/dl, serum aspartate aminotransferase >30 IU/L, serum alkaline phosphatase (ALP) >300 IU/L, tumor size >4 cm, PVTT in the main trunk (Vp4), and a surgical margin <1 mm by univariate analysis, and independent prognostic factors were serum T-Bil, ALP, and Vp4. No patient with Vp4 survived for more than 400 days after surgery, and frequency of postoperative severe complications in these Vp4 patients was significantly higher than in other Vp1-3 patients. CONCLUSION: Hepatic resection as a first-choice treatment should be carefully selected in patients with Vp4 unless emergent removal of the PVTT is required. PMID- 19296183 TI - Mesenteric venous thrombosis and factors associated with mortality: a statistical analysis with five-year follow-up. AB - OBJECTIVE: The objective was to study the factors associated with mortality in mesenteric venous thrombosis (MVT). METHODS: We reviewed all cases of bowel ischemia at our institute from 1984 to 2004 and identified 31 cases of MVT and compiled data concerning their demographics, risk factors, investigations, management, surgical procedures, and outcomes. Survival was analyzed for both 30 day and 5-year periods. RESULTS: Analysis of factors associated with mortality in our 31 case series revealed that 30-day mortality was strongly associated with colonic involvement in ischemia (p = .008) as well as short bowel syndrome (p = .028) and possibly failure to anti-coagulate the patient (p = .07). While 5-year mortality was strongly associated with "short bowel syndrome" as defined by small bowel remaining less than 100 cm (p = .031). Further study using a multivariate Cox proportional hazard analysis showed that mortality within the 30-day period was mainly related to colon ischemia with p value of .014 and an odds ratio of 17.4, while short-bowel syndrome was the predominated factor in the 5-year mortality analysis with a p value of .029 and an odds ratio of 5. CONCLUSION: Thirty-day mortality for MVT is strongly associated with colonic involvement as well as "short-bowel" syndrome, while anticoagulation may be protective. Five year survival was found to be strongly associated with "short-bowel" syndrome. PMID- 19296184 TI - Proteasome activity in a naive mosquito cell line infected with Wolbachia pipientis wAlbB. AB - We used Wolbachia pipientis strain wAlbB from Aedes albopictus Aa23 cells to infect clonal Ae. albopictus TK-6 cells, which are resistant to 5 bromodeoxyuridine. Infected TK-6 cells were cultured in medium containing 5 bromodeoxyuridine to select against Aa23 cells that might have persisted in the inoculum. Infected TK-6 lines retained the Wolbachia infection for 5 mo, indicating that their metabolic processes support Wolbachia growth and multiplication. To investigate early events after Wolbachia infection, we labeled infected cells with (35)S[methionine/cysteine]. Patterns of labeled proteins on sodium dodecyl sulfate gels were similar in control and infected cells, with the exception of a 29-kDa protein. Tandem mass spectrometry revealed that the 29-kDa band included alpha and beta subunits of the 26S proteasome. Independent confirmation of the up-regulation of the proteasome was established by probing Western blots with a monoclonal antibody to the proteasome-associated co-factor, ubiquitin. Wolbachia's loss of metabolic pathways for the synthesis of most amino acids and retention of pathways for their uptake and metabolism suggest that proteasome activation provides a mechanism whereby controlled degradation of intracellular host proteins would increase availability of amino acids to support establishment and maintenance of the Wolbachia infection. PMID- 19296185 TI - Adult jejunojejunal intussusception after jejunoileal bypass bariatric surgery. AB - The jejunoileal bypass is one of the bariatric surgical options which have been abandoned for two reasons: first, it leads to severe malnutrition and liver failure and, second, the bypassed jejunum-being a blind loop-is susceptible to bacterial accumulation which might become a source of sepsis due to bacterial translocation. We hereby report a case of a 27-year-old lady who presented with jejunojejunal intussusception of the blind jejunal loop 2 years after jejunoileal bypass surgery. This complication could have led to serious consequences if it was not managed in the appropriate time. PMID- 19296187 TI - Standards of reporting: the CONSORT, QUORUM, and STROBE guidelines. PMID- 19296186 TI - Improved early clinical outcomes of RP/PS mobile-bearing total knee arthroplasties. AB - The rotating-platform posterior-stabilized (RP/PS) prosthesis was developed to take advantage of the benefits of the traditional RP mobile-bearing system and the posterior-stabilized design. This nonconsecutive cohort study compared the clinical outcomes of TKAs performed using a RP/PS mobile system or a floating platform (FP) system. The clinical outcomes of 93 TKAs with a RP/PS prosthesis were compared with the same number of TKAs with a FP mobile-bearing prosthesis at 6, 12, and 24 months after surgery. Clinical outcomes differed between the FP and RP/PS groups as a function of length of followup. In the FP group, most outcomes peaked at 12 months and then deteriorated, whereas in the RP/PS group, outcomes stabilized or continued to improve between 12 and 24 months. The RP/PS group had greater maximum flexion throughout followup and better clinical outcomes 24 months after surgery. Patient satisfaction was superior in the RP/RS group. This study suggests the RP/PS prosthesis provides better functional outcomes, including greater maximum flexion and better patient satisfaction. We propose the RP/PS mobile-bearing system is a more attractive option than the FP mobile bearing system for patient populations of elderly women similar to patients enrolled in our study. LEVEL OF EVIDENCE: Level III, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19296188 TI - Simulation-based education in Canada: will anesthesia lead in the future? PMID- 19296189 TI - The Hunsaker Mon-Jet tube with jet ventilation is effective for microlaryngeal surgery. AB - PURPOSE: Microlaryngeal surgery involves a delicate balance between airway control and appropriate visualization of laryngeal anatomy. When the self centering, laser-safe Hunsaker Mon-Jet tube (Xomed, Jacksonville, FL) was introduced in 1994, to provide subglottic ventilation, the needs of both anesthesiologist and surgeon appeared to have been adequately met. However, limited data exists regarding the efficacy of this device in a large patient series. The aim of this cohort study was to explore the spectrum of patients and procedures for which this technique could be used. METHODS: We report a retrospective study of 552 patients who had undergone microlaryngeal surgery between January 1995 and June 2005, utilizing the Hunsaker Mon-Jet tube and automated jet ventilator to provide subglottic ventilation. In addition to patient demographics, overall success of ventilation and several perioperative outcomes were evaluated, including the incidence of complications. RESULTS: In combination with an automatic jet ventilator, the Hunsaker Mon-Jet tube was successfully utilized in more than 98% of the 552 patients, including those with severe co-morbidities, obesity and difficult airway access, providing optimal visualization of the vocal cords and excellent surgical access for a wide range of laryngeal disorders, including laser treatment. In less than 2% of patients, hypoxia, hypercarbia, or both, necessitated exchange of the Hunsaker Mon-Jet tube for a standard or laser-safe endotracheal tube with rapid resolution of the precipitating cause. CONCLUSIONS: Subglottic ventilation via the Hunsaker Mon-Jet tube with an automated jet ventilator may be considered an effective, safe and versatile technique for the anesthetic management of microlaryngeal surgery. PMID- 19296190 TI - Critical care services in Ontario: a survey-based assessment of current and future resource needs. AB - PURPOSE: In response to the challenges of an aging population and decreasing workforce, the provision of critical care services has been a target for quality and efficiency improvement efforts. Reliable data on available critical care resources is a necessary first step in informing these efforts. We sought to describe the availability of critical care resources, forecast the future requirement for the highest-level critical care beds and to determine the physician management models in critical care units in Ontario, Canada. METHODS: In June 2006, self-administered questionnaires were mailed to the Chief Executive Officers of all acute care hospitals, identified through the Ontario government's hospital database. The questionnaire solicited information on the number and type of critical care units, number of beds, technological resources and management of each unit. RESULTS: Responses were obtained from 174 (100%) hospitals, with 126 (73%) reporting one or more critical care units. We identified 213 critical care units in the province, representing 1789 critical care beds. Over half (59%) of these beds provided mechanical ventilation on a regular basis, representing a capacity of 14.9 critical care and 8.7 mechanically ventilated beds per 100,000 population. Sixty-three percent of units with capacity for mechanical ventilation involved an intensivist in admission and coordination of care. Based on current utilization, the demand for mechanically ventilated beds by 2026 is forecast to increase by 57% over levels available in 2006. Assuming 80% bed utilization, it is estimated that an additional 810 ventilated beds will be needed by 2026. CONCLUSION: Current utilization suggests a substantial increase in the need for the highest-level critical care beds over the next two decades. Our findings also indicate that non-intensivists direct care decisions in a large number of responding units. Unless major investments are made, significant improvements in efficiency will be required to maintain future access to these services. PMID- 19296191 TI - Direct effect of ropivacaine involves lipoxygenase pathway activation in rat aortic smooth muscle. AB - PURPOSE: Ropivacaine is a long-acting amino-amide local anesthetic that induces vasoconstriction in vitro and in vivo. The aim of this study was to investigate the pathways involved in arachidonic acid metabolism associated with S ropivacaine-induced contraction of rat aortic smooth muscle in vitro. METHODS: Rat thoracic aortic rings without endothelium were isolated and suspended for isometric tension recording. Cumulative dose-response curves were generated with concentrations of 10(-5) to 10(-3) M ropivacaine enantiomer in the presence or absence of quinacrine dihydrochloride, nordihydroguaiaretic acid, quinacrine dihydrochloride plus nordihydroguaiaretic acid, indomethacin, fluconazole, AA 861, and verapamil. The maximal S-ropivacaine-induced contractile response achieved at 3x10(-4) M was also assessed in aortic rings pretreated with normal or calcium-free Krebs solution. RESULTS: Ropivacaine enantiomers induced dose dependent biphasic contractions in aortic rings. S-ropivacaine (10(-4), 3x10(-4) M) induced a stronger contraction than R-ropivacaine. Quinacrine dihydrochloride (2x10(-5), 4x10(-5) M) attenuated the S-ropivacaine-induced biphasic contraction in a dose-dependent manner. Indomethacin (3x10(-5), 6x10(-5) M), nordihydroguaiaretic acid (10(-5) M), and AA-861 (10(-5) M) also attenuated the S ropivacaine-induced dose-dependent biphasic contraction, whereas fluconazole (3x10(-5)) had no effect. Combined pretreatment with quinacrine dihydrochloride and nordihydroguaiaretic acid almost completely abolished the S-ropivacaine induced contraction. S-ropivacaine-induced contractile responses were attenuated by verapamil (10(-5) M) and calcium-free Krebs solution. CONCLUSION: S ropivacaine induces dose-dependent biphasic contractions in rat aortic smooth muscle through a mechanism requiring extracellular calcium that is mediated by activation of the lipoxygenase pathway and, to a lesser extent, the cyclooxygenase pathway. PMID- 19296192 TI - Tetanus in developing countries: a case series and review. AB - PURPOSE: Few anesthesiologists have expertise in the diagnosis and treatment of tetanus, a disease that remains prevalent in developing countries. We report on a series of four cases of tetanus cases recently encountered in Rwanda. We review the clinical epidemiology, pathophysiology, diagnosis and the treatment of tetanus, and provide implications for anesthesiologists and critical care physicians. CLINICAL FEATURES: We report four cases, two involving adults who were inadequately vaccinated and experienced injuries, and two involving neonates, both of whom underwent umbilical cord transection using unsterilized equipment. All patients required tracheal intubation, and were mechanically ventilated when equipment was available. One adult and one neonate succumbed to the disease. These cases highlight the difficulties of diagnosis and management of complicated diseases in the resource-challenged health care setting of developing countries. CONCLUSIONS: The differential diagnosis of tetanus may be confusing, and survival depends on the rapidity of treatment with antitoxin, as well as adequate supportive care. High doses of sedatives and muscle relaxants, as well as prolonged mechanical ventilation, are usually necessary. Mortality remains high, usually resulting from late respiratory failure and cardiovascular collapse, associated with autonomic instability. Anesthesiologists and critical care physicians have an important role to play in the management of these patients. Increased involvement in humanitarian health organizations, immigration from developing countries, and emergence of high risk groups in developed countries will likely result in more exposure of anesthesiologists to the complexities of this disease. PMID- 19296193 TI - Non-cardiac implantable electrical devices: brief review and implications for anesthesiologists. AB - PURPOSE: Patients with implanted electrical devices (IEDs) are vulnerable to electromagnetic interference (EMI) from electrical equipment used in the perioperative environment. As the indications for non-cardiac IEDs increase, so too does the likelihood of anesthesiologists encountering such patients who present for surgery. This article reviews the important anesthetic considerations for patients with implanted non-cardiac electrical devices. SOURCE: We searched Medline (1980 to December 2008) and Embase (1980 to December 2008) and reviewed all relevant manuscripts regarding non-cardiac IEDs and anesthesia. PRINCIPAL FINDINGS: There are numerous non-cardiac IEDs currently in use, including deep brain stimulators, vagal nerve stimulators, gastric stimulators, phrenic nerve stimulators, sacral nerve stimulators, and bone stimulators. A key goal in the perioperative management of these patients is to reduce the risk of adverse events resulting from device interactions with electrocautery, external emergency cardiac defibrillation, peripheral nerve stimulation, neuraxial anesthesia, and magnetic resonance imaging. We offer a general management strategy for patients with IEDs presenting for surgery. CONCLUSIONS: In the absence of available anesthesia management guidelines regarding non-cardiac IEDs in the perioperative setting, anesthesiologists should appreciate the basic principles pertaining to IEDs to ensure appropriate risk reduction strategies in order to enhance patient safety. PMID- 19296194 TI - Perioperative subepicardial aneurysm (impending myocardial rupture) after drug eluting stent thrombosis. PMID- 19296196 TI - Low recurrence rate of a two-layered closure repair for primary and recurrent midline incisional hernia without mesh. AB - BACKGROUND: Incisional hernia is a serious complication after abdominal surgery and occurs in 11-23% of laparotomies. Repair can be done, for instance, with a direct suture technique, but recurrence rates are high. Recent literature advises the use of mesh repair. In contrast to this development, we studied the use of a direct suture repair in a separate layer technique. The objective of this retrospective observational study is to assess the outcomes (recurrences and complications) of a two-layered open closure repair for primary and recurrent midline incisional hernia without the use of mesh. METHODS: In an observational retrospective cohort study, we analysed the hospital and outpatient records of 77 consecutive patients who underwent surgery for a primary or recurrent incisional hernia between 1st May 2002 and 8th November 2006. The repair consisted of separate continuous suturing of the anterior and posterior fascia, including the rectus muscle, after extensive intra-abdominal adhesiolysis. RESULTS: Forty-one men (53.2%) and 36 women (46.8%) underwent surgery. Sixty-three operations (81.8%) were primary repairs and 14 (18.2%) were repairs for a recurrent incisional hernia. Of the 66 patients, on physical examination, three had a recurrence (4.5%) after an average follow-up of 2.6 years. The 30-day postoperative mortality was 1.1%. Wound infection was seen in five patients (6.5%). CONCLUSIONS: A two-layered suture repair for primary and recurrent incisional hernia repair without mesh with extensive adhesiolysis was associated with a recurrence rate comparable to mesh repair and had an acceptable complication rate. PMID- 19296197 TI - A novel endo-glucanase from the thermophilic bacterium Geobacillus sp. 70PC53 with high activity and stability over a broad range of temperatures. AB - A thermophilic Geobacillus bacterium secreting high activity of endo-glucanase (EC 3.2.1.4) was isolated from rice straw compost supplemented with pig manure. A full-length gene of 1,104 bp, celA, encoding this glycosyl hydrolase family 5 endo-glucanase of 368 amino acids was isolated. No related gene from Geobacillus has been reported previously. The recombinant CelA expressed in Escherichia coli had an optimal activity at 65 degrees C and pH 5.0, and it exhibited tenfold greater specific activity than the commercially available Trichoderma reesei endo glucanase. CelA displayed activity over a broad temperature range from 45 to 75 degrees C and was a thermostable enzyme with 90% activity retained after heating at 65 degrees C for 6 h. Interestingly, CelA activity could be enhanced by 100% in the presence of 2 mM MnSO(4). CelA had high specific activity over beta-D glucan from barley and Lichenan, making it a potentially useful enzyme in biofuel and food industries. PMID- 19296198 TI - Buccal dental microwear variability in extant African Hominoidea: taxonomy versus ecology. AB - Buccal microwear patterns on teeth are good indicators of the abrasiveness of foodstuffs and have been used to trace the dietary habits of fossil species, including primates and hominids. However, few studies have addressed the variability of this microwear. The abrasiveness of dietary components depends not only on the hardness of the particles ingested, but also on the presence of dust and other exogenous elements introduced during food processing. These elements are responsible for the microwear typology observed on the enamel surfaces of primate teeth. Here we analyzed the variability of buccal microwear patterns in African Great Apes (Gorilla gorilla and Pan troglodytes), using tooth molds obtained from the original specimens held in several osteological collections. Our results suggest that ecological adaptations at subspecies or population level account for differences in microwear patterns, which are attributed to habitat and ecological conditions within populations rather than differences between species. The findings from studies on the variability of buccal dental microwear in extant species will contribute to a better understanding of extinct hominids' diet and ecology. PMID- 19296199 TI - A retrospective analysis of bortezomib therapy for Japanese patients with relapsed or refractory multiple myeloma: beta2-microglobulin associated with time to progression. AB - Bortezomib is approved for the treatment of patients with relapsed or refractory multiple myeloma (MM), but only a few clinical studies for Japanese patients who were treated with bortezomib have been reported. We retrospectively analyzed 40 patients with relapsed or refractory MM who have received bortezomib at three collaborating centers in Miyagi prefecture in Japan. All the patients have been received bortezomib in combination with dexamethasone. Responses were determined using International Myeloma Working Group uniform response criteria. The overall response was observed in 30 patients (75%), including very good partial response in 8 patients (20%), and partial response in 22 patients (55%). The median time to disease progression was 8.7 months, and the median overall survival has not been reached. The factors affecting time to disease progression were International Staging System stage, serum beta2-microglobulin level, and number of treatment cycles. The most common grade 3 and 4 adverse events were thrombocytopenia (50%), peripheral neuropathy (25%), leukopenia (25%), and herpes zoster infection (25%). Thus, bortezomib is well tolerated and effective for Japanese patients with relapsed or refractory MM. Our results suggest that serum beta2-microglobulin level may be a marker of prognosis on bortezomib therapy for patients with relapsed or refractory MM although further studies are needed. PMID- 19296200 TI - A new beginning for Anatomical Science International (ASI) with Springer: a message from the editor-in-chief. PMID- 19296201 TI - Complete genomic characterization of a European type 1 porcine reproductive and respiratory syndrome virus isolate in Korea. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) isolates belonging to the European genotype 1 have recently emerged in South Korea, suggesting potential problems for disease control. In the present study, we attempted to determine the complete nucleotide sequence of the first Korean type 1 PRRSV isolate, designated KNU-07. The full-length genome of KNU-07 was found to be 15,038 nucleotides in length, which was 60 nucleotides shorter than the type 1 prototype strain Lelystad due to a notable 60-bp deletion within the nonstructural protein 2 (NSP2). The KNU-07 genome was shown to consist of a 221 nucleotide (nt) 5' untranslated region (UTR), a 14,703-nt protein-coding region, and a 114-nt 3' UTR, followed by a 42-73-bp poly(A) tail. A nucleotide sequence comparison of the KNU-07 genome with 20 complete PRRSV genomes revealed a 10.5 13.3% and 39.5-40.3% divergence from type 1 and type 2 strains, respectively, at the genome level, indicating a high similarity to the virus strains commonly identified as the European genotype. In order to investigate genetic variation and to understand the molecular evolution of the type 1 isolate in Korea, extensive phylogenetic analyses were performed using the ORF5 and ORF7 nucleotide sequences of published type 1 PRRSV isolates. The data further indicated that the newly emerging type 1 isolate KNU-07 belongs to the recently proposed pan European subtype 1. Taken together, the results of this study describe the genomic characterization of the type 1 PRRSV isolated in South Korea, suggesting a recent introduction of the virus typical for this genotype that has commonly appeared worldwide. PMID- 19296202 TI - Physiological, biochemical, and ultrastructural responses of the green macroalga Urospora penicilliformis from Arctic Spitsbergen to UV radiation. AB - Exposure of the filamentous turf green alga Urospora penicilliformis to ambient and artificial ultraviolet radiation (UVR) revealed a considerable resilient species. This explains the ability of this alga to thrive in the middle-upper intertidal zones of the Arctic sea where it is periodically exposed to environmental extremes. A transient UVR effect on photosynthesis under photosynthetically active radiation (PAR) + UV-A and PAR + UV-A + UV-B was found, but dynamic recovery of photoinhibition was observed immediately after reduction of the photon fluence rate of PAR in the absence or presence of background UVR under laboratory and natural solar radiation, respectively. Chlorophylls, carotenoids, and xanthophyll cycle pigments (violaxanthin, antheraxanthin, and zeaxanthin) concentrations were not significantly different between freshly collected samples and filaments exposed to additional laboratory radiation treatment. The ultrastructure of the U. penicilliformis gametophytes showed that the cells are well adapted to UVR. No significant ultrastructural alterations were observed in filaments exposed to different spectral irradiance in the laboratory compared to in situ acclimated specimen. The antioxidant alpha tocopherol was detected in minute quantity while the search for flavonoid-like compounds was negative. Other UV screening strategies or certain genetically fixed physiological protective mechanism could be operating in this species responsible for their occurrence in higher shoreline and ecological success. Further molecular and biochemical studies are needed to elucidate the stress resistance in this turf alga. There is an indication that the extremely thick cell wall of U. penicilliformis gametophytes covered with mucilage sheath and dense layer of mineral depositions may provide a shield against unfavorable environmental conditions in general and against UVR in particular. PMID- 19296203 TI - Management of distal tibial intra-articular fractures with circular external fixation. AB - The treatment of tibial plafond fractures requires careful management of the soft tissue envelope, reconstruction of the articular surface and stable fixation with minimal additional damage. Thirty cases of AO type 43 C tibial fractures were treated by transosseous osteosynthesis (Ilizarov technique). The external fixator constructs used were Ilizarov (Transosseous osteosynthesis: theoretical and clinical aspects of the regeneration and growth of tissue, Springer, Berlin, 1992) and Sheffield (Classification AO des fractures, Springer, Berlin, 1987) circular fixator systems. All tibial plafond fractures healed. Using radiological criteria for assessment of reduction of the articular fragments and the clinical scoring system described by Teeny and Wiss, there were excellent and good restoration of articular structure in 27 cases and good clinical results in 14. This treatment method compares well with previous published series and is to be recommended for this group of difficult fractures. PMID- 19296204 TI - Is catheter-related thrombosis after cardiac surgery clinically relevant? PMID- 19296205 TI - Rotavirus and coxsackievirus infection activated different profiles of toll-like receptors and chemokines in intestinal epithelial cells. AB - OBJECTIVE: To understand the inflammatory-immune response in intestinal epithelial cells after infection of rotavirus and coxsackievirus B3. METHODS: We examined by quantitative PCR the expression profiles of genes encoding five toll like receptors (TLR) and levels of three chemokines in response to rotavirus and coxsackievirus B3 infection in a human intestinal epithelial cell line (HT-29 cells). RESULTS: We demonstrated that rotavirus induced significantly increased levels of mRNA expression for TLR2, TLR3, TLR7 and TLR8 in HT-29 cells in a time dependent manner. In contrast, coxsackievirus B3 did not stimulate mRNA expression for TLR3. Rotavirus and coxsackievirus B3 also induced higher levels of mRNA expression for RANTES, IP-10 and IL-8 during the period of infection in a different manner. Finally, significantly elevated levels of RANTES, IP-10 and IL 8 were detected by ELISA in rotavirus-infected cells from 24 to 48 h. CONCLUSION: Our findings suggest that different patterns of TLRs and chemokines were induced in the initiation and modulation of immune response to rotavirus and coxsackievirus B3 infection. PMID- 19296206 TI - Investigation of effects of Lacidipine, Ramipril and Valsartan on DNA damage and oxidative stress occurred in acute and chronic periods following isoproterenol induced myocardial infarct in rats. AB - In this study, effects of Lacidipine (LAC), Ramipril (RAM) and Valsartan (VAL) on DNA damage and oxidative stress occurred in acute and chronic periods after isoproterenol (ISO)-induced myocardial infarct (MI) were investigated in rats. LAC, RAM and VAL had been administered by oral gavage at 3, 3 and 30 mg/kg doses, respectively, in acute and chronic periods following MI. In acute MI model, LAC, RAM and VAL had been administered once per day to rat groups during 30 days. On days 29 and 30, the rats of the acute MI control and drug treatment groups were administered 180 mg/kg ISO, subcutaneously at an interval of 24 h. In chronic MI model, LAC, RAM and VAL had been administered to rat groups during 30 days, and on the 1st and 2nd days, the rats of the chronic MI control and drug treatment groups were administered ISO, by the same way. After this period, routine biochemistry indicators of MI, alanin aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine kinase (CK), creatine kinase-isoenzymes (CK-MB), troponin I (TnI) and nitric oxide (NO), oxidative stress indicator, has been measured in the serums obtained from rat's blood. Also, 7,8-Dihydro-8-oxo-guanine (8-OHGua), which is an indicator of DNA damage level, has been determined in whole blood. After MI diagnosis, the relationships among the 8-OHGua, NO and clinic MI indicators have been determined. Results have been evaluated by comparing with that of control group. In control groups, the clinic MI indicators have been found to be statistically higher than the drug groups. In parallel to this increase in MI indicators, there have been determined a significant decrease in NO levels and an increase in 8 OHGua level. There was no significant difference in the rat groups which received drugs without MI induction. We have observed that the level of 8-OHGua which increased after MI in both acute and chronic periods decreased by LAC, RAM and VAL when compared to acute and chronic MI control groups. In conclusion, it has been determined that oxidative stress has been increased after ISO induced MI model and this stress reduces NO and even damages DNA. LAC, RAM and VAL may decrease the severity of MI and prevent DNA damage by reducing oxidative stress. PMID- 19296207 TI - Segmented biopolyurethanes for medical applications. AB - Polyurethanes are one of the most popular groups of biomaterials applied for medical devices. Their segmented block copolymeric character endows them a wide range of versatility in terms of tailoring their physical properties, blood and tissue compatibility. Polyester- and polyether-urethanes have been modified with hydroxypropyl cellulose aiming the change of their surface and bulk characteristics to confer them biomaterial qualities. In this respect, dynamic contact angle measurements, dynamic mechanical analyses accompanied by mechanical testing have been done. Platelet adhesion test has been carried out in vitro and the use of hydroxypropyl cellulose in the polyurethane matrix reduces the platelet adhesion and therefore recommends them as candidates for biocompatible materials. PMID- 19296208 TI - Clinical study shows improved absorption of desmopressin with novel formulation. AB - PURPOSE: To create improved pharmaceutical formulations for nasal and sublingual administration of desmopressin and investigate their pharmacokinetic profiles in comparison with a commercial nasal liquid spray and finally to evaluate the volunteers' opinions on the different dosage forms. METHODS: Both formulations were based on the characteristics of interactive mixtures. The nasal powder spray was produced by a rotary evaporator technique with sodium starch glycolate as carrier material and the sublingual tablet by direct compression after dry mixing with mannitol as carrier. The clinical study was an open-label, randomised cross over pharmacokinetic study in healthy volunteers. RESULTS: The nasal powder formulation gave a threefold increase in the absorption, unaltered time to maximum plasma concentration and a tendency to lower variability in the amount absorbed compared with the liquid spray. The powder was reported to be more irritating than the liquid but was still well accepted by the volunteers. The tablet did not improve the uptake of desmopressin, likely because of a poor disintegration sublingually. CONCLUSIONS: The nasal powder formulation is a promising new dosage form for the delivery of desmopressin and other compounds. The sublingual tablet has a beneficial means of production and may be further developed by decreasing its disintegration time. PMID- 19296209 TI - The flavonoid quercetin induces changes in mitochondrial permeability by inhibiting adenine nucleotide translocase. AB - This study shows the effects of the flavonoid quercetin on diverse mitochondrial functions, among them membrane permeability. Our findings indicate that the addition of 50 microM quercetin did not produce reactive oxygen derived species; however, it inhibited the oxidative stress induced after the addition of Fe(2)/H(2)O(2) by about 38%. At this concentration, quercetin also promoted a fast calcium release, inhibited oxidative phosphorylation, stimulated oxygen consumption, and decreased membrane potential. In addition 50 microM quercetin inhibited the adenine nucleotide translocase (ANT) by 46%. These effects induced the opening of the permeability transition pore and release of cytochrome c, by its interaction with a component of the non-specific pore complex, fixed to the carrier in the conformation c, as carboxyatractyloside does. Quercetin-induced permeability transition pore opening was inhibited by 0.5 microM cyclosporin A, but, interestingly, the release of cytochrome c was not inhibited by the immunosuppressor, as quercetin was found to disrupt the outer membrane. PMID- 19296210 TI - Effects of 1,4-butanediol administration on oxidative stress in rat brain: study of the neurotoxicity of gamma-hydroxybutyric acid in vivo. AB - gamma-Hydroxybutyric acid (GHB) is a naturally occurring compound in the central nervous system (CNS) whose tissue concentration are highly increased in the neurometabolic-inherited deficiency of succinic semialdehyde dehydrogenase (SSADH) activity or due to intoxication. SSADH deficiency is biochemically characterized by increased concentrations of GHB in tissues, cerebrospinal fluid, blood and urine of affected patients. Clinical manifestations are variable and include retardation of mental, motor, and language development along with other neurological symptoms, such as hypotonia, ataxia and seizures, whose underlying mechanisms are practically unknown. The precursor of GHB, 1,4-butanediol (1,4-BD) has been used to study the mechanisms of in vivo GHB neurotoxicity. Therefore, in the present work, the effect of acute administration of 20 or 120 mg/Kg 1,4-BD was investigated on various parameters of oxidative stress, such as spontaneous chemiluminescence, thiobarbituric acid-reactive substances (TBA-RS), total antioxidant reactivity (TAR), sulfhydryl and protein carbonyl contents, as well as the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in homogenates from cerebral cortex of 14 day-old Wistar rats. Acute administration of 120 mg/Kg 1,4-BD significantly increased spontaneous chemiluminescence and TBA-RS levels, while TAR measurement was markedly diminished, whereas injection of a lower dose (20 mg/Kg) did not change the parameters examined. Other parameters of oxidative stress evaluated were not affected by administration of 1,4-BD. These results indicate that 1,4-BD induces in vivo oxidative stress by stimulating lipid peroxidation and decreasing the non-enzymatic antioxidant defenses in cerebral cortex of young rats. If these effects also occur in humans, it is possible that they might contribute to the brain damage found in SSADH-deficient patients and possibly in individuals intoxicated by GHB or its prodrugs (gamma-butyrolactone or 1,4-BD). PMID- 19296211 TI - Effects of adult-onset streptozotocin-induced diabetes on the rat brain antioxidant status and the activities of acetylcholinesterase, (Na(+),K (+))- and Mg(2+)-ATPase: modulation by L-cysteine. AB - Uncontrolled diabetes is known to affect the nervous system. The aim of this study was to investigate the effect of the antioxidant L: -cysteine (Cys) on the changes caused by adult-onset streptozotocin (STZ)-induced diabetes on the rat brain total antioxidant status (TAS) and the activities of acetylcholinesterase (AChE), (Na(+),K(+))-ATPase and Mg(2+)-ATPase. Thirty-eight male Wistar rats were divided into six groups: C(A) (8-week-control), C(B) (8-week-control + 1-week saline-treated), C + Cys (8-week-control + 1-week-Cys-treated), D(A) (8-week diabetic), D(B) (8-week-diabetic + 1-week-saline-treated) and D + Cys (8-week diabetic + 1-week-Cys-treated). All diabetic rats were once treated with an intraperitoneal (i.p.) STZ injection (50 mg/kg body weight) at the beginning of the experiment, while all Cys-treated groups received i.p. injections of Cys 7 mg/kg body weight (daily, for 1-week, during the 9th-week). Whole rat brain parameters were measured spectrophotometrically. In vitro incubation with 0.83 mM of Cys or 10 mM of STZ for 3 h was performed on brain homogenate samples from groups C(B) and D(B), in order to study the enzymes' activities. Diabetic rats exhibited a statistically significant reduction in brain TAS (-28%, D(A) vs C(A); 30%, D(B) vs C(B)) that was reversed after 1-week-Cys-administration into basal levels. Diabetes caused a significant increase in AChE activity (+27%, D(A) vs C(A); +15%, D(B) vs C(B)), that was further enhanced by Cys-administration (+57%, D + Cys vs C(B)). The C + Cys group exhibited no significant difference compared to the C(B) group in TAS (+2%), but showed a significantly increased AChE activity (+66%, C + Cys vs C(B)). Diabetic rats exhibited a significant reduction in the activity of Na(+),K(+)-ATPase (-36%, D(A) vs C(A);-48%, D(B) vs C(B)) that was not reversed after 1-week Cys administration. However, in vitro incubation with Cys partially reversed the diabetes-induced Na(+),K(+)-ATPase inhibition. Mg(2+)-ATPase activity was not affected by STZ-induced diabetes, while Cys caused a significant inhibition of the enzyme, both in vivo (-14%, C + Cys vs C(B);-17%, D + Cys vs C(B)) and in vitro (-16%, D(B) + in vitro Cys vs C(B)). In vitro incubation with STZ had no effect on the studied enzymes. The present data revealed a protective role for Cys towards the oxidative effect of diabetes on the adult rat brain. Moreover, an increase in whole brain AChE activity due to diabetes was recorded (not repeatedly established in the literature, since contradictory findings exist), that was further increased by Cys. The inhibition of Na(+),K(+)-ATPase reflects a possible mechanism through which untreated diabetes could affect neuronal excitability, metabolic energy production and certain systems of neurotransmission. As concerns the use of Cys as a neuroprotective agent against diabetes, our in vitro findings could be indicative of a possible protective role of Cys under different in vivo experimental conditions. PMID- 19296212 TI - Illusory memories of emotionally charged words in autism spectrum disorder: further evidence for atypical emotion processing outside the social domain. AB - Recent evidence suggests that individuals with ASD may not accumulate distinct representations of emotional information throughout development. On the basis of this observation we predicted that such individuals would not be any less likely to falsely remember emotionally significant as compared to neutral words when such illusory memories are induced by asking participants to study lists of words that are orthographically associated to these words. Our findings showed that typical participants are far less likely to experience illusory memories of emotionally charged as compared to neutral words. Individuals with ASD, on the other hand, did not exhibit this emotional modulation of false memories. We discuss this finding in relation to the role of emotional processing atypicalities in ASD. PMID- 19296213 TI - Heritability of high reading ability and its interaction with parental education. AB - Moderation of the level of genetic influence on children's high reading ability by environmental influences associated with parental education was explored in two independent samples of identical and fraternal twins from the United States and Great Britain. For both samples, the heritability of high reading performance increased significantly with lower levels of parental education. Thus, resilience (high reading ability despite lower environmental support) is more strongly influenced by genotype than is high reading ability with higher environmental support. This result provides a coherent account when considered alongside results of previous research showing that heritability for low reading ability decreased with lower levels of parental education. PMID- 19296214 TI - Further validation of a measure of proactive and reactive aggression within a clinical child population. AB - The current study provided further validity for the use of a measure of proactive and reactive aggression that was first developed by Little and his colleagues. Confirmatory factor analyses performed with a high-risk clinical population (N = 107) revealed that the model was a good fit to the data. Structural equation models revealed significant associations in the expected direction between the Little et al. measure and another measure of proactive and reactive aggression developed by Dodge and Coie. Further analyses revealed that the proactive aggression subscales of the Little et al. and Dodge and Coie measures were significantly related to conduct problems. However, the reactive aggression subscale of the Little et al. measure was not related to social problems while both the proactive and reactive aggression subscales of the Dodge and Coie measure were. PMID- 19296215 TI - No evidence of familial correlation in breast cancer metastasis. AB - Animal experiments support the hypothesis that the metastatic potential of breast cancer is a heritable trait of the host. Our objective was to evaluate correlations in metastasis occurrence in large families with multiple cases of breast cancer. We evaluated correlation among pairs of relatives in the occurrence and timing of distant metastasis using retrospective cohort data from 743 female breast cancer patients in 242 families. We adjusted for correlation in their age at diagnosis, year of diagnosis, educational level, lymph node involvement, and estrogen receptor status. Distant metastasis occurred in 255 patients (34.3%) during mean followup of 11.7 years. None of the correlation coefficients for metastasis in blood relatives differed significantly from zero. The estimated correlation coefficient in first-degree relatives was -0.03 (95% confidence interval -0.11 to 0.06). These findings suggest that a family history of metastatic breast cancer does not contribute substantially to risk of metastasis for breast cancer patients. PMID- 19296217 TI - Dimethylarginine levels in cerebrospinal fluid of hyperacute ischemic stroke patients are associated with stroke severity. AB - We hypothesise that asymmetric and symmetric dimethylarginine (ADMA, SDMA) are released in cerebrospinal fluid (CSF) due to ischemia-induced proteolysis and that CSF dimethylarginines are related to stroke severity. ADMA and SDMA were measured in CSF of 88 patients with ischemic stroke or TIA within 24 h after stroke onset (mean 8.6 h) and in 24 controls. Stroke severity was assessed by the National Institutes of Health Stroke Scale (NIHSS) score at admission. Outcome was evaluated by institutionalization due to stroke and the modified Rankin scale. Dimethylarginine levels were higher in patients with stroke than in TIA patients, who had higher levels than controls and correlated with the NIHSS. Logistic regression analysis confirmed that dimethylarginines were independently associated with stroke severity. The SDMA/ADMA ratio did not differ significantly between controls and stroke patients. CSF dimethylarginine levels are increased in hyperacute ischemic stroke and are associated with stroke severity. PMID- 19296218 TI - Relationship between caregiver hopefulness and satisfaction with their children's mental health services. AB - Relationships among the Youth Services Survey for Families (YSSF) Indicators, selected demographic and clinical variables, and caregiver-rated hopefulness and children's problem severity were examined in a random sample of 1,500 caregivers of Medicaid-enrolled youth in a satisfaction study with Kentucky's 14 community mental health centers. Multiple linear regressions revealed that caregiver services and caregiver hopefulness were excellent predictors of two and three YSSF Indicators, respectively. Providers need to respond to caregiver concerns when developing support services and understand the mechanisms associated with these services' effectiveness. Tracking changes in caregiver and youth hopefulness during treatment may improve services and retention and bridge the gap between research and the important construct of hope. PMID- 19296219 TI - Regeneration of graft livers and limited contribution of extrahepatic cells after partial liver transplantation in humans. AB - BACKGROUND: Liver regeneration is still not fully understood. Partial liver transplantation (LT) can provide the opportunity to investigate the mechanisms of liver regeneration, including the contribution of extrahepatic cells to liver regeneration. METHODS: Of 61 patients transplanted with partial liver graft between August 1997 and October 2006, 56 patients were studied, including 49 adults and 7 children. Sequential computed tomography volumetric analysis was performed for volume measurement, while proliferating cell nuclear antigen (PCNA) labeling index was investigated for liver cell proliferation in nonprotocol liver biopsy specimens. In addition, 15 male recipients who had female liver grafts were investigated in order to detect Y chromosomes as extrahepatic cells in nonprotocol liver biopsy specimens. RESULTS: Graft volume per standard liver volume was markedly increased after adult-to-adult living-donor (LD) LT. In pediatric transplants, there was no volume increase over time. PCNA labeling index was vigorous in adult-to-adult LDLT in the early period after LDLT. No Y chromosome was evident in hepatocytes from female-donor male-recipient grafts during or after liver regeneration. However, in the cases of failing grafts of this type, many Y-chromosome-positive cells were observed in the graft liver. The character of those cells was CD34(-), CK9(-), hepatocyte-specific antigen(-), and CD68(+/-). CONCLUSION: In adult-to-adult LDLT, vigorous liver regeneration occurs in the graft liver, demonstrated by not only volumetric but cell kinetic analysis. Involvement of extrahepatic cells in normal liver regeneration seems limited. PMID- 19296220 TI - Correlates of smoking cessation among Filipino immigrant men. AB - A survey on tobacco use among 318 Filipino immigrant men aged 40-75 years was conducted in Los Angeles, California. Those who reported more English language use with their family, friends and neighbors (OR = 1.31) and who lived in households with complete smoking prohibition (OR = 3.82) were more likely to be successful in quitting smoking. Those who endorsed more positive beliefs on physical and social consequences of smoking (OR = 0.69) and who had mostly smoking friends (OR = 0.37) were less likely to be successful in quitting smoking. Our findings suggest that prohibiting smoking in households, creating social networks of non-smokers, and education or counseling are important components of a smoking cessation intervention for Filipino immigrant men. PMID- 19296221 TI - Experimental-computational evaluation of human bone marrow stromal cell spreading on trabecular bone structures. AB - The clinical application of macro-porous scaffolds for bone regeneration is significantly affected by the problem of insufficient cell colonization. Given the wide variety of different scaffold structures used for tissue engineering it is essential to derive relationships for cell colonization independent of scaffold architecture. To study cell population spreading on 3D structures decoupled from nutrient limitations, an in vitro culture system was developed consisting of thin slices of human trabecular bone seeded with Human Bone Marrow Stromal Cells, combined with dedicated microCT imaging and computational modeling of cell population spreading. Only the first phase of in vitro scaffold colonization was addressed, in which cells migrate and proliferate up to the stage when the surface of the bone is covered as a monolayer, a critical prerequisite for further tissue formation. The results confirm the model's ability to represent experimentally observed cell population spreading. The key advantage of the computational model was that by incorporating complex 3D structure, cell behavior can be characterized quantitatively in terms of intrinsic migration parameters, which could potentially be used for predictions on different macro-porous scaffolds subject to additional experimental validation. This type of modeling will prove useful in predicting cell colonization and improving strategies for skeletal tissue engineering. PMID- 19296222 TI - Analysis of musculoskeletal loadings in lower limbs during stilts walking in occupational activity. AB - Construction workers often use stilts to raise them to a higher level above ground to perform many tasks, such as taping and sanding on the ceiling or upper half of a wall. Some epidemiological studies indicated that the use of stilts may place workers at increased risk for knee injuries or may increase the likelihood of trips and falls. In the present study, we developed an inverse dynamic model of stilts walking to investigate the effects of this activity on the joint moments and musculoskeletal loadings in the lower limbs. The stilts-walk model was developed using the commercial musculoskeletal simulation software AnyBody (version 3.0, Anybody Technology, Aalborg, Denmark). Simulations were performed using data collected from tests of four subjects. All subjects walked without or with stilts through a 12-m straight path. The moments of the knee, hip, and ankle joints, as well as forces in major muscles or muscle groups in the lower limbs, for stilts walking were compared with those for normal walking. Our simulations showed that the use of stilts may potentially increase the peak joint moment in knee extension by approximately 20%; induce 15% reduction and slight reduction in the peak joint moments in ankle plantar flexion and hip extension, respectively. The model predictions on the muscle forces indicated that the use of stilts may potentially increase loadings in five of eight major muscle groups in the lower extremities. The most remarkable was the force in rectus femoris muscle, which was found to potentially increase by up to 1.79 times for the stilts walking compared to that for the normal walking. The proposed model would be useful for the engineers in their efforts to improve the stilts design to reduce musculoskeletal loadings and fall risk. PMID- 19296223 TI - Top manager effects on buprenorphine adoption in outpatient substance abuse treatment programs. AB - To examine the influence of top managers' characteristics on the adoption of buprenorphine for opioid dependence among U.S. outpatient substance abuse treatment units, this investigation analyzed a cross-sectional national study of 547 such units in the 2004-2005 wave of the Drug Abuse Treatment System Survey. Administrators reported their demographics, training, and treatment orientation, as well as features of the unit and its pattern of use of buprenorphine. Nationally, 15.8% of programs offered any buprenorphine services. Greater adoption of buprenorphine correlated with directors' younger age, longer tenure, male gender, and weaker endorsement of abstinence as the most important treatment goal. Availability of naltrexone and medical services also correlated positively with buprenorphine adoption. The authors conclude that leaders' characteristics are related to the adoption of innovative practices in addiction treatment programs. Future work should examine whether leadership development for community addiction programs might speed up the diffusion of buprenorphine and other innovative, evidence-based practices. PMID- 19296225 TI - Psychrometric analysis of the environmental equivalency factor for aqueous tablet coating. AB - Process control of aqueous tablet coating depends on a number of thermodynamic and psychrometric variables. Since many of these variables are interdependent, the choice of parameters by which to control the process or designate a design space is not necessarily obvious. Several mass or heat conservation models for aqueous tablet coating can be found in the literature, varying in approach and proposed method for controlling the coating process. A commonly used first principles model built upon the coupled heat and mass transfer in evaporative mass transfer derives an "Environmental Equivalency" (EE) factor as an indicator of the relative rate of water evaporation from the tablet bed surface and as a relevant scaling factor for aqueous coating. The EE factor is expressed by an equation involving ten individual parameters; however, if the derivation of EE is extended further under the context of an adiabatic process, a much-simplified yet equivalent expression for EE emerges consisting of only three parameters, each directly measurable or obtainable from a psychrometric chart and which bear direct significance to the gross thermodynamic conditions of the coating. The psychrometric model herein is presented as a more physically evocative description of the coating process, enhancing process understanding and potentially playing a key role in a Quality by Design approach to defining an aqueous coating design space. PMID- 19296224 TI - Development and in vivo floating behavior of verapamil HCl intragastric floating tablets. AB - A novel gastro retentive controlled release drug delivery system of verapamil HCl was formulated in an effort to increase the gastric retention time of the dosage form and to control drug release. Hydroxypropylmethylcellulose (HPMC), carbopol, and xanthan gum were incorporated for gel-forming properties. Buoyancy was achieved by adding an effervescent mixture of sodium bicarbonate and anhydrous citric acid. In vitro drug release studies were performed, and drug release kinetics was evaluated using the linear regression method. The optimized intragastric floating tablet composed of 3:2 of HPMC K4M to xanthan gum exhibited 95.39% drug release in 24 h in vitro, while the buoyancy lag time was 36.2 s, and the intragastric floating tablet remained buoyant for >24 h. Zero-order and non Fickian release transport was confirmed as the drug release mechanism from the optimized formulation (F7). X-ray studies showed that total buoyancy time was able to delay the gastric emptying of verapamil HCl intragastric floating tablet in mongrel dogs for more than 4 h. Optimized intragastric floating tablet showed no significant change in physical appearance, drug content, total buoyancy time, or in vitro dissolution pattern after storage at 40 degrees C/75% relative humidity for 3 months. PMID- 19296226 TI - Studies of the crystallization of amorphous trehalose using simultaneous gravimetric vapor sorption/near IR (GVS/NIR) and "modulated" GVS/NIR. AB - The purpose of this research was to investigate the influence of changes in the amorphous state on the crystallization of trehalose. Amorphous trehalose is known to stabilize biomaterials; hence, an understanding of crystallization is vital. Amorphous trehalose, prepared by spray-drying, was exposed to either a single step (0-75%) in relative humidity (RH) or to modulated 0-75-0% RH to cause crystallization. For the single-step experiment, two samples crystallized in a predictable manner to form the dihydrate. One sample, while notionally identical, did not crystallize in the same way and showed a mass loss throughout the time at 75% RH, with a final mass less than that expected for the dihydrate. The idiosyncratic sample was seen to have a starting near infrared (NIR) spectra similar to that exhibited by anhydrous crystalline trehalose, implying that short range order in the amorphous material (or a small amount of crystalline seed, not detectable using powder X-ray diffraction) caused the sample to fail to form the dihydrate fully when exposed to high RH. The modulated RH study showed that the amorphous material interacted strongly with water; the intensity of the NIR traces was not proportional to mass of water but rather the extent of hydrogen bonding. Subsequent crystallization of this sample clearly was a partial formation of the dihydrate, but with the bulk of the sample then shielded such that it was unable to show significant sorption when exposed to elevated RH. It has been shown that the nature of the amorphous form will alter the way in which samples crystallize. With oscillation in RH, it was possible to further understand the interactions between water and amorphous trehalose. PMID- 19296228 TI - Prediction of the deposition of dry powder aerosols. AB - The trajectory, mass fluxes, and deposition of aerosolized particles in a complex tubular system have been predicted. A procedure based on Lagrangian stochastic modeling is proposed to enable the anticipation of such phenomena, taking advantage of experimental results to characterize the air flow. The predictions have been obtained for pharmaceutical aerosols delivered by dry powder inhalers. A critical assessment of the dispersion model has been carried out using data available in the literature. The procedure assumes a low volume fraction of particles in the simulation of turbulent dispersion, but deposition is physically based on the interaction between the particles and both solid and liquid surfaces. The results were confirmed by experimental tests of powder deposition, run according to the European Pharmacopoeia. A parametric study was also carried out with the aim of providing a more complete evaluation of the model's performance. The comparison between predictions and experimental results has shown that the model properly describes the deposition of aerosolized particles. PMID- 19296227 TI - Targeted delivery systems for oligonucleotide therapeutics. AB - Oligonucleotides including antisense oligonucleotides and siRNA are emerging as promising therapeutic agents against a variety of diseases. Effective delivery of these molecules is critical to their successful clinical application. Targeted systems can greatly improve the efficiency and specificity of oligonucleotides delivery. Meanwhile, an effective delivery system must successfully overcome a multitude of biological barriers to enable the oligonucleotides to reach the site of action and access their biological targets. Several delivery strategies based on different platform technologies and different targeting ligands have been developed to achieve these objectives. This review aims at providing a summary and perspective on recent progress in this very active area of research. PMID- 19296229 TI - Molecular detection of Rickettsia, Coxiella and Rickettsiella DNA in three native Australian tick species. AB - Three Australian native animal species yielded 60 samples composed of three indigenous ticks. Hosts included twelve koalas, two echidnas and one wombat from Victoria, and ticks were of the species Ixodes tasmani (n = 42), Bothriocroton concolor (n = 8) and B. auruginans (n = 10), respectively. PCR screening and sequencing detected a species of Coxiella, sharing closest sequence identity to C. burnetii (>98%), in all B. auruginans, as well as a species of Rickettsia, matching closest to R. massiliae, in 70% of the same samples. A genotype sharing closest similarity to Rickettsia bellii (>99%) was identified in three female B. concolor collected from one of the echidnas. Three samples of I. tasmani, taken from three koalas, yielded different genotypes of Rickettsiella. These results represent the first detection of the three genera in each tick species and identify a high level of previously undetected bacterial diversity in Australian ticks. PMID- 19296230 TI - Monitoring deterioration of vegetation cover in the vicinity of smelting industry, using statistical methods and TM and ETM(+) imageries, Sarcheshmeh copper complex, Central Iran. AB - Simple statistical methods on Normalized Difference Vegetation Index (NDVI) and bands 3 and 4 data of relatively coarse resolution Thematic Mapper (TM) and Enhanced Thematic Mapper plus (ETM(+)) imageries were used to investigate the impacts of air pollution on the deterioration of the vegetation cover in the Sarcheshmeh copper complex of central Iran. Descriptive statistics and k-means cluster analysis indicated that vegetation deterioration had already started in the prevailing wind directions. The results show that combination of simple statistical methods and satellite imageries can be used as effective monitoring tools to indicate vegetation stress even in regions of sparse vegetation. Despite various possible perturbing factors upon NDVI, this index remains to be a valuable quantitative vegetation monitoring tool. PMID- 19296231 TI - CD90 Expression on human primary cells and elimination of contaminating fibroblasts from cell cultures. AB - Cluster Differentiation 90 (CD90) is a cell surface glycoprotein originally identified on mouse thymocytes. Although CD90 has been identified on a variety of stem cells and at varying levels in non-lymphoid tissues such as on fibroblasts, brain cells, and activated endothelial cells, the knowledge about the levels of CD90 expression on different cell types, including human primary cells, is limited. The goal of this study was to identify CD90 as a human primary cell biomarker and to develop an efficient and reliable method for eliminating unwanted or contaminating fibroblasts from human primary cell cultures suitable for research pursuant to cell based therapy technologies. PMID- 19296232 TI - Pharmacokinetics, urinary excretion and plasma protein binding of danofloxacin following intravenous administration in buffalo calves (Bubalus bubalis). AB - Pharmacokinetics, urinary excretion and plasma protein binding of danofloxacin was investigated in buffalo calves following intravenous administration at the dose rate of 1.25 mg/kg to select the optimal dosage regimen of danofloxacin. Drug concentrations in plasma and urine were measured by microbiological assaying. In vitro plasma protein binding was determined employing the equilibrium dialysis technique. The distribution and elimination of danofloxacin were rapid, as indicated by values (mean +/-SD) of distribution half-life (t(1/2)alpha = 0.16 +/- 0.07 h) and elimination half-life (t(1/2)beta = 4.24 +/- 1.78 h), respectively. Volume of distribution at steady state (Vss) = 3.98 +/- 1.69 L/kg indicated large distribution of drug. The area under plasma drug concentration versus time curve (AUC) was 1.79 +/- 0.28 micrg/ml x h and MRT was 8.64 +/- 0.61 h. Urinary excretion of danofloxacin was 23% within 48 h of its administration. Mean plasma protein binding was 36% at concentrations ranging from 0.0125 microg/ml to 1 microg/ml. On the basis of pharmacokinetic parameters obtained, it is concluded that the revision of danofloxacin dosage regimen in buffalo calves is needed because the current dosage schedule (1.25 mg/kg) is likely to promote resistance. PMID- 19296233 TI - Protein profile and alpha-lactalbumin concentration in the milk of standard and transgenic goats expressing recombinant human butyrylcholinesterase. AB - The expression of recombinant proteins of pharmaceutical interest in the milk of transgenic farm animals can result in phenotypes exhibiting compromised lactation performance, as a result of the extraordinary demand placed on the mammary gland. In this study, we investigated differences in the protein composition of milk from control and transgenic goats expressing recombinant human butyrylcholinesterase. In Experiment 1, the milk was characterized by gel electrophoresis and liquid chromatography/mass spectrometry in order to identify protein bands that were uniquely visible in the transgenic milk and/or at differing band densities compared with controls. Differences in protein content were additionally evaluated by computer assisted band densitometry. Proteins identified in the transgenic milk only included serum proteins (i.e. complement component 3b, ceruloplasmin), a cytoskeleton protein (i.e. actin) and a stress induced protein (94 kDA glucose-regulated protein). Proteins exhibiting evident differences in band density between the transgenic and control groups included immunoglobulins, serum albumin, beta-lactoglobulin and alpha-lactalbumin. These results were found to be indicative of compromised epithelial tight junctions, premature mammary cell death, and protein synthesis stress resulting from transgene expression. In Experiment 2, the concentration of alpha-lactalbumin was determined using the IDRing assay and was found to be significantly reduced on day 1 of lactation in transgenic goats (4.33 +/- 0.97 vs. 2.24 +/- 0.25 mg/ml, P < 0.01), but was not different from non-transgenic controls by day 30 (0.99 +/- 0.46 vs. 0.90 +/- 0.11 mg/ml, P > 0.05). We concluded that a decreased/delayed expression of the alpha-lactalbumin gene may be the cause for the delayed start of milk production observed in this herd of transgenic goats. PMID- 19296234 TI - Renal involvement in nail-patella syndrome: report of three cases. PMID- 19296235 TI - Direct renin inhibitors: ONTARGET for success? AB - Direct renin inhibitors are the first new class of antihypertensive to emerge since angiotensin II receptor blockers. We discuss their reno- and cardioprotective potential, based on extrapolation from animal models and phase three trials that are currently ongoing. This paper reviews the potential benefits of direct renin inhibitors (DRIs), the only new anti-hypertensive class developed in the last decade, as compared to pre-existing classes of drug inhibiting more downstream, such as Angiotensin Converting Enzyme inhibitors (ACEI), Angiotensin 2 Receptor Blockers (ARBS). PMID- 19296237 TI - Cloning and characterization of the Rubisco activase gene from Ipomoea batatas (L.) Lam. AB - A full-length cDNA of Rubisco activase (IBrcaI) was cloned from sweet potato (Ipomoea batatas (L.) Lam) using Rapid-Amplification of cDNA Ends (RACE). IBrcaI contains a 1,347 bp open reading frame encoding a protein of 439 amino acids. The sequence alignment of multiple Rubisco activase genes from sweet potato and other plants showed high homology at two previously described ATP-binding sites. Western blot analysis indicated that there are two Rubisco activase proteins in sweet potato. Expression of IBrcaI was only detected in leaves. In the 14 h light and 10 h dark photoperiods, maximal and minimal IBrcaI mRNA expression levels were detected at 8:00 in the morning and at midnight, respectively. PMID- 19296238 TI - Vitamin D receptor genotypes and kidney allograft rejection. AB - OBJECTIVE: Transplantation of renal grafts is an established treatment for renal failure in a variety of medical conditions. Polymorphisms in genes, coding for proteins involved in immune response, may influence immunological and non immunological mechanisms that lead to allograft loss. Vitamin D receptor (VDR) agonist has been shown to reduce short and long term allograft rejection in animal model. There are functional polymorphisms in VDR gene. MATERIALS AND METHODS: A total of 75 renal allograft recipients with at least 2 years follow-up were selected and genotyped for two polymorphisms in the VDR genes (FokI and BsmI) and the association of each genotype with renal allograft survival and acute rejection was evaluated. RESULTS: We are unable to find statistically significant association between any of the study polymorphisms and clinical outcomes. CONCLUSION: We have found no evidence to suggest that either VDR FokI or BsmI polymorphism determines the incidence of acute rejection or graft survival after renal transplantation. A larger sample size is necessary to confirm these findings. PMID- 19296239 TI - Intraoperative touch imprint cytology of sentinel lymph nodes in breast cancer: experience at a tertiary care center in Mexico. AB - BACKGROUND: Sentinel lymph node (SLN) biopsy in patients with breast cancer has emerged as a conservative and promising procedure. One of the most important issues is the intraoperative evaluation of the SLN with a high degree of accuracy by frozen section and/or imprint cytology. The objective of this study was to test the ability of intraoperative touch imprint cytology (ITIC) to predict metastasis on SLN. METHODS: SLNs were freshly examined, bisected in <0.5 cm or serially sectioned at 2 mm intervals on the long axis. Each surface of the section was touched on the glass slide, and stained. Results of ITIC were compared with permanent sections. Sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV), and accuracy (Acc) were calculated. False negatives were reviewed. RESULTS: We analyzed 179 SLN from 110 patients. The comparison between ITIC and final results of the SLN showed 139 (77.6%) true negative imprints, and 28 (15.6%) true positive. There were 12 (6.70%) false negative (FN) imprints which included 6 macrometastases, 3 micrometastases, and 3 isolated tumor cells. Re-screening after the definitive results of false negative imprints showed again 10 negative imprints, one with two groups of cells and one with multiple groups of cells. The overall Se was 70% (73.6% for micro/macrometastases and 82.3% for macrometastases), Sp and PPV were 100% in all cases. NPV was 92.1% overall (93.4% micro/macrometastases and 96% in macrometastases).Global accuracy was 93.3% (94.4% for micro/macrometastases and 96% for in macrometastases). CONCLUSIONS: ITIC is excellent to detect macrometastases, however, it fails to detect micrometastases. False negative imprints for macrometastases are mainly due to sampling error. The immediate availability, low cost, high Sp, PPV, preservation of the lymph node for histopathologic examination, avoiding of a second surgery are the major advantages of intraoperative evaluation of SLN. PMID- 19296240 TI - 18F-FDG-PET imaging of a benign neurofibroma masquerading as metastatic oesophageal carcinoma. AB - We present the case of a 65-year-old male with squamous cell carcinoma of the thoracic oesophagus on a background of neurofibromatosis type 1. On computed tomography, he was noted to have a large left-sided superior mediastinal mass. Initially, this mass was thought to be metastatic lymphadenopathy; however, it did not display fluorodeoxyglucose uptake on positron emission tomography. Subsequent biopsy confirmed the mass to be a neurofibroma and the patient commenced definitive chemo-radiation. Positron emission tomography had a major impact on management since the presumed lymph node disease was not included in the radiation field. In addition, positron emission tomography altered prognostic stratification since lymph node involvement is a poor prognostic factor in oesophageal cancer. We could only identify one other case in the English literature in which positron emission tomography was used to distinguish metastatic carcinoma from a neurofibroma, although there are a number of reports that describe the utility of positron emission tomography in differentiating benign neurofibromas from malignant connective tissue tumours. PMID- 19296241 TI - [Clinical analysis of superficial spreading early gastric cancer:a report of 62 cases]. AB - OBJECTIVE: To clarify the clinicopathological features and prognosis, and to evaluate the rational surgical therapy for superficial spreading early gastric cancer (EGC). METHODS: Superficial spreading EGC was defined as tumor invading the mucosa or submucosa layer with the diameter> or =50 mm, and common EGC as tumor with diameter < 50 mm. The clinicopathological data of 62 patients with superficial spreading EGC and 224 patients with common EGC were collected and analyzed retrospectively. RESULTS: No significant differences in age, sex, differentiation, depth of invasion, growth manner, vessel involvement and lymph node metastasis were found between superficial spreading and common EGC(P>0.05), but significant differences were seen in tumor site, macroscopic type, scope of gastrectomy and lymphadenectomy between the two groups(P<0.05). Superficial spreading EGC located in the middle or the whole of stomach accounted for 45.2%, and presented mixed macroscopic type more frequently(48.4%), such as II(C+II(A 5 cases, II(C+II(B 9 cases and II(C+III( 16 cases, and more total gastrectomy and more D(2) lymphadenectomy(16.1% and 40.3% respectively) were executed, compared with common EGC. Due to cancer residual,two patients underwent enlargement of the scope of gastrectomy,two underwent total gastrectomy and one underwent operation again after surgery. The 5-, 10-year tumor-free survival rates for superficial spreading EGC were 98.4% and 91.4%, while 97.0% and 91.2% for the common EGC (chi(2)=1.16,P=0.282). CONCLUSIONS: Special clinicopathological features can be seen in superficial spreading EGC, and the lesion is superficial and extensive. Prevention of cancer residual in remnant stomach is the key point and D(2) lymphadenectomy is suitable enough for operation. PMID- 19296242 TI - [Living anatomical observations on peripancreatic spaces and their implications on laparoscopic gastrectomy with D(2) lymphadenectomy for distal gastric cancer]. AB - OBJECTIVE: To explore living anatomy of pancreas and peripancreatic spaces,as well as their implications on laparoscopic gastrectomy with D(2) lymphadenectomy for distal gastric cancer. METHODS: Living observation was carried out in 132 patients diagnosed as distal gastric cancer and undergoing laparoscopic gastrectomy with D(2) lymphadenectomy. RESULTS: Spaces between greater omentum and transverse mesocolon continued to pre-pancreatic and retro-pancreatic spaces at inferior margin of pancreas. The pre-pancreatic and retro-pancreatic spaces continued each other at inferior and superior margin of pancreas and extended in all directions. Left gastroepiploic vessels were located in pre-pancreatic spaces at superior margin of pancreatic tail. In retro-pancreatic space at inferior margin of pancreatic neck, superior mesenteric veins were located. In retro pancreatic spaces or in gastric mesenteries inferior to gastric antrum, right gastroepiploic vessels were located. In spaces between gastric antrum and pancreatic heads, gastroduodenal arteries were located and traced to locate common hepatic arteries. In retro-pancreatic spaces at superior margin of pancreatic body, common hepatic arteries, left gastric arteries,celiac arteries and splenic arteries were located. Hepatopancreatic folds and gastropancreatic folds were landmarks respective to locate common hepatic arteries and left gastric arteries. The aforementioned vessels and spaces in their vagina vasorum continued each other and united as a whole. CONCLUSIONS: Laparoscopic gastrectomy with D(2) lymphadenectomy for distal gastric cancer is carried out in macroscopic surgical planes of pre-pancreatic space and retro-pancreatic space, as well as their extensions in all directions, and in microscopic surgical planes of spaces in vagina vasorum of perigastric vessels which continue each other, under the guidance of central landmarks of pancreas and concrete landmarks of vessel trunks and their furcations. PMID- 19296243 TI - [Efficacy of combined splenectomy in proximal gastric cancer with No.10 lymph node metastasis]. AB - OBJECTIVE: To explore the impact on prognosis of D(2) lymphadenectomy combined with splenectomy in patients of advanced proximal gastric cancer with No.10 lymph node metastasis. METHODS: Clinical data of 216 patients of advanced proximal gastric cancer with No.10 lymph node metastasis undergone D(2) curative resection in our hospital from January 1980 to December 2002 were analyzed retrospectively. Among them, 73 underwent simultaneous splenectomy (splenectomy group), while 143 without splenectomy (spleen-preserving group). The 5-year survival rate, the mean numbers of dissected No.10 lymph nodes and metastatic No.10 lymph nodes, the complication morbidity and mortality were compared between the two groups. RESULTS: The 5-year survival rates of splenectomy group and spleen-preserving group were 30.0% and 19.7% respectively, whose difference was significant(P<0.05). The mean numbers of dissected No.10 lymph nodes and metastatic No.10 lymph nodes in splenectomy group were significantly greater than those in spleen-preserving group(P<0.05). Splenectomy, invasion depth and gastrectomy type were independent prognostic factors. The survival rates of T(3) patients in splenectomy group and spleen-preserving group were 38.7% and 18.9% respectively, whose difference was significant (P<0.05). The survival rates of patients undergone total gastrectomy in splenectomy group and spleen-preserving group were 33.4% and 20.7% respectively, whose difference was significant (P<0.05). The complication morbidity and mortality in splenectomy group were 24.7% and 4.1%, while in spleen-preserving group were 17.5% and 3.5% respectively, whose differences were not significant(P>0.05). CONCLUSIONS: Splenectomy is benefit for No.10 lymph node dissection in patients with advanced proximal gastric cancer. To improve the prognosis, total gastrectomy combined with splenectomy should be recommended for patients of T(3) proximal gastric cancer with No.10 lymph node metastasis. Simultaneous splenectomy dose not increase the complication morbidity and mortality. PMID- 19296244 TI - [Application of neoadjuvant chemotherapy in laparoscopic gastrectomy for advanced gastric cancer]. AB - OBJECTIVE: To explore the benefit of neoadjuvant chemotherapy in advanced gastric cancer patients treated by laparoscopy. METHODS: Fifteen patients with histologically proved gastric adenocarcinomas (stages II(, III(, IIII(M(0)) were treated with FOLFOX7 neoadjuvant chemotherapy followed by laparoscopy between June 2005 and March 2007( trial group). Thirty patients were assigned to the control group with only laparoscopic treatment in the same period. The clinicopathological data were compared between two groups. RESULTS: All the patients in trial group accepted four cycles of preoperative chemotherapy and the toxicity was less than grade 3. Two of them achieved complete response, 10 achieved partial response and 3 kept stable disease. Ten patients of trial group underwent laparoscopic-assisted radical gastrectomy. The rates of R(0) resection(80.0%) and pN(0) (60.0%) in trial group were significantly higher than those in control group(46.7% and 20.0%), while the rate of positive lymph node 11.0%(34/309) was significantly lower than that of control group 23.8%(142/596). The operation time and postoperative complication were similar in two groups. CONCLUSIONS: Advanced gastric cancer after neoadjuvant chemotherapy can be down regulated in the stage, increase the rate of R(0)-resection, diminish the infiltration extent of tumor, decrease the metastasis of lymph node, and increase the possibility of laparoscopic radical gastrectomy. PMID- 19296245 TI - [Multivariate analysis of risk factors of lymph node metastasis in early gastric cancer]. AB - OBJECTIVE: To investigate the clinicopathological factors affecting lymph node metastasis in early gastric cancer(EGC). METHODS: Clinicopathological data of 369 patients with EGC from Jan. 1999 to Jun. 2008 were analyzed retrospectively. Multivariate analysis was performed to find the risk factors affecting lymph node metastasis. RESULTS: The age, gender, tumor size, invasion depth, histological classification and vascular cancer thrombosis were associated with lymph node metastasis. Multivariate analysis showed that tumor size and depth of invasion were main independent risk factors. CONCLUSIONS: The main independent risk factors affecting lymph node metastasis in EGC are tumor size and invasion depth. The gender, vascular cancer thrombosis, tumor size, invasion depth and histological classification should be considered to establish the surgical program. PMID- 19296246 TI - [Evaluation the value of number of metastatic perigastric lymph nodes on prognosis in patients with gastric cancer]. AB - OBJECTIVE: To find out a simple and reproductive prognostic index in gastric cancer patients,which can be used as a comparable parameter among different regions of China. METHODS: The perigastric metastatic lymph nodes(PGMLN) with long-term survival data were retrospectively evaluated in 148 gastric cancer patients, undergone potentially curative resections. RESULTS: The 3-year cumulative survival rate was 62.8% and the survival rate decreased significantly with the increase of PGMLN, especially when the PGMLN was more than 6. The PGMLN had a clear linear relationship with total positive lymph nodes(r=0.94, P<0.01), while it had little correlation with total resected lymph nodes(r=0.18,P=0.2). CONCLUSIONS: PGMLN is a convenient and reliable prognostic factor in gastric cancer patients, which may be a good candidate of comparable parameters among different regions of China. PMID- 19296247 TI - [Risk factors of intra-abdominal infection following gastrectomy in gastric cancer patients]. AB - OBJECTIVE: To investigate the main risk factors associated with intra-abdominal infection(IAI) following gastrectomy in gastric cancer patients. METHODS: Case control study was used to investigate the clinical data of 1728 gastric carcinoma cases retrospectively by Logistic regressive analysis. RESULTS: Univariate Logistic regressive analysis showed 16 factors, including age, malnutrition, chronic obstructive pulmonary disease(COPD), diabetes mellitus(DM), heart diseases, prothrombin time, lymphocyte count, tumor size, ascites, invasion to the adjacent organ, neoplasm TNM staging (UICC, 1997), methods of gastrectomy, blood loss, operative time, blood transfusion and extent of lymph nodal dissection,were associated with postoperative intra-abdominal infection. Binary Logistic regression analysis found that extent of lymph nodal dissection(N(2)(+) approximately N(3) and N(2)), invasion to the adjacent organ, DM, operative time, age and lymphocyte count were the independent risk factors associated with mortality. CONCLUSION: Necessary interventions should be carried out to prevent IAI referring to above risk factors. PMID- 19296248 TI - [Roles of nutrition risk screening and preventive enteral nutritional support before radical resection of gastric cancer]. AB - OBJECTIVE: To investigate the potential role of the preventive nutritional support in patients with nutritional risk defined by nutrition risk screening 2002(NRS 2002) before radical resection of gastric cancer. METHODS: Patients with gastric cancer were evaluated by NRS 2002 preoperatively. Elective patients with nutritional risk were randomly assigned into 3 d preventive enteral nutrition(EN) group versus control group. The preventive nutrition regimen was 1000 ml Nutrison Multi Fibre(4184 kJ/L). The changes in body weight lost, serum albumin, immunoglobulin were recorded postoperatively. RESULTS: One week after operation, the preventive EN group showed less decrease in body weight as compared to control group, which was statistically significant. The levels of serum albumin and IgA on day 1 and day 3 after operation in preventive EN group were significantly higher than those in control group. CONCLUSION: Before operation for gastric cancer, patients with nutritional risk defined by NRS 2002 may benefit from preventive enteral nutrition, which improves the patients' nutritional condition and enhances their immunologic function. PMID- 19296249 TI - [Expression of transcription factor SP1, vascular endothelial growth factor and CD34 in serosa-infiltrating gastric cancer and their relationship with biological behavior and prognosis]. AB - OBJECTIVE: To investigate the expression of transcription factor SP1, vascular endothelial growth factor(VEGF) and CD34 in serosa-infiltrating gastric cancer and their relationship with biological behavior and survival rate. METHODS: Immunohistochemical technique was used to detect the expression of SP1, VEGF and CD34(described by microvessel density, MVD) in 68 specimens with serosa infiltrating gastric cancer. RESULTS: The positive expression rates of SP1 and VEGF in serosa-infiltrating gastric cancer were 50.0% and 52.9% respectively. In positive SP1 specimens, the positive rate of VEGF(73.5%) was significantly higher than that of negative SP1 specimens (32.4%, chi(2)=11.57, P=0.01). The mean tumor MVD was correlated with the expression levels of SP1 and VEGF(P<0.01). There was a significant correlation of the SP1 expression with tumor size and growth pattern(P =0.01). The expression levels of VEGF and MVD were correlated with Borrmann types, cell differentiation, metastatic lymph nodes and growth pattern(P<0.01). Univariate analysis revealed that SP1 and VEGF expression, MVD, Borrmann types, lymph node metastasis, tumor size and growth pattern were significant prognostic factors related to survival time. Multivariate analysis showed that SP1 expression, MVD and growth pattern were independently prognostic factors of poor survival. CONCLUSIONS: The activation of SP1 contributes to angiogenesis and metastasis in gastric cancer through the up-regulation of VEGF. SP1, VEGF and MVD may serve as valuable indicators of biological behavior of gastric cancer. SP1 protein expression is not related with the number of metastatic lymph nodes. SP1 expression and MVD may serve as valuable indicators of prognosis in gastric carcinoma. PMID- 19296250 TI - [Analysis of clinicopathology and prognosis in 181 patients with gastrointestinal stromal tumors]. AB - OBJECTIVE: To investigate the therapeutic experience of gastrointestinal stromal tumors (GIST) and to analyze the pathological features and prognostic factors of GIST. METHODS: The clinicopathological and follow-up data of 181 patients with GIST admitted in Renji Hospital between January 1999 and December 2007 were analyzed retrospectively. All the cases were grouped according to Fletcher's risk scheme. Life table and COX regression model were used to evaluate the prognostic factors. RESULTS: Out of 181 tumors, 107(59.1%) were located in stomach, 51 (28.2%) in intestine and 23(12.7%) in colorectum or other sites. Distant metastases,including liver metastases were found in 7 patients intraoperatively. Tumor size ranged from 0.5 to 30 cm with the mean of 7.02 cm. The positive rate of CD117 was 94.5% (171/181) and that of CD34 was 86.2% (156/181). One hundred and seventy-six patients underwent complete resections, including multi-organ resections in 26 patients. The other patients underwent palliative operations. The 1-, 3- and 5-year overall survival rates of 181 patients were 95.2%, 87.9% and 78.5% respectively. Univariate analysis revealed age, tumor size, primary organ of tumor, mitotic count, Fletcher's classification and multi-organ resection were associated with survival rate. No significant difference of sex was existed among groups. COX hazard proportional model revealed that advanced stage and large tumor size indicated worse prognosis. Eight patients with high risk of recurrence and 3 patients with recurrence and metastasis were stable after receiving imatinib therapy. CONCLUSIONS: The diagnosis of GIST depends on endoscope and CT. Fletcher's classification is simple and effective to evaluate GIST behavior and prognosis. Surgical resection is still the main therapy for GIST and targeted therapy will play a more important role for prognosis in the future. PMID- 19296251 TI - [Surgical treatment for patients with advanced gastrointestinal stromal tumor after targeted therapy]. AB - OBJECTIVE: To explore the role of surgery and its long-term outcome in patients with advanced gastrointestinal stromal tumor(GIST) treated with imatinib preoperatively. METHODS: Thirteen patients receiving imatinib therapy preoperatively, were retrospectively assessed for completeness of surgical resection and for disease-free and overall survival after resection. RESULTS: Thirteen patients, including 3 patients with locally advanced primary GIST and 10 patients with recurrent or metastatic GIST, underwent surgery after preoperative treatment with imatinib. Complete resections were accomplished in 4 of the 5 responsive disease(RD) patients, and in 1 of the 8 progression disease(PD) patients (38.5%). The progression-free survival(PFS) time for patients with RD and PD were 24.8 months and 2.8 months respectively. The difference of PFS between patients with RD and those with PD was significant(P<0.01). Median overall survival(OS) was not reached in both patients with RD and PD. The difference of OS between patients with RD and those with PD was not significant(P>0.05). CONCLUSION: Surgical intervention following imatinib is feasible and can be considered for patients with advanced GIST responsive to imatinib. PMID- 19296252 TI - [A clinical study on the degree of mesorectal tumor invasion in middle-low rectal cancer]. AB - OBJECTIVE: To study the relationship between the degree of mesorectal tumor invasion and prognosis of the patients with middle-low rectal cancer. METHODS: Specimens from 49 patients with middle-low rectal cancer, undergone total mesorectal excision in our hospital from April 2003 to December 2003,were studied by large slice pathologic technique. The thickness of mesorectum and the depth of tumor infiltration were measured, and the degree of mesorectal tumor invasion was calculated. The local recurrence rate, metastasis rate and 5-year survival rate were investigated respectively. Possible clinicopathological influence factors were also analyzed. RESULTS: The local recurrence rate was 12.2%(6/49), the distant metastasis rate was 26.5%(13/49). In three different degrees of mesorectal tumor invasion(I(,II(,III(), the local recurrence rates were 0, 7.7% and 31.3% (chi(2)=7.357, P =0.015); the metastasis rates were 10%, 23.1% and 50%(chi(2)=7.405, P =0.025); the 5-year survival rates were 90.9%, 69.2% and 28.6%(log-rank, P =0.013). Tumor diameter, T and N staging were risk factors influencing the degree of mesorectal tumor invasion(chi(2)=6.849 P=0.033, chi(2)=34.845 P =0.000, chi(2)=17.266 P =0.002). CONCLUSIONS: The degree of mesorectal tumor invasion is an important predictor of local and distant metastasis as well as survival of patients in middle-low rectal carcinoma. The degree of mesorectal tumor invasion in the middle-low rectal carcinoma is significantly correlated with tumor diameter, T and N stage. PMID- 19296253 TI - [Evaluation of capsule endoscopy in the diagnosis of small bowel disease]. AB - OBJECTIVE: To evaluate the clinical utility of capsule endoscopy in diagnosing small bowel disease. METHODS: A retrospective review of 155 cases undergone capsule endoscopy examinations from September 2002 to March 2007 in our hospital were performed. Gastric and small bowel transit time, patient tolerability, number of complete and incomplete examinations, and examination findings were evaluated. RESULTS: A total of 155 cases were reviewed, and 159 times of capsule endoscopy examination were finished. Of the 155 patients, 97 suffered from obscure gastrointestinal bleeding, 42 abdominal pain, 6 abdominal discomfort, 4 diarrhea, and 6 body check. The capsule endoscopy examination for the entire small bowel were finished in 93.1%(148/159)cases. All the patients had no discomfortable feeling during the examinations. The capsule endoscopy remained in the stomach for an average of 65.5 min(1 to 335 min). The mean transit time in the small bowel was 282.2 min(45 to 524 min). The diagnostic yield of capsule endoscopy was 78.6%(125/159). Vasculopathy was present in 43.4% patients, enteritis in 28.3%, submucous knot in 10.1%, diverticulum of small intestine in 8.2%, and small intestine tumor in 5.7%. Other findings consisted of polyposis of small intestine, foreign objects and parasite. The diagnostic yield of obscure gastrointestinal bleeding by capsule endoscopy was 89.7%, and of abdominal pain was 73.8%. CONCLUSIONS: Capsule endoscopy is safe and well tolerated. Capsule endoscopy is a valuable diagnostic tool in the evaluation of occult small bowel disease, especially for obscure gastrointestinal bleeding. PMID- 19296254 TI - [Efficacy of polyglycosides of Tripterygium wilfordii in preventing postoperative recurrence of Crohn disease]. AB - OBJECTIVE: To observe the efficacy of polyglycoside of Tripterygium wilfordii(GTW) in preventing postoperative recurrence of Crohn disease(CD). METHODS: Thirty-nine post-operative CD patients in whom all of the diseased gut had been removed from January 2005 to December 2006 were enrolled in a randomized, placebo-controlled trial. The patients took GTW(21 cases) or SASP(18 cases) in two weeks after operation. Crohn disease activity index(CDAI), ESR and CRP were collected at week 0, 13, 26, 52 or at the onset of symptoms. Ileocolonoscopy was performed at the end of the trial or at the onset of symptoms. RESULTS: One patient in GTW group and 2 patients in SASP group were lost and 2 patients in GTW were excluded from the trial for non-compliance. Clinical recurrence was ascertained in one patient (5.6%) received GTW and in four (25.0%) received SASP. Four of eighteen patients in GTW(22.2%) had endoscopic recurrence compared with nine of sixteen(56.2%) in SASP. There were significant differences between the two groups(P<0.05). CONCLUSION: Tripterygium wilfordii showed good efficacy in preventing recurrence of postoperative CD which can maintain remission and prevent recurrence. PMID- 19296255 TI - [Clinical analysis of rectal neuroendocrine carcinoma:a report of 16 cases]. AB - OBJECTIVE: To study the clinical data and surgical treatment strategy of rectal neuroendocrine carcinoma (NEC). METHODS: Sixteen cases of rectal NEC and 222 cases of rectal carcinomas receiving surgical treatment in Beijing Cancer Hospital from 2003 to 2007, were analyzed retrospectively. RESULTS: Among the 16 rectal NEC patients, 4(25%) received Miles surgery, 7(43.8%) Dixon surgery, 2 combined multiple organ resection and 3 local resection. Lymph note metastases occurred in 11 cases(68.8%) and distant metastases in 7 cases (43.8%). Among the 222 rectal carcinoma patients, 43(19.4%) received Miles surgery, 152(68.5%) Dixon surgery, 12 palliative operation, 6 colostomy and 9 just received laparotomy. Lymph note metastases occurred in 125 cases (56.3%). In rectal NEC group, postoperative 1-, 2- and 3-year survival rates were 62.5%, 25.0% and 0.63% respectively, which were significantly lower than 83.1%, 61.7% and 46.1% in rectal carcinoma group(all P<0.01). CONCLUSIONS: Rectal NEC is a rare disease. More vascular invasion, lymph node and distant organ metastases are found in rectal NEC than rectal carcinoma, and the prognosis of rectal NEC is worse than rectal cancer. PMID- 19296256 TI - [Evaluation of the primary lesion detection in colorectal carcinoma with (18)F FDG PET-CT]. AB - OBJECTIVE: To explore the clinical value of detecting primary lesions of colorectal carcinoma with (18)F-FDG PET-CT. METHODS: Clinical data of 23 cases of colorectal carcinoma collected from April 2007 to June 2008, including PET-CT, endoscopy, operation and pathology, were analyzed retrospectively. RESULTS: Of the 23 colorectal cancer patients, including 15 males and 8 females, 11 received abdominal CT contrast examination, 7 abdominal CT general examination and 5 chest CT examination. The lesions located in caecum in 5 cases, ascending colon 4 cases, transverse colon 2 cases, descending colon 2 cases, sigmoid colon 4 cases, rectum 6 cases. CT images showed local mass, incrassation and nodes in colon. PET images revealed intensely hypermetabolic lesions. The maximum of standard uptake value (SUVmax) was 11.7+/-9.5, and the delay SUVmax was 14.8+/-11.0. The bigger was the mass, the higher was the SUV. The highest SUV was 44.8. Metastatic lesions were found in 15 cases. More cases and more metastatic lesions were found by (18)F-FDG PET-CT. CONCLUSION: The primary and metastatic lesions of colorectal carcinoma can be detected sensitively and exactly by (18)F-FDG PET-CT, which is helpful in tumor staging and making the treatment plan. PMID- 19296257 TI - [Bowel preparation with sodium phosphate versus polyethylene glycol: a prospective, randomized, controlled clinical trial]. AB - OBJECTIVE: To compare the efficacy, tolerance and safety between oral sodium phosphate(NaP) and polyethylene glycol(PEG) on bowel preparation. METHODS: One hundred and fifteen inpatients were randomly divided into NaP group and PEG group. The questionnaire was designed for scoring by patients and doctors regarding to tolerance, taste, side effects and cleaning degree etc. RESULTS: Compared with PEG group, NaP presented better tolerance, lower side effects and higher rate of adequate cleaning quality(P<0.05). NaP could cause electrolytic alterations, such as hyperphosphatemia, hypernatremia, hypocalcemia and hypopotassemia, but these changes were transient and without clinical significance. CONCLUSION: Sodium phosphate is safe and effective for bowel preparation, and is better than polyethylene glycol in tolerance. PMID- 19296258 TI - [Efficacy of cellulose on functional constipation]. AB - OBJECTIVE: To assess the clinical efficacy and safety of cellulose on functional constipation. METHODS: A prospective, self-controlled, multicenter clinical trial of cellulose was conducted for 2 weeks in 240 patients with functional constipation according to the Rome III( criteria. Symptoms and characters of feces before and after the treatment were observed and evaluated according to a score scheme. RESULTS: In the 240 patients, the frequencies of defecation increased and the characters of feces was improved significantly after 2-week treatment. There were no adverse reactions observed throughout the clinical trial. The total efficacy was 82.1% at day 7 and 90.7% at day 14. The satisfactory rate of doctors was 83.8% and of patients was 83.8%. CONCLUSION: Cellulose is effective and safe in the treatment of chronic functional constipation. PMID- 19296259 TI - [Clinicopathological significance of expression and amplification of P21 activated kinase 1 gene in colorectal carcinoma]. AB - OBJECTIVE: To investigate the clinicopathological value of the expression and amplification of P21-activated kinase 1 gene (PAK1) in colorectal carcinoma(CRC). METHODS: Immunohistochemistry (IHC), fluorescence in situ hybridization (FISH) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL) methods were used to examine the protein expression, amplification of PAK1 and cell apoptosis in 80 cases of CRC and 30 cases of colorectal adenoma by tissue microarray. RESULTS: IHC showed an overexpression of PAK1 protein in 26% of colorectal adenomas and 62% of CRCs. Significant association was found between expression of PAK1 and tumor histological grade as well as tumor clinical stage(P<0.05). In poor-differentiated(G(3)) CRCs, PAK1 expression in 90% carcinoma was up-regulated, which was significantly higher than that in tumors of G(1/2)(51%). Overexpression of PAK1 was detected in 78% of CRCs in later clinical stages (Dukes C, D), which was significantly higher than that in early clinical stages (Dukes A,B, 53%). In addition, negative correlation between PAK1 overexpression and cell apoptosis was observed in these CRC cohorts(P<0.05). FISH revealed that amplification of PAK1 gene was examined in only 3% CRCs. CONCLUSIONS: Overexpression of PAK1 protein may play an important role in development and progression of colorectal neoplasms and it is closely associated with the malignant histological and invasive phenotype of CRCs. The expression of PAK1 in CRC may be used as one of the new molecular markers in predicting tumors malignant potential and progression. PMID- 19296260 TI - [Tissue-engineering nerve repairs pelvic autonomic nerve defects in Beagle dogs]. AB - OBJECTIVE: To investigate the outcome of repairing pelvic autonomic nerve defects with the tissue-engineered nerve, in order to provide a new method and experimental evidence for solving sexual disturbance induced by pelvic autonomic nerve injury after radical resection of rectal carcinoma. METHODS: Bone marrow stromal cells (BMSCs) were purified with density gradient centrifugation. A 10 mm defect of hypogastric nerve was created in 9 Beagle dogs and the 18 hypogastric nerves were randomly divided into three groups. Group A: nerve defects bridged with copolymer of lactic and glycolic acids (PLGA) tube containing BMSCs and collagen protein sponge. Group B: with PLGA tube only containing collagen protein sponge. Group C: with autologous nerve graft. The effect of nerve recovery was evaluated by morphology, HE staining, neurofilament immunohistochemistry staining, electron microscope scanning and measurement quantity of new axon 12 weeks after the transplantation. RESULTS: Twelve weeks after the transplantation, degradation of PLGA tubes showed in group A and group B. The nerves regenerated through defect area to distal end. The density of regeneration nerve fiber in group A and group C were better than that in group B. The difference was significant between group A or group C and group B (P<0.05), and no significant difference was observed between group A and group C(P>0.05). CONCLUSION: Tissue engineered nerve, which is constructed by BMSCs mixed with collagen protein sponge and PLGA tube, can be used to bridge and repair the pelvic autonomic nerve defect. PMID- 19296261 TI - [Effect of balsalazide on intestinal mucosal permeability of dextran sulfate sodium-induced colitis in mice]. AB - OBJECTIVE: To investigate the effect of balsalazide on intestinal mucosal permeability of experimental colitis induced by dextran sulfate sodium(DSS) in a mouse model and its possible mechanism. METHODS: Forty-five C57BL/6J mice were randomly divided into five groups. Normal group was only fed with distilled water, DSS group and Balsalazide groups at doses of 42,141,423 mg/kg were fed with 5% DSS. Balsalazide was given by intragastric administration. DAI was evaluated daily. At the end of the experiment, colon tissue was collected for assessment of histological changes, MDA content, MPO, SOD and GSH-PX activity. Small intestinal mucosa was collected for assessment of transmission electron microscope(TEM), and detection of permeability by Evans blue. RESULTS: Compared with normal group, DSS group mice all manifested severe weight loss associated with hematochezia and diarrhea with significant increase of DAI and HI score(P<0.01). MDA content and MPO activity was increased with the reverse result of SOD and GSH-PX(P<0.01) in DSS group. Intestinal mucosa showed a focal reduction in thinning of microvillous carpet and even a total disarrangement of epithelial surface, with decurtated and broaden junctional complex and enlarged intercellular space under TEM observations in DSS group. The amount of Evans blue permeated into intestinal wall was obvious in DSS group. Compared with DSS group, balsalazide improved gross findings, decreased MPO activity and MDA content, but increased the activity of SOD and GSH-PX(P<0.05). The amount of Evans blue permeated into intestinal wall was less(P<0.05). Ileal microvillous carpet was ameliorated in dose-dependent manner by balsalazide. CONCLUSIONS: Intestinal mucosal permeability is increased in DSS group. Balsalazide can significantly ameliorate intestinal mucosal permeability in colitis model. PMID- 19296262 TI - Evolution of the nutritional situation of indigenous and non-indigenous Chilean schoolchildren. AB - BACKGROUND: Latin American countries show accelerated but ethnically or socially differentiated changes in their epidemiological profiles. AIM: The present study examined the evolution of the nutritional situation (1997-2005) in Chilean schoolchildren as related to ethnical origin (Mapuche). SUBJECTS AND METHODS: Using official databases, stunting (height/age or=95 percentile) were ascribed in first-grade schoolchildren. Ethnic groups were assigned by native parents' surnames (none, one and two). RESULTS: Based on 1 757 155 children (average age: 76.3 months), in 1997 stunting reached 8.4%, 4.8% and 3.1% in children with two, one and no Mapuche surnames, respectively. In 2005 it fell to 3.7%, 3.1% and 2.6% - a marked decrease in those with two Mapuche surnames (p<0.001). Obesity in 1997 was 11.8%, 12.8% and 13.3%, whilst in 2005 it changed to 17.5, 18.5 and 18.6%, respectively, demonstrating a similar trend to obesity in all groups (p=0.153). Undernutrition was rare (<1.1%) and stable. Poverty decreased clearly among the Mapuche population in this period. CONCLUSION: The marked decrease in stunting in children with a strong indigenous background seems related to a decrease in poverty over the period. Yet, the increase of obesity in all groups deserves further analysis. PMID- 19296263 TI - Hurricane Katrina's impact on the mental health of adolescent female offenders. AB - Exposure to multiple traumatic events and high rates of mental health problems are common among juvenile offenders. This study draws on Conservation of Resources (COR) stress theory to examine the impact of a specific trauma, Hurricane Katrina, relative to other adverse life events, on the mental health of female adolescent offenders in Mississippi. Teenage girls (N=258, 69% African American) were recruited from four juvenile detention centers and the state training school. Participants were interviewed about the occurrence and timing of adverse life events and hurricane-related experiences and completed a self administered mental health assessment. Hierarchical linear regression models were used to identify predictors of anxiety and depression. Pre-hurricane family stressors, pre-hurricane traumatic events, hurricane-related property damage, and receipt of hurricane-related financial assistance significantly predicted symptoms of anxiety and depression. Findings support COR theory. Family stressors had the greatest influence on symptoms of anxiety and depression, highlighting the need for family based services that address the multiple, inter-related problems and challenges in the lives of female juvenile offenders. PMID- 19296264 TI - Predicting posttraumatic growth among Hurricane Katrina survivors living with HIV: the role of self-efficacy, social support, and PTSD symptoms. AB - The study tested the model of adaptation after trauma by Benight and Bandura (2004) indicating that posttraumatic recovery may be predicted directly by coping self-efficacy (CSE) and indirectly by social support. These relations were investigated in the context of posttraumatic growth (PTG) among Hurricane Katrina survivors living with HIV. Additionally, it was hypothesized that among individuals with more intensive Posttraumatic Stress Disorder (PTSD) symptoms, those with strong CSE would experience the strongest PTG. Cross-sectional data were collected among 90 patients with HIV who reinitiated care at the HIV outpatient clinic. Questionnaires were administered approximately 14 months after the hurricane. Higher CSE was related to higher PTG among the survivors who suffered from more intensive PTSD symptoms. Received social support was directly related to only one index of PTG, relating to others. Furthermore, although there was a significant relationship between social support and CSE, the indirect conditional effect of received social support on PTG was not confirmed. Similar results were obtained across the indices of PTG, controlling for the level of exposure to hurricane-related trauma. Cross-sectional design and convenience character of the sample warrants replications. PMID- 19296265 TI - Delivering youth-specific mental health services: the advantages of a collaborative, multi-disciplinary system. AB - OBJECTIVE: Evidence suggests that quality mental health care is based on well integrated multi-disciplinary care provided by a range of mental health, substance use, and general healthcare clinicians. There is a growing focus in Australia on providing this type of mental health care to young people, particularly those in the early stages of a major disorder. The development of such services has proceeded on the basis of limited service-based data and has also been impeded by current healthcare funding structures. METHODS: This report outlines the service characteristics of three models: a traditional 'fee for service' model, a specialized youth mental health clinic, and a new headspace multi-disciplinary site in South Western Sydney. RESULTS: Naturalistic data from these three services collected during their developmental phase indicate that each model is associated with differential demographic, illness and service organization characteristics. CONCLUSIONS: Compared with 'fee-for-service' type care, specialized youth models provide greater access to a broad range of multi disciplinary clinicians. PMID- 19296266 TI - Accountability or responsibility? The challenge of policies in clinical psychiatry. AB - OBJECTIVE: The aim of this paper is to describe some of the differences between personal and administrative approaches to clinical risk management by reflecting on some recent experiences in policy development. CONCLUSIONS: Administrative preoccupation with policy development and political accountability may detract from professional development and personal responsibility. PMID- 19296267 TI - RANZCP experiences in establishing and progressing an externally focussed mental health policy and partnership agenda. AB - OBJECTIVE: This paper traces the background of involvement and increasing external activity of the Royal Australian and New Zealand College of Psychiatrists (RANZCP) in the policy and project area. CONCLUSIONS: Throughout 2008, representatives of the RANZCP paused to review progress and celebrate successes in the implementation of the College's stated aim to develop an externally focussed policy agenda. A summary of key activities across the past 4 5 years is outlined, including the key leadership role undertaken by the RANZCP in the recent formation of the Mental Health Professionals Association's partnership. Tracing the history of the establishment of the RANZCP externally focussed policy functions, the paper provides details of the extensive range of projects and activities undertaken to date. Also highlighted is the broad range of resources that have been developed and are now available for College members including trainees. The progress achieved to date is reflective of RANZCP's deliberate strategic actions and intent to work more closely with others and take an increasingly active role in the development of mental health policy and service delivery standards across the Australian and New Zealand landscape. The functions are now embedded with the College's newly revised Strategic Plan. PMID- 19296268 TI - A Prison Mental Health Screening Unit: a first for New South Wales. AB - OBJECTIVE: The aims of this paper are to provide a description of a newly available service the Mental Health Screening Unit (MHSU), within the NSW prison system and to present the first 12 months in terms of the patients, its adaptations and its limitations. METHODS: The first section of this paper provides a thorough description of the MHSU. The second looks at data collected for the period 1 July 2006 to 30 June 2007. A variety of databases were utilized as sources. RESULTS: There were a total of 604 admissions. The most common primary diagnoses were schizophrenia-related disorders. There were relatively few adverse incidents, and 18% of the acute unit population were transferred to the Long Bay Prison Hospital. CONCLUSION: The MHSU had a considerable throughput of patients and managed a range of severe mental illnesses. The initial objectives of the MHSU have broadly been achieved. There are several future areas of research discussed. PMID- 19296269 TI - The PTSD supremacy: Criterion F in three Voyager cases. AB - OBJECTIVE: The aim is to consider whether the courts and experts in their application of Criterion F for posttraumatic stress disorder (PTSD) have applied it consistently in civil claims brought years after the event. METHOD: Three cases for compensation relating to the Voyager disaster are considered. RESULTS: It appears, from the cases considered in this paper, that while courts consider that Criterion F is crucial in making a diagnosis of PTSD, there are some inconsistencies in terms of understanding and applying this criterion, both by the courts and experts. This may be because of a lack of guidance in psychiatric texts as to how to apply Criterion F. CONCLUSIONS: Criterion F is, after the satisfaction of Criterion A(1), arguably the most important of the criteria for PTSD for, while the symptoms referred to in Criteria B-D have been shown to be fairly easy to simulate, it is arguably harder to do this with Criterion F, particularly in cases that arise a long time after the event. It is important therefore that psychiatrists assessing persons so long after an event adhere rigorously to Criterion F, because it is based on facts open to objective corroboration, while criteria B, C and D tend to rely on subjective experiences which are the most sensitive to distortion. Thus, it is to be hoped that if DSM-V is to maintain a criterion of clinically significant distress or impairment in the majority of the disorders described therein, it will provide some assistance as to how this criteria should be applied. PMID- 19296270 TI - Gender-sensitive mental health care. AB - OBJECTIVE: The aim of this paper is to examine aspects of mental health and mental health care through a gender lens. CONCLUSION: Gender differences have an impact on mental health and the experience and course of women's mental illness. Comprehensive gender-sensitive mental health care requires the planning, delivery, monitoring and quality improvement initiatives of mental health care to be informed by a knowledge and understanding of gender differences in women and men and their inter-relationship with respect to childhood and adult life experiences (e.g. violence and abuse); day-to-day social, cultural, and family realities; expression and experience of mental ill health and treatment needs and responses. PMID- 19296271 TI - Predicament suicide: concept and evidence. AB - OBJECTIVE: The aim of this paper is to introduce the term 'predicament suicide' and add to the evidence that suicide may occur in the absence of Axis I diagnoses. METHOD: The term 'predicament suicide' was described as suicide which occurs when the individual without mental disorder is in unacceptable circumstances from which they cannot find an acceptable alternative means of escape. Accounts of suicide that satisfied these criteria, and could be illustrated by images, were sought. RESULTS: Four accounts were identified: Ajax, Brutus, Mayor Freyberg and Commander Donnicke. CONCLUSIONS: Predicament suicide appears to be a useful designation. This work added evidence indicating that suicide may occur in the absence of an Axis I mental disorder. PMID- 19296272 TI - Theory of mind and psychiatry: an introduction. AB - OBJECTIVE: 'Theory of mind' (ToM) arose from the study of primates and their social organization, and scholars in many fields - philosophy, anthropology, psychology, psychiatry and neuroscience - have contributed to this expanding topic. In this paper, we provide an overview of aspects of ToM of relevance to psychiatry. We briefly describe the origins of ToM in primates and humans and some relevant neurobiology, and then touch on possible contributions to psychopathology. METHOD: We searched for articles on PubMed and Medline, using the terms 'theory of mind', 'mirror neuron system' and 'psychiatry'. CONCLUSION: There is evidence that ToM deficits are important in certain psychiatric disorders. While more research is required, an appreciation of ToM will have an impact on our further understanding and management of at least some mental disorders, including autism and schizophrenia. PMID- 19296273 TI - The Resource Team: an innovative service delivery support model for mental health services. AB - OBJECTIVE: This paper outlines the development of the Resource Team, an innovative service delivery model supporting clinical services at the Inner North Brisbane Mental Health Service, Royal Brisbane and Women's Hospital Health Service District. The team aims to provide a base for specialist mental health support staff, improve knowledge management and support the development of meaningful community partnerships. METHOD: Development of the team included a literature review and consultation with internal and external stakeholders. From this, the objectives, roles and functions of the team were clarified and disseminated to stakeholders. RESULTS: The team currently encompasses 12 positions and has initiated a number of programs and service developments. These include improved IT management of clinical resources and the development of partnerships with the community and non-government sectors. CONCLUSIONS: The Resource Team effectively coordinates specialist clinical support positions, addresses knowledge management issues and facilitates meaningful engagement with the community and non-government sectors. The model could easily be applied in other mental health and general health services. PMID- 19296274 TI - Liability of mental health services for injuries incurred during community treatment. AB - OBJECTIVES: The aim of this paper is to examine the recent decision of the NSW Supreme Court, which considered a personal injuries action brought by a patient treated in the community following a 6 day voluntary hospital admission. CONCLUSIONS: The judicial reasoning in Walker v Sydney West Area Health Service provides some comfort to mental health professionals practising in other jurisdictions whose legislative provisions are similar to those contained in the Civil Liability Act (NSW). In applying the Bolam principle, rather than the higher common law standard previously imposed by the High Court in Rogers v Whitaker, the decision is encouraging for mental health professionals whose management accords with accepted current good practice. The infrequent scrutiny by courts and coroners of management practice and systems in mental health is a further incentive to maintain continuous improvement of quality of care (by clinical audits, active risk management, professional development and supervision, and patient/family participation) consistent with the principles of clinical governance. PMID- 19296275 TI - Service user and significant other versions of the Health of the Nation Outcome Scales. AB - OBJECTIVES: The aim of this paper was to test the measurement properties of self report (SR) and significant other (SO) versions of the Health of the Nation Outcome Scales (HoNOS). METHOD: SR and SO HoNOSs, with content closely paralleling the clinician HoNOS but with item 8 expanded into scales rating each difficulty separately, were developed and trialed with Early Intervention for Psychosis service users and their significant others. RESULTS: Satisfactory agreement was found between SR, SO and clinician HoNOSs, and between the SR and SO on most additional items. CONCLUSIONS: Results suggest adequate validity of the measures. The additional items proved useful for identifying comorbidities that may have been under-recognized. Results suggest that SR and SO versions of the HoNOS can be valid and valuable for triangulating and recording the perspectives of different stakeholders in promoting recovery. PMID- 19296276 TI - ECT: shocked beyond belief. AB - OBJECTIVE: The aim of this paper is to consider electroconvulsive therapy (ECT) in the 21st century and whether it should be a treatment of last resort from the perspective of someone who has received the treatment in recent years. CONCLUSION: It seems that informed consent, the introduction of advance directives and the improved delivery of ECT have done little to ease people's fear, with a lot of people still believing ECT to be an extreme form of treatment and only a last resort option. The idea of any treatment being considered 'last resort' in mental health care gives the impression that it is the most radical and worst treatment available - thus we leave it until last to try. Yet for many people, including myself, the treatment is a valid and preferred option with minimal side effects. PMID- 19296277 TI - A response to Plastow. PMID- 19296278 TI - From the president. PMID- 19296279 TI - From the honorary secretary. PMID- 19296280 TI - From the chief executive officer. PMID- 19296281 TI - Obituary. PMID- 19296283 TI - Criminal law as it pertains to 'mentally incompetent defendants': a McNaughton rule in the light of cognitive neuroscience. AB - The McNaughton rules for determining whether a person can be successfully defended on the grounds of mental incompetence were determined by a committee of the House of Lords in 1843. They arose as a consequence of the trial of Daniel McNaughton for the killing of Prime Minister Sir Robert Peel's secretary. In retrospect it is clear that McNaughton suffered from schizophrenia. The successful defence of McNaughton on the grounds of mental incompetence by his advocate Sir Alexander Cockburn involved a profound shift in the criteria for such a defence, and was largely based on the then recently published 'scientific' thesis of the great US psychiatrist Isaac Ray, entitled 'A treatise on the medical jurisprudence of insanity'. Subsequent discussion of this defence in the House of Lords led to the McNaughton rules, still the basis of the defence of mental incompetence in the courts of much of the English-speaking world. This essay argues that the rules need to be reconsidered in the light of the discoveries of cognitive neuroscience made during the 160 years since Ray's treatise. It is shown, for instance, how the conflation of 'the power of self control' with 'irresistible impulse' by Cockburn is not supported by cognitive neuroscience because these are separate capacities requiring normal activity in distinct brain structures for their expression. In this way cognitive neuroscience assists in distinguishing between different capacities. It is further shown that failure of appropriate restraint in the expression of a capacity can be related to failure of synapses in particular parts of the brain. This raises the question as to what level of synaptic loss will the legislature and the courts rule as sufficient for a subject to be no longer held responsible for their lack of restraint. PMID- 19296284 TI - Randomized controlled trial of an early intervention programme for adolescent anxiety disorders. AB - OBJECTIVE: To assess the effectiveness of an indicated early intervention and prevention programme for anxiety disorders when conducted by school staff. METHOD: A total of 260 students in their first year of high school with self reported anxiety symptoms >1 SD above the mean score of a normative sample were randomly allocated on the basis of their school to an intervention condition led by school staff or to a monitoring condition. RESULTS: There was little difference between conditions at the 2 year and 4 year follow up on self-reported symptoms, and no difference on diagnosis or health-care use. CONCLUSIONS: Outcomes associated with indicated prevention programmes led by school staff may not be as strong as those produced by specialist mental health staff. More work is needed to delineate models by which evidence-based treatments can be effectively used in schools. PMID- 19296285 TI - Internet-based course for the management of stress for junior high schools. AB - OBJECTIVE: To determine the feasibility and efficacy of a Web-based stress management programme for schools. METHOD: A six-lesson Web-based programme was constructed in collaboration with the schools. It was implemented across Year 8 in six schools (464 students). Students completed measures of knowledge, perceived competence, and coping behaviours, prior to the programme; at the conclusion of the last lesson; and 3 months later. Well-being and distress were measured prior to each lesson. RESULTS: Input from schools into the development of the programme was very valuable. Knowledge about stress improved after the six lessons and students reported that support seeking coping had increased and avoidance behaviour decreased. Scores on measures of distress went down and well being scores rose. Seventy-nine per cent of the students completed five or more lessons. CONCLUSIONS: Web-based curriculum-consistent education in mental health is feasible and efficacious at the level expected from a brief universal intervention. PMID- 19296286 TI - Anterior hippocampus in schizophrenia pathogenesis: molecular evidence from a proteome study. AB - OBJECTIVE: The purpose of the present study was to identify differentially expressed proteins in the anterior and posterior hippocampus of brains of schizophrenia patients compared to neurologically healthy controls. METHOD: Proteins extracted from fresh frozen post-mortem posterior and anterior hippocampus for nine schizophrenia and nine control individuals, and seven schizophrenia and seven control individuals, respectively, were screened for differential expression using 2-D gel electrophoresis and mass spectrometry. RESULTS: A significantly larger number of protein spots were differentially expressed in the anterior (n = 43) compared to the posterior (n = 16) hippocampus, representing 34 and 14 unique proteins, respectively. These proteins are involved in cytoskeleton structure and function, neurotransmission and mitochondrial function. CONCLUSION: Based on the aberrant protein expression profiles, the anterior hippocampus appears to be more involved in schizophrenia pathogenesis than the posterior hippocampus. Furthermore, consistent with previous findings, we found molecular evidence to support abnormal neuronal cytoarchitecture and function, neurotransmission and mitochondrial function in the schizophrenia hippocampus. PMID- 19296288 TI - Validation of an Australian sign language instrument of outcome measurement for adults in mental health settings. AB - OBJECTIVE: There are currently no adult mental health outcome measures that have been translated into Australian sign language (Auslan). Without a valid and reliable Auslan outcome measure, empirical research into the efficacy of mental health interventions for sign language users is unattainable. To address this research problem the Outcome Rating Scale (ORS), a measure of general functioning, was translated into Auslan and recorded on to digital video disk for use in clinical settings. The purpose of the present study was therefore to examine the reliability, validity and acceptability of an Auslan version of the ORS (ORS-Auslan). METHOD: The ORS-Auslan was administered to 44 deaf people who use Auslan as their first language and who identify as members of a deaf community (termed 'Deaf' people) on their first presentation to a mental health or counselling facility and to 55 Deaf people in the general community. The community sample also completed an Auslan version of the Depression Anxiety Stress Scale-21 (DASS-21). RESULTS: t-Tests indicated significant differences between the mean scores for the clinical and community sample. Internal consistency was acceptable given the low number of items in the ORS-Auslan. Construct validity was established by significant correlations between total scores on the DASS-21-Auslan and ORS-Auslan. Acceptability of ORS-Auslan was evident in the completion rate of 93% compared with 63% for DASS-21-Auslan. CONCLUSIONS: This is the only Auslan outcome measure available that can be used across a wide variety of mental health and clinical settings. The ORS-Auslan provides mental health clinicians with a reliable and valid, brief measure of general functioning that can significantly distinguish between clinical and non clinical presentations for members of the Deaf community. PMID- 19296287 TI - Short-term outcomes of mothers and newborn infants with comorbid psychiatric disorders and drug dependency. AB - OBJECTIVES: The aim of the present study was to determine the characteristics and short-term outcomes of mother-infant pairs with comorbid drug dependency and psychiatric disorders. METHODS: A population-based retrospective chart review was carried out of 879 drug-dependent mother and infant pairs in New South Wales (NSW) and the Australian Capital Territory (ACT) who delivered between 1 January and 31 December 2004. RESULTS: Psychiatric comorbidity (dual diagnosis, DD) was identified in 396 (45%) of the 879 drug-dependent women. DSM-IV depression (79%), followed by anxiety (20%), was most prevalent. DD women were more likely to use amphetamines (29% vs 18%, p < 0.05), less likely to use opiates (42% vs 51%, p < 0.05) and to have had no antenatal care (24% vs 8%, p < 0.05). They also had more previous pregnancies (4, range = 2-5 vs 3, range = 2-5, p < 0.05) and domestic violence (29% vs 14%, p < 0.05) was more common. DD infants were less likely to be admitted to a nursery (47% vs 55%, p < 0.05). Withdrawal scores were similar (maximum median Finnegan scores = 4 (interquartile range = 3-8) vs 10 (interquartile range = 7-12, p = 0.30) but fewer needed withdrawal medication (19% vs 27%, p < 0.05). Maternal psychotropic agents did not worsen the severity of neonatal withdrawal. CONCLUSIONS: Psychiatric comorbidity, especially depression, is common and affects almost half of drug-using mothers. Antenatal care, drug use and social outcomes are worse for DD mothers and their infants. It is recommended that all drug-using women be assessed antenatally for psychosocial disorders so that timely mental health intervention can be offered, if required. PMID- 19296289 TI - Changes in use of psychostimulant medication for ADHD in South Australia (1990 2006). AB - OBJECTIVE: The aim of the present study was to undertake a retrospective analysis of archival data on psychostimulant prescriptions from South Australia for the periods 1990-2000 and 2001-2006 for 7849 youths aged from birth to 18 years. METHOD: A person-based data set was used to assess: (i) rate of new prescriptions by age group; (ii) demographic characteristics (age of psychostimulant start, male: female ratio); (iii) duration of psychostimulant use; and (iv) geographic variation in psychostimulant prescription. RESULTS: Four major findings were observed: (i) the rate of new prescriptions was highly variable both for 1990 2000 and 2000-2006; (ii) demographic characteristics such as start age and male:female ratio declined over both periods; (iii) the duration of psychostimulant use was approximately 2.5 years for 1990-2000 and 2.0 years for 2000-2006; and (iv) there was geographic variation in both periods with a significant correlation between socioeconomic status and prescription rate per region. CONCLUSIONS: The patterns of psychostimulant use in Australia closely parallel the USA. Physicians' prescribing practice may be extremely volatile. Duration of psychostimulant treatment should receive increased attention. There is pronounced geographic variability in prescription rates, which may be related to socioeconomic status. PMID- 19296290 TI - Confirmatory factor analysis of the Parental Bonding Instrument in a Brazilian female population. AB - OBJECTIVE: The Parental Bonding Instrument (PBI) is a widely used measure of parenting. Recent studies have proposed different factor structures. There is a disagreement in the literature about whether the PBI is best used as a two-factor or a three-factor measure. METHOD: Two hundred and fifty-seven female adults were recruited from a clinical population (139 psychiatric patients and 118 controls) and were requested to complete the PBI. Maximum likelihood confirmatory factor analyses were performed to compare the five different factor structures in terms of model fit. RESULTS: The poorest fit to the data was obtained by the Parker et al. model. The Kendler model was the only model that had an adjusted goodness-of fit index >0.8 regarding both paternal and maternal PBI. When considering invariance of factor structure across age subgroups, the Kendler model was also the only acceptable model. CONCLUSION: Three-factor structures are preferable to two-factor structures. The Kendler model was the only one to provide an acceptable fit, but it must be considered that it was a female sample, and when considering gender subgroups other studies have found the same results. Despite the gender limitation, the present study contributes to a better understanding and use of the PBI in Brazilian samples. PMID- 19296291 TI - Management of depression by general practitioners: impact of physician gender. AB - OBJECTIVE: The aim of the present study was to determine whether anecdotal claims of gender differences in the treatment of depression by general practitioners (GPs) existed in practice. METHOD: Referral letters from 100 GPs to a specialized psychiatric depression clinic were analysed by word count and gender of referrer. Second, a Web-based survey of 517 participants examined the impact of GP gender in terms of levels of management nuances. RESULTS: The first study established that female GPs wrote distinctly longer referral letters. The second study identified that female GPs were seen as distinctly more caring over a range of parameters and identified the impact of some GP-patient gender differences. CONCLUSIONS: Reasons why female GPs are viewed as more caring - and any impact on the management of those with a depressive disorder - would benefit from refined investigation. PMID- 19296292 TI - Changing utilization of pro re nata ('as needed') sedation in a child and adolescent psychiatric inpatient unit. AB - OBJECTIVE: The primary purpose of the present study was to examine changes in utilization of pro re nata (PRN; 'as required') sedation over time within a child and adolescent psychiatric inpatient unit. The secondary purpose was to assess whether changes in PRN sedation were related to changing patient characteristics. METHODS: A retrospective chart review examined 257 medical charts from a child and youth mental health inpatient service over two time periods (wave I, n = 122; wave II, n = 135) and collected data on PRN sedation, patient characteristics and routine medications. RESULTS: Over time a significant reduction was observed in the proportion of patients prescribed PRN sedation from 70% to 54% (p < 0.01), and a reduction in the proportion of patients given PRN sedation from 46% to 26% (p < 0.01). The most commonly administered drug was chlorpromazine in wave I, and diazepam in wave II. Multivariate analysis indicated that reductions in PRN sedation occurred independently of changes in patient characteristics. CONCLUSIONS: High utilization rates of PRN sedation are not inevitable in a child and adolescent psychiatric inpatient unit and may be reduced over time. Changing utilization of PRN sedation occurred independently from changing patient characteristics. More treatment outcome studies are required to optimize use of PRN sedation in young people. PMID- 19296293 TI - Post-traumatic stress disorder and health problems among medically ill Canadian peacekeeping veterans. AB - OBJECTIVE: The aim of the present study was to examine the relationship between post-traumatic stress disorder (PTSD) symptom severity and four significant health conditions (gastrointestinal disorders, musculoskeletal problems, headaches, and cardiovascular problems). METHOD: Participants included 707 Canadian peacekeeping veterans with service-related disabilities, from a random, national Canadian survey, who had been deployed overseas. RESULTS: PTSD severity was significantly related to gastrointestinal disorders, musculoskeletal problems, and headaches, but not to cardiovascular problems. Controlling for demographic factors did not affect PTSD's relationships with the three significant health conditions. CONCLUSIONS: The present study supports previous work in finding consistent relations between PTSD severity and specific types of medical problems. PMID- 19296294 TI - Vicarious trauma, secondary traumatic stress or simply burnout? Effect of trauma therapy on mental health professionals. AB - OBJECTIVES: The aim of the present study was to perform an assessment for secondary traumatic stress (STS), vicarious trauma (VT) and workplace burnout for Australian mental health professionals involved in clinical practice. METHODS: Recruited directly by mail, randomly selected participants were invited to submit a questionnaire by post or online. Of the 480 participants contacted, 152 mental health professionals completed the questionnaire, which contained measures of STS, VT and burnout. RESULTS: Exposure to patients' traumatic material did not affect STS, VT or burnout, contradicting the theory of the originators of STS and VT. Rather, it was found that work-related stressors best predicted therapist distress. CONCLUSIONS: These findings have significant implications for the direction of research and theory development in traumatic stress studies, calling into question the existence of secondary trauma-related phenomena and enterprises aimed at treating the consultants. PMID- 19296295 TI - Effect of augmentatory repetitive transcranial magnetic stimulation on auditory hallucinations in schizophrenia: randomized controlled study. AB - OBJECTIVE: Auditory hallucinations are a characteristic symptom of schizophrenia and are usually resistant to treatment. The present study was conducted to further support the findings that repetitive transcranial magnetic stimulation (rTMS) reduces auditory hallucinations, and to evaluate the effect of low frequency rTMS on auditory hallucinations in schizophrenia. METHODS: Forty schizophrenia patients were included in the study. Patients were randomized to control or experimental group. Low-frequency rTMS (1 Hz, 90% motor threshold) was applied to the left temporoparietal cortex of patients in the experimental group for 10 days following the standard guidelines as an addition to antipsychotic treatment. The control group received only antipsychotics. The changes in the psychopathology scores for the auditory hallucinations were recorded using auditory hallucination recording scale. The rater was blind to the intervention procedure. RESULTS: A significant improvement was found in auditory hallucinations in the experimental group as compared to the control group. CONCLUSION: Left temporoparietal rTMS warrants further study as an intervention for auditory hallucinations. Data suggest that this intervention selectively alters neurobiological factors determining frequency of these hallucinations. PMID- 19296296 TI - Consciousness and hallucinations in schizophrenia: secondary aspects of generalized neuropil pathology? PMID- 19296297 TI - The expression and prognostic significance of Cks1 in salivary cancer. AB - Cks1 is an essential factor in facilitating Skp2-dependent degradation of p27, but its role in salivary malignancies is unknown. Expression of cyclin-dependent kinase subunit 1 (Cks1) was examined in 64 salivary malignancies, compared with p27, S-phase kinase protein 2 (Skp2), Ki-67, p53, and TDT-mediated dutp-biotin nick end labeling (TUNEL) expression, and with THE patient's clinical and pathological parameters. Cks1 expression was markedly increased in 30 patients (47%) and strongly correlated with increased expression of Skp2, Ki-67, p53, and TUNEL, but inversely with p27 levels. High expression of Cks1 WAS strongly associated with lymph node metastases and poor prognosis and survival. Cks1 alterations may have a significant biological role in the pathogenesis of salivary cancer. PMID- 19296298 TI - Methylphenidate, interstimulus interval, and reaction time performance of children with attention deficit/hyperactivity disorder: a pilot study. AB - Thirteen children with attention deficit/hyperactivity disorder (ADHD: DSM-IV-TR) participated in the pilot study. They carried out a Go/No-Go test with a short (2 seconds) and long (6 seconds) interstimulus interval (ISI) when on placebo and a therapeutic dose of methylphenidate (MPH). For the long-ISI placebo condition the responses were slow and inaccurate. This pattern of response may be due to underactivation of the readiness-to-respond state that is not fully controlled by effort allocation. Speed of response and accuracy were enhanced during the short ISI placebo condition and the long-ISI MPH condition. However, the combined effect (short ISI and MPH) resulted in a fast but inaccurate response style. This pattern of response may be due to overactivation of the readiness-to-respond state. The data of the pilot study support the stimulus shift hypothesis: MPH administration result in deterioration on tests on which children had previously done well (short ISI plus placebo versus short ISI plus MPH). In addition, the data support the idea that ADHD is associated with poor state regulation rather than motivational (delay aversion) theories or temporal-processing/time estimation theories of ADHD. The pilot study defined empirically an issue for further study with the larger controlled sample. PMID- 19296299 TI - Righting errors in writing errors: the Wing and Baddeley (1980) spelling error corpus revisited. AB - We present a new analysis of our previously published corpus of handwriting errors (slips) using the proportional allocation algorithm of Machtynger and Shallice (2009). As previously, the proportion of slips is greater in the middle of the word than at the ends, however, in contrast to before, the proportion is greater at the end than at the beginning of the word. The findings are consistent with the hypothesis of memory effects in a graphemic output buffer. PMID- 19296301 TI - Mechanical stretch regulates the expression of matrix metalloproteinase in rheumatoid arthritis fibroblast-like synoviocytes. AB - Mechanical stretch plays a crucial role in articular joints. In rheumatoid arthritis (RA), it is well known that fibroblast-like synoviocytes (FLS) produce matrix metalloproteinases (MMPs), resulting in local invasion into and degradation of bone and cartilage. We sought to examine whether mechanical stretch regulates the expression and underlying signal pathways of MMP secretion (MMP-1, -3, -13) in RA-FLS. FLS were grown on elastic silicone membrane in an equibiaxial strain apparatus and were exposed to 6% mechanical stretch (equivalent to gentle stretch exercise) for 15 min and 75 min, respectively. Semiquantitative PCR and real-time PCR were used to measure and analyze gene expression. Protein levels were determined by Western blotting. The results showed that 15 min of mechanical stretch inhibited MMP-1 and MMP-13 mRNA and protein level. However, the degree of inhibition by 75 min of stretch in expression of MMP-1 and MMP-13 was lower compared with 15 min stretch groups. The mRNA expression of ERK-1, ets-1 and citied-2 were increased by 6% mechanical stretch under both time points, however c-jun and c-fos mRNA level were affected differently after 15 min and 75 min mechanical stretch compared to control group. There were no significant changes on MMP-3 and ets-2 mRNA level under both 6% mechanical stretch time points. In the presence of pro-inflammatory cytokines (IL 1beta and TNF-alpha), the stretch also reduced the mRNA expression of MMP-1 and MMP-13. In short, our results showed that gentle mechanical strain affects MMP-1 and MMP-13 expression, potentially through the ERK-1-ets-1-cited-2-c-jun signaling pathway. PMID- 19296302 TI - Role of collagen fibers in acupuncture analgesia therapy on rats. AB - Acupuncture, a traditional Chinese therapeutic technique, has been put into practice for more than 4000 years and widely used for pain management since 1958. However, what is the mechanism underlying the acupuncture for analgesia effects by stimulation of acupoints, what substances receive the original mechanical acupuncture signals from the acupoints, or what transforms these signals into effective biological signals are not well understood. In this work, the role of collagen fibers at acupoints during acupuncture analgesia on rats was investigated. When the structure of the collagen fibers at Zusanli (ST36) was destroyed by injection of type I collagenase, the needle force caused by the acupuncture declined and the analgesic effects of rotation or lift-thrusting manipulations was attenuated accompanying the restraint of the degranulation ratios of mast cells. We propose that collagen fibers play an important role in acupuncture-induced analgesia, and they participate in signal transmission and transform processes. PMID- 19296303 TI - In vivo effects of ovarian steroid hormones on the expressions of estrogen receptors and the composition of extracellular matrix in the anterior cruciate ligament in rats. AB - Female athletes have a significantly higher rate of anterior cruciate ligament (ACL) injury than their male counterparts. Sex steroid hormones are considered to have an influence as risk factors for female ACL injuries. We hypothesized that estrogen and progesterone have specific and synergistic influences on the composition of extracellular matrix in ACL. By ovariectomy (OVX) followed by subcutaneous estradiol (E2) and/or progesterone (P4) replacement, 40 female rats were divided into 5 groups: E2, P4, combined E2 and P4 (EP), OVX control, and sham group. After 30 days, using undecalcified sections of knee joints in conjunction with immunofluorescence staining of estrogen receptor alpha and beta (ERalpha and ERbeta), collagen types 1 and 3, and cartilage oligomeric matrix protein (COMP), the immunoreactivities of these proteins in two distinct parts of ACL, proximal and middle portions, were compared semiquantitatively among experimental groups. By E2 replacement, the expressions of ERalpha in ACL fibroblasts were elevated compared to the OVX group. At the proximal portion, the immunoreactivities of type 1 collagen by E2 replacement, type 3 collagen by P4 replacement, and COMP by E2 or P4 replacement were significantly reduced. At the middle portion, the immunoreactivity of type 3 collagen was significantly elevated by E2 replacement. However, no differences were observed between the sham and OVX groups. These findings suggest that ACL is ER-dependent and that ovarian hormones alter ligament tissue composition, especially at the proximal portion. Female hormonal influences are partly involved in the biological properties of ACL. PMID- 19296304 TI - Role of matrix metalloproteinases in skin ageing. AB - Matrix metalloproteinases are a family of ubiquitous endopeptidases playing a role in many different physiological and pathological processes in the skin. They are also involved in cutaneous ageing. This review summarizes the features and regulation of these enzymes and presents an overview of the molecular mechanisms of both intrinsic and extrinsic skin ageing presents. The role of matrix metalloproteinases in skin ageing is discussed in detail. PMID- 19296305 TI - This special issue of Culture, Health and Sexuality, contains papers from the IASSCS last conference in Lima in June 2007. Introduction and welcome. PMID- 19296306 TI - Intersexuality and sexual rights in southern Brazil. AB - This paper reports of the socio-medical management of intersexuality in a teaching hospital in southern Brazil. Findings reveal that the 'search for sex' in the body treats sex as a medical-diagnostic category constructed during decision-making negotiations. In the course of the various bodily 'regulations' and 'corrections' imposed, the insufficiency of a model in which categories are based around a sexual dichotomy is revealed. By questioning this dichotomy, it is possible to examine theoretical and ethical limits circumscribing the field of bioethics and the notion of sexual rights as human rights. The paper also explores the tensions that can arise from the application of sexual rights to health issues in general, and the medical sphere in particular. PMID- 19296307 TI - Have they really come out: gay men and their parents in Taiwan. AB - In Chinese culture, filial piety for a son is closely linked to his capacity to produce an heir to ensure continuity of the paternal line. For Taiwanese gay men, coming out as gay may be interpreted as a refusal to produce a male heir and thus constitutes a major conflict within their family. This study explores how gay men in Taiwan come out to their parents within this cultural context. Thirty-two men in total were interviewed. Findings demonstrate that the decision to come out was often motivated by the son's perception of his parents' attitude towards homosexuality. Respondents worked hard to prepare for coming out and to minimize the risk and the impacts of the process, their report shows that some parents go through their own process of coming out and/or hiding in the closet after their gay son's coming out. Although many parents still see homosexuality as illness, some adopt alternative discourses to reinterpret the meaning of being gay as a spiritual path to eternal enlightenment or friendship. These findings imply sites of resistance to the privileged discourse of filial piety in constituting the experiences of coming out for Taiwanese gay men and their parents. PMID- 19296308 TI - Black lesbian gender and sexual culture: celebration and resistance. AB - Lesbian gender expression is a persistent theme in research and writing about lesbian culture. Yet little empirical research has examined the ways lesbian gender functions within the sexual culture of lesbian communities, particularly among lesbians of colour. This study was aimed at documenting and assessing the functions of lesbian gender among African American lesbians. Particular attention was paid to identifying core characteristics of sexual discourses, such as evidence of dominant and resistant sexual scripts and contradictions between messages about sex. This study took the form of a rapid ethnography of an African American lesbian community in the USA using focus groups, individual community leader interviews and participant observations at a weekly open mic event. Findings document how lesbian gender roles translated into distinct sexual roles and expectations that appear to both parallel and radically reject heterosexual norms for sex. The deep roots of the social pressure to date within these roles were also evident within observations at the open microphone events. While data highlighted the central role that lesbian gender roles play in this community, analyses also revealed a strong resistance to the dominance of this sexual cultural system. PMID- 19296309 TI - Liminal identities: Caribbean men who have sex with men in London, UK. AB - Accounts by 10 Caribbean men who have sex with men living in the UK reveal them to be liminal beings with unstable and unresolved identities. They are between social states: aware they are not heterosexual and not publicly recognised, or in some cases self-accepted, as homosexual. Caribbean-born respondents especially suffer from homophobia, expressing regret and disappointment at their sexuality. They may also experience cognitive dissonance - as they are aware of their conflict with the heteronormative order - they cannot resolve. Religion contributes to homophobia and cognitive dissonance particularly for Caribbean born men, some of whom may believe a fundamental conflict exists between Christianity and homosexuality. Heterosexism and homophobia contribute to and reinforce their liminal state, by preventing transition to publicly recognised homosexual status. Respondents may engage in private and public, internal and external, overt and covert policing of their and other gay men's behaviour: through strategic pretence at heterosexuality and/or condemnation of men engaging in behaviour identifiable as stereotypically homosexual, for example. Narratives point to the need to complexify the conventional understanding of Jamaican heterosexism to explain reported variations in the degree of anti-homosexual hostility in the country. PMID- 19296310 TI - Bisexual desire and familism: Latino/a bisexual young men and women in New York City. AB - Families are of critical importance for Latino communities in the USA. Familism - or the cultural value that weighs on interdependence between nuclear and extended family members for support, emotional connectedness, familial honour, loyalty and solidarity - has been demonstrated to reduce sexual health risks among heterosexual youth, yet this relationship has not been examined among Latino bisexual teenagers. In this study, we examined how familism shapes sexual decision making regarding behaviour and expressions of bisexuality among Latino youth. To accomplish this, we conducted 25 in-depth interviews and ethnographic observations among bisexual male and female youth (15-19 years of age) for nine months in New York City. We carried out a recurrent theme analysis together with the selection of case studies to illustrate key themes regarding familism and Latino teenage bisexuality. Findings suggest that bisexual Latino youth valued closeness to their families by maintaining family ties and seeking their emotional and material support. The negative consequence for those who wanted to keep their bisexuality private is the constant surveillance of the family network. Familism is a complex construct that has a strong potential for providing insights into sexual health practices of bisexual Latino youth. PMID- 19296313 TI - Issues in the use of visual supports to promote communication in individuals with autism spectrum disorder. AB - PURPOSE: Visual supports are widely used and generally regarded as an effective resource for intervention with individuals who function on the autism spectrum. More cross-contextual research into their efficacy is required. METHOD AND OUTCOMES: In this article, we selectively review the research literature around visual supports based on an original conceptual model that highlights their contribution in the interpersonal social and communicative milieu of classrooms, homes and other daily living contexts. Attention is drawn to a range of practical and research issues and challenges in the use of visual supports as well as evidence of their effectiveness in enhancing participation, learning and social membership in this population. CONCLUSIONS: Areas for further research relating to the introduction and use of visual supports with the autism spectrum disorder population are identified. PMID- 19296314 TI - An insider's view on person-centred rehabilitation: a case study. AB - PURPOSE: The author, a rehabilitation specialist of more than 30 years, presents her observations of the medical and rehabilitation services her husband received after a ruptured abdominal aortic aneurysm. Building on the person-centred approach, she identified several problems and describes theory-based strategies how she and her husband (patient) addressed these problems. In addition, she provides insight in the complex role she fulfilled as family caregiver: liaison, advocate and as personal health manager (taking care of her own health). METHOD: A qualitative analysis and interpretation based on personal observations and discussions with medical and rehabilitation professionals. RESULTS: The author argues that the culture of the institution (knowledge and compassion) and the role of the patient advocate have major implications on the outcome of a patient's functional gains. The International Classification of Functioning, Disability and Health can be a useful tool to enhance transparency within the patient-centred approach. CONCLUSION: Although this case study is described from the perspective of one case, the information shared in this paper may stimulate further discussion and research in the concept and implementation of person centred rehabilitation. PMID- 19296315 TI - Can younger drivers be trained to scan for information that will reduce their risk in roadway traffic scenarios that are hard to identify as hazardous? AB - Younger drivers (18-21 years) are over-involved in crashes. Research suggests that one of the reasons for this over-involvement is their failure to scan areas of the roadway for information about potential risks in situations that are hazardous, but not obviously so. The primary objective of the present study is to develop and evaluate a training program that addresses this failure. It was hypothesised that PC-based hazard anticipation training would increase the likelihood that younger drivers would scan for potential hazards on the open road. In order to test this hypothesis, 12 trained and 12 untrained drivers' eye movements were measured as they drove a vehicle on local residential, feeder and arterial roads. Overall, the trained drivers were significantly more likely to gaze at areas of the roadway that contained information relevant to the reduction of risks (64.4%) than were the untrained drivers (37.4%). Significant training effects were observed even in situations on the road that were quite different from those shown in training. These findings have clear implications for the type of training of teen drivers that is necessary in order to increase their anticipation of hazards. PMID- 19296316 TI - Multi-segment trunk kinematics during a loaded lifting task for elderly and young subjects. AB - The trunk is frequently modelled as one fixed segment ignoring possible multi segmental contributions during manual handling. This study compared segmental trunk motion in a young and older population during a lifting task. Twelve elderly and 19 young subjects repeatedly lifted a 5 kg box from bench to shelf under two stance conditions. Displacement and angular trunk segment kinematics were recorded with an electromagnetic tracker system and then analysed. The elderly subjects displayed significantly increased pelvic and trunk displacement and significantly reduced pelvic and lower thorax (T10-L1) range of motion in both stance conditions. Upper thorax (C7-T10) motion was at times greater than lumbar motion and opposite to the lower segments and was related to the task while the lower segments contributed to both equilibrium and task requirements. Decreased segmental trunk angular kinematics may contribute to increased displacement kinematics and place the elderly at increased risk of injury and falling. The pelvis, lumbar spine, low thorax (T10-L1), upper thorax (C7-10) contributed uniquely and synchronously to trunk (C7-S2) mechanics during a lifting task. Reduced angular kinematics of the pelvis and low thorax contributed to increased displacement kinematics and hence increased the risk of falling in the elderly compared to the young. Investigations of trunk mechanics should include multi-segment analysis. PMID- 19296317 TI - The Work Compatibility Improvement Framework: theory and application of improvement action and intervention strategies. AB - Challenges facing management of manufacturing firms can be transformed into asset gains by giving careful consideration to the worker-work environment interface. The benefits of a 'healthy' interface may lead to sizable reductions in rising health care costs and retention of highly qualified workers. This paper presents a novel approach for the 'improve' phase of the Work Compatibility Improvement Framework. The work tasks of this research consisted of: (a) fundamentals of cognitive-based improvement action and intervention; (b) design concepts and process of improvement action/intervention generation; (c) assessment model of estimated gains in company's assets; (d) application demonstration in the manufacturing sector. The process of improvement action/intervention generation is described, preceded by a description of the fundamentals of cognitive-based improvement action and intervention and system architecture. This is followed by a documentation of estimated asset gains as a result of the improvement plan. The results showed that expert workers were, on average, 78% in agreement with the algorithm-identified improvement actions. Their knowledge was used to update the recommended actions as well as to detail the multiple strategies required to address the improvement actions. As a result, an integrated improvement plan was developed resulting in estimated asset gains of $1.6 million, which was validated by the general manager. The research reported herein documented the theory and application of the 'improve' phase of the Work Compatibility Improvement Framework. The economic assessment of the suggested improvement is also reported and this has proved to be an important driver to secure the firm collaboration of manufacturing enterprise management. An integrated improvement solution plan backed by a detailed economic assessment of suggested improvements is essential to demonstrate the full potential of workplace micro- and macro-ergonomic interventions. PMID- 19296318 TI - The eye-complaint questionnaire in a visual display unit work environment: internal consistency and test-retest reliability. AB - The internal consistency and test-retest reliability of a 10-item eye-complaint questionnaire (ECQ) were examined within a sample of office workers. Repeated within-subjects measures were performed within a single day and over intervals of 1 and 7 d. Questionnaires were completed by 96 workers (70% female, mean age 36.4 years) who perform more than 4 h of visual display unit work each day. The internal consistency of the ECQ was high (alpha = 0.801). Although test-retest reliability was low within a single day (morning and afternoon) (intra-class correlation coefficient (ICC) = 0.68, 95% CI 0.38, 0.82), it was good between two morning measurements over a 1-d interval (ICC = 0.91 95% CI 0.87, 0.94) and between morning measurements with a 7-d interval (ICC = 0.87, 95% CI 0.81, 0.92). The ICC between afternoon measurements over a 1-week interval was moderate (ICC = 0.78, 95% CI 0.66, 0.85). This questionnaire is a reliable and consistent instrument for easily assessing eye complaints in office work. The timing of the measurements must be kept in consideration since measurements are confounded by exposures over the day. PMID- 19296319 TI - A longitudinal analysis of the effects of a preventive exercise programme on the factors that predict shoulder pain in construction apprentices. AB - Construction apprentices are at risk for developing shoulder pain with increasing exposure to repetitive overhead work. Risk may decrease if shoulder biomechanics are optimised and if risk factors that contribute to shoulder pain onset are identified. This prospective cohort study examined demographic and work-related factors and shoulder pain onset over 2 years in a cohort of 240 construction apprentices. Approximately 50% of the sample (n=117) performed a home exercise programme intended to have a protective effect, while the other 50% served as controls. The proportion of new-onset shoulder pain in the control group was higher than in the exercise group. Regression analysis identified four factors related to new-onset shoulder pain: previous neck pain; working in hot, cold or humid conditions; subject height; and bending and twisting the back. This information may assist employers and workers in preventing shoulder pain. By knowing factors predictive of shoulder pain development in construction workers, employers can take measures to protect workers and may secondarily decrease medical expenses and maintain productivity. Previous neck pain, working in extreme environmental conditions and being shorter all increased a worker's risk of developing shoulder pain. Exercises to optimise shoulder biomechanics have a small effect on preventing shoulder pain development. PMID- 19296320 TI - Effect of visual display height on modelled upper and lower cervical gravitational moment, muscle capacity and relative strain. AB - Head and neck posture is an important factor in neck pain related to computer use; however, the evidence for an optimal posture is unconvincing. This study measured the 3-D postures of 36 young adults during use of three different display heights. Cervical extensor muscle strain was estimated using modelled gravitational load moments and muscle capacities. The influence of more or less upper vs. lower cervical movement was also explored across a broad range of potential postures. Overall cervical extensor muscle capacity diminished away from a neutral posture whilst gravity moment increased with flexion. Overall cervical extensor muscle strain increased with head flexion but remained stable into head extension. Individual differences in the amount of upper and lower cervical movement had an important effect on strain, particularly for some muscles. Computer display height guidelines are an important component of ergonomics practice, yet the relative strain on neck extensor muscles as a function of display height has not been examined. The current findings provide more detailed biomechanical evidence that ergonomists can incorporate with usability and other evidence to determine appropriate display height recommendations. PMID- 19296321 TI - Workload demand in police officers during mountain bike patrols. AB - To the authors' knowledge this is the first paper that has used the training impulse (TRIMP) 'methodology' to calculate workload demand. It is believed that this is a promising method to calculate workload in a range of professions in order to understand the relationship between work demands and aerobic fitness. The aim of this study was to assess workload demand in police officers from the Utrecht police department in the Netherlands, during patrol by mountain bike. Maximum oxygen intake, maximum heart rate (HRmax), ventilatory threshold (VT)1 and VT2 were determined with a maximal exercise test on a bicycle ergometer. Heart rates were registered throughout three shifts in 20 subjects using a heart rate monitor. Exercise intensity was divided into three phases: phase I (between 40% of HRmax and VT1); phase II (between VT and the respiratory compensation point (RCP)); and phase III (>RCP). The total TRIMP score was obtained by summating the results of the three phases. Average daily workload demands of 355 TRIMPs per day and 1777 TRIMPs per week were measured. Workload demand approached and in some cases exceeded the upper limit of 2000 TRIMPs per week threshold level for physiological stress demands in professional male cyclists. PMID- 19296322 TI - Perceptual and physiological heat strain: examination in firefighters in laboratory- and field-based studies. AB - Firefighting demands performing heavy muscular work under adverse and potentially dangerous conditions. Although the physiological and psychological responses to simulated firefighting activities have been described, the heat strain has not been characterised using standardised indices of exercise-heat strain. The purpose of the study is to describe the physiological and perceptual strain associated with working in personal protective equipment and performing simulated firefighting activities in a hot environment using recently developed strain indices (Physiological Strain Index (PhSI); Perceptual Strain Index (PeSI)). Data from two previously published studies (Smith et al. 1995, 2001) - one a laboratory-based study and one a field-based study - were re-analysed incorporating the strain indices. The laboratory study involved walking on a treadmill for 15 min while wearing three different clothing and equipment configurations. The field study involved three trials of standardised firefighting tasks in a live-fire training structure (mean trial length = 5.76 min). Heart rate, rectal temperature, thermal sensations and ratings of perceived exertion were collected in each study. PhSI and PeSI values were calculated using the formulae developed by Moran et al. (1998b) and Tikuisis et al. (2002), respectively. PhSI and PeSI increased significantly over time in both studies. Even relatively brief bouts of exercise while wearing heavy impermeable clothing or simulated firefighting activity in the heat results in moderate to high levels of heat strain as assessed by PhSI and PeSI. PMID- 19296323 TI - Impact of automated decision aids on performance, operator behaviour and workload in a simulated supervisory control task. AB - In studies reporting automation effects on overall system performance and on the operator, the methods used to measure workload often did not appropriately reflect the complexity of this construct. The present study addresses the impact of automation on operator workload and behaviour in process control fault management. Workload effects were assessed with subjective, cardiovascular and secondary task performance indicators. Interactions with the interface of the process control simulation directed at gathering information and controlling the system were recorded. Automation made operators more efficient, allowing faster fault management with less information sampling and control actions. Subjective workload ratings were significantly lower in the assisted conditions as compared to manual, which was not reflected in cardiovascular and secondary task measures. Participants' information sampling activity did not differ between medium and high level of automation. Results suggest that participants paid constantly high attention to their task even with highly automated support. PMID- 19296324 TI - Lumbar spine forces during manoeuvring of ceiling-based and floor-based patient transfer devices. AB - Patient handling continues to represent a high risk task for low back pain (LBP) among health caregivers. Previous studies indicated that manual transfers of patients impose unacceptable loads on the spine even when two caregivers perform the transfer. Patient lift devices are considered a potential intervention; however, few biomechanical analyses have investigated the spine loads and LBP risk associated with these transfer devices. This study analysed the 3-D spine forces imposed upon the lumbar spine when 10 subjects manipulated ceiling-based and floor-based patient lifts through various patient handling conditions and manoeuvres. The results indicated that ceiling-mounted patient lift systems imposed spine forces upon the lumbar spine that would be considered safe, whereas floor-based patient handling systems had the potential to increase anterior/posterior shear forces to unacceptable levels during patient handling manoeuvres. Given these findings, ceiling-based lifts are preferable to floor based patient transfer systems. PMID- 19296325 TI - Effects of measurement errors on psychometric measurements in ergonomics studies: Implications for correlations, ANOVA, linear regression, factor analysis, and linear discriminant analysis. AB - This paper aims to demonstrate the effects of measurement errors on psychometric measurements in ergonomics studies. A variety of sources can cause random measurement errors in ergonomics studies and these errors can distort virtually every statistic computed and lead investigators to erroneous conclusions. The effects of measurement errors on five most widely used statistical analysis tools have been discussed and illustrated: correlation; ANOVA; linear regression; factor analysis; linear discriminant analysis. It has been shown that measurement errors can greatly attenuate correlations between variables, reduce statistical power of ANOVA, distort (overestimate, underestimate or even change the sign of) regression coefficients, underrate the explanation contributions of the most important factors in factor analysis and depreciate the significance of discriminant function and discrimination abilities of individual variables in discrimination analysis. The discussions will be restricted to subjective scales and survey methods and their reliability estimates. Other methods applied in ergonomics research, such as physical and electrophysiological measurements and chemical and biomedical analysis methods, also have issues of measurement errors, but they are beyond the scope of this paper. As there has been increasing interest in the development and testing of theories in ergonomics research, it has become very important for ergonomics researchers to understand the effects of measurement errors on their experiment results, which the authors believe is very critical to research progress in theory development and cumulative knowledge in the ergonomics field. PMID- 19296326 TI - Effects of speed-accuracy instructions on oculomotor scanning and target recognition in a simulated baggage X-ray screening task. AB - Visual search tasks are often carried out under high levels of time stress. Transportation security screeners, for example, face demands to achieve high levels of accuracy while maintaining rapid passenger throughput. An experiment examined the strategies by which operators regulate visual search performance under such conditions. Observers performed a simulated baggage-screening task under instructions to emphasise either response speed or accuracy. Behavioural measures and eye movements were recorded. Observers made fewer and briefer fixations under emphasise-speed than under emphasise-accuracy instructions. Losses in accuracy were produced by more frequent failures to fixate on targets and a decrease in the detection rate of non-fixated targets. The likelihood with which observers detected a fixated target was similar across speed-accuracy instructions. Results will inform efforts to model visual search in naturalistic tasks, allowing more accurate prediction of response times and error rate and may aid the design of training programmes and other interventions to improve search performance under stress. PMID- 19296327 TI - Upper limb discomfort profile due to intermittent isometric pronation torque at different postural combinations of the shoulder-arm system. AB - Twenty-seven right-handed male university students participated in this study, which comprised a full factorial model consisting of three forearm rotation angles (60% prone and supine and neutral range of motion), three elbow angles (45 degrees , 90 degrees and 135 degrees ), three upper arm angles (45 degrees flexion/extension and neutral), one exertion frequency (15 per min) and one level of pronation torque (20% maximum voluntary contraction (MVC) relative to MVC at each articulation). Discomfort rating after the end of each 5 min treatment was recorded on a visual analogue scale. Results of a repeated measures analysis of covariance on discomfort score, with torque endurance time as covariate, indicated that none of the factors was significant including torque endurance time (p = 0.153). An initial data collection phase preceded the main experiment in order to ensure that participants exerted exactly 20% MVC of the particular articulation. In this phase MVC pronation torque was measured at each articulation. The data revealed a significant forearm rotation angle effect (p = 0.001) and participant effect (p = 0.001). Of the two-way interactions, elbow*participant (p = 0.004), forearm*participant (p = 0.001) and upper arm*participant (p = 0.005) were the significant factors. Electromyographic activity of the pronator teres and biceps brachii muscles revealed no significant change in muscle activity in most of the articulations. Industrial jobs involving deviated upper arm postures are typical in industry but have a strong association with injury. Data from this study will enable better understanding of the effects of deviated upper arm postures on musculoskeletal disorders and can also be used to identify and control high-risk tasks in industry. PMID- 19296328 TI - Cardiac mitochondrial function and tissue remodelling are improved by a non antihypertensive dose of enalapril in spontaneously hypertensive rats. AB - Renal and cardiac benefits of renin-angiotensin system inhibition exceed blood pressure (BP) reduction and seem to involve mitochondrial function. It has been shown that RAS inhibition prevented mitochondrial dysfunction in spontaneously hypertensive rats (SHR) kidneys. Here, it is investigated whether a non antihypertensive enalapril dose protects cardiac tissue and mitochondria function. Three-month-old SHR received water containing enalapril (10 mg/kg/day, SHR+Enal) or no additions (SHR-C) for 5 months. Wistar-Kyoto rats (WKY) were normotensive controls. At month 5, BP was similar in SHR+Enal and SHR-C. In SHR+Enal and WKY, heart weight and myocardial fibrosis were lower than in SHR-C. Matrix metalloprotease-2 activity was lower in SHR+Enal with respect to SHR-C and WKY. In SHR+Enal and WKY, NADH/cytochrome c oxidoreductase activity, eNOS protein and activity and mtNOS activity were higher and Mn-SOD activity was lower than in SHR-C. In summary, enalapril at a non-antihypertensive dose prevented cardiac hypertrophy and modifies parameters of cardiac mitochondrial dysfunction in SHR. PMID- 19296329 TI - Medical, surgical and alternative treatments for chronic pelvic pain in women: a descriptive review. AB - Several causes of chronic pelvic pain (CPP) are recognised, but in many women a definite diagnosis cannot be made. Few randomised controlled trials on treatment of CPP have been conducted. In a Cochrane systematic review, only medroxy progesterone acetate, counselling, a multidisciplinary approach and lysis of deep adhesions had a proven benefit. The aim of this descriptive review is to describe the management of CPP, which can focus on treating the pain itself, the underlying cause, or both. Combination drug therapy with medications with different mechanisms of action may improve therapeutic results. Pelvic denervating procedures should be indicated in selected circumstances, as the magnitude of the effect is undefined. Several alternative non-invasive treatments have been proposed including exercise programmes, cognitive and behavioural medicine, physical therapy, dietary modification, massage and acupuncture. When the woman has completed her family and particularly when pelvic varices have been demonstrated, hysterectomy can be considered after a careful pre-operative assessment. However, substantial pain relief may be achieved in no more than 60 70% of the cases. A minority of patients (3-5%) will experience worsening of pain or will develop new symptoms after surgery. Treatment of CPP, generally, requires acceptance of the concept of managing rather than curing symptoms. PMID- 19296330 TI - Injury control & safety promotion. PMID- 19296331 TI - From myth to science in urban and transport planning: from uncontrolled to controlled and responsible urban development in transport planning. AB - Fossil energy use for mechanical transport modes enhanced travel speed far above human evolutionary experience, which is walking speed. Transport became faster and more convenient for people and industry. But planning had to be done without knowing the effects of these new modes. Individual experiences were extrapolated to the system and myths were created, like 'growth of mobility', 'time saving by speed' and 'freedom of modal choice'. Scientific based analysis show that these are real myths. These effects do not exist in the system. The number of trips is constant, travel time can not be saved in the system; speed lengthens distances and freedom of choice is limited by human evolution. Benefits from time saving can not be calculated any more and car traffic flow is only the effect of mistakes in parking organisation. PMID- 19296332 TI - Attitudes to seat belt wearing and related safety features in two cities in China. AB - The study aimed to establish characteristics of vehicles, occupants, seat belt and vehicle safety features for the Chinese cities of Nanjing and Zhoushan in order to interpret previously observed low seat belt wearing rates, particularly of passengers. Convenience interview surveys, stratified by vehicle and occupant type, of 2280 motor vehicle users were conducted in 2006 and 2007. Most participants (74.4%) were male, particularly drivers. Over half the occupants surveyed were aged 30-39 years. Drivers had relatively new vehicles and less driving experience compared with more motorised countries. Seat belts in most front seating positions and in half of the rear seats were fitted and functional. Where fitted, reasons for 'never wearing' were mostly 'feeling trapped and uncomfortable'; inconsistent wearers were influenced by 'habit development, trip length, speed, comfort and enforcement'. Public awareness of the safety benefits of seat belts, strong enforcement and retrofitting are necessary to overcome these barriers as achieved by past and present best practice. PMID- 19296333 TI - Injury deaths in the metropolitan region of Florianopolis, Southern Brazil, 1996 2002. AB - The objective was to analyse injury mortality in the metropolitan region of Florianopolis, Brazil, from 1996 to 2002. The study design was an ecological model. The mortality distribution was calculated along with the mortality rate due to injury in general and also by specific causes for each year during the study period and for the region's most populous towns. Injury mortality rates decreased in the period, but the homicide mortality increased by 103% when comparing the years 1996 and 2002. Most of the victims were children, young people and old people. This study confirmed a greater vulnerability among young people and the elderly, predominantly in the male population. The significant increase in the mortality rate due to homicides in this region was particularly noticeable and followed the same trend as has been observed in other regions of the country. PMID- 19296334 TI - Random or accidental--what is the goodness of the fit? PMID- 19296336 TI - A free software for the evaluation and comparison of dose response models in clinical radiotherapy (DORES). AB - PURPOSE: The aim of this work was to develop a user-friendly and simple tool for fast and accurate estimation of Normal Tissue Complication Probabilities (NTCP) for several radiobiological models, which can be used as a valuable complement to the clinical experience. MATERIALS AND METHODS: The software which has been named DORES (Dose Response Evaluation Software) has been developed in Visual Basic, and includes three NTCP models (Lyman-Kuther-Burman (LKB), Relative Seriality and Parallel). Required input information includes the Dose-Volume Histogram (DVH) for the Organs at Risk (OAR) of each treatment, the number of fractions and the total dose of therapy. RESULTS: NTCP values are computed, and subsequently placed in a spreadsheet file for further analysis. A Dose Response curve for every model is automatically generated. Every patient of the study population can be found on the curve since by definition their corresponding dose-response points fall exactly on the theoretical dose-response curve, when plotted on the same diagram. CONCLUSION: Distributions of absorbed dose alone do not provide information on the biological response of tissues to irradiation, so the use of this software may aid in the comparison of outcomes for different treatment plans or types of treatment, and also aid the evaluation of the sensitivity of different model predictions to uncertainties in parameter values. This was illustrated in a clinical case of breast cancer radiotherapy. PMID- 19296337 TI - Stress response of a p53 homologue in the radioresistant Sf9 insect cells. AB - PURPOSE: To investigate homology and stress response of p53 (a 53 kDa tumor suppressor protein) orthologue in Sf9 Lepidopteran insect cell line that exhibits very high radioresistance. MATERIALS AND METHODS: Western immunoblotting, immunoprecipitation, degenerate RT-PCR (reverse transcription-polymerase chain reaction), electrophoretic gel mobility shift assay, flow cytometry and immuno fluorescence microscopy were used for characterizing structural and functional features of Sfp53 (Spodoptera frugiperda p53) in gamma-irradiated or etoposide treated Sf9 insect and BMG-1 (brain malignant glioma) human cells. Cells were pre treated with caffeine for inhibiting ATM/ATR (ataxia-telangiectasia mutated protein/ATM and Rad-3-related protein) activation, wherever required. RESULTS: A 47-49 kDa protein band was observed with antibodies against three different epitopes, demonstrating conservation of respective domains in Sfp53. Immunoprecipitation also yielded similar-sized protein. Degenerate RT-PCR resulted in product of same size in both cell lines. Similar gel mobility shift of p53-binding oligonucleotide with BMG-1 and Sf9 cell lysates indicated analogous transcriptional activity of Sfp53. Constitutive Sfp53 level was higher than hp53 (human p53) and showed primarily cytoplasmic localization. Radiation induced accumulation was considerably less in Sf9 even as an analogous ATM/ATR dependent nuclear translocation was observed following gamma-irradiation and etoposide. CONCLUSIONS: A smaller-sized Sfp53 orthologue shows highly conserved native structure with DNA-binding, N-terminus and C-terminus domains, and has analogous p53 transcriptional activity. While its nuclear translocation and ATM/ATR dependence were similar to hp53, the cytoplasmic localization and subdued accumulation following gamma-irradiation indicate functional differences from human cells. PMID- 19296338 TI - Comparison of organic and inorganic germanium compounds in cellular radiosensitivity and preparation of germanium nanoparticles as a radiosensitizer. AB - PURPOSE: The aim of this work is to compare the radiosensitizing effect between organic and inorganic germanium compounds and to investigate whether nanometer sized germanium particles can act as radiosensitizers. MATERIALS AND METHODS: Bis (2-carboxyethylgermanium) sesquioxide (Ge-132), germanium oxide (GeO(2)) and germanium nanoparticles were used in this study. Cell viability was determined by clonogenic survival assay. Cellular DNA damage was evaluated by alkaline comet assay, confocal microscopy and the cellular level of phospho-histone H2AX (gamma H2AX). RESULTS: Nanometer-sized germanium particles were fabricated. They have a similar radiosensitizing effect as that of GeO(2). Conversely, Ge-132 did not enhance the radiosensitivity of cells. Comet assay was employed to evaluate the level of DNA damage and confirmed that inorganic germanium compounds enhanced cellular radiosensitivity. Notably, the comet assay indicated that the nanoparticle itself caused a higher level of DNA damage. The possibility that germanium nanoparticles per se caused DNA damage was ruled out when the cellular level of gamma-H2AX was examined. CONCLUSIONS: We demonstrated that inorganic but not organic germanium compounds exerted radiosensitizing effect in cells. Nanometer-sized germanium particles were fabricated and were able to enhance the radiosensitivity of cells. Confounding effect may occur when comet assay is used to estimate the level of DNA damage in the presence of germanium nanoparticles. PMID- 19296339 TI - Low-dose, low-dose-rate proton radiation modulates CD4(+) T cell gene expression. AB - PURPOSE: To evaluate cluster of differentiation 4(+) (CD4(+)) T cell gene expression and related parameters after whole-body exposure to proton radiation as it occurs in the spaceflight environment. MATERIALS AND METHODS: C57BL/6 mice were irradiated to total doses of 0, 0.01, 0.05, and 0.1 gray (Gy) at 0.1 cGy/h. On day 0 spleens were harvested from a subset in the 0, 0.01 and 0.1 Gy groups; (CD4(+)) T cells were isolated; and expression of 84 genes relevant to T helper (Th) cell function was determined using reverse transcriptase-polymerase chain reaction (RT-PCR). Remaining mice were euthanized on days 0, 4, and 21 for additional analyses. RESULTS: Genes with >2-fold difference and p < 0.05 compared to 0 Gy were noted. After 0.01 Gy, five genes were up-regulated (Ccr5, Cd40, Cebpb, Igsf6, Tnfsf4) and three were down-regulated (Il4ra, Mapk8, Nfkb1). After 0.1 Gy there were nine up-regulated genes (Ccr4, Cd40, Cebpb, Cxcr3, Socs5, Stat4, Tbx21, Tnfrsf4, Tnfsf4); none were down-regulated. On day 0 after 0.01 Gy, CD4(+) T cell counts and CD4:CD8 ratio were low in the spleen (p < 0.05). Spontaneous DNA synthesis in both spleen and blood was lowest in the 0.01 Gy group on day 0; on days 4 and 21 all p values were >0.1. CONCLUSION: The data show that the pattern of gene expression in CD4(+) T cells after protracted low dose proton irradiation was significantly modified and highly dependent upon total dose. The findings also suggest that low-dose radiation, especially 0.01 Gy, may enhance CD4(+) T cell responsiveness. PMID- 19296340 TI - Genetic damage in mammalian somatic cells exposed to extremely low frequency electro-magnetic fields: a meta-analysis of data from 87 publications (1990 2007). AB - PURPOSE: A meta-analysis was conducted to obtain a 'quantitative' estimate of the extent of genetic damage in mammalian somatic cells exposed to non-ionizing radiation emitted from extremely low frequency electro-magnetic fields (ELF-EMF) and to compare with that in unexposed control cells. METHODS: The methods used for the meta-analysis were recommended in several standard text books. Three specific variables related to ELF-EMF exposure characteristics were examined in the meta-analysis: (i) frequency (Hz), (ii) flux density (mT), and (iii) in occupationally exposed individuals. RESULT AND CONCLUSIONS: (1) The difference between ELF-EMF-exposed and control cells as well as the 'effect size' due to ELF EMF exposure were biologically small (although statistically significant) with very few exceptions. (2) At certain ELF-EMF exposure conditions there was a statistically significant increase in genetic damage assessed from some end points. (3) The mean indices for chromosomal aberrations and micronuclei end points in ELF-EMF-exposed and control cells were within the spontaneous levels reported in historical database. (4) Considerable evidence for publication bias was found in the meta-analysis. PMID- 19296341 TI - Cellular and sub-cellular responses to UVA in relation to carcinogenesis. AB - PURPOSE: UVA radiation (315-400 nm) contributes to skin aging and carcinogenesis. The aim of this review is to consider the mechanisms that underlie UVA-induced cellular damage, how this damage may be prevented or repaired and the signal transduction processes that are elicited in response to it. RESULTS: Exposure to ultraviolet (UV) light is well-established as the causative factor in skin cancer. Until recently, most work on the mechanisms that underlie skin carcinogenesis focused on shorter wavelength UVB radiation (280-315 nm), however in recent years there has been increased interest in the contribution made by UVA. UVA is able to cause a range of damage to cellular biomolecules including lipid peroxidation, oxidized protein and DNA damage, such as 8-oxoguanine and cyclobutane pyrimidine dimers. Such damage is strongly implicated in both cell death and malignant transformation and cells have a number of mechanisms in place to mitigate the effects of UVA exposure, including antioxidants, DNA repair, and stress signalling pathways. CONCLUSIONS: The past decade has seen a surge of interest in the biological effects of UVA exposure as its significance to the process of photo-carcinogenesis has become increasingly evident. However, unpicking the unique complexity of the cellular response to UVA, which is only now becoming apparent, will be a major challenge for the field of photobiology in the 21st century. PMID- 19296342 TI - Study on endocronological effects of radon speleotherapy on respiratory diseases. AB - BACKGROUND: Radon, as a radioactive noble gas of natural origin, is generally present in the atmosphere of caves during the speleotherapeutic treatment of different diseases. However, the role of radon in the therapeutic effect is not fully clarified yet. Endocrine parameter levels may be influenced by radon-like endocrine levels in balneotherapeutic treatments (bath treatment). For this reason changes of these parameters were examined in this study. PATIENTS AND METHOD: A total of 81 speleotherapy patients were involved in the survey. They spent four hours daily in Tapolca Cave, five days a week, for two weeks. In this study, the thyreoiedea stimulating hormone, free triiodine-tironin, free thyroxine, prolactine, cortisol, dehydroepiandrosterone sulphate, adrenocorticotropic hormone, and beta endorphine levels of the body were examined before and after the treatment. RESULTS: After statistical analysis of the data a significant decrease of cortisol levels of patients was found, which was not directly correlated with radon concentration. In the case of thyroid hormones, there were no significantly detectable changes of the hormone levels except for low radon concentration levels a significant decrease in the free thyroxine and the thyreoiedea stimulating hormone level of male patients was observed. CONCLUSION: Speleotherapy has an effect on the level of endocrine hormones; however, no direct correlation with differences in radon level was found. For patients whose endocrine levels fell outside the normal endocrine parameter values, more striking changes in endocrine hormone levels were seen, but changes of these parameters could not be statistically analyzed due to the small number of cases. PMID- 19296343 TI - Spontaneous frequency of micronuclei among the newborns from high level natural radiation areas of Kerala in the southwest coast of India. AB - PURPOSE: The present study was an attempt to determine the spontaneous frequency of micronuclei (MN) in newborns from High Level Natural Radiation Areas (HLNRA) and the adjoining Normal Level Radiation Areas (NLNRA) of the monazite-bearing Kerala Coast in Southern West India using Cytochalasin Blocked Micronuclei (CBMN) assay. MATERIALS AND METHODS: Human umbilical cord blood samples were collected from a total number of 271 newborns (61 from NLNRA and 210 from HLNRA), born to mothers aged between 17 and 37 years (mean maternal age: 24.08 +/- 4.23 years). Lymphocyte cultures were set up following microculture techniques and cultures were terminated at 72 hours. Cytochalasin B at a concentration of 4.0 microg/ml was added to the lymphocyte cultures at 44 h. Enumeration of micronuclei was restricted to Cytochalasin Blocked binucleated (BN) cells only. RESULTS: The frequency of MN among the newborns from NLNRA (1.40 +/- 0.12) per 1000 BN cells was not statistically significant as compared to HLNRA newborns (1.33 +/- 0.04) per 1000 BN cells. Our data did not show any radiation dose response. Odds ratios (OR) and confidence intervals (CI) have been calculated to see statistical significance in the mean MN frequency among the newborns from various dose groups with respect to control and it did not reveal any significant difference (p > 0.05). A marginal increase in the frequency of micronuclei was observed among the female newborns as compared to males with increasing mothers' age groups except for the mothers aged > 30 years, though not statistically significant. CONCLUSION: The baseline frequency of micronuclei in HLNRA newborns is not statistically different from NLNRA newborns suggesting that elevated level of naturally occurring radiation has no significant effect on the induction of micronuclei frequency among the newborns. PMID- 19296344 TI - Computational analysis of the number, area and density of gamma-H2AX foci in breast cancer cells exposed to (111)In-DTPA-hEGF or gamma-rays using Image-J software. AB - PURPOSE: To develop a simple method for the quantification of gamma-H2AX focus number, density and size. METHODS: MDA-MB-468 human breast cancer cells were treated overnight with (111)In-diethylenetriaminepentaacetic acid human epidermal growth factor ((111)In-DTPA-hEGF, 0-142 kBq/pmol) or exposed to gamma-radiation to induce DNA double strand breaks (DSB). DNA DSB formation was evaluated by detection of phosphorylated histone H2AX on serine 139 (gamma-H2AX) using immunofluorescence. Confocal microscopy was used to capture images of gamma-H2AX foci and cell nuclei. Image-J software with customized macros was used to quantify gamma-H2AX foci. RESULTS: The number of gamma-H2AX foci per nucleus scored using Image-J correlated strongly with the number scored using direct visual confirmation (coefficient of determination, R(2) = 0.950; 60 nuclei scored). The mean density (grayscale values per pixel), area and integrated density (IntDen) of individual foci increased linearly as the specific radioactivity (SR) increased up to 67 kBq/pmol (R(2) values of 0.826, 0.964, 0.978, respectively). The mean number of foci per nucleus, the combined area of gamma-H2AX foci per nucleus and the IntDen per nucleus also increased linearly with SR, giving R(2) values of 0.926, 0.974 and 0.983, respectively. Similar linear relationships were observed with the gamma-ray absorbed dose up to 3.0 Gy. CONCLUSIONS: The density, area and IntDen of individual foci, as well as the number of gamma-H2AX foci, total focus area and IntDen per nucleus were successfully quantified using Image-J with customized macros. PMID- 19296345 TI - Microscopic imaging of DNA repair foci in irradiated normal tissues. AB - PURPOSE: It is now feasible to detect DNA double strand breaks (DSB) in tissues by measuring the induction and resolution of DNA repair foci, such as gamma-H2AX, using immunofluorescent microscopy and digital image analysis. This review will highlight principal tools and approaches to tissue microscopy and analysis. It will also discuss the practical considerations of using microscopy in vitro and in vivo in measuring intranuclear foci following irradiation. CONCLUSIONS: Computer-based image analysis algorithms allow an objective and quantitative analysis of foci and protein-protein interactions using 3D confocal images. Finally, we review the literature in which DNA repair foci have been investigated as a biodosimeter or a biomarker of DNA repair in normal tissues. PMID- 19296346 TI - Fruits are rich in antioxidants and ripe for topical therapy. AB - BACKGROUND/AIMS: Human skin, exposed to a plethora of environmental insults that lead to free radicals and free radical damage, is equipped with an endogenous set of antioxidants, but these systems can become overwhelmed. Much study has focused on the therapeutic role of topical antioxidants but only a few investigate the utility of fruit extracts. Fruits are a rich source of diverse antioxidants and this article reviews their use for topical antioxidant protection. METHODS: MEDLINE searches from 1966 to August 2008 and EMBASE searches from January 1974 through August 2008 were conducted for articles in English. In vitro and in vivo studies of fruit extracts used specifically with skin tissue or skin cells were selected for review. CONCLUSIONS: Fruits show diverse antioxidant activity in vitro and upon topical use in vivo. Studies have focused on antioxidant activity against reactive oxygen species (ROS) and ROS-induced signaling cascades. However, there has been little study regarding activity against reactive nitrogen or carbonyl species. Fruits may serve as a versatile source of antioxidants that can be harnessed for topical use. Several studies have shown efficacy in human in vivo studies and future studies will elucidate the versatility and clinical use of fruit extracts. PMID- 19296347 TI - Addition of zinc and manganese to a biological dressing. AB - BACKGROUND: Zinc and manganese, which are used in vivo because of their healing properties, have been shown to modulate in vitro integrin expression and to enhance keratinocyte migration. In addition, at the clinical level, a dressing of keratinocytes suspended in a fibrin glue has been proposed for the treatment of chronic wounds. OBJECTIVE: To investigate whether the addition of trace elements to this dressing could modulate the migratory phenotype of keratinocytes via the modulation of integrin expression in a manner similar to an in vitro model and thus increase the healing properties of this dressing. METHODS: Keratinocytes were mixed with Tissucol and maintained in culture for 12 days in a medium either supplemented or not with zinc or manganese. Then, integrin expression was studied by immunohistochemistry on fibrin clot cryosections. RESULTS: We observed a significant increase of alpha5beta1 with zinc compared to the control medium. Zinc also enhanced alphaVbeta6 expression and manganese alpha5beta1, alphaVbeta5 and alphaVbeta6 expression, however without reaching a significant level. CONCLUSION: By modulating integrin expression, trace elements can improve the efficiency of a biological dressing made of keratinocytes in a fibrin glue matrix and, thus, it appears beneficial to add them to this biological dressing for the treatment of skin defects. PMID- 19296348 TI - Development of air sampling strategies for monitoring common air pollutants in a mission area at Camp Victoria in Kosovo--stationary vs. personal monitoring. AB - It can be problematic to collect representative samples in the occupational environment of deployed soldiers using personal samplers. In this study, several air sampling strategies were examined to identify the most efficient method for collecting air samples that were representative of the soldiers' exposure profiles at the Swedish KFOR Camp Victoria outside Pristina, Kosovo. Stationary monitoring was performed during two 5-day sampling campaigns, one in summer and one in winter. The acquired data were related to measurements obtained by personal monitoring of three and four subgroups, respectively. Patrolling soldiers, mechanics, and indoor workers were selected to represent groups with different exposure profiles, and the particles, metals associated with particles, nitrogen dioxide, sulfur dioxide, polycyclic aromatic hydrocarbons, aldehydes, and volatile organic compounds that they were exposed to were measured. Generally, low concentrations of the analytes were found in both sampling periods, but the variability was greater in the winter campaign. Samples collected by stationary samplers captured most of the variation in the personnel's exposure, according to principal component analysis (PCA). However, the results also indicate that personal exposure to most potential pollutants would be underestimated if a single outdoor station was used to monitor them. Nevertheless, combined data from stations located outdoors and in relevant microenvironments (workshop, office, and guard station) provided substantially better representation. Thus, it may be possible to obtain monitoring data without using personal samplers in these inherently problematic situations. PMID- 19296350 TI - Drug policy, politics, and the law. PMID- 19296351 TI - Medical marijuana: the conflict between scientific evidence and political ideology. Part one of two. AB - Whether "medical marijuana" (Cannabis sativa used to treat a wide variety of pathologic states) should be accorded the status of a legitimate pharmaceutical agent has long been a contentious issue. Is it a truly effective drug that is arbitrarily stigmatized by many and criminalized by the federal government? Or is it without any medical utility, its advocates hiding behind a screen of misplaced (or deliberately misleading) compassion for the ill? Should Congress repeal its declaration that smoked marijuana is without "current medical benefit"? Should cannabis be approved for medical use by a vote of the people as already has been done in 13 states? Or should medical marijuana be scientifically evaluated for safety and efficacy as any other new investigational drug? How do the competing- and sometimes antagonistic--roles of science, politics and prejudice affect society's attempts to answer this question? This article examines the legal, political, policy, and ethical problems raised by the recognition of medical marijuana by over one-fourth of our states although its use remains illegal under federal law. Although draconian punishment can be imposed for the "recreational" use of marijuana, I will not address the contentious question of whether to legalize or decriminalize the use of marijuana solely for its psychotropic effects, a fascinating and important area of law and policy that is outside the scope of this paper. Instead, the specific focus of this article will be on the conflict between the development of policies based on evidence obtained through the use of scientific methods and those grounded on ideological and political considerations that have repeatedly entered the longstanding debate regarding the legal status of medical marijuana. I will address a basic question: Should the approval of medical marijuana be governed by the same statute that applies to all other drugs or pharmaceutical agents, the Food, Drug, and Cosmetic Act (FD&C Act), after the appropriate regulatory agency, the Food and Drug Administration (FDA), has evaluated its safety and efficacy? If not, should medical marijuana be exempted from scientific review and, instead, be evaluated by the Congress, state legislatures, or popular vote? I will argue that advocacy is a poor substitute for dispassionate analysis, and that popular votes should not be allowed to trump scientific evidence in deciding whether or not marijuana is an appropriate pharmaceutical agent to use in modern medical practice. PMID- 19296352 TI - Ketorolac tromethamine transdermal gel: development, in vitro and in vivo evaluation. AB - The authors developed and evaluated a transdermal gel formulation of ketorolac tromethamine (ketorolac) for the treatment of nociceptive somatic pain. The formulation was optimized for skin permeation enhancers, pH of the system, and dosage strength using in vitro and in vivo techniques. Of the various permeation enhancers evaluated, dimethyl sulfoxide (DMSO) and oleic acid were found to significantly increase skin permeation flux of ketorolac. The concentration of DMSO affected the rate as well as extent of transdermal absorption. Use of citric acid further improved the skin penetration of ketorolac. In vitro diffusion results indicated significant increase in drug permeation with increasing drug concentration. However, the same did not translate into higher skin permeation during in vivo study. Although the area under concentration time curve (AUC(0-t)) increased significantly with increasing dose, the effect on maximum serum concentration (C(max)) was insignificant. The formulation can be used for inflammatory pain management while avoiding gastric adverse events associated with oral ketorolac. PMID- 19296353 TI - Agmatine levels in the cerebrospinal fluid of normal human volunteers. AB - Agmatine is an amine formed by the decarboxylation of l-arginine by the enzyme arginine decarboxylase. The fact that exogenous agmatine modulates morphine analgesia and dependence raises the question of whether the biosynthesis of endogenous agmatine is regulated during chronic pain. As a first step to understand the biological role of agmatine in human neurological and psychiatric conditions, this study was aimed to determine the levels of cerebrospinal fluid (CSF) agmatine in normal individuals. The levels of agmatine in the CSF and blood were measured by high-performance liquid chromatography (HPLC) method. Samples of CSF and blood were collected from a total of 10 participants for this study. The CSF agmatine levels ranged from 24.3 to 54.0 ng/mL, whereas the plasma agmatine levels were from 8.4 to 65.1 ng/mL. The mean values with standard error for blood and CSF agmatine were 33.8 +/- 16.6 and 40.4 +/- 9.1, respectively. The statistical analysis of these 10 samples indicated no correlation between blood and CSF samples (r = .29); however, removing one outlier improved the correlation (r = .6). From this study, the authors conclude that human CSF agmatine levels can be measured by HPLC with precision and that a possible correlation exists between plasma and CSF agmatine levels. This study provides basis for future studies in human chronic pain conditions. PMID- 19296354 TI - Nonsteroidal anti-inflammatory drug usage and gastrointestinal outcomes in the Republic of Serbia. AB - This study assessed the utilization of prescription and over-the-counter (OTC) nonsteroidal anti-inflammatory drugs (NSAIDs) as well as the rate of self medication with diclofenac, ibuprofen, and naproxen in the Braneechevo District of the Republic of Serbia. Estimation of gastrointestinal (GI) disease (morbidity) trends and GI toxicity-associated hospitalization were studied and direct costs due to NSAID-induced GI toxicity are presented. This descriptive, retrospective study addressed drug use and outcomes between 2004 and 2006 documented in the Health Insurance Fund database of the Pozarevac Public Pharmacy in the Pozarevac Public Health Centar of the Braneechevo District, which includes 200,503 inhabitants. Data type/selection were defined daily doses (DDD) per 1000 inhabitants per day for utilization of drugs, number of patients with ICD-9 diagnosis codes for GI disorders; GI hospitalization count (average annually length of stay [in days] and number of GI hospitalizations); direct cost of hospital care. The OTC diclofenac use showed an increasing tendency: 6.2279; 6.5983; 8.2911 DDD units, as well as the utilization of OTC ibuprofen: 2.4389, 2.4899, 2.5776 DDD units, respectively (2004-2006), whereas OTC naproxen had relatively low utilization. In the same period, GI morbidity decreased: 9636, 7982, 7806, respectively, and the number of GI morbidity-associated hospitalizations increased 10.18% in 2005 and 15.06%, in versus 2004. The costs of GI morbidity-associated hospitalizations increased: 12.20% (2005) and 94.51% (2006), compared to 2004 costs with a positive correlation between utility of diclofenac and ibuprofen (self medication) and increased GI hospitalizations in Braneechevo. PMID- 19296355 TI - Evidence-based medicine works best when there is evidence: challenges in palliative medicine when randomized controlled trials are not possible. AB - Randomized controlled trials provide the best evidence for the safety and effectiveness of interventions. However, the reliance on this study design as the only criterion for credible evidence is not possible in all areas of palliative medicine. Although we must exercise caution in making definitive claims from uncontrolled trial data, limitations in performing controlled trials should neither preclude the use of findings from well-designed nonrandomized controlled trials nor, more importantly, deprive patients of potentially effective treatments. PMID- 19296356 TI - Patient education and self-advocacy: questions and responses on pain management. PMID- 19296357 TI - Pain and palliative care pharmacotherapy literature summaries and analyses. AB - Timely and important studies are reviewed and commentaries provided by leading palliative care clinicians. Symptoms, interventions, and treatment-related adverse events addressed in this issue are management of Alzheimer's agitation with donepezil; needle-free lidocaine powder for minor painful procedures; psychostimulants in depression; anticoagulation for cancer-related venous thromboembolism; effect of waiting for acute pain treatment on risk of chronic pain; and an update on severe cutaneous reactions associated with medications. PMID- 19296358 TI - Evidence-based pain management and palliative care systematic reviews in issue three for 2008 of The Cochrane Library. AB - The Cochrane Library of Systematic Reviews is published quarterly. Issue Three for 2008 contains 3539 complete reviews, 1868 protocols for reviews in production, and 8349 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 540,000 randomized controlled trials, and 10,000 cited papers in the Cochrane methodology register. The health technology assessment database contains 7400 citations. This edition of the Library contains 75 new reviews, of which 7 have potential relevance for practitioners in pain and palliative medicine. PMID- 19296359 TI - International perspectives on pain and palliative care. PMID- 19296362 TI - Differentiation of power and endurance athletes based on their muscle fatigability assessed by new spectral electromyographic indices. AB - The aim of this study was to differentiate between endurance and power athletes based on electromyographic (EMG) data analysed using new spectral indices. Nine endurance and six strength athletes were recruited to complete sets of knee extension repetitions (15 per set) until exhaustion, with each set followed by a maximal voluntary isometric knee extensor contraction. Peripheral muscle fatigue of the vastus lateralis, vastus medialis, and rectus femoris (bilaterally) was quantified by the changes in median frequency of the EMG power spectrum and a new spectral EMG fatigue index. Cluster analysis of the fatigue indices differentiated athletes into two groups: endurance (fatigue resistant) and strength (faster fatigue), whereas cluster analysis of the median EMG power spectrum frequency produced six indistinct groups. The average fatigue index for the quadriceps group increased across repetitions by 40 +/- 24% in the endurance group and by 184 +/- 12% in the strength group. The decrease in peak force and power across repetitions, and the rate of force decrease during maximal voluntary contraction per set, were significantly smaller for the endurance than for the strength group. The new spectral EMG indices effectively discriminated between strength and endurance athletes, thus providing a useful functional index that could be applied to track training adaptations as well as potentially talent identification. PMID- 19296363 TI - Haemodynamic changes after prolonged water immersion. AB - Thermoneutral water immersion increases cardiac preload and changes the neuroendocrine settings of blood volume regulation. The resulting marked diuresis may lead to significant haemodynamic changes after the end of a prolonged water immersion. Ten volunteers underwent 6 h of complete thermoneutral water immersion. Changes in cardiovascular status were assessed 1 h and 16 h after water immersion. Haemodynamic changes were assessed by Doppler echocardiography. Arterial wall distensibility was estimated by pulse wave velocity analysis. One hour after water immersion, mean weight loss was 1.78 kg and urine volume amounted to 1.5 litres. Echocardiographic measurements evidenced a significant decrease in dimensions of the left cardiac chambers and inferior vena cava. The decreased cardiac preload was paralleled by a lower stroke volume and cardiac output. A peripheral vasoconstriction associated with a relative decrease in the lower limb blood flow was evidenced by an increase in carotid-pedal pulse wave velocity and by a decrease in ankle brachial index. Sixteen hours after water immersion, cardiac preload and cardiac output remained below baseline values and peripheral vascular tone was still higher than at baseline. Marked haemodynamic changes had not returned to baseline 16 h after water immersion. There is a need to design fluid-replacement protocols to improve this recovery. PMID- 19296364 TI - Adjustment to change in familiar and unfamiliar task constraints. AB - The aim of this study was to assess the rate of adjustment to changes in task constraints that are familiar and unfamiliar when a change in the pattern of sequencing of segmental movements is not required. The selected task was underwater flutter kicking with flippers (familiar) and without flippers (unfamiliar). Nine male competitive age-group swimmers were assigned either to an unfamiliar "leg flipper" task, a familiar "foot flipper" task or a "control" group to perform five trials without flippers (all groups), 60 trials with flippers, and a post-test comprising 10 trials without flippers (all groups). Kinematic variables were calculated from digitized video data. Whether the movement pattern was appropriate for the task was indicated by the percent power in the fundamental Fourier frequency harmonic of the vertical oscillations of the hip, knee, and ankle, and by a "velocity index" that was defined as the ratio of the hip-to-knee and knee-to-ankle velocities of the fundamental Fourier waveform. Adjustment to reproduce the appropriate movement pattern occurred within the first block of 10 trials regardless of whether the constraints were familiar or unfamiliar. However, optimal performance in terms of swimming speed was not obtained following change to the unfamiliar constraint until after 10 trials. PMID- 19296365 TI - Side biases for holding and carrying infants: Reports from the past and possible lessons for today. AB - Most adults hold human infants on the left side, with the infant's head to the left of their own body midline. The discovery of this bias is credited to Lee Salk, who first reported it in 1960, but the same was reported at least 300 years earlier and many times again through the early decades of the twentieth century. Along with the left-side reports, however, others named the right as the preferred side. Authors on each side explained the preference and foresaw consequences for the infant: different ones in each case. This article describes the two kinds of reports, asks whether and how they might be reconciled, and discusses their possible lessons for theory and research today. PMID- 19296366 TI - In vitro activity of nine antifungal agents against clinical isolates of Aspergillus calidoustus. AB - This study analyzes Aspergillus isolates from the Spanish National Centre for Microbiology collection, which were identified morphologically as members of Aspergillus section Usti. Strains were identified through the analysis of the Internal Transcribed Spacer regions and partial beta tubulin gene sequences. One Aspergillus pseudodeflectus isolate and eight Aspergillus calidoustus isolates were detected in this panel of clinical strains. Terbinafine and the echinocandins micafungin and anidulafungin, were the drugs most active against these species. PMID- 19296367 TI - Invasive aspergillosis in chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is an inherited disorder of the NADPH oxidase complex in which phagocytes are defective in generating superoxide anion and its metabolites. NADPH oxidase activation leads to activation of sequestered neutrophil proteases that mediate host defense. Invasive aspergillosis and other rarer mold diseases are the leading causes of mortality in CGD, reflecting the key role of the phagocyte NADPH oxidase in host defense against opportunistic fungi. Despite recombinant interferon-gamma prophylaxis, invasive filamentous fungal infections are a persistent problem in CGD. Key principles of management of fungal infections involve early recognition and aggressive treatment and appropriate surgical debridement of localized disease. Because CGD is a disorder of phagocyte stem cells in which the gene defects are well defined, it is a model disease to evaluate immune reconstitution through stem cell transplantation and gene therapy. Recent studies using CGD mice show that defects in tryptophan catabolism may underlie the impaired host defense and pathogenic inflammation in CGD and open the potential for novel therapeutic approaches; however, correlative studies in patients are required. PMID- 19296368 TI - Echinocandin activity against Aspergillus spp. and the importance of pharmacodynamics. AB - Echinocandins represent a safe and well tolerated option for the therapy of invasive aspergillosis in patients who are unable to tolerate other agents or have refractory disease. In vitro and animal model data provide useful insights into the activity, appropriate dose, and potential role of these agents in invasive aspergillosis. These studies reveal a potentially concerning lack of tissue sterilization when echinocandins are used as monotherapy but clearly show improved survival with increasing doses. Furthermore pharmacodynamic studies suggest that echinocandin doses currently in widespread clinical use may optimize outcomes in invasive aspergillosis. A paucity of clinical data exists examining these agents as monotherapy for invasive aspergillosis and virtually no clinical data exists for using these agents as primary therapy. Further data examining the role of echinocandin monotherapy for invasive aspergillosis is unlikely to be forthcoming in the foreseeable future due to several factors including the aforementioned issues, the relatively small number patients with this infection, and the belief that these agents are potentially best used as part of combination therapy regimens for invasive aspergillosis. PMID- 19296369 TI - Sex, drugs, and rock and roll-teaching with adolescent standardized patients. PMID- 19296370 TI - 'So you want to be a clinician-educator...': designing a clinician-educator curriculum for internal medicine residents. AB - BACKGROUND: Despite a growing demand for skilled teachers and administrators in graduate medical education, clinician-educator tracks for residents are rare and though some institutions offer 'resident-as-teacher' programs to assist residents in developing teaching skills, the need exists to expand training opportunities in this area. METHODS: The authors conducted a workshop at a national meeting to develop a description of essential components of a training pathway for internal medicine residents. Through open discussion and small group work, participants defined the various roles of clinician-educators and described goals, training opportunities, assessment and resource needs for such a program. RESULTS: Workshop participants posited that the clinician-educator has several roles to fulfill beyond that of clinician, including those of teacher, curriculum developer, administrator and scholar. A pathway for residents aspiring to become clinician educators must offer structured training in each of these four areas to empower residents to effectively practice clinical education. In addition, the creation of such a track requires securing time and resources to support resident learning experiences and formal faculty development programs to support institutional mentors and leaders. CONCLUSION: This article provides a framework by which leaders in medical education can begin to prepare current trainees interested in careers as clinician-educators. PMID- 19296371 TI - In vitro synergistic antibacterial activities of helvolic acid on multi-drug resistant Staphylococcus aureus. AB - An agar plate method was established to screen synergistic antibacterial agents other than beta-lactamase inhibitors. By using this method, a strain Aspergillus sp136 was selected for further studies. From the metabolites of this strain, a synergistic antibacterial compound was isolated by bioautographic TLC assay guided fractionation and identified as helvolic acid. The synergistic effect of helvolic acid to penicillin was about 3 times that of clavulanic acid to penicillin in agar diffusion assay on Bacillus cereus. In checkerboard studies, helvolic acid exhibited synergistic effects with erythromycin on all tested multi drug resistant Staphylococcus aureus and with penicillin and tetracycline on some multi-drug resistant S. aureus. A pattern of enhanced killing was also found in time-kill studies on multi-drug resistant S. aureus. PMID- 19296372 TI - In vitro and in vivo effects of clove on pro-inflammatory cytokines production by macrophages. AB - Biological properties of clove have been reported, but little is known about its effect on the immune system. This work was aimed to investigate the effect in vivo of a water-soluble part of hydroalcoholic extract of clove on pro inflammatory cytokines (IL-1beta and IL-6) production by macrophages of BALB/c mice. The action of the essential oil of clove on the production of these cytokines macrophages was also investigated in vitro. The chemical compositions of the extract and of the oil were also investigated. Treatment of mice with water extract of clove was found to inhibit macrophages to produce both IL-1beta and IL-6. The essential oil of clove also inhibited the production of these cytokines in vitro. Eugenol was found to be the major component of the clove extract and essential oil, and probably is the causative agent of cytokine inhibition. Taken together, these data suggest an anti-inflammatory action of this spice. PMID- 19296373 TI - Antinociceptive activity of Gynandropsis gynandra leaves. AB - Gynandropsis gynandra (Capparidaceae) leaves are traditionally used in the treatment of pain. In order to evaluate the scientific validity of this, leaves were extracted successively to produce various extracts. These extracts were screened for antinociceptive activity using the hot plate test and acetic acid induced writhing test in mice at a dose of 100 mg kg(-1), intraperitoneally. Ethanol and aqueous extracts were found most active in both the tests. The action was blocked by naloxone (1 mg kg(-1), s.c.) in the hot plate test, which suggests involvement of opioid receptors in the action. Flavonoids and tannins were observed in the active extracts, so we can say that they may responsible for the antinociceptive activity. PMID- 19296374 TI - Studies on the anti-psoriasis constituents of Oplopanax elatus Nakai. AB - Seven compounds were isolated from roots and stems of Oplopanax elatus, of which compounds 3, 4, 5 and 6 were isolated for the first time from the title plant; compounds 1 and 2 are new compounds and characterised to be 3,3'-dimethoxy-4,9,9' trihydroxy-4',7-epoxy-5'8-lignan-4,9-bis-O-beta-D-glucopyranoside and 5 methoxylariciresinol-4-O-beta-D-glucopyranoside on the basis of NMR spectra and CD spectrum. PMID- 19296375 TI - Essential oil composition and variability of Laurus nobilis L. growing in Tunisia, comparison and chemometric investigation of different plant organs. AB - Stems, leaves, buds and flowers of Laurus nobilis L. growing wild in Tunisia were analysed for their essential oil composition. The essential oil of Laurus nobilis L. gathered from different stations were isolated by hydrodistillation and analysed by GC/MS. The oil yields on a dry weight basis ranged between 0.4% and 1.1%. The major component identified was 1,8-cineole, other predominant components were alpha-terpinyl acetate, methyl eugenol, eugenol and linalool. Although the same compounds were present in all plant organs, the leaves differed from the stems in the concentration of 1,8-cineole and methyl eugenol, buds and flowers in the concentration of 1,8-cineole and the stem's oil composition differs from the others in content of methyl eugenol. The results obtained from GC/MS analysis of the volatile oils from individual plant organs were submitted to principal component analysis. Chemometric investigations led to differentiation of stems, leaves and buds-flowers with the respect to the content of 1,8-cineole, metyhyl eugenol and alpha-terpynil acetate; flowers and buds were non-differentiated. Finally, the antibacterial activity of the leaves' essential oils has been assayed. PMID- 19296376 TI - Cholinesterase inhibitors from Sargassum and Gracilaria gracilis: seaweeds inhabiting South Indian coastal areas (Hare Island, Gulf of Mannar). AB - Dementia is a chronic progressive mental disorder, which adversely affects memory, thinking, comprehension, calculation and language. Some of the commonest forms of dementia are Alzheimer's disease, Parkinsonism, Dementia with Lewy Bodies and Myasthenia gravis. All these disorders are related to abnormalities in the central cholinergic system, which shows a decline in acetylcholine level. Cholinesterase (ChE) inhibitors are one of the novel strategies used for the symptomatic treatment of neurological disorders like dementia. In the course of screening new ChE inhibitors from marine sources, about 11 seaweeds, which have wide pharmaceutical applications, were collected from Hare Island, Gulf of Mannar, Tamilnadu, India. Methanolic extracts of the seaweeds were assessed for ChE inhibitory activity under in vitro conditions. Kinetic parameters IC(50), K(i) and V(max) were also analysed. The results showed that 3/11 seaweeds showed 50% inhibition for both ChEs (using acetylthiocholine iodide and butyrylthiocholine iodide as substrate) at concentrations of 2 mg mL(-1) (Gracilaria gracilis, Sargassum, Cladophora fasicularis for ChE with acetylthiocholine iodide as substrate and Gracilaria gracilis, Gracilaria edulis, Sargassum for ChE with butyrylthiocholine iodide as substrate) and 4/11 showed no inhibitory activity. Inhibitory activity of seaweed extracts was compared with standard drug donepezil. Enzyme kinetic analysis showed that algal extracts exhibited mixed type inhibition (partially non-competitive inhibition). PMID- 19296377 TI - Biological activity of secondary metabolites from Peltostigma guatemalense. AB - Leaves and wood of Peltostigma guatemalense, a novel species of the family Rutaceae, yielded a total of 14 secondary metabolites, i.e. methyl p-hydroxy benzoate, phenylacetic acid, beta-sitosterol, lupeol, syringaresinol, scopoletin, gardenin B (1), and seven alkaloids: gamma-fagarine (2), skimmianine (3), kokusaginine (4), 7-O-isopentenyl-gamma-fagarine (5), anhydro-evoxine (6), evoxine (7) and 4-methoxy-1-methyl-quinolin-2-one (8). The compounds have been identified by spectroscopic methods. Antibacterial and antimalarial in vitro activity of the isolated compounds were also determined. Methyl p-hydroxy benzoate and quinolone (8) were the most effective on Plasmodium falciparium strains. PMID- 19296378 TI - Antioxidative activity of glycoprotein isolated from Geranium sibiricum Linne. AB - Glycoprotein from Geranium sibiricum Linne (GSL) with 18 kDa was isolated and it consists of carbohydrate moiety (10.45%) and protein moiety (89.55%). The GSL glycoprotein was characterised by its radical scavenging activity under various experimental conditions. When GSL glycoprotein was treated with deactivation agents (pronase E or NaIO(4)), its scavenging activity decreased in both cases. It has optimal and maximal activity in acidic, neutral pH (up to pH 9), and up to 85 degrees C. Also, its activity reduced in the case of Ca(2+) and Mn(2+), with the exception of the Mg(2+) case. Its activity in the presence of Mn(2+) declined more than in the case of the Ca(2+). Also, GSL glycoprotein (500 microg mL(-1)) has antioxidative effects on hydroxyl and superoxide anion radicals in cell-free systems, and GSL glycoprotein (200 microg mL(-1)) significantly protected from cytotoxicity in the GO (100 mU mL(-1))-treated Chang liver cells for 4 h. PMID- 19296379 TI - A new bioactive xanthanolide from Xanthium cavanillesii. AB - Development of new antimicrobial compounds against different microorganisms is becoming critically important, as infectious diseases are still one of the leading causes of death in the world. The pharmaceutical industry is searching for new lead compounds with novel chemical structures to overcome the increasing resistance to known antibiotics. Plants can be a useful source of these lead compounds. Xanthium cavanillesii Schouw, Asteraceae, grows wild in Uruguay and its infusion is used in traditional medicine as a skin antiseptic. We have previously reported studies on the antimicrobial activity of several extracts of X. cavanillesii against different microorganisms. In this work, we present the isolation and structural elucidation by spectroscopical methods of a sesquiterpene lactone with a new xanthanolide derived skeleton. PMID- 19296380 TI - Bioactive compounds of inhibiting xanthine oxidase from Selaginella labordei. AB - Four flavone compounds were isolated from the effective fractions inhibiting xanthine oxidase (XOD) of the medicinal plant Selaginella labordei with anti virus activity, and the structures were elucidated as 4'-methylether robustaflavone (1), robustaflavone (2), eriodictyol (3) and amentoflavone (4). The 50% inhibitory concentration (IC(50)) of the three compounds of inhibiting XOD were 61.0, 0.199, 16.0 and 32.0 mg L(-1), respectively. All of these compounds were isolated from the species for the first time, and eriodictyol was found from Selaginellaceae for the first time. Among these compounds, robustaflavone has been reported as an effective compound against the hepatitis B virus. PMID- 19296381 TI - Hypoglycaemic and antioxidant activities of Ficus racemosa Linn. fruits. AB - The effects of Ficus racemosa Linn. fruit extract and fraction on fasting serum glucose levels of normal, type 1 and type 2 diabetic model rats are presented. The aqueous 80% EtOH extract and its water soluble fraction of F. racemosa fruit did not show any serum glucose lowering effect on non-diabetic and type 2 diabetic rats at the fasting condition, whereas the extract showed significant hypoglycaemic effect on the type 1 diabetic model rats. Both the extract and fraction were consistently active in both non-diabetic and types 1 and 2 diabetic model rats when fed simultaneously with glucose load. On the contrary, they were ineffective in lowering blood glucose levels when fed 30 min prior to glucose load. The 1-BuOH soluble part of the ethanol extract exhibited significant antioxidant activity in DPPH free radical scavenging assay. 3-O-(E)-Caffeoyl quinate (1) was isolated for the first time from this plant, which also showed significant antioxidant activity. PMID- 19296382 TI - Elaeagnoside, chymotrypsin inhibiting steroidal glucoside from Elaeagnus orientalis. AB - Elaeagnoside (1), a new steroidal glucoside, has been isolated from the chloroform fraction of Elaeagnus orientalis along with beta-sitosterol and 12 hydroxy-8,10-octadecadienoic acid. The structure of 1 has been elucidated with the help of chemical and spectral studies. It showed significant inhibitory activity against the enzyme chymotrypsin. PMID- 19296383 TI - Alteration study of lipids and phospholipids compositions of Pachysolen tannophilus membrane with aqueous ethanol media. AB - The variations in lipid compositions of Pachysolen tannophilus membrane from aqueous to 5% aqueous ethanol solutions are examined. The decreases in phospholipids, sterols and unsaturated fatty acid contents and slight increase in glycolipids content are reported. PMID- 19296384 TI - Lignan derivatives from the stem bark of Syzygium cumini (L.) Skeels. AB - Phytochemical investigation of the stem bark of Syzygium cumini (L.) Skeels (Myrtaceae) yielded four new lignan derivatives characterised as (7alpha,8alpha,2'alpha)-3,4,5-trimethoxy-7,3',1',9'-diepoxylignan (cuminiresinol), (7alpha,7'alpha,8alpha,8'alpha)-3,4-dioxymethylene-3',4' dimethoxy-7,9',7',9-diepoxylignan-5'-ol (5'-hydroxy-methyl-piperitol), (7alpha,7'alpha,8alpha,8'alpha)-3'-methoxy-9-oxo-7,9',7',9-diepoxylignan-3,4,4' triol or 3-demethyl-9-oxo-pinoresinol (syzygiresinol A), (7alpha,7'alpha,8alpha,8'alpha)-9-oxo-7,9',7',9-diepoxylignan-3,4,3',4',5' pentaol or 3,3'-didemethyl-9-oxo-pinoresinol (syzygiresinol B) along with the known lignans di-demethyl-5-hydroxypinoresinol, dimethylpinoresinol, didemethoxypinoresinol, pinoresinol and 4'-methyl-5'-hydroxypinoresinol. The structures of these lignans were elucidated on the basis of structural data analysis and chemical reactions. PMID- 19296385 TI - Iridoids from Euphrasia genargentea, a rare Sardinian endemism. AB - The phytochemical study of Euphrasia genargentea, a rare species only present in Sardinia, led to the identification of iridoid glucosides, i.e. aucubin, catalpol, mussaenosidic acid and melampyroside, which allowed chemotaxonomic considerations on the genus. On the basis of iridoid distribution in the genus, E. genargentea does not show any particular analogy with other Italian Euphrasia spp. This study is also important considering the severe risk of extinction of E. genargentea. PMID- 19296386 TI - Steroidal constituents from the aerial parts of Lippia nodiflora Linn. AB - The methanolic extract of the aerial parts of Lippia nodiflora Linn. afforded one new and two known steroids. The structure of the new steroid had been elucidated as 4',5'-dimethoxybenzoloxystigmasterol (1) by an analysis of the spectral data, including extensive NMR experiments and its chemical transformation to stigmasterol. The known steroids were identified as stigmasterol and beta sitosterol through comparison of their spectral data with those reported in the literature. PMID- 19296387 TI - A novel flavanone from Ilex hainanensis Merr. AB - Chemical investigation of the leaves of Ilex hainanensis Merr. yielded a new flavanone named Huazhongilexone-7-O-beta-D-glucopyranoside, together with five known compounds, which included caffeic acid, quercetin, isoquercitrin, quercitrin-7-O-beta-D-glucopyranoside and rutin. All of the compounds were isolated from the plant for the first time. The structure of the new compound was determined on the basis of high resolution mass, 1D and 2D NMR spectroscopies including ROESY. The antioxidant activity of the new compound was assessed using the DPPH spectrophotometric and IC(50) value was 10.39 micromol L(-1). PMID- 19296388 TI - Chemical composition of the halophyte plant Stachys maritima Gouan from the Bulgarian Black Sea coast. AB - Four groups of metabolites from the halophyte plant Stachys maritima Gouan, family Lamiaceae, were investigated: lipids, sterols, volatiles and polar compounds. Most of the metabolites have been previously reported to possess biological activity. Two of the main substances in the volatile fraction- phenidone and naphthalene, unusual for plants--were products of environmental contamination. This is a typical example of how marine pollutants could transfer and threaten terrestrial organisms via the trophic chain. The n-butanol extract possessed a relatively high antibacterial activity against S. aureus and Candida albicans, but was not active against Escherichia coli. PMID- 19296389 TI - Saturated long-chain hydrocarbons from Annona squamosa L. bark. AB - The unsaponified chloroform extract of Annona squamosa L. bark has been analysed by gas chromatography-mass spectrometry (GC-MS) and was shown to consist of the conventional, even carbon number dominant distribution of saturated n-alkane homologues (C(14)-C(26)). The heptadecane (5.75%) was the only odd carbon number hydrocarbon. The total amount of hydrocarbons found in the unsaponified chloroform extract was 24.60%. PMID- 19296390 TI - Loliolide in marine algae. AB - Loliolide content was determined in 13 marine algae including red, brown and green algae collected from the Black Sea, the Dardanelles and the Aegean Sea. Identification and quantification were performed by gas chromatography-mass spectrometry. The loliolide content in green alga is 1.76 microg g(-1), ranges from 0.14 to 4.35 microg g(-1) in red and from 0.18 to 4.83 microg g(-1) in brown algae. The results obtained are in the same range as previously reported for algae, as well as terrestrial plants. This article represents the first report of loliolide occurrence in green algae. PMID- 19296391 TI - Phytochemical re-investigation of Gentiana utriculosa. AB - Xanthone-O-glycosides with 1,3,7,8-oxidation pattern and flavone isoorientin-3'-O glucoside were isolated from the aerial parts of Gentiana utriculosa. Xanthone-C glucoside lancerin was detected in Gentiana species for the first time. The distribution of these compounds within the section Calathianae is discussed. PMID- 19296392 TI - Three new insecticidal sesquiterpene pyridine alkaloids from Celastrus angulatus. AB - Three new sesquiterpene pyridine alkaloids with a beta-dihydroagarofuran sesquiterpene skeleton, celangulatin G(1), H(2) and I(3), and one known alkaloid 4 were isolated from the middle-polar extracts of the root bark of Celastrus angulatus by bioassay-guided separation. Their chemical structures were elucidated mainly by the analyses of the MS and NMR spectral data. Celangulatin G(1), H(2), I(3) and 4 showed KD(50) against Mythimna separata were 849.61, 80.24, 96.29 and 50.98 microg g(-1), respectively. PMID- 19296393 TI - Leishmanicidal potential of N-substituted morpholine derivatives: synthesis and structure-activity relationships. AB - A series of N-substituted morpholines 2-20 was synthesised by reacting various acid chlorides and alkyl halides with morpholine (1). All of the synthesised compounds 2-20 were screened for their leishmanicidal effects using amphotericin B (IC(50) = 0.24 microg L(-1)) and pentamidine (IC(50) = 2.56 microg mL(-1)) as standards and a structure-activity relationship (SAR) study was established. The compounds 2 (IC(50) = 48 microg mL(-1)), 3 (IC(50) = 30.0 microg mL(-1)), 10 (IC(50) = 41.0 microg mL(-1)), 15 (IC(50) = 33.0 microg mL(-1)), 16 (IC(50) = 35.0 microg mL(-1)) and 20 (IC(50) = 47.0 microg mL(-1)) showed weak leishmanicidal activities. PMID- 19296394 TI - A convenient synthesis of p-substituted 1-arylsulfonyl-pyrrolidin-2-ones. AB - A convenient and novel method of cyclisation of 4-(4-substituted phenylsulfonamido)-butanoic acids to their corresponding p-substituted 1 arylsulfonyl-pyrrolidin-2-ones was achieved by using polyphosphate ester (PPE). The reaction times were considerably reduced, with an increase in yields, when PPE was used in combination with a catalytic amount of pyridine. PMID- 19296395 TI - New acylated flavone and cyanogenic glycosides from Linum grandiflorum. AB - The first investigation of Linum grandiflorum resulted in the isolation of one new acylated flavone O-diglycoside known as luteolin 7-O-alpha-D-(6'''-E feruloyl)glucopyranosyl (1 --> 2)-beta-D-glucopyranoside, and one new cyanogenic glycoside known as 2-[(3'-isopropoxy-O-beta-D-glucopyranosyl)oxy]-2 methylbutanenitrile, together with four known flavonoid glycosides, three known cyanogenic glycosides and one alkyl glycoside. The new compounds were structurally elucidated via the extensive 1D, 2D NMR and DIFNOE together with ESI TOF-CID-MS/MS and HR-MALDI/MS. PMID- 19296396 TI - New furanocoumarins detected from grapefruit juice retentate. AB - Grapefruit (Citrus paradisi Macf.) furanocoumarins and related compounds have been shown to interact with the enterocyte cytochrome P450, CYP3A4, and as a result they affect the bioavailibility of certain drugs. Only a few grapefruit furanocoumarins have been identified so far. In this study, grapefruit juice retentate, rich in furanocoumarins, was extracted and then separated by flash chromatography for the examination of new compounds. Finally, nine new furanocoumarins were detected in different fractions according to their UV spectra and mass spectrometric properties by LC-MS (liquid chromatography mass spectrometry) and tentatively designated as FC 338, FC 420, FC 524, FC 530, FC 540, FC 546, FC 552, FC 570 and FC 614. PMID- 19296397 TI - Impairments in prospective and retrospective memory following stroke. AB - Prospective memory (PM) is the ability to carry out a planned intention at a future time. We studied PM deficits in a group of community-dwelling stroke survivors compared with normal controls. Twelve stroke patients and 12 matched controls performed a series of tests assessing executive function, prospective (PM) and retrospective memory (RM). Patients performed less well than controls on laboratory measures of PM and associative RM; they also showed deficits on standard tests of RM and executive control. The groups did not differ on more structured clinical measures of executive function, RM, PM or self-rated PM and RM. The results are discussed in terms of an impairment in the ability to 'self initiate' effortful cognitive processes. PMID- 19296398 TI - Smoking is associated with steatosis and severe fibrosis in chronic hepatitis C but not B. AB - OBJECTIVE: The results of retrospective studies suggest an association between smoking, insulin resistance, steatosis and fibrosis in patients with chronic hepatitis C (CHC); no data are available for chronic hepatitis B (CHB). The purpose of this study was to evaluate the relationship, if any, of such factors on liver fibrosis in a cohort of patients with CHB and CHC. MATERIAL AND METHODS: The study prospectively included 271 consecutive patients with CHB (n=95) or CHC (n=176) who had undergone liver biopsies. Each patient completed a questionnaire on smoking habits; anthropometric measurements and laboratory examinations were carried out and histological lesions were recorded. RESULTS: In CHC patients, severe fibrosis was independently associated with a higher body mass index (BMI) (OR: 1.180, 95% CI: 1.028-1.354; p=0.019), heavy smoking (OR: 3.923, 95% CI: 1.356-11.348; p=0.012), higher alanine aminotransferase (ALAT) levels (OR: 1.010, 95% CI: 1.003-1.017; p=0.005) and alkaline phosphatase (ALP) levels (OR: 1.016, 95% CI: 1.001-1.030; p=0.03) and presence of necroinflammation (OR: 11.165, 95% CI: 1.286-96.970; p=0.029). Moreover, steatosis was independently associated with high gamma-glutamyl transpeptidase (GGT) values, heavy smoking and presence of necroinflammation. In CHB patients, no association between smoking habits and fibrosis or steatosis was noted. CONCLUSIONS: Heavy smoking is associated with severe fibrosis in CHC but not CHB. Heavy smoking is also significantly associated with steatosis in CHC and this could be the link between smoking and fibrosis progression. PMID- 19296399 TI - Chronic asymptomatic pancreatic hyperenzymemia is a benign condition in only half of the cases: a prospective study. AB - OBJECTIVE: Prospectively to evaluate patients with chronic asymptomatic pancreatic hyperenzymemia in order to identify possible pancreatic and non pancreatic diseases. MATERIAL AND METHODS: Seventy-five asymptomatic subjects with long-standing pancreatic hyperenzymemia (45 M, 30 F; mean age+/-SD 51.5+/ 16.0 years, range 19-78 years, mean duration+/-SD of pancreatic hyperenzymemia 14.7+/-7.0 months, range 7-34 months) and normal ultrasonographic evaluation were included in this study. The subjects enrolled were carefully interviewed and prospectively evaluated. All patients underwent blood screening. An additional abdominal ultrasound was also carried out and, if considered necessary, other imaging and endoscopic evaluation procedures were used. RESULTS: The follow-up of the patients after enrollment in the study was 3.3+/-1.8 years (mean+/-SD). In 38 patients (50.7%), pancreatic or extrapancreatic disease was diagnosed: 20 patients had chronic pancreatitis, 1 had autoimmune chronic pancreatitis, 1 had a benign cyst of the pancreas, 2 had serous cystadenomas, 5 had an intraductal papillary mucinous tumor of the pancreas, 3 had a ductal pancreatic adenocarcinoma, 4 patients had chronic viral hepatitis, and 2 had Crohn's disease. In 37 subjects (49.3%), no pancreatic or extrapancreatic diseases were found (3 subjects had macroamylasemia, 3 had familial hyperenzymemia, 31 had chronic non-pathological pancreatic hyperenzymemia). CONCLUSIONS: Subjects having an increase of either amylase or lipase serum levels should undergo a thorough diagnostic work-up prior to establishing the existence of chronic non pathological pancreatic hyperenzymemia. PMID- 19296400 TI - Is length of experience an appropriate criterion to identify level of expertise? AB - Clinical experience plays an important role in the development of expertise, particularly when coupled with reflection on practice. There is debate, however, regarding the amount of clinical experience that is required to become an expert. Various lengths of practice have been suggested as suitable for determining expertise, ranging from five years to 15 years. This study aimed to investigate the association between length of experience and therapists' level of expertise in the field of cerebral palsy with upper limb hypertonicity using an empirical procedure named Cochrane-Weiss-Shanteau (CWS). The methodology involved re analysis of quantitative data collected in two previous studies. In Study 1, 18 experienced occupational therapists made hypothetical clinical decisions related to 110 case vignettes, while in Study 2, 29 therapists considered 60 case vignettes drawn randomly from those used in Study 1. A CWS index was calculated for each participant's case decisions. Then, in each study, Spearman's rho was calculated to identify the correlations between the duration of experience and level of expertise. There was no significant association between these two variables in both studies. These analyses corroborated previous findings of no association between length of experience and judgemental performance. Therefore, length of experience may not be an appropriate criterion for determining level of expertise in relation to cerebral palsy practice. PMID- 19296401 TI - The extent and pattern of organ damage in small vessel vasculitis measured by the Vasculitis Damage Index (VDI). AB - OBJECTIVES: To assess the extent and pattern of irreversible organ damage in patients with Wegener's granulomatosis (WG), microscopic polyangiitis (MPA), polyarteritis nodosa (PAN), and Churg-Strauss syndrome (CSS) by a cross-sectional point prevalence study within a defined geographical area. METHODS: The Vasculitis Damage Index (VDI) was recorded for 86 prevalent cases, classified as 46 patients with WG, 27 with MPA, nine with PAN, and four with CSS from a defined population in southern Sweden, with a median age of 64.8 years and a median disease duration of 9 years. The VDI was determined for all patients at the day of point prevalence (pp), 1 January 2003. RESULTS: The median VDI score was 3 [interquartile range (IQR) 2-5] for all patients: 3 (2-4) for WG, 3 (1.5-4.5) for MPA, 5 (2-6) for PAN, and 1.5 (0.75-2.75) for CSS. Only 9% of patients had not been assigned a single item of damage. The most common damage was cardiovascular, followed by renal, neuropsychiatric, ear nose and throat (ENT), and musculoskeletal. Major vascular and treatment-related damage was associated with advanced age whereas ENT damage was more prevalent in younger patients. There was an almost complete separation between ENT damage and cardiac and renal damage with only two out of the 22 patients assigned ENT damage having experienced renal damage; none had been assigned cardiac damage. Patients with cardiac damage had significantly higher damage rates. CONCLUSIONS: Damage remains an important problem for patients with systemic vasculitis despite effective remission inducing drugs. Only a small fraction of patients are unmarked by their disease. PMID- 19296402 TI - A longitudinal follow-up of hand involvement and activities of daily living in early systemic sclerosis. AB - OBJECTIVE: To investigate the development of hand involvement and activities of daily life (ADL) in early systemic sclerosis (SSc), and to examine the usefulness of the Hand Mobility in Scleroderma (HAMIS) test in a longitudinal study. METHODS: Forty-three patients with disease duration no longer than 3 years and at least 4 years of follow-up formed the study group. Based on skin involvement, 31 patients were classified as limited cutaneous SSc (lcSSc) and 12 as diffuse cutaneous SSc (dcSSc). The patients were assessed regarding hand function [mobility (HAMIS), grip force (Grippit), and self-assessed hand function visual analogue scale (VAS)], vascular involvement [Raynaud's phenomenon (RP)], skin involvement [modified Rodnan skin score (mRss)], and ADL capacity [the scleroderma Functional Score (FS)]. RESULTS: Hand mobility and ADL capacity were in general good and did not change significantly. However, at the individual level 72% of the patients showed a change in HAMIS score. Grip force and perceived hand function were moderately impaired at baseline and during the follow-up. Skin involvement in the hand/arm improved significantly (p<0.001). During the observation period, the hand/arm mRss and HAMIS score changed in parallel (r(s) = 0.58, p<0.001). ADL capacity correlated significantly with grip force, self-assessed hand function, and RP at baseline, and also with HAMIS at follow-up. CONCLUSIONS: In this study hand mobility and ADL capacity were maintained during the first years of SSc. HAMIS was found to be a feasible test for longitudinal assessment of hand mobility in SSc. PMID- 19296403 TI - Annual incidence and standardized incidence ratio of cerebrovascular accidents in patients with systemic lupus erythematosus. AB - OBJECTIVES: To study the annual incidence and standardized incidence ratio (SIR) of cerebrovascular accident (CVA) in patients with systemic lupus erythematosus (SLE). SUBJECTS AND METHODS: The annual incidence of CVA from 1999 to 2007 in a longitudinal cohort of SLE patients was calculated each year and compared with that of the regional population within the same study period. Age-specific SIRs and outcome of CVA in SLE patients were also studied. RESULTS: In 2007, there were 490 SLE patients in our cohort. The mean annual incidence of CVA between 1999 and 2007 was 6.45/1000 patients and no obvious trend over time was observed. Of the 20 CVAs in patients with SLE, 18 (90%) were ischaemic stroke whereas two (10%) were haemorrhagic stroke. The mean SIR of all types of CVA in SLE patients was 2.02 [95% confidence interval (CI) 1.30-3.81; p = 0.002]. The SIR of ischaemic stroke decreased with age and the stroke incidence was no longer significantly higher than that of the population in patients aged >or= 60 years. Haemorrhagic stroke occurred mainly in younger SLE patients. The duration of hospitalization and the mortality rate for CVA was non-significantly higher in SLE than in non-SLE patients. CONCLUSIONS: The incidence of CVA in SLE remained constant over the 8 years between 1999 and 2007. Younger SLE patients are at substantially increased risk of CVA compared to age-matched population. The duration of hospitalization and the mortality rate for CVA are similar in SLE and non-SLE patients. PMID- 19296404 TI - Prevalence of antibodies against Chlamydia trachomatis and incidence of C. trachomatis-induced reactive arthritis in an early arthritis series in Finland in 2000. AB - OBJECTIVE: To study the prevalence of different serotypes of Chlamydia trachomatis antibodies and the incidence of C. trachomatis-induced reactive arthritis (ReA) among patients with early arthritis in a defined population. METHODS: Serum samples were collected from a cohort of 122 adult patients in the age group 18-65 years included in the Kuopio 2000 Arthritis Survey. Antibodies against C. trachomatis serotypes C, E, and G were studied using enzyme immunoassay (EIA) tests among patients and in a control cohort of 78 adults without any joint symptoms. The incidence assessment for Chlamydia-induced ReA was based on a ligase chain reaction (LCR) test in urine and clinical symptoms and signs appropriate for ReA. RESULTS: Of 122 patients, with the baseline diagnosis of rheumatoid arthritis (RA) in 11, spondyloarthropathy (SpA) in 28, and undifferentiated arthritis (UA) in 83 cases, 42 (34%) showed immunoglobulin (Ig)G or IgA antibodies against at least one serotype C, E, or G. Among the patients with UA the prevalence was significantly increased compared with the controls (p = 0.010). C. trachomatis-induced ReA arthritis was diagnosed in only three patients with the LCR test. On this basis the incidence of C. trachomatis induced arthritis was 5.4/100 000 [95% confidence interval (CI) 1.1-15.7] in the age group 18-65 years. CONCLUSION: Antibodies against C. trachomatis were most common in patients with UA reflecting the fact that cases with chlamydia-induced ReA are included in this subgroup. PMID- 19296405 TI - Proteases from salmon stimulate IL-8 in airway epithelial cells. AB - In this study the ability of salmon tissue extracts to stimulate interleukin 8 (IL-8) production in airway epithelial cells (A549) was investigated; in particular, the role of serine protease enzymes and endotoxin was examined with respect to IL-8-stimulating ability. A549 cells were stimulated by various concentrations of fish tissue extracts for 6 h. Parallel samples were incubated with a protease inhibitor cocktail, a serine protease inhibitor, or an endotoxin inhibitor. The amount of secreted IL-8 in the supernatant was determined by an enzyme-linked immunosorbent assay. A549 cells showed a concentration-dependent increase in IL-8 secretion after stimulation with extracts of salmon tissues. The IL-8-stimulating effect was inhibited by serine protease inhibitors but not by endotoxin inhibitors. PMID- 19296406 TI - Trace elements in Pandanus (Pandanus tectorius) from a manganese-enriched wetland in Southern Guam: a possible Lytico-Bodig connection? AB - Extremely high levels of manganese (Mn) were encountered in foliar tissue of the monocot tree Pandanus tectorius from southern Guam with values exceeding 10,000 microg/g dry weight in some wetland representatives. Historically, dried Pandanus leaves were used extensively as a source of domestic fiber in the local Chamorro culture. A possible link between the use of this plant and a neurodegenerative disease complex that once plagued the island and is symptomatically similar to the occupational disease "manganism" poses an intriguing question that merits further investigation. PMID- 19296407 TI - Molecular cloning and oxidative modification of human lens ALDH1A1: implication in impaired detoxification of lipid aldehydes. AB - Earlier studies showed that human lens ALDH1A1 plays a critical role in protection against oxidative stress-induced cytotoxicity in human lens epithelial cells (HLEC), and opacification of rat and mouse lens. The complete coding sequence of ALDH1A1 was cloned from human lens cDNA library by using PCR methods and expressed it in Escherichia coli. The cloned human lens ALDH1A1 cDNA encodes a 501-amino-acid protein (molecular mass = 54.8 kD) that is 100% identical to human liver ALDH1A1 and shares significant identity with the same isozyme from other tissues and species. The purified recombinant human lens ALDH1A1 exhibited optimal catalytic activity at pH 8 and preferred NAD(+) as cofactor and specifically catalyzed the oxidation of toxic lipid aldehydes such as 4 hydroxynonenal (HNE; K(m) = 4.8 microM) and malonaldehyde (K(m) MDA = 3.5 microM). Citral, disulfiram, and cyanamide were found to inhibit human lens ALDH1A1 at IC50 values of 55, 101, and 22610 microM, respectively, whereas diethylstilbestrol (DES) was found to be an activator (EC(50), 1.3 microM). Further, modification of recombinant human lens ALDH1A1 with nitric oxide donors such as S-nitroso-N-acetylpenicillamine (SNAP) and S-nitrosoglutathione (GSNO) significantly inhibited the enzyme activity. It therefore appears that activation of ALDH1A1, which efficiently catalyzes the detoxification of lipid-derived toxic aldehydes, and/or prevention of its oxidative modification may be novel therapeutic interventions against oxidative stress-induced lens pathologies. PMID- 19296408 TI - The concentration of no toxicologic concern (CoNTC) and airborne mycotoxins. AB - The threshold of toxicologic concern (TTC) concept was developed as a method to identify a chemical intake level that is predicted to be without adverse human health effects assuming daily intake over the course of a 70-yr life span. The TTC values are based on known structure-activity relationships and do not require chemical-specific toxicity data. This allows safety assessment (or prioritization for testing) of chemicals with known molecular structure but little or no toxicity data. Recently, the TTC concept was extended to inhaled substances by converting a TTC expressed in micrograms per person per day to an airborne concentration (ng/m(3)), making allowance for intake by routes in addition to inhalation and implicitly assuming 100% bioavailability of inhaled toxicants. The resulting concentration of no toxicologic concern (CoNTC), 30 ng/m(3), represents a generic airborne concentration that is expected to pose no hazard to humans exposed continuously throughout a 70-yr lifetime. Published data on the levels of mycotoxins in agricultural dusts or in fungal spores, along with measured levels of airborne mycotoxins, spores, or dust in various environments, were used to identify conditions under which mycotoxin exposures might reach the CoNTC. Data demonstrate that airborne concentrations of dusts and mold spores sometimes encountered in agricultural environments have the potential to produce mycotoxin concentrations greater than the CoNTC. On the other hand, these data suggest that common exposures to mycotoxins from airborne molds in daily life, including in the built indoor environment, are below the concentration of no toxicologic concern. PMID- 19296409 TI - Catechol O-methyltransferase (COMT) VAL158MET functional polymorphism, dental mercury exposure, and self-reported symptoms and mood. AB - Associations were evaluated between a functional single nucleotide polymorphism (Val158Met) in the gene encoding the catecholamine catabolic enzyme catechol O methyltransferase (COMT), dental mercury exposure, and self-reported symptoms and mood among 183 male dentists and 213 female dental assistants. Self-reported symptoms, mood, and detailed work histories were obtained by computerized questionnaire. Spot urine samples were collected and analyzed for mercury concentrations to evaluate recent exposures, whereas a chronic mercury exposure index for all subjects was created from the work histories. COMT polymorphism status was determined using a polymerase chain reaction (PCR)-based assay. Scores for current, recent, and chronic self-reported symptom groups and six self reported mood factors were evaluated with respect to recent and chronic mercury exposure and COMT polymorphism status. Multiple regression analysis controlled for age, socioeconomic status, tobacco and alcohol use, self-reported health problems, and medications. Separate evaluations were conducted for dentists and dental assistants. No consistent patterns of association between either urinary mercury concentration or the chronic index of mercury exposure and any category of symptoms were observed. However, consistent and significant associations were found between increased symptoms and the COMT polymorphism involving the double allelic substitution (full mutation) compared to subjects with no substitutions. Associations with mood were limited to polymorphism status among female dental assistants, and were observed for four of six mood factors and overall mood score. These findings extend evidence of genetic factors potentially affecting human susceptibility to the toxic effects of mercury and other environmental chemicals. PMID- 19296410 TI - Spatial variability of volatile organic compound concentrations in Sarnia, Ontario, Canada. AB - As part of a community health study, an air quality investigation was carried out in Sarnia, Ontario, Canada, which is located within a government-designated "Area of Concern". The Sarnia area, also known as Chemical Valley, has been known to have an abundance of air quality problems that are frequently attributed to the large number of chemical companies located in the area, which are among the largest industrial sources of criteria air pollutants in Canada. Ambient volatile organic compounds (VOC) were monitored during a 2-wk period in October 2005 at 37 sites across the city of Sarnia, using 3M organic vapor monitors. The spatial variability was investigated using ranking and mapping techniques to identify spatial patterns of the VOC across the city. The results show a large spatial variability of most of the VOC investigated. Furthermore, most of the VOC were highly correlated, suggesting common sources in Sarnia. The results also indicate that the industrial and chemical facilities strongly influence the concentrations of VOC in Sarnia, with higher outdoor VOC levels found closer to industrial facilities. The measured VOC and those obtained from a long-term government monitoring station were compared. It was determined that the exposure gradients observed during the study period are relatively representative of the long-term pattern. The results of this study will be used for the community health study. PMID- 19296411 TI - Community-acquired pneumonia. PMID- 19296412 TI - Global and local epidemiology of community-acquired pneumonia: the experience of the CAPNETZ Network. AB - Community-acquired pneumonia (CAP) is a major cause of morbidity and mortality worldwide. Young children and the elderly are disproportionately affected by CAP. Lower respiratory tract infections (LRTIs), including CAP, were ranked third in a list of the 30 leading causes of death worldwide in 1990. Mortality rates are low (< 2%) in CAP patients treated as outpatients, but are higher (5 to 20%) among patients hospitalized for CAP, and are highest (up to 50%) in patients admitted to the intensive care unit. Several risk factors are known to be associated with increases in mortality, the most important of which are age > 65 years, male gender, and comorbidities such as chronic heart failure, advanced chronic obstructive pulmonary disease, neurological diseases, and liver cirrhosis. Patients living in nursing homes may have a special risk for multiresistant bacterial infection. The incidence of CAP varies worldwide by country, age, and gender. Further, data about epidemiology, etiology, morbidity, mortality, and economical burden of diseases differ between countries. In this review, we present recent data regarding the incidence, etiology, and rate of antibiotic resistance among CAP patients from the German Network for Community Acquired Pneumonia (CAPNETZ) registry and review data from several European countries. PMID- 19296413 TI - The European vision of community-acquired pneumonia. AB - Community-acquired pneumonia is common throughout the 53 European countries. Despite differences in population structure and habits there are many similarities between countries, including overall frequency and distribution of major causative pathogens. There are, however, also differences between countries, particularly in the frequency of bacterial antibiotic resistance and to a lesser extent how the condition is managed. Having said this there are many gaps in our knowledge with little or no data available from some countries, especially those in the eastern Europe. It is hoped that future years will see these gaps filled through continuing research. PMID- 19296414 TI - Adjunctive therapy in community-acquired pneumonia. AB - Despite potent antibiotics, community-acquired pneumonia (CAP) remains the most common cause of death from infection and the seventh overall leading cause of death in the United States. For this reason, interest has been redirected into non-antibiotic therapeutic measures. Despite theoretical benefits, the existing literature does not suggest a clear benefit for corticosteroid treatment, but large prospective randomized trials are needed. Nonsteroidal antiinflammatory drugs may benefit oxygenation but have no documented effect on mortality. Activation of the coagulation system appears to be a major pathophysiological event in severe pneumonia, possibly even more so than for sepsis in general. The CAP subgroup in phase III sepsis trials of both drotrecogin alfa (activated) and tifacogin (recombinant tissue factor pathway inhibitor) demonstrated the greatest benefit. The immunomodulatory effects of macrolide antibiotics may play a significant role in management of severe CAP. Exogenous surfactant replacement is being explored as adjunctive therapy for acute lung injury due to CAP. Statin use before CAP diagnosis is associated with improved outcome but requires further research to determine if initiation at the time of diagnosis will affect outcome. Other therapies have theoretical benefit but are not yet in the stage of clinical trials. PMID- 19296415 TI - Early and late treatment failure in community-acquired pneumonia. AB - Treatment failure is a matter of great concern in the management of community acquired pneumonia (CAP). Defined generally as lack of response or clinical deterioration, failure is considered early when it occurs within the first 72 hours and late when it occurs after 72 hours. The reported incidence of treatment failure among hospitalized patients with CAP ranges from 2.4 to 31% for early failure and from 3.9 to 11% for late failure. Most cases of early failure occur because of inadequate host-pathogen responses. Factors associated with treatment failure include high-risk pneumonia, liver disease, multilobar infiltrates, Legionella pneumonia, gram-negative pneumonia, pleural effusion, cavitation, leucopenia, and discordant antimicrobial therapy. Conversely, influenza vaccination, initial treatment with fluoroquinolones, and chronic obstructive pulmonary disease have been linked with a lower risk of failure. Treatment failure is associated with high morbidity and mortality rates. Its detection and management require careful clinical assessment. Certain serum biological markers may be helpful to identify patients with a higher risk of deterioration and poor prognosis. Because inadequate host-pathogen responses are responsible for a significant number of failures, strategies aimed at modulating the inflammatory response should be investigated. Discordant therapy can be prevented by rational application of the current antibiotic guidelines. PMID- 19296416 TI - New antimicrobial molecules and new antibiotic strategies. AB - Drug options for treatment of infections are increasingly limited. The pharmaceutical industry has found it difficult to discover new antimicrobial agents, and only two novel classes of antibiotics, the oxazolidinones and the cyclic lipopeptides, have entered the market since the late 1960s. Few new agents have reached the market in the last decade with potential interest for community acquired pneumonia (CAP) treatment, including linezolid (the first oxazolidinone in clinical use), new fluoroquinolones, cefditoren, ertapenem, and telithromycin. Agents currently in clinical development include other novel quinolones and ketolides, broad-spectrum cephalosporin derivatives, faropenem, several glycopeptides, and iclaprim. Other molecules are considered to be promising candidates for the future. In addition to the foregoing agents, alternative treatment approaches have also been introduced into clinical practice, which include the administration of the appropriate antimicrobials in a timely manner and the consideration of the pharmacokinetic-pharmacodynamic properties of the agent(s). PMID- 19296417 TI - Impact of guidelines on outcome: the evidence. AB - Pneumonia continues to be the main cause of death due to infection in the world, and it produces a high consumption of healthcare resources. The guidelines established by the scientific societies improve the care of patients with pneumonia. One way of evaluating the effect of the guidelines is to analyze their impact on the prognosis of the infection. To evaluate this effect, cohort studies have been performed using before-after, observational, cost-effectiveness, and, to a lesser degree, randomized designs. The most recent studies show that the implementation of the guidelines is accompanied by an increase in the process of care percentage and a lower inpatient hospital mortality rate- including the first 48 hours and after 30 days. These findings are consistent across various studies, and they have been confirmed in patients admitted to the intensive care unit. Clinical stability is also reached earlier in patients hospitalized for community-acquired pneumonia (CAP) when the antibiotic treatment is begun early and complies with the recommendations. Finally, the choice of antibiotics that adhere to the guidelines is cost-effective in CAP requiring hospitalization, which is responsible for 80% of the total cost of this disease. PMID- 19296418 TI - Community-acquired pneumonia: the U.S. perspective. AB - Community-acquired pneumonia (CAP) is a common and serious problem in the United States, and the sixth leading cause of death in those over age 65. Not only has short-term mortality been evaluated, but 1-year mortality may be as high as 40% in Medicare patients who have been admitted to the hospital with CAP. In the United States, guidelines for CAP management have been available since 1993, with the most recent version published in 2007 as a joint effort of the Infectious Diseases Society of America and the American Thoracic Society. The current U.S. guidelines take into consideration unique bacteriologic patterns in the United States, particularly focusing on the role of drug-resistant pneumococcus, atypical pathogens, and methicillin-resistant Staphylococcus aureus, which explains why U.S. recommendations for therapy differ from those in Europe and the United Kingdom. Notable differences in the U.S. approach to CAP compared with elsewhere include not only a unique set of bacteriologic considerations and therapy recommendations that follow these concerns but also a different approach to assessing severity of illness and recommended diagnostic testing, as well as the inclusion of performance measures to optimize disease management. Compared with European and British guidelines, the U.S. therapy of CAP has a greater emphasis on the role of atypical pathogens, a more defined role for fluoroquinolones as first-line therapy, less reliance on oral therapy for hospitalized patients, and less regard for the value of certain beta-lactam agents. PMID- 19296419 TI - Streptococcus pneumoniae: epidemiology, risk factors, and strategies for prevention. AB - Streptococcus pneumoniae is the most common cause of community-acquired pneumonia, meningitis, and bacteremia in children and adults. Invasive pneumococcal disease (IPD) primarily affects young children, older adults (> 65 years of age), and individuals with comorbidities or impaired immune systems. Case fatality rates range from 10 to 30% in adults with IPD but are much lower (< 3%) in children. In this article, we describe the epidemiology of IPD, risk factors, and the influence of host- and organism-specific factors on outcomes. Most cases of IPD are caused by a limited number of serotypes that vary in infectivity and virulence. Vaccinating adults and high-risk patients with the pneumococcal polysaccharide vaccine reduces the incidence of IPD in populations at risk but does not affect nasopharyngeal colonization and has had limited benefit in the population at large. Use of the heptavalent pneumococcal conjugate vaccine (PCV7) in children in the United States since 2000 has resulted in a substantial decline of IPD in both children and adults (by herd immunity), but has facilitated the emergence of serotypes not encompassed in the PCV7 vaccine. Recent reports of "replacement" serotypes that have heightened virulence are worrisome. In this chapter, we discuss the role of vaccines (both polysaccharide and conjugate) and other preventive strategies to limit this important and potentially lethal disease. PMID- 19296420 TI - Streptococcus pneumoniae: does antimicrobial resistance matter? AB - Over the past 3 decades, antimicrobial resistance among Streptococcus pneumoniae, the most common cause of community-acquired pneumonia (CAP), has escalated dramatically worldwide. In the late 1970s, strains of pneumococci displaying resistance to penicillin were described in South Africa and Spain. By the early 1990s, penicillin-resistant clones of S. pneumoniae spread rapidly across Europe and globally. Additionally, resistance to macrolides and other antibiotic classes escalated in tandem with penicillin resistance. Six international clones (serotypes 6A, 6B, 9V, 14, 19F, 23F) were responsible for most of these resistant isolates. Currently, 15 to 30% of S. pneumoniae worldwide are multidrug-resistant (MDR) (i.e., resistant to > or = 3 classes of antibiotics). Despite the dramatic escalation in the rate of antimicrobial resistance among pneumococci worldwide, the clinical impact of antimicrobial resistance is difficult to define. Treatment failures due to antibiotic-resistant pneumococci have been reported with meningitis, otitis media, and lower respiratory tract infections, but the relation between drug resistance and treatment failures has not been convincingly established. Clinical failures often reflect factors independent of antimicrobial susceptibility of the infecting organisms. Host factors (e.g., extremes of age; underlying immunosuppressive or debilitating disease; comorbidities), or factors that affect intrinsic virulence of the organisms (e.g., capsular subtype) strongly influence prognosis. Mortality rates are higher in the presence of: multilobar involvement, renal insufficiency, need for intensive care unit (ICU) care, hypoxemia, severe derangement in physiological parameters, and comorbidities. Given these confounding factors, dissecting out the impact of antimicrobial resistance on clinical outcomes is difficult, if not impossible. Prospective, randomized trials designed to assess the clinical significance of antimicrobial resistance among pneumococci are lacking, and for logistical reasons, will never be done. Does in vitro resistance translate into clinical failures? Should changing resistance patterns modify our choice of therapy for CAP or for suspected pneumococcal pneumonia? In this review, we discuss several facets, including mechanisms of antimicrobial resistance among specific antibiotic classes, epidemiology and spread of antimicrobial resistance determinants regionally and worldwide, risk factors for acquisition and dissemination of resistance, the impact of key international clones displaying multidrug resistance, the clinical impact of antimicrobial resistance, and strategies to limit or curtail antimicrobial resistance among this key respiratory tract pathogen. PMID- 19296421 TI - Current perspective of the HCAP problem: is it CAP or is it HAP? AB - The number of individuals receiving health care outside the hospital setting, including home wound care or infusion therapy, dialysis, nursing homes, and similar settings is constantly increasing. One of the most frequent causes of hospitalization and mortality in these patients is pneumonia. Hence a new class of pneumonia has been identified: healthcare-associated pneumonia (HCAP). The last American Thoracic Society/Infectious Disease Society of America (ATS/IDSA) guidelines define specific criteria to identify HCAP; however, the clinical practice suggests that the presence of indwelling devices (permanent catheters, etc.) may also be considered an additional criterion. Different studies have shown that, in comparison with community-acquired pneumonia (CAP) patients, HCAP patients are significantly older, have a higher number of comorbidities (cerebrovascular diseases, congestive heart failure, dementia, and diabetes mellitus) and show worse functional status before admission. It has also been observed that HCAP differs from CAP in terms of clinical presentation, risk factors, etiology, prognostics, and, likely, therapeutic approach. The clinical presentation of HCAP is often unusual because it is frequently conditioned by advanced age, multiple chronic comorbidities, and neurological disorders. Classic respiratory symptoms of pneumonia are often mild in HCAP, whereas extrapulmonary manifestations, including mental confusion and gastrointestinal disorders, are frequent. HCAP patients, commonly present a worse clinical presentation (hypoxemia, altered consciousness, Fine score, multilobar infiltrates, etc.) than CAP, and a mortality rate close to that of hospital-acquired pneumonia. Many studies have attributed these findings to a nosocomial etiology [methicillin resistant Staphylococcus aureus (MRSA) , Pseudomonas aeruginosa, etc.] with a high frequency of multidrug-resistant infections (MRIs), even though this remains controversial. Further investigation on microbial composition and MRI risk factors of HCAP is fundamental because no definitive therapeutic indications are currently available. PMID- 19296422 TI - [Editorial comment on: Arsov C. et al.: prostate cancer centres/prostate centres- certification by DKG and DVPZ]. PMID- 19296423 TI - [Prostate cancer centres / prostate centres--certified by DKG or DVPZ]. AB - In Germany, ambitious efforts for the implementation of well-defined quality standards in the diagnosis of and therapy for prostate cancer have resulted in the creation of certified centres over the past years. This development is in analogy to that of certified breast and intestinal centres. Concerning the prostate, there are two different models for the certification of centres. The model of the Dachverband der Prostatazentren e. V. (DVPZ) includes the most common kinds of dis-eases of the prostate, namely BPH, prostate cancer and prostatitis. On the other hand prostate cancer centres are certified by the Deutsche Krebsgesellschaft (DKG). Among German urologists there is a controversial discussion concerning this process. The present paper gives a review of the two models and discusses the chances as well as the risks which come along with certified centres. PMID- 19296425 TI - [Inguinal lymphadenectomy in penis carcinoma]. PMID- 19296424 TI - [Anti-inflammatory action of a hyaluronic acid-chondroitin sulfate preparation in an in vitro bladder model]. AB - PURPOSE: Interstitial cystitis and BPS (bladder pain syndrome) are chronic inflammatory diseases of the bladder. They are as yet imperfectly understood diseases, possibly originating from damage to the glycosaminoglycan layer of the bladder epithelium . Hyaluronic acid-containing preparations are currently utilised for palliation of the symptoms and protection of the bladder epithelium . The aim of the work described here was the evaluation of one of these preparations containing chondroitin sulfate together with hyaluronic acid. MATERIAL AND METHODS: The preparation was evaluated for its anti-inflammatory potential as well as regarding the tolerance by the bladder epithelium. The urothelial cell line T24 was employed as an in vitro model of the human bladder because of its ability to react to adequate stimuli with the release of interleukin 6. To this end the cells were treated with hyaluronic acid and chondroitin sulfate. Subsequently, TNF-alpha was applied to induce inflammation. The severity of inflammation was measured on the basis of the IL-6 released by the cells in comparison to untreated control cultures. RESULTS: A reduction of TNF-alpha-induced IL-6 release after treatment with hyaluronic acid and chondroitin sulfate was observed, indicating the anti-inflammatory action of the preparation. As shown by the large number of living cells after treatment the test preparation did not affect cell viability even in high concentrations. These data suggest a good tolerance of the product by the patients. CONCLUSION: The administration of the preparation in patients suffering from interstitial cystitis or BPS appears promising. In additional, the presented work demonstrates the feasibility of the cell culture model for the screening of new therapeutic approaches. PMID- 19296426 TI - [Is olfactory function impaired in moderate height?]. AB - BACKGROUND: The human sense of smell seems to be influenced by the surrounding barometric pressure. These factors appear to be especially important during flights, for example, in order to recognize the smell of fire etc. Thus, questions are whether pilots or passengers exhibit an impaired smell sensitivity when tested at moderate heights, or, whether changes in humidity would affect the sense of smell. MATERIAL AND METHOD: Using climate chambers, odor discrimination and butanol odor thresholds were tested in 77 healthy normosmic volunteers (5 female, 72 male; aged 25+/-8 years from 18 up to 53 years) under hypobaric (2 700+/-20 m, 20 degrees C+/-1 K, rh=50+/-5%) and hyperbaric, (10+/-0.5 m (2 bar)) and different humidity conditions (30 vs. 80%, 20 degrees C+/-1 K, normobaric). During all conditions cognitive performance was tested. RESULTS: Among other effects, olfactory sensitivity was impaired at threshold, but not suprathreshold level, in a hypobaric compared to a hyperbaric milieu, and thresholds were lower in humid, compared to relatively dry conditions. CONCLUSIONS: In conclusion, environmental conditions modulate the sense of smell, and may, consecutively, influence results from olfactory tests. During flight hypobaric conditions, mild hypoxia and dry air may cause impaired sensitivity of smell. PMID- 19296427 TI - The diterpenoid ent-16-kauren-19-oic acid acts as an uncoupler of the bacterial respiratory chain. AB - ent-16-Kauren-19-oic acid is a bacteriolytic diterpenoid present in the resin of the medicinal plant Pseudognaphalium vira vira. The compound and its methyl ester showed strong activity against gram-positive bacteria, whereas the derivative 3beta-hydroxy-ent-kauren-19-oic acid was inactive against all assayed bacteria at the maximal concentration used (250 microg/mL). The bacteriolytic effect of ent 16-kauren-19-oic acid (5 microg/mL) was confirmed with cultures of Bacillus cereus and Staphylococcus aureus, whereas the methyl derivative (12 microg/mL) showed only a bacteriostatic effect. Both compounds stimulated oxygen consumption and proton conduction of whole cells, as reflected by an abrupt increase in the extracellular pH. These results indicate that ent-16-kauren-19-oic acid acts as a respiratory chain uncoupler, and that this function is strongly affected by minor structural substitutions, suggesting a tight activity-structure relationship. The ultimate effect of the uncoupling mechanism demonstrated by ent-16-kauren-19-oic acid is bacterial lysis. The disruption of the bacterial membrane integrity caused by the diterpenoid compound was determined using SYTOX Green stain and visualized by fluorescence microscopy. PMID- 19296428 TI - Qianliguang (Senecio scandens) safety dilemma: dose is the key? AB - Qianliguang ( SENECIO SCANDENS) is a common Chinese medicinal herb. Qianliguang containing herbal proprietary products are registered as over-the-counter remedies in China and exported to Western countries. The safety of using Qianliguang and its products has raised general concerns because of a potential risk of the presence of hepatotoxic pyrrolizidine alkaloids (PAs). A systematic toxicological study is thus required to verify this public concern. In the present article, we report, for the first time, that S. SCANDENS contains nine hepatotoxic PAs with a content of 6.95-7.19 microg/g. At a dose equivalent to the daily intake recommended by the Pharmacopoeia of China, the total content of toxic PAs in Qianliguang was determined to be 3.48 microg/kg/day, which is far below the lowest dose to cause hepatotoxicity (15 microg/kg/day) suggested by the International Program on Chemical Safety. No significant hepatotoxic effects were observed in rats fed with the extract at this human-equivalent dose for 14 consecutive days. However, a single overdose of the herbal water extract (6 g/kg), which was about 8-fold higher than the recommended dose, produced typical PA-induced hepatotoxicity in rats. Therefore, appropriate dosage guidelines should be implemented for the herbal industry, for export/import retailers, and for herbal medicine practitioners to ensure the safe and beneficial use of these herbal medicines. PMID- 19296429 TI - Effectiveness of an innovative prototype subtracted diversity array (SDA) for fingerprinting plant species of medicinal importance. AB - The accurate identification of medicinal plants is becoming increasingly important due to reported concerns about purity, quality and safety. The previously developed prototype subtracted diversity array (SDA) had been validated for the ability to distinguish clade-level targets in a phylogenetically accurate manner. This study represents the rigorous investigation of the SDA for genotyping capabilities, including the genotyping of plant species not included during the construction of the SDA, as well as to lower classification levels including family and species. The results show that the SDA, in its current form, has the ability to accurately genotype species not included during SDA development to clade level. Additionally, for those species that were included during SDA development, genotyping is successful to the family level, and to the species level with minor exceptions. Twenty polymorphic SDA features were sequenced in a first attempt to characterize the polymorphic DNA between species, which showed that transposon-like sequences may be valuable as polymorphic features to differentiate angiosperm families and species. Future refinements of the SDA to allow more sensitive genotyping are discussed with the overall goal of accurate medicinal plant identification in mind. PMID- 19296430 TI - New cytotoxic lupane triterpenes from Perrottetia arisanensis. AB - Eight new lupane triterpenes, including 7beta-cis-coumaroylbetulinic acid (1), 7beta-trans-coumaroylbetulinic acid (2), 7beta-cis-coumaroyl-3-epi-betulinic acid (3), 7beta-trans-coumaroyl-3-epi-betulinic acid (4), 7beta-cis-coumaroylbetulonic acid (5), 7beta-trans-coumaroylbetulonic acid (6), 7beta-hydroxybetulinaldehyde (7) and 28-norlup-20(29)-ene-3alpha,17beta-diol (8), together with fifteen known compounds were isolated from the bioactive methanol extract of the stems of Perrottetia arisanensis. The structures of the new compounds were elucidated by spectroscopic and HR-ESI-MS analysis. All new compounds were evaluated for their cytotoxicity against six human cancer cell lines. Among them, lupane triterpene coumaroyl esters 1-6 showed moderate cytotoxicity with IC (50) values ranging from 3.75 to 21.29 microM. This is the first report for lupane triterpenes with a phenylpropane moiety substituted at C-7. PMID- 19296431 TI - Cytotoxic phenanthroindolizidine alkaloids from the roots of Ficus septica. AB - Activity-monitored fractionation of the CHCl3 solubles of the MeOH extract of the roots of Ficus septica using cytotoxicity assay led to the isolation of twenty phenanthroindolizidine alkaloids, including ten new ficuseptines E-N and ten known compounds. The cytotoxic effects of some of the isolates were tested on gastric adenocarcinoma (NUGC) and nasopharyngeal carcinoma (HONE-1) cell lines and the results demonstrated that, with the exception of dehydrotylophorine, all the tested compounds displayed pronounced cytotoxic activity with significant cell growth inhibitory effects. In addition, 13a R-antofine was found to be highly effective with ED(50) values of < 0.1 microg/mL against L-1210, P-388, A 549 and HCT-8 cell lines in another assay. PMID- 19296432 TI - Phenolic compounds in the leaves of Populus ussuriensis and their antioxidant activities. AB - Two new phenolic glucosides [isograndidentatin A (1) isograndidentatin B (2)], 5 known phenolic glucosides [grandidentatin (3), salireposide (4), populoside (5), populoside A (6), and salicortin (7)], and 2 known phenolic acids [P-coumaric acid (8) and caffeic acid (9)] were isolated from the leaves of Populus ussuriensis. Structure elucidation of 1 and 2 was achieved through extensive spectroscopic techniques. Compounds 1-6 and 9 showed significant antioxidant activities, which were evaluated by the DPPH radical-scavenging method (IC(50) values of 6.68, 6.61, 6.75, 6.84, 6.76, 6.79, and 5.92 microM, respectively) and the ABTS .+ radical-scavenging system (TEAC values of 1.21, 1.28, 1.26, 1.05, 1.69, 1.60, and 2.00 mM, respectively). PMID- 19296433 TI - Composition of Indigo naturalis. AB - A proposal for a European Pharmacopoeia monograph concerning Indigo naturalis has recently been published, whereby the indigo (1) and indirubin (2) content should be determined by HPLC-UV. This method was tested, but problems were seen with the dosage of indigo due to poor solubility. A quantitative assay for indigo based on (1)H-NMR was developed as an alternative. The HPLC and qNMR assays were compared with eight Indigo naturalis samples. The HPLC assay consistently gave much lower indigo concentrations because solubility was the limiting factor in sample preparation. In one sample, sucrose was identified by (1)H-NMR as an organic additive. Simple wet chemistry assays for undeclared additives such as sugars and starch were tested with artificially spiked Indigo naturalis samples to establish their limits of detection, and sulfate ash determinations were carried out in view of a better assessment of Indigo naturalis in a future European monograph. PMID- 19296434 TI - Today's biotech industry in India. PMID- 19296435 TI - Profiles of four top biotech companies in India. PMID- 19296436 TI - Basic and applied research in India: present and future. PMID- 19296437 TI - Biotech regulation in India: problems and promises. PMID- 19296438 TI - Spotlight on botanics in India: characterization of Thar desert plants. PMID- 19296440 TI - An Indian effort towards affordable drugs: "generic to designer drugs". PMID- 19296441 TI - O-acetylated sialic acids: multifaceted role in childhood acute lymphoblastic leukaemia. AB - Childhood acute lymphoblastic leukaemia (ALL), a malignant transformation of the lymphoblasts, is highly responsive to chemotherapy. However, due to certain inadequacy in detection of minimal residual disease (MRD), relapse is a common phenomenon. To address this question, the present review deals with the induction of an unique O-acetyl derivative of sialic acid on a few disease-associated glycoproteins and glycolipids at the onset of childhood ALL, a finding of our group in the last decade. This information has been successfully utilized for diagnosis and prognosis of the disease. Existing literature is included for comparison. Additionally, cell surface overexpression of 9-O-acetylated sialoglycoproteins and antibodies against them present in patients' sera aid the survival of the malignant lymphoblasts and suggest a multifaceted role played by these molecules. Taken together, monitoring these molecules helps not only in unravelling the biology of this paediatric malignancy but also in personalizing the treatment strategies for the betterment of the patient population. PMID- 19296442 TI - Reversible acetylation of chromatin: implication in regulation of gene expression, disease and therapeutics. AB - The eukaryotic genome is a highly dynamic nucleoprotein complex that is comprised of DNA, histones, nonhistone proteins and RNA, and is termed as chromatin. The dynamic of the chromatin is responsible for the regulation of all the DNA templated phenomena in the cell. Several factors, including the nonhistone chromatin components, ATP-dependent remodeling factors and the chromatin modifying enzymes, mediate the combinatorial post-translational modifications that control the chromatin fluidity and, thereby, the cellular functions. Among these modifications, reversible acetylation plays a central role in the highly orchestrated network. The enzymes responsible for the reversible acetylation, the histone acetyltransferases (HATs) and histone deacetylases (HDACs), not only act on histone substrates but also on nonhistone proteins. Dysfunction of the HATs/HDACs is associated with various diseases like cancer, diabetes, asthma, cardiac hypertrophy, retroviral pathogenesis and neurodegenerative disorders. Therefore, modulation of these enzymes is being considered as an important therapeutic strategy. Although substantial progress has been made in the area of HDAC inhibitors, we have focused this review on the HATs and their small-molecule modulators in the context of disease and therapeutics. Recent discoveries from different groups have established the involvement of HAT function in various diseases. Furthermore, several new classes of HAT modulators have been identified and their biological activities have also been reported. The scaffold of these small molecules can be used for the design and synthesis of better and efficient modulators with superior therapeutic efficacy. PMID- 19296443 TI - Efficacy of recombinant anthrax vaccine against Bacillus anthracis aerosol spore challenge: preclinical evaluation in rabbits and Rhesus monkeys. AB - This report describes the immunogenicity and protective efficacy of Escherichia coli-expressed recombinant protective antigen (rPA) in New Zealand White rabbits and Rhesus Macaques against an aerosol challenge with Bacillus anthracis spores (IVRI strain, tox+cap+). A dose-ranging study was performed in which it became evident that the level of anti-PA IgG and toxin-neutralizing antibody titer was directly proportional to the dose of rPA administered. However, the onset time of primary and secondary immune response was not dependent on the dosage. Revaccination of primed animals with the same threshold dose yielded a robust and rapid secondary response. Quantitative differences in peak titers were obtained for both the animal models, in addition to qualitative differences in the immune kinetics. In spite of a weak priming response, the secondary response in rabbits peaked earlier than that in macaques once the booster dose was administered. However, evaluation of the post-challenge quantitative anti-rPA ELISA titer measurements indicated higher titers for non-human primates as compared to the lagomorphs. Importantly, 100% protection was seen for the dosage groups that received > or = 25 microg rPA, following a challenge against a target dose of 1000 LD(50) of aerosolized spores of Bacillus anthracis. PMID- 19296445 TI - Editorial: Biotech in India--history, present and promises. PMID- 19296449 TI - Functional screening of adrenergic receptors by measuring intracellular calcium using the FlexStation scanning fluorimeter. AB - In this study we test whether functional screening of compounds to adrenergic G protein-coupled receptors (GPCRs) would provide data that correlated significantly with radiolabeled binding data, thereby permitting researchers to replace expensive radioligand-binding experiments with non-radioactive screening. An increase in intracellular calcium levels represents an important second messenger signal for several recombinant GPCRs. In this study, we describe the affinities of three alpha adrenoceptor antagonists (terazosin, tamsulosin and alfuzosin), determined by monitoring the changes in intracellular calcium levels and comparing them with their radioligand-binding affinities. In addition to determining the functional affinities of the three alpha adrenoceptor antagonists, we evaluate their binding at two alpha adrenoceptor subtypes and optimized the assay for high-throughput screening. PMID- 19296450 TI - Bioremediation of phenol by a novel partitioning bioreactor using cow dung microbial consortia. AB - A bioreactor has been designed and developed for partitioning of aqueous and organic phases with a provision for aeration and stirring, a cooling system and a sampling port. The potential of a cow dung microbial consortium has been assessed for bioremediation of phenol in a single-phase bioreactor and a two-phase partitioning bioreactor. The advantages of the two-phase partitioning bioreactor are discussed. The Pseudomonas putida IFO 14671 has been isolated, cultured and identified from the cow dung microbial consortium as a high-potential phenol degrader. The methods developed in this study present an advance in bioremediation techniques for the biodegradation of organic compounds such as phenol using a bioreactor. We have also demonstrated the potential of microorganisms from cow dung as a source of biomass. PMID- 19296452 TI - Biotechnology in India: the beginnings. PMID- 19296453 TI - Development and exploitation of DNA fingerprinting technology in India. PMID- 19296454 TI - Molecular approaches for the fixation of plant hybrid vigor. PMID- 19296455 TI - The molecular evolution of HIV-1 protease simulated at atomic detail. AB - Progress in understanding protein folding allows to simulate, with atomic detail, the evolution of amino-acid sequences folding to a given native conformation. A particularly attractive example is the HIV-1 protease, main target of therapies to fight AIDS, which under drug pressure is able to develop resistance within few months from the starting of therapy. By comparing the results of simulations of the evolution of the protease with the corresponding proteomic data, one can approximately determine the value of the associated evolution pressure under which the enzyme has become and, as a consequence, map out the energy landscape in sequence space of the HIV-1 protease. It is found that there are several families of sequences folding to the native conformations of the enzyme. Each of these families are characterized by different sets of highly conserved ("hot") amino acids which play a critical role in the folding and stability of the protease. There are two main possibilities for the virus to move from one family to a different one: (a) in a single generation, through the concerted mutations of the hot amino acids, a highly unlikely event, (b) through a folding path (if it exists), again a very improbable event. In fact, the number of generations needed by the virus to change stepwise its sequence from one family to another is astronomically large. These results point to the "hot" segments of the protease as promising targets for a nonconventional inhibition strategy, likely not to create resistance. PMID- 19296456 TI - Membrane insertion of the FYVE domain is modulated by pH. AB - The FYVE domain associates with phosphatidylinositol 3-phosphate [PtdIns(3)P] in membranes of early endosomes and penetrates bilayers. Here, we detail principles of membrane anchoring and show that the FYVE domain insertion into PtdIns(3)P enriched membranes and membrane-mimetics is substantially increased in acidic conditions. The EEA1 FYVE domain binds to POPC/POPE/PtdIns(3)P vesicles with a Kd of 49 nM at pH 6.0, however associates approximately 24 fold weaker at pH 8.0. The decrease in the affinity is primarily due to much faster dissociation of the protein from the bilayers in basic media. Lowering the pH enhances the interaction of the Hrs, RUFY1, Vps27p and WDFY1 FYVE domains with PtdIns(3)P containing membranes in vitro and in vivo, indicating that pH-dependency is a general function of the FYVE finger family. The PtdIns(3)P binding and membrane insertion of the FYVE domain is modulated by the two adjacent His residues of the R(R/K)HHCRXCG signature motif. Mutation of either His residue abolishes the pH sensitivity. Both protonation of the His residues and nonspecific electrostatic contacts stabilize the FYVE domain in the lipid-bound form, promoting its penetration and increasing the membrane residence time. PMID- 19296457 TI - Role of ubiquitin-proteasome proteolysis in muscle fiber destruction in experimental chloroquine-induced myopathy. AB - Previous studies have documented the presence of rimmed vacuoles, atrophic fibers, and increased lysosomal cathepsin activity in skeletal muscle from animal models of chloroquine-induced myopathy, suggesting that muscle fibers in this type of myopathy may be degraded via the lysosomal-proteolysis pathway. Given recent evidence of abnormal ubiquitin accumulation in rimmed vacuoles, in this study we chose to examine the significance of the ubiquitin-proteasome proteolytic system in the process of muscle fiber destruction in experimental chloroquine myopathy. Expression of ubiquitin, 26S proteasome proteins, and ubiquitin ligases, such as muscle-specific RING finger-1 (MuRF-1) and atrogin 1/muscle atrophy F-box protein (MAFbx), was analyzed in innervated and denervated rat soleus muscles after treatment with either saline or chloroquine. Abnormal accumulation of rimmed vacuoles was observed only in chloroquine-treated denervated muscles. Ubiquitin and proteasome immunostaining, and ubiquitin, MuRF 1, and atrogin-1/MAFbx mRNAs were significantly increased in denervated soleus muscles from saline- and chloroquine-treated rats when compared with contralateral innervated muscles. Further, ubiquitin and ubiquitin ligase mRNA levels were higher in denervated muscles from chloroquine-treated rats when compared with saline-treated rats. These data demonstrate increased proteasomes and ubiquitin in denervated muscles from chloroquine-treated rats and suggest that the ubiquitin-proteasome proteolysis pathway as well as the lysosomal proteolysis pathway mediate muscle fiber destruction in experimental chloroquine myopathy. PMID- 19296458 TI - The deep breathing test: median-based expiration-inspiration difference is the measure of choice. AB - Heart rate variability (HRV) has become an important parameter for the assessment of autonomic function in many areas of medicine. In particular, respiratory sinus arrhythmia measured during the deep breathing test (DBT) is often used. Results are usually expressed in common time-domain parameters. A "most preferred measure" has not yet been identified. We investigated the sensitivity of the DBT to the following anomalies: in-test variance; shifts of mean heart rate; premature ventricular contractions; and breathing rate deviations. Frequency and magnitude of the anomalies were determined in a set of real DBTs (n=514) and transferred to computer simulations to mimic realistic conditions. The sensitivity of standard deviation, mean circular resultant (MCR), root mean square of successive differences (RMSSD), and four types of expiration inspiration (E-I) difference were quantified statistically. Median-based E-I differences, E-I ratio, and MCR were most resistant to the anomalies. E-I difference derived by median values should be used preferentially, providing the highest precision and independence from heart rate. PMID- 19296459 TI - CMAP decrement in ALS. PMID- 19296460 TI - Dynametric assessment in CIDP. PMID- 19296461 TI - Can the Distress Thermometer be improved by additional mood domains? Part II. What is the optimal combination of Emotion Thermometers? AB - PURPOSE: To examine the added value of an algorithmic combination of visual analogue thermometers compared with the Distress Thermometer (DT) when attempting to detect depression, anxiety or distress in early cancer. METHODS: We report Classification and Regression Tree and logistic regression analyses of the new five-domain Emotion Thermometers tool. This is a combination of five visual analogue scales in the form of four mood domains (distress, anxiety, depression, anger) as well as need for help. 130 patients attending for their first chemotherapy treatment were assessed. We calculated optimal accuracy for each domain alone and in combination against several criterion standards. RESULTS: When attempting to diagnose depression the Depression Thermometer (DepT) used alone was the optimal approach, but when attempting to detect broadly defined distress or anxiety then a combination of thermometers was most accurate. The DepT was significantly more accurate in detecting depression than the DT. For broadly defined distress a combination of depression, anger and help thermometers was more accurate than the DT alone. For anxiety, while the anxiety thermometer (AnxT) improves upon the DT alone, a combination of the DepT and AnxT are optimal. In each case the optimal strategy allowed the detection of at least an additional 9% of individuals. However, combinations are more laborious to score. In settings where the simplest possible option is preferred the most accurate single thermometer might be preferable as a first stage assessment. CONCLUSION: The DT can be improved by specific combinations of simple thermometers that incorporate depression, anxiety, anger and help. PMID- 19296462 TI - Can the Distress Thermometer be improved by additional mood domains? Part I. Initial validation of the Emotion Thermometers tool. AB - PURPOSE: To examine the value of a new screening instrument in a visual-analogue format. METHODS: We report the design and validation of a new five-dimensional tool called the Emotion Thermometers (ET). This is a combination of five visual analogue scales in the form of four predictor domains (distress, anxiety, depression, anger) and one outcome domain (need for help). Between March and August 2007, 130 patients attending the chemotherapy suite for their first chemotherapy treatment were asked to complete several questionnaires with validation for distress, anxiety and depression. RESULTS: Of 81 with low distress on the Distress Thermometer (DT), 51% recorded emotional difficulties on the new ET tool, suggesting added value beyond the DT alone. Of those with a broadly defined emotional complication, 93.3% could be identified using the Anxiety Thermometer (AnxT) alone, compared with 54.4% who would be recognized using the DT alone. Using a cut-off of 3v4 on all thermometers against the total Hospital Anxiety and Depression Scale (HADS) score (cut-off 14v15), the optimal thermometer was the Anger Thermometer (sensitivity 61%, specificity 92%). Against HADS anxiety scale, the optimal thermometer was AnxT (sensitivity 92%, specificity 61%) and against the HADS depression scale, the optimal thermometer was the Depression Thermometer (DepT; sensitivity 60%, specificity 78%). Finally, against DSM-IV major depression, the optimal thermometer was the DepT (sensitivity 80%, specificity 79%). Further improvements may be possible by using a combination of thermometers or by repeating the screen. CONCLUSION: The diagnostic accuracy of the DT can be improved by the inclusion of simple addition linear domains without substantially increasing the time needed to apply the test. PMID- 19296463 TI - Effects of antiepileptic drugs on lipids, homocysteine, and C-reactive protein. AB - OBJECTIVE: The widely prescribed anticonvulsants phenytoin and carbamazepine are potent inducers of cytochrome P450 enzymes, which are involved in cholesterol synthesis. We sought to determine whether these drugs have an effect on cholesterol and other serological markers of vascular risk. METHODS: We recruited 34 epilepsy patients taking carbamazepine or phenytoin in monotherapy whose physicians had elected to change treatment to one of the noninducing anticonvulsants lamotrigine or levetiracetam. Fasting blood samples were obtained both before and 6 weeks after the switch to measure serum lipid fractions, lipoprotein(a), C-reactive protein, and homocysteine. A comparator group of 16 healthy subjects underwent the same serial studies. RESULTS: In the epilepsy patients, switch from either phenytoin or carbamazepine produced significant declines in total cholesterol (-24.8 mg/dl), atherogenic (non-high-density lipoprotein) cholesterol (-19.9 mg/dl), triglycerides (-47.1mg/dl) (all p < 0.0001), and C-reactive protein (-31.4%; p = 0.027). Patients who stopped taking carbamazepine also had a 31.2% decline in lipoprotein(a) level (p = 0.0004), whereas those taken off phenytoin had a decrease in homocysteine level (-1.7 micromol/L; p = 0.005). All of these changes were significant when compared with those seen in healthy subjects (p < 0.05). Results were similar whether patients were switched to lamotrigine or levetiracetam. INTERPRETATION: Switching epilepsy patients from the enzyme-inducers carbamazepine or phenytoin to the noninducing drugs levetiracetam or lamotrigine produces rapid and clinically significant amelioration in several serological markers of vascular risk. These findings suggest that phenytoin and carbamazepine may substantially increase the risk for cardiovascular and cerebrovascular disease. PMID- 19296464 TI - Divergent and point-to-point connections in the commissural pathway between the inferior colliculi. AB - The commissure of the inferior colliculus interconnects the left and right sides of the auditory midbrain and provides the final opportunity for interaction between the two sides of the auditory pathway at the subcortical level. Although the functional properties of the commissure are beginning to be revealed, the topographical organization of its connections is unknown. A combination of neuroanatomical tracing studies, 3D reconstruction, and neuronal density maps was used to study the commissural connections in rat. The results demonstrate that commissural neurons in the central nucleus of the inferior colliculus send a divergent projection to the equivalent frequency-band laminae in the central nucleus and dorsal and lateral cortices on the opposite side. The density of this projection, however, is weighted toward a point that matches the position of the tracer injection; consistent with a point-to-point emphasis in the wiring pattern. In the dorsal cortex of the inferior colliculus there may be two populations of neurons that project across the commissure, one projecting exclusively to the frequency-band laminae in the central nucleus and the other projecting diffusely to the dorsal cortex. Neurons in the lateral cortex of the inferior colliculus make only a very weak contribution to the commissural pathway. The point-to-point pattern of connections permits interactions between specific regions of corresponding frequency-band laminae, whereas the divergent projection pattern could subserve integration across the lamina. PMID- 19296465 TI - Loss of protein kinase Cbeta function protects mice against diet-induced obesity and development of hepatic steatosis and insulin resistance. AB - Obesity is an energy balance disorder in which intake is greater than expenditure, with most excess calories stored as triglyceride (TG). We previously reported that mice lacking the beta-isoform of protein kinase C (PKCbeta), a diacylglycerol- and phospholipid-dependent kinase, exhibit marked reduction in the whole body TG content, including white adipose tissue (WAT) mass. To investigate the role of this signaling kinase in metabolic adaptations to severe dietary stress, we studied the impact of a high-fat diet (HFD) on PKCbeta expression and the effect of PKCbeta deficiency on profound weight gain. We report herein that HFD selectively increased PKCbeta expression in obesity-prone C57BL/6J mice, specifically in WAT; the expression levels were little or unchanged in the liver, muscle, kidney, and heart. Basal PKCbeta expression was also found to be elevated in WAT of obese ob/ob mice. Remarkably, mice lacking PKCbeta were resistant to HFD-induced obesity, showing significantly reduced WAT and slightly higher core body temperatures. Unlike lean lipodystrophic mouse models, these mice did not have fatty livers, nor did they exhibit insulin resistance. Moreover, PKCbeta(-/-) mice exhibited changes in lipid metabolism gene expression, and such alterations were accompanied by significant changes in serum adipokines. These observations suggest that PKCbeta deficiency induced a unique metabolic state congruous with obesity resistance, thus raising the possibility that dysregulation of PKCbeta expression could contribute to dietary fat-induced obesity and related disorders. PMID- 19296466 TI - Taurine supplementation prevents ethanol-induced decrease in serum adiponectin and reduces hepatic steatosis in rats. AB - Chronic ethanol feeding decreases expression of adiponectin by adipocytes and circulating adiponectin. Adiponectin treatment during chronic ethanol feeding prevents liver injury in mice. Chronic ethanol feeding also increases oxidative and endoplasmic reticulum (ER) stress in adipose tissue. Here we tested the hypothesis that supplemental taurine, an amino acid that functions as a chemical chaperone/osmolyte and enhances cellular antioxidant activity, would prevent ethanol-induced decreases in adiponectin expression and attenuate liver injury. Serum adiponectin concentrations decreased as early as 4 to 7 days after feeding rats a 36% ethanol diet. This rapid decrease was associated with increased oxidative, but not ER, stress in subcutaneous adipose tissue. Taurine prevented ethanol-induced oxidative stress and increased inflammatory cytokine expression in adipose tissue. Ethanol feeding also rapidly decreased expression of transcription factors regulating adiponectin expression (CCAAT/enhancer binding protein alpha; peroxisome proliferator-activated receptor alpha/gamma) in subcutaneous adipose tissue. Taurine prevented the ethanol-induced decrease in CCAAT/enhancer binding protein alpha and peroxisome proliferator-activated receptor alpha, normalizing adiponectin messenger (m)RNA and serum adiponectin concentrations. In the liver, taurine prevented ethanol-induced oxidative stress and attenuated tumor necrosis factor alpha expression and steatosis, at least in part, by increasing expression of genes involved in fatty acid oxidation. CONCLUSION: In subcutaneous adipose tissue, taurine decreased ethanol-induced oxidative stress and cytokine expression, as well as normalized expression of adiponectin mRNA. Taurine prevented ethanol-induced decreases in serum adiponectin; normalized adiponectin was associated with a reduction in hepatic oxidative stress, tumor necrosis factor alpha expression, and steatosis. Taken together, these data demonstrate that taurine has important protective effects against ethanol-induced tissue injury in both adipose and liver tissue. PMID- 19296467 TI - Endotoxin uptake in mouse liver is blocked by endotoxin pretreatment through a suppressor of cytokine signaling-1-dependent mechanism. AB - The liver is the main organ that clears lipopolysaccharide (LPS) and hepatocytes are a major cell-type involved in LPS uptake. LPS tolerance, or desensitization, is important in negative regulation of responses to LPS, but little is known about its mechanisms in hepatocytes. Primary isolated C57BL/6 hepatocytes, and liver in vivo, internalized fluorescent LPS, and this was dependent on Toll-like receptor 4 (TLR4) at the cell surface but not on TLR4-TIR signaling through MyD88. LPS clearance from plasma was also TLR4-dependent. Pretreatment of C57BL/6 hepatocytes with LPS prevented uptake of LPS 24 hours later and this LPS-mediated suppression was dependent on TLR4 signaling through MyD88. Many regulators of TLR4 signaling have been identified and implicated in LPS desensitization, including suppressor of cytokine signaling 1 (SOCS1). SOCS1 mRNA and protein expression increased after LPS stimulation in hepatocytes and in whole liver. LPS uptake in hepatocytes and liver was significantly reduced following infection with adenoviral vectors overexpressing SOCS1. Similarly, inhibition of SOCS1 using small interfering (si)RNA-mediated knockdown prevented LPS desensitization in hepatocytes. SOCS1 is known to interact with Toll/IL-1 receptor associated protein (TIRAP) and cause TIRAP ubiquitination and degradation, which regulates TLR signaling. We have also shown previously that TIRAP regulates LPS uptake in hepatocytes. SOCS1 coimmunoprecipitated with TIRAP in wild type hepatocyte cell lysates up to 8 hours after LPS stimulation, but not at later times. In the same samples, ubiquitinated TIRAP was detected after 4 hours and up to 8 hours after LPS stimulation, but not at later times. CONCLUSION: These data indicate hepatocytes are desensitized by LPS in a TLR4 signaling-dependent manner. LPS induced SOCS1 upregulation increases degradation of TIRAP and prevents subsequent LPS uptake. The exploitation of these mechanisms of LPS desensitization in the liver may be important in future sepsis therapies. PMID- 19296468 TI - MicroRNA-21 is overexpressed in human cholangiocarcinoma and regulates programmed cell death 4 and tissue inhibitor of metalloproteinase 3. AB - Cholangiocarcinomas (CCAs) are aggressive cancers, with high mortality and poor survival rates. Only radical surgery offers patients some hope of cure; however, most patients are not surgical candidates because of late diagnosis secondary to relatively poor accuracy of diagnostic means. MicroRNAs (miRs) are involved in every cancer examined, but they have not been evaluated in primary CCA. In this study, miR arrays were performed on five primary CCAs and five normal bile duct specimens (NBDs). Several miRs were dysregulated and miR-21 was overexpressed in CCAs. miR-21 differential expression in these 10 specimens was verified by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). To validate these findings, qRT-PCR for miR-21 was then performed on 18 additional primary CCAs and 12 normal liver specimens. MiR-21 was 95% sensitive and 100% specific in distinguishing between CCA and normal tissues, with an area under the receiver operating characteristic curve of 0.995. Inhibitors of miR-21 increased protein levels of programmed cell death 4 (PDCD4) and tissue inhibitor of metalloproteinases 3 (TIMP3). Notably, messenger RNA levels of TIMP3 were significantly lower in CCAs than in normals. CONCLUSIONS: MiR-21 is overexpressed in human CCAs. Furthermore, miR-21 may be oncogenic, at least in part, by inhibiting PDCD4 and TIMP3. Finally, these data suggest that TIMP3 is a candidate tumor suppressor gene in the biliary tree. PMID- 19296469 TI - Relation between liver progenitor cell expansion and extracellular matrix deposition in a CDE-induced murine model of chronic liver injury. AB - In chronic liver injury, liver progenitor cells (LPCs) proliferate in the periportal area, migrate inside the lobule, and undergo further differentiation. This process is associated with extracellular matrix (ECM) remodeling. We analyzed LPC expansion and matrix accumulation in a choline-deficient, ethionine supplemented (CDE) model of LPC proliferation. After day 3, CDE induced collagen deposits in the periportal area. Expansion of LPCs as assessed by increased number of cytokeratin 19 (CK19)-positive cells was first observed at day 7, while ECM accumulated 10 times more than in controls. Thereafter, LPCs and ECM increased in parallel. Furthermore, ECM not only accumulates prior to the increase in number of LPCs, but is also found in front of LPCs along the porto venous gradient of lobular invasion. Double immunostaining revealed that LPCs are embedded in ECM at all times. Moreover, LPCs infiltrating the liver parenchyma are chaperoned by alpha-smooth muscle actin (alpha-SMA)-positive cells. Gene expression analyses confirmed these observations. The expression of CK19, alpha fetoprotein, E-cadherin, and CD49f messenger RNA (mRNA), largely overexpressed by LPCs, significantly increased between day 7 and day 10. By contrast, at day 3 there was a rapid burst in the expression of components of the ECM, collagen I and laminin, as well as in alpha-SMA and connective tissue growth factor expression. CONCLUSION: Our data demonstrate that, in a CDE model, ECM deposition and activation of matrix-producing cells occurred as an initial phase, prior to LPC expansion, and in front of LPCs along the porto-venous gradient of lobular invasion. Those observations may reveal a fundamental role for the established hepatic microenvironment or niche during the process of activation and differentiation of liver progenitor cells. PMID- 19296470 TI - MicroRNA-122, a tumor suppressor microRNA that regulates intrahepatic metastasis of hepatocellular carcinoma. AB - MicroRNAs (miRNAs), which are inhibitors of gene expression, participate in diverse biological functions and in carcinogenesis. In this study, we show that liver-specific microRNA-122 (miR-122) is significantly down-regulated in liver cancers with intrahepatic metastasis and negatively regulates tumorigenesis. Restoration of miR-122 in metastatic Mahlavu and SK-HEP-1 cells significantly reduced in vitro migration, invasion, and anchorage-independent growth as well as in vivo tumorigenesis, angiogenesis, and intrahepatic metastasis in an orthotopic liver cancer model. Because an inverse expression pattern is often present between an miRNA and its target genes, we used a computational approach and identified multiple miR-122 candidate target genes from two independent expression microarray datasets. Thirty-two target genes were empirically verified, and this group of genes was enriched with genes regulating cell movement, cell morphology, cell-cell signaling, and transcription. We further showed that one of the miR-122 targets, ADAM17 (a disintegrin and metalloprotease 17) is involved in metastasis. Silencing of ADAM17 resulted in a dramatic reduction of in vitro migration, invasion, in vivo tumorigenesis, angiogenesis, and local invasion in the livers of nude mice, which is similar to that which occurs with the restoration of miR-122. CONCLUSION: Our study suggests that miR 122, a tumor suppressor microRNA affecting hepatocellular carcinoma intrahepatic metastasis by angiogenesis suppression, exerts some of its action via regulation of ADAM17. Restoration of miR-122 has a far-reaching effect on the cell. Using the concomitant down-regulation of its targets, including ADAM17, a rational therapeutic strategy based on miR-122 may prove to be beneficial for patients with hepatocellular carcinoma. PMID- 19296471 TI - Bile acids: trying to understand their chemistry and biology with the hope of helping patients. AB - An informal review of the author's five decades of research on the chemistry and biology of bile acids in health and disease is presented. The review begins with a discussion of bile acid structure and its remarkable diversity in vertebrates. Methods for tagging bile acids with tritium for metabolic or transport studies are summarized. Bile acids solubilize polar lipids in mixed micelles; progress in elucidating the structure of the mixed micelle is discussed. Extensive studies on bile acid metabolism in humans have permitted the development of physiological pharmacokinetic models that can be used to simulate bile acid metabolism. Consequences of defective bile acid biosynthesis and transport have been clarified, and therapy has been developed. Methods for measuring bile acids have been improved. The rise and fall of medical and contact dissolution of cholesterol gallstones is chronicled. Finally, principles of therapy with bile acid agonists and antagonists are given. Advances in understanding bile acid biology and chemistry have helped to improve the lives of patients with hepatobiliary or digestive disease. PMID- 19296472 TI - Myeloma cells exhibit an increase in proteasome activity and an enhanced response to proteasome inhibition in the bone marrow microenvironment in vivo. AB - The proteasome inhibitor bortezomib has a striking clinical benefit in patients with multiple myeloma. It is unknown whether the bone marrow microenvironment directly contributes to the dramatic response of myeloma cells to proteasome inhibition in vivo. We have used the well-characterized 5TGM1 murine model of myeloma to investigate myeloma growth within bone and response to the proteasome inhibitor bortezomib in vivo. Myeloma cells freshly isolated from the bone marrow of myeloma-bearing mice were found to have an increase in proteasome activity and an enhanced response to in vitro proteasome inhibition, as compared with pre inoculation myeloma cells. Treatment of myeloma-bearing mice with bortezomib resulted in a greater reduction in tumor burden when the myeloma cells were located within the bone marrow when compared with extra-osseous sites. Our results demonstrate that myeloma cells exhibit an increase in proteasome activity and an enhanced response to bortezomib treatment when located within the bone marrow microenvironment in vivo. PMID- 19296473 TI - Gaucher disease: resetting the clinical and scientific agenda. PMID- 19296474 TI - Distinct roles of neuropilin 1 signaling for radial and tangential extension of callosal axons. AB - Cortical excitatory neurons migrate from their origin in the ventricular zone (VZ) toward the pial surface. During migration, these neurons exhibit a stellate shape in the intermediate zone (IZ), transform into bipolar cells, and then initiate radial migration, extending a trailing process, which may lead to an axon. Here we examined the role of neuropilin 1 (NRP1) in these developmental events. Both NRP1 mRNA and protein were highly expressed in the IZ, where stellate-shaped cells were located. DiI labeling experiments showed that neuronal migration occurred normally in Nrp1 mutant mice up to embryonic day (E) 14.5, the latest day to which the mutant survives, with only subtle axonal defasciculation. However, interference with Nrp1 signaling at a later stage caused pathfinding errors: when a dominant negative form of Nrp1 was electroporated into the cortical VZ cells at E12.5 or E15.5 and examined perinatally, guidance errors were found in tangential axonal extension toward the midline. In contrast, no significant effect was noted on the migration of cortical excitatory neurons. These findings indicate that NRP1 plays an important role in the guidance of callosal axons originating from cortical excitatory neurons but does not support a role in their migration. Moreover, insofar as radial axonal extension within the cortical plate was unaffected, the present findings imply that molecular mechanisms for the axonal extension of excitatory neurons within the cortical plate are distinct from those in the white matter. PMID- 19296475 TI - Binge-like postnatal alcohol exposure triggers cortical gliogenesis in adolescent rats. AB - The long-term effects of binge-like postnatal alcohol exposure on cell proliferation and differentiation in the adolescent rat neocortex were examined. Unlike the hippocampal dentate gyrus, where proliferation of progenitors results primarily in addition of granule cells in adulthood, the vast majority of newly generated cells in the intact mature rodent neocortex appear to be glial cells. The current study examined cytogenesis in the motor cortex of adolescent and adult rats that were exposed to 5.25 g/kg/day of alcohol on postnatal days (PD) 4 9 in a binge manner. Cytogenesis was examined at PD50 (through bromodeoxyuridine [BrdU] labeling) and survival of these newly generated cells was evaluated at PD80. At PD50, significantly more BrdU-positive cells were present in the motor cortex of alcohol-exposed rats than controls. Confocal analysis revealed that the majority (>60%) of these labeled cells also expressed NG2 chondroitin sulfate proteoglycan (NG2 glia). Additionally, survival of these newly generated cortical cells was affected by neonatal alcohol exposure, based on the greater reduction in the number of BrdU-labeled cells from PD50 to PD80 in the alcohol-exposed animals compared to controls. These findings demonstrate that neonatal alcohol exposure triggers an increase in gliogenesis in the adult motor cortex. PMID- 19296477 TI - Proteomic analysis of striatal neuronal cell cultures after morphine administration. AB - Using primary neuronal cell culture assays, combined with 2-D gel electrophoresis and capillary LC-MS, we identified differences in proteomes between control and morphine-treated cells. Statistically significant differences were observed among 26 proteins. Nineteen of them were up-regulated, while seven were down-regulated in morphine-treated cell populations. The identified proteins belong to classes involved in energy metabolism, associated with oxidative stress, linked with protein biosynthesis, cytoskeletal ones, and chaperones. The detected proteins demand further detailed studies of their biological roles in morphine addiction. It is crucial to confirm observed processes in vivo in order to reveal the nature and importance of the biological effect of proteome changes after morphine administration. Further investigations may lead to the discovery of new proteome based effects of morphine on living organisms. PMID- 19296476 TI - Altered cutaneous nerve regeneration in a simian immunodeficiency virus / macaque intracutaneous axotomy model. AB - To characterize the regenerative pattern of cutaneous nerves in simian immunodeficiency virus (SIV)-infected and uninfected macaques, excisional axotomies were performed in nonglabrous skin at 14-day intervals. Samples were examined after immunostaining for the pan-axonal marker PGP 9.5 and the Schwann cell marker p75 nerve growth factor receptor. Collateral sprouting of axons from adjacent uninjured superficial dermal nerve bundles was the initial response to axotomy. Both horizontal collateral sprouts and dense vertical regeneration of axons from the deeper dermis led to complete, rapid reinnervation of the epidermis at the axotomy site. In contrast to the slower, incomplete reinnervation previously noted in humans after this technique, in both SIV infected and uninfected macaques epidermal reinnervation was rapid and completed by 56 days postaxotomy. p75 was densely expressed on the Schwann cells of uninjured nerve bundles along the excision line and on epidermal Schwann cell processes. In both SIV-infected and uninfected macaques, Schwann cell process density was highest at the earliest timepoints postaxotomy and then declined at a similar rate. However, SIV-infection delayed epidermal nerve fiber regeneration and remodeling of new sprouts at every timepoint postaxotomy, and SIV-infected animals consistently had lower mean epidermal Schwann cell densities, suggesting that Schwann cell guidance and support of epidermal nerve fiber regeneration may account for altered nerve regeneration. The relatively rapid regeneration time and the completeness of epidermal reinnervation in this macaque model provides a useful platform for assessing the efficacy of neurotrophic or regenerative drugs for sensory neuropathies including those caused by HIV, diabetes mellitus, medications, and toxins. PMID- 19296478 TI - Evaluation of indole-based probes for high-throughput screening of drug binding to human serum albumin: Analysis by high-performance affinity chromatography. AB - There has been growing interest in the use of rapid and selective separation methods such as high-performance affinity chromatography (HPAC) or affinity capillary electrophoresis (ACE) for the characterization of drug-protein interactions. L-Tryptophan is commonly used in these and other methods as a site selective probe for examining the binding of small solutes and drugs at Sudlow site II on the protein HSA. However, solutions of L-tryptophan can be unstable and are generally prepared fresh daily for these studies. In this report, HPAC was used to examine other indole compounds as possible replacements for L tryptophan as a site-selective probe for use in the high-throughput screening of drug binding to HSA; the implications of these results in the use of such compounds in ACE were also considered. The probe candidates that were tested included indole-3-acetic acid, indole-3-carboxylic acid, indole-3-butyric acid, indole-3-propionic acid, indole-3-methanol, 3-acetylindole, and 3-methylindole. All of these compounds were found by (1)H NMR and UV-Vis spectroscopy to be stable for up to 3 wk at room temperature when kept in a pH 7.4, 0.067 M phosphate buffer. The binding of these compounds was examined by using columns that contained immobilized HSA. 3-Acetylindole was found to be the best candidate in this group for use as an alternative probe to L-tryptophan for Sudlow site II. This probe had the same binding site and a similar affinity to L-tryptophan but was more stable in aqueous solution, making it suitable for high-throughput screening of drug-HSA binding in both HPAC and ACE. PMID- 19296479 TI - Characterization of high molecular weight multimeric states of human haptoglobin and hemoglobin-based oxygen carriers by high-mass MALDI MS. AB - High-mass MALDI-TOF mass spectrometry (MS) is a novel analytical approach to study large biomolecules and their interactions. It is a powerful alternative method to gel electrophoresis (GE) and size exclusion chromatography (SEC) for obtaining information on the molecular weights of macromolecules and for determining protein complexes. The precision of mass measurements (mass accuracy), high sensitivity, speed of the analysis, and tolerance toward sample heterogeneity are the major features of this MS-based approach. Remarkably, MS provides direct stoichiometric information of macromolecular protein complexes, when noncovalent interactions are stabilized during desorption/ionization by use of chemical cross-linking reagents. In this study, high-mass MALDI-TOF MS was applied to characterize the multimeric state of the human plasma protein haptoglobin (Hp), which is in the mass range of 150-300 kDa. Also, higher order structures of hemoglobin-based oxygen carriers (HBOCs) and their interactions with human haptoglobin were analyzed. These investigations are of clinical importance and contribute to the overall understanding of specific toxicity and clearance of HBOCs. PMID- 19296480 TI - De novo expression of the neurokinin 1 receptor in spinal lamina I pyramidal neurons in polyarthritis. AB - Spinal lamina I (LI) neurons play a major role in the transmission and integration of pain-related information that is relayed to higher centers. Alterations in the excitability of these neurons influence chronic pain development, and expression of the neurokinin 1 receptor (NK-1r) is thought to play a major role in such changes. Novel expression of NK-1r may underlie hyperexcitability in new populations of LI neurons. LI projection neurons can be classified morphologically into fusiform, pyramidal, and multipolar cells, differing in their functional properties, with the pyramidal type being nonnociceptive. In agreement with this, we have shown that spinoparabrachial pyramidal neurons seldom express NK-1r, in contrast with the other two cell types. In this study we investigated in the rat the long-term changes in NK-1r expression by spinoparabrachial LI neurons following the unilateral injection in the hindpaw plantar surface of complete Freund's adjuvant (CFA). Cholera toxin subunit B (CTb) was injected unilaterally into the parabrachial nucleus. Our results revealed that, ipsilaterally, pyramidal neurons were seldom immunoreactive for NK-1r both in saline-injected and in CFA-injected rats, up to 10 days post-CFA. However, a considerable number of pyramidal cells were immunoreactive for NK-1r at 15, 21, and 30 days post-CFA. Our data raise the possibility -- which needs to be confirmed by electrophysiology -- that most LI projection neurons of the pyramidal type are likely nonnociceptive in naive animals but might become nociceptive following the development of arthritis. PMID- 19296481 TI - Pyrrolopyrrole cyanine dyes: a new class of near-infrared dyes and fluorophores. AB - Pyrrolopyrrole cyanine (PPCy) dyes are presented as a novel class of near infrared (NIR) chromophores, which are synthesized in a condensation reaction of diketopyrrolopyrrole with heteroarylacetonitrile compounds. Their optical properties are marked by strong and narrow-band NIR absorptions. Complexation products with BF(2) and BPh(2) show strong NIR fluorescence and hardly any absorption in the visible range. We synthesized a series of new PPCys that differ only in the heterocyclic peripheral groups of the chromophore. With this strategy, the absorption spectra can be tuned between 684 and 864 nm, while high fluorescence quantum yields are maintained. The influence of the heterocycle on the optical properties of the dyes is discussed. PMID- 19296482 TI - Iron salts in the catalyzed synthesis of 5-substituted 1H-tetrazoles. AB - Clicking iron: Cheap and environmentally friendly [Fe(OAc)(2)] is used for the catalysis of cycloadditions between aryl nitriles and trimethylsilyl azide to prepare substituted 1H-tetrazoles in good yield (see scheme). PMID- 19296483 TI - Controllable red, green, blue (RGB) and bright white upconversion luminescence of Lu2O3:Yb3+/Er3+/Tm3+ nanocrystals through single laser excitation at 980 nm. AB - Light fantastic! Lu(2)O(3):Yb(3+)/Er(3+)/Tm(3+) nanocrystals with controllable red, green, blue (RGB) and bright white upconversion luminescence by a single laser excitation of 980 nm have been successfully synthesized (see picture). Due to abundant UC PL colors, it can potentially be used as fluorophores in the field of color displays, back light, UC lasers, photonics, and biomedicine.Lu(2)O(3):Yb(3+)/Er(3+)/Tm(3+) nanocrystals have been successfully synthesized by a solvothermal process followed by a subsequent heat treatment at 800 degrees C. Powder X-ray diffraction, transmission electron microscopy, upconversion photoluminescence spectra, and kinetic decay were used to characterize the samples. Under single-wavelength diode laser excitation of 980 nm, the bright blue emissions of Lu(2)O(3):Yb(3+), Tm(3+) nanocrystals near 477 and 490 nm were observed due to the (1)G(4)-->(3)H(6) transition of Tm(3+). The bright green UC emissions of Lu(2)O(3):Er(3+) nanocrystals appeared near 540 and 565 nm were observed and assigned to the (2)H(11/2)-->(4)I(15/2) and (4)S(3/2)- >(4)I(15/2) transitions, respectively, of Er(3+). The ratio of the intensity of green luminescence to that of red luminescence decreases with an increase of concentration of Yb(3+) in Lu(2)O(3):Er(3+) nanocrystals. In sufficient quantities of Yb(3+) with resprct to Er(3+), the bright red UC emission of Lu(2)O(3):Yb(3+)/Er(3+) centered at 662 nm was predominant, due to the (4)F(9/2)- >(4)I(15/2) transition of Er(3+). Based on the generation of red, green, and blue emissions in the different doped Lu(2)O(3):RE(3+) nanocrystals, it is possible to produce the luminescence with a wide spectrum of colors, including white, by the appropriate doping of Yb(3+), Tm(3+), and Er(3+) in the present Lu(2)O(3) nanocrystals. Namely, Lu(2)O(3):3 %Yb(3+)/0.2 %Tm(3+)/0.4 %Er(3+) nanocrystals show suitable intensities of blue, green, and red (RGB) emission, resulting in the production of perfect and bright white light with CIE-x=0.3456 and CIE y=0.3179, which is very close to the standard equal energy white light illuminate (x=0.33, y=0.33). Because of abundant luminescent colors from RGB to white in Lu(2)O(3):Yb(3+)/Er(3+)/Tm(3+) nanocrystals under 980 nm laser diode (LD) excitation, they can potentially be used as fluorophores in the field of color displays, back light, UC lasers, photonics, and biomedicine. PMID- 19296484 TI - Growth mechanism and chemical bonding in scandium-doped copper clusters: experimental and theoretical study in concert. AB - Size matters! The electronic structure and size-dependent stability of neutral and cationic scandium-doped copper clusters have been investigated by mass spectrometric studies (for the cations) and also quantum chemical computations. The proposed reaction paths ultimately lead to the most stable Frank-Kasper shaped Cu(16)Sc(+) cluster (shown here), which could be the germ of a new crystallization process.Electronic structure and size-dependent stability of scandium-doped copper cluster cations, Cu(n)Sc(+), were investigated by using a dual-target dual-laser vaporization production scheme followed by mass spectrometric studies and also quantum chemical computations in the density functional theory framework. The neutral species also were studied by using computational methods. Enhanced abundances and dissociation energies were measured in the case of Cu(n)Sc(+) for n=4, 6, 8, 10 and 16, the last of these identified as being extraordinary stable. Neutral clusters are stable with n=5, 7, 9 and 15, which are isoelectronic with respect to the number of the valence s electrons with the stable cationic clusters; hence a simple electron count determines cluster properties to a great extent. The Cu(17)Sc cluster was found to be a superatomic molecule, containing Cu(16)Sc(+) and Cu(-) units; however, the charge separation is not as pronounced as in the case of CuLi. Cu(15)Sc was found to be a stable cluster with a large dissociation energy and a closed electronic structure; hence this can be regarded as a superatom, analogous to the noble gases. The main factors determining the growth patterns of these clusters are the central position of the scandium atom and the successive filling of the shell orbitals. For smaller clusters, the reaction paths appear to diverge yielding various products; however all paths ultimately lead to the most stable Frank-Kasper shaped Cu(16)Sc cluster, which in turn can be the germ of the crystallization process. PMID- 19296485 TI - Strong adsorbability of mercury ions on aniline/sulfoanisidine copolymer nanosorbents. AB - The highest Hg-ion adsorbance so far, namely up to 2063 mg g(-1), has been achieved by poly(aniline-co-5-sulfo-2-anisidine) nanosorbents. Sorption of Hg ions occurs mainly by redox and chelation mechanisms (see scheme), but also by ion exchange and physisorption.Poly(aniline (AN)-co-5-sulfo-2-anisidine (SA)) nanoparticles were synthesized by chemical oxidative copolymerization of AN and SA monomers, and their extremely strong adsorption of mercury ions in aqueous solution was demonstrated. The reactivity ratios of AN and SA comonomers were found to be 2.05 and 0.02, respectively. While AN monomer tends to homopolymerize, SA monomer tends to copolymerize with AN monomer because of the great steric hindrance and electron-attracting effect of the sulfo groups, despite the effect of conjugation of the methoxyl group with the benzene ring. The effects of initial mercury(II) concentration, sorption time, sorption temperature, ultrasonic treatment, and sorbent dosage on mercury-ion sorption onto AN/SA (50/50) copolymer nanoparticles with a number-average diameter of around 120 nm were significantly optimized. The results show that the maximum Hg ion sorption capacity on the particulate nanosorbents can even reach 2063 mg of Hg per gram of sorbent, which would be the highest Hg-ion adsorbance so far. The sorption data fit to the Langmuir isotherm, and the process obeys pseudo-second order kinetics. The IR and UV/Vis spectral data of the Hg-loaded copolymer particles suggest that some mercury(II) was directly reduced by the copolymer to mercury(I) and even mercury(0). A mechanism of sorption between the particles and Hg ions in aqueous solution is proposed, and a physical/ion exchange/chelation/redox sorption ratio of around 2/3/45/50 was found. Copolymer nanoparticles may be one of the most powerful and cost-effective sorbents of mercury ions, with a wide range of potential applications for the efficient removal and even recovery of the mercury ions from aqueous solution. PMID- 19296486 TI - Macrocyclic hexaureas: synthesis, conformation, and anion binding. AB - Five macrocylic compounds XXXXXX, XXDXXD, XDXDXD, XDDXDD, and DDDDDD with 48 membered rings, in which six xanthene and/or diphenyl ether fragments are linked through six urea (-NH-C(O)-NH-) groups, have been synthesized. In the cyclization step, a linear diamine was allowed to react with the appropriate diisocyanate by using a [5+1] (i.e., "XDXDX+D" for XDXDXD), [4+2] (DDDDDD), or [3+3] (XDDXDD) procedure. Compounds XXXXXX and XXDXXD were prepared from two molecules of the dimeric amine XX and two molecules of the respective monomeric diisocyanate (X or D) in a [2+1+2+1] (or 2x[2+1]) reaction. The (nonoptimized) yields in the cyclization step ranged from 45 to 80%. The linear precursor diamines or diisocyanates were obtained by analogous condensation reactions by using partial protection with a tert-butoxycarbonyl group. All the macrocyclic compounds and synthetic intermediates were characterized by (1)H NMR and mass spectra. Three different crystal structures were obtained for XDDXDD, which show the molecule in a more or less strongly folded conformation determined by intramolecular hydrogen bonding. The interaction of the hexaureas with selected anions was studied by (1)H NMR spectroscopy and UV absorption spectrophotometry. PMID- 19296487 TI - Actin isoform expression patterns during mammalian development and in pathology: insights from mouse models. AB - The dynamic actin cytoskeleton, consisting of six actin isoforms in mammals and a variety of actin binding proteins is essential for all developmental processes and for the viability of the adult organism. Actin isoform specific functions have been proposed for muscle contraction, cell migration, endo- and exocytosis and maintaining cell shape. However, these specific functions for each of the actin isoforms during development are not well understood. Based on transgenic mouse models, we will discuss the expression patterns of the six conventional actin isoforms in mammals during development and adult life. Ablation of actin genes usually leads to lethality and affects expression of other actin isoforms at the cell or tissue level. A good knowledge of their expression and functions will contribute to fully understand severe phenotypes or diseases caused by mutations in actin isoforms. PMID- 19296488 TI - Cytoskeleton rearrangements during Listeria infection: clathrin and septins as new players in the game. AB - The study of an infection process can reveal how microbes exploit the host, and can illuminate unknown host cellular functions. Invasive pathogens have evolved efficient strategies to promote their internalization within normally non phagocytic host cells. The so-called "zippering" bacteria present to host cell receptors molecules that mimic endogenous ligands, thereby inducing specific intracellular signaling cascades ultimately resulting in actin polymerization and uptake. Here we review how the bacterial pathogen Listeria monocytogenes enters into cells, and present a series of studies revealing that in addition to actin rearrangements this bacterium exploits the clathrin-mediated endocytosis machinery together with septins, a novel cytoskeleton element. The challenge is now to decipher how all of these components orchestrate themselves to permit entry into normally non-phagocytic cells. PMID- 19296489 TI - Detection of fruit by the Cerrado's marmoset (Callithrix penicillata): modeling color signals for different background scenarios and ambient light intensities. AB - Among placental mammals, only primates have trichromatic color vision, however this is not a uniform condition. Under different genetic status, Old World monkeys have routine trichromacy, while New World monkeys show a visual polymorphism, characterized by obligatory male dichromacy. The ecological role of this genetic difference still remains unclear, but some studies show that dichromats and trichromats appear to have different abilities in detecting colored targets against a background of leaves. The Cerrado's marmoset (Callithrix penicillata) is known to forage in brightly illuminated (savanna-like vegetation) and dimly illuminated (forests) environments, exploiting a high amount of dark fruits. Hence, it seems to be a good model for studying the differential advantages enjoyed by each color vision phenotype under natural conditions. Our aim was to verify how the different phenotypes of Cerrado's marmoset detect components of their diet, evaluating the existence of differential phenotype advantages. Under two different light conditions, visual signals of naturally consumed fruits were modeled against different backgrounds scenarios. Even though dichromats and trichromats appear to be equally suited for tasks involving fruit detection, phenotype differential advantages are observed in this marmoset. In many conditions trichromats are predicted to perform better than dichromats, but under low ambient light dichromats manage to outperform trichromats in some scenarios. Phenotypes that carry widely spaced and longer M/L pigments enjoy the most advantage. These differential performances of trichromatic phenotypes, together with overdominance selection, seem to explain the maintenance of the tri-allelic system found in callitrichids. PMID- 19296490 TI - Changes in muscle force-length properties affect the early rise of force in vivo. AB - Changes in contractile rate of force development (RFD), measured within a short time interval from contraction initiation, were measured after a period of strength training that led to increases in muscle fascicle length but no measurable change in neuromuscular activity. The relationship between training induced shifts in the moment-angle relation and changes in RFD measured to 30 ms (i.e., early) and 200 ms (i.e., late) from the onset of isometric knee extension force were examined; shifts in the moment-angle relation were used as an overall measure of changes in quadriceps muscle fascicle length. A significant proportion of the variance in RFD measured only in the initial contraction phase (0-30 ms) could be explained by shifts in the moment-angle relation (r=-0.66-0.71; R2=0.44 0.50). Training-induced increases in muscle fascicle length may lead to a reduced or complete lack of adaptive gains in contractile RFD, especially in the early contraction phase. PMID- 19296491 TI - Long-term dietary administration of valproic acid does not affect, while retinoic acid decreases, the lifespan of G93A mice, a model for amyotrophic lateral sclerosis. AB - Mice bearing the mutated gene for Cu/Zn superoxide dismutase (G93A) are a good model for human amyotrophic lateral sclerosis (ALS). They develop progressive limb paralysis paralleled by loss of motor neurons of the cervical and lumbar spinal cord, which starts at 3-3.5 months of age and ends with death at 4-5 months. Several treatments have been attempted to delay clinical symptoms and to extend lifespan, and some have had modest beneficial effects. One such treatment, based on long-term administration of valproic acid (VPA), resulted in controversial results. We report here that, while dietary supplementation with high VPA dosage slows down motor neuron death, as assessed by measurement of a specific marker for cholinergic neurons in the spinal cord, it has no significant effect on lifespan. Recently, the hypothesis has been put forward that a deficiency of retinoic acid (RA) and its signaling may have a role in ALS. We report that long-term dietary supplementation with RA has no effect on the decrease of the cholinergic marker in the spinal cord, but it significantly shortens lifespan of G93A mice. PMID- 19296494 TI - Eotaxin-1, -2, and -3 immunoreactivity and protein concentration in the nasal polyps of eosinophilic chronic rhinosinusitis patients. AB - OBJECTIVES/HYPOTHESIS: Eosinophilic chronic rhinosinusitis (CRS) is characterized by the accumulation of numerous eosinophils in the sinus mucosa and nasal polyps, which are frequently difficult to control, even with surgery. The present study was designed to evaluate the expression and localization of eotaxins, which are well known to be potent and selective chemoattractants for eosinophils in CRS. STUDY DESIGN: Randomized study. METHODS: The patients were classified into eosinophilic and noneosinophilic groups. Histopathological profiles of the nasal polyp were observed with hematoxylin-eosin staining. Eotaxin-1, -2, and -3 were immunohistochemically stained in the nasal polyps. Furthermore, the protein content of eotaxin subtypes inside the nasal polyp and sinus effusion was measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: In the nasal polyps, immunoreactivities of the eotaxin subfamily, eotaxin-1, -2, and -3, were noted in most of the infiltrating eosinophils, as well as in other inflammatory cells, epithelial cells, and endothelial cells. Compared with noneosinophilic CRS groups, eosinophilic CRS groups had a significant expression of eotaxins in their eosinophils. The eotaxin concentrations of nasal polyp and sinus effusion as measured by ELISA were significantly increased in the eosinophilic CRS group compared to the noneosinophilic CRS group. CONCLUSIONS: The present findings suggest that enhanced eotaxin family production by eosinophils results in the recruitment of eosinophils into the tissue by a self-amplifying process. Laryngoscope, 2009. PMID- 19296495 TI - Effects of systemic transplantation of adipose tissue-derived stem cells on olfactory epithelium regeneration. AB - OBJECTIVE/HYPOTHESIS: The purpose of the study was to investigate the effect of intravenous adipose tissue-derived stem cell (ADSC) transplantation on olfactory epithelium regeneration following transection of the olfactory nerve in rats. STUDY DESIGN: This was a experimental study using primary cultures of mesenchymal stem cells derived from animal adipose tissue with histological analysis of animal olfactory tissue. METHODS: All rats underwent unilateral transection of the olfactory nerve to induce degeneration of olfactory epithelium, and then were observed for regeneration according to time sequences. ADSCs were cultivated from neck adipose tissue of rats, and systemically injected into the experimental group. The control group was injected with phosphate buffered solution, instead of ADSCs. After 30 days, regeneration of olfactory epithelium was observed with olfactory marker protein (OMP) and proliferating cell nuclear antigen. To observe the characteristics of the transplanted ADSCs, olfactory epithelium was stained with von Willebrant factor and OMP. RESULTS: After olfactory nerve transection, mature olfactory cells disappeared in 5 days, but gradually regained their thickness with increased cell numbers at approximately 10 to 15 days. By 30 days post-transection, the thickness and cellular composition of epithelium was almost restored to baseline levels pretransection. However, OMP expressions remained decreased compared with day 0 or 3. Systemically injected ADSCs were transplanted into the olfactory epithelium and survived beyond 4 weeks. The ADSCs promoted regeneration of olfactory epithelium in the animal model and differentiated into olfactory receptor neurons and endothelial cells. CONCLUSIONS: Our findings suggest the feasibility of ADSC transplantation as a treatment for head trauma related olfactory dysfunction. PMID- 19296496 TI - Comparison of transnasal endoscopic and open craniofacial resection for malignant tumors of the anterior skull base. AB - OBJECTIVE/HYPOTHESIS: Craniofacial resection (CFR) represents the traditional approach for resection of anterior skull base (ASB) malignancies. However, this past decade has witnessed the emergence of transnasal endoscopic ASB resection (TER) as a feasible alternative. The aim of this study was to compare TER and CFR for ASB malignancy resection. METHODS: Retrospective analysis at a tertiary care medical center on 66 patients undergoing ASB malignant tumor resection between September 1997 and December 2006. RESULTS: Eighteen patients were treated with TER, whereas 48 patients underwent CFR. The two groups showed no significant difference in complication rate (27.8% for TER and 25.0% for CFR, P > 0.05), survival (94.4% for TER and 83.3% for CFR, P > .05), and metastasis (11.1% for TER and 8.3% for CFR, P > .05). A significant difference was observed in hospital stay with an average of 3.8 days in the TER group compared to 8.1 days in the CFR group (P < .05). Local recurrence approximated significance (5.6% for TER and 29.2% for CFR, P = .051) CONCLUSIONS: TER of ASB malignancy is associated with a decreased hospital stay and faster recovery when compared to open CFR. Lower local recurrence rate in the TER group may reflect a discrepancy in histology and clinical stage. We found no significant differences in survival, metastatic, or complication rates in the two groups, whereas patients in the TER group had the added benefit of a desirable cosmetic outcome. Overall, TER seems to be an excellent alternative to CFR in properly selected cases. PMID- 19296497 TI - Head and neck reconstruction with the deep inferior epigastric perforator flap: a report of two cases. AB - Rectus abdominis musculocutaneous (RAMC) free flaps are preferred for head and neck reconstruction because of the abundant blood supply to the rectus abdominis musculocutaneous. In contrast, the indications for deep inferior epigastric perforator (DIEP) free flaps in head and neck reconstruction are limited. In this report, two cases of oral cavity reconstruction with DIEP free flaps are described. In both cases, the defect was reconstructed with a DIEP free flap because it could avoid functional damage to the donor site. Successful reconstruction with a two skin-island method was performed in both patients. Furthermore, donor site morbidity was minimal in both patients. When a DIEP free flap is used for head and neck reconstruction, elimination of dead space is the most difficult problem, because a DIEP free flap does not contain well vascularized muscle tissue. We compensate for this disadvantage with a flap designed to include a de-epithelialized skin flap. Although this technique is not always the first choice for head and neck reconstruction, it is suitable for patients who wish to avoid donor site morbidity. PMID- 19296498 TI - Versatility of the vertical designed deep inferior epigastric perforator flap. AB - The deep inferior epigastric perforator (DIEP) flap is one of the most commonly used perforator flaps. However, when horizontally oriented, it has certain pitfalls such as sacrifice of the contralateral donor site, the need to discard soft tissue that will not be adequately perfused, and long operating times. To overcome these handicaps, the flap in a vertical orientation was designed and demonstrated the versatility of this technique in a number of different anatomical tissue restorations in this study. The vertical DIEP flaps were used in 8 patients (7 males and 1 female) in 3 years. Defects were located in the lower limb (4 cases), face (3 cases), and hip (1 cases). All flaps were transferred as free, except one in the hip that was pedicled. Flap sizes varied from 7 x 5 cm to 13 x 9 cm. All donor defects were closed primarily. The mean follow-up time was 13.6 months. All flaps survived totally, but one which had partial necrosis. A hematoma developed postoperatively in the donor site of the pedicled DIEP flap. Cosmetic outcomes were satisfactory in all patients. All donor site scars were acceptable with minimal contour irregularities. The vertical DIEP flap offers diverse opportunities in various soft tissue reconstructions and is candidate to be an important choice of free flap repertoire. The method can be proposed particularly for restoration of moderate sized defects rather than that of large defects as in postmastectomy breast reconstruction. PMID- 19296499 TI - Sural perforator flap: assessment of the posterior calf region as donor site for a free fasciocutaneous flap. AB - Three kinds of free fasciocutaneous flap from the posterior calf region have been described in the literature: the medial sural perforator flap, the lateral sural perforator flap, and the traditional posterior calf fasciocutaneous flap that is supplied by superficial cutaneous vessels. Moreover, it has been reported that superficial cutaneous vessels are of a suitable size for microanastomosis when deep musculocutaneous perforators are absent or relatively tiny. To establish a safe technique for free fasciocutaneous flap elevation from the posterior calf region, we examined the number and location of the musculocutaneous perforators and the size of superficial cutaneous vessels at their origin from the popliteal artery in six formalinized cadavers. We found that all legs had at least one perforator either from the medial sural artery or the lateral sural artery. By contrast, we failed to find superficial cutaneous vessels of suitable size for microanastomosis in three legs, and there was no significant inverse relationship between the diameter of the superficial cutaneous artery and the number of musculocutaneous perforators. Our results suggest that the medial sural perforator flap and the lateral sural perforator flap might be the surgeon's first and second choice, respectively. The traditional posterior calf fasciocutaneous flap should be the third choice because our study suggests that its availability is doubtful. Another site is recommended, when preoperative Doppler study suggests that the existence of musculocutaneous perforator is in doubt. Two clinical cases, with a medial sural perforator flap and a lateral sural perforator flap, respectively, are presented. PMID- 19296500 TI - Nonparallel cutaneous microcirculatory responses to pharmacologic alterations of systemic arterial pressure in rats. AB - Both hypotension and hypertension aggravate the damage of reperfusion injury after reconstructive microsurgery. The purpose of this study is to establish a theoretical guide for postoperative blood pressure control in optimizing the cutaneous perfusion and flap survival. Systemic arterial pressure was altered by the intravenous infusion of saline, sodium nitroprusside, phentolamine, and phenylephrine in thirty-two 280-350 g anesthetized Sprague Dawley rats. Power spectral analysis of systemic arterial pressure (SAP) and laser Doppler flowmetry (flux) of epigastric skin were used to reveal the blood pressure and cutaneous blood flow variabilities. Nonparallel responses of cutaneous perfusion and blood pressure were found. The baseline SAP and flux were 126.0 +/- 1.4 mmHg and 57.2 +/- 1.8 au, respectively. Sodium nitroprusside and phentolamine significantly decreased the SAP (71.1 +/- 2.7 and 70.5 +/- 1.5 mmHg, P < 0.0001). However, the corresponding responses in cutaneous perfusion were opposite (56.2 +/- 3.1 au, P = 0.7389 and 36.2 +/- 2.3 au, P < 0.0001). Phenylephrine significantly increased the SAP (171.7 +/- 3.0 mmHg, P < 0.0001) but the flux of epigastric skin was decreased (44.4 +/- 2.6 au, P < 0.0001). Phentolamine and phenylephrine showed negative effects on the systemic cardiac and vascular sympathetic modulations. Sodium nitroprusside had a trend in increasing systemic vasomotor activity. We suggested not using vasoconstrictors in treating intra and postoperative hypotension associated with free flap transfer. Nitric oxide donors are superior to alpha-adrenoceptor antagonists in preserving the cutaneous flap perfusion when treating postoperative hypertension. PMID- 19296501 TI - Technique and proof of patency of microsurgical lympho-lymphonodular anastomoses: a study in the rat model. AB - BACKGROUND: Lymphedemas due to local lymphatic blocks can be treated by microsurgical transplantation or transposition of lymphatic vessels. Here, the anastomoses are usually made end-to-end between lymphatics, but occasionally appropriate lymphatic recipient vessels are missing. In such cases, reconstructing lymph drainage by connection to a lymph node could be another technical option. The purpose of this study was to examine the patency rate of such lympho-lymphonodular anastomoses in an experimental animal model. METHODS: Male Sprague-Dawley rats were anesthetized, and the retroperitoneum was exposed. Patent blue dye was injected into the left foot to stain lymphatic structures. In group A (n = 8), the left lumbar trunk was cut centrally, the distal part was turned over to the right lumbar lymph node, and a microsurgical lympho lymphonodular anastomosis was performed. In group B (n = 8), the left lumbar trunk was cut. After 8 weeks, the lumbar region was surgically re-explored, and the lymphatic drainage was examined by injection of Patent blue dye into the left lumbar lymph node. RESULTS: In 8/8 animals of group A, patent transposed lymphatics were found. The patency of the anastomosis was proven directly by observation of blue dye transit and indirectly by observation of blue staining of the right lumbar lymph node. In 6/8 animals of group B, no lymphatic connection to the right lumbar lymphatic system was observed. CONCLUSIONS: This is the first report of the microsurgical technique and the proof of patency of lympho lymphonodular anastomoses. The novel animal model for testing the patency of transposed lymphatics is discussed. PMID- 19296502 TI - Improvement of peripheral nerve regeneration in acellular nerve grafts with local release of nerve growth factor. AB - Previous studies have demonstrated the potential of growth factors in peripheral nerve regeneration. A method was developed for sustained delivery of nerve growth factor (NGF) for nerve repair with acellular nerve grafts to augment peripheral nerve regeneration. NGF-containing polymeric microspheres were fixed with fibrin glue around chemically extracted acellular nerve grafts for prolonged, site specific delivery of NGF. A total of 52 Wistar rats were randomly divided into four groups for treatment: autografting, NGF-treated acellular grafting, acellular grafting alone, and acellular grafting with fibrin glue. The model of a 10-mm sciatic nerve with a 10-mm gap was used to assess nerve regeneration. At the 2nd week after nerve repair, the length of axonal regeneration was longer with NGF-treated acellular grafting than acellular grafting alone and acellular grafting with fibrin glue, but shorter than autografting (P < 0.05). Sixteen weeks after nerve repair, nerve regeneration was assessed functionally and histomorphometrically. The percentage tension of the triceps surae muscles in the autograft group was 85.33 +/- 5.59%, significantly higher than that of NGF treated group, acellular graft group and fibrin-glue group, at 69.79 +/- 5.31%, 64.46 +/- 8.48%, and 63.35 +/- 6.40%, respectively (P < 0.05). The ratio of conserved muscle-mass was greater in the NGF-treated group (53.73 +/- 4.56%) than in the acellular graft (46.37 +/- 5.68%) and fibrin glue groups (45.78 +/- 7.14%) but lower than in the autograft group (62.54 +/- 8.25%) (P < 0.05). Image analysis on histological observation revealed axonal diameter, axon number, and myelin thickness better with NGF-treated acellular grafting than with acellular grafting alone and acellular grafting with fibrin glue (P < 0.05). There were no significant differences between NGF-treated acellular grafting and autografting. This method of sustained site-specific delivery of NGF can enhance peripheral nerve regeneration across short nerve gaps repaired with acellular nerve grafts. PMID- 19296503 TI - Lean and obese Zucker rats exhibit different patterns of p70s6 kinase regulation in the tibialis anterior muscle in response to high-force muscle contraction. AB - Increased phosphorylation of the 70-kDa ribosomal S6 kinase (p70S6k) signaling is strongly correlated with the degree of muscle adaptation following exercise. Herein we compare the phosphorylation of p70S6k, Akt, and mammalian target of rapamycin (mTOR) in the tibialis anterior (TA) muscles of lean and obese Zucker rats following a bout of eccentric exercise. Exercise increased p70S6k (Thr389) phosphorylation immediately after (33.3+/-7.2%) and during [1 h (24.0+/-14.9%) and 3 h (24.6+/-11.3%)] recovery in the lean TA and at 3 h (33.5+/-8.0%) in the obese TA Zucker rats. mTOR (Ser2448) phosphorylation was elevated in the lean TA immediately after exercise (96.5+/-40.3%) but remained unaltered in the obese TA. Exercise increased Akt (Thr308) and Akt (Ser473) phosphorylation in the lean but not the obese TA. These results suggest that insulin resistance is associated with alterations in the ability of muscle to activate p70S6k signaling following an acute bout of exercise. PMID- 19296504 TI - Cerebrospinal fluid biomarker signature in Alzheimer's disease neuroimaging initiative subjects. AB - OBJECTIVE: Develop a cerebrospinal fluid biomarker signature for mild Alzheimer's disease (AD) in Alzheimer's Disease Neuroimaging Initiative (ADNI) subjects. METHODS: Amyloid-beta 1 to 42 peptide (A beta(1-42)), total tau (t-tau), and tau phosphorylated at the threonine 181 were measured in (1) cerebrospinal fluid (CSF) samples obtained during baseline evaluation of 100 mild AD, 196 mild cognitive impairment, and 114 elderly cognitively normal (NC) subjects in ADNI; and (2) independent 56 autopsy-confirmed AD cases and 52 age-matched elderly NCs using a multiplex immunoassay. Detection of an AD CSF profile for t-tau and A beta(1-42) in ADNI subjects was achieved using receiver operating characteristic cut points and logistic regression models derived from the autopsy-confirmed CSF data. RESULTS: CSF A beta(1-42) was the most sensitive biomarker for AD in the autopsy cohort of CSF samples: receiver operating characteristic area under the curve of 0.913 and sensitivity for AD detection of 96.4%. In the ADNI cohort, a logistic regression model for A beta(1-42), t-tau, and APO epsilon 4 allele count provided the best assessment delineation of mild AD. An AD-like baseline CSF profile for t-tau/A beta(1-42) was detected in 33 of 37 ADNI mild cognitive impairment subjects who converted to probable AD during the first year of the study. INTERPRETATION: The CSF biomarker signature of AD defined by A beta(1-42) and t-tau in the autopsy-confirmed AD cohort and confirmed in the cohort followed in ADNI for 12 months detects mild AD in a large, multisite, prospective clinical investigation, and this signature appears to predict conversion from mild cognitive impairment to AD. PMID- 19296505 TI - Supracricoid laryngectomy: age influence on long-term functional results. AB - OBJECTIVES/HYPOTHESIS: : Supracricoid laryngectomies (SCL) are conservative surgical techniques for the treatment of selected laryngeal carcinomas, currently adopted also in old patients. Long-term functional results have not been reported in elderly patients. The aim of this study is to compare voice and swallowing in elderly and younger patients following SCL. STUDY DESIGN: : Cross-sectional study. METHODS: : Twenty male patients who underwent SCL were recruited in the study; 10 were younger than 65 years at the time of surgery and 10 were older. Video-endoscopic ratings were taken of neoglottic vibration patterns and bolus transit. Voices were perceptually rated using the GIRBAS scale. The maximum phonation time (MPT) and the syllables diadochokinesis were measured. Spectrograms were recorded. All of the patients completed a self-assessment questionnaire for both voice and swallowing. The data obtained from the two groups were compared through the Mann-Whitney test. RESULTS: : Video-endoscopic ratings of neoglottal vibration and bolus transit showed no difference between the two groups. The perceptual assessment showed a harsh voice in both groups. No significant difference was found for mean syllable diadochokinesis and the mean MPT was 6.3 seconds and 8.8 seconds, respectively in the younger and older group. The mean value of the Yanagihara scale of voice spectrogram was 3.8 and 3.7. Voice and swallowing quality-of-life questionnaires revealed satisfied patients in both age groups. CONCLUSIONS: : Age by itself does not have a significant impact on long-term functional results following SCL. Meticulous selection of the candidate to SCL allows the application of this surgical technique with adequate long-term functional results. Laryngoscope, 2009. PMID- 19296506 TI - Efficacy of neck dissection: are surgical volumes important? AB - OBJECTIVES/HYPHOTHESIS: Surgical volumes have been shown in many fields to reflect expertise and to be associated with improved clinical outcomes. We sought to test similar hypotheses that neck dissection is a volume-sensitive procedure wherein the number of lymph nodes harvested and clinical outcomes are correlated with the number of neck dissections performed. METHODS: We conducted a retrospective analysis of 375 neck dissections. The number of nodes harvested was examined by multiple linear regression before and after mathematical correction for pathologists' variation over time. The prior experiences of the surgeons were also averaged, depending on whether there was regional recurrence after at least 9 months of follow-up. RESULTS: More nodes were found by the pathologists over time, with 2.1 more nodes being found on average for each successive year (P < .001). After correcting for this effect of time, there remained a significant relationship between surgeons' experience and number of nodes harvested (P < .003). Additionally, cases that recurred were operated by less-experienced surgeons (P = .02). CONCLUSIONS: We have demonstrated that there is a "learning curve" for neck dissection. Our most experienced surgeon harvested on average 11 more nodes than similar specimens submitted by the group of inexperienced surgeons. The effect of experience remains significant even without adjusting for time (P < .001). Surgical experience also affects clinical outcome months after the dissection. This retrospective review supports surgical volume as an indicator of expertise in neck dissection that could be used to assess trainees' progress and for quality assurance in large head and neck units. Laryngoscope, 2009. PMID- 19296507 TI - Muscle senescence in short-lived wild mammals, the soricine shrews Blarina brevicauda and Sorex palustris. AB - Red-toothed (soricine) shrews are consummate predators exhibiting the highest energy turnovers and shortest life spans (ca. 18 months) of any mammal, yet virtually nothing is known regarding their physiological aging. We assessed the emerging pattern of skeletal muscle senescence (contractile/connective tissue components) in sympatric species, the semi-aquatic water shrew (WS), Sorex palustris, and the terrestrial short-tailed shrew (STS), Blarina brevicauda, to determine if muscle aging occurs in wild, short-lived mammals (H(0): shrews do not survive to an age where senescence occurs), and if so, whether these alterations are species-specific. Gracilis muscles were collected from first-year (n=17) and second-year (n=17) field-caught shrews. Consistent with typical mammalian aging, collagen content (% area) increased with age in both species (S. palustris: approximately 50%; B. brevicauda: approximately 60%). Muscle was dominated by stiffer Type I collagen, and the ratio of collagen Type I:Type III more than doubled with age. The area ratio of muscle:collagen decreased with age in both species, but was considerably lower in adult STS, suggesting species specificity of senescence. Extracellular space was age-elevated in B. brevicauda, but was preserved in S. palustris ( approximately 50 vs. 10% elevation). Though juvenile interspecific comparisons revealed no significance, adult WS myocytes had 68% larger cross-sectional area and occurred at 28% lower fibers/area than those of adult STS. We demonstrate that age-related muscle senescence does occur in wild-caught, short-lived mammals, and we therefore reject this classic aging theory tenet. Our findings moreover illustrate that differential age adjustments in contractile/connective tissue components of muscle occur in the two species of wild-caught shrews. PMID- 19296508 TI - Free-energy profiles for ions in the influenza M2-TMD channel. AB - M(2) transmembrane domain channel (M(2)-TMD) permeation properties are studied using molecular dynamics simulations of M(2)-TMD (1NYJ) embedded in a lipid bilayer (DMPC) with 1 mol/kg NaCl or KCl saline solution. This study allows examination of spontaneous cation and anion entry into the selectivity filter. Three titration states of the M(2)-TMD tetramer are modeled for which the four His(37) residues, forming the selectivity filter, are net uncharged, +2 charged, or +3 charged. M(2)-TMD structural properties from our simulations are compared with the properties of other models extracted from NMR and X-ray studies. During 10 ns simulations, chloride ions occasionally occupy the positively-charged selectivity filter region, and from umbrella sampling simulations, Cl(-) has a lower free-energy barrier in the selectivity-filter region than either Na(+) or NH(4) (+), and NH(4) (+) has a lower free-energy barrier than Na(+). For Na(+) and Cl(-), the free-energy barriers are less than 5 kcal/mol, suggesting that the 1NYJ conformation would probably not be exquisitely proton selective. We also point out a rotameric configuration of Trp(41) that could fully occlude the channel. PMID- 19296509 TI - Organotypic glioma spheroids for screening of experimental therapies: how many spheroids and sections are required? AB - Cancer spheroids are a valuable model for screening anticancer strategies. However, studies are published using various numbers of spheroids and sections per spheroid. Here, we establish the sample size requirements for valid screening strategies to treat glioma: how many spheroids per experimental group and how many sections per spheroid are required to detect one-third reduction in an endpoint measurement after treatment? From two glioblastoma patients, 32 untreated organotypic spheroids were cultured and sectioned entirely (14-100 sections per spheroid). The viable fraction was determined as endpoint by automated image analysis in sections and used to establish the minimally detectable difference between a treatment and reference group. Variance was considerable with a coefficient of variance of 21%. The biological variation in viability in sections of spheroids produced 97% of variance when sample size was large. Variance increased when numbers of spheroids but not numbers of sections per spheroid were reduced. A minimum of 12 spheroids per group and one section per spheroid was required for a valid comparison of a treatment group and a control group. When 10 treatment groups and one control group were compared, 16 spheroids per group were required. Thus, the statistical power depended almost entirely on the number of organotypic glioma spheroids and hardly on the number of sections per spheroid. The organotypic glioma spheroid model does not appear to be suitable for high-throughput screening of anticancer strategies, because of the relatively large number of spheroids required. It is the model of choice for low-throughput screening, because this model is far more representative for the parental tumor than any other more efficient glioma model. PMID- 19296510 TI - Going into lengths and widths, and depths--microscopic cytomics quantifying cell function and cell communication. PMID- 19296514 TI - Development and evaluation of BioScore: a biomarker panel to enhance prognostic algorithms for clear cell renal cell carcinoma. AB - BACKGROUND: The authors previously showed that increased tumor expression levels of B7-H1, survivin, and Ki-67 are independent predictors of poor outcome for patients with clear cell renal cell carcinoma (ccRCC). In the current study, they described the creation of a scoring system based on this panel of biomarkers that can be used in tandem with existing clinicopathologic features and algorithms to improve ccRCC outcome prediction. METHODS: The authors used immunohistochemistry to determine tumor expression levels of B7-H1, survivin, and Ki-67 for 634 consecutive ccRCC patients. A multivariate model verified that each biomarker was independently associated with RCC-specific death after adjusting for the remaining 2. A biomarker-based panel, termed BioScore, was generated to predict the likelihood of RCC-specific death. BioScore was tested for its ability to enhance the performance of several clinicopathologic features and algorithms. RESULTS: Patients with high BioScores were 5 times more likely to die from RCC compared with patients with low BioScores (hazard ratio, 5.03; 95% confidence interval, 3.82-6.61; P < .001). Multivariate adjustment for individual clinicopathologic features or existing prognostic algorithms failed to attenuate this positive association. Moreover, an examination of concordance indexes revealed that BioScore significantly enhanced the prognostic ability of each of the individual prognostic features or algorithms studied. CONCLUSIONS: The authors described the creation of BioScore, a biomarker-based scoring system that can be used in tandem with established prognostic algorithms to further enhance ccRCC outcome prediction. The need for external validation notwithstanding, they envision that BioScore can be readily updated as new biomarkers are identified. PMID- 19296515 TI - Mapping cancer mortality-to-incidence ratios to illustrate racial and sex disparities in a high-risk population. AB - BACKGROUND: Comparisons of incidence and mortality rates are the metrics used most commonly to define cancer-related racial disparities. In the US, and particularly in South Carolina, these largely disfavor African Americans (AAs). Computed from readily available data sources, the mortality-to-incidence rate ratio (MIR) provides a population-based indicator of survival. METHODS: South Carolina Central Cancer Registry incidence data and Vital Registry death data were used to construct MIRs. ArcGIS 9.2 mapping software was used to map cancer MIRs by sex and race for 8 Health Regions within South Carolina for all cancers combined and for breast, cervical, colorectal, lung, oral, and prostate cancers. RESULTS: Racial differences in cancer MIRs were observed for both sexes for all cancers combined and for most individual sites. The largest racial differences were observed for female breast, prostate, and oral cancers, and AAs had MIRs nearly twice those of European Americans (EAs). CONCLUSIONS: Comparing and mapping race- and sex-specific cancer MIRs provides a powerful way to observe the scope of the cancer problem. By using these methods, in the current study, AAs had much higher cancer MIRs compared with EAs for most cancer sites in nearly all regions of South Carolina. Future work must be directed at explaining and addressing the underlying differences in cancer outcomes by region and race. MIR mapping allows for pinpointing areas where future research has the greatest likelihood of identifying the causes of large, persistent, cancer-related disparities. Other regions with access to high-quality data may find it useful to compare MIRs and conduct MIR mapping. PMID- 19296516 TI - Racial differences in medical mistrust among men diagnosed with prostate cancer. AB - BACKGROUND: Mistrust of healthcare providers and systems is a significant barrier to quality healthcare. However, limited empirical data are available on perceptions of medical mistrust among individuals who are diagnosed with cancer. The objective of this study was to identify sociodemographic, clinical, and cultural determinants of mistrust among men diagnosed with prostate cancer. METHODS: The authors conducted an observational study among 196 African-American men (n = 71) and white men (n = 125) who were newly diagnosed with prostate cancer during 2003 through 2007. RESULTS: Race, education, healthcare experiences, and cultural factors had significant effects on mistrust. African American men (P = .01) and men who had fewer years of formal education (P = .001) reported significantly greater levels of mistrust compared with white men and men who had more education. Mistrust also was greater among men who had been seeing their healthcare provider for a longer period (P = .01) and among men with lower perceptions of interdependence (P = .01). CONCLUSIONS: The current findings suggested that efforts to enhance trust among men who are diagnosed with prostate cancer should target African-American men, men with fewer socioeconomic resources, and men with lower perceptions of interdependence. Reasons for deterioration in trust associated with greater experience with specialty providers should be explored along with the effects of interventions that are designed to address the concerns of individuals who have greater mistrust. PMID- 19296517 TI - Liquid chromatography-tandem mass spectrometry method of loxoprofen in human plasma. AB - A rapid, sensitive and selective liquid chromatography-electrospray ionization mass spectrometric method for the determination of loxoprofen in human plasma was developed. Loxoprofen and ketoprofen (internal standard) were extracted from 20 microL of human plasma sample using ethyl acetate at acidic pH and analyzed on an Atlantis dC(18) column with the mobile phase of methanol:water (75:25, v/v). The analytes were quantified in the selected reaction monitoring mode. The standard curve was linear over the concentration range of 0.1-20 microg/mL with a lower limit of quantification of 0.1 microg/mL. The coefficient of variation and relative error for intra- and inter-assay at four quality control levels were 2.8 5.2 and 4.8-7.0%, respectively. The recoveries of loxoprofen and ketoprofen were 69.7 and 67.6%, respectively. The matrix effects for loxoprofen and ketoprofen were practically absent. This method was successfully applied to the pharmacokinetic study of loxoprofen in humans. PMID- 19296518 TI - Histidine affinity chromatography of homo-oligonucleotides. Role of multiple interactions on retention. AB - The recent application of histidine-agarose affinity supports in plasmid purification takes advantage of the biorecognition of nucleic acid bases by the histidine ligand. This consideration prompted the need for better understanding the interactions involved in affinity chromatography of plasmid DNA with the histidine-agarose support. In this work, we used synthetic homo deoxyoligonucleotides with different sizes (1-30 nucleotides long), to explore the effect of several conditions like hydrophobic character of the individual bases, presence of secondary structures, temperature, pH and salt concentration on the mechanism of retention of nucleic acids to histidine-agarose support. One of the most striking results shows that histidine interacts preferentially with guanine, and the presence of secondary structures on polyA and polyG oligonucleotides has a significant influence on retention. Otherwise, the temperature manipulation has not shown a direct influence on oligonucleotide retention, only inducing conformational changes on secondary structures. Overall, the results obtained provide valuable information for the future development and implementation of histidine and other amino acids as ligands in chromatography for the purification of plasmid DNA and other nucleic acids, by improving the knowledge of the interactions involved as well as of the parameters influencing the retention. PMID- 19296519 TI - Development of a method for the determination of vardenafil in human plasma by high performance liquid chromatography with UV detection. AB - A simple high-performance liquid chromatographic (HPLC) method with photometric detection is described for the determination of vardenafil hydrochloride, a phosphodiesterase V inhibitor, in human plasma. Chromatographic separation of the analyte and internal standard was achieved on an analytical 250 x 4.6 mm i.d. reversed-phase Kromasil KR 100 C(18) (5 microm particle size) column using a mobile phase of acetonitrile-potassium dihydrogen phosphate (30:70 v/v). The run time was less than 15 min. Column eluate was monitored at 230 nm. The linearity over the concentration range of 10-1500 ng/mL for vardenafil was obtained and the limit of quantification (LOQ) was 10 ng/mL. The method has been applied to analysis of the vardenafil concentrations for application in pharmacokinetic studies. PMID- 19296520 TI - Determination of tectoridin in rat plasma by high-performance liquid chromatography and its application to pharmacokinetic studies. AB - A simple, specific and sensitive HPLC method with UV detection was developed and validated for the determination of tectoridin in rat plasma for the first time. Chromatographic separation was performed on a Welchrom C(18) column (150 x 4.6 mm, i.d., 5 microm) at a flow rate of 1.0 mL min(-1), using a mixture of methanol 2% HAc aqueous solution (31:69, v/v) as the mobile phase with UV detection at 266 nm. The calibration curves for tectoridin were linear over the concentration range of 1.10-274.40 microg mL(-1) in rat plasma. The intra- and inter-day accuracies (RE) were within -3.23% and 4.11%. The intra- and inter-day precisions (RSD) were not more than 2.74 and 4.72%, respectively. The present method was successfully applied to the pharmacokinetic studies of tectoridin in rats after intravenous administration of three different doses. PMID- 19296521 TI - The in vivo anatomy of the deep circumflex iliac artery perforators: defining the role for the DCIA perforator flap. AB - The deep circumflex iliac artery (DCIA) provides a dependable option for use as an osteo-musculo-cutaneous flap, particularly in mandibular reconstruction. Modifications to flaps based on DCIA perforators have been sought to prevent donor site morbidity as a consequence of muscle cuff harvest. Previous studies have been inconsistent in their descriptions of perforator anatomy, and means of assessing these preoperatively have not been widely described. A clinical anatomical study was undertaken, with a cohort of 44 hemiabdominal walls in 22 consecutive patients undergoing preoperative computed tomographic angiography (CTA) before free flap surgery. The feasibility of CTA and the regional vascular anatomy were both assessed. The use of CTA was shown to demonstrate DCIA perforators with high resolution and to be able to assess vessel size and location. In 44 hemiabdominal walls, there were 44 perforators of >0.8 mm diameter. There were no suitable perforators in 40% of sides, with 32% of sides having one perforator >0.8 mm diameter, 16% having two perforators, <10% had three perforators, and only one side had over four perforators. Perforators emerged from the deep fascia on an average of 5.1 cm cranial and 3.9 cm posterior to the anterior superior iliac spine (ASIS). Of the 44 perforators identified, 82% of perforators were located within a 4 cm by 4 cm area, 3 cm superior, and 2 cm posterior to the ASIS. The current study has demonstrated the utility of preoperative CTA for identifying DCIA perforators, and for selecting patients who may be suitable for a DCIA perforator flap given the variable perforator anatomy. PMID- 19296522 TI - Denervation and reinnervation of adult skeletal muscle modulate mRNA expression of neuregulin-1 and ErbB receptors. AB - Skeletal muscle atrophy represents one of the main causes of poor outcome of microsurgical nerve reconstruction. Recent studies have pointed to the importance of the neuregulin/ErbB signaling pathway in the development and regeneration of the neuromuscular system. Here, we show by immunohistochemistry, RT-PCR, and Western blotting analyses, in an in vivo model of adult skeletal muscle denervation/reinnervation, that expression of Neuregulin1 (NRG1) and ErbB receptors is regulated by the innervation condition. We found out that a significant upregulation of the alpha-, but not beta-, isoform of NRG1, as well as of ErbB2, ErbB3, and ErbB4-cyt1 isoform occurs as a consequence of denervation of flexor digitorum muscles of the rat forelimb by median nerve transection. Moreover, after tubulization median nerve repair, and consequent muscle reinnervation, all messengers of the NRG1/ErbB system are promptly downregulated. Therefore, our results suggest the existence of a alpha-NRG1-mediated autocrine and/or paracrine trophic loop in skeletal muscles that is activated after denervation and promptly deactivated after nerve reconstruction. This myotrophic loop is a promising therapeutic target for the prevention of muscle atrophy. Yet, the recent demonstration of a similar alpha-NRG1-mediated gliotrophic loop in denervated Schwann cells provides a possible explanation for the effectiveness of muscle conduits for tubulization nerve repair. PMID- 19296523 TI - Aesthetic restoration of Poland's syndrome in a male patient using free anterolateral thigh perforator flap as autologous filler. AB - We report a case of a male patient with Poland's anomaly who was reconstructed with a free anterolateral thigh perforator flap. The flap was used successfully as an autologous filler to recreate the anterior axillary line and correct the chest contour deformity. The use of the free anterolateral thigh perforator flap is an excellent choice as an autologous filler to correct mild and moderate deformity in male Poland's syndrome, carrying low morbidity and leaving both minimal scarring and functional sequelae. PMID- 19296524 TI - Chromosomal aberrations in UVB-induced tumors of immunosuppressed mice. AB - In immunocompromised individuals, such as organ transplant recipients, the risk of cutaneous squamous cell carcinoma (SCC) is increased 60-250 fold, and there is an increased likelihood to develop aggressive, metastatic SCC. An understanding of the genes involved in SCC tumorigenesis is critical to prevent SCC-associated morbidity and mortality. Mouse models show that different immunosuppressive drugs lead to SCCs varying in size, number, and malignant potential. In this study, we used mouse models that mimic adult transplant recipients to study the effect of immunosuppressive drugs and UV light on SCC development. UV-induced tumors from six treatment groups, control, tacrolimus (Tac), rapamycin (Rap), cyclosporin (CsA), mycophenolate mofetil (MMF), and Rap plus CsA, were evaluated by array comparative genomic hybridization. Mouse SCCs appear to show similar genomic aberrations as those reported in human SCCs and offer the ability to identify genomic changes associated with specific and combinatorial effects of drugs. Fewer aberrations were seen in tumors of mice treated with MMF or Rap. Tumors from Tac-treated animals showed the highest number of changes. Calcineurin inhibitors (Tac and CsA) did not cluster together by their genomic aberrations, indicating their contribution to UV mediated carcinogenesis may be through different pathways. The combination treatment (Rap plus CsA) did not cluster with either treatment individually, suggesting it may influence SCC tumorigenesis via a different mechanism. Future studies will identify specific genes mapping to regions of aberration that are different between treatment groups to identify target pathways that may be affected by these drugs. PMID- 19296525 TI - Differential regulation of human thymosin beta 15 isoforms by transforming growth factor beta 1. AB - We recently identified an additional isoform of human thymosin beta 15 (also known as NB-thymosin beta, gene name TMSB15A) transcribed from an independent gene, and designated TMSB15B. The purpose of this study was to investigate whether these isoforms were differentially expressed and functional. Our data show that the TMSB15A and TMSB15B isoforms have distinct expression patterns in different tumor cell lines and tissues. TMSB15A was expressed at higher levels in HCT116, DU145, LNCaP, and LNCaP-LN3 cancer cells. In MCF-7, SKOV-3, HT1080, and PC-3MLN4 cells, TMSB15A and TMSB15B showed approximately equivalent levels of expression, while TMSB15B was the predominant isoform expressed in PC-3, MDA-MB 231, NCI-H322, and Caco-2 cancer cells. In normal human prostate and prostate cancer tissues, TMSB15A was the predominant isoform expressed. In contrast, normal colon and colon cancer tissue expressed predominantly TMSB15B. The two gene isoforms are also subject to different transcriptional regulation. Treatment of MCF-7 breast cancer cells with transforming growth factor beta 1 repressed TMSB15A expression but had no effect on TMSB15B. siRNA specific to the TMSB15B isoform suppressed cell migration of prostate cancer cells to epidermal growth factor, suggesting a functional role for this second isoform. In summary, our data reveal different expression patterns and regulation of a new thymosin beta 15 gene paralog. This may have important consequences in both tumor and neuronal cell motility. PMID- 19296526 TI - Spontaneous apoptosis of tumor-specific tetramer+ CD8+ T lymphocytes in the peripheral circulation of patients with head and neck cancer. AB - BACKGROUND: In cancer, tumor escape from the host immune system includes apoptosis of circulating CD3(+)CD8(+) effector T lymphocytes. Here, we compare sensitivity to apoptosis of virus- with tumor-specific circulating CD8(+) T cells in patients with head and neck cancer. METHODS: Wild-type p53 peptide-specific (p53(264-272) and p53(149-157)) and viral peptide-specific (EBV BMLF(259-267) and CMVpp65(495-503)) tetramers were used to measure the frequency of reactive T cells by flow cytometry. Annexin V (ANX) binding to circulating 7-amino actinomycin D-negative but tetramer(+)CD8(+) T cells in PBMC obtained from 21 patients with head and neck cancer and 11 normal controls (NC) was evaluated. RESULTS: In patients with head and neck cancer, a higher percentage of tetramer(+)CD8(+) than tetramer(-)CD8(+) T cells bound ANX (p < .023-.005). Although most tumor-epitope(+)CD8(+) T cells bound ANX, lower percentages of virus-specific CD8(+) T cells were ANX(+) in the same patients. CONCLUSIONS: Preferential demise of circulating tumor-specific CD8(+) T cells and their paucity in head and neck cancer contribute to tumor escape. PMID- 19296527 TI - Arteriovenous malformation of the scalp with cerebral steal. AB - BACKGROUND: Arteriovenous malformation of the scalp resulting from blunt trauma to the head is an unusual occurrence and is rarely associated with neurological sequelae. This case details an arteriovenous malformation with a cerebral steal phenomenon. METHODS: A 21-year-old man presented with an enlarging, pulsatile mass in the right facial/temporal area without known cause. CT and angiography were consistent with a large arteriovenous malformation involving the external carotid system, causing a secondary cerebral steal that resulted in neurological symptoms including seizure. Surgical resection of the mass was undertaken. RESULTS: The patient had an uncomplicated preoperative course with complete resolution of all neurologic symptoms. CONCLUSION: Cerebral steal is an uncommon consequence of traumatic extracranial arteriovenous malformation. The current literature is reviewed, and the diagnostic approach as well as therapeutic options are discussed. PMID- 19296528 TI - Prospective study of percutaneous endoscopic gastrostomy tubes versus nasogastric tubes for enteral feeding in patients with head and neck cancer undergoing (chemo)radiation. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) tubes have largely replaced nasogastric tubes (NGTs) for nutritional support of patients with head and neck cancer undergoing curative (chemo) radiotherapy without any good scientific basis. METHODS: A prospective study was conducted to compare PEG tubes and NGTs in terms of nutritional outcomes, complications, patient satisfaction, and cost. RESULTS: There were 32 PEG and 73 NGT patients. PEG patients sustained significantly less weight loss at 6 weeks post-treatment (median 0.8 kg gain vs 3.7 kg loss, p < .001), but had a high insertion site infection rate (41%), longer median duration of use (146 vs 57 days, p < .001), and more grade 3 dysphagia in disease-free survivors at 6 months (25% vs 8%, p = .07). Patient self-assessed general physical condition and overall quality of life scores were similar in both groups. Overall costs were significantly higher for PEG patients. CONCLUSION: PEG tube use should be selective, not routine, in this patient population. PMID- 19296529 TI - Free vascularized fibula grafts for salvage of failed oncologic long bone reconstruction and pathologic fractures. AB - Vascularized bone grafts, particularly the free fibula transfer, have incited revolutionary changes in the field of skeletal reconstruction. In no place has this been more evident than in oncologic reconstruction. The free vascularized fibula graft has been used to good effect for primary long-bone reconstruction, long-bone allograft complication salvage, and pathologic fracture salvage of the long bones. Although many of these procedures often entail significant complications, limb salvage has been made possible in a majority of patients using transfers of free vascularized fibula grafts. The purpose of this review is to critically evaluate the technique of onlay free vascularized fibula grafts for salvage of allograft complications and pathologic fractures of the long bones. This will be accomplished by reviewing the problem of allograft complications and pathologic fractures, the current treatment modalities available, the outcomes of these treatments, and future directions of treatment for this particular problem. PMID- 19296530 TI - Free latissimus dorsi flap with long venous grafts for closure of a soft tissue defect of the spine in a patient with Noonan's syndrome: a case report. AB - A 23-year-old male patient was operated for a soft tissue defect of the spine using a free musculocutaneous latissimus dorsi flap with two 21-cm long venous grafts connecting the flap to its vascular supply at the thoracodorsal vessels. This young patient suffered from Noonan's Syndrome that is a complex hereditary malformation syndrome with clinical features often leading to spinal deformities and gibbus development. The defect originated from a former internal vertebral fixation that was necessary for erective correction of such a spinal deformity. After perforation of the introduced material and debridement the defect was 4 x 8 cm in size, which was successfully closed by the free latissimus dorsi transfer. However, two venous grafts were necessary because the extension of the pedicled muscle was not sufficient to reach the defect. This is the first case in literature where a free flap was used with two venous grafts to close a soft tissue defect of the spine in the special case of a vertebral deformity based on a Noonan's Syndrome. PMID- 19296531 TI - Prediction of drug bioavailability in humans using immobilized artificial membrane phosphatidylcholine column chromatography and in vitro hepatic metabolic clearance. AB - This study reports a rapid screening method for the prediction of oral drug bioavailability in humans based on combined immobilized artificial membrane (IAM) chromatographic capacity factor (k(IAM)) and in vitro stability in hepatic microsomes. The fraction of drug absorbed (F(a)) in humans was predicted for a set of 15 structurally diverse commercial drugs based on k(IAM) values using a mobile phase consisting of acetonitrile: Dulbecco's phosphate-buffered saline. The hepatic intrinsic clearance (CL'(int)) was calculated from in vitro disappearance half-life, and the oral bioavailability was predicted using in vitro hepatic clearance (CL(h)) and F(a). Significant correlations were observed for the relationships between predicted hepatic extraction ratios (ER(h)) and actual presystemic metabolism (r = 0.854) and between predicted and observed oral bioavailabilities (r = 0.805; p < 0.01). The IAM capacity factor together with the hepatic microsomal disappearance half-life may be useful in identifying compounds with high oral absorption potential in early drug discovery processes. PMID- 19296532 TI - The "limited rectus sheath incisions" technique for DIEP flaps using preoperative CT angiography. AB - The abdominal wall donor site has become popularized for autologous breast reconstruction, with low donor site morbidity one of its key attributes. Further improvements in donor site outcomes have been facilitated through muscle sparing techniques, such as the use of the deep inferior epigastric artery (DIEA) perforator (DIEP) flap, and the use of modifications to closing the rectus sheath following muscular dissection, such as the use of endoscopic techniques and the use of mesh. We describe a new and unique method for potentially minimizing donor site damage during DIEP flap harvest: the "limited rectus sheath incisions" technique. This approach is only possible in select cases, the choice of which can be aided with the use of preoperative computed tomography angiography (CTA). In select patients, there may be an extended segment of DIEA that does not give any musculocutaneous perforators, and thus does not require open exposure. In such cases, limited incision to the rectus sheath may be made: one to access the periumbilical perforators and a separate incision to access the pedicle near its origin on the external iliac artery. This incision may then be made in a muscle splitting fashion, in the line of external oblique fibers. This approach may reduce damage to the anterior rectus sheath, and may contribute to improving donor site morbidity. (c) 2009 Wiley-Liss, Inc. Microsurgery, 2009. PMID- 19296533 TI - Reversed-phase liquid chromatography method for the determination of total plasma thiols after derivatization with 1-benzyl-2-chloropyridinium bromide. AB - A high-performance liquid chromatography method was developed for simultaneous detection and quantitation of total cysteine, glutathione, homocysteine and cysteinylglycine in human plasma. The two key steps in the analysis are reduction of disulfides and treatment with 1-benzyl-2-chloropyridinium bromide, which rapidly and quantitatively reacts with thiol groups to form stable S-pyridinium derivatives with intense UV absorption. The derivatives are well separated on a Zorbax SB C(18) column using reversed-phase high-performance liquid chromatography and monitored at 315 nm. The calibration graphs were linear over concentration ranges covering most experimental and clinical cases with a regression coefficients better than 0.999. The detection and quantitation limits for all analytes were 0.2 and 0.5 micromol/L, respectively. The recoveries were 99.25-101.68%. The intra- and interassay imprecisions were 0.88-4.24 and 1.68 5.14%, respectively. The method was applied for plasma samples donated by apparently healthy volunteers. PMID- 19296534 TI - Widespread changes in dendritic and axonal morphology in Mecp2-mutant mouse models of Rett syndrome: evidence for disruption of neuronal networks. AB - Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the X linked gene MECP2. Girls with RTT show dramatic changes in brain function, but relatively few studies have explored the structure of neural circuits. Examining two mouse models of RTT (Mecp2B and Mecp2J), we previously documented changes in brain anatomy. Herein, we use confocal microscopy to study the effects of MeCP2 deficiency on the morphology of dendrites and axons in the fascia dentata (FD), CA1 area of hippocampus, and motor cortex following Lucifer yellow microinjection or carbocyanine dye tracing. At 3 weeks of age, most (33 of 41) morphological parameters were significantly altered in Mecp2B mice; fewer (23 of 39) were abnormal in Mecp2J mice. There were striking changes in the density and size of the dendritic spines and density and orientation of axons. In Mecp2B mice, dendritic spine density was decreased in the FD (approximately 11%), CA1 (14 22%), and motor cortex (approximately 16%). A decreased spine head size (approximately 9%) and an increased spine neck length (approximately 12%) were found in Mecp2B FD. In addition, axons in the motor cortex were disorganized. In Mecp2J mice, spine density was significantly decreased in CA1 (14-26%). In both models, dendritic swelling and elongated spine necks were seen in all areas studied. Marked variation in the type and extent of changes was noted in dendrites of adjacent neurons. Electron microscopy confirmed abnormalities in dendrites and axons and showed abnormal mitochondria. Our findings document widespread abnormalities of dendrites and axons that recapitulate those seen in RTT. PMID- 19296535 TI - Increased levels of copper efflux transporter ATP7B are associated with poor outcome in colorectal cancer patients receiving oxaliplatin-based chemotherapy. AB - Recently, the copper efflux transporters ATP7B and ATP7A have been implicated in the transport of and resistance to platinum drugs in breast and ovarian cancers. Because of the extensive use of oxaliplatin in colorectal cancer (CRC), we examined the expression of both transporters in tumors from CRC patients treated with oxaliplatin/5FU and sought to determine whether their expression can predict clinical outcome in these patients. ATP7B and ATP7A levels were determined by quantitative real-time PCR in 50 primary tumors of previously untreated patients with advanced colorectal adenocarcinoma who were subsequently treated with oxaliplatin/5FU. Additionally, ATP7B protein expression was assessed by immunohistochemical staining using a tissue microarray. Patients with the lowest mRNA expression levels of ATP7B had a significantly longer time to progression (TTP) (p = 0.0009) than patients with the highest levels (12.14 months vs. 6.43 months) who also had an increased risk of progression (HR = 3.56; 95% CI, 1.6 7.9; p = 0.002). Furthermore, patients with low levels of both protein and mRNA of ATP7B derived the maximum benefit from oxaliplatin/5FU with the longest TTP as compared with patients with high levels of ATP7B protein and mRNA (14.64 months vs. 4.63 months, respectively, p = 0.01) and showed a nonsignificant trend toward a lower response rate (37.5% and 75%, respectively). In conclusion, ATP7B mRNA and protein expression in colorectal tumors is associated with clinical outcome to oxaliplatin/5FU. Prospective studies are required to evaluate the role of this marker in tailoring chemotherapy. PMID- 19296536 TI - Evaluation of nonviral risk factors for nasopharyngeal carcinoma in a high-risk population of Southern China. AB - To understand the role of environmental and genetic influences on nasopharyngeal carcinoma (NPC) in populations at high risk of NPC, we have performed a case control study in Guangxi Province of Southern China in 2004-2005. NPC cases (n = 1,049) were compared with 785 NPC-free matched controls who were seropositive for IgA antibodies (IgA) to Epstein-Barr virus (EBV) capsid antigen (VCA)-a predictive marker for NPC in Chinese populations. A questionnaire was used to capture exposure and NPC family history data. Risk factors associated with NPC in a multivariant analysis model were the following: (i) a first, second or third degree relative with NPC [attributable risk (AR)= 6%, odds ratio (OR) = 3.1, 95% confidence interval (CI) = 2.0-4.9, p < 0.001]; (ii) consumption of salted fish 3 or more than 3 times per month (AR = 3%, OR = 1.9, 95% CI = 1.1-3.5, p = 0.035); (iii) exposure to domestic wood cooking fires for more than 10 years (AR = 69%, OR = 5.8, 95% CI = 2.5-13.6, p < 0.001); and (iv) exposure to occupational solvents for 10 or less years (AR = 4%, OR = 2.6, 95% CI = 1.4-4.8, p = 0.002). Consumption of preserved meats or a history of tobacco smoking were not associated with NPC (p > 0.05). We also assessed the contribution of EBV/IgA/VCA antibody serostatus to NPC risk-32.2% of NPC can be explained by IgA+ status. However, family history and environmental risk factors cumulatively explained only 2.7% of NPC development in NPC high risk population. These findings should have important public health implications for NPC risk reduction in endemic regions. PMID- 19296537 TI - Cancer/testis antigen CT45: analysis of mRNA and protein expression in human cancer. AB - CT45 is a cancer/testis gene that we previously identified by massively parallel signature sequencing. Encoded by a multigene family on chromosome X, CT45 showed restricted mRNA expression to normal testis and various cancers. In this study, monoclonal antibodies were generated against recombinant CT45 protein, and CT45 protein expression in normal and tumor tissues was evaluated by immunohistochemical analysis. In adult normal tissue, CT45 expression was restricted to testicular germ cells, detected as a nuclear protein mainly at the stage of primary spermatocytes. In tumors, CT45 protein expression correlated with the mRNA levels detected by quantitative RT-PCR, and most lung cancer and ovarian cancers with CT45 mRNA at levels >1% of testicular expression were CT45 protein-positive. In positive cases, CT45 showed expression patterns that ranged from diffuse strong staining to heterogeneous and patchy expression. In lung cancer, CT45 expression was least frequent in adenocarcinoma, more frequent in squamous cell carcinoma and neuroendocrine tumors. Using tissue microarrays, 376 lung cancer, 219 ovarian cancer and 155 breast cancer were evaluated for CT45 protein expression. The expression frequency was highest in ovarian cancer (37%), followed by lung cancer (13%) and lowest in breast cancer (<5%). Given the focal nature of CT45 expression in many cases, these numbers represented the minimal frequency of expression in these tumor types. In summary, the expression frequency and characteristics of CT45 expression are similar to other CT cancer vaccine targets currently in clinical trials, e.g., NY-ESO-1 and MAGE-A, suggesting CT45 as a potentially useful cancer target. PMID- 19296538 TI - FGFR4 Gly388Arg polymorphism may affect the clinical stage of patients with lung cancer by modulating the transcriptional profile of normal lung. AB - The association of the fibroblast growth factor receptor 4 (FGFR4) Gly388Arg polymorphism with clinical stage and overall survival in a series of 541 Italian lung adenocarcinoma (ADCA) patients indicated a significantly decreased survival in patients carrying the rare Arg388 allele as compared to that in Gly/Gly homozygous patients [hazard ratio (HR) = 1.5; 95% confidence interval (CI) 1.1 1.9], with the decrease related to the association of the same polymorphism with clinical stage (HR = 1.8, 95% CI 1.3-2.6). By contrast, no significant association was detected in small series of either Norwegian lung ADCA patients or Italian lung squamous cell carcinoma (SQCC) patients. Single nucleotide polymorphisms of known FGFR4 ligands expressed in lung (FGF9, FGF18 and FGF19) were not associated with clinical stage or survival and showed no interaction with FGFR4. Analysis of gene expression profile in normal lungs according to FGFR4 genotype indicated a specific transcript pattern associated with the allele carrier status, suggesting a functional role for the FGFR4 polymorphism already detectable in normal lung. These findings confirm the significant association of the FGFR4 Gly388Arg polymorphism with clinical stage and overall survival in an Italian lung ADCA population and demonstrate a FGFR4 genotype-dependent transcriptional profile present in normal lung tissue. PMID- 19296539 TI - Effects of interferon alpha treatment on recurrence and survival after complete resection or ablation of hepatocellular carcinoma: a meta-analysis of randomized controlled trials. AB - Available literature on the benefit of interferon alpha (IFN-alpha) as adjuvant postsurgical or ablative treatment of hepatocellular carcinoma reports discordant results. By meta-analysis of the available data, we evaluated the effects of IFN alpha on recurrence and survival after complete resection or ablation of hepatocellular carcinoma. All randomized controlled trials comparing IFN-alpha with placebo or no treatment after tumor resection or ablation were selected. Finally, 6 studies published in 2001 or later with a total of 600 patients were included in this meta-analysis. Data on postsurgical or ablative early recurrence and 1 year survival of hepatocellular carcinoma in IFN-alpha treated and untreated patients were extracted from each study. Proportions were combined, and the odds ratio (OR) with its 95% confidence interval (CI) was used as the effect size estimate. Analysis results show that IFN-alpha significantly decreased postsurgical or ablative overall early recurrence (OR = 0.62; 95% CI = 0.42-0.93; p = 0.02) and improved overall 1 year survival (OR = 3.14; 95% CI = 1.79-5.52; p < 0.0001). Subgroup analyses show that IFN-alpha decreased postsurgical early recurrence (OR = 0.58; 95% CI = 0.37-0.91; p = 0.02) and improved 1 year survival (OR = 3.19; 95% CI = 1.80-5.67; p < 0.0001) evidently. Subgroup analyses also show that IFN-alpha reduced early recurrence after resection without pre resection ablation therapy (OR = 0.58; 95% CI = 0.37-0.91; p = 0.02) and improved 1 year survival (OR = 3.83; 95% CI = 2.01-7.27; p < 0.0001). These results suggest that IFN-alpha treatment could significantly decrease early recurrence and improve 1 year survival of patients with hepatocellular carcinoma after complete resection or ablation. The use of IFN-alpha as adjuvant postsurgical or ablative treatment seems promising but requires further study. (c) 2009 UICC. PMID- 19296540 TI - Side population of pancreatic cancer cells predominates in TGF-beta-mediated epithelial to mesenchymal transition and invasion. AB - We report here side population (SP) cells, a cancer stem cell enriched fraction from pancreatic cancer cell line, have enormous superior potential of the epithelial to mesenchymal transition (EMT), invasion, and metastasis. In an isolated SP cell culture, the cells rapidly expressed and up-regulated E cadherin, an epithelial phenotypic marker, and the cells formed tightly contacted cell cluster, which is a representative epithelial phenotypic appearance. When the SP cells were incubated in the presence of TGF-beta, SP cells changed their shape into mesenchymal-like appearance including spindle shaped assembly. This alteration was associated with significant reduction of E-cadherin expression level. TGF-beta induced EMT-associated gene alteration such as reduction of E cadherin mRNA and induction of Snail mRNA and matrixmetalloproteinase (MMP)-2 mRNA. Finally, SP cells exerted notable matrigel invasion activity in response to TGF-beta treatment, whereas MP cells did not respond to TGF-beta-mediated invasion. In conclusion, these results suggest that SP cells from pancreatic cancer cell line possess superior potentials of phenotypic switch, i.e., EMT/MET, micro-invasion, and in vivo metastasis, as compared to MP cells. Because micro invasion and metastasis are key mechanisms of cancer malignant potential, SP cells would be the attractive target for preventing cancer progression. PMID- 19296541 TI - Chondrogenesis of human mesenchymal stem cells encapsulated in a hydrogel construct: neocartilage formation in animal models as both mice and rabbits. AB - In this study, in vivo studies, both nude mouse and rabbit cartilage defect, were tested for chondrogenesis using stem cells (SCs) using growth factor. Specifically, human mesenchymal stem cells (hMSCs) were embedded in a hydrogel scaffold, which was coencapsulated with transforming growth factor-beta3 (TGF beta3). The specific extracellular matrices (ECMs) released from hMSCs transplanted into the animal were assessed via glycosaminoglycan (GAG)/DNA content, RT-PCR, real time-QPCR, immunohistochemical (IHC), and Safranin-O staining and were observed up to 7 weeks after injection. By detection of ECMs the GAG content per cell remained constant for all formulations, indicating that the dramatic increase in cell number for samples with TGF-beta3 was accompanied by the maintenance of the cell phenotypes. The histological and IHC staining of the newly repaired tissues observed after treatment with TGF-beta3 mixed with hMSCs evidenced hyaline cartilage-like characteristics. Moreover, the results observed with the animal model (rabbit) treated with hMSCs embedded in the growth factor-containing hydrogel indicate that the implantation of mixed cells with TGF beta3 may constitute a clinically efficient method for the regeneration of hyaline articular cartilage. PMID- 19296542 TI - Plasma-sprayed hydroxyapatite coating on carbon/carbon composite scaffolds for bone tissue engineering and related tests in vivo. AB - The bioactive hydroxyapatite (HA) coatings were successfully prepared on carbon/carbon composites (C/C) by means of sand-blasting pretreatment and plasma spraying technology. X-ray diffraction was employed to analyze the phase constitute of the coatings. Meanwhile, the bond strength between the HA coatings and C/C substrates was determined via shear test. Experimental results show that the coatings constitute HA, CaO, and other amorphous phosphates. The post heat treatment could effectively increase crystallization and purity of the coatings. Through observation and analysis by electron microprobe and scanning electron microscopy, it is concluded that the bond strength of the plasma-sprayed HA coatings on C/C is mainly determined by the interface structure and can be further improved by the post heat treatment. Meanwhile, the implantation in vivo was carried out in hybrid goats. The histological observation revealed that the osteoplaque gradually grew on the surface of the HA coatings and the pure C/C surface was covered by the fibrous tissues. No inflammation symptoms were found in the bone tissue around the implants. PMID- 19296543 TI - Functionally graded beta-TCP/PCL nanocomposite scaffolds: in vitro evaluation with human fetal osteoblast cells for bone tissue engineering. AB - The engineering of biomimetic tissue relies on the ability to develop biodegradable scaffolds with functionally graded physical and chemical properties. In this study, a twin-screw-extrusion/spiral winding (TSESW) process was developed to enable the radial grading of porous scaffolds (discrete and continuous gradations) that were composed of polycaprolactone (PCL), beta tricalciumphosphate (beta-TCP) nanoparticles, and salt porogens. Scaffolds with interconnected porosity, exhibiting myriad radial porosity, pore-size distributions, and beta-TCP nanoparticle concentration could be obtained. The results of the characterization of their compressive properties and in vitro cell proliferation studies using human fetal osteoblast cells suggest the promising nature of such scaffolds. The significant degree of freedom offered by the TSESW process should be an additional enabler in the quest toward the mimicry of the complex elegance of the native tissues. PMID- 19296544 TI - Novel template-casting technique for fabricating beta-tricalcium phosphate scaffolds with high interconnectivity and mechanical strength and in vitro cell responses. AB - A novel template-casting method was developed to produce completely interconnected, macroporous biodegradable beta-tricalcium phosphate (beta-TCP) scaffolds, whose architecture and chemistry can be fully manipulated by varying the templates and casting materials. The processing route includes preparation of beta-TCP slurry; casting and shaping into preformed templates comprised of paraffin beads; solidifying, drying; and sintering. Structural, chemical, and mechanical properties of the prepared macroporous scaffolds were characterized using micro computed tomography, scanning electron microscopy, x-ray diffractometry, Fourier transform infrared spectroscopy, and mechanical testing. Human embryonic palatal mesenchymal cells were used to evaluate cell proliferation within the scaffolds in vitro. The scaffolds consisted of interconnected macropores and solid struts, leading to a reticular network. Two groups of scaffolds with larger pores, approximately 600-800 microm and smaller pores approximately 350-500 microm, were demonstrated. The interconnected windows between neighboring macropores were 440 +/- 57 microm in diameter for the larger pored scaffolds, and 330 +/- 50 microm for the smaller-pored scaffolds. The scaffolds were highly crystallized and composed dominantly of beta-tricalcium phosphate (beta-TCP) accompanied by minor phase of hydroxyapatite (HA). The hydroxyl group was clearly detected by FTIR on the scaffolds. High mechanical strength (9.3 MPa) was demonstrated by the completely interconnected scaffolds with approximately 79% porosity. The human embryonic palatal mesenchymal (HEPM) cells proliferated well on the smaller-pored and larger-pored scaffolds, exhibiting a significantly higher level of proliferation in the first 11 days of culture on the smaller pored scaffolds. High levels of differentiation were also evidenced in both pore sizes of scaffolds. PMID- 19296545 TI - Fibronectin-DNA-apatite composite layer for highly efficient and area-specific gene transfer. AB - In 2004, Shen et al. developed a safe and efficient gene transfer system using a DNA-apatite composite layer. We have recently succeeded in improving further the gene transfer efficiency by immobilizing a cell adhesion molecule laminin, in a DNA-apatite composite layer. In this study, we showed that not only laminin but fibronectin immobilized in a DNA-apatite composite layer enhances cell adhesion and cell spreading on the layer, thereby markedly improving the gene transfer efficiency. Therefore, the immobilization of a cell adhesion molecule in a DNA apatite composite layer is crucial for improving the gene transfer efficiency. By using fibronectin instead of laminin and optimizing the condition to prepare the fibronectin-DNA-apatite composite layer, the amount (weight) of cell adhesion molecule required was reduced to approximately one-fourth while retaining the relatively high gene transfer efficiency. It was also shown that the resulting fibronectin-DNA-apatite composite layer prepared under the optimized condition mediated the area-specific gene transfer on its surface, that is, DNA was preferentially transferred to the cells adhering to the surface of the fibronectin-DNA-apatite composite layer. The present gene transfer system with potential for area-specific transfection and advantages of safety and relatively high efficiency would be useful in tissue engineering applications, gene therapy, and production of transfection microarrays. PMID- 19296546 TI - Measuring the strength of dermal fibroblast attachment to functionalized titanium alloys in vitro. AB - The success of intraosseous transcutaneous amputation prostheses (ITAP) relies on soft tissue attachment to prevent infection which leads to implant failure. Fibronectin (Fn) has been shown to enhance dermal fibroblast attachment in vitro, however measurement of cell attachment strength has been indirect; using cell area and immunolocalization of focal adhesion components. In this study, we have developed a flow apparatus to assess the biophysical strength of cell attachment to biomaterials used in ITAP. We have demonstrated that dermal fibroblast attachment strength increases significantly up to 96 h and that data from direct and indirect methods of assessing cell attachment strength have a significant positive correlation. Additionally, we have used direct and indirect assessment methods to demonstrate that dermal fibroblast attachment strength is significantly greater on fibronectin-coated titanium alloy compared with uncoated controls at 1, 4, and 24 hours. PMID- 19296547 TI - The human urinary proteome reveals high similarity between kidney aging and chronic kidney disease. AB - Aging induces morphological changes of the kidney and reduces renal function. We analyzed the low molecular weight urinary proteome of 324 healthy individuals from 2-73 years of age to gain insight on human renal aging. We observed age related modification of secretion of 325 out of over 5000 urinary peptides. The majority of these changes were associated with renal development before and during puberty, while 49 peptides were related to aging in adults. We therefore focussed the remainder of the study on these 49 peptides. The majority of these 49 peptides were also markers of chronic kidney disease, suggesting high similarity between aging and chronic kidney disease. Blinded evaluation of samples from healthy volunteers and diabetic nephropathy patients confirmed both the correlation of biomarkers with aging and with renal disease. Identification of a number of these aging-related peptides led us to hypothesize that reduced proteolytic activity is involved in human renal aging. Finally, among the 324 supposedly healthy individuals, some had urinary aging-related peptide excretion patterns typical of an individual significantly older than their actual age. In conclusion, these aging-related biomarkers may allow noninvasive detection of renal lesions in healthy persons and show high resemblance between human aging and chronic kidney disease. This similarity has to be taken into account when searching for biomarkers of renal disease. PMID- 19296548 TI - Identification of target antigens of self-reactive IgG in intravenous immunoglobulin preparations. AB - Intravenous immunoglobulin (IVIg) contains a wide range of self-reactive immunoglobulins (Ig) G. Acidic pH is known to increase the reactivity of purified IgG with self-antigens. We describe here the target antigens of IgG autoantibodies in IVIg and analyze the influence of acidic pH on IgG reactivities. We used 2-DE and immunoblotting with protein extracts of human umbilical vein endothelial cells (HUVEC) and HEp-2 cells. Two IVIg preparations obtained by ethanol fractionation [one with an acidic pH step (acidic-IVIg) and one with beta-propiolactone (propiolactone-IVIg)] and a pool of sera from 12 healthy individuals were tested. Serum IgG of 3 healthy individuals and IgG purified from the same sera with elution at pH 2.8 were also tested individually. Finally, propiolactone-IVIg was acidified at pH 2.8. IgG obtained with a step at low pH recognized many more target spots than IgG obtained without acidic pH. Our findings demonstrate that an acidic pH step artificially enlarges the repertoire of self-reactive IgG. Thus, protein spots recognized by IgG in propiolactone-IVIg represent the core set of self-antigens targeted by IVIg. Overall, 96 proteins were identified by MS. Fourteen were recognized in both extracts including glycolysis proteins such as alpha-enolase, RNA processing and cytoskeletal proteins such as lamin-A/C. PMID- 19296549 TI - Plasma proteome analysis for anti-obesity and anti-diabetic potentials of chitosan oligosaccharides in ob/ob mice. AB - Altered levels of adipokines, derived as a result of distorted adipocytes, are the major factors responsible for changing biochemical parameters in obesity that leads to the development of metabolic disorders such as insulin resistance and atherosclerosis. In our previous reports, chitosan oligosaccharides (CO) were proved to inhibit the differentiation of 3T3-L1 adipocytes. In the present study, an attempt was made to investigate the anti-obesity and anti-diabetic effect of CO on ob/ob mice, by means of differential proteomic analysis of plasma. This was followed by immunoblotting, and gene expression in adipose tissue to clarify the molecular mechanism. CO treatment showed reduced diet intake (13%), body weight gain (12%), lipid (29%) and glucose levels (35%). 2-DE results showed differential levels of five proteins namely RBP4, apoE, and apoA-IV by >2-fold down-regulation and by >2-fold of apoA-I and glutathione peroxidase (GPx) up regulation after CO treatment. Immunoblotting studies of adiponectin and resistin showed amelioration in their levels in plasma. Furthermore, the results of gene expressions for adipose tissue specific TNF-alpha, and IL-6 secretary molecules were also down-regulated by CO treatment. Gene expressions of PPAR gamma in adipose tissue were in good agreement with the ameliorated levels of adipokines, thereby improving the pathological state. Taken together, CO might act as a potent down-regulator of obesity-related gene expression in ob/ob mice that may normalize altered plasma proteins to overcome metabolic disorders of obesity. PMID- 19296550 TI - Asbestos fibers in para-aortic and mesenteric lymph nodes. AB - BACKGROUND: Asbestos fibers are known to accumulate in lung parenchyma and thoracic lymph nodes, but their presence and translocation into the extrapulmonary tissues need clarification. We assessed the presence of asbestos in the para-aortic (PA) and mesenteric (ME) lymph nodes. METHODS: PA and ME lymph nodes and lung tissue from 17 persons who underwent medicolegal autopsy for suspicion of asbestos-related disease and from five controls were analyzed for asbestos fibers using transmission electron microscopy. RESULTS: High concentrations of amphibole asbestos fibers were detected in several lung tissue samples and in the respective PA and ME lymph nodes. The mean concentration for the 10 persons with a lung asbestos content of >/=1 million fibers/g of dry tissue (f/g) was 0.85 (<0.05-4.36) million f/g in the PA lymph nodes and 0.55 (<0.02-2.86) million f/g in the ME lymph nodes. The respective mean values for the 12 persons with a lung asbestos concentration of <1 million f/g were 0.07 for the PA lymph nodes and 0.03 million f/g for the ME nodes. The lung asbestos burden that predicted the detection of asbestos in abdominal lymph nodes was 0.45 million f/g. CONCLUSIONS: In addition to their accumulation in lung tissue, asbestos fibers also collect in the retroperitoneal and the mesenteric lymph nodes. Even low-level occupational exposure results in the presence of crocidolite, amosite, anthophyllite, tremolite, or chrysotile in these abdominal lymph nodes. Our results support the hypothesis of lymph drainage as an important translocation mechanism for asbestos in the human body. PMID- 19296551 TI - The neural correlates of naming and fluency deficits in Alzheimer's disease: an FDG-PET study. AB - OBJECTIVE: To examine the neural processes associated with language deficits in Alzheimer's disease (AD), and in particular to elucidate the correlates of confrontation naming and word retrieval impairments. METHODS: Sixty patients with probable AD were included. Confrontation naming was assessed using the number of words spontaneously named correctly on the Boston Naming Test. We recorded the number of additional words stated following phonemic cuing. We also assessed phonemic (FAS) and semantic (supermarket items) fluency. We then correlated performance on each measure with resting cortical metabolic activity using FDG PET images. RESULTS: We found that poorer ability to spontaneously name an object was associated with hypometabolism of bilateral inferior temporal lobes. In contrast, when a phonemic cue was provided, successful naming under this condition was associated with higher metabolic activity in bilateral inferior frontal gyrus (IFG), right superior frontal gyrus (SFG), left temporal, and occipital regions. Consistent with these findings, we found that poorer semantic fluency was associated with hypometabolism in regions including both IFG and temporal regions, and poorer phonemic fluency was associated with hypometabolism in only left IFG. Across analyses, measures that required cued retrieval were associated with metabolism in the left IFG, whereas measures taxing semantic knowledge were associated with metabolic rate of left temporal cortex. CONCLUSIONS: Naming deficits in AD reflect compromise to temporal regions involved in the semantic knowledge network, and frontal regions involved in the controlled retrieval of information from that network. PMID- 19296552 TI - The Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) is associated with informant stress. AB - OBJECTIVE: To study the association between informant stress and appraisal of patients' cognitive functioning as reported by the Informant Questionnaire on Cognitive Decline in the Elderly--IQCODE. METHODS: Routinely collected data from a geriatric outpatient department (207 dyads) during the years 1995-1998 were analysed. Relative stress scale (RSS) has been categorised for possible low, intermediate and high risk of psychiatric morbidity and caregivers were combined to four groups (female and male spouses and female and male non-spouses, respectively). The relationship between IQCODE (dependent) and categorised RSS and informant groups and patient age was further studied by means of the general linear model (GLM-UNIANOVA). RESULTS: In general, spouses reported better cognitive functioning than non-spouses. There was a significant association between IQCODE and RSS (p < 0.001), and the composite variable informant group and informant gender (p < 0.001). The main effect of the interaction term RSS x informant group + informant gender was not significant. Post hoc test, however, revealed a significant effect of the interaction term RSS x female spouses (p < 0.001) on IQCODE. CONCLUSION: IQCODE is associated with informant stress. Categorisation of RSS score into groups of low, intermediate and high risk for psychiatric morbidity can be a valuable contribution to a more meaningful application of RSS in general practice. PMID- 19296553 TI - The COMT Val158Met polymorphism and cognition in depressed and nondepressed older adults. AB - OBJECTIVE: The objective of the current study was to examine the relationship between the COMT Val(158)Met polymorphism and neuropsychological performance in depressed and nondepressed older adults. METHODS: One hundred and twenty-six clinically depressed older adults and 105 nondepressed comparison participants were compared on neuropsychological performance and COMT Val(158)Met (Val/Val, Val/Met, Met/Met). RESULTS: Based on multivariate regression models, the COMT Val(158)Met polymorphism was not associated with cognitive performance among depressed or nondepressed individuals, nor did this polymorphism account for the fact that depressed individuals performed worse than nondepressed individuals on several neuropsychological tests that are typically affected by depression. There was also no difference in frequency of the COMT Val(158)Met alleles between depressed and nondepressed individuals. CONCLUSIONS: Although the current study found no association between COMT Val(158)Met polymorphism on a number of clinical neuropsychological tests that are typically found to be sensitive to depression, differential effects of the COMT Val(158)Met polymorphism on dopamine transmission in psychiatric and non-psychiatric populations may be further clarified by clinical research with neuroscience-based paradigms that segregate cognitive tasks into component processes with precise neural substrates, particularly with respect to the complex functions of the prefrontal cortex. Negative results can be important to narrowing down target processes and understanding the influence of clinical and demographic characteristics in studies of psychiatric genetics. PMID- 19296554 TI - Size effect on cell uptake in well-suspended, uniform mesoporous silica nanoparticles. PMID- 19296555 TI - Controlled degradation of DNA capsules with engineered restriction-enzyme cut sites. PMID- 19296556 TI - Mechanism for a directional, processive, and reversible DNA motor. PMID- 19296557 TI - Design of synthetic polymer nanoparticles that capture and neutralize a toxic peptide. AB - Designed polymer nanoparticles (NPs) capable of binding and neutralizing a biomacromolecular toxin are prepared. A library of copolymer NPs is synthesized from combinations of functional monomers. The binding capacity and affinity of the NPs are individually analyzed. NPs with optimized composition are capable of neutralizing the toxin even in a complex biological milieu. It is anticipated that this strategy will be a starting point for the design of synthetic alternatives to antibodies. PMID- 19296558 TI - Fabrication of monolithic bridge structures by vacuum-assisted capillary-force lithography. PMID- 19296559 TI - Porous alumina with shaped pore geometries and complex pore architectures fabricated by cyclic anodization. PMID- 19296560 TI - Synthesis of cell-laden alginate hollow fibers using microfluidic chips and microvascularized tissue-engineering applications. PMID- 19296561 TI - Doping single-layer graphene with aromatic molecules. PMID- 19296562 TI - Direct patterning of anti-human serum albumin antibodies on aldehyde-terminated silicon nitride surfaces for HSA protein detection. PMID- 19296563 TI - Preparation of monodisperse biodegradable polymer microparticles using a microfluidic flow-focusing device for controlled drug delivery. AB - Degradable microparticles have broad utility as vehicles for drug delivery and form the basis of several therapies approved by the US Food and Drug Administration. Conventional emulsion-based methods of manufacturing produce particles with a wide range of diameters (and thus kinetics of release) in each batch. This paper describes the fabrication of monodisperse, drug-loaded microparticles from biodegradable polymers using the microfluidic flow-focusing (FF) devices and the drug-delivery properties of those particles. Particles are engineered with defined sizes, ranging from 10 microm to 50 microm. These particles are nearly monodisperse (polydispersity index = 3.9%). A model amphiphilic drug (bupivacaine) is incorporated within the biodegradable matrix of the particles. Kinetic analysis shows that the release of the drug from these monodisperse particles is slower than that from conventional methods of the same average size but a broader distribution of sizes and, most importantly, exhibit a significantly lower initial burst than that observed with conventional particles. The difference in the initial kinetics of drug release is attributed to the uniform distribution of the drug inside the particles generated using the microfluidic methods. These results demonstrate the utility of microfluidic FF for the generation of homogenous systems of particles for the delivery of drugs. PMID- 19296564 TI - Synthesis of catalytically active porous platinum nanoparticles by transmetallation reaction and proposition of the mechanism. AB - A facile method for the synthesis of porous platinum nanoparticles by transmetallation reactions between sacrificial nickel nanoparticles and chloroplatinic acid (H(2)PtCl(6)) in solution, as well as at the constrained environment of the air-water interface, using a Langmuir-Blodgett instrumental setup is presented. To carry out the transmetallation at the air-water interface hydrophobized nickel nanoparticles are assembled as a monolayer on the sub phase containing platinum ions. The porous Pt nanoparticles obtained as a result of the reaction are found to act as extremely good catalysts for hydrogenation reaction. The products are well characterized by TEM, HRTEM, EDAX, and STEM. Attempts are made to postulate the plausible mechanism of this reaction to generate this kind of nanoparticle with controllable geometric shape and structure. This simple strategy has the potential to synthesize other nanomaterials of interest too. PMID- 19296565 TI - Heterogeneity in epoxy nanocomposites initiates crazing: significant improvements in fatigue resistance and toughening. PMID- 19296566 TI - Shaping colloidal rutile into thermally stable and porous mesoscopic titania balls. AB - High crystallinity and controlled porosity are advantageous for many applications such as energy conversion and power generation. Despite many efforts in the last decades, the direct synthesis of organic-inorganic composite materials with crystalline transition metal oxides is still a major challenge. In general, molecules serve as inorganic precursors and heat treatment is required to convert as-synthesized amorphous composites to stable crystalline materials. Herein, an alternative approach to the direct synthesis of crystalline polymer-metal oxide composites by using a spherical polyelectrolyte brush as the template system is presented. Pre-synthesized electrostatically stabilized rutile nanocrystals that carry a positive surface charge are used as inorganic precursors. In this approach, the strong Coulomb interactions between anionic polyelectrolyte brush chains and cationic crystalline rutile colloids, whose surfaces are not capped and therefore reactive, are the key factors for the organic-inorganic crystalline composite formation. Stepwise calcination first under argon and followed with a second calcination in air lead to the complete removal of the polymer template without collapse and porous rutile balls are obtained. The results suggest that any colloids that carry a surface charge might serve as inorganic precursors when charged templates are used. It is expected that this hierarchical route for structuring oxides at the mesoscale is generally applicable. PMID- 19296567 TI - Ionic transport through single solid-state nanopores controlled with thermally nanoactuated macromolecular gates. PMID- 19296568 TI - Temperature compensation for hybrid devices: kinesin's Km is temperature independent. PMID- 19296569 TI - On the strength of the hydrogen-carbon interaction as deduced from physisorption. AB - We deduce a new value for the potential well depth for the C-H2 interaction on the basis of experimental validations of isotherms of H2 and D2 predicted using independently characterized microstructural parameters. We use two carbons, one an activated carbon fiber whose structure has been recently characterized by us (Nguyen, T. X.; cohaut, N.; Bae, J.-S.; Bhatia, S. K. Langmuir 2008, 24, 7912) using hybrid reverse Monte Carlo simulation (HRMC) and the other the commercial Takeda 3A carbon molecular sieve whose pore size distribution is determined here from the 273 K CO2 adsorption isotherm. The conventional grand canonical Monte Carlo simulation technique incorporating a semiclassical Feynman and Hibbs (FH) potential approximation (FHGCMC) as well as path integral Monte Carlo calculations is employed to determine theoretical adsorption isotherms. It is found that curvature enhances the well depth for the LJ C-H2 interaction by a factor of 1.134 over that for a flat graphite surface, consistent with our recent study (Nguyen, T. X.; cohaut, N.; Bae, J.-S.; Bhatia, S. K. Langmuir 2008, 24, 7912). A value of the C-C well depth of 37.26 K, used for estimating the C-H2 well depth in conjunction with the Berthelot rules, with the Steele C-C well depth used for interaction with heavier gases (Ar, CO2 and CH4), leads to excellent agreement with experimental isotherms in all cases. PMID- 19296570 TI - The weak help the strong: low-molar-mass organogelators harden bitumen. AB - Low-molar-mass organogelators (LMOG) can turn liquids into thermoreversible gels because they self-assemble into a fibrous network. In contrast, using the same kind of low-molar-mass additives to harden materials, which are already solidlike on their own, has been hardly exploited. We show here that simple dicarboxylic acids are very efficient low-molar-mass organogelators (LMOG) for bitumen. Indeed, they increase the range of temperature where bitumen is a solid. Moreover, the hardness and elastic modulus of bitumen at room temperature are also improved. This concept of improving the mechanical properties of a solid with an LMOG can probably be applied to other materials. PMID- 19296572 TI - Emission enhancement and chromism in a salen-based gel system. AB - A new salicylideneaniline-based organogelator has been synthesized, and it can gelatinize organic solvents, including cyclohexane, toluene, benzene, and some mixed solvents. SEM images show that it has self-assembled into 1-D nanofibers, which further cross-link to form 3-D network. On the basis of the results of small-angle XRD and the optimized molecular length by semiempirical quantum calculations, the gelators are supposed to pack into a unimolecular lamellar structure with a period of 3.01 nm. Significantly, reversible chromism is realized with respect of the tautomerism between the NH and OH forms during the sol-gel freezing repetition. Furthermore, the gel can emit intense green light, and the fluorescent quantum yield of the gel is approximately 600 times higher than that of the solution. The aggregation-induced emission enhancement is ascribed to the formation of J aggregation and the inhibition of intramolecular rotation in the gel state. PMID- 19296573 TI - The stem cell marker CD133 (prominin-1) is phosphorylated on cytoplasmic tyrosine 828 and tyrosine-852 by Src and Fyn tyrosine kinases. AB - CD133 (prominin-1) is a transmembrane glycoprotein expressed at the surface of normal and cancer stem cells, progenitor cells, rod photoreceptor cells, and a variety of epithelial cells. Although CD133 is widely used as a marker of various somatic and putative cancer stem cells, its contribution to fundamental properties of stem cells such as self-renewal and differentiation remains unknown. CD133 contains a short C-terminal cytoplasmic domain with five tyrosine residues, including a consensus tyrosine phosphorylation site that has not yet been investigated. In this study, we show that CD133 is phosphorylated in human medulloblastoma D283 and Daoy cells, in a Src family kinase-dependent manner. The cytoplasmic domain of CD133 is tyrosine phosphorylated in Daoy cells overexpressing Src and Fyn tyrosine kinases, as well as in vitro using recombinant proteins. Deletion of the C-terminal cytoplasmic domain of CD133 considerably reduced its phosphorylation by Src. To identify the tyrosine phosphorylation sites in CD133, we used matrix-assisted laser desorption/ionization quadrupole time-of-flight (MALDI Q-TOF) and liquid chromatography tandem mass spectrometry (LC-MS/MS). Analysis of tyrosine phosphorylated CD133 by mass spectrometry and site-directed mutagenesis identified tyrosine-828 and the nonconsensus tyrosine-852 as the major tyrosine phosphorylation sites both in vitro and in intact cells. Identification of CD133 as a substrate for Src-family tyrosine kinases suggests that the cytoplasmic domain of CD133 might play an important role in the regulation of its functions. PMID- 19296574 TI - Processing scale-up of sicklepod (Senna obtusifolia L.) seed. AB - Sicklepod (Senna obtusifolia L.) is an invasive weed species especially of soybean and other field crops in the southeastern United States. The seeds contain a small amount (5-7%) of a highly colored fat as well as various phenolics, proteins, and galactomannans. The color of sicklepod seed oil is such that the presence of a small amount of the weed seed in a soybean crush lowers the quality of the soybean oil. Sicklepod is very prolific, and even volunteer stands yield >1000 lb of seed per acre, and prudence calls for tapping the potential of this weed as an alternative economic crop in the affected region. Pursuant to this, we have shown in laboratory-scale work the feasibility of separating the components of sicklepod seed. However, at kilogram and higher processing quantities, difficulties arise leading to modification of the earlier approach in order to efficiently separate components of the defatted seed meal. In a version for cleanly separating the proteins, the defatted meal was extracted with 0.5 M NaCl solution to remove globular proteins. Prolamins were extracted from the pellet left after salt extraction using 80% ethanol, and glutelins were then obtained in 0.1 N alkali from the residual solids left from ethanol treatment. In a pilot-scale version for water-soluble polysaccharides, the defatted meal was stirred with deionized water (DI) and centrifuged. The pooled centrifugates were heated to 92 degrees C (20-25 min), filtered, cooled to room temperature, and passed through a column of Amberlite XAD-4 to separate the polysaccharides from the anthraquinones. Senna obtusifolia L. is a one-stop-shop of a seed (from food components to medicinals). PMID- 19296575 TI - Does an extract of carob (Ceratonia siliqua L.) have chemopreventive potential related to oxidative stress and drug metabolism in human colon cells? AB - Phenolic ingredients of an aqueous carob extract are well characterized and consist of mainly gallic acid (GA). In order to assess possible chemopreventive mechanisms of carob, which can be used as a cacao substitute, effects on expression of genes related to stress response and drug metabolism were studied using human colon cell lines of different transformation state (LT97 and HT29). Stress-related genes, namely catalase (CAT) and superoxide dismutase (SOD2), were induced by carob extract and GA in LT97 adenoma, but not in HT29 carcinoma cells. Although corresponding protein products and enzyme activities were not elevated, pretreatment with carob extract and GA for 24 h reduced DNA damage in cells challenged with hydrogen peroxide (H(2)O(2)). In conclusion, carob extract and its major phenolic ingredient GA modulate gene expression and protect colon adenoma cells from genotoxic impact of H(2)O(2). Upregulation of stress-response genes could not be related to functional consequences. PMID- 19296576 TI - Crystallization and polymorphism of triacylglycerols contribute to the rheological properties of processed cheese. AB - The thermal, rheological, and structural behaviors of a spreadable processed cheese were studied by complementary techniques including differential scanning calorimetry (DSC), rheology, and X-ray diffraction as a function of temperature. In this product, fat is present as a dispersed phase. Thermal and rheological properties were studied at different cooling rates between 0.5 and 10 degrees C min(-1) from 60 to 3 degrees C. Crystallization properties of fat were monitored at a cooling rate of -2 degrees C min(-1) from 60 to -10 degrees C. Fat triacylglycerols (TGs) crystallized at 15 degrees C in a triple-chain length 3Lalpha (72 A) structure correlated to exothermic events and to the sudden increase in the rheological moduli G' and G''. Upon heating at 2 degrees C min( 1), the polymorphic transition of TGs evidence the melting of the 3Lalpha structure and the formation of a 2Lbeta' (36.7-41.5 A) structure. Melting of the latter follows. These transformations coincide with thermal events observed by DSC and the decrease in two steps of the rheological moduli. The influence of fat crystallization, melting, and polymorphism upon the viscoelastic properties is clearly demonstrated upon both heating and cooling. PMID- 19296577 TI - Arsenic accumulation and speciation in maize as affected by inoculation with arbuscular mycorrhizal fungus Glomus mosseae. AB - Effects of inoculation with arbuscular mycorrhizal (AM) fungus (Glomus mosseae) on arsenic (As) accumulation and speciation in maize were investigated by using As spiked soil at the application levels of 0, 25, 50, and 100 mg kg(-1). Inorganic As was the major species in plants, and mycorrhizal inoculation generally decreased concentrations of arsenite [As(III)] in maize roots and concentrations of As(III) and arsenate [As(V)] in the shoots. Dimethylarsenic acid (DMA) concentrations (detected in every plant sample) were higher in maize shoots for mycorrhizal than for nonmycorrhizal treatment, but no significant differences were observed for roots. Monomethylarsenic acid (MMA) was only detected in roots with mycorrhizal colonization. The uptake of As(V) was much lower by excised mycorrhizal than nonmycorrhizal roots, and the differences for the uptake of As(III) were negligible. Arsenate reductase (AR) activity was detected in maize roots, and it was reduced with mycorrhizal inoculation. Activities of peroxidase (POD) and superoxide dismutase (SOD) were detected in both maize shoots and roots, and they were suppressed by mycorrhizal inoculation. AM inoculation inhibited the uptake of As(V) and its reduction to As(III), reducing oxidation stress and thereby alleviating As toxicity to the host plant. PMID- 19296578 TI - Mechanism of the chemical composition changes of yuba prepared by a laboratory processing method. AB - Yuba is a filmlike soybean food made from heated soymilk that contains oil bodies (average diameter, 270 nm), particulate protein (>40 nm; average diameter, 70 nm), soluble protein (<40 nm), and carbohydrate (molecular size). Three varieties of soybean were used to make yuba. The carbohydrate in the remaining soymilk increased sharply while lipid increased a little. The particle size distributions of oil body showed the trend that smaller oil bodies were concentrated in the remaining soymilk, and the percentage of soluble protein in whole protein increased in the remaining soymilk. These results could be explained well with diffusion theory. The temperature gradient and concentration gradient originating from the heat treatment were considered to cause the net particle diffusion from the surface to the bottom soymilk. Lipids, which mainly exist as oil bodies, are easily incorporated into yuba films because a few of these less dense droplets diffuse downward, causing the lipid concentration in the soymilk to change a little. Carbohydrate at the surface quickly diffuses downward, causing the carbohydrate concentration increase in the soymilk beneath the developing yuba. Protein (particulate and soluble) in the soymilk was intermediate between lipid and carbohydrate. PMID- 19296579 TI - Differentially-expressed genes in pig Longissimus muscles with contrasting levels of fat, as identified by combined transcriptomic, reverse transcription PCR, and proteomic analyses. AB - Intramuscular fat content is important for many meat quality parameters. This work is aimed at identifying functional categories of genes associated with natural variation among individuals in intramuscular fat content to help the design of genetic schemes for high marbling potential. Taking advantage of the global nature of transcriptomic and proteomic technologies, 40 genes were identified as differently expressed between high fat and low fat pig Longissimus muscles at slaughter weight. They are involved in metabolic processes, cell communication, binding, and response to stimulus. Using real-time PCR in muscle biopsies taken earlier in the fattening period, the group with a high intramuscular fat content was also characterized by the down-expression of genes playing a negative role in adipogenesis, such as architectural transcription factor high-motility hook A1, mitogen activated protein-kinase14, and cyclin D1. These results suggest that interindividual variability in intramuscular fat content might arise essentially from differences in early adipogenesis. PMID- 19296580 TI - Alternate arrangements of two different metals at chemically-equivalent binding sites on a circle. AB - Two different metal ions, monovalent M(+) (M = Ag, Cu) and divalent Hg(2+), were alternately arranged on a circle with a diameter of 1.5 nm in such a way that six metal ions were put between two disk-shaped hexamonodentate template ligands (L), thereby forming the sandwich-shaped hexanuclear complexes [M(3)Hg(3)L(2)](9+). In spite of the fact that all six binding sites are completely equivalent, the arrangement of the two kinds of monovalent and divalent metal ions is precisely controlled to minimize electrostatic interactions between the adjacent metal ions. PMID- 19296581 TI - Capturing In+ monomers in a neutral weakly coordinating environment. AB - The application of a new bis(imino)pyridine ligand allowed the isolation and characterization of [{2,4-(t)Bu(2)C(6)H(3)N=CPh}(2)(NC(5)H(3))]In(+)(OTf)(-) as the first low-valent, main-group metal complex of this ligand scaffold. Structural analysis revealed a unique monomeric In(I) species with a surprisingly long metal-ligand bond. In conjunction with a density functional theory investigation, this complex is shown to display only nominal donor-acceptor interactions between the metal and the neutral ligand. The mixing of the occupied 5s metal orbital with the occupied ligand orbitals reduces the reactivity of the central atom and thus stabilizes this species. An In(III) species, [{2,4 (t)Bu(2)C(6)H(3)N=CPh}(2)(NC(5)H(3))]InCl(2)(+)InCl(4)(-) was also isolated and structurally characterized utilizing this ligand frame. PMID- 19296582 TI - Mesityl alkyne substituents for control of regiochemistry and reversibility in zirconocene couplings: new synthetic strategies for unsymmetrical zirconacyclopentadienes and conjugated polymers. AB - Reaction of 2 equivs of MesC[triple bond]CPh with Cp(2)Zr(eta(2)-Me(3)SiC[triple bond]CSiMe(3))(pyr) afforded the zirconacyclopentadiene Cp(2)Zr[2,5-Ph(2)-3,4 Mes(2)C(4)]. The regiochemistry of this isomer (betabeta with respect to the mesityl substituents) was determined through single-crystal X-ray analysis and 2D (NOESY, HSQC, HMBC) NMR experiments. This selectivity is attributed largely to a steric-based directing effect of the o-methyl ring substituents since coupling of 1,3-dimethyl-2-(phenylethynyl)benzene with zirconocene gave a single regioisomer (o-xylyl groups in both beta-positions) while coupling of 1,3-dimethyl-5 (phenylethynl)benzene gave a statistical distribution of zirconacyclopentadiene regioisomers. The coupling reaction of 2 equivs of MeC[triple bond]CMes or PrC[triple bond]CMes with Cp(2)Zr(eta(2)-Me(3)SiC[triple bond]CSiMe(3))(pyr) at ambient temperature gave the betabeta regioisomers, Cp(2)Zr[2,5-Me(2)-3,4 Mes(2)C(4)] and Cp(2)Zr[2,5-Pr(2)-3,4-Mes(2)C(4)], respectively, as the major products. Heating solutions of these zirconacycles at 80 degrees C for several hours resulted in an increase in the amount of the unsymmetrical product. For reaction mixtures of PrC[triple bond]CMes and Cp(2)Zr(eta(2)-Me(3)SiC[triple bond]CSiMe(3))(pyr) the major (and apparently thermodynamic) product under these reaction conditions was Cp(2)Zr[2,4-Mes(2)-3,5-Pr(2)C(4)]. The steric strain in the mesityl-substituted zirconacycles allowed for facile substitution reactions of MesC[triple bond]CPh or PrC[triple bond]CMes by less bulky alkynes (i.e., tolan and 3-hexyne) to give the unsymmetrical ziconacyclopentadienes Cp(2)Zr[2,4,5-Ph(3)-3-MesC(4)], Cp(2)Zr[2-Ph-3-Mes-4,5-Et(2)C(4)], and Cp(2)Zr[2 Pr-3-Mes-4,5-Ph(2)C(4)]. Reaction of a mesityl-terminated diyne containing a rigid dihexylfluorenylene spacer with zirconocene afforded poly(p fluorenylenedienylene) after demetalation with benzoic acid. PMID- 19296583 TI - Morpholino monolayers: preparation and label-free DNA analysis by surface hybridization. AB - Surface hybridization, a reaction in which nucleic acid molecules in solution react with nucleic acid partners immobilized on a surface, is widely practiced in life science research. In these applications the immobilized partner, or "probe", is typically single-stranded DNA. Because DNA is strongly charged, high salt conditions are required to enable binding between analyte nucleic acids ("targets") in solution and the DNA probes. High salt, however, compromises prospects for label-free monitoring or control of the hybridization reaction through surface electric fields; it also stabilizes secondary structure in target species that can interfere with probe-target recognition. In this work, initial steps toward addressing these challenges are taken by introducing morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of morpholino probes on gold supports can be fabricated with methods similar to those employed with DNA and are shown to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Interestingly, positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a nearly linear relationship and demonstrates sensitivities (limits of quantification) in the picogram per square millimeter range. PMID- 19296584 TI - Rh(2)(II)-catalyzed synthesis of carbazoles from biaryl azides. AB - An array of carbazoles (23 examples) can be synthesized from substituted biaryl azides at 60 degrees C using substoichiometric quantities of Rh(2)(O(2)CC(3)F(7))(4) or Rh(2)(O(2)CC(7)H(15))(4). PMID- 19296585 TI - Bistability of magnetization without spin-transition in a high-spin cobalt(II) complex due to angular momentum quenching. AB - [Co(NO(3))(2)L] (L: 2,6-di(pyrazol-1-yl)pyrazine) (1) exhibits an abrupt transition with hysteresis in magnetic susceptibility between 228 and 240 K. The results of spectroscopic and XRD measurements showed that 1 is in the high spin state in the whole temperature range. Therefore the observed hysteresis is not due to a spin transition but corresponds to a partial quenching of the angular momentum contribution to the magnetic susceptibility. Crystallographic measurements on the low- and high-temperature form of 1, combined with DFT calculations, showed that a symmetric twisting of the coordinating nitrate ions upon the transition is the most important factor in the orbital quenching mechanism. Utilizing such quenching to control magnetic properties can be a new approach to engineer transition metal complexes with magnetic functionalities without changing their spin or oxidation state. PMID- 19296586 TI - Chiral vinylphosphonate and phosphonate analogues of the immunosuppressive agent FTY720. AB - The first enantioselective synthesis of chiral isosteric phosphonate analogues of FTY720 is described. One of these analogues, FTY720-(E)-vinylphosphonate (S)-5, but not its R enantiomer, elicited a potent antiapoptotic effect in intestinal epithelial cells, suggesting that it exerts its action via the enantioselective activation of a receptor. (S)-5 failed to activate the sphingosine 1-phosphate type 1 (S1P(1)) receptor. PMID- 19296587 TI - Electrochemiluminescence of water-soluble carbon nanocrystals released electrochemically from graphite. AB - Water-soluble carbon nanocrystals (CNCs) with electrochemiluminescence (ECL) activity were released into aqueous solution from a graphite rod by applying a scanning potential. ECL emission of CNCs observed during their preparation probably provides a useful method for monitoring and screening nanocrystal preparation. The ECL behavior and its mechanism in CNCs have been studied in detail for the first time. The results suggest promising applications of CNCs in the development of new types of biosensors and display devices in the future on the basis of their strong and stable ECL emission, good stability, low cytotoxicity, excellent water solubility, easy labeling, and environmental friendliness. PMID- 19296588 TI - Gelation behavior of 2H-chromene N-acylamino acid conjugates. AB - 2H-Chromene-based conjugates of N-acyl-1,omega-amino acids (5, 9a-f, 14a-f) of natural amino acids (10a,b) and of dipeptide (10c) are prepared (60-97%) by N acylbenzotriazole methodology in aqueous media at 20 degrees C. Gelation properties of the corresponding sodium salts in DMF and DMSO are generalized with respect to an increase or decrease in the chain length of the spacer. PMID- 19296589 TI - Protonation of the side group in beta- and gamma-aminated proline analogues: effects on the conformational preferences. AB - This work shows the influence of the side-chain protonation on the conformational properties, relative stabilities, and peptide bond isomerization of four aminoproline isomers. Thus, this research has been useful to define the rules that allow control the conformation of aminoproline with the pH. Comparison of the results obtained using density functional theory calculations for the N acetyl-N'-methylamide derivatives of the protonated isomers, which differ in the beta- or gamma-position of the substituent and its cis or trans relative disposition, with those reported for the corresponding neutral analogues (J. Phys. Chem. B 2008, 112, 14045) has allowed us to reach the following conclusions: (i) protonation of the amino group produces a reduction of the backbone conformational flexibility and a destabilization of the cis configuration of the amide bond involving the pyrrolidine nitrogen; (ii) the planarity of the peptide bond is broken in some cases to form strong side chain...backbone interactions, which induce a very significant pyramidilization at the amide nitrogen atom; (iii) as was also detected for the neutral analogues, the formation of side chain...backbone intraresidue interactions favor the cis disposition of the substituent; and (iv) protonation of the amino side group increases the energy gaps that separate the different investigated isomers resulting in an enhancement of the destabilization of the dipeptides with the substituent attached in a trans position. PMID- 19296590 TI - Facile ionothermal synthesis of microporous and mesoporous carbons from task specific ionic liquids. AB - An expedient, template-free, high-yield, and solventless route to nitrogen-rich micro- and mesoporous carbons is reported based on direct, atmospheric-pressure carbonization of task-specific ionic liquids bearing one or more nitrile side chains. The resulting textural properties (pore regime, surface area) are highly dependent upon the structural motifs of the ions comprising the corresponding parent ionic liquid, and uniform carbon films are routinely deposited with this novel methodology, highlighting excited new opportunities in the development of advanced functional carbon composites. PMID- 19296591 TI - High-throughput pesticide residue quantitative analysis achieved by tandem mass spectrometry with automated flow injection. AB - The use of automated flow injection with MS/MS detection for fast quantitation of agrochemicals in food and water samples was demonstrated in this study. Active ingredients from the sulfonylurea herbicide and carbamate insecticide classes were selected as model systems. Samples were prepared using typical procedures from residue methods, placed in an autosampler, and injected directly into a triple quadrupole instrument without chromatographic separation. The technique allows data acquisition in 15 s per injection, with samples being injected every 65 s, representing a significant improvement from the 15-30 min needed in typical HPLC/MS/MS methods. The availability of HPLC systems is an advantage since they can be used in flow-injection mode (bypassing the column compartment). Adequate accuracy, linearity, and precision (R(2) > 0.99 and RSD < 20%) were obtained using external standards prepared in each control matrix. The limit of quantitation (LOQ) achieved for all analytes was 0.01 mg/kg in food samples and 0.1 ng/mL in water; while limits of detection (LOD) were estimated to be about 0.003 mg/kg and 0.03 ng/mL in food and water, respectively. The advantages and limitations of flow injection MS/MS for ultratrace-level quantitative analysis in complex matrixes are discussed. PMID- 19296592 TI - Gel electrophoresis method to measure the concentration of single-walled carbon nanotubes extracted from biological tissue. AB - A rapid and sensitive method to detect single-walled carbon nanotubes (SWNTs) in biological samples is presented. The method uses polyacrylamide gel electrophoresis (PAGE) followed by quantification of SWNT bands. SWNTs dispersed in bovine serum albumin (BSA) were used to develop the method. When BSA-SWNT dispersions were subjected to sodium dodecyl sulfate (SDS)-PAGE, BSA passed through the stacking gel, entered the resolving gel, and migrated toward the anode as expected. The SWNTs, however, accumulated in a sharp band at the interface between the loading well and the stacking gel. The intensities from digitized images of these bands were proportional to the amount of SWNTs loaded onto the gel with a detection limit of 5 ng of SWNTs. To test the method, normal rat kidney (NRK) cells in culture were allowed to take up SWNTs upon exposure to medium containing various concentrations of BSA-SWNTs for different times and temperatures. The SDS-PAGE analyses of cell lysate samples suggest that BSA-SWNTs enter NRK cells by fluid-phase endocytosis at a rate of 30 fg/day/cell upon exposure to medium containing 98 microg/mL SWNTs. PMID- 19296593 TI - Alpha-alkenoyl ketene S,S-acetal-based multicomponent reaction: an efficient approach for the selective construction of polyfunctionalized cyclohexanones. AB - A versatile multicomponent reaction based on the new four-carbon synthons alpha alkenoyl ketene S,S-acetals 1 has been developed. This three-component reaction of readily available alpha-alkenoyl ketene S,S-acetals 1 with aldehydes 2 and active methylene compounds 3 proceeds smoothly in acidic medium (glacial acetic acid in tetrahydrofuran) to give various polyfunctionalized cyclohexanones 4, 5, and 6 in a highly regio- and diastereoselective manner with good to excellent yields. The reaction can tolerate a broad range of substituents in the three components involved and is proposed to proceed via a tandem Knoevenagel intermolecular Michael-intramolecular Michael sequence. As an extension of the synthetic application of polyfunctionalized cyclohexanones obtained, unsymmetrical biaryls 7 were synthesized in almost quantitative yields by simple transformations of the corresponding cycloadducts 6. PMID- 19296594 TI - Microsystem for field-amplified electrokinetic trapping preconcentration of DNA at poly(ethylene terephthalate) membranes. AB - In electrokinetic trapping (EKT), the electroosmotic velocity of a buffer solution in one area of a microfluidic device opposes the electrophoretic velocity of the analyte in a second area, resulting in transport of DNA to a location where the electrophoretic and electroosmotic velocities are equal and opposite and DNA concentrates at charged nanochannels. The method does not require an optical plug localization, a considerable advantage as compared to preconcentration techniques previously presented. In the work reported here, the trapping process is preceded by a field-amplification in the sample reservoir to reduce trapping time, as field-amplified EKT is shown to be an effective technique to preconcentrate samples from larger volumes. A theoretical model explaining the principle of field-amplified EKT considers different ionic strengths and cross-sectional areas in the microchip segments. The model is supported by experimental data using nucleic acids and fluorescein as sample analytes. An incorporated poly(ethylene terephthalate) (PET) membrane provides anion exclusion due to a negatively charged surface. A fluidic counter flow supports the trapping process in 100 nm pores due to anion exclusion. An analysis of Joule heating gives evidence that temperature gradient focusing effects are negligible and charge exclusion is responsible for trapping. The theoretical model developed and experimentally demonstrated can be exploited for the preconcentration of cell free fetal DNA circulating in maternal plasma and other rare nucleic acid species present in large sample volumes. PMID- 19296595 TI - Spectroscopic identification of tri-n-octylphosphine oxide (TOPO) impurities and elucidation of their roles in cadmium selenide quantum-wire growth. AB - Tri-n-octylphosphine oxide (TOPO) is the most commonly used solvent for the synthesis of colloidal nanocrystals. Here we show that the use of different batches of commercially obtained TOPO solvent introduces significant variability into the outcomes of CdSe quantum-wire syntheses. This irreproducibility is attributed to varying amounts of phosphorus-containing impurities in the different TOPO batches. We employ (31)P NMR to identify 10 of the common TOPO impurities. Their beneficial, harmful, or negligible effects on quantum-wire growth are determined. The impurity di-n-octylphosphinic acid (DOPA) is found to be the important beneficial TOPO impurity for the reproducible growth of high quality CdSe quantum wires. DOPA is shown to beneficially modify precursor reactivity through ligand substitution. The other significant TOPO impurities are ranked according to their abilities to similarly influence precursor reactivity. The results are likely of general relevance to most nanocrystal syntheses conducted in TOPO. PMID- 19296596 TI - Chemoenzymatic synthesis of a new class of macrocyclic oligosaccharides. AB - A novel and highly efficient chemoenzymatic method has been developed for the preparation of structurally defined macrocyclic oligosaccharides of varied sizes. This method involves chemical or chemoenzymatic synthesis of oligosaccharides containing a galactose at the nonreducing end and a propargyl group at the reducing end as sialyltransferase acceptors. Introducing an azido-containing sialic acid to the nonreducing end of the galactosides through a sialyltransferase-catalyzed enzymatic reaction followed by copper(I)-catalyzed Huisgen's 1,3-dipolar cycloaddition of alkyne and azide provides size-defined macrocyclic carbohydrates. The produced negatively charged macrocycles have high solubility in water and interact with hydrophobic small molecules in a size dependent manner. PMID- 19296597 TI - N(epsilon)-thioacetyl-lysine-containing tri-, tetra-, and pentapeptides as SIRT1 and SIRT2 inhibitors. AB - N()-Thioacetyl-lysine-containing tri-, tetra-, and pentapeptides, based on the alpha-tubulin and p53 protein sequences, were studied as SIRT1 and SIRT2 inhibitors. The potency of the pentapeptides depended on the selection of the side chains. The removal of N- and C-terminal residues of the pentapeptides yielded tripeptides with retained SIRT1 inhibitory activity but decreased SIRT2 inhibitory activity. The most potent SIRT1 inhibitors were equipotent with the reference compound (6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide) with the IC(50) values of 180-330 nM. PMID- 19296598 TI - Identification of novel antibacterial peptides by chemoinformatics and machine learning. AB - The rise of antibiotic resistant pathogens is one of the most pressing global health issues. Discovery of new classes of antibiotics has not kept pace; new agents often suffer from cross-resistance to existing agents of similar structure. Short, cationic peptides with antimicrobial activity are essential to the host defenses of many organisms and represent a promising new class of antimicrobials. This paper reports the successful in silico screening for potent antibiotic peptides using a combination of QSAR and machine learning techniques. On the basis of initial high-throughput measurements of activity of over 1400 random peptides, artificial neural network models were built using QSAR descriptors and subsequently used to screen an in silico library of approximately 100,000 peptides. In vitro validation of the modeling showed 94% accuracy in identifying highly active peptides. The best peptides identified through screening were found to have activities comparable or superior to those of four conventional antibiotics and superior to the peptide most advanced in clinical development against a broad array of multiresistant human pathogens. PMID- 19296599 TI - Substituted terphenyl compounds as the first class of low molecular weight allosteric inhibitors of the luteinizing hormone receptor. AB - The luteinizing hormone (LH) receptor plays an important role in fertility and certain cancers. The endogenous ligands human chorionic gonadotropin (hCG) and LH bind to the large N terminal domain of the receptor. We recently reported on the first radiolabeled low molecular weight (LMW) agonist for this receptor, [(3)H]Org 43553, which was now used to screen for new LMW ligands. We identified a terphenyl derivative that inhibited [(3)H]Org 43553 binding to the receptor, which led us to synthesize a number of derivatives. The most potent compound of this terphenyl series, 24 (LUF5771), was able to increase the dissociation rate of [(3)H]Org 43553 by 3.3-fold (at 10 muM). In a functional assay, the presence of 24 resulted in a 2- to 3-fold lower potency of both Org 43553 and LH. Thus, the compounds presented in this paper are the first LMW ligands that allosterically inhibit the LH receptor. PMID- 19296600 TI - Novel peptidomimetics containing a vinyl ester moiety as highly potent and selective falcipain-2 inhibitors. AB - This paper describes the synthesis and biological evaluation of a new class of peptidomimetic falcipain-2 inhibitors based on a 1,4-benzodiazepine scaffold combined with various alpha,beta-unsaturated electrophilic functions such as vinyl-ketone, -amide, -ester, and -nitrile. The profile of reactivity of this class of derivatives has been evaluated and 4c, containing a vinyl ester warhead, proved to be a highly potent and selective falcipain-2 inhibitor. PMID- 19296601 TI - Facile synthesis of novel two- and three-dimensional coordination polymers containing dialkyltin phosphonate-based tri/tetra-nuclear clusters with appended sulfonate groups. AB - The coordination-driven self-assemblies of mixed-ligand dialkyltin derivatives, [(Et(2)Sn)(4) (O(2)P(OH)Me)(2)(O(3)PMe)(2)(OSO(2)Et)(2) x 2 H(2)O](n) 1, [(Et(2)Sn)(3)(O(3)PMe)(2)(OSO(2)Me)(2) x CHCl(3)](n) 2, and [(Me(2)Sn)(3)(O(3)PBu(t))(2)(OSO(2)Me)(2) x 2 CHCl(3)](n) 3 have been achieved by reacting the tin precursors, [R(2)Sn(OR(1))(OSO(2)R(1))](n) (R = Et, R(1) = Et (1a), Me (2a); R = Me, R(1) = Me (3a)) with an equimolar amount of methylphosphonic/t-butylphosphonic acid under mild conditions (rt, 8 h, CH(2)Cl(2)). These have been characterized by IR and multinuclear ((1)H, (13)C, (31)P, and (119)Sn) NMR spectroscopy as well as single crystal X-ray diffraction. The asymmetric unit of 1 is composed of a tetranuclear, Sn(4)(mu(2) PO(2))(2)(mu(3)-PO(3))(2) core bearing an appended ethanesulfonate group on each terminal tin (Sn2) atom and two P(OH)...O hydrogen bonded water molecules. The ladder-like structural motif thus formed is extended into one-dimensional polymeric chains by virtue of bridging bidentate mode of the sulfonate groups. These chains are linked by O-H...O(S) hydrogen bonds involving H(2)O molecules and oxygen atoms of the sulfonate groups. The asymmetric units of 2 and 3 are composed of trinuclear tin clusters with a Sn(3)(mu(3)-PO(3))(2) core and two dangling methanesulfonate groups which are covalently bonded to the tin centers. The construction of three-dimensional self-assemblies is effected by variable bonding modes (mu(2), mu(3) in 2; mu(2) in 3) of the methanesulfonate groups. Both the structural motifs possess five- and six-coordinated tin atoms and form rectangular channels which are occupied by CHCl(3) molecules. PMID- 19296602 TI - Group 11 complexes of the P,Te-centered ligand [TeP(i)Pr2NP(i)Pr2]-: synthesis, structures, and insertion reactions of the copper(I) complex with chalcogens. AB - The lithium reagent [LiTeP(i)Pr(2)NP(i)Pr(2)] undergoes metathetical reactions with group 11 chlorides to give the complexes {M(TeP(i)Pr(2)NP(i)Pr(2))}(3) (6: M = Cu; 7: M = Ag) and (Ph(3)P)Au(TeP(i)Pr(2)NP(i)Pr(2)) (8) as yellow crystalline solids. These new complexes were characterized by single crystal X-ray diffraction and multinuclear NMR spectroscopy. The tellurium atoms in the trimeric complexes 6 and 7 occupy bridging positions to give a Cu(3)Te(3) ring in a twist-boat conformation with short M...M contacts (M = Cu, Ag). Variable temperature (31)P NMR and ESI-MS spectra for 6 and 7 give evidence for the formation of several oligomers in tetrahydrofuran solution. The reactions of 6 with dioxygen (or Me(3)NO), elemental sulfur, or red selenium generate the chalcogen-insertion products {Cu(TeP(i)Pr(2)NP(i)Pr(2)E)}(3) (9: E = O; 10: E = S; 11: E = Se), which were also structurally characterized by single crystal X ray diffraction and multinuclear NMR spectroscopy. The lighter chalcogens are two coordinate while the tellurium centers occupy bridging positions in the trimeric complexes 9-11 giving rise to Cu(3)Te(3) rings in a chairlike conformation. PMID- 19296603 TI - Ligand-dependent ultrasonic-assistant self-assemblies and photophysical properties of lanthanide nicotinic/isonicotinic complexes. AB - Two structural series, including two isomorphous homodinuclear complexes Ln(2)(H(2)O)(4)(C(6)NO(2)H(4))(6) (Ln = Tb (1) and Er (2)) and four isostructural one-dimensional (1-D) chain-like assemblies [Ln(H(2)O)(4)(C(6)NO(2)H(4))(2)](n) x nCl (Ln = Sm (3), Eu (4), Tb (5), and Dy (6)), have been rationally prepared through a facile ultrasonic synthesis and have been characterized by X-ray diffraction and photophysical measurements. Both complexes, 1 and 2, feature a homodinuclear structure, based on two 8-fold coordination lanthanide atoms bridged by four nicotinic acid ligands. Complexes 3-6 are characterized by a 1-D polycationic chain-like structure, containing eight-coordinated lanthanide ions and bridging isonicotinic acid ligands. The 1-D polycationic chains and the isolated chloride anions are interconnected via hydrogen bonds and pi-pi interactions to form a three-dimensional supramolecular network. The effect of nicotinic/isonicotinic acid ligands on the structures and the photoluminescence properties, as well as the relationship between the photoluminescence properties and the structures, was investigated based on IR, UV-vis absorption spectra, low temperature phosphorescent spectra, excitation, and emission spectra. The fluorescence quantum yields of complexes 1 and 2 were determined to be 44% and 21%, respectively. PMID- 19296604 TI - Synthesis and characterization of mixed chalcogen triangular complexes with new Mo3(mu3-S)(mu2-Se2)3(4+) and M3(mu3-S)(mu2-Se)3(4+) (M = Mo, W) cluster cores. AB - In our pursuit of mixed chalcogen-bridged cluster complexes, solids of the compositions Mo(3)SSe(6)Br(4) and W(3)SSe(6)Br(4) were prepared using high temperature synthesis from the elements. Treatment of Mo(3)SSe(6)Br(4) with Bu(4)NBr in a vibration mill yielded (Bu(4)N)(3){[Mo(3)(mu(3)-S)(mu(2) Se(2))(3)Br(6)]Br} (I). Its all-selenide analogue (Bu(4)N)(3){[Mo(3)(mu(3) Se)(mu(2)-Se(2))(3)Br(6)]Br} (II) was prepared from Mo(3)Se(7)Br(4) in a similar way. Both compounds were characterized by IR, Raman, and (77)Se NMR spectroscopy. The structure of II was determined by X-ray single-crystal analysis. Compound I is isostructural with II and contains the new Mo(3)(mu(3)-S)Se(6)(4+) cluster core. By treatment of a 4 M Hpts solution of I with PPh(3) followed by cation exchange chromatography, the new mixed chalcogenido-molybdenum aqua ion, [Mo(3)(mu(3)-S)(mu(2)-Se)(3)(H(2)O)(9)](4+), was isolated and characterized using UV-vis spectroscopy and, after derivatization into [Mo(3)(mu(3)-S)(mu(2) Se)(3)(acac)(3)(py)(3)](+), electrospray ionization mass spectrometry. From HCl solutions of the aqua ion, a supramolecular adduct with cucurbit[6]uril (CB[6]), {[Mo(3)(mu(3)-S)(mu(2)-Se)(3)(H(2)O)(6)Cl(3)](2)CB[6]}Cl(2) x 11 H(2)O (III), was isolated and its structure determined using X-ray crystallography. W(3)SSe(6)Br(4) upon reaction with H(3)PO(2) gave a mixture of all of the [W(3)S(x)Se(4-x)(H(2)O)(9)](4+) species. After repeated chromatography, crystals of {[W(3)(mu(3)-S)(mu(2)-Se)(3)(H(2)O)(7)Cl(2)](2)CB[6]}Cl(4) x 12 H(2)O (IV) were crystallized from the fraction rich in [W(3)(mu(3)-S)Se(3)(H(2)O)(9)](4+) and structurally characterized. PMID- 19296605 TI - Development of an oxime bond containing daunorubicin-gonadotropin-releasing hormone-III conjugate as a potential anticancer drug. AB - Here, we report on the synthesis and biological properties of a conjugate in which daunorubicin (Dau) as chemotherapeutic agent was attached through an oxime bond to gonadotropin-releasing hormone-III (GnRH-III) as targeting moiety. In vitro toxicity and the cytostatic effect of the conjugate on MCF-7 human breast and C26 murine colon cancer cell lines were determined, and the results were compared with those obtained for the free daunorubicin, as well as with the doxorubicin containing derivative. In vivo antitumor effect of daunorubicin-GnRH III was studied on Balb/c female mice transplanted with C26 tumor. Our data indicate that the daunorubicin-GnRH-III conjugate had a lower toxic effect than the free daunorubicin and it was essentially nontoxic up to 15 mg (Dau content)/kg body weight. The treatment of the C26 tumor bearing mice with the conjugate led to tumor growth inhibition and longer survival time in comparison with the controls and with the administration of the free drug. When mice were treated twice with the conjugate (on days 4 and 7 after tumor transplantation), 46% tumor growth inhibition was obtained. In this case, the increase of the median survival time was 38% compared to the controls. PMID- 19296606 TI - Electrical manipulation of nanofilaments in transition-metal oxides for resistance-based memory. AB - The fabrication of controlled nanostructures such as quantum dots, nanotubes, nanowires, and nanopillars has progressed rapidly over the past 10 years. However, both bottom-up and top-down methods to integrate the nanostructures are met with several challenges. For practical applications with the high level of the integration, an approach that can fabricate the required structures locally is desirable. In addition, the electrical signal to construct and control the nanostructures can provide significant advantages toward the stability and ordering. Through experiments on the negative resistance switching phenomenon in Pt-NiO-Pt structures, we have fabricated nanofilament channels that can be electrically connected or disconnected. Various analyses indicate that the nanofilaments are made of nickel and are formed at the grain boundaries. The scaling behaviors of the nickel nanofilaments were closely examined, with respect to the switching time, power, and resistance. In particular, the 100 nm x 100 nm cell was switchable on the nanosecond scale, making them ideal for the basis for high-speed, high-density, nonvolatile memory applications. PMID- 19296607 TI - Accurate theoretical chemistry with coupled pair models. AB - Quantum chemistry has found its way into the everyday work of many experimental chemists. Calculations can predict the outcome of chemical reactions, afford insight into reaction mechanisms, and be used to interpret structure and bonding in molecules. Thus, contemporary theory offers tremendous opportunities in experimental chemical research. However, even with present-day computers and algorithms, we cannot solve the many particle Schrodinger equation exactly; inevitably some error is introduced in approximating the solutions of this equation. Thus, the accuracy of quantum chemical calculations is of critical importance. The affordable accuracy depends on molecular size and particularly on the total number of atoms: for orientation, ethanol has 9 atoms, aspirin 21 atoms, morphine 40 atoms, sildenafil 63 atoms, paclitaxel 113 atoms, insulin nearly 800 atoms, and quaternary hemoglobin almost 12,000 atoms. Currently, molecules with up to approximately 10 atoms can be very accurately studied by coupled cluster (CC) theory, approximately 100 atoms with second-order Moller Plesset perturbation theory (MP2), approximately 1000 atoms with density functional theory (DFT), and beyond that number with semiempirical quantum chemistry and force-field methods. The overwhelming majority of present-day calculations in the 100-atom range use DFT. Although these methods have been very successful in quantum chemistry, they do not offer a well-defined hierarchy of calculations that allows one to systematically converge to the correct answer. Recently a number of rather spectacular failures of DFT methods have been found even for seemingly simple systems such as hydrocarbons, fueling renewed interest in wave function-based methods that incorporate the relevant physics of electron correlation in a more systematic way. Thus, it would be highly desirable to fill the gap between 10 and 100 atoms with highly correlated ab initio methods. We have found that one of the earliest (and now almost forgotten) of this class of methods, the coupled-electron pair approximation (CEPA), performs exceedingly well in chemical applications. In this Account, we examine the performance of CEPA in chemical applications. One attractive feature of CEPA, in addition to its surprising accuracy that surpasses that of DFT and MP2 theory, is a simplicity that allows for straightforward and very efficient approximations and extensions to be developed; these are much more difficult or even impossible with the more rigorous CC theory. Thus, approximate CEPA methods can be implemented efficiently enough to allow for calculations on molecules of 50-100 atoms, perhaps the most common range in contemporary chemical research. PMID- 19296608 TI - Properties of one-dimensional molybdenum nanowires in a confined environment. AB - The atomistic mechanism for the self-assembly of molybdenum into one-dimensional metallic nanowires in a confined environment such as a carbon nanotube is investigated using quantum mechanical calculations. We find that Mo does not organize into linear chains but rather prefers to form four atom per unit cell nanowires that consist of a subunit of a Mo body-centered cubic crystal. Our model explains the 0.3 nm separation between features measured by high-resolution transmission electron microscopy and why the nanotube diameter must be in the 0.70-1.0 nm range to accommodate the smallest stable one-dimensional wire. We also computed the electronic band structure of the Mo wires inside a nanotube and found significant hybridization with the nanotube states, thereby explaining the experimentally observed quenching of fluorescence and the damping of the radial breathing modes as well as an increased resistance to oxidation. PMID- 19296609 TI - Facile cycling of Ti-doped LiAlH4 for high performance hydrogen storage. AB - LiH and Ti-doped Al react quantitatively with H(2) in Me(2)O solution to form LiAlH(4) under mild conditions. The solvent is easily vented along with excess H(2) on completion, leaving dry Ti-doped LiAlH(4); this releases approximately 7 wt % H(2) commencing at 80 degrees C with excellent kinetics. PMID- 19296610 TI - Vertical epitaxial wire-on-wire growth of Ge/Si on Si(100) substrate. AB - Vertically aligned epitaxial Ge/Si heterostructure nanowire arrays on Si(100) substrates were prepared by a two-step chemical vapor deposition method in anodic aluminum oxide templates. n-Butylgermane vapor was employed as new safer precursor for Ge nanowire growth instead of germane. First a Si nanowire was grown by the vapor liquid solid growth mechanism using Au as catalyst and silane. The second step was the growth of Ge nanowires on top of the Si nanowires. The method presented will allow preparing epitaxially grown vertical heterostructure nanowires consisting of multiple materials on an arbitrary substrate avoiding undesired lateral growth. PMID- 19296611 TI - Characterizing loop dynamics and ligand recognition in human- and avian-type influenza neuraminidases via generalized born molecular dynamics and end-point free energy calculations. AB - The comparative dynamics and inhibitor binding free energies of group-1 and group 2 pathogenic influenza A subtype neuraminidase (NA) enzymes are of fundamental biological interest and relevant to structure-based drug design studies for antiviral compounds. In this work, we present seven generalized Born molecular dynamics simulations of avian (N1)- and human (N9)-type NAs in order to probe the comparative flexibility of the two subtypes, both with and without the inhibitor oseltamivir bound. The enhanced sampling obtained through the implicit solvent treatment suggests several provocative insights into the dynamics of the two subtypes, including that the group-2 enzymes may exhibit similar motion in the 430-binding site regions but different 150-loop motion. End-point free energy calculations elucidate the contributions to inhibitor binding free energies and suggest that entropic considerations cannot be neglected when comparing across the subtypes. We anticipate the findings presented here will have broad implications for the development of novel antiviral compounds against both seasonal and pandemic influenza strains. PMID- 19296612 TI - Multimetallic arrays: bi-, tri-, tetra-, and hexametallic complexes based on gold(I) and gold(III) and the surface functionalization of gold nanoparticles with transition metals. AB - Reaction of [AuCl(PPh(3))] with the zwitterion S(2)CNC(4)H(8)NH(2) yields [(Ph(3)P)Au(S(2)CNC(4)H(8)NH(2))]BF(4). Treatment of this species with NEt(3) and CS(2) followed by [AuCl(PPh(3))] leads to [{(Ph(3)P)Au}(2)(S(2)CNC(4)H(8)NCS(2))], which can also be obtained directly from [AuCl(PPh(3))] and KS(2)CNC(4)H(8)NCS(2)K. A heterobimetallic variant, [(dppm)(2)Ru(S(2)CNC(4)H(8)NCS(2))Au(PPh(3))](+), can be prepared by the sequential reaction of [(dppm)(2)Ru(S(2)CNC(4)H(8)NH(2))](2+) with NEt(3) and CS(2) followed by [AuCl(PPh(3))]. Reaction of the same ruthenium precursor with [(dppm)(AuCl)(2)] under similar conditions yields the trimetallic complex [(dppm)(2)Ru(S(2)CNC(4)H(8)NCS(2))Au(2)(dppm)](2+). Attempts to prepare the compound [(dppm)Au(2)(S(2)CNC(4)H(8)NH(2))](2+) from [(dppm)(AuCl)(2)] led to isolation of the known complex [{(dppm)Au(2)}(2)(S(2)CNC(4)H(8)NCS(2))](2+) via a symmetrization pathway. [{(dppf)Au(2)}(2)(S(2)CNC(4)H(8)NCS(2))](2+) was successfully prepared from [(dppf)(AuCl)(2)] and crystallographically characterized. In addition, a gold(III) trimetallic compound, [{(dppm)(2)Ru(S(2)CNC(4)H(8)NCS(2))}(2)Au](3+), and a tetrametallic gold(I) species, [{(dppm)(2)Ru(S(2)CNC(4)H(8)NCS(2))Au}(2)](2+), were also synthesized. This methodology was further exploited to attach the zwitterionic (dppm)(2)Ru(S(2)CNC(4)H(8)NCS(2)) unit to the surface of gold nanoparticles, which were generated in situ and found to be 3.4 (+/-0.3) and 14.4 (+/-2.5) nm in diameter depending on the method employed. Nanoparticles with a mixed surface topography were also explored. PMID- 19296613 TI - Molecular tools for the self-assembly of bisporphyrin photodyads: a comprehensive physicochemical and photophysical study. AB - Accessible and hindered phenanthroline-strapped Zn(II) porphyrin receptors exhibited suitable topography tailored to strongly and selectively bind N(1) unsubstituted imidazoles and imidazoles appended to free-base porphyrins. Distal binding was clearly driven by the formation of strong bifurcated hydrogen bonds with the phenanthroline unit of the receptors. An extensive physicochemical study emphasized the influence of bulkiness of the substrate and of the porphyrin receptor on the binding and self-assembly mechanism. Knowledge of the corresponding spectroscopic, thermodynamic, and kinetic data were of fundamental importance to elucidate and to model the photoinduced properties which occur within the self-assembled porphyrin dyads. PMID- 19296615 TI - Electronics and magnetism of patterned graphene nanoroads. AB - Individual ribbons of graphene show orientation-dependent electronic properties of great interest, yet to ensure their perfect geometry and integrity or to assemble free ribbons into a device remains a daunting task. Here we explore, using density functional theory, an alternative possibility of "nanoroads" of pristine graphene being carved in the electrically insulating matrix of fully hydrogenated carbon sheet (graphane). Such one-dimensional entities show individual characteristics and, depending upon zigzag (and their magnetic state) or armchair orientation, can be metallic or semiconducting. Furthermore, the wide enough zigzag roads become magnetic with energetically similar ferro- and antiferromagnetic states. Designing magnetic, metallic, and semiconducting elements within the same mechanically intact sheet of graphene presents a new opportunity for applications. PMID- 19296614 TI - Effect of modification of the physicochemical properties of ICAM-1-derived peptides on internalization and intracellular distribution in the human leukemic cell line HL-60. AB - The objective of this work is to test the hypothesis that increasing the hydrophilicity of DOX-peptide conjugates may modify their entry mechanisms into HL-60 cells from passive diffusion to receptor-mediated uptake. To test this hypothesis, the entry mechanisms and the intracellular disposition of DOX-cIBR7, DOX-PEGcIBR7, FITC-cIBR, and FITC-cIBR7 were evaluated in HL-60 cells. To increase the hydrophilicity of the peptide, the cIBR peptide (cyclo(1,12)Pen PRGGSVLVTGC) was modified to cIBR7 peptide (cyclo(1,8)CPRGGSVC) by removing the hydrophobic residues at the C-terminus. DOX-cIBR7 conjugate, which has higher hydrophilicity than DOX-cIBR, was synthesized. Second, a hydrophilic linker (11 amino-3,6,9-trioxaundecanate linker) was incorporated between DOX and cIBR7 to generate DOX-PEGcIBR7 with higher hydrophilicity than DOX-cIBR7. As controls, FITC-cIBR and FITC-cIBR7 were used to check for any endocytic uptake process of the peptide. As previously found with DOX-cIBR, DOX-cIBR7, and DOX-PEGcIBR7, conjugates enter the cells via passive diffusion and not via the energy-dependent endocytic process. This result suggests that an increase in hydrophilicity does not influence the entry mechanism of the DOX-peptide conjugates. In contrast to the DOX-cIBR7 conjugate, the FITC-cIBR7 conjugate showed energy-dependent cellular entry into the cells and followed an endocytic pathway similar to that for dextran. Finally, the entry of DOX-cIBR7 and DOX-PEGcIBR into the cell cytosol was shown to be due to the properties of DOX and not to those of the peptide. PMID- 19296616 TI - Cytotoxic and antiplasmodial xanthones from Pentadesma butyracea. AB - Four new xanthones, butyraxanthones A-D (1-4), were isolated from the stem bark of Pentadesma butyracea, together with six known xanthones (5-10) and a triterpenoid (lupeol). The structures of 1-4 were established by spectroscopic methods. Compounds 1-10 were tested in vitro for antiplasmodial activity against a Plasmodium falciparum chloroquine-resistant strain and for cytotoxicity against a human breast cancer cell line (MCF-7). Nearly all of these xanthones exhibited good antiplasmodial activity, and some of them also demonstrated potent cytotoxicity. PMID- 19296617 TI - Antioxidant Lignans from the Fruits of Broussonetia papyrifera. AB - Nine new lignans, chushizisins A-I (1-9), and three known lignans, threo-1-(4 hydroxy-3-methoxyphenyl)-2-{4-[(E)-3-hydroxy-1-propenyl]-2-methoxyphenoxy}-1,3 propanediol (10), erythro-1-(4-hydroxy-3-methoxyphenyl)-2-{4-[(E)-3-hydroxy-1 propenyl]-2-methoxyphenoxy}-1,3-propanediol (11), and 3-[2-(4- hydroxyphenyl)-3 hydroxymethyl-2,3-dihydro-1-benzofuran-5-yl]propan-1-ol (12), were isolated from the fruits of Broussonetia papyrifera. Their structures were elucidated using spectroscopic methods. Compounds 1, 5, 6, 8, 9, and 11 exhibited antioxidant activities against H(2)O(2)-induced impairment in PC12 cells, while compounds 1, 2, 4, 7, and 11 showed DPPH radical-scavenging activities with IC(50) values of 236.8, 156.3, 273.9, 281.1, and 60.9 microM, respectively. PMID- 19296618 TI - Synthesis, crystal structure, and physical properties of 7,14-disubstituted pentacene-5,12-diones containing a methylenequinoid structure. AB - Title 7,14-disubsituted pentacene-5,12-diones were prepared for the first time by a nucleophilic substitution reaction of a pentacene-5,7,12,14-tetraone with aryl- or ethynyllithiums followed by a dehydroxylation reaction. The methylenequinoid structures with two conjugated carbonyl groups were clearly observed in their crystal structures. They showed intense absorptions in the visible region and amphoteric redox properties with high reduction potentials. PMID- 19296619 TI - Rational selection of gold nanorod geometry for label-free plasmonic biosensors. AB - We present the development of an analytical model that can be used for the rational design of a biosensor based on shifts in the local surface plasmon resonance (LSPR) of individual gold nanoparticles. The model relates the peak wavelength of light scattered by an individual plasmonic nanoparticle to the number of bound analyte molecules and provides an analytical formulation that predicts relevant figures-of-merit of the sensor such as the molecular detection limit (MDL) and dynamic range as a function of nanoparticle geometry and detection system parameters. The model calculates LSPR shifts for individual molecules bound by a nanorod, so that the MDL is defined as the smallest number of bound molecules that is measurable by the system, and the dynamic range is defined as the maximum number of molecules that can be detected by a single nanorod. This model is useful because it will allow a priori design of an LSPR sensor with figures-of-merit that can be optimized for the target analyte. This model was used to design an LSPR sensor based on biotin-functionalized gold nanorods that offers the lowest MDL for this class of sensors. The model predicts a MDL of 18 streptavidin molecules for this sensor, which is in good agreement with experiments and estimates. Further, we discuss how the model can be utilized to guide the development of future generations of LSPR biosensors. PMID- 19296620 TI - Rectifying and sorting of regenerating axons by free-standing nanowire patterns: a highway for nerve fibers. AB - We present an EBL-defined nanowire pattern that can sort axons coming from different directions on a substrate. The pattern defines tracks for left-bound traffic and right-bound traffic, which opens up new possibilities for designing neural networks on a chip. PMID- 19296621 TI - Fast and robust route to hydroporphyrin-chalcones with extended red or near infrared absorption. AB - The reaction of an acetylchlorin or diacetylbacteriochlorin with an aldehyde under microwave conditions readily affords the corresponding hydroporphyrin chalcone. The aldehydes include aryl aldehydes, cinnamaldehyde, and all-trans retinal. The chalcone causes a bathochromic shift of the long-wavelength absorption band of the hydroporphyrin by up to 24 nm. The facile conjugation and wavelength tunability should make such constructs valuable for fundamental spectroscopic studies as well as diverse photochemical applications in the relatively unexplored red and near-infrared spectral regions. PMID- 19296622 TI - Growth of solid domains in model membranes: quantitative image analysis reveals a strong correlation between domain shape and spatial position. AB - The nucleation and growth of solid domains in supported bilayers composed of a binary mixture of equimolar 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) have been studied using combined fluorescence microscopy and AFM. We have found that the formation of the DPPC-enriched solid domains occurs by a combination of homogeneous and heterogeneous nucleation and that the nucleation density is directly proportional to the cooling rate. Furthermore, during cooling the shape of the domains evolve from compact to a branched morphology. This suggests that the growth is controlled by the diffusion of DPPC from the liquid phase toward the solid domain interface. In the late stages of the growth, we observe that the size and overall shape of the domains depend on the position of the nucleation points relative to the surrounding nucleation point positions. To analyze this effect, the nucleation points were used as generators in a Voronoi construction. Associated with each generator is a Voronoi polygon that contains all points closer to this generator than to any other. Through a detailed quantitative analysis of the Voronoi cells and the domains, we have found that their area, orientation, and asymmetry correlate and that the correlation becomes stronger for larger domains. This means that the spatial distribution of the nucleation points regulate the domain shape. PMID- 19296623 TI - Pressure-induced crystal memory effect of spider silk proteins. AB - Spider silk is a novel material with high strength and toughness and is lightweight, making it attractive for high-performance fiber and biomedical applications. The amino acid sequence and molecular architecture of spider silk protein are becoming known gradually with science and technology development. However, its relationship of structure and function is a puzzle which attracts materials scientists to unveil it. In this study we investigated the changes of the crystal structure as a result of different processing techniques to spider silk protein. Although the crystallization of biomacromolecules is very difficult and complex, we did observe the crystal of electrospun spider silk protein treated by external high pressure in its solid state. This study also provides us a simple and effective method to prepare protein crystals. It is most important that this is the first time that we found spider silk protein as a result of electrospinning that had a crystallization memory property when it was stimulated by a high pressure, which will further extend the list of its particular properties. This finding will also lead us to study it deeply and widen our research on proteins of other organisms. PMID- 19296624 TI - Density functional approach to adsorption and retention of spherical molecules on surfaces modified with end-grafted polymers. AB - A density functional approach to describe adsorption of Lennard-Jones fluids on a surface modified with grafted chains is proposed. The theory is extended to the solute retention in chromatography with chemically bonded phases. The chain molecules are modeled as freely jointed tangent spheres with end segments linked to the surface. The segments interact via Lennard-Jones potential. The effects of grafting density and molecular interactions are discussed. The results are compared with the Monte Carlo simulation data. The theory predicts the most important features of the retention process. PMID- 19296625 TI - Essential state models for solvatochromism in donor-acceptor molecules: the role of the bridge. AB - Essential state models are presented to discuss absorption spectra of two related donor-acceptor (DA) chromophores that show two solvatochromic bands in the near infrared spectral region. The two-state model only accounts for the lowest energy band and results in a very small value of mu(0), the dipole moment associated with the D(+)A(-) state. The model is then extended to account for the active role of the bridge: the resulting three-state model satisfactorily reproduces the double solvatochromism, leading at the same time to a roughly doubled estimate of mu(0). This result, supported by a detailed analysis of an N-state model that explicitly accounts for bridge states, rationalizes the well-known discrepancy between the geometrical DA distance and the dipole length extracted from the analysis of optical spectra of DA chromophores as reflecting the active role of bridge states, not explicitly accounted for in essential state models. PMID- 19296626 TI - Ultrafast excited-state dynamics of kynurenine, a UV filter of the human eye. AB - The excited-state dynamics of kynurenine (KN) has been examined in various solvents by femtosecond-resolved optical spectroscopy. The lifetime of the S(1) state of KN amounts to 30 ps in aqueous solutions, increases by more than 1 order of magnitude in alcohols, and exceeds 1 ns in aprotic solvents such as DMSO and DMF, internal conversion (IC) being shown to be the main deactivation channel. The IC rate constant is pH independent but increases with temperature with an activation energy of about 7 kJ/mol in all solvents studied. The dependence on the solvent proticity together with the observation of a substantial isotope effect indicates that hydrogen bonds are involved in the rapid nonradiative deactivation of KN in water. These results give new insight into the efficiency of KN as a UV filter and its role in cataractogenesis. PMID- 19296627 TI - Theoretical study of dynamics for the abstraction reaction H' + HBr(v=0, j=0) --> H'H + Br. AB - Theoretical studies of the dynamics of the abstraction reaction, H' + HBr (v=0,j=0) --> H'H + Br, have been performed with quasiclassical trajectory method (QCT) on a new ab initio potential energy surface (Y. Kurosaki and T. Takayanagi, private communication). The calculated QCT cross sections are in good agreement with earlier quantum wave packet results over most of the collision energy range from 0.1 to 2.6 eV, and the state-resolved rotational distributions of the product H'H molecule are quantitatively consistent with the experimental results. Comparisons of the QCT-calculated rotational-state-resolved cross sections on different potential energy surfaces show that the characteristics of the potential energy surface in the region far away from the minimum energy path have a large influence on the title abstraction reaction dynamics, and the indirect reactions that do not follow the minimum energy path have little influence on the differential cross sections (DCS). The DCSs are mainly governed by the direct reactions that do follow the minimum energy path, at both low and high collision energies. The degree of the rotational alignment of the product H'H molecule is strong at high collision energies, which means that the influence of the indirect reactions on the product rotational alignment is negligible, whereas the distribution of P(varphi(r)) is sensitive to the indirect reactions at high collision energies. With increasing collision energy, the polarization of the product rotational angular momentum decreases and the molecular rotation of the product prefers an in-plane reaction mechanism rather than the out-of-plane mechanism. PMID- 19296628 TI - Oxidative addition of ammonia at a silicon(II) center and an unprecedented hydrogenation reaction of compounds with low-valent group 14 elements using ammonia borane. AB - The reaction of L'Si [L' = CH{(C=CH(2))(CMe)(2,6-iPr(2)C(6)H(3)N)(2)}] with ammonia under oxidative addition results in L'Si(H)NH(2). H(3)NBH(3) functions as a source of hydrogen for the hydrogenation of N-heterocyclic carbenes and germylenes. PMID- 19296629 TI - Phases of solid methanol. AB - The solid phases of methanol were investigated using IR spectroscopy and numerical calculations with the SIESTA method. Improved spectra are reported of amorphous methanol at 90 K, and in particular of the alpha and beta phases at 130 and 165 K, respectively, with assignments of bands not previously measured. The main features of the spectra of each phase are discussed and compared. A study of spectral changes with temperature leads to the conclusion that the metastable phase previously reported might be a mixture of the two known stable phases. Such a mixture could explain all spectral features observed in this investigation. The theoretical calculations provide reasonable agreement with the experimental data for most of the parameters, but predict H-bondings stronger than those observed. Differences between the spectra of the alpha and beta phases are predicted with similar characteristics to the experimental results. PMID- 19296630 TI - Accurate double many-body expansion potential energy surface for the lowest singlet state of methylene. AB - A single-sheeted double many-body expansion potential energy surface is reported for ground-state CH(2) by fitting accurate ab initio energies that have been semiempirically corrected by the double many-body-expansion scaled-external correlation method. The energies of about 2500 geometries have been calculated using the multireference configuration interaction method, with the single diffusely corrected aug-cc-pVQZ basis set of Dunning and the full valence complete active space wave function as reference. The topographical features of the novel global potential energy surface are examined and compared with other potential energy surfaces. PMID- 19296631 TI - Interlocked host anion recognition by an indolocarbazole-containing [2]rotaxane. AB - The design, synthesis, structure, and anion-binding properties of the first indolocarbazole-containing interlocked structure are described. The novel [2]rotaxane molecular structure incorporates a neutral indolocarbazole-containing axle component which is encircled by a tetracationic macrocycle functionalized with an isophthalamide anion recognition motif. (1)H NMR and UV-visible spectroscopies and X-ray crystallography demonstrated the importance of pi-donor acceptor, CH...pi, and electrostatic interactions in the assembly of pseudorotaxanes between the electron-deficient tetracationic macrocycle and a series of pi-electron-rich indolocarbazole derivatives. Subsequent urethane stoppering of one of these complexes afforded a [2]rotaxane, which was shown by (1)H NMR spectroscopic titration experiments to exhibit enhanced chloride and bromide anion recognition compared to its non-interlocked components. Computational molecular dynamics simulations provide further insight into the mechanism and structural nature of the anion recognition process, confirming it to involve cooperative hydrogen-bond donation from both macrocycle and indolocarbazole components of the rotaxane. The observed selectivity of the [2]rotaxane for chloride is interpreted in terms of its unique interlocked binding cavity, defined by the macrocycle isophthalamide and indolocarbazole N-H protons, which is complementary in size and shape to this halide guest. PMID- 19296632 TI - Direct-dynamics study of the F + CH4, C2H6, C3H8, and i-C4H10 reactions. AB - We present a theoretical study of the dynamics of the first few members of the F + alkane --> HF + alkyl family of reactions (alkane = CH(4), C(2)H(6), C(3)H(8), and i-C(4)H(10)). Quasiclassical trajectories have been propagated employing a reparameterized semiempirical Hamiltonian that was derived in this work based on ab initio information of the global potential-energy surfaces of all reactions studied. The accuracy of the Hamiltonian is probed via comparison of the calculated dynamics properties with experimental results in the F + CH(4) --> HF + CH(3), F + CD(4) --> DF + CD(3), and F + C(2)H(6) --> HF + C(2)H(5) reactions. Additional calculations on the F + C(3)H(8) --> HF + C(3)H(7) and F + i-C(4)H(10) --> HF + C(4)H(9) reactions have been analyzed with emphasis on the difference in the dynamics of reactions occurring at primary, secondary, and tertiary sites. We learn that at low collision energies, the amount of energy going into HF vibration increases very slightly along the primary --> secondary --> tertiary sequence. In addition, reactions involving larger alkane molecules tend to channel more energy toward alkyl products at the expense of the rest of the degrees of freedom. Angular distributions are also dependent on the abstraction site, with tertiary abstractions resulting in slightly more backward scattering than reactions at primary sites. PMID- 19296633 TI - Structures and properties of closed ladderanes C24H24, laddersilanes Si24H24, and their nitrogen-containing isoelectronic equivalents: a G3(MP2) investigation. AB - An investigation has been undertaken to study the large closed ladderanes C(24)H(24) and their analogs. Thirteen isoelectronic species have been identified as local minima on the MP2(FU)/6-31G(d) potential energy surface, including three C(24)H(24), four Si(24)H(24), two N(24,) and four C(24-x)H(24-x)N(x) (x = 6, 8, 12) isomers. Of these 13 species, 11 are reported for the first time. Their structures, stabilities, HOMO-LUMO gaps, and G3(MP2) heats of formations are computationally obtained and discussed. These results are also compared with those of the related species already available in the literature. Our results show that molecules with 12-membered rings are highly energetic and much less stable than their corresponding isomers that do not have such rings. Isomer II of Si(24)H(24) with anticonformations has a very small HOMO-LUMO gap of 2.56 eV, approaching the semiconductor range. Therefore, it is a candidate of potential semiconductor materials. Meanwhile, the N(24) and other nitrogen-containing species are candidates for high-energy density materials. Our results also indicate that C(18)H(18)N(6) and C(16)H(16)N(8) may be good hexa- and octadentate ligands for metal cations. PMID- 19296634 TI - Simplest neutral singlet C2E4 (E = Al, Ga, In, and Tl) global minima with double planar tetracoordinate carbons: equivalence of C2 moieties in C2E4 to carbon centers in CAl4(2-) and CAl5(+). AB - Ab initio and DFT calculations have been carried out to search for the simplest neutral singlet species with double planar tetracoordinate carbons (dptCs) [the "simplest" means the species containing the least number (six) and types (two) of atoms]. Under the restrictions to the possible models (M1-M4) with dptCs and to the singlet electronic states, the B3LYP/6-31+G* scanning on the candidates, C(2)E(4) (E = the second- and third-row main group elements), only led to two minima (D(2h) C(2)Al(4) and C(2h) C(2)Be(4)) with stable DFT wave functions. The extensions to the heavier elements after the fourth row in the IIA and IIIA groups revealed that the D(2h) C(2)E(4) (E = Ga, In, and Tl) are also minima with dptCs but C(2)Ca(4) (C(2h)) is a first-order saddle point. Extensive explorations at the DFT level on their potential energy surfaces (PESs) further confirmed that the D(2h) C(2)E(4) (E = Al, Ga, In, and Tl) are the global minima, but the C(2h) C(2)Be(4) is a local minimum. The optimizations at the MP2 level distorted the D(2h) C(2)E(4) (E = Ga, In, and Tl) slightly and the distortion energies are less than 0.02 kcal/mol. The C(2)E(4) (E = Al, Ga, In, and Tl) with dptCs are 18.0, 18.3, 13.4, and 12.2 kcal/mol energetically more favorable than their nearest isomers, respectively, at the CCSD(T)//MP2 level with aug-cc-pVTZ for C and Al and aug-cc-pVTZ-PP for Ga, In, and Tl basis set. The substantial energy differences suggest their promise to be experimentally realized. The strong peak on the C(2)Al(4)(-) component in the time-of-flight mass spectrum from laser vaporization of a mixed graphite/aluminum may relate to the D(2h) C(2)Al(4) global minimum. The analyses of the electronic structures of C(2)Al(4) (D(2h)), CAl(4)(2-) (D(4h)) and CAl(5)(+)(D(5h)) indicates that the C(2) moiety in C(2)Al(4) is the equivalence of carbon centers in CAl(4)(2-) and CAl(5)(+) and unveils the reasons for their stability. The electronic structures of C(2)Al(4) and ethene are compared. On the one hand, an Al atom functions like an H atom because the eight more valence electrons of C(2)Al(4) than C(2)H(4) occupy four nonbonding orbitals and are not effectively utilized for bonding. On the other hand, an Al atom is different from an H atom because an Al atom has p electrons available for peripheral bonding around the C(2) moieties in C(2)Al(4), which further rationalize the origins for C(2)E(4) to achieve double ptCs. PMID- 19296635 TI - Electronic structure and TDDFT optical absorption spectra of silver nanorods. AB - Density functional theory calculations are employed to determine optimized geometries and excitation spectra for small pentagonal silver nanorods Ag(n), with n = 13, 19, 25, 31, 37, 43, 49, 55, 61, and 67 in various charge states. The asymptotically correct SAOP functional is utilized in the excitation calculations. Silver nanorods exhibit a sharp longitudinal excitation that results from a mixture of orbital transitions; the wavelength for this excitation depends linearly on the length of the nanorod. The broad transverse excitation arises from multiple excited states. A particle-in-a-box model is employed to explain the linear dependence of the longitudinal excitation wavelength on nanorod length. PMID- 19296636 TI - Chemoselective C-H bond activation: ligand and solvent free iron-catalyzed oxidative C-C cross-coupling of tertiary amines with terminal alkynes. Reaction scope and mechanism. AB - FeCl(2) catalyzes the oxidative C-C cross-coupling of tertiary amines with terminal alkynes into propargylamines using (t-BuO)(2) as oxidant. The reaction can be applied to aromatic and aliphatic amines and alkynes without solvent. High chemoselectivity for aminomethyl groups is due to a steric factor. PMID- 19296637 TI - Structure and aromaticity of B6H5(+) cation: a novel borhydride system containing planar pentacoordinated boron. AB - A novel structural pattern: two-dimensional, five-membered ring-like boron hydride system B(6)H(5)(+), which contains both planar tetra- and pentacoordinated boron (ptB and ppB) was designed and investigated at [B3LYP, MP2(Full) and CCSD(T,FC)]/6-311+G(d,p) theoretical levels. The results indicate that both degenerate ppB B(6)H(5)(+) configurations A (D(5H),(1)A(1)') and B (C(2nu),(1)A(1)) are energetically favorable, sharing the lowest energy comparison with other isomers considered in this work. The computed considerably large smallest vibrational frequencies [197.5 and 197.4 cm(-1), respectively at MP2 (full) level], HOMO-LUMO energy separations [10.99-11.03 eV computed at MP2 (full) level], and fairly large endothermic dissociation reaction energies suggest they are promising candidate molecules for experimental detection. The detailed molecular orbital analysis, natural bond orbital analysis, magnetic susceptibility anisotropy, and NMR chemical shift analyses confirmed that the two low-energy ppB molecules exhibit strong aromaticities. PMID- 19296638 TI - Nagelamides Q and R, novel dimeric bromopyrrole alkaloids from sponges Agelas sp. AB - Two new dimeric bromopyrrole alkaloids, nagelamides Q (1) and R (2), have been isolated from Okinawan marine sponges of the genus Agelas (SS-1134 and SS-956, respectively), and the structures and stereochemistry were elucidated from spectroscopic data. Nagelamide Q (1) is a rare dimeric bromopyrrole alkaloid possessing a pyrrolidine ring, while nagelamide R (2) is the first bromopyrrole alkaloid having an oxazoline ring. Nagelamides Q (1) and R (2) showed antimicrobial activity. PMID- 19296639 TI - Nickel-catalyzed biaryl coupling of heteroarenes and aryl halides/triflates. AB - Ni-based catalytic systems for the arylation of heteroarenes with aryl halides and triflates have been established. Ni(OAc)(2)/bipy is a general catalyst for aryl bromides/iodides, and Ni(OAc)(2)/dppf is effective for aryl chlorides/triflates. Thiazole, benzothiazole, oxazole, benzoxazole, and benzimidazole are applicable as heteroarene coupling partners. A rapid synthesis of febuxostat, a drug for gout and hyperuricemia, is also demonstrated. PMID- 19296640 TI - Useful modified cellulose polymers as new emulsifiers of cubosomes. AB - This report introduces modified cellulose polymers as new emulsifiers of cubosomes. We prepared novel nanoparticles containing cubic-phase-forming lipids using hydroxypropyl methylcellulose acetate succinate (HPMCAS). Small-angle X-ray scattering showed a much lower incorporation of HPMCAS into the cubic structure of monoolein than did a conventional emulsifier, Pluronic F127, which is known to modify the cubic structure. Cubosomes prepared with HPMCAS showed roughly equal stability as nanoparticles with Pluronic F127. These results suggest that HPMCAS can be a novel emulsifier of cubosomes, which brings about no internal structure modification. PMID- 19296641 TI - Investigation of catalytic characterization of two-dimensional molecular space with regular ammonium and pyridine groups. AB - Novel two-dimensional molecular space with regular pyridine groups layered pyridine-4-amidepropylsilica (PAPS) and pyridine-4-amidephenylsilica (PAPhS) were successfully synthesized through grafting pyridine groups in the layer structure of two-dimensional molecular space with regular ammonium groups layered aminopropylsilica (ATMS-DS) and layered aminophenylsilica (APhTMS-DS). The two dimensional structures were kept after grafting reaction of pyridine groups in PAPS and PAPhS. The catalytic potentials of two-dimensional molecular space with regular ammonium and pyridine groups were investigated. The catalytic capability of APhTMS-DS, PAPS, and PAPhS was confirmed through Knoevenagel condensation reactions. Knoevenagel condensation of aromatic aldehydes with malononitrile was not observed in the presence of ATMS-DS. Otherwise, the lower yield of Knoevenagel condensation of higher active 2-chlorobenzaldehyde with malononitrile in the presence of APhTMS-DS, PAPS, and PAPhS indicated the potential of the two dimensional molecular space with regular catalyst molecules on influencing catalysis processes utilizing the chemical and geometrical limits. PMID- 19296642 TI - Palladium(II)-catalyzed asymmetric coupling of allylic alcohols and vinyl ethers: insight into the palladium and copper bimetallic catalyst. AB - The use of PdX(2), CuX(2) (X = OAc, OCOCF(3), or Cl), and catechol together with (S,S)-4,4'-bisbenzyl-2,2'-bioxazoline under O(2) induces the asymmetric coupling of cinnamyl alcohols and vinyl ethers to give (R)-(+)-2-alkoxy-4 benzylidenetetrahydrofurans in 40-53% ee (79% yield). The present study provides implications for the so-called Wacker catalyst of PdX(2) and CuX(2), in terms of that the anionic ligands (X) of Pd and Cu interchange between two metals. PMID- 19296643 TI - The orientation of BN-related peptides adsorbed on SERS-active silver nanoparticles: comparison with a silver electrode surface. AB - We used surface-enhanced Raman scattering (SERS) to characterize the adsorption behavior of bombesin (BN) and five BN-related peptides, including phyllolitorin, [Leu(8)]phyllolitorin, neuromedin C (NMC), neuromedin B (NMB), and PG-L (Pseudophryne guntheri), in a silver colloidal solution. Our experiments show that the pyrrole coring of the Trp and aromatic ring of Phe of these peptides are preferentially adsorbed on silver nanoparticles. However, the geometry of the rings and the strength of the interactions with this surface vary among peptides. Additionally, these peptides are weakly coordinated to the colloidal silver surface through the CO fragment of a peptide bond, between Gln/Leu/His and Trp residues, and CNC and SC fragments. Also, using the recently reported SERS spectra of these peptides immobilized onto an electrochemically roughened silver electrode surface, we demonstrate substrate-induced changes in the adsorption behavior of these peptides. Comparative analysis indicates that the interactions between peptides and the enhancing surfaces depend strongly on the geometry of the Trp, CONH, and SC fragments of these biomolecules etched on the surfaces. PMID- 19296644 TI - Connectivity and proximity between quadrupolar nuclides in oxide glasses: insights from through-bond and through-space correlations in solid-state NMR. AB - The connectivity and proximity among framework cations and anions in covalent oxide glasses yields unique information whereby their various transport and thermodynamic properties can be predicted. Recent developments and advances in the reconstruction of anisotropic spin interactions among quadrupolar nuclides (spin > (1)/(2)) in solid-state NMR shed light on a new opportunity to explore local connectivity and proximity in amorphous solids. Here, we report the 2D through-bond (J-coupling) and through-space (dipolar coupling) correlation NMR spectra for oxide glasses where previously unknown structural details about the connectivity and proximity among quadrupolar nuclides ((27)Al, (17)O) are determined. Nonbridging oxygen peaks in Ca-aluminosilicate glasses with distinct connectivity, such as Ca-O-Al and Al-O-(Al, Si) are well distinguished in {(17)O}(27)Al solid HMQC NMR spectra. Both peaks shift to a lower frequency in direct and indirect dimensions upon the addition of Si to the Ca-aluminate glasses. The 2D (27)Al double quantum magic angle spinning NMR spectra for Mg aluminoborate glasses indicate the preferential proximity between ([4])Al and ([5])Al leading to the formation of correlations peaks such as ([4])Al-([4])Al, ([4])Al-([5])Al, and ([5])Al-O-([5])Al. A fraction of the ([6])Al-([6])Al correlation peak is also noticeable while that of ([4,5])Al-([6])Al is missing. These results suggest that ([6])Al is likely to be isolated from the ([4])Al and ([5])Al species, forming ([6])Al clusters. The experimental realization of through-bond and through-space correlations among quadrupolar nuclides in amorphous materials suggests a significant deviation from the random distribution among framework cations and a spatial heterogeneity due to possible clustering of framework cations in the model oxide glasses. PMID- 19296645 TI - Experimental and theoretical study of hydrogen thiocarbonate for heterogeneous reaction of carbonyl sulfide on magnesium oxide. AB - In situ diffuse reflectance infrared Fourier transform spectroscopy combined with derivative spectroscopy analysis, two-dimensional correlation spectroscopy analysis, and quantum chemical calculations were used to investigate the infrared absorbance assignment and the molecular structure of hydrogen thiocarbonate on magnesium oxide. The bands at 1283 and 1257 cm(-1), which had the typical characteristic of intermediate, were observed in experiments for the heterogeneous reaction of COS on MgO. On the basis of two-dimensional correlation spectroscopy analysis and quantum chemical calculations, the band at 1283 cm(-1) was assigned to the v(s) band of bridged thiocarbonate which formed on the two neighboring Mg atoms in the (100) face of MgO crystal, and the band at 1257 cm( 1) was the v(s) band of monodentate thiocarbonate on MgO. The v(as)(OCO) band of thiocarbonates was invisible in the experiment due to their weak absorbance and the interruption of surface carbonate. The formation mechanism of thiocarbonates is proposed, which occurred through a nucleophilic attack of preadsorbed COS by surface -OH groups followed by hydrogen atom transfer from the -OH group to the sulfur atom of preadsorbed COS. The activation energy for the intramolecular proton-transfer reaction of bridged thiocarbonate was calculated to be 18.52 kcal x mol(-1) at the B3LYP/6-31+G(d,p) level of theory. PMID- 19296646 TI - Haliclonin A, a new macrocyclic diamide from the sponge Haliclona sp. AB - Haliclonin A (1), a macrocyclic diamide of a novel skeletal class, was isolated from the marine sponge Haliclona sp. collected from Korean waters. The structure of this compound was determined using a combination of spectroscopic and chemical analyses. The new compound exhibited moderate cytotoxicity and antibacterial activity against diverse microbial strains. PMID- 19296647 TI - Plasmonic electromagnetic hot spots temporally addressed by photoinduced molecular displacement. AB - We report the observation of temporally varying electromagnetic hot spots in plasmonic nanostructures. Changes in the field amplitude, position, and spatial features are induced by embedding plasmonic silver nanorods in the photoresponsive azo-polymer. This polymer undergoes cis-trans isomerization and wormlike transport within resonant optical fields, producing a time-varying local dielectric environment that alters the locations where electromagnetic hot spots are produced. Finite-difference time-domain and Monte Carlo simulations that model the induced field and corresponding material response are presented to aid in the interpretation of the experimental results. Evidence for propagating plasmons induced at the ends of the rods is also presented. PMID- 19296648 TI - Straightforward and highly efficient catalyst-free one-step synthesis of 2-(purin 6-yl)acetoacetic acid ethyl esters, (purin-6-yl)acetates, and 6-methylpurines through S(N)Ar-based reactions of 6-halopurines with ethyl acetoacetate. AB - A novel approach to the synthesis of purines bearing functionalized carbon substituents or methyl in position 6 was developed. Under different reaction conditions, 6-halopurine derivatives could react with ethyl acetoacetate efficiently to yield 2-(purin-6-yl)acetoacetic acid ethyl esters, (purin-6 yl)acetates and 6-methylpurines respectively. No metal catalyst and ligand were required. PMID- 19296649 TI - Preparation of diphenyl phosphide and substituted phosphines using alkali metal in silica gel (M-SG). AB - Alkali metals absorbed in silica gel (the M-SG reagents) efficiently cleave C-P bonds in triaryl- and diarylphosphines. The resulting alkali metal phosphides can serve as useful building blocks for a variety of phosphines. Alkyldiarylphosphines undergo exclusive aryl group cleavage. PMID- 19296650 TI - Binding site detection and druggability index from first principles. AB - In drug discovery, it is essential to identify binding sites on protein surfaces that drug-like molecules could exploit to exert a biological effect. Both X-ray crystallography and NMR experiments have demonstrated that organic solvents bind precisely at these locations. We show that this effect is reproduced using molecular dynamics with a binary solvent. Furthermore, analysis of the simulations give direct access to interaction free energies between the protein and small organic molecules, which can be used to detect binding sites and to predict the maximal affinity that a drug-like molecule could attain for them. On a set of pharmacologically relevant proteins, we obtain good predictions for druggable sites as well as for protein-protein and low affinity binding sites. This is the first druggability index not based on surface descriptors and, being independent of a training set, is particularly indicated to study unconventional targets such as protein-protein interactions or allosteric binding sites. PMID- 19296651 TI - Identification of a metabolically stable triazolopyrimidine-based dihydroorotate dehydrogenase inhibitor with antimalarial activity in mice. AB - Plasmodium falciparum causes 1-2 million deaths annually. Yet current drug therapies are compromised by resistance. We previously described potent and selective triazolopyrimidine-based inhibitors of P. falciparum dihydroorotate dehydrogenase (PfDHODH) that inhibited parasite growth in vitro; however, they showed no activity in vivo. Here we show that lack of efficacy against P. berghei in mice resulted from a combination of poor plasma exposure and reduced potency against P. berghei DHODH. For compounds containing naphthyl (DSM1) or anthracenyl (DSM2), plasma exposure was reduced upon repeated dosing. Phenyl-substituted triazolopyrimidines were synthesized leading to identification of analogs with low predicted metabolism in human liver microsomes and which showed prolonged exposure in mice. Compound 21 (DSM74), containing p-trifluoromethylphenyl, suppressed growth of P. berghei in mice after oral administration. This study provides the first proof of concept that DHODH inhibitors can suppress Plasmodium growth in vivo, validating DHODH as a new target for antimalarial chemotherapy. PMID- 19296652 TI - Inhibition of heat shock induction of heat shock protein 70 and enhancement of heat shock protein 27 phosphorylation by quercetin derivatives. AB - Inhibitors of heat-induced heat shock protein 70 (HSP70) expression have the potential to enhance the therapeutic effectiveness of heat-induced radiosensitization of tumors. Among known small molecule inhibitors, quercetin has the advantage of being easily modified for structure-activity studies. Herein, we report the ability of five monomethyl and five carbomethoxymethyl derivatives of quercetin to inhibit heat-induced HSP70 expression and enhance HSP27 phosphorylation in human cells. While quercetin and several derivatives inhibit HSP70 induction and enhance HSP27 phosphorylation at Ser78, other analogues selectively inhibit HSP70 induction without enhancing HSP27 phosphorylation that would otherwise aid in cell survival. We also show that good inhibitors of HSP70 induction are also good inhibitors of both CK2 and CamKII, kinases that are known to activate HSP70 expression by phosphorylation of heat shock transcription factor 1. Derivatives that show poor inhibition of either or both kinases are not good inhibitors of HSP70 induction, suggesting that quercetin's effectiveness is due to its ability to inhibit both kinases. PMID- 19296653 TI - Discovery of N-(4-methoxyphenyl)-N,2-dimethylquinazolin-4-amine, a potent apoptosis inducer and efficacious anticancer agent with high blood brain barrier penetration. AB - As a continuation of our structure-activity relationship (SAR) studies on 4 anilinoquinazolines as potent apoptosis inducers and to identify anticancer development candidates, we explored the replacement of the 2-Cl group in our lead compound 2-chloro-N-(4-methoxyphenyl)-N-methylquinazolin-4-amine (6b, EP128265, MPI-0441138) by other functional groups. This SAR study and lead optimization resulted in the identification of N-(4-methoxyphenyl)-N,2-dimethylquinazolin-4 amine (6h, EP128495, MPC-6827) as an anticancer clinical candidate. Compound 6h was found to be a potent apoptosis inducer with EC(50) of 2 nM in our cell-based apoptosis induction assay. It also has excellent blood brain barrier penetration, and is highly efficacious in human MX-1 breast and other mouse xenograft cancer models. PMID- 19296654 TI - Water-stable diblock polystyrene-block-poly(2-vinyl pyridine) and diblock polystyrene-block-poly(methyl methacrylate) cylindrical patterned surfaces inhibit settlement of zoospores of the green alga Ulva. AB - Nanopatterned surfaces with hydrophobic and hydrophilic domains were produced using the diblock copolymer polystyrene-block-poly(2-vinyl pyridine) (PS-b-P2VP) and polystyrene-block-poly(methyl methacrylate) (PS-b-PMMA). The PS-b-P2VP diblock copolymer, mixed with the cross-linker benzophenone and spin-coated onto silicon wafers, showed self-assembled cylindrical structures, which were retained after UV treatment for cross-linking. The thin films displayed cylindrical domains after immersion in water. This study shows that pattern retention in water is possible for a long period of time, at least for two weeks in pure water and three weeks in artificial seawater. The PS-b-PMMA diblock showed self assembled cylindrical structures. PS-b-P2VP and PS-b-PMMA cylindrical patterned surfaces showed reduced settlement of zoospores of the green alga Ulva compared to unpatterned surfaces. The copolymers were investigated using atomic force microscopy and X-ray photoelectron spectroscopy. PMID- 19296655 TI - The Drosophila transcription factor ultrabithorax self-assembles into protein based biomaterials with multiple morphologies. AB - The use of proteins as monomers for materials assembly enables customization of chemical, physical, and functional properties. However, natural materials-forming proteins are difficult to produce as recombinant protein monomers and require harsh conditions to initiate assembly. We have generated materials using the recombinant transcription factor Ultrabithorax, a Drosophila melanogaster protein not known or anticipated to form extended oligomers in vivo. Ultrabithorax self assembles at the air-water interface into nanoscale fibers, which further associate to form macroscale films, sheets, ropes, and tethered encapsulates. These materials self-adhere, allowing construction of more complex architectures. The Ultrabithorax sequence contains two regions capable of generating materials, only one of which contains motifs found in elastomeric proteins. However, both minimal regions must be included to produce robust materials. Relative to other protein-based materials, Ultrabithorax assembles at significantly reduced concentrations, on faster timescales, and under gentler conditions, properties that facilitate future materials engineering and functionalization. PMID- 19296656 TI - Injectable poly(amidoamine)-poly(ethylene glycol)-poly(amidoamine) triblock copolymer hydrogel with dual sensitivities: pH and temperature. AB - A novel triblock copolymer for use in an injectable pH- and temperature-sensitive hydrogel is synthesized by conjugating poly(amidoamine) (PAA) to poly(ethylene glycol): poly(amidoamine)-poly(ethylene glycol)-poly(amidoamine) (PAA-PEG-PAA). The polymer was characterized with (1)H NMR and gel permeation chromatography in the diluents CDCl(3) and CHCl(3), respectively. The PAA block acts as a pH- and temperature-sensitive block. The PAA-PEG-PAA copolymer in aqueous solution (12.5 wt %) underwent a sol-gel transition as a function of pH and temperature. After injection into a rat, the copolymer solution (12.5 wt %) was immediately changed to a gel. PMID- 19296657 TI - Effect of impeller speed and pH on the production of poly(3-hydroxybutyrate) using Bacillus cereus SPV. AB - P(3HB), is one of the most well studied polyhydroxyalkanoates. It is biodegradable, biocompatible, exhibits thermoplastic properties and can be produced from renewable carbon sources. The commercial exploitation of P(3HB) has been mainly held back by its high production costs. Hence, a lot of research is required to optimize P(3HB) fermentation conditions. In this study we have focused on the effects of impeller speed and pH on P(3HB) production in Bacillus cereus SPV. Four different impeller speeds, 50, 125, 250, and 500 rpm, were used. The highest amount of P(3HB) accumulation was achieved using 125 rpm impeller speed (34% dcw) and this was attributed to optimal cell growth rate. Also, pH stat fermentations were carried out at pH 3.0, 6.8, and 10. This study confirmed that lack of P(3HB) degradation during unbuffered Bacillus fermentations is due to the low pH conditions. This observation is crucial for the industrial exploitation of the genus Bacillus for the production of P(3HB). PMID- 19296658 TI - Peptide-poly(L-lysine citramide) conjugates and their in vitro anti-HIV behavior. AB - Poly(L-lysine citramide) is a degradable bioresorbable polyanion whose polyamide chains are composed of citric acid and L-lysine building blocks. Its chemical and physicochemical properties were extensively investigated in the past for its interest as polymeric drug carrier. In this work, 4(S)-amino-3-(S)-hydroxy-5 phenylpentanoyl-isoleucyl-phenylalanine methyl ester, a pseudopeptide active against the HIV protease in vitro, was linked to poly(L-lysine citramide) in attempts to promote solubility and cell penetration. Conjugates were characterized by FTIR, NMR, SEC, DLS, amino-acid analyses, and toxicity in mice. They degraded slowly at pH 7.4 and more rapidly at pH 4.5, two pH values selected to mimic extra-cellular fluids and intralysosome medium, respectively. According to capillary zone electrophoresis, degradation did not release the peptide. The phenylalanyl-isoleucyl-phenylalanine methyl ester peptide, inactive against the protease in vitro, was used as negative control. The anti-HIV activities of the carrier, of the conjugates and of model molecules, including a fluorescence labeled pseudopeptide conjugate, were evaluated comparatively in vitro using two cell lines, namely, CEM-SS and MT-4 cells, infected with HIV-1 LAI and IIIB isolates, respectively. Unexpectedly, all the conjugates showed in vitro antiviral activity independent of peptide release and of inhibition of the HIV protease. According to FACS analysis, the antiviral activity was related to the presence of peptide moieties along the polymer chains and depended on the order by which cells, viruses, and conjugates were presented to each other. Although it was not possible to determine whether the antiviral activity resulted from interactions between conjugates and cells or conjugates and virus or both, the conjugates appeared able to inhibit the binding of the virus to cells in vitro when introduced before cell infection. None of the conjugates exhibited acute toxicity in mice. PMID- 19296659 TI - Ru(II) catalysts supported by hydridotris(pyrazolyl)borate for the hydroarylation of olefins: reaction scope, mechanistic studies, and guides for the development of improved catalysts. AB - Carbon-carbon bond formation is the central method by which synthetic chemists add complexity, which often represents value, to molecules. Uniting a carbon chain with an aromatic substrate to yield an alkyl arene product is thus a molecular means of creating value-added materials. A traditional method for generating alkyl arenes is Friedel-Crafts catalysis, in which an alkyl halide or olefin is activated to react with an aromatic substrate. Unfortunately, despite the development of new generations of solid-state catalysts, the reaction often requires relatively harsh conditions and frequently gives poor to moderate selectivity. Conversely, a halide can first be incorporated into the aromatic ring, and the aryl halide can subsequently be joined by a variety of catalytic coupling techniques. But generating the aryl halide itself can be problematic, and such methods typically are not atom-economical. The addition of aromatic C-H bonds across the C-C double bonds of olefins (olefin hydroarylation) is therefore an attractive alternative in the preparation of alkyl arenes. Despite the dominance and practical advantages of heterogeneous catalysts in industrial synthesis, homogeneous systems can offer an enhanced ability to fine-tune catalyst activity. As such, well-defined homogeneous catalysts for the hydroarylation of olefins provide a potentially promising avenue to address issues of selectivity, including the production of monoalkylated arene products and the control of linear-to-branched ratios for synthesis of long-chain alkyl arenes, and provide access to more ambient reaction conditions. However, examples of homogeneous catalysts that are active for the conversion of unactivated aromatic and olefin substrates to alkyl arene products that function via metal mediated C-H activation pathways are limited. In this Account, we present results from research aimed at the development of Ru(II) catalysts supported by the hydridotris(pyrazolyl)borate (Tp) ligand for the addition of aromatic C-H bonds across olefins. On the basis of detailed mechanistic studies with TpRu(L)(NCMe)R catalysts, in which the neutral ancillary ligand L is varied, we have arrived at guidelines for the development of improved catalysts that are based on the octahedral-d6 motif. PMID- 19296660 TI - Comparison of flavonoids and isoflavonoids as antioxidants. AB - The isoflavonoid genistein was found to be a better antioxidant than the isomeric flavonoid apigenin in phosphatidyl liposomes at pH 7.4. The higher antioxidation activity of genistein compared with apigenin is in agreement with its lower oxidation potential (0.73 vs 0.86 V as determined by cyclic voltammetry in aqueous solution of pH= 7.4), lower dissociation enthalpy (87.03 vs 87.88 kcal mol(-1) as calculated for the more reducing 4'-hydroxyl group), and higher radical scavenging capacity in the TEAC assay. On the basis of quantum mechanical calculations for genistein and apigenin in comparison with the flavonoid naringenin and the isoflavonoids puerarin, daidzein, and equol, a lower dipole moment and a larger deviation for the A-to-B dihedral angle from coplanarity (39.3 degrees for genistein, 18.5 degrees for apigenin) are suggested to be important for the increased antioxidant efficiency at water/lipid interfaces among (iso) flavonoids with an equal number of phenolic groups. PMID- 19296661 TI - Pd(II)-catalyzed olefination of electron-deficient arenes using 2,6 dialkylpyridine ligands. AB - Pd(II)-catalyzed meta-olefination of highly electron-deficient arenes is achieved through the use of a rationally designed mutually repulsive ligand. The combination of directed and nondirected C-H functionalization of arenes provides a versatile route for the synthesis of highly sought after 1,2,4-trisubstituted arenes. PMID- 19296662 TI - Photocyclization of tosylstilbenes as a key reaction in the preparation of an analogue of the antitumor agent CC-1065. AB - A photocyclization of tosylstilbenes in the presence of base is used as a key reaction in the synthesis of the CC-1065 analogue 5. PMID- 19296664 TI - Synthesis of 3-aryl-8-oxo-5,6,7,8-tetrahydroindolizines via a palladium-catalyzed arylation and heteroarylation. AB - A selective palladium-catalyzed arylation and heteroarylation of 8-oxo-5,6,7,8 tetrahydroindolizines has been developed. Mechanistic studies assume an electrophilic substitution pathway for this transformation. This method provides an efficient one-step synthesis of 3-aryl-8-oxo-5,6,7,8-tetrahydroindolizines. PMID- 19296665 TI - Diastereoselective synthesis of chiral diporphyrins via intramolecular meso-meso oxidative coupling. AB - Diastereoselective syntheses of meso-meso linked diporphyrins were achieved via intramolecular chiral induction. The structures and conformations were analyzed by CD, UV, NMR, and computational calculations. PMID- 19296666 TI - Indium-catalyzed C-S cross-coupling of aryl halides with thiols. AB - Indium-catalyzed C-S cross-coupling of aromatic and alkane thiols with aryl halides proceeds smoothly in the presence of In(OTf)(3) (10 mol %), TMEDA (20 mol %), and KOH as a base in DMSO at 135 degrees C. When this protocol was utilized, a variety of thiols could be cross-coupled with aryl halides to afford the corresponding aryl sulfides in good to excellent yields. PMID- 19296667 TI - Palladium-catalyzed cocyclotrimerization of allenes with arynes: selective synthesis of phenanthrenes. AB - Palladium-catalyzed cocyclotrimerization of allenes with arynes has been developed for selectively synthesizing phenanthrenes. In the presence of [(allyl)PdCl](2) and P(o-tol)(3), a variety of allenes, including internal and terminal allenes, underwent the cocyclotrimerization with arynes to afford the corresponding phenanthrenes in moderate to good yields. The results showed the selectivity of the reaction based on allenes. PMID- 19296668 TI - Parallel solid-phase synthesis and high-throughput 1H NMR evaluation of a 96 member 1,2,4-trisubstituted-pyrimidin-6-one-5-carboxylic acid library. AB - A solid-phase organic synthesis method has been developed for the preparation of trisubstituted pyrimidin-6-one carboxylic acids 12, which allows elaboration to a 3-dimensional combinatorial library. Three substituents are introduced by initial Knoevenagel condensation of an aldehyde and malonate ester resin 7 to give resin bound 1. Cyclization of 1 with an N-substituted amidine 10, oxidation, and cleavage afforded pyrimidinone 12. The initial solid-phase reaction sequence was followed by gel-phase (19)FNMR and direct-cleavage (1)H NMR of intermediate resins to determine the optimal conditions. The scope of the method for library production was determined by investigation of a 3 x 4 pilot library of twelve compounds. Cyclocondensation of N-methylamidines and 7 followed by CAN oxidation gave mixtures of the resin bound pyrimidin-6-one 11 and the regioisomeric pyrimidin-4-one 15, which after cleavage from the resin afforded a nearly 1:1 mixture of pyrimidin-6-one and pyrimidin-4-one carboxylic acids 12 and 16, respectively. The regiochemical assignment was confirmed by ROESY1D and gHMBC NMR experiments. A library was prepared using 8 aldehydes, 3 nitriles, and 4 amines to give a full combinatorial set of 96 pyrimidinones 12. Confirmation of structural identity and purity was carried out by LCMS using coupled ELS detection and by high-throughput flow (1)H NMR. PMID- 19296669 TI - Tetranortriterpenoids from the Leaves of Cipadessa cinerascens. AB - Seven known and six new tetranortriterpenoids, cineracipadesins A-F (1-6), were isolated from the leaves of Cipadessa cinerascens. Compound 1 has a mexicanolide type structural skeleton with a rare 9alpha,11alpha-epoxide ring; compound 2 has a methyl angolensate-type structure with 9,11-dihydroxy groups, representing the first example of a precursor of a trijugin-type limonoid; and 3 is the first reported methyl angolensate-type limonoid with a ketone group at ring C. Their structures were determined with extensive spectroscopic analysis. X-ray crystallography confirmed the structure of 1. The ability of compounds 1-7 to inhibit the growth of the P-388 murine leukemia cell line was evaluated. PMID- 19296670 TI - Crystal structure of human fibrinogen. AB - A crystal structure of human fibrinogen has been determined at approximately 3.3 A resolution. The protein was purified from human blood plasma, first by a cold ethanol precipitation procedure and then by stepwise chromatography on DEAE cellulose. A product was obtained that was homogeneous on SDS-polyacrylamide gels. Nonetheless, when individual crystals used for X-ray diffraction were examined by SDS gel electrophoresis after data collection, two species of alpha chain were present, indicating that some proteolysis had occurred during the course of operations. Amino-terminal sequencing on post-X-ray crystals showed mostly intact native alpha- and gamma-chain sequences (the native beta chain is blocked). The overall structure differs from that of a native fibrinogen from chicken blood and those reported for a partially proteolyzed bovine fibrinogen in the nature of twist in the coiled-coil regions, likely due to weak forces imparted by unique crystal packing. As such, the structure adds to the inventory of possible conformations that may occur in solution. Other features include a novel interface with an antiparallel arrangement of beta chains and a unique tangential association of coiled coils from neighboring molecules. The carbohydrate groups attached to beta chains are unusually prominent, the full sweep of 11 sugar residues being positioned. As was the case for native chicken fibrinogen, no resolvable electron density could be associated with alphaC domains. PMID- 19296671 TI - Formal total synthesis of N-methylmaysenine. AB - A novel synthetic approach for the formal total synthesis of N-methylmaysenine (1) has been developed. Key steps involve the Ti-mediated vinylogous Mukaiyama aldol reaction of chiral ketene silyl N,O-acetal with beta-dithiane-substituted aldehyde, an aldol condensation, and a ring-closing metathesis reaction. PMID- 19296672 TI - Ganglioside GQ1b: efficient total synthesis and the expansion to synthetic derivatives to elucidate its biological roles. AB - The convergent total synthesis of ganglioside GQ1b based on the "cassette approach" between the nonreducing end GQ1b-core heptasaccharide and glucosylceramide building blocks was accomplished in high overall yield. The use of a sialylalpha(2-->8)sialylalpha(2-->3)galactose sequence as the key building block enhanced the efficiency of the glycan assembly and led to preparative-scale synthesis readily applicable for large-scale preparation. In addition, a judicious choice of p-methoxybenzyl protecting groups on glucosylceramide provided a solution to the previous synthetic problems, including a decrease in the yield of the deprotection steps, and led to elevation of the total yield. Furthermore, unnatural-type GQ1b derivatives were synthesized systematically in good yields by capitalizing on a similar approach in order to elucidate their biological roles. PMID- 19296673 TI - Quantifying green fluorescent protein diffusion in Escherichia coli by using continuous photobleaching with evanescent illumination. AB - Fluorescence recovery after photobleaching and fluorescence correlation spectroscopy are the primary means for studying translational diffusion in biological systems. Both techniques, however, present numerous obstacles for measuring translational mobility in structures only slightly larger than optical resolution. We report a new method using through-prism total internal reflection fluorescence microscopy with continuous photobleaching to overcome these obstacles. Small structures, such as prokaryotic cells or isolated eukaryotic organelles, containing fluorescent molecules are adhered to a surface. This surface is continuously illuminated by an evanescent wave created by total internal reflection. The characteristic length describing the decay of the evanescent intensity with distance from the surface is smaller than the structures. The fluorescence decay rate resulting from continuous evanescent illumination is monitored as a function of the excitation intensity. The data at higher excitation intensities provide apparent translational diffusion coefficients for the fluorescent molecules within the structures because the decay results from two competing processes (the intrinsic photobleaching propensity and diffusion in the small structures). We present the theoretical basis for the technique and demonstrate its applicability by measuring the diffusion coefficient, 6.3 +/- 1.1 microm(2)/s, of green fluorescent protein in Escherichia coli cells. PMID- 19296674 TI - Secretome-based proteomic profiling of Ras-transformed MDCK cells reveals extracellular modulators of epithelial-mesenchymal transition. AB - Epithelial-mesenchymal transition (EMT) is a highly conserved morphogenetic process by which epithelial cells lose their basic morphological characteristics such as cell-cell contact and gain mesenchymal properties such as increased motility and invasiveness. To gain insights into proteins released from cells that modulate the EMT process, we compared secretome protein expression profiles of MDCK cells and Ras-transformed MDCK cells (21D1) that stably express oncogenic Ras using 2D-DIGE/LC-MS/MS. Differentially expressed secretome proteins were compared with their corresponding gene expression profiles using the Affymetrix GeneChip system. Down-regulated proteins were predominantly involved with cell cell contact and cell-matrix adhesion (e.g., desmocollin 2, clusterin, collagen XVII and transforming growth factor-beta induced protein ig-h3), while up regulated proteins were proteases and factors that promote migration (MMP-1, kallikrein 6, TIMP-1, and S100A4/metastasin). Many of the secretome proteins identified in this study have not been previously identified in the context of EMT and may shed light on the underlying mechanisms associated with this cellular process. PMID- 19296675 TI - Targeted systemic delivery of a therapeutic siRNA with a multifunctional carrier controls tumor proliferation in mice. AB - In this study, novel peptide-targeted delivery systems were developed for systemic and targeted delivery of therapeutic siRNA based on a multifunctional carrier, (1-aminoethyl)iminobis[N-(oleicylcysteinylhistinyl-1 aminoethyl)propionamide] (EHCO), which showed pH-sensitive amphiphilic cell membrane disruption. EHCO formed stable nanoparticles with siRNA. Targeted siRNA delivery systems were readily formed by surface modification of the nanoparticles. PEGylation of the siRNA/EHCO nanoparticles significantly reduced nonspecific cell uptake. The incorporation of a bombesin peptide or RGD peptide via a PEG spacer resulted in receptor-mediated cellular uptake and high gene silencing efficiency in U87 cells. Fluorescence confocal microscopic studies demonstrated that EHCO/siRNA nanoparticles and PEG modified EHCO/siRNA nanoparticles were able to facilitate endosomal escape of the siRNA delivery systems. Systemic administration of a therapeutic anti-HIF-1alpha siRNA with the peptide-targeted delivery systems resulted in significant tumor growth inhibition than a nontargeted delivery system or free siRNA via intravenous injection in nude mice bearing human glioma U87 xenografts. The results indicate a great promise of the multifunctional carrier EHCO for systemic and targeted delivery of therapeutic siRNA to treat human diseases with RNAi. PMID- 19296676 TI - Introducing capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) for the characterization of konjac glucomannan oligosaccharides and their in vitro fermentation behavior. AB - The application of capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) as a tool for the characterization of complex carbohydrate structures was investigated for konjac glucomannan (KGM) oligosaccharide mixtures and the monitoring of their structural changes during 72 h of in vitro fermentation with human gut flora. Different types of KGM oligosaccharide mixtures were produced from a KGM polysaccharide using endo-beta-(1,4)-mannanase and endo-beta-(1,4)-glucanase. Distinction of structures emerging from different enzymatic KGM digests and detection of acetylated oligosaccharides were possible by both CE-LIF and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Using CE-LIF it could be shown that the endo beta-(1,4)-glucanase digest exhibited a large degradability of the DP2, DP3, DP5, and DP6 components during in vitro fermentation, whereas the endo-beta-(1,4) mannanase digest was digested only slightly, thereby highlighting the influence of structural characteristics on the fermentability by human gut flora. PMID- 19296677 TI - Alignment-free prediction of polygalacturonases with pseudofolding topological indices: experimental isolation from Coffea arabica and prediction of a new sequence. AB - Polygalacturonases (PGs) have called the attention of microbiology scientists and biotechnology or pharmaceutical industry because they are protein enzymes relevant to phytopathogens invasion, fruit ripening, and potential antimicrobial drug targets. Numeric Topological Indices (TIs) of protein pseudofolding lattices can be used as input for classification algorithms in Quantitative Structure Activity Relationship (OSAR) studies. However, a comparative study of different OSAR models for PGs has not been reported. In this study, we calculated for the first time two classes of TIs (Spectral moments (pik) and Entropy (thetak) values) for the Markov matrices associated to pseudofolding lattices of 108 PGs and 100 non-PGs heterogeneous proteins. Afterward, we developed different linear classifiers based on Linear Discriminant Analysis (LDA) and four types of nonlinear Artificial Neural Networks (ANN). The pik-LDA model correctly classified 98.8% of PGs and 100% non-PGs used to train the model, as well as 98.1% of all sequences used as external validation series. The rk-LDA model was the more accurate and/or simpler found. In addition, we report for the first time the experimental isolation and successful prediction of a new PG sequence from Coffea arabica. This sequence was deposited in the GenBank by our group with accession number GDQ336394. The present type of models are an interesting alignment-free complement to alignment-based procedures. PMID- 19296678 TI - Crystal structures of constitutive nitric oxide synthases in complex with de novo designed inhibitors. AB - New nitric oxide synthase (NOS) inhibitors were designed de novo with knowledge gathered from the studies on the nNOS-selective dipeptide inhibitors. Each of the new inhibitors consists of three fragments: an aminopyridine ring, a pyrrolidine, and a tail of various length and polarity. The in vitro inhibitory assays indicate good potency and isoform selectivity for some of the compounds. Crystal structures of these inhibitors bound to either wild type or mutant nNOS and eNOS have confirmed design expectations. The aminopyridine ring mimics the guanidinium group of L-arginine and functions as an anchor to place the compound in the NOS active site where it hydrogen bonds to a conserved Glu. The rigidity of the pyrrolidine ring places the pyrrolidine ring nitrogen between the same conserved Glu and the selective residue nNOS Asp597/eNOS Asn368, which results in similar interactions observed with the alpha-amino group of dipeptide inhibitors bound to nNOS. These structures provide additional information to help in the design of inhibitors with greater potency, physicochemical properties, and isoform selectivity. PMID- 19296679 TI - Alpha-fluoro-2,2,3,3-tetramethylcyclopropanecarboxamide, a novel potent anticonvulsant derivative of a cyclic analogue of valproic acid. AB - 2,2,3,3-Tetramethylcyclopropanecarboxylic acid (TMCA, 4) is a cyclic analogue of the antiepileptic drug (AED) valproic acid (VPA) (1). alpha-F, alpha-Cl, alpha Br, and alpha-methyl derivatives of 4 and their amides were synthesized and tested in rodent models for anticonvulsant potency and AED-induced teratogenicity. In the anticonvulsant rat-maximal electroshock (MES) and subcutaneous metrazol (scMet) tests, alpha-Cl-TMCD (17) had ED(50) values of 97 and 27 mg/kg, respectively. alpha-F-TMCD (11) was 120 times more potent than VPA in the rat-scMet test (ED(50) = 6 mg/kg) and had a protective index (PI = TD(50)/ED(50)) of 20. In the 6 Hz psychomotor mouse model 11 had ED(50) values of 57 mg/kg (32 mA) and 59 mg/kg (44 mA). The ED(50) values of 11 in the hippocampal kindled rat model and in the pilocarpine-induced-status rat model were 30 and 23 mg/kg, respectively. Unlike 1, 11 was nonteratogenic in mice. This novel compound has the potential to become a candidate for development as a new potent and safe antiepileptic and CNS drug. PMID- 19296680 TI - Palladium catalyzed tandem ring opening-ring closing reaction of diazabicyclic alkenes: a facile one pot strategy for cyclopentannulation of heterocycles. AB - A novel palladium catalyzed protocol for the synthesis of cyclopentene fused heterocycles from diazabicyclic alkenes and ortho-functionalized aryliodides has been elaborated. The reaction can be tuned toward the formation of either 3,4 disubstituted cyclopentenes or cyclopentene fused heterocycles by careful manipulation of the reaction parameters. A number of cyclopentene fused benzofurans and indole derivatives were prepared in excellent yield by utilizing the developed methodology. PMID- 19296681 TI - Coordination of arsine ligands as a general synthetic approach to rare examples of arsenic-antimony and arsenic-bismuth bonds. AB - Tertiary arsines form stable adducts with trichlorostibine and with cationic antimony and bismuth centers, defining coordination chemistry as a general synthetic approach to the formation of rare As-Sb and As-Bi bonds. A series of compounds containing As-Sb and As-Bi bonds define key milestones in the systematic development of interpnictogen compounds. PMID- 19296682 TI - Covalent protein labeling based on noncatalytic beta-lactamase and a designed FRET substrate. AB - Techniques for labeling proteins with small molecules have attracted the attention of many life scientists. We have developed a novel protein labeling system that combines a genetically modified, noncatalytic beta-lactamase variant and specific mechanism-based fluorescent probes. Rational design of the tag protein and the labeling probes enables highly specific incorporation of the fluorogen. The feasibility of our approach was confirmed by gel electrophoresis, mass spectrometry, fluorescence spectroscopy, and fluorescence microscopic imaging. Labeling techniques that satisfy the dual criteria of specificity and fluorogenicity have rarely been reported. As a consequence, this method could be a broadly useful research tool in the field of life science. PMID- 19296683 TI - Columnar mesophases of the complexes of DNA with low-generation poly(amido amine) dendrimers. AB - The electrostatic complexes of polyanionic DNA with dendrimers have been considered as a class of nonviral vector for gene therapy. The gene transfection efficiency has been believed to be influenced by the structure of the complex. In this study, we have systematically characterized the supramolecular structures of the complexes of DNA duplexes with poly(amido amine) (PAMAM) dendrimers with generation two (G2) and three (G3) in pure water using small-angle X-ray scattering. The structures were examined as a function of the charge density of the dendrimer expressed by its degree of protonation (dp) and the molar ratio of the amine groups of dendrimer to the phosphate groups of DNA (N/P). The DNA chains in all complexes under study were found to self-organize into two dimensional hexagonal or square lattice. In general, hexagonal phase was the favorable structure for G2 complexes, while the DNA in G3 complexes tended to organize into a square lattice. Interesting transitions between the columnar mesophases with respect to the changes of N/P ratio and dp have been identified. The geometric features of the dendrimer molecules accommodated within the interstitial tunnels of the DNA lattices have also been revealed. The B conformation of DNA was effectively retained in the complexes in spite of the influence of the electrostatic interaction with the dendrimers. PMID- 19296684 TI - Amphiphilic polyesters derived from silylated and germylated fatty compounds. AB - New classes of amphiphilic polyesters were prepared from metallated (Si, Ge) fatty methyl ester (FAME) precursors and poly(tetramethylene oxide) glycol. Hydrosilylation of 10-undecenoic methyl ester by tetramethyldisiloxane occurred at 80 degrees C in the presence of Karstedt's catalyst, and hydrogermylation of the same FAME derivative was obtained at the same temperature under radical AIBN initiation. These diester precursors, obtained in high yields (approximately 90%), reacted with poly(tetramethylene oxide) glycol under free solvent to give silicon polymers or germanium oligomers. These condensed materials display both the characteristic of organic-inorganic hybrid materials and those of amphiphilic polymers. The nature of organometallic fragment (hydrophobicity of tetramethyldisiloxy and sterical hindrance of diphenylgermyl) was shown to influence the chemical reactivity of the polymerizable monomers and the physical properties of the resulting copolymers. The amphiphilicity of these materials provides a driving force for the formation of small objects (approximately 1 nm), making them very attractive as hybrid nanocontainers. PMID- 19296685 TI - Sequencing of single and double stranded RNA oligonucleotides by acid hydrolysis and MALDI mass spectrometry. AB - Treatment of RNA oligonucleotides with strong acids at pH 1-2 rapidly leads to hydrolysis of the phosphodiester bonds at the 5'-position of ribose. Analysis of the resulting degradation products by MALDI coupled to an Orbitrap high resolution mass spectrometer shows almost complete mass ladders from both sides of the nucleotides without interfering fragments from base losses or internal fragments. From the mass differences between adjacent peaks of a mass ladder, the sequence can be determined. Low cleavage efficiency at the termini leads to 2mers and 3mers which can be identified by MS/MS. In this way the complete sequences of different siRNA 21mer single and double strands could be verified. This simple and fast method can be applied for controlling sequences of synthetic oligomers, as well as for de-novo sequencing. Moreover, the method is applicable for localization and identification of RNA modifications as demonstrated using the examples of an oligonucleotide with phosphorothioate backbone and of one containing 2'-methoxy-ribose modifications. PMID- 19296686 TI - Current recordings of ion channel proteins immobilized on resin beads. AB - Current ion channel current measurement techniques are cumbersome, as they require many steps and much time. This is especially true when reconstituting channels into liposomes and incorporating them into lipid bilayers. Here, we report a novel method that measures ion channel current more efficiently than current methods. We applied our method to KcsA and MthK channels by binding them to cobalt affinity gel beads with histidine tags and then forming a lipid bilayer membrane on the bead. This allowed channels to incorporate into the bilayer and channel currents to be measured quickly and easily. The efficiency was such that currents could be recorded with extremely low amounts of protein. In addition, the channel direction could be determined by the histidine tag. This method has the potential to be applied to various channel proteins and channel research in general. PMID- 19296687 TI - Chemistry, physical chemistry, and uses of molecular fluorocarbon--hydrocarbon diblocks, triblocks, and related compounds--unique "apolar" components for self assembled colloid and interface engineering. PMID- 19296688 TI - Development of spin-labeled pargyline analogues as specific inhibitors of human monoamine oxidases A and B. AB - Three TEMPO-conjugated pargyline analogues (ParSL-1, ParSL-2, and ParSL-3) have been synthesized and their inhibitory properties tested for the two human monoamine oxidase isoforms (hMAOA and hMAOB). The three analogues differ in flexibility and substituent positions (para or meta) of the linkers connecting the TEMPO group to the pargyline phenyl ring. ParSL-1 contains a flexible acetamido (-CH(2)-CO-NH-) linker connecting the two moieties at the para position. In contrast, the TEMPO moieties in ParSL-2 and ParSL-3 are attached with rigid amido (-CO-NH-) linkers to the para or meta positions of the pargyline phenyl ring, respectively. These variations in conformational flexibility and substituent position are shown to have profound effects in tuning the specificities of these analogues toward the two MAO isoforms. ParSL-1 irreversibly inhibits either MAOA and MAOB, ParSL-2 inhibits only MAOB (K(i) = 15 +/- 5 microM), and ParSL-3 is found to be specific for MAOA (K(i) = 268 +/- 72 microM). These results thus provide additional insights into the role of conformational flexibility and structural properties of MAO inhibitors in tuning their isoform specificities. These active site probes have been used to determine the topological orientation of these enzymes in the mitochondrial membrane. Studies with intact mitochondria show MAOA is topologically on the cytosolic face of the outer membrane in human placenta but recombinant MAOA is situated on the opposite inner face in Pichia mitochondria. Recombinant MAOB is found to be situated on the cytosolic face of the outer membrane in Pichia mitochondria. PMID- 19296689 TI - Cytochrome C mutants for superoxide biosensors. AB - The effect of introducing positive charges (lysines) in human cytochrome c (cyt c) on the redox properties and reaction rates of cyt c with superoxide radicals was studied. The mutated forms of this electron-transfer protein are used as sensorial recognition elements for the amperometric detection of the reactive oxygen radical. The proteins were prepared by site-directed mutagenesis focusing on amino acids near the heme edge. The 11 mutants of human cyt c expressed in the course of this research have been characterized by UV-vis spectroscopy, circular dichroism, and NMR spectroscopy to verify overall structure integrity as well as axial coordination of the heme iron. The mutants are investigated voltammetrically using promoter-modified gold electrodes with respect to redox activity and formal redox potential. The rate constants for the reaction with superoxide have been determined spectrophotometrically. Four mutants show a higher reaction rate with the radical as compared to the wild type. These mutants are used for the construction of superoxide sensors based on thiol-modified gold electrodes and covalently fixed proteins. We found that the E66K mutant-based electrode has a clearly higher sensitivity in comparison with the wild-type-based sensor while retaining the high selectivity and showing a good storage stability. PMID- 19296690 TI - In situ lipidomic analysis of nonalcoholic fatty liver by cluster TOF-SIMS imaging. AB - Mass spectrometry imaging has been used to map liver biopsies of several patients suffering from nonalcoholic fatty liver disease. This steatosis is characterized by an accumulation of triacylglycerols and diacylglycerols in the liver. Using time-of-flight-secondary ion mass spectrometry (TOF-SIMS) with a bismuth cluster ion source, it has been possible to map lipids in situ at the micrometer scale and to simultaneously characterize their molecular distribution on liver sections. Accumulation of triacylglycerols, diacylglycerols, monoacylglycerols, fatty acids, with the apparition of myristic acid, together with a dramatic depletion of vitamin E and a selective macrovacuolar localization of cholesterol are observed in steatosis areas of fatty livers compared to control livers. These ion species are concentrated in small vesicles having a size of a few micrometers. Moreover, very fine differences in lipid localizations, depending on alkyl acid chain lengths of diacylglycerols and fatty acids, have been found after careful scrutiny of the ion images. Finally, TOF-SIMS has revealed lipid zonation in the normal human liver and accumulation of very similar lipids to those detected in areas of the fatty livers, which are not characterized as steatotic ones by the histological control performed on serial tissue sections. PMID- 19296691 TI - Facile synthesis of a benzindenoazepine alkaloid, bulgaramine, via samarium diiodide promoted ring expansion of an alpha-aminocarbonyl compound. AB - A novel synthetic path to a benzindenoazepine alkaloid was established by employing a samarium diiodide promoted ring expansion reaction of an alpha aminocarbonyl compound as a key reaction, in which a regioselective carbon nitrogen bond cleavage followed by ring-closing reactions occurred to give the basic ring skeleton of the target compound. Bulgaramine was synthesized from the known tetrahydroisoquinoline derivative in 5 steps in 50% overall yield. PMID- 19296692 TI - A purely lanthanide-based complex exhibiting ferromagnetic coupling and slow magnetic relaxation behavior. AB - Two lanthanide-organic frameworks, {Ln(TDA)(1.5)(H(2)O)(2)}(n) [TDA = thiophene 2,5-dicarboxylic acid anion; Ln = Gd (1), Dy (2)], were structurally and magnetically characterized. The magnetic studies revealed that the ferromagnetic coupling existed between adjacent lanthanide ions in 1 and 2, and only 2 displays slow magnetic relaxation behavior with tau(0) = 2.4 x 10(-8) s and DeltaE/k(B) = 44.2 K. To our knowledge, it is rather rare that ferromagnetic coupling and slow magnetic relaxation coexist in three-dimensional lanthanide-based frameworks. PMID- 19296693 TI - Selective monoarylation of acetate esters and aryl methyl ketones using aryl chlorides. AB - Simple, efficient procedures for the monoarylation of acetate esters and aryl methyl ketones using aryl chlorides are presented. Previously, no general method was available to ensure the highly selective monoarylation of these classes of substrates using aryl chlorides. Using palladium precatalysts recently reported by our group, these reactions are easily accomplished under mild conditions that tolerate a wide array of heterocyclic substrates. PMID- 19296694 TI - Synthesis of celecoxib analogues possessing a N-difluoromethyl-1,2-dihydropyrid-2 one 5-lipoxygenase pharmacophore: biological evaluation as dual inhibitors of cyclooxygenases and 5-lipoxygenase with anti-inflammatory activity. AB - A novel class of 1-(4-methanesulfonylphenyl and 4-aminosulfonylphenyl)-5-[4-(1 difluoromethyl-1,2-dihydropyrid-2-one)]-3-trifluoromethyl-1H-pyrazole hybrid cyclooxygenase-2 (COX-2)/5-lipoxygenase (5-LOX) inhibitory anti-inflammatory agents was designed. Replacement of the tolyl ring present in celecoxib by the N difluoromethyl-1,2-dihydropyrid-2-one moiety provided compounds showing dual selective COX-2/5-LOX inhibitory activities. 1-(4-Aminosulfonylphenyl)-5-[4-(1 difluoromethyl-1,2-dihydropyrid-2-one)]-3-trifluoromethyl-1H-pyrazole exhibited good anti-inflammatory (AI) activity (ED(50) = 27.7 mg/kg po) that compares favorably with the reference drugs celecoxib (ED(50) = 10.8 mg/kg po) and ibuprofen (ED(50) = 67.4 mg/kg po). The N-difluoromethyl-1,2-dihydropyridin-2-one moiety provides a novel 5-LOX pharmacophore for the design of cyclic hydroxamic mimetics for exploitation in the development of COX-2/5-LOX inhibitory AI drugs. PMID- 19296695 TI - Development of a novel virtual screening cascade protocol to identify potential trypanothione reductase inhibitors. AB - The implementation of a novel sequential computational approach that can be used effectively for virtual screening and identification of prospective ligands that bind to trypanothione reductase (TryR) is reported. The multistep strategy combines a ligand-based virtual screening for building an enriched library of small molecules with a docking protocol (AutoDock, X-Score) for screening against the TryR target. Compounds were ranked by an exhaustive conformational consensus scoring approach that employs a rank-by-rank strategy by combining both scoring functions. Analysis of the predicted ligand-protein interactions highlights the role of bulky quaternary amine moieties for binding affinity. The scaffold hopping (SHOP) process derived from this computational approach allowed the identification of several chemotypes, not previously reported as antiprotozoal agents, which includes dibenzothiepine, dibenzooxathiepine, dibenzodithiepine, and polycyclic cationic structures like thiaazatetracyclo-nonadeca-hexaen-3-ium. Assays measuring the inhibiting effect of these compounds on T. cruzi and T. brucei TryR confirm their potential for further rational optimization. PMID- 19296697 TI - In memory of pierre-gilles de gennes. PMID- 19296699 TI - List of collaborators and students of pierre-gilles de gennes. PMID- 19296698 TI - Biography of Pierre-Gilles de Gennes. PMID- 19296700 TI - Publications of pierre-gilles de gennes. PMID- 19296701 TI - The mechanics and fluctuation spectrum of active gels. AB - Recent experiments on molecular motor driven in vitro F-Actin networks have found anomalously large strain fluctuations at low frequency. In addition, the shear modulus of these active networks becomes as much as one hundred times larger than that of the same system in equilibrium. We develop a two-fluid model of a low density semiflexible network driven by molecular motors to explore these effects and show that, relying on only simple assumptions regarding the motor activity in the system we can quantitatively understand both the low-frequency fluctuation enhancement and the nonequilibrium stiffening of the network. These results have implications for the interpretation of microrheology in such active networks including the cytoskeleton of living cells. In addition, they may form the basis for theoretical studies of biomimetic nonequilibrium gels whose mechanical properties are tunable through the control of their nonequilibrium steady-state. PMID- 19296702 TI - Role of molecular structure on X-ray diffraction in uniaxial and biaxial phases of thermotropic liquid crystals. AB - X-ray diffraction is one of the most definitive methods to determine the structure of condensed matter phases, and it has been applied to unequivocally infer the structures of conventional calamitic and lyotropic liquid crystals. With the advent of bent-core and tetrapodic mesogens and the discovery of the biaxial nematic phase in them, the experimental results require more careful interpretation and analysis. Here, we present ab-initio calculations of X-ray diffraction patterns in the isotropic, uniaxial nematic, and biaxial nematic phases of bent-core mesogens. A simple Meier-Saupe-like molecular distribution function is employed to describe both aligned and unaligned mesophases. The distribution function is decomposed into two, polar and azimuthal, distribution functions to calculate the effect of the evolution of uniaxial and biaxial nematic orientational order. The calculations provide satisfactory semiquantitative interpretations of experimental results. The calculations presented here should provide a pathway to more refined and quantitative analysis of X-ray diffraction data from the biaxial nematic phase. PMID- 19296703 TI - Phase diagrams of semisoft nematic elastomers. AB - Nematic elastomers with locked-in anisotropy direction exhibit semisoft elastic response characterized by a plateau in the stress-strain curve in which stress does not change with strain. We calculate the global phase diagram for a minimal model, which is equivalent to one describing a nematic in crossed electric and magnetic fields, and show that semisoft behavior is associated with a broken symmetry biaxial phase and that it persists well into the supercritical regime. We also consider generalizations beyond the minimal model, particularly the neo classical model introduced by Warner, Bladon, and Terentjev, and find similar results. Our work underscores the fact that semisoft response is fundamentally a nonlinear phenomenon and that experiments on samples at zero external stress provide no information about it. PMID- 19296704 TI - The force acting on a polymer partially confined in a tube. AB - We consider the force acting on a polymer part of whose length is configurationally confined in a tube and the rest of which is free. This situation arises in many different physical contexts, including a flexible synthetic polymer partially confined in a nanopore and a stiff viral genome partially ejected from its capsid. In both cases the force acting to pull the chain molecule out of its confinement is argued to be constant once a few persistence lengths are "free"/"outside". We present Brownian dynamics simulations that confirm the constancy of the force for different chain lengths and illustrate the dependence of the force on the strength of tube confinement. Experimental results are reported for genome ejection from viral capsids, from which we estimate the pulling force to be a few tenths of a piconewton. PMID- 19296705 TI - Electric susceptibility of sodium-doped water clusters by beam deflection. AB - The electric susceptibility of neutral sodium-doped water clusters Na(H(2)O)(N), N = 6-33, was determined by beam electric deflection. The clusters behave as polarizable particles; their intensity profiles exhibit global shifts toward the high-field region without the occurrence of broadening. In the conditions of the experiment, sodium-water clusters have a "floppy" structure and hence the electric susceptibility presents both electronic and orientacional terms. Measured susceptibilities are somewhat higher than those of pure water clusters, and the contribution per water molecule is similar for both cluster types. PMID- 19296706 TI - The interpretation of sulfur K-edge XANES spectra: a case study on thiophenic and aliphatic sulfur compounds. AB - Sulfur K-edge XANES has been measured for three sulfur model compounds, dibenzothiophene, dibenzothiophene sulfone, and aliphatic sulfur (DL-methionine). The spectra have been simulated with Density Functional Theory (DFT) by using a number of methods, including the half-core-hole approximation. Dipole transition elements were calculated and the transitions were convoluted with linearly increasing Gaussian functions in the first 20 eV of the near-edge region. In the case of dibenzothiophene, relaxation of the first excited states in the presence of the core-hole gave a further improvement. The theoretical results reproduce well the features of the spectra and give insight in the relation between geometric structure and molecular orbitals. Though DL-methionine and dibenzothiophene show a similar sharp rise of the white line, their molecular levels are quite different, pointing out the difficulties in finding useful "fingerprints" in the spectra for specific compounds. PMID- 19296707 TI - Measurement of heterogeneous reaction rates during indium-mediated allylation. AB - Indium-mediated allylation provides remarkable stereo- and regioselectivity, and it proceeds easily and in high yield in aqueous solutions. In spite of its widespread use, there have been few fundamental studies of this reaction. We have developed a photomicrographic technique for measuring rates of reaction of allyl halides at indium surfaces, and we describe the mathematical model for discriminating between diffusion and kinetic control. The measurements demonstrate that this reaction is diffusion controlled, and the minimum value of the heterogeneous rate constant is 1 x 10(-3) cm s(-1). These results broaden the applicability of photomicroscopy for measuring heterogeneous rates of reactions that result in consumption of solid metals. PMID- 19296708 TI - Resonance Raman characterization of different forms of ground-state 8-bromo-7 hydroxyquinoline caged acetate in aqueous solutions. AB - The 8-bromo-7-hydroxyquinolinyl group (BHQ) is a derivative of 7-hydroxyquinoline (7-HQ) and BHQ molecules coexisting as different forms in aqueous solution. Absorption and resonance Raman spectroscopic methods were used to examine 8-bromo 7-hydroxyquinoline protected acetate (BHQ-OAc) in acetonitrile (MeCN), H(2)O/MeCN (60:40, v/v, pH 6 approximately 7), and NaOH-H(2)O/MeCN (60:40, v/v, pH 11 approximately 12) to obtain a better characterization of the forms of the ground state species of BHQ-OAc in aqueous solutions and to examine their properties. The absorption spectra of BHQ-OAc in water show no absorption bands of the tautomeric species unlike the strong band at about 400 nm observed for the tautomeric form in 7-HQ aqueous solution. The resonance Raman spectra in conjunction with Raman spectra predicted from density functional theory (DFT) calculations reveal the observation of a double Raman band system characteristic of the neutral form (the nominal C=C ring stretching, C-N stretching, and O-H bending modes at 1564 and 1607 cm(-1)) and a single Raman band diagnostic of the enol-deprotonated anionic form (the nominal C=C ring, C-N, and C-O(-) stretching modes in the 1593 cm(-1) region). These results suggest that the neutral form of BHQ-OAc is the major species in neutral aqueous solution. There is a modest increase in the amount of the anionic form and a big decrease in the amount of the tautomeric form of the molecules for BHQ-OAc compared to 7-HQ in neutral aqueous solution. The presence of the 8-bromo group and/or competitive hydrogen bonding that hinder the formation and transfer process of a BHQ-OAc-water cyclic complex may be responsible for this large substituent effect. PMID- 19296709 TI - Multichannel RRKM-TST and direct-dynamics VTST study of the reaction of hydroxyl radical with furan. AB - The kinetics and mechanism of the reaction of OH with furan have been theoretically studied. The potential energy surface for each possible pathway has been investigated by employing DFT, G3MP2, and CCSD methods. The potential energy surface consists of one hydrogen-bonded complex and two energized intermediates. Three different pathways are suggested to be possible for the title reaction. The most probable channel is the hydroxyl radical addition to the C(2) position on the furan ring to cause the ring-opening process. The two other pathways are hydrogen abstraction from one of the C(2) or C(3) position on furan and hydroxyl radical substitution at the C(2) or C(3) position on furan. Abstraction and substitution channels are minor paths at low temperature, but they become comparable with addition channels at high temperature. Addition and substitution reactions proceed via formation of two energized intermediates, Int(1) and Int(2). Multichannel RRKM-TST calculations have been carried out to calculate the individual and overall rate constants for addition and substitution reactions. Direct-dynamics canonical variational transition-state theory calculations with small curvature approximation for tunneling were carried out to study hydrogen abstraction pathways. PMID- 19296710 TI - Interaction of water highly diluted in 1-alkyl-3-methyl imidazolium ionic liquids with the PF6(-) and BF4(-) anions. AB - We have investigated water highly diluted in 1-alkyl-3-methyl imidazolium ionic liquids (ILs) with hexafluorophosphate {PF(6)(-)} and tetrafluoroborate {BF(4)( )} anions using vibrational spectroscopic measurements in the nu(OH) spectral domain of water (3600-3800 cm(-1)) and DFT calculations. The measured profiles exhibit two well-defined bands at coinciding vibrational transitions assigned with the nu(1) symmetric and nu(3) antisymmetric OH stretching modes of monodispersed water. The local organization and the vibrational spectra of water diluted in ILs have been assessed by DFT calculations (using the B3LYP functional and 6-31+G** basis set). We show that the predicted structures of water interacting (minimally) with two anions in nearly "symmetric" structures of type (A...H-O-H...A) lead to spectral features consistent with the previous spectroscopic observations as well as with those reported here. We emphasize the role of the non additive interaction forces (especially the 3-bodies electrostatic interactions) in the structural organization taking place between the cation-anion couples and for determining preferentially (A...H-O-H...A) associations of water with the anions as well as their consequences on the vibrational spectra of water. We show that the doubly hydrogen-bonded character of water in such associations leads to well-defined spectral features, which are the shifts of the nu(1) and nu(3) stretching modes of water, the separation Delta nu(13) between them (about 80 cm(-1)), and the intensity ratio estimates R = I nu(3)/I nu(1) (IR absorption and Raman). Finally, we evoke the fact that the H bond interactions of water diluted in these ILs involve a more noticeable electrostatic character than for H-bond interactions of water in usual molecular solvents. In this context, we emphasize that the appearance of the Raman band of the nu(3) mode of water originates from a significant polarization of water due to the local electrostatic fields induced by surrounding ions. PMID- 19296711 TI - Halide vacancies created by the heterogeneous reaction of OH with alkali halide single crystals. AB - The heterogeneous surface reaction of OH with dry KI(100) results in iodide vacancies in the surface lattice sites that are filled with OH to generate a stable layer of KOH. Under high-vacuum conditions, in which surface ions are not mobile, the reaction is self-passivating and generates two molecular layers of potassium hydroxide, releasing 1.6 x 10(16) iodide ions per cm(2) of surface area. Reaction rates are identical with those of NaI(100). A similar surface reaction occurs with alkali bromides (KBr(100)), albeit at a much slower rate to generate approximately one-tenth of a monolayer of KOH, whereas no observable reaction occurs with KCl(100) under the conditions of this experiment. The heterogeneous reaction of OH with alkali halides is found to be dependent solely on the identity of the halide anion and independent of the alkali metal cation with the relative reaction rates following the anion ordering, I(-) > Br(-) > Cl( ). The release of halide-containing species is expected to impact the chemistry of the marine boundary layer. PMID- 19296712 TI - Homonuclear versus heteronuclear resonance-assisted hydrogen bonds: tautomerism, aromaticity, and intramolecular hydrogen bonding in heterocyclic systems with different exocyclic proton donor/acceptor. AB - Tautomerism and resonance-assisted hydrogen bonding have been analyzed on the basis of the results of ab initio calculations at the MP2/AUG-cc-pVDZ level of theory for the series of molecules containing different heterocycles connected with resonance spacer and containing different exocyclic proton donor/acceptor atoms. It is demonstrated that the position of tautomeric equilibrium is controlled mainly by two factors: aromaticity of heterocycle, which could be different for two tautomers, and relative proton affinities of two heteroatoms forming a hydrogen bond. Replacement of exocyclic proton donor/acceptor atom results in change of an aromaticity degree of heterocycle leading to alteration of relative stability of tautomers. Comparison of structure and properties of E and Z conformers of molecules demonstrates resonance-assisted character of intramolecular hydrogen bond. Application of the NBO theory reveals that the pi component of the electron density within resonant spacer plays the primary role for determination of characteristics of hydrogen bond while sigma-skeleton only reflects the pi-polarization. An analysis of strength of intramolecular hydrogen bond using geometrical, energetic, and AIM and NBO parameters indicates that the homonuclear N...H-N hydrogen bond is considerably weaker than heteronuclear N...H O and N...H-S hydrogen bonds in the case of the XH tautomers. PMID- 19296713 TI - Dynamics and stability of individual base pairs in two homologous RNA-DNA hybrids. AB - Nuclear magnetic resonance spectroscopy and proton exchange have been used to characterize two RNA-DNA hybrids from the tR2 intrinsic transcription terminator site of phage lambda. The hybrids have the same base sequence [5' GGCGCAGGCC(T/U)(T/U)CC-3'/5'-GGAAGGCC(T/U)GCGCC-3'] but differ from each other by an interchange of DNA and RNA strands. The opening of single base pairs in the two hybrids is characterized by measuring the rates of exchange of imino protons with solvent protons as a function of the concentration of a proton acceptor (ammonia base) at 10 degrees C. The free energy change in the opening reaction provides a measure of the stability of the base pair, while the rates of opening and closing define the base pair dynamics. The results demonstrate that, within the same base sequence context, dA-rU base pairs are less stable than dT-rA base pairs. The differences in stability are enhanced when two dA-rU base pairs are located next to each other in the hybrid structure. For the G-C base pairs, the rates of opening and closing and the stability are affected by the base sequence context and by the nature of the sugar moiety attached to the guanine. The dominant feature of the base sequence is the proximity of the dA-rU base pair, which destabilizes the G-C base pair when the guanine is located on the DNA strand. Two G-C base pairs (namely, those in the fourth and 10th positions) exhibit large differences in their opening and closing rates between the two hybrids, while maintaining the same stability. These results provide the first demonstration that, for RNA-DNA hybrid structures with the same base sequence, the opening dynamics and the stability of individual base pairs are strongly influenced by the chemical nature of each strand. PMID- 19296714 TI - Interactions between small heat shock protein alpha-crystallin and galectin related interfiber protein (GRIFIN) in the ocular lens. AB - As a member of the small heat shock protein superfamily, alpha-crystallin has a chaperone-like ability to recognize and bind denatured or unfolded proteins and prevent their aggregation. Recent studies suggest that alpha-crystallin may also interact with a variety of proteins under native conditions in vitro. To identify potential binding partners for alpha-crystallin in the intact ocular lens, we conducted cross-linking studies in transgenic mouse lenses designed for overexpression of His-tagged human alphaA-crystallin. Interacting proteins were copurified with the epitope-tagged crystallin complexes and were identified by tandem mass spectrometry. This approach identified GRIFIN (galectin-related interfiber protein) as a novel binding partner. Consistent with results from cross-linking, GRIFIN subunits copurified with alpha-crystallin complexes during size exclusion chromatography of nontransgenic mouse lens extracts prepared without chemical cross-linking. Equilibrium binding to GRIFIN was studied using native alpha-crystallin isolated from calf lenses as well as oligomeric complexes reconstituted from recombinant alphaA- and alphaB-crystallin subunits. Calf lens alpha-crystallin binds GRIFIN with relatively high affinity (K(d) = 6.5 +/- 0.8 microM) at a stoichiometry of 0.25 +/- 0.01 GRIFIN monomer/alpha-crystallin subunit. The binding interaction between alpha-crystallin and GRIFIN is enhanced up to 5-fold in the presence of 3 mM ATP. These binding data support the hypothesis that GRIFIN is a novel binding partner of alpha-crystallin in the lens. PMID- 19296715 TI - Prediction of the intrinsic hydrogen bond acceptor strength of organic compounds by local molecular parameters. AB - A quantum chemical model has been developed for predicting the hydrogen bond (HB) acceptor strength of monofunctional organic compounds from electronic ground state properties of the single molecules. Local molecular parameters are used to quantify electrostatic, polarizability, and charge transfer components to hydrogen bonding, employing the ab initio and density functional theory levels HF/6-31G** and B3LYP/6-31G**. The model can handle lone pairs of intermediate and strong HB acceptor heteroatoms (N, O, S) as well as of weak HB acceptor halogens (F, Cl, Br) and includes also olefinic, alkyne, and aromatic pi-bonds as weak HB acceptor sites. The model calibration with 403 compounds and experimental values for the Abraham HB acceptor strength B yielded squared correlation coefficients r(2) around 0.95, outperforming existing fragment-based schemes. Model validation was performed applying a leave-50%-out procedure, yielding predictive squared correlation coefficients q(2) of around 0.95 for the subsets that both cover the whole chemical domain as well as (almost) the whole target value range of the data set. PMID- 19296716 TI - Rational design and 3D-pharmacophore mapping of 5'-thiourea-substituted alpha thymidine analogues as mycobacterial TMPK inhibitors. AB - Thymidine monophosphate kinase (TMPK) has emerged as an attractive target for developing inhibitors of Mycobacterium tuberculosis growth. In this study the receptor-independent (RI) 4D-QSAR formalism has been used to develop QSAR models and corresponding 3D-pharmacophores for a set of 5'-thiourea-substituted alpha thymidine inhibitors. Models were developed for the entire training set and for a subset of the training set consisting of the most potent inhibitors. The optimized (RI) 4D-QSAR models are statistically significant (r(2) = 0.90, q(2) = 0.83 entire set, r(2) = 0.86, q(2) = 0.80 high potency subset) and also possess good predictivity based on test set predictions. The most and least potent inhibitors, in their respective postulated active conformations derived from the models, were docked in the active site of the TMPK crystallographic structure. There is a solid consistency between the 3D-pharmacophore sites defined by the QSAR models and interactions with binding site residues. This model identifies new regions of the inhibitors that contain pharmacophore sites, such as the sugar pyrimidine ring structure and the region of the 5'-arylthiourea moiety. These new regions of the ligands can be further explored and possibly exploited to identify new, novel, and, perhaps, better antituberculosis inhibitors of TMPKmt. Furthermore, the 3D-pharmacophores defined by these models can be used as a starting point for future receptor-dependent antituberculosis drug design as well as to elucidate candidate sites for substituent addition to optimize ADMET properties of analog inhibitors. PMID- 19296717 TI - Upper-room ultraviolet light and negative air ionization to prevent tuberculosis transmission. AB - BACKGROUND: Institutional tuberculosis (TB) transmission is an important public health problem highlighted by the HIV/AIDS pandemic and the emergence of multidrug- and extensively drug-resistant TB. Effective TB infection control measures are urgently needed. We evaluated the efficacy of upper-room ultraviolet (UV) lights and negative air ionization for preventing airborne TB transmission using a guinea pig air-sampling model to measure the TB infectiousness of ward air. METHODS AND FINDINGS: For 535 consecutive days, exhaust air from an HIV-TB ward in Lima, Peru, was passed through three guinea pig air-sampling enclosures each housing approximately 150 guinea pigs, using a 2-d cycle. On UV-off days, ward air passed in parallel through a control animal enclosure and a similar enclosure containing negative ionizers. On UV-on days, UV lights and mixing fans were turned on in the ward, and a third animal enclosure alone received ward air. TB infection in guinea pigs was defined by monthly tuberculin skin tests. All guinea pigs underwent autopsy to test for TB disease, defined by characteristic autopsy changes or by the culture of Mycobacterium tuberculosis from organs. 35% (106/304) of guinea pigs in the control group developed TB infection, and this was reduced to 14% (43/303) by ionizers, and to 9.5% (29/307) by UV lights (both p < 0.0001 compared with the control group). TB disease was confirmed in 8.6% (26/304) of control group animals, and this was reduced to 4.3% (13/303) by ionizers, and to 3.6% (11/307) by UV lights (both p < 0.03 compared with the control group). Time-to-event analysis demonstrated that TB infection was prevented by ionizers (log-rank 27; p < 0.0001) and by UV lights (log-rank 46; p < 0.0001). Time-to-event analysis also demonstrated that TB disease was prevented by ionizers (log-rank 3.7; p = 0.055) and by UV lights (log-rank 5.4; p = 0.02). An alternative analysis using an airborne infection model demonstrated that ionizers prevented 60% of TB infection and 51% of TB disease, and that UV lights prevented 70% of TB infection and 54% of TB disease. In all analysis strategies, UV lights tended to be more protective than ionizers. CONCLUSIONS: Upper-room UV lights and negative air ionization each prevented most airborne TB transmission detectable by guinea pig air sampling. Provided there is adequate mixing of room air, upper-room UV light is an effective, low-cost intervention for use in TB infection control in high-risk clinical settings. PMID- 19296718 TI - Furious rabies after an atypical exposure. PMID- 19296719 TI - Media portrayals of suicide. PMID- 19296720 TI - Cargo and dynamin regulate clathrin-coated pit maturation. AB - Total internal reflection fluorescence microscopy (TIR-FM) has become a powerful tool for studying clathrin-mediated endocytosis. However, due to difficulties in tracking and quantifying their heterogeneous dynamic behavior, detailed analyses have been restricted to a limited number of selected clathrin-coated pits (CCPs). To identify intermediates in the formation of clathrin-coated vesicles and factors that regulate progression through these stages, we used particle-tracking software and statistical methods to establish an unbiased and complete inventory of all visible CCP trajectories. We identified three dynamically distinct CCP subpopulations: two short-lived subpopulations corresponding to aborted intermediates, and one longer-lived productive subpopulation. In a manner dependent on AP2 adaptor complexes, increasing cargo concentration significantly enhances the maturation efficiency of productive CCPs, but has only minor effects on their lifetimes. In contrast, small interfering RNA (siRNA) depletion of dynamin-2 GTPase and reintroduction of wild-type or mutant dynamin-1 revealed dynamin's role in controlling the turnover of abortive intermediates and the rate of CCP maturation. From these data, we infer the existence of an endocytic restriction or checkpoint, responsive to cargo and regulated by dynamin. PMID- 19296721 TI - Oncogenic Kras initiates leukemia in hematopoietic stem cells. AB - How oncogenes modulate the self-renewal properties of cancer-initiating cells is incompletely understood. Activating KRAS and NRAS mutations are among the most common oncogenic lesions detected in human cancer, and occur in myeloproliferative disorders (MPDs) and leukemias. We investigated the effects of expressing oncogenic Kras(G12D) from its endogenous locus on the proliferation and tumor-initiating properties of murine hematopoietic stem and progenitor cells. MPD could be initiated by Kras(G12D) expression in a highly restricted population enriched for hematopoietic stem cells (HSCs), but not in common myeloid progenitors. Kras(G12D) HSCs demonstrated a marked in vivo competitive advantage over wild-type cells. Kras(G12D) expression also increased the fraction of proliferating HSCs and reduced the overall size of this compartment. Transplanted Kras(G12D) HSCs efficiently initiated acute T-lineage leukemia/lymphoma, which was associated with secondary Notch1 mutations in thymocytes. We conclude that MPD-initiating activity is restricted to the HSC compartment in Kras(G12D) mice, and that distinct self-renewing populations with cooperating mutations emerge during cancer progression. PMID- 19296722 TI - Converging intracranial markers of conscious access. AB - We compared conscious and nonconscious processing of briefly flashed words using a visual masking procedure while recording intracranial electroencephalogram (iEEG) in ten patients. Nonconscious processing of masked words was observed in multiple cortical areas, mostly within an early time window (<300 ms), accompanied by induced gamma-band activity, but without coherent long-distance neural activity, suggesting a quickly dissipating feedforward wave. In contrast, conscious processing of unmasked words was characterized by the convergence of four distinct neurophysiological markers: sustained voltage changes, particularly in prefrontal cortex, large increases in spectral power in the gamma band, increases in long-distance phase synchrony in the beta range, and increases in long-range Granger causality. We argue that all of those measures provide distinct windows into the same distributed state of conscious processing. These results have a direct impact on current theoretical discussions concerning the neural correlates of conscious access. PMID- 19296723 TI - The implications of multiple circadian clock origins. PMID- 19296724 TI - Human cord blood transplantation in a neonatal rat model of hypoxic-ischemic brain damage: functional outcome related to neuroprotection in the striatum. AB - Human umbilical cord blood mononuclear cells (HUCB) have been shown to have a therapeutic role in different models of central nervous system (CNS) damage, including stroke. We evaluated the possible therapeutic potential of HUCB in P7 rats submitted to the Rice-Vannucci model of neonatal hypoxic-ischemic (HI) brain damage. Our results demonstrated that intraperitoneal transplantation of HUCB, 3 h after the HI insult, resulted in better performance in two developmental sensorimotor reflexes, in the first week after the injury. We also showed a neuroprotective effect in the striatum, and a decrease in the number of activated microglial cells in the cerebral cortex of treated animals. We suggest that HUCB transplantation might rescue striatal neurons from cell death after a neonatal HI injury resulting in better functional recovery. PMID- 19296725 TI - Pauses and intonational phrasing: ERP studies in 5-month-old German infants and adults. AB - In language learning, infants are faced with the challenge of decomposing continuous speech into relevant units, such as syntactic clauses and words. Within the framework of prosodic bootstrapping, behavioral studies suggest infants approach this segmentation problem by relying on prosodic information, especially on acoustically marked intonational phrase boundaries (IPBs). In the current ERP study, we investigate processing of IPBs in 5-month-old infants by varying the acoustic cues signaling the IPB. In an experiment in which pitch variation, vowel lengthening, and pause cues are present (Experiment 1), 5-month old German infants show an ERP obligatory response. This obligatory response signals lower level perceptual processing of acoustic cues that, however, disappear when no pause cue is present (Experiment 2). This suggests that infants are sensitive to sentence internal pause, a cue that is relevant for the processing of IPBs. Given that German adults show both the obligatory components and the closure positive shift, a particular ERP component known to reflect the perception of IPBs, independent of the presence of a pause cue, the results of the current ERP study indicate clear developmental differences in intonational phrase processing. The comparison of our neurophysiological data from German learning infants with behavioral data from English-learning infants furthermore suggests cross-linguistic differences in intonational phrase processing during infancy. These findings are discussed in the light of differences between the German and the English intonation systems. PMID- 19296726 TI - Real-time tracking of motor response activation and response competition in a Stroop task in young children: a lateralized readiness potential study. AB - The ability to select an appropriate motor response by resolving competition among alternative responses plays a major role in cognitive performance. fMRI studies suggest that the development of this skill is related to the maturation of the frontal cortex that underlies the improvement of motor inhibition abilities. However, fMRI cannot characterize the temporal properties of motor response competition and motor activation in general. We studied the development of the time course of resolving motor response competition. To this end, we used the lateralized readiness potential (LRP), an ERP measure, for tracking correct and incorrect motor cortex activation in children in real time. Fourteen children and 14 adults took part in an animal-size Stroop task where they selected between two animals, presented simultaneously on the computer screen, which was larger in real life. In the incongruent condition, the LRP detected stronger and longer lasting incorrect response activation in children than in adults. LRP results could explain behavioral congruency effects, the generally longer RT in children than in adults and the larger congruency effect in children than in adults. In contrast, the peak latency of ERP waves, usually associated with stimulus processing speed, could explain neither of the above effects. We conclude that the development of resolving motor response competition, relying on motor inhibition skills, is a crucial factor in child development. Our study demonstrates that the LRP is an excellent tool for studying motor activation in children. PMID- 19296727 TI - Behavioral and electrophysiological evidence of motor cortex activation related to an amputated limb: a multisensorial approach. AB - Phantom limb sensations may be linked to motor activities in the deafferented cortices of amputees, with artificial visual feedback of an amputated limb leading to enhanced phantom sensations. The present study was designed to verify if cortical motor activity related to an amputated limb can be triggered by visual input using an objective behavioral measure and with a neurophysiological correlate. Trauma amputees and normally limbed subjects showed superior performance in a mirror-drawing task when the mirror was placed sagittally (giving visual feedback of the amputated/inactive limb) compared with when it was placed frontally. Measurement of lateralized movement-related brain potentials showed that, under the lateral mirror condition, contralateral motor activity of the viewed hand was observed in both normal subjects and trauma amputees. In contrast, this activity was not observed in subjects with congenital limb absence. These findings suggest that, in traumatic amputees, motor enhancement due to visualization of the movements of the missing limb reflects the effectiveness of motor commands to the missing limb, strengthening the hypothesis of the functional survival of deafferented cortical motor areas. PMID- 19296728 TI - Neural signatures of semantic and phonemic fluency in young and old adults. AB - As we age, our ability to select and to produce words changes, yet we know little about the underlying neural substrate of word-finding difficulties in old adults. This study was designed to elucidate changes in specific frontally mediated retrieval processes involved in word-finding difficulties associated with advanced age. We implemented two overt verbal (semantic and phonemic) fluency tasks during fMRI and compared brain activity patterns of old and young adults. Performance during the phonemic task was comparable for both age groups and mirrored by strongly left-lateralized (frontal) activity patterns. On the other hand, a significant drop of performance during the semantic task in the older group was accompanied by additional right (inferior and middle) frontal activity, which was negatively correlated with performance. Moreover, the younger group recruited different subportions of the left inferior frontal gyrus for both fluency tasks, whereas the older participants failed to show this distinction. Thus, functional integrity and efficient recruitment of left frontal language areas seems to be critical for successful word retrieval in old age. PMID- 19296729 TI - A picture is worth a thousand dollars. PMID- 19296733 TI - Running for your life or running for your dinner: what drives fiber-type evolution in lizard locomotor muscles? AB - Despite its role in whole-animal performance, the adaptation of muscle physiology related to terrestrial locomotion remains underexplored. We tested evolutionary models based on predator escape and foraging strategies of lizards to assess whether fiber-type composition of a leg muscle is adaptive for behavior. The best fitting model for fast-twitch fiber-type evolution was one based on predator escape strategy, while the foraging-mode model fared poorly (Akaike Information Criterion with small sample size correction; DeltaAICc=29.7). According to the predator-escape model, lizards relying on sprints to avoid predators are predicted to have relatively higher proportions of fast glycolytic (FG) fibers (70%), while cryptic lizards are predicted to have relatively higher fast oxidative glycolytic (FOG) fiber proportions (77%). This pattern suggests an evolutionary trend toward greater FG (FOG) fiber composition among lizards that specialize in sprinting (crypsis). The best-fitting model for slow-twitch fibers had a single optimum, suggesting a common selective pressure across these lizards. The second-best model explaining slow-twitch fiber-type evolution was Brownian motion (DeltaAICc=0.80), indicating some support for neutral evolution. We find evidence suggesting that different fiber types occurring in the same muscle can evolve under different evolutionary pressures. PMID- 19296734 TI - Conflict over reproduction in an ant-plant symbiosis: why Allomerus octoarticulatus ants sterilize Cordia nodosa trees. AB - The evolutionary stability of mutualism is thought to depend on how well the fitness interests of partners are aligned. Because most ant-myrmecophyte mutualisms are persistent and horizontally transmitted, partners share an interest in growth but not in reproduction. Resources invested in reproduction are unavailable for growth, giving rise to a conflict of interest between partners. I investigated whether this explains why Allomerus octoarticulatus ants sterilize Cordia nodosa trees. Allomerus octoarticulatus nests in the hollow stem domatia of C. nodosa. Workers protect C. nodosa leaves against herbivores but destroy inflorescences. Using C. nodosa trees with Azteca ants, which do not sterilize their hosts, I cut inflorescences off trees to simulate sterilization by A. octoarticulatus. Sterilized C. nodosa grew faster than control trees, providing evidence for a trade-off between growth and reproduction. Allomerus octoarticulatus manipulates this trade-off to its advantage; sterilized trees produce more domatia and can house larger, more fecund colonies. PMID- 19296735 TI - Reproductive conflict and the costs of social status in cooperatively breeding vertebrates. AB - Conflict over reproduction is an inherent part of group living. In many social vertebrates, conflict may be reflected as allostatic load, or the costs of social status and dominance rank, which may be quantified by measuring glucocorticoid stress hormones. Here, we develop the first quantitative model of allostatic load based on the tug-of-war model of reproductive skew to generate insights into the mechanisms underlying reproductive conflict in cooperative breeders and to determine whether glucocorticoids can be used to assess conflict levels in group living vertebrates. It predicts that subordinates have higher allostatic loads than dominants under most conditions, but when body condition is lower in dominants than in subordinates, dominants experience higher allostatic load. Group structure is also important, as dominants generally have higher allostatic loads than subordinates when there is a large number of subordinates in the group, but this cost can be reduced by increasing the number of dominants, as in plural breeding societies. Using glucocorticoid data from cooperatively breeding superb starlings Lamprotornis superbus, we found empirical support for both predictions. Our model is useful for understanding how the costs of social status influence reproductive sharing, and it suggests that glucocorticoids can be used to examine reproductive conflict and cooperation in social species. PMID- 19296736 TI - Asexual fungal symbionts alter reproductive allocation and herbivory over time in their native perennial grass hosts. AB - Although most plants harbor microbial symbionts, the temporal effects of symbionts on resource allocation and herbivore resistance of perennial hosts are unknown. Neotyphodium endophytes are asexual, vertically transmitted fungal symbionts of grasses that are thought to interact mutualistically with their hosts, mainly by deterring herbivores. To test age-specific effects of Neotyphodium infection and herbivory on resource allocation, I conducted a 4-year field experiment with four genotypes of an infected perennial native grass from which the endophyte was removed and for which herbivory and water availability were controlled. In the absence of herbivory, infection increased allocation to reproductive effort in the first two growing seasons. Infected plants also flowered earlier. Herbivory increased allocation to reproduction in the first year but much more so in infected than in uninfected plants. Infected plants also had greater herbivore loads in early stages, suggesting that infected plants are more tolerant to herbivory. Asexual fungal symbionts thus fundamentally alter host resource allocation and resistance and tolerance to herbivores over time. Increased reproductive effort in early host ontogeny should benefit the symbiont by increasing transmission but perhaps at the expense of lifetime host fitness. If so, then the conventional notion of asexual endophytes as mutualistic hostages of their hosts is incorrect. PMID- 19296737 TI - Native fauna on exotic trees: phylogenetic conservatism and geographic contingency in two lineages of phytophages on two lineages of trees. AB - The relative roles of evolutionary history and geographical and ecological contingency for community assembly remain unknown. Plant species, for instance, share more phytophages with closer relatives (phylogenetic conservatism), but for exotic plants introduced to another continent, this may be overlaid by geographically contingent evolution or immigration from locally abundant plant species (mass effects). We assessed within local forests to what extent exotic trees (Douglas-fir, red oak) recruit phytophages (Coleoptera, Heteroptera) from more closely or more distantly related native plants. We found that exotics shared more phytophages with natives from the same major plant lineage (angiosperms vs. gymnosperms) than with natives from the other lineage. This was particularly true for Heteroptera, and it emphasizes the role of host specialization in phylogenetic conservatism of host use. However, for Coleoptera on Douglas-fir, mass effects were important: immigration from beech increased with increasing beech abundance. Within a plant phylum, phylogenetic proximity of exotics and natives increased phytophage similarity, primarily in younger Coleoptera clades on angiosperms, emphasizing a role of past codiversification of hosts and phytophages. Overall, phylogenetic conservatism can shape the assembly of local phytophage communities on exotic trees. Whether it outweighs geographic contingency and mass effects depends on the interplay of phylogenetic scale, local abundance of native tree species, and the biology and evolutionary history of the phytophage taxon. PMID- 19296738 TI - The brief multidimensional students' life satisfaction scale-college version. AB - OBJECTIVES: To investigate the psychometric properties of the BMSLSS-College among 723 college students. METHODS: Internal consistency estimates explored scale reliability, factor analysis explored construct validity, and known-groups validity was assessed using the National College Youth Risk Behavior Survey and Harvard School of Public Health College Alcohol Study. Criterion-related validity was explored through analyses with the CDC's health-related quality of life scale and a social isolation scale. RESULTS: Acceptable internal consistency reliability, construct, known-groups, and criterion-related validity were established. CONCLUSIONS: Findings offer preliminary support for the BMSLSS-C; it could be useful in large-scale research studies, applied screening contexts, and for program evaluation purposes toward achieving Healthy People 2010 objectives. PMID- 19296739 TI - Body weight and perceived social pressure to exercise at a health club. AB - OBJECTIVE: To determine the social pressure overweight (OW, n=1183) and normal weight (NW, n=550) adults perceive regarding health club exercise. METHODS: A 12 item survey, based on the theory of reasoned action and theory of planned behavior, measured subjective norm toward exercising at a health club 30 min, twice a week, for the next month. RESULTS: OW feel more social pressure to exercise at a club than do NW, but motivation to comply is the same between groups. CONCLUSIONS: Exercise interventions for OW should capitalize on individual perceived social pressures to exercise by designing strategies to increase motivation to comply with these pressures. PMID- 19296740 TI - Reliability and validity of the alcohol consequences expectations scale. AB - OBJECTIVES: To examine the reliability and validity of a new measure of alcohol outcome expectations for college students, the Alcohol Consequences Expectations Scale (ACES). METHODS: College students (N=169) completed the ACES and several other measures. RESULTS: Results support the existence of 5 internally consistent subscales. Additionally, the ACES is associated with conceptually similar measures and self-reported drinking behavior. CONCLUSIONS: This study supports the reliability of the ACES and its subscales and provides preliminary evidence of construct and criterion-related validity. Pending further investigation, this scale may be used to inform the development of alcohol abuse prevention programs on college campuses. PMID- 19296741 TI - Validity of random-digit-dialing in recruiting controls in a case-control study. AB - OBJECTIVES: To evaluate whether the participating controls represented the underlying population in a colorectal cancer case-control study in a geographically isolated North American population. METHODS: The characteristics of the study controls recruited through Random-digit-dialing (RDD) were compared with those in the corresponding target population estimates. RESULTS: Participating controls were more likely to have higher levels of education and income and were less likely to be smokers. CONCLUSION: Study controls recruited through RDD tend to have higher socioeconomic status, which may lead to overestimation of a number of risk factors in this study. PMID- 19296742 TI - Self-efficacy moderates the mediation of intentions into behavior via plans. AB - OBJECTIVE: To examine the putative moderating role of self-efficacy in the intention-planning-behavior relationship. METHODS: In N=812 individuals, intentions (independent variable) were assessed at baseline, whereas action plans (mediator), self-efficacy (moderator), and physical activity (dependent variable) were measured again 4 weeks later. We examined a moderated-mediation model. RESULTS: Self-efficacy moderates the mediation process: the strength of the mediated effect increased along with levels of self-efficacy. The results remain valid after accounting for baseline physical activity. CONCLUSIONS: For plans to mediate the intention-behavior relation, people must hold sufficiently high levels of self-efficacy. If they lack self-efficacy, planning may be in vain. PMID- 19296744 TI - Validation of self-report on smoking among university students in Korea. AB - OBJECTIVE: To validate the self-reported smoking status of Korean university students. METHODS: Subjects included 322 Korean university in Korea, who participated in an annual health screening. Data on smoking were collected through a self-reported questionnaire and urine test. The data were analyzed by the McNemar test. RESULTS: In the consistency test for males, the results from the questionnaire and the urine test were in agreement, but for females, there was a significant difference between the results reported by the 2 tests (P=.001). CONCLUSIONS: The findings showed that Korean female university students considerably underreported their smoking behavior. PMID- 19296745 TI - Health literacy skills in rural and urban populations. AB - OBJECTIVE: To determine whether health literacy is lower in rural populations. METHOD: We analyzed health, prose, document, and quantitative literacy from the National Assessment of Adult Literacy study. Metropolitan Statistical Area designated participants as rural or urban. RESULTS: Rural populations had lower literacy levels for all literacy types (P<0.001 for each). After adjusting for known confounders, there was no longer a difference in health or prose literacy (P>0.05). However, rural populations had higher document (P=0.04) and quantitative (P=0.01) literacy. CONCLUSION: Health literacy is lower in the rural population although this difference is explained by known confounders. PMID- 19296743 TI - Adopting a plant-based diet minimally increased food costs in WHEL Study. AB - OBJECTIVE: To assess the cost of adopting a plant-based diet. METHODS: Breast cancer survivors randomized to dietary intervention (n=1109) or comparison (n=1145) group; baseline and 12-month data on diet and grocery costs. RESULTS: At baseline, both groups reported similar food costs and dietary intake. At 12 months, only the intervention group changed their diet (vegetable-fruit: 6.3 to 8.9 serv/d.; fiber: 21.6 to 29.8 g/d; fat: 28.2 to 22.3% of E). The intervention change was associated with a significant increase of $1.22/ person/week (multivariate model, P=0.027). CONCLUSIONS: A major change to a plant-based diet was associated with a minimal increase in grocery costs. PMID- 19296746 TI - Attitudes and sociocultural factors influencing vaginal douching behavior among English-speaking Latinas. AB - OBJECTIVES: To describe the practice/correlates of douching among a national sample of English-speaking Latinas. METHODS: Three hundred twenty-five (325) computer-assisted telephone interviews were completed. Analyses compare ever douchers to never douchers and identify correlates of douching. RESULTS: Of respondents, 50% had ever douched; 15% douched at least monthly. Douching practices among respondents were similar to those of other groups. Age at sexual debut was the strongest correlate of ever douching. Regular douching significantly increased as the perceived benefits of douching and the number of sources recommending douching increased. CONCLUSIONS: Interventions to prevent douching and to support cessation should be tailored to characteristics and perceptions of Latinas. Targeting secondary audiences may prove valuable. PMID- 19296747 TI - Young adult smoking behavior: implications for future population health. AB - OBJECTIVE: To qualitatively project the future health burden of tobacco from present-day young adult smoking behavior. METHODS: Population surveys in California (2002) and nationally (1978-80, 2001-03). RESULTS: In 2002, 40% of California young adult smokers were nondaily smokers, 24% had quit at some time for >or=6 months, 45% said they smoked less now than previously, and 68% thought they would quit within 5 years. Interest in quitting was high, and most were actively engaged in the smoking cessation process. Young adult smoking behavior changed from 1978-80 to 2001-03. CONCLUSION: The future health burden from smoking will be less for the current generation of young adults. PMID- 19296749 TI - Factor retention in EFA: strategies for health behavior researchers. AB - OBJECTIVE: To provide readers with information on various techniques to improve future health behavior EFA research. METHODS: Data from a health education questionnaire development study are analyzed to make the discussion of techniques concrete. RESULTS: There are numerous strategies to use when making a factor retention decision. CONCLUSIONS: The informed health-behavior researcher should use sophisticated factor retention techniques during EFAs. PMID- 19296748 TI - Recruitment and retention of Latino children in a lifestyle intervention. AB - OBJECTIVE: To describe promising recruitment and retention strategies for transient Latino populations, assisting investigators who work with this population in their research design and implementation. METHODS: Strategies in recruitment and retention from a year-long intervention in children and their families are described. RESULTS: Of the 159 families who agreed to participate in the program, 123 parent-child dyads were enrolled. Retention rates were 59% in the control group, 67% in the intervention group. CONCLUSION: Building and maintaining relationships between a consistent study team contact and participants promote strong recruitment and retention outcomes. Barriers such as child care and transportation must be minimized. PMID- 19296750 TI - Alternative high school students: prevalence and correlates of overweight. AB - OBJECTIVE: To determine prevalence and correlates of overweight among adolescents attending alternative high schools (AHS). METHODS: AHS students (n=145) from 6 schools completed surveys and anthropometric measures. Cross-sectional associations were assessed using mixed model multivariate logistic regression. RESULTS: Among students, 42% were overweight, with female, low-income and racial/ethnic minorities disproportionately affected. The odds of being overweight were significantly associated with self-described health status, social support, family meals, intentions to eat healthy at school, and use of healthy weight management strategies. CONCLUSIONS: Overweight was prevalent among AHS students and requires further study. Correlates provide targets for future research. PMID- 19296751 TI - Protective behaviors and high-risk drinking among entering college freshmen. AB - OBJECTIVES: To explore the use of protective behaviors to reduce risks associated with alcohol consumption among adolescents during the summer preceding college enrollment. METHODS: Survey data were collected in fall 2006 and 2007 that assessed demographic characteristics, drinking behaviors, and use of protective behaviors in the 3 months preceding the survey. RESULTS: Female participants reported using 4 out of 10 protective behaviors more often than did males, and using protective behaviors was significantly related to fewer negative drinking related consequences. CONCLUSIONS: Findings highlight potential benefits of using protective behaviors and the need to promote effective behaviors. PMID- 19296753 TI - Vascular dysfunction in Type 2 diabetes: emerging targets for therapy. PMID- 19296754 TI - Helping the circulatory system heal itself: manipulating kinin signaling to promote neovascularization. PMID- 19296755 TI - The Dieter test. PMID- 19296756 TI - Power lines and implantable rhythm devices. PMID- 19296759 TI - Efficacy and safety of olmesartan medoxomil alone and in combination with hydrochlorothiazide. AB - In at least 50% of patients with hypertension, particularly those with stage 2 hypertension, combination drug therapy is required to achieve the currently recommended blood pressure (BP) goals. It is likely that future hypertension guidelines will recommend lowering BP beyond the currently recommended systolic/diastolic BP goals of lower than 140/90 mmHg for all patients with hypertension and lower than 130/80 mmHg for patients with diabetes or chronic kidney disease. In a series of clinical trials, the combination of olmesartan medoxomil, a well-established angiotensin receptor blocker, and hydrochlorothiazide, a thiazide diuretic, has produced greater reductions in systolic BP and diastolic BP and greater proportions of patients achieving BP goals than monotherapy with either component. Additionally, the increased efficacy resulting from the combination with hydrochlorothiazide does not appear to significantly affect the tolerability profile of olmesartan medoxomil. This article provides a comprehensive evaluation of the chemistry, clinical efficacy, safety and tolerability of this combination and discusses its role in the management of hypertension. PMID- 19296760 TI - Fondaparinux in the treatment of acute coronary syndromes: evidence from OASIS 5 and 6. AB - Anticoagulant therapy is widely used for the management of acute coronary syndromes. In order to optimize patient outcomes, anticoagulants should ideally combine high antithrombotic efficacy with a low risk of bleeding. Intravenous unfractionated heparin has been in clinical use for more than 50 years and reduces the risk of recurrent ischemic events in patients with acute coronary syndromes but at the cost of increased bleeding. Enoxaparin, compared with intravenous unfractionated heparin, further reduces the risk of ischemic events but also increases bleeding. Neither of these approaches has been shown to reduce mortality. The synthetic parenteral Factor Xa inhibitor, fondaparinux, is highly effective for the prevention and treatment of venous thromboembolic disease in medical and surgical patients. The Organization for the Assessment of Strategies for Ischemic Syndromes (OASIS) 5 and 6 trials evaluated the efficacy and safety of fondaparinux in more than 32,000 patients with non-ST elevation acute coronary syndromes or ST elevation myocardial infarction. This clinical trial report discusses the findings of these two pivotal trials. PMID- 19296761 TI - Cardio-renal protection with aliskiren, a direct renin inhibitor, in the ASPIRE HIGHER program. AB - Aliskiren is the first in a new class of orally effective direct renin inhibitors. Initial results of clinical efficacy studies have demonstrated at least equivalent or superior blood pressure-lowering efficacy when compared with existing drugs, and a favorable side-effect profile either as monotherapy or as a component of combination therapy. This report aims to introduce and provide a critical appraisal of the initial results of the ASPIRE HIGHER program, evaluating the potential cardio-renal protective effects of aliskiren. PMID- 19296762 TI - Multidisciplinary approach for circulatory support in patients with advanced heart failure. AB - Despite the efficacy of heart-failure (HF) therapy, patients may still decompensate and require hospitalization. In addition to the gap between guidelines and clinical practice, this highlights the need for a multidisciplinary approach to the management of HF. The third European Mechanical Circulatory Support Summit presented the latest trials and trends in conservative therapy of end-stage HF (ESHF) and the alternative options over a ten-session program. The meeting covered topics such as epidemiological trends, different HF registries and guidelines for ESHF therapy. In addition to databases and HF trials, the presenters discussed the integration of multiple biomarkers in HF stratification, biological solution and embryonic stem cells for HF therapy, up to-date myocardial recovery and the surgical aspects of ESHF treatment. Therapeutic options following short-term left ventricular support were discussed, such as: which patients should receive a ventricular assist device (VAD) and when; what the perfect window for implantation of a VAD is; bridge-to-bridge and device selection; VAD-weaning criteria; left ventricular unloading, patient device matching (short-term device), cardiac transplantation following short-term support. There was a device update, presenting Circulite, Abiomed Impella, Levacor, Levitronix, VentrAssist, Heartmate II, DuraHeart and Heartware. Finally, the International VAD registry, European VAD registry and The International Society for Heart & Lung Transplantation Mechanical Circulatory Device database were also discussed. Herein, a commentary is givenon some of the interesting topics that indicate the importance of the community, the physician and the patient's awareness of HF, the utilization of the updated guidelines and landmark studies and registries, as well as recent evidence-based modalities in the management of ESHF. PMID- 19296763 TI - Perils of weight loss: the advantage of being obese in patients with heart failure. AB - Evaluation of: Pocock SJ, McMurray JJV, Dobson J et al. Weight loss and mortality risk in patients with chronic heart failure in the Candesartan in Heart failure: Assessment of Reduction in Mortality and morbidity (CHARM) programme. Eur. Heart J. 29, 2641-2650 (2008). Among patients with heart failure, the usual association between obesity and reduced survival appears to be turned on its head. Increasing numbers of studies appear to confirm the apparent protective effect of the 'obesity paradox' in heart failure. The adverse effects of weight loss and cachexia have been recognized among patients with chronic disease. The paper under evaluation clarifies and offers further insight into the impact of BMI and weight loss on prognosis in a large sample of heart failure patients. The results confirm and extend previous knowledge. They raise many questions, not only regarding the underlying pathophysiology that leads to this counterintuitive relationship observed in the obese heart failure group, but also as to how we should best approach and advise these patients on weight management in day-to-day practice. PMID- 19296764 TI - Diabetes and hypertension: which is the best approach? AB - Evaluation of: Ostergren J, Poulter NR, Sever PS et al. The Anglo-Scandinavian Cardiac Outcomes Trial: blood pressure-lowering limb: effects in patients with Type II diabetes. J. Hypertens. 26, 2103-2111 (2008). Although blood pressure control is crucial to improve cardiovascular prognosis in patients with diabetes and hypertension, less than 10% of these patients attain blood pressure goals. European guidelines recommend that combined therapy should be used as first-line treatment in this population. The next step in the treatment of these patients is to determine which combination is preferable for each situation. The results of the Anglo-Scandinavian Cardiac Outcomes Trial demonstrated the benefits of amlodipine-based treatment on the incidence of total cardiovascular events and procedures, compared with atenolol-based treatment. In this article the potential explanations of these results are discussed. PMID- 19296765 TI - Metabolic syndrome, cardiovascular risk and screening for subclinical atherosclerosis. AB - The metabolic syndrome is a clustering of risk factors known to promote or increase the risk of diabetes development and subsequent cardiovascular disease. Screening for subclinical atherosclerosis using new imaging technologies or novel biomarkers could help to further risk-stratify patients with metabolic syndrome. In particular, noninvasive imaging of carotid intima-media thickness and coronary artery calcium scoring seem to have promising prognostic value in identifying patients at high risk. Early identification could lead to improved patient or physician adherence to risk-reducing behaviors or interventions and improve clinical outcomes. PMID- 19296766 TI - The emerging obesity problem in Latin America. AB - Obesity and overweight prevalence is increasing rapidly in Latin America. This increase has been attributed to lifestyle changes occurring in recent decades related to rapid socioeconomic development, including a more Westernized diet, physical inactivity, urbanization, rural-urban migration and some maternal-fetal factors. In addition, genetic factors may have a role, inducing a higher predisposition to accumulate abdominal fat and develop metabolic syndrome. This increase in obesity and metabolic syndrome is leading to higher morbidity and mortality due to Type 2 diabetes and cardiovascular disease. In a few poor countries, obesity coexists with undernutrition, making the situation even more difficult. Global intervention, from both governments and nongovernmental organizations, is necessary. They must play an active role, monitoring the food market and facilitating community-based initiatives that promote a healthy lifestyle. PMID- 19296768 TI - Pathophysiology of coronary artery disease: the case for multiparametric imaging. AB - Interventions to treat coronary artery disease are available but they must be targeted at the correct individuals (and indeed lesions), in order to gain maximal benefit with the minimal adverse effects. Coronary contrast angiography is not able to provide all the information required for the assessment of the effects of artery disease. Other imaging modalities are of growing importance as they can reduce radiation exposure and invasiveness of screening, as well as providing important extra information. The ideal 'multiparametric' imaging technique would assess anatomy, viability and lesion activity in a single quick scan. Currently, MRI is the technology closest to achieving this ideal, although the existing technology still has some limitations. This review discusses the currently available techniques for the imaging of coronary anatomy and of myocardial viability, and considers their benefits and limitations. We also discuss the developing field of imaging molecularly targeted to active coronary lesions. Finally we provide a 5-year view of the current and likely future optimal imaging strategies. PMID- 19296769 TI - Management of atrial fibrillation in patients with heart failure: from drug therapy to ablation. AB - Atrial fibrillation (AF) and heart failure are the two modern epidemics of cardiovascular disease. They are commonly encountered together and either condition predisposes to the other. Patients with both AF and heart failure carry a poor prognosis; therefore, the management of this group of patients should be addressed aggressively. In the context of AF management in patients with heart failure, the pharmacologic rhythm control strategy is not superior to the rate control strategy. Due to the adverse effects of anti-arrhythmic agents, the rate control should be considered as a primary approach. The long-term data regarding the efficacy and safety of catheter-based ablation therapy in the patients with heart failure are limited; hence, the catheter ablation of AF should be reserved for the patients for whom the drug therapy is inadequate or undesirable. Ultimately, the patient's symptoms and the tolerance for the particular treatment should be taken into account for the management of AF in patients with heart failure. PMID- 19296772 TI - Role of thrombectomy in acute myocardial infarction. AB - Acute ST elevation myocardial infarction results from atherosclerotic plaque rupture with subsequent thrombus formation, leading to complete or near complete occlusion of an epicardial coronary artery. Minimization of the mechanical obstruction from this thrombus remains the main goal of therapy in ST elevation myocardial infarction. Primary percutaneous coronary intervention for an ST elevation myocardial infarction appears to be the preferred mode of revascularization over thrombolytic therapy if the door-to-balloon time target of 90 min is achievable. The idea of reducing the thrombus burden with the use of devices as an adjunct to percutaneous coronary intervention is an attractive one. Several thrombectomy devices have been studied in randomized clinical trials, but no definitive conclusions have emerged, owing to conflicting results and variable clinical end points. This article intends to shed further light on the potential role of thrombectomy in the treatment of acute myocardial infarction. PMID- 19296773 TI - Epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) nasal colonization among patients and healthcare workers in a Portuguese hospital: a pre-intervention study toward the control of MRSA. AB - This two-year study investigated the epidemiology of nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) among patients and healthcare workers (HCWs) in two wards with a high frequency of MRSA isolation, at Hospital Geral de Santo Antonio (HGSA), Portugal. Three point-prevalence surveys per year were carried out. A case-control approach was used to identify potential risk factors associated with MRSA carriage among patients. Incidence rates and risk factors of MRSA carriage among HCWs who were negative at the baseline observation were estimated. Prevalence of MRSA carriage among 276 patients screened was 5.1%. Admission to HGSA or attendance to the Diabetic Foot Outpatient Unit (DFOU) of HGSA within the past 12 months, and previous MRSA isolation were significant risk factors for MRSA carriage. Among HCWs (n = 126), the prevalence of MRSA carriage was 4.8% and the incidence rate was 61/1000 person-years. Nurses and nurse aids were the HCW categories with the highest risk of becoming colonized with MRSA over time (p = 0.01). One HCW chronically colonized was detected. Molecular typing revealed a clonal identity for isolates recovered from patients and HCWs of the same wards, with 88.6% of isolates belonging to the EMRSA-15 (ST22-MRSA IV) clone. PMID- 19296775 TI - Evidence-based public health: a fundamental concept for public health practice. AB - Despite the many accomplishments of public health, a greater attention to evidence-based approaches is warranted. This article reviews the concepts of evidence-based public health (EBPH), on which formal discourse originated about a decade ago. Key components of EBPH include making decisions on the basis of the best available scientific evidence, using data and information systems systematically, applying program-planning frameworks, engaging the community in decision making, conducting sound evaluation, and disseminating what is learned. Three types of evidence have been presented on the causes of diseases and the magnitude of risk factors, the relative impact of specific interventions, and how and under which contextual conditions interventions were implemented. Analytic tools (e.g., systematic reviews, economic evaluation) can be useful in accelerating the uptake of EBPH. Challenges and opportunities (e.g., political issues, training needs) for disseminating EBPH are reviewed. The concepts of EBPH outlined in this article hold promise to better bridge evidence and practice. PMID- 19296774 TI - Adaptive designs for randomized trials in public health. AB - In this article, we present a discussion of two general ways in which the traditional randomized trial can be modified or adapted in response to the data being collected. We use the term adaptive design to refer to a trial in which characteristics of the study itself, such as the proportion assigned to active intervention versus control, change during the trial in response to data being collected. The term adaptive sequence of trials refers to a decision-making process that fundamentally informs the conceptualization and conduct of each new trial with the results of previous trials. Our discussion below investigates the utility of these two types of adaptations for public health evaluations. Examples are provided to illustrate how adaptation can be used in practice. From these case studies, we discuss whether such evaluations can or should be analyzed as if they were formal randomized trials, and we discuss practical as well as ethical issues arising in the conduct of these new-generation trials. PMID- 19296776 TI - Health communication in the Latino community: issues and approaches. AB - With reference to the Communication-Persuasion model, we describe various research issues and challenges when considering the health of Latinos, and implications for designing and evaluating health communication and behavior change efforts in this population. Latinos, collectively the nation's largest minority group, vary substantially in terms of socioeconomic and legal status, their country of origin and the extent of ongoing contact with that country, their region of residence within the United States, their generation status and levels of acculturation, and psychosocial factors. Health communication efforts with Latinos need to focus on family, cultural traditions, and collectivism while attending to acculturation, language, generation and national origin. The most extensive intervention topic in Latino health promotion has been the application of the lay health advisor model. This and other fundamental communication approaches, as well as audience and population characteristics, need to be considered within the context of dynamic and complex societal changes. PMID- 19296777 TI - The delivery of public health interventions via the Internet: actualizing their potential. AB - The Internet increasingly serves as a platform for the delivery of public health interventions. The efficacy of Internet interventions has been demonstrated across a wide range of conditions. Much more work remains, however, to enhance the potential for broad population dissemination of Internet interventions. In this article, we examine the effectiveness of Internet interventions, with particular attention to their dissemination potential. We discuss several considerations (characterizing reach rates, minimizing attrition, promoting Web site utilization, use of tailored messaging and social networking) that may improve the implementation of Internet interventions and their associated outcomes. We review factors that may influence the adoption of Internet interventions in a range of potential dissemination settings. Finally, we present several recommendations for future research that highlight the potential importance of better understanding intervention reach, developing consensus regarding Web site usage metrics, and more broadly integrating Web 2.0 functionality. PMID- 19296778 TI - Extreme makeover: Transformation of the veterans health care system. AB - The veterans health care system administered by the U.S. Department of Veterans Affairs (VA) was established after World War I to provide health care for veterans who suffered from conditions related to their military service. It has grown to be the nation's largest integrated health care system. As the system grew, a number of factors contributed to its becoming increasingly dysfunctional. By the mid-1990s, VA health care was widely criticized for providing fragmented and disjointed care of unpredictable and irregular quality, which was expensive, difficult to access, and insensitive to individual needs. Between 1995 and 1999, the VA health care system was reengineered, focusing especially on management accountability, care coordination, quality improvement, resource allocation, and information management. Numerous systemic changes were implemented, producing dramatically improved quality, service, and operational efficiency. VA health care is now considered among the best in America, and the VA transformation is viewed as a model for health care reform. PMID- 19296779 TI - Quality-based financial incentives in health care: can we improve quality by paying for it? AB - This article asks whether financial incentives can improve the quality of health care. A conceptual framework drawn from microeconomics, agency theory, behavioral economics, and cognitive psychology motivates a set of propositions about incentive effects on clinical quality. These propositions are evaluated through a synthesis of extant peer-reviewed empirical evidence. Comprehensive financial incentives--balancing rewards and penalties; blending structure, process, and outcome measures; emphasizing continuous, absolute performance standards; tailoring the size of incremental rewards to increasing marginal costs of quality improvement; and assuring certainty, frequency, and sustainability of incentive payoffs--offer the prospect of significantly enhancing quality beyond the modest impacts of prevailing pay-for-performance (P4P) programs. Such organizational innovations as the primary care medical home and accountable health care organizations are expected to catalyze more powerful quality incentive models: risk- and quality-adjusted capitation, episode of care payments, and enhanced fee for-service payments for quality dimensions (e.g., prevention) most amenable to piece-rate delivery. PMID- 19296780 TI - The contribution of hospitals and health care systems to community health. AB - This article reviews evidence on hospitals' and health systems' impacts on community health improvement. We begin with an overview of the history of community benefit and then discuss the lack of a widely accepted definition and measurement of community benefit activities as well as the expectations and accountability of tax-exempt not-for-profit hospitals and health systems in community initiatives. We highlight the approaches of two systems and identify strategic, cultural, technical, and structural challenges associated with increasing community benefit and health-improvement activities. We conclude by offering recommendations for policy and practice. PMID- 19296781 TI - Quality of prescribing decision support in primary care: still a work in progress. PMID- 19296782 TI - Cannabis use in remote Indigenous communities in Australia: endemic yet neglected. PMID- 19296783 TI - When and how to welcome government to the bedroom. PMID- 19296784 TI - Perinatal outcomes after assisted reproductive technology treatment in Australia and New Zealand: single versus double embryo transfer. AB - OBJECTIVE: To compare the perinatal outcomes of babies conceived by single embryo transfer (SET) with those conceived by double embryo transfer (DET). DESIGN, SETTING AND PARTICIPANTS: A retrospective population-based study of embryo transfer cycles in Australia and New Zealand between 2002 and 2006, using data from the Australia and New Zealand Assisted Reproduction Database. MAIN OUTCOME MEASURES: Proportion of SET procedures; comparison of SET and DET procedures with respect to multiple births, low birthweight (LBW), preterm birth and fetal death. RESULTS: The proportion of SET procedures has increased from 28.4% in 2002 to 32.0% in 2003, 40.5% in 2004, 48.2% in 2005 and 56.9% in 2006. The multiple birth rate for all babies conceived by SET (4.0%) was 10 times lower than for those conceived by DET (39.1%) (P < 0.01). The average birthweight for all liveborn babies conceived by SET (3290 g) was higher than for those conceived by DET (2934 g) (P < 0.01). The preterm birth rate of all DET-conceived babies (30.3%) was higher than for SET-conceived babies (12.3%) (adjusted odds ratio [AOR], 3.19 [95% CI, 3.01-3.38]). All babies conceived by DET were more likely to be stillborn than those conceived by SET (AOR, 1.49 [95% CI, 1.21-1.82]). Singletons conceived by DET were more likely to be born preterm than singletons conceived by SET (AOR, 1.13 [95% CI, 1.05-1.22]). Liveborn singletons conceived by DET were 15% more likely to have LBW than liveborn singletons conceived by SET (AOR, 1.15 [95% CI, 1.05-1.26]). There was no significant difference in fetal death rate between DET- and SET-conceived singletons. CONCLUSION: The increase in proportion of SET procedures has resulted in a lower rate of multiple births and in better perinatal outcomes in Australian and New Zealand assisted reproduction programs. PMID- 19296785 TI - The impact of the Baby Bonus payment in New South Wales: who is having "one for the country"? AB - OBJECTIVE: To assess the change in birth rates, both overall and in age, parity, socioeconomic and geographical subgroups of the population, after the introduction of the Baby Bonus payment in Australia on 1 July 2004. DESIGN AND SETTING: Population-based study using New South Wales birth records and Australian Bureau of Statistics population estimates for the period 1 January 1997 - 31 December 2006. PARTICIPANTS: All 853 606 women aged 15-44 years with a pregnancy resulting in a birth at > or = 20 weeks' gestation or a baby > or = 400 g birthweight. MAIN OUTCOME MEASURE: Change in birth rate in 2005 and 2006 compared with the trend in birth rates before the introduction of the Baby Bonus. RESULTS: The crude annual birth rate showed a downward trend from 1997 to 2004; after 2004 this trend reversed with a sharp increase in 2005 and a further increase in 2006. All age-specific birth rates increased after 2004, with the greatest increase in birth rate, relative to the trend before the Baby Bonus, being seen in teenagers. Rates of first births were not significantly affected by the bonus; however, rates of third or subsequent births increased across all age, socioeconomic and geographical subgroups. CONCLUSIONS: In the first 2 years after the introduction of the Baby Bonus, birth rates increased, especially among women having a third or subsequent birth. This could represent an increase in family size and/or a change in the timing of births. PMID- 19296786 TI - Could the Baby Bonus be a bonus for babies? AB - Closing the gap in life expectancy between Indigenous and non-Indigenous Australians needs to start in the womb. Rates of perinatal mortality, preterm birth and low birthweight are two to three times greater among the babies of Indigenous women than among those of non-Indigenous women; low birthweight predisposes infants to greater risks of chronic illness in later life. Indigenous women in Australia tend to present for antenatal care later in pregnancy than do non-Indigenous women. There are many barriers for Indigenous women seeking to access antenatal care - geographical, social, cultural, financial and in some cases a lack of service provision. Many of these problems are being addressed within the public health system and by Indigenous community-controlled health services. However, more needs to be done. While antenatal care cannot solve all medical and social problems, commencing such care as early as possible in pregnancy has the potential to improve maternal health and hence pregnancy outcomes. Changes in the way the government Baby Bonus is paid to new mothers could act as an incentive not only to service providers but also to women themselves to initiate antenatal care in the first trimester of pregnancy. Such a system has been well established for many years in France. Any changes to the Baby Bonus scheme should provide incentives and not be punitive in nature. PMID- 19296787 TI - Sperm removal and dead or dying patients: a dilemma for emergency departments and intensive care units. AB - An unexpected consequence of the increase in the use of fertility treatment is that emergency department and intensive care doctors are receiving requests from wives (actual or de facto) of dying or recently deceased men for sperm removal. Legislation in all states and territories regulates removal of sperm from a dying man and, provided that lawful consent is obtained, a doctor can harvest sperm. In several states, including Victoria, harvested sperm cannot be used in a fertilisation procedure without the man's consent, and debate surrounds the issue of consent and how it can be proved. Recent Victorian Law Reform Commission recommendations attempt to streamline the law to make a man's consent the cornerstone of decision making for both harvesting and subsequent use of sperm. PMID- 19296788 TI - Oocyte freezing: timely reproductive insurance? AB - Cryopreservation of unfertilised oocytes for later use in initiating pregnancy is now a viable technology, with acceptable pregnancy rates (over 20% per thaw cycle). Oocyte cryopreservation used as a form of insurance against "social" (age related) infertility can improve the lifetime chance of pregnancy in women who defer pregnancy into their late 30s or early 40s. We report two pregnancies using oocytes that were frozen for social rather than medical reasons, as part of a larger series of nine pregnancies using cryopreserved oocytes. Use of oocytes harvested and frozen from women aged under 35 years may more than double the chance of pregnancy for a 41-year-old woman. The disadvantages of oocyte freezing for social infertility reasons include cost, the usual risks associated with in vitro fertilisation, and the lack of a guarantee of eventual pregnancy. PMID- 19296789 TI - Infertility Treatment Act or forced sterilisation program?: comment. PMID- 19296790 TI - Quality of drug interaction alerts in prescribing and dispensing software. AB - OBJECTIVE: To investigate the quality of drug interaction decision support in selected prescribing and dispensing software systems, and to compare this information with that found in a range of reference sources. DESIGN AND SETTING: A comparative study, conducted between June 2006 and February 2007, of the support provided for making decisions about 20 major and 20 minor drug interactions in six prescribing and three dispensing software systems used in primary care in Australia. Five electronic reference sources were evaluated for comparison. MAIN OUTCOME MEASURES: Sensitivity, specificity and quality of information; for major interactions: whether information on clinical effects, timeframe and pharmacological mechanism was included, whether management advice was helpful, and succinctness. RESULTS: Six of the nine software systems had a sensitivity rate > or = 90%, detecting most of the major interactions. Only 3/9 systems had a specificity rate of > or = 80%, with other systems providing inappropriate or unhelpful alerts for many minor interactions. Only 2/9 systems provided adequate information about clinical effects for more than half the major drug interactions, and 1/9 provided useful management advice for more than half of these. The reference sources had high sensitivity and in general provided more comprehensive clinical information than the software systems. CONCLUSIONS: Drug interaction decision support in commonly used prescribing and dispensing software has significant shortcomings. PMID- 19296791 TI - Supraventricular tachycardia. AB - Supraventricular tachycardia (SVT) is a common cardiac rhythm disturbance; it usually presents with recurrent episodes of tachycardia, which often increase in frequency and severity with time. Although SVT is usually not life-threatening, many patients suffer recurrent symptoms that have a major impact on their quality of life. The uncertain and sporadic nature of episodes of tachycardia can cause considerable anxiety - many patients curtail their lifestyle as a result, and many prefer curative treatment. SVT often terminates before presentation, and episodes may be erroneously attributed to anxiety. Sudden-onset, rapid, regular palpitations characterise SVT and, in most patients, a diagnosis can be made with a high degree of certainty from patient history alone. Repeated attempts at electrocardiographic documentation of the arrhythmia may be unnecessary. Treatment of SVT may not be necessary when the episodes are infrequent and self terminating, and produce minimal symptoms. When episodes of tachycardia occur frequently, are prolonged or are associated with symptoms that affect quality of life, catheter ablation is the first choice of treatment; it is a low-risk procedure with a high success rate. Long-term preventive pharmacotherapy is an alternative approach in some patients. PMID- 19296792 TI - Spontaneous chylothorax in a 2-year-old child. AB - A previously well 2-year-old girl presented with acute respiratory distress. After multiple investigations she was diagnosed with spontaneous chylothorax, attributed to strenuous vomiting. To our knowledge, this is the second reported case of spontaneous chylothorax occurring after the neonatal period. PMID- 19296793 TI - Dengue and climate change in Australia: predictions for the future should incorporate knowledge from the past. AB - Dengue transmission in Australia is currently restricted to Queensland, where the vector mosquito Aedes aegypti is established. Locally acquired infections have been reported only from urban areas in the north-east of the state, where the vector is most abundant. Considerable attention has been drawn to the potential impact of climate change on dengue distribution within Australia, with projections for substantial rises in incidence and distribution associated with increasing temperatures. However, historical data show that much of Australia has previously sustained both the vector mosquito and dengue viruses. Although current vector distribution is restricted to Queensland, the area inhabited by A. aegypti is larger than the disease-transmission areas, and is not restricted by temperature (or vector-control programs); thus, it is unlikely that rising temperatures alone will bring increased vector or virus distribution. Factors likely to be important to dengue and vector distribution in the future include increased dengue activity in Asian and Pacific nations that would raise rates of virus importation by travellers, importation of vectors via international ports to regions without A. aegypti, higher rates of domestic collection and storage of water that would provide habitat in urban areas, and growing human populations in northern Australia. Past and recent successful control initiatives in Australia lend support to the idea that well resourced and functioning surveillance programs, and effective public health intervention capabilities, are essential to counter threats from dengue and other mosquito-borne diseases. Models projecting future activity of dengue (or other vector-borne disease) with climate change should carefully consider the local historical and contemporary data on the ecology and distribution of the vector and local virus transmission. PMID- 19296794 TI - Identifying existing health care services that do not provide value for money. AB - Health systems can be improved appreciably by making them more efficient and accountable, and enhancing the quality of care, without necessarily requiring additional resources. Australia, like other nations, cannot escape making difficult health care choices in the context of resource scarcity, and the challenge of delivering quality care, informed by best available evidence, to an ageing population with multiple comorbidities. An opportunity exists for a cost saving or cost-neutral agenda of reallocation of resources within the existing health budget, through reducing the use of existing health care interventions that offer little or no benefit relative to the cost of their public subsidy. This would allow reallocation of funding towards interventions that are more cost effective, maximising health gain. Criteria based on those developed for health technology assessment (HTA) might facilitate the systematic and transparent identification of existing, potentially ineffective practices on which to prioritise candidates for assessment as to their cost-effectiveness. The process could be jointly funded by all relevant stakeholders but centrally administered, with HTA groups resourced to undertake identification and assessment and to liaise with clinicians, consumers and funding stakeholders. PMID- 19296795 TI - Oral white lesions: pitfalls of diagnosis. AB - General practitioners are often the first point of contact for patients with oral white lesions, which represent a wide spectrum of diagnoses of varying seriousness. Some clinical features are classical and others overlap between different diagnoses; they should be correlated with patient history, and sometimes other investigations, for diagnosis. Leukoplakia is a clinical term, and is a diagnosis of exclusion with no histopathological connotation. It has been redefined to describe a predominantly white lesion with premalignant potential. Patients with lesions that are potentially malignant should be referred to an oral medicine specialist or oral maxillofacial surgeon for systematic management. PMID- 19296796 TI - Mycobacterium chelonae infection in a tattoo site. PMID- 19296797 TI - SMS text messaging for contact follow-up in invasive meningococcal disease. PMID- 19296798 TI - The rise and fall of suicide in New South Wales. PMID- 19296799 TI - Recent increases in mumps incidence in Australia: the "forgotten" age group in the 1998 Australian Measles Control Campaign. PMID- 19296800 TI - High levels of confusion for cholesterol awareness campaigns. PMID- 19296801 TI - University chairs of radiology. PMID- 19296802 TI - Rediscovering university teaching hospitals for Australia. PMID- 19296803 TI - Fetal alcohol syndrome and fetal alcohol spectrum disorder in Indigenous schoolchildren. PMID- 19296804 TI - Impact of an educational intervention on general practitioners' skills in cognitive behavioural strategies. PMID- 19296805 TI - Impeding the supply of expertise in Australian health care: actions of the Australian and New Zealand College of Anaesthetists. PMID- 19296806 TI - After the fires: looking to the future using the lessons from the past. PMID- 19296807 TI - Water recycling -- forwards or backwards for public health? PMID- 19296808 TI - The "alcopops" tax: heading in the right direction. PMID- 19296809 TI - Incidence and survival after acute myocardial infarction in Indigenous and non Indigenous people in the Northern Territory, 1992-2004. AB - OBJECTIVE: To estimate the incidence and survival rates of acute myocardial infarction (AMI) for Northern Territory Indigenous and non-Indigenous populations. DESIGN AND PARTICIPANTS: Retrospective cohort study for all new AMI cases recorded in hospital inpatient data or registered as an ischaemic heart disease (IHD) death between 1992 and 2004. MAIN OUTCOME MEASURES: Population based incidence and survival rates by age, sex, Indigenous status, remoteness of residence and year of diagnosis. RESULTS: Over the 13-year study period, the incidence of AMI increased 60% in the NT Indigenous population (incidence rate ratio [IRR], 1.04; 95% CI, 1.02-1.06), but decreased 20% in the non-Indigenous population (IRR, 0.98; 95% CI, 0.97-1.00). Over the same period, there was an improvement in all-cases survival (ie, survival with and without hospital admission) for the NT Indigenous population due to a reduction in deaths both pre hospital and after hospital admission (death rates reduced by 56% and 50%, respectively). The non-Indigenous all-cases death rate was reduced by 29% as a consequence of improved survival after hospital admission; there was no significant change in pre-hospital survival in this population. Important factors that affected outcome in all people after AMI were sex (better survival for women), age (survival declined with increasing age), remoteness (worse outcomes for non-Indigenous residents of remote areas), year of diagnosis and Indigenous status (hazard ratio, 1.44; 95% CI, 1.21-1.70). CONCLUSIONS: Our results show that the increasing IHD mortality in the NT Indigenous population is a consequence of a rise in AMI incidence, while at the same time there has been some improvement in Indigenous AMI survival rates. The simultaneous decrease in IHD mortality in NT non-Indigenous people was a result of reduced AMI incidence and improved survival after AMI in those admitted to hospital. Our results inform population-specific strategies for a systemwide response to AMI management. PMID- 19296810 TI - Natural history of chronic kidney disease in Australian Indigenous and non Indigenous children: a 4-year population-based follow-up study. AB - OBJECTIVE: To describe the natural history and risk of early chronic kidney disease (CKD) in Indigenous Australian populations. DESIGN, SETTING AND PARTICIPANTS: A prospective cohort of 2266 Aboriginal and non-Aboriginal children enrolled from primary schools throughout New South Wales from February 2002 to June 2004 and followed for 4 years. MAIN OUTCOME MEASURES: Urinalysis, height, weight, blood pressure, birthweight and sociodemographic status at baseline and 2 and 4-year follow-up; CKD risk factors: haematuria, albuminuria, obesity, and systolic and diastolic hypertension. RESULTS: 2266 children (55% Aboriginal; 51% male; mean age, 8.9 years [SD, 2.0 years]) were enrolled at baseline. 1432 children (63%) were retested at 2-year follow-up, and 1506 children (67%) at 4 year follow-up. Prevalence of baseline CKD risk factors was frequent (2%-7%), but most abnormalities were transient. Besides persistent obesity (5.0%), persistence of CKD risk factors at final follow-up was low: haematuria (1.9%), albuminuria (2.4%), systolic hypertension (1.5%) and diastolic hypertension (0.2%). There was no difference in prevalence of persistent CKD risk factors between Aboriginal and non-Aboriginal children. CONCLUSIONS: Over 4 years of follow-up, Indigenous Australian children had no increased risk for early evidence of CKD. More than 70% of baseline risk factors were transient, and persistent risk factors were uncommon. Our findings suggest the increased risk for end-stage kidney disease seen in Indigenous adults is not yet manifest in these schoolchildren, and may be potentially preventable. PMID- 19296811 TI - Patterns of mortality in Indigenous adults in the Northern Territory, 1998-2003: are people living in more remote areas worse off? AB - OBJECTIVE: To quantify Indigenous mortality in the Northern Territory by remoteness of residence. DESIGN, SETTING AND PARTICIPANTS: Australian Bureau of Statistics mortality data were used to compare rates of death from chronic disease in the NT Indigenous population with rates in the general Australian population over the period 1998-2003. Rates were evaluated by categories of remoteness based on the Accessibility/Remoteness Index of Australia: outer regional areas (ORAs), remote areas (RAs) and very remote areas (VRAs). MAIN OUTCOME MEASURES: Mortality from cardiovascular disease, diabetes and renal disease; standardised mortality ratios (SMRs); percentage change in annual death rates; changes in mortality between 1998-2000 and 2001-2003. RESULTS: In 1998 2000, SMRs for all-cause mortality were 285% in ORAs, 875% in RAs and 214% in VRAs. In 2001-2003, corresponding SMRs were 325%, 731% and 208%. For the period 1998-2003, percentage changes in annual all-cause mortality were 4.4% (95% CI, 2.2%, 11.5%) in ORAs, -5.3% (95% CI, -9.6%, -0.8%) in RAs, and 1.1% (95% CI, 7.2%, 11.3%) in VRAs. In 2001-2003, compared with 1998-2000, changes in the number of Indigenous deaths were +35 in ORAs, -37 in RAs and +32 in VRAs. Similar patterns were observed for cardiovascular mortality. CONCLUSIONS: Compared with mortality in the general Australian population, Indigenous mortality was up to nine times higher in RAs, three times higher in ORAs and two times higher in VRAs. The fact that rates were lowest in VRAs runs contrary to claims that increasing remoteness is associated with poorer health status. Despite the high death rate in RAs, there was a downward trend in mortality in RAs over the study period. This was partly attributable to a fall in the absolute number of deaths. PMID- 19296812 TI - Are babies getting bigger? An analysis of birthweight trends in New South Wales, 1990-2005. AB - OBJECTIVE: To determine whether the proportion of babies born large for gestational age (LGA) in New South Wales has increased, and to identify possible reasons for any increase. DESIGN AND SETTING: Population-based study using data obtained from the NSW Midwives Data Collection, a legislated surveillance system of all births in NSW. PARTICIPANTS: All 1 273 924 live-born singletons delivered at term (> or = 37 complete weeks' gestation) in NSW from 1990 to 2005. MAIN OUTCOME MEASURES: LGA, defined as > 90th centile for sex and gestational age using 1991-1994 Australian centile charts; maternal factors associated with LGA were assessed using logistic regression. RESULTS: The proportion of babies born LGA increased from 9.2% to 10.8% (18% increase) for male infants and from 9.1% to 11.0% (21% increase) for female infants. The mean birthweight increased by 23 g for boys and 25 g for girls over the study period. Increasing maternal age, higher rates of gestational diabetes and a decline in smoking contributed significantly to these increases, but did not fully explain them. CONCLUSIONS: There is an increasing trend in the proportion of babies born LGA, which is only partly attributable to decreasing maternal smoking, increasing maternal age and increasing gestational diabetes. PMID- 19296813 TI - Calcium and bone health: position statement for the Australian and New Zealand Bone and Mineral Society, Osteoporosis Australia and the Endocrine Society of Australia. AB - This position statement was prepared by the Working Group of the Australian and New Zealand Bone and Mineral Society and Osteoporosis Australia. The final statement was endorsed by the Endocrine Society of Australia. Currently, the balance of evidence remains in favour of fracture prevention from combined calcium and vitamin D supplementation in elderly men and women. Adequate vitamin D status is essential for active calcium absorption in the gut and for bone development and remodelling. In adults with a baseline calcium intake of 500-900 mg/day, increasing or supplementing this intake by a further 500-1000 mg/day has a beneficial effect on bone mineral density. Calcium intake significantly above the recommended level is unlikely to achieve additional benefit for bone health. PMID- 19296814 TI - The benefits of oestrogen following menopause: why hormone replacement therapy should be offered to postmenopausal women. AB - Recently, two major epidemiological studies found that hormone replacement therapy (HRT) in postmenopausal women increased the risk of breast cancer. One of the studies also found that HRT increased the risk of cardiovascular disease and thrombosis. As a consequence, women were advised to cease this therapy. However, detailed analysis of these studies suggests that the conclusions may be erroneous. Other studies suggest that the timing of initiation of HRT for healthy women is critical to achieving a beneficial outcome. When begun within 5 years of menopause in healthy women, oestrogen-based HRT results in far greater benefits than adverse outcomes. There is substantial objective evidence that the benefits of HRT include: Reduced distressing symptoms of menopause. Reduced risk of osteoporotic fractures, dementia and colorectal cancer. Improved wellbeing, quality of life; improved vaginal epithelium, sexual enjoyment and bladder capacity. Improved cardiovascular system, with reduced myocardial ischaemia and cardiovascular-related death. Increased longevity. The adverse effects of HRT include: Oral HRT doubles the risk of thromboembolism. HRT promotes growth of pre existing breast cancer. PMID- 19296815 TI - Non-inferiority trials: determining whether alternative treatments are good enough. AB - New treatments that are potentially as effective as existing treatments are increasingly being developed, some of which may be preferred because of lower cost, fewer side effects, easier administration or less harm. Non-inferiority trials attempt to establish whether or not a new treatment -- drug or non-drug -- is no worse than an established treatment for which efficacy has been determined in placebo-controlled trials. Critical issues in the design and conduct of non inferiority trials include: defining the acceptable margin of adverse events that, if exceeded, will render the new treatment inferior to the standard treatment (the non-inferiority margin); calculating the sample size needed to demonstrate non-inferiority; assessing the robustness of results in terms of absolute versus relative effects, intention-to-treat versus per-protocol analyses, one-sided versus two-sided statistical tests, and observed versus expected event rates for standard treatment; evaluating all relevant outcomes, including harm; and stating conclusions that are consistent with aims and results. Many non-inferiority trials fail to meet basic quality criteria, report biased and misleading conclusions, and are unduly influenced by commercial sponsors, with some commentators going so far as labelling them unethical. Clinicians and trial investigators need to exercise caution when interpreting results of non-inferiority trials which, because they lack a placebo group, can only provide an indirect assessment of the efficacy of a new treatment compared with an existing standard, and where the choice of non-inferiority margin can be highly subjective. PMID- 19296816 TI - Subclavian stenosis causing angina after coronary artery bypass grafting. PMID- 19296817 TI - Valsalva retinopathy induced by vigorous nightclub dancing. PMID- 19296818 TI - Clinical, electrophysiological and genetic features of a large Australian family with paramyotonia congenita. AB - A 32-year-old woman with a 4-year history of multiple sclerosis presented with persistent clawing of the right hand. History revealed that she and five family members had lifelong symptoms of paradoxical myotonia (impaired relaxation of muscles following muscle contraction), exacerbated by cold. The family was diagnosed with paramyotonia congenita, based on neurophysiological and genetic studies. To our knowledge, this is the first report of an Australian family with paramyotonia congenita. PMID- 19296819 TI - Homicide and rates of renal transplantation in the United States and Australia. PMID- 19296820 TI - Management of kidney stone disease in New South Wales. PMID- 19296821 TI - Can tuberculosis mimic cancer? PMID- 19296822 TI - Hepatosplenic T-cell lymphoma following infliximab therapy for Crohn's disease. PMID- 19296823 TI - Salt intake and health in the Australian population. PMID- 19296824 TI - Peer physical examination: time to revisit? PMID- 19296825 TI - Misdiagnosis of acute eye diseases by primary health care providers: incidence and implications. PMID- 19296826 TI - Standards for health care: a necessary but unknown quantity. PMID- 19296827 TI - Eicosapentaenoic acid stimulates AMP-activated protein kinase and increases visfatin secretion in cultured murine adipocytes. AB - Visfatin is an adipokine highly expressed in visceral AT (adipose tissue) of humans and rodents, the production of which seems to be dysregulated in excessive fat accumulation and conditions of insulin resistance. EPA (eicosapentaenoic acid), an n-3 PUFA (polyunsaturated fatty acid), has been demonstrated to exert beneficial effects in obesity and insulin resistance conditions, which have been further linked to its reported ability to modulate adipokine production by adipocytes. TNF-alpha (tumour necrosis factor-alpha) is a pro-inflammatory cytokine whose production is increased in obesity and is involved in the development of insulin resistance. Control of adipokine production by some insulin-sensitizing compounds has been associated with the stimulation of AMPK (AMP-activated protein kinase). The aim of the present study was to examine in vitro the effects of EPA on visfatin production and the potential involvement of AMPK both in the absence or presence of TNF-alpha. Treatment with the pro inflammatory cytokine TNF-alpha (1 ng/ml) did not modify visfatin gene expression and protein secretion in primary cultured rat adipocytes. However, treatment of these primary adipocytes with EPA (200 mumol/l) for 24 h significantly increased visfatin secretion (P<0.001) and mRNA gene expression (P<0.05). Moreover, the stimulatory effect of EPA on visfatin secretion was prevented by treatment with the AMPK inhibitor Compound C, but not with the PI3K (phosphoinositide 3-kinase) inhibitor LY294002. Similar results were observed in 3T3-L1 adipocytes. Moreover, EPA strongly stimulated AMPK phosphorylation alone or in combination with TNF alpha in 3T3-L1 adipocytes and pre-adipocytes. The results of the present study suggest that the stimulatory action of EPA on visfatin production involves AMPK activation in adipocytes. PMID- 19296828 TI - Two pathways for cysteine biosynthesis in Leishmania major. AB - Genome mining and biochemical analyses have shown that Leishmania major possesses two pathways for cysteine synthesis--the de novo biosynthesis pathway comprising SAT (serine acetyltransferase) and CS (cysteine synthase) and the RTS (reverse trans-sulfuration) pathway comprising CBS (cystathionine beta-synthase) and CGL (cystathionine gamma-lyase). The LmjCS (L. major CS) is similar to the type A CSs of bacteria and catalyses the synthesis of cysteine using O-acetylserine and sulfide with Kms of 17.5 and 0.13 mM respectively. LmjCS can use sulfide provided by the action of MST (mercaptopyruvate sulfurtransferase) on 3-MP (3 mercaptopyruvate). LmjCS forms a bi-enzyme complex with Leishmania SAT (and Arabidopsis SAT), with residues Lys222, His226 and Lys227 of LmjCS being involved in the complex formation. LmjCBS (L. major CBS) catalyses the synthesis of cystathionine from homocysteine, but, unlike mammalian CBS, also has high cysteine synthase activity (but with the Km for sulfide being 10.7 mM). In contrast, LmjCS does not have CBS activity. CS was up-regulated when promastigotes were grown in medium with limited availability of sulfur amino acids. Exogenous methionine stimulated growth under these conditions and also the levels of intracellular cysteine, glutathione and trypanothione, whereas cysteine had no effect on growth or the intracellular cysteine levels, correlating with the low rate of transport of cysteine into the cell. These results suggest that cysteine is generated endogenously by promastigotes of Leishmania. The absence of CS from mammals and the clear differences between CBS of mammals and Leishmania suggest that each of the parasite enzymes could be a viable drug target. PMID- 19296829 TI - Haemolytic anaemia and alterations in hepatic iron metabolism in aged mice lacking Cu,Zn-superoxide dismutase. AB - The continuous recycling of haem iron following phagocytosis and catabolism of senescent and damaged red blood cells by macrophages is a crucial process in the maintenance of systemic iron homoeostasis. However, little is known about macrophage iron handling in haemolytic states resulting from a deficiency in antioxidant defences. Our observations indicate that the recently described chronic, but moderate regenerative, haemolytic anaemia of aged SOD1 (superoxide dismutase 1)-knockout mice is associated with red blood cell modifications and sensitivity to both intra- and extra-vascular haemolysis. In the present study, we have characterized the molecular pathways of iron turnover in the liver of Sod1-deficient mice. Despite iron accumulation in liver macrophages, namely Kupffer cells, we did not measure any significant change in non-haem liver iron. Interestingly, in Kupffer cells, expression of the rate-limiting enzyme in haem degradation, haem oxygenase-1, and expression of the iron exporter ferroportin were both up-regulated, whereas the hepcidin mRNA level in the liver was decreased in Sod1-/- mice. These results suggest that concerted changes in the hepatic expression of iron- and haem-related genes in response to haemolytic anaemia in Sod1-/- mice act to reduce toxic iron accumulation in the liver and respond to the needs of erythropoiesis. PMID- 19296830 TI - Frankincense oil derived from Boswellia carteri induces tumor cell specific cytotoxicity. AB - BACKGROUND: Originating from Africa, India, and the Middle East, frankincense oil has been important both socially and economically as an ingredient in incense and perfumes for thousands of years. Frankincense oil is prepared from aromatic hardened gum resins obtained by tapping Boswellia trees. One of the main components of frankincense oil is boswellic acid, a component known to have anti neoplastic properties. The goal of this study was to evaluate frankincense oil for its anti-tumor activity and signaling pathways in bladder cancer cells. METHODS: Frankincense oil-induced cell viability was investigated in human bladder cancer J82 cells and immortalized normal bladder urothelial UROtsa cells. Temporal regulation of frankincense oil-activated gene expression in bladder cancer cells was identified by microarray and bioinformatics analysis. RESULTS: Within a range of concentration, frankincense oil suppressed cell viability in bladder transitional carcinoma J82 cells but not in UROtsa cells. Comprehensive gene expression analysis confirmed that frankincense oil activates genes that are responsible for cell cycle arrest, cell growth suppression, and apoptosis in J82 cells. However, frankincense oil-induced cell death in J82 cells did not result in DNA fragmentation, a hallmark of apoptosis. CONCLUSION: Frankincense oil appears to distinguish cancerous from normal bladder cells and suppress cancer cell viability. Microarray and bioinformatics analysis proposed multiple pathways that can be activated by frankincense oil to induce bladder cancer cell death. Frankincense oil might represent an alternative intravesical agent for bladder cancer treatment. PMID- 19296831 TI - The health-related quality of life in rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis: a comparison with a selected sample of healthy people. AB - BACKGROUND: The health-related quality of life (HRQL) is an important indicator of the burden of musculoskeletal disease. The Medical Outcome Study Short-Term 36 (SF-36) is the most used tool that evaluates HRQL as a subjective perception about psychological and physical limitations due to an underlying illness. The purpose of this study was to compare the HRQL scores among patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA) and ankylosing spondylitis (AS) and a selected sample of health people and determine their relationship with measures of clinical condition. METHODS: 799 patients (469 with RA, 164 with AS, 65 with axial PsA and 101 with peripheral PsA) accepted the invitation to participate. 1579 healthy controls were used for the comparison. We calculated scores for the eight SF-36 subscales, the Physical Component Summary (PCS) score, and the Mental Component Summary (MCS) score, according to published algorithms. Disease-related characteristics included disease duration, comorbidity, a measure for disease activity and for radiographic damage. The presence of comorbidity was ascertained through patient's self-reports by the Self-Administered Comorbidity Questionnaire (SCQ). Comparison were performed with respect to sex and age, and s scores were calculated for comparison with the norm. Multivariate analyses were used to assess the relationship between HRQL and radiographic damage, disease activity, and socio-demographic data. RESULTS: The four inflammatory rheumatic diseases (IRD), compared to controls, significantly impaired all eight health concepts of the SF-36 (p < 0.0001) in both component PCS and MCS scores (p < 0.0001). Overall, the dimensions typically affected were physical functioning, limitations due to physical function, and bodily pain. The disease with the worst HRQL for those dimensions was RA. The multivariate analyses revealed that the physical component was influenced by a high disease activity and comorbidity. The severity of psoriatic lesions was associated with poor mental functioning in patients with PsA. CONCLUSION: Chronic IRD have a clearly detrimental effect on the HRQL in both sex and in age groups, and physical domain is more impaired than mental and social ones. PMID- 19296833 TI - Theoretical basis for reducing time-lines to the determination of positive Mycobacterium tuberculosis cultures using thymidylate kinase (TMK) assays. AB - BACKGROUND: In vitro culture of pathogens on growth media forms a "pillar" for both infectious disease diagnosis and drug sensitivity profiling. Conventional cultures of Mycobacterium tuberculosis (M.tb) on Lowenstein Jensen (LJ) medium, however, take over two months to yield observable growth, thereby delaying diagnosis and appropriate intervention. Since DNA duplication during interphase precedes microbial division, "para-DNA synthesis assays" could be used to predict impending microbial growth. Mycobacterial thymidylate kinase (TMKmyc) is a phosphotransferase critical for the synthesis of the thymidine triphosphate precursor necessary for M.tb DNA synthesis. Assays based on high-affinity detection of secretory TMKmyc levels in culture using specific antibodies are considered. The aim of this study was to define algorithms for predicting positive TB cultures using antibody-based assays of TMKmyc levels in vitro. METHODS AND RESULTS: Systems and chemical biology were used to derive parallel correlation of "M.tb growth curves" with "TMKmyc curves" theoretically in four different scenarios, showing that changes in TMKmyc levels in culture would in each case be predictive of M.tb growth through a simple quadratic curvature, |tmk| = at2+ bt + c, consistent with the "S" pattern of microbial growth curves. Two drug resistance profiling scenarios are offered: isoniazid (INH) resistance and sensitivity. In the INH resistance scenario, it is shown that despite the presence of optimal doses of INH in LJ to stop M.tb proliferation, bacilli grow and the resulting phenotypic growth changes in colonies/units are predictable through the TMKmyc assay. According to our current model, the areas under TMKmyc curves (AUC, calculated as the integral integral(at2+ bt + c)dt or approximately 1/3 at3+ 1/2 bt2+ct) could directly reveal the extent of prevailing drug resistance and thereby aid decisions about the usefulness of a resisted drug in devising "salvage combinations" within resource-limited settings, where second line TB chemotherapy options are limited. CONCLUSION: TMKmyc assays may be useful for reducing the time-lines to positive identification of Mycobacterium tuberculosis (M.tb) cultures, thereby accelerating disease diagnosis and drug resistance profiling. Incorporating "chemiluminiscent or fluorescent" strategies may enable "photo-detection of TMKmyc changes" and hence automation of the entire assay. PMID- 19296832 TI - Multiple factors interact to produce responses resembling spectrum of human disease in Campylobacter jejuni infected C57BL/6 IL-10-/- mice. AB - BACKGROUND: Campylobacter jejuni infection produces a spectrum of clinical presentations in humans--including asymptomatic carriage, watery diarrhea, and bloody diarrhea--and has been epidemiologically associated with subsequent autoimmune neuropathies. This microorganism is genetically variable and possesses genetic mechanisms that may contribute to variability in nature. However, relationships between genetic variation in the pathogen and variation in disease manifestation in the host are not understood. We took a comparative experimental approach to explore differences among different C. jejuni strains and studied the effect of diet on disease manifestation in an interleukin-10 deficient mouse model. RESULTS: In the comparative study, C57BL/6 interleukin-10-/- mice were infected with seven genetically distinct C. jejuni strains. Four strains colonized the mice and caused disease; one colonized with no disease; two did not colonize. A DNA:DNA microarray comparison of the strain that colonized mice without disease to C. jejuni 11168 that caused disease revealed that putative virulence determinants, including loci encoding surface structures known to be involved in C. jejuni pathogenesis, differed from or were absent in the strain that did not cause disease. In the experimental study, the five colonizing strains were passaged four times in mice. For three strains, serial passage produced increased incidence and degree of pathology and decreased time to develop pathology; disease shifted from watery to bloody diarrhea. Mice kept on an ~6% fat diet or switched from an approximately 12% fat diet to an approximately 6% fat diet just before infection with a non-adapted strain also exhibited increased incidence and severity of disease and decreased time to develop disease, although the effects of diet were only statistically significant in one experiment. CONCLUSION: C. jejuni strain genetic background and adaptation of the strain to the host by serial passage contribute to differences in disease manifestations of C. jejuni infection in C57BL/6 IL-10-/- mice; differences in environmental factors such as diet may also affect disease manifestation. These results in mice reflect the spectrum of clinical presentations of C. jejuni gastroenteritis in humans and contribute to usefulness of the model in studying human disease. PMID- 19296834 TI - Effect of praziquantel treatment of Schistosoma mansoni during pregnancy on intensity of infection and antibody responses to schistosome antigens: results of a randomised, placebo-controlled trial. AB - BACKGROUND: Praziquantel treatment of schistosomiasis during pregnancy was only recommended in 2002; hence the effects of treatment during pregnancy are not fully known. We have therefore evaluated the effects on infection intensity and the immunological effects of praziquantel treatment against Schistosoma mansoni during pregnancy, compared with treatment after delivery. METHODS: A nested cohort of 387 Schistosoma mansoni infected women was recruited within a larger trial of de-worming during pregnancy. Women were randomised to receive praziquantel or placebo during pregnancy. All women were treated after delivery. Infection intensity after treatment was assessed by a single Kato-Katz examination of stool samples with duplicate slides and categorised as undetected, light (1-99 eggs per gram (epg)), moderate (100-399 epg) or heavy (>or=400 epg). Antibodies against S. mansoni worm and egg antigens were measured by ELISA. Results were compared between women first treated during pregnancy and women first treated after delivery. RESULTS: At enrollment, 252 (65.1%) of the women had light infection (median (IQR) epg: 35 (11, 59)), 75 (19.3%) moderate (median (IQR) epg: 179(131, 227)) and 60 (15.5%) had heavy infection (median (IQR) epg: 749 (521, 1169)) with S. mansoni. At six weeks after praziquantel treatment during pregnancy S. mansoni infection was not detectable in 81.9% of the women and prevalence and intensity had decreased to 11.8% light, 4.7% moderate and 1.6% heavy a similar reduction when compared with those first treated after delivery (undetected (88.5%), light (10.6%), moderate (0.9%) and heavy (0%), p = 0.16). Parasite specific antibody levels were lower during pregnancy than after delivery. Praziquantel treatment during pregnancy boosted anti-worm IgG isotypes and to a lesser extent IgE, but these boosts were less pronounced than in women whose treatment was delayed until after delivery. Praziquantel had limited effects on antibodies against egg antigens. CONCLUSION: S mansoni antigen specific antibody levels and praziquantel-induced boosts in antibody levels were broadly suppressed during pregnancy, but this was not associated with major reduction in the efficacy of praziquantel. Long-term implications of these findings in relation to resistance to re-infection remain to be explored. PMID- 19296836 TI - Alterations in the glutathione metabolism could be implicated in the ischemia induced small intestinal cell damage in horses. AB - BACKGROUND: Colic could be accompanied by changes in the morphology and physiology of organs and tissues, such as the intestine. This process might be, at least in part, due to the accumulation of oxidative damage induced by reactive oxygen (ROS) and reactive nitrogen species (RNS), secondary to intestinal ischemia. Glutathione (GSH), being the major intracellular thiol, provides protection against oxidative injury. The aim of this study was to investigate whether ischemia-induced intestinal injury could be related with alterations in GSH metabolism. RESULTS: Ischemia induced a significant increase in lipid hydroperoxides, nitric oxide and carbon monoxide, and a reduction in reduced glutathione, and adenosine triphosphate (ATP) content, as well as in methionine adenosyl-transferase and methyl-transferase activities. CONCLUSION: Our results suggest that ischemia induces harmful effects on equine small intestine, probably due to an increase in oxidative damage and proinflammatory molecules. This effect could be mediated, at least in part, by impairment in glutathione metabolism. PMID- 19296835 TI - Oral phosphatidylcholine pretreatment alleviates the signs of experimental rheumatoid arthritis. AB - INTRODUCTION: Phosphatidylcholine and phosphatidylcholine-derived metabolites exhibit anti-inflammatory properties in various stress conditions. We hypothesized that dietary phosphatidylcholine may potentially function as an anti inflammatory substance and may decrease inflammatory activation in a chronic murine model of rheumatoid arthritis (collagen-induced arthritis). METHODS: The experiments were performed on male DBA1/J mice. In groups 1 to 3 (n = 10 each), collagen-induced arthritis was induced by administration of bovine collagen II. In group 2 the animals were fed ad libitum with phosphatidylcholine-enriched diet as a pretreatment, while the animals of group 3 received this nourishment as a therapy, after the onset of the disease. The severity of the disease and inflammation-linked hyperalgesia were evaluated with semiquantitative scoring systems, while the venular leukocyte-endothelial cell interactions and functional capillary density were assessed by means of in vivo fluorescence microscopy of the synovial tissue. Additionally, the mRNA expressions of cannabinoid receptors 1 and 2, TNFalpha and endothelial and inducible nitric oxide synthase were determined, and classical histological analysis was performed. RESULTS: Phosphatidylcholine pretreatment reduced the collagen-induced arthritis-induced hypersensitivity, and decreased the number of leukocyte-endothelial cell interactions and the extent of functional capillary density as compared with those of group 1. It also ameliorated the tissue damage and decreased inducible nitric oxide synthase expression. The expressions of the cannabinoid receptors and TNFalpha were not influenced by the phosphatidylcholine intake. Phosphatidylcholine-enriched food administrated as therapy failed to evoke the aforementioned changes, apart from the reduction of the inducible nitric oxide synthase expression. CONCLUSIONS: Phosphatidylcholine-enriched food as pretreatment, but not as therapy, appears to exert beneficial effects on the morphological, functional and microcirculatory characteristics of chronic arthritis. We propose that oral phosphatidylcholine may be a preventive approach in ameliorating experimental rheumatoid arthritis-induced joint damage. PMID- 19296837 TI - Towards the Atlas of human African trypanosomiasis. AB - BACKGROUND: Updated, accurate and comprehensive information on the distribution of human African trypanosomiasis (HAT), also known as sleeping sickness, is critically important to plan and monitor control activities. We describe input data, methodology, preliminary results and future prospects of the HAT Atlas initiative, which will allow major improvements in the understanding of the spatial distribution of the disease. METHODS: Up-to-date as well as historical data collected by national sleeping sickness control programmes, non-governmental organizations and research institutes have been collated over many years by the HAT Control and Surveillance Programme of the World Health Organization. This body of information, unpublished for the most part, is now being screened, harmonized, and analysed by means of database management systems and geographical information systems (GIS). The number of new HAT cases and the number of people screened within a defined geographical entity were chosen as the key variables to map disease distribution in sub-Saharan Africa. RESULTS: At the time of writing, over 600 epidemiological reports and files from seventeen countries were collated and included in the data repository. The reports contain information on approximately 20,000 HAT cases, associated to over 7,000 different geographical entities. The oldest epidemiological records considered so far date back to 1985, the most recent having been gathered in 2008. Data from Cameroon, Central African Republic, Chad, Congo, Equatorial Guinea and Gabon from the year 2000 onwards were fully processed and the preliminary regional map of HAT distribution is presented. CONCLUSION: The use of GIS tools and geo-referenced, village-level epidemiological data allow the production of maps that substantially improve on the spatial quality of previous cartographic products of similar scope. The significant differences between our preliminary outputs and earlier maps of HAT transmission areas demonstrate the strong need for this systematic approach to mapping sleeping sickness and point to the inaccuracy of any calculation of population at risk based on previous maps of HAT transmission areas. The Atlas of HAT will lay the basis for novel, evidence-based methodologies to estimate the population at risk and the burden of disease, ultimately leading to more efficient targeting of interventions. Also, the Atlas will help streamline future field data collection in those parts of Africa that still require it. PMID- 19296838 TI - A giant hemolymphangioma of the pancreas in a 20-year-old girl: a report of one case and review of the literature. AB - BACKGROUND: Hemolymphangioma of the pancreas is a very rare benign tumor. There were only six reports of this disease until December 2008. Herein, we report a case of giant hemolymphangioma of the pancreas in a 20-year-old girl. CASE PRESENTATION: We describe a 20-year-old girl who presented with a mass in abdominal cavity and epigastric discomfort about a week. Physical examination showed a great abdominal mass. Abdominal computed tomography showed extrinsic duodenal compression due to a large retroperitoneal tumor possibly arising from pancreas. The tumor enucleation was performed and a diagnosis of hemolymphangioma of the pancreas was made. The patient had a complication of chylous leakage, which was successfully managed. The patient is alive and well, after 26 months of follow-up, with no complaints or recurrence. CONCLUSION: From this case and literature, we can conclude that hemolymphangioma of the pancreas in adult is a rare benign tumor, and accurate diagnosis can not be preoperatively established. Tumor resection should be performed whenever possible. The risk of recurrence seems very low. PMID- 19296839 TI - Omega 3 fatty acids reduce myeloid progenitor cell frequency in the bone marrow of mice and promote progenitor cell differentiation. AB - BACKGROUND: Omega 3 fatty acids have been found to inhibit proliferation, induce apoptosis, and promote differentiation in various cell types. The processes of cell survival, expansion, and differentiation are of key importance in the regulation of hematopoiesis. We investigated the role of omega 3 fatty acids in controlling the frequency of various myeloid progenitor cells in the bone marrow of mice. Increased progenitor cell frequency and blocked differentiation are characteristics of hematopoietic disorders of the myeloid lineage, such as myeloproliferative diseases and myeloid leukemias. RESULTS: We found that increasing the proportion of omega 3 fatty acids relative to the proportion of omega 6 fatty acids in the diet caused increased differentiation and reduced the frequency of myeloid progenitor cells in the bone marrow of mice. Furthermore, this had no adverse effect on peripheral white blood cell counts. CONCLUSION: Our results indicate that omega 3 fatty acids impact hematopoietic differentiation by reducing myeloid progenitor cell frequency in the bone marrow and promoting progenitor cell differentiation. Further exploration of this discovery could lead to the use of omega 3 fatty acids as a therapeutic option for patients that have various disorders of hematopoiesis. PMID- 19296840 TI - Discrepant comorbidity between minority and white suicides: a national multiple cause-of-death analysis. AB - BACKGROUND: Clinician training deficits and a low and declining autopsy rate adversely impact the quality of death certificates in the United States. Self report and records data for the general population indicate that proximate mental and physical health of minority suicides was at least as poor as that of white suicides. METHODS: This cross-sectional mortality study uses data from Multiple Cause-of-Death (MCOD) public use files for 1999-2003 to describe and evaluate comorbidity among black, Hispanic, and white suicides. Unintentional injury decedents are the referent for multivariate analyses. RESULTS: One or more mentions of comorbid psychopathology are documented on the death certificates of 8% of white male suicides compared to 4% and 3% of black and Hispanic counterparts, respectively. Corresponding female figures are 10%, 8%, and 6%. Racial-ethnic discrepancies in the prevalence of comorbid physical disease are more attenuated. Cross-validation with National Violent Death Reporting System data reveals high relative under-enumeration of comorbid depression/mood disorders and high relative over-enumeration of schizophrenia on the death certificates of both minorities. In all three racial-ethnic groups, suicide is positively associated with depression/mood disorders [whites: adjusted odds ratio (AOR) = 31.9, 95% CI = 29.80-34.13; blacks: AOR = 60.9, 95% CI = 42.80-86.63; Hispanics: AOR = 34.7, 95% CI = 23.36-51.62] and schizophrenia [whites: AOR = 2.4, 95% CI = 2.07-2.86; blacks: AOR = 4.2, 95% CI = 2.73-6.37; Hispanics: AOR = 4.1, 95% CI = 2.01-8.22]. Suicide is positively associated with cancer in whites [AOR = 1.8, 95% CI = 1.69-1.93] and blacks [AOR = 1.8, 95% CI = 1.36-2.48], but not with HIV or alcohol and other substance use disorders in any group under review. CONCLUSION: The multivariate analyses indicate high consistency in predicting suicide-associated comorbidities across racial-ethnic groups using MCOD data. However, low prevalence of documented comorbid psychopathology in suicides, and concomitant racial-ethnic discrepancies underscore the need for training in death certification, and routinization and standardization of timely psychological autopsies in all cases of suicide, suspected suicide, and other traumatic deaths of equivocal cause. PMID- 19296842 TI - Human articular chondrocytes express 15-lipoxygenase-1 and -2: potential role in osteoarthritis. AB - INTRODUCTION: 15-Lipoxygenases and their metabolites have been shown to exhibit anti-inflammatory and immunomodulatory properties, but little is known regarding their expression and function in chondrocytes. The objective of this study was to evaluate the expression of 15-lipoxygenase-1 and -2 in human articular chondrocytes, and to investigate the effects of their metabolites 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids on IL-1beta-induced matrix metalloproteinase (MMP)-1 and MMP-13 expression. METHODS: The expression levels of 15-lipoxygenase-1 and -2 were analyzed by reverse transcription PCR and Western blotting in chondrocytes, and by immunohistochemistry in cartilage. Chondrocytes or cartilage explants were stimulated with IL-1beta in the absence or presence of 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids, and the levels of MMP-1 and MMP-13 protein production and type II collagen cleavage were evaluated using immunoassays. The role of peroxisome proliferator activated receptor (PPAR)gamma was evaluated using transient transfection experiments and the PPARgamma antagonist GW9662. RESULTS: Articular chondrocytes express 15-lipoxygenase-1 and -2 at the mRNA and protein levels. 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids dose dependently decreased IL-1beta-induced MMP-1 and MMP-13 protein and mRNA expression as well as type II collagen cleavage. The effect on MMP-1 and MMP-13 expression does not require de novo protein synthesis. 13(S)-hydroxy octadecadienoic and 15(S) hydroxyeicosatetraenoic acids activated endogenous PPARgamma, and GW9662 prevented their suppressive effect on MMP-1 and MMP-13 production, suggesting the involvement of PPARgamma in these effects. CONCLUSIONS: This study is the first to demonstrate the expression of 15-lipoxygenase-1 and -2 in articular chondrocytes. Their respective metabolites, namely 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids, suppressed IL-1beta-induced MMP-1 and MMP-13 expression in a PPARgamma-dependent pathway. These data suggest that 15 lipoxygenases may have chondroprotective properties by reducing MMP-1 and MMP-13 expression. PMID- 19296843 TI - Primary care physicians' reported use of pre-screening discussions for prostate cancer screening: a cross-sectional survey. AB - BACKGROUND: Professional medical organizations recommend individualized patient decision making about prostate cancer screening. Little is known about primary care physicians' use of pre-screening discussions to promote informed decision making for prostate cancer screening. The aim of this study is to explore physicians' use of pre-screening discussions and reasons why physicians would or would not try to persuade patients to be screened if they initially refuse testing. METHODS: Primary care physicians completed a self-administered survey about prostate cancer screening practices for informed decision making. RESULTS: Sixty-six physicians (75.9%) completed the survey, and 63 were used in the analysis. Thirteen physicians (20.6%) reported not using prescreening discussions, 45 (71.4%) reported the use of prescreening discussions, and 3 (4.8%) reported neither ordering the PSA test nor discussing it with patients. Sixty-nine percent of physicians who reported not having discussions indicated they were more likely to screen African American patients for prostate cancer, compared to 50% of physicians who reported the use of discussions (Chi-square(1) = 1.62, p = .20). Similarly, 91% of physicians who reported not having discussions indicated they are more likely to screen patients with a family history of prostate cancer, compared to 46% of those who reported the use of discussion (Chi-square(1) = 13.27, p < .001). Beliefs about the scientific evidence and efficacy of screening, ethical concerns regarding patient autonomy, and concerns about time constraints differed between physicians who would and would not try to persuade a patient to be tested. CONCLUSION: Although guidelines recommend discussing the risks and benefits of prostate cancer screening, physicians report varying practice styles. Future research needs to consider the nature of discussions and the degree to which informed decision making is being achieved in clinical practice. PMID- 19296844 TI - Towards agreement on best practice for publishing raw clinical trial data. AB - Many research-funding agencies now require open access to the results of research they have funded, and some also require that researchers make available the raw data generated from that research. Similarly, the journal Trials aims to address inadequate reporting in randomised controlled trials, and in order to fulfil this objective, the journal is working with the scientific and publishing communities to try to establish best practice for publishing raw data from clinical trials in peer-reviewed biomedical journals. Common issues encountered when considering raw data for publication include patient privacy - unless explicit consent for publication is obtained - and ownership, but agreed-upon policies for tackling these concerns do not appear to be addressed in the guidance or mandates currently established. Potential next steps for journal editors and publishers, ethics committees, research-funding agencies, and researchers are proposed, and alternatives to journal publication, such as restricted access repositories, are outlined. PMID- 19296845 TI - Effect of the consumption of a fermented dairy product containing Bifidobacterium lactis DN-173 010 on constipation in childhood: a multicentre randomised controlled trial (NTRTC: 1571). AB - BACKGROUND: Constipation is a frustrating symptom affecting 3% of children worldwide. Randomised controlled trials show that both polyethylene glycol and lactulose are effective in increasing defecation frequency in children with constipation. However, in 30-50%, these children reported abdominal pain, bloating, flatulence, diarrhoea, nausea and bad taste of the medication. Two recent studies have shown that the fermented dairy product containing Bifidobacterium lactis strain DN-173 010 is effective in increasing stool frequency in constipation-predominant irritable bowel syndrome patients with a defecation frequency < 3/week and in constipated women with a defecation frequency < 3/week. Goal of this study is to determine whether this fermented dairy product is effective in the treatment of constipated children with a defecation frequency < 3/week. METHODS/DESIGN: It is a two nation (The Netherlands and Poland) double-blind, placebo-controlled randomised multicentre trial in which 160 constipated children (age 3-16 years) with a defecation frequency <3/week will be randomly allocated to consume a fermented dairy product containing Bifidobacterium lactis DN-173 010 or a control product, twice a day, for 3 weeks. During the study all children are instructed to try to defecate on the toilet for 5-10 minutes after each meal (3 times a day) and daily complete a standardized bowel diary. Primary endpoint is stool frequency. Secondary endpoints are stool consistency, faecal incontinence frequency, pain during defecation, digestive symptoms (abdominal pain, flatulence), adverse effects (nausea, diarrhoea, bad taste) and intake of rescue medication (Bisacodyl). Rate of success and rate of responders are also evaluated, with success defined as > or = 3 bowel movements per week and < or =1 faecal incontinence episode over the last 2 weeks of product consumption and responder defined as a subject reporting a stool frequency > or = 3 on the last week of product consumption. To demonstrate that the success percentage in the intervention group will be 35% and the success percentage in the control group (acidified milk without ferments, toilet training, bowel diary) will be 15%, with alpha 0.05 and power 80%, a total sample size of 160 patients was calculated. CONCLUSION: This study is aimed to show that the fermented dairy product containing Bifidobacterium lactis strain DN 173 010 is effective in increasing stool frequency after 3 weeks of product consumption in children with functional constipation and a defecation frequency < 3/week. PMID- 19296846 TI - Factors related to treatment intensity in Swiss primary care. AB - BACKGROUND: Questions about the existence of supplier-induced demand emerge repeatedly in discussions about governing Swiss health care. This study therefore aimed to evaluate the interrelationship between structural factors of supply and the volume of services that are provided by primary care physicians in Switzerland. METHODS: The study was designed as a cross-sectional investigation, based on the complete claims data from all Swiss health care insurers for the year 2004, which covered information from 6087 primary care physicians and 4.7 million patients. Utilization-based health service areas were constructed and used as spatial units to analyze effects of density of supply. Hierarchical linear models were applied to analyze the data. RESULTS: The data showed that, within a service area, a higher density of primary care physicians was associated with higher mortality rates and specialist density but not with treatment intensity in primary care. Higher specialist density was weakly associated with higher mortality rates and with higher treatment intensity density of primary care physicians. Annual physician-level data indicate a disproportionate increase of supplied services irrespective of the size of the number of patients treated during the same year and, even in high volume practices, no rationing but a paradoxical inducement of consultations occurred. The results provide empirical evidence that higher densities of primary care physicians, specialists and the availability of out-patient hospital clinics in a given area are associated with higher volume of supplied services per patient in primary care practices. Analyses stratified by language regions showed differences that emphasize the effect of the cantonal based (fragmented) governance of Swiss health care. CONCLUSION: The study shows high volumes in Swiss primary care and provides evidence that the volume of supply is not driven by medical needs alone. Effects related to the competition for patients between primary care physicians, specialists and out-patient hospital clinics and an association with the system of reimbursing services on a fee-for-service basis can not be excluded. PMID- 19296847 TI - Structural pattern matching of nonribosomal peptides. AB - BACKGROUND: Nonribosomal peptides (NRPs), bioactive secondary metabolites produced by many microorganisms, show a broad range of important biological activities (e.g. antibiotics, immunosuppressants, antitumor agents). NRPs are mainly composed of amino acids but their primary structure is not always linear and can contain cycles or branchings. Furthermore, there are several hundred different monomers that can be incorporated into NRPs. The NORINE database, the first resource entirely dedicated to NRPs, currently stores more than 700 NRPs annotated with their monomeric peptide structure encoded by undirected labeled graphs. This opens a way to a systematic analysis of structural patterns occurring in NRPs. Such studies can investigate the functional role of some monomeric chains, or analyse NRPs that have been computationally predicted from the synthetase protein sequence. A basic operation in such analyses is the search for a given structural pattern in the database. RESULTS: We developed an efficient method that allows for a quick search for a structural pattern in the NORINE database. The method identifies all peptides containing a pattern substructure of a given size. This amounts to solving a variant of the maximum common subgraph problem on pattern and peptide graphs, which is done by computing cliques in an appropriate compatibility graph. CONCLUSION: The method has been incorporated into the NORINE database, available at http://bioinfo.lifl.fr/norine. Less than one second is needed to search for a pattern in the entire database. PMID- 19296848 TI - Dysregulated apoptosis and NFkappaB expression in COPD subjects. AB - BACKGROUND: The abnormal regulation of neutrophil apoptosis may contribute to the ineffective resolution of inflammation in chronic lung diseases. Multiple signalling pathways are implicated in regulating granulocyte apoptosis, in particular, NFkappaB (nuclear factor-kappa B) signalling which delays constitutive neutrophil apoptosis. Although some studies have suggested a dysregulation in the apoptosis of airway cells in chronic obstructive pulmonary disease (COPD), no studies to date have directly investigated if NFkappaB is associated with apoptosis of airway neutrophils from COPD patients. The objectives of this study were to examine spontaneous neutrophil apoptosis in stable COPD subjects (n = 13), healthy smoking controls (n = 9) and non-smoking controls (n = 9) and to investigate whether the neutrophil apoptotic process in inflammatory conditions is associated with NFkappaB activation. METHODS: Analysis of apoptosis in induced sputum was carried out by 3 methods; light microscopy, Annexin V/Propidium iodide and the terminal transferase-mediated dUTP nick end labeling (TUNEL) method. Activation of NFkappaB was assessed using a flow cytometric method and the phosphorylation state of IkappaBalpha was carried out using the Bio-Rad Bio-Plex phosphoprotein IkappaBalpha assay. RESULTS: Flow cytometric analysis showed a significant reduction in the percentage of sputum neutrophils undergoing spontaneous apoptosis in healthy smokers and subjects with COPD compared to non-smokers (p < 0.001). Similar findings were demonstrated using the Tunel assay and in the morphological identification of apoptotic neutrophils. A significant increase was observed in the expression of both the p50 (p = 0.006) and p65 (p = 0.006) subunits of NFkappaB in neutrophils from COPD subjects compared to non-smokers. CONCLUSION: These results demonstrate that apoptosis is reduced in the sputum of COPD subjects and in healthy control smokers and may be regulated by an associated activation of NFkappaB. PMID- 19296849 TI - A compatible interaction of Alternaria brassicicola with Arabidopsis thaliana ecotype DiG: evidence for a specific transcriptional signature. AB - BACKGROUND: The interaction of Arabidopsis with Alternaria brassicicola provides a model for disease caused by necrotrophs, but a drawback has been the lack of a compatible pathosystem. Infection of most ecotypes, including the widely-studied line Col-0, with this pathogen generally leads to a lesion that does not expand beyond the inoculated area. This study examines an ecotype, Dijon G (DiG), which is considered sensitive to A. brassicicola. RESULTS: We show that the interaction has the characteristics of a compatible one, with expanding rather than limited lesions. To ask whether DiG is merely more sensitive to the pathogen or, rather, interacts in distinct manner, we identified genes whose regulation differs between Col-0 and DiG challenged with A. brassicicola. Suppression subtractive hybridization was used to identify differentially expressed genes, and their expression was verified using semi-quantitative PCR. We also tested a set of known defense-related genes for differential regulation in the two plant-pathogen interactions. Several known pathogenesis-related (PR) genes are up-regulated in both interactions. PR1, and a monooxygenase gene identified in this study, MO1, are preferentially up-regulated in the compatible interaction. In contrast, GLIP1, which encodes a secreted lipase, and DIOX1, a pathogen-response related dioxygenase, are preferentially up-regulated in the incompatible interaction. CONCLUSION: The results show that DiG is not only more susceptible, but demonstrate that its interaction with A. brassicicola has a specific transcriptional signature. PMID- 19296851 TI - Reducing dimensionality for prediction of genome-wide breeding values. AB - Partial least square regression (PLSR) and principal component regression (PCR) are methods designed for situations where the number of predictors is larger than the number of records. The aim was to compare the accuracy of genome-wide breeding values (EBV) produced using PLSR and PCR with a Bayesian method, 'BayesB'. Marker densities of 1, 2, 4 and 8 Ne markers/Morgan were evaluated when the effective population size (Ne) was 100. The correlation between true breeding value and estimated breeding value increased with density from 0.611 to 0.681 and 0.604 to 0.658 using PLSR and PCR respectively, with an overall advantage to PLSR of 0.016 (s.e = 0.008). Both methods gave a lower accuracy compared to the 'BayesB', for which accuracy increased from 0.690 to 0.860. PLSR and PCR appeared less responsive to increased marker density with the advantage of 'BayesB' increasing by 17% from a marker density of 1 to 8Ne/M. PCR and PLSR showed greater bias than 'BayesB' in predicting breeding values at all densities. Although, the PLSR and PCR were computationally faster and simpler, these advantages do not outweigh the reduction in accuracy, and there is a benefit in obtaining relevant prior information from the distribution of gene effects. PMID- 19296850 TI - Genetic identity and differential gene expression between Trichomonas vaginalis and Trichomonas tenax. AB - BACKGROUND: Trichomonas vaginalis is a human urogenital pathogen responsible for trichomonosis, the number-one, non-viral sexually transmitted disease (STD) worldwide, while T. tenax is a commensal of the human oral cavity, found particularly in patients with poor oral hygiene and advanced periodontal disease. The extent of genetic identity between T. vaginalis and its oral commensal counterpart is unknown. RESULTS: Genes that were differentially expressed in T. vaginalis were identified by screening three independent subtraction cDNA libraries enriched for T. vaginalis genes. The same thirty randomly selected cDNA clones encoding for proteins with specific functions associated with colonization were identified from each of the subtraction cDNA libraries. In addition, a T. vaginalis cDNA expression library was screened with patient sera that was first pre-adsorbed with an extract of T. tenax antigens, and seven specific cDNA clones were identified from this cDNA library. Interestingly, some of the clones identified by the subtraction cDNA screening were also obtained from the cDNA expression library with the pre-adsorbed sera. Moreover and noteworthy, clones identified by both the procedures were found to be up-regulated in expression in T. vaginalis upon contact with vaginal epithelial cells, suggesting a role for these gene products in host colonization. Semi-quantitative RT-PCR analysis of select clones showed that the genes were not unique to T. vaginalis and that these genes were also present in T. tenax, albeit at very low levels of expression. CONCLUSION: These results suggest that T. vaginalis and T. tenax have remarkable genetic identity and that T. vaginalis has higher levels of gene expression when compared to that of T. tenax. The data may suggest that T. tenax could be a variant of T. vaginalis. PMID- 19296852 TI - Research and development of a new RF-assisted device for bloodless rapid transection of the liver: computational modeling and in vivo experiments. AB - BACKGROUND: Efficient and safe transection of biological tissue in liver surgery is strongly dependent on the ability to address both parenchymal division and hemostasis simultaneously. In addition to the conventional clamp crushing or finger fracture methods other techniques based on radiofrequency (RF) currents have been extensively employed to reduce intraoperative blood loss. In this paper we present our broad research plan for a new RF-assisted device for bloodless, rapid resection of the liver. METHODS: Our research plan includes computer modeling and in vivo studies. Computer modeling was based on the Finite Element Method (FEM) and allowed us to estimate the distribution of electrical power deposited in the tissue, along with assessing the effect of the characteristics of the device on the temperature profiles. Studies based on in vivo pig liver models provided a comparison of the performance of the new device with other techniques (saline-linked technology) currently employed in clinical practice. Finally, the plan includes a pilot clinical trial, in which both the new device and the accessory equipment are seen to comply with all safety requirements. RESULTS: The FEM results showed a high electrical gradient around the tip of the blade, responsible for the maximal increase of temperature at that point, where temperature reached 100 degrees C in only 3.85 s. Other hot points with lower temperatures were located at the proximal edge of the device. Additional simulations with an electrically insulated blade produced more uniform and larger lesions (assessed as the 55 degrees C isotherm) than the electrically conducting blade. The in vivo study, in turn, showed greater transection speed (3 +/- 0 and 3 +/- 1 cm2/min for the new device in the open and laparoscopic approaches respectively) and also lower blood loss (70 +/- 74 and 26 +/- 34 mL) during transection of the liver, as compared to saline-linked technology (2 +/- 1 cm2/min with P = 0.002, and 527 +/- 273 mL with P = 0.001). CONCLUSION: A new RF assisted device for bloodless, rapid liver resection was designed, built and tested. The results demonstrate the potential advantages of this device over others currently employed. PMID- 19296854 TI - Molecular determinants archetypical to the phylum Nematoda. AB - BACKGROUND: Nematoda diverged from other animals between 600-1,200 million years ago and has become one of the most diverse animal phyla on earth. Most nematodes are free-living animals, but many are parasites of plants and animals including humans, posing major ecological and economical challenges around the world. RESULTS: We investigated phylum-specific molecular characteristics in Nematoda by exploring over 214,000 polypeptides from 32 nematode species including 27 parasites. Over 50,000 nematode protein families were identified based on primary sequence, including approximately 10% with members from at least three different species. Nearly 1,600 of the multi-species families did not share homology to Pfam domains, including a total of 758 restricted to Nematoda. Majority of the 462 families that were conserved among both free-living and parasitic species contained members from multiple nematode clades, yet approximately 90% of the 296 parasite-specific families originated only from a single clade. Features of these protein families were revealed through extrapolation of essential functions from observed RNAi phenotypes in C. elegans, bioinformatics-based functional annotations, identification of distant homology based on protein folds, and prediction of expression at accessible nematode surfaces. In addition, we identified a group of nematode-restricted sequence features in energy-generating electron transfer complexes as potential targets for new chemicals with minimal or no toxicity to the host. CONCLUSION: This study identified and characterized the molecular determinants that help in defining the phylum Nematoda, and therefore improved our understanding of nematode protein evolution and provided novel insights for the development of next generation parasite control strategies. PMID- 19296853 TI - A computational approach for genome-wide mapping of splicing factor binding sites. AB - Alternative splicing is regulated by splicing factors that serve as positive or negative effectors, interacting with regulatory elements along exons and introns. Here we present a novel computational method for genome-wide mapping of splicing factor binding sites that considers both the genomic environment and the evolutionary conservation of the regulatory elements. The method was applied to study the regulation of different alternative splicing events, uncovering an interesting network of interactions among splicing factors. PMID- 19296855 TI - Rational mutagenesis to support structure-based drug design: MAPKAP kinase 2 as a case study. AB - BACKGROUND: Structure-based drug design (SBDD) can provide valuable guidance to drug discovery programs. Robust construct design and expression, protein purification and characterization, protein crystallization, and high-resolution diffraction are all needed for rapid, iterative inhibitor design. We describe here robust methods to support SBDD on an oral anti-cytokine drug target, human MAPKAP kinase 2 (MK2). Our goal was to obtain useful diffraction data with a large number of chemically diverse lead compounds. Although MK2 structures and structural methods have been reported previously, reproducibility was low and improved methods were needed. RESULTS: Our construct design strategy had four tactics: N- and C-terminal variations; entropy-reducing surface mutations; activation loop deletions; and pseudoactivation mutations. Generic, high throughput methods for cloning and expression were coupled with automated liquid dispensing for the rapid testing of crystallization conditions with minimal sample requirements. Initial results led to development of a novel, customized robotic crystallization screen that yielded MK2/inhibitor complex crystals under many conditions in seven crystal forms. In all, 44 MK2 constructs were generated, ~500 crystals were tested for diffraction, and ~30 structures were determined, delivering high-impact structural data to support our MK2 drug design effort. CONCLUSION: Key lessons included setting reasonable criteria for construct performance and prioritization, a willingness to design and use customized crystallization screens, and, crucially, initiation of high-throughput construct exploration very early in the drug discovery process. PMID- 19296856 TI - Evolution of DNA polymerases: an inactivated polymerase-exonuclease module in Pol epsilon and a chimeric origin of eukaryotic polymerases from two classes of archaeal ancestors. AB - BACKGROUND: Evolution of DNA polymerases, the key enzymes of DNA replication and repair, is central to any reconstruction of the history of cellular life. However, the details of the evolutionary relationships between DNA polymerases of archaea and eukaryotes remain unresolved. RESULTS: We performed a comparative analysis of archaeal, eukaryotic, and bacterial B-family DNA polymerases, which are the main replicative polymerases in archaea and eukaryotes, combined with an analysis of domain architectures. Surprisingly, we found that eukaryotic Polymerase epsilon consists of two tandem exonuclease-polymerase modules, the active N-terminal module and a C-terminal module in which both enzymatic domains are inactivated. The two modules are only distantly related to each other, an observation that suggests the possibility that Pol epsilon evolved as a result of insertion and subsequent inactivation of a distinct polymerase, possibly, of bacterial descent, upstream of the C-terminal Zn-fingers, rather than by tandem duplication. The presence of an inactivated exonuclease-polymerase module in Pol epsilon parallels a similar inactivation of both enzymatic domains in a distinct family of archaeal B-family polymerases. The results of phylogenetic analysis indicate that eukaryotic B-family polymerases, most likely, originate from two distantly related archaeal B-family polymerases, one form giving rise to Pol epsilon, and the other one to the common ancestor of Pol alpha, Pol delta, and Pol zeta. The C-terminal Zn-fingers that are present in all eukaryotic B-family polymerases, unexpectedly, are homologous to the Zn-finger of archaeal D-family DNA polymerases that are otherwise unrelated to the B family. The Zn-finger of Polepsilon shows a markedly greater similarity to the counterpart in archaeal PolD than the Zn-fingers of other eukaryotic B-family polymerases. CONCLUSION: Evolution of eukaryotic DNA polymerases seems to have involved previously unnoticed complex events. We hypothesize that the archaeal ancestor of eukaryotes encoded three DNA polymerases, namely, two distinct B-family polymerases and a D family polymerase all of which contributed to the evolution of the eukaryotic replication machinery. The Zn-finger might have been acquired from PolD by the B family form that gave rise to Pol epsilon prior to or in the course of eukaryogenesis, and subsequently, was captured by the ancestor of the other B family eukaryotic polymerases. The inactivated polymerase-exonuclease module of Pol epsilon might have evolved by fusion with a distinct polymerase, rather than by duplication of the active module of Pol epsilon, and is likely to play an important role in the assembly of eukaryotic replication and repair complexes. PMID- 19296857 TI - The PHA Depolymerase Engineering Database: A systematic analysis tool for the diverse family of polyhydroxyalkanoate (PHA) depolymerases. AB - BACKGROUND: Polyhydroxyalkanoates (PHAs) can be degraded by many microorganisms using intra- or extracellular PHA depolymerases. PHA depolymerases are very diverse in sequence and substrate specificity, but share a common alpha/beta hydrolase fold and a catalytic triad, which is also found in other alpha/beta hydrolases. RESULTS: The PHA Depolymerase Engineering Database (DED, http://www.ded.uni-stuttgart.de) has been established as a tool for systematic analysis of this enzyme family. The DED contains sequence entries of 587 PHA depolymerases, which were assigned to 8 superfamilies and 38 homologous families based on their sequence similarity. For each family, multiple sequence alignments and profile hidden Markov models are provided, and functionally relevant residues are annotated. CONCLUSION: The DED is a valuable tool which can be applied to identify new PHA depolymerase sequences from complete genomes in silico, to classify PHA depolymerases, to predict their biochemical properties, and to design enzyme variants with improved properties. PMID- 19296858 TI - Bayesian clustering and feature selection for cancer tissue samples. AB - BACKGROUND: The versatility of DNA copy number amplifications for profiling and categorization of various tissue samples has been widely acknowledged in the biomedical literature. For instance, this type of measurement techniques provides possibilities for exploring sets of cancerous tissues to identify novel subtypes. The previously utilized statistical approaches to various kinds of analyses include traditional algorithmic techniques for clustering and dimension reduction, such as independent and principal component analyses, hierarchical clustering, as well as model-based clustering using maximum likelihood estimation for latent class models. RESULTS: While purely algorithmic methods are usually easily applicable, their suboptimal performance and limitations in making formal inference have been thoroughly discussed in the statistical literature. Here we introduce a Bayesian model-based approach to simultaneous identification of underlying tissue groups and the informative amplifications. The model-based approach provides the possibility of using formal inference to determine the number of groups from the data, in contrast to the ad hoc methods often exploited for similar purposes. The model also automatically recognizes the chromosomal areas that are relevant for the clustering. CONCLUSION: Validatory analyses of simulated data and a large database of DNA copy number amplifications in human neoplasms are used to illustrate the potential of our approach. Our software implementation BASTA for performing Bayesian statistical tissue profiling is freely available for academic purposes at (http://web.abo.fi/fak/mnf/mate/jc/software/basta.html). PMID- 19296859 TI - Reducing the bias of estimates of genotype by environment interactions in random regression sire models. AB - The combination of a sire model and a random regression term describing genotype by environment interactions may lead to biased estimates of genetic variance components because of heterogeneous residual variance. In order to test different models, simulated data with genotype by environment interactions, and dairy cattle data assumed to contain such interactions, were analyzed. Two animal models were compared to four sire models. Models differed in their ability to handle heterogeneous variance from different sources. Including an individual effect with a (co)variance matrix restricted to three times the sire (co)variance matrix permitted the modeling of the additive genetic variance not covered by the sire effect. This made the ability of sire models to handle heterogeneous genetic variance approximately equivalent to that of animal models. When residual variance was heterogeneous, a different approach to account for the heterogeneity of variance was needed, for example when using dairy cattle data in order to prevent overestimation of genetic heterogeneity of variance. Including environmental classes can be used to account for heterogeneous residual variance. PMID- 19296860 TI - Multitasking human endometrium: a review of endometrial biopsy as a diagnostic tool, therapeutic applications, and a source of adult stem cells. AB - The endometrium is one of the most fascinating tissues in the human body. Its sole purpose is to enable implantation of an embryo during a very short window of opportunity in the menstrual cycle. When implantation does not take place, the endometrial cells are shed, enabling growth of a new endometrial layer the following month. Recently recognized stem cells may be the source through which this physiological turnover can take place. These cells, however, may also result in pathological conditions such as endometriosis, adenomyosis, and carcinoma when control by surrounding niche cells is lost. The methods to obtain endometrial tissue are critically reviewed, including the various catheters compared with the Pipelle. Recent advances in molecular studies of the endometrium enable researchers to define markers of implantation, and study the interaction between the embryo and the endometrium in vitro. Moreover, mechanisms by which wounding the endometrium by the biopsy catheter actually seems to improve embryo implantation are hypothesized. Finally, detection, isolation, and differentiation of adult endometrial stem cells have the potential to provide a resource for therapy of chronic conditions. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians Leaning Objectives: After completion of this article, the reader should be able to explain the concept of the endometrium as a multi tasking tissue, describe the role in health and potential role in medical care of adult stem cells in the endometrium, and describe the clinical characteristics of endometrial biopsy techniques. PMID- 19296862 TI - The area centralis in the chicken retina contains efferent target amacrine cells. AB - The retinas of birds receive a substantial efferent, or centrifugal, input from a midbrain nucleus. The function of this input is presently unclear, but previous work in the pigeon has shown that efferent input is excluded from the area centralis, suggesting that the functions of the area centralis and the efferent system are incompatible. Using an antibody specific to rods, we have identified the area centralis in another species, the chicken, and mapped the distribution of the unique amacrine cells that are the postsynaptic partners of efferent fibers. Efferent target amacrine cells are found within the chicken area centralis and their density is continuous across the border of the area centralis. In contrast to the pigeon retina then, we conclude that the chicken area centralis receives efferent input. We suggest that the difference between the two species is attributable to the presence of a fovea within the area centralis of the pigeon and its absence from that of the chicken. PMID- 19296863 TI - Efficiency of Stenger test in confirming profound, unilateral pseudohypacusis. AB - OBJECTIVE: Conscious and deceptive exaggeration of hearing loss is termed pseudohypacusis. Even though the Stenger test has been used in the management of pseudohypacusis for almost a century, its sensitivity, specificity and predictive values for unilateral pseudohypacusis have not previously been reported, to our best knowledge. We investigated the efficiency of the Stenger test in detecting unilateral pseudohypacusis, accepting auditory brainstem response testing as the 'gold standard'. MATERIALS AND METHODS: Candidates with questionable profound or total hearing loss were enrolled in the study. Pure tone audiometry, speech and tonal Stenger tests, and click test auditory brainstem response measurement were performed. Accepting auditory brainstem response testing as the gold standard, the sensitivity, specificity and predictive values of the Stenger test for unilateral, profound pseudohypacusis were assessed. RESULTS: Two hundred military candidates were enrolled in the study. The sensitivity and specificity of the Stenger test in verifying unilateral, profound hearing loss were 99.4 and 70 per cent, respectively. The positive and negative predictive values of the test were 87.5 and 98.4 per cent, respectively. CONCLUSION: The Stenger test is widely used for the evaluation of unilateral or asymmetrical pseudohypacusis. In our opinion, it is a powerfully reliable test. More difficult cases require objective electrophysiological testing to verify functional hearing loss and to exclude specific diagnoses that may imitate pseudohypacusis. PMID- 19296861 TI - How disturbed sleep may be a risk factor for adverse pregnancy outcomes. AB - Adverse pregnancy outcomes associated with significant maternal and infant morbidity are on the rise in Western society despite advances of medical technology. Current risk factors are insufficient to identify women at greatest risk of developing an adverse outcome. An attempt to identify novel contributors to increased risk is warranted. Sleep disturbances are frequent during pregnancy, yet are often dismissed as irrelevant. Emerging evidence indicates that sleep disturbances are associated with poor health outcomes, including cardiovascular disease. Disturbed sleep is also linked with an increased inflammatory response. Increased inflammation is proposed as a key biological pathway through which chronic disease and adverse pregnancy outcomes develop. In this paper, we propose a model and a testable hypothesis of how disturbed sleep in the first 20 weeks of pregnancy could contribute to adverse pregnancy outcomes such as preeclampsia, intrauterine growth restriction, and preterm birth via increased inflammation. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians Leaning Objectives: After completion of this article, the reader should be able to outline data linking sleep disturbances with an increased risk of some systemic disorders, recall characteristics of pregnancy complications which support the hypothesis that sleep disturbances may be related to these pregnancy outcomes, and summarize the likelihood and types of sleep disturbances that are common in pregnant women. PMID- 19296864 TI - Trans-nasal injection of botulinum toxin. AB - INTRODUCTION: Office-based laryngeal injection for the treatment of a variety of voice disorders is an established practice. Various methods of delivery to the vocal folds are in practice. AIM: We present a simple and repeatable method of injecting botulinum toxin into the larynx. METHOD: Botulinum toxin A (Botox) was delivered into the larynx using a channelled fibre-optic laryngoscope under local anaesthetic, in an office setting. RESULTS: Seven patients received regular botulinum toxin injections, via our preferred method. The treatment intervals and dosage regime varied depending on patient requirements. The procedure was carried out in an office setting, with minimal discomfort and inconvenience to the patient. CONCLUSION: We present a method of botulinum toxin delivery to the larynx which is simple, easily repeatable and simply learnt by the otolaryngologist familiar with fibre-optic laryngoscope usage. This method is also comfortable and convenient for the patient. PMID- 19296866 TI - Protein kinase inhibitors: contributions from structure to clinical compounds. AB - Protein kinases catalyse key phosphorylation reactions in signalling cascades that affect every aspect of cell growth, differentiation and metabolism. The kinases have become prime targets for drug intervention in the diseased state, especially in cancer. There are currently 10 drugs that have been approved for clinical use and many more in clinical trials. This review summarises the structural basis for protein kinase inhibition and discusses the mode of action for each of the approved drugs in the light of structural results. All but one of the approved compounds target the ATP binding site on the kinase. Both the active and inactive conformations of protein kinases have been used in strategies to produce potent and selective compounds. Targeting the inactive conformation can give high specificity. Targeting the active conformation is favourable where the diseased state has arisen from activating mutations, but such inhibitors generally target several protein kinases. Drug resistance mutations are a potential risk for both conformational states, where drug-binding regions are not directly involved in catalysis. Imatinib (Glivec), the most successful of protein kinase inhibitors, targets the inactive conformation of ABL tyrosine kinase. Newer compounds, such as dasatinib, which targets the ABL active state, have been developed to increase potency and have proved effective for some, but not all, drug-resistant mutations. The first epidermal growth factor receptor (EGFR) inhibitors in clinical use [gefitinib (Iressa) and erlotinib (Tarceva)] targeted the active form of the kinase, and this proved advantageous for patients whose cancer was caused by mutations that resulted in a constitutively active EGFR kinase domain. Newer approved compounds, such as lapatinib (Tykerb), target the inactive conformation with high potency. A further compound that forms a covalent attachment to the kinase has been found to overcome one of the major drug resistance mutations, where the effectiveness of the drug in vivo is dependent on its ability to compete successfully in the presence of cellular concentrations of ATP. Inhibitors of vascular endothelial growth factor receptor (VEGFR) kinase against cancer angiogenesis show the advantage of some relaxation in specificity. Sorafenib, originally developed as RAF inhibitor, is now in clinical use as a VEGFR inhibitor. Temsirolimus (a derivative of rapamycin) is the only example of a drug in clinical use that does not target the kinase ATP site. Instead rapamycin, when in complex with the protein FKBP12, effectively targets mTOR kinase at a site located on a domain, the FRB domain, that appears to be involved in localisation or substrate docking. PMID- 19296865 TI - Does major depressive disorder change with age? AB - BACKGROUND: The authors used results from a 20-year, high-intensity follow-up to measure the influence of ageing, and of age at onset, on the long-term persistence of symptoms in major depressive disorder (MDD). METHOD: Subjects who completed a 20-year series of semi-annual and then annual assessments with a stable diagnosis of MDD or schizo-affective disorder other than mainly schizophrenic (n=220) were divided according to their ages at intake into youngest (18-29 years), middle (30-44 years) and oldest (>45 years) groups. Depressive morbidity was quantified as the proportion of weeks spent in major depressive or schizo-affective episodes. General linear models then tested for effects of time and time x group interactions on these measures. Regression analyses compared the influence of age of onset and of current age. RESULTS: Analyses revealed no significant time or group x time effects on the proportions of weeks in major depressive episodes in any of the three age groups. Earlier ages of onset were associated with greater symptom persistence, particularly in the youngest group. The proportions of weeks ill showed intra-individual stability over time that was most evident in the oldest group. CONCLUSIONS: These results indicate that the persistence of depressive symptoms in MDD does not change as individuals move from their third to their fifth decade, from their fourth to their sixth decade, or from their sixth to their eighth decade. An early age of onset, rather than youth per se, is associated with greater morbidity over two decades. PMID- 19296868 TI - Effect of environmental factors on the spatio-temporal patterns of influenza spread. AB - Although spatio-temporal patterns of influenza spread often suggest that environmental factors play a role, their effect on the geographical variation in the timing of annual epidemics has not been assessed. We examined the effect of solar radiation, dew point, temperature and geographical position on the city specific timing of epidemics in the USA. Using paediatric in-patient data from hospitals in 35 cities for each influenza season in the study period 2000-2005, we determined 'epidemic timing' by identifying the week of peak influenza activity. For each city we calculated averages of daily climate measurements for 1 October to 31 December. Bayesian hierarchical models were used to assess the strength of association between each variable and epidemic timing. Of the climate variables only solar radiation was significantly related to epidemic timing (95% CI -0.027 to -0.0032). Future studies may elucidate biological mechanisms intrinsically linked to solar radiation that contribute to epidemic timing in temperate regions. PMID- 19296867 TI - A prolonged outbreak of Salmonella Typhimurium infection related to an uncommon vehicle: hard cheese made from raw milk. AB - In 2006, in The Netherlands, an outbreak of Salmonella Typhimurium phage type 561 (STM DT7, corresponding to the rare DT7 in the international typing scheme) was detected, accumulating to over 200 cases. By telephone interviews, data were collected from all laboratory-confirmed cases. In addition, in August 2006, a case-control study was performed in a subset of cases. Environmental and microbiological investigation was performed on a suspected dairy farm. In the case-control study (51 cases, 105 matched controls), hard cheese purchased from a farm, specifically farm X, and from a market stall were found to be associated with infection. The dairy production room of farm X tested STM DT7-positive in August. However, it was only in November, after earlier unsuccessful attempts, that a low-level contamination was confirmed in the hard farmhouse cheese, triggering control measures. A timely and adequate response was hampered during this outbreak for several reasons. Measures for improvement in handling future similar incidents are discussed. PMID- 19296869 TI - Risk of transmitting meningococcal infection by transient contact on aircraft and other transport. AB - Contact tracing of persons with meningococcal disease who have travelled on aeroplane or other multi-passenger transport is not consistent between countries. We searched the literature for clusters of meningococcal disease linked by transient contact on the same plane, train, bus or boat. We found reports of two clusters in children on the same school bus and one in passengers on the same plane. Cases within each of these three clusters were due to strains that were genetically indistinguishable. In the aeroplane cluster the only link between the two cases was through a single travel episode. The onset of illness (2 and 5 days after the flight) is consistent with infection from an unidentified carrier around the time of air travel. In contrast to the established risk of transmission from a case of tuberculosis, it is likely that the risk from a case of meningococcal disease to someone who is not identified as a close contact is exceedingly low. This should be considered in making international recommendations for passenger contact tracing after a case of meningococcal disease on a plane or other multi-passenger transport. PMID- 19296870 TI - A randomized controlled trial of vitamin D3 supplementation for the prevention of symptomatic upper respiratory tract infections. AB - Vitamin D has been shown to be an important immune system regulator. Vitamin D insufficiency during winter may cause increased susceptibility to upper respiratory tract infections (URIs). To determine whether vitamin D supplementation during the winter season prevents or decreases URI symptoms, 162 adults were randomized to receive 50 microg vitamin D3 (2000 IU) daily or matching placebo for 12 weeks. A bi-weekly questionnaire was used to record the incidence and severity of URI symptoms. There was no difference in the incidence of URIs between the vitamin D and placebo groups (48 URIs vs. 50 URIs, respectively, P=0.57). There was no difference in the duration or severity of URI symptoms between the vitamin D and placebo groups [5.4+/-4.8 days vs. 5.3+/-3.1 days, respectively, P=0.86 (95% CI for the difference in duration -1.8 to 2.1)]. The mean 25-hydroxyvitamin D level at baseline was similar in both groups (64.3+/ 25.4 nmol/l in the vitamin D group; 63.0+/-25.8 nmol/l in the placebo group; n.s.). After 12 weeks, 25-hydroxyvitamin D levels increased significantly to 88.5+/-23.2 nmol/l in the vitamin D group, whereas there was no change in vitamin D levels in the placebo group. There was no benefit of vitamin D3 supplementation in decreasing the incidence or severity of symptomatic URIs during winter. Further studies are needed to determine the role of vitamin D in infection. PMID- 19296871 TI - Investigation of an outbreak of Salmonella enterica serovar Newport infection. AB - A large outbreak of Salmonella enterica serotype Newport infection occurred in Northern Ireland during September and October 2004. Typing of isolates from patients confirmed that this strain was indistinguishable from that in concurrent outbreaks in regions of England, in Scotland and in the Isle of Man. A total of 130 cases were distributed unequally across local government district areas in Northern Ireland. The epidemic curve suggested a continued exposure over about 4 weeks. A matched case-control study of 23 cases and 39 controls found a statistically significant association with a history of having eaten lettuce in a meal outside the home and being a case (odds ratio 23.7, 95% confidence interval 1.4-404.3). This exposure was reported by 57% of cases. Although over 300 food samples were tested, none yielded any Salmonella spp. Complexity and limited traceability in salad vegetable distribution hindered further investigation of the ultimate source of the outbreak. PMID- 19296872 TI - The changing panorama of bacterial enteric infections. AB - We studied the age-specific population-based incidence of bacterial enteric infections caused by Shigella, Salmonella and Campylobacter, in Jerusalem. During 1990-2008, 32,408 cases were reported (incidence rate 232.1/100,000 per annum). The patterns of Shigella (47.4% of cases), Salmonella (34.4%) and Campylobacter (18.2%) infections evolved noticeably. Campylobacter rates increased from 15.0 to 110.8/100,000 per annum. Salmonella rates increased from 74.2 to 199.6/100,000 in 1995 then decreased to 39.4/100,000. Shigella showed an endemic/epidemic pattern ranging between 19.7 and 252.8/100,000. Most patients (75%) were aged <15 years; children aged <5 years comprised 56.4% of cases, despite accounting for only 12.9% of the population. Campylobacter was the predominant organism in infants aged <1 year and Shigella in the 1-4 years group. The hospitalization rates were: Shigella, 1.8%; Campylobacter, 2.3%; Salmonella, 6.9%. Infants were 2.2 times more likely to be hospitalized than children aged 1-14 years (P=0.001). Household transmission occurred in 21.2% of Shigella cases compared with 5% in the other bacteria. PMID- 19296873 TI - Climate variability and Ross River virus infections in Riverland, South Australia, 1992-2004. AB - Ross River virus (RRV) infection is the most common notifiable vector-borne disease in Australia, with around 6000 cases annually. This study aimed to examine the relationship between climate variability and notified RRV infections in the Riverland region of South Australia in order to set up an early warning system for the disease in temperate-climate regions. Notified data of RRV infections were collected by the South Australian Department of Health. Climatic variables and monthly river flow were provided by the Australian Bureau of Meteorology and South Australian Department of Water, Land and Biodiversity Conservation over the period 1992-2004. Spearman correlation and time-series adjusted Poisson regression analysis were performed. The results indicate that increases in monthly mean minimum and maximum temperatures, monthly total rainfall, monthly mean Southern Oscillation Index and monthly flow in the Murray River increase the likelihood, but an increase in monthly mean relative humidity decreases the likelihood, of disease transmission in the region, with different time-lag effects. This study demonstrates that a useful early warning system can be developed for local regions based on the statistical analysis of readily available climate data. These early warning systems can be utilized by local public health authorities to develop disease prevention and control activities. PMID- 19296874 TI - Nutritional modelling: distributions of salt intake from processed foods in New Zealand. AB - The salt content of processed foods is important because of the high intake of Na by most New Zealanders. A database of Na concentrations in fifty-eight processed foods was compiled from existing and new data and combined with 24 h diet recall data from two national nutrition surveys (5771 respondents) to derive salt intakes for seven population groups. Mean salt intakes from processed foods ranged from 6.9 g/d for young males aged 19-24 years to 3.5 g/d for children aged 5-6 years. A total of > or = 50 % of children aged 5-6 years, boys aged 11-14 years and young males aged 19-24 years had salt intakes that exceeded the upper limit for Na, calculated as salt (3.2-5.3 g/d), from processed foods only. Bread accounted for the greatest contribution to salt intake for each population group (35-43 % of total salt intake). Other foods that contributed 2 % or more and common across most age groups were sausage, meat pies, pizza, instant noodles and cheese. The Na concentrations of key foods have changed little over the 16-year period from 1987 to 2003 except for corned beef and whole milk that have decreased by 34 and 50 % respectively. Bread is an obvious target for salt reduction but the implication on iodine intake needs consideration as salt is used as a vehicle for iodine fortification of bread. PMID- 19296876 TI - Alveolar and cystic echinococcosis: towards novel chemotherapeutical treatment options. AB - Echinococcus granulosus and Echinococcus multilocularis are cestode parasites, of which the metacestode (larval) stages cause the neglected diseases cystic echinococcosis (CE) and alveolar echinococcosis (AE), respectively. The benzimidazoles albendazole and mebendazole are presently used for the chemotherapeutical treatment, alone or prior to and after surgery. However, in AE these benzimidazoles do not appear to be parasiticidal in vivo. In addition, failures in drug treatments as well as the occurrence of side-effects have been reported, leading to discontinuation of treatment or to progressive disease. Therefore, new drugs are needed to cure AE and CE. Strategies that are currently employed in order to identify novel chemotherapeutical treatment options include in vitro and in vivo testing of broad-spectrum anti-infective drugs or drugs that interfere with unlimited proliferation of cancer cells. The fact that the genome of E. multilocularis has recently been sequenced has opened other avenues, such as the selection of novel drugs that interfere with the parasite signalling machinery, and the application of in silico approaches by employing the Echinococcus genome information to search for suitable targets for compounds of known mode of action. PMID- 19296875 TI - Independent and interactive effects of plant sterols and fish oil n-3 long-chain polyunsaturated fatty acids on the plasma lipid profile of mildly hyperlipidaemic Indian adults. AB - The present study was designed to evaluate the independent and interactive effects of a once-a-day yoghurt drink providing 2 g plant sterols/d and capsules providing 2 g fish oil n-3 long-chain (LC) PUFA/d on plasma lipids, apolipoproteins and LDL particle size. Following a 2-week run-in period, 200 mildly hypercholesterolaemic Indian adults aged 35-55 years were randomised into one of four groups of a 2 x 2 factorial, double-blind controlled trial. The 4 week treatments consisted of (1) control yoghurt drink and control capsules, (2) control yoghurt drink and fish oil capsules, (3) plant sterol-enriched yoghurt drink and control capsules, or (4) plant sterol-enriched yoghurt drink and fish oil capsules. Blood was drawn before and after the 4-week intervention. Changes in health status, lifestyle and dietary habits, and daily compliance were recorded. The main effects of plant sterols were a 4.5 % reduction in LDL cholesterol and a 15 % reduction in TAG without a significant change in HDL cholesterol. Overall, fish oil n-3 LC-PUFA did not significantly affect cholesterol concentrations but reduced TAG by 15 % and increased HDL-cholesterol by 5.4 %. The combination significantly lowered TAG by 15 % v. control. No significant interaction between plant sterols and n-3 LC-PUFA was observed on plasma cholesterol concentrations. In conclusion, once-a-day intake of 2 g plant sterols/d in a yoghurt drink, 2 g fish oil n-3 LC-PUFA/d in capsules, and their combination had beneficial effects on the lipid profile of mildly hypercholesterolaemic Indian adults. The potent hypotriacylglycerolaemic effect of plant sterols observed in the present study and this population warrants additional investigation. PMID- 19296877 TI - Bird schistosome diversity in Iceland: molecular evidence. AB - During the years 2002-2007, Icelandic freshwater snails and birds from different orders were examined for bird schistosomes. Only the snail Radix peregra and anatid birds proved to be infected. In total, 32 samples of bird schistosome cercariae from seven localities and four samples of adults of Anas platyrhynchos and Mergus serrator from two localities were used for sequencing of the internal transcibed spacer (ITS) region of rDNA. Based on the sequence and position in the phylogenetic tree, five species/genotypes of bird schistosomes were identified. Three of them correspond to the sequences in GenBank for Trichobilharzia franki, Trichobilharzia sp. JR-2007 from Anas penelope (isolates Pl7 and Pl10) and schistosomatid sp. JR-2004 from Physa fontinalis. The other two species/genotypes belong to the genus Trichobilharzia, but their further determination was not possible due to lack of data. All of these species/genotypes develop in R. peregra, probably the only suitable intermediate host species in Iceland. As T. franki and schistosomatid sp. JR-2004 were previously reported from other snail species, the findings from Iceland proved that, under some circumstances, bird schistosomes are able to adapt to a different snail species; some of them can even adapt to a snail belonging to a different gastropod family. High diversity of bird schistosomes in Iceland is probably linked with migratory flyways of birds, as several bird species from Iceland overwinter in western Europe as well as in North America. PMID- 19296878 TI - Why are you smiling at me? Social functions of enjoyment and non-enjoyment smiles. AB - In three experiments, we investigated the spontaneous attention of perceivers to the nature of targets' facial expressions, specifically whether they were displaying an enjoyment or a non-enjoyment smile. Further, we investigated the social functions of sensitivity to smile type and the consequences of such sensitivity for subsequent interactions. Results demonstrated that perceivers did indeed spontaneously attend to smile type, especially in situations where issues of trust or cooperation were made salient. Further, this sensitivity had an impact both on the evaluations of the target individuals and the cooperative behaviour of individuals towards those displaying enjoyment and non-enjoyment smiles. Participants evaluated individuals displaying enjoyment smiles more positively than those displaying non-enjoyment smiles and had higher rates of cooperation with those displaying enjoyment smiles. These results are discussed in terms of the social functions of facial expressions. PMID- 19296879 TI - A sugar transporter regulates serum urate levels: implications for prevention and management of hyperuricemia in gout. PMID- 19296880 TI - Genetics of systemic sclerosis-associated pulmonary arterial hypertension: recent progress and current concepts. AB - Pulmonary arterial hypertension (PAH) is a devastating complication of systemic sclerosis (SSc). Although there are screening tests for PAH, such as carbon monoxide diffusing capacity and Doppler echocardiography, these tests are far from perfect, patients are still identified late in the course of disease, and the underlying mechanisms of SSc-associated PAH are poorly understood. Understanding the genetic differences between those patients with SSc who do and do not develop PAH may improve our ability to identify and treat patients earlier in the course of disease. To date, only a few candidate gene association studies and one gene expression analysis have investigated the genetics of PAH in SSc. These studies highlight the important opportunities and challenges for exploration of genetics of complex traits. By appreciating the optimal conduct of genetic studies, along with the role of bioinformatics resources such as the International HapMap database, investigators and clinicians will be better equipped to understand the genetics of SSc-associated PAH. PMID- 19296881 TI - Current developments in pediatric systemic sclerosis. AB - Juvenile systemic sclerosis is an orphan disease with an incidence of around 0.05 per 100,000 children. The mean age of onset is approximately 8 years, and 90% of pediatric patients have a diffuse subset. The organ involvement of pediatric patients differs from adult patients. Survival rates are significantly better in pediatric patients than in adult patients. Most patients who die in the first 5 years of the disease have a diffuse subset. Interestingly, the disease subset shifts when pediatric patients are reviewed in adult cohorts of systemic sclerosis patients, in which only around 40% of patients have diffuse subset. The pediatric-onset patients still have a low prevalence of anticentromere antibodies. There is a larger group of patients with overlap features in juvenile systemic sclerosis than in adult cohorts. There are also noticeable differences in organ involvement. PMID- 19296882 TI - Is scleroderma a vasculopathy? AB - Described as an autoimmune collagen vascular disease, the most striking feature of scleroderma may be a systemic vasculopathy. This vasculopathy includes characteristic noninflammatory macrovascular and microvascular changes with dramatic and possibly occlusive formation of a thickened neointima. Scleroderma vessels also have an unusual endothelial phenotype, with loss of normal markers including vascular endothelial (VE)-cadherin. These endothelial cells express type 1 interferon and regulator of G protein signaling 5 (RGS5), two molecules associated with vascular rarefaction. These genes may be important because tissue is hypoxic with high levels of vascular endothelial growth factor (VEGF), especially early in the disease. The combination of VEGF and rarefaction is not necessarily paradoxical. VEGF-mediated angiogenesis creates labile vessels that may not survive unless the vessel acquires a smooth muscle coat. The combination of interferon and RGS5 is consistent with an antiangiogenic phenotype. We offer a hypothesis that places vascular injury at the center of this disease and also suggest possible clinical approaches for arresting and/or reversing the disease. PMID- 19296883 TI - Current concepts in disease-modifying therapy for systemic sclerosis-associated interstitial lung disease: lessons from clinical trials. AB - Interstitial lung disease (ILD) is the leading cause of mortality in patients with systemic sclerosis (SSc), which is also known as scleroderma. Two randomized clinical trials in patients with SSc-related ILD have shown that oral or intravenous cyclophosphamide is associated with modest but significant or near significant improvements in lung function, dyspnea, and physical function. In addition, the Scleroderma Lung Study and an observational study showed that baseline forced vital capacity less than 70% and moderate fibrosis on thoracic high-resolution CT are predictors of response to cyclophosphamide therapy and/or survival, whereas active alveolitis on bronchoalveolar lavage is not. Newer therapies for SSc patients with ILD include mycophenolate mofetil, tyrosine kinase inhibitors (imatinib, dasatinib), and anti-interleukin-13 monoclonal antibody. Several uncontrolled trials have reported favorable results of mycophenolate mofetil in SSc-related ILD. A randomized double-blind controlled trial by the Scleroderma Lung Study Research Group is currently comparing the efficacy and safety of mycophenolate mofetil versus oral cyclophosphamide. PMID- 19296884 TI - Tissue stiffness, latent TGF-beta1 activation, and mechanical signal transduction: implications for the pathogenesis and treatment of fibrosis. AB - Tissue stiffening is a predominant feature of fibrosis and it obstructs organs whose mechanical properties are important for their function, such as the heart, lung, skin, and vessels. Stiff scar tissue further modulates the character of the healthy residing cells by driving the differentiation of a variety of precursor cells into fibrogenic myofibroblasts. This mechanical cue for myofibroblast differentiation establishes a vicious cycle because the excessive extracellular matrix-secreting and remodeling activities of myofibroblasts are cause and effect of further connective tissue contracture and stiffening. The second pivotal factor inducing myofibroblast development is transforming growth factor-beta1. Recent findings suggest that transforming growth factor-beta1 activity is partly controlled by myofibroblast contractile forces and tissue stiffness. This discovery opens new paths to prevent progression of fibrosis by specifically interfering with the stress perception and transmission mechanisms of the myofibroblast. PMID- 19296885 TI - Calcifying tendinitis of the shoulder: advances in imaging and management. AB - Calcifying tendonitis of the shoulder is a common, acute or chronic, painful disorder characterized by calcifications in the rotator cuff tendons. A natural cycle exists during which the tendon repairs itself. In chronic calcific tendonitis, however, this cycle is blocked at one of the healing stages. Because chronic presentation with exacerbations is usual, initial treatment should be conservative, including rest, physical therapy, nonsteroidal anti-inflammatory drugs, and, in later stages, subacromial infiltration with corticosteroids. Surgery is recommended when conservative treatment fails. This article discusses advances in imaging and medical, physical, and surgical management, as well as current evidence for the treatment of calcifying tendonitis of the shoulder. PMID- 19296886 TI - A critical reappraisal of allopurinol dosing, safety, and efficacy for hyperuricemia in gout. AB - Allopurinol, the first-line drug for serum urate-lowering therapy in gout, is approved by the US Food and Drug Administration for a dose up to 800 mg/d and is available as a low-cost generic drug. However, the vast majority of allopurinol prescriptions are for doses < or = 300 mg/d, which often fails to adequately treat hyperuricemia in gout. This situation has been promoted by longstanding, non-evidence-based guidelines for allopurinol use calibrated to renal function (and oxypurinol levels) and designed, without proof of efficacy, to avoid allopurinol hypersensitivity syndrome. Severe allopurinol hypersensitivity reactions are not necessarily dose-dependent and do not always correlate with serum oxypurinol levels. Limiting allopurinol dosing to < or = 300 mg/d suboptimally controls hyperuricemia and fails to adequately prevent hypersensitivity reactions. However, the long-term safety of elevating allopurinol dosages in chronic kidney disease requires further study. The emergence of novel urate-lowering therapeutic options, such as febuxostat and uricase, makes timely this review of current allopurinol dosing guidelines, safety, and efficacy in gout hyperuricemia therapy, including patients with chronic kidney disease. PMID- 19296887 TI - Point: Hydroxyapatite crystal deposition is intimately involved in the pathogenesis and progression of human osteoarthritis. AB - The cause of osteoarthritis (OA), the most common form of arthritis, is most likely multifactorial. No drug exists to slow the progression or reverse OA disease progression. Ample data support a key role of calcium-containing crystals, such as hydroxyapatite, in OA pathogenesis. The presence of these crystals, far higher in OA than in any other form of arthritis, correlates with the degree of radiographic degeneration. Calcium-containing crystals have potent biologic effects in vitro that emphasize their pathogenic potential. OA associated matrix and chondrocyte alterations play an intimate role in the crystal deposition process. A major difficulty has been the lack of a simple technique for crystal identification in affected joints. Enhanced effort is needed to establish calcium-containing crystals as a therapeutic target in OA, as current data suggest an intimate association in its pathogenesis and progression. PMID- 19296888 TI - Counterpoint: Hydroxyapatite crystal deposition is not intimately involved in the pathogenesis and progression of human osteoarthritis. AB - The association of hydroxyapatite deposition with osteoarthritis pathogenesis and progression remains controversial, even after decades of study. Hydroxyapatite crystals are found in osteoarthritis in advanced disease only. Even then, hydroxyapatite crystals are found in such small amounts that special analytical techniques are required to detect the crystals. Further, the osteoarthritic joint fluid appears noninflammatory, suggesting that such hydroxyapatite crystals have very small effect on the pathologic process. Formal histopathologic evidence is scant, but hydroxyapatite crystal deposition within osteoarthritic synovium or cartilage is a rare event. Hydroxyapatite crystals may be present at end-stage osteoarthritis, but in insufficient amounts to contribute significantly to osteoarthritis pathogenesis or progression. This review critically examines the evidence from osteoarthritic synovial fluids, imaging, and histopathology to determine whether the well-characterized in vitro cellular reactions to hydroxyapatite apply to the pathogenesis of human osteoarthritis. PMID- 19296889 TI - Quality of life and quality of care for patients with gout. AB - Significant pain, activity limitation, and disability in patients with acute and chronic gouty arthritis lower health-related quality of life. Although many effective therapies are available for gouty arthritis, medication errors are common. One goal of therapy is to reduce the frequency of gout flares by lowering serum uric acid. Further, evidence suggests that the quality of care provided to patients with gout may also impact health-related quality of life. This article reviews evidence concerning quality of care and quality of life for patients with gout. PMID- 19296890 TI - A comparison of females and males for antiperspirant efficacy and sweat output. PMID- 19296891 TI - Moisturizing and anti-sebum secretion effects of cosmetic application on human facial skin. AB - For human skin, high water content and low sebum secretion are considered to be main features of fair skin. To explore the proper personal care regimen for facial skin, we investigated the change of skin physiologic parameters after cosmetic application by measuring the skin water content, transepidermal water loss, and skin sebum secretion on facial skin before and after the cosmetic application using the Corneometer, Tewameter, and Sebumeter, respectively. The results indicated that the cosmetics application kept a higher water content and a lower transepidermal water loss, and at the same time, a lower sebum secretion 4 h and 8 h after the cosmetic application, compared with those before it. The situation was maintained in the succeeding three-week continuous use of the cosmetics. It could be concluded that the cosmetic application on human facial skin might provide some moisturizing effect and at the same time an anti-sebum effect, which favors the maintenance of good skin physiological function after applying skin care products. Our results might provide a scientific personal care regimen for human facial skin to prompt the balance for the hydrolipid film on skin. PMID- 19296892 TI - Skin moisturization by hydrogenated polyisobutene--quantitative and visual evaluation. AB - Hydrogenated polyisobutene (HP) is used in topically applied cosmetic/personal care formulations as an emollient that leaves a pleasing skin feel when applied, and rubbed in after application. This effect, although distinguishable to the user, is difficult to define and quantify. Recognizing that some of the physical properties of HP such as film formation and wear resistance may contribute, in certain mechanisms, to skin moisturization, we designed a short-term pilot study to follow changes in skin moisturization. HP's incorporation into an o/w emulsion at 8% yielded increased viscosity and reduced emulsion droplet size as compared to the emollient ester CCT (capric/caprylic triglyceride) or a control formulation. Quantitative data indicate that application of the o/w emulsion formulation containing either HP or CCT significantly elevated skin moisture content and thus reduced transepidermal water loss (TEWL) by a maximal approximately 33% against the control formulation within 3 h and maintained this up to 6 h. Visual observation of skin treated with the HP-containing formulation showed fine texture and clear contrast as compared to the control or the CCT formulation, confirming this effect. As a result of increased hydration, skin conductivity, as measured in terms of corneometer values, was also elevated significantly by about tenfold as early as 20 min after HP or CCT application and was maintained throughout the test period. Throughout the test period the HP formulation was 5-10% more effective than the CCT formulation both in reduction of TEWL as well as in increased skin conductivity. Thus, compared to the emollient ester (CCT), HP showed a unique capability for long-lasting effect in retaining moisture and improving skin texture. PMID- 19296893 TI - A rapid method to clinically assess the effect of an anti-acne formulation. AB - Historically, clinical evaluation of acne treatment has been based on direct visual assessment and the counting of lesions over a period of several weeks of treatment. However, with advancing technology there has been ever-increasing speed in the effectiveness of these treatments. To successfully assess these faster treatments, acne pathology needs to be evaluated in a shorter time frame. The object of these studies was to develop techniques to evaluate individual acne lesions in a shorter time frame and to assess speedier treatment technologies. Ten healthy volunteers with acne lesions on their upper backs were recruited for the study. Two inflamed acne lesions were selected for each treatment, along with lesions to be left untreated, on each volunteer. Each lesion was marked, photographed, and visually graded. A skin surface microscope (Scopeman) was used to visualize size and to grade the lesions by two experts every day for five days. The sites were treated once a day for the course of the study. There was a remarkable reduction in the size and erythema of acne lesions after treatment with the acne formulation as compared to the untreated and vehicle-treated lesions. Individual lesions, both treated and untreated, appeared resolved in 14 days. This resolution can be noticeably accelerated by topical treatments. We have developed a simple and faster clinical method to evaluate the effects of topical anti-acne technology. PMID- 19296894 TI - Deposition of 18-MEA onto alkaline-color-treated weathered hair to form a persistent hydrophobicity. AB - A technology for the deposition of a persistent hydrophobicity to alkaline-color treated weathered hair surfaces using 18-MEA (18-methyleicosanoic acid) is presented. Two approaches were examined in order to make 18-MEA bind tightly to the alkaline-color-treated weathered hair surface. One was to apply 18-MEA as an acid form and the other was to apply 18-MEA as a salt or complex. It was found that the combination of 18-MEA with specific cationic surfactants [stearoxypropyldimethylamine (SPDA) and docosyldimethylamine (DSDA)] makes the alkaline-color-treated weathered hair surface hydrophobic and that its hydrophobicity is maintained even after shampooing. Characterization of adsorbed layers of 18-MEA/SPDA on a mica surface, as a possible hydrophilic surface model, was performed using atomic force microscopy (AFM) and angle-resolved X-ray photoelectron spectroscopy (AR-XPS). The results revealed that 18-MEA/SPDA formed a layer with high wear resistance, with an alkyl chain, the hydrophobic moiety, oriented at an angle of around 25 degrees to the air interface. PMID- 19296901 TI - TaqMan real-time PCR quantification strategy of CYP2D6 gene copy number for the LightCycler 2.0. AB - BACKGROUND: The metabolism of many therapeutic drugs depends on the presence and activity of CYP2D6 enzymes. Poor or ultrarapid metabolism may lead to adverse drug effects and lack of therapeutic efficacy. Determining the CYP2D6 gene copy number (GCN) together with SNP genotyping allows predicting the CYP2D6 phenotype and may be beneficial for patients. Efficient TaqMan real-time PCR assays have been developed for this specification but are limited to the Abi Prism system and lack extensive data to demonstrate reliable application for routine purposes. MATERIALS AND METHODS: We established two TaqMan real-time PCR assays to quantify CYP2D6 GCN on the LightCycler 2.0 platform. With albumin as internal control, one assay targets the exon 9 region of the CYP2D6; the other the intron 6. RESULTS: In 617 samples there is a 99.4% (exon 9 method) and 95.6% (intron 6 method) correlation compared to standard methods. Analyzing deviant results offer indications for polymorphisms such as CYP2D616 and exon 9 gene conversions. CONCLUSION: Established TaqMan real-time PCR assays to determine CYP2D6 GCN on the LightCycler 2.0 are reliable and may be used in the routine. Comparing deviant results, these assays may even allow the screening for rare polymorphism. PMID- 19296902 TI - De novo deletion at D13S317 locus: a case of paternal-child allele mismatch identified by microsatellite typing. PMID- 19296903 TI - Haptoglobin phenotypes in cervical cancer: decreased risk for Hp2-2 individuals. PMID- 19296904 TI - Preference for dietary fat induced by release of beta-endorphin in rats. AB - AIMS: To determine whether beta-endorphin contributes to the ingestion of and preference for dietary oil, we examined the relationship between the dynamics of beta-endorphin, before and after the ingestion of corn oil, and the intake volume of corn oil. MAIN METHODS: Rats were offered 5% corn oil for 20 min for 5 consecutive days so they could acquire a preference for corn oil. On day 6, seven groups of rats were presented with the oil for defined time periods, and we measured the beta-endorphin levels in the serum and cerebrospinal fluid (CSF) before and after the presentation of corn oil as well as the consumed volume of corn oil at defined time points. KEY FINDINGS: Beta-endorphin levels in serum and CSF were significantly increased 15 min after the ingestion of corn oil, followed by a rapid decrease and maintenance at the basal level throughout the rest of the experimental period. The intake of corn oil was the lowest in the time period of 15-30 min, when the beta-endorphin level reached a peak value. The intake volume of corn oil might be inversely correlated with beta-endorphin levels in serum and CSF. The pretreatment of naloxone, an antagonist of the opioid receptor, decreased the initial licking rate for corn oil and increased the latency for corn oil in the licking test. SIGNIFICANCE: The beta-endorphin was rapidly released after oil ingestion, which contributed to the hedonic preference and ingestive behavior for fat. PMID- 19296905 TI - Consummatory successive negative and anticipatory contrast effects in inbred Roman rats. AB - Three experiments were designed to study the performance of female inbred Roman High-(RHA-I) and Low-(RLA-I) Avoidance rats in a consummatory task in which successive negative (cSNC) and anticipatory contrast (cANC) effects were induced by changing the concentration of the sucrose solution used as reward. Both RHA-I and RLA-I rats showed a significant suppression of drinking (cSNC) when they were exposed to 32% sucrose in preshift phase and 4% in postshift phase, in comparison to RHA-I and RLA-I control groups always exposed to 4% sucrose (Experiment 1). By contrast, when the preshift-postshift reward discrepancy was reduced from 32-4 to 22-4 in Experiment 2, both strains showed a suppression of fluid intake on the first postshift trial, whereas only the more emotional RLA-I strain maintained this suppression on subsequent days. Finally, no consumption differences were observed between RHA-I and RLA-I rats exposed to brief daily access to 4% sucrose followed by access to 22% sucrose (cANC, Experiment 3). These results suggest that differences between the Roman rat strains in consummatory contrast paradigms can only be observed when the sucrose solution manipulation implies an incentive loss-attenuated emotional situation, but not when it occurs in an anticipatory manner. PMID- 19296906 TI - The passive coping Roman Low Avoidance rat, a non-obese rat model for insulin resistance. AB - The aim of the study was develop to an animal model that links coping style to insulin resistance. We hypothesized that the psychogenetically selected Roman Low Avoidance (RLA) rats may serve as such a model. To test this hypothesis, we submitted both RLA and Roman High avoidance (RHA) rats to a series of intravenous glucose tolerance tests (IVGTT). These IVGTT were followed by post mortem metabolic characterization of the selection lines. It was found that plasma insulin levels are markedly elevated in the passively coping RLA rat, both in baseline conditions and during the intravenous glucose tolerance tests. The elevation in plasma insulin was accompanied with increased levels of plasma corticosterone, FFA, leptin and triglycerides but not by changes in body weight. We conclude that the passive, highly emotional RLA rat is metabolically different from both the RHA rat and the standard control Wistar rat and may serve as a non obese animal model for insulin resistance. PMID- 19296907 TI - Mammalian target of rapamycin complex 1 (mTORC1) signaling in energy balance and obesity. AB - Energy balance is guaranteed by a complex circuitry that in the brain, and in the hypothalamus in particular, integrates and coordinates several types of signals, including hormones and nutrients, so to match energy expenditure to energy needs. Similar to individual cells, the hypothalamus also profits from intracellular pathways known to work as fuel sensors to maintain energy balance. The mammalian target of rapamycin complex 1 (mTORC1) pathway has been recently implicated in such function, due to its ability to integrate nutrient and hormonal signals to control food intake and body weight. This review therefore describes recent advances made in understanding the role of the hypothalamic mTORC1 pathway in energy balance regulation and its possible contribution to the metabolic dysregulation associated with diet-induced obesity. PMID- 19296908 TI - Fecal hormone variation during prolonged social interaction in male Tscheskia triton. AB - Agonistic behavior is crucial for conspecific members to maintain a social hierarchy, optimum population density, and high fitness. It is known that agonistic behavior and social ranking often interact with hormones such as testosterone (T) and glucocorticoids (GCs). The challenge hypothesis states that T levels in males are promoted by the agonistic behaviors of other males and has been widely testified in many taxa of vertebrates, even in humans, but seldom attempted in rodents. Here, we examined how fecal T and corticosterone (CORT) concentrations changed during prolonged social conflict in male greater long tailed hamsters (Tscheskia triton). Dyads were subjected to 5 min staged encounters daily for 15 days during which agonistic and social behaviors were recorded and fecal hormone concentrations were determined by radioimmunoassay. Our results showed that pairwise male hamsters developed overt and stable dominant-subordinate relationships rapidly and that the agonistic behavior decreased over the course of the experiment. Dominant males exhibited more frequent flank marking and locomotion and shorter latency to initial attack than their subordinate counterparts. Testosterone levels were significantly increased in both dominant and subordinate males during early encounters, but T and CORT levels were higher in subordinate males. After five encounters, we found no difference between hormone levels and behavior for all males, implying some kind of behavioral and physiological habituation. This complex pattern of hormonal change during social conflict is discussed and correlations between behavioral and physiological habituation are hypothesized. PMID- 19296909 TI - The effect of postnatal exposure to noise on sound level processing by auditory cortex neurons of rats in adulthood. AB - Most people are exposed daily to some level and duration of environmental noise. The aim of the present study was to determine the effect of postnatal exposure to a moderate level of noise on sound level processing by neurons in the primary auditory cortex of rats in adulthood. The cortical neuron response to sound stimuli was investigated in three groups of rats. Two groups, either in the critical period of postnatal hearing development or in adulthood, were exposed to 80 dB SPL interrupted white noise for 8 h/day for 2 weeks. The control group consisted of adult rats that were not exposed to the white noise. Seven weeks later, the minimum threshold, the first spike latency, the dynamic range and the slope of the rate-level functions of cortical neuron response to a sound stimulus were determined. The cortical neurons in young rats exposed to the noise had a significantly higher minimum threshold, a longer first spike latency, a shorter dynamic range and a bigger slope in rate-level functions compared with the control group. The group in which adult rats were exposed to the white noise, however, did not have a significant change of sound level processing by the auditory cortical neurons. These results demonstrated that young rats were more susceptible to noise exposure affecting the cortical neuron processing of sound levels. PMID- 19296910 TI - The effects of long-term honey, sucrose or sugar-free diets on memory and anxiety in rats. AB - Sucrose is considered by many to be detrimental to health, giving rise to deterioration of the body associated with ageing. This study was undertaken to determine whether replacing sucrose in the diet long-term with honey that has a high antioxidant content could decrease deterioration in brain function during ageing. Forty-five 2-month old Sprague Dawley rats were fed ad libitum for 52 weeks on a powdered diet that was either sugar-free or contained 7.9% sucrose or 10% honey (which is the equivalent amount of sugar). Anxiety levels were assessed using an Elevated Plus Maze, whilst a Y maze and an Object Recognition task were used to assess memory. Locomotor activity was also measured using an Open Field task to ensure that differences in activity levels did not bias results in the other tasks. Anxiety generally decreased overall from 3 to 12 months, but the honey-fed rats showed significantly less anxiety at all stages of ageing compared with those fed sucrose. Honey-fed animals also displayed better spatial memory throughout the 12-month period: at 9 and 12 months a significantly greater proportion of honey-fed rats recognised the novel arm as the unvisited arm of the maze compared to rats on a sugar-free or sucrose-based diet. No significant differences among groups were observed in the Object Recognition task, and there appeared to be no differences in locomotor activity among groups at either 6 or 12 months. In conclusion, it appears that consumption of honey may reduce anxiety and improve spatial memory in middle age. PMID- 19296911 TI - Mimicking catatonic symptoms: a head injury induced acute hyponatremia complicated by rhabdomyolysis in a patient with schizophrenia. PMID- 19296912 TI - The role of leptin and cortisol in hyperactivity in patients with acute and weight-recovered anorexia nervosa. AB - INTRODUCTION: In food-restricted rats, leptin as well as corticotropin releasing factor attenuate semistarvation-induced hyperactivity (SIH). Results from studies in patients with anorexia nervosa (AN) showed an association between excessive physical activity (PA) and leptin. One recent report suggests a role for cortisol in PA. In this study, we assessed the relationships between PA and both, cortisol and leptin levels at the same time in patients with acute anorexia nervosa (acAN) in comparison to recovered patients (recAN). METHODS: Plasma leptin, plasma cortisol, body mass index (BMI), and expert-ratings of qualities of PA were assessed in 36 acAN patients, 27 recAN patients and 44 healthy control woman (HCW). Regression analyses were used to predict PA using BMI, leptin and cortisol levels as predictor variables. RESULTS: Leptin levels but not cortisol significantly contributed to the prediction of PA in acAN. In recAN PA was not elevated and not related to endocrine parameters but correlated positively with core eating disorder symptoms. CONCLUSIONS: Our work lends support to the proposed inverse association between peripheral leptin levels and excessive physical activity in AN. This relationship is specific to the state of semistarvation. The role of additional mediators remains to be clarified. PMID- 19296913 TI - Phosphorylation of the activation loop tyrosines is required for sustained Syk signaling and growth factor-independent B-cell proliferation. AB - The Syk kinase is regarded as a promising target for the treatment of antigen driven B-cell malignancies, considering its essential role in propagating antigenic stimuli through the B-cell receptor (BCR). In certain common B-cell malignancies Syk is activated even in the absence of BCR engagement, suggesting a wider role for this kinase in lymphomagenesis. In this paper, we have profiled molecular differences between BCR-induced and constitutive Syk activation in terms of phosphorylation of regulatory tyrosine residues, downstream signaling properties and capacity to sustain B-cell proliferation. Analysis of primary chronic lymphocytic leukemia B-cells and diffuse large B-cell lymphoma cell lines revealed that constitutive and BCR-induced Syk activation differ with respect to the phosphorylation status of the regulatory tyrosines at positions 352 and 525/526, with only the first site being phosphorylated in the case of constitutive and both sites in the case of BCR-induced Syk activation. Syk phosphorylated only on Y352 is capable of downstream signaling, as evidenced by experiments with a phosphomimetic mutant in which the activation loop tyrosines (YY525/526) were replaced with phenylalanines. However, phosphorylation at YY525/526 was shown to significantly increase the enzymatic activity of Syk and to be required for sustained PLCgamma2, Akt and ERK signaling as well as B-cell transformation. These data demonstrate that constitutively active Syk and Syk activated by BCR crosslinking represent separate stages of Syk activation with distinct signaling properties and transforming capacities. PMID- 19296914 TI - Arf GAP2 is positively regulated by coatomer and cargo. AB - Arf GAP2 is one of four Arf GAPs that function in the Golgi apparatus. We characterized the kinetics of Arf GAP2 and its regulation. Purified Arf GAP2 had little activity compared to purified Arf GAP1. Of the potential regulators we examined, coatomer had the greatest effect, stimulating activity one to two orders of magnitude. The effect was biphasic, with half-maximal activation observed at 50 nM coatomer and activation peaking at approximately 150 nM coatomer. Activation by coatomer was greater for Arf GAP2 than has been reported for Arf GAP1. The effects of phosphoinositides and changes in vesicle curvature on GAP activity were small compared to coatomer; however, both increased coatomer dependent activity. Peptides from p24 cargo proteins increased Arf GAP2 activity by an additional 2- to 4-fold. The effect of cargo peptide was dependent on coatomer. Overexpressing the cargo protein p25 decreased cellular Arf1*GTP levels. The differential sensitivity of Arf GAP1 and Arf GAP2 to coatomer could coordinate their activities. Based on the common regulatory features of Arf GAP1 and 2, we propose a mechanism for cargo selection in which GTP hydrolysis triggered by cargo binding to the coat protein is coupled to coat polymerization. PMID- 19296915 TI - VGD and MLD-motifs containing heterodimeric disintegrin viplebedin-2 from Vipera lebetina snake venom. Purification and cDNA cloning. AB - We have previously demonstrated that the fibrinolytic enzyme lebetase is synthesized with disintegrin-like domain that is cleaved posttranslationally (Siigur et al., 1996). Now we isolated a heterodimeric disintegrin viplebedin-2 containing this disintegrin-like part from Vipera lebetina venom using size exclusion chromatography on Sephadex G-100 sf and HPLC on C18 column. The molecular masses of viplebedin-2 and tryptic peptides from both chains of viplebedin-2 were determined by MALDI-TOF mass spectrometry. Using cDNA library of the venom gland of a single V. lebetina turanica snake the viplebedin-2 coding cDNAs were cloned and sequenced. Viplebedin-2 chains are synthesized from two different genes. One chain, containing VGD sequence in disintegrin loop, is synthesized as a disintegrin-like part of the PII-type metalloprotease, lebetase. The other chain, containing MLD sequence in disintegrin loop, is synthesized from the gene without metalloproteinase domain. Two polyadenylation signal sequences have been found in MLD sequence coding chain precursor cDNAs. Viplebedin-2 dose dependently inhibited adhesion of platelets to immobilized collagen and inhibited collagen-induced platelet aggregation. PMID- 19296916 TI - US stimulus bill provides $10 billion boost to NIH. PMID- 19296917 TI - Evidence-based practice for stroke. PMID- 19296918 TI - Assisted suicide: a matter of life and death. PMID- 19296919 TI - Ruth Bonita: passionate about prevention. PMID- 19296920 TI - The borderland of epilepsy: clinical and molecular features of phenomena that mimic epileptic seizures. AB - Paroxysmal losses of consciousness and other episodic neurological symptoms have many causes. Distinguishing epileptic from non-epileptic disorders is fundamental to diagnosis, but even this basic dichotomy is often challenging and is certainly not new. In 1907, the British neurologist William Richard Gowers published his book The Border-land of Epilepsy in which he discussed paroxysmal conditions "in the border-land of epilepsy-near it, but not of it" and their clinical differentiation from epilepsy itself. Now, a century later, we revisit the epilepsy borderland, focusing on syncope, migraine, vertigo, parasomnias, and some rarer paroxysmal disorders. For each condition, we review the clinical distinction from epileptic seizures. We then integrate current understanding of the molecular pathophysiology of these disorders into this clinical framework. This analysis shows that, although the clinical manifestations of paroxysmal disorders are highly heterogeneous, striking similarities in molecular pathophysiology are seen among many epileptic and non-epileptic paroxysmal phenomena. PMID- 19296921 TI - Neuroinflammation in Parkinson's disease: a target for neuroprotection? AB - Parkinson's disease is characterised by a slow and progressive degeneration of dopaminergic neurons in the substantia nigra. Despite intensive research, the cause of the neuronal loss in Parkinson's disease is poorly understood. Neuroinflammatory mechanisms might contribute to the cascade of events leading to neuronal degeneration. In this Review, we describe the evidence for neuroinflammatory processes from post-mortem and in vivo studies in Parkinson's disease. We further identify the cellular and molecular events associated with neuroinflammation that are involved in the degeneration of dopaminergic neurons in animal models of the disease. Overall, available data support the importance of non-cell-autonomous pathological mechanisms in Parkinson's disease, which are mostly mediated by activated glial and peripheral immune cells. This cellular response to neurodegeneration triggers deleterious events (eg, oxidative stress and cytokine-receptor-mediated apoptosis), which might eventually lead to dopaminergic cell death and hence disease progression. Finally, we highlight possible therapeutic strategies (including immunomodulatory drugs and therapeutic immunisation) aimed at downregulating these inflammatory processes that might be important to slow the progression of Parkinson's disease. PMID- 19296923 TI - [Gene profiling and classification of adrenocortical tumors]. PMID- 19296924 TI - A matter of dosage: SF-1 in adrenocortical development and cancer. PMID- 19296922 TI - Preconditioning and tolerance against cerebral ischaemia: from experimental strategies to clinical use. AB - Neuroprotection and brain repair in patients after acute brain damage are still major unfulfilled medical needs. Pharmacological treatments are either ineffective or confounded by adverse effects. Consequently, endogenous mechanisms by which the brain protects itself against noxious stimuli and recovers from damage are being studied. Research on preconditioning, also known as induced tolerance, over the past decade has resulted in various promising strategies for the treatment of patients with acute brain injury. Several of these strategies are being tested in randomised clinical trials. Additionally, research into preconditioning has led to the idea of prophylactically inducing protection in patients such as those undergoing brain surgery and those with transient ischaemic attack or subarachnoid haemorrhage who are at high risk of brain injury in the near future. In this Review, we focus on the clinical issues relating to preconditioning and tolerance in the brain; specifically, we discuss the clinical situations that might benefit from such procedures. We also discuss whether preconditioning and tolerance occur naturally in the brain and assess the most promising candidate strategies that are being investigated. PMID- 19296925 TI - Animal models of primary aldosteronism. AB - With an average prevalence of 25% hypertension is one of the leading chronic diseases in westernized countries. Recent epidemiological data indicate a high proportion of patients with secondary hypertension due to primary aldosteronism that accounts for up to 8-12% of cases. Primary aldosteronism is caused by autonomous secretion of aldosterone by the adrenal cortex which results in hypertension with clinically, biochemically and therapeutically distinct features. With the exception of the small proportion of patients with familial hyperaldosteronism type I, the underlying genetic and molecular basis of this common disease is largely unknown. In this situation mouse models with targeted genetic modification can be utilized to define functional relevance of predefined candidate genes that are known or suspected to be involved in the regulation of aldosterone secretion. Moreover, animal models can aid in the identification of novel gene products that have not yet been identified to play a role in primary aldosteronism. This review will provide a brief overview on the animal models currently available for primary aldosteronism and describe in vivo screening strategies that are likely to provide insight in molecular and genetic mechanisms involved in autonomous aldosterone secretion. PMID- 19296926 TI - Biochemical diagnosis of pheochromocytoma and paraganglioma. PMID- 19296927 TI - [Papillary adenocarcinoma on ectopic thyroid tissue]. AB - OBJECTIVES: To report a case of papillary adenocarcinoma occurring on ectopic thyroid tissue in the hyoid bone region. MATERIAL AND METHODS: A 57-year-old patient consulted for a cervical mass appearing 1 year before, with no signs of compression. The thyroid function was normal. RESULTS: The examination showed a firm tumefaction, fixed on the hyoid bone, with undetermined limits, measuring approximately 8 cm. The thyroid gland was in the normal pretracheal position. The cervical tomodensitometry objectified an osteolytic process centered on the hyoid bone with a fleshy zone of tissue density and calcifications. Cervical MRI provided a more precise assessment of the extension of the mass and confirmed the integrity of the other structures, in particular the endo-larynx. The tumor was totally resected via a cervical approach. The pathologic study suggested a papillary adenocarcinoma on ectopic thyroid tissue. The treatment was completed by a total thyroidectomy with normal histological results. CONCLUSIONS: Papillary adenocarcinoma on ectopic thyroid is a very rare situation. Its diagnosis is histological. Its treatment is primarily surgical, sometimes associated with radioiodine I-131 therapy. PMID- 19296928 TI - Vitamin C intake attenuates the degree of experimental atherosclerosis induced by periodontitis in the rat by decreasing oxidative stress. AB - OBJECTIVE: Periodontitis has been causally linked to cardiovascular disease, which is mediated through the oxidative stress induced by periodontitis. Since vitamin C has been suggested to limit oxidative damage, we hypothesized that vitamin C intake may reduce endothelial oxidative stress induced by periodontitis in the aorta. The aim of this study was to investigate the effects of vitamin C intake on the initiation of atherosclerosis in a ligature-induced rat periodontitis model. DESIGN: Eighteen 8-week-old-male Wistar rats were divided into three groups of six rats and all rats received daily fresh water and powdered food through out the 6-week study. In the vitamin C and periodontitis groups, periodontitis was ligature-induced for the first 4 weeks. In the vitamin C group, rats were given distilled water containing 1 g/L vitamin C for the 2 weeks after removing the ligature. RESULTS: In the periodontitis group, there was lipid deposition in the descending aorta and significant increases of serum level of hexanoyl-lysine (HEL), and aortic levels of nitrotyrosine expression, HEL expression and 8-hydroxydeoxyguanosine (8-OHdG) compared to the control group. Vitamin C intake significantly increased plasma vitamin C level and GSH:GSSG ratio (178% and 123%, respectively), and decreased level of serum HEL and aortic levels of nitrotyrosine, HEL and 8-OHdG (23%, 87%, 84%, and 38%, respectively). CONCLUSIONS: These results suggest that vitamin C intake attenuates the degree of experimental atherosclerosis induced by periodontitis in the rat by decreasing oxidative stress. PMID- 19296929 TI - Separating hoarding from OCD. AB - There is confusion about the status of excessive hoarding. In particular, there is an unresolved question about whether or not it is a manifestation of obsessive compulsive disorder (OCD). There are some definite similarities between 'compulsive' hoarding and OCD, but they are outweighed by the differences between them. It is proposed that the matter can be clarified by separating hoarding from OCD, and that this would advance clinical work and research. PMID- 19296930 TI - Early maturity of face recognition: no childhood development of holistic processing, novel face encoding, or face-space. AB - Historically, it was believed the perceptual mechanisms involved in individuating faces developed only very slowly over the course of childhood, and that adult levels of expertise were not reached until well into adolescence. Over the last 10 years, there has been some erosion of this view by demonstrations that all adult-like behavioural properties are qualitatively present in young children and infants. Determining the age of maturity, however, requires quantitative comparison across age groups, a task made difficult by the need to disentangle development in face perception from development in all the other cognitive factors that affect task performance. Here, we argue that full quantitative maturity is reached early, by 5-7 years at the latest and possibly earlier. This is based on a comprehensive literature review of results in the 5-years-to-adult age range, with particular focus on the results of the few previous studies that are methodologically suitable for quantitative comparison of face effects across age, plus three new experiments testing development of holistic/configural processing (faces versus objects, disproportionate inversion effect), ability to encode novel faces (assessed via implicit memory) and face-space (own-age bias). PMID- 19296931 TI - Seeing the world through another person's eyes: simulating selective attention via action observation. AB - Selective attention is usually considered an egocentric mechanism, biasing sensory information based on its behavioural relevance to oneself. This study provides evidence for an equivalent allocentric mechanism that allows passive observers to selectively attend to information from the perspective of another person. In a negative priming task, participants reached for a red target stimulus whilst ignoring a green distractor. Distractors located close to their hand were inhibited strongly, consistent with an egocentric frame of reference. When participants took turns with another person, the pattern of negative priming shifted to an allocentric frame of reference: locations close to the hand of the observed agent (but far away from the participant's hand) were inhibited strongly. This suggests that witnessing another's action leads the observer to simulate the same selective attention mechanisms such that they effectively perceive their surroundings from the other person's perspective. PMID- 19296932 TI - Somatosensory processing and body representation. AB - Recent years have seen increasing numbers of transcranial magnetic stimulation (TMS) studies focusing on somatosensory processing. Most have centered on the primary somatosensory functions of tactile detection, localization and discrimination, and have applied TMS to primary somatosensory areas. These studies confirm the basic functions of primary somatosensory areas, and the behavioural and physiological effects of different TMS protocols. Fewer studies, however, have investigated higher somatosensory function. Here, we review the somatosensory TMS literature both in and beyond primary somatosensory areas. We discuss the plausibility of modulating multisensory representations of one's own body via TMS, and highlight the potential for TMS to probe higher cognitive functions through the modulation of unimodal perceptual systems such as touch, vision or proprioception. PMID- 19296933 TI - VATER/VACTERL syndrome (vertebra/anus/cardiac/trachea/esophogus/radius/renal/limb anomalies) with a noncommunicating functioning uterine horn and a unicornuate uterus: a case report. AB - OBJECTIVE: To describe the presentation and outcome of a patient with an unusual association of VATER/VACTERL syndrome (vertebra/anus/cardiac/trachea/esophogus/radius/renal/limb) and noncommunicating functioning uterine horn and a unicornuate uterus. DESIGN: Descriptive case report. SETTING: A foundation trust in the United Kingdom. MATERIALS AND METHOD(S): A patient known to have VATER/VACTERL syndrome was diagnosed with a hematometra of a noncommunicating uterine horn and hematosalpinx. She underwent multidisciplinary surgery to remove the functioning uterine horn, after which she had a full recovery. CONCLUSION(S): This case documents the association of VATER/VACTERL syndrome with a unicornuate uterus and noncommunicating horn. PMID- 19296935 TI - Aberration of blastocyst microRNA expression is associated with human infertility. AB - OBJECTIVE: To examine human blastocyst microRNA (miRNA) expression in correlation with human infertility. MicroRNAs are small, noncoding RNA molecules that regulate gene expression via mechanisms such as degradation and translational suppression of targeted messenger RNAs. Recent data has pointed to the importance of miRNAs in disease states and during mouse embryo development. DESIGN: Descriptive study. SETTING: Nonprofit research foundation. PATIENT(S): Transferable quality human blastocysts donated with consent to research (n = 40). INTERVENTION(S): Quantitative real-time PCR. MAIN OUTCOME MEASURE(S): MicroRNA expression profile. RESULT(S): Morphologically similar blastocysts derived from patients with polycystic ovaries or male factor infertility exhibited a significant decrease in the expression of six miRNAs in comparison with donor fertile control blastocysts (P < 0.05). Annotation of predicted gene targets for these differentially expressed miRNAs included gene ontology (GO) biological processes involved in cell growth and maintenance and transcription as well as GO molecular functions implicated in nucleic acid binding and signal transducer activity. Three predicted miRNA target genes were selected for analysis and demonstrated significant altered expression consistent with aberrant miRNA profiles. CONCLUSION(S): This study describes for the first time that transferable quality blastocysts derived from infertile patients (male factor infertility and polycystic ovaries) possess aberrant miRNA profiles. With growing evidence indicating the importance of miRNAs during development, an association may exist with human infertility. PMID- 19296936 TI - Serologic markers of autoimmunity in women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate whether women with the polycystic ovary syndrome (PCOS) have an increased rate of elevated serum levels of common autoimmunologic parameters. DESIGN: Prospective case-control study. SETTING: Academic research institution. PATIENT(S): One hundred nine women with PCOS and 109 age-matched healthy controls. INTERVENTION(S): Peripheral venous puncture, ultrasonography, questionnaire. MAIN OUTCOME MEASURE(S): Serum levels of antinuclear antibodies (ANAs) were measured by immunometric enzyme immunoassay, serum levels of IgG class autoantibodies against histone, IgG class autoantibodies against nucleosomes, and IgG class autoantibodies against double-stranded (ds) DNA were measured by enzyme immunoassays. RESULT(S): Women with PCOS had significantly elevated serum levels of antihistone and anti-dsDNA antibodies, whereas serum levels of ANAs and antinucleosome antibodies were similar between the two groups. When serum levels of ANAs, antihistone, antinucleosomes, and anti-dsDNA antibodies were correlated with clinical and biochemical parameters, a significant correlation between serum levels of ANAs and serum TSH was established. CONCLUSION(S): Our study shows that serologic parameters of autoimmunity (i.e., antihistone and anti-dsDNA antibodies) are elevated in women with PCOS. A role of autoimmunologic processes in PCOS can be suspected. PMID- 19296937 TI - Kabuki syndrome and sex chromosomal anomalies: is it really an association? PMID- 19296938 TI - Setting up a medical abortion service for failure. PMID- 19296940 TI - Evidence of absence or absence of evidence? A reanalysis of the effects of low dose aspirin in in vitro fertilization. PMID- 19296941 TI - Embryo cryopreservation rescues cycles with premature luteinization. AB - OBJECTIVE: To determine whether embryo cryopreservation in cycles with elevated preovulatory P followed by thaw, extended culture, and transfer results in greater ongoing pregnancy rates than fresh blastocyst transfer. DESIGN: Retrospective matched cohort study. SETTING: Private fertility center. PATIENT(S): The study group consisted of 118 consecutive thaws of bipronucleate (2PN) oocytes derived from autologous cycles with elevated preovulatory P, resulting in 95 blastocyst transfers. The control group was selected by matching on the number of 2PN oocytes and patient age and consisted of 118 fresh cycles with elevated preovulatory P, including 108 fresh autologous blastocyst transfers. All patients were <41 years old at the time of stimulation. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Implantation and ongoing pregnancy rates. RESULT(S): The study group had significantly fewer blastocysts per 2PN oocyte than the control group (19.7% vs. 36.8%, respectively) and a significantly greater cancellation rate (19.5% vs. 8.5%, respectively). However, the ongoing pregnancy rate per cycle was significantly higher in the study group than in the control group (56.8% vs. 32.2%, respectively), resulting from greater rates of implantation (56.8% vs. 26.9%, respectively) and of ongoing pregnancy per transfer (70.5% vs. 35.2%, respectively). CONCLUSION(S): In cycles with elevated preovulatory P, the probabilities of implantation and ongoing pregnancy are increased if all 2PN oocytes are cryopreserved and subsequently thawed and cultured to the blastocyst stage before transfer. PMID- 19296942 TI - Glucose-regulated protein 78 (Grp78/BiP) is secreted by human oviduct epithelial cells and the recombinant protein modulates sperm-zona pellucida binding. AB - OBJECTIVE: To determine the secretion of Grp78 by human oviduct epithelial cells, its association to spermatozoa, and its involvement in gamete interaction. DESIGN: Prospective study. SETTING: Basic research laboratory. SUBJECT(S): Semen samples obtained from normozoospermic volunteers. Tubal tissue provided by patients undergoing hysterectomies. Oocytes collected from women undergoing IVF ET. INTERVENTION(S): Analysis of Grp78 expression and secretion by oviductal tissue. Gamete incubation with recombinant Grp78 (rec-Grp78). MAIN OUTCOME MEASURE(S): Assessment of protein expression and secretion by immunohistochemistry and Western immunoblotting, respectively. Evaluation of rec Grp78 binding to human spermatozoa by immunocytochemistry, and analysis of its effect upon gamete interaction using the hemizona assay. RESULT(S): Grp78 was found in the surface of oviduct epithelial cells. Soluble Grp78 was detected in oviductal fluids from women in the periovulatory period and in oviductal tissue conditioned medium. Rec-Grp78 was able to bind to the sperm acrosomal cap, and its presence during gamete interaction led to a decrease in the number of spermatozoa bound to the zona pellucida (ZP). When calcium ions from the incubation medium were replaced by strontium, rec-Grp78 enhanced sperm-ZP interaction. CONCLUSION(S): Grp78 is expressed and secreted by oviduct epithelial cells. The protein would bind to the gametes and may modulate their interaction in a calcium-dependent manner. PMID- 19296943 TI - Fertility and aging: do reproductive-aged Canadian women know what they need to know? AB - OBJECTIVE: Female fertility declines with age; however, women are increasingly delaying childbearing until later in their reproductive years. One of the factors that may contribute to this trend is a general lack of knowledge about the decline in fertility with age. DESIGN: Self-report survey. Questions pertained to participant demographics and childbearing intentions, and knowledge of the decline in fertility and increased risk of pregnancy loss with age. SETTING: The University of British Columbia in Vancouver, British Columbia, Canada. PATIENTS: Female undergraduate students (N = 360). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Knowledge of fertility over the life span, predictors of age of intended childbearing. RESULT(S): Although most women were aware that fertility declines with age, they significantly overestimated the chance of pregnancy at all ages and were not conscious of the steep rate of fertility decline. Surprisingly, women overestimated the chance of pregnancy loss at all ages, but did not generally identify a woman's age as the strongest risk factor for miscarriage. CONCLUSION(S): Education regarding the rate at which reproductive capacity declines with age is necessary to avoid unintended childlessness among female academics and professionals. PMID- 19296944 TI - Delayed delivery following unusual flare-up pelvic abscess after in vitro fertilization and embryo transfer. AB - OBJECTIVE: To report a case of delayed delivery of twins following pelvic abscess that required laparotomy after uncomplicated IVF-ET. DESIGN: Case report. SETTING: Department of Perinatology. PATIENT(S): A 32-year-old nulliparous woman after IVF-ET. INTERVENTION(S): In vitro fertilization and ET, laparotomy, observation in the hospital. MAIN OUTCOME MEASURE(S): Delayed delivery following pelvic abscess that required laparotomy after uncomplicated IVF-ET. RESULT(S): After a successful delayed delivery, the patient and her infant were discharged home. CONCLUSION(S): Pelvic abscess rarely complicates pregnancy. It cannot be excluded in women after IVF-ET. This report confirms the necessity to develop a comprehensive approach for continuation of complicated multiple pregnancies. PMID- 19296945 TI - Successful pregnancy and delivery of a healthy baby after endometrial biopsy treatment in an in vitro fertilization patient with severe Asherman syndrome. AB - OBJECTIVE: To implement the procedure of endometrial biopsy in a case of severe Asherman syndrome as a possible treatment to increase uterine receptivity. DESIGN: Case report. SETTING: IVF Unit, Kaplan Medical Center, Rehovot, Israel. PATIENT(S): A 29-year-old patient with severe Asherman syndrome, who underwent six operative hysteroscopies combined with hormonal treatment, and no functional receptive endometrium was achieved. INTERVENTION(S): We performed three endometrial biopsies on days 8, 12, and 21 of a progyluton-induced menstrual cycle, and a fourth biopsy on day 21 of the next induced menstrual cycle. After that cycle the patient underwent an IVF treatment. MAIN OUTCOME MEASURE(S): Ultrasound measurement of endometrial thickness, serum beta-hCG, sonography test for the presence of a gestational sac with heartbeat, and pregnancy follow-up until birth. RESULT(S): Biopsy treatment increased the thickness of the endometrium from unobservable by sonography to 7 mm on the day of hCG administration. The next IVF cycle resulted in implantation of an embryo and the birth of a healthy baby boy. CONCLUSION(S): Repeated endometrial biopsies may be used in patients with Asherman syndrome immediately after forming a uterine cavity by hysteroscopy to improve its receptivity. PMID- 19296946 TI - Anti-beta(2)-glycoprotein-I and anti-phosphatidylserine antibodies in women with spontaneous pregnancy loss. AB - OBJECTIVE: To evaluate the role of anti-beta(2)-glycoprotein-I (anti-beta(2)GPI ab) and anti-phosphatidylserine (aPS-ab) antibodies as a risk factor in both recurrent miscarriage (RM) and unexplained fetal losses (UFL). DESIGN: Retrospective, cohort study. SETTING: Vall d'Hebron University Hospital, Barcelona, Spain. PATIENT(S): 122 pregnant women divided in two groups: study group of 54 women with RM and/or UFL and control group of 68 pregnant without RM history. INTERVENTION(S): Analysis of lupus anticoagulant, anticardiolipin antibodies, and anti-beta(2)GP1 and aPS antibodies. MAIN OUTCOME MEASURE(S): Comparison of aPL antibody between groups. RESULT(S): The prevalence of aPL positive results was 8 out of 54 (14.8%) in the study group and 3 out of 68 (4.41%) in the controls. In the RM subgroup, the prevalence was 3 out of 25 (12%) versus 3 out of 68 (4.4%), and 7 out of 34 (20.6%) versus 3 out of 68 (4.4%) in UFL subgroup. As a whole, the prevalence of anti-beta(2)GP1-ab in the RM/UFL group showed a difference compared with controls but not aPS-ab. In the RM women, anti-beta(2)GP1-ab was positive in 3 out of 25 (12%) versus 1 out of 68 (1.5%) in controls and in 4 out of 34 versus 0 out of 68 cases in women with UFL. In the RM subgroup, aPS-ab was positive in 1 out of 25 (4%) versus 2 out of 68 (2.9%) in control group and in 3 out of 34 versus 2 out of 68 cases in women with UFL. CONCLUSION(S): Our results suggest that anti-beta(2)GP1-ab but not aPS-ab is related to RM/UFL and should be considered as a pregnancy-loss risk factor. PMID- 19296947 TI - Severe ureteral endometriosis: the intrinsic type is not so rare after complete surgical exeresis of deep endometriotic lesions. AB - OBJECTIVE: To evaluate the rate of intrinsic ureteral endometriosis in patients presenting with severe ureteral endometriosis. DESIGN: Observational study between June 1992 and December 2007. SETTING: University tertiary referral center. PATIENT(S): Twenty-nine patients presenting deeply infiltrating endometriosis (DIE) with severe ureteral endometriosis. Severe ureteral endometriosis was defined as DIE lesions causing significant obstruction to the urinary flow with ureteral stenosis. INTERVENTION(S): Complete surgical exeresis of DIE lesions. MAIN OUTCOME MEASURE(S): Pre- and peroperative evaluation associated with histologic analysis. Intrinsic ureteral endometriosis was defined as presence of DIE lesions infiltrating the ureteral muscularis. RESULT(S): In a series of 627 patients with histologic proved DIE, we observed 29 (4.6%) patients with severe ureteral endometriosis. Ureteral lesions (n = 34) were right sided in 7 (24.1%) patients, left sided in 17 (58.6%) patients, and bilateral in 5 (17.3%) patients. Eleven (37.9%) patients presented intrinsic lesions. Out of the 34 ureteral lesions 13 (38.2%) were intrinsic. In cases of radical ureteral surgery (n = 21 patients; n = 24 ureteral lesions) intrinsic ureteral DIE was observed in 52.4% (11 cases) of the patients and in 54.2% (13 cases) of the ureteral lesions. CONCLUSION(S): The prevalence of intrinsic ureteral endometriosis is underestimated. This result must be taken into account when specifying the surgical modalities for patients presenting with severe ureteral endometriosis. PMID- 19296948 TI - Immunoreactivity of progesterone receptor isoform B, nuclear factor kappaB, and IkappaBalpha in adenomyosis. AB - Compared with normal endometrium, progesterone receptor isoform B (PR-B) and IkappaBalpha immunoreactivity were statistically significantly reduced in ectopic as well as eutopic endometrium from women with adenomyosis while nuclear p65, p50, and p52 immunoreactivity were statistically significantly increased in ectopic and eutopic endometrium. Nuclear p65 immunoreactivity was positively associated with heavier menses, and decreased PR-B and increased nuclear p65 immunoreactivity in ectopic endometrium were statistically significantly associated with the severity of dysmenorrhea in women with adenomyosis. PMID- 19296949 TI - The effect of changing position and early ambulation after cardiac catheterization on patients' outcomes: a single-blind randomized controlled trial. AB - BACKGROUND: Cardiac catheterization is the gold standard diagnostic test for coronary heart diseases. In order to minimize the post-procedure complications, patients are restricted to prolonged bed rest that is always accompanied by fatigue and discomfort. OBJECTIVE: The aim of this study was to assess the effect of changing position and early ambulation on the level of comfort, satisfaction, and fatigue and on the amount of bleeding and hematoma after cardiac catheterization. PARTICIPANTS: A sample of 70 patients, who had undergone a non emergency 6-French cardiac catheterization via the femoral artery from September to November, 2006. METHODS: In a single-blind randomized controlled trial, each patient was randomly assigned to either the control or experimental group. The patients' position in the experimental group was intermittently changed during the first 6h after catheterization. Seven hours after the procedure, they were allowed to be ambulated and to undertake their self care activities. A pillow was placed under the patients' bodies. Patients in the control group were managed as routine; they were restricted to a 10-24h bed rest bed rest in supine position with the affected leg straight and immobilized and a sand bag on the puncture site for at least 8h. The levels of comfort, satisfaction and fatigue, and the amount of bleeding and hematoma were measured at regular intervals after the procedure. RESULTS: The patients in the experimental group had significantly higher comfort and satisfaction and lower fatigue levels than the control group at 3, 6, 8h and the next morning after catheterization (P<0.01). Changing patients' position according to the current protocol in the experimental group produced no significant increase in the amount of bleeding and hematoma when compared with the control group (P>0.05). CONCLUSION: The results of this study showed that the levels of comfort, satisfaction and fatigue after catheterization are related to the duration of bed rest and patients' position in bed. Changing patients' position accompanied by early ambulation after cardiac catheterization are associated with increasing comfort and satisfaction levels and decreasing the level of fatigue without increasing the amount of bleeding and hematoma. PMID- 19296950 TI - Compulsory community mental health treatment: literature review. AB - Following their introduction in the United States in the 1970s various forms of compulsory treatment in the community have been introduced internationally. Compulsory treatment in the community involves a statutory framework that mandates enforceable treatment in a community setting. Such frameworks can be categorized as preventative, least restrictive, or as having both preventative and least restrictive features. Research falls into two categories; descriptive, naturalistic studies and controlled and uncontrolled comparative studies. The research has produced equivocal results, and presents numerous methodological challenges. Where programmes have demonstrated improved outcomes debate continues as to whether these outcomes are associated with legal compulsion or enhanced service provision. Service user, family and clinician perspectives demonstrate a divergence of views within and across groups, with clinicians more strongly in support than service users. The issue of compulsory community treatment is an important one for nurses, who are often at the forefront of clinical service provision, in some cases in statutory roles. Critical reflection on the issue of compulsory community treatment requires understanding of the limitations of empirical investigations and of the various ethical and social policy issues involved. There is a need for further research into compulsory community treatment and possible alternatives. PMID- 19296951 TI - Carotid arterial elasticity is a sensitive atherosclerosis value reflecting visceral fat accumulation in obese subjects. AB - BACKGROUND: We previously reported the arterial elasticity value we measured to reflect the characteristic features of vessel walls, and to possibly be useful for detecting early stage atherosclerosis in type 2 diabetes. Obesity, especially visceral adiposity, is well known to play a crucial role in the development of metabolic disorders and atherosclerosis. To assess whether arterial elasticity value reflects the effect of obesity on atherosclerosis, we examined the associations of obesity characteristics with atherosclerosis values including arterial elasticity, carotid intima-media thickness (IMT) and pulse wave velocity (PWV). METHODS: Three atherosclerosis values were measured in 78 obese subjects (body mass index >/=30). We investigated the associations of atherosclerosis values with obesity-related parameters including abdominal fat accumulation determined by computed tomography. RESULTS: Arterial elasticity values were positively related to established atherosclerosis values, carotid IMT and PWV, in obese subjects. Age, systolic blood pressure and hypertension also correlated with these atherosclerosis values. Single regression analysis showed all three atherosclerosis values to correlate significantly with visceral fat area. Intriguingly, visceral fat area is an independent variable affecting arterial elasticity, but not IMT or PWV. Furthermore, multiple regression analysis revealed that arterial elasticity correlates strongly with visceral fat area. CONCLUSIONS: Arterial elasticity value we measure is a new parameter for evaluating atherosclerosis in subjects with visceral adiposity and more sensitive than the currently established atherosclerosis values, carotid IMT and PWV. Measuring arterial elasticity has the potential to reveal minute vascular changes, and may have broad clinical applications for evaluating early stage atherosclerosis. PMID- 19296952 TI - Low serum bilirubin concentration is associated with coronary artery calcification (CAC). AB - BACKGROUND: Bilirubin is a potent antioxidant and previous studies have reported the relationship between low serum bilirubin concentration and atherosclerosis. The purpose of this study was to assess the correlation between serum bilirubin concentration and coronary artery calcification (CAC). METHODS: This study consisted of 637 participants and we evaluated the relationship between CAC score determined by multislice computed tomography and serum bilirubin concentration. RESULTS: An inverse correlation was found between serum bilirubin concentration and log(CAC+1) (r=-0.361, P<0.0001). Multiple regression analysis also demonstrated that age (beta=0.261, P=0.0125), systolic blood pressure (beta=0.153, P=0.0237), uric acid (beta=0.126, P=0.0441), estimated glomerular filtration rate (beta=-0.139, P=0.0416) and serum bilirubin concentration (beta= 0.281, P<0.0001) were independent determinants of log(CAC+1). An increment of 1 micromol/L in serum bilirubin concentration was associated with 14% decrease in the odds for CAC score > or =400 after adjustment for several risk factors. Both age and SBP were also positively associated with CAC score > or =400, but the odds ratio for CAC score > or =400 was greater for every 1 micromol/L increment in serum bilirubin concentration than for every 1-year increment in age and 1 mmHg increment in SBP. CONCLUSIONS: Low serum bilirubin concentration is associated with coronary artery calcification. Serum bilirubin concentration can be measured easily in the clinical laboratory and applied in medical practice, and low serum bilirubin concentration would be useful as a provisional new risk factor of CAC. PMID- 19296954 TI - Investigation of the interaction mechanism of the recombinant human antibody MDJ8 and its fragments with chromatographic apatite phases. AB - The chromatographic behaviour of a recombinant human antibody (IgG(1)-subtype, kappa-light chain, MW: 149.5 kD, pI: 9.3) was investigated as a function of the buffer pH and buffer type (HEPES, phosphate, borate) on fluoroapatite and hydroxyapatite stationary phases. HEPES buffer was used at pH 7.0, phosphate buffer at pH 8.2 and borate buffer between pH 8.5 and 11. Elution was by a double gradient method of first a salt gradient from 0 to 1 M NaCl in the corresponding buffer, followed by a step gradient to 0.4 M sodium phosphate. Regardless of the pH and buffer type, the antibody eluted in the NaCl gradient; capacity factors decreased with increasing pH. At pH 11 the antibody eluted in the flow-through. Retention was thus dominated by electrostatic interaction throughout the investigated pH-range. Investigation of antibody fragments obtained by papain digestion (fc- and fab-fragments) and deglycosylated fc-fragments showed that the sugar structures had no influence on the chromatographic behaviour. Instead the chromatographic behaviour was dominated by that of the fab-fragment. zeta Potential measurements verified that the apatite surface bore a negative surface charge in the investigated pH range, while the antibody net surface charge switched from positive to negative as the pH increased. The corresponding isoionic point was a function of both the buffer concentration and the buffer species. However, above a pH of 8.3 the zeta-potential of the antibody generally was negative. Simulations of the molecular electrostatic potential of the antibody and the two fragments revealed the presence of a positively charged patch within the fab-fragment, which only disappeared above a pH of 10. Most likely this patch was responsible for the observed behaviour. PMID- 19296953 TI - Statin prevents plaque disruption in apoE-knockout mouse model through pleiotropic effect on acute inflammation. AB - Although it has been demonstrated that statins stabilize atherosclerotic lesions in animal models of advanced atherosclerosis, there is little evidence to suggest that statins have a preventive effect on plaque rupture itself. In the present study, we examined the effect of fluvastatin on plaque disruption using a simple and quick method of plaque disruption in carotid artery lesions in apolipoprotein E-deficient mice. Male apolipoprotein E-deficient mice received normal chow and underwent ligation of the left common carotid artery just proximal to its bifurcation. Four weeks later, a polyethylene cuff was placed around the artery immediately proximal to the ligation site. Fluvastatin (10mg/kg per day) was given by oral gavage every day starting at 3 days before cuff placement. The administration of fluvastatin suppressed atherosclerotic plaque disruption accompanied by luminal thrombi by 31.5% compared with controls at 4 days after the cuff was placed at the ligated carotid artery. Fluvastatin administration decreased matrix metalloproteinase-9 expression, gelatinolytic activity, endothelial adhesion molecules expression and neutrophil infiltration, and increased type I collagen content in the cuffed region. In summary, fluvastatin was found to prevent plaque disruption through pleiotropic effect on acute inflammation in an animal model using apolipoprotein E-deficient mice. PMID- 19296955 TI - Microsphere resin chromatography combined with microbial biotransformation for the separation and purification of salvianolic acid B in aqueous extract of roots of Salvia multiorrihza Bunge. AB - Salvianolic acid B was separated and purified from Salvia miltiorrhiza Bunge (danshen) by microbial transformation together with chromatography of microsphere resin. The aqueous extract of danshen was transformed by Fusarium graminearum in a bioreactor containing phosphate buffer (PBS), in which rosmarinic acid was transformed into danshensu and caffeic acid and the yield of salvianolic acid B was higher than 85%. After biotransformation, salvianolic acid B was purified by microsphere resin. A parallel test for making a comparison of microsphere resin chromatography between elution by methanol water solution and water was done. The purity of salvianolic acid B was up to 95% at the yield of 62% when impurities and salvianolic acid B were eluted by 45% and 55% methanol solution respectively. The purity of salvianolic acid B was up to 99% at the yield of 90% when distilled water was used to elute the impurities and salvianolic acid B. The total yield of salvianolic acid B was up to 75% at the purity over 99% while biotransformation combined with microsphere resin chromatography by water elution. Microbial biotransformation together with water elution of microsphere resin supplied an efficient method to eliminate the micromolecular impurities and a possible method to purify water-soluble compounds in traditional Chinese medicine. PMID- 19296957 TI - Multidimensional gas chromatography-mass spectrometry determination of 3-alkyl-2 methoxypyrazines in wine and must. A comparison of solid-phase extraction and headspace solid-phase extraction methods. AB - Two different strategies for the quantitative determination of 3-alkyl-2 methoxypyrazines in wine and must have been developed and validated. Comparison between both the techniques has been presented and the most adequate has been applied to the determination of these compounds in different samples of wine, made from several varieties of grapes, and also in different samples of must made from Cabernet Sauvignon grapes. Both the methods consisted of a dynamic headspace coupled with a solid-phase extraction (HS-SPE) and solid-phase extraction (SPE) directly from the sample, coupled with multidimensional gas chromatography-mass spectrometry system (MDGC-MS). Both of them require resins LiChrolut EN, and analyte elution has been carried out with dichloromethane. The repeatability of both methodologies was evaluated at two concentration levels. The relative standard deviations (RSD%) were acceptable in every case, but smaller when working with HS-SPE. The recoveries obtained for the three analytes with the two methodologies were almost 100%, with the exception of IBMP, which had a recovery of only 70% with HS-SPE. The linearity was satisfactory with both methods for the range of occurrence of methoxypyrazines in wine and must. The limits of detection of the direct SPE technique were much lower than those of HS-SPE in every case. Direct SPE method detection limits ranged from 0.09 to 0.15 ng L(-1). The method based on direct SPE was chosen finally because it had better detection limits and was easier and quicker than the HS-SPE-based method. It has been applied to the determination of these components in 36 wine and 17 musts samples. The quantitative results suggest that the Spanish wines show meaningless amounts of these compounds. IBMP has been found just in between 1.9 and 15 ng L(-1). PMID- 19296956 TI - Determination of cyclopiazonic acid in food and feeds by liquid chromatography tandem mass spectrometry. AB - A new, fast and efficient multiple reaction monitoring (MRM) high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of cyclopiazonic acid (CPA) in mixed feed, wheat, peanuts and rice is presented. The analytical methodology involves sample extraction with an alkaline methanol-water mixture, defatting with hexane and quantification using HPLC-MS/MS without further treatment of sample extracts. Reversed-phase liquid chromatography using a C18 stationary phase coupled to negative mode electrospray triple quadrupole tandem mass spectrometry was applied. The limit of detection was 5 microg/kg while the limit of quantification was 20 microg/kg in the matrices investigated. The detector response was found to be linear over the range 25-250 microg/kg in feed and 25-500 microg/kg in wheat, peanuts and rice. The mean overall recoveries (n=18) of CPA varied from 79% to 114% in the range of concentrations studied over a period of 4 months. Mean recoveries (n=3 or 6) of CPA in wheat, peanuts and rice varied from 70% to 111%, 77% to 116% and 69% to 92%, respectively. The method was successfully applied to the analysis of feed and rice samples artificially infected with the fungal strain Penicillium commune, where the toxin was found at different levels. PMID- 19296958 TI - Isolation of xanthyletin, an inhibitor of ants' symbiotic fungus, by high-speed counter-current chromatography. AB - Xanthyletin, an inhibitor of symbiotic fungus (Leucoagaricus gongylophorus) of leaf-cutting ant (Atta sexdens rubropilosa), as well as suberosin, seselin and xanthoxyletin were isolated from Citrus sinensis grafted on Citrus limonia. A two phase solvent system composed of hexane/ethanol/acetonitrile/water (10:8:1:1, v/v) was used for the high-speed counter-current chromatographic isolation of xanthyletin with high yield and over 99% purity as determined by liquid and gas chromatography with mass spectrometry detection. Identifications were performed by UV spectra, IR spectra, (1)H NMR and (13)C NMR. PMID- 19296959 TI - Poly(1-allylimidazole)-grafted silica, a new specific stationary phase for reversed-phase and anion-exchange liquid chromatography. AB - A new specific stationary phase based on poly(1-allylimidazole)-grafted silica has been synthesized and characterized, by infrared spectra, elemental analysis, thermogravimetric analysis and X-ray photoelectron spectroscopy. The results of test showed that poly(1-allylimidazole) can effectively mask the residual silanol groups and reduce the adverse effect of residual silanol. Using this stationary phase, phenol compounds, aniline compounds, and polycyclic aromatic hydrocarbons were successfully separated with symmetric peak shapes in the reversed-phase chromatography. Inorganic anions (IO(3)(-), BrO(3)(-), Br(-), NO(3)(-), I(-), SCN(-)) were also separated completely in the anion-exchange chromatography using sodium chloride solution as the mobile phase. The effects of pH and the concentration of eluent on the separation of inorganic anions were studied. The separation mechanism appears to involve the mixed interactions of hydrogen bonding, hydrophobic, pi-pi, electrostatic, and anion-exchange interactions. PMID- 19296960 TI - Optimization of matrix solid-phase dispersion extraction method for the analysis of isoflavones in Trifolium pratense. AB - A method based on matrix solid-phase dispersion (MSPD) has been developed for the determination of 12 isoflavones in Trifolium pratense L. Dried leaf samples were blended with C(18), placed in small columns and isoflavones extracted with dichloromethane-methanol. Analyses were performed by high performance liquid chromatography with diode array detection (HPLC-DAD) with 2-methoxyflavone as internal standard. Several dispersants, eluents and clean-up steps were tested during the optimization of the process in order to obtain the best selectivity and yields. Mean recoveries ranged from 70% to 119%, with relative standard deviations <18%. The limits of detection were between 0.006 mg/l for biochanin A and 0.108 mg/l for daidzin. The performance of the optimized method in real samples was compared with a conventional method based in solid-liquid extraction (SLE). PMID- 19296961 TI - Determination of alkenes in cracking products by normal-phase high-performance liquid chromatography-diode array detection. AB - Alkene content determinations in fluid catalytic cracking (FCC) liquid products were performed by means of normal-phase high-performance liquid chromatography (NP-HPLC) with diode array detection (UV/DAD). Separation of alkenes from aromatic hydrocarbons was performed on amino-modified silica gel column with n heptane as mobile phase. The column has a little affinity to alkenes and saturated hydrocarbons and a pronounced affinity to aromatic compounds. The problem of alkenes and saturates co-elution on this column type was overcome with the detection system, UV/DAD, sensitive and selective to alkenes, while saturates are inactive in UV field. Total alkene content was determined as a sum of mono- and dialkene groups quantified by external standard method. Validation and verification of the developed method proved their applicability. The following criteria were used to validate the HPLC-DAD method: selectivity, linearity, precision, limits of detection and quantification. Alkene contents were quantified with the external standard method of wide calibration range, so both low and high alkene contents can be determined by the single calibration. Correlation coefficients were higher than 0.99. Precision was evaluated as repeatability and intermediary precision with relative standard deviations less than 5%. Some structural investigation of alkene groups was performed to confirm the assumption. Proposed method was compared with certified NMR method. Six commercial motor gasoline samples were analyzed by these two methods. Obtained results indicate good agreement between alkene content determined by both methods. The developed method was applied to the determination of alkene content in liquid FCC products in the boiling range from 70 degrees C to 190 degrees C. PMID- 19296962 TI - The effect of guest molecular architecture and host crystallinity upon the mechanism of the intercalation reaction. AB - The intercalation process of alpha-zirconium phosphate (alpha-ZrP) was investigated by using two alpha-ZrP samples with different levels of crystallinity and two structurally different intercalating molecules, i.e., linear hexylamine and non-planar cyclohexylamine. The results show that the intercalation energy barrier, which is affected by both host alpha-ZrP and guest intercalating molecules, has a significant effect on the intercalation process. When the intercalation energy barrier is relatively low, the interlayer distance of alpha-ZrP expands continuously with increasing amount of intercalating molecules. When the energy barrier reaches a certain level, the interlayer distance expansion becomes stepwise. The observed differences in the intercalation process correspond well with the geometric arrangement of the intercalated molecules inside the gallery of alpha-ZrP. PMID- 19296963 TI - A single-component photo-responsive fluid based on a gemini surfactant with an azobenzene spacer. AB - A new gemini surfactant with an azobenzene group incorporated in the spacer has been synthesized. In water, this new compound forms a photo-responsive fluid, and the properties have been characterized by rheology and freeze-fracture electron microscopy (FF-TEM). UV/Vis irradiation causes a reversible change in zero shear viscosity of about 5 orders of magnitude. The different rheological properties correspond to different aggregation states in solution. A densely entangled network and small discrete micelles are respectively responsible for the high viscosity and the low viscosity state, as revealed by FF-TEM. The simplicity and high photo-efficiency of this single-component system act as a foundation for designing novel stimuli-responsive materials. PMID- 19296964 TI - Ultrasensitive detection of mercury (II) ions using electrochemical surface plasmon resonance with magnetohydrodynamic convection. AB - A high-precision technique that can detect mercury (II) ions down to 1 fM concentration in aqueous solutions is introduced. The technique combines the conventional electrochemical method, surface plasmon resonance (SPR), and magnetohydrodynamic (MHD) convection. Mercury ions are electroplated onto a gold SPR sensing surface, and then detected quantitatively by applying a potential scan with cyclic voltammetry. Both the SPR angular shift and the electrochemical current signal are recorded for identification and quantification of the mercury ions. The detection sensitivity is further enhanced by applying an MHD convection in the presence of a magnetic field, which does not require any moving parts intruding into the aqueous solution. The technique thus has a great advantage for small detection volume. In the presence of supporting electrolytes, 1 mM nitric acid and 10 mM potassium nitrate, Hg(2+) ionic solutions with concentrations ranging from 1 fM to 1 microM are tested under different magnetic flux densities of B=0, 0.27, 0.53, and 0.71 T. The experimental results demonstrate that the stripping signals of the 1 fM to 1 microM Hg(2+) ions are enhanced by 10-60% with the flux density B=0.71 T. PMID- 19296965 TI - Individual differences in trajectories of arithmetical development in typically achieving 5- to 7-year-olds. AB - The arithmetical performance of typically achieving 5- to 7-year-olds (N=29) was measured at four 6-month intervals. The same seven tasks were used at each time point: exact calculation, story problems, approximate arithmetic, place value, calculation principles, forced retrieval, and written problems. Although group analysis showed mostly linear growth over the 18-month period, analysis of individual differences revealed a much more complex picture. Some children exhibited marked variation in performance across the seven tasks, including evidence of difficulty in some cases. Individual growth patterns also showed differences in developmental trajectories between children on each task and within children across tasks. The findings support the idea of the componential nature of arithmetical ability and underscore the need for further longitudinal research on typically achieving children and of careful consideration of individual differences. PMID- 19296966 TI - Dendritic pathology in Alzheimer's disease. AB - Dendritic pathology and decrease of dendritic spine density are prominent phenomena in early cases of Alzheimer's disease, which correlate significantly with the progressive decline of the mental faculties. In previous studies we have described the pathological alterations of the dendrites and the dendritic spines in the prefrontal area of the cortex and the cerebellum. In this study we attempted to describe the morphological alterations of the dendrites and the dendritic spines, quantifying them in the acoustic and the visual cortices of eleven cases of Alzheimer's disease, applying Golgi staining and electron microscopy. In addition, describing also the ultrastructural changes of the mitochondria in the dendritic profiles and the dendritic spines we noticed that mitochondrial pathology correlates substantially with the dystrophic dendrites, the loss of dendritic branches and the pathological alteration of the dendritic spines. We would hypothesize that mitochondrial alterations may play a very important role in dendritic degeneration and the loss of dendritic spines and we should have thought that therapeutic strategies protecting the mitochondria may be beneficial in Alzheimer's disease. PMID- 19296967 TI - The adjustable continence therapy system for recurrent female stress urinary incontinence: 1-year results of the North America Clinical Study Group. AB - PURPOSE: We determined the efficacy, safety, adjustability and technical feasibility of the adjustable continence therapy device (Uromedica, Plymouth, Minnesota) for the treatment of recurrent female stress urinary incontinence. MATERIALS AND METHODS: Female patients with recurrent stress urinary incontinence were enrolled in the study and a defined set of exclusionary criteria were followed. Baseline and regular followup tests to determine eligibility, and to measure subjective and objective improvement were performed. A trocar was passed fluoroscopically and with digital vaginal guidance to the urethrovesical junction through small incisions between the labia majora and minora. The adjustable continence therapy device was delivered and the balloons were filled with isotonic contrast. The injection ports for balloon inflation were placed in a subcutaneous pocket in each labia majora. Device adjustments were performed percutaneously in the clinic postoperatively. An approved investigational device exemption Food and Drug Administration protocol was followed to record all adverse events. RESULTS: A total of 162 subjects underwent implantation with 1 year of data available on 140. Mean Stamey score improved by 1 grade or more in 76.4% (107 of 140) of subjects. Improvement in the mean incontinence quality of life questionnaire score was noted at 36.5 to 70.7 (p <0.001). Reductions in mean Urogenital Distress Inventory (60.3 to 33.4) and Incontinence Impact Questionnaire (54.4 to 23.4) scores also occurred (p <0.001). Mean provocative pad weight decreased from 49.6 to 11.2 gm (p <0.001). Of the patients 52% (67 of 130) were dry at 1 year (less than 2 gm on provocative pad weight testing) and 80% (102 of 126) were improved (greater than 50% reduction on provocative pad weight testing). Complications occurred in 24.4% (38 of 156) of patients. Explantation was required in 18.3% (28 of 153) of the patients during 1 year. In terms of the complications 96.0% were considered to be mild or moderate. CONCLUSIONS: The Uromedica adjustable continence therapy device is an effective, simple, safe and minimally invasive treatment for recurrent female stress urinary incontinence. It can be easily adjusted percutaneously to enhance efficacy and complications are usually easily manageable. Explantation does not preclude later repeat implantation. PMID- 19296968 TI - Pediatric urinary stone disease--does age matter? AB - PURPOSE: It has been proposed that younger children are less likely to pass renal calculi spontaneously, and that children younger than 10 years are more likely to have an identifiable metabolic abnormality and subsequently a higher risk of recurrence. We report our clinical outcomes in children with urinary calculi, specifically examining these factors. MATERIALS AND METHODS: We performed a retrospective review of all pediatric patients diagnosed with renal or ureteral calculi at our institution between 2000 and 2007. Of 150 patients evaluated and treated during this period 80 (86 stones) had sufficient followup data to be included. Patients were divided into 2 groups according to age, namely 10 years or younger and older than 10 years. There were 39 patients in the younger group and 41 patients in the older group. Stone size and location, successful passage or intervention, recurrence and 24-hour urine metabolic study results were recorded. RESULTS: Of the younger cohort stones were ureteral in 43% and renal in 57%. The opposite trend was seen in older patients, with 69% having ureteral and 31% having renal stones (p = 0.02). Mean stone size (greatest dimension) did not differ significantly between the older and younger groups (6.9 mm vs 5.5 mm, p = 0.17). Overall stone passage rate was 34% for younger and 29% for older patients (p = 0.65). No significant mean size differences in passed stones existed between the groups (3.2 mm vs 2.5 mm, p = 0.31). Overall younger vs older ureteral stone passage rate was 37% vs 41% (p = 0.58), and for renal stones it was 32% vs 0%. Stones recurred in 7 younger and 6 older patients. CONCLUSIONS: Younger children were more likely to present with renal stones, while older children had more ureteral stones. Overall children 10 years old or younger are as likely to pass stones as older children. Renal stones are more likely to be successfully managed expectantly in younger children. Metabolic abnormalities and stone recurrences are observed at similar rates between younger and older children. PMID- 19296971 TI - Assessing the surgical decision making abilities of novice and proficient urologists. AB - PURPOSE: We examined whether the Surgical Decision Making Rating Scale can measure a difference in surgical judgment among urologists at various levels of training. MATERIALS AND METHODS: A total of 25 medical students, urology residents and staff urologists viewed clips from 8 select urological procedures and verbalized their thought processes. The clips were ordered in increasing complexity from lower level tasks (catheterization and cystoscopy) to more advanced procedures (laser lithotripsy, and open and laparoscopic prostatectomy and nephrectomy). Performance was transcribed and blindly rated using the previously validated rating scale. Subjects were also asked to self-evaluate their performance using this scale. RESULTS: Overall the rating scale distinguished the training level across knowledge domains (anatomy and management of the current task) and judgment domains (avoiding complications, higher reasoning and immediate surgical planning). The mean score across all training levels was 112 of 200 (range 51 to 161). Scale performance showed a significant correlation with seniority (rho = 0.96, p <0.05). This trend persisted when performance was analyzed separately for knowledge and judgment domain elements (rho = 0.95 and 0.96, respectively, each p <0.05). Self-evaluation correlated well with blinded evaluation across all levels of training (rho = 0.87, p = 0.01). CONCLUSIONS: The Surgical Decision Making Rating Scale can reliably detect differences in knowledge and surgical judgment among medical students, urology residents and staff urologists. This tool has potential applications for evaluating trainees and determining subjects with proficient decision making abilities. It also shows a significant correlation between self-rated performance and blinded evaluation. PMID- 19296969 TI - Complications in women undergoing Burch colposuspension versus autologous rectus fascial sling for stress urinary incontinence. AB - PURPOSE: We determined the clinicodemographic factors associated with complications of continence procedures, the impact of concomitant surgery on the complication rate and the relationship between the incidence of cystitis and the method of postoperative bladder drainage. MATERIALS AND METHODS: We reviewed serious adverse events and adverse events in the Stress Incontinence Surgical Efficacy Trial, a randomized trial comparing Burch colposuspension to the autologous rectus fascial sling. Clinicodemographic variables were analyzed to determine those associated with adverse events using logistic regression analysis. Complications were stratified based on the presence or absence of concomitant surgery. Differences in complication rates (controlling for concomitant surgery) and cystitis rates (controlling for the bladder emptying method) were compared using Fisher's exact test. RESULTS: Blood loss (p = 0.0002) and operative time (p <0.0001) were significantly associated with an adverse event. Patients who underwent concomitant surgery had a significantly higher serious adverse event rate (14.2% vs 7.3%, p = 0.01) and adverse event rate (60.5% vs 48%, p <0.01) than patients who underwent continence surgery alone. Cystitis rates were higher in the sling vs the Burch group up to 6 weeks postoperatively regardless of concomitant surgery status (p <0.01). Intermittent self-catheterization increased the cystitis rate by 17% and 23% in the Burch and sling groups, respectively. CONCLUSIONS: Concomitant surgery at continence surgery increased the risk of complications. Sling surgery was associated with a higher risk of cystitis within the first 6 weeks postoperatively. Intermittent self-catheterization increased the risk of cystitis in each group. Complications were associated with surgical factors and not with patient related factors. PMID- 19296973 TI - Surgical resection for suburethral sling complications after treatment for stress urinary incontinence. AB - PURPOSE: Suburethral tapes have been widely adopted to treat stress urinary incontinence. Further resection of such tapes may be necessary in certain cases. We review our experience and assess urinary functional outcomes. MATERIALS AND METHODS: We retrospectively reviewed the data on all women referred to our institution between 2001 and 2007 for suburethral tape related complications and on those who had the tape surgically removed. Complete or partial resection was achieved after assessment, including endoscopic and urodynamic assessment. RESULTS: A total of 75 women with a mean age of 60.7 years (range 28 to 78) were included in the study. The tape used was transvaginal in 58 cases (77.3%) and transobturator in 17 (22.7%). There were different complications, such as erosion in 16% of cases, vaginal extrusion in 24%, bladder outlet obstruction in 45%, chronic pelvic pain in 21%, and de novo urinary incontinence and urgency in 12%. Resection was done a mean +/- SD of 33 +/- 22 months (range 6 to 80) after tape placement. Of the 58 women with transvaginal tape the tape was completely removed by laparoscopy in 30 (51%). Four of the 17 transobturator slings (23%) were completely removed by laparoscopy (1) and via a low gynecological approach (3). The remaining slings were partially resected via a gynecological approach. At a mean followup of 38.4 months (range 12 to 72) incontinence recurred in 39 women (52%) after partial (18) and complete (21) resection. CONCLUSIONS: In rare women who experience crippling symptoms after suburethral sling implantation urologists must be aware that the decision to completely or partially resect the tape can help resolve symptoms. PMID- 19296974 TI - Benign prostatic obstruction and parkinson's disease--should transurethral resection of the prostate be avoided? AB - PURPOSE: According to the literature transurethral resection of the prostate in patients with Parkinson's disease has an increased risk of postoperative urinary incontinence. However, this conclusion might have been reached because some patients with multiple system atrophy incorrectly diagnosed as Parkinson's disease were included in these reports. Therefore, we investigated the outcome of transurethral prostate resection in patients with a secure neurological diagnosis of Parkinson's disease. MATERIALS AND METHODS: A total of 23 patients with Parkinson's disease who underwent transurethral prostate resection for benign prostatic obstruction were evaluated retrospectively. Subsequent neurological developments in patients were followed, ensuring that those with multiple system atrophy had not been included in analysis. RESULTS: At transurethral prostate resection median patient age was 73 years, median duration of Parkinson's disease before the resection was 3 years, and median Hoehn and Yahr scale was 2. Of the 14 patients with a preoperative indwelling urinary catheter transurethral prostate resection restored voiding in 9 (64%) and only 5 (36%) required catheterization postoperatively. Of the 10 patients with preoperative urge urinary incontinence, continence was restored in 5 and improved in 3 following transurethral prostate resection. There were no cases of de novo urinary incontinence after transurethral prostate resection. At a median postoperative followup of 3 years transurethral prostate resection was successful in 16 of the 23 patients (70%). CONCLUSIONS: Transurethral prostate resection for benign prostatic obstruction in patients with Parkinson's disease may be successful in up to 70% and the risk of de novo urinary incontinence seems minimal. Thus, Parkinson's disease should no longer be considered a contraindication for transurethral prostate resection provided that preoperative investigations including urodynamic assessment indicate prostatic bladder outlet obstruction. PMID- 19296975 TI - Women overestimate daytime urinary frequency: the importance of the bladder diary. AB - PURPOSE: We determined whether women with urinary incontinence reliably report urinary frequency. MATERIALS AND METHODS: We retrospectively reviewed the medical records of 1,136 women who underwent a suburethral sling operation from 1999 to 2007. A routine form was used to document medical history, urinary symptoms, physical examination and urodynamics. A total of 601 patients with a mean +/- SD age of 59.2 +/- 11.6 years (range 30 to 91) who completed a bladder diary were included in this study. Patients were categorized into 4 levels of urinary frequency, and medical histories and bladder diaries were compared. The Spearman correlation coefficient was used to determine the degree of agreement between histories and diaries. RESULTS: According to the diaries 535 patients (89%) voided 4 to 10 times during the day and 527 (87.6%) voided no more than twice at night. Only 47% of the women were accurate about daytime frequency. On the other hand, 93% of the women were accurate about the nighttime estimation. Of the women 51% overestimated daytime frequency. Overestimation was marked, especially in patients who reported a daytime frequency of greater than 10 voids per day. There was weak correlation between daytime urinary frequency obtained from medical histories and the frequency recorded in diaries (r = 0.31, p = 0.011). However, nighttime urinary frequencies correlated highly (r = 0.79, p = 0.02). Overestimation rates were similar in patients with and without overactive bladder. CONCLUSIONS: Our results suggest that half of patients tend to overestimate daytime urinary frequency. However, the accuracy rate of nighttime frequency is much higher. A bladder diary provides invaluable information about urinary frequency and it should be an integral part of the routine assessment in women with lower urinary tract symptoms. PMID- 19296976 TI - The bone anchor suburethral synthetic sling for iatrogenic male incontinence: critical evaluation at a mean 3-year followup. AB - PURPOSE: We retrospectively report objective and subjective outcomes in 40 male patients who underwent bone anchored suburethral synthetic sling positioning for stress urinary incontinence due to intrinsic sphincter deficiency. MATERIALS AND METHODS: Patients with stress urinary incontinence due to radical retropubic prostatectomy (32), robot assisted laparoscopic prostatectomy (3) and transurethral prostate resection (5) underwent bone anchored suburethral synthetic sling positioning between December 2002 and December 2007. Mean followup was 35.2 months (range 2 to 62). Previous anti-incontinence procedures, radiotherapy and transurethral procedures due to urethral stricture were performed in 5, 11 and 5 patients, respectively. Before and after surgery patients were evaluated by physical examination, urethral cystoscopy, urodynamics, a 1-hour pad test and a quality of life questionnaire. Patients were stratified into 3 groups, including group 1-cured (dry with a pad weight of 0 to 1 gm), group 2-improved (mild to moderate incontinence with a pad weight of 2 to 50 gm) and group 3-failed (patient condition unchanged with a pad weight of greater than 50 gm). RESULTS: At the final followup visit 22 (55%), 5 (12.5%) and 13 patients (32.5%) were cured, improved and failed, respectively. Mean pad weight significantly decreased to 51.3 gm in 54% of cases, while the mean total questionnaire score significantly increased to 72.9 in 65% and abdominal leak point pressure significantly increased to 92.5 cm H(2)O in 52%. Statistical analysis showed a significant association between preoperative radiotherapy and treatment failure (85% of patients). Complications were perineal pain in 73% of cases, detrusor overactivity in 5% and sling infection in 15%. CONCLUSIONS: The bone anchored suburethral synthetic sling is a simple and attractive procedure that can produce immediate good results with low morbidity, especially when strictly selected patients are treated. Radiotherapy remains a strong predictor of failure. PMID- 19296977 TI - Contemporary surgical management of upper urinary tract calculi. AB - PURPOSE: Upper urinary tract calculi are treated with multiple technologies including shock wave lithotripsy, ureteroscopy and percutaneous nephrolithotomy. Our knowledge of surgical practice patterns in the treatment of these calculi is limited. We performed a study of the surgical practice logs submitted to the American Board of Urology by candidates for initial certification and recertification to characterize the manner in which renal and ureteral calculi are treated. MATERIALS AND METHODS: Logs from initial certification, first recertification and second recertification cohorts were reviewed. CPT codes were used as search criteria, and included 50590 (shock wave lithotripsy), 52352 (ureteroscopy, stone removal), 52353 (ureteroscopy, lithotripsy), 50080 (percutaneous nephrolithotomy for stones less than 2 cm) and 50081 (percutaneous nephrolithotomy for stones greater than 2 cm). RESULTS: For the initial certification cohort surgical logs from 2004 to 2008 were reviewed and 1,065 individuals were identified. For the 2 recertification cohorts logs from 2003 to 2007 were reviewed, with 1,120 individuals identified in the first recertification cohort, and 831 identified in the second recertification cohort. Candidates for initial certification used ureteroscopy in the majority of stone removal procedures (52.0%), and candidates for first and second recertification used shock wave lithotripsy in the majority of their procedures (57.4% and 60.5%, respectively). There was a decreasing use of percutaneous nephrolithotomy across the cohorts with 6.8% in the initial, 4.5% in the first and 2.6% in the second recertification cohort. CONCLUSIONS: Provider specific attributes may affect how upper tract calculi are treated. Urologists in the initial certification cohort claimed the greatest use of endoscopic treatment modalities and most commonly performed ureteroscopy. Shock wave lithotripsy was more commonly used by the 2 recertification cohorts, comprised of more senior urologists. PMID- 19296978 TI - The safety and efficacy of optical urethrotomy using a spongiosum block with sedation: a comparative nonrandomized study. AB - PURPOSE: Optical urethrotomy is generally performed with the patient under general or major regional anesthesia. We determined the safety and efficacy of optical urethrotomy using a spongiosum block with sedation for anterior urethral stricture in a comparative, nonrandomized study. MATERIALS AND METHODS: In 32 patients with anterior urethral stricture optical urethrotomy was performed under general/major regional anesthesia in 16 patients (group 1) or a spongiosum block and sedation in 16 (group 2). In group 2 a total of 2 to 3 ml 1% lidocaine were slowly injected into the glans penis. Standard optical urethrotomy was performed immediately with a cold cut knife. RESULTS: The 2 groups were matching in terms of patient age, and stricture cause and length. Optical urethrotomy was successfully completed in all patients in group 1 and in 15 of 16 in group 2. In group 2, 15 patients (94%) had no pain or discomfort. One patient reported moderate discomfort and the procedure was abandoned. In group 2 none of the patients required parental analgesia post procedure. The first year recurrence was not significantly different in the 2 groups (p = 0.192). The anesthetic effect lasted for about an hour and was satisfactory without any complications. Pain score on the visual analogue scale was not different in the 2 groups. CONCLUSIONS: Optical urethrotomy using a spongiosum block with sedation is as safe and effective as using regional or general anesthesia, particularly in patients who are more ill. The shorter operative time in the local anesthesia group could also make it cost-effective. PMID- 19296979 TI - Oral contraceptive use and incident urinary incontinence in premenopausal women. AB - PURPOSE: Recent data in postmenopausal women indicate that current and past use of exogenous hormones is related to urinary incontinence risk. Little is known about exogenous hormones and risk of urinary incontinence in younger women. We investigated the association between oral contraceptive pills and incident urinary incontinence in premenopausal women enrolled in the Nurses' Health Study II. MATERIALS AND METHODS: Participants reported use of oral contraceptive pills from 1989 to 2001. Among 21,864 premenopausal women 37 to 54 years old reporting no urinary incontinence in 2001 we identified 749 with incident urinary incontinence at least weekly between 2001 and 2003. Odds ratios and 95% confidence intervals were estimated using multivariable logistic regression. RESULTS: Women who had ever used oral contraceptive pills had a statistically significant 27% (95% CI 1-59) increased odds of experiencing urinary incontinence at least weekly compared with those who never used oral contraceptive pills. In women with 10 or more years of use the odds ratio increased to 1.48 (95% CI 1.13 1.95). Ever using oral contraceptive pills was specifically associated with urgency urinary incontinence (OR 2.48, 95% CI 1.07-5.76) rather than stress urinary incontinence (OR 1.04, 95% CI 0.78-1.40). Although we had limited information on urinary tract infection, control for urinary tract infection did not alter these findings. CONCLUSIONS: Use of oral contraceptive pills may be associated with a modest increase in the odds of urinary incontinence among premenopausal women. However, this is one of the first reports of such an association and, thus, further research is needed to confirm our findings and investigate possible mechanisms. PMID- 19296980 TI - Sexual function among overweight and obese women with urinary incontinence in a randomized controlled trial of an intensive behavioral weight loss intervention. AB - PURPOSE: We examined sexual function in overweight and obese women with urinary incontinence, and evaluated the effects of an intensive behavioral weight reduction intervention on sexual function in this population. MATERIALS AND METHODS: A total of 338 overweight and obese women reporting 10 or more incontinence episodes weekly were randomized to an intensive behavioral change (226) or structured education program (112) for 6 months. Sexual function was assessed using self-administered questionnaires. Multivariate regression was used to examine factors associated with baseline and 6-month change in sexual function as well as intervention effects. RESULTS: Two-thirds of participants (233) were sexually active at baseline but more than half (188) reported low desire and a quarter (91) were sexually dissatisfied. More than half of sexually active participants (123) reported problems with arousal, lubrication, orgasm or incontinence during sex. Compared to controls women in the intervention group demonstrated a borderline increase in frequency of sexual activity at 6 months (OR 1.34, 95% CI 0.99-1.81, p = 0.06) but no differences in satisfaction (OR 1.28, 95% CI 0.83-1.99, p = 0.26), desire (OR 1.12, 95% CI 0.79-1.61, p = 0.52) or problems (beta +/- SE 0.03 +/- 0.07, p = 0.68 for intervention effects on problems score). Neither clinical incontinence severity nor body mass index was independently associated with baseline or 6-month change in function (p >0.10 for all). CONCLUSIONS: Sexual dysfunction is common in overweight and obese women with incontinence but the severity of this dysfunction may not be directly related to the severity of incontinence or obesity. An intensive 6-month behavioral weight reduction intervention did not significantly improve sexual function in this population relative to controls. PMID- 19296981 TI - Patients with primary hyperparathyroidism--why do some form stones? AB - PURPOSE: Primary hyperparathyroidism is a common cause of hypercalcemia with resulting hypercalciuria and a subset of these patients with primary hyperparathyroidism present with urolithiasis. We evaluated the metabolic stone profile in patients presenting for parathyroid surgery and determined whether there were differences between those with and without a history of urinary stones. MATERIALS AND METHODS: A total of 60 patients with primary hyperparathyroidism presenting for parathyroid surgery were prospectively enrolled in the study, of whom 54 completed a preoperative 24-hour urine collection. Of these 54 patients 27 completed an additional postoperative 24-hour urine collection. We compared preoperative metabolic urinary profiles and serum calcium in patients with primary hyperparathyroidism with and without a history of urolithiasis. In addition, we compared changes in urine metabolic profiles that occurred after successful parathyroid surgery. RESULTS: There were no significant differences in preoperative 24-hour urine metabolic profiles or serum calcium between patients who had primary hyperparathyroidism with and without a history of urolithiasis. Compared to preoperative levels after successful parathyroid surgery there were significant decreases in serum calcium (10.8 to 9.3 mg/dl, p <0.001), urinary calcium (319 to 156 mg per day, p <0.01) calcium oxalate supersaturation (8.6 to 5.7, p = 0.016) and calcium phosphate supersaturation (1.6 to 0.9, p = 0.002) in the 27 patients who completed a postoperative 24-hour urine collection. CONCLUSIONS: Other etiological factors must exist that cause some patients with primary hyperparathyroidism to form stones, while most never have stones despite profound hypercalcemia and hypercalciuria. Routine 24-hour urine evaluation cannot predict which patients with primary hyperparathyroidism will have kidney stones. PMID- 19296982 TI - Late diagnosis of primary hyperoxaluria type 2 in the adult: effect of a novel mutation in GRHPR gene on enzymatic activity and molecular modeling. AB - PURPOSE: Genetic causes of nephrolithiasis are underestimated. Primary hyperoxaluria type 2 is a rare autosomal recessive disease caused by mutations in the GRHPR gene, leading to an accumulation of oxalate and L-glycerate with recurrent kidney stone formation and nephrocalcinosis, and the later development of renal failure and systemic oxalate depositions. We studied the effects of a novel GRHPR mutation on GRHPR enzymatic activity and molecular modeling. MATERIALS AND METHODS: Genomic DNA from a 50-year-old male with a late diagnosis of primary hyperoxaluria type 2 was extracted, analyzed and compared with the established human GRHPR gene sequence. Restriction enzyme analysis of the patient, 30 healthy controls and 30 patients with nephrolithiasis of various causes was done to confirm the presence of the mutation. GRHPR activity was analyzed by site directed mutagenesis of WT and mutant clones. We studied the effects of the mutation on enzymatic molecular modeling. RESULTS: We found the novel homozygous single missense mutation A975G in exon 9, creating an amino acid change from asparagine to aspartic acid in position 312. No mutations were detected in restriction enzyme analysis in all 30 healthy controls and 30 patients with nephrolithiasis of various causes. Transfected cells with the mutant clone showed abolished GRHPR activity. Molecular modeling studies revealed that the mutation was likely to disrupt the correct folding of the GRHPR substrate binding domain, hence affecting the enzyme active site. CONCLUSIONS: Primary hyperoxaluria type 2 should be considered in patients at adult stone clinics who have had a history of nephrolithiasis since childhood, especially in those with consanguineous parents. Biochemical analysis followed by mutation identification should be the approach for making the definitive diagnosis of primary hyperoxaluria type 2. PMID- 19296983 TI - Cost-effectiveness of botulinum toxin a versus anticholinergic medications for idiopathic urge incontinence. AB - PURPOSE: We assessed the cost-effectiveness of botulinum toxin A injection compared to anticholinergic medications for the treatment of idiopathic urge incontinence. MATERIALS AND METHODS: A Markov decision analysis model was developed to compare the costs in 2008 U. S. dollars and effectiveness in quality adjusted life-years of botulinum toxin A injection and anticholinergic medications. The analysis was conducted from a societal perspective with a 2-year time frame using 3-month cycles. The primary outcome was the incremental cost effectiveness ratio, defined as the difference in cost (botulinum toxin A cost- anticholinergic cost) divided by the difference in effectiveness (botulinum toxin A quality adjusted life-years--anticholinergic quality adjusted life-years). RESULTS: While the botulinum strategy was more expensive ($4,392 vs $2,563) it was also more effective (1.63 vs 1.50 quality adjusted life-years) compared to the anticholinergic regimen. The calculated incremental cost-effectiveness ratio was $14,377 per quality adjusted life-year, meaning that botulinum toxin A cost $14,377 per quality adjusted life-year gained. A strategy is often considered cost-effective when the incremental cost-effectiveness ratio is less than $50,000 per quality adjusted life-year. Given this definition botulinum toxin A is cost effective compared to anticholinergics. To determine if there are situations in which anticholinergics would become cost-effective we performed sensitivity analyses. Anticholinergics become cost-effective if compliance exceeds 75% (33% in the base case) and if the botulinum toxin A procedure cost exceeds $3,875 ($1,690 in the base case). For the remainder of the sensitivity analyses botulinum toxin A remained cost-effective. CONCLUSIONS: Botulinum toxin A injection was cost-effective compared to anticholinergic medications for the treatment of refractory urge incontinence. Anticholinergics become cost-effective if patients are highly compliant with medications or if the botulinum procedure costs increase substantially. PMID- 19296985 TI - Synchronous urethral stricture reconstruction via 1-stage ascending approach: rationale and results. AB - PURPOSE: We present our experience with the reconstruction of synchronous urethral strictures. MATERIALS AND METHODS: Of 482 anterior urethroplasties performed by a single surgeon between 1997 and 2008 we identified and reviewed 30 patients who underwent reconstruction for multiple separate strictures. An ascending approach from distal to proximal was used and all repairs were completed at 1 stage. A total of 13 combinations of techniques were used to complete the repairs. A 2-phase technique was used in which the patient remained supine during buccal mucosa harvest and repair of strictures distal to the penoscrotal junction, and was then repositioned into the high lithotomy position as needed for stricture repair in the bulbar urethra. In each case normal intervening urethra was preserved intact. The number, length and location of strictures, operative time and patient outcomes were evaluated. RESULTS: No position related complications occurred during or after surgery despite a mean operative time of 4.5 hours (range 2.5 to 6.4). No infectious wound complications were reported despite repositioning the legs to the high lithotomy position. Three patients (10%) were known to have required treatment for recurrent stricture after surgery. CONCLUSIONS: One-stage reconstruction for synchronous urethral strictures may be safely and effectively performed using a systematic, ascending reconstructive approach with creative application of tissue transfer techniques. Decreasing patient time in the high lithotomy position appears to prevent related lower extremity complications. PMID- 19296986 TI - Evaluating use patterns of and adherence to medications for benign prostatic hyperplasia. AB - PURPOSE: We investigated adherence to benign prostatic hyperplasia medications in a California Medicaid population. MATERIALS AND METHODS: Using California Medicaid data on 1995 to 2004 we identified adult males 40 years old or older with 1 or more diagnosis and 2 or more prescription fills for benign prostatic hyperplasia. Patients with 2 fills on the same day were assigned to the multiple medication cohort. Adherence was measured using the medication possession ratio for the index medication and the proportion of days covered for any benign prostatic hyperplasia medication. Patients with a medication possession ratio or proportion of days covered of 0.8 or greater were considered adherent. A Cox proportional hazards model was used to assess the relative hazards associated with discontinuation. Multiple logistic regression was used to investigate factors associated with nonadherence or a benign prostatic hyperplasia related procedure. RESULTS: Of the total population of 2,640 men 40% were adherent with any benign prostatic hyperplasia medication. A significantly greater proportion of patients using multiple medications and finasteride were adherent with any benign prostatic hyperplasia medication (62% and 55%, respectively, p <0.0001). Doxazosin, terazosin and tamsulosin use was associated with nonadherence (p = 0.008, 0.04 and 0.03, respectively). Younger patients and those changing medications were more likely to discontinue (p = 0.01 and <0.0001), while patients using multiple medications and those experiencing a gap were at lower risk for discontinuation (p = 0.01 and <0.0001, respectively). Predictors of a procedure included an index prescription in 1999 or later, a urologist visit and nonadherence to any benign prostatic hyperplasia medication (p = 0.01, <0.0001 and <0.0001, respectively). CONCLUSIONS: Adherence to alpha-blockers was less than adherence to finasteride or multiple medications and nonadherence was significantly associated with a procedure. Interventions focused on improving adherence to benign prostatic hyperplasia medications are clearly needed. PMID- 19296988 TI - The significance of 99mtechnetium dimercapto-succinic acid renal scan in children with spina bifida during long-term followup. AB - PURPOSE: (99m)Technetium dimercapto-succinic acid renal scans are ideal for demonstrating renal scarring in children with spina bifida. However, doubt persists about the need for routine application. We assessed the associations among abnormal (99m)technetium dimercapto-succinic acid renal scans, vesicoureteral reflux and urodynamic findings in patients with spina bifida during long-term followup. MATERIALS AND METHODS: We retrospectively reviewed the records of 64 patients with spina bifida followed at our center. All patients were older than 10 years (mean 15.8, range 10 to 23). Dimercapto-succinic acid renal scans were considered abnormal with differential function of less than 40% or focal defects. Patient age, gender, previous febrile urinary tract infections, positive vesicoureteral reflux history, timing of clean intermittent catheterization initiation and the latest urodynamic findings were noted. Patients were grouped based on normal/abnormal scan results. Statistical analysis included univariate and multivariate regression analyses and chi-square tests. RESULTS: A total of 16 patients (25%) had abnormal scans. Mean patient age, male to-female ratio, leak point pressure, bladder compliance and timing of clean intermittent catheterization initiation did not differ between groups. Rates of previous febrile urinary tract infections differed significantly (11 of 16 in the abnormal group vs 9 of 48 in the normal group, p <0.01), as did positive reflux history (100% vs 31%, p <0.01). No patient with a negative reflux history had an abnormal dimercapto-succinic acid renal scan. Multivariate analysis identified previous febrile urinary tract infections as a significant risk factor for an abnormal scan. CONCLUSIONS: A positive vesicoureteral reflux history and febrile urinary tract infections were associated with abnormal dimercapto-succinic acid renal scan in followup of patients older than 10 years with spina bifida. Thus, these factors are indicators of proactive evaluation of renal function using dimercapto-succinic acid renal scanning. PMID- 19296991 TI - The relationship of Irgarol and its major metabolite to resident phytoplankton communities in a Maryland marina, river and reference area. AB - The objectives of this study were to: (1) measure water column concentrations of Irgarol 1051 and its major metabolite GS26575 annually (2004-2006) during mid June and mid-August at 14 sites in a study area comprised of three sub-regions chosen to reflect a gradient in Irgarol exposure (Port Annapolis marina, Severn River and Severn River reference area); (2) use a probabilistic approach to determine ecological risk of Irgarol and its major metabolite in the study area by comparing the distribution of exposure data with toxicity-effects endpoints; and (3) measure both functional and structural resident phytoplankton parameters concurrently with Irgarol and metabolite concentrations to assess relationships and determine ecological risk at six selected sites in the three study areas described above. The three-year summer mean Irgarol concentrations by site clearly showed a gradient in concentrations with greater values in Back Creek (400-500ng/L range), lower values in the Severn River sites near the confluence with Back Creek (generally values less than 100ng/L) and still lower values (<10ng/L) at the Severn River reference sites at the confluence with Chesapeake Bay. A similar spatial trend, but with much lower concentrations, was also reported for GS26575. The probability of exceeding the Irgarol plant 10th centile of 193ng/L and the microcosm NOEC (323ng/L) suggested high ecological risk from Irgarol exposure at Port Annapolis marina sites but much lower risk at the other sites. There were no statistically significant differences among the three site types (marina, river and reference) with all years combined or among years within a site type for the following functional and structural phytoplankton endpoints: algal biomass, gross photosynthesis, biomass normalized photosynthesis, chlorophyll a, chlorophyll a normalized photosynthesis and taxa richness. Therefore, based on the above results, Irgarol adverse effects predicted from the plant 10th centile and the microcosm NOEC in the high Irgarol exposure area (Back Creek/Port Annapolis marina) were not confirmed with the actual field data for the receptor species (phytoplankton). These results also highlight the importance of unconfined field studies with a chemical gradient in providing valuable information regarding the responses of resident phytoplankton to herbicides. PMID- 19296992 TI - First record of Prorocentrum lima (Dinophyceae) inside harbor areas and along the Abruzzo region coast, W Adriatic. AB - Prorocentrum lima (Ehrenberg) Dodge has been found for the first time during the summer of 2007 inside Ortona harbor and along the coast of the Abruzzo region, a slightly eutrophic area influenced by runoff from a nearby river. The investigations were conducted in two harbors and at six coastal sampling stations. Samplings were conducted using a phytoplankton net and with a pump. Average P. lima cellular concentrations were 3.2 x 10(4)cells L(-1). Other well known toxic and potentially toxic phytoplankton species have been considered. The number of toxic cells from net samples were higher than the numbers of toxic cell samples collected without the net. Occurrences of P. lima with abiotic factors revealed that temperature was positively correlated with P. lima abundance (p=0.01), while salinity was highly negatively correlated with P. lima presence (p=0.001). The total phytoplankton community was studied. PMID- 19296995 TI - [Focal subarachnoid hemorrhage and cerebral amyloid angiopathy: a non-fortuitous association]. AB - INTRODUCTION: Cerebral amyloid angiopathy is a degenerative angiopathy due to amyloid deposits in the walls of the meningeal and cortical vessels. It is considered as a major cause of cerebral hemorrhage to the elderly. It was recently demonstrated that the association of focal meningeal bleedings and cerebral hemorrhage is very suggestive of cerebral amyloid angiopathy. In contrast, the links between subarachnoid hemorrhage and amyloid angiopathy are less well-known. CASES REPORTS: We report nine cases of cerebral amyloid angiopathy. The clinical presentation was variable, but all had at least one inaugural meningeal bleeding. As cortico-meningeal biopsies were not performed the Boston criteria were used to establish the diagnosis. CONCLUSION: Cerebral amyloid angiopathy is an underestimated cause of subarachnoid hemorrhage. Our observations show that this diagnosis should be evoked when focal meningeal bleeding occurs without head trauma or when focal subarachnoid hemorrhage is followed by a subcortical hematoma in an elderly subject. PMID- 19296996 TI - [Expert workshop on status epilepticus: an example of fruitful collaboration]. PMID- 19296997 TI - [The French version of the FOUR score: A new coma score]. AB - BACKGROUND: Comatose state is a major cause for admission to the intensive care unit. The most commonly used assessment score is the Glasgow coma scale (GCS). Although widely accepted, this score has several limitations. Recently, the full outline of unresponsiveness score (FOUR) has been validated and tested as reliable as the GCS. METHODS: We translated this score in French and tested its reliability in a neurological critical care unit. This study included eight critical care patients and eight intensive care patients. The patients were successively evaluated by two neurologists, four experienced nurses and five inexperienced nurses; a total of 176 evaluations were performed. The weighted kappa (kappa(W)) was used to determine the reliability of the evaluation for both the FOUR score and the GCS. RESULTS: The mean age of the patients was 62 years. The interobserver reliability of the French version of the FOUR score was high (kappa(W)=0.86; IC 95%: 0.83-0.89) comparable to that of the GCS (kappa(W)=0.85; IC 95%: 0.82-0.88). CONCLUSION: The French version of the FOUR score has an excellent interobserver reliability. This score is easy to perform and well accepted, only requiring simple and short training. PMID- 19296999 TI - [Emergency and intensive care unit management of status epilepticus Societe de reanimation de langue francaise Experts Recommendations: the transient triumph of the followers of Sisyphus?]. PMID- 19296998 TI - [Emergency and intensive care unit management of status epilepticus in adult patients and children (new-born excluded). Societe de reanimation de langue francaise experts recommendations]. PMID- 19297000 TI - Hydrogenosomes under microscopy. AB - A hydrogenosome is a hydrogen-producing organelle, evolutionary related to mitochondria and is found in Parabasalia protozoa, certain chytrid fungi and certain ciliates. It displays similarities to and differences from mitochondria. Hydrogenosomes are spherical or slightly elongated organelles, although very elongated hydrogenosomes are also found. They measure from 200 nm to 1 microm, but under stress conditions can reach up to 2 microm. Hydrogenosomes are surrounded by two closely apposed membranes and present a granular matrix. Cardiolipin has been detected in their membranes, and frataxin, which is a conserved mitochondrial protein involved in iron metabolism, was also recently found. Hydrogenosomes have one or multiple peripheral vesicles, which incorporate calcium. The peripheral vesicle can be isolated from the hydrogenosomal matrix and can be considered as a distinct hydrogenosomal compartment. Dysfunctional hydrogenosomes can be removed by an autophagic process and further digested by lysosomes. Hydrogenosomes divide in three different ways, like mitochondria, by segmentation, partition and the heart form. They may divide at any phase of the cell cycle. Nucleoid or electron dense deposits found in hydrogenosomes can be considered artifacts or dysfunctional hydrogenosomes. The hydrogenosome does not contain a genome, although DNA has already been detected in one anaerobic ciliate. Hydrogenosomes can be considered as good drug targets since their metabolism is distinct from mitochondria. PMID- 19297001 TI - Propagation of thickness-twist waves in a piezoelectric ceramic plate with unattached electrodes. AB - We analyze the propagation of thickness-twist waves in an unbounded piezoelectric ceramic plate with air gaps between the plate surfaces and two electrodes. These waves are also called anti-plane or shear-horizontal waves with one displacement component only. An exact solution is obtained from the equations of the linear theory of piezoelectricity. Dispersion relations of the waves are obtained and plotted. Results show that the wave frequency or speed is sensitive to the air gap thickness. This effect can be used to manipulate the behavior of the waves and has implications in acoustic wave devices. PMID- 19297002 TI - Long-term ammonia removal in a coconut fiber-packed biofilter: analysis of N fractionation and reactor performance under steady-state and transient conditions. AB - A comprehensive study of long-term ammonia removal in a biofilter packed with coconut fiber is presented under both steady-state and transient conditions. Low and high ammonia loads were applied to the reactor by varying the inlet ammonia concentration from 90 to 260 ppm(v) and gas contact times ranging from 20 to 36 s. Gas samples and leachate measurements were periodically analyzed and used for characterizing biofilter performance in terms of removal efficiency (RE) and elimination capacity (EC). Also, N fractions in the leachate were quantified to both identify the experimental rates of nitritation and nitratation and to determine the N leachate distribution. Results showed stratification in the biofilter activity and, thus, most of the NH(3) removal was performed in the lower part of the reactor. An average EC of 0.5 kg N-NH(3)m(-3)d(-1) was obtained for the whole reactor with a maximum local average EC of 1.7 kg N-NH(3)m(-3)d( 1). Leachate analyses showed that a ratio of 1:1 of ammonium and nitrate ions in the leachate was obtained throughout steady-state operation at low ammonia loads with similar values for nitritation and nitratation rates. Low nitratation rates during high ammonia load periods occurred because large amounts of ammonium and nitrite accumulated in the packed bed, thus causing inhibition episodes on nitrite-oxidizing bacteria due to free ammonia accumulation. Mass balances showed that 50% of the ammonia fed to the reactor was oxidized to either nitrite or nitrate and the rest was recovered as ammonium indicating that sorption processes play a fundamental role in the treatment of ammonia by biofiltration. PMID- 19297003 TI - Investigation and optimization of a passively operated compost-based system for remediation of acidic, highly iron- and sulfate-rich industrial waste water. AB - A passively operated multi-stage bioremediation system utilizing composted agricultural waste products and an artificial wetland system was found to be effective for purification of acidic, iron- and sulfate-rich waste water derived from titanium mineral processing. The main microbial players involved in the remediation system processes and the dynamics were investigated; mineral processing waste water-filled sludge dams possessed stable microbial communities that included Acidithiobacillus, Desulfurella, and acidophilic, anaerobic fermenters of the order Bacteroidales. These groups were enriched in a subsequent potato waste-based iron mobilization pre-treatment stage. Within downstream reduction treatment stages ("reduction cells"), compost/straw decomposition and associated sulfur/sulfate and iron reduction were carried out by a complex mix of aerobic and anaerobic bacteria. The efficaciousness of the system without replacement of the compost was found to steadily decline following 2 years of operation and corresponded with the reduction cell communities becoming simultaneously more diverse and homogenous. Microcosm-based experiments demonstrated that operational declines were due to unsustained supply of suitable labile carbon sources combined with spatial heterogeneity within the layered design of the reduction stage of the system resulting in inadequate redox conditions. Temperature was not found to be a critical performance factor in the range of 10-25 degrees C. Application of a combined emulsified oil/molasses amendment was found to be highly effective in promoting a microbial community capable of remediating waste water with high iron and sulfate levels. Acidophilic members of the order Bacteroidales were found to be critical in the investigated remediation system, providing organic donors for subsequent metal and sulfur transformations and could have a broader ecological significance than previously suspected. PMID- 19297004 TI - Modeling anaerobic digestion of aquatic plants by rumen cultures: cattail as an example. AB - Despite of the significance of the anaerobic digestion of lignocellulosic materials, only a limited number of studies have been carried out to evaluate the lignocellulosic digestion kinetics, and information about the modeling of this process is limited. In this work, a mathematical model, based on the Anaerobic Digestion Model No.1 (ADM1), was developed to describe the anaerobic conversion of lignocellulose-rich aquatic plants, with cattail as an example, by rumen microbes. Cattail was fractionated into slowly hydrolysable fraction (SHF), readily hydrolysable fraction (RHF) and inert fraction in the model. The SHF was hydrolyzed by rumen microbes and resulted in the production of RHF. The SHF and RHF had different hydrolysis rates but both with surface-limiting kinetics. The rumen microbial population diversity, including the cattail-, butyrate-, acetate- and H(2)-degraders, was all incorporated in the model structure. Experiments were carried out to identify the parameters and to calibrate and validate this model. The simulation results match the experimental data, implying that the fractionation of cattail into two biodegradation parts, i.e., SHF and RHF, and modeling their hydrolysis rate with a surface-limiting kinetics were appropriate. The model was capable of simulating the anaerobic biodegradation of cattail by the rumen cultures. PMID- 19297005 TI - High prevalence of chronic myocarditis in dilated cardiomyopathy referred for left ventriculoplasty: expression of tenascin C as a possible marker for inflammation. AB - The objectives of this study were to analyze the incidence of chronic myocarditis in dilated cardiomyopathy and to evaluate the diagnostic value of tenascin C for assessing inflammatory activity in the resected myocardium. Dilated cardiomyopathy patients with chronic myocarditis have a poor clinical outcome despite recent advances in medical treatments. Therefore, a precise diagnosis of inflammatory activity is critical to ensuring appropriate therapy. Tenascin C is an extracellular matrix glycoprotein that plays an important role in tissue remodeling in various heart diseases. Myocardial samples obtained during left ventriculoplasty from 64 patients (50 +/- 13 years, 56 men and 8 women) with dilated cardiomyopathy were examined by immunostaining for tenascin C. Histologic diagnosis was based on the Dallas criteria modified by the International Society and Federation of Cardiology task force. Nine cases (14%) had active myocarditis, 21 (33%) had borderline myocarditis, and 34 (53%) had no myocarditis. Intense tenascin C expression was observed at the site of active inflammation, with abundant cell accumulation, and in organized granulation tissue during the resolving phase but not in scar tissue during the healing phase. The ratio of tenascin C-positive area to the whole myocardium in the active and borderline myocarditis groups was significantly greater than that in the noninflammatory group. These findings suggest a high prevalence of chronic myocarditis in dilated cardiomyopathy patients and that tenascin C may prove to be a useful marker for distinguishing inflammatory cardiomyopathy from other types of dilated cardiomyopathy. PMID- 19297006 TI - A paucity of colonic enteroendocrine and/or enterochromaffin cells characterizes a subset of patients with chronic unexplained diarrhea/malabsorption. AB - A generalized absence of enteroendocrine cells characterizes 2 diarrheal/malabsorptive diseases, namely, enteroendocrine cell dysgenesis and autoimmune polyglandular syndrome 1. However, it is not routine for pathologists to examine mucosal biopsies for enteroendocrine cells in cases of chronic diarrheal illness. Our primary aim was to prospectively examine colonic mucosa for loss of enteroendocrine cells using chromogranin A immunohistochemistry for diagnostic purposes. Our secondary aim was to investigate enterochromaffin cells as a subset of enteroendocrine cells, using serotonin (5HT) immunohistochemistry; we hypothesized that other causes of diarrhea due to loss of enteroendocrine cell subsets are missed by evaluating enteroendocrine cells alone. Our approach was limited to patients with chronic unexplained diarrhea partly selected by referring physicians who considered the patients problematic. Seven problematic patients with reduced enteroendocrine or enterochromaffin cells were collected over a 9-month period and placed in group A. Three group A patients demonstrated reduced enteroendocrine cells relative to controls, and they were later diagnosed as having enteroendocrine cell dysgenesis (n = 1) and autoimmune polyglandular syndrome 1 (n = 2). Four group A patients had reduced enterochromaffin cells but normal enteroendocrine cells. These 4 patients had conditions such as congenital diarrhea, mild graft-versus-host disease, acquired childhood chronic diarrhea, and diarrhea post lung transplant. The reduced enterochromaffin cells in the graft-versus-host disease patient inspired a third aim, that is, to investigate whether a loss of enterochromaffin cells would be a generalized defect seen in patients with mild colonic graft-versus-host disease (group B). However, no loss of enterochromaffin cells was detected in group B. Two methods of enumerating endocrine cells were used and demonstrated 67% agreement. PMID- 19297007 TI - Pagetoid spread to oral mucosa from submandibular gland salivary duct carcinoma: a case report. AB - Salivary duct carcinoma is a high-grade malignant tumor arising predominantly in the major salivary glands. We describe herein a rare presentation of salivary duct carcinoma arising in the submandibular gland and showing pagetoid spread to the oral mucosa. A 46-year-old man was admitted with a 2-year history of swelling in the floor of the mouth. Resection of the submandibular gland tumor was followed by mucosal biopsy around the aperture of the right Wharton's duct. The tumor recurred in the oral mucosa with pagetoid spread and extended widely in the oral cavity for the following 10 years, despite chemo- and radiotherapy. Mucosal spread, although a relatively remote possibility, can occur in the setting of salivary duct carcinoma. PMID- 19297008 TI - Plasmacytoid urothelial carcinoma of the bladder. AB - In this report, we present the clinicopathologic features of 11 cases of the plasmacytoid variant of urothelial carcinoma. This is a rare variant of bladder cancer recognized by the current World Health Organization classification of urologic tumors. The plasmacytoid component varied from 30% to 100% of the tumor specimen; in 8 cases, the plasmacytoid component comprised greater than 50% of the tumor with 2 cases showing pure plasmacytoid carcinoma. The architectural pattern of the tumor varied from solid expansile nests with noncohesive cells to mixed solid and alveolar growth; a streaking discohesive architecture was additionally present in 2 cases (18%). At histology, the individual tumor cells had an eccentrically placed nucleus and abundant eosinophilic cytoplasm reminiscent of plasma cells. Most neoplastic cells had nuclei of low to intermediate nuclear grade with occasional nuclear pleomorphism. Seven of 9 mixed cases had concurrent conventional high-grade urothelial carcinoma, and the remaining 2 cases presented features of nested or micropapillary urothelial carcinoma. Small intracytoplasmic vacuoles were variably present in all cases. All patients had advanced stage cancer (>pT3), and 8 (73%) had lymph node metastasis. Immunohistochemical staining demonstrated that both plasmacytoid and associated conventional urothelial carcinoma were positive for cytokeratins 7, 20, and AE1/AE3 and epithelial membrane antigen; CD138 was positive in 3 cases. Follow-up information was available in all cases (range, 2-16 months; mean, 7 months). Nine of the patients died of disease from 2 to 11 months, and 2 patients were alive with disease at 8 and 16 months. In summary, plasmacytoid variant of urothelial carcinoma is an aggressive variant associated with poor prognosis that presents at an advanced clinical stage. In limited samples, it may be misdiagnosed as chronic cystitis or plasmacytoma, a pitfall further compounded by CD138 expression in some cases. Morphological distinction from other malignant neoplasms with plasmacytoid phenotype is critical for its clinical management. PMID- 19297009 TI - Optimal running speed and the evolution of hominin hunting strategies. AB - Recent discussion of the selective pressures leading to the evolution of modern human postcranial morphology, seen as early as Homo erectus, has focused on the relative importance of walking versus running. Specifically, these conversations have centered on which gait may have been used by early Homo to acquire prey. An element of the debate is the widespread belief that quadrupeds are constrained to run at optimally efficient speeds within each gait, whereas humans are equally efficient at all running speeds. The belief in the lack of optimal running speeds in humans is based, however, on a number of early studies with experimental designs inadequate for the purpose of evaluating optimality. Here we measured the energetic cost of human running (n=9) at six different speeds for five minutes at each speed, with careful replicates and controls. We then compared the fit of linear versus curvilinear models to the data within each subject. We found that individual humans do, in fact, have speeds at which running is significantly less costly than at other speeds (i.e., an optimal running speed). In addition, we demonstrate that the use of persistence hunting methods to gain access to prey at any running speed, even the optimum, would be extremely costly energetically, more so than a persistence hunt at optimal walking speed. We argue that neither extinct nor extant hominin populations are as flexible in the chosen speeds of persistence hunting pursuits as other researchers have suggested. Variations in the efficiency of human locomotion appear to be similar to those of terrestrial quadrupeds. PMID- 19297010 TI - Evaluation on the toxicity of nanoAg to bovine serum albumin. AB - Measuring protein damage by nanomaterials may give insight into the mechanisms of toxicity of nanomaterials. The toxic effects of nanoAg on bovine serum albumin (BSA) were thoroughly studied using fluorescence spectroscopy, ultraviolet visible absorption spectroscopy, resonance light scattering spectroscopy (RLS), circular dichroism spectroscopy (CD) and transmission electron microscopy (TEM). NanoAg had obvious toxic effects on BSA: nanoAg could increase the amount of helix and decrease the beta sheet structure, leading to a loosening of the protein skeleton. In the loose structure, internal hydrophobic amino acids are exposed and the characteristic fluorescence of BSA is obviously quenched. When the ratio of nanoAg and BSA increased to 1: 96 (wt/wt), the impact of nanoAg on the spectral properties leveled off. The RLS spectrum, TEM, CD spectra and electrophoresis results showed that BSA had destroyed the double-layer structure of nanoAg and covered its surface, generating a BSA-nanoAg complex held together by van der Waals and electrostatic forces. This paper provides a new perspective and method for determining the toxic effects of nanoAg on biological macromolecules. PMID- 19297011 TI - The effect of high circulating estradiol levels on thrombin generation during in vitro fertilization. PMID- 19297012 TI - Extragastrointestinal stromal tumor arising in the rectovaginal septum: report of an unusual case with literature review. AB - BACKGROUND: Extragastrointestinal stromal tumor (EGIST) is a rare tumor that shows features of gastrointestinal stromal tumor (GIST) outside the gastrointestinal tract, but EGIST arising in the rectovaginal septum is extremely rare. CASE: We report a unique case of a 42-year-old woman with EGIST in rectovaginal septum. The tumor was excised locally. The patient is healthy without evidence of recurrence 11 months after surgery. CONCLUSION: The clinical features and treatment of EGIST are not well known because few cases have been reported. Pathologically, immunohistochemistry with antibody against c-kit protein (CD117) is valuable for diagnosis of EGIST. Mitotic rate is the most reliable prognostic factor, and mutations in KIT may be a clinically useful adjunct marker in the evaluation. Surgery is the most effective treatment, and long-term follow-up is necessary. PMID- 19297013 TI - A comparison of quality of life between vulvar cancer patients after sentinel lymph node procedure only and inguinofemoral lymphadenectomy. AB - OBJECTIVES: The SLN-procedure has been introduced in vulvar cancer treatment to reduce morbidity and thereby improve quality of life. Aim of this study was to compare quality of life in vulvar cancer patients who were treated with a SLN procedure only to those who underwent inguinofemoral lymphadenectomy. Moreover, it was evaluated what patients would advise relatives on the application of the SLN-procedure in light of possible false negative results. METHODS: Patients who participated in the GROningen INternational Study on Sentinel nodes in Vulvar cancer (GROINSS-V) were invited to fill out three questionnaires: the EORTC QLQ C30, a vulvar specific questionnaire and a questionnaire about the opinion of patients on new treatment options. Patients who only underwent SLN-procedure were compared to those who subsequently underwent inguinofemoral lymphadenectomy because of a positive SLN. RESULTS: With a response rate of 85%, 35 patients after the SLN-procedure and 27 patients after inguinofemoral lymphadenectomy filled out the questionnaires. No difference in overall quality of life was observed between the two groups. The major difference was the increase in complaints of lymphedema of the legs after inguinofemoral lymphadenectomy. The majority of patients would advise the SLN-procedure to relatives. Patients after inguinofemoral lymphadenectomy were more reserved concerning the acceptable false negative rate of a new diagnostic procedure. CONCLUSIONS: Patients who underwent the SLN-procedure report less treatment related morbidity compared to those who underwent inguinofemoral lymphadenectomy. However, this did not influence overall quality of life. Furthermore, patients who underwent inguinofemoral lymphadenectomy are more reserved in advising the SLN-procedure to relatives. PMID- 19297014 TI - Routine follow-up intervals in patients with high-grade squamous intraepithelial lesions (HSIL) and free excision margins can safely be increased in the first two years after Large Loop Excision of the Transformation Zone (LLETZ). AB - OBJECTIVE: To assess the compliance of HSIL patients to the national Dutch routine follow-up protocol in the first 2 years after LLETZ and to determine if based on the status of excision margins, follow-up intervals could be modified. METHODS: A prospective cohort study was performed in patients, referred because of an abnormal Pap smear between 1996 and 2004 and treated for HSIL with LLETZ. The Dutch national routine follow-up protocol orders a Pap smear after 6, 12 and 24 months, respectively. Follow-up results were completed by using PALGA, the nationwide network and registry of histo- and cytopathology in the Netherlands. To assess compliance to the follow-up protocol, adequate follow-up was defined as three cervical smears taken after 6 (+/-3), 12 (+/-3) and 24 (+/-3) months, respectively. RESULTS: Compliance to the first 2 years follow-up protocol declined from 86.2% to 64.8% to 51.2% for first, second and third follow-up cervical smears, respectively. Patients with involved excision margins had a three times higher overall risk of developing a subsequent HSIL after LLETZ as compared to patients with free excision margins (HR: 3.2, 95% CI=1.3-7.9, p=0.01). Risk for diagnosing HSIL during the first 12 months of follow-up for patients with free excision margins was only 1%. CONCLUSIONS: Compliance to the Dutch national routine follow-up protocol in HSIL patients after LLETZ is only moderate. For HSIL patients with free excision margins after LLETZ the first cytological follow-up interval can safely be increased to 12 months. PMID- 19297015 TI - Sonomorphology of the reproductive tract in male and pregnant and non-pregnant female Rothschild's giraffes (Giraffa camelopardalis rotschildi). AB - The application of real-time-B-mode ultrasonography to wild and zoo animal medicine has been shown to improve the understanding of reproductive physiology in many species. Ultrasound technology is especially helpful for monitoring urogenital health, which in turn has advantages for giraffe breeding and welfare in captivity. This study aimed to ultrasonographically describe the genital organs of reproductively healthy male and female giraffes. Through the use of a restrainer, repeated rectal ultrasound examinations were performed over a 2 year period in 2.6 Rothschild's giraffes. Changes in ovarian activity were monitored throughout four different reproductive stages in the females and included immature, mature-cycling, pregnancy, post-partum-period. In the immature giraffes the ovaries showed multiple follicles of which larger ones luteinized to form pseudo-corpora lutea. By comparison, in the mature giraffes the dominant follicle reached an ovulatory diameter of 18.5+/-0.89 mm. After ovulation, a single corpus luteum rapidly formed and reached a maximum diameter of 33.0+/-2.4mm on average. Pregnancy was detected for the first time by the embryonic vesicle, visualized around 28 days post copulation. Follicular development remained ongoing during early pregnancy. In the males, as in other ruminants, the bulbourethral glands and the seminal vesicles were prominent, whereas the prostate gland was indistinct. Knowledge about the reproductive tract morphology and physiology is necessary for diagnosing medical disorders and abnormalities in giraffes. The aim of this study was to help consolidate the current knowledge on basic reproductive parameters for this species. PMID- 19297016 TI - A study of the adhesion between dental cement and dentin using a nondestructive acoustic microscopy approach. AB - OBJECTIVES: The goal of the present study was to investigate the potential for acoustic microscopy techniques to characterize the cement-dentin interface in restored teeth. METHODS: Special flat-parallel specimens and whole extracted teeth with restorations were scanned using a high-frequency (50 MHz) focused ultrasonic transducer. Visual acoustic images (B- and C-scans) of the cement dentin interface were obtained nondestructively, analyzed and compared with optical images taken after the samples were cut along the scanning axis. The shear bonding strength of a subsection of specimens was tested in a Lloyd material testing machine. RESULTS: An essential distinction between the acoustical properties associated with good and failed bonding has been shown. In the case of failed adhesion, the ultrasound signal reflection from the cement dentin interface is up to four to seven times higher in magnitude than in the case of good bonding. The comparison of the ultrasound imaging data with the data obtained using an optical microscope revealed a strong correspondence with the acoustical and optical results with respect to the presence, position and dimensions of the defects. The specimens showing higher ultrasound reflection from cement/dentin interface have also shown lower shear bonding strength. SIGNIFICANCE: The results demonstrate that acoustic scanning with a high frequency focused ultrasonic probe is a valuable method for nondestructive morpho mechanical analysis of cement/dentin interface for either experimental models or whole restored teeth. An appropriately expanded approach can be widely used for the pre-clinical evaluation of dental materials. Further, this method may prove beneficial in the design of new diagnostic ultrasound devices and techniques for use within clinical dentistry. PMID- 19297017 TI - The basic and clinical science of twin-twin transfusion syndrome. AB - Twin-twin transfusion syndrome (TTTS) is a fascinating condition in which fetuses of identical genotype adopt discordant cardiovascular phenotypes, secondary to unbalanced placental inter-twin transfusion. Flow along the primary units of inter-twin transfusion, unidirectional arteriovenous anastomoses, can be as high as litres/day each, and TTTS develops when the placenta has insufficient compensatory counter-transfusional anastomoses. The initial phenotype reflects dysvolaemia, with added contributions from uteroplacental insufficiency in the donor, and raised afterload with diastolic dysfunction secondary to discordant endothelin and placentally derived renin-angiotensin system effectors in the recipient. Endoscopic laser ablation of placental anastomoses has become the primary treatment modality, supported by a randomised trial showing improved survival and short but not long-term neurological morbidity. Its uptake has facilitated comparative pre- and post-laser studies, which provide considerable insight into the pathophysiology. Despite the therapeutic advance, placental laser remains associated with a 25% incidence of fetal death within a week, and a 10% risk each of recurrence and twin anaemia/polycythaemia sequence due to residual anastomoses. In Stage I, high rates of non-progression with more conservative management have resulted in therapeutic equipoise as to whether laser is indicated primarily or only for progressive disease. The challenge ahead lies in improving double intact survival rates, which in addition to randomised trials will require technical advances, better understanding of the circulatory pathophysiology and more sophisticated surveillance tools. PMID- 19297018 TI - Ionotropic purinergic receptor P2X4 is involved in the regulation of chondrogenesis in chicken micromass cell cultures. AB - We have previously demonstrated that elevation of free cytosolic Ca(2+) concentration at the time of differentiation of chondroblasts was mainly due to a Ca(2+) influx and it was indispensable to cartilage formation in chicken high density mesenchymal cell cultures (HDC) [C. Matta, J. Fodor, Z. Szijgyarto, T. Juhasz, P. Gergely, L. Csernoch, R. Zakany, Cytosolic free Ca(2+) concentration exhibits a characteristic temporal pattern during in vitro cartilage differentiation: a possible regulatory role of calcineurin in Ca-signalling of chondrogenic cells, Cell Calcium 44 (2008) 310-323]. Here, we report that chondrogenic cells secreted ATP and administration of ATP to the culture medium evoked Ca(2+) transients exclusively in the presence of extracellular Ca(2+) and only on day 3 of culturing, when the final commitment of chondroblasts occurs. Moreover, ATP caused elevated protein expression of the chondrogenic transcription factor Sox9 and stimulated cartilage matrix production. Expression pattern of different types of both ionotropic and metabotropic purinergic receptors was detected. Agonists of metabotropic receptors, ADP and UDP did not evoke any Ca(2+) transients and had no influence on cartilage formation, while UTP caused transient elevation of cytosolic Ca(2+) concentration in 3-day-old HDC without stimulating matrix production. Suramin, which blocks all P2X receptors but not P2X(4) did not impede the effects of ATP, furthermore, P2X(4) appeared in the plasma membrane fraction and gave signals with immunocytochemistry only from day 3. In summary, we suggest a role of ionotropic purinergic signalling of P2X(4) in the generation of ATP-dependent Ca(2+) transients of differentiating chondroblasts. PMID- 19297019 TI - Engraftment of human blood malignancies to the turkey embryo: a robust new in vivo model. AB - Xenografting of human blood malignancies to immunodeficient SCID mice is a powerful research tool. We evaluate here whether the immunodeficient turkey embryo can also serve as a xenograft host for human blood malignancies. Human leukemia, lymphoma and myeloma lines engrafted robustly into medullary and extramedullary tissues of turkey embryos as detected by PCR, FACS and histology in 8-10 days. Four of eleven patient AML samples also engrafted the bone marrow. Grafts of two lines responded to chemotherapy with doxorubicin. The turkey embryo therefore has the potential to be a complementary xenograft model for the study of human blood malignancies. PMID- 19297020 TI - Steroid effects on ZAP-70 and SYK in relation to apoptosis in poor prognosis chronic lymphocytic leukemia. AB - There is resurgent interest in glucocorticoids (GCs) in the treatment of poor prognosis chronic lymphocytic leukemia (CLL). Little is known however on how GCs induce apoptosis in CLL. Methylprednisolone (MP) induces apoptosis in ZAP-70 positive CLL more readily than in ZAP-70 negative CLL, which is in contrast to the effects of radiation and chemotherapy. The increased GC sensitivity of ZAP 70+ CLL was studied in relation to the expression status of ZAP-70 and the related signal transducing tyrosine kinase SYK. Both ZAP-70 and SYK were downregulated by GC treatment. Moreover, SYK was dephosphorylated by the phosphatase PTP1B of which the expression and translation levels were induced by GCs. Inhibition of PTP1B successfully restored ZAP-70 expression and SYK phosphorylation but did not interfere with GC-induced apoptosis. Therefore, the downregulation of ZAP-70 and P-SYK per se during treatment with GCs is not sufficient to induce apoptosis, and different mechanisms must therefore be responsible for the increased steroid sensitivity of ZAP-70+ CLL. PMID- 19297021 TI - Therapy-related acute myeloid leukemia after successful therapy for acute promyelocytic leukemia with t(15;17): A case report and literature review. PMID- 19297022 TI - The spectrum of phenotypes caused by variants in the CFH gene. AB - Complement factor H (CFH) is a complement inhibitor, which is present as a soluble protein and attached to cell surfaces throughout the human body. As such, CFH is a key player in complement homeostasis, inhibiting excessive activation of the complement cascade, with an emphasis on the alternative pathway. The significance of CFH is demonstrated by the broad range of phenotypes associated with specific CFH gene variants. This phenotypic spectrum includes renal phenotypes, such as membranoproliferative glomerulonephritis and atypical hemolytic uremic syndrome, as well as ocular phenotypes, such as basal laminar drusen and age-related macular degeneration. In addition, several overlapping phenotypes have been described in association with CFH gene variants. The phenotypic outcome of these CFH variants depends on their differential impact on plasma- and surface-bound CFH function. Consequently, distinct genotype-phenotype correlations may be observed. PMID- 19297023 TI - Promiscuity of MCMV immunoevasin of NKG2D: m138/fcr-1 down-modulates RAE-1epsilon in addition to MULT-1 and H60. AB - Both human and mouse cytomegalovirus (CMV) encode proteins that inhibit the activation of NK cells by down-regulating the cellular ligands for activating NK cell receptor, NKG2D. MCMV proteins m145, m152 and m155 interfere with the expression of all known NKG2D ligands, MULT-1, RAE-1 family members and H60, respectively, whereas m138 affects the expression of MULT-1 and H60. Here we show that m152 affects the maturation of newly synthesized RAE-1 molecules, but is not sufficient to prevent surface expression of RAE-1varepsilon. We have identified m138 as a main inhibitor of the surface expression of RAE-1varepsilon. In contrast to m152, m138 affects the surface-resident protein leading to its endocytosis, which can be prevented by a dynamin inhibitor. Moreover, we demonstrated that m138 does not need other viral proteins to down-modulate the expression of RAE-1varepsilon. PMID- 19297024 TI - CD59 but not DAF deficiency accelerates atherosclerosis in female ApoE knockout mice. AB - Although the complement system has been implicated in atherosclerosis, the influence of membrane-bound complement regulators in this process has not been well understood. We studied the role of two membrane complement regulators, decay accelerating factor (DAF) and CD59, in a murine model of atherosclerosis. DAF(-/ ) and CD59(-/-) mice were crossed with apolipoprotein E (ApoE)-deficient mice to generate DAF(-/-)ApoE(-/-) and CD59(-/-)ApoE(-/-) mice. Mice were fed a high fat diet (HFD) for 8 or 16 weeks. En face analysis showed that CD59 deficiency led to more extensive lesions in female ApoE(-/-) mice both at 8 weeks (2.07+/-0.27% vs.1.34+/-0.21%, P=0.06) and 16 weeks (17.13+/-1.14% vs. 9.72+/-1.14%, P<0.001). Similarly, lesions measured by aortic root sectioning were larger in female CD59( /-)ApoE(-/-) mice than in controls at 8 weeks of HFD feeding (20.74+/-1.33% vs. 13.12+/-1.46%, P<0.005). On the other hand, DAF deficiency did not significantly influence atherosclerosis in ApoE(-/-) mice. Immunohistochemistry revealed more abundant membrane attack complex (MAC) deposition and more collagen staining in the aortic roots of CD59(-/-)ApoE(-/-) mice. Unexpectedly, total plasma cholesterol levels in female CD59(-/-)ApoE(-/-) mice were found to be elevated compared with CD59(+/+)ApoE(-/-) mice. We conclude that CD59 but not DAF offered protection in atherosclerosis in the context of ApoE deficiency. The protective role of CD59 was gender-biased and most likely involved prevention of MAC mediated vascular injury, with possible contribution from an undefined effect on plasma cholesterol homeostasis. PMID- 19297025 TI - Validation of reference genes for quantitative measurement of immune gene expression in shrimp. AB - To accurately measure the relative expression of a target gene, mRNA expression data is routinely normalized with reference to an internal control gene. We examined the transcriptional stability of four internal control genes, beta actin, glyceraldehyde-3 phosphate dehydrogenase (GAPDH), elongation factor1-alpha (EF1-alpha), and 18S ribosomal RNA (18S rRNA) while measuring the mRNA expression of a gene encoding a pattern recognition protein, lipopolysaccharide and glucan binding protein (LGBP) gene, in healthy and white spot syndrome virus (WSSV) infected shrimp (Penaeus stylirostris) before and after (4, 8, 16 and 32 h) challenge using real-time RT-PCR. Here, we describe a method to rank the internal control genes based on a linear regression analysis. This analysis enables us to analyze the multivariate data set, e.g. time course study samples with control and treatment groups. Using the linear regression analysis and the WSSV challenged time course samples, GAPDH was found to be the most stable internal control gene followed by the genes EF1-alpha, 18S rRNA and beta-actin. Using the program geNorm, GAPDH was also found to be the most stable gene followed by the genes EF1-alpha, beta-actin and 18S rRNA. Using the program NormFinder, the ranking of the internal control genes were in the order of EF1-alpha>GAPDH>18S rRNA>beta-actin. The ability to compare the healthy and WSSV infected samples in parallel by the regression analysis makes this method a very useful approach while identifying the optimal reference gene for gene expression analysis. PMID- 19297026 TI - Cloning and characterization of SCART1, a novel scavenger receptor cysteine-rich type I transmembrane molecule. AB - We have cloned and characterized a novel murine transmembrane molecule, mSCART1 belonging to the scavenger receptor cysteine-rich superfamily. The cDNA encodes a polypeptide chain of 989 amino acids, organized as a type I transmembrane protein that contains eight extracellular SRCR domains followed by a transmembrane region and a cytoplasmic domain. The cytoplasmic domain contains two putative src kinase consensus substrate sequences, three additional potential phosphorylation sites, and two potential internalization motifs. Two possible secreted forms that lack the transmembrane region arise by alternative splicing. The murine SCART1 gene maps to chromosome 7 band F5 and the analysis of the genomic organization showed that the gene spans 12.86 kb and contains 14 exons. Quantitative real-time PCR analyses on murine tissues showed high mSCART1 mRNA expression in the lymph node, the trachea, and the lung, and low expression was found in the thymus, the spleen, the skin, and in tissues throughout the gastrointestinal tract. Comparative studies of the domain organization as well as the cytoplasmic domain of mSCART1 with the other members of the SRCR superfamily show that mSCART1 is highly related to the WC1 family of the SRCR superfamily. Finally, a novel human scavenger receptor cysteine-rich molecule with high homology to mSCART1 was identified by searching in the human genomic databases using the mSCART1 cDNA sequence. PMID- 19297027 TI - Manganese(III)-salens induce tumor selective apoptosis in human cells. AB - In order to explore the apoptotic and anti-tumor activities of metallo-salens, we synthesized several Mn(III)-salen derivatives (compds. 1-9) and analyzed their effects on cultured human cancer and non-cancer cells. Our results demonstrated that Mn(III)-salen derivatives affect cell viability, induce nuclear condensation and fragmentation in breast cancer cells (MCF7). Mn(III)-salen derivatives also induced caspase-3/7 activation and release of cytochrome-c from the mitochondria to cytosol suggesting that Mn(III)-salen derivatives induce apoptosis in human cells via mitochondrial pathway. Importantly, the nature of the substituent and the bridging spacer between diimino groups on the salen ligand play critical roles in determining the apoptotic activities of Mn(III)-salen derivatives. The IC(50) values for the active Mn(III)-salen derivatives lie within the range of 11 40microM in MCF7 cells. Most importantly, several Mn(III)-salen derivatives showed preferential cytotoxicity (2- to 5-fold) toward malignant breast cells (MCF7) over a non-malignant breast epithelial cell line (MCF10). Notably, the level of cytotoxicity and selectivity of the Mn(III)-salen derivatives towards MCF7 and MCF10 cells are very similar to cisplatin which indicate that Mn(III) salens are potential novel anti-tumor agent. PMID- 19297028 TI - Clinical impact of imipenem-resistant Pseudomonas aeruginosa bloodstream infections. AB - OBJECTIVES: To describe the incidence and clinical characteristics of imipenem resistant (IR) Pseudomonas aeruginosa bacteraemia. METHODS: We performed a retrospective study including all episodes of IR P. aeruginosa bacteraemia seen from January 2003 to December 2005 in a tertiary teaching hospital. RESULTS: There were 108 episodes of IR P. aeruginosa bacteraemia, which represented an incidence of 0.14 episodes per 1000 patient-days in 2003 and 0.11 episodes per 1000 patient-days in 2005. 83 of the episodes (77%) were nosocomially acquired. Most of patients had at least one underlying disease and had previously received antimicrobial treatment. The most frequent source was the urinary tract (31%), followed by unknown origin (22%). A total of 23 (21%) episodes were polymicrobial and 51 (47%) were caused by multidrug-resistant strains. The independent risk factors for mortality from IR P. aeruginosa bloodstream infection were a high risk source of the bacteraemia (OR: 4.6; 95% CI 1.7-12.4; p=0.01), and presentation with severe sepsis (OR: 2.8; 95% CI 1-7.8; p=0.05). CONCLUSIONS: Our study shows that the rates of IR P. aeruginosa bacteraemia remained stable throughout the study period. The source of bacteraemia and the clinical presentation with severe sepsis were the main determinants of the prognosis. PMID- 19297029 TI - Bovine immune response to papillomatous digital dermatitis (PDD)-associated spirochetes is skewed in isolate reactivity and subclass elicitation. AB - Papillomatous digital dermatitis (PDD) is a growing cause of lameness of dairy cattle worldwide. Farms with PDD-afflicted cows experience economic loss due to treatment costs, decreased milk production, lower reproductive efficiency and premature culling. Cows exhibit both humoral and cellular immune responses to PDD associated spirochetes. This study was undertaken to further characterize the bovine humoral response to PDD-associated spirochetes. Forty-seven sera samples collected from cattle (Field cattle) on three different dairy operations in Iowa were analyzed. In addition, sera were obtained from six young steers (Test cattle) that received a mixed inoculum of four previously isolated Treponema phagedenis-like spirochetes (1A, 3A, 4A and 5B) on two separate occasions. Relative levels of total IgG, IgG1, IgG2 and IgM reactive to each individual spirochete were determined. Field cattle had a higher mean antibody response to 5B compared to the other isolates and T. phagedenis. Test cattle reacted most strongly with 4A following initial exposure, shifting to a greater reactivity with 5B and a reactivity profile similar to field cattle following secondary exposure. No measurable IgM was detected. IgG1 was produced predominately in all cattle. Low to moderate levels of total IgG reactivity to T. phagedenis occurred with sera from all cattle. PMID- 19297030 TI - Canine neutrophil dysfunction caused by downregulation of beta2-integrin expression without mutation. AB - Canine leukocyte adhesion deficiency (CLAD) in Irish setters is caused by genetic defects of leukocyte integrin CD18 leading to recurrent bacterial infections. We report clinical features and analysis of neutrophil function from two mixed-breed canine littermates (one female and one male dog) similar to CLAD. The symptoms of pyogenic infection were first recognized at 3 months of age and since then the patients suffered from recurrent bacterial infections. These clinical findings were strongly suggestive of genetic phagocyte dysfunction. Neutrophil function tests revealed a marked reduction of serum-opsonized zymosan-mediated superoxide production in the two littermates. Neutrophils of the male dog revealed impaired integrin-mediated adherence and phagocytic activity, whereas ability of serum opsonization was normal. There was also a profound decrease of surface expression of CD11b/CD18 and beta2-integrin transcript level, detected by real-time RT-PCR without missense mutations unlike CLAD. Immunoblot analysis indicated that protein expression of cytochrome b(558) component gp91(phox), the cytosolic components p47(phox) and p67(phox) of NADPH oxidase components increased profoundly in the male. Our study suggests that decreased transcriptional levels of beta2-integrin without mutations, lead to downregulation of surface expression, resulting in multiple defects in adhesion-related neutrophil functions and consequently, recurrent bacterial infections from puppyhood. PMID- 19297031 TI - Parapharyngeal and retropharyngeal infections in children: argument for a trial of medical therapy and intraoral drainage for medical treatment failures. AB - OBJECTIVE: To review the outcomes of parapharyngeal infections (PPI) and retropharyngeal infections (RPI) in children after medical and surgical management. METHODS: The charts of all children who presented to a tertiary-care pediatric hospital between March 2005 and February 2007 with a diagnosis of parapharyngeal infection or retropharyngeal infection were reviewed. Thirty-two patient records were analyzed. RESULTS: The mean age of our patients was 4.4 years, 63% of whom were boys. The majority of patients presented during the winter months with some combination of fever, neck stiffness, and neck swelling. Ten patients had surgery less than 1 day after admission (early surgical intervention). Twenty-two patients were given a trial of at least 24h of intravenous (IV) antibiotics (medical trial group), nine of whom were treated exclusively with IV antibiotics (medical therapy alone). The remaining 13 patients failed medical therapy, and went on to surgical drainage (late surgical intervention). All patients requiring surgical drainage were treated successfully with an intraoral approach. Patients given a trial of IV antibiotics had an average hospital stay of 3.3 days (3.0 days for medical therapy alone and 3.5 days for late surgical intervention). Patients with early surgical intervention stayed 2.7 days. All patients were discharged home with oral antibiotics. There was no statistically significant difference in the length of stay between the medical trial and early intervention groups. Those who failed medical therapy had significantly larger abscesses than those who responded to medical therapy (1.53 versus 5.38 cm(2)). CONCLUSIONS: A trial of IV antibiotics does not adversely affect outcome and may obviate the need for surgery, particularly for patients with smaller abscesses. When surgery is required, an intraoral procedure is usually adequate. PMID- 19297032 TI - The effect of exogenous nitric oxide on alleviating herbicide damage in Chlorella vulgaris. AB - Herbicides present in the environment induce oxidative stress in plants. We investigated the roles of exogenous NO-regulated chlorophyll synthesis, antioxidant enzyme activity and gene expression in herbicide-treated unicellular green algae Chlorella vulgaris. Atrazine (100 microg/L) or glufosinate (10 mg/L) treatment alone or in combination with 20 microM sodium nitroprussiate (SNP, 10 microM with glufosinate) was administered to algae for a short time period of 48 h to observe changes in physiology and gene transcription and expression. Supplementation of atrazine or glufosinate with low SNP significantly reduced H2O2, reactive oxygen species (ROS) and malondialdehyde (MDA) induction by herbicides. Supplementation also increased chlorophyll content and antioxidant enzymes, superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activity, as compared to herbicide treatment alone. This trend suggests an effect of NO on the scavenging of ROS. Furthermore, the expression of photosynthesis genes (psbC, psaB, chlB and rbcL) was also upregulated by supplementation of low SNP, thus maintaining the normal photosynthetic function. However, high concentration of SNP (100 microM) in combination with herbicides aggravated damage to algae, including increases in H2O2, ROS and MDA and decreases in chlorophyll content, antioxidant enzyme activity and photosynthesis genes transcription. 2-Phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (PTIO), the NO scavenger, was also examined in this study; the results showed that PTIO could neutralise the effect of low SNP. Data also showed that an exogenous supply of NO protects Chlorella vulgaris against the toxicity of herbicides by protecting against oxidant substances and increasing the transcription of related genes. PMID- 19297033 TI - Effects of the cyanobacterial neurotoxin beta-N-methylamino-L-alanine on the early-life stage development of zebrafish (Danio rerio). AB - beta-N-Methylamino-L-alanine (BMAA), a neurotoxic amino acid, is produced by members of all known groups of cyanobacteria. In the presence of added carbonate, BMAA generates an analogue of glutamate which has been associated with motor neuron (MN) diseases via a mechanism of motor neurone specific excitotoxicity. The toxicity of BMAA has been established in various mammalian test models, but the widespread aquatic production of BMAA raises questions of BMAA toxicity to aquatic organisms. Zebrafish (Danio rerio) embryos were exposed to varying concentrations of BMAA (5-50,000 microgl(-1)) with and without added carbonate. BMAA exposure induced a range of neuro-muscular and developmental abnormalities in D. rerio, which can be directly related to disruptions to glutamatergic signalling pathways. When exposed to BMAA plus added carbonate, the incidence of pericardial oedema increased by up to 21% in test subjects, correlating with a reduction in heart rate. Increased incidence of abnormal spinal axis formation was seen in all D. rerio larvae exposed to BMAA concentrations of >or=50microgl( 1), with a further 10% increase from >or=500 microgl(-1) BMAA when carbonate species were present. A dose-dependent increase in clonus-like convulsions was observable in embryos exposed to >or=5 microgl(-1) BMAA+/-added carbonate. This is the first study on the neuro-muscular and developmental effects of BMAA exposure on aquatic vertebrates. The present findings, plus the potentially widespread production of BMAA in aquatic cyanobacteria, indicate a need for information of exposure levels, duration and toxic outcomes in aquatic biota. PMID- 19297034 TI - Metal stress induces programmed cell death in aquatic fungi. AB - Aquatic hyphomycetes are a group of fungi that play a key role in organic matter turnover in both clean and metal-polluted streams. We examined the ability of Cu or Zn to induce programmed cell death (PCD) in three aquatic hyphomycete species through the evaluation of typical apoptotic markers, namely reactive oxygen species (ROS) accumulation, caspase-like activity, nuclear morphological alterations, and the occurrence of DNA strand breaks assessed by TUNEL assay. The exposure to both metals induced apoptotic events in all tested aquatic fungi. The most tolerant fungi either to Zn (Varicosporium elodeae) or Cu (Heliscussubmersus) exhibited higher levels of PCD markers, suggesting that PCD processes might be linked to fungal resistance/tolerance to metal stress. Moreover, different patterns of apoptotic markers were found, namely a PCD process independent of ROS accumulation in V. elodeae exposed to Cu, or independent of caspase-like activity in Flagellospora curta exposed to Zn, or even without the occurrence of DNA strand breaks in F. curta exposed to Cu. This suggests that a multiplicity of PCD pathways might be operating in aquatic hyphomycetes. The occurrence of a tightly regulated cell death pathway, such as PCD, in aquatic hyphomycetes under metal stress might be a part of the mechanisms underlying fungal acclimation in metal-polluted streams, because it would allow the rapid removal of unwanted or damaged cells sparing nutrients and space for the fittest ones. PMID- 19297035 TI - Urinary incontinence and behavioral symptoms are independent risk factors for recurrent and injurious falls, respectively, among residents in long-term care facilities. AB - Numerous risk factors of falls, including urinary incontinence and behavioral symptoms have been identified among elderly people in long-term care settings. However, it remains uncertain whether incontinence or behavioral symptoms are associated with recurrent falls and injurious falls. The purpose of this research was to examine the association between various types of falls and urinary incontinence or behavioral symptoms among the residents of long-term care facilities using the Cox proportional hazards models. The participants were 1082 older people (327 men and 755 women) who were admitted to facilities between 1 April 2003 and 31 March 2004. Fall experience, urinary incontinence, and behavioral symptoms were followed for up to 6 months or until death or discharge. The functional status, comorbidity, and prescribed medications were determined at the baseline. Multivariate analysis revealed that urinary incontinence and behavioral symptoms were independent risk factors of falls during the follow-up period. However, urinary incontinence was a risk factor for recurrent falls but not for injurious falls. In contrast, behavioral symptoms were an independent risk factor for injurious but not for recurrent falls. The results suggested that treatment or management of urinary incontinence and behavioral symptoms should be considered to prevent falls in long-term care settings. PMID- 19297036 TI - Exclusion of Cx43 gene mutation as a major cause of criss-cross heart anomaly in man. AB - Criss-cross heart is a rare congenital cardiac defect characterized by crossing of the atrioventricular valves and of the inflow streams due to the twisting of the ventricles about their long axis. The aetiology of criss-cross heart has not been understood yet. Mice homozygous for Cx43 deficiency show a delay in normal looping of ascending limb of the heart tube, which temporarily retains a more symmetric middle position. Persistence of this condition results in a "criss cross" configuration, with the atrioventricular cushions rotated 90 degrees , a horizontal muscular ventricular septum, and a parallel course of the endocardial ridges of the outflow tract. We screened the entire coding region of the Cx43 gene in a group of well characterized patients with criss-cross heart, to evaluate whether Cx43 gene mutations cause criss-cross heart in humans. No pathogenic mutation was identified, suggesting that Cx43 mutations are not responsible for criss-cross heart in humans or are not a major cause for this defect. PMID- 19297037 TI - Coronary slow flow: is it a disease or normal variant? AB - The exact pathophysiological mechanism of coronary slow flow remains unclear although some possible mechanisms such as altered microvascular tone, diffuse vascular pathology, and acute release of coronary vasoconstrictor mediators have been proposed. Therefore we need new studies regarding responsible etiologies to answer the question 'Is coronary slow flow a disease or normal variant?' PMID- 19297038 TI - Is diabetes the cost to pay for a greater cardiovascular prevention? AB - The recent JUPITER (Justification for the Use of statins in Primary prevention: an Intervention Trial Evaluating Rosuvastatin) trial is another study providing evidence about the effectiveness of statin therapy in reducing cardiovascular risk. Yet, in this study significantly higher glycated hemoglobin levels and incidence rates of diabetes were observed in persons treated with rosuvastatin than the placebo group. It should be noted that adverse effects on glucose metabolism have already been reported, albeit rarely, in previous trials with statins. Although the exact mechanisms involved are unknown, it seems that statins may deteriorate glycemic control by decreasing different metabolites, including isoprenoid and ubiquinone, normally produced during the process of cholesterol synthesis. We therefore suggest that, if statins are prescribed, patients should be monitored closely for blood glucose control even though the higher incidence of diabetes by statin therapy may represent a rare finding. PMID- 19297039 TI - CORD: COmparison of Recommended Doses of ace inhibitors and angiotensin II receptor blockers. AB - The CORD trials tested ramipril and losartan in patients with hypertension. CORD A randomised 4016 patients with blood pressure (BP) <160/100 mm Hg, who had been treated with an ACEI for >3 months. The patients discontinued ACEI and switched to losartan. After 1 month the BP decreased to 7.7/4.7 mm Hg (p<0.001) and after 1 year to 13.8/8.7 mm Hg (p<0.001). CORD B compared ramipril and losartan in 3813 patients with hypertension who were not being treated with an ACEI or ARB. The patients were randomised to ramipril (n=1926) or losartan (n=1887). After 1 year the BP decreased in the ramipril group to 21.8/13.7 mm Hg (p<0.001) and in the losartan group to 22.0/13.3 mm Hg (p<0.001). No significant differences were found between the groups. No differences were in serious adverse events. Dry cough was more frequently after ramipril (33 vs 4, p<0.001). PMID- 19297040 TI - Correlation of NT-proBNP with metabolic liver function as assessed with (13)C methacetin breath test in patients with acute decompensated heart failure. AB - Metabolic liver function is significantly decreased in patients with advanced chronic heart failure (CHF). The purpose of the study was to evaluate correlation of NT-proBNP with metabolic liver function as assessed with (13)C-methacetin breath test in patients with acute decompensation of chronic heart failure (ADHF). NT-proBNP, (13)C-methacetin breath test and investigation of other biochemical parameters were realized on days 0-2 (V1) and 2-7 (V2) after hospital admission in 13 consecutive ADHF patients. NT-proBNP and metabolic liver function as assessed with (13)C-methacetin breath test didn't change in the first days after hospital admission in ADHF patients. Correlation of NT-proBNP with the degree of metabolic liver function impairement was not significant. The study revealed significant correlation of NT-proBNP with serum ceruloplasmin and significant inverse correlation of liver function with total iron binding capacity in patients with ADHF. The results of this pilot study should be confirmed in a study with larger patient population. PMID- 19297041 TI - Comments on "Asymptomatic hypoxia in a young pregnant lady--unusual presentation of atrial septal defect", Krishnamoorthy S, et al. AB - In their recently published article, Krishnamoorthy and coworkers describe a case of an asymptomatic hypoxia due to atrial septal defect in a pregnant woman. However, before accepting their conclusion, some issues require further clarification: (a) What were the patient's haemodynamic findings in cath-angio before the closure of ASD? (b) What was the final arterial saturation after device closure of the ASD? PMID- 19297043 TI - Usefulness of left atrial volume for the diagnosis of diastolic heart failure: an echocardiographic-catheterization study. AB - The present study attempted to determine the accuracy of left atrial volume (LAVi) by transthoracic echocardiography in the diagnosis of diastolic heart failure (DHF) in patients presenting with chronic, isolated dyspnea. We included 28 consecutive patients with a left ventricular ejection fraction >50% without prior history of heart failure. DHF was authenticated in 20 patients by invasive left ventricular end-diastolic pressure >16 mmHg. By logistic regression analysis, LAVi was predictive of DHF (p=0.015). LAVi>38 ml/m(2) was a useful predictor of DHF (area under the ROC curve of 0.84 [0.65-0.95], p<0.001, sensitivity 60%, specificity 100%). The standard cut-off value of 34 ml/m(2) was 70% sensitive and 88% specific. PMID- 19297044 TI - A nation-wide epidemiological study of acute bovine respiratory disease in France. AB - Acute bovine respiratory disease (ABRD) is a respiratory syndrome caused by various infectious agents, and represents a major economic and health problem in cattle herds worldwide. The aim of the study was to present how we can describe the epidemiological characteristics of ABRD in France, including risk factor analysis and spatio-temporal pattern investigation. The data were collected by specialized surveyors during the 2001 animal survey conducted by the French Ministry for Agriculture and Fisheries, from a representative sample of cattle herds. The health questionnaire concerned farm characteristics, practice of BRD vaccination, presence of ABRD, time of occurrence and number of affected animals for the period November 2000-October 2001. After a descriptive analysis of the syndrome, a space-time scan statistic was performed to detect potential clusters, and a logistic regression model was used for analysis of risk factors. The study sample included 16,581 cattle herds as representative of the 260,000 French herds. The ABRD cumulative incidence at farm level was 9.8%, the cumulative incidence at animal level was 2.1%, the cumulative mortality 0.1% and the lethality proportion 6.5%. ABRD occurred mainly during cold months with an epidemic peak in December. Spatial repartition of the syndrome showed that the whole territory was affected, with a higher number of cases in the north-east region, the main bovine production area in France. Three space-time clusters were identified in the period November 2000-February 2001. ABRD was significantly associated with production type, herd size, and BRD vaccination. Beef herds were less affected than dairy herds, and increased herd size increased the risk of ABRD. The outbreak was clustered in space and time, suggesting a common infectious agent for the epidemic in the detected areas. The influence of production type and herd size on syndrome occurrence highlighted the importance of management practices, human movement and animal density. PMID- 19297045 TI - Bayesian estimation of variance partition coefficients adjusted for imperfect test sensitivity and specificity. AB - The variance partition coefficient (VPC) measures the clustering of infection/disease among individuals with a specific covariate pattern. Covariate pattern-specific VPCs provide insight to the groups of individuals that exhibit great heterogeneity and should be targeted for intervention. VPCs should be taken into consideration when planning study designs, modeling data and estimating sample sizes. We present a Bayesian discrete mixed model for the estimation of covariate-pattern-specific VPCs when measurement of the infection/disease is based on an imperfect test. The utility of the presented model is demonstrated with three applications. In all cases, imperfect tests biased VPC estimates towards the null but corrected estimates could be obtained by modeling the sensitivity and specificity of the test procedure with beta distributions. The comparison of adjusted VPCs between the intercept only and the fitted models with higher level covariates explained the portion of heterogeneity in the data that was accounted for by the covariates. PMID- 19297046 TI - Heifer rearing conditions affect length of productive life in Swedish dairy cows. AB - Associations between rearing conditions and the risk of culling in dairy cows were studied by survival analysis. Data were collected from 1039 Swedish Red cows, 1029 Swedish Holsteins, and 56 cows of other milk or cross-breeds, representing all female animals born in 109 Swedish herds during 1998. Length of productive life was defined as the number of days from 1st calving to culling. The applied Weibull proportional hazards model included time-independent effects of breed, housing from 3 to 7 months of age, number of housing changes before calving, grazing before 1st calving, herd median age at 1st calving, age at 1st calving, cow housing, herd lactational incidence risk of veterinary-reported clinical mastitis, and the random effect of herd. Time-dependent effects were year, month, the interaction year by month, parity, number of breedings, pregnancy status, the interaction parity by pregnancy status, herd mean milk production level, relative milk yield within breed-parity, and veterinary reported clinical mastitis. The lactation was divided into six stages in which pregnancy status was assumed to be known by the farmer and culling could occur. Median productive life time in culled cows was 780 d and 14% of the records were censored due to terminated data collection. An individual calving age of 28.2 30.9 months was associated with the highest culling risk, 1.2-fold higher than calving at < or = 25.3 months, whereas the risk decreased almost linearly with a higher herd median age at 1st calving. Housing in slatted pens with >7 calves from 3 to 7 months was associated with a 1.7-fold increase in risk, relative to litter pens. If a cow had changed housing system 4 times before 1st calving it increased the risk of culling 1.4 times, relative to two housing changes. These results show that rearing factors affect the productive life time of dairy cows in Swedish family operations. PMID- 19297047 TI - Sole ulcers in Finnish dairy cattle. AB - The Finnish Healthy Hooves project was set up to determine the frequency of, and risk factors for various hoof lesions in Finnish dairy herds. Data were collected in the years 2003 and 2004. A large dataset of over 74,000 cow-level observations recorded by hoof trimmers was merged with production data from the Finnish Agricultural Data Processing Centre Ltd. Ultimately, data from a single lactation from each of 16,792 cows in 703 herds were used for the analyses in this paper. Three-level hierarchical logistic models with hoof trimmer and farms (within hoof trimmer) as random effects were fit to data sets of tie stall (TS) and loose housing (LH) herds separately. The outcome of interest was the presence or absence of a sole ulcer in one or more legs of a cow during the lactation of interest. Cows examined once had a risk of sole ulcer 5.23% in tie stall herds and 7.58% in LH herds. As the number of examinations increased the odds of a diagnosis of sole ulcer increased substantially (2 and 3+ examinations had odds ratios (ORs) of 1.42 and 3.42 in TS herds and 2.77 and 6.89 in LH herds). Breed had a large effect on the risk of sole ulcer with Holsteins 2.89 times more likely to be affected than Ayrshires in TS herds and 2.94 times in LH herds. In TS herds, the presence of other hoof lesions such as haemorrhages (OR = 2.97), heel-horn erosions (OR = 2.10) and corkscrew claw (OR = 2.83) increased the risk of a sole ulcer developing. In LH herds, only haemorrhages (OR = 1.80) were a significant risk factor when parity was > or = 2. In TS herds, use of mats (compared to hard flooring) significantly reduced the risk of sole ulcers (OR = 0.49). The effect of parity on the risk of sole ulcer was greatest when parity > or = 4 but this effect was only significant in tie stalls (OR = 1.86). When analyses were restricted to cows with parity > or = 2, similar results were obtained for the risk factors identified above. In addition, parity became highly significant in TS and LH (OR 2.31 and 2.23, respectively when parity was 4+). In TS herds, herd average milk production was significantly associated with a decrease risk of sole ulcer (OR = 1.28 per 1000 kg decrease) but there was no effect of production at the cow level (measured as deviation from the herd mean). No significant effects of production were observed in LH herds. PMID- 19297048 TI - Stereotactic radiotherapy with real-time tumor tracking for non-small cell lung cancer: clinical outcome. AB - PURPOSE: To report the clinical outcome of treatment using real-time tumor tracking for 70 patients with inoperable stage I non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: Seventy inoperable patients with peripherally located early-stage NSCLC were treated with 45 or 60 Gy in three fractions using CyberKnife. Pathology was available in 51% of patients. Thirty-nine patients had a T1-tumor and 31 had a T2-tumor. Markers were placed using the vascular, percutaneous intra-, or extra-pulmonary approach, depending on the risk of pneumothorax. RESULTS: The actuarial 2-year local control rate for patients treated with 60 Gy was 96%, compared to 78% for patients treated with a total dose of 45 Gy (p=0.197). All local recurrences (n=4) occurred in patients with T2 tumors. Overall survival for the whole group at two years was 62% and the cause specific survival was 85%. The median follow-up was 15 months. Grade 3 toxicity occurred in two patients (3%) after marker placement. Treatment-related late grade 3 toxicity occurred in 7 patients (10%). No grade > or = 4 toxicity occurred. CONCLUSION: Excellent local control of 96% at 1- and 2-years was achieved using 60 Gy in three fractions for NSCLC patients treated with the real time tumor tracking. Toxicity was low. PMID- 19297049 TI - High-dose radiotherapy or concurrent chemo-radiation in lung cancer patients only induces a temporary, reversible decline in QoL. AB - BACKGROUND AND PURPOSE: Aggressive radiotherapy or concurrent chemo-radiation therapy for lung cancer leads to a high incidence of severe, mostly esophageal, toxicity. The purpose of this study was to investigate the evolution of quality of life (QoL) in patients with lung cancer, selected for curative radiotherapy (RT) or chemo-RT. METHODS: Seventy-five lung cancer patients completed a longitudinal the EORTC QLQ-C30 and LC13. Linear mixed regression models were fitted to investigate the impact of different factors on overall QoL. RESULTS: Overall QoL decreased shortly after the end of RT (4 points, p=0.19), but increased back to baseline within 3months. Mean scores of role functioning (p=0.018), cognitive functioning (p=0.002), dyspnoea (EORTC QLQ-LC13; p=0.043), dysphagia (p=0.005) and hoarseness (p=0.029), showed a significant worsening over time. Emotional functioning (p=0.033) improved significantly over time. Severe esophagitis (> or = grade 2) was reported in only 12% of the patients. Next to maximal esophageal toxicity > or = grade 2 (p=.0.010), also tumor stage IIIA (p<0.001), tumor stage IIIB (p=0.003), gender (p=0.042) and fatigue (p<0.001) appeared to be significant predictors of QoL. CONCLUSION: High-dose radiotherapy or concurrent chemo-radiation in the treatment of lung cancer seems to be a well tolerated treatment option with preservation of QoL. PMID- 19297050 TI - Preoperative radiotherapy and local excision of rectal cancer with immediate radical re-operation for poor responders. AB - BACKGROUND AND PURPOSE: To report an early analysis of prospective study exploring preoperative radiotherapy and local excision in rectal cancer. MATERIALS AND METHODS: Mucosa at tumour edges was tattooed. Patients with cT1-3N0 tumour <3-4 cm were treated with either 5x5Gy+4Gy boost (N=31) or chemoradiation (50.4Gy+5.4Gy boost, 1.8Gy per fraction+5-fluorouracyl and leucovorin; N=13). Thirteen patients from the short-course group were unfit for chemotherapy. The interval from radiation to full-thickness local excision was 6 weeks. The protocol called for conversion to a transabdominal surgery in case of ypT2-3 disease or positive margin. RESULTS: The postoperative complications requiring hospitalization were recorded in 9% of patients. The rate of pathological complete response was 41%. The rate of patients requiring conversion was 34%; however, 18% actually underwent conversion and the remaining 16% refused or were unfit. During the 14 months of median follow-up, local recurrence was detected in 7% of patients and all underwent salvage surgery. Of 19 patients in whom initially anterior resection was likely, 16% had abdominoperineal resection performed for a conversion or as a rescue procedure. CONCLUSION: Our study suggests that the short-course radiation prior to local excision is a treatment option for high-risk patients. PMID- 19297051 TI - The effect of different lung densities on the accuracy of various radiotherapy dose calculation methods: implications for tumour coverage. AB - PURPOSE: To evaluate against Monte-Carlo the performance of various dose calculations algorithms regarding lung tumour coverage in stereotactic body radiotherapy (SBRT) conditions. MATERIALS AND METHODS: Dose distributions in virtual lung phantoms have been calculated using four commercial Treatment Planning System (TPS) algorithms and one Monte Carlo (MC) system (EGSnrc). We compared the performance of the algorithms in calculating the target dose for different degrees of lung inflation. The phantoms had a cubic 'body' and 'lung' and a central 2-cm diameter spherical 'tumour' (the body and tumour have unit density). The lung tissue was assigned five densities (rho(lung)): 0.01, 0.1, 0.2, 0.4 and 1g/cm(3). Four-field treatment plans were calculated with 6- and 18 MV narrow beams for each value of rho(lung). We considered the Pencil Beam Convolution (PBC(Ecl)) and the Analytical Anisotropic Algorithm (AAA(Ecl)) from Varian Eclipse and the Pencil Beam Convolution (PBC(OMP)) and the Collapsed Cone Convolution (CCC(OMP)) algorithms from Oncentra MasterPlan. RESULTS: When changing rho(lung) from 0.4 to 0.1g/cm(3), the MC median target dose decreased from 89.2% to 74.9% for 6 MV and from 83.3% to 61.6% for 18 MV (of dose maximum in the homogenous case at both energies), while for both PB algorithms the median target dose was virtually independent of lung density. CONCLUSIONS: Both PB algorithms overestimated the target dose, the overestimation increasing as rho(lung) decreased. Concerning target dose, the AAA(Ecl) and CCC(OMP) algorithms appear to be adequate alternatives to MC. PMID- 19297052 TI - Growth and survival at chiller temperatures of Arcobacter butzleri. AB - Arcobacter butzleri is prevalent on chicken products. Arcobacter spp. are generally isolated in only low numbers from the chicken gut, so chicken carcasses may be contaminated by A. butzleri that proliferate in the slaughterhouse environment. To address this issue, we examined the behaviour of A. butzleri ATCC 49616 and newly isolated A. butzleri strains under conditions likely to prevail in the slaughterhouse environment using a chicken meat juice medium (CMJ). CMJ supported growth of A. butzleri at 15 degrees C, the recognised minimal growth temperature of this organism, and at 10 degrees C. At 5 degrees C, CMJ enhanced survival of A. butzleri as compared with survival in Brain Heart Infusion with less than a one log reduction after 77 days incubation. Lastly, we examined the ability of A. butzleri to form biofilms and found that the organism produces biofilm at temperatures ranging from 5 to 37 degrees C. Given the ability to survive, multiply and form biofilm under chilled conditions A. butzleri appears well suited for establishment in food processing and slaughterhouse environments. PMID- 19297053 TI - RAPGEF1 gene variants associated with type 2 diabetes in the Korean population. AB - Under the activation of insulin receptors, glucose transporter 4 (Glut4) translocation is regulated by two signal transduction pathways. These pathways are the PI 3-kinase-dependent pathway and the CAP/TC10 pathway. The adaptor protein Rap guanine exchange factor 1 (RAPGEF1) also known as C3G is a component of the CAP/TC10 pathway. Defects in the RAPGEF1 protein may contribute to insulin resistance and type 2 diabetes. Recently, the RAPGEF1 gene was suggested to be involved in the development of type 2 diabetes by FUSION study. To investigate this association in the Korean population, we sequenced the RAPGEF1 gene in 24 unrelated individuals and identified 39 sequence variants. Eleven single nucleotide polymorphisms (SNPs) were selected and genotyped in 1122 Korean patients with type 2 diabetes. There were 1138 non-diabetic controls. Using a logistic regression analysis, a significant association was found between SNP rs11243444 in the RAPGEF1 gene and type 2 diabetes [OR=0.490 (95% CI 0.296 0.813), p=0.006] in the recessive model, leading the protective effect of the GG genotype on the disease development. The present study examines genetic polymorphisms in the RAPGEF1 gene, and the positive association between one polymorphism and type 2 diabetes in the Korean population. PMID- 19297054 TI - Waist circumference reduction after insulin detemir therapy in type 2 diabetes patients previously treated with NPH. AB - We studied the weight-sparing effect and treatment satisfaction when switching from NPH to insulin detemir in type 2 diabetes. Mean HbA(1c) (P<0.05) and waist circumference (P<0.05) were reduced while treatment satisfaction improved (P<0.03). No weight gain was observed. Detemir improves glycemic control, treatment satisfaction, and may provide additional weight-sparing benefits. PMID- 19297055 TI - Membrane cholesterol enrichment prevents Abeta-induced oxidative stress in Alzheimer's fibroblasts. AB - A growing body of evidence implicates low membrane cholesterol in the pathogenesis of Alzheimer's disease (AD). Here we show that Abeta42 soluble oligomers accumulate more slowly and in reduced amount at the plasma membranes of PS-1L392V and APPV717I fibroblasts from familial AD (FAD) patients enriched in cholesterol content than at the counterpart membranes. The Abeta42-induced production of reactive oxygen species (ROS) and the increase in membrane lipoperoxidation were also prevented by high membrane cholesterol, thus resulting in a higher resistance to amyloid toxicity with respect to control fibroblasts. On the other hand, the recruitment of amyloid assemblies to the plasma membrane of cholesterol-depleted fibroblasts was significantly increased, thus triggering an earlier and sharper production of ROS and a higher membrane oxidative injury. These results identify membrane cholesterol as being key to Abeta42 oligomer accumulation at the cell surfaces and to the following Abeta42-induced cell death in AD neurons. PMID- 19297056 TI - An elemental diet fed, enteral or parenteral, does not support growth in young pigs with exocrine pancreatic insufficiency. AB - BACKGROUND & AIMS: Young individuals with exocrine pancreatic insufficiency (EPI) show growth reduction that can be reversed by dietary pancreatic enzyme supplementation. Here we investigated whether feeding an elemental diet could replace the growth-promoting effect of enzyme supplementation in EPI pigs. METHODS: Weaned pigs with intact pancreas (control) or pancreatic duct-ligated (EPI pigs) were given a commercial pig feed, a fat-enriched diet, or an elemental diet, intragastrically and intravenously, with or without porcine pancreatin (Creon) supplementation for 1 week. RESULTS: Control pigs, irrespective of receiving pig feed or an elemental diet, increased their body weight by 13.4 20.1%, while EPI pigs showed negligible weight gain. Giving a fat-enriched diet did not improve growth of the EPI pigs. However, if the EPI pigs were supplemented with pancreatin in combination with fat-enriched feed or the elemental diet, i.v., their body weight increased by 16.6 %and 8.5%, respectively. CONCLUSION: Control pigs maintained normal growth, independently of the diet being given in polymeric or elemental form, while EPI pigs showed impaired growth when receiving the same diets without enzyme supplementation. Pancreatic juice and enzyme preparations, in addition to their digestive properties, also appear to affect nutrient assimilation and anabolism in young individuals. PMID- 19297057 TI - Two non-surgical treatments for chondrodermatitis nodularis helicis. PMID- 19297058 TI - Technical notes published in BJOMS over a 2-year period--should we be doing it differently? AB - Between January 2007 and December 2008, 44 technical notes or related publications (such as letters) were published in the British Journal of Oral and Maxillofacial Surgery (BJOMS). These covered most of the remit of the specialty and ranged from operative surgical techniques to the use of digital photographs to orientate surgical specimens. However, there would seem to be very little feedback on the value of these articles in everyday practice. We reviewed these technical notes and assessed readability, the value and use of illustrations, the possible expense and/or equipment required in their use, and finally the frequency that the techniques could be used. The anonymised publications were read and scored by a minimum of two dentally qualified senior house officers, two doubly qualified specialist registrars, an SAS grade and two consultants in oral and maxillofacial surgery. The six techniques that gained the highest mean average score are briefly discussed. Although we used a relatively small number of assessors who might not be representative of the whole BJOMS readership, this study would suggest that some sort of change in the way that these technical notes are published should be considered. Options might include inviting a commentary from the reviewers who have tried the technique and also encouraging colleagues to report their experiences of these techniques in the 'letters to the editor' section. PMID- 19297060 TI - An unusual delayed complication of inferior alveolar nerve block. AB - Systemic and localised complications after administration of local anaesthetic for dental procedures are well recognised. We present two cases of patients with trismus and sensory deficit that arose during resolution of trismus as a delayed complication of inferior alveolar nerve block. PMID- 19297061 TI - Genotoxic effect of polycyclic aromatic hydrocarbons in the metropolitan area of Porto Alegre, Brazil, evaluated by Helix aspersa (Muller, 1774). AB - The purpose of this study was to biomonitor metropolitan areas of Porto Alegre (Brazil) for PAHs associated with atmospheric particles and check their effects on the DNA of the land mollusk Helix aspersa. The sampling sites are located in an urban area with heavy traffic: (i) Canoas, (ii) Sapucaia do Sul, and (iii) FIERGS/Porto Alegre. The samples were collected during a continuous period of 24 hours during 15 days using Stacked Filter Units (SFU) on polycarbonate filters (two separated size fractions: PM(10-2.5) and PM(< 2.5)). The concentrations of 16 major PAHs were determined according to EPA. Comet assay on H. aspersa hemolymph cells was chosen for genotoxicity evaluation. This evaluation shows that, in general, the smaller PM-size fractions (PM(< 2.5)) have the highest genotoxicity and contain higher concentrations of extractable organic matter. In addition, associations between chemical characteristics and PM carcinogenicity tend to be stronger for the smaller PM-size fractions. PMID- 19297062 TI - Are Bavarian forests (southern Germany) at risk from ground-level ozone? Assessment using exposure and flux based ozone indices. AB - Exposure and flux-based indices of O3 risk were compared, at 19 forest locations across Bavaria in southern Germany from 2002 to 2005; leaf symptoms on mature beech trees found at these locations were also examined for O3 injury. O3 flux modelling was performed using continuously recorded O3 concentrations in combination with meteorological and soil moisture data collected from Level II forest sites. O3 measurements at nearby rural open-field sites proved appropriate as surrogates in cases where O3 data were lacking at forest sites (with altitude dependent average differences of about 10% between O3 concentrations). Operational thresholds of biomass loss for both O3 indices were exceeded at the majority of the forest locations, suggesting similar risk under long-term average climate conditions. However, exposure-based indices estimated higher O3 risk during dry years as compared to the flux-based approach. In comparison, minor O3 like leaf injury symptoms were detected only at a few of the forest sites investigated. Relationships between flux-based risk thresholds and tree response need to be established for mature forest stands for validation of predicted growth reductions under the prevailing O3 regimes. PMID- 19297063 TI - Overweight status, self-perception, and suicidal behaviors among adolescents. AB - Suicide is the third leading cause of death among adolescents in the USA. The suicide rate for individuals 15-19 years of age, while having declined since the early 1990s, has recently shown signs of an increasing trend. The prevalence of being overweight has also steadily risen among adolescents, and has tripled since 1960. This study utilizes data from the Youth Risk Behavioral Surveillance System (1999-2007) to explore the relationship between the perception of being overweight and suicidal behaviors. Studies have shown a high degree of correlation between overweight status, depressive disorders, and suicidal behaviors. This study analyzes these indicators in conjunction with individuals' perception of their weight. The empirical methodology is based on simultaneous equations models and stratified samples to gauge whether the link between overweight indicators and suicide is causal or whether it is driven by other factors. Results indicate that body dissatisfaction, as measured by the perception of being overweight, has a strong impact on all suicidal behaviors for girls. It raises the risk of suicide ideation by 6.1 percentage points, suicide attempt by 3.6 percentage points, and a serious suicide attempt by 0.5 percentage points. Results are generally insignificant for males. Conditional on overweight perception, actual weight does not generally have an independent effect on suicidal behaviors. Policies aimed at reducing the prevalence of overweight among adolescents may further reduce suicidal behaviors by limiting overweight perception, especially among girls. However, the independent role of perception also highlights the importance of educating youths and fostering healthy attitudes regarding body image. PMID- 19297064 TI - Extending specialist palliative care to people with heart failure: semantic, historical and practical limitations to policy guidelines. AB - This paper explores the continuities and discontinuities in recent policy on the extension of palliative care to people with heart failure in the UK. It focuses on how professionals in cardiology and specialist palliative care negotiate their disciplinary boundaries within the context of these policy moves. It draws out the semantic, historical and practical tensions between the core values of cardiology, with its focus on 'living with heart failure', and specialist palliative medicine, with its focus on 'dying with heart failure'. A focus on negotiation of interdisciplinary boundaries reveals different engagements with notions of dying and palliation rather than simply different disease trajectories. Further, uncertainty about prognosis and the probability of sudden death pose a challenge to two core principles of specialist palliative care: 'open awareness' and 'good death'. We are not suggesting that these differences are insurmountable. Rather, in highlighting these tensions, our aim is to problematise the relationship between policy and practice, as being mediated by negotiations of disciplinary values (culture) within a local context. We conclude by recommending a wider discussion on notions of 'open awareness' and how professionals within different medical disciplines engage with alternate ways of dealing with uncertainty and sudden death as part of human condition. The arguments presented here are based on data and analysis from a larger qualitative study conducted during 2004-2005 in north and central England. PMID- 19297065 TI - The effects of exercise interventions on quality of life in clinical and healthy populations; a meta-analysis. AB - The aim of the study was to provide an overview of the effect of exercise interventions on subjective quality of life (QoL) across adult clinical populations and well people, and to systematically investigate the impact of the exercise setting, intensity and type on these outcomes. From a systematic search of six electronic databases, 56 original studies were extracted, reporting on 7937 sick and well people. A meta-analysis was conducted on change in QoL from pre- to post-intervention compared with outcomes from a no-exercise control group, using weighted (by the study's sample size) pooled mean effect sizes and a fixed-effects model. Significant differences in outcome were found when treatment purpose was compared; prevention/promotion (well populations), rehabilitation, or disease management. Three to 6 months post-baseline, a moderate positive effect of exercise interventions was found for overall QoL in rehabilitation patients, but no significant effect for well or disease management groups. However, physical and psychological QoL domains improved significantly relative to controls in well participants. Psychological QoL was significantly poorer relative to controls in the disease management group. This pattern of results persisted over 1 year. With some exceptions, better overall QoL was reported for light intensity exercise undertaken in group settings, with greater improvement in physical QoL following moderate intensity exercise. The implications for future health care practice and research are discussed. PMID- 19297066 TI - Social origins, early hardship and obesity: a strong association in women, but not in men? AB - This study investigates the relation between early life conditions and adult obesity in France, using a rich data set collected through the 2003 nationally representative Life History Survey. No salient factor emerged in men, while in women, after controlling for current socio-demographic characteristics, a relation was found between obesity and the following factors: father's occupation (OR=3.2 for women whose father was a clerical worker, versus those whose father was in a higher-level occupation); experience of economic hardship in childhood (OR=2.0), and; high parity (OR=2.1 for parities of more than 3 versus parity of 1). Neither early family history nor mother's working status surfaced as significant factors. Those findings highlight a definite gender pattern, with a strong association between early disadvantage and obesity in women, but not in men. Potential mechanisms are discussed, particularly the "habitus", the "thrifty phenotype" and the "feast-famine" hypotheses, and possible interactions with childbearing and motherhood. An integration of social and biological perspectives is needed to reach a better understanding of the processes involved, and to achieve progress in primary and secondary prevention. PMID- 19297068 TI - Leaning is common during in-hospital pediatric CPR, and decreased with automated corrective feedback. AB - BACKGROUND: Cardiopulmonary Resuscitation (CPR) guidelines recommend complete release between chest compressions (CC). No study has evaluated prevalence of leaning and the effect of real-time automated audiovisual feedback during in hospital pediatric CPR. OBJECTIVES: We hypothesize that leaning during in hospital pediatric CPR will be common, and that real-time automated feedback will be associated with reduced leaning prevalence and force. METHODS: A feedback capable monitor/defibrillator equipped with force transducer and accelerometer recorded CC leaning force and depth during in-hospital cardiac arrests (>/=8 years) at a children's hospital. Automated feedback was enabled at the resuscitation leader's discretion, and audiovisual prompts were given when leaning force exceeded 2.5 kg. Leaning force and depth CC with No Feedback (NoF) vs. with Feedback (F) were compared. RESULTS: 20 pediatric (mean age 14.7+/-3.8 years) pulse less arrests generated 37,396 evaluable CC. Median leaning force was 1.6 kg [0.9-2.7 kg] and leaning depth 2.9 (1.6-4.7)mm. Leaning force was greater with NoF (2.5 kg, [1.6-3.5 kg]; n=1921) vs. F (1.6 kg [0.9-2.6 kg]; n=35,164, p<0.001). Leaning>2.5 kg (adult feedback threshold) occurred in 50% (n=969) of CC with NoF and 27% (n=9367) CC with F (p<0.001). CC without leaning, defined as a leaning force of<0.5 kg, occurred in 2.2% (n=43) CC with NoF vs. 10.5% (n=3681) CC with F (p<0.001). CONCLUSIONS: Leaning (residual force>2.5 kg) was common during pediatric CPR. The prevalence and force of leaning were reduced with automated audiovisual feedback. Further study is necessary to determine the effect of the specific leaning threshold on CPR hemodynamics. PMID- 19297067 TI - Derangements in blood glucose following initial resuscitation from in-hospital cardiac arrest: a report from the national registry of cardiopulmonary resuscitation. AB - STUDY AIMS: Hyperglycemia is associated with poor outcomes in critically ill patients. We examined blood glucose values following in-hospital cardiac arrest (IHCA) to (1) characterize post-arrest glucose ranges, (2) develop outcomes-based thresholds of hyperglycemia and hypoglycemia, and (3) identify risk factors associated with post-arrest glucose derangements. METHODS: We retrospectively studied 17,800 adult IHCA events reported to the National Registry of Cardiopulmonary Resuscitation (NRCPR) from January 1, 2005 through February 1, 2007. RESULTS: Data were available from 3218 index events. Maximum blood glucose values were elevated in diabetics (median 226 mg/dL [IQR, 165-307 mg/dL], 12.5 mmol/L [IQR 9.2-17.0 mmol/L]) and non-diabetics (median 176 mg/dL [IQR, 135-239 mg/dL], 9.78 mmol/L [IQR 7.5-13.3 mmol/L]). Unadjusted survival to hospital discharge was higher in non-diabetics than diabetics (45.5% [95% CI, 43.3-47.6%] vs. 41.7% [95% CI, 38.9-44.5%], p=0.037). Non-diabetics displayed decreased adjusted survival odds for minimum glucose values outside the range of 71-170 mg/dL (3.9-9.4 mmol/L) and maximum values outside the range of 111-240 mg/dL (6.2 13.3 mmol/L). Diabetic survival odds decreased for minimum glucose greater than 240 mg/dL (13.3 mmol/L). In non-diabetics, arrest duration was identified as a significant factor associated with the development of hypo- and hyperglycemia. CONCLUSIONS: Hyperglycemia is common in diabetics and non-diabetics following IHCA. Survival odds in diabetics are relatively insensitive to blood glucose with decreased survival only associated with severe (>240 mg/dL, >13.3 mmol/dL) hyperglycemia. In non-diabetics, survival odds were sensitive to hypoglycemia (<70 mg/dL, <3.9 mmol/L). PMID- 19297069 TI - Positioning prior to endotracheal intubation on a television medical drama: perhaps life mimics art. PMID- 19297070 TI - Magnetic resonance imaging of three-dimensional cervical anatomy in the second and third trimester. AB - OBJECTIVE: Although a short cervix is known to be associated with preterm birth, the patterns of three-dimensional, anatomic changes leading to a short cervix are unknown. Our objective was to (1) construct three-dimensional anatomic models during normal pregnancy and (2) use the models to compare cervical anatomy in the second and third trimester. STUDY DESIGN: A cross-sectional study was performed in a population of patients referred to magnetic resonance imaging (MRI) for a fetal indication. Using magnetic resonance images for guidance, three-dimensional solid models of the following anatomic structures were constructed: amniotic cavity, uterine wall, cervical stroma, cervical mucosa and anterior vaginal wall. To compare cervical anatomy in the second and third trimester, models were matched according the size of the bony pelvis. RESULTS: Fourteen patients were imaged and divided into two groups according to gestational age: 20-24 weeks (n=7)) and 31-36 weeks (n=7). Compared to the second trimester, the third trimester was associated with significant descent of the amniotic sac (p=.02). Descent of the amniotic sac was associated with modified anatomy of the uterocervical junction. These three-dimensional changes were associated with a cervix that appeared shorter in the third trimester. CONCLUSION: We report a technique for constructing MRI-based, three-dimensional anatomic models during pregnancy. Compared to the second trimester, the third trimester is associated with three-dimensional changes in the cervix and lower uterine segment. PMID- 19297071 TI - Oxidative stress and epididymal sperm transport, motility and morphological defects. AB - OBJECTIVES: Radical oxidative species (ROS) have an important effect on sperm quality and quantity. Oxidative stress (OS) occurs when production of potentially destructive reactive oxygen species (ROS) exceeds the body's own natural antioxidant defenses, resulting in cellular damage. OS is a common pathology seen in approximately half of all infertile men. Increased ROS generation and reduced antioxidant capacity is negatively correlated with sperm concentration and motility in infertile men. For the first time, we used a more stable and reliable sensitive carbonyl protein (CP) detection method to determine ROS in seminal plasma than measuring ROS directly to clarify the effect of OS on spermatozoa in terms of protein dysfunction. This is the first report to measure CP in seminal plasma as an indicator of OS. Furthermore, for the first time we correlated the results of CP measurement with light microscopy in combination with ultrastructural analysis by electron microscopy. MATERIAL AND METHODS: 20 patients with idiopathic oligoasthenoteratozoospermia (iOAT) and 10 fertile controls were enrolled in this study. CP values were measured by enzyme-linked immuno sorbent assay (ELISA) to detect the level of OS. Transmission electron microscope (TEM) was used to detect axonemal anomalies. RESULTS: Compared to fertile controls, statistically highly significant higher degrees of abnormal sperm parameters (P<0.001) could be found in iOAT patients. CP values were highly significantly elevated in iOAT patients than in normal controls (P<0.001). A statistically highly significant difference in different axonemal anomalies were found between iOAT patients and normal controls (P<0.001). CP values have been found to be positively correlated with different axonemal anomalies (absence of axoneme (r(2)=0.841), missing of central singlet tubules (r(2)=0.702) and missing of outer doublet tubules (r(2)=0.869). A statistically negative correlation were found between different axonemal anomalies (absent axoneme (r(2)=-0.780), missing of central singlet tubules (r(2)=-0.611), and missing of outer doublet tubules (r(2)=-0.738) and forward progressive sperm motility. CONCLUSION: High levels of CP can be measured in iOAT patients, indicating that OS could underlie the aetipopathogenesis of the syndrome. OS negatively affects flagellar axonemal structure with subsequent impairment of forward progressive sperm motility. This can put an attention for antioxidants as a therapy for iOAT syndrome and further research to find how to decrease ROS production. PMID- 19297072 TI - Leiomyoma pseudocapsule after pre-surgical treatment with gonadotropin-releasing hormone agonists: relationship between clinical features and immunohistochemical changes. AB - OBJECTIVE: To evaluate if pre-operative GnRH-a modify uterine leiomyoma pseudocapsule and the possible clinical effects of these changes. STUDY DESIGN: The study was performed at the University Federico II of Naples on 33 premenopausal patients submitted to laparotomic myomectomy after treatment with triptorelin depot. 29 untreated patients formed the control group. The operating time, the intraoperative bleeding and the prompt identification of the cleavage plan between myoma and myometrium were evaluated. The pseudocapsule features and the immunoexpression of PCNA and CD34 in this area were studied. RESULTS: Treated patients showed lower blood loss and not clearly identifiable cleavage plan, but without any significant increase in the operating time. Treated lesions showed less evident border between myoma and myometrium and lower PCNA and CD34 pseudocapsule immunoexpression than untreated ones. CONCLUSION: We propose the changes of leiomyoma pseudocapsule as partial explanations of the reported clinical and surgical findings after pre-operative GnRH-a. PMID- 19297073 TI - Imaging of placental transport mechanisms: a review. AB - Functional analysis of material transfers requires precise statement of residence times in each tissue compartment. For the placenta, neither extractive biochemistry, isotope partitioning, nor mass-based quantitative assays provide adequate spatial resolution to allow the necessary precision. Dual-perfusion assays of material transfer in isolated placental cotyledons provide time-series data for two compartments, the maternal and fetal blood, but fail to distinguish the two cellular compartments (syncytiotrophoblast, fetal endothelium) which actively regulate rates of transfer in each direction for essentially every important molecule type. At present, no definitive technology exists for functional analysis of placental transfer functions. The challenge in developing such a technology lies in the exquisitely small and delicate structures involved, which are scaled at cellular and subcellular sizes (between 50 nm and 50 microm). The only available technologies attaining this high spatial resolution are imaging technologies, primarily light and electron microscopy. To achieve the high-quality images necessary, confocal laser scanning microscopy (CLSM) is required, to provide a uniform optical sectioning plane. In turn, this requires relatively high fluorescence intensities. Design of an adequate technology therefore bases on CLSM imaging fluorochrome-tagged tracers. The temporal resolution necessary to analyse placental material transfers is expected to be of the order of a few seconds, so that conventional wet-fixation protocols are too slow. For adequately rapid fixation, snap-freezing is required. As part of this review we report results obtained from an appropriately designed experimental protocol, analysed by CLSM and transmission electron microscopy (TEM). The images acquired were tested for uniformity of illumination and fluorescence emission strength. Relevant data was encoded in the green channel of the trichrome images obtained, and this was thresholded by application of strict quantitative criteria. The thresholding procedure is suitable for automation and produces reproducible, objectifiable results. Thresholded images were subjected to image calculation procedures designed to highlight image elements (pixels) containing (green) fluorescence associated with the tracer protein; all other sources of fluorescence were visualised in the final images only if no green fluorescence was detectable in that pixel. The resulting images were maps, showing the distribution of tracer molecules at a predefined time interval after perfusion of the tracer into the vital (term) cotyledon. Spatial resolution was routinely better than 1 microm and temporal resolution was approximately 5s. At timepoints up to 10 min after intravital application into the fetal vascular circulation, tracer was associated with capillaries in the villous structures, and no tracer was observed in the syncytiotrophoblast. Clear distinction was achieved between the four tissue compartments relevant to placental transfers, thus providing a novel technology capable of generating high-quality data concerning the regulation of transfers of any molecule that can bear a fluorescent tag. The potential applications of this methodology lie in analyses of factors influencing the rates of fetomaternal and maternofetal exchanges (for example, drugs), and of functional responses of the placental regulation to pathophysiological conditions such as hypoxia. PMID- 19297074 TI - The effects of the ovarian cycle and pregnancy on uterine vascular impedance and uterine artery mechanics. AB - OBJECTIVES: Uterine vascular resistance (UVR) is the ratio of systemic mean arterial pressure to mean uterine blood flow and is sensitive to changes in small arteries and arterioles. However, it provides little or no insight into changes in large, conduit arteries. Fluctuations in estrogen (E2) and progesterone (P4) levels during the ovarian cycle are thought to cause uterine resistance artery vasodilation; the effects on large arteries are unknown. Herein, our objective was to use the uterine vascular impedance, which is sensitive to changes in small and large arteries, to determine the effects of the ovarian cycle and pregnancy on the entire uterine vasculature. STUDY DESIGN: Uterine vascular perfusion pressure and flow rate were recorded simultaneously in anesthetized sheep in the nonpregnant (NP) luteal (NP-L, n=6) and follicular (NP-F, n=7) phases and in late pregnancy (CP, n=10). Impedance and metrics of impedance (input impedance Z(0), index of wave reflection R(W), characteristic impedance Z(C)) were calculated. E2 and P4 levels were measured from jugular vein blood samples. Finally, from pressure-diameter tests post-mortem, large uterine artery circumferential elastic modulus (E(Circ)) was measured. Significant differences were evaluated by two-way ANOVA or Student's t-test. RESULTS: As expected, E2:P4 was higher in the NP-F group compared to the NP-L group (p<0.05). Also as expected, UVR and Z(0) decreased in the follicular phase compared to the luteal (p<0.05), but R(W), Z(C), and E(Circ) were unaltered. Pregnancy not only substantially decreased UVR (and Z(0)) (p<0.00001) but also decreased Z(C) (p<0.001), R(W) (p<0.0001), E(Circ) (p<0.01), and pulse wave velocity (p<0.0001). CONCLUSIONS: The E2:P4 ratio mediates resistance artery vasodilatation in nonpregnant states, but has no effect on conduit artery size or stiffness. In contrast, pregnancy causes dramatic vasodilation and remodeling, including substantial reductions in conduit artery stiffness and increases in conduit artery size, which affect pulsatile uterine hemodynamics. PMID- 19297075 TI - The posterior intravaginal slingplasty operation: results of the Austrian registry. AB - OBJECTIVE: We set up a registry to assess complications and short-term results of the posterior intravaginal slingplasty operation. STUDY DESIGN: A total of 14 gynecology departments in Austria completed questionnaires addressing the patient's history, the operation itself and the postoperative course. In the follow-up we asked for information on tape exposure and functional and anatomical results. RESULTS: Fourteen centers entered a total of 577 patients operated between 2001 and 2006. 560 (97%) posterior slingplasty operations were done in conjunction with other procedures. Intraoperative complications were reported for 16 (2.8%) procedures. Postoperatively five hematomas required reoperation. 496 (86%) patients were available for follow-up after a median of 7 weeks (range, 1 156). 54 (9.4%) patients required reoperation. Vaginal tape exposure was seen in 50 (8.7%) women. Physicians assessed the functional and anatomical results as excellent or good in 83% and 88% of patients, respectively. CONCLUSION: Despite the limitations of a registry and the high rate of concomitant procedures, this study provides data on the complications and results of the posterior intravaginal slingplasty operation. PMID- 19297076 TI - Are infertile men less healthy than fertile men? Results of a prospective case control survey. AB - BACKGROUND: An association between either subfertility or infertility and an elevated risk of certain male cancers has been previously reported. Nothing is known about abnormalities in infertility and general health conditions. OBJECTIVE: To assess whether men with male factor infertility (MFI) are overall less healthy than fertile men, regardless of the reasons for infertility. DESIGN, SETTING, AND PARTICIPANTS: From September 2006 to September 2007, 344 consecutive European Caucasian men with MFI were enrolled in this prospective case-controlled study. Patients were compared with a control group of 293 consecutive age comparable fertile men. Infertile men were consecutively attending the outpatient male reproductive clinic at a tertiary academic center. Fertile controls were consecutively recruited by use of advertisements posted within our hospital. MEASUREMENTS: Comorbidities of patients and fertile men were objectively scored with the Charlson Comorbidity Index (CCI) according to the International Classification of Diseases modified ninth version (ICD-9-CM) codes. Multivariate linear regression models tested the association between predictors and CCI score, as a proxy of general health status. RESULTS: According to the CCI scores, infertile men had a significantly higher rate of comorbidities compared with the fertile controls (CCI: 0.33 [0.8] vs 0.14 [0.5]; p<0.001; 95% CI: 0.08-0.29). Linear regression analyses showed that although educational status did not have an impact on CCI (beta: 0.035; p=0.365), while CCI linearly increased with age (beta: 0.196; p<0.001) and body mass index (BMI; beta: 0.161; p<0.001). After adjusting for age, BMI, and educational status, a significantly lower CCI was calculated for fertile men and compared with MFI patients (beta: -0.199; p<0.001). CONCLUSIONS: These results show that MFI accounts for a higher CCI, which may be considered a reliable proxy of a lower general health status. PMID- 19297078 TI - Re: Paul L. Crispen, Michael L. Blute. Do percutaneous renal tumor biopsies at initial presentation affect treatment strategies? Eur Urol 2009;55:307-9. PMID- 19297079 TI - Pelvic lymph node dissection in prostate cancer. AB - CONTEXT: Pelvic lymph node dissection (PLND) is considered the most reliable procedure for the detection of lymph node metastases in prostate cancer (PCa); however, the therapeutic benefit of PLND in PCa management is currently under debate. OBJECTIVE: To systematically review the available literature concerning the role of PLND and its extent in PCa staging and outcome. All of the existing recommendations and staging tools determining the need for PLND were also assessed. Moreover, a systematic review was performed of the long-term outcome of node-positive patients stratified according to the extent of nodal invasion. EVIDENCE ACQUISITION: A Medline search was conducted to identify original and review articles as well as editorials addressing the significance of PLND in PCa. Keywords included prostate cancer, pelvic lymph node dissection, radical prostatectomy, imaging, and complications. Data from the selected studies focussing on the role of PLND in PCa staging and outcome were reviewed and discussed by all of the contributing authors. EVIDENCE SYNTHESIS: Despite recent advances in imaging techniques, PLND remains the most accurate staging procedure for the detection of lymph node invasion (LNI) in PCa. The rate of LNI increases with the extent of PLND. Extended PLND (ePLND; ie, removal of obturator, external iliac, hypogastric with or without presacral and common iliac nodes) significantly improves the detection of lymph node metastases compared with limited PLND (lPLND; ie, removal of obturator with or without external iliac nodes), which is associated with poor staging accuracy. Because not all patients with PCa are at the same risk of harbouring nodal metastases, several nomograms and tables have been developed and validated to identify candidates for PLND. These tools, however, are based mostly on findings derived from lPLND dissections performed in older patient series. According to these prediction models, a staging PLND might be omitted in low-risk PCa patients because of the low rate of lymph node metastases found, even after extended dissections (<8%). The outcome for patients with positive nodes is not necessarily poor. Indeed, patients with low-volume nodal metastases experience excellent survival rates, regardless of adjuvant treatment. But despite few retrospective studies reporting an association between PLND and PCa progression and survival, the exact impact of PLND on patient outcomes has not yet been clearly proven because of the lack of prospective randomised trials. CONCLUSIONS: On the basis of current data, we suggest that if a PLND is indicated, then it should be extended. Conversely, in view of the low rate of LNI among patients with low-risk PCa, a staging ePLND might be spared in this patient category. Whether this approach is also safe from oncologic perspectives is still unknown. Patients with low-volume nodal metastases have a good long-term prognosis; to what extent this prognosis is the result of a positive impact of PLND on PCa outcomes is still to be determined. PMID- 19297080 TI - Editorial comment on: Are infertile men less healthy than fertile men? Results of a prospective case-control survey. PMID- 19297081 TI - Validation of the Chinese non-motor symptoms scale for Parkinson's disease: results from a Chinese pilot study. AB - OBJECTIVES: To evaluate a Chinese version of the Non-Motor Symptoms Scale (NMSS) in Parkinson's disease (PD) as an instrument for measuring non-motor symptoms (NMSs) in Chinese patients with Parkinson's disease. METHODS: We conducted a psychometric analysis of the Chinese version of NMSS using a cross-sectional study of 126 patients with PD. The battery also included the Pittsburgh Sleep Quality Index (PSQI), the Epworth Sleepiness Scale (ESS), the Mini-Mental State Examination (MMSE), the Geriatric Depression Scale (GDS), and the Hamilton Anxiety Scale (HAMA), and was conducted by neurologists. RESULTS: There were significant correlations between the NMSS and PSQI scores (rS=0.63, P<0.001), as well as the NMSS and ESS scores (rS=0.38, P<0.001). Furthermore, significant positive correlations between NMSS and GDS, NMSS and HAMA, and NMSS and disease duration were also observed. Importantly, the sleep/fatigue index of the NMSS significantly correlated with the PSQI and ESS findings, the mood/cognition index of the NMSS significantly correlated with the GDS and HAMA findings, and the attention/memory index of the NMSS significantly correlated with the MMSE findings. CONCLUSION: The Chinese version of the NMSS can be considered a comprehensive, useful measure for NMS evaluation in Chinese PD patients. PMID- 19297082 TI - Intramedullary cavernous malformations: clinical features and surgical technique via hemilaminectomy. AB - OBJECTIVE: The purpose of this study was to define the clinical features and the surgical technique of unilateral hemilaminectomy for treating intramedullary cavernous malformations. MATERIALS AND METHODS: Retrospective chart was performed in 16 patients with histologically diagnosed intramedullary cavernous malformations. All patients were treated with unilateral hemilaminectomy and microsurgical resection of the malformations. The pre- and postoperative neurological state was evaluated using Frankel scale. RESULTS: There were nine females and seven males (mean age 38 years) harbouring symptomatic intramedullary cavernous malformations. The annual retrospective haemorrhage rate was 3.1% per patient/year. All cavernous malformations were completely resected. Twelve of 16 patients experienced the improvement of the neurological state and in four patients, clinical features remained unchanged during the follow-up period. Static and dynamic plain radiograph film showed none of them had spinal deformity or spinal instability. CONCLUSION: According to the defined bleeding risk, symptomatic and MRI-morphologically growing intramedullary cavernous malformations should be totally surgically removed, to avoid the recurrence and rebleeding of the residue. A least traumatic myelotomy, as well as a meticulous microsurgical technique and the intraoperative somatosensory evoked potentials monitoring, together with selection of a minimally invasive microsurgical approach (hemilaminectomy), leads to a favourable outcome and prevents additional morbidity. PMID- 19297083 TI - Fibrinolytic therapy versus craniotomy for anticoagulant-associated intracerebral hemorrhage. AB - OBJECT: Anticoagulant-associated intracerebral hemorrhages (AAICH) have a high morbidity and mortality, necessitating urgent treatment. We examined outcomes after conventional craniotomy and stereotactic fibrinolytic therapy in a series of patients with anticoagulant-associated hemorrhages. METHODS: Among 129 consecutive surgically treated patients with supratentorial intracerebral hemorrhage, 27 patients with AAICH were identified (mean age 62; range 36-79). Thirteen patients underwent craniotomy for surgical hematoma evacuation, and 14 patients hematoma puncture and catheter placement for clot lysis. The groups had comparable major prognostic factors such as hematoma volume, age, and Glasgow coma scale (GCS) score at admission. RESULTS: Nine patients died despite treatment (mortality=33%). Mortality in the craniotomy group was comparable to that of the lysis group (46% versus 21%; p=0.13). Good outcomes (Glasgow outcome score of 4 or 5) were seen in 3 craniotomy patients (23%) and 2 fibrinolysis patients (14%). Half the patients survived with major neurological deficits (GOS 2 or 3) (n=13; 48%). One rebleed was observed two days after uneventful craniotomy and hematoma removal, while no patient who underwent fibrinolysis had rebleeding. CONCLUSIONS: Approximately one-fifth of patients with AAICH managed surgically may have good outcomes. Mortality and favourable outcome rates are comparable between craniotomy and fibrinolytic therapy. Fibrinolytic therapy appears to be a reasonable less invasive alternative treatment modality for intracerebral hemorrhage in the anticoagulated patient. PMID- 19297084 TI - Dynamics of PM2.5 concentrations in Kathmandu Valley, Nepal. AB - This study analyzed daily patterns and dynamics of PM(2.5) concentrations in the Kathmandu Valley during three winters. The PM(2.5) data showed a daily repetitive cycle which represents influence of local air flow and dispersion and accumulation of air pollutants in the valley. Two concentration peaks were observed in the morning and in the evening periods, and they fell down during the daytime and the nighttime periods. This indicates local emission sources as major contributors in the valley. The more pronounced morning peak compared to the evening peak showed that the upslope wind in the morning helped to move the polluted inversion layer downward, subsequently adding to freshly emitted pollutants and causing a sharp pollutant concentration rise in the morning. Katabatic wind and rise of temperature in the basin during the day helped the pollutant upflow and dilution, resulting in a sharp PM(2.5) concentration decline. Through the afternoon, the decrease in air temperature followed by decrease in wind speed caused to lower PM(2.5) peaks in the evening. Also, higher morning peaks of PM(2.5) concentrations compared to the evening indicated pollution from the previous day is added to the fresh emission. The valley had increased PM(2.5) from the beginning of October which continued till the first week of February. The increase in PM(2.5) peak fit the logistic equation y=[k/(1+exp (p-qx)]+asin(bx) where k, p, q, a, and b are constants. PMID- 19297085 TI - Advanced oxidation and mineralization of simazine using Fenton's reagent. AB - Removal of simazine from aqueous solution by Fenton's reagent oxidation was investigated. Box-Behnken statistical experiment design and the response surface methods were used to investigate the effects of simazine, H(2)O(2) and Fe (II) concentrations on simazine degradation and mineralization. Total organic carbon (TOC) and simazine removals were investigated at different reagent doses to determine the experimental conditions yielding the highest removal of simazine and TOC. Fe (II) concentration had more profound effect than H(2)O(2) for simazine removal while all parameters affected mineralization (TOC removal). Complete disappearance of simazine was achieved within 6 min reaction period. However, only 32% of simazine was mineralized after 15 min indicating formation of some intermediate products. The optimal H(2)O(2)/Fe (II)/simazine ratio resulting in the maximum pesticide (100%) and TOC removal (32%) was found to be 55/15/3 (mg L(-1)). The initial rate of simazine degradation was found to be first-order with respect to the initial simazine concentration. PMID- 19297086 TI - Removal of Cu(II) from aqueous solution by agricultural by-product: peanut hull. AB - Peanut hull, an agricultural by-product abundant in China, was used as adsorbent for the removal of Cu(II) from aqueous solutions. The extent of adsorption was investigated as a function of pH, contact time, adsorbate concentration and reaction temperature. The Cu(II) removal was pH-dependent, reaching a maximum at pH 5.5. The biosorption process followed pseudo-second-order kinetics and equilibrium was attained at 2h. The rate constant increased with the increase of temperature indicates endothermic nature of biosorption. The activation energy (E(a)) of Cu(II) biosorption was determined at 17.02kJ/mol according to Arrhenius equation which shows that biosorption may be an activated chemical biosorption. Other activation parameters such as DeltaH(#), DeltaS(#), and DeltaG(#) were also determined from Eyring equation. The equilibrium data were analyzed using the Langmuir, Freundlich, and Dubinin-Radushkevich (D-R) isotherm models depending on temperature. The equilibrium biosorption capacity of Cu(II) determined from the Langmuir equation was 21.25mg/g at 30 degrees C. The mean free energy E (kJ/mol) got from the D-R isotherm also indicated a chemical ion-exchange mechanism. The thermodynamic parameters such as changes in Gibbs free energy (DeltaG(0)), enthalpy (DeltaH(0)) and entropy (DeltaS(0)) were used to predict the nature of biosorption process. The negative DeltaG(0) values at various temperatures confirm the biosorption processes are spontaneous. PMID- 19297087 TI - Effect of natural organic matter on arsenic mobilization from mine tailings. AB - This research study was to elucidate the mechanisms of arsenic mobilization from mine tailings in the presence of natural organic matter (NOM). Humic acid (HA) was chosen as a model for NOM. The introduction of the HA at a low mass ratio (below 2mg added HA/g mine tailings) inhibited arsenic mobilization under acidic conditions. Arsenic mobilization increased with increasing mass ratio. When pH was above 7, the addition of HA enhanced arsenic mobilization significantly. A mobilization isotherm was developed to predict arsenic mobilization from the mine tailings in the presence of HA. It was indicated that HA sorption to the mine tailings was essential for arsenic mobilization. HA might enhance arsenic mobility through formation of aqueous complexes, competition for adsorption and electrostatic interactions. Capillary electrophoresis analyses indicated that arsenic redox reactions might not have a significant effect on arsenic mobilization in this study. The mobilization of co-existing metals could enhance arsenic mobilization by helping incorporating it into soluble complexes in the presence of HA. PMID- 19297088 TI - Removal of Cu2+ and Zn2+ from model wastewaters by spontaneous reduction coagulation process in flow conditions. AB - The treatment of model wastewaters containing heavy metals by spontaneous reduction-coagulation process using micro-alloyed aluminium composite (MAlC) in a laboratory semi-flow system (SFS) has been investigated. Several working parameters, such as treatment time, pH, initial metal concentration and flow rate were studied in an attempt to achieve a higher removal capacity. The residual concentrations of metals were at admissible levels after only 20 min of treatment. Removal rate of Zn(2+) was greater at neutral pH than at acid or basic, while Cu(2+) removal was less affected by pH. Removal constants of both metals decreased as initial metal concentration increased from 20 to 200 mg L( 1). In mixed wastewaters the presence of copper caused increase of Zn(2+) removal efficacy, however, the copper removal was not affected by zinc. The removal mechanisms depend on metals nature and pH: copper was mainly removed by reduction, while zinc by precipitation as hydroxide. The kinetic of process was mass-transport limited, thus increasing of flow rate accelerated removal of metals. The method was found to be highly efficient and faster compared to conventional treatments. PMID- 19297089 TI - Treatment of coking wastewater by using manganese and magnesium ores. AB - This study investigated a wastewater treatment technique based on natural minerals. A two-step process using manganese (Mn) and magnesium (Mg) containing ores were tested to remove typical contaminants from coking wastewater. Under acidic conditions, a reactor packed with Mn ore demonstrated strong oxidizing capability and destroyed volatile phenols, chemical oxygen demand (COD)(,) and sulfide from the coking wastewater. The effluent was further treated by using Mg ore to remove ammonium-nitrogen and phosphate in the form of magnesium ammonium phosphate (struvite) precipitates. When pH of the wastewater was adjusted to 1.2, the removal efficiencies for COD, volatile phenol and sulfide reached 70%, 99% and 100%, respectively. During the second step of precipitation, up to 94% of ammonium was removed from the aqueous phase, and precipitated in the form of struvite with phosphorus. The struvite crystals showed a needle-like structure. X ray diffraction and transmission electron microscopy were used to characterize the crystallized products. PMID- 19297090 TI - Effect of sample pretreatment on speciation of copper and zinc in MSW. AB - Copper and zinc were determined in MSW samples collected from Tianziling landfill site in Hangzhou, Zhejiang, east China by modified BCR sequential extractions. Three pretreatment methods, including fresh, air drying, and oven drying, were studied. It showed that the main cause of fraction transfer after drying could be ascribed to the variation of sample status, including the evaporation of ammonia nitrogen and volatile fatty acid (VFA), the shift of sample pH, and the oxidation of sulfur, when contacted with atmospheric oxygen or exposed in high temperature during drying processes. Effect of sample pretreatment methods on speciation of copper and zinc in MSW concluded that oven drying should not be used because the changes are more numerous and generally of greater amplitude, air drying was a 'neutral' treatment relatively, while fresh sample might be the best choice if possible. To validate the effect and determine the source of pretreatment methods clearly, certified reference materials of MSW should be confirmed in the future research. PMID- 19297091 TI - Feasibility of nutrient recovery from industrial sludge by vermicomposting technology. AB - Transformation of industrial sludges into vermicompost is of double interest: on the one hand, a waste is converted into value added product, and, on the other, it controls a pollutant that is a consequence of increasing industrialization. This paper reports the feasibility of utilization of vermicomposting technology for nutrient recovery from industrial sludge in laboratory scale experiment employing Eisenia fetida earthworm. A total of nine vermireactors, having different percentage of wastewater treatment plant sludge of a food industry and cow dung, were established and monitored for fertilizer quality of vermicompost and growth and fecundity of the earthworms for 3 months. The earthworms were unable to survive in 100% FIS. There was a decrease in pH, organic carbon content, organic matter, C:N ratio, and increase in ash content, EC, nitrogen, potassium and phosphorus content in all the vermireactors. Total Kjeldhal nitrogen (TKN) content increased in the range of 12.2-28.7 g kg(-1) in different vermireactors after vermicomposting. C:N ratio was 1.59-5.24 folds lesser in final vermicomposts than initial raw substrate. The heavy metals' content in final vermicomposts was higher than initial feed mixtures. Maximum worm biomass was observed in control, i.e., 100% CD (836 mg earthworm(-1)) and the lowest in 30% CD+70% FIS feed mixture (280 mg earthworm(-1)). Cocoon production was started during 6-7th week in all feed mixture except in feed mixture no. 9. After 12 weeks maximum cocoons (57) were counted in 100% CD and minimum (2) in 30% CD+70% FIS feed. The results indicated that food industry sludge could be converted into good quality manure by vermicomposting if mixed up to 30% with cow dung. PMID- 19297092 TI - Removal of phosphate from polluted water by lanthanum doped vesuvianite. AB - The adsorption capacities of vesuvianite and lanthanum doped vesuvianite were studied. The effects of different mass ratios of La/vesuvianite at different contact times, pHs, and temperatures on adsorption capacity were also studied. It was found that lanthanum doped vesuvianite exhibited higher adsorption capacity than undoped one due to the reaction of bounded lanthanum with phosphate. The adsorption capacity of lanthanum doped vesuvianite for phosphate removal increased with the increase of La/vesuvianite mass ratio. The Freundlich and Langmuir models were used to simulate the sorption equilibrium, and the results indicate that the Langmuir model had a better correlation with the experimental data than the Freundlich model did. When the initial phosphate concentration was 1mgP/L, the adsorptive capacity rate would be 1.32 mg P/g lanthanum doped vesuvianite (La/vesuvianite mass ratio >or=0.14) at pH between 6 and 9 after 40h. The concentrations of residual lanthanum ions in solution at different conditions were measured. Lanthanum doped vesuvianite was also used for the removal of phosphate in a polluted river water and it could be easily recycled once without losing its activity to a greater extent. PMID- 19297093 TI - Biodegradation of chlorpyrifos and its hydrolysis product 3,5,6-trichloro-2 pyridinol by Bacillus pumilus strain C2A1. AB - A bacterial strain C2A1 isolated from soil was found highly effective in degrading chlorpyrifos and its first hydrolysis metabolite 3,5,6-trichloro-2 pyridinol (TCP). On the basis of morphology, physiological characteristics, biochemical tests and 16S rRNA sequence analysis, strain C2A1 was identified as Bacillus pumilus. Role of strain C2A1 in the degradation of chlorpyrifos was examined under different culture conditions like pH, inoculum density, presence of added carbon/nutrient sources and pesticide concentration. Chlorpyrifos was utilized by strain C2A1 as the sole source of carbon and energy as well as it was co-metabolized in the presence of glucose, yeast extract and nutrient broth. Maximum pesticide degradation was observed at high pH (8.5) and high inoculum density when chlorpyrifos was used as the sole source and energy. In the presence of other nutrients, chlorpyrifos degradation was enhanced probably due to high growth on easily metabolizable compounds which in turn increased degradation. The strain C2A1 showed 90% degradation of TCP (300 mg L(-1)) within 8 days of incubation. PMID- 19297094 TI - Size effects of nanocrystalline TiO2 on As(V) and As(III) adsorption and As(III) photooxidation. AB - The physicochemical properties of TiO(2) particles in the diameter range between 6.6 and 30.1 nm and the effect of the crystalline size on arsenic adsorption and photocatalytical oxidation were investigated. TiO(2) nanoparticles of different sizes were single-phase anatase. The adsorption capacity of the TiO(2) for As(III) and As(V) increased linearly with the N(2) Brunauer-Emmett-Teller surface area (S(BET)) of the particles. There was not much difference in the rate of As(III) photooxidation when the diameter of the TiO(2) nanoparticles was between 6.6 and 14.8 nm. However, the As(III) photooxidation rate clearly decreased when the particle size increased to 30.1 nm. Arsenite photooxidation data could be fitted with a first-order kinetics equation. PMID- 19297095 TI - Radiation-induced grafting of glycidyl methacrylate onto cotton fabric waste and its modification for anchoring hazardous wastes from their solutions. AB - Ion exchange adsorbents based on cellulosic fabric wastes carrying sulfonic acid and amine functional groups were synthesized by radiation-induced graft polymerization of glycidyl methacrylate (GMA) with subsequent chemical modification of the epoxy groups of poly-GMA graft chains with sodium sulfite/H(2)SO(4) and triethylamine, respectively. The conversion of epoxy groups into the functional groups was investigated. Factors affecting on grafting process such as radiation dose, monomer concentration and solvent were studied. The synthesized adsorbent and its applications in the removal of different types of hazardous pollutants e.g. acidic dye, cobalt, dichromate and phenols from aqueous solution were also studied. PMID- 19297096 TI - Potential formation of PCDD/Fs and related bromine-substituted compounds from heating processes for ashes. AB - Thermal experiments were conducted using real boiler ash and fly ash samples from three types of municipal or industrial solid waste incineration plants to understand the formation reactions of polychlorinated dibenzo-p-dioxin and furans (PCDD/Fs) and related bromine compounds that were chlorinated-brominated dibenzodioxins and furans (PXDD/Fs) and polybrominated dibenzo-p-dioxin and furans (PBDD/Fs). The results obtained were as follows: The formation of PCDD/Fs was clearly shown, and fly ash containing abundant carbon matter had a significant potential for de novo synthesis. The homologous distribution change apparently showed that the formation of PXDD/Fs occurred from the substitution of a bromine atom with a chlorine atom in the PCDD/F molecules. This suggests that PXDD/Fs are usually formed with PCDD/Fs on the ash. PBDD/Fs might be formed from any reaction mechanism different from that of PXDD/Fs. The existence of carbonaceous matters always does not mean the potential formation of PCDD/Fs. However, any addition of catalytic copper may influence the nature of ash to increase the formation potential. The findings suggest that there are many instances that result in the unintended production of trace hazardous pollutants in the incineration process and show that careful and sophisticated control is required to prevent the formation of pollutants. PMID- 19297097 TI - Characterization of gas concrete materials used in buildings of Turkey. AB - The activity concentration of (226)Ra, (232)Th and (40)K in gas concrete samples collected from different suppliers and some provinces in Turkey were measured using gamma-ray spectrometry. Knowledge of radioactivity in gas concrete used in building materials enables one to assess any possible radiological risks to human health. The mean activity concentrations observed in the gas concrete samples were 82.0, 28.2 and 383.9 Bq kg(-1) for (226)Ra, (232)Th and (40)K, respectively. The radium equivalent activity, external and internal hazard indices as well as terrestrial absorbed dose and annual effective dose rate was calculated. The results indicate that the radium equivalent activity values of gas concrete samples are lower than the limit of 370 Bq kg(-1), equivalent to a gamma-dose of 1.5 mSv y(-1). Moreover, mass attenuation coefficients were measured in some gas concrete samples. It was found that the mass attenuation coefficients decreased with increasing photon energies. Also, chemical compositions and structural analysis (XRD and SEM) of the gas concrete samples were investigated. PMID- 19297098 TI - Cation exchange removal of Pb from aqueous solution by sorption onto NiO. AB - This paper reports NiO as a novel and an efficient adsorbent for the removal of Pb from aqueous solutions. In the present investigation, Pb adsorption experiments on NiO were conducted on aqueous solution at different initial Pb concentration, pH of the solution and adsorption temperature. The mechanism of adsorption was observed to be an ion exchange between the surface proton and Pb in aqueous solution. Experimental data were best described by the Langmuir isotherms. The surface structure of the NiO before and after adsorption of Pb was analyzed by using FTIR spectroscopy, scanning electron microscopy (SEM) coupled with energy dispersive X-ray analyses (EDX). PMID- 19297099 TI - A molecular survey of Piroplasmida and Hepatozoon isolated from domestic and wild animals in Burgos (northern Spain). AB - This study reports a molecular survey of Hepatozoon species and of the order Piroplasmida in the Province of Burgos, northern Spain. The diagnostic techniques employed included PCR and the sequencing of the 18S rRNA gene. Eighty-nine blood samples from domestic animals plus 138 blood/coagulated blood samples from wild mammals were examined. Theilerid protozoa were found at relatively high frequencies in bovines (14.6%) and horses (36%). Theileria buffeli, T. sergenti and T. annulata were diagnosed in cows. T. equi was common in horses and T. annae was found in a donkey for the first time. A new piroplasmid was found in the European badger (20%). This appears to be distantly related to both T. annae and a piroplasmid isolated from Lontra canadensis. A moderate prevalence (14%) for T. annae was recorded in red foxes. A species of hepatozoon was found in one bank vole (17%), while 28% of the red foxes examined were found to be infected with H. canis. Twenty-five wild house mice were studied and found not to be carriers of piroplasmids or Hepatozoon species. Wild boars, roe deer, hares, Apodemus sp. and moles were also negative for haematozoan infection. The present study indicates that piroplasmid protozoa are present at a low to moderate frequency in some domestic herbivores in the Burgos area. They also infect certain wild mammalian species, which may act as zoonotic carriers. PMID- 19297100 TI - Validation of the EQ-5D questionnaire in burn injured adults. AB - BACKGROUND: Health-related quality of life (HRQoL) is an important aspect of adaptation after burn. The EQ-5D is a standardized generic instrument for assessing HRQoL. Its psychometric properties in a group of burn injured individuals are, however, not known. METHODS: Seventy-eight consecutive patients admitted to a burn unit were included in a prospective longitudinal study. The participants completed the EQ-5D during acute care, and at 3, 6, and 12 months after the burn. At 6 and 12 months after the burn they also completed the Short Form 36 Health Survey (SF-36) and the Burn Specific Health Scale-Brief (BSHS-B). RESULTS: High feasibility of the EQ-5D was demonstrated through a high response rate and a low proportion of missing or invalid answers. The floor and ceiling effects were small. Construct validity was demonstrated through good differentiation between health states and good discrimination of health states over time. The EQ-5D was associated with burn severity and discriminated between clinical subgroups in an expected manner. Criterion validity was demonstrated through significant correlations between the EQ-5D and subscales of the SF-36 and the BSHS-B. CONCLUSIONS: The EQ-5D has good psychometric properties, it is short and easy to administer and thus useful in assessment of HRQoL after burn. PMID- 19297101 TI - Burns surgery handover study: trainees' assessment of current practice in the British Isles. AB - INTRODUCTION: Effective handover of clinical information between working shifts is essential for patient safety. The aim of this study was to identify current practice and trainees' assessment of handover in the burns units of the British Isles. METHODS: A telephone questionnaire was conducted to trainee burns surgeons (at junior and senior grades) currently working at all 30 burns surgery units in the British Isles. Information regarding timing, location, duration, participation and quality of handover was collated anonymously. Trainees commented on satisfaction with current practice and its perceived safety. RESULTS: A 100% response from all 30 units was obtained. 23/30 units (76.7%) had junior to junior trainee handovers. 17/30 (56.7%) had senior to senior trainee handovers. 19/30 units (63.3%) reported that handover took place with more than one grade of doctor present (range 1-4 grades). 3/30 (10%) reported that handover was bleep-free. 3/30 (10%) had received formal training on good burns handover. 5/30 (16.7%) were working in a unit that operated a "burns surgeon of the week" pattern of emergency cover. Mean satisfaction level was 3.8 out of 5. Those working in "surgeon of the week" teams had significantly higher scores, 4.4 versus 3.68 (p=0.037). Other healthcare professionals were present at only 4/30 (13.3%) handovers. Overall 26/30 (86.7%) of trainees judged their current handover practice "safe" (100% in "surgeon of week" group and 84% in the remaining group, p=0.289). CONCLUSIONS: Effective handover remains a keystone in safe and effective communication between doctors. The study highlights areas for improvement in handover practice, including greater involvement of an integrated multidisciplinary team. Those working under the "surgeon of the week" pattern are more satisfied. PMID- 19297102 TI - A randomized double-blind trial of the effects of hormone therapy on delayed verbal recall in older women. AB - We examined whether estradiol and norethindrone hormone therapy (HT) prevented decline in delayed verbal recall in older women with normal to mildly impaired memory functioning. This was a 2-year, randomized, double-blind, placebo controlled trial of 142 women aged 61-87, randomly assigned to receive 1 mg 17 beta estradiol daily and 0.35 mg norethindrone 3 days/week or daily placebo for 2 years. The primary outcome was short-delay verbal recall of the California Verbal Learning Test (CVLT). To look for differences in response to HT by baseline short delay recall, we examined the primary outcome in participants grouped according to whether their baseline scores were below average for the age group or greater than or equal to this score and according to whether they met criteria for Mild Cognitive Impairment (MCI) or not. 133 women completed 1 year of the trial and 128 completed 2 years. Prespecified covariates in all repeated measures analyses of covariance (RANCOVA) included age, education, APOE epsilon4, and prior HT use. RANCOVA showed no overall significant treatment effects at year 1 or year 2. After testing for an interaction, which was significant (p=0.02), we found that women in the HT group who scored at or above the average showed significantly less decline than the placebo group in short-delay verbal recall after 1 year, p=0.007 and 2 years, p=0.01. No treatment effects were found in women below the average in either year. When grouped according to whether the participant met criteria for MCI, the interaction between treatment group and MCI subgroup was not significant. These results suggest that benefits of estrogen exposure may be limited to those with average to above average scores on the delayed verbal recall. HT dose and formulation may have contributed to these beneficial outcomes. Replication is warranted before recommendations can be made in the clinical setting. PMID- 19297103 TI - Effects of sex hormonal levels and phases of the menstrual cycle in the processing of emotional faces. AB - Several neuropsychiatry disorders have shown a sexual dimorphism in their incidence, symptom profile and therapeutic response. A better understanding of the impact of sex hormones in emotional processing sexual dimorphism could bring light to this important clinical finding. Some studies have provided evidence of sex differences in the identification of emotional faces, however, results are inconsistent and such inconsistency could be related to the lack of experimental control of the sex hormone status of participants. More recently, a few studies evaluated the modulation of facial emotion recognition by the phase of the menstrual cycle and sex hormones, however, none of them directly compared these results with a group of men. We evaluated the accuracy of facial emotion recognition in 40 healthy volunteers. Eleven women were assigned to early follicular group, nine women to the ovulatory group and 10 women to luteal group, depending on the phase of menstrual cycle, and a group of 10 men were also evaluated. Estrogen, progesterone and testosterone levels were assessed. The performance of the groups in the identification of emotional faces varied depending on the emotion. Early follicular group were more accurate to perceive angry faces than all other groups. Sadness was more accurately recognized by early follicular group than by luteal group and regarding the recognition of fearful faces a trend to a better performance and a significantly higher accuracy was observed, respectively, in the early follicular group and in the ovulatory group, in comparison to men. In women, estrogen negatively correlated to the accuracy in perception of angry male faces. Our results indicate sex hormones to be implicated in a sexual dimorphism in facial emotion recognition, and highlight the importance of estrogen specifically in the recognition of negative emotions such as sadness, anger and fear. PMID- 19297104 TI - Reflections on a scientific paper of 1926 by the medical 'Inkling' Robert Emlyn 'Humphrey' Havard (1901-1985). AB - Robert Emlyn Havard (1901-1985; general practitioner and sometimes medical scientist) was the only non-literary member of the Inklings - a 1930 s and 1940s Oxford University club which included Lewis and Tolkien. Despite spending most of his time in family medicine, Havard was a productive medical scientist. While still a student at Cambridge University, Havard co-authored an influential study published in the Journal of Physiology of 1926 entitled 'The influence of exercise on the inorganic phosphates in the blood and urine'. The style and structure of this paper provides a charming window into the elite medical science of the 1920s. PMID- 19297105 TI - Adjuvant therapy for gallbladder carcinoma: the Mayo Clinic Experience. AB - PURPOSE: To analyze the effect of adjuvant chemoradiotherapy on gallbladder carcinoma. METHODS AND MATERIALS: We retrospectively reviewed the records from consecutive patients who underwent R0 resection of gallbladder carcinoma between January 1, 1985, and December 31, 2004. Patients had either Stage I (T1-T2N0M0) or Stage II (T3N0M0 or T1-T3N1M0) disease. Patients undergoing adjuvant therapy received 5-fluorouracil chemotherapy concurrently with radiotherapy (median dosage, 50.4 Gy in 28 fractions). Adverse prognostic factors and the effect of adjuvant treatment on overall survival (OS) were evaluated. RESULTS: A total of 73 patients were included in the analysis; of these, 25 received adjuvant chemoradiotherapy. On univariate analysis, no adverse prognostic factors for OS reached statistical significance, but trends were noted for Stage N1 vs. N0 (p = .06), Nx vs. N0 (p = .09), Stage T3 vs. T1-T2 (p = .06), and histologic findings other than adenocarcinoma (p = .13). The median OS for patients receiving adjuvant chemoradiotherapy vs. surgery alone was 4.8 years and 4.2 years, respectively (log-rank test, p = .56). However, a significantly greater percentage of patients receiving adjuvant chemoradiotherapy had Stage II disease (p <.001). In the multivariate Cox model, increasing T and N category and histologic findings other than adenocarcinoma were significant predictors of decreased OS. Additionally, adjuvant chemoradiotherapy was a significant predictor of improved OS after adjusting for these prognostic factors (hazard ratio for death, 0.3; 95% confidence interval, 0.13-0.69; p = .004). CONCLUSION: After adjusting for the stage parameters and histologic findings, our data suggest that adjuvant chemoradiotherapy might improve OS for patients with gallbladder cancer. PMID- 19297106 TI - A validated hybrid quadrupole linear ion-trap LC-MS method for the analysis of morphine and morphine glucuronides applied to opiate deaths. AB - A hybrid quadrupole linear ion-trap mass spectrometer using an electrospray ionisation ion source coupled to a HPLC system has been used to develop a method which can accurately measure morphine, morphine-3-glucuronide (M3G) and morphine 6-glucuronide (M6G) in plasma, whole blood and post-mortem blood following solid phase extraction. The method can also qualitatively detect various other opioids and related compounds including: codeine, dihydrocodeine (and metabolites), noscapine, papaverine and 6-acetylmorphine (6-AM). The method has been favourably compared to an existing laboratory method using a now discontinued radio immunoassay technique. The advantage of measuring the glucuronides directly rather than following deconjugation by beta-glucuronidase has also been shown. Detection and quantification of compounds was achieved using multiple reaction monitoring (MRM) incorporating the use of deuterated morphine and M3G as internal standards. Precision and accuracy was determined to be less than 10% at both high and low levels for all analytes and the calibration curve was deemed linear over an acceptable range. Recovery in blood was greater than 90% and ion suppression/enhancement was shown to be less than 15%. This method was applied to over 130 post-mortem cases involving the use of heroin, prescribed morphine and codeine. The range of concentrations of morphine, M3G and M6G was large (particularly in heroin and prescribed morphine cases), reflecting the many different factors involved with therapeutic use or fatal opiate poisonings, including tolerance associated with regular use, variable dose regimens and co administration of other drugs. Detection of other constituents of the opium poppy such as noscapine and papaverine and metabolites of diacetylmorphine in the blood (6-AM) was useful in determining the source of the morphine (i.e. illicit heroin) and the rapidity of death after administration. PMID- 19297107 TI - Discolouration of the brain as the only remarkable autopsy finding in hydrogen sulphide poisoning. PMID- 19297108 TI - Comparison of surgical outcomes of endonasal dacryocystorhinostomy with or without mucosal flaps. AB - OBJECTIVE: The goals of this study were to evaluate the long-term results of endoscopic endonasal dacryocystorhinostomy (DCR) with or without a posterior mucosal flap and to compare the surgical success rates of that procedure in patients with a nasolacrimal duct obstruction. PATIENTS AND METHODS: We retrospectively investigated the results of 78 endoscopic endonasal DCRs performed at the Baskent University, Department of Otolaryngology between December 2000 and March 2007 on 74 patients with a lacrimal obstruction. The patients were divided into two groups. During surgery, the posterior mucosal flap was preserved in 27 patients (group A) and removed in 47 patients (group B). All patients underwent intubation with a silicone tube at the conclusion of surgery. The silicone tube was removed within 6 months after surgery. The mean follow-up period was 36 months (range, 2-78 months). The results obtained were then compared. RESULTS: Granulation tissue and synechia developed between the lateral nasal wall and the middle turbinate in one patient in group A (revision surgery was not required). In group B, granulation tissue at the rhinostomy opening was found in seven patients, and in four of those subjects, the granulation tissue obstructed the neo-ostium. These four patients underwent a second operation. In group B, synechia was noted between the middle turbinate and the lateral nasal wall in two of 47 patients. In group A, the procedure was successful for all patients except one in whom granulation tissue developed, and in group B, the surgical success rate was 88.3%. CONCLUSION: In endoscopic endonasal DCR, the closure of bare bone with a posteriorly based nasal mucosal flap that creates an anastomosis between the lacrimal sac mucosa and the nasal mucosa decreases the formation of granulation tissue. But, there is no significant difference of success rate between two groups. PMID- 19297109 TI - Assessing binaural/bimodal advantages using auditory event-related potentials in subjects with cochlear implants. AB - OBJECTIVE: To assess the advantage of binaural and bimodal hearing for subjects with cochlear implant (CI) using auditory event-related potentials as well as speech perception tests. SUBJECTS AND METHODS: The subjects comprised four binaural CI users (CI/CI group) and eleven bimodal CI users, who wore a hearing aid (HA) contralaterally (CI/HA group). All subjects had used binaural or bimodal fitting for over 6 months. Their speech perception was examined in a quiet environment using monosyllabic words. Late cortical waves were measured while subjects were engaged in an oddball task of 1kHz frequent and 2kHz rare tone stimuli. The latencies of event-related potential (N1, N2, P3) were compared for monaural, binaural, and bimodal hearing conditions. RESULTS: Significantly (p<0.01, paired t-test) better speech perception for monosyllabic words was found for both binaural and bimodal hearing than monaural hearing. The latency of N1 did not significantly change for either binaural or bimodal hearing. On the other hand, the latency of N2 was significantly (p<0.01, paired t-test) shorter for binaural and bimodal hearing than for monaural hearing. The latency of P3 was shorter for binaural and bimodal hearing than monaural hearing in all subjects, and the difference was statistically significant in both CI/CI and CI/HA groups (p<0.01, paired t-test). CONCLUSIONS: Better speech perception was obtained for binaural and bimodal hearing than for monaural hearing in CI subjects. The results obtained in the comparison of P3 latency agreed with that of speech perception. Thus, using bilateral hearing devices is recommended for CI subjects. We also found that event-related potentials were useful as an objective tool to assess the advantage of binaural and bimodal hearing for CI subjects. PMID- 19297110 TI - [Drug related deaths in France. A critical view]. AB - BACKGROUND: Available data suggest a reduction in the number of overdoses due to psychoactive substances (tobacco and alcohol excluded) in France. Nevertheless, official levels remain considerably low compared to other European countries, and strong differences between the different sources providing information support the hypothesis of under-reporting. METHODS: Two cross-tabulations of the main sources of information (police data and national mortality registry on one hand; police data and on another hand) allow the use of easy-to-compute, capture recapture based indicators to estimate the invisible population (i.e. hidden overdoses). RESULTS: Results support a significant under-reporting of overdoses, reaching a minimum level of 30%. CONCLUSION: Unlike official claims, there has been a rise in overdoses in France during the last decade. Besides the applied corrections, levels of overdoses still remain low. Attention should be paid to death certification. Other issues of interest are the lack of a common definition and the need for greater institutional coordination. PMID- 19297111 TI - Cerebral venous thrombosis-A pictorial review. AB - Cerebral venous thrombosis is a relatively uncommon but serious neurological condition. It can have disastrous consequences if not diagnosed and treated promptly. It can be missed on initial presentation clinically as well as radiologically unless there is high level of suspicion. This pictorial review is intended to bring an increased awareness among radiology and medical trainees who are involved in the management of these patients. In this review we also discuss the various radiological investigations that can be performed for making the diagnosis and emphasise ways to avoid pitfalls. PMID- 19297112 TI - Determination of short-chain fatty acids in dietary fiber extracts using ion exclusion chromatography with suppressed conductivity detection. AB - A new chromatographic method for the sequential determination of short-chain fatty acids is described. Acetic, propionic and butyric acids were determined in dietary fiber extracts using ion-exclusion chromatography equipped with inverse chemical suppression and conductivity detection. The best optimization of the chromatographic conditions were achieved when a 100 mm x 7.8 mm ion-exclusion column with a solution of 0.5 mmol L(-1) sulfuric acid as eluent in a flow rate of 0.6 mL min(-1) were employed. The organic acids were sequentially separated in less than 10 min with limits of detection ranging from 1 up to 7.5 micromol L(-1) and limits of quantification from 5 up to 25 micromol L(-1). The linearity of the analytical response was studied in the range of 0.005-10 mmol L(-1) for acetic acid and 0.025-10 mmol L(-1) for propionic and butyric acids with coefficients of determination (R(2)) ranging between 0.9985 and 0.9999. The method was tested and proved to be selective, precise, accurate, reproducible and highly sensitive. Finally, the method was applied in the analysis of biological samples. PMID- 19297113 TI - Development of a dual-analyte fluorescent sensor for the determination of bioactive nitrite and selenite in water samples. AB - Nitrite and selenium are two bioactive compounds found in the environment which show beneficial effects for health at low levels but have toxic effects at higher doses. Consequently, quantification of both analytes in water samples results of great interest in areas such as biomedicine, food technology and environmental analysis. In a recent paper, we immobilized the inclusion complex formed between 2,3-diaminonaphthalene (DAN) and 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) in a sol-gel matrix, in order to prepare a highly sensitive reagentless fluorescence-based sensor for the specific measurement of nitrite. Here we have explored the possibility of using the sol-gel immobilized complex to quantify selenite (Se (IV)), the more toxic form of selenium, as well as to act as a dual analyte chemical sensor for simultaneous quantification of both nitrite and selenite in aqueous samples. Results show that (a) inclusion of DAN in HP-beta-CD and its subsequent immobilization in a sol-gel matrix do not modify the reactivity of DAN against selenite, (b) the reaction product formed (4,5 benzopiazselenol) remains into the cyclodextrin increasing considerably its fluorescence quantum yield and avoiding, therefore, its extraction into organic solvents, (c) the developed sensor can detect selenite concentrations at submicromolar level with a minimum detection limit of 13 nM, (d) the immobilized system is able to simultaneously quantify nitrite and selenite at submicromolar concentrations in natural water samples with no further sample pre-treatment. PMID- 19297114 TI - Simultaneous determination of five anthraquinones in medicinal plants and pharmaceutical preparations by HPLC with fluorescence detection. AB - A reversed-phase high-performance liquid chromatography (RP-HPLC) method with fluorescence detection for simultaneous determination of five anthraquinones in Rhubarb collected from nine different locations in China, Polygonum cuspidatum, Polygoni multiflori and three pharmaceutical preparations is proposed and validated. Chromatography was carried out at 25 degrees C on a Hypersil C18 column with the isocratic mobile phase of methanol-0.1% aqueous formic acid (85:15, v/v) at a flow rate of 1.0 ml/min. The excitation and emission wavelengths were set at 440 and 540 nm, respectively. A comprehensive validation of the method included tests of sensitivity, linearity, precision and accuracy. The linear regressions were acquired with r>0.999. Satisfactory intra- and inter day precisions were achieved with R.S.D.s less than 3.95% and the average recovery factors obtained were in the range of 93.2-103.8%. PMID- 19297115 TI - Feasibility of the preoperative Mallampati airway assessment in emergency department patients. AB - BACKGROUND: Multiple predictors have been proposed to assist in identifying patient features that would predict difficult airway management. The Mallampati score (MS) has been shown to be useful in the preoperative assessment of patients being intubated in the operating room. OBJECTIVE: We sought to define the feasibility of this assessment in the Emergency Department. METHODS: A prospective, observational study was performed on all patients being intubated at a university Level I trauma center over a period of 6 months. We recorded and calculated the proportion of patients who were successfully assessed using the MS. Reasons given by individual intubators for failure to assess were recorded. We also tracked patient characteristics between groups and complication rates. RESULTS: Of 328 patients, 32 (10%) were excluded due to incomplete data. Among the remaining 296, 58% were intubated for non-trauma indications, 70% were male, and the mean age was 45.9 years. Only 76 of 296 (26%) (95% confidence interval 21 31%) were able to have the MS performed. Lack of patient cooperation and clinical instability were listed as factors that precluded evaluation in patients whose assessment was unsuccessful. The frequency of procedure-related minor events did not differ significantly between the assessed and non-assessed groups. Major events included two cricothyrotomies in the non-assessed group. CONCLUSIONS: We were unable to perform a Mallampati assessment in three-quarters of our patients requiring emergency intubation. These findings call into question the feasibility of the standard Mallampati assessment in the practice of Emergency Medicine. PMID- 19297116 TI - Using computer agents to explain medical documents to patients with low health literacy. AB - OBJECTIVE: Patients are commonly presented with complex documents that they have difficulty understanding. The objective of this study was to design and evaluate an animated computer agent to explain research consent forms to potential research participants. METHODS: Subjects were invited to participate in a simulated consent process for a study involving a genetic repository. Explanation of the research consent form by the computer agent was compared to explanation by a human and a self-study condition in a randomized trial. Responses were compared according to level of health literacy. RESULTS: Participants were most satisfied with the consent process and most likely to sign the consent form when it was explained by the computer agent, regardless of health literacy level. Participants with adequate health literacy demonstrated the highest level of comprehension with the computer agent-based explanation compared to the other two conditions. However, participants with limited health literacy showed poor comprehension levels in all three conditions. Participants with limited health literacy reported several reasons, such as lack of time constraints, ability to re-ask questions, and lack of bias, for preferring the computer agent-based explanation over a human-based one. CONCLUSION: Animated computer agents can perform as well as or better than humans in the administration of informed consent. PRACTICE IMPLICATIONS: Animated computer agents represent a viable method for explaining health documents to patients. PMID- 19297117 TI - A cognitive behavior therapy-based intervention among poorly controlled adult type 1 diabetes patients--a randomized controlled trial. AB - OBJECTIVE: To examine the impact of a Cognitive Behavior Therapy (CBT)-based intervention on HbA(1c), self-care behaviors and psychosocial factors among poorly controlled adult type 1 diabetes patients. METHODS: Ninety-four type 1 diabetes patients were randomly assigned to either an intervention group or a control group. The intervention was based on CBT and was mainly delivered in group format, but individual sessions were also included. All subjects were provided with a continuous glucose monitoring system (CGMS) during two 3-day periods. HbA(1c), self-care behaviors and psychosocial factors were measured up to 48 weeks. RESULTS: Significant differences were observed with respect to HbA(1c) (P<0.05), well-being (P<0.05), diabetes-related distress (P<0.01), frequency of blood glucose testing (P<0.05), avoidance of hypoglycemia (P<0.01), perceived stress (P<0.05), anxiety (P<0.05) and depression (P<0.05), all of which showed greater improvement in the intervention group compared with the control group. A significant difference (P<0.05) was registered with respect to non severe hypoglycemia, which yielded a higher score in the intervention group. CONCLUSION: This CBT-based intervention appears to be a promising approach to diabetes self-management. PRACTICE IMPLICATIONS: Diabetes care may benefit from applying tools commonly used in CBT. For further scientific evaluation in clinical practice, there is a need for specially educated diabetes care teams, trained in the current approach, as well as cooperation between diabetes care teams and psychologists trained in CBT. PMID- 19297118 TI - Consideration of anatomical structures relevant to the surgical strategy for managing gallbladder carcinoma. AB - AIMS: Gallbladder carcinoma usually presents late with advanced disease. It develops in an anatomically complex area. Consideration is given to resection of relevant local structures with respect to outcome. METHODS: A comprehensive literature review was performed, searching Medline for articles published since 2000, using the MeSH heading of 'gallbladder cancer' and 'surgery'. Abstracts were reviewed and articles retrieved if the main focus of the article centred on the surgical management of gallbladder carcinoma. OBSERVATIONS: Hepatic resection is advocated and tailored to pathological T stage. Lymph node dissection and bile duct resection, as well as en bloc resection of other viscera, remain areas of controversy. CONCLUSIONS: Eastern and Western practice standards of care differ, but hepatic resection with some lymph node dissection is present in both approaches. Philosophy regarding aggression with respect to en bloc resection of adjacent organs and actual extent of lymphatic resection remains disparate. PMID- 19297119 TI - [A tool to evaluate preoperative anxiety in the operating room]. PMID- 19297120 TI - [Plasma exchange therapy in a severe snake bite victim]. PMID- 19297121 TI - [Effect of local anesthetics on the postoperative inflammatory response]. AB - Current knowledge suggests that peripheral inflammation following surgery activates and sensitizes both peripheral and central nervous system. These phenomena involved in the maintenance of the inflammatory response lead to hypersensibility, hyperalgesia and allodynia. Hyperalgesia participates in the general experience of postoperative pain and ALo in the development of chronic pain. A correlation between the ability of treatments to reduce areas of hypersensitivity surrounding the wound after surgery and their ability to reduce the incidence of chronic pain has been shown. For a long time, local anaesthetics have been used for their capacity to block nociceptive input. They can ALo modulate the inflammatory response following a surgical trauma. By inhibiting the nervous conductivity at the site of the trauma, local anesthetics attenuate the sensitization of the nervous system and therefore the inflammatory phenomena. They ALo exert intrinsic anti-inflammatory properties by modulating the local and systemic liberation of inflammatory mediators. The mechanisms involved are not clearly elucidated. Local, systemic, and spinal inflammatory mechanisms may be influenced by local anesthetics through multiple different mechanisms. The therapeutic implications of effects of local anesthetics on local, systemic, and spinal inflammatory responses merit further study. PMID- 19297122 TI - [Learned societies and end of life...]. PMID- 19297123 TI - [Which strategies to use to decrease the risk of pulmonary gastric fluid aspiration during anaesthesia?]. PMID- 19297124 TI - [Resistant hypertension: evaluation and treatment]. AB - Treatment resistant hypertension is defined as a blood pressure not achieving a goal blood pressure (< 140/90 mm Hg). The diagnosis of resistant hypertension requires use of good blood pressure technique to confirm persistently elevated blood pressure levels. Pseudoresistance, including lack of blood pressure control secondary to poor medication adherence or white coat hypertension, must be excluded. The evaluation of patients with resistant hypertension is focused on identifying contributing and secondary causes of hypertension which are guided by the clinical feature of hypertension: metabolic (obstructive sleep apnea, kidney disease), vascular (renal artery atheroma stenosis), endocrine (hyperaldosteronism), familial (renal artery fibrodyspalsia, adrenal causes). Treatment includes removal of contributing factors, appropriate management of secondary causes, and use of effective multidrug regimens. Three antihypertensive medications including ARB or ACEI in addition to calcium channel blocker and to thiazide diuretics is able to control 75% of hypertensive subjects when prescribed in effective doses. The addition of low dose spironolactone to this triple treatment induces significant BP reduction in most patients with resistant hypertension. PMID- 19297125 TI - [Diagnosing the cause of acute dyspnea in elderly patients: role of biomarkers in emergencies]. AB - Acute dyspnea is one of the leading causes of emergency hospitalization of elderly patients. Clinical diagnostic procedures are difficult in this geriatric population. Acute heart failure is the most frequent cause of acute dyspnea in geriatric patients. The use of plasma B natriuretic peptide (BNP) assays in the general population has profoundly improved its medical management. There has also been progress recently for other frequent causes of dyspnea in the elderly, including infection and venous thromboembolic disease. Procalcitonin assays may be useful as a prognostic factor for infectious disease. Nevertheless, the real value of BNP assays in geriatric populations must be clarified by interventional studies. PMID- 19297126 TI - [Anti-CD20 in hematologic oncology]. AB - Twelve years ago, rituximab, a chimeric monoclonal anti-CD20 antibody, became available for the treatment of relapsing follicular lymphoma. It has resulted in an improvement of approximately 20% in survival for nearly all B cell lymphomas. In follicular lymphoma, rituximab combined with chemotherapy is indicated as first-line treatment when treatment is necessary. It is also an interesting option, alone or in combination with chemotherapy for the treatment of relapses, after verifying the persistence of CD20 expression on the surface of malignant cells. As a maintenance treatment after chemotherapy with or without rituximab, it improves relapse-free survival and overall survival. The standard treatment for high-grade diffuse lymphoma, regardless of age and regardless of predicted severity, is now the combination of CHOP chemotherapy and rituximab. The combination of fludarabine, cyclophosphamide, and rituximab is currently the reference treatment for chronic lymphoid leukemia as first-line treatment for patients with few comorbidities and for those with relapses. The role of this antibody in the treatment of more serious forms of lymphoma is under evaluation. The administration of rituximab, alone or in combination, in the treatment of low grade non-follicular lymphoma must be decided as a function of existing protocols in multidisciplinary consultations. New-generation anti-CD20 antibodies, designed to improve still further the already remarkable efficacy/toxicity ratio, are now taking the road that rituximab opened. PMID- 19297127 TI - [Treatment of systemic autoimmune and inflammatory diseases with rituximab]. AB - Rituximab is a chimeric monoclonal antibody that targets CD20 antigen at the surface of B lymphocytes. The efficacy of rituximab in patients with rheumatoid arthritis has been demonstrated in 3 randomized controlled trials. Rituximab is now used in a wide range of systemic autoimmune and inflammatory diseases, as it is well tolerated and efficient. Adverse events are scarce, consisting mainly in reactions during infusion and infectious complications that are favoured by the association of rituximab therapy with other immunosuppressants. Relapses of the disease are common around six months after rituximab infusion. The response to retreatment with rituximab is usually the same that was obtained after the first course of treatment. PMID- 19297128 TI - Comorbidity of motor and language impairments in preschool children of Taiwan. AB - Comorbidity of motor and speech/language impairments was investigated in 363 preschool children between the ages of 5 and 6 years (boys: 205, age 6.04+/-0.48 years; girls: 158, age 5.98+/-0.53 years). The children were sampled from two municipals of Taiwan, and were determined to present no apparent neurological, musculoskeletal, cardiopulmonary system impairment or mental insufficiency. They were administered with three speech/language tests and a motor test (Movement Assessment Battery for Children, or M-ABC). The results showed a significant correlation between the total score of the motor test and the total score of each of the speech and language tests. Regression analysis that controlled for IQ (C TONI) further showed that manual dexterity, but not ball skills or balance, of M ABC was predictive of all scores on the speech and language tests. To determine a deficit on a test, a score at or below the 10th percentile of the norm or a score at or below 1.25SD from the group mean was established as the cutoff. For the speech/language impairment, a deficit on at least two out of the three tests also applied. Following these criteria, 22 children (6.1%) were identified to have Developmental Speech and Language Disorder (DSLD), and 45 (12.4%) to have Developmental Coordination Disorder (DCD). Comorbid DSLD and DCD were found in six children (1.65%). Chi-square analysis revealed a significant correlation between DSLD and DCD (p<.03). The odds of DSLD was higher (by about three-fold) among the children with DCD than among the children without (0.15 vs. 0.05). Comorbid motor and speech/language impairments in preschool children appear to be a significant clinical condition that requires the attention of the therapeutic community. Manual dexterity, in particular, seems to be an important clue for understanding the shared mechanism of motor and speech/language impairments. PMID- 19297129 TI - Rasch analysis of the Developmental Test of Visual-Motor Integration in children with intellectual disabilities. AB - The purpose of this study was to examine the measurement properties of the Developmental Test of Visual-Motor Integration (VMI) in children with intellectual disabilities (ID) ages 4-12 years using the dichotomous Rasch model. The VMI was administered individually to 454 children with ID. Rasch analysis was applied to investigate unidimensionality, item fit to the model, differential item functioning (DIF), and item targeting. Discriminative validity was obtained by receiver operating characteristic (ROC) curve analysis. Items were eliminated if the task was too easy or too difficult, or showed misfit to the Rasch model. The remaining items fitted the unidimensional construct the test was intended to measure and were free of DIF. The Rasch reduced version of the VMI with 9 items appeared to be suited to measure mild degrees of perceptual-motor impairment and demonstrated excellent reliability (0.91). VMI-9 had a larger area under the ROC curve in its ability to differentiate mild versus moderate to severe ID compared with the original version. Taken together, the VMI-9 provides a quick, reliable and valid measure for screening and identifying perceptual-motor deficits in children with ID. PMID- 19297130 TI - From facial emotional recognition abilities to emotional attribution: a study in Down syndrome. AB - Facial expression processing and the attribution of facial emotions to a context were investigated in adults with Down syndrome (DS) in two experiments. Their performances were compared with those of a child control group matched for receptive vocabulary. The ability to process faces without emotional content was controlled for, and no differences appeared between the two groups. Specific impairments were found in the DS group according to the task modalities and the type of facial emotional expressions. In the emotion matching condition, the DS adults showed overall difficulties whereas in the identification and recognition conditions they were particularly impaired when processing the neutral expression. In the emotion attribution task, they exhibited difficulties with the sad expression only and the analysis of their error pattern revealed that they rarely selected this expression throughout the task. The sad emotion was the only one that showed a significant relationship with the facial expression processing tasks. PMID- 19297131 TI - Is there a role for enucleation in the management of ameloblastoma? AB - This study aimed to determine the appropriate long-term management for ameloblastoma and the role of enucleation in the management of the subtypes of ameloblastoma (solid ameloblastomas, cystic ameloblastomas and peripheral ameloblastomas). They differ in their degrees of aggressive behavior and recurrence rates. This is an evidence-based study with review of relevant articles from PubMed, EMBASE and the Cochrane Library. Articles were categorized for quality according to the Oxford Center of Evidence-Based Medicine (CEBM). 58 articles met the inclusion criteria; their evidence level varied from IIA to V. No randomized control trials were identified. Solid and multicystic ameloblastomas have a high recurrence rate (60-80%) with simple enucleation and require more aggressive treatment. The treatment of choice is resection with 1-cm margins. This may require segmental resection in the mandible, and partial maxillectomy in the maxilla. For the unicystic ameloblastoma recurrence rates are high for simple enucleation. The intraluminal subtype of unicystic ameloblastoma may do well with enucleation, but the intramural subtype may not, and since these cannot be identified preoperatively more aggressive treatment is recommended, including peripheral ostectomy or enucleation with subsequent treatment of the surrounding bone with liquid nitrogen, Carnoy's solution, or similar physicochemical modality. The peripheral ameloblastoma has a different origin and responds to local excision. PMID- 19297132 TI - Repeat magnetoencephalography and surgeries to eliminate atonic seizures of non lesional frontal lobe epilepsy. AB - A 7-year-old boy presented with intractable head nodding and atonic seizures since the age of 3 years and severe behavior problems unsuitable for video EEG monitoring. We performed magnetoencephalography (MEG) 4 times: before (1) corpus callosotomy, (2) left frontal cortical resection with multiple subpial transection and (3) resection of residual MEG spike sources, which eliminated seizures. Repeat MEGs consistently localized the epileptogenic zone in a patient with intractable residual non-lesional left frontal lobe epilepsy. PMID- 19297133 TI - Pathways to functional outcomes in schizophrenia: the role of premorbid functioning, negative symptoms and intelligence. AB - BACKGROUND: Social and intellectual premorbid functioning are generally estimated retrospectively, and related to clinical or hospitalization outcomes in schizophrenia. Yet the relationship between premorbid functioning assessed prior to psychiatric hospitalization and postmorbid functional outcomes has not been examined. OBJECTIVES: To test competing models of the relationship between (a) functional outcomes with (b) premorbid functioning assessed on nationally administered tests prior to psychiatric hospitalization, postmorbid intellectual functioning and symptomatology using a historical prospective design. METHODS: Ninety one inpatient and outpatient males with schizophrenia or schizoaffective disorder, aged 19 to 35, were examined using the Positive and Negative Syndrome Scale, the WAIS-III and Strauss and Carpenter social and occupational functional outcome scale. Premorbid intelligence and social functioning data were obtained from national standardized tests administered during high school prior to first hospitalization for schizophrenia. RESULTS: Path modeling showed that premorbid intelligence and behavioral functioning directly predicted postmorbid IQ and negative symptoms, and indirectly predicted postmorbid social and occupational functioning via negative symptoms. Item level analysis indicated that better social and occupational outcomes occurred in a group with few negative symptoms. CONCLUSIONS: Premorbid functioning, postmorbid IQ and negative symptoms are related, yet the relationship between premorbid functioning and postmorbid functional outcomes appears to be mediated by postmorbid negative symptoms. PMID- 19297134 TI - Risk factors and clinical features of infections caused by plasmid-mediated AmpC beta-lactamase-producing Enterobacteriaceae. AB - A case-control study was performed with the objective of analysing risk factors and clinical features of infections caused by plasmid-mediated AmpC beta lactamase (plasmid AmpC)-producing Enterobacteriaceae. All patients infected with plasmid AmpC-producing Enterobacteriaceae in two tertiary care hospitals from December 2006 to August 2007 were included. Plasmid AmpC enzymes were characterised by isoelectric focusing, enzyme inhibition assay and enzyme specific polymerase chain reaction. A total of 30 patients (20 with Klebsiella pneumoniae and 10 with Escherichia coli) were recruited prospectively. CMY-2 and DHA-1 were the most common plasmid AmpC in E. coli and K. pneumoniae, respectively. An independent risk factor for infection with plasmid AmpC producing Enterobacteriaceae was the use of an oxyimino-cephalosporin within 1 month of plasmid AmpC infection [adjusted odds ratio (aOR), 10.8, 95% confidence interval (CI), 1.6-75.4; P=0.016], with the use of a urinary catheter showing borderline significance (aOR, 6, 95% CI 0.93-38.4; P=0.06). An independent risk factor for treatment failure at 72 h was infection due to plasmid AmpC-producing Enterobacteriaceae (aOR, 9.78, 95% CI 1.34-71.17; P=0.02). These results suggest that infections caused by plasmid AmpC-producing isolates significantly increase treatment failure at 72 h and that prior use of an oxyimino-cephalosporin is a risk factor for infections caused by plasmid AmpC-producing Enterobacteriaceae. PMID- 19297136 TI - TRAPDOR double-resonance and high-resolution MAS NMR for structural and template studies in zeolite ZSM-5. AB - A combination of (27)Al magic-angle spinning (MAS)/multiple-quantum (MQ) MAS, and (27)Al-{(14)N} TRAnsfer of Population in DOuble-Resonance (TRAPDOR) nuclear magnetic resonance (NMR) was used to study aluminium environments in zeolite ZSM 5. (27)Al-{(14)N} TRAPDOR experiments, in combination with (14)N NMR were employed to show that the two tetrahedral peaks observed in the (27)Al MAS/3Q-MAS spectra of as-synthesized ZSM-5 are due to aluminium atoms occupying crystallographically inequivalent T-sites. A (13)C-{(27)Al} TRAPDOR experiment was used to study the template, tetrapropyl ammonium bromide (TPABr), in the three-dimensional pore system of ZSM-5. The inequivalency of the methyl groups of TPA was observed in the (13)C-{(27)Al} TRAPDOR spectra of as-synthesized ZSM-5 and the motion of the methyl end of the propyl chain appeared to be more restricted in the sinusoidal channel than in the straight channel. PMID- 19297135 TI - White matter abnormalities and neurocognitive deficits associated with the passivity phenomenon in schizophrenia: a diffusion tensor imaging study. AB - The passivity phenomenon is a distressing Schneiderian first rank symptom in patients with schizophrenia. Based on extant data of functional and structural cerebral changes underlying passivity, we sought to examine cerebral white matter integrity in our subjects. We hypothesised that the passivity phenomenon would be associated with white matter changes in specific cortical (frontal, parietal cortices, and cingulate gyrus) and subcortical regions (thalamus and basal ganglia) and correlated with relevant neurocognitive deficits, compared with characteristics in those without the passivity phenomenon. Thirty-six subjects (11 with passivity and 25 without passivity) with schizophrenia were compared with 32 age-, gender- and handedness-matched healthy controls using diffusion tensor imaging. Neuropsychological testing was administered. Patients with passivity were associated with increased fractional anisotropy within the frontal cortex, cingulate gyrus, and basal ganglia and decreased fractional anisotropy within the thalamus when compared with patients without passivity. Within patients with passivity, fractional anisotropy in the frontal cortex correlated with the age of onset of illness and neurocognitive deficits related to attention and executive functioning. The findings suggest distributed involvement of cortical and subcortical regions underlying passivity and support the notion of neural network models underlying specific psychiatric symptoms such as passivity. PMID- 19297137 TI - Morphology and ciliary pattern of some rare haptorid ciliates, with a description of the new family Kamburophryidae (Protists, Haptoria). AB - We investigated three rare haptorid ciliates, viz., Lagynophrya gibba Kahl (1935), Enchelys lajacola nov. spec., and Spathidium implicatum Kahl (1930), using live observation, silver impregnation, morphometry, and scanning electron microscopy. Lagynophrya gibba, which was rediscovered in peatland soil from Iceland, is referred to a new genus, Kamburophrys, and a new family, Kamburophryidae, based on a unique organelle, the brush membranoid. This structure is near the dorsal brush and composed of very narrowly spaced cilia, about 5microm long. The genus Kamburophrys has a unique combination of features, viz., an oral cone on the oral bulge, an oblique circumoral kinety, and a subapical hump carrying the three-rowed dorsal brush and the brush membranoid. The Kamburophryidae possibly belong to the order Spathidiida. Enchelys lajacola was discovered in mud from granitic rock-pools (Lajas) in Venezuela, South America. The new species is characterized by a bottle-like shape, a macronucleus with the shape of a curved strand, a heterostichad dorsal brush, and rod-shaped toxicysts. Spathidium implicatum, which was rediscovered in an ephemeral meadow puddle near Salzburg city centre, is neotypified and referred to the genus Apertospathula because it has an open circumoral kinety. PMID- 19297138 TI - Urban air pollution: influences on olfactory function and pathology in exposed children and young adults. AB - Mexico City (MC) residents are exposed to severe air pollution and exhibit olfactory bulb inflammation. We compared the olfactory function of individuals living under conditions of extreme air pollution to that of controls from a relatively clean environment and explore associations between olfaction scores, apolipoprotein E (APOE) status, and pollution exposure. The olfactory bulbs (OBs) of 35 MC and 9 controls 20.8+/-8.5 years were assessed by light and electron microscopy. The University of Pennsylvania Smell Identification Test (UPSIT) was administered to 62 MC/25 controls 21.2+/-2.7 years. MC subjects had significantly lower UPSIT scores: 34.24+/-0.42 versus controls 35.76+/-0.40, p=0.03. Olfaction deficits were present in 35.5% MC and 12% of controls. MC APOE epsilon 4 carriers failed 2.4+/-0.54 items in the 10-item smell identification scale from the UPSIT related to Alzheimer's disease, while APOE 2/3 and 3/3 subjects failed 1.36+/ 0.16 items, p=0.01. MC residents exhibited OB endothelial hyperplasia, neuronal accumulation of particles (2/35), and immunoreactivity to beta amyloid betaA(42) (29/35) and/or alpha-synuclein (4/35) in neurons, glial cells and/or blood vessels. Ultrafine particles were present in OBs endothelial cytoplasm and basement membranes. Control OBs were unremarkable. Air pollution exposure is associated with olfactory dysfunction and OB pathology, APOE 4 may confer greater susceptibility to such abnormalities, and ultrafine particles could play a key role in the OB pathology. This study contributes to our understanding of the influences of air pollution on olfaction and its potential contribution to neurodegeneration. PMID- 19297139 TI - Influence of water-soluble flavonoids, quercetin-5'-sulfonic acid sodium salt and morin-5'-sulfonic acid sodium salt, on antioxidant parameters in the subacute cadmium intoxication mouse model. AB - Water-soluble quercetin-5'-sulfonic acid sodium salt (NaQSA) and morin-5' sulfonic acid sodium salt (NaMSA) could exert an antagonistic effect on cadmium intoxication. The aim of the study was to examine the influence of these substances on superoxide dismutase (SOD) and glutathione (GSH) levels in the mouse liver in the subacute cadmium intoxication model. NaQSA and NaMSA significantly counteracted cadmium-induced decreases in SOD and GSH levels. No significant differences in SOD and GSH levels between groups exposed to cadmium receiving NaQSA or/and NaMSA were observed. PMID- 19297140 TI - Antigen choice in adoptive T-cell therapy of cancer. AB - Immunotherapy of cancer through adoptive transfer of genetically engineered T cells constitutes a more powerful strategy than attempts to mobilize the endogenous T-cell repertoire. Application of this technology in patients offers great opportunities towards a long-awaited breakthrough in cancer immunotherapy. However, recent findings in preclinical mouse models indicate that infusion of T cells directed against tumor-associated auto-antigens can be associated with higher 'on target' toxicity than was anticipated on the basis of anti-tumor vaccination studies. Critical evaluation of candidate target antigens is required to ensure that T-cell receptor gene therapy will result in preferential attack of tumor cells in the absence of irreversible damage to vital somatic tissues. PMID- 19297141 TI - Similar efficacy of ciclesonide versus prednisolone to treat asthma worsening after steroid tapering. AB - RATIONALE: Oral corticosteroids effectively treat asthma exacerbations but are associated with well-described side effects. OBJECTIVE: This study compared the efficacy and safety of a high dose of an inhaled corticosteroid with oral prednisolone in patients with worsening of their asthma after medication withdrawal. METHODS: Patients tapered off their inhaled corticosteroids until they reached predefined criteria of "worsening asthma". Randomized patients (n=130) were treated double blind with either ciclesonide 800mug twice daily (starting with 800mug hourly for 3h after randomization) or prednisolone 40mg once daily for 2 weeks. Spirometry, daily asthma symptoms, morning and evening peak expiratory flow and blood parameters were assessed in all, methacholine challenge and inflammatory measures were determined in induced sputum in a subset of patients. RESULTS: Ciclesonide was as effective as prednisolone in improving forced expiratory flow in 1s, morning peak expiratory flow and symptoms, the latter improving more rapidly with ciclesonide. No differences were found in methacholine responsiveness or inflammatory measures in sputum or blood. Ciclesonide caused significantly less reduction in morning plasma cortisol levels (p<0.0001). CONCLUSION: This study shows that inhaled ciclesonide (800mug twice daily) has comparable efficacy to oral prednisolone (40mg once daily) to regain asthma control in patients with asthma worsening. The more rapid onset and smaller effect on cortisol suppression suggest a better safety profile of ciclesonide. PMID- 19297142 TI - Impact of nitrate-enhanced leachate recirculation on gaseous releases from a landfill bioreactor cell. AB - This study evaluates the impact of nitrate injection on a full scale landfill bioreactor through the monitoring of gaseous releases and particularly N(2)O emissions. During several weeks, we monitored gas concentrations in the landfill gas collection system as well as surface gas releases with a series of seven static chambers. These devices were directly connected to a gas chromatograph coupled to a flame ionisation detector and an electron capture detector (GC FID/ECD) placed directly on the field. Measurements were performed before, during and after recirculation of raw leachate and nitrate-enhanced leachate. Raw leachate recirculation did not have a significant effect on the biogas concentrations (CO(2), CH(4) and N(2)O) in the gas extraction network. However, nitrate-enhanced leachate recirculation induced a marked increase of the N(2)O concentrations in the gas collected from the recirculation trench (100-fold increase from 0.2 ppm to 23 ppm). In the common gas collection system however, this N(2)O increase was no more detectable because of dilution by gas coming from other cells or ambient air intrusion. Surface releases through the temporary cover were characterized by a large spatial and temporal variability. One automated chamber gave limited standard errors over each experimental period for N(2)O releases: 8.1 +/- 0.16 mg m(-2) d(-1) (n = 384), 4.2 +/- 0.14 mg m(-2) d( 1) (n = 132) and 1.9 +/- 0.10 mg m(-2) d(-1) (n = 49), during, after raw leachate and nitrate-enhanced leachate recirculation, respectively. No clear correlation between N(2)O gaseous surface releases and recirculation events were evidenced. Estimated N(2)O fluxes remained in the lower range of what is reported in the literature for landfill covers, even after nitrate injection. PMID- 19297145 TI - Manganese dioxide as an alternative cathodic catalyst to platinum in microbial fuel cells. AB - In this paper, three manganese dioxide materials, alpha-MnO(2), beta-MnO(2), gamma-MnO(2) were tested as alternative cathodic catalysts to platinum (Pt) in air-cathode microbial fuel cells (MFCs). Prepared via hydrothermal method, the manganese dioxides were characterized by X-ray powder diffraction patterns (XRD), the Brunauer-Emmett-Teller (BET) method and their average oxidation states (AOS) were determined by the potential voltammetric titration method. The electro catalytic activity of MnO(2) in neutral pH solution was determined by linear sweep voltammetry (LSV) and the results showed that all manganese dioxides can catalyze oxygen reduction reaction (ORR) in neutral medium with different catalytic activities. beta-MnO(2) appeared to hold the highest catalytic activity due to its highest BET surface area and AOS. Beta-MnO(2) was further used as cathode catalyst in both cube and tube air-cathode MFCs, in which using Klebsiella pneumoniae (K. pneumoniae) biofilm as biocatalyst and utilizing glucose as a substrate in the anode chamber. It was found that tube MFC produced higher output power, with the maximum volumetric power density of 3773+/-347 mW/m(3), than cube MFC. This study suggests that using beta-MnO(2) instead of Pt could potentially improve the feasibility of scaling up MFC designs for real applications by lowering production cost. PMID- 19297144 TI - Structural properties and sensing performance of high-k Nd2TiO5 thin layer-based electrolyte-insulator-semiconductor for pH detection and urea biosensing. AB - For high sensitive pH sensing, an electrolyte-insulator-semiconductor (EIS) device with Nd(2)TiO(5) thin layers fabricated on Si substrates by means of reactive sputtering and the subsequent post-deposition annealing (PDA) treatment was proposed. In this work, the effect of thermal annealing (600, 700, 800, and 900 degrees C) on the structural characteristics of Nd(2)TiO(5) thin layer was investigated by X-ray diffraction, X-ray photoelectron spectroscopy, and atomic force microscopy. The observed structural properties were then correlated with the resulting pH sensing performances. For enzymatic field-effect-transistors based urea biosensing, a hybrid configuration of the proposed Nd(2)TiO(5) thin layer with urease-immobilized alginate film attached was established. Within the experimental conditions investigated, the EIS device with the Nd(2)TiO(5) thin layer annealed at 800 degrees C exhibited a higher pH detection sensitivity of 57.2 mV/pH, a lower hysteresis voltage of 2.33 mV, and a lower drift rate of 1.80 mV/h compared to those at other annealing temperatures. These results are attributed to the formation of a thinner low-k interfacial layer at the oxide/Si interface and the higher surface roughness occurred at this annealing temperature. Furthermore, the presented urea biosensor was also proved to be able to detect urea with good linearity (R(2)=0.99) and reasonable sensitivity of 9.52 mV/mM in the urea concentration range of 3-40 mM. As a whole, the present work has provided some fundamental data for the use of Nd(2)TiO(5) thin layer for EIS based pH detection and the extended application for biosensing. PMID- 19297143 TI - Nanohole arrays of mixed designs and microwriting for simultaneous and multiple protein binding studies. AB - We demonstrate using nanohole arrays of mixed designs and a microwriting process based on dip-pen nanolithography to monitor multiple, different protein binding events simultaneously in real-time based on the intensity of Extraordinary Optical Transmission of nanohole arrays. The microwriting process and small footprint of the individual nanohole arrays enabled us to observe different binding events located only 16 microm apart, achieving high spatial resolution. We also present a novel concept that incorporates nanohole arrays of different designs to improve confidence and accuracy of binding studies. For proof of concept, two types of nanohole arrays, designed to exhibit opposite responses to protein bindings, were fabricated on one transducer. Initial studies indicate that the mixed designs could help to screen out artifacts such as protein intrinsic signals, providing improved accuracy of binding interpretation. PMID- 19297146 TI - Future directions in non-endocrine systemic treatment of metastatic prostate cancer. AB - The management of prostate cancer that has become resistant to androgen deprivation is becoming more complex. Chemotherapy using docetaxel is now an established therapy for such patients with metastatic disease. Other agents including the epothilones are being evaluated and combination regimens have shown significant activity in the phase 2 setting. Other approaches for instance bisphosphonates and small molecule targeted agents are being investigated although their place in therapy is still to be determined. The use of hormones post-chemotherapy may also be useful for a subset of patients. Determining endpoints in phase 2 studies remain a problem as PSA on its own may be unreliable - new guidelines for reporting such studies have just been released which should allow some standardisation in approach. Combining various approaches is likely to depend on the pattern and speed of progression following failure of androgen deprivation. PMID- 19297147 TI - Selective inhibition of methanogenesis to enhance ethanol and n-butyrate production through acetate reduction in mixed culture fermentation. AB - Acetate reduction is an alternative digestion process to convert organic waste into ethanol. Using acetate for fuel ethanol production offers the opportunity to use organic waste materials instead of sugar-containing feedstock. Methanogenesis, however, competes with acetate reduction for acetate and hydrogen and lowers the final efficiency. The aim of this research is to selectively inhibit methanogenesis and to enhance acetate reduction. Acetate reduction was stimulated in batch tests at pH between 4.5 and 8; and at pH 6 with and without thermal pre-treatment. It was found that methanogenesis was selectively inhibited while acetate reduction was enhanced after thermal pre-treatment incubated at pH 6. Initially the acetate reduction yielded 7.7+/-3.2mM ethanol with an efficiency of 60.2+/-8.7%, but later on it was consumed to form 7.02+/-0.85 mM n-butyrate with an efficiency of 76.2+/-14.0%. It was the first time demonstrated that n butyrate can be produced by mixed cultures from only acetate and hydrogen. PMID- 19297148 TI - Enhanced 2,3-butanediol production by Klebsiella oxytoca using a two-stage agitation speed control strategy. AB - Batch fermentative production of 2,3-butanediol by Klebsiella oxytoca was investigated using various oxygen supply methods though varying agitation speed. Based on the analysis of three kinetic parameters including specific cell growth rate (micro), specific glucose consumption rate (q(s)) and specific 2,3 butanediol formation rate (q(p)), a two-stage agitation speed control strategy, aimed at achieving high concentration, high yield and high productivity of 2,3 butanediol, was proposed. At the first 15 h, agitation speed was controlled at 300 rpm to obtain high mu for cell growth, subsequently agitation speed was controlled at 200 rpm to maintain high q(p) for high 2,3-butanediol accumulation. Finally, the maximum concentration of 2,3-butanediol reached 95.5 g l(-1) with the yield of 0.478 g g(-1) and the productivity of 1.71 g l(-1)h(-1), which were 6.23%, 6.22% and 22.14% over the best results controlled by constant agitation speeds. PMID- 19297149 TI - Characterization and fouling properties of exopolysaccharide produced by Klebsiella oxytoca. AB - Klebsiella oxytoca produced a type of exopolysaccharide (EPS) with the average molecular weight (Mw) of 116,018 Da and the average size of 260 nm. The EPS monosaccharide components contained rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose and the molar ratio among them was 0.033:0.0411:0.0147:0.0051:0.2393:0.0986:0.1304. Typical EPS absorption peaks in FT-IR spectrum and pseudoplastic properties were also revealed. The polyvinylidenefluoride (PVDF) membrane showed a relatively larger flux decline resulted from the EPS fouling. The EPS filtration was dominated by more than one mechanism at the beginning phase and mainly by the cake formation at the later phase for both membranes. The pore blocking resistance had a predominant contribution to the filtration resistance and the cake resistance played a secondary role for both the membranes. The EPS adsorption resulted in a weak membrane fouling. The PVDF membrane exhibited a larger adsorption resistance than the polypropylene (PP) membrane. PMID- 19297150 TI - Butyric acid fermentation in a fibrous bed bioreactor with immobilized Clostridium tyrobutyricum from cane molasses. AB - Butyrate fermentation by immobilized Clostridium tyrobutyricum was successfully carried out in a fibrous bed bioreactor using cane molasses. Batch fermentations were conducted to investigate the influence of pH on the metabolism of the strain, and the results showed that the fermentation gave a highest butyrate production of 26.2 g l(-1) with yield of 0.47 g g(-1) and reactor productivity up to 4.13 g l(-1)h(-1) at pH 6.0. When repeated-batch fermentation was carried out, long-term operation with high butyrate yield, volumetric productivity was achieved. Several cane molasses pretreatment techniques were investigated, and it was found that sulfuric acid treatment gave better results regarding butyrate concentration (34.6+/-0.8 g l(-1)), yield (0.58+/-0.01 g g(-1)), and sugar utilization (90.8+/-0.9%). Also, fed-batch fermentation from cane molasses pretreated with sulfuric acid was performed to further increase the concentration of butyrate up to 55.2 g l(-1). PMID- 19297151 TI - Bioconversion of lignocellulosic fraction of water-hyacinth (Eichhornia crassipes) hemicellulose acid hydrolysate to ethanol by Pichia stipitis. AB - Fermentation of acid hydrolysate of water-hyacinth (Eichhornia crassipes), a free floating aquatic plant has been investigated for ethanol production. The dilute acid treatment has been applied to utilize the maximum hemicellulosic content of the water-hyacinth. The goal of this work was to investigate, both experimentally and theoretically using mathematical tools, a fermentative system utilizing water hyacinth (Eichhornia crassipes) hemicellulose acid hydrolysate as a substrate for ethanol production using Pichia stipitis. It was found that 72.83% of xylose was converted to ethanol with a yield of 0.425 g(p)/g(s) and productivity of 0.176 g(p)/L/h. An appropriate mathematical model was developed to explain theoretically the bioconversion of this hemicellulose acid hydrolysate to ethanol and the model was tested statistically to check the validity of the model. PMID- 19297152 TI - Adsorption of chromium (VI) ion from aqueous solution by succinylated mercerized cellulose functionalized with quaternary ammonium groups. AB - Succinylated mercerized cellulose (cell 1) was used to synthesize an anion exchange resin. Cell 1, containing carboxylic acid groups was reacted with triethylenetetramine to introduce amine functionality to this material to obtain cell 2. Cell 2 was reacted with methyl-iodide to quaternize the amine groups from this material to obtain cell 3. Cells 2 and 3 were characterized by mass percent gain, degree of amination and quaternization, FTIR and CHN. Cells 2 and 3 showed degrees of amination and quaternization of 2.8 and 0.9 mmol/g and nitrogen content of 6.07% and 2.13%, respectively. Cell 3 was used for Cr (VI) adsorption studies. Adsorption equilibrium time and optimum pH for Cr (VI) adsorption were found to be 300 min and 3.1, respectively. The Langmuir isotherm was used to model adsorption equilibrium data. The adsorption capacity of cell 3 was found to be 0.829 mmol/g. Kinetic studies showed that the rate of adsorption of Cr (VI) on cell 3 obeyed a pseudo-second-order kinetic model. PMID- 19297153 TI - Chiral N-phosphonyl imine chemistry: asymmetric synthesis of alpha,beta-diamino esters by reacting phosphonyl imines with glycine enolates. AB - Chiral phosphonyl imines attached with N-isopropyl protection group were found to react with lithium glycine enolates under convenient conditions to give alpha,beta-diamino esters. Thirteen examples have been examined in good to excellent chemical yields (85-97%) diastereoselectivity (up to 99% de). By treating with HBr at room temperature, the chiral auxiliary can be readily removed and recycled. The absolute structure has been unambiguously determined by converting a product to a known sample. PMID- 19297155 TI - Synthesis and antiviral activities of novel acylhydrazone derivatives targeting HIV-1 capsid protein. AB - HIV-1 capsid protein (CA) plays important roles in the viral replication cycle. A number of acylhydrazone derivatives that act as inhibitors of HIV-1 CA assembly, were designed and synthesized. The synthesized compounds were tested for their antiviral activities and cytotoxicities using CEM cells. Some derivatives also were assayed for their ability to inhibit HIV-1 CA assembly in vitro. Among them, compounds 14f and 14i display the most promising potency with EC(50) values of 0.21 and 0.17 microM respectively. PMID- 19297154 TI - Discovery of betrixaban (PRT054021), N-(5-chloropyridin-2-yl)-2-(4-(N,N dimethylcarbamimidoyl)benzamido)-5-methoxybenzamide, a highly potent, selective, and orally efficacious factor Xa inhibitor. AB - Systematic SAR studies of in vitro factor Xa inhibitory activity around compound 1 were performed by modifying each of the three phenyl rings. A class of highly potent, selective, efficacious and orally bioavailable direct factor Xa inhibitors was discovered. These compounds were screened in hERG binding assays to examine the effects of substitution groups on the hERG channel affinity. From the leading compounds, betrixaban (compound 11, PRT054021) has been selected as the clinical candidate for development. PMID- 19297156 TI - Exploring the PI3K alpha and gamma binding sites with 2,6-disubstituted isonicotinic derivatives. AB - A homology model of the p110alpha catalytic subunit of PI3Kalpha was generated from the p110gamma crystal structure. Using this model, an isonicotinic scaffold was designed for chemically exploring the PI3Kalpha and gamma binding sites. A focused library of derivatives was synthesized and tested. The morpholine acids 5a and 5b proved to be the most potent analogs. PMID- 19297157 TI - Synthesis, configurational stability and stereochemical biological evaluations of (S)- and (R)-5-hydroxythalidomides. AB - The first asymmetric synthesis of (S)- and (R)-5-hydroxythalidomides, one of thalidomide's major metabolites, was achieved using HMDS/ZnBr(2)-induced imidation as a key reaction. 5-Hydroxythalidomide was found to be configurationally more stable than thalidomide at physiological pH. Stereochemical biological effects of thalidomide and 5-hydroxythalidomide on anti angiogenesis and antitumor activities were also investigated using racemic and pure enantiomers. PMID- 19297158 TI - Anthranilamide-based N,N-dialkylbenzamidines as potent and orally bioavailable factor Xa inhibitors: P4 SAR. AB - Anthranilamide-based benzamidine compound 4 and its N-substituted analogs were designed and examined as factor Xa inhibitors using substituted benzamidines as unconventional S4 binding element. A group of N,N-dialkylbenzamidines (11, 17 and 24) have been discovered as potent factor Xa inhibitors with strong anticoagulant activity and promising oral PK profiles. PMID- 19297159 TI - High-resolution ultrasonographic features of axillary lymph node metastasis in patients with breast cancer. AB - To determine ultrasound (US) features that most accurately predict the presence of axillary lymph node metastasis, we retrospectively analysed the results of preoperative US breast examinations of axillary lymph nodes in 425 consecutive patients who subsequently underwent surgery for invasive breast cancer. We compared the US findings with pathologic results for axillary lymph node metastasis. US features included length of the longest (L) and shortest (S) axes, L/S ratio, cortical thickness, presence of hilum and shape. The results of multivariate logistic regression analysis revealed that cortical thickness greater than 3mm was the most accurate indicator, with 4.14 times increased risk of the presence of an axillary lymph node metastasis as compared to cortical thickness less than 3mm. The absence of a hilum showed the highest specificity for axillary lymph node metastasis (94.6%), but low sensitivity. PMID- 19297160 TI - The trend of pressure ulcer prevalence rates in German hospitals: results of seven cross-sectional studies. AB - Pressure ulcer prevalence rates provide useful information about the magnitude of this health problem. Only limited information on pressure ulcers in Germany was available before 2001. The purpose of this study was to compare results of seven pressure ulcer prevalence surveys which were conducted annually between 2001 and 2007 and to explore whether pressure ulcer prevalence rates decreased. The second aim was to evaluate if the measured prevalence rates of our sample could be generalised for all German hospitals. Results of seven point pressure ulcer prevalence studies conducted in 225 German hospitals were analysed. Chi-square tests, chi-square trend tests and one-way ANOVA to assess differences and trends across the years were applied. The sample was stratified according to pressure ulcer risk and speciality. Finally, study samples were compared with the potential population. In total data of 40,247 hospital patients were analysed. The overall pressure ulcer prevalence rate in German hospitals was 10.2%. Patient samples of each year were comparable regarding gender, age and pressure ulcer risk. Pressure ulcer prevalence rates decreased from 13.9% (year 2001) to 7.3% (year 2007) (p<0.001). When excluding non-blanchable erythema prevalence rates decreased from 6.4% (year 2001) to 3.9% (year 2007) (p=0.015). An explicit decrease of prevalence rates was observed on geriatric wards. Prevalence rates on neurological and intensive care units remained stable. With some limitations our study results are representative for all hospitals within Germany. It is highly probable that the decrease of prevalence rates was due to an increased awareness of the pressure ulcer problem in Germany and subsequent efforts to improve pressure ulcer prevention and treatment. The quality of clinical practice regarding pressure ulcer prevention and treatment has improved. However, pressure ulcers are still relevant and require attention. In 2007, one out of 10 hospital patients who were at pressure ulcer risk had at least one pressure related skin damage. PMID- 19297161 TI - Commentary to: Postural and gait performance in children with attention deficit/hyperactivity disorder [Gait Posture 2008]. PMID- 19297162 TI - Intramedullary subependymoma of the thoracic spinal cord. AB - We report a rare case of a thoracic intramedullary subependymoma in a 37-year-old woman. The patient developed a monoparesis of the right leg after a subtotal resection of the tumor. During the 30 months of follow-up the neurological deficit improved and the patient remained symptom free without progression of the remnant tumor. A complete resection of the tumor is usually curative with improved function. However, aggressive surgery may cause either worsening of an existing deficit or the development of new deficits. In patients with poor delineation between the tumor and the spinal cord, subtotal removal and close follow-up should be considered. PMID- 19297163 TI - Taste perception abnormalities after acute stroke in postmenopausal women. AB - The study aims to elucidate the characteristics of post-stroke taste dysfunction in postmenopausal women. Taste function in 120 consecutive postmenopausal women with acute (<7 days) stroke was compared with that of age-matched control subjects (n=109). The agents used were: sodium chloride for saltiness, sucrose for sweetness, glacial acetic acid for sourness and quinine hemisulfate for bitterness. Detection and recognition thresholds were performed by the three stimulus drop technique. Taste threshold values beyond two standard deviations of normal were considered "abnormal". For postmenopausal women after acute stroke, abnormal detection thresholds for the ability to taste sweetness, saltiness, sourness and bitterness were found in 33%, 21%, 35% and 30% of women, respectively, and abnormal recognition thresholds were found in 40%, 34%, 42% and 33% of women respectively. The taste dysfunction occurred ipsilaterally, contralaterally or bilaterally, and was not related to the side or location of the lesion. Large (>2 cm) lesions were more frequently associated with sweet and salty taste dysfunction than small lesions (p<0.05). Follow-up examination in 23 patients at 24 to 31 months (mean 27 months) after the initial evaluation showed that the taste abnormality persisted in 8 (35%) patients. Taste perception abnormalities are common and often persistent in stroke patients. The dysfunction can occur ipsilaterally, contralaterally or bilaterally. PMID- 19297164 TI - Acute non-traumatic spinal subdural haematoma: an unusual aetiology. AB - We report an unusual case of a spinal subdural haematoma associated with a ruptured spinal aneurysm. The delayed diagnosis or misdiagnosis of this rare entity can have disastrous consequences. We discuss various possible aetiologies and its association with spinal aneurysms. PMID- 19297165 TI - Immediate postoperative death due to hypothalamic injury following surgery for craniopharyngioma. AB - Autonomic disturbances due to hypothalamic injury that result in postoperative death are rare complications following surgery for craniopharyngioma. We discuss the case of a child who died due to hypothalamic injury following radical excision of a multi-compartmental craniopharyngioma. Mechanisms and clinical manifestations of hypothalamic injury and ways to avoid this fatal complication are discussed. PMID- 19297166 TI - A predominantly left upper limb sensory neuronopathy as a manifestation of a metastatic neuroendocrine malignancy. AB - An 83-year-old woman presented with a predominantly left upper limb sensory ganglionopathy. She was found to have metastatic, poorly differentiated, neuroendocrine malignancy. Anti-neuronal antibodies were not detected. PMID- 19297167 TI - The relationship between low bone mineral density and Parkinson's disease in a Korean population. AB - There have been few studies concerning osteoporosis in patients with Parkinson's disease (PD), even though patients with PD have a high incidence of falls. Using a Korean population, we investigated the association between bone mineral density (BMD) and PD. A total of 107 patients with PD were compared with age-matched and sex-matched normal controls. Using the Hoehn & Yahr staging system, we classified the patients with PD into three subgroups. Parts II and III of the Unified Parkinson's Disease Rating Scale were administered to evaluate the relationship between the severity of PD and BMD. In conclusion, in the PD population we found that decreased BMD was related to the severity of immobility of patients, and that BMD changes were greater at the femoral neck than the spine area. Therefore, we assert that clinicians should consider administering screening tests to elderly patients with PD and provide appropriate primary or secondary prophylactic treatment for osteoporosis. PMID- 19297168 TI - Peripheral median nerve stimulation for the treatment of iatrogenic complex regional pain syndrome (CRPS) type II after carpal tunnel surgery. AB - We report on the use and follow-up of direct peripheral nerve stimulation of the median nerve for the treatment of iatrogenic complex regional pain syndrome (CRPS). A 56-year-old woman presented with CRPS type II in the right forearm and hand, which had started after multiple carpal tunnel surgeries and had lasted for 2 years. The visual analogue scale (VAS) score was 8-10 out of 10. After a successful 15-day trial of median nerve peripheral nerve stimulation via a quadripolar lead in the right carpal tunnel space, an implantable pulse generator was inserted in the right infraclavicular space. The VAS score decreased to 1-2 out of 10 and the patient regained the ability to sleep. After 36 months of follow-up, the patient was still experiencing good pain relief without other treatment. We conclude that peripheral nerve stimulation is easy to use in pain management and could offer a valid treatment option for iatrogenic CRPS type II. PMID- 19297169 TI - Sphenoid sinus aspergillosis simulating pituitary tumor in immunocompetent patient. AB - Aspergillosis of the sphenoid sinus is rare in immunocompetent patients. It may be mistaken for a sellar region tumor. A 65-year-old, human immunodeficiency virus-negative man presented with a 3-week history of cranial nerve III paresis and visual deterioration. The patient had a long-term history of tobacco snuff abuse. CT scans and MRI demonstrated a space-occupying lesion of the sellar and sphenoid sinus region. Presumptive diagnosis of pituitary macroadenoma was made and the patient was operated on via a transnasal-transsphenoidal approach. After the sphenoid sinus was opened, a yellow-brownish gluey material with crumbly debris extruded and was aspirated. The dura was intact. Histopathology revealed numerous Aspergillus hyphae without tissue invasion. Postoperatively, the cranial nerve III paresis resolved in a few days and visual acuity improved. Sphenoid sinus aspergillosis should be included in the differential diagnosis of sellar region processes, even in immunocompetent patients. Early diagnosis and transsphenoidal removal provides good results without the need for systemic antifungal therapy in non-invasive aspergillosis. PMID- 19297170 TI - Concussion in sport. PMID- 19297171 TI - Effect of gelatin hydrogel incorporating fibroblast growth factor 2 on human meniscal cells in an organ culture model. AB - Efforts to use growth factors to enhance the healing potential of the meniscus have been impeded because their half-lives are too short to maintain the biological activity. The thread was coated with gelatin hydrogel and fibroblast growth factor 2 (FGF 2) was biologically stabilized by incorporating in a gelatin hydrogel-coated thread. The purpose of this study is to investigate the effect of gelatin hydrogel-coated thread incorporating FGF 2 on human meniscal cells in an organ culture. Twenty-five menisci were cut into small pieces, and selected pieces were sutured with gelatin hydrogel-coated thread incorporating FGF 2 (FGF(+) group) or physiologic saline (FGF(-) group), followed by organ culture. The meniscal samples histologically evaluated 4, 7, and 14 days later. The cell density and the number of PCNA-positive cells for the FGF(+) group were higher than those of the FGF(-) group, while the number of TUNEL-positive cells was lower. These results suggest that FGF 2 stimulates the proliferation of meniscal cells and inhibits meniscal cell death. Gelatin hydrogel-coated threads releasable FGF 2 may be useful to promote repairing of human meniscus. PMID- 19297172 TI - Carbonic anhydrase inhibitors. Inhibition of the beta-class enzymes from the fungal pathogens Candida albicans and Cryptococcus neoformans with aliphatic and aromatic carboxylates. AB - The inhibition of the beta-carbonic anhydrases (CAs, EC 4.2.1.1) from the pathogenic fungi Cryptococcus neoformans (Can2) and Candida albicans (Nce103) with carboxylates such as the C1-C5 aliphatic carboxylates, oxalate, malonate, maleate, malate, pyruvate, lactate, citrate and some benzoates has been investigated. The best Can2 inhibitors were acetate and maleate (K(I)s of 7.3-8.7 microM), whereas formate, acetate, valerate, oxalate, maleate, citrate and 2,3,5,6-tetrafluorobenzoate showed less effective inhibition, with K(I)s in the range of 42.8-88.6 microM. Propionate, butyrate, malonate, L-malate, pyruvate, L lactate and benzoate, were weak Can2 inhibitors, with inhibition constants in the range of 225-1267 microM. Nce103 was more susceptible to inhibition with carboxylates compared to Can2, with the best inhibitors (maleate, benzoate, butyrate and malonate) showing K(I)s in the range of 8.6-26.9 microM. L-Malate and pyruvate together with valerate were the less efficient Nce103 inhibitors (K(I)s of 87.7-94.0 microM), while the remaining carboxylates showed a compact behavior of efficient inhibitors (K(I)s in the range of 35.1-61.6 microM). Notably the inhibition profiles of the two fungal beta-CAs was very different from that of the ubiquitous host enzyme hCA II (belonging to the alpha-CA family), with maleate showing selectivity ratios of 113.6 and 115 for Can2 and Nce103, respectively, over hCA II inhibition. Therefore, maleate is a promising starting lead molecule for the development of better, low nanomolar, selective beta-CA inhibitors. PMID- 19297173 TI - Argifin; efficient solid phase total synthesis and evaluation of analogues of acyclic peptide. AB - An effective solid phase synthesis of Argifin, providing subsequent access to effective synthesis of analogues, was developed in 13% overall yield, as well as elucidating structure-activity relationships. The novel acyclic peptide 18b, prepared from a synthetic intermediate of Argifin, was found to be 70 times more potent as an inhibitor of Serratia marcescens chitinases B than Argifin itself. PMID- 19297174 TI - Synthesis, activity and molecular modeling of a new series of chromones as low molecular weight protein tyrosine phosphatase inhibitors. AB - Protein tyrosine phosphatases (PTP) are crucial elements in eukaryotic signal transduction. Several reports suggested that the LMW-PTP family has oncogenic relevance. Moreover, LMW-PTP has been recognized as a negative regulator of insulin-mediated mitotic and metabolic signaling. Thus, inhibition of the LMW-PTP can be considered an attractive approach for the design of new therapeutic agents for the treatment of type II diabetes and for new antitumoral drugs. To date very few (and weak) inhibitors of LMW-PTP have been identified. On the basis of the reported weak activity of some flavonoids on phosphatases, we discovered a lead that originated a new class of highly active LMW-PTP inhibitors; these compounds inhibit also PTP-1B and are active in cellular assays. Docking experiments and SAR highlighted the possible binding mode of these compounds to the enzyme, putting the background for the future optimization of their inhibitory activity and selectivity towards the closely related enzyme PTP-1B. PMID- 19297175 TI - Validation of rapid methods for the determination of radiostrontium in milk. AB - A rapid method of (89)Sr and (90)Sr comprising ion chromatography for preconcentration and Sr extraction chromatography for separation of Sr from Ca, Ba and Y was validated with spiked milk samples. An (89)Sr/(90)Sr activity ratio of up to 12 showed the relative bias was within +/-20%. The separation time of Sr was 7h and the chemical recovery of Sr ranged from 80% to 95%. The detection limit for 500 mL milk and 90 min counting time was 0.1 Bq L(-1). PMID- 19297176 TI - Recoil effect on beta-decaying in vivo generators, interpreted for (103)Pd/(103m)Rh. AB - The use of Auger emitters as potential radiopharmaceuticals is being increasingly investigated. One of the radionuclides of interest is (103m)Rh, which can be produced from (103)Ru or (103)Pd in an in vivo generator. A potential problem, however, is the recoil of the (103m)Rh out of the carrier molecule and even out of the target cell. In order to determine the likelihood of this happening in the (103)Pd/(103m)Rh, case calculations were made to prove that this does not happen. The equations were generalised for all radionuclides with an atomic mass of 10 240 as a tool for determining the recoil threshold of any beta-emitting radionuclide. PMID- 19297177 TI - A hybrid method to determine efficiency curve of HPGe detectors. AB - A method was developed for determining the efficiency calibration curve of HPGe detectors between 26 and 1770 keV. A simple simulation based on the attenuation of gamma-rays in pure germanium was used together with efficiency calibrations performed for three detector-source distances: 5, 12 and 30 cm. The simple simulation model required an energy dependent term with three parameters and these parameters were determined from the fit of the simulated data to the measured efficiency of the detector. The efficiency response of the detector determined this way was then tested with other certified sources to verify the validity of the method. PMID- 19297178 TI - Classification of materials for explosives from prompt gamma spectra by using principal component analysis. AB - Data from the elemental composition-ratios and experimental prompt gamma spectra of samples were used to develop suitable discriminant classes for suspect samples. The collected data and gamma spectra were applied to principal component analysis (PCA) to discriminate explosives from non-explosive materials. PMID- 19297179 TI - A study on radiation technological degradation of organic chloride wastewater- exemplified by TCE and PCE. AB - This paper describes the potential of using gamma radiation technology to degrade trichloroethylene (TCE) and perchloroethylene (PCE) wastewater. The experimental method is divided into two parts: (1) using the gamma-ray to irradiate the TCE and PCE solution, the dose-rate is 10Gy/minute, the irradiation dosage is 0 2.5kGy and (2) self-making the UV irradiation system, the tube specification is 254nm and 6W, and turning on 8 tubes at the same time to make the irradiation. The efficiency of degradation ratio for gamma-ray is better than UV in the range of 0.1-250ppm; for example, as for the concentration of 0.1ppm, when TCE is degraded to D(90) and T(90), the gamma-ray only needed 46.7Gy and took about 4.67 minutes, but UV needed to take about 28.1 minutes. The dose-concentration equations of TCE and PCE are: TCE: y=44.58+8.832x, R(2)=0.999; and PCE: y=81.33+12.81x, R(2)=0.997. We verified that the radiation technology is able to effectively degrade the organic chlorine wastewater without yielding the secondary pollution, and the TCE and PCE that degraded by using gamma-ray will be reached US-EPA and Taiwan Effluent Standard (5ppb). PMID- 19297180 TI - The measurement of 129I for the cement and the paraffin solidified low and intermediate level wastes (LILWs), spent resin or evaporated bottom from the pressurized water reactor (PWR) nuclear power plants. AB - In this paper a relatively simple and low cost analysis procedure to apply to a routine analysis of (129)I in low and intermediate level radioactive wastes (LILWs), cement and paraffin solidified evaporated bottom and spent resin, which are produced from nuclear power plants (NPPs), pressurized water reactors (PWR), is presented. The (129)I is separated from other nuclides in LILWs using an anion exchange adsorption and solvent extraction by controlling the oxidation and reduction state and is then precipitated as silver iodide for counting the beta activity with a low background gas proportional counter (GPC). The counting efficiency of GPC was varied from 4% to 8% and it was reversely proportional to the weight of AgI by a self absorption of the beta activity. Compared to a higher pH, the chemical recovery of iodide as AgI was lowered at pH 4. It was found that the chemical recovery of iodide for the cement powder showed a lower trend by increasing the cement powder weight, but it was not affected for the paraffin sample. In this experiment, the overall chemical recovery yield of the cement and paraffin solidified LILW samples and the average weight of them were 67+/-3% and 5.43+/-0.53 g, 70+/-7% and 10.40+/-1.60 g, respectively. And the minimum detectable activity (MDA) of (129)I for the cement and paraffin solidified LILW samples was calculated as 0.070 and 0.036 Bq/g, respectively. Among the analyzed cement solidified LILW samples, (129)I activity concentration of four samples was slightly higher than the MDA and their ranges were 0.076-0.114 Bq/g. Also of the analyzed paraffin solidified LILW samples, five samples contained a little higher (129)I activity concentration than the MDA and their ranges were 0.036-0.107 Bq/g. PMID- 19297181 TI - A novel approach to prepare (90)Y-EGMP patches for superficial brachytherapy. AB - A novel method to prepare (90)Y-EGMP patches has been developed for brachytherapy applications.EGMP films of 1cmx1cm size, incorporating approximately 185MBq of (90)Y were prepared and sealed between thin plastic sheets with uniform distribution of (90)Y. The leachability of (90)Y from radioactive patches was less than 0.01%. There was no leakage of radioactivity from radioactive patches, when tested in water or saline. The studies related to establish therapeutic efficacy of these patches are warranted. PMID- 19297182 TI - Investigations of the (nat)Ti(p,x)(43,44m,44g,46,47,48)Sc,(48)V nuclear processes up to 40MeV. AB - Independent and cumulative production cross-sections for the (nat)Ti(p,x)(48)V, (43,44m,44g,46,47,48)Sc nuclear processes are reported here, for the energy region of 4-38MeV by using a stacked-foil activation technique. Measured data were critically compared with the earlier reported values, and also with the theoretical data from the TALYS and ALICE-IPPE codes. The measured (nat)Ti(p,x)(48)V reaction is important for charged particle beam monitoring purposes, whereas the (43,44,47)Sc radionuclide have various practical applications in nuclear medicine. PMID- 19297183 TI - Flow-rate measurements of a dual-phase pipe flow by cross-correlation technique of transmitted radiation signals. AB - The flow rate measurements of dual-phase pipe flow were conducted with a couple of sealed radioisotopes for the cross-correlation technology. The flow was produced in an acrylic tubing of which internal diameter is 8cm and into which pressurized nitrogen gas was periodically injected. Under the given conditions where the distance between two sources is four times of the diameter, N/S ratio ranges from 0.12 to 0.15 and the data sampling time is 4ms, the measured flow rates were estimated with the maximum relative error of 1.7%. From the subsequent experiments, it was proven that the closer the distance between the two sealed sources is, the higher the precision of measurement result is. It is anticipated that the industrial application of the technique for flow rate measurements in an in-service process can play an important role for monitoring multi-phase process media in petrochemical and refinery industries. PMID- 19297184 TI - Environmental stresses induce health-promoting phytochemicals in lettuce. AB - Plants typically respond to environmental stresses by inducing antioxidants as a defense mechanism. As a number of these are also phytochemicals with health promoting qualities in the human diet, we have used mild environmental stresses to enhance the phytochemical content of lettuce, a common leafy vegetable. Five week-old lettuce (Lactuca sativa L.) plants grown in growth chambers were exposed to mild stresses such as heat shock (40 degrees C for 10 min), chilling (4 degrees C for 1d) or high light intensity (800 micromolm(-2)s(-1) for 1d). In response to these stresses, there was a two to threefold increase in the total phenolic content and a significant increase in the antioxidant capacity. The concentrations of two major phenolic compounds in lettuce, chicoric acid and chlorogenic acid, increased significantly in response to all the stresses. Quercetin-3-O-glucoside and luteolin-7-O-glucoside were not detected in the control plants, but showed marked accumulations following the stress treatments. The results suggest that certain phenolic compounds can be induced in lettuce by environmental stresses. Of all the stress treatments, high light produced the greatest accumulation of phenolic compounds, especially following the stress treatments during the recovery. In addition, key genes such as phenylalanine ammonia-lyase (PAL), l-galactose dehydrogenase (l-GalDH), and gamma-tocopherol methyltransferase (gamma-TMT) involved in the biosynthesis of phenolic compounds, ascorbic acid, and alpha-tocopherol, respectively, were rapidly activated by chilling stress while heat shock and high light did not appear to have an effect on the expression of PAL and gamma-TMT. However, l-GalDH was consistently activated in response to all the stresses. The results also show that these mild environmental stresses had no adverse effects on the overall growth of lettuce, suggesting that it is possible to use mild environmental stresses to successfully improve the phytochemical content and hence the health-promoting quality of lettuce with little or no adverse effect on its growth or yield. PMID- 19297185 TI - Guideline for resuscitation in cardiac arrest after cardiac surgery. AB - The Clinical Guidelines Committee of the European Association for Cardio-Thoracic Surgery provides this professional view on resuscitation in cardiac arrest after cardiac surgery. This document was created using a multimodal methodology for evidence generation including the extrapolation of existing guidelines from the International Liaison Committee on Resuscitation where possible, our own structured literature reviews on issues particular to cardiac surgery, an international survey on resuscitation hosted by CTSNet and manikin simulations of potential protocols. This protocol differs from existing generic guidelines in a number of areas, the most import of which are the following: successful treatment of cardiac arrest after cardiac surgery is a multi-practitioner activity with six key roles that should be allocated and rehearsed on a regular basis; in ventricular fibrillation, three sequential attempts at defibrillation (where immediately available) should precede external cardiac massage; in asystole or extreme bradycardia, pacing (where immediately available) should precede external cardiac massage; where the above measures fail, and in pulseless electrical activity, early resternotomy is advocated; adrenaline should not be routinely given; protocols for excluding reversible airway and breathing complications and for safe emergency resternotomy are given. This guideline is subject to continuous informal review, and when new evidence becomes available. PMID- 19297186 TI - Inhibitory effect of leptin on growth hormone secretion of GH3 cells: involvement of cell proliferation, apoptosis and intracellular free Ca2+. AB - Recent studies have identified leptin and leptin receptors in the pituitary of different species, which suggest that there may be endocrine and paracrine regulatory roles between leptin-producing cells and cells with leptin receptor in pituitary, including growth and secretion of GH cells. The aim of this study was to investigate the effects of leptin on growth hormone (GH) secretion of GH3 cell. GH3 cells were cultured and treated with leptin. Cell proliferation was evaluated by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay, distribution of cell cycle and rate of apoptosis determined by flow cytometry and fluorescence microscopy, and intracellular free Ca(2+) levels ([Ca(2+)](i)) of single GH3 cells measured by a laser scanning confocal microscope. Leptin (10(-9)-10(-7)mol/L) at 1day or longer of treatment inhibited the basal growth hormone secretion of GH3 cells (P<0.05), but had no significant effect on short-term treatment. Leptin inhibited cell proliferation, reduced the proportion of cells in DNA synthesis period (S phase) to inhibit DNA synthesis of GH3 cells, and accelerated cell apoptosis of GH3 cells. Furthermore, the level of [Ca(2+)](i) of single GH3 cell was found to decrease immediately upon the addition of leptin (10(-8)mol/L). Leptin inhibits the basal GH secretion of GH3 cells, which may be due to the inhibition of proliferation, DNA synthesis and advanced apoptosis of GH3 cell. The inhibition of leptin on GH synthesis and secretion may be related to intracellular free Ca(2+) level. PMID- 19297187 TI - SALDI-MS signal enhancement using oxidized graphitized carbon black nanoparticles. AB - The signal intensity of low-molecular-weight compounds analyzed using surface assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-TOF MS) was significantly enhanced when oxidized graphitized carbon black (GCB) particles were used as the desorption/ionization surface. The surface of oxidized GCB contains more carboxylic acid groups than non-oxidized GCB. Carboxylic acid groups enhance the efficiency of the ionization process and the desorption of more hydrophobic compounds. A common pharmaceutical compound, propranolol, was successfully extracted from Baltic Sea blue mussels and quantified using oxidized GCB as the SALDI surface, whereas deuterated propranolol was used as the internal standard. The calibration curve showed a wide linear dynamic range of response (0.1-20 microg/mL) and good reproducibility (RSD < 10%). It was not possible to detect propranolol in Baltic Sea blue mussels when non-oxidized GCB was used as the SALDI surface. PMID- 19297188 TI - Evaluation of effects of bivalent cations on the formation of purine-rich triple helix DNA by ESI-FT-MS. AB - The GGA triplet repeats are widely dispersed throughout eukaryotic genomes. (GGA)n or (GGT)n oligonucleotides can interact with double-stranded DNA containing (GGA:CCT)n to form triple-stranded DNA. The effects of 8 divalent metal ions (3 alkaline-earth metals and 5 transition metals) on formation of these purine-rich triple-helix DNA were investigated by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FT-MS). In the absence of metal ions, no triplex but single-strand, duplex, and purine homodimer ions were observed in mass spectra. The triple-helix DNA complexes were observed only in the presence of certain divalent ions. The effects of different divalent cations on the formation of purine-rich triplexes were compared. Transition-metal ions, especially Co(2+) and Ni(2+), significantly boost the formation of triple helix DNA, whereas alkaline-earth metal ions have no positive effects on triplex formation. In addition, Ba(2+) is notably beneficial to the formation of homodimer instead of triplex. PMID- 19297189 TI - The use of ESI-MS to probe the binding of divalent cations to calmodulin. AB - Proteins have evolved with distinct sites for binding particular metal ions. This allows metalloproteins to perform a myriad of specialized tasks with conformations tailor-made by the combination of its primary sequence and the effect on this of the ligated metal ion. Here we investigate the selectivity of the calcium trigger protein calmodulin for divalent metal ions. This ubiquitous and highly abundant protein exists in equilibrium between its apo and its holo form wherein four calcium ions are bound. Amongst its many functions, calmodulin modulates the calcium concentration present in cells, but this functional property renders it a target for competition from other metal ions. We study the competition posed by four other divalent cations for the calcium binding sites in calmodulin using electrospray ionization mass spectrometry (ESI-MS). We have chosen two other group II cations Mg(2+), Sr(2+), and two heavy metals Cd(2+), Pb(2+). The ease with which each of these metals binds to apo and to holo CaM[4Ca] is described. We find that each metal ion has different properties with respect to calmodulin binding and competition with calcium. The order of affinity for apo CaM is Ca(2+) >> Sr(2+) approximately Mg(2+) > Pb(2+) approximately Cd(2+). In the presence of calcium the affinity alters to Pb(2+) > Ca(2+) > Cd(2+) > Sr(2+) > Mg(2+). Once complexes have been formed between the metal ions and protein (CaM:[xM]) we investigate whether the structural change which must accompanies calcium ligation to allow target binding takes place for a given CaM:[xM] system. We use a 20 residue target peptide, which forms the CaM binding site within the enzyme neuronal nitric-oxide synthase. Our earlier work (Shirran et al. 2005) [1] has demonstrated the particular selectivity of this system for CaM:4Ca(2+). We find that along with Ca(2+) only Pb(2+) forms complexes of the form CaM:4M(2+):nNOS. This work demonstrates the affinity for calcium above all other metals, but also warns about the ability of lead to replace calcium with apparent ease. PMID- 19297191 TI - Focus in honor of Dr. Julia Laskin, recipient of the 2008 Biemann Medal. Interview by Richard A. J. O'Hair and K. W. Michael Siu. PMID- 19297190 TI - Periodic sequence distribution of product ion abundances in electron capture dissociation of amphipathic peptides and proteins. AB - The rules for product ion formation in electron capture dissociation (ECD) mass spectrometry of peptides and proteins remain unclear. Random backbone cleavage probability and the nonspecific nature of ECD toward amino acid sequence have been reported, contrary to preferential channels of fragmentation in slow heating based tandem mass spectrometry. Here we demonstrate that for amphipathic peptides and proteins, modulation of ECD product ion abundance (PIA) along the sequence is pronounced. Moreover, because of the specific primary (and presumably secondary) structure of amphipathic peptides, PIA in ECD demonstrates a clear and reproducible periodic sequence distribution. On the one hand, the period of ECD PIA corresponds to periodic distribution of spatially separated hydrophobic and hydrophilic domains within the peptide primary sequence. On the other hand, the same period correlates with secondary structure units, such as alpha-helical turns, known for solution-phase structure. Based on a number of examples, we formulate a set of characteristic features for ECD of amphipathic peptides and proteins: (1) periodic distribution of PIA is observed and is reproducible in a wide range of ECD parameters and on different experimental platforms; (2) local maxima of PIA are not necessarily located near the charged site; (3) ion activation before ECD not only extends product ion sequence coverage but also preserves ion yield modulation; (4) the most efficient cleavage (e.g. global maximum of ECD PIA distribution) can be remote from the charged site; (5) the number and location of PIA maxima correlate with amino acid hydrophobicity maxima generally to within a single amino acid displacement; and (6) preferential cleavage sites follow a selected hydrogen spine in an alpha-helical peptide segment. Presently proposed novel insights into ECD behavior are important for advancing understanding of the ECD mechanism, particularly the role of peptide sequence on PIA. An improved ECD model could facilitate protein sequencing and improve identification of unknown proteins in proteomics technologies. In structural biology, the periodic/preferential product ion yield in ECD of alpha helical structures potentially opens the way toward de novo site-specific secondary structure determination of peptides and proteins in the gas phase and its correlation with solution-phase structure. PMID- 19297192 TI - Chiral differentiation of some cyclopentane and cyclohexane beta-amino acid enantiomers through ion/molecule reactions. AB - Chiral differentiation of four enantiomeric pairs of beta-amino acids, cis (1R,2S)-, cis-(1S,2R)-, trans-(1R,2R)-, and trans-(1S,2S)-2 aminocyclopentanecarboxylic acids (cyclopentane beta-amino acids), and cis (1R,2S)-, cis-(1S,2R)-, trans-(1R,2R)-, and trans-(1S,2S)-2 aminocyclohexanecarboxylic acids (cyclohexane beta-amino acids) was performed successfully by using host-guest complexes and ion/molecule reactions. The experiments were conducted by using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. The effect of a chiral host molecule was tested by using three different host compounds; (+)-(18-Crown-6)-2,3,11,12 tetracarboxylic acid, (-)-(18-Crown-6)-2,3,11,12-tetracarboxylic acid, and beta cyclodextrin. This is the first time that small enantiomeric pairs with two chiral centers have been differentiated using ion/molecule reactions and host guest complexes. PMID- 19297193 TI - Isomer separation and gas-phase configurations of organoruthenium anticancer complexes: ion mobility mass spectrometry and modeling. AB - We have used ion mobility-mass spectrometry combined with molecular modeling for the separation and configurational analysis of three low-molecular-weight isomeric organoruthenium anticancer complexes containing ortho-, meta-, or para terphenyl arene ligands. The isomers were separated using ion mobility based on traveling-wave technology and the experimentally determined collision cross sections were compared to theoretical calculations. Excellent agreement was observed between the experimentally and theoretically derived measurements. PMID- 19297194 TI - FluBlok, a next generation influenza vaccine manufactured in insect cells. AB - FluBlok, a recombinant trivalent hemagglutinin (rHA) vaccine produced in insect cell culture using the baculovirus expression system, provides an attractive alternative to the current egg-based trivalent inactivated influenza vaccine (TIV). Its manufacturing process presents the possibility for safe and expeditious vaccine production. FluBlok contains three times more HA than TIV and does not contain egg-protein or preservatives. The high purity of the antigen enables administration at higher doses without a significant increase in side effects in human subjects. The insect cell-baculovirus production technology is particularly suitable for influenza where annual adjustment of the vaccine is required. The baculovirus-insect expression system is generally considered a safe production system, with limited growth potential for adventitious agents. Still regulators question and challenge the safety of this novel cell substrate as FluBlok continues to advance toward product approval. This review provides an overview of cell substrate characterization for expresSF cell line used for the manufacturing of FluBlok. In addition, this review includes an update on the clinical development of FluBlok. The highly purified protein vaccine, administered at three times higher antigen content than TIV, is well tolerated and results in stronger immunogenicity, a long lasting immune response and provides cross-protection against drift influenza viruses. PMID- 19297195 TI - Fatal necrotic pancreatitis as a rare complication after thoracoabdominal aortic surgery. PMID- 19297196 TI - Esmolol reduces perioperative ischemia in cardiac surgery: a meta-analysis of randomized controlled studies. AB - OBJECTIVE: beta-Blockers were associated with a reduction of mortality and morbidity in noncardiac surgery until recently when the POISE trial showed that beta-blockers could be harmful in the perioperative period because of hypotension and bradycardia. Esmolol is an ultra-short-acting beta-blocker mostly used in emergency and high-risk patients. The authors performed a meta-analysis to evaluate the clinical effects of esmolol in cardiac surgery. DESIGN: Meta analysis. SETTING: Hospitals. PARTICIPANTS: A total of 778 patients from 20 randomized trials. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULT: Three investigators independently searched BioMedCentral and PubMed. Inclusion criteria were random allocation to treatment and comparison of esmolol versus other drugs, placebo, or standard of care in cardiac surgery. Exclusion criteria were duplicate publications, nonhuman experimental studies, and no data on clinical outcomes. The use of esmolol was associated with a significant reduction of myocardial ischemia episodes (15/122 [12.2%] in the esmolol group v 36/140 [25.7%] in the control arm, odds ratio [OR] =0.42 [0.23-0.79], p = 0.007) and development of arrhythmias after cardiopulmonary bypass (15/65 [23.07%] v 23/64 [35.9%], OR = 0.42 [0.18-1.01], p = 0.05). The authors did not find a reduction in the use of inotropic drugs in esmolol-treated patients (29/153 [18.9%] v 48/146 [32.8%], OR = 0.43 [0.16-1.10], p = 0.08). Esmolol-treated patients had more episodes of bradycardia (19/129 [14.72%] v 3/133 [2.25%], OR = 5.49 [2.21 13.62], p = 0.0002) and hypotension (28/113 [24.77%] v 14/119 [11.76%], OR = 2.73 [0.83-9.04], p = 0.10). CONCLUSIONS: Esmolol reduces the incidence of myocardial ischemia and arrhythmias in cardiac surgery. An increase in bradycardia was noted as well. PMID- 19297197 TI - When 9 is not on the right: implications from number-form synesthesia. AB - Number-form synesthetes consciously experience numbers in spatially-defined locations. For non-synesthete individuals, a similar association of numbers and space appears in the form of an implicit mental number line as signified by the distance effect-reaction time decreases as the numerical distance between compared numbers increases. In the current experiment, three number-form synesthetes and two different non-synesthete control groups (Hebrew speaking and English speaking) performed a number comparison task. Synesthete participants exhibited a sizeable distance effect only when presented numbers were congruent with their number-form. In contrast, the controls exhibited a distance effect regardless of congruency or presentation type. The findings suggest that: (a) number-form synesthesia impairs the ability to represent numbers in a flexible manner according to task demands; (b) number-form synesthesia is a genuine tangible experience, triggered involuntarily; and (c) the classic mental number line can be more pliable than previously thought and appears to be independent of cultural-lingo direction. PMID- 19297198 TI - Rotator cuff repair fluid extravasation characteristics are influenced by repair technique. AB - BACKGROUND: This study evaluates rotator cuff repair fluid extravasation characteristics for different rotator cuff repair techniques. METHODS: Eight fresh-frozen cadaveric shoulders were dissected free of soft tissues, with the glenohumeral joint capsule and rotator cuff muscles being left intact. A custom fluid infusion device was used to deliver fluid at constant pressure into the glenohumeral joint. The shoulders were tested in conditions of (1) intact rotator cuff, (2) supraspinatus tear, (3) repaired supraspinatus tear with a single-row technique, and (4) repaired supraspinatus tear with a double-row suture bridge technique. RESULTS: The volume per minute of saline solution extravasation for single-row repair and double-row suture bridge repair was 48.53 mL/min and 11.73 mL/min, respectively, at 2 psi; 73.3 _ 24.1 mL/min and 24.5 _ 19.7 mL/min, respectively, at 3 psi; and 95.2 _ 22.6 mL/min and 39.2 _ 23.8 mL/min, respectively, at 4 psi. There was a statistically significant greater fluid extravasation for the single-row repair compared with the double-row suture bridge repair at all 3 pressures tested (P < .05). CONCLUSION: Single-row rotator cuff repair exposes the healing zone to greater extravasation of fluid compared with double-row suture bridge repair. Therefore, double-row repair potentially enhances rotator cuff healing. LEVEL OF EVIDENCE: Controlled laboratory study. PMID- 19297199 TI - Survival of the glenoid component in shoulder arthroplasty. AB - INTRODUCTION: This study was conducted to determine the survival of different glenoid component designs, assess the reasons for revision surgery, and identify patient and diagnostic factors that influence this need. METHODS: Between January 1, 1984, and December 31, 2004, 1337 patients underwent 1542 total shoulder arthroplasties with 6 types of glenoid components: Neer II all-polyethylene, Neer II metal-backed, Cofield 1 metal-backed bone-ingrowth, Cofield 1 all-poly keeled, Cofield 2 all-poly keeled, and Cofield 2 all-poly pegged. RESULTS: Revision was required in 125 shoulders for glenoid component failure. Survival rates free of revision by glenoid implant type at 5, 10, and 15 years were, respectively, 96%, 96%, and 95% for 99 Neer II all-poly; 96%, 94%, and 89% for 316 Neer II metal backed; 86%, 79%, and 67% for 316 Cofield 1 metal-backed; 94%, 94%, and 87% for 18 Cofield 1 all-poly; 99%, 94%, and 89% for 497 Cofield 2 all-poly keeled; and 99% at 5 years for 358 Cofield 2 all-poly pegged. Glenoid component type was significantly associated with component revision (P < .001). Male gender was associated with a higher risk of revision (P < .001). Compared with degenerative arthritis, posttraumatic arthritis (P = .02) and avascular necrosis (P = .06) were associated with increased risk of revision. CONCLUSIONS: Survival is improved with cemented all-polyethylene glenoid components. Revision of cemented all-polyethylene components may be lessened with the use of pegged components in early follow-up. Male gender and the operative diagnoses of posttraumatic arthritis or avascular necrosis are associated with an increased risk of failure. LEVEL OF EVIDENCE: Level IV, Case Series, Treatment Study. PMID- 19297201 TI - Four-segment classification of proximal humeral fractures revisited: a multicenter study on 509 cases. AB - BACKGROUND: The 4-segment classification of Neer (1970) was revised in 2002 with the inclusion of valgus-impacted fractures. However, the range of possible fracture types covered by this classification is not clear. With the aim to clarify actual fracture patterns of the proximal humerus, a retrospective, multicenter study was conducted. METHODS: We retrospectively reviewed 509 cases for which anteroposterior and trans-scapular lateral views had been taken at the time of injury. Initially, three examiners from three facilities, independently read the radiographs of each case. In the second and third phases, for more difficult cases, consensus opinions were reached. RESULTS: We found that 501 (98%) of the 509 fractures had an appropriate category in the revised Neer classification. There were 185 cases (36%) of one-part fractures, followed by 156 cases (31%) of 2-part surgical neck fracture, 60 cases (12%) of 2-part greater tuberosity fracture (dislocation included), 45 cases (8.8%) of 3-part fracture involving the greater tuberosity and the surgical neck (dislocation included), 31 cases (6.1%) of 4-part fracture (dislocation included), and 17 cases (3.3%) of valgus-impacted fracture.Eight fractures (1.6%) could not be classified with the revised Neer classification. They were "3-part" fractures in terms of the number of displaced segments but had a complete anatomic neck fracture. CONCLUSION: We conclude that it would be appropriate to use the revised Neer classification in clinical practice. We would emphasize that, when there are 3 displaced segments, close attention should be paid to the fracture line on the anatomic neck. LEVEL OF EVIDENCE: Level 4; Diagnostic study, case series. PMID- 19297202 TI - Psychometric properties of the shortened disabilities of the Arm, Shoulder, and Hand Questionnaire (QuickDASH) and Numeric Pain Rating Scale in patients with shoulder pain. AB - PURPOSE: To examine the psychometric properties of the Shortened Disabilities of the Arm, Shoulder, and Hand Questionnaire (QuickDASH) and the Numeric Pain Rating Scale (NPRS) in patients with shoulder pain. METHODS: Single-group repeated measures design in which 101 patients presenting to physical therapy completed the QuickDASH and the NPRS at the baseline examination and at a follow-up visit. At the follow-up all patients also completed the Global Rating of Change (GRC), which was used to dichotomize patients as improved or stable. Baseline and follow up scores were used to determine the test-retest reliability, construct validity and minimal levels of detectable and clinically important change for both the QuickDASH and NPRS. RESULTS: Test-retest reliability was 0.90 for the QuickDASH and 0.74 for the NPRS. The minimal clinically important difference (MCID) was 8.0 points for the QuickDASH and 1.1 for the NPRS. CONCLUSION: The NPRS and QuickDASH exhibit good test-retest reliability and responsiveness in patients with shoulder pain. PMID- 19297203 TI - Geometric variables in anatomic replacement of the proximal humerus: how much prosthetic geometry is necessary? AB - BACKGROUND: Contemporary prosthetic designs for replacement of the proximal humerus offer a range of variable geometry aiming for restoration of the anatomy. This study evaluated five fictitious prosthetic scenarios looking to answer: How much prosthetic geometry is needed to achieve a minimum standard of anatomic reconstruction? METHODS: Five prosthetic scenarios graduated in complexity in the number of inclination angles offered, number of head heights available and the number of offset positions of the head were compared to a database of known proximal humeral anatomy. Inclination ranged from a single inclination of 42 degrees to six inclination angles, head height ranged from a single head thickness to five possible head heights at any of four radii of curvature, and offset ranged from one to four variable positions that allowed for an increasing number of head positions relative to the stem. The five scenarios were compared to the database of normal anatomy using a computer optimization algorithm that aimed to minimize the displacement of the center of rotation and articular surface. For the purposes of this analysis, the minimum standard of reconstruction was defined as one that re-positioned the center of rotation within 4mm maintaining articular surface arc within 30 degrees for 95% of all specimens. RESULTS: The mean displacement of the center of rotation and articular surface decreased with progressive increase in the geometric complexity of the prosthetic scenarios. However, the best fit observed in the entire analysis occurred in the simplest scenario with a single inclination angle (42 degrees) because the available prosthetic geometry happened to match one of the specimens. The worst fit also occurred in this scenario despite the specimen having a similar inclination angle (39 degrees) to that of the scenario due to differential offsets, which were more challenging for the algorithm than inclination. The most challenging anatomy for the computer algorithm to match was that with extreme offset of the head. The threshold 95% C.I. that was the stated goal of this study was achieved with the geometric complexity provided in the scenario with two inclination angles, three head heights (12 head sizes) and two offset positions of the taper mechanism. CONCLUSION: In this analysis, even the simplest prosthetic scenario allowed for replication of normal anatomy within a range comparable to some existing third generation prosthetic systems. From a geometric perspective, surgeons should be able to re-approximate normal anatomy with a variety of implant configurations. PMID- 19297204 TI - Tetracycline labeling as a measure of humeral head viability after 3- or 4-part proximal humerus fracture. AB - HYPOTHESIS: Specifically located humeral head biopsies from three-part proximal humerus fractures taken at the time of hemiarthroplasty will show greater tetracycline labeling under fluorescent microscopy than those biopsies taken from four-part fractures. Additionally, biopsies from younger patients would show greater fluorescence than older patients. MATERIAL AND METHODS: Nineteen consecutive adult patients (range 43-83 years) underwent hemiarthroplasty as definitive treatment for 20 displaced three- and four-part proximal humerus fractures after having received 500 mg of tetracycline hydrochloride orally every six hours for the immediate five preoperative days. Humeral head biopsies were taken from four pre-determined locations intraoperatively. The biopsies were prepared and analyzed with fluorescent microscopy. RESULTS: All specimens in each biopsy location demonstrated fluorescence. There was no difference between the mean rank gray values for the four biopsy locations (p = 0.78 with the Friedman test). There was no difference between the mean rank gray values for the four biopsy locations when analyzed according to three-part vs four-part fracture (p > 0.05 with the Mann-Whitney test). There was an inverse relationship between age and fluorescence for the anterosuperior biopsy location (p = 0.033 with Spearman correlation). DISCUSSION: Vascular supply is preserved in displaced three- and four-part proximal humerus fractures. With intact vascularity to the humeral head, head-preserving techniques utilizing stable, site-specific fixation and minimal dissection should be considered in the treatment of displaced three- and four-part proximal humerus fractures. CONCLUSION: Vascular supply is preserved in displaced three- and four-part proximal humerus fractures, especially in younger patients in the anterosuperior aspect of the humeral head. LEVEL OF EVIDENCE: Basic Science Study. PMID- 19297205 TI - How to quote Gustilo open fracture classification? PMID- 19297206 TI - Cortical excitability in juvenile myoclonic epileptic patients and their asymptomatic siblings: a transcranial magnetic stimulation study. AB - In this study, we aimed to evaluate motor cortical excitability changes in patients with juvenile myoclonic epilepsy (JME) and their asymptomatic siblings (AS) using single-pulse transcranial magnetic stimulation (spTMS). 21 patients with JME and their 21 AS were compared to 20 healthy controls. All of JME patients were receiving antiepileptic therapy and their seizures were well controlled. Firstly, standard EEG examinations and then TMS studies were performed. Resting motor threshold (RMT), motor evoked potential (MEP) amplitudes, the durations of central motor conduction time (CMCT) and cortical silent period (CSP) were measured. After TMS studies, EEG recordings were repeated in an hour to evaluate any effect of TMS study on EEG. There were no significant differences between the first and second EEG recordings. No seizures were recorded during and after the TMS study. RMT was found higher in JME patients than AS and normal controls. There were no significant differences between cortical MEP amplitudes and MEP amplitude/CMAP (compound muscle action potential) amplitude ratio in all three groups. CMCT duration was shorter in JME patients than AS. CSP durations of JME patients were found to be longer than controls. In AS, CSP durations were also found to be longer than controls but this difference was not found statistically significant. Our results suggested that although high MT may be related to antiepileptic therapy, the prolongation of CSP duration may reflect impairment of supraspinal and/or intracortical inhibitory mechanism in JME. To eliminate the drug effect, further studies are needed in newly diagnosed JME patients without medication and large series of their asymptomatic siblings. PMID- 19297207 TI - Whole-body MR angiography with body coil acquisition at 3 T in patients with peripheral arterial disease using the contrast agent gadofosveset trisodium. AB - RATIONALE AND OBJECTIVES: Whole-body magnetic resonance angiography (WB-MRA) at 3 T with body coil acquisition has not previously been investigated. In this study, WB-MRA was performed in this manner using the blood pool contrast agent gadofosveset trisodium. MATERIALS AND METHODS: Eleven consecutive patients (five men, six women) with symptomatic peripheral arterial disease (two with critical limb ischemia, nine with claudication) were examined. Conventional digital subtraction angiography (DSA) of the aorta and the inflow and runoff arteries was used as the reference method. WB-MRA was performed using four slightly overlapping stations covering the arteries from the neck to the ankles. The arterial system was divided into 42 segments that were analyzed for the presence of significant arterial disease (> or =50% luminal narrowing or occlusion) by two blinded observers. RESULTS: Sensitivities for detecting a significant arterial lesion with WB-MRA using gadofosveset as the contrast agent were 0.66 (95% confidence interval [CI], 0.49-0.79) and 0.68 (95% CI, 0.52-0.81) for the two observers. Specificities were 0.82 (95% CI, 0.74-0.88) and 0.93 (95% CI, 0.87 0.96), respectively. Intermodality agreement between WB-MRA and DSA was moderate to good, with overall kappa values of 0.44 (95% CI, 0.29-0.59) and 0.63 (95% CI, 0.5-0.77) for the two observers. Interobserver agreement for WB-MRA was good, at kappa = 0.60 (95% CI, 0.50-0.71). CONCLUSION: WB-MRA at 3 T with body coil acquisition in patients with peripheral arterial disease showed good reproducibility but only moderate to good agreement with DSA. Further assessment of the method's clinical application is warranted. PMID- 19297208 TI - Early radiation-induced bone marrow injury: serial MR imaging during initial 4 weeks after irradiation. AB - RATIONALE AND OBJECTIVES: Magnetic resonance (MR) imaging has been widely used to detect bone marrow (BM) changes after radiotherapy. However, little information about the dynamic MR appearance of early radiation-induced BM injury is available. This experimental study was designed to determine the MR appearance of irradiated BM during the initial 4 weeks after irradiation. MATERIALS AND METHODS: After focal BM irradiation (20 Gy, single dose, x-ray), 12 of 20 rabbits underwent serial MR studies weekly from days 7 to 28; eight rabbits were used for histologic investigation on days 7, 14, 21, and 28 after irradiation. RESULTS: Under microscopy, early BM changes after irradiation consisted of sinusoid dilatation and congestion, followed by a progressive decrease in cellularity and later fat degeneration. All irradiated BM showed relative hyperintensity on short inversion time inversion recovery (STIR) imaging from days 7 to 21 after irradiation and increased enhancement with gadolinium diethylenetriamine pentaacetic acid (DTPA) administration from days 7 to 28 after irradiation. However, on STIR imaging and gadolinium DTPA enhancement, the relative signal intensity of irradiated BM appeared to decline in a time-dependent way. On fast spin-echo (FSE) T1-weighted imaging, relative hyperintensity was detected in irradiated BM from day 21 after irradiation. On fat-suppressed FSE T1-weighted imaging, a slight increase in signal intensity was shown in some irradiated BM (in five of 12 rabbits) on day 7 after irradiation. CONCLUSION: STIR imaging was sensitive to early BM congestion and sinusoidal dilatation, spin-echo T1-weighted imaging was effective in detecting later fatty degeneration in irradiated BM, and gadolinium DTPA enhancement may contribute to the evaluation of BM vascular injury in response to irradiation. PMID- 19297209 TI - In vivo assessment of mouse hindleg intramyocellular lipids by 1H-MR spectroscopy. AB - RATIONALE AND OBJECTIVES: (1)H-magnetic resonance spectroscopy ((1)H-MRS) has proved to be the sole in vivo technique able to measure intramyocellular lipids (IMCL) in both humans and animals. Mouse models are now widely used for physiologic studies and drug discovery. However, IMCL assessment using (1)H-MRS is hindered in this animal model by the small muscle size and strong contamination from the extramyocellular lipid (EMCL) signal. The objective of this study was to the use of (1)H-MRS for IMCL quantification in mice at different ages. MATERIALS AND METHODS: Noninvasive IMCL quantification was performed at 7 T in tibialis anterior (TA) muscles of healthy male C57/BL6 mice (n = 9; age, 13.6 +/- 1 months), db/db mice (n = 4), and their C57BL/KSJ control littermates (n = 4) at 7 and 17 weeks of age. RESULTS: The IMCL content of diabetic mice TA was significantly higher than their littermates (2.41 +/- 0.5 vs. 1.21 +/- 0.35, P < .01). An age effect was observed, with TA IMCL levels being lower in older than younger control mice, but increasing between 7 and 17 weeks in the db/db mice. CONCLUSIONS: The feasibility of (1)H-MRS spectroscopy was demonstrated in mice muscle, despite its small size, and used to assess IMCL content in db/db mice. PMID- 19297210 TI - Thoracodorsal artery as a collateral source to the artery of Adamkiewicz after endovascular aneurysm repair for descending thoracic aortic aneurysm. AB - Paraplegia is one of the most tragic complications following endovascular aneurysm repair (EVAR) for descending thoracic aortic aneurysm (DTAA). Collateral circulation to the artery of Adamkiewicz (AA) is important to avoid spinal cord ischaemia. We report a case in which the thoracodorsal artery had become a collateral source to the AA. A 71-year-old man had undergone EVAR for DTAA. Three years after EVAR, an angiography demonstrated that the thoracodorsal artery had joined the 11th intercostal artery and had become a collateral source to the AA. The collateral circulation of thoracic wall arteries may play an important role in the postoperative spinal perfusion. PMID- 19297212 TI - Combined aspirin and cilostazol treatment is associated with reduced platelet aggregation and prevention of exercise-induced platelet activation. AB - BACKGROUND: Cilostazol has proven efficacy in increasing walking distance in claudicants, but it has not been demonstrated to be more effective than placebo in secondary cardiovascular prevention. The direct effect of exercise on platelet function remains less well defined. We have investigated the effect of combination treatment with aspirin and cilostazol on platelet activity in claudicants subjected to repeated treadmill exercise. METHODS: Nineteen claudicants completed a double-blind, randomised, controlled, cross-over trial. Each subject received a 2-week course of aspirin (75mg) and placebo and aspirin and cilostazol (100mg twice daily). Following each 2-week treatment period, patients participated in a standardised treadmill test (3.2kmh(-1), 10 degrees incline) walking to maximal claudication distance. The exercise was repeated thrice in total, and blood was sampled before and after exercise. Platelet activation was measured using free platelet counting aggregation, flow cytometry for surface markers of platelet activation and soluble P-selectin assay. RESULTS: Compared to aspirin and placebo, combination treatment with aspirin and cilostazol was associated with reduced arachidonic-acid-induced platelet aggregation (p<0.01, Wilcoxon signed-rank test). Aspirin and placebo treatment were associated with elevated P-selectin expression, platelet-monocyte aggregation and reduced CD42b expression (p<0.05, Wilcoxon signed-rank test) post exercise. No difference was seen in spontaneous platelet aggregation whilst soluble P-selectin was reduced post-exercise with combination treatment with aspirin and cilostazol (p<0.05, Wilcoxon signed-rank test). CONCLUSIONS: Combination treatment with aspirin and cilostazol results in suppression of platelet activation and reduces the effect of exercise on platelets. The benefit seen may be a result of cilostazol enhancing the inhibitory effect of aspirin on the cyclo-oxygenase pathway. PMID- 19297213 TI - A new modular stent graft to reconstruct aortic arch. AB - BACKGROUND: Endovascular repair by stent graft has been developed as a safe and less-invasive treatment for descending thoracic and abdominal aortic diseases. In case of involvement of the aortic arch, the challenge in endovascular repair is to maintain blood flow to the brain and upper extremities. Several studies have been done trying to repair this difficult part of the aorta with different stent grafts, and we have developed a new stent-graft device for aortic arch reconstitution. We implanted the new device in canine models to test its feasibility. METHODS AND RESULTS: The new stent graft was composed of three components: parts I and II were both bifurcated, one with long, narrow limb and the other short and wide, and part III was a tubular component. Ten adult hybrid dogs were operated with the new stent-graft procedure, and eight were successfully implanted with the stent grafts. The technical success rate was 80% (8 of 10). Five dogs survived for 3 months without obvious cerebral, visceral or limb ischaemia. Autopsies showed that the implanted stent grafts were patent and the vital side branches of aortic arch were well preserved. CONCLUSIONS: Our study demonstrates that it is possible to reconstruct aortic arch with the new branched stent grafts. The advantage of this device is that it is modular, more adaptable and surgical bypass could be possibly avoided. PMID- 19297214 TI - New insight in aetiopathogenesis of aortic diseases. AB - BACKGROUND: Knowledge in the aetiopathogeny of aortic disease helps to characterise aortic lesions better and determine the risk of evolution and therapeutic strategies as well. This article focusses on aneurysms and dissections, and excludes causes related to infection, systemic inflammatory diseases and trauma. METHODS AND RESULTS: The biomedical literature of the past 10 years has been reviewed here. Aortic diseases are heterogeneous along the aorta as far as their genetic determinants, contribution of smooth muscle cells, inflammation and thrombus formation are concerned. Degradation of extracellular matrix by proteases causing aortic disease is a 'terminal' event, modulated by genetic background, haemodynamic strain, cellular events and thrombus formation. New genetic determinants of aortic disease have been identified. Proteases degrading the aortic wall are derived from a variety of cell types in addition to macrophages, including neutrophils on the luminal thrombus, mesenchymal and endothelial cells in the wall. Smooth muscle cells contribute to aortic wall homeostasis against inflammation and proteolysis. The degradation of the wall is followed by, or paralleled with, a failure of aortic reconstruction. CONCLUSIONS: Aortic diseases are diverse, and involve a multiplicity of biological systems in the vascular wall and at the interface with blood. Future research needs to unravel distinct cellular and molecular mechanisms causing the clinical events, in particular, dissection, expansion of already formed aneurysms and rupture. PMID- 19297215 TI - Early endothelial dysfunction in young type 1 diabetics. AB - OBJECTIVES: Endothelial dysfunction is a known precursor of atherosclerosis and can be assessed by measuring the brachial artery flow-mediated dilatation (FMD) via ultrasonography. This study investigated endothelial function in young type 1 diabetics without cardiovascular morbidity or diabetes-related pathology. METHODS: Young diabetics and healthy controls were recruited, both meeting strict inclusion and exclusion criteria. To prove absence of subclinical atherosclerosis, intima-media thickness (IMT) measurements at the carotid bifurcation were done in all of them. FMD was measured at the brachial artery. The results were compared using the t-test and the influences of different variables on FMD were assessed using multiple linear regression. RESULTS: Twenty six diabetics (23.4+/-5.8 years) and 36 healthy volunteers (23.1+/-2.8 years) were recruited. The duration of diabetes was 9.2+/-5.3 years; metabolic control was moderate (HbA1c 7.6+/-1.0%) and IMT was normal in both groups. FMD was significantly impaired in type 1 diabetics (7.13+/-0.43 vs. 8.77+/-0.43%; p=0.002). The FMD grade was associated with diabetes and age. Patients with a good metabolic control (HbA1c/=70%) were included in the study. Restenosis was symptomatic in 23 patients and asymptomatic in 29 patients. METHODS: Recurrent carotid artery stenosis was classified based on presentation, and as early intermediate (6 months to 3 years) or late (>3 years). A detailed clinical history was taken and two blood samples were drawn from each patient to determine plasma levels of MMPs and TIMPs along with other biological parameters. Recurrent stenosis was confirmed with computed tomographic angiography. RESULTS: Patients with symptomatic restenosis had significantly (p<0.001) higher active MMP-2 and 9 plasma values and significantly (p<0.001) lower TIMP-1 and -2 plasma values when compared to patients with asymptomatic restenosis. Plasma concentrations of active MMPs were higher and TIMPs lower in patients affected with late recurrent stenosis as compared to early-intermediate restenosis (p<0.001). No differences were recorded in latent MMP plasma values. Multivariate analysis showed that active MMP-2 and -9 were independent predictors of late recurrent carotid artery stenosis (p<0.03 and p<0.001, respectively). CONCLUSIONS: Higher plasma concentrations of active MMP-2 and -9 were associated with an increased risk of carotid restenosis with plaque recurrence. PMID- 19297220 TI - The long (and winding) road to gene discovery for canine hip dysplasia. AB - Hip dysplasia is a common inherited trait of dogs that results in secondary osteoarthritis. In this article the methods used to uncover the mutations contributing to this condition are reviewed, beginning with hip phenotyping. Coarse, genome-wide, microsatellite-based screens of pedigrees of greyhounds and dysplastic Labrador retrievers were used to identify linked quantitative trait loci (QTL). Fine-mapping across two chromosomes (CFA11 and 29) was employed using single nucleotide polymorphism (SNP) genotyping. Power analyses and preferential selection of dogs for ongoing SNP-based genotyping is described with the aim of refining the QTL intervals to 1-2 megabases on these and several additional chromosomes prior to candidate gene screening. The review considers how a mutation or a genetic marker such as a SNP or haplotype of SNPs might be combined with pedigree and phenotype information to create a 'breeding value' that could improve the accuracy of predicting a dog's hip conformation. PMID- 19297219 TI - Influence of CYP2C9 and VKORC1 on warfarin response during initiation of therapy. AB - BACKGROUND: Although multiple reports have documented the influence of CYP2C9 and VKORC1 variants on warfarin dose, risk of over-anticoagulation and hemorrhage, their influence on anticoagulation maintenance and individual proportion of time spent in target INR range (PPTR) is limited. Moreover the potential benefit of genotype-guided dosing implemented after initiation of therapy in a racially diverse population has not been explored. Herein we present the influence of CYP2C9 and VKORC1 C1173T on warfarin response during the first 30 days of therapy. METHODS: Warfarin dose was empirically determined in 250 African Americans 271 European Americans. The influence of CYP2C9 and VKORC1 on rate of INR increase, anticoagulation maintenance, risk of over-anticoagulation, and change in dose over 30 days was evaluated after adjustment for socio-demographic, lifestyle and clinical factors. Possession of variant VKORC1 (+/- variant CYP2C9) genotype was associated with a more rapid attainment of target INR and higher frequency of dose adjustments. Patients possessing variant genotypes spent less time in target range. However adjustment for rate of INR increase rendered the association non-significant. European Americans (but not African Americans) possessing variant VKORC1 (+/- variant CYP2C9) genotype had a higher risk of over anticoagulation. Neither CYP2C9 nor VKORC1 influenced the risk of minor hemorrhage. CYP2C9 and VKORC1 explained 6.3% of the variance in dose change over the first 30 days of therapy demonstrating that the usefulness of genotype-guided dosing may extend beyond first day of therapy. CONCLUSION: The benefit of genotype-based dose prediction may extend beyond first few days of therapy. Whether genotype-guided dosing will decrease the risk of over-anticoagulation, improve anticoagulation control and most importantly improve outcomes for chronic warfarin users remains to be proven. PMID- 19297221 TI - Lorazepam versus diazepam-phenytoin combination in the treatment of convulsive status epilepticus in children: a randomized controlled trial. AB - BACKGROUND: Convulsive status epilepticus demands urgent and appropriate management with anticonvulsants. Intravenous diazepam is an established drug in the management of convulsive status epilepticus in adults as well as in children. The efficacy of intravenous lorazepam has not been well established in children. OBJECTIVE: To determine whether intravenous lorazepam is as efficacious as diazepam-phenytoin combination in the treatment of convulsive status epilepticus in children. STUDY DESIGN: Randomized controlled trial. METHODS: A total of 178 children were enrolled in the study; 90 in the lorazepam group and 88 in the diazepam-phenytoin combination group. Enrolled subjects were between 1 and 12 years with a clinical diagnosis of convulsive status epilepticus, presenting in pediatric emergency of a tertiary care hospital. They were randomized to receive either intravenous lorazepam (0.1 mg/kg) or intravenous diazepam (0.2 mg/kg) phenytoin (18 mg/kg) combination at admission and were followed up for subsequent 18 h. RESULTS: The overall success rate of therapy was 100% in both the groups. There was no statistically significant difference in the two groups (lorazepam versus diazepam-phenytoin combination) in the median time taken to stop the seizure [20s in both groups], the number of subjects requiring more than one dose of the study drug to stop the presenting seizure [lorazepam 6(6.7%) versus diazepam-phenytoin combination: 14 (15.9%); adjusted RR (95% CI)=0.377 (0.377, 1.046); P=0.061] and the number (%) of patients having respiratory depression [lorazepam 4(4.4%) versus diazepam-phenytoin combination 5 (5.6%)]. None of the patients in the two groups required additional anticonvulsant drug to stop the presenting seizure. No patient required mechanical ventilation and none of the patients in the two groups required cross-over to the alternative regimen. CONCLUSION: Lorazepam is as efficacious and safe as diazepam-phenytoin combination. We recommend use of lorazepam as a single drug to replace the two drug combination of diazepam-phenytoin combination to control the initial seizure in pediatric convulsive status epilepticus. PMID- 19297222 TI - [Guide for the organisation of ceremonies for delivering pins and certificates of recognition to volunteer blood donors. Etablissement francais du sang Auvergne Loire. Associations pour le don du sang benevole]. AB - Award ceremonies constitute an important means of promotion among the blood donor population, and also the general population. They contribute to the development of blood donation loyalty. Even so, their organization must be rigorously and perfectly codified. With this aim, the Etablissement francais du sang Auvergne Loire, in partnership with the departmental representation of the blood donors national federation, worked out a guide for their associations. PMID- 19297223 TI - Prevalence and impact on quality of life of peripheral neuropathy with or without neuropathic pain in type 1 and type 2 diabetic patients attending hospital outpatients clinics. AB - AIMS: Diabetic polyneuropathy (DPN) without or with neuropathic pain (DPN-P) is one of the most frequent complications of diabetes. To better delineate their respective prevalences, we conducted a cross-sectional study that included 1111 patients (767 type 2 and 344 type 1 diabetic patients) followed up in diabetic outpatients clinics. The association of DPN and DPN-P with other diabetic complications, the impact on quality of life (QoL) and pain management were also investigated. METHODS: Two validated tools (Neuropen) and the DN4 questionnaire) were used to diagnose the two conditions. Pain intensity was measured using a visual analogue scale, and participants completed the 12-item Short-Form Health Survey to evaluate the physical and mental components of QoL. Univariate and multivariate models were used for the statistical analyses. RESULTS: The prevalence of DPN was 43% (95% CI 40.1-45.9), and was higher in type 2 (50.8%) than in type 1 (25.6%) diabetic patients. The prevalence of DPN-P was 14% (95% CI 12.1-16.2) which, again, was higher in type 2 (17.9%) than in type 1 (5.8%) patients. These prevalences both increased with age and diabetes duration. Nephropathy, obesity, low HDL cholesterol and high triglyceride levels were independently associated with DPN and/or DPN-P. Physical and mental components of QoL were significantly altered by DPN-P, but not DPN. Only half of the DPN-P patients were using analgesic treatment, while 28% were using anticonvulsants or antidepressants. CONCLUSION: DPN and DPN-P are frequent complications of diabetes, especially in type 2, and can be identified with inexpensive and easy to-use screening tools. Despite its profound impact on QoL, DPN-P remains undertreated. PMID- 19297224 TI - Low levels of insulin-like growth-factor-binding protein-1 (IGFBP-1) are prospectively associated with the incidence of type 2 diabetes and impaired glucose tolerance (IGT): the Soderakra Cardiovascular Risk Factor Study. AB - AIM: To explore the association between baseline levels of insulin-like growth factor-binding protein-1 (IGFBP-1), a marker of insulin sensitivity, and the development of type 2 diabetes or impaired glucose tolerance (IGT) in a specifically defined middle-aged population. METHODS: This cross-sectional population-based screening study was conducted in 1989-1990 and included baseline data for 664 non-diabetic subjects aged 40-59 years. Clinical data were collected and blood samples analyzed for blood glucose, serum lipids and insulin. Blood specimens were frozen at baseline and later analyzed for IGF-I, IGFBP-1 and C reactive protein (CRP). At the follow-up in 2006, the incidence of type 2 diabetes and IGT was reported based on primary-care medical records. RESULTS: During the 17-year observation period, 42 subjects (6.3%) developed type 2 diabetes/IGT. Those in the lowest quintile of IGFBP-1 (< or =24 microg/L) at baseline had a diabetes incidence of 12.6% while, in the highest quintile of IGFBP-1 (> or =59 microg/L), the incidence was 1.5%. Cox's proportional-hazards model regression analyses were used to determine the incidence of type 2 diabetes/IGT, corrected for age and gender, in relation to IGFBP-1, CRP and waist circumference. Subjects in the lowest IGFBP-1 quintile showed an independently increased risk of type 2 diabetes/IGT [hazards ratio (HR): 3.54; 95% CI 1.18 10.6; P=0.024]. For CRP and waist circumference, the corresponding figures were HR: 6.81; 95% CI 2.50-18.6; P<0.001 and HR: 3.33; 95% CI 1.47-7.6; P=0.004, respectively. CONCLUSION: Low levels of IGFBP-1 predicted the long-term development of type 2 diabetes or IGT in a middle-aged population. The association was independent of CRP and abdominal obesity. PMID- 19297225 TI - [Minutes of the 27th meeting of the European Society for Therapeutic Radiology and Oncology (ESTRO), Goteborg (Sweden), 14-18 September 2008, and of the 50th meeting of the American Society for Therapeutic Radiology and Oncology (ASTRO), Boston (USA), September 21-25 2008]. PMID- 19297226 TI - [Report of the mission commissioned by the Ministry of Health to the French Society of Radiation Oncology]. AB - Further to the Epinal events, Health Ministers P. Bas then R. Bachelot-Narquin have launched a plan of work devoted to radiotherapy; they have also committed the SFRO President, within the framework of a mission, to make proposals taking into account the demography of professionals and their level of competence, valorization of careers, cooperation with medical oncologists, delegations of authorities, mutualisation of human and material resources. Due to the numerous actions of the roadmap managed by the tutelages, the aim of the mission was focused on the modalities of work of the professionals linked to radiotherapy: radiation oncologists, radiographers and physicists. PMID- 19297227 TI - [An original contention system for hadron therapy]. AB - In hadron therapy centers that have only fixed horizontal beams (i.e. most carbon ions centers and protons centers of first generation), the angulations of the beam remain technically limited, especially for the treatment of children under general anaesthesia with posterior-oblique (40 degrees or so) beams in supine position. We have been developing recently an original positioning system allowing for treatment with posterior-oblique beams, either from right or left directions, by keeping the child in the adequate position. PMID- 19297228 TI - Fixed digital contractures revealing light-chain amyloidosis. AB - Among the many clinical manifestations of light-chain (AL) amyloidosis, musculoskeletal involvement is rarely reported. We describe the case of a 72-year old woman who was referred to our rheumatology department for fixed flexion contractures of the fingers that developed concomitantly with a decline in general health. Macroglossia and recent-onset dyspnea were noted. Investigations, which included a tongue biopsy, established the diagnosis of kappa light-chain amyloidosis with soft tissue, bone, and cardiac deposits. Melphalan and dexamethasone therapy was successful in stabilizing the clinical and laboratory abnormalities within 6 months. This case is remarkable in that the musculoskeletal manifestations were at the forefront of the clinical picture and led to the diagnosis. PMID- 19297229 TI - Prevalence and features of osteoporosis in the French general population: the Instant study. AB - OBJECTIVES: To determine the prevalence of diagnosed osteoporosis, the extent of treatment use and the incidence of fracture in a representative sample of the French general population. METHODS: A cross-sectional epidemiological survey of osteoporosis in 2613 women over 45 years in the general population was conducted using a stratified random sampling method and face-to-face interviews. Information was collected on the diagnosis of osteoporosis, fracture history, treatments, clinical and sociodemographic variables. Variables potentially associated with fracture were evaluated using stepwise multivariate logistic regression analysis. RESULTS: The overall prevalence of diagnosed osteoporosis was 9.7% [8.6%; 10.9%] and prevalence increased linearly with age. Overall, 155 women (61.0%) received osteoporosis treatment and treatment rates also increased with age. The most frequently prescribed treatments were bisphosphonates, in 50.3% of treated women. The treatment duration was over 2 years for 72.9% of treated women. Overall, 115 (45.3%) reported at least one previous fracture. Vertebral fractures were reported by 101 women (39.8%) and limb fractures by 41 women (16.1%). Multivariate logistic regression analysis identified fracture before the age of 40, menopause before the age of 40, use of sleeping pills, consultation with an eye specialist and history of cardiovascular disease as variables independently associated with fracture. CONCLUSIONS: Osteoporosis in France appears to be under-diagnosed and under-treated. Awareness and management of risk factors for osteoporosis and fracture could thus be improved. PMID- 19297230 TI - Validation of the KOFUS (Knee Osteoarthritis Flare-Ups Score). AB - OBJECTIVE: To develop a diagnostic score for knee osteoarthritis flare-ups and to evaluate its sensitivity and specificity. METHODS: We used two epidemiological databases built using the same methodology. One database was from a general practice study and served to develop the score, whereas the other was from a rheumatology study and served to validate the score. Physicians determined the flare-up status of each patient. The rheumatologist diagnosis was the reference standard. Logistic regression was performed to identify factors significantly associated with having a flare-up. RESULTS: Of the 6085 patients in the general practice database, 52.3% had a knee osteoarthritis flare-up. The score was built by assigning points to features that were present, with a weighting system based on the odds ratio of each feature for having a flare-up (0, feature absent; 1, morning stiffness for longer than 20 min; 2, pain causing nocturnal awakenings and knee effusion; 3, limping, joint swelling, and increased warmth over the knee). The score could range from 0 to 14. The receiver-operating characteristic curve showed that 7 was the best cutoff for diagnosing a flare-up. In the rheumatologist database, the numbers of patients having a flare-up were 274 (46.4%) based on the score and 270 (45.7%) based on the rheumatologist diagnosis. Sensitivity of the score was 87.0%, specificity 87.9%, positive predictive value 85.8%, and negative predictive value 89.0%. The Youden index was 0.75. CONCLUSION: A score equal to or greater than 7 points correlated well with a rheumatologist diagnosis of flare-up. Our score may constitute a valid objective criterion for standardizing the diagnosis of knee osteoarthritis flare-up, most notably when screening patients for inclusion in therapeutic trials. PMID- 19297231 TI - [Composite allotransplantation in the upper extremity: from research to clinical reality]. AB - Recent advancements in immunosuppression have made composite allotransplantation possible. To date, 38 hand allotransplantations have been performed in the world. Apart from the first case - amputated after voluntarily stopping the treatment- survival at 2-year follow-up has been 100%. After 2 years, 12 of the Chinese cases had to stop their treatment which led to complete loss of the transplanted parts. In 25 cases, results are known with a follow-up greater than 2 years. The transplanted patients all experienced one or more acute rejection episodes during the first postoperative year contrasting with only a 10% acute rejection rate following kidney transplantation. All these rejections were reversible with appropriate treatment. Extrinsic motor power was always restored, but intrinsic musculature only recovered in certain cases. Some sort of sensory discrimination was restored in 72% of the hands. All the patients had side effects related to immunosuppressive treatments, but none of these effects were life-threatening or impaired their quality of life. We think that composite allotransplantations should no longer be considered as clinical research and should be developed in accredited centres. PMID- 19297232 TI - Influence of the orientation of the Osstell transducer during measurement of dental implant stability using resonance frequency analysis: a numerical approach. AB - Stability of dental implants is measured by means of the Osstell device using a method of resonance frequency analysis. The aim of the study was to evaluate the role of direction-dependence of the Osstell transducer. For this purpose, a set of parametrical finite element analyses has been used. When rotating the transducer around the vertical axis the crucial phenomenon is the change in behavior of the system, which occurs in positions between 30 degrees and 80 degrees to the long axis of the alveolar crest. It seems from the presented results that, when measuring approximately perpendicularly to this long axis, the deviation from the ideal perpendicular position must not exceed 30 degrees. In this case, the first resonance frequency is recorded. When measuring in the approximately parallel position to the long axis of the alveolar crest, the second resonance frequency is recorded. The deviation from the ideal parallel position must not exceed 10 degrees. These values are not comparable with the previous ones. PMID- 19297233 TI - Image analysis of soft-tissue in-growth and attachment into highly porous alumina ceramic foam metals. AB - The detailed quantitative characterization of soft-tissue in-growth into highly porous artificial implants is critical to understanding the biophysical processes that will lead to the best structural scaffolding construct. Previous studies have performed mechanical peel tests and mostly qualitative histological analyses of soft-tissue. The goal of this paper is to report the results obtained from applying two image analysis algorithms to quantify the morphological structure found in histological images of stained soft-tissue in-growth into alumina ceramic foam metal implants using a canine model. Three different pore sizes were used and three different post-operative time points were considered. Using the 2D Wavelet Transform Modulus Maxima method and 2D Fourier Transform analysis, a strong anisotropic signature (directional preference) is detected in early (4 week) histological samples. The direction of preference is towards the center of the implants. The strength of the anisotropy at later time points (8 and 16 weeks) becomes gradually weaker. Our interpretation is that after a short period of time, the main tissue growth activity has been concentrated on filling the artificial implant by growing towards its center. The weaker anisotropic signature found at later time points is interpreted as the tissue growth activity strengthening its structure by growing in more random directions. PMID- 19297234 TI - Examining the interaction between food outlets and outdoor food advertisements with primary school food environments. AB - Schools are commonly seen as a site of intervention to improve children's nutrition, and prevent excess weight gain. Schools may have limited influence over children's diets; however, with home and community environments also exerting an influence within schools. This study considered the environment of food outlets and outdoor food advertisements surrounding four case study primary schools in New Zealand, and the impact of that external environment on within school food environments. The shortest travel route between school and home addresses, and the number of food outlets and advertisements passed on that route, was calculated for each student. Interviews with school management were conducted. The schools with a higher percentage of students passing food outlets and advertisements considered that their presence impacted on efforts within schools to improve the food environment. Limiting students' exposure to food outlets and outdoor food adverts through travel route planning, reducing advertising, or limiting the location of food outlets surrounding schools could be explored as intervention options to support schools in promoting nutrition. PMID- 19297235 TI - Ghrelin: a metabolic signal affecting the reproductive system. AB - Ghrelin, an acylated 28 amino acid gastric peptide, was isolated from the stomach as an endogenous ligand for growth hormone (GH) secretagogue receptor in 1999. Circulating ghrelin is mainly produced by specific cells in the stomach's oxyntic glands. Ghrelin potently stimulates GH release and food intake and exhibits diverse effects, including ones on glucose metabolism and on secretion and motility of the gastrointestinal tract. Besides these effects on food intake and energy homeostasis, ghrelin is also involved in controlling reproductive functions, and a role for it as a novel regulator of the hypothalamic-pituitary gonadal axis is clearly emerging. We review recent ghrelin research with emphasis on its roles in the reproductive axis. PMID- 19297236 TI - Enhancing the approach to selecting eyewear (EASE): a multi-centre, practice based study into the effect of applying contact lenses prior to spectacle dispensing. AB - PURPOSE: To examine the impact of contact lens application in non-contact lens wearers prior to spectacle dispensing. METHODS: Ninety-one eligible subjects with no previous contact lens experience were randomly assigned into either a test group (52 subjects) or control group (39 subjects). The former group was offered contact lenses before spectacle dispensing. The latter group proceeded to spectacle dispensing in the normal way without being offered contact lenses. The response to the offer of contact lenses, fit success, the impact on the spectacle dispensing process and the longer term purchase of contact lenses were measured and compared for the test and control groups. RESULTS: Of the test group, 46 (88%) agreed to try contact lenses. All subjects were suitable for contact lenses after a slit lamp examination and lenses were successfully fitted to 45 of the 46 subjects (98%). The test group reported a superior subjective spectacle dispensing experience and spent 267+/-143 pounds (mean+/-standard deviation) on their spectacle frames and lenses, 32% more than the control group (202+/-128 pounds). At 3 months post dispensing, 17 test patients (33% of original cohort) had purchased contact lenses compared with five control subjects (13%) (p=0.001). CONCLUSIONS: The opportunity to try contact lenses prior to spectacle dispensing was well received by subjects who generally reported a very positive experience. In particular, contact lenses assisted in seeing frame detail and suitability for wear. One third of subjects proceeded to purchase contact lenses. We recommend that practitioners consider offering contact lenses to all suitable patients who are proceeding to spectacle dispensing to optimise the dispensing process and to provide an opportunity to try contact lenses. PMID- 19297237 TI - Computational tools for designing and engineering biocatalysts. AB - Current computational tools to assist experimentalists for the design and engineering of proteins with desired catalytic properties are reviewed. The applications of these tools for de novo design of protein active sites, optimization of substrate access and product exit pathways, redesign of protein protein interfaces, identification of neutral/advantageous/deleterious mutations in the libraries from directed evolution and stabilization of protein structures are described. Remarkable progress is seen in de novo design of enzymes catalyzing a chemical reaction for which a natural biocatalyst does not exist. Yet, constructed biocatalysts do not match natural enzymes in their efficiency, suggesting that more research is needed to capture all the important features of natural biocatalysts in theoretical designs. PMID- 19297238 TI - Lessons from nature: unraveling biological CH bond activation. AB - The cleavage of unactivated CH bonds is one of the most challenging reactions in chemical biology. Metalloenzymes have evolved that efficiently perform these transformations with exquisite control of selectivity; however, a proposed requirement is the generation of highly reactive intermediates that could be lethal. A thermodynamic argument involving the putative reactive species is outlined, whereby the interplay between two tunable parameters, redox potential and pK(a), may be the key to sustainable function. In addition, factors that control these parameters are also described, including hydrogen-bonding networks found within protein active sites. Synthetic examples are used to corroborate these ideas. PMID- 19297239 TI - Anaerobic functionalization of unactivated C-H bonds. AB - The functionalization of alkanes was once thought to lie strictly within the domain of enzymes that activate dioxygen in order to generate an oxidant with suitable potency to cleave inert C-H bonds. The emergence of the radical SAM superfamily of enzymes-those which use S-adenosyl-l-methionine as a precursor to a 5'-deoxyadenosyl 5'-radical-has kindled a renaissance in the study of radical dependent enzymatic reactions, and is ushering in a wealth of new and intriguing chemistry that remains to be elucidated. This review will focus on a special subclass of radical SAM enzymes that functionalize inert C-H bonds, highlighting the functional groups and the chemistry that leads to their insertion. Within this class are first, enzymes that catalyze sulfur insertion, the prototype of which is biotin synthase; second, enzymes that catalyze P-methylation or C methylation, such as P-methylase or Fom3; third, enzymes that catalyze oxygen insertion, such as the anaerobic magnesium protoporphyrin-IX oxidative cyclase (BchE); and fourth, enzymes that functionalize n-hexane or other alkanes as the first step in the metabolism of these inert compounds by certain bacteria. In addition to surveying reactions that have been studied at various levels of detail, this review will speculate on the mechanisms of other types of reactions that this chemistry lends itself to. PMID- 19297240 TI - Synthesized some 4-(2-thiazolylazo)resorcinol complexes: characterization, thermal and optical properties. AB - Some transition metal complexes of the type [ML2].nH2O (n=0 or 2) of the title ligand, 4-(2-thiazolylazo)resorcinol, HL (TAR) have been synthesized and characterized by various analytical and physicochemical (elemental, thermal analyses, AAS, electrolytic conductance and magnetic susceptibility measurements) and spectral (UV-vis and IR) techniques for structure determination and optical properties. The complexes have the formulae [ML2] for M=Fe(II), Cu(II) and Zn(II); [CdL2].2H2O. An octahedral structure is proposed for all complexes. IR spectra show that the ligand is coordinated to the metal ions in a tridentate manner with ONN donor sites of the resorcinol OH, azo N and thiazole N. The effect of varying pH and solvent on the absorption behavior of the ligand and complexes has been investigated. The optical constants such as, refractive index, extinction coefficient, dielectric constant were determined for the ligand and its complexes. These parameters changed with different metal complexes. The optical absorption data revealed that the band gap of the films was direct transitions. The optical band gap and Urbach energy of the films were determined using the known theory. The optical dispersion parameters were determined according to Wemple and DiDomenico method. PMID- 19297241 TI - Theoretical study on local defect structure of (FeO4)(5-) clusters in YGG and LGG crystals. AB - The optical spectrum and the local defect structure of tetrahedral (FeO4)(5-) clusters are investigated in yttrium gallium garnet (YGG) and lutetium gallium garnet (LGG) crystals by means of the 252x252 complete energy matrices for d(5) configuration ions in tetragonal ligand field. The results show that the local defect structures around tetrahedral Fe3+ centers display an expansion effect. Simultaneously, the (FeO4(5-) clusters in the two different crystals have very similar local structures, which are close to those in YIG garnet crystal. Finally the relative curves of the zero-field splitting energies DeltaE1 and DeltaE2 in the ground state (6)A(1) varying with the structure parameter theta are plotted. PMID- 19297242 TI - Interaction of organotin with piperazine derived self-assembled cylindrical bisdithiocarbamates: spectral and thermal investigations. AB - Few organotin complexes of the type R4Sn2L2 where R=CH3 (2), C4H9 (3), C6H5 (4), and Sn2L2Cl4 (5) (L=bis(2,2'-dithiopiperazinato-2,2'-diaminodiethylamine)) have been synthesized and suitably characterized by FT-IR, UV-vis, 1H NMR, 119Sn NMR, ESI-MS, TGA/DSC, microanalysis and room temperature molar conductivity data. On the basis of FT-IR spectral studies, a symmetrical bidentate coordination has been proposed for all the complexes while the absence of any higher peak in the ESI-MS may be corroborated with the formation of binuclear complexes. On the basis of 119Sn NMR spectroscopy, six-coordinate geometry has been observed for the Sn-center in all the complexes. The TGA/DSC profile of the complexes implies their higher stability than its precursor. However, on the basis of IDT values the stability order of the organotin complexes was found to be 4>3>2. The room temperature conductivity values of the 1 mM solution of the ligand and its complexes are found to be comparable with that of non-ionic complexes. PMID- 19297243 TI - The profession of public health informatics: still emerging? AB - PURPOSE: Although public health informatics (PHI) was defined in 1995, both then and still now it is an "emerging" profession. An emergent profession lacks a base of "technical specialized knowledge." Therefore, we analyzed MEDLINE bibliographic citation records of the PHI literature to determine if a base of technical, specialized PHI literature exists, which could lead to the conclusion that PHI has emerged from its embryonic state. METHOD: A MEDLINE search for PHI literature published from 1980-2006 returned 16,942 records. Record screening by two subject matter experts netted 2493 PHI records that were analyzed by the intervals of previous PHI CBMs 96-4 and 2001-2 for 1980-1995 (I(1980)) and 1996 2000 (I(1996)), respectively, and a new, third interval of 2001-2006 (I(2001)). RESULTS: The distribution of records was 676 (I(1980)), 839 (I(1996)) and 978 (I(2001)). Annual publication rates were 42 (I(1980)), 168 (I(1996)), and 163 (I(2001)). Cumulative publications were accelerating. A subset of 19 (2.5%) journals accounted for 730 (29.3%) of the records. The journal subset average (+/ SD) annual publication rates of 0.7+/-0.6 (I(1980)), 2.9+/-1.9 (I(1996)), and 3.1+/-2.7 (I(2001)) were different, F(3, 64)=7.12, p<.05. Only I(1980) was different (p<.05) from I(1996) or I(2001). Average (+/-SE) annual rate of increase for all journals (8.4+/-0.8 publications per year) was different from the subset of 19 (2.7+/-0.3), t(36)=5.74, p<.05. MeSH first time-to-indexing narrowed from 7.3 (+/-4.3) years to the year (0.5+/-0.8) the term was introduced, t(30)=7.03, p<.05. CONCLUSION: A core set of journals, the proliferation of PHI articles in varied and numerous journals, and rapid uptake of MeSH suggest PHI is acquiring professional authority and now should not be tagged as an "emerging" profession. PMID- 19297244 TI - Epilepsy monitoring for therapy: challenges and perspectives. AB - The aim of the treatment of epilepsies is to control seizures without causing adverse effects in order to reach the best possible quality of life. Towards this aim diagnostic monitoring has become an increasingly important clinical tool, because seizures are transient events difficult to observe. Three major topics are discussed in this paper: (1) DIFFERENTIAL DIAGNOSIS: patients with difficult to treat epilepsies or unexplained paroxysmal events are referred to sophisticated monitoring units because they may have non-epileptic seizures or inaccurate classification of their epileptic seizures. (2) Evaluation of antiepileptic drugs: a prerequisite for the development of antiepileptic drugs it is desirable to employ objective measures to quantify efficacy, which can be achieved with the aid of diagnostic and therapeutic intensive seizure analysis including neuropsychological testing. (3) Automated computer supported seizure detection in emergency cases: for the quantification of frequency and severity of epileptic seizures in difficult to treat epilepsies (e.g. non-convulsive status epilepticus), adequate algorithms can be applied to long-term EEG recordings. These methods can be of assistance, as essential and practical tools, in the decision making during diagnostic monitoring by providing more objective results. PMID- 19297245 TI - Exploring the other side of cancer care: the informal caregiver. AB - OBJECTIVES: To identify the caregiver outcomes among relatives caring for patients with cancer and to examine the patient and family caregiver variables that predicted for caregiver burden and depression. METHODS: One hundred and thirty caregivers completed the Greek versions of the Burden Interview (BI), the Center of Epidemiology--Depression Scale (CES-D) and the Ways of Coping Questionnaire. Principal component analysis was performed to examine the underlying dimensions of caregiver outcome measures. One-way ANOVA and independent sample t-tests were used to test for differences in burden and depression in relation to demographic variables of interest. One-way ANOVA was used for examining differences in coping strategies. FINDINGS: One-way ANOVA showed that there are significant differences among the various educational levels (p<0.001) and the income (p<0.005) of the caregiver in terms of overall burden. 66.4% of caregivers had a depression above the usual cutoff point for depression. An independent samples t-test for possible gender differences, showed that there is a significant difference between males and females (p=0.29). In regression analysis it was found that only caregiver's income and patient's age are statistically significant in predicting burden and depression. When considering high-burdened caregivers results showed that there are significant differences in the use of coping strategies (p<0.001). CONCLUSIONS: Caregivers reported high levels of burden and depression. These outcomes of caregiving are related to several variables, but the caregiver's income and patient's age are predictive. Intervention strategies are needed to the vulnerable caregivers to help reduce burden and depression associated with caregiving. PMID- 19297246 TI - Evaluating the effects of the essential oils Leptospermum scoparium (manuka) and Kunzea ericoides (kanuka) on radiotherapy induced mucositis: a randomized, placebo controlled feasibility study. AB - This study evaluated the effects of an essential oil mouthwash on radiation induced mucositis of the oropharyngeal area during treatment for head and neck cancers. Nineteen adult patients completed the randomized placebo controlled trial which involved the use of a gargle containing 2 drops of a 1:1 mix of the essential oils of manuka (Leptospermum scoparium) and kanuka (Kunzea ericoides) in water. Those in the essential oil gargle group were observed to have a delayed onset of mucositis and reduced pain and oral symptoms relative to placebo (gargling with water) and the control ('usual care') groups. In addition those in the essential oil group were seen to have less weight loss (1% loss) than the other two groups (control 2.5%, placebo 4.5%). However a significant limitation in this study was the small sample size. Although the results from this feasibility study support the hypothesis that very small volumes of manuka and kanuka used in a gargle can provide a positive effect on the development of radiation induced mucositis, further research is required to confirm this finding. Randomization was applied according to the timing of the patient's entering the trial as well as their physical ability to gargle. Confirmation of these findings would pave the way for introduction of a simple, yet effective treatment for a condition which causes considerable discomfort and for which there is currently no definitive treatment. PMID- 19297247 TI - Mechanisms of hypoxic pulmonary vasoconstriction and their roles in pulmonary hypertension: new findings for an old problem. AB - Hypoxic pulmonary vasoconstriction (HPV) normally optimises ventilation-perfusion matching in the lung, but leads to pulmonary hypertension (PH) under conditions of global hypoxia. The past few years have provided some major advances in our understanding of this complex phenomenon, but significant controversy remains concerning many of the key underlying mechanisms. On balance, recent evidence is most consistent with an elevation in mitochondria-derived reactive oxygen species as a key event for initiation of HPV, with consequent Ca2+ release from intracellular ryanodine-sensitive stores, although the activation pathways and molecular identity of the associated Ca2+ entry pathways remain unclear. Recent studies have also raised our perception of the critical role played by Rho kinase (ROCK) in both sustained HPV and the development of PH, further promoting ROCK and the pathways regulating its activity and expression as important therapeutic targets. PMID- 19297249 TI - Paracetamol for stroke: easy in practice, but not in trials. PMID- 19297248 TI - The Paracetamol (Acetaminophen) In Stroke (PAIS) trial: a multicentre, randomised, placebo-controlled, phase III trial. AB - BACKGROUND: High body temperature in the first 12-24 h after stroke onset is associated with poor functional outcome. The Paracetamol (Acetaminophen) In Stroke (PAIS) trial aimed to assess whether early treatment with paracetamol improves functional outcome in patients with acute stroke by reducing body temperature and preventing fever. METHODS: In a multicentre, randomised, double blind, placebo-controlled trial, patients with ischaemic stroke or intracerebral haemorrhage and body temperature between 36 degrees C and 39 degrees C were randomly assigned treatment with paracetamol (6 g daily) or placebo within 12 h from symptom onset. Treatment allocation was based on a computer-generated list of random numbers with varying block size. The primary outcome was improvement beyond expectation on the modified Rankin scale at 3 months, according to the sliding dichotomy approach. This trial is registered, number ISRCTN74418480. FINDINGS: Between March, 2003, and May, 2008, 1400 patients were randomly allocated treatment. 260 (37%) of 697 patients receiving paracetamol and 232 (33%) of 703 receiving placebo improved beyond expectation (adjusted odds ratio [OR] 1.20, 95% CI 0.96-1.50). In a post-hoc analysis of patients with baseline body temperature 37-39 degrees C, treatment with paracetamol was associated with improved outcome (1.43, 1.02-1.97). There were 55 serious adverse events in the paracetamol group (8%) and 70 in the placebo group (10%). INTERPRETATION: These results do not support routine use of high-dose paracetamol in patients with acute stroke. Paracetamol might have a beneficial effect on functional outcome in patients admitted with a body temperature 37-39 degrees C, but this post-hoc finding needs further study. FUNDING: Netherlands Heart Foundation. PMID- 19297250 TI - Living with an unfixable heart: a qualitative study exploring the experience of living with advanced heart failure. AB - BACKGROUND: Nurses working with patients with advanced heart failure need knowledge that will help us to help patients cope with their situations of chronic illness. However, our knowledge bank is deficient due to the scarcity of inquiry that takes the affected person's point of view as its central focus. AIM: The aim of this study was to describe patients' experiences of living with advanced heart failure. METHODS: The study sample (N=9) consisted of male (N=6) and female (N=3) patients with advanced (NYHA classes III-IV) heart failure. The design was qualitative and open unstructured interviews were audio-taped and transcribed verbatim during 2006. RESULTS: Four main themes emerged: Living in the Shadow of Fear; Running on Empty; Living a Restricted life; and Battling the System. The experience of living with advanced heart failure was described as a fearful and tired sort of living characterised by escalating impotence and dependence. CONCLUSIONS: The findings suggest that there may be an illogical but enduring ethos of 'cure' pervading health care worker's attitudes to advanced heart failure care. This mindset might be working to hinder the application of additional or alternative therapies, which might better palliate the physical and psychosocial distress of patients. PMID- 19297251 TI - "Open" minimally invasive surgery in pediatric urology. AB - Minimally invasive surgery (MIS) offers alternative operative approaches to standard open surgical techniques. However, MIS has been defined primarily as substituting laparoendoscopic alternatives for the traditional open surgical approach. The concept of MIS methodology may also be applied to open surgery in an effort to decrease incision size, potentially reduce morbidity and enhance convalescence, without compromising 'gold standard' outcomes. Pediatric urological applications of open MIS include pediatric renal surgery, ureteral reimplantation, ureteral surgery, inguinal-scrotal and genital surgery. A thorough review of the pediatric urology literature was performed and studies were identified describing open MIS, including outcomes and complications. PMID- 19297252 TI - Macrophage activation syndrome: serological markers and treatment with anti thymocyte globulin. AB - Two patients presented at the University of Rochester Medical Center with a febrile illness, cytopenias, organ failure (liver failure or respiratory failure), and markedly elevated serum ferritin and sIL-2R. A diagnosis of probable macrophage activation syndrome was made. Both patients failed initial therapy with steroids and cyclosporine, either due to toxicity or lack of efficacy. Both patients responded dramatically to rabbit anti-thymocyte globulin (ATG). PMID- 19297253 TI - Role of paraventricular nucleus in exercise training-induced autonomic modulation in conscious rats. AB - The paraventricular nucleus (PVN) of the hypothalamus is an important site for autonomic regulation, where gamma-aminobutyric acid (GABA) system plays an important role. The central mechanisms underlying modulatory effects of exercise training have yet to be characterized. Our objective was to analyze the effects on the autonomic modulation and hemodynamic parameters after bicuculline or muscimol injections into the PVN of sedentary (control, C) and previously submitted to swimming training (ST) rats. After ST protocol, adult male Wistar rats, instrumented with guide cannulas to PVN and femoral artery and vein catheters were submitted to mean arterial pressure (MAP) recording. The exercise training reduced the LF oscillations in normalized units and increased the HF oscillations in absolute and normalized units. Compared with the C group, muscimol microinjections in the ST group promoted a higher decrease in MAP (C= 14+/-1 vs. ST=-28+/-4 mm Hg). Spectral analysis of HR (pulse interval) showed that the muscimol microinjections also reduced LF and HF oscillations in absolute units in both groups. Bicuculline microinjections increased the systolic arterial pressure (C=155+/-5, ST=164+/-5 mm Hg) in ST compared with the C group. Bicuculline injections also increased the LF oscillations of HR in absolute units in C and ST groups. Meanwhile, in normalized units only the ST group showed an increase in the LF oscillations. Our data showed that PVN has an important role in autonomic modulation after exercise training. PMID- 19297254 TI - Isocratic rapid liquid chromatographic method for simultaneous determination of carotenoids, retinol, and tocopherols in human serum. AB - An improved isocratic and rapid HPLC method was developed for the measurement of carotenoids, retinol and tocopherols in human serum. Vitamins were extracted with hexane. Mobile phase consisted of a mixture acetonitrile:methylene chloride:methanol with 20 mM ammonium acetate. This method used a small bead size (3 microm) Spherisorb ODS2 column with titane frits. Diode array and fluorescence detectors were used respectively for the detection of carotenoids and retinol/tocopherols. Chromatographic separation was complete in 13 min for beta cryptoxanthin, cis-trans-lycopene, alpha-carotene, beta-carotene, cis-beta carotene, retinol, delta-tocopherol, gamma-tocopherol and alpha-tocopherol. Echinenone and tocol were employed as internal standards for diode array and fluorescence detection. Accuracy was validated using standard reference material (SRM) 968C. Intra-assay and inter-assay precision were respectively 0.2-7.3% and 3.6-12.6%. Sensitivity was verified using the ICH recommendations and the limit of detection (LOD) obtained was sufficient for routine clinical application. PMID- 19297255 TI - Highly efficient peptide separations in proteomics. Part 2: bi- and multidimensional liquid-based separation techniques. AB - Multidimensional liquid-based separation techniques are described for maximizing the resolution of the enormous number of peptides generated upon tryptic digestion of proteomes, and hence, reduce the spatial and temporal complexity of the sample to a level that allows successful mass spectrometric analysis. This review complements the previous contribution on unidimensional high performance liquid chromatography (HPLC). Both chromatography and electrophoresis will be discussed albeit with reversed-phase HPLC (RPLC) as the final separation dimension prior to MS analysis. PMID- 19297256 TI - Determination of botulinum toxins after peptic sample pre-treatment by multidimensional nanoscale liquid chromatography and nano-electrospray ion-trap mass spectrometry. AB - Botulinum neurotoxin types A to G are produced from different strains of Clostridium botulinum. The complex neurotoxins belong to the most toxic substances known and cause botulism both in humans and animals. Botulinum toxin complexes are produced with molecular weights of 300, 500 and 900 kDa. These large protein complexes contain beside the toxic zinc protease of 150 kDa, additional neurotoxin associated proteins, which are responsible for the extreme pH and protease stability. In this study we present for the first time a rugged detection method of botulinum toxins at femtomole levels in complex culture media after peptic sample pre-treatment and 2D-nano-LC-ESI-MS-MS-technique. In contrast to other studies, we used progenitor toxins directly from culture supernatant of C. botulinum strains A, B, E and F without further purification, to simulate complex, protein-containing sample conditions. We were able to demonstrate, that peptic pre-treatment is a great challenge in reducing ubiquitous proteins as well as proteins from suspicious samples. The study also found that multidimensional chromatography leads to significant better peptide differentiation and identification in protein loaded matrices than one dimensional nano-LC-ESI-MS. PMID- 19297257 TI - Lactose malabsorption: Genetic testing versus lactose hydrogen breath test. PMID- 19297258 TI - Is CA 19-9 a screening marker? PMID- 19297259 TI - Reconstruction of goat femur segmental defects using triphasic ceramic-coated hydroxyapatite in combination with autologous cells and platelet-rich plasma. AB - Segmental bone defects resulting from trauma or pathology represent a common and significant clinical problem. In this study, a triphasic ceramic (calcium silicate, hydroxyapatite and tricalcium phosphate)-coated hydroxyapatite (HASi) having the benefits of both HA (osteointegration, osteoconduction) and silica (degradation) was used as a bone substitute for the repair of segmental defect (2 cm) created in a goat femur model. Three experimental goat femur implant groups- (a) bare HASi, (b) osteogenic-induced goat bone marrow-derived mesenchymal stem cells cultured HASi (HASi+C) and (c) osteogenic-induced goat bone marrow-derived mesenchymal stem cells cultured HASi+platelet-rich plasma (HASi+CP)--were designed and efficacy performance in the healing of the defect was evaluated. In all the groups, the material united with host bone without any inflammation and an osseous callus formed around the implant. This reflects the osteoconductivity of HASi where the cells have migrated from the cut ends of host bone. The most observable difference between the groups appeared in the mid region of the defect. In bare HASi groups, numerous osteoblast-like cells could be seen together with a portion of material. However, in HASi+C and HASi+CP, about 60-70% of that area was occupied by woven bone, in line with material degradation. The interconnected porous nature (50-500 microm), together with the chemical composition of the HASi, facilitated the degradation of HASi, thereby opening up void spaces for cellular ingrowth and bone regeneration. The combination of HASi with cells and PRP was an added advantage that could promote the expression of many osteoinductive proteins, leading to faster bone regeneration and material degradation. Based on these results, we conclude that bare HASi can aid in bone regeneration but, with the combination of cells and PRP, the sequence of healing events are much faster in large segmental bone defects in weight-bearing areas in goats. PMID- 19297260 TI - The ideal bone marking instrument in craniofacial cases. PMID- 19297261 TI - Thoraco-acromial artery perforator 'propeller' flap. PMID- 19297263 TI - Commentary on: Complications of otoplasty: a literature review. JPRAS 2009;62(1):19-27. PMID- 19297264 TI - Femoral deficiency reconstruction using a hydroxyapatite-coated locked modular stem. A series of 43 total hip revisions. AB - INTRODUCTION: We report a continuous prospective series of patients operated on for total hip prosthesis femoral component loosening involving a bone defect. Reconstruction was performed using a hydroxyapatite-coated locked modular stem. The study's objective was to assess medium term clinical and X-ray results obtained with this original concept. MATERIALS AND METHODS: The patients included received a REEF (DePuy) femoral implant for aseptic loosening or loosening associated with a periprosthetic fracture. Implantation was systematically accompanied by an extended trochanteric osteotomy (ETO). Patients were followed up prospectively by clinical and X-ray examination. Their loosening was graded at inclusion according to Vives' classification as revised by SOFCOT in 1999. Analysis focused on actuarial implant survivorship, dislocation and the bone/implant interface. RESULTS: Forty-three hips were included: mean follow-up was 58.2 months (12-92) and mean age at surgery was 72.4 years (37-94). The main indications were severe bone loss rated grade III (n=15) or IV (n=16) according to the SOFCOT classification. There was one long-term failure, involving implant fracture secondary to nonunion of the femoral shaft. Mean Postel and Merle d'Aubigne (PMA) clinical assessment score increased from six preoperatively to 14.5 at end of follow-up. X-ray analysis found no stem migration by end of follow up. There was consistent consolidation of the ETO around the stem, except in one case of stem fracture which evolved into tight nonunion. In terms of metaphyseal integration, five patients showed radiolucency without evolution over follow-up, and eight had severe calcar cortical atrophy at end of follow-up. Mean 5-year actuarial survivorship was 97.7+/-2.3%, with a 2% incidence of dislocation. DISCUSSION: The complications rate was low, and results were comparable with those reported in the literature. The study confirmed the interest of the extended trochanteric osteotomy exposure and the effectiveness of the hydroxyapatite-coated interlocked modular stem concept in the treatment of hip prosthesis loosening with femoral bone loss (involving or not the cortex). On analysis, the one case of failure does not prescribe the surgical technique employed. Implant osseointegration was difficult to analyze. The extent of the surgical approach and of its corresponding sequels, combined with the complexity of adjusting the implant, however, restrict this surgical option indications to level III and IV cases of femoral loosening. PMID- 19297265 TI - Frohse's arcade is not the exclusive compression site of the radial nerve in its tunnel. AB - INTRODUCTION: The radial tunnel is a musculo-aponeurotic furrow which extends from the lateral epicondyle of humerus to the distal edge of the supinator muscle. The superficial head of the supinator muscle forms a fibrous arch, the arcade of Frohse (AF), which is the most common site of compression of the radial nerve motor branch. The latter is less commonly compressed by the adjacent muscular structures. This tunnel syndrome might be worsened with repeated pronation and supination of the forearm. The double object of that work was: (1) to define the radial nerve anatomic landmarks, (2) to determine the anatomical relationship of the radial nerve main trunk and branches to the peripheral osseous and muscular structures in the anterior aspect of the elbow joint in order to identify which of these conflicting elements are likely to cause a compressive neuropathy. MATERIAL AND METHODS: The study design involved the dissection of 30 embalmed cadaveric upper limbs. Anatomic and morphometric investigations of the radial nerve, its terminal and motor branches were carried out. The presence of adhesions between radial nerve and joint capsule, tendons and aponeurotic expansions of epicondylar muscles and supinator arch was investigated. All measurements were taken in both pronation and supination of the forearm. RESULTS: Neither macroscopic radial compressive neuropathy at the level of the supinator arch nor adhesions between the radial nerve and the joint capsule were found. In four cases (13%), dense fibrous tissue surrounded the radial nerve supply to extensor carpi radialis brevis (ECRB). The fibrous arch of the supinator muscle arose in a semi-circular manner and was noted to be tendinous in 87% of the extremities and of membranous consistency in the remaining 13%. The length of the AF averaged 25.9 mm. The angle formed by the radial shaft and the supinator arch was 23 degrees. Neither fibrous structures nor adhesions of the deep branch of the radial nerve (DBRN) along its course through the supinator muscle were observed. DISCUSSION: Anatomic studies have revealed a variable rate of occurrence of a tendinous AF, which range from 30 to 80% (87% in our study) according to authors. It is reported to be a predisposing factor to the development of chronic entrapment neuropathy of the DBRN, especially if it is thick and provides a narrow opening for passage of the DBRN. The tendinous consistency of the supinator arch is believed to develop in adults, in response to repeated rotary movements of the forearm. Repetitive pronation and supination of the forearm induces compression of the radial nerve and its branches between two inextensible structures. The fibrous AF and the proximal end of the radius (radial head and radial tubercle). This condition is aggravated by the supinator muscle repeated activity. Repetitive compression might then promote histological changes in radial tunnel content and progressive development of a local fibrous zone. We also observed that the radial nerve supply to ECRB could be entrapped between the superolateral aspect of the ECRB and the superior edge of the supinator muscle. PMID- 19297266 TI - Bacterial biocide resistance. AB - The emergence of bacterial resistance to antimicrobial agents has caused increasing concern globally. the basis of bacterial resistance to antibiotics is well known, while the nonsusceptibility mechanisms of bacteria to biocides are less well understood. Recently, there is considerable interest in the problems associated with the development and spread of bacterial nonsusceptibility to biocides in order to understand the mechanisms of action and the bacterial nonsusceptibility mechanisms to biocides. Different groups of bacteria vary in their intrinsic nonsusceptibility to biocides, with bacterial spores being the most resistant, followed by mycobacteria, then Gram-negative organisms, with Gram positive bacteria generally being the most susceptible. this intrinsic nonsusceptibility in some instances might be associated with constitutive degradative enzymes or due to active efflux pumps, but in reality cellular impermeability is considered as a major factor that plays an important role in the emergence of bacterial nonsusceptibility to biocides. Nonsusceptibility associated with biofilmforming bacterial cells can be considered an intrinsic nonsusceptibility mechanism resulting from physiological (phenotypic) adaptation cells. PMID- 19297267 TI - Antimicrobial susceptibility among bacterial isolates from ICU and non-ICU setting and different age groups: results from the tigecycline evaluation and Surveillance trial in North America. AB - As part of the tigecycline evaluation and Surveillance trial (t.e.S.t.), isolates were collected from centers in North America. In vitro activity was assessed for isolates collected in 2006 using ClSi guidelines and ClSi or FDA interpretive criteria. Data were analyzed according to intensive care unit (iCU) or non-iCU location and age group (18-64 years and > 65 years).Rates of mRSA in 2006 were higher among isolates from patients aged >65 years (52.0%) than younger patients (48.4%) and from non-iCU settings (48.0%) than from the iCU (45.3%). Rates of vancomycin-resistant E. faecium, multidrug-resistant Acinetobacter spp. (resistant to levofloxacin and amikacin as well as the carbapenems and/or third generation cephalosporins), and eSbl-producing K. pneumoniae among isolates collected from the iCU were 70.7%, 5.8%, and 16.6%, respectively.Tigecycline, linezolid and vancomycin were active against all isolates of S. aureus, including mRSA, from both settings and the two age groups. Among E. faecium, >95% of isolates were susceptible to linezolid, and tigecycline maintained MIC(90)s of 0.06-0.12 mg/l. Against Acinetobacter spp., the most active antimicrobials were tigecycline and minocycline irrespective of age group or patient setting (MIC(90)s 2 and 8 mg/l, respectively). Percentage susceptibility off K. pneumoniae was >90% against tigecycline, imipenem, meropenem, cefepime and amikacin for isolates from both age groups and settings. Against eSbl-producing K. pneumoniae, imipenem (88.9%-96.4%) and tigecycline (85.1%-100%) demonstrated the highest rates of activity. tigecycline demonstrated excellent activity against clinically relevant resistant organisms. PMID- 19297268 TI - Determining in vitro synergistic activities of tigecycline with several other antibiotics against Brucella melitensis using checkerboard and time-kill assays. AB - Antimicrobial therapy of Brucella spp. infection is difficult because there are relatively few effective treatment regimens, and single-agent therapy has generally been considered inadequate due to unacceptably high relapse rates. tigecycline, the first in a new class of antimicrobials, the glycylcyclines, is a 9-t-butylglycylamido derivate of minocycline. in this study, the in vitro activity of tigecycline in combination with gentamicin, streptomycin, rifampin, co-trimoxazole, levofloxacin, and minocycline was investigated using the checkerboard method to evaluate 16 Brucella melitensis isolates. The time-kill method was used to determine the bactericidal activities of combinations of tigecycline with rifampin, gentamicin, and levofloxacin, which were found (via the checkerboard method) to have a synergistic effect in combinations with tigecycline. Using the checkerboard method, combinations of rifampin, gentamicin, and levofloxacin with tigecycline showed synergistic effects against 5 (31.2%), 3 (18.9%), and 8 (50%) of the isolates. No synergy was observed with tigecycline in combination with minocycline, streptomycin, or co-trimoxazole. tigecycline with gentamicin achieved the earliest complete killing at 4x miC (in 6 h), while complete killing with the other combinations was delayed up to 24 h. the time kill method showed that the combination of tigecycline and levofloxacin had an antagonistic effect, while the checkerboard method detected synergy and no interaction effects. these data suggest that a combination regimen of tigecycline with gentamicin and rifampin may be a good choice for treating brucellosis. PMID- 19297270 TI - Difference in killing activity of caspofungin and paradoxical growth between Candida albicans and C. krusei clinical isolates in different media. AB - Minimum fungicidal concentration (mFC) of caspofungin was determined against 16 Candida albicans and 16 C. krusei in Rpmi-1640 and antibiotic medium 3 (Am3). time-kill tests were performed on six C. albicans and four C. krusei strains at 0.06-16 mg/l caspofungin. mFC ranges after 48 h were 0.5-1 and 1-2 mg/l for C. albicans and C. krusei, respectively; one C. albicans and the C. krusei reference strain showed paradoxical growth (pG) in Rpmi-1640, respectively. in killing experiments, after 48 h caspofugin was fungicidal against two and four C. albicans in Rpmi-1640 (at 16 mg/l) and in Am3 (at >0.5 mg/l), respectively; pG was noted in three and two cases, respectively. Caspofungin at >2 and 0.5 mg/l was fungicidal against all tested C. krusei strains even after 24 h in Rpmi-1640 and Am3, respectively. Killing activity of caspofungin against C. albicans and C. krusei could be exactly measured only by killing curves. PMID- 19297269 TI - Antimicrobial susceptibility patterns and characterization of clinical isolates of Staphylococcus aureus in immunocompromised cancer patients undergoing chemotherapy in Pakistan. AB - Cancer patients undergoing chemotherapy are highly prone to infections because of suppression of their immune system. in the present study, the activity of fluoroquinolones, cephalosporins, glycopeptides and oxazolidinones was evaluated in Staphylococcus aureus strains by broth dilution method according to Clinical and laboratory Standards institute (ClSi), USA guidelines. the frequency of methicillin-resistant S. aureus (mRSA) and methicillin-sensitive S. aureus (mSSA) strains was 67% and 33% respectively. The MIC(50 )and MIC(90 )of isolates were ciprofloxacin 8 and 32 microg/ml for mRSA, 8 and 16 microg/ml for mSSA, levofloxacin 8 and 16 microg/ml for mRSA and mSSA, gatifloxacin 4 and 16 microg/ml for mRSA and 4 and 8 microg/ml for mSSA. MIC(50 )and MIC(90 )of vancomycin and linzolid were 1 and 2 microg/ml, 2 and 4 microg/ml for both mRSA and mSSA strains respectively. this study shows a high prevalence of and resistance in mRSA. However, vancomycin and linezolid were highly active and could be used for treating mRSA infections. PMID- 19297271 TI - Pharmacist-driven aminoglycoside quality improvement program. AB - Our objective was to determine the impact of a pharmacist-driven aminoglycoside quality improvement program on the dosing methods of aminoglycosides at our institution. We assessed our current quality through retrospective review of all patients receiving aminoglycosides during a 5-month period. we then developed and implemented a pharmacist-driven aminoglycoside dosing program and prospectively assessed patients during a matched 5-month period. two hundred and sixteen patients were evaluated, 87 pre-program and 129 post-program. Prior to standardized pharmacist intervention, 44% of patients achieved optimal therapy. Oost implementation, patients achieving optimal therapy increased to 80% (p < 0.001). Patients in the pre-program group had a higher rate of acute changes in renal function compared to the post-program group (14.9% versus 6.2% [p <0.05]). This pharmacist-driven aminoglycoside dosing quality improvement program increased the frequency of optimal dosing of aminoglycosides and reduced the incidence of nephrotoxicity at our institution. PMID- 19297272 TI - The impact of total control of antibiotic prescribing by infectious disease specialist on antibiotic consumption and cost. AB - We analyzed the impact on antimicrobial consumption of antimicrobial therapy totally guided by an infectious disease specialist (iD). in a teaching hospital antibiotic consumption was recorded yearly from 1998 to 2005 in three units. Antibiotic policy was introduced in 1999. in Unit A, restricted antimicrobials were prescribed after approval by the head of the unit. the iD was involved as a consultant upon request. in unit b, restricted antimicrobials were approved by an iD. All other antimicrobials were prescribed by the physicians on the ward. in unit C, all antimicrobials were prescribed by an iD. A significant decreasing trend in antibiotic consumption in defined daily doses (DDD) per patient day and per admission, and cost of antimicrobials per patient was observed in unit C, a decreasing trend in antibiotic consumption per patient was observed in unit b as well. totally iD guided antimicrobial therapy is an efficient method of antimicrobial policy. PMID- 19297273 TI - Clinical evaluation after a notification policy of linezolid use: a case series of 22 patients. AB - Linezolid exhibits a broad spectrum of activity against Gram-positive cocci, including Methicillin-resistant Staphylococcus aureus (mRSA) and vancomycin resistant enterococci (VRe). However, recent studies have already reported the emergence of linezolid-resistant mRSA or VRe. the purpose of this study is to evaluate not only the efficacy of linezolid for the treatment of nosocomial mRSA infections but also the effect of a notification policy of linezolid use. the charts of inpatients who had been treated with linezolid were reviewed for clinical outcome. After introduction of the notification policy of linezolid use, the clinical success rate was 73.3%, and the rate of appropriate linezolid use was 80%, whereas the success rate was 14.2% and the appropriate use rate was 14.3% before the policy. in conclusion, appropriate use controlled by a notification policy of antibiotics use is essential for prevention of the emergence and spread of linezolid-resistant bacteria, and for proper demonstration of its antibacterial ability. PMID- 19297274 TI - Late onset ventilator-associated pneumonia due to multidrug-resistant Acinetobacter spp.: experience with tigecycline. AB - The aim of the study was to evaluate the clinical success rate of 73 patients with ventilator-associated pneumonia (VAP) caused by multidrug-resistant (MDR) Acinetobacter spp. treated with tigecycline in seven intensive Care Units in Argentina and to determine which predictor variables were significant in this context. Clinical success in our patients was 69.86% (Ci= 58.65-81.07%) 51/73, without significant differences between patients with VAP due to MDR Acinetobacter spp. carbapenem-susceptible or carbapenem-resistant and only susceptible to colistin, minocyline and tigecycline (70% 44/73 vs. 69% 29/73 respectively, p=0.9006), and between patients who received <48h of prior antibiotics (including those who did not receive any) and those who received >48h of prior antibiotics (73.3% 22/30 vs 67.4% 29/43 respectively, p=0.7791). Age >67 and using other method than BAL for respiratory sampling were identified as predicting variables for negative clinical outcome. Our results suggest that tigecycline may be an acceptable alternative for therapy in patients with VAP caused by MDR-Acinetobacter spp. Nevertheless, only controlled clinical trials will provide the evidence to support approval for new indications. PMID- 19297275 TI - The significance of Chlamydia trachomatis in urethritis and prostatitis - differences in therapeutic approach - Croatian experience. AB - We examined a total of 1014 patients over 18 years of age; 252 with urethritis and 762 with chronic prostatitis syndrome. the mean age of patients with urethritis was 32.7 and with prostatitis syndrome 37.6 years. Clinical symptoms of urethritis were present from a few days to several months. in patients with chronic prostatitis syndrome, symptoms were present for at least 3 months. Chlamydia trachomatis alone was confirmed in 26 (10%) and in combination with Ureaplasma urealyticum in 6 (2%) patients with urethritis. in 171 (68%) patients with urethritis neither C. trachomatis nor U. urealyticum or Mycoplasma hominis were found. C. trachomatis alone was confirmed in 70 (9%), and in combination with other microorganisms in 7 (1%) patients with chronic prostatitis syndrome. in Croatia, the frequency of chronic chlamydial prostatitis has not significantly changed in the last 10 years, while the frequency of infections among adolescents decreased. the recommended regimen for acute chlamydial urethritis in Croatia is azithromycin 1.0 g as a single dose, and a total dose of 4-4.5 g azithromycin for chronic chlamydial prostatitis. PMID- 19297276 TI - Switch therapy in full-term neonates with presumed or proven bacterial infection. AB - This case-control study of full-term newborns with presumed or proven bacterial infection compared the efficacy, safety and tolerability of switch antibiotic therapy and traditional completely intravenous antibiotic administration. there were 36 newborns treated with switch therapy (i.v. ampicillin + sulbactam combined with i.v. amikacin for 3 days followed by oral cefpodoxime proxetil for 5 days); there were 72 full-term newborns with the same characteristics as controls who received i.v. ampicillin + sulbactam combined with i.v. amikacin for 3 days followed by i.v. ampicillin + sulbactam alone for a further 5 days. the results showed that full-term newborns with presumed or proven bacterial infection initially treated with intravenous antibiotics can be switched to oral antibiotics after 3 days' therapy if physical and laboratory data indicate the disappearance of infection, thus significantly reducing the length of stay in the neonatal intensive care unit and significantly increasing breastfeeding, without having any negative clinical impact. PMID- 19297277 TI - DNA repair proteins and telomerase reverse transcriptase in the cochlear lateral wall of cisplatin-treated rats. AB - Cochlear lateral wall damage is a side effect of cisplatin chemotherapy. Recent studies have shown that cisplatin treatment precipitates platinated DNA adducts in the cochlear lateral wall which suggest that DNA damage may contribute to ototoxicity. Platinated adducts are high-affinity substrates for the global genomic nucleotide excision repair (GG-NeR) pathway which is facilitated by xeroderma pigmentosum (Xp) complementing proteins, such as XpC, XpD and XpA. tumor biology has shown that in addition to stimulating GG-NeR, cisplatin may deplete telomerase reverse transcriptase (teRt). in the current study Fischer344 rats were treated with cisplatin (2 mg/kg/4 days, i.p.) and their cochleae harvested for immunohistochemistry. XpC, XpD and XpA expression increased while teRt expression decreased among cisplatin treated animals compared to vehicle control. these findings suggest that in addition to forming platinated adducts, cisplatin chemotherapy may up-regulate DNA repair proteins and modify teRt expression in the cochlear lateral wall. PMID- 19297278 TI - A phase I study of TS-1 plus carboplatin in patients with advanced non-small-cell lung cancer. AB - TS-1 is a novel oral anticancer agent comprised of tegafur, a prodrug of 5- flurouracil, and two modulators. A phase i study of tS-1 plus carboplatin combination therapy was conducted to determine the maximum tolerated dose (MTD), recommended dose (RD), and dose limiting toxicities (DLT) in advanced non-small cell lung cancer (NSClC). TS-1 was given orally at a dose of 80 mg/m(2)/day for 2 weeks, followed by a 2-week rest. Carboplatin was given intravenously on day 8 at a dose of 4.0, 5.0, 6.0 area under the curve (AUC) values. Fifteen patients with advanced NSClC were analyzed. the grade 3-4 toxicities observed during the first cycle were febrile neutropenia (6%), anemia (6%), anorexia (6%), and diarrhea (6%). these toxicities were reversible and manageable. The MTD for carboplatin was evaluated to be more than 6.0 AUC values, as one of six patients developed Dlt at this dose. the RD for carboplatin was estimated as 6.0 AUC values. Objective responses were seen in five patients (response rate 33%). PMID- 19297279 TI - Lipoplatin monotherapy: A phase II trial of second-line treatment of metastatic non-small-cell lung cancer. AB - Lipoplatin is a liposome encapsulated form of cisplatin. phase i studies on lipoplatin have demonstrated that the compound has an excellent toxicity profile. therefore we performed a phase ii trial in heavily pre-treated patients with advanced non-small-cell lung cancer (NSClC), performance status 0-2 in which the primary endpoint was response rate and secondary endpoints were safety and overall survival.Nineteen patients, average age 64 years old, with stage iV NSClC, were treated with lipoplatin 100 mg/m(2) every two weeks, as second line chemotherapy.We observed 1 partial remission (5.2%) and 3 stable diseases (15.9%). Time to progression (ttp) was 16 weeks and median overall survival (OS) was 31 weeks (7.2 months). We observed G1-G2 toxicity during chemotherapy, mainly gastrointestinal with nausea and vomiting (4 patients), asthenia (3 patients), mucositis (2 patients) and anemia (4 patients).Our phase ii study does not support a more extensive use of lipoplatin in phase III studies. An increase of dosage and a better selection of patients are mandatory to understand the real therapeutic activity of lipoplatin. PMID- 19297280 TI - Intra-arterial chemotherapy with gemcitabine and cisplatin for patients with recurrence of transitional cell carcinoma confined to the pelvis. AB - Post-cystectomy recurrence of transitional-cell carcinoma (TCC) that is confined to the pelvis is uncommon, and few data exist to guide its management. we used weekly intra-arterial chemotherapy with gemcitabine and cisplatin in a consecutive series of 11 patients with this diagnosis. After 2 cycles of intra arterial chemotherapy with gemcitabine (each cycle: 900 mg/m(2) on days 1, 8, and 15) plus cisplatin (each cycle: 30 mg/m(2) on days 1, 8, and 15), 3 patients achieved complete response (CR) and 8 patients partial response (PR). two pR patients achieved CR after surgery and radiotherapy for residual disease. Six pR patients died of disease progression 8 to 20 months after chemotherapy. Four CR patients have survived, free of disease, 17 to 43 months, and 1 CR patient died of a non-tumor-related cause 23 months after chemotherapy. Short course intra arterial chemotherapy with a GC regimen thus appears to be highly effective in the subset of patients with recurrent TCC confined to the pelvis. PMID- 19297281 TI - Emergence of a plasmid mediated bla (VIM-1) in Citrobacter koseri: report from the SENTRY Antimicrobial Surveillance Program (Italy). PMID- 19297282 TI - Parenteral administration of tobramycin for pulmonary exacerbations in cystic fibrosis patients: toxicity, serum levels and efficacy. PMID- 19297283 TI - Breakthrough Candida dubliniensis fungemia in an acute myeloid leukemia patient during voriconazole therapy successfully treated with caspofungin. PMID- 19297284 TI - Voriconazole for cryptococcal meningitis in children with leukemia or receiving allogeneic hemopoietic stem cell transplant. PMID- 19297285 TI - Combination treatment for chronic HCV hepatitis in lymphangioleiomyomatosis: first case report. PMID- 19297286 TI - [Toxoplasma gondii serology in pregnant woman: characteristics and pitfalls]. AB - Screening program of congenital toxoplasmosis depends principally on serological testing and follow-up of pregnant women with negative serology from the onset of pregnancy. This program is particularly efficient because the majority of cases of obstetrical seroconversion are diagnosed and followed up. Serological techniques have evolved with time and are many on the market. The "IgG" tests are more and more specific at the detriment of their sensitivity. The latter is variable from one technique to the other. In 2008, despite the presence of an international standard, serological titrations are particularly variable making interpretation of the kinetics impossible between two different series, two techniques or two laboratories. In cases of interpretation discordance or borderline values, it is useful to do a confirmatory technique. The dye test or the western blot are useful in these cases and prevent up to 2 to 8% of unnecessary obstetric follow up. IgM tests are becoming more and more sensitive and precocious. These tests frequently detect "non specific IgM" as well as "residual IgM" (associated with a stable positive IgG titre). In such cases and in the absence of a preceding positive result, a complementary test done by specialized laboratories (such as ISAgA IgM or IgA, IgG avidity test, differential agglutination, etc.) is indispensable in order to prevent any error of interpretation. These complementary tests as well as the serological evolution (control done in 15 days) permit, in a large number of cases, to reassure the patient with full confidence. PMID- 19297287 TI - [Primary distal renal tubular acidosis]. AB - Renal tubular acidosis are forms of metabolic acidosis characterized by an impairment of urinary acidification due to a lack of urine excretion of protons or loss of bicarbonates. Primary distal renal acidosis (dRTA) is characterized by hyperchloremic metabolic acidosis due to failure in proton excretion, variably severe nephrocalcinosis and/or nephrolithiasis associated with hypercalciuria and hypocitraturia. When the metabolic acidosis is compensated, dRTA can be diagnosed by the failure of urinary acidification after oral ammonium chloride or furosemide administration. dRTA is inherited as either an autosomal dominant or autosomal recessive trait. An autosomal dominant form results from a SLC4A1 gene mutation leading to dysfunction of the anionic exchanger type 1 (AE1). Otherwise, recessive forms are due to mutations of ATP6V1B1 gene encoding the B1-subunit of H+-ATPase expressed in the apical membrane of the alpha intercalated cells in collecting duct and in the cochlea. Those mutations lead to dRTA accompanied by sensorineural deafness. Also, mutations in ATP6V0A4 gene encode the accessory subunit a4 of the H+ATPase, leading to recessive forms of dRTA with preserved hearing or delayed signs of deafness. Molecular approach can identify mutations which are responsible for this pathology. The medical treatment is simple and involves an alkali load which allows curing the metabolic acidosis. Long-term outcome is usually good unless the patient's compliance is low or alkalizing treatment is insufficient. PMID- 19297288 TI - [The role of biology in the diagnosis of cerebrospinal fluid leaks]. AB - Cerebrospinal fluid leakage is a rare but critical condition with a substantial risk of intracranial infection, therefore its diagnosis and treatment is of major importance. CSF leakage diagnostic can be a challenging problem. Nephelometric measurement of beta-trace protein in the liquorrhoea is a non-invasive and fast method that can be used for CSF leakage diagnosis. It should kept in mind, however, that the cut-off of 1.1 mg/L is not suitable for patients with bacterial meningitis and those with a reduced glomerular filtration rate. Complementary use of beta-trace protein assay and beta2-transferrin detection is therefore recommended. PMID- 19297290 TI - [A critical study of the screening practices of gestational diabetes of a service of gynecology and obstetrics]. AB - Our study has for aim to describe the practices of screening of the gestational diabetes mellitus. Screening test was realised at 61% of the women included in the study. The HGPO in 50 g was prescribed in 47% of the cases, a single fasting blood glucose in 28% of the cases; 13,4% of the detected women had a glycaemia included between 1,30 and 2 g/L one hour after a load of 50 g of glucose. Among these patients, 52,63% did not realize test HGPO in 100 g. Gestational diabetes mellitus complicates about 7,7% of pregnancies in our study. In statistical analysis united varied, are considered as risk factors: the age, the family history of diabetes and the histories of macrocosmic during previous pregnancy. PMID- 19297289 TI - [Carbonyl stress and oxidatively modified proteins in chronic renal failure]. AB - Oxidative stress is commonly observed in chronic renal failure patients resulting from an unbalance between overproduction of reactive oxygen species and impairement of defense mechanisms. Proteins appear as potential targets of uremia induced oxidative stress and may undergo qualitative modifications. Proteins could be directly modified by reactive oxygen species which leads to amino acid oxydation and cross-linking. Proteins could be indirectly modified by reactive carbonyl compounds produced by glycoxidation and lipo-peroxidation. The resulting post-traductional modifications are known as carbonyl stress. In addition, thiols could be oxidized or could react with homocystein leading to homocysteinylation. Finally, tyrosin could be oxidized by myeloperoxidase leading to advanced oxidative protein products (AOPP). Oxidatively modified proteins are increased in chronic renal failure patients and may contribute to exacerbate the oxidative stress/inflammation syndrome. They have been involved in long term complications of uremia such as amyloidosis and accelerated atherosclerosis. PMID- 19297291 TI - Determination of urinary trans, trans-muconic acid reference values in general Tunisian population. AB - We wish to determine the urinary trans,trans-muconic acid reference values in the Tunisian general population, and evaluate the impact of several factors (age, gender, tobacco...) on these reference values. Urine samples were collected from 182 healthy Tunisian subjects who had not been occupationally exposed to benzene. This determination was performed by solid phase extraction sampling technique together with high performance liquid chromatography-photodiode array detector. Trans,trans-muconic values ranged from 0.003 to 0.618 mg/g creatinine, the 95% reference interval was: 0.004-0.36, and the 90% confidence interval of the upper reference limit was: 0.24-0.62 mg/g creatinine. Urinary trans,trans-muconic levels were significantly higher among smokers. Significant differences were also observed for the < 20 and the > or = 40 age groups. As a result urinary trans,trans-muconic background levels allow the biomonitoring of workers occupationally exposed to benzene at levels as low as 0.5 ppm. Age and tobacco, but not gender may affect the trans,trans-muconic reference values. PMID- 19297292 TI - [Study of association of the SNP19 polymorphism of calpain 10 gene with type 2 diabetes in ethnic sub-groups of the Tunisian population: gene-environment interaction]. AB - Calpain 10 (CAPN10) is the first diabetes gene to be identified through a genome scan followed by positional cloning, encoding the cysteine protease, the calpain 10 encodes for a ubiquitously expressed protease implicated in the two fundamental pathophysiological aspects of T2DM insulinoresistance and insulinosecretion. Many investigators, but not all, have subsequently found association between calpain 10 polymorphism and type 2 diabetes (T2DM) as well as insulin action and insulin secretion. The aim of this study was to determine whether there is an association between specific polymorphism SNP19 in CAPN10 gene and T2DM in two ethnic groups from Djerba Island. Overall, 162 patients with type 2 of diabetes and 110 healthy volunteers who served as controls for genetic characterization with no family history of diabetes were included in the present study. They consisted of 159 women and 113 men. Their mean +/- SD age was 56,47 +/- 11,86 years. All subjects were genotyped according to SNP 19 polymorphism in CAPN10 gene with PCR method to perform case-control study. After adjusting for gender and age, we found an association with a high risk of T2DM in Djerba Island only in Arab sub-group. PMID- 19297293 TI - [Patient identification vigilance in public healthcare organizations: inventory in French hospitals and proposals]. AB - Utilization of patients databases by the different healthcare departments, technical teams and registration clerks in care centers is becoming more and more important. Patients data exchanges within and between centers are growing, including public and private structures networks and require a complete match of the patient identification data. Because clinical chemists and pathologists are deeply involved in these exchanges, the National Clinical Chemistry College designed a survey about french hospitals practices (2007 july-2008 august) in order to secure patients identification at any stage of their stay. "Identity vigilance" term tends to emerge for designating this new risk management field. The responses from 94 hospitals are analyzed, which demonstrate an heterogeneous implementation and a very unequal concern of this issue in the different sites; 18% have not yet initiated any action. A strong involvement of the pathologists in the applying of identification safety procedures is expressed by their presence in the three executive, operative and underling levels in more than half of sites (in 84% of the sites for the operative level). Only 17% of the pathologists consider the patients database of their hospital software to be of good quality. This survey leads to become aware of local and national tasks that have to be implemented or carried on, both for identity registration at admission and identity checking before any health care. This effort has to be supported by an essential institutionalization of identity vigilance. PMID- 19297294 TI - [Epidemiological retrospective survey intestinal parasitism in the Provincial Hospital Center (Kenitra, Morocco): review of 10 years (1996-2005)]. AB - The survey drew up the epidemiological situation of intestinal parasitism in the center of health El Idrissi (Kenitra, Morocco). The number of reviews has decreased between 1996 and 2005. A correlation between the number of examinations and years of the study period was observed (p <0.001). 4285 stool specimens collected in 1996-2005 were tested by parasitologic examination. Among the persons examined, 606 of them were parasited by one or several species, say an infestation index of 14.15%. Amoeba were frequently observed (47.04%) with prevalence of Entamoeba histolytica (23.74%), followed by Flagella (28.79%) represented by: Giardia intestinalis (22.71%), Trichomonas intestinalis (5.49%) and Chilomastix mesnilii (0.60%). Helminthes were less found. Ascaris lumbricoides was frequent among helminthes (11.87%), followed by Trichuris trichiura (5.64%), Hymenolepis nana (2.68%), Enterobius vermicularis (2.08%), Taenia saginata (0.75%) and Stronyloides stercoralis (0.45%). The clinical symptoms were observed in 110 subjects with parasites (110/606 or 18.15%) characterized by abdominal pain (75 cases) and association diarrhea more abdominal pain (35 cases). The relationship between the infestation index calculed, sex, age, the annual and seasonal changes, polyparasitism and intestinal parasitic infection is discussed. PMID- 19297295 TI - [Urine Dipstick Screening tests: can they allow to conclude the absence of a urinary tract infection in diabetic hospitalised patients]. AB - Asymptomatic urinary tract infections are common in diabetic patients. The aim of this 10 months prospective study is to evaluate urinary dipstick tests versus cytobacteriological examination to conclude the absence of urinary tract infection in diabetic subjects. Each diabetic patient hospitalised for less than 8 hours and for whom it was decided a cytobacteriological examination was included in the study (141 samples). At the same time (and at patient's bedside) a dipstick urinalisys (glucose, leucocytes, nitrite, blood, protein, and ketone) was carried out. Sensitivity, specificity, negative predictive value, post test probability and negative likehood ratio were calculated. RESULTS: the combination of leucocyte zone with nitrite zone (both negative) has a 85,2% sensitivity, avoids 65% of cytobacteriology, but has an odd ratio at 0,20. Those results are improved when the glucose zone (negative test or less than 4 crosses) is taken into account, with a 96,3% sensitivity, 63,4% cytobacteriology avoided and a negative likehood ratio at 0,06. CONCLUSION: The addition of the glucose test to the usual leucocytes and nitrite tests seems to allow one to conclude the absence of urinary tract infection in diabetic patients; this is worth studying with a more extensive sample. PMID- 19297296 TI - [Difficulties of interpretation in toxoplasma serology in a pregnant woman in the context of a maternofetal alloimmunization against HPA-1a platelet antigen]. AB - Serological status in case of Toxoplasma gondii infection needs to be established either before or at the beginning of a pregnancy. However, clinical biologists are often facing conflicting serological results that are difficult to interpret: we report here the case of a woman in her 30th week of pregnancy. Both her IgM and IgG were negative at the 14th week of pregnancy; but suddenly, starting from the 20th week, her IgG became positive while her IgM remained negative. We remind here of the most frequent hypothesis that can explain a sudden and isolated increase of anti-T. gondii IgG: Is it a technical problem (specificity)? Is it a drug interference? Eventually, we found that the patient was receiving, since the 16th week of pregnancy, every week an intravenous perfusion of polyvalent immunoglobulins. Since we didn't know if the IgG present in this perfusion can protect the patient against toxoplasmosis, we decided to consider this women as non immune. PMID- 19297297 TI - [Multidrug intoxication]. AB - The authors report an suicide attempt by a 43 years old woman treated by Previscan, Effexor, Rivotril, and Stilnox for phlebitis anteriority and anxious depressive disorders. On arrival to the emergency unit, the medical status and the patient history required a cardiovascular reanimation and gastric washing. The haemostasis analysis demonstrates an anticoagulant overdose and the prescription of vitamine K1 and PPSB was necessary. Later, the multidrug poisoning was demonstrated thanks to blood, urinary and gastric fluid high performance liquid chromatography toxicological analyses. This case report points out that biological and toxicological analyses are complementary. The first ones are useful to determine the emergency symptomatic cares, and to check the drugs' efficiency. The second ones, although longer, are necessary to identify with certainty incriminated drugs. PMID- 19297298 TI - [Glossary of the acronyms for the institutions concerned by metrology]. AB - Metrology and one of its contributions, metrological traceability represent two fundamental developments for the clinical laboratories. Several international and national institutions take part in these developments. They elaborate recommendations going from concepts to implementation in the clinical laboratories. These activities are helpful for accrediting clinical laboratories. PMID- 19297299 TI - Unmet needs of patients with systemic lupus erythematosus. AB - The authors' goal was to assess unmet needs of patients with systemic lupus erythematosus (SLE). Participants (N = 112), who were recruited through the mailing list and support group meetings of a Lupus Alliance of America Affiliate, completed a survey based on prior research. All participants perceived at least 1 unmet need. The most frequently reported unmet needs were in the physical symptoms domain. Older patients were more likely than younger patients to have higher levels of unmet needs related to physical and psychological functioning. African American patients were more likely than white patients to have higher levels of unmet needs related to health services and information. Our findings document the high prevalence and variety of unmet needs among these patients, as well as variations among demographic groups. To address unmet needs of SLE patients, targeted referrals to patient educators, mental health professionals, and support organizations are important adjuncts to medical treatment. PMID- 19297300 TI - A comparison of smokers' and nonsmokers' fruit and vegetable intake and relevant psychosocial factors. AB - The authors examined the relation between smoking status and fruit and vegetable (FV) consumption among a population-based sample and examined differences in psychosocial factors that may influence diet and inform intervention efforts. The authors recruited adults (N = 2,540) from 5 US health plans to participate in a Web-based dietary intervention trial. At baseline, smokers ate fewer FV servings per day (p < .001) and were less likely to meet the 5 A Day goal (p < .001). Smokers reported lower self-efficacy, overall motivation, and intrinsic motivation for meeting daily FV recommendations. Fewer smokers expected that eating 5 FV servings a day would reduce their risk for diabetes (p = .02) or obesity (p = .008). Smokers are an important target group for dietary intervention. Intervention efforts should attempt to increase smokers' motivation and confidence in their abilities to change their eating patterns and educate them about the health benefits of eating FV. PMID- 19297301 TI - Understanding nonsmoking in African American and Caucasian college students: an application of the theory of planned behavior. AB - Few studies have considered whether psychological determinants of nonsmoking among college students vary by ethnicity. The authors tested the theory of planned behavior (TPB) to explain differences in nonsmoking intentions of 238 African American and 197 Caucasian college students who completed an in-class TPB questionnaire and a smoking assessment 1 week later. After removing 35 students who reported smoking at the baseline assessment, regressions were used to examine ethnic effects on TPB constructs when predicting nonsmoking intentions. Caucasians had statistically significant higher nonsmoking intentions than African Americans. Further, subjective norms for Caucasians and attitudes for African Americans had small but significant relations to intention, but perceived behavioral control (PBC) was a strongly significant predictor for both ethnic groups. However, the prediction of nonsmoking intentions was not statistically moderated by ethnicity for any of the TPB constructs. This study suggests that the TPB may aid in understanding collegiate nonsmoking intentions and help begin to explain differences in smoking on the basis of ethnicity. Last, because of strong associations shown in this study, PBC should be considered when developing ethnic-specific smoking interventions in college students. PMID- 19297302 TI - Mediated effects of social support for healthy nutrition: fruit and vegetable intake across 8 months after myocardial infarction. AB - The authors investigated relationships among family support for healthy nutrition, fruit and vegetable intake, and social-cognitive mediators such as intention, self-efficacy, and planning. Participants were 130 myocardial infarction survivors (64% men) treated in 4 clinics in Poland. The authors collected self-reported data across 8 months during and after cardiac rehabilitation (at the beginning of rehabilitation, 2 weeks after rehabilitation, and 6 months later). Results indicate that directly after rehabilitation, less than 20% of patients met recommended guidelines for fruit and vegetable intake, with a further decrease in fruit and vegetable consumption over time (14% and 12% at follow-ups). Family support predicted patients' behavior at follow-up, with self-efficacy playing a mediating role. Although regular formation of action plans predicted fruit and vegetable intake, it did not mediate social support behavior relationships. PMID- 19297303 TI - Uncompensated care cost: a pilot study using hospitals in a Texas county. AB - The financial ramifications of uncompensated care cost (UCC) on the healthcare industry have been difficult to quantify. With the lack of a standardized definition of uncompensated care and the need to account for the uninsured, indigent, and immigrant populations, the authors identified $190 million of UCC from Southwestern border hospitals for emergency room treatment of undocumented immigrants and $934 million of uncompensated care charges for 23 hospitals in a Texas county, which translated to $353 million of UCC. Although lawmakers passed the Medicare Prescription Drug Improvement and Modernization Act (2003) to address the growing imbalance, the shortfall of funds highlights the growing crisis and need for policy intervention. PMID- 19297304 TI - Geographic variation in minority participation in hospital management in the United States. AB - Minority participation in hospital management is, by all estimates, lower than almost anyone is prepared to accept. However, to date, there has been no published study of hospital management diversity using comprehensive national data. In the present study, the authors used data from the U.S. Equal Employment Opportunity Commission to estimate more directly and comprehensively how management diversity in private U.S. hospitals varies by geographic area. Although minorities constitute over 33% of the U.S. population, they constitute only 14% of all top-level managers and officials in private U.S. hospitals. There is enormous geographic variation in this average across cities and regions. The authors found that minority management participation is systematically related to market factors, including demographic and education variables. These systematic patterns explain about 90% of the Metropolitan Statistical Area differences in minority manager percentage. However, even after adjusting observed variations for these differences, the authors found important unexplained geographic variations in minority managerial employment in hospitals across U.S. cities. PMID- 19297305 TI - Healthcare consumers' beliefs about medical malpractice lawsuits. AB - In the United States, malpractice tort reform has come to be regarded by many as a principal means of reducing the high costs of medical care. Despite growing popular sentiment for tort reform evidenced in public opinion polls and in earlier research, the authors wondered about the experiences and feelings that enhance this pro-malpractice-tort-reform position. The authors report the results of their comparative inquiry into consumers' beliefs that there are too many medical malpractice lawsuits. The authors analyze data derived from a nonprobability sample of consumer residents of West Virginia and of international residents of Houston, Texas, completed in 2007-2008. Only a small percentage (about 5%) of the consumer respondents reported having filed a medical malpractice claim. Of those who did file, about 70% agreed with the position that there are too many medical malpractice lawsuits filed each year. Consumers who reported that they did not feel that a lawsuit had much of an impact on a physician's practice or lifestyle were less inclined to also feel that too many medical malpractice lawsuits are filed. The authors also found evidence that feeling that there are too many medical malpractice lawsuits bore a weak, inverse relation to the belief that medical errors have been on the rise over the last 10 years. PMID- 19297306 TI - Interview: healthcare industry players. Interview by Dennis S. Palkon and Oliver Baranczyk. PMID- 19297307 TI - Depression and C-reactive protein: population-based Health 2000 Study. AB - OBJECTIVE: To test whether depression is independently associated with elevated levels of C-reactive protein (CRP) in a large nationwide population-based sample. METHODS: Depression and CRP were assessed in 6005 Finns aged >30 years, as part of the ongoing population-based Health 2000 Study. Depression was determined by responses to the Beck's Depression Inventory (BDI-21) and by Composite International Diagnostic Interview (CIDI). RESULTS: Higher scores on BDI-21 were related to higher CRP levels (beta = 0.12, p < .001) in men and (beta = 0.08, p < .001) in women. In men, but not in women, this relationship persisted after adjustment for a number of other known risk factors. In men, the more recent dysthymic disorder or at least moderate depressive episode defined using CIDI was related to elevated CRP levels (beta = 0.06, p = .006). CONCLUSIONS: Depressive symptoms may be partly responsible for inflammatory processes, and inflammatory processes may induce depressive symptoms in men. PMID- 19297308 TI - Caregiving for children with developmental disabilities is associated with a poor antibody response to influenza vaccination. AB - OBJECTIVE: Older spousal caregivers of dementia patients have been found to show a relatively poor antibody response to medical vaccination. In the present case control study in a different caregiving environment, we sought to compare antibody responses to vaccination in parents of children with disabilities and parents of typically developing children. METHODS: At baseline assessment, 32 parents of children with developmental disabilities and 29 parents of typically developing children completed standard measures of perceived stress and child problem behaviors. They also provided a blood sample and were then vaccinated with the thymus-dependent trivalent influenza vaccine. Further blood samples were taken at 1- and 6-month follow-ups. RESULTS: Relative to parents of typically developing children (mean titer = 458, standard deviation (SD) = 155.7 at 1 month follow-up and mean titer = 265, SD = 483.0 at 6-month follow-up), caregivers (mean titer = 219, SD = 528.4 at 1-month follow-up and 86, SD = 55.0 at 6-month follow-up) mounted a poorer antibody response than controls to the B/Malaysia strain of the vaccine. CONCLUSION: The negative impact of caregiving on antibody response to vaccination would not seem to be restricted to older spousal caregivers, but is also evident in younger parents caring for children with developmental disabilities. The behavioral characteristics of the care recipients may be a determinant of whether or not antibody response to vaccination is compromised. PMID- 19297309 TI - Lower ambulatory blood pressure in chronic fatigue syndrome. AB - OBJECTIVE: To examine blood pressure circadian rhythm in subjects with chronic fatigue syndrome (CFS) and appropriate normal and fatigued controls to correlate parameters of blood pressure regulation with perception of fatigue in an observational cohort study. The cause of CFS remains unknown and there are no effective treatments. METHODS: To address whether inactivity was a confounder, we performed a 24-hour ambulatory blood pressure monitoring in the following three subject groups: 1) CFS patients (Fukuda Diagnostic criteria) (n = 38); 2) normal controls (n = 120); and 3) a fatigue comparison group (n = 47) with the autoimmune liver disease primary biliary cirrhosis (PBC). All patients completed a measure of fatigue severity (Fatigue Impact Scale). In view of the different demographics between the patient groups, patients were age- and sex-matched on a case-by-case basis to normal controls and blood pressure parameters were compared. RESULTS: Compared with the control population, the CFS group had significantly lower systolic blood pressure (p < .0001) and mean arterial blood pressure (p = .0002) and exaggerated diurnal variation (p = .009). There was a significant inverse relationship between increasing fatigue and diurnal variation of blood pressure in both the CFS and PBC groups (p < .05). CONCLUSION: Lower blood pressure and abnormal diurnal blood pressure regulation occur in patients with CFS. We would suggest the need for a randomized, placebo-controlled trial of agents to increase blood pressure such as midodrine in CFS patients with an autonomic phenotype. PMID- 19297310 TI - Diagnostic accuracy of predicting somatization from patients' ICD-9 diagnoses. AB - OBJECTIVES: To hypothesize in a new and different population that administrative database (ADB) screening would identify somatizing patients by increasing numbers of visits, female gender, and greater percent of International Classification of Diseases, 9th Edition (ICD-9) primary diagnosis codes in musculoskeletal, nervous, gastrointestinal (GI), and ill-defined body systems. We labeled these codes as having "somatization potential." Our earlier study demonstrated that ICD 9 codes and other data from the ADB effectively identified somatization. METHODS: Using a prospective observational design in a staff model health maintenance organization, we evaluated 1364 patients aged 18 to 65 years who had > or =8 visits yearly in the 2 years before study. Clinician raters applied a reliable method of medical chart review to identify patients meeting the criteria for somatization. We randomly selected 2/3 for the derivation set (n = 901) for logistic regression to evaluate the contribution of potential ADB correlates (age, gender, all encounters, primary diagnosis codes (ICD-9), revenue codes, and charges) of a diagnosis of somatization. This prediction rule was then applied to the remaining 1/3 of subjects, the validation set (n = 463). RESULTS: Patients averaged 47.1 years, 12.8 visits per year, and 71.6% were female; 319 had somatization. Age, visits, and somatization potential were associated with clinician-rated somatization, with a c-statistic 0.72 in the derivation set and 0.68 in the validation set. CONCLUSIONS: These data support our earlier findings that selected ICD-9 diagnoses in the ADB predict somatization, suggesting their potential in identifying a common, costly, and usually unrecognized problem. PMID- 19297311 TI - Depression and whole blood serotonin in patients with coronary heart disease from the Heart and Soul Study. AB - OBJECTIVE: To evaluate whether depression is associated with whole blood serotonin in outpatients with stable coronary heart disease (CHD). Depression is associated with incident CHD and with adverse cardiovascular outcomes. Dysregulation of peripheral serotonin, common to both depression and CHD, may contribute to this association. METHODS: We performed a cross-sectional study of 791 participants with stable CHD enrolled in the Heart and Soul Study and not taking antidepressant medication. We assessed major depression using the Computerized Diagnostic Interview Schedule (CDIS-IV) and measured whole blood serotonin (WBS) from fasting venous samples. RESULTS: Of the 791 participants, 114 (14%) had current (past month) major depression, 186 (24%) had past (but not current) major depression, and 491 (62%) had no history of depression. Age adjusted mean WBS was higher in participants with current major depression (139 +/- 6.5 ng/ml) than in those with past depression (120 +/- 5.0 ng/ml) or no history of depression (119 +/- 3.1 ng/ml) (p = .02). This association was unchanged after adjustment for demographic characteristics, medical comorbidities, medication use, and cardiac disease severity (p = .02). When serotonin was analyzed as a dichotomous variable, current depression was associated with a 70% greater odds of having WBS in the highest quartile (adjusted odds ratio = 1.71; 95% Confidence Interval = 1.03-2.83; p = .04). CONCLUSIONS: In this sample of patients with stable CHD, current major depression was independently associated with higher mean WBS levels. Future studies should examine whether elevated WBS may contribute to adverse outcomes in patients with depression and CHD. PMID- 19297312 TI - Predicting risk of type 2 diabetes in England and Wales: prospective derivation and validation of QDScore. AB - OBJECTIVE: To develop and validate a new diabetes risk algorithm (the QDScore) for estimating 10 year risk of acquiring diagnosed type 2 diabetes over a 10 year time period in an ethnically and socioeconomically diverse population. DESIGN: Prospective open cohort study using routinely collected data from 355 general practices in England and Wales to develop the score and from 176 separate practices to validate the score. PARTICIPANTS: 2 540 753 patients aged 25-79 in the derivation cohort, who contributed 16 436 135 person years of observation and of whom 78 081 had an incident diagnosis of type 2 diabetes; 1 232 832 patients (7 643 037 person years) in the validation cohort, with 37 535 incident cases of type 2 diabetes. OUTCOME MEASURES: A Cox proportional hazards model was used to estimate effects of risk factors in the derivation cohort and to derive a risk equation in men and women. The predictive variables examined and included in the final model were self assigned ethnicity, age, sex, body mass index, smoking status, family history of diabetes, Townsend deprivation score, treated hypertension, cardiovascular disease, and current use of corticosteroids; the outcome of interest was incident diabetes recorded in general practice records. Measures of calibration and discrimination were calculated in the validation cohort. RESULTS: A fourfold to fivefold variation in risk of type 2 diabetes existed between different ethnic groups. Compared with the white reference group, the adjusted hazard ratio was 4.07 (95% confidence interval 3.24 to 5.11) for Bangladeshi women, 4.53 (3.67 to 5.59) for Bangladeshi men, 2.15 (1.84 to 2.52) for Pakistani women, and 2.54 (2.20 to 2.93) for Pakistani men. Pakistani and Bangladeshi men had significantly higher hazard ratios than Indian men. Black African men and Chinese women had an increased risk compared with the corresponding white reference group. In the validation dataset, the model explained 51.53% (95% confidence interval 50.90 to 52.16) of the variation in women and 48.16% (47.52 to 48.80) of that in men. The risk score showed good discrimination, with a D statistic of 2.11 (95% confidence interval 2.08 to 2.14) in women and 1.97 (1.95 to 2.00) in men. The model was well calibrated. CONCLUSIONS: The QDScore is the first risk prediction algorithm to estimate the 10 year risk of diabetes on the basis of a prospective cohort study and including both social deprivation and ethnicity. The algorithm does not need laboratory tests and can be used in clinical settings and also by the public through a simple web calculator (www.qdscore.org). PMID- 19297313 TI - Screening for type 2 diabetes in primary care. PMID- 19297314 TI - Docetaxel plus oblimersen sodium (Bcl-2 antisense oligonucleotide): an EORTC multicenter, randomized phase II study in patients with castration-resistant prostate cancer. AB - BACKGROUND: This randomized, phase II study assessed the activity of oblimersen sodium, a Bcl-2 antisense oligonucleotide, administered before docetaxel (Taxotere) to patients with castration-resistant prostate cancer. PATIENTS AND METHODS: Chemotherapy-naive patients with prostate-specific antigen (PSA) progression and testosterone < or = 0.5 ng/ml received docetaxel 75 mg/m2 on day 1 or oblimersen 7 mg/kg/day continuous i.v. infusion on days 1-7 with docetaxel 75 mg/m2 on day 5 every 3 weeks for < or = 12 cycles. Primary end points were confirmed PSA response (Bubley criteria) and major toxic events. RESULTS: Confirmed PSA response was observed in 46% and 37% of 57 and 54 patients treated with docetaxel and docetaxel-oblimersen, respectively. Partial response (RECIST) was achieved in 18% and 24%, respectively. Oblimersen added to docetaxel was associated with an increase in the incidence of grade > or = 3 fatigue, mucositis, and thrombocytopenia. Major toxic events were reported in 22.8% and 40.7% of patients with docetaxel and docetaxel-oblimersen, respectively. CONCLUSIONS: The primary end points of the study were not met: a rate of confirmed PSA response >30% and a major toxic event rate <45% were not observed with docetaxel-oblimersen. PMID- 19297315 TI - The NER proteins are differentially expressed in ever smokers and in never smokers with lung adenocarcinoma. AB - BACKGROUND: The expression levels of excision repair cross-complementation group 1 (ERCC1), replication protein A (RPA) and xeroderma pigmentosum group F (XPF) nucleotide excision repair proteins may be important in the response to platin based therapy in lung cancer patients. It is not known whether ERCC1, RPA and XPF expression levels differ between ever smokers (ES) and never smokers (NS). PATIENTS AND METHODS: ERCC1, RPA and XPF expression levels were immunohistochemically evaluated in 125 patients with resected lung adenocarcinoma (AC) and carefully reviewed smoking status. RESULTS: ERCC1 was correlated with XPF (P = 0.001), but not with RPA (P = 0.11). In the univariate analysis, ERCC1 and XPF levels were higher in NS compared with ES (P = 0.004 and P = 0.003, respectively). In the multivariate analysis, the smoking status was predictive of the ERCC1 level [odds ratio (OR) 2.5, 95% confidence interval (CI) 1.03-6.2] after adjustment for variables linked to the smoking status, including age and the presence of bronchioloalveolar (BAC) features. The smoking status was also predictive of both RPA (OR 6.7, 95% CI 1.5-33.3) and XPF levels (OR 12.5, 95% CI 2.9-50) after adjusting for age, sex and BAC features. CONCLUSION: In patients with resected lung AC, ERCC1, RPA and XPF expression levels are higher in NS compared with ES. PMID- 19297316 TI - Early detection of second breast cancers improves prognosis in breast cancer survivors. AB - BACKGROUND: The impact of early detection of second breast cancers in women who have survived a primary breast cancer is unknown. We examined the prognostic effect of detection of ipsilateral breast relapse (IBR) or contralateral breast cancer (CBC) in the asymptomatic relative to symptomatic phase. PATIENTS AND METHODS: Subjects were women with histology-verified second (invasive or in situ) breast cancer (N = 1044) in a breast centre in Florence (1980-2005). Symptom status, test, tumour stage, and outcomes data were obtained from clinical records and linkage with mortality registry. Disease-specific survival was measured from first cancer diagnosis to avoid lead-time bias. Sensitivity analysis was used to allow for length-time bias. RESULTS: Second cancers (IBR = 455; CBC = 589; median age 60 years) were diagnosed in 699 asymptomatic and 345 symptomatic women (67% versus 3%, P < 0.0001). Mammography was more sensitive than clinical examination (86% versus 57%, P < 0.0001); however, 13.8% of cases were only identified clinically. Asymptomatic cancers were smaller than symptomatic for both IBR (P < 0.001) and CBC (P < 0.001). Early-stage tumours were more frequent in asymptomatic (58.1%) than symptomatic (22.6%) women (P < 0.0001). Fewer women with asymptomatic than symptomatic CBC had node metastases (P = 0.0001). Hazard ratio (HR) for asymptomatic (relative to symptomatic) detection was 0.51 (0.32 0.80) for IBR, 0.53 (0.36-0.78) for CBC, and 0.53 (0.40-0.72) in all subjects (P < 0.0001). Length bias-adjusted HRs ranged from 0.53 to 0.73. CONCLUSION: Detection of second breast cancers in the asymptomatic phase leads to detection of early-stage cancer and improves relative survival by between 27% and 47%. PMID- 19297317 TI - Modulation of the oligomerization state of p53 by differential binding of proteins of the S100 family to p53 monomers and tetramers. AB - We investigated the ways S100B, S100A1, S100A2, S100A4, and S100A6 bind to the different oligomeric forms of the tumor suppressor p53 in vitro, using analytical ultracentrifugation and multiangle light scattering. It is established that members of the S100 protein family bind to the tetramerization domain (residues 325-355) of p53 when it is uncovered in the monomer, and so binding can disrupt the tetramer. We found a stoichiometry of one dimer of S100 bound to a monomer of p53. We discovered that some S100 proteins could also bind to the tetramer. S100B bound the tetramer and also disrupted the dimer by binding monomeric p53. S100A2 bound monomeric p53 as well as tetrameric, whereas S100A1 only bound monomeric p53. S100A6 bound more tightly to tetrameric than to monomeric p53. We also identified an additional binding site for S100 proteins in the transactivation domain (1-57) of p53. Based on our results and published observations in vivo, we propose a model for the binding of S100 proteins to p53 that can explain both activation and inhibition of p53-mediated transcription. Depending on the concentration of p53 and the member of the S100 family, binding can alter the balance between monomer and tetramer in either direction. PMID- 19297318 TI - Inhibitory mechanism of Escherichia coli RelE-RelB toxin-antitoxin module involves a helix displacement near an mRNA interferase active site. AB - In Escherichia coli, RelE toxin participates in growth arrest and cell death by inducing mRNA degradation at the ribosomal A-site under stress conditions. The NMR structures of a mutant of E. coli RelE toxin, RelE(R81A/R83A), with reduced toxicity and its complex with an inhibitory peptide from RelB antitoxin, RelB(C) (Lys(47)-Leu(79)), have been determined. In the free RelE(R81A/R83A) structure, helix alpha4 at the C terminus adopts a closed conformation contacting with the beta-sheet core and adjacent loops. In the RelE(R81A/R83A)-RelB(C) complex, helix alpha3(*) of RelB(C) displaces alpha4 of RelE(R81A/R83A) from the binding site on the beta-sheet core. This helix replacement results in neutralization of a conserved positively charged cluster of RelE by acidic residues from alpha3(*) of RelB. The released helix alpha4 becomes unfolded, adopting an open conformation with increased mobility. The displacement of alpha4 disrupts the geometry of critical residues, including Arg(81) and Tyr(87), in a putative active site of RelE toxin. Our structures indicate that RelB counteracts the toxic activity of RelE by displacing alpha4 helix from the catalytically competent position found in the free RelE structure. PMID- 19297319 TI - Superoxide-mediated formation of tyrosine hydroperoxides and methionine sulfoxide in peptides through radical addition and intramolecular oxygen transfer. AB - The chemistry underlying superoxide toxicity is not fully understood. A potential mechanism for superoxide-mediated injury involves addition to tyrosyl radicals, to give peptide or protein hydroperoxides. The rate constant for the reaction of tyrosyl radicals with superoxide is higher than for dimerization, but the efficiency of superoxide addition to peptides depends on the position of the Tyr residue. We have examined the requirements for superoxide addition and structurally characterized the products for a range of tyrosyl peptides exposed to a peroxidase/O(2)(.) system. These included enkephalins as examples of the numerous proteins and physiological peptides with N-terminal tyrosines. The importance of amino groups in promoting hydroperoxide formation and effect of methionine residues on the reaction were investigated. When tyrosine was N terminal, the major products were hydroperoxides that had undergone cyclization through conjugate addition of the terminal amine. With non-N-terminal tyrosine, electron transfer from O(2)(.) to the peptide radical prevailed. Peptides containing methionine revealed a novel and efficient intramolecular oxygen transfer mechanism from an initial tyrosine hydroperoxide to give a dioxygenated derivative with one oxygen on the tyrosine and the other forming methionine sulfoxide. Exogenous amines promoted hydroperoxide formation on tyrosyl peptides lacking a terminal amine, without forming an adduct. These findings, plus the high hydroperoxide yields with N-terminal tyrosine, can be explained by a mechanism in which hydrogen bonding of O(2)(.) to the amine increases is oxidizing potential and alters its reactivity. If this amine effect occurred more generally, it could increase the biological reactivity of O(2)(.) and have major implications. PMID- 19297320 TI - Adenosine signaling mediates SUMO-1 modification of IkappaBalpha during hypoxia and reoxygenation. AB - Small ubiquitin-like modifier 1 (SUMO-1) modification of IkappaBalpha has been described to actively participate in NFkappaB regulation. Following proteosomal degradation of IkappaBalpha, an auto-regulatory loop consisting of transcriptional activation of IkappaBalpha gene and SUMO-1 modification of newly synthesized IkappaBalpha proceeds. The SUMOylated IkappaBalpha form is resistant to signal-induced degradation, consequently halting NFkappaB activation. We describe a mechanistic model by which adenosine (Ado) signaling results in significant accumulation of SUMO-1 modified IkappaBalpha with subsequent attenuation of NFkappaB activation. Using models of hypoxia followed by reoxygenation (H/R), we have documented an H/R cycle-dependent increase in extracellular Ado correlating with increases in the cytoplasmic pool of IkappaBalpha/SUMO-1. We demonstrate a dose-dependent increase in IkappaBalpha/SUMO in cells treated with the general Ado receptor agonist NECA and abolished by Ado receptor antagonists. Experiments in cells exposed to cycles of H/R followed by hypoxia demonstrated differential patterns of SUMOylation and phosphorylation of IkappaBalpha, greatly impacting its proteosomal degradation by the 26 S proteasome. Assays targeting knockdown and overexpression of SUMO-1 demonstrated significant regulation of NFkappaB activation and NFkappaB-mediated gene transcription (interleukin-6). These results were confirmed in vivo using wild type and cd73 null mouse lung tissue. In summary, we present an endogenous mechanism by which cells and tissues acquire anti-inflammatory properties by recruiting a nondegradable form of IkappaBalpha, a major control point for NFkappaB activation via Ado signaling. PMID- 19297321 TI - Hepatitis C virus NS5A protein is a substrate for the peptidyl-prolyl cis/trans isomerase activity of cyclophilins A and B. AB - We report here a biochemical and structural characterization of domain 2 of the nonstructural 5A protein (NS5A) from the JFH1 Hepatitis C virus strain and its interactions with cyclophilins A and B (CypA and CypB). Gel filtration chromatography, circular dichroism spectroscopy, and finally NMR spectroscopy all indicate the natively unfolded nature of this NS5A-D2 domain. Because mutations in this domain have been linked to cyclosporin A resistance, we used NMR spectroscopy to investigate potential interactions between NS5A-D2 and cellular CypA and CypB. We observed a direct molecular interaction between NS5A-D2 and both cyclophilins. The interaction surface on the cyclophilins corresponds to their active site, whereas on NS5A-D2, it proved to be distributed over the many proline residues of the domain. NMR heteronuclear exchange spectroscopy yielded direct evidence that many proline residues in NS5A-D2 form a valid substrate for the enzymatic peptidyl-prolyl cis/trans isomerase (PPIase) activity of CypA and CypB. PMID- 19297322 TI - Four-electron reduction of dioxygen by a multicopper oxidase, CueO, and roles of Asp112 and Glu506 located adjacent to the trinuclear copper center. AB - The mechanism of the four-electron reduction of dioxygen by a multicopper oxidase, CueO, was studied based on reactions of single and double mutants with Cys(500), a type I copper ligand, and the noncoordinating Asp(112) and Glu(506), which form hydrogen bonds with the trinuclear copper center directly and indirectly via a water molecule. The reaction of C500S containing a vacant type I copper center produced intermediate I in an EPR-silent peroxide-bound form. The formation of intermediate I from C500S/D112N was restricted due to a reduction in the affinity of the trinuclear copper center for dioxygen. The state of intermediate I was realized to be the resting form of C500S/E506Q and C500S of the truncated mutant Deltaalpha5-7CueO, in which the 50 amino acids covering the substrate-binding site were removed. Reactions of the recombinant CueO and E506Q afforded intermediate II, a fully oxidized form different from the resting one, with a very broad EPR signal, g < 2, detectable only at cryogenic temperatures and unsaturated with high power microwaves. The lifetime of intermediate II was prolonged by the mutation at Glu(506) involved in the donation of protons. The structure of intermediates I and II and the mechanism of the four-electron reduction of dioxygen driven by Asp(112) and Glu(506) are discussed. PMID- 19297323 TI - CRM1-mediated nuclear export of dengue virus RNA polymerase NS5 modulates interleukin-8 induction and virus production. AB - Although all established functions of dengue virus NS5 (nonstructural protein 5) occur in the cytoplasm, its nuclear localization, mediated by dual nuclear localization sequences, is essential for virus replication. Here, we have determined the mechanism by which NS5 can localize in the cytoplasm to perform its role in replication, establishing for the first time that it is able to be exported from the nucleus by the exportin CRM1 and hence can shuttle between the nucleus and cytoplasm. We define the nuclear export sequence responsible to be residues 327-343 and confirm interaction of NS5 and CRM1 by pulldown assay. Significantly, greater nuclear accumulation of NS5 during infection due to CRM1 inhibition coincided with altered kinetics of virus production and decreased induction of the antiviral chemokine interleukin-8. This is the first report of a nuclear export sequence within NS5 for any member of the Flavivirus genus; because of its high conservation within the genus, it may represent a target for the treatment of diseases caused by several medically important flaviviruses. PMID- 19297324 TI - Structural insight into the activation mechanism of human pancreatic prophospholipase A2. AB - Pancreatic phospholipase A2 (phospholipase A2 group 1B, G1B) belongs to the superfamily of secreted phospholipase A2 (PLA2) enzymes. G1B has been proposed to be a potential target for diseases such as hypertension, obesity, and diabetes. Human pancreatic prophospholipase A2 (pro-hG1B) is activated by cleavage of the first seven-residue propeptide (phospholipase A2 propeptide, PROP). However, questions still remain on the mode of action for pro-hG1B. In this work, we expressed pro-hG1B in Pichia pastoris and determined the crystal structure at 1.55-A resolution. The x-ray structure demonstrates that pro-hG1B forms a trimer. In addition, PROP occupies the catalytic cavity and can be self-cleaved at 37 degrees C. A new membrane-bound surface and activation mechanism are proposed based on the trimeric model of pro-hG1B. We also propose a new autoproteolytic mechanism for pro-hG1B by the reaction triad Asp49-Arg0-Ser(-2) that is similar to the serine protease catalytic triad. PMID- 19297325 TI - LFA-1 affinity regulation is necessary for the activation and proliferation of naive T cells. AB - The activation of LFA-1 (lymphocyte function-associated antigen) is a critical event for T cell co-stimulation. The mechanism of LFA-1 activation involves both affinity and avidity regulation, but the role of each in T cell activation remains unclear. We have identified antibodies that recognize and block different affinity states of the mouse LFA-1 I-domain. Monoclonal antibody 2D7 preferentially binds to the low affinity conformation, and this specific binding is abolished when LFA-1 is locked in the high affinity conformation. In contrast, M17/4 can bind both the locked high and low affinity forms of LFA-1. Although both 2D7 and M17/4 are blocking antibodies, 2D7 is significantly less potent than M17/4 in blocking LFA-1-mediated adhesion; thus, blocking high affinity LFA-1 is critical for preventing LFA-1-mediated adhesion. Using these reagents, we investigated whether LFA-1 affinity regulation affects T cell activation. We found that blocking high affinity LFA-1 prevents interleukin-2 production and T cell proliferation, demonstrated by TCR cross-linking and antigen-specific stimulation. Furthermore, there is a differential requirement of high affinity LFA-1 in the activation of CD4(+) and CD8(+) T cells. Although CD4(+) T cell activation depends on both high and low affinity LFA-1, only high affinity LFA-1 provides co-stimulation for CD8(+) T cell activation. Together, our data demonstrated that the I-domain of LFA-1 changes to the high affinity state in primary T cells, and high affinity LFA-1 is critical for facilitating T cell activation. This implicates LFA-1 activation as a novel regulatory mechanism for the modulation of T cell activation and proliferation. PMID- 19297326 TI - Inhibition of tumor cell motility by the interferon-inducible GTPase MxA. AB - To identify pathways controlling prostate cancer metastasis we performed differential display analysis of the human prostate carcinoma cell line PC-3 and its highly metastatic derivative PC-3M. This revealed that a 78-kDa interferon inducible GTPase, MxA, was expressed in PC-3 but not in PC-3M cells. The gene encoding MxA, MX1, is located in the region of chromosome 21 deleted as a consequence of fusion of TMPRSS2 and ERG, which has been associated with aggressive, invasive prostate cancer. Stable exogenous MxA expression inhibited in vitro motility and invasiveness of PC-3M cells. In vivo exogenous MxA expression decreased the number of hepatic metastases following intrasplenic injection. Exogenous MxA also reduced motility and invasiveness of highly metastatic LOX melanoma cells. A mutation in MxA that inactivated its GTPase reversed inhibition of motility and invasion in both tumor cell lines. Co immunoprecipitation studies demonstrated that MxA associated with tubulin, but the GTPase-inactivating mutation blocked this association. Because MxA is a highly inducible gene, an MxA-targeted drug discovery screen was initiated by placing the MxA promoter upstream of a luciferase reporter. Examination of the NCI diversity set of small molecules revealed three hits that activated the promoter. In PC-3M cells, these drugs induced MxA protein and inhibited motility. These data demonstrate that MxA inhibits tumor cell motility and invasion, and that MxA expression can be induced by small molecules, potentially offering a new approach to the prevention and treatment of metastasis. PMID- 19297327 TI - Active plasma kallikrein localizes to mast cells and regulates epithelial cell apoptosis, adipocyte differentiation, and stromal remodeling during mammary gland involution. AB - The plasminogen cascade of serine proteases directs both development and tumorigenesis in the mammary gland. Plasminogen can be activated to plasmin by urokinase-type plasminogen activator (uPA), tissue-type plasminogen activator (tPA), and plasma kallikrein (PKal). The dominant plasminogen activator for mammary involution is PKal, a serine protease that participates in the contact activation system of blood coagulation. We observed that the prekallikrein gene (Klkb1) is expressed highly in the mammary gland during stromal remodeling periods including puberty and postlactational involution. We used a variant of ecotin (ecotin-PKal), a macromolecular inhibitor of serine proteases engineered to be highly specific for active PKal, to demonstrate that inhibition of PKal with ecotin-PKal delays alveolar apoptosis, adipocyte replenishment, and stromal remodeling in the involuting mammary gland, producing a phenotype resembling that resulting from plasminogen deficiency. Using biotinylated ecotin-PKal, we localized active PKal to connective tissue-type mast cells in the mammary gland. Taken together, these results implicate PKal as an effector of the plasminogen cascade during mammary development. PMID- 19297328 TI - The F-box protein beta-TrCp1/Fbw1a interacts with p300 to enhance beta-catenin transcriptional activity. AB - Hyperactivated beta-catenin is a commonly found molecular abnormality in colon cancer, and its nuclear accumulation is thought to promote the expression of genes associated with cellular proliferation and transformation. The p300 transcriptional co-activator binds to beta-catenin and facilitates transcription by recruiting chromatin remodeling complexes and general transcriptional apparatus. We have found that beta-TrCp1/Fbw1a, a member of the Skp1/Cullin/Rbx1/F-box E3 ubiquitin ligase complex, binds directly to p300 and co localizes with it to beta-catenin target gene promoters. Our data show that Fbw1a, which normally targets beta-catenin for degradation, works together with p300 to enhance the transcriptional activity of beta-catenin, whereas other F box/WD40 proteins do not. Fbw1a also cooperates with p300 to co-activate transcription by SMAD3, another Fbw1a ubiquitylation target, but not p53 or HIF 1alpha, which are substrates for other ubiquitin ligase complexes. These results suggest that, although Fbw1a is part of a negative feedback loop for controlling beta-catenin levels in normal cells, its overexpression and binding to p300 may contribute to hyperactivated beta-catenin transcriptional activity in colon cancer cells. PMID- 19297329 TI - Stereospecificity of the siderophore pyochelin outer membrane transporters in fluorescent pseudomonads. AB - Pyochelin (Pch) and enantio-pyochelin (EPch) are enantiomer siderophores that are produced by Pseudomonas aeruginosa and Pseudomonas fluorescens, respectively, under iron limitation. Pch promotes growth of P. aeruginosa when iron is scarce, and EPch carries out the same biological function in P. fluorescens. However, the two siderophores are unable to promote growth in the heterologous species, indicating that siderophore-mediated iron uptake is highly stereospecific. In the present work, using binding and iron uptake assays, we found that FptA, the Fe Pch outer membrane transporter of P. aeruginosa, recognized (K(d) = 2.5 +/- 1.1 nm) and transported Fe-Pch but did not interact with Fe-EPch. Likewise, FetA, the Fe-EPch receptor of P. fluorescens, was specific for Fe-EPch (K(d) = 3.7 +/- 2.1 nm) but did not bind and transport Fe-Pch. Growth promotion experiments performed under iron-limiting conditions confirmed that FptA and FetA are highly specific for Pch and EPch, respectively. When fptA and fetA along with adjacent transport genes involved in siderophore uptake were swapped between the two bacterial species, P. aeruginosa became able to utilize Fe-EPch as an iron source, and P. fluorescens was able to grow with Fe-Pch. Docking experiments using the FptA structure and binding assays showed that the stereospecificity of Pch recognition by FptA was mostly due to the configuration of the siderophore chiral centers C4'' and C2'' and was only weakly dependent on the configuration of the C4' carbon atom. Together, these findings increase our understanding of the stereospecific interaction between Pch and its outer membrane receptor FptA. PMID- 19297330 TI - Promiscuous usage of nucleotides by the DNA helicase of bacteriophage T7: determinants of nucleotide specificity. AB - The multifunctional protein encoded by gene 4 of bacteriophage T7 (gp4) provides both helicase and primase activity at the replication fork. T7 DNA helicase preferentially utilizes dTTP to unwind duplex DNA in vitro but also hydrolyzes other nucleotides, some of which do not support helicase activity. Very little is known regarding the architecture of the nucleotide binding site in determining nucleotide specificity. Crystal structures of the T7 helicase domain with bound dATP or dTTP identified Arg-363 and Arg-504 as potential determinants of the specificity for dATP and dTTP. Arg-363 is in close proximity to the sugar of the bound dATP, whereas Arg-504 makes a hydrogen bridge with the base of bound dTTP. T7 helicase has a serine at position 319, whereas bacterial helicases that use rATP have a threonine in the comparable position. Therefore, in the present study we have examined the role of these residues (Arg-363, Arg-504, and Ser-319) in determining nucleotide specificity. Our results show that Arg-363 is responsible for dATP, dCTP, and dGTP hydrolysis, whereas Arg-504 and Ser-319 confer dTTP specificity. Helicase-R504A hydrolyzes dCTP far better than wild-type helicase, and the hydrolysis of dCTP fuels unwinding of DNA. Substitution of threonine for serine 319 reduces the rate of hydrolysis of dTTP without affecting the rate of dATP hydrolysis. We propose that different nucleotides bind to the nucleotide binding site of T7 helicase by an induced fit mechanism. We also present evidence that T7 helicase uses the energy derived from the hydrolysis of dATP in addition to dTTP for mediating DNA unwinding. PMID- 19297331 TI - Systems biological analysis of epidermal growth factor receptor internalization dynamics for altered receptor levels. AB - Epidermal growth factor (EGF) receptor (EGFR) overexpression is a hallmark of many cancers. EGFR endocytosis is a critical step in signal attenuation, raising the question of how receptor expression levels affect the internalization process. Here we combined quantitative experimental and mathematical modeling approaches to investigate the role of the EGFR expression level on the rate of receptor internalization. Using tetramethylrhodamine-labeled EGF, we established assays for quantifying EGF-triggered EGFR internalization by both high resolution confocal microscopy and flow cytometry. We determined that the flow cytometry approach was more sensitive for examining large populations of cells. Mathematical modeling was used to investigate the relationship between EGF internalization kinetics, EGFR expression, and internalization machinery. We predicted that the standard parameter used to assess internalization kinetics, the temporal evolution r(t) of the ratio of internalized versus surface-located ligand.receptor complexes, does not describe a straight line, as proposed previously. Instead, a convex or concave curve occurs depending on whether initial receptor numbers or internalization adaptors are limiting the uptake reaction, respectively. To test model predictions, we measured EGF-EGFR binding and internalization in cells expressing different levels of green fluorescent protein-EGFR. As expected, surface binding of rhodamine-labeled EGF increased with green fluorescent protein-EGFR expression level. Unexpectedly, internalization of ligand. receptor complexes increased linearly with increasing receptor expression level, suggesting that receptors and not internalization adaptors were limiting the uptake in our experimental model. Finally, determining the ratio of internalized versus surface-located ligand.receptor complexes for this cell line confirmed that it follows a convex curve, supporting our model predictions. PMID- 19297333 TI - FoxO1 controls insulin-dependent adipose triglyceride lipase (ATGL) expression and lipolysis in adipocytes. AB - FoxO1 represents a central regulator of metabolism in several cell types. Although FoxO1 is abundant in adipocytes, its biological functions in these cells remain largely unknown. We show here that the promotor region of the rate limiting lipolytic enzyme, adipose triglyceride lipase (ATGL), has two FoxO1 binding sites, and co-transfection with wild type and unphosphorylated FoxO1 mutant activates the expression of luciferase driven by the ATGL promotor. In 3T3 L1 adipocytes, insulin controls nucleo-cytoplasmic shuttling of FoxO1 and regulates its interaction with endogenous ATGL promotors. Knockdown of FoxO1 in 3T3-L1 adipocytes decreases the expression of ATGL and attenuates basal and isoproterenol-stimulated lipolysis. Infection of mouse embryonic fibroblasts with FoxO1-encoding lentivirus increases ATGL expression and renders it sensitive to regulation by insulin. Thus, FoxO1 may play an important role in the regulation of lipolysis in adipocytes by controlling the expression of ATGL. PMID- 19297332 TI - Adenine nucleotide-dependent regulation of assembly of bacterial tubulin-like FtsZ by a hypermorph of bacterial actin-like FtsA. AB - Cytokinesis in bacteria depends upon the contractile Z ring, which is composed of dynamic polymers of the tubulin homolog FtsZ as well as other membrane-associated proteins such as FtsA, a homolog of actin that is required for membrane attachment of the Z ring and its subsequent constriction. Here we show that a previously characterized hypermorphic mutant FtsA (FtsA*) partially disassembled FtsZ polymers in vitro. This effect was strictly dependent on ATP or ADP binding to FtsA* and occurred at substoichiometric levels relative to FtsZ, similar to cellular levels. Nucleotide-bound FtsA* did not affect FtsZ GTPase activity or the critical concentration for FtsZ assembly but was able to disassemble preformed FtsZ polymers, suggesting that FtsA* acts on FtsZ polymers. Microscopic examination of the inhibited FtsZ polymers revealed a transition from long, straight polymers and polymer bundles to mainly short, curved protofilaments. These results indicate that a bacterial actin, when activated by adenine nucleotides, can modify the length distribution of bacterial tubulin polymers, analogous to the effects of actin-depolymerizing factor/cofilin on F-actin. PMID- 19297334 TI - The structure of an interdomain complex that regulates talin activity. AB - Talin is a large flexible rod-shaped protein that activates the integrin family of cell adhesion molecules and couples them to cytoskeletal actin. It exists in both globular and extended conformations, and an intramolecular interaction between the N-terminal F3 FERM subdomain and the C-terminal part of the talin rod contributes to an autoinhibited form of the molecule. Here, we report the solution structure of the primary F3 binding domain within the C-terminal region of the talin rod and use intermolecular nuclear Overhauser effects to determine the structure of the complex. The rod domain (residues 1655-1822) is an amphipathic five-helix bundle; Tyr-377 of F3 docks into a hydrophobic pocket at one end of the bundle, whereas a basic loop in F3 (residues 316-326) interacts with a cluster of acidic residues in the middle of helix 4. Mutation of Glu-1770 abolishes binding. The rod domain competes with beta3-integrin tails for binding to F3, and the structure of the complex suggests that the rod is also likely to sterically inhibit binding of the FERM domain to the membrane. PMID- 19297335 TI - Full domain closure of the ligand-binding core of the ionotropic glutamate receptor iGluR5 induced by the high affinity agonist dysiherbaine and the functional antagonist 8,9-dideoxyneodysiherbaine. AB - The prevailing structural model for ligand activation of ionotropic glutamate receptors posits that agonist efficacy arises from the stability and magnitude of induced domain closure in the ligand-binding core structure. Here we describe an exception to the correlation between ligand efficacy and domain closure. A weakly efficacious partial agonist of very low potency for homomeric iGluR5 kainate receptors, 8,9-dideoxyneodysiherbaine (MSVIII-19), induced a fully closed iGluR5 ligand-binding core. The degree of relative domain closure, approximately 30 degrees , was similar to that we resolved with the structurally related high affinity agonist dysiherbaine and to that of l-glutamate. The pharmacological activity of MSVIII-19 was confirmed in patch clamp recordings from transfected HEK293 cells, where MSVIII-19 predominantly inhibits iGluR5-2a, with little activation apparent at a high concentration (1 mm) of MSVIII-19 (<1% of mean glutamate-evoked currents). To determine the efficacy of the ligand quantitatively, we constructed concentration-response relationships for MSVIII-19 following potentiation of steady-state currents with concanavalin A (EC(50) = 3.6 microm) and on the nondesensitizing receptor mutant iGluR5-2b(Y506C/L768C) (EC(50) = 8.1 microm). MSVIII-19 exhibited a maximum of 16% of full agonist efficacy, as measured in parallel recordings with glutamate. Molecular dynamics simulations and electrophysiological recordings confirm that the specificity of MSVIII-19 for iGluR5 is partly attributable to interdomain hydrogen bond residues Glu(441) and Ser(721) in the iGluR5-S1S2 structure. The weaker interactions of MSVIII-19 with iGluR5 compared with dysiherbaine, together with altered stability of the interdomain interaction, may be responsible for the apparent uncoupling of domain closure and channel opening in this kainate receptor subunit. PMID- 19297336 TI - Smoking, BMI and psychological strain and fitness in the Naval Service. AB - BACKGROUND: Data from the Naval Service (NS) cohort study of psychological strain were extracted and analysed to investigate the relationship between self-reported health and lifestyle factors and medical fitness. Identification of factors associated with medical downgrading is of obvious value in shaping future health and safety policy and in understanding the relative contributions of physical and psychosocial factors to adverse occupational health outcomes. AIMS: To identify variables associated with a lack of fitness to serve. METHOD: Extraction and analysis of data from the Phase I of the study, with a binary outcome of fitness as the dependent variable, controlling for psychosocial and other confounders. RESULTS: Stepwise logistic regression analysis found statistically significant effects due to smoking, body mass index (BMI), General Health Questionnaire (GHQ) 12 and work-family conflict. The model accounted for 5.6% of the variance in medical grading, 3% of which was due to smoking. CONCLUSIONS: With psychosocial factors and GHQ-12 scores accounted for, personnel who were not fully fit for NS were found to be more likely to be smokers and to have a high BMI (> or =25). PMID- 19297337 TI - Mortality in employees at a New Zealand agrochemical manufacturing site. AB - BACKGROUND: Previous studies at the Dow AgroSciences (Formerly Ivon Watkins-Dow) plant in New Plymouth, New Zealand, had raised concerns about the cancer risk in a subset of workers at the site with potential exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin. As the plant had been involved in the synthesis and formulation of a wide range of agrochemicals and their feedstocks, we examined the mortality risk for all workers at the site. AIMS: To quantify the mortality hazards arising from employment at the Dow AgroSciences agrochemical production site in New Plymouth, New Zealand. METHODS: Workers employed between 1 January 1969 and 1 October 2003 were followed up to the end of 2004. Standardized mortality ratios (SMRs) were calculated using national mortality rates by employment duration, sex, period of hire and latency. RESULTS: A total of 1754 employees were followed during the study period and 247 deaths were observed. The all causes and all cancers SMRs were 0.97 (95% CI 0.85-1.10) and 1.01 (95% CI 0.80-1.27), respectively. Mortality due to all causes was higher for short-term workers (SMR 1.23, 95% CI 0.91-1.62) than long-term workers (SMR 0.92, 95% CI 0.80-1.06) and women had lower death rates than men. Analyses by latency and period of hire did not show any patterns consistent with an adverse impact of occupational exposures. CONCLUSIONS: The mortality experience of workers at the site was similar to the rest of New Zealand. PMID- 19297338 TI - Occupational dermatoses in restaurant, catering and fast-food outlets in Singapore. AB - BACKGROUND: The restaurant industry is a rapidly growing sector in Singapore and workers in this industry are trained in culinary skills but not on recognition of safety and health hazards and their control measures. Anecdotal clinical evidence has suggested an increased prevalence of occupational dermatoses among restaurant workers. AIMS: To determine the prevalence and risk factors for contact dermatitis and burns among restaurant, catering and fast-food outlet (FFO) staff. METHODS: Workers were interviewed and then clinical examination and patch and/or prick tests were conducted in selected individuals. RESULTS: In total, 335 of 457 workers (73% response) were interviewed and 65 (19%) had occupational dermatitis or burns and were examined. Of these, contact dermatitis was the commonest diagnosis, with a 12-month period prevalence of 10% (35 workers) and 3-month period prevalence of 8% (26 workers). All 35 workers had irritant contact dermatitis (ICD) and there were no cases of allergic contact dermatitis. The adjusted prevalence rate ratios of risk factors for ICD were 2.78 (95% CI 1.36 5.72) for frequent hand washing >20 times per day, 3.87 (95% CI 1.89-7.93) for atopy and 2.57 (95% CI 1.21-5.47) for contact with squid. The 3-month period prevalence for burns was 6% (20 workers). Ten workers had other occupational dermatoses such as work-related calluses, paronychia, heat rash and allergic contact urticaria to prawn and lobster. CONCLUSIONS: ICD and burns are common occupational skin disorders among restaurant, catering and FFO workers. PMID- 19297339 TI - Inside the fitness for work consultation: a qualitative study. AB - BACKGROUND: Evidence now suggests that work is generally good for physical and mental health and well-being. Worklessness for whatever reason can lead to poorer physical and mental health. The role of the general practitioner (GP) in the management of fitness for work is pivotal. AIMS: To understand the interaction between GP and patient in the fitness for work consultation. This study forms part of a larger research project to develop a learning programme for GPs around the fitness for work consultation based on behaviour change methodology. METHODS: A qualitative study set in South Wales. Structured discussion groups with seven GPs. Two sessions each lasting 3 h were conducted to explore the GP and patient interaction around the fitness for work consultation. Multiple methods were used to enhance engagement. Thematic analysis was used to analyse the data. RESULTS: Four major themes emerged from the meetings: role legitimacy, negotiation, managing the patient and managing the systems. Within these, subthemes emerged around role legitimacy. 'It's not my job', 'It's not what I trained for' and the 'shifting agenda' Negotiation was likened to 'A polite tug of war' and subthemes around decision making, managing the agenda and dealing with uncertainty emerged. CONCLUSIONS: This study starts to unravel the complexity of the fitness for work consultation. It illustrates how GPs struggle with the 'importance' of their role and 'confidence' in managing the fitness for work consultation. It addresses the skillful negotiation that is required to manage the consultation effectively. PMID- 19297340 TI - Use of Doppler in the diagnosis of hypothenar hammer syndrome. AB - BACKGROUND: Hand-arm vibration syndrome (HAVS) includes a spectrum of vascular, neurological and musculoskeletal symptoms resulting from exposure to vibrating tools. Hypothenar hammer syndrome (HHS) is a lesion of the ulnar artery as it courses adjacent to the hamate bone and results from either single or repeated episodes of trauma to the hypothenar eminence. There is a need to distinguish symptoms of HHS from those of classical HAVS since precise diagnosis may alter both the clinical and occupational management of the affected employee. AIMS: To highlight the value of simple Doppler assessments of the palmar blood flow to distinguish the condition of HHS from 'classical' HAVS. METHOD: Among patients assessed for HAVS by the authors during 2006, three were identified as potentially having HHS. Doppler ultrasound of the palmar arches with and without ulnar arterial occlusion was used. RESULT: We report three cases in which Doppler ultrasound assessment supports a diagnosis of HHS. CONCLUSIONS: It is our recommendation that such Doppler assessments should form part of the clinical assessment of workers being assessed in connection with exposure to hand transmitted vibration and in whom symptoms are present that are not typical of 'classical HAVS', particularly where there is a history of possible hypothenar trauma. PMID- 19297341 TI - The relationship of body mass index and blood pressure in Iranian children <7 years old. AB - The objective of this study was to evaluate the association between the body mass index (BMI), in healthy young children with their blood pressure (BP). The study included 3186 healthy children aged 1-6 years who were studied between March 2004 and March 2007 in different kindergartens and health centers in Tehran. Each child was classified on the basis of age- and sex-specific BMI percentile as normal weight (BMI <85th percentile), at risk for overweight (BMI >85th and <95th percentile), or overweight (BMI > or =95th percentile). Systolic BP (SBP) and diastolic BP (DBP) was compared among age-sex-BMI groups. Among children aged below 7 years in kindergartens and health centers in Tehran, 7.2% were at risk of overweight and 12.2% were overweight. These proportions were similar for boys and girls and were as follows: 6.9%, 13.9% and 7.5%, 10.5%, respectively. Analysis of variance showed that mean SBP significantly increased according to age (p < 0.0001) and BMI group (p = 0.001). Analysis of variance also showed that mean DBP significantly increased as age increased (p < 0.0001), but no significant difference was found between boys and girls in different age and BMI groups (p = 0.37). Our survey identified a high prevalence of overweight that was associated with elevated SBP among preschool-aged children in Iran. The effect of higher BMI on mean SBP is present in childhood and can be used as a predictor of high SBP even in children as young as 1-6 years. PMID- 19297342 TI - A study of profile of ventilator-associated pneumonia in children in Punjab. AB - OBJECTIVES: To determine incidence, related factors, outcome, bacterial organisms and their sensitivity patterns with regard to ventilator-associated pneumonia (VAP) in children. SETTING: Level III PICU of a tertiary care center. DESIGN: Prospective cohort study. METHODS: Children in the age group of 1 month to 15 years, admitted to the pediatric intensive care unit requiring ventilatory support (V.I.P.BIRD infant-Pediatric ventilator) for at least 48 h. Clinical criteria used to define VAPs were the same as used by and Elward et al. and Salata et al. RESULTS: Forty patients met the inclusion criteria and 8 (20%) had VAP. The risk factor significantly related with development of VAP was the use of H(2) blockers (Ranitidine) for >2 days. All other related factors were not significantly related to occurrence of VAP. CONCLUSION: Use of H(2) blockers (Ranitidne) is associated with higher incidence of VAP in children. PMID- 19297343 TI - Application of two-dimensional crystallography and image processing to atomic resolution Z-contrast images. AB - Zone axis images recorded using high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM or Z-contrast imaging) reveal the atomic structure with a resolution that is defined by the probe size of the microscope. In most cases, the full images contain many sub-images of the crystal unit cell and/or interface structure. Thanks to the repetitive nature of these images, it is possible to apply standard image processing techniques that have been developed for the electron crystallography of biological macromolecules and have been used widely in other fields of electron microscopy for both organic and inorganic materials. These methods can be used to enhance the signal-to-noise present in the original images, to remove distortions in the images that arise from either the instrumentation or the specimen itself and to quantify properties of the material in ways that are difficult without such data processing. In this paper, we describe briefly the theory behind these image processing techniques and demonstrate them for aberration-corrected, high-resolution HAADF-STEM images of Si(46) clathrates developed for hydrogen storage. PMID- 19297344 TI - The development of Assessment of SpondyloArthritis international Society classification criteria for axial spondyloarthritis (part II): validation and final selection. AB - OBJECTIVE: To validate and refine two sets of candidate criteria for the classification/diagnosis of axial spondyloarthritis (SpA). METHODS: All Assessment of SpondyloArthritis international Society (ASAS) members were invited to include consecutively new patients with chronic (> or =3 months) back pain of unknown origin that began before 45 years of age. The candidate criteria were first tested in the entire cohort of 649 patients from 25 centres, and then refined in a random selection of 40% of cases and thereafter validated in the remaining 60%. RESULTS: Upon diagnostic work-up, axial SpA was diagnosed in 60.2% of the cohort. Of these, 70% did not fulfil modified New York criteria and, therefore, were classified as having "non-radiographic" axial SpA. Refinement of the candidate criteria resulted in new ASAS classification criteria that are defined as: the presence of sacroiliitis by radiography or by magnetic resonance imaging (MRI) plus at least one SpA feature ("imaging arm") or the presence of HLA-B27 plus at least two SpA features ("clinical arm"). The sensitivity and specificity of the entire set of the new criteria were 82.9% and 84.4%, and for the imaging arm alone 66.2% and 97.3%, respectively. The specificity of the new criteria was much better than that of the European Spondylarthropathy Study Group criteria modified for MRI (sensitivity 85.1%, specificity 65.1%) and slightly better than that of the modified Amor criteria (sensitivity 82.9, specificity 77.5%). CONCLUSION: The new ASAS classification criteria for axial SpA can reliably classify patients for clinical studies and may help rheumatologists in clinical practice in diagnosing axial SpA in those with chronic back pain. TRIAL REGISTRATION NUMBER: NCT00328068. PMID- 19297345 TI - The development of Assessment of SpondyloArthritis international Society classification criteria for axial spondyloarthritis (part I): classification of paper patients by expert opinion including uncertainty appraisal. AB - OBJECTIVE: Non-radiographic axial spondyloarthritis (SpA) is characterised by a lack of definitive radiographic sacroiliitis and is considered an early stage of ankylosing spondylitis. The objective of this study was to develop candidate classification criteria for axial SpA that include patients with but also without radiographic sacroiliitis. METHODS: Seventy-one patients with possible axial SpA, most of whom were lacking definite radiographic sacroiliitis, were reviewed as "paper patients" by 20 experts from the Assessment of SpondyloArthritis international Society (ASAS). Unequivocally classifiable patients were identified based on the aggregate expert opinion in conjunction with the expert-reported level of certainty of their judgement. Draft criteria for axial SpA were formulated and tested using classifiable patients. RESULTS: Active sacroiliitis on magnetic resonance imaging (MRI) (odds ratio 45, 95% CI 5.3 to 383; p<0.001) was strongly associated with the classification of axial SpA. The knowledge of MRI findings led to a change in the classification of 21.1% of patients. According to the first set of candidate criteria (sensitivity 97.1%; specificity 94.7%) a patient with chronic back pain is classified as axial SpA in the presence of sacroiliitis by MRI or x rays in conjunction with one SpA feature or, if sacroilitiis is absent, in the presence of at least three SpA features. In a second set of candidate criteria, inflammatory back pain is obligatory in the clinical arm (sensitivity 86.1%; specificity 94.7%). CONCLUSION: The ASAS group has developed candidate criteria for the classification of axial SpA that include patients without radiographic sacroiliitis. The candidate criteria need to be validated in an independent international study. PMID- 19297346 TI - Comparison of tocilizumab monotherapy versus methotrexate monotherapy in patients with moderate to severe rheumatoid arthritis: the AMBITION study. AB - BACKGROUND: The anti-interleukin (IL) 6 receptor antibody tocilizumab inhibits signalling of IL6, a key cytokine in rheumatoid arthritis (RA) pathogenesis. OBJECTIVE: To evaluate through the AMBITION study the efficacy and safety of tocilizumab monotherapy versus methotrexate in patients with active RA for whom previous treatment with methotrexate/biological agents had not failed. METHODS: This 24-week, double-blind, double-dummy, parallel-group study, randomised 673 patients to either tocilizumab 8 mg/kg every 4 weeks, or methotrexate, starting at 7.5 mg/week and titrated to 20 mg/week within 8 weeks, or placebo for 8 weeks followed by tocilizumab 8 mg/kg. The primary end point was the proportion of patients achieving American College of Rheumatology (ACR) 20 response at week 24. RESULTS: The intention-to-treat analysis demonstrated that tocilizumab was better than methotrexate treatment with a higher ACR20 response (69.9 vs 52.5%; p<0.001), and 28-joint Disease Activity Score (DAS28) <2.6 rate (33.6 vs 12.1%) at week 24. Mean high-sensitivity C-reactive protein was within the normal range from week 12 with tocilizumab, whereas levels remained elevated with methotrexate. The incidence of serious adverse events with tocilizumab was 3.8% versus 2.8% with methotrexate (p = 0.50), and of serious infections, 1.4% versus 0.7%, respectively. There was a higher incidence of reversible grade 3 neutropenia (3.1% vs 0.4%) and increased total cholesterol > or =240 mg/dl (13.2% vs 0.4%), and a lower incidence of alanine aminotransferase elevations >3x-<5x upper limit of normal (1.0% vs 2.5%), respectively. CONCLUSION: Tocilizumab monotherapy is better than methotrexate monotherapy, with rapid improvement in RA signs and symptoms, and a favourable benefit-risk, in patients for whom treatment with methotrexate or biological agents has not previously failed. PMID- 19297347 TI - Synergy Disequilibrium Plots: graphical visualization of pairwise synergies and redundancies of SNPs with respect to a phenotype. AB - SUMMARY: We present a visualization tool applied on genome-wide association data, revealing disease-associated haplotypes, epistatically interacting loci, as well as providing visual signatures of multivariate correlations of genetic markers with respect to a phenotype. AVAILABILITY: Freely available on the web at: (http://www.ee.columbia.edu/~anastas/sdplots). PMID- 19297348 TI - Visualization of genomic data with the Hilbert curve. AB - In many genomic studies, one works with genome-position-dependent data, e.g. ChIP chip or ChIP-Seq scores. Using conventional tools, it can be difficult to get a good feel for the data, especially the distribution of features. This article argues that the so-called Hilbert curve visualization can complement genome browsers and help to get further insights into the structure of one's data. This is demonstrated with examples from different use cases. An open-source application, called HilbertVis, is presented that allows the user to produce and interactively explore such plots. AVAILABILITY: http://www.ebi.ac.uk/huber srv/hilbert/. PMID- 19297349 TI - DASher: a stand-alone protein sequence client for DAS, the Distributed Annotation System. AB - SUMMARY: The rise in biological sequence data has led to a proliferation of separate, specialized databases. While there is great value in having many independent annotations, it is critical that there be a way to integrate them in one combined view. The Distributed Annotation System (DAS) was developed for that very purpose. There are currently no DAS clients that are open source, specialized for aggregating and comparing protein sequence annotation, and that can run as a self-contained application outside of a web browser. The speed, flexibility and extensibility that come with a stand-alone application motivated us to create DASher, an open-source Java DAS client. Given a UniProt sequence identifier, DASher automatically queries DAS-supporting servers worldwide for any information on that sequence and then displays the annotations in an interactive viewer for easy comparison. DASher is a fast, Java-based DAS client optimized for viewing protein sequence annotation and compliant with the latest DAS protocol specification 1.53E. AVAILABILITY: DASher is available for direct use and download at http://dasher.sbc.su.se including examples and source code under the GPLv3 licence. Java version 6 or higher is required. PMID- 19297350 TI - Artefacts and biases affecting the evaluation of scoring functions on decoy sets for protein structure prediction. AB - MOTIVATION: Decoy datasets, consisting of a solved protein structure and numerous alternative native-like structures, are in common use for the evaluation of scoring functions in protein structure prediction. Several pitfalls with the use of these datasets have been identified in the literature, as well as useful guidelines for generating more effective decoy datasets. We contribute to this ongoing discussion an empirical assessment of several decoy datasets commonly used in experimental studies. RESULTS: We find that artefacts and sampling issues in the large majority of these data make it trivial to discriminate the native structure. This underlines that evaluation based on the rank/z-score of the native is a weak test of scoring function performance. Moreover, sampling biases present in the way decoy sets are generated or used can strongly affect other types of evaluation measures such as the correlation between score and root mean squared deviation (RMSD) to the native. We demonstrate how, depending on type of bias and evaluation context, sampling biases may lead to both over- or under estimation of the quality of scoring terms, functions or methods. AVAILABILITY: Links to the software and data used in this study are available at http://dbkgroup.org/handl/decoy_sets. PMID- 19297351 TI - The PickPocket method for predicting binding specificities for receptors based on receptor pocket similarities: application to MHC-peptide binding. AB - MOTIVATION: Receptor-ligand interactions play an important role in controlling many biological systems. One prominent example is the binding of peptides to the major histocompatibility complex (MHC) molecules controlling the onset of cellular immune responses. Thousands of MHC allelic versions exist, making determination of the binding specificity for each variant experimentally infeasible. Here, we present a method that can extrapolate from variants with known binding specificity to those where no experimental data are available. RESULTS: For each position in the peptide ligand, we extracted the polymorphic pocket residues in MHC molecules that are in close proximity to the peptide residue. For MHC molecules with known specificities, we established a library of pocket-residues and corresponding binding specificities. The binding specificity for a novel MHC molecule is calculated as the average of the specificities of MHC molecules in this library weighted by the similarity of their pocket-residues to the query. This PickPocket method is demonstrated to accurately predict MHC peptide binding for a broad range of MHC alleles, including human and non-human species. In contrast to neural network-based pan-specific methods, PickPocket was shown to be robust both when data is scarce and when the similarity to MHC molecules with characterized binding specificity is low. A consensus method combining the PickPocket and NetMHCpan methods was shown to achieve superior predictive performance. This study demonstrates how integration of diverse algorithmic approaches can lead to improved prediction. The method may also be used for making ligand-binding predictions for other types of receptors where many variants exist. PMID- 19297352 TI - Identifying functional modules using expression profiles and confidence-scored protein interactions. AB - MOTIVATION: Microarray-based gene expression studies have great potential but are frequently difficult to interpret due to their overwhelming dimensions. Recent studies have shown that the analysis of expression data can be improved by its integration with protein interaction networks, but the performance of these analyses has been hampered by the uneven quality of the interaction data. RESULTS: We present Co-Expression Zone ANalysis using NEtworks (CEZANNE), a novel confidence-based method for extraction of functionally coherent co-expressed gene sets. CEZANNE uses probabilities for individual interactions, which can be computed by any available method. We propose a probabilistic model and a weighting scheme in which the likelihood of the connectivity of a subnetwork is related to the weight of its minimum cut. Applying CEZANNE to an expression dataset of DNA damage response in Saccharomyces cerevisiae, we recover both known and novel modules and predict novel protein functions. We show that CEZANNE outperforms previous methods for analysis of expression and interaction data. AVAILABILITY: CEZANNE is available as part of the MATISSE software at http://acgt.cs.tau.ac.il/matisse. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online. PMID- 19297353 TI - Nephrocalcinosis: new insights into mechanisms and consequences. PMID- 19297354 TI - Active vitamin D and its analogue, 22-oxacalcitriol, ameliorate puromycin aminonucleoside-induced nephrosis in rats. AB - BACKGROUND: Recent studies have demonstrated that podocyte injury, which results in proteinuria, leads to tubulointerstitial fibrosis. Although some studies have revealed that vitamin D administration protects renal structure and function in mesangial cell proliferative and/or excessive matrix productive models, the effects of vitamin D on podocyte injury have remained uncertain. METHODS: In this study, we examined whether administration of active vitamin D (calcitriol) or its analogue, 22-oxacalcitriol (maxacalcitol), is preventative in podocyte injury using the puromycin aminonucleoside nephrosis model with neither mesangial proliferation nor matrix accumulation. RESULTS: Before the onset of proteinuria, renal 1alpha-hydroxylase and 24-hydroxylase were markedly down-regulated and up regulated, respectively, leading to impaired vitamin D activation. Thereafter, serum 25-hydroxyvitamin D decreased along with the increased excretion of vitamin D-binding protein in urine. After confirming that podocytes express vitamin D receptor and all retinoid X receptors (RXRs) except RXR-alpha, we found that daily administration of calcitriol or its analogue 22-oxacalcitriol ameliorated the nephrotic state by protecting podocytes, as shown by the reduced staining of desmin (podocyte injury marker) and the upregulation of nephrin and podocin. These data suggest that the impairment of the vitamin D system plays a role in increasing proteinuria in podocyte injury. CONCLUSIONS: We demonstrated the breakdown of the vitamin D activation system in podocyte injury, and established a preventative role for vitamin D in podocyte injury. PMID- 19297355 TI - Nephron number and individual glomerular volumes in male Caucasian and African American subjects. AB - BACKGROUND: Glomerular hypertrophy has been described in several populations at high risk of chronic kidney disease. Total nephron (and thereby glomerular) number (N(glom)) varies widely in normal adult human kidneys and is generally inversely correlated with mean glomerular volume (V(glom)). However, little is known about the range of individual glomerular volumes (IV(glom)) within single human kidneys and the association with N(glom). The aim of the present study was to estimate IV(glom) in Caucasian and African Americans and identify any associations between heterogeneity in IV(glom) and nephron number. METHODS: Using unbiased stereological techniques, IV(glom) was determined for 30 glomeruli in each of 24 adult male kidneys from Jackson, MS, USA (12 Caucasian and 12 African American). Half of each group had 'high' N(glom) (>1.2 million nephrons per kidney) and the other half had 'low' N(glom) (<600 000). RESULTS: Caucasians with high N(glom) had a relatively homogeneous distribution of IV(glom) as well as a relatively low mean value, while those with low N(glom) had much greater heterogeneity of IV(glom), as well as a larger IV(glom) (P < 0.0001) compared with those with high N(glom). This disparity was not apparent in African Americans, however, where subjects with both high and low N(glom) showed substantial heterogeneity in IV(glom) and larger mean values (P = 0.95). CONCLUSIONS: High N(glom) appeared to protect against glomerular enlargement and volume heterogeneity in Caucasians. However, substantial variation in IV(glom) and net enlargement in glomerular size in African Americans with high nephron numbers suggest that additional forces, independent of low N(glom), are driving glomerular enlargement and heterogeneity. PMID- 19297356 TI - Plasma phospholipid transfer protein, cholesteryl ester transfer protein and lecithin:cholesterol acyltransferase in end-stage renal disease (ESRD). AB - BACKGROUND: Chronic kidney disease (CKD) results in accelerated atherosclerosis that is primarily caused by inflammation, oxidative stress and impaired triglyceride and HDL metabolisms. Several plasma proteins including phospholipid transfer protein (PTLP), cholesteryl ester transfer protein (CETP) and lecithin:cholesterol acyltransferase (LCAT) affect HDL metabolism. PLTP transfers phospholipids and free cholesterol from triglyceride-rich lipoproteins to HDL, phospholipids between HDL particles and facilitates cholesterol efflux from cells. CETP catalyzes the transfer of cholesteryl esters from HDL to LDL in exchange for triglycerides, and LCAT catalyzes esterification of free cholesterol on the surface of HDL. Given the role of these proteins in the regulation of HDL metabolism, we examined the effect of ESRD on plasma PLTL, CETP and LCAT. METHODS: A group of 21 stable ESRD patients maintained on haemodialysis and a group of 21 age-matched normal control individuals were included in the study. Plasma apolipoprotein A-1, PLTP, CETP and LCAT levels were measured. RESULTS: Plasma triglyceride concentration was elevated and plasma HDL cholesterol, apolipoprotein A-1 and LCAT concentrations were significantly reduced, whereas plasma PLTP and CETP concentrations and activities were unchanged in the ESRD patients. CONCLUSIONS: These findings point to acquired LCAT and Apo A-1 deficiencies and tend to exclude dysregulation of PLTP or CETP in the pathogenesis of HDL abnormalities in haemodialysis patients. PMID- 19297357 TI - Chronic kidney disease, creatinine and cognitive functioning. AB - BACKGROUND: Non-dialysis-dependent chronic kidney disease (CKD) is related to cognitive impairment. Previous studies have not explored the extent of impairment across multiple cognitive domains. We examined the range of specific cognitive abilities affected by CKD and whether the associations of CKD with cognition were eliminated by statistical control for cardiovascular disease correlates of CKD. METHODS: We performed a community-based cross-sectional study with 923 individuals free from dementia and end-stage renal disease. Two groups were defined based on estimated glomerular filtration rate (eGFR): eGFR<60 mL/min/1.73 m(2) versus eGFR >or= 60 mL/min/1.73 m(2). Outcome measures were scores from multiple clinical tests of specific cognitive abilities. The GFR classifications and serum creatinine levels were related to measures of cognitive performance using logistic and linear regression analyses with three sets of covariates: (1) basic (age, education, gender and race); (2) basic+risk factors for cardiovascular disease (CVD) and (3) basic+risk factors for CVD+stroke. RESULTS: An eGFR <60 mL/min/1.73 m(2) was present in 142 (15.4%) individuals; the mean (SD) eGFR in this subgroup was 49.7 (10.7). CKD was related to lower cognitive performance despite adjustment for CVD risk factors (CVD-RF). Adjusting for CVD RF and stroke, odds ratios and 95% confidence intervals associated with performing in the lowest quartile of the distribution of the Global, Visual Spatial Organization/Memory and Scanning and Tracking scores for the eGFR < 60 group were 1.97 (1.25, 3.10); 1.88 (1.21, 2.93) and 1.83 (1.56, 2.87), P < 0.01 with eGFR >or= 60 group as the reference group. CONCLUSIONS: Global performance and specific cognitive functions are negatively affected early in CKD. Targeted screening for cognitive deficits in kidney disease patients early in their disease course may be warranted. PMID- 19297358 TI - Nutritional status evaluation and survival in haemodialysis patients in one centre from Romania. AB - BACKGROUND: Protein-energy wasting is a common complication and an important predictive factor for mortality in chronic dialysis patients. Therefore, nutritional status needs to be regularly assessed in these patients, by using several methods, and, if malnutrition is present, its possible causes should be thoroughly searched for and properly treated. MATERIAL AND METHODS: In 149 prevalent haemodialysis patients (82 men, mean age 53.9 +/- 13.7 years), we evaluated the nutritional status by anthropometrics [post-dialysis height (H), body weight (BW), body mass index (BMI), mid-arm circumference (MAC), tricipital skin-fold thickness (TST), mid-arm muscle circumference (MAMC), corrected mid-arm muscle area (cMAMA) and three-category subjective global assessment score (SGA)], biochemical tests [protein equivalent of nitrogen appearance (nPNA), and pre dialysis serum albumin, creatinine, total cholesterol, bicarbonate and haemoglobin (Hb) levels] and bioelectrical impedance analysis (BIA) to estimate body composition [percent body fat (%BF), fat-free mass (%FFM), body cell mass (%BCM), extracellular mass (%ECM) and the phase angle (PhA)]. RESULTS: Age was found to be positively correlated with BMI (P = 0.001), and inversely correlated with %BCM (P = 0.013). Patients with A-category SGA were significantly younger (50.1 versus 63.7 years) than those with B-category SGA. Patients with diabetes had lower %BCM (32.9 versus 35.9%; P = 0.035) and PhA (5.5 versus 6.9 degrees ; P = 0.0007) than those without diabetes. The presence of heart failure was associated with significantly reduced nPNA (1.17 versus 1.34 g/kg day; P = 0.014), MAMC (22.0 versus 23.6 cm(2); P = 0.041), %BCM (33.0 versus 36.1; P = 0.021), PhA (5.8 versus 7.0 degrees ; P = 0.031), serum albumin (39.7 versus 42.4 g/l; P = 0.013) and serum creatinine (8.1 versus 9.4 mg/dl; P = 0.010), and with a higher percent of B-category SGA (47.8% versus 22.6%; P = 0.019). Eleven deaths (7.4%) occurred during the follow-up period. Among general factors, age >or= 55, the presence of diabetes, and dialysis vintage <2 years were associated with significantly reduced survival. Among nutritional factors, B-category SGA, nPNA <1.2 g/kg day, %BF <15% and PhA <6 degrees significantly predicted mortality in both Kaplan-Meier and Cox analyses. The most important risk factor appeared to be nPNA; for every 0.1 g/kg day increase in nPNA, death risk decreased by 15%. CONCLUSIONS: In our haemodialysis patients, advancing age, diabetes and heart failure were associated with worse nutritional status, as estimated by anthropometry, biochemical markers and BIA. Age >or=55 years, the presence of diabetes, nPNA <1.2 g/kg day, lower SGA score, %BF <15% and PhA <6 degrees were associated with significantly increased death risk. PMID- 19297359 TI - The correlation between dental calculus and disturbed mineral metabolism in paediatric patients with chronic kidney disease. AB - BACKGROUND: Vascular calcifications have been documented in children with end stage renal disease. However, only a few reports have described abundant dental calculus formation in children suffering from chronic kidney disease (CKD). Moreover, dental calculus scores (DCS) and their correlation with renal disease severity have not been studied. METHODS: DCS in 74 young CKD patients were evaluated: 25 pre-dialytic (PrD), 18 on dialysis (D) and 31 with transplants (T) compared to 32 healthy participants (C). Saliva and serum analysis included creatinine (Cr), urea (U), calcium (Ca), phosphorous (P), magnesium (Mg) as well as intraoral pH levels. RESULTS: All patient groups presented high DCS. DCS and pH levels were higher in the D group with a positive correlation between pH and lower incisor DCS (r = 0.56, P = 0.017). The highest salivary Ca was found in the PrD group. Salivary P in the PrD group was found to be higher than in the T and C groups. The lowest salivary Mg was found in the D group while the highest salivary Ca x P product was found in the PrD group. In all patient groups, salivary U was higher than in the C group with a 2.5-fold increase in the D group. Salivary Cr resembled the U salivary concentrations. CONCLUSIONS: Alterations in salivary Ca, P, Mg, U, Cr and intraoral pH levels were observed in the patient groups. DCS correlated with renal disease severity and therefore may be a reflection of other tissue calcification pathologies found in these patients. PMID- 19297360 TI - Treatment of type 2 diabetes in chronic kidney disease: meekly follow the herd or call to arms? PMID- 19297361 TI - This extraordinary extramedullary haematopoiesis. PMID- 19297362 TI - Antifibrotic effects of pioglitazone on the kidney in a rat model of type 2 diabetes mellitus. AB - BACKGROUND: Recent evidence suggests that treatment of type 2 diabetes with thiazolidinediones [peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonists], ameliorates glomerulosclerosis and tubulointerstitial fibrosis in the rat kidney. In the current work, we have investigated whether these drugs, and specifically pioglitazone (PGT), act by preventing fibrosis and kidney dysfunction mainly through antioxidant and anti-inflammatory effects, independently of glycaemic control. METHODS: Male 2- to 3-month-old obese Zucker fa/fa (OZR) and ZDF fa/fa rats (ZDFR), and their control the lean Zucker rat (LZR), were used. Diabetic rats were given either a low dose (0.6 mg/kg/day) or a high dose (12 mg/ kg/day) of PGT in the chow for 2 or 4-5 months. Glycaemia, blood pressure, creatinine clearance and proteinuria were determined, and the underlying histopathology was defined with markers of fibrosis, glomerular damage, oxidative stress and inflammation by immunohistochemistry/ quantitative image analysis in tissue sections, and western blots and ad hoc assays in fresh tissue. RESULTS: PGT at low doses given for 4-5 months considerably reduced blood pressure, proteinuria and creatinine clearance. This was associated with amelioration of renal tissue damage and fibrosis, evidenced by the glomerulosclerosis, tubulointerstitial fibrosis, tubular atrophy and podocyte injury indexes, and of oxidative stress and inflammation, as shown by the decrease in the respective markers, although glycaemia remained high and obesity was not affected. CONCLUSIONS: These results indicate that low doses of PGT ameliorate renal fibrosis and preserve renal function in this animal model of metabolic syndrome, independently of glycaemic control or effects on body weight. PMID- 19297363 TI - Predictive value of obstructive sleep apnoea assessed by the Berlin Questionnaire for outcomes after the catheter ablation of atrial fibrillation. AB - AIMS: Obstructive sleep apnoea (OSA) is a risk factor for atrial fibrillation (AF) recurrence after catheter ablation. The purpose of this study is to determine whether OSA assessed by the simple tool, the Berlin Questionnaire (BQ), is useful in predicting the efficacy of catheter ablation of AF. METHODS AND RESULTS: The patient population consisted of 210 consecutive patients [mean age 58 +/- 10 years, 167 (80%) male, 119 (57%) paroxysmal] who underwent catheter ablation of AF and completed the BQ. Clinical success was defined as at least 90% reduction in AF burden after 3-month blanking period. Mean duration of follow-up was 25 +/- 12 months. One hundred and one of the 118 (85%) patients at low risk for OSA had clinical success as opposed to 64 of the 92 (70%) patients at high risk for OSA on BQ (P = 0.005). On multivariate analysis, only high-risk for OSA on BQ emerged as an independent predictor of procedural failure (OR 4.53, CI: 1.21-16.87, P = 0.02). CONCLUSION: High risk of OSA on BQ predicts procedural failure after catheter ablation of AF. We recommend the use of BQ for risk stratifying patients for OSA prior to AF ablation procedures and to identify patients for formal sleep study assessment. PMID- 19297364 TI - Precise electrocardiographic measurements and clinical sense. PMID- 19297365 TI - In the field of cardiac resynchronization therapy is left ventricular pacing via the coronary sinus a mature technique. PMID- 19297366 TI - A dynamic model of plant growth with interactions between development and functional mechanisms to study plant structural plasticity related to trophic competition. AB - BACKGROUND AND AIMS: The strong influence of environment and functioning on plant organogenesis has been well documented by botanists but is poorly reproduced in most functional-structural models. In this context, a model of interactions is proposed between plant organogenesis and plant functional mechanisms. METHODS: The GreenLab model derived from AMAP models was used. Organogenetic rules give the plant architecture, which defines an interconnected network of organs. The plant is considered as a collection of interacting 'sinks' that compete for the allocation of photosynthates coming from 'sources'. A single variable characteristic of the balance between sources and sinks during plant growth controls different events in plant development, such as the number of branches or the fruit load. KEY RESULTS: Variations in the environmental parameters related to light and density induce changes in plant morphogenesis. Architecture appears as the dynamic result of this balance, and plant plasticity expresses itself very simply at different levels: appearance of branches and reiteration, number of organs, fructification and adaptation of ecophysiological characteristics. CONCLUSIONS: The modelling framework serves as a tool for theoretical botany to explore the emergence of specific morphological and architectural patterns and can help to understand plant phenotypic plasticity and its strategy in response to environmental changes. PMID- 19297367 TI - The heart requires glycerol as an energy substrate through aquaporin 7, a glycerol facilitator. AB - AIMS: Cardiomyocytes require fatty acids and glucose for energy production. However, other nutrients and substrates that may serve as possible candidates for a cardiac energy source have not been fully studied. Several reports showed that a moderate expression of aquaporin 7 (AQP7), a member of the aquaglyceroporin family that is permeated by glycerol and water, is observed in heart tissue. However, the functional role of cardiac AQP7 is not clear. The aim of this study was to investigate the significance of glycerol as a cardiac energy substrate and to clarify the role of cardiac AQP7. METHODS AND RESULTS: Heart function and morphology were examined in AQP7-knockout (KO) mice under basal conditions and during pressure overload [isoproterenol infusion and transverse aortic constriction (TAC)]. Glycerol uptake and glycerol-dependent ATP production were measured in AQP7-knockdown cardiac cells. Cardiac glycerol consumption was analysed in ex vivo beating hearts. Cardiac morphology and function in KO mice were similar to those of wild-type (WT) mice under basal conditions, although low glycerol and ATP content were noted in hearts of KO mice. In H9c2 cardiomyotubes, knockdown of AQP7 was associated with a significant reduction of glycerol uptake. The ex vivo heart study demonstrated that cardiac glycerol consumption levels in KO mice were significantly lower than those of WT mice. Furthermore, isoproterenol challenge induced severe left ventricular hypertrophy in KO mice, and TAC resulted in a higher mortality rate in KO mice than in WT mice. CONCLUSION: The results indicate that AQP7 acts as a glycerol facilitator in cardiomyocytes and that glycerol is a substrate for cardiac energy production. PMID- 19297369 TI - Lipid signalling in cardiovascular pathophysiology. PMID- 19297368 TI - Angiopoietin-1 alters microvascular permeability coefficients in vivo via modification of endothelial glycocalyx. AB - AIMS: In this study, we wished to determine whether angiopoietin-1 (Ang1) modified the permeability coefficients of non-inflamed, intact continuous, and fenestrated microvessels in vivo and to elucidate the underlying cellular mechanisms. METHODS AND RESULTS: Permeability coefficients were measured using the Landis-Michel technique (in frog and rat mesenteric microvessels) and an oncopressive permeability technique (in glomeruli). Ang1 decreased water permeability (L(P): hydraulic conductivity) in continuous and fenestrated microvessels and increased the retention of albumin (sigma: reflection coefficient) in continuous microvessels. Endothelial glycocalyx is common to these anatomically distinct microvascular beds, and contributes to the magnitude of both L(P) and sigma. Ang1 treatment increased the depth of endothelial glycocalyx in intact microvessels and increased the content of glycosaminoglycan of cultured microvascular endothelial cell supernatant. Ang1 also prevented the pronase-induced increase in L(P) (attributable to selective removal of endothelial glycocalyx by pronase) by restoration of glycocalyx at the endothelial cell surface. The reduction in permeability was inhibited by a cell transport inhibitor, Brefeldin. CONCLUSION: Ang1 modifies basal microvessel permeability coefficients, in keeping with previous reports demonstrating reduced solute flux in inflamed vessels. Anatomical, biochemical, and physiological evidence indicates that modification of endothelial glycocalyx is a novel mechanism of action of Ang1 that contributes to these effects. PMID- 19297370 TI - Helium-induced late preconditioning in the rat heart in vivo. AB - BACKGROUND: A recent study showed that the noble gas helium induces early myocardial preconditioning. Cyclooxygenase-2 (COX-2) has been shown to be an important mediator in the signal transduction of late preconditioning. In the present study, we investigated whether helium induces late preconditioning in a concentration-dependent, time-dependent, or in both manner and whether COX-2 activity, mitochondrial function, or both are involved. METHODS: The study was performed in male Wistar rats and consisted of two parts. In part 1, late preconditioning was achieved by administration of 70%, 50%, 30%, and 10% helium for 15 min 24 h before ischaemia/reperfusion (I/R). Based on the findings of part 1, in additional experiments 30% helium was administered subsequently three and two days before I/R. Furthermore, additional rats were pretreated with the COX-2 inhibitor NS-398 (5 mg kg(-1)) with and without 30% helium. Additional experiments were performed for mitochondrial analysis. RESULTS: Helium concentrations of 70%, 50%, and 30% but not 10% reduced infarct size [He-LPC 70: 37(13)%, He-LPC 50: 34(16)%, He-LPC 30: 40(9)%; each P<0.05 vs CONTROL: 55(8)%, He-LPC 10: 53(4)%; P>0.05 vs CONTROL]. Repeated administration of helium did not further enhance cardioprotection. NS-398 completely abolished cardioprotection by 30% helium [He-LPC 30+NS-398: 57(9)%; P<0.05 vs He-LPC 30] but had itself no effect on infarct size [NS-398: 55(9)%; P>0.05 vs CONTROL]. There were no differences in mitochondrial function after helium preconditioning. CONCLUSIONS: Helium induces late preconditioning. Cardioprotection is already maximal with administration of one cycle of 30% helium and is abolished by functional blockade of COX-2 activity. PMID- 19297371 TI - Evaluation of the predictive performance of four pharmacokinetic models for propofol. AB - BACKGROUND: This study has compared the predictive performance of four pharmacokinetic models, two of which are currently incorporated in commercial target-controlled infusion pumps for the administration of propofol. METHODS: Arterial propofol concentrations and patient characteristic data were available from nine patients who, in a published study, had received a standardized infusion of propofol. Predicted concentrations with 'Diprifusor' (Marsh), 'Schnider', 'Schuttler', and 'White' models were obtained by computer simulation. The predictive performance of each model was assessed overall and over the following phases: rapid infusion (1-5 min), early (1-21 min), maintenance (21-min end-infusion), and recovery (2-20 min post-infusion). RESULTS: The overall assessment, based on 29-36 samples from each patient, indicated that all four models were clinically acceptable. However, the negligible bias (-0.1%) with the 'Schnider' model was accompanied by overprediction in the rapid infusion phase and underprediction during recovery. This changing bias over time was not detected as 'divergence' when assessed on absolute performance error (APE), (1.4% h(-1)) but became significant (13.2% h(-1)) when based on changes in signed PE over time. The 'Schuttler' model performed well at most phases but overpredicted concentrations during recovery. The White model led to a marginal improvement over 'Diprifusor' and would be expected to reduce the positive bias usually seen with 'Diprifusor' systems. CONCLUSIONS: In assessing the predictive performance of pharmacokinetic models, additional information can be obtained by analysis of bias at different phases of an infusion. The evaluation of divergence should involve linear regression analysis of both absolute and signed PEs. PMID- 19297372 TI - Predicting the onset of delirium in the post-operative patient. AB - The number of people over 65 is increasing and those over 65 requiring surgery will likewise increase. Post-operative delirium (POD) affects up to 47% of patients undergoing surgery and is more prevalent in older people. Importantly, POD is associated with increased morbidity, mortality, length of stay and care home placement. There is evidence that specialist geriatrician input reduces POD but to be cost effective, needs to target patients with increased risk for POD. Many factors have been associated with increased risk of POD, including age, cognitive impairment, gender, depression, alcohol, drug use, smoking, co morbidity, functional status, ASA score and pre-operative biochemical and haematological abnormalities. This article reviews the literature associated with the above factors, considers frailty as a factor and also suggests that POD may be associated with rapidity of onset and severity of the insult to the patient. PMID- 19297373 TI - Higher level gait disorders in subcortical chronic vascular encephalopathy: a single photon emission computed tomography study. AB - BACKGROUND: the so-called higher level gait disorders include several types of gait disorders in which there are no major modifications in strength, tone, sensitivity, coordination and balance. Brain activation sites related to walking have been investigated using SPECT in humans. The aim of the study was to investigate brain activation during walking in subjects with high-level gait disorders due to chronic subcortical vascular encephalopathy. SUBJECTS: twelve patients with a chronic vascular encephalopathy were enrolled in the study. Seven subjects had apraxic gait while in the other five the gait was normal. All patients had undergone a recent cerebral magnetic resonance that revealed diffused chronic ischemic lesions within the white matter. METHODS: all 12 patients underwent a regional cerebral blood flow (rCBF) brain SPECT study with (99m)Tc-Bicisate on two separate days and under two different conditions: at rest (baseline) and while walking (functional). RESULTS: the rCBF increase induced by the treadmill test (functional-baseline), bilaterally in the medial frontal gyrus and in the anterior lobes of the cerebellum, resulted significantly (P < 0.001) lower in patients with gait apraxia versus those without it. CONCLUSIONS: this study of the brain with SPECT records the areas of perfusion deficit that appear in apraxic subjects when they walk, compared with the recordings obtained with the same investigation performed at rest. PMID- 19297374 TI - Vitamin D and in-patient falls. PMID- 19297375 TI - Akinetic mutism--not coma. AB - We describe a case of akinetic mutism mistaken as coma. A 77-year old lady presented with apparent unresponsiveness. In fact she responded when stimulated. She subsequently developed movements typical of clonic perseveration, erroneously treated as seizures. She also had features of a frontal lobe syndrome. Initial CT scan showed no abnormality; it was only after an MRI scan that a diagnosis of bilateral paramedian thalamic infarction was made. MRI scanning should be considered early in the investigation of patients with atypical neurological presentation. PMID- 19297376 TI - Comparative analysis of extended-spectrum-{beta}-lactamase-carrying plasmids from different members of Enterobacteriaceae isolated from poultry, pigs and humans: evidence for a shared {beta}-lactam resistance gene pool? PMID- 19297377 TI - Effect of long-term trimethoprim/sulfamethoxazole treatment on resistance and integron prevalence in the intestinal flora: a randomized, double-blind, placebo controlled trial in children. AB - OBJECTIVES: The aim of this study was to test the hypothesis that trimethoprim/sulfamethoxazole selects for integron-positive and multidrug resistant Enterobacteriaceae in the intestinal flora. METHODS: During 1 year of follow-up, antibiotic susceptibility and the presence of integrons were determined in faecal Enterobacteriaceae isolated from 99 children with chronic active otitis media, randomly assigned to treatment with trimethoprim/sulfamethoxazole or placebo (http://www.clinicaltrials.gov/; trial registration number NCT00189098). RESULTS: At 6 and 12 weeks of follow-up, 32 (91%) and 24 (67%) children in the trimethoprim/sulfamethoxazole group carried trimethoprim/sulfamethoxazole-resistant Enterobacteriaceae versus 10 (21%) and 8 (17%) children in the placebo group [rate differences (RDs): 70 (95% CI: 55; 85) and 50 (95% CI: 31; 69)], respectively. Multiresistance also increased during trimethoprim/sulfamethoxazole treatment. At 6 weeks of follow-up, the integron prevalence was 26 (79%) in the trimethoprim/sulfamethoxazole group and 10 (22%) in the placebo group [RD: 57 (95% CI: 39; 75)]. After 12 weeks the integron prevalence, and after 1 year the susceptibility levels, had returned to baseline values. CONCLUSIONS: Initially, trimethoprim/sulfamethoxazole usage was strongly associated with the appearance of integron-positive (multi)drug-resistant Enterobacteriaceae in the intestinal flora. After prolonged exposure to trimethoprim/sulfamethoxazole, however, this population of Enterobacteriaceae was substituted by a population with non-integron-associated resistance mechanisms. After trimethoprim/sulfamethoxazole was discontinued, susceptibility rates to all antibiotics returned to baseline levels. PMID- 19297378 TI - Characterization of fluoroquinolone-resistant Shigella flexneri in Hangzhou area of China. AB - OBJECTIVES: The aim of this study was to characterize fluoroquinolone-resistant Shigella and determine whether the qnr and aac(6')-Ib-cr genes could contribute to sporadic shigellosis at the clinic in the Hangzhou area of China. METHODS: A total of 202 strains of Shigella (79 Shigella sonnei and 123 Shigella flexneri ) isolated from sporadic cases of shigellosis from 1998 to 2007 were analysed for their antimicrobial susceptibility. The gyrA, gyrB, parC, parE, qnr and aac(6') Ib-cr genes and the profiles and incompatibility of plasmids were characterized. Chromosomal DNA fingerprinting was determined by XbaI-based digestion and PFGE. RESULTS: All strains of S. sonnei were susceptible to fluoroquinolones (ciprofloxacin and levofloxacin) while 15 out of 123 strains of S. flexneri were resistant. All of the 15 resistant strains displayed common mutations in the gyrA and parC genes and formed eight distinct groups with unique molecular characteristics. Notably, 10 isolates showed mutations at codon 87 of gyrA, and the other 5 were qnrS-positive. Two strains were positive for the aac(6')-Ib-cr gene. Importantly, this is the first report of qnrS- and aac(6')-Ib-cr-positive Shigella in China, the qnrS-positive S. flexneri serotypes 1a, 2a and 4c and the aac(6')-Ib-cr-positive S. flexneri serotypes 2a and 4c worldwide. CONCLUSIONS: The common mutations at position 83 of gyrA and position 80 of parC were crucial for resistance to nalidixic acid in S. flexneri. The mutation at position 87 of gyrA or the presence of the qnrS gene is necessary for high-level resistance to fluoroquinolones in Shigella isolates from China. PMID- 19297379 TI - Novel CTX-M {beta}-lactamase genotype distribution and spread into multiple species of Enterobacteriaceae in Changsha, Southern China. AB - OBJECTIVES: The aim of this study was to undertake a survey of the occurrence of CTX-M and SHV extended-spectrum beta-lactamase (ESBL) genotypes in Enterobacteriaceae from Hunan Province, China. METHODS: Clinical isolates (425) from three major hospitals in Changsha, Hunan Province, were collected between October 2004 and July 2005, and their antimicrobial susceptibilities of the genotype of bla(CTX-M) and bla(SHV) were determined. Random amplified polymorphic DNA was used to characterize the clonality of all of the isolates. RESULTS: The overall rate of ESBL-positive isolates was 33.4% (142/425). The dominant ESBLs were CTX-M types, and were found in 109/142 (76.8%) isolates comprising seven different genera/species, namely Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Proteus vulgaris and Providencia stuartii. The most common bla(CTX-M) genotypes were bla(CTX-M-14) (47.7%), bla(CTX-M-3) (29.4%) and bla(CTX-M-15) (17.4%). A novel gene derived from bla(CTX-M-15), bla(CTX-M-82) (Ala-40-->Pro), was identified. CONCLUSIONS: The dominant ESBL genotype in Hunan Province was bla(CTX-M). The high prevalence (17.4%) of bla(CTX-M-15) has not previously been reported from China. Our results identify that an epidemic of bla(CTX-M) in Changsha, Hunan Province, has evolved with the appearance and spread of bla(CTX-M-15) against the dominant genotypes bla(CTX-M-14) and bla(CTX-M-3.) The worldwide dominance of bla(CTX-M-15) could be poised to spread to China, displacing the current prevailing genotypes. PMID- 19297380 TI - The role of AA sponsors: a pilot study. AB - AIMS: The aim of this study was to explore the roles of Alcoholics Anonymous (AA) sponsors and to describe the characteristics of a sample of sponsors. METHODS: Twenty-eight AA sponsors, recruited using a purposive sampling method, were administered an unstructured qualitative interview and standardized questionnaires. The measurements included: a content analysis of sponsors' responses; Severity of Alcohol Dependence Questionnaire-Community version (SADQ C) and Alcoholics Anonymous Affiliation Scale (AAAS). RESULTS: Sample characteristics were as follows: the median length of AA attendance was 9.5 years (range 5-28); the median length of sobriety was 11 years (range 4.5-28); the median number of sponsees per sponsor was 1 but there was a wide range (0-17, interquartile range 3.75); and the sponsors were highly affiliated to AA (median AAAS score 8.75, range 5.5-8.75, maximum possible score 9). Past alcohol dependence scores were surprisingly low: 5 (18%) sponsors had mild, 14 (50%) moderate and 9 (32%) severe dependence according to the SADQ-C (median 26.5, range 11-56). Sponsorship roles were as follows: 16 roles were identified through the initial content analysis. These were distilled into three super-ordinate roles through a thematic analysis: (1) encouraging sponsees to work the programme of AA (doing the 12 steps and engaging in AA activity); (2) support (regular contact, emotional support and practical support); and (3) carrying the message of AA (sharing sponsor's personal experience of recovery with sponsees). CONCLUSIONS: The roles identified broadly corresponded with the AA literature delineating the duties of a sponsor. This non-random sample of sponsors was highly engaged in AA activity but only had a past history of moderate alcohol dependence. PMID- 19297382 TI - Rocking will tell it. PMID- 19297381 TI - Drug discovery in psychiatric illness: mining for gold. AB - The discovery of truly efficacious treatments that lead to full recovery is a daunting task in psychiatric illness. A systems-based orientation to in vivo pharmacology has been suggested as a way to transform psychiatric drug discovery and development. A critical catalyst in the success of recent systems biology efforts has been the incorporation of data mining strategies. Our approach to the drug discovery problem has been to utilize the whole animal to provide a systems response that is subsequently mined for predictive attributes with known psychopharmacological value. Our in vivo data mining approach, termed Pattern Array, establishes a framework for screening novel chemical entities based upon a response that represents the net pharmacological effect on the system of interest, namely the central nervous system (CNS). Large scale screening of small molecules by non-conventional approaches such as this at a systems level may improve the identification of novel chemical entities with psychiatric utility. This type of approach will compliment the more labor-intensive models based upon construct validity. It will take the collective effort of many disciplines and numerous strategies in close association with clinical colleagues to address quality of life issues and breakthrough treatment barriers in psychiatric illness. PMID- 19297383 TI - A diagnostic odyssey: young woman with chest pain. PMID- 19297385 TI - Association of plasma asymmetrical dimethylarginine (ADMA) with elevated vascular superoxide production and endothelial nitric oxide synthase uncoupling: implications for endothelial function in human atherosclerosis. AB - BACKGROUND: Asymmetrical dimethylarginine (ADMA), an endogenous inhibitor of endothelial nitric oxide synthase (eNOS), is considered to be a risk factor for atherosclerosis. However, the mechanisms relating ADMA with vascular function have been evaluated in vitro and in animal models, but its effect in human vasculature is unclear. AIMS: We examined the impact of serum ADMA on endothelial nitric oxide (NO) bioavailability and vascular superoxide radical (O2-) production in patients with advanced atherosclerosis. METHODS AND RESULTS: Paired samples of saphenous veins (SVs) and internal mammary arteries (IMAs) were collected from 201 patients undergoing coronary bypass surgery, and serum ADMA was measured pre-operatively. The vasomotor responses of SV segments to acetylcholine (ACh) and bradykinin (Bk) were evaluated ex vivo. Vascular O2- was measured in paired SV and IMA by lucigenin-enhanced chemiluminescence. The l-NAME inhibitable as well as the NADPH-stimulated vascular O2- generation was also determined by chemiluminescence. High serum ADMA levels were associated with decreased vasorelaxation of SV to ACh (P < 0.05) and Bk (P < 0.05). Similarly, high serum ADMA was associated with higher total O2- production in both SVs and IMAs (P < 0.05) and greater L-NAME-inhibitable vascular O2- (P < 0.05). However, serum ADMA was not associated with NADPH-stimulated vascular O2-. In multivariable linear regression, serum ADMA was independently associated with vascular O2- in both SVs [beta (SE): 0.987 (0.412), P = 0.019] and IMAs [beta (SE): 1.905 (0.541), P = 0.001]. Asymmetrical dimethylarginine was also independently associated with maximum vasorelaxation in response to both ACh [beta (SE): 14.252 (3.976), P = 0.001] and Bk [beta (SE): 9.564 (3.762), P = 0.013]. CONCLUSION: This is the first study that demonstrates an association between ADMA and important measures of vascular function, such as vascular O2- production and NO bioavailability directly in human vessels. Although serum ADMA has no effect on NADPH-stimulated superoxide in intact vessels, it is associated with greater eNOS uncoupling in the human vascular endothelium of patients with coronary artery disease. PMID- 19297384 TI - Socioeconomic status, blood pressure progression, and incident hypertension in a prospective cohort of female health professionals. AB - AIMS: The aim of this study was to examine the association between socioeconomic status, blood pressure (BP) progression, and incident hypertension. METHODS AND RESULTS: We included 27 207 female health professionals free of hypertension and cardiovascular disease at baseline. Participants were classified into five education and six income categories. The main outcome variables were BP progression at 48 months of follow-up and incident hypertension during the entire study period. At 48 months, 48.1% of women had BP progression. The multivariable adjusted relative risks [95% confidence intervals (CIs)] for BP progression were 1.0 (referent), 0.96 (0.92-1.00), 0.92 (0.88-0.96), 0.90 (0.85-0.94), and 0.84 (0.78-0.91) (P for trend <0.0001) across increasing education categories and 1.0 (referent), 1.01 (0.94-1.08), 0.99 (0.93-1.06), 0.97 (0.91-1.04), 0.96 (0.90 1.03), and 0.89 (0.83-0.96) across increasing income categories (P for trend = 0.0001). During a median follow-up of 9.8 years, 8248 cases of incident hypertension occurred. Multivariable adjusted hazard ratios (95% CI) were 1.0 (referent), 0.92 (0.86-0.99), 0.85 (0.79-0.92), 0.87 (0.80-0.94), and 0.74 (0.65 0.84) (P for trend <0.0001) across increasing education categories and 1.0 (referent), 1.07 (0.95-1.21), 1.07 (0.95-1.20), 1.06 (0.94-1.18), 1.04 (0.93 1.16), and 0.93 (0.82-1.06) (P for trend 0.08) across increasing income categories. In joint analyses, education but not income remained associated with BP progression and incident hypertension. CONCLUSION: Socioeconomic status, as determined by education but not by income, is a strong independent predictor of BP progression and incident hypertension in women. PMID- 19297386 TI - Apical transverse motion as surrogate parameter to determine regional left ventricular function inhomogeneities: a new, integrative approach to left ventricular asynchrony assessment. AB - AIMS: Left ventricular (LV) asynchrony assessment is mostly based on delays between regional myocardial velocity peaks. Regional function is barely considered. We propose apical transverse motion (ATM) as a new parameter integrating both temporal and functional information, which was tested in different conduction delays. METHODS AND RESULTS: We examined 67 patients, 11 patients with post-infarct ischaemic left bundle branch blocks (iLBBB) and 25 patients with non-ischaemic left bundle branch block (nLBBB), 12 patients with right bundle branch block (RBBB), and 19 normal healthy volunteers (NORM). Longitudinal colour tissue Doppler data were used to calculate the total transverse apex motion (ATM), the transverse motion in the four-chamber view plane alone (ATM(4CV)) as well as regional myocardial deformation and conventional LV asynchrony parameters. Median ATM was 1.8 mm in NORM, 1.5 mm in RBBB (P = 0.999), 2.4 mm in iLBBB (P = 0.183), and 4.3 mm in nLBBB (P < 0.001 vs. NORM and RSB). ATM(4CV) behaved similarly, showed a good correlation with regional deformation data, and distinguished well between NORM and LBBB (AUC = 0.87). CONCLUSION: Apical transverse motion is a new and simple parameter integrating information on both regional and temporal function inhomogeneities of the LV. It has a potential role in assessing LV asynchrony in the clinical context. PMID- 19297387 TI - Complex cardiovascular actions of alpha-adrenergic receptors expressed in the nucleus tractus solitarii of rats. AB - Although both alpha(1)- and alpha(2)-adrenergic receptors (ARs) are known to be expressed in the nucleus of the solitary tract (NTS), the functional significance of these receptors is still not fully established. In this study, we microinjected alpha(1)- and alpha(2)-AR agonists into the NTS of urethane anaesthetized Wister rats to study the cardiovascular effects in response to their activation. When the alpha(1)-AR agonist phenylephrine was microinjected into the area where barosensitive neurons are dominantly located (baro-NTS), mean arterial pressure (MAP) and heart rate (HR) were significantly elevated. When tested in the area where chemosensitive neurons are dominantly located (chemo NTS), however, MAP and HR were significantly decreased. Pretreatment with the non specific alpha-AR antagonist phentolamine into the NTS inhibited the phenylephrine-induced cardiovascular responses. In contrast, microinjection of the alpha(2)-AR agonist clonidine into either the baro-NTS or the chemo-NTS decreased MAP and HR; they were also inhibited by the alpha(2)-adrenergic antagonist yohimbine. Moreover, we immunohistochemically identified that cardiovascular responses induced by alpha(1)-ARs may be mediated by NTS neurons while those induced by alpha(2)-ARs may be mediated by astrocytes located in the barosensitive and chemosensitive areas of the NTS. These results suggest that both types of alpha-AR expressed in the NTS may be involved in regulating cardiovascular homeostasis via modulation of input signals from baroreceptor and chemoreceptor afferents; however, cardiovascular responses produced by stimulation of alpha(1)-ARs are strictly location specific within the NTS. PMID- 19297388 TI - Global deletion of thrombospondin-1 increases cardiac and skeletal muscle capillarity and exercise capacity in mice. AB - Thrombospondin-1 (TSP-1) is a known inhibitor of angiogenesis; however, a skeletal muscle phenotype of TSP-1 null mice has not been investigated. The purposes of this study were to compare and contrast TSP-1 null and wild-type mice by examining the following: (1) capillarity in the skeletal and cardiac muscles; (2) fibre type composition and oxidative enzyme activity in the hindlimb; and (3) the consequences of TSP-1 gene deletion for exercise capacity. In TSP-1 null mice, maximal running speed was 11% greater and time to exhaustion during submaximal endurance running was 67% greater compared with wild-type mice. Morphometric analyses revealed that TSP-1 null mice had higher (P < 0.05) capillarity in the heart and skeletal muscle than wild-type mice, whereas no differences for fibre type composition or oxidative enzyme activity were present between the two groups. Cardiac function, as measured by transthoracic echocardiography, revealed no difference in myocardial contractility but greater left ventricular end-diastolic and systolic dimensions, corresponding to an elevated heart mass in the TSP-1 null mice. The results of this study indicate that TSP-1 is an important endogenous negative regulator of angiogenesis that prevents excessive capillarization in the heart and skeletal muscles. The increased capillarity alone was sufficient to increase (P < 0.05) exercise capacity. These data demonstrate that the capillary-to-muscle interface is a critical factor that limits oxygen transport during exercise. PMID- 19297389 TI - The role of mammalian cardiac t-tubules in excitation-contraction coupling: experimental and computational approaches. AB - The sarcolemmal membrane of mammalian cardiac ventricular myocytes is characterized by the presence of invaginations called transverse tubules (t tubules). Transverse tubules occur at the Z-line as transverse elements with longitudinal extensions. While the existence of t-tubules has been known for some time, recent experimental studies have suggested that their structure and function are more complex than previously believed. There are, however, aspects of t-tubule function that are not currently amenable to experimental investigation, but can be investigated using computational and mathematical approaches. Such studies have helped elucidate further the possible role of t tubules in cell function. This review summarizes recent experimental and complementary computational studies which highlight the important role of t tubules in cardiac excitation-contraction coupling. PMID- 19297390 TI - PGC-1alpha/beta induced expression partially compensates for respiratory chain defects in cells from patients with mitochondrial disorders. AB - Members of the peroxisome proliferator-activated receptor gamma coactivator (PGC) family are potent inducers of mitochondrial biogenesis. We have tested the potential effect of increased mitochondrial biogenesis in cells derived from patients harboring oxidative phosphorylation defects due to either nuclear or mitochondrial DNA mutations. We found that the PGC-1alpha and/or PGC-1beta expression improved mitochondrial respiration in cells harboring a complex III or IV deficiency as well as in transmitochondrial cybrids harboring mitochondrial encephalomyopathy lactic acidosis and stroke A3243G tRNA((Leu)UUR) gene mutation. The respiratory function improvement was found to be associated with increased levels of mitochondrial components per cell, although this increase was not homogeneous. These results reinforce the concept that increased mitochondrial biogenesis is a promising venue for the treatment of mitochondrial diseases. PMID- 19297395 TI - Copy number variants, diseases and gene expression. AB - Copy number variation (CNV) has recently gained considerable interest as a source of genetic variation likely to play a role in phenotypic diversity and evolution. Much effort has been put into the identification and mapping of regions that vary in copy number among seemingly normal individuals in humans and a number of model organisms, using bioinformatics or hybridization-based methods. These have allowed uncovering associations between copy number changes and complex diseases in whole-genome association studies, as well as identify new genomic disorders. At the genome-wide scale, however, the functional impact of CNV remains poorly studied. Here we review the current catalogs of CNVs, their association with diseases and how they link genotype and phenotype. We describe initial evidence which revealed that genes in CNV regions are expressed at lower and more variable levels than genes mapping elsewhere, and also that CNV not only affects the expression of genes varying in copy number, but also have a global influence on the transcriptome. Further studies are warranted for complete cataloguing and fine mapping of CNVs, as well as to elucidate the different mechanisms by which they influence gene expression. PMID- 19297396 TI - The genetics of chronic inflammatory diseases. AB - Chronic inflammatory diseases have been at the forefront of the new genome-wide association study era. Conditions such as coeliac disease, type 1 diabetes, Crohn's disease and ulcerative colitis have all benefited with multiple loci identified and replicated for each condition. As cohort sample numbers increase and researchers collaborate and share cohorts, common susceptibility loci are beginning to emerge between several diseases. Crohn's disease and coeliac disease both demonstrate considerable overlap in their common genetic susceptibility with other related conditions. These shared loci offer an insight into the biology of the conditions but still present researchers with the problem of attempting to identify the true causal variants. PMID- 19297397 TI - The power of the zebrafish for disease analysis. AB - The zebrafish is increasingly being used for the functional annotation of human disease genes as well as to model the cellular and molecular mechanisms underlying pathological processes. Various attributes of the organism lend themselves to these studies, in particular the ease with which gene function can be manipulated together with the ability to image transgenically labelled cells in living animals. This review highlights a few recent studies relating to myopathies and neurodegenerative diseases in human that exemplify some of the advantageous attributes of the zebrafish. It also discusses some of its current limitations and how these are being addressed through the transfer of technologies from other systems. PMID- 19297398 TI - Small regulatory RNAs in neurodevelopmental disorders. AB - Increasingly complex networks of small RNAs act through RNA interference pathway to regulate gene expression. Recent evidence suggests that both development and proper function of central nervous system require intricate spatiotemporal expression of a wide repertoire of small regulatory RNAs. Misregulation of these small regulatory RNAs could contribute to the abnormalities in brain development that are associated with neurodevelopmental disorders. Here, we will review recent progress made toward understanding roles of small regulatory RNAs in neurodevelopmental disorders and discuss the potential involvement of newly discovered classes of small RNAs in these disorders. PMID- 19297400 TI - Post-translational modifications of expanded polyglutamine proteins: impact on neurotoxicity. AB - Polyglutamine diseases are a family of nine neurodegenerative disorders caused by expansion in different genes of a CAG triplet repeat stretch, which encodes an elongated polyglutamine tract. This polyglutamine tract is thought to confer a toxic gain of function to the bearing proteins, which leads to late onset and progressive loss of neurons in specific regions of the central nervous system. The mechanisms underlying specificity for neuronal vulnerability remain enigmatic. One explanation is that the polyglutamine tract is not the only determinant of neurodegeneration and that protein context and post-translational events may also be crucial for pathogenesis. Here, we review how post translational modifications of the polyglutamine proteins contribute to modulate neurotoxicity. PMID- 19297399 TI - Short non-coding RNA biology and neurodegenerative disorders: novel disease targets and therapeutics. AB - Genomic studies in model organisms and in humans have shown that complexity in biological systems arises not from the absolute number of genes, but from the differential use of combinations of genetic programmes and the myriad ways in which these are regulated spatially and temporally during development, senescence and in disease. Nowhere is this lesson in biological complexity likely to be more apparent than in the human nervous system. Increasingly, the role of genomic non protein coding small regulatory RNAs, in particular the microRNAs (miRNAs), in regulating cellular pathways controlling fundamental functions in the nervous system and in neurodegenerative disease is being appreciated. Not only might dysregulated expression of miRNAs serve as potential disease biomarkers but increasingly such short regulatory RNAs are being implicated directly in the pathogenesis of complex, sporadic neurodegenerative disease. Moreover, the targeting and exploitation of short RNA silencing pathways, commonly known as RNA interference, and the development of related tools, offers novel therapeutic approaches to target upstream disease components with the promise of providing future disease modifying therapies for neurodegenerative disorders. PMID- 19297401 TI - Parkinson's disease: from monogenic forms to genetic susceptibility factors. AB - Research in Parkinson's disease (PD) genetics has been extremely prolific over the past decade. More than 13 loci and 9 genes have been identified, but their implication in PD is not always certain. Point mutations, duplications and triplications in the alpha-synuclein (SNCA) gene cause a rare dominant form of PD in familial and sporadic cases. Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are a more frequent cause of autosomal dominant PD, particularly in certain ethnic groups. Loss-of-function mutations in Parkin, PINK1, DJ-1 and ATP13A2 cause autosomal recessive parkinsonism with early-onset. Identification of other Mendelian forms of PD will be a main challenge for the next decade. In addition, susceptibility variants that contribute to PD have been identified in several populations, such as polymorphisms in the SNCA, LRRK2 genes and heterozygous mutations in the beta-glucocerebrosidase (GBA) gene. Genome-wide associations and re-sequencing projects, together with gene-environment interaction studies, are expected to further define the causal role of genetic determinants in the pathogenesis of PD, and improve prevention and treatment. PMID- 19297402 TI - Lessons learnt from large-scale exon re-sequencing of the X chromosome. AB - A candidate gene approach to identifying novel causes of disease is concept limiting and in the new era of high throughput sequencing there is now no need to restrict the experiment to a few interesting genes. We have recently completed a large-scale exon re-sequencing project using Sanger sequencing technology to analyse approximately 1 Mb of coding sequence of the X chromosome in probands from >200 families with various forms of intellectual disability. We review the lessons learnt from this experience. Comparing large data sets will certainly reveal pathogenic mutations in genes that were not possible to identify previously. However, the task of distinguishing pathogenic mutations from rare sequence variants is not easy and is the most substantial challenge to the next decade. High-throughput technology has the attraction of being cheap, fast and comprehensive but for projects that require detailed coverage of a genomic region at an exhaustive level they may require a combination of large-scale with a small scale follow-up of difficult regions to sequence. The number of rare truncating variants present in coding regions of the X chromosome that are not pathogenic was 1%. The importance of the quality of the starting material both clinically and molecularly and the number of sequence variants both rare and common that any one individual has across their coding sequence is discussed. PMID- 19297403 TI - From germline towards somatic mutations in the pathophysiology of vascular anomalies. AB - The localized structural abnormalities that arise during vasculogenesis, angiogenesis and lymphangiogenesis, the developmental processes which give rise to the adult vasculature, are collectively termed vascular anomalies. The last 2 years have seen an explosion of studies that underscore paradominant inheritance, the combination of inherited changes with somatic second-hits to the same genes, as underlying rare familial forms. Moreover, local, somatic genetic defects that cause some of the common sporadic forms of these malformations have been unraveled. This highlights the importance of assessing for tissue-based genetic changes, especially acquired genetic changes, as possible pathophysiological causes, which have been largely overlooked except in the area of cancer research. Large-scale somatic screens will therefore be essential in uncovering the nature and prevalence of such changes, and their downstream effects. The identification of disease genes combined with exhaustive, precise clinical delineations of the entire spectra of associated phenotypes guides better management and genetic counseling. Such a synthesis of information on functional and phenotypic effects will enable us to make and use animal models to test less invasive, targeted, perhaps locally administered, biological therapies. PMID- 19297404 TI - Down syndrome--recent progress and future prospects. AB - Down syndrome (DS) is caused by trisomy of chromosome 21 (Hsa21) and is associated with a number of deleterious phenotypes, including learning disability, heart defects, early-onset Alzheimer's disease and childhood leukaemia. Individuals with DS are affected by these phenotypes to a variable extent; understanding the cause of this variation is a key challenge. Here, we review recent research progress in DS, both in patients and relevant animal models. In particular, we highlight exciting advances in therapy to improve cognitive function in people with DS and the significant developments in understanding the gene content of Hsa21. Moreover, we discuss future research directions in light of new technologies. In particular, the use of chromosome engineering to generate new trisomic mouse models and large-scale studies of genotype-phenotype relationships in patients are likely to significantly contribute to the future understanding of DS. PMID- 19297406 TI - Molecular genetics of human pigmentation diversity. AB - The genetic basis underlying normal variation in the pigmentary traits of skin, hair and eye colour has been the subject of intense research directed at understanding the diversity seen both between and within human populations. A combination of approaches have been used including comparative genomics of candidate genes and the identification of regions of the human genome under positive selection, together with genome-wide and specific allele association studies. Independent selection for different pigmentation gene sets has been found between Asian, European and African populations. Several genome-wide association studies for pigmentation have now been conducted and identified single nucleotide polymorphism (SNP) markers in known, TYR, TYRP1, OCA2, SLC45A2, SLC24A5, MC1R, ASIP, KITLG and previously unknown SLC24A4, IRF4, TPCN2, candidate genes. The contribution of SNP polymorphisms present in populations from South Asia have been tested and alleles found at TYR, SLC45A2 and SLC24A5 can largely account for differences between those of darkest and lightest skin reflectance using a simple additive model. Skin and hair colour associations in Europeans are found within a range of pigmentation gene alleles, whereas blue-brown eye colour can be explained by a single SNP proposed to regulate OCA2 expression. Functional testing of variant alleles has begun to connect phenotype correlations with biological differences. Variant MC1R alleles show direct correlations between the biochemical signalling properties of the encoded receptor and the red-hair fair skin pigmentation phenotype. Direct testing of a range of clonal melanocyte cultures derived from donor skin tissue characterized for three causal SNPs within SLC45A2, SLC24A5 and OCA2 has assessed their impact on melanin content and tyrosinase enzyme activity. From a culmination of genetic and functional studies, it is apparent that a number of genes impacting melanosome biogenesis or the melanin biosynthetic pathway are candidates to explain the diversity seen in human pigmentation. PMID- 19297405 TI - Constitutional aneuploidy and cancer predisposition. AB - Constitutional aneuploidies are rare syndromes associated with multiple developmental abnormalities and the alterations in the risk for specific cancers. Acquired somatic chromosomal aneuploidies are the most common genetic aberrations in sporadic cancers. Thus studies of these rare constitutional aneuploidy syndromes are important not only for patient counseling and clinical management, but also for deciphering the mechanisms by which chromosomal aneuploidy affect cancer initiation and progression. Here we review the major constitutional aneuploidy syndromes and suggest some general mechanisms for the associated cancer predisposition. PMID- 19297408 TI - Swimming exercise at weaning improves glycemic control and inhibits the onset of monosodium L-glutamate-obesity in mice. AB - Swimming exercises by weaning pups inhibited hypothalamic obesity onset and recovered sympathoadrenal axis activity, but this was not observed when exercise training was applied to young adult mice. However, the mechanisms producing this improved metabolism are still not fully understood. Low-intensity swimming training started at an early age and was undertaken to observe glycemic control in hypothalamic-obese mice produced by neonatal treatment with monosodium l glutamate (MSG). Whereas MSG and control mice swam for 15 min/day, 3 days a week, from the weaning stage up to 90 days old, sedentary MSG and normal mice did not exercise at all. After 14 h of fasting, animals were killed at 90 days of age. Perigonadal fat accumulation was measured to estimate obesity. Fasting blood glucose and insulin concentrations were also measured. Fresh isolated pancreatic islets were used to test glucose-induced insulin release and total catecholamine from the adrenal glands was measured. Mice were also submitted to intraperitoneal glucose tolerance test. MSG-obese mice showed fasting hyperglycemia, hyperinsulinemia, and glucose intolerance. Severe reduction of adrenal catecholamines content has also been reported. Besides, the inhibition of fat tissue accretion, exercise caused normalization of insulin blood levels and glycemic control. The pancreatic islets of obese mice, with impaired glucose induced insulin secretion, were recovered after swimming exercises. Adrenal catecholamine content was increased by swimming. Results show that attenuation of MSG-hypothalamic obesity onset is caused, at least in part, by modulation of sympathoadrenal axis activity imposed by early exercise, which may be associated with subsequent glucose metabolism improvement. PMID- 19297407 TI - Tuberous sclerosis complex, implication from a rare genetic disease to common cancer treatment. AB - Tuberous sclerosis complex (TSC) is a relatively rare autosomal dominant disorder characterized by widespread benign tumor formation in a variety of organs. Mutations in either TSC1 or TSC2 tumor suppressor gene are responsible for TSC. The gene products of TSC1 and TSC2, also known as hamartin and tuberin, respectively, form a physical and functional complex and inhibit the mammalian target of rapamycin complex 1 (mTORC1) signaling. The mTORC1 pathway is an evolutionarily conserved growth promoting pathway. mTORC1 plays an essential role in a wide array of cellular processes including translation, transcription, trafficking and autophagy. In this review, we will discuss recent progresses in the TSC-mTOR field and their physiological functions and alterations of this pathway in pathophysiology. PMID- 19297409 TI - Foxg1 promotes olfactory neurogenesis by antagonizing Gdf11. AB - Foxg1, a winged-helix transcription factor, promotes the development of anterior neural structures; in mice lacking Foxg1, development of the cerebral hemispheres and olfactory epithelium (OE) is severely reduced. It has been suggested that Foxg1 acts by positively regulating the expression of growth factors, such as Fgf8, which support neurogenesis. However, Foxg1 also binds Smad transcriptional complexes, allowing it to negatively regulate the effects of TGFbeta family ligands. Here, we provide evidence that this latter effect explains much of the ability of Foxg1 to drive neurogenesis in the OE. We show that Foxg1 is expressed in developing OE at the same time as the gene encoding growth differentiation factor 11 (Gdf11), a TGFbeta family member that mediates negative-feedback control of OE neurogenesis. Mutations in Gdf11 rescue, to a considerable degree, the major defects in Foxg1(-/-) OE, including the early, severe loss of neural precursors and olfactory receptor neurons, and the subsequent collapse of both neurogenesis and nasal cavity formation. Rescue is gene-dosage dependent, with loss of even one allele of Gdf11 restoring substantial neurogenesis. Notably, we find no evidence for a disruption of Fgf8 expression in Foxg1(-/-) OE. However, we do observe both a failure of expression of follistatin (Fst), which encodes a secreted Gdf11 antagonist normally expressed in and around OE, and an increase in the expression of Gdf11 itself within the remaining OE in these mutants. Fst expression is rescued in Foxg1(-/-);Gdf11(-/-) and Foxg1(-/-);Gdf11(+/-) mice. These data suggest that the influence of Foxg1 on Gdf11-mediated negative feedback of neurogenesis may be both direct and indirect. In addition, defects in development of the cerebral hemispheres in Foxg1(-/-) mice are not rescued by mutations in Gdf11, nor is Gdf11 expressed at high levels within these structures. Thus, the pro-neurogenic effects of Foxg1 are likely to be mediated through different signaling pathways in different parts of the nervous system. PMID- 19297410 TI - Common genetic control of haemangioblast and cardiac development in zebrafish. AB - Over the past few years it has become clear that over half of the mammalian heart derives from outside the heart field as originally defined. Such a second heart field, however, has not been described in zebrafish, which could explain its smaller, two-chambered heart. Instead, zebrafish have a population of haemangioblasts, which is absent in mammalian embryos, raising the possibility that these cells represent the evolutionary ancestor of the second heart field. Here, we show for the first time that the genetic programmes of these anterior haemangioblasts and the adjacent heart field are co-regulated, by transcription factors previously associated with heart but not blood or endothelial development. We demonstrate that gata4, gata5 and gata6 are essential for anterior haemangioblast specification, and for subsequent myelopoiesis, acting as early as cloche and upstream of scl. The requirement for gata4, gata5 and gata6 in myeloid, endothelial and cardiac specification is in the mesoderm, but these factors also control, from within the endoderm and the yolk syncytial layer, the migration of the cardiac precursors as they differentiate. This genetic link between the blood/endothelial and cardiac programmes supports the notion that this haemangioblast population in zebrafish is an evolutionary antecedent of the second heart field, and has implications for the differentiation of haemangioblasts and cardiomyocytes from pluripotent cells, and for the origins of stem cells in the adult heart. PMID- 19297412 TI - Drosophila Tubulin-specific chaperone E functions at neuromuscular synapses and is required for microtubule network formation. AB - Hypoparathyroidism, mental retardation and facial dysmorphism (HRD) is a fatal developmental disease caused by mutations in tubulin-specific chaperone E (TBCE). A mouse Tbce mutation causes progressive motor neuronopathy. To dissect the functions of TBCE and the pathogenesis of HRD, we generated mutations in Drosophila tbce, and manipulated its expression in a tissue-specific manner. Drosophila tbce nulls are embryonic lethal. Tissue-specific knockdown and overexpression of tbce in neuromusculature resulted in disrupted and increased microtubules, respectively. Alterations in TBCE expression also affected neuromuscular synapses. Genetic analyses revealed an antagonistic interaction between TBCE and the microtubule-severing protein Spastin. Moreover, treatment of muscles with the microtubule-depolymerizing drug nocodazole implicated TBCE as a tubulin polymerizing protein. Taken together, our results demonstrate that TBCE is required for the normal development and function of neuromuscular synapses and that it promotes microtubule formation. As defective microtubules are implicated in many neurological and developmental diseases, our work on TBCE may offer novel insights into their basis. PMID- 19297411 TI - RacGAP50C directs perinuclear gamma-tubulin localization to organize the uniform microtubule array required for Drosophila myotube extension. AB - The microtubule (MT) cytoskeleton is reorganized during myogenesis as individual myoblasts fuse into multinucleated myotubes. Although this reorganization has long been observed in cell culture, these findings have not been validated during development, and proteins that regulate this process are largely unknown. We have identified a novel postmitotic function for the cytokinesis proteins RacGAP50C (Tumbleweed) and Pavarotti as essential regulators of MT organization during Drosophila myogenesis. We show that the localization of the MT nucleator gamma tubulin changes from diffuse cytoplasmic staining in mononucleated myoblasts to discrete cytoplasmic puncta at the nuclear periphery in multinucleated myoblasts, and that this change in localization depends on RacGAP50C. RacGAP50C and gamma tubulin colocalize at perinuclear sites in myotubes, and in RacGAP50C mutants gamma-tubulin remains dispersed throughout the cytoplasm. Furthermore, we show that the mislocalization of RacGAP50C in pavarotti mutants is sufficient to redistribute gamma-tubulin to the muscle fiber ends. Finally, myotubes in RacGAP50C mutants have MTs with non-uniform polarity, resulting in multiple guidance errors. Taken together, these findings provide strong evidence that the reorganization of the MT network that has been observed in vitro plays an important role in myotube extension and muscle patterning in vivo, and also identify two molecules crucial for this process. PMID- 19297413 TI - C. elegans mig-6 encodes papilin isoforms that affect distinct aspects of DTC migration, and interacts genetically with mig-17 and collagen IV. AB - The gonad arms of C. elegans hermaphrodites acquire invariant shapes by guided migrations of distal tip cells (DTCs), which occur in three phases that differ in the direction and basement membrane substrata used for movement. We found that mig-6 encodes long (MIG-6L) and short (MIG-6S) isoforms of the extracellular matrix protein papilin, each required for distinct aspects of DTC migration. Both MIG-6 isoforms have a predicted N-terminal papilin cassette, lagrin repeats and C terminal Kunitz-type serine proteinase inhibitory domains. We show that mutations affecting MIG-6L specifically and cell-autonomously decrease the rate of post embryonic DTC migration, mimicking a post-embryonic collagen IV deficit. We also show that MIG-6S has two separable functions - one in embryogenesis and one in the second phase of DTC migration. Genetic data suggest that MIG-6S functions in the same pathway as the MIG-17/ADAMTS metalloproteinase for guiding phase 2 DTC migrations, and MIG-17 is abnormally localized in mig-6 class-s mutants. Genetic data also suggest that MIG-6S and non-fibrillar network collagen IV play antagonistic roles to ensure normal phase 2 DTC guidance. PMID- 19297414 TI - Ptch1 is required locally for mammary gland morphogenesis and systemically for ductal elongation. AB - Systemic hormones and local growth factor-mediated tissue interactions are essential for mammary gland development. Using phenotypic and transplantation analyses of mice carrying the mesenchymal dysplasia (mes) allele of patched 1 (Ptch1(mes)), we found that Ptch1(mes) homozygosity led to either complete failure of gland development, failure of post-pubertal ductal elongation, or delayed growth with ductal dysplasia. All ductal phenotypes could be present in the same animal. Whole gland and epithelial fragment transplantation each yielded unique morphological defects indicating both epithelial and stromal functions for Ptch1. However, ductal elongation was rescued in all cases, suggesting an additional systemic function. Epithelial function was confirmed using a conditional null Ptch1 allele via MMTV-Cre-mediated disruption. In Ptch1(mes) homozygotes, failure of ductal elongation correlated with diminished estrogen and progesterone receptor expression, but could not be rescued by exogenous ovarian hormone treatment. By contrast, pituitary isografts were able to rescue the ductal elongation phenotype. Thus, Ptch1 functions in the mammary epithelium and stroma to regulate ductal morphogenesis, and in the pituitary to regulate ductal elongation and ovarian hormone responsiveness. PMID- 19297416 TI - Small effects of treatments for non-specific low back pain: how can we improve patients' outcomes? PMID- 19297417 TI - Do mitochondrial mutations cause recurrent miscarriage? AB - The cause of recurrent miscarriage (RM) can be identified in approximately 50% of cases, whereas in others, unknown genetic factors are actively being sought. As mitochondrial functions, and therefore also the mitochondrial genome [mitochondrial DNA (mtDNA)], have an important role in human development, through ATP production and participation in apoptosis, we aimed to study the role of mtDNA variations in RM. We screened 48 women with RM and 48 age-matched control women for heteroplasmic mitochondrial mutations using denaturing high performance liquid chromatography, a sensitive method that can detect approximately 5% heteroplasmy. As a result, we detected a heteroplasmic mtDNA variation in 13 RM women (27%) and in 9 control women (19%). Seven synonymous and five non synonymous changes were detected within coding regions. In addition, seven heteroplasmic variations were detected within the non-coding control region. We were also able to show the presence of the variations in eight placental samples from three heteroplasmic women. In three of these cases, the proportion of variant mtDNA was higher in the placenta compared with that in the mother. We conclude that our sensitive methodology revealed a higher frequency of samples with heteroplasmic variations than expected in women with both RM and controls. However, no apparent increased frequency of heteroplasmic mtDNA variations or amounts of aberrant mtDNA was detected in the RM group. In addition, none of the detected variations were previously known to be pathogenic and therefore they are an unlikely cause of miscarriage. PMID- 19297418 TI - Comparative methylation profiles and telomerase biology of mouse multipotent adult germline stem cells and embryonic stem cells. AB - Recently, several groups described the isolation of mouse spermatogonial stem cells (SSCs) and their potential to develop to embryonic stem cell (ESC)-like cells, so-called multipotent germline stem cells (mGSCs). We were the first to derive such mGSCs from SSCs isolated from adult mouse testis and, therefore, called these mGSCs multipotent adult germline stem cells (maGSCs). Here, we comparatively analyzed gene-specific and global DNA methylation profiles as well as the telomerase biology of several maGSC and male ESC lines. We show that undifferentiated maGSCs are very similar to undifferentiated male ESCs with regard to global DNA methylation, methylation of pluripotency marker gene loci, telomerase activity and telomere length. Imprinted gene methylation levels were generally lower in undifferentiated maGSCs than in undifferentiated male ESCs, but, compared with undifferentiated mGSCs derived by other groups, more similar to those of male ESCs. Differentiation of maGSCs increased the methylation of three of the four analyzed imprinted genes to almost somatic methylation patterns, but dramatically decreased global DNA methylation. Our findings further substantiate the pluripotency of maGSCs and their potential for regenerative medicine. PMID- 19297419 TI - Altered activity of lysophospholipase D, which produces bioactive lysophosphatidic acid and choline, in serum from women with pathological pregnancy. AB - Altered lipid metabolism is associated with human abnormal pregnancy, such as pre eclampsia and preterm labor, and potentially leads to fetus loss. A causative factor for the onset and progress of the systemic multifactorial syndromes associated with the pathological pregnancy is oxidized low-density lipoprotein, an active identity of which was postulated to be lysophosphatidic acid (LPA). We previously found that LPA is produced extracellularly by plasma lysophospholipase D (lysoPLD) activity of autotaxin, a tumor cell motility-stimulating protein. In this study, a convenient assay based on the choline released from endogenous substrate or exogenous lysophosphatidylcholine (LPC) was used for comparison of serum lysoPLD activity among patients with normal and abnormal pregnancy. The serum choline-producing activity was found to be mainly due to autotaxin, and dependent on its dilution rate. There was some association between low dilution dependency of serum lysoPLD activity toward an exogenous LPC and high lysoPLD activity toward endogenous substrates in cases of patients with preterm labor and pre-eclampsia. However, there was no difference in the serum level of LPC between women with normal pregnancy and those with pathological pregnancy. These results indicate that production of bioactive LPA by lysoPLD activity is elevated by an unknown mechanism that may be related to increased availability of endogenous substrates LPC, but not its concentration in human serum. If the level of LPA in blood circulation is elevated in the pathological pregnancies in vivo, it may play a role in induction and/or progression of systemic vascular dysfunction seen patients with preterm labor or pre-eclampsia. PMID- 19297420 TI - Influence of magnesium ion on the binding of p53 DNA-binding domain to DNA response elements. AB - Site-specific recognition and DNA-binding activity of p53 are crucial for its tumour suppressor function. Previous reports have shown that metal ions can affect the specific recognition and DNA-binding activity of p53DBD. Here we firstly report that magnesium ion can bind to the protein and influence its DNA binding activity. To elucidate the nature and the effect of metal ions in the reaction chemistry, we utilized endogenous tryptophan fluorescence to quantitate the interaction between p53DBD and metal ions. The K(a) value for the binding of Mg(2+) to the protein is 1.88 x 10(3) M(-1). Analysis of the CD data clearly suggested that the binding of magnesium ion induced a subtle conformational change rather than a radical modification of the overall protein architecture. Based on the results of electrophoretic mobility shift assays and fluorescence experiments, we concluded that the binding of Mg(2+) significantly stimulated the binding of the protein to DNA in a sequence-independent manner, which differed from that of zinc ions in a sequence-specific manner. Based on these results and the fact that Mg(2+) exists at relatively high concentration in the cell, we propose that Mg(2+) is one of potential factors to affect or regulate the transactivation of p53. PMID- 19297421 TI - An orally bioavailable synthetic analog of an active dehydroepiandrosterone metabolite reduces established disease in rodent models of rheumatoid arthritis. AB - Dehydroepiandrosterone (DHEA) treatment provides diverse anti-inflammatory benefits in rodent models of diseases, including rheumatoid arthritis (RA), but only limited benefits to patients. In rodents, DHEA is metabolized to (among others) androstene-3beta,7beta,17beta-triol (AET), which retains potent anti inflammatory activity. 17Alpha-ethynyl-5-androstene-3beta,7beta,17beta-triol (HE3286) is a novel, metabolically stabilized, orally bioavailable derivative of AET. In the DBA mouse model of collagen-induced arthritis (CIA), once-daily oral treatments (gavage) with HE3286 (40 mg/kg), beginning at onset of disease, significantly decreased disease. Benefit was associated with reduction in joint inflammation, erosion, and synovial proliferation as measured by histological analysis and mRNA of proinflammatory cytokines, including tumor necrosis factor alpha, interleukin (IL)-6, IL-1beta, and IL-23. Significant benefit was also observed in the CIA model even when treatments were delayed until 7 days after the onset of arthritis. Furthermore, dose-dependent benefit was observed in the DBA mouse model of collagen antibody-induced arthritis, as well as reductions in IL-6 and matrix metalloproteinase-3 mRNA levels in joints at the peak of disease and at the end of the study. HE3286, in contrast to dexamethasone, was not immune suppressive in several classic animal models of immune function. Instead, HE3286 treatment was associated with reduced nuclear factor-kappaB activation and in our previous studies, with increased regulatory T cells. We hypothesize that HE3286 may represent a novel, perhaps first-in-class, anti-inflammatory agent and may more fully translate the benefits of DHEA, heretofore largely limited to rodents, into treatments for human diseases, including autoimmune disorders such as RA. PMID- 19297422 TI - Endothelin ET(B) receptors in arteries and veins: multiple actions in the vein. AB - Endothelin receptors (ET(A) and ET(B)) mediate responses to ET-1. ET(B) receptor function seems to differ between a similarly sized arterial and venous pair, the rat vena cava (RVC) and rat thoracic aorta (RA). ET(B) receptors mediate RVC contraction directly, but it is unclear whether ET(B) receptors mediate contraction in RA. Because of these apparent differences in ET(B) receptor mediated vascular contraction, we hypothesize that relaxant ET(B)-receptor mechanisms in RVC would be different from those in RA. RA and RVC rings were isolated from rats for measurement of isometric contraction. When contracted with prostaglandin F-2alpha (PGF-2alpha) (20 microM), the ET(B) receptor agonist sarafotoxin-6c (S6c) (100 nM) significantly relaxed RA and RVC. N(omega)-Nitro-L arginine (LNNA) (100 microM) or endothelial denudation abolished relaxation to S6c in RA. By contrast, S6c-induced relaxation of RVC was attenuated but not abolished by LNNA and endothelial denudation. RVC (PGF-2alpha-contracted) relaxed to low concentrations of ET-1, whereas under the same conditions RA responded with contraction. ET-1-induced relaxation in RA was observed only with ET(A) receptor blockade. Vessels from dopamine-beta-hydroxylase-ET(B) transgenic rats, which lack functional ET(B) receptors in the vasculature, were also used. RVC (PGF-2alpha-contracted) from these rats did not relax to ET-1. Thus, although both RA and RVC possess endothelial relaxant ET(B) receptors, RA and RVC differ in that relaxant ET(B) receptors may also be present in smooth muscle of RVC. Moreover, the mechanisms of endothelial cell ET(B) receptor-mediated relaxation in RA and RVC are not the same. PMID- 19297423 TI - Curcumin inhibits adipogenesis in 3T3-L1 adipocytes and angiogenesis and obesity in C57/BL mice. AB - Angiogenesis is necessary for the growth of adipose tissue. Dietary polyphenols may suppress growth of adipose tissue through their antiangiogenic activity and by modulating adipocyte metabolism. We investigated the effect of curcumin, the major polyphenol in turmeric spice, on angiogenesis, adipogenesis, differentiation, apoptosis, and gene expression involved in lipid and energy metabolism in 3T3-L1 adipocyte in cell culture systems and on body weight gain and adiposity in mice fed a high-fat diet (22%) supplemented with 500 mg curcumin/kg diet for 12 wk. Curcumin (5-20 micromol/L) suppressed 3T3-L1 differentiation, caused apoptosis, and inhibited adipokine-induced angiogenesis of human umbilical vein endothelial cells. Supplementing the high-fat diet of mice with curcumin did not affect food intake but reduced body weight gain, adiposity, and microvessel density in adipose tissue, which coincided with reduced expression of vascular endothelial growth factor (VEGF) and its receptor VEGFR-2. Curcumin increased 5'AMP-activated protein kinase phosphorylation, reduced glycerol-3-phosphate acyl transferase-1, and increased carnitine palmitoyltransferase-1 expression, which led to increased oxidation and decreased fatty acid esterification. The in vivo effect of curcumin on the expression of these enzymes was also confirmed by real-time RT-PCR in subcutaneous adipose tissue. In addition, curcumin significantly lowered serum cholesterol and expression of PPARgamma and CCAAT/enhancer binding protein alpha, 2 key transcription factors in adipogenesis and lipogenesis. The curcumin suppression of angiogenesis in adipose tissue together with its effect on lipid metabolism in adipocytes may contribute to lower body fat and body weight gain. Our findings suggest that dietary curcumin may have a potential benefit in preventing obesity. PMID- 19297424 TI - Gamma-tocotrienol and gamma-tocopherol are primarily metabolized to conjugated 2 (beta-carboxyethyl)-6-hydroxy-2,7,8-trimethylchroman and sulfated long-chain carboxychromanols in rats. AB - The metabolism of gamma-tocotrienol (gamma-TE) and gamma-tocopherol (gamma-T) was investigated in human A549 cells and in rats. Similar to gamma-T, A549 cells metabolized gamma-TE to sulfated 9'-, 11'-, and 13'-carboxychromanol and their unconjugated counterparts. After 72-h incubation with the cells, 90% of long chain carboxychromanols in the culture media from gamma-TE, but <45% from gamma T, were in the sulfated form. The formation of these metabolites was further investigated in rats gavaged by gamma-TE at 10 or 50 mg/kg, gamma-T at 10 mg/kg, or tocopherol-stripped corn oil in controls. Six hours after a single dosing, the supplemented rats had increased plasma concentrations of 13'-carboxychromanol and sulfated 9'-, 11'-, 13'-carboxychromanol, whereas none of these metabolites were detectable in the controls. Sulfated 11'-carboxychromanol was the most abundant long-chain metabolite in gamma-TE-supplemented rats. Sulfatase/glucuronidase hydrolysis revealed for the first time that >88% 2-(beta-carboxyethyl)-6 hydroxychroman (gamma-CEHC), the terminal beta-oxidation metabolite, was in the conjugated form in the plasma. In all groups, conjugated gamma-CEHC accounted for >75% of total metabolites, whereas free CEHC was a minor metabolite. At 10 mg/kg, the plasma concentrations of total metabolites from gamma-TE-supplemented rats were higher (P < 0.05) than those from gamma-T-fed rats. These results demonstrate that in rats, conjugation such as sulfation occurs parallel to beta oxidation in the liver and is quantitatively important to vitamin E metabolism. Conjugated long-chain carboxychromanols may be novel excreted metabolites during supplementation. Our data also provide in vivo evidence that gamma-TE is more extensively metabolized than gamma-T. PMID- 19297425 TI - Elevated corticosterone associated with food deprivation upregulates expression in rat skeletal muscle of the mTORC1 repressor, REDD1. AB - Food deprivation induces a repression of protein synthesis in skeletal muscle in part due to reduced signaling through the mammalian target of rapamycin complex 1 (mTORC1). Previous studies have identified upregulated expression of the protein Regulated in DNA Damage and Development (REDD1) as an important mechanism in the regulation of mTORC1 activity in response to a variety of stresses. Our goal in this investigation was to determine whether modulation of REDD1 expression occurs in response to food deprivation and refeeding, and, if it does, to ascertain if changes in REDD1 expression correlate with altered mTORC1 signaling. As expected, mTORC1 signaling was repressed after 18 h of food deprivation compared with freely-fed control rats and quickly recovered after refeeding for 45 min. Food deprivation caused a dramatic rise in REDD1 mRNA and protein expression; refeeding resulted in a reduction to baseline. Food deprivation is characterized by low-serum insulin and elevated glucocorticoid concentrations. Therefore, initially, alloxan-induced type I diabetes was used to minimize the food deprivation- and refeeding-induced changes in insulin. Although diabetic rats exhibited upregulated REDD1 expression compared with nondiabetic controls, there was no direct correlation between REDD1 mRNA expression and serum insulin levels, and insulin treatment of diabetic rats did not affect REDD1 expression. In contrast, serum corticosterone levels correlated directly with REDD1 mRNA expression (r = 0.68; P = 0.01). Moreover, inhibiting corticosterone-mediated signaling via administration of the glucocorticoid receptor antagonist RU486 blocked both the food deprivation- and diabetes-induced increase in REDD1 mRNA expression. Overall, the results demonstrate that changes in REDD1 expression likely contribute to the regulation of mTORC1 signaling during food deprivation and refeeding. PMID- 19297426 TI - Digestive stability of xanthophylls exceeds that of carotenes as studied in a dynamic in vitro gastrointestinal system. AB - Epidemiological studies have suggested that high consumption of tomato products is associated with a lower risk for chronic diseases. To exert their health effect, the phytochemicals of tomatoes have to be bioavailable and therefore it implies their stability through the digestion process. Here, we assessed the digestive stability of the red-pigmented lycopene and other carotenoids brought in nutritional quantity within different food matrixes, using the TNO gastrointestinal tract model (TIM). This multicompartmental dynamic system accurately reproduces the main parameters of gastric and small intestinal digestion in human. In vitro digestions of a standard meal containing red tomato (RT), yellow tomato (devoid of lycopene), or lycopene beadlets were performed. Zeaxanthin and lutein were stable throughout artificial digestions, whereas beta carotene and all-trans lycopene were degraded (approximately 30 and 20% loss at the end of digestion, respectively) in the jejunal and ileal compartments. The recovery of beta-carotene in the digesta of the RT meal was significantly lower than that in the yellow one, showing a food matrix effect. In the same way, until 180 min of digestion, the recovery percentages of all-trans lycopene from RT were significantly lower than those issued from the supplement. Isomeric conformation also influenced the stability of carotenoids, 5-cis lycopene being the most stable isomer followed by all-trans and 9-cis. No trans-cis isomerization of lycopene occurred in the TIM. By using a relevant dynamic in vitro system, this study allowed us to gain further insight into the parameters influencing the digestive stability of carotenoids, and therefore their bioavailability, in humans. PMID- 19297427 TI - Tissue-specific alterations in zinc transporter expression in intestine and liver reflect a threshold for homeostatic compensation during dietary zinc deficiency in weanling rats. AB - Intestinal zinc (Zn) absorption and liver Zn mobilization are presumed to regulate Zn homeostasis. Several Zn transporters have been identified; however, their contribution to Zn homeostasis is poorly understood. Moreover, their regulation during periods of growth is unknown. To characterize the mechanisms that maintain Zn status, weanling rats were fed control (25 mg/kg), marginally low (MLZ; 15 mg/kg), low (LZ; 7 mg/kg), or very low (VLZ; <1 mg/kg) Zn diets for 3 wk and effects on jejunum Zip4 and ZnT1 and hepatic Zip1 and ZnT1 were assessed. Another control group was pair-fed (PF) to VLZ. The MLZ rats had lower jejunum ZnT1 protein abundance than the control. In the LZ group, we detected increased jejunum Zip4 mRNA expression and hepatic ZnT1 protein abundance and reduced jejunum Zip4 and ZnT1 and hepatic Zip1 protein abundance. VLZ had lower jejunum ZnT1 mRNA and protein abundance and hepatic Zip1 and ZnT1 protein abundance compared with the PF group. Zip4 protein was present at the intestinal villus tip in controls but was detected on the apical membrane throughout the entire villus in LZ rats. ZnT5 protein in jejunum was always detected at the apical membrane and also at the basolateral membrane of VLZ rats. In contrast, ZnT7 was found intracellularly in jejunum. Our data suggest that effects of Zn deficiency on Zn homeostasis occurs biphasically during marginal Zn deficiency through increased intestinal Zn uptake capacity and reduced intestinal Zn efflux, then during more pronounced degrees of Zn deficiency through decreased liver Zn accretion and increased hepatic Zn efflux back into circulation. These results assist in our understanding of how mammals regulate Zn homeostasis. PMID- 19297429 TI - Quercetin enhances intestinal barrier function through the assembly of zonula [corrected] occludens-2, occludin, and claudin-1 and the expression of claudin-4 in Caco-2 cells. AB - Dietary flavonoids provide various beneficial effects for our health. We investigated the promotive effects of quercetin and myricetin on the intestinal barrier function in human intestinal Caco-2 cell monolayers. Transepithelial electrical resistance (TER) across the monolayers increased rapidly during incubation with quercetin, peaking at 6 h. Lucifer yellow flux, a paracellular marker, was dose-dependently lower after quercetin and myricetin treatments, although quercetin exhibited a more potent effect. Immunoblot analysis of tight junction (TJ) proteins revealed that zonula occludens (ZO)-2, occludin, and claudin-1 were distributed to the actin cytoskeleton fraction by quercetin without increasing their respective whole-cell levels and this distribution was correlated with the increases in TER. The claudin-4 level was elevated by quercetin in both the cytoskeleton fraction and whole cells after 12 h. Confocal microscopy showed the assembly of claudin-1 and -4 at the TJ by quercetin. An inhibitor of protein kinase Cdelta (PKCdelta), rottlerin, enhanced the barrier function with changes in the distribution and expression of TJ proteins in a manner very similar to that of quercetin. Phosphorylation of PKCdelta indicating the enzymatic activity in the cells was decreased by quercetin after 1 h. In the kinase assay, quercetin exhibits direct inhibition of the PKCdelta isoform. This study demonstrates that quercetin enhances the intestinal barrier function through the assembly of ZO-2, occludin, and claudin-1 by inhibiting PKCdelta and the increase in claudin-4 expression has an additional role after 12 h. PMID- 19297430 TI - Baseline serum C-reactive protein is associated with lipid responses to low-fat and high-polyunsaturated fat diets. AB - Baseline serum C-reactive protein (CRP) concentrations play a role in the lipid response to diet. This study was a randomized, cross-over, controlled feeding study with 3 phases of 25 d each aimed at determining whether baseline CRP concentrations modulate the serum lipid response to diets differing in fat type and quantity. Participants were adult men and women, age 19-65 y, with LDL cholesterol concentrations of 3.37-4.66 mmol/L. All participants consumed 3 diets differing in the type of snack, either low or high in fat: low-fat (30.8% of energy), moderate in fat and saturated fat (37.9 and 11.4% of energy, respectively), or moderate in fat and polyunsaturated fat (36.3 and 9.7% of energy, respectively). Using baseline CRP as a continuous variable, CRP x diet interactions on change in serum lipoprotein_a (P = 0.045) and HDL-cholesterol (P = 0.06) were observed. When we used previously established categories to define CRP concentrations (low, <1 mg/L; intermediate, 1-3 mg/L; or high, >3 mg/L), we found a CRP x diet interaction on change in triglyceride concentrations (P = 0.03) and trends for CRP x diet interaction on change in LDL (P = 0.06) and total cholesterol (P = 0.07). If replicated, these results suggest that considering baseline CRP concentrations when prescribing dietary interventions to lower lipid concentrations may be useful. Individuals with high baseline CRP concentrations may benefit from moderate-fat, high polyunsaturated fat diets, whereas those with low baseline CRP concentrations may obtain greater lipid-lowering benefits from low-fat diets. PMID- 19297431 TI - Reducing the risk of injury in young footballers. PMID- 19297428 TI - Human CYP3A4 and murine Cyp3A11 are regulated by equol and genistein via the pregnane X receptor in a species-specific manner. AB - Pregnane X receptor (PXR) is an important component of the body's adaptive defense system responsible for the elimination of various toxic xenobiotics. PXR activation by endogenous and exogenous chemicals, including steroids, antibiotics, bile acids, and herbal compounds, results in induction of drug metabolism. We investigated the ability of the isoflavones genistein, daidzein, and the daidzein metabolite equol to activate human and mouse PXR in vitro using cell-based transient transfection studies and primary hepatocytes and in vivo in a mouse model. In transient transfection assays, the isoflavones genistein and daidzein activate full-length, wild-type mouse PXR, but not a mutant form, with genistein being the most potent. In contrast, equol was a more potent activator of human PXR than genistein or daidzein. In a mammalian 2-hybrid assay, isoflavones induced recruitment of the coactivator steroid receptor coactivator 1 to PXR. When tested against the native human Cytochrome P450 3A4 (CYP3A4) promoter, equol was the more potent activator and treatment of human hepatocytes with equol increased CYP3A4 mRNA and immunoreactive protein expression. Treatment of wild-type, but not PXR(-/-), mouse hepatocytes showed that genistein and daidzein induced the expression of Cytochrome P450 3A11 (Cyp3A11) mRNA, whereas equol had no effect. Cyp3A11 mRNA was also induced in vivo in mice fed a soy protein-containing diet. The results presented herein demonstrate that there is a species-specific difference in the activation of PXR by isoflavones and equol. PMID- 19297437 TI - World drugs problem resists all attempts to curb it, says EU report. PMID- 19297441 TI - Reneging on funding commitments could leave health systems in poorer countries $30bn short by 2015. PMID- 19297443 TI - Danish night shift workers with breast cancer awarded compensation. PMID- 19297444 TI - Standardised consultations in primary care. PMID- 19297445 TI - Patient information sheets in emergency care. PMID- 19297446 TI - Measuring the quality of hospital care. PMID- 19297447 TI - Evidence of methodological bias in hospital standardised mortality ratios: retrospective database study of English hospitals. AB - OBJECTIVE: To assess the validity of case mix adjustment methods used to derive standardised mortality ratios for hospitals, by examining the consistency of relations between risk factors and mortality across hospitals. DESIGN: Retrospective analysis of routinely collected hospital data comparing observed deaths with deaths predicted by the Dr Foster Unit case mix method. SETTING: Four acute National Health Service hospitals in the West Midlands (England) with case mix adjusted standardised mortality ratios ranging from 88 to 140. PARTICIPANTS: 96 948 (April 2005 to March 2006), 126 695 (April 2006 to March 2007), and 62 639 (April to October 2007) admissions to the four hospitals. MAIN OUTCOME MEASURES: Presence of large interaction effects between case mix variable and hospital in a logistic regression model indicating non-constant risk relations, and plausible mechanisms that could give rise to these effects. RESULTS: Large significant (P 1/4). These results indicate that AQP4 organization and expression are regulated by vasopressin in vivo and in vitro and demonstrate that the dominant role for M1 is restricted to a one-to-one interaction between AQP4 splice variants that regulates the membrane expression of OAPs. PMID- 19297455 TI - Alcohol-related and hepatocellular cancer deaths by country of birth in England and Wales: analysis of mortality and census data. AB - BACKGROUND: The incidence of and mortality from alcohol-related conditions, liver disease and hepatocellular cancer (HCC) are increasing in the UK. We compared mortality rates by country of birth to explore potential inequalities and inform clinical and preventive care. DESIGN: Analysis of mortality for people aged 20 years and over using the 2001 Census data and death data from 1999 and 2001-2003. SETTING: England and Wales. MAIN OUTCOME MEASURES: Standardized mortality ratios (SMRs) for alcohol-related deaths and HCC. RESULTS: Mortality from alcohol related deaths (23 502 deaths) was particularly high for people born in Ireland (SMR for men [M]: 236, 95% confidence interval [CI]: 219-254; SMR for women [F]: 212, 95% CI: 191-235) and Scotland (SMR-M: 187, CI: 173-213; SMR-F 182, CI: 163 205) and men born in India (SMR-M: 161, CI: 144-181). Low alcohol-related mortality was found in women born in other countries and men born in Bangladesh, Middle East, West Africa, Pakistan, China and Hong Kong, and the West Indies. Similar mortality patterns were observed by country of birth for alcoholic liver disease and other liver diseases. Mortality from HCC (8266 deaths) was particularly high for people born in Bangladesh (SMR-M: 523, CI: 380-701; SMR-F: 319, CI: 146-605), China and Hong Kong (SMR-M: 492, CI: 168-667; SMR-F: 323, CI: 184-524), West Africa (SMR-M: 440, CI, 308-609; SMR-F: 319, CI: 165-557) and Pakistan (SMR-M: 216, CI: 113-287; SMR-F: 215, CI: 133-319). CONCLUSIONS: These findings show persistent differences in mortality by country of birth for both alcohol-related and HCC deaths and have important clinical and public health implications. New policy, research and practical action are required to address these differences. PMID- 19297456 TI - Sex differences in inflammatory markers: what is the contribution of visceral adiposity? AB - BACKGROUND: C-reactive protein (CRP) concentrations have been found to be higher in premenopausal women than in men, whereas interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) concentrations have been reported to be lower in women than in men. OBJECTIVE: The objective was to determine whether the sex difference in body fat distribution accounts for the observed sex differences in inflammatory markers. DESIGN: Plasma CRP, IL-6, and TNF-alpha concentrations were measured in 208 healthy men (age: 42.2 +/- 15.2 y) and in 145 healthy women (age: 36.8 +/- 11.1 y). RESULTS: Compared with men, premenopausal women had higher CRP concentrations [1.24 (25th percentile: 0.54; 75th percentile: 3.04) compared with 0.94 (0.51, 2.40) mg/L; P < 0.05] and lower plasma TNF-alpha concentrations [1.50 (25th percentile: 1.23; 75th percentile: 1.82) compared with 1.71 (1.40, 2.05) pg/mL; P < 0.001]. No sex difference in IL-6 concentrations was noted. Regression analyses indicated that the relation between CRP or IL-6 and visceral adipose tissue (VAT) and subcutaneous AT (SAT) was sex-specific; a significantly steeper slope was observed in women than in men (P < 0.05). Sex differences in CRP concentrations were abolished after SAT was adjusted for. In a multivariate model of the whole sample, we found that both SAT and VAT and the sex x SAT interaction term were significant correlates of CRP and IL-6 concentrations. Finally, whereas CRP concentrations were largely influenced by visceral adiposity in men, subcutaneous adiposity was the key correlate of CRP in women. CONCLUSION: The higher CRP concentrations found in women appear to be due to their greater accumulation of subcutaneous fat than that observed in men. PMID- 19297458 TI - Mortality in British vegetarians: results from the European Prospective Investigation into Cancer and Nutrition (EPIC-Oxford). AB - BACKGROUND: Few prospective studies have examined the mortality of vegetarians. OBJECTIVE: We present results on mortality among vegetarians and nonvegetarians in the European Prospective Investigation into Cancer and Nutrition (EPIC Oxford). DESIGN: We used a prospective study of men and women recruited throughout the United Kingdom in the 1990s. RESULTS: Among 64,234 participants aged 20-89 y for whom diet group was known, 2965 had died before age 90 by 30 June 2007. The death rates of participants are much lower than average for the United Kingdom. The standardized mortality ratio for all causes of death was 52% (95% CI: 50%, 54%) and was identical in vegetarians and in nonvegetarians. Comparing vegetarians with meat eaters among the 47,254 participants who had no prevalent cardiovascular disease or malignant cancer at recruitment, the death rate ratios adjusted for age, sex, smoking, and alcohol consumption were 0.81 (95% CI: 0.57, 1.16) for ischemic heart disease and 1.03 (95% CI: 0.90, 1.16) for all causes of death. CONCLUSIONS: The mortality of both the vegetarians and the nonvegetarians in this study is low compared with national rates. Within the study, mortality from circulatory diseases and all causes is not significantly different between vegetarians and meat eaters, but the study is not large enough to exclude small or moderate differences for specific causes of death, and more research on this topic is required. PMID- 19297457 TI - Size at birth, weight gain in infancy and childhood, and adult blood pressure in 5 low- and middle-income-country cohorts: when does weight gain matter? AB - BACKGROUND: Promoting catch-up growth in malnourished children has health benefits, but recent evidence suggests that accelerated child weight gain increases adult chronic disease risk. OBJECTIVE: We aimed to determine how birth weight (BW) and weight gain to midchildhood relate to blood pressure (BP) in young adults. DESIGN: We pooled data from birth cohorts in Brazil, Guatemala, India, the Philippines, and South Africa. We used conditional weight (CW), a residual of current weight regressed on prior weights, to represent deviations from expected weight gain from 0 to 12, 12 to 24, 24 to 48 mo, and 48 mo to adulthood. Adult BP and risk of prehypertension or hypertension (P/HTN) were modeled before and after adjustment for adult body mass index (BMI) and height. Interactions of CWs with small size-for-gestational age (SGA) at birth were tested. RESULTS: Higher CWs were associated with increased BP and odds of P/HTN, with coefficients proportional to the contribution of each CW to adult BMI. Adjusted for adult height and BMI, no child CW was associated with adult BP, but 1 SD of BW was related to a 0.5-mm Hg lower systolic BP and a 9% lower odds of P/HTN. BW and CW associations with systolic BP and P/HTN were not different between adults born SGA and those with normal BW, but higher CW at 48 mo was associated with higher diastolic BP in those born SGA. CONCLUSIONS: Greater weight gain at any age relates to elevated adult BP, but faster weight gains in infancy and young childhood do not pose a higher risk than do gains at other ages. PMID- 19297459 TI - Early determinants of cardiovascular disease: the role of early diet in later blood pressure control. AB - It is now widely accepted that a gross change in the maternal diet during pregnancy results in offspring with raised blood pressure. More recently, results from human intervention studies and a range of animal experiments have questioned this concept. It thus appears that, when blood pressure is measured directly or by telemetry, the extent to which blood pressure is raised is largely dependent on the magnitude of the postnatal catch-up growth. In addition, such effects can be lost when appropriate corrections are made for current body weight. Consequently, offspring born to nutritionally manipulated mothers can actually have a lower blood pressure than control group offspring. At the same time, studies of the offspring born to contemporary women in developed countries show very little, if any, effect of changes in maternal diet on blood pressure in the offspring when assessed during childhood. In small animal studies, at least, the cardiovascular outcomes linked to small size at birth can differ between the sexes, which may be related in part to differences in kidney function between males and females. With respect to large animal studies, significant effects on blood pressure are less apparent and may relate to the much slower onset of hypertension. The challenge is to use our increased knowledge of the critical windows in early development to optimize later health. One clear priority is the prevention of excess adiposity and to determine how epigenetic mechanisms may provide novel strategies in this regard. PMID- 19297460 TI - Prevalence and predictors of iron deficiency in fully breastfed infants at 6 mo of age: comparison of data from 6 studies. AB - BACKGROUND: Iron deficiency (ID) can occur among exclusively breastfed infants before 6 mo of age. OBJECTIVE: The objective was to determine which subgroups of fully breastfed infants are at highest risk of ID. DESIGN: We assessed the prevalence of ID (ferritin < 12 mug/L) and iron deficiency anemia (IDA; ferritin < 12 mug/L and hemoglobin < 105 g/L) and risk factors associated with ID and IDA at 6 mo among 404 fully breastfed infants with a birth weight >2500 g from 6 studies in Ghana, Honduras, Mexico, and Sweden. Infants with an elevated C reactive protein concentration (8%) were excluded. RESULTS: The percentages of infants with ID were 6% in Sweden, 17% in Mexico, 13-25% in Honduras, and 12-37% in Ghana. The percentages with IDA were 2% in Sweden, 4% in Mexico, 5-11% in Honduras, and 8-16% in Ghana. With data pooled, the key predictors of ID (20%) were male sex [adjusted odds ratio (AOR): 4.6; 95% CI: 2.5, 8.5] and birth weight 2500-2999 g (AOR: 2.4; 95% CI: 1.4, 4.3). The predictors of IDA (8%) were male sex (AOR: 7.6; 95% CI: 2.5, 23.0), birth weight of 2500-2999 g (AOR: 3.4; 1.5, 7.5), and weight gain above the median since birth (AOR: 3.4; 95% CI: 1.3, 8.6). The combination of birth weight 2500-2999 g or male sex had a sensitivity of 91% for identifying ID and of 97% for identifying IDA. CONCLUSIONS: Among fully breastfed infants with a birth weight >2500 g, IDA is uncommon before 6 mo, but male infants and those with a birth weight of 2500-2999 g are at higher risk of ID and IDA. PMID- 19297461 TI - Interindividual differences in response to plant-based diets: implications for cancer risk. AB - Genetic differences in taste preference, food tolerance, and phytochemical absorption and metabolism all potentially influence the effect of plant-based diets on cancer risk. Diet is a mixture of carcinogens, mutagens, and protective agents, many of which are metabolized by biotransformation enzymes. Genetic polymorphisms that alter protein expression or enzyme function can modify risk. Genotypes associated with more favorable handling of carcinogens may be associated with less favorable handling of phytochemicals. For example, glutathione S-transferases detoxify polycyclic aromatic hydrocarbons and metabolize isothiocyanates, which are chemopreventive compounds in cruciferous vegetables. A polymorphism in the GSTM1 gene results in lack of GSTM1-1 protein. Pharmacokinetic studies suggest that lack of GSTM1 enzyme is associated with more rapid excretion of the isothiocyanate sulforaphane; therefore, individuals who have this genetic variation may derive less benefit from consuming cruciferous vegetables. Flavonoids are conjugated with glucuronide and sulfate and are excreted in urine and bile. Polymorphisms in UDP-glucuronosyltransferases and sulfotransferases may contribute to variability in phytochemical clearance and efficacy. Genetic polymorphisms in enzymes that metabolize phytochemicals may account in part for variation in disease risk and also have to be considered in the context of other aspects of human genetics, gut bacterial genetics, and environmental exposures. PMID- 19297462 TI - Estimation of the dietary requirement for vitamin D in free-living adults >=64 y of age. AB - BACKGROUND: Older adults may be more prone to developing vitamin D deficiency than younger adults. Dietary requirements for vitamin D in older adults are based on limited evidence. OBJECTIVE: The objective was to establish the dietary intake of vitamin D required to maintain serum 25-hydroxyvitamin D [25(OH)D] concentrations above various cutoffs between 25 and 80 nmol/L during wintertime, which accounted for the effect of summer sunshine exposure and diet. DESIGN: A randomized, placebo-controlled, double-blind, 22-wk intervention was conducted in men and women aged >/=64 y (n = 225) at supplemental levels of 0, 5, 10, and 15 microg vitamin D(3)/d from October 2007 to March 2008. RESULTS: Clear dose related increments (P < 0.0001) in serum 25(OH)D were observed with increasing supplemental vitamin D(3) intakes. The slope of the relation between total vitamin D intake and serum 25(OH)D was 1.97 nmol . L(-1) . microg intake(-1). The vitamin D intake that maintained serum 25(OH)D concentrations >25 nmol/L in 97.5% of the sample was 8.6 microg/d. Intakes were 7.9 and 11.4 microg/d in those who reported a minimum of 15 min daily summer sunshine exposure or less, respectively. The intakes required to maintain serum 25(OH)D concentrations of >37.5, >50, and >80 nmol/L in 97.5% of the sample were 17.2, 24.7, and 38.7 microg/d, respectively. CONCLUSION: To ensure that the vitamin D requirement is met by the vast majority (>97.5%) of adults aged >/=64 y during winter, between 7.9 and 42.8 microg vitamin D/d is required, depending on summer sun exposure and the threshold of adequacy of 25(OH)D. This trial was registered at http://www.controlled-trials.com/ISRCTN20236112 as ISRCTN registration no. ISRCTN20236112. PMID- 19297463 TI - Food, plant food, and vegetarian diets in the US dietary guidelines: conclusions of an expert panel. AB - We summarize conclusions drawn from a panel discussion at the "Fifth International Congress on Vegetarian Nutrition" about the roles of and emphasis on food, plant food, and vegetarianism in current and future US dietary guidelines. The most general recommendation of the panel was that future dietary guidelines, following the lead of the 2005 Dietary Guidelines Advisory Committee, should emphasize food-based recommendations and thinking to the full extent that evidence allows. Although nutrient-based thinking and Dietary Reference Intakes (DRIs) may help ensure an adequate diet in the sense that deficiency states are avoided, the emphasis on DRIs may not capture many important nutritional issues and may inhibit a focus on foods. More generally, in the context of the conference on vegetarian nutrition, this report focuses on the history and structure of the Dietary Guidelines Advisory Committee, on various plant food oriented recommendations that are supported by literature evidence, and on mechanisms for participating in the process of forming dietary guidelines. Among recommendations that likely would improve health and the environment, some are oriented toward increased plant food consumption and some toward vegetarianism. The literature on health effects of individual foods and whole lifestyle diets is insufficient and justifies a call for future food-oriented research, including expanding the evidence base for plant-based and vegetarian diets. The Dietary Guidelines Advisory Committee's role should be carried forward to creation of a publicly accessible icon (eg, the current pyramid) and related materials to ensure that the science base is fully translated for the public. PMID- 19297464 TI - Role of N-linked glycosylation for sindbis virus infection and replication in vertebrate and invertebrate systems. AB - Each Sindbis virus (SINV) surface glycoprotein has two sites for N-linked glycosylation (E1 positions 139 and 245 [E1-139 and E1-245] and E2 positions 196 and 318 [E2-196 and E2-318]). Studies of SINV strain TE12 mutants with each site eliminated identified the locations of carbohydrates by cryo-electron microscopy (S. V. Pletnev et al., Cell 105:127-136, 2001). In the current study, the effects of altered glycosylation on virion infectivity, growth in cells of vertebrates and invertebrates, heparin binding, virulence in mice, and replication in mosquitoes were assessed. Particle-to-PFU ratios for E1-139 and E2-196 mutant strains were similar to that for TE12, but this ratio for the E1-245 mutant was 100-fold lower than that for TE12. Elimination of either E2 glycosylation site increased virus binding to heparin and increased replication in BHK cells. Elimination of either E1 glycosylation site had no effect on heparin binding but resulted in an approximately 10-fold decrease in virus yield from BHK cells compared to the TE12 amount. No differences in pE2 processing were detected. E2 196 and E2-318 mutants were more virulent in mice after intracerebral inoculation, while E1-139 and E1-245 mutants were less virulent. The E1-245 mutant showed impaired replication in C7/10 mosquito cells and in Culex quinquefasciatus after intrathoracic inoculation. We conclude that the increased replication and virulence of E2-196 and E2-318 mutants are primarily due to increased efficiency of binding to heparan sulfate on mammalian cells. Lack of glycosylation at E1-139 or E1-245 impairs replication in vertebrate cells, while E1-245 also severely affects replication in invertebrate cells. PMID- 19297465 TI - The respiratory syncytial virus matrix protein possesses a Crm1-mediated nuclear export mechanism. AB - The respiratory syncytial virus (RSV) matrix (M) protein is localized in the nucleus of infected cells early in infection but is mostly cytoplasmic late in infection. We have previously shown that M localizes in the nucleus through the action of the importin beta1 nuclear import receptor. Here, we establish for the first time that M's ability to shuttle to the cytoplasm is due to the action of the nuclear export receptor Crm1, as shown in infected cells, and in cells transfected to express green fluorescent protein (GFP)-M fusion proteins. Specific inhibition of Crm1-mediated nuclear export by leptomycin B increased M nuclear accumulation. Analysis of truncated and point-mutated M derivatives indicated that Crm1-dependent nuclear export of M is attributable to a nuclear export signal (NES) within residues 194 to 206. Importantly, inhibition of M nuclear export resulted in reduced virus production, and a recombinant RSV carrying a mutated NES could not be rescued by reverse genetics. That this is likely to be due to the inability of a nuclear export deficient M to localize to regions of virus assembly is indicated by the fact that a nuclear-export deficient GFP-M fails to localize to regions of virus assembly when expressed in cells infected with wild-type RSV. Together, our data suggest that Crm1-dependent nuclear export of M is central to RSV infection, representing the first report of such a mechanism for a paramyxovirus M protein and with important implications for related paramyxoviruses. PMID- 19297466 TI - Type I and type II interferons inhibit the translation of murine norovirus proteins. AB - Human noroviruses are responsible for more than 95% of nonbacterial epidemic gastroenteritis worldwide. Both onset and resolution of disease symptoms are rapid, suggesting that components of the innate immune response are critical in norovirus control. While the study of the human noroviruses has been hampered by the lack of small animal and tissue culture systems, our recent discovery of a murine norovirus (MNV) and its in vitro propagation have allowed us to begin addressing norovirus replication strategies and immune responses to norovirus infection. We have previously demonstrated that interferon responses are critical to control MNV-1 infection in vivo and to directly inhibit viral replication in vitro. We now extend these studies to define the molecular basis for interferon mediated inhibition. Viral replication intermediates were not detected in permissive cells pretreated with type I interferon after either infection or transfection of virion-associated RNA, demonstrating a very early block to virion production that is after virus entry and uncoating. A similar absence of viral replication intermediates was observed in infected primary macrophages and dendritic cells pretreated with type I IFN. This was not due to degradation of incoming genomes in interferon-pretreated cells since similar levels of genomes were present in untreated and pretreated cells through 6 h of infection, and these genomes retained their integrity. Surprisingly, this block to the translation of viral proteins was not dependent on the well-characterized interferon-induced antiviral molecule PKR. Similar results were observed in cells pretreated with type II interferon, except that the inhibition of viral translation was dependent on PKR. Thus, both type I and type II interferon signaling inhibit norovirus translation in permissive myeloid cells, but they display distinct dependence on PKR for this inhibition. PMID- 19297468 TI - Probing the transcription mechanisms of reovirus cores with molecules that alter RNA duplex stability. AB - The mammalian reovirus (MRV) genome comprises 10 double-stranded RNA (dsRNA) segments, packaged along with transcriptase complexes inside each core particle. Effects of four small molecules on transcription by MRV cores were studied for this report, chosen for their known capacities to alter RNA duplex stability. Spermidine and spermine, which enhance duplex stability, inhibited transcription, whereas dimethyl sulfoxide and trimethylglycine, which attenuate duplex stability, stimulated transcription. Different mechanisms were identified for inhibition or activation by these molecules. With spermidine, one round of transcription occurred normally, but subsequent rounds were inhibited. Thus, inhibition occurred at the transition between the end of elongation in one round and initiation in the next round of transcription. Dimethyl sulfoxide or trimethylglycine, on the other hand, had no effect on transcription by a constitutively active fraction of cores in each preparation but activated transcription in another fraction that was otherwise silent for the production of elongated transcripts. Activation of this other fraction occurred at the transition between transcript initiation and elongation, i.e., at promoter escape. These results suggest that the relative stability of RNA duplexes is most important for certain steps in the particle-associated transcription cycles of dsRNA viruses and that small molecules are useful tools for probing these and probably other steps. PMID- 19297467 TI - Restriction of human polyomavirus BK virus DNA replication in murine cells and extracts. AB - BK virus (BKV) causes persistent and asymptomatic infections in most humans and is the etiologic agent of polyomavirus-associated nephropathy (PVAN) and other pathologies. Unfortunately, there are no animal models with which to study activation of BKV replication in the human kidney and the accompanying PVAN. Here we report studies of the restriction of BKV replication in murine cells and extracts and the cause(s) of this restriction. Upon infection of murine cells, BKV expressed large T antigen (TAg), but viral DNA replication and progeny were not detected. Transfection of murine cells with BKV TAg expression vectors also caused TAg expression without accompanying DNA replication. Analysis of the replication of DNAs containing chimeric BKV and murine polyomavirus origins revealed the importance of BKV core origin sequences and TAg for DNA replication. A sensitive assay was developed with purified BKV TAg that supported TAg dependent BKV DNA replication with human but not with murine cell extracts. Addition of human replication proteins, DNA polymerase alpha-primase, replication protein A, or topoisomerase I to the murine extracts with BKV TAg did not rescue viral DNA replication. Notably, addition of murine extracts to human extracts inhibited BKV TAg-dependent DNA replication at a step prior to or during unwinding of the viral origin. These findings and differences in replication specificity between BKV TAg and the TAgs of simian virus 40 (SV40) and JC virus (JCV) and their respective origins implicate features of the BKV TAg and origin distinct from SV40 and JCV in restriction of BKV replication in murine cells. PMID- 19297469 TI - Residues in a highly conserved claudin-1 motif are required for hepatitis C virus entry and mediate the formation of cell-cell contacts. AB - Claudin-1, a component of tight junctions between liver hepatocytes, is a hepatitis C virus (HCV) late-stage entry cofactor. To investigate the structural and functional roles of various claudin-1 domains in HCV entry, we applied a mutagenesis strategy. Putative functional intracellular claudin-1 domains were not important. However, we identified seven novel residues in the first extracellular loop that are critical for entry of HCV isolates drawn from six different subtypes. Most of the critical residues belong to the highly conserved claudin motif W(30)-GLW(51)-C(54)-C(64). Alanine substitutions of these residues did not impair claudin-1 cell surface expression or lateral protein interactions within the plasma membrane, including claudin-1-claudin-1 and claudin-1-CD81 interactions. However, these mutants no longer localized to cell-cell contacts. Based on our observations, we propose that cell-cell contacts formed by claudin-1 may generate specialized membrane domains that are amenable to HCV entry. PMID- 19297470 TI - Helices alpha2 and alpha3 of West Nile virus capsid protein are dispensable for assembly of infectious virions. AB - The internal hydrophobic sequence within the flaviviral capsid protein (protein C) plays an important role in the assembly of infectious virions. Here, this sequence was analyzed in a West Nile virus lineage I isolate (crow V76/1). An infectious cDNA clone was constructed and used to introduce deletions into the internal hydrophobic domain which comprises helix alpha2 and part of the loop intervening helices alpha2 and alpha3. In total, nine capsid deletion mutants (4 to 14 amino acids long) were constructed and tested for virus viability. Some of the short deletions did not significantly affect growth in cell culture, whereas larger deletions removing almost the entire hydrophobic region significantly impaired viral growth. Efficient growth of the majority of mutants could, however, be restored by the acquisition of second-site mutations. In most cases, these resuscitating mutations were point mutations within protein C changing individual amino acids into more hydrophobic residues, reminiscent of what had been observed previously for another flavivirus, tick-borne encephalitis virus. However, we also identified viable spontaneous pseudorevertants with more than one-third of the capsid protein removed, i.e., 36 or 37 of a total of 105 residues, including all of helix alpha3 and a hydrophilic segment connecting alpha3 and alpha4. These large deletions are predicted to induce formation of large, predominantly hydrophobic fusion helices which may substitute for the loss of the internal hydrophobic domain, underlining the unrivaled structural and functional flexibility of protein C. PMID- 19297471 TI - Varicella-zoster virus infection induces autophagy in both cultured cells and human skin vesicles. AB - When grown in cultured cells, varicella-zoster virus (VZV) forms many aberrant light particles and produces low titers. Various studies have explored the reasons for such a phenotype and have pointed to impaired expression of specific late genes and at lysosomal targeting of egressing virions as possible causes. In the studies presented here, we report that the autophagic degradation pathway was induced at late time points after VZV infection of cultured cells, as documented by immunoblot analysis of the cellular proteins LC3B and p62/SQSTM1, along with electron microscopy analysis, which demonstrated the presence of both early autophagosomes and late autophagic compartments. Autophagy was induced in infected cells even in the presence of phosphonoacetic acid, an inhibitor of viral late gene expression, thus suggesting that accumulation of immediate-early and early viral gene products might be the major stimulus for its induction. We also showed that the autophagic response was not dependent on a specific cell substrate, virus strain, or type of inoculum. Finally, using immunofluorescence imaging, we demonstrated autophagosome-specific staining in human zoster vesicles but not in normal skin. Thus, our results document that this innate immune response pathway is a component of the VZV infectious cycle in both cultured cells and the human skin vesicle, the final site of virion formation in the infected human host. PMID- 19297472 TI - High-throughput cell-based screen for chemicals that inhibit infection by simian virus 40 and human polyomaviruses. AB - We developed a high-throughput, cell-based screen to identify chemicals that inhibit infection by the primate polyomaviruses. The screen is based on the detection of compounds that inhibit the ability of a replication-defective simian virus 40 (SV40)-based viral vector to cause growth arrest in HeLa cells by repressing the expression of the endogenous human papillomavirus E7 oncogene in these cells. We identified two compounds, ellagic acid and spiperone, that suppressed the ability of the SV40 recombinant virus to inhibit cellular DNA synthesis. These compounds caused a marked reduction of the ability of wild-type SV40 to productively infect permissive monkey cells, even when the compounds were added several hours after infection. The fraction of cells expressing SV40 large T antigen and the levels of T antigen mRNA were reduced in infected human and monkey cells treated with ellagic acid and spiperone, suggesting that these compounds block a step in the virus life cycle prior to SV40 early gene expression. Ellagic acid and spiperone also inhibited large T antigen expression by BK virus and JC virus, two important, pathogenic human polyomaviruses. PMID- 19297473 TI - Differential effect of human herpesvirus 6A on cell division and apoptosis among naive and central and effector memory CD4+ and CD8+ T-cell subsets. AB - The immune responses of naive and different memory subsets of CD4(+) and CD8(+) T cells to human herpesvirus 6 (HHV-6) have not been previously investigated. We show that HHV-6A induces cell division, as measured by 5,6-carboxyfluorescein succinimidyl ester dye and flow cytometry, predominantly in two populations of effector memory CD4(+) and CD8(+) T cells (T(EM) and T(EMRA)); naive (T(N)) and central memory (T(CM)) CD4(+) and CD8(+) T cells showed almost no cell division. In contrast, HHV-6A induced apoptosis primarily in T(N) and T(CM) CD4(+) and CD8(+) T cells, whereas T(EM) and T(EMRA) CD4(+) and CD8(+) T cells were resistant to HHV-6A-induced apoptosis. HHV-6A-induced apoptosis was associated with activation of caspase-8, caspase-9, and caspase-3, suggesting the involvement of death receptor and mitochondrial signaling pathways. In addition, HHV-6A induced secretion of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha), IL-8, and gamma interferon by peripheral blood mononuclear cells; TNF-alpha secretion was observed exclusively from CCR7(+) (T(N) plus T(CM)) CD4(+) T cells. These data show that HHV-6 differentially influences the functions of naive T cells and different subsets of memory CD4(+) and CD8(+) T cells, which in part may be due to differential susceptibility to HHV-6A-induced apoptosis. PMID- 19297474 TI - Induction of alpha/beta interferon by myxoma virus is selectively abrogated when primary mouse embryo fibroblasts become immortalized. AB - Mouse embryo fibroblasts (MEFs) are a widely used cell culture system in life sciences, including virology. Here, we show that although primary MEFs are nonpermissive to myxoma virus replication, the corresponding immortalized MEFs support a highly productive myxoma virus infection. We further demonstrate that this permissive phenotype for myxoma virus in immortalized MEFs is due to the immortalization-associated selective block to the cellular alpha/beta interferon induction machinery involved in responding to myxoma virus challenge. Thus, our report presents a clear example, illustrating that a drastic phenotypic alteration can occur with respect to virus infection between primary cells and their immortalized counterparts. PMID- 19297475 TI - Identification of integrin alpha3 as a new substrate of the adenovirus E4orf6/E1B 55-kilodalton E3 ubiquitin ligase complex. AB - The human adenovirus E4orf6 and E1B55K proteins promote viral replication by targeting several cellular proteins for degradation. The E4orf6 product has been shown by our group and others to form an E3 ubiquitin ligase complex that contains elongins B and C and cullin family member Cul5. E1B55K associates with this complex, where it is believed to function primarily to introduce bound substrates for degradation via proteasomes. In addition to p53, its first known substrate, the E4orf6/E1B 55-kDa complex (E4orf6/E1B55K) was shown to promote the degradation of Mre11 and DNA ligase IV; however, additional substrates are believed to exist. This notion is strengthened by the fact that none of these substrates seems likely to be associated with additional functions shown to be mediated by the E4orf6-associated E3 ubiquitin ligase complex, including export of late viral mRNAs and blockage of export of the bulk cellular mRNAs from the nucleus. In an attempt to identify new E4orf6/E1B55K substrates, we undertook a proteomic screen using human p53-null, non-small-cell lung carcinoma H1299 cells expressing either E4orf6 protein alone or in combination with E1B55K through infection by appropriate adenovirus vectors. One cellular protein that appeared to be degraded by E1B55K in combination with the E4orf6 protein was a species of molecular mass approximately 130 kDa that was identified as the integrin alpha3 subunit (i.e., very late activation antigen 3 alpha subunit). Preliminary analyses suggested that degradation of alpha3 may play a role in promoting release and spread of progeny virions. PMID- 19297476 TI - Cell-to-cell spread of the RNA interference response suppresses Semliki Forest virus (SFV) infection of mosquito cell cultures and cannot be antagonized by SFV. AB - In their vertebrate hosts, arboviruses such as Semliki Forest virus (SFV) (Togaviridae) generally counteract innate defenses and trigger cell death. In contrast, in mosquito cells, following an early phase of efficient virus production, a persistent infection with low levels of virus production is established. Whether arboviruses counteract RNA interference (RNAi), which provides an important antiviral defense system in mosquitoes, is an important question. Here we show that in Aedes albopictus-derived mosquito cells, SFV cannot prevent the establishment of an antiviral RNAi response or prevent the spread of protective antiviral double-stranded RNA/small interfering RNA (siRNA) from cell to cell, which can inhibit the replication of incoming virus. The expression of tombusvirus siRNA-binding protein p19 by SFV strongly enhanced virus spread between cultured cells rather than virus replication in initially infected cells. Our results indicate that the spread of the RNAi signal contributes to limiting virus dissemination. PMID- 19297477 TI - Recombinant Mycobacterium bovis BCG prime-recombinant adenovirus boost vaccination in rhesus monkeys elicits robust polyfunctional simian immunodeficiency virus-specific T-cell responses. AB - While mycobacteria have been proposed as vaccine vectors because of their persistence and safety, little has been done systematically to optimize their immunogenicity in nonhuman primates. We successfully generated recombinant Mycobacterium bovis BCG (rBCG) expressing simian immunodeficiency virus (SIV) Gag and Pol as multigenic, nonintegrating vectors, but rBCG-expressing SIV Env was unstable. A dose and route determination study in rhesus monkeys revealed that intramuscular administration of rBCG was associated with local reactogenicity, whereas intravenous and intradermal administration of 10(6) to 10(8) CFU of rBCG was well tolerated. After single or repeat rBCG inoculations, monkeys developed high-frequency gamma interferon enzyme-linked immunospot responses against BCG purified protein derivative. However, the same animals developed only modest SIV specific CD8(+) T-cell responses. Nevertheless, high-frequency SIV-specific cellular responses were observed in the rBCG-primed monkeys after boosting with recombinant adenovirus 5 (rAd5) expressing the SIV antigens. These cellular responses were of greater magnitude and more persistent than those generated after vaccination with rAd5 alone. The vaccine-elicited cellular responses were predominantly polyfunctional CD8(+) T cells. These findings support the further exploration of mycobacteria as priming vaccine vectors. PMID- 19297478 TI - Differential effects of hepatitis C virus JFH1 on human myeloid and plasmacytoid dendritic cells. AB - Dendritic cells (DCs) are reported to be functionally deficient during chronic hepatitis C virus (HCV) infection. Differing results have been reported on direct effects of intact replicative-form HCV on DC function. To better understand the effect of HCV on DC function, we treated freshly purified human myeloid DCs (mDCs) and plasmacytoid DCs (pDCs) with HCV JFH1. We found that HCV upregulated mDC maturation marker (CD83, CD86, and CD40) expression and did not inhibit Toll like receptor 3 (TLR3) ligand [poly(I:C)]-induced mDC maturation, a finding consistent with the phenotype of DCs from HCV-infected subjects. At the same time, HCV JFH1 inhibited the ability of poly(I:C)-treated mDCs to activate naive CD4 T cells. In contrast, although there was no direct effect of virus on pDC maturation, HCV JFH1 inhibited TLR7 ligand (R848)-induced pDC CD40 expression, and this was associated with impaired ability to activate naive CD4 T cells. Parallel experiments with recombinant HCV proteins indicated HCV core protein may be responsible for a portion of the activity. Furthermore, HCV-mediated mDC maturation was dependent upon CD81-E2 interaction and, in part, TLR2. Using UV treated HCV, we show that HCV-mediated mDC and pDC maturation is virus replication independent and, using strand specific PCR, we found no evidence for HCV replication within DCs. Because these effects of HCV on DC subset maturation and function in part recapitulate direct ex vivo analysis of DCs in chronic HCV infection, the mechanisms described here likely account for a portion of the DC subset defects observed in vivo. PMID- 19297479 TI - Differential virological and immunological outcome of severe acute respiratory syndrome coronavirus infection in susceptible and resistant transgenic mice expressing human angiotensin-converting enzyme 2. AB - We previously reported that transgenic (Tg) mice expressing human angiotensin converting enzyme 2 (hACE2), the receptor for severe acute respiratory syndrome coronavirus (SARS-CoV), were highly susceptible to SARS-CoV infection, which resulted in the development of disease of various severity and even death in some lineages. In this study, we further characterized and compared the pathogeneses of SARS-CoV infection in two of the most stable Tg lineages, AC70 and AC22, representing those susceptible and resistant to the lethal SARS-CoV infection, respectively. The kinetics of virus replication and the inflammatory responses within the lungs and brains, as well as the clinical and pathological outcomes, were assessed in each lineage. In addition, we generated information on lymphocyte subsets and mitogen-mediated proliferation of splenocytes. We found that while both lineages were permissive to SARS-CoV infection, causing elevated secretion of many inflammatory mediators within the lungs and brains, viral infection appeared to be more intense in AC70 than in AC22 mice, especially in the brain. Moreover, such infection was accompanied by a more profound immune suppression in the former, as evidenced by the extensive loss of T cells, compromised responses to concanavalin A stimulation, and absence of inflammatory infiltrates within the brain. We also found that CD8(+) T cells were partially effective in attenuating the pathogenesis of SARS-CoV infection in lethality resistant AC22 mice. Collectively, our data revealed a more intense viral infection and immunosuppression in AC70 mice than in AC22 mice, thereby providing us with an immunopathogenic basis for the fatal outcome of SARS-CoV infection in the AC70 mice. PMID- 19297480 TI - Identification of an intercistronic internal ribosome entry site in a Marek's disease virus immediate-early gene. AB - In this study, we have identified an internal ribosome entry site (IRES) from the highly infectious herpesvirus Marek's disease virus (MDV). The IRES was mapped to the intercistronic region (ICR) of a bicistronic mRNA that we cloned from the MDV transformed CD4(+) T-cell line MSB-1. The transcript is a member of a family of mRNAs expressed as immediate-early genes with two open reading frames (ORF). The first ORF encodes a 14-kDa polypeptide with two N-terminal splice variants, whereas the second ORF is contained entirely within a single exon and encodes a 12-kDa protein also known as RLORF9. We have shown that the ICR that separates the two ORFs functions as an IRES that controls the translation of RLORF9 when cap-dependent translation is inhibited. Deletion analysis revealed that there are two potential IRES elements within the ICR. Reverse genetic experiments with the oncogenic strain of MDV type 1 indicated that deletion of IRES-controlled RLORF9 does not significantly affect viral replication or MDV-induced mortality. PMID- 19297481 TI - CCR5- and CXCR4-tropic subtype C human immunodeficiency virus type 1 isolates have a lower level of pathogenic fitness than other dominant group M subtypes: implications for the epidemic. AB - Human immunodeficiency virus type 1 (HIV-1) subtype C is the dominant subtype globally, due largely to the incidence of subtype C infections in sub-Saharan Africa and east Asia. We compared the relative replicative fitness (ex vivo) of the major (M) group of HIV-1 subtypes A, B, C, D, and CRF01_AE and group O isolates. To estimate pathogenic fitness, pairwise competitions were performed between CCR5-tropic (R5) or CXCR4-tropic (X4) virus isolates in peripheral blood mononuclear cells (PBMC). A general fitness order was observed among 33 HIV-1 isolates; subtype B and D HIV-1 isolates were slightly more fit than the subtype A and dramatically more fit than the 12 subtype C isolates. All group M isolates were more fit (ex vivo) than the group O isolates. To estimate ex vivo transmission fitness, a subset of primary HIV-1 isolates were examined in primary human explants from penile, cervical, and rectal tissues. Only R5 isolates and no X4 HIV-1 isolates could replicate in these tissues, whereas the spread to PM1 cells was dependent on active replication and passive virus transfer. In tissue competition experiments, subtype C isolates could compete with and, in some cases, even win over subtype A and D isolates. However, when the migratory cells from infected tissues were mixed with a susceptible cell line, the subtype C isolates were outcompeted by other subtypes, as observed in experiments with PBMC. These findings suggest that subtype C HIV-1 isolates might have equal transmission fitness but reduced pathogenic fitness relative to other group M HIV 1 isolates. PMID- 19297482 TI - Acquisition of a polybasic hemagglutinin cleavage site by a low-pathogenic avian influenza virus is not sufficient for immediate transformation into a highly pathogenic strain. AB - Highly pathogenic avian influenza viruses (HPAIV) differ from all other strains by a polybasic cleavage site in their hemagglutinin. All these HPAIV share the H5 or H7 subtype. In order to investigate whether the acquisition of a polybasic cleavage site by an avirulent avian influenza virus strain with a hemagglutinin other than H5 or H7 is sufficient for immediate transformation into an HPAIV, we adapted the hemagglutinin cleavage site of A/Duck/Ukraine/1/1963 (H3N8) to that of the HPAIV A/Chicken/Italy/8/98 (H5N2), A/Chicken/HongKong/220/97 (H5N1), or A/Chicken/Germany/R28/03 (H7N7) and generated the recombinant wild-type and cleavage site mutants. In contrast to the wild type, multicycle replication of these mutants in tissue culture was demonstrated by positive plaque assays and viral multiplication in the absence of exogenous trypsin. Therefore, in vitro all cleavage site mutants resemble an HPAIV. However, in chicken they did not exhibit high pathogenicity, although they could be reisolated from cloacal swabs to some extent, indicating enhanced replication in vivo. These results demonstrate that beyond the polybasic hemagglutinin cleavage site, the virulence of HPAIV in chicken is based on additional pathogenicity determinants within the hemagglutinin itself or in the other viral proteins. Taken together, these observations support the notion that acquisition of a polybasic hemagglutinin cleavage site by an avirulent strain with a non-H5/H7 subtype is only one among several alterations necessary for evolution into an HPAIV. PMID- 19297483 TI - Herd immunity to GII.4 noroviruses is supported by outbreak patient sera. AB - Noroviruses (NoVs) of genogroup II, cluster 4 (GII.4), are the most common cause of outbreaks of acute gastroenteritis worldwide. During the past 13 years, GII.4 NoVs caused four seasons of widespread activity globally, each associated with the emergence of a new strain. In this report, we characterized the most recent epidemic strain, GII.4-2006 Minerva, by comparing virus-like particle (VLP) antigenic relationships and histo-blood group antigen (HBGA) binding profiles with strains isolated earlier. We also investigated the seroprevalence and specificity of GII.4 antibody in the years prior to, during, and following the GII.4 pandemic of 1995 and 1996 using a large collection of acute- and convalescent-phase serum pairs (n = 298) collected from 34 outbreaks. In a surrogate neutralization assay, we measured the blockade of HBGA binding using a panel of GII.4 VLPs representing strains isolated in 1987, 1997, 2002, and 2006 and a GII.3 VLP representing a strain isolated in the mid-1990s. Serum titers required for 50% HBGA blockade were compared between populations. In general, blockade of GII.4 VLP-HBGA binding was greater with convalescent-phase outbreak sera collected near the time of origin of the VLP strain. Heterotypic genotypes did not contribute to herd immunity against GII.4 NoVs based on their inability to block GII.4 VLP binding to HBGA. However, previous exposure to GII.4 NoV followed by infection by GII.3 NoV appeared to evoke an immune response to GII.4 NoV. These results support the hypothesis that herd immunity is a driving force for GII.4 evolution in the U.S. population. The data also suggest that complex patterns of cross-protection may exist across NoV genotypes in humans. PMID- 19297484 TI - The C terminus of the polerovirus p5 readthrough domain limits virus infection to the phloem. AB - Poleroviruses are restricted to vascular phloem tissues from which they are transmitted by their aphid vectors and are not transmissible mechanically. Phloem limitation has been attributed to the absence of virus proteins either facilitating movement or counteracting plant defense. The polerovirus capsid is composed of two forms of coat protein, the major P3 protein and the minor P3/P5 protein, a translational readthrough of P3. P3/P5 is required for insect transmission and acts in trans to facilitate long-distance virus movement in phloem tissue. Specific potato leafroll virus mutants lacking part or all of the P5 domain moved into and infected nonvascular mesophyll tissue when the source sink relationship of the plant (Solanum sarrachoides) was altered by pruning, with the progeny virus now being transmissible mechanically. However, in a period of months, a phloem-specific distribution of the virus was reestablished in the absence of aphid transmission. Virus from the new phloem-limited infection showed compensatory mutations that would be expected to restore the production of full length P3/P5 as well as the loss of mechanical transmissibility. The data support our hypothesis that phloem limitation in poleroviruses presumably does not result from a deficiency in the repertoire of virus genes but rather results from P3/P5 accumulation under selection in the infected plant, with the colateral effect of facilitating transmission by phloem-feeding aphid vectors. PMID- 19297485 TI - Prion protein glycosylation is not required for strain-specific neurotropism. AB - In this study, we tested the hypothesis that the glycosylation of the pathogenic isoform of the prion protein (PrP(Sc)) might encode the selective neurotropism of prion strains. We prepared unglycosylated cellular prion protein (PrP(C)) substrate molecules from normal mouse brain by treatment with PNGase F and used reconstituted serial protein cyclic misfolding amplification reactions to produce RML and 301C mouse prions containing unglycosylated PrP(Sc) molecules. Both RML- and 301C-derived prions containing unglycosylated PrP(Sc) molecules were infectious to wild-type mice, and neuropathological analysis showed that mice inoculated with these samples maintained strain-specific patterns of PrP(Sc) deposition and neuronal vacuolation. These results show that PrP(Sc) glycosylation is not necessary for strain-dependent prion neurotropism. PMID- 19297486 TI - A novel cardiotropic murine adenovirus representing a distinct species of mastadenoviruses. AB - During cell culture isolation experiments to recover Dobrava hantavirus from a suspension of liver from a striped field mouse (Apodemus agrarius), an unknown virus was coisolated. Atypically for hantaviruses, it had extensive cytopathic effects. Using a random PCR approach, it was identified as a novel murine adenovirus, MAdV-3 (for MAdV type 3). A plaque-purified virus clone was prepared and further characterized. The complete genome sequence of MAdV-3 was determined to be 30,570 bp in length. Sequence comparisons to other adenovirus species revealed highest similarity to MAdV-1, the representative of the murine adenovirus A species. However, substantial differences were found in the E1, E3, and E4 genomic regions. The phylogenetic distance of MAdV-3 amino acid sequences for pVIII, protease, polymerase, and hexon from MAdV-1 is markedly higher than 0.1 exchange per position, and, based on our cross-neutralization experiments, MAdV-3 and MAdV-1 can be regarded as different serotypes. Therefore, we propose to classify MAdV-3 as the first isolate of a novel adenovirus species, designated murine adenovirus C (MAdV-C). The novel MAdV-3 virus is not only genetically and serologically distinct from MAdV-1 but also shows a unique organ tropism in infected mice. In contrast to MAdV-1, the virus was not detectable in brain but predominantly infected heart tissue. Thus, infection of mice with cardiotropic MAdV-3 might be an interesting animal model of adenovirus-induced myocarditis. PMID- 19297487 TI - Suppression of bamboo mosaic virus accumulation by a putative methyltransferase in Nicotiana benthamiana. AB - Bamboo mosaic virus (BaMV) is a 6.4-kb positive-sense RNA virus belonging to the genus Potexvirus of the family Flexiviridae. The 155-kDa viral replicase, the product of ORF1, comprises an N-terminal S-adenosyl-l-methionine (AdoMet) dependent guanylyltransferase, a nucleoside triphosphatase/RNA 5'-triphosphatase, and a C-terminal RNA-dependent RNA polymerase (RdRp). To search for cellular factors potentially involved in the regulation of replication and/or transcription of BaMV, the viral RdRp domain was targeted as bait to screen against a leaf cDNA library of Nicotiana benthamiana using a yeast two-hybrid system. A putative methyltransferase (PNbMTS1) of 617 amino acid residues without an established physiological function was identified. Cotransfection of N. benthamiana protoplasts with a BaMV infectious clone and the PNbMTS1-expressing plasmid showed a PNbMTS1 dosage-dependent inhibitory effect on the accumulation of BaMV coat protein. Deletion of the N-terminal 36 amino acids, deletion of a predicted signal peptide or transmembrane segment, or mutations in the putative AdoMet-binding motifs of PNbMTS1 abolished the inhibitory effect. In contrast, suppression of PNbMTS1 by virus-induced gene silencing in N. benthamiana increased accumulation of the viral coat protein as well as the viral genomic RNA. Collectively, PNbMTS1 may function as an innate defense protein against the accumulation of BaMV through an uncharacterized mechanism. PMID- 19297488 TI - Characterization of a novel Golgi apparatus-localized latency determinant encoded by human cytomegalovirus. AB - Human cytomegalovirus (HCMV) exists indefinitely in infected individuals by a yet poorly characterized latent infection in hematopoietic cells. We previously demonstrated a requirement for the putative UL138 open reading frame (ORF) in promoting a latent infection in CD34(+) hematopoietic progenitor cells (HPCs) infected in vitro. In our present study, we have identified two coterminal transcripts of 2.7 and 3.6 kb and a 21-kilodalton (kDa) protein (pUL138) that are derived from the UL138 locus with early-late gene kinetics during productive infection. The UL138 transcripts and protein are detected in both fibroblasts and HPCs. A recombinant virus, FIX-UL138(STOP), that synthesizes the UL138 transcripts but not the protein exhibited a partial loss-of-latency phenotype in HPCs, similar to the phenotype observed for the UL138-null recombinant virus. This finding suggests that the UL138 protein is required for latency, but it does not exclude the possibility that the UL138 transcripts or other ORFs also contribute to latency. The mechanisms by which pUL138 contributes to latency remain unknown. While the 86- and 72-kDa immediate-early proteins were not detected in HPCs infected with HCMV in vitro, pUL138 did not function directly to suppress expression from the major immediate-early promoter in reporter assays. Interestingly, pUL138 localizes to the Golgi apparatus in infected cells but is not incorporated into virus particles. The localization of pUL138 to the Golgi apparatus suggests that pUL138 contributes to HCMV latency by a novel mechanism. pUL138 is the first HCMV protein demonstrated to promote an infection with the hallmarks of latency in CD34(+) HPCs. PMID- 19297489 TI - Recruitment of cdk9 to the immediate-early viral transcriptosomes during human cytomegalovirus infection requires efficient binding to cyclin T1, a threshold level of IE2 86, and active transcription. AB - Human cytomegalovirus (HCMV) infection results in the formation of nuclear viral transcriptosomes, which are sites dedicated to viral immediate-early (IE) transcription. At IE times of the infection, viral and cellular factors, including several components of transcription such as cyclin-dependent kinase 9 (cdk9), localize at these sites. To determine the mechanism and requirements of specific recruitment of cdk9 to the viral transcriptosomes, infection in the presence of inhibitor drugs and infection of cell lines expressing exogenous mutant cdk9 were performed. We found that cdk9 localization to the viral transcriptosomes requires de novo protein synthesis. In addition, active transcription is required for recruitment and maintenance of cdk9 at the viral transcriptosomes. In cells infected with a recombinant IE2 HCMV (IE2 86 DeltaSX virus) in which IE2 gene expression is greatly reduced, cdk9 localization at the transcriptosome is delayed and corresponds to the kinetics of accumulation of the IE2 protein at these sites. Infection in the presence of the cdk9 inhibitors Flavopiridol and DRB (5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole) allowed cdk9 localization to the viral transcriptosomes. A kinase-inactive cdk9 (D167N) expressed during the infection also localizes to the viral transcriptosomes, indicating that kinase activity of cdk9 is not a requirement for its localization to the sites of IE transcription. Exogenous expression of additional cdk9 mutants indicates that binding of Brd4 to the cdk9 complex is not required but that efficient binding to cyclin T1 is essential. PMID- 19297490 TI - Early control of H5N1 influenza virus replication by the type I interferon response in mice. AB - Widespread distribution of highly pathogenic avian H5N1 influenza viruses in domesticated and wild birds continues to pose a threat to public health, as interspecies transmission of virus has resulted in increasing numbers of human disease cases. Although the pathogenic mechanism(s) of H5N1 influenza viruses has not been fully elucidated, it has been suggested that the ability to evade host innate responses, such as the type I interferon response, may contribute to the virulence of these viruses in mammals. We investigated the role that type I interferons (alpha/beta interferon [IFN-alpha/beta]) might play in H5N1 pathogenicity in vivo, by comparing the kinetics and outcomes of H5N1 virus infection in IFN-alpha/beta receptor (IFN-alpha/betaR)-deficient and SvEv129 wild type mice using two avian influenza A viruses isolated from humans, A/Hong Kong/483/97 (HK/483) and A/Hong Kong/486/97 (HK/486), which exhibit high and low lethality in mice, respectively. IFN-alpha/betaR-deficient mice experienced significantly more weight loss and more rapid time to death than did wild-type mice. HK/486 virus caused a systemic infection similar to that with HK/483 virus in IFN-alpha/betaR-deficient mice, suggesting a role for IFN-alpha/beta in controlling the systemic spread of this H5N1 virus. HK/483 virus replicated more efficiently than HK/486 virus both in vivo and in vitro. However, replication of both viruses was significantly reduced following pretreatment with IFN alpha/beta. These results suggest a role for the IFN-alpha/beta response in the control of H5N1 virus replication both in vivo and in vitro, and as such it may provide some degree of protection to the host in the early stages of infection. PMID- 19297491 TI - Different evolutionary trajectories of European avian-like and classical swine H1N1 influenza A viruses. AB - In 1979, a lineage of avian-like H1N1 influenza A viruses emerged in European swine populations independently from the classical swine H1N1 virus lineage that had circulated in pigs since the Spanish influenza pandemic of 1918. To determine whether these two distinct lineages of swine-adapted A/H1N1 viruses evolved from avian-like A/H1N1 ancestors in similar ways, as might be expected given their common host species and origin, we compared patterns of nucleotide and amino acid change in whole genome sequences of both groups. An analysis of nucleotide compositional bias across all eight genomic segments for the two swine lineages showed a clear lineage-specific bias, although a segment-specific effect was also apparent. As such, there appears to be only a relatively weak host-specific selection pressure. Strikingly, despite each lineage evolving in the same species of host for decades, amino acid analysis revealed little evidence of either parallel or convergent changes. These findings suggest that although adaptation due to evolutionary lineages can be distinguished, there are functional and structural constraints on all gene segments and that the evolutionary trajectory of each lineage of swine A/H1N1 virus has a strong historical contingency. Thus, in the context of emergence of an influenza A virus strain via a host switch event, it is difficult to predict what specific polygenic changes are needed for mammalian adaptation. PMID- 19297492 TI - Early and strong immune responses are associated with control of viral replication and recovery in lassa virus-infected cynomolgus monkeys. AB - Lassa virus causes a hemorrhagic fever endemic in West Africa. The pathogenesis and the immune responses associated with the disease are poorly understood, and no vaccine is available. We followed virological, pathological, and immunological markers associated with fatal and nonfatal Lassa virus infection of cynomolgus monkeys. The clinical picture was characterized by fever, weight loss, depression, and acute respiratory syndrome. Transient thrombocytopenia and lymphopenia, lymphadenopathy, splenomegaly, infiltration of mononuclear cells, and alterations of the liver, lungs, and endothelia were observed. Survivors exhibited fewer lesions and a lower viral load than nonsurvivors. Although all animals developed strong humoral responses, antibodies appeared more rapidly in survivors and were directed against GP(1), GP(2), and NP. Type I interferons were detected early after infection in survivors but only during the terminal stages in fatalities. The mRNAs for CXCL10 (IP-10) and CXCL11 (I-TAC) were abundant in peripheral blood mononuclear cells and lymph nodes from infected animals, but plasma interleukin-6 was detected only in fatalities. In survivors, high activated-monocyte counts were followed by a rise in the total number of circulating monocytes. Activated T lymphocytes circulated in survivors, whereas T cell activation was low and delayed in fatalities. In vitro stimulation with inactivated Lassa virus induced activation of T lymphocytes from all infected monkeys, but only lymphocytes from survivors proliferated. Thus, early and strong immune responses and control of viral replication were associated with recovery, whereas fatal infection was characterized by major alterations of the blood formula and, in organs, weak immune responses and uncontrolled viral replication. PMID- 19297493 TI - Human immunodeficiency virus type 1 gp120 reprogramming of CD4+ T-cell migration provides a mechanism for lymphadenopathy. AB - Infection by human immunodeficiency virus type 1 (HIV-1) is associated with decreases in peripheral CD4(+) T cells and development of lymphadenopathy. The precise mechanisms by which HIV-1 induces these changes have not been elucidated. T-cell trafficking through lymphoid tissues is facilitated by CCL21-mediated entry and sphingosine-1-phosphate (S1P)-mediated egress. Having previously determined that HIV-1 envelop glycoprotein, gp120, directly alters T-cell migration, we investigated whether gp120 without HIV-1 infection could influence the responses of CD4(+) T cells to the signals involved in T-cell trafficking through lymph tissue. Incubation of normal human T cells with gp120 for 1 h resulted in reprogramming of CD4 T-cell migratory responses by increasing sensitivity to CCL20 and CCL21 and complete inhibition of migration to S1P. Incubation of human T cells with gp120 prior to injection into NOD.CB17 Prkdc(scid)/J mice resulted in increases in lymph node accumulation of CD4(+) T cells, with reciprocal decreases in blood and spleen compared to T cells not exposed to gp120. The effects of gp120 required CD4 signaling mediated through p56(lck). These findings suggest that gp120 alone can alter CD4(+) influx and efflux from lymph nodes in a fashion consistent with the development of lymphopenia and lymphadenopathy. PMID- 19297494 TI - Regulation of the catalytic activity of herpes simplex virus 1 protein kinase Us3 by autophosphorylation and its role in pathogenesis. AB - Us3 is a serine/threonine protein kinase encoded by herpes simplex virus 1 (HSV 1). We recently identified serine at Us3 position 147 (Ser-147) as a physiological phosphorylation site of Us3 (A. Kato, M. Tanaka, M. Yamamoto, R. Asai, T. Sata, Y. Nishiyama, and Y. Kawaguchi, J. Virol. 82:6172-6189, 2008). In the present study, we investigated the effects of phosphorylation of Us3 Ser-147 on regulation of Us3 catalytic activity in infected cells and on HSV-1 pathogenesis. Our results were as follows. (i) Only a small fraction of Us3 purified from infected cells was phosphorylated at Ser-147. (ii) Us3 phosphorylated at Ser-147 purified from infected cells had significantly higher kinase activity than Us3 not phosphorylated at Ser-147. (iii) Phosphorylation of Us3 Ser-147 in infected cells was dependent on Us3 kinase activity. (iv) Replacement of Us3 Ser-147 by alanine significantly reduced viral replication in the mouse cornea and the development of herpes stromal keratitis and periocular skin disease in mice. These results indicated that Us3 catalytic activity is tightly regulated by autophosphorylation of Ser-147 in infected cells and that regulation of Us3 activity by autophosphorylation appeared to play a critical role in viral replication in vivo and in HSV-1 pathogenesis. PMID- 19297495 TI - Aquareovirus effects syncytiogenesis by using a novel member of the FAST protein family translated from a noncanonical translation start site. AB - As nonenveloped viruses, the aquareoviruses and orthoreoviruses are unusual in their ability to induce cell-cell fusion and syncytium formation. While an extraordinary family of fusion-associated small transmembrane (FAST) proteins is responsible for orthoreovirus syncytiogenesis, the basis for aquareovirus-induced syncytiogenesis is unknown. We now report that the S7 genome segment of an Atlantic salmon reovirus is polycistronic and uses a noncanonical CUG translation start codon to produce a 22-kDa integral membrane protein responsible for syncytiogenesis. The aquareovirus p22 protein represents a fourth distinct member of the FAST family with a unique repertoire and arrangement of structural motifs. PMID- 19297496 TI - Broad tissue and cell tropism of avian bornavirus in parrots with proventricular dilatation disease. AB - Avian bornaviruses (ABV), representing a new genus within the family Bornaviridae, were recently discovered in parrots from North America and Israel with proventricular dilatation disease (PDD). We show here that closely related viruses are also present in captive European parrots of various species with PDD. The six ABV strains that we identified in clinically diseased birds are new members of the previously defined ABV genotypes 2 and 4. Viruses of both genotypes readily established persistent, noncytolytic infections in quail and chicken cell lines but did not grow in cultured mammalian cells in which classical Borna disease virus strains replicate very efficiently. ABV antigens were present in both the cytoplasm and nucleus of infected cells, suggesting nuclear replication of ABV. The genome organization of avian and mammalian bornaviruses is highly conserved except that ABV lacks a distinct control element in the 5' noncoding region of the bicistronic mRNA encoding the viral proteins X and P. Reverse transcription-PCR analysis demonstrated the presence of virus in many, if not all, organs of birds with PDD. Viral nucleic acid was also found in feces of diseased birds, suggesting virus transmission by the fecal-oronasal route. Immunohistochemical analysis of organs from birds with PDD revealed that infection with ABV is not restricted to cells of the nervous system. Thus, ABV exhibits a broad tissue and cell tropism that is strikingly different from classical Borna disease virus. PMID- 19297497 TI - DDX3 DEAD-Box RNA helicase inhibits hepatitis B virus reverse transcription by incorporation into nucleocapsids. AB - Viruses utilize host factors in many steps of their life cycles. Yet, little is known about host factors that contribute to the life cycle of hepatitis B virus (HBV), which replicates its genome by reverse transcription. To identify host factors that contribute to viral reverse transcription, we sought to identify cellular proteins that interact with HBV polymerase (Pol) by using affinity purification coupled with mass spectrometry. One of the HBV Pol-interacting host factors identified was DDX3 DEAD-box RNA helicase, which unwinds RNA in an ATPase dependent manner. Recently, it was shown that DDX3 is essential for both human immunodeficiency virus and hepatitis C virus infection. In contrast, we found that the ectopic expression of DDX3 led to significantly reduced viral DNA synthesis. The DDX3-mediated inhibition of viral DNA synthesis did not affect RNA encapsidation, a step prior to reverse transcription, and indicated that DDX3 inhibits HBV reverse transcription. Mutational analysis revealed that mutant DDX3 with an inactive ATPase motif, but not that with an inactive RNA helicase motif, failed to inhibit viral DNA synthesis. Our interpretation is that DDX3 inhibits viral DNA synthesis at a step following ATP hydrolysis but prior to RNA unwinding. Finally, OptiPrep density gradient analysis revealed that DDX3 was incorporated into nucleocapsids, suggesting that DDX3 inhibits viral reverse transcription following nucleocapsid assembly. Thus, DDX3 represents a novel host restriction factor that limits HBV infection. PMID- 19297498 TI - Impaired cholesterol biosynthesis in a neuronal cell line persistently infected with measles virus. AB - Measles virus remains a substantial cause of morbidity and mortality, producing acute infection with a potential for development of viral persistence. To study the events underlying acute and persistent measles virus infection, we performed a global transcriptional analysis on murine neuroblastoma cells that were acutely or persistently infected with measles virus. In general, we found that acute infection induced significantly more gene expression changes than did persistent infection. A functional enrichment analysis to identify which host pathways were perturbed during each of these infections identified several pathways related to cholesterol biosynthesis, including cholesterol metabolic processes, hydroxymethylglutaryl-coenzyme A (CoA) reductase activity, and acetyl-CoA C acetyltransferase activity. We also found that measles virus colocalized to lipid rafts in both acute and persistent infection models and that the majority of genes associated with cholesterol synthesis were downregulated in persistent infection relative to acute infection, suggesting a possible link with the defective viral budding in persistent infection. Further, we found that pharmacological inhibition of cholesterol synthesis resulted in the inhibition of viral budding during acute infection. In summary, persistent measles viral infection was associated with decreased cholesterol synthesis, a lower abundance of cholesterol and lipid rafts in the cell membrane, and inhibition of giant-cell formation and release of viral progeny. PMID- 19297499 TI - Evidence that productive human immunodeficiency virus type 1 assembly can occur in an intracellular compartment. AB - Human immunodeficiency virus type 1 (HIV-1) assembly occurs predominantly at the plasma membrane of infected cells. The targeting of assembly to intracellular compartments such as multivesicular bodies (MVBs) generally leads to a significant reduction in virus release efficiency, suggesting that MVBs are a nonproductive site for HIV-1 assembly. In the current study, we make use of an HIV-1 Gag-matrix mutant, 29/31KE, that is MVB targeted. We previously showed that this mutant is severely defective for virus particle production in HeLa cells but more modestly affected in primary macrophages. To more broadly examine the consequences of MVB targeting for virus production, we investigated 29/31KE particle production in a range of cell types. Surprisingly, this mutant supported highly efficient assembly and release in T cells despite its striking MVB Gag localization. Manipulation of cellular endocytic pathways revealed that unlike Vpu-defective HIV-1, which demonstrated intracellular Gag localization as a result of Gag endocytosis from the plasma membrane, 29/31KE mutant Gag was targeted directly to an MVB compartment. The 29/31KE mutant was unable to support multiple-round replication; however, this defect could be reversed by truncating the cytoplasmic tail of the transmembrane envelope glycoprotein gp41 and by the acquisition of a 16EK change in matrix. The 16EK/29/31KE matrix mutant replicated efficiently in the MT-4 T-cell line despite maintaining an MVB-targeting phenotype. These results indicate that MVB-targeted Gag can be efficiently released from T cells and primary macrophages, suggesting that under some circumstances, late endosomal compartments can serve as productive sites for HIV 1 assembly in these physiologically relevant cell types. PMID- 19297500 TI - Biochemical characterization of arterivirus nonstructural protein 11 reveals the nidovirus-wide conservation of a replicative endoribonuclease. AB - Nidoviruses (arteriviruses, coronaviruses, and roniviruses) are a phylogenetically compact but diverse group of positive-strand RNA viruses that includes important human and animal pathogens. Nidovirus RNA synthesis is mediated by a cytoplasmic membrane-associated replication/transcription complex that includes up to 16 viral nonstructural proteins (nsps), which carry common enzymatic activities, like the viral RNA polymerase, but also unusual and poorly understood RNA-processing functions. Of these, a conserved endoribonuclease (NendoU) is a major genetic marker that is unique to nidoviruses. NendoU activity was previously verified in vitro for the coronavirus nsp15, but not for any of its distantly related orthologs from other nidovirus lineages, like the arterivirus nsp11. Here, we show that the bacterially expressed nsp11 proteins of two arteriviruses, equine arteritis virus and porcine respiratory and reproductive syndrome virus, possess pyrimidine-specific endoribonuclease activity. RNA cleavage was independent of divalent cations in vitro and was greatly reduced by replacement of residues previously implicated in catalysis. Comparative characterization of the NendoU activity in arteriviruses and severe acute respiratory syndrome coronavirus revealed common and distinct features of their substrate requirements and reaction mechanism. Our data provide the first biochemical evidence of endoribonuclease activity associated with arterivirus nsp11 and support the conclusion that this remarkable RNA-processing enzyme, whose substrate in the infected cell remains to be identified, distinguishes nidoviruses from all other RNA viruses. PMID- 19297501 TI - Optimal translation initiation enables Vif-deficient human immunodeficiency virus type 1 to escape restriction by APOBEC3G. AB - APOBEC3G restricts Vif-deficient human immunodeficiency virus type 1 (HIV-1) by deaminating viral cDNA cytosines to uracils. This promutagenic activity is counteracted by HIV-1 Vif, which is a natural APOBEC3G antagonist. However, we previously reported that Vif-deficient HIV-1 could evolve resistance to APOBEC3G by a novel mechanism requiring an A200-to-C/T transition mutation and Vpr inactivation. A pyrimidine at nucleotide 200 in the untranslated leader region contributed to resistance by increasing virus particle production, which resulted in fewer APOBEC3G molecules per particle. Here we show that the A200-to-C/T mutation functions posttranscriptionally by inactivating an upstream start codon, which in turn enables optimal viral mRNA translation from canonical start codons. PMID- 19297502 TI - Effect of transplantation of bone marrow-derived mesenchymal stem cells on mice infected with prions. AB - Bone marrow-derived mesenchymal stem cells (MSCs) have been reported to migrate to brain lesions in experimental models of ischemia, tumors, and neurodegenerative diseases and to ameliorate functional deficits. In this study, we attempted to evaluate the therapeutic potential of MSCs for treating prion diseases. Immortalized human MSCs (hMSCs) that express the LacZ gene were transplanted into the unilateral hippocampi or thalami of mice, and their distributions were monitored by the expression of beta-galactosidase. In mice infected with prions, hMSCs transplanted at 120 days postinoculation (dpi) were detected on the contralateral side at 2 days after transplantation and existed there even at 3 weeks after transplantation. In contrast, few hMSCs were detected on the contralateral side for mock-infected mice. Interestingly, the migration of hMSCs appeared to correlate with the severity of neuropathological lesions, including disease-specific prion protein deposition. The hMSCs also migrated to a prion-specific lesion in the brain, even when intravenously injected. Although the effects were modest, intrahippocampal and intravenous transplantation of hMSCs prolonged the survival of mice infected with prions. A subpopulation of hMSCs in the brains of prion-infected mice produced various trophic factors and differentiated into cells of neuronal and glial lineages. These results suggest that MSCs have promise as a cellular vehicle for the delivery of therapeutic genes to brain lesions associated with prion diseases and, furthermore, that they may help to regenerate neuronal tissues damaged by prion propagation. PMID- 19297503 TI - Polyfunctional CD4+ T-cell induction in neutralizing antibody-triggered control of simian immunodeficiency virus infection. AB - Rapid depletion of memory CD4(+) T cells and delayed induction of neutralizing antibody (NAb) responses are characteristics of human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infections. Although it was speculated that postinfection NAb induction could have only a limited suppressive effect on primary HIV replication, a recent study has shown that a single passive NAb immunization of rhesus macaques 1 week after SIV challenge can result in reduction of viral loads at the set point, indicating a possible contribution of postinfection NAb responses to virus control. However, the mechanism accounting for this NAb-triggered SIV control has remained unclear. Here, we report rapid induction of virus-specific polyfunctional T-cell responses after the passive NAb immunization postinfection. Analysis of SIV Gag-specific responses of gamma interferon, tumor necrosis factor alpha, interleukin-2, macrophage inflammatory protein 1beta, and CD107a revealed that the polyfunctionality of Gag-specific CD4(+) T cells, as defined by the multiplicity of these responses, was markedly elevated in the acute phase in NAb-immunized animals. In the chronic phase, despite the absence of detectable NAbs, virus control was maintained, accompanied by polyfunctional Gag-specific T-cell responses. These results implicate virus specific polyfunctional CD4(+) T-cell responses in this NAb-triggered virus control, suggesting possible synergism between NAbs and T cells for control of HIV/SIV replication. PMID- 19297504 TI - Virology in the 21st century. PMID- 19297505 TI - Insular lesionectomy for refractory epilepsy: management and outcome. AB - Surgical treatment of deep-seated insular lesions causing refractory epilepsy is thought to be difficult due to the complicated accessibility and close proximity of eloquent areas. Here we report our experience with insular lesionectomies. Twenty-four patients (range 1-62 years, mean 27) who underwent epilepsy-surgery for a lesion involving the insular region, were identified from the epilepsy surgery data bank. We analysed pre-surgical diagnostics, surgical strategy and postoperative follow up concerning functional morbidity and seizure outcome (range 12-168 months, mean 37.5). Eight patients had pure insular lesions, in 16 cases the lesion extended either to the frontal (n = 3) or temporal lobe (n = 8) or was multilobar (n = 5). Sixteen resections (66.7%) were done on the right side. Six patients required invasive EEG-recording, three patients received intra operative electrocorticography. In seven patients only subtotal resection of the insular lesion was possible due to involvement of eloquent areas. Thirteen patients suffered from glial/glioneural tumours (WHO grades I-III), 11 from non neoplastic lesions. Postoperatively, one patient had a hemihypesthesia and one patient had a deterioration of a pre-existing hemiparesis; two patients had a hemianopia as calculated deficit (mild permanent morbidity 16.6%). According to the ILAE-classification, 15 patients were completely seizure free (62.5%, ILAE 1). Around 79.2% had satisfactory seizure outcome (ILAE 1-3). In selected patients an individually tailored lesionectomy of insular lesions can be performed, which is acceptably safe and provides a high rate of satisfactory seizure relief. Even subtotal resection can result in good seizure control. PMID- 19297506 TI - The clinical profile of right temporal lobe atrophy. AB - Frontotemporal lobar degeneration is currently associated with three syndromic variants. Disorders of speech and language figure prominently in two of the three variants, and are associated with left-sided frontotemporal atrophy. The detailed characterization of these syndromes contrasts with the relative paucity of information relating to frontotemporal lobar degeneration primarily affecting the right cerebral hemisphere. The objective of this study was to identify the clinical profile associated with asymmetrical, predominantly right-sided, temporal lobe atrophy. Twenty patients with predominant right temporal lobe atrophy were identified on the basis of blinded visual assessment of the MRI scans. The severity of right temporal lobe atrophy was quantified using volumetric analysis of the whole temporal lobes, the amygdala and the hippocampus. Profiles of cognitive function, behavioural and personality changes were obtained on each patient. The pattern of atrophy and the clinical features were compared with those observed in a group of patients with semantic dementia and predominant left-sided temporal lobe atrophy. The mean right temporal lobe volume in the right temporal lobe atrophy group was reduced by 37%, with the mean left temporal lobe volume reduced by 19%. There was marked atrophy of the right hippocampus and right amygdala, with mean volumes reduced by 41 and 51%, respectively (left hippocampus and amygdala volumes were reduced by 18 and 33%, respectively). The most prominent cognitive deficits were impairment of episodic memory and getting lost. Prosopagnosia was a symptom in right temporal lobe atrophy patients. These patients also exhibited a variety of behavioural symptoms including social disinhibition, depression and aggressive behaviour. Nearly all behavioural disorders were more prevalent in the right temporal lobe atrophy patient group than the semantic dementia group. Symptoms particular to the right temporal lobe atrophy patient group included hyper-religiosity, visual hallucinations and cross-modal sensory experiences. The combination of clinical features associated with predominant right temporal lobe atrophy differs significantly from those associated with the other syndromes associated with focal degeneration of the frontal and temporal lobes and it is, therefore, proposed that this right temporal variant should be considered a separate syndromic variant of frontotemporal lobar degeneration. PMID- 19297507 TI - High frequency oscillations in intracranial EEGs mark epileptogenicity rather than lesion type. AB - High frequency oscillations (HFOs) called ripples (80-250 Hz) and fast ripples (FR, 250-500 Hz) can be recorded from intracerebral EEG macroelectrodes in patients with intractable epilepsy. HFOs occur predominantly in the seizure onset zone (SOZ) but their relationship to the underlying pathology is unknown. It was the aim of this study to investigate whether HFOs are specific to the SOZ or result from pathologically changed tissue, whether or not it is epileptogenic. Patients with different lesion types, namely mesial temporal atrophy (MTA), focal cortical dysplasia (FCD) and nodular heterotopias (NH) were investigated. Intracranial EEG was recorded from depth macroelectrodes with a sampling rate of 2000 Hz. Ripples (80-250 Hz) and Fast Ripples (250-500 Hz) were visually marked in 12 patients: five with MTA, four with FCD and three with NH. Rates of events were statistically compared in channels in four areas: lesional SOZ, non-lesional SOZ, lesional non-SOZ and non-lesional non-SOZ. HFO rates were clearly more linked to the SOZ than to the lesion. They were highest in areas in which lesion and SOZ overlap, but in patients with a SOZ outside the lesion, such as in NHs, HFO rates were clearly higher in the non-lesional SOZ than in the inactive lesions. No specific HFO pattern could be identified for the different lesion types. The findings suggest that HFOs represent a marker for SOZ areas independent of the underlying pathology and that pathologic tissue changes alone do not lead to high rates of HFOs. PMID- 19297508 TI - Sensorimotor dysfunction in multiple sclerosis and column-specific magnetization transfer-imaging abnormalities in the spinal cord. AB - The human spinal cord contains segregated sensory and motor pathways that have been difficult to quantify using conventional magnetic resonance imaging (MRI) techniques. Multiple sclerosis is characterized by both focal and spatially diffuse spinal cord lesions with heterogeneous pathologies that have limited attempts at linking MRI and behaviour. We used a novel magnetization-transfer weighted imaging approach to quantify damage to spinal white matter columns and tested its association with sensorimotor impairment. We studied 42 participants with multiple sclerosis who each underwent MRI at 3 Tesla and quantitative tests of sensorimotor function. We measured cerebrospinal-fluid-normalized magnetization-transfer signals in the dorsal and lateral columns and grey matter of the cervical cord. We also measured brain lesion volume, cervical spinal cord lesion number and cross-sectional area, vibration sensation, strength, walking velocity and standing balance. We used linear regression to assess the relationship between sensorimotor impairment and MRI abnormalities. We found that the dorsal column cerebrospinal-fluid-normalized magnetization-transfer signal specifically correlated with vibration sensation (R = 0.58, P < 0.001) and the lateral column signal with strength (R = -0.45, P = 0.003). Spinal cord signal measures also correlated with walking and balance dysfunction. A stepwise multiple regression showed that the dorsal column signal and diagnosis subtype alone explained a significant portion of the variance in sensation (R(2) = 0.54, P < 0.001), whereas the lateral column signal and diagnosis subtype explained a significant portion of the variance in strength (R(2) = 0.30, P < 0.001). These results help to understand the anatomic basis of sensorimotor disability in multiple sclerosis and have implications for testing the effects of neuroprotective and reparative interventions. PMID- 19297509 TI - Pontine-ventral respiratory column interactions through raphe circuits detected using multi-array spike train recordings. AB - Recently, Segers et al. identified functional connectivity between the ventrolateral respiratory column (VRC) and the pontine respiratory group (PRG). The apparent sparseness of detected paucisynaptic interactions motivated consideration of other potential functional pathways between these two regions. We report here evidence for "indirect" serial functional linkages between the PRG and VRC via intermediary brain stem midline raphe neurons. Arrays of microelectrodes were used to record sets of spike trains from a total of 145 PRG, 282 VRC, and 340 midline neurons in 11 decerebrate, vagotomized, neuromuscularly blocked, ventilated cats. Spike trains of 13,843 pairs of neurons that included at least one raphe cell were screened for respiratory modulation and short-time scale correlations. Significant correlogram features were detected in 7.2% of raphe-raphe (291/4,021), 4.3% of VRC-raphe (292/6,755), and 4.0% of the PRG-raphe (124/3,067) neuron pairs. Central peaks indicative of shared influences were the most common feature in correlations between pairs of raphe neurons, whereas correlated raphe-PRG and raphe-VRC neuron pairs displayed predominantly offset peaks and troughs, features suggesting a paucisynaptic influence of one neuron on the other. Overall, offset correlogram features provided evidence for 33 VRC-to raphe-to-PRG and 45 PRG-to-raphe-to-VRC correlational linkage chains with one or two intermediate raphe neurons. The results support a respiratory network architecture with parallel VRC-to-PRG and PRG-to-VRC links operating through intervening midline circuits, and suggest that raphe neurons contribute to the respiratory modulation of PRG neurons and shape the respiratory motor pattern through coordinated divergent actions on both the PRG and VRC. PMID- 19297510 TI - Functional interaction between TRPV1 and mu-opioid receptors in the descending antinociceptive pathway activates glutamate transmission and induces analgesia. AB - The transient receptor potential vanilloid-1 (TRPV1) receptor is involved in peripheral and spinal nociceptive processing and is a therapeutic target for pain. We have shown previously that TRPV1 in the ventrolateral periaqueductal gray (VL-PAG) tonically contributes to brain stem descending antinociception by stimulating glutamate release into the rostral ventromedial medulla and off neuron activity. Because both opioid and vanilloid systems integrate and transduce pain sensation in these pathways, we studied the potential interaction between TRPV1 and mu-opioid receptors in the VL-PAG-rostral ventromedial medulla (RVM) system. We found that the TRPV1 agonist, capsaicin, and the mu-receptor agonist [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]enkephalin, when coadministered into the ventrolateral-PAG at doses nonanalgesic per se, produce 1) antinociception in tests of thermal nociception; 2) stimulation of glutamate release into the RVM; and 3) inhibition of on neuron activity in the RVM. These effects were all antagonized by the TRPV1 and opioid receptor antagonists 5'-iodo-resiniferatoxin and naloxone, respectively, thus suggesting the existence of a TRPV1-mu-opioid interaction in the VL-PAG-RVM system. By using double immunofluorescence techniques, we found that TRPV1 and mu-opioid receptors are coexpressed in several neurons of the VL-PAG. These findings suggest that mu-receptor activation not only acts on inhibitory neurons to disinhibit PAG output neurons but also interacts with TRPV1 activation at increasing glutamate release into the RVM, possibly by acting directly on PAG output neurons projecting to the RVM. PMID- 19297511 TI - Modulation of spontaneous and odorant-evoked activity of rat olfactory sensory neurons by two anorectic peptides, insulin and leptin. AB - In mammals, the sense of smell is modulated by the status of satiety, which is mainly signaled by blood-circulating peptide hormones. However, the underlying mechanisms linking olfaction and food intake are poorly understood. Here we investigated the effects of two anorectic peptides, insulin and leptin, on the functional properties of olfactory sensory neurons (OSNs). Using patch-clamp recordings, we analyzed the spontaneous activity of rat OSNs in an in vitro intact epithelium preparation. Bath perfusion of insulin and leptin significantly increased the spontaneous firing frequency in 91.7% (n = 24) and 75.0% (n = 24) of the cells, respectively. When the activity was electrically evoked, both peptides shortened the latency to the first action potential by approximately 25% and decreased the interspike intervals by approximately 13%. While insulin and leptin enhanced the electrical excitability of OSNs in the absence of odorants, they surprisingly reduced the odorant-induced activity in the olfactory epithelium. Insulin and leptin decreased the peak amplitudes of isoamyl acetate induced electroolfactogram (EOG) signals to 46 and 38%, respectively. When measured in individual cells by patch-clamp recordings, insulin and leptin decreased odorant-induced transduction currents and receptor potentials. Therefore by increasing the spontaneous activity but reducing the odorant-induced activity of OSNs, an elevated insulin and leptin level (such as after a meal) may result in a decreased global signal-to-noise ratio in the olfactory epithelium, which matches the smell ability to the satiety status. PMID- 19297512 TI - Dynamics of primate oculomotor plant revealed by effects of abducens microstimulation. AB - Despite their importance for deciphering oculomotor commands, the mechanics of the extraocular muscles and orbital tissues (oculomotor plant) are poorly understood. In particular, the significance of plant nonlinearities is uncertain. Here primate plant dynamics were investigated by measuring the eye movements produced by stimulating the abducens nucleus with brief pulse trains of varying frequency. Statistical analysis of these movements indicated that the effects of stimulation lasted about 40 ms after the final pulse, after which the eye returned passively toward its position before stimulation. Behavior during the passive phase could be approximated by a linear plant model, corresponding to Voigt elements in series, with properties independent of initial eye position. In contrast, behavior during the stimulation phase revealed a sigmoidal relation between stimulation frequency and estimated steady-state tetanic tension, together with a frequency-dependent rate of tension increase, that appeared very similar to the nonlinearities previously found for isometric-force production in primate lateral rectus muscle. These results suggest that the dynamics of the oculomotor plant have an approximately linear component related to steady-state viscoelasticity and a nonlinear component related to changes in muscle activation. The latter may in part account for the nonlinear relations observed between eye-movement parameters and single-unit firing patterns in the abducens nucleus. These findings point to the importance of recruitment as a simplifying factor for motor control with nonlinear plants. PMID- 19297513 TI - Memory retention and spike-timing-dependent plasticity. AB - Memory systems should be plastic to allow for learning; however, they should also retain earlier memories. Here we explore how synaptic weights and memories are retained in models of single neurons and networks equipped with spike-timing dependent plasticity. We show that for single neuron models, the precise learning rule has a strong effect on the memory retention time. In particular, a soft bound, weight-dependent learning rule has a very short retention time as compared with a learning rule that is independent of the synaptic weights. Next, we explore how the retention time is reflected in receptive field stability in networks. As in the single neuron case, the weight-dependent learning rule yields less stable receptive fields than a weight-independent rule. However, receptive fields stabilize in the presence of sufficient lateral inhibition, demonstrating that plasticity in networks can be regulated by inhibition and suggesting a novel role for inhibition in neural circuits. PMID- 19297514 TI - Polyneuronal innervation of single muscle fibers in cat eye muscle: inferior oblique. AB - Single muscle fibers with multiple axonal endplates (multiply innervated fibers) are normally present in adult extraocular muscles (EOMs), while most other mammalian skeletal muscles contain fibers with a single myoneural junction. Recent findings by others led us to investigate for the presence of polyneuronal innervation (innervation of a single muscle fiber by >1 motoneuron) in the inferior oblique (IO) muscle of pentobarbital anesthetized cats. The IO muscle nerve branches, as they coursed through the orbit, were further divided for independent or simultaneous electrical stimulation with bipolar electrodes. Four of five established tests for polyneuronal innervation gave positive results. The sum of the twitch (1) and tetanic (2) tensions in response to individual nerve branch stimulation was greater than that for simultaneous (whole) nerve stimulation. The summed electromyographic (EMG) responses (3) gave a similar positive result. The result for crossed tetanic potentiation (4) was negative for polyneuronal innervation while the crossed fatigue (5) test was positive. These results are consistent with recent studies. That the EOMs exhibit polyneuronal innervation further explains the eye-movement system's functional integrity during some neuromuscular disorders as well as its ability to operate with precision after the loss of numerous motoneurons. PMID- 19297515 TI - Bumetanide, an NKCC1 antagonist, does not prevent formation of epileptogenic focus but blocks epileptic focus seizures in immature rat hippocampus. AB - Excitatory GABA action induced by high [Cl(-)](i) is thought to contribute to seizure generation in neonatal neurons although the mechanism of this effect remains unclear. We report that bumetanide, a NKCC1 antagonist, reduces driving force of GABA-mediated currents (DF(GABA)) in neonatal hippocampal neurons and blocks the giant depolarizing potentials (GDPs), a spontaneous pattern of network activity. In the preparation composed of two intact interconnected hippocampi, bumetanide did not prevent generation of kainate-induced seizures, their propagation to the contralateral hippocampus, and formation of an epileptogenic mirror focus. However, in the isolated mirror focus, bumetanide effectively blocked spontaneous epileptiform activity transforming it to the GDP-like activity pattern. Bumetanide partially reduced DF(GABA) and therefore the excitatory action of GABA in epileptic neurons. Therefore bumetanide is a potent anticonvulsive agent although it cannot prevent formation of the epileptogenic mirror focus. We suggest that an additional mechanism other than NKCC1-mediated contributes to the persistent increase of DF(GABA) in epileptic neurons. PMID- 19297516 TI - Cocaine-induced LTP in the ventral tegmental area: new insights into mechanism and time course illuminate the cellular substrates of addiction. AB - Previous work has shown that a single dose of cocaine can produce long-term potentiation (LTP) of the glutamatergic synapses received by dopamine neurons in the ventral tegmental area (VTA). This and other plastic changes in the brain's reward circuitry have been suggested to underlie addiction. A recent study has provided new insights into cocaine-induced LTP, showing that it begins 3-5 h after exposure, requires activation of a dopamine D(5)/NMDA receptor cascade, and can be evoked by cocaine application directly to the VTA. PMID- 19297517 TI - The main sequence of human optokinetic afternystagmus (OKAN). AB - Different types of fast eye movements, including saccades and fast phases of optokinetic nystagmus (OKN) and optokinetic afternystagmus (OKAN), are coded by only partially overlapping neural networks. This is a likely cause for the differences that have been reported for the dynamic parameters of fast eye movements. The dependence of two of these parameters-peak velocity and duration on saccadic amplitude has been termed "main sequence." The main sequence of OKAN fast phases has not yet been analyzed. These eye movements are unique in that they are generated by purely subcortical control mechanisms and that they occur in complete darkness. In this study, we recorded fast phases of OKAN and OKN as well as visually guided and spontaneous saccades under identical background conditions because background characteristics have been reported to influence the main sequence of saccades. Our data clearly show that fast phases of OKAN and OKN differ with respect to their main sequence. OKAN fast phases were characterized by their lower peak velocities and longer durations compared with those of OKN fast phases. Furthermore we found that the main sequence of spontaneous saccades depends heavily on background characteristics, with saccades in darkness being slower and lasting longer. On the contrary, the main sequence of visually guided saccades depended on background characteristics only very slightly. This implies that the existence of a visual saccade target largely cancels out the effect of background luminance. Our data underline the critical role of environmental conditions (light vs. darkness), behavioral tasks (e.g., spontaneous vs. visually guided), and the underlying neural networks for the exact spatiotemporal characteristics of fast eye movements. PMID- 19297518 TI - Factor-analysis methods for higher-performance neural prostheses. AB - Neural prostheses aim to provide treatment options for individuals with nervous system disease or injury. It is necessary, however, to increase the performance of such systems before they can be clinically viable for patients with motor dysfunction. One performance limitation is the presence of correlated trial-to trial variability that can cause neural responses to wax and wane in concert as the subject is, for example, more attentive or more fatigued. If a system does not properly account for this variability, it may mistakenly interpret such variability as an entirely different intention by the subject. We report here the design and characterization of factor-analysis (FA)-based decoding algorithms that can contend with this confound. We characterize the decoders (classifiers) on experimental data where monkeys performed both a real reach task and a prosthetic cursor task while we recorded from 96 electrodes implanted in dorsal premotor cortex. The decoder attempts to infer the underlying factors that comodulate the neurons' responses and can use this information to substantially lower error rates (one of eight reach endpoint predictions) by 150 ms, although still advantageous at shorter timescales, that Gaussian-based algorithms performed better than the analogous Poisson-based algorithms and that the FA algorithm is robust even with a limited amount of training data. We propose that FA-based methods are effective in modeling correlated trial-to-trial neural variability and can be used to substantially increase overall prosthetic system performance. PMID- 19297519 TI - CYP4F2 is a vitamin K1 oxidase: An explanation for altered warfarin dose in carriers of the V433M variant. AB - Genetic polymorphisms in VKORC1 and CYP2C9, genes controlling vitamin K(1) (VK1) epoxide reduction and (S)-warfarin metabolism, respectively, are major contributors to interindividual variability in warfarin dose. The V433M polymorphism (rs2108622) in CYP4F2 has also been associated with warfarin dose and speculatively linked to altered VK1 metabolism. Therefore, the purpose of the present study was to determine the role of CYP4F2 and the V433M polymorphism in the metabolism of VK1 by human liver. In vitro metabolic experiments with accompanying liquid chromatography-tandem mass spectrometry analysis demonstrated that recombinant CYP4F2 (Supersomes) and human liver microsomes supplemented with NADPH converted VK1 to a single product. A screen of all commercially available P450 Supersomes showed that only CYP4F2 was capable of metabolizing VK1 to this product. Steady-state kinetic analysis with recombinant CYP4F2 and with human liver microsomes revealed a substrate K(m) of 8 to 10 microM. Moreover, anti CYP4F2 IgG, as well as several CYP4F2-selective chemical inhibitors, substantially attenuated the microsomal reaction. Finally, human liver microsomes genotyped for rs2108622 demonstrated reduced vitamin K(1) oxidation and lower CYP4F2 protein concentrations in carriers of the 433M minor allele. These data demonstrate that CYP4F2 is a vitamin K(1) oxidase and that carriers of the CYP4F2 V433M allele have a reduced capacity to metabolize VK1, secondary to an rs2108622 dependent decrease in steady-state hepatic concentrations of the enzyme. Therefore, patients with the rs2108622 polymorphism are likely to have elevated hepatic levels of VK1, necessitating a higher warfarin dose to elicit the same anticoagulant response. PMID- 19297520 TI - Pharmacology of vanilloid transient receptor potential cation channels. AB - Depending on their primary structure, the 28 mammalian transient receptor potential (TRP) cation channels identified so far can be sorted into 6 subfamilies: TRPC ("Canonical"), TRPV ("Vanilloid"), TRPM ("Melastatin"), TRPP ("Polycystin"), TRPML ("Mucolipin"), and TRPA ("Ankyrin"). The TRPV subfamily (vanilloid receptors) comprises channels critically involved in nociception and thermosensing (TRPV1, TRPV2, TRPV3, and TRPV4), whereas TRPV5 and TRPV6 are involved in renal Ca(2+) absorption/reabsorption. Apart from TRPV1, the pharmacology of these channels is still insufficiently known. Furthermore, only few small-molecule ligands for non-TRPV1 vanilloid receptors have been identified, and little is known of their endogenous ligands, resulting in a substantial "orphan" state for these channels. In this review, we summarize the pharmacological properties of members of the TRPV subfamily, highlighting the critical issues and challenges facing their "deorphanization" and clinical exploitation. PMID- 19297521 TI - Elucidation of binding sites of dual antagonists in the human chemokine receptors CCR2 and CCR5. AB - Design of dual antagonists for the chemokine receptors CCR2 and CCR5 will be greatly facilitated by knowledge of the structural differences of their binding sites. Thus, we computationally predicted the binding site of the dual CCR2/CCR5 antagonist N-dimethyl-N-[4-[[[2-(4-methylphenyl)-6,7-dihydro-5H-benzohepten-8-yl] carbonyl]amino]benzyl]tetrahydro-2H-pyran-4-aminium (TAK-779), and a CCR2 specific antagonist N-(carbamoylmethyl)-3-trifluoromethyl benzamido parachlorobenzyl 3-aminopyrrolidine (Teijin compound 1) in an ensemble of predicted structures of human CCR2 and CCR5. Based on our predictions of the protein-ligand interactions, we examined the activity of the antagonists for cells expressing thirteen mutants of CCR2 and five mutants of CCR5. The results show that residues Trp98(2.60) and Thr292(7.40) contribute significantly to the efficacy of both TAK-779 and Teijin compound 1, whereas His121(3.33) and Ile263(6.55) contribute significantly only to the antagonistic effect of Teijin compound 1 at CCR2. Mutation of residues Trp86(2.60) and Tyr108(3.32) adversely affected the efficacy of TAK-779 in antagonizing CCR5-mediated chemotaxis. Y49A(1.39) and E291A(7.39) mutants of CCR2 showed a complete loss of CCL2 binding and chemotaxis, despite robust cell surface expression, suggesting that these residues are critical in maintaining the correct receptor architecture. Modeling studies support the hypothesis that the residues Tyr49(1.39), Trp98(2.60), Tyr120(3.32), and Glu291(7.39) of CCR2 form a tight network of aromatic cluster and polar contacts between transmembrane helices 1, 2, 3, and 7. PMID- 19297522 TI - Polarized growth in budding yeast in the absence of a localized formin. AB - Polarity is achieved partly through the localized assembly of the cytoskeleton. During growth in budding yeast, the bud cortex and neck localized formins Bni1p and Bnr1p nucleate and assemble actin cables that extend along the bud-mother axis, providing tracks for secretory vesicle delivery. Localized formins are believed to determine the location and polarity of cables, hence growth. However, yeast expressing the nonlocalized actin nucleating/assembly formin homology (FH) 1-FH2 domains of Bnr1p or Bni1p as the sole formin grow well. Although cables are significantly disorganized, analysis of directed transport of secretory vesicles is still biased toward the bud, reflecting a bias in correctly oriented cables, thereby permitting polarized growth. Myosin II, localized at the bud neck, contributes to polarized growth as a mutant unable to interact with F-actin further compromises growth in cells with an unlocalized formin but not with a localized formin. Our results show that multiple mechanisms contribute to cable orientation and polarized growth, with localized formins and myosin II being two major contributors. PMID- 19297523 TI - Conformational maturation and post-ER multisubunit assembly of gap junction proteins. AB - For all previously well-characterized oligomeric integral membrane proteins, folding, multisubunit assembly, and recognition of conformationally immature molecules for degradation occurs at their organelle of synthesis. This cannot, however, be the case for the gap junction-forming protein connexin43 (Cx43), which when endogenously expressed undergoes multisubunit assembly into connexons only after its transport to the trans-Golgi network. We have developed two novel assays to assess Cx43 folding and assembly: acquisition of resistance of disulfide bonds to reduction by extracellularly added DTT and Triton X-114 detergent phase partitioning. We show that Cx43 synthesized at physiologically relevant levels undergoes a multistep conformational maturation process in which folding of connexin monomers within the ER is a prerequisite for multisubunit assembly in the TGN. Similar results were obtained with Cx32, disproving the widely reported contention that the site of endogenous beta connexin assembly is the ER. Exogenous overexpression of Cx43, Cx32, or Cx26 allows these events to take place within the ER, the first example of the TGN and ER as alternative sites for oligomeric assembly. Our findings also constitute the first biochemical evidence that defective connexin folding is a cause of the human disorder X linked Charcot-Marie-Tooth disease. PMID- 19297524 TI - Role of p54 RNA helicase activity and its C-terminal domain in translational repression, P-body localization and assembly. AB - The RNA helicase p54 (DDX6, Dhh1, Me31B, Cgh-1, RCK) is a prototypic component of P-(rocessing) bodies in cells ranging from yeast to human. Previously, we have shown that it is also a component of the large cytoplasmic polyadenylation element-binding protein translation repressor complex in Xenopus oocytes and that when tethered to the 3' untranslated region, Xp54 represses reporter mRNA translation. Here, we examine the role of the p54 helicase activity in translational repression and in P-body formation. Mutagenesis of conserved p54 helicase motifs activates translation in the tethered function assay, reduces accumulation of p54 in P-bodies in HeLa cells, and inhibits its capacity to assemble P-bodies in p54-depleted cells. Similar results were obtained in four helicase motifs implicated in ATP binding and in coupling ATPase and RNA binding activities. This is accompanied by changes in the interaction of the mutant p54 with the oocyte repressor complex components. Surprisingly, the C-terminal D2 domain alone is sufficient for translational repression and complete accumulation in P-bodies, although it is deficient for P-body assembly. We propose a novel RNA helicase model, in which the D2 domain acts as a protein binding platform and the ATPase/helicase activity allows protein complex remodeling that dictates the balance between repressors and an activator of translation. PMID- 19297525 TI - Identification of the signal directing Tim9 and Tim10 into the intermembrane space of mitochondria. AB - The intermembrane space of mitochondria contains the specific mitochondrial intermembrane space assembly (MIA) machinery that operates in the biogenesis pathway of precursor proteins destined to this compartment. The Mia40 component of the MIA pathway functions as a receptor and binds incoming precursors, forming an essential early intermediate in the biogenesis of intermembrane space proteins. The elements that are crucial for the association of the intermembrane space precursors with Mia40 have not been determined. In this study, we found that a region within the Tim9 and Tim10 precursors, consisting of only nine amino acid residues, functions as a signal for the engagement of substrate proteins with the Mia40 receptor. Furthermore, the signal contains sufficient information to facilitate the transfer of proteins across the outer membrane to the intermembrane space. Thus, here we have identified the mitochondrial intermembrane space sorting signal required for delivery of proteins to the mitochondrial intermembrane space. PMID- 19297526 TI - Tetrahymena meiotic nuclear reorganization is induced by a checkpoint kinase dependent response to DNA damage. AB - In the ciliate Tetrahymena, meiotic micronuclei (MICs) undergo extreme elongation, and meiotic pairing and recombination take place within these elongated nuclei (the "crescents"). We have previously shown that elongation does not occur in the absence of Spo11p-induced DNA double-strand breaks (DSBs). Here we show that elongation is restored in spo11Delta mutants by various DNA-damaging agents including ones that may not cause DSBs to a notable extent. MIC elongation following Spo11p-induced DSBs or artificially induced DNA lesions is probably a DNA-damage response mediated by a phosphokinase signal transduction pathway, since it is suppressed by the ATM/ATR kinase inhibitors caffeine and wortmannin and by knocking out Tetrahymena's ATR orthologue. MIC elongation occurs concomitantly with the movement of centromeres away from the telomeric pole of the MIC. This DNA damage-dependent reorganization of the MIC helps to arrange homologous chromosomes alongside each other but is not sufficient for exact pairing. Thus, Spo11p contributes to bivalent formation in two ways: by creating a favorable spatial disposition of homologues and by stabilizing pairing by crossovers. The polarized chromosome orientation inside the crescent resembles the conserved meiotic bouquet, and crescent and bouquet also share the putative function of aiding meiotic pairing. However, they are regulated differently because in Tetrahymena, DSBs are required for entering rather than exiting this stage. PMID- 19297527 TI - Prm3p is a pheromone-induced peripheral nuclear envelope protein required for yeast nuclear fusion. AB - Nuclear membrane fusion is the last step in the mating pathway of the yeast Saccharomyces cerevisiae. We adapted a bioinformatics approach to identify putative pheromone-induced membrane proteins potentially required for nuclear membrane fusion. One protein, Prm3p, was found to be required for nuclear membrane fusion; disruption of PRM3 caused a strong bilateral defect, in which nuclear congression was completed but fusion did not occur. Prm3p was localized to the nuclear envelope in pheromone-responding cells, with significant colocalization with the spindle pole body in zygotes. A previous report, using a truncated protein, claimed that Prm3p is localized to the inner nuclear envelope. Based on biochemistry, immunoelectron microscopy and live cell microscopy, we find that functional Prm3p is a peripheral membrane protein exposed on the cytoplasmic face of the outer nuclear envelope. In support of this, mutations in a putative nuclear localization sequence had no effect on full-length protein function or localization. In contrast, point mutations and deletions in the highly conserved hydrophobic carboxy-terminal domain disrupted both protein function and localization. Genetic analysis, colocalization, and biochemical experiments indicate that Prm3p interacts directly with Kar5p, suggesting that nuclear membrane fusion is mediated by a protein complex. PMID- 19297529 TI - The exocyst protein Sec10 is necessary for primary ciliogenesis and cystogenesis in vitro. AB - Primary cilia are found on many epithelial cell types, including renal tubular epithelial cells, in which they are felt to participate in flow sensing and have been linked to the pathogenesis of cystic renal disorders such as autosomal dominant polycystic kidney disease. We previously localized the exocyst, an eight protein complex involved in membrane trafficking, to the primary cilium of Madin Darby canine kidney cells and showed that it was involved in cystogenesis. Here, using short hairpin RNA (shRNA) to knockdown exocyst expression and stable transfection to induce exocyst overexpression, we show that the exocyst protein Sec10 regulates primary ciliogenesis. Using immunofluorescence, scanning, and transmission electron microscopy, primary cilia containing only basal bodies are seen in the Sec10 knockdown cells, and increased ciliogenesis is seen in Sec10 overexpressing cells. These phenotypes do not seem to be because of gross changes in cell polarity, as apical, basolateral, and tight junction proteins remain properly localized. Sec10 knockdown prevents normal cyst morphogenesis when the cells are grown in a collagen matrix, whereas Sec10 overexpression results in increased cystogenesis. Transfection with human Sec10 resistant to the canine shRNA rescues the phenotype, demonstrating specificity. Finally, Par3 was recently shown to regulate primary cilia biogenesis. Par3 and the exocyst colocalized by immunofluorescence and coimmunoprecipitation, consistent with a role for the exocyst in targeting and docking vesicles carrying proteins necessary for primary ciliogenesis. PMID- 19297528 TI - Down-regulation of the met receptor tyrosine kinase by presenilin-dependent regulated intramembrane proteolysis. AB - Hepatocyte growth factor/scatter factor (HGF/SF) acts through the membrane anchored Met receptor tyrosine kinase to induce invasive growth. Deregulation of this signaling is associated with tumorigenesis and involves, in most cases, overexpression of the receptor. We demonstrate that Met is processed in epithelial cells by presenilin-dependent regulated intramembrane proteolysis (PS RIP) independently of ligand stimulation. The proteolytic process involves sequential cleavage by metalloproteases and the gamma-secretase complex, leading to generation of labile fragments. In normal epithelial cells, although expression of cleavable Met by PS-RIP is down-regulated, uncleavable Met displayed membrane accumulation and induced ligand-independent motility and morphogenesis. Inversely, in transformed cells, the Met inhibitory antibody DN30 is able to promote Met PS-RIP, resulting in down-regulation of the receptor and inhibition of the Met-dependent invasive growth. This demonstrates the original involvement of a proteolytic process in degradation of the Met receptor implicated in negative regulation of invasive growth. PMID- 19297530 TI - Liver progenitor cells fold up a cell monolayer into a double-layered structure during tubular morphogenesis. AB - Bile ducts are hepatic tubular structures that are lined by cholangiocytes, a type of liver epithelial cell. Cholangiocytes first form a single layer of cells, termed the ductal plate, surrounding the portal vein, which eventually remodels into the branching tubular network of bile ducts. The process of bile duct morphogenesis is not yet clear: a conventional model where cholangiocytes proliferate to duplicate a single layer of the ductal plate before lumen formation seems inconsistent with the observation that proliferation is dramatically reduced when hepatoblasts, liver progenitor cells, differentiate into cholangiocytes. Here, we developed a new culture system in which a liver progenitor cell line, HPPL, reorganizes from a monolayer to tubular structures in response to being overlaid with a gel containing type I collagen and Matrigel. We found that some of the HPPL in the monolayer depolarized and migrated to fold up the monolayer into a double-cell layer. These morphogenetic processes occurred without cell proliferation and required phosphatidylinositol 3-kinase and Akt activity. Later in morphogenesis, luminal space was generated between the two cell layers. This process, in particular enlargement of the apical lumen, involved transcriptional activity of HNF1beta. Thus, using this sandwich culture system, we could segregate tubulogenesis of bile ducts into distinct steps and found that the PI3K/Akt pathway and HNF1beta regulated different steps of the morphogenesis. Although the process of tubulogenesis in culture specifically resembled early bile duct formation, involvement of these two key players suggests that the sandwich culture might help us to find common principles of tubulogenesis in general. PMID- 19297531 TI - Focal adhesion kinase modulates cell adhesion strengthening via integrin activation. AB - Focal adhesion kinase (FAK) is an essential nonreceptor tyrosine kinase regulating cell migration, adhesive signaling, and mechanosensing. Using FAK-null cells expressing FAK under an inducible promoter, we demonstrate that FAK regulates the time-dependent generation of adhesive forces. During the early stages of adhesion, FAK expression in FAK-null cells enhances integrin activation to promote integrin binding and, hence, the adhesion strengthening rate. Importantly, FAK expression regulated integrin activation, and talin was required for the FAK-dependent effects. A role for FAK in integrin activation was confirmed in human fibroblasts with knocked-down FAK expression. The FAK autophosphorylation Y397 site was required for the enhancements in adhesion strengthening and integrin-binding responses. This work demonstrates a novel role for FAK in integrin activation and the time-dependent generation of cell-ECM forces. PMID- 19297532 TI - Uranium isotopic ratio determination in urine using flow-injection ICP-MS: a tool for emergency monitoring. AB - An inductively coupled plasma mass spectrometry-based method is presented aimed at total uranium concentration and atomic ratio determination. The method includes flow-injection uranium separation based on TRU Eichrom extraction chromatographic cartridges. The method was tested with urine interlaboratorial exercise samples and certified reference materials (NBL-CRM-U020A and NBL-CRM U050) providing reliable results. The proposed methodology was also applied to urine samples obtained after an incident at a nuclear facility. The obtained (235)U/(238)U and (234)U/(238)U atomic ratios in the urine of an exposed worker were equal to those observed for an enriched uranium solution from the same origin. PMID- 19297533 TI - New findings: a very high natural radiation area in Afra hot springs, Jordan. AB - A high natural radiation zone was investigated for the first time in Afra hot springs of Jordan. The radiation levels were measured using a portable Geiger Muller counter and an Na(Tl) detector. The measured absorbed dose rates in air ranged from 10 to 1800 nGy h(-1), suggesting that the concentration of natural radioactive materials is very high compared with their normal abundance in crustal rocks. A single high-radiation zone was also found in a nearby area where a gamma radiation dose rate of 4.0 mGy h(-1) was measured. On the basis of this measurement, the area was marked as a high-radiation zone. This region is far from tourist areas and not easily reached. No intervention measures are needed to protect people because the spa area is not well inhabited, having only daily visitors (average frequency of 10 days per year per individual). The dose received by workers in the spa area should be considered and the worker should be monitored by personal radiation dosimeters, such as thermoluminescent dosimeters. PMID- 19297534 TI - Natural radioactivity of 226Ra and 228Ra in thermal and mineral waters in Croatia. AB - Thermal waters are known as valuable natural resources of a country. They contain certain degree of natural radioactivity attributable to the elements of the uranium and thorium natural decay series. Among these elements, the most radiotoxic and the most important is radium that exists in several isotopic forms (226Ra and 228Ra). The focus of attention was the content of radium in samples of thermal and mineral spring water from several spas in Croatia. These waters are mainly used for medical, bathing and recreational purposes, and some of them are used for drinking. Measured activity concentrations of 226Ra ranged from 87 to 6200 mBq l(-1) which, in some springs, exceed the maximal permissible level of 1 Bq l(-1) for drinking water. Measured activity concentrations of 228Ra ranged from 23 to 3480 mBq l(-1). The study showed that radium content for the investigated thermal and mineral waters is below the levels at which negative consequences would arise due to ingestion. PMID- 19297535 TI - Honor, brotherhood, and the corporate ethos of London's Barber-Surgeons' Company, 1570-1640. AB - As the largest and most civically active body of medical practitioners in the late Tudor and early Stuart period, surgeons played a vital role in London's urban landscape, but remained precariously vulnerable to abasement due to the regular contact with death and disease necessitated by their work. Based on an analysis of guild records, printed surgical manuals, and conduct literature, this study explores the emergent corporate ethos of London's Barber-Surgeons' Company and addresses the identity formation of surgeons in the late-sixteenth and early seventeenth centuries. By implementing codes of conduct and uniform standards of practice, punishing transgressions of propriety, and developing legislation to limit the activities of unlicensed and foreign practitioners, Company officers ardently sought social and occupational legitimacy within a milieu characterized by a tremendous emphasis on status and hierarchy. Rooted in methodology drawn from the social history of medicine and cultural anthropology, this study argues that in response to the persistent stigma associated with their work and London's increasingly prevalent culture of credit, surgeons, like other artisanal groups, sought to enhance their social legitimacy and occupational respectability by manipulating contemporary social rituals, reinforcing the honorable associations of their work, and preserving the veneer of brotherhood and camaraderie. PMID- 19297536 TI - Humanized-monoclonal antibody against heterologous Leptospira infection. AB - Patients with leptospirosis are commonly treated with antibiotics. Jarisch Herxheimer reaction caused by toxic bacterial substances massively released as a result of the antibiotic mediated-bacterial lysis occurs in some patients which may aggravate the existing severe clinical manifestations. In this study, a humanized-murine single-chain monoclonal antibody (HuScFv) was produced and tested as an alternative of antibiotics for treatment of leptospirosis. Complementary DNA was prepared from total RNA of a murine hybridoma clone secreting monoclonal antibody (MAb) specific to LipL32 of pathogenic Leptospira spp. The MAb had therapeutic efficacy in Leptospira challenged hamsters. The VH and VL coding sequences were amplified using the cDNA as a template. The sequences were linked to form a single-chain variable murine DNA fragment (muscFv). CDR sequences of the muscFv were grafted onto the best matching human VH and VL immunoglobulin frameworks. After cloning of the humanized murine DNA sequences (huscFv) into a phagemid vector and the vector was introduced into competent Escherichia coli, the HuScFv was produced. On the same weight basis, the HuScFv possessed equal neutralizing activities to the murine ScFv counterpart against heterologous Leptospira-mediated hemolysis in vitro and rescued hamsters from a heterologous Leptospira lethal challenge. The HuScFv antibody has high therapeutic potential as an alternative to antibiotics for human leptospirosis, especially for drug hypersensitive patients. PMID- 19297537 TI - Blockade of PGHS-2 inhibits the hypothalamus-pituitary-adrenal axis response to cerebral hypoperfusion in the sheep fetus. AB - Decreases in fetal blood pressure stimulate homeostatic stress responses that help return blood pressure to normal levels. Fetal hypothalamus-pituitary-adrenal (HPA) axis responses to hypotension are mediated by chemoreceptor and baroreceptor reflexes and ischemia of the fetal central nervous system. Indomethacin, a nonselective inhibitor of prostaglandin endoperoxide synthase (PGHS)-1 and -2, attenuates the HPA response to hypotension in the fetus. The present study was designed to test the hypothesis that selective inhibition of PGHS-2 also inhibits the HPA response to cerebral hypoperfusion. We studied 13 chronically catheterized fetal sheep (126-136 days gestation). Five fetal sheep were subjected to intracerebroventricular infusion of nimesulide (0.01 mg/day), a specific inhibitor of PGHS-2, and eight were treated with vehicle (DMSO in water) for 5 days. Each fetus was subjected to a 10-min period of brachiocephalic occlusion, which decreased carotid arterial pressure approximately 75% and reflexively increased fetal plasma concentrations of ACTH, POMC, cortisol, and femoral arterial pressure, and decreased fetal heart rate. Nimesulide significantly inhibited the ACTH response to the BCO, while significantly augmenting the reflex cardiovascular response and altering fetal heart rate variability consistent with increased sympathetic nervous system activity. The results of this study demonstrate that the activity of PGHS-2 in the brain is a necessary component of the fetal HPA response to cerebral hypoperfusion in the late-gestation fetal sheep. These results are consistent with those of recent study, in which we demonstrated that the preparturient increase in fetal ACTH secretion depends upon PGHS-2 activity within the fetal brain. PMID- 19297538 TI - Phenobarbital blockade of the preovulatory luteinizing hormone surge: association with phase-advanced circadian clock and altered suprachiasmatic nucleus Period1 gene expression. AB - The suprachiasmatic nucleus (SCN) controls the timing of the preovulatory luteinizing hormone (LH) surge in laboratory rodents. Barbiturate administration during a critical period on proestrus delays the surge and prolongs the estrous cycle 1 day. Because a nonphotic timing signal (zeitgeber) during the critical period that phase advances activity rhythms can also induce the latter effect, we hypothesized that barbiturates delay the LH surge by phase-advancing its circadian timing signal beyond the critical period. In experiment 1, locomotor rhythms and estrous cycles were monitored in hamsters for 2-3 wk preinjection and postinjection of vehicle or phenobarbital and after transfer to darkness at zeitgeber time (ZT) 6 on proestrus. Phenobarbital delayed estrous cycles in five of seven hamsters, which exhibited phase shifts that averaged twofold greater than those exhibited by vehicle controls or phenobarbital-injected hamsters with normal cycles. Experiment 2 used a similar protocol, but injections were at ZT 5, and blood samples for LH determination were collected from 1200 to 1800 on proestrus and the next day via jugular cannulae inserted the day before proestrus. Phenobarbital delayed the LH surge 1 day in all six hamsters, but it occurred at an earlier circadian time, supporting the above hypothesis. Experiment 3 investigated whether phenobarbital, like other nonphotic zeitgebers, suppresses SCN Period1 and Period2 transcription. Two hours postinjection, phenobarbital decreased SCN expression of only Period1 mRNA, as determined by in situ hybridization. These results suggest that phenobarbital advances the SCN pacemaker, governing activity rhythms and hormone release in part by decreasing its Period1 gene expression. PMID- 19297539 TI - Cold stimulates the behavioral response to hypoxia in newborn mice. AB - In newborns, hypoxia elicits increased ventilation, arousal followed by defensive movements, and cries. Cold is known to affect the ventilatory response to hypoxia, but whether it affects the arousal response remains unknown. The aim of the present study was to assess the effects of cold on the ventilatory and arousal responses to hypoxia in newborn mice. We designed an original platform measuring noninvasively and simultaneously the breathing pattern by whole body plethysmography, body temperature by infrared thermography, as well as motor and ultrasonic vocal (USV) responses. Six-day-old mice were exposed twice to 10% O(2) for 3 min at either cold temperature (26 degrees C) or thermoneutrality (33 degrees C). At 33 degrees C, hypoxia elicited a marked increase in ventilation followed by a small ventilatory decline, small motor response, and almost no USVs. Body temperature was not influenced by hypoxia, and oxygen consumption (Vo(2)) displayed minimal changes. At 26 degrees C, hypoxia elicited a slight increase in ventilation with a large ventilatory decline and a large drop of Vo(2). This response was accompanied by marked USV and motor responses. Hypoxia elicited a small decrease in temperature after the return to normoxia, thus precluding any causal influence on the motor and USV responses to hypoxia. In conclusion, cold stimulated arousal and stress responses to hypoxia, while depressing hypoxic hyperpnea. Arousal is an important defense mechanism against sleep-disordered breathing. The dissociation between ventilatory and behavioral responses to hypoxia suggests that deficits in the arousal response associated with sleep breathing disorders cannot be attributed to a depressed hypoxic response. PMID- 19297540 TI - Microinjections of alpha-melanocyte stimulating hormone into the nucleus ambiguus of the rat elicit vagally mediated bradycardia. AB - Neurons that immunostain for alpha-melanocyte stimulating hormone (alpha-MSH) have been identified in the nucleus ambiguus (nAmb). The presence of mRNA for melanocortin type 4 receptors (MC4Rs) has also been reported in this nucleus. On the basis of this information, it was hypothesized that activation of MC4Rs in the nAmb may play a role in the regulation of cardiac function. This hypothesis was tested in urethane-anesthetized, artificially ventilated, adult male Wistar rats. Microinjections (30 nl) of alpha-MSH (0.1, 0.2, 0.4, 0.8, and 1.2 mM) into the nAmb of anesthetized rats elicited decreases in heart rate (HR; 1.3 +/- 0.6, 3 +/- 1, 11 +/- 2, 46.3 +/- 3, and 43.3 +/- 7 bpm, respectively) and no changes in mean arterial pressure (MAP). Maximum decreases in HR were elicited by 0.8 mM concentration of alpha-MSH. Bradycardic responses to alpha-MSH were similar in unanesthetized midcollicular decerebrate rats. Microinjections of artificial cerebrospinal fluid (30 nl) into the nAmb did not elicit a HR response. Bilateral vagotomy completely abolished alpha-MSH-induced bradycardia. The decreases in HR elicited by alpha-MSH (0.8 mM) were completely blocked by a selective MC4R antagonist. Direct application of alpha-MSH on the nAmb neurons increased their firing, which was blocked by prior applications of the MC4R antagonist. Microinjections of the MC4R antagonist into the nAmb did not alter reflex bradycardic responses elicited by intravenous infusions of phenylephrine, suggesting that MC4Rs did not play a role in mediating the parasympathetic component of baroreflex-induced bradycardia. These results indicated that alpha MSH microinjections into the nAmb exert excitatory effects on parasympathetic preganglionic nAmb neurons via MC4Rs, leading to bradycardic responses. PMID- 19297541 TI - Sympathetic restraint of muscle blood flow during hypoxic exercise. AB - Control of exercising muscle blood flow is a balance between local vasodilatory factors and the increase in global sympathetic vasoconstrictor outflow. Hypoxia has been shown to potentiate the muscle sympathetic nerve response to exercise, potentially limiting the increase in muscle blood flow. Accordingly, we investigated sympathetic restraint to exercising muscle during whole body exercise in hypoxia. Six dogs chronically instrumented with ascending aortic and hindlimb flow probes and a terminal aortic catheter were studied at rest and mild [2.5 miles/h (mph), 5% grade] and moderate (4.0 mph, 10% grade) exercise while breathing room air or hypoxia (Pa(O(2)) approximately 45 mmHg) in the intact control condition and following systemic alpha-adrenergic blockade (phentolamine). Hypoxia caused an increase in cardiac output (CO), hindlimb flow (Flow(L)), and blood pressure (BP), while total (Cond(T)) and hindlimb conductance (Cond(L)) were unchanged at rest and mild exercise but increased with moderate exercise. During both mild and moderate exercise, alpha-blockade in normoxia resulted in significant vasodilation as evidenced by increases in CO (10%), Flow(L) (17%), Cond(T) (33%), Cond(L) (43%), and a decrease in BP (-18%), with the increase in Cond(L) greater than the increase in Cond(T) during mild exercise. Compared with the normoxic response, alpha-blockade in hypoxia during exercise resulted in a significantly greater increase in Cond(T) (59%) and Cond(L) (74%) and a correspondingly greater decrease in BP (-34%) from baseline. These findings indicate that there is considerable hypoxia-induced sympathetic restraint of muscle blood flow during both mild and moderate exercise, which helps to maintain arterial blood pressure in hypoxia. PMID- 19297542 TI - Drinking-induced thermoregulatory panting in rehydrated sheep: influences of oropharyngeal/esophageal signals, core temperature, and thirst satiety. AB - Dehydrated mammals conserve body water by reducing thermoregulatory evaporative cooling responses e.g., panting and sweating. Increased core temperature (Tc) may result. Following rehydration and correction of fluid deficits, panting and sweating commence. We investigated the role of oropharyngeal/esophageal, postabsorptive and thermal signals in the panting response, and reduced Tc that occurs when unshorn sheep drink water following water deprivation for 2 days (ambient temperature 20 degrees C). Ingestion of water (at body temperature) resulted in increased respiratory rate (panting) and reduced Tc within 4 min that persisted for at least 90 min. Initially, a similar panting response and reduced Tc occurred following rehydration by drinking isotonic saline solution, but panting was not sustained after 20 min, and Tc began to rise again. Rehydration by intraruminal administration of water, without any drinking, resulted in delayed panting and fall in Tc. Intraruminal infusion of saline was ineffective. Rehydration by drinking cool water (20 degrees C) resulted in a rapid fall in Tc without increased panting. Shorn sheep had lower basal Tc that did not increase during 2 days of water deprivation, and they did not pant on rehydration by drinking water. Our results indicate that signals from the oropharyngeal and/or esophageal region associated with the act of drinking play a crucial role in the initial 20-30 min of the panting response to rehydration. Postabsorptive factors most likely reduced plasma tonicity and cause continued panting and further reduction in Tc. Tc also influences rehydration-induced panting. It occurs only if sheep incur a heat load during bodily dehydration. PMID- 19297543 TI - Photoperiod and acute energy deficits interact on components of the thyroid hormone system in hypothalamic tanycytes of the Siberian hamster. AB - In the Siberian hamster, seasonal weight loss occurs gradually over many weeks during autumn and winter. This is driven by a regulatory mechanism that is able to integrate duration of exposure to short days (SDs) with the size of body energy reserves. After food restriction in SDs, followed by ad libitum refeeding, body weight of the hamster does not return to its former level; rather, it increases to a level defined by the length of time spent in SDs. In this report, we show that components of the thyroid hormone system that are involved in seasonal weight loss change expression in response to 48 h of starvation. Eight weeks in an SD photoperiod induced weight loss in the Siberian hamster. In the hypothalamus of these hamsters, type II deiodinase expression was decreased and type III deiodinase expression was induced, but there was no change in hypothalamic neuropeptide Y or thyrotropin-releasing hormone gene expression. For the first time, we show that the thyroid hormone transporter monocarboxylate transporter 8 is expressed in tanycytes and is increased in response to an SD photoperiod. Food restriction (48 h of starvation) reversed the direction of gene expression change for type II and III deiodinase and monocarboxylate transporter 8 induced by SD photoperiods. Furthermore, fasting increased neuropeptide Y expression and decreased thyrotropin-releasing hormone expression. VGF, a gene upregulated in SDs in the dorsal region of the medial posterior area of the arcuate nucleus, was not changed by starvation. These data point to a mechanism whereby energy deprivation can interact with an SD photoperiod on hypothalamic tanycytes to regulate components of the thyroid hormone system involved in photoperiodic regulation of seasonal physiology. PMID- 19297544 TI - A high-fat diet attenuates the central response to within-meal satiation signals and modifies the receptor expression of vagal afferents in mice. AB - During digestion, macronutrients are sensed within the small intestine. This sensory process is dependent upon the action of gut mediators, such as cholecystokinin (CCK) or serotonin (5-HT), on vagal afferents that, in turn, convey peripheral information to the brain to influence the control of food intake. Recent studies have suggested that dietary conditions alter vagal sensitivity to CCK and 5-HT. This phenomenon may be of importance to the onset of eating disorders. The aim of the present study was thus to investigate the effects of subjecting mice to 15 days of either an HF diet (30% fat, 54% carbohydrate) or an NF diet (10% fat, 74% carbohydrate) on 1) daily and short term food intake, 2) vagal sensitivity to peripheral anorectic factors and macronutrient loads, and 3) vagal afferent neuron receptor expression. The results indicated that compared with an NF diet, and while increasing food intake and body weight gain, an HF diet altered the short-term response to CCK-8 and intragastric macronutrient loads, while decreasing vagal activation by CCK-8 and modifying the receptor expression of vagal neurons. These findings, therefore, suggest that dietary intervention effect on food intake could be linked to changes in vagal afferent receptor profiles. PMID- 19297545 TI - Effects of aerobic and anaerobic metabolic inhibitors on avian intrapulmonary chemoreceptors. AB - Birds have rapidly responding respiratory chemoreceptors [intrapulmonary chemoreceptors (IPC)] that provide vagal sensory feedback about breathing pattern. IPC are exquisitely sensitive to CO(2) but are unaffected by hypoxia. IPC continue to respond to CO(2) during hypoxic and even anoxic conditions, suggesting that they may generate ATP needed for signal transduction anaerobically. To assess IPC energy metabolism, single-cell action potential discharge and acid-base status were recorded from 26 pentobarbital-anesthetized Anas platyrhynchos before and after intravenous infusion of the glycolytic blocker iodoacetate (10-70 mg/kg), mitochondrial blocker rotenone (2 mg/kg), and/or mitochondrial uncoupler 2,4-dinitrophenol (5-15 mg/kg). After 5 min exposure at the highest dosages, iodoacetate inhibited IPC discharge 65% (15.9 +/ 0.3 s(-1) to 5.5 +/- 0.3 s(-1), P < 0.05), rotenone inhibited discharge 80% (12.9 +/- 0.5 s(-1) to 2.6 +/- 0.6 s(-1), P < 0.05), and 2,4-dinitrophenol inhibited discharge 19% (14.0 +/- 0.3 s(-1) to 11.3 +/- 0.3 s(-1), P < 0.05). These results suggest that IPC utilize glucose, require an intact glycolytic pathway, and metabolize the products of glycolysis to CO(2) and H(2)O by mitochondrial respiration. The small but significant effect of 2,4-dinitrophenol suggests that ATP production by glycolysis may be sufficient to meet IPC energy demands if NADH can be oxidized to NAD experimentally by uncoupling mitochondria, or physiologically by transient lactate production. A model for IPC spike frequency adaptation is proposed, whereby the rapid onset of phasic IPC discharge requires ATP from anaerobic glycolysis, using lactate as the electron acceptor, and the roll-off in IPC discharge reflects transient acidosis due to intracellular lactic acid accumulation. PMID- 19297547 TI - Cell proliferation of human ovarian cancer is regulated by the opioid growth factor-opioid growth factor receptor axis. AB - Ovarian cancer is the leading cause of death from gynecological malignancies. Understanding the biology of these tumors, as well as treatment modalities, has been challenging. The opioid growth factor (OGF; [Met(5)]-enkephalin) and the OGF receptor (OGFr) form an endogenous growth-regulating pathway in homeostasis and neoplasia. In this investigation, we examined the relationship of the OGF-OGFr axis to ovarian cancer, and defined its presence, function, and mechanisms. Using OVCAR-3 and SKOV-3 ovarian cancer cell lines, we found that OGF and OGFr were present and functional. Exogenous OGF was observed to have a dose-dependent, serum-independent, reversible, and receptor-mediated inhibitory action on cell proliferation that was dependent on RNA and protein synthesis. The repressive effect of OGF on cell proliferation also was observed in SW626, CAOV-3, and HEY ovarian cancer cell lines. Endogenous OGF was found to be constitutively produced and tonically active on cell replicative activities, with neutralization of this peptide accelerating cell proliferation. Silencing of OGFr using siRNA technology stimulated cell replication, documenting its integral role. The mechanism of OGF OGFr action on DNA synthesis was related to the cyclin-dependent kinase inhibitory pathway because knockdown of p16 or p21 in OVCAR-3 cells, and p21 in SKOV-3 cells, eliminated OGF's inhibitory effect on growth. These data are the first to report that the OGF-OGFr system is a native biological regulator of cell proliferation in human ovarian cancer. This information will be important in designing treatment strategies for this deadly disease. PMID- 19297546 TI - Exercise training inducibility of MnSOD protein expression and activity is retained while reducing prooxidant signaling in the heart of senescent rats. AB - While the stress response to heat and exercise is limited in the heart with progressive aging, recent data indicate that acute or short-term exercise upregulates the Mn isoform of superoxide dismutase (MnSOD), which may provide protection against ischemia-reperfusion injury and cell death by reducing oxidative stress. Growing evidence indicates that inducible nitric oxide synthase (iNOS) contributes to age-induced increases in oxidative stress and risk of heart failure. We postulated that oxidative stress and iNOS levels would be related to the ability of the aging heart to upregulate MnSOD in response to long-term exercise training. Six- and twenty-seven-mo-old Fischer-344 rats had been assigned to young sedentary (YS), young exercise (YE), old sedentary (OS), or old exercise (OE) groups. ET groups ran on a treadmill for 60 min/day, 5 days/wk for a total of 12 wk. MnSOD protein expression in the left ventricle was increased (+43%) by 12 wk of exercise training in the old age group, with no changes in Cu,ZnSOD. Exercise training also increased MnSOD activity in left ventricles from old and young rats. HSP70 was inducible by exercise training in hearts exclusively from the young age group. iNOS protein expression increased markedly with aging (+548%), while exercise training decreased iNOS levels by -73% in OE compared with OS. In addition, 4-hydroxynonenal protein adducts in the left ventricle increased by 237% with aging, while 12 wk of exercise training resulted in attenuation (-55%). These data indicate that inducibility of MnSOD is preserved with long-term exercise training in the aging rat heart. Moreover, upregulation of MnSOD in the aging heart was directly associated with attenuated levels of oxidative stress, including iNOS. PMID- 19297548 TI - Progressive polyuria without vasopressin neuron loss in a mouse model for familial neurohypophysial diabetes insipidus. AB - Familial neurohypophysial diabetes insipidus (FNDI), an autosomal dominant disorder, is mostly caused by mutations in the gene of neurophysin II (NPII), the carrier protein of arginine vasopressin (AVP). Previous studies suggest that loss of AVP neurons might be the cause of polyuria in FNDI. Here we analyzed knockin mice expressing mutant NPII that causes FNDI in humans. The heterozygous mice manifested progressive polyuria as do patients with FNDI. Immunohistochemical analyses revealed that inclusion bodies that were not immunostained with antibodies for mutant NPII, normal NPII, or AVP were present in the AVP cells in the supraoptic nucleus (SON), and that the size of inclusion bodies gradually increased in parallel with the increases in urine volume. Electron microscopic analyses showed that aggregates existed in the endoplasmic reticulum (ER) as well as in the nucleus of AVP neurons in 1-mo-old heterozygous mice. At 12 mo, dilated ER filled with aggregates occupied the cytoplasm of AVP cells, while few aggregates were found in the nucleus. Analyses with in situ hybridization revealed that expression of AVP mRNA was significantly decreased in the SON in the heterozygous mice compared with that in wild-type mice. Counting cells expressing AVP mRNA in the SON indicated that polyuria had progressed substantially in the absence of neuronal loss. These data suggest that cell death is not the primary cause of polyuria in FNDI, and that the aggregates accumulated in the ER might be involved in the dysfunction of AVP neurons that lead to the progressive polyuria. PMID- 19297549 TI - Extracellular invertase LIN6 of tomato: a pivotal enzyme for integration of metabolic, hormonal, and stress signals is regulated by a diurnal rhythm. AB - To elucidate the regulation pattern of extracellular invertase LIN6 of tomato, the corresponding promoter has been cloned and the sink-tissue specific expression and its regulation by sugars, stress stimuli, growth regulators, and the diurnal rhythm is shown. The in situ analysis of transgenic tobacco plants expressing a LIN6 promoter::beta-glucuronidase reporter gene fusion demonstrates LIN6 expression in sink tissues, such as pollen grains and vascular tissues of leaves and stems. LIN6 is up-regulated in close proximity to wounded tissue, and by methyl jasmonate and abscisic acid, global signals known to modulate defence/stress response. Salicylic acid on the other hand, as well as acetyl salicylic acid, suppresses LIN6 expression, supporting the fact that LIN6 is an inducible compound of the defence/stress response pathway that is antagonistically regulated by jasmonates and salicylates. Induction of the LIN6 promoter in stable transformed BY2 suspension cultures by sucrose and the growth promoting phytohormones cytokinin and auxin along histochemical expression data, showing LIN6 expression in germinating seeds and seedlings, indicates a role of LIN6 invertase during growth processes. In addition, LIN6 is regulated by a diurnal rhythm that drives LIN6 expression in subjective dawn. Transactivation assays with circadian oscillator elements of Arabidopsis Circadian Clock Associated 1 and Late Elongated Hypocotyl demonstrate functional interaction with the LIN6 promoter. PMID- 19297550 TI - Darwin's foundation for investigating self-incompatibility and the progress toward a physiological model for S-RNase-based SI. AB - Charles Darwin made extensive observations of the pollination biology of a wide variety of plants. He carefully documented the consequences of self-pollination and described species that were self-sterile but that could easily be crossed with other plants of the same species. He believed that compatibility was controlled by the 'mutual action' of pollen and pistil contents. A genetic model for self-sterility was developed in the early 1900 s based on studies of the compatibility relationships among, what are now referred to as, self-incompatible (SI) Nicotiana species. Today, it is believed that SI in these species is controlled by an interaction between S-RNases produced in the pistil and F-box proteins expressed in pollen and, moreover, that this S-RNase-based SI system is shared by a great diversity of other plant species. Current research is aimed at understanding how the mutual actions of these S-gene products function in the physiological context of pollen tube growth. PMID- 19297551 TI - Length and breadth of ubiquitin: development to defence in plant life cycle. Preface. PMID- 19297552 TI - A roadmap for zinc trafficking in the developing barley grain based on laser capture microdissection and gene expression profiling. AB - Nutrients destined for the developing cereal grain encounter several restricting barriers on their path towards their final storage sites in the grain. In order to identify transporters and chelating agents that may be involved in transport and deposition of zinc in the barley grain, expression profiles have been generated of four different tissue types: the transfer cells, the aleurone layer, the endosperm, and the embryo. Cells from these tissues were isolated with the 'laser capture microdissection' technology and the extracted RNA was subjected to three rounds of T7-based amplification. The amplified RNA was subsequently hybridized to Affymetrix 22K Barley GeneChips. Due to the short average length of the amplified transcripts and the positioning of numerous probe sets at locations more than 400 base pairs (bp) from the poly(A)-tail, a normalization approach was used where the probe positions were taken into account. On the basis of the expression levels of a number of metal homeostasis genes, a working model is proposed for the translocation of zinc from the phloem to the storage sites in the developing grain. PMID- 19297553 TI - The patient-centered medical home and nephrology. PMID- 19297554 TI - C/EBP-beta modulates transcription of tubulointerstitial nephritis antigen in obstructive uropathy. AB - Tubulointerstitial injury leading to fibrosis is a common pathway of many renal diseases. During this type of injury, modeled by unilateral ureteral obstruction (UUO), cells undergo epithelial-to-mesenchymal transition (EMT), a process that is mediated by various cytokines that modulate the biology of extracellular matrix proteins. Here, we studied the tubulointerstitial nephritis antigen (TINag), a tubular basement membrane protein, in the UUO model of tubulointerstitial injury. We observed upregulation of type IV collagen but downregulation of both laminin and TINag in obstructed kidneys. TINag downregulation was a result of oxidative stress; in the proximal tubular epithelial cell line HK-2, TINag expression and its promoter activity decreased after treatment with H2O2. We identified multiple CCAAT/enhancer binding protein beta (C/EBP-beta) motifs in the TINag promoter and observed that oxidant stress perturbed interactions between TINag DNA and C/EBP-beta protein. Oxidant stress reduced nuclear translocation of C/EBP-beta in HK-2 cells, which was restored by antioxidants. In addition, overexpression of C/EBP-beta restored the H2O2-induced reduction of TINag promoter activity and expression. Furthermore, in vivo, renal obstruction reduced nuclear expression of C/EBP-beta. Cells grown on a TINag substratum maintained their normal epithelial phenotype and cytoskeletal organization, similar to those grown on type IV collagen, and demonstrated reduced synthesis of fibronectin. Taken together, these findings suggest that altered interactions between C/EBP-beta and TINag play a critical role in the pathophysiology of renal injury after obstruction. PMID- 19297555 TI - Anticoagulant and antiplatelet usage associates with mortality among hemodialysis patients. AB - Many prescribe anticoagulants and antiplatelet medications to prevent thromboembolic events and access thrombosis in dialysis patients despite limited evidence of their efficacy in this population. This retrospective cohort study examined whether use of warfarin, clopidogrel, and/or aspirin affected survival in 41,425 incident hemodialysis patients during 5 yr of follow-up. The prescription frequencies for warfarin, clopidogrel, and aspirin were 8.3, 10.0, and 30.4%, respectively, during the first 90 d of initiating chronic hemodialysis. Compared with the 24,740 patients receiving none of these medications, Cox proportional hazards analysis suggested that exposure to these medications was associated with increased risk for mortality (warfarin hazard ratio [HR] 1.27 [95% confidence interval (CI) 1.18 to 1.37]; clopidogrel HR 1.24 [95% CI 1.13 to 1.35]; and aspirin HR 1.06 [95% CI 1.01 to 1.11]). The increased mortality associated with warfarin or clopidogrel use remained in stratified analyses. A covariate- and propensity-adjusted time-varying analysis, which accounted for longitudinal changes in prescription, produced similar results. In addition, matching for treatment facility and attending physician revealed similar associations between prescription and mortality. We conclude that warfarin, aspirin, or clopidogrel prescription is associated with higher mortality among hemodialysis patients. Given the possibility of confounding by indication, randomized trials are needed to determine definitively the risk and benefit of these medications. PMID- 19297556 TI - Randomized, controlled trial of prednisone, cyclophosphamide, and cyclosporine in lupus membranous nephropathy. AB - Patients with lupus membranous nephropathy (LMN) are at substantial long-term risk for morbidity and mortality associated with protracted nephrotic syndrome, including ESRD. The optimal treatment for this condition is controversial. Forty two patients with LMN participated in a randomized, controlled trial to compare adjunctive immunosuppressive drugs with prednisone alone. Adjunctive regimens included either cyclosporine (CsA) for 11 mo or alternate-month intravenous pulse cyclophosphamide (IVCY) for six doses; the control group received alternate-day prednisone alone. Median proteinuria was 5.4 g/d (range 2.7 to 15.4 g/d). We assessed the primary outcome, time to remission of proteinuria during the 12-mo protocol, by univariate survival analysis. At 1 yr, the cumulative probability of remission was 27% with prednisone, 60% with IVCY, and 83% with CsA. Although both IVCY and CsA were more effective than prednisone in inducing remissions of proteinuria, relapse of nephrotic syndrome occurred significantly more often after completion of CsA than after IVCY. By multivariate survival analysis, treatment with prednisone and high-grade proteinuria (>5 g/d) but not race or ethnicity were independently associated with a decreased probability of remission. Adverse effects during the 12-mo protocol included insulin-requiring diabetes (one with prednisone and two with CsA), pneumonia (one with prednisone and two with CsA), and localized herpes zoster (two with IVCY). In conclusion, regimens containing CsA or IVCY are each more effective than prednisone alone in inducing remission of proteinuria among patients with LMN. PMID- 19297557 TI - Wnt/beta-catenin signaling promotes renal interstitial fibrosis. AB - Wnts compose a family of signaling proteins that play an essential role in kidney development, but their expression in adult kidney is thought to be silenced. Here, we analyzed the expression and regulation of Wnts and their receptors and antagonists in normal and fibrotic kidneys after obstructive injury. In the normal mouse kidney, the vast majority of 19 different Wnts and 10 frizzled receptor genes was expressed at various levels. After unilateral ureteral obstruction, all members of the Wnt family except Wnt5b, Wnt8b, and Wnt9b were upregulated in the fibrotic kidney with distinct dynamics. In addition, the expression of most Fzd receptors and Wnt antagonists was also induced. Obstructive injury led to a dramatic accumulation of beta-catenin in the cytoplasm and nuclei of renal tubular epithelial cells, indicating activation of the canonical pathway of Wnt signaling. Numerous Wnt/beta-catenin target genes (c Myc, Twist, lymphoid enhancer-binding factor 1, and fibronectin) were induced, and their expression was closely correlated with renal beta-catenin abundance. Delivery of the Wnt antagonist Dickkopf-1 gene significantly reduced renal beta catenin accumulation and inhibited the expression of Wnt/beta-catenin target genes. Furthermore, gene therapy with Dickkopf-1 inhibited myofibroblast activation; suppressed expression of fibroblast-specific protein 1, type I collagen, and fibronectin; and reduced total collagen content in the model of obstructive nephropathy. In summary, these results establish a role for Wnt/beta catenin signaling in the pathogenesis of renal fibrosis and identify this pathway as a potential therapeutic target. PMID- 19297558 TI - Placental insufficiency associated with loss of Cited1 causes renal medullary dysplasia. AB - A number of studies have shown that placental insufficiency affects embryonic patterning of the kidney and leads to a decreased number of functioning nephrons in adulthood; however, there is circumstantial evidence that placental insufficiency may also affect renal medullary growth, which could account for cases of unexplained renal medullary dysplasia and for abnormalities in renal function among infants who had experienced intrauterine growth retardation. We observed that mice with late gestational placental insufficiency associated with genetic loss of Cited1 expression in the placenta had renal medullary dysplasia. This was not caused by lower urinary tract obstruction or by defects in branching of the ureteric bud during early nephrogenesis but was associated with decreased tissue oxygenation and increased apoptosis in the expanding renal medulla. Loss of placental Cited1 was required for Cited1 mutants to develop renal dysplasia, and this was not dependent on alterations in embryonic Cited1 expression. Taken together, these findings suggest that renal medullary dysplasia in Cited1 mutant mice is a direct consequence of decreased tissue oxygenation resulting from placental insufficiency. PMID- 19297559 TI - Kidney-derived stromal cells modulate dendritic and T cell responses. AB - Multipotent mesenchymal stromal cells from the bone marrow ameliorate acute kidney injury through a mechanism other than transdifferentiation into renal tissue. Stromal cells exert immunoregulatory effects on dendritic and T cells, both of which are important in the pathophysiology of immune-mediated kidney injury. We hypothesized that similar cells with immunoregulatory function exist within the adult kidney. We isolated murine kidney-derived cells with morphologic features, growth properties, and an immunophenotype characteristic of mesenchymal stromal cells. These cells lacked lineage markers and could be differentiated into mesodermal cell lineages, including osteocytes and adipocytes. Furthermore, these kidney-derived cells induced the generation of bone marrow-derived dendritic cells with significantly reduced MHC II expression, increased CD80 expression, increased IL-10 production and the inability to stimulate CD4+ T cell proliferation in allogeneic and nominal antigen-specific cultures. Experiments in mixed and transwell cultures demonstrated that the production of soluble immune modulators, such as IL-6, was responsible for these effects on dendritic cell differentiation and maturation. Contact-dependent mechanisms, however, inhibited mitogenic T cell proliferation. In summary, kidney-derived cells may suppress inflammation in the kidney in vivo; a better understanding of their biology could have therapeutic implications in a wide variety of immune-mediated kidney diseases. PMID- 19297560 TI - Regulatory allospecific T cell clones abrogate chronic allograft rejection. AB - True alloantigen-specific tolerance is the ultimate goal of solid organ transplantation, eliminating the need for long-term immunosuppression. Recent evidence suggests that Th1-derived cytokines are associated with rejection and Th2-derived cytokines with long-term allograft survival, but the roles of these subsets in rejection and tolerance are incompletely understood. Here, we analyzed the functional and regulatory capacities of T cell clones derived from tolerant and rejecting rats (Wistar rat donors, Lewis rat recipients). We generated and subcloned T cell lines from lymphocytes derived from either acutely rejecting grafts or from the grafts of CTLA4-Ig-treated tolerant rats. Pretransplantation adoptive transfer of T cell clones generated from rejected grafts (Th1 clones) accelerated acute rejection or promoted development of chronic rejection, whereas transfer of T cell clones generated from tolerized grafts (Th2 clones) protected rats from acute rejection and progressive organ dysfunction. When Th1 and Th2 clones were injected simultaneously, Th2 clones specifically regulated activation of Th1 clones. Rats that received injections of Th2 clones accepted long-term donor-specific skin grafts but acutely rejected third-party skin grafts. Tolerant rats treated with Th2 clones demonstrated an increased number of regulatory CD4+CD25+Foxp3+ cells and strong mononuclear cell staining for IL-10 but negligible IFN-gamma, IL-17, and IL-23 compared with untreated rats or those treated with Th1 clones. In summary, these results demonstrate the regulatory functions of Th2 cells in a clinically relevant allogeneic transplant model and provide new insight into the functional role of Th2 cells in preventing the process of chronic rejection. PMID- 19297562 TI - The Medical Device Safety Act of 2009. PMID- 19297563 TI - Screening for prostate cancer. PMID- 19297561 TI - Enhanced interferon signaling pathway in oral cancer revealed by quantitative proteome analysis of microdissected specimens using 16O/18O labeling and integrated two-dimensional LC-ESI-MALDI tandem MS. AB - Oral squamous cell carcinoma (OSCC) remains one of the most common cancers worldwide, and the mortality rate of this disease has increased in recent years. No molecular markers are available to assist with the early detection and therapeutic evaluation of OSCC; thus, identification of differentially expressed proteins may assist with the detection of potential disease markers and shed light on the molecular mechanisms of OSCC pathogenesis. We performed a multidimensional (16)O/(18)O proteomics analysis using an integrated ESI-ion trap and MALDI-TOF/TOF MS system and a computational data analysis pipeline to identify proteins that are differentially expressed in microdissected OSCC tumor cells relative to adjacent non-tumor epithelia. We identified 1233 unique proteins in microdissected oral squamous epithelia obtained from three pairs of OSCC specimens with a false discovery rate of <3%. Among these, 977 proteins were quantified between tumor and non-tumor cells. Our data revealed 80 dysregulated proteins (53 up-regulated and 27 down-regulated) when a 2.5-fold change was used as the threshold. Immunohistochemical staining and Western blot analyses were performed to confirm the overexpression of 12 up-regulated proteins in OSCC tissues. When the biological roles of 80 differentially expressed proteins were assessed via MetaCore analysis, the interferon (IFN) signaling pathway emerged as one of the most significantly altered pathways in OSCC. As many as 20% (10 of 53) of the up-regulated proteins belonged to the IFN-stimulated gene (ISG) family, including ubiquitin cross-reactive protein (UCRP)/ISG15. Using head-and-neck cancer tissue microarrays, we determined that UCRP is overexpressed in the majority of cheek and tongue cancers and in several cases of larynx cancer. In addition, we found that IFN-beta stimulates UCRP expression in oral cancer cells and enhances their motility in vitro. Our findings shed new light on OSCC pathogenesis and provide a basis for the future development of novel biomarkers. PMID- 19297564 TI - Screening for prostate cancer--the controversy that refuses to die. PMID- 19297567 TI - Images in clinical medicine. "Popcorn" calcifications in a pulmonary chondroid hamartoma. PMID- 19297568 TI - An uncertain future for cardiovascular drug development? PMID- 19297569 TI - Eliminating racial and ethnic disparities in cardiac care. PMID- 19297566 TI - Screening and prostate-cancer mortality in a randomized European study. AB - BACKGROUND: The European Randomized Study of Screening for Prostate Cancer was initiated in the early 1990s to evaluate the effect of screening with prostate specific-antigen (PSA) testing on death rates from prostate cancer. METHODS: We identified 182,000 men between the ages of 50 and 74 years through registries in seven European countries for inclusion in our study. The men were randomly assigned to a group that was offered PSA screening at an average of once every 4 years or to a control group that did not receive such screening. The predefined core age group for this study included 162,243 men between the ages of 55 and 69 years. The primary outcome was the rate of death from prostate cancer. Mortality follow-up was identical for the two study groups and ended on December 31, 2006. RESULTS: In the screening group, 82% of men accepted at least one offer of screening. During a median follow-up of 9 years, the cumulative incidence of prostate cancer was 8.2% in the screening group and 4.8% in the control group. The rate ratio for death from prostate cancer in the screening group, as compared with the control group, was 0.80 (95% confidence interval [CI], 0.65 to 0.98; adjusted P=0.04). The absolute risk difference was 0.71 death per 1000 men. This means that 1410 men would need to be screened and 48 additional cases of prostate cancer would need to be treated to prevent one death from prostate cancer. The analysis of men who were actually screened during the first round (excluding subjects with noncompliance) provided a rate ratio for death from prostate cancer of 0.73 (95% CI, 0.56 to 0.90). CONCLUSIONS: PSA-based screening reduced the rate of death from prostate cancer by 20% but was associated with a high risk of overdiagnosis. (Current Controlled Trials number, ISRCTN49127736.) PMID- 19297570 TI - Denial. PMID- 19297571 TI - Racial differences in incident heart failure among young adults. AB - BACKGROUND: The antecedents and epidemiology of heart failure in young adults are poorly understood. METHODS: We prospectively assessed the incidence of heart failure over a 20-year period among 5115 blacks and whites of both sexes who were 18 to 30 years of age at baseline. Using Cox models, we examined predictors of hospitalization or death from heart failure. RESULTS: Over the course of 20 years, heart failure developed in 27 participants (mean [+/-SD] age at onset, 39+/-6 years), all but 1 of whom were black. The cumulative incidence of heart failure before the age of 50 years was 1.1% (95% confidence interval [CI], 0.6 to 1.7) in black women, 0.9% (95% CI, 0.5 to 1.4) in black men, 0.08% (95% CI, 0.0 to 0.5) in white women, and 0% (95% CI, 0 to 0.4) in white men (P=0.001 for the comparison of black participants and white participants). Among blacks, independent predictors at 18 to 30 years of age of heart failure occurring 15 years, on average, later included higher diastolic blood pressure (hazard ratio per 10.0 mm Hg, 2.1; 95% CI, 1.4 to 3.1), higher body-mass index (the weight in kilograms divided by the square of the height in meters) (hazard ratio per 5.7 units, 1.4; 95% CI, 1.0 to 1.9), lower high-density lipoprotein cholesterol (hazard ratio per 13.3 mg per deciliter [0.34 mmol per liter], 0.6; 95% CI, 0.4 to 1.0), and kidney disease (hazard ratio, 19.8; 95% CI, 4.5 to 87.2). Three quarters of those in whom heart failure subsequently developed had hypertension by the time they were 40 years of age. Depressed systolic function, as assessed on a study echocardiogram when the participants were 23 to 35 years of age, was independently associated with the development of heart failure 10 years, on average, later (hazard ratio for abnormal systolic function, 36.9; 95% CI, 6.9 to 198.3; hazard ratio for borderline systolic function, 3.5; 95% CI, 1.2 to 10.2). Myocardial infarction, drug use, and alcohol use were not associated with the risk of heart failure. CONCLUSIONS: Incident heart failure before 50 years of age is substantially more common among blacks than among whites. Hypertension, obesity, and systolic dysfunction that are present before a person is 35 years of age are important antecedents that may be targets for the prevention of heart failure. (ClinicalTrials.gov number, NCT00005130.) PMID- 19297572 TI - Vaccine prevention of maternal cytomegalovirus infection. AB - BACKGROUND: Congenital infection with cytomegalovirus (CMV) is an important cause of hearing, cognitive, and motor impairments in newborns. METHODS: In this phase 2, placebo-controlled, randomized, double-blind trial, we evaluated a vaccine consisting of recombinant CMV envelope glycoprotein B with MF59 adjuvant, as compared with placebo. Three doses of the CMV vaccine or placebo were given at 0, 1, and 6 months to CMV-seronegative women within 1 year after they had given birth. We tested for CMV infection in the women in quarterly tests during a 42 month period, using an assay for IgG antibodies against CMV proteins other than glycoprotein B. Infection was confirmed by virus culture or immunoblotting. The primary end point was the time until the detection of CMV infection. RESULTS: We randomly assigned 234 subjects to receive the CMV vaccine and 230 subjects to receive placebo. A scheduled interim analysis led to a stopping recommendation because of vaccine efficacy. After a minimum of 1 year of follow-up, there were 49 confirmed infections, 18 in the vaccine group and 31 in the placebo group. Kaplan-Meier analysis showed that the vaccine group was more likely to remain uninfected during a 42-month period than the placebo group (P=0.02). Vaccine efficacy was 50% (95% confidence interval, 7 to 73) on the basis of infection rates per 100 person-years. One congenital infection among infants of the subjects occurred in the vaccine group, and three infections occurred in the placebo group. There were more local reactions (pain, erythema, induration, and warmth) and systemic reactions (chills, arthralgias, and myalgias) in the vaccine group than in the placebo group. CONCLUSIONS: CMV glycoprotein B vaccine has the potential to decrease incident cases of maternal and congenital CMV infection. (ClinicalTrials.gov number, NCT00125502.) PMID- 19297573 TI - Genetic compensation in a human genomic disorder. AB - Cytogenetic studies of the parents of a girl with the DiGeorge (or velocardiofacial) syndrome, who carried a deletion at 22q11.2, revealed an unexpected rearrangement of both 22q11.2 regions in the unaffected father. He carried a 22q11.2 deletion on one copy of chromosome 22 and a reciprocal 22q11.2 duplication on the other copy of chromosome 22. Genetic compensation, which is consistent with the normal phenotype of the father, was shown through quantitative-expression analyses of genes located within the genetic region associated with the DiGeorge syndrome. This finding has implications for genetic counseling and represents a case of genetic compensation in a human genomic disorder. PMID- 19297574 TI - Clinical practice. Carbon monoxide poisoning. PMID- 19297575 TI - Moyamoya disease and moyamoya syndrome. PMID- 19297576 TI - Images in clinical medicine. Pelvic-organ prolapse and uterine inversion. PMID- 19297577 TI - Case records of the Massachusetts General Hospital. Case 9-2009. An 81-year-old man with massive rectal bleeding. PMID- 19297565 TI - Mortality results from a randomized prostate-cancer screening trial. AB - BACKGROUND: The effect of screening with prostate-specific-antigen (PSA) testing and digital rectal examination on the rate of death from prostate cancer is unknown. This is the first report from the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial on prostate-cancer mortality. METHODS: From 1993 through 2001, we randomly assigned 76,693 men at 10 U.S. study centers to receive either annual screening (38,343 subjects) or usual care as the control (38,350 subjects). Men in the screening group were offered annual PSA testing for 6 years and digital rectal examination for 4 years. The subjects and health care providers received the results and decided on the type of follow-up evaluation. Usual care sometimes included screening, as some organizations have recommended. The numbers of all cancers and deaths and causes of death were ascertained. RESULTS: In the screening group, rates of compliance were 85% for PSA testing and 86% for digital rectal examination. Rates of screening in the control group increased from 40% in the first year to 52% in the sixth year for PSA testing and ranged from 41 to 46% for digital rectal examination. After 7 years of follow-up, the incidence of prostate cancer per 10,000 person-years was 116 (2820 cancers) in the screening group and 95 (2322 cancers) in the control group (rate ratio, 1.22; 95% confidence interval [CI], 1.16 to 1.29). The incidence of death per 10,000 person-years was 2.0 (50 deaths) in the screening group and 1.7 (44 deaths) in the control group (rate ratio, 1.13; 95% CI, 0.75 to 1.70). The data at 10 years were 67% complete and consistent with these overall findings. CONCLUSIONS: After 7 to 10 years of follow-up, the rate of death from prostate cancer was very low and did not differ significantly between the two study groups. (ClinicalTrials.gov number, NCT00002540.) PMID- 19297578 TI - One step closer to a CMV vaccine. PMID- 19297579 TI - RTS,S/AS01E vaccine against malaria. PMID- 19297580 TI - Antimalarial therapies in children from Papua New Guinea. PMID- 19297581 TI - Smoking exposure, 17q21 variants, and early-onset asthma. PMID- 19297582 TI - Irbesartan for heart failure with preserved ejection fraction. PMID- 19297583 TI - Medical mystery: Constipation--the answer. PMID- 19297584 TI - Cytokine induction after laboratory-acquired West Nile virus infection. PMID- 19297585 TI - Rice virescent3 and stripe1 encoding the large and small subunits of ribonucleotide reductase are required for chloroplast biogenesis during early leaf development. AB - The virescent3 (v3) and stripe1 (st1) mutants in rice (Oryza sativa) produce chlorotic leaves in a growth stage-dependent manner under field conditions. They are temperature-conditional mutants that produce bleached leaves at a constant 20 degrees C or 30 degrees C but almost green leaves under diurnal 30 degrees C/20 degrees C conditions. Here, we show V3 and St1, which encode the large and small subunits of ribonucleotide reductase (RNR), RNRL1, and RNRS1, respectively. RNR regulates the rate of deoxyribonucleotide production for DNA synthesis and repair. RNRL1 and RNRS1 are highly expressed in the shoot base and in young leaves, and the expression of the genes that function in plastid transcription/translation and in photosynthesis is altered in v3 and st1 mutants, indicating that a threshold activity of RNR is required for chloroplast biogenesis in developing leaves. There are additional RNR homologs in rice, RNRL2 and RNRS2, and eukaryotic RNRs comprise alpha(2)beta(2) heterodimers. In yeast, RNRL1 interacts with RNRS1 (RNRL1:RNRS1) and RNRL2:RNRS2, but no interaction occurs between other combinations of the large and small subunits. The interacting activities are RNRL1:RNRS1 > RNRL1:rnrs1(st1) > rnrl1(v3):RNRS1 > rnrl1(v3):rnrs1(st1), which correlate with the degree of chlorosis for each genotype. This suggests that missense mutations in rnrl1(v3) and rnrs1(st1) attenuate the first alphabeta dimerization. Moreover, wild-type plants exposed to a low concentration of an RNR inhibitor, hydroxyurea, produce chlorotic leaves without growth retardation, reminiscent of v3 and st1 mutants. We thus propose that upon insufficient activity of RNR, plastid DNA synthesis is preferentially arrested to allow nuclear genome replication in developing leaves, leading to continuous plant growth. PMID- 19297586 TI - An aldehyde oxidase in developing seeds of Arabidopsis converts benzaldehyde to benzoic Acid. AB - Arabidopsis (Arabidopsis thaliana) siliques synthesize high levels of benzoic acid (BA), which is incorporated into several glucosinolate compounds. The origin of BA in the siliques has not yet been determined. Here, we show that siliques have higher levels of benzaldehyde (BD)-oxidizing activity relative to leaves. The BD-oxidizing activity was purified from siliques in several chromatographic steps, and a 145-kD protein was identified as the enzyme most likely to possess this activity. The protein was trypsinized, and the sequence of the resulting peptides was determined by mass spectrometry, identifying it as the product of gene At1g04580, also designated as AAO4 (for ARABIDOPSIS ALDEHYDE OXIDASE4). AAO4 had previously been shown to be highly and specifically expressed in developing seeds, and its protein was shown to belong to a family of aldehyde oxidases. Here, we show that the AAO4 protein is an aldehyde oxidase that can use several substrates but that, among the substrates tested, has the lowest K(m) value (23 microm) with BD. AAO4 is able to oxidize BD without NAD(+), but its activity increases by 50% when this cofactor is added. The pH optimum of AAO4 is 7.0. Plants homozygous for a null allele in AAO4 showed a reduction of 30% to 45% in the total levels of BA in seeds as well as 7% to 9% and 32% to 38% decreases in the levels of 3-benzoyloxypropylglucosinolate and 4-benzoyloxybutylglucosinolate, respectively. Expressing AAO4 in Escherichia coli resulted in a 3-fold increase of BD-oxidizing activity in crude bacterial extracts over endogenous levels. These findings indicate that in Arabidopsis seeds, oxidation of BD contributes in part to the synthesis of BA. PMID- 19297587 TI - Grapevine MATE-type proteins act as vacuolar H+-dependent acylated anthocyanin transporters. AB - In grapevine (Vitis vinifera), anthocyanins are responsible for most of the red, blue, and purple pigmentation found in the skin of berries. In cells, anthocyanins are synthesized in the cytoplasm and accumulated into the vacuole. However, little is known about the transport of these compounds through the tonoplast. Recently, the sequencing of the grapevine genome allowed us to identify genes encoding proteins with high sequence similarity to the Multidrug And Toxic Extrusion (MATE) family. Among them, we selected two genes as anthocyanin transporter candidates and named them anthoMATE1 (AM1) and AM3. The expression of both genes was mainly fruit specific and concomitant with the accumulation of anthocyanin pigment. Subcellular localization assays in grapevine hairy roots stably transformed with AM1 or AM3green fluorescent protein fusion protein revealed that AM1 and AM3 are primarily localized to the tonoplast. Yeast vesicles expressing anthoMATEs transported acylated anthocyanins in the presence of MgATP. Inhibitor studies demonstrated that AM1 and AM3 proteins act in vitro as vacuolar H(+)-dependent acylated anthocyanin transporters. By contrast, under our experimental conditions, anthoMATEs could not transport malvidin 3-O glucoside or cyanidin 3-O-glucoside, suggesting that the acyl conjugation was essential for the uptake. Taken together, these results provide evidence that in vitro the two grapevine AM1 and AM3 proteins mediate specifically acylated anthocyanin transport. PMID- 19297588 TI - Developmental and hormonal regulation of gibberellin biosynthesis and catabolism in pea fruit. AB - In pea (Pisum sativum), normal fruit growth requires the presence of the seeds. The coordination of growth between the seed and ovary tissues involves phytohormones; however, the specific mechanisms remain speculative. This study further explores the roles of the gibberellin (GA) biosynthesis and catabolism genes during pollination and fruit development and in seed and auxin regulation of pericarp growth. Pollination and fertilization events not only increase pericarp PsGA3ox1 message levels (codes for GA 3-oxidase that converts GA(20) to bioactive GA(1)) but also reduce pericarp PsGA2ox1 mRNA levels (codes for GA 2 oxidase that mainly catabolizes GA(20) to GA(29)), suggesting a concerted regulation to increase levels of bioactive GA(1) following these events. 4 Chloroindole-3-acetic acid (4-Cl-IAA) was found to mimic the seeds in the stimulation of PsGA3ox1 and the repression of PsGA2ox1 mRNA levels as well as the stimulation of PsGA2ox2 mRNA levels (codes for GA 2-oxidase that mainly catabolizes GA(1) to GA(8)) in pericarp at 2 to 3 d after anthesis, while the other endogenous pea auxin, IAA, did not. This GA gene expression profile suggests that both seeds and 4-Cl-IAA can stimulate the production, as well as modulate the half-life, of bioactive GA(1), leading to initial fruit set and subsequent growth and development of the ovary. Consistent with these gene expression profiles, deseeded pericarps converted [(14)C]GA(12) to [(14)C]GA(1) only if treated with 4-Cl-IAA. These data further support the hypothesis that 4 Cl-IAA produced in the seeds is transported to the pericarp, where it differentially regulates the expression of pericarp GA biosynthesis and catabolism genes to modulate the level of bioactive GA(1) required for initial fruit set and growth. PMID- 19297589 TI - Pseudomonas aeruginosa acute prostatitis and urosepsis after sexual relations in a hot tub. AB - We report a case of a previously healthy 38-year-old male with acute prostatitis and concurrent Pseudomonas aeruginosa urosepsis. Pulsed-field gel electrophoresis analysis confirmed that the source of the organism was the patient's newly purchased hot tub, which was filled with water from a stream. PMID- 19297590 TI - Rapid determination of quinolone resistance in Acinetobacter spp. AB - In the treatment of serious bacterial infections, the rapid institution of appropriate antimicrobial chemotherapy may be lifesaving. Choosing the correct antibiotic or combination of antibiotics is becoming very important, as multidrug resistance is found in many pathogens. Using a collection of 75 well characterized multidrug-resistant (MDR) Acinetobacter sp. isolates, we show that PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) and base composition analysis of PCR amplification products can quickly and accurately identify quinolone resistance mediated by mutations in the quinolone resistance determining regions of gyrA and parC, two essential housekeeping genes. Single point mutations detected by PCR/ESI-MS in parC (found in 55/75 of the isolates) and in gyrA (found in 66/75 of the isolates) correlated with susceptibility testing and sequencing. By targeting resistance determinants that are encoded by genes with highly conserved DNA sequences (e.g., gyrA and parC), we demonstrate that PCR/ESI-MS can provide critical information for resistance determinant identification and can inform therapeutic decision making in the treatment of Acinetobacter sp. infections. PMID- 19297591 TI - Brain abscess caused by Tsukamurella tyrosinosolvens in an immunocompetent patient. AB - We describe a previously healthy patient with chronic otitis media complicated with cerebellar abscess caused by Tsukamurella tyrosinosolvens. The organism was identified based on conventional biochemical identification methods, PCR restriction fragment length polymorphism analysis of the hsp65 gene, and 16S rRNA gene sequencing. The patient was treated successfully with debridements and prolonged antibiotic therapy. PMID- 19297592 TI - Automation of the BD GeneOhm methicillin-resistant Staphylococcus aureus assay for high-throughput screening of nasal swab specimens. AB - This study demonstrated that an automated version of the BD-GeneOhm methicillin resistant Staphylococcus aureus (MRSA) assay (BD-MRSA), using achromopeptidase sample lysis and PCR setup performed on the Hamilton MICROLAB STAR(let) (Auto MRSA), gave results comparable to those obtained with BD-MRSA. The positive- and negative-result concordance rates and overall concordance of BD-MRSA and Auto MRSA were 98.2, 97.7, and 97.6%, respectively. Auto-MRSA required 60% less technical time than BD-MRSA, and the time required to obtain results was 50% of that required for BD-MRSA. Auto-MRSA provides a reliable, robust system for high volume clinical use. PMID- 19297594 TI - High sensitivity and specificity of acid-fast microscopy for diagnosis of pulmonary tuberculosis in an African population with a high prevalence of human immunodeficiency virus. AB - Laboratories in low-income countries report that acid-fast microscopy is insensitive and nonspecific. We demonstrate that for a Ugandan population with high prevalences of tuberculosis and human immunodeficiency virus infection, acid fast microscopy is highly sensitive (93.1%) and specific (100%) when performed by trained technologists in a carefully controlled manner using established techniques. PMID- 19297593 TI - Rapid molecular genotyping and clonal complex assignment of Staphylococcus aureus isolates by PCR coupled to electrospray ionization-mass spectrometry. AB - We describe a high-throughput assay using PCR coupled to electrospray ionization mass spectrometry (PCR/ESI-MS) to determine the genotypes of Staphylococcus aureus isolates. The primer sets used in the PCR/ESI-MS assay were designed to amplify the same genes analyzed in multilocus sequence typing (MLST). The method was used to identify the clonal complex and USA type of each isolate and is suitable for use in a clinical or public-health setting. The method was validated using a panel of diverse isolates from the Centers for Disease Control and Prevention that were previously characterized by MLST and pulsed-field gel electrophoresis (PFGE). Clinical isolates from two geographically distinct hospitals were characterized, and the clustering results were in agreement with those for repetitive-element PCR and PFGE. The PCR/ESI-MS method enables genotyping of over 180 samples of S. aureus per day in an automated fashion. PMID- 19297595 TI - Microbiological and epidemiological features of clinical respiratory isolates of Burkholderia gladioli. AB - Burkholderia gladioli, primarily known as a plant pathogen, is involved in human infections, especially in patients with cystic fibrosis (CF). In the present study, the first respiratory isolates recovered from 14 French patients with CF and 4 French patients without CF, identified by 16S rRNA gene analysis, were tested for growth on B. cepacia selective media, for identification by commercial systems, and for their antimicrobial susceptibilities, and were compared by pulsed-field gel electrophoresis (PFGE). Patients' data were collected. All 18 isolates grew on oxidation-fermentation-polymyxin B-bacitracin-lactose medium and Pseudomonas cepacia agar, but only 13 grew on Burkholderia cepacia selective agar. API 20NE strips did not differentiate B. gladioli from B. cepacia, whereas Vitek 2 GN cards correctly identified 15 isolates. All isolates were susceptible to piperacillin, imipenem, aminoglycosides, and ciprofloxacin and were far less resistant to ticarcillin than B. cepacia complex organisms. Fifteen PFGE types were observed among the 18 isolates, but shared types were not identified among epidemiologically related patients. The microbiological follow-up of CF patients showed that colonization was persistent in 3 of 13 documented cases; B. gladioli was isolated from posttransplantation cultures of blood from 1 patient. Among the patients without CF, B. gladioli was associated with intubation (three cases) or bronchiectasis (one case). In summary, the inclusion of B. gladioli in the databases of commercial identification systems should improve the diagnostic capabilities of those systems. In CF patients, this organism is more frequently involved in transient infections than in chronic infections, but it may be responsible for complications posttransplantation; patient-to-patient transmission has not been demonstrated to date. Lastly, B. gladioli appears to be naturally susceptible to aminoglycosides and ciprofloxacin, although resistant isolates may emerge in the course of chronic infections. PMID- 19297596 TI - Rapid Identification of Mycobacterium tuberculosis and nontuberculous mycobacteria by multiplex, real-time PCR. AB - The rapid identification of mycobacteria from culture is of primary importance for the administration of empirical antibiotic therapy and for the implementation of public health measures, yet there are few commercially available assays that can easily and accurately identify the mycobacteria in culture in a timely manner. Here we report on the development of a multiplex, real-time PCR assay that can identify 93% of the pathogenic mycobacteria in our laboratory in two parallel reactions. The mycobacteria identified by this assay include the Mycobacterium tuberculosis complex (MTC), the M. avium complex (MAC), the M. chelonae-M. abscessus group (MCAG), the M. fortuitum group (MFG), and M. mucogenicum. The primer targets included the 16S rRNA gene and the internal transcribed spacer. The assay was initially validated with a repository of reference strains and was subsequently tested with 314 clinical cultures identified by the AccuProbe assay or high-performance liquid chromatography. Of the 314 cultures tested, multiplex, real-time PCR produced congruent results for 99.8% of the 1,559 targets evaluated. The sensitivity and the specificity were each 99% or greater for MTC (n = 96), MAC (n = 97), MCAG (n = 68), and M. mucogenicum (n = 9) and 95% and 100%, respectively, for MFG (n = 19). We conclude that this multiplex, real-time PCR assay is a useful diagnostic tool for the rapid and accurate identification of MTC and clinically relevant nontuberculous mycobacteria. PMID- 19297597 TI - Variations in ceftazidime and amoxicillin-clavulanate susceptibilities within a clonal infection of Burkholderia pseudomallei. AB - A patient with a clonal infection of Burkholderia pseudomallei had subpopulations with ceftazidime and amoxicillin-clavulanate susceptibilities that differed among the clinical specimens. Resistance was associated with a novel Cys69Tyr substitution in the Ambler class A beta-lactamase. Susceptibility testing of multiple colony variants from different sites should be performed for patients with culture-confirmed melioidosis. PMID- 19297598 TI - New pyrenochaeta species causing keratitis. AB - We report a new fungus as an agent of fungal keratitis in a diabetic woman. The fungal etiology was established by classic microbiology and PCR following 3 months of antibacterial therapy. The morphological features of the isolate and sequence analysis of the internal transcribed spacer region indicate a new species of Pyrenochaeta (Coelomycetes). PMID- 19297599 TI - Impact of discrepancies between the Abbott realtime and cobas TaqMan assays for quantification of human immunodeficiency virus type 1 group M non-B subtypes. AB - Viral loads in 249 clinical samples from individual patients infected with human immunodeficiency virus type 1 non-B subtypes were determined with both the Abbott RealTime and Cobas TaqMan assays. The differences exceeded 0.5 log for about 20% of samples and 1 log for 3%, with higher values always from the Abbott assay in the latter cases. PMID- 19297600 TI - A common variant of staphylococcal cassette chromosome mec type IVa in isolates from Copenhagen, Denmark, is not detected by the BD GeneOhm methicillin-resistant Staphylococcus aureus assay. AB - Rapid tests for detection of methicillin-resistant Staphylococcus aureus (MRSA) carriage are important to limit the transmission of MRSA in the health care setting. We evaluated the performance of the BD GeneOhm MRSA real-time PCR assay using a diverse collection of MRSA isolates, mainly from Copenhagen, Denmark, but also including international isolates, e.g., USA100-1100. Pure cultures of 349 MRSA isolates representing variants of staphylococcal cassette chromosome mec (SCCmec) types I to V and 103 different staphylococcal protein A (spa) types were tested. In addition, 53 methicillin-susceptible Staphylococcus aureus isolates were included as negative controls. Forty-four MRSA isolates were undetectable; of these, 95% harbored SCCmec type IVa, and these included the most-common clone in Copenhagen, spa t024-sequence type 8-IVa. The false-negative MRSA isolates were tested with new primers (analyte-specific reagent [ASR] BD GeneOhm MRSA assay) supplied by Becton Dickinson (BD). The ASR BD GeneOhm MRSA assay detected 42 of the 44 isolates that were false negative in the BD GeneOhm MRSA assay. Combining the BD GeneOhm MRSA assay with the ASR BD GeneOhm MRSA assay greatly improved the results, with only two MRSA isolates being false negative. The BD GeneOhm MRSA assay alone is not adequate for MRSA detection in Copenhagen, Denmark, as more than one-third of our MRSA isolates would not be detected. We recommend that the BD GeneOhm MRSA assay be evaluated against the local MRSA diversity before being established as a standard assay, and due to the constant evolution of SCCmec cassettes, a continuous global surveillance is advisable in order to update the assay as necessary. PMID- 19297601 TI - Changes in the agr locus affect enteritis caused by methicillin-resistant Staphylococcus aureus. AB - We studied the characteristics of methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) strains that caused enteritis. In a previous report, we demonstrated that both phenotypic and genotypic changes were associated with MRSA enteritis; and we hypothesized that the accessory gene regulator (agr), which is a global regulator of staphylococcal virulence and upregulates several exoproteins, is the key factor associated with the development of MRSA enteritis. In this study, we examined 12 MRSA isolates associated with enteritis from stool samples and 17 MRSA isolates not associated with enteritis that had the following characteristics: the strains associated with enteritis had the same genotype (genotype A), as detected by pulsed-field gel electrophoresis, or the strains were isolated from stools. The differences between strains that caused enteritis and those that did not cause enteritis strains were examined by quantitative reverse transcription-PCR to assess RNAII, agrA, RNAIII, and tst expression and by sequencing of the agr locus. The levels of expression of agrA, RNAIII, and tst were higher by the MRSA isolates associated with enteritis than by the MRSA isolates not associated with enteritis, whether or not they were of the same genotype. The levels of expression of RNAII by almost all the clinical isolates were similar. Sequencing of the agr locus showed that all MRSA isolates that caused enteritis have agr mutations, whereas the MRSA isolates that did not cause enteritis, with three exceptions, did not. Many of the isolates associated with enteritis had the same mutation, especially at the C-terminal end of agrA. These results suggest a trend in which mutations in the agr locus modify the expression of agrA and RNAIII and the production of toxin, all of which may increase the virulence and influence the occurrence of MRSA enteritis. PMID- 19297602 TI - Distribution of hepatitis B virus genotypes among patients with chronic infection in Japan shifting toward an increase of genotype A. AB - Acute hepatitis B virus (HBV) infection has been increasing through promiscuous sexual contacts, and HBV genotype A (HBV/A) is frequent in patients with acute hepatitis B (AHB) in Japan. To compare the geographic distribution of HBV genotypes in patients with chronic hepatitis B (CHB) in Japan between 2005 and 2006 and between 2000 and 2001, with special attention to changes in the proportion of HBV/A, a cohort study was performed to survey changes in genotypes of CHB patients at 16 hospitals throughout Japan. Furthermore, we investigated the clinical characteristics of each genotype and examined the genomic characteristics of HBV/A isolates by molecular evolutionary analyses. Of the 1,271 patients, 3.5%, 14.1%, and 82.3% were infected with HBV/A, -B, and -C, respectively. In comparison with our previous survey during 2000 and 2001, HBV/A was twice as frequent (3.5% versus 1.7%; P = 0.02). The mean age was lower in the patients with HBV/A than in those with HBV/B or -C. Based on phylogenetic analyses of 11 full-length genomes and 29 pre-S2/S region sequences from patients, HBV/A isolates were imported from Europe and the United States, as well as the Philippines and India. They clustered with HBV/A from AHB patients and have spread throughout Japan. HBV/A has been increasing in CHB patients in Japan as a consequence of AHB spreading in the younger generation through promiscuous sexual contacts, aided by a tendency of HBV/A to induce chronic hepatitis. The spread of HBV/A infection in Japan should be prevented by universal vaccination programs. PMID- 19297603 TI - Cholera outbreaks caused by an altered Vibrio cholerae O1 El Tor biotype strain producing classical cholera toxin B in Vietnam in 2007 to 2008. AB - Vibrio cholerae O1 isolates collected during cholera outbreaks occurring from late 2007 to early 2008 in northern Vietnam were revealed to represent an altered strain containing the RS1 element followed by a CTX prophage harboring El Tor type rstR and classical ctxB on the large chromosome. PMID- 19297604 TI - Potential of rapid diagnosis for controlling drug-susceptible and drug-resistant tuberculosis in communities where Mycobacterium tuberculosis infections are highly prevalent. AB - The long-term persistence of Mycobacterium tuberculosis in communities with high tuberculosis prevalence is a serious problem aggravated by the presence of drug resistant tuberculosis strains. Drug resistance in an individual patient is often discovered only after a long delay, particularly if the diagnosis is based on current culture-based drug sensitivity testing methods. During such delays, the patient may transmit tuberculosis to his or her contacts. Rapid diagnosis of drug resistance would be expected to reduce this transmission and hence to decrease the prevalence of drug-resistant strains. To investigate this quantitatively, a mathematical model was constructed, assuming a homogeneous population structure typical of communities in South Africa where tuberculosis incidence is high. Computer simulations performed with this model showed that current control strategies will not halt the spread of multidrug-resistant tuberculosis in such communities. The simulations showed that the rapid diagnosis of drug resistance can be expected to reduce the incidence of drug-resistant cases provided the additional measure of screening within the community is implemented. PMID- 19297605 TI - Location of antigenic sites recognized by monoclonal antibodies in the influenza A virus nucleoprotein molecule. AB - The locations of amino acid positions relevant to antigenic variation in the nucleoprotein (NP) of influenza virus are not conclusively known. We analysed the antigenic structure of influenza A virus NP by introducing site-specific mutations at amino acid positions presumed to be relevant for the differentiation of strain differences by anti-NP monoclonal antibodies. Mutant proteins were expressed in a prokaryotic system and analysed by performing ELISA with monoclonal antibodies. Four amino acid residues were found to determine four different antibody-binding sites. When mapped in a 3D X-ray model of NP, the four antigenically relevant amino acid positions were found to be located in separate physical sites of the NP molecule. PMID- 19297606 TI - Targeted delivery of hepatitis C virus-specific short hairpin RNA in mouse liver using Sendai virosomes. AB - Internal ribosome entry site (IRES)-mediated translation of input viral RNA is the initial required step for the replication of the positive-stranded genome of hepatitis C virus (HCV). We have shown previously the importance of the GCAC sequence near the initiator AUG within the stem and loop IV (SLIV) region in mediating ribosome assembly on HCV RNA. Here, we demonstrate selective inhibition of HCV-IRES-mediated translation using short hairpin (sh)RNA targeting the same site within the HCV IRES. sh-SLIV showed significant inhibition of viral RNA replication in a human hepatocellular carcinoma (Huh7) cell line harbouring a HCV monocistronic replicon. More importantly, co-transfection of infectious HCV-H77s RNA and sh-SLIV in Huh7.5 cells successfully demonstrated a significant decrease in viral RNA in HCV cell culture. Additionally, we report, for the first time, the targeted delivery of sh-SLIV RNA into mice liver using Sendai virosomes and demonstrate selective inhibition of HCV-IRES-mediated translation. Results provide the proof of concept that Sendai virosomes could be used for the efficient delivery of shRNAs into liver tissue to block HCV replication. PMID- 19297607 TI - Incorporation of influenza A virus genome segments does not absolutely require wild-type sequences. AB - The efficient incorporation of influenza virus genome segments into virions is mediated by cis-acting regions at both ends of the viral RNAs. It was shown previously that nt 16-26 at the 3' end of the non-structural (NS) viral RNA of influenza A virus are important for efficient virion incorporation and that nt 27 56 also contribute to this process. To understand further the signalling requirements for genome packaging, this study performed linker-scanning mutagenesis in the latter region and found that nt 27-35 made an appreciable contribution to the efficient incorporation of the NS segment. An NS vRNA library was then generated composed of an RNA population with randomized nucleotides at positions 16-35 such that the virus could select the sequences it required for virion incorporation. The sequences selected differed from the wild-type sequence and no conserved nucleotides were selected. The ability of non-wild-type sequences to function in this manner indicates that the incorporation of influenza A virus genome segments does not absolutely require specific sequences. PMID- 19297608 TI - RNA triphosphatase and guanylyl transferase activities are associated with the RNA polymerase protein L of rinderpest virus. AB - Rinderpest virus (RPV) large (L) protein is an integral part of the ribonucleoprotein (RNP) complex of the virus that is responsible for transcription and replication of the genome. Previously, we have shown that recombinant L protein coexpressed along with P protein (as the L-P complex) catalyses the synthesis of all viral mRNAs in vitro and the abundance of mRNAs follows a gradient of polarity, similar to the occurrence in vivo. In the present work, we demonstrate that the viral mRNAs synthesized in vitro by the recombinant L or purified RNP are capped and methylated at the N7 guanine position. RNP from the purified virions, as well as recombinant L protein, shows RNA triphosphatase (RTPase) and guanylyl transferase (GT) activities. L protein present in the RNP complex catalyses the removal of gamma-phosphate from triphosphate-ended 25 nt RNA generated in vitro representing the viral N-terminal mRNA 5' sequence. The L protein forms a covalent enzyme-guanylate intermediate with the GMP moiety of GTP, whose formation is inhibited by the addition of pyrophosphate; thus, it exhibits characteristics of cellular GTs. The covalent bond between the enzyme and nucleotide is acid labile and alkali stable, indicating the presence of phosphoamide linkage. The C-terminal region (aa 1717-2183) of RPV L protein alone exhibits the first step of GT activity needed to form a covalent complex with GMP, though it lacks the ability to transfer GMP to substrate RNA. Here, we describe the biochemical characterization of the newly found RTPase/GT activity of L protein. PMID- 19297609 TI - Differential expression of microRNAs in Marek's disease virus-transformed T lymphoma cell lines. AB - MicroRNAs (miRNAs) are increasingly recognized to play crucial roles in regulation of gene expression in different biological events, including many sporadic forms of cancer. However, despite the involvement of several viruses in inducing cancer, only a limited number of studies have been carried out to examine the miRNA expression signatures in virus-induced neoplasia, particularly in herpesvirus-induced tumours where virus-encoded miRNAs also contribute significantly to the miRNome of the tumour cell. Marek's disease (MD) is a naturally occurring, rapid-onset CD4+ T-cell lymphoma of poultry, induced by the highly contagious Marek's disease virus (MDV). High levels of expression of virus encoded miRNAs and altered expression of several host-encoded miRNAs were demonstrated in the MDV-transformed lymphoblastoid cell line MSB-1. In order to identify the miRNA expression signature specific to MDV-transformed cells, we examined the global miRNA expression profiles in seven distinct MDV-transformed cell lines by microarray analysis. This study revealed that, in addition to the high levels of MDV-encoded miRNAs, these MD tumour-derived lymphoblastoid cell lines showed altered expression of several host-encoded miRNAs. Comparison of the miRNA expression profiles of these cell lines with the MDV-negative, retrovirus transformed AVOL-1 cell line showed that miR-150 and miR-223 are downregulated irrespective of the viral aetiology, whereas downregulation of miR-155 was specific for MDV-transformed tumour cells. Thus, increased expression of MDV encoded miRNAs with specific downregulation of miR-155 can be considered as unique expression signatures for MD tumour cells. Analysis of the functional targets of these miRNAs would contribute to the understanding of the molecular pathways of MD oncogenicity. PMID- 19297610 TI - Phenotypic similarities and differences between UL37-deleted pseudorabies virus and herpes simplex virus type 1. AB - In the absence of the tegument protein pUL37, virion formation of pseudorabies virus (PrV) and herpes simplex virus type 1 (HSV-1) is severely impaired. Non enveloped nucleocapsids accumulate in clusters in the cytoplasm, whereas only a few enveloped particles can be detected. Although a contribution of pUL37 to nuclear egress of HSV-1 has been suggested, the nuclear stages of morphogenesis are not impaired in PrV-DeltaUL37-infected cells. Moreover, HSV-1 pUL37 has been described as essential for replication, whereas PrV is able to replicate productively without pUL37, although to lower titres than wild-type virus. Thus, there may be functional differences between the respective pUL37 proteins. This study compared the phenotypes of UL37-deleted PrV and HSV-1 in parallel assays, using a novel pUL37 deletion mutant of HSV-1 strain KOS, HSV-1DeltaUL37[86-1035]. Aggregates of seemingly 'naked' nucleocapsids were present in the cytoplasm of African green monkey (Vero) or rabbit kidney (RK13) cells infected with HSV 1DeltaUL37[86-1035] or PrV-DeltaUL37. Nuclear retention of nucleocapsids was not observed in either virus. However, in contrast to PrV-DeltaUL37, HSV 1DeltaUL37[86-1035] was unable to replicate productively in, and to form plaques on, either Vero or RK13 cells. Trans-complementation of respective deletion mutants with the heterologous pUL37 did not ensue. These data demonstrate that the conserved pUL37 in HSV-1 and PrV have similar but distinct functions. PMID- 19297611 TI - Downregulation of vaccinia virus intermediate and late promoters by host transcription factor YY1. AB - Approximately half of the intermediate and late gene transcriptional promoters of vaccinia virus have a binding site for the cellular transcription factor YY1 that overlaps the initiator elements. Depletion of YY1 using RNA interference enhanced the activity of these promoters, while overexpression of YY1 repressed their activity. Viral promoter nucleotide replacements that specifically impair the binding of YY1 mostly alleviated the transcriptional repression and correlated with the ability of YY1 to stably interact with the initiator DNAs in vitro. The transcriptional repression activity was localized to the C-terminal DNA-binding domain of the protein. These results indicate that YY1 functions to negatively regulate these vaccinia virus promoters by binding to their initiator elements. PMID- 19297612 TI - Association of proteinuria with race, cause of chronic kidney disease, and glomerular filtration rate in the chronic kidney disease in children study. AB - BACKGROUND AND OBJECTIVES: Proteinuria is associated with chronic kidney disease (CKD), and heavy proteinuria predicts a rapid decline in kidney function. However, the epidemiologic distribution of this important biomarker study is not well described in the pediatric CKD population. DESIGN, SETTING, PARTICIPANTS & MEASUREMENTS: This cross-sectional study of North American children with CKD examined the association of proteinuria among the baseline clinical variables in the cohort. Urinary protein-to-creatinine ratios (Up/c) were used to measure level of proteinuria. RESULTS: Of the 419 subjects studied, the median GFR as measured by iohexol disappearance (iGFR) was 42 ml/min per 1.73 m(2), median duration of CKD was six yr, and glomerular diseases accounted for 22% of the CKD diagnoses. Twenty-four percent of children had normal range (Up/c <0.2), 62% had significant, and 14% had nephrotic-range proteinuria (Up/c >2.0). A decrease in iGFR was associated with an increase in Up/c. At any level of GFR, a higher Up/c was associated with a glomerular cause of CKD and non-Caucasian race. Among subjects with a glomerular cause of CKD, Up/c was lower in subjects reporting utilization of renin-angiotensin system (RAS) antagonists (median Up/c = 0.93) compared with those who did not (median Up/c = 3.78). CONCLUSIONS: Proteinuria is associated with level of iGFR, cause of CKD, and race. The longitudinal study design of Chronic Kidney Disease in Children (CKiD) cohort study and the large number of subjects being studied has created an opportunity to better define the association between proteinuria and CKD progression. PMID- 19297613 TI - Increased myeloperoxidase activity and protein nitration are indicators of inflammation in patients with Chagas' disease. AB - In this study, we investigated whether inflammatory responses contribute to oxidative/nitrosative stress in patients with Chagas' disease. We used three tests (enzyme-linked immunosorbent assay, immuno-flow cytometry, and STAT-PAK immunochromatography) to screen human serum samples (n = 1,481) originating from Chiapas, Mexico, for Trypanosoma cruzi-specific antibodies. We identified 121 subjects who were seropositive for T. cruzi-specific antibodies, a finding indicative of an 8.5% seroprevalence in the rural population from Chiapas. Seropositive and seronegative subjects were examined for plasma levels of biomarkers of inflammation, i.e., myeloperoxidase (MPO), inducible nitric oxide synthase (iNOS), and xanthine oxidase (XOD), as well as for oxidative (advanced oxidation protein products [AOPPs]) and nitrosative (3-nitrotyrosine [3NT]) biomarkers. The seropositive subjects exhibited a significant increase in MPO activity and protein level, the indicator of neutrophil activation. Subsequently, a corresponding increase in AOPP contents, formed by MPO-dependent hypochlorous acid and chloramine formation, was noted in seropositive subjects. The plasma level of 3NT was significantly increased in seropositive subjects, yet we observed no change in XOD activity (O(2)(-) source) and nitrate/nitrite contents (denotes iNOS activation and NO production), which implied that direct peroxynitrite formation does not contribute to increased nitrosative damage in chagasic subjects. Instead, a positive correlation between increased MPO activity and protein 3NT formation was observed, which suggested to us that MPO-dependent formation of nitrylchloride that occurs in the presence of physiological NO and O(2)(-) concentrations contributes to protein nitration. Overall, our data demonstrate that T. cruzi-induced neutrophil activation is pathological and contributes to MPO-mediated collateral protein oxidative and nitrosative damage in human patients with Chagas' disease. Therapies capable of suppressing MPO activity may be useful in controlling the inflammation and oxidative/nitrosative pathology in chagasic cardiomyopathy. PMID- 19297614 TI - Development of a human-murine chimeric immunoglobulin M antibody for use in the serological detection of human flavivirus antibodies. AB - Current diagnosis of human flaviviral infections relies heavily on serological techniques such as the immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA). Broad application of this assay is hindered by a lack of standardized human positive-control sera that react with the wide variety of flaviviruses that can cause human disease, e.g., dengue virus (DENV), West Nile virus (WNV), yellow fever virus (YFV), Japanese encephalitis virus (JEV), and St. Louis encephalitis virus (SLEV). We have created a human-murine chimeric antibody combining the variable regions of the broadly flavivirus cross-reactive murine monoclonal antibody (MAb) 6B6C-1 and the constant region of human IgM to produce a standardized reagent capable of replacing human positive-control sera in a MAC-ELISA for the diagnosis of all human flaviviral infections. The human murine chimeric IgM antibody secreted from plasmid-transformed Sp2/0-Ag14 cells had a level of serological activity identical to that of 6B6C-1 as measured by ELISA, immunoblotting, and MAC-ELISA for multiple members of the flavivirus genus, including WNV, SLEV, YFV, DENV, and JEV. PMID- 19297615 TI - Potential use of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus expressed in Rachiplusia nu larvae as an immunogen for chickens. AB - The hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus (NDV) was obtained as a recombinant antigen in Rachiplusia nu larvae. When it was used as an immunogen in chickens, a solid immune response, including neutralizing antibodies, was detected, demonstrating the potential use of this simple and economic strategy in the design of recombinant anti-NDV vaccines. PMID- 19297616 TI - Structural arrangement of the transmission interface in the antigen ABC transport complex TAP. AB - The transporter associated with antigen processing (TAP) represents a focal point in the immune recognition of virally or malignantly transformed cells by translocating proteasomal degradation products into the endoplasmic reticulum lumen for loading of MHC class I molecules. Based on a number of experimental data and the homology to the bacterial ABC exporter Sav1866, we constructed a 3D structural model of the core TAP complex and used it to examine the interface between the transmembrane and nucleotide-binding domains (NBD) by cysteine scanning and cross-linking approaches. Herein, we demonstrate the functional importance of the newly identified X-loop in the NBD in coupling substrate binding to downstream events in the transport cycle. We further verified domain swapping in a heterodimeric ABC half-transporter complex by cysteine cross linking. Strikingly, either substrate binding or translocation can be blocked by cross-linking the X-loop to coupling helix 2 or 1, respectively. These results resolve the structural arrangement of the transmission interface and point to different functions of the cytosolic loops and coupling helices in substrate binding, signaling, and transport. PMID- 19297617 TI - Addition of a cholesterol group to an HIV-1 peptide fusion inhibitor dramatically increases its antiviral potency. AB - Peptides derived from the heptad repeat 2 (HR2) region of the HIV fusogenic protein gp41 are potent inhibitors of viral infection, and one of them, enfuvirtide, is used for the treatment of therapy-experienced AIDS patients. The mechanism of action of these peptides is binding to a critical intermediate along the virus-cell fusion pathway, and accordingly, increasing the affinity for the intermediate yields more potent inhibitors. We took a different approach, namely to increase the potency of the HR2 peptide inhibitor C34 by targeting it to the cell compartment where fusion occurs, and we show here that a simple, yet powerful way to accomplish this is attachment of a cholesterol group. C34 derivatized with cholesterol (C34-Chol) shows dramatically increased antiviral potency on a panel of primary isolates, with IC(90) values 15- to 300-fold lower than enfuvirtide and the second-generation inhibitor T1249, making C34-Chol the most potent HIV fusion inhibitor to date. Consistent with its anticipated mechanism of action, the antiviral activity of C34-Chol is unusually persistent: washing target cells after incubation with C34-Chol, but before triggering fusion, increases IC(50) only 7-fold, relative to a 400-fold increase observed for C34. Moreover, derivatization with cholesterol extends the half-life of the peptide in vivo. In the mouse, s.c. administration of 3.5 mg/kg C34-Chol yields a plasma concentration 24 h after injection >300-fold higher than the measured IC(90) values. Because the fusion machinery targeted by C34-Chol is similar in several other enveloped viruses, we believe that these findings may be of general utility. PMID- 19297618 TI - Inhibition of Myc-induced cell transformation by brain acid-soluble protein 1 (BASP1). AB - Cell transformation by the Myc oncoprotein involves transcriptional activation or suppression of specific target genes with intrinsic oncogenic or tumor suppressive potential, respectively. We have identified the BASP1 (CAP-23, NAP 22) gene as a novel target suppressed by Myc. The acidic 25-kDa BASP1 protein was originally isolated as a cortical cytoskeleton-associated protein from rat and chicken brain, but has also been found in other tissues and subcellular locations. BASP1 mRNA and protein expression is specifically suppressed in fibroblasts transformed by the v-myc oncogene, but not in cells transformed by other oncogenic agents. The BASP1 gene encompasses 2 exons separated by a 58-kbp intron and a Myc-responsive regulatory region at the 5' boundary of untranslated exon 1. Bicistronic expression of BASP1 and v-myc from a retroviral vector blocks v-myc-induced cell transformation. Furthermore, ectopic expression of BASP1 renders fibroblasts resistant to subsequent cell transformation by v-myc, and exogenous delivery of the BASP1 gene into v-myc-transformed cells leads to significant attenuation of the transformed phenotype. The inhibition of v-myc induced cell transformation by BASP1 also prevents the transcriptional activation or repression of known Myc target genes. Mutational analysis showed that the basic N-terminal domain containing a myristoylation site, a calmodulin binding domain, and a putative nuclear localization signal is essential for the inhibitory function of BASP1. Our results suggest that down-regulation of the BASP1 gene is a necessary event in myc-induced oncogenesis and define the BASP1 protein as a potential tumor suppressor. PMID- 19297619 TI - Malaria primes the innate immune response due to interferon-gamma induced enhancement of toll-like receptor expression and function. AB - Malaria-induced sepsis is associated with an intense proinflammatory cytokinemia for which the underlying mechanisms are poorly understood. It has been demonstrated that experimental infection of humans with Plasmodium falciparum primes Toll-like receptor (TLR)-mediated proinflammatory responses. Nevertheless, the relevance of this phenomenon during natural infection and, more importantly, the mechanisms by which malaria mediates TLR hyperresponsiveness are unclear. Here we show that TLR responses are boosted in febrile patients during natural infection with P. falciparum. Microarray analyses demonstrated that an extraordinary percentage of the up-regulated genes, including genes involving TLR signaling, had sites for IFN-inducible transcription factors. To further define the mechanism involved in malaria-mediated "priming," we infected mice with Plasmodium chabaudi. The human data were remarkably predictive of what we observed in the rodent malaria model. Malaria-induced priming of TLR responses correlated with increased expression of TLR mRNA in a TLR9-, MyD88-, and IFNgamma dependent manner. Acutely infected WT mice were highly susceptible to LPS-induced lethality while TLR9(-/-), IL12(-/-) and to a greater extent, IFNgamma(-/-) mice were protected. Our data provide unprecedented evidence that TLR9 and MyD88 are essential to initiate IL12 and IFNgamma responses and favor host hyperresponsiveness to TLR agonists resulting in overproduction of proinflammatory cytokines and the sepsis-like symptoms of acute malaria. PMID- 19297620 TI - Assembling stable hair cell tip link complex via multidentate interactions between harmonin and cadherin 23. AB - The hereditary hearing-vision loss disease Usher syndrome (USH) is caused by defects in several proteins, most of which form an integrated protein network called Usher interactome. Harmonin/Ush1C is a master scaffold in the assembly of the Usher protein complexes, because harmonin is known to bind to every protein in the Usher interactome. However, the biochemical and structural mechanism governing the Usher protein complex formation is largely unclear. Here, we report that the highly-conserved N-terminal fragment of harmonin (N-domain) immediately preceding its PDZ1 adopts an autonomously-folded domain. We discovered that the N domain specifically binds to a short internal peptide fragment of the cadherin 23 cytoplasmic domain. The structures of the harmonin N-domain alone and in complex with the cadherin 23 internal peptide fragment uncovered the detailed binding mechanism of this interaction between harmonin and cadherin 23. We further elucidated the harmonin PDZ domain-mediated cadherin 23 binding by solving the structure of the second harmonin PDZ domain in complex with the cadherin 23 carboxyl tail. The multidentate binding mode between harmonin and cadherin 23 provides a structural and biochemical basis for the harmonin-mediated assembly of stable tip link complex in the auditory hair cells. PMID- 19297621 TI - Ruling out and ruling in neural codes. AB - The subject of neural coding has generated much debate. A key issue is whether the nervous system uses coarse or fine coding. Each has different strengths and weaknesses and, therefore, different implications for how the brain computes. For example, the strength of coarse coding is that it is robust to fluctuations in spike arrival times; downstream neurons do not have to keep track of the details of the spike train. The weakness, though, is that individual cells cannot carry much information, so downstream neurons have to pool signals across cells and/or time to obtain enough information to represent the sensory world and guide behavior. In contrast, with fine coding, individual cells can carry much more information, but downstream neurons have to resolve spike train structure to obtain it. Here, we set up a strategy to determine which codes are viable, and we apply it to the retina as a model system. We recorded from all the retinal output cells an animal uses to solve a task, evaluated the cells' spike trains for as long as the animal evaluates them, and used optimal, i.e., Bayesian, decoding. This approach makes it possible to obtain an upper bound on the performance of codes and thus eliminate those that are insufficient, that is, those that cannot account for behavioral performance. Our results show that standard coarse coding (spike count coding) is insufficient; finer, more information-rich codes are necessary. PMID- 19297622 TI - Confinement effects on the kinetics and thermodynamics of protein dimerization. AB - In the cell, protein complexes form by relying on specific interactions between their monomers. Excluded volume effects due to molecular crowding would lead to correlations between molecules even without specific interactions. What is the interplay of these effects in the crowded cellular environment? We study dimerization of a model homodimer when the mondimers are free and when they are tethered to each other. We consider a structured environment: Two monomers first diffuse into a cavity of size L and then fold and bind within the cavity. The folding and binding are simulated by using molecular dynamics based on a simplified topology based model. The confinement in the cell is described by an effective molecular concentration C approximately L(-3). A two-state coupled folding and binding behavior is found. We show the maximal rate of dimerization occurred at an effective molecular concentration C(op) approximately = 1 mM, which is a relevant cellular concentration. In contrast, for tethered chains the rate keeps at a plateau when C < C(op) but then decreases sharply when C > C(op). For both the free and tethered cases, the simulated variation of the rate of dimerization and thermodynamic stability with effective molecular concentration agrees well with experimental observations. In addition, a theoretical argument for the effects of confinement on dimerization is also made. PMID- 19297623 TI - p110gamma and p110delta isoforms of phosphoinositide 3-kinase differentially regulate natural killer cell migration in health and disease. AB - The mechanisms that regulate NK cell trafficking are unclear. Phosphoinositide-3 kinases (PI3K) control cell motility and the p110gamma and p110delta isoforms are mostly expressed in leukocytes, where they transduce signals downstream of G protein coupled receptors (GPCR) or tyrosine kinase receptors, respectively. Here, we set out to determine the relative contribution of p110gamma and p110delta to NK cell migration in mice. Using a combination of single-cell imaging analysis of transgenic cells reporting on PI3K activity in real time and small molecule inhibitors of p110gamma and p110delta, we show here that the tyrosine-kinase coupled p110delta is linked to GPCR signaling and, depending on the GPCR, may even be preferentially activated over p110gamma. Using gene targeted mice, we showed that both isoforms were essential for NK cell chemotaxis to CXCL12 and to CCL3 and, in vivo, for normal NK cell migration during pregnancy and to the inflamed peritoneum. By contrast, only p110delta was indispensable for chemotaxis to S1P and CXCL10 and for NK cell distribution throughout lymphoid and nonlymphoid tissues and for extravasation to tumors. These results implicate p110delta downstream of GPCRs in NK cells and highlight its nonredundant role as a key regulator of NK cell trafficking in health and disease. PMID- 19297624 TI - Chloroplast ribonucleoprotein CP31A is required for editing and stability of specific chloroplast mRNAs. AB - Chloroplast ribonucleoproteins (cpRNPs) are nuclear-encoded, highly abundant, and light-regulated RNA binding proteins. They have been shown to be involved in chloroplast RNA processing and stabilization in vitro and are phylogenetically related to the well-described heterogeneous nuclear ribonucleoproteins (hnRNPs). cpRNPs have been found associated with mRNAs present in chloroplasts and have been regarded as nonspecific stabilizers of chloroplast transcripts. Here, we demonstrate that null mutants of the cpRNP family member CP31A exhibit highly specific and diverse defects in chloroplast RNA metabolism. First, analysis of cp31a and cp31a/cp31b double mutants uncovers that these 2 paralogous genes participate nonredundantly in a combinatorial fashion in processing a subset of chloroplast editing sites in vivo. Second, a genome-wide analysis of chloroplast transcript accumulation in cp31a mutants detected a virtually complete loss of the chloroplast ndhF mRNA and lesser reductions for specific other mRNAs. Fluorescence analyses show that the activity of the NADH dehydrogenase complex, which also includes the NdhF subunit, is defective in cp31a mutants. This indicates that cpRNPs are important in vivo for calibrating the expression levels of specific chloroplast mRNAs and impact chloroplast physiology. Taken together, the specificity and combinatorial aspects of cpRNP functions uncovered suggest that these chloroplast proteins are functional equivalents of nucleocytosolic hnRNPs. PMID- 19297625 TI - Endometrium as an early sensor of in vitro embryo manipulation technologies. AB - Implantation is crucial for placental development that will subsequently impact fetal growth and pregnancy success with consequences on postnatal health. We postulated that the pattern of genes expressed by the endometrium when the embryo becomes attached to the mother uterus could account for the final outcome of a pregnancy. As a model, we used the bovine species where the embryo becomes progressively and permanently attached to the endometrium from day 20 of gestation onwards. At that stage, we compared the endometrial genes profiles in the presence of an in vivo fertilized embryo (AI) with the endometrial patterns obtained in the presence of nuclear transfer (SCNT) or in vitro fertilized embryos (IVF), both displaying lower and different potentials for term development. Our data provide evidence that the endometrium can be considered as a biological sensor able to fine-tune its physiology in response to the presence of embryos whose development will become altered much later after the implantation process. Compared with AI, numerous biological functions and several canonical pathways with a major impact on metabolism and immune function were found to be significantly altered in the endometrium of SCNT pregnancies at implantation, whereas the differences were less pronounced with IVF embryos. Determining the limits of the endometrial plasticity at the onset of implantation should bring new insights on the contribution of the maternal environment to the development of an embryo and the success of pregnancy. PMID- 19297626 TI - Mathematical test models for superparametrization in anisotropic turbulence. AB - The complexity of anisotropic turbulent processes over a wide range of spatiotemporal scales in engineering turbulence and climate atmosphere ocean science requires novel computational strategies with the current and next generations of supercomputers. In these applications the smaller-scale fluctuations do not statistically equilibrate as assumed in traditional closure modeling and intermittently send significant energy to the large-scale fluctuations. Superparametrization is a novel class of seamless multi-scale algorithms that reduce computational labor by imposing an artificial scale gap between the energetic smaller-scale fluctuations and the large-scale fluctuations. The main result here is the systematic development of simple test models that are mathematically tractable yet capture key features of anisotropic turbulence in applications involving statistically intermittent fluctuations without local statistical equilibration, with moderate scale separation and significant impact on the large-scale dynamics. The properties of the simplest scalar test model are developed here and utilized to test the statistical performance of superparametrization algorithms with an imposed spectral gap in a system with an energetic -5/3 turbulent spectrum for the fluctuations. PMID- 19297627 TI - Truth without consequence: reality and recall in refugees fleeing persecution. PMID- 19297628 TI - Reducing inpatient suicide risk: using human factors analysis to improve observation practices. AB - In 1995, the Joint Commission began requiring that hospitals report reviewable sentinel events as a condition of maintaining accreditation. Since then, inpatient suicide has been the second most common sentinel event reported to the Joint Commission. The Joint Commission emphasizes the need for around-the-clock observation for inpatients assessed as at high risk for suicide. However, there is sparse literature on the observation of psychiatric patients and no systematic studies or recommendations for best practices. Medical errors can best be reduced by focusing on systems improvements rather than individual provider mistakes. The author describes how failure modes and effects analysis (FMEA) was used proactively by an inpatient psychiatric treatment team to improve psychiatric observation practices by identifying and correcting potential observation process failures. Collection and implementation of observation risk reduction strategies across health care systems is needed to identify best practices and to reduce inpatient suicides. PMID- 19297629 TI - Let's think about human factors, not human failings. AB - Doctors typically think about medical errors as potential causes of malpractice litigation, as failures by individuals, and as evidence of personal incompetence that may deserve sanctions. Other professions take a different view: designing of safer systems, rather than criticism and punishment, is the way to prevent unintentional mishaps. In his article, Jeffrey Janofsky shows how psychiatrists can think about making care systems safer for patients. He also provides a splendid example of how forensic psychiatrists should conceptualize legal and medical problems encountered in clinical practice. PMID- 19297630 TI - A guide to the forensic assessment of race-based traumatic stress reactions. AB - A large body of empirical evidence has accumulated over the past decade documenting the psychiatric and emotional consequences of racism and racial discrimination. Still, little has been written in the legal or psychiatric assessment literature that is focused on describing the direct and specific effects of racism and offering guidance to forensic psychiatrists in understanding, assessing, and treating the race-based stress reactions that may occur as a result of exposure to racial discrimination or harassment. This article uses the analysis of a case study to illustrate and extend previous work on the evaluation of racial discrimination by providing a guide to the forensic assessment of the psychiatric and emotional impact of race-based encounters--a guide that can be used both in preparing expert reports and in developing treatment approaches. PMID- 19297631 TI - The trauma of insidious racism. AB - Race-based traumatic stress assessments are difficult because of the insidious nature of racism, the lack of scientific research, and the disregard of the nondominant cultural experience. Although there is a large body of scientific literature about the harmful psychological effects of racism, most of that literature is not directly applicable to individual assessment. Carter and Forsyth begin to correct that deficiency. Implicit in their attempt is the beginnings of defining a race-based traumatic stress syndrome. At present, the psychiatric expert can be useful to the court through a psychological description of the evaluee within a racial-cultural context. PMID- 19297632 TI - Attention deficit hyperactivity disorder in prison: a treatment protocol. AB - Prescribing controlled substances in correctional settings can create challenges for security, nursing, and psychiatric staff. Some inmates, including those with functionally significant attention deficit hyperactivity disorder (ADHD), however, can benefit from such treatment. This article describes the development of a protocol for the treatment of prison inmates with ADHD that attempted to address a broad range of concerns including disparate diagnostic and treatment standards among prison psychiatrists, conflicts between stakeholders, and medication misuse and substance abuse among inmates. The protocol provides criteria in four main areas: diagnosis, current functional impairment, treatment in general, and treatment with stimulants. Stakeholders had mixed reactions to the protocol. PMID- 19297633 TI - The top ten reasons to limit prescription of controlled substances in prisons. AB - Implementation of a treatment protocol for prescribing stimulant medications in adult prisons where such a protocol and oversight did not previously exist is prudent management, clinically relevant, and appropriate. There are multiple challenges to using controlled substances in correctional settings, many of them a consequence of the very high prevalence of substance use disorders among inmates. This article describes 10 of the reasons to limit the prescription of controlled substances in prison, among them the implications for nursing and correctional security staff and the overwhelming of the already limited psychiatric resources available in most prisons. PMID- 19297634 TI - Potential side effects of androgen deprivation treatment in sex offenders. AB - Testosterone is an important hormone involved in sexual arousal, and, indeed, a profound reduction of testosterone levels may be helpful in controlling sexual impulses in sex offenders. Earlier thought of as a sex hormone only, testosterone has been increasingly shown to have manifold actions in the adult male. Normal adult levels of androgens are required for the health of bones, a large number of metabolic functions, mood, erythropoiesis, sebaceous gland activity of the skin, and several other functions. Severe androgen deficiency is associated with pathologies of these biological systems. Androgen deprivation therapy may result in osteoporosis, weight gain with an increased visceral adiposity, impaired glucose tolerance, dyslipidemia, and emotional disturbances. Some of these features combine in the metabolic syndrome that is also frequently associated with the use of psychotropic medication in general. It leads to a moderately increased risk of fractures and diabetes mellitus (by 40%-50%), and a small increased risk of cardiovascular morbidity and depression (by 10%-20%). It should be noted that small proportionate increases in risk may be of modest clinical significance when background risks are very low. Effective and safe management of sex offenders treated with testosterone-deprivation therapy should include careful monitoring of side effects and their prevention and treatment. PMID- 19297635 TI - Risk/benefit ratio of androgen deprivation treatment for sex offenders. AB - Androgen deprivation treatment (ADT) significantly lowers testosterone. That, in turn, can decrease sexual drive, facilitating better self-control and lower recidivism rates among sexually disordered offenders. Potential side effects can include: decreases in bone density; development of a metabolic syndrome involving weight gain, accompanied by changes in glucose and lipid metabolism; and rarely, depression. In the presence of a proper treatment protocol designed either to prevent or to minimize side effects, particularly the development of osteoporosis, the risks associated with ADT are generally within the same range as those associated with many other commonly prescribed psychotropic interventions. PMID- 19297636 TI - Expert witness confidence and juror personality: their impact on credibility and persuasion in the courtroom. AB - The present study was conducted to investigate the relationship between both expert witness confidence and juror personality with expert witness credibility, as well as expert witness credibility with juror sentencing outcome. Participants were presented with one of three randomly assigned filmed scenarios depicting various levels of manipulated witness confidence. They then completed a sentencing outcome item, the Witness Credibility Scale, and the Goldberg Five Factor Markers. Expert witness confidence had a significant main effect on ratings of credibility, with moderate levels of manipulated confidence yielding the highest credibility. Juror extroversion was positively related to perceptions of expert witness credibility. Finally, juror ratings of expert witness credibility, as well as two subcomponents, predicted juror sentencing outcome. PMID- 19297637 TI - Violence and substance use among female partners of men in treatment for intimate partner violence. AB - To improve understanding of the complex dynamics in intimate partner violence (IPV) in heterosexual relationships, we explored violence and substance use among the female partners of men entering treatment for both IPV and substance-related problems. All male participants (n = 75) were alcohol dependent and had at least one domestic-violence arrest. Results showed that female partners were as likely as men to engage in substance use the week before treatment; however, according to reports by the men, the female partners were more likely than men to use substances during the last week of treatment, due to a reported increase in use during the men's treatment. Regarding violence, 59 percent of female IPV victims reported engaging in some form of mild violence against their male partners, and 55 percent reported engaging in some form of severe violence. By contrast, only 23 percent of male batterers reported that their female partners had engaged in mild violence, and only 19 percent reported that their partners had engaged in severe violence. Regardless of whether the violence was defensive in nature, the data suggest that women in relationships involving substance abuse and IPV are in need of treatment. Implications of these findings are discussed. PMID- 19297638 TI - Suicide among incarcerated veterans. AB - Both veterans and jail/prison inmates face an increased risk of suicide. The incarcerated veteran sits at the intersection of these two groups, yet little is known about this subpopulation, particularly its risk of suicide. A Pubmed/Medline/PsycINFO search anchored to incarcerated veteran suicide, veteran suicide, suicide in jails/prisons, and veterans incarcerated from 2000 to the present was performed. The currently available literature does not reveal the suicide risk of incarcerated veterans, nor does it enable meaningful estimates. However, striking similarities and overlapping characteristics link the data on veteran suicide, inmate suicide, and incarcerated veterans, suggesting that the veteran in jail or prison faces a level of suicide risk beyond that conferred by either veteran status or incarceration alone. There is a clear need for a better characterization of the incarcerated veteran population and the suicide rate faced by this group. Implications for clinical practice and future research are offered. PMID- 19297639 TI - Suicide and incarcerated veterans--don't wait for the numbers. AB - Using indirect evidence, Wortzel and his colleagues raise the likelihood that the rates of suicide will increase among incarcerated veterans, given past trends and current information about veterans returning from Iraq and Afghanistan. Although it might be argued that the data are inadequate for the formulation of public policy, there is sufficient information to begin creating programs for veterans now. Wortzel and colleagues suggest screening in jails to identify veterans at risk, with increased monitoring in the first weeks of incarceration, and use of the Critical Time Intervention during important transitions. While these recommendations are worthy, a better understanding of the factors associated with suicidality may help policy-makers to support programs for returning veterans before they develop the serious problems that can lead to suicide. Also promising is the Substance Abuse and Mental Health Administration's recent funding of six jail diversion programs with a focus on veterans. PMID- 19297640 TI - Boundary violation. AB - Regulation of the interstate practice of medicine represents a challenge to state medical boards. Although laws prohibiting the unlicensed practice of medicine were originally enacted to protect the public from unqualified practitioners, they could be invoked in a whole host of common clinical situations such as calling in prescriptions to a patient in another state, giving expert testimony in another jurisdiction, or reviewing radiology films on the internet, with potentially serious criminal ramifications. In this article a recent case describing a physician's being prosecuted for the illegal practice of medicine across state lines is presented and followed by a discussion of the numerous ways in which contemporary practitioners are likely to engage in such acts. The function of state medical legislation is explored as it relates to prohibitions on interstate practice. It is suggested that states, and possibly the federal government, should devise legislative solutions to allow for the good faith intermittent practice of interstate medicine. PMID- 19297641 TI - Clinical and conceptual problems in the attribution of malingering in forensic evaluations. AB - The authors review clinical and conceptual errors that contribute to false attributions of malingering in forensic evaluations. Unlike the mental disorders, malingering is not defined by a set of (relatively) enduring symptoms or traits; rather, it is an intentional, externally motivated, and context-specific form of behavior. Despite this general knowledge, attributions of malingering are often made by using assessment tools that may detect feigning but cannot be relied upon to determine incentive and volition or consciousness (defining characteristics of malingering). In addition, forensic evaluators may overlook the possibility that feigning is a function of true pathology, as in Ganser syndrome or the factitious disorders, or that a seemingly malingered presentation is due to symptoms of an underlying disorder, such as dissociative identity disorder (DID). Other factors that set the stage for false positives, such as pressure on forensic specialists to identify malingering at all costs, failure to consider the base rate problem, and cultural variables, are also reviewed. PMID- 19297642 TI - Job security and a good episode of Scrubs. PMID- 19297643 TI - Welcome developments in UK medical humanities. PMID- 19297644 TI - Economic downturn. PMID- 19297645 TI - Change. PMID- 19297647 TI - Man-midwives. PMID- 19297648 TI - Thoughts on future doctors. PMID- 19297649 TI - Hybrid theatres: nicety or necessity? PMID- 19297650 TI - Falls and frailty: lessons from complex systems. PMID- 19297651 TI - Impact of pay for performance on quality of chronic disease management by social class group in England. AB - OBJECTIVE: To examine associations between social class and achievement of selected national audit targets for coronary heart disease (CHD), diabetes and hypertension in England before and after the introduction of a major pay for performance programme in 2004. DESIGN: Secondary analysis of 2003 and 2006 national survey data for respondents with CHD and diabetes and hypertension. SETTING: England. MAIN OUTCOME MEASURE: Achievement of national audit targets for blood pressure, blood glucose and cholesterol control. RESULTS: There were no significant differences in achievement of blood pressure targets in individuals from manual and non-manual occupational groups with diabetes (2003: 65.9% v 60.3%, 2006: 67.6% v 69.7%) or hypertension (2003: 66.2% v 66.2%, 2006: 72.8% v 71.9%) before or after the introduction of pay for performance. Achievement of the cholesterol target was also similar in individuals from manual and non-manual groups with diabetes (2003: 52.5% v 46.6%, 2006: 68.7% v 70.5%) or CHD (2003: 54.3% v 53.3%, 2006: 68.6% v 71.3%). Differences in achievement of the blood pressure target in CHD [75.8% v 84.5%; AOR 0.44 (0.21-0.90)] were evident between manual and non-manual occupational groups after the introduction of pay for performance. CONCLUSION: The quality of chronic disease management in England was broadly equitable between socioeconomic groups before this major pay for performance programme and remained so after its introduction. PMID- 19297652 TI - Trends in the epidemiology and prescribing of medication for eczema in England. AB - BACKGROUND: The prevalence of eczema, particularly in younger children, increased substantially over the second half of the 20th century. Analysis of primary healthcare data-sets offers the possibility to advance understanding about the changing epidemiology of eczema. AIM: To investigate recent trends in the recorded incidence, lifetime prevalence, prescribing and consulting behaviour of patients with eczema in England. METHODS: QRESEARCH is one of the world's largest national aggregated health databases containing the records of over nine million patients. We extracted data on all patients with a recorded diagnosis of eczema and calculated annual age-sex standardized incidence and lifetime period prevalence rates for each year from 2001-2005. We also analysed the consulting behaviour of these patients when compared with the rest of the QRESEARCH database population. The number of eczema prescriptions issued to people in England was also estimated. RESULTS: The age-sex standardized incidence of eczema was 9.58 per 1000 person-years in 2001 and increased to 13.58 per 1000 patients in 2005 (p<0.001). By 2005, eczema affected an estimated 5,773,700 (95% confidence intervals [CI] 5,754,100-5,793,400) individuals in England, who, on average, consulted a general practitioner 4.02 (95% CI 4.01-4.03) times a year. During the study period, the number of eczema related prescriptions increased by 56.6% (95% CI 56.6-56.7), so by 2005 an estimated 13,690,300 (95% CI 13,643,200-13,737,600) prescriptions were issued. CONCLUSIONS: Recorded incidence and lifetime prevalence of eczema in England continue to increase. Similar increases have also been observed in the estimated number of eczema prescriptions issued to the English population. PMID- 19297653 TI - Significant lamotrigine overdose associated with acute pancreatitis. PMID- 19297654 TI - Why animal studies are often poor predictors of human reactions to exposure. PMID- 19297655 TI - Conditional GEE for recurrent event gap times. AB - This paper deals with the analysis of recurrent event data subject to censored observation. Using a suitable adaptation of generalized estimating equations for longitudinal data, we propose a straightforward methodology for estimating the parameters indexing the conditional means and variances of the process interevent (i.e. gap) times. The proposed methodology permits the use of both time-fixed and time-varying covariates, as well as transformations of the gap times, creating a flexible and useful class of methods for analyzing gap-time data. Censoring is dealt with by imposing a parametric assumption on the censored gap times, and extensive simulation results demonstrate the relative robustness of parameter estimates even when this parametric assumption is incorrect. A suitable large sample theory is developed. Finally, we use our methods to analyze data from a randomized trial of asthma prevention in young children. PMID- 19297656 TI - Parenting children with diabetes: exploring parenting styles on children living with type 1 diabetes mellitus. AB - PURPOSE: The purpose of this study was to examine the extent to which parenting styles is associated with diabetes control in children (aged 5-12 years) with type 1 diabetes, and on child and parent quality of life. METHODS: Data were collected from a total of 216 parent and child dyads, from 4 pediatric diabetes clinics in southern Ontario, using a cross-sectional survey methodology. Each parent and child independently completed the questionnaires. The study instruments included the Parenting Dimensions Inventory, Pediatric Quality of Life (diabetes specific), and chart reviews for glycosylated hemoglobin (A1C) levels. RESULTS: The results of the study demonstrated that parenting styles were not correlated with diabetes control and were weakly correlated with quality of life. Most parents reported behaviors of authoritative or democratic parenting. The mean glycosylated hemoglobin (A1C) for children in the study was slightly above optimal target range, at 8.4%. Parental education had a weak negative correlation with diabetes control. CONCLUSIONS: Parenting styles are not associated with diabetes control and quality of life in children with type 1 diabetes. However, further research should assess the impact of the determinants of parenting on children with type 1 diabetes and quality of life. PMID- 19297658 TI - Degenerate T-cell receptor recognition, autoreactive cells, and the autoimmune response in multiple sclerosis. AB - Multiple sclerosis (MS) is the leading cause of disability in the young adult population. While the immunopathogenetic mechanisms that drive the disease have been extensively studied, the autoantigens that trigger the chronic central nervous system inflammation are still not identified. Flexibility/ degeneracy of the T-cell receptor (TCR) in antigen recognition could have a physiological role in thymic selection and the development of comprehensive TCR repertoire and protection from infections. Here, the author explores the possibility that such flexibility/degeneracy may also play a role in the induction of autoimmune diseases. Major histocompatibility complex (MHC) class II alleles of the DR2 haplotype DR2a (DRB5*0101) and DR2b (DRB1*1501) are genes associated with an increased risk for MS in Caucasian populations. Peptide binding to the MHC molecule is a prerequisite for recognition by TCRs, whereby the CD4+ T-cell response is restricted by specific MHC class II DR molecules. To selectively expand and characterize DR2-restricted T-cells with degenerate TCR (TCR(deg)), the authors designed MHC class II DR2-anchored peptide mixtures, which preferentially bind to the DR2a and DR2b antigen-presenting molecules. Peptides in these mixtures have specific amino acids in the DR2 binding positions but have randomized amino acids at all other positions of the peptide. Due to the low concentration of individual peptides in these mixtures/libraries, the authors assume that only T-cells with TCR(deg) will proliferate in response to these mixtures. The authors have recently identified an increased DR2 restricted TCR(deg) T-cell frequency in MS patients in comparison to healthy controls, their cross-reactivity to myelin basic protein, and the secretion of proinflammatory cytokines, all of which suggest that these cells may play a role in the development of the autoimmune response in MS. PMID- 19297657 TI - Colorectal cancer screening in primary care: theoretical model to improve prevalence in the practice partner research network. AB - Colorectal cancer (CRC) is the second leading cause of cancer death in the United States. Half of Americans older than age 50 are not current with recommended screening; research is needed to assess the impact of interventions designed to increase receipt of CRC screening. The Colorectal Cancer Screening in Primary Care (C-TRIP) study is a theoretically informed group randomized trial within 32 primary care practices. Baseline median proportion of active patients aged 50 years or older up-to-date with CRC screening among the 32 practices was 50.8% (N = 55,746). Men were more likely to have been screened than women (52.9% vs. 49.2%, respectively). Patients 50 to 59 years of age were less likely to be up-to date with screening (45.4%) than those in the 60 to 69 years and 70 to 79 years groups (58.5% in both groups). Opportunities exist to increase the proportion of CRC screening received in adults aged 50 and older. C-TRIP evaluates the effectiveness of a model for improvement for increasing this proportion. PMID- 19297659 TI - Differential IL-23 requirement for IL-22 and IL-17A production during innate immunity against Salmonella enterica serovar Enteritidis. AB - Early activation of the IL-12/IFN-gamma axis has been shown following Salmonella enterica serovar Enteritidis (S. Enteritidis) infection. We were interested to study whether IL-22 and IL-17A production is initiated early in response to S. Enteritidis. We demonstrate here that IL-22 was strongly elevated in the peritoneal lavage fluid and in serum already 1 day post-intraperitoneal infection (d.p.i.) of mice; not only IL-22 but also IL-17A was produced ex vivo by activated peritoneal exudate cells (PEC). Peritoneal gammadelta T cells were identified as cellular source of IL-17A. The early IL-22 production was completely IL-23-dependent. In contrast, IL-17A production was only partially IL 23-dependent. To investigate the local production of upstream cytokines important for induction of IL-22, IL-17A and IFN-gamma during salmonellosis, the production of IL-23 and IL-12 was studied. Elevated p19 and p40 mRNA levels were found in PEC at 1 d.p.i., whereas p35 mRNA levels were not changed. Besides, the T(h)17 promoting cytokines IL-6, IL-1beta and transforming growth factor-beta were produced in response to S. Enteritidis. However, IL-6 was not required for IL-22 or IL-17A production by PEC. By ex vivo analysis of PEC at 1 d.p.i., we show that the major producers of early IL-12/23p40 in the peritoneal cavity were dendritic cells (DC), whereas macrophages notably contributed to IL-6 production. Taken together, these data suggest that DC initiate early IL-22 production at the site of infection which may contribute to resistance against salmonellosis. Furthermore, we provide evidence that production of IL-22 and IL-17A is differentially regulated during infection. PMID- 19297660 TI - A question of breeding. PMID- 19297661 TI - In the shadows. PMID- 19297662 TI - Muddying the waters. PMID- 19297663 TI - Patent issues. PMID- 19297664 TI - Michael Platt. PMID- 19297665 TI - [Professionalism and quality of private respectively public treatment of eating disorders]. PMID- 19297666 TI - [Crossroads of Pathology, 17-21 November 2008, Paris, France. Proceedings and abstracts]. PMID- 19297667 TI - Abstracts from the British Association for Tissue Banking 2006 Annual Meeting, Nottingham, United Kingdom. PMID- 19297668 TI - Proceedings of the Joint International Tropical Medicine Meeting 2002. PMID- 19297671 TI - Medicaid restructuring. Issue brief. PMID- 19297672 TI - Medicaid eligibility. Issue brief. PMID- 19297673 TI - Federal Medicaid policy. Issue brief. PMID- 19297674 TI - Medicaid copayments. Issue brief. PMID- 19297675 TI - Medicaid provider tax. Issue brief. PMID- 19297676 TI - Medicaid reimbursement. Issue brief. PMID- 19297677 TI - Medicaid waivers. Issue brief. PMID- 19297678 TI - Central obesity and increased risk of dementia more than three decades later. PMID- 19297679 TI - The urinary steroid profile in patients diagnosed with adrenal incidentaloma. AB - OBJECTIVE: The aim of this study was to investigate the possible urinary markers of hormonal activity in patients with non-functioning adrenal incidentalomas. In order to evaluate the endocrine activity of aforementioned tumours, urinary steroid metabolite levels were analyzed in samples from patients and controls. Possible blocks in metabolic pathways of the examined hormones were determined by comparing selected urinary steroid metabolite sums and ratios in both groups of interest. DESIGN: Urine samples were collected from 20 patients with non functioning adrenal incidentalomas and from 25 controls matched in terms of age, sex and BMI. Excretion of 19 major urinary steroid metabolites was analyzed by gas chromatography. The results were subjected to statistical analysis. RESULTS: In patients with adrenal incidentalomas sum of total urinary cortisol metabolites was significantly increased in respect to the control group. We also observed a shift towards tetrahydrocorticosterone, cortisol and etiocholanolone production in patients. No significant differences in production of other urinary steroid metabolites were noted in patients with adrenal incidentalomas in respect to control group. CONCLUSIONS: Our data suggests that not only urinary free cortisol but also its metabolite such as tetrahydrocortisol and other steroids including etiocholanolone and corticosterone tetrahydrometabolite might be urinary markers for the endocrine activity of adrenal incidentalomas. Enhanced levels of these urinary steroid metabolites indicate an impairment of 11beta-hydroxysteroid dehydrogenase activity and slightly increased activity of 5beta-reductase in patients with adrenal incidentalomas. PMID- 19297680 TI - Medicaid services. Issue brief. PMID- 19297681 TI - Long-term care: facility quality and safety. End-of-year issue brief. PMID- 19297682 TI - Central obesity and increased risk of dementia more than three decades later. PMID- 19297683 TI - Long-term care: end-of-life issues. End-of-year issue brief. PMID- 19297684 TI - Real-time reverse transcription loop-mediated isothermal amplification for rapid and simple quantification of WT1 mRNA. AB - OBJECTIVES: This study developed a novel MRD monitoring method targeting Wilms' tumor gene (WT1) mRNA using reverse transcription loop-mediated isothermal amplification (RT-LAMP). DESIGN AND METHODS: A primer set for the assay was designed on the basis of the sequences between the 17AA and KTS regions of WT1mRNA. WT1 mRNA was quantified by real-time RT-LAMP and the accuracy of RT-LAMP was compared with that of real-time RT-PCR. RESULTS: The standard curve was expressed as a linear relationship between the log copy numbers of WT1 mRNA ranging from 6.8 x 10 to 6.8 x 10(9) copies and the threshold time with a correlation coefficient of R(2) > 0.994. The measured values obtained by RT-LAMP strongly correlated with those obtained by real-time RT-PCR. CONCLUSION: RT-LAMP can be used to determine WT1 mRNA expression levels. This assay will contribute to a more specific, simple, and rapid MRD monitoring than conventional assays. PMID- 19297685 TI - The Sequence Analysis and Management System -- SAMS-2.0: data management and sequence analysis adapted to changing requirements from traditional sanger sequencing to ultrafast sequencing technologies. AB - DNA sequencing plays a more and more important role in various fields of genetics. This includes sequencing of whole genomes, libraries of cDNA clones and probes of metagenome communities. The applied sequencing technologies evolve permanently. With the emergence of ultrafast sequencing technologies, a new era of DNA sequencing has recently started. Concurrently, the needs for adapted bioinformatics tools arise. Since the ability to process current datasets efficiently is essential for modern genetics, a modular bioinformatics platform providing extensive sequence analysis methods, is designated to achieve well the constantly growing requirements. The Sequence Analysis and Management System (SAMS) is a bioinformatics software platform with a database backend designed to support the computational analysis of (1) whole genome shotgun (WGS) bacterial genome sequencing, (2) cDNA sequencing by reading expressed sequence tags (ESTs) as well as (3) sequence data obtained by ultrafast sequencing. It provides extensive bioinformatics analysis of sequenced single reads, sequencing libraries and fragments of arbitrary DNA sequences such as assembled contigs of metagenome reads for instance. The system has been implemented to cope with several thousands of sequences, efficiently processing them and storing the results for further analysis. With the project setup, SAMS automatically recognizes the data type. PMID- 19297686 TI - Built environment and changes in blood pressure in middle aged and older adults. AB - OBJECTIVE: Few studies have examined interaction effects between person and environment, especially for cardiovascular disease (CVD) risk. The purpose of this study was to examine built environment characteristics and resident health behaviors as they relate to change in blood pressure, an important component of CVD. METHODS: Participants (N=1145, aged 50-75 at baseline) were recruited from 120 neighborhoods in Portland, Oregon. Using a longitudinal design, we assessed changes in participants' systolic and diastolic blood pressure from baseline to 1 year follow-up (2006-2007 to 2007-2008). Independent variables included baseline neighborhood-level measures of GIS-constructed neighborhood walkability and density of fast-food restaurants, and resident-level measures of meeting physical activity recommendations and eating fruits and vegetables. RESULTS: There was a small but significant resident-level increase in both systolic and diastolic blood pressure (P<0.001) over the 1-year observation period. A similar trend was also observed at the neighborhood level (P<0.001). Significant differences in change in blood pressure, by neighborhood walkability, were observed, with decreases in systolic and diastolic blood pressure for those living in high walkable neighborhoods (P<0.001). Neighborhoods of low walkability but with a high density of fast-food outlets and residents making visits to fast-food restaurants were significantly associated with increases in blood pressure measures over time. The negative effect of fast-food restaurants on blood pressure was diminished among high-walkable neighborhoods, with benefits observed among residents meeting guidelines for physical activity and eating fruits and vegetables. CONCLUSIONS: Neighborhoods with high walkability may ameliorate the risk of hypertension at the community level and promotion of neighborhood walkability could play a significant role in improving population health and reducing CVD risk. PMID- 19297687 TI - Physical activity and depressive symptoms in Taiwanese older adults: a seven-year follow-up study. AB - INTRODUCTION: The associations of leisure-time physical activity with incident depressive symptoms in a 7-year population-based sample of Taiwanese older people were studied. METHODS: The analyses were based on nationally representative data from the Taiwan's Health and Living Status of the Elderly Survey in 1996, 1999 and 2003. Data from the fixed cohort of 3778 participants aged 50 and older at baseline in 1996 with 7 years of follow-up were analyzed. Incident cases (CES-D 10 score > or =10) in 2003 totaled 420. Participants engaging in at least three activity sessions per week were classified as being physically active. Based on physical activity status in 1996 and 1999, four categories of activity status were created (low/low, low/high, high/low and high/high). Multivariate logistic regression models for predicting incident cases were undertaken both including and excluding participants with physical limitation at baseline and depressive symptoms in 1999. Sensitivity analyses were conducted to examine the effect of missing values. RESULTS: Participants who were low in leisure-time activity at both time points were at greatest risk of developing depressive symptoms (adjusted odd ratio=1.43, 95% confidence interval: 1.04-1.95) in 2003. CONCLUSIONS: Leisure-time physical activity is associated with reduced risk of significant depressive symptoms in older Taiwanese adults. PMID- 19297688 TI - CLUSEAN: a computer-based framework for the automated analysis of bacterial secondary metabolite biosynthetic gene clusters. AB - Bacterial secondary metabolites are an important source of antimicrobial and cytostatic drugs. These molecules are often synthesized in a stepwise fashion by multimodular megaenzymes that are encoded in clusters of genes encoding enzymes for precursor supply and modification. In this work,we present an open source software pipeline, CLUSEAN (CLUster SEquence ANalyzer) that helps to annotate and analyze such gene clusters. CLUSEAN integrates standard analysis tools, like BLAST and HMMer, with specific tools for the identification of the functional domains and motifs in nonribosomal peptide synthetases (NRPS)/type I polyketide synthases (PKS) and the prediction of specificities of NRPS. PMID- 19297690 TI - An evaluation framework for statistical tests on microarray data. AB - Microarray analysis has become a popular and routine method in functional genomics. It is typical for such experiments to involve a small number of replicates, which causes unreliable estimates of the sample variance. Microarrays have fostered the development of new statistical methods to analyze data resulting from experiments with small sample sizes. In this study, we tackle the problem of evaluating the performance of statistical tests for generating ranked gene lists from two-channel direct comparisons. We propose an evaluation method based on a oligonucleotide microarray with a large number of replicate spots yielding a maximum of 400 replicates per gene. We apply Spearman's rank correlation coefficient to ranked gene-lists generated by eight widely used microarray specific test statistics, which are applied to small random samples. We could show that variance stabilizing methods such as Cyber-T, SAM, and LIMMA can be beneficial for very small sample sizes and that SAM and the t-test provide stronger control of the type I error rate than the other methods. Specifically, we report that for four replicates all methods reach a high to very high correlation with our reference standard. PMID- 19297689 TI - Obesity, overweight, and weight control practices in U.S. veterans. AB - OBJECTIVES: We assessed the burden of obesity and overweight, as well as trends in weight control practices, among U.S. veterans and users of Department of Veterans Affairs (VA) health care in a large national survey. METHODS: Data were combined from the 2003 and 2004 Behavioral Risk Factor Surveillance System surveys of U.S. adults (n=558,426), a large telephone survey conducted in all states. RESULTS: Some 24% of veterans were obese (Body Mass Index [BMI] > or = 30 kg/m2), and 48% were overweight (BMI 25-29.9 kg/m2). After adjustment for age and gender, obesity prevalence was similar between veterans and non-veterans, but overweight was about 3.8 percentage points more frequent in veterans. Among veterans, obesity was more common among users of VA care, especially those who received all health care through the VA. Obese and overweight VA users were significantly more likely than non-users to report advice from a health care provider to lose weight, but no more likely to report that they were actually trying to lose weight. CONCLUSIONS: Obesity and overweight pose special burdens on veterans who use VA care. These results strengthen the case for system-wide interventions aimed at weight control. PMID- 19297691 TI - Heterologous expression, isotopic-labeling and immuno-characterization of Cin1, a novel protein secreted by the phytopathogenic fungus Venturia inaequalis. AB - The phytopathogenic fungus Venturia inaequalis causes scab of apple. Once this fungus penetrates the plant surface, it forms a specialized body called a stroma between the inner cuticle surface and the epidermal cell wall. A novel V. inaequalis gene, cin1, is strongly up-regulated in the early stages of infection. This gene codes for a 523 residue secreted protein, containing eight imperfect repeats of approximately 60 amino acids. Cin1 was expressed in the methanolytic yeast Pichia pastoris using the pPICZ vector system. A protein of 57 kDa was secreted by these transformants and peptide fingerprinting indicated that it was the Cin1 protein product. Multiple angle laser light scattering confirmed the predicted mass of Cin1, showing it was not glycosylated by Pichia and was monomeric in solution. Through measurements of the hydrodynamic properties of Cin1, the experimental Stokes radius of Cin1 was calculated and corresponded to the theoretical value for a natively folded globular protein of size 57 kDa. The mobility of recombinant Cin1 on native PAGE was also consistent with that of a folded protein. To simplify future structural analyses, a two-domain truncated version, Cin1-2D, consisting of domains one and two, was also expressed using the same vector system. Both proteins were purified to homogeneity. Conditions for maximal (>98%) incorporation of 13C and 15N were determined. A mouse polyclonal antibody and three monoclonal antibodies (MAbs) were raised against the full length version of Cin1. Analysis of the three MAbs using surface plasmon resonance indicated binding to distinct epitopes on the Cin1 protein. Western blots confirmed the different specificities of each MAb. PMID- 19297692 TI - An optimized procedure for efficient phage display of antibody fragments with a low folding efficiency. AB - We recently developed an efficient bacterial expression system for phagemid-coded antigen-binding fragments of antibody (Fabs) without the use of a helper bacteriophage. This system is characterized by an unusually long cultivation at a low temperature and gentle induction of Fab expression without the addition of the inducer isopropyl-beta-D-thiogalactopyranoside (IPTG). This method allows for a high yield production of Fabs fused with phage gene III coat protein, even when the protein is defective in its folding ability. With this cultivation procedure, we aimed here at improving the production and selection efficiency of filamentous bacteriophages displaying functional Fabs on their surface (Fab-phages) that have high affinity but low folding ability. The Fab components of the Fab-phages used were clonally related but differed in their affinity and folding ability. The production of the functional Fab-phages was quantitatively evaluated under various culture conditions. With conventional phage particle preparation, the production of functional Fab-phages was significantly biased according to the folding ability of the displayed Fabs, and affinity-based biopanning was therefore unsuccessful. In contrast, with the present procedure employing cultivation at 25 degrees C for 16 h without IPTG induction, functional Fab phages were produced without any such dependence on folding ability. With this optimized library, affinity-based biopanning was successful. Especially noteworthy, bead-based biopanning accurately discriminated between high affinity Fab-phages and Fab-phages with low or middling affinity. In obtaining Fab-phages with high affinity but low folding ability, these optimized procedures for both cultivation and selection were essential. PMID- 19297693 TI - Cloning, expression, purification and crystallization of the Rho transcription termination factor from Thermotoga maritima. AB - Rho is an essential ATP-dependent homohexameric helicase that is found in the vast majority of bacterial species. It is responsible for transcription termination at factor-dependent terminators. Rho binds to a specific region of the newly-synthesised mRNA and translocates along the chain until it reaches and disassembles the transcription complex. Basically, two crystallographic structures of Rho hexamer from Escherichia coli have been reported: an open ring with RNA (or ssDNA) bound to the RNA-binding domain, and a closed ring with the RNA bound to both the RNA-binding domain and the ATP-ase domain. The structure of the protein free from RNA is still unknown, but thermophilic bacteria enable an alternative approach to its characterization as their proteins often crystallize more easily than those of their mesophilic homologs. We report here the heterologous expression in E. coli of full-length Rho from the thermophile Thermotoga maritima, a simple protocol for the purification of its hexameric nucleic acid-free form, and the obtainment of 2.4 A-diffracting crystals. PMID- 19297694 TI - Development and crystallization of a minimal thermostabilised G protein-coupled receptor. AB - Structure determination of G protein-coupled receptors is still in its infancy and many factors affect whether crystals are obtained and whether the diffraction is of sufficient quality for structure determination. We recently solved the structure of a thermostabilised turkey beta 1-adrenergic receptor by crystallization in the presence of the detergent octylthioglucoside. Three factors were essential for this success. Firstly, truncations were required at the N-terminus to give optimal expression. Secondly, 6 thermostabilising point mutations were incorporated to make the receptor sufficiently stable in short chain detergents to allow crystallization. Thirdly, truncations at the C-terminus and within cytoplasmic loop 3, in combination with the removal of the palmitoylation site, were required to obtain well-diffracting crystals in octylthioglucoside. Here, we describe the strategy employed and the utility of thermostability assays in assessing how point mutations, truncations, detergents and ligands combine to develop a construct that forms diffraction-grade crystals. PMID- 19297695 TI - Purification and characterization of chitinase from the stomach of silver croaker Pennahia argentatus. AB - A chitinase was purified from the stomach of a fish, the silver croaker Pennahia argentatus, by ammonium sulfate fractionation and column chromatography using Chitopearl Basic BL-03, CM-Toyopearl 650S, and Butyl-Toyopearl 650S. The molecular mass and isoelectric point were estimated at 42 kDa and 6.7, respectively. The N-terminal amino acid sequence showed a high level of homology with family 18 chitinases. The optimum pH of silver croaker chitinase toward p nitrophenyl N-acetylchitobioside (pNp-(GlcNAc)2) and colloidal chitin were observed to be pH 2.5 and 4.0, respectively, while chitinase activity increased about 1.5- to 3-fold with the presence of NaCl. N-Acetylchitooligosaccharide ((GlcNAc)n, n = 2-6) hydrolysis products and their anomer formation ratios were analyzed by HPLC using a TSK-GEL Amide-80 column. Since the silver croaker chitinase hydrolyzed (GlcNAc)4-6 and produced (GlcNAc)2-4, it was judged to be an endo-type chitinase. Meanwhile, an increase in beta-anomers was recognized in the hydrolysis products, the same as with family 18 chitinases. This enzyme hydrolyzed (GlcNAc)5 to produce (GlcNAc)2 (79.2%) and (GlcNAc)3 (20.8%). Chitinase activity towards various substrates in the order pNp-(GlcNAc)n (n = 2 4) was pNp-(GlcNAc)2 >> pNp-(GlcNAc)4 > pNp-(GlcNAc)3. From these results, silver croaker chitinase was judged to be an enzyme that preferentially hydrolyzes the 2nd glycosidic link from the non-reducing end of (GlcNAc)n. The chitinase also showed wide substrate specificity for degrading alpha-chitin of shrimp and crab shell and beta-chitin of squid pen. This coincides well with the feeding habit of the silver croaker, which feeds mainly on these animals. PMID- 19297696 TI - Long-term care: home- and community-based services. End-of-year issue brief. PMID- 19297697 TI - Influence of key residues on the heterologous extracellular production of fungal ribonuclease U2 in the yeast Pichia pastoris. AB - Ribonuclease U2, secreted by the smut fungus Ustilago sphaerogena, is a cyclizing ribonuclease that displays a rather unusual specificity within the group of microbial extracellular RNases, best represented by RNase T1. Superposition of the three-dimensional structures of RNases T1 and U2 suggests that the RNase U2 His 101 would be the residue equivalent to the RNase T1 catalytically essential His 92. RNase U2 contains three disulfide bridges but only two of them are conserved among the family of fungal extracellular RNases. The non-conserved disulfide bond is established between Cys residues 1 and 54. Mispairing of the disulfide network due to the presence of two consecutive Cys residues (54 and 55) has been invoked to explain the presence of wrongly folded RNase U2 species when produced in Pichia pastoris. In order to study both hypotheses, the RNase U2 H101Q and C1/54S variants have been produced, purified, and characterized. The results obtained support the major conclusion that His 101 is required for proper protein folding when secreted by the yeast P. pastoris. On the other hand, substitution of the first Cys residue for Ser results in a mutant version which is more efficiently processed in terms of a more complete removal of the yeast alpha-factor signal peptide. In addition, it has been shown that elimination of the Cys 1-Cys 54 disulfide bridge does not interfere with RNase U2 proper folding, generating a natively folded but much less stable protein. PMID- 19297698 TI - Optimized procedures for producing biologically active chemokines. AB - We describe here two strategies to produce biologically active chemokines with authentic N-terminal amino acid residues. The first involves producing the target chemokine with an N-terminal 6xHis-SUMO tag in Escherichia coli as inclusion bodies. The fusion protein is solubilized and purified with Ni-NTA-agarose in denaturing reagents. This is further followed by tag removal and refolding in a redox refolding buffer. The second approach involves expressing the target chemokine with an N-terminal 6xHis-Trx-SUMO tag in an engineered E. coli strain that facilitates formation of disulfide bonds in the cytoplasm. Following purification of the fusion protein via Ni-NTA and tag removal, the target chemokine is refolded without redox buffer and purified by reverse phase chromatography. Using the procedures, we have produced more than 15 biologically active chemokines, with a yield of up to 15 mg/L. PMID- 19297699 TI - Access to health insurance: State Childrens's Health Insurance Program (SCHIP). Issue brief. PMID- 19297700 TI - Medical errors and patient safety. Issue brief. PMID- 19297701 TI - CBM21 starch-binding domain: a new purification tag for recombinant protein engineering. AB - The use of protein fusion tag technology simplifies and facilitates purification of recombinant proteins. In this article, we have found that the starch-binding domain derived from Rhizopus oryzae glucoamylase (RoSBD), a member of carbohydrate-binding module family 21 (CBM21) with raw starch-binding activity, is favorable to be applied as an affinity tag for fusion protein engineering and purification in Escherichia coli and Pichia pastoris systems. To determine suitable spatial arrangement of RoSBD as a fusion handle, enhanced green fluorescent protein (eGFP) was fused to either the N- or C-terminus of the SBD, expressed by E. coli, and purified for yield assessment and functional analysis. Binding assays showed that the ligand-binding capacity was fully retained when the RoSBD was engineered at either the N-terminal or the C-terminal end. Similar results have been obtained with the RoSBD-conjugated phytase secreted by P. pastoris. The effective adsorption onto raw starch and low cost of starch make RoSBD practically applicable in terms of development of a new affinity fusion tag for recombinant protein engineering in an economic manner. PMID- 19297702 TI - Medical malpractice and tort reform. Issue brief. PMID- 19297703 TI - Mandated benefits. Issue brief. PMID- 19297704 TI - Managed care. Issue brief. PMID- 19297705 TI - Quality of healthcare. Issue brief. PMID- 19297706 TI - Healthcare information technology. Issue brief. PMID- 19297707 TI - Healthcare workforce. Issue brief. PMID- 19297708 TI - Health facilities. Issue brief. PMID- 19297709 TI - 2008 IADR Pre-Prosthetic Regenerative Science Award. PMID- 19297710 TI - [Multidisciplinary approach to the diagnosis of Takotsubo cardiomyopathy]. PMID- 19297711 TI - Re: Postoperative fluid balance in patients having operations on the head and neck. PMID- 19297712 TI - Manitoba passes forced testing legislation. PMID- 19297713 TI - Alberta to make personal medical information available on-line. PMID- 19297714 TI - WHO commission reports on social determinant's of health. PMID- 19297715 TI - Court dismisses Trevis Smith's appeal from his conviction on two charges of aggravated sexual assault. PMID- 19297716 TI - Commentary by Hinck. PMID- 19297717 TI - Pharmaceuticals and medical devices: cost savings. PMID- 19297718 TI - Pharmaceuticals and medical devices: FDA oversight. PMID- 19297719 TI - Pharmaceuticals and medical devices: business practices. PMID- 19297720 TI - Pharmaceuticals and medical devices: medical devices. PMID- 19297721 TI - Access to health insurance. PMID- 19297722 TI - Involvement of reactive nitrogen and oxygen species (RNS and ROS) in sunflower mildew interaction. Plant Cell Physiol. 50(2): 265-79 (2009). PMID- 19297724 TI - Biotechnology for the sustainability of human society. Abstracts of IBS 2008, the 13th International Biotechnology Symposium and Exhibition. Dalian, China. October 12-17, 2008. PMID- 19297723 TI - A technique for controlled needle decompression of distended bowel. PMID- 19297725 TI - Enzymatic hydrolysis and fermentability of corn stover pretreated by lactic acid and/or acetic acid. AB - Four different pretreatments with and without addition of low concentration organic acids were carried out on corn stover at 195 degrees C for 15 min. The highest xylan recovery of 81.08% was obtained after pretreatment without acid catalyst and the lowest of 58.78% after pretreatment with both acetic and lactic acid. Glucan recovery was less sensitive to the pretreatment conditions than xylan recovery. The pretreatment with acetic and lactic acid yielded the highest glucan recovery of 95.66%. The glucan recoveries of the other three pretreatments varied between 83.92% and 94.28%. Fermentability tests were performed on liquors obtained from all pretreatments and there were no inhibition effect found in any of the liquors. Simultaneous saccharification and fermentation (SSF) of water insoluble solids (WIS) showed that a high ethanol yield of 88.7% of the theoretical based on glucose in the raw material was obtained following pretreatment at 195 degrees C for 15 min with acetic acid employed. The estimated total ethanol production was 241.1 kg/ton raw material by assuming fermentation of both C-6 and C-5, and 0.51 g ethanol/g sugar. PMID- 19297726 TI - Application of aqueous two-phase systems to antibody purification: a multi-stage approach. AB - The single-stage equilibrium aqueous two-phase extraction (ATPE) of human antibodies from a Chinese hamster ovary (CHO) cells supernatant was investigated. The performance of polyethylene glycol 3350 (PEG 3350)/phosphate aqueous two phase systems (ATPS) was evaluated by studying several experimental conditions, such as pH, ionic strength, volume ratio and initial antibody concentration in the feed stock. The conditions that favoured the extraction of human immunoglobulin G (IgG) were low pH values, high NaCl concentration and high volume ratios. The percentage of contaminants removal did not depend on the pH value and NaCl concentration, increasing, however, as the volume ratio decreased. About 66% of total contaminants were removed when a volume ratio of 0.4 was used. The multi-stage equilibrium ATPE was also investigated by simulating a four stages cross-current operation in test tubes. According to the IgG equilibrium curves and respective McCabe Thiele diagrams, a predicted optimised scheme of a counter-current multi-stage ATPE was described. An IgG recovery yield of 89% and a protein purity of 75% can be achieved using a PEG/phosphate ATPS containing 10% (w/w) NaCl, five stages and a volume ratio of 0.4. This represents significant improvements in the recovery yield and purity when compared to a single-stage trial performed at the same experimental conditions, where a 61% recovery yield and 55% protein purity were attained. Based on the CHO cells supernatant components equilibrium curves, it was observed that IgG can be completely purified from the higher molecular weight (MW) components and partially purified from the lower MW components. PMID- 19297727 TI - A novel hybrid dual-channel catalytic-biological sensor system for assessment of fruit quality. AB - The release of volatile ethylene and acetaldehyde characterizes the metabolic state and quality of fruit. We have designed and implemented a hybrid dual channel catalytic-biological sensor system, which is able to quantify both volatiles in situ. This sensor system consists of a mammalian cell line engineered for constitutive expression of an Aspergillus nidulans-derived biosensor which triggers quantitative reporter gene expression in the presence of volatile acetaldehyde. Ethylene, oxidized to acetaldehyde using a Wacker-based process, can be quantified by the same transgenic sensor cell line. Differential profiling of reporter gene transcription by the sensor system revealed the relative concentrations of both volatile metabolites and enabled correct assessment of fruit quality as shown for fresh, old and rotten apples. Functional combination of catalytic processes with biosensor technology is able to precisely capture the metabolic state of food and may foster novel insight into biochemical food quality assessment as well as the design of synthetic control circuits detecting and preventing food spoilage. PMID- 19297728 TI - High-level production of a candidacidal peptide lactoferrampin in Escherichia coli by fusion expression. AB - Expression of lactoferrampin 265-284 (Lfampin20), a potential candidacidal agent with 20 amino acid segment from lactoferrin, in Escherichia coli was explored. The DNA fragment encoding Lfampin20 was synthesized in light of the E. coli preferred codons by "partially overlapping primer-based PCR" method. The Lfampin20 gene was fused with thioredoxin (Trx) gene to construct a recombinant plasmid pETLfa20. The resulting expression level of the fusion protein Trx Lfampin20 (approximately 20 kDa) accounted for 34-42% of cellular protein, and about 52% of the target proteins were in a soluble form. Soluble Trx-Lfampin20 accounted for 66% of the total soluble proteins. The soluble fusion protein was easily purified to near homogeneity by affinity chromatography using hexahistidine tag. Recombinant Lfampin20 was effectively obtained by on-column cleavage of the fusion protein with factor Xa. An unknown site in the Trx-tag fusion protein, which can be cleaved by factor Xa to produce approximately 10 kDa protein, was found. Compared with the unknown site, the specific site of IEGR[downwards arrow]X was easier to be recognized and cleaved by factor Xa. The molecular mass of recombinant Lfampin20 determined by MALDI-TOF (matrix assisted laser desorption ionization-time-of-flight) is equal to its theoretical molecular weight. Antimicrobial activity assays demonstrated that the recombinant Lfampin20 had candidacidal activity. Integration of the key strategies for the expression of antimicrobial peptides (AMPs) such as codon usage bias, fusion partner and on column cleavage, would provide an efficient and facile platform for the production or study of AMPs. PMID- 19297729 TI - Itch-related responses of dorsal horn neurons to cutaneous allergic stimulation in mice. AB - This study was conducted to identify the mechanosensitive dorsal horn neurons involved in allergic itch. We examined 98 units responsive to cutaneous allergy; 90 showed only immediate responses, which subsided before the onset of itch related behavior and eight showed immediate and sustained responses, the latter of which was similar in duration to itch-related behavior, suggesting the involvement of sustained units in itch signaling. Sustained units were localized in the superficial, but not deep, layers of the dorsal horn. They were wide dynamic range or nociceptive specific, but not low threshold and four of eight were noxious heat sensitive. The results suggest that a small minority of neurons in the superficial dorsal horn are involved in allergic itch signals. PMID- 19297730 TI - World update: Mumbai attacks strain relations. PMID- 19297731 TI - Event-related potential correlates of spatiotemporal regularities in vision. AB - Spatiotemporal regularities in stimulus structure have been shown to influence visual target detection and discrimination. Here, we investigate whether the influence of spatiotemporal regularities is associated with the modulation of early components (P1/N1) in event-related potentials. Stimuli consisted of five horizontal bars (predictors) appearing successively towards the fovea followed by a target bar at fixation, and participants performed a key-press on target detection. Results showed that compared with the condition where five predictors were presented in a temporally regular but spatially randomized order, target detection times were faster and contralateral N1 peak latencies were shorter when the predictors and the target were presented with spatial and temporal regularities. Both measures were most prolonged when only the target was presented. In this latter condition, an additional latency prolongation was observed for the P1 peak compared with the conditions where the target was preceded by the predictors. The latency shifts associated with early event related potential components provide additional support for the involvement of early visual processing stages in the coding of spatiotemporal regularities in humans. PMID- 19297732 TI - Internet for patients. PMID- 19297733 TI - IT challenges. PMID- 19297734 TI - Imaging. New radiology service copies the eBay model. PMID- 19297735 TI - Keep quiet about a sale? PMID- 19297737 TI - Angels and pins. PMID- 19297736 TI - Technology. Executive, nurse involvement key to a successful IT launch. PMID- 19297738 TI - Mental representation of space in vestibular patients with otolithic or rotatory vertigo. AB - In previous studies, we had observed that the occurrence of geometric illusions was reduced when healthy observers were tilted relative to gravity or placed in microgravity. We hypothesized that the alteration of the gravitational (otolith) input was responsible for this change, presumably because of a connection between vestibular and visual-spatial cognitive functions. In this study, we repeated these experiments in vestibular patients who presented signs of otolith disorders. In agreement with the microgravity data, geometric illusions based on horizontal, vertical, and oblique lines were less frequent in patients with otolithic (nonrotatory) vertigo than in patients with rotatory vertigo and in healthy participants. Other visual illusions not based on perspective were not significantly different across all participant groups. We conclude that the impairment in the processing of gravitational input in the otolithic patients could be at the origin of a deformed mental representation of personal and extrapersonal space. PMID- 19297739 TI - Dissociable neural correlates of reorienting within versus across visual hemifields. AB - Neural correlates of reorienting across visual hemifields have been extensively studied, however, those of reorienting within hemifields and if there are any differences remain unclear. Here, we performed a functional magnetic resonance imaging study to identify neural correlates of reorienting within and across hemifields using a variant of the cueing paradigm. Behavioral results showed that reorienting across hemifields showed significant validity effect, but reorienting within hemifields did not. Functional magnetic resonance imaging data revealed dissociable activations in the right posterior parietal region between reorienting within and across hemifields. The present results suggest that reorienting within hemifields differs from the 'classical reorienting' to some extent, whereas reorienting across hemifields does not. PMID- 19297740 TI - Astrocytic necrosis is induced by anti-aquaporin-4 antibody-positive serum. AB - Neuromyelitis optica (NMO) is an inflammatory demyelinating disease of the central nervous system, and humoral immunity is suggested to play an important role in the pathogenesis. The identification of an anti-aquaporin-4 antibody (AQP4-Ab, neuromyelitis optica immunoglobulin G) in the sera of patients with NMO has led to the investigation on the pathogenicity of the autoantibody. Recent immunohistological analyses revealed the primary loss of AQP4 on astrocytes and complement deposition in active lesions of NMO. In this report, we show that astrocytes are susceptible to sera from AQP4-Ab-positive patients and undergo necrosis in a complement-dependent manner. Our results suggest the primary pathogenic role of AQP4-Ab in NMO. PMID- 19297741 TI - Comparison of visual evoked-related potentials in healthy young adults of different catechol-O-methyltransferase genotypes in a continuous 3-back task. AB - We further support the notion that the catechol-O-methyltransferase (COMT) polymorphism may play a role in cognitive function. A continuous visual 3-back numerical working memory task was performed for an hour among 21 patients with different COMT genotypes. P300 visual event-related potentials evoked from the 3 back task were first utilized to observe the relationship between this COMT polymorphism and cortical physiology. The results showed that Val/Met heterozygous individuals show poorer performance, lower P300 amplitude, and higher P300 latency than those bearing the Val/Val homozygote on the task. This visual 3-back paradigm is a promising tool to examine continuous working memory capacity. The role of the COMT gene in dopamine catabolism makes for the influence of COMT polymorphism on P300 endophenotypes. PMID- 19297742 TI - Should women with normal cholesterol be taking a statin? A major study suggests that statins also quell inflammation. Now what? PMID- 19297743 TI - Hypnosis helps reduce hot flashes in breast cancer survivors. PMID- 19297744 TI - Research finds that many older people can't get up from a fall. PMID- 19297745 TI - Recognizing and treating Morton's neuroma. PMID- 19297746 TI - Have your vaccinations had a check-up lately? Don't assume vaccinations received during childhood will protect you for the rest of your life. PMID- 19297747 TI - By the way, doctor. I had a laparoscopic supracervical hysterectomy a year ago. Six months ago, I started having severe pelvic pain. It seems to develop a few days before I would normally have had a menstrual period (I still have both ovaries). Is it possible to experience symptoms of endometriosis after a hysterectomy? I also wonder if I might have pelvic adhesions and this is causing the pain. PMID- 19297748 TI - HIV-positive man pleads guilty to two counts of attempted aggravated sexual assault. PMID- 19297749 TI - Winnipeg man convicted of aggravated sexual assault for failing to disclose his HIV status to former girlfriends. PMID- 19297750 TI - Court dismisses Ottawa's application to uphold federal medical marijuana restrictions. PMID- 19297751 TI - Supreme Court of India approves government commitments on health care for people living with HIV. AB - In August 2008, the Government of India issued a list of directives with respect to its national response to HIV/AIDS. A panel of the Supreme Court approved the directives on 1 October 2008 and directed that all state governments comply with them. The panel's order also directed that the National AIDS Control Organisation (NACO) submit a progress report on compliance within four months. As a result, a legally binding framework for healthcare and treatment of people living with HIV now exists for all of India. PMID- 19297752 TI - European Court rejects Ugandan woman's claim to stay in the U.K. AB - In May 2008, the Strasbourg Court issued its judgment in the case of an HIV positive woman who sought to stay in the U. K., where she was receiving antiretroviral treatment. While treatment may be inaccessible in Uganda, the Court ruled that the humanitarian grounds against removal were not compelling enough to find that her removal would constitute inhuman or degrading treatment. PMID- 19297753 TI - South African court bans promotion of vitamin treatments as cures for AIDS. AB - On 13 June 2008, the High Court of South Africa (Cape of Good Hope Provincial Division) ruled against a producer of alternative remedies and the Government of South Africa in a case regarding vitamin supplements being marketed as treatments for HIV/AIDS. The court found that the vitamin supplements were "medicines" and hence subject to regulation under the Medicines and Related Substances Act 101 of 1965. PMID- 19297754 TI - South African Court: Military cannot exclude HIV-positive people. AB - On May 16th, 2008, the High Court of South Africa ruled that the South African National Defence Force (SANDF) HIV testing policy is unconstitutional because it excludes people living with HIV from recruitment, promotion or foreign deployment. PMID- 19297755 TI - Australia: man found not guilty of deliberate HIV transmission. PMID- 19297756 TI - Finland: ten-year sentence for HIV transmission and exposure. PMID- 19297757 TI - U.S.: prison sentences for spitting. PMID- 19297759 TI - Egypt: Sentences upheld for men convicted of "debauchery". PMID- 19297758 TI - Kyrgyzstan: nine health care workers guilty of negligence causing HIV transmission among children. PMID- 19297760 TI - Human rights and HIV/AIDS: where are we? and what next? AB - Issues related to HIV/AIDS and human rights received more attention at AIDS 2008 than ever before at an International AIDS Conference. Nevertheless, in this presentation at one of two sessions devoted to HIV and human rights, Ralf Jurgens warned that despite much rhetoric, real action on HIV/AIDS and human rights remains lacking. He suggested that much needed to change before human rights will be where they need to be--at the centre of the global AIDS struggle. And he proposed a number of concrete activities to get there. PMID- 19297761 TI - Rights and policy front and centre at the conference. AB - There were more sessions than ever on human rights and policy at this conference- 86 abstracts, 46 poster discussions, 702 posters, and numerous crosscutting, bridging and satellite sessions. In this article, based on her summary of Track E (Policy and Politics) at the rapporteur session on the final day of the conference, Mandeep Dhaliwal presents a summary of key themes and messages that emerged from Track E. PMID- 19297762 TI - Lessons from Africa: combating the twin epidemics of domestic violence and HIV/AIDS. AB - Domestic violence and HIV/AIDS have proven a lethal combination, exacting a heavy toll on women's lives, particularly in Africa. In this article, partially based on a presentation made at the human rights networking zone at the conference, Tamar Ezer examines the interrelation between domestic violence and HIV/AIDS, provides an analysis of obligations under human rights law, and describes innovative programs that attempt to address the intersection of these twin epidemics. The author argues for holistic approaches that address the social, economic and legal dimensions of the problem. PMID- 19297763 TI - Sexual assault, domestic violence and HIV: promoting women's rights through legislation. AB - General HIV laws seldom, if ever, address the human rights abuses that most affect women, particularly rape, sexual assault and domestic violence. In this article, which is based on his presentation at a concurrent session at the conference, Richard Pearshouse describes a Canadian HIV/AIDS Legal Network project to develop draft legislation covering certain areas of women's rights. The draft legislation is intended to be used as a practical resource for bringing about concrete law reform. This presentation won for Richard the International AIDS Society's Young Investigator Award for the conference's Track E (Policy and Political Sciences). PMID- 19297764 TI - HIV is a virus, not a crime. AB - Criminalization of HIV transmission is an ineffective tool for combating AIDS and a costly distraction from programmes that we know work--programmes such as effective prevention, protection against discrimination, reducing stigma, empowering women and providing access to testing and treatment. In this article, which is based on a plenary presentation by Edwin Cameron, the authors advance ten reasons why criminalization is poor public health policy. PMID- 19297765 TI - Pretrial detention: scale and relevance to HIV/AIDS. AB - The overuse of pretrial detention worldwide has created conditions in detention facilities that expose large numbers of people to health risks, including the risk of contracting HIV. In this article, based on her presentation at a concurrent session at the conference, Denise Tomasini-Joshi describes the problems, and puts forward suggestions for how they can be addressed. PMID- 19297766 TI - Central & Eastern Europe and Central Asia: police raids and violence put sex workers at risk of HIV. AB - In most of the countries of Central & Eastern Europe and Central Asia, sex workers report very high levels of abuse from police, particularly in the context of police raids. In this article, based on an oral presentation at the conference, the Sex Workers' Rights Advocacy Network (SWAN) report on the results of a study conducted among sex workers in several countries in the region. The authors make the link between police violence and general violence, and between violence against sex workers and vulnerability to HIV infection. The authors advance a number of recommendations, including that sex work be decriminalized. PMID- 19297767 TI - Violence against sex workers by police and military in Democratic Republic of Congo. AB - Sex workers in the Sud-Kivu district of the Democratic Republic of Congo (DRC) are regularly subjected to sexual and other forms of violence. In this article, based on a presentation at a concurrent session at the conference, Alphonse Mihigo Ombeni and Anna Louise Crago describe the negative impacts of this violence on the sex workers' health and working conditions. Many have become HIV positive. PMID- 19297768 TI - People with disabilities and the AIDS pandemic: making the link. AB - People with disabilities are at a heightened risk of contracting HIV, yet they are not being reached by prevention and treatment campaigns. In this article, based on a presentation made at a concurrent session at the conference, Shonali Shome and Myroslava Tataryn examine the ways people with disabilities are excluded from HIV/AIDS programs; what is being done to address the gap; and what remains to be done. PMID- 19297769 TI - Coerced and forced sterilization of HIV-positive women in Namibia. AB - Thirteen cases of coerced and forced sterilization of HIV-positive women have been documented in Namibia. In this article, based on a presentation at a concurrent session at the conference, Jennifer Mallet and Veronica Kalambi explain that this is just the tip of the iceberg. The authors provide details of the 13 cases and describe how this procedure violates numerous laws and international obligations. At least two of the women have filed lawsuits in the High Court. PMID- 19297770 TI - Using strategic litigation to strengthen rights in southern Africa. AB - Litigation can be a useful tool for achieving policy and legal change, but only if accompanied by broader advocacy and education efforts. In this article, based on her oral poster presentation at the conference, Priti Patel describes the approach of the HIV/AIDS Programme at the Southern Africa Litigation Centre (SALC). PMID- 19297771 TI - Civil society strategy for the compulsory licensing of lopinavir/ritonavir: the Brazilian case. AB - The flexibilities in the TRIPS Agreement (Trade Related Aspects of Intellectual Property) have been very useful in lowering the prices of antiretrovirals (ARVs) in Brazil. In this article, based on several presentations made at the conference, Marcela Fogaca Vieira et al describe recent developments in Brazil, including the granting of a compulsory licence for efavirenz. PMID- 19297772 TI - Abuses of women's rights in sexual and reproductive health-care settings. AB - For many years, the intersection between HIV/AIDS and sexual and reproductive rights focused on the prevention of the epidemic. The violations to reproductive rights that HIV positive women face were not visible. However, this has begun to change. In this article, which is based on her presentation in the Human Rights Networking Zone at the conference, Ximena Andion Ibanez describes six areas where women's reproductive rights have been violated. The author advocates the use of litigation as a tool for advancing these rights. PMID- 19297773 TI - Drawing conceptual linkages: property rights and HIV. AB - An understanding of the impact of HIV/AIDS epidemic on broader social development necessitates a closer scrutiny of the relationship between HIV/AIDS and economic productivity. In this article, which is based on her presentation at a concurrent session at the conference, Priya Nanda describes the relationship between the widespread exclusion in developing countries of land ownership by women and their vulnerability to HIV and the effects of HIV. The author calls for more research on the complexity and diversity of land tenure systems and property laws. PMID- 19297774 TI - Measuring for change: a new research initiative by and for people living with HIV. AB - Several organizations have banded together to create the People Living with HIV Stigma Index. In this article, which is based on a presentation at a concurrent session at the conference, Lucy Stackpool-Moore and Anandi Yuvaraj describe the purpose of the index and how it was developed. The authors believe that the index provides a real opportunity to measure, understand and advocate effectively to improve policies and programs and to make a real difference in the lives of people living with HIV. PMID- 19297775 TI - Continued challenge posed by HIV-related restrictions on entry, stay and residence. AB - Restrictions on entry, stay, and residence for people living with HIV/AIDS violate international human rights law and cannot be justified by public health rationales. In this article, based upon his presentation in a concurrent session at the conference, Joe Amon argues that governments must immediately repeal these laws and policies. PMID- 19297776 TI - Putting the right back into sexual and reproductive rights. AB - There are numerous barriers to the achievement of sexual and reproductive health rights (SRHR), particularly for people living with HIV (PLHIV). In this article, which is based on his presentation at a concurrent session at the conference, Shaun Mellors, who is living with HIV, discusses the need for PLHIV to help lead efforts to ensure that their SRHR are addressed. The author also describes specific actions that donors can take to support the SRHR of PLHIV. PMID- 19297777 TI - Yogyakarta Principles: applying existing human rights norms to sexual orientation and gender identity. AB - International human rights apply to all people, including lesbian, gay, bisexual and transgender persons. But up until recently, there was no document that explained these rights specifically as they relate to sexual orientation and gender identity. In this article, based on his presentation at a concurrent session at the conference, Boris Dittrich describes the development of the Yogyakarta Principles, which not only list the rights involved, but also provide recommendations to states (and others) concerning how the rights should implemented. PMID- 19297778 TI - Rejecting the evidence: Russia's drug dependence treatment system. AB - In at least three regions of Russia, the drug treatment system fails to respond to the needs of injection drug users seeking treatment for their addiction. In this article, based on his presentation at a concurrent session at the conference, Diederik Lohman describes the results of research conducted by Human Rights Watch (HRW). The author explains that the Russian medical establishment ignores overwhelming scientific evidence that treatment substitution programs are both safe and effective. PMID- 19297779 TI - U.N. guidance note on HIV and sex work "reworked" by activists. AB - In 2007, UNAIDS issued a guidance note on HIV and sex work, the tone and contents of which angered sex workers, activists and public health workers worldwide. In this article, based on presentations at the conference, M. Seshu et al describe the problems with the guidance note, discuss the reaction to its publication, and explain how a group of activists got together to develop a reworked version of the guidance note. PMID- 19297780 TI - Recognize sex work as legitimate work. AB - It is not sex work per se that makes sex workers vulnerable to HIV, but rather the policies that repress them. In this article, based on her presentation at a plenary session at the conference, Elena Reynaga, who is a sex worker, describes how these policies deprive sex workers of their rights and subject them to physical and sexual violence. The author concludes that at the heart of the problem lies the fact that sex work is not recognized as legitimate work. PMID- 19297781 TI - [Investigations of the coroners of the district of Quebec, 1765-1930: a source in Candian medical and social history]. AB - The coroner's office, created in England in 1194 to hold inquiries into suspicious or violent deaths, was established in Quebec and Montreal in 1764. From 1765 to 1930, more than 16,000 reports, housed in the provincial National Library and Archives, were documented for the district of Quebec. A database was created to facilitate access for those interested in past customs, in provision of medical care, in attitudes towards death, in crime, in the effects of industrialization, and in new ways of production. This paper summarizes the most important elements of this database, gives a chronological account of the office of the coroner, describes the documents, and show the connection between the inquiries of the coroners and the socioeconomic events of the region. PMID- 19297782 TI - [The black death in Christian and Muslim Occident, 1347-1353]. AB - Between the years 1346 and 1353, a terrible epidemic swept over Western Asia, the Middle East, Africa, and Europe, causing catastrophic losses of population everywhere, both in the rural areas and in towns and cities. The Black Death was a disease of such magnitude that it not only shook the Old World to its economic and social foundations but changed the course of human history. The authors considered and analyzed many studies on the Black Death published in different languages. In the present paper they report medical and epidemiological specificities of the pandemic as well as its geographical origins and the routes of its spread. PMID- 19297783 TI - [Ophthalmia neonatorum of the newborn and its treatments in Canadian medical publications: 1872-1985]. AB - In the 19th century the occurrence of ophthalmia neonatorum had reached alarming rates in the maternity wards not only of Europe but also across Canada. The impact of this blinding ocular infection on Canadian medicine from 1872 to 1985 is examined through a review of 80 medical journals, books, and lay press articles of that period. The prophylactic and therapeutic use of 2% silver nitrate introduced by Crede in 1880 to prevent neonatal blindness is reviewed. The signs, symptoms, and corneal complications of this disease as well as the multiple ocular drugs used during this era will be presented. The judicial consequences on midwives and obstetricians will be discussed. The subsequent use of colloidal silver based agents such as collargol, protargol and argyrol followed by the introduction of sulfonamides and finally the routine use of prophylactic topical antibiotics will be reviewed. PMID- 19297784 TI - Black holes in bone - irresistible attractors of foreign materials? PMID- 19297785 TI - Outcome after tantalum rod implantation for treatment of femoral head osteonecrosis: 26 hips followed for an average of 3 years. AB - BACKGROUND AND PURPOSE: Tantalum rod implantation has recently been proposed for treatment of early stages of femoral head osteonecrosis.The purpose of our study was to report the early results of its use in pre- and post-collapse stages of the disease. METHODS: We studied prospectively 27 patients who underwent tantalum rod implantation for treatment of nontraumatic femoral head osteonecrosis between December 2000 and September 2005. Patients were evaluated radiologically and clinically using the Steinberg classification and the Harris hip score (HHS). Disease stage varied between stages II and IV. Mean follow-up time was 38 (15-71) months. RESULTS: 1 patient (1 hip) died 15 months after surgery for reasons unrelated to it. 13 of 26 hips remained at the same radiographic stage, and 13 deteriorated. Mean HHS improved from 49 to 85. 6 patients required conversion to total hip arthroplasty. When the procedure was used for stages III and IV, both radiological outcome and revision rates were worse than for the stage II hips. There was, however, no difference in postoperative HHS between patients at pre- and post-collapse stages at the time of initial evaluation. Survivorship, with revision to THA as the endpoint,was 70% at 6 years. INTERPRETATION: The disease process does not appear to be interrupted,but there was a significant improvement in hip function initially in most hips. Tantalum rod implantation is a safe "buy time" technique, especially when other joint salvage procedures are not an option. Appropriate patient selection and careful rod insertion are needed for favorable results. PMID- 19297786 TI - Pneumatic wound compression after hip fracture surgery did not reduce postoperative blood transfusion: A randomized controlled trial involving 292 fractures. AB - BACKGROUND AND PURPOSE: Patients with fracture of the proximal femur often undergo blood transfusion. A pneumatic compression bandage has been shown to reduce transfusion after primary hip arthroplasty for osteoarthritis. In this randomized trial, we evaluated the efficacy of this bandage following surgery for hip fracture. PATIENTS AND METHODS: 288 patients, 50 years or older with 292 fractured hips treated with hemiarthroplasty or internal fixation (except pinning), were randomized to an experimental group with pneumatic wound compression applied after surgery (n = 138) and a control group with the same dressing but no compression (n = 154). Transfusion threshold was blood hemoglobin below 100 g/L. The primary outcome measures were the number of blood units and the proportion of patients transfused after surgery. RESULTS: The primary outcome measures were similar in both groups. The mean number of postoperatively transfused blood units was 1.3 in the compression group and 1.1 in the non compression group. Blood transfusion was given to 84 patients (62%) in the compression group and to 85 patients (55%) in the non-compression group. INTERPRETATION: Pneumatic wound compression does not reduce the need for transfusion after hip fracture surgery. PMID- 19297787 TI - Periprosthetic infection: where do we stand with regard to Gram stain? AB - BACKGROUND AND PURPOSE: One of the routinely used intraoperative tests for diagnosis of periprosthetic infection (PPI) is the Gram stain. It is not known if the result of this test can vary according to the type of joint affected or the number of specimen samples collected. We examined the role of this diagnostic test in a large cohort of patients from a single institution. MATERIALS AND METHODS: A positive gram stain was defined as the visualization of bacterial cells or "many neutrophils" (> 5 per high-power field) in the smear. The sensitivity, specificity, and predictive values of each individual diagnostic arm of Gram stain were determined. Combinations were performed in series, which required both tests to be positive to confirm infection, and also in parallel, which necessitated both tests to be negative to rule out infection. RESULTS: The presence of organisms and "many" neutrophils on a Gram smear had high specificity (98-100%) and positive predictive value (89-100%) in both THA and TKA. The sensitivities (30-50%) and negative predictive values (70-79%) of the 2 tests were low for both joint types. When the 2 tests were combined in series, the specificity and positive predictive value were absolute (100%). The sensitivity and the negative predictive value improved for both THA and TKA (43-64% and 82%, respectively). INTERPRETATION: Although the 2 diagnostic arms of Gram staining can be combined to achieve improved negative predictive value (82%), Gram stain continues to have little value in ruling out PPI. With the advances in the field of molecular biology, novel diagnostic modalities need to be designed that can replace these traditional and poor tests. PMID- 19297788 TI - Once-weekly oral medication with alendronate does not prevent migration of knee prostheses: A double-blind randomized RSA study. AB - BACKGROUND AND PURPOSE: Early migration of joint replacements is an effect of poor fixation and can predict late loosening. By reducing the bone resorption after implantation of a joint replacement, it should be possible to enhance the initial fixation of the implant. We studied the effect of once-weekly treatment with alendronate after knee replacement. PATIENTS AND METHODS: We recruited 60 patients (60 knees) with gonarthrosis who were scheduled for a total knee replacement. They were operated on with identical implants and uncemented fixation. 30 patients were treated with a bisphosphonate (alendronate) and 30 patients underwent placebo treatment. The treatment started postoperatively and continued on a weekly basis for 6 months. The fixation of the implants was measured with repeated radiostereometry for 2 years. RESULTS: There was no difference in migration of implants between the two groups. CONCLUSION: With uncemented fixation of knee implants, no benefit of once-weekly treatment with alendronate, starting postoperatively, could be seen during a 2-year follow-up period. PMID- 19297789 TI - Injuries as a result of treatment of tibial fractures in children: Claims for compensation submitted to the Patient Insurance Center in Finland. AB - BACKGROUND AND PURPOSE: Tibial fractures comprise 10% of all fractures in children. To our knowledge there have been no previous reports of treatment injuries in these fractures. We analyzed compensation claims concerning treatment of these fractures in Finland. We used this information to determine preventable causes of treatment injuries. MATERIAL AND METHODS: In Finland, the Patient Insurance Center (PIC) provides financial compensation for patients who have sustained an injury in connection with medical treatment or operation. We retrospectively analyzed all claims for compensation arising from treatment of tibial fractures in children that had been received by the PIC between 1997 and 2004. The mode of treatment, complications, and permanent sequelae were assessed. We also estimated the number of avoidable treatment injuries. RESULTS AND INTERPRETATION: The PIC received 50 claims for compensation during the 8-year study period. The claims were based on the following issues: pain, incorrect diagnosis and treatment, permanent disability, extra treatment expenses, inappropriate behavior of the medical personnel, and loss of income of the parents. 35/50 claims had received compensation, of which 32 were related to the treatment and 3 to infections. The treatment injuries that had led to compensation comprised a delay in diagnosis and treatment in 15 patients, inappropriate casting in 9, inappropriate operative treatment in 5, and other causes in 3 patients. An unsatisfactory standard of treatment and missed diagnosis were the most common reasons for compensation. In restrospect, all but 1 of the 35 injuries that had led to compensation were considered to be avoidable. PMID- 19297790 TI - Introduction of total knee arthroplasty in Lithuania: Results from the first 10 years. AB - BACKGROUND AND PURPOSE: We have previously reported that the first 10 years of hip arthroplasty in Lithuania resulted in a higher cumulative revision rate than that observed in Sweden. We thus compared the corresponding results after introduaction of total knee replacement in Lithuania. METHODS: The 10-year revision rate for the first 595 primary ScanKnee arthroplasties inserted in Klaipeda, Lithuania, was compared to that for the first 1,280 ScanKnee primary arthroplasties inserted in Sweden. As in the hip replacement study, only patients with osteoarthritis (OA) were included. Primary knee arthroplasties without patellar resurfacing were included, and the endpoint was revision for any reason other than addition of a patellar component. RESULTS: We found that the cumulative revision rate was not statistically significantly different between the groups. The revision pattern was different, however, and we observed 24 isolated patellar component additions in Sweden, but none in Klaipeda. INTERPRETATION: Contrary to the results of our previous hip arthroplasty study, the cumulative revision rate after total knee arthroplasty was similar in the two groups. This suggests that compared to hip arthroplasty, the outcome of total knee arthroplasty was less dependent on surgical experience. The large difference regarding isolated patellar component additions may be explained by long-term accumulation of severe OA cases in Lithuania. To patients subject to a newly introduced surgical treatment offering great improvement in quality of life, patellofemoral pain may be a minor problem. Furthermore, patellar problems may not have seemed particularly relevant for the surgeons, considering the disability of other patients waiting to be treated. PMID- 19297791 TI - Risk factors for revision after shoulder arthroplasty: 1,825 shoulder arthroplasties from the Norwegian Arthroplasty Register. AB - BACKGROUND AND PURPOSE: Previous studies on shoulder arthroplasty have usually described small patient populations, and few articles have addressed the survival of shoulder implants. We describe the results of shoulder replacement in the Norwegian population (of 4.7 million) during a 12-year period. Trends in the use of shoulder arthroplasty during the study period were also investigated. PATIENTS AND METHODS: 1,531 hemiprostheses (HPs), 69 total shoulder replacements (Neer type TSR), and 225 reversed total shoulder replacement (reversed TSR) operations were reported to the Norwegian Arthroplasty Register between 1994 and 2005. Kaplan-Meier failure curves were drawn up for particular subgroups of patients, and revision rates were calculated using Cox regression analysis. RESULTS: The 5- and 10-year failure rates of hemiprostheses were 6% (95% CI: 5-7) and 8% (95% CI: 6-10), and for reversed total shoulder replacements they were 10% (95% CI: 5-15) and 22% (95% CI: 10-33), respectively. For hemiprostheses, the risk of revision for patients who were 70 years or older was half that of those who were younger (RR = 0.47, CI: 0.28-0.77), while the risk of revision was highest for patients with sequelae after fracture compared to those with acute fractures (RR = 3.3, CI: 1.5-7.2). No differences in prosthesis survival were found between the different hemiprosthesis brands. The main reasons for revision of hemiprostheses were pain and luxation. For reversed total prostheses, the risk of revision was less for women than for men (RR = 0.26, CI: 0.11-0.63), and the main cause of revision was aseptic loosening of the glenoid component. During the study period, the incidence of shoulder arthroplasty increased for all diagnostic groups except inflammatory arthritis, for which a decrease was seen. INTERPRETATION: We found good results in terms of 5-year prosthesis failure rate, with the use of hemiarthroplasty for patients with inflammatory arthritis, osteoarthritis, and acute fractures. Reversed total shoulder replacement was associated with a rather poor prognosis. PMID- 19297792 TI - Does fixed-angle plate osteosynthesis solve the problems of a fractured proximal humerus? A prospective series of 87 patients. AB - BACKGROUND AND PURPOSE: There is considerable controversy about the treatment of complex, displaced proximal humeral fractures. Various types of head-preserving osteosynthesis have been suggested. This prospective case series was designed to evaluate the perioperative and early postoperative complications associated with fixed-angle implants and to record outcome after bone healing. PATIENTS AND METHODS: Fractures of the proximal humerus were stabilized surgically in 87 patients (mean age 64 (16-93) years) by application of a fixed-angle plate (65 PHILOS, 22 T-LCP). There were 34 2-segment fractures, 42 3-segment fractures, and 11 4-segment fractures, including 7 dislocation fractures. Follow-up assessment after a minimum of 12 months was based on the Constant, UCLA, and DASH scores and on radiographs. RESULTS: Postoperative complications included soft tissue problems (n = 9), humeral head necrosis (n = 9), screw perforation (n = 11), secondary displacements (n = 14), and delayed fracture healing (n = 4). Treatment outcomes recorded on the various scores were very good in 60-82% of the cases. INTERPRETATION: Screw perforation of fixed-angle implants has replaced the complications of secondary displacement and implant loosening after using conventional plates. Even with the use of fixed-angle implants, fractures of the proximal humerus are associated with a high complication rate and sometimes poor outcome. PMID- 19297793 TI - Newly discovered mutations in the GALNT3 gene causing autosomal recessive hyperostosis-hyperphosphatemia syndrome. AB - BACKGROUND AND PURPOSE: Periosteal new bone formation and cortical hyperostosis often suggest an initial diagnosis of bone malignancy or osteomyelitis. In the present study, we investigated the cause of persistent bone hyperostosis in the offspring of two consanguineous parents. METHODS: Clinical assessment, imaging, and direct sequencing were used to elucidate the etiology of the condition seen in the patient. RESULTS: Radiological examination revealed periosteal reaction, diaphysitis, and cortical hyperostosis, suggesting osteomyelitis or a bone neoplasm. The clinical and radiological features were also reminiscent of hyperostosis with hyperphosphatemia (HHS), a rare autosomal recessive disease manifesting with recurrent, transient, and painful swelling of the long bones. The identification of two novel heterozygous pathogenic mutations in the GALNT3 gene confirmed a diagnosis of HHS. INTERPRETATION: Molecular analysis represents an invaluable tool in the differential diagnosis of persistent cortical hyperostosis. PMID- 19297794 TI - The effects of nail rigidity on fracture healing in rats with osteoporosis. AB - BACKGROUND AND PURPOSE: Stress shielding from rigid internal fixation may lead to refracture after removal of the osteosynthesis material. We investigated the effect of a low-rigidity (Ti-24Nb-4Zr-7.9Sn) intramedullary nail regarding stress shielding and bone healing of osteoporotic fractures in the rat. METHODS: 40 female Sprague-Dawley rats, aged 3 months, were divided into the following groups: sham-operation (SHAM) (n = 10), ovariectomized (OVX) (n = 10) and OVX fracture (n = 20). 10 SHAM rats and 10 OVX rats were killed after 12 weeks to provide biomechanical data. Ovariectomy was performed 12 weeks before fracturing both femurs in 20 rats. The left fracture was stabilized with a high-rigidity titanium alloy pin (Ti-6Al-4V; elastic modulus 110 GPa) and the right with a low rigidity (Ti-24Nb-4Zr-7.9Sn; elastic modulus 33 GPa). The bony calluses were examined by micro-CT at 6 and 12 weeks after fracture, bone volume (BV) and total volume (TV) were determined at the callus region (ROI1) and the total femur (ROI2). Subsequently, the bones were tested mechanically by a three-point bending test. RESULTS: In the low-rigidity group, TV (ROI1) increased at 6 weeks, but BV (ROI1), BV (ROI2) were similar but maximum load increased. At 12 weeks, the maximum load and also BV (ROI1, ROI2) were increased in the low-rigidity group. INTERPRETATION: The low-rigidity nail manufactured from Ti-24Nb-4Zr-7.9Sn showed better external callus formation, seemed to reduce effects of stress shielding, and reduced bone resorption better than the stiffer nail. The low-rigidity nail was strong enough to maintain alignment of the fracture in the osteoporotic rat model without delayed union. PMID- 19297795 TI - Seeking for the ideal tale pleurodesis method. PMID- 19297796 TI - The endovascular approach to abdominal aortic aneurysm repair. AB - Endovascular aneurysm repair (EVAR) affords patients the opportunity to undergo a less invasive method of repairing abdominal aortic aneurysms. The EVAR procedure results in a shorter hospital stay, less pain, and less chance of morbidity. This procedure can be performed in a variety of settings (eg, OR, interventional radiology suite, cardiac catheterization laboratory), each of which has its own benefits and challenges. Recently, hybrid ORs have been designed to perform both endovascular and open procedures. AORN has many applicable recommended practices to guide perioperative nurses in the safe provision of care during EVAR procedures. PMID- 19297797 TI - Do surgical personnel really need to double-glove? AB - In 2007, AORN's Recommended Practices Task Force revised the "Recommended practices on prevention of transmissible infections in the perioperative practice setting" to recommend that health care practitioners double-glove during invasive procedures. Previously, AORN had suggested that wearing two pairs of gloves might be indicated for some procedures. Research on the protective effects of double gloving provides compelling evidence that surgical personnel should double-glove during all surgical procedures. Statistics on unreported injuries and conversion rates of HIV and hepatitis B indicate that change is difficult; however, research also suggests that objections to double-gloving can be overcome and a practice change implemented to ensure the safety of health care workers and patients alike. PMID- 19297798 TI - [And the winner is... ]. PMID- 19297799 TI - [Distant healing: expectations, relationships and solutions]. PMID- 19297800 TI - [Relaxation practice modifies gene expression]. PMID- 19297802 TI - Changes in drug utilization for seniors without prior prescription drug insurance. AB - (1) After the passage of the Medicare Modernization Act, a majority of individuals without previous prescription drug coverage obtained coverage under Medicare Part D. (2) Among previously uninsured seniors, utilization of prescription drugs increased with the introduction of Medicare Part D. There was rapid uptake of newly marketed generics. (3) Dual eligibles had little change in prescription utilization or rates of switching between drugs after the implementation of Part D. Out-of-pocket spending was significantly reduced for this population. PMID- 19297801 TI - STrengthening the REporting of Genetic Association studies (STREGA)--an extension of the STROBE statement. AB - Making sense of rapidly evolving evidence on genetic associations is crucial to making genuine advances in human genomics and the eventual integration of this information in the practice of medicine and public health. Assessment of the strengths and weaknesses of this evidence, and hence the ability to synthesize it, has been limited by inadequate reporting of results. The STrengthening the REporting of Genetic Association studies (STREGA) initiative builds on the STrengthening the Reporting of OBservational Studies in Epidemiology (STROBE) Statement and provides additions to 12 of the 22 items on the STROBE checklist. The additions concern population stratification, genotyping errors, modelling haplotype variation, Hardy-Weinberg equilibrium, replication, selection of participants, rationale for choice of genes and variants, treatment effects in studying quantitative traits, statistical methods, relatedness, reporting of descriptive and outcome data and the volume of data issues that are important to consider in genetic association studies. The STREGA recommendations do not prescribe or dictate how a genetic association study should be designed, but seek to enhance the transparency of its reporting, regardless of choices made during design, conduct or analysis. PMID- 19297803 TI - KRAS mutations and epidermal growth factor receptor inhibitor therapy in metastatic colorectal cancer. PMID- 19297804 TI - Special report: recent developments in prostate cancer genetics and genetic testing. PMID- 19297805 TI - Special report: MEG and MSI for the purpose of presurgical localization of epileptic lesions--a challenge for technology evaluation. PMID- 19297806 TI - Special report: early intensive behavioral intervention based on applied behavior analysis among children with autism spectrum disorders. PMID- 19297807 TI - [Third Francophone Surgical Congress, June 25-28, Quebec]. PMID- 19297808 TI - [Are cost savings the best medicine for the ailing health care?]. PMID- 19297809 TI - [Good continuity of patient care will give better care results. Strong measures necessary to provide continuing contacts with physicians in Swedish health care]. PMID- 19297810 TI - [Orbital fractures in children require immediate intervention. Risk of permanent double vision]. PMID- 19297811 TI - [High-precision orbital reconstruction possible with new implants]. PMID- 19297812 TI - [Functions against the odds. Primary health care in Palestine supported by Sweden]. PMID- 19297813 TI - [Psychopathy and brain imaging--a literature review. With focus on functional magnetic resonance tomography]. PMID- 19297814 TI - [Coronary computer tomography--time for reflection. A survey of the current knowledge]. PMID- 19297815 TI - [Also heterosexual men can get urinary tract infection via anal sex]. PMID- 19297816 TI - [How is it with "lean healthcare"? Decentralized, professional care can steer us in right course]. PMID- 19297817 TI - [Remains to prove if "lean thinking" is beneficial for health care]. PMID- 19297818 TI - ["Care Choice Halland was created for local inhabitants"]. PMID- 19297819 TI - [Final reply on medical education at the Karolinska Institute: freedom of choice can not substitute basic clinical knowledge]. PMID- 19297820 TI - [Points for continuing education not necessary]. PMID- 19297821 TI - [Bone marrow biopsy not necessary for the diagnosis of polycythemia vera]. PMID- 19297822 TI - [Correct but partly misleading about a laboratory test]. PMID- 19297823 TI - [Observation studies from Stockholm during the time of cholera]. PMID- 19297824 TI - [Proposal from the Swedish National Council on Medical Ethics is no capitulation]. PMID- 19297825 TI - [Cognitive failure after surgery--a multidisciplinary challenge]. PMID- 19297826 TI - [Postoperative cognitive dysfunction can be connected to 1-year mortality. The role of anesthetics is not yet clear]. PMID- 19297827 TI - [Anemia a common diagnosis in elderly. How low can the Hb value be?]. PMID- 19297828 TI - [Pain. Multidisciplinary attitude decisive]. PMID- 19297829 TI - [Pain analysis--basis for management of long-term pain]. PMID- 19297830 TI - [Pain analysis creates physician-patient confidence]. PMID- 19297831 TI - [Quality of life measurement important in long-term pain]. PMID- 19297832 TI - [Sexual dysfunction common in long-term pain]. PMID- 19297833 TI - [Neuropathic pain can be handled in ten minutes]. PMID- 19297834 TI - [Neurostimulation is effective in severe angina pectoris]. PMID- 19297835 TI - [Should breast feeding mothers dispose of their milk after anesthesia?]. PMID- 19297836 TI - [Inadequate basic knowledge among students from the Karolinska Institute?]. PMID- 19297837 TI - [Memory recall as treatment lacks scientific support]. PMID- 19297838 TI - Solar panels make really good cents. AB - Bobby Hancock, senior director of facility management for the Bloorview Kids Rehab facility in Toronto, describes how features such as a 37 kW penthouse roof solar array, thermal glazed windows, rainwater harvesting, and air handling units with variable speed drives and heat recovery wheels, contribute to the "green" credentials of Canada's largest children's rehabilitation centre. PMID- 19297839 TI - Retrofits provide re-think opportunities. AB - There are many factors to be considered during a retrofit, not least of which should be the building control system. But as Sean McGowan discovered, this is often the one element not given due consideration, resulting in poor performance, higher running costs, and elevated greenhouse emissions, despite the system's aims. PMID- 19297840 TI - Building relationships for better maintenance. AB - Effective management of building maintenance is a vital ingredient in ensuring a high quality built environment for any building's occupiers. However, maintenance is not high on the list of priorities for most organisations. Communication between top management personnel looking at maintenance issues from a strategic standpoint and maintenance staff considering them at an operational level is often not as good as it should be. When planning maintenance activities maintenance personnel often draw too heavily on their technical experience and expertise without taking sufficient account of wider organisational objectives or consulting effectively with top management. Senior managers also regularly complain about lack of managerial input from maintenance departments. Such barriers contribute to communication difficulties between top management at a strategic level and maintenance personnel at an operational level. Identifying where the key differences lie in senior managers' and maintenance personnel's viewpoints on maintenance strategy can prove invaluable in achieving some convergence of opinions and optimising the efficiency of the overall building maintenance process. PMID- 19297841 TI - Capitalising in the long term. AB - Bernd Rechel, Stephen Wright, Nigel Edwards, Barrie Dowdeswell and Martin McKee report on an international study on improving the effectiveness of health capital investment which analyses innovative European capital projects and examines the key issues that arise. The study, by the European Observatory on Health Systems and Policies and the European Health Property Network/European Centre for Health Assets & Architecture, suggests that hospital design that takes into account facility management costs results in significant efficiency gains. PMID- 19297842 TI - Manslaughter Act may catch out the unwary. AB - The key areas that NHS Trusts and other organisations running healthcare facilities need to consider, and the risk audit processes recommended, to minimise their risk of prosecution under the Corporate Manslaughter & Homicide Act should fatalities occur on their premises, were examined by Gary Rogers, a chartered surveyor at Faithful + Gould, at the Healthcare Facilities Consortium's recent 2008 Annual Conference. PMID- 19297843 TI - Hospital parking--the big squeeze. AB - With the British public far more likely than they were 10 to 15 years ago to use a car even for a short journey, it seems inevitable that it will gradually become more and more difficult to find a hospital car park space. Jonathan Baillie discusses the key issues for health estate, facilities and transport managers dealing with car parking and traffic management on often congested, and increasingly constricted, sites. PMID- 19297844 TI - Building with less waste should be goal. PMID- 19297845 TI - Visual contrast spotlighted. AB - Following the publication of the new British Standard for the measurement of light reflectance values (LRVs), Keith Oakes, technical director for commercial and industrial flooring manufacturer Gradus, examines the importance of visual contrast in healthcare building design. PMID- 19297846 TI - Introducing BREEAM for Healthcare. AB - Andrew Welfle, sustainability consultant at Faber Maunsell, discusses the recently introduced BREEAM for Healthcare assessment system for health sector buildings, and highlights the key elements of a building's design that contribute to a positive rating. PMID- 19297847 TI - Welsh Trust sees energy costs reduce. AB - Working closely with Honeywell Building Solutions, a division of Honeywell International, under two long-term Energy Performance Contracts, should bring Gwent Healthcare NHS Trust in South Wales guaranteed annual savings of over pounds 1.1 million in energy costs over at least the next 15 years, as the Trust's estates manager, Wayne Churches, explained to delegates at the Healthcare Facilities Consortium (HFC) 2008 Annual Conference. PMID- 19297848 TI - [The value of the fiber laryngoscope in the diagnosis of congenital laryngeal stridor]. AB - OBJECTIVE: To explore the value of fiber laryngoscope in the diagnosis of congenital laryngeal stridor (CLS). METHOD: The clinical data of One hundred and thirty children who had been examen by laryngofiberscopy and had laryngeal stridor were analyzed retrospectively. RESULT: We found 73 children having CLS, 2 children having laryngeal web, 5 children having congenital epiglottic cyst, 20 children having mass and cyst of tongue root, 1 child having laryngocele, 3 children having laryngeal papilloma, unilateral or bilateral cricoarytenoid joint of 9 children having poor activities, 17 children were normal. CONCLUSION: CLS is misdiagnosed easily. Laryngeal strio caused by other diseases soule be excluded before CLS is diagnosed. Our proposal is that fiber laryngoscope should be used as a routine method in the diagnosis of CLS. CT and MRI could also be used if it is necessary. PMID- 19297849 TI - [The expressions and clinical significance of tumor suppressor gene CX26 in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the expressions of tumor suppressor gene CX26 mRNA and coding protein in laryngeal squamous cell carcinoma, and to explore the relationship between CX26 gene and the biological behaviors of laryngeal squamous cell carcinoma for understanding the tumorigenicity and development of laryngeal squamous cell carcinoma. METHOD: Laryngeal carcinoma tissues (studying group), which takeda from the center of tumors and laryngeal normal tissues (control group) takeda at the place of 1.0 cm out of the edge of the tumors, were took from 38 patients with laryngeal squamous cell carcinoma while they were in operation. Semi-quantitative reverse transcriptase polymerase chain reaction (RT PCR) was used to analyze the expression level of CX26 mRNA, and immunohistochemical staining (frozen section) was used to detect the expression of CX26 protein in laryngeal carcinoma tissues and laryngeal normal tissues of 38 cases, respectively. RESULT: mRNA of CX26 gene was all positively expressed in laryngeal carcinoma tissues and laryngeal normal tissues of 38 cases by RT-PCR. However, CX26 mRNA was obviously down-regulated in laryngeal carcinoma tissues than that in laryngeal normal tissues (P < 0.05). Immunohistochemical staining showed CX26 protein was strong-positively expressed in laryngeal normal tissues in 34 cases (89.5%), while it was positively expressed in laryngeal carcinoma tissues in 18 cases (47.4%), and with the location alteration of CX26 protein in laryngeal carcinoma cells. There was significant difference between the expression rate of CX26 protein in laryngeal carcinoma tissues and in laryngeal normal tissues (P < 0.05). Meanwhile, the expression level of CX26 mRNA and the positive-expressed rate of CX26 protein of the laryngeal carcinoma tissues in the advanced stage patients group (III stage and IV stage) were significantly lower than these in the early stage patients group (I and II) (P < 0.05), and it was significantly lower in those who have a cervical lymph node metastasis than those without metastasis. (P < 0.05). Moreover, the expression level of CX26 mRNA and the positive-expressed rate of CX26 protein reduced along with the reduction of pathological differentiation, and there was significant difference among the well differentiated group, moderately-differentiated group and poorly-differentiated group (P < 0.05). CONCLUSION: CX26 gene may play an important role in the pathogenesis and development of laryngeal carcinoma and may be related to its prognosis. PMID- 19297850 TI - [The expression and significance of integrin beta1 and focal adhesion kinase and its clinical value in laryngeal carcinoma]. AB - OBJECTIVE: To study the expressions and clinical significance of integrin beta1 and focal adhesion kinase (FAK) in laryngeal carcinoma (LSCC). METHOD: The mRNA and protein levels of integrin beta1 and focal adhesion kinase and the surrounding tissue of laryngeal carcinoma in 48 specimens and 20 specimens of vocal cord polyp were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry SABC method. RESULT: The mRNA levels and positive rates of integrin beta1 and FAK protein were significantly higher in laryngeal carcinoma than that in the surrounding tissue and vocal cord polyp (P < 0.05). The expression levels of integrin beta1 and FAK were significantly higher in the group with cervical lymph node metastasis than those without (P < 0.05), and they were significantly higher in the tissue of stage of T3 and T4 than those of T1 and T2 (P < 0.05). But pathological grades was not significantly related with the expression levels of integrin beta1 or FAK (P > 0.05). CONCLUSION: The expression levels of integrin beta1 and FAK were increased in LSCC, and they may contribute significantly to invasion and metastasis of LSCC. PMID- 19297851 TI - [Expression and significance of PD-L1 in laryngocarcinoma]. AB - OBJECTIVE: To study the expression of ligand 1 of programmed death-1 (PD-L1) in human laryngocarcinoma tissues and peritumoral tissue and its potential significance to clinic. METHOD: Immunohistochemistry and Western-blotting were performed to assess the expression of PD-L1 at protein level and the differences in 31 cases of laryngocarcinoma tissue and 24 cases of paraneoplastic tissue. RESULT: Expression of PD-L1 was significantly higher in poorly and moderately differentiated larynx squamous carcinoma than that in well differentiated larynx squamous carcinoma. The expression of PD-L1 in the former was mainly in blood vessels while rarely in cancer cells, and the blood vessel of the latter displayed weakly positive while partial cancer cells showed positive; and the expression of PD-L1 was significantly correlated with lymphatic metastasis. Expression of PD-L1 in paraneoplastic tissues was significantly lower than that in cancer tissues but positively correlated to the expression level in the cancer tissues. CONCLUSION: PD-L1 may play a part in the development and/or metastasis of laryngocarcinoma, particularly in the bad differentiated; And it maybe a new target of immunotherapy for laryngocarcinoma, especially for the poorly and moderately differentiated. PMID- 19297852 TI - [The evaluation of quality of life for obstructive sleep apnea hypopnea syndrome in children]. AB - OBJECTIVE: One of the objective of the study was to evaluate the reliability and validity of 18 items survey (OSA-18) which was made for disease specific quality of life in children with obstructive sleep apnea. The other was to explore whether OSA-18 could be used as a well appreciation method on OSAHS in children's quality of life before and after surgery. METHOD: First, one hundred and twenty two children's parents were interviewed with this survey scales and the survey scales was assessed by split-half reliability, retest reliability, internal consistency, construct validity and content validity. Second,OSA-18 was used to evaluate the QOL of 122 children within 4 weeks before PSG and in 6 to 12 months after operation. RESULT: OSA-18 has a satisfactory internal consistency. Global Cronbach's alpha coefficient were 0.939. All domains and items Cronbach's alpha coefficients > 0.6; also has a well retest reliability, coefficient correlation of pearson equal 0. 619. Construct validity and content validity were satisfactory. The impact of pediatric OSAHS on QOL was moderate and severe in 77.05% patients pre-operation. However, it was only 28.69% after surgery. OSA-18 total scores, every domain and item score were significant decrease. CONCLUSION: OSA-18 QOL survey had better reliability and validity, and was applicable for life quality assessment in OSAHS children. OSA-18 QOL survey could help clinical diagnosis of OSAHS in children and give quantitative evaluation for therapeutic measure. PMID- 19297853 TI - [Observe the curative effect of n-Butyl cyanoacrylate injection in treating laryngopharynx hemangioma]. AB - OBJECTIVE: To evaluate the clinical effectiveness of injection n-Butyl cyanoacrylate (NBCA) in treating laryngopharynx hemangioma. METHOD: Thirty cases of laryngopharynx hemangioma who received NBCA injection were obtained from our department 1998-2007. Twenty-five cases had tracheotomy under General anesthesia, and NBCA was injected into hemangioma by direct laryngoscopy. NBCA was mixed with iodide, and the concentration was 25.00% to 33.33%, NBCA dosage was 0.5 ml to 2.0 ml. There were 5 cases whose hemangioma confined to oropharynx didn't have tracheotomy, and they were injected straightly with the speculum oris. We observed the shedding of hemangioma in different time. RESULT: Hemangioma with diameter less than 1 cm, shedded in about 1 month. However, hemangioma with diameter more than 4 cm, shedded in a period more than 3.5 month. No recurrence was observed in the follow up of 3 months to seven years. CONCLUSION: Avoiding dissection of neck and repetitious operation, NBCA injection could be a safe, simple and effective therapy for laryngopharynx hemangioma. PMID- 19297854 TI - [Comparing between CAUP and UPPP in treatment of OSAHS]. AB - OBJECTIVE: To evaluate the prospective effect of CAUP and applying midline partial glossectomy with UPPP in the treatment of OSAHS. METHOD: One hundred OSAHS patients were distributed into two groups averagely. The patients of the two groups had CAUP and midline partial glossectomy respectively. All patients were followed-up 6 months and 12 months after operation and carried out using PSG every time. RESULT: Therapeutic effect was evaluated by standard of Hangzhou (2002). In general, in 6 months, curative effect was excellent in 41 patients of the group of CAUP, good in 8 patients with effect, bed effect in 1 patients; in 12 months, it was excellent in 28 patients, good in 13 patients, bed in 9 patients. In 6 months, the curative effect the other group were excellent in 44 patients, good in 5 patients, bed in 1 patients; In 12 months, it was excellent in 43 patients , good in 6 patients, bed in 1 patients. In 6 month, the results showed that the effect of the two methods was similar (P > 0.05), but in 12 months the effect of group of CAUP was better than the other obviously (P < 0.01). CONCLUSION: This operation could enlarge the narrow area of palatopharyngeal cavity effectively and decrease postoperative complications. It's a safe effective and acceptable surgical procedure. PMID- 19297855 TI - [Relationship between the polysomnography and body mass index in children]. AB - OBJECTIVE: To study the effect of the obstructive sleep apnea hypopnea syndrome on children's growth and to discuss operative standards. METHOD: Five hundred and eighty-two children who were 2.5 to 10 years old were monitored with polysomnography(PSG). The index included apnea and hypopnea index (AHI) and computed body mass index (BMI) which was measured with heights and weights. RESULT: Comparing with the normal children in the same age period and the same sex, boys' BMI had distinguished differences (P < 0.01) when AHI > or =2 and girls BMI had distinguished differences (P < 0.01) when AHI > or =1. CONCLUSION: We Suggests to define the diagnosis and operative standards of pediatric obstructive sleep apnea hypopnea syndrome at AHI > or =2. PMID- 19297856 TI - [Plasma radiofrequency ablation in treatment of recurrence of adenoidal hypertrophy]. AB - OBJECTIVE: To investigate the long-term outcome and clinical value of utilizing plasma radio frequency ablation under indirect laryngoscope in treatment of recurrence of adenoidal hypertrophy. METHOD: Fifty patients with recurrence of adenoidal hypertrophy were subjected to the operation designed by our group. RESULT: The period of follow-up was more then 2 years. All patients were free of snore postoperatively; conductive hearing loss was improved as well. CONCLUSION: Operation with plasma radiofrequency ablation had advantages of direct and clear view of surgical area, accurate remove of adenoids, non-bleeding, avoiding damage of normal nasopharyngeal structure structures, minimal invasion, high safety, and little complication, in treatment of recurrence of adenoidal hypertrophy. PMID- 19297857 TI - [The experimental investigation of laryngeal cancer growth in vive by recombinant lentivirus of small interference RNA (siRNA) targeting MMP-9]. AB - OBJECTIVE: To investigate the inhibitive role of lentivirus mediated MMP-9 gene silence in the growth of laryngeal cancer xenografts. METHOD: The nude mouse model of laryngeal squamous carcinoma was established and RNA interference (RNAi) technic was used. Recombinant MMP-9-RNAi-Lentivirus was intratumoral injected while the empty vector lentivirus was taken as control. After the treatment, tumor inhibitive effect was observed and western-blot was used to test the expression of MMP-9 in the xenografts. Finally, the PCNA expression in xenografts were examined with immunohistochemistry to evaluate proliferation change of the Hep-2 cells. RESULT: The average tumor weight was (1.484 +/- 0.391) g in group treated with MMP-9-RNAi-Lentivirus which was significantly lower than that (2.618 +/- 0.465) g in the control group (P < 0.05). The average tumor volume was (1.177 +/- 0.270) cm3 in group treated with MMP-9-RNAi-Lentivirus which was significantly lower than that (2.034 +/- 0.366) cm3 in the control group (P < 0.05) and the tumor inhibitive rate was 43.32%. Western-blot showed there were 7 cases of xenografts with MMP-9 protein negative expression and the other 3 cases were tested with weak positive expression in the treated group xenografts. Whereas all the 10 cases of xenografts were tested with MMP-9 positive expression in the control group. Immunohistochemistry showed PCNA index in treated group was (55.41 +/- 8.77)% which was obviously lower than that (77.04 +/- 6.91)% in control group (P < 0.01). CONCLUSION: The growth and proliferation of laryngeal cancer could be inhibited by recombinant MMP-9-RNAi-Lentivirus. PMID- 19297858 TI - [The inhibitory role of zhongjiefeng extracts, on apoptosis and telomerase activity of nasopharyngeal carcinoma cell line xenograft in nude mice]. AB - OBJECTIVE: This study was designed to evaluate the effect of zhongjiefeng extracts on telomerase activity and apoptosis of implanted human nasopharyngeal carcinoma cell lines in nude mice. METHOD: Nude mice with implanted human nasopharyngeal carcinoma cell lines CNE1 and CNE2 were randomly divided into three groups, which were the group of zhong-jiefeng, normal control group, and CTX therapy group. The weight of nude mice and the volume of tumor were regularly measured. After a course of treatment (30 d), tumor tissues were obtained with autopsy, and then they were weighed to calculate the inhibitive rate of the growth of tumor. The changes of ultra micro-structure of NPC cells were observed by transmission electron microscope. Expressions of Bcl-2 and Bax of xenografts were investigated by immunohistochemistry. The apoptosis of CNE1 and CNE2 cells was observed using TUNEL and FACS with PI-staining. The telomerase activity was observed by TRAP-ELISA method. RESULT: The zhongjiefeng extracts significantly inhibit the growth of tumor compared to normal control group. The inhibitory rate were 40.8% (P < 0.01) and 46.8% (P < 0.01), respectively. The expression of Bcl-2 was lower (P < 0.01) and the expression of Bax was higher (P < 0.01) in the zhongjiefeng group than that in normal control group. Electron microscopy indicated the typical apoptosis of tumor cells, such as marginal nuclei, chromatin condensation and nuclei fragmentation, and apoptotic bodies. As result of the flow cytometry showed, the exhibition rate of apoptosis of cell in the group treated with zhongjiefeng was higher than that of normal control group (P < 0.01) in G0/G1 phase. Zhongjiefeng arrested cells in G0/G1 phase of the cell cycle. The telomerase activity of zhongjiefeng and CTX group was lower than that of the normal group (P < 0.01). CONCLUSION: Zhongjie feng suppressed the growth of tumor in vivo. The anticancer effects of zhongjiefeng were associated with the induction of apoptosis and partly with the suppression of telomerase activity. PMID- 19297859 TI - [Analysis of different distribution of causes of hoarseness between the Han and the Uighur]. AB - OBJECTIVE: To evaluate clinical characteristics of distribution of causes of hoarseness in the Han and the Uighur. METHOD: The date of 933 cases of hoarseness in different ages, which included 654 cases of the Han and 279 cases of the Uighur, were analyzed with laryngofiberoscope. They were divided into different age groups and were compared. RESULT: The common causes of hoarseness of the Han, which occupied 90.1% of its all cases, were polyp of vocal cord (31.8%), chronic laryngitis (24.9%), vocal nodule (12.2%), carcinoma of larynx (11.2%), paralysis of vocal cord (9.9%); While that of the Uighur, which occupy 96.4% of its all cases, were chronic laryngitis (27.2%), paralysis of vocal cord (21.1%), polyp of vocal cord (19.4%), carcinoma of larynx (12.9%), laryngeal papillomatosis (7.9%), vocal nodule (7.9%), there was significant difference between them (chi2=73.19, P < 0.01) and significant difference between them in distribution of polyp of vocal cord,paralysis of vocal cord and laryngeal papillomatosis (P < 0.01). There also significant difference among 2 to 20 years old group, 21 to 40 years old group and 41 to 60 years old group. However 61 to 85 years old group was excepted. CONCLUSION: There was difference in distribution of causes of hoarseness between the Han and the Uighur, which was also different in different ages of them. The main causes of hoarseness in the Han were chronic laryngitis and vocal nodule in children and teenagers,polyp of vocal cord and chronic laryngitis in youth and adult, while in the Uighur, they were chronic laryngitis and laryngeal papillomatosis in children and teenagers, chronic laryngitis, polyp of vocal cord and paralysis of vocal cord in youth and adult; In the aged, there was no Statistical difference between the two nations, and the major cause was carcinoma of larynx and paralysis of vocal cord. PMID- 19297860 TI - Overcapacity protocols. PMID- 19297861 TI - ISMP Medication Safety Alert! Falsely elevated glucose readings from maltose containing drugs. PMID- 19297862 TI - Making a difference ... one hand wash at a time. Interview by Sue Ridewood,. PMID- 19297863 TI - Being present. PMID- 19297864 TI - Bioethics, biopolitics and medical regulation: learning from the Nazi doctor experience. AB - The phenomenon of abuse committed by medical practitioners during the Nazi era generated the creation of a series of important human rights instruments. Its evolution from sterilisation of persons with intellectual disabilities and euthanasia of those with mental illnesses constituted a dreadful fusing of legal and medical initiatives. Together these made possible the dehumanising rhetoric and then actions against Jews, the creation of the death camps by doctors, and the commission of human experiment atrocities by medical practitioners at the camps. There remains much that we must remember and learn if we are to formulate effective checks and balances to reduce the risk or repetition of such conduct. Numbered among these is a preparedness to name "evil" when it occurs so as to facilitate distinctions being drawn between conduct that is professionally acceptable and that which is repugnant and inconsistent with all principles of ethical conduct by medical practitioners. PMID- 19297865 TI - Hospital orders for offenders with mental illnesses: an appropriate diversionary option? AB - Hospital orders provide an option for sentencing judges to divert offenders with mental illnesses away from the criminal justice system and into the civil commitment system for treatment purposes. Such an option exists in Victoria, Tasmania and at a Commonwealth level. However, hospital orders have been rarely used. This column explores some of the practical difficulties associated with such orders, with the lack of resources for proper treatment and services an ever present issue. It outlines the recent changes to the Victorian scheme and argues that if properly resourced, hospital orders can provide a means for adequately addressing the treatment needs of offenders with mental illnesses. PMID- 19297866 TI - Regulation of health practitioners by trade practices and fair trading legislation. AB - In a variety of situations, particularly those characterised by commerciality, corporate structures and unregistered practitioners, there are major limitations to traditional regulation by health boards and councils, as well as hearings by external tribunals. Part of the difficulty lies with the ability of external bodies to award compensation to complainants/notifiers proved to have suffered adverse consequences from proven unprofessional conduct. This column advances suggestions for reform of the powers of external tribunals to redress this deficit. It also reviews the benefits of an associated form of regulation by the Australian Competition and Consumer Commission and offices of fair trading to enable consumer protection. It reviews recent decisions in the Federal Court of Australia and the Supreme Court of Victoria in such matters as well as recommendations in 2008 by the Victorian Health Services Commissioner. PMID- 19297867 TI - Naked regulators: moral pluralism, deliberative democracy and authoritative regulation of human embryonic stem cell (hESC) research. AB - Bioethical issues pose challenges for pluralist, democratic societies due to the need to arbitrate between incompatible views over fundamental beliefs. The legitimacy of public policy is increasingly seen to depend on taking public consultation seriously, and subsequently regulating contested activities such as therapeutic cloning and hESC research. In December 2006, the Australian Federal Parliament lifted the ban on therapeutic cloning, following recommendations of the Legislation Review Committee (Lockhart Committee), which recently reported on its approach and methods in this journal. This column analyses recent accounts of democratic deliberative processes, authoritative regulation and the committee's own account. Authoritative regulation turns out to be largely an appeasement strategy, directed towards the losers of the contest, in this case the opponents of therapeutic cloning and hESC research. This is because regulation fails to minimise harm as perceived by the losers, and fails to meaningfully limit what it is the winners wish to do. Moreover, regulation adds an unnecessary layer of red tape to the work of the winners. Committees of inquiry in bioethical matters should be more open about their processes and their normative recommendations, at the risk of eroding trust in parts of their processes. PMID- 19297868 TI - New Australian federal organ and tissue donation legislation: enhanced transplant services but no "opt-out". AB - Seriously ill people who could achieve many years of high-quality life with organ donation continue to die on waiting lists due to the scarcity of donated organs. Recent advances that suggest donor organs can be coated with host stem cells to reduce or remove the need for long-term recipient immunosupressive medication highlight the importance of encouraging such donation. A wide variation in organ donor rates in developed nations suggests this is one issue in which the right regulation can make a difference. Australia has now passed federal legislation on the topic. This column considers whether that legislation is headed in the right direction, PMID- 19297869 TI - The impact of the Cartwright Report upon the regulation, discipline and accountability of medical practitioners in New Zealand. AB - The Cartwright Report instigated a profound change in thinking about patient doctor relationships and the need for public involvement in the processes by which doctors are censored. It was also the key catalyst to legislative reforms designed to ensure the accountability of practitioners to their patients. This article considers the effect of the Cartwright Report on the incidence of disciplinary hearings against medical practitioners in New Zealand. Perhaps contrary to expectations, the statistics show a pronounced decline in disciplinary hearings. The authors argue that this should not necessarily be considered an adverse development, and that these statistics in fact reflect the working of multi-layered, more constructive and open processes for regulating doctors and holding them accountable. PMID- 19297870 TI - Turning a blind eye: physical standards for surgeons. AB - Requiring professionals in high-risk industries to regularly meet minimal physical standards is a well-accepted principle. Pilots in both civil and military aviation must meet rigorous physical standards, including visual acuity, colour and peripheral vision. Australian medical practitioners have a general obligation to notify their medical registration board if they have suffered an illness which might affect their physical or mental capacity to practise medicine. However, no specific minimal physical standards are defined. It would be possible for a surgeon to be ineligible for an unrestricted motor vehicle driver's licence yet continue to perform surgical procedures. The visual requirements for a surgeon differ from those of a driver, the surgeon requiring good acuity for fine detail at close range, good depth perception as well as good colour vision. The driver needs good peripheral vision and adequate visual acuity at longer distances. However, a good yardstick for any doctor intending to perform surgical procedures would be that he or she should at least meet the Australian standard for an unrestricted driver's licence. That requirement could easily be incorporated into the annual declaration which each doctor must make when seeking renewal of her or his medical registration. PMID- 19297871 TI - Under the knife: an analysis of the Medical Council of New Zealand's Statement on Cosmetic Procedures. AB - The lack of regulation in the field of appearance medicine has long been a cause for concern in New Zealand. As an industry, it has fallen outside the protective ambit of the regulatory framework that governs all other areas of medicine. This article examines the Medical Council of New Zealand's attempt to address some of the concerns that have existed by producing a Statement on Cosmetic Procedures. The author concludes that this statement goes a long way towards better ensuring consumer safety and wellbeing in this area. It also offers valuable guidance to practitioners in the areas of advertising and promotion, obtaining consent and providing care. The author does, however, propose and discuss several possible changes that could be made to improve the statement that has been produced. PMID- 19297872 TI - Legislative intervention in Queensland to restrict access to solariums and cosmetic procedures by children and young persons. AB - Breaking new ground, Queensland has enacted laws restricting access to cosmetic surgery by those under 18 years of age. Legislation in other Australian jurisdictions is narrower in scope, focusing on niche areas such as solarium use, tattoos and body piercing. Even in those niche areas there are inconsistencies of approach and now the unique Queensland cosmetic surgery restrictions further raise the prospects of "medical tourism" and highlight the difficulties of differing legislation throughout Australia. All implementations, however, face the same challenge: to balance protection of vulnerable children, respect for a young person's autonomy and due regard to parental consent. PMID- 19297873 TI - Striving for quality use of medicines: how effective is Australia's ban on direct to-consumer prescription medicine advertising? AB - The potential for both positive and negative effects arising from direct-to consumer advertising of prescription medicines challenges health policymakers to develop regulatory schemes which selectively capture the positive aspects of the practice. Australia has dealt with this quandary by banning the practice, while New Zealand and the United States permit it. However, in recent times pharmaceutical companies have been increasingly successful in introducing promotional materials into the Australian market. This article demonstrates that the Australian ban is consistent with striving for the major policy goal of quality use of medicines, thus providing the basis for arguing that solutions to strengthen the ban against the identified threats ought to be implemented. Quality use of medicines can be most effectively achieved via the combined effect of the strengthened ban and the mimicking of the limited positive aspects of direct-to-consumer advertising by government provision of non-promotional information to consumers. PMID- 19297874 TI - The law(s) of the rings: boxing and the law. AB - To threaten harm is to assault and to realise that threat is to batter. To do so intentionally for the purpose of producing injury amounts to causing harm with intent and one cannot consent to be the victim of such violence. Despite these clearly enunciated legal principles, such conduct is routinely practised in the name of sport. Boxing is widely accepted as a highly paid professional sporting activity in which the ultimate goal is to inflict a concussive head injury upon an opponent or at least cause sufficient damage to render an opponent incapable of further self-defence. Spectators pay to watch the anticipated systematic abuse of one human being by another in much the same way they delighted in gladiators who were forced to fight for the pleasure of others. This article reviews these concepts and challenges the legal ethics of authorised violence associated with these activities undertaken in the name of sport. PMID- 19297875 TI - Scaring us all to death: the need for responsible legal scholarship on post mortem organ donation. AB - This article considers the legal, medical and policy issues arising from post mortem organ donation. It explains the basis of relevant law, and examines the diagnosis of death and the ethics of medical aspects of post-mortem donation. While the law in this area may well be imperfect, it provides an appropriate and ethical framework within which health care professionals can function. The current medico-legal framework protects and preserves the public interest, such that the broader society can be confident that the dead donor rule is observed irrespective of the way that death is diagnosed. This article also acknowledges the human fear of death and calls for responsible scholarship in this area. PMID- 19297876 TI - The Joint Commission to include patient satisfaction data on quality check. PMID- 19297877 TI - Approved: "unlinking" accreditation decisions in tailored surveys. PMID- 19297879 TI - Accepted: home care changes for home medical equipment services. PMID- 19297878 TI - Update: the 2009 decision process. PMID- 19297880 TI - Correction: 2009 disease-specific care certification manual and update. PMID- 19297881 TI - Update: accreditation and certification deposit fee information. PMID- 19297882 TI - Longitudinal examination of physical and relational aggression as precursors to later problem behaviors in adolescents. AB - Research has addressed the consequences of being a victim of physical and relational aggression but less so the consequences of being an aggressor during adolescence. Consequently, relatively little is known about the extent to which aggression in early adolescence increases the risk of later aggression and other psychosocial problems. This study involves a representative sample of seventh- and ninth-grade students from Washington State (N = 1,942). Students were surveyed on recruitment and then again 1 and 2 years later to learn about ongoing behavior problems, substance use, depression, and self-harm behaviors. Surveys also included measures of several hypothesized promotive factors: attachment to family, school commitment, and academic achievement. Findings suggest that being physically and/or relationally aggressive in grades 7 to 9 increases the risk of aggression and possibly other problem behaviors after accounting for age, gender, race, and a prior measure of each outcome. Independent promotive effects were observed in most analyses, although family attachment appeared a less robust predictor overall. Implications for prevention include acting on the behavior itself and enhancing promotive influences to lessen the risk of agression and other related problems. PMID- 19297883 TI - Is psychological aggression as detrimental as physical aggression? The independent effects of psychological aggression on depression and anxiety symptoms. AB - The differential effects of psychological and physical victimization on depression and anxiety symptoms were examined via APIM and growth curve modeling techniques in a sample of newlyweds (N = 103 couples) assessed four times over the first 3 years of marriage. On average, husbands and wives reported moderate levels of psychological aggression, and there were no sex differences in prevalence rates or mean levels. Changes in psychological victimization were associated with changes in depression and anxiety symptoms, even after controlling for the effects of physical victimization. This study demonstrates the severe impact of psychological aggression on its victims and expands on previous studies of battering samples to demonstrate that psychological victimization may be more damaging than physical victimization in nonbattering, community couples. PMID- 19297884 TI - Defining appropriate stages of change for intimate partner violence survivors. AB - Recent research suggests that the transtheoretical model of behavior change is a promising approach for interventions addressing women's experiences of intimate partner violence. This study explores the distribution of abused women across the stages of change for (a) staying safe from intimate partner violence and (b) leaving an abusive relationship. It explores the relationship between stage assignment and other indicators of a woman's stage (i.e., safety behaviors and desire for services). Quantitative surveys were conducted with 96 low-income, urban abused women recruited from six health care clinics. The findings call into question the appropriateness of using a staging algorithm that uses one "global" question about keeping safe and suggest that staging questions focused on a single action stage (e.g., leaving) are also problematic. In conclusion, additional work remains to be done to develop and validate quantitative measures of stages of change for survivors of intimate partner violence and to design, implement, and evaluated stage-based, tailored intimate partner violence interventions. PMID- 19297885 TI - Relationships between mobbing at work and MMPI-2 personality profile, posttraumatic stress symptoms, and suicidal ideation and behavior. AB - This study investigates the relationships between the experience of mobbing at work and personality traits and symptom patterns as assessed by means of the revised version of the Minnesota Multiphasic Personality Inventory (MMPI-2). Participants were 107 workers who had contacted mental health services because they perceived themselves as victims of mobbing. In line with previous research, the results showed that the MMPI-2 mean profile was characterized by a neurotic component as evidenced by elevations of Scales 1, 2, and 3 and a paranoid component as indicated by elevation of Scale 6. Contrary to previous research, a pattern of positive and significant correlations was found between the frequency of exposure to mobbing behaviors and the MMPI-2 clinical, supplementary, and content scales, including the posttraumatic stress scale. Only about half the participants showed a severity of posttraumatic stress symptoms indicative of a posttraumatic stress disorder. The frequency of exposure to mobbing predicted suicidal ideation and behavior, with depression only partially mediating this relationship. PMID- 19297886 TI - Exploring the relationship between childhood maltreatment and intimate partner abuse: gender differences in the mediating role of emotion dysregulation. AB - Despite evidence that childhood maltreatment is associated with increased risk for intimate partner abuse perpetration, the mechanisms underlying this relationship remain unclear. Given literature suggesting that violent behaviors may serve an emotion regulating function, this study examined the mediating role of emotion dysregulation in the relationship between childhood maltreatment and intimate partner abuse perpetration among 341 male and female undergraduates. However, given evidence of gender differences in the underlying mechanisms of intimate partner abuse, emotion dysregulation was expected to be more relevant to the perpetration of partner abuse among men. Consistent with hypotheses, emotion dysregulation mediated the relationship between childhood maltreatment and intimate partner abuse among men; conversely, emotion dysregulation was not associated with partner abuse among women. PMID- 19297887 TI - Intimate partner violence among hispanic men and women: the role of drinking, neighborhood disorder, and acculturation-related factors. AB - Using secondary data analysis, this study assessed the contribution of drinking, neighborhood disorder, and acculturation-related factors to past-year intimate partner violence (IPV) risk among a national sample of married or cohabiting Hispanic men (n = 1,148) and women (n = 1,399) who participated in the 2000 National Household Survey on Drug Abuse. Drinking measures were past-year alcohol abuse, past-month binge drinking, and number of past-year drinking days. Neighborhood disorder was measured by perceived level of neighborhood problems. Acculturation-related factors were nativity and survey interview language preference (Spanish vs. English). Similar proportions of men and women reported IPV perpetration (6.1% vs. 6.5%) and IPV victimization (8.8% vs. 7.8%). Logistic regression results indicated that for men, neighborhood disorder was associated with IPV perpetration (odds ratio [OR] = 1.55) and victimization (OR = 1.36). For women, neighborhood disorder (OR = 1.34) and their alcohol abuse (OR = 10.26) were associated with IPV victimization, but not IPV perpetration. Acculturation related factors were not associated with IPV perpetration or victimization for men or women. The findings suggest that IPV prevention efforts should address deleterious neighborhood conditions in addition to individual-level factors that place couples at risk for IPV. PMID- 19297888 TI - Sexual assault in men: a population-based study of Virginia. AB - Male sexual assault is a serious public health issue that is mostly underreported and unrecognized. A population-based study was conducted on 705 adult men in Virginia. The study found a lifetime prevalence of 12.9% among men, with 94% assaulted for the first time before the age of 18. Regression analysis showed that male victims of sexual violence were 3.4 times more likely to be depressed and 2.4 times more likely to ideate suicide. However, only 15.4% sought counseling. This study confirms that male sexual assault is a prevalent problem. It is also evident that victimized men were more likely to be depressed and ideate suicide and yet did not seek health services. Efforts should be made to reach men with a history of sexual assault. PMID- 19297889 TI - Intimate partner violence and community service needs among pregnant and postpartum Latina women. AB - Health care providers are advised to refer abused women to needed community services. However, little is known about abused women's perceived need for services, particularly among Latina women. We examined the relationship between intimate partner violence (IPV) and perceived needs for legal, social, and job services among a prospective cohort of 210 pregnant Latinas. IPV was associated with needing social and legal services at most time points. Women with recent IPV experiences reported greater service needs than women with more remote IPV experiences, who in turn reported greater need than women without IPV experiences. We conclude that IPV may be associated with ongoing perceived needs for social and legal services among Latina perinatal patients. PMID- 19297890 TI - Prevalence and risk factors of gender-based violence committed by male college students in Awassa, Ethiopia. AB - We sought to determine the prevalence and risk factors for the perpetration of gender-based violence among 1,378 male undergraduate students in Awassa, Ethiopia. A self-administered questionnaire was used to collect information. Nearly a quarter (24.4%; 95% confidence interval [CI] 22.1-26.7) of students admitted perpetrating acts of gender-based violence during the current academic year. Approximately 15.8% (95% CI 13.7-17.9) of students reported physically abusing, and 16.9% (95% CI 14.8-19.0) reported committing acts of sexual violence against an intimate partner or nonpartner. Alcohol consumption, khat use (Catha edulis, a natural stimulant), combined use of alcohol and khat, and witnessing parental violence were risk factors for committing gender-based violent acts. These findings suggest an obvious need for effective prevention programs targeted toward changing social norms on the use of violence. PMID- 19297891 TI - Diagnosis and endoscopic resection of a bleeding duodenal lipoma. AB - The esophagus, stomach, and duodenal bulb are frequent sites for upper gastrointestinal bleeding. The distal duodenum or other areas of the small intestine are much less common locations for blood loss. Here, we report bleeding in a patient from a 4-cm lipoma in the third portion of the duodenum. The diagnosis of lipoma was strongly suspected from the CT scan and endoscopic features. Endoscopic polypectomy was done after epinephrine needle infiltration of the stalk base. Retrieval of the polyp was accomplished using a Roth Net (US Endoscopy, Mentor, OH). We describe our experience of this endoscopic resection technique being used for this rare cause of upper gastrointestinal bleeding. PMID- 19297892 TI - Blazing saddles. PMID- 19297893 TI - ADHD: attention deficit healthcare disorder. PMID- 19297894 TI - The father of UMC: reflections on the genius of David Schultz Pankratz, MD, PhD. PMID- 19297895 TI - Updates in kidney transplantation--University of Mississippi Medical Center. AB - The kidney transplant program at the University of Mississippi Medical Center has made substantial recent improvements with regard to volume of transplants performed, institutional commitment, and service to the citizens of Mississippi with End-Stage Renal Disease. Patients are being discharged earlier, with satisfactory kidney function; with a low risk for return to hemodialysis both short-term and long-term. Laparoscopic donor nephrectomy is currently being emphasized for people wishing to donate a kidney. Future directions include extra renal abdominal organ transplantation, multiple-center collaborations, and an emphasis on basic science and clinical research. PMID- 19297896 TI - Simple rules, complex behavior. PMID- 19297897 TI - The world is too much with us.... PMID- 19297898 TI - Is there a doctor in the house? (There could be another one; it depends on you!). PMID- 19297899 TI - Rural medical practice. PMID- 19297900 TI - Just a general practitioner. PMID- 19297901 TI - The crucifixion revisited. AB - This article is a follow up of an original article by the author on the crucifixion of Jesus published in the March 1989 issue of the JMSMA. The pathogenesis of the death of Jesus is still widely debated with vastly divergent views as to the specific cause of His death. The author's views have been challenged in the literature. As a result, he has done spirometry studies of volunteers hanging on a cross and has a better understanding of the physiology of the terminal events. That information is presented in this update. PMID- 19297902 TI - Teamwork and pain. PMID- 19297903 TI - Fat chance. PMID- 19297904 TI - Premature death. PMID- 19297905 TI - An open letter to fellow physicians on the good news of the Bible. PMID- 19297906 TI - Perspectives on Mississippi's 21st century Physician Workforce Supply: findings from the 2007 MSMD survey. AB - The capacity to recruit and retain physicians to practice in Mississippi has been a perennial concern of the profession. In the first Mississippi Physician Workforce Study (2003) conducted at the height of the state 'malpractice crisis,' Professor Cossman identified several looming threats to effective Mississippi physician recruitment and retention, including a high percentage of physicians who reported they were considering relocation or retirement in the near future. In this article, Street and Cossman report survey findings from actively practicing physicians (N=848) who responded to the second Mississippi Physician Workforce Study (2007 MSMD). This analysis updates perspectives on the physician workforce supply in the aftermath of malpractice legislative reform and Hurricane Katrina. PMID- 19297907 TI - Isolated muscle mass as an initial presentation of sarcoidosis: a case report and discussion. AB - Sarcoidosis is a multisystemic inflammatory disorder of unknown etiology. It can present in various clinical forms. Involvement of muscles is common but isolated muscle mass, the only initial presenting complaint has never been reported. We report a 55-year-old white female who presented with a muscle mass, with no other clinical features of sarcoidosis. She was later found to have hilar lymphadenopathy and muscle biopsy confirmed the diagnosis of sarcoidosis. During clinical follow up patient remained stable without any medical intervention. PMID- 19297908 TI - Mrs. Feldman goes to Washington (you're not paranoid if they really are out to get you). PMID- 19297909 TI - Do we have what it takes? PMID- 19297910 TI - A call to prevent... PMID- 19297911 TI - Milestone transplant gives Pearl resident second chance at life. PMID- 19297912 TI - Stents for chronic venous insufficiency: why, where, how and when--a review. AB - Venous stenting has introduced a minimally invasive and safe technique that can be performed on an outpatient basis with little downtime for the patient. It is applicable in a wide spectrum of patients with chronic venous disease with disabling symptoms. Long-term patency and clinical outcome are excellent even when associated reflux is present and left untreated. The need for open surgery to correct reflux or obstruction has been drastically reduced; however, such procedures can still be carried out later in the event of stent occlusion or failure to relieve symptoms. PMID- 19297913 TI - An interview with 2008-09 MSMA president J. Patrick Barrett, MD. Interview by Karen A. Evers. PMID- 19297914 TI - In response to: "Has our MSMA been hijacked?". PMID- 19297915 TI - Hypocalcemia following hemithyroidectomy. AB - Hypocalcemia as a manifestation of hypoparathyroidism is a well recognized complication of total thyroidectomy. Less well recognized is the possibility of hypocalcemia following subtotal thyroidectomy. A prospective observational series of 107 patients undergoing hemithyroidectomy was evaluated by preoperative serum calcium and albumin determinations as well as serum calcium determinations at one and seven days postoperatively. There was a significant decrease in mean calcium concentration from the preoperative determination (9.1 mg/dL) to postoperative day one determination (8.4 mg/dL) and then a significant mean increase in the calcium concentration from postoperative day one to day seven (9.0 mg/dL). There was no statistical difference between the mean preoperative calcium concentration and the day seven mean calcium concentration. Additional analyses failed to show a significant relationship between repeated calcium determinations and either disease pathology (benign versus malignant), sex, or race. At one week post surgery, patients' serum calcium normalized. Thus the need for calcium replacement or prolonged hospitalization was generally not necessary. The possible exceptions are patient's with low or low normal calcium preoperatively, unknown status of their parathyroid glands (i.e., previous thyroid surgery), devascularization/resection of parathyroid tissue at the time of surgery and patients undergoing completion thyroidectomy. PMID- 19297916 TI - Mississippi physicians must work to preserve tort reform this election season. PMID- 19297917 TI - When should serum magnesium be measured prior to non-cardiac surgery? AB - While prospective data is scant, the advisability of magnesium measurement appears to be convincing in selected circumstances. Electrolyte abnormalities, mainly hypokalemia and hypomagnesemia, are associated with an increased risk for developing cardiac arrhythmias, however, conflicting opinions remain. If performed on all patients undergoing non-cardiac surgery, magnesium measurement would add considerably to health care costs; however, if limited to patients identified as high risk by the above parameters, the additional cost per patient is in keeping with other standard tests. Given the low cost of testing and replacement, preoperative magnesium measurement appears reasonable in patients with structural heart disease, diabetes, chronic alcoholism, and chronic (loop) diuretic use. When high risk patients are undergoing high risk operations, the low cost of measurement and replacement appears particularly cost-effective, whether it reduces morbidity and prolonged hospitalization from atrial fibrillation or mortality from sustained ventricular arrhythmias. Until additional data becomes available, use of this simple test in patients likely to manifest hypomagnesemia who are at elevated risk of cardiac tachyarrhythmias and cardiac complications following non-cardiac surgery seems prudent. PMID- 19297918 TI - Cryptic abdominal pain. PMID- 19297919 TI - Suicides in Mississippi: recent trends. PMID- 19297920 TI - Physician suicide--a problem for our time. PMID- 19297921 TI - Back to business. PMID- 19297922 TI - A rose by any other name... PMID- 19297923 TI - Mississippi: most comprehensive trauma system in the nation. PMID- 19297924 TI - Non-cognitive variables and residency choice. AB - PURPOSE: The objective for this study was to determine a means of using non cognitive variables to target undergraduate efforts towards likely candidates for specific residency choice. APPROACH AND MATERIALS: Over a three year spread, a volunteer cohort of freshmen medical students was assessed for the non-cognitive variable of personality preference using the Myers-Briggs Type Indicator (MBTI) to determine its accuracy in predicting the choice of specialty training. RESULTS: Of the 98 participants, 13 ultimately chose a residency in Obstetrics and Gynecology (OB/GYN). Four had scores which classified them with the personality preference ESFP (Extroverted, Sensing, Feeling, Perceiving), representing a ratio of 3.5 to 1 when compared to students expressing other personality preferences among the total sample of subjects. CONCLUSION: These preliminary findings indicated that classification of personality preference among freshmen medical students is a statistically reliable means of predicting future residency choices. PMID- 19297925 TI - Hypertrophic cardiomyopathy. PMID- 19297926 TI - Leave no stone unturned. PMID- 19297927 TI - Collaboration. PMID- 19297928 TI - The medical tragedy of the commons. PMID- 19297930 TI - The faith of the moneychangers. PMID- 19297931 TI - Inflammatory bowel disease: thinking outside of the intestines (Part 1). AB - Inflammatory bowel disease (IBD) can present itself in many different ways. Although it mainly affects the gastrointestinal tract, it can manifest itself practically anywhere else within the body. These extraintestinal manifestations (EMs) can occur both during active disease and when the disease is quiescent, and in some cases can precede the diagnosis of IBD. It is imperative for physicians to recognize the EMs of IBD since some of them can cause significant morbidity and mortality if left untreated. This article will review the EMs of IBD which involve the hepatobiliary, mucocutaneous, musculoskeletal and ophthalmologic systems. PMID- 19297932 TI - Inflammatory bowel disease: thinking outside of the intestines (Part 2). AB - Inflammatory bowel disease (IBD) can present itself in many different ways. Although it mainly affects the gastrointestinal tract, it can manifest itself practically anywhere else within the body. These extraintestinal manifestations (EMs) can occur both during active disease and when the disease is quiescent, and in some cases can precede the diagnosis of IBD. It is imperative for physicians to recognize the EMs of IBD since some of them can cause significant morbidity and mortality if left untreated. This article will review the EMs of IBD which involve the hematologic, cardiovascular, pulmonary, neurologic and urinary systems. PMID- 19297933 TI - The rash that caused confusion. PMID- 19297934 TI - Change: President Obama's health plan. PMID- 19297935 TI - Risks. PMID- 19297936 TI - Compliance with and understanding advance directives in trainee doctors. PMID- 19297937 TI - Economic impacts of physicians on Mississippi's county economies. PMID- 19297938 TI - Why the confusion? PMID- 19297939 TI - A start toward a healthier Mississippi. PMID- 19297940 TI - Draw your own conclusions. PMID- 19297941 TI - A doctor's adventures in patient land. PMID- 19297942 TI - Anti-vaccine legislation threatens immunity. PMID- 19297943 TI - New mobile field hospital means immediate care for disaster-stricken Mississippians. PMID- 19297944 TI - Hypertension in Mississippi: we can do better. AB - Hypertension prevalence rates in Mississippi are currently the highest in our country. Hypertension treatment and control rates are less than ideal. Our best hope is in prevention of hypertension through healthier lifestyles with more physical activity, fewer calories, and lower dietary sodium intake. A commitment by physicians to overcome clinical inertia and achieve control in most patients will be necessary for us to make progress. PMID- 19297945 TI - Too much of a good thing. PMID- 19297946 TI - Improving the "business of medicine". PMID- 19297947 TI - Daschle's solutions for America's broken health care system. PMID- 19297948 TI - Medicine: where are you going? PMID- 19297949 TI - The physicians' perspective medical practice in 2008. PMID- 19297950 TI - Where I fit in the food chain. PMID- 19297951 TI - Establishment of a mouse model to assess brain neurotransmitter level and learning performance simultaneously following toxic chemical exposure: using in vivo microdialysis and schedule-controlled operant behavior. AB - Most of the volatile organic compounds used in industrial and household products are reported to be toxic. While the effects of toxic chemicals on pulmonary and other systemic functions have been studied substantially, little is known about their effects on higher brain functions, particularly on the events in learning performance. From the particular perspective of environmental health, it is necessary to examine the toxic effects of environmental chemicals on higher brain functions. The aim of the present study is to establish a mouse model for simultaneous assessment of the effect of toluene (150 mg/kg, i.p.) on extracellular glutamate level in the hippocampus, using in vivo microdialysis and learning performance with a procedure of schedule-controlled operant behavior (SCOB). Different patterns of hippocampal glutamate level were observed in control and toluene-treated mice during learning performance. During learning performance in an operant chamber, control mice showed a saturated and persistent increased glutamate level with good learning performance, while toluene-treated mice showed a decreased glutamate level with poor learning performance. This is the first time to establish a new method to assess the effect of environmental chemical exposure on the brain neurotransmitter level during learning performance in an animal model. PMID- 19297952 TI - Emission rates of air pollutants from portable gas ranges and nitrogen dioxide exposure assessment in restaurants. AB - It is important to characterize the emission of air pollutants and suggest an optimum ventilation rate, because the use of portable gas ranges is widespread in houses and restaurants in Korea. Source emission tests were conducted to characterize the emissions of nitrogen oxide (NOx = NO + NO2), carbon monoxide (CO), carbon dioxide (CO2), formaldehyde (HCHO) and total volatile organic compounds (TVOCs) from portable gas ranges in steady-state using a well-mixed chamber. The ranges of emission rates of air pollutants from portable gas ranges were 0.55-0.94 mg/h for NO, 0.35-1.08 mg/h for NO2, 1.21-1.63 mg/h for NOx 1.39 4.21 mg/h for CO, 2430-2970 mg/h for CO2 and 0-0.12 mg/h for TVOCs. The required mean and maximum ventilation rates to control the air pollutants from portable gas ranges was 2.70 m3/h and 3.13 m3/h on the basis of the NO2 emission rate, respectively. The mean concentrations of food service worker and customer exposures to NO2 by use of portable gas ranges in restaurants were 48.2 +/- 21.5 ppb and 64.7 +/- 31.5 ppb, respectively. PMID- 19297953 TI - [The role of law in protection from violent and abusive patients]. AB - What counter-measures can medical professionals take against violent and abusive patients in Japan? The aim of this article is to consider the role of law for protecting medical professionals from such patients in Japan, comparing it with its role in the UK. Medical professionals need to have legal knowledge for avoiding trouble after using such measures as "verbal defense" or "negotiation". When they use their body or objects for defense, they must use them within the limits of the law. Patients sometimes have violent and abusive behavior because of disease or depression from their condition or the environment. If a medical professional fights back against such a patient, is (s)he legally or morally responsible? This is a very important problem. Professional discretion is concerned with balancing patient healthcare and the safety of medical professionals. PMID- 19297954 TI - [The development of a method to evaluate general ward nursing care practices targeted at families of cancer patients--identification and selection of items for evaluation]. AB - As a first step to develop a method to evaluate general ward nursing care practices targeted at families of cancer patients, the current study aimed to identify and select items for evaluation. Forms of nursing care considered to be necessary for the families of cancer patients were extracted from previous literature and categorized. We identified 5 categories, and 20 items were created to reflect the type of nursing care in these categories. A paper questionnaire evaluation composed of the 20 items was given to 77 general ward nurses. Valid responses were obtained from 67 (87.0% response rate). Factor analysis revealed 6 subscales which included "encourage family to participate in patient care", "commitment and adjustment", "adjust nursing care to meet family member needs", "realize the burden on family members", "attend the death", and "respect family decisions". Items for evaluation were refined based on other responses in the questionnaire, and a total of 26 items regarding nursing care were selected for use in the evaluation. PMID- 19297955 TI - [Seborrheic keratosis on the umbilicus: case report and review of Japanese literature]. AB - A 78-year-old man was seen for a tumor on his umbilicus. Clinically, there was a dark brown nodule with thick scales. The excised tumor showed the acanthotic type of seborrheic keratosis. Six cases of seborrheic keratosis arising on the umbilicus have been reported. All of them were of the histologically hyperkeratotic type. Our case is the first report of an acanthotic type of seborrheic keratosis on the umbilicus. Seborrheic keratosis is one of the common skin tumors, but we hesitated in this diagnosis because of its abnormal location. PMID- 19297956 TI - [A case of acalcuous cholecystitis developed after cardiopulmonary resuscitation]. AB - A 47-year-old man was found at his home in a state of cardiopulmonary arrest. His family performed cardiopulmonary resuscitation on him. He was brought to our hospital by ambulance. On arrival, his pupils were dilated and his heart was in a state of ventricular fibrillation. After returning to spontaneous circulation by the cardiopulmonary resuscitation, the electrocardiogram revealed ST elevation at V2-V5. Cardiac catheterizatin revealed a left anterior descending coronary obstruction. Percutaneous coronary angioplasty was performed. On the 26th day after admission, acalculous cholecystitis was found. It was difficult to perform emergent surgery, because the patient was taking an anticoagulant drug. We performed PTGBA (percutaneous transhepatic gallbladder aspiration) on the same day, and the gallbladder inflammation was improved. We consider that PTGBA is an effective treatment for difficult cases of acalcuous cholecystitis. PMID- 19297957 TI - [A case of intraoperative latex anaphylaxis during general anesthesia for rectal cancer resection]. AB - A 72-year-old woman underwent high anterior resection for rectal cancer. Fifteen minutes after the start of the operation, the pressure on the respiratory tract rose, blood pressure and SpO2 dropped, and skin rash, wheezing and ECG disorder appeared. The anesthesiologist suspected latex anaphylaxis and interrupted the operation. The patient was treated immediately for anaphylactic shock, and recovered. Post-operation, she showed positive reaction to latex specific IgE, suggesting that the anaphylaxis was induced by latex allergy. A re-operation was performed under a latex-free environment. No allergic reaction was seen during or after the re-operation. PMID- 19297958 TI - [Physical therapy for four patients with severe axonal Guillain-Barre syndrome in acute stage]. AB - We describe four male patients suffering from quadriplegia caused by axonal Guillain-Barre syndrome. They were diagnosed with axonal Guillain-Barre syndrome based on their clinical course and electrophysiological findings. Three patients had severe pain, two had marked elevation of serum creatine kinase, one had finger contracture, and two required mechanical ventilation. Two of the patients recovered during the period from nadir to discharge in the Hughes functional grading scale and Barthel index. A physical therapist should understand the characteristics of severe axonal Guillain-Barre syndrome in an acute stage, and consider the psychological status of the patients. PMID- 19297959 TI - [Planning and carrying out of health events: advice for success]. AB - Individual self-motivating efforts and environmental arrangement by business organizations are essential to prevent lifestyle related disease, because factory workers go through many stages of life in the workplace. It is desirable to discuss which health activities that come into operation by occupational and health staff's enthusiasm and inventiveness are best outside the framework of the company. In this article, we discuss advice for success in planning and carrying out health events from perspectives such as previous problems, projects, timing, location, numbers of participants, human resources of the host, estimated cost, selection of channel of advertisement and analyses of data. PMID- 19297960 TI - [Lifestyle concerning physical activity and mental health in university students]. AB - To examine the relationship between lifestyle concerning physical activity and mental health, 372 university students filled out a questionnaire containing lifestyle patterns, subjective physical symptoms and subjective mental symptoms. As a result, a good lifestyle concerning physical activity was related to preferable mental conditions. The joint association of the lifestyle and mental health was more evident in female students than in male students. Consequently, we propose that educational advice for the improvement of lifestyle through physical activity is an important health strategy for university students. PMID- 19297961 TI - Changing paradigms and challenging assumptions: redefining quality and NCLEX-RN pass rates. PMID- 19297962 TI - Comparison of syllabi expectations between faculty and students in a baccalaureate nursing program. AB - This study aimed to explore and compare expectations of syllabi between students and faculty in a university baccalaureate nursing department. Knowing what students expect from syllabi can lead to improved student success and may reduce faculty time in clarifying class policies. Faculty and nursing students from eight semesters volunteered to complete a survey exploring syllabi definitions, pertinent content, and the importance of student involvement in syllabi development. The findings suggest there are differences between faculty and student perceptions regarding important syllabi content. Students wanted syllabi that provided the nuts and bolts of how to accomplish each assignment and course requirement most efficiently. Faculty preferred information about student behavior, such as student conduct, participation, and attendance rules. Adult Learning Theory was used to explain these differences. This article points out that faculty may not be as in touch with the needs of adult learners as they claim to be. PMID- 19297963 TI - Exposing shame and its effect on clinical nursing education. AB - Shame is identified as a universal dynamic in education. Brain-based learning theory suggests negative emotions like shame have a powerfully detrimental effect on learning. Shame theory may explain why students have difficulty identifying with professional nursing culture. Yet shame has neither been directly described nor referred to in the context of clinical nursing education. Accordingly, the aim of this article is to raise awareness among nurse educators about shame and its potential effect on students' ability to learn in clinical nursing education. This article examines shame in its many manifestations; the power to shame inherent in the clinical context; the consequences of shame on students' ability to learn; and, finally, the knowledge, skills, and attitudes needed by nurse educators to heal and prevent shaming in clinical nursing education. PMID- 19297964 TI - Alliance for Clinical Education (ACE): setting community standards through collaboration. AB - Today's clinical nursing education challenges center around managing complex issues related to clinical placements. The increased numbers and varying levels of nursing students, acuity and intensity of the clinical setting, and rise of regulatory and accrediting mandates provided impetus for the establishment of the Alliance for Clinical Education (ACE). This article describes community standards established by partnering educational programs and clinical agencies in the Denver, Colorado, and surrounding areas. ACE is successfully confronting the challenges of effectively using scarce clinical placements, validating student orientation mandates, and facilitating the availability and development of qualified clinical faculty. Examples of tools and resources created by ACE to provide uniformity in meeting mutual expectations and requirements are described. The power of collaboration and sharing expertise is described by current initiatives, which reflect the ability to influence policy and shape community standards to support excellence in clinical nursing education. PMID- 19297965 TI - Alternate item types: continuing the quest for authentic testing. AB - Many test developers suggest that multiple-choice items can be used to evaluate critical thinking if the items are focused on measuring higher order thinking ability. The literature supports the use of alternate item types to assess additional competencies, such as higher level cognitive processing and critical thinking, as well as ways to allow examinees to demonstrate their competencies differently. This research study surveyed nurses after taking a test composed of alternate item types paired with multiple-choice items. The participants were asked to provide opinions regarding the items and the item formats. Demographic information was asked. In addition, information was collected as the participants responded to the items. The results of this study reveal that the participants thought that, in general, the items were more authentic and allowed them to demonstrate their competence better than multiple-choice items did. Further investigation into the optimal blend of alternate items and multiple-choice items is needed. PMID- 19297966 TI - Health assessment in a 2-week intensive format for second-degree baccalaureate nursing students. AB - The proliferation of second-degree baccalaureate nursing programs in the United States has led to a critical need for effective strategies to meet the unique learning needs of this group of students. This article describes an innovative approach to delivery of a health assessment course for accelerated nursing students. PMID- 19297967 TI - Faculty achievement tracking tool. AB - Faculty development and scholarship is an expectation of nurse educators. Accrediting institutions, such as the Commission on Collegiate Nursing Education, the National League for Nursing Accrediting Commission, and the Higher Learning Commission, all have criteria regarding faculty achievement. A faculty achievement tracking tool (FATT) was developed to facilitate documentation of accreditation criteria attainment. Based on criteria from accrediting organizations, the roles that are addressed include scholarship, service, and practice. Definitions and benchmarks for the faculty as an aggregate are included. Undergoing reviews from different accrediting organizations, the FATT has been used once for accreditation of the undergraduate program and once for accreditation of the graduate program. The FATT is easy to use and has become an excellent adjunct for the preparation for accreditation reports. In addition, the FATT may be used for yearly evaluations, advancement, and merit. PMID- 19297968 TI - A collaborative approach to group experiential learning with undergraduate nursing students. AB - Because many nursing responsibilities are accomplished in groups, undergraduate nursing curricula need to prepare students in the skills required to function effectively in groups. This article describes an interdisciplinary, group experiential learning approach used with baccalaureate nursing students as group participants and graduate counseling psychology students as facilitators. This teaching method provides learning to enhance the knowledge and skills of students preparing for both professions. PMID- 19297969 TI - The importance of nursing research. AB - Nursing research has a tremendous influence on current and future professional nursing practice, thus rendering it an essential component of the educational process. This article chronicles the learning experiences of two undergraduate nursing students who were provided with the opportunity to become team members in a study funded by the National Institute of Nursing Research. The application process, the various learning opportunities and responsibilities performed by the students, and the benefits and outcomes of the experience are described. The authors hope that by sharing their learning experiences, more students will be given similar opportunities using the strategies presented in this article. Nursing research is critical to the nursing profession and is necessary for continuing advancements that promote optimal nursing care. PMID- 19297970 TI - Faculty retirement: stemming the tide. AB - The shortage of nursing faculty is becoming a crisis in schools of nursing and is contributing to the shortage of qualified graduates to add to the nursing workforce. Aging and retirement of existing faculty contribute to the crisis. Creating retention initiatives may encourage faculty to postpone retirement. This article explored the conditions and situations which would entice faculty to continue working beyond their intended retirement date. Faculty identified workload and responsibilities, available resources, personal and professional characteristics, and work environment as areas that could be developed into incentives to remain in the faculty workforce. PMID- 19297971 TI - Teaching prescriptive authority through active learning. PMID- 19297972 TI - Pesticide usage pattern in tea ecosystem, their retrospects and alternative measures. AB - Tea is a perennial plantation crop grown under monoculture providing favorable conditions for a variety of pests. The concept of pest control has undergone a considerable change over the past few decades. In recent years there has been a greater dependence on the use of pesticides (7.35-16.75 kgha(-1)) with little importance laid on other safe control methods for the management of tea pests. Due to this practice, the tea pests showed a higher tolerance/ resistance status due to formation of greater amount of esterases, glutathione S-transferase and acetylcholinesterase. Thus, over reliance on pesticides end up with pesticide residue in made tea (DDT - 10.4-47.1%; endosulfan - 41.1-98.0%; dicofol- 0.0 82.4%; ethion - 0.0-36.2%; cypermethrin - 6.0- 45.1%). The growing concern about the pesticide residue in made tea, its toxicity hazards to consumers, the spiraling cost of pesticides and their application have necessitated a suitable planning which will ensure a safe, economic as well as effective pest management in tea. At present it is a global concern to minimize chemical residue in tea and European union and German law imposed stringent measures for the application of chemicals in tea and fixed MRL values at < or = 0. 1 mgkg(-1) for the most commonly used pesticides which will not be met out in the real practice and has been a major constraint to tea exporting countries like India. In order to regulate the situation of the Indian market at global level, central insecticide board and prevention of food adulteration regulation committee have reviewed the MRL position for tea and has recommended 10 insecticides, 5 acaricides, 9 herbicides and 5 fungicides for use in tea and issued the tea distribution and export control order 2005 which will help the country to limit the presence of undesirable substances in tea. This review attempts to provide the readers with a comprehensive account of pesticide use in North East in tea, surveillance report of the European community regarding the residue level in Assam and Darjeeling tea, recent amendments by international and national regulatory bodies, revised MRL values of pesticides in tea, an update about the current strategies for the management of tea pests with more focus on the use of biological control agents and a possible beneficial role or judicious use of chemical pesticides in complement with other alternative measures to achieve optimum effects in terms of limiting agricultural input, lowering production costs, reducing environmental contamination and the effect on non-target organisms, delaying the development of resistant pest biotypes and above all minimizing the pesticide residues in tea to increase the exports. PMID- 19297973 TI - Estimation of Cochlodinium polykrikoides growth potential using a dialysis membrane. AB - We developed a test to measure the growth potential of C. polykrikoides using a dialysis membrane and artificial seawater. Nitrite nitrogen and inorganic phosphorus in the medium were almost completely removed when the medium was dialyzed against artificial seawater for five or more 6-hour cycles using a dialysis membrane (Spectrum's Spectra/Por 7 Membrane) with a molecular-weight cut off of 50,000, regardless of the presence of C. polykrikoides. The phytoplankton grew well even after dialysis. To estimate the growth potential of C. polykrikoides, a minimum initial concentration of > 100 cells/ml is required. Methods using short-term starvation culturing of C. polykrikoides to measure growth potential were determined to be ineffective; instead, controlled tests using artificial seawater are recommended. The dialysis membrane used in this study can also be employed to measure the algal growth potential of other phytoplankton species. PMID- 19297974 TI - Decolorisation of synthetic dyes and textile wastewater using Polyporus rubidus. AB - Effluent from textile industries were treated with enzyme from white rot fungi isolated from outskirts of Mumbai and identified as Polyporus rubidus in our laboratory. Decolorisation of 4 Reactive dyes commonly found in the effluents such as Reactive bue, Reactive orange, Ramazol black and Congo red was examined by treatment with enzyme from Polyporus rubidus. Treatment of effluent was done in a laboratory scale bioreactor constructed with laccase immobilized Na-alginate beads. Greater than 80% of dyes were degraded within 5 days under stationary incubation conditions. The enzyme had a maxmimum activity of 17.1U after 3 days and was found to be secreted extracellularly by Polyporus rubidus. In this study the Polyporus rubidus has been reported for the first time to have laccase activity offering a promising possibility to develop an easy and cost effective method for degradation of dangerous dyes. PMID- 19297975 TI - Acute toxicity of organophosphate insecticide, dichlorvos in relation to selected water hardness for the freshwater zooplankters. AB - Toxicity of organophosphate insecticide, i.e. dichlorvos has been investigated in relation to selected water hardness for the freshwater zooplankters such as Moina, Daphnia, cyclops and nauplii. Results revealed that dichlorvos was highly toxic to the tested zooplankters, as the LC50 values are noticed in ppb. Trend of sensitivity for different zooplankters to dichlorvos was recorded as Moina < Daphnia < nauplii < cyclops. The LC50 were found to change significantly with the change in water hardness. The zooplankters were more susceptible to dichlorvos at water hardness of 275 mgl(-1) as compared to water hardness of 540 mgl(-1) as revealed from low LC50 values. The range of safe dischargeable concentrations (1.063-1.137ppb) were too low as compared to harmless or safe concentrations (43.895-89.194 ppb) for the zooplankters at both the hardness. PMID- 19297976 TI - Calli cultures from Abies equi-trojani (Aschers et Sinten) and changes in antioxidant defense system enzymes. AB - This study was conducted to explain difficulties of indirect regeneration of forest trees in tissue culture conditions. For this purpose, changes of antioxidant defense system enzymes; superoxide dismutase (SOD), peroxidase (POD) activities were determined during calli formation on young apical shoots of Abies equi-trojani (Aschers et Sinten). Young apical shoots were collected from naturally growing trees and cultured on two different media; Murashige and Skoog (MS) and McCown Woody plant medium (WPM) supplemented with growth regulators benzyl amino purine (BAP), 2,4-dichloro phenoxy acetic acid (2.4-D), kinetin (Kn) in various concentrations for callus induction. WPM media containing 1 mg ml(-1) BAP and 1 mg ml(-1) 2,4-D gave the best calli induction ratio (74%) between tested combinations. POD and SOD enzyme activities were measured both on young shoot explants and 10 day-old calli derived from these explants. POD and SOD enzyme activities were higher being 81.02% and 74.82%, respectively on calli when compared to shoots. The results showed that culture stress tolerated with increased antioxidant enzyme activities could be considered as protective physiological responses in calli cells. PMID- 19297977 TI - Possible modulating action of plant infusion of Ocimum sanctum L. on chromosomal aberrations and sister chromatid exchanges induced by chlormadinone acetate in human lymphocytes in vitro. AB - Chlormadinone acetate (CMA) is a synthetic progesterone analogue. It has its usage in oral contraceptives formulations and also for estrous synchronization of animals. The aim of the present study is to study the anti- genotoxic activity of the plant infusion against the CMA induced genotoxic damage on cultured human lymphocytes, using chromosomal aberrations and sister chromatid exchanges (SCFs) as parameters. For chromosomal aberration analysis, the treatment of 40 microM of CMA was associated with 4.33% abnormal metaphases. The treatment of 40 microM of CMA, separately with 1.075 x 10(-4), 2.125 x 10(-4) and 3.15 x 10(-4) gm l(-1) of plant infusion results in the reduction of the number of abnormal metaphases i.e. 2.67%, 2.00% and 1.67% respectively. For sister chromatid exchange analysis, the frequency of sister chromatid exchange per cell (SCE(S)/Cell) for the treatment of 40 microM of CMA was 6.43. The treatment of 40 microM of CMA, separately with 1.075 x 10(-4), 2.125 x 10(-4) and 3.15 x 10(-4) gm l(-1) of plant infusion results in the significant reduction of the frequency of SCE(S)/Cell i.e. 3.76, 3.01 and 2.94, respectively, as compared to the CMA (40 microM) treatment alone (6.43). The used dosages of plant infusion did not increase chromosomal aberrations and sister chromatid exchanges at significant level as compared to the untreated. The results of the present study suggest that the plant infusion per se does not have genotoxic potential, but can modulate the genotoxicity of chlormadinone acetate in human lymphocytes in vitro. PMID- 19297978 TI - The effect of nitrogen and potassium fertilizers on growth parameters and carbohydrate contents of sweet sorghum cultivars. AB - Sweet sorghum is tolerant to high temperature and drought and can be considered as an alternative crop to sugar beet and maize in Iran. In this study, the effects of nitrogen and potassium fertilizers on growth parameters including stem height, stem diameter, stem fresh weight, total fresh weight; carbohydrate contents including total sugar, brix value, sucrose content and purify; and juice extract of two sweet sorghum cultivars were determined. Three rates of N fertilizer (0, 90, 180 kg urea ha(-1)) and two rates of K fertilizer (0 and 50 kg potassium sulfate ha(-1)) assigned as main plots and two sweet sorghum cultivars (Rio and Keller) as subplots. Growth parameters at soft dough and physiological maturity stages and carbohydrate contents at physiological maturity stage were determined. Results showed that application of 180 kg urea ha(-1) as compared to control at physiological maturity significantly (p < 0.01) increased stem height (12.65%), stem fresh weight (24.57%), total fresh weight (78.22%), total sugar (39.25%), sucrose content (9%) and juice extract (34.96%). Application of 50 kg potassium sulfate ha(-1) increased (p < 0.05) stem fresh weight (24.33%), total fresh weight (25.44%), total sugar (10.50%), and juice extract (9%) at physiological maturity. The highest growth parameters, carbohydrate contents and juice extract were obtained with the application of 180 kg urea ha(-1) and 50 kg potassium sulfate ha(-1) using cultivar (cv) Keller. The best results were taken with the application of both fertilizers. PMID- 19297979 TI - Toxicity of cadmium on the growth and yield of Solanum melongena L. AB - Heavy metal cadmium is biomagnified through food chain and causes Itai-Itai disease in human. The present investigation reports the results of the effect of cadmium on seed germination, germination relative index (G.R.I.), seedling growth, chlorophyll stability index (CSI) and yield of Solanum melongena L. cv Pusa uttam. Effect of different concentrations of heavy metal cadmium (CdCl2) in Hoagland's nutrient solution (10(-2) M, 10(-4) M, 10(-5) M and 10(-8) M) were employed for all seedling and physiological parameters of brinjal. Cadmium showed toxic effects at high concentrations 10(-2) M but promotory at lower concentration (10(-8) M) with regard to growth and yield. PMID- 19297980 TI - Growth and periodicity of cyanobacterial bloom in a polluted pond of Agra city. AB - Periodical changes in physico-chemical properties and cyanophycean bloom in a pond ofAgra city during the year 2005 and 2006 were studied. It was recorded that the pond water was polluted throughout the study period and this was closely associated with Cyanobacterial bloom. During summer (March to June), the pond water was highly polluted, as it was rich in various nutrients and phosphates in particular. The pH of pond water was high with low DO and this was associated with the highest peak of water bloom. On the other hand, during winter (November to January), the bloom was at its lowest. The quantity of various nutrients in the pond water was low and the pH was also low but DO was high. Bloom consisted of eighteen members of Cyanophyceae of which Oscillatoria limosa, O. formosa, O. splendida, Anabeana iyengarii, Microcystis arruginosa and Nostoc muscorum were observed throughout the year. On the other hand, Oscillatoria agardhi, and O. tenuis were found only in winters, while O. annae in rainy season only. Similarly, Oscillatoria brevis, Anabaena flosaquae, Aphanizomenon sp. and Lyngybea birgei were found in abundance in summer. PMID- 19297981 TI - The immune response in catfish, Mystus gulio. AB - Light microscopic studies of the immune response of Mystus gulio were carried out. Antigen binding cells have been detected by plaque-forming cell (PFC) assay test in the spleen, head-kidney (HK) and thymus. Among these three organs, the HK is more pronounced in its response, and it is compared, on the basis of its histology of higher vertebrates. The peak response after primary and secondary immunizations was on day 7, in both circulating blood and the immune organs. The results suggest that HK in these fish might be the major organ for antibody secreting cells. The HA (haemoagglutinin) response was also for longer duration but only slightly more intense. The PFC response after the secondary immunization was for much longer duration and much more intense than after the primary immunization. PMID- 19297982 TI - Enhanced crude oil biodegradation and rhamnolipid production by Pseudomonas stutzeri strain G11 in the presence of Tween-80 and Triton X-100. AB - In this study, the growth of sixty-one bacterial strains in crude oil were determined spectrophotometrically at 620 nm. Pseudomonas aeruginosa G1, Pseudomonas fluorescens G6, Pseudomonas stutzeri G11 and Pseudomonas putida G15 were chosen for the study based on the efficiency of crude oil utilisation. At 1% (v/v) crude oil concentration, P. stutzeri G11 strain degraded a maximum of 69%. The percentage of degradation by the P. stutzeri G11 strain decreased from 69% to 59% as the concentration of crude oil was increased from 1% (v/v) to 2.5% (v/v). Strain G11 was selected to determine the effects of surfactants (Tween-80 and TritonX-100) on the biodegradation of crude oil. While strain G11 showed 76% degradation at mineral salts medium (MSM) containing 1% (v/v) crude oil + 1% (v/v) TritonX-100, it showed 61% degradation at MSM containing 2.5% (v/v) crude oil + 2.5% (v/v) TritonX-100. Also, degradation rate of this strain was 96% in the presence of 1% (v/v) crude oil + 1% (v/v) Tween-80, while degradation rate was 48% in the presence of 25% (v/v) crude oil+ 2.5% (v/v) Tween-80. Additionally, we investigated the rhamnolipid production of P. stutzeri G11 strain both in crude oil and in crude oil + two different surfactants (TritonX 100 and Tween-80, separately). These results suggest that surfactants have improved both crude oil degradation and rhamnolipid production and the degradation rates have depended very much on the chemical structure of surfactants. PMID- 19297983 TI - Fluoride contamination status of groundwater in Phulera tehsil of Jaipur district, Rajasthan. AB - Over the last few decades the water quality is deteriorating in arid and semi arid regions. Phulera tehsil is facing the problem of groundwater pollution. In the present investigation, determination of fluoride (F) in drinking water was conducted in (200 samples of) 40 villages of Phulera fehsil having fluoride content more than permissible limits (> 1.5 mg l(-1)). After the pilot survey symptoms of skeletal and gut fluorosis have been found in almost every inhabitant. The water samples were alkaline with pH ranging from 7.05 to 10.16. Electrical conductivity (EC) ranged from 157 micromhoScm(-1) to 1018 micromhoS cm(-1). Calcium hardness (Ca-H) ranged from 10 to 127 mg l(-1). Total hardness (TH) varied from 69 to 572 mg l(-1). Chloride varied from 92.00 mg l(-1) to 1422.00 mg l(-1) and fluoride from 1.20 to 18 mg l(-1). The alkalinity of all water samples were found to be more than the permissible limit. The results envisaged that the quality of ground water of Phulera is very poor, and is not suitable for drinking purpose and can only be used after proper treatment. PMID- 19297984 TI - Effect of shearing and environmental conditions on physiological mechanisms in ewes. AB - The aim of this study was to establish the influence of shearing in dairy sheep during summer season. Several physiological and haematological parameters, which are closely related to thermoregulatory potential, have been investigated with the purpose to assess shearing influence on thermoregulation. Forty dairy sheep, clinically healthy and well-fed, were used. They were divided into two groups of 20 subjects each. Twenty sheep were let unshorn as a control group (Group A), and twenty sheep were shorn (Group B). On each subject of group A and Group B, rectal temperature, respiration and heart rates were recorded and blood samples were collected in order to asses the following parameters: red blood cells, white blood cells, platelets, haematocrit and haemoglobin. All measurements taken on day 0 (before shearing), were repeated after 1, 15, 30, 45 and 60 days after shearing. The statistical analysis, the ANOVA followed by the Bonferroni's test, showed statistical differences between two groups for rectal temperature (F(1.90) = 5.57, p < 0.0001), respiratory rate (F(1.90) = 18.40, p < 0. 0001) and white blood cells (F(1.190) = 7.61, p < 0.0001). These results suggest that the shearing induce adaptative responses in the organism. PMID- 19297985 TI - Ground water quality in environmentally degraded localities of Panipat city, India. AB - Asystematic physico-chemical analysis of the groundwater at 41 different locations in Panipat city (Haryana), India has been taken up to evaluate its suitabilityfor domestic purposes. The data revealed considerable variations in the water samples with respect to chemical composition. For the analyzed water samples pH, EC, TDS, TA, TH, Na+, K+, Ca2+, HCO3-, Cl-, SO4(2-) and F(-) varied from 6.6-7.5, 0.09-3.28 mmhoS cm(-1), 700-2100 mg l(-1), 245-1054 mg l(-1) (as CaCO3), 153-520 mg l(-1) (as CaCO3), 57-560 mg l(-1), 5-22 mg l(-1), 36-95 mg l( 1), 298-1285 mg l(-1), 60-311 mgl(-1), 17-786 mg l(-1), 0.24-9.27 mg l(-1) respectively. All samples have high concentration of dissolved salts and all the samples were hard to very hard. Correlation coefficient "r" analysis has been worked out among different water quality parameters. The study shows a positive and significant, correlation of electrical conductivity with total dissolved salts (r = 0.979), total hardness (r = 0.507), sulphate (r = 0.453), total alkalinity (r = 0.725). Total hardness is positively and significantly correlated with magnesium (r = 0.833) and sulphate (r = 0.687). Where as total alkalinity was found to be positively and significantly correlated with bicarbonate (r = 0.992). Fluoride was higher than permissible limits in most of the samples. PMID- 19297986 TI - Neuroendocrine regulation and pesticidal impact on freshwater crab, Barytelphusa guerini (H. Milne Edwards). AB - The extensive use of pesticides to control agricultural pests poses a serious threat to many non-target organisms of the aquatic environment such as the freshwater crab, B. guerini. The deleterious influence of the pesticide causes physiological, biochemical, histological and such other disorders in the animal exposed. In the present study impact of an organophosphate pesticide, monocrotophos and neuroendocrine regulation on the biochemical contents of hepatopancreas of B. guerini has been studied. Experimental studies revealed that glycogen and protein content decreased in normal crabs when exposed to sub lethal concentrations of monocrotophos, while lipid content was increased. In the ablated and pesticide exposed crabs glycogen, protein and lipid contents decreased. In case of ablated and exposed crabs when injected with eyestalk extract, glycogen and protein contents declined, whereas lipid content hiked. It was observed that glycogen, protein and lipid contents in eyestalk extract injected crabs were similar to those of normal exposed crabs. This indicates the vital role of eyestalk in the regulation of biochemical contents. Histological studies of the hepatopancreas indicate structural changes such as large number of vacuolated cells and phagocytes when exposed to the pesticide. PMID- 19297987 TI - Estimation of amino acids, urea and uric acid in tasar silkworm, Antheraea mylitta Drury. AB - The tasar silkworm, Antheraea mylitta Drury, Andhra local ecorace is an exclusive race of Andhra Pradesh. It is on the verge of extinction due to difficulty of acclimatisation at breeding and rearing stages. As an attempt to protect this race, a method of total indoor rearing has been done. In this context, the estimation of free amino acids, excretory products- urea and uric acid were compared during the fourth and fifth instars of tasar silkworm, reared under outdoor and indoor conditions. The study has revealed that amino acids decreased in the fat body in outdoor and indoor reared larvae in contrast to that in the haemolymph where it has gradually increased from first to third crops. This is an important finding as it reveals that indoor worms seem to adopt proteolytic activity in the haemolymph. Secondly, in the fifth instar the excretory products are more compared to fourth instar in the indoor reared worms. During fifth instar, formation of nitrogenous products lessens as silk synthesis enhances. The present study reveals that decrease in uric acid in fifth instar implies increase in growth rate and silk synthesis in both outdoor and indoor worms. The findings of the present investigation is helpful in the conservation and protection of the A. mylitta, Andhra local ecorace. PMID- 19297988 TI - Effects of zinc on morphology of erythrocytes and spleen in Carassius gibelio. AB - The influence of increased zinc concentrations (0.1, 0.5, 1.0, 1.5 and 2.0 mg(-1) ZnSO4 x 7H2O) on the total number and the morphology of the erythrocytes, as well as the processes related to their formation and destruction in the spleen of Carassius gibelio were investigated ex situ. It was found that zinc concentrations caused pathological alterations in the erythrocytes that were not identical in the different concentrations-poikilocytosis; ruptures in cell membranes in the concentrations of 0.5mg(-1) and 1.5 mg(-1); cells with double nuclei (symplasts); in the concentration of 1.0 mg(-1); in the highest concentrations (1.5 mg(-1) and 2.0 mg(-1)) presence of erythrocytes at initial stage of atypical mitotic division. Against the background of those various alterations, the total number of the erythrocytes in the peripheral blood increased simultaneously with the increase of zinc concentrations (p < 0.001). Morphological alterations in the spleen were also observed, indicating a compensational tendency against the toxic influence of zinc upon the fish erythrocytes-hyperplasia of the red pulp and lack of hemosiderin. These results show that the alterations in the total number and the morphology of the erythrocytes are connected with the relevant compensatory histopathological alterations in the spleen. The use of the ascertained alteration could be valuable in monitoring zinc-polluted waters. PMID- 19297989 TI - EDTA enhances lead uptake and facilitates phytoremediation by vetiver grass. AB - Vetiver grass (Vetiveria zizanioides) has strong and dense root system and is a potential phytoremediator plant since it can tolerate a wide range of climatic conditions and grow well in soils contaminated with heavy metals. Soil was artificially contaminated by lead (20 mgl(-1)) during field trials. Four concentration of EDTA (Ethylene diamine tetra acetic acid-disodium salt) solution i.e. 0, 3, 5 and 10 mmol kg(-1) were added to soil prior to harvesting, to study the influence of EDTA solution on phytostabilization by vetiver grass. Results showed that the concentration of lead in roots of vetiver is significantly increased after EDTA solution (5 mmol kg(-1)) application. However, high concentration of EDTA (10 mmol kg(-1)) does not show such significant increase. The toxicity of highly contaminating metal did not affect the growth of vetiver grass significantly but a slight decrease in parameters studied was noticed. No stress symptoms were observed in vetiver plants. Results of present study reveal that vetiver could be considered as a potential phytoremediator for lead contamninated site. PMID- 19297990 TI - Isolation and characterization of high quality DNA from marine benthic macroalgae. AB - The isolation of high quality DNA is essential for many molecular biology applications including polymerase chain reaction (PCR) and endonuclease restriction digestion based techniques. An easy and inexpensive protocol has been developed for extracting genomic DNA from seven species of algae viz. Lola capillaries, Enteromorpha intestinalis, Ulva lactuca and Rhizoclonium sp belonging to Chlorophyceae, Catenella nipae, Polysiphonia mollis belonging to Rhodophyceae and Dictyota ceylanica belonging to Phaeophyceae group were collected from the coastal regions of Sunderban delta in West Bengal, India dominantly growing on mud flats, bark of different mangrove trees, pneumatophores, stilt roots, concrete surfaces, wooden and bamboo poles, sides of the boats and other water vehicles inundated during high tides. The DNA was found suitable for restriction endonuclease digestion and PCR amplification with randomely amplified polymorphic DNA (RAPD) primers. The A260/A280 ratio of 1.15 0.14 to 1.94 indicated little contamination from proteins and polysaccharides. The PCR amplification with RAPD primers showed its suitability in PCR based techniques and the restriction digestion with Eco RV confirmed its suitability for hybridization based techniques. The protocol is equally good for isolating DNA from both fresh as well as preserved materials. PMID- 19297991 TI - Effects of plant lectin from cobra lily, Arisaema curvatum Kunth on development of melon fruit fly, Bactrocera cucurbitae (Coq.). AB - The lectin from tubers of cobra lily, Arisaema curvatum Kunth was purified by affinity chromatography using asialofetuin-linked amino activated porous silica beads. The concentration dependent effect of lectin was studied on second instar larvae (64-72 hr) of Bactrocera cucurbitae (Coq.). The treatment not only resulted in a significant reduction in the percentage pupation and emergence of the adults from treated larvae but it also prolonged the remaining larval development period. A very low LC50 value, 39 mgl(-1) of lectin was obtained on the basis of adult emergence using probit analysis. The activity of three hydrolase enzymes (esterases, acid and alkaline phosphatases), one oxidoreductase (catalase) and one group transfer enzyme (GSTs: Glutathione S-transferases) was assayed in second instar larvae under the influence of the LC50 of lectin at increasing exposure intervals (0, 24, 48 and 72 hr). The Arisaema curvatum lectin significantly decreased the activity of all the enzymes except for esterases, where the activity increased as compared to control at all exposure intervals. The decrease in pupation and emergence as well as significant suppression in the activities of two hydrolases, one oxidoreductase and one GST enzyme in treated larvae of B. cucurbitae indicated that this lectin has anti-metabolic effect on the melon fruit fly larvae. PMID- 19297992 TI - Effect of sublethal exposure of Cartap on hypothalamo-neurosecretory system of the freshwater spotted murrel, Channa punctatus (Bloch). AB - In order to record the effect of carbamate pesticide on hypothalamus of Channa punctatus, fish were exposed to sublethal concentration (0.18 mg l(-1), 30% LC50 for 96 hr) of Cartap for 24, 48, 72 and 96 hr under static bioassay condition. Hypothalamo-neurosecretory complex of the murrel consisted mainly of nucleus preopticus (NPO), nucleus lateralis tuberis (NLT) and their axonal tracts. NPO is a paired structure situtated on either side of the third ventricle anterodorsal to the optic chiasma and looked inverted L-shape in the sagittal section. NPO is morphologically divisible into a dorsal pars magnocellularis (PMC) consisting of large neurons and ventral pars parvocellularis (PPC) formed of smaller neurosecretory cells. NLT cells are distributed in the infundibular floor adjacent to the pituitary stalk. Sublethal Cartap treatment induced an initial hypertrophy of the neurosecretory cells of NPO and NLT followed by loss of staining affinity as well as varying degrees of cytoplasmic vacuolization and necrosis. Herring bodies (HB) were also encountered in the neurohypophysis of the treated fishes. PMID- 19297993 TI - Household participation in recycling programs: a case study from Turkey. AB - This study investigates the underlining factors that motivate households to participate in a pilot source separation and recycling program in Turkey. The data of this research were collected from randomly selected households in the program area via face to face interviews based on an inclusive questionnaire. The results of logistic regression analysis show that having sufficient knowledge regarding recycling and the recycling program is the most statistically significant factor in determining whether a household will participate in recycling. The results also imply that some of the socio-economic and demographic characteristics of household hypothesized to affect the household decision to participate in recycling, in the research framework, are not significant. PMID- 19297994 TI - Impact of differential feeding on the growth and development of Helicoverpa armigera (Hubner). AB - The growth and development of Helicoverpa armigera was observed in the laboratory by feeding them on different foods viz; leaf, flower and fruit of lady finger, cotton, pigeon pea and chick pea. Based on the food ingested, food digested and food excreted as well as on weight and size of the larvae and the duration of larval period, the fruit of chick pea was found to be the most suitable food for H. armigera development, as the food ingesta and food digesta of the larvae on pigeon pea were more than on the other plants. On the pigeon pea pod the larval growth and development was fast and larval duration was short. Next to the fruit, the larvae preferred the leaf of lady finger and cotton and flower of pigeon pea. The results suggested that the larval growth and development was dependent on the feed i.e. both on the part and the type of the plant. PMID- 19297995 TI - Industrial application of keratinase and soluble proteins from feather keratins. AB - Feather keratin is highly resistant to degradation, but some keratinase producing microorganisms can easily degrade these insoluble keratins. These keratinase producing species have an important application in removal of poultry waste and recycled into valuable byproduct. Bacillus sp was screened from soil samples of slaughterhouse and poultry farm area using azokeratin medium. Highest keratinase activity (122.5 KU ml(-1)) was observed at 8.0 pH. Submerged fermentation was carried out at 8.0 pH up to 5th days. On 4th day enzyme production was highest (140 KU ml(-1)) with 1% feather (w/v). Crude protein content was high on day 5, around 1.44 mg ml(-1). 75% of filtrate was found to be crude protein. The molecular weight of this keratinase was 32 kDa by SDS-PAGE. The crude protein from feather has of high nutrient value and could be used as animal feed for livestock and fish feed in aquaculture. PMID- 19297996 TI - Effect of sewage water on seed germination and vigour index of different varieties of groundnut (Arachis hypogaea L.). AB - The study has been focused on the investigation on ground nut (Arachis hypogea) fields influenced by sewage water. Sewage water sampled in and around Mysore city and analyzed forphysicochemical parameters. Different concentrations such as 1, 5, 10, 25, 50, 70 and 100% of sewage water on seed germination and vigour index of Arachis hypogea verities such as DH - 2 - 30, ICJS - 11, JL - 24, K - 134, TMV - 2 and VRI - 2 were studied. From the recorded observation it is concluded that the sewage water diluted to 25% concentration for irrigation of groundnut enhances germination percentage and vigour index in K- 134 variety which is more susceptible than other tested varieties. PMID- 19297997 TI - Larval susceptibility of Aloe barbadensis and Cannabis sativa against Culex quinquefasciatus, the filariasis vector. AB - Larvicidal potential of petroleum ether, carbon tetrachloride and methanol extracts of Aloe barbadensis and Cannabis sativa has been investigated against Culex quinquefasciatus. Among the extracts examined, Carbon tetrachloride extract (Cte) of Aloe barbadensis was the most effective with LC50 values of 15.31 and 11.01 ppm after 24 and 48 hr of exposure, respectively followed by pertoleum ether extract (Pee) of A barbadensis, Cte of C. sativa, methanol extract (Mee) of A. barbadensis, methanol and petroleum ether of C. saliva, LC, being 25.97, 88.51, 144.44, 160.78 and 294.42 ppm affer 24hr and 16.60, 68.69, 108.38, 71.71 and 73.32 ppm after 48 hr of post treatment, respectively. Cte of both the plants exhibits potential larvicidal activity and can be used as ecofriendly alternative in the management of the filariasis vector, Culex quinquefasciatus. PMID- 19297998 TI - Seasonal variations in physico-chemical characteristics of Pichavaram mangroves, southeast coast of India. AB - All the physico-chemical parameters such as temperature, salinity pH, dissolved oxygen and nutrients like nitrate, nitrite, inorganic phosphate and reactive silicate were studied in Pichavaram mangroves, southeast coast of India, for a period of two years (April 2000-March 2002). Air and surface water temperatures varied from 27 degrees C to 38 degrees C and from 26 degrees C to 37 degrees C. Salinity varied from 3.0 per thousand to 33.0 per thousand and the pH ranged between 7.2 and 8.2. Variation in dissolved oxygen content was from 2.4 to 5.0 ml l(-1). Concentrations of nutrients viz nitrates (9.50 to 32.12 microM), nitrites (1.07 to 5.99 microM), phosphates (0.73 to 2.36 microM) and reactive silicates (22.1 to 89.8 microM) also varied independently. PMID- 19297999 TI - New synthetic precocenoids as potential insect control agents. AB - Ageratochromes or precocenes are known for their insect growth regulating (IGR) activity. The present investigation was taken up with an objective to look for the lead structure in these compounds which can be elaborated synthetically to obtain useful growth regulators for practical purposes. With this in mind, some variants of precocenes were synthesized in the laboratory and tested for their toxicity and growth regulating activity using red cotton bug Dysdercus koenigii as the test insect. Most of the precocenoids showed toxicity of various degree and metamorphic derangements to different extents. Adults emerging from treated nymphs could not complete the normal life span. Among the compounds tested 8 acetyl-7-hydroxy-5-methoxy-dimethylchromene (alloevodinol) was more toxic and also showed developmental defects at very low dose such as 0.5 mg l(-1)/nymph. Precocene II (6, 7-dimethoxy-2, 2-dimethylchromene) was used as the standard compound. It was the least toxic and showed effects at 30 mg l(-1)/nymph. PMID- 19298000 TI - Effect of salinity and temperature on the germination of Spergularia marina seeds and ameliorating effect of ascorbic and salicylic acids. AB - Spergularia marina (Caryophyllaceae) is a halophytic species and widely distributed among the sea shores of Turkey. Its seeds maybe unwinged or winged. Laboratory experiments were conducted to determine the effects of salinity (0, 50, 100 and 500 mM NaCl) and temperature (10, 15, 25, 30, 35 degrees C) on seed germination. S. marina showed 73.3% germination in non-saline controls at 25 degrees C. No germination occurred at 30 degrees C and 35 degrees C. Seed germination of Spergularia marina was significantly affected by salinity levels, temperature and their interactions. Maximum final germination percentage occured under the lowest salinity condition (50 mM) and distilled water at 25 degrees C. Seed germination was completely inhibited by 500 mM NaCl, although seed germination rate was not affected by salinity. Recovery germination was greatest in 500 mM and at 25 degrees C. The rate of germination was significantly affected by temperature (p < 0.01). The germination percentage of the seeds pretreated with 40 mM and 60 mM L-ASA in 50 mM and 100 mM NaCl was improved compared with that of untreated L-ASA. Addition of 0.5 mM salicylic acid (SA) improved the germination in 50 mM NaCl. PMID- 19298001 TI - Assessment of teratogenecity and embryotoxicity of sludge from textile industries at Pali (India) in Swiss albino mice exposed during organogenetic period. AB - The present investigation was carried out to assess the teratological effects of in-utero exposure of sludge leachate from textile and dyeing industries located in Pali, Rajasthan. Sludge was collected at the combined effluent treatment plant (CETP). Two groups of 10 pregnant Swiss albino mice each, were given sludge leachate of 1/10 and 1/100 dilutions with water ad libitum from 6th day to 15th day of gestation covering the critical period of organogenesis. Cesarean sections were performed on day 18 of gestation and all foetuses were examined for reproductive and teratological tests. Sludge induced maternal toxicity was evidenced by significant increase in leachate consumption, reduction in body weight gain and reduction in fur of the body. Developmental toxicity was evidenced by a significant decrease in foetal weight per litter increase in the number of resorptions and an increase in total number of foetuses showing bone retardation and skeletal variations (specially of skull, sternebrae and vertebrae). The leachate of the sludge that is being dumped in the open areas of the town Pall seems to elicit teratogenic as well as embryotoxic potential as indicated by the findings of the present investigation. PMID- 19298002 TI - Genetic polymorphism in chemokine CCL22 and susceptibility to Helicobacter pylori infection-related gastric carcinoma. AB - BACKGROUND: Gastric carcinoma is widely considered to be related to Helicobacter pylori infection, and the chemokine (C-C motif) ligand 22 (CCL22) plays an important role in suppressing immune responses against H. pylori and tumor cells. In this study, the authors examined the association between single nucleotide polymorphisms (SNPs) in the CCL22 gene and the risk of gastric carcinoma. METHODS: Information on SNPs in the CCL22 coding region was obtained from the HapMap Project database. Genotypes were determined in a case-control cohort that consisted of 1001 patients with gastric carcinoma and 1066 controls, and odds ratios (ORs) and 95% confidence intervals (95% CIs) were computed by using a logistic regression model. Serum H. pylori antibody levels were measured by using an enzyme-linked immunosorbent assay. RESULTS: The 16C-->A SNP (reference SNP no. 4359426) in exon 1 of the CCL22 gene, which causes a 2 aspartate (2Asp) to 2 alanine (2Ala) substitution in the CCL22 protein, was associated with a significantly increased risk of gastric carcinoma. Individuals who were homozygous for the Ala/Ala genotype had an OR of 2.27 (95% CI, 1.28-4.02) compared with individuals who had the Asp/Asp genotype. Stratification analysis indicated that the association was more pronounced among men (OR, 2.64; 95% CI, 1.29-5.41) and among younger individuals (OR, 2.85; 95% CI, 1.36-5.96) compared with women and older individuals. Moreover, a multiplicative joint effect between the CCL22 SNP and H. pylori infection that intensified the risk was observed (OR for the presence of both Ala/Ala genotype and H. pylori infection, 18.37; 95% CI, 2.30-146.67). CONCLUSIONS: The results from this study suggested that the CCL22 polymorphism is associated with an increase risk of developing H. pylori infection-related gastric carcinoma. PMID- 19298003 TI - Interactions between metabolic and reproductive functions in the resumption of postpartum fecundity. AB - Lactation has long been recognized as a major determinant of interbirth intervals. The temporal pattern of nursing has been proposed as the mechanism behind lactational amenorrhea. We present a new model of the dynamic regulation of lactational amenorrhea that identifies maternal energy availability as the main determinant of ovarian resumption. Variation in the intensity of lactation remains a component of the model as a determinant of the absolute energetic cost of milk production. However, maternal energy supply determines net energy availability; a larger energy supply leaves a greater net energy surplus than a smaller energy supply (lactation costs being equal). We characterize the hormonal postpartum profile of 70 lactating Toba women of Argentina. We use C-peptide, which reflects maternal insulin production, as a measure of energy availability. Initially low, insulin production rises as the postpartum period progresses, reflecting the declining metabolic load of lactation. A short period of supernormal insulin production precedes menstrual resumption. The high levels of insulin may play a role in stimulating the resumption of ovarian activity, which in turn may help to resolve the transient period of insulin resistance. The dynamics of insulin sensitivity during lactation would aid in synchronizing the resumption of ovarian function with a reduction in the energy demands of milk production. This hypothesis is supported by the sustained weight gain experienced by lactating women during the months preceding the first postpartum menses. The link between fecundity and energy balance could serve as a mechanism for adjusting the duration of lactational amenorrhea to the relative metabolic load of lactation. PMID- 19298004 TI - Is there a trade-off between fertility and longevity? A comparative study of women from three large historical databases accounting for mortality selection. AB - Frontier populations provide exceptional opportunities to test the hypothesis of a trade-off between fertility and longevity. In such populations, mechanisms favoring reproduction usually find fertile ground, and if these mechanisms reduce longevity, demographers should observe higher postreproductive mortality among highly fertile women. We test this hypothesis using complete female reproductive histories from three large demographic databases: the Registre de la population du Quebec ancien (Universite de Montreal), which covers the first centuries of settlement in Quebec; the BALSAC database (Universite du Quebec a Chicoutimi), including comprehensive records for the Saguenay-Lac-St-Jean (SLSJ) in Quebec in the nineteenth and twentieth centuries; and the Utah Population Database (University of Utah), including all individuals who experienced a vital event on the Mormon Trail and their descendants. Together, the three samples allow for comparisons over time and space, and represent one of the largest set of natural fertility cohorts used to simultaneously assess reproduction and longevity. Using survival analyses, we found a negative influence of parity and a positive influence of age at last child on postreproductive survival in the three populations, as well as a significant interaction between these two variables. The effect sizes of all these parameters were remarkably similar in the three samples. However, we found little evidence that early fertility affects postreproductive survival. The use of Heckman's procedure assessing the impact of mortality selection during reproductive ages did not appreciably alter these results. We conclude our empirical investigation by discussing the advantages of comparative approaches. PMID- 19298005 TI - Grandparental investment: The influence of reproductive timing and family size. AB - The influence that grandparents have on the life history traits of their descendants has been studied extensively. However, no attention has been paid to the potential influence a grandparent's own reproductive history has on the investment they make in their grandchildren. We use data from 658 Swiss grandchildren and 591 of their grandparents to investigate whether grandparents' reproductive scheduling and family size influence the amount of investment grandparents make in a focal grandchild (shared contacts, occasions to meet, activities, discussions, interests, and important roles the grandparent plays). Grandparents who were younger when they had their first child had more children and grandchildren; this relationship strengthened after controlling for grandparental age, sex, lineage, and education (all P < 0.001). Generally, having more children or grandchildren was associated with reduced levels of grandparental investment. After adjustment for a wide range of factors known to influence investment, having more children or grandchildren and having a first child or grandchild at a younger age were associated with reduced investment in 14 of 24 analyses (all P < 0.09). The association between reproductive scheduling and investment was partially mediated by the grandparent's family size. Interestingly, these relationships were only present in data reported from the grandchild's point of view, not the grandparent's. This analysis provides preliminary evidence that grandparents' reproductive strategies have consequences for the amount of investment they make in their grandchildren. These results are examined in terms of the trade-offs between current and future reproduction and offspring quality and quantity. PMID- 19298006 TI - A comparison of physiological variables in aged and young women during and following submaximal exercise. AB - Previously, we have examined how aging affects the physiological responses of men to endurance exercise. In the present investigation, we aimed to extend our assessment of the influence of aging on exercise-induced responses by focusing on women. Ten young (20.3 +/- 0.3 years; mean +/- SE) and 10 aged (75.5 +/- 1.2 years) women performed 30 min of cycling at 60-65% of their predetermined peak oxygen uptake. Data for respiratory exchange ratio (RER), heart rate, blood pressure, rectal temperature, and plasma metabolites were collected before exercise, at the 15th and 30th min of exercise, and at 5 and 15 min postexercise. A two-way, repeated measures ANOVA with main effects of age and time was conducted on each variable. Our findings showed that age affected exercise induced responses of each variable quantified. Although RER, heart rate, temperature, and lactate were significantly (P < 0.05) higher among young women, blood pressure and glucose values were greater among aged women. Moreover, unlike previous results noted among men where age-related differences primarily occurred during postexercise recovery, in women the effect of aging was detected during exercise itself. The data presented here indicate that aging impacts physiological responses of women to prolonged endurance exercise even when relative intensity (% of peak oxygen uptake) is held constant. Combined with our earlier study on men, these findings suggest that gender interacts with aging to determine whether age-related differences are manifested during exercise itself, or during postexercise recovery. PMID- 19298007 TI - Analysis of fatty acids in early mid-life in fertile women: Implications for reproductive decline and other chronic health problems. AB - The relationship between adipose fatty acid levels and age is examined in 635 Scottish fertile women aged 25-48. Fatty acids levels are highly significantly correlated with age. Factor analysis identifies four factors that account for 79.6% of variance in the data. Three Factors show significant regressions with age and patterns of involvement of specific fatty acids suggest that these Factors represent the activity of fatty acid delta-desaturase enzymes as follows: Factor 1-delta-9-desaturase, Factor 2-delta-5-desaturase, and Factor 4-delta-6 desaturase. Key changes, apparently reductions in enzyme activity, occur through the 30 and 40-year-old age groups. Such changes in enzyme function could account for decline in female fertility and increases in body fat and chronic disease common in early mid-life. PMID- 19298008 TI - The intratumoral distribution of nuclear beta-catenin is a prognostic marker in colon cancer. AB - BACKGROUND: Most colon cancers harbor mutations of APC or beta-catenin, both of which may lead to nuclear beta-catenin accumulation in the tumor cells and constitutively activated expression of its target genes. In many colon cancers, however, nuclear beta-catenin accumulation is heterogeneous throughout the tumor and often confined to the tumor margin. Herein, the authors investigated whether the intratumoral distribution of nuclear beta-catenin can serve as a prognostic marker for survival and tumor progression of stage IIA colon cancer patients. METHODS: In total, 142 patients with primarily resected, moderately differentiated stage IIA colon cancer were included in this study. The patterning of nuclear beta-catenin expression was evaluated on immunohistochemically stained whole tissue sections of the tumors and was correlated with cancer-specific survival and disease-free survival using univariate and multivariate statistical analyses. RESULTS: Four distinct patterns of nuclear beta-catenin expression were identified, and 2 main categories comprising tumors with or without intratumoral regulation of nuclear beta-catenin were distinguished. Moreover, the results demonstrated that the patterning, and especially the regulation or absence of regulation of nuclear beta-catenin expression, was a strong predictive marker of patient survival and tumor progression. CONCLUSIONS: The current results indicated that the distribution of nuclear beta-catenin expression can be used as a good prognostic marker in patients with stage IIA colon cancer. Thus, the evaluation of nuclear beta-catenin may help to identify patients who will have a shorter than average survival and patients with a greater risk of disease progression who may be considered for adjuvant therapeutic modalities and intensified clinical aftercare in the future. PMID- 19298009 TI - Adults with acute lymphoblastic leukemia and translocation (1;19) abnormality have a favorable outcome with hyperfractionated cyclophosphamide, vincristine, doxorubicin, and dexamethasone alternating with methotrexate and high-dose cytarabine chemotherapy. AB - BACKGROUND: Among the well described cytogenetic abnormalities in adults with acute lymphoblastic leukemia (ALL), a translocation involving chromosomes 1 and 19 (t[1;19] [q23;p13]) occurs in a small subset but has been associated variously with an intermediate prognosis or a bad prognosis in different studies. METHODS: Adults with ALL and t(1;19) who were treated at The University of Texas M. D. Anderson Cancer Center were reviewed. Their clinical features and outcomes were compared with those of patients who had other cytogenetic abnormalities. The study endpoints included the complete remission (CR) rate, the complete response duration (CRD), and overall survival (OS). RESULTS: Of 411 adults with pre-B-cell ALL, 12 patients had t(1;19). Ten of 12 patients with t(1;19) received hyperfractionated cyclophosphamide, vincristine, doxorubicin (Adriamycin), and dexamethasone alternating with methotrexate and high-dose cytarabine (hyper-CVAD) chemotherapy; and the other 2 patients received combined vincristine, doxorubicin, and dexamethasone (VAD). All 12 patients achieved CR, and the 3-year survival rate was 73%. Patients with t(1;19) had significantly better CRD and OS compared with all other patients combined and compared individually with patients who had Philadelphia chromosome-positive, t(4;11), and lymphoma-like abnormalities (deletion 6q, addition q14q, t[11;14], and t[14;18]). CONCLUSIONS: Adults with ALL and t(1;19) had an excellent prognosis when the received the hyper-CVAD regimen. PMID- 19298010 TI - Growth perturbations in a phenotype with rapid fetal growth preceding preterm labor and term birth. AB - The variability in fetal growth rates and gestation duration in humans is not well understood. Of interest are women presenting with an episode of preterm labor and subsequently delivering a term neonate, who is small relative to peers of similar gestational age. To further understand these relationships, fetal growth patterns predating an episode of preterm labor were investigated. Retrospective analysis of fetal biometry assessed by serial ultrasound in a prospectively studied sample of pregnancies in Santiago, Chile, tested the hypothesis that fetal growth patterns among uncomplicated pregnancies (n = 3,706) and those with an episode of preterm labor followed by term delivery (n = 184) were identical across the time intervals 16-22 weeks, 22-28 weeks, and 28-34 weeks in a multilevel mixed-effects regression. The hypothesis was not supported. Fetal weight growth rate was faster from 16 weeks among pregnancies with an episode of preterm labor (P < 0.05), declined across midgestation (22-28 weeks, P < 0.05), and rebounded between 28 and 34 weeks (P = 0.06). This was associated with perturbations in abdominal circumference growth and proportionately larger biparietal diameter from 22 gestational weeks (P = 0.03), greater femur (P = 0.01), biparietal diameter (P = 0.001) and head circumference (P = 0.02) dimensions relative to abdominal circumference across midgestation (22-28 weeks), followed by proportionately smaller femur diaphyseal length (P = 0.02) and biparietal diameter (P = 0.03) subsequently. A distinctive rapid growth phenotype characterized fetal growth preceding an episode of preterm labor among this sample of term-delivered neonates. Perturbations in abdominal circumference growth and patterns of proportionality suggest an altered growth strategy pre dating the preterm labor episode. PMID- 19298011 TI - Finding and fostering the positive in relationships: positive interventions in couples therapy. AB - Research of positive psychology interventions (PPIs) has expanded dramatically in recent years, and many novel PPIs may be useful in couples therapy. The present work identifies, summarizes, and suggests adaptations of PPIs that may improve couples therapy outcomes. Each intervention is presented as part of a larger organizational framework that may help couples therapists determine how and when each intervention can be effectively applied. Finally, a case illustration demonstrates how these methods can complement traditional therapeutic approaches. PMID- 19298012 TI - Generation of transgenic mice expressing Cre recombinase under the control of the Dll1 mesoderm enhancer element. AB - To study paraxial mesoderm formation in the mouse, transgenic lines that can be used to either selectively delete or express genes of interest in the paraxial mesoderm are required. We have generated a transgenic mouse line that expresses Cre recombinase in the paraxial mesoderm (PAM) beginning at e7.5. A lacZ Cre recombinase reporter line showed that in addition to PAM and its derivatives, lateral plate and intermediate mesoderm derivatives were also exposed to Cre activity, while the node, notochord, and cardiac mesoderm were not. We further demonstrate that 70-75% of the fibroblasts generated from Dll1-msd Cre, ROSA26 rtTA embryos possess Cre recombinase activity. These mice can therefore be used in combination with tet-responsive transgenic lines to generate mesoderm-derived embryonic fibroblasts that inducibly express a gene of interest. PMID- 19298013 TI - Molecular cloning of a dominant roller mutant and establishment of DNA-mediated transformation in the nematode Pristionchus pacificus. AB - We report the molecular cloning of a dominant Roller mutant of Pristionchus pacificus, which encodes a cuticle collagen. We use the mutant locus as a marker to develop transgenic technique by generating complex arrays and present flourescent-protein based transcriptional reporter constructs for P. pacificus. PMID- 19298014 TI - Gonadal sex differentiation and expression of Sox9a2, Dmrt1, and Foxl2 in Oryzias luzonensis. AB - Oryzias luzonensis is closely related to the medaka, O. latipes. The sex of both species is determined by an XX-XY system. However, the testis determining gene (DMY/Dmrt1bY) found in O. latipes does not exist in O. luzonensis. Instead, a different gene is thought to act as a testis determining gene. In this study, we focused the gonadal sex differentiation process in O. luzonensis under different testis determining gene. First, we observed the gonadal development of O. luzonensis histologically. We then analyzed the expression of Sox9a2/Sox9b, Dmrt1, and Foxl2 during early development. Our results suggest that the sexual differentiation of germ cells in O. luzonensis is initiated later than in O. latipes. However, the timing of the sexual differentiation of the supporting cell linage is similar between the species. PMID- 19298015 TI - Lmx1b-expressing cells in the mouse limb bud define a dorsal mesenchymal lineage compartment. AB - The LIM-homeodomain transcription factor lmx1b is a critical regulator of vertebrate dorsal-ventral limb patterning. In the mouse embryo, lmx1b is initially transcribed throughout most, if not all, limb bud cells at early stages, but then rapidly becomes restricted specifically to dorsal mesenchymal cells with a sharp boundary between the dorsal-positive and ventral-negative expression domains. How this expression pattern is initially established is not well understood, nor are mechanism(s) that maintain a sharp dorsal-ventral boundary between lmx1b expressing and nonexpressing cells. Here, we employ a genetic fate mapping approach to establish that the transition from a broad expression domain of lmx1b to a restricted dorsal domain of expression involves selective loss of lmx1b expression in presumptive ventral cells. In addition, we show that once lmx1b expression becomes restricted to dorsal mesenchyme cells, these cells form a lineage-based compartment that prevents mixing between dorsal and ventral cells, consistent with recent fate mapping experiments carried out the chick and mouse (Pearse et al.,2007, Dev Biol 310:388-400; Arques et al.,2007, Development 134:3713-3722). Moreover, lmx1b activity is required to maintain, but not to establish the dorsal mesenchymal compartment likely through a mechanism involving differential cell adhesion. Taken together, our results indicate that lmx1b expressing cell define a dorsal limb bud mesenchymal lineage compartment and that maintenance of this compartment depends on lmx1b function. PMID- 19298016 TI - Case studies of ends-out gene targeting in Drosophila. AB - Ends-in and ends-out gene replacement approaches have been successfully used to disrupt Drosophila genes involved in a variety of biological processes. These methods combine double-strand breaks and homologous recombination to replace a targeted chromosome region with a designed DNA sequence. Unfortunately, these methods require large numbers of single animal crosses, making them both time consuming and labor intensive. Here, we designed a single complete targeting vector for use in a mass crossing ends-out gene targeting study. Importantly, our gene targeting method included a balancer chromosome to block endogenous homologous chromosome pairing and to promote pairing between the foreign targeting DNA fragment and the targeted chromosome. This technique provided successful and efficient gene replacement, greatly facilitating the gene knockout procedure. PMID- 19298017 TI - Systemic metal exposure in large- and small-diameter metal-on-metal total hip replacements. AB - Large-diameter metal-metal total hip replacement (THR) offers the advantages of low wear and low dislocation risk. the aim of this investigation was to compare metal levels in large and small-diameter metal-metal hip replacements. Whole blood concentrations and daily output of cobalt and chromium in 28 patients with unilateral large diameter (42- to 54-mm) metal on metal hip replacements at 1 year follow-up were compared with levels in patients with 28-mm metal-on-metal THRs. Both bearings were made of high-carbon cobalt-chrome alloy. The larger bearing is as-cast and the smaller wrought alloy. High-resolution inductively coupled plasma mass spectrometry was used for analysis. The patients had either a cemented polished tapered stainless steel stem or a cementless porous ingrowth titanium alloy stem. Mean whole blood levels in the small- and large-diameter THRs are not significantly different at 1 year (cobalt, 1.7 vs 2.3 microg/L and chromium 1.7 vs 1.4 microg/L). Daily urinary output of cobalt and chromium was also in the same range and without a significant mean difference (cobalt 11.6 microg/24 h in large-diameter and 12.3 microg/24 h in small-diameter THRs and chromium 3.7 and 4.1 microg/24 h, respectively). PMID- 19298018 TI - A novel blood-saving plan for less-invasive primary total hip replacement. AB - We conducted a quality improvement program to examine the effect of a blood saving plan during primary total hip replacement (THR) performed using a nonvascular less invasive approach (LI-THR) and compared the results with historical control subjects. Erythropoietin and tranexamic acid (TA) were administered for selected patients. Analysis of 221 (study group) and 186 (historic group) LI-THRs showed reductions in total blood loss by 20% in a group of 133 patients compared to a control group of 82 patients who did not receive TA. This novel blood plan for LI-THR changed practice, improved quality of care, and allowed all patients to remain blood transfusion free. PMID- 19298019 TI - Safe and accurate: learning the direct anterior total hip arthroplasty. AB - Wear, instability, leg length, and muscle recovery are the major obstacles in total hip arthroplasty (THA). The direct anterior approach with fluoroscopic assistance has been proposed to address all four of these issues. The goal of this study was to assess the learning curve, safety, and accuracy of direct anterior THA. A retrospective review was completed on a single surgeon's initial consecutive series of 300 THAs performed via a direct anterior approach. Cases were grouped based on chronologic order (1-100, 101-200, 201-300). Operative time, fluoroscopy time, estimated blood loss, radiographic leg length discrepancy (LLD), radiographic cup abduction angle, and complications were recorded. Data were analyzed using an analysis of variance. Postoperatively, all patients were managed without dislocation precautions. The average age for the cohort was 58.9 years, and the average BMI was 29. All groups were similar with respect to age and BMI. Direct anterior THA demonstrated significant reductions inoperative and fluoroscopy after the first 100 cases. Mean surgery time was 132.8 minutes for group 1, 109.9 for group 2, and 106.1 for group 3 (P<.001). Mean fluoroscopy time was 32.1 seconds for group 1, 14.5 for group 2, and 14.5 for group 3 (P<.001). There was one dislocation in group 3 and three intraoperative calcar fractures in the first 100 cases. There were no infections. Direct anterior THA demonstrated a reduction in operative time and fluoroscopy time after the initial 100 cases. Calcar fracture did not occur after the first 62 cases. Cup abduction angle, dislocation rate, and LLD were excellent in all groups. PMID- 19298020 TI - A clinicoradiologic study of the Birmingham Mid-Head Resection device. AB - This is a 1.2- to 5.3-year survival and clinicoradiologic study of patients with the Birmingham Mid-Head Resection (BMHR) device (Smith & Nephew Orthopaedics, Warwick, United Kingdom). Sixty consecutive hips implanted with this device between 2003 and 2007 were reviewed with hip outcome questionnaires, clinical examination, and radiographs at a minimum follow-up of 1 year. There were no revisions, mechanical failures, or signs of femoral neck stress shielding. All hips were functioning well and showed no significant adverse clinical or radiographic features. Birmingham Mid-Head Replacement offers the prospect of circumventing the need for a more invasive procedure such as hip replacement in patients who would benefit from a conservative arthroplasty but lack femoral head bone quality, which is a prerequisite for a successful hip resurfacing. PMID- 19298021 TI - When is the right time to resurface? AB - With the recent approval of 2 metal-on-metal hip resurfacing devices in the United States, hip resurfacing is being performed more commonly. As with most orthopedic procedures, appropriate indications are the key to successful outcomes and avoiding complications such as femoral neck fracture. A review of the literature suggests that optimal results, and the lower risk of early failure, are obtained in men with osteoarthritis who are younger than age 55 years. This article reviews general considerations for choosing appropriate candidates for metal-on-metal hip resurfacing, including relative and suggested contraindications to the procedure. PMID- 19298022 TI - Early clinical experience with the use of the Birmingham hip resurfacing system. AB - This study reviews the early clinical experience with the Birmingham Hip resurfacing system (Smith & Nephew, Memphis, Tenn) in the United States since its approval by the FDA. A total of 230 patients were followed for a mean of 16 months (range, 6 months). There was a significant improvement in the mean preoperative Oxford hip score at the latest follow-up (44 points [range, 30-58 points] to 17 points [range, 12-28 points]). The most commonly used femoral component was 50 mm, and the mean duration of the procedures was 95 minutes (range, 65-180 min). there were 3 revisions (1.3%). Early results are favorable. As with any device, there is a continuing need for longterm monitoring and large scale epidemiologic surveillance. PMID- 19298023 TI - Comparison of mid-term clinical outcomes after primary total hip arthroplasty with Oxinium vs cobalt chrome femoral heads. AB - The purpose of this investigation was to assess the midterm clinical outcomes after implantation of Oxinium (OX) vs cobalt-chrome (CC) femoral heads. Primary total hip arthroplasty (THA) procedures were performed in 100 patients. After randomization, half of the patients received OX femoral heads and half received CC femoral heads. At a minimum follow-up of 2-years, stem survival was 98% for both groups. The mean Harris Hip score was 92 for OX and 92.5 for CC, with mean WOMAC scores of 84.9 and 87, respectively. For SF-12, the OX group had mean physical and mental component scores of 45.2 and 53.8 and the CC group 49.2 and 52.6, respectively. At a minimum follow-up of 2 years, clinical outcomes for THA procedures using OX and CC femoral heads appear equivalent. PMID- 19298024 TI - Emperion noncemented modular femoral component--can modularity solve any primary total hip replacement problem? AB - The Emperion femoral component (Smith & Nephew, Memphis, Tenn) is a modern incarnation of the S-ROM femoral component (Depuy, Warsaw, Ind). This fit-and fill modular design has had great clinical success for more than 20 years. The Emperion can be used to address almost all clinical scenarios in primary total hip replacement (THR). The modularity can uniquely weather difficult proximal femoral deformities encountered during THR, including developmental dysplasia of the hip and slipped capital femoral epiphysis. In addition, the nomenclature attached to this implant has made using the device and its instruments user friendly. PMID- 19298025 TI - Echelon stems and primary total hip replacement surgery. AB - The Echelon Primary femoral stem is an extensively porous coated cylindrical cobalt chrome hip component. A prospective review of 392 Echelon stems revealed excellent survivorship of the stem, with a 99.3% survival rate for aseptic loosening and a 98.3% survival rate for revision for any reason at 8 years. Normalized Western Ontario and McMaster Universities Osteoarthritis Index and short-form health survey outcome scores were significantly improved and subsequently maintained after replacement, with 82% of patients obtaining a good or excellent result at the mean follow-up period. There have been no changes in manufacture or design of the stem during the review period. PMID- 19298026 TI - The dual-mobility POLARCUP: first results from a multicenter study. AB - Dislocation is a leading cause of revision after total hip arthroplasty (THA). To address this risk, dual-mobility technology was developed, which features a mobile polyethylene liner locked onto a femoral head and articulating in a metallic acetabular shell. This study reports clinical outcome data after implantation of the third-generation POLARCUP Dual-Mobility System (Smith & Nephew Orthopaedics AG, Rotkreuz, Switzerland). Primary THA procedures were performed in 150 patients. At 7.1 years, cumulative cup survival according to Kaplan-Meier was 97.4%. The mean Postel-Merle d'Aubigne score improved from 8.9 to 17.1 during the investigation. Two cups were revised at 5.4 and 6.4 years because of aseptic loosening. No dislocations were observed during follow-up. The current results confirm excellent early to midterm clinical outcomes for the POLARCUP Dual-Mobility System. PMID- 19298027 TI - The next generation of acetabular shell design and bearing surfaces. AB - The R3 Acetabular component represents the next generation of acetabular shell with an enhanced porous ingrowth surface (StikTite, Smith & Nephew, Memphis, Tenn) to meet the needs of both primary and revision hip arthroplasty; an optimized locking mechanism; and the ability to accomodate polyethylene, metal, or ceramic liners. This prospective clinical study reports on the safety and efficacy of the new StikTite porous ingrowth surface using radiostereometric analysis (RSA). StikTite provides a superior "sctratch-fit" due to its greater coefficient of friction and less micromotion using RSA measurements. PMID- 19298028 TI - Review of the evolution of the cementless acetabular cup. AB - First-generation porous coated acetabular cups have achieved durable biologic fixation, although liner dissociation and wear were common failure mechanisms. Second-generation modular acetabular cups retained the successful ingrowth surfaces used in earlier cup designs and incorporated congruent contact between the liner and shell. However, impingement-related fractures of highly cross linked ultra-high-molecular-weight-polyethylene (UHMWPE) used in second generation cup designs have occurred. Third-generation cup designs that eliminate UHMWPE protruding above the metal rim and minimize sharp corners at the liner locking mechanism would be expected to reduce the risk of liner fracture in modular acetabular cups used with highly crosslinked UHMWPE. PMID- 19298029 TI - Validation of computer-assisted open-wedge high tibial osteotomy using three dimensional navigation. AB - An unintended increase in the posterior tibial slope after open-wedge high tibial osteotomy (HTO) can influence knee kinematics and stability. The objective for this study was to validate the change of the tibial slope obtained from three dimensional (3D) navigation in open-wedge HTO by comparing it with that evaluated with computed tomography. Human cadaver knees were used. The open-wedge HTO was performed to maintain the anatomic tibial slope according to the navigation system. 3D navigation could provide surgeons with reliable information not only to determine appropriate coronal alignment but also to maintain the anatomic tibial slope in open-wedge HTO. PMID- 19298030 TI - Accuracy of navigation: a comparative study of infrared optical and electromagnetic navigation. AB - We evaluated the accuracy of navigation systems for measuring the mechanical axis in patients undergoing total knee arthroplasty and in the synthetic bone model. Infrared optical and electromagnetic navigation systems were compared. Both systems were found to be accurate and reproducible in an experimental environment. However, the accuracies of both systems were affected by erroneous registration, and the optical system was found to be more reproducible. In clinical situations, the mean difference was 1.23 degrees, and difference greater than 3 degrees occurred in 15% of clinical trials. These discordances may have been due to ambiguous anatomic landmarks causing registration errors and the possibility of electromagnetic signal interference in the operating room. PMID- 19298031 TI - Real-time computer-assisted notch assessment in anterior cruciate ligament repair. AB - Graft impingement can cause graft failure or movement restrictions. Intraoperative computer-assisted referencing of the tibial and femoral geometry, including precise mapping of the intercondylar notch, allows placement of the tunnels by real-time calculation balancing the impingement risks vs isometry. Biological fixation with bone cylinders locks the graft flush with the joint line and requires more accurate tunnel placement to avoid graft impingement. In 45 patients who underwent such navigation controlled tunnel placement 41 had no need for notchplasty. In two cases, the notchplasty was performed immediately for obvious osteophyte restriction. A notchplasty was added two times after the first referencing to obtain satisfactory isometry. At staged follow up to 18 months, we found no laxity, flexion contracture with a mean flexion arc over 130 degrees, or tunnel widening. With computer-navigated realtime assessment of notch geometry, full functional recovery and stability were obtained, and unnecessary notchplasties were avoided even with a less flexible biological graft fixation. PMID- 19298032 TI - Analysis of the patellofemoral congruence angle according to the rotational alignment of the femoral component in navigation-guided TKA. AB - The relative femoral resection plane from the posterior condylar axis was determined by the navigation system. The investigators found that there was a relatively variable range of femoral component rotation intraoperatively (0 degrees - 6 degrees) and attempted to determine whether this would affect postoperative patellofemoral congruence. Forty-six varus knees from 34 patients were included in the study; group 1 (15 knees) with 0 degrees or 1 degrees and group 2 (31 knees) with 3 degrees to 6 degrees . The mean (P = .855) and percentage of abnormal values (patellofemoral congruence angle <16 degrees) (P = .193) in preoperative radiographs showed no significant differences between the two groups. In postoperative findings, the mean of patellofemoral congruence angles in group 1 (20.5 degrees) showed a higher tendency than that in group 2 (14.1 degrees), but no statistically significant difference between two groups (P = .089). In conclusion, there was no statistically significant difference in patellofemoral congruence between 2 groups. PMID- 19298033 TI - Unicompartmental knee replacement: a comparison of four techniques combining less invasive approach and navigation. AB - We developed a nonimage-guided navigation system for unicompartmental knee replacement, suitable for both conventional and minimally invasive approaches. We performed a radiologic analysis of the accuracy of implantation with conventional nonnavigated instrumentation, conventional, open navigated instrumentation, minimally invasive navigated experimental instrumentation derived from conventional instruments, and minimally invasive navigation-dedicated instrumentation. Navigated technique allowed improving the accuracy of the radiologic implantation. Minimally invasive implantation was effective, but the accuracy may not reach that of the conventional navigated technique. Minimal invasive techniques have to be validated, because a loss of accuracy will negatively influence longterm outcomes. PMID- 19298034 TI - Columbus primary total knee replacement: a 2- to 4-year followup of the use of intraoperative navigation-derived data to predict pre and postoperative function. AB - The purpose of this study was to examine the clinical outcomes associated with the Columbus primary total knee replacement (B. Braun Aesculap, Tuttlingen, Germany), correlate these outcomes with variables measured intraoperatively with the OrthoPilot navigation system (B. Braun Aesculap), and explore the full potential of automating the process of intraoperative data collection. Clinical and functional outcomes at 2.5 years were similar to results reported in previous studies. Correlations were seen between initial mechanical axis deformity and postoperative range of motion as well as between final mechanical axis alignment and the presence of flexion contractures at later followup. It is now possible to potentially stratify particular segments of patients and develop specific intraoperative alignment targets that are most likely to yield positive clinical and functional outcomes. PMID- 19298035 TI - One-year followup of 214 total knee arthroplasties with navigated columbus implants. AB - In this study, 206 patients with 214 Columbus total knee arthroplasty (TKA) implants were followed up at 1 year. Preoperatively, patient demographics (mean [SD]) were 85 male; age, 69.7 (8.7) years; ASA score, 2.5 (0.7); body mass index, 32.2 (5.7); 161 degrees varus and 27 degrees valgus; fixed flexion, 5.6 degrees (6.1 degrees); flexion, 96.1 degrees (18.8 degrees); and Oxford score, 43 (7.0). At 1-year follow-up, results were fixed flexion, 0.9 degrees (2.6): maximum, 17 degrees, minimum, 0 degrees; flexion, 101.3 degrees (9.1): maximum, 125 degrees, minimum, 75 degrees; and Oxford score, 23 (7.7). Radiographs showed radiolucent lines in 6 femurs in 1 zone; 1 in 2 zones and 0 in more than 2 zones; and 3 tibias in 1 zone. There were 2 deep infections. Ninety-eight percent of patients were satisfied with their TKA. PMID- 19298036 TI - The reliability of navigation-guided gap technique in total knee arthroplasty. AB - The OrthoPilot TKA navigation system (B. Braun Aesculap, Tuttlingen, Germany) offers software to optimize soft tissue balance using gap balance techniques. However, there are no studies on the reliability of the navigation-guided gap technique. The goal of this study is to establish the reliability of the navigation-guided gap technique. The investigators measured flexion and extension gap in the medial and lateral sides of the knee joint after bone resection to evaluate the reliability of navigation-guided soft tissue balancing. Gap data from 100 cases of navigation-guided total knee arthroplasty were analyzed. We defined trapezoidal gap (unsatisfactory soft tissue balance) as a gap difference < 3 mm between the medial and lateral sides in extension and a 5 mm difference in 90 degrees of flexion. Furthermore, gap difference between flexion and extension greater than 3 mm on the medial side and 5 mm on the lateral side was also considered a trapezoidal gap. Among 100 cases, 84 showed rectangular (acceptable) gap, and 16 showed trapezoidal gap. We also evaluated the correlation between clinical results including range of motion and soft tissue balance as well as characteristics of trapezoidal gap. This study suggests that the navigationguided gap technique is a reliable method for optimizing soft tissue balance. PMID- 19298037 TI - Leg length discrepancy, dislocation rate, and offset in total hip replacement using a short modular stem: navigation vs conventional freehand. AB - We present a match-paired study between computer-assisted and freehand techniques using a short modular femoral stem (Metha; B. Braun Aesculap, Tuttlingen, Germany) in total hip replacement (THR). Surgical time, clinical outcome, dislocation rate, limb length, and offset in 44 patients with ideal indication for this more conservative implant were assessed. Despite both longer surgical time and similar early outcomes, the results showed how computer-assisted techniques allow easier management of limb length discrepancy and offset restoring. We believe that navigated short modular stems are safe for less invasive THR. PMID- 19298038 TI - Comparison between pointer-based and ultrasound-based navigation technique in THA using a minimally invasive approach. AB - The use of navigation techniques in primary total hip arthroplasty improves the position of endoprosthetic components, especially cup positioning. An intraoperative registration of the anterior pelvic plane is necessary to define the anteversion and inclination angles on the acetabular side. This study compares the accuracy of manual pointer palpation to ultrasound registration in navigation to determine pelvic plane registration in 60 cases of minimally invasive surgical technique. Findings show more accurate postoperative radiographic anteversion with ultrasound navigation, although both manual pointer palpation and ultrasound registration techniques show a very small standard deviation in anteversion, inclination, and leg length difference. In conclusion, we recommend navigation as a very reliable tool for the positioning of implants. PMID- 19298039 TI - Comparative study of acetabular center axis vs anterior pelvic plane registration technique in navigated hip arthroplasty. AB - There is significant variation in registering the anterior pelvic plane (APP) among experienced navigated hip arthroplasty surgeons, reflecting negatively on the accuracy of determining inclination and anteversion. Whether it is variation in pelvic anatomy or improper positioning, this inaccuracy emphasizes the need for alternative methods of registration, of which the acetabular center axis (ACA) is proposed. Data collected from ACA and APP registration were compared with postoperative computed tomography (CT) images of the pelvis in 34 cases. Findings showed ACA software to be comparable with CT in its accuracy in determining the inclination and version angles of the acetabulum and cup implant. PMID- 19298040 TI - Navigated and nonnavigated total hip arthroplasty: results of two consecutive series using a cementless straight hip stem. AB - The purpose of this study was to compare conventional and navigated technique and a recently developed straight hip stem for uncemented primary total hip arthroplasty. The results of two consecutive implantation series of 42 patients (nonnavigated) and 42 patients (navigated) were analysed for implant positioning and complications. All surgeries were performed by the investigator. Radiographic analysis of cup position showed a significant improvement with reduced radiologic inclination (53 degrees nonnavigated /44 degrees navigated; P < .001) and higher anteversion (7 degrees nonnavigated /12 degrees navigated; P <.001). The mean postoperative limb length difference was 6.2 mm (SD, 9.0 for nonnavigated) and 4.4 mm (SD, 6.4 for navigated). Intraoperative and early postoperative complications were not different. No dislocation occurred in either group. There was one intraoperative trochanter fracture that was not revised (nonnavigated) and one revision because of a periprosthetic fracture caused by fall down during rehabilitation (navigated). We conclude that acetabular implant positioning can be significantly improved by the use of navigated surgery technique. The data for postoperative limb length difference were still similar but within the expected range in both groups. The effect of improved cup positioning on mid and longterm results for both groups has to be investigated further. PMID- 19298041 TI - C-D modes of deuterated side chain of leucine as structural reporters via dual frequency two-dimensional infrared spectroscopy. AB - Perdeuteration of the side chains of amino acids such as leucine results in appearance of reasonably strong absorption peaks around 2050-2220 cm(-1) that belong to the CD stretching modes and exhibit extinction coefficients of up to 120 M(-1) cm(-1). The properties of the CD stretching transitions in leucine d(10) as structural labels are studied via the methods of two-dimensional infrared (2DIR) spectroscopy. The cross peaks caused by interactions of the CD stretching modes with amide I (Am-I), CO, and amide II (Am-II) modes are obtained by the dual-frequency 2DIR method. The CD stretching peaks in leucine-d(10) are characterized using DFT computational modeling as well as relaxation-assisted 2DIR (RA 2DIR) measurements. The RA 2DIR measurements showed different enhancements and different energy transport times (arrival times) for different CD/Am-II and CD/CO cross peaks; a correlation between the intermode distance, the arrival time, and the amplification factor is reported. We demonstrated that the CD transitions of leucine-d(10) amino acid can serve as convenient structural reporters via the dual-frequency 2DIR methods and discussed the potential of leucine-d(10) and other amino acids with deuterium-labeled side chains as probes of protein structure and dynamics. PMID- 19298042 TI - Exploring complex protein-ligand recognition mechanisms with coarse metadynamics. AB - The metadynamics method has been shown to be a valuable tool to study the mechanism of molecular recognition in atomistic detail [Gervasio, F. L.; et al. J. Am. Chem. Soc. 2005, 127, 2600]. However, it requires an a priori knowledge of all slow degrees of freedom relevant to the docking/undocking mechanism. Here we investigate a combination of docking/clustering with metadynamics performed with a subset of the necessary degrees of freedom (coarse metadynamics), and show that it provides a full mechanistic insight on the protein-ligand docking mechanism. Moreover, the proposed protocol is able to clearly distinguish between crystallographic and noncrystallographic poses of protein-ligand complexes, and also to find the transition state of the full undocking mechanism, thus giving an indication on the binding free energy. PMID- 19298043 TI - Production of a bioactive peptide (IIAEK) in Escherichia coli using soybean proglycinin A1ab1b as a carrier. AB - To produce large amounts of a peptide of fewer than 10 amino acid residues, construction of a gene encoding multimers of the small peptide is necessary. For this study a method was developed to facilitate the gene construction of high multimers of a small peptide with one step of cloning. A hypocholesterolemic peptide, IIAEK, from cow's milk beta-lactoglobulin was used as a model peptide for the construction of a gene encoding multimers of IIAEK and for the production of the peptide. Two systems for direct expression of 28-mers of IIAEK sequences (28IIAEK) and expression of 34 IIAEK sequences (4 IIAEK sequences in each of the disordered regions I, II, and III and 14 and 8 IIAEK sequences in disordered regions IV and V, respectively) in a mutant of soybean proglycinin A1aB1b lacking 31 residues in disordered region IV [A1aB1b(Delta31)-34IIAEK] were used. The protein produced from both systems formed inclusion bodies. The expression level of A1aB1b(Delta31)-34IIAEK was 29.9% of the total cell proteins and that of the 28IIAEK was 2.0%. The insoluble A1aB1b(Delta31)-34IIAEK was digested by trypsin without any help from urea or chemicals, and the produced IIAEK was purified using an octadecyl silica column. The yield of IIAEK was 58.6%. The results showed that A1aB1b as a carrier of multiple peptides and use of an Escherichia coli expression system are suitable for production of bioactive peptide. PMID- 19298044 TI - Identification of novel orosensory active molecules in cured vanilla Beans (Vanilla planifolia). AB - Sequential application of solvent extraction, gel permeation chromatography, and HPLC in combination with taste dilution analyses, followed by LC-MS and 1D/2D NMR experiments, led to the discovery of seven velvety mouth-coating molecules in cured beans of Vanilla planifolia . Among these, 5-(4-hydroxybenzyl)vanillin, 4 (4-hydroxybenzyl)-2-methoxyphenol, 4-hydroxy-3-(4-hydroxy-3-methoxybenzyl)-5 methoxybenzaldehyde, (1-O-vanilloyl)-(6-O-feruloyl)-beta-d-glucopyranoside, americanin A, and 4',6'-dihydroxy-3',5-dimethoxy-[1,1'-biphenyl]-3-carboxaldehyde were previously not reported in vanilla beans. Sensory studies revealed human recognition thresholds for the velvety mouth-coating sensation between 1.0 and 5.0 mumol/kg (water). Interestingly, the biphenyl derivatives were found to enhance the perception of creaminess and fatty body of sweetened skim milk, among which 4',6'-dihydroxy-3',5-dimethoxy-[1,1'-biphenyl]-3-carboxaldehyde showed the lowest threshold level of 5 mumol/kg. Quantitative analysis of these compounds in cured vanilla beans from different origins as well as in noncured beans revealed that, with the exception of americanin A, all of the other taste compounds are not present in the green vanilla beans and are formed during the bean curing process. PMID- 19298045 TI - Quantum wave packet method for state-to-state reactive scattering calculations on AB + CD --> ABC + D reactions. AB - We describe a quantum wave packet method for computing the state-to-state quantum dynamics of 4-atom AB + CD --> ABC + D reactions. The approach is an extension to 4-atom reactions of a version of the reactant-product decoupling (RPD) approach, applied previously to 3-atom reactions ( J. Chem. Phys. 2001, 114 , 1601 ). The approach partitions the coordinate space of the reaction into separate reagent, strong-interaction, and product regions, using a system of artificial absorbing and reflecting potentials. It employs a partitioned version of the split-operator propagator, which is more efficient than partitioning the (exact) time-dependent Schrodinger equation. The wave packet bounces off a reflecting potential in the entrance channel, which generates a source term; this is transformed efficiently from reagent to product Jacobi coordinates by exploiting some simple angular momentum properties. The efficiency and accuracy of the method is demonstrated by numerical tests on the benchmark OH + H(2) --> H(2)O + H reaction. PMID- 19298046 TI - Organometallic complexes for nonlinear optics. 43. Quadratic optical nonlinearities of dipolar alkynylruthenium complexes with phenyleneethynylene/phenylenevinylene bridges. AB - The syntheses of trans-[Ru(4,4'-C[triple bond]CC(6)H(4)C[triple bond]CC(6)H(4)NO(2))Cl(dppe)(2)] (19) and the systematically varied complexes trans-[Ru(4,4',4''-C[triple bond]CC(6)H(4)X(2)C(6)H(4)Y(2)C(6)H(4)NO(2))Cl(L(2))(2)] [L(2) = dppe, X(2) = C[triple bond]C, Y(2) = (E)-CH=CH (12), C[triple bond]C (18); L(2) = dppe, X(2) = (E)-CH=CH, Y(2) = C[triple bond]C (14), (E)-CH=CH (16); L(2) = dppm, X(2) = C[triple bond]C, Y(2) = (E)-CH=CH (13); L(2) = dppm, X(2) = (E)-CH=CH, Y(2) = C[triple bond]C (15), (E)-CH=CH (17)] are reported, the latter being donor-bridge acceptor complexes varying in bridge composition by replacement of yne with E-ene linkages, together with their cyclic voltammetric data, linear optical, and quadratic nonlinear optical response data. Ru(II/III) oxidation potentials increase on replacing yne linkage by E-ene linkage at the phenylene adjacent to the metal center, and on replacing dppe by dppm co-ligands. The low-energy optical absorption maxima occur in the region 20,400-23,300 cm(-1) and are metal to-ligand charge-transfer (MLCT) in origin; these bands undergo a blue-shift upon pi-bridge lengthening by addition of phenyleneethynylene units, and on replacing E-ene linkages by yne linkages. Time-dependent density functional theory calculations on model complexes have suggested assignments for the low-energy bands. The optical spectra of selected oxidized species contain low-energy ligand to-metal charge transfer (LMCT) bands centered in the region 9760-11,800 cm(-1). Quadratic molecular nonlinearities from hyper-Rayleigh scattering (HRS) studies at 1064 nm reveal an increase in the two-level-corrected beta(0) value on pi bridge lengthening, a trend that is not seen with beta values because of the blue shift in lambda(max) for this structural modification. Replacing yne linkages by E-ene linkage at the phenylene adjacent to the metal center or dppm co-ligand by dppe results in an increase in beta and beta(0) values. In contrast, quadratic molecular nonlinearities by HRS at 1300 nm or electric field-induced second harmonic generation (EFISH) studies at 1907 nm do not afford clear trends. PMID- 19298047 TI - Spectroscopic study of the ion-radical H-bond in H4O2+. AB - The primary event in the ionization of water involves rapid proton transfer, leading to charge localization on H(3)O(+) and the creation of a hydroxyl radical. We trap the nascent [H(3)O(+).(*)OH] exit channel intermediate in the bimolecular reaction by Ar-mediated ionization of the neutral water dimer and characterize the nature of this ion-radical complex using vibrational predissociation spectroscopy of the Ar-tagged species. The resulting bands involving the displacement of the bridging proton are broad and appear as a strong triplet centered around 2000 cm(-1). The observed band pattern is analyzed with theoretical calculations to identify the origin of the anhamonic effects evident in the spectrum. In the course of this work, expressions were derived for treating the coupling terms within a sinc-DVR. Although this level of treatment did not reveal the assignment of the triplet structure, its characteristic approximately 100 cm(-1) spacing suggests activity involving the frustrated rotation of the hydroxyl radical upon excitation of the bridging-proton vibration parallel to the heavy atom axis. The behavior of this system is considered in the context of that reported previously for the related H(5)O(2)(+), H(3)O(2)(-), and F(-).H(2)O complexes. PMID- 19298048 TI - A tale of tails: how histone tails mediate chromatin compaction in different salt and linker histone environments. AB - To elucidate the role of the histone tails in chromatin compaction and in higher order folding of chromatin under physiological conditions, we extend a mesoscale model of chromatin (Arya, Zhang, and Schlick. Biophys. J. 2006, 91, 133; Arya and Schlick. Proc. Natl. Acad. Sci. U.S.A. 2006, 103, 16236) to account for divalent cations (Mg(2+)) and linker histones. Configurations of 24-nucleosome oligonucleosomes in different salt environments and in the presence and absence of linker histones are sampled by a mixture of local and global Monte Carlo methods. Analyses of the resulting ensembles reveal a dynamic synergism between the histone tails, linker histones, and ions in forming compact higher-order structures of chromatin. In the presence of monovalent salt alone, oligonucleosomes remain relatively unfolded, and the histone tails do not mediate many internucleosomal interactions. Upon the addition of linker histones and divalent cations, the oligonucleosomes undergo a significant compaction triggered by a dramatic increase in the internucleosomal interactions mediated by the histone tails, formation of a rigid linker DNA "stem" around the linker histones' C-terminal domains, and reduction in the electrostatic repulsion between linker DNAs via sharp bending in some linker DNAs caused by the divalent cations. Among all histone tails, the H4 tails mediate the most internucleosomal interactions, consistent with experimental observations, followed by the H3, H2A, and H2B tails in decreasing order. Apart from mediating internucleosomal interactions, the H3 tails also contribute to chromatin compaction by attaching to the entering and exiting linker DNA to screen electrotatic repulsion among the linker DNAs. This tendency of the H3 tails to attach to linker DNA, however, decreases significantly upon the addition of linker histones due to competition effects. The H2A and H2B tails do not mediate significant internucleosomal interactions but are important for mediating fiber/fiber intractions, especially in relatively unfolded chromatin in monovalent salt environments. PMID- 19298049 TI - Evidence for initiation of thermal reactions of alkenes with hydrogen-terminated silicon by surface-catalyzed thermal decomposition of the reactant. AB - New insights into the mechanism of thermal reactions of alkenes with hydrogen terminated silicon are presented. Scanning tunneling microscopy (STM) imaging at the early stages of the reaction of 1-decene with H/Si(111) at 150 degrees C confirm this reaction occurs via a propagating radical chain mechanism. In addition, evidence is presented for an initiation mechanism involving degradation of hydrocarbon molecules catalyzed by the silanol surface of Schlenk tubes commonly used in carrying out these reactions. Hydrogen-terminated silicon surfaces are found to be unstable in the "inert" solvent dodecane when heated at 150 degrees C in a Pyrex Schlenk tube. By contrast, the surfaces were significantly more stable at the same temperature when reactions were carried out in Teflon (polytetrafluoroethylene or PTFE). The thermal reaction of decene with H/Si(111) was found to proceed more rapidly in Pyrex than in PTFE, consistent with an impurity-based initiation mechanism. PMID- 19298050 TI - The importance of being tyrosine: lessons in molecular recognition from minimalist synthetic binding proteins. AB - Combinatorial libraries built with severely restricted chemical diversity have yielded highly functional synthetic binding proteins. Structural analyses of these minimalist binding sites have revealed the dominant role of large tyrosine residues for mediating molecular contacts and of small serine/glycine residues for providing space and flexibility. The concept of using limited residue types to construct optimized binding proteins mirrors findings in the field of small molecule drug development, where it has been proposed that most drugs are built from a limited set of side chains presented by diverse frameworks. The physicochemical properties of tyrosine make it the amino acid that is most effective for mediating molecular recognition, and protein engineers have taken advantage of these characteristics to build tyrosine-rich protein binding sites that outperform natural proteins in terms of affinity and specificity. Knowledge from preceding studies can be used to improve current designs, and thus synthetic protein libraries will continue to evolve and improve. In the near future, it seems likely that synthetic binding proteins will supersede natural antibodies for most purposes, and moreover, synthetic proteins will enable many new applications beyond the scope of natural proteins. PMID- 19298051 TI - Aminolysis of X-substituted phenyl diphenylphosphinates: effect of amine nature on reactivity and transition-state structure. AB - A kinetic study is reported for aminolysis of X-substituted phenyl diphenylphosphinates (1a-i) in 80 mol % H(2)O/20 mol % dimethyl sulfoxide at 25.0 +/- 0.1 degrees C. The Bronsted-type plot for the reactions of 2,4-dinitrophenyl diphenylphosphinate (1a) with primary amines is linear with beta(nuc) = 0.53. The reactions of 1a-i with ethylamine also result in a linear Bronsted-type plot with beta(lg) = -0.81. These beta(nuc) and beta(lg) values are slightly larger than those reported previously for the reactions of 1a with secondary amines (beta(nuc) = 0.38) and for those of 1a-i with piperidine (beta(lg) = -0.66) but typical for reactions that proceed through a concerted mechanism. It has been concluded that aminolysis of 1a-i proceed through a concerted mechanism and the nature of amines does not affect the reaction mechanism. However, the reactions with primary amines have been suggested to proceed through a later transition state (i.e., more bond formation and bond rupture in the transition state) on the basis of the larger beta(nuc) and beta(lg) values. The concerted mechanism has been further supported from the fact that the Yukawa-Tsuno plot for the reactions of 1a-i with ethylamine exhibits an excellent linear correlation with rho = 2.24 and r = 0.22. Weakly basic primary amines are less reactive than secondary amines of similar basicity. However, strongly basic ethylamine is ca. 2-fold more reactive than piperidine toward 1a, although the former is 0.35 pK(a) units less basic than the latter. PMID- 19298052 TI - Stereodivergent access to polyhydroxylated 10-azabicyclo[4.3.1]decanes as new calystegine analogues. AB - A rapid and stereodivergent access to polyhydroxylated 10 azabicyclo[4.3.1]decanes as new calystegine analogues by way of a double benzotriazolyl/carbon nucleophile exchange followed by a ring-closing metathesis was achieved. Preliminary evaluation of the new compounds as glucocerebrosidase inhibitors was also performed. PMID- 19298053 TI - Quantifying residual stress in nanoscale thin polymer films via surface wrinkling. AB - Residual stress, a pervasive consequence of solid materials processing, is stress that remains in a material after external forces have been removed. In polymeric materials, residual stress results from processes, such as film formation, that force and then trap polymer chains into nonequilibrium stressed conformations. In solvent-cast films, which are central to a wide range of technologies, residual stress can cause detrimental effects, including microscopic defect formation and macroscopic dimensional changes. Since residual stress is difficult to measure accurately, particularly in nanoscale thin polymer films, it remains a challenge to understand and control. We present here a quantitative method of assessing residual stress in polymer thin films by monitoring the onset of strain-induced wrinkling instabilities. Using this approach, we show that thin (>100 nm) polystyrene films prepared via spin-coating possess residual stresses of approximately 30 MPa, close to the crazing and yield stress. In contrast to conventional stress measurement techniques such as wafer curvature, our technique has the resolution to measure residual stress in films as thin as 25 nm. Furthermore, we measure the dissipation of residual stress through two relaxation mechanisms: thermal annealing and plasticizer addition. In quantifying the amount of residual stress in these films, we find that the residual stress gradually decreases with increasing annealing time and plasticizer amounts. Our robust and simple route to measure residual stress adds a key component to the understanding of polymer thin film behavior and will enable identification of more effective processing routes that mitigate the detrimental effects of residual stress. PMID- 19298054 TI - Tracking the relative in vivo pharmacokinetics of nanoparticles with PARACEST MRI. AB - A noninvasive assay that tracks the relative in vivo pharmacokinetics of two nanoparticles may accelerate the development of nanoparticles for biomedical applications, and may provide a method to select personalized nanomedicines for individual patients. To develop an in vivo competitive assay, two MRI contrast agents that could be selectively detected through paramagnetic chemical exchange saturation transfer (PARACEST) were conjugated to a second generation and fifth generation polyamidoamine (PAMAM) dendrimer. The CEST effects of each agent was calibrated relative to concentration. The effects of T(1) relaxivities of these dendritic PARACEST magnetic resonance imaging (MRI) contrast agents were found to be negligible relative to their CEST effects with respect to changes in image contrast, which facilitated the measurement of the ratios of their chemical exchange lifetimes. Injection of both contrast agents into a mouse model of mammary carcinoma resulted in a temporal increase in the CEST effect from each agent in the flank tumor. Although the in vivo CEST effects could not be used to determine the absolute concentrations of each agent within the tumor, the ratio of the in vivo CEST effects was used to measure the ratio of the concentrations of the agents. This result demonstrated that the relative in vivo pharmacokinetics of two nanoparticles may be evaluated using PARACEST MRI. PMID- 19298055 TI - Correcting for electrocatalyst desorption and inactivation in chronoamperometry experiments. AB - Chronoamperometric experiments with adsorbed electrocatalysts are commonly performed either for analytical purposes or for studying the catalytic mechanism of a redox enzyme. In the context of amperometric sensors, the current may be recorded as a function of time while the analyte concentration is being increased to determine a linearity range. In mechanistic studies of redox enzymes, chronoamperometry proved powerful for untangling the effects of electrode potential and time, which are convoluted in cyclic voltammetric measurements, and for studying the energetics and kinetics of inhibition. In all such experiments, the fact that the catalyst's coverage and/or activity decreases over time distorts the data. This may hide meaningful features, introduce systematic errors, and limit the accuracy of the measurements. We propose a general and surprisingly simple method for correcting for electrocatalyst desorption and inactivation, which greatly increases the precision of chronoamperometric experiments. Rather than subtracting a baseline, this consists in dividing the current, either by a synthetic signal that is proportional to the instant electroactive coverage or by the signal recorded in a control experiment. In the latter, the change in current may result from film loss only or from film loss plus catalyst inactivation. We describe the different strategies for obtaining the control signal by analyzing various data recorded with adsorbed redox enzymes: nitrate reductase, NiFe hydrogenase, and FeFe hydrogenase. In each case we discuss the trustfulness and the benefit of the correction. This method also applies to experiments where electron transfer is mediated, rather than direct, providing the current is proportional to the time-dependent concentration of catalyst. PMID- 19298056 TI - DNA-enrichment microfluidic chip for chromatin immunoprecipitation. AB - Chromatin immunoprecipitation (ChIP) is a powerful and widely applied technique for detecting association of individual proteins with specific genomic regions; the technique requires several complex steps and is tedious. In this paper, we develop a microbead-packed microfluidic chip which eliminates most of the laborious, time-consuming, and skill-dependent processes of the ChIP procedure. A computational fluid dynamics model was established to analyze fluidic behavior in a microbead-packed microchannel. With the use of the new chip, a ChIP procedure was performed to purify the GAPDH (glyceraldehyde 3-phosphate dehydrogenase) gene from human embryonic kidney cells (cell line 293). The ChIP capability of the microfluidic chip was evaluated and compared with that of a commercial assay kit; the precipitation performance of both methods was almost identical as shown by quantitative measurement of DNA. However, our chip offers the advantage of low resin volume, and the experimental time is greatly reduced. In addition, our method is less dependent on individual technical skill. Therefore, we expect that our microfluidic chip-based ChIP method will be widely used in DNA-, gene-, and protein-related research and will improve experimental efficiency. PMID- 19298057 TI - Determination of zeta-potential and tortuosity in rat organotypic hippocampal cultures from electroosmotic velocity measurements under feedback control. AB - Extracellular translational motion in the brain is generally considered to be governed by diffusion and tortuosity. However, the brain as a whole has a significant zeta-potential, thus translational motion is also governed by electrokinetic effects under a naturally occurring or applied electric field. We have previously measured zeta-potential and tortuosity in intact brain tissue; however, the method was tedious. In this work, we use a four-electrode potentiostat to control the potential difference between two microreference electrodes in the tissue, creating a constant electric field. Additionally, some alterations have been made to simplify our previous procedure. The method entails simultaneously injecting two 70 kDa dextran conjugated fluorophores into rat organotypic hippocampal cultures and observing their mobility using fluorescence microscopy. We further present two methods of data analysis: regression and two probe analysis. Statistical comparisons are made between the previous and current methods as well as between the two data analysis methods. In comparison to the previous method, the current, simpler method with data analysis by regression gives statistically indistinguishable mean values of zeta-potential and tortuosity, with a similar variability for zeta-potential, -21.3 +/- 2.8 mV, and a larger variability for the tortuosity, 1.98 +/- 0.12. On the other hand, we find that the current method combined with the two-probe analysis produces accurate and more precise results, with a zeta-potential of -22.8 +/- 0.8 mV and a tortuosity of 2.24 +/- 0.10. PMID- 19298058 TI - Single quantum-dot-based aptameric nanosensor for cocaine. AB - Recent advances in single-molecule detection, nanotechnology, and aptameric sensors hold exciting promise for many potential applications. By functionalizing the surface of a quantum dot (QD) with aptamers which can recognize cocaine, and taking advantage of single-molecule detection and fluorescence resonance energy transfer (FRET) between 605QD and Cy5 and Iowa Black RQ, we develop a single-QD based aptameric sensor that is capable of sensing the presence of cocaine through both signal-off and signal-on modes. In comparison with the established aptameric sensors, this single-QD-based aptameric sensor has the significant advantages of simple sample preparation, high sensitivity, and extremely low sample consumption. With the advances in the development of varieties of aptamers for small molecules, nucleic acids, metal ions, and proteins, this single-QD-based aptameric sensor might find wide application in forensic analysis, environmental monitoring, and clinic diagnostics. PMID- 19298059 TI - 1H NMR, GC-EI-TOFMS, and data set correlation for fruit metabolomics: application to spatial metabolite analysis in melon. AB - A metabolomics approach combining (1)H NMR and gas chromatography-electrospray ionization time-of-flight mass spectrometry (GC-EI-TOFMS) profiling was employed to characterize melon (Cucumis melo L.) fruit. In a first step, quantitative (1)H NMR of polar extracts and principal component analyses (PCA) of the corresponding data highlighted the major metabolites in fruit flesh, including sugars, organic acids, and amino acids. In a second step, the spatial localization of metabolites was investigated using both analytical techniques. Direct (1)H NMR profiling of juice or GC-EI-TOFMS profiling of tissue extracts collected from different locations in the fruit flesh provided information on advantages and drawbacks of each technique for the analysis of a sugar-rich matrix such as fruit. (1)H NMR and GC-EI-TOFMS data sets were compared using independently performed PCA and multiblock hierarchical PCA (HPCA), respectively. In addition a correlation-based multiblock HPCA was used for direct comparison of both analytical data sets. These data analyses revealed several gradients of metabolites in fruit flesh which can be related with differences in metabolism and indicated the suitability of multiblock HPCA for correlation of data from two (or potentially more) metabolomics platforms. PMID- 19298060 TI - Electromigration current rectification in a cylindrical nanopore due to asymmetric concentration polarization. AB - We present experimental measurements of electromigration current through a single cylindrical nanopore. A single cylindrical nanopore with 175 nm diameter was fabricated in silicon in series with two micropores with 2 and 100 microm diameters. Thick electrical double layers (EDLs) (kappa a approximately 1) exhibit current rectification due to asymmetric concentration polarization while thinner EDLs show nearly symmetric conductance. After the electric field is turned off, electrical current is measured and observed due to redistribution of ions in the concentration polarization layer. PMID- 19298061 TI - Quantitative CT imaging of the spatial and temporal distribution of liposomes in a rabbit tumor model. AB - Successful employment of noninvasive imaging techniques to quantitatively assess the in vivo pharmacokinetics and biodistribution of nanoparticle drug delivery systems will facilitate the rational design of novel targeted drug carriers. This study reports on the bulk organ/tissue (liver, kidneys, spleen, tumor and blood) and intratumoral distribution of liposomes containing iohexol and gadoteridol over a 14-day period in VX2 sarcoma-bearing New Zealand White rabbits using computed tomography (CT). The vascular half-life of the liposomes was found to be 63.6 +/- 5.8 h and the maximum tumor-to-muscle iodine concentration ratio of 11.9 +/- 6.0 was measured 7 days postinjection with 1.13 +/- 0.29% ID of liposomes accumulating at the tumor site. The liposomes achieved their highest intratumoral distribution volume ratio at 48 h postadministration, occupying 72 +/- 5% of the total tumor volume. This investigation demonstrated the feasibility of using CT to perform quantitative, volumetric and longitudinal assessment of the pharmacokinetics and biodistribution of iodinated liposomes with sensitivities in the range of microg/cm3 while maintaining the ability to identify boundaries of anatomical structures at submillimeter resolution and with imaging time of less than one minute per scan. If successfully approved for clinical adoption, the use of CT imaging to monitor nanoparticulate drug delivery will provide an opportunity for online adjustment of therapeutic regimens and implementation of personalized medicine. PMID- 19298062 TI - Real-time NMR investigations of structural changes in silicon electrodes for lithium-ion batteries. AB - Lithium-ion batteries (LIBs) containing silicon negative electrodes have been the subject of much recent investigation because of the extremely large gravimetric and volumetric capacity of silicon. The crystalline-to-amorphous phase transition that occurs on electrochemical Li insertion into crystalline Si, during the first discharge, hinders attempts to link structure in these systems with electrochemical performance. We apply a combination of static, in situ and magic angle sample spinning, ex situ (7)Li nuclear magnetic resonance (NMR) studies to investigate the changes in local structure that occur in an actual working LIB. The first discharge occurs via the formation of isolated Si atoms and smaller Si Si clusters embedded in a Li matrix; the latter are broken apart at the end of the discharge, forming isolated Si atoms. A spontaneous reaction of the lithium silicide with the electrolyte is directly observed in the in situ NMR experiments; this mechanism results in self-discharge and potential capacity loss. The rate of this self-discharge process is much slower when CMC (carboxymethylcellulose) is used as the binder. PMID- 19298063 TI - Using biological performance similarity to inform disaccharide library design. AB - Designing better small-molecule discovery libraries requires having methods to assess the consequences of different synthesis decisions on the biological performance of resulting library members. Since we are particularly interested in how stereochemistry affects performance in biological assays, we prepared a disaccharide library containing systematic stereochemical variations, assayed the library for different biological effects, and developed methods to assess the similarity of performance between members across multiple assays. These methods allow us to ask which subsets of stereochemical features best predict similarity in patterns of biological performance between individual members and which features produce the greatest variation of outcomes. We anticipate that the data analysis approach presented here can be generalized to other sets of biological assays and other chemical descriptors. Methods to assess which structural features of library members produce the greatest similarity in performance for a given set of biological assays should help prioritize synthesis decisions in second-generation library development targeting the underlying cell-biological processes. Methods to assess which structural features of library members produce the greatest variation in performance should help guide decisions about what synthetic methods need to be developed to make optimal small-molecule screening collections. PMID- 19298064 TI - Isolation and characterization of an aggregating peptide from a tryptic hydrolysate of whey proteins. AB - Spontaneous precipitation of a peptide mixture has been observed during the concentration by reverse osmosis of a tryptic hydrolysate of whey protein. The precipitated material collected by centrifugation could not be solubilized by urea, mercaptoethanol, or sodium dodecyl sulfate. However, a complete solubilization of the aggregates was observed when the pH of the solution was lowered to 2.0. Analysis of the insoluble fraction has allowed the identification of beta-lactoglobulin (beta-lg) fragment 1-8 as the major peptide involved in the formation of aggregates. Peptide beta-lg f1-8 accounted for >94% of the peptide content in the precipitate washed twice with distilled water. The investigation of the secondary structure using circular dichroism evidenced that the peptide beta-lg f1-8 isolated from the flocculated peptide mixture is under random coil conformation at acidic and neutral pH and tends to adopt a beta-sheet conformation at basic pH. The findings of this study provide evidence that peptide beta-lg f1-8 forms aggregates via an efficient self-assembly process. PMID- 19298065 TI - Botanical and geographical characterization of green coffee (Coffea arabica and Coffea canephora): chemometric evaluation of phenolic and methylxanthine contents. AB - Green coffee beans of the two main commercial coffee varieties, Coffea arabica (Arabica) and Coffea canephora (Robusta), from the major growing regions of America, Africa, Asia, and Oceania were studied. The contents of chlorogenic acids, cinnamoyl amides, cinnamoyl glycosides, free phenolic acids, and methylxanthines of green coffee beans were analyzed by liquid chromatography coupled with UV spectrophotometry to determine their botanical and geographical origins. The analysis of caffeic acid, 3-feruloylquinic acid, 5-feruloylquinic acid, 4-feruloylquinic acid, 3,4-dicaffeoylquinic acid, 3-caffeoyl-5 feruloylquinic acid, 3-caffeoyl-4-feruloylquinic acid, 3-p-coumaroyl-4 caffeoylquinic acid, 3-caffeoyl-4-dimethoxycinnamoylquinic acid, 3-caffeoyl-5 dimethoxycinnamoylquinic acid, p-coumaroyl-N-tryptophan, feruloyl-N-tryptophan, caffeoyl-N-tryptophan, and caffeine enabled the unequivocal botanical characterization of green coffee beans. Moreover, some free phenolic acids and cinnamate conjugates of green coffee beans showed great potential as means for the geographical characterization of coffee. Thus, p-coumaroyl-N-tyrosine, caffeoyl-N-phenylalanine, caffeoyl-N-tyrosine, 3-dimethoxycinnamoyl-5 feruloylquinic acid, and dimethoxycinnamic acid were found to be characteristic markers for Ugandan Robusta green coffee beans. Multivariate data analysis of the phenolic and methylxanthine profiles provided preliminary results that allowed showing their potential for the determination of the geographical origin of green coffees. Linear discriminant analysis (LDA) and partial least-squares discriminant analysis (PLS-DA) provided classification models that correctly identified all authentic Robusta green coffee beans from Cameroon and Vietnam and 94% of those from Indonesia. Moreover, PLS-DA afforded independent models for Robusta samples from these three countries with sensitivities and specificities of classifications close to 100% and for Arabica samples from America and Africa with sensitivities of 86 and 70% and specificities to the other class of 90 and 97%, respectively. PMID- 19298066 TI - Spectroscopic and computational characterization of the base-off forms of cob(II)alamin. AB - The one-electron-reduced form of vitamin B(12), cob(II)alamin (Co(2+)Cbl), is found in several essential human enzymes, including the cobalamin-dependent methionine synthase (MetH). In this work, experimentally validated electronic structure descriptions for two "base-off" Co(2+)Cbl species have been generated using a combined spectroscopic and computational approach, so as to obtain definitive clues as to how these and related enzymes catalyze the thermodynamically challenging reduction of Co(2+)Cbl to cob(I)alamin (Co(1+)Cbl). Specifically, electron paramagnetic resonance (EPR), electronic absorption (Abs), and magnetic circular dichroism (MCD) spectroscopic techniques have been employed as complementary tools to characterize the two distinct forms of base-off Co(2+)Cbl that can be trapped in the H759G variant of MetH, one containing a five coordinate and the other containing a four-coordinate, square-planar Co(2+) center. Accurate spin Hamiltonian parameters for these low-spin Co(2+) centers have been determined by collecting EPR data using both X- and Q-band microwave frequencies, and Abs and MCD spectroscopic techniques have been employed to probe the corrin-centered pi --> pi* and Co-based d --> d excitations, respectively. By using these spectroscopic data to evaluate electronic structure calculations, we found that density functional theory provides a reasonable electronic structure description for the five-coordinate form of base-off Co(2+)Cbl. However, it was necessary to resort to a multireference ab initio treatment to generate a more realistic description of the electronic structure of the four-coordinate form. Consistent with this finding, our computational data indicate that, in the five coordinate Co(2+)Cbl species, the unpaired spin density is primarily localized in the Co 3d(z(2))-based molecular orbital, as expected, whereas in the four coordinate form, extensive Co 3d orbital mixing, configuration interaction, and spin-orbit coupling cause the unpaired electron to delocalize over several Co 3d orbitals. These results provide important clues to the mechanism of enzymatic Co(2+)Cbl --> Co(1+)Cbl reduction. PMID- 19298067 TI - Combined experimental studies and theoretical calculations to yield the complete molecular structure and vibrational spectra of (CH3)3GeH. AB - The molecular structure of trimethylgermane has been determined by gas electron diffraction experiments. Infrared spectra for the gaseous, liquid, and solid phases were also recorded. Parallel and perpendicular polarized Raman spectra for the liquid were measured to obtain depolarization values. The experimental studies were supported by a series of computational calculations using HF, B3LYP, and MP2 methods and a variety of basis sets. The force fields obtained from density functional theory using both B3LYP/6-31G* and B3LYP/6-311+G** were scaled with both Pulay's SQM methodology and Yoshida's WLS procedure to simulate the vibrational spectra and assist in the assignment of fundamental bands. The Raman intensities were obtained from polarizability derivatives. The vibrational spectra of trimethylgermane were completely assigned on the basis of the experimental data and the theoretical prediction of vibrational frequencies and intensities. PMID- 19298068 TI - Quantitative structure-property relationship estimation of cation binding affinity of the common amino acids. AB - The quantitative structure-property relationship (QSPR) methodology is applied to estimate the binding affinity of lithium, sodium, potassium, copper, and silver cations to the 20 common amino acids. The proposed model, nonlinearly derived from computational neural networks (CNN), contains seven descriptors and was validated by an external prediction set. Good results are obtained with correlation coefficients, R(2), and root-mean-square errors (rms) (kJ/mol) of 0.998 (3.89), 0.999 (2.86), and 0.997 (3.90) for the training, prediction, and validation sets, respectively. Five of the descriptors of the model correspond to the amino acids and the other two to the cations; they encode information clearly related to the cation-amino acid interactions responsible for the binding affinity values analyzed. A detailed analysis of results shows that, despite the different nature of the bonding between the metal cations and the amino acids, the neural networks used are capable of predicting accurately the property studied. PMID- 19298069 TI - Hygroscopic growth and deliquescence of NaCl nanoparticles coated with surfactant AOT. AB - Aerosolized nanoparticles of NaCl coated with variable amounts of surfactant AOT were generated by electrospraying AOT/NaCl aqueous solutions, followed by neutralizing and drying the resulting particles. A tandem differential mobility analyzer was used to select a narrow size distribution of particles with mobility equivalent diameters below 20 nm and monitor their hygroscopic growth as a function of relative humidity. Effects of the particle size and relative amount of surfactant on the hygroscopic growth of NaCl were studied. For pure NaCl nanoparticles, the deliquescence relative humidity (DRH) increased as the particle size was decreased, in full agreement with previous measurements. Below the DRH the NaCl nanoparticles had an equivalent of one-four monolayers of water adsorbed on the surface. The addition of a sub-monolayer AOT coating reduced the DRH and suppressed the hygroscopic growth of the NaCl core. At AOT coverage levels exceeding one monolayer, a clear deliquescence transition was no longer discernible. The Zdanovskii-Stokes-Robinson (ZSR) model failed to predict the observed growth factors of mixed AOT/NaCl nanoparticles reflecting a large contribution of the interfacial interactions between NaCl and AOT to the total free energy of the particles. There were indications that AOT/NaCl nanoparticles prepared by the electrospray aerosol source were enhanced in the relative mass fraction of AOT in comparison with the solution from which they were electrosprayed. PMID- 19298070 TI - Crystal structure of the acid-induced arginine decarboxylase from Escherichia coli: reversible decamer assembly controls enzyme activity. AB - The acid-induced arginine decarboxylase is part of an enzymatic system in Escherichia coli that contributes to making this organism acid resistant. The arginine decarboxylase is a vitamin B(6)-dependent enzyme that is active at acidic pH. It consumes a proton in the decarboxylation of arginine to agmatine, and by working in tandem with an arginine-agmatine antiporter, this enzymatic cycle protects the organism by preventing the accumulation of protons inside the cell. We have determined the structure of the acid-induced arginine decarboxylase by X-ray crystallography to 2.4 A resolution. The arginine decarboxylase structure revealed a ca. 800 kDa decamer composed as a pentamer of five homodimers. Each homodimer has an abundance of acidic surface residues, which at neutral pH prevents inactive homodimers from associating into active decamers. Conversely, acidic conditions favor the assembly of active decamers. Therefore, the structure of arginine decarboxylase presents a mechanism by which its activity is modulated by external pH. PMID- 19298071 TI - Nonadiabatic ab initio dynamics of two models of Schiff base retinal. AB - On-the-fly classical dynamics calculations combined with ab initio quantum chemical computations are carried out for two models of protonated Schiff base retinal in vacuo. The models are the 6pi system of 2-cis-penta-2,4-dieneimminium cation and the 12pi system in which two methyl groups are removed from the Schiff base of retinal. The CASSCF(6,6) level with the 6-31G basis set was employed for the quantum chemical part and the velocity Verlet algorism is utilized for time evolution of trajectories. The probabilities of nonadiabatic transition between the excited and ground state are estimated by the Zhu-Nakamura formulas. The 9 cis form product in addition to the all-trans one is generated in the present gas phase calculation for the 12pi model, despite the 9-cis generation being suppressed in protein. We have found that energy relaxation on the ground state occurs in two steps in the 12pi model. In the first step a metastable intermediate state is formed at approximately 100 fs after photoexcitation at the energy around 20-40 kcal/mol down from the excited potential energy surface, then it further relaxes to the energy around 60-80 kcal/mol from the excited surface, leading to the final state (second step). This relaxation pattern can be seen in all the three pathways to the all-trans, 9-cis, and (reverted) 11-cis form. Fourier transformation analysis reveals that the effective vibrational frequencies of the intermediate state are 1600-2000 cm(-1), which can be attributed to the conjugate CC bond frequencies in the electronic ground state. The two-step relaxation may be due to dynamical barriers. The two-step relaxation is not revealed in the smaller 6pi model. The crank-shaft motion of the C11C12 and C9C10 bonds is found in the isomerization, which indicates the motion is intrinsic in retinal, not due to the surrounding protein. The branching ratio is about 1:1:2 for the all-trans, 9-cis, and 11-cis form generation. The ratio is different from earlier works where Tully's fewest switching scheme was employed. The bond length and the dihedral angle at the transitions are also analyzed to investigate the transition mechanism. PMID- 19298072 TI - Relative tropospheric photolysis rates of acetaldehyde and formaldehyde isotopologues measured at the European photoreactor facility. AB - The photolysis rates of HCHO, DCDO, CH(3)CHO, and CH(3)CDO are studied by long path FTIR spectroscopy in natural tropospheric conditions at the European Photoreactor Facility (EUPHORE) in Valencia, Spain. Average relative photolysis rates j(HCHO)/j(DCDO) = 3.15 +/- 0.08 and j(CH(3)CHO)/j(CH(3)CDO) = 1.26 +/- 0.03 are obtained from three days of experiments for each reaction in the period June 17 to July 7, 2006. PMID- 19298073 TI - Facile and selective procedure for the synthesis of bridged 1,2,4,5-tetraoxanes; strong acids as cosolvents and catalysts for addition of hydrogen peroxide to beta-diketones. AB - A facile, experimentally simple, and selective method was developed for the synthesis of bridged 1,2,4,5-tetraoxanes based on the reaction of hydrogen peroxide with beta-diketones catalyzed by strong acids (H(2)SO(4), HClO(4), HBF(4), or BF(3)). The yields of the target products vary from 44% to 77%. 1,2,4,5-Tetraoxanes can easily be separated from other reaction products by column chromatography. A high concentration of a strong acid is a key factor determining the selectivity of formation and the yield of 1,2,4,5-tetraoxanes. Unlike many compounds containing the O-O bond, which undergo rearrangements in acidic media, the resulting cyclic peroxides are quite stable under these conditions. PMID- 19298074 TI - Prediction of remarkably large second-order nonlinear optical properties of organoimido-substituted hexamolybdates. AB - The dipole polarizabilities, dipole moments, density of states, and second-order nonlinear optical (NLO) properties of organoimido derivatives of hexamolybdates have been investigated by using time-dependent density functional response theory. This class of organic-inorganic hybrid compounds possesses remarkably large and eye-catching molecular second-order NLO response, especially [Mo(6)O(17)(NC(16)H(12)NO(2))(FeNC(10)H(9))](2-) (7) and [Mo(6)O(17)(NC(16)H(12)NO(2))(NC(6)H(2)(NH(2))(3))](2-) (6) with static second order polarizability (beta(vec)) computed to be 15766.27 x 10(-30) esu and 6299.59 x 10(-30) esu, respectively. Thus, these systems have the possibility to be excellent second-order nonlinear optical materials. Analysis of the major contributions to the beta(vec) value suggests that the charge transfer (CT) from polyanion to organic segment (D-A) along the z-axis plays the key role in NLO response; the polyanion acts as a donor (D) whereas organoimido acts as an acceptor (A) in all the studied systems. The computed beta(vec) values increase by incorporation of an electron acceptor (-NO(2)) at the end of the phenyl ring of the organoimido segment. Furthermore, substitution of amino (-NH(2)) or ferrocenyl (-FeC(10)H(9)) at the outer side of polyanion and an electron acceptor (-NO(2)) at the end of the phenyl ring in organoimido segment simultaneously is more important to enhance the optical nonlinearity. Orbital analysis shows that the degree of CT between the polyanion and organoimido segments was increased when ferrocenyl donor was introduced. The present investigation provides important insight into the remarkably large NLO properties of organoimido substituted hexamolybdates. PMID- 19298076 TI - CLiDE Pro: the latest generation of CLiDE, a tool for optical chemical structure recognition. AB - We present CLiDE Pro, the latest version of the output of the long-term CLiDE project for the development of tools for automatic extraction of chemical information from the literature. CLiDE Pro is concerned with the extraction of chemical structure and generic structure information from electronic images of chemical molecules available online as well as pages of scanned chemical documents. The information is extracted in three phases, first the image is segmented into text and graphical regions, then graphical regions are analyzed and where possible the connection tables are reconstructed, and finally any generic structures are interpreted by matching R-groups found in structure diagrams with the ones located in the text. The program has been tested on a large set of images of chemical structures originating from various sources. The results demonstrate good performance in the reconstruction of connection tables with few errors in the interpretation of the individual drawing features found in the structure diagrams. This full test set is presented for use in the validation of other similar systems. PMID- 19298077 TI - Effect of phase structure on enzymatic degradation in poly(L-lactide)/atactic poly(3-hydroxybutyrate) blends with different miscibility. AB - Thin films of poly(L-lactide) (PLLA)/atactic poly(3-hydroxybutyrate) (ataPHB) blends with different miscibility were prepared and characterized by using differential scanning calorimetry (DSC) and atomic force microscopy (AFM). The DSC analysis suggested that the blend thin films exhibited different phase structures, such as miscible, partially miscible, and immiscible depending on the blending ratio as well as molecular weight of ataPHB component. The different miscibility was further confirmed by the surface morphological observation by AFM. Both the immiscible and partially miscible blends of PLLA/ataPHB revealed the formation of phase-separated morphology of PLLA and ataPHB components, whereas the homogeneous surface morphology was observed for the miscible blend. On the basis of the changes in the depth profile from the surface level of the thin films, the enzymatic degradation rates of the PLLA and ataPHB domains were determined in the presence of either PHB depolymerase or proteinase K, respectively. The erosion rate of PLLA/ataPHB blends was strongly dependent on the blend composition and the degree of dispersion of the two components. The enzymatic degradation behaviors were discussed in terms of phase structure, molecular mobility, and retardation effect of the components in the blends. PMID- 19298078 TI - Biodegradable materials based on silk fibroin and keratin. PMID- 19298079 TI - Fumonisin determination in tunisian foods and feeds. ELISA and HPLC methods comparison. AB - A survey for fumonisins B1 and B2 (FB1 and FB2, respectively) was performed on 180 samples of high-consumption food commodities and 15 feed samples randomly collected from various regions of Tunisia. The determination of fumonisin level was performed by an in-house validated high-performance liquid chromatography and an enzyme-linked immunosorbent assay (ELISA) methods. Detection limit by ELISA for fumonisins sum was 25 microg/kg, and those by HPLC were 50 microg/kg for FB1 and 70 microg/kg for FB2. Recoveries of fumonisins spiked at 130 microg/kg ranged from 68.5 to 75.6% by ELISA, whereas those by HPLC for FB1 at 400 microg/kg and for FB2 at 300 microg/kg varied from 75.2 to 90.5%. Naturally occurring fumonisins were found in 10.5% of food samples with levels ranging from 70 to 2130 microg/kg. All contaminated samples contained FB1, and 31.5% of them contained FB2. In addition, the most contaminated commodities were corn foods and sorghum, whereas no fumonisin contamination was found in any nut or rice samples. For analyzed feed samples, fumonisins were detected in 86.6% of them with concentrations ranging from 50 to 2800 microg/kg. In addition, the performance of analytical methods was investigated in a comparison between ELISA and HPLC results for samples analyzed by both methods. PMID- 19298080 TI - Synthesis, crystal and electronic structures, and physical properties of the novel compounds LaR4Mo36O52 (R = Dy, Er, Yb, and Y) containing infinite chains of trans-edge-shared Mo6 octahedra and Mo2 pairs and rectangular Mo4 clusters with triple Mo-Mo bonds. AB - The novel quaternary reduced molybdenum oxides LaR(4)Mo(36)O(52) (R = Dy, Er, Yb, and Y) have been synthesized with solid-state reactions at 1400 degrees C for 48 h in sealed molybdenum crucibles. The crystal structure was determined on a single crystal of LaEr(4)Mo(36)O(52) by X-ray diffraction. LaEr(4)Mo(36)O(52) crystallizes in the tetragonal space group I4 with two formula units per cell and the following lattice parameters: a = 19.8348(2) and c = 5.6594(1) A. The Mo network is dominated by infinite chains of trans-edge-shared Mo(6) octahedra, which coexist with Mo(2) pairs and rectangular Mo(4) clusters. The Mo-Mo distances within the infinite chains range from 2.5967(7) to 2.8529(8) A and from 2.239(3) to 2.667(2) A in the Mo(2) pairs and rectangular Mo(4) clusters, respectively. The Mo-O distances are comprised between 1.993(7) and 2.149(7) A, as usually observed in these types of compound. The La(3+) and Er(3+) ions are in a square-prismatic [LaO(8)] and a tricapped trigonal-prismatic [ErO(9)] environment of oxygen atoms, respectively. The La-O distances range from 2.555(6) to 2.719(6) A and the Er-O ones from 2.260(6) to 2.469(5) A. Theoretical calculations allow the determination of the optimal electron count of both motifs in the title compound. Weak interactions occur between neighboring dimetallic and tetrametallic clusters and between trans-edge-sharing infinite chains and dimers and tetramers. The presence of rectangular clusters is favored on the basis of theoretical considerations. Single-crystal resistivity measurements show that LaEr(4)Mo(36)O(52) is metallic between 4.2 and 300 K, in agreement with the band structure calculations. Magnetic susceptibility measurements indicate that the oxidation state of the magnetic rare earths is +3, and there is an absence of localized moments on the Mo network. PMID- 19298081 TI - Efficient fragment coupling approaches toward large oxacalix[n]arenes (n = 6, 8). AB - The first rational, stepwise synthesis of enlarged oxacalix[n]arenes (n > 4) is described. Variously substituted oxacalix[3]arene[3]pyrimidines were prepared rather selectively by a straightforward [3 + 3] fragment coupling approach after a thorough search for the optimum nucleophilic aromatic substitution conditions. Similar procedures also allowed facile synthesis of unsymmetrical oxacalix[4]- and oxacalix[8]arenes. PMID- 19298082 TI - Visualization of high-dimensional combinatorial catalysis data. AB - The role of various techniques for visualization of high-dimensional data is demonstrated in the context of combinatorial high-throughput experimentation (HTE). Applying visualization tools, we identify which constituents of catalysts are associated with final products in a huge combinatorially generated data set of heterogeneous catalysts, and catalytic activity regions are identified with respect to pentanary composition spreads of catalysts. A radial visualization scheme directly visualizes pentanary composition spreads in two-dimensional (2D) space and catalytic activity of a final product by combining high-throughput results from five slate libraries. A glyph plot provides many possibilities for visualizing high-dimensional data with interactive tools. For catalyst discovery and lead optimization, this work demonstrates how large multidimensional catalysis data sets are visualized in terms of quantitative composition activity relationships (QCAR) to effectively identify the relevant key role of compositions (i.e., lead compositions) of catalysts. PMID- 19298083 TI - Feshbach resonances: the branching of quantum mechanics into Hermitian and non Hermitian formalisms. AB - As has been shown long time ago by Feshbach, the exact energy spectrum of the full problem can be obtained by solving two different self-energy problems. In spite of the fact that the two effective Hamiltonians are derived in very similar ways in one case, the exact energy spectrum of the full problem can be either real or complex (depending on the boundary conditions), whereas the exact energy spectrum associated with the second effective Hamiltonian has to be complex (excluding bound states in the continuum). The focus of this paper is on the fact that in both cases the complex eigenvalues result from the same requirement of an out-going boundary condition. The branching of quantum mechanics to standard (Hermitian) formalism and non-Hermitian formalism is associated with the decision to express the exact energy spectrum with one of the two possible self-consistent like problems where the use of the Green operator imposes an outgoing boundary condition on the solutions of the time-independent Schrodinger equation. Our analysis is made for the case where an ABC molecule has sufficient energy to dissociate to A + BC but not to A + B + C and not to AB + C or to AC + B. PMID- 19298084 TI - Capillary-based instrument for the simultaneous measurement of solution viscosity and solute diffusion coefficient at pressures up to 2000 bar and implications for ultrahigh pressure liquid chromatography. AB - An instrument based on the Poiseuille flow principle capable of measuring solution viscosities at high pressures has been modified to observe UV-absorbent analytes in order to allow for the simultaneous measurement of analyte diffusivity. A capillary time-of-flight (CTOF) instrument was used to measure the viscosity of acetonitrile-water mixtures in all decade volume percent increments and the corresponding diffusion coefficients of small aromatic molecules in these solvent mixtures from atmospheric pressure to 2000 bar (approximately 30,000 psi) at 25 degrees C. The instrument works by utilizing a relatively small pressure drop (<100 bar) across a fused-silica capillary which has both the inlet and outlet pressurized so that the average column pressure can be significantly elevated (up to 2000 bar). Measurements with this instrument agree with high pressure viscosity data collected previously using a CTOF viscometer, as well as with literature values obtained with falling-body viscometers of the Bridgman design. It has been further determined that, for the small molecules included in this study, trends in solute diffusivity with respect to pressure follow the predictions of the Stokes-Einstein equation when the solvent viscosity is corrected as a function of pressure. Because the instrument described herein determines viscosity and diffusivity independently, the effect of pressure on analyte hydrodynamic radius can also be monitored. An analysis of ultrahigh pressure liquid chromatography (UHPLC) data was performed using the pressure corrected diffusion coefficient of hydroquinone to demonstrate the effect of this phenomenon on the analysis of chromatographic performance. PMID- 19298085 TI - Interrogating the molecular details of the peroxiredoxin activity of the Escherichia coli bacterioferritin comigratory protein using high-resolution mass spectrometry. AB - Bacterioferritin comigratory protein (BCP) is a bacterial thioredoxin-dependent thiol peroxidase that reduces a variety of peroxide substrates. Using high resolution Fourier transform ion cyclotron resonance mass spectrometry coupled with top-down fragmentation techniques, we have analyzed the mechanistic details of hydrogen peroxide reduction by E. coli BCP. We show here that catalysis occurs via an atypical two-cysteine peroxiredoxin pathway. A transient sulfenic acid is initially formed on Cys-45, before resolution by the formation of an intramolecular disulfide bond between Cys-45 and Cys-50. This oxidized BCP intermediate is shown to be a substrate for reduction by thioredoxin, completing the catalytic cycle. Although we invoke Cys-50 in the catalytic cycle of Escherichia coli bacterioferritin comigratory protein (BCP), a previous study had shown that this residue was not absolutely required for peroxiredoxin activity. In order to explain these apparently conflicting phenomena, we analyzed the reaction of a C50S BCP mutant with peroxide. We show that this mutant BCP enzyme adopts a different and novel mechanistic pathway. The C50S BCP mutant reacts with peroxide to form a sulfenic acid on Cys-45, in the same manner as wild-type BCP. However, the nascent intermediate is then resolved by reaction with Cys-45 from a second BCP molecule, resulting in a dimeric intermediate containing an intermolecular disulfide bond. We further show that this novel resolving complex is a substrate for reduction by thioredoxin. The importance of our results in furthering the understanding of catalysis within BCP family is discussed. PMID- 19298087 TI - Regenerating science: stem cells back in the mix. PMID- 19298091 TI - Sheng Ding: using chemical methods to control cell fate. AB - Profiles provide insights into the lives, backgrounds, career paths, and futures of scientists who serve as Experts on ACS Chemical Biology's online Ask the Expert feature. Ding will begin answering your questions in mid-March, 2009. Readers are encouraged to submit their questions to the Experts at http://community.acs.org/chembiol/ . The editors will post the most interesting exchanges on the web site. PMID- 19298086 TI - Illuminating the chemistry of life: design, synthesis, and applications of "caged" and related photoresponsive compounds. AB - Biological systems are characterized by a level of spatial and temporal organization that often lies beyond the grasp of present day methods. Light modulated bioreagents, including analogs of low molecular weight compounds, peptides, proteins, and nucleic acids, represent a compelling strategy to probe, perturb, or sample biological phenomena with the requisite control to address many of these organizational complexities. Although this technology has created considerable excitement in the chemical community, its application to biological questions has been relatively limited. We describe the challenges associated with the design, synthesis, and use of light-responsive bioreagents; the scope and limitations associated with the instrumentation required for their application; and recent chemical and biological advances in this field. PMID- 19298092 TI - Expanding the repertoire of fluorescent calcium sensors. AB - Fluorescent indicators for calcium are incredibly powerful because they enable researchers to watch the movement of calcium ions in real time in living cells. The popular small molecule indicator Indo-1 has now been targeted to a defined location, namely, the nucleus of muscle cells, using SNAP-tag technology. This combination of a chemical probe with genetic targeting expands the available options for measuring local calcium events in cells. PMID- 19298093 TI - Physical aging and phase behavior of multiresponsive microgel colloidal dispersions. AB - Quantitative microscopy measurements have been made on poly(N-isopropylacrylamide co-acrylic acid) (pNIPAm-AAc) microgel dispersions as a function of time, temperature, pH, and volume fraction. These studies reveal an extreme degree of complexity in the physical aging and phase behavior of the dispersions; this complexity arises from a convolution of the system energetics at the colloidal, polymer-chain, and molecular scales. Superficially, these dispersions display the classic colloidal phases observed for spherical particles (i.e., gas, fluid, crystal, and glass). However, unlike simple repulsive hard spheres, pNIPAm-AAc dispersions are observed to evolve from a diffusive, fluidlike state immediately after being introduced into rectangular capillary tubes, to very slow crystalline or glassy phases after days or weeks of aging. In addition to this structural evolution, the free volume accessible to the microgels in crystalline or glassy phases (i.e., the cage size) decreases with time, indicating that the physical aging process does not end following assembly, but instead continues to evolve as the dispersion slowly proceeds to an equilibrium state. The temperature dependence of pNIPAm-AAc microgel swelling and how it influences the colloidal assembly was evaluated during the aging process as well. These thermal melting experiments revealed an enhancement in the thermal stability (i.e., a decrease in the influence of temperature on the phase behavior) of the assemblies during the aging process that we associate with an evolution of attractive interparticle interactions during aging. These attractive interactions dictate the time scale for assembly (aging), the final phase adopted by the dispersion, the dynamics of the final state, and the ultimate thermal stability. The culmination of these studies is the pseudoequilibrium phase behavior of pNIPAm-AAc microgel dispersions, which we present as a function of pH and volume fraction following approximately 1 month of aging. This diagram reveals highly complex dispersion characteristics that appear to be intrinsically tied to the degree of AAc protonation. In general, we find that, at pH < pK(a), the final dispersions behave in a manner that can be associated with attractive interparticle interactions, whereas at pH > pK(a), repulsive interactions appear to be dominant. These results are discussed in the context of the slow evolution of microgel swelling and attractive interaction potentials arising from reorganization and association of polymer chains via multiple weak hydrogen bonding interactions. PMID- 19298094 TI - Patient perspectives on primary health care in rural communities: effects of geography on access, continuity and efficiency. AB - INTRODUCTION: Examining how to deliver primary health care (PHC) services and increase their accessibility (regardless of geographic location) from the patient's perspective is needed. We conducted seven focus groups with people (n = 50) living in rural communities, in British Columbia, Canada, as they reflected on priorities for and use of PHC. METHODS: In addition to discussing their priorities for PHC services, participants completed a brief questionnaire designed to collect information regarding socio-demographics, health status and utilization of primary healthcare providers. Descriptive statistics were obtained from questionnaire data. Focus group data were coded using an evaluation framework specifically developed for PHC; a thematic content analysis was then conducted on the coded data. RESULTS: In total, 80% of participants had been patients of the same provider for more than one year and had an average of two chronic conditions. Participants described the challenges posed by geographical location in terms of: (1) making tradeoffs; (2) management, information, and relationship continuity of care; and (3) efficiency with health care delivery. Additional out-of-pocket expenses were associated with traveling to regional centers for health services. Those living in rural communities, especially people needing additional health services to manage their health problems, made tradeoffs between their safety of having to travel during times of poor road conditions and having their healthcare needs met. CONCLUSION: Challenges to timely access to a regular healthcare provider, continuity of information and management of people's chronic disease conditions, and linkages to specialist services and diagnostic tests pose challenges for those living in rural communities. The geographic location of rural communities compounds the extent to which these people are able to access timely and continuous PHC. PMID- 19298095 TI - VHA pharmacy use in veterans with Medicare drug coverage. AB - OBJECTIVE: To determine how Medicare benefits affect veterans' use of Veterans Health Administration (VHA) pharmacy services. STUDY DESIGN: Retrospective analysis of veterans dually enrolled in the Veterans Health Administration and Medicare healthcare systems. METHODS: We used VHA and Medicare administrative data for calendar year 2002 to examine the effect of Medicare HMO pharmacy benefit levels on VHA pharmacy use. RESULTS: In 2002, 64% of the VHA and Medicare dually enrolled veterans in our study sample received medications from the VHA. Use of VHA pharmacy services varied monotonically by the level of pharmacy benefits among Medicare HMO enrollees, with veterans enrolled in plans with both low and high pharmacy benefit levels significantly less likely to use VHA pharmacy services than veterans in plans with no pharmacy benefits (odds ratios = .83 and .53, respectively, versus plans with no benefits). Among VHA pharmacy users, enrollment in plans with high levels of benefits was associated with significantly lower annual pharmacy costs than enrollment in plans with no benefits or enrollment in traditional Medicare. CONCLUSIONS: Our findings indicate that non-VHA pharmacy benefits affect both the likelihood and magnitude of VHA pharmacy use. This suggests that Medicare pharmacy coverage (Part D) may significantly reduce the demand for VHA pharmacy services, particularly in geographic regions previously underserved by Medicare managed care plans. PMID- 19298096 TI - Voice response system to measure healthcare costs: a STAR*D report. AB - OBJECTIVE: To evaluate a telephone-operated, interactive voice response (IVR) system designed to collect use-of-care data from patients with major depression (UAC-IVR). STUDY DESIGN: Patient self-reports from repeated IVR surveys were compared with provider records for 3789 patients with major depression at 41 clinical sites participating in the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) trial. METHODS: UAC-IVR responses were examined for consistency and compared with provider records to compute reporting biases and intraclass correlation coefficients. Predictors of inconsistent responses and reporting biases were based on mixed logistic and regression models adjusted for need and predisposing and enabling covariates, and corrected for nesting and repeated measures. RESULTS: Inconsistent responses were found for 10% of calls and 21% of patients. Underreporting biases (-20%) and moderate agreement (intraclass correlation of 68%) were found when UAC-IVR responses were compared with medical records. IVR reporting biases were less for patients after 3 calls or more (experience), for patients with severe baseline symptoms (motivation), and for patients who gave consistent IVR responses (reliability). Bias was unrelated to treatment outcomes or demographic factors. CONCLUSION: Clinical managers should use IVR systems to collect service histories only after patients are properly trained and responses monitored for consistency and reporting biases. PMID- 19298097 TI - The Vermedx Diabetes Information System reduces healthcare utilization. AB - OBJECTIVE: To confirm the cost savings in a randomized clinical trial of the Vermedx Diabetes Information System (hereafter referred to as the Diabetes Information System [DIS]) in independently collected data using claims paid by a managed care insurer for patients with and without DIS participation. STUDY DESIGN: Longitudinal analysis of paid claims with concurrent and historical controls from October 2002 through October 2007. METHODS: Using locally weighted smoothing functions and linear regression analysis before and after commencement of the DIS, we compared the total claims paid per member per month for 153 patients using the DIS versus 870 control patients. RESULTS: For DIS patients, paid claims increased at a rate of $8.30 (95% confidence interval [CI], $1.12 $15.48) per month before the DIS started compared with -$3.92 (95% CI, -$9.50 to $1.67) after commencement of the DIS (P = .008). For control patients, the slope changed from $6.80 (95% CI, $3.78-$9.82) to $3.16 (95% CI, -$1.06 to $7.38) (P = .17). After commencement of the DIS, the slope of the claims in the DIS group is significantly lower than that of the control group (-$3.92 vs $3.16, P = .046). The mean estimated savings range from $504 per patient in year 1 of operations to $3563 in year 4. The cumulative net savings reach $8134 in 4 years. CONCLUSIONS: Participation in the DIS is associated with substantial reductions in claims paid, net of the costs of the intervention. The cost savings reported in the randomized clinical trial of the DIS are reproduced in an independent data set. PMID- 19298098 TI - Curtailing laboratory test ordering in a managed care setting through redesign of a computerized order form. AB - OBJECTIVE: To increase appropriate use of blood tests for folic acid and vitamin B12 ordered by primary care physicians in a managed care organization in Israel through redesign of a computerized order form. STUDY DESIGN: Pre-post intervention. METHODS: A new version of the computerized order form was launched. Utilization patterns were calculated for tests of vitamin B12, folic acid, and ferritin, which were previously grouped together. Concomitant utilization patterns for tests of hemoglobin and iron were evaluated as controls. RESULTS: Tests ordered for the 3 targets decreased by 31% to 41% relative to the preintervention month, with a further decrease to 36% to 53% the following month. Negligible changes in utilization patterns were observed for the controls (-2% to 3%) during the postintervention period. CONCLUSIONS: Simple restructuring of a computerized order form significantly reduced the number of laboratory tests suspected of being unnecessary or redundant. When overutilization of laboratory resources is suspected, managers should evaluate the efficiency of the organization's current ordering procedures before implementing resource-intensive interventions. PMID- 19298099 TI - Knowledge, attitudes, and preferences regarding advance directives among patients of a managed care organization. AB - OBJECTIVES: To examine attitudes, experiences, and preferences regarding advance directives (ADs) among adults of all ages. STUDY DESIGN: Subjects were surveyed regarding end-of-life (EOL) care wishes. The survey included items measuring knowledge, experiences, and attitudes regarding ADs, as well as preferences toward initiation of discussions. METHODS: Subjects included a random sample of patients (age range, 20 to >80 years) from a large midwestern managed care organization stratified by age decade. Descriptive statistics were used to summarize variables, and chi2 tests were performed to examine differences by age category. RESULTS: One hundred eighty-seven surveys were completed. The likelihood of having completed an AD increased with age (P <.001). Of those without an AD, 44% reported having talked with someone about their wishes. Many (62%) subjects thought that it was up to them to raise the topic of EOL care. However, 70% of subjects reported that they would be comfortable if their provider initiated discussion. Participants 60 years or older were more likely to report being very comfortable with their provider bringing up the subject compared with those younger than 60 years (60% vs 39%, P = .02). Few differences were found by age. CONCLUSIONS: Room for improvement exists for increasing the number of patients who complete an AD or engage in discussion of their wishes. Ways to involve healthcare providers in the process should be explored, as it seems that patients are receptive to physician-initiated discussions of ADs. PMID- 19298100 TI - Medicare Part D coverage gap and diabetes beneficiaries. AB - OBJECTIVE: To examine drug costs and entry and exit rates into the Part D coverage gap for beneficiaries with diabetes in Medicare Advantage managed care plans. STUDY DESIGN: Cross-sectional observational study. METHODS: Study patients were Medicare Advantage Part D beneficiaries with diabetes from 2 large California health plans who were continuously enrolled in 2006 and had a drug coverage gap starting at $2250. Entry and exit into the gap, total drug costs, and out-of-pocket drug costs were determined using pharmacy databases. RESULTS: In 2006, 26% of the 42,801 beneficiaries with diabetes reached the coverage gap; 2% of beneficiaries exited the gap and qualified for catastrophic coverage. Beneficiaries incurred a mean of $2182 in total drug costs during 2006. Drug expenditures remained stable over the year for beneficiaries who did not enter the gap. For beneficiaries who entered the gap, total drug costs were higher overall and decreased at year's end as out-of-pocket expenses increased. CONCLUSIONS: Fewer diabetes patients in this study entered the coverage gap than had been previously estimated, but the entry rate was much higher than that of the general Medicare Advantage Part D population. Patients entering the gap had lower subsequent monthly drug expenditures; this may be due to lower-than expected drug prices and greater use of generics in managed care, or it may potentially signal poorer drug adherence. Future work should examine these hypotheses and explore risk factors for entering the Part D coverage gap. PMID- 19298101 TI - Patient factors associated with following a relocated primary care provider among older adults. AB - OBJECTIVES: To use a natural experiment to identify patient factors associated with the decision to follow one's primary care provider (PCP) to a more distant location after the closure of a medical practice. STUDY DESIGN: Case-control study. METHODS: Eight months after the closure of a practice in Dundalk, Maryland, we randomly selected 140 patients older than 60 years from each of the following groups: those who followed their PCP (continuity group) and those who transferred to a closer clinic (proximity group). We designed a survey instrument to collect information about demographics, duration of the patient-PCP relationship, transportation, self-assessed driving proficiency, and patients' estimates of the distance in miles and the driving time in minutes from their homes to both practices. Chi2 tests and logistic regression analyses were used to determine differences between the groups. RESULTS: The response rate to the survey was 64%. More than 85% of patients in both groups had been with their original PCP for longer than 2 years. In multivariable analysis, the following 3 factors were associated with being a patient in the proximity group: living alone (adjusted odds ratio [OR], 3.14; 95% confidence interval [CI], 1.35-7.26), having greater physical disability (physical component summary score <40; OR, 2.14; 95% CI, 1.04-4.39), and perceiving that travel time from home to the farther clinic would require at least 10 minutes longer than the MapQuest estimate (OR, 4.08; 95% CI, 1.97-8.43). CONCLUSION: Older patients who live alone and are weaker seem to be more likely to forgo continuity with their PCP for the sake of convenience when a barrier to access occurs such as relocation of the physician to a more distant office. PMID- 19298102 TI - Late-life comorbid insomnia: diagnosis and treatment. AB - Changing sleep architecture in the elderly may increase their vulnerability to comorbid insomnia. Common comorbid conditions include chronic pain, depression, nocturia, and neurologic conditions such as Parkinson's and Alzheimer's disease. Diagnosing and treating comorbid insomnia in an older population poses special challenges for clinicians given the variety of coexisting medical and psychological conditions, polypharmacy, and the potential adverse effects of the most commonly used medications for insomnia in this population. Thus, the use of nonpharmacologic treatments, such as cognitive behavior therapy and relaxation techniques, is recommended before any medical approaches. PMID- 19298103 TI - Current and new thinking in the management of comorbid insomnia. AB - Insomnia occurs predominantly in conjunction with a medical or psychiatric illness. New thinking regarding the treatment of comorbid insomnia has moved the field away from practices that called for treating the comorbid condition to resolve the coexisting insomnia to one in which the insomnia is treated as a separate condition. Although 10 medications currently are approved by the US Food and Drug Administration for the treatment of insomnia, only 2, eszopiclone and zolpidem, have been evaluated for efficacy in patients with chronic comorbid insomnia. Studies suggest clear benefits in comorbid insomnia. Nonpharmacologic treatments, such as cognitive behavioral therapy, sleep hygiene, and relaxation training, have also been investigated for comorbid insomnia, with studies suggesting these approaches may be effective either alone or in conjunction with medications. While behavioral issues should be optimized, clinicians need to customize treatments for patients with comorbid insomnia based on coexisting medical and psychiatric morbidities, age, medical history, current medications, and lifestyle issues. PMID- 19298104 TI - Comorbid insomnia: current directions and future challenges. AB - Insomnia is a leading cause of absenteeism, presenteeism (lost productivity when employees are at work), accidents, and errors in the workplace. Overall direct and indirect costs exceed $30 billion annually. A significant portion of these costs are attributable to patients with comorbid insomnia, making these conditions a significant clinical public health issue. These comorbid conditions include mood and anxiety disorders; chronic pain; respiratory, urinary, and neurologic conditions; diabetes; and cardiovascular diseases. Traditional treatment for insomnia with comorbid conditions has focused on treating the comorbid condition with the expectation that the insomnia will resolve. Recent studies, however, suggest this approach is not the most appropriate. Instead, treating both conditions simultaneously may improve the outcomes for each. PMID- 19298108 TI - Particle lung interactions. PMID- 19298110 TI - Editor's Note. PMID- 19298109 TI - Introduction. PMID- 19298111 TI - Consensus statement: aerosols and delivery devices. PMID- 19298112 TI - Plenary Lecture: INHALED DRUG DELIVERY, PAST, PRESENT AND FUTURE: A THERAPEUTIC PERSPECTIVE. PMID- 19298113 TI - Plenary Lecture: Site-Directed Aerosol Delivery of HFA-Beclomethasone Dipropionate to the Lungs. PMID- 19298115 TI - Clinical application-diagnosis. PMID- 19298114 TI - Inhalation delivery of therapeutic peptides and proteins. PMID- 19298116 TI - Clinical application-therapy. PMID- 19298118 TI - Particle-lung interactions: molecular effects. PMID- 19298117 TI - Particle-lung interactions: cellular effects. PMID- 19298119 TI - Environmental aerosols. PMID- 19298120 TI - Therapeutic aerosols. PMID- 19298121 TI - Therapeutic aspects. PMID- 19298122 TI - Technical aspects. PMID- 19298123 TI - Formulation. PMID- 19298125 TI - Standardization. PMID- 19298124 TI - Modeling. PMID- 19298127 TI - Mucus and cilia symposium. PMID- 19298126 TI - Aerosol delivery systems. PMID- 19298128 TI - Aerosols for Medical Applications (GAeF Satellite Meeting). PMID- 19298129 TI - ASTRA Satellite Symposium -"The Importance of Size". PMID- 19298131 TI - Partial rescue of growth failure in growth hormone (GH)-deficient mice by a single injection of a double-stranded adeno-associated viral vector expressing the GH gene driven by a muscle-specific regulatory cassette. AB - Growth hormone (GH) deficiency (GHD) causes somatic growth impairment. GH has a short half-life and therefore it must be administered by daily subcutaneous injections. Adeno-associated viral (AAV) vectors have been used to deliver genes to animals, and double-stranded AAV (dsAAV) vectors provide widespread and stable transgene expression. In the present study we tested whether an intramuscular injection of dsAAV vector expressing GH under the control of a muscle creatine kinase regulatory cassette would ensure sufficient systemic GH delivery in conjunction with muscle-specific expression. Virus-injected GHD mice showed a significant (p < 0.05) increase in body length and body weight, without reaching full normalization, and significant (p < 0.05) reduction in absolute and relative visceral fat. Quantitative RT-PCR showed preferential GH expression in skeletal muscles that was confirmed by qualitative fluorescence analysis in mice injected with a similar virus expressing green fluorescent protein. The present study shows that systemic GH delivery to GHD animals is possible via a single intramuscular injection of dsAAV carrying a muscle-specific GH-expressing regulatory cassette. PMID- 19298133 TI - Forensic pregnancy testing: a special case in molecular diagnostics. PMID- 19298132 TI - The oncopathic potency of Clostridium perfringens is independent of its alpha toxin gene. AB - Hypoxia in solid tumors is a major obstacle in conventional treatment because of inefficient delivery of therapeutic agents to the lesions, but offers the potential for anaerobic bacterial colonization that can lead to tumor destruction. We have previously reported a recombinant Clostridium perfringens (Cp) strain constructed by deletion of the superoxide dismutase (sod) gene and insertion of the Panton-Valentine leukocidin (PVL) gene, Cp/sod(-)/PVL, which showed elevated oxygen sensitivity, tumor selectivity, and oncopathic potency in an orthotopic model of pancreatic cancer in immune-competent and syngeneic mice, and that led to substantial prolongation of animal survival. A major limitation to Cp/sod(-)/PVL in clinical applications is that it expresses phospholipase C (plc), the alpha-toxin and the major virulence determinant in Cp that is causative in the development of gas gangrene. In this study, the plc gene in Cp/sod(-)/PVL was knocked out to create Cp/plc(-)/sod(-)/PVL, which was shown to be incapable of inducing gas gangrene in mice. Intravenous injection of Cp/plc( )/sod(-)/PVL spores led to a significant survival advantage in tumor-bearing mice with the same efficacy as Cp/sod(-)/PVL, indicating that the oncopathic potency of Cp is independent of a functional plc gene. The treatment also did not lead to an attenuated immune response to a subsequent pathogen challenge, indicating that a systemic immune-suppressive effect in the host is absent. Consequently, Cp/plc( )/sod(-)/PVL is a novel oncopathic bacterial agent for the effective treatment of pancreatic cancer and other poorly vascularized tumors, with a substantially enhanced safety profile, which is essential for the development of translational studies in the future. PMID- 19298134 TI - Spermatogonial stem cell biomarkers: improved outcomes of spermatogonial transplantation in male fertility restoration? PMID- 19298135 TI - Circulating tumor cells: utility for predicting response to anti-EGFR therapies? PMID- 19298137 TI - Endoscopy, morphology, morphometry and molecular markers: predicting cancer risk in colorectal adenoma. AB - The evaluation of short- and long-term risk for developing cancer in patients with colorectal adenomas is controversial. Good, reliable predictors of cancer risk in any adenoma are currently lacking and are limited to adenoma size, number and histologic type. In fact, the evaluation of any adenoma or precancer lesion (e.g., hyperplastic polyps, serrated adenoma or aberrant crypt foci) within the colorectum may be assessed by a number of techniques ranging from direct visualization through the endoscope, to microscopic assessment, and to evaluation at the molecular level. Emerging techniques may yield improved methods of adenoma risk-assessment in the near future. For one, newer endoscopy technologies include chromoendoscopy or endocytoscopy, which now render endoscopists able to resolve the surface and subsurface mucosa at cellular resolution in vivo and in real time - thus, bringing the microscope to the patient's bedside. This new era in endoscopic imaging is dubbed 'histoendoscopy'. Further, while traditional views of classifying protruding and sessile lesions include those of Haggitt, the sm classification, the Japanese and the so-called Vienna classifications to evaluate neoplasia, the development of new molecular techniques may give way to new methods of classifying preneoplasia and precancerous lesions. This review discusses some pros and cons of risk evaluation technologies in the colorectal tract by endoscopy, microscopy, and quantitative and molecular features. The morphometry-based studies performed over the past decades for the quantitative assessment of cellular and nuclear features within adenomas have failed to yield results amenable for clinical translation and are unlikely to improve further and gain widespread use with current technology. Rather, emerging knowledge of pathway-specific markers through the outlining of a molecular classification will likely be the basis for improved detection and diagnosis. The emerging genomic and proteomic technologies allowing for noninvasive tests to detect (asymptomatic) cancer and neoplasia are discussed. Lastly, the importance of recognizing bias and pitfalls and the adherence to guidelines for biomarker research are addressed. PMID- 19298138 TI - Immune-mediated liver diseases: programmed cell death ligands and circulating apoptotic markers. AB - Primary biliary cirrhosis, primary sclerosing cholangitis and autoimmune hepatitis are the three major immune-mediated liver diseases. The etiologies of primary biliary cirrhosis, primary sclerosing cholangitis and autoimmune hepatitis are largely unknown, but seem to be influenced by genetic and environmental factors. Autoantibodies can be found in nearly all patients with primary sclerosing cholangitis and autoimmune hepatitis, and in the vast majority of patients with primary sclerosing cholangitis. In addition, autoimmune hepatitis is associated with high concentrations of serum globulins. Enhanced liver cell death by apoptosis has been described in all of these liver diseases, although the precise mechanisms remain unclear. In general, apoptosis can be initiated via an extrinsic pathway that is triggered by engagement of death receptors on the cell surface, or via an intrinsic pathway that is induced by mitochondrial injury and is influenced by members of the Bcl-2 family. In both pathways, effector caspases are finally activated that cleave and degrade cell structures, resulting in the release of apoptotic products into the circulation. New diagnostic tests can detect these apoptotic markers and programmed cell death ligands such as Fas and Fas-ligands, nucleosomes, caspases, cytokeratin fragments, macrophage migration inhibitory factor, soluble intracellular adhesion molecule, natural killer cells group 2D and programmed death ligands. Several of these markers have been found to be altered in tissue and/or blood of immune mediated liver diseases, some also in nonimmune-mediated liver diseases. Beyond their potential usefulness as additional diagnostic markers, they may be valuable for the estimation of disease severity and therapy monitoring. This review summarizes current knowledge on apoptotic mechanisms, death receptor ligands and circulating apoptotic markers in immune-mediated liver diseases. PMID- 19298139 TI - Allergic bronchopulmonary aspergillosis: the hunt for a diagnostic serological marker in cystic fibrosis patients. AB - The diagnosis of allergic bronchopulmonary aspergillosis (ABPA) in cystic fibrosis patients remains challenging, mainly owing to overlapping symptoms of the underlying lung disease with clinical symptoms of ABPA. In addition, a varying mixture of diagnostic criteria, including clinical status, radiological findings and immunological measurements, has led to confusion and differing recommendations. In order to help simplify as well as standardize the diagnostic criteria for ABPA, different serological markers have been evaluated in the last 20 years and their usefulness has been assessed in many clinical studies. This review presents current diagnostic criteria of ABPA, with a special focus on serum markers supporting the diagnosis and explains why the hunt for a serological marker for ABPA is still ongoing. PMID- 19298140 TI - IFN-gamma release assays in tuberculosis management in selected high-risk populations. AB - Tuberculosis (TB) is the most deadly infectious disease in the world. TB control relies on passive case findings and targeted treatment of latently infected individuals at high risk of disease progression. Tuberculin skin testing (TST) is conventionally used for detection of TB infection. Recently, blood assays measuring the release of IFN-gamma by TB-specific effector memory T cells have been developed to overcome TST limitations. Overall, IFN-gamma release assays are more specific than TST, more sensitive in detecting active TB and correlate better with TB exposure in immune-competent patients, at least in low-burden settings. There are three US FDA-approved assays commercially available: the ELISpot-based assay T-SPOT.TB (Oxford Immunotech, UK) and two ELISA-based formats, QuantiFERON TB Gold (QFT) and QFT-in tube (Cellestis, Australia). Recent international guidelines and consensus statements recommend the use of IFN-gamma release assays at different levels in TB management. However, conclusive evidence based information targeting populations at high TB risk, including HIV-infected individuals, children and patient candidates for biotherapy with TNF-alpha blockers, are lacking. The aim of this review is to focus our attention on studies addressing the performance of commercial IFN-gamma release assays in clinical management of TB infection in these highly selected settings to provide a more comprehensive picture of the actual scenario and to identify areas to be investigated further. PMID- 19298141 TI - Diagnostic and analytical mutation scanning of Cryptosporidium: utility and advantages. AB - Cryptosporidiosis is predominantly a disease of the alimentary tract of humans and other vertebrates, caused by parasitic protists of the genus Cryptosporidium. This disease, transmitted mainly via the fecal-oral route (in water or food), is of major socioeconomic importance globally. The diagnosis of cryptosporidiosis, including the genetic characterization of the different species, genotypes and subgenotypes (population variants) of Cryptosporidium, is crucial to prevention and control, particularly as there is no cost-effective treatment against this disease. Although traditional phenetic techniques have had major deficiencies for the specific diagnosis of cryptosporidiosis, there has been substantial progress in the establishment of molecular tools. In this article, we review key genetic markers used for the specific identification of Cryptosporidium, diagnosis of cryptosporidiosis and analysis of genetic variation in Cryptosporidium populations. We also discuss the advantages and disadvantages of selected techniques, and emphasize the benefits of utilizing rapid mutation scanning in achieving improved insights into the population genetics and epidemiology of Cryptosporidium. PMID- 19298142 TI - Reliability of real-time reverse-transcription PCR in clinical diagnostics: gold standard or substandard? AB - Molecular diagnostics is one of the major growth areas of modern medicine, with real-time PCR established as a qualitative and quantitative technology that is rapid, accurate and sensitive. The sequencing of the human genome, comprehensive genomic, mRNA and miRNA expression profiling of numerous cancer types, the ongoing identification of disease-associated polymorphisms and the expanding availability of genomic sequence information for human pathogens has opened the door to a wide range of translational applications for this technology. Consequently, novel real-time PCR assays have been developed for diagnosis and prognosis, treatment monitoring, transplant biology and pathogen detection, as well as more controversial uses such as lifestyle genotyping. However, this technology is still troubled by significant technical deficiencies. Hence its often-improper use as a clinical tool has important public health implications, most recently demonstrated through its association with the measles, mumps and rubella vaccine/autism controversy. This serves as a timely reminder of the indispensable requirement for careful experimental design, validation and analysis. PMID- 19298144 TI - Suicide and life-threatening behavior. In this issue. PMID- 19298145 TI - Empirical support for an evolutionary model of self-destructive motivation. AB - We tested predictions generated from an evolutionary account of self-destructive motivation in two survey studies of 18-24-year-old university students. As hypothesized, hierarchical regressions showed that the positive relationship between perceived burden to family and suicide ideation was amplified for participants with low measured health and romantic relationship satisfaction, and for participants with relatively young mothers. The moderating effect of maternal age was also observed in logistic regressions of suicide attempts. These effects occurred independently of depression, hopelessness, and other relevant extraneous variables. Results have implications for understanding self-destructive motivation, assessing suicide risk, and preventing suicidal thinking and behavior. PMID- 19298146 TI - Gender and suicide risk: the role of wound site. AB - That males have higher suicide rates than females is one of the most empirically documented social facts in suicidology, but the reasons for this continue to be debated. For the present paper, we tested a neglected contributing factor to the gender suicide ratio: wound site or the area of the body that is wounded in firearm suicides. Males may have a higher suicide rate, in part, due to their greater likelihood than women for shooting themselves in the head as opposed to the body. This has been related to gender differences in fear of facial disfigurement and suicide intent. Data from the Wayne County Medical examiner's office regarding 807 suicides committed with firearms was analyzed. The dependent variable was dichotomous and referred to the location of the site of the wound: gunshot to the head vs. gunshot to the body. Controls for demographic covariates of suicide included age and race of the suicide victim. The results of the multivariate logistic regression analysis determined that women were 47% less apt than men to shoot themselves in the head. Further analysis determined that women were less apt than men to use shotguns and rifles in their suicides (weapons that make head shooting more awkward). The findings are consistent with the notion that women are more concerned than men with facial disfigurement, and that women have a lower desire to die than men. PMID- 19298147 TI - Training mental health professionals to assess and manage suicidal behavior: can provider confidence and practice behaviors be altered? AB - Remarkably little systematic research has studied the effects of clinical suicidology training on changing practitioner attitudes and behaviors. In the current study we investigated whether training in an empirically-based assessment and treatment approach to suicidal patients administered through a continuing education workshop could meaningfully impact professional practices, clinic policy, clinician confidence, and beliefs posttraining and 6 months later. At the 6 month follow-up we found that 44% of practitioners reported increased confidence in assessing suicide risk, 54% reported increased confidence in managing suicidal patients, 83% reported changing suicide care practices, and 66% reported changing clinic policy. These results suggest that a brief and carefully developed workshop training experience can potentially change provider perceptions and behaviors with a possible impact on clinical care therein. PMID- 19298148 TI - The benefit of forgetting suicidal ideation. AB - In a sample of young adult Australians, those who had had suicidal ideation but who did not acknowledge ever having had it when asked 4 years later, were experiencing better mental health, as demonstrated by significantly better functioning on a range of psychometric measures, than those who recalled it. These results are consistent with several recent reports and indicate that forgetting painful events such as suicidal ideation is an adaptive defense mechanism. This has implications in terms of therapy focusing on contemporaneous events and the future, rather than on the past. PMID- 19298149 TI - Assessment of depression and suicidal actions: agreement between suicide attempters and informant reports. AB - Knowledgeable informants may be able to provide useful information about depressive symptoms and suicidal actions when a suicidal patient is uncooperative with a clinical interview or not available for a psychiatric evaluation. The present study was designed to examine information gathered from psychiatric inpatients who had attempted suicide as compared to similar information obtained from a close friend or family member. From a larger sample of adult psychiatric inpatients diagnosed with major depression, 25 suicide attempters were assessed and their reports were compared to similar information solicited from a close friend or family member. The assessment focused on depressive symptoms from the SCID and the presence of various suicidal actions. Moderate to high agreement was found on all measures. Psychiatric patients and their informants tended to agree on the presence of depressive symptoms, the presence of recent stressful life events, and specific actions taken during the suicidal crisis, suggesting that informants may be capable of providing useful information about the patient. PMID- 19298150 TI - Correlates of serious suicidal ideation and attempts in female adult sexual assault survivors. AB - Relations between (a) serious suicidal ideation and attempts and (b) demographics, trauma history, assault characteristics, post-assault outcomes, and psychosocial variables were examined among female adult sexual assault survivors. Younger, minority, and bisexual survivors reported greater ideation. More traumas, drug use, and assault disclosure related to greater attempts, whereas perceived control over recovery was related to fewer attempts. Child sexual abuse and some assault characteristics predicted suicidal behavior. Depression was related to suicidal behavior until psychosocial variables were accounted for. Specifically, using substances to cope and self-blame predicted greater ideation, whereas receiving aid/information support was related to less ideation. Implications for research and treatment are discussed. PMID- 19298151 TI - The role of body image and disordered eating as risk factors for depression and suicidal ideation in adolescents. AB - There is much empirical literature on factors for adolescent suicide risk, but body image and disordered eating are rarely included in these models. In the current study, disordered eating and body image were examined as risk factors for suicide ideation since these factors are prevalent in adolescence, particularly for females. It was hypothesized that disordered eating and body image, in addition to depressive symptoms, would contribute to suicide ideation. It was also hypothesized that these relationships would be stronger for females than for males. Structural equation modeling was used to test a model of risk for suicide ideation incorporating the above factors in a sample of 392 high school students. Results indicated that disordered eating contributed to both suicide ideation and depressive symptoms, while body image only contributed to depressive symptoms. Depressive symptoms contributed to suicide ideation. The model was found to be cross-validated with males and females, and no gender differences emerged. Implications of these findings and their importance in constructing future models of adolescent suicide risk are discussed. PMID- 19298152 TI - Suicidal behavior in the Western Pacific region: characteristics and trends. AB - Current knowledge of suicidal behavior in the Western Pacific region is relatively limited when compared to other areas of the world. This area is characterized by a wide diversity of social, economic, and cultural aspects that do not permit any homogenous reading of suicide phenomena. Data from countries of the region included in this article suggest the existence of important differences among nations in terms of gender, age, and methods of suicide, as well role of mental illness. In addition, the Western Pacific region appears to be affected by higher rates of suicide than other areas in the world, with signs of progressive worsening in recent years and worrying predictions for the next two decades. The Suicide Trends in At-Risk Territories (START) Study, initiated by the World Health Organization Regional Office in Manila, seeks to research the incidence of both fatal and nonfatal suicidal behavior in countries of the region and to implement interventions for suicide attempters. This ambitious project is expected to standardize data collection procedures and improve the knowledge cultural factors in suicide. PMID- 19298153 TI - An integrative suicide prevention program for visitor charcoal burning suicide and suicide pact. AB - An integrative suicide prevention program was implemented to tackle an outbreak of visitor charcoal burning suicides in Cheung Chau, an island in Hong Kong, in 2002. This study evaluated the effectiveness of the program. The numbers of visitor suicides reduced from 37 deaths in the 51 months prior to program implementation to 6 deaths in the 42 months post-implementation period. The number of visitor suicide pacts decreased from 7 pacts (15 individuals) to 1 pact (2 individuals). No statistically significant differences in the numbers of visitor suicide attempts and resident suicides were observed in the two time periods. No statistically significant changes in visitor suicides during the study period were observed on the comparison islands. The consistency and timing of reduction in visitor suicides correlated with the development and delivery of the integrative program on the intervention island, suggesting a causal association between program delivery and reduction of visitor suicides. The possibility of displacement seems small because there was no increase in visitor suicides on the comparison islands during the study period. This integrative approach in preventing target-specific suicides may serve as an example for other communities to develop suicide prevention programs that make use of the existing local resources. PMID- 19298154 TI - Suicide and its association with individual, family, peer, and school factors in an adolescent population in southern Taiwan. AB - A representative sample of 10,233 adolescent students was recruited to examine the rate of suicidal attempt and its correlates in the adolescents living in southern Taiwan. Five questions from the Kiddie Schedule for Affective Disorders and Schizophrenia (Kiddie-SADS-E) were used to inquire about the participants' suicidality. The associations between suicidal attempt and multidimensional factors were examined by using logistic regression analysis: 9.1% of the participants reported a suicidal attempt in the preceding year. Female gender, low self-esteem, weekly alcohol use, illicit drugs use, depression, high family conflict, low maternal education level, poor family function, low connectedness to school, low rank, poor feeling in peer group, and drop out from school were associated with adolescent suicidal attempt. The rate of suicidal attempt was found to be high in Taiwanese adolescents, and multidimensional factors were correlated to adolescent suicidal attempt. PMID- 19298155 TI - Epidemiologic characteristics and trends of fatal suicides among the elderly in Taiwan. AB - Taiwan has one of the highest suicide rates in the world, especially among its elderly. The epidemiologic characteristics and trends of the surging elderly suicide rates from 1993 to 2003 are described, with a special emphasis on the risk groups, the methods used in suicide, and their geographical variations. Data on annual mortality for persons over 65 years of age with external cause-of-death codes E950-E959 were obtained from the Death Certification data file provided by the Department of Health and used in the analysis. The suicide rate for each 5 year age group over 65 years old increased during the 11-year period. Elderly males had the highest suicide rate and experienced an increased rate of 49% during the decade. The average suicide ratio between elderly males and females was about 2:1. Never-married males had the highest age-adjusted as well as sex- and marital status-specific suicide rates, and showed an increased rate of 66% over the 11-year period. Geographical variations in suicide rates were significant, with the lest urbanized eastern Taiwan having a higher rate than other regions. Hanging, strangulation, and suffocation were the most used methods for committing suicide by the elderly, but their use had decreased from 63% to 54% during the decade; but jumping off a building and drowning increased significantly. Variation in suicide rate among months was not significant. With the increase in the elderly suicide rate, more governmental and societal interventions are needed to alleviate this social and human problem. PMID- 19298156 TI - Europe's advanced therapy medicinal products: chances and challenges. PMID- 19298158 TI - FlexPen for the delivery of insulin: accuracy, injection force and patient preference. AB - FlexPen is a widely used, prefilled, insulin-delivery pen for patients with Type 1 or Type 2 diabetes. Well-designed trials suggest that FlexPen is more accurate than other prefilled pens at high, medium and low doses. Furthermore, FlexPen is preferred to the vial-and-syringe method by patients, but comparisons of patient preference for modern pens are currently lacking. Next Generation FlexPen has similar accuracy to FlexPen but has improved patient perception. FlexPen improves adherence and reduces costs compared with the vial-and-syringe method, but the relative adherence and/or cost benefits of different pens are unknown. Important improvements in Next Generation FlexPen include: a significantly reduced injection force (that is lower than the injection force of other prefilled pens) and clearer labeling for insulin-type differentiation. Current evidence suggests that FlexPen and Next Generation FlexPen are an attractive choice when considering insulin-delivery methods. PMID- 19298159 TI - Masimo Rad-57 Pulse CO-Oximeter for noninvasive carboxyhemoglobin measurement. AB - Noninvasive methods of body fluid chemical measurement have been expanding. New technologies are enabling the quantification of different compounds in the blood and interstitial tissues. One example of this is the pulse oximeter, which has facilitated the measurement of oxyhemoglobin rapidly and reliably without the requirement of blood-draws. The Masimo Rad-57 Pulse CO-Oximeter expanded the capabilities of pulse-oximetry to include measurements of carboxyhemoglobin and methemoglobin. This innovation has revolutionized the paradigm for detection of patients with CO poisoning. Previously, clinicians relied on historical information and patient signs and symptoms pointing to the possibility of CO exposure or toxicity. Only then would a blood test be ordered to measure carboxyhemoglobin levels. Since the presentation of CO poisoning is nonspecific and overlaps with many other conditions, and since the presence of environmental CO is often unknown, the detection of this condition was only possible in cases where the presence of CO was obvious or where the symptoms were severe. We now know, from studies conducted using the Rad-57, the only US FDA-approved device for noninvasive measurement of SpCO, that there are a significant number of patients who experience CO exposure but are nonsymptomatic. The Rad-57 provides a clinical justification for screening in the healthcare setting to identify patients with significant CO exposure who would otherwise be undetected. PMID- 19298160 TI - Device safety and effectiveness in the pediatric population: a US FDA perspective. AB - The US FDA recognizes that there are many challenges to ensuring that medical devices are used responsibly and safely in the pediatric population. Education plays a key role, and manufacturers and healthcare providers must be informed of medical device issues unique to the pediatric population. Healthcare workers and device manufacturers must recognize that adverse events may differ between the adult and pediatric population and among pediatric subpopulations. Risk mitigation may also differ, and heightened awareness will reduce the potential for adverse events and promote the safe use of medical devices in children. PMID- 19298161 TI - Humanitarian Use Device and Humanitarian Device Exemption regulatory programs: pros and cons. AB - The US FDA established the Humanitarian Use Device (HUD) and Humanitarian Device Exemption (HDE) program to encourage medical device firms to address rare diseases. Despite being in existence for over a decade, there has only been one peer-reviewed publication examining this field. The objective of this report is to investigate how the HUD/HDE program differs from the standard regulatory system, discuss its potential advantages and disadvantages, and to speculate which humanitarian devices will be brought to market within the next 5 years. A total of 40 semistructured interviews with stakeholders, representing approximately half (n = 20, 49%) of the firms that have successfully obtained HDE approved products, were performed in order to acquire the primary data for this paper. There appear to be short-term gains and long-term drains associated with launching humanitarian devices to market. This report aims to provide sponsors with information that may allow them to make better decisions during their product development of humanitarian devices and may, hopefully, also play a role in encouraging other sponsors to take the necessary steps forward in helping to find treatments for patients with rare diseases. PMID- 19298162 TI - Oral inhalation therapy: meeting the challenge of developing more patient appropriate devices. AB - Although oral inhalers have been mass produced for more than 50 years, there is a large body of literature in which evidence has been provided that patients either misuse their inhalers inadvertently or deliberately, thereby reducing their intended efficacy or, in the worst cases, rendering them altogether ineffective. In general, inhalers are becoming increasingly complicated with the incorporation of add-on devices, miniaturized electronics and ever more complex mechanical systems that aid aerosol delivery to the lower respiratory tract and, at the same time provide user feedback. However, these benefits often come at a significant cost, and there are signs that increasing attention will need to be given to the cost-benefit equation in the future. This review explores the development of pressurized metered-dose inhalers, dry powder inhalers and devices for liquid droplet dispersal and inhalation from the perspective of the patient, by focusing on aspects that improve user interaction. These include designed-in features, such as breath-enhanced or breath-actuated operation that interact with the breathing pattern of the user, as well as more direct feedback aids that confirm, to the patient or healthcare provider that the dose has been delivered and that the patient has inhaled. PMID- 19298163 TI - Intracoronary optical coherence tomography and the evaluation of stents. AB - The application of optical coherence tomography to the cardiovascular system has led to intense interest, both clinically and also on the research front. Based on sound optical physics, intracoronary, in vivo imaging with high resolution (approximately 15 microm) has given unique insights into not only atherosclerotic plaques but also the tissue responses underlying stent implantation. This article will introduce the application of optical coherence tomography technology to coronary arteries, with particular reference to the evaluation of stents, and will also give a glimpse into current and future developments that will see optical coherence tomography remain a key tool in the armamentarium of researchers and interventional cardiologists alike. PMID- 19298164 TI - Anatomic and physiologic rationale for various technologies for primary total hip arthroplasty. AB - A number of patients have anatomic or physiologic variations that may adversely affect the performance of a primary total hip arthroplasty. Various technologies have been utilized in an attempt to improve the outcomes for these patients; however, some of these potential solutions are controversial. The authors examined the complete body of literature for scientific evidence regarding the use of these new technologies. The anatomic and physiologic anomalies that were studied include extra-articular deformities, developmental dysplasia, Perthes disease, Type C femoral bone, acetabular bone deficiency, femoral rotational abnormalities, variations that increase the risk of hip dislocation, sickle cell anemia, and extremely small or large bone sizes. This article presents the current scientific evidence and imparts an unbiased view of the use of various technologies to provide individualized solutions for patients who have anatomic or physiologic variations. PMID- 19298165 TI - Guidewires in clinical practice: applications and troubleshooting. AB - Guidewires are commonly used in clinical practice over a wide range of specialties. Their use has become more popular as a result of advances in endourology and interventional radiology, as well as in angiographic procedures. While there are many papers regarding individual guidewires and certain technical aspects, for the interested clinician there is little to give a generalized overview within the literature. This paper aims to review guidewires in terms of their make-up, applications and potential complications. Technical points are described, as well as rescue methods for complications, in the hope of preventing future litigation for those reading the paper. PMID- 19298166 TI - Diffusion of medical technology: medical devices in India. AB - This review examines the diffusion of modern medical devices in India by analyzing trends in India's cross-border trade in medical devices, its domestic medical device production and utilization by households. We explore the implications of this process of diffusion for the efficacy, cost-effectiveness and equitable use of new medical devices in India, and review recent efforts to regulate the Indian medical device sector. PMID- 19298168 TI - Culture conditions allow selection of different mesenchymal progenitors from adult mouse bone marrow. AB - The use of adult stem cells in tissue engineering approaches will benefit from the establishment of culture conditions that allow the expansion and maintenance of cells with stem cell-like activity and high differentiation potential. In the field of adult stem cells, bone marrow stromal cells (BMSCs) are promising candidates. In the present study, we define, for the first time, conditions for optimizing the yields of cultures enriched for specific progenitors of bone marrow. Using four distinct culture conditions, supernatants from culture of bone fragments, marrow stroma cell line MS-5, embryonic fibroblast cell line NIH3T3, and a cocktail of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF), we isolated four different sub-populations of murine BMSCs (mBMSCs). These cells express a well-known marker of undifferentiated embryonic stem cells (Nanog) and show interesting features in immunophenotype, self-renewal ability, and differentiation potency. In particular, using NIH3T3 conditioned medium, we obtained cells that showed impairment in osteogenic and chondrogenic differentiation while retaining high adipogenic potential during passages. Our results indicate that the choice of the medium used for isolation and expansion of mBMSCs is important for enriching the culture of desired specific progenitors. PMID- 19298170 TI - Trends in postoperative sepsis: are we improving outcomes? AB - BACKGROUND AND PURPOSE: Each year, as many as two million operations are complicated by surgical site infections in the United States, and surgical patients account for 30% of patients with sepsis. The purpose of this study was to determine recent trends in sepsis incidence, severity, and mortality rate after surgical procedures and to evaluate changes in the pattern of septicemia pathogens over time. METHODS: Analysis of the 1990-2006 hospital discharge data from the Healthcare Cost and Utilization Project (HCUP) State Inpatient Databases (SID) for New Jersey. Patients >or= 18 years who developed sepsis after surgery were identified using International Classification of Diseases, Ninth Revision, Clinical Modification codes as defined by the Patient Safety Indicator "Postoperative Sepsis" developed by the Agency for Healthcare Research and Quality (AHRQ). Severe sepsis was defined as sepsis complicated by organ dysfunction. RESULTS: A total of 1,276,451 surgery discharges (537,843 elective [42.1%] and 738,608 non-elective [57.9%] procedures) were identified. After elective surgery, 5,865 patients (1.09%) developed postoperative sepsis, of whom 2,778 (0.52%) had severe sepsis. The incidence of postoperative sepsis after elective surgery increased from 0.67% to 1.74% (p < 0.0001) and severe sepsis after elective surgery from 0.22% to 1.12% (p < 0.0001). The sepsis mortality rate for elective procedures showed no significant change over time. The proportion of severe sepsis after elective cases increased from 32.9% to 64.6% (p < 0.0002). The rates of postoperative sepsis (4.24%) and severe sepsis (2.28%) were significantly greater for non-elective than for elective procedures (p < 0.0002). Non-elective surgical procedures had a significant increase in the rates of postoperative sepsis (3.74% to 4.51%) and severe sepsis (1.79% to 3.15%) over time (p < 0.0001) with the proportion of severe sepsis increasing from 47.7% to 69.9% (p < 0.0002). The in-hospital mortality rate after non-elective surgery decreased from 37.9% to 29.8% (p < 0.0001). CONCLUSIONS: Sepsis and death were more likely after non-elective than elective surgery. Sepsis and severe sepsis has increased significantly after elective and non-elective procedures over the last 17 years. The hospital mortality rate was reduced significantly after non elective surgery, but no improvements were found for elective surgery patients who developed sepsis. Disparities in age, sex, and ethnicity and the development of postoperative surgical sepsis were found. Population-based studies may assist in defining temporal trends, disparities, and outcomes in sepsis not elucidated in smaller studies. PMID- 19298171 TI - Microbiology of infected acellular dermal matrix (AlloDerm) in patients requiring complex abdominal closure after emergency surgery. AB - BACKGROUND: Acellular dermal matrix (AlloDerm) has recently been introduced as an option for complex abdominal closure for patients with loss of abdominal wall domain secondary to intra-abdominal sepsis or necrotizing fasciitis. AlloDerm has been touted as a promoter of neovascularization and collagen deposition. Currently, the rate of AlloDerm infection in contaminated cases is unknown. Our objective was to determine if the organisms cultured during source control would infect AlloDerm. METHODS: The medical records of patients who required complex abdominal closure with AlloDerm in a tertiary-care hospital were reviewed from January to December, 2005. For each patient demographic, the reason for urgent surgery, American Society of Anesthesiologists (ASA) class, Acute Physiology and Chronic Health Evaluation (APACHE) II score, serum albumin concentration, culture results of purulent fluid obtained during surgery, and culture results of biopsies of infected-appearing AlloDerm (change of color, delayed granulation, odor) were collected. Data are presented as mean +/- standard error of the mean. RESULTS: Seventeen patients required the use of AlloDerm for tension-free closure of the abdominal wall after surgery for source control in necrotizing fasciitis (13%) or intra-abdominal sepsis (87%). The mean age was 61 +/- 2 years; 73% of the patients were Caucasian, the remainder being African American. The mean APACHE II score was 23.7 +/- 2.0, and the median ASA class was 3. The mean preoperative albumin concentration was 2.27 +/- 0.26 g/dL. Most (76%) of the patients had a wound vacuum-assisted closure system placed over the AlloDerm. Four patients (24%) were noted to have an infection of the AlloDerm graft at 24 +/- 10 days postoperatively. The cultures obtained at operation and from infected AlloDerm show similar organisms (Pseudomonas in two, Escherichia coli and methicillin-resistant Staphylococcus aureus in one each). Infected AlloDerm was coated with silver sulfadiazene and moistened dressings, and all four patients had complete resolution of the AlloDerm infection with an adequate bed of granulation tissue, allowing skin grafting. CONCLUSION: Patients with contaminated abdomens who require complex closure with AlloDerm are at risk of developing infection of their graft material with organisms similar to those present at the time of surgery. Once culture results are obtained, topical antimicrobials with activity against the cultured organisms may be employed as part of the AlloDerm dressings to prevent infection and promote healing. PMID- 19298172 TI - Monthly screening for polyoma virus eliminates BK nephropathy and preserves renal function. AB - BACKGROUND: Polyoma BK virus nephropathy is a serious complication after renal transplantation and is associated with a high rate of allograft failure. Progressive infection with BK virus in immunocompromised renal transplant recipients occurs in detectable stages: Viruria, viremia, then nephropathy. METHODS: In January, 2006, we initiated a plasma screening policy for all new transplant recipients, with monthly blood testing for BK virus by polymerase chain reaction (PCR). Between January 1, 2006, and February 28, 2007, 66 renal transplants were performed at our center. The 11 patients with a positive plasma BK PCR test underwent prompt reduction in baseline immunotherapy consisting of a 50% daily dose reduction (n = 6) or complete discontinuation of therapy with mycophenolate mofetil (n = 5). RESULTS: After reduction or discontinuation of mycophenolate mofetil, 10 patients became negative for BK virus in the plasma within 6 months. Progression to BK nephropathy has not occurred, and renal transplant dysfunction secondary to acute cellular rejection developed in only 1 patient (9%). One year post-transplant, the mean serum creatinine values for these 11 patients remained stable at 1.5 mg/dL. CONCLUSION: Monthly plasma screening for BK virus by PCR together with immunosuppressive regimen reduction prevents BK nephropathy. In addition, this intensive screening protocol is associated with a low rate of acute rejection and excellent preservation of renal function. PMID- 19298173 TI - Lentiviral delivery of LMX1a enhances dopaminergic phenotype in differentiated human bone marrow mesenchymal stem cells. AB - Human mesenchymal stem cells (MSCs) reside in the bone marrow and are known for their ability to differentiate along the mesenchymal lineage (fat, bone, and cartilage). Recent works have suggested the possibility that these cells are also capable of differentiating toward the neuroectodermal lineage. Using lentiviral gene delivery, we sought to reprogram the bone marrow-derived MSCs toward dopaminergic differentiation through delivery of LMX1a, which was reported to be a key player in dopaminergic differentiation in both developmental animal models and embryonic stem cells. Transduction of cells with fluorescent reporter genes confirmed efficiency of gene delivery. On incubation of the LMX1a transduced cells in differentiation medium, the LMX1a protein was concentrated in the cells' nuclei and specific dopaminergic developmental genes were upregulated. Moreover, the transduced cells expressed higher levels of tyrosine hydroxylase, the rate limiting enzyme in dopamine synthesis, and secreted significantly higher level of dopamine in comparison to nontransduced cells. We hereby present a novel strategy to facilitate the dopaminergic differentiation of bone marrow-derived MSCs as a possible cell source for autologous transplantation for Parkinsonian patients in the future. PMID- 19298174 TI - Human GPM6A is associated with differentiation and neuronal migration of neurons derived from human embryonic stem cells. AB - Glycoprotein M6A (GPM6A) is known as a transmembrane protein and an abundant cell surface protein on neurons in the central nervous system (CNS). However, the function of GPM6A in the differentiation of neurons derived from human embryonic stem (ES) cells is unknown. To investigate the function of GPM6A in neural differentiation, we generated human ES cell lines with overexpressed (B2h-oeM6A) or suppressed (B2h-shM6A) human GPM6A. Real-time polymerase chain reaction (PCR) showed that overexpression of GPM6A markedly increased the expression of neuroectodermal-associated genes (OTX1, Lmx1b, En1, Pax2, Sox2, and Wnt1), and the number of neural stem cells (NSCs) derived from B2h-oeM6A cells compared to control vector transfected human ES cells (B2h-Mock1). Our results show an increase in the number of differentiated neuronal cells (cholinergic, catecholaminergic, and GABAergic neurons) from NSCs derived from B2h-oeM6A cells. On the other hand, suppression of human GPM6A expression using a short hairpin RNA (shRNA) in human ES cells led to a decrease in both the expression of neuroectodermal-associated genes and the number of NSCs derived from B2h-shM6A cells. In addition, our results show a decrease in the number of differentiated neuronal cells from NSCs in B2h-shM6A cells compared to control vector transfected human ES cells (B2h-shNSP1). Moreover, overexpression or suppression of human GPM6A in human ES cells led to an increase or decrease, respectively, of neuronal migration. Hence, our findings suggest that expression level of GPM6A is, directly or indirectly, associated with the differentiation and neuronal migration of neurons derived from undifferentiated human ES cells. PMID- 19298181 TI - The structural and functional diversity of metabolite-binding riboswitches. AB - The cellular concentrations of certain metabolites are assiduously monitored to achieve appropriate levels of gene expression. Although proteins have long been known to act as sensors in this capacity, metabolite-binding RNAs, or riboswitches, also play an important role. More than 20 distinct classes of riboswitches have been identified to date, and insights to the molecular recognition strategies of a significant subset of these have been provided by detailed structural studies. This diverse set of metabolite-sensing RNAs is found to exploit a variety of distinct mechanisms to regulate genes that are fundamental to metabolism. PMID- 19298182 TI - Motors, switches, and contacts in the replisome. AB - Replisomes are the protein assemblies that replicate DNA. They function as molecular motors to catalyze template-mediated polymerization of nucleotides, unwinding of DNA, the synthesis of RNA primers, and the assembly of proteins on DNA. The replisome of bacteriophage T7 contains a minimum of proteins, thus facilitating its study. This review describes the molecular motors and coordination of their activities, with emphasis on the T7 replisome. Nucleotide selection, movement of the polymerase, binding of the processivity factor, unwinding of DNA, and RNA primer synthesis all require conformational changes and protein contacts. Lagging-strand synthesis is mediated via a replication loop whose formation and resolution is dictated by switches to yield Okazaki fragments of discrete size. Both strands are synthesized at identical rates, controlled by a molecular brake that halts leading-strand synthesis during primer synthesis. The helicase serves as a reservoir for polymerases that can initiate DNA synthesis at the replication fork. We comment on the differences in other systems where applicable. PMID- 19298183 TI - Biological and chemical approaches to diseases of proteostasis deficiency. AB - Many diseases appear to be caused by the misregulation of protein maintenance. Such diseases of protein homeostasis, or "proteostasis," include loss-of-function diseases (cystic fibrosis) and gain-of-toxic-function diseases (Alzheimer's, Parkinson's, and Huntington's disease). Proteostasis is maintained by the proteostasis network, which comprises pathways that control protein synthesis, folding, trafficking, aggregation, disaggregation, and degradation. The decreased ability of the proteostasis network to cope with inherited misfolding-prone proteins, aging, and/or metabolic/environmental stress appears to trigger or exacerbate proteostasis diseases. Herein, we review recent evidence supporting the principle that proteostasis is influenced both by an adjustable proteostasis network capacity and protein folding energetics, which together determine the balance between folding efficiency, misfolding, protein degradation, and aggregation. We review how small molecules can enhance proteostasis by binding to and stabilizing specific proteins (pharmacologic chaperones) or by increasing the proteostasis network capacity (proteostasis regulators). We propose that such therapeutic strategies, including combination therapies, represent a new approach for treating a range of diverse human maladies. PMID- 19298185 TI - Reproductive character displacement in genital morphology in Satsuma land snails. AB - When species diverge in allopatry and come into secondary contact, natural selection might favor character divergence to facilitate partner discrimination and avoid mating interference. This process is called reinforcement, which typically results in greater differentiation in mating-associated traits in areas of sympatry (reproductive character displacement). Such patterns are usually manifest in premating signals, but little is known about how interspecific mating affects the morphological evolution of genitalia. Here, we present evidence of reproductive character displacement in genital morphology in two land snail species of the genus Satsuma. Morphometric analyses clearly revealed greater sympatric divergences in penis and vagina lengths, regardless of the environmental, genetic, and geographic effects. This is the first demonstration of reproductive character displacement in genital characters in an organism other than insects and highlights the importance of analyzing previously overlooked genital morphologies in understanding the general mechanisms of prezygotic isolation in land snails. PMID- 19298189 TI - Cocoa flavanols and procyanidins can modulate the lipopolysaccharide activation of polymorphonuclear cells in vitro. AB - Flavanols and procyanidins isolated from cocoa have been reported to possess multiple activities potentially relevant to oxidant defenses, vascular function, and immune function. In a combination of in vivo and in vitro studies, we and others have observed that cocoa can be an anti-inflammatory modulator and that compounds in cocoa are capable of modulating eicosanoid production, platelet aggregation, and the pool size of nitric oxide. The present study extends these findings by examining the in vitro effects of cocoa procyanidins on polymorphonuclear cells (PMNs). PMNs, part of the innate arm of the immune system, represent 50-60% of the total peripheral white blood cells and are the first cells to be recruited to the sites of inflammation or injury secondary to bacterial infections. Herein, we demonstrate that certain flavanols and procyanidins isolated from cocoa can moderate a subset of signaling pathways derived from lipopolysaccharide (LPS) stimulation of PMNs, mainly, PMN oxidative bursts and activation markers, and they can influence select apoptosis mechanisms. We hypothesize that flavanols and procyanidins can decrease the impact of LPS on the N-formyl-Met-Leu-Phe-primed PMN ability to generate reactive oxygen species by partially interfering in activation of the mitogen-activated protein kinase pathway. PMID- 19298190 TI - Induction of cell cycle arrest in prostate cancer cells by the dietary compound isoliquiritigenin. AB - Isoliquiritigenin (ISL), a flavonoid chalcone that is present in licorice, shallot, and bean sprouts, is known to have antitumorigenic activities. The present study examined whether ISL alters prostate cancer cell cycle progression. DU145 human and MatLyLu (MLL) rat prostate cancer cells were cultured with various concentrations of ISL. In both DU145 and MLL cells treated with ISL, the percentage of cells in the G1 phase increased, and the incorporation of [(3)H]thymidine decreased. ISL decreased the protein levels of cyclin D1, cyclin E, and cyclin-dependent kinase (CDK) 4, whereas cyclin A and CDK2 expressions were unaltered in cells treated with ISL. The expression of the CDK inhibitor p27(KIP1) was increased in cells treated with 20 micromol/L ISL. In addition, treatment of cells with 20 micromol/L ISL for 24 hours led to G2/M cell cycle arrest. Cell division control (CDC) 2 protein levels remained unchanged. The protein levels of phospho-CDC2 (Tyr15) and cyclin B1 were increased, and the CDC25C level was decreased by ISL dose-dependently. We demonstrate that ISL promotes cell cycle arrest in DU145 and MLL cells, thereby providing insights into the mechanisms underlying its antitumorigenic activities. PMID- 19298191 TI - Effects of dietary Spirulina on vascular reactivity. AB - There are several reports suggesting that Spirulina (Arthrospira) may have a beneficial effect in the prevention of cardiovascular diseases. Here we review the results of studies on the effects of dietary Spirulina on the vasomotor reactivity of aortic rings excised from either lean or obese Wistar rats. We also review preliminary results on the effects of Spirulina intake on plasma lipids and blood pressure in humans. The results of the former studies strongly suggest that Spirulina induces a tone-related increase in the synthesis/release of nitric oxide by the endothelium as well as an increase in the synthesis/release of a vasodilating cyclooxygenase-dependent metabolite of arachidonic acid and/or a decrease in the synthesis/release of a vasoconstricting eicosanoid by the endothelium. In humans, Spirulina maxima intake decreases blood pressure and plasma lipid concentrations, especially triacylglycerols and low-density lipoprotein-cholesterol, and indirectly modifies the total cholesterol and high density lipoprotein-cholesterol values. PMID- 19298192 TI - Vascular reactivity is affected by dietary consumption of wild blueberries in the Sprague-Dawley rat. AB - We have previously reported that consumption of blueberry-enriched (BB) diets attenuates the arterial contractile response to alpha(1)-adrenergic stimuli and affects vasomotor tone via endothelium-related pathways. The present study was designed to evaluate vascular function and responsiveness in aortas of weanling male Sprague-Dawley rats fed a control (C) or a BB diet for 7 weeks. Vascular ring studies were conducted in 3-mm isolated rat aortic ring preparations to investigate vasoconstriction induced by L-phenylephrine (Phe) (10(-8)-3 x 10(-6) M) and vasorelaxation induced by acetylcholine (ACh) (10(-8)-3 x 10(-6) M). Agonists were used alone and in the presence of nitric oxide (NO) synthase and cyclooxygenase (COX) inhibitors. We observed a significantly diminished vasoconstrictor response to Phe in aortic rings from rats fed the BB diet. Inhibition of NO synthase but not COX caused a significant increase in the constrictor response in both dietary groups, with the BB group having the greater response. Similarly, the participation of the NO pathway in endothelium-dependent vasorelaxation induced by ACh was greater in the rats fed a BB diet, while COX inhibition showed no effect on maximum ACh-induced vasorelaxation in any diet group. The vessel sensitivity of BB aortic rings to the vasoconstrictor and vasodilator was significantly reduced when compared to controls. We have concluded that diets enriched with blueberries, fed for 7 weeks in Sprague-Dawley rats, seem to affect NO metabolic pathways in the aorta at basal and stimulated levels. PMID- 19298193 TI - Moderate coffee consumption lowers the likelihood of developing left ventricular systolic dysfunction in post-acute coronary syndrome normotensive patients. AB - The aim of the present work was to evaluate the association between coffee consumption and the development of left ventricular systolic dysfunction (LVSD) in patients who had had an acute coronary syndrome. During 2006-2007, 144 male (65 +/- 14 years) and 50 female (71 +/- 12 years) post-acute coronary syndrome patients who developed LVSD (ejection fraction <40%) after the cardiac event and 129 male (64 +/- 12 years) and 51 female (67 +/- 10 years) post-acute coronary syndrome patients without LVSD (ejection fraction >50%) were included in the study. Participants were consequently selected. Detailed information regarding their medical records, sociodemographic and anthropometric data, and various psychological and lifestyle characteristics (physical activity, smoking habits, etc.) were recorded. In particular, nutritional habits, including coffee consumption, were evaluated using a semiquantitative food-frequency questionnaire. Multi-adjusted analysis revealed that in normotensive patients coffee consumption of 1-2 cups/day was associated with 88% (95% confidence interval, 0.02-0.84) lower likelihood of developing LVSD and consumption of >3 cups/day with 90% (95% confidence interval, 0.01-0.88) lower likelihood for LVSD, compared with no history of consumption of coffee and after adjusting for various confounders. In contrast, in hypertensive patients coffee consumption of >3 cups/day was associated with 4.5-fold higher likelihood for developing LVSD (95% confidence interval, 0.89-22.58) as compared with no history of coffee consumption. Coffee consumption has opposite effects on the likelihood of developing LVSD in post-acute coronary syndrome patients depending on their blood pressure levels. PMID- 19298194 TI - Bovine colostrum prevents bacterial translocation in an intestinal ischemia/reperfusion-injured rat model. AB - This study evaluated whether or not bovine colostrum (BC) is able to treat or prevent intestinal barrier damage, bacterial translocation, and the related systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) in an intestinal ischemia/reperfusion (I/R)-injured rat model. Fifty Sprague-Dawley rats were used. The rats' intestinal I/R injuries were induced by clamping the superior mesenteric artery for 30 minutes. After 3 hours of reperfusion and then twice daily reclamping during the experiment, the experimental group was given BC (4 mL/kg/day) perorally, and the other groups received 0.9% saline and low fat milk (LFM) after intestinal I/R injury. Seventy two hours later we assessed (1) intestinal damage and intestinal permeability, (2) enteric bacterial count and bacterial translocation, (3) serum albumin, protein, and hepatic enzyme levels, (4) pathologic findings of ileum and lung, (5) activity of oxygen-free radical species, and (6) pro-inflammatory cytokines (tumor necrosis factor-alpha and interleukin-1beta). Intestinal damage, intestinal permeability, and bacterial translocation to other organs were significantly reduced in rats fed with BC after I/R when compared to rats fed LFM/saline after I/R (P < .05). In the evaluation of acute lung injury, neutrophils were found only in the lungs of the saline-fed group after I/R, and the wet/dry ratio of the lung tissue was significantly reduced in the BC-fed group after I/R compared to other I/R groups. A marked difference was found between LFM/saline-fed groups and BC-fed groups regarding malondialdehyde (P < .05) and pro-inflammatory cytokines (P < .01). In conclusion, BC may have beneficial effects in treating and preventing intestinal barrier damage, bacterial translocation and the related SIRS and MODS in the intestinal I/R injured rat model. PMID- 19298195 TI - Moringa oleifera leaf extract prevents isoproterenol-induced myocardial damage in rats: evidence for an antioxidant, antiperoxidative, and cardioprotective intervention. AB - The present study evaluated cardioprotective effect of lyophilized hydroalcoholic extract of Moringa oleifera in the isoproterenol (ISP)-induced model of myocardial infarction. Wistar albino male rats were divided into three groups and orally fed saline once daily alone (sham) or with ISP (ISP control) or ISP with M. oleifera (200 mg/kg), respectively, for 1 month. On days 29 and 30 of administration, rats of the ISP control and M. oleifera-ISP groups were administered ISP (85 mg/kg, s.c.) at an interval of 24 hours. On day 31, hemodynamic parameters (mean arterial pressure [MAP], heart rate [HR], left ventricular end-diastolic pressure [LVEDP], and left ventricular peak positive [(+) LV dP/dt] and negative [(-) LV dP/dt] pressures were recorded. At the end of the experiment, the animals were sacrificed, and hearts were excised and processed for biochemical, histopathological, and ultrastructural studies. Chronic treatment with M. oleifera demonstrated mitigating effects on ISP-induced hemodynamic [HR, (+) LV dP/dt, (-) LV dP/dt, and LVEDP] perturbations. Chronic M. oleifera treatment resulted in significant favorable modulation of the biochemical enzymes (superoxide dismutase, catalase, glutathione peroxidase, lactate dehydrogenase, and creatine kinase-MB) but failed to demonstrate any significant effect on reduced glutathione compared to the ISP control group. Moringa treatment significantly prevented the rise in lipid peroxidation in myocardial tissue. Furthermore, M. oleifera also prevented the deleterious histopathological and ultrastructural perturbations caused by ISP. Based on the results of the present study, it can be concluded that M. oleifera extract possesses significant cardioprotective effect, which may be attributed to its antioxidant, antiperoxidative, and myocardial preservative properties. PMID- 19298196 TI - Effects of trans-10,cis-12 conjugated linoleic acid on body composition in genetically obese mice. AB - Conjugated linoleic acid (CLA) has shown a number of biologically beneficial effects, including prevention of obesity. The purpose of this study was to test effects of dietary supplementation of 0.5% trans-10,cis-12 CLA in a high fat diet in neuronal basic helix-loop-helix 2 knock-out animals (N2KO), which is a unique animal model representing adult-onset inactivity-related obesity. Eight wild-type (WT) and eight N2KO female mice were fed either 0.5% trans-10,cis-12 CLA containing diet or control diet (with 20% soybean oil diet) for 12 weeks. Body weights, food intake, adipose tissue weights, body compositions, and blood parameters were analyzed. Overall, N2KO animals had greater body weights, food intake, adipose tissue weights, and body fat compared to WT animals. CLA supplementation decreased overall body weights and total fat, and the effect of dietary CLA on adipose tissue reduction was greater in N2KO than in WT mice. Serum leptin and triglyceride levels were reduced by CLA in both N2KO and WT animals compared to control animals, while there was no effect by CLA on serum cholesterol. The effect of CLA to lower fat mass, increase lean body mass, and lower serum leptin and triglycerides in sedentary mice supports the possibility of using CLA to prevent or alleviate ailments associated with obesity. PMID- 19298197 TI - Effects of soy phytoestrogens on reference memory and neuronal cholinergic enzymes in ovariectomized rats. AB - The effects of soy phytoestrogens on Morris water maze (MWM) performance and neuronal cholinergic enzyme activities and immunoreactivity were studied in ovariectomized (OVX) rats. The rats were assigned to four groups fed control diet (CD), 3.9 mg/kg 17beta-estradiol diet (E2), 263.4 mg/kg soy phytoestrogens diet (SP1), and 526.9 mg/kg soy phytoestrogens diet (SP2). In the MWM task, escape latency and path length were significantly less in the E2 and SP2 groups than in the CD group on the second day. Choline acetyltransferase (ChAT) activity in the cerebral cortex and ChAT immunoreactivity in the diagonal band of Broca were significantly greater in the E2, SP1, and SP2 groups than in the CD group. Acetylcholinesterase activity in the hippocampus in the E2, SP1, and SP2 groups was significantly lower than in the CD group. This study suggests that soy phytoestrogens affect the reference memory and neuronal cholinergic system in OVX rats. PMID- 19298198 TI - Panax ginseng protects against global ischemia injury in rat hippocampus. AB - Based on the use of Panax ginseng C.A. Meyer (Family Araliaceae) for the treatment of stroke in traditional Korean medicine, the present study was carried out to evaluate neuroprotective effects of P. ginseng after transient global cerebral ischemia using the four-vessel occlusion rat model. Nissl staining, lipid peroxidation (malondialdehyde [MDA] formation), and activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) of rat brain were assessed. Ethanolic P. ginseng extract (200 mg/kg, i.p.) significantly protected CA1 neurons against 10 minutes of transient forebrain ischemia as demonstrated by measuring the density of neuronal cells. P. ginseng also significantly decreased the level of MDA and increased the expression of GPx and SOD. These results suggest that P. ginseng might be neuroprotective against cerebral ischemia induced injury in rat brain by decreasing lipid peroxides and increasing the expression of GPx and SOD. PMID- 19298199 TI - Antioxidant and inhibitory effects of aqueous extracts of Salvia officinalis leaves on pro-oxidant-induced lipid peroxidation in brain and liver in vitro. AB - The present study sought to determine the antioxidant activity and protective ability of water-extractable phytochemicals from Salvia officinalis leaves (strongly aromatic leaves used in flavoring cooked foods) on lipid peroxidation induced by some pro-oxidants in rat brain and liver homogenates in vitro. Aqueous extracts of the leaves were prepared, and the ability of the extract to inhibit 25 microM FeSO(4)- and 7 microM sodium nitroprusside-induced lipid peroxidation in isolated rat brain and liver was determined. Subsequently, the ascorbic acid content, total phenol content, reducing power, Fe(II) chelating, and .OH radical scavenging ability of the extracts were determined as indices of antioxidant activity. The results of the study revealed that the extract inhibited malondialdehyde (MDA) production in basal and pro-oxidant-induced lipid peroxidation in the brain and liver in a dose-dependent manner. The percentage induction of lipid peroxidation by Fe(II) and sodium nitroprusside was higher in the brain than the liver; however, the level of inhibition of MDA production in the brain was significantly (P < .05) higher than the liver. The ascorbic acid (10.3 +/- 2.5 mg/g) and total phenol (7.6 +/- 1.2 mg/g) contents of the leaves were high; likewise, the aqueous extract had high reducing power and Fe(II) chelating ability but low .OH radical scavenging ability. This antioxidant and protective effect of this leaf could be harnessed in the management and prevention of degenerative diseases associated with oxidative stress. PMID- 19298200 TI - Cuscuta chinensis extract promotes osteoblast differentiation and mineralization in human osteoblast-like MG-63 cells. AB - The aim of the present study was to investigate whether the aqueous extract of To Sa-Za (TSZ-AE), the seed of Cuscuta chinensis Lam., which is a traditional medicinal herb commonly used in Korea and other oriental countries, could induce osteogenic activity in human osteoblast-like MG-63 cells. TSZ-AE treatment mildly promoted the proliferation of MG-63 cells at doses of 500 and 1,000 microg/mL in the 24-hour culture period. Dose-dependent increases in alkaline phosphatase (ALP) activity and collagen synthesis were shown at 48 and 72 hours of incubation. The release of bone morphogenetic protein (BMP)-2 but not osteocalcin in the MG-63 cells was induced by TSZ-AE at 72 hours (100-1,000 microg/mL). In addition, TSZ-AE markedly increased mRNA expression of ALP, collagen, and BMP-2 in the MG-63 cells in a dose-dependent manner. Mineralization in the culture of MG-63 cells was significantly induced at 500 and 1,000 microg/mL TSZ-AE treatment. In conclusion, this study shows that TSZ-AE enhanced ALP activity, collagen synthesis, BMP-2 expression, and mineralization in MG-63 cells. These results strongly suggest that C. chinensis can play an important role in osteoblastic bone formation and may possibly lead to the development of bone forming drugs. PMID- 19298201 TI - Ethanol extract of Allium sativum attenuates testicular and liver toxicity induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in rats. AB - The present study was carried out to investigate the protective role of garlic (Allium sativum) ethanol extract (GE) in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced hepatic and testicular toxicity. A total of 60 male rats (Sprague Dawley, weighing 200 +/- 10 g) were divided into six equal groups. The normal control group (NC) received vehicle (intraperitoneally) and saline (perorally). A predetermined dosage of TCDD (40 microg/kg of body weight, i.p.) was administered to single TCDD-treated (TT) and test (GE) groups. GE was administered (perorally) at daily doses of 5 (GE5), 10 (GE10), 20 (GE 20), or 40 (GE40) mg/kg of body weight for 5 weeks, starting 1 week before the TCDD exposure. Decreases in body weight gain (P < .01) and testicular weight (P < .01) induced by TCDD were greatly attenuated by GE (P < .05-.01). TCDD-induced decreases in spermatogenesis related panels--Johnsen's score, seminiferous tubular size, ratio of tubules with sperm, and sperm count/tubule--were greatly improved by GE treatment in a dose dependent manner in the rats. TCDD-induced increases in serum cholesterol and triglyceride levels and glutamic oxaloacetate activity were also suppressed by GE (P < .05-.01). These results indicate that administration of garlic to TCDD exposed rats attenuates testicular and hepatic damage, suggesting that garlic might be a useful agent that can protect human health from toxic responses induced by environmental pollutants. PMID- 19298202 TI - Legume-, fish-, or high-protein-based hypocaloric diets: effects on weight loss and mitochondrial oxidation in obese men. AB - The nutritional composition of dietary intake could produce specific effects on metabolic variables such as mitochondrial oxidation, whose understanding could contribute to apply more individualized weight-lowering strategies. This study assessed the effects of four hypocaloric diets with high protein content or different food distribution on metabolic changes and mitochondrial oxidation accompanying weight loss. Thirty-five obese men (body mass index of 31.8 +/- 3.0 kg/m(2) and 38 +/- 7 years old) were randomly assigned to one of the four treatments (8 weeks): control diet (C-diet); legume diet (L-diet); fatty fish diet (FF-diet); or high-protein diet (HP-diet). Body composition, blood pressure, resting energy expenditure, mitochondrial oxidation, blood biomarkers, and dietary intake were assessed. The HP-diet and L-diet achieved the greater body weight reduction (-8.4 +/- 1.2% and -8.3 +/- 2.9%, respectively), as compared to the C-diet (-5.5 +/- 2.5%; P = .042). The high-density lipoprotein cholesterol concentrations were reduced in all dietary groups except for the FF-diet. Total and low-density lipoprotein cholesterol levels were significantly improved by the L-diet (P < .05), while the homeostatic model assessment index of insulin resistance value was significantly reduced in those men following the HP-diet. Mitochondrial oxidation was specifically activated by the HP-diet and L-diet at the end of the study. Interestingly, a lineal regression model explained about 25% (P = .029) of the mitochondrial oxidation variability as influenced by the diet changes once adjusted by resting energy expenditure. The specific consumption of legumes or high protein content within a hypocaloric diet could activate mitochondrial oxidation, which could involve additional benefits to those associated with the weight reduction. PMID- 19298203 TI - Antihyperglycemic effects of fruits of privet (Ligustrum obtusifolium) in streptozotocin-induced diabetic rats fed a high fat diet. AB - The protective effects of freeze-dried privet (Ligustrum obtusifolium) fruits (PFs) were observed in streptozotocin (STZ)-induced diabetic rats on a high fat diet by measuring levels of blood glucose, serum insulin, fructosamine, and hepatic reactive oxygen species generating and scavenging enzyme activities. A PF supplemented diet was prepared by mixing an AIN-76 diet with powdered PF (final concentration, 1% or 2%). It was fed to STZ-induced diabetic rats on a high fat diet for 6 weeks. Diabetic animals receiving the PF-supplemented diet showed a significant increase in body weight, feed efficiency ratio, liver, kidney, and heart weight, and serum glucose, insulin, and fructosamine levels compared with high fat diet-fed diabetic animals. The treatment with PF showed improved hepatic glutathione S-transferase, superoxide dismutase, and xanthine oxidase activities as well as glutathione and lipid peroxide levels in the diabetic animals. Intracellular swelling and vacuole formation in diabetic pancreatic beta- and delta-cells were ameliorated by the PF-supplemented diet. Furthermore, necrosis of tubular epithelial cells and dilatation of luminal space in diabetic kidneys exhibited near-noninjured condition. This is the first time an antihyperglycemic effect of L. obtusifolium fruit in STZ-induced diabetic rats has been identified. PMID- 19298204 TI - Zinc-deficient diet decreases fetal long bone growth through decreased bone matrix formation in mice. AB - This study evaluated the effects of zinc on skeletal development during fetal development in pregnant ICR mice fed a zinc-deficient (3 mg/kg) or zinc-adequate (30 mg/kg) diet. We also included a group pair-fed with the zinc-deficient group to control for decreased appetite due to zinc deficiency. Developing fetuses at embryonic day 18.5 were removed by cesarean section, and the skeletal development was evaluated by histological analysis as well as by body weight and longitudinal growth measurement. Reduced maternal food intake in the zinc-deficient and pair fed groups resulted in a marked and significant (P < .05) decrease in fetal weight compared to that of the zinc-adequate group. However, fetal length retardation in the pair-fed group was less marked than in the zinc-deficient group, suggesting that reduced supply of zinc from maternal circulation may play a role in longitudinal growth through skeletal development. The fetal developing tibia of the zinc-deficient group showed marked shortening of diaphysis and a mild narrowing of the hypertrophic chondrocyte zone width with increased osteoclast number, but there was no influence on the mineralization of bone matrix. This may be the result of reduced activation of osteoblasts and maturation of chondrocytes with increased osteoclastic activity, suggesting that zinc deficiency during the fetal development has a greater impact on the matrix formation of bone than the mineralization of bone matrix. PMID- 19298205 TI - Evaluation of the peroxynitrite scavenging activity of heat-processed ginseng. AB - To ascertain the principal active peroxynitrite (ONOO(-)) scavenging components of heat-processed Panax ginseng C.A. Meyer (sun ginseng [SG]), the ONOO(-) scavenging activities of fractions and components of SG were compared. The results demonstrated that the ONOO(-) scavenging ability of SG was due to its ether fraction containing phenolic compounds. High-performance liquid chromatography analysis and ONOO(-) scavenging activity tests of the phenolic acids contained in SG identified vanillic acid, ferulic acid, p-coumaric acid, syringic acid, and maltol as the main active ONOO(-) scavenging components of SG. The ONOO(-) scavenging activities of phenolic acids and maltol were dependent on the degrees of their proton donating ability. PMID- 19298206 TI - Antiplasmodial and antidiabetic activities of ethanolic leaf extract of Heinsia crinata. AB - The ethanolic leaf extract of Heinsia crinata, grown particularly for the leaf in Niger Delta region of Nigeria, was evaluated for antiplasmodial activity in Plasmodium berghei-infected mice as well as for hypoglycemic and antidiabetic activities in alloxan-induced diabetic rats. H. crinata (450-1,350 mg/kg/day) exhibited significant (P < .05) blood schizonticidal activity in both the 4-day early infection test and established infection with a considerable mean survival time, though not comparable to that of the standard drug, chloroquine (5 mg/kg/day). Treatment of alloxan-induced diabetic rats with the leaf extract caused a significant (P < .01) reduction in fasting blood glucose levels of normal and alloxan-induced diabetic rats both in an acute study and with prolonged treatment (2 weeks). The activity of the extract was comparable to that of the reference drug, glibenclamide, during the acute study but was not comparable to that of the standard drug during prolonged treatment. The results suggest that the leaf extract of H. crinata possesses significant (P < .05) antiplasmodial as well as hypoglycemic and antidiabetic effects that can be employed in health care. PMID- 19298207 TI - Methionine-induced hyperhomocysteinemia modulates lipoprotein profile and oxidative stress but not progression of atherosclerosis in aged apolipoprotein E knockout mice. AB - It is documented that hyperhomocysteinemia (HHcy) is an independent risk factor for atherosclerosis, but whether elevated plasma homocysteine contributes to the progression of atherosclerosis in aged animals with hypercholesterolemia is still unknown. HHcy was induced in apolipoprotein E (ApoE) knockout mice (male, 32 weeks old) by feeding 2% methionine/low folate (1 mg/kg) diet for 20 weeks. HHcy induced by methionine feeding significantly increased oxidative stress, as measured by thiobarbituric-reactive substances in livers (P < .05) and genetic expression of Cu,Zn-superoxide dismutase, in methionine-fed animals compared with controls (P < .05). Furthermore, lipoprotein profiles were changed, in that low density lipoprotein-cholesterol was shifted to very low-density lipoprotein in the methionine-supplemented group. However, nuclear factor kappaB activity, atherosclerotic lesions, hepatic glutathione level, lipid profiles, and activities of aspartate aminotransferase and alanine aminotransferase were not significantly different. These findings suggest that HHcy induced by methionine may promote disturbances in lipid peroxidation and modify lipoprotein metabolism but not contribute to the progression of atherosclerotic lesion in aged ApoE knockout mice. PMID- 19298208 TI - Oxidative and antioxidant status in plasma of runners: effect of oral supplementation with natural antioxidants. AB - Aerobic exercise increases free radical production as a consequence of enhanced oxygen consumption. If free radical formation exceeds antioxidant capacity, lipids, proteins, and DNA may be oxidized. Oxidative stress is widely recognized as a factor in many degenerative human diseases. The role of dietary antioxidants in protection against disease is a topic of continuing interest. In fact, there is epidemiological evidence correlating a higher intake of nutrients possessing antioxidant abilities with a lower incidence of various human diseases. This study was directed at investigating whether changes in plasma antioxidant capacity and oxidative stress markers occur in voluntary wheel runners, before and after oral supplementation with lycopene and isoflavones. For this purpose, plasma antioxidant capacity and oxidative stress markers were assessed in long distance runners at the end of a 60-minute run. Comparisons were made between runners before and after 60 days of supplementation with lycopene and isoflavones. DNA damage in blood cells of the same samples was also evaluated by comet assay. This investigation shows that oral supplementation with lycopene and soy-derived isoflavones significantly reduced lipid peroxidation and enhanced plasma nonproteic antioxidant defense. PMID- 19298209 TI - Cytoprotective and antioxidant activity of seabuckthorn (Hippophae rhamnoides L.) flavones against tert-butyl hydroperoxide-induced cytotoxicity in lymphocytes. AB - This study was designed to determine the cytoprotective activity of flavones of seabuckthorn (Hippophae rhamnoides L.) against tert-butyl hydroperoxide (tert BOOH), used as an oxidant to induce oxidative damage, with lymphocytes as the model system. Addition of tert-BOOH (250 microM) to the cells resulted in enhanced cytotoxicity and free radical production. The intracellular calcium levels, caspase activity, and apoptosis were significantly increased following tert-BOOH treatment. Seabuckthorn flavones at the concentration of 100 microg/mL significantly inhibited tert-BOOH-induced cytotoxicity and free radical production and also restored the antioxidant status to that of control cells. Seabuckthorn flavones also significantly restricted tert-BOOH-induced apoptosis by decreasing intracellular calcium levels and caspase activity. The extract also decreased tert-BOOH-induced formation of DNA breaks by 30%. These observations suggest that the flavones of seabuckthorn have marked cytoprotective properties, which could be attributed to the antioxidant activity. PMID- 19298210 TI - Protective effects against H2O2-induced damage by enzymatic hydrolysates of an edible brown seaweed, sea tangle (Laminaria japonica). AB - Enzymatic hydrolysates of Laminaria japonica were evaluated for antioxidative activities using hydroxyl radical scavenging activity and protective effects against H(2)O(2)-induced DNA and cell damage. In addition, activities of antioxidative enzymes, including catalase, glutathione peroxidase, and glutathione S-transferase, of the enzymatic hydrolysates from L. japonica were also estimated. L. japonica was first enzymatically hydrolyzed by seven carbohydrases (Dextrozyme, AMG, Promozyme, Maltogenase, Termamyl, Viscozyme, and Celluclast [all from Novo Co., Novozyme Nordisk, Bagsvaerd, Denmark]) and five proteinases (Flavourzyme, Neutrase, Protamex, Alcalase [all from Novo Co.], and pancreatic trypsin). The hydroxyl radical scavenging activities of Promozyme and pancreatic trypsin hydrolysates from L. japonica were the highest as compared to those of the other carbohydrases and proteinases, and their 50% inhibitory concentration values were 1.67 and 317.49 mug/mL, respectively. The pancreatic trypsin hydrolysates of L. japonica exerted a protective effect on H(2)O(2) induced DNA damage. We also evaluated the protective effect on hydroxyl radical induced oxidative damage in PC12 cells via propidium iodide staining using a flow cytometer. The AMG and pancreatic trypsin hydrolysates of L. japonica dose dependently protected PC12 cells against cell death caused by hydroxyl radical induced oxidative damage. Additionally, we analyzed the activity of antioxidative enzymes such as catalase, glutathione peroxidase, and the phase II biotransformation enzyme glutathione S-transferase in L. japonica-treated cells. The activity of all antioxidative enzymes was higher in L. japonica-treated cells compared with the nontreated cells. These results indicate that enzymatic hydrolysates of L. japonica possess antioxidative activity. PMID- 19298211 TI - Antioxidant capacity of fresh and processed fruit bodies and mycelium of Auricularia auricula-judae (Fr.) Quel. AB - Auricularia auricula-judae is currently grown in Malaysia. In the present study, the methanolic extracts from fruit bodies (fresh, oven-dried, and freeze-dried) and mycelium of A. auricula-judae were evaluated for their antioxidant capacities based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity and ferric reducing antioxidant power (FRAP) assay. The total phenolic content in the extracts were also measured. The extract of freeze-dried fruit bodies of A. auricula-judae had potent DPPH free radical scavenging activity with a 50% effective concentration of 2.87 mg/mL, whereas the FRAP value of A. auricula judae mycelium was 5.22 micromol of FeSO(4).7H(2)O equivalents/g of mycelium sample. Further, a positive correlation (R(2) = 0.7668) between FRAP level of A. auricula-judae extracts and the total phenolic contents was observed. Thus the method of processing of fresh fruit bodies had an effect on the antioxidant potential of A. auricula-judae. PMID- 19298212 TI - Oxygen and nitrogen reactive species are effectively scavenged by Eucalyptus globulus leaf water extract. AB - Eucalyptus globulus Labill. (Family Myrtaceae) is a plant of Australian origin, with a reported therapeutic use in airway inflammatory diseases. Considering that reactive oxygen species (ROS) and reactive nitrogen species (RNS) have been implicated in the pathogenesis of airway inflammatory diseases such as asthma and chronic obstructive pulmonary disease, an effective scavenging activity against these reactive species may contribute for the therapeutic effect of this plant. In the present study, a water extract of E. globulus leaves was evaluated for its putative in vitro scavenging effects on ROS (HO(*), O(2)(*-), ROO(*), and H(2)O(2)) and RNS ((*)NO and ONOO(-)) and on 2,2-diphenyl-1-picrylhydrazyl radical (DPPH). Qualitative and quantitative analyses of the extract's phenolic composition were also performed. The Eucalyptus leaf water extract presented a remarkable capacity to scavenge all the reactive species tested, with all the 50% inhibitory concentrations being found at the mug/mL level. Phytochemical analysis showed the presence of polyphenols such as flavonoids (rutin and quercitrin) and phenolic acids (chlorogenic acid and ellagic acid), which may be partially responsible for the observed antioxidant activity. These observations provide further support, beyond the well-known antibacterial and antiviral activities of the Eucalyptus plant, for its reported use in traditional medicine such as in the treatment of airway inflammatory diseases, considering the important role of ROS and RNS in the inflammatory process, although further studies are needed to prove the bioavailability of the antioxidants/antibacterial compounds of the extract as well as the ability of the active compounds to reach specific tissues and to act in them. PMID- 19298213 TI - Potentials of Kerstingiella geocarpa as a health food. AB - The seed of Kerstingiella geocarpa was analyzed to determine its proximate and amino acid compositions. The seed was found to be a high source of crude protein (21.3%) and very low in crude fat (1.0%). Arginine, an amino acid for pediatric growth, was the most concentrated amino acid in the seed. Arginine (9.3 g/100 g of crude protein), histidine (2.1 g/100 g of crude protein), and phenylalanine (3.2 g/100 g of crude protein) concentrations in the seed were found in higher levels than the recommended Food and Agriculture Organization/World Health Organization amino acid requirement pattern for the preschool age group. The total essential amino acid content in K. geocarpa seed was 42.0%. The lysine:arginine ratio calculated to estimate the seed atherogenic potential was 0.3. This shows that K. geocarpa seed protein will not exert a hypercholesterolemia effect on the consumer. Futhermore, incorporation of the seed into diets of experimental rats resulted in hematological parameters comparable with those of casein control-fed rats. These findings show this seed could be an important source of food that holds promise in medicinal nutrition. PMID- 19298214 TI - Antioxidant activity and phenolic and mineral content of rose grape juice. AB - There are many studies related to the antioxidant activity of grape products; however, they concern only purple and white grape varieties. Up to now, there are no reports of studies on the Goethe rose grape variety, either on its antioxidant activity or on its phenolic and mineral quantification. Thus, the purpose of this study was to evaluate in vitro and in vivo antioxidant activity, as well as to quantify total phenolic compounds, ascorbic acid, and mineral content, in a Goethe rose grape juice. The results obtained showed that the Goethe rose grape juice is a great polyphenol source, which contains catechin, epicatechin, and procyanidins (B(1), B(2), B(3), and B(4)). Of all metals analyzed, potassium, calcium, magnesium, and iron showed the highest values. We found that this rose grape juice shows an important antioxidant activity in in vitro (2,2-diphenyl-1 picrylhydrazyl radical scavenging activity) and in vivo (using the Saccharomyces cerevisiae yeast cells) assays. The antioxidant activity could be explained by the significant phenolic content and ascorbic acid levels found in the juice. The results showed that rose grape juice is an excellent antioxidant source, which could contribute to the prevention of many diseases related to oxidative stress, such as atherosclerosis and Parkinson's disease. PMID- 19298215 TI - Effectiveness of Cymbopogon citratus L. essential oil to inhibit the growth of some filamentous fungi and yeasts. AB - Lemon grass (Cymbopogon citratus L.) oil has been known as having therapeutic and antibacterial properties, and its antifungal activity is currently the subject of renewed interest. This study aimed to verify the effectivenesses of C. citratus essential oil to inhibit the growth/survival of some fungi (Alternaria alternata, Aspergillus niger, Fusarium oxysporum, and Penicillium roquefortii) and yeasts (Candida albicans, Candida oleophila, Hansenula anomala, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Saccharomyces uvarum, and Metschnikowia fructicola). C. citratus essential oil showed effectiveness in inhibiting the growth of all fungi by disc diffusion and broth dilution bioassay. Minimum inhibitory and minimum fungicidal concentrations between 0.062 and 20 microL/mL were determined. The Clinical and Laboratory Standards Institute agar-based method was also applied for A. niger and C. albicans. Data show the strong antifungal properties of lemon grass oil (C. citratus) in vitro. PMID- 19298216 TI - Antioxidant activity, phenolic content, and peroxide value of essential oil and extracts of some medicinal and aromatic plants used as condiments and herbal teas in Turkey. AB - The antioxidant activity, total peroxide values, and total phenol contents of several medicinal and aromatic plant essential oil and extracts from Turkey were examined. Total phenolic contents were determined using a spectrophotometric technique and calculated as gallic acid equivalents. Total antioxidant activity of essential oil and extracts varied from 0.6853 to 1.3113 and 0.3189 to 0.6119 micromol of Trolox equivalents/g, respectively. The total phenolic content of essential oil ranged from 0.0871 to 0.5919 mg of gallic acid/g dry weight. However, the total phenolic contents of extracts were found to be higher compared with those of essential oils. The amount of total peroxide values of oils varied from 7.31 (pickling herb) to 58.23 (bitter fennel flower) mumol of H(2)O(2)/g. As a result, it is shown that medicinal plant derivatives such as extract and essential oils can be useful as a potential source of total phenol, peroxide, and antioxidant capacity for protection of processed foods. PMID- 19298217 TI - Constituents of leaves and flowers essential oils of Helichrysum pallasii (Spreng.) Ledeb. growing wild in Lebanon. AB - The chemical compositions of the essential oils obtained from leaves and flowers of Helichrysum pallasii were analyzed by gas chromatography and gas chromatography-mass spectrometry. Among the 102 identified constituents, hexadecanoic acid (16.2%), (Z,Z)-9,12-octadecadienoic acid (6.8%), tetradecanoic acid (2.6%), and (Z)-caryophyllene (4.2%) were the main constituent of the oil from leaves, while in the oil from flowers hexadecanoic acid (14.7%), (Z,Z)-9,12 octadecadienoic acid (14.2%), (Z)-caryophyllene (3.6%), and delta-cadinene (3.1%) predominated. The oils were both characterized by sesquiterpenes (33.4% for leaves and 33.7% for flowers, respectively) and fatty acids and esters (30.3% in leaves and 35% in flowers, respectively). The in vitro activity of the essential oils of the plant against some microorganisms in comparison with chloramphenicol by the broth dilution method was determined. The oils exhibited a weak activity as inhibitors of growth of Staphylococcus epidermidis in vitro (minimum inhibitory concentration = 100 microg/mL). PMID- 19298218 TI - Nitric oxide-releasing nonsteroidal anti-inflammatory drugs: gastrointestinal sparing potential drugs. AB - The nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most commonly prescribed drugs in the world. Their use as anti-inflammatory, antipyretic, antithrombotic, and analgesic agents is, however, restricted by their adverse effects on gastrointestinal (GI) tract. To overcome this problem with NSAIDs, two new strategies in the development of these drugs are explained. The first approach is the development of selective inhibitors of the enzyme cyclooxygenase (COX)-2, the inducible isoform of the prostaglandin G/H synthetase enzyme. COX exists in two isoforms, viz., COX-1 and COX-2. Selective inhibitors of COX-2 will suppress prostaglandin synthesis at the sites of inflammation, but they will not interfere with the activity of COX-1, in tissues like the GI tract. The use of selective COX-2 inhibitors, however, lacks the cardioprotective effects mediated by COX-1. An alternate approach to reduce the GI toxicity of NSAIDs is the development of nitric oxide (NO)-releasing NSAIDs. It has been reported that NO does not affect the gastroduodenal mucosa and produces the same effects as endogenous prostaglandins. The present article focuses on the NO-releasing NSAIDs, which are from the conventional class of COX-2 inhibitors that have potential clinical applications, as well as some drug candidates still under development. PMID- 19298219 TI - Economic and clinical evaluation of fondaparinux vs. enoxaparin for thromboprophylaxis following general surgery. AB - PURPOSE: Patients undergoing general surgical procedures are at increased risk for venous thromboembolism (VTE). Compliance rates with established guidelines for VTE thromboprophylaxis in patients at moderate-to-high risk are notably low. Recent literature has demonstrated that fondaparinux is associated with lower costs and fewer VTEs than enoxaparin in patients undergoing major orthopedic surgery (MOS), but data are limited in patients undergoing general surgery. This study was conducted to evaluate the cost implications and relative real-world effectiveness of fondaparinux vs. enoxaparin in general surgery patients. METHODS: Data were obtained from inpatient billing records from over 500 hospitals using Premier's Perspective Comparative Database. Patients hospitalized for general surgery between July 1, 2003 and January 31, 2006 were eligible for inclusion. Eligible patients were included if they received fondaparinux or enoxaparin after their general surgery date. Patients were excluded if they received both anticoagulants on their first day of therapy, were <18 years of age on the surgery date, or did not have data 6 months prior and 1 month post hospitalization. Included patients were stratified into two cohorts based on their first anticoagulant, fondaparinux or enoxaparin. Patients were matched in each group on 1:1 case-control matching based on propensity scores. RESULTS: A total of 5364 patients were included (n = 2682 for each cohort) from 326 unique hospitals. Average total costs per patient for the fondaparinux group were significantly lower than the enoxaparin group ($15 156 vs. 17 741, p < 0.0001). Patients receiving fondaparinux were significantly less likely to experience a VTE (2.80 vs. 3.77%, p = 0.046, a 35% relative risk reduction). No significant differences in bleeding events between the cohorts were observed (p = 0.6047), and no significant differences in all-cause inpatient death were noted (p = 0.3673). CONCLUSION: Fondaparinux was associated with significantly lower costs and fewer VTEs compared to enoxaparin without an increase in bleed rates or all cause inpatient mortality. The findings from this study are limited by the retrospective study design and should only be generalized to a similar patient population. PMID- 19298220 TI - N-terminal domains of CCN family 2/connective tissue growth factor bind to aggrecan. AB - CCN2/CTGF (CCN family 2/connective tissue growth factor) is a multi-cellular protein with a broad range of activities. It modulates many cellular functions, including proliferation, migration, adhesion and extracellular matrix production, and it is thus involved in many biological and pathological processes. In particular, CCN2/CTGF is essential for normal skeletal development. To identify CCN2/CTGF-interactive proteins capable of modulating its action in cartilage, we carried out a yeast two-hybrid screening using CCN2/CTGF peptide as a bait and a cDNA library from a chondrocytic cell line, HCS-2/8. In the present paper, we report the identification of aggrecan, which is a major proteoglycan of the extracellular matrix in cartilage, as a CCN2/CTGF-binding protein. Among the four domains of CCN2/CTGF, the IGFBP [IGF (insulin-like growth factor)-binding protein like] and/or VWC (von Willebrand factor type C) domains had a direct interaction with aggrecan in a yeast two-hybrid assay. The results of a solid-phase-binding assay using aggrecan-coated plates also showed binding to recombinant CCN2/CTGF in a dose-dependent manner. rIGFBP (recombinant IGFBP) and rVWC (recombinant VWC) module peptides had stronger binding to aggrecan compared with rTSP1 (recombinant thrombospondin type 1 repeat) and rCT (recombinant C-terminal cystine knot) module peptides. SPR (surface plasmon resonance) analysis showed the direct interaction between the CCN2/CTGF and aggrecan, and ectopically overexpressed CCN2/CTGF and AgG3 (G3 domain of aggrecan) confirmed their binding In vivo. Indirect immunofluorescence analysis indicated that CCN2/CTGF was extracellularly co-localized with aggrecan on HCS-2/8 cells. The rIGFBP-rVWC peptide effectively enhanced the production and release of aggrecan compared with the rTSP-rCT peptide in chondrocytes. These results indicate that CCN2/CTGF binds to aggrecan through its N-terminal IGFBP and VWC modules, and this binding may be related to the CCN2/CTGF-enhanced production and secretion of aggrecan by chondrocytes. PMID- 19298221 TI - The upregulation of receptor activator NF-kappaB ligand expression by interleukin 1alpha and Porphyromonas endodontalis in human osteoblastic cells. AB - AIM: To investigate the receptor activator of nuclear factor-kappa B (NF-kappaB) ligand (RANKL) in osteoblastic cells stimulated with inflammatory mediators. METHODOLOGY: The expression of RANKL in human osteoblastic cell line U2OS stimulated by pro-inflammatory cytokine interleukin (IL)-1alpha and black pigmented bacteria Porphyromonas endodontalis was investigated by Western blot and enzyme-linked immunosorbent assay (ELISA). The significance of the results obtained from control and treated groups was statistically analysed by the paired Student's t-test. RESULTS: IL-1alpha was found to upregulate RANKL production in U2OS cells (P < 0.05). Investigations of the time dependence of RANKL expression in IL-1alpha-treated cells revealed a rapid accumulation of RANKL protein after 1 h of exposure; it remained elevated throughout the 24-h incubation period shown by Western blot and ELISA. In addition, P. endodontalis also increased RANKL expression in U2OS cells after 4-h incubation period demonstrated by Western blot and ELISA (P < 0.05). CONCLUSIONS: IL-1alpha and P. endodontalis may be involved in developing apical periodontitis through the stimulation of RANKL production. PMID- 19298222 TI - Fresh-frozen, optimal cutting temperature (OCT) compound-embedded bone marrow aspirates: a reliable resource for morphological, immunohistochemical and molecular examinations. AB - The usefulness of fresh-frozen, optimal cutting temperature (OCT) compound embedded (FFOE) bone marrow (BM) aspirates was evaluated as a reliable resource for morphological, immunohistochemical and molecular examinations. One hundred BM aspirates were collected in polypropylene tubes and immediately frozen for 2 h in a deep freezer. Frozen BM was transferred to a cryomold filled with OCT compound and the prepared samples were stored in a deep freezer. Histological examination and immunohistochemical staining, polymerase chain reaction (PCR), sequencing and reverse transcription (RT)-PCR were performed to evaluate the quality of the FFOE BM sections in 10% of randomly selected samples. FFOE BM sections revealed better morphologies than paraffin-embedded clot sections in haematoxylin and eosin staining because mature erythrocytes were removed during the staining process in frozen BM sections. Immunohistochemical staining for CD34 revealed excellent staining quality and oil red O staining showed that fat vacuoles in cells were well preserved. The quality of genomic DNA in FFOE BM sections was suitable for obtaining about 2000 bp PCR product for the human leucocyte antigen-A locus followed by direct sequencing of the sample, and the quality of total RNA was suitable for detection of BCR-ABL fusion transcript. FFOE BM aspirates are a reliable resource for various laboratory tests of diagnostic and research arenas. PMID- 19298223 TI - The oral iron chelator deferasirox represses signaling through the mTOR in myeloid leukemia cells by enhancing expression of REDD1. AB - To evaluate the effect of deferasirox in human myeloid leukemia cells, and to identify the molecular pathways responsible for antiproliferative effects on leukemia cells during chelation therapy, we performed gene expression profiling to focus on the pathway involved in the anticancer effect of deferasirox. The inhibitory concentration (IC50) of deferasirox was 17-50 microM in three human myeloid cell lines (K562, U937, and HL60), while those in fresh leukemia cells obtained from four patients it varied from 88 to 172 microM. Gene expression profiling using Affymerix GeneChips (U133 Plus 2.0) revealed up-regulation of cyclin-dependent kinase inhibitor 1A (CDKN1A) encoding p21CIP, genes regulating interferon (i.e. IFIT1). Pathways related to iron metabolism and hypoxia such as growth differentiation factor 15 (GDF-15) and Regulated in development and DNA damage response (REDD1) were also prominent. Based on the results obtained from gene expression profiling, we particularly focused on the REDD1/mTOR (mammalian target of rapamycin) pathway in deferasirox-treated K562 cells, and found an enhanced expression of REDD1 and its down-stream protein, tuberin (TSC2). Notably, S6 ribosomal protein as well as phosphorylated S6, which is known to be a target of mTOR, was significantly repressed in deferasirox-treated K562 cells, and REDD1 small interfering RNA restored phosphorylation of S6. Although iron chelation may affect multiple signaling pathways related to cell survival, our data support the conclusion that REDD1 functions up-stream of tuberin to down regulate the mTOR pathway in response to deferasirox. Deferasirox might not only have benefit for iron chelation but also may be an antiproliferative agent in some myeloid leukemias, especially patients who need both iron chelation and reduction of leukemia cells. PMID- 19298224 TI - Therapeutic potential of proapoptotic molecule Noxa in the selective elimination of tumor cells. AB - The selective elimination of tumor cells by inducing apoptosis is one of the most important issues in cancer therapy. In this context, artificial expression of the p53 tumor-suppressor gene has been an attractive approach and numerous studies have shown its efficacy in combination with other therapies such as radiation or chemotherapy. One of the critical issues for current cancer gene therapy is how to induce apoptosis in cancer cells without affecting normal cells. In the present study, we examined the potential of Noxa, a BH3-only protein with proapoptotic activity that functions downstream of the p53-mediated apoptotic pathway, to selectively induce apoptosis in tumor cells. We found that upon infection of a recombinant adenovirus contrived to express the Noxa gene, apoptosis was induced in vitro in several human breast cancer cell lines, but not in normal mammary epithelial cell lines. Furthermore, intratumoral injection of the Noxa-expressing adenovirus resulted in marked shrinkage of the transplanted tumor derived from breast cancer cells without any notable adverse effect on the surrounding normal tissue. In contrast, the expression of Puma, another BH3-only protein that also functions downstream of the p53 pathway, induced apoptosis in both cancer and normal cells. Thus, our results suggest a mechanism wherein Noxa, but not Puma, selectively induces apoptosis in human tumor cells. These data provide a new prospect for cancer therapy by the Noxa-mediated selective elimination of malignant cells. PMID- 19298225 TI - The significance of organizational factors for the incidence of bullying. AB - It is frequently assumed that a poor psychosocial working environment will create conditions that encourage bullying. However, few studies have examined this assumption while comparing work environment ratings of bullied and non-bullied employees who work in the same organization and/or department. The objective of this study was to examine the relationship between organizational factors and the incidence of acts of bullying based on two different approaches: the first by comparing bullied with no-bullied, the second by comparing departments with widespread bullying with departments with little bullying. The study was a part of a general survey study of the work environment and employee well-being in 12 different local social security offices. A total of 898 persons participated in the study (a response rate of 88%). First, the results showed a clear relationship between bullying and fear of organizational change; secondly, weak, but significant, correlations between bullying and other organizational factors; thirdly, the subsequent analyses compared departments in which bullying were most widespread with the rest of the departments. The results supported the hypothesis that departments that suffer from much bullying also have a poorer psychosocial work environment, results that support the assumption that organizational factors such as changes in one's position, pressure of work, performance demands, autocratic management and role conflict and lack of role clarity, as well as a poor social climate can contribute to the emergence of higher incidences of bullying. PMID- 19298226 TI - A first French assessment of population exposure to tetrachloroethylene from small dry-cleaning facilities. AB - Used as a solvent in the dry-cleaning industry, tetrachloroethylene (C(2)Cl(4)) can be a pollutant of residential indoor air, which can cause long-term harmful exposures because of its neurotoxicity and probable carcinogenicity. In France, dry-cleaning facilities are integrated in urban environments (shopping malls, residential buildings) and can contribute to C(2)Cl(4) exposure for customers and residents. This exploratory work presents the results from five studies carried out in one shopping mall and four residential buildings housing a dry-cleaning facility. These studies involved dry-cleaning machines fitted with a Carbon Adsorber and unfitted, with or without Air Exhaust System. Samples were collected in the cleaning facilities and in the apartments located above with passive samplers allowing measurement of time-integrated concentrations on a 7 days sampling period. It has obviously shown the degradation of indoor air quality in these environments and underlined the contributing role of the machine technology and ventilation system on the amount of released C(2)Cl(4) in the indoor air. To temper these results, it must be pointed out that some parameters (building insulation, amount of solvent used...) which would influence C(2)Cl(4) fugitive release have not been quantified and should be looked at in further studies. PRACTICAL IMPLICATIONS: In France, dry-cleaning facilities are frequently integrated in urban environments (large shopping malls or residential buildings) and can significantly contribute to tetrachloroethylene (C(2)Cl(4)) population exposure. The amount of fugitive releases in these environments depends on several parameters such as the dry-cleaning machine technology (fitted or unfitted with a carbon adsorber) and the ventilation (air exhaust system). To reduce C(2)Cl(4) exposure in residential buildings and other indoor environments with on-site dry cleaners, carbon adsorber unequipped machine should be replaced by newer technology and dry cleaners should be equipped with mechanical air exhaust systems. PMID- 19298227 TI - Nasal hyperresponders and atopic subjects report different symptom intensity to air quality: a climate chamber study. AB - Short-term exposure to dust and dust added with beta-(1,3)-d-glucan or aldehydes may cause sensory reactions. In random order, we exposed 36 volunteers in a climate chamber to clean air, office dust, dust with glucan, and dust with aldehydes. Three groups of subjects were exposed, eleven were non-atopic with nasal histamine hyperreactivity, 13 were non-atopic, and 12 were atopic. Subjective ratings of symptoms and general health were registered four times during four 6-h exposure sessions. Six symptom intensity indices were constructed. The nasal hyperreactive group had a high and time-dependent increase of mucous membrane irritations, whereas the atopic group had a low and stable rate of irritations with exposure time, close to the reference group (P = 0.02 for differences between the groups with respect to time under exposure for Weak Inflammatory Responses and P = 0.05 for Irritative Body Perception, significance mainly because of the nasal hyperreactive group). Exposure to dust, with or without glucan or aldehydes, showed increased discomfort measured by the index for Constant Indoor Climate, and dust with glucan had a similar effect for the index for Lower Respiratory Effects. For Psychological and Neurological Effects these were dependent on group affiliation, thus preventing a uniform statement of exposure effects for all three investigated groups. PRACTICAL IMPLICATIONS: Opportunities for identifying persons with high or low sensitivity to low-level exposures are important in preventive medicine and will reduce intra-group variability and thus increase the power of experimental and epidemiological studies searching for correlations between exposures and health effects. The contrast between nasal hyperreactive on one side and atopic and reference subjects on the other side is particularly important. The atopic group indicated a non-homogenous reaction depending on their hyperreactive status, a finding that could be important but needs further confirmation. PMID- 19298228 TI - Low home ventilation rate in combination with moldy odor from the building structure increase the risk for allergic symptoms in children. AB - There are consistent findings on associations between asthma and allergy symptoms and residential mold and moisture. However, definitions of 'dampness' in studies are diverse because of differences in climate and building construction. Few studies have estimated mold problems inside the building structure by odor assessments. In a nested case-control study of 400 Swedish children, observations and measurements were performed in their homes by inspectors, and the children were examined by physicians for diagnoses of asthma, eczema, and rhinitis. In conclusion, we found an association between moldy odor along the skirting board and allergic symptoms among children, mainly rhinitis. No associations with any of the allergic symptoms were found for discoloured stains, 'floor dampness' or a general mold odor in the room. A moldy odor along the skirting board can be a proxy for hidden moisture problem inside the outer wall construction or in the foundation construction. There are indications that such dampness problems increase the risk for sensitization but the interpretation of data in respect of sensitization is difficult as about 80% of the children with rhinitis were sensitized. Furthermore, low ventilation rate in combination with moldy odor along the skirting board further increased the risk for three out of four studied outcomes, indicating that the ventilation rate is an effect modifier for indoor pollutants. PRACTICAL IMPLICATIONS: This study showed that mold odor at the skirting board level is strongly associated with allergic symptoms among children. Such odor at that specific place can be seen as a proxy for some kind of hidden moisture or mold problem in the building structure, such as the foundation or wooden ground beam. In houses with odor along the skirting board, dismantling of the structure is required for an investigation of possible moisture damage, measurements, and choice of actions. In homes with low ventilation in combination with mold odor along the skirting board, there was even a higher risk of health effects. This emphasizes the need for the appropriate remediation as this is an ever increasing problem in poorly ventilated houses that are damp. PMID- 19298229 TI - Possible role for TRPV1 in neomycin-induced inhibition of visceral hypersensitivity in rat. AB - Transient receptor ion channel 1 (TRPV1) is a nociceptor involved in visceral hypersensitivity. Aminoglycosides like neomycin are not only potent antibiotics but in vitro data suggest that neomycin also acts as a TRPV1-antagonist and alleviates somatic pain responses. To what extent neomycin reduces visceral hypersensitivity remains unknown. Therefore, we aimed to investigate whether neomycin can inhibit in vivo TRPV1-dependent hypersensitivity responses in two rat models of visceral pain. In the first model rats were pretreated with intraperitoneal (i.p.) capsazepine, the selective TRPV1 antagonist SB-705498, neomycin or vehicle alone and 30 min later instilled with intracolonic TRPV1 activating capsaicin. Likewise, rats were pretreated with 10 days oral neomycin and then subjected to intracolonic capsaicin. The visceromotor response (VMR) to distension was measured before and after capsaicin application. In addition, the VMR to distension was measured in adult maternal separated rats before and after acute stress. Before the 2nd distension protocol these rats were treated with i.p. neomycin, amoxycillin or vehicle alone. Our results showed that capsaicin administration induced an enhanced VMR to distension that was prevented by i.p. capsazepine, SB-705498 and neomycin. Oral neomycin treatment changed bacterial faecal content but could not inhibit capsaicin induced visceral hypersensitivity. In maternal separated rats acute stress induced an enhanced response to distension that was reversed by i.p. neomycin, but not amoxycillin. These data indicate that (i.p.) neomycin can inhibit visceral hypersensitivity to distension in a nonbactericidal manner and suggest that TRPV1-modulation may be involved. PMID- 19298230 TI - Inhibitory effect of oxytocin on accelerated colonic motility induced by water avoidance stress in rats. AB - Recent studies have indicated that brain and gut activities are interrelated and exposure to several stressors, such as water-avoidance stress, stimulates the motor function of the gut through corticotropin-releasing factor (CRF)-signalling pathways in the brain. Central oxytocin is known to attenuate stress responses, including CRF expression in the brain. Here, we examined whether central oxytocin attenuated the acceleration of colonic motility induced by water-avoidance stress. A force transducer was attached to the distal colon of male rat, and the colonic motility and faecal pellet output were recorded while the rats were exposed to water-avoidance stress. Intracerebroventricular (i.c.v.) injections of oxytocin (5, 50 and 500 pmol) and the oxytocin receptor antagonist tocinoic acid (25 microg) were administered before exposure to water-avoidance stress, and the effect of oxytocin on colonic motor function was determined. Centrally administered oxytocin inhibited the accelerated colonic motility induced by water avoidance stress. The effective dose ranged between 5 and 50 pmol on i.c.v. injection. Oxytocin also decreased the number of CRF-positive cells in the paraventricular nucleus and corticosterone release. The inhibitory effect of oxytocin on accelerated colonic motility was blocked by pretreatment with oxytocin receptor antagonist. Furthermore, centrally administered tocinoic acid enhanced the acceleration of colonic motility. These results suggested that endogenous central oxytocin may contribute to the regulation of colonic function and inhibit the brain CRF-signalling pathways targeting the gut, resulting in the inhibition of stress-induced colonic contractions. PMID- 19298231 TI - Probiotics during weaning reduce the incidence of eczema. AB - A reduced microbial load early in life has been suggested to be linked to the increasing prevalence of allergic diseases in the industrialized world. Some studies have indicated that probiotics may be effective in the prevention of eczema. In vitro studies indicate that probiotics have immunomodulatory effects. In the present study, we evaluated the effects of feeding Lactobacillus F19 during weaning on the incidence of eczema and Th1/Th2 balance. In a double-blind, placebo-controlled randomized intervention trial, infants were fed cereals with (n = 89) or without Lactobacillus F19 (n = 90) from 4 to 13 months of age. We assessed the cumulative incidence of eczema at 13 months of age. The ratio of interferon-gamma (IFN-gamma) to interleukin 4 (IL4) mRNA expression levels in polyclonally stimulated peripheral blood T cells was used as a proxy for immune balance. Total and specific IgE serum levels were also assessed. The cumulative incidence of eczema at 13 months was 11% (4-17%, 95% CI) and 22% (13-31%, 95% CI) in the probiotic and placebo groups, respectively (p < 0.05). The number needed to treat was 9 (6.5-11.5, 95% CI). At 13 months of age, the IFN-gamma/IL4 mRNA ratio was higher in the probiotic compared with the placebo group (p < 0.05). In contrast, there were no differences between groups in serum concentrations of IgE. In summary, feeding Lactobacillus F19 during weaning could be an effective tool in the prevention of early manifestation of allergy, e.g., eczema. The higher Th1/Th2 ratio in the probiotic compared with the placebo group suggests enhancing effects of Lactobacillus F19 on the T cell-mediated immune response. PMID- 19298232 TI - Importance of medication adherence from the ONTARGET and TRANSCEND points of view. AB - Medication adherence is crucial to improve the prognosis of patients with hypertension. Importantly, the variables associated with antihypertensive medication withdrawal do not occur immediately after the cessation, but likely in the following years. Nevertheless, since the current randomized clinical trials are limited to a short or mid follow-up, the results provided from these studies cannot exactly reflect the importance of the therapy discontinuation. We discuss about two recent examples, the ONTARGET and the TRANSCEND trials, in which the inclusion of the discontinuation rates in the primary analysis change the results of these studies and subsequently, the interpretation of the data. PMID- 19298233 TI - Dextropropoxyphene withdrawal from a French university hospital: impact on analgesic drug consumption. AB - Dextropropoxyphene is a weak opioid analgesic, widely used as a step 2 analgesic (according to WHO classification) in combination with peripheral analgesics, mainly paracetamol. Recent data have underlined its poor analgesic efficacy (in comparison with paracetamol), risks of serious adverse drug reactions (i.e. hepatic reactions, hallucinations, abuse, withdrawal symptoms, hypoglycaemia), possible lethality after overdose, its risk of accumulation in patients with renal failure or in elderly people and some pharmacokinetic insufficiencies (i.e. different half-lives for dextropropoxyphene and paracetamol). Taking into account these data, the drug committee of the Toulouse University Hospital (France) decided to withdraw dextropropoxyphene from the hospital formulary since 1 June 2005. The aim of our study was to investigate the consequences of this withdrawal by comparing use of analgesic drugs in Toulouse University Hospital before (2004) and after (2006) dextropropoxyphene withdrawal (using defined daily dose for 1000 hospitalization-days as the unit measure). Before withdrawal, dextropropoxyphene (in combination with paracetamol) was the second most used analgesic drug after paracetamol alone. After dextropropoxyphene withdrawal, total consumption of analgesic drugs decreased by 4.6% (2006 vs. 2004). There was a 28% decrease in consumption of step 2 analgesics [with an increase in oral tramadol and a slight decrease in codeine (in combination with paracetamol)]. During the same period, step 1 analgesic consumption increased by 11% (mainly paracetamol) and that of step 3 analgesics slightly decreased (-8%). These results show that dextropropoxyphene withdrawal was not associated with a marked switch in prescriptions towards other analgesic drugs. This paper underlines the interest of a hospital-based drug committee to promote rational drug use. Finally, the present data allow us to discuss putative misuse of dextropropoxyphene. PMID- 19298234 TI - Y27632, a Rho-activated kinase inhibitor, normalizes dysregulation in alpha1 adrenergic receptor-induced contraction of Lyon hypertensive rat artery smooth muscle. AB - RhoA-activated kinase (ROK) is involved in the disorders of smooth muscle contraction found in hypertension model animals and patients. We examined whether the alpha1-adrenergic receptor agonist-induced ROK signal is perturbed in resistance small mesentery artery (SMA) of Lyon genetically hypertensive (LH) rats, using a ROK antagonist, Y27632. Smooth muscle strips of SMA and aorta were isolated from LH and Lyon normotensive (LN) rats. After Ca(2+)-depletion and pre treatment with phenylephrine (PE), smooth muscle contraction was induced by serial additions of CaCl(2). In LH SMA Ca(2+) permeated cells to a lesser extent as compared with LN SMA, while CaCl(2)-induced contraction of LH SMA was greater than that of LN SMA, indicating a higher ratio of force to Ca(2+) in LH SMA contraction (Ca(2+) sensitization). No hyper-contraction was observed in LH aorta tissues. Treatment of LH SMA with Y27632 restored both Ca(2+) permeability and Ca(2+)-force relationship to levels seen for LN SMA. In response to PE stimulation, phosphorylation of CPI-17, a phosphorylation-dependent myosin phosphatase inhibitor protein, and MYPT1 at Thr853, the inhibitory phosphorylation site of the myosin phosphatase regulatory subunit, was increased in LN SMA, but remained unchanged in LH SMA. These results suggest that the disorder in ROK-dependent Ca(2+) permeability and Ca(2+)-force relationship is responsible for LH SMA hyper-contraction. Unlike other hypertensive models, the ROK-induced hyper-contractility of LH SMA is independent of MYPT1 and CPI-17 phosphorylation, which suggests that ROK-mediated inhibition of myosin phosphatase does not affect SMA hyper-contractility in LH SMA cells. PMID- 19298235 TI - Nicotinic acetylcholine receptors mediate the hypnotic and analgesic effects of emulsified inhalation anesthetics. AB - This study was designed to investigate the role of nicotinic acetylcholine receptors (nAChRs) in hypnosis and analgesia induced by emulsified inhalation anesthetics. After having established the mice model of hypnosis and analgesia by intraperitoneal injections of appropriate doses of enflurane, isoflurane or sevoflurane, we intracerebroventricularly or intrathecally injected different doses of nicotine and then observed the effects on the sleeping time using awaken test and the pain threshold in hot-plate test (HPPT) using hot-plate test. In the awaken test, 10, 20 and 40 microg of nicotine (intracerebroventricularly) significantly decreased the sleeping time of the mice treated with the three emulsified inhalation anesthetics mentioned above (P < 0.05 or 0.01). In the HPPT, 5, 10 and 15 microg of nicotine (intrathecally) did not affect the HPPT in conscious mice (P > 0.05); in contrast, 5, 10 and 15 microg of nicotine (intrathecally) significantly decreased the HPPT of the mice treated with emulsified inhalation anesthetics (P < 0.05 or 0.01). The data presented in this study suggest that nAChRs may be important targets for the hypnotic and analgesic effects induced by emulsified enflurane, isoflurane and sevoflurane. PMID- 19298236 TI - Slow release properties and liver-targeting characteristics of methotrexate erythrocyte carriers. AB - To study the releasing properties and tissue-targeting characteristics of methotrexate-loaded red blood cells (MTX-RBCs), pharmacokinetics and tissue distributions of intravenous injected MTX-RBCs and free MTX were compared. MTX RBCs were made from rat erythrocytes using a hypertonic method. After i.v. injection of MTX-RBCs or free MTX to rats, both plasma and tissue homogenate samples at each time-point were collected and analyzed by RP-HPLC. From this data, pharmacokinetic parameters and tissue distributions of MTX were obtained. MTX-RBCs were successfully produced by the hypertonic method. After i.v. injection, MTX-RBCs displayed more than three times longer half-life and MRT than free MTX, and the velocity of MTX clearance from plasma was much slower. The ratio of area under the concentration time curve (AUC)(tissue) to AUC(plasma) in the liver was clearly higher than that for other organs after MTX-RBCs administration; MRT in the liver was also longer. This study has demonstrated that the hypertonic method for making MTX-RBCs has led to a preparation with slow release properties as well as liver-targeting characteristics in rats. This approach offers considerable potential for the treatment of tumors in liver, which would merit further investigation. PMID- 19298237 TI - Quantitative analysis of the scientific literature on acetaminophen in medicine and biology: a 2003-2005 study. AB - This study quantifies the utilization of acetaminophen in life sciences and clinical medicine using bibliometric indicators. A total of 1626 documents involving acetaminophen published by 74 countries during 2003-2005 in the Thompson-Scientific Life sciences and Clinical Medicine collections were identified and analyzed. The USA leads in the number of publications followed by the UK, and industrialized countries, including France, Japan and Germany; the presence of countries such as China, India and Turkey among the top 15 countries deserves to be noticed. The European Union stands as a comparable contributor to the USA, both in terms of number of publications and in terms of profile of papers distributed among subcategories of Life Sciences and Clinical Medicine disciplines. All documents were published in 539 different journals. The most prolific journals were related to pharmacology and/or pharmaceutics. All aspects of acetaminophen (chemistry, pharmacokinetics, metabolism, etc.) were studied with primary interest for therapeutic use (42%) and adverse effects (28%) comprising a large part of publications focusing on acetaminophen hepatotoxicity. This quantitative overview provides as to the interest of the scientific community in this analgesic and completes the various review documents that regularly appear in the scientific literature. PMID- 19298238 TI - Hepatoprotective effects of misoprostol and silymarin on carbon tetrachloride induced hepatic damage in rats. AB - The aim of this study was to investigate the effect of misoprostol, silymarin or the co-administration of misoprostol + silymarin on the carbon tetrachloride (CCl(4))-induced hepatic injury in rats. Misoprostol (10, 100, 1000 microg/kg), silymarin (25 mg/kg) or misoprostol (100 microg/kg) + silymarin (25 mg/kg) was given once daily orally simultaneously with CCl(4) and for 15 days thereafter. The results showed that misoprostol (10, 100 or 1000 microg/kg) conferred significant protection against the hepatotoxic actions of CCl(4) in rats, reducing serum alanine aminotransferase (ALT) levels by 24.7%, 42.6% and 49.4%, respectively compared with controls. Misoprostol, given at 100 or 1000 microg/kg, decreased aspartate aminotransferase (AST) by 28 and 43.6% and alkaline phosphatase (ALP) by 19.3% and 53.4% respectively. Meanwhile, silymarin reduced ALT, AST and ALP levels by 62.7%, 66.1% and 65.1% respectively. The co administration of misoprostol (100 microg/kg) and silymarin (25 mg/kg) resulted in 61.4%, 66.1% and 57.5% reduction in ALT, AST and ALP levels respectively. Histopathological alterations and depletion of hepatocyte glycogen and DNA content by CCl(4) were markedly reduced after treatment with misoprostol, silymarin or misoprostol + silymarin. Image analysis of liver specimens revealed a marked reduction in liver necrosis; area of damage: 32.4%, 24% and 10.2% after misoprostol (10, 100 or 1000 microg/kg), 7.2% after silymarin and 10.9% after treatment with misoprostol 100 microg/kg + silymarin, compared with CCl(4) control group (46.7%). These results indicate that treatment with misoprostol protects against hepatocellular necrosis induced by CCl(4). This study suggests a potential therapeutic use for misoprostol in liver injury. PMID- 19298239 TI - Infectious complications after allogeneic stem cell transplantation: incidence in matched-related and matched-unrelated transplant settings. AB - BACKGROUND: Bacterial, viral, and fungal pathogens frequently cause severe, life threatening infections in immunocompromised patients after allogeneic hematopoietic stem cell transplantation (SCT). OBJECTIVE: To compare the frequency of infections in patients with matched-related (Group A) or with human leukocyte antigen (HLA)-matched-unrelated donors (Group B). PATIENTS AND METHODS: Patients treated at our transplantation unit between April 2004 and April 2005 were enrolled into this analysis. Documentation comprised demographic data, conditioning treatment, stem cell source, clinical course, as well as microbiological and clinical data and mortality. RESULTS: We analyzed 59 patients, 22 in Group A and 37 in Group B. Both groups were well balanced regarding demographic data. Diagnoses were acute myeloid leukemia (30 of 59 patients, 50.8%), multiple myeloma (15.2%), acute lymphoblastic leukemia (11.9%), and chronic myeloid leukemia (10.2%). Patients in Group A developed infections in 95.5% of the cases compared with 97.3% in patients in Group B. Most frequently detected pathogens were Staphylococcus species, human herpesvirus-6, and Epstein Barr virus. Three proven fungal infections were detected in Group A compared with 9 proven fungal infections in Group B. Lung infiltrations were observed in equivalent incidence in both groups. Two years after transplantation, 55.9% of patients were alive (Group A: 68.2%; Group B: 48.6%, not significant). CONCLUSION: Allogeneic SCT from HLA-matched-unrelated donors does not have a higher infection risk than patients transplanted from matched-related donors. PMID- 19298240 TI - Mycobacterium abscessus in cystic fibrosis lung transplant recipients: report of 2 cases and risk for recurrence. AB - Mycobacterium abscessus is increasingly recognized as an important pathogen in some individuals with advancing lung disease related to cystic fibrosis (CF). Because of its resistance to antimicrobial agents and virulence, its presence in the lungs of potential lung transplant recipients can be problematic. We present 2 cases of individuals with CF in whom M. abscessus was present in the preoperative sputum cultures. The organism manifested different degrees of invasiveness in the 2 cases after transplantation with different outcomes, suggesting an approach to future candidates for lung transplantation that may be of clinical significance to their physicians and surgeons. PMID- 19298241 TI - Lung cavitation due to Mycobacterium xenopi in a renal transplant recipient. AB - Mycobacterium xenopi is an unusual pathogen and few such cases have been reported in the literature. We report the case of a patient with a sirolimus-based immunosuppressive regimen, who developed lung cavitation. M. xenopi was isolated from the sputum. The patient was treated initially with rifampicin, isoniazid, and pyrazinamide; levofloxacin was added to the treatment regimen after M. xenopi was demonstrated. A possible relationship between sirolimus and M. xenopi infection has been postulated, probably due to the combination of pulmonary toxicity and cellular immunosuppression of rapamycin. PMID- 19298242 TI - Differential gene expression profiles of whole lesions from patients with oral lichen planus. AB - BACKGROUND: The molecular biological properties of oral lichen planus (OLP) are largely unknown. The aim of this study was to identify the genes responsible for its pathogenesis at the genome scale using DNA microarray technology. METHODS: The RNA samples extracted from the specimens of nine OLP patients and nine controls were analyzed with Affymetrix GeneChip. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis was applied to confirm GeneChip results. RESULTS: A total of 985 differentially expressed genes (629 up-regulated and 356 down-regulated) were identified. These genes were involved in many function classifications and biochemical pathways. The results of quantitative RT PCR analysis of FOXP3, VEGFA, ANGPT1, MMP1, and SCGB2A2 were consistent with their changes demonstrated by GeneChip. CONCLUSIONS: This study showed the gene expression profiles of OLP, which were quite distinct from that of healthy controls. These results presented a global view of physiopathologic processes in lesions, which will give important clues to understand pathogenesis and identify new therapeutic targets of OLP. PMID- 19298243 TI - Nuclear BCL10 in primary Sjogren's syndrome. AB - BACKGROUND: The events following triggering of antigen receptors and subsequent activation of the transcription factor nuclear factor kappa B (NFkappaB) need to be carefully controlled to prevent abnormal immune responses. BCL10 links the antigen receptor to NFkappaB. The aim of this study was to determine the expression pattern of BCL10 and NFkappaB in minor salivary gland infiltrates of patients with primary Sjogren's syndrome (pSS). METHODS: Minor salivary glands from patients with primary SS (n = 17) and sicca controls (n = 4) were evaluated by single and double immunohistochemistry and immunofluorescence for confocal microscopy. BCL10 and NFkappaB-p65 expression were evaluated in the infiltrating lymphocytes. Ectopic germinal centers (GCs) were investigated by CD21. Tonsil, lymph node and lymphoma tissue were used as positive controls. RESULTS: BCL10 nuclear positive cells were observed in focal lymphocytic infiltrates in the investigated minor salivary glands and were not restricted to patients with ectopic GCs. By double-staining, some of the BCL10 nuclear positive cells were identified as B cells. There was, however, no constitutive activation of NFkappaB as depicted by the exclusive cytoplasmic expression of p65 in the infiltrating lymphocytes in the pSS. CONCLUSION: Nuclear expression of BCL10 in infiltrating lymphocytes was a common occurrence in pSS minor salivary glands indicating it as a possible marker of autoimmune induced chronic inflammation. There was, however, no constitutive activation of NFkappaB. PMID- 19298244 TI - Peroxiredoxin I is overexpressed in oncocytic lesions of salivary glands. AB - BACKGROUND: Oncocytic lesions, particularly frequent in the salivary glands, are characterized by cells with an atypical accumulation of mitochondria. This accumulation has been recognized as a compensatory mechanism to intrinsic functional defects of these organelles, resulting in energy production impairment and increased generation of reactive oxygen species (ROS), including hydrogen peroxide (H(2)O(2)). Peroxiredoxin I (Prx I) is a H(2)O(2) scavenging protein and the expression of its yeast homolog was reported to be influenced by mitochondrial function. METHODS: In this study, we evaluated Prx I expression in oncocytic lesions of salivary glands by immunohistochemistry. RESULTS: Our results showed that Prx I is overexpressed in oncocytes regardless of the salivary gland lesion where they appear. CONCLUSIONS: These results suggest that Prx I expression in oncocytes is related to its ability to decompose mitochondrial-derived H(2)O(2) and that it could provide to the cells a protective role in an environment that, by continuously producing potential DNA damaging ROS, predisposes to genome instability and cellular transformation. PMID- 19298245 TI - A mechanism for thrombin-dependent lung metastasis in patients with osteosarcoma. PMID- 19298246 TI - Oligoclonal TRBV gene usage among CD8(+) T cells in monoclonal B lymphocytosis and CLL. PMID- 19298247 TI - Metastatic melanoma in the marrow: a black and white diagnosis. PMID- 19298248 TI - Military leaders and followers--do they have different decision styles? AB - There are five different, although not mutually exclusive, styles exhibited by military officers when making decisions: rational, intuitive, dependent, avoidant, and spontaneous (Scott & Bruce, 1995). The purpose was to investigate if elected leaders of military planning teams had a different configuration of decision-making styles than their team members. Participants were 98 army captains organized in 16 brigade-level planning teams. The results indicate that team leaders tended to be more spontaneous and less rational, dependent and avoidant in their style configuration than their team members. One possible explanation is that the style configuration exhibited by many of the elected leaders comes through to others as forcefulness and decisiveness and that such a profile is in line with a general leadership culture. The results also provide support for the General Decision-Making Style inventory as a measurement of decision styles, because they suggest that the self-reports coincide with displayed, observable behavior. PMID- 19298249 TI - Gambling behavior and gambling problems in Norway 2007. AB - In Norway, legalized gambling is pervasive, easily accessible and socially accepted, particularly the participation in national lotteries. We conducted a stratified probability sample study during January-March 2007 (age group 16-74 years, N = 3,482, response rate 36.1%) to assess gambling behavior and prevalence of problematic gambling by the NORC Diagnostic Screen (NODS). Overall, 67.9% of the study population had been engaged in past-year gambling and 0.7% were past year problematic gamblers (NODS score 3+). Male gender, low educational level, single marital status, and being born in a non-Western country were variables positively associated with problematic gambling. Past-year problematic gamblers indicated the slot machine (34.6%) and Internet gambling (26.9%) as the most important games, while most non-problematic gamblers pointed out the lotteries (62.3%) as the most important game. The prevalence of self-reported sleeping disorders, depression and other psychological impairments was significantly higher among problematic gamblers. Gambling problems affect people's lives in multiple ways. PMID- 19298250 TI - Self-esteem and interpersonal functioning in psychiatric outpatients. AB - This study explored associations between self-esteem and interpersonal functioning in a one-year clinic cohort of psychiatric outpatients (n= 338). At intake, patients completed questionnaires measuring self-esteem, interpersonal problems, interpersonal style, and general symptomatic distress. They were also diagnosed according to the ICD-10. Interpersonal behaviour was measured along the agency and communion dimensions of the Inventory of Interpersonal Problems Circumplex [IIP-C]. The results show that lower self-esteem was associated with higher levels of interpersonal problems in general. Further, lower self-esteem was first and foremost linked to frustrated agentic motives, as measured by the IIP-C. Hence, the study concludes that fostering patient agency should be considered as an important goal in psychotherapy. Furthermore, the analyses revealed an interaction effect of agency and communion on self-esteem, indicating a need for balancing the two motive dimensions. Finally, some questions are raised concerning the interpretation of the IIP-C subscales in general. PMID- 19298251 TI - Successful kidney transplantation from a donation after cardiac death donor with acute renal failure and bowel infarction using extracorporeal support. AB - As a result of the ever widening disparity between organ supply and demand, a resurgence of interest has occurred in kidney recovery from donation after cardiac death (DCD) donors. New techniques of in situ extracorporeal support offer the potential to reduce warm ischemic injury and optimize donor management prior to organ recovery. In addition, preliminary outcomes using kidneys from selected deceased donors with rising serum creatinine levels have been promising. However, contraindications to successful organ donation and transplantation may include the presence of abdominal compartment syndrome, generalized bowel infarction, refractory shock with profound metabolic and lactic acidosis, and acute anuric renal failure, particularly in the setting of DCD. We report herein the successful recovery and transplantation of kidneys from an unstable donor with the above constellation of conditions in the setting of extracorporeal support after declaration of death by asystole. PMID- 19298252 TI - A simple and reliable method to blood type monkeys using serum samples. AB - Monkeys are frequently used in experimental transplantation research because of their physical traits and availability. As ABO incompatibility may result in humoral injury, it is important to identify the ABO blood typing of monkeys before transplantation. However, monkeys lack expression of ABH antigens on red blood cells, which makes accurate determination of the blood type difficult. The gel agglutination assay has been widely used as a routine blood grouping test clinically for more than 10 years. In this study, we evaluated the efficacy and the interference factors of using the gel system (including the direct gel system and the reverse gel system) for ABO typing in rhesus monkeys (n = 38) and cynomolgus monkeys (n = 26). Immunohistochemistry assay was used to obtain the accurate blood type data of monkeys. The results revealed that the direct gel system was ineffective in blood typing of monkeys, whereas the reverse gel system assay, which is based on preabsorbed serum, provided reproducible results that were confirmed by histologic analysis. We conclude that the reverse gel system assay with use of preabsorbed serum is a simple and reliable method for ABO typing of monkeys. PMID- 19298253 TI - Mitogen-activated protein kinase (MAPK) pathway mediates the oestrogen-like activities of ginsenoside Rg1 in human breast cancer (MCF-7) cells. AB - BACKGROUND AND PURPOSE: The present study was designed to determine how ginsenoside Rg1, an active ingredient in ginseng root, exerts its oestrogenic effects. We hypothesize that Rg1 may exert oestrogen-like actions in MCF-7 cells by activating the mitogen-activated protein kinase (MAPK) pathway in a ligand independent manner. EXPERIMENTAL APPROACH: MCF-7 cells were co-incubated with the MAPK inhibitor PD98059 to determine whether the stimulant effects of Rg1 on cell proliferation, the induction of IGF-IR and pS2, the functional transactivation of oestrogen receptor-alpha (ERalpha), as well as ERalpha phosphorylation are dependent on MAPK. The time-dependent responses of mitogen-activated protein kinase kinase (MEK) and extracellular signal-regulated protein kinase (ERK) to Rg1 in MCF-7 cells were studied. The responses of MEK phosphorylation to Rg1 in oestrogen receptor (ER)-negative HEK293 cells were also determined. The effects of Rg1 on cell proliferation and IGF-IR protein expression were studied in the presence of tyrosine kinase inhibitor genistein to elucidate the involvement of tyrosine kinase in mediating these effects. KEY RESULTS: The oestrogenic effects of Rg1 in MCF-7 cells were abolished in the presence of PD98059. Rg1 could induce MEK protein expression and the phosphorylation level of MEK and ERK significantly in a time- and dose-dependent manner. Rg1 activated MEK phosphorylation in ER negative HEK293 cells in a time- and dose-dependent manner. Rg1 induction of cell proliferation and IGF-IR protein expression was abolished by co-treatment with genistein. CONCLUSIONS AND IMPLICATIONS: Taken together, these results show that the MAPK pathway is involved in mediating the oestrogen-like actions of Rg1 in MCF-7 cells and suggest that Rg1 may activate ERalpha via MEK/ERK in a ligand independent manner. PMID- 19298254 TI - Loss of Ca-mediated ion transport during colitis correlates with reduced ion transport responses to a Ca-activated K channel opener. AB - BACKGROUND AND PURPOSE: Epithelial surface hydration is critical for proper gut function. However, colonic tissues from individuals with inflammatory bowel disease or animals with colitis are hyporesponsive to Cl(-) secretagogues. The Cl(-) secretory responses to the muscarinic receptor agonist bethanechol are virtually absent in colons of mice with dextran sodium sulphate (DSS)-induced colitis. Our aim was to define the mechanism underlying this cholinergic hyporesponsiveness. EXPERIMENTAL APPROACH: Colitis was induced by 4% DSS water, given orally. Epithelial ion transport was measured in Ussing chambers. Colonic crypts were isolated and processed for mRNA expression via RT-PCR and protein expression via immunoblotting and immunolocalization. KEY RESULTS: Expression of muscarinic M(3) receptors in colonic epithelium was not decreased during colitis. Short-circuit current (I(SC)) responses to other Ca(2+)-dependent secretagogues (histamine, thapsigargin, cyclopiazonic acid and calcium ionophore) were either absent or severely attenuated in colonic tissue from DSS-treated mice. mRNA levels of several ion transport molecules (a Ca(2+)-regulated Cl(-) channel, the intermediate-conductance Ca(2+)-activated K(+) channel, the cystic fibrosis transmembrane conductance regulator, the Na(+)/K(+)-ATPase pump or the Na(+)/K(+)/2Cl(-) co-transporter) were not reduced in colonic crypts from DSS treated mice. However, protein expression of Na(+)/K(+)-ATPase alpha1 subunits was decreased twofold during colitis. Activation of Ca(2+)-activated K(+) channels increased I(SC) significantly less in DSS colons compared with control, as did the protein kinase C activator, phorbol 12-myristate 13-acetate. CONCLUSIONS AND IMPLICATIONS: Decreased Na(+)/K(+)-ATPase expression probably contributes to overall epithelial hyporesponsiveness during colitis, while dysfunctional K(+) channels may account, at least partially, for lack of epithelial secretory responses to Ca(2+)-mediated secretagogues. PMID- 19298255 TI - Artemisinin induces doxorubicin resistance in human colon cancer cells via calcium-dependent activation of HIF-1alpha and P-glycoprotein overexpression. AB - BACKGROUND AND PURPOSE: Artemisinin is an antimalarial drug exerting pleiotropic effects, such as the inhibition of the transcription factor nuclear factor-kappa B and of the sarcoplasmic/endoplasmic reticulum Ca(++)-ATPase (SERCA) of P. falciparum. As the sesquiterpene lactone thapsigargin, a known inhibitor of mammalian SERCA, enhances the expression of P-glycoprotein (Pgp) by increasing the intracellular Ca(++) ([Ca(++)](i)) level, we investigated whether artemisinin and its structural homologue parthenolide could inhibit SERCA in human colon carcinoma HT29 cells and induce a resistance to doxorubicin. EXPERIMENTAL APPROACH: HT29 cells were incubated with artemisinin or parthenolide and assessed for SERCA activity, [Ca(++)](i) levels, Pgp expression, doxorubicin accumulation and toxicity, and translocation of the hypoxia-inducible factor, HIF-1alpha. KEY RESULTS: Artemisinin and parthenolide, like the specific SERCA inhibitors thapsigargin and cyclopiazonic acid, reduced the activity of SERCA. They also increased intracellular calcium concentration ([Ca(++)](i)) and Pgp expression and decreased doxorubicin accumulation and cytotoxicity. The intracellular Ca(++) chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, and the inhibitor of calmodulin-dependent kinase II (CaMKII) KN93 prevented these effects. CaMKII is known to promote the phosphorylation and the activation of HIF 1alpha, which may induce Pgp. In HT29 cells, artemisinin and parthenolide induced the phosphorylation of HIF-1alpha, which was inhibited by KN93. CONCLUSIONS AND IMPLICATIONS: Our results suggest that artemisinin and parthenolide may act as SERCA inhibitors and, like other SERCA inhibitors, induce resistance to doxorubicin in human colon cancer cells, via the CaMKII-dependent activation of HIF-1alpha and the induction of Pgp. PMID- 19298256 TI - Neural KCNQ (Kv7) channels. AB - KCNQ genes encode five Kv7 K(+) channel subunits (Kv7.1-Kv7.5). Four of these (Kv7.2-Kv7.5) are expressed in the nervous system. Kv7.2 and Kv7.3 are the principal molecular components of the slow voltage-gated M-channel, which widely regulates neuronal excitability, although other subunits may contribute to M-like currents in some locations. M-channels are closed by receptors coupled to Gq such as M1 and M3 muscarinic receptors; this increases neuronal excitability and underlies some forms of cholinergic excitation. Muscarinic closure results from activation of phospholipase C and consequent hydrolysis and depletion of membrane phosphatidylinositol-4,5-bisphosphate, which is required for channel opening. Some effects of M-channel closure, determined from transmitter action, selective blocking drugs (linopirdine and XE991) and KCNQ2 gene disruption or manipulation, are as follows: (i) in sympathetic neurons: facilitation of repetitive discharges and conversion from phasic to tonic firing; (ii) in sensory nociceptive systems: facilitation of A-delta peripheral sensory fibre responses to noxious heat; and (iii) in hippocampal pyramidal neurons: facilitation of repetitive discharges, enhanced after-depolarization and burst-firing, and induction of spontaneous firing through a reduction of action potential threshold at the axon initial segment. Several drugs including flupirtine and retigabine enhance neural Kv7/M channel activity, principally through a hyperpolarizing shift in their voltage gating. In consequence they reduce neural excitability and can inhibit nociceptive stimulation and transmission. Flupirtine is in use as a central analgesic; retigabine is under clinical trial as a broad-spectrum anticonvulsant and is an effective analgesic in animal models of chronic inflammatory and neuropathic pain. PMID- 19298257 TI - The anorexic agents, sibutramine and fenfluramine, depress GABA(B)-induced inhibitory postsynaptic potentials in rat mesencephalic dopaminergic cells. AB - BACKGROUND AND PURPOSE: Nutrition is the result of a complex interaction among environmental, homeostatic and reward-related processes. Accumulating evidence supports key roles for the dopaminergic neurons of the ventral midbrain in regulating feeding behaviour. For this reason, in the present study, we have investigated the electrophysiological effects of two centrally acting anorexic agents, fenfluramine and sibutramine, on these cells. EXPERIMENTAL APPROACH: Rat midbrain slices were used to make intracellular recordings from dopaminergic neurons of the substantia nigra and the ventral tegmental area. Gamma aminobutyric acid (GABA)-mediated synaptic transmission was assessed from the inhibitory postsynaptic potentials (IPSPs) mediated by GABA(A) and GABA(B) receptors. KEY RESULTS: Fenfluramine and sibutramine reduced, concentration dependently, the GABA(B) IPSPs, without affecting the GABA(A)-mediated potentials. This effect is presynaptic, as postsynaptic membrane responses induced by application of a GABA(B) receptor agonist, baclofen, were not affected by the two drugs. Furthermore, the selective 5-hydroxytriptamine 1B (5-HT(1B)) receptor antagonist, SB216641, blocked the reduction of GABA(B) IPSPs caused by fenfluramine and sibutramine, indicating that the receptor mediating this effect is 5-HT(1B). CONCLUSIONS AND IMPLICATIONS: Two anorexic agents, fenfluramine and sibutramine, induced the activation of 5-HT(1B) receptors located on presynaptic GABAergic terminals, thus reducing the release of GABA. This action can alter the strength of synaptic afferents that modify the activity of dopaminergic neurons, inducing neuronal excitation. Our results reveal an additional mechanism of action for fenfluramine and sibutramine that might contribute to reducing food intake, by influencing the pleasurable and motor aspects of feeding behaviour. PMID- 19298258 TI - Inhibition of inducible NO synthase, cyclooxygenase-2 and interleukin-1beta by torilin is mediated by mitogen-activated protein kinases in microglial BV2 cells. AB - BACKGROUND AND PURPOSE: Traditionally, the stem and root bark of Ulmus davidiana var. japonica (Ulmaceae) have been known to be anti-inflammatory in Korea. Anti inflammatory effects of torilin, isolated from this plant and the underlying mechanisms were examined by using lipopolysaccharide (LPS)-stimulated microglial BV2 cells. EXPERIMENTAL APPROACH: The cells were treated with torilin prior to LPS exposure and the effects on pro-inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and a pro-inflammatory cytokine, interleukin-1beta (IL-1beta) were analysed by RT-PCR, Western blot or elisa. To reveal the mechanism of action of torilin we investigated the involvement of mitogen-activated protein kinase (MAPK) cascades and their downstream transcription factors, nuclear factor-kappaB (NF-kappaB) and cyclic AMP responsive element (CRE)-binding protein (CREB). KEY RESULTS: Torilin significantly reduced the LPS-induced expression of iNOS, COX-2 and IL-1beta, and the subsequent release of NO, prostaglandin E(2) and IL-1beta into culture medium. LPS stimulation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 MAPK was inhibited by torilin. In addition, the inhibitory effect of torilin on NF-kappaB and CREB was shown by torilin-mediated recovery of LPS-induced degradation of inhibitor kappaB-alpha and suppression of LPS-induced phosphorylation of CREB respectively. CONCLUSION AND IMPLICATIONS: This study indicates that torilin inhibited LPS-induced iNOS, COX-2 and IL-1beta via down regulation of ERK1/2, p38 MAPK, NF-kappaB and CREB and suggests that torilin has a potential as an anti-inflammatory drug candidate. PMID- 19298260 TI - From assets to school outcomes: how finances shape children's perceived possibilities and intentions. AB - People do not always take action to attain their desired possible selves--after all, whether consciously or nonconsciously, taking current action makes sense if there is an open path toward attaining the desired self, but not if paths are closed. Following this logic, children from families with fewer assets may lower their expectations for school success and plan to engage in less effort in school. To test this hypothesis, we examined the impact of experimentally manipulating mind-set about college as either "closed" (expensive) or "open" (can be paid for with need-based financial aid) among low-income early adolescents. Adolescents assigned to an open-path condition expected higher grades than those assigned to a closed-path condition (Study 1, n= 48, predominantly Hispanic and Latino seventh graders) and planned to spend more time on homework than those assigned to a no-prime control condition (Study 2, n= 48, predominantly African American seventh graders). PMID- 19298261 TI - Cheating the lie detector: faking in the autobiographical Implicit Association Test. AB - The autobiographical Implicit Association Test (aIAT) was recently introduced in this journal as a new and promising lie-detection tool. The initial report found 91% accuracy in determining which of two autobiographical events was true. It was suggested that the aIAT, unlike other lie-detection tests, is resistant to faking. We investigated whether participants can strategically alter their performance on the aIAT. Experiment 1 showed that participants guilty of a mock theft were able to obtain an innocent test outcome. Two additional experiments showed that guilty participants can fake the aIAT without prior experience with the aIAT and when a response deadline is imposed. The aIAT is subject to the same shortcomings as other lie-detection tests. PMID- 19298262 TI - Living in history: how war, terrorism, and natural disaster affect the organization of autobiographical memory. AB - Memories of war, terrorism, and natural disaster play a critical role in the construction of group identity and the persistence of group conflict. Here, we argue that personal memory and knowledge of the collective past become entwined only when public events have a direct, forceful, and prolonged impact on a population. Support for this position comes from a cross-national study in which participants thought aloud as they dated mundane autobiographical events. We found that Bosnians often mentioned their civil war and that Izmit Turks made frequent reference to the 1999 earthquake in their country. In contrast, public events were rarely mentioned by Serbs, Montenegrins, Ankara Turks, Canadians, Danes, or Israelis. Surprisingly, historical references were absent from (post September 11) protocols collected in New York City and elsewhere in the United States. Taken together, these findings indicate that it is personal significance, not historical importance, that determines whether public events play a role in organizing autobiographical memory. PMID- 19298263 TI - Neural activity underlying the effect of approach-motivated positive affect on narrowed attention. AB - Positive affects high in approach motivational intensity narrow attention. The present study extended this recent finding by testing whether a neural activation associated with approach-motivation intensity--relative left frontal-central activity--would underlie the effect of appetitive stimuli on narrowed attention (as measured by local attentional bias). It also tested whether individual differences in approach motivation relate to this attentional narrowing. Results supported predictions, suggesting a common association of relative left frontal hemispheric processing for approach-motivational processes and narrowed attention. PMID- 19298264 TI - Contribution of cyclic parthenogenesis and colonization history to population structure in Daphnia. AB - Cyclic parthenogenesis, the alternation of parthenogenetic and sexual reproduction, can lead to a wide scope of population structures, ranging from almost monoclonal to genetically highly diverse populations. In addition, sexual reproduction in aquatic cyclic parthenogens is associated with the production of dormant stages, which both enhance potential gene flow among populations as well as impact local evolutionary rates through the formation of dormant egg banks. Members of the cladoceran genus Daphnia are widely distributed key organisms in freshwater habitats, which mostly exhibit this reproduction mode. We assessed patterns of genetic variation within and among populations in the eurytopic and morphologically variable species Daphnia longispina, using data from both nuclear (13 microsatellite loci) and mitochondrial (partial sequencing of the 12S rRNA gene) markers from a set of populations sampled across Europe. Most populations were characterized by very high clonal diversity, reflecting an important impact of sexual reproduction and low levels of clonal selection. Among-population genetic differentiation was very high for both nuclear and mitochondrial markers, and no strong pattern of isolation by distance was observed. We also did not observe any substantial genetic differentiation among traditionally recognized morphotypes of D. longispina. Our findings of high levels of within-population genetic variation combined with high among-population genetic differentiation are in line with predictions of the monopolization hypothesis, which suggests that in species with rapid population growth and potential for local adaptation, strong priority effects due to monopolization of resources lead to reduced levels of gene flow. PMID- 19298265 TI - Biodiversity in microbial communities: system scale patterns and mechanisms. AB - The relationship between anthropogenic impact and the maintenance of biodiversity is a fundamental question in ecology. The emphasis on the organizational level of biodiversity responsible for ecosystem processes is shifting from a species centred focus to include genotypic diversity. The relationship between biodiversity measures at these two scales remains largely unknown. By stratifying anthropogenic effects between scales of biodiversity of bacterial communities, we show a statistically significant difference in diversity based on taxonomic scale. Communities with intermediate species richness show high genotypic diversity while speciose and species-poor communities do not. We propose that in species-poor communities, generally comprising stable yet harsh conditions, physiological tolerance and competitive trade-offs limit both the number of species that occur and the loss of genotypes due to decreases in already constrained fitness. In species-rich communities, natural environmental conditions result in well-defined community structure and resource partitioning. Disturbance of these communities disrupts niche space, resulting in lower genotypic diversity despite the maintenance of species diversity. Our work provides a model to inform future research about relationships between species and genotypic biodiversity based on determining the biodiversity consequences of changing environmental context. PMID- 19298266 TI - Diversification in flowering time due to tandem FT-like gene duplication, generating novel Mendelian factors in wild and cultivated rice. AB - The complex structure of a single Mendelian factor widespread in the Asian cultivated rice (Oryza sativa) and its wild progenitor (Oryza rufipogon) that caused diverse phenotypes in the timing of flowering under natural field conditions was investigated in near isogenic lines. These near isogenic lines showed differences in flowering time despite all eight accessions collected from tropical regions possessing a recessive gene allelic to the se-pat gene. Fine mapping in two of these near-isogenic lines revealed that cultivated (Patpaku) and wild (W593) accessions had three and two linked quantitative trait loci (QTL) in the candidate regions, respectively, showing that Patpaku and W593 possessed linked QTLs with different effects in addition to the commonly-observed recessive gene (se-pat). Molecular dissection suggested that the tandemly duplicated FT like genes (Hd3a and RFT1) could be the candidate genes for these QTLs. Interestingly, the linked QTLs differed in their epistases, degree of dominance, and genotype x environment interactions. The nucleotide sequences showed that RFT1 has diverged more rapidly than Hd3a during rice evolution, suggesting phenotypic diversification of the two genes. Phylogenetic analysis implied that the se-pat(+) alleles might have emerged in different lineages within O. sativa. The present results strongly suggest that nucleotide divergence and shuffling of the linked QTLs by recombination might have created novel Mendelian factors that probably contribute to responding to local environments. PMID- 19298267 TI - Age-specific effect of heterozygosity on survival in alpine marmots, Marmota marmota. AB - The fitness consequences of heterozygosity and the mechanisms underpinning them are still highly controversial. Using capture-mark-recapture models, we investigated the effects of individual heterozygosity, measured at 16 microsatellite markers, on age-dependent survival and access to dominance in a socially monogamous mammalian species, the alpine marmot. We found a positive correlation between standardized multilocus heterozygosity and juvenile survival. However, there was no correlation between standardized multilocus heterozygosity and either survival of older individuals or access to dominance. The disappearance of a significant heterozygosity fitness correlation when individuals older than juveniles are considered is consistent with the prediction that differences in survival among individuals are maximal early in life. The lack of a correlation between heterozygosity and access to dominance may be a consequence of few homozygous individuals attaining the age at which they might reach dominance. Two hypotheses have been proposed to explain heterozygosity fitness correlations: genome-wide effects reflected by all markers or local effects of specific markers linked to genes that determine fitness. In accordance with genome-wide effects of heterozygosity, we found significant correlations between heterozygosities calculated across single locus or across two sets of eight loci. Thus, the genome-wide heterozygosity effect seems to explain the observed heterozygosity-fitness correlation in the alpine marmot. PMID- 19298268 TI - Willingness-to-pay and quality of life in patients with vitiligo. AB - BACKGROUND: Vitiligo is a chronic pigmentary disorder of the skin, affecting 1-2% of the general population. Although not life threatening, vitiligo may considerably influence patients' health-related quality of life (QoL) and psychological well-being. Willingness-to-pay (WTP) is a construct reflecting disease burden and QoL reduction which has not yet been used in vitiligo. OBJECTIVES: To assess the WTP and the QoL of patients with vitiligo. METHODS: Patients with vitiligo were included in a nationwide German postal survey. WTP was assessed by two standardized items, and QoL was evaluated using the Dermatology Life Quality Index (DLQI) and the EuroQol (EQ-5D) questionnaire. QoL data were compared with n = 1,511 patients from a national survey on psoriasis. RESULTS: The questionnaire was completed by 1,023 patients (71.5% women, mean age 44.4 years, mean disease duration 20.3 years) with vitiligo. The mean DLQI was 7.0 (7.5 in women, 5.5 in men) compared with 8.6 in psoriasis. Of the patients with vitiligo, 24.6% had a DLQI > 10 which indicates severe QoL reductions, compared with 34.1% in patients with psoriasis. The highest mean DLQI value was observed in the patient group aged 20-29 years. EQ-5D mean score was 83.6 compared with 75.3 in psoriasis. Of the patients with vitiligo, 32.9% would pay more than 5,000 Euro in order to achieve complete disease remission. WTP was highest among middle-aged patients (30-60 years). There was a significant correlation between DLQI scores and WTP (chi(2) = 65.43, P < 0.001). Moreover, WTP significantly correlated with duration of disease, and with body surface area affected (P < 0.001). CONCLUSIONS: Vitiligo causes substantial disease burden as reflected by QoL impairment and high WTP, especially in women. These results should draw the attention of physicians to this disease, as appropriate education and treatment are likely to improve the QoL of patients with vitiligo and may support patients' compliance and empowerment. PMID- 19298269 TI - Human papillomavirus-associated induction of human beta-defensins in anal intraepithelial neoplasia. AB - BACKGROUND: Antimicrobial peptides and proteins (AMPs) are widely distributed effector molecules of the innate immune system with well-known antibacterial activity. However, there is a paucity of information regarding antiviral effects of AMPs. OBJECTIVES: The present study was performed to analyse expression of AMPs in human papillomavirus (HPV)-associated anal skin lesions of human immunodeficiency virus (HIV)-positive men who have sex with men (MSM), a special high-risk group for persistent HPV infections and anal dysplasia. METHODS: Skin lesions were analysed for the presence of LL-37, RNase 7, and human beta-defensin (hBD)-1, hBD-2 and hBD-3. Moreover, HPV typing and HPV DNA load determination for HPV types 6, 11, 16, 18, 31 and 33 were performed to evaluate possible correlations between expression of AMPs and lesional HPV types. RESULTS: Skin biopsies of 45 HIV-positive MSM with anal intraepithelial neoplasia (AIN), anal condylomata acuminata or unaffected anal mucosa, as well as condylomata acuminata of eight HIV-negative MSM, were analysed for AMP mRNA expression. Additionally, immunohistochemical analysis for hBD-2 and hBD-3 was performed in a total of 45 samples. hBD-2 and hBD-3 gene and protein expression was significantly increased in both AIN and condyloma, whereas LL-37, RNase 7 and hBD-1 gene expression did not differ significantly from unaffected anal mucosa. AMP expression correlated neither with the number of HPV types nor with the high-risk and low-risk HPV DNA loads of the quantified types. No significant differences in AMP expression were observed in condylomata of HIV-positive and HIV-negative MSM. CONCLUSIONS: hBD-2 and hBD-3 expression was shown to be significantly upregulated in HPV-associated anal skin lesions of both HIV-positive and HIV-negative MSM. Their biological significance in the innate immunity against these lesions needs further research. PMID- 19298270 TI - Hyperglycaemic conditions hamper keratinocyte locomotion via sequential inhibition of distinct pathways: new insights on poor wound closure in patients with diabetes. AB - BACKGROUND: Diabetes mellitus (DM) is characterized by impaired insulin signalling, elevated plasma glucose, and predisposition towards complications involving several organs. A major complication of DM is impairment of wound healing. In the re-epithelialization process during wound healing, migration of keratinocytes is a crucial step. Our previous report demonstrated that keratinocytes cultured in hyperglycaemic media showed decreased cell mobility. OBJECTIVES: The current study aimed to explore the effects of high glucose on keratinocyte migration after different treatment durations. METHODS: Keratinocytes were cultivated for indicated time periods under various concentrations of glucose. Relevant assays including Transwell migration and in vitro wound scratch assays, flow cytometric analysis, matrix metalloproteinase-1 (MMP-1) activity assay, determination of mRNA expression and Western blotting were performed. RESULTS: We demonstrated that (i) keratinocyte motility progressively and significantly decreased; (ii) the keratinocyte activation marker K16 was significantly suppressed; (iii) expression of alpha2beta1 integrin and MMP-1, both crucial for keratinocyte locomotion on collagen type I, was significantly downregulated; and (iv) expression of the phosphorylated signal transducer and activator of transcription-1 significantly decreased after hyperglycaemic treatment. More specifically, different pathways become involved after prolonged duration of high glucose cultivation to reduce keratinocyte locomotion further. CONCLUSIONS: We have demonstrated that high glucose treatment results in progressive suppression of keratinocyte locomotion and elucidated the molecular mechanisms involved. These results provide a reasonable explanation for the poor wound healing seen in patients with DM. PMID- 19298271 TI - Depressive symptoms predict the future risk of severe pruritus in haemodialysis patients: Japan Dialysis Outcomes and Practice Patterns Study. AB - BACKGROUND: Recent reports suggest a cross-sectional association between psychiatric distress and pruritus in patients on haemodialysis (HD). However, no study has examined the likelihood of developing severe pruritus in patients on HD with depressive symptoms. OBJECTIVES: To evaluate the relationship between baseline depressive symptoms and subsequent risk of developing severe pruritus. METHODS: A longitudinal study with a 0.5-2.5-year follow-up period was performed using 1799 patients on HD who had no/mild pruritus at baseline, based on the Japan Dialysis Outcomes and Practice Patterns Study (1996-2004), a cohort study composed of a representative sample of patients on HD. We assessed pruritus after the follow-up period using a self-reported questionnaire and depressive symptoms using scores from the five-item version of the Mental Health Inventory (MHI-5). RESULTS: The 1799 patients had a mean age of 56.9 years, 59.5% were men, and 23.6% presented depressive symptoms. Multivariable analysis revealed that patients with depressive symptoms had significantly higher odds of developing severe pruritus during the 0.5-2.5-year follow-up period [adjusted odds ratio (AOR) 1.57, 95% confidence interval 1.22-2.01, P < 0.001]. In addition, a significant linear trend was observed between baseline MHI-5 scores and risk of developing severe pruritus, with AORs for third, second and first MHI-5 score quartiles of 1.08, 1.51 and 1.95, respectively (P for trend < 0.0001). CONCLUSIONS: Our results suggest that depressive symptoms measured by MHI-5 may predict the future risk of developing severe pruritus in patients on HD. PMID- 19298272 TI - Pemphigoid gestationis: early onset and blister formation are associated with adverse pregnancy outcomes. AB - BACKGROUND: It is unclear whether clinical features of pemphigoid gestationis (PG), such as timing of onset and severity, may affect pregnancy outcomes or whether the adverse outcomes in pregnancies complicated by PG are related to or worsened by systemic corticosteroid treatment. OBJECTIVES: To evaluate the associations of adverse pregnancy outcomes with clinical features, autoantibody titre of PG, and systemic corticosteroid treatment. METHODS: We conducted a retrospective cohort study recruiting 61 pregnancies complicated by PG from the St John's Institute of Dermatology database which enrolled cases from dermatologists across the U.K., and two tertiary hospitals in the U.K. and Taiwan. Outcome measures included gestational age at delivery, preterm birth, birthweight, low birthweight (LBW, i.e. birthweight < 2500 g), small-for gestational-age (i.e. birthweight below the 10th percentile for gestational age), fetal loss, congenital malformation, and mode of delivery. RESULTS: After controlling for maternal age and comorbidity, decreased gestational age at delivery was significantly associated with presence of blisters (P = 0.017) and disease onset in the second trimester (P = 0.001). Reduced birthweight was significantly associated with disease onset in the first and second trimesters (P = 0.030 and 0.018, respectively) as was also LBW [adjusted odds ratio (95% confidence interval) 13.71 (1.22-154.59) and 10.76 (1.05-110.65), respectively]. No significant associations of adverse pregnancy outcomes with autoantibody titre or systemic corticosteroid treatment were found. CONCLUSIONS: Onset of PG in the first or second trimester and presence of blisters may lead to adverse pregnancy outcomes including decreased gestational age at delivery, preterm birth, and LBW children. Such pregnancies should be considered high risk and appropriate obstetric care should be provided. Systemic corticosteroid treatment, in contrast, does not substantially affect pregnancy outcomes, and its use for PG in pregnant women is justified. PMID- 19298273 TI - A homozygous mutation in the ferrochelatase gene underlies erythropoietic protoporphyria associated with palmar keratoderma. PMID- 19298275 TI - Chronic lymphoedema complicated by septic arthritis of the hip. PMID- 19298276 TI - Safety of a new, single, integrated, closed photopheresis system in patients with cutaneous T-cell lymphoma. AB - BACKGROUND: Extracorporeal photopheresis (ECP) is an approved palliative treatment for cutaneous T-cell lymphoma (CTCL). The THERAKOS CELLEX (continuous flow separation) system (Therakos, Exton, PA, U.S.A.) has been developed from the current THERAKOS UVAR XTS system. It is designed to reduce treatment times and extracorporeal volumes, and allows the use of either a single- or a dual-needle configuration. OBJECTIVES: To assess the safety of the THERAKOS CELLEX system to provide ECP for patients with CTCL. METHODS: Patients received ECP with the THERAKOS CELLEX system for up to 6 months. The treatment schedule was defined by their current treatment and response to ECP. At least 150 treatments were required to assess safety of the new system. Safety was assessed using reports of adverse device effects (ADEs) and unanticipated ADEs (UADEs), device malfunctions and defects, vital signs, laboratory parameters and physical examinations. RESULTS: Thirteen patients were enrolled and 12 completed the study; 155 ECP treatments were initiated and 153 completed. There were no ADEs or UADEs reported during the study. The mean treatment time was shorter for patients who received dual- compared with single-needle treatments (74.4 vs. 103.0 min, P < 0.0001) and the extracorporeal volume was lower (216 vs. 266 mL). CONCLUSIONS: This new ECP system provides lower extracorporeal volumes, faster treatment times, and flexibility to use either single- or dual-needle access, while not being associated in this study with any ADEs, and therefore having a positive benefit risk ratio for patients with CTCL. PMID- 19298277 TI - Hand dermatitis and lymphoedema. AB - Hand dermatitis is common, with lymphoedema of the hand and forearm a rare complication. The mechanism of lymphoedema in such cases is poorly understood, hence management can be challenging. To investigate the underlying mechanism of lymphoedema associated with hand dermatitis and outline recommendations for management, we identified all patients with lymphoedema associated with hand dermatitis referred to our lymphoedema service, a tertiary referral centre. Treatment outcome was assessed by telephone interview and through correspondence with primary physicians and therapists. In total, nine patients, six with bilateral and three with unilateral lymphoedema associated with hand dermatitis, attended our service over a 4-year period. Most had long-standing bilateral pompholyx eczema. Three patients reported no signs of infection prior to the onset of swelling. All patients had recurrent episodes of infection after the onset of swelling. Lymphoscintigraphy, when used, revealed a failure of small initial lymphatics of the hand to absorb and drain lymph to regional nodes. Prophylactic antibiotics together with aggressive management of the dermatitis, often with systemic therapy, were required to reduce swelling. Possible mechanisms for lymphoedema associated with hand dermatitis include obliterative lymphangitis from infection, an inflammatory effect of the dermatitis on local lymphatics or a constitutive weakness of lymph drainage exposed to chronic inflammation, or any combination of the three. Treatment is only successful once both infection and inflammation from the dermatitis are controlled. PMID- 19298278 TI - Deregulation of the tumour suppressor genes p14(ARF), p15(INK4b), p16(INK4a) and p53 in basal cell carcinoma. AB - BACKGROUND: Basal cell carcinoma (BCC) is a locally aggressive slowly growing tumour that rarely metastasizes and is mostly seen in older members of the population. OBJECTIVES: To determine the involvement of the tumour suppressor genes p14(ARF), p15(INK4b), p16(INK4a) and p53 in BCC. METHODS: We investigated the integrity of the CDKN2A locus in 15 BCC samples by analysing the presence of allelic imbalance/loss of heterozygosity (LOH). Moreover, we studied the mRNA expression levels of the tumour suppressor genes p14(ARF), p15(INK4b), p16(INK4a) and p53 in the BCC samples and compared them with mRNA levels in the corresponding normal tissue. The presence of mutations was examined by sequencing for exons 1a and 2 of p16(INK4a). RESULTS: We found LOH in one BCC sample for the marker D9S1748. A polymorphism (G442A) of exon 2 was detected in three cases. p14(ARF), p15(INK4b) and p53 presented high expression levels, whereas p16(INK4a) exhibited low mRNA levels compared with the corresponding normal tissue. Significant correlations were detected among the genes studied. CONCLUSIONS: Our results demonstrate a different expression profile between p16(INK4a) and p14(ARF), p15(INK4b) and p53 in BCC. Moreover, we found a low percentage of LOH and of a polymorphic sequence variant (Ala148Thr) for the CDKN2A locus. PMID- 19298279 TI - Is there a role for insulin resistance in nonobese patients with idiopathic hirsutism? AB - BACKGROUND: Hirsutism is the presence of terminal hairs in women in a male-like pattern. It may result from various causes of androgen excess or may be idiopathic. Controversies exist concerning the presence of insulin resistance in idiopathic hirsutism (IH) or if it is a manifestation of a high body mass index (BMI). OBJECTIVES: To assess insulin resistance in nonobese patients with IH. METHODS: The study included three groups of age- and BMI-matched nonobese women: 30 patients with IH (group 1), 20 patients with hirsutism associated with polycystic ovary syndrome (PCOS) (group 2) and 20 healthy controls (group 3). The pattern of obesity based on waist to hip ratio (WHR), and insulin resistance based on fasting insulin levels and the homeostasis model assessment of insulin resistance (HOMA-IR) were assessed in all the groups. RESULTS: Sixteen patients with IH and 17 with PCOS had insulin resistance with statistically significant differences in fasting insulin levels and HOMA-IR between the three groups, between patients with IH and healthy controls and between patients with PCOS and healthy controls; there were no significant differences between patients with IH and patients with PCOS. When classified according to the pattern of obesity, 23 patients in group 1, 17 in group 2 and two in group 3 had a WHR >or= 0.85 (android obesity) with highly significant higher values of fasting insulin levels and HOMA-IR in patients with a WHR >or= 0.85 when compared with those with a WHR < 0.85. CONCLUSIONS: Insulin resistance occurs in nonobese patients with IH and appears to be related to android obesity. PMID- 19298280 TI - Objective vs. subjective factors in the psychological impact of vitiligo: the experience from a French referral centre. AB - BACKGROUND: Vitiligo affects around 0.5% of the population. The burden of disease perceived by patients seems to be underestimated by the medical community. OBJECTIVES: To analyse the impact of objective and psychological factors on the quality of life (QoL) and self-body image of patients with vitiligo. METHODS: Demographic data, medical information and psychological factors (perceived severity, trait anxiety, trait depression, trait self-esteem, body image and QoL) were prospectively collected for all patients. RESULTS: Forty-eight patients with a mean age of 43.9 years were included in the study. Vitiligo caused overall a moderate effect on patients' QoL with a 7.17 Dermatology Life Quality Index (DLQI) mean score (out of 30) without correlation with gender. According to distribution, no or minimal (DLQI 0-1), mild (2-5), moderate (6-11) and severe (12-20) impairment of QoL was found in five (10%), 14 (29%), 18 (38%) and 11 (23%) patients, respectively. The mean perceived severity evaluated by the patients was 4.91 based on visual analogue scale from 0 to 10. The self-body image was influenced by gender, perceived severity and disease characteristics. Perceived severity and patient personality were predictors of QoL impairment. Perceived severity of vitiligo was explained mainly by the patients' personality and less significantly by objective criteria. CONCLUSIONS: Subjective as well as objective factors should be included in the assessment of disease severity and follow-up of patients with vitiligo. A simple perceived severity scale is useful in clinical practice. PMID- 19298281 TI - Nestin expression in apocrine mixed tumours of the skin. PMID- 19298282 TI - Expression of hair follicle stem cells detected by cytokeratin 15 stain: implications for pathogenesis of the scarring process in cutaneous lupus erythematosus. AB - BACKGROUND: Discoid lupus erythematosus (DLE) is a scarring disease. Although the scarring and deformity may affect any part of the body, such changes have been reported to be most obvious on the face and scalp. The pathogenesis behind this scarring process is not well understood. Once lesions have scarred, recurrent disease tends to occur at the edge of the scarred lesions but not within them. OBJECTIVES: The fact that inflammation in DLE generally involves the bulge area of the follicles raises the possibility that damage to the stem cells of the bulge region may be one process leading to the permanent loss of follicles. The aim of this study was to investigate the role of the hair follicle stem cells which reside in the bulge region in the scarring process in cutaneous lupus erythematosus (CLE). METHODS: We studied the reactivity of an antibody to the CD8 antigen (C8/144B), which recognizes cytokeratin (CK) 15 and preferentially immunostains hair follicle stem cells without staining the remaining hair follicle, on skin biopsies (scalp and body lesions) from patients with CLE (36 with discoid lesions and 10 with subacute lesions). Normal scalp and body biopsy specimens served as controls. The correlation between the extent of the cytotoxic inflammatory cell infiltrate (CD8+) and the presence of stem cells was investigated. Results were analysed semiquantitatively. RESULTS: The expression of CK15 in hair follicle stem cells was variable in the DLE lesions; there was normal to moderate CK15 expression at the bulge region of hair follicles when surrounded by mild or moderate inflammatory infiltrate (CD8+), but in cases of severe inflammation, CK15 expression was weak or absent. CONCLUSIONS: The bulge region appears to be involved in this disease as part of a broader involvement of the hair follicles; it is secondarily affected by the surrounding inflammatory cell infiltrate. Expression of C8/144B diminished and was then absent, indicating either damage to stem cells or differentiation to help in the repair process. Damage to follicular stem cells may help to explain the irreversible alopecia and the scarring process which characterize this disease. PMID- 19298283 TI - Cowden syndrome: a novel mutation and overlooked glycogenic acanthosis in gingiva. PMID- 19298284 TI - Evaluation of a near-senescent human dermal fibroblast cell line and effect of amelogenin. AB - BACKGROUND: Fibroblast senescence may delay healing of chronic wounds. OBJECTIVES: To characterize a chronic human dermal fibroblast cell line (CRL 7815) with near-senescent properties, cell proliferation and production of wound healing modulating cytokines, and biosynthesis and remodelling of collagen were compared with normal human dermal fibroblasts. Also, the response of CRL-7815 fibroblasts to the extracellular matrix protein amelogenin that is beneficial in the treatment of stalled chronic wounds was studied. METHODS: Fibroblast proliferation was monitored by time-resolved growth curves and factors secreted into the culture medium containing 10% fetal bovine serum were measured by enzyme linked immunosorbent assays. Fibroblast-mediated reorganization was examined in three-dimensional type I collagen matrices. RESULTS: Cell proliferation over 9 days was significantly (P < 0.01) slower for CRL-7815 than for normal fibroblasts. Amelogenin at 1 mg mL(-1) increased (P < 0.01) CRL-7815 proliferation to the level of the normal fibroblasts. The neutrophil chemoattractant interleukin (IL)-8 was low while the constitutive production of monocyte chemoattractant protein (MCP)-1 was highly elevated in medium from cultured CRL-7815 fibroblasts. Amelogenin augmented IL-8 but attenuated MCP-1 secretion in CRL-7815 fibroblasts. The elevated vascular endothelial growth factor production in CRL-7815 fibroblasts was further increased with amelogenin while increased type I collagen synthesis by CRL-7815 was reduced with 0.1 mg mL( 1) amelogenin. The dramatically impaired collagen matrix remodelling with CRL 7815 fibroblasts (P < 0.001) was slightly improved with amelogenin (P = 0.0011). CONCLUSIONS: The near-senescent cell line CRL-7815 shares functional anomalies with fibroblasts isolated from nonhealing chronic cutaneous wounds. Amelogenin has the capacity to switch chronic fibroblasts into an acute-like phenotype. PMID- 19298285 TI - Activation of fibroblast growth factor receptor 3 and oncogene-induced senescence in skin tumours. AB - BACKGROUND: The activation of oncogenes is an important step in tumorigenesis, and recently, oncogene-induced senescence (OIS) was proposed as a critical barrier against malignant transformation in normal primary cells. OBJECTIVES: The aim of this study was to examine the activation of fibroblast growth factor receptor 3 (FGFR3) as an oncogene product and OIS in human skin tumours. METHODS: We investigated the activation of FGFR3 and OIS by mutation and immunohistochemical analysis in skin tumours, including seborrhoeic keratosis, actinic keratosis (AK), Bowen's disease (BD), basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). RESULTS: Activated point mutations of FGFR3 were identified in four of 22 cases (18%) of seborrhoeic keratosis, but no mutation was detected in the other skin tumours. Twenty-seven of 31 cases (87%) of seborrhoeic keratosis showed moderately to strongly positive expression of the FGFR3 protein, but almost all the other skin tumours were negative. On the other hand, almost all the seborrhoeic keratoses showed negative immunoreactivity for antiphoshohistone H2AX (gamma-H2AX) as a marker of OIS, but 17 of 22 cases (77%) of AK were moderately to strongly positive. Immunoreactivity for gamma-H2AX was significantly greater in AK than in seborrhoeic keratosis, BD, BCC and SCC. CONCLUSIONS: The activation of FGFR3 might be a common feature in the tumorigenesis in seborrhoeic keratosis, although the activation does not induce a typical oncogenic signal in keratinocytes. In addition, OIS due to some oncogenic signals rather than activation of FGFR3 might be involved in the early skin carcinogenesis related to chronic ultraviolet radiation exposure. PMID- 19298286 TI - Influence of the surgical technique and surface roughness on the primary stability of an implant in artificial bone with a density equivalent to maxillary bone: a laboratory study. AB - OBJECTIVE: The aim of this biomechanical study was to assess the effect of surgical technique and surface roughness on primary implant stability in low density bone. MATERIAL AND METHODS: Eighty screw-shaped (Biocomp) implants with machined or etched surface topography were inserted into a low-density bone equivalent. Solid rigid polyurethane blocks (Sawbones) with two different bone densities (group A=0.32 g/cm(3); group B=0.48 g/cm(3)) were used that are very similar to the density of the maxilla. The implant sites were prepared either by a press-fit or by an undersized technique. Peak insertion and removal torques were measured using a Digital torque gauge instrument. RESULTS: Independent of the surgical technique used, both implant types showed an increased mean insertion and removal torque value with increasing bone density. Insertion and removal torque values were 54.3+/-5.3 and 43.5+/-6.5 N cm for group A and 89.3+/ 7.6 and 55+/-9.1 for group B, respectively. For group A and B, both implant types showed statistically higher insertion and removal torque mean values for the undersized compared with the press-fit technique (P<0.01). In addition, etched implants showed statistically higher insertion and removal torque mean values compared with machined implants (P<0.01). CONCLUSION: The placement of etched implants in synthetic bone models using an undersized preparation technique resulted in enhanced primary implant stability; further, a correlation was found between primary stability and bone density of the synthetic bone, implying that in case of an implant site with low bone density, by changing the surgical technique and choosing an implant with an optimal surface roughness, the primary stability can be enhanced significantly. PMID- 19298287 TI - Biomechanical and histological behavior of zirconia implants: an experiment in the rat. AB - OBJECTIVE: This study aimed at evaluating the integration of zirconia implants in a rat femur model. MATERIAL AND METHODS: Zirconia implants with two distinct surface topographies were compared with titanium implants with similar topographies. Titanium and zirconia implants were placed into the femurs of 42 male Sprague-Dawley rats. Four groups of implants were utilized: machined zirconia implants, zirconia implants with a rough surface, machined titanium implants, and titanium implants with an electrochemically roughened surface. After a healing period of 28 days, the load-bearing capacity between the bone and the implant surface was evaluated by a push-in test. Additionally, after a healing period of 14 and 28 days, respectively, bone tissue specimens containing the implants were processed and histologically analyzed. RESULTS: The mean mineralized bone-to-implant contact showed the highest values after 14 and 28 days for the rough surfaces (titanium: 36%/45%; zirconia: 45%/59%). Also, the push-in test showed higher values for the textured implant surfaces, with no statistical significance between titanium (34 N) and zirconia (45.8 N). CONCLUSIONS: Within the limits of the animal investigation presented, it was concluded that all tested zirconia and titanium implant surfaces were biocompatible and osseoconductive. The presented surface modification of zirconia implants showed no difference regarding the histological and biomechanical results compared with an established electrochemically modified titanium implant surface. PMID- 19298288 TI - Bone repair and augmentation using block of sintered bovine-derived anorganic bone graft in cranial bone defect model. AB - OBJECTIVE: To histomorphometrically investigate the repair of critical size defects (CSDs) and bone augmentation in cranial walls using block of sintered bovine-derived anorganic bone (sBDAB) graft. MATERIAL AND METHODS: Forty guinea pigs were divided into test (n=20) and CSD control (n=20) groups. In each animal, a full-thickness bone defect with 9.5 mm diameter was made in the frontal bone. The defects were filled with an sBDAB block soaked in blood in the test group and with blood clot in the CSD control group. The skulls were collected at 0 h (n=2) and 30, 90 and 180 days (n=6/group and period) postoperatively. The volume density and total volume of newly formed bone, sBDAB, blood vessels and connective tissue, vertical thickness of removed bone plug, sBDAB block and graft area were evaluated. RESULTS: The vertical thickness of the adapted sBDAB block was 3.8 times higher than that of the removed bone plug and did not show significant difference between periods, filling in average 29.8% of the total graft region. The sBDAB block exhibited complete osseointegration with the borders of the defect at 90 days. At 90 and 180 days, the vertical thickness of the graft was 279% in the average, and the total volume of bone augmentation was, respectively, 78.8% and 148.5% higher compared with the removed bone plug. The defects of the CDS control group showed limited osteogenesis and filling by connective tissue plus tegument. CONCLUSION: The sBDAB block can be used to promote repair of CSDs and bone augmentation in the craniomaxillofacial region, due to its good osteoconductive and slow resorptive properties. PMID- 19298289 TI - Assessment of the blood supply to the lingual surface of the mandible for reduction of bleeding during implant surgery. AB - OBJECTIVES: The aim of this study was to assess the frequency of the foramina and their canals on the lingual surface of the mandible using computed tomography (CT), which was carried out for dental implant planning. MATERIAL AND METHODS: First, the visibility of the lingual canals of the CT image was verified by dissecting five cadavers. CT images of 200 patients, who had decided on implant treatment, were used in this study. The visibility of the foramina and their canals on the lingual surface of the mandible were assessed. RESULTS: The foramina were divided into two groups by the positions of the mandible, the medial lingual foramen and the lateral lingual foramen. At least one foramen was found in all patients. In the medial group, a higher level of mental spine was seen in 190 patients, the same level of mental spine was observed in 99 patients and a lower level of mental spine was observed in 114 patients. The lateral lingual foramina were found in 160/200 patients and 88/200 patients presented bilaterally. CT can predict the position and the size of the foramina and their canals on the lingual surface of the mandible. All the patients had more than one foramen in the middle of the lingual surface of the mandible on the CT image. CONCLUSION: The frequency of the lingual foramina in the medial region was 100% and that in the lateral region was 80%. It would also be useful to emphasize the significant variation in the precise location of these lingual foramina, and that these can only be visualized presurgically with volumetric imaging modalities, such as CT or Cone beam 3D systems. PMID- 19298290 TI - Mandibular single-implant overdentures: a review with surgical and prosthodontic perspectives of a novel approach. AB - OBJECTIVES: To review the literature on mandibular single-implant overdentures (opposing complete maxillary dentures), and present surgical and prosthodontic perspectives of a novel approach for this treatment option. MATERIAL AND METHODS: An electronic search through the databases of Pubmed, Embase and Medline using the linked key words 'mandibular single implant overdentures' was performed. The search was limited to English language articles published up to August 2008. Hand searches through articles retrieved from the electronic search, peer-reviewed journals and recent conference proceedings were also conducted. RESULTS: A limited number of reports were identified on mandibular single-implant overdentures (opposing maxillary complete dentures). They comprised of case series reports, short-term prospective trials and current randomized-controlled clinical trials. Different loading protocols with different implant systems have been used, but always with regular diameter implants. Specific anatomical and vascular dangers of the mandibular midline symphysis are identified including a novel surgical approach using a currently available short, wide diameter tapered implant. In addition, the prosthodontic rationale for using a larger attachment system (incorporating a platform switch) for mandibular single-implant overdentures is described. CONCLUSION: The review reveals that there is a lack of published randomized clinical trials using mandibular single-implant overdentures, opposing maxillary complete dentures. Without the evidence from randomized clinical trials, routine use of this novel approach cannot be recommended, compared with using regular diameter implants and matching attachment systems. PMID- 19298291 TI - Bone response to free form-fabricated hydroxyapatite and zirconia scaffolds: a histological study in the human maxilla. AB - OBJECTIVES: Synthetic and biological materials are increasingly used to provide temporary or permanent scaffolds for bone regeneration. This study evaluated the effect of material chemistry and microporosity on bone ingrowth and osseointegration of zirconia (ZrO(2)) and hydroxyapatite (HA) scaffolds in the human maxilla. MATERIAL AND METHODS: Twelve patients subjected to dental implant placement were enrolled in the study. Scaffolds of ZrO(2) and HA were placed in the maxilla of each subject, using a randomization protocol. After 3 months of healing, biopsies were harvested comprising the scaffolds and surrounding bone tissue. The biopsies were processed for histological evaluation and morphometric analysis (bone ingrowth and bone-to-scaffold contact). RESULTS: Healing was uneventful in all cases. All scaffolds demonstrated a measurable bone response using light microscopy and scanning electron microscopy. Microporous HA scaffolds revealed four times larger bone ingrowth and seven times larger bone contact as compared with ZrO(2) scaffolds. CONCLUSION: The results show that chemistry and microporosity of HA promote bone ingrowth and bone contact of ceramic scaffolds in human maxilla. PMID- 19298292 TI - Anatomical characteristics of the mandibular lingual foramina observed on limited cone-beam CT images. AB - OBJECTIVES: The study aimed to investigate the regional frequency and anatomical properties of mandibular lingual foramina. MATERIAL AND METHODS: The regional frequency of the lingual foramina was investigated using limited cone-beam computed tomographic images of mandibles obtained from 190 examinations of 181 patients. The mesio-distal position of the lingual foramina was classified into 17 areas of the midline area and each tooth area. The vertical position was assessed from the positional relationship between the lingual foramina and mandibular canals. Additionally, anastomoses of the lingual canals from the lingual foramina to mandibular canals were observed on the images. RESULTS: One hundred and fifty-four lingual foramina were observed. The highest regional frequency was recognized in the midline area followed by second premolar and canine area. Plural lingual foramina were observed only in the midline areas. No lingual foramina were observed in the superior area of the mandibular canals. Thirty-one lingual foramina showed anastomoses between the lingual canals and mandibular canals. CONCLUSION: Information on the high regional frequency of lingual foramina in the second premolar area would be clinically significant to avoid life-threatening incidents because of the possible presence of an artery. It is therefore important to confirm the location of lingual foramina. PMID- 19298293 TI - Effects of decontamination and implant surface characteristics on re osseointegration following treatment of peri-implantitis. AB - BACKGROUND: Although considerable bone fill may occur following treatment of peri implantitis, re-osseointegration appears to be limited and unpredictable. OBJECTIVES: To evaluate the effects of various decontamination techniques and implant surface configurations on re-osseointegration of contaminated dental implants. MATERIAL AND METHODS: Three months after tooth extraction, implants consisting of a basal part and an exchangeable intraosseous implant cylinder (EIIC) were placed in the mandibles of dogs. The EIIC was machined (M), sandblasted and acid-etched (SLA), or titanium plasma sprayed (TPS). Ligature induced peri-implantitis was initiated 8 weeks post-implantation and lasted until bone loss reached the junction of the two implant parts. Three treatment modalities were applied: (T1) the EIIC was exchanged for a pristine EIIC; (T2) the EIIC was sprayed in situ with saline; and (T3) the EIIC was removed, cleansed outside the mouth by spraying with saline, steam-sterilized, and remounted. A collagen barrier was placed over each fixture, and 3 months later, samples were processed for histology and histomorphometry. RESULTS: T2 revealed the highest bone-to-implant contact (BIC) level (significantly better than T1 and T3). T2 also yielded the highest bone crest level (significantly better than T1), followed by T3 (significantly better than T1). SLA showed the highest BIC level (significantly better than M), followed by TPS. There were no statistically significant differences in bone crest height between implant types. CONCLUSIONS: Both SLA implants and in situ cleansing resulted in the best re-osseointegration and bone fill of previously contaminated implants. PMID- 19298294 TI - Age-related efficacy of parathyroid hormone on osseointegration in the rat. AB - OBJECTIVE: Aging is a physiological determinant that can distinguish the outcome of a pharmaceutical therapy to improve osseointegration of titanium implants. Here we examined the possible interaction of intermittent parathyroid hormone (PTH) and the age of the recipient on the parameters of osseointegration in a rat tibia implantation model. MATERIAL AND METHODS: Twenty female Wistar rats aged 8 months and 20 female rats aged 2 months received PTH at 60 microg/kg body weight or a vehicle by a subcutaneous injection three times a week. After 4 weeks, histomorphometric analysis of the peri-implant area was performed. The possible interaction of the two factors on 'bone volume per tissue volume' (BV/TV) and 'bone-to-implant contact' (BIC) was tested by two-way analysis of variance. RESULTS: Based on data from the medullary compartment, two-way analysis of variance revealed that the effect of 'intermittent PTH' depends on the 'age of the recipient' when BV/TV is considered the dependent variable (P=0.04). Post hoc tests indicated that PTH leads to an increase of BV/TV of adult and young rats. However, when BIC was considered the dependent variable, no interaction was observed (P=0.14). PTH but not aging has a significant impact on BIC. In the cortical compartment, no effects of PTH on osseointegration were observed. CONCLUSIONS: The findings indicate that treatment with PTH is more effective on the peri-implant bone regeneration in adult than in young animals, supporting the importance to consider the influences of age on the development of pharmaceutical therapies to increase the process of osseointegration. PMID- 19298295 TI - Successful outcome of splinted implants supporting a 'planned' maxillary overdenture: a retrospective evaluation and comparison with fixed full dental prostheses. AB - OBJECTIVE: Whereas an overdenture concept is generally accepted for the mandible, the outcome of this therapy for the maxilla remains less documented. This retrospective analysis evaluated the peri-implant parameters of implants supporting a 'planned' overdenture in the maxilla, and compared these data with those of an age-matched control group, with a fixed full dental prosthesis. MATERIAL AND METHODS: All patients with an implant-supported overdenture in the maxilla from the Leuven University Hospital (at least 12 months in function) were recalled for a thorough examination. Forty-four patients (162 implants, Branemark type) could be enrolled. RESULTS: The mean loading time was 9 years (range: >1 to >20 years). The cumulative survival rate after 10 years of function was 99.3% if four to six interconnected implants supported the overdenture, but only 85.7% in case two non-connected implants were used as support. The marginal bone level in the former group remained stable, with changes comparable with those observed for implants supporting fixed full dental prostheses. CONCLUSIONS: These results fully support the overdenture treatment concept for the maxilla, at least when > or =4 splinted implants are used. PMID- 19298296 TI - Double-blind randomized controlled trial study on post-extraction immediately restored implants using the switching platform concept: soft tissue response. Preliminary report. AB - AIM: To evaluate the soft tissue response to immediately placed implants using the platform switching concept. MATERIAL AND METHODS: In 22 patients, 22 implants of 5.5 mm platform diameter were placed immediately into fresh extraction sockets in maxillae without compromised bone tissue. Eventual post-extraction bone defects were filled using bovine bone matrix mixed with collagen. Immediately after insertion, implants were randomly divided: 11 implants were connected with a 3.8 mm diameter abutment (test group) and 11 with a 5.5 mm diameter abutment (control group). A provisional crown was adapted and adjusted for non-functional immediate positioning. Two months later, definitive prosthetic rehabilitation was performed. Periodontal parameter, buccal peri-implant mucosal changes (REC), mesial and distal papilla height (PH) and vertical height of jumping distance (VHG) were measured at the time of implant placement, of definitive prosthesis insertion and every 6 months thereafter. RESULTS: The mean follow-up was 25 months. All implants were clinically osseointegrated. The test group showed a +0.18 mm REC gain. PH gain was +0.045 mm on average. The mean values were statistically significant (P< or =0.005) compared with the control group (PH= 0.88 mm; REC=-0.45 mm). No difference between the two groups in periodontal parameters was found. The mean value of bone filling was 7.51 mm in the test group (97.4% of VHG) and 8.57 mm in the control group (95.2% of VHG). No statistically significant difference was found between the two groups. CONCLUSIONS: This study suggests that, in a limited time period of 2 years, immediately placed implants with subsequent platform switching can provide peri implant tissue stability. PMID- 19298297 TI - Effects of implant geometry, surface properties, and TGF-beta1 on peri-implant bone response: an experimental study in goats. AB - OBJECTIVES: Despite the high success rates in implantology, the desire to use oral implants in more challenging clinical situations drives the need for continuing refinements in implant design and surface properties. In the present study, the effect of implant geometry on implant bone response was evaluated using two geometrically different implant types, i.e. screw type (St) and push-in type(Pi). Furthermore, the potential beneficial effect of an electrosprayed calcium phosphate (CaP) coating, either or not enriched with the osteoinductive growth factor TGF-beta1, on the osteogenic response was examined. MATERIAL AND METHODS: A total of 54 implants, divided into six groups (n=9), were inserted into the femoral condyles of nine goats. After an implantation period of 12 weeks, retrieved specimens were evaluated histologically and histomorphometrically. Measurements were statistically evaluated using SPSS 14.0 and analyzed using a linear regression model. RESULTS: With respect to implant design, St-implants showed an overall superior biological healing response compared with Pi-implants. Considering surface properties, the deposition of an electrosprayed CaP (2-3 microm) coating onto implants significantly increased the amount of bone-implant contact for both implant types. Additional enrichment of the CaP coating with the osteoinductive growth factor TGF-beta1 did not significantly affect peri-implant bone response. CONCLUSIONS: The results of this study indicate that a substantial improvement of the osteogenic response to titanium implants can be achieved by the deposition of an electrosprayed CaP coating. The enrichment of the coating with 1 microg TGF-beta1 has only a marginal effect. PMID- 19298302 TI - Using the posterior pelvic pain provocation test in pregnant Japanese women. AB - This study investigated the usefulness of the posterior pelvic pain provocation (PPPP) test for differentiating between pelvic pain and low back pain because of the different prognoses and treatment strategies. We recruited 263 Japanese women who were > 36 weeks' gestation and 213 were included. The PPPP test was applied to 154 women with self-reported lumbopelvic pain. The participants were divided into three groups: the PPPP test positive (PPPP+, n = 60) subgroup, PPPP test negative (PPPP-, n = 94) subgroup, and no pain group (n = 59). The disability scores of the PPPP+ subgroup were significantly higher than the scores of the PPPP- subgroup and no pain group. In the PPPP+ subgroup, the intensity of pelvic pain was significantly correlated with the disability scores, but that of low back pain was not. The PPPP test could be carried out by a midwife without side effects and could detect pregnant women with impaired ability in daily life due to lumbopelvic pain. PMID- 19298303 TI - Comparison of the clinical practice satisfaction of nursing students in Korea and the USA. AB - The objective of this research was to compare the clinical practice satisfaction of nursing students in South Korea and the United States. A self-report questionnaire on 'Satisfaction of clinical nursing practice of undergraduate students' was used to survey 131 nursing students of the Yonsei University in South Korea and 109 nursing students of the Colorado Christian University in the USA. Findings indicate that American nursing students had significantly higher clinical practice satisfaction levels than the Korean students. Two factors were identified to explain the differences. One is the different motivation for choosing a nursing major and the second is the different clinical practice environment. Although different countries utilize clinical practice differently, it is beneficial to understand the factors that influence students' satisfaction levels by comparing countries' nursing education system. PMID- 19298304 TI - Premature menopause: exploring the experience through online communication. AB - The authors explored the experience of premature menopause by analyzing 1350 posted messages by 98 women, with unique usernames, to an online discussion board over a 90 day period between December 2005 and March 2006. Four particularly problematic aspects emerged that related to the following themes: legitimizing the symptoms, the impact of diagnosis ("losing me"), treatment ("regaining me"), and feeling alone. It appeared that premature menopause, regardless of the cause, was a largely negative experience, compounded by the often lengthy process of confirming a diagnosis based on ambiguous and often vague symptoms, the scarcity of suitable information about the condition, and the subjective lack of perceived support that the women received. Furthermore, a feeling of being different, through the lack of menstruation and fertility choices, often led to relationship problems and generally a loss of well-being. Women with premature menopause would benefit from the development of tailored information and support that addresses these problematic aspects of the condition. PMID- 19298305 TI - Physicians' attitudes and practices regarding advanced end-of-life care planning for terminally ill patients at Chiang Mai University Hospital, Thailand. AB - This study examined physicians' attitudes toward advanced directives and practices for the end-of-life care at Chiang Mai University Hospital, Thailand. The data were collected from 55 physicians (24 instructors and 31 residents) using self-reported questionnaires. The majority of the participants affirmed the usefulness of the advance directive (AD) for cardiopulmonary resuscitation and respected the patients' wish for this directive, although advanced end-of-life care and resuscitation planning with the patients was limited. Mostly, the relatives were consulted regarding ADs. This study suggests that, in traditional Thai culture, physicians and families are more inclined to make decisions for the patient when they feel that it is in the patient's best interest. Further research is needed to investigate how and to what extent such attitudes can affect medical practice for end-of-life care in the context of the rapid development and consequent changes taking place in Thailand. PMID- 19298306 TI - Wealth Index association with gender issues and the reproductive health of Egyptian women. AB - This study investigated the association of the Wealth Index of married women in Egypt with a number of gender and reproductive health issues found in the 2005 Egypt Demographic Health Survey. The data from a subsample of 5249 currently married women from a total of 19,474 was examined using logistic regression analysis. The women's lowest wealth quintile predicted the intention to continue female genital cutting for their daughters, exposure to physical and sexual marital violence, not being empowered in household decisions, having a higher number of children, having an unintended last child, mothers' maltreatment of their children, the perception of a lack of health-care providers or drugs as an obstacle to receiving care, and not being covered by health insurance. The association of poverty with the aforementioned adverse health outcomes are discussed. Physicians should understand the effect of poverty on health and endeavour to influence policy-makers to reduce the poverty burden on health. PMID- 19298307 TI - Identifying patient readmission subtypes from unplanned readmissions to hospitals in Hong Kong: a cluster analysis. AB - It has been conjectured with regard to patient readmission patterns that there might be significant differences in patient characteristics, need factors, enabling resources, and health behavior. The aim of this study was to identify the profiles of readmitted patients in Hong Kong (n = 120) based on their predisposing characteristics, needs, health behavior, and enabling resources. All the readmitted patients were recruited to the study in three hospitals from 2003 to 2005. A cluster analysis yielded three clusters: Clusters 1, 2, and 3 constituted 27.5% (n = 33), 27.5% (n = 33), and 45.0% (n = 54) of the total sample, respectively. The study results show that community nurse services do affect the rate at which patients are admitted to hospital for a second time. The findings might help by providing important information that will enable health care policy-makers to identify strategies to target a specific group of patients in the hope of reducing its readmission rate. PMID- 19298308 TI - "Time for dialysis as time to live": experiences of time in everyday life of the Swedish next of kin of hemodialysis patients. AB - The aim of this study was to explore the content of time in everyday life as experienced by the next of kin of patients on hemodialysis in Sweden. Chronic renal disease often requires hemodialysis, which is a time-consuming treatment that makes it necessary to carefully plan everyday life and involves the next of kin to a large degree. This study used a descriptive design with a content analysis approach. The analysis of the data from the twenty interviews revealed the experiences of time in the everyday lives of the next of kin of a patient on hemodialysis. The content of time in everyday life can be described as follows: fragmented time, vacuous time, and uninterrupted time. The findings show how everyday life time for the next of kin is minimized and that the common life space is contracted. The next of kin must be provided with supervision in order to provide them with more of their own time in everyday life, which can benefit their health. PMID- 19298309 TI - Searching for self: the layers and labels of panic disorder: a New Zealand study. AB - The diagnosis and management of panic symptoms are relatively neglected. If not treated, people can develop a panic disorder, a condition that has an adverse effect on the client's quality of life and psychological well-being. The aim of this New Zealand research is to understand the clients' perspective of panic disorder and how it impacts on their quality of life. Ten participants were interviewed and the data were analyzed using thematic analysis. The research findings indicate that recovery from panic disorder occurs in a process of the search for self that is made up of self-understanding and the reclaiming the self. The findings provide a psychosocial perspective of panic disorder that will assist nurses who work with these clients. Nurses are pivotal in teaching clients about their body's response to stress and health anxiety, the giving of supplementary health information, and timely referral for specialist treatment. The recommendations for nurse specialist input and biopsychosocial assessments are discussed. PMID- 19298310 TI - Emotional intelligence: Part 1: the development of scales and psychometric testing. AB - This article, the first in a series of four, describes the development of two scales for deductive and inductive measurement of emotional intelligence (EI), based on the literature and the identification of the psychometric properties of the scales. The data collection comprised two parts: (i) a literature search on the subject of emotional intelligence; and (ii) psychometric testing of the scales. The Emotional Intelligence Scale, comprising 23 items, and the Emotional Reactions and Thoughts Scale, containing 25 items, were tested on a sample of 250 postnatal mothers. The response rate was 80%. An explorative factor analysis was used to investigate the construct validity of the underlying dimensions of emotional intelligence and yielded a three-factor solution for the Emotional Intelligence Scale and a four-factor solution for the Emotional Reactions and Thoughts Scale. The internal consistency of the scales was satisfactory. How well the factor solutions fit in clinical practice remains to be validated. PMID- 19298311 TI - The meaning of complementary therapy from the perspective of Thai women with breast cancer. AB - A qualitative study based on Heideggerian phenomenology was conducted with 17 Thai women who had survived breast cancer and had utilized at least one type of complementary therapy. The study explored the meaning of such therapy and the data were collected by an in-depth interview, a demographic data-recording form, and a reflective journal. The data were analyzed by using an interpretative process that was described by Cohen, Kahn, and Steeves. Six themes were generated in relation to the meaning of complementary therapy as perceived by the participants: cancer-controlling treatment; mental strengthening; mind and body therapy; self-determination; natural therapy; and conventional therapy integration. The knowledge gained from this study will help health-care providers better understand the role that complementary therapies play in the lives of women whose lives are threatened by cancer. It is important for health-care providers to be more proactive in the culturally sensitive promotion of using complementary therapies based on the women's values and preferences. PMID- 19298312 TI - Physicians' perceptions of protocol-directed weaning in an intensive care unit in Norway. AB - The aim of this paper is to identify physicians' perceptions of protocol-directed weaning from mechanical ventilation in an intensive care unit in Norway. Errors occur in the absence of procedures, plans, and a team culture. Standardized weaning protocols, or pathways, offer a systematic approach. The focus group method was used. Four themes emerged: (i) acceptance, where the participants perceived the protocol as having a positive influence on the weaning process, although some barriers were identified, such as its failure to facilitate the "difficult-to-wean" patient; (ii) indignant responses, which gave the impression that the protocol was prescribed for the benefit of the nurses; (iii) ambivalence was evident in the two different approaches to the weaning process when problems occurred, which were "directing" and "collaborating"; and (iv) continuity and professional competence were perceived as important aspects of the weaning process. An unclear pattern of responsibility and poor interprofessional collaboration and communication were reported. PMID- 19298313 TI - Assessment and measurement of health literacy: an integrative review of the literature. AB - Research linking health literacy to health knowledge, health behaviors, health outcomes, health disparity, health status, and increasing health-care costs is prevalent around the globe. Given the importance of health literacy, it is prudent to examine the tools available to assure that patients are health literate. This article provides an integrative review in order to investigate what has been developed to evaluate health literacy in the health-care setting. The research questions considered include: (i) Which instruments or screening tools are available to assess or measure health literacy in the clinical setting?; and (ii) What are the psychometric properties, advantages, and limitations of the identified tools? A number of databases are utilized to locate research specific to this topic. The research is analyzed, the findings are summarized, and the limitations are mentioned. The implications, recommendations, and the need for future research are discussed. PMID- 19298314 TI - Review of work-related stress in mainland Chinese nurses. AB - The aim of this review was to identify the specific work-related stressors of mainland Chinese nurses and to present the current state of stress research in order to discuss directions for further research. A literature search from January 2000 to June 2007 was conducted among three electronic databases. The specific stressors among Chinese nurses were effort-reward imbalance, the poor image of nursing in the community, and managerial issues. The studies were limited to either descriptive or correlation designs, a variety of stress scales were used to investigate the nurses' work-related stressors, and there were no published studies reporting stress management interventions for Chinese nurses. This review highlights that further stress research targeted to Chinese nurses needs more prospective and longitudinal studies and has to develop a consistent instrument to measure stressors. The priority issue is the initiation of stress management interventions to improve nurses' coping skills, while the long-term goal is to reduce the level of stress or eliminate the stressors through individual, organizational, and societal interventions. PMID- 19298315 TI - Application of the theory of planned behavior to oral anticoagulant therapy. AB - Anticoagulation control is imperative for individuals who are prescribed long term oral anticoagulation therapy. Therapeutic international normalized ratios decrease the risk of the thromboembolic complications that are associated with oral anticoagulation therapy. Individuals on oral anticoagulation therapy are often asked to make lifestyle modifications that can become barriers to medication adherence. The application of the theory of planned behavior to oral anticoagulation therapy can be used to assist advanced practice nurses in assessing individuals for the perceived barriers or obstacles that might interfere with the behavioral changes necessary to successfully comply with the recommended treatment plan. PMID- 19298317 TI - An association of prodynorphin polymorphisms and opioid dependence in females in a Chinese population. AB - Prodynorphin (PDYN) binds to kappa-opioid receptors and is known to regulate dopaminergic tone, making this system important for the reinforcing and rewarding properties of drugs of abuse such as opioids. The binding of dynorphins to kappa opioid receptors also produces aversive states that may affect the development of opioid dependence. Recent animal results have shown that PDYNknockout mice show decreased ethanol consumption; however, this finding was restricted to female mice. We were interested to analyse a possible gender specificity of dynorphin effects in humans and to this end three single-nucleotide polymorphisms (SNPs) in PDYN were genotyped in a Chinese population of 484 opioid dependents and 374 controls. An interaction between sex and genotype was found in female opioid dependents. Chi-squared tests for association revealed that the genotype distributions of SNPs rs1997794 (P = 0.019) and rs1022563 (P = 0.006) in the promoter and 3' region of PDYN, respectively, were found to be associated with opioid dependence. Therefore, SNPs in PDYN are significantly associated with the risk of developing opioid dependence; however, this effect may only be seen in females. These data suggest that PDYN polymorphisms should be studied in additional female opioid-dependent populations with an emphasis on the promoter and 3' regions of the gene. PMID- 19298318 TI - The effect of naltrexone on amphetamine-induced conditioned place preference and locomotor behaviour in the rat. AB - Whereas amphetamine and other psychostimulants primarily act on the dopamine system, there is also evidence that other neurotransmitter systems, such as the endogenous opioid system, modulate psychostimulant-induced effects. Several studies have investigated the role of opioid antagonists on cocaine-induced conditioned place preference (CPP), but there is limited information about the interaction with amphetamines. The aim of the present study was to investigate the effect of the opioid receptor antagonist, naltrexone (NTX) on the conditioning, expression and reinstatement of amphetamine-induced place preference. In addition, the effect of NTX on locomotor behaviour was measured during all sessions. During training, animals were conditioned with amphetamine (2 mg/kg) to induce place preference. In order to extinguish the conditioned behaviour, animals received saline for 12 days. Reinstatement of CPP was induced by a priming dose of amphetamine (0.5 mg/kg). The interaction of NTX and amphetamine was evaluated using three paradigms of CPP: with NTX (vehicle, 0.3, 1.0 and 3.0 mg/kg) administered either 30 minutes prior to amphetamine conditioning, or 30 minutes before the expression, or 30 minutes before the amphetamine priming to induce reinstatement. Naltrexone had no effect on the conditioning, the expression or the reinstatement induced by a priming dose of amphetamine. Further, NTX by itself did not induce place preference or place aversion. In contrast, NTX significantly attenuated the locomotor response to a priming dose of amphetamine without affecting general locomotor behaviour. The results suggest differences in opioid modulation of amphetamine-induced behaviours in the rat. PMID- 19298319 TI - Do genetic and individual risk factors moderate the efficacy of motivational enhancement therapy? Drinking outcomes with an emerging adult sample. AB - Research indicates that motivational enhancement therapy (MET) helps catalyze reductions in problem drinking among emerging adults. However, moderators of this intervention remain relatively unknown. Therefore, the objectives of this study were: (1) to test whether a single session of MET increased motivation to reduce drinking and drinking outcomes; and (2) to examine whether genetic dopamine D(4) receptor L (DRD4 L) and individual personality risk factors (impulsivity and novelty seeking) moderated the effects of the MET. These hypotheses were evaluated by randomly assigning a sample of emerging adult problem drinkers (n = 67) to receive a single session of MET or alcohol education. Follow-up data indicated that only individuals who were low in impulsivity, novelty seeking and/or who had the short DRD4 variable number of tandem repeats genotype evidenced differentially increased behavior change (taking steps toward reducing drinking) following the MET. PMID- 19298320 TI - Long-term haloperidol treatment (but not risperidone) enhances addiction-related behaviors in mice: role of dopamine D2 receptors. AB - The high prevalence of psychostimulant abuse observed in schizophrenic patients may be related to the development of mesolimbic dopaminergic supersensitivity (MDS) or nigrostriatal dopaminergic supersensitivity (NDS) in response to the chronic blockade of dopamine receptors produced by typical neuroleptic treatment. We compared the effects of withdrawal from long-term administration of the typical neuroleptic haloperidol (Hal) and/or the atypical agent risperidone (Ris) on MDS and NDS, behaviorally evaluated by amphetamine-induced locomotor stimulation (AILS) and apomorphine-induced stereotypy (AIS) in mice, respectively. We further evaluated the duration of MDS and investigated the specific role of dopamine D2 receptors in this phenomenon by administering the D2 agonist quinpirole (Quin) to mice withdrawn from long-term treatment with these neuroleptics. Withdrawal (48 hours) from long-term (20 days) Hal (0.5 mg/kg i.p.) (but not 0.5 mg/kg Ris i.p.) treatment potentiated both AILS and AIS. Ris co administration abolished the potentiation of AILS and AIS observed in Hal withdrawn mice. Ten days after withdrawal from long-term treatment with Hal (but not with Ris or Ris + Hal), a potentiation in AILS was still observed. Only Hal withdrawn mice presented an attenuation of locomotor inhibition produced by Quin. Our data suggest that the atypical neuroleptic Ris has a pharmacological property that counteracts the compensatory MDS and NDS developed in response to the chronic blockade of dopamine receptors imposed by Ris itself or by typical neuroleptics such as Hal. They also indicate that MDS may be long lasting and suggest that an upregulation of dopamine D2 receptors in response to long-term treatment with the typical neuroleptic is involved in this phenomenon. PMID- 19298321 TI - Alterations in pituitary-thyroid axis function among opioid-dependent subjects after acute and protracted abstinence. AB - The aim of the present study was to investigate the changes in the pituitary thyroid axis (PTA) and the time course of the hormonal alterations in subjects with opioid dependence after abstinence. Blood samples from in-patients with opioid dependence and age- and sex-matched healthy controls were collected. The severity of opioid abuse and of withdrawal symptoms was assessed. Results were compared between patients with opioid dependence (n = 30) and healthy controls (n = 30). We found that free triiodothyronine and free thyroxine levels were comparable with healthy controls while thyroid-stimulating hormone (TSH) was lower in patients in acute opioid abstinence period. Also, TSH levels in patients remained lower than controls after 30 days of abstinence. These results indicate that PTA function is altered in opioid-dependent subjects. These data highlight the importance of screening the thyroid function for individuals with chronic opioid dependence. PMID- 19298322 TI - Associations and interactions between SNPs in the alcohol metabolizing genes and alcoholism phenotypes in European Americans. AB - BACKGROUND: Alcohol dependence is a major cause of morbidity and mortality worldwide and has a strong familial component. Several linkage and association studies have identified chromosomal regions and/or genes that affect alcohol consumption, notably in genes involved in the 2-stage pathway of alcohol metabolism. METHODS: Here, we use multiple regression models to test for associations and interactions between 2 alcohol-related phenotypes and SNPs in 17 genes involved in alcohol metabolism in a sample of 1,588 European American subjects. RESULTS: The strongest evidence for association after correcting for multiple testing was between rs1229984, a nonsynonymous coding SNP in ADH1B, and DSM-IV symptom count (p = 0.0003). This SNP was also associated with maximum number of drinks in 24 hours (p = 0.0004). Each minor allele at this SNP predicts 45% fewer DSM-IV symptoms and 18% fewer max drinks. Another SNP in a splice site in ALDH1A1 (rs8187974) showed evidence for association with both phenotypes as well (p = 0.02 and 0.004, respectively), but neither association was significant after accounting for multiple testing. Minor alleles at this SNP predict greater alcohol consumption. In addition, pairwise interactions were observed between SNPs in several genes (p = 0.00002). CONCLUSIONS: We replicated the large effect of rs1229984 on alcohol behavior, and although not common (MAF = 4%), this polymorphism may be highly relevant from a public health perspective in European Americans. Another SNP, rs8187974, may also affect alcohol behavior but requires replication. Also, interactions between polymorphisms in genes involved in alcohol metabolism are likely determinants of the parameters that ultimately affect alcohol consumption. PMID- 19298323 TI - Associations of ALDH2 and ADH1B genotypes with alcohol-related phenotypes in Asian young adults. AB - BACKGROUND: Associations of ALDH2 and ADH1B genotypes with alcohol use have been evaluated largely using case-control studies, which typically focus on adult samples and dichotomous diagnostic outcomes. Relatively fewer studies have evaluated ALDH2 and ADH1B in relation to continuous drinking outcomes or at different developmental stages. This study examined additive and interactive effects of ALDH2 and ADH1B genotypes on drinking behavior in a mixed-gender sample of Asian young adults, focusing on continuous phenotypes (e.g., heavy episodic and hazardous drinking, alcohol sensitivity, drinking consequences) whose expression is expected to precede the onset of alcohol use disorders. METHODS: The sample included 182 Chinese- and Korean-American young adults ages 18 years and older (mean age = 20 years). Effects of ALDH2, ADH1B and ethnicity were estimated using generalized linear modeling. RESULTS: The ALDH2*2 allele predicted lower reported rates of alcohol use and drinking consequences as well as greater reported sensitivity to alcohol. There were significant ethnic group differences in drinking outcomes, such that Korean ethnicity predicted higher drinking rates and lower alcohol sensitivity. ADH1B status was not significantly related to drinking outcomes. CONCLUSIONS: Ethnicity and ALDH2 status, but not ADH1B status, consistently explained significant variance in alcohol consumption in this relatively young sample. Results extend previous work by showing an association of ALDH2 genotype with drinking consequences. Findings are discussed in the context of possible developmental and population differences in the influence of ALDH2 and ADH1B variations on alcohol-related phenotypes. PMID- 19298324 TI - Collateral reports in the college setting: a meta-analytic integration. AB - BACKGROUND: The majority of research examining college drinking utilizes self report data, and collateral reports have been used to verify participants' self reported alcohol use. METHODS: This meta-analytic integration examined the correspondence of over 970 collateral and participant dyads in the college setting. RESULTS AND CONCLUSIONS: Results indicated that there is little bias (mean difference) between collateral estimates of participant drinking and participant's self-report. A cumulative meta-analysis revealed that this (null) effect was stable and unlikely to be altered by subsequent research or the existence of unpublished studies. Analysis of the agreement between collaterals and participant estimates (measured by intraclass correlation coefficients; ICCs) revealed moderate levels of agreement (mean ICC = 0.501). Examination of predictors of both bias and agreement in collateral and participant reports indicates a possible intentional and protective underreporting on the part of the collaterals. Ways to reduce this bias are discussed along with the value of using collaterals to verify participant self-report in the college setting. PMID- 19298325 TI - Influence of antisocial and psychopathic traits on decision-making biases in alcoholics. AB - BACKGROUND: Although decision-making processes have become a principal target of study among addiction researchers, few studies have specifically examined decision-making among individuals with alcohol dependence (AD) and findings to date are mixed. The present study examined the relationship between AD and decision-making, and tested whether different facets of antisocial and psychopathic traits explain this association. METHODS: Participants were men with AD (n = 22), AD and comorbid antisocial personality disorder (AD + ASPD; n = 17), or a history of recreational alcohol use, but no current or lifetime symptoms of a substance use disorder, conduct disorder, or ASPD (n = 21). Decision-making was tested using the Iowa Gambling Task (IGT). RESULTS: Across groups, participants reported similar levels of awareness of the contingencies of the task, but the AD groups with and without ASPD had poorer IGT performance compared with controls (p < 0.05). A block-by-block analysis revealed that while AD had slow but steady improvement across the task, AD + ASPD exhibited initial improvement followed by a significant decrease in advantageous decision-making during the last 20 trials (p < 0.05). This was further confirmed via evidence that impulsive/antisocial personality traits but not psychopathic traits mediated poor IGT performance beyond ASPD diagnosis. CONCLUSIONS: Alcohol-dependent males favored risky choices regardless of whether they met criteria for ASPD. However, decision-making deficits were more pronounced among those with ASPD, and personality traits characterized by impulsive and antisocial tendencies mediated the relationship between AD and decision-making. PMID- 19298326 TI - Ethyl glucuronide in hair compared with traditional alcohol biomarkers--a pilot study of heavy drinkers referred to an alcohol detoxification unit. AB - BACKGROUND: Traditional biomarkers for heavy alcohol use include serum carbohydrate-deficient transferrin (CDT), the enzymes aspartate aminotransferase (AST), and alanine aminotransferase (ALT) as well as gamma-glutamyl transferase (GGT). Measurement of the nonoxidative ethanol metabolite, ethyl glucuronide (EtG) in hair, has been proposed as a new marker with superior qualities. The aim of this study was to investigate the sensitivity of EtG in hair to detect heavy alcohol use compared with CDT, AST, ALT, and GGT. We also wanted to study the quantitative relation between alcohol intake and the different biomarkers. METHODS: Sixteen patients with a history of heavy alcohol use over the previous 3 months were recruited directly after admission to a withdrawal clinic. They were thoroughly interviewed about their drinking pattern as well as relevant diseases and use of medicines or drugs. Serum was sampled and analyzed for %CDT, AST, ALT, and GGT. Hair samples were collected and analyzed for EtG. RESULTS: The mean estimated daily intake (EDI) over the previous 3 months was 206 +/- 136 g pure alcohol. All patients fulfilled the criteria for heavy alcohol use. The sensitivity to detect heavy alcohol use was 64% for %CDT, 67% for AST, 67% for ALT, 93% for GGT, and 94% for EtG. There was no correlation between the quantitative values of EDI and %CDT, AST, ALT, and GGT. There was a positive, statistically significant correlation between EDI and the level of EtG in hair. CONCLUSIONS: In this study, EtG in hair and GGT showed the best sensitivity to detect heavy alcohol use and there was a positive correlation between EDI and the concentrations of EtG in hair. Before giving recommendations for clinical practice, further studies should be carried out on larger materials and populations with a wider range of alcohol intake. PMID- 19298327 TI - Acute and chronic ethanol modulate dopamine D2-subtype receptor responses in ventral tegmental area GABA neurons. AB - BACKGROUND: Ventral tegmental area (VTA) gamma-aminobutyric acid (GABA) neurons appear to be critical substrates underlying the acute and chronic effects of ethanol on dopamine (DA) neurotransmission in the mesocorticolimbic system implicated in drug reward. VTA GABA neuron firing rate is reduced by acute ethanol and enhanced by DA via D2 receptor activation. The objective of this study was to evaluate the role of D2 receptors in acute ethanol inhibition of VTA GABA neuron activity, as well as the adaptation of D2 receptors by chronic ethanol consumption. METHODS: Using electrophysiological methods, we evaluated the effects of intraperitoneal ethanol on DA activation of VTA GABA neurons, the effects of DA antagonists on ethanol inhibition of their firing rate, as well as adaptations in firing rate following chronic ethanol consumption. Using single cell quantitative RT-polymerase chain reaction (PCR), we evaluated the expression of VTA GABA neuron D2 receptors in rats consuming ethanol versus pair-fed controls. RESULTS: In acute ethanol studies, microelectrophoretic activation of VTA GABA neurons by DA was inhibited by acute intraperitoneal ethanol, and intravenous administration of the D2 antagonist eticlopride blocked ethanol suppression of VTA GABA neuron firing rate. In chronic ethanol studies, while there were no signs of withdrawal at 24 hours, or significant adaptation in firing rate or response to acute ethanol, there was a significant down-regulation in the expression of D2 receptors in ethanol-consuming rats versus pair-fed controls. CONCLUSIONS: Inhibition of DA activation of VTA GABA neuron firing rate by ethanol, as well as eticlopride block of ethanol inhibition of VTA GABA neuron firing rate, suggests an interaction between ethanol and DA neurotransmission via D2 receptors, perhaps via enhanced DA release in the VTA subsequent to ethanol inhibition of GABA neurons. Down-regulation of VTA GABA neuron D2 receptors by chronic ethanol might result from persistent DA release onto GABA neurons. PMID- 19298328 TI - Association between personality traits and ALDH2 polymorphism in Japanese male alcoholics. AB - BACKGROUND: Alcoholics who have developed alcoholism despite a strong negative risk factor, that is, the inactive form of aldehyde dehydrogenase-2 (ALDH2), are considered advantageous for studying predisposing factors for alcoholism. This study aimed to compare personality profiles and clinical characteristics between alcoholics with active and inactive ALDH2. METHODS: Subjects were 460 male Japanese alcoholics hospitalized in Kurihama Alcoholism Center. All patients underwent Cloninger's Tridimensional Personality Questionnaire and semi structured interviews 4 to 8 weeks after admission to obtain data on personalities and clinical characteristics. ALDH2 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Sixty-six patients had the inactive form of ALDH2 (ALDH2*1/2*2) and 394 had the active form (ALDH2*1/2*1). RESULTS: Alcoholics with inactive ALDH2 had significantly higher novelty-seeking (NS) and lower harm-avoidance (HA) scores compared with those with active ALDH2. The inactive ALDH2 group experienced delirium tremens significantly less frequently than the active ALDH2 group. CONCLUSIONS: These results suggest that high NS and low HA scores in alcoholics with inactive ALDH2 are associated with an increased risk for developing alcoholism, despite a low enzymatic ability to eliminate toxic acetaldehyde in these subjects. A study of alcoholics with inactive ALDH2 is useful for detecting environmental or personality factors related to alcoholism. PMID- 19298329 TI - Ethanol blocks adenosine uptake via inhibiting the nucleoside transport system in bronchial epithelial cells. AB - BACKGROUND: Adenosine uptake into cells by nucleoside transporters plays a significant role in governing extracellular adenosine concentration. Extracellular adenosine is an important signaling molecule that modulates many cellular functions via 4 G-protein-coupled receptor subtypes (A(1), A(2A), A(2B), and A(3)). Previously, we demonstrated that adenosine is critical in maintaining airway homeostasis and airway repair and that airway host defenses are impaired by alcohol. Taken together, we hypothesized that ethanol impairs adenosine uptake via the nucleoside transport system. METHODS: To examine ethanol-induced alteration on adenosine transport, we used a human bronchial epithelial cell line (BEAS-2B). Cells were preincubated for 10 minutes in the presence and absence of varying concentrations of ethanol (EtOH). In addition, some cells were pretreated with S-(4-Nitrobenzyl)-6-thioinosine (100 microM: NBT), a potent adenosine uptake inhibitor. Uptake was then determined by addition of [(3)H]-adenosine at various time intervals. RESULTS: Increasing EtOH concentrations resulted in increasing inhibition of adenosine uptake when measured at 1 minute. Cells pretreated with NBT effectively blocked adenosine uptake. In addition, short-term EtOH revealed increased extracellular adenosine concentration. Conversely, adenosine transport became desensitized in cells exposed to EtOH (100 mM) for 24 hours. To determine the mechanism of EtOH-induced desensitization of adenosine transport, cAMP activity was assessed in response to EtOH. Short-term EtOH exposure (10 minutes) had little or no effect on adenosine-mediated cAMP activation, whereas long-term EtOH exposure (24 hours) blocked adenosine-mediated cAMP activation. Western blot analysis of lysates from unstimulated BEAS-2B cells detected a single 55 kDa band indicating the presence of hENT1 and hENT2, respectively. Real-time RT-PCR of RNA from BEAS-2B revealed transcriptional expression of ENT1 and ENT2. CONCLUSIONS: Collectively, these data reveal that acute exposure of cells to EtOH inhibits adenosine uptake via a nucleoside transporter, and chronic exposure of cells to EtOH desensitizes the adenosine transporter to these inhibitory effects of ethanol. Furthermore, our data suggest that inhibition of adenosine uptake by EtOH leads to an increased extracellular adenosine accumulation, influencing the effect of adenosine at the epithelial cell surface, which may alter airway homeostasis. PMID- 19298330 TI - Age of drinking onset and injuries, motor vehicle crashes, and physical fights after drinking and when not drinking. AB - BACKGROUND: Earlier age of drinking onset has been associated with greater odds of involvement in motor vehicle crashes, unintentional injuries, and physical fights after drinking. This study explores whether early drinkers take more risks even when sober by comparing potential associations between age of drinking onset and these outcomes after drinking relative to when respondents have not been drinking. METHOD: From a national sample, 4,021 ever-drinkers ages 18 to 39 were asked age of drinking onset, not counting tastes or sips. They were also asked if they were ever in motor vehicle crashes, unintentionally injured, or in physical fights after drinking and when not drinking. GEE logistic regression models for repeated measures dichotomous outcomes compared whether odds ratios between age of onset and these adverse outcomes significantly differed when they occurred after drinking versus when not drinking, controlling for respondents' demographic characteristics, cigarette and marijuana use, family history of alcoholism, ever experiencing alcohol dependence, and frequency of binge drinking. RESULTS: Compared with persons who started drinking at age 21+, those who started at ages <14, 14 to 15, 16 to 17, and 18 to 20 had, after drinking, respectively greater odds: 6.3 (2.6, 15.3), 5.2 (2.2, 12.3), 3.3 (1.5, 7.3), and 2.2 (0.9, 5.1) of having been in a motor vehicle crash; 6.0 (3.4, 10.5), 4.9 (3.0, 8.6), 3.7 (2.4, 5.6), and 1.9 (1.2, 2.9) of ever being in a fight; and 4.6 (2.4, 8.7), 4.7 (2.6, 8.6), 3.2 (1.9, 5.6), and 2.3 (1.3, 4.0) of ever being accidentally injured. The odds of experiencing motor vehicle accidents or injuries when not drinking were not significantly elevated among early onset drinkers. The odds of earlier onset drinkers being in fights were also significantly greater when respondents had been drinking than not drinking. CONCLUSION: Starting to drink at an earlier age is associated with greater odds of experiencing motor vehicle crash involvement, unintentional injuries, and physical fights when respondents were drinking, but less so when respondents had not been drinking. These findings reinforce the need for programs and policies to delay drinking onset. PMID- 19298331 TI - Effects of D-cycloserine on extinction and reconditioning of ethanol-seeking behavior in mice. AB - BACKGROUND: D-cycloserine (DCS), a partial N-methyl-D-aspartate receptor agonist, has been shown to enhance the extinction of both cocaine and amphetamine-induced conditioned place preference (CPP). However, there have been no reports of the effects of DCS on the extinction of ethanol-conditioned behaviors in mice. Thus, the current experiments examined the effects of DCS on the extinction and subsequent reconditioning of ethanol-induced CPP in mice. METHODS: Male DBA/2J mice received either 2 or 4 pairings of ethanol (2 g/kg) with a conditioned stimulus (CS+) floor cue (and an equal number of saline pairings with a CS- floor cue on alternate days) resulting in either a weak or strong ethanol CPP, respectively. Following conditioning of a strong ethanol CPP mice received saline or 30 mg/kg DCS prior to each of the twelve 30-minute choice extinction trials administered at 48-hour intervals. Mice that had received conditioning of a weak ethanol CPP received saline, 30 or 60 mg/kg DCS immediately before each of the six 30-minute choice extinction trials. Following successful ethanol CPP extinction, mice received reconditioning trials similar to the initial conditioning trials. A final experiment examined the effects 12 DCS pre-exposures (15, 30, and 60 mg/kg) on initial conditioning of ethanol CPP. RESULTS: First, we showed that 2 doses of DCS (30 and 60 mg/kg) did not have aversive properties that could confound the effects on extinction of CPP (Experiment 1). Second, we showed that DCS (30 and 60 mg/kg) had no effect on the rate of extinction of either strong (Experiment 2) or weak (Experiment 3) ethanol-induced CPP. Interestingly, DCS administered during extinction interfered with reconditioning of ethanol-induced CPP--an effect specific to reconditioning, as DCS pre-exposure did not influence initial ethanol CPP conditioning (Experiment 4). CONCLUSIONS: These experiments show that although DCS showed no effect on extinction behavior, when given during extinction it interfered with subsequent reconditioning of ethanol CPP. The mechanisms of this effect were not, however, due to nonspecific interference with learning because repeated DCS pre-exposures did not impair initial conditioning of ethanol CPP. PMID- 19298332 TI - Influence of a drinking quantity and frequency measure on the prevalence and demographic correlates of DSM-IV alcohol dependence. AB - BACKGROUND: Recent research suggests that adding a quantity/frequency alcohol consumption measure to diagnoses of alcohol use disorders may improve construct validity of the diagnoses for Diagnostic and Statistical Manual of Mental and Behavior Disorders (DSM-V). This study explores the epidemiological impact of including weekly at-risk drinking (WAD) in the DMS-IV diagnostic definition of alcohol dependence via 3 hypothetical reformulations of the current criteria. METHODS: The sample was the National Epidemiologic Survey on Alcohol and Related Conditions, a nationally representative sample with 43,093 adults aged >18 in the U.S interviewed with the Alcohol Use Disorder and Associated Disabilities Interview Schedule IV. The current (DSM-IV) definition of alcohol dependence was compared with 4 hypothetical alcohol dependence reformulations that included WAD: (1) WAD added as an eighth criteria; (2) WAD required for a diagnosis; (3) adding abuse and dependence criteria together, and including WAD with a 3 of 12 symptom threshold; (4) adding abuse and dependence criteria together, and including WAD with a 5 of 12 symptom threshold. RESULTS: The inclusion of at-risk drinking as an eighth criterion of alcohol dependence has a minimal impact on the sociodemographic correlates of alcohol dependence but substantially increases the prevalence of dependence (from 3.8% to 5.0%). At-risk drinking as a required criterion or as part of a diagnosis that combines abuse with dependence criteria with a higher threshold (5+ criteria) decreases prevalence and has a larger impact on sociodemographic correlates. Blacks, Hispanics, and women are less likely to be included in diagnostic reformulations that include WAD, whereas individuals with low-income and education are more likely to remain diagnosed. CONCLUSIONS: Including WAD as either a requirement of diagnosis or as an additional criterion would have a large impact on the prevalence of alcohol dependence in the general population. The inclusion of a quantity/frequency requirement may eliminate false positives from studies of alcohol disorder etiology and improve phenotype definition for genetic association studies by reducing heterogeneity in the diagnosis, but may also reduce eligibility for treatment services among women and racial/ethnic minorities compared. PMID- 19298333 TI - Stimulatory effect of N-methyltyramine, a congener of beer, on pancreatic secretion in conscious rats. AB - BACKGROUND: Alcoholic beverages stimulate gastric acid secretion and increase the appetite. Although ingested ethanol stimulates pancreatic secretion, alcoholic beverages contain several congeners. N-methyltyramine (NMT) was isolated from beer as a factor in stimulating gastric acid secretion. In this study, we examined NMT to determine whether the congener stimulated pancreatic secretion in conscious rats. METHODS: Cannulae were inserted into male Wistar rats to separately drain bile and pancreatic secretions: 2 duodenal cannulae, a gastric cannula, and an external jugular vein cannula. The rats were placed in modified Bollman-type restraint cages. After a 4-day recovery period, experiments were conducted on unanesthetized rats. Different concentrations of NMT (5, 25, and 50 microg/kg) solutions were infused into the stomach. To examine the mechanism, the effects of the proton pump inhibitor, cholecystokinin (CCK-BR) antagonist (YM022), CCK-AR antagonist (CR1505), and atropine were administered prior to the NMT (25 microg/kg) infusion. The effect of intravenous infusion of NMT (7.5 microg/kg) was then determined. Moreover, dispersed acini were prepared, and the effect of different concentrations of NMT on amylase release was determined. RESULTS: Intragastric administration of NMT significantly increased pancreatic exocrine secretion in a dose-dependent manner. Atropine eliminated the stimulatory effect of NMT, but the infusion of the proton pump inhibitor, YM022, and CR1505 did not. Intravenous infusion of NMT did not affect pancreatic secretion, and NMT did not stimulate amylase release in vitro. CONCLUSIONS: N methyltyramine stimulates pancreatic secretion via the cholinergic gastro pancreatic reflex. The NMT content in beer was 2 mg/l, so that if a person weighing 60 kg consumes a 750 ml of beer, 25 microg/kg NMT will be ingested. Therefore, the stimulatory effect of beer on pancreatic secretion was produced not only by ethanol but also by the congener, NMT. PMID- 19298334 TI - High-risk population for gastric cancer development based on serum pepsinogen status and lifestyle factors. AB - BACKGROUND: Gastric atrophy is a major risk factor for non-cardiac gastric cancer. Serum pepsinogen status could identify people at high-risk for gastric cancer development during our previous cohort study. However, lifestyle-related factors may additionally affect this risk. MATERIALS AND METHODS: A total of 6983 Japanese were followed up by annual endoscopy in the previous study, and 43 cases of gastric cancer including two cardiac cancers developed. In most subjects, the body length and weight were measured and a questionnaire was applied to gather information regarding life habits. The risk of non-cardiac gastric cancer development during surveillance was re-analyzed based on serum pepsinogen, sex, age, body mass index (BMI), alcohol, and smoking habit. RESULTS: A total of 6158 subjects with 37 non-cardiac gastric cancer development (male/female = 4259/1899, mean age = 49.0, mean follow-up period = 4.79 years) were entered into analysis. In a multivariate analysis, old age (by 10 years; (odds ratio) OR, 2.8; p < .001), alcohol (weekly; OR, 2.4; p = .03), smoking (current; OR, 5.6; p = .006 and past; OR, 3.9; p = .04), and pepsinogen status ("atrophic"; OR, 6.2; p < .001) were independent risk factors, whereas BMI was not. The annual incidence of gastric cancer was 1.2% in the older subjects aged > or = 60 years with "atrophic" pepsinogen status. Moreover, it was as high as 2.9% when they had both alcohol and current smoking habits. CONCLUSIONS: Old age, alcohol, and smoking habits additionally promoted the risk for gastric cancer in subjects with gastric atrophy. PMID- 19298335 TI - Comparison of a stool antigen test and serology for the diagnosis of Helicobacter pylori infection in mass survey. AB - BACKGROUND: Serum antibody to Helicobacter pylori is tested in mass screening for gastric cancer along with the level of serum pepsinogens (PG) I and II. Recently, stool antigen tests have been developed as a new non-invasive test. We examined H. pylori infection by both serology and stool antigen test in a mass survey and compared the results to estimate applicability of stool antigen test for mass survey. METHODS: A total of 994 healthy adults who received mass survey in April 2005 were tested. There were 379 men and 615 women, and the mean age was 57.7 years old. Stool samples were used to measure a H. pylori-specific antigen by enzyme immunoassay. Serum samples were tested for the prevalence of IgG antibody to H. pylori, and the level of PGs I and II was also measured to determine the presence of atrophic gastritis. RESULTS: Infection of H. pylori was defined as positive 61.4% and 56.4% by serology and stool antigen test, respectively. The concordance of both tests was not affected by gender and age of the subjects but difference was seen in subjects with atrophic gastritis. In particular, positivity of stool antigen test (81.8%) was significantly lower than that of serology (88.7%, p < .05) in 303 subjects with severe atrophic gastritis. CONCLUSIONS: Stool antigen test, which detects present but not previous infection of H. pylori, would be applicable to diagnose H. pylori infection in mass survey. Usefulness of stool antigen tests for the screening of gastric cancer should be examined. PMID- 19298336 TI - Impact of quadruple regimen of clarithromycin added to metronidazole-containing triple therapy against Helicobacter pylori infection following clarithromycin containing triple-therapy failure. AB - BACKGROUND: The establishment of an optimal second-line regimen for Helicobacter pylori infection is required. Although quadruple therapy should overcome resistance to either clarithromycin or metronidazole, the effects of a quadruple regimen in second-line therapy are unknown. This study aims to evaluate the efficacy of triple therapy composed of proton pump inhibitor/amoxicillin plus metronidazole with the combined additive effects of clarithromycin as a second line quadruple therapy against H. pylori infection. MATERIALS AND METHODS: Participants were 104 patients in whom first-line therapy containing proton pump inhibitor-amoxicillin-clarithromycin failed. Before starting second-line therapy, patients underwent endoscopy to obtain H. pylori strain for antibiotic susceptibility tests. Patients were randomized to receive rabeprazole (10 mg), amoxicillin (750 mg), and metronidazole (250 mg), either with clarithromycin (200 mg; RAMC group) or without (RAM group); all treatments were administered twice daily for 7 days. H. pylori eradication was confirmed by (13)C-urea breath tests performed 2 to 3 months post-therapy. RESULTS: As shown by intention-to-treat/per protocol analyses, the cure rates for H. pylori infection were 88.5%/93.9% and 82.7%/84.3% for the RAMC and RAM groups. Although the study probably had an insufficient power to show a significant difference between the cure rates of the two regimens, the eradication rates showed a clear trend in favor of the RAMC group. There were no severe side-effects in any group. CONCLUSIONS: In Japan, the RAMC regimen is thought to be a promising alternative strategy for second-line eradication of H. pylori infection. PMID- 19298337 TI - A randomized clinical trial to determine the efficacy of regimens containing clarithromycin, metronidazole, and amoxicillin among histologic subgroups for Helicobacter pylori eradication in a developing country. AB - BACKGROUND: Most treatments deemed effective for Helicobacter pylori eradication in developed countries are less effective in developing countries. Regimens containing clarithromycin, metronidazole, and amoxicillin seem efficacious despite antibiotic resistance, and may be a viable option in developing countries. MATERIALS AND METHODS: We evaluated the efficacy of a 14-day regimen with 500 mg clarithromycin b.i.d., 500 mg metronidazole t.i.d., and 500 mg amoxicillin t.i.d. (with and without a proton pump inhibitor), and a 10-day regimen containing 500 mg clarithromycin b.i.d., 1 g amoxicillin b.i.d., and 20 mg omeprazole b.i.d. in Pasto, Colombia, using a randomized, single-blind design stratified by presence of atrophic gastritis. RESULTS: H. pylori was eradicated in 86.8% and 85.3% of the participants randomized to a clarithromycin metronidazole-amoxicillin and clarithromycin-amoxicillin-omeprazole regimens, respectively (p = .79). Per-protocol analyses indicated greater efficacy for the clarithromycin-metronidazole-amoxicillin regimen (97%) versus the clarithromycin amoxicillin-omeprazole regimen (86%) (p = .04), particularly for participants with atrophic gastritis (clarithromycin-metronidazole-amoxicillin = 100%, clarithromycin-amoxicillin-omeprazole = 81%; p = .02). Adverse events were mild, but adverse event-related non-compliance was reported more often for regimens containing clarithromycin, metronidazole, and amoxicillin. CONCLUSIONS: Our results suggest that an eradication rate of > 85% can be achieved with 14-day clarithromycin, metronidazole, and amoxicillin and 10-day clarithromycin, amoxicillin, and omeprazole regimens in Pasto, Colombia. The regimens containing clarithromycin, metronidazole, and amoxicillin appear to be superior to the clarithromycin, amoxicillin, and omeprazole regimen for compliant participants and those with atrophic gastritis. Our findings provide treatment options for a population in a developing country with a high prevalence of H. pylori infections and antibiotic resistance. PMID- 19298338 TI - Meta-analysis: four-drug, three-antibiotic, non-bismuth-containing "concomitant therapy" versus triple therapy for Helicobacter pylori eradication. AB - BACKGROUND: Low success rates with triple therapy for Helicobacter pylori infections have prompted search for alternatives. In one, a proton-pump inhibitor (PPI) and amoxicillin was followed by the PPI plus clarithromycin and a nitroimidazole (sequential therapy); in another, these four drugs were given concomitantly (concomitant therapy). AIM: To compare concomitant therapy with standard triple therapy for H. pylori infection. METHODS: By searching PubMed, EMBASE, the Cochrane Central Register of Controlled Trials and abstracts of major gastrointestinal meeting, two independent reviewers systemically identified randomized controlled trials (RCT) comparing concomitant quadruple to standard triple therapies as well as studies reporting eradication rates of concomitant quadruple therapy in treatment of H. pylori. Pooled eradication rates and odds ratios (OR) with 95% confidence intervals (CI) were calculated, and univariable metaregression analysis for all extracted variables was conducted. RESULTS: We identified nine studies (10 treatment arms) including five qualifying RCTs (576 subjects) comparing concomitant (293 subjects, duration 3 to 5 days) and triple therapy (283 subjects, duration 5 to 10 days) and four other studies evaluating concomitant therapy (478 subjects, duration 3 to 7 days). Pooled estimates of the five RCTs showed superiority of concomitant therapy over triple therapy; with intention-to-treat) pooled OR of 2.86 (95% CI: 1.73-4.73) and per-protocol (PP) pooled OR of 3.52 (95% CI: 1.95-6.38). Considering all 10 treatment arms, the ITT eradication rate was 89.7% (95% CI: 86.8-92.1%) and PP was 92.9% (95% CI: 90.2 94.8%). CONCLUSION: Concomitant therapy appears to be an effective alternative to triple therapy and is less complex than sequential therapy. PMID- 19298339 TI - Meta-analysis: the effect of supplementation with lactoferrin on eradication rates and adverse events during Helicobacter pylori eradication therapy. AB - BACKGROUND: Recent evidence shown that lactoferrin could exert an antimicrobial effect against Helicobacter pylori both in vitro and in vivo models. To systematically evaluate whether adding lactoferrin to H. pylori eradication regimens could improve eradication rates and reduce side-effects during anti-H. pylori treatment. MATERIALS AND METHODS: Eligible articles were identified by searches of electronic databases. We included all randomized trials comparing lactoferrin supplementation to placebo or no treatment during anti-H. pylori regimens. Statistical analysis was performed with Review Manager 5.0.10. Subanalysis/Sensitivity analysis was also performed. RESULTS: We identified nine randomized trials (n = 1343). Pooled H. pylori eradication rates were 86.57% (95% confidence interval (CI) = 83.99-89.15%) and 74.44% (95% CI = 71.14-77.74%) for patients with or without lactoferrin by intention-to-treat analysis, respectively, the odds ratio (OR) was 2.26 (95% CI = 1.70-3.00); the occurrence of total side-effects was 9.05% (95% CI = 6.83-11.27%) and 16.28% (95% CI = 13.43%-19.13%) for groups with or without lactoferrin, especially for nausea, the summary OR was 0.15 (95% CI = 0.04-0.54). CONCLUSIONS: Our review suggests that supplementation with lactoferrin could be effective in increasing eradication rates of anti-H. pylori therapy, and could be considered helpful for patients with eradication failure. Furthermore, lactoferrin shows a positive impact on H. pylori therapy-related side-effects. PMID- 19298340 TI - Prevalence of Helicobacter pylori infection and identification of risk factors in rural and urban Beijing, China. AB - BACKGROUND: The prevalence of Helicobacter pylori is higher in developing countries such as China. The aim of this study was to investigate the prevalence of H. pylori in one rural and one urban region of Beijing, China. MATERIALS AND METHODS: Healthy individuals in rural Pinggu and urban Haidian voluntarily participated in this study. The diagnosis of H. pylori infection was reached using the (13)C-urea breath test. Associations between H. pylori and sex, age, living area (i.e. rural vs urban), education level, smoking, and alcohol consumption were evaluated. RESULTS: Of the 1232 included subjects, 54.7% of tested individuals residing in Pinggu and 41.3% in Haidian were positive for H. pylori. In urban region, more individuals were negative for H. pylori (429 of 731), whereas in the rural region, more individuals were positive for H. pylori (p < .05). Univariate analysis identified geographic area and lower education and annual income as significant factors associated with H. pylori. Men in rural areas were more likely than women in rural areas to be infected, and both men and women in the rural area were more likely to be positive for H. pylori than men and women in the urban area (all p < .05). CONCLUSIONS: H. pylori infection is common in both rural and urban regions of Beijing. Residing in a rural area, having a lower family income, and lower education level are significant risk factors associated with infection. PMID- 19298341 TI - Urea breath test in children: the United States prospective, multicenter study. AB - BACKGROUND: The urea breath test (UBT) is generally considered the gold standard for the diagnosis of Helicobacter pylori infections in adults. GOALS: To investigate the utility and accuracy of urea breath testing in children from the United States. METHODS: Children scheduled to undergo upper gastrointestinal endoscopy for various clinical symptoms underwent a 13C-UBT using the US standard protocol for adults. Results were compared with rapid urease testing (RUT), culture, and histology. H. pylori positivity was defined according to the FDA, Division of Anti-Infective Drug Products criteria, i.e. positive culture and/or positive RUT and histology. H. pylori negativity was defined as all tests negative. Results were evaluated by delta over baseline (DOB) and urea hydrolysis rate (UHR). RESULTS: A total of 176 children from five centers were evaluated; 48 were infected. Compared to the defined standard, the results with the UBT based on delta over baseline (DOB) cut-off value (positive: > or = 2.4 per thousand) showed that the sensitivity and specificity of the UBT were 97.9% and 96.1%, respectively. Based on the UHR cut-off value (positive: > or = 10.0 microg/min), the sensitivity and specificity were 95.8% and 99.2%. In young children (2- to 5 year olds), sensitivity and specificity of UHR method were higher than the DOB method (100% and 100% vs 100% and 82.4%, respectively). CONCLUSION: The US standard (13)C-UBT proved to be both simple and accurate for the diagnosis of H. pylori infections in children. The UHR method to calculate of (13)C-UBT result provided excellent results for children of all ages. PMID- 19298342 TI - Fur-independent induction of Helicobacter pylori flavodoxin-encoding gene (fldA) under iron starvation. AB - BACKGROUND AND AIMS: Helicobacter pylori infection is associated with a variety of diseases including gastric cancer. Flavodoxin is an electron transfer protein containing a flavin mononucleotide prosthetic group and substituted an iron containing electron transfer protein under iron-limiting conditions. H. pylori flavodoxin has been reported but its pathogenic role is unclear. The aim of this study is to understand a pathogenic role of H. pylori flavodoxin under iron limiting condition. METHODS: The flavodoxin-encoding gene (fldA) was cloned from one of clinical H. pylori isolates (DU17) and its transcript was quantified by primer extension, Northern hybridization, and real-time polymerase chain reaction in different concentrations of an iron chelator. The fldA transcript was also quantified in H. pylori ATCC 700392, lacking a ferric uptake regulatory (fur) protein. RESULT: Nucleotide sequence of the fldA from H. pylori DU17 revealed a 492-bp (164 amino acids) open reading frame with a deduced amino acid sequence having 97% identity to that from the complete genomic sequence of H. pylori 26695. The deduced promoter [-35, -10, and +1] of the fldA was 56-bp upstream from the first codon of FldA. The fldA transcript (approximately 0.55-kb) was induced up to 14-fold in both wild-type and fur-knocked-out strains under iron limiting conditions, suggesting that the fldA induction is independent to the Fur protein. CONCLUSION: The fldA gene may play an important role in iron starvation conditions. PMID- 19298343 TI - Presence of thrombin-activatable fibrinolysis inhibitor in Helicobacter pylori associated gastroduodenal disease. AB - Thrombin-activatable fibrinolysis inhibitor (TAFI) plays a role in the regulation of coagulation and inflammation. In addition to inhibiting the fibrinolytic system, TAFI may also regulate the bradykinin and complement systems. We hypothesized that TAFI also plays a role in defense mechanisms of the gastric mucosa during Helicobacter pylori infection. This study comprised 65 patients with gastroduodenal disorders: 41 patients with H. pylori infection, 13 without, and 11 patients with cured H. pylori infection. The gastric intramucosal concentrations of TAFI were measured by enzyme immunoassay. The gastric levels of TAFI and plasminogen activator inhibitor-1 were significantly increased in patients with H. pylori compared to those without infection or cured H. pylori. The presence of TAFI was detected in gastric mucosal epithelial cells. The concentration of TAFI was correlated with the degree of gastric mucosal atrophy, inflammation, and disease activity. These results show that TAFI is present in the gastric mucosa and that it may play a role in the pathogenesis of H. pylori infection-associated gastroduodenal disorders. PMID- 19298344 TI - Evaluation of clarithromycin resistance in Helicobacter pylori obtained from culture isolates, gastric juice, and feces. PMID- 19298345 TI - How to write in the active voice. PMID- 19298346 TI - Molecular markers for detection, surveillance and prognostication of bladder cancer. AB - Many markers for the detection of bladder cancers have been tested and almost all urinary markers reported are better than cytology with regard to sensitivity, but they score lower in specificity. Currently molecular and genetic changes play an important role in the discovery of new molecular markers for detection, prognostication and surveillance. The purpose of this review is to highlight the most important urinary molecular biomarker developments that have been studied and reported recently. In the current review we have summarized the most recent and relevant published reports on molecular urinary markers. The results of this review show that the first generation of urinary markers did not add much to urinary cytology. The current generation of markers is better, but additional clinical trials are needed. Our knowledge of molecular pathways in bladder cancer is growing and new methods of marker development emerge, but the perfect marker is still to be found. Currently, there are not clinically usable molecular markers that can guide us in diagnosis or surveillance, nor guide us in lowering the frequency of urethrocystoscopy in bladder cancer. PMID- 19298350 TI - An aggressive course of Xp11 translocation renal cell carcinoma in a 28-year-old man. AB - Cases of renal cell carcinoma (RCC) associated with Xp11 translocations are rare and are reported predominantly in children. We report a case of a young man who developed an aggressive Xp11 translocation RCC. A 28-year-old man presented with back pain, fever and macroscopic hematuria. Computed tomography of the abdomen showed a heterogeneous mass in the left kidney. Left radical nephrectomy was performed. Hematoxylin-eosin staining revealed nested and papillary architecture, clear and eosinophilic cytoplasm and vesicles with prominent nucleoli. Immunohistochemical evaluation revealed that the tumor cells showed nuclear labeling for TFE3 protein. On the basis of these findings, the case was diagnosed as Xp11 translocation RCC. This tumor massively recurred and led to the patient's death 2 years after the initial diagnosis. The utility of immunohistochemistry using antibodies against TFE3 in RCC occurring in young adults may be necessary for accurate diagnosis. PMID- 19298351 TI - Multicentric hyaline-vascular Castleman's disease treated by laparoscopic resection. PMID- 19298352 TI - Profound bicalutamide withdrawal syndrome in a hormone-refractory T4N1 prostate cancer permitting both salvage radiotherapy and cessation of hormonal therapy. PMID- 19298353 TI - Antifungal activity of amphotericin B, caspofungin and posaconazole on Candida albicans biofilms in intermediate and mature development phases. AB - The aim of this study was to examine the antifungal activity of amphotericin B, caspofungin and posaconazole on Candida albicans biofilms in the intermediate and mature development phases. Candida albicans biofilms, previously grown for either 24, 48 or 72 h in 96-well microtitre plates, were treated for 48 h with amphotericin B, caspofungin or posaconazole in increasing concentrations according to the respective minimal inhibitory concentration (MIC) determined for planktonic cells (1-128 x MIC). The biofilms were quantified using the mean optical density (OD) determined by XTT assay. Antifungal activities were expressed as percentage of reduction in OD of drug-treated biofilms compared to untreated biofilms. To test the fungicidal activity of antifungal agents, the unfixed biofilms were scraped off and seeded to Sabouraud agar. Caspofungin and amphotericin B showed higher activity against C. albicans biofilm grown for 24 h and 72 h (>or=50% reduction of OD) than biofilms grown for 48 h, whereas posaconazole showed similar, but reduced activity against all phases of C. albicans biofilm (5 BU mL(-1)) inhibitors, bleeding episodes are treated with bypassing agents such as activated prothrombin complex concentrates (APCCs) and recombinant activated factor VII (rFVIIa, NovoSeven; Novo Nordisk A/S, Bagsvaerd, Denmark). It is possible to administer bypassing therapy regularly to prevent haemorrhages, with the goal of limiting arthropathy and serious life- and limb-threatening bleeding. The data evaluating the efficacy and safety of this approach in patients with inhibitors are limited, consisting of results from one prospective trial and retrospective case reports. This report describes our experience with the prophylactic use of the APCC Factor Eight Inhibitor Bypassing Activity, Anti-Inhibitor Coagulant Complex, Vapor Heated (FEIBA; Baxter AG, Vienna, Austria). Data from patients at one treatment centre were retrospectively evaluated. Case records of six patients with haemophilia A or B and high-titre inhibitors were identified. When APCC was administered regularly, most patients exhibited a reduction in the numbers of haemorrhages, an improvement in orthopaedic status, and an improvement in quality of life. Prophylaxis with APCC can reduce haemorrhages and halt further joint deterioration in patients with haemophilia and inhibitors. PMID- 19298381 TI - Stability, efficacy, and safety of continuously infused sucrose-formulated recombinant factor VIII (rFVIII-FS) during surgery in patients with severe haemophilia. AB - Bolus injection (BI) of sucrose-formulated recombinant factor VIII (rFVIII-FS) is an approved treatment for haemophilia patients undergoing major surgery. Continuous infusion (CI) during surgery has potential benefits by providing steady administration of replacement factor to the patient, avoiding high peaks and low troughs. We tested the stability of rFVIII-FS under CI conditions and conducted a single-centre, open-label, phase III study to evaluate the efficacy and safety of CI using rFVIII-FS in haemophilia A patients undergoing surgery. Patients received bolus rFVIII-FS to achieve >or=80% FVIII levels 30-60 min presurgery, followed by CI of rFVIII-FS at a rate calculated to maintain haemostatic factor levels until days 8-10 post surgery. The rate of infusion was adjusted according to daily calculations derived from the actual clearance. The stability of rFVIII-FS was found to be appropriate for CI for 7 days under the same conditions as clinical settings. Fourteen patients (mean age 37.8 years) receiving on-demand FVIII treatment without a history of inhibitors underwent 15 surgical procedures including joint replacements, synovectomies, multiple tooth extractions, and cholecystectomy. Bleeding was similar to that observed in non haemophilia patients undergoing similar operations in the same department. Haemostasis during surgery was considered by the attending surgeons as 'excellent' or 'good' in all cases; study investigators rated all 15 cases as 'excellent' overall. There were no adverse events, including inhibitor formation, related to rFVIII-FS. rFVIII-FS was found to be suitable for use in CI in haemophilia A patients undergoing major surgery. PMID- 19298382 TI - Single-dose (270 microg kg(-1)) recombinant activated factor VII for the treatment and prevention of bleeds in haemophilia A patients with inhibitors: experience from seven European haemophilia centres. AB - Several studies have suggested that recombinant factor VIIa (rFVIIa) is effective and safe at doses >90 microg kg(-1). In March 2007, the European Medicines Agency approved the use of single-dose rFVIIa 270 microg kg(-1) for the treatment of mild-to-moderate bleeds in haemophilia patients with inhibitors. The aim of this study was to describe the use of single-dose rFVIIa in a real-life setting. In November 2007, seven haemophilia specialists from five European countries convened to share and discuss their experiences with the single-dose rFVIIa regimen within haemophilia A. Case histories of eight patients were discussed in this retrospective study. Six adult and two paediatric patients (age range, 19 months-40 years) were treated with single-dose rFVIIa for a variety of target joint bleeding, other bleeds and bleeding prevention. Treatment was successful in all the eight cases, with most patients requiring one dose to achieve bleeding resolution. No thrombotic or other safety concerns were raised by single-dose rFVIIa treatment. All patients and physicians preferred single-dose rFVIIa treatment to multiple injections; key benefits of single-dose rFVIIa treatment reported by patients and physicians included improved quality of life, greater convenience and ease of administration, improved compliance, faster control of bleeding, less injection-related pain and faster pain relief. In the patients reported here, single-dose rFVIIa 270 microg kg(-1) appears to be an effective and safe haemostatic treatment that improves the quality of life and convenience of treatment for patients. Such treatment might be of particular benefit for patients with difficult venous access or needle phobia. PMID- 19298383 TI - Experience with a third generation recombinant factor VIII concentrate (Advate) for immune tolerance induction in patients with haemophilia A. AB - The development of an inhibitor represents one of the most challenging complications in patients with haemophilia A. Optimal management is immune tolerance induction (ITI), typically through the administration of high doses of factor VIII (FVIII) concentrate. Among 12 patients who underwent ITI using Advate, a third-generation recombinant FVIII product that is free of animal and human protein additives, tolerance was achieved in nine (75%), including seven of 10 patients (70%) with high-titre inhibitors. ITI is ongoing in two patients and not yet successful; immune tolerance failed in the third patient. The median time to success was 4.0 months for group as a whole and for patients with high-titre inhibitors. Treatment was well tolerated, and no adverse events were observed. Advate was found to be equivalent to other FVIII products with regard to both ITI success rates and the incidence of adverse effects when used in these immune tolerance regimens. PMID- 19298384 TI - Inhibitor development: patient-determined risk factors. AB - The reasons that inhibitory factor VIII antibodies develop in only a fraction of patients with haemophilia A remain unclear, but studies of genetically related subjects have indicated that the immunological outcome of replacement therapy is to a large extent determined by patient-related risk factors. Non-genetic factors will also influence the inhibitor risk, since events challenging the immune system will elicit and stimulate immune regulatory processes with the potential of modifying the immune response. Further insight into the immunological pathways and risk factors involved will be important in order to better predict and prevent this complication. This review will briefly summarize the data obtained to date in unrelated and related subjects in the Malmo International Brother Study (MIBS) regarding genetic factors and discuss how these factors might interact with non-genetically determined factors and events. PMID- 19298385 TI - Current approaches in haemophilic arthropathy of the hip. AB - The hip is considered to be one of the main load bearing joints of the body. In the haemophilic patient joint bleeds can be catastrophic, leading to long-term joint degeneration and accompanying arthritis. In this review we explore the mechanisms of joint destruction, with particular consideration of the anatomy of the hip and how it may influence disease progression. We also review current strategies for treatment including hip replacement in the haemophilic patient and describe our experiences as a unit. Finally we evaluate future prospects in the management of hip disease in haemophilia. PMID- 19298386 TI - Evidence for beta-lactoglobulin involvement in vitamin D transport in vivo--role of the gamma-turn (Leu-Pro-Met) of beta-lactoglobulin in vitamin D binding. AB - Beta-lactoglobulin (LG) is a major bovine milk protein, containing a central calyx and a second exosite beyond the calyx to bind vitamin D; however, the biological function of LG in transporting vitamin D remains elusive. Crystallographic findings from our previous study showed the exosite to be located at the pocket between the alpha-helix and beta-strand I. In the present study, using site-directed mutagenesis, we demonstrate that residues Leu143, Pro144 and Met145 in the gamma-turn loop play a crucial role in the binding. Further evidence is provided by the ability of vitamin D(3) to block the binding of a specific mAb in the gamma-turn loop. Using the mouse (n = 95) as an animal model, we initially demonstrated that LG is a major fraction of milk proteins responsible for uptake of vitamin D. Most interestingly, dosing mice with LG supplemented with vitamin D(3) revealed that native LG containing two binding sites gave a saturated concentration of plasma 25-hydroxyvitamin D at a dose ratio of 2 : 1 (vitamin D(3)/LG), whereas heated LG containing one exosite (lacking a central calyx) gave a ratio of 1 : 1. We have demonstrated for the first time that LG has a functional advantage in the transport of vitamin D, indicating that supplementing milk with vitamin D effectively enhances its uptake. PMID- 19298387 TI - Allosteric and binding properties of Asp1-Glu382 truncated recombinant human serum albumin--an optical and NMR spectroscopic investigation. AB - Human serum albumin (HSA) is known for its exceptional ligand-binding capacity; indeed, its modular domain organization provides a variety of ligand-binding sites. Its flexible modular structure involves more than the immediate vicinity of the binding site(s), affecting the ligand-binding properties of the whole protein. Here, biochemical characterization by (1)H-NMR relaxometry and optical spectroscopy of a truncated form of HSA (tHSA) encompassing domains I and II (Asp1-Glu382) is reported. Removal of the C-terminal domain III results in a number of contacts that involve domain I (containing the heme site) and domain II (containing the warfarin site) being lost; however, the allosteric linkage between heme and warfarin sites is maintained. tHSA shows a nuclear magnetic relaxation dispersion profile similar to that of HSA, and displays increased affinity for ibuprofen, warfarin, and heme, suggesting that the fold is preserved. Moreover, the allosteric properties that make HSA a peculiar monomeric protein and account for the regulation of ligand-binding modes by heterotropic interactions are maintained after removal of domain III. Therefore, tHSA is a valuable model with which to investigate allosteric properties of HSA, allowing independent analysis of the linkages between different drug-binding sites. PMID- 19298388 TI - Dynamics of thromboxane level changes during early phase of allograft reperfusion. AB - BACKGROUND: Thromboxane (Tx) is a metabolite of arachidonic acid, which exerts a significant influence on kidney homeostasis, and may be involved in the pathogenesis of allograft rejection. The aim of this study was to: examine the dynamics of TxB2 changes during early phase of kidney allograft reperfusion, and analyze whether the observed changes in the concentrations and direction of TxB2 changes, are associated with post-transplant graft function. METHODS: Sixty-nine transplant recipients were divided into early, slow and delayed graft function group. Blood samples were collected directly before and during first the five minutes of allograft reperfusion. TxB2 concentrations were measured using ELISA. Creatinine and GFR levels were measured on the first, fifth, and 10th post transplant day. RESULTS: The results demonstrated that during reperfusion significant differences in TxB2 concentrations occur in all groups. Moreover, significant differences in the concentrations, as well as in the dynamics of TxB2 changes between patients with immediate graft function, and individuals with allograft activation problems, were noticed. These differences were associated with post-transplant graft function. CONCLUSIONS: Human renal transplantations are accompanied by changes in TxB2 concentrations, and the dynamics of TxB2 changes is associated with early post-transplant graft function. Our results also highlight TxB2 as a potential pre-transplant marker of post-transplant allograft function. PMID- 19298389 TI - A direct association of polyomavirus BK viruria with deterioration of renal allograft function in renal transplant patients. AB - BACKGROUND: Polyomavirus BK virus (BKV) causes a BKV-associated nephropathy (BKVAN), frequently causing allograft dysfunction in renal transplant recipients. As BK viruria is a surrogate marker for early detection of BKVAN, the aim of this study was to clarify an association between BK viruria and allograft dysfunction in renal transplant recipients. METHODS: One hundred and six renal transplant recipients with average 5.9-yr transplant duration received screening for quantification of BK viruria detected by real time polymerase chain reaction and were followed up for 12 months. RESULTS: Twenty-six patients (25%) had detectable BK viruria. In comparison of the patients without BK viruria, more patients in the BK viruria group were treated with steroids and had a past history of acute rejection. There was no difference in sex, age, transplant duration, allograft type and previous cytomegalovirus infection. During follow-up, the patients with BK viruria had higher serum creatinine levels at the sixth, ninth and 12th month. Multiple logistic regression analysis revealed that BK viruria was the only risk factor for more than 25% or 50% rise of serum creatinine level above baseline at the end of one yr follow-up. CONCLUSIONS: BK viruria alone is associated with allograft dysfunction and early intervention is indicated. PMID- 19298390 TI - Aortoiliac surgery concomitant with kidney transplantation: a single center experience. AB - INTRODUCTION: Aortoiliac pathology in kidney allograft recipients is not rare but treatment timing is controversial. As most publications on this topic are case reports its difficult to evaluate long-term outcomes of those clinical challenges. Herein we report long-term results of these procedures. METHODS: From 1970 to 2006, 1544 kidney transplants were performed in our center. Thirty patients underwent aortoiliac surgery simultaneously with kidney transplantation. We analyzed their clinical records to come up with outcomes of these complex clinical challenges. RESULTS: Vascular pathology was distributed as following: 19 stenoses treated with endarterectomy (15), aortoiliac bypass (two), aorto-bi iliac bypass (one) and aorto-bifemoral bypass (one); and 11 aneurysms treated with arterioplasty (four), aorto-bi-iliac bypass (four) and iliac-iliac bypass (three). In 24 cases (80%) the necessity of vascular surgery was established intraoperatively as vessels conditions did not permit safe anastomoses and jeopardized graft survival. Mean follow-up was 59 months (12-125). Five (16.7%) grafts were lost and three (10%) patients died in the first postoperative month: acute myocardial infarction (two) and non-viable kidney (one). Three patients died six, seven and 10 yr after the procedure. Nineteen patients are currently well with functioning grafts. CONCLUSIONS: Surgical correction of aortoiliac pathology may be performed simultaneously with kidney transplantation with acceptable outcome. This complex surgery can be performed in centers with experienced vascular surgeons. Specific vascular imaging should be performed regularly on patient at risk of aortoiliac disease before insertion and while on waiting list. PMID- 19298391 TI - Transport of interleukin-1 across cerebromicrovascular endothelial cells. AB - BACKGROUND AND PURPOSE: The inflammatory cytokine interleukin-1 (IL-1) has profound actions in the brain, causing neuronal cell death and exacerbating brain damage. While circulating levels are normally low, IL-1 can be produced on the vascular side of the brain endothelium, and within the brain. The naturally occurring IL-1 receptor antagonist has been administered peripherally in a Phase II trial in acute stroke patients; understanding how IL-1 and IL-1 receptor antagonist penetrate the brain is, therefore, of considerable importance. EXPERIMENTAL APPROACH: An in vitro blood-brain barrier model was generated by co culture of porcine brain microvascular endothelial cells with astrocytes. The mechanisms of transcellular transport of IL-1beta and IL-1 receptor antagonist were characterized in this model, using endocytosis inhibitors and IL-1 receptor blocking antibodies. KEY RESULTS: Transcellular IL-1beta and IL-1 receptor antagonist transport was temperature-dependent and IL-1beta was transported with higher affinity than IL-1 receptor antagonist. IL-1beta inhibited IL-1 receptor antagonist transport more potently than IL-1 receptor antagonist inhibited IL 1beta transport. Transport of IL-1beta and IL-1 receptor antagonist was not via adsorptive-mediated endocytosis, although inhibition of microtubule assembly significantly attenuated transport of both cytokines. An antibody directed to the type II IL-1 receptor significantly reduced IL-1beta transport. CONCLUSIONS AND IMPLICATIONS: These results are consistent with IL-1 and IL-1 receptor antagonist being transported across cultured cerebromicrovascular endothelial cells and suggest that IL-1beta transport may occur via a type II IL-1 receptor-dependent mechanism. Understanding IL-1 transport into the brain may have benefits, particularly in enhancing penetration of IL-1 receptor antagonist into the brain. PMID- 19298393 TI - The phosphodiesterase-5 inhibitor vardenafil improves cardiovascular dysfunction in experimental diabetes mellitus. AB - BACKGROUND AND PURPOSE: Patients with diabetes mellitus exhibit generalized endothelial and cardiac dysfunction with decreased nitric oxide production. Elevated intracellular cyclic guanosine monophosphate (cGMP) levels contribute to an effective cardioprotection in different pathophysiological conditions. In this study, we investigated whether chronic treatment with the phosphodiesterase-5 inhibitor vardenafil could improve diabetic cardiovascular dysfunction by up regulating the nitric oxide-cGMP pathway in the vessel wall and myocardium. EXPERIMENTAL APPROACH: Diabetes was induced in young rats by a single intraperitoneal injection of streptozotocin (60 mg x kg(-1)). In the treatment group, vardenafil (10 mg x kg(-1) x day(-1)) was given orally for 8 weeks. Diabetic control animals received vehicle for the same time. Left ventricular pressure-volume relations were measured by using a microtip Millar pressure volume conductance catheter, and indexes of contractility, such as the slope of end-systolic pressure-volume relationship (E(max)) and preload recruitable stroke work (PRSW), were calculated. In organ bath experiments for isometric tension with rings of isolated aortae, endothelium-dependent and independent vasorelaxation was investigated by using acetylcholine and sodium nitroprusside. KEY RESULTS: When compared with the non-diabetic controls, diabetic rats showed increased myocardial and vascular transforming growth factor-beta1 expression, impaired left ventricular contractility (impairment of E(max) by 53%, PRSW by 40%; P < 0.05) and vascular dysfunction. Treatment with vardenafil resulted in higher cGMP levels, reduced transforming growth factor-beta1 expression, significantly improved cardiac function (improvement of E(max) by 95%, PRSW by 69%; P < 0.05) and greater vasorelaxation to acetylcholine and sodium nitroprusside in aortae from diabetic animals. CONCLUSIONS AND IMPLICATIONS: Our results demonstrate that impaired vascular cGMP signalling contributes to the development of diabetic vascular and cardiac dysfunction, which can be prevented by chronic phosphodiesterase-5 inhibition. PMID- 19298394 TI - Molecular basis for amino acid sensing by family C G-protein-coupled receptors. AB - Family C of human G-protein-coupled receptors (GPCRs) is constituted by eight metabotropic glutamate receptors, two gamma-aminobutyric acid type B (GABA(B1-2)) subunits forming the heterodimeric GABA(B) receptor, the calcium-sensing receptor, three taste1 receptors (T1R1-3), a promiscuous L-alpha;-amino acid receptor G-protein-coupled receptor family C, group 6, subtype A (GPRC6A) and seven orphan receptors. Aside from the orphan receptors, the family C GPCRs are dimeric receptors characterized by a large extracellular Venus flytrap domain which bind the endogenous agonists. Except from the GABA(B1-2) and T1R2-3 receptor, all receptors are either activated or positively modulated by amino acids. In this review, we outline mutational, biophysical and structural studies which have elucidated the interaction of the amino acids with the Venus flytrap domains, molecular mechanisms of receptor selectivity and the initial steps in receptor activation. PMID- 19298395 TI - Myrtucommulone, a natural acylphloroglucinol, inhibits microsomal prostaglandin E(2) synthase-1. AB - BACKGROUND AND PURPOSE: The selective inhibition of prostaglandin (PG)E(2) formation via interference with microsomal PGE(2) synthase (mPGES)-1 could have advantages in the treatment of PGE(2)-associated diseases, such as inflammation, fever and pain, compared with a general suppression of all PG biosynthesis, provided by inhibition of cyclooxygenase (COX)-1 and 2. Here, we addressed whether the naturally occurring acylphloroglucinol myrtucommulone (MC) from Myrtus communis L. (myrtle) affected mPGES-1. EXPERIMENTAL APPROACH: The effect of MC on PGE(2) formation was investigated in a cell-free assay by using microsomal preparations of interleukin-1beta-stimulated A549 cells as the source of mPGES-1, in intact A549 cells, and in lipopolysaccharide-stimulated human whole blood. Inhibition of COX-1 and COX-2 activity in cellular and cell-free assays was assessed by measuring 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid and 6-oxo PGF(1alpha) formation. KEY RESULTS: MC concentration-dependently inhibited cell-free mPGES-1-mediated conversion of PGH(2) to PGE(2) (IC(50) = 1 micromol x L(-1)). PGE(2) formation was also diminished in intact A549 cells as well as in human whole blood at low micromolar concentrations. Neither COX-2 activity in A549 cells nor isolated human recombinant COX-2 was significantly affected by MC up to 30 micromol x L(-1), and only moderate inhibition of cellular or cell-free COX-1 was evident (IC(50) > 15 micromol x L(-1)). CONCLUSIONS AND IMPLICATIONS: MC is the first natural product to inhibit mPGES-1 that efficiently suppresses PGE(2) formation without significant inhibition of the COX enzymes. This provides an interesting pharmacological profile suitable for interventions in inflammatory disorders, without the typical side effects of coxibs and non-steroidal anti-inflammatory drugs. PMID- 19298396 TI - Molecular identification of spotted fever group Rickettsiae in ticks collected in central China. PMID- 19298397 TI - Effects of Rickettsia conorii infection on the survival of Rhipicephalus sanguineus ticks. PMID- 19298398 TI - Comparative study of pediculicidal effect of medical plants. PMID- 19298399 TI - Diagnostic value of ocular examination in Mediterranean spotted fever. PMID- 19298400 TI - Rickettsia rickettsii, Rickettsia felis and Rickettsia sp. TwKM03 infecting Rhipicephalus sanguineus and Ctenocephalides felis collected from dogs in a Brazilian spotted fever focus in the State of Rio De Janeiro/Brazil. PMID- 19298401 TI - Comparative ompA gene sequence analysis of Rickettsia felis-like bacteria detected in Haemaphysalis sulcata ticks and isolated in the mosquito C6/36 cell line. PMID- 19298402 TI - Isolated fever in horses: a new case of equine anaplasmosis in France. PMID- 19298403 TI - Seroprevalence of Coxiella burnetii antibodies in human immunodeficiency virus positive patients in Jacarepagua, Rio de Janeiro, Brazil. PMID- 19298404 TI - First detection of Anaplasma phagocytophilum-like DNA in the French izard Rupricapra pyrenaica. PMID- 19298405 TI - Dynamics of natural infection by Anaplasma phagocytophilum in a dairy cattle herd in Brittany, France. PMID- 19298406 TI - Prospective comparison of five mediators of the systemic response after high intensity focused ultrasound and targeted cryoablation for localized prostate cancer. AB - OBJECTIVE: To evaluate whether high-intensity focused ultrasound (HIFU) is less invasive than targeted cryoablation of the prostate (TCAP), as experimental studies suggest that the acute-phase reaction is proportional to surgery-induced tissue damage. PATIENTS AND METHODS: Between May 2004 and December 2007, 127 consecutive patients undergoing HIFU (71) or TCAP (56) in our departments were assessed prospectively. Blood samples were collected 24 h before (T0), during surgery (T1), at the end of anaesthesia (T2), and at 12 (T3), 24 (T4) and 36 h after surgery (T5). The extent of the systemic response to surgery-induced tissue trauma was measured by assessing the levels of acute-phase markers tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10, C-reactive protein (CRP) and serum amyloid A (SAA), at all sampling times in all patients. RESULTS: The median (interquartile range) age in the HIFU and TCAP groups was 70 (62-77) and 68 (59-75) years, respectively (P = 0.4). Baseline levels (T0) of TNF alpha, IL-6, IL-10, CRP and SAA were comparable in both groups. The levels of all four markers increased during both procedures; the median values were higher for TCAP at T2 (P = 0.02, 0.001, <0.001 and 0.06), T3 (P < 0.001, 0.009, 0.003 and 0.001), T4 (P = 0.007, <0.001, 0.005 and <0.001) and T5 (P < 0.001, 0.004, <0.001 and 0.02), respectively. IL-6 was also higher for TCAP at T1 (P = 0.03). IL-10 did not change at the different sampling times. CONCLUSION: The tissue trauma and associated invasiveness of HIFU is less than that of TCAP, based on the variables objectively measured in this study. PMID- 19298407 TI - Laparo-endoscopic single-site surgery in urology: is robotics the missing link? PMID- 19298408 TI - Open radical retropubic prostatectomy gives favourable surgical and functional outcomes after transurethral resection of the prostate. AB - OBJECTIVES: To assess the peri- and postoperative outcome of patients treated with open radical retropubic prostatectomy (RRP) for prostate cancer and who had previously undergone transurethral resection of the prostate (TURP). PATIENTS AND METHODS: Prospectively collected data from a consecutive series of 1760 patients who had RRP between July 2003 and June 2007 at our institution were used to retrospectively match 62 cases (with previous TURP) with the same number of controls (without previous TURP). Matching variables were patient age, body mass index, prostate volume, preoperative total prostate-specific antigen (PSA) level, Gleason score, pathological stage, and intraoperative nerve-sparing procedure. Complete 1-year follow-up data were available for all patients. All collected data on surgery and perioperative complications were analysed. Functional outcome data at the 1-year follow-up were evaluated by applying an institutional questionnaire. Sexual function was assessed using the abbreviated International Index of Erectile Function-5 questionnaire, and urinary control was evaluated by defining complete urinary control as no pad usage. RESULTS: The rate of complete urinary control rate in cases and controls was similar (81% vs 82%). When nerves were spared, 60% (15/25) of patients in either group were capable of sexual intercourse. The overall positive surgical margin rate was insignificantly higher in cases (19% vs 13, P>0.05). After 1 year of follow-up the biochemical recurrence rate (PSA>0.04 ng/mL) did not differ significantly in patients who had RRP after TURP vs RRP alone (six of 62, 10%, vs five of 62, 8%; P=0.77). CONCLUSIONS: RRP for prostate cancer in patients who have had previous TURP does not result in a higher perioperative complication rate, or a worse functional outcome. PMID- 19298409 TI - Developments in technique and technology: the effect on the results of percutaneous nephrolithotomy for staghorn calculi. AB - OBJECTIVE: To review the development of the technique of percutaneous nephrolithotomy (PCNL), for ease of learning and development of instrumentation for staghorn calculi at our centre since 1991, and to assess the results and outcomes. PATIENTS AND METHODS: We retrospectively analysed the hospital records of 773 patients (632 males and 141 females, 834 renal units) who underwent PCNL for staghorn calculi at our centre from January 1991 to August 2008. We divided the patients into three groups depending on the changes in treatment policy, global trends and advances in equipment as follows: the first 200 cases (group I) from January 1991 to December 1996 (216 renal units); the next 200 (group II) from January 1997 to December 2001 (212 renal units); and the last 373 (group III) from January 2002 to August 2008 (406 renal units). RESULTS: The mean (sd, range) operative duration in groups I, II and III, respectively, were 138.2 (52.7, 60-310), 121.4 (42.8, 70-250) and 112.5 (51.5, 55-310) min; the decrease in haemoglobin level was 3.2, 2.6 and 1.6 g/dL, respectively, and continued to decrease with improvements in technique. With increasing experience, the number of stages required for stone clearance and the number of tracts required decreased exponentially. Most of the severe complications occurred early in our experience. The stone clearance rate in groups I, II and III was 81%, 86% and 93%, respectively, after completing the procedure; the overall clearance rate with observation/auxiliary procedures was 86%, 89% and 96%, respectively. The mean hospital stay for groups I, II and III was 11.1 (3.9, 7-25), 9.5 (3.4, 5-22) and 7.1 (3.6, 4-28) days, respectively. CONCLUSION: The percutaneous management of staghorn calculi requires considerable expertise. Our data suggest that 'multiperc' PCNL is difficult to learn and requires experience. Although over the years our results improved, complete clearance remains a challenge. A constant review and application of newer techniques and results will improve the overall clearance rates further. PMID- 19298410 TI - Increased vascular endothelial growth factor expression in patients with bladder pain syndrome/interstitial cystitis: its association with pain severity and glomerulations. AB - OBJECTIVE: To determine the angiogenic profiles in the bladder of patients with bladder pain syndrome (BPS)/interstitial cystitis (IC), and to evaluate the relationship between these profiles and associated clinical features including pelvic pain and glomerulations. PATIENTS AND METHODS: Angiogenesis and angiogenic components are important in chronic inflammatory disease. High levels of vascular endothelial growth factor (VEGF) have been shown to induce immature angiogenesis, where microvessels have insufficient coverage of pericytes, resulting in haemorrhagic vessels. Biopsy specimens from 30 patients with BPS/IC and glomerulations, and 10 control patients, were examined immunohistochemically for VEGF expression, microvessel density (MVD) and immature microvessels. Pericyte coverage of microvessels in the specimens was used as an indicator of mature microvessels, and pericytes were identified by double-immunohistochemistry for CD34 and alpha-smooth muscle actin. The microvessel pericyte coverage index (MPI) was calculated as the ratio of mature vessels to total vessels. We also assessed the relationship between these angiogenic profiles and associated clinical features including pain and glomerulations. RESULTS: VEGF expression in the lamina propria was significantly higher in BPS/IC than in control samples (50% vs 10%, P < 0.05). Among patients with BPS/IC, VEGF expression was significantly higher in those with severe pain than in those with mild pain (78% vs 38%, P < 0.05). The MPI was significantly lower in BPS/IC than in control samples (23% vs 35%, P < 0.05), whereas MVD did not differ significantly between BPS/IC and control samples. CONCLUSIONS: There is increased VEGF and immature vascularization in patients with BPS/IC, and VEGF expression is associated with the degree of pain described by patients. Taken together, VEGF might contribute to pain and promote the formation of immature vessels in BPS/IC, and the increased immature vascularization might have a role in glomerulations in patients with BPS/IC. PMID- 19298411 TI - Exposure to Agent Orange is a significant predictor of prostate-specific antigen (PSA)-based recurrence and a rapid PSA doubling time after radical prostatectomy. AB - OBJECTIVE: To investigate and report the clinicopathological characteristics and outcomes after radical prostatectomy (RP) in patients with prostate cancer and previous exposure to Agent Orange (AO), particularly in relationship to race. PATIENTS AND METHODS: In 1495 veterans who had undergone RP the clinicopathological characteristics, biochemical progression rates, and prostate specific antigen (PSA) doubling time (DT) after recurrence between AO-exposed and unexposed men were compared using logistic and linear regression and Cox proportional hazards analyses, and stratified by race. RESULTS: The 206 (14%) men with AO exposure were more likely to be black (P = 0.001), younger (P < 0.001), treated more recently (P < 0.001), have a higher body mass index (P = 0.001), have clinical stage T1 disease (P < 0.001), and have lower preoperative PSA levels (P = 0.001). After adjusting for several clinical characteristics, AO exposure was not significantly related to adverse pathological features but was significantly associated with biochemical progression risk (relative risk 1.55, 95% confidence interval 1.15-2.09, P = 0.004) and shorter PSADT (P < 0.001) after recurrence (8.2 vs 18.6 months). When stratified by race, these associations were present and similar in both races, with no significant interaction between race and AO exposure for predicting biochemical recurrence or mean adjusted PSADT (P interaction >0.20). CONCLUSIONS: Patients with AO exposure and treated with RP were more likely to be black, present with lower risk features, have an increased risk of biochemical progression, and shorter PSADT after recurrence. When stratified by race, the association between AO exposure and poor outcomes was present in both races. These findings suggest that among selected men who choose RP, AO exposure might be associated with more aggressive prostate cancer. PMID- 19298412 TI - Short-term administration of prulifloxacin in patients with nonmuscle-invasive bladder cancer: an effective option for the prevention of bacillus Calmette Guerin-induced toxicity? AB - OBJECTIVE: To determine whether the new fluoroquinolone prulifloxacin might improve tolerance to Bacillus Calmette-Guerin (BCG) intravesical therapy in patients with bladder cancer. PATIENTS AND METHODS: A series of 72 patients with intermediate- or high-risk nonmuscle-invasive bladder cancer were enrolled in this prospective, randomized, open-label, controlled clinical trial performed at a single tertiary care institution. After complete transurethral resection, patients were randomized to receive induction treatment with BCG and three capsules of prulifloxacin 600 mg or no prophylactic treatment (control group). Adverse events (AEs) were self-recorded by the patients after each instillation and classified by the investigator according to a classification grid considering account duration and intensity. Cystoscopy findings at 3 and 6 months were also recorded. RESULTS: There was no significant difference in baseline symptoms between the groups. Overall, there was a significant decrease in the percentage of patients with at least one AE between instillations in prulifloxacin-treated group. The proportion of patients with moderate to severe AEs after the fourth instillation was significantly less in the prulifloxacin-treated group. There was a significant effect of prulifloxacin on the need for anti-tuberculosis treatment. More patients in the control group stopped or delayed the full induction course of BCG instillations (34% vs 19%, P=0.04). Recurrence rates were not affected by prulifloxacin treatment. CONCLUSION: Prulifloxacin reduces the incidence of moderate to severe AEs from BCG intravesical therapy in patients with nonmuscle-invasive bladder cancer, improving compliance to the induction BCG course. These preliminary findings warrant further clinical research. PMID- 19298414 TI - Obesity metaphors: how beliefs about the causes of obesity affect support for public policy. AB - CONTEXT: Relatively little is known about the factors shaping public attitudes toward obesity as a policy concern. This study examines whether individuals' beliefs about the causes of obesity affect their support for policies aimed at stemming obesity rates. This article identifies a unique role of metaphor-based beliefs, as distinct from conventional political attitudes, in explaining support for obesity policies. METHODS: This article used the Yale Rudd Center Public Opinion on Obesity Survey, a nationally representative web sample surveyed from the Knowledge Networks panel in 2006/07 (N = 1,009). The study examines how respondents' demographic and health characteristics, political attitudes, and agreement with seven obesity metaphors affect support for sixteen policies to reduce obesity rates. FINDINGS: Including obesity metaphors in regression models helps explain public support for policies to curb obesity beyond levels attributable solely to demographic, health, and political characteristics. The metaphors that people use to understand rising obesity rates are strong predictors of support for public policy, and their influence varies across different types of policy interventions. CONCLUSIONS: Over the last five years, the United States has begun to grapple with the implications of dramatically escalating rates of obesity. Individuals use metaphors to better understand increasing rates of obesity, and obesity metaphors are independent and powerful predictors of support for public policies to curb obesity. Metaphorical reasoning also offers a potential framework for using strategic issue framing to shift support for obesity policies. PMID- 19298413 TI - Personality traits as prospective predictors of suicide attempts. AB - OBJECTIVE: To examine higher order personality factors of negative affectivity (NA) and disinhibition (DIS), as well as lower order facets of impulsivity, as prospective predictors of suicide attempts in a predominantly personality disordered sample. METHOD: Data were analyzed from 701 participants of the Collaborative Longitudinal Personality Disorders Study with available follow-up data for up to 7 years. Cox proportional hazards regression analyses was used to examine NA and DIS, and facets of impulsivity (e.g. urgency, lack of perseverance, lack of premeditation and sensation seeking), as prospective predictors of suicide attempts. RESULTS: NA, DIS and all facets of impulsivity except for sensation seeking were significant in univariate analyses. In multivariate models which included sex, childhood sexual abuse, course of major depressive disorder and substance use disorders, only NA and lack of premeditation remained significant in predicting suicide attempts. DIS and the remaining impulsivity facets were not significant. CONCLUSION: NA emerged as a stronger and more robust predictor of suicide attempts than DIS and impulsivity, and warrants greater attention in suicide risk assessment. Distinguishing between facets of impulsivity is important for clinical risk assessment. PMID- 19298415 TI - Reducing obesity: motivating action while not blaming the victim. AB - CONTEXT: The rise in obesity in the United States may slow or even reverse the long-term trend of increasing life expectancy. Like many risk factors for disease, obesity results from behavior and shows a social gradient. Especially among women, obesity is more common among lower-income individuals, those with less education, and some ethnic/racial minorities. METHODS: This article examines the underlying assumptions and implications for policy and the interventions of the two predominant models used to explain the causes of obesity and also suggests a synthesis that avoids "blaming the victim" while acknowledging the role of individuals' health behaviors in weight maintenance. FINDINGS: (1) The medical model focuses primarily on treatment, addressing individuals' personal behaviors as the cause of their obesity. An underlying assumption is that as independent agents, individuals make informed choices. Interventions are providing information and motivating individuals to modify their behaviors. (2) The public health model concentrates more on prevention and sees the roots of obesity in an obesogenic environment awash in influences that lead individuals to engage in health-damaging behaviors. Interventions are modifying environmental forces through social policies. (3) There is a tension between empowering individuals to manage their weight through diet and exercise and blaming them for failure to do so. Patterns of obesity by race/ethnicity and socioeconomic status highlight this tension. (4) Environments differ in their health-promoting resources; for example, poorer communities have fewer supermarkets, more fast food outlets, and fewer accessible and safe recreational opportunities. CONCLUSIONS: A social justice perspective facilitates a synthesis of both models. This article proposes the concept of "behavioral justice" to convey the principle that individuals are responsible for engaging in health-promoting behaviors but should be held accountable only when they have adequate resources to do so. This perspective maintains both individuals' control and accountability for behaviors and society's responsibility to provide health-promoting environments. PMID- 19298416 TI - Schools and obesity prevention: creating school environments and policies to promote healthy eating and physical activity. AB - CONTEXT: Research consistently shows that the majority of American children do not consume diets that meet the recommendations of the Dietary Guidelines for Americans, nor do they achieve adequate levels of daily physical activity. As a result, more children are overweight today than at any other time in U.S. history. Schools offer many opportunities to develop strategies to prevent obesity by creating environments in which children eat healthfully and engage regularly in physical activity. METHODS: This article discusses the role of schools in obesity prevention efforts. Current issues in schools' food and physical activity environments are examined, as well as federal, state, and local policies related to food and physical activity standards in schools. The article is organized around four key areas: (1) school food environments and policies, (2) school physical activity environments and policies, (3) school body mass index measurements, and (4) school wellness policies. Recommendations for accelerating change also are addressed. FINDINGS: The article found that (1) competitive foods (foods sold outside of federally reimbursed school meals) are widely available in schools, especially secondary schools. Studies have related the availability of snacks and drinks sold in schools to students' high intake of total calories, soft drinks, total fat and saturated fat, and lower intake of fruits and vegetables; (2) physical activity can be added to the school curriculum without academic consequences and also can offer physical, emotional, and social benefits. Policy leadership has come predominantly from the districts, then the states, and, to a much lesser extent, the federal government; (3) few studies have examined the effectiveness or impact of school-based BMI measurement programs; and (4) early comparative analyses of local school wellness policies suggest that the strongest policies are found in larger school districts and districts with a greater number of students eligible for a free or reduced-price lunch. CONCLUSIONS: Studies show that schools have been making some progress in improving the school food and physical activity environments but that much more work is needed. Stronger policies are needed to provide healthier meals to students at schools; limit their access to low-nutrient, energy-dense foods during the school day; and increase the frequency, intensity, and duration of physical activity at school. PMID- 19298417 TI - Addressing obesity in the workplace: the role of employers. AB - CONTEXT: Employers have pursued many strategies over the years to control health care costs and improve care. Disappointed by efforts to manage costs through the use of insurance-related techniques (e.g., prior authorization, restricted provider networks), employers have also begun to try to manage health by addressing their employees' key lifestyle risks. Reducing obesity (along with tobacco use and inactivity) is a priority for employers seeking to lower the incidence and severity of chronic illness and the associated demand for health services. METHODS: This article describes the employer's perspective on the cost impact of obesity, discusses current practices in employer-sponsored wellness and weight management programs, provides examples from U.S. companies illustrating key points of employers' leverage and opportunities, and suggests policy directions to support the expansion of employers' initiatives, especially for smaller employers. FINDINGS: Researchers and policymakers often overlook the extensive efforts and considerable impact of employer-sponsored wellness and health improvement programs. Greater focus on opportunities in the workplace is merited, however, for the evidence base supporting the economic and health impacts of employer-sponsored health promotion and wellness is growing, although not as quickly as the experience base of large employers. CONCLUSIONS: Public and private employers can serve their own economic interests by addressing obesity. Health care organizations, particularly hospitals, as well as public employers can be important role models. Policy development is needed to accelerate change, especially for smaller employers (those with fewer than 500 employees), which represent the majority of U.S. employers and are far less likely to offer health promotion programs. PMID- 19298418 TI - Physical activity and food environments: solutions to the obesity epidemic. AB - CONTEXT: Environmental, policy, and societal changes are important contributors to the rapid rise in obesity over the past few decades, and there has been substantial progress toward identifying environmental and policy factors related to eating and physical activity that can point toward solutions. This article is a status report on research on physical activity and food environments, and it suggests how these findings can be used to improve diet and physical activity and to control or reduce obesity. METHODS: This article summarizes and synthesizes recent reviews and provides examples of representative studies. It also describes ongoing innovative interventions and policy change efforts that were identified through conference presentations, media coverage, and websites. FINDINGS: Numerous cross-sectional studies have consistently demonstrated that some attributes of built and food environments are associated with physical activity, healthful eating, and obesity. Residents of walkable neighborhoods who have good access to recreation facilities are more likely to be physically active and less likely to be overweight or obese. Residents of communities with ready access to healthy foods also tend to have more healthful diets. Disparities in environments and policies that disadvantage low-income communities and racial minorities have been documented as well. Evidence from multilevel studies, prospective research, and quasi-experimental evaluations of environmental changes are just beginning to emerge. CONCLUSIONS: Environment, policy, and multilevel strategies for improving diet, physical activity, and obesity control are recommended based on a rapidly growing body of research and the collective wisdom of leading expert organizations. A public health imperative to identify and implement solutions to the obesity epidemic warrants the use of the most promising strategies while continuing to build the evidence base. PMID- 19298419 TI - A cross-sectional prevalence study of ethnically targeted and general audience outdoor obesity-related advertising. AB - CONTEXT: Commercial marketing is a critical but understudied element of the sociocultural environment influencing Americans' food and beverage preferences and purchases. This marketing also likely influences the utilization of goods and services related to physical activity and sedentary behavior. A growing literature documents the targeting of racial/ethnic and income groups in commercial advertisements in magazines, on billboards, and on television that may contribute to sociodemographic disparities in obesity and chronic disease risk and protective behaviors. This article examines whether African Americans, Latinos, and people living in low-income neighborhoods are disproportionately exposed to advertisements for high-calorie, low nutrient-dense foods and beverages and for sedentary entertainment and transportation and are relatively underexposed to advertising for nutritious foods and beverages and goods and services promoting physical activities. METHODS: Outdoor advertising density and content were compared in zip code areas selected to offer contrasts by area income and ethnicity in four cities: Los Angeles, Austin, New York City, and Philadelphia. FINDINGS: Large variations were observed in the amount, type, and value of advertising in the selected zip code areas. Living in an upper-income neighborhood, regardless of its residents' predominant ethnicity, is generally protective against exposure to most types of obesity-promoting outdoor advertising (food, fast food, sugary beverages, sedentary entertainment, and transportation). The density of advertising varied by zip code area race/ethnicity, with African American zip code areas having the highest advertising densities, Latino zip code areas having slightly lower densities, and white zip code areas having the lowest densities. CONCLUSIONS: The potential health and economic implications of differential exposure to obesity-related advertising are substantial. Although substantive legal questions remain about the government's ability to regulate advertising, the success of limiting tobacco advertising offers lessons for reducing the marketing contribution to the obesigenicity of urban environments. PMID- 19298420 TI - Innovative legal approaches to address obesity. AB - CONTEXT: The law is a powerful public health tool with considerable potential to address the obesity issue. Scientific advances, gaps in the current regulatory environment, and new ways of conceptualizing rights and responsibilities offer a foundation for legal innovation. METHODS: This article connects developments in public health and nutrition with legal advances to define promising avenues for preventing obesity through the application of the law. FINDINGS: Two sets of approaches are defined: (1) direct application of the law to factors known to contribute to obesity and (2) original and innovative legal solutions that address the weak regulatory stance of government and the ineffectiveness of existing policies used to control obesity. Specific legal strategies are discussed for limiting children's food marketing, confronting the potential addictive properties of food, compelling industry speech, increasing government speech, regulating conduct, using tort litigation, applying nuisance law as a litigation strategy, and considering performance-based regulation as an alternative to typical regulatory actions. Finally, preemption is an overriding issue and can play both a facilitative and a hindering role in obesity policy. CONCLUSIONS: Legal solutions are immediately available to the government to address obesity and should be considered at the federal, state, and local levels. New and innovative legal solutions represent opportunities to take the law in creative directions and to link legal, nutrition, and public health communities in constructive ways. PMID- 19298421 TI - Public health law and the prevention and control of obesity. AB - CONTEXT: Obesity constitutes a major public health challenge in the United States. Obesogenic environments have increased owing to the consumption of calorie-dense foods of low nutritional value and the reduction of daily physical activity (e.g., increased portion sizes of meals eaten in and out of the home and fewer physical activity requirements in schools). Policymakers and public health practitioners need to know the best practices and have the competencies to use laws and legal authorities to reverse the obesity epidemic. For instance, statutes and regulations at the federal, state, and local levels of government have been implemented to improve nutritional choices and access to healthy foods, encourage physical activity, and educate consumers about adopting healthy lifestyles. METHODS: In an effort to understand the application of laws and legal authorities for obesity prevention and control, in June 2008 the Centers for Disease Control and Prevention convened the National Summit on Legal Preparedness for Obesity Prevention and Control. An outcome of this summit will be the publication of the proceeding's white papers written by eight law and subject matter experts with substantive contributions from summit participants, which will identify actionable options that sectors and organizations at various jurisdictional levels can consider adopting. FINDINGS: Law has played a critical role in the control of chronic diseases and the behaviors that lead to them. The use of a systematic legal framework--the use of legislation, regulation, and policy to address the multiple factors that contribute to obesogenic environments -can assist in the development, implementation, and evaluation of a variety of legal approaches for obesity prevention and control. CONCLUSIONS: Although public health-focused legal interventions are in an early stage and the direct and indirect impact they may have on the obesity epidemic is not yet understood, efforts such as the summit and white papers should help determine potentially viable legal interventions and assess their impact on population-level change. PMID- 19298422 TI - Food prices and obesity: evidence and policy implications for taxes and subsidies. AB - CONTEXT: Pricing policies have been posited as potential policy instruments to address the increasing prevalence of obesity. This article examines whether altering the cost of unhealthy, energy-dense foods, compared with healthy, less dense foods through the use of fiscal pricing (tax or subsidy) policy instruments would, in fact, change food consumption patterns and overall diet enough to significantly reduce individuals' weight outcomes. METHODS: This article examined empirical evidence regarding the food and restaurant price sensitivity of weight outcomes based on a literature search to identify peer-reviewed English-language articles published between 1990 and 2008. Studies were identified from the Medline, PubMed, Econlit, and PAIS databases. The fifteen search combinations used the terms obesity, body mass index, and BMI each in combination with the terms price, prices, tax, taxation, and subsidy. FINDINGS: The studies reviewed showed that when statistically significant associations were found between food and restaurant prices (taxes) and weight outcomes, the effects were generally small in magnitude, although in some cases they were larger for low-socioeconomic status (SES) populations and for those at risk for overweight or obesity. CONCLUSIONS: The limited existing evidence suggests that small taxes or subsidies are not likely to produce significant changes in BMI or obesity prevalence but that nontrivial pricing interventions may have some measurable effects on Americans' weight outcomes, particularly for children and adolescents, low-SES populations, and those most at risk for overweight. Additional research is needed to be able to draw strong policy conclusions regarding the effectiveness of fiscal-pricing interventions aimed at reducing obesity. PMID- 19298423 TI - The perils of ignoring history: Big Tobacco played dirty and millions died. How similar is Big Food? AB - CONTEXT: In 1954 the tobacco industry paid to publish the "Frank Statement to Cigarette Smokers" in hundreds of U.S. newspapers. It stated that the public's health was the industry's concern above all others and promised a variety of good faith changes. What followed were decades of deceit and actions that cost millions of lives. In the hope that the food history will be written differently, this article both highlights important lessons that can be learned from the tobacco experience and recommends actions for the food industry. METHODS: A review and analysis of empirical and historical evidence pertaining to tobacco and food industry practices, messages, and strategies to influence public opinion, legislation and regulation, litigation, and the conduct of science. FINDINGS: The tobacco industry had a playbook, a script, that emphasized personal responsibility, paying scientists who delivered research that instilled doubt, criticizing the "junk" science that found harms associated with smoking, making self-regulatory pledges, lobbying with massive resources to stifle government action, introducing "safer" products, and simultaneously manipulating and denying both the addictive nature of their products and their marketing to children. The script of the food industry is both similar to and different from the tobacco industry script. CONCLUSIONS: Food is obviously different from tobacco, and the food industry differs from tobacco companies in important ways, but there also are significant similarities in the actions that these industries have taken in response to concern that their products cause harm. Because obesity is now a major global problem, the world cannot afford a repeat of the tobacco history, in which industry talks about the moral high ground but does not occupy it. PMID- 19298424 TI - The politics of obesity: a current assessment and look ahead. AB - CONTEXT: The continuing rise in obesity rates across the United States has proved impervious to clinical treatment or public health exhortation, necessitating policy responses. Nearly a decade's worth of political debates may be hardening into an obesity issue regime, comprising established sets of cognitive frames, stakeholders, and policy options. METHODS: This article is a survey of reports on recently published studies. FINDINGS: Much of the political discussion regarding obesity is centered on two "frames," personal-responsibility and environmental, yielding very different sets of policy responses. While policy efforts at the federal level have resulted in little action to date, state and/or local solutions such as calorie menu labeling and the expansion of regulations to reduce unhealthy foods at school may have more impact. CONCLUSIONS: Obesity politics is evolving toward a relatively stable state of equilibrium, which could make comprehensive reforms to limit rising obesity rates less feasible. Therefore, to achieve meaningful change, rapid-response research identifying a set of promising reforms, combined with concerted lobbying action, will be necessary. PMID- 19298427 TI - The clinical concept of epilepsy. AB - This article reviews the history of clinical concepts of epilepsy and its classification, especially in the last 100 years. Throughout its recorded history of 3 to 4 millennia, epilepsy has always been defined by its most dramatic symptoms, for example, falling, motor activity or loss of consciousness, but separation from other causes of the same paroxysmal symptoms has always proved challenging. For over a century there has been some semantic confusion whether to call the various paroxysms fits, convulsions, seizures, or epilepsies. Since the middle of the 19th century a great unresolved debate has continued about whether recurrent seizures or epilepsy should be viewed as a separable symptom of underlying brain disease or as one or more idiopathic diseases or syndromes, with an inherent age-related natural history; or indeed viewed as both a symptom and a disease. A major advance in the 20th century is that vascular theories of epilepsy, which reached their peak with Turner in 1907, have been replaced by electromagnetic discharges, based especially on the work of Todd, Jackson, Berger, Lennox, and the Gibbs, culminating eventually in new ILAE classifications of seizures (1981) and epilepsy syndromes (1989). However 21st century uncertainties about symptomatic versus idiopathic or cross-sectional (seizures) versus longitudinal (epilepsy) approaches to the problem very much reflect similar divergences of view a century ago. More attention is now being directed at interseizure events and processes which may lead either to remission or intractability with associated cognitive and psychosocial consequences. The search for the elusive essence, diathesis or predisposition to epilepsy, including seizure threshold, continues. PMID- 19298428 TI - Clinical neuropathology of the epilepsies in the 100 years of the ILAE (1909 2009). AB - The paper describes the history of the clinical neuropathology of the epilepsies in the last hundred years from microscopy to molecular neuropathology. The main focus is on the concepts of hippocampal sclerosis and its discussion of causative lesion or consequence of seizures, and the concept of developmental disturbances in respect to general epileptogenicity. Clinical neuropathology remains an important discipline in the future of epileptology and brain research especially in the area of molecular genetics. Neuropathology will help to understand the stages of epileptogenesis and the factors responsible for the progressive nature of the disease. PMID- 19298429 TI - Neurochemistry and epileptology. AB - This paper gives an account of the global evolution of (neuro-)chemistry in epileptology with an emphasis on the role of the International League Against Epilepsy (ILAE), which declared in its constitution a mission "to make the epilepsy-problem the object of special study and to make practical use of the results of such study." As Epilepsia is the scientific journal of the ILAE, the review emphasizes papers published in the journal. PMID- 19298430 TI - Clinical neurophysiology with special reference to the electroencephalogram. AB - In the nineteenth and early twentieth centuries, many important discoveries in nervous system structure and function involved the electrical properties of nerve tissue. The application of these advances, as well as those in electronic amplification and recording, led to the discovery of the human electroencephalogram (EEG) by Hans Berger, a German psychiatrist. Originally received with skepticism, the EEG became a subject of intense interest, and after World War II, became a leading clinical and experimental tool in neurology. Today, it remains important especially in the study and treatment of epilepsy. Though the EEG has also given rise to more sophisticated applications, these remain based on Berger's initial work, one of the great discoveries of medical history. PMID- 19298431 TI - A history of neuroimaging in epilepsy 1909-2009. AB - Profound advances in the field of clinical imaging in epilepsy occurred between 1909 and 2009, the century of the International League Against Epilepsy, and these are reviewed briefly in this paper. Initially imaging was carried out with plain x-ray, air encephalography, and angiography, and these techniques had a relatively minor role in epilepsy. Computerized tomographic (CT) scanning was introduced in 1971, and magnetic resonance imaging (MRI) a decade or so later, and both these technologies had an immediate and far-reaching impact on epilepsy. MRI techniques continued to evolve during the 1990s and profoundly influenced many aspects of epilepsy clinical practice. These structural imaging techniques revealed pathological lesions in large numbers of patients with hitherto cryptogenic epilepsy, widened the indications for surgical therapy, and improved our understanding of the pathogenesis and etiology of epilepsy. In recent years, the research focus has turned to fMRI but its impact on epilepsy currently is relatively small. Magnetic resonance spectroscopy (MRS), positron emission tomography (PET) and single photon emission computed tomography (SPECT) also have had a limited impact on clinical practice in epilepsy. PMID- 19298432 TI - Epilepsy, psychiatry, and neurology. AB - This article reviews the relationship between the psychiatry and neurology of epilepsy, especially in the last 100 years. Throughout most of its recorded history of 3 to 4 millennia epilepsy has been viewed as a supernatural or mental disorder. Although first suggested by Hippocrates in the 5th century B.C., the concept of epilepsy as a brain disorder only began to take root in the 17th and 18th centuries. The discipline of neurology emerged from "nervous disorders" or neuropsychiatry in the late 19th century, when vascular theories of epilepsy predominated. By the turn of the 19th century psychiatry and neurology were diverging and epilepsy remained to some extent in both disciplines. It was only in the middle of the 20th century with the development of electromagnetic theories of epilepsy that the concept of epilepsy per se as a neurological disorder was finally adopted in international classifications of disease. This was associated with a refined definition of the ictal, pre-, post-, and interictal psychological disorders of epilepsy, which have contributed to a renaissance of neuropsychiatry. At the beginning of the 21st century and the centenary of the ILAE psychiatry and neurology have been converging again, led in some respects by epilepsy, which has provided several useful models of mental illness and a bridge between the two disciplines. PMID- 19298433 TI - The history of status epilepticus and its treatment. AB - Despite the fact that status epilepticus was been recognized since antiquity, its existence was largely ignored until the mid-nineteenth century. In this review we cover the medical literature of status epilepticus from the late nineteenth century until the early 1970s when the modern era of status epilepticus began. We pay particular attention to the impact of the ILAE and its principal members on the understanding and awareness of status epilepticus. We also cover the evolution of treatment regimens advocated for status epilepticus from the late nineteenth century to the early 1970s when the benzodiazepines were established as first line treatments. PMID- 19298434 TI - Drug treatment of epilepsy in the century of the ILAE: the first 50 years, 1909 1958. AB - This paper provides a review of the drug treatment of epilepsy from 1909, the year of the foundation of the International League Against Epilepsy (ILAE), to 1958. It was during this period that modern approaches to the medicinal therapy of epilepsy were formulated and many new drugs were introduced. Of these, both phenytoin and phenobarbital continue to be widely used today. This was the period when scientific screening was also introduced into drug discovery. The leading figures in the ILAE played an important role in guiding the evolution of the treatment of epilepsy during this period. PMID- 19298435 TI - Drug treatment of epilepsy in the century of the ILAE: the second 50 years, 1959 2009. AB - The drug therapy of epilepsy evolved enormously in this 50 year period. Advances in therapeutics included the incorporation of pharmacokinetics into clinical practice, enormous advances in neurochemistry, a trend to antiepileptic drug monotherapy, better drug assessment, better understanding of therapeutic outcomes, and the recognition of the large epilepsy treatment gap in many countries. An unprecedented range of new drugs was introduced in this period. Before 1989, these included carbamazepine, valproate, ethosuximide, and the benzodiazepines. Since 1989, 13 more new drugs have been licensed and marketed and there are others in the pipeline. The International League Against Epilepsy and its leading figures have played an important role in these developments. In this period, too, there has been a rapid expansion in research and development within the pharmaceutical industry and a rise in the value of the antiepileptic drug market. In parallel, governmental regulation of pharmaceuticals has greatly increased. To what extent the overall prognosis of epilepsy has improved as a result of these activities is an interesting and perplexing question. PMID- 19298436 TI - Epilepsy surgery: historical highlights 1909-2009. AB - This review focuses on some historical highlights of the surgery of epilepsy, beginning with the reports of Horsley, Krause, and Cushing to which appeared in 1909, the year that The International League Against Epilepsy (ILAE) was inaugurated. We then outline key contributions from Europe and North America, and examine particularly the evolution of our understanding of temporal lobe seizures, which have now become the most common form of epilepsy amenable to surgical cure. PMID- 19298438 TI - First-trimester increase in oxidative stress and risk of small-for-gestational age fetus. AB - OBJECTIVE: Investigation of increased oxidative stress in early pregnancy and association with an increased risk of small-for-gestational-age (SGA) fetus. DESIGN: Longitudinal case-control study. SETTING: University Hospitals of Leicester NHS Trust, Leicester, UK. POPULATION: Low-risk pregnant women with no current or pre-existing medical illness were recruited at a large teaching hospital from 2004 to 2006. METHODS: Recruitment performed at the time of the dating ultrasound scan (12+/-2 weeks of gestation). Spot urine samples collected at 12+/-2 and 28+/-2 weeks of gestation were analysed for 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodG) by liquid chromatography with tandem mass spectrometry). SGA was defined as birthweight <10th centile based on customised centile calculator (www.gestation.net). This identified the cases (n=55), whereas controls (n=55) were mothers whose babies were appropriate for gestational age (AGA, birthweight 10th-90th centile). Statistical analysis was performed using GraphPad Prism v.5. The relationship between maternal urinary 8-oxodG at different gestations and customised SGA was investigated by nonparametric tests. MAIN OUTCOME MEASURES: Customised SGA and AGA pregnancies. RESULTS: Urinary 8 oxodG concentrations were significantly increased in pregnancies with subsequent SGA compared with concentrations in normal pregnancies; 12 weeks: 2.8 (interquartile range [IQR] 1.96-3.67) versus 2.2 (IQR 1.26-3.28) pmol 8 oxodG/micromol creatinine (P=0.0007); 28 weeks: 2.21 (IQR 1.67-3.14) versus 1.68 (IQR 1.16-2.82) pmol 8-oxodG/micromol creatinine (P<0.0002). Concentrations decreased significantly between week 12 and 28 (P=0.04 and P=0.02 for controls and cases). CONCLUSIONS: In this study, urinary 8-oxodG at 12 and 28 weeks were elevated in SGA compared with AGA pregnancies. This may reflect early placental changes predating clinical features of SGA. PMID- 19298439 TI - Research priorities in urinary incontinence: results from citizens' juries. AB - OBJECTIVE: The objective of this study was to elicit research ideas, priorities and outcome measures from women who suffer from urinary incontinence. DESIGN: Citizens' juries gather participants together for a combination of education and deliberation on a specific topic. The juries were held in November 2007. SETTING: Women living in the community with urinary incontinence. SAMPLE: Purposively selected responders to public advertisements in Dunedin, New Zealand, were grouped into two juries dependent on whether stress urinary incontinence or urge urinary incontinence was the predominant problem. METHODS: The juries had a day of education in incontinence and research-related issues, with a half day for deliberation and reporting. MAIN OUTCOME MEASURES: Ideas for areas where research may help the lives of women with incontinence in priority order and how best to measure the outcome of that research. RESULTS: The juries identified five main areas for incontinence research with only minor differences depending on which jury. These are, in priority order, making seeking help easier, making day-to-day life more manageable, finding out total costs, more knowledge about causes and the effects of lifestyle. Quality of life was considered by far the most important outcome. CONCLUSIONS: Citizens' juries are able to come up with ideas for research and prioritise these. Research in the areas suggested would be likely to improve the lives of women with urinary incontinence and may lead to a different mix of research projects than is currently the case. PMID- 19298440 TI - Measuring outcomes of importance to women with stress urinary incontinence. AB - OBJECTIVE: To provide evidence on outcomes of importance to women who have stress urinary incontinence (SUI). The secondary aim was to identify additional outcomes that ought to be collected in future primary studies or in systematic reviews of the literature. DESIGN: Questionnaire survey of a cohort of women with SUI. SETTING: UK. SAMPLE: A total of 188 women with SUI. METHODS: Areas of importance to women who suffer from SUI were assessed using a patient generated index (PGI). In addition to the PGI, the questionnaire included the King's Health Questionnaire (KHQ) and the EuroQol-5D (EQ-5D). MAIN OUTCOME MEASURES: PGI, EQ-5D and the KHQ. RESULTS: In total, 38 different areas were reported by respondents on the PGI. PGI and EQ-5D scores were positively correlated and significant. Correlations between the seven domains of the KHQ and PGI were all negative, but only two were statistically significant: personal relationships and severity measures. CONCLUSIONS: The PGI succeeded in capturing a diverse range of outcomes of importance to women suffering with SUI. Given the limited correlation between the KHQ domains and the PGI and, in addition, that the areas mentioned in the PGI were not found to map well to the EQ-5D, the PGI in this instance may be capturing concerns of women who suffer from SUI, which are not captured by quality-of-life measures such as the EQ-5D. PMID- 19298441 TI - Morbidity outcomes of 78,577 hysterectomies for benign reasons over 23 years. PMID- 19298443 TI - Cerebral palsy and restricted growth status at birth: population-based case control study. PMID- 19298444 TI - Maternal mortality in sub-Saharan Africa: the contribution of ineffective blood transfusion services. PMID- 19298446 TI - Pregnancy physiology. PMID- 19298449 TI - Histidine-tryptophan-ketoglutarate (HTK) is associated with reduced graft survival of deceased donor kidney transplants. AB - Single-center studies have reported equivalent outcomes of kidney allografts recovered with histidine-tryptophan-ketoglutarate (HTK) or University of Wisconsin (UW) solution. However, these studies were likely underpowered and often unadjusted, and multicenter studies have suggested HTK preservation might increase delayed graft function (DGF) and reduce graft survival of renal allografts. To further inform clinical practice, we analyzed the United Network for Organ Sharing (UNOS) database of deceased donor kidney transplants performed from July 2004 to February 2008 to determine if HTK (n = 5728) versus UW (n = 15 898) preservation impacted DGF or death-censored graft survival. On adjusted analyses, HTK preservation had no effect on DGF (odds ratio [OR] 0.99, p = 0.7) but was associated with an increased risk of death-censored graft loss (hazard ratio [HR] 1.20, p = 0.008). The detrimental effect of HTK was a relatively late one, with a strong association between HTK and subsequent graft loss in those surviving beyond 12 months (HR 1.43, p = 0.007). Interestingly, a much stronger effect was seen in African-American recipients (HR 1.55, p = 0.024) than in Caucasian recipients (HR 1.18, p = 0.5). Given recent studies that also demonstrate that HTK preservation reduces liver and pancreas allograft survival, we suggest that the use of HTK for abdominal organ recovery should be reconsidered. PMID- 19298450 TI - Early hepatic artery thrombosis after liver transplantation: a systematic review of the incidence, outcome and risk factors. AB - To clarify inconsistencies in the literature we performed a systematic review to identify the incidence, risk factors and outcome of early hepatic artery thrombosis (eHAT) after liver transplantation. We searched studies identified from databases (MEDLINE, EMBASE, Science Citation Index) and references of identified studies. Seventy-one studies out of 999 screened abstracts were eligible for this systematic review. The incidence of eHAT was 4.4% (843/21, 822); in children 8.3% and 2.9% in adults (p < 0.001). Doppler ultrasound screening (DUS) protocols varied from 'no routine' to 'three times a day.' The median time to detection was at day seven. The overall retransplantation rate was 53.1% and was higher in children (61.9%) than in adults (50%, p < 0.03). The overall mortality rate of patients with eHAT was 33.3% (range: 0-80%). Mortality in adults (34.3%) was higher than in children (25%, p < 0.03). The reported risk factors for eHAT were, cytomegalovirus mismatch (seropositive donor liver in seronegative recipient), retransplantation, arterial conduits, prolonged operation time, low recipient weight, variant arterial anatomy, and low volume transplantation centers. eHAT is associated with significant graft loss and mortality. Uniform definitions of eHAT and uniform treatment modalities are obligatory to confirm these results and to obtain a better understanding of this disastrous complication. PMID- 19298451 TI - Treatment-dependent loss of polyfunctional CD8+ T-cell responses in HIV-infected kidney transplant recipients is associated with herpesvirus reactivation. AB - Antiretroviral-therapy has dramatically changed the course of HIV infection and HIV-infected (HIV(+)) individuals are becoming more frequently eligible for solid organ transplantation. However, only scarce data are available on how immunosuppressive (IS) strategies relate to transplantation outcome and immune function. We determined the impact of transplantation and immune-depleting treatment on CD4+ T-cell counts, HIV-, EBV-, and Cytomegalovirus (CMV)-viral loads and virus-specific T-cell immunity in a 1-year prospective cohort of 27 HIV(+) kidney transplant recipients. While the results show an increasing breadth and magnitude of the herpesvirus-specific cytotoxic T-cell (CTL) response over time, they also revealed a significant depletion of polyfunctional virus-specific CTL in individuals receiving thymoglobulin as a lymphocyte-depleting treatment. The disappearance of polyfunctional CTL was accompanied by virologic EBV reactivation events, directly linking the absence of specific polyfunctional CTL to viral reactivation. The data provide first insights into the immune-reserve in HIV+ infected transplant recipients and highlight new immunological effects of thymoglobulin treatment. Long-term studies will be needed to assess the clinical risk associated with thymoglobulin treatment, in particular with regards to EBV associated lymphoproliferative diseases. PMID- 19298452 TI - Upregulation of TNF receptor type 2 in human and experimental renal allograft rejection. AB - An important role of TNF interacting with TNFR2 has been shown in different models of ischemic, nephrotoxic and immune-mediated renal injury. To systematically evaluate the expression of TNFR2 in renal allograft rejection, we investigated human renal allograft biopsies and, in addition, established an experimental transplantation model in rats to verify the human data under standardized conditions. The expression of TNFR2 was analyzed in 96 human renal allograft biopsies with different disease entities. In a 6-day and a 28-day experimental protocol, TNFR2 was examined in kidney specimens and in the urine of control, uni-nephrectomized and transplanted rats +/- cyclosporine treatment (n = 114). In human biopsies and in rat allografts on day 6 with acute allograft rejection, significantly elevated expression of TNFR2 was observed in tubular epithelial cells, podocytes, B cells and monocytes/macrophages. The expression level was associated with renal function. The TNFR2 expression level at day 28 was significantly lower compared to day 6. TNFR2 is markedly upregulated both in human and experimental acute renal allograft rejection. Our data are robust and consistent between different species, suggesting a role for TNFR2 in the early course of rejection. PMID- 19298453 TI - Immediate retransplantation for pancreas allograft thrombosis. AB - Early pancreas allograft failure most commonly results from thrombosis and requires immediate allograft pancreatectomy. Optimal timing for retransplantation remains undefined. Immediate retransplantation facilitates reuse of the same anatomic site before extensive adhesions have formed. Some studies suggest that early retransplantation is associated with a higher incidence of graft loss. This study is a retrospective review of immediate pancreas retransplants performed at a single center. All cases of pancreas allograft loss within 2 weeks were examined. Of 228 pancreas transplants, 12 grafts were lost within 2 weeks of surgery. Eleven of these underwent allograft pancreatectomy for thrombosis. One suffered anoxic brain injury and was not a retransplantation candidate, one was retransplanted at 3.5 months and nine patients underwent retransplantation 1-16 days following the original transplant. Of the nine early retransplants, one pancreas was lost to heparin-induced thrombocytopenia, one recipient died with function at 2.9 years and the other grafts continue to function at 76-1137 days (mean 572 days). One-year graft survival for early retransplantation was 89% compared to 91% for all pancreas transplants at our center. Immediate retransplantation following pancreatic graft thrombosis restores durable allograft function with outcomes comparable to first-time pancreas transplantation. PMID- 19298454 TI - The fou2 mutation in the major vacuolar cation channel TPC1 confers tolerance to inhibitory luminal calcium. AB - The SV channel encoded by the TPC1 gene represents a Ca(2+)- and voltage dependent vacuolar cation channel. Point mutation D454N within TPC1, named fou2 for fatty acid oxygenation upregulated 2, results in increased synthesis of the stress hormone jasmonate. As wounding causes Ca2+ signals and cytosolic Ca2+ is required for SV channel function, we here studied the Ca(2+)-dependent properties of this major vacuolar cation channel with Arabidopsis thaliana mesophyll vacuoles. In patch clamp measurements, wild-type and fou2 SV channels did not exhibit differences in cytosolic Ca2+ sensitivity and Ca2+ impermeability. K+ fluxes through wild-type TPC1 were reduced or even completely faded away when vacuolar Ca2+ reached the 0.1-mm level. The fou2 protein under these conditions, however, remained active. Thus, D454N seems to be part of a luminal Ca2+ recognition site. Thereby the SV channel mutant gains tolerance towards elevated luminal Ca2+. A three-fold higher vacuolar Ca/K ratio in the fou2 mutant relative to wild-type plants seems to indicate that fou2 can accumulate higher levels of vacuolar Ca(2+) before SV channel activity vanishes and K(+) homeostasis is impaired. In response to wounding fou2 plants might thus elicit strong vacuole derived cytosolic Ca2+ signals resulting in overproduction of jasmonate. PMID- 19298456 TI - Molecular structure in peripheral dog breeds: Portuguese native breeds as a case study. AB - Genetic variability in purebred dogs is known to be highly structured, with differences among breeds accounting for approximately 30% of the genetic variation. However, analysis of the genetic structure in non-cosmopolitan breeds and local populations is still limited. Nine Portuguese native dog breeds, and other peripheral dog populations (five) with regional affinities, were characterized using 16 microsatellites and 225 amplified fragment length polymorphism (AFLP) markers, and the pattern of genetic differentiation was investigated. Although the level of breed differentiation detected is below that of other dog breeds, there is in most cases a correlation between breed affiliation and molecular structure. AFLP markers and Bayesian clustering methods allowed an average of 73.1% of individuals to be correctly assigned to source populations, providing robust genotypic assessment of breed affiliation. A geographical genetic structure was also detected, which suggests a limited influence of African dogs on the Iberian breeds. The sampling effect on the estimation of population structure was evaluated and there was a 2.2% decrease in genetic differentiation among breeds when working animals were included. Genetic diversity of stray dogs was also assessed and there is no evidence that they pose a threat to the preservation of the gene pool of native dog breeds. PMID- 19298457 TI - The positive and negative health effects of alcohol- and the public health implications. AB - In this paper, the negative and the positive effects of alcohol on health are reviewed. It is first of all established facts that a high alcohol intake implies an increased risk of a large number of health outcomes, such as dementia, breast cancer, colorectal cancer, cirrhosis, upper digestive tract cancer and alcohol dependency. Second, it is justified that alcohol has beneficial effects for some individuals, especially with regard to prevention of thrombosis of the heart. The public health relevance of these results is considered. The sensible drinking limits, used in both the UK and Denmark, of a maximum of 21 drinks per week for men and 14 drinks per week for women seem valid. A broader public health message of the beneficial effects of alcohol does not seem to be of interest in Western societies, where only a very small fraction of the population are non drinkers and may have very good reasons therefore. PMID- 19298458 TI - Infectious agents and lymphoma development: molecular and clinical aspects. AB - This review is focused on the role of infectious agents in the development of some lymphoma entities. Associations involving bacterial infections mostly regard marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT)-type. Some paradigmatic examples of these associations include the Helicobacter pylori related gastric MALT lymphoma and the more recently reported links between Chlamydophila psittaci and ocular adnexal MALT lymphomas and Borrelia burgdorferi and cutaneous MALT lymphomas. The well-documented association between Epstein Barr virus infection and related lymphoproliferative disorders are analysed as an example of lymphotropic virus with tumourigenic activity. Molecular, biological and clinical features as well as therapeutic implications of these associations are analysed and future perspectives in this field are discussed. PMID- 19298459 TI - Free fatty acid kinetics during long-term treatment with pioglitazone added to sulfonylurea or metformin in Type 2 diabetes. AB - BACKGROUND: Free fatty acids (FFAs) are linked to impaired insulin action, but their role in mediating long-term insulin sensitization during diabetes treatment is unclear. OBJECTIVES: To examine the effect of pioglitazone addition to existing therapy on FFA dynamics and insulin action. DESIGN: Two 2-year, randomized, parallel-group, double-blind, double-dummy, clinical trials. SETTING: One hundred and seventy-one centres in Europe, Australia and Canada. SUBJECTS: Male and female patients with Type 2 diabetes inadequately managed with metformin or sulfonylurea. INTERVENTIONS: Patients were randomized to pioglitazone (15-45 mg day(-1); n=319) or metformin (850-2550 mg day(-1); n=320) as add-on therapy to gliclazide or pioglitazone (n=317) versus gliclazide (80-320 mg day(-1); n=313) as add-on therapy to metformin. OUTCOME MEASURE: Plasma FFA profiles during oral glucose tolerance tests in selected centres before and during treatment (n=588). RESULTS: At Week 104, pioglitazone treatment decreased fasting FFAs by 0.08 mmol L(-1) when added to sulfonylurea and by 0.11 mmol L(-1) when added to metformin versus the respective sulfonylurea + metformin groups (0.03 mmol L(-1), P=0.05 and 0.04 mmol L(-1), P<0.05), and this was accompanied by significant improvements in fasting adipose tissue insulin sensitivity. Changes in postchallenge FFAs were similar between groups and not related to changes in liver transaminases, insulin action and secretion. However, the sensitivity of FFA to insulin was affected by treatment (P<0.001) and visit (P<0.05). Insulin sensitivity of FFA rose when pioglitazone was added to sulfonylurea (P<0.05), but decreased for gliclazide + metformin (P<0.05). CONCLUSION: Long-term improvements in adipose tissue insulin sensitivity and reduction in fasting FFAs with pioglitazone may help to reduce lipotoxicity in Type 2 diabetes. PMID- 19298461 TI - Analyzing salvia divinorum and its active ingredient salvinorin a utilizing thin layer chromatography and gas chromatography/mass spectrometry. AB - In recent years, Salvia divinorum has become a major focus by state legislatures throughout the United States looking to prohibit the sale of the psychoactive plant. After researching testing procedures presented in the literature and those employed by crime laboratories throughout the country, it was decided that thin layer chromatography (TLC) and gas chromatography/mass spectrometry (GC/MS) were the methods to use to analyze plant material for salvinorin A. With TLC, salvinorin A was detected from extracted plant material and was easily distinguishable from 13 other Salvia species as well as Cannabis sativa L. (marijuana). When using GC/MS, salvinorin A was best extracted from plant material with chloroform at ambient temperature when using a nonpolar solvent and acetone at ambient temperature when using a polar solvent. By utilizing these techniques, criminalists are now able to confirm the presence of salvinorin A in a submitted plant material suspected to be Salvia divinorum. PMID- 19298462 TI - Fatal venous air embolism following intravenous infusion. AB - Venous air embolism has been reported as a complication of invasive diagnostic and therapeutic procedures or accidental trauma. Little is known about the incidence of air embolism after minimal intravenous manipulations such as the insertion of a peripheral intravenous cannula. Only when large amounts of air sufficient enough to block the cardiovascular system enter, the patient develops symptoms and signs of severe neurological injury, cardiovascular collapse, or death. The dead body of a 14-year-old boy was brought for postmortem examination with allegations of death from negligence during treatment. He was treated for pain in the abdomen in a hospital by attendants in telephonic consultation with a medical practitioner. Following intravenous infusions, the boy died suddenly in respiratory distress. Gross findings indicated the death to be from venous air embolism. Chemical analysis, histopathology, and microbiology ruled out other causes of death. Dilemmas of the case with difficulties in diagnosis are being presented herewith. PMID- 19298463 TI - Forensic characteristics of suicide by electrocution in Bulgaria. AB - Suicides by electrocution (SE) are relatively rare and insufficiently studied. The purpose of this study is to present a more detailed picture of SE in Bulgaria. Forensic medical files of 59 SE cases were examined. SPSS 11.0 software was applied to produce the descriptive statistical analyses. Values of p < 0.05 were assumed to be statistically significant. We studied 59 SE cases, which account for 6.24% of all deaths by electrocution and 0.09% of all forensic autopsies. The average age of victims was 45.19 years old. Males prevailed: 91.53%. Children below 18 years old accounted for 3.39%. 42.37% of the victims applied low voltage (<220 V), and 33.99% applied high voltage (>220 V). The preferred method of SE was by direct contact victim/electrical cable: 47.45%. The summer months June through September account for 49.15% of all cases. PMID- 19298464 TI - Histological determination of the human origin of bone fragments. AB - A frequently encountered task in the forensic scenario is verification of the human origin of severely degraded fragments of bone. In these cases histological methods which consider osteon size and morphology can prove to be useful. The authors in the present study verify the applicability of published algorithms to flat and subadult bones from human, dog, cat, cow, rabbit, sheep, pig, chicken, quail, and turkey samples. Metric analysis was performed on 2031 Haversian canals. Analyses carried out on human samples confirmed a success rate of around 70% on long adult bones; however the percentage of wrong answers was particularly high in the case of newborns and older subadults as well as on flat bones in general. Results therefore suggest that such regression equations should be limited only to bone fragments from long adult bones. PMID- 19298465 TI - Extraction and Refinement Strategy for detection of autism in 18-month-olds: a guarantee of higher sensitivity and specificity in the process of mass screening. AB - BACKGROUND: For early detection of autism, it is difficult to maintain an efficient level of sensitivity and specificity based on observational data from a single screening. The Extraction and Refinement (E&R) Strategy utilizes a public children's health surveillance program to produce maximum efficacy in early detection of autism. In the extraction stage, all cases at risk of childhood problems, including developmental abnormality, are identified; in the refinement stage, cases without problems are excluded, leaving only cases with conclusive diagnoses. METHODS: The city of Yokohama, Japan, conducts a routine child health surveillance program for children at 18 months in which specialized public health nurses administer YACHT-18 (Young Autism and other developmental disorders CHeckup Tool), a screening instrument to identify children at risk for developmental disorders. Children who screen positive undergo further observation, and those without disorders are subsequently excluded. To study the efficacy of early detection procedures for developmental disorders, including autism, 2,814 children born in 1988, examined at 18 months of age, and not already receiving treatment for diseases or disorders were selected. RESULTS: In the extraction stage, 402 (14.3%) children were identified for follow-up. In the refinement stage, 19 (.7%) of these were referred to the Yokohama Rehabilitation Center and diagnosed with developmental disorders. The extraction stage produced four false negatives, bringing total diagnoses of developmental disorders to 23 (.8%) - including 5 with autistic disorder and 9 with pervasive developmental disorder - not otherwise specified (PDDNOS). Sensitivity was 60% for autistic disorder and 82.6% for developmental disorders. Specificity for developmental disorders rose to 100% with the E&R Strategy. Picture cards used in YACHT-18 provided a finer screen that excluded some false positive cases. CONCLUSIONS: An extraction and refinement methodology utilizing child health surveillance programs achieve high efficacy for early detection of autism. PMID- 19298466 TI - Visual orienting in the early broader autism phenotype: disengagement and facilitation. AB - BACKGROUND: Recent studies of infant siblings of children diagnosed with autism have allowed for a prospective approach to examine the emergence of symptoms and revealed behavioral differences in the broader autism phenotype within the early years. In the current study we focused on a set of functions associated with visual attention, previously reported to be atypical in autism. METHOD: We compared performance of a group of 9-10-month-old infant siblings of children with autism to a control group with no family history of autism on the 'gap overlap task', which measures the cost of disengaging from a central stimulus in order to fixate a peripheral one. Two measures were derived on the basis of infants' saccadic reaction times. The first is the Disengagement effect, which measures the efficiency of disengaging from a central stimulus to orient to a peripheral one. The second was a Facilitation effect, which arises when the infant is cued by a temporal gap preceding the onset of the peripheral stimulus, and would orient faster after its onset. RESULTS AND CONCLUSION: Infant siblings of children with autism showed longer Disengagement latencies as well as less Facilitation relative to the control group. The findings are discussed in relation to how differences in visual attention may relate to characteristics observed in autism and the broader phenotype. PMID- 19298467 TI - Situational and generalised conduct problems and later life outcomes: evidence from a New Zealand birth cohort. AB - BACKGROUND: There is considerable evidence suggesting that many children show conduct problems that are specific to a given context (home; school). What is less well understood is the extent to which children with situation-specific conduct problems show similar outcomes to those with generalised conduct problems. METHODS: Data were gathered as part of the Christchurch Health and Development Study, a 25-year longitudinal study of a birth cohort of New Zealand children. Information was obtained on: (a) conduct problems during the period 7-9 years; (b) criminal offending during the period 16-25 years; (c) measures of DSM IV mental disorders and suicidal behaviour over the interval 16-25 years; (d) measures of DSM-IV substance dependence over the interval 16-25 years; and (e) measures of relationship, pregnancy, and parenthood outcomes during the period 16 25 years. RESULTS: Latent-class modelling suggested three distinct groups of children with conduct problems: those with mother reports; those with teacher reports; and those with both mother and teacher reports. Both situation-specific and generalised conduct disorder were associated with increased risk of criminal offending, mental health disorders, substance dependence, and relationship and parenthood issues in late adolescence and early adulthood. CONCLUSIONS: There is a need for recognition of the significance of situation-specific conduct problems in both developmental theory and in the treatment of childhood conduct disorders. A focus only on those children with generalised conduct problems is likely to overlook the features and needs of children whose conduct problems are confined to a specific context. PMID- 19298468 TI - Conscientiousness as a mediator of the association between masculinized finger length ratios and attention-deficit/hyperactivity disorder (ADHD). AB - BACKGROUND: One often-overlooked biological risk factor that may help explain sex biased prevalence rates in psychopathology is sex hormones. Personality traits, which also show sex differences, may mediate relations between biological risk factors like hormones and childhood psychopathology such as ADHD (or, alternatively, be independent risk factors). METHODS: Three hundred and twelve children/adolescents (178 boys, 134 girls) between the ages of 8 and 17 completed a comprehensive, multistage, clinical diagnostic procedure; 168 children were diagnosed with ADHD and 144 were classified as non-ADHD comparison controls. Primary caregivers completed the California Q-sort in order to provide a measure of conscientiousness. Finger-length ratios (specifically right 2D:4D) served as a proxy of prenatal testosterone exposure (relative to estrogen). RESULTS: Lower levels of conscientiousness statistically mediated the relationship between more masculine right 2D:4D (i.e., increased prenatal testosterone exposure) and increased ADHD inattentive symptoms. CONCLUSION: More masculinized finger-length ratios show associations with ADHD symptoms, possibly acting through the trait mechanism of conscientiousness. PMID- 19298469 TI - Directional effects between rapid auditory processing and phonological awareness in children. AB - BACKGROUND: Deficient rapid auditory processing (RAP) has been associated with early language impairment and dyslexia. Using an auditory masking paradigm, children with language disabilities perform selectively worse than controls at detecting a tone in a backward masking (BM) condition (tone followed by white noise) compared to a forward masking (FM) condition (tone preceded by white noise). Tallal's (1980) auditory processing hypothesis posits that abnormal RAP leads to reduced (or impaired) phonological awareness (PA), resulting in reading and language difficulties. Alternative theories suggest that impaired PA may have more of a top-down effect on auditory processing. METHODS: The current study examines RAP in children tested at two time points, average age 5.6 and average age 8.3, in order to test causal relations between RAP and PA in a path analysis. Additional hierarchical regressions examine how well RAP predicts reading ability when accounting for PA and vocabulary. RESULTS: The path analysis indicates a top down effect, such that PA has a larger impact on BM over time than the reverse. Regressions indicate no direct impact of RAP on reading ability. DISCUSSION: The path analysis provides evidence against the auditory processing hypothesis and instead suggests that between the ages of 5 and 8 it is variability in early phonological representations that predicts subsequent lower-level rapid auditory processing. PMID- 19298470 TI - Familial associations of intense preoccupations, an empirical factor of the restricted, repetitive behaviors and interests domain of autism. AB - BACKGROUND: Clinical heterogeneity of autism likely hinders efforts to find genes associated with this complex psychiatric disorder. Some studies have produced promising results by restricting the sample according to the expression of specific familial factors or components of autism. Previous factor analyses of the restricted, repetitive behaviors and interest (RRBI) domain of autism have consistently identified a two-factor model that explains a moderate amount of variance. The identification of additional factors may explain more variance in the RRBI domain and provide an additional component of autism that may help in the identification of underlying genetic association. METHODS: We conducted factor analyses of RRBI symptoms with a sample that included verbal subjects meeting full criteria for autism aged 5 to 22 years (n = 245). Among affected sibling pairs (n = 126) we examined the familial aggregation of the identified factors. We also examined the associations of the factors with autism-related personality traits in fathers and mothers (n = 50). RESULTS: The previously identified two-factor model - insistence on sameness (IS) and repetitive stereotypic motor behaviors (RSMB) - was replicated in our sample. Next, a second factor analysis that included the item for verbal rituals resulted in a four factor model - IS, 'simple' RSMB, 'complex' RSMB, and a fourth factor including symptoms associated with intense preoccupations (IP). Of these four, both IS and IP were significantly familial among affected siblings, but only IP was significantly correlated with the broader autism phenotype traits of rigidity and aloofness in fathers. CONCLUSIONS: The results support previous evidence for the IS factor, its familiality, and the identification of IP as an additional strong candidate trait for genetic studies of autism. PMID- 19298471 TI - Self-referenced memory, social cognition, and symptom presentation in autism. AB - BACKGROUND: We examined performance on a self-referenced memory (SRM) task for higher-functioning children with autism (HFA) and a matched comparison group. SRM performance was examined in relation to symptom severity and social cognitive tests of mentalizing. METHOD: Sixty-two children (31 HFA, 31 comparison; 8-16 years) completed a SRM task in which they read a list of words and decided whether the word described something about them, something about Harry Potter, or contained a certain number of letters. They then identified words that were familiar from a longer list. Dependent measures were memory performance (d') in each of the three encoding conditions as well as a self-memory bias score (d' self-d' other). Children completed The Strange Stories Task and The Children's Eyes Test as measures of social cognition. Parents completed the SCQ and ASSQ as measures of symptom severity. RESULTS: Children in the comparison sample showed the standard SRM effect in which they recognized significantly more self referenced words relative to words in the other-referenced and letter conditions. In contrast, HFA children showed comparable rates of recognition for self- and other-referenced words. For all children, SRM performance improved with age and enhanced SRM performance was related to lower levels of social problems. These associations were not accounted for by performance on the mentalizing tasks. CONCLUSIONS: Children with HFA did not show the standard enhanced processing of self- vs. other-relevant information. Individual differences in the tendency to preferentially process self-relevant information may be associated with social cognitive processes that serve to modify the expression of social symptoms in children with autism. PMID- 19298472 TI - An observational study of the interactions of socially withdrawn/anxious early adolescents and their friends. AB - BACKGROUND: The friendships of socially withdrawn/anxious children and early adolescents have been found to lack critical rewarding qualities. Observational research may help elucidate the obstacles they face in forming and maintaining high-quality friendships with sociable peers. METHOD: We observed the interactions of 38 socially withdrawn early adolescents with their friends and compared them to a community control group. RESULTS: In negotiating the sharing of an object, the socially withdrawn, anxious group was more passive than controls. The socially withdrawn, anxious participants engaged less actively in a fast-paced game involving miniature cars. While completing a quiet drawing task, the socially anxious, withdrawn participants tended to refrain from comparing their work to that of their friends. In all three of our closed-field situations, the socially withdrawn, anxious participants displayed relatively neutral affect in comparison with the control group. CONCLUSIONS: These results suggest that the social withdrawal and social anxiety of children with social phobia are very evident even within the confines of their close friendships. Therefore, therapeutic interventions at the level of the dyad may be indicated. PMID- 19298473 TI - Do individuals with autism spectrum disorders infer traits from behavior? AB - BACKGROUND: Traits and mental states are considered to be inter-related parts of theory of mind. Attribution research demonstrates the influential role played by traits in social cognition. However, there has been little investigation into how individuals with autism spectrum disorders (ASD) understand traits. METHOD: The ability of individuals with ASD to infer traits from descriptions of behavior was investigated by asking participants to read trait-implying sentences and then to choose one of two words that best related to the sentence. RESULTS: In Experiment 1, individuals with ASD performed similarly to matched controls in being faster at choosing the trait in comparison to the semantic associate of one of the words in the sentence. The results from Experiments 1 and 2 provided converging evidence in suggesting that inferring traits from textual descriptions of behavior occurs with relatively little effort. The results of Experiment 3 suggested that making trait inferences took priority over inferring actions or making semantic connections between words. CONCLUSIONS: Individuals with ASD infer traits from descriptions of behavior effortlessly and spontaneously. The possibility of trait inference being a spared socio-cognitive function in autism is discussed. PMID- 19298474 TI - Emanuel Miller Lecture: Attachment insecurity, disinhibited attachment, and attachment disorders: where do research findings leave the concepts? AB - BACKGROUND: Despite the evidence on anomalous attachment patterns, there has been a tendency to interpret most of these as reflecting differences in security/insecurity. METHODS: Empirical research findings are reviewed in relation to attachment/insecurity as evident in both infancy and later childhood, disorganised attachment, inhibited attachment disorder, and disinhibited attachment disorder. FINDINGS: Substantial differences are found in the correlates and meaning of these different features, as well as in the patterns associated with conditions such as autism, psychopathy, and Williams syndrome. CONCLUSIONS: It is seriously misleading to view all of these patterns through the lens of security/insecurity. This heterogeneity in social relationship features necessarily has implications for the assessment measures for social relationships that need to be used. PMID- 19298475 TI - Different neurocognitive functions regulating physical aggression and hyperactivity in early childhood. AB - BACKGROUND: There are strong parallels between early childhood and adolescent behavior problems. However, we do not know if behavioral symptoms associate with neurocognitive processes in very young children as they do in older children. METHODS: We studied a population-based birth cohort of children (N = 1,950) whose developmental trajectories of physical aggression and hyperactivity were assessed between the ages of 17 and 41 months. We measured the following neurocognitive abilities at 41 months of age: Receptive vocabulary, visuospatial organization, and short-term memory. RESULTS: After controlling for other neurocognitive abilities, frequent physical aggression was related specifically to receptive vocabulary deficits (p < .0001) while frequent hyperactivity was related specifically to deficits of visuospatial organization (p < .0001). The pattern of associations was robust despite controls for socioeconomic and perinatal status. CONCLUSIONS: The different neurocognitive correlates of physical aggression and hyperactivity problems observed during adolescence are apparent in early childhood. Whereas physical aggression problems are associated with language deficits, hyperactivity problems are related to non-verbal deficits. PMID- 19298476 TI - Environmental influences on the longitudinal covariance of expressive vocabulary: measuring the home literacy environment in a genetically sensitive design. AB - BACKGROUND: Despite the well-replicated relationship between the home literacy environment and expressive vocabulary, few studies have examined the extent to which the home literacy environment is associated with the development of early vocabulary ability in the context of genetic influences. This study examined the influence of the home literacy environment on the longitudinal covariance of expressive vocabulary within a genetically sensitive design. METHODS: Participants were drawn from the Western Reserve Reading Project, a longitudinal twin project of 314 twin pairs based in Ohio. Twins were assessed via three annual home visits during early elementary school; expressive vocabulary was measured via the Boston Naming Test (BNT), and the Home Literacy Environment (HLE) was assessed using mothers' report. RESULTS: The heritability of the BNT was moderate and significant at each measurement occasion, h(2) = .29-.49, as were the estimates of the shared environment, c(2) = .27-.39. HLE accounted for between 6-10% of the total variance in each year of vocabulary assessment. Furthermore, 7-9% of the total variance of the stability over time in BNT was accounted for by covariance in the home literacy environment. CONCLUSIONS: These results indicate that aspects of the home literacy environment, as reported by mothers, account for some of the shared environmental variance associated with expressive vocabulary in school aged children. PMID- 19298477 TI - Eye-movement patterns are associated with communicative competence in autistic spectrum disorders. AB - BACKGROUND: Investigations using eye-tracking have reported reduced fixations to salient social cues such as eyes when participants with autism spectrum disorders (ASD) view social scenes. However, these studies have not distinguished different cognitive phenotypes. METHODS: The eye-movements of 28 teenagers with ASD and 18 typically developing peers were recorded as they watched videos of peers interacting in familiar situations. Within ASD, we contrasted the viewing patterns of those with and without language impairments. The proportion of time spent viewing eyes, mouths and other scene details was calculated, as was latency of first fixation to eyes. Finally, the association between viewing patterns and social-communicative competence was measured. RESULTS: Individuals with ASD and age-appropriate language abilities spent significantly less time viewing eyes and were slower to fixate the eyes than typically developing peers. In contrast, there were no differences in viewing patterns between those with language impairments and typically developing peers. Eye-movement patterns were not associated with social outcomes for either language phenotype. However, increased fixations to the mouth were associated with greater communicative competence across the autistic spectrum. CONCLUSIONS: Attention to both eyes and mouths is important for language development and communicative competence. Differences in fixation time to eyes may not be sufficient to disrupt social competence in daily interactions. A multiple cognitive deficit model of ASD, incorporating different language phenotypes, is advocated. PMID- 19298478 TI - Is prenatal alcohol exposure related to inattention and hyperactivity symptoms in children? Disentangling the effects of social adversity. AB - BACKGROUND: Studies concerning whether exposure to low levels of maternal alcohol consumption during fetal development is related to child inattention and hyperactivity symptoms have shown conflicting results. We examine the contribution of covariates related to social adversity to resolve some inconsistencies in the extant research by conducting parallel analyses of three cohorts with varying alcohol consumption and attitudes towards alcohol use. METHODS: We compare three population-based pregnancy-offspring cohorts within the Nordic Network on ADHD from Denmark and Finland. Prenatal data were gathered via self-report during pregnancy and birth outcomes were abstracted from medical charts. A total of 21,678 reports concerning inattention and hyperactivity symptoms in children were available from the Strengths and Difficulties Questionnaire or the Rutter Scale completed by parents and/or teachers. RESULTS: Drinking patterns differed cross-nationally. Women who had at least some social adversity (young, low education, or being single) were more likely to drink than those better off in the Finnish cohort, but the opposite was true for the Danish cohorts. Prenatal alcohol exposure was not related to risk for a high inattention hyperactivity symptom score in children across cohorts after adjustment for covariates. In contrast, maternal smoking and social adversity during pregnancy were independently and consistently associated with an increase in risk of child symptoms. CONCLUSIONS: Low doses of alcohol consumption during pregnancy were not related to child inattention/hyperactivity symptoms once social adversity and smoking were taken into account. PMID- 19298479 TI - Rates and types of psychiatric disorders in perinatally human immunodeficiency virus-infected youth and seroreverters. AB - BACKGROUND: The purpose of this study was to examine 1) the prevalence of psychiatric and substance use disorders in perinatally HIV-infected (HIV+) adolescents and 2) the association between HIV infection and these mental health outcomes by comparing HIV+ youths to HIV exposed but uninfected youths (HIV-) from similar communities. METHODS: Data for this paper come from the baseline interview of a longitudinal study of mental health outcomes in 9-16 year old perinatally HIV-exposed youths (61% HIV+) and their caregivers. Three hundred forty youths and their primary adult caregivers were recruited from four medical centers and participated in separate individual interviews. Youth psychiatric disorder was assessed using the caregiver and youth versions of The Diagnostic Interview Schedule for Children (DISC-IV). RESULTS: According to caregiver or youth report, a high percentage of HIV+ and HIV- youths met criteria for a non substance use psychiatric disorder, with significantly higher rates among the HIV+ youths (61% vs. 49%, OR = 1.59; CI = 1.03,2.47; p < .05). The most prevalent diagnoses in both groups were anxiety disorders (46% for total sample) which included social phobia, separation anxiety, agoraphobia, generalized anxiety disorder, panic disorder, obsessive- compulsive disorder, and specific phobias. One quarter of the sample met criteria for a behavioral disorder (ADHD, conduct disorders, and oppositional defiant disorders), with ADHD being most prevalent. HIV+ youths had significantly higher rates of ADHD (OR = 2.45; CI = 1.20, 4.99, p < .05). Only 7% of youths met criteria for a mood disorder and 4% for a substance abuse disorder. Several caregiver variables (caregiver type and HIV status) were also associated with both child HIV status and mental health outcomes. CONCLUSIONS: Our data suggest that HIV+ youths are at high risk for mental health disorders. Further longitudinal research is necessary to understand the etiology, as well as potential protective factors, in order to inform efficacy-based interventions. PMID- 19298480 TI - Comparison of recipient site preparations in epidermal grafting for vitiligo: suction blister and CO2 laser. PMID- 19298481 TI - Quantitative analysis of epithelial papillae in patients with oral lichen planus. AB - BACKGROUND: The oral mucosa is relatively vulnerable to pathological processes, and is often affected by autoimmune and malignant diseases. The oral epithelium is normally non-homogeneous, and joins to the connective tissue through interlocking of its downward projections in the form of papillae. OBJECTIVE: This study aims to conduct a histomorphometric study of the epithelial papillae in patients with oral lichen planus (OLP). MATERIAL AND METHOD: This study was based on 100 cheek mucosa biopsies from patients with OLP (66 white reticular and 34 atrophic-erosive) (13 males and 87 females, with a mean age of 54.95 +/- 13.64 years). A histological and morphometric evaluation was made, based on imaging analysis with MIP software 4.5 for studying the papillary structure in the patients with OLP. RESULTS: The mean epithelial thickness was 227.5 +/- 78.5 microm. The different papillary measures--BLS (distance from basal layer to epithelial surface), DPS (distance from dermal papilla top to epithelial surface), DPW (dermal papilla width), and DPD (interdermal papilla distance between two papillae)--yielded no statistically significant differences with respect to age, sex, smoking and clinical form. However, a significant correlation was observed in relation to papilla width and inflammatory infiltrate (P = 0.031). CONCLUSIONS: The application of this imaging system is useful for measuring variations in epithelial papillary architecture. PMID- 19298482 TI - Fully regressive targetoid haemosiderotic haemangioma. PMID- 19298483 TI - Sparse hairs below frontal hair line are early sign of androgenetic alopecia in men. PMID- 19298484 TI - Photodynamic therapy with methyl aminolevulinate for resistant scalp folliculitis secondary to Demodex infestation. PMID- 19298485 TI - A critical appraisal of evidence-based guidelines for the treatment of psoriasis vulgaris: 'AGREE-ing' on a common base for European evidence-based psoriasis treatment guidelines. AB - BACKGROUND: To further improve the standard of care provided to psoriasis patients in Europe, the European Dermatology Forum and the European Academy of Dermatology and Venereology have initiated a project to develop common European psoriasis guidelines. OBJECTIVE: This paper aims to assess the methodological quality and suitability of evidence-based psoriasis guidelines as a base of common European treatment guidelines. METHODS: A systematic literature search of the National Guidelines Clearinghouse, MEDLINE, EMBASE and GIN database for evidence-based psoriasis treatment guidelines published between 2001 and 2007 was performed and the AGREE instrument was used to assess the methodological quality. RESULTS: Out of 166 hits, we identified three evidence-based guidelines. A Dutch guideline that includes systemic treatments and photo(chemo)therapy, a British guideline that includes biologics only, and a German guideline that includes systemic therapy, photo(chemo)therapy and topical therapy. For the majority of the 23 AGREE items assessed, all three guidelines rated high (3-4/4). The highest score was obtained by the British guidelines with 75 points out of 92, followed by the German guidelines with 74 points and the Dutch guidelines with 73 points. All guidelines showed weaknesses in the field of 'applicability'. CONCLUSION: The three guidelines that could be included rated high enough to be considered 'strongly recommended' and were included to serve as a basis for the new common European guidelines. During the development of the European guidelines, special attention should be paid to meet the requirements of good 'applicability'. PMID- 19298486 TI - Treatment of cutaneous leishmaniasis with intralesional sodium stibogluconate. AB - BACKGROUND: Cutaneous leishmaniasis is endemic in Israel. Leishmania major is the most prevalent species that cause cutaneous leishmaniasis. Current treatment options are limited and there are few investigations in search of alternative ones. OBJECTIVE: This study aims to assess our experience with intralesional sodium stibogluconate (SSG) in the treatment of cutaneous leishmaniasis. METHODS: A retrospective evaluation for all adult cases of cutaneous leishmaniasis treated by intralesional and intravenous SSG (Pentostam, GlaxoSmithKline) between 2004 and 2006 was performed, for cases referred to a tertiary care university affiliated medical centre in Israel. Intralesional SSG was injected at 0.5 mL per lesion (50 mg). Treatment was repeated every 2-3 weeks for a total of 12 weeks. Intravenous SSG was administered at a dose of 20 mg/kg for 10-20 days. RESULTS: Thirty-three cases of cutaneous leishmaniasis were treated with intralesional SSG during the study period. The patients consist of 26 males and 7 females, mostly Israeli military personnel, and there were a total of 93 lesions. Within 3 months from treatment onset, 91% (30/33) had completed healing of the cutaneous lesions after an average of 3 treatments (range 1-6). Side-effects were mild and were mostly pain during injection, with two patients developing mild local site reaction after the injection. CONCLUSIONS: Intralesional SSG treatment is safe, effective and well tolerated with minimal side-effects. PMID- 19298487 TI - Cutaneous sarcoidosis: a histopathological study. AB - BACKGROUND: Sarcoidosis is a granulomatous disease of uncertain aetiology in which the skin is frequently involved. Naked sarcoidal granulomas are the characteristic histological feature in specific lesions of sarcoidosis. OBJECTIVE: This study aims to describe the histological findings in a population of patients with cutaneous sarcoidosis. MATERIALS AND METHODS: This study is a retrospective analysis of 31 biopsies of specific lesions of cutaneous sarcoidosis, corresponding to 30 patients. RESULTS: Typical naked granuloma was found in the majority of cases (71%). In 9 cases (29%), granulomas had a significant number of lymphocytes. Necrosis was found in two cases (6%). Periadnexal distribution (mostly perisudoral) was found in 32% of cases. Interstitial distribution of granulomas was observed in five cases (16%). Foreign material was detected in 13% of cases (without the use of polarized light microscopy). Epidermal changes were found in 55% of cases, with atrophy and parakeratosis being the most frequent alterations. CONCLUSIONS: Although typical naked sarcoid granulomas are the most common features of cutaneous sarcoidosis, the dermatopathologist must be aware of possible atypical findings, which are more common than previously expected, because of the differential diagnosis with other causes of cutaneous granulomas, namely infectious diseases. PMID- 19298488 TI - Pemphigus and drug addiction. PMID- 19298489 TI - New dermoscopic vascular patterns in a case of eccrine poroma. PMID- 19298490 TI - Oral zinc sulphate in recalcitrant multiple viral warts: a pilot study. PMID- 19298491 TI - Annular lichenoid dermatosis of youth. PMID- 19298492 TI - Quality-quantity trade-off of human offspring under adverse environmental conditions. AB - A central paradigm in life-history theory is the trade-off between offspring number and quality. Several studies have investigated this trade-off in humans, but data are inconclusive, perhaps because prosperous socio-cultural factors mask the trade-off. Therefore, we studied 2461 offspring groups in an area under adverse conditions in northern Ghana with high fertility and mortality rates. In a linear mixed model controlling for differences in age and tribe of the mother and socioeconomic status, each additional child in the offspring group resulted in a 2.3% (95% CI 1.9-2.6%, P < 0.001) lower proportional survival of the offspring. Furthermore, we made use of the polygamous population structure and compared offspring of co-wives in 388 households, thus controlling for variation in resources between compounds. Here, offspring survival decreased 2.8% (95% CI 2.3-4.0%, P < 0.001) for each increase in offspring number. We interpret these data as an apparent quality-quantity trade-off in human offspring. PMID- 19298493 TI - The cost of multiple drug resistance in Pseudomonas aeruginosa. AB - The spread of bacterial antibiotic resistance mutations is thought to be constrained by their pleiotropic fitness costs. Here we investigate the fitness costs of resistance in the context of the evolution of multiple drug resistance (MDR), by measuring the cost of acquiring streptomycin resistance mutations (StrepR) in independent strains of the bacterium Pseudomonas aeruginosa carrying different rifampicin resistance (RifR) mutations. In the absence of antibiotics, StrepR mutations are associated with similar fitness costs in different RifR genetic backgrounds. The cost of StrepR mutations is greater in a rifampicin sensitive (RifS) background, directly demonstrating antagonistic epistasis between resistance mutations. In the presence of rifampicin, StrepR mutations have contrasting effects in different RifR backgrounds: StrepR mutations have no detectable costs in some RifR backgrounds and massive fitness costs in others. Our results clearly demonstrate the importance of epistasis and genotype-by environment interactions for the evolution of MDR. PMID- 19298494 TI - Investment in the public good through conditional phenotypes of large effect. AB - We investigate the evolution of an individual's willingness to invest in a public good (what we call, helping) in a patch-structured population with limited natal dispersal. We assume that an individual's decision to invest is informed by its dispersal status: an individual makes one decision given it is native to the patch on which it breeds, and is free to make a different decision given that it is not native to the patch on which it breeds. Unlike previous work, we assume that investment in the public good, and the public good, itself, both have a large effect on individual fecundity. Kin selection analysis reveals that only extreme investment decisions (i.e. 'always invest' or 'never invest') can be evolutionarily stable. Numerical results suggest that the evolutionary instability of the 'never invest' phenotype (what we call, complete nonhelping) implies the evolutionary stability of 'always invest' (what we call, complete helping). In addition, numerical results show that bistability of extreme phenotypes is possible, indicating that the adaptive significance of altruism, in this context, is greater than has been previously recognized. Numerical results are supported by computer simulation, and results, themselves, are briefly discussed in a concluding section. PMID- 19298495 TI - Lifetime reproductive success and longevity of queens in an annual social insect. AB - Although central to understanding life-history evolution, the relationship between lifetime reproductive success and longevity remains uncertain in many organisms. In social insects, no studies have reported estimates of queens' lifetime reproductive success and longevity within populations, despite the importance of understanding how sociality and associated within-group conflict affect life-history traits. To address this issue, we studied two samples of colonies of the annual bumblebee, Bombus terrestris audax, reared from wild caught queens from a single population. In both samples, queens' lifetime reproductive success, measured as either queens' inclusive fitness or as total biomass of queen-produced sexuals (new queens and males), was significantly positively associated with queen longevity, measured from the day the first worker was produced. We suggest that a positive relationship between reproductive success and longevity was inherited from nonsocial ancestors showing parental care and maintained, at least in part, because the presence of workers buffers queens against extrinsic mortality. PMID- 19298497 TI - Plasma folate and total homocysteine levels are associated with the risk of myocardial infarction, independently of each other and of renal function. AB - OBJECTIVES: To investigate the relationship between plasma folate, vitamin B12 and total homocysteine concentrations, dietary intake of folate and vitamins B12, B6 and B2, and the risk of first acute myocardial infarction (MI). DESIGN: Nested case-referent study with up to 13 years of follow-up. SETTING: The population based Northern Sweden Health and Disease Study, with 73 879 participants at the time of case ascertainment. SUBJECTS: A total of 571 MI cases (406 men) and 1569 matched referents. Of the cases, 530 had plasma samples available, and 247 had dietary B-vitamin intake data. RESULTS: Plasma concentrations of folate were inversely associated, and total homocysteine positively associated, with the risk of MI, independently of each other and of a number of established and novel cardiovascular risk factors, including renal function [multivariate odds ratio for highest vs. lowest quintile of folate 0.52 (95% CI 0.31-0.84), P for trend = 0.036, and homocysteine 1.92 (95% CI 1.20-3.09), P for trend = 0.006]. For plasma vitamin B12 concentrations, and vitamin B12, B6 and B2 intake, no clear risk relationship was apparent. Though not statistically significant, the results for folate intake were consistent with those for plasma concentrations. CONCLUSIONS: In this large prospective study of a population without mandatory folic acid fortification, both folate and homocysteine were strongly associated with the risk of myocardial infarction, independently of each other and of renal function. Although randomized trials of folic acid supplementation are needed to determine causality, our findings highlight the potential importance of folate, or sources of folate, in incident cardiovascular disease. PMID- 19298496 TI - Intensive lipid lowering may reduce progression of carotid atherosclerosis within 12 months of treatment: the METEOR study. AB - BACKGROUND: In several statin trials, vascular event rates for treatment groups begin to separate 1 year after commencement of treatment. For atherosclerosis progression, the temporal sequence of the effect has not been defined. We used data from the Measuring Effects on intima media Thickness: an Evaluation Of Rosuvastatin (METEOR) trial to determine the earliest time point at which significant differences in atherosclerosis progression rates could be detected after initiation of statin therapy. METHODS: The METEOR trial was a double-blind, randomized placebo-controlled trial that studied the effect of LDL-C lowering with 40 mg rosuvastatin on the rate of change of carotid intima media thickness (CIMT) measured by B-mode ultrasound amongst 984 low risk subjects. Ultrasound assessments were made at baseline and every 6 months up to 2 years. RESULTS: Rosuvastatin treatment was associated with a 49% reduction in LDL-C-C, a 34% reduction in total cholesterol, an 8.0% increase in HDL-C and a 16% reduction in triglycerides (all P < 0.0001 compared with placebo). The difference in rate of mean maximum CIMT progression between the rosuvastatin and placebo groups (based on near and far wall measurements from both left and right common carotid and internal carotid segments and carotid bifurcation) was not statistically significant after 6 months (0.0023 mm year(-1) and 0.0106 mm year(-1), respectively P = 0.34). After 12 months, CIMT progression rates were significantly different between the groups: 0.0032 mm year(-1) and 0.0133 mm year(-1) in the rosuvastatin-treated and placebo-treated groups, respectively (P = 0.049). This divergence grew with further follow-up: -0.0009 mm year(-1) and 0.0131 mm year(-1) after 18 months (P < 0.001) and -0.0014 mm year(-1) and 0.0131 mm year(-1) after 24 months of treatment (P < 0.001). Results were stronger for the mean common CIMT progression (based on near and far wall measurements from both left and right common carotid segments). CONCLUSION: Aggressive LDL-C lowering seems to exert its beneficial effect on atherosclerosis progression during the first 12 months of treatment. This parallels the timing of event reduction seen in clinical trials and suggests that the efficacy of lipid lowering treatment on CIMT progression can be evaluated in trials with a duration of 1 year, given sufficient sample size, high precision of measurements and a treatment effect comparable to that seen in METEOR. PMID- 19298498 TI - Temporal discrepancies in the association between the apoB/apoA-I ratio and mortality in incident dialysis patients. AB - BACKGROUND: In the general population, a high apoB/apoA-I ratio is a strong risk factor for cardiovascular disease and mortality. However, whether this is the case in chronic kidney disease (CKD) patients is currently unknown. STUDY DESIGN: The apoB/apoA-I ratio was evaluated in 391 incident CKD stage 5 patients examined close to dialysis initiation, and again after 1 year of dialysis in a subgroup of 182 patients, subsequently followed for up to 3 years. RESULTS: Baseline values of the apoB/apoA-I ratio as well as changes in the ratio during the first year of dialysis correlated with body mass index (BMI) and fat mass. The baseline apoB/apoA-I ratio showed no association with 4-year mortality. However, after adjustment for confounders, a high apoB/apoA-I ratio (>0.9) predicted short-term (first year) survival [hazard ratio (HR): 0.35; 95% confidence interval (CI): 0.13-0.85)] and long-term (next 3 years) mortality (HR: 1.72; 95% CI: 1.01-2.96). An increase in the apoB/apoA-I ratio during the first year of dialysis was linked to a survival advantage thereafter (HR: 0.48; 95% CI: 0.22-0.98). However, this association lost its significance (HR: 0.62; 95% CI: 0.26-1.36) after adjustment for indices of protein-energy wasting. CONCLUSIONS: A high apoB/apoA-I ratio and an increase in this ratio during the first year on dialysis were associated with short-term survival advantage in CKD patients. This paradoxical relationship represents an example of the so-called reverse epidemiology phenomenon in CKD patients and suggests that the apoB/apoA-I ratio should always be interpreted with caution in this patient population. PMID- 19298499 TI - Redundant protective barriers: minimizing operator occupational risk. AB - BACKGROUND: The ubiquitous use of less invasive therapeutic angiographic procedures has created the milieu for long-term occupational risk of cancer and genetic defects. This study set out to determine the relative effectiveness of redundant radiation protective barriers and their impact on operator total-body ionizing radiation exposure in the catheterization suite. METHODS: Thermolucent dosimeter x-irradiation was measured inside and outside personal and movable protective barriers used concurrently during 50 consecutive procedures by a single operator. Additionally, the entrance/exit doses were recorded on the back and chest for all patients to provide insight into radiation scatter patterns. RESULTS: The x-ray beam had an average 90.8% decrement in energy traversing the patient's chest when entrance and exit doses were compared, suggesting a 3.3-fold greater operator scatter radiation exposure below the table compared with that above the table. All 0.5-mm lead equivalent personal barriers reduced operator exposure by 72-95%, whereas the 1.0-mm leaded personal barrier (overlapping gown) reduced exposure by 96%. The 0.75-mm leaded glasses reduced exposure to the left eye by 67%. A leaded left-hand glove reduced exposure by only 20%. The effective calculated operator radiation exposure risk reduction provided by the use of personal and movable barriers reduced the theoretical risk of fatal or nonfatal cancer by 22-fold while decreasing potential severe genetic effect by 25-fold in comparison to movable barriers alone. CONCLUSIONS: The optimal use of combined personal and movable (redundant) lead barriers results in a significant reduction in total-body operator radiation exposure in the catheterization laboratory. The use of redundant barriers in the catheterization suite is associated with a dramatic theoretical long-term occupational risk reduction and should be encouraged. PMID- 19298500 TI - Determining the in-hospital cost of bleeding in patients undergoing percutaneous coronary intervention. AB - BACKGROUND: The economic impact of bleeding in the setting of nonemergent percutaneous coronary intervention (PCI) is poorly understood and complicated by the variety of bleeding definitions currently employed. This retrospective analysis examines and contrasts the in-hospital cost of bleeding associated with this procedure using six bleeding definitions employed in recent clinical trials. METHODS: All nonemergent PCI cases at Christiana Care Health System not requiring a subsequent coronary artery bypass were identified between January 2003 and March 2006. Bleeding events were identified by chart review, registry, laboratory, and administrative data. A microcosting strategy was applied utilizing hospital charges converted to costs using departmental level direct cost-to-charge ratios. The independent contributions of bleeding, both major and minor, to cost were determined by multiple regression. Bootstrap methods were employed to obtain estimates of regression parameters and their standard errors. RESULTS: A total of 6,008 cases were evaluated. By GUSTO definitions there were 65 (1.1%) severe, 52 (0.9%) moderate, and 321 (5.3%) mild bleeding episodes with estimated bleeding costs of $14,006; $6,980; and $4,037, respectively. When applying TIMI definitions there were 91 (1.5%) major and 178 (3.0%) minor bleeding episodes with estimated costs of $8,794 and $4,310, respectively. In general, the four additional trial-specific definitions identified more bleeding events, provided lower estimates of major bleeding cost, and similar estimates of minor bleeding costs. CONCLUSIONS: Bleeding is associated with considerable cost over and above interventional procedures; however, the choice of bleeding definition impacts significantly on both the incidence and economic consequences of these events. PMID- 19298501 TI - Collagen mechanics: a rationale for radiofrequency energy to treat mitral regurgitaton. AB - Despite a variety of pharmacological interventions to limit the regurgitant fraction, mitral regurgitation requires surgical intervention for definitive treatment. New nonsurgical techniques are being applied to treat mitral regurgitation. The mitral and tricuspid annuli are areas of dense collagen. Application of heat, in the form of radiofrequency energy, can be used to shrink mitral valve annuli and offers a promising approach to the treatment of mitral regurgitation. PMID- 19298502 TI - Support for family carers of children and young people with developmental disabilities and challenging behaviour: what stops it being helpful? AB - BACKGROUND: Many family carers find the support they receive in respect of their child's challenging behaviour unhelpful. This study sought to identify carer perceptions of the ways in which support is unhelpful and how it could be more helpful. METHODS: Thirteen mothers, caring for a child with intellectual disability and challenging behaviour, were interviewed. Parental perceptions and concerns regarding support received were investigated. Transcribed interviews were analysed using interpretive phenomenological analysis. RESULTS: Parents reported problems with generic disability services including accessing good services, obtaining relevant information, working relationships with professionals and issues with respite provision. Concerns were also expressed about challenging behaviour-specific provision including ineffective strategies being suggested, an apparent lack of expertise, insufficient input and their child's exclusion from services. CONCLUSIONS: More preventative approaches, more widespread adoption of effective behaviour management and improved partnership between professionals and families appear needed. Increasing family support may be ineffective if not accompanied by greater insight into the factors related to effectiveness and recognition of the role of informal support. PMID- 19298503 TI - Staff variables associated with the challenging behaviour of clients with severe or profound intellectual disabilities. AB - BACKGROUND: Previous research has identified that staff-client interactions play an important role in the origin and maintenance of challenging behaviour. Particularly, the reciprocity between staff behaviour and client behaviour has been considered a key issue. Furthermore, severe challenging behaviour has been found to elicit negative emotional reactions from staff which in turn may influence staff's behaviour. Another variable that has been associated with staff behaviour are staff's attributions regarding clients' challenging behaviour. The present study tested several hypotheses about associations between staff variables and challenging behaviour. METHOD: Questionnaires were used to investigate associations between the attributions, emotional reactions and behavioural reactions of 51 staff members towards challenging behaviour of clients with severe or profound intellectual disabilities who displayed self injurious behaviour, stereotyped behaviour and/or aggressive/destructive behaviour. RESULTS: Staff members reported that reactions to challenging behaviour differed according to the type of challenging behaviour. Negative emotional reactions were positively associated with challenging behaviour. Associations between emotional reactions, staff beliefs and staff reactions were inconsistent. CONCLUSIONS: The findings suggest that there is a need to look for a better conceptualization and assessment of the variables under investigation. PMID- 19298504 TI - Clonal nature of odontogenic tumours. AB - BACKGROUND: Although clonal origin is an essential step in the comprehension of neoplasias, there have been no studies to examine whether odontogenic tumours are derived from a single somatic progenitor cell. The purpose of this study was to investigate the clonal origin of odontogenic tumours. METHODS: Fresh samples of seven ameloblastomas, two odontogenic mixomas, two adenomatoid odontogenic tumour, one calcifying odontogenic cyst, one calcifying epithelial odontogenic tumour (CEOT) and six odontogenic keratocyst (OKC) of female patients were included in this study. After DNA extraction, the HUMARA gene polymorphism assay was performed. RESULTS: Most of the informative odontogenic lesions studied (12 out of 16) showed a monoclonal pattern. Among the polyclonal cases, two were OKC, one CEOT and one odontogenic mixoma. CONCLUSIONS: Our results suggest that most odontogenic tumours are monoclonal. PMID- 19298505 TI - Oral health and related quality of life status in patients from UK and Turkey: a comparative study in Behcet's disease. AB - BACKGROUND: The aim of this study was to evaluate and compare oral health-related quality of life (oral QoL) in patients from UK and Turkey with Behcet's disease (BD). METHODS: Thirty-one BD patients from UK (F/M: 18/13, mean age: 41.8 +/- 11.5 years) and Turkey (F/M: 18/13, mean age: 41.5 +/- 10.3) who were matched according to age and gender were included in the study. All patients had active oral ulcers. Oral QoL was assessed by Oral Health Impact Profile-14 (OHIP-14). Oral health was evaluated by dental and periodontal indices. RESULTS: No significant difference was found in OHIP-14 scores between patients from UK (22.7 +/- 14.4) and Turkey (20.4 +/- 14.3) (P = 0.709). The OHIP-14 score correlated with the healing time of oral ulcers in UK (r = 0.4, P = 0.04) and the number of oral ulcers in Turkey (r = 0.4, P = 0.012). The number of oral ulcers per month was significantly higher in UK (3.3 +/- 2.8) compared with that in Turkey (1.5 +/ 2.5) (P = 0.014). However, the number of filled teeth and frequency of tooth brushing were significantly lower in patients from Turkey compared with those in UK (P = 0.000). Similarly, the duration since the last dental visit (5.1 +/- 7.2 months) was significantly lower in UK compared with that in Turkey (28.6 +/- 23.7 months) (P = 0.000). CONCLUSIONS: Oral QoL was similar in patients from UK and Turkey with active oral ulcers. However, the number of oral ulcers was observed to be higher in UK. As expected, a lower utilization rate of dental services might have led to a poorer oral health in patients from Turkey. PMID- 19298506 TI - Microbial community analysis at crude oil-contaminated soils targeting the 16S ribosomal RNA, xylM, C23O, and bcr genes. AB - AIMS: The analyses targeting multiple functional genes were performed on the samples of crude oil-contaminated soil, to investigate community structures of organisms involved in monoaromatic hydrocarbon degradation. METHODS AND RESULTS: Environmental samples were obtained from two sites that were contaminated with different components of crude oil. The analysis on 16S rRNA gene revealed that bacterial community structures were clearly different between the two sites. The cloning analyses were performed by using primers specific for the catabolic genes involved in the aerobic or anaerobic degradation of monoaromatic hydrocarbons, i.e. xylene monooxygenase (xylM), catechol 2,3-dioxygenase (C23O), and benzoyl CoA reductase (bcr) genes. From the result of xylM gene, it was suggested that there are lineages specific to the respective sites, reflecting the differences of sampling sites. In the analysis of the C23O gene, the results obtained with two primer sets were distinct from each other. A comparison of these suggested that catabolic types of major bacteria carrying this gene were different between the two sites. As for the bcr gene, no amplicon was obtained from one sample. Phylogenetic analysis revealed that the sequences obtained from the other sample were distinct from the known sequences. CONCLUSIONS: The differences between the two sites were demonstrated in the analyses of all tested genes. As for aerobic cleavage of the aromatic ring, it was also suggested that analysis using two primer sets provide more detailed information about microbial communities in the contaminated site. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study demonstrated that analysis targeting multiple functional genes as molecular markers is practical to examine microbial community in crude oil-contaminated environments. PMID- 19298507 TI - Heat resistance and the effects of continuous pasteurization on the inactivation of Byssochlamys fulva ascospores in clarified apple juice. AB - AIMS: To determine thermal resistance, the effect of pasteurization temperature variations (c. 2 degrees C) in a continuous system in the number of decimal reductions (n) of a Byssochlamys strain in clarified apple juice (CAJ). METHODS AND RESULTS: Thermal destruction kinetics of Byssochlamys fulva IOC 4518 in thermal death tubes were determined at 85 degrees , 90 degrees , 92 degrees and 95 degrees C by using Weibull distribution frequency model. Three processes with different heating and holding temperatures (A: 94 degrees , 92 degrees C; B: 95 degrees , 93 degrees C; C: 96 degrees , 94 degrees C, respectively) were performed in a continuous system. Process time was 30 s. delta (time of first decimal reduction) values were: 42.98, 8.10, 3.62 and 1.81 min. Variable n ranged from 0.16 to >4.78 for process B (equivalent to industrial). Variable n (0.95 2.66 log CFU ml(-1)) were obtained in CAJ bottles processed under condition B, while process A resulted in total heat-resistant mould (HRM) survival and process C in total HRM destruction. CONCLUSIONS: This study demonstrates that small variations in temperature during the CAJ pasteurization could result in elimination or survival of HRM due to its nonlogarithmic behaviour. SIGNIFICANCE AND IMPACT OF THE STUDY: This was the first study to use Weibull frequency method to model inactivation of HRM in fruit juices. Temperature variations could culminate in the presence of HRM in pasteurized juices even when low counts (<10 spores per 100 ml) were present in the raw materials. PMID- 19298508 TI - Survival of Erwinia amylovora in mature apple fruit calyces through the viable but nonculturable (VBNC) state. AB - AIMS: Survival of Erwinia amylovora, causal agent of fire blight in pome fruits and other rosaceous plants, was monitored inside mature apples calyces under some storage conditions utilized in fruit. METHODS AND RESULTS: Apple fruit calyces inoculated with two E. amylovora strains and their respective GFP-marked strains were maintained at 26 degrees and 5 degrees C, and the effect of copper treatment was assayed at 0.01 and 0.1 mmol l(-1) CuSO4. In nontreated apples at 26 degrees C, part of the population of E. amylovora survived in the 'viable but nonculturable' (VBNC) state, whereas at 5 degrees C the majority of the population retained culturability. In copper-treated apples, the whole population adopted the VBNC state irrespective of temperature. Regardless of temperature, copper and inoculum dose, VBNC cells recovered culturability and pathogenicity in King's B broth or by host plant passage. CONCLUSIONS: Erwinia amylovora survived for at least 35 days in mature apple calyces. Besides, the ability of the pathogen in the VBNC state to regain culturability and pathogenicity suggests that the apple fruit could be a potential carrier of E. amylovora contributing to the spreading of fire blight disease. SIGNIFICANCE AND IMPACT OF THE STUDY: The risk of E. amylovora dissemination through mature fruit transport, although low, has been demonstrated, and should be considered in pest risk assessments. PMID- 19298509 TI - Co-cultivation of a bacteriocin-producing mixed culture of Bifidobacterium thermophilum RBL67 and Pediococcus acidilactici UVA1 isolated from baby faeces. AB - AIMS: We investigated the kinetics of growth, metabolism and antimicrobial activity of Bifidobacterium thermophilum RBL67 and Pediococcus acidilactici UVA1 previously isolated as a consortium from human baby faeces and producing antimicrobial proteinaceous compounds. METHODS AND RESULTS: Cell growth, antimicrobial activity, glucose consumption, organic acid production and pediocin gene expression were monitored during pure and mixed strain batch cultures with controlled pH (6.0) at 37 degrees C. The balance of the two strains in mixed cultures was stable, yielding high cell count of 10(9) CFU ml(-1) after a very short incubation time of 4 h for UVA1 and 7 h for RBL67, and RBL67 was not affected by the high production of pediocin by UVA1 during co-culture (up to 12.8 microg ml(-1)). Furthermore, a real-time PCR assay was developed and allowed gene expression analysis of the pediocin-gene pedA. CONCLUSIONS: The co-culture of RBL67 and UVA1 showed high stability, cell yields and bacteriocin production during batch cultures and has potential for use as a probiotic mixture with antibacterial properties. In addition, a gene-expression real-time PCR assay was successfully developed, used for the relative quantification of the pediocin transcript pedA and demonstrated to be a valuable complementary tool to activity assay. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study reporting of a stable mixed culture of two bacteriocin-producing strains of human origin. PMID- 19298510 TI - Characterization of the bacterial spoilage flora in marinated pork products. AB - AIMS: To investigate the microbiota in marinated, vacuum-packed pork and to characterize isolated bacteria with regard to their spoilage potential. METHODS AND RESULTS: Laboratory marinated pork meat and commercial products from three Norwegian producers were examined. Lactic acid bacteria dominated in all products at the expiration date. The flora in marinated products was similar only for products from the same plant. Strains of Lactobacillus algidus, Lactobacillus sakei, Lactobacillus curvatus, Carnobacterium divergens, Carnobacterium maltaromaticum, Leuconostoc mesenteroides, Leuconostoc carnosum and Leuconostoc sp. were isolated and tested for their spoilage potential. Samples inoculated with Lact. algidus or Leuc. mesenteroides were rated as most unpleasant by randomly selected people. A sensory panel scored samples with Lact. algidus highest for sour and intense odour. Lactobacillus algidus was found in products from two out of three production plants. Culture-independent DNA isolation confirmed that cultivation on Blood agar at 20 degrees C yielded a representative picture of the total flora in marinated flintsteak. CONCLUSIONS: Lactobacillus algidus may be an important, but underestimated, spoilage organism that needs to be focused on more when spoilage of vacuum-packed meat is considered. SIGNIFICANCE AND IMPACT OF THE STUDY: Routine microbial testing may have to be revised in order to detect spoilage LAB that are unable to grow under currently used conditions. PMID- 19298511 TI - Evaluation of murine norovirus as a surrogate for human norovirus and hepatitis A virus in heat inactivation studies. AB - AIMS: To determine the suitability of murine norovirus (MNV) as a surrogate for human norovirus (HuNoV) in heat inactivation studies. METHODS AND RESULTS: MNV, hepatitis A virus (HAV) and HuNoV genogroup I and II (GI and GII) specific real time quantitative reverse transcription (qRT)-PCR assays were used to determine the effects of heat exposure (63 and 72 degrees C) for up to 10 min in water and milk. Using culture assays, MNV and HAV showed similar reductions in infectivity over time. Both HuNoV GI and GII showed lower log reductions in qRT-PCR titre following heat exposure than either MNV or HAV. No significant protective effect of milk was observed for any virus. CONCLUSIONS: MNV is as suitable a surrogate for HuNoV as HAV. In heat inactivation studies at 63 and 72 degrees C, qRT-PCR results indicate that HuNoV is less susceptible to heat than either HAV or MNV and so neither virus may be an appropriate surrogate for HuNoV. SIGNIFICANCE AND IMPACT OF THE STUDY: Caution should be used when extrapolating surrogate virus data for HuNoV. Although not conclusive, our results suggest that HuNoV may be more resistant to heat than either HAV or MNV. PMID- 19298512 TI - Heterogeneous shedding of Brucella abortus in milk and its effect on the control of animal brucellosis. AB - AIMS: To ascertain whether in Brucella abortus-infected water buffalo herds, the number of newly infected animals could be reduced by culling superspreaders (the animals secreting > or =10(4) CFU per ml of milk). METHODS AND RESULTS: The number of B. abortus present in the milk (CFU per ml) from 500 water buffaloes was measured by the culture. Each animal was tested three times, at one month intervals. The presence or the absence of B. abortus in each milk sample was confirmed by PCR. A majority of infected animals shed the pathogen at a low level (< or =10(3) CFU ml(-1)). However, a few infected individuals (superspreaders) shed large numbers of B. abortus (> or =10(4) CFU ml(-1)). Quantitative PCR of B. abortus positive milk samples gave comparable results to culture. Culling of the superspreaders was sufficient to arrest the spread of infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The approach described here can reduce significantly the cost of controlling brucellosis. Culture and quantitative PCR tests identify superspreaders and, compared with the serological tests in use to detect brucellosis, provide also a more accurate estimate of the disease incidence. PMID- 19298513 TI - Antimicrobial activity of Coniothyrium minitans and its macrolide antibiotic macrosphelide A. AB - AIMS: Assessment of antimicrobial activity of the mycoparasite Coniothyrium minitans and its macrolide antibiotic macrosphelide A. METHODS AND RESULTS: Thirteen isolates of C. minitans were tested for ability to inhibit a number of filamentous fungi, yeasts, oomycetes and bacteria in agar based tests. Activity was found against some ascomycetes, basidiomycetes, oomycetes and Gram-positive bacteria, but not against zygomycetes, yeasts or Gram-negative bacteria tested. Six C. minitans isolates (Conio, Contans, IVT1, CM/AP/3118, B279/1, A1/327/1) were found to produce macrosphelide A in liquid culture and no other antibiotics were detected. On agar, macrosphelide A inhibited growth of some ascomycetes, basidiomycetes, oomycetes and all four Gram-positive bacteria tested, including the medically important Staphylococcus aureus with a minimum inhibitory concentration of < or =500 microg ml(-1). There was no inhibition observed against the yeasts and Gram-negative bacteria when macrosphelide A was tested at 700 microg ml(-1). CONCLUSIONS: The spectrum and level of activity of macrosphelide A produced by C. minitans against micro-organisms are extended markedly compared to previous reports. SIGNIFICANCE AND IMPACT OF THE STUDY: Macrosphelide A was effective against Staph. aureus. Further study on the control of this bacterium is merited in view of the development of antibiotic resistance. PMID- 19298514 TI - Characterization of a methane-utilizing strain and its application for monitoring methane. AB - AIMS: To explore new resources of methane-utilizing micro-organism and develop a microbial biosensing system for monitoring methane released from natural and semi natural ecosystems. METHODS AND RESULTS: A methane (CH(4))-utilizing bacterial strain was isolated from paddy soil using CH(4) as the sole carbon source and identified as Klebsiella sp. ME17 by phenotyping and 16S rDNA sequence analysis. The efficiency of CH(4) utilization of strain ME17 was 83.2% by gas chromatography analysis. A microbial biosensing system for CH(4) detection was developed by combining immobilized cells of strain ME17 with a dissolved oxygen sensor. It was found that response time of the system to CH(4) was <90s. The dissolved O(2) consumption increased with increasing CH(4) from 0% to 16.0% (v/v) demonstrating a positive linear relationship with a low detection limit of 0.2% (v/v). The relative standard deviation is 3.48%. CONCLUSIONS: Klebsiella sp. ME17 isolate is capable of utilizing CH(4). The microbial biosensing system of strain ME17 has been successfully applied to measure standard CH(4) sample with satisfactory results. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that certain strains of Klebsiella genus are capable of utilizing CH(4). Our proposed method appears very attractive for CH(4) measurement in coal mine. PMID- 19298515 TI - Interactions of plant-beneficial bacteria with the ascomycete Coniochaeta ligniaria. AB - AIMS: To assess the interactions between Coniochaeta ligniaria F/TGF15 obtained from torrefied grass fibers (TGF) and selected bacteria from the same substrate. METHODS AND RESULTS: Upon coinoculation on potato dextrose agar, Pseudomonas putida 15/TGE5, Methylobacterium radiotolerans 56/TGF10, Serratia plymutica 23/TGE5, Pseudomonas corrugata 31/TGE5, Leifsonia xyli subsp. xyli 66/TGF10, Mycobacterium anthracenicum 70/TGF15 and Agromyces aurantiacus 95/TGF15 were translocated by C. ligniaria, but not in the absence of the fungus. Pseudomonas putida, P. corrugata, L. xyli subsp. xyli and A. aurantiacus were able to grow on compounds released by the fungus, but not M. radiotolerans. Antagonism towards C. ligniaria was observed for S. plymutica and P. corrugata. Pseudomonas putida was translocated by the fungus in TGF up to at least 45 mm. It also multiplied on the hyphae of C. ligniaria in TGF, reaching CFU densities of log 8.4 g(-1) dry TGF in 20 d, while the strain could not grow in nonfungal TGF. Methylobacterium radiotolerans was not translocated by the fungus in TGF. CONCLUSIONS: Several of the selected bacteria could grow on the compounds released by the fungus, whereas two bacteria inhibited or killed the fungus. SIGNIFICANCE AND IMPACT OF THE STUDY: It is shown that C. ligniaria has a dual role in bacterial colonization of TGF, being crucial for the detoxification of TGF, meanwhile stimulating growth and translocation of a consortium of plant-growth-promoting bacteria. PMID- 19298516 TI - Effects of reduction in beef surface water activity on the survival of Salmonella Typhimurium DT104 during heating. AB - AIM: To investigate the influence of reducing beef surface water activity (a(w)) on the survival of Salmonella Typhimurium DT104 during heating. METHODS AND RESULTS: Beef discs were surface inoculated with S. Typhimurium DT104 and either untreated or dried to achieve surface a(w) values of 0.95, 0.85 and 0.70. The samples were vacuum packed, heat-treated at 60 degrees C and removed at predetermined times. The inactivation curves were influenced by a(w) and treatment time. Biphasic inactivation curves were observed for S. Typhimurium DT104 heat-treated on beef samples with altered a(w) values, which were characterized by an initial decline in cell numbers at commencement of heating followed by a much slower rate of inactivation during the remaining treatment period. Point estimates of the heating time required to achieve a 1 log reduction on beef surfaces with a(w) of 0.99, 0.95, 0.85 and 0.70 were 0.5, 1.55, 11.25 and 17.79 min, respectively. CONCLUSIONS: A decrease in beef surface a(w) can substantially enhance the survival of S. Typhimurium DT104 after heating. SIGNIFICANCE AND IMPACT OF THE STUDY: Caution needs to be taken using dry air as a decontamination method as this may rapidly decrease product surface and pathogen a(w) values resulting in enhanced survival. PMID- 19298517 TI - Assessment of the stability of human viruses and coliphage in groundwater by PCR and infectivity methods. AB - AIM: To investigate the potential health hazard from infectious viruses where coliphages, or viruses by polymerase chain reaction (PCR), have been detected in groundwater. Two aspects were investigated: the relationship between infectivity and detection by PCR and the stability of coliphage compared to human viruses. METHODS AND RESULTS: Virus decay (1 year) and detection (2 years) studies were undertaken on groundwater at 12 degrees C. The order of virus stability from most to least stable in groundwater, based on first-order inactivation, was: coliphage PhiX174 (0.5 d(-1)) > adenovirus 2 > coliphage PRD1 > poliovirus 3 > coxsackie virus B1 (0.13 d(-1)). The order for PCR results was: norovirus genotype II > adenovirus > norovirus genotype I > enterovirus. CONCLUSIONS: Enterovirus and adenovirus detection by PCR and the duration of infectivity in groundwater followed similar trends over the time period studied. Adenovirus might be a better method for assessing groundwater contamination than using enterovirus; norovirus detection would provide information on a significant human health hazard. Bacteriophage is a good alternative indicator. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR is a useful tool for identifying the health hazard from faecal contamination in groundwater where conditions are conducive to the survival of viruses and their nucleic acid. PMID- 19298518 TI - Concurrent selection for microbial suppression of Fusarium graminearum, Fusarium head blight and deoxynivalenol in wheat. AB - AIM: Identify biological agents that can both control Fusarium head blight (FHB) and reduce deoxynivalenol (DON) production. METHODS AND RESULTS: Concurrent screening methods were used to progressively select soil and food micro-organisms for the ability to suppress Fusarium graminearum, FHB and DON production. The micro-organisms were assessed using up to five assays including: a co-culture and dual-culture assay, an indirect impedance assay, a wheat floret assay, and two assays assessing DON production. Paenibacillus polymyxa W1-14-3 and C1-8-b gave the greatest inhibition of F. graminearum and reduction of DON production in greenhouse evaluations. Compared to a control treatment, they reduced disease severity by 56.5 and 55.4%, F. graminearum colonization of wheat heads by 58.8 and 62.4%, DON production by 84.8 and 89.4%, and increased 100-kernel weights by 56.6 and 66.9%, respectively. CONCLUSIONS: The concurrent selection has resulted in promising antagonists that may possess multiple modes of action, and have the ability to colonize wheat heads in controlled environments. SIGNIFICANCE AND IMPACT OF THE STUDY: A novel concurrent screening method was developed for selection of biocontrol agents for FHB. Two isolates of P. polymyxa were selected and identified. Their potential use as biocontrol agents for FHB is highlighted in this study. PMID- 19298519 TI - The anti-angiogenic factor PEDF is present in the human heart and is regulated by anoxia in cardiac myocytes and fibroblasts. AB - Cardiac diseases such as myocardial infarction and heart failure are among the leading causes of death in western societies. Therapeutic angiogenesis has been suggested as a concept to combat these diseases. The biology of angiogenic factors expressed in the heart such as vascular endothelial growth factor (VEGF) is well studied, whereas data on anti-angiogenic mediators in the heart are scarce. Here we study the expression of the anti-angiogenic factor pigment epithelium-derived factor (PEDF) in the human heart and in human cardiac cells. PEDF expression could be detected in human cardiac tissue on the protein and mRNA levels. PEDF mRNA levels were significantly lower in explanted human ischemic hearts as compared to healthy hearts. Our in vitro experiments showed that human adult cardiac myocytes and fibroblasts constitutively secrete PEDF. In addition to anoxic conditions, cobalt chloride, 2,2'dipyridyl and dimethoxally glycine, which stabilize hypoxia inducible factor-alpha decreased PEDF expression. Furthermore we show that PEDF inhibits VEGF-induced sprouting. We have identified PEDF in healthy and ischemic human hearts and we show that PEDF expression is down-regulated by low oxygen levels. Therefore, we suggest a role for PEDF in the regulation of angiogenesis in the heart and propose PEDF as a possible therapeutic target in heart disease. PMID- 19298520 TI - p27(Kip1) as a prognostic factor in breast cancer: a systematic review and meta analysis. AB - The aim of this study was to comprehensively evaluate via a meta-analysis the association between p27 expression and clinical outcome in breast cancer patients. We conducted a meta-analysis of 20 studies (n= 6463 patients) that evaluated the correlation between p27 expression and indicators of breast cancer clinical outcome, including overall survival (OS), disease-free survival (DFS) and relapse-free survival (RFS). Data pooling was performed by RevMan 4.2. A total of 60% (9 of 15) of the studies showed a significant association between p27 high expression and OS, whereas 25% (2 of 8) and 60% (3 of 5) studies demonstrated a correlation between p27 high expression and DFS and RFS, respectively. The relative risks (RRs) were 1.34 (1.26-1.42) for OS (P < 0.00001), 1.27 (1.10-1.47) for DFS (P= 0.001) and 1.49 (0.92-2.42) for RFS (P= 0.10). In lymph node-negative breast cancer patients, the RRs for OS and RFS were 1.84 (1.30-2.59; P= 0.0005) and 1.30 (0.20-8.50; P= 0.78), respectively. In lymph node-positive breast cancer patients, the RRs for OS and RFS were 2.99 (1.77 5.07; P < 0.0001) and 1.49 (0.80-2.77; P= 0.21), respectively. This meta-analysis indicates that reduced p27 is an independent prognostic factor for poor overall and disease-free cancer survival. PMID- 19298521 TI - Intracrine androgenic apparatus in human bone marrow stromal cells. AB - It was suggested that human mesenchymal stromal cells might contain an intracrine enzyme machinery potentially able to synthesize the cell's own supply of dihydrotestosterone (DHT) from dehydroepiandrosterone (DHEA) pro-hormone produced in the adrenal cortex in the reticular zone, which is unique to primates. Indeed, 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 5alpha-reductase enzyme proteins were expressed in resting mesenchymal stromal cells (MSCs) in vitro. However, the 'bridging' enzymes 17beta-HSDs, catalysing interconversion between 17beta-ketosteroids and 17beta-hydroxysteroids, were not found in resting MSCs, but 17beta-HSD enzyme protein was induced in a dose-dependent manner by DHEA. Quantitative real-time polymerase chain reactions disclosed that this was mainly due to induction of the isoform 5 catalysing this reaction in 'forward', androgen bound direction (P < 0.01). This work demonstrates that the MSCs have an intracrine machinery to convert DHEA to DHT if and when challenged by DHEA. DHEA as substrate exerts a positive, feed-forward up-regulation on the 17beta-hydroxy steroid dehydrogenase-5, which may imply that DHEA-DHT tailor-making in MSCs is subjected to chronobiological regulation. PMID- 19298522 TI - Modulation of cardiac ionic homeostasis by 3-iodothyronamine. AB - 3-iodothyronamine (T(1)AM) is a novel endogenous relative of thyroid hormone, able to interact with trace amine-associated receptors, a class of plasma membrane G protein-coupled receptors, and to produce a negative inotropic and chronotropic effect. In the isolated rat heart 20-25 microM T(1)AM decreased cardiac contractility, but oxygen consumption and glucose uptake were either unchanged or disproportionately high when compared to mechanical work. In adult rat cardiomyocytes acute exposure to 20 microM T(1)AM decreased the amplitude and duration of the calcium transient. In patch clamped cardiomyocytes sarcolemmal calcium current density was unchanged while current facilitation by membrane depolarization was abolished consistent with reduced sarcoplasmic reticulum (SR) calcium release. In addition, T(1)AM decreased transient outward current (I(to)) and I(K1) background current. SR studies involving 20 microM T(1)AM revealed a significant decrease in ryanodine binding due to reduced B(max), no significant change in the rate constant of calcium-induced calcium release, a significant increase in calcium leak measured under conditions promoting channel closure, and no effect on oxalate-supported calcium uptake. Based on these observations we conclude T(1)AM affects calcium and potassium homeostasis and suggest its negative inotropic action is due to a diminished pool of SR calcium as a result of increased diastolic leak through the ryanodine receptor, while increased action potential duration is accounted for by inhibition of I(to) and I(K1) currents. PMID- 19298523 TI - Healthy human salivary glands contain a DHEA-sulphate processing intracrine machinery, which is deranged in primary Sjogren's syndrome. AB - Sjogren's syndrome (SS) patients have low salivary dehydroepiandrosterone (DHEA) and androgen biomarker levels, but high salivary oestrogen levels. The hypothesis was that the healthy glands contain DHEA-sulphate processing intracrine machinery; the local androgen/oestrogen imbalance suggests that this is disarranged in SS. Indirect immunofluorescence and quantitative real-time PCR (qRT-PCR) of steroid sulphatase, sulfotransferase, 3beta- and 17beta hydroxysteroid dehydrogenases (3beta- and 17beta-HSD), 5alpha-reductase and aromatase were performed for labial salivary glands of healthy controls and persons with SS. In control acini steroid sulphatase and sulfotransferase immunoreactivities were located in the basolateral cell parts. 3Beta- and 17beta HSD formed strong, interrupted bands along the basal cell parts. 5alpha-reductase was mainly located in acinar cell nuclei and aromatase in the apical cell membrane. All enzymes were more widespread in ducts. In SS, steroid sulphatase was weak and deranged, 3beta- and 17beta-HSD had lost their strict basal acinar cell localization and 5alpha-reductase was mainly found in the cytoplasm of the acinar cells, whereas aromatase showed similar staining in SS and controls. qRT PCR of labial salivary glands disclosed all corresponding enzyme mRNAs with the levels of 3beta-HSD in SS being the lowest. Healthy tubuloacinar epithelial cells contain complete intracrine machineries for DHEA(-sulphate) pro-hormone processing. These enzymes have in healthy acini an organized architecture, which corresponds with DHEA uptake from the circulation, nuclear site of production of the active dihydrotestosterone (DHT) end product and production of oestrogens into saliva for export to ductal and oral epithelial cells. SS is characterized by low 3beta-HSD levels, which together with impaired subcellular compartmentalization of HSDs and 5alpha-reductase may explain the low local DHT and androgen biomarker levels in SS. PMID- 19298524 TI - Functional characterization and pharmacological rescue of melanocortin-4 receptor mutations identified from obese patients. AB - As the most common monogenic form of human obesity, about 130 naturally occurring melanocortin-4 receptor (MC4R) gene mutations have been identified. In this study, we reported detailed functional characterization of 10 novel human MC4R (hMC4R) mutants including R7C, C84R, S127L, S136F, W174C, A219V, P230L, F261S, I317V and L325F. Flow cytometry experiments showed that six mutants, including R7C, C84R, S127L, W174C, P230L and F261S, have decreased cell surface expression. The other four mutants are expressed at similar levels as the wild-type hMC4R. Binding assays showed that the mutants have similar binding affinities for the agonist and endogenous antagonist agouti-related protein. Signalling assays showed that S136F is defective in signalling. Multiple mutagenesis showed that S136 of hMC4R is required for the normal function of the receptor. To identify potential therapeutic approaches for patients with intracellularly retained MC4R mutants, we tested the effect of an MC4R inverse agonist, ML00253764, on C84R and W174C. We showed that ML00253764 could function as a pharmacological chaperone rescuing the mutant MC4Rs to the cell surface. The rescued mutants are functional with increased cAMP production in response to agonist stimulation. In conclusion, of 10 mutants we studied, 6 had decreased cell surface expression. Pharmacological chaperone is a potential approach for treating obesity caused by MC4R mutations that result in intracellular retention. PMID- 19298525 TI - Development of c-kit immunopositive interstitial cells of Cajal in the human stomach. AB - Interstitial cells of Cajal (ICC) include several types of specialized cells within the musculature of the gastrointestinal tract (GIT). Some types of ICC act as pacemakers in the GIT musculature, whereas others are implicated in the modulation of enteric neurotransmission. Kit immunohistochemistry reliably identifies the location of these cells and provides information on changes in ICC distribution and density. Human stomach specimens were obtained from 7 embryos and 28 foetuses without gastrointestinal disorders. The specimens were 7-27 weeks of gestational age, and both sexes are represented in the sample. The specimens were exposed to anti-c-kit antibodies to investigate ICC differentiation. Enteric plexuses were immunohistochemically examined by using anti-neuron specific enolase and the differentiation of smooth muscle cells (SMC) was studied with anti-alpha smooth muscle actin and anti-desmin antibodies. By week 7, c-kit immunopositive precursors formed a layer in the outer stomach wall around myenteric plexus elements. Between 9 and 11 weeks some of these precursors differentiated into ICC. ICC at the myenteric plexus level differentiated first, followed by those within the muscle layer: between SMC, at the circular and longitudinal layers, and within connective tissue septa enveloping muscle bundles. In the fourth month, all subtypes of c-kit-immunoreactivity ICC which are necessary for the generation of slow waves and their transfer to SMC have been developed. These results may help elucidate the origin of ICC and the aetiology and pathogenesis of stomach motility disorders in neonates and young children that are associated with absence or decreased number of these cells. PMID- 19298526 TI - Endostatin induces autophagy in endothelial cells by modulating Beclin 1 and beta catenin levels. AB - Endostatin is a well-characterized endogenous inhibitor of angiogenesis that affects cell proliferation and migration by inhibiting integrin and Wnt-mediated signalling pathways. Here, we show that endothelial cells treated with native and P125A-endostatin activate autophagy. Because autophagy can either be protective or induce programmed cell death, experiments were carried out to understand the signalling pathways leading to autophagy in endothelial cells. P125A-endostatin treatment increased the levels of Beclin 1, a crucial molecule in vesicle nucleation and autophagy. The treatment also reduced the levels of Bcl-2, Bcl x(L) and beta-catenin; however, progressively increasing amounts of Bcl-2 and Bcl x(L) were found to be complexed with Beclin 1. Increased beta-catenin and Wnt mediated signalling reduced Beclin 1 levels and rescued endothelial cells from endostatin-induced autophagy. Finally, knocking down Beclin 1 levels by RNA interference decreased autophagy and accelerated caspase activation in endostatin treated cells. These studies suggest that endothelial cells may initiate autophagy as a survival response to limit the effects of angiogenesis inhibitors. Thus, interfering with autophagy can potentiate the effects of endostatin by promoting a switch to apoptosis. PMID- 19298527 TI - Survivin is an essential mediator of arthritis interacting with urokinase signalling. AB - Proto-oncogene survivin has recently been identified as a prognostic marker distinguishing patients with destructive rheumatoid arthritis (RA). In the present material of 132 RA patients and 82 controls, the levels of survivin correlated to urokinase (uPA) (r= 0.46), a plasminogen activator over-expressed in inflamed joints and known to exhibit potent arthritogenic properties. Here we evaluate the functional relationship between these proteins using primary synovial fibroblasts and leucocytes of RA patients, human monocytic (THP-1) and fibroblast (MRC-5) cell lines. Using inhibitors of intracellular signalling, we show that uPA and survivin share common transduction pathways in synovial fibroblasts being dependent on the activity of tyrosine kinases, phosphatidylinositide 3 kinase and mitogen effector kinase. Moreover, uPA production is significantly reduced in fibroblasts if survivin synthesis has been silenced by siRNA. Importantly, silencing of survivin in fibroblasts prevented their invasive growth in knee joints of severe combined immune deficient mice. Interaction of uPA with receptor up-regulates survivin expression in leucocytes. In turn, survivin is required for the up-regulation of uPA receptor on the cell surface. These findings indicate that survivin is an essential mediator of arthritogenic properties of uPA regulating its synthesis in synovial fibroblasts and uPAR expression in leucocytes. Close correlation between survivin and uPA levels in patients with RA supports the importance of this connection for the pathogenesis of arthritis. PMID- 19298528 TI - A common hereditary single-nucleotide polymorphism in the gene of FAS and colorectal cancer survival. AB - Apoptosis plays an important role in embryogenesis, autoimmunity and tumourigenesis. Cell surface death receptors such as TNFRSF6 (FAS) confer a major apoptotic effect. A single-nucleotide polymorphism in the FAS promoter gene, 670A/G, modulates apoptotic signalling and has been related to susceptibility and progression of a variety of cancers. The present study aimed to evaluate the role of this polymorphism for survival of patients with colorectal cancer. We performed a retrospective analysis including 433 patients with histologically confirmed colorectal cancer. A Cox regression model including FAS -670 genotypes, age at diagnosis, tumour grading, primary tumour size, number of lymph nodes examined, number of metastatic lymph nodes, tumour stage and application of fluorouracil-based adjuvant chemotherapy was used to estimate the effect of the FAS genotype on survival. FAS -670A/G genotype frequencies were 24.2% (AA), 46.3% (AG) and 29.5% (GG). Forty-nine patients were excluded from the Cox regression analysis because of missing values. Out of the remaining 384 patients, 69 (18%) died during a follow-up of maximum 10 years. Mean follow-up time was 58 +/- 34 months (median 55 months). Carriers of the homozygous FAS -670GG genotype had a significantly lower survival rate compared with AA/AG genotype carriers (relative risk 1.76, 95% confidence interval 1.08-2.87; P= 0.023). The FAS -670A/G polymorphism may be associated with overall survival time of patients with colorectal cancer. PMID- 19298529 TI - Mechanisms underlying fibronectin-induced up-regulation of P2X4R expression in microglia: distinct roles of PI3K-Akt and MEK-ERK signalling pathways. AB - Microglia are resident immune cells in the central nervous system that become activated and produce pro-inflammatory and neurotrophic factors upon activation of various cell-surface receptors. The P2X(4) receptor (P2X(4)R) is a sub-type of the purinergic ion-channel receptors expressed in microglia. P2X(4)R expression is up-regulated under inflammatory or neurodegenerative conditions, and this up regulation is implicated in disease pathology. However, the molecular mechanism underlying up-regulation of P2X(4)R in microglia remains unknown. In the present study, we investigated the intracellular signal transduction pathway that promotes P2X(4)R expression in microglia in response to fibronectin, an extracellular matrix protein that has previously been shown to stimulate P2X(4)R expression. We found that in fibronectin-stimulated microglia, activation of phosphatidylinositol 3-kinase (PI3K)-Akt and mitogen-activated protein kinase kinase (MAPK kinase, MEK)-extracellular signal-regulated kinase (ERK) signalling cascades occurred divergently downstream of Src-family kinases (SFKs). Pharmacological interference of PI3K-Akt signalling inhibited fibronectin-induced P2X(4)R gene expression. Activation of PI3K-Akt signalling resulted in a decrease in the protein level of the transcription factor p53 via mouse double minute 2 (MDM2), an effect that was prevented by MG-132, an inhibitor of the proteasome. In microglia pre-treated with MG-132, fibronectin failed to up-regulate P2X(4)R expression. Conversely, an inhibitor of p53 caused increased expression of P2X(4)R, implying a negative regulatory role of p53. On the other hand, inhibiting MEK-ERK signalling activated by fibronectin suppressed an increase in P2X(4)R protein but interestingly did not affect the level of P2X(4)R mRNA. We also found that fibronectin stimulation resulted in the activation of the translational factor eIF4E via MAPK-interacting protein kinase-1 (MNK1) in an MEK ERK signalling-dependent manner, and an MNK1 inhibitor attenuated the increase in P2X(4)R protein. Together, these results suggest that the PI3K-Akt and MEK-ERK signalling cascades have distinct roles in the up-regulation of P2X(4)R expression in microglia at transcriptional and post-transcriptional levels, respectively. PMID- 19298530 TI - The decrease of expression of ryanodine receptor sub-type 2 is reversed by gentamycin sulphate in vascular myocytes from mdx mice. AB - The mdx mouse, a model of the human Duchenne muscular dystrophy, displays impaired contractile function in skeletal, cardiac and smooth muscles. We explored the possibility that ryanodine receptor (RYR) expression could be altered in vascular muscle. The three RYR sub-types were expressed in portal vein myocytes. As observed through mRNA and protein levels, RYR2 expression was strongly decreased in mdx myocytes, whereas RYR3 and RYR1 expression were unaltered. The use of antisense oligonucleotide directed against RYR sub-types indicated that caffeine-induced Ca(2+) response and Ca(2+) spark frequency depended on RYR2 and RYR1. In mdx mice, caffeine-induced Ca(2+) responses were decreased in both amplitude and maximal rate of rise, and the frequency of Ca(2+) sparks was also strongly decreased. The gentamycin treatment was able to increase both the expression of RYR2 and the caffeine-induced Ca(2+) response to the same level as that observed in wild-type mice. Taken together, these results confirm that both RYR1 and RYR2 are required for vascular Ca(2+) signalling and indicate that inhibition of RYR2 expression may account for the decreased Ca(2+) release from the SR in mdx vascular myocytes. Finally, we suggest that gentamycin can restore the Ca(2+) signalling in smooth muscle from mdx mice by increasing RYR2 and dystrophin expression. These results may help explain the reduced efficacy of contraction in vascular myocytes of mdx mice and Duchenne muscular dystrophy afflicted patients. Gentamycin treatment could be a good therapeutic tool to restore the vascular function. PMID- 19298531 TI - Epigallocatechin-3-gallate attenuates cardiac hypertrophy in hypertensive rats in part by modulation of mitogen-activated protein kinase signals. AB - 1. It has been demonstrated that epigallocatechin-3-gallate (EGCG) inhibits cardiac hypertrophy through its antihypertensive and anti-oxidant effects. However, the underlying molecular mechanism is not clear. 2. In the present study, we tested the hypothesis that EGCG attenuates transaortic abdominal aortic constriction (TAC)-induced ventricular hypertrophy by regulating mitogen activated protein kinase (MAPK) signal pathways in hypertensive rats. Four groups of rats were used: (i) a sham-operated control group; (ii) an EGCG-treated (50 mg/kg per day, i.p., for 21 days) sham-operated group; (iii) a TAC group; and (iv) an EGCG-treated TAC group. Histological analysis of whole hearts and biochemical analyses of left ventricular (LV) tissue were used to investigate the effects of EGCG. 3. The results showed that the LV myocyte diameter and the expression of atrial natriuretic peptide, brain natriuretic peptide and beta myocardial heavy chain were significantly decreased in the EGCG-treated (50 mg/kg per day, i.p.) TAC group. Levels of reactive oxygen species and malondialdehyde in the lV were significantly reduced by EGCG in the TAC group. Total superoxide dismutase, catalase and glutathione peroxidase activities were decreased in the TAC group, and this decrease was significantly restored by EGCG treatment. Phosphorylation of extracellular signal-regulated kinase 2, p38 and c-Jun N terminal kinase 1 was significantly reversed in the LV of EGCG-treated TAC rats (40%, 53% and 52% vs TAC, respectively), accompanied by significant inhibition of nuclear factor-kappaB and activator protein-1. Transaortic abdominal aortic constriction significantly upregulated LV expression of matrix metalloproteinase 9 from 32 +/- 6 to 100 +/- 12% and this increase was inhibited by EGCG treatment (from 100 +/- 12 to 50 +/- 15%). In addition, TAC decreased mitochondrial DNA copy number and the activity of respiratory chain complexes I (from 100 +/- 7 to 68 +/- 5%), III (from 100 +/- 4 to 2 +/- 5%) and IV (from 766 +/- 2 to 100 +/- 5%); this decrease was reversed by EGCG treatment to levels seen in sham-operated rats. PMID- 19298532 TI - Role of activated intrarenal reactive oxygen species and renin-angiotensin system in IgA nephropathy model mice. AB - 1. Using HIGA (high IgA of ddY) mice as an IgA nephropathy model and BALB/c mice as controls, we demonstrated that reactive oxygen species (ROS) and the renin angiotensin system (RAS) were activated in kidneys of HIGA mice. However, it was difficult to establish an association between renal damage and changes in ROS and the RAS. Therefore, the present study was performed to determine whether renal injury is associated with changes in ROS and the RAS in HIGA mice. 2. Male HIGA mice were divided into four groups of 10 each: (i) untreated mice (HIGA + null); (ii) mice treated with the angiotensin AT(1) receptor antagonist olmesartan (5 mg/kg per day; HIGA + OLM); (iii) mice treated with the superoxide dismutase mimetic tempol (50 mg/kg per day; HIGA + Tempol); and (iv) mice treated with RAS independent antihypertensive drugs (30 mg/kg per day hydralazine, 0.6 mg/kg per day reserpine and 12 mg/kg per day hydrochlorothiazide; HIGA + HRH). Mice were treated for 5 weeks. 3. Systolic blood pressure decreased significantly in the HIGA + OLM and HIGA + HRH groups, but not in the HIGA + Tempol group, compared with HIGA + null mice. The expression of two ROS markers (4-hydroxy-2-nonenal and heme oxygenase-1) and angiotensin II as a marker of the RAS decreased significantly in HIGA + OLM and HIGA + Tempol mice, but not in HIGA + HRH mice, compared with HIGA + null mice. As a marker of renal damage, mesangial matrix expansion and the desmin-positive area decreased significantly in the HIGA + OLM and HIGA + Tempol groups, but not in HIGA + HRH group, compared with the HIGA + null group. 4. These data suggest that intrarenal ROS and RAS activation play a pivotal role in the development of IgA nephropathy model mice, from the early phase, independent of blood pressure. PMID- 19298533 TI - Effects of sodium ferulate on human osteoarthritic chondrocytes and osteoarthritis in rats. AB - 1. Oxidation, inflammation and apoptosis are involved in the aetiology and pathology of osteoarthritis (OA). Sodium ferulate (SF) has been demonstrated to have anti-oxidant, anti-inflammatory and anti-apoptotic actions in cardiovascular, hepatic and diabetic disorders. These findings suggest that SF may have beneficial effects on OA. Therefore, present study investigated the effects of SF in an in vivo rat OA model, as well as in vitro in human OA chondrocytes. 2. Rats were divided into the following groups: (i) an untreated control group; (ii) papain-induced OA; (iii) OA rats treated with 0.1 or 0.5% SF; and (iv) normal rats injected with 0.5% SF intra-articularly. Human chondrocytes from OA patients were cultured before being stimulated with 2 ng/mL interleukin 1beta and subsequently treated with SF (125, 250 and 500 MUmol/L). The effects of SF were evaluated both in vivo and in vitro. 3. In OA rats, SF treatment dose dependently reversed pathological changes in OA cartilage, decreased BAX immunopositive chondrocytes and increased Bcl-2-immunopositive chondrocytes. Both in vivo and in vitro analyses demonstrated a significant decrease in matrix metalloproteinase-1 and an increase in tissue-specific inhibitor of metalloproteinase-1. In vitro, SF enhanced chondrocyte proliferation and decreased nitric oxide production and apoptosis. 4. The results demonstrate that SF dose-dependently suppresses pathological processes in both in vitro and in vivo OA models. Thus, SF could be a therapeutic strategy for the treatment of OA. PMID- 19298534 TI - Pharmacological changes in cellular Ca2+ homeostasis parallel initiation of atrial arrhythmogenesis in murine Langendorff-perfused hearts. AB - 1. Intracellular Ca(2+) overload has been associated with established atrial arrhythmogenesis. The present experiments went on to correlate acute initiation of atrial arrhythmogenesis in Langendorff-perfused mouse hearts with changes in Ca(2+) homeostasis in isolated atrial myocytes following pharmacological procedures that modified the storage or release of sarcoplasmic reticular (SR) Ca(2+) or inhibited entry of extracellular Ca(2+). 2. Caffeine (1 mmol/L) elicited diastolic Ca(2+) waves in regularly stimulated atrial myocytes immediately following addition. This was followed by a decline in the amplitude of the evoked transients and the disappearance of such diastolic events, suggesting partial SR Ca(2+) depletion. 3. Cyclopiazonic acid (CPA; 0.15 micromol/L) produced more gradual reductions in evoked Ca(2+) transients and abolished diastolic Ca(2+) events produced by the further addition of caffeine. 4. Nifedipine (0.5 micromol/L) produced immediate reductions in evoked Ca(2+) transients. Further addition of caffeine produced an immediate increase followed by a decline in the amplitude of the evoked Ca(2+) transients, without eliciting diastolic Ca(2+) events. 5. These findings correlated with changes in spontaneous and provoked atrial arrhythmogenecity in mouse isolated Langendorf-perfused hearts. Thus, caffeine was pro-arrhythmogenic immediately following but not > 5 min after application and both CPA and nifedipine pretreatment inhibited such arrhythmogenesis. 6. Together, these findings relate acute atrial arrhythmogenesis in intact hearts to diastolic Ca(2+) events in atrial myocytes that, in turn, depend upon a finite SR Ca(2+) store and diastolic Ca(2+) release following Ca(2+)-induced Ca(2+) release initiated by the entry of extracellular Ca(2+). PMID- 19298535 TI - Prevention of aortic elastic lamina defects by losartan in apolipoprotein(E) deficient mouse. AB - 1. In a previous study, we identified prevalent internal elastic lamina (IEL) defects in the aorta of hyperlipidaemic apolipoprotein E (ApoE)-deficient mice that are thought to provide a structural basis for the development of atherosclerosis and intimal thickening. In the present study, we examined the effects of losartan, an angiotensin AT1 receptor antagonist, on the development of IEL defects. 2. Male 18-week-old ApoE-deficient mice (maintained on a normal diet) were treated with losartan (3 or 30 mg/kg per day) for 10 weeks via the drinking water. The IEL defects were quantified histologically by measuring the continuity of the IEL within the inner curvature of the aortic arch. 3. In untreated animals, there was an age-dependent increase in IEL defects from 7.2 +/ 2.1% at 18 weeks to 13.8 +/- 4.0% at 28 weeks. Treatment with the high dose of losartan significantly prevented the development of IEL defects (4.7 +/- 1.3% at 28 weeks; P < 0.05 vs untreated). This effect was independent of changes in blood pressure or plasma lipid levels. Using quantitative real-time polymerase chain reaction, we found that the effects of losartan were not associated with changes in levels of matrix metalloproteinase (MMP)-2 and MMP-9, tissue inhibitor of matrix metalloproteinase-1 or inflammatory markers in the aorta. 4. The results suggest that the renin-angiotensin system may contribute to the development of aortic IEL defects in a blood pressure-independent manner. PMID- 19298536 TI - Fluoxetine protects against monocrotaline-induced pulmonary arterial hypertension: potential roles of induction of apoptosis and upregulation of Kv1.5 channels in rats. AB - 1. Suppressing apoptosis and downregulating K(+) channels in pulmonary artery smooth muscle cells (PASMC) have been implicated in the development of pulmonary vascular medial hypertrophy and pulmonary arterial hypertension (PAH). Previous studies have shown that selective serotonin re-uptake inhibitors (SSRIs) protected against PAH. The aim of the present study was to investigate the involvement of Kv1.5 channels and apoptosis in the protective effect of the SSRI fluoxetine against PAH. 2. Monocrotaline (MCT) was used to establish PAH in Wistar rats. Fluoxetine (2 and 10 mg/kg per day) was administered by gavage once a day for 3 weeks. Three weeks after the induction of PAH by MCT, pulmonary haemodynamic measurements and pulmonary artery morphological assessments were undertaken, along with detection of apoptosis and Kv1.5. 3. Fluoxetine (2 and 10 mg/kg per day) decreased pulmonary artery pressure, reduced the right ventricular index and inhibited the increase in medial wall thickness of pulmonary arteries in established PAH. Fluoxetine (10 mg/kg per day) reduced the expression of Bcl-2 and Bcl-xL protein, increased the expression of cleaved caspase 3 protein and enhanced the expression of Kv1.5 protein and mRNA in pulmonary arteries. Furthermore, fluoxetine (10 mg/kg per day) significantly suppressed proliferation and enhanced apoptosis of PASMC in MCT-induced PAH. 4. In conclusion, fluoxetine protects against MCT-induced PAH by suppressing PASMC proliferation, inducing PASMC apoptosis and upregulating Kv1.5 channels. PMID- 19298537 TI - Anti-apoptotic effect of heat shock protein 90 on hypoxia-mediated cardiomyocyte damage is mediated via the phosphatidylinositol 3-kinase/AKT pathway. AB - 1. Hypoxia-induced cardiomyocyte apoptosis contributes significantly to cardiac dysfunction following trauma, shock and burn injury. There is evidence that heat shock protein (HSP) 90 is anti-apoptotic in cardiomyocytes subjected to a variety of apoptotic stimuli. Because HSP90 acts as an upstream regulator of the serine/threonine protein kinase Akt survival pathway during cellular stress, we hypothesized that HSP90 exerts a cardioprotective effect via the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway. 2. Neonatal rat cardiomyocytes were subjected to normoxia or hypoxia in the absence or presence of the HSP90 inhibitor geldanamycin (1 MUg/mL). Cardiomyocyte apoptosis was assessed by release of lactate dehydrogenase (LDH), terminal deoxyribonucleotidyl transferase mediated dUTP-digoxigenin nick end-labelling (TUNEL) staining and caspase 3 activity. Expression of HSP90, Akt, Bad and cytochrome c release was determined by western blot analysis. 3. Following exposure of cells to hypoxia, HSP90 was markedly elevated in a time-dependent manner, reaching a peak at 6 h (eightfold increase). Geldanamycin significantly increased hypoxia-induced release of LDH by 114%, the percentage of apoptotic cardiomyocytes by 102% and caspase 3 activity by 78%. Pretreatment of cells with geldanamycin also suppressed phosphorylation of both Akt and its downstream target Bad, but promoted the mitochondrial release of cytochrome c. 4. In conclusion, HSP90 activity is enhanced in cardiomyocytes following hypoxic insult. The anti-apoptotic effect of HSP90 on cardiomyocytes subjected to hypoxia is mediated, at least in part, by the PI3-K/Akt pathway. Key words: apoptosis, cardiomyocyte, heart failure, heat shock protein 90, hypoxia, phosphatidylinositol 3-kinase/Akt signalling pathway, serine/threonine protein kinase Akt. PMID- 19298538 TI - Cardioprotective effect of L-glutamate in obese type 2 diabetic Zucker fatty rats. AB - 1. Because diabetic hearts have an increased threshold for cardioprotection by ischaemic preconditioning (IPC), we hypothesized that protection by L-glutamate during reperfusion is restricted in Type 2 diabetic hearts. Previously, we found that L-glutamate-mediated postischaemic cardioprotection mimics IPC. 2. Rat hearts were studied in a Langendorff preparation perfused with Krebs'-Henseleit solution and subjected to 40 min global no-flow ischaemia, followed by 120 min reperfusion. L-Glutamate (0, 15 and 30 mmol/L) was added to the perfusate during reperfusion of hearts from non-diabetic (Wistar-Kyoto) and diabetic (Zucker diabetic fatty (ZDF)) rats, studied at 16 weeks of age. The infarct size (IS)/area-at-risk (AAR) ratio was the primary end-point. Expression of L glutamate excitatory amino acid transporter (EAAT) 1 (mitochondrial) and EAAT3 (sarcolemmal) was determined by quantitative polymerase chain reaction and immunoblotting. 3. The ISS/AAR ratio did not differ between control hearts from Wistar-Kyoto and ZDF rats (0.52 +/- 0.03 and 0.51 +/- 0.04, respectively; P = 0.90). L-Glutamate (15 mmol/L) significantly reduced the IS/AAR ratio in non diabetic hearts, but not in diabetic hearts, compared with their respective controls. The higher concentration of L-glutamate (30 mmol/L) reduced infarct size in diabetic hearts to the same degree as in non-diabetic hearts (IS/AAR 0.35 +/- 0.03 (P = 0.002) and 0.34 +/- 0.03 (P = 0.004), respectively). The mitochondrial L-glutamate transporter EAAT1 was downregulated in hearts from ZDF rats at both the mRNA and protein levels (P < 0.0005 and P < 0.0001, respectively). However, there was no change in EAAT3 expression at the protein level. Myocardial L-glutamate content was increased by 43% in diabetic hearts (P < 0.0001). 4. Hearts from obese diabetic rats have an elevated threshold for metabolic postischaemic cardioprotection by L-glutamate. These findings may reflect underlying mechanisms of inherent resistance against additional cardioprotection in the diabetic heart. PMID- 19298539 TI - Hypoxia inducible factor-1 improves the actions of positive inotropic agents in stunned cardiac myocytes. AB - 1. In the present study, we tested hypothesis that upregulation of hypoxia inducible factor-1 (HIF-1) would improve the actions of positive inotropic agents in cardiac myocytes after simulated ischaemia-reperfusion (I/R). 2. Hypoxia inducible factor-1alpha was upregulated with deferoxamine (150 mg/kg per day for 2 days). Rabbit cardiac myocytes were subjected to simulated ischaemia (15 min, 95% N(2)-5% CO2) and reperfusion (re-oxygenation) and compared with control myocytes. Cell contraction and calcium transients were measured at baseline and after forskolin (10(-7) and 10(-6) mol/L) or ouabain (10(-5) and 10(-4) mol/L). 3. Under control conditions, high-dose forskolin and ouabain increased percentage shortening by 20 and 18%, respectively. Deferoxamine-treated control myocytes responded similarly. In stunned myocytes, forskolin and ouabain did not significantly increase shortening (increases of 8% and 9%, respectively). Deferoxamine restored the effects of forskolin (+26%) and ouabain (+28%) in stunning. The results for maximum shortening and relaxation rates were similar. The increased calcium transients caused by forskolin and ouabain were also depressed in stunned myocytes, but were maintained by HIF-1 upregulation. 4. These results suggest that simulated I/R impaired the functional and calcium transient effects of positive inotropic agents. Upregulation of HIF-1 protects cardiac myocyte function after I/R by maintaining calcium release. PMID- 19298540 TI - Chromium picolinate inhibits resistin secretion in insulin-resistant 3T3-L1 adipocytes via activation of amp-activated protein kinase. AB - 1. Chromium picolinate (CrPic) has been recommended as an alternative therapeutic regimen for Type 2 diabetes mellitus (T2DM). However, the molecular mechanism underlying the action of CrPic is poorly understood. 2. Using normal and insulin resistant 3T3-L1 adipocytes, we examined the effects of CrPic on the gene transcription and secretion of adiponectin and resistin. In addition, using immunoblotting, ELISA and real-time reverse transcription-polymerase chain reaction (RT-PCR), we investigated the effects of 10 nmol/L CrPic for 24 h on AMP activated protein kinase (AMPK) to determine whether this pathway contributed to the regulation of adiponectin and resistin expression and secretion. 3. Chromium picolinate did not modulate the expression of adiponectin and resistin; however, it did significantly inhibit the secretion of resistin, but not adiponectin, by normal and insulin-resistant 3T3-L1 adipocytes in vitro. Furthermore, although CrPic markedly elevated levels of phosphorylated AMPK and acetyl CoA carboxylase in 3T3-L1 adipocytes, it had no effect on the levels of AMPK alpha-1 and alpha-2 mRNA transcripts. Importantly, inhibition of AMPK by 2 h pretreatment of cells with 20 micromol/L compound C completely abolished the CrPic-induced suppression of resistin secretion. 4. In conclusion, the data suggest that CrPic inhibits resistin secretion via activation of AMPK in normal and insulin-resistant 3T3-L1 adipocytes. PMID- 19298541 TI - Effects of thoracic epidural anaesthesia on cardiac repolarization. AB - 1. Prolongation of the QT interval is associated with a risk of life-threatening cardiac arrhythmias. In the present study, we examined whether reversible blockade of preganglionic sympathetic fibres of the heart, achieved by thoracic epidural anaesthesia, affects cardiac repolarization and thus confers an anti arrhythmic effect. 2. Fifty-two men, aged between 28 and 65 years, were included in the study: 28 were patients scheduled for thoracic epidural anaesthesia (Group T) and 24 were patients scheduled for lumbar epidural anaesthesia (Group L). Epidural blockade was achieved with 0.5% isobaric bupivacaine solution. Measurements were taken from electrocardiogram fragments obtained before epidural anaesthesia and after detection of blockade (T1 or T8 segment sensory block in Groups T and L, respectively). Correction of the QT interval was calculated using Bazett's formula (QTcb), Fridericia's correction (QTcf) and the Framingham formula (QTcF). Transmural dispersion of repolarization (TDR) was determined using the Tpeak--Tend time, where Tpeak is the peak of the T wave and Tend is the end of the T wave. 3. Significant shortening of the QTcb interval and TDR was detected after blockade in Group T. However, there were no changes in the ECG parameters in Group L. 4. In conclusion, reversible blockade of preganglionic sympathetic fibres to the heart, achieved by thoracic epidural anaesthesia, results in a reduction in QTcb, as well as the TDR. These changes may explain the anti-arrhythmic action seen with central blockade. PMID- 19298542 TI - Effects of experimentally induced diabetes mellitus on pharmacologically and electrically elicited myometrial contractility. AB - 1. Diabetes is one of the most frequent complications of gestation, affecting approximately 7% of pregnancies. However, little is known about its effects on electrically and pharmacologically stimulated myometrial contractility. The aim of the present study was to investigate the consequences of streptozotocin (STZ) induced diabetes on: (i) electrical field stimulation (EFS)-evoked contraction of isolated uterine rings as a function of gestational age; and (ii) the uterotonic and tocolytic actions of alpha- and beta-adrenoceptor stimulation, respectively. The effects of oxytocin in late pregnancy were also investigated. 2. During pregnancy, EFS-evoked contractions of isolated uterine rings from intact rats declined, whereas isolated uterine rings from diabetic rats exhibited continuously low sensitivity to EFS. 3. In non-pregnant rats, diabetes resulted in increased noradrenaline-mediated contractility and a decreased relaxation response to terbutaline. At the mRNA level, diabetes enhanced the expression of alpha1B-adrenoceptors in non-pregnant rats from 14.65 to 18.39 MUg/mL (P < 0.05), whereas the expression of alpha1D-adrenoceptors decreased (from 42.87 to 35.67 MUg/mL; P < 0.05). During pregnancy, the responses to these sympathomimetics did not differ between diabetic and intact rats. 4. In late pregnancy (on Days 15 and 21), oxytocin caused greater maximum contractility of uterine rings from diabetic rats without affecting the EC(50). In addition, on Day 15 of pregnancy, the expression of oxytocin receptors in the myometrium of diabetic rats was higher than that in intact rats. 5. The results of the present study indicate that experimental diabetes facilitates gestation-induced denervation and increases myometrial sensitivity to oxytocin in late pregnancy. If similar mechanisms operate in humans, this could contribute to a tendency to premature uterine contractions in diabetes-complicated pregnancies. PMID- 19298543 TI - Effects of gender and age on paediatric headache. AB - The aim of this study was to evaluate the impact of gender and age on headache characteristics and disability. Headache characteristics were assessed at an initial visit to a paediatric specialty care centre and five follow-up visits. A total number of 4121 patients were evaluated. Fifty-eight per cent of the sample was female. Boys were younger at their first headache and initial visit. They more frequently described headache pain as squeezing and location as top of the head. Girls reported more frequent and longer headaches. Girls more often described headache pain as sharp and location as back of the head. Age accounted for more variance than gender in headache severity, duration, frequency and disability. Gender differences exist in headache characteristics. Age is also an important factor in the variability in characteristics and disability. Longitudinal studies are needed to describe further the natural history of headaches in childhood and compare outcome between genders. PMID- 19298544 TI - Association of von Willebrand factor activity with ACE I/D and MTHFR C677T polymorphisms in migraine. AB - Angiotensin-converting enzyme (ACE) insertion (I)/deletion (D) and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms are linked to endothelial dysfunction and to cerebral white matter lesions. Objectives of this study were to determine if ACE and MTHFR gene polymorphisms are associated with von Willebrand factor (vWF) activity, an endothelial dysfunction marker, and with a distinct headache phenotype. We enrolled 64 women (18-50 years old) with International Classification of Headache Disorders, 2nd edn migraine without aura (MoA) and 61 with aura (MA). Genotypic frequencies: ACE DD 35%, ID 42%, II 23%, and MTHFR TT 17%, CT 40%, CC 43%. Those with ACE DD genotype had higher levels of vWF activity (152%) compared with ID and II genotypes. Levels were highest (179%) with combined ACE DD and MTHFR TT genotypes. ACE DD was associated with higher headache frequency, and MTHFR TT was associated with MA. In migraine, vWF activity may be a marker of endothelial-mediated genetic risk for ischaemic conditions. PMID- 19298545 TI - Usefulness of a photophobia questionnaire in patients with migraine. AB - Photophobia is an important criterion for the diagnosis of migraine. However, several Asian epidemiological surveys about migraine have shown lesser prevalence of photophobia than that of Western studies. This discrepancy is probably caused by underestimation of photophobia due to inappropriate questioning of patients by physicians. To investigate this issue, we developed a questionnaire about photophobia and evaluated its usefulness in 103 patients with migraine. In phase 1 of the study, we found good repeatability of the questionnaire with a 0.826 kappa coefficient. In phase 2, the prevalence of photophobia from interviews and that from the questionnaire were compared. The prevalence of interview-documented photophobia was 51.5% and of questionnaire-documented photophobia 82.5% (P < 0.001). In phase 3, we attempted to make a short-form questionnaire with the same detection power of the questionnaire study. Two short-form questionnaires were identified as a useful method for detecting photophobia. The prevalence of photophobia could be underreported via interview, especially in Asian migraineurs. Using this questionnaire to test for photophobia, the diagnostic rate of photophobia and migraine could be improved. PMID- 19298546 TI - Methods of labeling skin surgical specimens. AB - BACKGROUND: Accurate pre-operative or intra-operative labeling of the skin is often necessary to mark exactly the surgical excision lines. Pre-operative "unsterile" permanent skin labeling systems are needed for example for vein and sentinel lymph node surgery; here the dyes must resist two surgical skin disinfection procedures. In contrast, excision borders are labeled during surgery using a "sterile" skin marking system. METHODS: Many commercial and non commercial pre- and intra-operative skin labeling systems are available, such as autologous patient blood, fluorescence triphenylmethane dyes and commercial skin markers. The available skin marking systems have specific advantages and disadvantages. We review the different labeling systems, offering guidelines to help choose a cost-effective system appropriate for a given surgical procedure. RESULTS: The Edding permanent markers 400 und 3000 are well suited for preoperative skin labeling and less expensive than commercial skin labeling systems. Autologous patient blood and eosin are well suited for intra-operative labeling and are most cost effective. Eosin Y is widely used and well suited for labeling of dark skin, bone, cartilage, and muscle tissue and spares the expense of expensive commercial skin markers. CONCLUSION: Knowledge of the many commercial and non-commercial pre- and intra-operative skin labeling systems and their advantages and disadvantages helps to reduce the use of relatively expensive commercial skin markers. PMID- 19298547 TI - Bacillary angiomatosis. AB - An infection with Bartonella henselae transmitted from domestic cats to humans by scratching normally leads to cat-scratch disease. When the human host has severe immunosuppression or HIV infection, the potentially life-threatening disease bacillary angiomatosis can develop. A 79-year-old man presented with livid erythematous, angioma-like skin lesions. We considered a cutaneous infiltrate from his known chronic lymphocytic leukemia, Merkel cell carcinoma, cutaneous metastases of internal tumors, cutaneous sarcoidosis, mycobacterial infection and even atypical herpes simplex infection. The correct diagnosis was proven histologically and by PCR. Because of increasing numbers of immunosuppressed and HIV-positive patients, as well as an infection rate of 13% for B. henselae in domestic cats in Germany, one must be alert to the presence of bacillary angiomatosis. PMID- 19298548 TI - Progress of multiple cutaneous and subcutaneous melanoma metastases of the face during imiquimod treatment. AB - A 68-year-old man developed a local recurrence with multiple cutaneous and subcutaneous nodules three months after excision of a primary malignant melanoma of the temple. Despite extensive surgery and adjuvant irradiation, another local recurrence occurred. Following further local progression during dacarbazine chemotherapy, topical treatment with imiquimod was begun and the chemotherapy was changed to fotemustine. During this treatment further local progression occurred and two months later regional lymph node and distant metastasis were detected. The patient died from his tumor disease eighteen months after the first diagnosis of malignant melanoma. PMID- 19298549 TI - Simulation of a mycological KOH preparation--e-learning as a practical dermatologic exercise in an undergraduate medical curriculum. AB - BACKGROUND: Mycological KOH preparation is one of the most popular practical laboratory skills in dermatology. The study addresses the question whether an interactive simulation program enhances knowledge of students about this procedure. METHODS: 166 students, 107 female and 59 male, participated. We separated our study in three phases: pretest, completing the simulation three times and post-test. In the pre- and post-test we recorded the number of correct steps of the mycological KOH preparation listed by the students. The full text feedback was explored by content analysis. RESULTS: In the pre-test the students listed an average of 3.1 +/- 2.2 correct steps, compared to 8.8 +/- 1.2 correct steps after completing the simulation (p < 0.001). Furthermore, the improvement was significant for each individual step. There were no significant differences between male and female students. In content analysis of the feedback, positive statements prevailed with 78.3%, compared to only 1.8% critical items. CONCLUSION: Our study shows that an interactive computer simulation program of mycological KOH preparation results in a significant learning effectiveness as far as recall of the correct procedural steps is concerned. Furthermore, subjective acceptance by students is high. PMID- 19298550 TI - Cerebral cortex development: From progenitors patterning to neocortical size during evolution. AB - The central nervous system is composed of thousands of distinct neurons that are assembled in a highly organized structure. In order to form functional neuronal networks, distinct classes of cells have to be generated in a precise number, in a spatial and temporal hierarchy and to be positioned at specific coordinates. An exquisite coordination of appropriate growth of competent territories and their patterning is required for regionalization and neurogenesis along both the anterior-posterior and dorso-ventral axis of the developing nervous system. The neocortex represents the brain territory that has undergone a major increase in its relative size during the course of mammalian evolution. In this review we will discuss how the fine tuning of growth and cell fate patterning plays a crucial role in the achievement of the final size of central nervous system structures and how divergence might have contributed to the surface increase of the cerebral cortex in mammals. In particular, we will describe how lack of precision might have been instrumental to neocortical evolution. PMID- 19298551 TI - Continuous neurogenesis in the adult brain. AB - Neurogenesis occurs throughout adulthood in the mammalian brain by coordinated proliferation and differentiation of adult neural stem cells. Newborn neurons are incorporated into the functional networks of both the olfactory bulb and the hippocampal dentate gyrus, suggesting significant roles of adult neurogenesis in brain functions. In this review, we discuss the recent findings about the integration mode of new neurons into the existing neural circuits. We further address the potential significance of adult neurogenesis in higher brain functions such as olfactory and spatial memory. PMID- 19298552 TI - From brain formation to plasticity: insights on Otx2 homeoprotein. AB - The shaping of neuronal circuits is essential during postnatal brain development. A window of neuronal remodeling by sensory experience typically occurs during a unique time in early life. The many types of behavior and perception, like human language, birdsong, hearing and vision are refined by experience during these distinct 'critical periods'. The onset of critical periods for vision is delayed in animals that remain in complete darkness from birth. It is then predicted that a 'messenger' within the visual pathway signals the amount of sensory experience that has occurred. Our recent results indicate that Otx2 homeoprotein, an essential morphogen for embryonic head formation, is reused later in life as this 'messenger' for critical period plasticity. The homeoprotein is stimulated by visual experience to propagate into the visual cortex, where it is internalized by GABAergic interneurons, especially Parvalbumin-positive cells (PV-cells). Otx2 promotes the maturation of PV-cells, consequently activating critical period onset in the visual cortex. Here, we discuss recent data that are beginning to illuminate the physiological function of non-cell autonomous homeoproteins, as well as the restriction of their transfer to PV-cells in vivo. PMID- 19298553 TI - Roles of planar cell polarity pathway genes for neural migration and differentiation. AB - Planar cell polarity (PCP) in epithelial cells is essential for the organization of tissues and their functions. The conserved non-canonical Wnt/PCP pathway regulates this process in both Drosophila and vertebrates. However, recent studies have revealed that a similar set of genes, which may not be related to PCP, regulates oriented cell movement during development. In the present review, recent findings on neural migration in zebrafish hindbrain and axonal guidance in mouse forebrain and spinal cord are discussed. Future analyses on defects in vertebrate PCP mutants will provide novel insights into the conserved and diverse roles of non-canonical Wnt/PCP pathway genes in vertebrate brain development. PMID- 19298554 TI - Development of the central and peripheral nervous systems in the lamprey. AB - Vertebrate brains are highly organized structures that show remarkable diversity throughout the animal groups. Among the vertebrates, the agnathan animals, which diverged from the gnathostomes early in the evolution of the vertebrates, occupy a key phylogenetic position in order to clarify the origin and evolution of the brain. We found that the lamprey brain has the basic molecular mechanisms necessary to form brain compartments. Conversely, the telencephalon and cerebellum display gnathostome-specific developmental mechanisms. We also propose that, in contrast to those of gnathostomes, the maxillary ramus of the trigeminal nerve and buccal ramus of the nerve on the anterior lateral line are not fused in the developing lamprey. Thus, the development of the central nervous system and the framework of the peripheral nerve around the oral region are thought to have improved in the course of the agnathan-gnathostome transition. PMID- 19298555 TI - Validation, evidence based medicine and Aristotle's wives. PMID- 19298556 TI - Successful left ventricular lead placement via an anomalous coronary sinus draining into the right atrial appendage. PMID- 19298557 TI - Pacing lead implantation without live fluoroscopy: feasibility of acute success in the live canine model. AB - INTRODUCTION: Fluoroscopic visualization for transvenous pacing lead placement necessitates lead shielding to minimize radiation exposure. An electromagnetic (EM) navigation system that integrates real-time intracardiac tracking within an anatomic navigation environment may provide an effective alternative for lead delivery that obviates live fluoroscopy. We assessed feasibility of pacing lead implantation with electromagnetic tracking guided solely by radiographic virtual navigation and compared this to fluoroscopy-guided implants in a canine model. METHODS: Seven mongrel dogs with normal hearts were randomized to 47 pacing lead placements in the right atrium (RA) or right ventricle (RV) guided by single plane fluoroscopy, or an experimental EM navigation system guided by registered fluoroscopic snapshots obtained before implant (EMN). Ability to achieve successful lead delivery acutely was assessed, and pacing parameters as well as fluoroscopy and implant times were measured. Means were compared using a paired t test. RESULTS: All lead delivery attempts were acutely successful. One atrial lead dislodged with EMN, resulting in 46 successful pacing attempts. There was no statistical difference in pacing parameters and time for lead placement between the approaches (EMN vs fluoroscopic navigation [mean +/- SD]: RA threshold 1.15 V +/- 0.98 V vs 1.95 V +/- 0.98 V [P = NS], RV threshold 1.18 V +/- 0.58 V vs 1.42 V +/- 0.63 V [P = NS], implant time 4:38 +/- 2:37 minutes vs 4:44 +/- 2:38 minutes [P = NS]). No live fluoroscopy was required for EMN implants. CONCLUSION: Pacing lead placement with an EM system guided by preprocedural fluoroscopic views is feasible and comparable to fluoroscopic navigation, and avoids the use of live fluoroscopy. PMID- 19298558 TI - A case of flecainide-induced hyponatremia. AB - We present a case of flecainide-induced hyponatremia in a 67-year-old woman who was treated for paroxysmal atrial tachycardia. She developed dizziness after starting flecainide and was found to be hyponatremic with a sodium level of 122 mmol/L (decreased from 136 mmol/L). Work-up failed to reveal other causes of hyponatremia. She was not on diuretics, laxatives, or herbal medications. After discontinuation of flecainide, her symptoms and sodium levels improved. Hyponatremia is a previously unrecognized entity as an adverse effect of flecainide. We will discuss the clinical presentation, lab findings, and a possible explanation for this patient's unusual reaction to flecainide. PMID- 19298559 TI - Potent antiarrhythmic effects of chronic amiodarone in canine pulmonary vein sleeve preparations. AB - OBJECTIVES: To examine the effects of chronic amiodarone on the electrophysiology of canine pulmonary vein (PV) sleeve preparations and left ventricular wedge preparation. BACKGROUND: Amiodarone is commonly used for the treatment of ventricular and supraventricular arrhythmias. Ectopic activity arising from the PV plays a prominent role in the development of atrial fibrillation (AF). METHODS: Standard microelectrode techniques were used to evaluate the electrophysiological characteristics of superfused PV sleeve (left superior or inferior) and arterially perfused left ventricular (LV) wedge preparations isolated from untreated and chronic amiodarone-treated dogs (amiodarone, 40 mg/kg daily for 6 weeks). RESULTS: In PV sleeves, chronic amiodarone (n = 6) induced a significant increase in action potential duration at 90% repolarization (APD90) and a significant use-dependent reduction in Vmax leading to 1:1 activation failure at long cycle lengths (basic cycle length of 124 +/- 15 ms in control vs 420 +/- 320 ms after chronic amiodarone [P < 0.01]). Diastolic threshold of excitation increased from 0.3 +/- 0.2 to 1.8 +/- 0.7 mA (P < 0.01). Delayed and late phase 3 early afterdepolarizations and triggered activity could be induced in PV sleeve preparations using acetylcholine (ACh, 1 microM), high calcium ([Ca2+]o = 5.4 mM), isoproterenol (Iso, 1 microM), or their combination in 6 of 6 untreated PV sleeves, but in only 1 of 5 chronic amiodarone-treated PV sleeve preparations. Vmax, conduction velocity, and 1:1 activation failure were much more affected in PV sleeves versus LV wedge preparations isolated from amiodarone treated animals. CONCLUSIONS: The results point to potent effects of chronic amiodarone to preferentially suppress arrhythmogenic substrates and triggers arising from the PV sleeves of the dog. PMID- 19298560 TI - Idiopathic focal ventricular arrhythmias originating from the anterior papillary muscle in the left ventricle. AB - INTRODUCTION: Focal ventricular arrhythmias (VAs) have been reported to arise from the posterior papillary muscle in the left ventricle (LV). We report a distinct subgroup of idiopathic VAs arising from the anterior papillary muscle (APM) in the LV. METHODS AND RESULTS: We studied 432 consecutive patients undergoing catheter ablation for VAs based on a focal mechanism. Six patients were identified with ventricular tachycardia (VT, n = 1) or premature ventricular contractions (PVCs, n = 5) with the earliest site of ventricular activation localized to the base (n = 3) or middle portion (n = 3) of the LV APM. No Purkinje potentials were recorded at the ablation site during sinus rhythm or the VAs. All patients had a normal baseline electrocardiogram and normal LV systolic function. The VAs exhibited a right bundle branch block (RBBB) and right inferior axis (RIA) QRS morphology in all patients. Oral verapamil and/or Na(+) channel blockers failed to control the VAs in 4 patients. VT was not inducible by programmed electrical stimulation in any of the patients. In 4 patients, radiofrequency current with an irrigated or conventional 8-mm-tip ablation catheter was required to achieve a lasting success. Two patients had recurrent PVCs after a conventional radiofrequency ablation with a 4-mm-tip ablation catheter had initially suppressed the arrhythmia. CONCLUSIONS: VAs may arise from the base or middle portion of the APM and are characterized by an RBBB and RIA QRS morphology and focal mechanism. Catheter ablation of APM VAs is typically challenging, and creation of a deep radiofrequency lesion may be necessary for long-term success. PMID- 19298561 TI - Utilization of retrograde right bundle branch block to differentiate atrioventricular nodal from accessory pathway conduction. AB - INTRODUCTION: Defining whether retrograde ventriculoatrial (V-A) conduction is via the AV node (AVN) or an accessory pathway (AP) is important during ablation procedures for supraventricular tachycardia (SVT). With the introduction of ventricular extrastimuli (VEST), retrograde right bundle branch block (RBBB) may occur, prolonging the V-H interval, but only when AV node conduction is present. We hypothesized that when AP conduction was present, the V-A interval would increase less than the V-H interval, whereas with retrograde nodal conduction, the V-A interval would increase at least as much as the V-H interval. METHODS AND RESULTS: We retrospectively reviewed the electrophysiological studies of patients undergoing ablation for AVN reentrant tachycardia (AVNRT) (55) or AVRT (50), for induction of retrograde RBBB during the introduction of VEST, and the change in the measured V-H and V-A intervals. Results were found to be reproducible between independent observers. Out of 105 patients, 84 had evidence of induced retrograde RBBB. The average V-H interval increase with induction of RBBB was 53.7 ms for patients with AVRT and 54.4 ms for patients with AVNRT (P = NS). The average V-A interval increase with induction of RBBB was 13.6 ms with AVRT and 70.1 ms with AVNRT (P < 0.001). All patients with a greater V-H than V-A interval change had AVRT, and those with a smaller had AVNRT. CONCLUSIONS: Induction of retrograde RBBB during VEST is common during an electrophysiological study for SVT. The relative change in the intervals during induction of RBBB accurately differentiates between retrograde AVN and AP conduction. PMID- 19298563 TI - Influence of left ventricular lead location on outcomes in the COMPANION study. AB - INTRODUCTION: There are no randomized controlled trial data that evaluate mortality and hospitalization rates in cardiac resynchronization therapy (CRT) recipients based on left ventricular (LV) lead location. We analyzed the event driven outcomes of mortality and hospitalization as well as functional outcomes including Functional Class, Quality-of-Life, and 6-minute walk distance in 1,520 patients enrolled in the COMPANION study of CRT versus optimal medical therapy. METHODS AND RESULTS: Over a mean follow-up after implantation of 16.2 months, patients randomized to CRT, regardless of lead location, experienced benefit compared with optimized pharmacologic therapy (OPT), with respect to all-cause mortality or heart failure hospitalization. All but a posterior location showed benefit with respect to the all-cause mortality or all-cause hospitalization outcome. Mortality benefit in CRT-D patients was indifferent to LV lead position. All functional outcomes including 6-minute walk distance, Quality-of-Life (QOL) and Functional Class improved with CRT, regardless of LV lead location. CONCLUSION: LV lead location was not a major determinant of multiple measures of response to CRT therapy in the COMPANION Trial. While acute data indicate that a left lateral LV lead location results in the most favorable hemodynamic response, these chronic data suggest that positioning an LV lead in an anterior rather than a lateral or posterior LV location has similar benefit. PMID- 19298564 TI - Unraveling the mechanism of a wide-complex tachycardia. PMID- 19298565 TI - Human model simulating right ventricular outflow tract tachycardia by high frequency stimulation in the left pulmonary artery: autonomics and idiopathic ventricular arrhythmias. AB - INTRODUCTION: Frequent monomorphic premature ventricular contractions (PVC) and/or ventricular tachycardia (VT) in patients with structurally normal heart usually arise from the right ventricular outflow tract (RVOT). An animal model simulating RVOT tachycardia by high-frequency stimulation (HFS) of the sympathetic input to the proximal pulmonary artery (PA) has been previously described. The aim of this study was to similarly induce RVOT tachycardia in humans. METHODS: In 9 patients with no history of ventricular arrhythmias, a circumferential catheter was placed in the left, main, and proximal PA to contact the endovascular circumference of the PA. A 50-ms train of HFS (200 Hz/0.3 ms pulse duration), coupled to atrial pacing, was applied at each bipolar pair of the circumferential catheter. The coupling interval was adjusted so that the 50 ms train occurred during the ventricular refractory period. RESULTS: In 6 out of 9 patients, HFS in the left PA during dobutamine infusion induced monomorphic PVCs and/or VT with left bundle branch block (LBBB) morphology and inferior axis at an average stimulation level of 12.5 +/- 2.7 V. HFS in the main PA and in the proximal PA did not induce any ventricular arrhythmias with the highest energy of 15 V in baseline state and during dobutamine infusion. HFS in the left PA was associated with hiccough in all patients. CONCLUSION: Stimulation of the sympathetic input to the left PA during dobutamine infusion induces PVCs and/or VT exhibiting LBBB-morphology and inferior axis, closely simulating clinical RVOT tachycardia in humans. PMID- 19298566 TI - Ten-year follow-up of cardiac sympathectomy in a young woman with catecholaminergic polymorphic ventricular tachycardia and an implantable cardioverter defibrillator. AB - Current recommendations for therapy of catecholaminergic ventricular tachycardia (CPVT) include beta blockade and implantable cardioverter defibrillators (ICDs). Patients may experience recurrent arrhythmias, ICD shocks and, rarely, sudden death despite optimal medical therapy. We report a young woman with CPVT who received frequent ICD shocks despite beta blockade, who subsequently underwent cardiac sympathectomy with a dramatic reduction in shocks over 10 years of follow up. PMID- 19298567 TI - The prognostic significance of bundle branch block in high-risk chronic stable vascular disease patients: a report from the HOPE trial. AB - OBJECTIVE: The prognostic significance of left and right bundle branch block (LBBB and RRBB) in patients with chronic stable cardiovascular (CV) disease is not well characterized and was evaluated in the Heart Outcomes Prevention Evaluation (HOPE) study cohort. DESIGN: Observational analysis of data prospectively collected in the HOPE trial. SETTING AND PATIENTS: HOPE was a multicenter, international trial, which evaluated ramipril and vitamin E in 9,541 patients aged > or = 55 years with CV disease or diabetes with > or = 1 CV risk factor(s) but without heart failure (HF) or known left ventricular systolic dysfunction. Follow-up extended for a median of 4.5 years. Electrocardiograms were obtained at baseline in all study participants and were read centrally. MAIN OUTCOME MEASURES: Major CV events (defined as CV death, myocardial infarction, or stroke), heart failure, CV death, all-cause death, and sudden death. RESULTS: Baseline LBBB was present in 246 (2.6%) patients and was associated with increased risk for major CV events (HR = 1.54; 95% CI, 1.18-2.02), CV death (HR 2.29; 95% CI, 1.63-3.20), heart failure (HR 2.99; 95% CI, 2.31-3.87), sudden death (HR 3.17; 95% CI, 2.13-4.73), and all-cause death (HR = 2.10; 95% CI, 1.59 2.77). In multivariate models, LBBB remained an independent predictor of heart failure, sudden death, CV death, and all-cause death (P < or = 0.002 for all). Baseline RBBB was present in 428 (4.5%) of patients and was not associated with increased CV risk. CONCLUSIONS: In patients with stable chronic CV disease, LBBB but not RBBB is an independent predictor of heart failure, sudden death, CV death, and all-cause death. PMID- 19298568 TI - Effect of electroanatomically guided versus conventional catheter ablation of typical atrial flutter on the fluoroscopy time and resource use: a prospective randomized multicenter study. AB - AIMS: Radiofrequency catheter ablation of typical atrial flutter is one of the most frequent indications for catheter ablation in electrophysiology laboratories today. Clinical utility of electroanatomic mapping systems on treatment results and resource utilization compared with conventional ablation has not been systematically investigated in a prospective multicenter study. METHODS AND RESULTS: In this prospective, randomized multicenter study, the results of catheter ablation to cure typical atrial flutter using conventional ablation strategy were compared with electroanatomically guided mapping and ablation (Carto). Primary endpoints of the study were procedure duration and fluoroscopy exposure time, secondary endpoints were acute success rate, recurrence rate, and resource utilization. A total of 210 patients (169 men, 41 women, mean age 63 +/- 10 years) with documented typical atrial flutter were included in the study. Acute ablation success, that is, demonstration of bidirectional isthmus block, was achieved in 99 of 105 patients (94%) in the electroanatomically guided ablation group and in 102 of 105 patients (97%) in the conventional ablation group (P > 0.05). Total procedure duration was comparable between both study groups (99 +/- 57 minutes vs 88 +/- 54 minutes, P > 0.05). Fluoroscopy exposure time was significantly shorter in the electroanatomically guided ablation group (7.7 +/- 7.3 minutes vs 14.8 +/- 11.9 minutes; P < 0.05). Total recurrence rate of typical atrial flutter at 6 months of follow-up was comparable between the 2 groups (respectively for the CARTO and conventional group 6.6% vs 5.7%, P > 0.05). The material costs per procedure in the electroanatomically guided and conventional groups (NaviStar DS vs Celsius DS) was 3035 euro (USD 3,870) and 2133 euro (USD 2,720), respectively. CONCLUSIONS: This multicenter study documented that cavotricuspid isthmus ablation to cure typical atrial flutter was highly effective and safe, both in the conventional and the electroanatomically guided ablation group. The use of electroanatomical mapping system significantly reduced the fluoroscopy exposure time by almost 50%, however, at the expense of increased cost of the procedure. PMID- 19298569 TI - Heart rate turbulence as a predictor of cardiac mortality and arrhythmic events in patients with dilated cardiomyopathy: a prospective study. AB - BACKGROUND: Few studies have described the clinical usefulness of heart rate turbulence (HRT), an autonomic predictor of mortality, in stratifying patients with dilated cardiomyopathy (DCM) at risk of cardiac mortality and arrhythmic events. We prospectively assessed the utility of HRT for risk stratification in patients with ischemic or nonischemic DCM. METHODS: We enrolled 375 consecutive patients with DCM including ischemic (n = 241) and nonischemic causes (n = 134). HRT was measured using an algorithm based on routine 24-hour Holter electrocardiograms, assessing 2 parameters: turbulence onset (TO) and turbulence slope (TS). HRT was considered positive when both TO was > or = 0% and TS was < or = 2.5 ms/R-R interval. The primary endpoint was defined as cardiac mortality and the secondary endpoint as occurrence of hemodynamically stable sustained ventricular tachyarrhythmias. RESULTS: Of patients enrolled, 83 patients (22.1%) were not utilized for HRT assessment because there were too few ventricular premature beats, or for other reasons. Eighty-one of 292 patients (27.7%) were HRT-positive. During follow-up of 445 +/- 216 days, 30 patients (10.3%) reached the primary endpoint and 17 patients, the secondary endpoint. The hazard ratio (HR) of patients with an HRT-positive outcome was 6.4 (95%CI, 3.0-14.1; P < 0.0001) for the primary endpoint and 5.1 (95%CI, 2.8-9.3; P < 0.0001) for combined endpoints. On subanalysis, HRT positivity was significantly associated in both the ischemic and nonischemic DCM patients with both the primary endpoint (HR = 4.9, P = 0.0006 and HR = 12.3, P = 0.002, respectively) and with combined endpoints. CONCLUSIONS: HRT is a powerful risk stratification marker for cardiac mortality and arrhythmic events in patients with DCM whether ischemia is present or not. PMID- 19298570 TI - Ranolazine exerts potent effects on atrial electrical properties and abbreviates atrial fibrillation duration in the intact porcine heart. AB - INTRODUCTION: In vitro studies and ambulatory ECG recordings from the MERLIN TIMI 36 clinical trial suggest that the novel antianginal agent ranolazine may have the potential to suppress atrial arrhythmias. However, there are no reports of effects of ranolazine on atrial electrophysiologic properties in large intact animals. METHODS AND RESULTS: In 12 closed-chest anesthetized pigs, effects of intravenous ranolazine (approximately 9 microM plasma concentration) on multisite atrial effective refractory period (ERP), conduction time (CT), and duration and inducibility of atrial fibrillation (AF) initiated by intrapericardial acetylcholine were investigated. Ranolazine increased ERP by a median of 45 ms (interquartile range 29-50 ms; P < 0.05, n = 6) in right and left atria compared to control at pacing cycle length (PCL) of 400 ms. However, ERP increased by only 28 (24-34) ms in right ventricle (P < 0.01, n = 6). Ranolazine increased atrial CT from 89 (71-109) ms to 98 (86-121) ms (P = 0.04, n = 6) at PCL of 400 ms. Ranolazine decreased AF duration from 894 (811-1220) seconds to 621 (549-761) seconds (P = 0.03, n = 6). AF was reinducible in 1 of 6 animals after termination with ranolazine compared with all 6 animals during control period (P = 0.07). Dominant frequency (DF) of AF was reduced by ranolazine in left atrium from 11.7 (10.7-20.5) Hz to 7.6 (2.9-8.8) Hz (P = 0.02, n = 6). CONCLUSIONS: Ranolazine, at therapeutic doses, increased atrial ERP to greater extent than ventricular ERP and prolonged atrial CT in a frequency-dependent manner in the porcine heart. AF duration and DF were also reduced by ranolazine. Potential role of ranolazine in AF management merits further investigation. PMID- 19298571 TI - Robotic catheter ablation of ventricular tachycardia in a patient with congenital heart disease and Rastelli repair. AB - Robotically assisted catheter ablation has been proven feasible in patients with a variety of atrial arrhythmias. The potential to provide improved catheter tip maneuvering and stability potentially makes it ideal for complex ablation procedures. We present the case of a patient with complex congenital heart disease with previous Rastelli repair and recurrent ventricular tachycardia (VT) who underwent robotically assisted mapping and ablation for right ventricular VT, utilizing substrate mapping techniques. PMID- 19298572 TI - Long-term study of fluticasone propionate aqueous nasal spray in acute and maintenance therapy of nasal polyposis. AB - BACKGROUND: Topical steroids are first-line medication to control nasal polyposis (NP), a disease with long-term clinical course. OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of fluticasone propionate aqueous nasal spray (FPANS) 200 microg twice a day (bd) after 1 month of treatment, and to compare FPANS 200 microg bd and FPANS 200 microg once a day (od) in maintenance and long-term treatment. METHODS: Double-blind, placebo-controlled, 8-month study with three treatment periods (1-month acute period followed with 1-month maintenance period and 6-month follow-up period) was carried out. Group 1 received FPANS 200 microg bd, during acute, maintenance and follow-up periods, Group 2 received FPANS 200 microg bd during acute period and FPANS 200 microg od during maintenance and follow-up periods, and Group 3 received placebo during acute and maintenance periods and FPANS 200 microg bd during follow-up period. Endpoints were change from baseline in clinic peak nasal inspiratory flow (PNIF), domiciliary evening PNIF, intensity of symptoms and polyposis grade. RESULTS: After acute period and maintenance periods, FPANS 200 microg bd was significantly more effective than placebo on all endpoints and more effective than FPANS 200 microg od after 1-month maintenance period on clinic PNIF, evening PNIF, obstruction, percentage of days with no sense of smell and percentage of nights with no disturbances. The two doses were similar on other endpoints. After the 6 month follow-up period, there was no difference between the two doses of FPANS at all efficacy endpoints. The safety profile of FPANS did not highlight any new or unanticipated adverse events. CONCLUSION: The study demonstrated the efficacy of FPANS 200 microg bd in acute treatment and FPANS 200 microg od as a sufficient dose to maintain a long-term efficacy in the treatment for NP. PMID- 19298573 TI - A systematic review of studies validating the Edinburgh Postnatal Depression Scale in antepartum and postpartum women. AB - OBJECTIVE: The Edinburgh Postnatal Depression Scale (EPDS) is the most widely used screening tool for postpartum depression (PPD). We systematically reviewed the published evidence on its validity in detecting PPD and antepartum depression (APD) up to July 2008. METHOD: Systematic review of validation studies of the EPDS included 1987-2008. Cut-off points of 9/10 for possible PPD, 12/13 for probable PPD and 14/15 for APD were used. RESULTS: Thirty-seven studies met the inclusion criteria. Sensitivity and specificity of cut-off points showed marked heterogeneity between different studies. Sensitivity results ranged from 34 to 100% and specificity from 44 to 100%. Positive likelihood ratios ranged from 1.61 to 78. CONCLUSION: Heterogeneity among study findings may be due to differences in study methodology, language and diagnostic interview/criteria used. Therefore, the results of different studies may not be directly comparable and the EPDS may not be an equally valid screening tool across all settings and contexts. PMID- 19298574 TI - Detection of periapical bone defects in human jaws using cone beam computed tomography and intraoral radiography. AB - AIM: To compare the diagnostic accuracy of intraoral digital periapical radiography with that of cone beam computed tomography (CBCT) for the detection of artificial periapical bone defects in dry human jaws. METHODOLOGY: Small and large artificial periapical lesions were prepared in the periapical region of the distal root of six molar teeth in human mandibles. Scans and radiographs were taken with a charged couple device (CCD) digital radiography system and a CBCT scanner before and after each periapical lesion had been created. Sensitivity, specificity, positive predictive values, negative predictive values and Receiver Operator Characteristic (ROC) curves as well as the reproducibility of each technique were determined. RESULTS: The overall sensitivity was 0.248 and 1.0 for intraoral radiography and CBCT respectively, i.e. these techniques correctly identified periapical lesions in 24.8% and 100% of cases, respectively. Both imaging techniques had specificity values of 1.0. The ROC Az values were 0.791 and 1.000 for intraoral radiography and CBCT, respectively. CONCLUSIONS: With intraoral radiography, external factors (i.e. anatomical noise and poor irradiation geometry), which are not in the clinician's control, hinder the detection of periapical lesions. CBCT removes these external factors. In addition, it allows the clinician to select the most relevant views of the area of interest resulting in improved detection of the presence and absence of artificial periapical lesions. PMID- 19298575 TI - Canal transportation and centring ability of RaCe rotary instruments. AB - AIM: Evaluate, through computerized tomography, canal transportation and centring ability of RaCe rotary instruments after preparation of mesiobuccal root canals in maxillary molar teeth. METHODOLOGY: Twenty-seven teeth were submitted to three cone beam tomographic analyses, one preoperatively, and two after preparation with file size 35, .02 taper and size 50, .02 taper. Canal transportation and centring ability were measured with reference to the distance between the noninstrumented portion of the root canals and the mesial and distal periphery of the root, compared with images obtained after the preparation with size 35 and 50 instruments. RESULTS: Canal transportation after preparation with the size 35 file was 0.030 +/- 0.253 mm and after the size 50 file was 0.057 +/- 0.317 mm. The centring ratio values after preparation with the size 35 file was 0.42 +/- 0.32 and after the size 50 file was 0.54 +/- 0.29, with no significant statistical difference between the groups. CONCLUSIONS: RaCe instruments allowed the preparation of curved root canals with preparation diameters larger than those normally used with minimal canal transportation and adequate centring ability. PMID- 19298576 TI - New dimensions in endodontic imaging: Part 2. Cone beam computed tomography. AB - Cone beam computed tomography (CBCT) has been specifically designed to produce undistorted three-dimensional information of the maxillofacial skeleton, including the teeth and their surrounding tissues with a significantly lower effective radiation dose compared with conventional computed tomography (CT). Periapical disease may be detected sooner using CBCT compared with periapical views and the true size, extent, nature and position of periapical and resorptive lesions can be assessed. Root fractures, root canal anatomy and the nature of the alveolar bone topography around teeth may be assessed. The aim of this paper is to review current literature on the applications and limitations of CBCT in the management of endodontic problems. PMID- 19298577 TI - New dimensions in endodontic imaging: part 1. Conventional and alternative radiographic systems. AB - Conventional radiographs used for the management of endodontic problems yield limited information because of the two-dimensional nature of images produced, geometric distortion and anatomical noise. These factors often act in combination. This review paper assesses the limitations of periapical radiographs and seeks to clarify three-dimensional imaging techniques that have been suggested as adjuncts to conventional radiographs. These include tuned aperture computed tomography, magnetic resonance imaging, ultrasound, computed tomography and cone beam computed tomography (CBCT). Of these techniques, CBCT appears to be an effective and safe way to overcome some of the problems associated with conventional radiographs. PMID- 19298578 TI - A stab in the dark: an unusual anorectal knife wound. PMID- 19298579 TI - Extent of prevalence and size of flat neoplasms in a heterogeneous population undergoing routine colorectal cancer screening. AB - BACKGROUND: The importance of identifying flat colorectal neoplasms is increasingly appreciated, although the extent of prevalence of these lesions in a general population is not known. OBJECTIVE: To determine the extent of prevalence of flat neoplasms in a diverse population undergoing routine endoscopic screening for colorectal cancer. DESIGN: Patients referred to the Colorectal Cancer Screening Clinic over a 12-month period (n = 642). RESULTS: The patient population was 56% African American and 21% Caucasian; with a mean age of 59 + or - 9 years. Flat neoplasms were detected in 5.5% of all patients, similar to that reported elsewhere, with extent of prevalence being similar regardless of gender or race. Average size of flat neoplasms was of 2.8 + or - 2.3 mm (range 1-20 mm). However, there was no evidence of advanced pathology in any of the flat neoplasms identified. CONCLUSIONS: Flat neoplasms are common but may not be associated with advanced pathology in a population undergoing routine screening. PMID- 19298580 TI - Review article: dual delayed release formulation of dexlansoprazole MR, a novel approach to overcome the limitations of conventional single release proton pump inhibitor therapy. AB - BACKGROUND: Proton pump inhibitors (PPIs) provide the most effective pharmacotherapy for treating acid-related disorders. However, PPIs do not completely control acid over 24 h with once-daily dosing. AIMS: To discuss limitations inherent in the pharmacokinetics (PK) and pharmacodynamics of conventional PPI formulations, which provide a single drug release. Also, to consider approaches to extending the duration of acid suppression focusing on dexlansoprazole MR, a PPI with a novel Dual Delayed Release (DDR) formulation. METHOD: We reviewed the available literature regarding marketed and investigational PPIs. RESULTS: Non-standard dosing of currently marketed PPIs has produced incremental advances in acid control. Multiple approaches are being evaluated to enhance acid suppression with PPIs. Dexlansoprazole MR is a DDR formulation of dexlansoprazole, an enantiomer of lansoprazole, with two distinct drug release periods to prolong the plasma dexlansoprazole concentration-time profile and extend duration of acid suppression. Clinical studies show that dexlansoprazole MR produces a dual-peak PK profile that maintains therapeutic plasma drug concentrations longer than lansoprazole, with a single-peak PK profile, and increases the percentage of time that intragastric pH >4. CONCLUSIONS: Novel drug delivery platforms, including the dexlansoprazole MR DDR formulation, may improve acid suppression and offer benefits over conventional single release PPI formulations. PMID- 19298581 TI - Clinical trial: factors associated with freedom from relapse of heartburn in patients with healed reflux oesophagitis--results from the maintenance phase of the EXPO study. AB - BACKGROUND: Ability to predict freedom from heartburn relapse during maintenance therapy for healed reflux oesophagitis may facilitate optimal treatment choices for individual patients. AIM: To determine factors predicting freedom from heartburn relapse during maintenance proton pump inhibitor therapy in patients with healed reflux oesophagitis. METHODS: This post-hoc analysis used data from the maintenance phase of the EXPO study (AstraZeneca study code: SH-NEG-0008); 2766 patients with healed reflux oesophagitis and resolved heartburn received once-daily esomeprazole 20 mg or pantoprazole 20 mg for 6 months. Multiple logistic regression analysis determined factors associated with freedom from heartburn relapse. RESULTS: Heartburn relapse rates were lower with esomeprazole than pantoprazole in all subgroups analysed. Esomeprazole treatment was the factor most strongly associated with freedom from heartburn relapse (odds ratio 2.08; P < 0.0001). Other factors significantly associated with freedom from heartburn relapse were Helicobacter pylori infection, greater age, non-obesity, absence of epigastric pain at baseline, pre-treatment nonsevere heartburn and GERD symptom duration < or =5 years. CONCLUSIONS: Several factors predict freedom from heartburn relapse during maintenance proton pump inhibitor therapy for healed reflux oesophagitis, the strongest being choice of proton pump inhibitor. These findings outline the importance of optimizing acid control and identifying predictors of relapse for effective long-term symptom management in reflux oesophagitis patients. PMID- 19298582 TI - Risk of upper gastrointestinal bleeding with oral bisphosphonates and non steroidal anti-inflammatory drugs: a case-control study. AB - BACKGROUND: Gastrointestinal injuries including gastric ulcers have been reported with oral bisphosphonate therapy. However, the risk of the more serious upper gastrointestinal bleeding (UGB) especially in the community setting with these drugs remains unknown. Similarly, the risk of UGB among users of both bisphosphonates and non steroidal anti-inflammatory drugs (NSAIDs) in the community is also unknown. AIM: To explore the risk of more serious UGB among users of bisphosphonates and the risk of UGB among users of both bisphosphonates and NSAIDs in the community. METHODS: We conducted a case-control study within a cohort of Quebec residents who had received a revascularization procedure from 1995 to 2004. Cohort members were followed up from the date of their first procedure until the earliest of: (1) study outcome, (2) date of death or (3) end of health care coverage. Cases were defined as those with the first diagnosis of a UGB. For each case, 20 controls were selected and matched to the cases by index date, age and cohort entry. Adjusted odds ratios for current use of bisphosphonates, NSAIDs and co-therapy of both drugs were computed. RESULTS: Within the initial cohort, 3253 incident cases of UGBs and corresponding 65 060 matched controls were identified. The adjusted odds ratio (OR) for UGB by current users of bisphosphonates was 1.01 (95% CI, 0.72-1.43). Current NSAID use was associated with an increased risk of UGB OR = 1.75; 95% CI, 1.53-1.99. The OR for use of bisphosphonates and NSAIDs was elevated OR = 2.00; 95% CI, 1.12-3.57. This risk was still elevated for users of bisphosphonates and COX-2 inhibitors [OR = 2.38 (95% CI, 1.26-4.50)]. CONCLUSION: We found no evidence of an increase in the risk of UGB among current users of bisphosphonates. The risk of combined NSAID and bisphosphonate therapy was increased, but this risk was not higher than the risk for NSAID users alone. PMID- 19298583 TI - Review article: future therapies for management of metastatic gastroenteropancreatic neuroendocrine tumours. AB - BACKGROUND: Gastroenteropancreatic neuroendocrine tumours (GEP NETs) are relatively uncommon tumours that occur anywhere within the gastrointestinal tract. The prevalence of GEP NETs is estimated to be 35 per 100 000 population. Patients often present with metastatic disease and consequently, palliative treatments form the mainstay of therapy. AIM: To review the current and novel therapeutic options for management of GEP NETs. METHODS: Searches for all studies related to GEP NETs, NETs and carcinoid tumours in Medline and abstracts from international meetings. RESULTS: Somatostatin analogues remain the first line therapy for management of symptoms of GEP NETs and may have anti-proliferative action. New somatostatin analogues with different somatostatin receptor affinity have been developed. Radionuclide peptide receptor therapy is established in patients with positive somatostatin scintigraphy. A number of new agents and targeted therapies are currently being evaluated in a phase I and II studies and these include angiogenic inhibitors, mammalian target of rapamycin inhibitors and immune therapies. CONCLUSIONS: A number of nonsurgical therapies are available for management of gastroenteropancreatic neuroendocrine tumours. It is hoped, the development of some of these promising novel therapies will expand the therapeutic armamentarium. PMID- 19298584 TI - Increased serum phosphate levels and calcium fluxes are seen in smaller individuals after a single dose of sodium phosphate colon cleansing solution: a pharmacokinetic analysis. AB - BACKGROUND: Sodium phosphate containing colonoscopy preparations may cause electrolyte disturbances and calcium-phosphate nephropathy. Decreased body weight is an unexplored risk factor for complications with sodium phosphate ingestion. AIM: To perform a pharmacokinetic analysis of a single dose of Fleet Phospho-Soda in smaller and larger individuals. METHODS: Seven subjects weighing <55 kg (Group I) and six weighing >100 kg (Group II) consumed 45 mL Fleet Phospho-Soda. Serum electrolytes were measured. Hydration was closely maintained by monitoring weight, fluid intake and total body water. RESULTS: Marked increases in serum phosphate were seen in Group I compared to Group II. For example, mean serum phosphate at 120 min was 7.8 +/- 0.5 mg/dL in Group I and 5.1 +/- 0.8 mg/dL in Group II (P < 0.001). Normalized area under the phosphate vs. time curve for Group I was 1120 +/- 190 mg/dL*min and 685 +/- 136 mg/dL*min for Group II (P < 0.001). Twelve-hour urine calcium was lower in Group I (16.4 +/- 7.6 mg) than in Group II (39.2 +/- 7.8 mg, P < 0.001). CONCLUSIONS: Increased serum phosphate occurs in smaller individuals after ingestion of sodium phosphate preparations, even with strict attention to fluid intake. Smaller body weight poses a potential risk for calcium-phosphate nephropathy. PMID- 19298585 TI - Ghrelin receptor agonist (TZP-101) accelerates gastric emptying in adults with diabetes and symptomatic gastroparesis. AB - BACKGROUND: TZP-101 is a synthetic, selective ghrelin agonist in development for gastroparesis. AIM: To assess safety and effects of TZP-101 in diabetes patients with symptomatic gastroparesis. METHODS: Adults with type 1 or type 2 diabetes mellitus received placebo and TZP-101 (80, 160, 320 or 600 microg/kg) infusions in a cross-over manner following a radiolabelled meal. Blood glucose levels were stabilized using a hyperinsulinemic-euglycemic clamp. Primary endpoints were gastric half emptying and latency times. Secondary measures included assessment of gastroparesis symptoms and endocrine responses. RESULTS: Ten patients with type 1 (n = 7) or 2 (n = 3) diabetes, moderate-to-severe gastroparesis symptoms and > or =29% retention 4 h after a radiolabelled solid meal were enrolled. TZP 101 produced significant reductions in solid meal half-emptying (20%, P = 0.043) and latency (34%, P = 0.037) times vs. placebo. Reductions in overall postmeal symptom intensity (24%) and postprandial fullness (37%) following TZP-101 infusion were not statistically significant. Most adverse events were mild and self-limiting and there were no identifiable differences in numbers or types of adverse events between TZP-101 and placebo. CONCLUSIONS: This proof-of-concept study demonstrates that the ghrelin agonist TZP-101 is well-tolerated in diabetes patients with moderate-to-severe chronic gastroparesis and shows statistically significant improvements in gastric emptying. PMID- 19298586 TI - Aberrant expression of microRNA biosynthetic pathway components is a common feature of haematological malignancy. PMID- 19298587 TI - Prognostic factors influencing survival in solitary plasmacytoma. PMID- 19298588 TI - Soluble CD40 ligand and endothelial dysfunction in aspirin-treated polycythaemia vera patients. PMID- 19298589 TI - MicroRNA-27 enhances differentiation of myeloblasts into granulocytes by post transcriptionally downregulating Runx1. AB - We investigated the regulation of the transcription factor Runx1 by microRNA (miR)-27 and the resulting effects upon the differentiation of myeloblasts into granulocytes. When 32D.cl3 cell differentiation was induced using granulocyte colony-stimulating factor (CSF3), Runx1 transcription was moderately downregulated, while Runx1 protein levels were completely inhibited, suggesting an involvement of post-transcriptional regulation. Simultaneously, levels of miR 27 and its precursor increased substantially. Reporter assays revealed that miR 27 targets the 3'UTR of the Runx1 transcript. Furthermore, introduction of pre miR-27 alone into 32D.cl3 cells resulted in downregulation of Runx1 protein, thereby allowing the cell differentiation even in the absence of CSF3. Conversely, transduction of anti-miR-27 caused upregulation of Runx1 protein, thereby antagonizing the CSF3-mediated granulocyte differentiation. Finally, the CSF3-induced transcription factor C/EBPalpha enhanced transcription of a host gene of miR-27, C9orf3, via activation of its promoter. Thus, miR-27 enhances differentiation of myeloblasts into granulocytes via post-transcriptional downregulation of Runx1. PMID- 19298590 TI - Methotrexate and aminopterin exhibit similar in vitro and in vivo preclinical activity against acute lymphoblastic leukaemia and lymphoma. AB - Due to the development of neurological toxicity and resistance to methotrexate (MTX), other antifolates have been evaluated for its potential replacement in the treatment of childhood acute lymphoblastic leukaemia (ALL). Aminopterin (AMT) has been suggested to provide clinical advantages over MTX and other antifolates. AMT activity, compared with MTX, was evaluated in ALL and lymphoma preclinical models. The minimum survival fraction at the range of concentrations tested was lower with AMT than with MTX in 3 out of 15 cell lines. Both AMT and MTX significantly extended the event-free survival of mice bearing 3 out of 4 xenografts with equivalent activity. PMID- 19298591 TI - Notoriety to respectability: a short history of arsenic prior to its present day use in haematology. AB - This paper looks at arsenic, and in particular the trioxide, from the days of the ancient Chinese, Egyptians, Greeks and Romans, through the 17th-20th centuries to its adoption by today's haematologists. It looks at its commercial and medical uses, past and present, its notoriety as a poison, it's reputation as a 'tonic' and therapeutic agent, many of the famous people associated with it including Thomas Fowler, William Withering and Robert Christison, and the promise an 18th century panacea now offers 21st century patients under the care of today's haematologists and tomorrow's oncologists. PMID- 19298592 TI - The imbalance between Survivin and Bim mediates tumour growth and correlates with poor survival in patients with multiple myeloma. AB - Survivin is selectively expressed in most of common human cancers and is now viewed as a potent modulator of the cell death/proliferation balance in tumour cells. We previously found that myeloma cells expressed high levels of Survivin protein in correlation with disease progression and that Survivin knock-down by RNA interference decreased myeloma cell growth. We now demonstrate that Survivin overexpression promotes the proliferation and survival of human myeloma cells both in vitro and in vivo in the absence of their major growth factor, interleukin 6. Of particular interest, this effect correlates with the down regulation of Bim, a critical BH3-only cell death activator during cytokine deprivation, mainly at transcriptional level. The tight link between Survivin and Bim expression, reported for the first time here in myeloma cells and in other cell lines, is further confirmed in a panel of newly diagnosed patients with myeloma, and BIRC5 is validated as a gene significantly associated with short survival in these patients. Altogether, our findings provide evidence that Survivin directly contributes to malignant progression of myeloma and strongly suggest that targeting Survivin may disrupt the delicate balance controlling cell survival and proliferation, opening new avenues for the therapy of this still difficult-to-treat cancer. PMID- 19298593 TI - Therapeutic opportunities for counteracting apoptosis resistance in childhood leukaemia. AB - Evasion of apoptosis is a hallmark of human cancers, for example in haematological malignancies. Apoptosis is an intrinsic cell death program that is crucial in maintaining tissue homeostasis, for example in the haematopoietic system where there is a high turnover rate of cells. As a result, a decrease in the rate of apoptosis as well as an increase in proliferation favours tumorigenesis as well as tumour progression. Further, the anti-leukaemic action of current treatment approaches, including chemo-, radio- or immunotherapy, critically relies on intact cell death programs in cancer cells. Therefore, defects in apoptosis pathways are frequently associated with the resistance to anticancer therapies. In recent years, the identification and characterization of the molecules and pathways that are involved in the regulation and execution of cell death in leukaemia and lymphoma cells, for example tumour necrosis factor related apoptosis inducing ligand (TRAIL), 'inhibitor of apoptosis' (IAP) proteins and Bcl-2, have set the ground for the development of novel diagnostic tools and molecular therapeutics targeting apoptosis pathways in haematological malignancies. PMID- 19298594 TI - Mutation analysis of the HAX1 gene in childhood myelodysplastic syndrome. PMID- 19298595 TI - Expression of genes encoding for proteins involved in heparan sulphate and chondroitin sulphate chain synthesis and modification in normal and malignant plasma cells. AB - Syndecan-1 is a proteoglycan that concentrates heparin-binding factors on the surface of multiple myeloma cells, and probably plays a major role in multiple myeloma biology. As heparan sulphate and chondroitin sulphate are the bioactive components of syndecan-1, we analysed the signature of genes encoding 100 proteins involved in synthesis of these chains, i.e. from precursor uptake to post-translational modifications, using Affymetrix microarrays. The expression of enzymes required for heparan sulphate and chondroitin sulphate biosynthesis was shown to increase in parallel with syndecan-1 expression, throughout the differentiation of memory B cells into plasmablasts and normal bone marrow plasma cells. Sixteen genes were significantly different between normal and malignant plasma cells, nine of these genes -EXT2, CHSY3, CSGALNACT1, HS3ST2, HS2ST1, CHST11, CSGALNACT2, HPSE, SULF2 - encode proteins involved in glycosaminoglycan chain synthesis or modifications. Kaplan-Meier analysis was performed in two independent series of patients: B4GALT7, CSGALNACT1, HS2ST1 were associated with a good prognosis whereas EXT1 was linked to a bad prognosis. This study provides an overall picture of the major genes encoding for proteins involved in heparan sulphate and chondroitin sulphate synthesis and modifications that can be implicated in normal and malignant plasma cells. PMID- 19298596 TI - Mitochondrial ATP-synthase in the entorhinal cortex is a target of oxidative stress at stages I/II of Alzheimer's disease pathology. AB - Oxidative stress has been implicated in the pathogenesis of several neurodegenerative diseases including Alzheimer's disease (AD). Several proteins have been identified as targets of oxidative damage in AD dementia (usually stages V/VI of Braak) and in subjects with mild cognitive impairment associated with middle stages of AD pathology (stage IV of Braak). In this study, we investigate whether brain proteins are locally modified by oxidative stress at the first stages of AD-related pathology when morphological lesions are restricted to the entorhinal and transentorhinal cortices of neurofibrillary pathology (stages I/II of Braak). Using a proteomic approach, we show that the alpha subunit of the mitochondrial adenosine triphosphate (ATP)-synthase is distinctly lipoxidized in the entorhinal cortex at Braak stages I/II compared with age-matched controls. In addition, ATP-synthase activity is significantly lower in Braak stages I/II than age-matched control, while electron transport chain, expressed by the mitochondrial complex I activity, remains not affected. This is the first study showing oxidative damage in the first stage, and clinically silent period, of AD-related pathology characterized by entorhinal and transentorhinal tauopathy. PMID- 19298598 TI - Lesion profiling at primary isolation in RIII mice is insufficient in distinguishing BSE from classical scrapie. AB - Primary isolation of bovine spongiform encephalopathy (BSE) in RIII mice generates a lesion profile believed to be reproducible and distinct from that produced by classical scrapie. This profile, which is characterized by peaks at gray matter areas 1, 4 and 7 (dorsal medulla, hypothalamus and septal nuclei), is used to diagnose BSE on primary isolation. The aim of this study was to investigate whether the BSE agent could be present in sheep diagnosed with classical scrapie, using lesion profiles in RIII mice as a discriminatory method. Sixty-two positive scrapie field cases were collected from individual farms between 1996 and 1999 and bioassayed in RIII mice. Fifty-five of these isolates transmitted successfully to at least one mouse. Of the 31 that produced adequate data to allow lesion profile analysis, 10 showed a consistent profile with peaks at brain areas 1, 4 and 7. All inocula for this subgroup were derived from sheep of genotype ARQ/ARQ. While the 1-4-7-scrapie profile exhibited similarities to BSE in RIII mice at primary isolation, it was distinguishable based on histopathology, immunohistochemistry and cluster analysis. We conclude that caution should be taken to distinguish this profile from BSE and that additional parameters should be considered to reach a final diagnosis. PMID- 19298597 TI - Morpho-physiologic characteristics of dorsal subicular network in mice after pilocarpine-induced status epilepticus. AB - The goal of this study was to examine the morpho-physiologic changes in the dorsal subiculum network in the mouse model of temporal lobe epilepsy using extracellular recording, juxtacellular and immunofluorescence double labeling, and anterograde tracing methods. A significant loss of total dorsal subicular neurons, particularly calbindin, parvalbumin (PV) and immunopositive interneurons, was found at 2 months after pilocarpine-induced status epilepticus (SE). However, the sprouting of axons from lateral entorhinal cortex (LEnt) was observed to contact with surviving subicular neurons. These neurons had two predominant discharge patterns: bursting and fast irregular discharges. The bursting neurons were mainly pyramidal cells, and their dendritic spine density and bursting discharge rates were increased significantly in SE mice compared with the control group. Fast irregular discharge neurons were PV-immunopositive interneurons and had less dendritic spines in SE mice when compared with the control mice. When LEnt was stimulated, bursting and fast irregular discharge neurons had much shorter latency and stronger excitatory response in SE mice compared with the control group. Our results illustrate that morpho-physiologic changes in the dorsal subiculum could be part of a multilevel pathologic network that occurs simultaneously in many brain areas to contribute to the generation of epileptiform activity. PMID- 19298599 TI - Potent antitumor activity of double-regulated oncolytic adenovirus-mediated ST13 for colorectal cancer. AB - Following targeted gene virotherapy, the suppression of tumorigenicity 13 (ST13) gene was inserted into the double-regulated oncolytic adenovirus SG500 to ensure more safety and potent antitumor activity against colorectal cancer in vitro and in vivo. We generated the ST13-expressing oncolytic adenovirus SG500-ST13, with which colorectal carcinoma cell lines SW620 and HCT116, and the lung fibroblast cell line WI38, were infected. Crystal violet staining was carried out to detect the cytopathic effect in cells, and the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide method was used to assay cell viability. The effect of apoptosis induced by SG500-ST13 was confirmed by Hoechst staining and the TdT mediated dUTP-biotin nick-end labeling method. To further identify the antitumor effects of SG500-ST13 on HCT116 xenografts in Balb/c nude mice, the induction of cell death was assessed by hematoxylin-eosin staining. Immunohistochemical study was also carried out. PMID- 19298600 TI - Up-regulated expression of ADAM17 in gastrointestinal stromal tumors: coexpression with EGFR and EGFR ligands. AB - Metalloproteinase activities of a disintegrin and metalloproteinases (ADAMs), matrix metalloproteinases (MMPs), and membrane type (MT-)MMPs are involved in many aspects of tumor biology. ADAMs are transmembrane proteins that cleave membrane-anchored proteins to release soluble factors, and thereby mediate important biological phenomena in tumors. The aim of this study was to analyze histopathology, expression and roles of metalloproteinases, especially ADAMs, in gastric gastrointestinal stromal tumor (GIST). Histopathology and immunohistochemical expression of ADAMs were examined in 89 gastric GISTs. In 11 GISTs, ADAM expression was examined at mRNA and protein levels by reverse transcription-polymerase chain reaction (RT-PCR) and immunoblotting, respectively. RT-PCR analysis showed frequent expression of ADAM9 (91%), ADAM10 (64%), ADAM17 (82%), MMP-2 (82%), and MT1-MMP (73%). However, ADAM17 and MMP-2 were the only metalloproteinases that were up-regulated in GISTs at the protein level compared with non-neoplastic gastric tissues. ADAM17 was immunohistochemically expressed in 93% of GIST versus 16% of normal gastric tissues. Furthermore, CD117-positive interstitial cells of Cajal in normal gastric tissues were all negative for ADAM17 with double immunostaining. Expressions of epidermal growth factor receptor (EGFR) and several EGFR ligands such as amphiregulin, heparin-binding epidermal growth factor (HB-EGF), betacellulin, and epiregulin were also demonstrated in GIST by RT-PCR. Protein expression of EGFR, phosphorylated EGFR, amphiregulin, and HB-EGF, both of which can be shed by ADAM17, was confirmed in tumors coexpressing ADAM17 by immunoblotting. Moreover, proteolytically cleaved soluble forms of amphiregulin were identified in tumor extracts. Considered together, the results suggest that ADAM17 may contribute to the progression and growth of GIST through shedding of EGFR ligands and consequent EGFR stimulation. ADAM17, as a major sheddase in GIST, could be potentially a suitable target in anticancer treatment of imatinib resistant GISTs. PMID- 19298601 TI - Overexpression of EIF-5A2 predicts tumor recurrence and progression in pTa/pT1 urothelial carcinoma of the bladder. AB - The authors investigated the status of abnormalities of eIF-5A2 gene in superficial (pTa/pT1) urothelial carcinoma of the bladder (UC), as well as its correlation with clinicopathologic variables and patient outcome. The methods of immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were utilized to examine protein/mRNA(messenger RNA) expression and amplification of eIF-5A2 in a cohort of pTa/pT1 UCs. Overexpression of EIF-5A2 was examined by IHC in 38/112 (33.9%) pTa/pT1 UCs. A significant association of overexpression of EIF 5A2 with shortened UC patient recurrence-free survival (P = 0.002), as well as with shortened progression-free survival (P = 0.004), was demonstrated. Importantly, multivariate Cox regression analysis revealed that EIF-5A2 expression provided a significant independent prognostic parameter either in tumor recurrence (P = 0.002) or in tumor progression (P = 0.007). FISH results demonstrated that eIF-5A2 amplification was detected in 5/59 of the informative UCs; in each of the five cases with eIF-5A2 amplification, overexpression of EIF 5A2 was observed. In the remaining 54 UCs without eIF-5A2 amplification, 16 cases were also observed to have overexpression of EIF-5A2. In 13 pairs of UC and adjacent normal tissues, eight UCs were examined and showed up-regulated eIF-5A2 mRNA by RT-PCR, while increased expression of EIF-5A2 protein was only detected in 4/8 UCs by Western blotting. These findings suggest that overexpression of EIF 5A2, as detected by IHC, may predict tumor recurrence and progression in pTa/pT1 UC patients, and the protein expression of eIF-5A2 might be regulated not only by gene amplification, but also by other molecular mechanisms. PMID- 19298602 TI - Isoflavone, polymorphisms in estrogen receptor genes and breast cancer risk in case-control studies in Japanese, Japanese Brazilians and non-Japanese Brazilians. AB - Epidemiologic studies have shown an inverse association between isoflavones and breast cancer risk. Because isoflavones bind estrogen receptors, we hypothesized that polymorphisms in the estrogen receptor genes might modify the association between isoflavone intake and breast cancer risk. We conducted hospital-based case-control studies of patients aged 20-74 years with primary, incident, histologically confirmed invasive breast cancer, and matched controls from among medical checkup examinees in Nagano, Japan, and from cancer-free patients in Sao Paulo, Brazil. A total of 846 pairs (388 Japanese, 79 Japanese Brazilians and 379 non-Japanese Brazilians) completed validated food frequency questionnaires, and provided blood samples. Five single nucleotide polymorphisms in the estrogen receptor alpha (rs9340799, rs1913474, and rs2234693) and beta (rs4986938 and rs1256049) genes were genotyped. We found no consistent association between the five single nucleotide polymorphisms and breast cancer risk among the three populations. In analyses of combinations of isoflavone intake and single nucleotide polymorphisms, an inverse association between intake and risk was limited to women with the GG genotype of the rs4986938 polymorphism for postmenopausal Japanese (odds ratio for highest versus lowest tertile = 0.47; P for trend = 0.01), Japanese Brazilians (odds ratio for highest versus lowest median = 0.31) and non-Japanese Brazilians (odds ratio for consumers versus non consumers = 0.37) (P for interaction = 0.11, 0.08, and 0.21, respectively). We found no remarkable difference for the other four polymorphisms. Our findings suggest that polymorphisms in the estrogen receptor beta gene may modify the association between isoflavone intake and breast cancer risk. PMID- 19298603 TI - Stanniocalcin 2 overexpression in castration-resistant prostate cancer and aggressive prostate cancer. AB - Prostate cancer is usually androgen-dependent and responds well to androgen ablation therapy based on castration. However, at a certain stage some prostate cancers eventually acquire a castration-resistant phenotype where they progress aggressively and show very poor response to any anticancer therapies. To characterize the molecular features of these clinical castration-resistant prostate cancers, we previously analyzed gene expression profiles by genome-wide cDNA microarrays combined with microdissection and found dozens of trans activated genes in clinical castration-resistant prostate cancers. Among them, we report the identification of a new biomarker, stanniocalcin 2, as an overexpressed gene in castration-resistant prostate cancer cells. Real-time polymerase chain reaction and immunohistochemical analysis confirmed overexpression of stanniocalcin 2, a 302-amino-acid glycoprotein hormone, specifically in castration-resistant prostate cancer cells and aggressive castration-naive prostate cancers with high Gleason scores (8-10). The gene was not expressed in normal prostate, nor in most indolent castration-naive prostate cancers. Knockdown of stanniocalcin 2 expression by short interfering RNA in a prostate cancer cell line resulted in drastic attenuation of prostate cancer cell growth. Concordantly, stanniocalcin 2 overexpression in a prostate cancer cell line promoted prostate cancer cell growth, indicating its oncogenic property. These findings suggest that stanniocalcin 2 could be involved in aggressive phenotyping of prostate cancers, including castration-resistant prostate cancers, and that it should be a potential molecular target for development of new therapeutics and a diagnostic biomarker for aggressive prostate cancers. PMID- 19298604 TI - HAb18G/CD147 inhibits starvation-induced autophagy in human hepatoma cell SMMC7721 with an involvement of Beclin 1 down-regulation. AB - HAb18G/CD147, a transmembrane glycoprotein highly expressed in various types of malignant cells, mainly functions as an inducer of matrix metalloproteinases to promote tumor growth, invasion and metastasis. However, whether there are other mechanisms underlying the role of HAb18G/CD147 in tumor progression remains to be elucidated. In this study, we investigated the functional effects of HAb18G/CD147 on autophagy in hepatoma cell line SMMC7721 using immunofluorescence staining, Western blot and transmission electronmicroscopy. Our data showed that specific small interference RNA (siRNA) considerably down-regulated the expression of HAb18G/CD147 in SMMC7721 cells at both messenger RNA (mRNA) and protein levels. The down-regulation of HAb18G/CD147 significantly promoted starvation-induced autophagy in a dose-dependent manner. Using trypan blue exclusion assay, we found that HAb18G/CD147 notably enhanced the survival of SMMC7721 cells through inhibiting starvation-induced autophagy. In addition, we demonstrated that HAb18G/CD147 down-regulated the expression of autophagy-regulating protein Beclin 1 in SMMC7721 cells. Furthermore, our data indicated that HAb18G siRNA transfected SMMC7721 cells had a significantly decreased level of phosphorylated serine/threonine protein kinase B (pAkt) and the expression of Beclin 1 was inversely associated with the level of pAkt, suggesting that the Class I phosphatidylinositol 3 kinase-Akt pathway may be involved in the down-regulation of Beclin 1 by HAb18G/CD147. Overall, we provide the first experimental evidence to show that HAb18G/CD147 may play an important role in the inhibitory regulation of autophagy. Therefore, our data suggest a new molecular mechanism for HAb18G mediated hepatoma progression. PMID- 19298605 TI - Cellular distributions of molecules with altered expression specific to thyroid proliferative lesions developing in a rat thyroid carcinogenesis model. AB - To identify differentially regulated molecules related to early and late stages of tumor promotion in a rat two-stage thyroid carcinogenesis model by an antithyroid agent, sulfadimethoxine, microarray-based microdissected lesion specific gene expression profiling was carried out. Proliferative lesions for profiling were divided into two categories: (i) focal follicular cell hyperplasias (FFCH) and adenomas (Ad) as early lesions; and (ii) carcinomas (Ca) as more advanced. In both cases, gene expression was compared with that in surrounding non-tumor follicular cells. Characteristically, upregulation of cell cycle-related genes in FFCH + Ad, downregulation of genes related to tumor suppression and transcription inhibitors of inhibitor of DNA binding (Id) family proteins in Ca, and upregulation of genes related to cell proliferation and tumor progression in common in FFCH + Ad and Ca, were detected. The immunohistochemical distributions of molecules included in the altered expression profiles were further examined. In parallel with microarray data, increased localization of ceruloplasmin, cyclin B1, and cell division cycle 2 homolog A, and decreased localization of poliovirus receptor-related 3 and Id3 were observed in all types of lesion. Although inconsistent with the microarray data, thyroglobulin immunoreactivity appeared to reduce in Ca. The results thus suggest cell cycling facilitation by induction of M-phase-promoting factor consisting of cyclin B1 and cell division cycle 2 homolog A and generation of oxidative responses as evidenced by ceruloplasmin accumulation from an early stage, as well as suppression of cell adhesion involving poliovirus receptor-related 3 and inhibition of cellular differentiation regulated by Id3. Decrease of thyroglobulin in Ca may reflect dedifferentiation with progression. PMID- 19298606 TI - Increased prevalence of autoimmunity in Turner syndrome--influence of age. AB - Individuals with Turner syndrome (TS) are prone to develop autoimmune conditions such as coeliac disease (CD), thyroiditis and type 1 diabetes (T1DM). The objective of the present study was to examine TS of various karyotypes for autoantibodies and corresponding diseases. This was investigated in a prospective cross-sectional study of Danish TS patients (n = 107, median age 36.7 years, range: 6-60 years). A medical history was recorded and a blood sample was analysed for autoantibodies against gliadin, transglutaminase, adrenal cortex, intrinsic factor, anti-thyroid peroxidase (anti-TPO) and glutamic-acid decarboxylase 65 (GAD-65). Autoantibodies were present in 58% (n = 61) of all patients, whereof 18% (11) had autoantibodies targeting more than one organ. Patients with autoantibodies were significantly older than those without (P = 0.001). Anti-TPO was present in 45% (48) of patients, of whom 33% (16) were hypothyroid. Overall, 18% (19) presented with CD autoantibodies, of whom 26% (five) had CD. Anti-TPO and CD autoantibodies co-existed in 9% (10). Immunoglobulin A deficiency was found in 3% (three) of patients, who all had CD autoantibodies without disease. Among four patients with anti-GAD-65 none had T1DM, but two were classified as having T2DM. One patient had adrenocortical autoantibodies but not adrenal failure. Autoantibodies against intrinsic factor were absent. Anti-GAD-65 was increased in isochromosomal karyotypes (3/23 versus 1/84, P = 0.008) with no other association found between autoantibodies and karyotype. In conclusion, TS girls and women face a high prevalence of autoimmunity and associated disease with a preponderance towards hypothyroidism and CD. Thus, health care providers dealing with this patient group should be observant and test liberally for these conditions even before clinical symptoms emerge. PMID- 19298607 TI - Influence of Enterococcus faecium SF68 probiotic on giardiasis in dogs. AB - BACKGROUND: Giardiasis is a common, potentially zoonotic disease, and dogs often harbor and shed cysts without showing clinical signs. Treatment with the probiotic Enterococcus faecium SF68 has been shown to stimulate mucosal and systemic immunity in a variety of animal models and in young dogs, and to reduce giardial cyst and antigen shedding in rodents. HYPOTHESIS: Adult dogs with chronic naturally acquired giardiasis will have decreased giardial fecal cyst and antigen shedding and increased innate and adaptive immunity after 6 weeks probiotic treatment with E. faecium SF68. ANIMALS: Twenty adult dogs. METHODS: After a 6-week dietary equilibration period, dogs were randomized to receive E. faecium SF68 or placebo for 6 weeks, and then crossed over to the alternate treatment. We measured cyst shedding, fecal giardial antigen, fecal immunoglobulin A (IgA) concentration, and circulating leukocyte phagocytic activity at multiple timepoints to determine the effect of E. faecium SF68 on giardiasis and immune responses in these dogs. RESULTS: No differences were observed between placebo or E. faecium SF68 treatment for giardial cyst shedding, fecal antigen shedding, fecal IgA concentration, or leukocyte phagocytic activity. CONCLUSIONS: Short-term treatment with E. faecium SF68 of dogs with chronic naturally acquired subclinical giardiasis fails to affect giardial cyst shedding or antigen content and does not alter innate or adaptive immune responses. PMID- 19298608 TI - Serum acute phase protein concentrations in dogs with autoimmune hemolytic anemia. AB - BACKGROUND: Serial monitoring of acute phase protein (APP) concentrations in canine autoimmune hemolytic anemia (AIHA) has not been reported. HYPOTHESES: Acute canine AIHA is accompanied by an acute phase response (APR) characterized by increased C-reactive protein (CRP) and alpha1-acid glycoprotein (AAG) concentrations and decreased albumin concentrations. ANIMALS: Twenty-seven dogs with AIHA and 11 control dogs. METHODS: Prospective, cohort study. CRP, AAG, and albumin concentrations, white blood cell (WBC) count, and packed cell volume (PCV) were determined at admission (day 1), every 48 hours until death or discharge, and on days 30, 90, 180, and 365. RESULTS: Compared with controls, CRP and AAG concentrations were increased and albumin concentration was decreased in dogs with AIHA (days 1-7; P < .002) and normalized with disease stabilization (days 9-365; P > .05). APP concentrations on day 1 were not predictive of survival, duration of hospitalization, or number of blood transfusions (P= .153 .940). PCV correlated with APP concentrations over time (CRP r=-.600, AAG r= .665, albumin r= .533; P < .0001) as did WBC count (CRP r= .253, AAG r= .486, albumin r=-.246; P < .006). Day 1 CRP concentration was lower for dogs that received corticosteroids before referral (115.3 microg/mL) compared with dogs that did not (191.2 microg/mL; P= .02). CONCLUSIONS: An APR occurs in canine AIHA. Initial APP concentrations are not predictive of acute survival, correlate with hematologic markers of remission, and normalize rapidly with disease stabilization. PMID- 19298609 TI - A rapid detection method for the ryanodine receptor 1 (C7360G) mutation in Quarter Horses. AB - BACKGROUND: Anesthetic-induced malignant hyperthermia has been documented in Quarter Horses and is caused by a single-point mutation in the ryanodine receptor 1 gene at nucleotide C7360G generating a R2454G amino acid substitution. An accurate, faster molecular test that is less prone to contamination would facilitate screening for the mutation in horses intended for breeding, in those undergoing surgical procedures, and in those with clinical signs compatible with malignant hyperthermia. OBJECTIVE: To report a rapid and accurate method for the detection of the ryanodine receptor 1 C7360G mutation. ANIMALS: Eleven diseased, 10 healthy, and 225 randomly selected Quarter Horses. METHODS: This study included horses with the ryanodine receptor 1 C7360G mutation as detected by gene sequencing. Available genomic and complementary DNA extracted from whole blood, hair or skeletal muscle was used for genetic analysis. Real-time polymerase chain reaction (RT-PCR) melting curve analysis was performed by equine specific primers and 2 hybridization probes (sensor and anchor probes) that contain the site of the mutation. Results from this method were blinded and compared with nucleic acid sequencing for validation. RESULTS: A rapid genotyping assay with fluorescence resonance energy transfer probes and melting curve analysis was accurate (100% agreement, K= 1) for identification of affected horses. The prevalence of the mutation in a random population of Quarter Horses was 1.3%. CONCLUSIONS AND CLINICAL IMPORTANCE: Malignant hyperthermia in Quarter Horses can be rapidly and accurately detected by RT-PCR melting curve genotyping with hybridization probes. PMID- 19298610 TI - Ionized hypocalcemia in critically ill dogs. AB - BACKGROUND: Ionized hypocalcemia (iHCa) is a common electrolyte disturbance in critically ill people, especially those with sepsis. The cause of the iHCa is not entirely understood and is likely multifactorial. Critically ill people with iHCa have longer hospital stays and higher mortality rates compared to people with normocalcemia. There are no published clinical studies evaluating the incidence and impact of iHCa in critically ill dogs. HYPOTHESIS: iHCa occurs in critically ill dogs, is more prevalent in dogs with systemic inflammatory response syndrome (SIRS) or sepsis, and is associated with longer hospital stays and higher mortality. ANIMALS: One hundred and forty-one client-owned dogs admitted to a companion animal intensive care unit (ICU) in a veterinary teaching hospital. METHODS: Prospective observational study of sequentially enrolled dogs. Blood was collected and analyzed within an hour of admission from all dogs presented to the ICU that met study inclusion criteria. RESULTS: The incidence of iHCa (iCa < 1.11 mmol/L) was 16%. The presence of iHCa was associated with longer ICU (P= .038) and hospital (P= .012) stays but not with decreased survival (P= .60). Dogs with sepsis as defined by >or=3 SIRS criteria and a positive culture were more likely to have iHCa (P= .050). CONCLUSIONS AND CLINICAL RELEVANCE: In dogs not previously treated with fluids or blood products intravenously, the finding of iHCa upon admission to the ICU predicted a longer duration of ICU and hospital stay. Septic dogs with positive cultures were more likely to have iHCa. PMID- 19298611 TI - Tolerability of gemcitabine and carboplatin doublet therapy in cats with carcinomas. AB - BACKGROUND: This study was performed to determine the toxicity of gemcitabine carboplatin doublet therapy in cats with carcinomas. HYPOTHESIS: Gemcitabine and carboplatin are safe in tumor-bearing cats. ANIMALS: Twenty cats with spontaneously occurring carcinomas. METHODS: A cohort of 6 cats received gemcitabine (2 mg/kg IV) on days 1, 8, and 15 and carboplatin (10 mg/kg IV) immediately after gemcitabine on day 1 of a 21-day cycle. A 2nd cohort of 14 cats received carboplatin 4 hours after gemcitabine on day 1 and gemcitabine on day 8 but not day 15. The cycles were repeated every 21 days. RESULTS: Cats in the 1st cohort received a median of 3.75 cycles per animal (range, 1-6). Two cats (33.3%) developed grade 3 or 4 neutropenia, 1 (16.7%) grade 4 thrombocytopenia, and 1 (16.7%) grade 3 gastrointestinal toxicity. Gemcitabine dose reductions and treatment delays occurred in 1 and 4 cats, respectively. Cats in the 2nd cohort received a median of 2 cycles per animal (range, 0.5-10). Two cats (14.3%) had grade 3 or 4 neutropenia and 1 (7.1%) had grade 3 and 4 gastrointestinal toxicity. One cat required gemcitabine dose reduction and 6 had treatment delays. In the 2nd cohort, of 11 cats with measurable tumors, there was 1 complete response (pancreatic carcinoma) and 1 partial response (squamous cell carcinoma, receiving concurrent nonsteroidal anti-inflammatory drugs). CONCLUSIONS AND CLINICAL IMPORTANCE: Gemcitabine-carboplatin combination appears moderately well tolerated in tumor-bearing cats. Minimal patient benefit suggests that alternative schedules or combinations of gemcitabine with other agents should be explored. PMID- 19298612 TI - Association between waste management and cancer in companion animals. AB - BACKGROUND: Increased cancer rates have been documented in people residing in areas around Naples characterized by illegal dumping and incineration of waste. HYPOTHESIS: Risk of cancer in dogs and cats is associated with waste management. ANIMALS: Four hundred and fifty-three dogs and cats with cancer and 1,554 cancer free animals. METHODS: Hospital-based case-control study in Naples (low danger) and nearby cities having a history of illegal waste dumping (high danger). Odds ratio (OR) between high- and low-danger areas was calculated for all tumors and various malignancies in dogs and cats. RESULTS: An increased risk for cancer development was identified in dogs but not in cats residing in high-danger areas (OR: 1.55; 95% confidence interval: 1.18-2.03; P < .01). A 2.39-fold increased risk of lymphoma (P < .01) accounted for the greater tumor frequency in dogs residing in high-danger areas. The risk of mast cell tumor and mammary cancer did not differ in dogs residing in high- or low-danger areas. CONCLUSIONS AND CLINICAL IMPORTANCE: Waste emission from illegal dumping sites increases cancer risk in dogs residing in high-danger areas. An increased prevalence of lymphoma has been previously recognized in humans living close to illegal waste dumps. Thus, epidemiological studies of spontaneous tumors in dogs might suggest a role for environmental factors in canine and human carcinogenesis and can predict health hazards for humans. PMID- 19298614 TI - Family legacies: short- and long-term fitness consequences of early-life conditions in female European rabbits. AB - 1. Environmental conditions during an animal's early life can have profound long term consequences and affect its fitness. In particular, maternal and sibling effects, which can strongly influence the early growth of altricial mammals may be important. Few studies have investigated the influence of such early-life parameters in small mammals, because in these species the early post-natal stage is difficult to monitor under natural conditions. 2. We quantified the effects of litter size (i.e. number of litter siblings), maternal social rank and age and reproductive history of the mother (i.e. whether or not the mother had given birth to a previous litter during this season), and the individual date of birth and social rank on two fitness components of female European rabbits (Oryctolagus cuniculus L.) from a field enclosure population. Analyses were based on data on survival to maturity of 1836 female pups from 10 annual cohorts, and on lifetime reproductive success (LRS; here: the summed up number of offspring surviving to maturity) of 81 adult females from eight annual cohorts. 3. Both fitness components were correlated with the size of the females' original litter and with the age of their mother. Litter size was related to survival to maturity and to LRS in a nonlinear (quadratic) way being highest in females from medium-sized litters. Maternal age also exerted quadratic effects on LRS, which peaked in females born to 2- to 3-year-old mothers. In contrast, survival to maturity increased with increasing age of the mother. 4. Survival to maturity and LRS were decreased in females born later in the breeding season, likely because of the longer time for growth that early born young enjoy before the winter season. In addition, LRS was lower in females which occupied a higher social rank at the onset of their first breeding season. 5. Our results emphasize that factors during early development, in particular parameters of the early social environment, do not only affect juvenile survival but have the potential to exert long-term fitness consequences throughout life. PMID- 19298613 TI - Neurochemical approaches in the laboratory diagnosis of Parkinson and Parkinson dementia syndromes: a review. AB - The diagnosis of Parkinson disease (PD) is rendered on the basis of clinical parameters, whereby laboratory chemical tests or morphological imaging is only called upon to exclude other neurodegenerative diseases. The differentiation between PD and other diseases of the basal ganglia, especially the postsynaptic Parkinson syndromes multisystem atrophy (MSA) and progressive supranuclear palsy (PSP), is of decisive importance, on the one hand, for the response to an appropriate therapy, and on the other hand, for the respective prognosis of the disease. However, particularly at the onset of symptoms, it is difficult to precisely distinguish these diseases from each other, presenting with an akinetic rigid syndrome. It is not yet possible to conduct a neurochemical differentiation of Parkinson syndromes. Therefore, a reliable biomarker is still to be found that might predict the development of Parkinson dementia. Since this situation is currently the subject of various different studies, the following synopsis is intended to provide a brief summary of the investigations addressing the field of the early neurochemical differential diagnosis of Parkinson syndromes and the early diagnosis of Parkinson dementia, from direct alpha-synuclein detection to proteomic approaches. In addition, an overview of the tested biomarkers will be given with regard to their possible introduction as a screening method. PMID- 19298615 TI - Short- and long-term effects of egg size and feeding frequency on offspring quality in the collared flycatcher (Ficedula albicollis). AB - 1. Despite the central importance for life-history theory, egg-size effects on offspring fitness are still considered ambiguous. Most previous studies were only observational and consequently might suffer from uncontrolled correlations between egg size and parental/territory quality. Even after cross-fostering is performed, direct genetic effects and parental adjustment of post-natal care might confound our estimates of egg-size effects per se. 2. I performed a full cross-fostering experiment in the collared flycatcher (Ficedula albicollis) exchanging the whole clutches between pairs of nests. I statistically controlled for direct genetic effects and parental feeding frequencies. I followed young until recruitment to estimate the long-term effects of egg size and parental provisioning. In addition, I compared the effects obtained in the cross-fostering experiment with those obtained from a set of unmanipulated nests. 3. Egg size per se affected offspring morphology in both the short and long term, while having no effect on offspring survival and immunity. Egg-size effects were not confounded by parental post-natal care and direct genetic effects. 4. The number of care givers was an influential predictor of nestling performance. Apart from the variation caused by this factor, feeding frequencies had no consistent effect on offspring performance. 5. Fitness benefits of large eggs may be difficult to establish due to variation of egg-size effects between years and habitats. Feeding frequency may affect offspring state but offspring state may also affect feeding frequency. Varying causality between feeding rate and offspring state may preclude the detection of a positive effect of the former on the latter. PMID- 19298616 TI - Cascading top-down effects of changing oceanic predator abundances. AB - 1. Top-down control can be an important determinant of ecosystem structure and function, but in oceanic ecosystems, where cascading effects of predator depletions, recoveries, and invasions could be significant, such effects had rarely been demonstrated until recently. 2. Here we synthesize the evidence for oceanic top-down control that has emerged over the last decade, focusing on large, high trophic-level predators inhabiting continental shelves, seas, and the open ocean. 3. In these ecosystems, where controlled manipulations are largely infeasible, 'pseudo-experimental' analyses of predator-prey interactions that treat independent predator populations as 'replicates', and temporal or spatial contrasts in predator populations and climate as 'treatments', are increasingly employed to help disentangle predator effects from environmental variation and noise. 4. Substantial reductions in marine mammals, sharks, and piscivorous fishes have led to mesopredator and invertebrate predator increases. Conversely, abundant oceanic predators have suppressed prey abundances. Predation has also inhibited recovery of depleted species, sometimes through predator-prey role reversals. Trophic cascades have been initiated by oceanic predators linking to neritic food webs, but seem inconsistent in the pelagic realm with effects often attenuating at plankton. 5. Top-down control is not uniformly strong in the ocean, and appears contingent on the intensity and nature of perturbations to predator abundances. Predator diversity may dampen cascading effects except where nonselective fisheries deplete entire predator functional groups. In other cases, simultaneous exploitation of predator and prey can inhibit prey responses. Explicit consideration of anthropogenic modifications to oceanic foodwebs should help inform predictions about trophic control. 6. Synthesis and applications. Oceanic top-down control can have important socio-economic, conservation, and management implications as mesopredators and invertebrates assume dominance, and recovery of overexploited predators is impaired. Continued research aimed at integrating across trophic levels is needed to understand and forecast the ecosystem effects of changing oceanic predator abundances, the relative strength of top-down and bottom-up control, and interactions with intensifying anthropogenic stressors such as climate change. PMID- 19298618 TI - Truth hurts. PMID- 19298619 TI - Potential impact of adjusting the threshold of the quantitative D-dimer based on pretest probability of acute pulmonary embolism. AB - OBJECTIVES: The utility of D-dimer testing for suspected pulmonary embolism (PE) can be limited by test specificity. The authors tested if the threshold of the quantitative D-dimer can be varied according to pretest probability (PTP) of PE to increase specificity while maintaining a negative predictive value (NPV) of >99%. METHODS: This was a prospective, observational multicenter study of emergency department (ED) patients in the United States. Eligible patients had a diagnostic study ordered to evaluate possible PE. PTP was determined by the clinician's unstructured estimate and the Wells score. Five different D-dimer assays were used. D-dimer test performance was measured using 1) standard thresholds and 2) variable threshold values: twice (for low PTP patients), equal (intermediate PTP patients), or half (high PTP patients) of standard threshold. Venous thromboembolism (VTE) within 45 days required positive imaging plus decision to treat. RESULTS: The authors enrolled 7,940 patients tested for PE, and clinicians ordered a quantitative D-dimer for 4,357 (55%) patients who had PTPs distributed as follows: low (74%), moderate (21%), or high (4%). At standard cutoffs, across all PTP strata, quantitative D-dimer testing had a test sensitivity of 94% (95% confidence interval [CI] = 91% to 97%), specificity of 58% (95% CI = 56% to 60%), and NPV of 99.5% (95% CI = 99.1% to 99.7%). If variable cutoffs had been used the overall sensitivity would have been 88% (95% CI = 83% to 92%), specificity 75% (95% CI = 74% to 76%), and NPV 99.1% (95% CI = 98.7% to 99.4%). CONCLUSIONS: This large multicenter observational sample demonstrates that emergency medicine clinicians currently order a D-dimer in the majority of patients tested for PE, including a large proportion with intermediate PTP and high PTP. Varying the D-dimer's cutoff according to PTP can increase specificity with no measurable decrease in NPV. PMID- 19298620 TI - Who will consent to emergency treatment trials for subarachnoid hemorrhage? AB - OBJECTIVES: Aneurysmal subarachnoid hemorrhage (SAH) is a devastating disorder that still requires much clinical study. However, the decision to participate in a randomized clinical trial, particularly a neuroemergency trial, is a complex one. The purposes of this survey were to determine who would participate in a randomized clinical trial that intended to examine transfusion practices after SAH, to identify who could serve as potential proxy decision-makers, and to find which patient characteristics were associated with the decision to participate. METHODS: This was a cross-sectional study using a self-administered questionnaire, composed of a brief description of the proposed trial followed by questions about participation using a 5-point Likert scale. Information sought included potential decision-maker, demographic data, setting and reason for current health care access, and personal or family history of neurologic injury. RESULTS: Nine-hundred five subjects were enrolled during emergency department (ED) visits, office visits, hospital admissions, or online, during a 1-month period: 63% were women and 46% were white. Nonneurologic problems were the leading reason (90%) for health care access, but 45% had a personal or family history of neurologic injury. Overall, 54% (95% confidence interval [CI] = 51% to 57%) of subjects stated they would definitely or probably consent to participate. No subject characteristics were associated with this decision: age (p = 0.28), sex (p = 0.16), race/ethnicity (p = 0.07), education (p = 0.44), religion (p = 0.42), clinical setting (p = 0.14), reason for visit (p = 0.58), and/or history of neurologic injury (p = 0.33). The vast majority (88%) identified a family member as the proxy decision-maker, again without differences among groups. CONCLUSIONS: Greater than half of respondents stated they would participate in a proposed emergency treatment trial for SAH. Our survey suggests that the decision to participate is highly individualized, because no demographic, pathologic, historical, or access-related predictors of choice were found. Educational materials designed for this type of trial would need to be broad-based. Family members should be considered as proxy decision-makers where permitted by federal and local regulations. PMID- 19298621 TI - Predictors of hospital admission for chronic obstructive pulmonary disease exacerbations in Canadian emergency departments. AB - OBJECTIVES: The objective was to examine predictors of hospital admission among adults presenting to Canadian emergency departments (EDs) for acute exacerbations of chronic obstructive pulmonary disease (COPD). Current acute treatment approaches and outcomes 2 weeks after the ED visit are also described. METHODS: Subjects, aged > or =35 years presenting with COPD exacerbations to 16 EDs across Canada, underwent a structured in-ED interview and a telephone interview 2 weeks later. RESULTS: Of 501 study patients, 247 (49.3%; 95% confidence interval [CI] = 44.9% to 53.6%) were admitted. Admitted patients were older, were more often former smokers, and had more admissions for COPD during the past 2 years. They also reported more days of activity limitation and use of inhaled beta(2) agonists in the previous 24 hours. Canadian Triage and Acuity Scale (CTAS), respiratory rate (RR), and airflow obstruction were more severe in the hospitalized group. Most of the patients received inhaled beta(2)-agonists, anticholinergics, oral corticosteroids (CS), and antibiotics; hospitalized patients received more aggressive treatments. The median ED length of stay (LOS) of admitted patients was 13.1 hours (interquartile range [IQR] = 7.4-23.0) compared to 5.6 hours (IQR = 4.2-8.4) in discharged patients. Admission was associated with at least two COPD admissions in the past 2 years (odds ratio [OR] = 2.10; 95% CI = 1.24 to 3.56), receiving oral CS for COPD (OR = 1.72; 95% CI = 1.08 to 2.74), having a CTAS score of 1-2 (OR = 2.04; 95% CI = 1.33 to 3.12), and receiving adjunct ED treatments (OR = 3.95; 95% CI = 2.45 to 6.35). Use of EDs for usual COPD care was associated with a reduced risk of admission (OR = 0.43; 95% CI = 0.28 to 0.66). CONCLUSIONS: Exacerbations of COPD in Canadian EDs result in prolonged ED stays and approximately 50% hospitalization despite aggressive acute treatment approaches. Historical, severity, and treatment-related factors were strongly associated with hospital admission. Validation of these results should be completed prior to widespread use. PMID- 19298622 TI - Maternal cardiovascular disease risk in relation to the number of offspring born small for gestational age: national, multi-generational study of 2.7 million births. AB - AIM: To investigate the risk of small for gestational age (SGA) births in relation to maternal history of cardiovascular disease (CVD) across two generations and additionally to analyse maternal CVD risk based on number of SGA offspring. METHODS: We used register data from 1.4 million women and 2.7 million offspring. The outcome measures were risk of being SGA in relation to maternal total CVD (n = 10 436) across two generations, as well as risk of CVD in mothers in relation to the number of their SGA offspring, stratified by educational level. RESULTS: Compared to no family history of CVD (reference) the hazard ratio (HR) for being SGA in female offspring was 1.11 (95% confidence interval (CI) 1.09-1.13) for a positive maternal history of CVD. The highest risk was shown in daughters when both the mother and the grandmother had a history of CVD (HR 1.32, 95% CI 1.24-1.39). There was a stepwise increased risk of CVD events in mothers in relation to the number of their SGA offspring (HR 1.41-1.86) when 'no SGA offspring' was used as reference. The risk of CVD in relation to SGA status was increased in the least educated group (HR 2.7-5.0) compared to the group with the highest level of education with no SGA offspring. CONCLUSION: The risk of SGA offspring and the risk of maternal CVD are mutually interdependent and both conditions increased in women with a low level of education. PMID- 19298623 TI - Long hospitalization is the most important risk factor for early weaning from breast milk in premature babies. AB - AIM: To identify certain variables related to the infants' course that might have an impact on the mothers' decision to breastfeed. PATIENTS AND METHOD: Retrospective survey including all patients <1500 g birth weight (BW) treated between January 1, 2000 and December 31, 2005 at the Neonatal Intensive Care Unit of the Medical University of Vienna who were not transferred to another hospital. Multiple regression analysis of the following variables was carried out: gestational age (GA), BW, length of stay (LOS), parity, singleton or multiple gestation, sex and severe morbidity. RESULTS: Of the 239 patients included, 142 (60%) were fed breast milk at the time of final discharge, 97 (40%) were fed formula. LOS was significantly correlated with the probability of being breastfed: the shorter it was, the higher was the probability of being breastfed at the time of final discharge (p = 0.0064 for singletons, p = 0.001 for multiples). Lower GA also increased the probability of being breastfed, but this was only statistically significant for multiples (p = 0.001). CONCLUSION: This study shows clearly that the most important influencing factor on the mothers' decision to continue breastfeeding is the LOS. Thus more emphasis should be put on encouraging mothers to continue lactation throughout their babies' hospital stay. PMID- 19298624 TI - The cognitive control of distraction by novelty in children aged 7-8 and adults. AB - The study focused on the development of cognitive control of distraction. Novel sounds were interspersed in a sequence of a constant environmental sound, while subjects were engaged in a task not related to novelty. In both children (7-8 years) and adults, unpredictable novel sounds caused prolonged reaction times (RT), the P3a and the Reorienting Negativity (RON) components of the event related potential, indicating distraction and reorienting of attention. With predictable novels, RT prolongation and RON-amplitude were reduced in both groups, whereas P3a-amplitude reduction was confined to adults. Thus, although children reveal some indication for control of distraction, they do not yet achieve the level of adults. This differential pattern of the development of RT prolongation, P3a, and RON across age groups indicates different maturation of processes involved in the control of distraction and suggests partly independent underlying processes. PMID- 19298625 TI - Association between longitudinal changes in depressive symptoms and plasma fibrinogen levels in school teachers. AB - Depression and anxiety previously predicted coronary artery disease (CAD) risk. Inflammation contributes to CAD and shows an association with depression. We followed 57 teachers (mean 49+/-8 years) over 21 months and investigated whether changes in depressive and anxiety symptoms relate to those in the CAD risk and inflammation marker fibrinogen and vice versa. Increase in depressive symptoms and in fibrinogen levels were significantly correlated. While controlling for baseline depression rendered the association between changes in depression and fibrinogen nonsignificant, taking into account baseline fibrinogen levels maintained the predictive value of fibrinogen change for depression change. Anxiety and fibrinogen changes were not significantly correlated. This dynamic relationship between depression and the inflammatory biomarker fibrinogen might advance our knowledge about psychobiological mechanisms underlying both CAD and sickness behavior. PMID- 19298626 TI - Validation of regression-based myogenic correction techniques for scalp and source-localized EEG. AB - EEG and EEG source-estimation are susceptible to electromyographic artifacts (EMG) generated by the cranial muscles. EMG can mask genuine effects or masquerade as a legitimate effect-even in low frequencies, such as alpha (8-13 Hz). Although regression-based correction has been used previously, only cursory attempts at validation exist, and the utility for source-localized data is unknown. To address this, EEG was recorded from 17 participants while neurogenic and myogenic activity were factorially varied. We assessed the sensitivity and specificity of four regression-based techniques: between-subjects, between subjects using difference-scores, within-subjects condition-wise, and within subject epoch-wise on the scalp and in data modeled using the LORETA algorithm. Although within-subject epoch-wise showed superior performance on the scalp, no technique succeeded in the source-space. Aside from validating the novel epoch wise methods on the scalp, we highlight methods requiring further development. PMID- 19298627 TI - The transferable placebo effect from pain to emotion: changes in behavior and EEG activity. AB - Past studies indicate that the placebo expectation established by analgesic treatment significantly relieves pain perception, while ataractic treatment significantly alleviates unpleasant arousal evoked by negative picture processing. But it is unclear whether the placebo effect can be transferred from one domain to the other, namely from pain to emotion. In this study we led participants to believe in the analgesic effect of magnetic treatment equipment (the placebo) by secretly reducing the intensity of pain stimulus. Then, we examined if this placebo could significantly alter participants' negative affect evoked by watching unpleasant pictures. Our results indicated a significant transferable placebo effect that alleviated negative feelings. EEG recordings showed the transferable placebo treatment induced decreased P2 amplitude and increased N2 amplitude, with source location near the posterior cingulate. PMID- 19298628 TI - Reduced event-related low frequency EEG activity in schizophrenia during an auditory oddball task. AB - This study examines EEG low frequency characteristics which have been linked to specific cognitive functions such as stimulus encoding and attention during an auditory oddball task in schizophrenia patients and healthy controls. EEG data was recorded from 17 young schizophrenia patients in a stable phase of their illness and 17 healthy controls performing an auditory oddball task. Evoked and induced delta and theta activity, N100, P300 amplitude were computed. Between 200 500 ms after a stimulus was presented, patients displayed significantly reduced P300, less evoked and induced delta and theta activity than controls. We conclude that the well known finding of P300 reduction in schizophrenia can be linked to reductions in delta and theta activity, which are a manifestation of impaired stimulus evaluation, memory retrieval, and a lack of sustained attention. PMID- 19298629 TI - Masked repetition and translation priming in second language learners: a window on the time-course of form and meaning activation using erps. AB - Event-related potentials (ERPs) and masked translation priming served to examine the time-course of form and meaning activation during word recognition in second language learners. Targets were repetitions of, translations of, or were unrelated to the immediately preceding prime. In Experiment 1 all targets were in the participants' L2. In Experiment 2 all targets were in the participants' L1. In Exp 1 both within-language repetition and L1-L2 translation priming produced effects on the N250 component and the N400 component. In Experiment 2 only within language repetition produced N250 effects, while both types of priming produced N400 effects. These results suggest rapid involvement of semantic representations during on-going form-level processing of printed words, and an absence of facilitatory connections between the form representations of non-cognate translation equivalents in L2 learners. The implications for bilingual theories of word processing are discussed. PMID- 19298630 TI - Array-based genomics in glioma research. AB - Over the years, several relevant biomarkers with a potential clinical interest have been identified in gliomas using various techniques, such as karyotype, microsatellite analysis, fluorescent in situ hybridization and chromosome comparative genomic hybridization. Despite their pivotal contribution to our understanding of gliomas biology, clinical application of these approaches has been limited by technological and clinical complexities. In contrast, genomic arrays (array-based comparative genomic hybridization and single nucleotide polymorphisms array) have emerged as promising technologies for clinical use in the setting of gliomas. Indeed, their feasibility and reliability have been rigorously assessed in gliomas and are discussed in this review. The well-known genomic biomarkers in gliomas are in fact readily and reliably identified using genomic arrays. Moreover, it detects a multitude of new cryptic genomic markers, with potential biological and/or clinical significances. The main studies dedicated to genomic characterization of gliomas using genomic arrays are reviewed here. Interestingly, several recurrent genomic signatures have been reported by different teams, suggesting the validity of these genomic patterns. In light of this, genomic arrays are relatively simple and cost-effective techniques whose implementation in molecular diagnostic laboratories should be encouraged as a valuable clinical tool for management of glioma patients. PMID- 19298631 TI - Lithium restores neurogenesis in the subventricular zone of the Ts65Dn mouse, a model for Down syndrome. AB - Down syndrome (DS), a high-incidence genetic pathology, involves brain hypoplasia and mental retardation. Emerging evidence suggests that reduced neurogenesis may be a major determinant of brain underdevelopment in DS. To establish whether it is possible to improve neurogenesis in DS, Ts65Dn mice--the most widely used model for DS--and euploid mice were treated with control or lithium chow for 1 month. During the last 3 days animals received one daily injection of 5-bromo-2 deoxyuridine (BrdU)--a marker of proliferating cells--and were sacrificed 24 h after the last injection. Neurogenesis was examined in the subventricular zone (SVZ), a region that retains a neurogenic potential across life. We found that Ts65Dn mice had less (-40%) BrdU+ cells than euploid mice, indicating severe proliferation impairment. Treatment with lithium increased the number of Brdu+ cells in both euploid and Ts65Dn mice. In the latter the number of Brdu+ cells became similar to that of untreated euploid mice. Our study shows that lithium is able to restore cell proliferation in the SVZ of the Ts65Dn mouse and point at treatments with mood stabilizers as a potential tool to improve neurogenesis in patients with DS. PMID- 19298632 TI - The roads ahead: collision risks, trends, and safety of drivers. AB - The propensity of fatal traffic collisions transcends driver age and reinforces the need to evaluate, among other factors, the impact of roadway lighting and other features of driver vision, perception, and performance. Collisions may result from a driver's inability to notice delineation, recognize warnings, and other possible road safety controls during various lighting conditions. Hence we compare the relative accident involvement ratio (RAIR) of collisions of millions of drivers from two U.S. States over an 11-year period, 1991-2001. We associate collision trends through RAIR with bathtub curves that are commonly identified with product failure and reliability engineering. Hence we observe the need for improved and automated driver's license testing techniques and applications, especially as these relate to the visual and cognitive abilities of drivers of all ages. Our findings may ultimately improve motorist safety, save lives, and benefit numerous other states, countries, and agencies, including, but not limited to, aviation, commercial vehicles, maritime, and rail sectors, among others. PMID- 19298633 TI - Erythropoietin receptor is expressed in meningiomas and lower levels are associated with tumour recurrence. AB - AIMS: The Epo-EpoR pathway plays a role in tumour growth, metastasis and treatment resistance and is a potential target in oncological treatment. As the EpoR status in human meningiomas is unknown, our aim was to characterize EpoR expression in these tumours. METHODS: We examined 131 meningioma samples of all WHO grades from 116 patients by immunohistochemistry for EpoR. Among these, 25 meningiomas showed brain invasion and 29 patients had a further tumour recurrence. A group of 20 patients without tumour recurrence served as controls. In 12 cases we were able to compare both the primary and the following recurrent tumours. The presence of EpoR in meningiomas was confirmed by RT-PCR and Western blot. RESULTS: EpoR was expressed in all meningiomas. Statistical analysis revealed that the mean expression levels of EpoR were significantly lower in primary tumours with known recurrence compared with a recurrence-free control group. Additional matched pair analysis in individual cases showed no significant differences between primary and recurrent tumours. No significant correlation between EpoR expression and WHO grade, age, sex or brain invasion was detected. Using specific primer pairs for RT-PCR, we were able to detect all three known isoforms of EpoR: the full-length isoform EpoR-F, the truncated isoform EpoR-T and the soluble isoform EpoR-S. CONCLUSIONS: Our results demonstrate the expression of EpoR in meningiomas. Lower EpoR mean levels might be a useful marker for a higher recurrence risk, but further studies are needed to clarify the influence of EpoR on recurrences and the role of the different isoforms. PMID- 19298634 TI - Recent advances in molecular targeted therapy for metastatic renal cell carcinoma. AB - Advanced renal cell carcinoma (RCC) is resistant to chemotherapy and radiotherapy. Immunotherapy is relatively effective against RCC. However, the response rate is approximately 15-20%. Therefore, new therapeutic approaches are necessary. Recently, molecular mechanisms responsible for the proliferation of RCC are identified, and molecular targeted therapy is developed. Bevacizumab, sorafenib, sunitinib, axitinib, temsirolimus, everolimus are promising molecular targeted therapeutic agents for metastatic RCC, and will be used widely in clinics in the near future. In addition, combination therapy with molecular targeted therapy and other therapies including immunotherapy may also be developed soon. PMID- 19298635 TI - A randomized trial of lower leg and height growth in children with asthma treated with inhaled budesonide from a new dry powder inhaler. AB - Short-term knemometry is a highly sensitive and accurate method for non-invasive assessment of systemic activity of inhaled corticosteroids in children with asthma. However, there are no randomized data available to elucidate the relation between inhaled corticosteroid suppressed short-term lower leg and height growth. The aim of the present study was to assess the relation between short-term lower leg and 1-yr height growth in children with asthma treated with inhaled budesonide from the new Pulairmax inhaler 200 MUg once daily in the morning or montelukast 5 mg once daily. A total of 52 pre-pubertal children with asthma were included in a randomized open-label parallel group study. Length of the lower leg and height were measured by knemometry and stadiometry, respectively, at study entry and after 2, 4, 12, 20, 28, 36, 44 and 52 wks. Lower leg and height growth rates were significantly lower in the budesonide than in the montelukast group (p < 0.0001). Mean 2-wks lower leg growth rate was 0.17 mm/wk in the budesonide and 0.39 mm/wk in the montelukast treated children (p = 0.02). Mean 1-yr height growth rate was 5.51 cm/yr in the budesonide and 6.51 cm/yr in the montelukast group [95% CI: (0.20; 1.79)]. There was a strong linear correlation between lower leg and height growth in both groups, rho = 0.96 (budesonide) and 0.98 (montelukast). In conclusion, 1-yr height growth suppression of budesonide 200 MUg administered via the Pulairmax inhaler once daily in the morning was indicated from suppressed short-term lower leg growth providing evidence that short-term knemometry is able to predict 1-yr height growth suppression of inhaled corticosteroids. Short-term knemometry should be performed as part of the safety assessments of new inhaled corticosteroids and inhalation devices in children with asthma before long-term height growth evaluations are initiated. PMID- 19298636 TI - Hepatitis antibody profile of Royal Thai Army nursing students. AB - We report on a viral hepatitis serosurvey done on 381 nursing students in Bangkok; 360 (94%) were female. The mean age was 20 (+/-3.6) years (range 16-41 years); 143, 92, 86 and 59 students came from Thailand's Central, North, Northeast and South provinces, respectively. The overall prevalence of hepatitis A, B, C and E antibody in the students was 8.9%, 10.8%, 0.5% and 11.5%, respectively. The highest seroprevalence to hepatitis A was observed in cadets from the southern region of Thailand. Seroprevalence rates to hepatitis A and B were less than in other reported studies in Thailand. PMID- 19298637 TI - Increased CXCL-13 levels in human African trypanosomiasis meningo-encephalitis. AB - OBJECTIVES: To determine the role of the B-cell attracting chemokine CXCL-13, which may initiate B-cell trafficking and IgM production in diagnosing HAT meningo-encephalitis. METHODS: We determined CXCL-13 levels by ELISA on paired sera and CSF of 26 patients from Angola and of 16 controls (six endemic and ten non-endemic). Results were compared to standard stage determination markers and IgM intrathecal synthesis. RESULTS: CXCL-13 levels in patients' sera had a median value of 386.6 pg/ml and increased levels were associated with presence of trypanosomes in the CSF but not with other stage markers. CXCL-13 levels in patients' CSF had a median value of 80.9 pg/ml and increased levels were associated with all standard stage determination markers and IgM intrathecal synthesis. CONCLUSION: CXCL-13 levels in CSF increased significantly during the course of HAT. Hence the value of CXCL-13 for diagnosis, follow-up or as a marker of disease severity should be tested in a well-defined cohort study. PMID- 19298638 TI - Judging respiratory specimen acceptability for AFB microscopy: visual vs. microscopic screening. AB - OBJECTIVES: To investigate whether visual assessment or microscopic grading of the cellular composition of specimens is a better strategy to screen specimen quality for tuberculosis (TB) diagnosis. METHODS: About 2643 specimens were collected from TB suspects at the Federal TB centre in Pakistan. Specimens were classified as sputum or saliva visually and microscopically using the criteria proposed by McCarter and Robinson, Van Scoy, Geckler et al., Murray and Washington and Bartlett. The acid-fast bacilli (AFB)-positivity of specimens was also assessed. Results Despite being the least 'technical' and time consuming, visual assessment rejected the lowest proportion of AFB-positive specimens (0.3%). Most microscopic grading criteria, particularly those that considered the squamous epithelial cell count, rejected a large proportion of specimens (30 66%), of which a sizeable fraction contained AFB (6-12%). CONCLUSIONS: Our results indicate that visual assessment by trained technicians is more effective and suitable than microscopic grading for screening specimens for TB diagnosis. TB control resources could be better allocated to optimizing visual screening than investing in more 'strict' microscopic grading tools. PMID- 19298639 TI - Randomized trial to compare the dosage of darbepoetin alfa by administration route in haemodialysis patients. AB - AIM: The doses of darbepoetin alfa required to maintain target haemoglobin levels after s.c. or i.v. administration when recombinant human erythropoietin (rHuEpo) treatment was replaced by darbepoetin alfa treatment in haemodialysis (HD) patients were compared. METHODS: In this prospective, randomized, open-label study, 65 HD patients who were receiving stable SC doses of rHuEpo were switched to an equivalent dose of darbepoetin alfa at a reduced frequency by s.c. or i.v. administration. Patients were randomly assigned to the s.c. group (n = 32) or the i.v. group (n = 33). Darbepoetin alfa doses were titrated to maintain target haemoglobin levels of 8.0-11.0 g/dL for up to 24 weeks. A period of 20 weeks was used for dose titration and haemoglobin stabilization. This was followed by a 4 week evaluation period. RESULTS: The mean haemoglobin concentration during the evaluation period was similar in the s.c. and i.v. groups. The mean dose and mean weight-standardized dose of darbepoetin alfa during the evaluation period tended to be lower in the s.c. group than the i.v. group, although these differences were not statistically significant. The mean weekly darbepoetin alfa dose requirements during the evaluation period significantly decreased in both groups compared to the dose requirements at randomization. CONCLUSION: There is a possibility that s.c. administration of darbepoetin alfa is more efficacious than i.v. administration, but a definite benefit cannot be demonstrated with the current sample size. A bigger sample size is needed to confirm the result. PMID- 19298640 TI - Effect of activin A on tubulointerstitial fibrosis in diabetic nephropathy. AB - AIM: The effect of activin A on tubulointerstitial fibrosis in diabetic nephropathy (DN) using streptozotocin (STZ)-induced diabetic rats and high glucose-cultured HK-2 cells was investigated. METHODS: Male Wistar rats were randomized into a normal control group (NC) and diabetes mellitus group (DM). Diabetes was induced by i.p. injection of STZ. Six rats were respectively killed 4, 8, 12 and 16 weeks after model establishment in each group. The changes of kidney weight/bodyweight (KW/BW), urine albumin excretion rate (AER) and creatinine clearance rate (Ccr) were determined. The morphology of tubulointerstitium was observed by light microscopy. Further biochemical analysis was provided using immunohistochemistry and real-time polymerase chain reaction. The different parameters in high glucose-cultured HK-2 cells were monitored by western blotting or enzyme-linked immunosorbent assay (ELISA) and the intervention of rh-follistatin on them was investigated. RESULTS: Compared with the NC group, there was marked enlargement in the levels of KW/BW, AER, Ccr and interstitial fibrosis index, and the production of P-Smad2/3 and fibronectin in the DM group from 8 to 16 weeks. Activin betaA, mainly located in tubular epithelial cells, was significantly higher in the DM group than that in the NC group throughout the study periods. Follistatin was abundant in the NC group, but was diminished gradually in the DM group. High glucose may facilitate the synthesis of activin betaA, transforming growth factor (TGF)-beta, P-Smad2/3 and fibronectin in HK-2 cells while rh-follistatin inhibited them except TGF-beta. CONCLUSION: Activin A is involved in tubulointerstitial fibrosis in DN by inducing the production of fibronectin through Smad signal pathway. PMID- 19298641 TI - Intravenous calcitriol versus paricalcitol in haemodialysis patients with severe secondary hyperparathyroidism. AB - AIM: Secondary hyperparathyroidism (SHPT) is common among haemodialysis patients. Intensive treatment with calcitriol is often complicated by hypercalcaemia, hyperphosphataemia and elevated calcium phosphorus (Ca X PO(4)) product. Paricalcitol is a vitamin D analogue developed to overcome some of the limitations of calcitriol therapy. The study objectives were to compare the response of intact parathyroid hormone (iPTH) and the incidence of hypercalcaemia, hyperphosphataemia and elevated Ca X PO(4) product in patients with severe SHPT treated with either i.v. calcitriol or i.v. paricalcitol. METHODS: This was a single centre randomized open label study. Patients with serum intact iPTH of 50 pmol/L or more were randomized to receive either i.v. calcitriol (0.01 ug/kg) or i.v. paricalcitol (0.04 ug/kg) during every haemodialysis treatment. Serum iPTH, calcium, phosphorus and alkaline phosphatase were measured at the beginning of the study and every 3 weeks for 12 weeks. RESULTS: Twenty-five patients were enrolled into the study - 12 were randomized into the calcitriol group and 13 into the paricalcitol group. There were no differences in the baseline study parameters between both groups. Serum iPTH levels were significantly reduced (P = 0.003) only in the paricalcitol group but not in the calcitriol group (P = 0.101). On the other hand, serum calcium levels were significantly increased only in the calcitriol group (P = 0.004 vs P = 0.242). Serum phosphorus, alkaline phosphatase and Ca X PO(4) product were not different. CONCLUSION: Intravenous paricalcitol may be superior to i.v. calcitriol for the treatment of severe SHPT in our chronic haemodialysis population. A larger randomized controlled trial is indicated to confirm these initial findings. PMID- 19298642 TI - Histopathological changes and tumour necrosis factor-alpha, transforming growth factor-beta and tenascin expression in patients with primary type I membranoproliferative glomerulonephritis in remission. AB - AIM: Primary type I membranoproliferative glomerulonephritis (MPGN) is a rare cause of glomerular disease with a high relapse rate and poor prognosis. The aim of this study was: (i) to evaluate the histopathological findings associated with remission; and (ii) to document the possible clinical and histopathological factors predicting relapses. METHODS: Eleven type I MPGN patients (five men, six women; mean age, 38.8+/-13.5 years) who were in remission for at least 1 year after the cessation of immunosuppressive drugs were re-biopsied. The intensity of immunostaining for tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, and tenascin was graded from 0 (no staining) to 3+ (maximum staining). RESULTS: Mean baseline mesangial cellularity score and tubulointerstitial infiltration score were reduced and mesangial matrix expansion score was increased at protocol re-biopsies compared to baseline. The glomerular and tubulointerstitial staining scores for TGF-beta1 and tenascin were higher than that of baseline. Reduced tubulointerstitial TNF-alpha expression was found in re-biopsy specimens compared to baseline. Patients have been followed for a mean time of 51.5+/-22.2 months after the protocol biopsy. Eight patients had a relapse. Mesangial cellularity score and glomerular tenascin expression at re biopsy specimens were higher in relapsed patients compared to those without a relapse. CONCLUSION: Our study shows that mesangial cellularity and tubulointerstitial cell infiltration are reducing whereas mesangial matrix expansion, glomerular and tubulointerstitial TGF-beta1 and tenascin expression are increasing with remission. The higher mesangial cell proliferation and glomerular tenascin scores in remission are associated with the development of relapse. PMID- 19298643 TI - Viral and murine interleukin-10 are correctly processed and retain their biological activity when produced in tobacco. AB - BACKGROUND: Interleukin-10 (IL-10) is a potent anti-inflammatory cytokine, with therapeutic applications in several autoimmune and inflammatory diseases. Oral administration of this cytokine alone, or in combination with disease-associated autoantigens could confer protection form the onset of a specific autoimmune disease through the induction of oral tolerance. Transgenic plants are attractive systems for production of therapeutic proteins because of the ability to do large scale-up at low cost, and the low maintenance requirements. They are highly amenable to oral administration and could become effective delivery systems without extensive protein purification. We investigated the ability of tobacco plants to produce high levels of biologically-active viral and murine IL-10. RESULTS: Three different subcellular targeting strategies were assessed in transient expression experiments, and stable transgenic tobacco plants were generated with the constructs that yielded the highest accumulation levels by targeting the recombinant proteins to the endoplasmic reticulum. The best yields using this strategy in T1 plants were 10.8 and 37.0 microg/g fresh leaf weight for viral and murine IL-10, respectively. The recombinant proteins were purified from transgenic leaf material and characterized in terms of their N-glycan composition, dimerization and biological activity in in vitro assays. Both molecules formed stable dimers, were able to activate the IL-10 signaling pathway and to induce specific anti-inflammatory responses in mouse J774 macrophage cells. CONCLUSION: Tobacco plants are able to correctly process viral and murine IL-10 into biologically active dimers, therefore representing a suitable platform for the production for these cytokines. The accumulation levels obtained are high enough to allow delivery of an immunologically relevant dose of IL-10 in a reasonable amount of leaf material, without extensive purification. This study paves the way to performing feeding studies in mouse models of autoimmune diseases, that will allow the evaluation the immunomodulatory properties and effectiveness of the viral IL-10 in inducing oral tolerance compared to the murine protein. PMID- 19298644 TI - Independent evolution of neurotoxin and flagellar genetic loci in proteolytic Clostridium botulinum. AB - BACKGROUND: Proteolytic Clostridium botulinum is the causative agent of botulism, a severe neuroparalytic illness. Given the severity of botulism, surprisingly little is known of the population structure, biology, phylogeny or evolution of C. botulinum. The recent determination of the genome sequence of C. botulinum has allowed comparative genomic indexing using a DNA microarray. RESULTS: Whole genome microarray analysis revealed that 63% of the coding sequences (CDSs) present in reference strain ATCC 3502 were common to all 61 widely-representative strains of proteolytic C. botulinum and the closely related C. sporogenes tested. This indicates a relatively stable genome. There was, however, evidence for recombination and genetic exchange, in particular within the neurotoxin gene and cluster (including transfer of neurotoxin genes to C. sporogenes), and the flagellar glycosylation island (FGI). These two loci appear to have evolved independently from each other, and from the remainder of the genetic complement. A number of strains were atypical; for example, while 10 out of 14 strains that formed type A1 toxin gave almost identical profiles in whole genome, neurotoxin cluster and FGI analyses, the other four strains showed divergent properties. Furthermore, a new neurotoxin sub-type (A5) has been discovered in strains from heroin-associated wound botulism cases. For the first time, differences in glycosylation profiles of the flagella could be linked to differences in the gene content of the FGI. CONCLUSION: Proteolytic C. botulinum has a stable genome backbone containing specific regions of genetic heterogeneity. These include the neurotoxin gene cluster and the FGI, each having evolved independently of each other and the remainder of the genetic complement. Analysis of these genetic components provides a high degree of discrimination of strains of proteolytic C. botulinum, and is suitable for clinical and forensic investigations of botulism outbreaks. PMID- 19298645 TI - Relation between psychosocial variables and the glycemic control of patients with type 2 diabetes: a cross-sectional and prospective study. AB - BACKGROUND: This cross-sectional and prospective study used a variety of psychological inventories to evaluate the relationship between psychosocial factors and the glycemic control of patients with type 2 diabetes. METHODS: Participants were 304 patients with type 2 diabetes who were treated as outpatients at diabetes clinics. All participants were assessed for HbA1c and completed the following self-report psychological inventories: 1) Diabetes Treatment Satisfaction Questionnaire (DTSQ), 2) Problem Areas in Diabetes Survey (PAID), 3) Well-being Questionnaire 12 (W-BQ12), 4) Self-Esteem Scale (SES), 5) Social Support Scale, and 6) Self-Efficacy Scale. HbA1c was again measured one year later. The relationships between the psychosocial variables obtained by analysis of the psychological inventories and baseline or one-year follow-up HbA1c were determined. RESULTS: Baseline HbA1cwas significantly correlated with age, diet treatment regimen, number of microvascular complication of diabetes, and the total scores of DTSQ, W-BQ12, PAID, SES and the Self-Efficacy Scale. Hierarchical stepwise multiple regression revealed that significant predictors of baseline HbA1c were total DTSQ and PAID scores, along with age, diet treatment regimen, and number of microvascular complication of diabetes after adjustment for demographic, clinical and other psychosocial variables. Two hundred and ninety patients (95.4% of 304) were followed and assessed one year after baseline. Hierarchical stepwise multiple regression analysis showed the significant predictors of follow-up HbA1c to be total DTSQ and PAID scores, along with age and diet treatment regimen. However, the correlation between baseline and follow-up HbA1c was so high that the only other variable to retain significance was diet treatment regimen once baseline HbA1c was included in the regression of follow-up HbA1c. CONCLUSION: The DTSQ and the PAID predicted both current and future HbA1c to a similar and significant degree in patients with type 2 diabetes. PMID- 19298646 TI - Adenoviral vectors for highly selective gene expression in central serotonergic neurons reveal quantal characteristics of serotonin release in the rat brain. AB - BACKGROUND: 5-hydroxytryptamine (5 HT, serotonin) is one of the key neuromodulators in mammalian brain, but many fundamental properties of serotonergic neurones and 5 HT release remain unknown. The objective of this study was to generate an adenoviral vector system for selective targeting of serotonergic neurones and apply it to study quantal characteristics of 5 HT release in the rat brain. RESULTS: We have generated adenoviral vectors which incorporate a 3.6 kb fragment of the rat tryptophan hydroxylase-2 (TPH-2) gene which selectively (97% co-localisation with TPH-2) target raphe serotonergic neurones. In order to enhance the level of expression a two-step transcriptional amplification strategy was employed. This allowed direct visualization of serotonergic neurones by EGFP fluorescence. Using these vectors we have performed initial characterization of EGFP-expressing serotonergic neurones in rat organotypic brain slice cultures. Fluorescent serotonergic neurones were identified and studied using patch clamp and confocal Ca2+ imaging and had features consistent with those previously reported using post-hoc identification approaches. Fine processes of serotonergic neurones could also be visualized in un-fixed tissue and morphometric analysis suggested two putative types of axonal varicosities. We used micro-amperometry to analyse the quantal characteristics of 5 HT release and found that central 5 HT exocytosis occurs predominantly in quanta of approximately 28000 molecules from varicosities and approximately 34000 molecules from cell bodies. In addition, in somata, we observed a minority of large release events discharging on average approximately 800000 molecules. CONCLUSION: For the first time quantal release of 5 HT from somato-dendritic compartments and axonal varicosities in mammalian brain has been demonstrated directly and characterised. Release from somato-dendritic and axonal compartments might have different physiological functions. Novel vectors generated in this study open a host of new experimental opportunities and will greatly facilitate further studies of the central serotonergic system. PMID- 19298647 TI - A rapid and sensitive bioassay for the simultaneous measurement of multiple bone morphogenetic proteins. Identification and quantification of BMP4, BMP6 and BMP9 in bovine and human serum. AB - BACKGROUND: Bone morphogenetic proteins (BMPs) are pleiotropic members of the TGF beta superfamily which regulate many biological processes during development and adult tissue homeostasis and are implicated in the pathogenesis of a number of human diseases. Their involvement in both normal and aberrant physiology creates a need for rapid, sensitive and methodologically simple assays to evaluate their activity from a variety of biological samples. Previously alkaline phosphatase based assays, ELISA and luciferase based bioassays have been developed to evaluate either individual or total BMP activity. In this paper, we describe a highly sensitive, rapid and specific cell based assay for the simultaneous quantification of total and isoform specific BMP activity from biological samples. RESULTS: A C2C12 cell line stably transfected with a reporter plasmid consisting of the BMP response element (BRE) from the Id1 promoter fused to a luciferase reporter gene was generated. Exposure of this cell line to human recombinant BMP2, BMP4, BMP6, BMP7, BMP9 and BMP10 induced the expression of luciferase which was quantified using a luminometer. This assay was specific for BMP activity as the other TGF-beta superfamily members TGF-beta 1, Nodal and Mullerian Inhibiting Substance (MIS) did not induce the reporter. Pretreatment of samples with isoform specific BMP blocking antibodies coupled with isoform specific titration analysis allowed the simultaneous identification and quantification of BMP4, BMP6 and BMP9 in serum samples. CONCLUSION: The assay is rapid (<48 hours) and can be used to simultaneously measure isoform specific and total BMP activity in complex solutions. PMID- 19298648 TI - Spatiotemporal association of DNAJB13 with the annulus during mouse sperm flagellum development. AB - BACKGROUND: The sperm annulus is a septin-based fibrous ring structure connecting the midpiece and the principal piece of the mammalian sperm flagellum. Although ultrastructural abnormalities and functional importance of the annulus have been addressed in Sept4-null mutant mice and a subset of human patients with asthenospermia syndrome, little is known about how the structure is assembled and positioned to the midpiece-principal piece junction during mammalian sperm flagellum development. RESULTS: By performing immunofluorescence and biochemical approaches with antibodies against DNAJB13 and an annulus constituent SEPT4, we report here a spatiotemporal association of DNAJB13 with sperm annulus during mouse sperm flagellum development. DNAJB13 co-localized with SEPT4 to the annulus, and both were first able to be detected in step 9 spermatids. As spermiogenesis proceeded, the annular DNAJB13 immunosignal increased until the annulus reached the midpiece-principal piece junction, and then gradually disappeared from it in late spermiogenesis. In contrast, the SEPT4 immunosignal was relatively unaltered, and still present on annulus of mature spermatozoa. In Sept4-null mouse spermatids lacking the annulus structure, the annulus-like DNAJB13 immunosignal was still able to be detected, albeit weaker, at the neck region of the flagella. In vitro DNAJB13 was co-localized and interacted with SEPT4 directly. CONCLUSION: The direct interaction of DNAJB13 with SEPT4 in vitro and its spatiotemporal association with the annulus during sperm flagellum development, and even its annulus-like appearance in the annulus-deficient spermatids, suggest that DNAJB13 may be involved in assembling the annulus structure and positioning it towards the midpiece-principal piece junction. PMID- 19298649 TI - The epidemiology of gonorrhoea in Norway, 1993-2007: past victories, future challenges. AB - BACKGROUND: Gonorrhoea, a bacterial infection caused by Neisseria gonorrhoeae, has been increasing in several European countries, particularly among men who have sex with men (MSM) and teenagers. We describe the epidemiology of gonorrhoea in Norway in the recent 15 years in order to guide recommendations on the diagnosis, treatment and prevention of gonorrhoea. An evaluation of the Norwegian Surveillance System for Communicable Diseases (MSIS) in 1994, involving GPs and microbiological laboratories, suggested that the system has a high coverage, capturing over 90% of patients diagnosed with gonorrhoea. METHODS: Using MSIS data on gonorrhoea cases we analysed specific trends by route of transmission, age, gender, anatomical sampling site, antimicrobial resistance and travel history from 1993-2007 and, to focus on more recent trends, from 2003-2007. MSM and heterosexual cases were defined by route of transmission. RESULTS: From 1993 to 2007, 3601 gonorrhoea cases were reported. MSM cases increased from 10 in 1994 to 109 cases in 2004. From 2003-2007, the incidence of gonorrhoea was 5.4/100,000 person-years (95%CI: 4.9-6.0). Over these five years, MSM accounted for an average of 80 cases per year, of which 69% were infected by casual partners. In the same period, 98% of heterosexually infected had a positive swab from urethra only and only two (0.3%) from the pharynx. Only one woman (0.5%) was positive from the rectum. From 1993 - 2007, antimicrobial resistance results were reported for 3325 N. gonorrhoeae isolates (98% of cultured samples). The proportion resistant to quinolone has risen from 3% in 1995 to 47% in 2007, with 81% of the latter isolated from patients infected in Asia. CONCLUSION: The overall incidence of gonorrhoea in Norway remains low, but the increasing number of MSM cases calls for new, more effective approaches to prevention. Infections originating from abroad represent a constant risk of importing antimicrobial resistant N. gonorrhoeae. Due to the prevalence of quinolone resistant N. gonorrhoeae in Norway, third-generation cephalosporins should replace quinolones as the first choice in treatment guidelines. We advocate antimicrobial susceptibility testing for all cases and recommend taking samples for culture from all exposed anatomical sites. PMID- 19298650 TI - Measuring cough severity: Perspectives from the literature and from patients with chronic cough. AB - BACKGROUND: In order to assess severity of cough from patients' perspectives and capture the effects of treatment in clinical trials, a measurement tool must show evidence of validity and reliability. The purpose of this study was to characterize cough severity from patients' perspectives as the initial step in the development of a new patient-reported outcome (PRO) measure for use in clinical trials. METHODS: This focus groups study included patients with clinician confirmed chronic cough recruited from a large internal medicine clinic in the US. A semi-structured focus group guide was designed to elicit information about patients' experiences with cough severity and their characterization of symptoms. The focus group data were coded to identify concepts and terminology of cough severity. RESULTS: Three focus groups were conducted [n = 22; 6 male; mean age 66.1 (+/- 12.9)]. Etiology included GERD, asthma, bronchitis, post-nasal drip, and other. Three domains of cough severity were identified: frequency, intensity, and disruption. In addition to a single cough, participants in all focus groups described coughing in uncontrollable paroxysms they called "fits," "bouts," "spells," or "episodes." The urge to cough, described as an important sign of impending cough, was considered a component of cough frequency. Participants also described daytime activity and nighttime sleep disruption as an indication of cough severity. Finally, participants described variability in cough severity. CONCLUSION: Results suggest that patients describe cough severity in terms of frequency, intensity, and disruptiveness, indicating these 3 domains should be addressed when evaluating cough severity and outcomes of treatment. PMID- 19298651 TI - High Erk-1 activation and Gadd45a expression as prognostic markers in high risk pediatric haemolymphoproliferative diseases. AB - Studies on activated cell-signaling pathways responsible for neoplastic transformation are numerous in solid tumors and in adult leukemias. Despite of positive results in the evolution of pediatric hematopoietic neoplasias, there are some high-risk subtypes at worse prognosis. The aim of this study was to asses the expression and activation status of crucial proteins involved in cell signaling pathways in order to identify molecular alterations responsible for the proliferation and/or escape from apoptosis of leukemic blasts. The quantitative and qualitative expression and activation of Erk-1, c-Jun, Caspase8, and Gadd45a was analyzed, by immunocytochemical (ICC) and western blotting methods, in bone marrow blasts of 72 patients affected by acute myeloid leukemia (AML), T-cell acute lymphoblastic leukemia (ALL) and stage IV non-Hodgkin Lymphoma (NHL). We found an upregulation of Erk-1, Caspase8, c-Jun, and Gadd45a proteins with a constitutive activation in 95.8%, 91.7%, 86.2%, 83.4% of analyzed specimens, respectively. It is worth noting that all AML patients showed an upregulation of all proteins studied and the high expression of GADD45a was associated to the lowest DFS median (p = 0.04). On univariate analysis, only Erk-1 phosphorylation status was found to be correlated with a significantly shorter 5-years DFS in all disease subgroups (p = 0.033) and the lowest DFS median in ALL/NHL subgroup (p = 0.04). Moreover, the simultaneous activation of multiple kinases, as we found for c-Jun and Erk-1 (r = 0.26; p = 0.025), might synergistically enhance survival and proliferation potential of leukemic cells. These results demonstrate an involvement of these proteins in survival of blast cells and, consequently, on relapse percentages of the different subgroups of patients. PMID- 19298652 TI - Interferon-gamma coordinates CCL3-mediated neutrophil recruitment in vivo. AB - BACKGROUND: We have shown previously that acute infection with the respiratory pathogen, pneumonia virus of mice (PVM), results in local production of the proinflammatory chemokine, CCL3, and that neutrophil recruitment in response to PVM infection is reduced dramatically in CCL3 -/- mice. RESULTS: In this work, we demonstrate that CCL3-mediated neutrophil recruitment is coordinated by interferon-gamma (IFNgamma). Neutrophil recruitment in response to PVM infection was diminished five-fold in IFNgamma receptor gene-deleted mice, although neutrophils from IFNgammaR -/- mice expressed transcripts for the CCL3 receptor, CCR1 and responded functionally to CCL3 ex vivo. Similarly, in the absence of PVM infection, CCL3 overexpression alone could not elicit neutrophil recruitment in the absence of IFNgamma. Interestingly, although supplemental IFNgamma restored neutrophil recruitment and resulted in a sustained weight loss among CCL3 overexpressing IFNgamma -/- mice, CCL3-mediated neutrophil recruitment alone did not result in the pulmonary edema or respiratory failure characteristic of severe viral infection, suggesting that CCL3 and IFN-gamma together are sufficient to promote neutrophil recruitment but not pathologic activation. CONCLUSION: Our findings reveal a heretofore unrecognized hierarchical interaction between the IFNgamma and CCL3, which demonstrate that IFNgamma is crucial for CCL3-mediated neutrophil recruitment in vivo. PMID- 19298653 TI - Factors associated with self-reported first sexual intercourse in Scottish adolescents. AB - BACKGROUND: There is continuing concern about high pregnancy rates and increasing numbers of sexually transmitted infections being detected in Scottish adolescents. Consistent evidence about factors associated with risky sexual behaviours, including early first sexual intercourse, may help to identify adolescents at risk and help improve interventions. This study aimed to provide detailed analysis of the evidence of the associations between individual factors and early sexual intercourse using cross-sectional questionnaire data from 4,379 Scottish adolescents who participated in a sexual health intervention evaluation. FINDINGS: Multivariate secondary analysis showed that aspects of family and school life such as decreasing parental monitoring (OR 1.45, 95% CI 1.24-1.70) and decreasing enjoyment of school (OR 2.55, 95% CI 2.15-3.03) were associated with reporting previous sexual intercourse. Furthermore, females were more likely to report previous sexual intercourse than males (OR 1.48, 95% CI 1.14-1.91). Several factors commonly used to inform sexual health intervention design, such as socioeconomic status, self-esteem and religion, were not independently associated. CONCLUSION: These results contribute to the evidence base for the association of several factors with early initiation of sexual activity. The findings suggest that interventions aiming to delay first intercourse may need to consider targeting aspects of individuals' connection to their school and family. Furthermore, the results do not support the need to consider socio-economic background, religion or self-esteem of the individuals in intervention design. PMID- 19298655 TI - Peroxisome proliferator-activated receptor-gamma protects ERBB2-positive breast cancer cells from palmitate toxicity. AB - INTRODUCTION: Accumulation of fatty acids and neutral lipids in nonadipose tissues is cytotoxic. We recently showed that ERBB2-positive breast cancer cells produce significantly high amounts of fats, because of overexpression of the peroxisome proliferator-activated receptor (PPAR)gamma-binding protein and the nuclear receptor NR1D1 (nuclear receptor subfamily 1, group D, member 1; Rev erbalpha). These genes upregulate de novo fatty acid synthesis, which is a critical pathway for the energy production and survival of these cells. NR1D1 and PPARgamma-binding protein are functionally related to PPARgamma, a well established positive regulator of adipogenesis and lipid storage. METHODS: The effects of GW9662 and exogenously added palmitate on breast cells (BT474, MDA-MB 361, MCF-7, and human mammary epithelial cells) in monolayer culture were assessed. Mass spectrometric quantitation of fatty acids and fluorescence-based high content microscopy assays of cell growth, apoptosis, triglyceride storage and reactive oxygen species production were used. RESULTS: ERBB2-positive breast cancer cells are more sensitive to inhibition of PPARgamma activity by the antagonist GW9662. PPARgamma inhibition results in increased levels of total fats in the cells, mostly because of increased amounts of palmitic and stearic unsaturated acids. Administration of exogenous palmitate is lethal to ERBB2 positive but not to ERBB2-negative cells. GW9662 exacerbates the effects of palmitate addition on BT474 and MDA-MB-361 cells, but it has no significant effect on MCF-7 and human mammary epithelial cells. Palmitate administration results in a fivefold to tenfold greater increase in fat stores in ERBB2-negative cells compared with ERBB2-positive cells, which suggests that the ERBB2-positive cells have maximized their ability to store fats and that additional palmitate is toxic to these cells. Both PPARgamma inhibition and palmitate administration result in increased reactive oxygen species production in BT474 cells. The cell death that results from this treatment can be counteracted by the antioxidant N acetyl cysteine. CONCLUSIONS: Our findings indicate that PPARgamma activity enables ERBB2-positive breast cancer cells, which produce high levels of fat, to convert fatty acids to triglycerides, allowing these cells to avert the cell death that results from lipotoxicity. Endogenous palmitate toxicity represents a genetically based property of ERBB2-positive breast cancer that can be exploited for therapeutic intervention. PMID- 19298656 TI - A demographic and epidemiological study of a Mexican chiropractic college public clinic. AB - BACKGROUND: Descriptive studies of chiropractic patients are not new, several have been performed in the U.S., Australia, Canada, and Europe. None have been performed in a Latin American country. The purpose of this study is to describe the patients who visited a Mexican chiropractic college public clinic with respect to demographics and clinical characteristics. METHODS: This study was reviewed and approved by the IRB of Parker College of Chiropractic and the Universidad Estatal del Valle de Ecatepec (UNEVE). Five hundred patient files from the UNEVE public clinic from May 2005 to May 2007 were selected from an approximate total number of 3,700. Information was collected for demographics, chief complaints, associated complaints, and previous care sought. RESULTS: The sample comprised 306 (61.2%) female. Most files (44.2%) were in the age range of 40-59 years (mean of 43.4 years). The most frequent complaints were lumbar pain (29.2%) and extremity pain (28.0%), most commonly the knee. Most (62.0%) described their complaints as greater than one year. Trauma (46.6%) was indicated as the initial cause. Mean VAS score was 6.26/10 with 20% rated at 8/10. CONCLUSION: Demographic results compared closer to studies conducted with private clinicians (females within the ages of 40-59). The primary complaint and duration was similar to previous studies (low back pain and chronic), except in this population the cause was usually initiated by trauma. The most striking features were the higher number of extremity complaints and the marked increased level of VAS score (20% rated as 8/10). PMID- 19298657 TI - Relating perturbation magnitude to temporal gene expression in biological systems. AB - BACKGROUND: Most transcriptional activity is a result of environmental variability. This cause (environment) and effect (gene expression) relationship is essential to survival in any changing environment. The specific relationship between environmental perturbation and gene expression - and stability of the response - has yet to be measured in detail. We describe a method to quantitatively relate perturbation magnitude to response at the level of gene expression. We test our method using Saccharomyces cerevisiae as a model organism and osmotic stress as an environmental stress. RESULTS: Patterns of gene expression were measured in response to increasing sodium chloride concentrations (0, 0.5, 0.7, 1.0, and 1.2 M) for sixty genes impacted by osmotic shock. Expression of these genes was quantified over five time points using reverse transcriptase real-time polymerase chain reaction. Magnitudes of cumulative response for specific pathways, and the set of all genes, were obtained by combining the temporal response envelopes for genes exhibiting significant changes in expression with time. A linear relationship between perturbation magnitude and response was observed for the range of concentrations studied. CONCLUSION: This study develops a quantitative approach to describe the stability of gene response and pathways to environmental perturbation and illustrates the utility of this approach. The approach should be applicable to quantitatively evaluate the response of organisms via the magnitude of response and stability of the transcriptome to environmental change. PMID- 19298658 TI - Qualitative assessment of innovations in healthcare provision. AB - BACKGROUND: The triad of quality, innovation and economic restraint is as important in health care as it is in the business world. There are many proposals for the assessment of quality and of economic restraints in health care but only a few address assessment of innovations. We propose a strategy and new structures to standardize the description of health care innovations and to quantify them. DISCUSSION: Strategy and structure are based on the assumption that in the early phase of an innovation only data on the feasibility and possibly on the efficacy or effectiveness of an innovation can be expected. From the patient's perspective, benefit resulting from an innovation can be confirmed only in a later phase of development. Early indicators of patient's benefit will be surrogate parameters which correlate only weakly with the desired endpoints. After the innovation has been in use, there will be more evidence on correlations between surrogate parameters and the desired endpoints to provide evidence of the patient benefit. From an administrative perspective, this evidence can be considered in decisions about public financing. Different criteria are proposed for the assessment of innovations in prevention, diagnosis and therapy. For decisions on public financing a public fund for innovations may be helpful. Depending on the phase of innovation risk sharing models are proposed between manufacturers, private insurers and public funding. SUMMARY: Potential for patient benefit is always uncertain during early stages of innovations. This uncertainty decreases with increasing information on the effects of the innovation. Information about an innovation can be quantified, categorized and integrated into rational economic decisions. PMID- 19298659 TI - The short-term safety and efficacy of fluoxetine in depressed adolescents with alcohol and cannabis use disorders: a pilot randomized placebo-controlled trial. AB - BACKGROUND: The objective of this study was to examine whether fluoxetine was superior to placebo in the acute amelioration of depressive symptomatology in adolescents with depressive illness and a comorbid substance use disorder. METHODS: Eligible subjects ages 12-17 years with either a current major depressive disorder (MDD) or a depressive disorder that were also suffering from a comorbid substance-related disorder were randomized to receive either fluoxetine or placebo in this single site, 8-week double-blind, placebo controlled study. The primary outcome analysis was a random effects mixed model for repeated measurements of Children's Depression Rating Scale-Revised (CDRS-R) scores compared between treatment groups across time. RESULTS: An interim analysis was performed after 34 patients were randomized. Based on the results of a futility analysis, study enrollment was halted. Twenty-nine males and 5 females were randomized to receive fluoxetine (n = 18) or placebo (n = 16). Their mean age was 16.5 (1.1) years. Overall, patients who received fluoxetine and placebo had a reduction in CDRS-R scores. However, there was no significant difference in mean change in CDRS-R total score in those subjects treated with fluoxetine and those who received placebo (treatment difference = 0.19, S.E. = 0.58, F = 0.14, p = .74). Furthermore, there was not a significant difference in rates of positive urine drug toxicology results between treatment groups at any post-randomization visit (F = 0.22, df = 1, p = 0.65). The main limitation of this study is its modest sample size and resulting low statistical power. Other significant limitations to this study include, but are not limited to, the brevity of the trial, high placebo response rate, limited dose range of fluoxetine, and the inclusion of youth who met criteria for depressive disorders other than MDD. CONCLUSION: Fluoxetine was not superior to placebo in alleviating depressive symptoms or in decreasing rates of positive drug screens in the acute treatment of adolescents with depression and a concomitant substance use disorder. PMID- 19298660 TI - Tumor necrosis factor-alpha induced expression of matrix metalloproteinase-9 through p21-activated kinase-1. AB - BACKGROUND: Expressed in embryonic development, matrix metalloprotein-9 (MMP-9) is absent in most of developed adult tissues, but recurs in inflammation during tissue injury, wound healing, tumor formation and metastasis. Expression of MMP-9 is tightly controlled by extracellular cues including pro-inflammatory cytokines and extracellular matrix (ECM). While the pathologic functions of MMP-9 are evident, the intracellular signaling pathways to control its expression are not fully understood. In this study we investigated mechanism of cytokine induced MMP 9 with particular emphasis on the role of p21-activated-kinase-1 (PAK1) and the down stream signaling. RESULTS: In response to TNF-alpha or IL-1alpha, PAK1 was promptly activated, as characterized by a sequential phosphorylation, initiated at threonine-212 followed by at threonine-423 in the activation loop of the kinase, in human skin keratinocytes, dermal fibroblasts, and rat hepatic stellate cells. Ectopic expression of PAK1 variants, but not p38 MAP kinase, impaired the TNF-alpha-induced MMP-9 expression, while other MMPs such as MMP-2, -3 and -14 were not affected. Activation of Jun N-terminal kinase (JNK) and NF-kappaB has been demonstrated to be essential for MMP-9 expression. Expression of inactive PAK1 variants impaired JNK but not NF-kappaB activation, which consequently suppressed the 5'-promoter activities of the MMP-9 gene. After the cytokine induced phosphorylation, both ectopically expressed and endogenous PAK1 proteins were promptly accumulated even in the condition of suppressing protein synthesis, suggesting the PAK1 protein is stabilized upon TNF-alpha stimulation. Stabilization of PAK1 protein by TNF-alpha treatment is independent of the kinase catalytic activity and p21 GTPase binding capacities. In contrast to epithelial cells, mesenchymal cells require 3-dimensional type-I collagen in response to TNF alpha to massively express MMP-9. The collagen effect is mediated, in part, by boost JNK activation in a way to cooperate the cytokine signaling. CONCLUSION: We identified a novel mechanism for MMP-9 expression in response to injury signals, which is mediated by PAK1 activation and stabilization leading JNK activation. PMID- 19298661 TI - Redundancy of myostatin and growth/differentiation factor 11 function. AB - BACKGROUND: Myostatin (Mstn) and growth/differentiation factor 11 (Gdf11) are highly related transforming growth factor beta (TGFbeta) family members that play important roles in regulating embryonic development and adult tissue homeostasis. Despite their high degree of sequence identity, targeted mutations in these genes result in non-overlapping phenotypes affecting distinct biological processes. Loss of Mstn in mice causes a doubling of skeletal muscle mass while loss of Gdf11 in mice causes dramatic anterior homeotic transformations of the axial skeleton, kidney agenesis, and an increase in progenitor cell number in several tissues. In order to investigate the possible functional redundancy of myostatin and Gdf11, we analyzed the effect of eliminating the functions of both of these signaling molecules. RESULTS: We show that Mstn-/- Gdf11-/- mice have more extensive homeotic transformations of the axial skeleton than Gdf11-/- mice in addition to skeletal defects not seen in single mutants such as extra forelimbs. We also show that deletion of Gdf11 specifically in skeletal muscle in either Mstn+/+ or Mstn-/- mice does not affect muscle size, fiber number, or fiber type. CONCLUSION: These results provide evidence that myostatin and Gdf11 have redundant functions in regulating skeletal patterning in mice but most likely not in regulating muscle size. PMID- 19298662 TI - The prevalence of BRCA1 mutations among young women with triple-negative breast cancer. AB - BACKGROUND: Molecular screening for BRCA1 and BRCA2 mutations is now an established component of risk evaluation and management of familial breast cancer. Features of hereditary breast cancer include an early age-of-onset and over-representation of the 'triple-negative' phenotype (negative for estrogen receptor, progesterone-receptor and HER2). The decision to offer genetic testing to a breast cancer patient is usually based on her family history, but in the absence of a family history of cancer, some women may qualify for testing based on the age-of-onset and/or the pathologic features of the breast cancer. METHODS: We studied 54 women who were diagnosed with high-grade, triple-negative invasive breast cancer at or before age 40. These women were selected for study because they had little or no family history of breast or ovarian cancer and they did not qualify for genetic testing using conventional family history criteria. BRCA1 screening was performed using a combination of fluorescent multiplexed-PCR analysis, BRCA1 exon-13 6 kb duplication screening, the protein truncation test (PTT) and fluorescent multiplexed denaturing gradient gel electrophoresis (DGGE). All coding exons of BRCA1 were screened. The two large exons of BRCA2 were also screened using PTT. All mutations were confirmed with direct sequencing. RESULTS: Five deleterious BRCA1 mutations and one deleterious BRCA2 mutation were identified in the 54 patients with early-onset, triple-negative breast cancer (11%). CONCLUSION: Women with early-onset triple-negative breast cancer are candidates for genetic testing for BRCA1, even in the absence of a family history of breast or ovarian cancer. PMID- 19298663 TI - Two-dimensional gel proteome reference map of human small intestine. AB - BACKGROUND: The small intestine is an important human organ that plays a central role in many physiological functions including digestion, absorption, secretion and defense. Duodenal pathologies include, for instance, the ulcer associated to Helicobacter Pylori infection, adenoma and, in genetically predisposed individuals, celiac disease. Alterations in the bowel reduce its capability to absorb nutrients, minerals and fat-soluble vitamins. Anemia and osteopenia or osteoporosis may develop as a consequence of vitamins malabsorption. Adenoma is a benign tumor that has the potential to become cancerous. Adult celiac disease patients present an overall risk of cancer that is almost twice than that found in the general population. These disease processes are not completely known.To date, a two dimensional (2D) reference map of proteins expressed in human duodenal tissue is not yet available: the aim of our study was to characterize the 2D protein map, and to identify proteins of duodenal mucosa of adult individuals without duodenal illness, to create a protein database. This approach, may be useful for comparing similar protein samples in different laboratories and for the molecular characterization of intestinal pathologies without recurring to the use of surgical material. RESULTS: The enrolled population comprised five selected samples (3 males and 2 females, aged 19 to 42), taken from 20 adult subjects, on their first visit at the gastroenterology unit for a suspected celiac disease, who did not turn to be affected by any duodenal pathology after gastrointestinal and histological evaluations. Proteins extracted from the five duodenal mucosal specimens were singly separated by 2D gel electrophoresis. After image analysis of each 2D gel, 179 protein spots, representing 145 unique proteins, from 218 spots tested, were successfully identified by MALDI-TOF ms analysis. Normalized volumes, for each protein, have been reported for every gel. Proteins have been grouped according to their biological/metabolic functions. CONCLUSION: This study represents to date the first detailed and reproducible 2D protein map of human duodenum. Spots identifications, reported in a database, will be helpful to identify the variability in protein expression levels, in isoforms expression, or in post translational modifications associated to pathology or to a therapy. PMID- 19298665 TI - Ascorbic acid pre-treated quartz stimulates TNF-alpha release in RAW 264.7 murine macrophages through ROS production and membrane lipid peroxidation. AB - BACKGROUND: Inhalation of crystalline silica induces a pulmonary fibrotic degeneration called silicosis caused by the inability of alveolar macrophages to dissolve the crystalline structure of phagocytosed quartz particles. Ascorbic acid is capable of partially dissolving quartz crystals, leading to an increase of soluble silica concentration and to the generation of new radical sites on the quartz surface. The reaction is specific for the crystalline forms of silica. It has been already demonstrated an increased cytotoxicity and stronger induction of pro-inflammatory cyclooxygenase-2 (COX-2) by ascorbic acid pre-treated quartz (QA) compared to untreated quartz (Q) in the murine macrophage cell line RAW 264.7. METHODS: Taking advantage of the enhanced macrophage response to QA as compared to Q particles, we investigated the first steps of cell activation and the contribution of early signals generated directly from the plasma membrane to the production of TNF-alpha, a cytokine that activates both inflammatory and fibrogenic pathways. RESULTS: Here we demonstrate that TNF-alpha mRNA synthesis and protein secretion are significantly increased in RAW 264.7 macrophages challenged with QA as compared to Q particles, and that the enhanced response is due to an increase of intracellular ROS. Plasma membrane-particle contact, in the absence of phagocytosis, is sufficient to trigger TNF-alpha production through a mechanism involving membrane lipid peroxidation and this appears to be even more detrimental to macrophage survival than particle phagocytosis itself. CONCLUSION: Taken together these data suggest that an impairment of pulmonary macrophage phagocytosis, i.e. in the case of alcoholic subjects, could potentiate lung disease in silica-exposed individuals. PMID- 19298666 TI - Calculation of health expectancies with administrative data for North Rhine Westphalia, a Federal State of Germany, 1999-2005. AB - OBJECTIVES: The main objectives of this study were to prove the feasibility of health expectancy analyses with regional administrative health statistics and to explore the utility of the calculated health expectancies in describing the health state of the population living in North Rhine-Westphalia, a Federal State of Germany. MATERIALS AND METHODS: Administrative population and mortality data as well as health data on disability and long-term care provided by public services were used to calculate: a) the life expectancy and b) the health expectancies Severe-Disability-Free Life Expectancy (SDFLE) and Long-Term-Care Free Life Expectancy (LTCFLE) from 1999 to 2005. Calculations were done using the Sullivan method. RESULTS: SDFLE at birth was 69.9 years (males 66.2 and females 73.2 years) in 1999 and it increased to 71.7 years (males 68.6 and females 74.7 years) in 2005. The proportion of the SDFLE on the total life expectancy at birth was 89.8% (males 88.6 and females 90.8%) in 1999 and 90.7% (males 89.8 and females 91.4%) in 2005.LTCFLE at birth was 75.3 years (males 73.1 and females 77.5 years) in 1999 and it increased to 76.6 years (males 74.7 and females 78.6 years) in 2005. The proportion of the LTCFLE on the total life expectancy at birth was 96.8% (males 97.8 and females 96.1%) in 1999 and 96.8% (males 97.8 and females 96.2%) in 2005. DISCUSSION AND CONCLUSION: Both health expectancies indicate an improvement in the quantity as well as in the quality of healthy life for the population living in North Rhine Westphalia and therefore suggest a compression of morbidity from 1999 to 2005. The findings however have several limitations in their sensitivity, since we applied dichotomous valuations to the health states. In addition, the results are restricted to comparisons over time because the morbidity concepts do not allow for comparisons with populations other than the German one. Refined calculations with other summary measures of population health and with health data on other morbidity concepts are therefore reasonable. PMID- 19298667 TI - Digital PCR provides sensitive and absolute calibration for high throughput sequencing. AB - BACKGROUND: Next-generation DNA sequencing on the 454, Solexa, and SOLiD platforms requires absolute calibration of the number of molecules to be sequenced. This requirement has two unfavorable consequences. First, large amounts of sample-typically micrograms-are needed for library preparation, thereby limiting the scope of samples which can be sequenced. For many applications, including metagenomics and the sequencing of ancient, forensic, and clinical samples, the quantity of input DNA can be critically limiting. Second, each library requires a titration sequencing run, thereby increasing the cost and lowering the throughput of sequencing. RESULTS: We demonstrate the use of digital PCR to accurately quantify 454 and Solexa sequencing libraries, enabling the preparation of sequencing libraries from nanogram quantities of input material while eliminating costly and time-consuming titration runs of the sequencer. We successfully sequenced low-nanogram scale bacterial and mammalian DNA samples on the 454 FLX and Solexa DNA sequencing platforms. This study is the first to definitively demonstrate the successful sequencing of picogram quantities of input DNA on the 454 platform, reducing the sample requirement more than 1000 fold without pre-amplification and the associated bias and reduction in library depth. CONCLUSION: The digital PCR assay allows absolute quantification of sequencing libraries, eliminates uncertainties associated with the construction and application of standard curves to PCR-based quantification, and with a coefficient of variation close to 10%, is sufficiently precise to enable direct sequencing without titration runs. PMID- 19298668 TI - Improved analysis of bacterial CGH data beyond the log-ratio paradigm. AB - BACKGROUND: Existing methods for analyzing bacterial CGH data from two-color arrays are based on log-ratios only, a paradigm inherited from expression studies. We propose an alternative approach, where microarray signals are used in a different way and sequence identity is predicted using a supervised learning approach. RESULTS: A data set containing 32 hybridizations of sequenced versus sequenced genomes have been used to test and compare methods. A ROC-analysis has been performed to illustrate the ability to rank probes with respect to Present/Absent calls. Classification into Present and Absent is compared with that of a gaussian mixture model. CONCLUSION: The results indicate our proposed method is an improvement of existing methods with respect to ranking and classification of probes, especially for multi-genome arrays. PMID- 19298669 TI - Analysis of tarantula skeletal muscle protein sequences and identification of transcriptional isoforms. AB - BACKGROUND: Tarantula has been used as a model system for studying skeletal muscle structure and function, yet data on the genes expressed in tarantula muscle are lacking. RESULTS: We constructed a cDNA library from Aphonopelma sp. (Tarantula) skeletal muscle and got 2507 high-quality 5'ESTs (expressed sequence tags) from randomly picked clones. EST analysis showed 305 unigenes, among which 81 had more than 2 ESTs. Twenty abundant unigenes had matches to skeletal muscle related genes including actin, myosin, tropomyosin, troponin-I, T and C, paramyosin, muscle LIM protein, muscle protein 20, a-actinin and tandem Ig/Fn motifs (found in giant sarcomere-related proteins). Matches to myosin light chain kinase and calponin were also identified. These results support the existence of both actin-linked and myosin-linked regulation in tarantula skeletal muscle. We have predicted full-length as well as partial cDNA sequences both experimentally and computationally for myosin heavy and light chains, actin, tropomyosin, and troponin-I, T and C, and have deduced the putative peptides. A preliminary analysis of the structural and functional properties was also carried out. Sequence similarities suggested multiple isoforms of most myofibrillar proteins, supporting the generality of multiple isoforms known from previous muscle sequence studies. This may be related to a mix of muscle fiber types. CONCLUSION: The present study serves as a basis for defining the transcriptome of tarantula skeletal muscle, for future in vitro expression of tarantula proteins, and for interpreting structural and functional observations in this model species. PMID- 19298670 TI - Dietary supplementation of mannan-oligosaccharide enhances neonatal immune responses in chickens during natural exposure to Eimeria spp. AB - BACKGROUND: Control and eradication of intestinal infections caused by protozoa are important biomedical challenges worldwide. Prophylactic control of coccidiosis has been achieved with the use of anticoccidial drugs; however, the increase in anticoccidial resistance has raised concerns about the need for new alternatives for the control of coccidial infections. In fact, new strategies are needed to induce potent protective immune responses in neonatal individuals. METHODS: The effects of a dietary supplementation of mannan-oligosaccharide (yeast cell wall; YCW) on the local, humoral and cell-mediated immune responses, and intestinal replication of coccidia were evaluated in a neonatal animal model during natural exposure to Eimeria spp. A total of 840 one-day-old chicks were distributed among four dietary regimens: A) Control diet (no YCW) plus anticoccidial vaccine); B) Control diet plus coccidiostat; C) YCW diet plus anticoccidial vaccination; and D) YCW diet plus coccidiostat. Weight gain, feed consumption and immunological parameters were examined within the first seven weeks of life. RESULTS: Dietary supplementation of 0.05% of YCW increased local mucosal IgA secretions, humoral and cell-mediated immune responses, and reduced parasite excretion in feces. CONCLUSION: Dietary supplementation of yeast cell wall in neonatal animals can enhance the immune response against coccidial infections. The present study reveals the potential of YCW as adjuvant for modulating mucosal immune responses. PMID- 19298671 TI - Information processing in the transcriptional regulatory network of yeast: functional robustness. AB - BACKGROUND: Gene networks are considered to represent various aspects of molecular biological systems meaningfully because they naturally provide a systems perspective of molecular interactions. In this respect, the functional understanding of the transcriptional regulatory network is considered as key to elucidate the functional organization of an organism. RESULTS: In this paper we study the functional robustness of the transcriptional regulatory network of S. cerevisiae. We model the information processing in the network as a first order Markov chain and study the influence of single gene perturbations on the global, asymptotic communication among genes. Modification in the communication is measured by an information theoretic measure allowing to predict genes that are 'fragile' with respect to single gene knockouts. Our results demonstrate that the predicted set of fragile genes contains a statistically significant enrichment of so called essential genes that are experimentally found to be necessary to ensure vital yeast. Further, a structural analysis of the transcriptional regulatory network reveals that there are significant differences between fragile genes, hub genes and genes with a high betweenness centrality value. CONCLUSION: Our study does not only demonstrate that a combination of graph theoretical, information theoretical and statistical methods leads to meaningful biological results but also that such methods allow to study information processing in gene networks instead of just their structural properties. PMID- 19298672 TI - Generation in vivo of peptide-specific cytotoxic T cells and presence of regulatory T cells during vaccination with hTERT (class I and II) peptide-pulsed DCs. AB - BACKGROUND: Optimal techniques for DC generation for immunotherapy in cancer are yet to be established. Study aims were to evaluate: (i) DC activation/maturation milieu (TNF-alpha +/- IFN-alpha) and its effects on CD8+ hTERT-specific T cell responses to class I epitopes (p540 or p865), (ii) CD8+ hTERT-specific T cell responses elicited by vaccination with class I alone or both class I and II epitope (p766 and p672)-pulsed DCs, prepared without IFN-alpha, (iii) association between circulating T regulatory cells (Tregs) and clinical responses. METHODS: Autologous DCs were generated from 10 patients (HLA-0201) with advanced cancer by culturing CD14+ blood monocytes in the presence of GM-CSF and IL-4 supplemented with TNF-alpha [DCT] or TNF-alpha and IFN-alpha [DCTI]. The capacity of the DCs to induce functional CD8+ T cell responses to hTERT HLA-0201 restricted nonapeptides was assessed by MHC tetramer binding and peptide-specific cytotoxicity. Each DC preparation (DCT or DCTI) was pulsed with only one type of hTERT peptide (p540 or p865) and both preparations were injected into separate lymph node draining regions every 2-3 weeks. This vaccination design enabled comparison of efficacy between DCT and DCTI in generating hTERT peptide specific CD8+ T cells and comparison of class I hTERT peptide (p540 or p865)-loaded DCT with or without class II cognate help (p766 and p672) in 6 patients. T regulatory cells were evaluated in 8 patients. RESULTS: (i) DCTIs and DCTs, pulsed with hTERT peptides, were comparable (p = 0.45, t-test) in inducing peptide-specific CD8+ T cell responses. (ii) Class II cognate help, significantly enhanced (p < 0.05, t-test) peptide-specific CD8+T cell responses, compared with class I pulsed DCs alone. (iii) Clinical responders had significantly lower (p < 0.05, Mann Whitney U test) T regs, compared with non-responders. 4/16 patients experienced partial but transient clinical responses during vaccination. Vaccination was well tolerated with minimal toxicity. CONCLUSION: Addition of IFN-alpha to ex vivo monocyte-derived DCs, did not significantly enhance peptide-specific T cell responses in vivo, compared with TNF-alpha alone. Class II cognate help significantly augments peptide-specific T cell responses. Clinically favourable responses were seen in patients with low levels of circulating T regs. PMID- 19298673 TI - Assessment of anti-cyclic citrullinated peptide in psoriatic arthritis. AB - BACKGROUND: Anti-cyclic citrullinated peptides (anti-CCP) are highly specific diagnostic and prognostic markers for rheumatoid arthritis (RA). They have been also found in psoriatic arthritis (PsA), with controversies as regards clinical and radiological associations. The current study assessed anti-CCP in PsA and determined its clinical and radiological associations. METHODS: Four groups contributed to this study. 40 PsA, 40 psoriasis without arthritis, 40 RA and 40 healthy controls. They were tested for anti-CCP. Clinical and radiological data were collected and statistically compared between anti-CCP-positive and -negative PsA patients. RESULTS: Seven PsA (17.5%) and 34 RA (85%) were seropositive for anti-CCP. Patients of other groups were anti-CCP-negative. Regarding anti-CCP concentrations, highly significant difference existed between different groups and between anti-CCP-positive and -negative PsA. Significantly higher numbers of involved, swollen and tender joints, deformities and functional impairment of peripheral joints and radiological changes were found in anti-CCP-positive PsA. CONCLUSION: Anti-CCP may be found in PsA and are associated with higher number of involved, swollen and tender joints, with deformities and functional impairment of peripheral joints and with erosive arthritis. PMID- 19298675 TI - Evolving DNA motifs to predict GeneChip probe performance. AB - BACKGROUND: Affymetrix High Density Oligonuclotide Arrays (HDONA) simultaneously measure expression of thousands of genes using millions of probes. We use correlations between measurements for the same gene across 6685 human tissue samples from NCBI's GEO database to indicated the quality of individual HG-U133A probes. Low correlation indicates a poor probe. RESULTS: Regular expressions can be automatically created from a Backus-Naur form (BNF) context-free grammar using strongly typed genetic programming. CONCLUSION: The automatically produced motif is better at predicting poor DNA sequences than an existing human generated RE, suggesting runs of Cytosine and Guanine and mixtures should all be avoided. PMID- 19298674 TI - Anti-idiotypic antibodies: a new approach in prion research. AB - BACKGROUND: In certain cases, anti-idiotypic antibodies that recognize an antigen combining site of an antibody can mimic the structure and/or function of certain nominal antigens. This feature makes them particularly useful if conventional experimental approaches fail to fulfil expectations, especially when the molecule of interest is infectious, toxic or difficult to isolate and purify. We suggest the application of an anti-idiotype concept to the field of prion biology, with the aim of evoking a humoral immune response against the pathological isoform of the prion protein (PrPSc). Different ways to induce anti-idiotypic responses were studied in mice and chickens using various forms of V5B2, a PrPSc-specific monoclonal antibody we have described previously. RESULTS: The preparation of anti-idiotypic monoclonal antibodies was achieved with well-defined strategies of immunization, selection and subsequent characterization. Our results demonstrate that it is possible to induce a strong anti-idiotypic immune response against the V5B2 monoclonal antibody in both xenogeneic and syngeneic experimental systems. From the competition seen between polyclonal and monoclonal anti-idiotypic antibodies and the original immunogen, the P1 peptide, and even more importantly, the ultimate target antigen, PrPSc, we conclude that selected antibodies bind to the antigen-combining site of the V5B2 monoclonal antibody and might even resemble the PrPSc-specific epitope. The involvement of both antigen-combining sites in the interaction between V5B2 and the most promising monoclonal anti idiotypic antibody was further supported by molecular docking. CONCLUSION: The results of the present study not only provide an example of the successful production of Ab2 monoclonal antibodies based on a well planned strategy for selection, but should also provide a new experimental approach that is applicable to the field of prion diseases. PMID- 19298676 TI - Uncovering new signaling proteins and potential drug targets through the interactome analysis of Mycobacterium tuberculosis. AB - BACKGROUND: Analysis of the pathogen interactome is a powerful approach for dissecting potential signal transduction and virulence pathways. It also offers opportunities for exploring new drug targets. RESULTS: In this study, a protein protein interaction (PPI) network of Mycobacterium tuberculosis H37Rv was constructed using a homogenous protein mapping method, which has shown molecular chaperones, ribosomal proteins and ABC transporters to be highly interconnected proteins. A further analysis of this network unraveled the function of hypothetical proteins as well as a potential signaling pathway. A hypothetical protein, Rv2752c, which was linked to a metal cation-transporting ATPase, was characterized as a metal-beta-lactamase, through domain analysis in combination with an in vitro activity experiment. A second hypothetical protein, Rv1354c, and an unknown protein kinase, PknK, interacted with a similar group of inner membrane-associated ABC transporters in the PPI network. The interactions of Rv1354 with these proteins were also confirmed by a further bacterial two-hybrid analysis. According to protein domain structures, the unique M. tuberculosis Rv1354c gene was proposed, for the first time, to be responsible for the turnover of cyclic-di-GMP, a second messenger molecule in this bacterium. A further structure-based inhibitors screening for Rv1354c was also performed in silicon. CONCLUSION: We constructed a comprehensive protein-protein interaction network for M. tuberculosis consisting of 738 proteins and 5639 interaction pairs. Our analysis unraveled the function of hypothetical proteins as well as a potential signaling pathway. The group of ABC transporters, PknK, and Rv1354c were proposed to constitute a potential membrane-associated signaling pathway that cooperatively responds to environmental stresses in M. tuberculosis. The study therefore provides valuable clues in exploring new signaling proteins, virulence pathways, and drug targets. PMID- 19298677 TI - A new dawn for the use of traditional Chinese medicine in cancer therapy. AB - Although traditional Chinese medicine has benefitted one fifth of the world's population in treating a plethora of diseases, its acceptance as a real therapeutic option by the West is only now emerging. In light of a new wave of recognition being given to traditional Chinese medicine by health professionals and regulatory bodies in the West, an understanding of their molecular basis and highlighting potential future applications of a proven group of traditional Chinese medicine in the treatment of a variety of cancers is crucial - this is where their calling holds much hope and promise in both animal and human trials. Furthermore, the rationale for combining conventional agents and modern biotechnological approaches to the delivery of traditional Chinese medicine is an avenue set to revolutionize the future practice of cancer medicine - and this may well bring on a new dawn of therapeutic strategies where East truly meets West. PMID- 19298678 TI - Measuring the activity of BioBrick promoters using an in vivo reference standard. AB - BACKGROUND: The engineering of many-component, synthetic biological systems is being made easier by the development of collections of reusable, standard biological parts. However, the complexity of biology makes it difficult to predict the extent to which such efforts will succeed. As a first practical example, the Registry of Standard Biological Parts started at MIT now maintains and distributes thousands of BioBrick standard biological parts. However, BioBrick parts are only standardized in terms of how individual parts are physically assembled into multi-component systems, and most parts remain uncharacterized. Standardized tools, techniques, and units of measurement are needed to facilitate the characterization and reuse of parts by independent researchers across many laboratories. RESULTS: We found that the absolute activity of BioBrick promoters varies across experimental conditions and measurement instruments. We choose one promoter (BBa_J23101) to serve as an in vivo reference standard for promoter activity. We demonstrated that, by measuring the activity of promoters relative to BBa_J23101, we could reduce variation in reported promoter activity due to differences in test conditions and measurement instruments by approximately 50%. We defined a Relative Promoter Unit (RPU) in order to report promoter characterization data in compatible units and developed a measurement kit so that researchers might more easily adopt RPU as a standard unit for reporting promoter activity. We distributed a set of test promoters to multiple labs and found good agreement in the reported relative activities of promoters so measured. We also characterized the relative activities of a reference collection of BioBrick promoters in order to further support adoption of RPU-based measurement standards. CONCLUSION: Relative activity measurements based on an in vivoreference standard enables improved measurement of promoter activity given variation in measurement conditions and instruments. These improvements are sufficient to begin to support the measurement of promoter activities across many laboratories. Additional in vivo reference standards for other types of biological functions would seem likely to have similar utility, and could thus improve research on the design, production, and reuse of standard biological parts. PMID- 19298679 TI - The variability and predictors of quality of AIDS care services in Brazil. AB - BACKGROUND: Since establishing universal free access to antiretroviral therapy in 1996, the Brazilian Health System has increased the number of centers providing HIV/AIDS outpatient care from 33 to 540. There had been no formal monitoring of the quality of these services until a survey of 336 AIDS health centers across 7 Brazilian states was undertaken in 2002. Managers of the services were asked to assess their clinics according to parameters of service inputs and service delivery processes. This report analyzes the survey results and identifies predictors of the overall quality of service delivery. METHODS: The survey involved completion of a multiple-choice questionnaire comprising 107 parameters of service inputs and processes of delivering care, with responses assessed according to their likely impact on service quality using a 3-point scale. K means clustering was used to group these services according to their scored responses. Logistic regression analysis was performed to identify predictors of high service quality. RESULTS: The questionnaire was completed by 95.8% (322) of the managers of the sites surveyed. Most sites scored about 50% of the benchmark expectation. K-means clustering analysis identified four quality levels within which services could be grouped: 76 services (24%) were classed as level 1 (best), 53 (16%) as level 2 (medium), 113 (35%) as level 3 (poor), and 80 (25%) as level 4 (very poor). Parameters of service delivery processes were more important than those relating to service inputs for determining the quality classification. Predictors of quality services included larger care sites, specialization for HIV/AIDS, and location within large municipalities. CONCLUSION: The survey demonstrated highly variable levels of HIV/AIDS service quality across the sites. Many sites were found to have deficiencies in the processes of service delivery processes that could benefit from quality improvement initiatives. These findings could have implications for how HIV/AIDS services are planned in Brazil to achieve quality standards, such as for where service sites should be located, their size and staffing requirements. A set of service delivery indicators has been identified that could be used for routine monitoring of HIV/AIDS service delivery for HIV/AIDS in Brazil (and potentially in other similar settings). PMID- 19298680 TI - Effect of pioglitazone versus insulin glargine on cardiac size, function, and measures of fluid retention in patients with type 2 diabetes. AB - BACKGROUND: Both insulin and thiazolidinediones (TZDs) are effective in the treatment of hyperglycaemia and amelioration of insulin resistance in type 2 diabetes but have side effects including weight gain and fluid retention. The use of TZDs has been further hampered by the risk of adverse cardiovascular events including heart failure. The present study evaluated the effect of pioglitazone or insulin glargine on cardiac function and size as well as on surrogate markers of fluid retention such as weight, haemoglobin and natriuretic peptides. METHODS: Thirty patients with inadequate glycaemic control on metformin and sulfonylurea were randomised to receive add-on therapy with insulin glargine or pioglitazone for 26 weeks. Echocardiographic data and blood samples were collected from the two groups before the start of the treatment and after 26 weeks. Left ventricular end-diastolic and left atrial end-systolic volumes were quantified, weight measured and blood samples analyzed. RESULTS: After 26 weeks of treatment, the changes in HbA1c, weight and haemoglobin were similar between the two groups. HDL increased significantly in the pioglitazone group. While there was an increase in natriuretic peptides in the pioglitazone group (NT-proBNP 11.4 +/- 19.6 to 22.8 +/- 44.0, p = 0.046), the difference between the treatment groups was not significant. Left ventricular end-diastolic volume increased by 11% and left atrial end-systolic volume by 17% in the pioglitazone group (Both, p < 0.05, between treatment groups). There was a borderline significant increase in ejection fraction in the pioglitazone group. CONCLUSION: This randomised pilot study showed that six-month treatment with pioglitazone induced significant increases in natriuretic peptides and alterations of cardiac size. These changes were not observed with insulin glargine, which also is known to induce fluid retention. Larger randomised trials are warranted to confirm these findings. PMID- 19298681 TI - Knowledge and attitude on maternal health care among rural-to-urban migrant women in Shanghai, China. AB - BACKGROUND: In China, with the urbanization, women migrated from rural to big cities presented much higher maternal mortality rates than local residents. Health knowledge is one of the key factors enabling women to be aware of their rights and health status in order to seek appropriate health services. This study aims to assess the knowledge and attitude on maternal health care and the contributing factors to being knowledgeable among rural-to-urban migrant women in Shanghai. METHODS: A cross-sectional study was conducted in a district center hospital in Shanghai where migrants gathered. Totally 475 rural-to-urban migrant pregnant women were interviewed and completed the self-administered questionnaire after obtaining informed consent. RESULTS: The mean score of knowledge on maternal health care was 8.28 out of 12. However, only 36.6% women had attended the required 5 antenatal checks, and 58.3% of the subjects thought financial constrains being the main reason for not attending antenatal care. It was found that higher level of education (OR = 3.3, 95%CI: 1.8-3.8), husbands' Shanghai residence (OR = 4.0, 95%CI: 1.3-12.1) and better family income (OR = 3.3, 95%CI: 1.4-8.2) were associated with better knowledge. CONCLUSIONS: Rural-to-urban migrant women's unawareness of maternal health service, together with their vulnerable living status, influences their utilization of maternal health care. Tailored maternal health education and accessible services are in demands for this population. PMID- 19298682 TI - Maternal care and birth outcomes among ethnic minority women in Finland. AB - BACKGROUND: Care during pregnancy and labour is of great importance in every culture. Studies show that people of migrant origin have barriers to obtaining accessible and good quality care compared to people in the host society. The aim of this study is to compare the access to and use of maternity services, and their outcomes among ethnic minority women having a singleton birth in Finland. METHODS: The study is based on data from the Finnish Medical Birth Register in 1999-2001 linked with the information of Statistics Finland on woman's country of birth, citizenship and mother tongue. Our study data included 6,532 women of foreign origin (3.9% of all singletons) giving singleton birth in Finland during 1999-2001 (compared to 158,469 Finnish origin singletons). RESULTS: Most women have migrated during the last fifteen years, mainly from Russia, Baltic countries, Somalia and East Europe. Migrant origin women participated substantially in prenatal care. Interventions performed or needed during pregnancy and childbirth varied between ethnic groups. Women of African and Somali origin had most health problems resulted in the highest perinatal mortality rates. Women from East Europe, the Middle East, North Africa and Somalia had a significant risk of low birth weight and small for gestational age newborns. Most premature newborns were found among women from the Middle East, North Africa and South Asia. Primiparous women from Africa, Somalia and Latin America and Caribbean had most caesarean sections while newborns of Latin American origin had more interventions after birth. CONCLUSION: Despite good general coverage of maternal care among migrant origin women, there were clear variations in the type of treatment given to them or needed by them. African origin women had the most health problems during pregnancy and childbirth and the worst perinatal outcomes indicating the urgent need of targeted preventive and special care. These study results do not confirm either healthy migrant effect or epidemiological paradox according to which migrant origin women have considerable good birth outcomes. PMID- 19298683 TI - Influence of fruit drinks with or without lactobacillus Lp299v on the gastrointestinal uptake of paracetamol in man. AB - BACKGROUND: Clinical observations have revealed that patients throw up undigested paracetamol tablets several hours following intake of rosehip drink with Lp299v (Proviva). The purpose of this study was to demonstrate if this translates into altered plasma levels of paracetamol in nineteen healthy subjects consuming 200 ml of water, Rose hip drink or Proviva together with 1.5 gram of paracetamol. FINDINGS: The concentration of paracetamol in plasma increased rapidly when the paracetamol-containing tablets were consumed together with water and after 30 minutes a median level of 90 mumol/l was reached (a 95% confidence interval of 57,161). The corresponding 30 minutes values of paracetamol levels in the presence of rosehip with or without Lp299v were 0 mumol/l (95% confidence intervals contain only zero for both rosehip treatments). There were significant differences in AUC, maximal paracetamol concentration and in time to maximal paracetamol concentration. The median maximal paracetamol concentration was 147 mumol/l for water, which is significantly higher than the median for rosehip drink with Lp299v,113.5 mumol/l, and than the median for rosehip-drink without Lp299v, 106.5 mumol/l (p = 0.002, and p = 0.003); there were no significant difference between rosehip drink with or without Lp299v (p > 0.3). CONCLUSION: We have demonstrated an interaction between the uptake of paracetamol and the solvent in rosehip drink/Provia which mainly consists of long chain carbohydrates. This may in the clinic translate to the use of more drug than it is necessary. PMID- 19298684 TI - Trans-10, cis-12-conjugated linoleic acid reduces the hepatic triacylglycerol content and the leptin mRNA level in adipose tissue in obese Zucker fa/fa rats. AB - Conjugated linoleic acid (CLA) isomers have been reported to reduce body weight and beneficially affect glucose metabolism in animals, but the results are inconsistent and seem to depend on animal model and type of CLA isomer. In the present study, feeding male Zucker fa/fa rats diets supplemented with 1% trans 10, cis-12-CLA for 10 d reduced the liver TAG content without improving the overall adiposity, and enhanced hepatic mitochondrial and peroxisomal beta oxidation. The increased carnitine palmitoyltransferase (CPT)-I activity and mRNA level as well as the increased n-3:n-6 PUFA ratio in liver suggest that trans-10, cis-12-CLA increased the hepatic beta-oxidation by stimulation of PPARalpha. The reduced hepatic TAG content may be partly due to lower activity of stearoyl-CoA desaturase, as the ratios of 18 : 1n-9:18 : 0 and 16 : 1n-7:16 : 0 were reduced in liver. Trans-10, cis-12-CLA increased the CPT-I mRNA in retroperitoneal white adipose tissue (WAT), and increased uncoupling protein-2 mRNA in epididymal and inguinal WAT depots. Leptin mRNA level was decreased in all examined WAT depots, implying reduced insulin sensitivity. The resistin mRNA level was increased in all WAT depots, whereas adiponectin mRNA was reduced in inguinal and retroperitoneal WAT. The present results suggest that dietary supplementation with trans-10, cis-12-CLA may increase the catabolism of lipids in liver and adipose tissue. Moreover, we provide new data suggesting that trans-10, cis-12 CLA modulates the expression of resistin and adiponectin inversely in adipose tissue. Hence, the present results suggest that trans-10, cis-12-CLA may have some beneficial effects on lipid metabolism and adiposity but possibly reduces insulin sensitivity. PMID- 19298685 TI - Stereotype threat contributes to social difficulties in people with schizophrenia. AB - OBJECTIVE: The experience of stereotype threat (where the prospect of conforming to a stereotype, or of being treated in terms of it, becomes self-threatening) affects members of social groups about whom devaluing stereotypes exist. Although a widely endorsed stereotype of schizophrenia concerns social skill impairment, it is unclear whether the experience of stereotype threat impacts social functioning in this group. The purpose of the present study was to test whether people with schizophrenia would perform more poorly in a social setting in which they felt stereotyped as mentally ill. METHODS: Thirty individuals with schizophrenia engaged in conversations with two confederates, one of whom they were told knew nothing about them (control conversation), and the other of whom they were told had been informed of their diagnosis (stereotype threat conversation). In reality, neither confederate had been informed of participants' mental health status. RESULTS: Although participants with schizophrenia did not perceive any differences in their own social behaviour across the two conditions, their social skill was rated by the confederates as poorer in the stereotype threat conversation on three out of the six measures used. CONCLUSIONS: These results suggest that social skill difficulties in people with schizophrenia may be exacerbated by their awareness that others know of their diagnosis. These findings have implications for disclosure of mental health status. PMID- 19298686 TI - The sacred lotus (Nelumbo nucifera) - phytochemical and therapeutic profile. AB - OBJECTIVES: Nelumbo nucifera Gaertn. (Nymphaeaceae), also known as sacred lotus, is a well known medicinal plant. This article reviews the traditional uses, phytochemistry and therapeutic reports on different parts of N. nucifera viz. the seeds, rhizomes, leaves and flowers. This review also describes various compounds isolated from different parts of this plant and the therapeutic benefits derived from those phytoconstituents. KEY FINDINGS: There are several therapeutic benefits of this plant for which different parts are used. The extracts of rhizomes, seeds, flowers and leaves have been reported to have varied therapeutic potential. Several bioactive compounds have been derived from these plant parts belonging to different chemical groups, including alkaloids, flavonoids, glycosides, triterpenoid, vitamins etc., which all have their own therapeutic impact. Thus, the pharmacological effects and various active ingredients of different parts of N. nucifera are well understood. SUMMARY: In this review we explore the current pharmaceutical, phytochemical and pharmacological knowledge about this well known plant species as well as several emerging aspects for research on N. nucifera. PMID- 19298687 TI - Phycocyanin liposomes for topical anti-inflammatory activity: in-vitro in-vivo studies. AB - OBJECTIVES: The aim of this work was to investigate the anti-inflammatory activity of C-phycocyanin (C-PC) on skin inflammation after topical administration and the influence of liposomal delivery on its pharmacokinetic properties. METHODS: Liposomes of different size and structure were prepared with different techniques using soy phosphatidylcholine and cholesterol. Vesicular dispersions were characterised by transmission electron microscopy, optical and fluorescence microscopy for vesicle formation and morphology, dynamic laser light scattering for size distribution, and Zetasizer for zeta-potential. C-PC skin penetration and permeation experiments were performed in vitro using vertical diffusion Franz cells and human skin treated with either free or liposomal drug dispersed in a Carbopol gel. KEY FINDINGS: The protein was mainly localised in the stratum corneum, while no permeation of C-PC through the whole skin thickness was detected. Two percent C-PC-encapsulating liposomes showed the best drug accumulation in the stratum corneum and the whole skin, higher than that of the corresponding free 2% C-PC gel. Moreover, skin deposition of liposomal C-PC was dose dependent since skin accumulation values increased as the C-PC concentration in liposomes increased. The topical anti-inflammatory activity of samples was evaluated in vivo as inhibition of croton oil-induced or arachidonic acid-induced ear oedema in rats. CONCLUSIONS: The results showed that C-PC can be successfully used as an anti-inflammatory drug and that liposomal encapsulation is effective in improving its anti-inflammatory activity. PMID- 19298688 TI - A study on the effect of inorganic salts in transungual drug delivery of terbinafine. AB - OBJECTIVES: The poor success rate of topical therapy in nail disorders is mainly because of the low permeability of keratinized nail plates. This can be overcome by utilizing potent perungual drug penetration enhancers that facilitate the drug permeation across the nail plate. This study evaluated the efficacy of inorganic salts in enhancing the trans-nail permeation using a model potent antifungal agent, terbinafine hydrochloride. METHODS: Permeation studies were carried out across human cadaver nail in a Franz diffusion cell using terbinafine solution (1 mg/ml; pH 3). Preliminary studies were carried out to assess the effect of salts (0.5 M) on the terbinafine permeation into and through the nail. Further, the influence of salt concentration (0.25-3 M) on permeation, the mechanism for the enhancement and the suitability of developing a formulation were also studied. KEY FINDINGS: Terbinafine permeation (3-5 fold) through the nail and drug load (4 7 fold) in the nail were enhanced significantly when salts were used at 0.5 M concentration. Increase in salt concentration up to 1 M increased the permeation, which decreased with further increase in salt concentration (>1 M). Mechanistic studies revealed that the enhanced permeation by salts was mainly due to their ability to increase the nail hydration and also to increase the thermodynamic activity of the drug. The cumulative amount of terbinafine permeated at 24 h from the formulated gel (9.70 +/- 0.93 microg/cm(2)) was comparable with that of a solution (11.45 +/- 1.62 microg/cm(2)). CONCLUSIONS: Given the promising results from the permeation and drug load studies, it was concluded that inorganic salts could be used as potent transungual permeation enhancers. PMID- 19298689 TI - Effects of concurrent administration of nevirapine on the disposition of quinine in healthy volunteers. AB - OBJECTIVES: Nevirapine and quinine are likely to be administered concurrently in the treatment of patients with HIV and malaria. Both drugs are metabolised to a significant extent by cytochrome P450 (CYP)3A4 and nevirapine is also an inducer of this enzyme. This study therefore evaluated the effect of nevirapine on the pharmacokinetics of quinine. METHODS: Quinine (600 mg single dose) was administered either alone or with the 17th dose of nevirapine (200 mg every 12 h for 12 days) to 14 healthy volunteers in a crossover fashion. Blood samples collected at predetermined time intervals were analysed for quinine and its major metabolite, 3-hydroxquinine, using a validated HPLC method. KEY FINDINGS: Administration of quinine plus nevirapine resulted in significant decreases (P < 0.01) in the total area under the concentration-time curve (AUC(T)), maximum plasma concentration (C(max)) and terminal elimination half-life (T((1/2)beta)) of quinine compared with values with quinine dosing alone (AUC: 53.29 +/- 4.01 vs 35.48 +/- 2.01 h mg/l; C(max): 2.83 +/- 0.16 vs 1.81 +/- 0.06 mg/l; T((1/2)beta): 11.35 +/- 0.72 vs 8.54 +/- 0.76 h), while the oral plasma clearance markedly increased (11.32 +/- 0.84 vs 16.97 +/- 0.98 l/h). In the presence of nevirapine there was a pronounced increase in the ratio of AUC(metabolite)/AUC (unchanged drug) and highly significant increases in C(max) and AUC of the metabolite (P < 0.01). CONCLUSIONS: Nevirapine significantly alters the pharmacokinetics of quinine. An increase in the dose of quinine may be necessary when the drug is co administered with nevirapine. PMID- 19298690 TI - Oral bioavailability enhancement of a hydrophilic drug delivered via folic acid coupled liposomes in rats. AB - OBJECTIVES: A liposome preparation that is amenable to receptor-mediated endocytosis has been developed to enhance the oral bioavailability of poorly absorbable peptidomimetic drugs by use of folic acid as the mediator of liposomal uptake. METHODS: Folic acid was physically coupled to the surface of the liposomes and cefotaxime was used as the model drug. In-vivo evaluation was carried out on eight Sprague-Dawley rats in a two-way crossover study to compare the oral bioavailability of cefotaxime loaded in folic acid-free liposomes and folic acid-coupled liposomes. Blood samples were collected from the tail vein and plasma cefotaxime levels were determined using an HPLC method. KEY FINDINGS: Enhanced oral bioavailability (AUC(0-infinity)) of cefotaxime was observed when administered via folic acid-coupled liposomes. The peak plasma concentration (C(max)) of cefotaxime was increased when administered via folic acid-coupled liposomes as compared with folic acid-free liposomes. At 90% confidence interval, the value for AUC(0-infinity) was 1.4-2-times higher and the value for C(max) was 1.2-1.8-times higher for the folic acid-coupled liposomes compared with folic acid-free liposomes. CONCLUSIONS: Folic acid could enhance the uptake of liposomally entrapped drug. It could be a useful candidate to supplement liposome delivery systems. PMID- 19298691 TI - Effect of rutin on warfarin anticoagulation and pharmacokinetics of warfarin enantiomers in rats. AB - OBJECTIVES: The effects of the flavonoid rutin on the anticoagulant activity of oral warfarin and the protein binding and pharmacokinetics of its enantiomers were investigated in rats. METHODS: A single dose of racemic warfarin, 1.5 mg/kg, was administered orally to rats either alone or on day 5 of an 8-day oral regimen of rutin, 1 g/kg daily. RESULTS: Rutin reduced the anticoagulant effect of racemic warfarin, evident as a 31% reduction in the area under the prothrombin complex activty-time curve (P < 0.05). KEY FINDINGS: Rutin had no apparent effect on pre-treatment baseline blood coagulation. It enhanced the in-vitro serum protein binding of S- and R-warfarin (reflected by 40% and 26% reductions in unbound fraction, respectively), and thus restricted distribution by 33 and 21%, respectively. Treatment with rutin significantly decreased the elimination half life of S-warfarin by 37% as a result of the 69% increase in unbound clearance of the S-enantiomer. This effect was attributed to a significant 77% increase in the unbound formation clearance of the overall oxidative and reductive metabolites, and an increase in the unbound renal clearance of the more potent S-enantiomer of warfarin. CONCLUSIONS: Concurrent rutin administration is likely to reduce the anticoagulant effect of racemic warfarin, reflecting a significant decrease in the elimination half-life of the more potent S-enantiomer. PMID- 19298692 TI - Montmorillonite adsorbs creatinine and accelerates creatinine excretion from the intestine. AB - OBJECTIVES: This study aims to evaluate the sorption by montmorillonite of creatinine and the accelerating effect of montmorillonite on creatinine excretion from the intestine. METHODS: The sorption of montmorillonite was observed in vitro. Also, rat intestinal tract and blood vessels were perfused circularly with perfusate with or without creatinine, respectively, to study the promotion of creatinine diffusion from the blood vessel to the intestine and the inhibition of creatinine absorption in the intestinal tract. The effect of decreasing the serum concentration of creatinine was studied in an acute hypercreatininaemia mouse model. The concentration of creatinine was determined by the basic picric acid method. KEY FINDINGS: Montmorillonite adsorbed creatinine markedly in the simulated intestinal solution in a concentration-dependent manner. The sorption time curve of montmorillonite with creatinine showed that the sorption was fast. The adsorption rate reached a maximum in 10 min. The pH of the solution influenced the sorption, the rate of which was higher at a low pH than at a high pH. Creatinine could diffuse from the blood vessel to the intestine and was reabsorbed in the intestine. Montmorillonite promoted the diffusion and inhibited the absorption. Montmorillonite decreased the serum creatinine level of hypercreatininaemia mice prepared by injecting creatinine intraperitoneally. CONCLUSIONS: Montmorillonite adsorbs creatinine and accelerates its excretion from the intestine. PMID- 19298693 TI - Discrepancies between predicted and observed rates of intravenous gentamicin delivery for neonates. AB - OBJECTIVES: This study aimed to investigate intravenous infusions as used in the neonatal intensive care setting, to determine the effect of gentamicin dose (mg), gentamicin concentrations (mg/ml), flow rate (ml/h) and flush volume (ml) upon the length of infusion time. METHODS: Intravenous infusions were set up to simulate administration of gentamicin to neonates. Dextrose (10%, w/v) was administered as the primary intravenous fluid at 3.8 or 18.7 ml/h. Gentamicin doses (0.5 mg/0.2 ml, 2 mg/0.2 ml, 2.5 mg/1.0 ml, or 10 mg/1.0 ml) were delivered into the intravenous line at a T-connection using a Graseby pump over 35 min. This was followed by a saline flush of 1 or 2 ml over a further 35 min. At the end of each experiment a 2 ml 0.9% saline bolus was given. Analysis of gentamicin collected at 5-min intervals was by an HPLC method. KEY FINDINGS: The experiment demonstrated that under the infusion conditions neonates weighing 2.5 kg would receive only 80% of the drug at 60 min, increasing to 90-95% by 75 min. In extremely low birth weight neonates (0.5 kg), even lower percentage of gentamicin recovery occurred. At 60 min only 60% of the intended gentamicin dose had been delivered and this increased to only 70% by 75 min. CONCLUSIONS: The delivery of gentamicin administered by intravenous infusion is substantially extended in extremely low birth weight neonates. This appeared to be primarily due to the small volumes and low infusion rates used in these patients. PMID- 19298694 TI - Protective effects of coffee-derived compounds on lipopolysaccharide/D galactosamine induced acute liver injury in rats. AB - OBJECTIVES: The protective effects of coffee-derived compounds on lipopolysaccharide/D-galactosamine (LPS/D-GalN) induced acute liver injury in rats were investigated. METHODS: Wistar rats were orally administered saline (control) or one of the test compounds (caffeine, chlorogenic acid, trigonelline, nicotinic acid or eight pyrazinoic acids) at a dose of 100 mg/kg, respectively. This was followed by intraperitoneal injection with LPS (100 mug/kg)/D-GalN (250 mg/kg) 1 h after administration of the test compounds. Blood samples were collected up to 12 h after LPS/D-GalN injection, followed by determination of plasma aspartate aminotransferase, alanine aminotransferase, tumour necrosis factor alpha (TNF-alpha) and interleukin 10 (IL-10) levels. KEY FINDINGS: Plasma aspartate aminotransferase and alanine aminotransferase levels were significantly increased after LPS/D-GalN-treatment, but were suppressed by pretreatment with caffeine (n = 5), nicotinic acid, non-substituted pyrazinoic acid or 5 methylpyrazinoic acid (n = 6, respectively) 12 h after LPS/D-GalN-treatment (P < 0.01, respectively). Moreover, the animals pretreated with these test compounds showed significantly higher survival rates (83-100%) compared with the control (23%). Only pretreatment with caffeine significantly suppressed the LPS/D-GalN induced elevation of plasma TNF-alpha levels 1 and 2 h after LPS/D-GalN-treatment (P < 0.01, respectively). Pretreatment with caffeine, nicotinic acid or non substituted pyrazinoic acid activated the LPS/D-GalN induced elevation of plasma IL-10 levels at 1 and 2 h, although there were no statistically significant differences in IL-10 levels between control and nicotinic acid or non-substituted pyrazinoic acid treated rats. CONCLUSIONS: The results suggest that caffeine, nicotinic acid, non-substituted pyrazinoic acid and 5-methylpyrazinoic acid can protect against LPS/D-GalN induced acute liver injury, which may be mediated by the reduction of TNF-alpha production and/or increasing IL-10 production. PMID- 19298695 TI - Bis-(3-hydroxyphenyl) diselenide inhibits LPS-stimulated iNOS and COX-2 expression in RAW 264.7 macrophage cells through the NF-kappaB inactivation. AB - OBJECTIVES: Previously, we reported that diaryl diselenide compounds have strong inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in macrophages. In this study, we investigated the molecular mechanisms underlying NO suppression and prostaglandin E(2) (PGE(2)) production by diaryl diselenide compounds, bis-(2-hydroxyphenyl) diselenide (DSE-A), bis-(3 hydroxyphenyl) diselenide (DSE-B), bis-(4-hydroxyphenyl) diselenide (DSE-C), dipyridyl diselenide (DSE-D) and diphenyl diselenide (DSE-E). METHODS: The effect of these compounds on NO suppression and PGE(2) production was investigated in RAW 264.7 macrophages. KEY FINDINGS: Our data indicate that of the above, DSE-B most potently inhibits NO and PGE(2) production, and that it also significantly reduces the releases of tumour necrosis factor (TNF)-alpha, interleukin(IL)-1beta and IL-6. Consistent with these observations, DSE-B also reduced the protein levels of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), and the mRNA levels of iNOS, COX-2, TNF-alpha, IL-1beta and IL-6. Furthermore, DSE-B inhibited LPS-induced nuclear factor-kappaB (NF-kappaB) activation, which was associated with the prevention of the inhibitor kappaB-alpha (IkappaB-alpha) degradation and a subsequent reduction in nuclear p65 protein levels. CONCLUSIONS: Taken together, our data suggest that the anti-inflammatory properties of DSE-B are due to reduction in the expression of iNOS, COX-2, TNF-alpha, IL-1beta and IL-6 through the down-regulation of NF-kappaB binding activity. PMID- 19298696 TI - Morphine or its withdrawal affects plasma malondialdehyde, vitamin E levels and absence or presence of abstinence signs in rats. AB - OBJECTIVES: Various experimental observations show that morphine treatment generates reactive oxygen species, and that its discontinuation leads to signs of withdrawal. We therefore investigated plasma malondialdehyde and vitamin E levels under both conditions to verify the occurrence of any alterations in oxidative metabolism, and whether these are associated with behavioural changes. METHODS: We investigated the effects of morphine or morphine plus naloxone on plasma malondialdehyde, vitamin E levels and withdrawal signs such as jumping, wet dog shakes and faecal excretion in rats. Furthermore, isopropylnoradrenaline was injected in rabbits to verify its effects on plasma malondialdehyde levels. KEY FINDINGS: Morphine treatment increased free malondialdehyde and decreased vitamin E levels. The elevation in malondialdehyde levels were exacerbated by the abrupt removal of morphine by naloxone, which also led to the appearance of withdrawal signs. The increased malondialdehyde values can be attributed to the interactions of reactive oxygen species with unsaturated fatty acids, and the lowered levels of vitamin E to its interactions with reactive oxygen species. CONCLUSIONS: A connection seems to exist between altered peroxide status and withdrawal signs in abstinent animals. PMID- 19298697 TI - Effects of co-administration of bupropion and nicotine or D-amphetamine on the elevated plus maze test in mice. AB - OBJECTIVES: A variety of abused drugs, including psychostimulants, can modulate the expression of anxiety. Although the effect of nicotine and D-amphetamine on anxiety-related behaviour in animal models has been investigated, the mechanisms underlying the anxiogenic or anxiolytic actions of these drugs have not been clarified. Bupropion is an antidepressant drug which may alleviate some symptoms of nicotine withdrawal, although its effects on anxiety are not clear. We have investigated the effect of nicotine and D-amphetamine on anxiety in the elevated plus maze test in mice. METHODS: We examined the influence of acute administration of nicotine (0.1 mg/kg, s.c.) and D-amphetamine (2 mg/kg, i.p.) on anxiety level. We then evaluated the anxiety-related response after subchronic injection of both psychostimulants, including crossover effects. For this purpose, nicotine (0.1 mg/kg, s.c.) was administered daily for six days, and on the seventh day mice were challenged with nicotine (0.1 mg/kg, s.c.) or D amphetamine (2 mg/kg, i.p.). A distinct group of mice was pretreated with D amphetamine (2 mg/kg, i.p., 8 days), and subjected to D-amphetamine (2 mg/kg, i.p.) or nicotine (0.1 mg/kg, s.c.) challenge on the ninth day. Moreover, we investigated acute and subchronic effects of coadministration of bupropion (5, 10 and 20 mg/kg; i.p.) and nicotine or D-amphetamine. KEY FINDINGS: We observed that acute anxiogenic effects of nicotine and D-amphetamine as well as the development of tolerance and cross-tolerance to their effects were blunted by a pretreatment with a nonactive dose of bupropion (5 mg/kg, i.p.). CONCLUSIONS: These results demonstrated that similar neural mechanisms were involved in the regulation of nicotine and D-amphetamine anxiety-like behaviour in mice. The results have provided new findings to support the use of bupropion in the treatment of nicotine and/or amphetamine addiction. PMID- 19298698 TI - Synergism of simvastatin with losartan prevents angiotensin II-induced cardiomyocyte apoptosis in vitro. AB - OBJECTIVES: Increasing evidence suggests that cardiomyocyte apoptosis has an important role in the transition from compensatory cardiac remodelling to heart failure. The synergistic effect of statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors) and angiotensin II (Ang II) type 1 receptor antagonists reduces the incidence of cardiovascular events. However, the anti-apoptotic potential of the synergism between losartan and simvastatin in heart failure remains unexplored. Here, we demonstrate that Ang II-induced apoptosis is prevented by losartan and simvastatin in neonatal cardiomyocytes. METHODS: The in vitro cardiomyocyte apoptosis model was established by co-culturing neonate rat cardiomyocytes with Ang II. Cell viability was analysed by the MTT assay. Cell apoptosis was evaluated using fluorescence microscopy and flow cytometry. Apoptosis-related proteins Bax and Bcl-2 expressions were measured by flow cytometry detection. KEY FINDINGS: Incubation with 10(-7) M Ang II for 48 h increased cardiomyocyte apoptosis and decreased cell viability. Losartan (10(-5) M) and simvastatin (10(-5) M), either alone or in combination, significantly decreased Ang II-induced cardiomyocyte apoptosis and increased cell viability. The q values calculated by the probability sum test were 1.31 for cardiomyocyte apoptosis and 1.21 for cell viability. Ang II induced a significant increase in Bax protein expression, whereas Bcl-2 protein expression was decreased. Losartan alone or in combination with simvastatin blocked the increased Bax expression and increased Bcl-2 expression. However, simvastatin had no such effect. CONCLUSIONS: Our data provide the first evidence that synergism of simvastatin with losartan prevents angiotensin II-induced cardiomyocyte apoptosis in vitro. Synergism between simvastatin and losartan may provide a new therapeutic approach to the prevention of cardiac remodelling. PMID- 19298699 TI - Antiplatelet effects of piplartine, an alkamide isolated from Piper tuberculatum: possible involvement of cyclooxygenase blockade and antioxidant activity. AB - OBJECTIVES: Piplartine (piperlongumine; 5,6-dihydro-1-[1-oxo-3-(3,4,5 trimethoxyphenyl]-2(1H) pyridinone) is an alkaloid amide isolated from Piper species (Piperaceae). It has been reported to show multiple pharmacological activities in vitro and in vivo. METHODS: We evaluated the in-vitro antiplatelet effect of piplartine isolated from the roots of P. tuberculatum, on human platelet aggregation induced in platelet-rich plasma by the agonists collagen, adenosine 5'-diphosphate (ADP), arachidonic acid (AA) and thrombin. KEY FINDINGS: Piplartine (100 mug/ml) caused a 30% inhibition in platelet aggregation when collagen was the agonist. At 200 mug/ml, piplartine significantly inhibited the aggregation induced by arachidonic acid (100%), collagen (59%) or ADP (52%) but not that induced by thrombin. The highest concentration of piplartine (300 mug/ml) inhibited thrombin- (37%), ADP- (71%) and collagen- (98%) induced aggregation. The inhibitory effect of piplartine on ADP-induced platelet aggregation was not modified by pretreatment with pentoxifylline (a phosphodiesterase inhibitor), L-arginine (a substrate for nitric oxide synthase) or ticlopidine (a P2Y(12) purinoceptor antagonist). However, aspirin, a well known inhibitor of cyclooxygenase, greatly increased the inhibitory effect of piplartine on arachidonic-acid-induced platelet aggregation. CONCLUSIONS: The mechanism underlying the piplartine antiplatelet action is not totally clarified. It could be related to the inhibition of cyclooxgenase activity and a decrease in thromboxane A(2) formation, similar to that occurring with aspirin. This and other possible mechanisms require further study. PMID- 19298700 TI - Eugenosedin-A prevents hyperglycaemia, hyperlipidaemia and lipid peroxidation in C57BL/6J mice fed a high-fat diet. AB - OBJECTIVES: Eugenosedin-A is a serotonin (5-hydroxytryptamine; 5-HT) 5-HT(1B/2A) and alpha(1)/alpha(2)/beta(1)-adrenoceptor blocker with anti-oxidative, anti inflammatory and free-radical scavenging activities. Previous reports demonstrated that 5-HT(2A) blockers could diminish hyperlipidaemia. This study therefore aimed to investigate the possible uses and mechanisms of eugenosedin-A and other agents in treating hyperlipidaemia. METHODS: C57BL/6J mice were randomly divided into seven groups, fed a regular diet or a high-fat diet alone or supplemented with one of five agents: eugenosedin-A, ketanserin, prazosin, propranolol or atorvastatin (5 mg/kg p.o.) for 8 weeks. KEY FINDINGS: Compared with the regular diet, the mice fed the high-fat diet had significantly higher body weight and glucose, insulin and lipid levels. Brain malondialdehyde concentration was increased and liver glutathione peroxidase activity decreased. Addition of eugenosedin-A to the high-fat diet resulted in less weight gain and reduced hyperglycaemia, hyperinsulinaemia and hyperlipidaemia. Lipid and glucose homeostasis were related to decreased hepatic lipogenesis mRNAs and proteins (sterol regulatory element binding protein 1a, fatty acid synthase, sterol-CoA desaturase) and restored adipose peroxisome proliferator-activated receptor gamma expression. Eugenosedin-A also enhanced low-density lipoprotein receptor mRNA expression. CONCLUSIONS: Eugenosedin-A may improve plasma lipid metabolism by increasing low-density lipoprotein receptor and peroxisome proliferator-activated receptor gamma expression and diminishing sterol regulatory element binding protein 1a, fatty acid synthase and sterol-CoA desaturase. Reduction of plasma glucose and lipid levels may, in turn, reduce insulin concentration, which would explain the marked improvement in obesity-related hyperglycaemia and hyperlipidaemia. Furthermore, eugenosedin-A affected malondialdehyde concentration and glutathione peroxidase activity, suggesting it may have anti peroxidation effects in mice fed a high-fat diet. PMID- 19298701 TI - Effects of anticancer agents and scavengers on CMV-promoter-driven exogenous gene expression in genetically modified cells. AB - OBJECTIVES: This study aimed to investigate whether the levels of rsGFP mRNA and the fluorescence levels of cytomegalovirus (CMV)-promoter-driven rsGFP (red shifted green fluorescent protein) could be changed by using anticancer agents and also to examine the effects of co-treatment with anticancer agents and scavengers. METHODS: The pQBI25 vector, which encodes the CMV promoter and the cDNA for rsGFP, was transfected into FR cells (rat skin fibroblast cell line). FR pQBI25 cells were then exposed to doxorubicin, 5-fluorouracil, methotrexate or paraquat with or without scavengers such as N-acetyl cysteine (NAC) and edaravone for 48 h. KEY FINDINGS: The levels of rsGFP mRNA were found to be significantly higher following doxorubicin, 5-fluorouracil and paraquat treatment but were not changed by methotrexate. These levels of rsGFP mRNA were found to be significantly lower after paraquat/edaravone co-treatment compared with paraquat alone. The fluorescence levels of rsGFP were found to be significantly higher following doxorubicin and paraquat treatment but were not changed by 5 fluorouracil and methotrexate. The levels were also found to be significantly lower after paraquat/edaravone co-treatment compared with paraquat alone and also after doxorubicin/NAC co-treatment compared with doxorubicin alone. CONCLUSIONS: These findings suggest that CMV-promoter-driven exogenous gene expression may be partly regulated by reactive oxygen species. PMID- 19298702 TI - Reichardt's dye and its reactions with the alkylating agents 4-chloro-1-butanol, ethyl methanesulfonate, 1-bromobutane and Fast Red B - a potentially useful reagent for the detection of genotoxic impurities in pharmaceuticals. AB - OBJECTIVES: Alkylating agents are potentially genotoxic impurities that may be present in drug products. These impurities occur in pharmaceuticals as by products from the synthetic steps involved in drug production, as impurities in starting materials or from in-situ reactions that take place in the final drug product. Currently, analysis for genotoxic impurities is typically carried out using either HPLC/MS or GC/MS. These techniques require specialist expertise, have long analysis times and often use sample clean-up procedures. Reichardt's dye is well known for its solvatochromic properties. In this paper the dye's ability to undergo alkylation is reported. METHODS: The reaction between Reichardt's dye and alkylating agents such as 4-chloro-1-butanol and ethyl methanesulfonate was monitored spectrophotometrically at 618 nm in acetonitrile and 624 nm in N,N-dimethylformamide. KEY FINDINGS: Changes in absorption were observed using low levels of alkylating agent (5-10 parts per million). Alkylation of the dye with 4-chloro-1-butanol and ethyl methanesulfonate was confirmed. Reichardt's dye, and its changing UV absorption, was examined in the presence of paracetamol (10 and 100 mg/ml). Whilst the alkylation-induced changes in UV absorption were not as pronounced as with standard solutions, detection of alkylation was still possible. CONCLUSIONS: Using standard solutions and in the presence of a drug matrix, Reichardt's dye shows promise as a reagent for detection of low levels of industrially important alkylating agents. PMID- 19298703 TI - Validation of LC for the determination of alpha-mangostin in mangosteen peel extract: a tool for quality assessment of Garcinia mangostana L. AB - Mangosteen, Garcinia mangostana L., is known as the "Queen of fruits" and can be cultivated in the tropical rainforest such as Malaysia, Indonesia, and Thailand. Compounds isolated from the fruit peel of mangosteen contain abundant xanthones (especially alpha-mangostin). It has been used as traditional medicine such as anti-inflammatory and antibacterial and is popularly applied to cosmetic and pharmaceutical products. However, there is little information for quality and quantity determination of alpha-mangostin in mangosteen. Thus, the aim of this study was to set up a validated and stability-indicated isocratic reverse-phase high-performance liquid chromatographic (HPLC) method for quality control and quantity determination of a-mangostin from mangosteen peel extract. The assay was fully validated and shown to be linear (r(2) > 0.999), sensitive (LOD = 0.02 microg/mL and LOQ = 0.08 microg/mL), accurate (intra-day was between 98.1-100.8%, inter-day was between 90.0-101.3%), precise (intra-day variation < or = 1.8%, inter-day variation < or = 4.3%), specific, and with good recovery. Total analysis was approximately 8 min. The finalized method is also a stability indicating assay. The present method should be useful for analytical research and for routine quality control analysis of alpha-mangostin in mangosteen peel extract and products of mangosteen. PMID- 19298704 TI - Rapid identification of polyphenol C-glycosides from Swertia franchetiana by HPLC ESI-MS-MS. AB - High-performance liquid chromatography coupled to positive ion electrospray ionization tandem mass spectrometry (MS) and diode array detection was employed to identify the polyphenol C-glycosides in the extract of Swertia franchetiana, a traditional Chinese/Tibetan herb. The neutral loss scan of the extract of S. franchetiana using the characteristic losses of 120 and 150 u provided a detailed profile of the polyphenol C-glycosides in the complex mixture. On-line UV spectroscopy along with MS-MS and MS-MS-MS mass spectra analysis produced with and without in-source collision induced dissociation was contributed to discriminate and identify the polyphenol C-glycosides. Three xanthone C glycosides (i.e., mangiferin, isomangiferin, and 1,6,7-trihydroxyl-2-C glucosexanthone) and three flavone C-glycosides (i.e., isoorientin, isovitexin, and swertisin) were tentatively identified. Isomangiferin and 1,6,7-trihydroxyl-2 C-glucosexanthone were for the first time found in this plant. PMID- 19298705 TI - Quantitative HPLC determination and extraction of anthraquinones in Senna alata leaves. AB - A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of four anthraquinones: rhein, aloe-emodin, emodin, and chrysophanol in Senna alata leaves. The method involves the use of a TSK-gel ODS-80Tm column (5 microm, 4.6 x 150 mm) at 25 degrees C with the mixture of methanol and 2% aqueous acetic acid (70:30, v/v) as the mobile phase and detection at 254 nm. The parameters of linearity, precision, accuracy, and specificity of the method were evaluated. The recovery of the method is 100.3 100.5%, and linearity (r(2) > 0.9998) was obtained for all anthraquinones. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 5%) were also achieved. The solvent for extraction of anthraquinones from S. alata leaves was examined in order to increase the anthraquinone content of the leaf extract. It was found that a solution of 5% hydrochloric acid (v/v), 5% ferric chloride (w/v), and 15% water in methanol (v/v) was capable of increasing the anthraquinone content in the leaf extract up to 1.67% (w/w). PMID- 19298706 TI - HPLC determination of eight polyphenols in the leaves of Crataegus pinnatifida Bge. var. major. AB - A simple high-performance liquid chromatographic (HPLC) assay using the internal standard method is developed for the simultaneous determination of eight polyphenols. The analyzed compounds isolated from the leaves of Crataegus pinnatifida Bge. var. major include chlorogenic acid, vitexin-4"-O-glucoside, vitexin-2"-O-rhamnoside, vitexin, rutin, hyperoside, isoquercitrin, and quercetin. HPLC analysis is performed on a Diamonsil C18 analytical column (150 x 4.6 mm, i.d., 5-microm) using solvent (A) acetonitrile-tetrahydrofuran (95:5, v/v) and (B) 1% aqueous phosphoric acid as the mobile phase with UV absorption at 270 nm. The calibration curves of the eight polyphenols are linear (r(2) > 0.9992) over the concentration range of 0.0894-120.0 microg/mL. The mean recoveries are 95.4% to 98.1%. The results indicate that the HPLC method developed can easily be applied to the determination of eight polyphenols in the leaves of Crataegus pinnatifida Bge. var. major. PMID- 19298707 TI - Simultaneous determination of seven active flavonols in the flowers of Abelmoschus manihot by HPLC. AB - A high-performance liquid chromatography method is developed for the simultaneous quantification of seven flavonols, namely quercetin-3-O-robinobioside, hyperin, isoquercetin, hibifolin, myricetin, quercetin-3'-O-glucoside, and quercetin, in the flower of Abelmoschus manihot. These seven flavonols are selected as chemical markers because they are the major pharmacologically active constituents in the flower. The method involves the use of a Thermo ODS-2HYEPRSIL reversed-phase column (5 microm, 250 x 4.6 mm) at 25 degrees C with a mixture of acetonitrile and aqueous H(3)PO(4) as the mobile phase and detection at 370 nm. The recovery of the method is 94.31-107.08% with an RSD < or = 3.14% and the linearity (r(2) > 0.9996) is obtained for all the flavonoids. The current assay method can be readily utilized for the determination of the flavonols present in the flower and is considered to be suitable for the quality control of A. manihot samples. The comparison of flowers collected from nine locations shows that flavonoid glucoside is more stable than aglycon in the flower. This is the first study that analyzes the stability of flavonoids in the flower of A. manihot. This research also provides important evidence that the flower is a potentially abundant resource for obtaining hibifolin. PMID- 19298708 TI - Development of a reversed-phase high-performance liquid chromatographic method for analyzing furanocoumarin components in citrus fruit juices and Chinese herbal medicines. AB - A rapid and sensitive reversed-phase high performance liquid chromatographic method for the quantitation of five furanocoumarins (bergaptol, psoralen, bergapten, 6',7'-dihydroxybergamottin, and bergamottin) is developed and validated. HPLC analysis of these five furanocoumarins is performed on a reversed phase Inertsil ODS-2 column with a particle size of 5 microm. Using only water and acetonitrile as solvents, good separation, good precision, and high accuracy are obtained for the analysis of furanocoumarin components. This method is validated and applied to analyze the composition of five furanocoumarins in four citrus fruit juices (grapefruit, pomelo I, pomelo II, and shaddock) and ten Chinese herbal medicines (Bai-Zhi, Qiang-Huo, Du-Huo, Fang-Feng, Dang-Gui, Huang Qin, Gan-Cao, Chen-Pi, Ge-Gen, and Yin-Chen-Hao) prepared by water decoction or an alcohol infusion. Results show that four of the five furanocoumarins (but not bergapten) are detected in grapefruit, pomelo I, and pomelo II, and the highest amount of these components is found in grapefruit juice. In the ten Chinese herbal medicines, the five furanocoumarins are not detected in Ge-Gen or Yin-Chen Hao. The remaining herbs contain various compositions and amounts of furanocoumarins. In general, Chinese herbal medicines prepared by the 40% ethanol infusion contain larger amounts of furanocoumarins than those prepared by hot water decoction. PMID- 19298709 TI - Simultaneous quantitation of dicaffeoylquinic acids in rat plasma after an intravenous administration of mailuoning injection using liquid chromatography mass spectrometry. AB - A high-performance liquid chromatography electrospray ionization mass spectrometry method is developed and validated for the simultaneous quantitation of three major phenolic acids including 1,5-dicaffeoylquinic acid (1,5-DCQA), 3,4 dicaffeoylquinic acid (3,4-DCQA), and 3,5-dicaffeoylquinic acid (3,5-DCQA) in rat plasma. All analytes and internal standard (bergeninum) are extracted from plasma samples by liquid-liquid extraction with isopropanol. The chromatographic separation is accomplished on a stainless-steel column with a gradient 0.1% formic acid-acetonitrile solution as mobile phase at a flow rate of 0.2 mL/min with an operating temperature of 40 degrees C. The selected ion monitoring is performed at m/z 515.2 for 1,5-DCQA, 3,4-DCQA, and 3,5-DCQA, and m/z 327 for the internal standard bergeninum. Linear detection responses are obtained at a concentration range from 0.020 to 5.0 microg/mL for 1,5-DCQA, and 0.039 to 10.0 microg/mL for 3,4-DCQA and 3,5-DCQA. The lower limits of quantitation for 1,5 DCQA, 3,4-DCQA, and 3,5-DCQA are 20, 39, and 39 ng/mL, respectively. The intra- and inter-day precisions (RSD%) are within 11.0%, and the deviations of the assay accuracies are within +/- 12.0% for all analytes. The recoveries are greater than 84.0%. All analytes are proved to be stable during the sample preparation and analytic procedures. The method is successfully applied to the pharmacokinetic study of 1,5-DCQA, 3,4-DCQA, and 3,5-DCQA following an intravenous dose of 10 mL/kg mailuoning injection to rats. PMID- 19298710 TI - Development and validation of a reversed-phase HPLC method for separation and simultaneous determination of process-related substances of mirtazapine in bulk drugs and formulations. AB - A simple and rapid reversed-phase high-performance liquid chromatographic method has been developed for the separation and simultaneous determination of related substances of mirtazapine in bulk drugs and pharmaceutical formulations. Six impurities, including one degradation product of mirtazapine, have been separated on a BDS Hypersil (4.6 x 250 mm; particle size 5 microm) column with a mobile phase consisting of 0.3% triethylamine (pH 3.0)-acetonitrile (78:22 v/v) eluted in an isocratic mode and monitored with a photo diode array detector at 215 nm. The chromatographic behavior of all the analytes was studied under variable compositions of different solvent systems, temperatures, buffer concentrations, and pH values. The method was validated in terms of accuracy, precision, and linearity. The inter- and intra-day assay precision was found to be < 0.98% [relative standard deviation; (RSD)] and the recoveries were in the range 95.54 102.22% with RSD < 2.21%. The correlation coefficients for calibration curves for mirtazapine as well as impurities were in the range of 0.9941-0.9999. The method was successfully applied to the analysis of commercial formulations and the recoveries of mirtazapine were in the range of 99.38-100.73% with < 0.52% RSD. The method is useful not only for rapid evaluation of the purity of mirtazapine, but also for the simultaneous determination of related substances in bulk drugs and pharmaceutical formulations. PMID- 19298711 TI - Determination of tramadol, metamizole, ropivacaine, and bupivacaine in analgesic mixture samples by HPLC with DAD detection. AB - A high-performance liquid chromatography-diode array detection method was developed and validated to simultaneously determine tramadol (TMD), metamizole (MTZ), ropivacaine (RPV), and bupivacaine (BPV) in the presence of 4 methylaminoantypirine (4-MAA), the metabolite of MTZ, in analgesic mixtures samples used in Patient Controlled Analgesia (PCA). Chromatographic separation is achieved with a C-18 column using a mixture of ACN-methanol-water adjusted to pH 3.0 with NaH(2)PO(4) 0.05M (10:25:65 v/v) in an isocratic mobile phase at a flow rate of 0.8 mL/min. 0.5 mg/mL of Na(2)SO(3) in the water of the mobile phase was necessary to prevent the fast MTZ hydrolysis process to 4-MAA. Ultraviolet-diode array detection was used and chromatograms were registered at the wavelength of 230 nm. The method was linear in the range of 2.2-80.0 mg/L for TMD, 4.1-140.0 mg/L for MTZ, 2.3-40.0 mg/L for RPV, and 2.9-40.0 mg/L for BPV. Validation of the method was made in terms of accuracy, intra- and interday precision, as well as quantification and detection limits. The hydrolysis of MTZ to 4-MAA was studied and verified by mass spectrometry. The developed method was used successfully to evaluate the chemical stability of binary analgesic TMD mixtures with MTZ, RPV, or BPV. The mixtures were tested at standard concentrations used in PCA and in different storage conditions. When mixtures contained MTZ, a chromatographic peak from the metabolite 4-MAA was always detected in the chromatograms. PMID- 19298712 TI - Speciation of chromium metal ions by RP-HPLC. AB - A high-performance liquid chromatographic method with UV detection, based on chelation with morpholine-4-carbodithioate (MDTC), has been developed for the speciation of chromium. Simultaneous preconcentration of Cr(III) and Cr(VI) in aqueous solutions was achieved by extraction of metal species in chloroform as MDTC complexes. The separation of MDTC chelates of Cr(III) and Cr(VI) was realized on a C18 column with the use of acetonitrile-water (70:30) at a flow rate of 1.0 mL/min. The Cr(III) showed a single peak in chromatogram due to Cr(MDTC)(3), and Cr(VI) showed two peaks due to the formation of Cr(MDTC)(3) and Cr(MDTC)(2)(OMDTC). These two complexes were separated with different retention times at 5.4 and 4.3 min, respectively. The developed method can be used for rapid routine determination of chromium species with high precision and reliability. The method was validated by applying to various synthetic binary mixtures prepared by spiking Cr(III) and Cr(VI) in tap water. PMID- 19298713 TI - A harmless method for determining trenbolone acetate together with 17beta trenbolone in beef. AB - An inexpensive, harmless, and quick technique of sample preparation followed by a reversed-phase high-performance liquid chromatography (HPLC) for the simultaneous quantification of trenbolone acetate and 17beta-trenbolone in cattle muscle is presented. Analysis by HPLC with photo-diode array detector was performed using a short C1 column with an isocratic mobile phase. The method was validated by the analyses of spiked samples, resulting recoveries (> or = 87.9%; relative standard deviations < or = 3.4%), analytical total time (< 20 min/sample, where, a batch of 12 samples in 2 h), and quantitation limits (< or = 1.8 ng/g). No harmful organic solvents and reagents were used at all. PMID- 19298715 TI - Molecularly imprinted polymers: providing selectivity to sample preparation. PMID- 19298714 TI - Retention mechanism of selected ionic liquids on a pentafluorophenylpropyl polar phase: investigation using RP-HPLC. AB - This study investigated the retention mechanism of a congeneric group of imidazolium ionic liquid cations with alkyl functional groups of different lengths, an aryl substituent, and one pyridinium cation on a pentafluorophenylpropyl silica-based stationary phase. The influence of organic modifier type and content in the mobile phase on the retention mode of alkylimidazolium ionic liquid cations shows a 'U-shaped' relationship. The use of this stationary phase gives excellent linearity and detection limits in the ppt range. The method is also readily applicable to the highly selective analysis of plant extracts previously spiked with ionic liquids. PMID- 19298716 TI - Association between acute graft versus host disease and lung injury after allogeneic haematopoietic stem cell transplantation. AB - OBJECTIVE: To investigate the characteristics of chest high-resolution computed tomography (HRCT) and pathogenesis of acute graft versus host disease (acute GVHD)-induced lung injury after allogeneic haematopoietic stem cell transplantation (allo-HSCT). METHODS: A study of 47 patients with acute GVHD of grades II-IV describes the clinical manifestations and characteristics of chest HRCT of acute GVHD-induced lung injury. Detection of serum interferon gamma (IFNgamma) and tumour necrosis factor alpha (TNFalpha) were performed before the treatment for acute GVHD. Transbronchial biopsy was performed in four patients whose chest HRCT did not recover completely after treatment for acute GVHD. Pulmonary function was measured in patients who survived more than 6 months in every 3 months. RESULTS: Chest HRCT scans were performed in 47 cases and 20 cases showed abnormal in which 17 cases were suspected of acute GVHD-induced lung injury. In 17 patients with acute GVHD-induced lung injury, HRCT revealed diffused interstitial infiltrate in five cases, diffused interstitial and alveolar infiltrate in seven cases, diffused interstitial and segmental lobar alveolar infiltrate in five cases accompanied by bilateral pleural effusion and hydropericardium in nine patients. There was no statistical significance between the levels of serum IFNgamma and TNFalpha in cases with and without lung injury, but the levels of serum IFNgamma and TNFalpha in patients were significantly higher than the healthy group (IFNgamma: p=0.000, TNFalpha: p=0.000). The histopathology of the lung tissue was characterized by disorganization, epithelial cell damage, interstitial fibroplasias and interstitial T lymphocyte or macrophage infiltrate. Forty-seven cases all attained the treatment for acute GVHD, and the total effective rate and the rate of completely remission (CR) were 74.47 and 55.32%, respectively. The total effective rate and the rate of CR in the treatment for acute GVHD-induced lung injury were 94.12 and 58.82%, respectively. The effective rate of treatment for acute GVHD-induced lung injury positively correlated with that for acute GVHD (r=0.771, p=0.001). Three cases in nine cases with lung injury and three cases in 15 cases without lung injury who survived more than 6 months developed late-onset non-infectious lung injury. Eleven patients of 24 patients who survived more than 6 months had abnormal pulmonary function, including seven patients in nine patients with acute GVHD induced lung injury and four patients in 15 patients without acute GVHD-induced lung injury. There was no difference in the incidence of late-onset non infectious lung injury, but significance in the incidence of abnormal pulmonary function between cases with and without lung injury (p=0.033, cross-tabs). CONCLUSIONS: These results suggested that the lung might be one of the target organs of acute GVHD and participation of T lymphocyte, macrophage and cytokines such as IFNgamma and TNFalpha might play a role in the pathogenesis of acute GVHD induced lung injury. Acute GVHD-induced lung injury may progress to late-onset non-infectious lung injury. PMID- 19298717 TI - Reverse seroconversion of hepatitis B virus after allogeneic hematopoietic stem cell transplantation in the absence of chronic graft-versus-host disease. AB - The appearance of hepatitis B surface antigen (HBsAg) in patients previously positive for antibody to this antigen (HBsAb) is called reverse seroconversion, a rare complication after hematopoietic stem cell transplantation (HSCT), which occurs almost exclusively after HSCT from an HBsAb-negative donor and the development of chronic graft-versus-host disease (CGVHD). However, we experienced a patient who developed reverse seroconversion 23 months after unrelated HSCT even in the absence of immunosuppressants use or CGVHD. Serum immunoglobulin level was persistently normal. Therefore, all HBsAb-positive recipients should be considered to be at risk for HBV reactivation, even in patients without any risk factors. PMID- 19298718 TI - Does mobilization for autologous stem cell transplantation damage stromal layer formation? AB - Autologous hematopoietic stem cell transplantation (HSCT) has proved efficient to treat hematological malignancies. However, some patients fail to mobilize HSCs. It is known that the microenvironment may undergo damage after allogeneic HSCT. However little is known about how chemotherapy and growth factors contribute to this damage. We studied the stromal layer formation (SLF) and velocity before and after HSC mobilization, through long-term bone marrow culture from 22 patients and 10 healthy donors. Patients' SLF was similar at pre- (12/22) and post mobilization (9/20), however for controls this occurred more at pre-mobilization (9/10; p=0.03). SLF velocity was higher at pre than post-mobilization in both groups. Leukemias and multiple myeloma showed faster growth of SLF than lymphomas at post-mobilization, the latter being similar to controls. These findings could be explained by less uncommitted HSC in controls than patients at post mobilization. Control HSCs may migrate more in response to mobilization, resulting in a reduced population by those cells. PMID- 19298719 TI - Prognostic significance of immunohistochemical classification of diffuse large B cell lymphoma. AB - AIM: To evaluate the clinical significance of immunoperoxidase staining for CD10, bcl-6, mum-1 and bcl-2 to subdivide DLBCL into prognostic subgroups, we analysed 50 DLBCL cases using immunohistochemical methods. METHODS AND RESULTS: Fifty DLBCL patients were evaluated retrospectively. The expression of CD10 was associated with better OS (p=0.04) whereas expression of mum-1 was associated with worse OS (p=0.009). There were no significance of OS in case of expression of bcl-6 (p=0.05) and bcl-2 (p=0.3). They were subclassified using CD10, mum-1, bcl-6 as germinal center B-cell like (GCB) lymphoma (30%) and non-GCB lymphoma (70%). The OS and EFS (event free survival) were longer in GCB group (p=0.002) and 5-year OS for GCB group was 92% compared with only 44% for the non-GCB group (p=0.02). The OS of the GCB group also was longer compared to that of the non-GCB group in low IPI subgroup (p=0.01). CONCLUSION: The existence of survival differences between GCB a non-GCB group also in the patients with low IPI score, showed the importance of prognostic classification in the risk-adaptive treatment approaches. The classification as GCB and non-GCB based immunostains may enable to define more accurate prognostic groups in DLBCL. PMID- 19298720 TI - Fluorescent in situ hybridization studies in multiple myeloma. AB - Conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) results of bone marrow samples of 36 multiple myeloma (MM) patients at the time of diagnosis have been evaluated. Three probes for chromosome 13q (RB1, D13S319, D13S25), one for 14q32 (IgH) and one for 17p13 (p53) have been used for hybridization with fixed cells. Twenty patients (55.5%) had normal karyotypes, whereas eight (22.2%) had numerical or structural chromosomal abnormalities. We did not find metaphases for chromosome analysis in eight (22.2%) patients. Fluorescence in situ hybridization analyses revealed at least one or more abnormal results in 25 (69.5%) cases, whereas 11(30.5%) cases had no abnormal findings. 14q32 rearrangement was the most common finding in FISH analyses and has been detected in 21 cases (58.3%). 13q deletion and 17p deletion have been detected in 11 (30.5%) and 5 (13.9%) cases, respectively. Fluorescence in situ hybridization studies including 14q32 and 17p13 chromosome regions may yield quite significant results during clinical follow-up of MM. PMID- 19298721 TI - TCR zeta chain expression in T cells from patients with CML. AB - Chronic myeloid leukemia (CML) is a clonal disorder of the stem cells which has impaired cell-mediated immune response. Such immune dysfunction may be due to reduced expression of the T-cell receptor (TCR) zeta chain, which is an important component in TCR-mediated signal transduction. In this study, the TCR zeta expression level was detected using a real-time polymerase chain reaction with SYBR Green I technique and relative quantification method. We demonstrated that peripheral blood mononuclear cells (PBMCs) from patients with CML in chronic phase (CML-CP) and CML in complete remission (CML-CR) expressed decreased TCR zeta mRNA levels compared to healthy controls. In addition, TCR zeta chain gene expression was down-regulated in CD4+ T cells from patients with CML-CP. However, the mRNA expression level of the same gene in CD8+ T cells between CML patients and normal individuals did not differ. To confirm that this abnormal zeta chain expression could be corrected, we then stimulated the PBMCs of there patients with interleukin-2, phytohaemagglutinin and concanavalin A. The results showed that it was possible to partially up-regulate zeta chain expression. Taken together, these results indicate that TCR zeta chain expression was decreased in PBMCs and CD4+ T cells from patients with CML and this could be up-regulated partially by using stimulators. PMID- 19298722 TI - HLA-G expression in B chronic lymphocytic leukemia: a new prognostic marker? AB - Chronic lymphocytic leukemia (CLL) is characterized by a malignant clonal population of lymphocytes, which are usually of the B cell lineage. Classical Rai and Binet staging of CLL is being superseded by new prognostic markers. The mutational status of the immunoglobulin variable region heavy-chain genes segregates the disease into more benign and more malignant versions, and has been confirmed as an important prognostic marker in prospective clinical trials. A search for surrogate markers for this assay has led to flow cytometric assays for CD38 and ZAP-70 expression. The human leukocyte antigen G (HLA-G) molecule exhibits limited tissue distribution and a low polymorphism that generate seven HLA-G isoforms. HLA-G exerts multiple immunoregulatory functions. Recent studies indicate an ectopic up-regulation in tumor cells that may favor their escape from anti-tumor immune responses. For this report we studied HLA-G in parallel with CD38 and ZAP-70 in B-cell chronic lymphocytic leukemia (B-CLL) patients. HLA-G expression was studied retrospectively in circulating B-CLL cells from 20 patients by flow cytometry using the anti-HLA-G specific monoclonal antibody MEM/G9. The proportion of leukemic cells expressing HLA-G varied from 1 to 34%. We detected a statistically significant correlation between HLA-G positive (>12%) expression and progression free survival (p=0.045), but no correlation with CD38 and ZAP-70. We also detected a statistically significant difference between Binet stage A; B and C (p=0.046) and a positive correlation between IL-10 and HLA-G (p=0.044). We conclude that positive HLA-G has an effect on progression - free survival, when compared with CD38 and ZAP-70. PMID- 19298723 TI - The T-cell receptor Vbeta gene repertoire and clonal expansion from peripheral blood T cells in benzene-exposed workers in China. AB - The toxicity of benzene to lymphocytes is well known; however, little is known about the variety of T-cells that are rearranged after benzene exposure. In order to further understand the relative toxicity of benzene on T-cells, we investigated the features of the complementarity determining region 3 (CDR3) of T cells from benzene exposed workers. CDR3 of the T-cell receptor variable beta chain (TCRVbeta) subfamily genes were amplified using reverse transcriptase polymerase chain reaction. The polymerase chain reaction products were further subjected to genescan analysis to evaluate the clonality of T cells. Results revealed 2-12 Vbeta subfamilies identified in 16 benzene-exposed workers. The number of detectable Vbeta subfamilies present in the benzene-exposed group was significantly lower than in the control group (p<0.05). The most frequently used Vbeta genes were Vbeta3, Vbeta21, Vbeta6, and Vbeta16. We observed a skewed distribution and clonal expansion of TCR Vbeta subfamilies in benzene-exposed workers. PMID- 19298724 TI - Can mean platelet component be used as an index of platelet activity in stable coronary artery disease? AB - Acute coronary syndrome is associated with intracoronary thrombosis secondary to platelet activation. Previous groups have investigated platelet activation in both stable and unstable vascular disease. Most measures of platelet activation are not routinely available or easily adaptable to large scale clinical use. Recently, measurement of the mean platelet component (MPC) has become part of the routine data provided by an automated full blood count analyser, the Advia 120. MPC measures platelet density which changes on platelet activation. Our objectives were to determine if platelet activation, as measured by MPC, is increased in patients with stable coronary artery disease (CAD) and to determine if MPC could be useful in differentiating people with stable CAD from controls on an everyday clinical basis. Three hundred and forty-five consecutive patients attending for elective coronary angiography had full blood count analysis and MPC measurement performed using an ADVIA-120 analyser. Three hundred and twenty-four were analysed in our final dataset. Two hundred and fifty-three (78%) had CAD. Patients with CAD were significantly (p<0.001) older than those without (63.8 versus 56.0 years). Results failed to demonstrate a difference (p=0.467) in MPC between patients with CAD and those with normal coronary arteries (25.8 versus 26.0). Likewise, there was no correlation between MPC and the severity of CAD (Kendall's tau b=-0.086, p=0.04). MPC is not a useful index of platelet activity in stable CAD when used in everyday clinical practice. PMID- 19298725 TI - The effect of subclinical hypothyroidism on platelet parameters. AB - BACKGROUND: Hypothyroidism has a broad clinical spectrum. Today, physicians frequently encounter patients with very mild thyroid dysfunction instead of overt hypothyroidism. These patients have normal serum levels of thyroxine and triiodothyronine and only mildly elevated serum thyrotropin levels. Such patients are often identified through routine screening or in the course of an evaluation of common nonspecific symptoms. On the other hand, coronary heart disease is the leading cause of death in developed countries. There are studies, which suggest platelets play a role in the pathogenesis of atherosclerosis and coronary heart disease. AIM: The aim of this study is to compare the platelet count and other platelet parameters in subclinical hypothyroidic and euthyroidic healthy control group and to investigate whether these parameters have a predictive significance in patients with subclinical hypothyroidism. MATERIALS AND METHODS: Forty-seven patients with subclinical hypothyroidism and 30 euthyroidic healthy control group were enrolled into the study. RESULTS: Patients with subclinical hypothyroidism had higher mean platelet volume (MPV) and platelet distribution width (PDW) values than control group, which were statistically significant (p<0.001 and p<0.001), respectively. CONCLUSION: Our results indicate that MPV and PDW play an important predictive role in subclinical hypothyroidism. PMID- 19298726 TI - The effect of storage on the clinical utility of the immature platelet fraction. AB - The immature platelet fraction (%IPF) is a highly utilised new parameter in the full blood count (FBC) profile. It is a measure of the number of reticulated platelets and hence provides information on platelet transfusion requirements. It is common practice to measure %IPF values on blood samples that are several hours old, however unlike the well known good stability of other well established and more traditional FBC parameters such as haemoglobin and platelet counts, previous animal studies have shown that reticulated platelets are unstable. This study therefore assessed the validity of %IPF values obtained from stored patient samples. Our findings showed that %IPF values alter with storage; significant differences in %IPF values were obtained on stored samples compared with freshly analysed samples (p<0.001). It is therefore essential that in order to maintain the clinical utility of the %IPF, analyses should only be made on freshly drawn blood samples. PMID- 19298727 TI - Use of HbA estimation by CE-HPLC for prenatal diagnosis of beta-thalassemia; experience from a tertiary care centre in north India: a brief report. AB - Prenatal diagnosis (PND) of beta-thalassemia is offered by DNA based mutation analysis at 10-12 weeks gestation using chorionic villous biopsy specimens. Most centers offering PND search for five common mutations prevalent in India, which cover more than 90% of patients. However in those families where the mutation cannot be identified in the parents, PND becomes difficult by mutation detection. The technique thus followed is estimation of globin chain synthesis ratio in second trimester cord blood samples. However, this technique is cumbersome and not available in most of the centres. We evaluated 112 cord blood samples (74 were informative) for analyzing hemoglobin A (HbA) levels by cation-exchange high performance liquid chromatography (CE-HPLC) using the BioRad beta-thal short program between 18 and 22 weeks of gestation. A normal range for the population was calculated by running 14 cord blood samples of non-thalassemic abortuses of the same gestation. A 6 month post-natal follow-up was possible in 18 cases. All but one of these corroborated their PND. PMID- 19298728 TI - Microsurgical TESE versus conventional TESE for ICSI in non-obstructive azoospermia: a randomized controlled study. AB - In a population of non-obstructive azoospermia patients, the efficacy of microsurgical testicular sperm extraction (microTESE) and conventional TESE was evaluated in a randomized controlled study on 138 testicles, classified and paired in a 48-square table according to the different classes of the following three variables: patient plasma FSH concentration, orchidometry and testicular histology. Sperm retrieval was positive in 21/22 testicles with hypospermatogenesis (11/11, 10/11; microTESE, TESE respectively), in 12/14 with maturation arrest (6/7, 6/7), in 16/22 with incomplete Sertoli cell-only syndrome (8/11, 8/11), and in 16/80 with complete Sertoli cell-only syndrome (11/40, 5/40). Sperm recovery was positive in 5/24 patients with FSH concentration > or = 3 x maximum value of normal range (N) (4/12, 1/12), in 17/40 patients with 2N < or = FSH < 3N (9/20, 8/20), in 30/48 patients with N < FSH < 2N (17/24, 13/24), and in 13/26 patients with FSH = N (6/13, 7/13). Regarding orchidometry, sperm recovery was positive in 11/18 testicles with volume (V) > or = 12 ml (6/9, 5/9), in 27/56 testicles with 8 ml < or = V < 12 ml (15/28, 12/28), and in 27/64 testicles with V < 8 ml (15/32, 12/32). FSH value and the surgical procedure were the two variables significantly (P < 0.05) predicting positive sperm retrieval. PMID- 19298729 TI - Motile sperm organelle morphology examination is stricter than Tygerberg criteria. AB - The present study aimed to evaluate the correlation between the motile sperm organelle morphology examination (MSOME) and a well-known sperm morphology classification (Tygerberg criteria). For MSOME, spermatozoa were analysed at x8400 magnification by inverted microscope equipped with Nomarski differential interference contrast optics, Uplan Apo x100 oil/1.35 objective lens and variable zoom lens. By Tygerberg criteria, the semen underwent morphological evaluation as described in the literature. Regression analysis demonstrated significant positive correlation between percentage of normal sperm forms by Tygerberg criteria and by MSOME (r = 0.83, P < 0.0001). However, the incidence of normal spermatozoa by Tygerberg criteria (9.4%) was significantly higher (P < 0.0001) than under MSOME (3.3%). Despite the highly positive correlation, MSOME is a much stricter criterion of sperm morphology classification, since it identifies vacuoles and chromatin abnormalities that are not evaluated with the same precision by the analysis of Tygerberg criteria. MSOME should be included among the routine criteria for semen analysis. In addition, MSOME should be used for selection of spermatozoa for intracytoplasmic sperm injection based on the already published literature, as this is a good selection tool. PMID- 19298730 TI - Novel cause of hereditary obstructive azoospermia: a T2 allele in the CFTR gene. AB - Congenital bilateral absence of the vas deferens (CBAVD) is a frequent cause of obstructive azoospermia, and caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. A novel TG(13)T(2) allele was identified in a CBAVD patient with no clinical cystic fibrosis phenotype, normal pancreatic function, normal sweat chloride concentrations and no Y chromosome microdeletions. This case was studied for CFTR mutations, IVS8-poly(T), and M470V exon 10 missense polymorphism. One novel allele was detected in the (TG)(m)(T)(n) loci that had not been reported previously. This patient carried a [TG(11)T(9); R117H; p.Met470Val] haplotype on the other chromosome. Since the TG(13)T(2) allele was a compound heterozygote with R117H mutation, it was difficult to judge the severity of this allele. To better understand the complex regulation of exon 9 splicing, the levels of correctly spliced CFTR transcripts in CFTR-expressing epithelial cells derived from vas deferens and epididymis were analysed. These data emphasize the role of the T2 allele in CBAVD, and identify the T2 allele as a severe CBAVD disease-causing mutation. According to the data, the longer (TG)(m) polymorphic tract increases the proportion of transcripts with exon 9 deletion (9-), but only when activated by the short T allele. PMID- 19298731 TI - Ovarian stimulation in polycystic ovary syndrome patients: the role of body mass index. AB - In an attempt to examine whether body mass index (BMI) may influence IVF outcome in polycystic ovary syndrome (PCOS) patients undergoing ovarian stimulation with either gonadotrophin-releasing hormone (GnRH)-agonist (agonist group) or antagonist (antagonist group), 100 IVF cycles were studied: 35 in the agonist and 65 in the antagonist groups. In both agonist and antagonist groups, patients with BMI < or = 25 kg/m(2) had a significantly higher fertilization rate compared with patients with BMI > 25 kg/m(2) (P < 0.02 and P < 0.01, respectively). Lean patients (BMI < or = 25) undergoing ovarian stimulation using the GnRH-agonist, demonstrated the highest pregnancy rate. In conclusion, in this series of PCOS patients undergoing IVFembryo transfer cycles, ovarian stimulation utilizing the midluteal long GnRH-agonist suppressive protocol yielded a higher pregnancy rate in lean patients, probably due to its ability to lower the high basal LH milieu and its detrimental effect on oocyte quality and implantation potential. PMID- 19298732 TI - Receptors for thyroid-stimulating hormone and thyroid hormones in human ovarian tissue. AB - Dysfunction in thyroid regulation can cause menstrual and ovulatory disturbances, the mechanism of which is not clear. The distribution and activity of the thyroid stimulating hormone (TSHR), and the thyroid hormone receptors (TR) alpha1, alpha2 and beta1 in human ovarian tissue and in granulosa cells was studied using immunohistochemistry, reverse-transcriptase polymerase chain reaction (RT-PCR), quantitative PCR and immunoassays. Strong immunostaining of TSHR, TRalpha1 and TRbeta1 was observed in ovarian surface epithelium and in oocytes of primordial, primary and secondary follicles, with minimal staining in granulosa cells of secondary follicles. Granulosa cells of antral follicles expressed TSHR, TRalpha1 and TRbeta1 proteins. Messenger RNA for all receptors was present in ovarian tissue. Mature human granulosa cells expressed transcripts for 5' deiodinases types 2 and 3, but not type 1, indicating the possibility of conversion of peripheral thyroid hormone thyroxin (T(4)). Granulosa cells stimulated with TSH showed a significant increase in cAMP concentrations after 2 h of culture (P = 0.047), indicating activation through TSHR. Stimulation with T(4) resulted in increased extracellular signal-regulated kinase 1 and 2 activation after 10, 30, 60 min and 24 h. These data demonstrate that TSH and thyroid hormone receptors may participate in the regulation of ovarian function. PMID- 19298733 TI - Individual ovarian volumes obtained from 2-dimensional and 3-dimensional ultrasound lack precision. AB - Measuring ovarian volume has been suggested as a possible screening test to assess a woman's ovarian reserve. For such a screening tool to be clinically useful, knowledge of its precision and reproducibility is essential. Recent advances in ultrasound scanning techniques allow the measurement of volumes in three dimensions rather than the traditional estimation from two dimensions. Transvaginal 2-dimensional (2D) and 3-dimensional (3D) ultrasound examinations were performed on 49 women attending a tertiary centre for investigation or treatment for subfertility between January and May 2006. Two observers calculated ovarian volume using both 2D (prolate ellipsoid formula) and 3D techniques [virtual organ computer-aided analysis (VOCAL)] with rotation steps of 30 degrees (3D-30). For the four comparisons (inter- and intra-observer; 2D and 3D-30) intraclass coefficients of 0.97 to 0.98, and standard errors ranging from 17% to 14% (for inter-observer 2D and intra-observer 3D, respectively) were obtained. The corresponding coefficients of repeatability ranged from 33% to 28%. These results suggest that measurement of transvaginal ovarian volumes using both 2D and 3D ultrasound is imprecise for individuals. The imprecision is greater for lower ovarian volumes, which may be important in clinical practice. The average of two or more measurements is likely to be more accurate than a single measurement. PMID- 19298734 TI - Efficiency of human oocyte slow freezing: results from five assisted reproduction centres. AB - It has been demonstrated previously that freezing oocytes within 2 h of retrieval increases the efficiency of cryopreservation via a slow-freezing/rapid-thawing protocol with 0.3 mol/l sucrose (SF/RT 0.3). The aim of this multicentre survey was to verify this observation on a larger scale. This was a retrospective study on the clinical outcome of 510 SF/RT 0.3 cycles divided into two groups: group A, freezing oocytes within 2 h of retrieval; group B, freezing oocytes more than 2 h after retrieval. The rate of best-quality embryos was significantly higher (33.24%) in group A than in group B (16.20%, P < 0.001). Pregnancy and implantation rates were 30.07% and 15.08% in group A versus 8.97% and 4.57% in group B (P < 0.001). Clinical pregnancy rates per thawed and per injected oocyte in group A were 5.53% and 10.41%, versus 1.46% and 2.77% in group B (P < 0.001). The overall yield from oocytes cryopreserved within 2 h of retrieval (group A) was 6.49 implantations per 100 oocytes thawed versus 1.74 for group B (P < 0.001). Embryo quality, pregnancy and implantation rates, and clinical efficiency of thawing cycles were all significantly improved when cryopreservation was carried out within 2 h of oocyte retrieval. PMID- 19298735 TI - Elevated NaCl concentration improves cryotolerance and developmental competence of porcine oocytes. AB - High hydrostatic pressure has been reported to improve the fertilizing or developmental ability of mammalian spermatozoa, oocytes and embryos. This study investigated the effect of another stress, temporarily increased NaCl concentration, on cryotolerance and developmental competence of porcine oocytes. In Experiment 1, survival rates were compared after 1 h exposure to seven elevated NaCl concentrations and 1 h recovery time. In Experiment 2, oocytes were exposed to 593 and 1306 mOsmol NaCl, subsequently recovered, vitrified, then subjected to parthenogenetic activation. Both cleavage and blastocyst rates increased after NaCl treatment compared with untreated controls. In Experiment 3, oocytes were treated with 593 mOsmol NaCl followed by 1 and 2 h recovery, respectively, then used as recipients for somatic cell nuclear transfer (SCNT). Cleavage rates were not different from those in untreated controls, but blastocyst rates increased in both NaCl-treated groups. In conclusion, treatment of porcine oocytes with elevated NaCl concentrations improved their developmental competence after vitrification and parthenogenetic activation or SCNT. Further experiments are required to investigate in-vivo consequences, and the effect on gametes and embryos of different mammalian species. PMID- 19298736 TI - Pronuclear morphology differs between women more than 38 and women less than 30 years of age. AB - Pronuclear morphology has been reported as a good tool for studying embryo development and euploidy. Comparing two groups of women with different aneuploidy risk, women more than 38 years old (n = 28) known to be at high risk of aneuploidy, and women under 30 years old (n = 35), this study investigated whether pronuclear morphology could be used routinely as an alternative to preimplantation genetic screening (PGS) in countries where PGS is prohibited. Pronuclear morphology was evaluated for 301 zygotes and related to embryo quality and pregnancy outcome. For the older women, an increased frequency of zygotes with abnormal polar body and pronuclei alignment was observed, i.e. type gamma, with 93% aneuploidy risk (26.0 versus 15.1% P < 0.05) and fewer zygotes with a good development prognosis (36.4 versus 47.8%; P < 0.05). A1alpha configuration was associated with good implantation rate and was not related to day 2 embryo quality. This configuration was less frequent in the group of women more than 38 years old and among non-pregnant women under 30 years, compared with pregnant women under 30 years old. Pronuclear morphology seemed linked to age, but not associated with embryo quality. A larger study allowing correlation analysis is necessary to confirm the value of these criteria and the link to a woman's age. PMID- 19298737 TI - Soluble HLA-G in IVF/ICSI embryo culture supernatants does not always predict implantation success: a multicentre study. AB - Several reports have described an association between the presence of soluble human leukocyte antigen G (sHLA-G) in human embryo culture supernatants (ES) and implantation success. However, not all studies agree with these findings. To further document this debate, a multicentre blinded study was performed to investigate, on a large number of IVF ES and ICSI ES, whether sHLA-G is a useful criterion for embryo selection before transfer. A total of 1405 ES from 355 patients were collected from three assisted reproductive technique (ART) centres and evaluated for their sHLA-G content in a single laboratory, using a chemiluminescence enzyme-linked immunosorbent assay. In only one centre was a significant association between sHLA-G-positive ES and successful implantation established (P = 0.0379), whereas no such association was observed in the other centres. It was found that the percentages and concentrations of sHLA-G-positive ES varied between centres, depending on culture media and ART conditions. The percentage of sHLA-G-positive ES was significantly higher in IVF ES than ICSI ES (P < 0.001 and P < 0.01 for two centres). These data demonstrate that substantial variations of sHLA-G content in ES occur between different ART centres, highlighting the influence of several technical parameters that differ from one centre to another. PMID- 19298738 TI - Assisted fertilization and embryonic axis formation in higher primates. AB - In naturally fertilized embryos of various organisms, the spermatozoon provides a localized cue to initiate early embryonic patterning. In mice, the sperm entry point (SEP) may reorient the first cleavage division, which separates the zygote into two halves that follow distinct fates. However, it is unknown whether the mechanical injection of spermatozoa into an oocyte by intracytoplasmic sperm injection (ICSI), a technique commonly used in human assisted reproduction, possesses such a role. Rhesus macaque embryos fertilized by ICSI were examined in order to determine the consequences of placing the spermatozoon at specific positions in the ooplasm and whether this can provide new information about patterning in mammalian eggs. The SEP specified by the injected spermatozoa was most often localized near the first cleavage plane and was mainly distributed along the boundary zone that separates the embryonic and abembryonic parts of the monkey blastocyst. Moreover, the ICSI data, when compared with naturally fertilized mouse embryos, showed a similar outcome in terms of cleavage axes and first embryonic axis specification. As there are no studies to date regarding sperm entry in human oocytes and its influence on embryonic development, this investigation using the rhesus macaque as a clinical model is noteworthy. PMID- 19298739 TI - Improving clinical understanding of the effect of ovarian stimulation on women's lives. AB - A conceptual model was developed of the impact of ovarian stimulation on women's functioning and wellbeing. A patient-reported outcome (PRO) measure was generated based on this model. Qualitative measures used comprised a discussion guide and coding system devised according to the Food and Drug Administration guidelines for developing PRO measures. Data were gathered through telephone and face-to face interviews and focus groups. A total of 47 women across nine fertility healthcare providers in France, UK and USA were included in the analysis. The mean age of patients was 35 years (range 24-44). Based on data collected and prior postulated domains, a conceptual model of the impact of ovarian stimulation was developed. Three major (psychological, interference in daily life, logistics) and one minor (side effects) domain were identified. Short-term consequences included problems with psychological health and productivity. Longer term consequences included depression and poor self-image. Factors that modified the impact of ovarian stimulation were also identified. This is the first model providing insight into the impact of ovarian stimulation on women's functioning and wellbeing. Once validated, it can be applied in quantitative research to improve understanding of the impact of ovarian stimulation, and assist in developing more patient-centred approaches to treatment. PMID- 19298740 TI - Is the dimension of ovarian endometriomas significantly modified by IVF-ICSI cycles? AB - Information regarding the growth and development of endometriomas during IVF intracytoplasmic sperm injection (ICSI) cycles is lacking. In this study, the aim was to estimate the influence of IVF-ICSI on the dimension of these cysts and to assess whether this treatment contributes to the manifestation of new endometriomas in patients already affected. Women were eligible if they had been diagnosed with ovarian endometrioma(s). Recruited patients who failed to become pregnant were scanned again 3-6 months later to evaluate the modification of the dimension of the cysts and/or the manifestation of new endometriomas. Forty-eight women completed the study protocol. The median (interquartile range) values for the volume of the cysts before and after the IVF-ICSI cycle were 3.9 (2.9-7.9) ml and 4.9 (2.4-9.9) ml, respectively (Wilcoxon rank test for paired data, not significant). The development of a new endometrioma was documented in one case (2.1%, 95% confidence interval 0.1-11.1%). Fear about the growth of ovarian endometriomas in women who have to undergo IVF-ICSI is not justified. PMID- 19298741 TI - Circulating cell-free DNA as a potential biomarker for minimal and mild endometriosis. AB - It has recently been reported that high concentrations of circulating cell-free (ccf) nucleic acids in plasma and serum could be used as biomarkers for non invasive monitoring a wide variety of malignant and benign proliferations and inflammatory conditions. Endometriosis is one of the most common benign gynaecological proliferations with inflammatory activation in premenopausal women. Real-time multiplex polymerase chain reaction was used for synchronized quantification of the glyceraldehyde-3-phosphate dehydrogenase gene sequence in nuclear DNA (nDNA) and the ATP synthase-8 gene sequence in mitochondrial DNA (mtDNA). DNA was extracted from 500 microl serum and plasma of 19 cases with endometriosis to measure the total amount of ccf nDNA and ccf mtDNA. The concentration of ccf nDNA in plasma was significantly higher in the endometriosis group than in the control group (P = 0.046). The cut-off value selected by a receiver operating characteristic curve could provide a sensitivity of 70% and a specificity of 87% to discriminate between the minimal or mild cases and normal controls. The finding of significantly increased concentrations of ccf nDNA in plasma of patients with endometriosis suggests that ccf nDNA might be a potential biomarker for developing non-invasive diagnostic test in endometriosis. PMID- 19298742 TI - Hysteroscopic polypectomy without cycle cancellation in IVF cycles. AB - Endometrial polyps destroy the endometrial texture and play an important role in implantation failure. There is no consensus about the management of patients diagnosed with endometrial polyp in IVF cycles. In this study, nine patients who underwent assisted reproduction treatment cycles were diagnosed with endometrial polyps less than 1.5 cm by transvaginal ultrasonography. Eight patients were treated by long protocol and one patient was the recipient of an egg donation cycle. In all patients, polyp resection was performed through hysteroscopic polypectomy. Polypectomy was done during ovarian stimulation in the standard treatment cycles, and during hormone replacement therapy in the recipient of the egg donation cycle. The interval between polyp resection and embryo transfer was 2-16 days. Four patients achieved pregnancy (two twins, two singletons), four patients were unsuccessful, and one pregnancy was a blighted ovum. All of the successful pregnancies are still ongoing. There is a dilemma regarding the management of patients diagnosed with endometrial polyps in assisted reproduction treatment cycles. If polypectomy before embryo transfer in an IVF cycle is proven to be safe, then embryos will be transferred without cycle cancellation. This study included nine patients; further studies with more patients are required to confirm these findings. PMID- 19298743 TI - ICSI outcome following conservative fertility sparing management of endometrial cancer. AB - Approximately 5% of cases of endometrial cancer occur in women aged 40 years or younger. In the presence of early staged endometrial cancer, fertility sparing management may be considered in selected patients. Following high dose progestin therapy and confirmation of the regression of cancer, the patient might attempt to conceive spontaneously. However, assisted reproduction techniques might increase the likelihood of pregnancy and decrease the time interval to conception. In this report, the authors present four patients with endometrial cancer who were treated conservatively with high dose progestin. In all of the cases, the endo-myometrial junctional zone was intact with no evidence of extra uterine spread on pelvic imaging. A total of seven intracytoplasmic sperm injection and embryo transfer cycles were performed in four patients; five healthy infants were delivered. Two additional spontaneous pregnancies occurred in two patients. During the follow-up period, no recurrence was noted. Although there are limited data, fertility sparing with high dose progestin therapy may be offered to patients with early stage disease and subsequently assisted reproductive techniques may be employed to achieve immediate pregnancy. PMID- 19298744 TI - Pelvic splenosis--a gynaecological challenge. AB - Pelvic splenosis is a post-traumatic spleen autotransplantation, which can mimic various gynaecological diseases. A case of a 39-year-old woman, who underwent partial splenectomy because of a road accident in her childhood, currently presenting with abdominal pain and suspected corpus luteus torsion. Diagnostic laparoscopy was conducted which ruled out twisted ovary. This confirmed disseminated pelvic splenosis as a coincidental finding. Following this procedure, the abdominal pain ceased. The current report aims to raise the awareness of splenosis so that it may be appropriately recognized and differentiated from other pelvic findings. PMID- 19298745 TI - Cytogenetic studies in couples with recurrent miscarriage in the Sultanate of Oman. AB - Miscarriage, defined as spontaneous pregnancy loss at <20-28 weeks' gestation, is a common clinical problem. Balanced chromosomal rearrangements in either parent are an important cause of repeated pregnancy loss, particularly in the first trimester. In this study, chromosomal abnormalities that cause recurrent miscarriage were evaluated in Omani parents and some of their dysmorphic children. A total of 380 couples (760 individuals) with two or more recurrent miscarriages were examined for chromosomal aberrations during the period 1999 2006. For each proband the chromosomal preparations were analysed and karyotyped after applying a Giemsa-trypsin banding method. The overall incidence of chromosomal anomaly was 26 out of 760 individuals (3.42%). These abnormalities included 21 (2.8%) structural aberrations and 5 (0.7%) numerical anomalies. In addition to these abnormalities, 39 (5.1%) chromosomal variants were also found. The nature of these abnormalities and their relation to obstetric history are discussed. In conclusion, chromosomal abnormality is one of the causes of recurrent miscarriage. This study illustrates the incidence and distribution of chromosomal abnormalities among Omani couples with recurrent miscarriage. Cytogenetic findings could provide valuable information for genetic counselling and allow monitoring of future pregnancies by prenatal diagnosis in couples with a history of recurrent miscarriage. PMID- 19298746 TI - Study of p53 gene mutations and placental expression in recurrent miscarriage cases. AB - This study aimed to investigate the role of p53 in early human development by screening patients with recurrent miscarriages (RM) for mutations in the p53 gene and by studying p53 expression in placental tissue. A total of 46 women with RM and 191 control women were included in the study. A sample was also obtained from 40 male partners of RM patients. The samples were screened for p53 sequence variations using denaturing high-performance liquid chromatography, sequencing and allele-specific polymerase chain reaction. Placental tissue was available from 19 miscarriages. p53 expression in placental tissue was studied by immunohistochemical staining. The C11992A polymorphism in p53 was found to be associated with RM in Finnish patients. The C/A or A/A genotype was detected in 32.6% of the women with RM and in 18.9% of the controls (P = 0.0414, odds ratio 2.083, confidence interval 1.018-4.259). The results suggest that women carrying the C/A or A/A genotype have a two-fold higher risk for RM than women with the C/C genotype. Further studies are, however, necessary to define whether the intronic polymorphism has functional consequences. The immunohistochemical staining of placental tissues revealed no abnormal p53 expression patterns in the samples studied. PMID- 19298747 TI - Metformin and atorvastatin reduce adhesion formation in a rat uterine horn model. AB - The aim of the present study was to determine whether atorvastatin and metformin are effective in preventing adhesions in a rat uterine horn model. A total of 40 non-pregnant, female Wistar albino rats, weighing 180-210 g, were used as a model for post-operative adhesion formation. The rats were randomized into four groups after seven standard lesions were inflicted in each uterine horn and lower abdominal sidewall using bipolar cauterization. The rats were given atorvastatin 2.5 mg/kg/day, p.o. (10 rats), atorvastatin 30 mg/kg/day, p.o. (10 rats), metformin 50 mg/kg/day, p.o. (10 rats) and no treatment was applied in the control group (10 rats). The animals were killed 2 weeks later and adhesions were scored both clinically and pathologically by authors blinded to groups. One rat in the control group died before the end of the 2 week period. Total clinical adhesion scores regarding extent, severity and degree of adhesions and histopathological findings including inflammation and fibrosis were significantly lower in the metformin (P < 0.001 and P < 0.01, respectively) and atorvastatin 30 mg/kg/day (P < 0.001 and P < 0.01, respectively) groups when compared with control group. Metformin and atorvastatin are both effective for prevention of adhesion formation in a rat uterine horn model. PMID- 19298748 TI - Oocyte-like structures arising from cells of follicular fluid are not captured in aspirates. AB - Recently, cells from ovarian surface epithelium (OSE) of post-menopausal women and women with premature ovarian failure were investigated and oocyte-like cells with diameters up to 95 microm were found to arise after a certain time in culture. In addition, it seems that a mixed population of germ cells and germline stem cells exists in non-follicle ovarian structures. Relating to an earlier publication, where it was shown that pre-antral follicles with immature oocytes could be captured in follicular fluid (FF) aspirates due to the incorporated tissue in the puncture needle, it was reasoned that OSE or otherwise germline stem cells, possibly captured equally through ovarian puncture, might give rise to oocyte-like cells. The aim of this study was therefore to try to derive such oocyte-like cells from FF aspirates of patients undergoing IVF after culture. Additionally, FF-derived cells were aggregated with human embryonic stem cells to see if an embryonic environment had the ability to enable cells from the FF aspirate to acquire an oocyte-like morphology. Investigations could not confirm the development of oocyte-like cells from cells of FF aspirates. PMID- 19298749 TI - Molecular neurosciences. PMID- 19298750 TI - Hyperbaric oxygen therapy and cerebral ischemia: neuroprotective mechanisms. AB - INTRODUCTION: Numerous studies have demonstrated a protective effect of hyperbaric oxygen therapy in experimental ischemic brain injury, and many physiological and molecular mechanisms of hyperbaric oxygen therapy-related neuroprotection have been identified. METHODS: Review of articles pertaining to hyperbaric oxygen therapy and cerebral ischemia in the National Library of Medicine and National Institutes of Health database, emphasizing mechanisms of hyperbaric oxygen therapy-related neuroprotection. RESULTS: Hyperbaric oxygen therapy has been shown to ameliorate brain injury in a variety of animal models including focal cerebral ischemia, global cerebral ischemia, neonatal hypoxia ischemia and subarachnoid hemorrhage. Small human trials of hyperbaric oxygen therapy in focal ischemia have not shown benefit, although one trial of hyperbaric oxygen therapy before cardiopulmonary bypass demonstrated improved neuropsychological and inflammatory outcomes with hyperbaric oxygen therapy. Hyperbaric oxygen therapy is associated with improved cerebral oxygenation, reduced blood-brain barrier breakdown, decreased inflammation, reduced cerebral edema, decreased intracranial pressure, reduced oxidative burden, reduced metabolic derangement, decreased apoptotic cell death and increased neural regeneration. CONCLUSION: On a molecular level, hyperbaric oxygen therapy leads to activation of ion channels, inhibition of hypoxia inducible factor-1alpha, up regulation of Bcl-2, inhibition of MMP-9, decreased cyclooxygenase-2 activity, decreased myeloperoxidase activity, up-regulation of superoxide dismutase and inhibition of Nogo-A (an endogenous growth-inhibitory factor). Ongoing research will continue to describe the mechanisms of hyperbaric oxygen therapy-related neuroprotection, and possibly expand hyperbaric oxygen therapy use clinically. PMID- 19298751 TI - Neuroprotective roles and mechanisms of neuroglobin. AB - OBJECTIVE: The objectives of this work were to update and summarize recent experimental works on neuroglobin, mainly focus on its neuroprotective effects and the mechanisms. METHODS: The literature was reviewed using PubMed database, and some of the recent findings from our laboratory were included. RESULTS: Neuroglobin is a recently discovered tissue globin with a high affinity for oxygen and is widely and specifically expressed in neurons of vertebrate's central and peripheral nervous systems. Investigations in the past several years have advanced our knowledge on the functions and mechanisms of neuroglobin, but many issues remain unclear. Emerging reports have shown that overexpression of neuroglobin confers neuroprotection against neuronal hypoxia or ischemia-induced damage in cultured neurons and in cerebral ischemic animal models. Accumulating findings suggest several possible neuroprotective roles and mechanisms including ligand binding and oxygen sensing, modulation of cell signaling pathways and maintenance of mitochondria function. CONCLUSION: Emerging experimental works suggest that neuroglobin is neuroprotective against hypoxic/ischemic insults, probably via ligand binding and oxygen sensing, modulation of cell signaling pathways and maintenance of mitochondria function. PMID- 19298752 TI - Neuroprotective effect of volatile anesthetic agents: molecular mechanisms. AB - INTRODUCTION: Intra-operative cerebral ischemia can be catastrophic, and volatile anesthetic agents have been recognized for their potential neuroprotective properties since the 1960s. In this review, we examine the neuroprotective effects of five volatile anesthetic agents in current or recent clinical use: isoflurane, sevoflurane, desflurane, halothane and enflurane. METHODS: A review of publications in the National Library of Medicine and National Institutes of Health database from 1970 to 2007 was conducted. RESULTS: Volatile anesthetic agents have been shown to be neuroprotective in multiple animal works of ischemic brain injury. Short-term neuroprotection (<1 week post-ischemia) in experimental cerebral ischemia has been reported in multiple works, although long-term neuroprotection (> or = 1 week post-ischemia) remains controversial. Comparison works have not demonstrated superiority of one specific volatile agent over another in experimental models of brain injury. Relatively few human works have examined the protective effects of volatile anesthetic agents and conclusive evidence of a neuroprotective effect has yet to emerge from human works. CONCLUSION: Proposed mechanisms related to the neuroprotective effect of volatile anesthetic agents include activation of ATP-dependent potassium channels, up regulation of nitric oxide synthase, reduction of excitotoxic stressors and cerebral metabolic rate, augmentation of peri-ischemic cerebral blood flow and up regulation of antiapoptotic factors including MAP kinases. PMID- 19298753 TI - Genes associated with idiopathic epilepsies: a current overview. AB - OBJECTIVE: This article aimed to review the latest genes associated with idiopathic focal and generalized epilepsies. METHODS: PubMed and Entrez Gene searches pertaining to this work was conducted using specific keyword search terms related to genes and various listed subtopics related to idiopathic epilepsy syndromes. RESULTS: Mutations in the cholinergic receptor, neuronal nicotinic, alpha2, alpha4 and beta2 subunit genes have been found in autosomal dominant nocturnal frontal lobe epilepsy. Mutations of potassium voltage-gated channel, KQT-like subfamily, members 2 and 3 genes were identified to be responsible for benign familial neonatal seizures. The voltage-gated sodium channel genes and gamma-aminobutyric acid receptor alpha subunit genes may be involved in the pathogenesis of generalized epilepsy with febrile seizure plus. Mutations of gamma-aminobutyric acid receptor alpha1, gamma-aminobutyric acid receptor delta, calcium channel voltage-dependent beta4 subunit and chloride channel 2 gene are associated with juvenile myoclonic epilepsy. In addition, mutations of leucine-rich, glioma-inactivated 1 gene leads to genetic abnormalities of familial lateral temporal lobe epilepsy. EF-hand domain (C terminal)-containing 1 gene can cause some patterns of juvenile myoclonic and juvenile absence epilepsies. DISCUSSION: Genetic factors play an important role in idiopathic epilepsy syndromes. Ion channel genes and some non-ion channel genes contribute to the pathogenesis of idiopathic epilepsies. Based on these findings, genetic diagnosis and new treatment strategies to part of idiopathic epilepsies become possible in the future. PMID- 19298754 TI - Characteristics and functions of NG2 cells in normal brain and neuropathology. AB - OBJECTIVE: This review is focused on the current understanding of the roles of the fifth class of non-neuronal cells, NG2 cells, in the central nervous system (CNS). METHODS: We have reviewed some literature on properties of NG2 cells, including cell morphology, expression of receptors and possible functions. RESULTS: Chondroitin sulfate proteoglycan (NG2) is expressed in a high proportion in non-neuronal cells of the CNS. During development, NG2 cells can differentiate into oligodendrocytes, astrocytes and neurons. In the adult, the NG2 cells have a common morphology: multibranched processes and small cell bodies, and are ubiquitously distributed throughout brain parenchyma. They possess some functional receptors and contact neurons at nodes of Ranvier or via synaptic terminals. Some NG2 cells can even fire action potential. Various brain injury models have demonstrated that NG2 cells adjacent to the damage site could increase in number and become hypertrophic. However, there is no clear evidence indicating the function of NG2 cells in the adult brain. DISCUSSION: The function of NG2 cells in the adult brain is still uncertain. The NG2 expressing cells may be progenitor cells in the developing brain. In the adult, the discovery of functional receptors, interactions with neurons and ability to respond to different harmful stimulations have implied roles of NG2 cells in facilitating neuronal network function, which may be important in brain inflammation, neurodegeneration and neuroregeneration. PMID- 19298755 TI - Cerebral vasospasm following subarachnoid hemorrhage: time for a new world of thought. AB - OBJECTIVE: Delayed cerebral vasospasm has long been recognized as an important cause of poor outcome after an otherwise successful treatment of a ruptured intracranial aneurysm, but it remains a pathophysiological enigma despite intensive research for more than half a century. METHOD: Summarized in this review are highlights of research from North America, Europe and Asia reflecting recent advances in the understanding of delayed ischemic deficit. RESULT: It will focus on current accepted mechanisms and on new frontiers in vasospasm research. CONCLUSION: A key issue is the recognition of events other than arterial narrowing such as early brain injury and cortical spreading depression and of their contribution to overall mortality and morbidity. PMID- 19298756 TI - Quetiapine regulates neurogenesis in ischemic mice by inhibiting NF-kappaB p65/p50 expression. AB - OBJECTIVES: Previously, we showed that quetiapine, an atypical antipsychotic drug, significantly attenuated neurodegeneration induced by global cerebral ischemia (GCI). The present work investigates the effects of quetiapine on neurogenesis. METHODS: Mice were treated with quetiapine (10 or 20 mg/kg/day; intraperitoneal injection) for 2 weeks and then subjected to GCI on day 15. Seven days after GCI, the mice were killed. Neuronal injury and neurogenesis were analysed using hematoxylin-eosin and 5-bromo-20-deoxyuridine stainings. Levels of nuclear factor kappaB (NF-kappaB) p65/p50 expressions were determined by immunohistochemistry and Western blot analysis. RESULTS: Global cerebral ischemia resulted in neuronal injury, neurogenesis and NF-kappaB p65/p50 expressions in hippocampus, especially in the dentate gyrus. Pre-administration of quetiapine significantly alleviated neuronal injury, while inhibiting neurogenesis and down regulating NF-kappaB p65/p50 expression. DISCUSSION: NF-kappaB plays a key role in regulating neuron damage and neurogenesis. This work suggests that down regulation of NF-kappaB expression may be one of the mechanisms by which quetiapine inhibits neurogenesis. PMID- 19298757 TI - Granulocyte colony-stimulating factor protects the brain against experimental stroke via inhibition of apoptosis and inflammation. AB - OBJECTIVES: The molecular mechanisms of the anti-apoptotic and anti-inflammatory properties of granulocyte-colony stimulating factor (G-CSF) following focal cerebral ischemia in rats were examined in this study. METHODS: Sprague-Dawley rats were randomly divided into three groups: sham, middle cerebral artery occlusion (MCAO) non-treatment and MCAO with G-CSF treatment. Focal ischemia was induced with the suture occlusion method for 90 minutes, and treatment was given at the onset of reperfusion. All animals were killed 24 hours after reperfusion. Assessment included neurological scores, infarction volumes, histology, immunofluorescent staining and Western blotting. RESULTS: G-CSF significantly reduced the infarct volume and ameliorated the early neurological outcome scores. Western blot analysis showed that G-CSF treatment significantly elevated the cIAP2 levels and decreased the activation of caspase 3 in the ischemic cortex compared with the non-treated rats. Immunofluorescent works also showed that G CSF treatment inhibited both neuronal and glial tumor necrosis factor alpha and interleukin 1beta expressions. DISCUSSION: The neuronal anti-apoptotic action of G-CSF may be mediated in part by the anti-apoptotic protein cIAP2. G-CSF also exerts anti-inflammatory actions after focal cerebral ischemia by preventing both neuronal and glial pro-inflammatory cytokine expressions. PMID- 19298758 TI - Effect of gap junction inhibition on intracerebral hemorrhage-induced brain injury in mice. AB - It has been reported that gap junction contributes to ischemic brain injury and gap junction inhibitors improve neurological outcome in ischemic brain injury models. In the present study, we investigated the effects of gap junction inhibitor, carbenoxolone, on mortality, neurological deficits and brain edema in mice with intracerebral hemorrhage. A total of 80 male CD-1 mice were divided into two parts with two end-points for this study. In part one, animals were divided into four groups: sham, vehicle treatment following intracerebral hemorrhage induction, low-dose carbenoxolone (33 mg/kg) treatment 1 hour after intracerebral hemorrhage induction and high-dose carbenoxolone (100 mg/kg) treatment 1 hour after intracerebral hemorrhage induction groups. Animals were euthanized after 24 hours. In part two, animals were divided into four groups: sham, vehicle treatment 1 hour after intracerebral hemorrhage induction, single high-dose of carbenoxolone treatment at 1 hour after intracerebral hemorrhage induction and three high-doses of carbenoxolone treatment 1, 24 and 48 hours respectively after intracerebral hemorrhage induction. Animals were euthanized after 72 hours. Intracerebral hemorrhage was induced by collagenase injection. Neurological deficits were evaluated using modified Garcia's neurological test, wire hanging and beam balance tests. Brain edema was measured by brain water content. Our results showed that intracerebral hemorrhage produced brain edema and neurological deficits in mice. Carbenoxolone treatment failed to reduce brain edema and neurological deficits. In fact, the high dose of carbenoxolone aggravated neurological deficits and increased mortality 72 hours after the treatment. In conclusion, inhibition of gap junction has no short-term neuroprotective effect on intracerebral hemorrhage-induced brain injury. Further studies are required to assess the long-term effects of gap junction inhibitors in intracerebral hemorrhage models. PMID- 19298759 TI - The effects of nicotinamide adenine dinucleotide on intracerebral hemorrhage induced brain injury in mice. AB - In the present study, we investigated whether the administration of nicotinamide adenine dinucleotide (NAD+) provides brain protection in a mouse model of intracerebral hemorrhage (ICH). Male CD-1 mice were divided into sham, ICH treated with vehicle and ICH treated with NAD+ (10 or 20 mg/kg, intranasal) groups. Intranasal delivery of NAD+ resulted in an increase in NAD+ contents in the brains. ICH was induced by collagenase injection. Neurological function, hemorrhage volume and brain edema were measured 24 hours after injection. ICH caused significant neurological deficit with associated brain edema. NAD+ (10 and 20 mg/kg) failed to reduce brain injury after ICH. These results suggest that NAD+ has no neuroprotective effect at 24 hours after ICH. PMID- 19298760 TI - Albumin ameliorates tissue plasminogen activator-mediated blood-brain barrier permeability and ischemic brain injury in rats. AB - OBJECTIVE: Tissue plasminogen activator (tPA) may aggravate ischemic neuronal damage after focal cerebral ischemia and increase blood-brain barrier permeability. Human serum albumin has neuroprotective effects on ischemic stroke. However, whether albumin can attenuate the deleterious effects of tPA is yet unknown. METHODS: In the present study, we attempted to determine the effects of albumin on cerebral injury and blood-brain barrier disruption induced by middle cerebral artery occlusion for 2 hours followed by 24 hours of reperfusion and tPA. RESULTS: We found that infarct volume in rats which received saline and tPA was 35.6 +/- 3.8% (mean +/- SEM) and 50.9 +/- 4.5%, respectively. There was significant difference between the two groups (p<0.01). The infarct volume in rats that received tPA with albumin was significantly decreased to 29.2 +/- 3.3% (p<0.05), compared with tPA-only-treated group. Combination therapy using tPA with albumin also improved neurological deficits and reduced brain edema significantly (p<0.05). Relative to tPA-treated group, rats that received combination therapy using tPA with albumin had significantly reduced blood-brain barrier permeability, evaluated by quantitation of Evans blue leakage (p<0.05). CONCLUSION: In acute ischemic stroke, combination therapy using tPA with albumin can attenuate the deleterious effects of tPA. PMID- 19298761 TI - Glutathione peroxidase and subarachnoid hemorrhage: implications for the role of oxidative stress in cerebral vasospasm. AB - OBJECTIVE: Worldwide, cerebral vasospasm after subarachnoid hemorrhage (SAH) has an estimated morbidity and mortality of 1.2 million annually. While it has long been suspected that reactive oxygen species play a major role in the etiology of cerebral vasospasm after SAH, promising results in animal work were not borne out in human clinical trials, despite intensive research effort. The purpose of this study is to investigate the role of glutathione peroxidase in the SAH cerebrospinal fluid milieu. METHODS: We utilized commercially available kits for the quantitation of glutathione peroxidase 1 (glutathione peroxidase) activity and oxygen radical capacity and sodium dodecyl sulfate polyacrylamide gel electrophoresis with Western blotting with specific antibodies to human glutathione peroxidase to determine the enzyme content of the cerebrospinal fluid samples. Human cerebrospinal fluid was obtained in an Institutional Review Board exempt manner for this study in the following groups: control (no SAH), CSF(C) (SAH but no vasospasm on angiography) and CSF(V) (SAH with clinical and angiographic vasospasm). RESULTS: We found that glutathione peroxidase activity is significantly higher in CSF(V) compared with CSF(C), and this is reflected in a higher total oxidative capacity in CSF(V). Despite similar levels of glutathione peroxidase protein, CSF(V) had significantly greater activity than CSF(C). DISCUSSION: These results further elucidate previous research from this laboratory, showing increased oxidative stress in CSF(V) compared with CSF(C). In conclusion, there appears to be increased glutathione peroxidase activity in CSF(V), despite there being increased levels of oxidative stress markers, suggesting overwhelming oxidative stress may play a role in cerebral vasospasm after SAH. PMID- 19298762 TI - Neurogenesis-related genes expression profiling of mouse fibroblastic stem cells induced by Wnt signaling. AB - OBJECTIVE: Wnt signaling has been implicated in the control of cell growth and differentiation during the development of the central nervous system. In this work, we assessed the role of Wnt pathway in the regulation of genes related to the neuronal differentiation of stem cells. METHODS: C3H10T1/2 mouse mesenchymal stem cells were transfected with the Ad5 recombinant plasmid encoding Wnt3A. The Wnt3A-induced gene expression profile of these cells was determined with the Affymetrix MG430A 2.0 chip. The microarray results were confirmed by real-time polymerase chain reaction. RESULTS: Several differentially expressed genes were identified in the Wnt3A-transfected cells. Many of these genes are targets of the Wnt signaling pathway, such as those encoding beta-catenin, Frizzled7, Frizzled1, LRP5, Dishevelled, axin, and GSK-3beta. Transfection of Wnt modulated the expression of genes of neuronal differentiation of these stem cells. CONCLUSION: This work demonstrated that Wnt is an important regulator gene for the expression of neurogenesis of stem cells. PMID- 19298763 TI - The effects of blood pressure and urokinase on brain injuries after experimental cerebral infarction in rats. AB - OBJECTIVE: With the proposal of penumbra theory and development of intra-arterial thrombolysis (such as urokinase), the outcome of ischemic cerebrovascular disease is greatly improved. However, the incidence of hemorrhagic transformation (HT) increased concomitantly, and some studies showed a close relationship between blood pressure and HT. The mechanisms of blood pressure and urokinase effect on the incidence of HT are not clear. In this study, we investigated the effects of the different levels of blood pressure and urokinase on the ischemic lesions, the incidence of HT and the expression of matrix metalloproteinase 9 (MMP-9) in the rat ischemia-reperfusion models. METHODS: Temporary focal ischemia was induced in male Sprague-Dawley rats using the intraluminal vascular occlusion method. The animals were assigned into four groups (n=11 in each group): low blood pressure group (LP), normal blood pressure group (NP), high blood pressure group (HP) and urokinase/high blood pressure group (UKHP). Adnephrin was applied to enhance the mean arterial blood pressure (MABP) at the beginning of reperfusion. MABP was maintained 20 mmHg higher than the baseline for 1 hour. Sodium nitroprusside was used to decrease MABP by 20 mmHg lower than the baseline for 1 hour. Both urokinase and adnephrin were used concomitantly in the UKHP group. Neurological deficit scores were evaluated at 2 hours (R2h) and 24 hours (R24h) after reperfusion. All rats were decapitated, their brains were sliced into 15-mum thick slices, and the infarct volume and the visible HT were analysed. Three rats in each group were taken for immunohistochemistry and pathological analysis. RESULTS: There was no significant difference in MABP among the groups at the baseline time points (p>0.05), but blood pressure are definitely increased and decreased in the HP, UKHP, and LP groups. Neurological deficit scores showed no significant difference at R2h among the groups (p=0.443). However, neurological deficit scores showed significant differences at R24h among the groups, the neurological deficits scores of rats in the LP group are significantly higher than that in the other groups. Compared with that of 2 hours after reperfusion, neurological deficit scores deteriorated in the LP group (p=0.047) but was improved in the NP, HP and UKHP groups (p=0.076, 0.002, 0.017, respectively) at 24 hours after reperfusion. The infarct volume in the HP group was apparently smaller than that in the LP group (p=0.006). There was indeed a tendency that HT occurred more frequently in the UKHP group (42.8%) than in the HP (25%) and LP (28.5%) groups. MMP-9 expression showed significant increase around the ischemic lesion areas of the UKHP group and significant decrease in the cortical areas of the LP and HP groups but no significant difference in the basal ganglia of rats of all groups. CONCLUSION: Mild elevation of blood pressure during reperfusion is supposed to improve neurological outcomes in rats following ischemia/reperfusion. The incidence of HT tended to increase with the elevation of blood pressure and the administration of urokinase. Immunohistochemitry analysis indicated that incidence of HT may correlate with excessive expression of MMP-9. PMID- 19298764 TI - Hemorrhagic profile of the fibrinolytic alfimeprase after ischemia and reperfusion. AB - OBJECTIVE: Recanalization therapies for ischemic stroke have been slow to change clinical practice because of perceived and published risks of hemorrhage associated with lytic administration. We quantified alfimeprase in an acute ischemia-reperfusion model, as compared with recombinant tissue plasminogen activator, with hemorrhagic transformation as the primary endpoint and infarction volume and blood-brain barrier permeability as secondary endpoints. METHODS: Five groups were studied in a blinded fashion: alfimeprase at doses of 0.03 (n=8), 0.1 (n=11) and 0.3 mg/kg (n=8); recombinant tissue plasminogen activator at 1 mg/kg (n=9); carrier infused controls (n=9). The middle cerebral artery was occluded for 5 hours followed by removal of the suture for reperfusion. Drugs were infused immediately following reperfusion over a 10-minute period. Approximately 24 hours later, the animals were anesthetized and decapitated, and the brains were rapidly harvested and frozen. Serial brain sections were obtained and inspected for hemorrhages. Infarction and blood-brain barrier permeability were also evaluated in additional experiments in control, 0.1 mg/kg alfimeprase and 1 mg/kg recombinant tissue plasminogen activator-treated rats. RESULTS: The hemorrhagic transformation frequency, neurological deficit and the mortality rate of alfimeprase were significantly lower than for recombinant tissue plasminogen activator at the 0.03 mg/kg dose and not statistically different at the higher doses. Infarction and blood-brain barrier permeability were not significantly different among control, 0.1 mg/kg alfimeprase and recombinant tissue plasminogen activator. DISCUSSION: In this model, alfimeprase, a new fibrinolytic agent, exhibits a profile comparable to recombinant tissue plasminogen activator. PMID- 19298765 TI - Nail psoriasis and biologics. AB - BACKGROUND: Since the advent of biologic therapies for psoriasis, reports of efficacy in nail psoriasis have appeared in the literature and at international conferences with increasing frequency. OBJECTIVE: This article aims to review the existing literature on the use of biologics in the treatment of nail psoriasis. METHODS: An extensive literature review was conducted using OVID Medline. Studies examining the efficacy of biologics in the treatment of nail psoriasis were documented. RESULTS: A literature review revealed few clinical trials specifically concentrating on nail psoriasis; however, nails have been assessed in larger clinical trials for cutaneous psoriasis. A large, multicenter, phase III, double-blind, placebo-controlled study of infliximab administered as a brief induction regimen at weeks 0, 2, and 6 followed by a single infusion every 8 weeks revealed statistically significant mean percent improvement in the Nail Psoriasis Severity Index (NAPSI) score over placebo at both week 10 (26.8% vs 7.7%, respectively; p < .001) and week 24 (57.2% vs -4.1%, respectively; p < .001). For other biologics, evidence has thus far been largely anecdotal, appearing as either case studies or extracted secondarily from open-label prospective trials in plaque psoriasis or psoriatic arthritis. CONCLUSION: Infliximab appears to be the most effective treatment for nail psoriasis to date. PMID- 19298766 TI - Use of biologic therapeutics in difficult-to-treat psoriasis. AB - BACKGROUND: A need for improved psoriasis management drove the development of four biologicals now approved for use in moderate to severe psoriasis: alefacept, efalizumab, etanercept, and infliximab. OBJECTIVE: This review compares the recommended and practical use of biologicals in "difficult-to-treat" psoriasis. Difficult-to-treat patients, a subpopulation of particular interest to caregivers and regulators, are those for whom conventional options are unavailable, contraindicated, intolerable, or ineffective. CONCLUSIONS: Despite guidance recommending that biologicals be considered among first-line antipsoriasis treatments, formulary inclusion may be influenced by the biologicals' success in controlling difficult-to-treat psoriasis. Findings from the Clinical Experience Acquired with Raptiva (CLEAR) randomized controlled trial (RCT) established efalizumab's equal efficacy in difficult-to-treat and moderate to severe psoriasis. The CLEAR data were cited in support of a regulatory recommendation to include efalizumab in Canadian provincial formularies. For some other biologicals, evidence regarding efficacy in difficult-to-treat psoriasis remains equivocal. Additional RCTs are needed to define appropriate roles for specific biologicals in difficult-to-treat patients. PMID- 19298767 TI - Results of radiation therapy for treatment of classic Kaposi sarcoma. AB - BACKGROUND: Classic Kaposi sarcoma (CKS) is a vascular neoplasm that primarily affects men of Mediterranean and Ashkenazi Jewish descent. A variety of therapeutic options exist, and choice of treatment depends on clinical form and stage, as well as lesion location and size; options include surgical excision, intralesional interferon alpha-2b, local or extended field radiotherapy, and chemotherapy. OBJECTIVE: The aim of this study was to review the outcome of radiation therapy in the treatment of CKS at a single institution. METHODS: This retrospective study reviewed patients who receive radiation therapy for histologically confirmed CKS between 1994 and 2006. RESULTS: Sixteen patients were reviewed; the mean age at diagnosis was 74 years, and 13 patients were male. Fifteen patients (94%) presented with leg lesions, and two patients (12.5%) presented with arm lesions. The most commonly prescribed radiation dose was 30 Gy in 15 daily fractions of 2 Gy. All lesions responded to treatment, with a complete response rate of 88% and a partial response rate of 12%. Toxicity was limited to grade I dermatitis (four patients) and grade II dermatitis (two patients). CONCLUSION: Radiation therapy is an effective treatment modality for CKS and is associated with minimal toxicity. PMID- 19298768 TI - Tanning behavior of London-area youth. AB - BACKGROUND: The World Health Organization recommends that tanning parlors not be used by individuals under age 18 years. The impact of self-tanners on tanning parlor and sunscreen use by Canadian teens is unknown. OBJECTIVE: To determine teens' use of, knowledge of, attitudes toward, and behavior regarding tanning parlors, self-tanning products, and sunscreen. METHODS: Self-report questionnaire of grade 10 students in Thames Valley district, Ontario. RESULTS: Indoor tanning parlors were used by 14%, self-tanners by 28%, and sunscreen daily or most days by 36%. Use was more common in females. There was a strong association between parental and child use of tanning parlors and use of self-tanners. Sixty-two percent believe that self-tanners give adequate photoprotection. CONCLUSIONS: Further education is required to address teen tanning parlor use, infrequent sunscreen use, and lack of knowledge about the safety of tanning parlors and self tanning products. PMID- 19298769 TI - Surgical management of chest wall tuberculosis. AB - OBJECTIVE: Tuberculosis remains one of the most fatal diseases in the world. Extrapulmonary tuberculosis increases in parallel fashion with the rise of incidence of pulmonary tuberculosis. Tuberculosis of the chest wall is rare. METHODS: Our study retrospectively evaluated 10 patients with tuberculosis of the chest wall who were treated with surgery in addition to antituberculous chemotherapy in the last 8 years. Seven of the lesions were located on the anterior chest wall, and three of the lesions were on the lateral chest wall. RESULTS: Surgical therapy consisted of evacuation of the cold abscess material from the cavities and wide debridement of the soft tissue planes in all patients. Partial rib resections were performed in six patients owing to costal or chondral involvement. CONCLUSIONS: Chest wall tuberculosis must be considered in the differential diagnosis of chest wall lesions, especially in endemic areas. PMID- 19298770 TI - Occurrence of plantar pustular psoriasis during treatment with infliximab. AB - BACKGROUND: Pustular psoriasis is an uncommon form of psoriasis that often affects areas of the hands and feet. It typically manifests as small pustules that develop within erythematous areas of the palms and soles. Infliximab, a tumor necrosis factor inhibitor, can be used to treat pustular psoriasis. Infliximab can also be effective in the treatment of various other disorders, including plaque-type psoriasis, psoriatic arthritis, Crohn disease, rheumatoid arthritis, and ankylosing spondylitis. OBJECTIVE: We present a case of a young woman developing pustular psoriasis for the first time despite being on infliximab treatment for Crohn disease. RESULTS: Infliximab has been successful in the treatment of pustular psoriasis. In rare cases, plaque psoriasis appears for the first time during infliximab treatment for other disorders, such as Crohn disease. CONCLUSION: Plantar pustular psoriasis occurring for the first time during infliximab treatment is an uncommon occurrence. PMID- 19298771 TI - Acral, pure cutaneous, self-healing, late-onset, cellulitis-like Langerhans cell histiocytosis. AB - BACKGROUND: An unusual and not yet reported clinical presentation of pure cutaneous Langerhans cell histiocytosis (LCH) that was a diagnostic dilemma. OBJECTIVE: To bring to light the newer presentation of this disease. METHOD: The case was managed on clinical grounds initially as cellulitis. However, after the investigations and confirmation of the diagnosis, she was managed symptomatically. RESULTS: Pure cutaneous LCH may manifest in early adulthood with features of cellulitis, erythematous edematous vesicular plaques, and nodules distributed in the flexural acral areas. CONCLUSION: Pure cutaneous LCH may have an acute late onset with unusual morphology as cellulitis, inverse acral distribution, and self-healing. PMID- 19298772 TI - Alopecia areata following adalimumab. AB - BACKGROUND: Alopecia areata is a nonscarring hair loss characterized by well circumscribed patchy areas, most often on the scalp. The inflammatory cytokine tumor necrosis factor alpha (TNF-alpha), has been connected with the development of alopecia areata in vivo; thus, the TNF-alpha inhibitors have been cited as possible treatments for this autoimmune condition. OBJECTIVE: We report a case of alopecia areata that developed in a 52-year-old woman who was recently started on adalimumab for treatment of her psoriatic arthritis. RESULTS: We discuss the previously published cases in the literature linking alopecia areata to TNF-alpha inhibitor administration. Our case is the first report of a new-onset alopecia areata following adalimumab. CONCLUSIONS: Even though TNF-alpha is implicated in causing alopecia areata, TNF-alpha inhibitors have paradoxically been associated with new cases of alopecia areata. It is possible that TNF-alpha may not be involved in the pathogenesis of alopecia areata, as in vitro studies have suggested. PMID- 19298773 TI - Anticoagulant therapy: should it be discontinued before cutaneous surgery? PMID- 19298774 TI - Celiac disease. PMID- 19298775 TI - Clinical efficacy of 25% L-ascorbic acid (C'ensil) in the treatment of melasma. AB - BACKGROUND: L-Ascorbic acid is used to treat melasma; however, it is quickly oxidized in aqueous solutions. Thus, C'ensil, a formulation containing 25% l ascorbic acid and a chemical penetration enhancer, was created to promote the penetration of l-ascorbic acid into the skin. OBJECTIVE: To evaluate the efficacy of C'ensil in patients with melasma. METHODS: Forty subjects with melasma were treated with C'ensil during an open-label trial over a period of 16 weeks. Each subject's skin pigmentation was assessed every 4 weeks using the Melasma Area and Severity Index (MASI) and mexameter score. In addition, transepidermal water loss, skin dryness and irritation, and quality of life (Melasma Quality of Life Scale [MelasQoL]) were evaluated. RESULTS: After 16 weeks, a significant decrease was noted in the degree of pigmentation based on the patients' MASI and mexameter scores. MelasQoL scores also decreased, indicating an increase in the subjects' quality of life. CONCLUSION: Our data indicate that C'ensil is an effective treatment modality for melasma. PMID- 19298776 TI - Prevalence of non-organ-specific autoantibodies in patients with pemphigus vulgaris. AB - BACKGROUND: Pemphigus vulgaris (PV) is a potentially life-threatening, organ specific, autoimmune, blistering disease of the skin and mucous membranes. Although several reports suggest an association between pemphigus and other autoimmune connective tissue disorders, studies that measure non-organ-specific autoantibodies are lacking. OBJECTIVE: To evaluate the prevalence of antinuclear antibodies (ANA), anti-double-stranded DNA (anti-dsDNA) antibodies, and antibodies against extractable nuclear antigens (ENAs) in PV patients. METHODS: Serum samples were obtained from 59 PV patients and 50 healthy controls. Indirect immunofluorescence assays containing human epithelial cell substrates (HEp-2) and Crithidia luciliae were used to detect ANA and anti-dsDNA antibodies, respectively. A multiplexed addressable laser bead immunoassay was employed to measure autoantibodies to: Smith (Sm), ribonucleoprotein (RNP), Sjogren syndrome B (SSB/La), Sjogren syndrome A (SSA/Ro), histidyl transfer ribonucleic acid synthetase (Jo-1), topoisomerase I (Scl-70), and ribosome-P (Ribo-P) antigens. RESULTS: Positive ANAs were obtained in 22 of 59 (37.3%) PV patients compared with 4 of 50 (8.0%) healthy controls (p or =75 years of age. Older patients had more cardiovascular risk factors and had a higher acuity at presentation. Patients age > or =75 years treated with bivalirudin alone had similar ischemic outcomes, but significantly lower rates of bleeding compared with those treated with heparin and glycoprotein IIb/IIIa inhibitors overall and in the PCI subset. The number needed to treat with bivalirudin alone to avoid 1 major bleeding event was lower in this age group (23 overall and 16 for PCI-treated patients) than in any other. CONCLUSIONS: Ischemic and bleeding complications after NSTE-ACS increase with age. Although ischemic event rates are not statistically different with either bivalirudin alone or a heparin plus glycoprotein IIb/IIIa inhibitor, bleeding complications are significantly less frequent with bivalirudin alone. Because of the substantial risk of bleeding in patients age > or =75 years, the number needed to treat to avoid 1 major bleeding event using bivalirudin alone was the lowest in the elderly group, especially among those undergoing PCI. PMID- 19298915 TI - 9-month clinical, angiographic, and intravascular ultrasound results of a prospective evaluation of the Axxess self-expanding biolimus A9-eluting stent in coronary bifurcation lesions: the DIVERGE (Drug-Eluting Stent Intervention for Treating Side Branches Effectively) study. AB - OBJECTIVES: This study sought to assess the safety and performance of the Axxess (Devax Inc., Lake Forest, California) self-expanding drug-eluting stent in coronary bifurcation lesions. BACKGROUND: Percutaneous treatment of coronary bifurcations is a predictor of adverse late outcomes, in part because of the lack of dedicated devices. METHODS: Patients with de novo bifurcation lesions were prospectively enrolled in a multicenter study. The Axxess stent was deployed at the level of the carina followed by additional sirolimus-eluting stents in the distal parent vessel (PV) and/or side branch (SB). All patients underwent clinical follow-up at 9 months; 150 were to receive control angiography and 76 were to receive intravascular ultrasound. The primary end point was the rate of major adverse cardiac events (MACE): a composite of death, myocardial infarction (MI), and target lesion revascularization (TLR). Secondary end points included in segment restenosis, late loss, and percent neointimal volume obstruction. RESULTS: Overall, 302 patients were treated with 299 Axxess stents (99%). Additional stenting of 1 branch was performed in 21.7% of patients (17.7% PV, 4% SB), and of both branches in 64.7%. At 9 months, 99.3% of patients returned for clinical follow-up; from the angiographic and IVUS substudies, 93.3% and 89.4% returned. The cumulative 9-month MACE rate was 7.7% (0.7% death, 3.3% non-Q-wave MI, 1.0% Q-wave MI, 4.3% TLR). Subacute and late stent thrombosis occurred in 0.7% and 0.3% of patients. Total restenosis was 6.4% (3.6% PV, 4.3% SB), late loss was 0.20 +/- 0.41 mm in the PV and 0.17 +/- 0.34 mm in the SB. In the Axxess stent segment, percent neointimal volume obstruction was 4.3 +/- 5.2%. CONCLUSIONS: This prospective multicenter study confirms the safety and performance of the Axxess stent in bifurcation lesions. (Drug-Eluting Stent Intervention for Treating Side Branches Effectively; ACTRN12606000259549). PMID- 19298916 TI - Renal protective effects and the prevention of contrast-induced nephropathy by atrial natriuretic peptide. AB - OBJECTIVES: This study was designed to examine the protective effects of atrial natriuretic peptide (ANP) on contrast-induced nephropathy (CIN) after coronary angiography. BACKGROUND: Contrast-induced nephropathy is a common complication after angiography. Some studies have shown that ANP has renal protective effects, but the beneficial effects for CIN prevention remain to be clearly shown. METHODS: In a prospective, controlled, randomized trial in 254 consecutive patients with serum creatinine concentrations of > or =1.3 mg/dl, patients received either ANP (0.042 microg/kg/min; ANP group, n = 126) or Ringer solution alone (control group, n = 128). Treatment of either type was initiated 4 to 6 h before angiography and continued for 48 h. RESULTS: There were no significant differences in age, sex, diabetes mellitus, or baseline serum creatinine level between the 2 groups. The prevalence of CIN, defined as a 25% increase in creatinine or an increase in creatinine of > or =0.5 mg/dl from baseline within 48 h, was significantly lower in the ANP group than in the control group (3.2% vs. 11.7%, respectively; p = 0.015). Multivariate analysis revealed that the use of >155 ml of contrast medium (odds ratio: 6.89; p < 0.001) and ANP treatment (odds ratio: 0.24; p = 0.016) were significant predictors of developing CIN. The incidence of an increase in creatinine of > or =25% or of > or =0.5 mg/dl from baseline at 1 month was also significantly lower in the ANP group than in the control group (p = 0.006). CONCLUSIONS: In addition to hydration, ANP administration is effective in the prevention of CIN in patients with chronic renal failure, and the effect was maintained for 1 month. PMID- 19298917 TI - Atrial natriuretic peptide for the prevention of contrast-induced nephropathy: what's old is new but at the right dose and duration of therapy! PMID- 19298918 TI - The utility of 12-lead Holter monitoring in patients with permanent atrial fibrillation for the identification of nonresponders after cardiac resynchronization therapy. AB - OBJECTIVES: This study sought to determine the incidence of ineffective capture using 12-lead Holter monitoring and to assess whether this affects response to cardiac resynchronization therapy (CRT). BACKGROUND: Cardiac resynchronization therapy is used in patients with atrial fibrillation (AF), prolonged QRS duration, and heart failure in the setting of ventricular dysfunction. The percentage of ventricular pacing is used as an indicator of adequate biventricular (BiV) pacing. Although device counters show a high pacing percentage, there may be ineffective capture because of underlying fusion and pseudo-fusion beats. METHODS: We identified 19 patients (age 72 +/- 8 years, ejection fraction 18 +/- 5%), with permanent AF who underwent CRT. All patients received digoxin, beta-blockers, and amiodarone for rate control; device interrogation showed >90% BiV pacing. Patients had a 12-lead Holter monitor to assess the presence of effective (>90% fully paced beats/24 h) pacing. At 12 months post-CRT, the New York Heart Association functional class was reassessed and an echocardiogram was obtained and compared with pre-CRT. RESULTS: Only 9 (47%) patients had effective pacing. The other 10 (53%) patients had 16.4 +/- 4.6% fusion and 23.5 +/- 8.7% pseudo-fusion beats. Long-term responders (> or =1 New York Heart Association functional class improvement) to CRT had a significantly higher percentage of fully paced beats (86.4 +/- 17.1% vs. 66.8 +/- 19.1%; p = 0.03) than nonresponders. CONCLUSIONS: Pacing counters overestimate the degree of effective BiV pacing in patients with permanent AF undergoing CRT therapy. Only patients with complete capture responded clinically to CRT. These findings have important implications for the application of CRT to patients with permanent AF and heart failure. PMID- 19298919 TI - Associations of borderline and low normal ankle-brachial index values with functional decline at 5-year follow-up: the WALCS (Walking and Leg Circulation Study). AB - OBJECTIVES: We studied associations of borderline and low normal ankle-brachial index (ABI) values with functional decline over a 5-year follow-up. BACKGROUND: The associations of borderline and low normal ABI with functional decline are unknown. METHODS: The 666 participants included 412 with peripheral arterial disease (PAD). Participants were categorized as follows: severe PAD (ABI <0.50), moderate PAD (ABI 0.50 to 0.69), mild PAD (ABI 0.70 to 0.89), borderline ABI (0.90 to 0.99), low normal ABI (1.00 to 1.09), and normal ABI (ABI 1.10 to 1.30). Outcomes were assessed annually for 5 years. Mobility loss was defined as loss of the ability to walk one-quarter mile or walk up and down 1 flight of stairs without assistance among participants without baseline mobility impairment. Becoming unable to walk for 6 min continuously was defined as stopping during the 6-min walk at follow-up among those who walked for 6 min continuously at baseline. Results were adjusted for age, sex, race, comorbidities, and other confounders. RESULTS: Hazard ratios (HRs) for mobility loss according to ABI category were as follows: severe PAD, HR: 4.16 (95% confidence interval [CI]: 1.58 to 10.92); moderate PAD, HR: 3.82 (95% CI: 1.66 to 8.81); mild PAD, HR: 3.22 (95% CI: 1.43 to 7.21); borderline ABI, HR: 3.07 (95% CI: 1.21 to 7.84); and low normal ABI, HR: 2.61 (95% CI: 1.08 to 6.32; p trend = 0.0018). Similar associations were observed for becoming unable to walk for 6 min continuously (p trend < 0.0001). CONCLUSIONS: At 5-year follow-up, persons with borderline ABI values have a higher incidence of mobility loss and becoming unable to walk for 6 min continuously compared with persons who have a normal baseline ABI. A low normal ABI is associated with an increased incidence of mobility loss compared with persons who have a normal ABI. PMID- 19298920 TI - Rethinking the morbidity of peripheral arterial disease and the "normal" ankle brachial index. PMID- 19298921 TI - Hypoplastic left heart syndrome links to chromosomes 10q and 6q and is genetically related to bicuspid aortic valve. AB - OBJECTIVES: This study was designed to identify disease loci for hypoplastic left heart syndrome (HLHS) and evaluate the genetic relationship between HLHS and bicuspid aortic valve (BAV). BACKGROUND: Previously, we identified that HLHS and BAV exhibit complex inheritance, and both HLHS and BAV kindreds are enriched for BAV. However, the genetic basis of HLHS and its relationship to BAV remains unclear. METHODS: Family-based nonparametric genome-wide linkage analysis was performed in kindreds ascertained by either an HLHS or BAV proband. Echocardiograms were performed on 1,013 participants using a sequential sampling strategy (33 HLHS kindreds, 102 BAV kindreds). RESULTS: The recurrence risk ratio of BAV in HLHS families (8.05) was nearly identical to that in BAV families (8.77). Linkage to chromosomal regions 10q22 and 6q23 with maximum logarithm of odds scores of 3.2 and 3.1, respectively, was identified in HLHS kindreds. In addition, 5 suggestive loci were identified (7q31, 11q22, 12q13, 14q23, and 20q11). We previously identified loci at chromosomes 18q22, 13q34, and 5q21 in BAV kindreds. The relationship between these loci was examined in the combined HLHS and BAV cohort and associations between loci were demonstrated (5q21, 13q34, and 14q23; 6q23 and 10q22; 7q31 and 20q11). Subsequent subsets linkage analysis showed a significant improvement in the logarithm of odds score at 14q23 only (4.1, p < 0.0001). CONCLUSIONS: These studies demonstrate linkage to multiple loci identifying HLHS as genetically heterogeneous. Subsets linkage analyses and recurrence risk ratios in a combined cohort provide evidence that some HLHS and BAV are genetically related. PMID- 19298922 TI - Hypoplastic left heart syndrome: new genetic insights. PMID- 19298923 TI - Ambulatory monitoring of congestive heart failure by multiple bioelectric impedance vectors. AB - OBJECTIVES: This study was designed to investigate the properties of multiple bioelectric impedance signals recorded during congestive heart failure (CHF) by utilizing various electrode configurations of an implanted cardiac resynchronization therapy system. BACKGROUND: The monitoring of CHF has relied mainly on right-side heart sensors. METHODS: Fifteen normal dogs underwent implantation of cardiac resynchronization therapy systems using standard leads. An additional left atrial (LA) pressure lead sensor was implanted in 5 dogs. Continuous rapid right ventricular (RV) pacing was applied over several weeks. Left ventricular (LV) catheterization and echocardiography were performed biweekly. Six steady-state impedance signals, utilizing intrathoracic and intracardiac vectors, were measured through ring (r), coil (c), and device Can electrodes. RESULTS: Congestive heart failure developed in all animals after 2 to 4 weeks of pacing. Impedance diminished gradually during CHF induction, but at varying rates for different vectors. Impedance during CHF decreased significantly in all measured vectors: LV(r)-Can, -17%; LV(r)-RV(r), -15%; LV(r)-RA(r), -11%; RV(r)-Can, -12%; RV(c)-Can, -7%; and RA(r)-Can, -5%. The LV(r)-Can vector reflected both the fastest and largest change in impedance in comparison with vectors employing only right-side heart electrodes, and was highly reflective of changes in LV end-diastolic volume and LA pressure. CONCLUSIONS: Impedance, acquired by different lead electrodes, has variable responses to CHF. Impedance vectors employing an LV lead are highly responsive to physiologic changes during CHF. Measuring multiple impedance signals could be useful for optimizing ambulatory monitoring in heart failure patients. PMID- 19298924 TI - The cardiovascular programs of the National Heart, Lung, and Blood Institute: from vision to action to impact. PMID- 19298925 TI - Comparative effectiveness research: the view from the NHLBI. PMID- 19298926 TI - Multiple congenital left ventricular diverticula and aneurysm. PMID- 19298927 TI - Management of acute limb ischemia: current perspectives. Introduction. PMID- 19298928 TI - Historical reflections on the management of acute limb ischemia. AB - The history of clot removal consists primarily of efforts at procedures, techniques, and instruments that had suboptimal performance. In the 1960s, the amputation and death rates following attempts at removal were as high as 50%. A wide variety of instruments were used but none were specifically designed for embolus and thrombus removal. The need was obvious but the problem was unresolved. PMID- 19298929 TI - Clinical staging of acute limb ischemia as the basis for choice of revascularization method: when and how to intervene. AB - In acute lower limb ischemia, there are basically three management options: (1) clot removal by catheter-directed thrombolysis with or without percutaneous mechanical thrombectomy, (2) surgical thromboembolectomy followed by correction of underlying arterial lesions, and (3) anticoagulation with continued observation. Arterial embolic occlusion presents more abruptly and with more severe ischemia than arterial thrombosis, which occurs in narrowed arterial segments that have generally developed some degree of collateral circulation. The appropriate choice of treatment for acute limb ischemia depends to a great extent on the severity of the ischemia. Level of ischemia is readily determined by examining for sensory loss or motor deficit and interrogating the distal arteries and veins for audible flow signals with a handheld Doppler velocity detector. After clot removal, appropriate management of the responsible underlying lesion depends on its characteristics, best determined by vascular imaging. Staging the severity of ischemia according to clinical classification levels in the current reporting standards for lower extremity ischemia continues to serve as the basis for logical management decisions. This approach is outlined in algorithmic form and alternative pathways are discussed in this article. PMID- 19298930 TI - Proper evaluation and management of acute embolic versus thrombotic limb ischemia. AB - Acute limb ischemia is one of the most potentially devastating but treatable diseases faced by the vascular surgeon. It is crucial to identify the ischemic process early, as the outcomes of early intervention can lead to limb salvage, whereas late recognition places the patient at risk for limb loss and potential mortality. This article emphasizes the importance of properly evaluating the acutely ischemic extremity to ascertain whether the etiology is embolic or thrombotic. Appropriate treatment and management of the limb are determined by the underlying cause of the ischemia. The authors will review the clinical features, angiographic findings, and strategies for management for these similar but distinct etiologies: acute embolic versus thrombotic limb ischemia. PMID- 19298931 TI - Acute limb ischemia due to popliteal artery aneurysm: a continuing surgical challenge. AB - Up to 50% of all popliteal artery aneurysms (PAA) present with acute limb ischemia (ALI). ALI due to PAA is a difficult surgical problem, with a 20% to 60% incidence of limb loss and up to 12% mortality reported in the literature in the last three decades. Imminent limb threat requires emergency infrainguinal reconstruction, preferably with autogenous conduit. ALI due to PAA is limb threatening, often due to obliteration of the tibial vessels in addition to thrombosis of the PAA itself. Arteriography is needed to define inflow vessel and outflow vessel anatomy followed by thrombectomy of the run-off vasculature to establish an appropriate target for bypass. Patients without evidence of neurologic deficit are best served by formal arteriography. Intraarterial thrombolysis is used to establish an outflow vessel for bypass if no runoff vessels are visible. In general, emergency operations are associated with inferior patency and limb salvage compared to elective procedures. Endovascular exclusion of PAA with covered stent graft is used increasingly in the elective setting and has been reported in patients presenting with limb ischemia. The following discussion outlines our algorithm in managing ALI from PAA and reviews management decisions and results of treatment. PMID- 19298932 TI - Acute arterial occlusion secondary to trauma. AB - During the last century, the treatment paradigm of extremity vascular injuries has evolved from primary ligation to the current application of endovascular techniques. Regardless of the method employed, management of traumatic vascular injuries continues to pose a challenge to the treating surgeon, especially when combined with soft tissue and bony injuries. This article reviews the current trends in diagnosis and treatment of acute arterial occlusions secondary to vascular trauma. PMID- 19298933 TI - Metabolic consequences of acute limb ischemia and their clinical implications. AB - Acute limb ischemia is a common medical condition resulting from arterial embolization, in situ thrombosis, trauma, and other causes. The severity of injury is related to the duration of ischemia and the effects of reperfusion. Metabolic consequences of reperfusion injury can be variable, ranging from transient symptoms in the lower extremity to systemic inflammation with multiple organ dysfunction. This article provides an overview of some of the key mediators of reperfusion injury. Additional discussion is focused on the clinical effects of reperfusion in the extremity, as well as the pulmonary, cardiac, and renal organ systems. A better understanding of these processes may result in improved patient outcomes and decreased mortality. PMID- 19298934 TI - Contemporary management of acute limb ischemia: factors associated with amputation and in-hospital mortality. AB - Contemporary treatment of patients with acute limb ischemia includes both open and endovascular techniques, and advances in technology continue to make interventions easier and safer. However, the greatest gains in improving outcomes in these patients will come from more consistent and prompt recognition of the disease followed by rapid, standardized therapy to minimize the risk of limb loss and subsequent reperfusion-related injury. This article uses case reports to highlight critical issues that may arise during the course of diagnosis and treatment and describes clinical factors associated with morbidity and mortality in these patients. PMID- 19298935 TI - Do randomized trials of thrombolysis versus open revascularization still apply to current management: what has changed? AB - Several randomized trials of catheter-directed thrombolysis versus operative revascularization in patients with acute lower extremity ischemia were performed in the mid-1990 s. Although the outcomes of these trials were not definitive, they did provide insight into potential uses and techniques of catheter-directed thrombolysis and lytic agents. This article reviews the outcomes of these randomized trials and describes advances in thrombolytic techniques and technology, including percutaneous mechanical thrombectomy devices and innovative catheter designs that accelerate lysis and the development of direct-acting lytic agents. PMID- 19298936 TI - Can intraoperative thrombolytic therapy assist with the management of acute limb ischemia? AB - Catheter-directed thrombolytic therapy is a well-accepted treatment modality for acute arterial and graft occlusion, but intraoperative use of plasminogen activators as an adjunct to operative thrombectomy or thromboembolectomy is frequently overlooked by vascular surgeons. Evidence exists that intraoperative intraarterial infusion of lytic agents is safe and is accompanied by breakdown of distal thrombus. This article describes the rationale for intraoperative thrombolysis, reviews the evidence supporting its use, and suggests several technical approaches to incorporating lytic therapy intraoperatively. PMID- 19298937 TI - Controlled reperfusion after acute and persistent limb ischemia. AB - Morbidity and mortality after revascularization of acute ischemic limbs remain high, despite many surgical improvements during the last decades. To a large extent, this is related to reperfusion itself after complete, acute, prolonged ischemia that results in postreperfusion syndrome. This report addresses the deleterious consequences of postreperfusion syndrome, which occurs after normal blood reperfusion, and describes the pathophysiologic basis for this complication. A unique opportunity arises from these data. Control of the initial reperfusion by modifying the conditions of reperfusion and the composition of the reperfusate will open the door to a new approach to salvage limbs and lives from this disastrous complication. The same approach that is used successfully in other organs and tissues (eg, heart, kidney, brain, whole body) will put the vascular surgeon in the position to intraoperatively counteract the consequences of revascularization after prolonged acute limb ischemia. PMID- 19298938 TI - The use of proliferation signal inhibitors in the prevention and treatment of allograft vasculopathy in heart transplantation. AB - Cardiac allograft vasculopathy (CAV) currently represents one of the most important causes of long-term morbidity and mortality in the heart transplant population. In well-designed studies with de novo patients, the use of proliferation signal inhibitors (PSIs; everolimus and sirolimus) has been shown to significantly prevent the intimal growth of graft coronary arteries in comparison to other immunosuppressive regimens, reducing the incidence of vasculopathy at 12 and 24 months. In addition, conversion to PSIs in maintenance patients with established CAV has also shown promising results in the reduction of the progression of the disease and its clinical consequences. For these reasons the interest shown by various transplantation units in the potential role of PSIs in this field is growing. The aim of the present article is to review the information obtained to date on the use of PSIs in heart transplant recipients, both in the prevention and the treatment of CAV. The principal published recommendations on the introduction and appropriate management of these drugs in clinical practice are also collected, as well as certain recommendations given by the authors based on their experience. PMID- 19298939 TI - Histocompatibility methods. AB - Predicting humoral alloimmune potential in transplant recipients is the objective of histocompatibility testing and depends upon accurate donor typing and sensitive and specific testing for antibodies to human leukocyte antigen. This review for the transplant clinician will describe the evolution and current practices of histocompatibility testing methods for typing, crossmatching and antibody screening. Novel methods of measuring T-cell alloimmune potential will also be briefly discussed. Emphasis is given to the clinical applicability and limitations of each test, and the collective consideration of all tests in concert, as part of the immunologic risk assessment of the solid organ transplant recipient. PMID- 19298940 TI - Evolving trends in induction therapy. AB - A decade of spectacular innovation in maintenance immunosuppressive drugs has resulted in dramatic reductions in acute rejection and improvement in short- and long-term outcomes after renal transplantation. However, the new drugs continue to lack specificity, many require frequent therapeutic drug monitoring, and all of them are associated with acute and chronic toxicities. The new biologic agents, monoclonal antibodies, and receptor-fusion proteins lack immunogenicity, have long half-life and prolonged biologic effects, require intermittent administration, and have minimal toxicity. The specificity and selectively of the targets of the new biologic agents render them less toxic than the oral maintenance drugs and thus could possibly replace those drugs most frequently associated with long-term toxicity such as the corticosteroids and the calcineurin inhibitors. PMID- 19298941 TI - Lung transplantation: does oxidative stress contribute to the development of bronchiolitis obliterans syndrome? AB - Lung transplantation is the ultimate treatment of end-stage lung disease. After transplantation, the 1-year survival rate is 80%. However, 5-year survival rates drop to 50% due to bronchiolitis obliterans syndrome (BOS). Ischemia/reperfusion injury, infections, and acute rejection are major risk factors contributing to the development of BOS. These risk factors are also associated with increased oxidative stress. Oxidative stress is a condition whereby prooxidants overwhelm the antioxidant defense system and may contribute to the pathogenesis of BOS by inducing more tissue injury and inflammation. This article reviews the current state of knowledge on oxidative stress in lung transplantation and BOS. PMID- 19298942 TI - Current indications for combined liver and kidney transplantation in adults. AB - A significant number of patients awaiting liver transplantation have associated renal failure. Combined Liver and Kidney Transplantation (CLKT) is increasingly offered especially since the introduction of Model for End-Stage Liver Disease (MELD). Decision to perform CLKT is straightforward when both organs suffer end stage failure. However, the indications for CLKT are not well defined and there is controversy concerning some. We reviewed available data on PUBMED, United Network for Organ Sharing (UNOS), Organ Procurement Transplantation Network (OPTN), European Society for Organ Transplantation (ESOT) and discuss all current indications for CLKT. CONCLUSION: Overall long-term outcome following CLKT is acceptable. There is an urgent need to further refine our ability to identify the cases with reversible renal injury in the setting of end-stage liver disease to avoid unnecessary CLKT. Liver protects the kidney from disease recurrence and allograft loss in metabolic diseases. However, the use of liver allograft for immunological protection of kidneys in highly sensitised patients with positive cross-match and previously failed renal transplants is still experimental. PMID- 19298943 TI - Biomarkers of the immunomodulatory effect of immunosuppressive drugs in transplant recipients. AB - Nowadays, monitoring of the immunosuppressive therapy received by solid organ transplant recipients is based on measuring drug blood levels to achieve concentrations within the established therapeutic range. This strategy largely prevents the toxicity associated with this kind of treatment but is insufficient to determine the individual response in each patient or to tailor the dose to each patient's real requirements. In the field of solid organ transplantation, new approaches able to reflect the individual responses produced in each patient are required to monitor immunosuppressive therapy and to improve the efficacy and safety of treatment. Thus, in the last few years, preliminary studies evaluating new specific biomarkers of the biologic effect of immunosuppressive drugs (pharmacodynamic monitoring) have been carried out. The results of these pilot studies show the potential of monitoring of these specific biomarkers to improve the safety and efficacy of immunosuppression, although the complexity of these analyses and the lack of standardized methodologies currently limit the routine application of this technique. Based on the previous studies performed to date on biomarkers, which included a small number of patients, no conclusions can be drawn as to which are the most appropriate biomarkers to prevent organ rejection or adverse events. Consequently, these biomarkers need to be validated in multicenter clinical trials. The present article reviews some biomarkers that have been proposed to evaluate currently approved immunosuppressants, such as target enzymes, cytokines, lymphocyte activation biomarkers, and cellular immune response, as well as the most promising results in this field. PMID- 19298944 TI - Natural products in medicinal chemistry. PMID- 19298945 TI - T cell memory. PMID- 19298948 TI - New JBI emphasis on translational bioinformatics. PMID- 19298946 TI - Biochemical signaling pathways for memory T cell recall. AB - Memory T cells exhibit low activation thresholds and rapid effector responses following antigen stimulation, contrasting naive T cells with high activation thresholds and no effector responses. Signaling mechanisms for the distinct properties of naive and memory T cells remain poorly understood. Here, I will discuss new results on signal transduction in naive and memory T cells that suggest proximal control of activation threshold and a distinct biochemical pathway to rapid recall. The signaling and transcriptional pathways controlling immediate effector function in memory T cells closely resemble pathways for rapid effector cytokine production in innate immune cells, suggesting memory T cells use innate pathways for efficacious responses. PMID- 19298949 TI - Perceptual simulation in conceptual combination: evidence from property generation. AB - In three experiments, participants received nouns or noun phrases for objects and verbally generated their properties ("feature listing"). Several sources of evidence indicated that participants constructed perceptual simulations to generate properties for the noun phrases during conceptual combination. First, the production of object properties for noun phrases depended on occlusion, with unoccluded properties being generated more often than occluded properties. Because a perceptual variable affected conceptual combination, perceptual simulations appeared central to combining the concepts for modifiers and head nouns. Second, neutral participants produced the same distributions of properties as participants instructed to describe images, suggesting that the conceptual representations used by neutral participants were similar to the mental images used by imagery participants. Furthermore, the property distributions for neutral and imagery participants differed from those for participants instructed to produce word associations. Third, participants produced large amounts of information about background situations associated with the object cues, suggesting that the simulations used to generate properties were situated. The experiments ruled out alternative explanations that simulation effects occur only for familiar noun phrases associated with perceptual memories and that rules associated with modifiers produce occlusion effects. A process model of the property generation task grounded in simulation mechanisms is presented. The possibility of integrating the simulation account of conceptual combination with traditional accounts and well-established findings is explored. PMID- 19298950 TI - Corneal wavefront errors 24 months after deep lamellar endothelial keratoplasty and penetrating keratoplasty. AB - PURPOSE: To evaluate high-order aberrations (HOA) induced by the anterior corneal surface after deep lamellar endothelial keratoplasty (DLEK) and penetrating keratoplasty (PK). DESIGN: Prospective, randomized clinical trial. METHODS: Twenty-eight eyes of 25 patients with corneal edema resulting from Fuchs dystrophy underwent DLEK with a 9- to 10-mm incision (n = 13) or PK with double running sutures (n = 15) at the Cornea Service, Mayo Clinic Department of Ophthalmology, Rochester, Minnesota. The main outcome measures were HOA from the anterior corneal surface calculated from corneal topography and decomposed into Zernike polynomials to the sixth order, high- and low-contrast visual acuity (VA), and contrast sensitivity. Variables after surgery were compared with those before surgery and between treatments by using generalized estimating equation models with Bonferroni adjustment. RESULTS: Total HOA through 24 months (0.48 +/- 0.15 microm) after DLEK was similar to total aberration before surgery (0.44 +/- 0.23 microm; P = .10). After PK, total HOA remained elevated through 24 months (1.68 +/- 0.58 microm) compared with that before surgery (0.49 +/- 0.27 microm; P < .005) and compared with that after DLEK (P < .006). At 24 months after PK, corneas with sutures removed had greater total HOAs than corneas with sutures intact (1.90 +/- 0.52 microm vs 1.18 +/- 0.33 microm; P = .001). High- and low contrast VA and contrast sensitivity at 24 months after PK did not correlate with any HOA. CONCLUSIONS: HOAs from the anterior corneal surface were higher after PK compared with after DLEK but did not correlate with visual function after PK. PMID- 19298951 TI - [Pathology of adrenocortical tumors: review and recent data]. AB - Most adrenocortical tumors are benign; adrenocortical carcinomas are rare but their prognosis is poor and their therapeutic is sparse. In most adrenocortical tumors, the morphological approach brings sufficient elements to establish the differential diagnosis between a benign and a malignant tumor but in few cases, it is insufficient. Moreover, morphology is limited for predicting prognosis of adrenocortical carcinomas. These observations led to development of other approaches, in particular genetic approaches. These genetics findings already have repercussions for the patients in the development of molecular markers for diagnosis and prognosis and in the future they could help in the development of new morphological approaches, in particular immunohistochemical approaches. PMID- 19298952 TI - Chitosan derivatives alter release profiles of model compounds from calcium phosphate implants. AB - The aim of the current study was to evaluate the impact of chitosan derivatives, namely N-octyl-chitosan and N-octyl-O-sulfate chitosan, incorporated in calcium phosphate implants to the release profiles of model drugs. The rate and extent of calcein (on M.W. 650 Da) ED, and FITC-dextran (M.W. 40 kDa) on in vitro release were monitored by fluorescence spectroscopy. Results show that calcein release is affected by the type of chitosan derivative used. A higher percentage of model drug was released when the hydrophilic polymer N-octyl-sulfated chitosan was present in the tablets compared with the tablets containing the hydrophobic polymer N-octyl-chitosan. The release profiles of calcein or FD from tablets containing N-octyl-O-sulfate revealed a complete release for FD after 120 h compared with calcein where 20% of the drug was released over the same time period. These results suggest that the difference in the release profiles observed from the implants is dependent on the molecular weight of the model drugs. These data indicate the potential of chitosan derivatives in controlling the release profile of active compounds from calcium phosphate implants. PMID- 19298953 TI - Study on the antiendotoxin action of Pulsatillae Decoction using an Affymetrix rat genome array. AB - A high-throughput and efficient Affymetrix rat genome array was used to investigate the pharmacological mechanism of the traditional Chinese medicine, Pulsatillae Decoction (PD), used for the treatment of diseases induced by lipopolysaccharide (LPS). Rat intestinal microvascular endothelial cells (RIMECs) were challenged with 1mug/ml LPS for 3h, and then treated with PD at a concentration of 1mg/ml for 24h. Total RNA from each treatment group was extracted from cultured RIMECs for detection by the Affymetrix Rat Genome 230 2.0 Array. The results showed that 36 genes were upregulated and 33 genes were downregulated in the LPS group vs. the blank control group; 566 genes were upregulated and 12 genes were downregulated in the PD-treated group vs. the LPS group; and 93 genes were upregulated and 29 genes were downregulated in the PD treated group vs. the blank control group. The analysis of these data suggested that PD specifically and effectively reduce damage induced by LPS, and improved physiological and biochemical responses to counteract the effects of LPS. PMID- 19298954 TI - [Practical issues in the management of third and fourth degree tears to minimise the incidence of faecal incontinence]. AB - Faecal incontinence is underestimated in most epidemiological studies, probably because women may feel unable to discuss the symptoms they experience and avoid seeking medical advice. The most common cause of faecal incontinence in healthy women is an injury during a vaginal delivery. In this article we review the classification and terminology, as well as the risk factors, for third and fourth degree perineal tears. We also comment on the different suture techniques, the follow-up of women who sustain third and fourth degree tears and the advice given to those women regarding future pregnancies and mode of delivery. We highlight the importance of the endoanal ultrasound on the diagnosis of occult anal sphincter injury. PMID- 19298955 TI - [Rectourethral fistula due to a locally advanced rectal tumour]. PMID- 19298956 TI - [General surgeons and varicose vein surgery]. AB - Chronic venous insufficiency is a highly prevalent condition, with significant health and economic repercussions. Although important therapeutic developments have been introduced in recent years, the majority are dealt with by general surgeons in national health hospitals. These surgeons do not have the required and continuous training, and continue to perform classic surgery techniques. Also, their presence at scientific, organisational meetings and training is almost nil. We present an update on developments in phlebology, and tapping into the preliminary results of a national survey, we reflect on the current status of phlebology and beyond for those general surgeons who should have a role in this field. PMID- 19298957 TI - [Primary abdominal wall actinomycosis]. PMID- 19298958 TI - [Diverticulitis due to a giant Meckel diverticulum containing a fecalith]. PMID- 19298959 TI - [Treatment of complex enterocutaneous fistulas using the open vacuum-pack technique as a better therapeutic option]. PMID- 19298960 TI - [Importance of physical examination and imaging techniques in the diagnosis of anorectal fistulae]. AB - AIM: The study was designed to determine the role of clinical examination and imaging techniques in the diagnosis of anorectal fistula. MATERIAL AND METHODS: We performed an observational study with prospective recruiting using the data of 120 patients, by means of clinical evaluation by an experienced coloproctologist surgeon (EE), a surgeon without special training in coloproctology (CE), and examination under anaesthesia (SE), endoanal ultrasound (EAU) and magnetic resonance (MR), using the surgical findings as a reference. RESULTS: SE was significantly better than EE or CE for detecting an internal opening (IO), primary track and abscess cavities (AC). EAU was significantly more sensitive and accurate than the EE in identifying an IO, and AC, but not compared to the SE. MR was more sensitive than the EE in the identification of the IO, transphincter and suprasphincter tracks and AC with no significant differences compared to EAU, and more sensitive than the SE to detect AC. CONCLUSIONS: Examination under anaesthesia still has a place in the evaluation of anorectal fistula. Imaging methods are an occasional complement to a clinical evaluation that can help the less experienced to decide the appropriate treatment, particularly when a complex fistula is suspected. PMID- 19298961 TI - Activating event knowledge. AB - An increasing number of results in sentence and discourse processing demonstrate that comprehension relies on rich pragmatic knowledge about real-world events, and that incoming words incrementally activate such knowledge. If so, then even outside of any larger context, nouns should activate knowledge of the generalized events that they denote or typically play a role in. We used short stimulus onset asynchrony priming to demonstrate that (1) event nouns prime people (sale shopper) and objects (trip-luggage) commonly found at those events; (2) location nouns prime people/animals (hospital-doctor) and objects (barn-hay) commonly found at those locations; and (3) instrument nouns prime things on which those instruments are commonly used (key-door), but not the types of people who tend to use them (hose-gardener). The priming effects are not due to normative word association. On our account, facilitation results from event knowledge relating primes and targets. This has much in common with computational models like LSA or BEAGLE in which one word primes another if they frequently occur in similar contexts. LSA predicts priming for all six experiments, whereas BEAGLE correctly predicted that priming should not occur for the instrument-people relation but should occur for the other five. We conclude that event-based relations are encoded in semantic memory and computed as part of word meaning, and have a strong influence on language comprehension. PMID- 19298962 TI - Galileo's vision. PMID- 19298963 TI - Divided hemispheres. PMID- 19298964 TI - Adrenal androgen dehydroepiandrosterone sulfate inhibits vascular remodeling following arterial injury. AB - Recent epidemiologic studies have suggested that serum dehydroepiandrosterone sulfate (DHEAS) levels have a significant inverse correlation with the incidence of cardiovascular diseases. However, direct evidence for the association with DHEAS and vascular disorders has not yet been explored. DHEAS significantly reduced neointima formation 28 days after surgery without altering other serum metabolite levels in a rabbit carotid balloon injury model. Immunohistochemical analyses revealed the reduction of proliferating cell nuclear antigen (PCNA) index and increase of TdT-mediated dUTP-biotin Nick End Labeling (TUNEL) index, expressing differentiated vascular smooth muscle cell (VSMC) markers in the media 7 days after surgery. In vitro, DHEAS exhibited inhibitory effects on VSMC proliferation and migration activities, inducing G1 cell cycle arrest with upregulation of one of the cyclin dependent kinase (CDK) inhibitors p16(INK4a) and apoptosis with activating peroxisome proliferator-activated receptor (PPAR) alpha in VSMCs. DHEAS inhibits vascular remodeling reducing neointima formation after vascular injury via its effects on VSMC phenotypic modulation, functions and apoptosis upregulating p16(INK4a)/activating PPARalpha. DHEAS may play a pathophysiological role for vascular remodeling in cardiovascular disease. PMID- 19298965 TI - One step carbon nanotubes-based solid-phase extraction for the gas chromatographic-mass spectrometric multiclass pesticide control in virgin olive oils. AB - This article presents a novel application of carbon nanotubes for the determination of pesticides (chlortoluron, diuron, atrazine, simazine, terbuthylazin-desethyl, dimetoathe, malathion and parathion) in virgin olive oil samples. For this purpose, two carbon nanotubes, multi-walled and carboxylated single-walled, were evaluated, the later being the most appropriate for the aim of the work. The sorbent (30 mg) was packed in 3-mL commercial cartridge and the virgin olive oil samples diluted (20%, v/v) in hexane were passed through it. After a washing step with 3 mL of hexane to remove the sample matrix, the pesticides were eluted with 500 microL of ethyl acetate. In order to achieve lower detection limits, the eluent was evaporated under a nitrogen stream and the residue reconstituted in 50 microL of the same solvent. Aliquots of 2 microL of the extract were directly injected into the GC-MS system for analysis. The low limits of detection achieved, between 1.5 and 3.0 microg L(-1), permit the application of the method to control the presence of these pollutants in very restrictive samples such as the ecological virgin olive oil. In addition to the sensitivity enhancement, the solid-phase extraction procedure is rather simple as it involves a single preconcentration-elution step, which allows sample processing in less than 8 min. Moreover, the cartridge can be reused at least 100 times without losing performance. The method was applied to the determination of the pesticides in two monovarietal and one ecologic commercial extra virgin olive oil samples. Two pesticides were detected in each of the monovarietal virgin olive oils while the ecological sample resulted to be a pesticide-free one. PMID- 19298966 TI - Improvement of chromatographic performances of in-situ synthesized hybrid C8 silica monoliths by reduction of structural radial heterogeneities. AB - Several modifications of a previously described protocol are proposed to improve the performances of in-situ synthesized C(8) hybrid silica monoliths. Our attention was focused on reducing the sources of radial heterogeneity that may be responsible for the poor efficiencies observed in the hydrodynamic elution mode. It was demonstrated that a decrease in the temperature of the capillary during the filling step equally to that of the polymerization mixture (0 degrees C), associated with a decrease of the gelation temperature to 20 degrees C along with a new pre-treatment of the capillary's internal walls [with a mixture of tetraethoxysilane (TEOS)/EtOH (1/3, v/v)] allows (i) increasing the radial homogeneity of the monolith, thus further enhancing the performances in the nano liquid chromatography (nano-LC) mode, (ii) improving the capillary to capillary reproducibility in terms of permeability and efficiencies. In fact, the average minimum plate height H(min) was lowered from 24 to 14 microm and the capillary-to capillary reproducibility of the synthesis was widely improved by factors two and three of reduction on the calculated standard deviation, respectively for both the efficiency in the nano-LC mode and the permeability. At last, the improved radial homogeneity and anchoring of the synthesized monoliths allowed increasing the inner diameter of the capillary (up to 150 microm) without any significant loss in efficiency. Finally, long term stability of the as-obtained monolithic stationary phases in terms of retention and efficiency was studied. In addition, the evaluation of their chromatographic behaviour was also achieved with the Tanaka test and the results were compared to those already published for commercial monoliths (Chromolith) as well as for particulate stationary phases. PMID- 19298967 TI - Single and two-component cation-exchange adsorption of the two pure major whey proteins. AB - Adsorption of pure alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) to the cation exchanger SP Sepharose FF was studied at pH 3.7 with the purpose of developing a process for isolating them from whey. Measurement of Langmuir parameters describing adsorption equilibrium in batch experiments and protein breakthrough time values in 1-ml packed-beds at a linear velocity of 158 cm/h and initial concentrations of 3 mg/ml for BLG and 1.5 mg/ml for ALA suggested the feasibility of using this adsorbent to separate the two proteins when present in a mixture. Subsequent experiments with 5-ml columns at the above concentrations and a linear velocity of 30 cm/h confirmed this and showed evidence of competitive adsorption as ALA displaced and eluted all BLG from the column in a pure form, and the remaining ALA could be eluted thereafter at high purity and with 91% recovery. PMID- 19298968 TI - Rapid screening for anthocyanins and anthocyanin dimers in crude grape extracts by high performance liquid chromatography coupled with diode array detection and tandem mass spectrometry. AB - A rapid and efficient method using high performance liquid chromatography coupled with diode array detection and tandem mass spectrometry (HPLC-DAD-MS/MS) for fast screening large numbers of anthocyanins and anthocyanin dimers in different grape skin extracts, without further sample clean-up procedures, was developed. A good separation of most detected anthocyanins was achieved in a run time of 15 min. Identification of anthocyanin pigments required a combination of several information: UV-vis spectra, MS and MS/MS spectra, and elution pattern. Many compounds have been here detected for the first time and their structures tentatively elucidated. PMID- 19298969 TI - Characterization of benzothiazoles, benzotriazoles and benzosulfonamides in aqueous matrixes by solid-phase extraction followed by comprehensive two dimensional gas chromatography coupled to time-of-flight mass spectrometry. AB - Benzothiazoles, benzotriazoles and benzosulfonamides are high-production-volume chemicals found in various environmental aqueous samples that should be considered as emerging pollutants. This study examines the suitability of comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC x GC-TOF-MS) for the characterization of benzothiazoles, benzotriazoles and benzosulfonamides in aqueous matrices. Solid-phase extraction was optimized in order to ensure the proper preconcentration of these contaminants prior to their analysis. Column selection, in both the first and second dimensions, was optimized to ensure a good chromatographic separation of the target analytes and of the potential interfering compounds extracted from the matrix. Several column phases were tested. The combined power of two-dimensional separation was fully illustrated by identifying minor compounds and avoiding the overestimations usually made by one-dimensional systems. Finally, the suitability of GC x GC-TOF-MS to quantify the aforementioned emerging pollutants was proven by determining those pollutants in several real samples such as river water, effluent from a wastewater treatment plant, and raw sewage. PMID- 19298970 TI - Cerebral segmental polyarteritis of unknown aetiology in sheep. AB - Vasculitis affecting the meningeal elastic arteries was identified in six sheep of various breeds during routine diagnostic investigation. The lesions consisted of multifocal intimal proliferation, thickening of the tunica media, multifocal infiltration of the media by inflammatory cells and more extensive perivascular accumulation of lymphocytes and plasma cells within the tunica adventitia. Fibrinoid necrosis affected an intergyral artery in one sheep. Immunohistochemistry failed to demonstrate pestivirus antigen associated with the lesions and no alternative aetiology was defined. Possible causes of meningeal segmental polyarteritis in sheep are discussed. PMID- 19298971 TI - Speech and language disorders secondary to diffuse subcortical vascular lesions: Neurolinguistic and acoustic analysis. A case report. AB - BACKGROUND AND PURPOSE: Subcortical white matter (WM) plays an important role in speech production and language processing. Most frequently, cerebral WM lesions are secondary to small vessel disease in patients with vascular risk factors. We report the case of a 53-year-old man with history of hypertension and ischemic subcortical lesions, who presented with speech difficulties and mild cognitive impairment. METHODS: Language and cognitive assessment included Boston Diagnostic Aphasia Examination, Boston Naming Test, Rey Auditory-Verbal Learning Test, Rey Osterrieth Complex Figure Test, Trail Making Test A and B, Wisconsin Card Sorting Test, Scale for Evaluation of Perceptive Characteristics of Voice and Speech, and Multidimensional Evaluation of Speech and Voice. RESULTS: Brain MRI showed ischemic WM lesions and lacunar infarcts in the brainstem and right cerebellum. Cognitive testing revealed mild cognitive impairment, predominantly affecting attention and executive functions. Speech and language analysis demonstrated dysarthria, dysphonia with hypophonia, and imprecise articulation, as well as short rushes of speech, palilalia and mild subcortical dysphasia. CONCLUSIONS: Neurolinguistic and acoustic analysis in patients with ischemic WM lesions can provide additional information in the understanding of language and speech disturbances, and can assist in patient management. PMID- 19298974 TI - Functional and oncological outcomes of partial nephrectomy of solitary kidneys. AB - PURPOSE: We examined outcomes after partial nephrectomy in patients with tumors in a solitary kidney to determine the extent to which patient, surgery and tumor specific variables influenced the glomerular filtration rate and local recurrence postoperatively. MATERIALS AND METHODS: Demographics, renal function, comorbidities, renal cell carcinoma history, and operative and pathological data were recorded. The effect on changes in early and late postoperative glomerular filtration rate and local recurrence were analyzed. RESULTS: In 84 patients undergoing a total of 89 partial nephrectomies the mean immediate postoperative decrease in the glomerular filtration rate in those with no ischemia, warm ischemia (mean 12 minutes) and cold ischemia (mean 33 minutes) was 29%, 37% and 45%, respectively (p <0.01). Late glomerular filtration rate decreases were 12%, 6% and 16%, respectively (p = 0.17). Cold ischemia and multiple vascular risk factors were associated with immediate glomerular filtration rate decreases (p = 0.008 and 0.04, respectively). Local recurrence, which developed in 13 patients (18%), was associated with positive margins and T stage (p = 0.01 and 0.02, respectively). End stage renal disease developed in 3 patients (4%) and an additional 5 (6%) required nephrectomy for local recurrence. CONCLUSIONS: Partial nephrectomy generally results in a small decrease in the glomerular filtration rate, and limited warm and cold ischemia does not appear to adversely affect long term renal function. Positive margins and T stage greater than 2 are the most important predictors of local recurrence in a solitary kidney. They pose a significant risk to end stage renal disease-free survival due to the need for completion nephrectomy in many of these patients. Partial nephrectomy should be considered the standard of care in all patients with tumor in the solitary kidney. PMID- 19298975 TI - Tetrabromobisphenol A: investigating the worst-case scenario. PMID- 19298976 TI - [Hospital management of hyperglycemia]. PMID- 19298977 TI - [Predictive value of MRI for detecting and characterizing vascular compression in cranial nerve hyperactivity syndromes (trigeminal and facial nerves)]. AB - MRI detects vascular compression of the cranial nerve in the majority of the cases. High-resolution 3D-T1 and 3D-T2 MRI gives detailed images, particularly the 3D-T2 MRI sequences, with good contrast between cerebrospinal fluid and vascular and nerve structures. TOF-AMR (native sequence and vertebrobasilar reconstruction) shows the vascular structures in hypersignal and therefore differentiates the vessels from the cranial nerves. The 3D-T1 sequence with gadolinium reinforces the signal of the venous structures. Thus, preoperative MRI makes it possible to predict the existence of a vascular compression. The correlation study between imaging data and intraoperaitive anatomical findings showed a sensitivity of MRI of 97% and a specificity of 100%. In addition, it can specify the type and the degree of the compression. This information may help in selecting the most appropriate surgical method. PMID- 19298978 TI - [Functional anatomy of the facial nerve]. AB - Embryologic individualization of the facial nerve primordium occurs early and emphasizes trigeminofacial connections and variations in the transitional zone (TZ). In the brainstem, the specific nuclei of the facial nerve are located within five columns corresponding to the main functions. Three-quarters of the fibers are of the special visceral efferent group (SVE). Central connections of the facial motonucleus are numerous, particularly with the trigeminal complex but also the cochlear nerves. Only endoneurium surrounds facial nerve fibers in the cerebellopontine angle; central nervous system tissue offers a small extracellular space, lacking collagen and funicular plexus, without epiperineurium: nerve fibers are therefore more susceptible to injury. The segment of a nerve that reveals both CNS and PNS components is referred to as the transition zone. It should be regarded as a "locus minor resistae" because of poor vascularization and CNS myelin deficiency. The acousticofacial reflex is often absent in patients with hemifacial spasm. Early (R1) and late (R2) responses of the trigeminofacial reflex (blink reflex) in patients with hemifacial spasm are of major interest in understanding both peripheral and central mechanisms. PMID- 19298979 TI - [Balloon compression for the treatment of trigeminal neuralgia]. AB - The authors present a retrospective study of 121 patients treated with balloon compression of the rootlets behind the Gasser ganglion from 1995 to 2007. The inclusion criteria were drug-resistant idiopathic trigeminal neuralgia. The authors described the surgical technique and compared their results with results from the literature. The following parameters were compared: technical success, pain relief and recurrence, and complications. Balloon compression is considered in the literature to be a safer procedure than other percutaneous surgeries, especially for postoperative sensitive disorders. The best indications seem to be trigeminal neuralgia in older patients or pain due to multiple sclerosis and neuralgia involving the V1 territory. These conclusions should be confirmed by larger series, a longer follow-up (> 5 years) and statistically better analysis. PMID- 19298980 TI - [Electrophysiological monitoring of cranial motor nerves (V, VII, IX, X, XI, XII)]. AB - In various neurosurgical operations, there is a risk of cranial nerve lesion that can be avoided or minimized with intraoperative electrophysiological monitoring. Regarding motor function of the cranial nerves, stimulodetection techniques are used, including electrical stimulation of nerve trunks and electromyographic recording of evoked motor responses. These techniques can be used for monitoring the trigeminal nerve (Vth cranial nerve), facial nerve (VIIth), glossopharyngeal nerve (IXth), pneumogastric nerve (Xth), spinal accessory nerve (XIth), and hypoglossal nerve (XIIth), in particular during surgical removal of tumors of the cerebellopontine angle or skull base. When beginning an operation, electrical stimulation is only used to identify the nerve structures. As removal of the tumor progresses, the goal is to verify that a surgical injury to the nerve is avoided by looking for the absence of any change regarding amplitude, morphology, and latency of motor responses. Intraoperative electromyographic monitoring can also be applied during the surgical treatment of primary hemifacial spasm by microvascular decompression. An effective decompression is usually associated with the disappearance of "lateral spread" motor responses to facial nerve branch stimulation. Therefore, the intraoperative disappearance of the lateral spread responses can be considered a predictive factor of good postoperative clinical outcome, even if this assertion remains a matter of debate. PMID- 19298981 TI - [Neurosurgical treatment of vago-glossopharyngeal neuralgia]. AB - Glossopharyngeal neuralgia, more accurately called vago-glossopharyngeal neuralgia (VGPN) because of the frequent association with pain irradiation in the sensory territory of the vagus nerve, is not always recognized because its incidence is much lower than the incidence of trigeminal neuralgia (100 times more frequent). As in trigeminal neuralgia, when pain becomes resistant to anticonvulsants - its specific medical treatment - VGPN can almost always be cured by surgery. The first option is microvascular decompression, since vascular compression is the main cause of the neuralgia. Percutaneous thermorhizotomy at the foramen jugularis (pars nervosa) is only indicated as a second option, because of unavoidable sensorimotor deficits in the ninth and tenth nerves. Tractonucleotomies at the medullary level should be reserved essentially for pain of malignant origin. PMID- 19298982 TI - [Intraoperative monitoring of brainstem auditory evoked potentials during microvascular decompression of cranial nerves in cerebellopontine angle]. AB - Brainstem auditory evoked potentials (BAEP) monitoring is a useful tool to decrease the danger of hearing loss during pontocerebellar angle surgery, particularly in microvascular decompression (MVD). Critical complications arising during MVD surgery are the stretching of the VIII nerve - the main cause of hearing loss - labyrinthine artery manipulation, direct trauma with instruments, or a nearby coagulation, and at end of the surgery neocompression of the cochlear nerve by the prosthesis positioned between the conflicting vessel(s) and the VIIth-VIIIth nerve complex. All these dangers warrant the use of BEAP monitoring during the surgical team's training period. Based on delay in latency of peak V, we established warning thresholds that can provide useful feedback to the surgeon to modify the surgical strategy: the initial signal at 0.4 ms is considered the safety limit. A second signal threshold at 0.6 ms (warning signal for risk) corresponds to the group of patients without resultant hearing loss. The third threshold characterized by the delay of peak V is at 1 ms (warning signal for a potentially critical situation). BAEP monitoring provides the surgeon with information on the functional state of the auditory pathways and should help avoid or correct manoeuvres that can harm hearing function. BAEP monitoring during VIIth-VIIIth complex surgery, particularly in MVD of facial nerves for HFS is very useful during the learning period. PMID- 19298983 TI - [Functional assessment of cochlear nerve and hearing]. AB - BACKGROUND/PURPOSE: Since otoneurosurgical pathologies are being diagnosed increasingly early, the functional prognosis of surgery is gaining in importance. Hearing and cochlear nerve exploration can be carried out using three principal methods: pure-tone and speech audiometry, evoked otoacoustic emissions (EOAEs), and brainstem auditory evoked potentials (BAEPs). The aim of this paper is to present the current knowledge on auditory functional assessment. MATERIALS AND METHODS: The medical and scientific literature from the Pubmed-Medline database was reviewed. Recently published related books were also included. RESULTS/DATA SYNTHESIS: Preoperative audiological evaluation makes it possible to assess pathology consequences on hearing. Depending on auditory status, conservative surgery, including intraoperative auditory monitoring, can be planned. From this perspective, prognostic factors for hearing preservation - such as the speech intelligibility score, the presence of transient EOAEs, and the presence of the BAEPs wave III - were identified. However, several surgical teams argue for a systematic conservative procedure, particularly when EOAEs are present, whatever the patient's preoperative hearing status. Postoperative audiological evaluation provides the opportunity for monitoring: in the short term, to detect complications early, through BAEPs recorded in the nonoperated ear; in the long term, to detect recurrence of the pathology. Moreover, the consequences of surgery on hearing can be assessed and long-term follow-up of auditory function can be carried out. Finally, the comparison of pre- and postoperative audiological status provides inductive feedback on surgery and electrophysiological monitoring, giving the otoneurosurgeon and neurophysiologist the opportunity to improve the entire procedure. CONCLUSION: Auditory functional assessment is essential for optimal care of tumor or functional pathologies involving the cochlear nerve and/or the cerebellopontine angle. PMID- 19298984 TI - Antimalarial sesquiterpene lactones from Distephanus angulifolius. AB - Combined use of bioassay-guided fractionation based on in vitro antiplasmodial assay and dereplication based on HPLC-PDA-MS-SPE-NMR led to isolation of (6S,7R,8S)-14-acetoxy-8-[2-hydroxymethylacrylat]-15-helianga-1(10),4,11(13)-trien 15-al-6,12-olid and (5R,6R,7R,8S,10S)-14-acetoxy-8-[2-hydroxymethylacrylat]-elema 1,3,11(13)-trien-15-al-6,12-olid, along with vernodalol, vernodalin, and 11,13beta-dihydroxyvernodalin from extract of Distephanus angulifolius. All compounds were identified by spectroscopic methods, including 1D and 2D homo- and heteronuclear NMR experiments. The isolated compounds showed IC(50) values in the range 1.6-3.8 microM and 2.1-4.9 microM against chloroquine sensitive D10 and chloroquine resistant W2 Plasmodium falciparum strains, respectively. PMID- 19298985 TI - [Mediastinal paragangliomas: a report of 2 cases]. AB - Paragangliomas are neuroendocrine tumors derived from chromaffin cells of the extraadrenal sympathetic system; mediastinal paragangliomas are rare. We illustrate and describe the imaging findings in two cases of mediastinal paraganglioma, one nonfunctional aorticopulmonary paraganglioma and one locally aggressive functional aorticosympathetic paraganglioma first observed as a mediastinal mass on chest plain-film radiographs. PMID- 19298986 TI - [Desmoplastic small round cell tumor of the abdomen: CT findings and radiologic pathologic correlation in 3 cases]. AB - We describe the CT findings of abdominal desmoplastic small round cell tumor (DSRCT) in three patients. The most common imaging finding was the presence of single or multiple soft-tissue density intraperitoneal masses without apparent origin in an abdominal organ. In the first patient, a single peritoneal mass was located in the mesentery between the stomach and pancreas. In the second patient, an intraperitoneal pelvic mass was seen in the retrovesical space. In the third patient, a large homogeneous soft-tissue mass that nearly filled the entire peritoneal space was found. Two patients had multiple liver metastases and adenopathies at the time of diagnosis. The diagnosis was confirmed with CT-guided percutaneous biopsies in all three cases. DSRCT should be suspected in young men with multiple bulky heterogeneous soft-tissue masses. PMID- 19298987 TI - Relationship between biochemical markers and radiographic scores in the evaluation of the osteoarticular status of Warmblood stallions. AB - Establishing the osteoarticular status of the horse is often performed by means of radiological screening of the animals. Widespread blood sampling could reveal to be an interesting alternative to this procedure which is time consuming and sometimes technically difficult. The aim of this study was to investigate the relationship between the radiological status of the horses and the levels of biochemical markers of cartilage degradation and synovial inflammation. A specific radiological scoring and classification system was therefore developed and applied on 63 stallions presented for studbook admission. Additionally, groups of horses were established according to the occurrence of osteochondrosis, degenerative joint disease and distal interphalangeal joint effusion. Insulin growth factor-I, myeloperoxidases, Coll2-1 and Coll2-1NO(2) were used as blood markers. The combination of the blood parameters did not seem to correlate with the used scoring system. Coll2-1NO(2) levels however tended to increase with poorer radiological class and this could therefore potentially be a useful predictor of the osteoarticular status in the horse. Coll2-1 levels were significantly higher in the degenerative joint disease group. A high percentage of horses with distal interphalangeal joint effusion was present in this study and was associated with decreased IGF-I and increased Coll2-1 levels. PMID- 19298988 TI - Comparison of PCR methods for diagnosis of canine visceral leishmaniasis in conjunctival swab samples. AB - Four PCR assays for detection of Leishmania DNA in conjunctival swab samples were compared. All methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or seminested). Two methods (kDNA PCR-hybridization and kDNA snPCR) used primers targeted to the minicircles of kinetoplast DNA (kDNA) and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of ribosomal rRNA genes. kDNA PCR hybridization was positive for 22/23 dogs (95.6%) and for 40/46 samples (86.9%), considering the right and the left conjunctivas. kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The ITS-1 nPCR and LnPCR were both able to detect the parasites in 17/23 dogs (73.9%) and 29/46 (63%) and 30/46 (65.2%) samples, respectively. The positivities of the kDNA based methods were significantly higher; however the choice of the best method will depend on the kind of information required with the diagnosis. PMID- 19298989 TI - Infection by Eucoleus aerophilus in dogs and cats: is another extra-intestinal parasitic nematode of pets emerging in Italy? AB - The occurrence of the infection by the lungworm Eucoleus aerophilus (syn. Capillaria aerophila) in dogs and cats from Italy has been evaluated with conventional diagnostic procedures. Individual faecal samples from 569 dogs and 200 cats were undertaken to faecal flotation with sugar and zinc sulphate solution. Sixteen dogs (2.8%) and 11 cats (5.5%) scored positive for eggs of E. aerophilus when samples were processed with either of the two flotation solutions. Overall 14 of 16 dogs and eight of 11 cats infected by E. aerophilus showed respiratory symptoms and the most common were general respiratory distress, dry cough, wheezing and sneezing. These results indicate that E. aerophilus is not uncommon and that canine and feline capillariosis is of clinical importance. Given the impact that E. aerophilus infections may have upon animal health and its zoonotic potential, it is strongly advisable to routinely include this disease in the differential diagnosis of (cardio)-respiratory diseases of dogs and cats. PMID- 19298990 TI - Cross-sectional anatomy of the rabbit neck and trunk: comparison of computed tomography and cadaver anatomy. AB - Computed tomographic images of the neck, thorax and abdomen in four healthy adult rabbits were obtained with a conventional CT using a slice-thickness of 5mm. CT images were obtained with the animals positioned in sternal recumbency on a removable plastic support directly laying on the CT-table. At the end of the CT session, each rabbit was euthanized and, while carefully maintaining the same position on the plastic support, the animal was moved into a -20 degrees C freezer until completely frozen. Each cadaver was then sectioned at 10mm slices, with the first section starting at the tip of the nose, respecting the imaging protocol. The frozen sections were cleaned and then photographed on each side. Anatomic structures were identified and labeled first on each side of the frozen section and then on the corresponding CT image with the aid of the available literature. Results from our study provide an atlas of normal cross-sectional gross and CT anatomy of the rabbit neck, thorax and abdomen, useful in the interpretation of any cross-sectional imaging modality in this species. PMID- 19298991 TI - [Fetal cerebral magnetic resonance imaging (MRI). Indications, normal and pathological patterns]. AB - Fetal MRI is a specific imaging modality, always performed after a reference ultrasound examination. The decision to perform an MRI-scan must take into account the anxiety constantly generated by the need for this unusual examination during pregnancy. To date, no side-effect associated with 1.5 tesla magnets has been described. Compared to ultrasonography, fetal brain MRI provides better contrast between grey and white matter, as well as better delineation of the brainstem (pontic curvature) and the cerebellum (lobules and fissures). However, it often remains difficult to inform parents about prognosis. Thereby, it is of utmost importance to be familiar with the definite criteria associated with a poor neurological prognosis such as lack of pontic curvature or as diffuse or bilateral cortical malformations. This has to be considered within the framework of French regulations which allow pregnancy termination with no time limit. The optimal timing to perform a fetal MRI-scan depends on the context. The period between 27 and 30 weeks of gestation is a good balance between gestational age and gyration or sulcation development. The main ultrasonographic findings requiring MRI are ventriculomegalies and posterior fossa abnormalities. MRI exploration can sometimes be performed despite a normal ultrasonography in case of genetic disorders such as tuberous sclerosis and lissencephalies. In addition to its diagnostic value towards decision to terminate pregnancy, fetal MRI can be used as "in vivo autopsy", in case of expected technical difficulties or refusal of post-abortion examinations by relatives. Technical advances (real time and specific sequences like diffusion tensor and spectroscopy) and prospective clinical studies will probably improve the efficiency of this method to assess neurological prognosis. PMID- 19298992 TI - Analysis of the dose-response relationship between high-risk human papillomavirus viral load and cervical lesions. AB - The aims of this study were to explore the dose-response relationship between high-risk human papillomavirus (hrHPV) load and cervical lesions; the relationship between hrHPV viral load and the severity of cervical lesions; and the clinical application of the hybrid capture II (HC-II) system in the secondary prevention of cervical cancer. HrHPV viral load was detected by the HC-II system and cervical lesions were diagnosed from biopsied tissue. Curve estimation and Mantel trend analysis were used to explore the dose-response relationship between hrHPV viral load and cervical lesions. Spearman's rank correlation analysis and ordinal regression model were used for the analysis of hrHPV viral load and the severity of cervical lesions. Curve estimation showed good correlation between cervical lesion rates and hrHPV viral load (r=0.775, P=0.008); the rate of cervical lesions increased with hrHPV viral load (chi(trend)=8.000, P<0.001). Medium intensity rank correlation was found between hrHPV viral load grades and the severity of cervical lesions (r(s)=0.321, P<0.001); a correlation appeared between hrHPV viral load and the severity of cervical lesions (P<0.001). These results suggest a dose-response relationship between hrHPV viral load and the severity of cervical lesions. This dependence has important clinical applications and shows the potential value of the HC-II system in cervical cancer prevention. PMID- 19298993 TI - Patient retention and attrition on antiretroviral treatment at district level in rural Malawi. AB - We report on rates of patient retention and attrition in the context of scaling up antiretroviral treatment (ART) within a district hospital and its primary health centres in rural Malawi. 'Retention' was defined as being alive and on ART or transferred out, whereas 'attrition' was defined as died, lost to follow-up or stopped treatment. A total of 4074 patients were followed-up for 1803 person years: 2904 were at the hospital and 1170 at health centres. Approximately 85% of patients were retained in care, both at hospital and health centres, with a retention rate per 100 person-years of 185 and 211, respectively [adjusted hazard ratio (HR) 1.18, 95% CI 1.10-1.28, P=0.001). Attrition rates per 100 person-years were similar: 33 and 36, respectively (adjusted HR 1.17, 95% CI 0.97-1.4, P=0.1). At health centres the incidence of loss to follow-up was significantly lower than at the hospital (adjusted HR 0.24, P<0.001, risk reduction 77%), but the rate of reported deaths was higher at health centres (adjusted HR 2.2, 95% CI 1.76-2.72, P<0.001). As Malawi continues to extend the coverage (and equity) of ART, including in rural areas, attention is needed to reduce losses to follow-up at hospital level and reduce mortality at primary care level. PMID- 19298994 TI - Control of viremia and maintenance of intestinal CD4(+) memory T cells in SHIV(162P3) infected macaques after pathogenic SIV(MAC251) challenge. AB - Recent HIV vaccine failures have prompted calls for more preclinical vaccine testing in non-human primates. However, similar to HIV infection of humans, developing a vaccine that protects macaques from infection following pathogenic SIV(MAC251) challenge has proven difficult, and current vaccine candidates at best, only reduce viral loads after infection. Here we demonstrate that prior infection with a chimeric simian-human immunodeficiency virus (SHIV) containing an HIV envelope gene confers protection against intravenous infection with the heterologous, highly pathogenic SIV(MAC251) in rhesus macaques. Although definitive immune correlates of protection were not identified, preservation and/or restoration of intestinal CD4(+) memory T cells were associated with protection from challenge and control of viremia. These results suggest that protection against pathogenic lentiviral infection or disease progression is indeed possible, and may correlate with preservation of mucosal CD4(+) T cells. PMID- 19298995 TI - High titer HIV-1 V3-specific antibodies with broad reactivity but low neutralizing potency in acute infection and following vaccination. AB - Identifying the earliest neutralizing antibody specificities that are elicited following infection or vaccination by HIV-1 is an important objective of current HIV/AIDS vaccine research. We have shown previously that transplantation of HIV-1 V3 epitopes into an HIV-2 envelope (Env) scaffold provides a sensitive and specific means to detect and quantify HIV-1 V3 epitope specific neutralizing antibodies (Nabs) in human sera. Here, we employ this HIV-2/HIV-1 V3 scaffolding strategy to study the kinetics of development and breadth of V3-specific Nabs in longitudinal sera from individuals acutely infected with clade C or clade B HIV-1 and in human subjects immunized with clade B HIV-1 immunogens. HIV-2/HIV-1 chimeras containing V3 sequences matched to virus type (HIV-2 or HIV-1), subtype (clade B or C), or strain (autologous or heterologous) were used as test reagents. We found that by 3-8 weeks post infection, 12 of 14 clade C subjects had a median IC(50) V3-specific Nab titer of 1:700 against chimeric viruses containing a heterologous clade C V3. By 5 months post-infection, all 14 subjects were positive for V3-specific Nabs with median titers of 1:8000 against heterologous clade C V3 and 1:1300 against clade B V3. Two acutely infected clade B patients developed heterologous clade B V3-specific Nabs at titers of 1:300 and 1:1800 by 13 weeks of infection and 1:5000 and 1:11000 by 7 months of infection. Titers were not different against chimeras containing autologous clade B V3 sequences. Each of 10 uninfected normal human volunteers who were immunized with clade B HIV-1 Env immunogens, but none of five sham immunized control subjects, developed V3-specific Nabs titers as high as 1:3000 (median 1:1300; range 1:700 1:3000). None of the HIV-1 infected or vaccinated subjects had antibodies that neutralized primary HIV-1 virus strains. These results indicate that high-titer, broadly reactive V3-specific antibodies are among the first to be elicited during acute and early HIV-1 infection and following vaccination but these antibodies lack neutralizing potency against primary HIV-1 viruses, which effectively shield V3 from antibody binding to the functional Env trimer. PMID- 19298996 TI - Removal of phosphorus from solution using biogenic iron oxides. AB - Phosphorus removal by biogenic iron oxides was investigated, providing an initial characterization of a potentially regenerable iron-rich sorbent. The biogenic iron oxides were collected from a wetland ecosystem and were dominated by the sheaths of Leptothrix ochracea. Sorption kinetics followed a pseudo-1st order model (R(2)=0.998) with a rate constant of 0.154+/-0.013h(-1). The Langmuir isotherm adequately described sorption for all samples (R(2)=0.923-0.981); the Freundlich model was a better fit for only one of four samples. Maximum phosphorus sorption estimated using the Langmuir parameter ranged from 46.9+/-2.9 to 165.0+/-21.2mgP/gFe and was similar to other iron-rich substrates. Maximum sorption normalized to total solids ranged from 10.8+/-0.7 to 39.9+/-3.2mgP/g, which represented the highest published values for iron-rich substrates. The high sorption capacity with respect to both iron and solids warrants further evaluation of biogenic iron oxides as a substrate for phosphorus removal. PMID- 19298997 TI - Sources and prevalence of pentachlorobenzene in the environment. AB - There are no longer any large scale uses of pentachlorobenzene (PeCB). Current emissions of PeCB to the environment are estimated to be about 121000kgy(-1), based on published information. The largest sources appear to be combustion of solid wastes, 32000kgy(-1), biomass burning, 45000kgy(-1) with degradation of an agricultural fungicide, quintozene, contributing 26000kgy(-1) and industrial releases less important. PeCB has been measured in many environmental media over the past 35 years. Low but detectable concentrations of PeCB have been reported in the atmosphere, sediments and biota in remote areas of the world. Calculations using a global distribution model are consistent with the estimate of approximately 100000kgy(-1) global PeCB emissions. Concentrations of PeCB in the environment have declined with a 90% decrease of PeCB concentrations in herring gull eggs from Lake Superior, Canada since 1979. PMID- 19298998 TI - Implications of cysteine metabolism in the heavy metal response in Trichoderma harzianum and in three Fusarium species. AB - We studied the ability of four different fungal species, Trichoderma harzianum, Fusarium antophyllum, Fusarium compactum and Fusarium phyllophilum, to grow in the presence of heavy metals, and monitored their cysteine and glutathione content and the activity of O-acetylserine(thiol)lyase (OASTL), which is involved in cysteine biosynthesis. Zn and Pb did not affect fungal growth or sporulation at the concentrations used, whereas Cd and Hg did. In most cases, cysteine and glutathione content was higher when fungi were grown in the presence of toxic metals. As T. harzianum and F. phyllophilum presented the best growth rate on Cd and Hg, they were selected to further analyse the accumulation of toxic metals. Both species accumulated the four metals in large amounts. Using degenerate oligonucleotides based on fungal OASTL genes present in databases, partial OASTL DNA sequences were cloned by PCR from T. harzianum and F. phyllophilum, and the regulation of the OASTL genes under metal stress was analyzed. Cd produced the strongest increase in OASTL activity and mRNA levels in both fungi. The results suggest a possible correlation between cysteine metabolism and the heavy metal response in fungi. PMID- 19298999 TI - Metabolites from the biodegradation of triphenylmethane dyes by Trametes versicolor or laccase. AB - The feasibility of degrading triphenylmethane dyes by Trametes versicolor and laccase has been investigated for the following dyes: Acid Fuchsin, Brilliant Green 1, Basic Fuchsin, Methyl Green or Acid Green 16. The toxicity level of triphenylmethane dyes is linked to their basic character, but significant detoxification is obtained when there is biodegradation. Identification of enzymatic degradation products by (1)H NMR made it possible to propose a general rule for the laccase attack on triphenylmethane compounds. The enzyme completely degrades the molecular part of the canonical resonance substructures of dyes, because no N-substituted, mono-N and di-N,N substituted p-amine aromatic residues seem to be wholly degraded. No enzymatic degradation is observed in the cases of either the non-substituted or trisubstituted-N,N,Np-amine aromatic residues. On the other hand, for all the dyes tested, no aromatic residues are detected after fungal treatment; this means that T. versicolor is more capable of performing further degradation than is laccase. The results of this study demonstrated that compounds with a triphenylmethane structure can be degraded by T. versicolor even if they are highly toxic. The enzyme laccase plays an important role in the attack on the structure and a general rule for predicting which products would be obtained after the enzymatic treatment is suggested. PMID- 19299000 TI - Human papillomavirus profile of women in Belize City, Belize: correlation with cervical cytopathologic findings. AB - Cervical carcinoma is the most common cancer among Belizean women; however, data regarding the frequency of human papillomavirus (HPV) genotypes and their association with cervical cancer are nonexistent. We therefore included HPV genotyping as part of a week-long cervical cancer screening campaign conducted in Belize City in 2007. Conventional Papanicolaou smears with Hybrid Capture (HC) 2 HPV testing were performed on 463 women. All HC2-positive samples were genotyped using a developmental GP5+/GP6+ polymerase chain reaction-coupled Luminex assay for 2 low-risk and 18 high-risk HPV types. The prevalence of high-risk HPV was 15.6% in the total population, 10.1% in those with normal cytologic findings, and 93.3% in women with a high-grade squamous intraepithelial lesion. Of patients with HPV infections, 35% had multiple types (5.4% of the total group). Of all women and of women with normal cytologic findings, 5.2% and 2.8%, respectively, had HPV16 or 18. For all women, HPV16, 18, 56, and 52 were present in decreasing order of frequency. HPV11 was present in only one patient, and none had HPV6. HPV16 was found in 47% of high-grade squamous epithelial lesions; however, no case of HSIL had HPV18 or 45. HPV35 and HPV58 were the next most common types in high-grade squamous intraepithelial lesion, each occurring in 20% of cases of high-grade squamous intraepithelial lesion, followed by HPV31 in 13.3%. Although women younger than 25 years old were underrepresented, these data suggest that the HPV profile of this cohort of Belizean women differs somewhat from that in the region. In addition, these data are of importance with regard to the development of HPV vaccines that will be used in less developed countries, where care should be taken not to implement vaccination at the cost of basic screening and diagnostic services. PMID- 19299001 TI - Measuring in-stream retention of copper by means of constant-rate additions. AB - Human practices entail inputs of nutrients and toxicants such as heavy metals to the fluvial ecosystems. While nutrient dynamics in fluvial ecosystems have been widely studied for over three decades, dynamics of toxicants still remain unclear. In this investigation, the nutrient spiraling concept and associated methodologies to quantify nutrient retention in streams were applied to study copper (Cu) dynamics in streams. The present study aimed to quantify total dissolved Cu retention using a simplified system of indoor channels colonized with fluvial biofilms. Cu retention was studied at sub-toxic concentrations to avoid negative/lethal effects on biota. In addition, Cu retention was compared with retention estimates of a macronutrient, phosphate (PO(4)(3-)), which has been widely studied within the context of the nutrient spiraling concept. The methodology used allowed a successful quantification of Cu and PO(4)(3-) retention. The results showed higher retention efficiency for PO(4)(3-) than for Cu. The biofilm played a key role in retaining both solutes. Although retention efficiency for both solutes was higher in the experiments with colonized substrata compared to uncolonized substrata, we found a positive relationship between uptake rate and chlorophyll-a only for PO(4)(3-). Finally, retention efficiency for both solutes was influenced by water discharge, showing lower retention efficiencies under higher flow conditions. These results suggest that the fate and toxic effects of copper on stream biota may be strongly influenced by the prevailing environmental conditions. Our results indicate that the experimental approach considered can provide new insights into the investigation of retention of toxic compounds in fluvial systems and their controlling mechanisms. PMID- 19299002 TI - Increased phosphatidylserine exposure on platelets from hospitalized patients with acute medical illnesses. PMID- 19299003 TI - Association of V617F Jak2 mutation with the risk of thrombosis among patients with essential thrombocythaemia or idiopathic myelofibrosis: a systematic review. AB - INTRODUCTION: Many studies evaluated the association of V617F Jak-2 with the risk of thrombosis in patients with essential thrombocythaemia, but the results of these studies were inconsistent. Few studies evaluated the association of V617F Jak-2 mutation with the risk of thrombosis in patients with idiopathic myelofibrosis. Therefore, we performed a systematic review of the studies that assessed the risk of thrombosis associated with V617F Jak-2 in patients with ET or IM. MATERIALS AND METHODS: We searched MEDLINE and EMBASE databases and reference lists of retrieved articles. Odds ratios (ORs) and 95% confidence intervals (CI) were calculated for each trial and pooled. RESULTS: We included 21 studies involving patients with essential thrombocythaemia and 6 studies patients with idiopathic myelofibrosis. In essential thrombocythaemia patients, V617F Jak 2 was associated with a significant increased risk of thrombosis (OR 1.92, 95% CI 1.45-2.53), both of venous (OR 2.49, 95% CI 1.71-3.61) and arterial (OR 1.77, 95% CI 1.29-2.43) vessels. In idiopathic myelofibrosis patients, the risk of thrombosis associated with V617F Jak-2 tended to be increased (OR 1.76, 95% CI 0.91-3.41). CONCLUSIONS: Our systematic review suggests that V617F Jak-2 increases the risk of thrombosis in essential thrombocythaemia patients by about two fold while its role in idiopathic myelofibrosis patients is uncertain. PMID- 19299004 TI - Protective effects of iodixanol during bovine sperm cryopreservation. AB - The aim of cryopreservation is to maintain cellular integrity, thereby enabling resumption of proper biological functioning after thawing. Here we propose OptiPrep (60% iodixanol in water) as a protectant during sperm cryopreservation using pooled bull semen as the model. We evaluated OptiPrep concentration effect and its relation to cryopreservation by comparing frozen-thawed and chilled samples. Semen, extended in Andromed with 0 (control), 1.25%, 2.5%, and 5% OptiPrep, was compared after either chilling or freezing in large volume by directional freezing. Sample evaluation included sperm motility upon thawing and after 3h incubation at 37 degrees C for frozen-thawed samples and after 3h and 6h of chilling for chilled samples; viability, acrosomal integrity, and hypoosmotic swelling were also tested for frozen-thawed and chilled samples. Chilled samples with 5% OptiPrep showed inferior viability (P=0.047) and 3h motility (P=0.017) relative to that for chilled samples with 2.5% OptiPrep and inferior viability (P=0.042), acrosomal integrity (P=0.045), and 0h motility (P=0.024) relative to that for chilled samples with 1.25% OptiPrep. The 1.25%, 2.5%, and control samples did not differ. In frozen-thawed samples, 2.5% OptiPrep was superior to all other concentrations for 3h motility (control, P=0.007; 5% OptiPrep, P=0.005; 1.25% OptiPrep, P=0.004) and to 1.25% OptiPrep for acrosomal integrity (P=0.001). In a search for a protection mechanism, we measured glass transition temperature (T(g)) of Andromed and of Andromed with 1.25%, 2.5%, and 5% OptiPrep. Andromed ( 58.78 degrees C) and 1.25% OptiPrep (-58.75 degrees C) groups had lower mean T(g) than that of the 2.5% (-57.67 degrees C) and the 5% (-57.10 degrees C) groups. Directional cryomicroscopy revealed that the presence of iodixanol alters ice crystal formation into an intricate net of dendrites. Thus, iodixanol appears to possess cryoprotective properties by helping spermatozoa maintain motility and membrane integrity, possibly through altering ice crystals formation into a more hospitable environment and increasing the glass transition temperature. PMID- 19299005 TI - Group effects of instrumentality and name relation on action naming in bilingual anomic aphasia. AB - Verb production in sentences was investigated in two groups of late bilingual Greek-English speakers: individuals with anomic aphasia and a control group. Verb retrieval in sentences was significantly impaired in both languages for the individuals with anomic aphasia. Additional results revealed no effect of instrumentality on action naming in sentences in either language. However, there was a negative effect of verb-noun name relation on instrumental verb production in English only. Results confirm intact verb lemma retrieval for this group of bilingual individuals with anomic aphasia, but a breakdown at the level of accessing the phonological or lexical form. PMID- 19299007 TI - The in vitro corneal biocompatibility of hydroxyapatite-coated carbon mesh. AB - The purpose of this study was to consider the use of a hydroxyapatite (HA) coated porous carbon matrix as a synthetic dental laminate substitute in osteo-odonto keratoprosthetic (OOKP) design. 3 types of carbon meshes were coated with HA by sonoelectrochemical deposition. The materials were characterised by scanning electron microscopy (SEM) and HA deposition was characterised by elemental analysis and X-ray diffractometry (XRD). In vitro assays were carried out to quantify the effects of HA coating on human keratocyte adhesion. Cellular cytokine production was used to assess inflammatory potential. HA coating significantly increased keratocyte adhesion to the carbon matrix (p<0.01). The materials did not induce excessive cytokine production by the adherent keratocytes. In addition, the matrices themselves adsorbed significant levels of the cytokine IL-8 (p<0.05). The results indicate that HA coated carbon matrices provide a suitable environment to enhance in-growth of corneal cells without inducing further inflammation. The materials may also suppress excessive inflammation by adsorption of the cytokine IL-8 into the porous, internal carbon structure. PMID- 19299006 TI - Body-mass index and cause-specific mortality in 900 000 adults: collaborative analyses of 57 prospective studies. AB - BACKGROUND: The main associations of body-mass index (BMI) with overall and cause specific mortality can best be assessed by long-term prospective follow-up of large numbers of people. The Prospective Studies Collaboration aimed to investigate these associations by sharing data from many studies. METHODS: Collaborative analyses were undertaken of baseline BMI versus mortality in 57 prospective studies with 894 576 participants, mostly in western Europe and North America (61% [n=541 452] male, mean recruitment age 46 [SD 11] years, median recruitment year 1979 [IQR 1975-85], mean BMI 25 [SD 4] kg/m(2)). The analyses were adjusted for age, sex, smoking status, and study. To limit reverse causality, the first 5 years of follow-up were excluded, leaving 66 552 deaths of known cause during a mean of 8 (SD 6) further years of follow-up (mean age at death 67 [SD 10] years): 30 416 vascular; 2070 diabetic, renal or hepatic; 22 592 neoplastic; 3770 respiratory; 7704 other. FINDINGS: In both sexes, mortality was lowest at about 22.5-25 kg/m(2). Above this range, positive associations were recorded for several specific causes and inverse associations for none, the absolute excess risks for higher BMI and smoking were roughly additive, and each 5 kg/m(2) higher BMI was on average associated with about 30% higher overall mortality (hazard ratio per 5 kg/m(2) [HR] 1.29 [95% CI 1.27-1.32]): 40% for vascular mortality (HR 1.41 [1.37-1.45]); 60-120% for diabetic, renal, and hepatic mortality (HRs 2.16 [1.89-2.46], 1.59 [1.27-1.99], and 1.82 [1.59-2.09], respectively); 10% for neoplastic mortality (HR 1.10 [1.06-1.15]); and 20% for respiratory and for all other mortality (HRs 1.20 [1.07-1.34] and 1.20 [1.16 1.25], respectively). Below the range 22.5-25 kg/m(2), BMI was associated inversely with overall mortality, mainly because of strong inverse associations with respiratory disease and lung cancer. These inverse associations were much stronger for smokers than for non-smokers, despite cigarette consumption per smoker varying little with BMI. INTERPRETATION: Although other anthropometric measures (eg, waist circumference, waist-to-hip ratio) could well add extra information to BMI, and BMI to them, BMI is in itself a strong predictor of overall mortality both above and below the apparent optimum of about 22.5-25 kg/m(2). The progressive excess mortality above this range is due mainly to vascular disease and is probably largely causal. At 30-35 kg/m(2), median survival is reduced by 2-4 years; at 40-45 kg/m(2), it is reduced by 8-10 years (which is comparable with the effects of smoking). The definite excess mortality below 22.5 kg/m(2) is due mainly to smoking-related diseases, and is not fully explained. PMID- 19299008 TI - Development of pH-responsive chitosan/heparin nanoparticles for stomach-specific anti-Helicobacter pylori therapy. AB - The microorganism now known as Helicobacter pylori is considered to be an important factor in the etiology of peptic ulcers. It can secrete urease enzyme and buffer gastric acids to survive in the stomach. H. pylori can colonize the gastric mucosa and preferentially adheres near the cell-cell junctions of the gastric mucous cells. In this study, pH-responsive nanoparticles were produced instantaneously upon the addition of heparin solution to a chitosan solution with magnetic stirring at room temperature. The nanoparticles appeared to have a particle size of 130-300 nm, with a positive surface charge, and were stable at pH 1.2-2.5, allowing them to protect an incorporated drug from destructive gastric acids. We also demonstrated that the prepared nanoparticles can adhere to and infiltrate cell-cell junctions and interact locally with H. pylori infection sites in intercellular spaces. PMID- 19299009 TI - Composite polymer systems with control of local substrate elasticity and their effect on cytoskeletal and morphological characteristics of adherent cells. AB - At the interface between extracellular substrates and biological materials, substrate elasticity strongly influences cell morphology and function. The associated biological ramifications comprise a diversity of critical responses including apoptosis, differentiation, and motility, which can affect medical devices such as stents. The interactions of the extracellular environment with the substrate are also affected by local properties wherein cells sense and respond to different physical inputs. To investigate the effects of having localized elasticity control of substrate microenvironments on cell response, we have developed a method to control material interface interactions with cells by dictating local substrate elasticity. This system is created by generating a composite material system with alternating, linear regions of polymers that have distinct stiffness characteristics. This approach was used to examine cytoskeletal and morphological changes in NIH 3T3 fibroblasts with emphasis on both local and global properties, noting that cells sense and respond to distinct material elasticities. Isolated cells sense and respond to these local differences in substrate elasticity by extending processes along the interface. Also, cells grown on softer elastic regions at higher densities (in contact with each other) have a higher projected area than isolated cells. Furthermore, when using chemical agents such as cytochalasin-D to disrupt the actin cytoskeleton, there is a significant increase in projected area for cells cultured on softer elastic regions This method has the potential to promote understanding of biomaterial-affected responses in a diversity of areas including morphogenesis, mechanotransduction, stents, and stem cell differentiation. PMID- 19299010 TI - Magnetically targeted thrombolysis with recombinant tissue plasminogen activator bound to polyacrylic acid-coated nanoparticles. AB - We investigated the feasibility and efficacy of target thrombolysis with recombinant tissue plasminogen activator (rtPA) covalently bound to magnetic nanoparticle (MNP) and retained to the target site in vivo by an external magnet. Polyacrylic acid-coated magnetite (PAA-MNP, 246 nm) was synthesized and characterized; rtPA was immobilized to PAA-MNP through carbodiimide-mediated amide bond formation. The enzyme activities of the bound rtPA, as measured by a chromogenic substrate assay and (125)I-fibrinolysis assay, were 87+/-1% and 86+/ 3% of that of free rtPA. Under guidance with the magnet moving back and forth along the iliac artery, the thrombolytic activity of PAA-MNP-rtPA with rtPA equivalent to 0.2mg/kg was determined by flowmetry in a rat embolic model. Intra arterial administration of PAA-MNP-rtPA restored the iliac blood flow within 75 min to 82% of that before the clot lodging, whereas equivalent amount of PAA-MNP or free rtPA exerted no improvement on hemodynamics. At the end of 2-h period, PAA-MNP-rtPA did not alter levels of hemoglobin, hematocrit, or blood cell count. In conclusion, immobilization of rtPA to PAA-MNP with covalent binding resulted in a stable rtPA preparation and predictable amount of rtPA around the target site under magnetic guidance; this approach may achieve reproducible and effective target thrombolysis with <20% of a regular dose of rtPA. PMID- 19299011 TI - Development of polyion complex micelles for encapsulating and delivering amphotericin B. AB - A block copolymer poly(2-ethyl-2-oxazoline)-block-poly(aspartic acid) (PEOz-b PAsp) was synthesized and investigated as the carrier of antifungal drug amphotericin B (AmB). Polyion complex (PIC) micelles with clear core-shell structures were identified by TEM, which revealed that the PAsp segment became hydrophobic after it interacted with AmB. PEOz-b-PAsp increased not only the solubility of AmB but also simultaneously the drug potency. The prolonged release of AmB from micelles effectively inhibited the growth of Candida albicans even after three days of administration. Moreover, the in vitro cytotoxicity of AmB loaded micelles was less than that of Fungizone, which is a powerful antifungal antibiotic that is adopted to treat various fungal infections. The PEOz-b-PAsp PIC micelles with lower cytotoxicity and higher potency than Fungizone represent a potential means of encapsulating basic/amphoteric drugs. PMID- 19299012 TI - Poly(lactide)-vitamin E derivative/montmorillonite nanoparticle formulations for the oral delivery of Docetaxel. AB - Four systems of nanoparticles of biodegradable polymers were developed in this research for oral delivery of anticancer drugs with Docetaxel used as a model drug, which include the poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs), the poly(lactide)-vitamin E TPGS nanoparticles (PLA-TPGS NPs), the poly(lactic-co glycolic acid)-montmorillonite nanoparticles (PLGA/MMT NPs) and the poly(lactide) vitamin E TPGS/montmorillonite nanoparticles (PLA-TPGS/MMT NPs). Vitamin E TPGS stands for d-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS), which is a water-soluble derivative of natural vitamin E formed by esterification of vitamin E succinate with polyethylene glycol (PEG) 1000. The design was made to take advantages of TPGS in nanoparticle technology such as high emulsification effects and high drug encapsulation efficiency, and those in drug formulation such as high cellular adhesion and adsorption. MMT of similar effects is also a detoxifier, which may cure some side effects caused by the formulated drug. The drug-loaded NPs were prepared by a modified solvent extraction/evaporation method and then characterized for their MMT content, size and size distribution, surface charge and morphology, physical status and encapsulation efficiency of the drug in the NPs, and in vitro drug release profile. Cellular uptake of the coumarin 6 loaded NPs was investigated. In vitro cancer cell viability experiment showed that judged by IC(50), the PLA-TPGS/MMT NP formulation was found 2.89, 3.98, 2.12 fold more effective and the PLA-TPGS NP formulation could be 1.774, 2.58, 1.58 fold more effective than the Taxotere((R)) after 24, 48, 72h treatment, respectively. In vivo PK experiment with SD rats showed that oral administration of the PLA-TPGS/MMT NP formulation and the PLA-TPGS NP formulation could achieve 26.4 and 20.6 times longer half-life respectively than i.v. administration of Taxotere((R)) at the same 10mg/kg dose. One dose oral administration of the NP formulations could realize almost 3 week sustained chemotherapy in comparison of 22h of i.v. administration of Taxotere((R)). The oral bioavailability can be enhanced from 3.59% for Taxotere((R)) to 78% for the PLA-TPGS/MMT NP formulation and 91% for the PLA-TPGS NP formulation respectively. Oral chemotherapy by nanoparticles of biodegradable polymers is feasible. PMID- 19299013 TI - The effect of the co-immobilization of human osteoprogenitors and endothelial cells within alginate microspheres on mineralization in a bone defect. AB - Bone regeneration seems to be dependant on cell communication between osteogenic and endothelial cells arising from surrounding blood vessels. This study aims to determine whether endothelial cells can regulate the osteogenic potential of osteoprogenitor cells in vitro and in vivo, in a long bone defect, when co immobilized in alginate microspheres. Alginate is a natural polymer widely used as a biomaterial for cell encapsulation. Human osteoprogenitors (HOP) from bone marrow mesenchymal stem cells were immobilized alone or together with human umbilical vein endothelial cells (HUVEC) inside irradiated, oxidized and RGD grafted alginate microspheres. Immobilized cells were cultured in dynamic conditions and cell metabolic activity increased during three weeks. The gene expression of alkaline phosphatase and osteocalcin, both specific markers of the osteoblastic phenotype, and mineralization deposits were upregulated in co immobilized HOPs and HUVECs, comparing to the immobilization of monocultures. VEGF secretion was also increased when HOPs were co-immobilized with HUVECs. Microspheres containing co-cultures were further implanted in a bone defect and bone formation was analysed by muCT and histology at 3 and 6 weeks post implantation. Mineralization was observed inside and around the implanted microspheres containing the immobilized cells. However, when HOPs were co immobilized with HUVECs, mineralization significantly increased. These findings demonstrate that co-immobilization of osteogenic and endothelial cells within RGD grafted alginate microspheres provides a promising strategy for bone tissue engineering. PMID- 19299014 TI - siRNA-mediated down-regulation of iASPP promotes apoptosis induced by etoposide and daunorubicin in leukemia cells expressing wild-type p53. AB - Oncoprotein inhibitory member of the ASPP family (iASPP) is a key inhibitor of tumor suppressor p53. Our previous study revealed that the expression of iASPP in acute leukemia (AL) patients was higher than that of normal control which implied that iASPP might play an important role in the pathogenesis and/or disease progression of AL. In this study, the iASPP expression was blocked by RNA interference (RNAi) in two leukemic cell lines, Nalm6 and K562, to explore the effects of iASPP on leukemia cells. The results indicated that down-regulation of endogenous iASPP increased p53-dependent apoptosis of leukemia cells. Thus, iASPP could be a molecular target in leukemia therapy. PMID- 19299015 TI - A comparison of mental health legislation from diverse Commonwealth jurisdictions. AB - INTRODUCTION: In the regulation of involuntary treatment, a balance must be found between duties of care and protection and the right to self-determination. Despite its shared common roots, the mental health legislation of Commonwealth countries approaches this balance in different ways. When reform is planned, lessons can be learned from the experiences of other countries. METHOD: Criteria for involuntary treatment used in a sample of 32 Commonwealth Mental Health Acts were compared using a framework developed from standards derived from the Universal Declaration of Human Rights. Reasons for non-compliance were considered and examples of good practice were noted. Changes in the criteria used over time and across areas with differing levels of economic development were analysed. RESULTS: 1. Widespread deviation from standards was demonstrated, suggesting that some current legislation may be inadequate for the protection of the human rights of people with mental disorders. 2. Current trends in Commonwealth mental health law reform include a move towards broad diagnostic criteria, use of capacity and treatability tests, treatment in the interests of health rather than safety, and regular reviews of treatment orders. Nevertheless, there are some striking exceptions. DISCUSSION: Explanations for deviation from the standards include differing value perspectives underpinning approaches to balancing conflicting principles, failure to keep pace with changing attitudes to mental disorder, and variations in the resources available for providing treatment and undertaking law reform. Current good practice provides examples of ways of dealing with some of these difficulties. PMID- 19299016 TI - Psychologists abandon the Nuremberg ethic: concerns for detainee interrogations. AB - In the aftermath of 9-11, the American Psychological Association, one of the largest U.S. health professions, changed its ethics code so that it now runs counter to the Nuremberg Ethic. This historic post-9-11 change allows psychologists to set aside their ethical responsibilities whenever they are in irreconcilable conflict with military orders, governmental regulations, national and local laws, and other forms of governing legal authority. This article discusses the history, wording, rationale, and implications of the ethical standard that U.S. psychologists adopted 7 years ago, particularly in light of concerns over health care professionals' involvement in detainee interrogations and the controversy over psychologists' prominent involvement in settings like the Guantanamo Bay Detainment Camp and the Abu Ghraib prison. It discusses possible approaches to the complex dilemmas arising when ethical responsibilities conflict with laws, regulations, or other governing legal authority. PMID- 19299017 TI - Making sense of 'consent' in a constrained environment. AB - BACKGROUND: This project investigates patients' and practitioners' experiences and understandings of the consent process, as it is governed by the Mental Health Act in Great Britain. AIMS: We aim to illuminate our respondents' experiences of the consent process, and to explore their attempts to make sense of that process. METHOD: Semi-structured interviews with 5 Responsible Medical Officers, and 7 of their consenting adult patients, were conducted at a medium-secure psychiatric hospital. We approached the analysis from the perspective of Interpretative Phenomenological Analysis. RESULTS: Our analysis begins with an account of some of the common phenomenological consequences of the consent process as our participants understand them, but then moves on to discuss some of the contextual constraints which are evident from their negotiation of these understandings. CONCLUSIONS: We conclude by outlining a number of emergent issues relevant to the current development of new Mental Health legislation. These include: mechanisms to allow collaboration with user groups; a more consultative role for users in their own treatment decisions; formal training and support for those conducting competency assessments; and more flexible and transparent legislative frameworks. PMID- 19299018 TI - Triclosan is a potent inhibitor of estradiol and estrone sulfonation in sheep placenta. AB - The personal care product Triclosan, 5-chloro-2(2,4-dichlorophenoxy)-phenol, is widely used in consumer products as an antibacterial agent and is increasingly found in the environment as a contaminant of sewage sludge and wastewater. This compound has been identified in plasma and urine of people in the United States, Sweden and Australia. Triclosan is known to inhibit sulfonation of phenolic xenobiotics and is structurally related to inhibitors of estrogen sulfotransferase, such as polychlorobiphenylols. In pregnancy, the placenta is an important source of estrogen, which is needed for normal fetal development and successful parturition, and estrogen sulfotransferase is thought to play an important role in regulation of estrogen availability. In this study, we examined the effect of Triclosan on sheep placental cytosolic sulfotransferase activity with 17-beta-estradiol and estrone as substrates. For comparison, we studied the effects of 4-hydroxy-3,3',4',5-tetrachlorobiphenyl and 2'-hydroxytriclocarban on estradiol sulfonation. The apparent K(m) for placental cytosolic sulfotransferase activity with estradiol as substrate was 0.27 +/- 0.06 nM (mean +/- S.D., n = 3 individuals) and with estrone as substrate was 1.86 +/- 0.22 nM. Partial substrate inhibition was observed with estradiol at concentrations higher than 10 20 nM, as is typical of estrogen sulfotransferases (SULT1E1) in other species. Studies of the effect of Triclosan on estrogen sulfotransferase activity were conducted with several concentrations (0.1-6 nM) of estradiol and with 2 nM estrone. Triclosan was a very potent inhibitor of both estradiol and estrone sulfonation. For estradiol the inhibition was shown to be mixed competitive/uncompetitive, with K(ic) of 0.09 +/- 0.01 nM and K(iu) of 5.2 +/- 2.9 nM. The IC(50) for inhibition of estrone sulfonation was 0.60 +/- 0.06 nM. At an environmentally relevant concentration of 1 uM, Triclosan was not a substrate for glucuronidation in sheep placental microsomes. Triclosan could be sulfonated in placental cytosol with K(m) 1.14 +/- 0.18 uM and V(max) 160 +/- 26 pmol/min/mg protein, however the calculated rates of Triclosan sulfonation were negligible at the low nM concentrations that potently inhibit estrogen sulfonation. The high potency of Triclosan as an inhibitor of estrogen sulfotransferase activity raises concern about its possible effects on the ability of the placenta to supply estrogen to the fetus, and in turn on fetal growth and development. PMID- 19299019 TI - STAT3 tyrosine phosphorylation is critical for interleukin 1 beta and interleukin 6 production in response to lipopolysaccharide and live bacteria. AB - Both interleukin 1 beta (IL-1beta) and interleukin-6 (IL-6) are pro-inflammatory cytokines that play a major role in inflammatory diseases as well as cancer. In this work we investigated the signaling pathway involving lipopolysaccharide (LPS)-mediated IL-1beta and IL-6 production in murine macrophage cell lines and primary macrophages. We show that in response to LPS, the JAK/STAT pathway is activated, leading to tyrosine phosphorylation at residue 705 on STAT3 and at residue 701 on STAT1, respectively. A newly developed STAT3 specific inhibitor (stattic) blocked LPS-mediated STAT3 tyrosine phosphorylation and led to inhibition of LPS-mediated IL-1beta and IL-6 production but not TNF-alpha production. Knockdown of STAT3 expression via small interfering RNA (siRNA) decreased the level of STAT3 expression in Raw 264.7 cells and decreased STAT3 tyrosine phosphorylation in response to LPS treatment. Quantitative real time PCR and Western analysis of cells treated with inhibitor or STAT3 siRNA after LPS treatment showed a significant reduction of IL-1beta and IL-6 mRNA and protein compared to cells treated with LPS alone. Moreover stattic abrogated IL-1beta formation in response to extracellular bacteria Staphylococcus aureus and Escherichia coli in murine peritoneal macrophages. This inhibition did not affect caspase-1 activation. These results highlight the complex role of STAT3 in cytokine production and the key role of STAT3 tyrosine phosphorylation in IL 1beta and IL-6 production in response to inflammation. PMID- 19299020 TI - The immunoglobulin heavy chain locus in the reptile Anolis carolinensis. AB - We describe the entire immunoglobulin heavy chain (IgH) locus from the reptile Anolis carolinensis. The heavy chain constant (C(H)) region includes C mu, C delta and C upsilon genes. This is the first description of a C upsilon gene in the reptilian class. Variable (V(H)), diversity (D(H)) and joining (J(H)) genes are located 5' from the constant (C(H)) chain complex locus. The C mu and C upsilon genes encode antibodies with four immunoglobulin domains. The C delta gene encoded an 11 domain delta heavy chain as in Eublepharis macularius. Seventy V(H) genes, belonging to 28 families, were identified, and they can be sorted into five broader groups. The similarity of the organization of the reptilian genes with those of amphibians and mammals suggests the existence of a process of heavy chain genomic reorganization before the radiation of tetrapod vertebrates. PMID- 19299021 TI - Expression of interleukins-23 and 27 leads to successful gene therapy of hepatocellular carcinoma. AB - IL-23 and IL-27 are two novel IL-12 cytokine family members who are quite similar to, but yet clearly distinct from IL-12 in their structures and T-cell stimulatory mechanisms. Here, we demonstrated that either IL-27 or IL-23 has potent antitumor activity in murine models of MM45T.Li hepatocellular carcinoma (HCC). These potent antitumor effects were induced primarily by CD8(+)T cells, secreting IFN-gamma while CD4(+)T cells were also involved as a help of antitumor immunity. However, the antitumor response induced by IL-27 was observed from an early stage of tumor growth whereas that of IL-23 was only evident in the late stage of tumor cell proliferation. IL-23 could induce mice to develop a long-term systemic immunologic memory response against parental MM45T.Li tumors cells, an effect IL-27 was not able to accomplish. CTLs specific for MM45T.Li cells were significantly induced by IL-23, whereas antitumor efficacy mediated by IL-27 and IL-12 involved NK cells, which IL-23 failed to activate. Furthermore, we demonstrated that CD40 expression also plays an important role in the induction of antitumor activities by IL-27, IL-23 or IL-12. Together our data suggest that IL-27 and IL-23 may be two novel and attractive candidate agents to apply to cancer immunotherapy. PMID- 19299022 TI - Interleukin-10 promoter polymorphisms in myasthenia gravis. AB - Interleukin 10 (IL-10) is secreted by several hemopoietic cells and suppresses the Th1 mediated immune response, while stimulating B cell differentiation and the humoral immune response. IL-10 expression in Con A-stimulated peripheral blood mononuclear cells is related to three polymorphisms in the promoter region of the IL-10 gene; G/A at position -1082, T/C at position -819 and A/C at position -592. We analyzed the distribution of these IL-10 polymorphisms in 64 MG patients and 87 healthy blood donors to determine any influence on MG susceptibility. MG patients had a significantly higher frequency of the ACC/ACC haplotype (12.5% vs 3.4% in controls), as had the subgroups with late onset MG and thymomatous MG (20.0% and 21.4%, respectively). Early onset MG patients had a high frequency of the ATA/ATA haplotype (19.2% vs 3.4% in controls). Titin Ab positive MG patients had high ACC/ACC (20.0%). This study indicates a direct link between IL-10 and MG pathogenesis, although the complex role of this multi faceted cytokine in vivo is as yet not fully elucidated. PMID- 19299023 TI - Identification of novel variants in the Myosin VIIA gene of patients with nonsyndromic hearing loss from Taiwan. AB - OBJECTIVE: To determine whether variants of exons 7, 11, 22 and 28 of the MYO7A gene are causes of nonsyndromic deafness in Taiwanese. METHODS: We screened a total of 331 unrelated Taiwanese individuals (age range, 4-22 years), including 231 patients with severe to profound nonsyndromic hearing loss and 100 individuals with normal hearing. Genomic DNA was extracted from peripheral blood leukocytes and then subjected to PCR to amplify selected exons and flanking introns of the MYO7A gene; the amplified products were screened for base mutations by autosequence. Data from the two groups were then compared using the chi-square (chi(2)) test. RESULTS: The analysis revealed six variants in 3 out of 4 screened exons and flanking intronic sequences of the MYO7A gene (exons 7, 11, and 22). Three missense variants were found only in patients with hearing loss and were heterozygous, including Arg206Cys, Arg206His and Thr381Met. A variant, c.IVS22+58G>A, was found in intron 22 of the MYO7A gene from both patients and control group. Allele frequencies of c.IVS22+58G>A were shown to be significant between the two groups using chi(2) test (P<0.05). CONCLUSION: Our results indicate that Arg206 and Thr381 residues in the motor head region of MYO7A protein are critical sites and the mutations of these residues may lead to the development of nonsyndromic deafness. PMID- 19299024 TI - Estrogen-induced plasticity from cells to circuits: predictions for cognitive function. AB - Controversy regarding estrogen action in the brain remains at the forefront of basic, translational and clinical science for women's health. Here, I provide an integrative analysis of estrogen-inducible plasticity and posit it as a strategy for predicting cognitive domains affected by estrogen in addition to sources of variability. Estrogen enhancement of plasticity is evidenced by increases in neurogenesis, neural network connectivity and synaptic transmission. In parallel, estrogen increases glucose transport, aerobic glycolysis and mitochondrial function to provide the ATP necessary to sustain increased energetic demand. The pattern of plasticity predicts that estrogen would preferentially affect cognitive tasks of greater complexity, temporal demand and associative challenge. Thus, estrogen deprivation should be associated with decrements in these functions. Estrogen regulation of plasticity and bioenergetics provides a framework for predicting estrogen-dependent cognitive functions while also identifying sources of variability and potential biomarkers for identifying women appropriate for hormone therapy. PMID- 19299025 TI - Variability in the subcellular distribution of ion channels increases neuronal diversity. AB - The exact location of an ion channel on the axo-somato-dendritic surface of a nerve cell crucially affects its functional impact. Recent high-resolution immunolocalization experiments examining the distribution of GABA and glutamate receptors, voltage-gated potassium and sodium channels and hyperpolarization activated mixed cation (HCN) channels clearly demonstrate the lack of simple rules concerning their subcellular distribution. For example, the density of HCN1 subunits in pyramidal cells increases 60-fold from soma to distal dendrites but is uniform over the somato-dendritic surface of olfactory bulb external tufted cells and is highest in the axon of cortical and cerebellar basket cells. Such findings highlight the necessity of determining the precise subcellular location and density of each ion channel in every cell type. Here, I suggest that variations in the subcellular distribution of ion channels are previously unrecognized means of increasing neuronal diversity and, thus, the computational power of the brain. PMID- 19299026 TI - Serial imaging changes during treatment of Takayasu arteritis with pulmonary artery stenosis. AB - Most cases of chronic stenosis or occlusive lesions of the pulmonary arteries are attributed to thromboembolism, and pulmonary arteritis is extremely rare as the primary cause of these entities. We report a case of pulmonary stenosis and occlusion caused by Takayasu arteritis. The patient was a 54-year-old woman who presented with dyspnea. Total occlusion of the left pulmonary artery and significant stenosis of the right pulmonary artery caused by Takayasu arteritis were confirmed by various imaging modalities including pulmonary angiography, 18fluorodeoxyglucose-positron emission tomography, magnetic resonance imaging and real-time three-dimensional transesophageal echocardiography. After 6 weeks of steroid therapy, follow-up imaging studies showed that the stenotic lesion had resolved. PMID- 19299027 TI - GLP-1 receptor agonists: targeting both hyperglycaemia and disease processes in diabetes. PMID- 19299028 TI - Portal hypertension and development of hepatocellular carcinoma: factors influencing significance in prognostic models. PMID- 19299029 TI - Association of serum uric acid level with non-alcoholic fatty liver disease: a cross-sectional study. AB - BACKGROUND/AIM: Serum uric acid level has been suggested to be associated with factors that contribute to the metabolic syndrome. The aim of this study was to investigate the association of serum uric acid level with non-alcoholic fatty liver disease (NAFLD). METHODS: A cross-sectional study was performed among the employees of Zhenhai Refining & Chemical Company Ltd., Ningbo, China. RESULTS: The study included 8925 subjects (6008 men) with a mean age of 43 years. The prevalence rates of NAFLD and hyperuricemia were 11.78% and 14.71%, respectively. NAFLD patients had significantly higher serum uric acid levels than controls (370.3+/-86.6 vs. 321.1+/-82.6 micromol/L; P<0.001). The prevalence rate of NAFLD was significantly higher in subjects with hyperuricemia than in those without hyperuricemia (24.75% vs. 9.54%; P<0.001), and the prevalence rate increased with progressively higher serum uric acid levels (P value for trend <0.001). Multiple regression analysis showed that hyperuricemia was associated with an increased risk of NAFLD (odds ratio [OR]: 1.291, 95% confidence interval [CI]: 1.067-1.564; P<0.001). CONCLUSION: Serum uric acid level is significantly associated with NAFLD, and elevated serum uric acid level is an independent risk factor for NAFLD. PMID- 19299030 TI - Differences in general practice initiated expenditures across Danish local health authorities-A multilevel analysis. AB - OBJECTIVE: To assess differences in expenditures of general practice services across local health authorities (counties). METHODS: A multilevel analysis of 2123 general practices nested within 15 counties is used to assess between county variations in general practice expenditures in year 2006. Schmidt and Sickles' fixed effect efficiency estimator is used to assess the potential for reducing expenditures (fee-for-service, prescriptions, and referrals). RESULTS: Expenditures associated with general practice constitute 38% of total health care expenditures. 42% of the variation in these expenditures is attributable to geographical location (county). Mean efficiency is estimated to 87% corresponding to a savings potential of 700 DKK per insured person. Referrals to specialised care are the main source of variation in GP initiated expenditures across the counties. Expenditures associated with out-patient referrals and referrals to practicing specialist are negatively correlated (p<0.01) indicating a substitution effect. CONCLUSIONS: Our results indicate that primary care reforms aiming at reducing GP initiated expenditures should focus on general practice access to low level specialised care rather than reforming GP remuneration systems. PMID- 19299031 TI - Adenocarcinoma has an excellent outcome with pemetrexed treatment in Korean patients: a prospective, multicenter trial. AB - OBJECTIVE: This prospective multicenter study conducted by the Korean Cancer Study Group evaluated the efficacy and safety of pemetrexed in Korean patients with advanced non-small cell lung cancer (NSCLC) who had prior chemotherapy. PATIENTS AND METHODS: Patients with stage IIIB or IV NSCLC in whom prior chemotherapy failed received pemetrexed 500 mg/m(2) every 3 weeks with folic acid and vitamin B12 supplementation until disease progression or the development of intolerable toxicity. Eighty-one patients were enrolled. RESULTS: The overall response rate for 78 evaluable patients was 5.1% [95% confidence interval (CI) 1.4-12.6; partial response 4/78, no complete response]. The disease control rate including complete, partial response and stable disease was 46.2% (36/78, 95% CI 34.8-57.8). With a median 8.7 months follow-up, the median time to progression was 3.1 months (95% CI 1.17-5.03) and the median overall survival (OS) was 7.8 months (95% CI 5.19-10.35). The median OS for patients with adenocarcinoma histology was 18.7 months compared to 6.1 months for non-adenocarcinoma. In a multivariate analysis, Eastern Cooperative Oncology Group performance status 0-1 [hazards ratio (HR)=0.331, 95% CI 0.135-0.814] and adenocarcinoma (HR=0.504, 95% CI 0.283-0.899) were independent factors for prolongation of overall survival. CONCLUSIONS: Pemetrexed monotherapy has promising efficacy in patients with advanced NSCLC as a second-line therapy with less hematologic and non-hematologic toxicity, especially in those with adenocarcinoma histology. PMID- 19299032 TI - Sex-associated differences in non-small cell lung cancer in the new era: is gender an independent prognostic factor? AB - BACKGROUND: Women with non-small cell lung cancer (NSCLC) appear to have better survival. This study aimed to evaluate sex differences in NSCLC in recent years. The true effect of gender on the overall survival was analyzed taking other prognostic factors into account. METHODS: A cohort of consecutive NSCLC patients was prospectively enrolled from January 2002 to December 2005, and followed-up until December 2006. They were clinically and pathologically staged and underwent homogenous treatment algorithms. Demographics, histology, and disease stage between sexes were compared. The clinical prognostic factors to be analyzed in addition to gender included stage, age, smoking history and histology. The overall survival of females and males within relevant subgroups defined by smoking history and histology was also compared. RESULTS: Of the 738 patients, 695 were analyzed with a definite stage (94.2%; 315 females and 380 males), which was similar in both sexes. Females were younger (median age: 59.5 years vs. 65.0 years; P<0.001) and more likely to have adenocarcinoma (81% vs. 60.5%; P<0.001). Patients with earlier stage, younger patients, never-smokers and females had better overall survival in univariate analyses and no significant survival difference was noted between adenocarcinoma and squamous cell carcinoma. Multivariate analyses demonstrated age, smoking history and gender to have a hazard ratio 1.46 (95% confidence interval, CI 1.21-1.76; P<0.001), 1.27 (95% CI 0.97-1.65; P=0.082), and 1.18 (95% CI 0.90-1.55; P=0.226), respectively. Subgroup analyses revealed the survival of never-smoker males with adenocarcinoma was similar to that of females. CONCLUSIONS: There are sex-related differences in the clinico-pathologic characteristics and survival of NSCLC patients. The survival advantages of females could be attributed to the younger age and lower smoking prevalence. Never-smokers with adenocarcinoma should be given special attention regardless of sex as they imply better survival with different treatment outcomes. PMID- 19299033 TI - Lung adenocarcinoma as a solitary pulmonary nodule: prognostic determinants of CT, PET, and histopathologic findings. AB - We aimed to retrospectively compare CT, PET, and histopathologic (the extent of bronchioloalveolar carcinoma [BAC] components) findings of solitary pulmonary nodular (SPN) adenocarcinomas of the lung to determine their value as prognostic determinants. We reviewed CT and PET characteristics of tumors and pathologic specimens from 65 consecutive patients who underwent surgical resection for SPN adenocarcinomas. Nodule size and TDR (tumor shadow disappearance rate) were assessed from CT scans, and maximum standardized uptake value (SUVmax) of tumors was measured at PET. On pathologic examination, BAC, non-BAC, and central fibrous scar ratios were quantified. Prognosis was evaluated by noting disease recurrence during a minimum 12-month follow-up period after curative resection. The interrelationships between TDR, SUVmax, BAC, and non-BAC ratio were studied, and relationships between recurrence and various variables were analyzed. The median follow-up time was 33 months, and seven patients (11%) developed disease recurrence after surgical resection. TDR at CT and SUVmax at PET correlated well with pathologic BAC and non-BAC ratios. Between subgroups with and without recurrence, there were significant differences in SUVmax and BAC and non-BAC ratios. Based on univariate survival analyses, pathologic BAC and non-BAC ratios were risk factors significantly related to recurrence, but only high non-BAC ratio remained as an independent factor associated with recurrence in the multivariate analysis (hazard ratio [HR]=0.956, P=0.013). Among the factors examined, pathologic non-BAC ratio is the only independent risk factor for poor prognosis in patients with SPN adenocarcinomas. PMID- 19299035 TI - Patriarchy, masculinities and health inequalities. PMID- 19299036 TI - [Social determinants vs. lifestyle in type 2 diabetes mellitus in Andalusia (Spain): difficulty in making ends meet or obesity?]. AB - OBJECTIVE: To assess the relationship between difficulty in making ends meet and obesity and type 2 diabetes mellitus in Andalusia. METHODS: We performed a cross sectional survey based on the Andalusian Health Survey-2003. Measures of the prevalence and odds ratios (OR) of type 2 diabetes mellitus were calculated according to lifestyle variables, obesity and type 2 diabetes mellitus according to self-perceived difficulties in making ends meet. RESULTS: The age-adjusted OR of type 2 diabetes mellitus in obese individuals with respect to those with normal weight was 2.52 (95%CI: 1.63-3.88) in women and 2.13 (95%CI: 1.28-3.54) in men. A sedentary lifestyle was not related to a significant risk of type 2 diabetes mellitus in either sex. For women with greater difficulties in making ends meet compared with women with less difficulty, the risk of being obese, adjusted by age and physical exercise, was 3.03 (95%CI: 1.96-4.66), and the risk of having type 2 diabetes mellitus, adjusted by age, physical exercise and body mass index, was 2.55 (95%CI: 1.28-5.10). In men, none of the estimated OR was statistically significant. CONCLUSION: For women, difficulties in making ends meet reflect a socio-economic context that contributes to a greater risk of obesity and type 2 diabetes mellitus. The gender and social inequalities identified could help to broaden the current framework of risks for type 2 diabetes mellitus, which is excessively focused on individual behaviors. Self perceived difficulties in making ends meet could be useful as a poverty indicator in the study of inequalities in health. PMID- 19299037 TI - Novel topoisomerase I-targeting antitumor agents synthesized from the N,N,N trimethylammonium derivative of ARC-111, 5H-2,3-dimethoxy-8,9-methylenedioxy-5 [(2-N,N,N-trimethylammonium)ethyl]dibenzo[c,h][1,6]naphthyridin-6-one iodide. AB - Several new TOP1-targeting agents were prepared using as an intermediate the N,N,N-trimethyl quaternary ammonium salt 2 of ARC-111. Direct displacement of the quaternary ammonium group with hydroxide, cyclopropylamine, imidazole, 1H-1,2,3 triazole, alkylethylenediamines, ethanolamine, and polyhydroxylated alkylamines provides a convenient means for furthering insight into the structure-activity relationships within this series of non-camptothecin TOP1-targeting agents. The relative TOP1-targeting activities and cytotoxicities were evaluated in RPMI8402 and P388 cells and their camptothecin-resistant variants. Their potential to serve as substrates for the efflux transporters MDR1 and BCRP, which are associated with multidrug resistance, was also assessed. PMID- 19299038 TI - Synthesis and in vitro anticancer activity of 2,4-azolidinedione-acetic acids derivatives. AB - The synthesis and evaluation of anticancer activity of 2,4 thia(imida)zolidinedione-3- and 5-acetic acids amides were described. The structures of compounds were determined by IR, (1)H NMR, and MS analysis. In vitro anticancer activity of these compounds has been tested in National Cancer Institute (NCI) and the relationships between structure and anticancer activity are discussed. Among 2,4-azolidinedione-acetic acids derivatives 2-[5-(4 chlorobenzylidene)-2,4-dioxo-imidazolidin-3-yl]-N-(2-trifluoromethyl-phenyl) acetamide (Ic) was superior to other related compounds in terms of high selectivity for the leukemia CCRF-CEM (logGI(50)=-6.06), HL-60(TB) (logGI(50)= 6.53), MOLT-4 (logGI(50)=-6.52) and SR (logGI(50)=-6.51) cell lines. PMID- 19299039 TI - [A restrictive cardiac insufficiency]. PMID- 19299040 TI - [Abdominal pain and purpura]. PMID- 19299041 TI - [New drugs in oncology and skin toxicity]. AB - The targeted agents have considerably modified the therapeutic approach of cancer over the last few years. The use of these new agents has been associated with the occurrence of new side-effects among which cutaneous side-effects are the most prominent. Although they rarely compromise the vital prognosis, these cutaneous side-effects must be taken into consideration in order to improve treatment compliance and to maintain an acceptable quality of life. Accurate identification of these cutaneous side-effects is therefore critical to improve the management of these patients. A better understanding of the mechanisms underlying cutaneous signs is also an important issue as it gives us the opportunity to increase our knowledge of the skin pathophysiology. Furthermore, the cutaneous manifestations could sometimes be associated to the antitumor response. The skin is an easily accessible interface, allowing addressing the complexity of the targeted therapies effect on tissues. PMID- 19299042 TI - [Anticoagulation clinics for outpatients: a 5-year experience]. AB - INTRODUCTION: Anticoagulation clinics and computerized management of chronic oral anticoagulation increase the time spent in the therapeutic range with both mortality and morbidity reduction. Usually, anticoagulation clinics are hospital based medical care centers. We report the five-year results from a general medicine center (CSCTA) using a computer-assisted management. METHODS: A prospective cohort observational study of 530 primary care patients that were receiving long term oral anticoagulation. RESULTS: Cardiac arrhythmia (55%), heart valve disease and venous thrombo-embolic disease (30%) represented the most common indications of oral anticoagulation. Patients received fluindione, warfarin and acenocoumarol in 80%, 13% and 7%, respectively. The duration of treatment was at least one year in 54% of the cases, and was at least three years in 25% of the cases. The rate of patients that were in average within the therapeutic range (INR 2-3) was 72%, while 12% were under and 16% over the therapeutic range. Corresponding rates were 82, 17 and 1% respectively for all anticoagulation targets (INR 1.5-4.5). Twenty-six bleeding events (4.9 per 100 patient-years) and four thrombotic complications (0.75 per 100 patient-years) occurred. Life-threatening hemorrhage occurred in 1.3 per 100 patient-years. After the equilibration of the anticoagulation, the average delay of control between two consecutive INR was 19 days. CONCLUSION: The results obtained with CSCTA were similar to those reported by other anticoagulation clinics regarding hemorrhagic complications and time spent in the therapeutic range. In contrast, thrombotic events were less frequent. Because of the absence of a control group, a medico-economic analysis could not be performed. PMID- 19299043 TI - [Acute renal failure and rhabdomyolysis secondary to prolonged hunger strike. A case report]. AB - Acute renal failure following hunger strike has been rarely reported. We report a 47-year-old man, prisoner, who developed an acute renal failure secondary to hypovolemia and major rhabdomyolysis. Failure of hydration with persistence of oliguria and secondary pulmonary edema required hemodialysis with eventually a favorable outcome. PMID- 19299044 TI - [Mitochondrial diabetes: clinical features, diagnosis and management]. AB - Mitochondrial diabetes affects up to 1% of patients with diabetes and is often unrecognised by the physicians. Maternally inherited diabetes and deafness (MIDD) resulting from the mutation 3243A>G of the mitochondrial DNA is the most frequent mutation associated with mitochondrial diabetes. This review summarizes the range of clinical phenotypes associated with MIDD and outlines the advances in genetic diagnosis, pathogenesis and management of these patients. PMID- 19299045 TI - [Antidepressant use in demented elderly subjects: current data]. AB - Depression and behavioural and psychological symptoms of dementia (BPSD) have a significant impact on the worsening of dementia because they increase the cognitive and functional decline and they have a significant impact on the vital prognosis. Physicians should be particularly careful in the use of antidepressants in the elderly, particularly in the frail elderly. Indeed, most studies have included patients aged at least 65 years without frailty criteria, but rarely those aged over 75 years and/or frail. As they are used in clinical practice, selective serotonin reuptake inhibitors, which initially appeared to have low risks, have been associated with many and dangerous adverse effects, particularly in elderly subjects. At present, there is a lack of data to assess the benefit-risk ratio of antidepressants in the treatment of depression and BPSD in patients with Alzheimer's disease or other dementias. Among the drugs frequently used in studies in order to evaluate these indications, citalopram and moclobemide are those associated with a low risk of adverse events and a significant effectiveness on depression signs and behavioural and BPSD. It is necessary to assess the effectiveness and adverse effects of antidepressants in demented elderly subjects through several studies. PMID- 19299046 TI - [Takayasu arteritis: a French single centre experience]. AB - Takayasu arteritis (TA) is an uncommon large-vessel arteritis. We report our French single-center experience in the management of patients with TA (Pitie Salpetriere Hospital, Paris). TA is diagnosed in patients presenting with a large vessel arteriopathy, in whom several inflammatory and non-inflammatory differential diagnoses are ruled out by appropriate investigations. Treatment of active disease is primarily based on corticosteroids but other immunosuppressive drugs are frequently needed. Anti-platelets agents, statins and antihypertensive drugs are frequently considered. There is no validated disease activity criterion in TA; thus, we generally consider the disease as being active in the presence of the following criteria: firstly, the presence of constitutional or ischemic symptoms; secondly, the increased acute phase reactants; thirdly, the mural contrast enhancement in CT-scan or fourthly, the mural contrast enhancement or signal abnormalities in MRI; fifthly, the abnormal vascular uptake in PET-scan. When TA is active, our follow-up recommendation is to perform an ultrasonography of the supra-aortic vessels and an aortic MRI or CT-scan twice a year. When TA appears to be inactive, we recommend to perform these investigations once a year. Surgical treatment of TA is limited to a few indications. The overall prognosis of TA is good but the quality of life is altered. Management of TA patients is difficult because of the lack of reliable diagnostic criteria, consensual therapeutic strategies and validated disease activity criteria. Further studies should focus on the pathogenesis of the disease and help define better disease activity criteria. PMID- 19299047 TI - [Euthyroid Graves' disease: a case report]. AB - Euthyroid Graves' disease is defined as an ophtalmopathy without any clinical or biological signs of thyroid dysfunction. It highlights the apparent dissociation between orbitopathy and thyroid disease. Diagnosis is often too late while early treatment could really improve functional prognosis. We report a 57-year-old woman who presented with this entity and that illustrates both the diagnostic difficulty and disease course after intravenous corticosteroid therapy. PMID- 19299048 TI - [Targeted therapies and their indications in solid neoplasias]. AB - Targeted therapies are widely used in cancer because of their effectiveness, even in tumours that are resistant to conventional chemotherapy such as kidney or hepatocellular carcinomas. There are different families classified according to their mode of action. The antiangiogenics block tumor angiogenesis by acting on VEGF or its receptor. The main molecules are bevacizumab, sunitinib, and sorafinib. HER inhibitors work by blocking these receptors, which control different signaling intracellular pathways, and include an inhibitor of HER2, trastuzumab, and various inhibitors of HER1, or EGFR, including cetuximab, erlotinib, and gefitinib. Inhibitors of KIT, a membrane receptor, are mainly represented by imatinib, an inhibitor of tyrosine kinase. Finally, mTOR inhibitors act on the signaling pathway PI3K/AKT/mTOR, and key molecules are temsirolimus, everolimus, and deforolimus. PMID- 19299049 TI - [Successful treatment of life-threatening Panton-Valentine leucocidin positive Staphylococcus aureus pneumonia with antibiotics and immunoglobulins targeting the toxin production]. AB - We report a patient who presented a severe Panton-Valentine-secreting methicillin susceptible Staphylococcus aureus pneumonia with threatening multi-organ failure including acute respiratory distress syndrome, cardiac failure, renal failure and disseminated intravascular coagulation. Clinical and biological disease course using empiric therapy with treatment directed against toxin production (linezolid, clindamycin and intravenous immunoglobulins) was found to be quickly effective. PMID- 19299050 TI - [A rash. Seborrheic keratosis]. PMID- 19299051 TI - [A sciatica occurring in Behcet's disease]. PMID- 19299052 TI - Radiological monitoring: terrestrial natural radionuclides in Kinta District, Perak, Malaysia. AB - Natural background gamma radiation and radioactivity concentrations were investigated from 2003 to 2005 in Kinta District, Perak, Malaysia. Sample locations were distant from any 'amang' processing plants. The external gamma dose rates ranged from 39 to 1039 nGy h(-1). The mean external gamma dose rate was 222+/-191 nG yh(-1). Small areas of relatively enhanced activity were located having external gamma dose rates of up to 1039+/-104 nGy h(-1). The activity concentrations of (238)U, (232)Th and (40)K were analyzed by using a high resolution co-axial HPGe detector system. The activity concentration ranges were 12-426 Bq kg(-1) for (238)U, 19-1377 Bq kg(-1) for (232)Th and <19-2204 Bq kg(-1) for (40)K. Based on the radioactivity levels determined, the gamma-absorbed dose rates in air at 1m above the ground were calculated. The calculated dose rates and measured dose rates had a good correlation coefficient, R of 0.94. To evaluate the radiological hazard of the natural radioactivity, the radium equivalent activity, the gamma-absorbed dose rate and the mean population weighted dose rate were calculated. An isodose map for the Kinta District was also produced. PMID- 19299053 TI - Assessment of atmospheric sulfur with the epilithic moss Haplocladium microphyllum: evidences from tissue sulfur and delta34S analysis. AB - The application of geochemical signals in mosses is more and more popular to investigate the deposition of atmospheric pollutants, but it is unclear whether records of atmospheric sulfur in mosses differ between their diverse habitats. This study aimed to investigate the influence of growing condition on tissue sulfur and delta34S of Haplocladium microphyllum. Epilithic and terricolous mosses in open fields, mosses under different canopy conditions were considered. We found that tissue sulfur and delta34S of mosses under different habitats were not consistent and could not be compared for atmospheric sulfur research with each other even collected at the same site, moss sulfur and delta34S records would be distorted by subsoil and upper canopies in different degrees, which possibly mislead the interpretation of atmospheric sulfur level and sources. Consequently, mosses on open rocks can be used reliably to assess atmospheric derived sulfur in view of their identical sulfur and delta34S evidences. PMID- 19299054 TI - Pulsed gas injection: a minimum effort approach for enhanced natural attenuation of chlorobenzene in contaminated groundwater. AB - Chlorobenzene-contaminated groundwater was used to assess pulsed gas sparging as a minimum effort aeration strategy to enhance intrinsic natural attenuation. In contrast to existing biosparging operations, oxygen was supplied at minimum rate by reducing the gas injection frequency to 0.33 day(-1). Field tests in a model aquifer were conducted in a 12 m long reactor, filled with indigenous aquifer material and continuously recharged with polluted groundwater over 3 years. The closed arrangement allowed yield balances, cost accounting as well as the investigation of spatial distributions of parameters which are sensitive to the biodegradation process. Depending on the injection frequency and on the gas chosen for injection (pure oxygen or air) oxygen-deficient conditions prevailed in the aquifer. Despite the limiting availability of dissolved oxygen in the groundwater, chlorobenzene degradation under oxygen-deficient conditions proved to be more effective than under conditions with dissolved oxygen being available in high concentrations. PMID- 19299055 TI - Particle-size-fractioned transfer of dioxins from sediments to water columns by resuspension process. AB - Particle-size-fractioned transfer of dioxins from sediments to water columns by resuspension process was investigated, using supernatant samples obtained from shaking experiments of sediment-water pairs simulating natural disturbances. The concentrations (dry-matter mass basis) of individual compounds (C(fraction)) in two particle size fractions (0.1-1 and 1-10 microm) in the supernatants were generally slightly higher than those in the original sediment (C(sed)). C(fraction)/C(sed) ratios ranged from 0.45 to 5.9 (median 1.5) without consistent differences among congener groups or consistent correlations against the number of chlorine atoms. The dioxin concentrations in the water column associated with the remaining sediment particles can therefore be estimated by those in the original sediment and by the concentration of suspended sediment particles in the water. The concentration of each compound in the remaining sediment particles (mostly 0.1-10 microm in size) can be roughly estimated by multiplying the concentration in the original sediment by 1.5. PMID- 19299056 TI - Illicit drugs and pharmaceuticals in the environment--forensic applications of environmental data, Part 2: Pharmaceuticals as chemical markers of faecal water contamination. AB - This manuscript is part two of a two-part study aiming to provide a better understanding and application of environmental data not only for environmental aims but also to meet forensic objectives. In this paper pharmaceuticals were investigated as potential chemical indicators of water contamination with sewage. The monitoring program carried out in Wales revealed that some pharmaceuticals are particularly persistent and/or ubiquitous in contaminated river water and therefore might be considered as potential conservative or labile wastewater indicators. In particular, these include some anti-inflammatory/analgesics, antiepileptics, beta-blockers, some H2-receptor antagonists and antibacterial drugs. PMID- 19299057 TI - Chemical and ecotoxicological analyses of sediments and elutriates of contaminated rivers due to e-waste recycling activities using a diverse battery of bioassays. AB - A multi-trophic, multi-exposure phase assessment approach was applied to characterize the toxicity of sediments collected from two rivers in Guiyu, China, an e-waste recycling centre. Elutriate toxicity tests (bacterium Vibrio fischeri and microalga Selenastrum capricornutum) and whole sediment toxicity test (crustacean Heterocypris incongruens) showed that most sediments exhibited acute toxicity, due to elevated heavy metals and PAHs levels, and low pH caused by uncontrolled acid discharge. The survival rates of crustaceans were negatively (p < 0.05) correlated with total PAHs in sediments (411-1755 mg kg(-1)); EC50s of V. fischeri on the elutriates were significantly correlated with elutriate pH (p < 0.01). Significant (p < 0.05) correlations between the induction of hepatic metallothionein in tilapia (Oreochromis mossambicus) and metal concentrations (Cu, Zn, Pb) in sediments were also observed, when fish were fed with diets containing sediment. The results showed that uncontrolled e-waste recycling activities may bring adverse effects to local aquatic ecosystem. PMID- 19299058 TI - Problem-solving therapy for depression: a meta-analysis. AB - Problem-Solving Therapy (PST) is a cognitive-behavioral intervention that focuses on training in adaptive problem-solving attitudes and skills. The purpose of this paper was to conduct a meta-analysis of controlled outcome studies on efficacy of PST for reducing depressive symptomatology. Based on results involving 21 independent samples, PST was found to be equally effective as other psychosocial therapies and medication treatments and significantly more effective than no treatment and support/attention control groups. Moreover, component analyses indicated that PST is more effective when the treatment program includes (a) training in a positive problem orientation (vs. problem-solving skills only), (b) training in all four major problem-solving skills (i.e., problem definition and formulation, generation of alternatives, decision making, and solution implementation and verification), and (c) training in the complete PST package (problem orientation plus the four problem-solving skills). PMID- 19299059 TI - 'You are not yourself': exploring masculinities among heterosexual African men living with HIV in London. AB - It is now clear that gender is an essential factor shaping the narratives of men as well as women. However, there have been few studies of the daily lives or sexual activities of heterosexual men. Hence, strategies developed to prevent the spread of the HIV virus are rarely based on detailed knowledge of the men whose behaviours they are intended to change; this is especially evident in the developing world where the epidemic is most severe. Nor do we know very much about those men who have already been diagnosed as HIV positive. Around 13 million men are now living with HIV of whom around 96% are in low or middle income countries. Migrants from developing countries also make up the majority of positive people in a number of developed countries. In the UK, for example, heterosexual activity is now responsible for about half of all new HIV diagnoses with the majority of those involved being of African origin. But almost nothing is known about the ways in which different constructions of masculinity affect their experiences of illness. This study used qualitative methods to explore the experiences of a sample of black African men who defined themselves as heterosexual and were receiving treatment for HIV and/or AIDS in London. It explored their feelings, their needs, their hopes and their desires as they negotiated their lives in the diaspora. PMID- 19299060 TI - From pleasure to pain: The role of the MPQ in the language of phantom limb pain. AB - In opposition to the argument that pain is private, personal and unsharable, I propose that the intersubjectivity of pain is fundamental to it. Using the case of phantom limb, I show how a specific language of pain emerged and became concretized in the US circa 1975 with the advent of the McGill Pain Questionnaire (MPQ). Through widespread use of the MPQ, a language of pain materialized, one that was commonly used to describe the qualitative dimensions of phantom limb. After 1975, the terminology used within the medical literature was overwhelmingly consonant with the set of descriptors advanced by the MPQ. The utilization of a pain questionnaire to assess the qualitative dimensions of phantom limb effectively accentuated pain, and by 1980, what was once considered relatively rare became a common sequela of phantom manifestation. PMID- 19299061 TI - Effect of job strain and depressive symptoms upon returning to work after acute coronary syndrome. AB - The purpose of this study was 2-fold: to assess whether job strain and depressive symptoms were independent predictors of the timing of return to work after acute coronary syndrome, and to determine whether the association of job strain and timing of return to work was moderated by depressive symptoms. In this 6-month longitudinal study, a total of 240 employed women and men who were admitted to hospital with acute coronary syndrome were consecutively recruited in the United States and Japan. A structured interview and medical record review were conducted to collect baseline data during hospitalization. At 2 and 6months post hospitalization, we mailed a questionnaire to assess the timing of first day of return to work and presence of depressive symptoms. It was found that job strain was a significant independent predictor of a later return to work, even after controlling for potential confounding variables. In addition, when depressive symptoms were treated as a time-dependent covariate, patients who had Beck Depression Inventory-II scores of > or =6 to <16 or > or =16 were slower to return to work than patients with scores <6. An earlier return to work might be promoted by interventions focused on reducing psychological job demand, increasing perceived job control, and treating depressive symptoms. PMID- 19299062 TI - Walking on a line: a motor paradigm using rotation and reflection symmetry to study mental body transformations. AB - Researchers have recently reintroduced the own-body in the center of the social interaction theory. From the discovery of the mirror neurons in the ventral premotor cortex of the monkey's brain, a human embodied model of interindividual relationship based on simulation processes has been advanced, according to which we tend to embody spontaneously the other individuals' behavior when interacting. Although the neurocognitive mechanisms of the embodiment process have started being described, the mechanisms of self-location during embodiment are still less known. Here, we designed a motor paradigm which allows investigating in ecologically more valid conditions whether we embody another person's intransitive action with an embodied or disembodied self-location. Accordingly, we propose a phenomenological model of self-other interaction showing how perspective-taking mechanisms may relate on mental body transformation and offering a promising way to investigate the different sorts of intersubjectivity. PMID- 19299063 TI - Cardiac arrest after air travel successfully treated by presumptive fibrinolysis. AB - Thromboembolic events should be strongly suspected in cases where medical events occur after prolonged air travel. Pulmonary embolism should be suspected in patients with sudden cardiac arrest. Consequently, patients should be managed according to this eventuality. We report a case of sudden cardiac arrest after air travel in which the patient survived after fibrinolysis was prescribed on a presumptive basis. PMID- 19299064 TI - Integration of endocrine and mechanical signals in the regulation of myometrial functions during pregnancy and labour. AB - In this review, we describe a new model to explain the regulation of myometrial function during pregnancy and labour. We propose that the myometrium undergoes dramatic changes in phenotype from early pregnancy until the onset of labour, characterized by an early proliferative phase, an intermediate phase of cellular hypertrophy and matrix elaboration, a third phase in which the cells assume a contractile phenotype and the final phase in which cells become highly active and committed to labour. The last phase of myometrial differentiation is postpartum uterine involution, completing the reproductive cycle following pregnancy and labour by returning the uterus to its non-pregnant receptive state. We further propose that phenotypic modulation of the uterine myocytes is the result of integration of endocrine signals and mechanical stimulation of the uterus by the growing fetus. Our previous studies have shown that these signals are important in regulating the onset of labour and we now have indications that they regulate earlier myometrial smooth muscle differentiation. We show that the high rate of myometrial cell proliferation in early pregnancy which reflects important aspects of many smooth muscle populations during development. The proliferative phenotype was associated with dramatic changes in the expression of IGF family proteins and coincided with an up-regulation of the anti-apoptotic pathway. Preliminary evidence suggests that myometrial hyperplasia was controlled by the PI3K-Akt-mTOR signaling pathway. The modulation of the mTOR pathway by rapamycin blocked the proliferative activity of the uterine myocytes. The growth and remodeling of the myometrium during pregnancy was associated with increased synthesis of extra cellular matrix (ECM) proteins and their corresponding integrin receptors. Our results show a decrease in expression of fibrillar collagens and a coordinated temporal increase in expression of components of the basement membrane near term associated with decreased progesterone levels and increased mechanical tension. The phenotypic modulation of uterine smooth muscle cells during pregnancy culminates at term when a myometrium-specific conversion commits these cells to the labour phenotype, characterized by increased excitability, spontaneous activity, responsiveness to agonists and effective coupling of the myocytes. The reinforcement of the ECM-integrin interaction contributes to myometrial hypertrophy and remodeling during late pregnancy and facilitates force transduction during the contractions of labour by anchoring hypertrophied SMCs to the uterine ECM. In addition, we suggest that myometrial cells play an important role in the generation and regulation of uterine inflammation, which is a characteristic feature of parturition. We provide here substantial evidence that myometrial cells can actively participate in the inflammatory process in the uterus by the release of the pro-inflammatory chemokine MCP-1. The increased production of MCP-1 in the term myometrium was associated with uterine occupancy and regulated by progesterone, suggesting that mechanical and endocrine signals integrate to regulate the expression of the chemokine and the induction of labour. A better understanding of the mechanisms regulating myometrial differentiation during pregnancy might inform the development of new therapeutic strategies for the management of preterm labour, which remains a leading cause of neonatal morbidity and mortality. Our data are obtained mostly from the rat, but we believe that they are generally applicable across species. PMID- 19299065 TI - Avian sensitivity to mortality: prioritising migratory bird species for assessment at proposed wind farms. AB - Wind power generation is likely to constitute one of the most extensive human physical exploitation activities of European marine areas in the near future. The many millions of migrating birds that pass these man-made obstacles are protected by international obligations and the subject of public concerns. Yet some bird species are more sensitive to bird-wind turbine mortality than others. This study developed a simple and logical framework for ranking bird species with regard to their relative sensitivity to bird-wind turbine-collisions, and applied it to a data set comprising 38 avian migrant species at the Nysted offshore wind farm in Denmark. Two indicators were selected to characterize the sensitivity of each individual species: 1) relative abundance and 2) demographic sensitivity (elasticity of population growth rate to changes in adult survival). In the case study from the Nysted offshore wind farm, birds of prey and waterbirds dominated the group of high priority species and only passerines showed a low risk of being impacted by the wind farm. Even where passerines might be present in very high numbers, they often represent insignificant segments of huge reference populations that, from a demographic point of view, are relatively insensitive to wind farm-related adult mortality. It will always be important to focus attention and direct the resources towards the most sensitive species to ensure cost effective environmental assessments in the future, and in general, this novel index seems capable of identifying the species that are at high risk of being adversely affected by wind farms. PMID- 19299066 TI - Combustion in porous media and its applications--a comprehensive survey. AB - The rapid advances in technology and improved living standard of the society necessitate abundant use of fossil fuels which poses two major challenges to any nation. One is fast depletion of fossil fuel resources; the other is environmental pollution. The porous medium combustion (PMC) has proved to be one of the technically and economically feasible options to tackle the aforesaid problems to a remarkable extent. PMC has interesting advantages compared with free flame combustion due to the higher burning rates, the increased power dynamic range, the extension of the lean flammability limits, and the low emissions of pollutants. This article provides a comprehensive picture of the global scenario of research and developments in PMC and its applications that enable a researcher to decide the direction of further investigation. The works published so far in this area are reviewed, classified according to their objectives and presented in an organized manner with general conclusions. A separate section is devoted for the numerical modeling of PMC. PMID- 19299067 TI - Cesium-137 monitoring using mosses from W. Macedonia, N. Greece. AB - (137)Cs activities in mosses and substrate (soil, bark) collected from W. Macedonia, Greece were measured 20 years after the Chernobyl reactor accident. Archive material from previous studies was also used for comparison and diachronic estimation of the radio-contamination status. A gradual decrease was detected which depended on various factors such as the collected species, location, growth rate and substrate. Maximum accumulation capacity of (137)Cs was observed in the epilithic mosses in comparison to the epiphytic ones. The (137)Cs content in the bark of the two broad-leaved species (oak and fagus) was higher than that of the conifer (pinus). Bark specimens of about 50 cm height were in general more contaminated than those of 200 cm. Autoradiography revealed an amount of (137)Cs distributed more or less uniformly in moss thalli. The high (137)Cs activities found in mosses 20 years after Chernobyl suggest that these primitive plants are effective, suitable and inexpensive biological detectors of the distribution and burden of radionuclide fallout pattern. PMID- 19299068 TI - Editorial comment on: Laparoscopic radical nephroureterectomy: a multicenter analysis in Japan. PMID- 19299069 TI - Tumour grade, treatment, and relative survival in a population-based cohort of men with potentially curable prostate cancer. AB - BACKGROUND: There is insufficient information regarding the benefit of treatment with curative intent for men with localised poorly differentiated prostate cancer (PCa). OBJECTIVE: To evaluate relative survival in men with potentially curable PCa in relation to Gleason score (GS) and treatment as practiced in the community at large. DESIGN, SETTING, AND PARTICIPANTS: A population-based study including all men with localised PCa registered in Sweden's National Prostate Cancer Register. INTERVENTIONS: Hormonal therapy, watchful waiting, and treatment with curative intent. MEASUREMENTS: The ratio of observed deaths to expected deaths, determined from survival in the general male population of the same age, was assessed using Poisson regression analysis, with GS and treatment as covariates. Interaction between GS and treatment was tested in a multivariate Cox proportional hazard analysis. RESULTS AND LIMITATIONS: A total of 31,903 men with potentially curable tumour (T1-T3, N0/NX, M0/MX, age <75 yr, and prostate specific antigen [PSA] <20 ng/ml) were identified. GS was recorded for 28,454 of these men. Some 19,606 men (60.8%) were treated with curative intent, and 12,645 men (39.2%) were given either hormonal treatment or expectant management. The ratios between observed and expected survival gradually increased for men with GS 10, with GS to 3.3 for men treated conservatively and to 1.4 for men treated with curative intent. There was a significant interaction between GS and treatment, with a relatively greater benefit from treatment with curative intent for men with high-grade tumours. The results have to be interpreted with some caution, as there was no randomisation between the treatment groups. CONCLUSIONS: Survival for men with well-differentiated tumours is close to that of the general population, regardless of treatment, but the outcome is dismal for men with poorly differentiated tumours, whichever treatment is applied. Nevertheless, men with poorly differentiated tumours benefit more from curative treatment than do men with well- differentiated tumours. PMID- 19299070 TI - Editorial comment on: Laparoscopic radical nephroureterectomy: a multicenter analysis in Japan. PMID- 19299071 TI - Editorial comment on: tumor grade, treatment, and relative survival in a population-based cohort of men with potentially curable prostate cancer. PMID- 19299072 TI - Laparoscopic radical nephroureterectomy: a multicenter analysis in Japan. AB - BACKGROUND: Laparoscopic nephroureterectomy (LNUx) is prevalent in Japan and throughout the world, but long-term outcome data remain limited. OBJECTIVE: To understand the present state of LNUx in Japan, we conducted a multicenter analysis of clinical outcome and long-term cancer control for patients who underwent the procedure. DESIGN, SETTING, AND PARTICIPANTS: Between January 1995 and December 2005, 1003 patients with urothelial cancer in the upper urinary tract were treated with LNUx at 51 institutions in Japan, and patient data were collected retrospectively. MEASUREMENTS: Patient profiles were gathered and analyzed for survival, intravesical recurrence, and risk factors influencing them. RESULTS AND LIMITATIONS: Median operative time was 320 min. Median bleeding volume was 232 ml. Complications occurred in 93 cases (9.3%) intraoperatively and in 107 cases (10.7%) postoperatively. Overall survival rate was 70% at 5 yr. Grade 3, pT3 or pT4, multifocal tumor, lymph-node metastasis, and previous or coexistent bladder tumor were independent risk factors for overall survival. Intravesical recurrence rate was 43% at 5 yr. Intravesical recurrence occurred more frequently in males, in patients with multifocal tumors, in patients with previous or coexistent bladder tumors, and in patients who underwent the hand assisted approach. CONCLUSIONS: Our report represents the largest multicenter analysis of LNUx reported to date. Male sex and the use of the hand-assisted approach were shown for the first time to be risk factors for recurrence-free survival and intravesical recurrence. To further analyze the effectiveness of LNUx, a long-term outcome comparison with risk stratification must be made between LNUx and open nephroureterectomy. PMID- 19299073 TI - Editorial comment on: tumour grade, treatment, and relative survival in a population-based cohort of men with potentially curable prostate cancer. PMID- 19299074 TI - Editorial comment on: Laparoscopic radical nephroureterectomy: a multicenter analysis in Japan. PMID- 19299075 TI - Small interfering RNA targeting the subunit ATP6L of proton pump V-ATPase overcomes chemoresistance of breast cancer cells. AB - One of the mechanisms of multiple drug resistance (MDR) is inappropriate sequestration of basic chemotherapeutic agents in acidic endo-lysosomes of cells. The protonation, sequestration, and secretion (PSS) model indicates that drug distribution can be affected by intracellular pH such as lysosomal pH. The vacuolar-H(+)-ATPase (V-ATPase) plays an important role in regulation of intracellular pH by pumping protons into acidic endosomes via an ATP-driven process. In this study, ATP6L, the 16kDa subunit of V-ATPase, was knocked-down by anti-ATP6L small interfering RNA (siRNA) to study the effect on chemosensitivity in the human drug-resistant breast cancer cells MCF-7/ADR. Introduction of anti ATP6L small interfering RNA duplex into drug-resistant cancer cells significantly inhibited the expression of ATP6L mRNA and protein, as detected by qRT-PCR and Western blot. Inhibition of ATP6L expression by siRNA in MCF-7/ADR sensitized the cells to the cytotoxicity of basic chemotherapeutic agents like doxorobicin, 5 fluorourocil and vincristine. This effect was mediated by a significant increase in lysosomal pH and retention of anticancer drugs into nuclei of cells. These results support the role of tumor acidity in resistance to chemotherapy and provide a rationale for the use of tumor pH modifier agents as coadjuvants in novel anticancer therapies. PMID- 19299076 TI - Identification of metastasis-associated proteins involved in gallbladder carcinoma metastasis by proteomic analysis and functional exploration of chloride intracellular channel 1. AB - Advanced gallbladder cancer has an extremely poor prognosis because of metastasis. Identification of metastasis-related biomarkers is essential to improve patient survival. In the present study, metastasis-associated proteins were identified by comparative proteomic analysis and the metastasis-related function of the candidate protein, chloride intracellular channel 1 (CLIC1), was further elucidated. Two cell lines with high or low metastatic potential (termed GBC-SD18H and GBC-SD18L, respectively), originating from the same parental gallbladder carcinoma GBC-SD cell line, were identified by spontaneous metastasis in vivo and characterized by metastatic phenotypes analysis in vitro. Subsequently, a proteomic approach comprised of two-dimensional gel electrophoresis analysis and mass spectroscopy was used to identify and compare the protein expression patterns between GBC-SD18L and GBC-SD18H. Twenty-six proteins were identified and further verified by one-dimensional Western blotting and semiquantitative reverse transcriptase polymerase chain reaction analysis. It was determined that CLIC1, ezrin, vimentin, annexin A3, WD repeat domain 1, triosephosphate isomerase, C1-tetrahydrofolate synthase, Rho GDP-dissociation inhibitor 1, T-complex protein 1, heterogeneous nuclear ribonucleoprotein K, glutamate dehydrogenase 1, proteasome activator complex subunit 3 and Rab GDP dissociation inhibitor beta were significantly up-regulated in the highly metastatic GBC-SD18H cell line compared to the poorly metastatic GBC-SD18L cell line. However, phosphoglycerate kinase 1 and programmed cell death protein 8 were significantly down-regulated in the highly metastatic GBC-SD18H cell line compared to GBC-SD18L. Considering that CLIC1 was profuse in highly metastatic GBC-SD18H but scarce in poorly metastatic GBC-SD18L, the association of CLIC1 with metastasis was further elucidated by the overexpression and RNA interference of CLIC1 in GBC-SD18L cells and GBC-SD18H cells, respectively. The results demonstrated that the overexpression of CLIC1 promoted cell motility and invasion of GBC-SD18L in vitro, while RNA interference of CLIC1 remarkably decreased cell motility and invasive potency of GBC-SD18H in vitro, indicating that CLIC1 might play an important role in metastasis of gallbladder carcinoma. PMID- 19299077 TI - Anti-tumor activity of a novel EGFR tyrosine kinase inhibitor against human NSCLC in vitro and in vivo. AB - Epidermal growth factor receptor (EGFR) is highly expressed in many human tumors including non-small cell lung cancer (NSCLC). Treatment with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) has led to dramatic clinical improvement in selected patients with NSCLC. However, intrinsic and acquired resistance to EGFR-TKI remains a common phenomenon. Novel EGFR-TKI, structurally different with erlotinib or gefitinib might be beneficial for patients with NSCLC. In this study, we examined the anti-tumor effect of a newly synthesized novel EGFR tyrosine kinase inhibitor N-(3-chloro-4-fluorophenyl)-N-(7 methoxy-6-(3-morpholinopropoxy) quinazolin-4-yl)-3,3-dimethylbutanamide (F90). In vitro studies in a panel of three different human NSCLC cell lines revealed that F90 inhibited cell proliferation with high potency and induced G0/G1 arrest of cell cycle and apoptosis. F90 markedly reduced phosphorylation of EGFR and inhibited activation of MAPK and Akt. Oral administration of F90 (80mg/kg/day) to BALB/c nude mice bearing NSCLC cell lines xenografts significantly retarded tumor growth. In conclusion, F90 has potent anti-tumor activity on human lung cancer in vitro and in vivo. PMID- 19299078 TI - TRAIL as a target in anti-cancer therapy. AB - The tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily that can initiate apoptosis through the activation of their death receptors. The ability of TRAIL to selectively induce apoptosis of transformed or tumor cells but not normal cells promotes the development of TRAIL based cancer therapy. Accumulating preclinical studies demonstrate that the TRAIL ligand can effectively induce cancer cell apoptosis. Completed and ongoing Phases I and II clinical trials using TRAIL are showing clinically promising outcomes without significant toxicity. Importantly, TRAIL, DR4 and DR5 can all be induced by chemotherapeutics and/or radiation, which can sensitize cancer cells to TRAIL. Thus, understanding the regulation of the TRAIL apoptosis pathway can help develop more selective TRAIL-based agents for the treatment of human cancer. PMID- 19299079 TI - Epigenetic dysregulation of secreted Frizzled-related proteins in multiple myeloma. AB - We analysed the clinical impact of epigenetic dysregulation of the Wnt pathway in malignant plasma cell disorders. In multiple myeloma (MM) cell lines, aberrant promoter hypermethylation of the secreted Frizzled-related protein (SFRP) genes was a common event, and hypermethylation of SFRP1,-2 and -5 was associated with transcriptional silencing. Among 76 primary patient samples, the frequency of aberrant methylation was 35.5% for SFRP1, 52.6% for SFRP2, 1.3% for SFRP4 and 6.9% for SFRP5. Hypermethylation of SFRP1 and -2 genes was detected in monoclonal gammopathy of undetermined significance and all MM stages including plasma cell leukaemia (PCL), while SFRP5 methylation was restricted to advanced MM stages and PCL. Our data indicate that epigenetic silencing of Wnt antagonists is an early event in MM pathogenesis and that SFRP5 hypermethylation may play a role in disease progression. PMID- 19299080 TI - Utilization of industrial waste products as pozzolanic material in cemented paste backfill of high sulphide mill tailings. AB - In this study, the potential use of the industrial waste products including waste glass (WG), fly ash (FA), granulated blast furnace slag (GBFS) and silica fume (SF) as pozzolanic additive for the partial replacement of ordinary Portland cement (OPC) in cemented paste backfill (CPB) of sulphide-rich mill tailings was investigated. The influence of these industrial waste products on the short- and long-term mechanical performance of CPB was demonstrated. The rate of development of strength of CPB samples tended to slow down when the pozzolanic wastes were incorporated or increased in dosage in the binder phase. Severe losses (by 26%) in the strength of CPB samples produced from exclusively OPC occurred after an initial curing period of 56 days. The addition of WG (10-30 wt%) as a partial replacement of OPC was observed to aggravate further the strength losses of CPB samples. GBFS, FA and SF appeared to improve the long-term performance of CPB samples; albeit, only GBFS and SF could be incorporated into the binder phase only at certain levels i.e. up to 20 wt% GBFS and 15wt% SF in order to maintain a threshold strength level of 0.7MPa over 360 days. SEM studies have provided further insight into the microstucture of CPB and confirmed the formation of deleterious gypsum as the expansive phase. These findings have demonstrated that the industrial waste products including GBFS and SF can be suitably used as mineral additives to improve the long-term mechanical performance of CPB produced from sulphide-rich tailings as well as to reduce the binder costs in a CPB plant. PMID- 19299081 TI - Determination of selenium in natural waters by adsorptive differential pulse cathodic stripping voltammetry. AB - In this work bovine albumin was used innovatively as a medium for adsorptive accumulation of Se-I(2) on thin mercury film electrode. Se-I(2) was formed by reaction between Se(IV) and iodide in HCl media. The adsorbed Se-I(2) was stripped in 0.05 M HCl by differential pulse cathodic potential scan. The proposed method was successfully applied to analysis of Se(IV) and Se(VI) in natural waters sampled from some lagoons south of Caspian Sea. The optimum reaction conditions and other analytical parameters and influence of cations and anions were studied. The detection limit was 0.37 ng mL(-1). The obtained results were compared with the results of DPCSV after electrochemical preconcentration, HG-AAS and ICP-AES. PMID- 19299082 TI - A sequential extraction to determine the distribution of phosphorus in the seawater and marine surface sediment. AB - The distribution of phosphorus species among environmental compartments (e.g., between water and sediment), significantly affects the bioavailability of these species to organisms. The eastern Mediterranean Sea is one of the most extreme oligotrophic oceanic regions on earth in terms of nutrient concentrations and primary productivity. The paper presents the results of inorganic and organic forms in surface sediment and seawater from NE Mediterranean Sea (Burclar Bay, Erdemli, South Anatolia of Turkey) in May 2007. Speciation of phosphorus in seawater and surface sediment using inductively coupled plasma-atomic emission spectrometer (ICP-AES) has been reported here. The method is based on sequential extractions of the seawater and sediment each releasing four forms of inorganic phosphorus: loosely sorbed phosphorus, phosphorus bound to aluminium (P-Al), phosphorus bound to iron (P-Fe) and phosphorus bound to calcium (P-Ca). The most abundant form of inorganic phosphorus in the seawater and surface sediment is calcium-bound phosphorus. Relative abundances of the remaining phosphorus forms in sediment follow the order: P-Al>P-Fe>loosely bound-P and in seawater follow the order P-Fe>P-Al>loosely bound-P. PMID- 19299083 TI - Removal of lead and zinc ions from water by low cost adsorbents. AB - In this study, activated carbon, kaolin, bentonite, blast furnace slag and fly ash were used as adsorbent with a particle size between 100 mesh and 200 mesh to remove the lead and zinc ions from water. The concentration of the solutions prepared was in the range of 50-100 mg/L for lead and zinc for single and binary systems which are diluted as required for batch experiments. The effect of contact time, pH and adsorbent dosage on removal of lead and zinc by adsorption was investigated. The equilibrium time was found to be 30 min for activated carbon and 3h for kaolin, bentonite, blast furnace slag and fly ash. The most effective pH value for lead and zinc removal was 6 for activated carbon. pH value did not effect lead and zinc removal significantly for other adsorbents. Adsorbent doses were varied from 5 g/L to 20 g/L for both lead and zinc solutions. An increase in adsorbent doses increases the percent removal of lead and zinc. A series of isotherm studies was undertaken and the data evaluated for compliance was found to match with the Langmuir and Freundlich isotherm models. To investigate the adsorption mechanism, the kinetic models were tested, and it follows second order kinetics. Kinetic studies reveals that blast furnace slag was not effective for lead and zinc removal. The bentonite and fly ash were effective for lead and zinc removal. PMID- 19299084 TI - Modelling disinfection by-products formation in bromide-containing waters. AB - A kinetic model capable of simulating by-products formation in bromide-containing waters during disinfection processes is presented in this paper. The model is based on two parallel sequences of incorporation and oxidation reactions induced by bromine or chlorine reacting with natural organic matter (NOM). Each sequence starts from a different type of NOM functionality that has its own set of specific reaction rate. Decay reactions of NOM and halogenated intermediates are assumed to follow a first order kinetic, while disinfection by-product (DBP) generation reactions are simulated introducing so-called splitting coefficients. This approach allows obtaining explicit expressions for DBP species. Model's results are compared with experimental data obtained for seawater samples. Comparison of the data confirms the model's ability to predict DBPs formation with high precision. PMID- 19299085 TI - Factors affecting simultaneous nitrification and de-nitrification (SND) and its kinetics model in membrane bioreactor. AB - Experiments have been carried out to investigate the effect of biological factors such as dissolved oxygen (DO), food/microorganism (F/M) ratio, carbon/nitrogen (C/N) ratio and pH on performance of SND in membrane bioreactor (MBR). It was found that a low DO was advantageous to SND on condition that nitrification was not inhibited, while F/M ratio and C/N ratio have reverse effects on SND, and pH should also be controlled in a suitable range. Based on the conventional activated sludge model, a deduction was conducted to illustrate that SND could take place from the theoretical aspect, and it was proved that high organics was effective in improving SND. In addition, a kinetic model for SND was constituted on the basis of batch test result, and the simulation nitrate saturation coefficient K(NO)(3) was much higher than that in a single-sludge wastewater treatment system. PMID- 19299086 TI - Study on the catalytic effect of NiO nanoparticles on the thermal decomposition of TEGDN/NC propellant. AB - The catalytic effect of NiO nanoparticles on the thermal decomposition of double base propellant composed of nitrocellulose (NC) and triethylene glycol dinitrate (TEGDN) has been investigated by thermogravimetry-mass spectrometry (TG-MS) coupling technique. It was shown that adding 2% of NiO nanoparticles to TEGDN/NC propellant can accelerate the thermal decomposition process after around 188 degrees C TG-MS analysis indicated that NiO nanoparticles have resulted in the increase in intensity (peak area) of m/z=27, 28, and 29 MS signals, but the decrease in the intensity of m/z=18, 30, 44 and 46 (peak 2) MS signals during the thermal decompsition. The catalytic mechanism was also discussed in this paper. PMID- 19299087 TI - The long and the short of treatments for alcohol or cannabis misuse among people with severe mental disorders. AB - Substance misuse among people with severe mental disorders is very common and has been associated with a number of adverse outcomes. Most trials among people with severe mental disorders have reported general substance misuse outcomes and have argued that longer treatments are likely to be more effective. Such studies have not specifically compared intervention effectiveness for alcohol vs. cannabis misuse. The present paper reports findings from a synthesis of datasets from three randomized controlled trials (N=248) comparing brief (1-session advice or motivational interviewing, MI) and extended (10 sessions MI/cognitive-behaviour therapy; CBT) interventions for substance misuse among people with psychosis or major depression and outcomes are reported separately for alcohol and cannabis use. Whilst there were comparable rates of reduction in alcohol consumption for the BI and intensive MI/CBT, BI tended to be associated with less reduction in cannabis use. A stepped care approach for substance misuse in people with severe mental disorders may help to improve cannabis outcomes, including monitoring of both mental health and substance misuse domains, with interventions stepped up or down according to treatment response. Future studies should evaluate the effectiveness of stepped care approaches. PMID- 19299088 TI - Methodological issues in evaluating environmental risk factors on "apples and pears": a response to Logan. PMID- 19299089 TI - Comments on "Pelvic floor muscle exercise may improve female sexual function". PMID- 19299090 TI - Inhibition of Behcet's disease by calcitonin. AB - Behcet's disease (BD) is a chronic, multisystem inflammatory disorder characterized by relapsing oral aphthous and genital ulcers, ocular inflammation, erythemanodosum and folliculitis-like lesions of the skin, arthritis, and central nervous system involvement. Its pathogenesis has not been fully elucidated but the etiology is accepted to be multifactorial, therefore the treatment of Behcet's disease continues to be a major therapeutic challenge. The identification of novel therapeutic agents for the treatment of these disorders is important. Calcitonin (CT), a peptide hormone secreted in response to hypercalcemia, has the dual effect of inhibiting osteoclast recruitment as well as their resorptive activity. A number of reviews have concluded that salmon calcitonin is safe and effective in the treatment of osteoporosis. Calcitonin abrogated the stimulating effect of RANKL or prednisolone; similar results were obtained with OPG. Additionally, the analgesic activity of salmon calcitonin has been shown in several controlled prospective double-blind studies to improve pain. Exogenous calcitonin is thought to cross the blood-brain barrier and to accumulate slowly in the brain, inducing analgesia once sufficient receptors are occupied. Since CT could antagonize resorptive and analgesic activity by competitively binding to CTR and has been considered as a specific antagonist, we postulate that the CT could function as a novel agent to inhibit BD. In our opinion, if the hypothesis proved to be practical, CT could be widely used in clinical settings to treat BD. PMID- 19299091 TI - [Genetic recombination in vaccine poliovirus: comparative study in strains excreted in course of vaccination by oral poliovirus vaccine and circulating strains]. AB - AIM OF STUDY: Recombination is one of the major mechanisms of evolution in poliovirus. In this work, recombination was assessed in children during vaccination with OPV and among circulating vaccine strains isolated in Tunisia during the last 15 years in order to identify a possible role of recombination in the response to the vaccine or the acquisition of an increased transmissibility. MATERIAL AND METHODS: This study included 250 poliovirus isolates: 137 vaccine isolates, excreted by children during primary vaccination with OPV and 113 isolates obtained from acute flaccid paralytic (AFP) cases and healthy contacts. Recombination was first assessed using a double PCR-RFLP, and sequencing. RESULTS: Nineteen per cent of recombinant strains were identified: 20% of strains excreted by vaccinees among 18% of circulating strains. The proportion of recombinant in isolates of serotype1 was very low in the two groups while the proportions of recombinants in serotypes 2 and 3 were different. In vaccinees, the frequency of recombinants in serotype3 decreased during the course of vaccination: 54% after the first dose, 32% after the second and 14% after the third dose. CONCLUSION: These results suggest that recombination enhances the ability of serotype3 vaccine strains to induce an immune response. Apart from recent vaccination, it may contribute to a more effective transmissibility of vaccine strains among human population. PMID- 19299092 TI - [The latest TH17 lymphocyte in the family of T CD4+ lymphocytes]. AB - In recent years the T CD4+ lymphocyte family has grown. In the initial two components TH1 and TH2 lymphocytes were added the TH17 lymphocyte and T cell regulator (Treg). Under the influence of transforming growth factor beta, interleukin 6 (IL6), IL21 and IL23, the naive lymphocyte T CD4+ differentiates in TH17. Currently, the TH17 is recognized as the leading actor of local inflammation through the pro-inflammatory cytokines (interleukins 17, 21, 22) that secretes and the expansion and recruitment of neutrophils that leads. Therefore, it is involved in chronic inflammatory processes, autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosus), allergy and rejection of allogeneic transplants. TH17 lymphocyte opens up new therapeutic prospects for these pathologies. PMID- 19299093 TI - [Epigenome: the immersed part of the iceberg or the hidden face of pathologies]. PMID- 19299094 TI - Occurrence of non-sorbitol fermenting, verocytotoxin-lacking Escherichia coli O157 on cattle farms. AB - Escherichia coli O157 is often associated with hemorrhagic colitis and the hemolytic uremic syndrome (HUS). The verocytotoxins are considered to be the major virulence determinants. However, vt-negative E. coli O157 were recently isolated from patients with HUS. Several transmission routes to humans are described, but cattle feces are the primary source from which both the food supply and the environment become contaminated with E. coli O157. In a prevalence study performed on dairy, beef, mixed dairy/beef and veal farms in the summer of 2007, vt-negative isolates were detected on 11.8% (8/68) of the positive farms. From these eight farms, a total of 43 sorbitol-negative E. coli O157:H7 were collected. On five farms, only strains negative for the vt genes were present whereas both vt-negative and vt-positive strains could be detected on three other farms. Further characterization revealed that all isolates carried the eaeA and hlyA genes. Pulsed-field gel electrophoresis (PFGE) of all isolates resulted in nine different PFGE types and within the vt-negative strains, four different genotypes were identified, indicating that certain genetic clones are widespread over the cattle population. PMID- 19299095 TI - Morphological development and characterization of aromatase and estrogen receptors alpha and beta in fetal ovaries of cattle from days 110 to 250. AB - To better understand the role of estradiol-17beta in fetal ovarian development, presence and localization of cytochrome P450 aromatase (P450arom) and estrogen receptors alpha (ERalpha) and beta (ERbeta) proteins were characterized in fetal ovaries of cattle using immunohistochemistry. Fetal cattle ovaries were collected from an abattoir and sorted into fetal age groups (days 110, 130, 150, 170, 190, 210, 230, 250+) based on crown-rump length. In addition to immunohistochemistry, morphological analysis of ovarian and follicular formation was made. Ovaries appeared lobular at day 110, but by the end of gestation (day 250+) ovaries were oval-shaped similar to those found in adult animals. Ovarian structures within different lobes appeared to be at different developmental stages. At day 110, oocytes and pre-granulosa cells were observed in ovigerous cords that were still open to the surface epithelium. Most ovigerous cords appeared to be closed to the surface epithelium on day 130, all closed by day 150 and were no longer present at day 210. Ovarian follicles were classified as follows: Type 1(primordial): single layer of flattened granulosa cells, Type 1a (transitory): single layer of mixed flattened and cuboidal granulosa cells, Type 2 (primary): at least one but less than two layers of cuboidal granulosa cells, Type 3 (small preantral): two to three layers of granulosa cells, Type 4 (large preantral): four to six layers of granulosa, and the theca layer is forming around the follicle, Type 5 (antral): contain greater than six layers of granulosa cells, several layers of theca cells and the antrum has formed. Type 1 follicles were observed in day 110 ovaries. Follicle Types 1a and 2 were first observed on day 130. Type 3 follicles were first observed on day 150 and Types 4 and 5 were first observed on day 170. P450arom protein was localized in granulosa cells of follicle Types 2-5 and cells of rete tubules throughout the experimental period. There was punctate expression within stroma and rete masses. There was ERalpha protein localization in pre granulosa cells and germ cells of ovigerous cords and all surface epithelial cells. There was also localization in granulosa cells and oocytes of all follicle types and cells of rete tubules. There was punctate ERalpha protein expression in stroma and rete masses. ERbeta protein was localized in pre-granulosa cells and germ cells of ovigerous cords. Expression was also localized to granulosa cells of all follicle types and cells of rete tubules. ERbeta protein was punctate in oocytes of follicles, surface epithelial cells, stroma and rete masses. Thus, the fetal ovary of cattle has the steroidogenic enzyme (P450arom) to convert androgens to estradiol-17beta, and estrogen receptors alpha and beta to facilitate an estrogen response within the fetal ovary. PMID- 19299096 TI - [Male gonococcal infection: antibiotic resistance in the Monastir Teaching Hospital, Tunisia (1999-2003)]. PMID- 19299097 TI - A review of molecular imaging studies reaching the clinical stage. AB - The practice of molecular imaging in the clinics is examined across various imaging modalities to assess the current status of clinical molecular imaging. The various physiologic and scientific bases of clinical molecular imaging are surveyed to assess the possibilities and opportunities for the deployment of the different imaging modalities in the near future. The requisites for successful candidate(s) of clinical molecular imaging are reviewed for future development. PMID- 19299098 TI - Development of LC chiral methods for neutral pharmaceutical related compounds using reversed phase and normal phase liquid chromatography with different types of polysaccharide stationary phases. AB - The enantioselectivity of a collection of neutral pharmaceutical compounds on six different types of polysaccharide chiral stationary phases (CSPs), Chiralpak AD (and AD-RH), Chiralcel OD (and OD-RH), Chiralpak OJ (and OJ-R), Chiralcel AS (and AS-RH), Sepapak-2 and Sepapak-4 are compared using reversed phase (RPLC) and normal phase liquid chromatography (NPLC). Screening strategies for maximizing the probability of achieving an initial chiral separation hit for neutral compounds using both RPLC and NPLC are described. Further method optimizations are demonstrated by modifying parameters such as organic modifier composition, eluent pH or CSP particle size. Several practical examples of the application of chiral methods for the study of synthetic reaction mixtures are presented. The most critical validation aspects, including limit of detection, specificity, and ruggedness, are also briefly presented. PMID- 19299099 TI - A quantitative NMR protocol for the simultaneous analysis of atropine and obidoxime in parenteral injection devices. AB - A rapid selective and accurate quantitative (1)H NMR method was developed for the simultaneous analysis of obidoxime chloride and atropine sulfate, the active components in parenteral injection devices (PID) used for the emergency treatment of poisoning by toxic organophosphates. The spectra were acquired in 90% H(2)O 10% D(2)O using sodium 3-(trimethylsilyl)-1-propane sulfonate hydrate as the internal standard. Both synthetic mixtures and dosage forms were assayed. The results were compared with those obtained from a reported HPLC method. PMID- 19299100 TI - Surgical guidelines for the management of breast cancer. PMID- 19299101 TI - [Comparative evaluation of single-use gum-elastic bougie in a manikin-simulator (SimMAN)]. PMID- 19299102 TI - [Deleterious role of hyperthermia in neurocritical care]. AB - Fever is a secondary brain injury and may worsen neurological prognosis of neurological intensive care unit (NICU) patients. In response to an immunological threat, fever associates various physiological reactions, including hyperthermia. Its definition may vary but the most commonly used threshold is 37.5 degrees C. In animal studies, hyperthermia applied before, during or after cerebral ischemia may increase the volume of ischemic lesions. The mechanism of this effect may include increase in blood brain barrier permeability, increase in excitatory amino acid release and increase in free radical production. In NICU patients, fever is frequent, occurring in up to 20-30% of patients. Moreover, after haemorrhagic stroke, fever has been reported in 40-50% of patients. In half of the patients, fever may be related to an infectious cause but in more than 25% of patients, hyperthermia may be of central origin. After ischemic stroke, hyperthermia during the first 72 hours is associated with an increase in infarct size and increase in morbidity and mortality. This holds true also after subarachnoid haemorrhage. After traumatic brain injury, fever is not related to mortality but may increase morbidity. Whereas no causal link has been established between fever and unfavourable outcome, it seems reasonable to treat hyperthermia in patients suffering from brain injuries. In such patients, antipyretics have a moderate efficacy. In case of failure, they should be replaced by physical cooling techniques. PMID- 19299103 TI - [Incidence and evolution of thrombotic images within the internal jugular vein following Swan-Ganz catheter insertion in cardiac surgery]. AB - OBJECTIVES: Insertion of Swan-Ganz catheter for a few days may be necessary in cardiac surgery. This study was aimed at determining the incidence and the evolution of thrombotic images within the internal jugular vein as well as assessing their association with the presence of a prolonged fever at postoperative day 7 in the lack of any documented infection. MATERIAL AND METHODS: All the patients undergoing cardiac surgery had a two-dimensional ultrasonography of internal jugular veins preoperatively, at discharge (day 7) and at postoperative day 90 if thrombotic images were seen at day 7. RESULTS: Sleeve-like and compact thrombotic images have been observed in site of venipuncture in 52 patients (70.3%). None had any residual thrombotic image 90 days after the operation. No clinical thromboembolic migration has been observed. There was no statistical association between the presence of a thrombotic image at the ultrasonography and the duration of catheterization. Moreover, there was no association between the anticoagulation before, during and after the surgery and the presence of a thrombotic image. We found a non-significant association between fever at day 7 and the presence of a thrombotic image within the internal jugular vein. CONCLUSION: Thrombotic images in the internal jugular vein after catheterization are frequent and disappear at day 90. The limited sample size of this study does not provide strong evidence of the role of jugular thrombi in the prolongation of fever after cardiac surgery. PMID- 19299104 TI - [Ketamine for prevention of postoperative pain: what are the doses and indications?]. PMID- 19299105 TI - [Results of a national survey about the use of sedation scales in emergency prehospital medicine]. AB - The primary goal of sedation in emergency prehospital care is to guarantee the security of the mechanically ventilated patients by optimising their adaptation to the respirator. If the French prehospital guidelines are well codified, their applicability in routine clinical practice seem to be rather empirical. The aim of this national survey was to evaluate the use of the clinical sedation scales by the prehospital physicians. This prospective and clinical practice survey was begun in January 2005. An anonymous questionnaire was sent to the physicians working in the 377 Mobile Intensive Care Unit of the 105 French Emergency Medical Service System. The total response rate from physicians was 28% (n=497). Only 29% of the physicians (n=145) declared to use a sedation scale for a mechanically ventilated patient. The Ramsay score was used in 97% of the cases (n=141).The principal reasons given by the physicians for not using the sedation scales were their ignorance in 57% of the cases (n=200) and the systematic choice of a deep sedation in 42% of the cases (n=147). For 18% of them (n=62), the use of sedation scores was considered too complicated. The final results show that the utilisation ratio of the sedation scores is very low in emergency prehospital medicine and suggest that an effort toward improving the use of sedation in prehospital emergency medicine is necessary. PMID- 19299106 TI - Solid pseudopapillary tumor of the pancreas in a 14-year-old girl. PMID- 19299107 TI - [Primary hyperaldosteronisms: from diagnosis to treatment]. AB - Primary hyperaldosteronism is a growing cause of apparently essential hypertension (until 15 % of patients with hypertension). Primary hyperaldosteronism can now be recognized even for patients without hypertension or hypokaliemia. Paradoxically normal levels of aldosterone when serum renin levels and/or plasmatic renin activity and/or kaliemia are low, lead to suggest primary hyperaldosteronism. High values of aldosterone/renin or aldosterone/PRA favour the diagnosis. Adrenal scanography (fine slices) has to be performed, and confronted with one functional procedure (adrenal iodocholesterol uptake after dexamethasone, or sampling of adrenal venous blood for aldosterone measurement) only if surgery is suggested. When hypersecretion is unilateral or largely predominant in one size, surgery is very useful, when hypertension or hypokaliemia are poorly controlled, when medications are unwell tolerated, especially in men. PMID- 19299108 TI - Microbial antigens mediate HLA-B27 diseases via TLRs. AB - HLA-B27 positive individuals are predisposed to reactive arthritis developing 1-3 weeks after urogenital and gastrointestinal infections. Also ankylosing spondylitis (AS) associates strongly to HLA-B27, but no specific infection, Klebsiella pneumoniae excluded, has been linked to it. Before the discovery of its HLA-B27 association there were many reports suggesting a link between chronic prostatitis in men or pelvic inflammatory disease in women and AS. They have since been forgotten although HLA-B27 did not help to understand, why this disease has an axial and ascending nature. It is proposed that the urogenital organs form a source of damage (or danger)-associated molecular patterns (DAMPs), either exogenous pathogen-associated molecular patterns (PAMPs) from microbes or endogenous alarmins, such as uric acid, released from necrotic cells or urate deposits. DAMPs are slowly seeded from low-down upwards via the pelvic and spinal lymphatic pathways. They reach Toll-like receptors (TLRs) in their target mesenchymal stem cells, which are stimulated to ectopic enchondral bone formation leading to syndesmophytes and bamboo spine. At the same time inflammatory cytokines induce secondary osteoporosis of the spine. This new paradigm places microbes, HLA-B27 and TLRs in the pathogenic centre stage, but without pinpointing any (one) specific pathogen; instead, shared microbial patterns are indicated. PMID- 19299109 TI - Conjugated linoleic acid isomers' roles in the regulation of PPAR-gamma and NF kappaB DNA binding and subsequent expression of antioxidant enzymes in human umbilical vein endothelial cells. AB - OBJECTIVE: Conjugated linoleic acid (CLA) isomers have shown health benefits. Because CLA isomers may act as activators for peroxisome proliferator-activated receptors and may induce antioxidant enzymes, this study was conducted to examine the effects of CLA isomers on the gene expression of antioxidant enzymes, copper/zinc superoxide dismutase, and catalase in human umbilical vein endothelial cells. METHODS: Human umbilical vein endothelial cells were treated with graded concentrations of the 9-cis, 11-trans or the 10-trans, 12-cis-CLA isomer for 24 h. RESULTS: The 9-cis, 11-trans-CLA treatments resulted in increases in transcription factor DNA binding activities and expression of antioxidant enzymes at 0-25 micromol/L and an increase in lipid peroxidation only at the lowest concentrations (5 micromol/L). The 10-trans, 12-cis-CLA treatments resulted in increases in transcription factor DNA binding activities at 0-25 micromol/L and highest levels of mRNA of both antioxidant enzymes, superoxide dismutase protein, and lipid peroxidation only at the lowest concentrations (5 micromol/L). The 9-cis, 11-trans-CLA treatments produced expression of antioxidant enzymes, except catalase protein, that were positively correlated with lipid peroxidation. Positive correlations were found between expression of antioxidant enzymes, except catalase protein, and lipid peroxidation for 10 trans, 12-cis-CLA treatments. Although CLA isomers exhibit mostly stimulatory effects in expression of antioxidant enzymes, interestingly, the lowest concentrations of both CLA isomers resulted in increases in thiobarbituric acid reactive substance levels. CONCLUSION: An understanding of the optimal concentrations of CLA isomers, which stimulate the benefits of antioxidant enzyme induction, may require careful CLA titration to determine predictable and dependable therapeutic strategies against adverse effects, such as pro-oxidants. PMID- 19299110 TI - Safety of an IV formulation of carbamazepine. AB - An intravenous formulation of carbamazepine (CBZ) was administered to 113 (60 male; 53 female) persons with epilepsy aged 19-87 years. Subjects received 100mg of study drug as replacement for 100mg of their usual morning dose of CBZ. There were no significant changes in blood pressure or heart rate suggesting that this formulation can be developed as replacement therapy for persons unable to take oral CBZ. PMID- 19299111 TI - Hyperammonemia following intravenous valproate loading. AB - BACKGROUND: Valproic acid (VPA) has been associated with hyperammonemia with and without encephalopathy. We report the frequent but transient nature of hyperammonemia following intravenous (IV) administration of loading doses of VPA. METHODS: Forty participants received a VPA loading dose (20 or 30 mg/kg) at 6 or 10mg/kg/min. All participants were monitored for signs of systemic and local intolerance. Serum VPA level, ammonia, complete blood count, bilirubin, transaminases, pancreatic enzymes, and level of consciousness were obtained at baseline, 1 and 24h after administration. Changes in ammonia levels were assessed using repeated-measures ANOVA. RESULTS: Asymptomatic hyperammonemia occurred in 30 of 40 participants at 1h post-VPA infusion. Majority of the participants (66%) demonstrated decreasing ammonia concentrations at 24h post-infusion. Multivariable repeated-measures analysis indicates the lack of influence of VPA dose (p=0.8), VPA levels (p>0.24, all time points), infusion rate (p=0.41) and gender (0.68) on ammonia levels across time. Age (p=0.015), time since dosing (p=0.017) and co-therapy with enzyme-inducing antiepileptic drugs (p=0.035) were significant predictors of changes in ammonia levels. CONCLUSIONS: Hyperammonemia is a frequent but transient finding following intravenous administration of loading doses of VPA. Hyperammonemia was not associated with alteration in consciousness or hepatic transaminases. PMID- 19299112 TI - Transcriptional activity of genes coding transforming growth factor beta-1 and its receptors in patients with systemic sclerosis and Raynaud phenomenon. PMID- 19299113 TI - Anterior cingulate volume in adolescents with first-presentation borderline personality disorder. AB - Reports of volumetric abnormalities in the anterior cingulate cortex (ACC) in adults with established borderline personality disorder (BPD) are inconsistent, and it is not known whether such abnormalities are present early in the disorder. We aimed to investigate ACC volume in a first-presentation teenage BPD sample with minimal exposure to treatment. Fifteen female BPD patients and 15 healthy female control participants underwent magnetic resonance imaging (MRI) scanning. ACC volumes were estimated using a reliable method that accounts for inter individual variation in sulcal morphology, and measurements were compared between the two groups. Analysis of variance revealed a decrease in volume of the left ACC in BPD patients compared with control participants. This volumetric change was correlated with parasuicidal behavior and impulsivity. A measure of ACC volume asymmetry was also correlated with fear of abandonment symptoms. Our results suggest that ACC volumetric abnormalities early in the course of BPD might be related to clinical correlates of the disorder. Longitudinal studies are needed to examine the nature of this abnormality over the course of the disorder. PMID- 19299114 TI - Bulk and interfacial behaviour of caseinoglycomacropeptide (GMP). AB - Caseinoglycomacropeptide (GMP) is a hydrophilic glycopeptide released from milk kappa-casein by chymosin hydrolysis during cheese making. GMP is thought to be a potential ingredient for specific dietary applications with several health benefits. In this study GMP was characterized at the air-water interface and its behaviour was related with the self-assembly of GMP in solution as affected by pH. This GMP self-assembly was investigated by dynamic light scattering and the interfacial properties were determined by tensiometry and surface dilatational measurements at pH 4, 5 and 7. At pH 5 GMP exhibited higher surface pressure at equilibrium than at pH 7. At pH 4 the behaviour was more complex due to self assembly close to GMP pI. Dynamic measurement showed that the adsorption/penetration rate constant (K(ads)) is facilitated at higher GMP bulk concentrations, while the rate constant of rearrangement (K(r)) decreased at higher GMP concentrations which could be attributed to the existence of a steric restriction due to the higher GMP load at the interface. K(r) was higher at pH 5 because of lower electrostatic interactions close to the pI. The viscoelastic properties showed a complex behaviour due to the existence of protein-protein interactions depending on the GMP concentration, on the pH of the bulk and on the rates of diffusion, adsorption and rearrangement of GMP at the air-water interface. PMID- 19299115 TI - [Potential role of antiangiogenic treatment in neuroblastoma]. AB - Focus on new drug development over the last few years has yielded new agents that differ from unspecific classical chemotherapeutics and ionizing radiation, while still targeting the cancer cell itself. Antiangiogenesis is a totally distinct approach targeting the tumor's blood vessels. This concept has now found its eligibility for the treatment of several adult solid tumors: the human antivascular endothelial growth factor (VEGF) antibody bevacizumab, as well as the VEGF receptor tyrosine kinase inhibitors, sunitinib and sorafinib, have recently been licensed by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMEA) for the treatment of colorectal, renal, and lung cancer. Other antiangiogenic drugs are under preclinical and early clinical evaluation. However, what do we know of the use of these drugs in pediatric solid tumors, such as sarcomas and embryonal and neuronal tumors? For some time now, neuroblastoma has been shown to be dependent on angiogenesis. However, the first preclinical data on antiangiogenic drugs in neuroblastoma have not been published until recently, and clinical trials with antiangiogenic agents in neuroblastoma treatment protocols are scarce. This review adresses current knowledge on the important role and mechanisms of angiogenesis in neuroblastoma and summarizes available preclinical and clinical results of antiangiogenic agents used to treat neuroblastoma. Our review clearly demonstrates that clinical trials are urgently needed to bring forward promising antiangiogenesis concepts in neuroblastoma therapy. PMID- 19299116 TI - Long-term effects of St. John's wort (Hypericum perforatum) treatment: a 1-year safety study in mild to moderate depression. AB - Long-term safety and the effects of a St. John's wort (SJW) extract Ze 117 (Hypericum perforatum) were evaluated in the treatment of patients with depression. An open multicentre safety study with 440 out-patients suffering from mild to moderate depression according to ICD-10 was conducted. Patients were treated for up to 1 year with 500 mg St. John's wort extract per day (Ze 117). Evaluation criteria were safety (adverse event frequency) and influence on depression (HAM-D, CGI). Two hundred and seventeen (49%) patients reported 504 adverse events, 30 (6%) of which were possibly or probably related to the treatment. Gastrointestinal and skin complaints were the most common events associated with treatment. No age-related difference in the safety of the applied medication was found. The long-term intake of up to 1 year of the study medication did not result in any changes in clinical chemistry and electrocardiogram recordings. Body mass index (BMI) did not change either. Mean HAM-D scores decreased steadily from 20.58 at baseline to 12.07 at week 26 and to 11.18 at week 52. Mean CGI scores decreased from 3.99 to 2.20 at week 26 and 2.19 at week 52. Therefore, St. John's wort extract ZE 117 is a safe and effective way to treat mild to moderate depression over long periods of time, and therefore seems especially suitable for a relapse prevention. PMID- 19299117 TI - Kinome signaling through regulated protein-protein interactions in normal and cancer cells. AB - The flow of molecular information through normal and oncogenic signaling pathways frequently depends on protein phosphorylation, mediated by specific kinases, and the selective binding of the resulting phosphorylation sites to interaction domains present on downstream targets. This physical and functional interplay of catalytic and interaction domains can be clearly seen in cytoplasmic tyrosine kinases such as Src, Abl, Fes, and ZAP-70. Although the kinase and SH2 domains of these proteins possess similar intrinsic properties of phosphorylating tyrosine residues or binding phosphotyrosine sites, they also undergo intramolecular interactions when linked together, in a fashion that varies from protein to protein. These cooperative interactions can have diverse effects on substrate recognition and kinase activity, and provide a variety of mechanisms to link the stimulation of catalytic activity to substrate recognition. Taken together, these data have suggested how protein kinases, and the signaling pathways in which they are embedded, can evolve complex properties through the stepwise linkage of domains within single polypeptides or multi-protein assemblies. PMID- 19299118 TI - Engineered luciferases for molecular sensing in living cells. AB - As a means for visualizing molecular physiology within living cells, new strategies are emerging for engineering luciferases into intracellular biosensors. These biosensors can be classified as bimolecular, relying on complementation of luciferase fragments, or intramolecular, relying on domain insertion within the luciferase structure. Multiple design strategies have recently surfaced for the development of intramolecular sensors, allowing dynamic detection of small molecules or post-translational modifications within cells. Building on successes achieved in cell culture, these sensors are now beginning to reveal molecular processes within living organisms. PMID- 19299119 TI - Characterization of human epigenomes. AB - Histone modifications play a key role in regulating transcription and thus ultimately regulate cellular development and differentiation. To understand how histone modifications influence normal development and disease states, a global catalogue of histone modifications and modifying enzymes in normal and disease states is necessary. The first such systematic mapping experiments using the recently developed ChIP-Sequencing technique have revealed a combinatorial modification 'backbone' consisting of multiple histone modifications associated with active transcription. The human epigenomic datasets that are now being produced provide valuable resources for a better understanding of the functional regulatory elements of transcription and the pathways necessary for normal cellular development and pathological conditions. PMID- 19299120 TI - Doing Socrates experiment right: controlled rearing studies of geometrical knowledge in animals. AB - The issue of whether encoding of geometric information for navigational purposes crucially depends on environmental experience or whether it is innately predisposed in the brain has been recently addressed in controlled rearing studies. Non-human animals can make use of the geometric shape of an environment for spatial reorientation and in some circumstances reliance on purely geometric information (metric properties and sense) can overcome use of local featural information. Animals reared in home cages of different geometric shapes proved to be equally capable of learning and performing navigational tasks based on geometric information. The findings suggest that effective use of geometric information for spatial reorientation does not require experience in environments with right angles and metrically distinct surfaces. PMID- 19299121 TI - Endometriosis related to family history of malignancies in the Yale series. AB - OBJECTIVE: Recent studies reported that endometriosis could behave as a neoplasmatic process. The purpose of this study is to investigate the family risk of ovarian, colon and prostate cancer in women with endometriosis. STUDY DESIGN: A search of medical records at the Yale New Haven Hospital from 1996 to 2002 identified 348 women with endometriosis and 179 women without endometriosis. All the cases were diagnosed by laparoscopy. Demographic characteristics were evaluated in women with positive or negative family history of cancers in women with endometriosis. RESULTS: The overall risk of patients with endometriosis and positive family history of cancers was 7.7 (95% confidence interval 3.8-15.7) (chi(2)=39.8, P<0.001). Significant excess was observed for ovarian cancer in first- and second-degree relatives (OR=10.5, 95% CI (2.5-44.2), chi(2)=14.3, P<0.001), colon cancer (OR=7.5, 95% CI (2.7-21.1), chi(2)=18.2, P<0.001) and prostate cancer (OR=4.5, 95% CI (14-15.3), chi(2)=6.1, P<0.001). We found similar results in first- and second-degree relatives with ovarian and colon cancer. Moreover, we found similar results regarding the demographic characteristics in women with positive family history of cancers and in women with negative history. CONCLUSIONS: These data suggest a familial association of endometriosis with ovarian, colon and prostate cancers. This evidence could support the genetics and molecular similarities between endometriosis and cancer. Future studies will be important to determine a clear genetic link between endometriosis and cancer. PMID- 19299122 TI - Perineural and lymphovascular invasion on prostatic biopsy: pathological assessment and significance. AB - The importance of Gleason grade on prostatic needle biopsy specimens is widely accepted as a significant histopathological prognostic factor in the assessment of prostatic carcinoma. However, the value of other pathological variables used in the reporting of prostatic needle biopsies such as perineural or lymphovascular invasion is more controversial. This review examines the evidence for the utility of these pathological factors as prognostic indicators for prostatic carcinoma and the limitations in their assessment on needle biopsies. PMID- 19299123 TI - Antioxidant properties of a new antioxidative peptide from algae protein waste hydrolysate in different oxidation systems. AB - Microalgae have been a popular edible food, but there are no known reports on the antioxidative peptides derived from microalgae. The algae protein waste, which is normally discarded as animal feed, is a by-product during production of algae essence from microalgae, Chlorella vulgaris. Algae protein waste was hydrolyzed using pepsin, and a potent antioxidative peptide of VECYGPNRPQF was separated and isolated. The peptide could efficiently quench a variety of free radicals, including hydroxyl radical, superoxide radical, peroxyl radical, DPPH radical and ABTS radicals, and performed more efficiently than that observed for BHT, Trolox and peptides from marine protein sources in most cases. The purified peptide also has significant protective effects on DNA and prevents cellular damage caused by hydroxyl radicals. In addition, the peptide has gastrointestinal enzyme resistance and no cytotoxicity observed in human lung fibroblasts cell lines (WI 38) in vitro. These results demonstrate that inexpensive algae protein waste could be a new alternative to produce antioxidative peptides. PMID- 19299124 TI - Isolation of Pseudomonas stutzeri in wastewater of catfish fish-ponds in the Mekong Delta and its application for wastewater treatment. AB - The aim of this study was to explore the potential for reducing soluble N load in fishpond wastewater using naturally occurring denitrifying bacteria. Twenty-seven isolates were selected from in wastewater (liquid/solid) of catfish-ponds located along the Tien river, in the Mekong Delta, Vietnam in SW-LB medium (artificial seawater Luria-Britani medium) supplemented with 10 mM NH4 and NO3 and twenty five isolates were identified as Pseudomonas stutzeri based on similarity of PCR 16S rRNA using universal primers and specific primers. Four isolates were effective in lowering soluble N (NH4, NO2 and NO3) levels in fishpond water from 10 mg/L to negligible amounts after four days. Further experiments are underway to determine the fate of N lost from solution and the relative activity of ammonia oxidation, and nitrite and nitrate reduction by P. stutzeri isolates. PMID- 19299125 TI - Mediator facilitated, laccase catalysed oxidation of granular potato starch and the physico-chemical characterisation of the oxidized products. AB - The primary hydroxyl groups in potato starch were selectively oxidized to the corresponding aldehyde and carboxylic acid functionalities by mediators like TEMPO, using laccase from fungi as catalytic oxidant and oxygen as the primary oxidant. Oxidized starch products with degree of substitution (DS(CHO) ranging from 0.16 to 16.4/100AGU and DS(COOH) from 0.01 to 3.71 carboxyl groups/100AGU) were obtained with mediator facilitated enzymatic oxidation. Maximum conversion of the primary alcohol group was obtained at a pH of 5, with TEMPO as mediator, under oxygen bubbling and two step administration of Trametes versicolor laccase (200+200 nkat/g of starch). The oxidized products were characterised by IR spectroscopy, XRD and thermal studies. In the oxidized samples, the larger starch granules exhibited cracks and fractures in comparison to the smaller granules which were relatively unaffected, as observed from the microstructural studies. PMID- 19299126 TI - 2-Azidoalkoxy-7-hydro-8-oxoadenine derivatives as TLR7 agonists inducing dendritic cell maturation. AB - The synthesis of an array of 2-azidoalkoxy substituted 7-hydro-8-oxoadenines is described. The relation of the structure of these compounds and their ability to induce maturation of dendritic cells is evaluated. PMID- 19299127 TI - Exploring a pocket for polycycloaliphatic groups in the CXCR3 receptor with the aid of a modular synthetic strategy. AB - A CXCR3 pocket capable of accommodating polycycloaliphatics was explored using a modular synthetic strategy. The systematic studies reveal that the tricyclic 2 adamantane and bicyclic (iso)bornyl group are efficiently recognized by CXCR3. PMID- 19299128 TI - Design and synthesis of isoform-selective phospholipase D (PLD) inhibitors. Part II. Identification of the 1,3,8-triazaspiro[4,5]decan-4-one privileged structure that engenders PLD2 selectivity. AB - This Letter describes the synthesis and structure-activity relationships (SAR) of isoform-selective PLD inhibitors. By virtue of the installation of a 1,3,8 triazaspiro[4,5]decan-4-one privileged structure, PLD inhibitors with nanomolar potency and an unprecedented 40-fold selectivity for PLD2 over PLD1 were developed. Interestingly, SAR for this diverged from our earlier efforts, and dual PLD1/2 inhibitors were also discovered within this series. PMID- 19299129 TI - Synthesis and biological evaluation of new enantiomerically pure azole derivatives as inhibitors of Mycobacterium tuberculosis. AB - A series of novel enantiomerically pure azole derivatives was synthesized. The new compounds, bearing both an imidazole as well as a triazole moiety, were evaluated as antimycobacterial agents. One of them proved to have activity against Mycobaterium tuberculosis comparable to those of the classical antibacterial/antifungal drugs Econazole and Clotrimazole. PMID- 19299131 TI - Culicoides and the emergence of bluetongue virus in northern Europe. AB - In June 2006, bluetongue virus, an arboviral pathogen of ruminants, appeared in northern Europe for the first time, successfully overwintered and subsequently caused substantial losses to the farming sector in 2007 and 2008. This emergence served as a test of how the probability of arboviral incursion into new regions is assessed and has highlighted the reliance of decision making on paradigms that are not always underpinned by basic biological data. In this review, we highlight those areas of the epidemiology of bluetongue that are poorly understood, reflect upon why certain vital areas of research have received little attention and, finally, examine strategies that could aid future risk assessment and intervention. PMID- 19299132 TI - Fungal LysM effectors: extinguishers of host immunity? AB - Lysin motifs (LysMs) have been recognized in prokaryotes and plants as carbohydrate-binding protein modules. Recently, a novel virulence factor with LysMs was characterized from the plant pathogenic fungus Cladosporium fulvum. Here, we present a survey of public sequence data of 70 fungal species to demonstrate that putatively secreted LysM-containing proteins are widespread in the fungal kingdom, as they are found in mammalian and plant pathogenic species, in addition to saprophytes. We propose that these putative LysM effectors might have a role in sequestration of chitin oligosaccharides - breakdown products of fungal cell walls that are released during invasion and act as triggers of host immunity - to dampen host defence. PMID- 19299133 TI - Vesicular transport across the fungal cell wall. AB - Recent findings indicate that fungi use vesicular transport to deliver substances across their cell walls. Fungal vesicles are similar to mammalian exosomes and could originate from cytoplasmic multivesicular bodies. Vesicular transport enables the export of large molecules across the cell wall, and vesicles contain lipids, proteins and polysaccharides, many of which are associated with virulence. Concentration of fungal products in vesicles could increase their efficiency in food acquisition and/or delivering potentially noxious substances to other cells, such as amoebae or phagocytes. The discovery of vesicular transport in fungi opens many new avenues for investigation in basic cell biology and pathogenesis. PMID- 19299134 TI - Secretion and subcellular localizations of bacterial proteins: a semantic awareness issue. AB - Extracellular proteins are a subject of intense interest because of their essential roles in bacterial lifestyles. However, several terms related to secretion are used confusingly in the literature, and this is a topical issue in genomics and proteomics. Defining a secreted protein as actively translocated via a secretion system, here, we put into perspective that homologous translocation systems can result in radically different subcellular localizations of a secreted protein. We propose using standardized nomenclature for secretion systems from type I to type VIII for Gram-negative bacteria only, whereas the terms 'Sec' (secretion), 'Tat' (twin-arginine translocation), 'FEA' (flagella export apparatus), 'FPE' (fimbrilin-protein exporter), 'holin' (hole forming) and 'Wss' (WXG100 secretion system) should be applied to translocation systems across the cytoplasmic membrane of both Gram-positive and Gram-negative bacteria. Finally, we discuss why the term 'exoproteome' should be favoured over 'secretome' when describing the subset of proteins present in the extracellular milieu. PMID- 19299135 TI - Small RNAs and the control of transposons and viruses in Drosophila. AB - RNA interference (RNAi) - post-transcriptional gene silencing guided by small interfering RNA (siRNA) - is an important antiviral defense mechanism in insects and plants. Several recent studies in Drosophila identified endogenous siRNAs corresponding to transposons, to structured cellular transcripts and to overlapping convergent transcripts. In addition, one of these studies detected a large pool of Argonaute-2 associated siRNAs that mapped to the genome of flock house virus, a (+) RNA virus. Our bioinformatic analyses indicate that these viral siRNAs mapped in roughly equal proportions to both (+) and (-) viral RNA strands. These reports attribute an important function to RNAi in the defense against parasitic nucleic acids (viruses and transposable elements) and provide a novel mechanism for RNAi-based regulation of cellular gene expression. Furthermore, the detection of viral siRNAs of both (+) and (-) polarity implicates double-stranded RNA replication intermediates as the Dicer substrates that mediate antiviral defense. PMID- 19299136 TI - An atypical case of head trauma with late onset of contrecoup epidural hematoma, cerebellar contusion, and cerebral infarction in the territory of the recurrent artery of Heubner. AB - We encountered a case of head trauma with very unusual delayed events. A 68-year old woman was admitted to our hospital after receiving a direct impact to her occiput in a traffic accident. Head CT scans showed a thin acute epidural hematoma in the posterior fossa corresponding to a linear fracture, followed by late onset of contrecoup left frontal epidural hematoma and subsequent cerebellar contusion in the right cerebellar hemisphere. Fifteen days after the trauma, the patient developed mild motor weakness of right upper extremity. MRIs demonstrated an infarct in the territory of the left recurrent artery of Heubner. Although rare, atypical late events in patients with head trauma as reported here should be taken into consideration during subacute follow-up periods. PMID- 19299137 TI - A Chinese pedigree of cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL): clinical and radiological features. AB - Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is a rare hereditary cerebral vascular disease thought to be confined to Japan. This paper reports a Chinese pedigree of CARASIL in which two patients exhibited all of the typical clinical features of the disease. The radiological features are also discussed and compared with those of CADASIL. These cases illustrate the unique clinical and radiological features of CARASIL and challenge the idea that CARASIL is limited to the Japanese population. PMID- 19299138 TI - Primary cystic germinoma originating from the midbrain. AB - A primary intracranial germinoma that involves the midbrain is rare. We describe an unusual case of primary cystic germinoma originating from the midbrain. A 29 year-old man presented with diplopia, and his MRI showed a cystic, ring-like enhanced lesion in the thalamo-mesencephalic junction. Open biopsy was performed and the diagnosis of germinoma was based upon the histopathological findings. Following chemotherapy and radiotherapy, the symptoms improved and the tumor disappeared. We propose that primary intracranial germinoma should be included in the differential diagnosis of midbrain tumors, because early diagnosis and appropriate treatment for midbrain germinoma improves clinical outcome. PMID- 19299139 TI - Preliminary comparison of radiolucent cages containing either autogenous cancellous bone or hydroxyapatite graft in multilevel cervical fusion. AB - We compared the preliminary outcomes of cervical fusion performed using radiolucent cages containing either cancellous bone or hydroxyapatite graft. From July 2004 to June 2006, 45 consecutive patients presented with a total of 109 levels of degenerative disc disease between the C2 and C7 levels. Each patient underwent anterior cervical discectomy and fusion (ACDF) for each affected cervical disc. The retrospective analysis of the cage fillers was divided into group 1 (23 patients with 56 affected cervical levels) who received cages packed with cancellous bone marrow, and group 2 (22 patients with 53 affected cervical levels) who received cages packed with hydroxyapatite graft. Bone marrow was harvested from the anterior iliac crest. The Prolo scale was used to assess both the economic and functional status postoperatively. The Yates' correction to test independence in a contingency was used to compare the fusion rate of both groups post-operatively at day 1 and at 1, 3 and 6 month follow-up. At a mean follow-up of 12 months, the fusion rates observed in groups 1 and 2, respectively, were 21.4% and 13.2% after 1 month, 76.8% and 64.2% after 3 months, and 98.2% and 96.2% after 6 months. Functional and economic status were better in group 2, with a statistical significance (p<0.05) observed at the 3-month follow-up. Although hydroxyapatite graft is an osteoconductive, rather than osteoinductive, material, when used as a cage filler it is a safe and efficient substitute for cancellous bone. PMID- 19299140 TI - Clinical features and surgical outcomes of ruptured distal anterior cerebral artery aneurysms in 20 consecutively managed patients. AB - Optimal surgical management of ruptured aneurysms of the distal anterior cerebral artery continues to provide unique technical challenges. This study presents 20 consecutively managed such patients, with special attention given to the clinical and radiological characteristics, as well as the surgical outcomes. Intracerebral hematoma was seen in 11 of the 20 patients, and intraventricular hemorrhage occurred in 4 (20%). Angiography revealed that 9 (45%) patients had multiple aneurysms. Three patients (15%) had "mirror" distal anterior cerebral arterial aneurysms on the contralateral side. Eleven patients (55%) had aneurysms located at the supracallosal portion of the anterior cerebral artery, while 9 patients (45%) had aneurysms located below the genu of the corpus callosum. The mean aneurysmal diameter was 3.85 mm; 18 aneurysms (90%) were less than 6 mm in diameter. Eighteen patients (90%) underwent a microsurgical procedure; 2 (10%) underwent endovascular coiling due to poor clinical grade. A favorable outcome was achieved in 14 (70%) patients. Advances in microsurgical techniques will be required to further improve clinical outcome. PMID- 19299141 TI - The ribozyme core of group II introns: a structure in want of partners. AB - Group II introns contain a large ribozyme, which catalyzes self-splicing, and the coding sequence of a reverse transcriptase, the function of which is to cooperate with the ribozyme to achieve genomic mobility. Despite its lack of substrates for both steps of the splicing process, the crystal structure of a group II ribozyme reveals the location of two metal ions most likely to be involved in catalysis; the RNA structure that binds to these ions results from the bending of a local motif by the folding of the rest of the ribozyme. The stage is now set to determine where the intron-encoded protein binds to its partner and whether the spliceosome uses a counterpart of the group II catalytic center to excise nuclear pre-messenger introns. PMID- 19299142 TI - On the origin of the cap-dependent initiation of translation in eukaryotes. AB - The Shine-Dalgarno sequence of prokaryotic mRNAs, which helps to bind and position the ribosome at the start site for protein synthesis, is absent from eukaryotic mRNAs. Instead, for most, a structure at the 5' end and a much larger number of protein initiation factors are needed for both binding of the ribosome and for successful start-site selection, that is, a 'cap-dependent' initiation mechanism. Although the mechanics of this process are well studied, what is not clear is how it evolved. By analyzing recent progress in different fields, I suggest that it was the need to adjust to the arrival of the nuclear membrane and the subsequent requirement to export intron-less mRNAs to the cytoplasm that spurred the shift to the more complex translation initiation mechanism in eukaryotes. PMID- 19299143 TI - Regulation of phosphoinositide 3-kinase expression in health and disease. AB - Both the biology and the therapeutic potential of the phosphoinositide 3-kinase (PI3K) signalling axis have been the subject of intense investigation; however, little is known about the regulation of PI3K expression. Emerging evidence indicates that PI3K levels change in response to cellular stimulation with insulin and nuclear receptor ligands, and during various physiological and pathological processes including differentiation, regeneration, hypertension and cancer. Recently identified mechanisms that control PI3K production include increased gene copy number in cancer, and transcriptional regulation of the p110alpha PI3K gene by FOXO3a, NF-kappaB and p53, and of the PI3K regulatory subunits by STAT3, EBNA-2 and SREBP. In most instances, however, the impact of alterations in PI3K expression on PI3K signalling and disease remains to be established. PMID- 19299144 TI - Revision total knee arthroplasty: the influence of femoral stems in load sharing and stability. AB - Restoration of lost bone support and joint stability are the primary challenges in revision total knee arthroplasty (TKA). Normally, the defects type 2B and type 3 are associated with several damaged metaphyseal bone and loss of cortical bone. Structural allografts have been used for the treatment of large, contained, or uncontained osseous defect in revision TKA. Disadvantages of using structural allografts include late resorption or nonunion and risk of disease transmission. Alone, the structural allograft can not provide the initial support for revision implants. In these cases the stems are frequently used to provide the necessary load sharing and increase initial component stability. When evaluating whether stems should be cemented or press-fit, there isn't a simple answer, since there are no clear advantages or disadvantages for each approach. The aim of this study was to evaluate load sharing and stability at bone graft-cement interface under femoral component with use of cemented and press-fit stems after allograft incorporation with host bone. The difficulties encountered or the impossibility on the evaluation of these biomechanical parameters in-vitro make the use of finite element (FE) models a way through this evaluation. The FE results suggest that a press-fit stem could be the adequate choice if structural allografts in femur revision are used. PMID- 19299145 TI - Efficacy of intra-articular cocktail analgesic injection in total knee arthroplasty - a randomized controlled trial. AB - In a randomized, double-blind, placebo, parallel and controlled study, 80 patients with osteoarthritis who underwent unilateral TKA were randomly assigned to two groups: Trial Group, where patients received intra-articular intraoperative injection containing morphine, bupivacaine and betamethasone, and Control Group, where patients received normal saline as control. All patients received patient-controlled analgesia (PCA) for 48 h postoperatively. We found that intra-articular cocktail analgesic injection significantly reduced the morphine consumption during the 0-36 h postoperative period and the total morphine consumption. VAS at rest in Trial Group at postoperative 6, 10, 24 and 36 h was significantly lower than that in Control Group, and VAS during activity in Trial Group at postoperative 24 h and 36 h was significantly lower than that in Control Group. The time of ability to perform an active straight leg raise and to actively reach 90 degrees knee flexion, as well as ROM of the knee at the 15th postoperative day, was better in Trial Group than those in Control Group. There were no significant differences in postoperative wound healing, infection, blood pressure, heart rate, rash, respiratory depression, urine retention and DVT between the two groups. The occurrence of nausea and vomiting in Trial Group was lower than that of Control Group. This study revealed that intra-articular cocktail analgesic injection reduced the need for morphine and offered a better pain control, without apparent risks following TKA. PMID- 19299146 TI - Design, synthesis and SAR studies of tripeptide analogs with the scaffold 3 phenylpropane-1,2-diamine as aminopeptidase N/CD13 inhibitors. AB - Aminopeptidase N (APN), belonged to metalloproteinase, is an essential peptidase involved in the process of tumor invasion and metastasis. A series of tripeptide analogs with the scaffold 3-phenylpropane-1,2-diamine were designed, synthesized and evaluated for their ability to inhibit APN. Preliminary activity evaluation showed that most of target compounds possessed potent inhibitory activities against APN. With in this series, compound A6 and B6 exhibited good potency with the IC(50) values of 8.8+/-1.3 microM and 8.6+/-1.1 microM, respectively. PMID- 19299147 TI - Fluorinated isatin derivatives. Part 1: synthesis of new N-substituted (S)-5-[1 (2-methoxymethylpyrrolidinyl)sulfonyl]isatins as potent caspase-3 and -7 inhibitors. AB - A series of new N-substituted (S)-5-[1-(2 methoxymethylpyrrolidinyl)sulfonyl]isatin derivatives has been synthesized and tested as inhibitors of caspases-3 and -7, which are known to be downstream enzymes critical in the execution of apoptosis. N-Propyl- and N-butyl isatins, as well as the corresponding terminal alcohols and regioisomeric fluorobutyl derivatives were shown to be excellent inhibitors having different binding potencies for caspases-3 and -7. In contrast, the corresponding fluoroethyl and fluoropropyl compounds were about 100-1000 times less active. Fluorinated N benzyl isatins as well as trifluoroalkyl and difluoroalkyl derivatives were moderate inhibitors. However, isatins bearing different alkylether groups at N-1 are very weak or not active as inhibitors of caspases-3 and -7. PMID- 19299148 TI - Antimalarials from nature. AB - Malaria is a major public health problem mainly due to the development of resistance by the most lethal causative parasitic species, Plasmodium falciparum to the mainstay drugs like chloroquine. New drugs with unique structures and mechanism of action are urgently required to treat sensitive and drug-resistant strains of malaria. Historically, compounds containing novel structure from natural origin represent a major source for the discovery and development of new drugs for several diseases. This review presents recent advances in antimalarial drug discovery from natural sources, including plant extracts, and compounds isolated from plants, bacteria, fungi and marine organisms. These compounds offer new and novel scaffolds for development as antimalarials. The literature from 1998 to October 2008 is reviewed. The review present literature compilation from plant and marine extracts, alkaloids (naphthylisoquinolines, bisbenzylisoquinolines, protoberberines and aporphines, indoles, manzamines, and miscellaneous alkaloids) terpenes (sesquiterpenes, triterpenes, diterpenes, and miscellaneous terpenes) quassinoids, flavonoids, limonoids, chalcones, peptides, xanthones, quinones and coumarines, and miscellaneous antimalarials from nature. The review also provides an outlook to recent semisynthetic approaches to antimalarial drugs discovered from natural sources. PMID- 19299149 TI - Nanoparticles in cellular drug delivery. AB - This review highlights the properties of nanoparticles used in targeted drug delivery, including delivery to cells as well as organelle targets, some of the known pharmacokinetic properties of nanoparticles, and their typical modifications to allow for therapeutic delivery. Nanoparticles exploit biological pathways to achieve payload delivery to cellular and intracellular targets, including transport past the blood-brain barrier. As illustrative examples of their utility, the evaluation of targeted nanoparticles in the treatment of cancers and diseases of the central nervous system, such as glioblastoma multiforme, neurovascular disorders, and neurodegenerative diseases, is discussed. PMID- 19299150 TI - Evidence of multi-walled carbon nanotube fragmentation induced by sonication during nanotube encapsulation via bulk-suspension polymerization. AB - The synthesis of multi-walled carbon nanotube/polystyrene composites, with nanotube concentrations of 0.04, 0.08 and 0.16 wt%, was carried out by in situ bulk-suspension polymerization with the assistance of sonication. By using this method both encapsulation and exfoliation of the nanotubes into the polymer host were achieved. Evidence of significant nanotube fragmentation was found by scanning electron microscopy; the cause of such fragmentation was attributed to the induction of strong cavitation due to the application of ultrasound during the synthesis. Infrared spectroscopy showed no evidence of the formation of covalent bonds between the nanotubes and the polystyrene during the process of synthesis. The thermal stability was not improved by the inclusion of the nanotubes, it was attributed to the low nanotube concentrations; however, composites glass transition temperature showed improvements. PMID- 19299151 TI - Implementation of the ICRP 2007 recommendations in Korea. AB - The International Commission on Radiological Protection (ICRP) published the new recommendations 2007 on radiological protection. International Atomic Energy Agency (IAEA) is also under process in revising its International Basic Safety (BSS). According to revision of the ICRP recommendations and BSS, the Korean government plans to implement those changes. In the 2007 ICRP recommendations, there are some new concepts, principles and quantities. Based on the study carried out by Korea Institute of Nuclear Safety (KINS) so far, the following points are identified as major areas that need further in-depth review and consideration for the implementation of the ICRP 2007 recommendations into Korean radiation protection laws and regulations; changes in the radiation risk factors, radiation weighting factors and tissue weighting factors, maintenance of the ICRP 60 dose limits, practical application of the dose constraints and determination of the reference levels in many source to individual exposure relationships, change from process-based system to exposure situation-based system, strengthening of the principle of optimization in all exposure situations, system of radiation protection for the environment, practical application of the exclusion and exemption principles, active participation of the stakeholders, changes in glossary etc. The study for the implementation of the ICRP 2007 recommendations into national legislations will be conducted till the end of 2012. In the meantime, draft regulations will be developed and the possible impact on the nuclear industry will also be analyzed and active involvement of the stakeholders including licensees will be encouraged in the entire process. The final draft of the revised laws and regulations will be issued in the early of 2013 and the formal legislation process of this final draft will commence in due course. PMID- 19299152 TI - Fabrication and performance characteristics of a CsI(Tl)/PIN diode radiation sensor for industrial applications. AB - CsI(Tl)/PIN diode radiation sensors were fabricated for application in various fields such as an NDT and an environmental radiation monitoring system. CsI(Tl) crystals of 11x11x21mm(3) were processed as optical grade from a CsI(Tl) ingot and matched with PIN diodes in consideration of the light loss and the external impact. The photodiode signal is amplified by a low-noise preamplifier and a pulse shape amplifier. At room temperature, the fabricated CsI(Tl)/PIN diode radiation sensors demonstrate an energy resolution of 7.9% for 660keV gamma rays and 4.9% for 1330keV. The fluctuation of the directional dependency was below 14% from 0 to 90 degree for the incident 660keV gamma rays. The compactness, the low voltage power supply and the physical hardness are very useful features for industrial applications of the fabricated CsI(Tl)/PIN diode sensor. PMID- 19299154 TI - Regulatory implications of a linear non-threshold (LNT) dose-based risks. AB - Current radiation protection regulatory limits are based on the linear non threshold (LNT) theory using health data from atomic bombing survivors. Studies in recent years sparked debate on the validity of the theory, especially at low doses. The present LNT overestimates radiation risks since the dosimetry included only acute gammas and neutrons; the role of other bomb-caused factors, e.g. fallout, induced radioactivity, thermal radiation (UVR), electromagnetic pulse (EMP), and blast, were excluded. Studies are proposed to improve the dose response relationship. PMID- 19299153 TI - A simple Cu-64 production and its application of Cu-64 ATSM. AB - One of the positron emission radionuclides, (64)Cu, has been reported to be a particularly effective radioisotope in PET imaging study. This utility of (64)Cu depends on the chemical stability in water with proper energy and half-life as gamma-emitters. Hence, we tried to develop a simple method for producing this isotope using an old cyclotron model in our site (50MeV, Scantronics co.). In particular, we designed the equipments of enrich (64)Ni plating system and radioactive (64)Cu separation using plastic cartridge column; (64)Ni plating system on gold foil which located in the 13 degrees angle target toward beam irradiation. For the nuclear reaction of (64)Cu, it was applied to (64)Ni(p, n) (64)Cu at low energy under the degrader composed of Al and Ta foils. After beam irradiation, (64)Cu was identified by multichannel analyzer installed for a HPGe detector and its utility was certified by the microPET images of (64)Cu-ATSM (CT 26 tumor bearing mouse as reported previously). The image quality of (64)Cu was also very similar to that of (18)F radioisotope in microPET scanner. In conclusion, a method of (64)Cu production and its application was successfully established in old cyclotron having high energy. PMID- 19299155 TI - Development of a method for activity measurements of 232Th daughters with a multidetector gamma-ray coincidence spectrometer. AB - The method for activity measurements of the (232)Th daughters, developed at the six-crystal gamma-ray coincidence spectrometer PRIPYAT-2M and based on coincidence counting of the 583 and 2615 keV photons from cascade transitions which follow beta(-)-decay of (208)Tl, as well as on counting the 911 keV photons which follow beta(-)-decay of (228)Ac in the integral and non-coincidence mode of counting, is presented. PMID- 19299156 TI - The production of (88)Y in the proton bombardment of (nat)Sr: New excitation and separation studies. AB - The cyclotron production of (88)Y at iThemba LABS is performed via the reaction (88)Sr(p,n)(88)Y. The yields obtained were inconsistent with nuclear data obtained from the literature and the excitation function of the nuclear reaction was re-measured, using a differentiation of thick-target production rate measurements. Ion exchange chromatographic methods are described to separate (88)Y from (nat)Sr target material using AG MP-1 resin and AG 50W-X4 resins, respectively. PMID- 19299157 TI - Data analysis from monitoring of radionuclides in the nuclear power plant Temelin ecosystem area. AB - Changes in the ecosystem occur naturally, however, can also be due to man's activity. The unique ecosystem monitoring based on studying contamination of the bioindicators by man-made radionuclides has been providing data from the area of nuclear power plant (NPP) Temelin (CR) using laboratory gamma spectrometry for past 8 years. The increase in radioactivity in the NPP ecosystem area resulting from its operation is evaluated through comparison with a zero level of (137)Cs mass activities and by obtained data trend analysis. PMID- 19299158 TI - Conceptual design of experimental equipment for large-diameter NTD-Si. AB - An irradiation-experimental equipment for 12in neutron transmutation doping silicon (NTD-Si) was designed conceptually by using MCNP5 in order to improve the neutron flux distribution of the radial direction. As a result of the calculations, the neutron absorption reaction ratio of the circumference to the center could be limited within 1.09 using a thermal neutron filter that covers the surface of the silicon ingot. The uniformity of the (30)Si neutron absorption was less than 5.3%. PMID- 19299159 TI - Environmental mobility of cobalt-Influence of solid phase characteristics and groundwater chemistry. AB - The adsorption of cobalt on samples from a potential waste repository site in an arid region was investigated in batch experiments, as a function of various solution phase parameters including the pH and ionic strength. The samples were characterized using a range of techniques, including BET surface area measurements, total clay content and quantitative X-ray diffraction. The statistical relationships between the measured cobalt distribution coefficients (K(d) values) and the solid and liquid phase characteristics were assessed. The sorption of cobalt increased with the pH of the aqueous phase. In experiments with a fixed pH value, the measured K(d) values were strongly correlated to the BET surface area, but not to the amount of individual clay minerals (illite, kaolinite or smectite). A further set of sorption experiments was undertaken with two samples of distinctive mineralogy and surface area, and consequently different sorption properties. A simple surface complexation model (SCM) that conceptualized the surface sites as having equivalent sorption properties to amorphous Fe-oxide was moderately successful in explaining the pH dependence of the sorption data on these samples. Two different methods of quantifying the input parameters for the SCM were assessed. While a full SCM for cobalt sorption on these complex environmental substrates is not yet possible, the basic applicability and predictive capability of this type of modeling is demonstrated. A principal requirement to further develop the modeling approach is adequate models for cobalt sorption on component mineral phases of complex environmental sorbents. PMID- 19299160 TI - Neurectomy of the trigeminal nerve branches: clinical evaluation of an "obsolete" treatment. AB - INTRODUCTION: After all options of conservative treatment of trigeminal neuralgia have been exhausted, surgical methods are used. Peripheral neurectomy is one of the simplest among them. It is a relatively obsolete method, but can still be useful and effective. AIMS: To analyse the remission period after repetitive neuroexeresis of the same neural branch, to calculate "predicted" remission, and to evaluate the benefits of repetitive neuroexeresis. PATIENTS AND METHODS: Total of 85 neurectomies were performed in 36 patients in 5 years by intraoral access. The recurrence time was analysed by simple regression analysis, using the method of the least-squares. Thus, the average pain free time after repetitive neurectomy can be predicted. RESULTS: After the first operation on the infraorbital nerve, recurrence mainly appeared in the period between 12 and 15 months, after the second between 9 and 12 months. After the first operation on the mandibular nerve, recurrence took place mainly after 15-18 months, after the second 12.3 months on the average. No interventions were performed on the first branch. Calculated prediction of the average time of remission was in agreement with the actual duration of painlessness. CONCLUSION: Remission time after repetitive neurectomy decreases. There is no point in repeating the surgery on the same neural branch more than three times. PMID- 19299161 TI - Osseous alterations at the interface of hydrogel expanders and underlying bone. AB - INTRODUCTION: In plastic and reconstructive surgery, self-activating hydrogel expanders are used to augment soft-tissue space. The purpose of this study was to investigate the morphological response of underlying bone to the constant pressure exerted by a hydrogel expander. METHODS: Eighteen Lewis rats were randomly divided into three groups. In group 1, a hydrogel expander was placed subperiosteally directly onto the calvaria of the rats. In group 2, the expander and the underlying bone were separated by a polydioxanone (PDS) foil. Group 3 animals served as controls. Before and 14 days after the insertion of the expanders, micro-computed tomography (CT) images were obtained and fused. We analysed hydroxyapatite density beneath and at the periphery of the expander and performed a histomorphometric bone analysis. RESULTS: Whereas there were no significant differences (p<0.05) (groups 1 and 2) in bone density at the periphery of the expanders between the study groups, a significant decrease in hydroxyapatite density beneath the expanders was observed in those animals in which the devices were placed directly onto the calvaria (group 1). Whereas bone thickness was unaffected at the periphery of the expanders in all groups, it was significantly decreased beneath the expanders in all implanted animals. A morphological examination revealed resorption lacunae with a diameter of 218.4+/ 56 mICROm in those rats in which the expanders had been placed directly onto the calvaria. CONCLUSION: This study shows the direct influence of hydrogel expanders on underlying bone. Whereas bone resorption and connective tissue formation also occur underneath hydrogel expanders, these effects can be avoided if the expander and the underlying bone are separated by PDS foil. The key to success is to ensure the appropriate placement of expanders and thus to avoid bone resorption. PMID- 19299162 TI - Improving compliance and persistence to adjuvant tamoxifen and aromatase inhibitor therapy. AB - Better compliance and persistence with therapy are associated with improved patient outcomes. As more and more patients survive breast cancer, compliance with adjuvant therapy becomes increasingly important. In clinical trials, compliance with adjuvant endocrine therapy among women with breast cancer is usually high. Retrospective analyses of databases and medical records from clinical practice, insurance databases of prescription refills, and survey data show a significant decrease in persistence after 12 months of therapy. With ongoing therapy, a further decline in persistence of up to 50% has been reported. A consistent methodology is needed to measure patient behavior and identify patients who are not adhering to therapy. Promising strategies for enhancing adherence to treatment in clinical practice include improving access to health care, increasing patient satisfaction, managing side effects, patient education, and better communication between the patient and health care provider. Positive relationships between patients and their health care providers, and frequent monitoring and feedback, may be most effective. While the lack of conformity across studies in measuring makes cross-study comparisons difficult, this review evaluates the available data regarding compliance and persistence with adjuvant endocrine therapies for breast cancer (tamoxifen and aromatase inhibitors) and presents strategies for improving adherence. PMID- 19299163 TI - Induction of Th1 cytokines by Leuconostoc mesenteroides subsp. mesenteroides (KCTC 3100) under Th2-type conditions and the requirement of NF-kappaB and p38/JNK. AB - Leuconostoc mesenteroides subsp. mesenteroides (LMM) KCTC 3100, is one of the prominent species in the fermentation of kimchi, a traditional Korean food. In the present study, we investigated the capacity of this microorganism in inducing Th1 cytokines in the presence of Th2 signals in vitro and in vivo and the requirement of NF-kappaB and MAPK signaling. Stimulation with heat-killed LMM in mouse splenocytes induced the expression of IFN-gamma, which was dependent on IL 12 production by LMM. Pre-treatment with LMM in vitro augmented the production of IFN-gamma and IL-4 in response to anti-CD3 plus recombinant IL-4 (rIL-4). LMM administration to mice, beginning either before or after the development of OVA sensitization, increased OVA-restimulated IFN-gamma production in the splenocytes and reduced serum total and OVA-specific IgE levels. However, only the pre sensitization treatment induced a slight reduction in IL-4 from the same cells, but the post-sensitization treatment did not. Induction of IL-12 by LMM in peritoneal macrophages involved NF-kappaB, p38 and JNK, but not ERK1/2. In conclusion, our data presented the upregulation of IFN-gamma by LMM under the pro Th2 conditions and the requirement of NF-kappaB, p38 and JNK for IL-12 production. These observations suggest that this microorganism can be a useful Th1-inducing agent in modulating the Th1/Th2 imbalance. PMID- 19299164 TI - TLR2 sensing of F. nucleatum and S. sanguinis distinctly triggered gingival innate response. AB - Gingival tissue faces constant exposure to micro-organisms. It functions as part of the host response, an anti-microbial barrier that recognizes and discriminates between commensal and pathogenic bacteria. This study aimed to evaluate and compare the effects of cell wall extracts from different periodontal bacteria, commensals Streptococcus sanguinis and Fusobacterium nucleatum and the pathogen Porphyromonas gingivalis, on the innate immune response of gingival keratinocytes and the role of TLR2 in regulating this. We assayed mRNA levels to determine the expression of human beta-defensins (hbetaD2, hbetaD3), interleukin-1alpha, 1beta, 6 and 8 and matrix metalloproteinase-9. F. nucleatum extracts induced beta defensin and inflammatory marker mRNA expression at higher levels than P. gingivalis. Extracts from the Gram-positive commensal S. sanguinis did not upregulate the host response. TLR2 extinction inhibited the upregulation of beta defensin and cytokine transcripts by F. nucleatum extracts but, in contrast, led to a weak induction of hbetaD3 after challenge with S. sanguinis extracts. Although F. nucleatum strongly induces innate immune and inflammatory mediators, S. sanguinis limits their expression through TLR2. Together, our data demonstrate that gingival keratinocytes recognize and discriminate between Gram-positive and Gram-negative commensal extracts, in part through TLR2, to activate different signaling pathways of the innate immune host response. PMID- 19299165 TI - Comparison between vacuum sublimed matrices and conventional dried droplet preparation in MALDI-TOF mass spectrometry. AB - The properties of several cinnamic acid compounds used as matrices for matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were investigated as standard dried droplet (DD) and vacuum sublimed preparations. The differences between both preparation methods were analyzed with regard to matrix grain size, internal ion energy, initial velocity, analyte intensity, and analyte incorporation depth. Some of the used cinnamic acid derivatives exhibit clearly reduced grain sizes as sublimed preparations compared with standard DD approaches. In these cases higher effective temperatures could be measured accompanied by increased analyte intensities, which can be explained by stronger volatilization processes caused by a hindered heat dissipation resulting in a raised analyte transfer into the gas phase. For all sublimed compounds, a strong increase of the initial ion velocity compared with DD preparations could be measured. Higher initial ion velocities correlate with a decrease in internal ion energy which might be attributed to the very uniform crystal morphology exhibited by sublimed compounds. For sublimed matrices without reduced grain size, at least slightly higher analyte intensities could be detected at raised laser fluences. Analyte accumulation in the uppermost matrix layers or the detected higher ion stability can be explanations for these results. PMID- 19299166 TI - Rephasing ion packets in the Orbitrap mass analyzer to improve resolution and peak shape. AB - A method is described to improve resolution and peak shape in the Orbitrap under certain experimental conditions. In these experiments, an asymmetric anharmonic axial potential was first produced in the Orbitrap by detuning the voltage on the compensator electrode, which results in broad and multiply split mass spectral peaks. An AC waveform applied to the outer electrode, 180 degrees out of phase with ion axial motion and resonant with the frequency of ion axial motion, caused ions of a given m/z to be de-excited to the equator (z = 0) and then immediately re-excited. This process, termed "rephasing," leaves the ion packet with a narrower axial spatial extent and frequency distribution. For example, when the Orbitrap axial potential is thus anharmonically de-tuned, a resolution of 124,000 to 171,000 is obtained, a 2- to 3-fold improvement over the resolution of 40,000 to 60,000 without rephasing, at 10 ng/microL reserpine concentration. Such a rephasing capability may ultimately prove useful in implementing tandem mass spectrometry (MS/MS) in the Orbitrap, bringing the Orbitrap's high mass accuracy and resolution to bear on both the precursor and product ions in the same MS/MS scan and making available the collision energy regime of the Orbitrap, approximately 1500 eV. PMID- 19299167 TI - Incoherent production reactions of positive and negative ions in matrix-assisted laser desorption/ionization. AB - Utilizing synchronized dual-polarity matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, we found good evidence of the incoherent production of positive and negative matrix ions. Using thin, homogeneous 2,5-dehydroxybenzoic acid (DHB) matrix films, positive and negative matrix ions were found to appear at different threshold laser fluences. The presence of molecular matrix ions of single charge polarity suggests that the existence of DHB ion-pairs may not be a prerequisite in MALDI. Photoelectrons induced by the laser excitation may assist the production of negative DHB ions, as shown in experiments conducted with stainless steel and glass substrates. At high laser fluences, the relative yield of positive and negative matrix ions remained constant when homogeneous matrix films were used, but it fluctuated significantly with inhomogeneous crystal morphology. This result is also inconsistent with the hypothesis that matrix ion pairs are essential primary ions. Evidence from both low and high laser fluences suggests that the productions of positive and negative matrix ions in MALDI may occur via independent pathways. PMID- 19299168 TI - A method to enhance a1 ions and application for peptide sequencing and protein identification. AB - A simple and effective method was developed for peptide sequencing and protein identification through the determination of its N-terminal residue. The method of N-terminal carbamidomethylation with iodoacetamide could specifically and remarkably enhance the intensity of a(1) ions in the tandem mass spectra of the peptide derivatives without significantly altering their fragmentation pattern, thus allowing determination of their N-terminal residues. The effectiveness and specificity of the method was demonstrated by confirming and extending sequence interpretation of several model peptides and proteins. The developed method was then applied in the LC-MS/MS analysis of the tryptic digests of myoglobin and a whole protein extract from rat heart tissues. The results from database searches were well validated with the enhancement of a(1) ions in tandem mass spectra and the specificity of protein identification was obtained when the information of N terminal residues was included in the database search. PMID- 19299169 TI - Proposed algorithm to investigate latent and occult viruses in vaccine cell substrates by chemical induction. AB - The recent urgency to develop new vaccines for emerging and re-emerging diseases, such as pandemic influenza, has necessitated the use of cell substrates not previously used in the manufacture of licensed vaccines. A major safety concern in the use of novel cell substrates is the presence of potential adventitious agents, such as latent and occult viruses, that may not be detected by currently used conventional assays. In cases where the novel cell substrate is known to be tumorigenic, there are additional safety issues related to tumorigenicity of intact cells and oncogenicity of residual cellular DNA. We have developed a strategy for evaluating vaccine cell substrates for the presence of latent/occult viruses, including endogenous retroviruses, latent RNA viruses and oncogenic DNA viruses, by optimizing conditions for chemical induction of viruses and using a combination of broad and specific assays to enable detection of known and novel viruses. PMID- 19299170 TI - Radiofrequency thermal ablation of small tumors in transplanted kidneys: an evolving nephron-sparing option. AB - Small tumors in transplanted kidneys are rarely detected during follow-up. With surgical treatment, it is sometimes challenging to preserve the graft and its function. Image-guided ablation techniques have been recently reported as effective in treating these tumors. Herein three cases of radiofrequency (RF) thermal ablation in transplanted kidneys are reviewed and compared with cases in the literature. There were no major complications seen, the mean change in serum creatinine level was no greater than 0.1 mg/dL, and complete ablation was sustained on imaging studies at 3, 18, and 62 months follow-up. RF ablation can be considered a safe, function-preserving, and effective evolving therapeutic option for small tumors in transplanted kidneys. PMID- 19299171 TI - Amylin reduces plasma glucagon concentration in cats. AB - Pancreatic amylin plays an important role in the control of nutrient fluxes and is an established therapy in human diabetes as it reduces post-prandial glucagon secretion and slows gastric emptying. Given the similar pathophysiology of human type-2 and feline diabetes mellitus, we investigated whether amylin reduces plasma glucagon levels in cats. Healthy cats were tested using an intravenous arginine stimulation test (IVAST), a meal response test with the test meal comprising 50% of average daily food intake, and an IV glucose tolerance test (IVGTT). Non-amyloidogenic rat amylin injected 5 min before the respective stimulus significantly reduced plasma glucagon levels under all test situations. In the IVAST and IVGTT, cats treated with amylin also had lower plasma insulin concentrations. It was concluded that amylin does reduce plasma glucagon levels in cats, a feature that has treatment potential in diabetic animals as co administration of amylin would reduce the insulin requirement to control glycaemia. PMID- 19299172 TI - Pharmacokinetic and pharmacodynamic modelling of marbofloxacin administered alone and in combination with tolfenamic acid in goats. AB - In a four-period cross-over study, the fluoroquinolone antibacterial drug marbofloxacin (MB) was administered to goats intramuscularly (IM) at a dose rate of 2 mg/kg, both alone and in combination with the non-steroidal anti inflammatory drug tolfenamic acid (TA), also administered IM at a dose rate of 2 mg/kg. Using a tissue cage model of inflammation, based on the irritant actions of carrageenan, the pharmacokinetics (PK) of MB and MB in combination with TA were determined. MB mean values of area under concentration-time curve (AUC) were similar for serum (5.60 microg h/mL), inflamed tissue cage fluid (exudate; 5.32 microg h/mL) and non-inflamed tissue cage fluid (transudate; 4.82 microg h/mL). Values of mean residence time (MRT) of MB in exudate (15.5 h) and transudate (15.8 h) differed significantly from serum MRT (4.23 h). Co-administration of TA did not affect the PK profile of MB. The pharmacodynamics of MB were investigated using a caprine strain of Mannheimia haemolytica. Integration of PK data with ex vivo bacterial time-kill curve data for serum, exudate and transudate provided AUC(24h)/minimum inhibitory concentration (MIC) ratios of 160, 133 and 121 h, respectively, for the strain of organism used. Modelling of the ex vivo time-kill data to the sigmoid E(max) equation provided AUC(24h)/MIC values required for bacteriostatic and bactericidal actions of MB and for virtual eradication of the organism of 27.6, 96.2 and 147.3 h, respectively. Corresponding values for MB+TA were 20.5, 66.5 and 103.0 h. These data were used to predict once daily dosage schedules of MB for subsequent clinical evaluation. PMID- 19299173 TI - TNT detection with 14N NQR: multipulse sequences and matched filter. AB - Nuclear quadrupole resonance (NQR) has a distinct potential to verify the presence of nitrogen bearing substances based on the unequivocal signatures of their spectra. Therefore, this technique is especially suitable for remote detection of illicit substances and explosives. Unfortunately, the inherent signal-to-noise of the most abundant explosive trinitrotoluene (TNT) is very low. Here we present an NQR method with improved sensitivity for estimation of the probability of TNT presence in the investigated object. The method consists of a spin-lock spin-echo (SLSE) multipulse sequence for signal excitation and a time domain matched filter for signal detection. We find that the signal-to-noise increases by shortening the pulse spacings, even though this means a decrease in spectral resolution. In our case, the decrease of the pulse spacings from the typical 2 ms to 540 micros resulted in an increase of the signal-to-noise by 14 dB. A theory describing this enhancement is presented and compared to experimental results on TNT. Issues related to temperature and polymorphism variations are also discussed. PMID- 19299174 TI - Design of a permanent magnet with a mechanical sweep suitable for variable temperature continuous-wave and pulsed EPR spectroscopy. AB - A magnetic system is introduced which consists of three nested rings of permanent magnets of a Halbach dipolar layout and is capable for EPR spectroscopy. Two of the rings can be rotated independently to adjust the magnetic flux in the center and even allow for mechanical field sweeps. The presented prototype achieves a magnetic flux range of 0.0282-0.3013T with a minimal sweep of 0.15mT and homogeneity of about 10(-3). First applications with CW and pulsed Mims ENDOR as well as ESEEM experiments on a sample of a glycine single crystal doped with 1% copper nitrate demonstrate that flux range, sweep accuracy and homogeneity of this prototype is sufficient for EPR experiments on most solid samples. Together with a recently improved design magnets can be build which could serve as compact and easily transportable replacement of standard electromagnets with negligible consumption of power or coolants. PMID- 19299175 TI - Transaldolase deficiency in two new patients with a relative mild phenotype. AB - Transaldolase (TALDO) deficiency is a recently described inborn error of metabolism of the pentose phosphate pathway that so far has been diagnosed in only eight patients. In this article, we report the clinical course and biochemical findings of two newly identified patients with TALDO deficiency-two sons of consanguineous parents from Polish origin, presenting with neonatal onset of bleeding diathesis, haemolytic anemia, thrombocytopenia and hepatosplenomegaly. Subsequently the patients had persistent thrombocytopenia, a bleeding tendency, impaired liver function and fibrosis. Their physical and psychomotor development progressed normally. PMID- 19299176 TI - Calpain activation is required for homocysteine-mediated hepatic degradation of inhibitor I kappa B alpha. AB - Hepatic steatosis is a clinical feature observed in severe hyperhomocysteinemic patients. In mice, cystathionine beta synthase (CBS) deficiency, the most common cause of severe hyperhomocysteinemia, is also associated with steatosis, fibrosis and inflammation. Proinflammatory cytokines usually induce apoptosis. However, hyperhomocysteinemia does not increase apoptosis in liver of CBS-deficient mice compared to wild type mice. The aim of the study was to analyze the activation state of the NF-kappaB pathway in liver of CBS-deficient mice and to investigate its possible involvement in anti-apoptotic signals. We analyzed the level of I kappaB alpha in liver of CBS-deficient mice. A co-culture of primary hepatocytes and Kupffer cells was also used in order to investigate how I kappaB alpha degradation occurs in response to homocysteine. We found lower I kappaB alpha level not only in liver of CBS-deficient mice but also in hepatocyte/Kupffer cell co-culture. The homocysteine-mediated I kappaB alpha enhanced proteolysis occurred via calcium-dependent calpains, which was supported by an increased level of calpain activity and a reduced expression of calpastatin in liver of CBS deficient mice. Intraperitoneal administration of the inhibitor PDTC normalized the expression of two genes induced by NF-kappaB activation, heme oxygenase-1 and cellular inhibitor of apoptosis 2. Moreover, PDTC administration induced an increase of caspase-3 activity in liver of CBS-deficient mice. Our results suggest that hyperhomocysteinemia induces calpain-mediated I kappaB alpha degradation which is responsible for anti-apoptotic signals in liver. PMID- 19299177 TI - An overview of pulmonary surfactant in the neonate: genetics, metabolism, and the role of surfactant in health and disease. AB - Pulmonary surfactant is a complex mixture of phospholipids (PL) and proteins (SP) that reduce surface tension at the air-liquid interface of the alveolus. It is made up of about 70-80% PL, mainly dipalmitoylphosphatidylcholine (DPPC), 10% SP A, B, C and D, and 10% neutral lipids, mainly cholesterol. Surfactant is synthesized, assembled, transported and secreted into the alveolus where it is degraded and then recycled. Metabolism of surfactant is slower in newborns, especially preterm, than in adults. Defective pulmonary surfactant metabolism results in respiratory distress with attendant morbidity and mortality. This occurs due to accelerated breakdown by oxidation, proteolytic degradation, inhibition or inherited defects of surfactant metabolism. Prenatal corticosteroids, surfactant replacement, whole lung lavage and lung transplantation have yielded results in managing some of these defects. Gene therapy could prove valuable in treating inherited defects of surfactant metabolism. PMID- 19299178 TI - Omni-wedge technique for increased dose homogeneity in head and neck radiotherapy. AB - Dose homogeneity in head and neck tumour irradiation is a challenging task for conventional radiotherapy due to large anatomic contour irregularities and tissue heterogeneities in irradiation volumes. We present a simple technique based on the omni-wedge concept which uses superimposed fields, orthogonally wedged, to improve target-volumes homogeneity in head and neck tumour irradiation. The routine implementation of this technique is straightforward and easy to achieve with standard radiotherapy equipment and treatment planning software. Our five year experience in the clinical implementation of this technique is presented. PMID- 19299179 TI - Severe and long lasting cholestasis after high-dose co-trimoxazole treatment for Pneumocystis pneumonia in HIV-infected patients--a report of two cases. AB - Pneumocystis pneumonia (PCP), a common opportunistic infection in HIV-infected individuals, is generally treated with high doses of co-trimoxazole. However, treatment is often limited by adverse effects. Here, we report two cases of severely immunocompromised HIV-infected patients who developed severe intrahepatic cholestasis, and in one patient lesions mimicking liver abscess formation on radiologic exams, during co-trimoxazole treatment for PCP. Whereas patient 1 showed lesions of up to 1 cm readily detectable on magnetic resonance imaging under prolonged co-trimoxazole treatment, therapy of patient 2 was switched early. PMID- 19299180 TI - [How to analyze an incident of the transfusion chain]. AB - Incidents and accidents analysis is one part of risk management in healthcare organisations. It is based on methods such as healthcare process analysis and the Association of Litigation and Risk Management (ALARM) method, to understand not only immediate causes but also root causes. The goal is to implement relevant remedial and preventive actions to secure healthcare processes. The hospital's risk management unit has to be involved in the methodological analysis of every grade zero transfusion incident. The system is based on a retrospective analysis process in compliance with the French National Authority for Health's ongoing requirements for risk reactive approach, allowing to identify failures and start relevant actions. Healthcare teams should overcome difficulties in order to be totally involved in the process. Human factors recognition, establishment of clear and effective communication between individuals and critical points of control should provide optimum quality care services that are safe and free of unnecessary risks. PMID- 19299181 TI - Impact of short-duration lifestyle intervention in collaboration with general practitioners in patients with the metabolic syndrome. AB - AIM: As the constantly progressing metabolic syndrome is accompanied by an increased risk of type 2 diabetes and cardiovascular complications, it is essential to take appropriate, non-pharmacological, cost-effective measures immediately after the diagnosis has been made. The purpose of our prospective, non-controlled, 6-month study was to determine the impact of lifestyle interventions involving patients' behaviour in collaboration with their general practitioners (GPs). METHODS: We recruited 95 patients (46 men and 49 women, aged 45 to 60 years) who presented with the metabolic syndrome. Each patient received a copy of the national French recommendations (PNNS) leaflet, containing guidelines aimed to balance dietary intake and increase daily physical activity. Socioeconomic status was estimated using the EPICES score. Following a less than 1 hour face-to-face interventional session with each patient to present the lifestyle-modification goals, we contacted each patient's GP by phone to advise on measures to reinforce these lifestyle modifications. RESULTS: The percentage of patients presenting with the metabolic syndrome decreased by 52.4% after 6 months. Hypertension, triglycerides and waist circumference decreased by 30.5, 29.3 and 22.0%, respectively, in the study patients. Rates of compliance to PNNS goals at the last follow-up versus baseline were: for drinks, 63.0% versus 22.2%; for sweet products, 91.4% versus 49.4%; for fat, 91.4% versus 80.3%; and for increased exercise, 26.9% versus 6.4%. CONCLUSION: Short-term, single lifestyle modifications targeting the metabolic syndrome in collaboration with GPs was effective in decreasing most of the parameters of the syndrome. However, no factors predictive of success were identified. PMID- 19299182 TI - Gene polymorphisms and osteoporotic fractures: a study in postmenopausal French women. PMID- 19299183 TI - [OG resident training in France: third stop: CHU of Amiens (Picardie)]. PMID- 19299184 TI - [Required steps to practice Assisted Reproductive Technologies (ART) and role of the French Agence de la biomedicine (ABM)]. PMID- 19299185 TI - [Article reviewing and Evidence-Based Medicine]. PMID- 19299186 TI - [Termination of pregnancy with minors: a feeling of uneasiness]. PMID- 19299187 TI - Toothpaste artifact of the spinal cord observed in a victim of traffic accident who died of pulmonary thrombotic embolism after laminectomy. AB - Toothpaste artifact of the spinal cord is generally known as artificial and morphological change in the field of neuropathology. It is considered that while the spinal cord is pressed on local parts with some solid bodies, the inner spinal tissue pressed penetrates into the weak vicinity of pressed parts and consequently pencil-shaped morphological change, toothpaste artifact, occurs. We report an autopsy case with toothpaste artifact of the spinal cord. A 77-year-old man, who was a front-seat passenger and was injured to the vertebra in a traffic accident, complained suddenly of dyspnea and precordial oppression during a rest on bed in a hospital 18 days after laminectomy, and he died of pulmonary embolism originated in thrombi in the deep veins of lower extremities. The toothpaste artificial change of spinal cord was microscopically observed at the near site of laminectomy. Slight bleeding was observed in a margin of the slipped area, but there was no reactive change. In present case, external compression of spinal cord which occurred accidentally during the period after the operation or at cardiopulmonary resuscitation seems to have caused the toothpaste artifact. PMID- 19299188 TI - Otitis media observed in unexpected natural death of infants. AB - Intracranial complications due to otitis media such as brain abscess and leptomeningitis are well known as a cause of death. In recent years, encountering those serious intracranial complications in forensic medical practice is extremely rare. However, we rarely encounter autopsy cases with otitis media of which pathological damage is mild or moderate. We investigated 11 autopsy cases (6 cases of pneumonia and 5 cases of SIDS) in unexpected natural death of infants under one year old, and found 3 cases with otitis media. The tympanic cavity was investigated by chiseling a petrosal part of the base of the skull. In the case of otitis media, serous and mucous exudate containing leucocytes examined microscopically was observed. Otitis media, as such, was not a cause of death in our cases presented. Background factors causing otitis media seems to be not only functional insufficiency of the auditory tube but also other delicate constitution, hidden dysfunction or immature function in immune system, which could be easily infected. Of 3 cases of otitis media, cytomegalovirus infection was observed in 2 cases simultaneously. In our department, we have little opportunity to encounter autopsy cases of infant under one year old. If many infant cases could be investigated, many autopsy cases with otitis media might be encountered in unexpected infant deaths. PMID- 19299189 TI - Pathomorphological staging of subdural hemorrhages: statistical analysis of posttraumatic histomorphological alterations. AB - We examined 10 histomorphological alterations of 222 cases of subdural hemorrhages following mechanical closed brain injury (MBI) to determine the posttraumatic interval (PTI). These morphological features included red blood cells (RBCs), polymorphonuclear leukocytes (PMNs), macrophages (Ms), RBC containing Ms, hemosiderin-containing macrophages, hematoidin, fibroblasts, endothelial cells, collagenous fibers and membrane formation. The interval between the time of brain injury and death ranged from a few minutes to 33 years. Following routine staining and immunohistochemical staining of macrophages (CD68), paraffin sections were examined by light microscopy for the presence of the selected histomorphological features. An apparent correlation was found between the frequency of a given histomorphological phenomenon and the length of the PTI. Half of the cases (group 1; n=111) were used to develop a multistage evaluation system, the other half (group 2; n=111) to check its accuracy of prediction. Applying this multistage evaluation model and a special software, 85 of the 111 control group cases (76.6%) could be correctly classified and further 21 cases (18.9%) being assigned to an interval close to the correct interval. Therefore, classification of the correct PTI or an interval close to the correct PTI could be achieved in 95.5% of all cases. PMID- 19299190 TI - DNA typing using AmpFlSTR Y-filer for very long-term stored specimens. AB - Aware that long-term storage bloodstain generally reduces numbers of detectable loci, we performed DNA typing with bloodstains stored at room temperature for 22 30years, then extracted DNA from the bloodstains using the AmpFlSTR Y-filer PCR Amplification Kit. We performed electrophoresis with an ABI 310 Genetic Analyzer and determined alleles using GeneMapper ID v3.2 software. Our results suggest that Y-filer finds certain loci more difficult to anneal than others. PMID- 19299191 TI - Legal medicine and the death inquiry system in Japan: a comparative study. AB - In Japan, there are two different systems of death investigation: criminal inspection and judicial autopsy from a criminal justice standpoint and, from a public health standpoint, administrative inspection and either administrative or consent autopsy. As a result, it is sometimes unclear which system is responsible for investigating a death. In this study, the Japanese death inquiry system is compared with the coroner's system used in some other countries. One aim of a death inquiry is to prevent future loss of life in similar circumstances, which the Japanese system does not consider. The systematic reform of the Japanese death inquiry system is thought to be necessary. PMID- 19299192 TI - Urban-rural residence and the occurrence of neural tube defects in Texas, 1999 2003. AB - Neural tube birth defects (NTDs) affect more than 4000 pregnancies in the US annually. The etiology of NTDs is believed to be multifactorial, but much remains unknown. We examined the pattern and magnitude of urban-rural variation in anencephaly, spina bifida without anencephaly, and encephalocele in Texas in relation with urban-rural residence for the period 1999-2003. There was no evidence that urban-rural residence was associated with changes in the rate of anencephaly or spina bifida without anencephaly in unadjusted or adjusted analyses. In contrast, rates of encephalocele were statistically significantly higher in areas classified as suburban or more rural compared to urban areas using four different urban-rural residence indicators. PMID- 19299193 TI - Prenyl transfer to aromatic substrates: genetics and enzymology. AB - Aromatic prenyltransferases catalyze the transfer of prenyl moieties to aromatic acceptor molecules and give rise to an astounding diversity of primary and secondary metabolites in plants, fungi and bacteria. Significant progress has been made in the biochemistry and genetics of this heterogeneous group of enzymes in the past years. After 30 years of extensive research on plant prenylflavonoid biosynthesis, finally the first aromatic prenyltransferases involved in the formation of these compounds have been cloned. In bacteria, investigations of the newly discovered family of ABBA prenyltransferases revealed a novel type of protein fold, the PT barrel. In fungi, a group of closely related indole prenyltransferase was found to carry out aromatic prenylations with different substrate specificity and regiospecificity, and to catalyze both regular and reverse prenylations. PMID- 19299194 TI - Cell kinase activity assay based on surface enhanced Raman spectroscopy. AB - Kinases control many important aspects of cell behavior, such as signal transduction, growth/differentiation, and tumorogenesis. Current methods for assessing kinase activity often require specific antibodies, and/or radioactive labeling. Here we demonstrated a novel detection method to assess kinase activity based on surface enhanced Raman spectroscopy (SERS). Raman signal was obtained after amplification by silver nanoparticles. The sensitivity of this method was comparable to fluorescence measurement of peptide concentration. When purified kinase enzyme was used, the detection limit was comparable to conventional radio labeling method. We further demonstrated the feasibility to measure kinase activity in crude cell lysate. We suggested this SERS-based kinase activity assay could be a new tool for biomedical research and application. PMID- 19299195 TI - Towards automated processing of clinical Finnish: sublanguage analysis and a rule based parser. AB - INTRODUCTION: In this paper, we present steps taken towards more efficient automated processing of clinical Finnish, focusing on daily nursing notes in a Finnish Intensive Care Unit (ICU). First, we analyze ICU Finnish as a sublanguage, identifying its specific features facilitating, for example, the development of a specialized syntactic analyser. The identified features include frequent omission of finite verbs, limitations in allowed syntactic structures, and domain-specific vocabulary. Second, we develop a formal grammar and a parser for ICU Finnish, thus providing better tools for the development of further applications in the clinical domain. METHODS: The grammar is implemented in the LKB system in a typed feature structure formalism. The lexicon is automatically generated based on the output of the FinTWOL morphological analyzer adapted to the clinical domain. As an additional experiment, we study the effect of using Finnish constraint grammar to reduce the size of the lexicon. The parser construction thus makes efficient use of existing resources for Finnish. RESULTS: The grammar currently covers 76.6% of ICU Finnish sentences, producing highly accurate best-parse analyzes with F-score of 91.1%. We find that building a parser for the highly specialized domain sublanguage is not only feasible, but also surprisingly efficient, given an existing morphological analyzer with broad vocabulary coverage. The resulting parser enables a deeper analysis of the text than was previously possible. PMID- 19299196 TI - Hemispheric asymmetry of surround inhibition in the human motor system. AB - OBJECTIVE: Surround inhibition (SI) in the motor system is an essential mechanism for the selective execution of desired movements. To investigate the relationship between the efficiency of SI operation in the motor system and handedness, we performed a transcranial magnetic stimulation (TMS) study in 10 healthy, right handed volunteers. METHODS: TMS was set to be triggered by self-initiated flexion of the index finger at different intervals ranging from 3 to 1000 ms. Average motor evoked potential (MEP) amplitudes obtained from self-triggered TMS were normalized to average MEPs of the control TMS at rest and expressed as a percentage. Normalized MEP amplitudes of the adductor digiti minimi (ADM) and the flexor digitorum superficialis (FDS) muscles were compared between the dominant and non-dominant hands. RESULTS: During index finger flexion, MEP amplitudes of the ADM in the dominant hand were suppressed but not in the non-dominant hand, while MEP amplitudes of the FDS were comparably enhanced in both hands. F-wave amplitudes of ADM were comparably enhanced during index finger flexion in both hands. CONCLUSION: These results suggest that the functional operation of SI in the motor system is more efficient in the dominant hand than the non-dominant hand. More efficient SI in the dominant hand could lead to greater dexterity in the dominant hand. SIGNIFICANCE: Hemispheric asymmetry of SI might be able to serve as a neurophysiological proxy for handedness. PMID- 19299197 TI - Cortical sources of resting-state alpha rhythms are abnormal in persistent vegetative state patients. AB - OBJECTIVE: High power of pre-stimulus cortical alpha rhythms (about 8-12 Hz) underlies conscious perception in normal subjects. Here we tested the hypothesis that these rhythms are abnormal in persistent vegetative state (PVS) patients, who are awake but not aware of self and environment. METHODS: Clinical and resting-state, eyes-closed electroencephalographic (EEG) data were taken from a clinical archive. These data were recorded in 50 PVS subjects (level of cognitive functioning--LCF score: I-II) and in 30 cognitively normal subjects. Rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5 13 Hz), beta 1 (13-20 Hz), and beta 2 (20-30 Hz). Cortical sources were estimated by low-resolution electromagnetic tomography (LORETA). Based on LCF score at 3 months follow-up, PVS patients were retrospectively divided into three groups: 30 subjects who did not recover (NON-REC patients; follow-up LCF: I-II), 8 subjects classified as minimally conscious state patients (MCS patients; follow-up LCF: III-IV), and 12 subjects who recovered (REC patients; follow-up LCF: V-VIII). RESULTS: Occipital source power of alpha 1 and alpha 2 was high in normal subjects, low in REC patients, and practically null in NON-REC patients. A Cox regression analysis showed that the power of alpha source predicted the rate of the follow up recovery, namely the higher its power, the higher the chance to recover consciousness. Furthermore, the MCS patients showed intermediate values of occipital alpha source power between REC and NON-REC patients. CONCLUSIONS: These results suggest that cortical sources of alpha rhythms are related to the chance of recovery at a 3-months follow-up in patients in persistent vegetative state. SIGNIFICANCE: Cortical sources of resting alpha rhythms might predict recovery in PVS patients. PMID- 19299198 TI - The role of altered tissue osmolality on the characteristics and propagation of seizure activity in the intact isolated mouse hippocampus. AB - OBJECTIVES: To study the role of altered tissue osmolality on the characteristics and propagation dynamics of seizure activity and on interictal activity, in a low Mg(+2) artificial cerebrospinal fluid (ACSF) model of recurrent seizures, using the immature (P8-P25) intact isolated mouse hippocampus. METHODS: Recordings were obtained extracellularly from a single site in the CA1 region and from multiple sites along the septotemporal axis measuring spontaneous epileptiform field activity in ACSFs of different osmolalities. RESULTS: In normal osmolar ACSF (310 mOsmol), the average duration of recorded seizures was 90+/-10 s and the average peak amplitude was 0.9+/-0.1 mV. In a hypoosmolar ACSF (270 mOsmol), the seizures were significantly prolonged at 165+/-20 s (p<0.05) with a peak amplitude of 1.2+/-0.3 mV, whereas interictal activity was suppressed. Hyperosmolar ACSF (340 mOsmol) reduced the duration (65+/-15 s) and peak amplitude (0.6+/-0.1 mV, p<0.05) from control, but interictal activity was not affected. No differences in seizure recurrence rate were noted in all three osmolar states. CONCLUSION: The present study, the first to assess of the role of altered tissue osmolality in an intact in vitro preparation, demonstrates that changes in perfusate osmolality play a significant role on the amplitude, duration, and propagation velocity of seizure-like events, and the characteristics of interictal activity, without affecting seizure recurrence rate. SIGNIFICANCE: Increasing tissue osmolality should be considered as a valid target for anticonvulsant treatment. PMID- 19299199 TI - Extrauterine maturation of somatosensory pathways in preterm infants: a somatosensory evoked potential study. AB - OBJECTIVE: To evaluate the reliability of somatosensory evoked potential (SEP) recordings in preterm infants to monitor the intra-uterine and extrauterine maturation of somatosensory pathways. METHODS: We performed SEPs in 35 neurologically normal preterm babies (range 23-35 weeks gestational age--GA). Twenty-four of all infants were evaluated after the first 2 weeks of life, at a minimum post-menstrual age (PMA) of 31 weeks, and 31 at term corrected age. In 15 infants we obtained longitudinal recordings at both epochs. Cross-sectional and longitudinal values of first cortical potential (N1) were analyzed in relation of PMA and matched with those measured in a group of 11 fullterm babies. RESULTS: Reproducible cortical SEPs were found in 92% of preterm babies at first recording, and in all 31 neonates at follow-up. A significant inverse correlation between the latency values of N1 and PMA at the time of first recording was observed, showing that latencies of these components rapidly decrease with increasing PMA. Regression analysis showed no significant effect on N1 latency at term correct age in dependence of GA, suggesting that extrauterine life does not affect maturation of somatosensory pathways. Interestingly, the occurrence of idiopathic respiratory distress (RDS) during clinical course after birth correlated with a delayed N1 latency at term corrected age. CONCLUSIONS: Extrauterine life does not affect maturation of somatosensory pathways in preterms without neurological deficit. Finally, the mild negative influence of RDS on maturational changes was evident. SIGNIFICANCE: SEPs could be considered a useful tool for a non-invasive assessment of somatosensory pathways integrity in preterm infants. PMID- 19299201 TI - The value and limitations of a wound inspection clinic after cardiac device implantation. AB - BACKGROUND: Due to the growing number of cardiac device implantations it is important to develop methods to reduce device-implantation related complications. AIMS: To determine whether a wound inspection clinic can play a role in the detection of device-implantation related complications. METHODS: Single-center observational study evaluating patients who received a pacemaker or implantable cardioverter-defibrillator (ICD). RESULTS: Of 159 patients who received an appointment for the wound inspection clinic, 52 (33%) received a pacemaker and 107 (67%) received an ICD. The majority had no signs of infection. Pain (n = 13,8%) and swelling (n = 11,7%) were the most frequent signs observed, but they never necessitated intervention and recovered spontaneously in all patients. During follow-up (mean 20+/-9 weeks), complications occurred in 10 patients (6%). Most complications occurred early, within 4 days after implantation. The two late complications (at 19 and 41 days) could not be recognized at the wound inspection clinic. CONCLUSION: We found no useful role for a wound inspection clinic two weeks post-implant to detect device-related complications. Open rapid access to the pacemaker/ICD center for patients with signs and symptoms of (threatening) complications seems to be more appropriate to manage post-implant patients. PMID- 19299202 TI - 'It got right to the spot' The patient experience of primary angioplasty: a qualitative study. AB - BACKGROUND: Primary angioplasty is becoming an established treatment for myocardial infarction. Yet we have a limited understanding of patients' attitudes toward this treatment and their experiences of receiving it in practice. AIMS: Exploration of patients' experiences and perceptions of primary angioplasty as treatment for their heart attack. METHODS: Qualitative study of a purposive sample of patients admitted to a tertiary cardiology unit in east London, UK. Participants were interviewed either in their own homes or in hospital. Recruitment continued until no new substantial themes emerged. Eleven men and four women aged 35 to 74 years who had sustained a myocardial infarction were interviewed 13 to 90 days after primary angioplasty. We used the framework method of analysis to identify and explore emerging themes. RESULTS: Informants were expecting open-heart surgery as treatment for their heart attack. They were impressed by the service they actually received. They were generally not engaged in decisions about their treatment in the acute setting and this passivity sometimes persisted after discharge. Expectations of follow up in primary care were not often met. CONCLUSIONS: Patients are satisfied with the primary angioplasty as treatment for acute myocardial infarction but have poor understanding of the management of this condition. PMID- 19299203 TI - Criterion-based clinical audit in obstetrics: bridging the quality gap? AB - The Millennium Development Goal 5 - reducing maternal mortality by 75% - is unlikely to be met globally and for the majority of low-income countries. At this time of heightened concern to scale-up services for mothers and babies, it is crucial that not only shortfalls in the quantity of care - in terms of location and financial access - are addressed, but also the quality. Reductions in maternal and perinatal mortality in the immediate term depend in large part on the timely delivery of effective practices in the management of life-threatening complications. Such practices require a functioning health system - including skilled and motivated providers engaged with the women and communities whom they serve. Assuring the quality of this system, the services and the care that women receive requires many inputs, including effective and efficient monitoring mechanisms. The purpose of this article is to summarise the practical steps involved in applying one such mechanism, criterion-based clinical audit (CBCA), and to highlight recent lessons from its application in developing countries. Like all audit tools, the ultimate worth of CBCA relates to the action it stimulates in the health system and among providers. PMID- 19299204 TI - Conservative management of tubal ectopic pregnancy. AB - In this chapter an overview is given of the best available evidence on the conservative treatment for tubal ectopic pregnancy, i.e., expectant management and medical treatment with systemic methotrexate. From the two randomized controlled trials on expectant management, no conclusions can be drawn yet. It may be that women with low serum hCG levels need not be treated at all, but more research needs to be done in this subgroup of women to reach firm conclusions. Systemic methotrexate in a fixed multiple-dose i/m regimen can be recommended for hemodynamically stable women with an unruptured tubal ectopic pregnancy and no signs of active bleeding presenting with serum hCG concentrations<3,000 IU/l. In women with serum hCG concentrations<1,500 IU/l, a single-dose methotrexate regimen can be considered. PMID- 19299205 TI - Diagnosis and management of ovarian cyst accidents. AB - Ovarian cyst accidents include cyst rupture, haemorrhage and torsion. Torsion commonly occurs to the whole adnexa and is not necessarily associated with an ovarian cyst. Suspected adnexal torsion should always be managed with early laparoscopy and de-torsion of the twisted tube or ovary. Ovarian cyst rupture and haemorrhage usually occur in association with physiological (functional) cysts and are generally self-limiting. Laparoscopy may be necessary in cases where the diagnosis is in doubt or for haemodynamic compromise. Clinical features of ovarian cyst accidents are nonspecific. Ultrasound is the first-line investigation and is diagnostic in the case of haemorrhage. Typical ultrasound findings have been described for ovarian torsion, including an enlarged oedematous ovary with peripheral displacement of follicles. Doppler blood flow findings are variable and not diagnostic. Recurrent cyst rupture or haemorrhage should be prevented by suppression of ovulation, usually with the combined oral contraceptive. Fixation of the ovary by a variety of techniques should be considered to prevent recurrent torsion. PMID- 19299206 TI - Quantitative analysis of biomarkers by LC-MS/MS. PMID- 19299207 TI - Analysis of relationship between cell cycle stage and apoptosis induction in K562 cells by sedimentation field-flow fractionation. AB - Recently, sedimentation field-flow fractionation (SdFFF) was used to study the specific kinetics of diosgenin-induced apoptosis in K562 cells. Here, we propose a new SdFFF cell separation application in the field of cancer research concerning the correlation between induction of a biological event (i.e. apoptosis) and cell status (i.e. cell cycle position). SdFFF isolated subpopulations depending on the cell cycle position allowing the study of apoptosis kinetics and extent. Results showed that cells in G0/G1 phases (F3 cells) underwent significant and earlier apoptosis than cells in the active part of the cell cycle (S/G2/M phases). Results shed light on the correlation between differences in apoptosis kinetics and cell cycle stage when exposure to the inducer began. SdFFF monitoring and size measurement also led to the description of different subpopulations demonstrating complex variations in density between fractions associated with differences in biological processes. PMID- 19299208 TI - Speciation of organotin compounds in urine by GC-MIP-AED and GC-MS after ethylation and liquid-liquid extraction. AB - A method for the determination of organotin compounds in urine samples based on liquid-liquid extraction (LLE) in hexane and gas chromatographic separation was developed and optimized. Seven organotin species, namely monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), tetrabutyltin (TeBT), monophenyltin (MPhT), diphenyltin (DPhT) and triphenyltin (TPhT), were in situ derivatized by sodium tetraethylborate (NaBEt(4)) to form ethylated less polar derivatives directly in the urine matrix. The critical parameters which have a significant effect on the yield of the successive liquid-liquid extraction procedure were examined, by using standard solutions of tetrabutyltin in hexane. The method was optimized for use in direct analysis of undiluted human urine samples and ways to overcome practical problems such as foam formation during extraction, due to various constituents of urine are discussed. After thorough optimization of the extraction procedure, all examined species could be determined after 3 min of simultaneous derivatization and extraction at room temperature and 5 min phase separation by centrifugation. Gas chromatography with a microwave-induced plasma atomic emission detector (MIP-AED) as element specific detector was employed for quantitative measurements, while a quadrupole mass spectrometric detector (MS) was used as molecular specific detector. The detection limits were between 0.42 and 0.67 microg L(-1) (as Sn) for the quantitative LLE-GC-MIP-AED method and the precision between 4.2% and 11.7%, respectively. PMID- 19299209 TI - Using liquid chromatography-tandem mass spectrometry to quantify monohydroxylated metabolites of polycyclic aromatic hydrocarbons in urine. AB - We present an assay which employs enzyme digestion and solid phase extraction followed by liquid chromatography-tandem mass spectrometry to simultaneously quantify 16 hydroxylated polycyclic aromatic hydrocarbons (OHPAHs) in 3-ml samples of urine. The analytes consisted of 2-, 3-, and 4-ring OHPAHs, namely, 1- and 2-hydroxynaphthalene (1- and 2-OHNAP), 2-hydroxyfluorine (2-OHFLU), 1-, 2-, 3 , 4-, and 9-hydroxyphenanthrene (1-, 2-, 3-, 4-, and 9-OHPHE), 1-hydroxypyrene (1 OHPYR), 1- and 2-hydroxybenzo(a)anthracene (1- and 2-OHBAA), 3- and 6 hydroxychrysene (3- and 6-OHCHR) and 3-, 7-, and 9-hydroxybenzo(a)pyrene (3-, 7-, and 9-OHBAP). The method was validated using urine samples from steel workers and control subjects. The coefficients of variation of the method for the particular analytes were between 7% and 27% and the limits of quantitation were between 0.002 and 0.010 microg/l urine. The 2- and 3-ring OHPAHs were easily quantified in all subjects. However, 1-OHPYR was the only representative of the 4- and 5 ring metabolites that could be quantified. Pairwise correlations showed that all OHPAHs were highly correlated with each other (0.553 or = 30) in the promoter of the heme oxygenase-1 (HO-1) gene. Our recent study also demonstrated that far infrared therapy, a noninvasive and convenient therapeutic modality, can improve access flow, inflammatory status and survival of the AVF in HD patients through both its thermal and non-thermal (endothelial-improving, anti-inflammatory, antiproliferative, antioxidative) effects by upregulating NF-E2-related factor-2 dependent HO-1 expression, leading to the inhibition of expression of E-selectin, vascular cell adhesion molecule-1, and intercellular adhesion molecule-1. PMID- 19299218 TI - Bibliometric analysis of obstructive sleep apnea research trends. AB - BACKGROUND: This study aimed to describe current trends and areas of future research using a bibliometric evaluation of the publication output associated with research on obstructive sleep apnea (OSA) during the 16-year period of 1991 2006. METHODS: Data encompassing the period from 1991 to 2006 were extracted from the Science Citation Index online version. We analyzed selected documents with "obstructive sleep apnea", "obstructive apnea", or "OSA" as a part of the title, abstract, or key words and reported the following parameters: trends of publication output, journal pattern, country of publication, authorship, author generated key words, and KeyWords Plus. RESULTS: The annual number of articles on OSA grew at a faster rate than did the number of general scientific publications, from approximately 200 in 1991 to 650 in 2006. The main subject categories in which research on OSA was conducted were the respiratory system and clinical neurology, each of which accounted for > 10% of total articles. Most of the research was conducted in the major industrial countries, with most international collaborations involving the United States and Canada. Certain terms were identified by KeyWords Plus but not by author-generated key words, and some terms increased in frequency of use over time. CONCLUSION: This study provides a bibliometric analysis showing that the annual number of publications related to OSA has been increasing at a much faster rate than the overall scientific literature during the past 16 years in a growing number of specialized journals. Analysis of key words (KeyWords Plus) suggests research trends and areas for future research. PMID- 19299219 TI - Prognostic value of QT parameters in patients with acute hemorrhagic stroke: a prospective evaluation with respect to mortality and post-hospitalization bed confinement. AB - BACKGROUND: This prospective study was performed to evaluate the prognostic prediction value of QT parameters and clinical characteristics exhibited by patients with acute hemorrhagic stroke at the time of presenting to the emergency department (ED). METHODS: One hundred and sixty-six patients admitted to the ED of Taipei Veterans General Hospital from January 2006 to October 2006 because of acute hemorrhagic stroke were enrolled. Glasgow Coma Scale (GCS) scores between 3 and 8 were taken to indicate severe neurologic deficits. QT parameters (QT max, QT min, QT dispersion, QTc max, QTc min, QTc dispersion) and other pertinent clinical variables were determined on admission. Logistic regression model was applied to evaluate prognostic prediction values. RESULTS: Mortality was higher among stroke patients with low GCS scores (p < 0.01). Leukocyte counts and systolic blood pressures were significantly higher among non-surviving patients (p = 0.04). No association was found between QT parameters and mortality (all p > 0.05). Among survivors, post-hospitalization bed confinement was required for those significantly older (p = 0.01) and those with higher QT max and QTc max values in multivariate analyses (p = 0.04 and p < 0.01, respectively). CONCLUSION: Low GCS scores, increased leukocyte counts, and elevated systolic blood pressures predict increased mortality for subjects with acute hemorrhagic stroke. Advanced age and prolongations in QTc and QT max at the time of stroke predicted poor functional recovery for these subjects. PMID- 19299220 TI - Relationship between central corneal thickness, refractive error, corneal curvature, anterior chamber depth and axial length. AB - BACKGROUND: To determine the relationship between central corneal thickness (CCT), refractive error, corneal curvature, anterior chamber depth and axial length in normal Taiwanese Chinese adults. METHODS: Five hundred normal Taiwanese Chinese patients aged 40-80 years were recruited for the study. Measurement procedures included CCT, refractive error, corneal curvature, anterior chamber depth and axial length. Exclusion criteria were previous ocular surgery, glaucoma, trauma history, external eye disease, and previous contact lens use. The relationships among parameters were tested using Pearson's correlation and linear regression analysis. RESULTS: The median CCT was 555 +/- 27 mum for males and 553 +/- 30 mum for females. Eyes with more myopic refractive error tended to have greater axial length (r = -0.645, p < 0.001). Eyes with axial elongation tended to have flatter cornea (r = -0.502, p < 0.001) and deeper anterior chamber (r = 0.651, p < 0.001). There were no significant correlations between the CCT and refractive error (r = -0.034, p = 0.445), corneal curvature (r = 0.013, p = 0.770), anterior chamber depth (r = 0.023, p = 0.614) and axial length (r = 0.053, p = 0.223). CONCLUSION: CCT was not associated with refractive error, corneal curvature, anterior chamber depth and axial length. CCT is an independent factor unrelated to other ocular parameters. PMID- 19299221 TI - A facile technique to make articulating spacers for infected total knee arthroplasty. AB - BACKGROUND: To treat total knee arthroplasty, 2-stage revision, including removal and reimplantation, remains the standard treatment for the infected arthroplasty. Articulating cement spacer has been shown to provide better functional results after reimplantation. However, its cost as a manufactured product is not always easily affordable and the choice of antibiotics is not flexible either. The authors have developed a method for surgeons to make cement-on-cement articulating spacers themselves by using an impression-taking technique with polydimethyl siloxane. The current study was conducted to test their clinical efficacy. METHODS: Fifteen patients with infected total knee arthroplasties were prospectively treated with 2-stage revision using articulating spacers made by this technique. The clinical assessment included intraoperative finding, surgical records, radiographic and laboratory examination and final functional scores. All the patients were regularly followed-up. RESULTS: Fourteen of the 15 patients (93.3%) had infection eradicated, of which 13 patients received revision arthroplasty successfully. The average interval between the resection arthroplasty and the final procedure was 3.5 months. During this period, most of the patients could sit comfortably with bent knees and walk with partial weight bearing. No patients had secondary bone loss. The range of motion after revision surgery achieved an average of 110 degrees. The average Hospital for Special Surgery score was 90.5 points, and none had recurrent infection after an average of 47.5 months of follow-up. CONCLUSION: Treating infected total knee arthroplasty with these self-made articulating spacers eradicates infection effectively, improves the life quality before reimplantation and provides good final results without significant complications. PMID- 19299222 TI - Acute embolic myocardial infarction in a patient with paroxysmal atrial fibrillation receiving direct-current cardioversion. AB - Coronary embolism with acute myocardial infarction (MI) following direct-current (DC) cardioversion of atrial fibrillation (AF) has rarely been reported. We present the case of a 34-year-old female with severe aortic regurgitation and highly symptomatic paroxysmal AF. Acute embolic MI occurred 4 days after DC cardioversion of AF, although there was no left atrial thrombus detected before this procedure. Insufficient anticoagulation therapy during the post cardioversion period was the cause, leading to embolic MI. PMID- 19299223 TI - Nerve invasion by epithelial cells in benign breast diseases. AB - Nerve invasion by glandular epithelial cells in a lesion is usually regarded as invasive carcinoma. However, some benign conditions in the pancreas, prostate, breast and other organs may show involvement of nerve bundles by benign epithelial cells. We report an 18-year-old female with nerve invasion in benign breast disease. The lesion in her right breast revealed fibrocystic changes with ductal hyperplasia and stromal sclerosis. Perineural and intraneural involvement by bland-looking small ducts lined by 2 layers of cells including an outer layer of myoepithelial cells were found, suggestive of benign nerve invasion. There was no evidence of malignant cells in any of the sections. The patient remains well after 31 months of follow-up. About 44 cases of nerve invasion in benign breast diseases have been reported in the literature. It is necessary to carefully evaluate nerve involvement in breast lesions to avoid over-diagnosis and inappropriate operation. PMID- 19299224 TI - Recurrent retroperitoneal abscess due to perforated colonic diverticulitis in a patient with polycystic kidney disease. AB - Patients with autosomal dominant polycystic kidney disease (ADPKD) usually have extrarenal manifestations. We report the case of a 53-year-old man with ADPKD who presented with a retroperitoneal abscess of the left side after undergoing bilateral nephrectomy for a cyst that had repeatedly bled for 1 year. The abscess recurred despite drainage with a pig-tail catheter and antibiotic treatment. Fistulography with injection of diluted contrast medium via the pig-tail catheter showed an accumulation of contrast medium in the descending colon, which indicated a fistula between the abscess and the descending colon. A portion of the descending colon was resected, and multiple diverticulitis with 1 perforation in the resected specimen was observed. The findings support a diagnosis of retroperitoneal abscess caused by a perforated diverticulum---an extrarenal manifestation of ADPKD. PMID- 19299225 TI - Pregnancy-induced hemophagocytic lymphohistiocytosis combined with autoimmune hemolytic anemia. AB - Hemophagocytic lymphohistiocytosis (HLH), presenting with fever, cytopenia, liver dysfunction, hepatosplenomegaly, hypertriglyceridemia, and hyperferritinemia, is associated with various etiologies, including infections, collagen vascular diseases, and malignancies. The present report describes a 28-year-old woman who developed HLH combined with autoimmune hemolytic anemia (AIHA) at 23 weeks of gestation. Without response to corticosteroid, the patient completely recovered from both HLH and AIHA after termination of the pregnancy. Pregnancy-induced immune dysregulation and cytokine overproduction in genetically susceptible women may play critical roles in HLH. The differential diagnosis of pregnant women with fever and cytopenia should include HLH. Pregnancy termination should be considered when pregnancy-induced HLH is refractory to medical treatment. PMID- 19299226 TI - Emphysematous pyelonephritis presenting as necrotizing fasciitis of the leg. AB - We report a 50-year-old man with poorly controlled diabetes mellitus who presented with a painful, swollen right leg. He had also experienced right flank pain for 1 week prior to admission. Physical examination was notable for tenderness over the right flank. The right leg was diffusely swollen and exquisitely tender to touch, with palpable crepitance. Laboratory tests revealed leukocytosis and pyuria. Computed tomography showed a right ureteral stone with hydronephrosis and characteristic findings of emphysematous pyelonephritis. Furthermore, a right perirenal gas-forming abscess with extension to the right leg was noted. The patient was successfully treated with antibiotic therapy, aggressive control of blood sugar, percutaneous drainage of the hydronephrosis and perirenal abscess, and aggressive debridement of the leg. PMID- 19299227 TI - Bilateral diaphragmatic defect and associated multiple anomalies. AB - Although congenital diaphragmatic hernia is one of the most common congenital anomalies, complete bilateral agenesis of the diaphragm is a very rare congenital malformation and frequently associated with other major anomalies. We report a case of bilateral diaphragmatic agenesis associated with major congenital anomalies. A 2,240-g male infant was born at 35 weeks of gestation to a 34-year old mother with a history of minimal prenatal care. Polyhydramnios was reported on prenatal level 1 scan. The patient experienced early respiratory distress requiring intubation. Apgar scores were 2/1/1 at 1, 5 and 20 minutes, respectively, and efforts to resuscitate him were unsuccessful. He died at 2 hours of age. Autopsy revealed bilateral diaphragmatic agenesis associated with right pulmonary hypoplasia, left pulmonary agenesis, multiple cardiac abnormalities and gallbladder agenesis. Cytogenetic studies showed normal male karyotype. Bilateral agenesis of the diaphragm is a life-threatening malformation. Survival of these infants often depends on cardiopulmonary function. Bilateral agenesis of the diaphragm associated with gallbladder and unilateral pulmonary agenesis is a rare entity, and its clinical significance needs further investigation. PMID- 19299228 TI - The first Asian case report of eosinophilic esophagitis in an asymptomatic adult: what about a proton pump inhibitor trial? PMID- 19299230 TI - Atypical glycine encephalopathy in an extremely low birth weight infant: description of a new mutation and clinical and electroencephalographic analysis. AB - We present the clinical course and EEG evolution of an extreme low birth weight preterm neonate with an uncommon type of glycine encephalopathy. The patient presented with myoclonic jerks, apnea and encephalopathy three months after birth without satisfactory therapeutic response. During the first days of clinical symptoms the patient presented a paroxystic burst-attenuation EEG pattern which progressively evolved into an established typical burst-suppression pattern within a few days. West syndrome occurred four weeks later and the patient died at seven months of extra-uterine life due to a serious respiratory infection with cardio-respiratory arrest. Genetic analysis showed a non-previously described mutation affecting a consensus splice site (IVS2-1G > C 3) in the AMT gene encoding the T protein of the glycine cleavage system. PMID- 19299231 TI - Mirthful gelastic seizures with ictal involvement of temporobasal regions. AB - Ictal laughter is the cardinal clinical sign of gelastic seizures in hypothalamic hamartomas and may also occur in extrahypothalamic epilepsies. Laughing consists of an affective and a motor component. It has been suggested that the affective component may result from an involvement of temporobasal structures, whereas the motor part is related to an involvement of the mesial frontal cortex. So far, evidence is based on a limited number of cases with spontaneously recorded seizures or in whom electrical stimulation of invasive intracranial EEG recordings has been performed. We report a patient who suffered from epigastric psychic auras, complex partial seizures with a gelastic component and secondarily generalized seizures. To evaluate a possible epileptogenic role of the hippocampus and dysplastic region in the right mid-temporal gyrus, intracranial monitoring with subdural electrodes over the temporobasal and temporolateral regions, as well as a deep brain electrode in the hippocampus, were performed. During the intial part of the seizure, consisting of an intense retrosternal ascending feeling with sexual connotation, rhythmic spikes in temporolateral contacts were detected. Concomitant with the development of smiling and laughter, a rhythmic activity over the temporobasal regions evolved. The patient became seizure-free following right temporal lobe resection. This case supports the assumption that ictal involvement of temporobasal structures is crucial for gelastic seizure components in patients with temporal lobe epilepsy. Progression to temporobasal regions was associated with the feeling of happiness whereas motor components of laughter occurred later. These findings are in accordance with the interpretation of surface recordings by Dericioglu and co-workers in a similar previous case. [Published with video sequences]. PMID- 19299232 TI - Clinical breast cancer. PMID- 19299233 TI - Innumerable variations: combining biologics. PMID- 19299235 TI - A phase I/II trial of trastuzumab plus erlotinib in metastatic HER2-positive breast cancer: a dual ErbB targeted approach. AB - BACKGROUND: This phase I/II trial was conducted to determine the toxicities, recommended dose, pharmacokinetics, and response rate of erlotinib plus trastuzumab in metastatic HER2+ breast cancer. PATIENTS AND METHODS: In phase I, sequential groups of patients with unlimited previous treatment received erlotinib at dose levels of 50, 100, and 150 mg plus standard dose weekly trastuzumab. In phase II, only patients with no previous chemotherapy or trastuzumab in the metastatic setting were allowed. RESULTS: The combination was well tolerated among the 16 patients enrolled in phase I, and the recommended phase II dose of erlotinib was initially set at 150 mg. After an interim review of the first 8 patients in phase II revealed a higher incidence of rash and diarrhea than expected from the phase I experience, the protocol was amended to treat new phase II patients at erlotinib 100 mg, with the opportunity to escalate to 150 mg after 3 weeks, based on individual patient tolerability. As a result of advances in other therapies aimed at HER2+ breast cancer, phase II closed before meeting its accrual goal. Among the 12 evaluable chemotherapy- and trastuzumab naive patients treated at the recommended phase II dose level, there were 4 partial responses, and the time to progression was 9.03 months (95% CI, 1.2 undetermined). No pharmacokinetic interaction between the 2 agents was observed. CONCLUSION: The combination of erlotinib and trastuzumab was well tolerated when the dose of erlotinib was tailored to individual patient experience, and there was preliminary evidence of anticancer activity. PMID- 19299236 TI - Phase II trial of weekly docetaxel, vinorelbine, and trastuzumab in the first line treatment of patients with HER2-positive metastatic breast cancer. AB - BACKGROUND: The combinations of trastuzumab/docetaxel and trastuzumab/vinorelbine are highly active in the treatment of patients with HER2-positive metastatic breast cancer (MBC). We investigated the feasibility and safety of a 3-drug combination of trastuzumab, docetaxel, and vinorelbine as first-line therapy in this patient group. PATIENTS AND METHODS: Sixty patients with previously untreated, measurable HER2-positive MBC (immunohistochemistry 3+ and/or fluorescence in situ hybridization positive) were treated with docetaxel 30 mg/m2 intravenously (I.V.) and vinorelbine 25 mg/m2 I.V. on days 1 and 8 of each 3-week cycle. Trastuzumab was given weekly (4-mg/kg loading dose followed by 2 mg/kg/week). Patients were evaluated after 6 weeks; responders/stable patients continued treatment until progression. RESULTS: Patients received a median of 11 treatment cycles (range, 1-22 cycles). Forty-one of 60 patients (68%) had major responses (16 complete responses [27%], 25 partial responses [42%]). An additional 13 patients (22%) had stable disease for > or = 6 months. After a median follow-up of 58 months, median progression-free survival was 12 months (95% CI, 9.1-16.3 months), and the median overall survival was 40.8 months (95% CI, 25-not reached). Neutropenia (72% grade 4) was the most common hematologic toxicity; 8 patients were hospitalized for febrile neutropenia. A total of 67% of patients required dose modifications for neutropenia during cycles 1 or 2. Other grade 3/4 toxicities included fatigue (12%), hyperglycemia (7%), and myalgias (7%). There were no treatment-related deaths. CONCLUSION: The combination of trastuzumab, docetaxel, and vinorelbine is highly active as first-line treatment for patients with HER2-positive MBC. However, this regimen offers no obvious advantages over other less myelosuppressive trastuzumab-containing regimens, and its routine use is not supported by the study. PMID- 19299237 TI - Survival outcomes for patients with metastatic triple-negative breast cancer: implications for clinical practice and trial design. AB - BACKGROUND: Clinical experience suggests that many women with triple-negative metastatic breast cancer (MBC) relapse quickly. This has implications for clinical practice and trial design. We evaluated the duration of first-, second-, and third-line chemotherapy as a surrogate for duration of treatment response. PATIENTS AND METHODS: We performed a retrospective multicenter chart review of patients with triple-negative MBC receiving palliative chemotherapy. Primary outcome was duration of palliative chemotherapy, and secondary outcome was to identify prognostic variables. RESULTS: A total of 111 patients were analyzed. Median age at diagnosis was 51 years (range, 26-82 years). Fourteen percent of patients presented with MBC. Twenty-seven percent received neoadjuvant chemotherapy, and 48% received adjuvant chemotherapy. Median distant disease-free interval (DDFI) was 18 months (range, 0-172 months). At presentation of MBC, 68% had visceral and 71% had multiple sites of disease. Median survival with MBC was 13.3 months (range, 0.8-99.8 months). Median duration of first-line palliative therapy was 11.9 weeks (range, 0-73.1 weeks). Eighty-seven patients (78%) went on to receive second-line therapy with a median duration of 9 weeks (range, 0-120.9 weeks), and 55 (49%) received third-line therapy with a median duration of 4 weeks (range, 0-59 weeks). Multivariate analysis revealed that age < 50 years, ALP > 120 U/L, history of previous chemotherapy, DDFI < 12 months, and visceral presentation were all independently associated with a poor prognosis. CONCLUSION: Despite the poorer overall prognosis of patients with triple-negative disease, there remains considerable heterogeneity in individual outcomes. Many women with recurrent triple-negative disease will progress quickly on first-, second-, and third-line palliative treatment. Future clinical trials in this population must take into account their shorter time to progression when determining optimal trial design. PMID- 19299238 TI - Aromatase inhibitor-related musculoskeletal symptoms: is preventing osteoporosis the key to eliminating these symptoms? AB - BACKGROUND: Aromatase inhibitors (AIs) are an effective treatment for postmenopausal women with hormone receptor-positive breast cancer. However, patients receiving AIs report a higher incidence of musculoskeletal symptoms and bone fractures; the mechanism and risk factors for this correlation are not well studied. The aim of this study was to correlate these musculoskeletal symptoms and bone fractures in patients receiving AIs with bone mineral density (BMD), previous tamoxifen use, and administration of calcium/bisphosphonate (Ca/Bis). PATIENTS AND METHODS: We reviewed charts of 856 patients with hormone receptor positive nonmetastatic breast cancer seen at our institution between January 1999 and October 2007. A total of 316 patients met the inclusion criteria of treatment with one of the AIs for > or = 3 months and availability of a dualenergy X-ray absorptiometry (DEXA) during this treatment. Arthralgia, generalized bone pain and/or myalgia, bone fracture after beginning AIs, any tamoxifen treatment, and Ca/Bis therapy were recorded. RESULTS: Our study demonstrates a significant association between symptoms and DEXA-BMD results (P < .001). Similarly, the group receiving tamoxifen before AIs had fewer patients with arthralgia or generalized bone pain/myalgia or bone fracture (P < .001). Furthermore, the group receiving AIs plus Ca/Bis had more patients without musculoskeletal symptoms and had fewer fractures. Finally, the group receiving steroidal AIs compared with nonsteroidal AIs had more patients with arthralgia or generalized bone pain and/or myalgia, and bone fractures (P < .001). CONCLUSION: Patients on AIs who develop osteoporosis are at increased risk of musculoskeletal symptoms and bone fracture. Comedication with Ca/Bis reduces the likelihood for osteoporosis and musculoskeletal symptoms. Patients who received tamoxifen before AIs were less likely to develop AI-related musculoskeletal symptoms. We recommend that patients on AIs should be offered Ca/Bis to reduce the incidence of musculoskeletal symptoms and fracture, especially if patients are receiving steroidal AI and/or did not receive tamoxifen before AIs. PMID- 19299239 TI - A comparative study of exemestane versus anastrozole in patients with postmenopausal breast cancer with visceral metastases. AB - PURPOSE: Patients developing visceral breast cancer metastases generally receive chemotherapy rather than endocrine therapy. Recent aromatase inhibitor studies have reported activity in such patients; therefore, this study formally evaluated anastrozole and exemestane in postmenopausal patients in this setting. PATIENTS AND METHODS: Postmenopausal women with advanced breast cancer and > or = 1 visceral (liver or lung) lesion were randomized to anastrozole (1 mg/day orally) or exemestane (25 mg/day orally) for > or = 8 weeks. The primary endpoint was objective response in visceral lesions based on modified Response Evaluation Criteria in Solid Tumors. Secondary endpoints included clinical benefit (objective response plus stable disease > or = 180 days), overall survival, and adverse events. RESULTS: A total of 130 patients were enrolled, and 128 patients (64 anastrozole, 64 exemestane) were included in the intent-to-treat analysis. Accrual delays caused study closure before the target enrollment (N = 200) was reached, limiting the statistical power of the study. Objective response in visceral sites was approximately 15% in both groups. Clinical benefit in visceral sites was 32% of the patients treated with anastrozole and 38% of the patients treated with exemestane. Median survival was 33.3 months and 30.5 months in the anastrozole and exemestane groups, respectively. Toxicities were similar to those previously reported; however, treatment-related adverse events were more frequent with anastrozole (41%) than with exemestane (31%). Both treatments were generally well tolerated in patients with postmenopausal breast cancer with visceral metastases. CONCLUSION: Efficacy was similar in both treatment groups for all endpoints. Aromatase inhibitors can be considered as a treatment option in postmenopausal patients with hormone receptor-positive visceral breast cancer metastases. PMID- 19299240 TI - Incidence of invasive breast cancer in postmenopausal women after discontinuation of long-term raloxifene administration. AB - BACKGROUND: Postmenopausal women with osteoporosis had a 66% relative risk reduction for invasive breast cancer over 8 years of raloxifene therapy in the randomized, placebo-controlled 4-year MORE (Multiple Outcomes of Raloxifene Evaluation) trial and the CORE (Continuing Outcomes Relevant to Evista) trial, a 4-year follow-up to MORE. PATIENTS AND METHODS: The first post hoc analysis examined the effects of raloxifene on the cumulative incidence of invasive breast cancer on a yearly basis. Another analysis compared the incidence of invasive breast cancer in 3967 patients who continued raloxifene for 8 years (RLX-C, n = 2280), discontinued raloxifene after 4 years in MORE (RLX-D, n = 401), or took placebo (n = 1286) for a mean 2.9 years' treatment duration (57,338 patient years). RESULTS: The unadjusted breast cancer incidence rate was 5.39 per 1000 patient-years in the placebo group compared with 2.26 in the RLX-C group (hazard ratio [HR], 0.41 [95% CI 0.21-0.81]) and 3.59 in the RLX-D group (HR, 0.69 [95% CI 0.23-2.01]). Because the choice of taking the study drug was not randomized in CORE, propensity scores were used to adjust for potential imbalances in baseline characteristics before CORE. Results after adjustment by the propensity score method were similar to the unadjusted results. CONCLUSION: This analysis suggests a persistent effect for breast cancer risk reduction in patients who discontinued raloxifene, although this conclusion is limited by the small sample size. PMID- 19299241 TI - Hepatitis C virus infection does not preclude standard breast cancer-directed therapy. AB - In the United States, over 3 million people are infected with hepatitis C virus (HCV), and over 200,000 women develop breast cancer annually. Yet, no published studies have investigated the tolerability of breast cancer- directed therapy among women with HCV infection. We reviewed records at Memorial Sloan-Kettering Cancer Center and identified 35 patients with chronic HCV infection who were treated for breast cancer between the years 1991 and 2005. One (2.9%) of 35 also had chronic hepatitis B virus infection. There were no complications related to HCV infection during or after surgery or radiation therapy. A total of 29 (82.9%) of the 35 patients received chemotherapy for breast cancer. Of the 29, only 4 required chemotherapy delays or adjustments in dosing because of HCV infection. In conclusion, breast cancer therapy was well tolerated among women with HCV infection. Considering the paucity of complications, routine screening for HCV infection is not warranted among women with breast cancer and no defined risk factors for HCV infection. PMID- 19299242 TI - Bowen's disease of the nipple in a young man with AIDS: a case report. AB - Bowen's disease, or squamous cell carcinoma in situ (SCCIS) of the skin, is a malignant neoplasm restricted to the epidermis, without evidence of dermal invasion. It usually develops in sun-exposed area of skin, but other sites can also be affected. Bowen's disease of the nipple is extremely rare and has thus far been reported only in women. We present the case of Bowen's disease of the nipple in an HIV-positive male patient who presented with a scaly lesion on nipple for one year. He also had past genital infection with human papillomavirus, but he was found to be negative for high-risk subtypes. Biopsy of the lesion revealed SCCIS of the nipple areola complex, with extension into the underlying lactiferous ducts of the breast. There was no evidence of invasive carcinoma. The patient was treated with a simple mastectomy with sentinel lymph node biopsy. With the advent of highly active antiretroviral treatment, chronic non-HIV related conditions have become more important, male breast cancer being one of them. To the best of our knowledge, this is the first reported case in the worldwide literature of Bowen's disease of the nipple in a young immunocompromised male patient. More aggressive therapy in HIV-positive male patients presenting with precancerous and cancerous breast lesions is recommended. PMID- 19299243 TI - Rational and successful use of carboplatin and albumin-bound paclitaxel in a patient with recurrent metaplastic carcinoma who presented with multi-organ tumor emboli. AB - We demonstrate successful treatment of recurrent chondroid-metaplastic breast cancer. Breast cancer recurrence was diagnosed when a patient with a history of metaplastic breast cancer presented with recurrent acute strokes. A diagnosis of tumor embolism was suspected when a chest radiograph performed as part of a work up for stroke demonstrated several lung nodules, with 1 lung nodule invading the pulmonary vein and extending into the left atrium-the source of tumor emboli. This was followed by timely surgery to remove the embolizing metastatic lesion and local radiation to prevent growth and recurrent embolization. Subsequently, the patient received carboplatin and albumin-bound paclitaxel and experienced complete remission. PMID- 19299245 TI - Matrix metalloproteinases and tissue inhibitors of metalloproteinases in gingival crevicular fluid during orthodontic tooth movement. AB - Orthodontic tooth movement requires extensive re-modelling of the periodontium. Matrix metalloproteinases (MMPs) degrade the extracellular matrix during re modelling, while their activity is regulated by the tissue inhibitors of metalloproteinases (TIMPs). The aim of this study was to investigate differences in MMP and TIMP levels in the gingival crevicular fluid (GCF) at the resorption and apposition sides of orthodontically moved teeth, and to compare these with control teeth. GCF samples were collected from eight orthodontic patients wearing fixed appliances with superelastic nickel-titanium coil springs. The samples were analysed by gelatin zymography, which allows detection of both active and latent MMPs, and reverse zymography for analysis of TIMPs. Western blotting was performed to confirm the identity of MMPs. The data were analysed using either the one-way analysis of variance or the Kruskal-Wallis test. In general, higher levels of MMPs and TIMPs were found at both the resorption and apposition sides compared with the control teeth. Remarkably, partially active MMP-1 was found in GCF from both the resorption and the apposition side but was barely present at the control teeth. TIMP-1 was strongly increased at the apposition side. Gelatinases were mainly present at the resorption side, while gelatinolytic fragments were exclusively detected at the apposition side. MMP-9, which is known to be involved in bone degradation, and a 48 kDa gelatinase were increased at the resorption side. The small increase in TIMP-1 at the resorption side might stimulate bone resorption, whereas the large increase at the apposition side reduces bone resorption. The analysis of MMPs and TIMPs may contribute to the improvement of orthodontic treatment regimens. PMID- 19299246 TI - Microcytic anaemia can 'mask' co-existing cobalamin deficiency. PMID- 19299247 TI - Severity assessment in community-acquired pneumonia: a review. AB - Severity assessment is an important early step in the management of patients presenting with community-acquired pneumonia. Various pneumonia-specific scores, generic sepsis scores and predictive biomarkers have been proposed as tools to aid clinicians in key management decisions. However, there is no uniform agreement about the optimum severity assessment tool to use. This review provides a summary of current evidence surrounding severity assessment in adult patients presenting with community-acquired pneumonia. PMID- 19299248 TI - History of the cholesterol hypothesis in Britain. PMID- 19299249 TI - Voodoo dolls and the cancer patient: patients do trust their doctors. AB - OBJECTIVES: To determine oncology patients' pattern and rationale of complementary and alternative medicine (CAM) use, and canvass their views on the relative merits of allopathic and alternative medicine. DESIGN: Observational study of opinions from a cohort of patients using self-completion questionnaires. SETTING: Oncology departments of two UK teaching hospitals. PARTICIPANTS: Voluntary participation of 200 oncology patients attending clinic. MAIN FINDINGS: Twenty-two percent of patients used CAM, with a preponderance towards younger, female patients. The commonest reasons for CAM use is to make the patient feel better and to help with their cancer. However, patients seldom believe there is more evidence for CAM or that CAM will cure them, indeed often noticing no benefits from the treatment. CAM users do not resort to complementary medicine due to dissatisfaction with their doctor but instead have considerable trust in their physicians. Only a minority believes their doctor knows about their CAM use. CONCLUSION: CAM use by oncology patients in the UK is less common than that reported elsewhere. Although patients try CAM in the hope that it will help with their treatment, they are realistic about its likely benefits. It uptake is not as an indication of lack of faith in doctors, yet physicians are frequently unaware of use. Therefore, the medical profession should not feel threatened by patients resorting to CAM but instead focus on understanding the reasons behind it. PMID- 19299250 TI - The perception of elder sexual abuse in the courtroom. AB - This study explored mock juror perceptions of elder sexual mistreatment (ESM). In Experiment 1, 118 participants read a fictional criminal trial summary of an ESM case in which a 76-year-old woman was allegedly abused by either her son or a neighbor. In Experiment 2 (n = 360), the ESM occurred in either a nursing home or the elder's home and the alleged perpetrator was either her son or a nursing home worker. Conviction rates were relatively low in both experiments (25% and 33%, respectively). Women were more pro-prosecution than men in case judgments. Overall, the study provides evidence that mock jurors may question the credibility of elders in ESM cases. PMID- 19299251 TI - Trophoblast stem cells: models for investigating trophectoderm differentiation and placental development. AB - The placenta is an ephemeral organ containing diverse populations of trophoblasts that are all derived from the embryonic trophectoderm but have morphological, functional, and molecular diversity within and across species. In hemochorial placentation, these cells play especially important roles, interfacing with and modifying the cells of the maternal decidua. Within the rapidly growing placenta, it has been shown that there are trophoblast stem cells well characterized in the mouse and postulated but not well understood in primates. This review will discuss the characteristics of candidates for human and nonhuman primate trophoblast stem cells, present the diverse methods of their generation, and propose future prospects for experimental systems in which they can shed light on developmental and pathophysiological processes in human pregnancy. PMID- 19299252 TI - Alagebrium chloride protects the heart against oxidative stress in aging rats. AB - To investigate the possible effects of alagebrium chloride (ALT-711) on oxidative stress (OS) process in aging hearts, we examined the role of ALT-711 in cardiac function and OS in the heart of aging rats. Increased mitochondrial DNA (mtDNA) deletion as well as nearly a twofold increase in advanced glycation end products (AGEs) accumulation were observed in aging heart, whereas only about 50% of the superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities were seen. However, after treatment with ALT-711, preserved cardiac diastolic function accompanied with reduced mtDNA deletion and about 30% of AGEs decrease was observed in aging hearts. In addition, ALT-711 can increase SOD and GSH-PX activities in aging hearts as well as in cultured cardiomyocytes. In conclusion, our study suggests that AGEs accumulation and the abnormalities in the OS in aging hearts can be attenuated by ALT-711, and this might be a novel underlying mechanism for ALT-711 in the treatment of cardiovascular diseases that develop with aging. PMID- 19299253 TI - Distinct genetic influences on cortical surface area and cortical thickness. AB - Neuroimaging studies examining the effects of aging and neuropsychiatric disorders on the cerebral cortex have largely been based on measures of cortical volume. Given that cortical volume is a product of thickness and surface area, it is plausible that measures of volume capture at least 2 distinct sets of genetic influences. The present study aims to examine the genetic relationships between measures of cortical surface area and thickness. Participants were men in the Vietnam Era Twin Study of Aging (110 monozygotic pairs and 92 dizygotic pairs). Mean age was 55.8 years (range: 51-59). Bivariate twin analyses were utilized in order to estimate the heritability of cortical surface area and thickness, as well as their degree of genetic overlap. Total cortical surface area and average cortical thickness were both highly heritable (0.89 and 0.81, respectively) but were essentially unrelated genetically (genetic correlation = 0.08). This pattern was similar at the lobar and regional levels of analysis. These results demonstrate that cortical volume measures combine at least 2 distinct sources of genetic influences. We conclude that using volume in a genetically informative study, or as an endophenotype for a disorder, may confound the underlying genetic architecture of brain structure. PMID- 19299254 TI - Attitudes of adolescent Spanish Roma toward noninjection drug use and risky sexual behavior. AB - Our objective with this study was to analyze the opinions of a potentially socially marginalized group of Spanish Roma adolescents and young adults about noninjection drug use, HIV infection, and risky sexual behavior. Descriptive qualitative research was conducted through focus groups with Roma participants and semistructured interviews with professionals who work with them in areas such as education and health promotion. Results were triangulated by cross analysis among researchers. Declared drug consumption is lower among females than males. The former claim they do not maintain sporadic sexual relations, and link risky sexual practices to being in love and involved in a stable relationship. Males only use condoms in sporadic sexual relations. They attribute their lack of condom use to lost sensitivity, perceiving sex as something uncontrollable, and not having condoms available when using drugs. Results suggest the need to improve actions aimed at preventing the sexual transmission of HIV among the Roma population. PMID- 19299255 TI - The role of contingency and contiguity in young and older adults' causal learning. AB - Contingency and temporal contiguity are important "cues to causality." In this study, we examined how aging influences the use of this information in response outcome causal learning. Young and older adults judged a generative causal contingency (i.e., outcome is more likely when a response is made) to be stronger when response and outcome were contiguous than when the outcome was delayed. Contiguity had a similar beneficial effect on young adults' preventative causal learning (i.e., outcome is less likely when a response is made). However, older adults did not judge the preventative relationship to be stronger when the response and outcome were separated by a short delay or when the outcome immediately followed their response. These findings point to a fundamental age related decline in the acquisition of preventative causal contingencies that may be due to changes in the utilization of cues for the retrieval of absent events. PMID- 19299257 TI - Clinically integrated health care in the English NHS. PMID- 19299256 TI - Health and functioning among baby boomers approaching 60. AB - OBJECTIVE: To investigate whether the health and functioning of the Baby Boom generation are better or worse than those of previous cohorts in middle age. METHODS: Trend analysis of vital statistics and self-reports from the National Health Interview Survey for the 40-59 population. Specific outcomes (years of data): mortality (1982-2004); poor or fair health (1982-2006); nine conditions (1997-2006); physical functional limitations (1997-2006); and needing help with personal care, routine needs, or either (1997-2006). RESULTS: In 2005, the mortality rate of 59-year-olds, the leading edge of the Baby Boom, was 31% lower than that of 59-year-olds in 1982 (8.3 vs. 12.1 per 1,000). There was a similar proportional decline in poor/fair health, but the decline reversed in the last decade. From 1997 to 2006, the prevalence of reports of four conditions increased significantly, but this trend may reflect improvements in diagnosis and treatment. Functional limitations and need for help with routine needs were stable, but the need for help with personal care, while quite low, increased. DISCUSSION: Trends varied by indicator, period, and age. It is surprising that, given the socioeconomic, medical, and public health advantages of Baby Boomers throughout their lives, they are not doing considerably better on all counts. PMID- 19299258 TI - Unhealthy markets: financial crisis, fiscal crisis ... health care crisis? PMID- 19299259 TI - Tight budgetary control: a study of clinical department managers' perceptions in Swedish hospitals. AB - OBJECTIVE: The composition of clinical department managers in Swedish hospitals is changing; more non-doctors and women are entering managerial positions. In parallel, most hospitals face increased pressure to contain costs. This article presents a study of managers' perceptions of tightness of budgetary control and how their views vary systematically with personal characteristics and organizational conditions. METHOD: Data were collected through a postal survey in 2005 to 173 clinical department managers (response rate of 70%). Statistical analysis was performed by factor analysis and logistic regression. RESULTS: The data suggest that clinical department managers' perceptions of tight budgetary control were related to how long they had been in their current position, their profession (whether they were doctors or non-doctors) and their sex. Further, their perceptions could be explained by how close the managers' departments were to their budget targets. CONCLUSIONS: Perception of tight budgetary control by managers depends on both their personal characteristics and the financial situation of their departments. Differences between men and women, and doctors and non-doctors call for additional research about the possible impact of changes in the composition of clinical department managers on how budgetary responsibility is exercised. PMID- 19299260 TI - Feasibility and cost of obtaining informed consent for essential review of medical records in large-scale health services research. AB - OBJECTIVE: To evaluate the effectiveness and cost of obtaining consent for review of medical records within the passively observed non-intervention arm of a cluster randomized controlled trial, 'Comparison Arm for ProtecT'. METHODS: Two hundred and thirty men, who had been notified to the trial by cancer registries as having prostate cancer, were sent a consent form from their general practitioner or secondary care clinician. The consent rate of participants to the review of their medical records and the estimated costs of the process were evaluated. RESULTS: One hundred and seventy-nine men (84%: 95% CI = 78%, 89%) consented to have their medical notes reviewed at an estimated cost of pound123 (euro172, $248) per person. CONCLUSIONS: A high consent rate for review of medical notes is achievable but at a cost. There needs to be renewed debate about the automatic need for consent to review medical records where the chance of personal harm is negligible and the purpose of the review is to provide robust evidence to save lives, prevent needless suffering, and improve the effectiveness and efficiency of health care delivery. PMID- 19299261 TI - Experience of continuity of care of patients with multiple long-term conditions in England. AB - OBJECTIVES: To examine patients' experiences of continuity of care in the context of different long-term conditions and models of care, and to explore implications for the future organization care of long-term conditions. METHODS: Qualitative semi-structured interviews were carried out with 33 patients recruited from seven general practices in South London. Patients were selected who had one or more of the following long-term conditions: arthritis, coronary heart disease, stroke, hypercholesterolaemia, hypertension, diabetes mellitus or chronic obstructive pulmonary disease. RESULTS: Multiple morbidity was frequent and experiences of continuity were framed within patients' wider experiences of health care rather than the context of a particular diagnosis. Positive experiences of relational continuity were strongly associated with long-term GP-led or specialist-led care. Management continuity was experienced in the context of shared care in terms of transitions between professionals or organizations. Access and flexibility issues were identified as important barriers or facilitators of continuity. CONCLUSIONS: Across a range of long-term conditions, patients' experiences of health care can be understood in terms of nuanced understandings of relational and management continuity. Continuity experiences, meanings and expectations, as well as barriers and facilitators, are influenced by the model of care rather than type of condition. PMID- 19299262 TI - Impacts of case management for frail elderly people: a qualitative study. AB - OBJECTIVE: To assess the impacts of different forms of case management for people aged over 65 years at risk of unplanned hospital admission, in particular the impacts upon patients, carers and health service organization in English primary care; and, in these respects, compare the Evercare model with alternatives. METHODS: Multiple qualitative case studies comparing case management in nine English Primary Care Trusts which piloted the Evercare model of case management and four sites which implemented alternative forms of case management between 2003 and 2005. Data were obtained from 231 interviews with patients, carers and other key informants, and from content analysis of documents and observation of meetings. RESULTS: All the projects established functioning case management services, but none led to major service reorganization or savings elsewhere in the health care system. Many informants reported examples of admissions which case management had prevented, but overall hospital admissions did not significantly change, possibly due to increased case-finding. Patients and carers valued case management for improving access to health care, increasing psychosocial support and improving communication with health professionals. CONCLUSION: Case management was highly valued by patients and their carers, but there were few major differences in outcomes between Evercare and other models. PMID- 19299263 TI - Does the culture of a medical practice affect the clinical management of diabetes by primary care providers? AB - OBJECTIVES: The financing and organization of primary care in the United States has changed dramatically in recent decades. Primary care physicians have shifted from solo practice to larger group practices. The culture of a medical practice is thought to have an important influence on physician behavior. This study examines the effects of practice culture and organizational structure (while controlling for patient and physician characteristics) on the quality of physician decision-making. METHODS: Data were obtained from a balanced factorial experiment which employed a clinically authentic video-taped scenario of diabetes with emerging peripheral neuropathy. RESULTS: Our findings show that several key practice culture variables significantly influence clinical decision-making with respect to diabetes. Practice culture may contribute more to whether essential examinations are performed than patient or physician variables or the structural characteristics of clinical organizations. CONCLUSIONS: Attention is beginning to focus on physician behavior in the context of different organizational environments. This study provides additional support for the suggestion that organization-level interventions (especially focused on practice culture) may offer an opportunity to reduce health care disparities and improve the quality of care. PMID- 19299264 TI - Partnerships for knowledge exchange in health services research, policy and practice. AB - Within the health services research community there is a growing strength of feeling that ongoing partnerships between researchers and decision-makers are critically important to effective transfer and exchange of knowledge generated from health services research. A body of literature is emerging around this idea that favours a particular model of partnership based on decision-maker involvement in research. This model is also gaining favour among health research funding bodies internationally. We argue that it is premature for the health services community to privilege any particular model of partnership between researchers and decision-makers. Rather a diversity of models should be conceptualized, explored in theory and practice, and evaluated. We identify seven dimensions that could be used to describe and differentiate models of partnerships for knowledge exchange and illustrate how these dimensions could be applied to analysing partnerships, using three case studies from recent and ongoing health services research partnerships in Australia. PMID- 19299265 TI - What is a priority? AB - What does it mean to say that something is a 'priority'? Priority setting is used to balance competing claims for resources, but the nature of the exercise is ambiguous. The priorities which are claimed might be for time, resources, process, rights or service. The setting of priorities might refer to importance, relative value, precedence, special status or lexical ordering. And there are different ways of ranking priorities including simple ordering, optimization, triage and satisficing. There is a fundamental distinction between preference rankings and precedence rankings, which can lead to strongly different conclusions from the same initial information. Because there is no definitive understanding of what a priority is, there can be no authoritative formula for deciding between competing claims. PMID- 19299266 TI - Commentary on 'What is a priority?' by Paul Spicker. PMID- 19299268 TI - Health services research: the gradual encroachment of ideas. AB - There is increasing pressure on researchers and research funding bodies to demonstrate the value of research. Simple approaches, consistent with the biomedical paradigm, based on relating the cost of research to its supposed impact are being investigated and adopted in laboratory and clinical research. While this may be appropriate in such research areas, it should not be applied to health services research which aims to alter the ways policy-makers and managers think about health, disease and health care or, as John Maynard Keynes put it, 'the gradual encroachment of ideas'. By considering six fundamental assumptions about health care that have been successfully challenged and overturned over the past few decades, the profound and sustained impact of health services research can be demonstrated. The application of economic models of 'payback' would fail to recognize such contributions which, in turn, could threaten future funding of health services research. PMID- 19299269 TI - Myth: in health care, more is always better. PMID- 19299270 TI - Board Certification: a historic milestone in phlebology in the USA. PMID- 19299271 TI - Phlebolymphoedema/chronic venous lymphatic insufficiency: an introduction to strategies for detection, differentiation and treatment. PMID- 19299272 TI - A comparison of low-molecular-weight heparin and combined therapy of low molecular-weight heparin with an anti-inflammatory agent in the treatment of superficial vein thrombosis. AB - OBJECTIVES: The optimal treatment of superficial vein thrombosis (SVT) of the leg has not been determined yet. The aim of this study is to evaluate the efficacy of low-molecular-weight heparin (LMWH) as compared with combined therapy of LMWH with an anti-inflammatory agent in treatment of SVT. METHODS: Fifty patients with SVT of the greater saphenous vein were randomly assigned to be treated with Ca nadroparin 190 IUAxa/kg in a single dose or with same dose of Ca-nadroparin and 60 mg oral acemetacine twice daily for 10 days. The efficacy of the two treatments to relieve symptoms and signs was evaluated by using visual analogue scale. RESULTS: Significant improvements were achieved for both groups after the treatment in terms of all four symptoms (P < 0.001). Treatment scores were in favour of LMWH with anti-inflammatory agent treatment group to relieve all four symptoms. The most significant and remarkable results obtained were for the reduction of pain and local tenderness (P < 0.05). No patient experienced major complications or mortality in either group. CONCLUSION: The results of this study suggest that the combined therapy of LMWH with an anti-inflammatory agent is more effective than LMWH. It may be an important option in the standard treatment of SVT. PMID- 19299273 TI - Local versus general anaesthesia for varicose veins surgery: a prospective non randomized controlled trial. AB - OBJECTIVE: To compare the acceptability and outcome of primary varicose vein (VV) surgery (saphenofemoral or saphenopopliteal ligation/excision) under local anaesthesia (LA) with that of general anaesthesia (GA). METHODS: A non-randomized controlled trial of consecutive patients treated between April 2004 and March 2006 was performed. After complete informed consent individual patients were asked to select their preferred form of anaesthesia (LA or GA). Preoperative disease status and co-morbidities were recorded. Outcomes were assessed perioperatively and at six weeks and six months postoperatively using patient scoring systems including the Aberdeen varicose veins severity score (AVVSS). RESULTS: Seventy-two (LA 46[62%] and GA 26[38%]) patients participated; median (range) age was 48 (21-74) years versus 36 (21-59) years (P = 0.0164), respectively. All procedures were performed as day cases. Median postoperative pain scores for LA and GA did not differ at 12 hours (4 versus 4; P = 0.48) and four days (5 versus 6; P = 0.44). Median improvement in the AVVSS at six weeks and six months for LA and GA cohorts were 5.7 versus 6.1 (P = 0.875) and 6.5 versus 8.3 (P = 0.131), respectively. Overall patient satisfaction did not show any intergroup difference at six weeks. CONCLUSIONS: Surgical treatment of VV under LA can be performed safely with comparable results to GA in self-selected patients. PMID- 19299274 TI - The relationship between the saphenopopliteal junction and the common peroneal nerve: a cada-veric study. AB - OBJECTIVES: The variable anatomy of the short saphenous vein (SSV) and the potential failure to identify the saphenopopliteal junction (SPJ) contribute to an increased risk of damage to the common peroneal nerve (CPN) during surgical exploration. The aim of the present study was to determine the variation of the SPJ, its relationship to the CPN, and the relationship of both SPJ and CPN to defined anatomical landmarks. METHODS: Measurements of the distance between the SPJ and CPN, and the defined anatomical landmarks (fibula head, lateral joint space, lateral femoral epicondyle), were undertaken on 30 cadaveric limbs following careful dissection of the popliteal fossa. RESULTS: The level of SPJ termination was classified as low (below), normal (within 100 mm above) and high (more than 100 mm above), the lateral femoral epicondyle. Of the 30 limbs dissected, 70% of SPJs were normal, 23% low and 7% high. Direct measurement from the SPJ to anatomical landmarks showed a higher interquartile range (IQR) in low compared with normal terminations; however, the vertical distance from the SPJ to the fibula head showed an increase in IQR from low to normal terminations (7.1 14.2). The mean distances between the SPJ and CPN in low and normal terminations were 23.3 and 16.7 mm, respectively. Comparison of the IQR showed values very similar to low terminations having a slightly higher IQR compared with normal terminations (7.15-6.0). CONCLUSION: Significant anatomic variation was observed in the termination of the SSV, with 67% located within 66 mm above the lateral femoral epicondyle. The risk of damaging the CPN during saphenopopliteal ligation may be higher for SPJs located above the lateral femoral epicondyle because of the proximity of the two structures and variability of SPJ. PMID- 19299275 TI - Five-year results of incompetent perforator vein closure using TRans-Luminal Occlusion of Perforator. AB - In 2000, we developed a percutaneous method of treating incompetent perforator veins (IPV) using ultrasound-guided radiofrequency ablation (RFA), which we termed TRansluminal Occlusion of Perforator (TRLOP). OBJECTIVE: To audit the five year outcome of the TRLOP technique as indicated by the rate of IPV closure on duplex ultrasound (DUS). METHODS: Patients underwent DUS five years post-TRLOP. Experienced vascular technologists documented the presence of IPVs using a two co ordinate system, blinded to previous results. Results were then compared with preoperative scans. IPVs were classified as: closed; not closed/reopened; or de novo. Closed IPVs were defined as the absence of any IPV at or within 5 cm of a previous IPV in the vertical and horizontal plane. Any IPVs found outside the delineated area were defined as de novo IPVs. RESULTS: Of 67 patients invited, 37 attended DUS (55% participation rate; men to women ratio of 14:23, age 40-84; mean 64). Preoperative clinical, aetiological, anatomical and pathological classification: C2, 36.2%; C3, 27.6%; C4, 34.5%; C6, 1.7%. From 125 IPVs analysed, 101 were closed (81%), 24 were not closed/reopened (19%) and 14 de novo IPVs were found. DISCUSSION: Despite these results representing our learning curve for the procedure, we found TRLOP to be an effective treatment for IPVs. The closure rates described are comparable with the published clinical series data for subfascial endoscopic perforator surgery. PMID- 19299276 TI - Surgical treatment of severe chronic venous insufficiency caused by pulsatile varicose veins in a patient with tricuspid regurgitation. AB - A case of severe chronic venous insufficiency caused by pulsatile varicose veins in a 46-year-old man with tricuspid regurgitation is presented. Active venous leg ulcer complicated with recurrent venous bleeding and inefficacy of conservative management serve as indications for surgical treatment. This case demonstrates the possibility of radical surgical correction of pathological venous reflux by means of saphenofemoral ligation, foam sclerotherapy and subfascial endoscopic perforator surgery. PMID- 19299277 TI - Spontaneous thoracic duct cyst presenting as a left supraclavicular mass - report of a case and review of literature. AB - Spontaneous and asymptomatic supraclavicular thoracic duct cysts occurring in the neck are the rarest. We report a case of a huge thoracic duct cyst occurring in the supraclavicular fossa with a review of the literature and management options. A 28-year-old female had a left supraclavicular mass with a 10-year history. A cervical thoracic duct cyst was diagnosed after chemical analysis of the milky fluid obtained by preoperative fine-needle aspiration. Treatment was achieved by surgical excision of the cyst and thoracic duct ligation. Pathological analysis of the cyst after excision confirmed the diagnosis. In the case of left supraclavicular masses, the rare differential diagnosis of thoracic duct cysts must be considered as a possibility. Magnetic resonance imaging as the imaging method of choice and chemical analysis of the milky fluid obtained by preoperative fine-needle aspiration are sufficient for primary diagnosis. The treatment of this disease consists of simple excision. PMID- 19299278 TI - Regarding endovenous laser ablation: mechanism of action. PMID- 19299279 TI - Men family caregivers' experience of nonsupportive interactions: context and expectations. AB - Men's involvement as family caregivers has grown as the prevalence of dementia has increased. Men rely on support from others for caregiving but also experience nonsupportive interactions. The purpose of this ethnographic study of 34 men (24 spouses and 10 sons) caring for a relative with dementia, 5 assisting caregivers, and 15 professionals was to identify primary caregivers' perceptions of nonsupportive and supportive interactions in relationships with kin and friends as well as professionals. Thematic analysis of transcribed data generated from interviews, diaries, and focus group discussions revealed the nature of men's caregiving journeys, the characteristics of their social networks, and their expectations of supportive interactions. The nonsupportive interactions men caregivers experienced included a lack of orientation to the caregiving situation, an unsatisfactory linkage to support sources, insufficient support, and hurtful interactions. Information about nonsupportive interactions can sensitize kin and friends as well as professionals to the complexity of men's experience and potentially avoid unintended negative consequences of support efforts. PMID- 19299280 TI - Help with overseas surgical training. PMID- 19299281 TI - Managing mycetomas. AB - Mycetomas are chronic, granulomatous, subcutaneous infections caused by traumatic inoculation into the skin of either the actinomycetes bacteria or the eumycetes fungi, giving rise to actinomycetomas and eumycetomas, respectively. They are endemic in the tropics afflicting mainly those of low socio-economic status and men working in agriculture. The disease is slowly progressive and can cause bone involvement, which can result in considerable disability. Late presentation is not uncommon making them notoriously difficult to manage. This article highlights the important aspects of their management and developments in drug therapy. PMID- 19299282 TI - Office haemorrhoidectomy with local anaesthesia: a feasible day-case procedure. AB - We report on 100 patients (88 men and 12 women) who underwent ligation-excision haemorrhoidectomy for prolapsing haemorrhoids in an outpatient office setting under local anaesthesia. Operating time, complications during procedure, postoperative complications and patient satisfaction with the procedure were analysed. The mean operating time was 55 (40-90) minutes. Ninety-seven were discharged three to five hours after surgery. Three patients were directly admitted to the hospital after surgery. One patient was readmitted after discharge. Severe postoperative pain occurred in eight; three suffered urinary retention; 88% were satisfied with the surgery but 12% were not. This study shows that, in selected patients, performing ligation-excision haemorrhoidectomy under local anaesthesia in the outpatient office setting is safe and viable. PMID- 19299283 TI - Decline in hospital mortality rate after the use of the World Health Organization protocol for management of severe malnutrition. AB - We studied the implementation of the World Health Organization protocol for the treatment of malnourished children at the largest maternal and infant hospital in the northeast of Brazil. The implementation of the protocol resulted in a reduction in the mortality rate from 38.0% to 16.2%. PMID- 19299284 TI - Do patients in rural Malawi benefit from upper gastrointestinal endoscopy? AB - The aim of this study was to evaluate the benefits of upper gastrointestinal endoscopy at central hospital level in Malawi and to draw conclusions from its use in the treatment of patients presenting with dysphagia and dyspepsia to health institutions in rural Africa. This retrospective study was carried out in order to investigate the endoscopic findings in 455 patients who presented to Zomba Central Hospital, Malawi, with upper gastrointestinal symptoms. Fifty-six percent of patients presenting with dysphagia were found to have oesophageal carcinoma. In 50% of patients with epigastric pain and 44% with abdominal pain, an endoscopy did not reveal any pathological findings. The intended treatment was frequently altered as a result of performing the endoscopy. We were able to treat patients according to their specific diagnosis in a significantly higher number than before: 51% versus 18% of those presenting with dysphagia, 96% versus 24% of those with epigastric pain and 90% versus 39% of those with abdominal pain. Upper gastrointestinal endoscopy is a feasible diagnostic tool in developing countries which improves the detection and treatment of diseases of the gastrointestinal tract. PMID- 19299285 TI - Atypical cerebrospinal fluid profile in tuberculous meningitis. AB - The aim of this study was to describe atypical cerebrospinal fluid (CSF) alterations in tuberculous meningitis (TBM) and to analyse the differences in outcome between patients with typical and atypical profiles. We did a retrospective study during the period of 2000 to 2005 including the cases of TBM assisted in a referral centre for infectious diseases in Rio de Janeiro State, Brazil. Neutrophilic plecytosis at the first spinal tap was found in 32.4% of TBM patients, who had a worse outcome when compared with those patients with typical CSF profiles. One factor that might have a major impact was the delay in starting empirical treatment (27.5 versus 11.6 days). We conclude that, in cases with clinical and epidemiological data compatible with TBM but with an atypical CSF profile, empirical treatment should be considered if CSF culture and direct examination for bacteria are negative. PMID- 19299286 TI - Peripheral neuropathy in HIV-positive patients at an antiretroviral clinic in Lilongwe, Malawi. AB - Peripheral neuropathy (PN) is common in the setting of antiretroviral (ARV) programmes in resource-limited settings and poses significant challenges in assessment and management. A retrospective analysis was undertaken of prevalence and management of PN in a cohort of 3341 patients on highly active antiretroviral therapy. A first line ARV regimen containing stavudine (D4T) is used for clinically eligible patients. Amitriptyline is prescribed for symptom relief and in cases of persistent or escalating symptoms zidovudine (AZT) is substituted for D4T. Leg pain or numbness was reported in 1173 patients (35%). However, only 428 (13%) were given a diagnosis of PN, 228 (7%) were prescribed amitriptyline and 200 (6%) were switched to AZT. A recent pharmokinetic study in this population showed a high Cmax of D4T with the generic combination triomune (D4T 40 mg). This could account for the high prevalence of PN. The optimum time for switch to a non D4T containing regimen is unknown. PMID- 19299287 TI - Presentation of trauma at the National Orthopaedic Hospital Enugu: a pilot study. AB - This prospective study spanning three months was undertaken to provide insight to trauma care including intervention times in this centre. Improved organization of trauma care is needed urgently for improved outcomes. PMID- 19299288 TI - Iron, folate and vitamin B12 parameters in HIV-1 infected patients with anaemia in southern Brazil. AB - We conducted a study to determine the role of iron, folate and vitamin B12 in HIV infected patients with anaemia attending a tertiary-care hospital in southern Brazil. Low serum folate levels were found in 14 (41%) HIV-infected patients; parameters of iron deficiency such as low transferring saturation index and ferritin in 10 (30%); and combined folate and iron deficiency in five (14%). Vitamin B12 deficiency was found in only two (6%) patients who presented with mean corpuscular volumes within the normal range. Our study has shown that folate and iron deficiency were frequently detected in HIV-infected patients at our institution, and should be considered in the differential diagnosis of anaemia in all HIV-infected patients independent of their HIV stage of progression. PMID- 19299289 TI - Clinical characteristics and outcome of Brucella endocarditis. AB - Brucella endocarditis, a rare complication of brucellosis, is the main cause of death attributable to this disease. There are difficulties in the diagnosis and uncertainty regarding many aspects of the treatment of Brucella endocarditis. We retrospectively examined the clinical characteristics and outcome of patients diagnosed with Brucella endocarditis. Of the six patients diagnosed as having Brucella endocarditis, four had valvular disease, one had aortic and mitral mechanic valve prosthesis (AVR+MVR) and one had secundum type atrial septal defect. Transesophageal echocardiography showed vegetations in four patients. Blood culture grew Brucella mellitensis only in two patients. Standard agglutination tests were elevated in all patients (range 1/320-1/10240). Four patients were managed with combined antibiotherapy and surgery. One refused further treatment and one refused an operation and follow-up was lost for that patient. Two patients died during follow-up; one having had a previous AVR+MVR operation refused further treatment and the other suffering renal failure. Due to the fulminant course of the disease, treatment should be initiated when there is a clinical suspicion, even if the culture results are unknown or negative. Agglutination titres aid in the diagnosis. A combination of antibiotherapy and surgery seems to be preferable treatment modality. PMID- 19299290 TI - False negative serological tests may lead to misdiagnosis and mismanagement in osteoarticular brucellosis. AB - Non-specific clinical presentations and paraclinical findings in osteoarticular brucellosis may mimic many other diseases and other causes of arthritis which could lead to misdiagnosis and mismanagement. We studied retrospectively the records of 232 patients with osteoarticular brucellosis who were admitted to the three teaching hospitals of Babol Medical Sciences University from April 2001 to September 2006. The distribution of osteoarticular involvement and their management were evaluated to determine if any cases had been misdiagnosed because of false negative serologic tests and who had, as a result, undergone inappropriate surgical interventions. Of 232 patients, 138 (59%) were male and 94 (41%) were female. Polyarthritis, monoarthritis, spondylitis and sacroilitis were seen in 91 (39%), 60 (26%), 43 (18.5%) and 38 (16.5%) patients, respectively. Two hundred and twenty-nine (98.7%) patients were diagnosed correctly and treated successfully; three (1.3%) were misdiagnosed and had been given inappropriate surgical interventions. Of these three patients, two (3.3%) were suffering from monoarthritis (hip joint) and one (2.3%) had spondylitis. PMID- 19299291 TI - Diagnosis of malaria in children's outpatient departments in Abuja, Nigeria. AB - Over-diagnosis of malaria has previously been described, especially in East Africa. Abuja is the capital of the most populous country in Africa. Over prescription of artemisinin-based combination therapy (ACT) antimalarials in Nigeria on the scale reported from other countries will result in substantial unnecessary use of this class of drug. In a study of 1000 children seen in the outpatient paediatric clinics of four district hospitals in Abuja, 669 had history of fever, of whom 616 (92%) were diagnosed with malaria. Only 24 (3.8%) where confirmed by positive malaria slides. Of 82 malaria tests requested, 32 (40%) were not available when clinicians wrote their prescriptions. Of 256 children prescribed an ACT, 11 (4.4%) were test negative, eight (3.1%) test positive, in 11 (4.3%) the test was not available and the test was not requested for 219 (86%). The proportion of available negative slides for patients treated with an antimalarial was 23/26 (88%), which was similar to the 22/24 (91%) treated with an antimalarial who were test positive. Testing for malaria made almost no impact on ACT prescription or on all other antimalarials and antibiotics. Based on these findings there is high possibility of massive over prescription of antimalarials. PMID- 19299292 TI - Surgical training in resource-limited countries: moving from the body to the bench--experiences from the basic surgical skills workshop in Enugu, Nigeria. AB - Increasingly, many institutions and surgical groups are resorting to laboratory simulations when teaching surgical skills. The Association of Surgeons of Great Britain and Ireland partnered with the West African College of Surgeons and Johnson and Johnson Corporation to introduce basic surgical skills (BSS) training into West Africa. The local faculty at the University of Nigeria Teaching Hospital, Enugu, was able to use the opportunity of this partnership to establish a regular basic surgical skills training programme. The achievement in Enugu shows that, with a dedicated team of local faculties, giving the local medical trainers a short introduction to the practicalities of organizing and conducting BSS is enough to jump start the programme in resource-poor countries. The Enugu example is a model that should be emulated by centres in other resource-poor countries to make the course a regular feature of their surgical training. PMID- 19299293 TI - The use of Foley's catheter in the removal of a coin in the oesophagus. AB - A coin in the oesophagus is a common paediatric emergency in Zambia. There are very few specialist surgeons and hospitals where endoscopic removal can be carried out in many resource-limited countries. The technique described here allows the removal of an ingested coin using an ordinary Foley's catheter. The use of this technique under fluoroscopic guidance has been described in literature. However, fluoroscopic facilities and trained radiologists are not commonly available in many parts of Africa. This technique is simple, safe and has a success rate close to that achieved by endoscopic removal. PMID- 19299294 TI - Kawasaki disease: a clinical and epidemiological study of 70 children in Brazil. AB - This is a prospective study of a cohort of 70 children with Kawasaki disease (KD) admitted from April 2002 to April 2007 to a tertiary hospital in Brasilia, Brazil. Of the 70 children, only 32 (45.7%) were referred during the acute phase of the disease and only 15 (21.4%) were referred with the correct diagnosis. Coronary aneurysms were detected in 13 (18.5%). A high suspicion index is essential in order to detect new cases as KD is probably underdiagnosed in Brazil and other developing countries, and an early diagnosis will dramatically decrease its life-threatening complications. PMID- 19299295 TI - Acalculous cholecystitis in a patient with Plasmodium falciparum infection after a trip to the Dominican Republic. AB - Acalculous cholecystitis has been associated with several infectious agents, but its relation with Plasmodium falciparum infection has not been clearly defined. This is the first case of acalculous cholecystitis produced by Plasmodium falciparum infection that is directly documented and should be included among the differential diagnoses of acalculous cholecystitis. PMID- 19299296 TI - Neurological disorders in rural Africa: a systematic approach. AB - Empirical knowledge suggests that neurological disorders are common in sub Saharan Africa. The aims of our study were to assess the hospital-based prevalence of neurological disorders in a rural African setting and to suggest a systematic approach to disease classification. Of 8676 admissions (over a period of eight months) 740 patients (8.5%) were given a neurological diagnosis; cases were grouped according to diagnostic certainty. We suggest three major categories for neurological disorders (group 1=no diagnostic uncertainties; group 2=minor diagnostic uncertainties; group 3=major diagnostic uncertainties) with clinical implications. PMID- 19299297 TI - Complications of paediatric elbow trauma treatment by traditional bonesetters. AB - This is a retrospective review of paediatric elbow trauma, which was initially treated by bonesetters and subsequently reported to the hospital for management. This paper describes the pattern of trauma and the complications of unscientific management. The report also recommends a basic training program for the bonesetters so as to make them aware of the potential complications involved in managing paediatric elbow trauma. PMID- 19299298 TI - Brucellosis in central Anatolia: evaluation of complications and relapse. AB - Brucellosis is an infectious disease involving many organs and tissues. We investigated retrospectively the brucellosis cases at our hospital, in order to study the relationship between clinical, laboratory and therapy findings, and relapse rates. We found that relapse was related to a positive family history, living in a rural area and the presence of complications. PMID- 19299299 TI - Atypical bacterial pathogens in community-acquired pneumonia in children: a hospital-based study. AB - A total of 243 children aged one month to five years with World Health Organization defined severe community acquired pneumonia were studied for the presence of atypical bacterial pathogens: 24 were found positive for mycoplasma infection. There was no significant association with any of the clinical, laboratory and radiological variables in children with pneumonia by the atypical pathogen. PMID- 19299300 TI - Tuberculous mycotic aneurysm of aortic root: an unusual cause of cardiac tamponade. AB - We describe a young male who presented to the emergency room with sudden onset dyspnea, and was found to have aortic root aneurysm with aortic regurgitation and cardiac tamponade. He underwent a Bentall procedure, and excised aortic root tissue showed epithelioid cell granulomas with panaortitis. He was started on anti-tubercular therapy, with which he improved. Although tubercular aortitis is fairly common, tuberculous mycotic aneurysm of the aorta is rare, with involvement of the aortic root being exceedingly uncommon. We report only the fifth case in English literature of tuberculous mycotic aneurysm of the aortic root. PMID- 19299301 TI - Ascariasis-associated worm encephalopathy in a young child. AB - Infestation with Ascaris lumbricoides in children has a varied manifestation, but encephalopathy is a very rare presentation. This report describes a case of ascariasis-associated encephalopathy in a child. An 18-month-old boy was admitted with altered sensorium. He had a history of vomiting and was passing Ascaris worms in the vomitus. The cerebrospinal fluid analysis did not reveal any abnormality. The patient was treated with an antihelminthic drug and he recovered completely. Worm encephalopathy should be considered as a differential diagnosis for unexplained encephalopathy in tropical areas. PMID- 19299302 TI - Disseminated cryptococcosis in an HIV-negative patient. AB - A young, unmarried, immunocompetent female presented with complaints of fever and altered sensorium, along with lesions over the face and trunk. A provisional diagnosis of disseminated herpes simplex with giant molluscum contagiosum was made. However, cutaneous biopsy of the lesions showed numerous cryptococcal organisms which stained positive to periodic acid-Schiff. This case is reported to sensitise physicians to keep cryptococcosis as a differential diagnosis in such types of skin lesions, as the lesions resemble that of molluscum contagiosum very closely. PMID- 19299303 TI - Dengue encephalitis: an entity now common in dengue-prone regions. AB - As well as dengue fever (DF) and dengue haemorrhagic fever-dengue shock syndrome (DHF/DSS), other atypical manifestations of dengue virus infection have also been reported. The frequency of CNS involvement in dengue remains unknown, although isolated cases with neurological manifestations have been reported in Southeast Asia, Malaysia, Burma, Puerto Rico and India. We present two cases of encephalitis associated with DF and DHF from New Delhi, India. PMID- 19299304 TI - Endomyocardial fibrosis: an enigmatic cause for a giant left atrium. AB - Endomyocardial fibrosis (EMF) was first described in 1948. It is an idiopathic disorder, characterized by the formation of fibrous tissue on the endocardial aspect of one or both ventricles, resulting in a restrictive cardiomyopathy with atrial enlargement. Originally described in Uganda, numerous cases have also been described in the Middle East. PMID- 19299305 TI - Total knee replacement--the evolving sub-Saharan experience. AB - Patients with moderate to severe osteoarthritis of the knee underwent primary elective knee arthroplasty in a well-equipped hospital in Accra, Ghana. Our main outcome measures was the new modified Oxford Knee scores (preoperative and postoperative scores). Median preoperative and postoperative knee scores were 10 and 42.5, respectively, at last follow-up with a P value<0.05 showing the reliability and significance of the scores. The average duration of inpatient hospital stay was 10 days. Of the seven patients who had surgery, one patient developed a chest infection postoperatively and another had failure of bone graft and required a revision. There was no postoperative wound infection or joint instability. The clinical outcome for this series of patients among other factors has been good within the prevailing circumstances and this shows that knee arthroplasty in developing Africa is a reality in the horizon. PMID- 19299306 TI - Can brucella endocarditis be treated successfully with medical therapy alone? AB - Human brucellosis is caused by one of the three species of Brucella: Brucella mellitensis, B. abortus, and B. suis. Worldwide, the incidence of human brucellosis ranges from <0.01 to >200 per 100,000 population. Cardiovascular complications occur in <2%, but accounts for most of the mortality. Brucella endocarditis usually involves normal native aortic valves in 75% of cases. A combination of antibiotics and valve replacement is the most acceptable treatment. So far only 15 adult cases have been reported as having been cured with medical therapy alone. We report another case treated successfully with antibiotics alone. PMID- 19299307 TI - Psoas abscess due to brucellosis. AB - Skeletal system involvement is a relatively common complication of human brucellosis. However, muscular involvement and psoas abscess are less frequently seen. Psoas abscess is usually secondary to spondylitis. We report on a 45-year old man and a 51-year-old woman with brucellosis complicated with spondylitis and psoas abscess. The man was successfully treated with the triple antibiotic regimen with percutaneous drainage of abscess. The woman was treated using an antibiotic combination. PMID- 19299308 TI - Plasmodium vivax malaria presenting as the nephrotic syndrome in an infant. PMID- 19299309 TI - Risk factors associated with major cerebrovascular complications after intracranial stenting. AB - BACKGROUND: There are limited data on the relationship between patient and site characteristics and clinical outcomes after intracranial stenting. METHODS: We performed a multivariable analysis that correlated patient and site characteristics with the occurrence of the primary endpoint (any stroke or death within 30 days of stenting or stroke in the territory of the stented artery beyond 30 days) in 160 patients enrolled in this stenting registry. All patients presented with an ischemic stroke, TIA, or other cerebral ischemic event (e.g., vertebrobasilar insufficiency) in the territory of a suspected 50-99% stenosis of a major intracranial artery while on antithrombotic therapy. RESULTS: Cerebral angiography confirmed that 99% (158/160) of patients had a 50-99% stenosis. In multivariable analysis, the primary endpoint was associated with posterior circulation stenosis (vs anterior circulation) (hazard ratio [HR] 3.4, 95% confidence interval [CI] 1.2-9.3, p = 0.018), stenting at low enrollment sites (< 10 patients each) (vs high enrollment site) (HR 2.8, 95% CI 1.1-7.6, p = 0.038), < or = 10 days from qualifying event to stenting (vs > or = 10 days) (HR 2.7, 95% CI 1.0-7.8, p = 0.058), and stroke as a qualifying event (vs TIA/other) (HR 3.2, 95% CI 0.9-11.2, p = 0.064). There was no significant difference in the primary endpoint based on age, gender, race, or percent stenosis (50-69% vs 70-99%). CONCLUSIONS: Major cerebrovascular complications after intracranial stenting may be associated with posterior circulation stenosis, low volume sites, stenting soon after a qualifying event, and stroke as the qualifying event. These factors will need to be monitored in future trials of intracranial stenting. PMID- 19299310 TI - Congenital muscular dystrophies with defective glycosylation of dystroglycan: a population study. AB - BACKGROUND: Congenital muscular dystrophies (CMD) with reduced glycosylation of alpha-dystroglycan (alpha-DG) are a heterogeneous group of conditions associated with mutations in six genes encoding proven or putative glycosyltransferases. OBJECTIVES: The aim of the study was to establish the prevalence of mutations in the six genes in the Italian population and the spectrum of clinical and brain MRI findings. METHODS: As part of a multicentric study involving all the tertiary neuromuscular centers in Italy, FKRP, POMT1, POMT2, POMGnT1, fukutin, and LARGE were screened in 81 patients with CMD and alpha-DG reduction on muscle biopsy (n = 76) or with a phenotype suggestive of alpha-dystroglycanopathy but in whom a muscle biopsy was not available for alpha-DG immunostaining (n = 5). RESULTS: Homozygous and compound heterozygous mutations were detected in a total of 43/81 patients (53%), and included seven novel variants. Mutations in POMT1 were the most prevalent in our cohort (21%), followed by POMT2 (11%), POMGnT1 (10%), and FKRP (9%). One patient carried two heterozygous mutations in fukutin and one case harbored a new homozygous variant in LARGE. No clear-cut genotype-phenotype correlation could be observed with each gene, resulting in a wide spectrum of clinical phenotypes. The more severe phenotypes, however, appeared to be consistently associated with mutations predicted to result in a severe disruption of the respective genes. CONCLUSIONS: Our data broaden the clinical spectrum associated with mutations in glycosyltransferases and provide data on their prevalence in the Italian population. PMID- 19299311 TI - Cost-effectiveness of preventive treatment of intracranial aneurysms: new data and uncertainties. AB - BACKGROUND: Previous modeling studies on treatment of unruptured intracranial aneurysms largely disregarded detailed data on treatment risks and omitted several factors that could influence cost-effectiveness. We performed a cost effectiveness analysis of surgical and endovascular treatment of unruptured aneurysms for different rupture rates and life expectancies, and assessed the influence of excess mortality risks in these persons, de novo development of aneurysms, and utility of awareness of having an untreated aneurysm, and also identified important factors for which data are lacking. METHODS: We used a Markov model to compare surgical, endovascular, and no treatment of unruptured intracranial aneurysms. Inputs for the model were taken mainly from meta analyses. Direct medical costs were derived from Dutch cost studies and expressed in 2005 Euros. We performed sensitivity analyses to evaluate model robustness. RESULTS: For 50-year-old patients, treatment of unruptured aneurysms is cost effective for all rupture rate scenarios between 0.3% and 3.5%/year. In 70-year old patients, treatment is not cost-effective in men with rupture rates < or =1%/year and women with rupture rates < or =0.5%/year. With lower utility of awareness of an untreated aneurysm, the cost-effectiveness of treatment strongly increased. The effect of excess mortality risks on the incremental cost effectiveness ratios was modest. The risk of formation of new aneurysms had no relevant impact. CONCLUSIONS: Patients' life expectancy, risk of rupture, and utility of awareness of an untreated aneurysm mainly define cost-effectiveness. However, important uncertainties remain on the rupture risk according to size and location of the aneurysm and on the utility of awareness of untreated aneurysm. More data on these factors are needed to define and individualize cost effectiveness analyses. PMID- 19299312 TI - Select nutrients in the ovine uterine lumen. VI. Expression of FK506-binding protein 12-rapamycin complex-associated protein 1 (FRAP1) and regulators and effectors of mTORC1 and mTORC2 complexes in ovine uteri and conceptuses. AB - FRAP1 (FK506-binding protein 12-rapamycin complex-associated protein 1), a component of the nutrient-sensing cell signaling pathway, is critical for cell growth and metabolism. The present study determined expression of FRAP1 and associated members of the mTORC1 and mTORC2 cell signaling pathways in uteri of cyclic and pregnant ewes and conceptuses, as well as effects of pregnancy, progesterone (P4), and interferon tau (IFNT) on their expression. The mRNAs for FRAP1, LST8, MAPKAP1, RAPTOR, RICTOR, TSC1, TSC2, RHEB, and EIF4EBP1 were localized to luminal, superficial glandular, and glandular epithelia and stromal cells of uteri from cyclic and pregnant ewes, as well as trophectoderm and endoderm of conceptuses between Days 13 and 18 of pregnancy. The abundance of FRAP1, RAPTOR, RICTOR, TSC1, and TSC2 mRNAs in endometria was unaffected by pregnancy status or by day of the estrous cycle or pregnancy; however, levels of LST8, MAPKAP1, RHEB, and EIF4EBP1 mRNA increased in endometria during early pregnancy. In ovariectomized ewes, P4 and IFNT stimulated expression of RHEB and EIF4EBP1 in uterine endometria. Total endometrial FRAP1 protein and phosphorylated FRAP1 protein levels were affected by pregnancy status and by day after onset of estrus, and phosphorylated FRAP1 protein was detected in nuclei of uterine epithelia and conceptuses. In endometria of pregnant ewes, increases in abundance of mRNAs for RICTOR, RHEB, and EIF4EBP1, as well as RHEB protein, correlated with rapid conceptus growth and development during the peri implantation period. These results suggest that the FRAP1 cell signaling pathway mediates interactions between the maternal uterus and peri-implantation conceptuses and that P4 and IFNT affect this pathway by regulating expression of RHEB and EIF4EBP1. PMID- 19299313 TI - Atrazine inhibits pulsatile luteinizing hormone release without altering pituitary sensitivity to a gonadotropin-releasing hormone receptor agonist in female Wistar rats. AB - Atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-tri-azine] is one of the most commonly used herbicides in the United States. Atrazine has been shown to suppress luteinizing hormone (LH) release and can lead to a prolongation of the estrous cycle in the rat. The objectives of this study were to examine the effects of atrazine on normal tonic release of LH and to elucidate the site of action of atrazine in the hypothalamic-pituitary-gonadal axis. Episodic release of gonadotropin-releasing hormone (GnRH) and the corresponding release of LH from the anterior pituitary gland are required for normal reproductive function. To determine if atrazine affects pulsatile LH release, ovariectomized adult female Wistar rats were administered atrazine (50, 100, or 200 mg/kg of body weight daily by gavage) or vehicle control for 4 days. On the final day of atrazine treatment, blood samples were obtained using an indwelling right atrial cannula. In the group receiving 200 mg/kg, there was a significant reduction in LH pulse frequency and a concomitant increase in pulse amplitude. To determine if the effects of atrazine on LH release were due to changes at the level of the pituitary, animals were passively immunized against endogenous GnRH, treated with atrazine, and challenged with a GnRH receptor agonist. Atrazine failed to alter pituitary sensitivity to the GnRH receptor agonist at any dose used. Taken together, these findings demonstrate that high doses of atrazine affect the GnRH pulse generator in the brain and not at the level of gonadotrophs in the pituitary. PMID- 19299314 TI - Regulation of sterol regulatory element-binding transcription factor 1a by human chorionic gonadotropin and insulin in cultured rat theca-interstitial cells. AB - Theca-interstitial (T-I) cells of the ovary synthesize androgens in response to luteinizing hormone (LH). In pathological conditions such as polycystic ovarian syndrome, T-I cells are hyperactive in androgen production in response to LH and insulin. Because cholesterol is an essential substrate for androgen production, we examined the effect of human chorionic gonadotropin (hCG) and insulin on signaling pathways that are known to increase cholesterol accumulation in steroidogenic cells. Specifically, the effect of hCG and insulin on sterol regulatory element-binding transcription factor 1a (SREBF1a) required for cholesterol biosynthesis and uptake was examined. Primary cultures of T-I cells isolated from 25-day-old rat ovaries responded to hCG and insulin to increase the active/processed form of SREBF1a. The hCG and insulin significantly reduced insulin-induced gene 1 (INSIG1) protein, a negative regulator of SREBF processing. Furthermore, an increase in the expression of selected SREBF target genes, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (Hmgcr) and mevalonate kinase (Mvk), was also observed. Protein kinase A (PRKA) inhibitor completely abolished the hCG-induced increase in SREBF1a, while increasing INSIG1. Although the hCG-induced depletion of total and free cholesterol was abolished by aminoglutethimide, the stimulatory effect on SREBF1a was not totally suppressed. Treatment with 25-hydroxycholesterol abrogated the effect of hCG on SREBF1a. Inhibition of the phosphatidylinositol 3-kinase pathway did not block the insulin induced increase in SREBF1a, whereas mitogen-activated protein kinase inhibition reduced the insulin response. These results suggest that the increased androgen biosynthesis by T-I cells in response to hCG and insulin is regulated, at least in part, by increasing the expression of sterol response element-responsive genes by increasing SREBF1a. PMID- 19299315 TI - Ciliary transport, gamete interaction, and effects of the early embryo in the oviduct: ex vivo analyses using a new digital videomicroscopic system in the cow. AB - Using a digital videomicroscopic analysis system in the bovine, we showed that the mechanisms of transport caused by ciliary beating are distinctly different in ampulla and isthmus of the oviduct. The average particle transport speed (PTS) in the oviduct (mean, 133 microm/sec) does not differ in the cycle (metestrus) and during pregnancy after implantation, but it is locally modulated at the site of the embryo. Using videomicroscopy, we were able to document that after entering the ampulla, the cumulus-oocyte complex (COC) is not transported by ciliary beating down the oviduct, but firmly attaches to the ampullar epithelium. This attachment is mediated by the cumulus cells. However, when a COC is degenerated, it is floating in the oviductal lumen. As soon as a vital COC is in the ampulla, the sperm bound in the sperm reservoir of the ampullar isthmic junction leave the reservoir and hurry to the oocyte. When a sperm has penetrated the zona pellucida, the COC detaches and continues its migration. Quantitative measurements showed that the early embryo is able to locally downregulate PTS during its migration down the oviduct. It locally changes the pattern of vascularization and induces the formation of secretory cells. Our studies imply that the oviductal epithelium is able to select vital oocytes. The early embryo is able to induce the formation of secretory cells, modify vascularization, and downregulate speed of transport, thus creating the prerequisite for the first embryo-maternal communication in the oviduct. PMID- 19299316 TI - Spermatogonial culture medium: an effective and efficient nutrient mixture for culturing rat spermatogonial stem cells. AB - An economical and simplified procedure to derive and propagate fully functional lines of undifferentiated rat spermatogonia in vitro is presented. The procedure is based on the formulation of a new spermatogonial culture medium termed SG medium. The SG medium is composed of a 1:1 mixture of Dulbecco modified Eagle medium:Ham F12 nutrient, 20 ng/ml of GDNF, 25 ng/ml of FGF2, 100 microM 2 mercaptoethanol, 6 mM l-glutamine, and a 1x concentration of B27 Supplement Minus Vitamin A solution. Using SG medium, six individual spermatogonial lines were derived from the testes of six separate Sprague-Dawley rats. After proliferating over a 120-day period in SG medium, stem cells within the spermatogonial cultures effectively regenerated spermatogenesis in testes of busulfan-treated recipient rats, which transmitted the donor cell haplotype to more than 75% of progeny by natural breeding. Subculturing in SG medium did not require protease treatment and was achieved by passaging the loosely bound spermatogonial cultures at 1:3 dilutions onto fresh monolayers of irradiated DR4 mouse fibroblasts every 12 days. Spermatogonial lines derived and propagated using SG medium were characterized as homogeneous populations of ZBTB16(+) DAZL(+) cells endowed with spermatogonial stem cell potential. PMID- 19299317 TI - Maturation, fertilization, and the structure and function of the endoplasmic reticulum in cryopreserved mouse oocytes. AB - Oocyte cryopreservation is a promising technology that could benefit women undergoing assisted reproduction. Most studies examining the effects of cryopreservation on fertilization and developmental competence have been done using metaphase II-stage oocytes, while fewer studies have focused on freezing oocytes at the germinal vesicle (GV) stage, followed by in vitro maturation. Herein, we examined the effects of vitrifying GV-stage mouse oocytes on cytoplasmic structure and on the ability to undergo cytoplasmic changes necessary for proper fertilization and early embryonic development. We examined the endoplasmic reticulum (ER) as one indicator of cytoplasmic structure, as well as the ability of oocytes to develop Ca(2+) release mechanisms following vitrification and in vitro maturation. Vitrified GV-stage oocytes matured in culture to metaphase II at a rate comparable to that of controls. These oocytes had the capacity to release Ca(2+) following injection of inositol 1,4,5 trisphosphate, demonstrating that Ca(2+) release mechanisms developed during meiotic maturation. The ER remained intact during the vitrification procedure as assessed using the lipophilic fluorescent dye DiI. However, the reorganization of the ER that occurs during in vivo maturation was impaired in oocytes that were vitrified before oocyte maturation. These results show that vitrification of GV stage oocytes does not affect nuclear maturation or the continuity of the ER, but normal cytoplasmic maturation as assessed by the reorganization of the ER is disrupted. Deficiencies in factors that are responsible for proper ER reorganization during oocyte maturation could contribute to the low developmental potential previously reported in vitrified in vitro-matured oocytes. PMID- 19299318 TI - Regulation of zebrafish zona pellucida gene activity in developing oocytes. AB - The two major zona pellucida proteins of the zebrafish chorion, Zp2 and Zp3, are encoded by multicopy genes arranged in tandem arrays on chromosomes 20 and 2, respectively. Expression of these zp genes in zebrafish is oocyte specific, and we report herein that their activity in developing oocytes is dependent on conserved CCAAT box sites in their promoters. A 140-bp region immediately upstream of the transcription initiation site (position 1) of the zp2 genes has been homogenized by gene conversion and contains a single CCAAT box located at 138 that is necessary for promoter activity in oocytes residing in stage I and early stage II ovarian follicles as determined by microinjection of promoter constructs linked to a luciferase reporter gene. The zp3 gene promoters have two inverted CCAAT boxes located in a region of shared homology within the initial 175 nucleotides. Serial deletion of these sites resulted in incremental decreases in luciferase activity. Double-stranded oligonucleotides containing CCAAT box sequences from both genes formed CCAAT box-specific complexes with ovarian follicle extracts in an electrophoretic mobility shift assay. We also found that the expression of the separate zebrafish zp3b gene, more closely related to two oocyte-expressed medaka zpc genes than to the tandemly arrayed zebrafish zp3 genes, is not CCAAT box dependent. The significance that these results have in furthering our understanding of the regulation of zebrafish zp gene evolution and regulation is discussed. PMID- 19299319 TI - Childhood victimization, poly-victimization, and adjustment to college in women. AB - This study examines the relationships among poly-victimization (i.e., high cumulative levels of victimization), six aggregate categories of childhood victimization (property crime, physical assault, peer and sibling, witnessed and indirect, sexual, child maltreatment), and college adjustment in females. This study first examines the relative contributions of poly-victimization and individual categories of childhood victimization in predicting college adjustment. The study then examines whether poly-victimization contributes any unique variance, beyond that accounted for by the combination of all six aggregate categories. Regression analyses reveal that a) poly-victimization accounts for a significant proportion of variability in scores for college adjustment, beyond that accounted for by any of the six categories of childhood victimization alone, and b) the categories of childhood victimization contribute little to no variability beyond that accounted for by poly-victimization. Furthermore, poly-victimization accounts for a significant proportion of variability in college adjustment, beyond that already accounted for by the simultaneous entry of all six categories as predictor variables. Finally, although victimization does not predict GPA, it predicts other domains of college adjustment. Results suggest that counselors working with college students should a) assess multiple categories of victimization and poly-victimization, and b) evaluate clients' adjustment to college across multiple domains (e.g., academic, social, interpersonal). PMID- 19299320 TI - Effect of "energy drink" consumption on hemodynamic and electrocardiographic parameters in healthy young adults. AB - BACKGROUND: Energy drinks are frequently purported to improve cognitive function and concentration. However, the cardiovascular effects of these drinks have not been adequately studied. OBJECTIVE: To determine the cardiac effects of a commercially available, multicomponent energy drink in healthy volunteers. METHODS: Fifteen healthy adults were included in this prospective study. Individuals who had chronic medical conditions, were on chronic medication, or were pregnant or breast-feeding were excluded. Subjects abstained from caffeine for 48 hours prior to and during the study. In the morning on Day 1 of the study, while subjects were in a fasted state, baseline blood pressure (BP), heart rate (HR), and electrocardiographic (ECG) parameters were measured. Participants then consumed 500 mL (2 cans) of an energy drink and measurements were repeated 30 minutes, 1 hour, 2 hours, 3 hours, and 4 hours later. Participants then drank 500 mL of energy drink daily for the next 5 days. Day 1 protocol was repeated on Day 7. RESULTS: On Days 1 and 7, maximum mean systolic BP (SBP), HR, and QTc interval occurred at 4 hours. Maximum diastolic BP (DBP) occurred at 2 hours on Days 1 and 7. Within 4 hours of energy drink consumption, on Days 1 and 7, respectively, SBP increased by 7.9% (p = 0.006) and 9.6% (p < 0.001), HR increased by 7.8% (p = 0.009) and 11.0% (p < 0.001), and QTc interval increased by 2.4% (p = 0.368) and 5.0% (p = 0.052). DBP increased by 7.0% (p = 0.046) and 7.8% (p = 0.063) within 2 hours of energy drink consumption on Days 1 and 7, respectively. CONCLUSIONS: Although no significant ECG changes were observed, HR increased 5-7 beats/min and SBP increased 10 mm Hg after energy drink consumption. PMID- 19299321 TI - Celecoxib-associated anaphylaxis. AB - OBJECTIVE: To report and discuss a case of anaphylaxis in a young, healthy white male taking celecoxib for intermittent lower back pain. CASE SUMMARY: A healthy 27-year-old man with a documented history of anaphylaxis to penicillins and macrolides presented to the emergency department (ED) in anaphylactic shock after ingesting a 200-mg capsule of celecoxib and a cup of orange juice. The patient had been taking celecoxib over the past 6 months, for 1-2 weeks at a time, for low back pain secondary to a pilonidal cyst and an L5/S1 bulging disc. The day of admission was the seventh day of the most recent course of twice-daily celecoxib. The patient initially self-treated the reaction with diphenhydramine and subcutaneous epinephrine that he had at home due to his history of drug- and bee sting-induced anaphylaxis; neither intervention improved his symptoms. He became profoundly diaphoretic and developed systemic swelling, shortness of breath, bradycardia, and hypotension. Emergency medical services transported the patient to the ED, where he was treated appropriately and the symptoms resolved. However, 4 hours later, at time of discharge from the ED, the symptoms recurred. He was admitted to the intensive care unit and monitored for 3 days. Supportive care, steroids, and histamine blockade provided resolution of the symptoms. Cardiac workup was initiated because of the recurrence and severity of bradycardia and hypotension; results of the workup were unremarkable. The patient was discharged in stable condition. DISCUSSION: This case demonstrates rare anaphylaxis to celecoxib in a patient who had previously taken the drug and who had documented tolerance to sulfonamide antibiotics. Despite this history, our patient developed type V immunoglobulin E-mediated anaphylaxis secondary to the sulfonamide component of celecoxib. This reaction was considered probable according to the Naranjo probability scale. A review of published case reports and related allergy literature for celecoxib allergenicity revealed that such reactions are rare. This is the first case report with great detail of a patient with anaphylaxis to celecoxib after having previously tolerated the medication. CONCLUSIONS: Celecoxib can produce an anaphylactic reaction in patients who have previously tolerated sulfonamide antibiotics and who have previously tolerated celecoxib. This case also reviews the potentially biphasic presentation of anaphylaxis. Clinicians need to be aware of this biphasic anaphylactic response to ensure optimal duration of evaluation. PMID- 19299322 TI - Lack of effect of Ginkgo biloba on voriconazole pharmacokinetics in Chinese volunteers identified as CYP2C19 poor and extensive metabolizers. AB - BACKGROUND: Ginkgo biloba is one of the most popular herbal supplements in the world. The supplement has been shown to induce the enzymatic activity of CYP2C19, the main cytochrome P450 isozyme involved in voriconazole metabolism. Because this enzyme exhibits genetic polymorphism, the inductive effect was expected to be modulated by the CYP2C19 metabolizer status. OBJECTIVE: To examine the possible effects of Ginkgo biloba as an inducer of CYP2C19 on single-dose pharmacokinetics of voriconazole in Chinese volunteers genotyped as either CYP2C19 extensive or poor metabolizers. METHODS: Fourteen healthy, nonsmoking volunteers-7 CYP2C19 extensive metabolizers (2C19(*)1/2C19(*)1) and 7 poor metabolizers (2C19(*)2/2C19(*)2)-were selected to participate in this study. Pharmacokinetics of oral voriconazole 200 mg after administration of Ginkgo biloba 120 mg twice daily for 12 days were determined for up to 24 hours by liquid chromatography-electrospray tandem mass spectrometry in a 2-phase randomized crossover study with 4-week washout between phases. RESULTS: For extensive metabolizers, the median value for voriconazole area under the plasma concentration-time curve from zero to infinity (AUC(0-)(infinity)) was 5.17 microg.h/mL after administration of voriconazole alone and 4.28 microg.h/mL after voriconazole with Ginkgo biloba (p > 0.05). The other pharmacokinetic parameters of voriconazole such as AUC(0-24), time to reach maximum concentration, half life, and apparent clearance also did not change significantly for extensive metabolizers in the presence of Ginkgo biloba. Pharmacokinetic parameters followed a similar pattern for poor metabolizers. CONCLUSIONS: The results suggest that 12 days of treatment with Ginkgo biloba did not significantly alter the single-dose pharmacokinetics of voriconazole in either CYP2C19 extensive or poor metabolizers. Therefore, the pharmacokinetic interactions between voriconazole and Ginkgo biloba may have limited clinical significance. PMID- 19299323 TI - Probable amlodipine-induced angioedema. AB - OBJECTIVE: To report a case of angioedema likely associated with amlodipine administration in a patient with a right thalamic hemorrhagic stroke. CASE SUMMARY: A 50-year-old female experienced angioedema during hospitalization for a right thalamic hemorrhagic stroke. She had no past history of angioedema and all of her medications were assessed for risk of angioedema. After careful evaluation, case reports linking calcium channel blockers (CCBs) and angioedema led to further examination of amlodipine as a cause. Amlodipine therapy had been initiated 24 hours prior to the development of angioedema, which then resolved 72 hours after discontinuation of the drug. In total, the patient experienced oropharyngeal swelling for 10 days. DISCUSSION: In determining a cause for the patient's angioedema we eliminated genetic, allergic, physically induced, thyroid autoimmune disease-associated, and medication-induced causes. Three case reports describing 7 patients have linked the CCBs verapamil, diltiazem, and nifedipine with angioedema. The onset and resolution of symptoms in our patient were very similar to those seen in other case reports. Application of the Naranjo probability scale found a probable link between amlodipine and angioedema. CONCLUSIONS: Although few reports of CCB-induced angioedema exist, to our knowledge, this is the first reported case to suggest a link between angioedema and amlodipine therapy. Clinicians should consider amlodipine as a potential cause of angioedema. PMID- 19299324 TI - Midodrine and octreotide in treatment of cirrhosis-related hemodynamic complications. AB - OBJECTIVE: To review studies evaluating the use of midodrine and octreotide in hemodynamic complications of cirrhosis, including ascites and hepatorenal syndrome. DATA SOURCES: Searches of MEDLINE (1966-September 2008) and EMBASE (1974-September 2008) were conducted using the terms midodrine, octreotide, hepatorenal syndrome, ascites, cirrhosis, and paracentesis-induced circulatory dysfunction. Literature review was limited to English-language, human studies. STUDY SELECTION AND DATA EXTRACTION: Studies identified from data sources were considered for review. Studies were excluded if primary therapy involved any of the following: transjugular intrahepatic portosystemic shunt procedure, medications other than midodrine or octreotide, or patients included for treatment or prevention of portal hypertension and/or variceal bleeding. Pharmacokinetic/pharmacodynamic studies and studies using retrospective data collection were excluded. Seven studies were included in this review. DATA SYNTHESIS: Midodrine and octreotide in combination or alone have shown conflicting results for systemic and renal hemodynamics and renal function in patients with cirrhosis-related complications. Patients with ascites being treated with midodrine, alone or in combination with octreotide, showed significant changes in systemic hemodynamics, without a correlating change in renal perfusion. Studies comparing the use of midodrine with use of albumin for the prevention of paracentesis-induced circulatory dysfunction (PICD) showed no incidence of PICD in either treatment group. In hepatorenal syndrome, patients using midodrine with octreotide showed significant changes in systemic hemodynamics and improvements in renal perfusion. This regimen's effect on survival is yet to be determined. CONCLUSIONS: Available evidence shows inconsistent results for the effectiveness and safety of midodrine and octreotide use in cirrhotic patients. Because of the contradictory results, longer treatment duration and increased number of study participants are necessary to determine the proper use of midodrine and octreotide in these patients. PMID- 19299325 TI - Neuroleptic malignant syndrome secondary to quetiapine. AB - OBJECTIVE: To report a case of neuroleptic malignant syndrome (NMS) secondary to quetiapine in which the patient developed extrapyramidal symptoms (EPS). CASE SUMMARY: A 34-year-old male with a history of severe brain damage, mental retardation, and seizures was admitted to the hospital with changes in mental status, development of tremors, and a temperature of 39.9 degrees C. Initial differential diagnoses included seizure, aspiration, stroke, and infection. Once these were excluded, NMS was considered. The patient exhibited other characteristics of NMS during hospitalization, including lead pipe rigidity, tachycardia, and high creatine kinase level (up to 12,654 IU/L). Drug therapy on presentation included quetiapine 200 mg 3 times per day, guanfacine 2 mg/day, carbamazepine 400 mg every 12 hours, valproic acid 500 mg twice daily, and lorazepam 2 mg (unknown schedule). He reportedly had received these medications for at least a month before admission. On hospital day 2, quetiapine was discontinued. The patient received traditional treatment for NMS, which included bromocriptine, dantrolene, intravenous fluids, and supportive care. The NMS resolved in 7 days. DISCUSSION: In cases of NMS, clinicians previously believed that the risk for developing severe adverse effects such as EPS was lower with atypical versus typical antipsychotics. We identified 13 cases of NMS secondary to quetiapine in the literature via a search of MEDLINE/PubMed (1950-2008), and Iowa Drug Information Service (1966-2008). Seventy-five percent of previous reports of NMS secondary to quetiapine had reactions that included EPS. Common patient characteristics in our report and others included male sex, history of mental retardation, and treatment modalities used in NMS. Unique characteristics in this case included length of therapy without dosage change or titration and no known history of drug-related EPS. The Naranjo probability scale indicated a probable relationship between the development of NMS and quetiapine. CONCLUSIONS: NMS with associated EPS has been previously associated with quetiapine. Clinicians should be aware that NMS with EPS can occur with quetiapine at steady state doses without recent dosage adjustments or titration. PMID- 19299326 TI - Effects of quetiapine on sleep in nonpsychiatric and psychiatric conditions. AB - OBJECTIVE: To evaluate the use of immediate-release quetiapine for the treatment of insomnia. DATA SOURCES: Pre-MEDLINE and MEDLINE were searched (1966 to October 2008) using the terms quetiapine, sleep, insomnia, and antipsychotics. STUDY SELECTION AND DATA EXTRACTION: All studies and case reports evaluating insomnia as a primary endpoint were reviewed. DATA SYNTHESIS: The role of quetiapine for improving sleep in various patient populations is uncertain. Quetiapine has moderately sedative properties, and doses used in treatment of insomnia have ranged from 12.5 to 800 mg. Results of clinical trials and observations in case studies have revealed possible beneficial effects of quetiapine on several subjective and objective sleep parameters. In most studies, significant improvements in sleep were found in areas of total sleep time, sleep efficiency, and subjective sleep scores. However, some of these results may not be clinically significant. Also, quetiapine has been found to have adverse effects such as periodic leg movements, akathisia, and metabolic complications. Additionally, changes in rapid eye movement (REM) and percentage of REM sleep have been noted in different populations and need further study. Despite quetiapine's sedative properties, current data do not appear to support its use as first-line treatment for sleep complications. However, it may be useful for treatment of insomnia in patients with psychiatric disorders (eg, bipolar, schizophrenia) who do not respond to primary or secondary treatments. CONCLUSIONS: Further studies are needed to define the placement, dose, and adverse effects of quetiapine for the treatment of sleep problems. PMID- 19299327 TI - The LDL receptor. AB - In this article, the history of the LDL receptor is recounted by its codiscoverers. Their early work on the LDL receptor explained a genetic cause of heart attacks and led to new ways of thinking about cholesterol metabolism. The LDL receptor discovery also introduced three general concepts to cell biology: receptor-mediated endocytosis, receptor recycling, and feedback regulation of receptors. The latter concept provides the mechanism by which statins selectively lower plasma LDL, reducing heart attacks and prolonging life. PMID- 19299328 TI - VEGF and restenosis: the rest of the story. PMID- 19299329 TI - Caspase-8, a double-edged sword for EPC functioning. PMID- 19299330 TI - Getting radical about obesity: new links between fat and heart disease. PMID- 19299331 TI - Generation of induced pluripotent stem cells from human blood. AB - Human dermal fibroblasts obtained by skin biopsy can be reprogrammed directly to pluripotency by the ectopic expression of defined transcription factors. Here, we describe the derivation of induced pluripotent stem cells from CD34+ mobilized human peripheral blood cells using retroviral transduction of OCT4/SOX2/KLF4/MYC. Blood-derived human induced pluripotent stem cells are indistinguishable from human embryonic stem cells with respect to morphology, expression of surface antigens, and pluripotency-associated transcription factors, DNA methylation status at pluripotent cell-specific genes, and the capacity to differentiate in vitro and in teratomas. The ability to reprogram cells from human blood will allow the generation of patient-specific stem cells for diseases in which the disease-causing somatic mutations are restricted to cells of the hematopoietic lineage. PMID- 19299332 TI - Selective expression of latency-associated peptide (LAP) and IL-1 receptor type I/II (CD121a/CD121b) on activated human FOXP3+ regulatory T cells allows for their purification from expansion cultures. AB - Although adoptive transfer of regulatory T cells (Foxp3(+) Tregs) has proven to be efficacious in the prevention and treatment of autoimmune diseases and graft versus-host disease in rodents, a major obstacle for the use of Treg immunotherapy in humans is the difficulty of obtaining a highly purified preparation after ex vivo expansion. We have identified latency-associated peptide (LAP) and IL-1 receptor type I and II (CD121a/CD121b) as unique cell surface markers that distinguish activated Tregs from activated FOXP3(-) and FOXP3(+) non-Tregs. We show that it is feasible to sort expanded FOXP3(+) Tregs from non-Tregs with the use of techniques for magnetic bead cell separation based on expression of these 3 markers. After separation, the final product contains greater than 90% fully functional FOXP3(+) Tregs. This novel protocol should facilitate the purification of Tregs for both cell-based therapies as well as detailed studies of human Treg function in health and disease. PMID- 19299333 TI - How we treat cytomegalovirus in hematopoietic cell transplant recipients. AB - Cytomegalovirus (CMV) continues to cause major complications after hematopoietic cell transplantation (HCT). Over the past decade, most centers have adopted preemptive antiviral treatment or prophylaxis strategies to prevent CMV disease. Both strategies are effective but also have shortcomings with presently available drugs. Here, we review aspects of CMV treatment and prevention in HCT recipients, including currently used drugs and diagnostics, ways to optimize preemptive therapy strategies with quantitative polymerase chain reaction assays, the use of prophylaxis, management of CMV disease caused by wild-type or drug-resistant strains, and future strategies. PMID- 19299334 TI - Reciprocal responsiveness to interleukin-12 and interferon-alpha specifies human CD8+ effector versus central memory T-cell fates. AB - Multiple innate signals regulate the genesis of effector and memory CD8+ T cells. In this study, we demonstrate that the innate cytokines interleukin (IL)-12 and interferon (IFN)-alpha/beta regulate distinct aspects of effector and memory human CD8+ T-cell differentiation. IL-12 exclusively promoted the development of IFN-gamma- and tumor necrosis factor (TNF)-alpha-secreting T effector memory (T(EM)) cells, whereas IFN-alpha drove the development of T central memory (T(CM)) cells. The development of T(EM) and T(CM) was linked to cell division. In rapidly dividing cells, IL-12 programmed T(EM) through induction of the IL-12 receptor beta2. In contrast, IFN-alpha regulated T(CM) development by slowing the progression of cell division in a subpopulation of cells that selectively expressed elevated IFN-alpha/beta receptor-2. The strength of signal delivered through T-cell receptor (TCR) engagement regulated the responsiveness of cells to IL-12 and IFN-alpha. In the presence of both IL-12 and IFN-alpha, these cytokine signals were amplified as the strength of the TCR signal was increased, promoting the simultaneous development of both T(CM) and T(EM). Together, our results support a novel model in which IL-12 and IFN-alpha act in a nonredundant manner to regulate the colinear generation of both effector and memory cells. PMID- 19299335 TI - Induction of angiogenesis by normal and malignant plasma cells. AB - Abundant bone marrow angiogenesis is present in almost all myeloma patients requiring therapy and correlated to treatment response and survival. We assessed the expression of 402 angiogenesis-associated genes by Affymetrix DNA microarrays in 466 samples, including CD138-purified myeloma cells (MMCs) from 300 previously untreated patients, in vivo microcirculation by dynamic contrast-enhanced magnetic resonance imaging, and in vitro angiogenesis (AngioKit-assay). Normal bone marrow plasma cells (BMPCs) express a median of 39 proangiogenic (eg, VEGFA, ADM, IGF-1) and 28 antiangiogenic genes (eg, TIMP1, TIMP2). Supernatants of BMPCs unlike those of memory B cells induce angiogenesis in vitro. MMCs do not show a significantly higher median number of expressed proangiogenic (45) or antiangiogenic (31) genes, but 97% of MMC samples aberrantly express at least one of the angiogenic factors HGF, IL-15, ANG, APRIL, CTGF, or TGFA. Supernatants of MMCs and human myeloma cell lines induce significantly higher in vitro angiogenesis compared with BMPCs. In conclusion, BMPCs express a surplus of proangiogenic over antiangiogenic genes transmitting to the ability to induce in vitro angiogenesis. Aberrant expression of proangiogenic and down-regulation of antiangiogenic genes by MMCs further increases the angiogenic stimulus, together leading to bone marrow angiogenesis at various degrees in all myeloma patients. PMID- 19299336 TI - Genome-wide association study to identify novel loci associated with therapy related myeloid leukemia susceptibility. AB - Therapy-related acute myeloid leukemia (t-AML) is a rare but fatal complication of cytotoxic therapy. Whereas sporadic cancer results from interactions between complex exposures and low-penetrance alleles, t-AML results from an acute exposure to a limited number of potent genotoxins. Consequently, we hypothesized that the effect sizes of variants associated with t-AML would be greater than in sporadic cancer, and, therefore, that these variants could be detected even in a modest-sized cohort. To test this, we undertook an association study in 80 cases and 150 controls using Affymetrix Mapping 10K arrays. Even at nominal significance thresholds, we found a significant excess of associations over chance; for example, although 6 associations were expected at P less than .001, we found 15 (P(enrich) = .002). To replicate our findings, we genotyped the 10 most significantly associated single nucleotide polymorphisms (SNPs) in an independent t-AML cohort (n = 70) and obtained evidence of association with t-AML for 3 SNPs in the subset of patients with loss of chromosomes 5 or 7 or both, acquired abnormalities associated with prior exposure to alkylator chemotherapy. Thus, we conclude that the effect of genetic factors contributing to cancer risk is potentiated and more readily discernable in t-AML compared with sporadic cancer. PMID- 19299337 TI - Integrin-linked kinase associated with integrin activation. AB - Platelet integrin alphaIIbbeta3 activation is tightly controlled by intracellular signaling pathways, and several molecules, including talin, have been identified as critical for alphaIIbbeta3 activation. However, the whole pathway associated with alphaIIbbeta3 activation remains to be determined. To address this issue, we established a Chinese hamster ovary cell line (parental cells) that expresses constitutively activated chimeric integrin alphaIIbalpha6Bbeta3, and then obtained mutant cells expressing inactivated alphaIIbalpha6Bbeta3 by genome-wide mutagenesis. We have performed expression cloning to isolate signaling molecules responsible for integrin activation in the mutant cells. We show that integrin linked kinase (ILK) complements defective integrin activation in the mutant cells. ILK mRNAs in the mutant cells contained 2 nonsense mutations, R317X and W383X, in a compound heterozygous state, resulting in a complete loss of ILK expression. Moreover, the mutant cells showed partially impaired activation of endogenous beta1 integrins. Knockdown of ILK in parental cells significantly suppressed the activated state of alphaIIbalpha6Bbeta3. However, ILK overexpression did not rescue the impaired integrin activation in talin knocked down parental cells, whereas overexpression of talin-F3, a subdomain of the talin head domain, restored the function. Our present data suggest that ILK contributes to inside-out integrin activation. PMID- 19299338 TI - Nucleotide polymorphism and within-gene recombination in Daphnia magna and D. pulex, two cyclical parthenogens. AB - Theory predicts that partially asexual organisms may make the "best of both worlds": for the most part, they avoid the costs of sexual reproduction, while still benefiting from an enhanced efficiency of selection compared to obligately asexual organisms. There is, however, little empirical data on partially asexual organisms to test this prediction. Here we examine patterns of nucleotide diversity at eight nuclear loci in continentwide samples of two species of cyclically parthenogenetic Daphnia to assess the effect of partial asexual reproduction on effective population size and amount of recombination. Both species have high nucleotide diversities and show abundant evidence for recombination, yielding large estimates of effective population sizes (300,000 600,000). This suggests that selection will act efficiently even on mutations with small selection coefficients. Divergence between the two species is less than one-tenth of previous estimates, which were derived using a mitochondrial molecular clock. As the two species investigated are among the most distantly related species of the genus, this suggests that the genus Daphnia may be considerably younger than previously thought. Daphnia has recently received increased attention because it is being developed as a model organism for ecological and evolutionary genomics. Our results confirm the attractiveness of Daphnia as a model organism, because the high nucleotide diversity and low linkage disequilibrium suggest that fine-scale mapping of genes affecting phenotypes through association studies should be feasible. PMID- 19299339 TI - Genomic selection using low-density marker panels. AB - Genomic selection (GS) using high-density single-nucleotide polymorphisms (SNPs) is promising to improve response to selection in populations that are under artificial selection. High-density SNP genotyping of all selection candidates each generation, however, may not be cost effective. Smaller panels with SNPs that show strong associations with phenotype can be used, but this may require separate SNPs for each trait and each population. As an alternative, we propose to use a panel of evenly spaced low-density SNPs across the genome to estimate genome-assisted breeding values of selection candidates in pedigreed populations. The principle of this approach is to utilize cosegregation information from low density SNPs to track effects of high-density SNP alleles within families. Simulations were used to analyze the loss of accuracy of estimated breeding values from using evenly spaced and selected SNP panels compared to using all high-density SNPs in a Bayesian analysis. Forward stepwise selection and a Bayesian approach were used to select SNPs. Loss of accuracy was nearly independent of the number of simulated quantitative trait loci (QTL) with evenly spaced SNPs, but increased with number of QTL for the selected SNP panels. Loss of accuracy with evenly spaced SNPs increased steadily over generations but was constant when the smaller number individuals that are selected for breeding each generation were also genotyped using the high-density SNP panel. With equal numbers of low-density SNPs, panels with SNPs selected on the basis of the Bayesian approach had the smallest loss in accuracy for a single trait, but a panel with evenly spaced SNPs at 10 cM was only slightly worse, whereas a panel with SNPs selected by forward stepwise selection was inferior. Panels with evenly spaced SNPs can, however, be used across traits and populations and their performance is independent of the number of QTL affecting the trait and of the methods used to estimate effects in the training data and are, therefore, preferred for broad applications in pedigreed populations under artificial selection. PMID- 19299340 TI - Conserved WCPL and CX4C domains mediate several mating adhesin interactions in Saccharomyces cerevisiae. AB - Several adhesins are induced by pheromones during mating in Saccharomyces cerevisiae, including Aga1p, Aga2p, Sag1p (Agalpha1p), and Fig2p. These four proteins all participate in or influence a well-studied agglutinin interaction mediated by Aga1p-Aga2p complexes and Sag1p; however, they also play redundant and essential roles in mating via an unknown mechanism. Aga1p and Fig2p both contain repeated, conserved WCPL and CX(4)C domains. This study was directed toward understanding the mechanism underlying the collective requirement of agglutinins and Fig2p for mating. Apart from the well-known agglutinin interaction between Aga2p and Sag1p, three more pairs of interactions in cells of opposite mating type were revealed by this study, including bilateral heterotypic interactions between Aga1p and Fig2p and a homotypic interaction between Fig2p and Fig2p. These four pairs of adhesin interactions are collectively required for maximum mating efficiency and normal zygote morphogenesis. GPI-less, epitope tagged forms of Aga1p and Fig2p can be co-immunoprecipitated from the culture medium of mating cells in a manner dependent on the WCPL and CX(4)C domains in the R1 repeat of Aga1p. Using site-directed mutagenesis, the conserved residues in Aga1p that interact with Fig2p were identified. Aga1p is involved in two distinct adhesive functions that are independent of each other, which raises the possibility for combinatorial interactions of this protein with its different adhesion receptors, Sag1 and Fig2p, a property of many higher eukaryotic adhesins. PMID- 19299341 TI - Linkage analysis reveals the independent origin of Poeciliid sex chromosomes and a case of atypical sex inheritance in the guppy (Poecilia reticulata). AB - Among different teleost fish species, diverse sex-determining mechanisms exist, including environmental and genetic sex determination, yet chromosomal sex determination with male heterogamety (XY) prevails. Different pairs of autosomes have evolved as sex chromosomes among species in the same genus without evidence for a master sex-determining locus being identical. Models for evolution of Y chromosomes predict that male-advantageous genes become linked to a sex determining locus and suppressed recombination ensures their co-inheritance. In the guppy, Poecilia reticulata, a set of genes responsible for adult male ornaments are linked to the sex-determining locus on the incipient Y chromosome. We have identified >60 sex-linked molecular markers to generate a detailed map for the sex linkage group of the guppy and compared it with the syntenic autosome 12 of medaka. We mapped the sex-determining locus to the distal end of the sex chromosome. We report a sex-biased distribution of recombination events in female and male meiosis on sex chromosomes. In one mapping cross, we observed sex ratio and male phenotype deviations and propose an atypical mode of genetic sex inheritance as its basis. PMID- 19299342 TI - Factors affecting accuracy from genomic selection in populations derived from multiple inbred lines: a Barley case study. AB - We compared the accuracies of four genomic-selection prediction methods as affected by marker density, level of linkage disequilibrium (LD), quantitative trait locus (QTL) number, sample size, and level of replication in populations generated from multiple inbred lines. Marker data on 42 two-row spring barley inbred lines were used to simulate high and low LD populations from multiple inbred line crosses: the first included many small full-sib families and the second was derived from five generations of random mating. True breeding values (TBV) were simulated on the basis of 20 or 80 additive QTL. Methods used to derive genomic estimated breeding values (GEBV) were random regression best linear unbiased prediction (RR-BLUP), Bayes-B, a Bayesian shrinkage regression method, and BLUP from a mixed model analysis using a relationship matrix calculated from marker data. Using the best methods, accuracies of GEBV were comparable to accuracies from phenotype for predicting TBV without requiring the time and expense of field evaluation. We identified a trade-off between a method's ability to capture marker-QTL LD vs. marker-based relatedness of individuals. The Bayesian shrinkage regression method primarily captured LD, the BLUP methods captured relationships, while Bayes-B captured both. Under most of the study scenarios, mixed-model analysis using a marker-derived relationship matrix (BLUP) was more accurate than methods that directly estimated marker effects, suggesting that relationship information was more valuable than LD information. When markers were in strong LD with large-effect QTL, or when predictions were made on individuals several generations removed from the training data set, however, the ranking of method performance was reversed and BLUP had the lowest accuracy. PMID- 19299343 TI - Diffraction-enhanced imaging for Achilles tendon lesions: a preliminary study. AB - BACKGROUND: Computed tomography, ultrasonography, and magnetic resonance imaging are useful in the diagnosis of tears of the Achilles tendon, but none are capable of detecting early or small tears. Herein, we applied diffraction-enhanced imaging, a radiographic technique that detects x-ray attenuation and x-ray refraction, to the imaging of compromised Achilles tendons. METHODS: Diffraction enhanced imaging was used to detect incomplete surgically induced tears of the Achilles tendon in nine cadaveric human feet and ankles. RESULTS: Complete and significant partial tears were detectable in diffraction-enhanced images as x-ray refraction changes. CONCLUSIONS: Although still in the experimental stages, diffraction-enhanced imaging may eventually prove useful for the diagnosis of Achilles tendon tears. PMID- 19299344 TI - A method for assessing off-loading compliance. AB - BACKGROUND: Off-loading excessive pressure is essential to healing diabetic foot ulcers. However, many patients are not compliant in using prescribed footwear or off-loading devices. We sought to validate a method of objectively measuring off loading compliance via activity monitors. METHODS: For 4 days, a single subject maintained a written compliance diary concerning use of a removable cast walker. He also wore a hip-mounted activity monitor during all waking hours. An additional activity monitor remained mounted on the cast walker at all times. At the conclusion of the 4 days, the time-stamped hip activity data were independently coded for walker compliance by the compliance diary and by using the time-stamped walker activity data. RESULTS: An intraclass reliability of 0.93 was found between diary-coded and walker monitor-coded activity. CONCLUSIONS: These results support the use of this dual activity monitor approach for assessing off-loading compliance. An advantage of this approach versus a patient maintained diary is that the monitors are not susceptible to incorrect patient recall or a patient's desire to please a caregiver by reporting inflated compliance. Furthermore, these results seem to lend support to existing reports in the literature using similar methods. PMID- 19299345 TI - Histologic evaluation of a 6-month GraftJacket matrix biopsy used for Achilles tendon augmentation. AB - BACKGROUND: Assessing implanted biological reinforcement graft success in soft tissue repairs is typically limited to noninvasive measurements and functional outcome measures. However, there are times when a histologic snapshot of the graft incorporation may be possible owing to a nongraft-related postoperative complication, such as hardware failure. METHODS: We histologically evaluated a 6 month biopsy sample from an Achilles tendon repair augmented with an acellular human dermal matrix (AHDM). A 57-year-old woman was treated for Haglund's deformity of the Achilles tendon. The Achilles tendon was fixed to the calcaneus using a plate, and an AHDM was used to augment the primary repair of the tendon. At 6 months, the hardware was removed owing to prominence, and a biopsy of the AHDM was performed. The specimen was prepared and stained using hematoxylin and eosin, Verhoeff-van Gieson, Movat's pentachrome, and toluidine blue stains. RESULTS: Visually, the graft appeared normal and incorporated with the native tendon. No repeated tear was observed, and results of tests for infection were negative. Histologically, the graft was infiltrated predominantly with fibroblasts and demonstrated numerous blood vessels. Positive proteoglycan staining in the AHDM and at sites of vascularity indicated probable transformation to tendon-like tissue. CONCLUSIONS: These histologic findings suggest that the AHDM is highly biocompatible, supports revascularization and repopulation with noninflammatory host cells, and becomes incorporated by surrounding tendon tissue. PMID- 19299346 TI - Treatment of plantar fasciitis using four different local injection modalities: a randomized prospective clinical trial. AB - BACKGROUND: To determine the effectiveness of four different local injection modalities in the treatment of plantar fasciitis. METHODS: In a prospective randomized multicenter study of plantar fasciitis, 100 patients were divided into four equal groups and were treated using four different methods of local injection: group A was treated with 2 mL of autologous blood alone; group B, an anesthetic (2 mL of lidocaine) combined with peppering; group C, a corticosteroid (2 mL of triamcinolone) alone; and group D, a corticosteroid (2 mL of triamcinolone) combined with peppering. The outcome was defined by using a 10-cm visual analog scale and modified criteria of the Roles and Maudsley score 3 weeks and 6 months after the injection and compared with the pretreatment condition. RESULTS: The successful results in all of the groups after injections were higher than those in the pretreatment condition (P = .000). In groups C and D, in which local corticosteroid injections were used, excellent results were obtained, with superior effect in the group in which peppering was used (P < .05). CONCLUSIONS: In the treatment of plantar fasciitis, combined corticosteroid injections and peppering is effective and produces better clinical results. PMID- 19299347 TI - Preclinical symptoms of the diabetic foot. AB - BACKGROUND: The diabetic foot is one of the main complications of diabetes mellitus, with a high risk of minor or major amputation. The preclinical foot lesions of patients without foot complaints were compared with healthy controls and analyzed. METHODS: This study was conducted with 89 diabetic patients from an endocrinology clinic and 35 nondiabetic control patients. The patients were asked about the presence, types, and durations of pedal complaints; acquired and congenital foot deformities; and atrophy. Patient gaits were inspected for any swelling; skin and nail changes were also recorded. Ranges of articular motion, deformities, crepitations, and any painful perceptions were noted. RESULTS: The differences between groups were significant for sensorial defects, joint changes of the foot, nail abnormalities, and neuropathic changes. CONCLUSIONS: Every patient with an established diagnosis of diabetes can be considered a potential sufferer of diabetic foot for whom medical therapy and foot protection programs are indicated. PMID- 19299348 TI - Knowledge, beliefs, and use of complementary and alternative medicine by Australian podiatric physicians. AB - BACKGROUND: Complementary and alternative medicine (CAM) is one of the fastest growing areas of health care. This has necessitated an increased awareness and understanding of CAM by conventional health professionals. METHODS: A questionnaire seeking information about use of and attitudes toward CAM was mailed to 1,365 Australian podiatric physicians. RESULTS: Ninety-one percent of Australian podiatric physicians surveyed have used at least one CAM therapy in the past 12 months, and 93% have treated patients with CAM or have recommended its use to patients. Overall, the respondents rated their knowledge of various CAM therapies as "average," and responses on the CAM Health Belief Questionnaire indicated that respondents tended not to endorse CAM health beliefs, with statements about CAM therapies being seen as "a threat to public safety" and effects being "usually due to the placebo effect" producing the strongest responses. CONCLUSIONS: Complementary and alternative medicine therapies are already being used in podiatric medical practice, and there are significant opportunities for further research into CAM education and clinical research relevant to podiatric medicine. PMID- 19299349 TI - Flexor tendon sheath as a source of pain in lesser metatarsal overload. AB - BACKGROUND: Lesser metatarsal overload may lead to pathologic conditions ranging from plantar metatarsophalangeal joint pain to dislocation. The flexor tendon sheath lies directly plantar to the joint. The increased pressure associated with lesser metatarsal overload may lead to synovitis and pain of the flexor tendon sheath. METHODS: Fifteen consecutive patients with lesser metatarsal overload had visual analog scale scores determined at three metatarsophalangeal joint areas to determine the source of pain. The patients underwent seven maneuvers to determine the pain scale score: single-leg heel raise and palpation of the second and third metatarsophalangeal joints on the plantar proximal, plantar distal, and dorsal aspects. The patients were then injected with 0.5 mL of lidocaine, 2%, into the second and third flexor tendon sheaths, and the maneuvers were repeated. RESULTS: Before the injection, the second metatarsophalangeal joint had a significantly greater visual analog scale score plantar than dorsal (6.9 and 2.6, P < or = .01). The flexor tendon sheath injection significantly improved all seven pain scale scores. CONCLUSIONS: Patients with lesser metatarsal overload experienced significantly greater visual analog scale pain scores on the plantar than the dorsal aspect of the second metatarsophalangeal joint. The scores significantly improved after diagnostic injection in the flexor tendon sheaths. The flexor tendon sheaths are probably involved in patients with primarily plantar pain. PMID- 19299350 TI - Diagnosis and prevalence of onychomycosis in diabetic neuropathic patients: an observational study. AB - BACKGROUND: An observational study was conducted to assess the prevalence of onychomycosis in clinically suspected diabetic neuropathic patients and to assess the reliability of the diagnosis. METHODS: One hundred successive type 1 and 2 diabetic patients with diabetic neuropathy were followed. Diabetic neuropathy was defined by a vibration perception threshold greater than 25 V and onychomycosis by clinical diagnosis. Samples of the most affected nail were taken. Potassium hydroxide testing and culture were performed. Photographs of the nails were used by two dermatologists for diagnosis. RESULTS: The mean +/- SE age was 62.3 +/- 11.4 years for the 20 onychomycotic patients and 60.3 +/- 10.4 years for the entire cohort; 14 onychomycotic patients (70%) were male versus 56 in the full cohort (56%) (P < .05). The prevalence of onychomycosis was 20% (culture and potassium hydroxide test positive) and 24% (culture positive). Twenty or 30 patients were positive by the potassium hydroxide test, depending on the investigator. The most frequent pathogen found was Trichophyton rubrum (11 of 20 patients; 55%). The positive predictive values of the dermatologist's diagnoses were 57.8% and 35.6%, and the negative predictive values were 85.0% and 90.5%. The two expert's results were significantly different (P < .05). CONCLUSIONS: The diagnosis of onychomycosis is difficult to make. The diagnostic methods commonly used are not satisfactory. If onychomycosis is dangerous for the diabetic foot, a better diagnostic method is needed. PMID- 19299351 TI - Psoriatic onychopachydermoperiostitis (POPP): a perplexing case study. AB - The following case of a 37-year-old male describes a very rare form of psoriasis known as psoriatic onychopachydermoperiostitis. There have been fewer than 20 cases reported worldwide. PMID- 19299352 TI - Varicosities as an etiology of tarsal tunnel syndrome and the significance of tinel's sign: report of two cases in young men and a review of the literature. AB - Two cases of tarsal tunnel syndrome in two young men with surgical and radiologic details are reported. The role of varicosities as a cause of tarsal tunnel syndrome and the significance of Tinel's sign are discussed through a large review of the literature. PMID- 19299353 TI - Plantar epidermoid inclusion cyst as a possible cause of intractable plantar keratosis lesions. AB - Inclusion cysts are benign lesions that appear as a consequence of traumatic inclusion of epidermal cells into the dermis. They can be painful if they appear under pressure areas, especially the metatarsal heads. We report a case of a 36 year-old woman with an intractable plantar keratosis lesion under the third metatarsal head of 3 years' duration. Ultrasonography revealed the presence of a subcutaneous mass with a growing epidermoid. It was surgically excised, and pathology confirmed the diagnosis of a plantar epidermoid cyst. All symptoms disappeared after the excision of the lesion. This case should alert the clinician about the existence of keratotic lesions in the metatarsal heads commonly diagnosed and treated as intractable plantar keratosis, although they are not directly derived from metatarsal overload. Some of the lesions could be directly derived from skin problems aggravated by pressure from the metatarsal head. This should be taken into consideration when addressing the management of these lesions. PMID- 19299354 TI - Lipoma arborescens of the peroneus longus and peroneus brevis tendon sheath: case report. AB - Lipoma arborescens is an uncommon pseudotumoral synovial lesion usually located in the suprapatellar pouch of the knee. Lipoma arborescens involving the synovial sheaths of the tendons is exceedingly rare. This diagnosis should be considered, particularly in patients with chronic joint effusion. We report a case with lipoma arborescens affecting the synovial sheaths of the peroneal tendons without involvement of the adjacent ankle joint. To our knowledge, this is the second reported case of lipoma arborescens involving tenosynovial sheaths of tendons around the ankle joint without ankle joint involvment. PMID- 19299355 TI - An enlarging distal tibia osteochondroma in the adult patient. AB - The extensive enlargement of a solitary osteochondroma in a skeletally mature patient is rare and might result from malignant transformation. Excision of such a lesion in the distal and lateral aspect of the tibia is difficult because of the risk of injury to the neurovascular structures and the possible functional consequences with respect to ankle stability. We present a case of an active osteochondroma arising from the posterolateral distal tibia in an adult patient. The tumor was successfully excised by using a transfibular approach with fibular reconstruction. No signs of recurrence were noted 2 years after surgery. PMID- 19299356 TI - Treatment of hallux valgus with three-dimensional modification of Mitchell's osteotomy: technique and results. AB - Mitchell's osteotomy gives very good results but there are still some cases where the original method, as well as its modification, cannot address all aspects of deformity. We modified the original Mitchell's method to address pronation and plantar displacement of the first metatarsal. Modification includes formation of lateral and plantar spur with metatarsal displacement and derotation of distal metatarsal fragment in the frontal and horizontal planes with stable screw fixation. We present midterm results of the first 60 patients compared to the original Mitchell method (30 patients). Differences between the groups postoperatively were in declination angle, postoperative metatarsalgia rate, and first metatarsal pronation angle. The technique described eliminated many of the disadvantages of Mitchell's method. PMID- 19299357 TI - Can pimecrolimus enhance wound healing? PMID- 19299358 TI - Poor sitting posture and metatarsus adductus deformity. PMID- 19299359 TI - Letter to the editor: the flat-footed child-to treat or not to treat what is the clinician to do? PMID- 19299361 TI - Inhaled anesthetics present cost-saving opportunity. PMID- 19299362 TI - New gout treatment approved. PMID- 19299363 TI - Experts sound warning on drug-resistant tuberculosis. PMID- 19299364 TI - FDA panel debates adding a fourth strain to influenza vaccine. PMID- 19299365 TI - Drug shortage broaches ethics of buying in excess. PMID- 19299366 TI - Remote management of medication orders. PMID- 19299368 TI - Pharmacist leads primary care team to improve diabetes care. PMID- 19299369 TI - Pharmacogenetics: from discovery to patient care. AB - PURPOSE: The basic concepts of pharmacogenetics, pharmacogenetic study approaches, factors to consider when applying pharmacogenetic discoveries to patient care, and potential roles for pharmacists in pharmacogenetics are discussed. SUMMARY: The Food and Drug Administration (FDA) has recognized pharmacogenomics as an opportunity to identify new biomarkers that may expedite the drug development process. Currently, there are over 50 drugs with pharmacogenetic discoveries on their labeling. Sequence variations in drug disposition genes can alter the pharmacokinetics of a drug, while sequence variations in drug target genes can change the pharmacodynamics of the drug. The two most common strategies to test a pharmacogenetic question are the candidate gene approach and genomewide association study. Given the complex interplay among the many factors that influence a drug dose, determination of an appropriate dose of a particular drug for a given patient will eventually require knowledge about both genetic and nongenetic factors that affect drug disposition and pharmacodynamics. Many factors can influence the application of pharmacogenetic discoveries to patient care. Before these discoveries find widespread application in clinical practice, additional work is needed, including randomized clinical trials to evaluate the clinical utility of a pharmacogenetic test, the development of guidelines for the clinical use of various pharmacogenetic tests, and provider education on pharmacogenetics. CONCLUSION: Pharmacogenetics has made significant progress in the past decade, and many pharmacogenetic discoveries have now been included on FDA-approved drug labeling. Pharmacogenetic discoveries may further promote safe and effective use of medications by more accurately predicting an individual's drug response. PMID- 19299370 TI - Probable enoxaparin-induced hepatotoxicity. AB - PURPOSE: A case of probable enoxaparin-induced hepatotoxicity is described. SUMMARY: A 29-year-old woman sought treatment from a pulmonologist for a dry, hacking, constant cough not relieved by fast-acting inhalers or narcotic cough medications that had lasted for three weeks. Her primary care physician had earlier made a preliminary diagnosis of pertussis and prescribed a short course of azithromycin and corticosteroids, which did not help relieve the symptoms. Computed tomography angiography of her chest revealed multiple bilateral pulmonary emboli with a moderate clot burden, which resulted in her hospitalization. The pulmonary emboli were thought to be associated with her oral contraceptive, which was discontinued at hospital admission. Anticoagulant therapy was initiated with subcutaneous enoxaparin and oral warfarin. Beginning the second day of therapy, the patient complained of nausea and associated vomiting. Diagnostic procedures did not reveal any liver, kidney, splenic, or pancreatic abnormalities. The results of laboratory tests revealed elevated levels of hepatic enzymes, including alanine transaminase (ALT) and aspartate transaminase (AST). Tests for hepatitis A, B, and C were negative. Enoxaparin therapy was discontinued, and the patient was maintained on oral warfarin. Clinical and laboratory signs of liver injury resolved over the next few days, with a return to baseline levels of AST and ALT levels over the subsequent months. According to the Naranjo et al. adverse-reaction probability scale, enoxaparin was the probable cause of hepatotoxicity in this patient. CONCLUSION: A woman receiving enoxaparin every 12 hours developed signs and symptoms of hepatotoxicity after the second dose. The case was unusual in the rapidity and magnitude of hepatic enzyme elevation. PMID- 19299371 TI - Estimation of creatinine clearance in morbidly obese patients. AB - PURPOSE: Estimates of creatinine clearance (CL(cr)) based on equations and various body-size descriptors were compared with 24-hour measured CL(cr) values in morbidly obese patients. METHODS: Patients age 18-75 years with a body mass index (BMI) of >/=40 kg/m(2) with stable serum creatinine values were enrolled. Covariates known to contribute to alteration in CL(cr) were used to exclude patients. Twenty-four-hour urine collection was performed to measure CL(cr). Bioelectric impedance analysis was used to estimate fat-free weight (FFW). Glomerular filtration rate was estimated using the four-variable Modification of Diet in Renal Disease (MDRD4) equation. CL(cr) was estimated using the Cockcroft Gault and Salazar-Corcoran methods using total body weight (TBW). Body-size descriptors, such as ideal body weight (IBW), adjusted body weight (ABW), and lean body weight (LBW), and FFW were substituted in the Cockcroft-Gault equation to generate additional estimates of CL(cr). RESULTS: Fifty-four patients (mean +/ S.D. age, 48.4 +/- 12.9 years; TBW, 142.3 +/- 41.7 kg; BMI, 50.5 +/- 12.6 kg/m(2)) completed the study. All three equations were biased in their estimation of CL(cr). Use of MDRD4 and IBW in the Cockcroft-Gault equation underestimated CL(cr), while the Salazar-Corcoran equation and use of TBW or ABW in the Cockcroft-Gault equation overestimated this value. Substitution of fat-free weight or LBW in the Cockcroft-Gault equation provided unbiased estimates of CL(cr). CONCLUSION: An LBW estimate, based on TBW and BMI, incorporated into the Cockcroft-Gault equation provided an unbiased, relatively precise, accurate, and clinically practical estimate of 24-hour measured CL(cr) in morbidly obese patients. PMID- 19299372 TI - Lipid globule size in total nutrient admixtures prepared in three-chamber plastic bags. AB - PURPOSE: The stability of injectable lipid emulsions in three-chamber plastic (3CP) bags, applying the globule-size limits established by United States Pharmacopeia ( USP ) chapter 729, was studied. METHODS: A total of five premixed total nutrient admixture (TNA) products packaged in 3CP bags from two different lipid manufacturers containing either 20% soybean oil or a mixture of soybean oil and medium-chain-triglyceride oil as injectable lipid emulsions were tested. Two low-osmolarity 3CP bags and three high-osmolarity 3CP bags were studied. All products were tested with the addition of trace elements and multivitamins. All additive conditions (with and without electrolytes) were tested in triplicate at time 0 (immediately after mixing) and at 6, 24, 30, and 48 hours after mixing; the bags were stored at 24-26 degrees C. All additives were equally distributed in each bag for comparative testing, applying both globule sizing methods outlined in USP chapter 729. RESULTS: Of the bags tested, all bags from one manufacturer were coarse emulsions, showing signs of significant growth in the large-diameter tail when mixed as a TNA formulation and failing the limits set by method II of USP chapter 729 from the outset and throughout the study, while the bags from the other manufacturer were fine emulsions and met these limits. Of the bags that failed, significant instability was noted in one series containing additional electrolytes. CONCLUSION: Injectable lipid emulsions provided in 3CP bags that did not meet the globule-size limits of USP chapter 729 produced coarser TNA formulations than emulsions that met the USP limits. PMID- 19299373 TI - Relationships between beliefs about medications and adherence. AB - PURPOSE: The relationships between beliefs about medications, health literacy, and self-reported medication adherence are examined. METHODS: Patients from an inner-city hospital pharmacy completed an in-person, interviewer-assisted questionnaire that included the Morisky 8-item Medication Adherence Scale (MMAS 8), the Beliefs About Medicines Questionnaire (BMQ), and the Rapid Estimate of Adult Literacy in Medicine (REALM). Multivariable logistic regression was used to determine predictors of self-reported medication adherence as determined by the MMAS-8. Variables included in the model were summary scores from the BMQ, REALM, and patient or regimen characteristics that were significantly associated with the MMAS-8. RESULTS: A majority of the 275 study participants were African American (86.2%), were women (73.1%), and could read at less than a high school reading level (59.7%). The average age was 53.9 years. Approximately half of the patients (52.7%) reported low medication adherence (MMAS-8 score of >2). Multivariate analyses indicated several factors were associated with low self reported adherence, including negative beliefs about medications, younger age, low medication self-efficacy, and hyperlipidemia. Health literacy was not independently associated with beliefs or adherence. CONCLUSION: Patients who had negative beliefs about medications, who were <65 years of age, or who had low medication self-efficacy reported low medication adherence. PMID- 19299374 TI - Stability of extemporaneously prepared moxifloxacin oral suspensions. AB - PURPOSE: The stability of extemporaneously prepared moxifloxacin oral suspensions was studied. METHODS: An oral suspension of moxifloxacin 20 mg/mL was prepared by thoroughly grinding three 400-mg tablets of moxifloxacin in a glass mortar. Thirty milliliters of Ora-Plus and 30 mL of either Ora-Sweet or Ora-Sweet SF were mixed and added to the powder to make a final volume of 60 mL. Three identical samples of each formulation were prepared and placed in 2-oz amber plastic bottles with child-resistant caps and were stored at room temperature (23-25 degrees C). A 1-mL sample was withdrawn from each of the six bottles with a micropipette immediately after preparation and at 7, 14, 28, 60, and 90 days. After further dilution to an expected concentration of 8 microg/ mL with sample diluent, the samples were assayed in duplicate by stability-indicating high performance liquid chromatography. Stability was defined as the retention of at least 90% of the initial concentration. RESULTS: At least 99% of the initial moxifloxacin remained throughout the 90-day study period in both preparations. There were no detectable changes in color, odor, taste, and pH and no visible microbial growth in any sample. CONCLUSION: Extemporaneously compounded suspensions of moxifloxacin 20 mg/mL in a 1:1 mixture of Ora-Plus and Ora-Sweet or Ora-Sweet SF were stable for at least 90 days when stored in 2-oz amber plastic bottles at room temperature. PMID- 19299375 TI - Opportunity cost of pharmacists' nearly universal prospective order review. PMID- 19299376 TI - Maintaining the enterprisewide continuity and interoperability of patient allergy data. AB - PURPOSE: The efforts, results, and challenges of a large tertiary care, academic health care system to standardize and integrate allergy information across clinical information systems are discussed. SUMMARY: The University of Michigan Health System and its Information Technology Strategic Advisory Committee recognized the necessity for storing and maintaining allergy information in a single repository; therefore, the clinical data repository (CDR) was named as the central database for coded allergens and reactions for the University of Michigan Hospitals and Health Care Centers (UMHHC) electronic medical record. The Enterprise Allergy Project (EAP) began in June 2005 with the formation of a steering committee that included representatives from clinical departments with order-entry systems. The initial phase of the EAP consisted of several components. One component was a one-time conversion of existing free-text allergy information into coded allergens and reactions. Before the implementation of the EAP, the order-entry system only supported the entry of uncoded allergen and reaction information. An initial process of allergy matching reduced the list of un-coded allergens from 272,519 to 29,500 by using terms that indicated no allergies were present and trimming and modifying free-text strings that closely matched or easily translated to a coded allergen counterpart. Another component of the EAP consisted of the interface and technical build to support allergy information processing between the CDR and University of Michigan (UM)-Carelink. One goal of the EAP was to transfer data bidirectionally, but that goal could not safely be accomplished. CONCLUSION: Implementing a strategy for enterprise allergy integration at UMHHC has improved the quality of allergy information documented as measured by a significant decrease in the amount of uncoded allergens. PMID- 19299377 TI - Hospital liability for accuracy of pharmacist consultations. PMID- 19299378 TI - Recent advances in coeliac disease genetics. PMID- 19299379 TI - Balancing the risks and benefits of prolonged use of infliximab. PMID- 19299380 TI - The true culprit within the SPINK1 p.N34S-containing haplotype is still at large. PMID- 19299381 TI - A fat chance for hepatocyte transplantation? PMID- 19299382 TI - Bloody diarrhoea in a 54-year-old Arab patient. PMID- 19299383 TI - Does old age cause abdominal pain? PMID- 19299384 TI - "Odd-looking" oesophageal varices. PMID- 19299386 TI - Right upper quadrant pain and fever following a fall in a 92 year old. PMID- 19299387 TI - Liver failure complicating segmental hepatic ischaemia induced by a PTFE-coated TIPS stent. AB - The use of polytetrafluoroethylene (PTFE)-covered prostheses improves trans jugular intrahepatic porto-systemic shunt (TIPS) patency and decreases the incidence of clinical relapses and re-interventions. Therefore, the improvement provided by covered stents might expand the currently accepted recommendations for TIPS use. Stent-related occlusion of the hepatic vein with consequent ischaemia of the corresponding liver parenchyma emerges as a novel complication reported in at least 5% of patients implanted with coated stents. However, this complication was reported to be mild, without signs or symptoms of liver failure, and self-limiting. We report a case of segmental liver ischaemia following PTFE covered stent placement resulting in a marked impairment in liver function in a patient with hepatitis C virus cirrhosis implanted because of refractory oesophageal bleeding, thus expanding the severity range of this new procedural complication. Moreover, we discuss the possible involvement of additional pathogenetic mechanisms other than out-flow obstruction in the onset of coated stent induced congestive liver ischaemia. PMID- 19299388 TI - Narrow-band imaging for improving colorectal adenoma detection: appropriate system function settings are required. PMID- 19299389 TI - The utility of using bispectral index monitoring as an early intraoperative indicator of initial poor graft function after orthotopic or split-graft liver transplantation. PMID- 19299390 TI - Progressive multifocal leukoencephalopathy in autoimmune disorders. PMID- 19299391 TI - Pregnancy: a risk factor for Budd-Chiari syndrome? PMID- 19299392 TI - Statistics, damned statistics and time to intervene. PMID- 19299393 TI - Certolizumab use in pregnancy. PMID- 19299395 TI - Germline variants of IRGM in childhood-onset Crohn's disease. PMID- 19299397 TI - The Obama presidency: what may happen, what needs to happen in health policies in the USA. PMID- 19299398 TI - Challenges in measuring changes in health and social indicators over time. PMID- 19299399 TI - Taking the reins of the white horse of climate change. PMID- 19299400 TI - Retrofitting houses with insulation: a cost-benefit analysis of a randomised community trial. AB - BACKGROUND: Housing is an important environmental influence on population health, and there is growing evidence of health effects from indoor environment characteristics such as low indoor temperatures. However, there is relatively little research, and thus little firm guidance, on the cost-effectiveness of public policies to retrospectively improve the standards of houses. The purpose of this study was to value the health, energy and environmental benefits of retrofitting insulation, through assessing a number of forms of possible benefit: a reduced number of visits to GPs, hospitalisations, days off school, days off work, energy savings and CO(2) savings. METHODS: All these metrics are used in a cluster randomised trial--the "Housing, Insulation and Health Study"--of retrofitting insulation in 1350 houses, in which at least one person had symptoms of respiratory disease, in predominantly low-income communities in New Zealand. RESULTS: Valuing the health gains, and energy and CO(2) emissions savings, suggests that total benefits in "present value" (discounted) terms are one and a half to two times the magnitude of the cost of retrofitting insulation. CONCLUSION: This study points to the need to consider as wide a range of benefits as possible, including health and environmental benefits, when assessing the value for money of an intervention to improve housing quality. From an environmental, energy and health perspective, the value for money of improving housing quality by retrofitting insulation is compelling. PMID- 19299401 TI - How political epidemiology research can address why the millennium development goals have not been achieved: developing a research agenda. AB - The Millennium Development Goals (MDG) progress targets have not been met. Nevertheless, the United Nations (UN) has not yet undertaken in-depth review in order to discover the reasons behind this lack of progress in achieving the MDG. From a political epidemiology perspective, the intention here is to identify the political elements affecting the social factors impeding MDG fulfilment and, at the same time, to suggest future public policies and appropriate proposals that are both more coherent and supported by broader, empirical knowledge of the relevant issues. PMID- 19299402 TI - Is house-dust nicotine a good surrogate for household smoking? AB - The literature is inconsistent regarding associations between parental smoking and childhood leukemia, possibly because previous studies used self-reported smoking habits as surrogates for children's true exposures to cigarette smoke. Here, the authors investigated the use of nicotine concentrations in house dust as measures of children's exposure to cigarette smoke in 469 households from the Northern California Childhood Leukemia Study (1999-2007). House dust was collected by using high-volume surface samplers and household vacuum cleaners and was analyzed for nicotine via gas chromatography-mass spectrometry. Using multivariable linear regression, the authors evaluated the effects of self reported parental smoking, parental demographics, house characteristics, and other covariates on house-dust nicotine concentrations. They observed that nicotine concentrations in house dust were associated with self-reported smoking for periods of months and years before dust collection. Furthermore, the authors found that the relation between nicotine dust levels and self-reported smoking varied by parental age and socioeconomic status. These findings suggest that house-dust nicotine concentrations reflect long-term exposures to cigarette smoke in the home and that they may be less biased surrogates for children's exposures to cigarette smoke than self-reported smoking habits. PMID- 19299403 TI - Alcohol use and risk of pancreatic cancer: the NIH-AARP Diet and Health Study. AB - The epidemiologic evidence for the role of alcohol use in pancreatic cancer development is equivocal. The authors prospectively examined the relation between alcohol use and risk of pancreatic cancer among 470,681 participants who were aged 50-71 years in 1995-1996 in the US National Institutes of Health-AARP Diet and Health Study. The authors identified 1,149 eligible exocrine pancreatic cancer cases through December 2003. Multivariate Cox proportional hazards regression models were used to calculate relative risks and 95% confidence intervals with the referent group being light drinkers (<1 drink/day). The relative risks of developing pancreatic cancer were 1.45 (95% confidence interval (CI): 1.17, 1.80; P(trend) = 0.002) for heavy total alcohol use (>or=3 drinks/day, approximately 40 g of alcohol/day) and 1.62 (95% CI: 1.24, 2.10; P(trend) = 0.001) for heavy liquor use, compared with the respective referent group. The increased risk with heavy total alcohol use was seen in never smokers (relative risk = 1.35, 95% CI: 0.79, 2.30) and participants who quit smoking 10 or more years ago before baseline (relative risk = 1.41, 95% CI: 1.01, 2.00). These findings suggest a moderately increased pancreatic cancer risk with heavy alcohol use, particularly liquor; however, residual confounding by cigarette smoking cannot be completely excluded. PMID- 19299404 TI - Plasma urate and Parkinson's disease in the Atherosclerosis Risk in Communities (ARIC) study. AB - Higher plasma urate concentration has been linked to lower risk of Parkinson's disease in men, but data are lacking on women and African Americans. The authors examined plasma urate in relation to Parkinson's disease in the biracial, population-based Atherosclerosis Risk in Communities (ARIC) cohort. Between 1987 and 1989, 15,792 participants, aged 45-64 years, were recruited from 4 US communities and have since been followed with 3 triennial visits and annual surveillance. Plasma urate was measured at visits 1 and 2, and the concentrations were highly correlated. From visit 1 through 2004, 95 potential cases of Parkinson's disease were identified from multiple sources. Odds ratios and 95% confidence intervals were calculated from multivariate logistic regression models. Plasma urate concentration was inversely associated with Parkinson's disease occurrence. The odds ratios between extreme quartiles of plasma urate were 0.4 (95% confidence interval: 0.2, 0.8) in the overall analysis, 0.3 (95% confidence interval: 0.1, 0.7) for men, and 0.4 (95% confidence interval: 0.2, 1.0) for Caucasians. Such an association was also suggested among women and African Americans but was not statistically significant because of small sample sizes. These data support the previous finding that urate may be a protective factor against Parkinson's disease. PMID- 19299405 TI - Do socioeconomic gradients in body mass index vary by race/ethnicity, gender, and birthplace? AB - Despite the well-documented negative socioeconomic status (SES) gradient in body mass index (BMI; weight (kg)/height (m)(2)) among women in developed societies, the presence and strength of the gradient is less consistent among men. Far less clear is the SES patterning of BMI among racial/ethnic minorities and immigrants. Using data from the 2001 California Health Interview Survey, a cross-sectional representative sample of California adults, the authors examined whether the SES patterning of BMI varied across 4 major US racial/ethnic groups (n = 37,150) by gender and birthplace. The shape and strength of the relation between SES and BMI differed markedly by race/ethnicity; and within racial/ethnic groups, it varied by gender. Irrespective of race/ethnicity, there were negative income and education gradients in BMI among women; however, there was considerable variation among men. The effect of education on BMI differed by birthplace in some groups. A clear education gradient in BMI was found among all US-born participants, a quadratic education pattern in BMI was found among foreign-born Asian men, a flat pattern was found among foreign-born Asian women, and no clear pattern was found in the remaining foreign-born groups. There is substantial heterogeneity in the contemporaneous SES gradient in BMI. US social disparities in BMI require simultaneous consideration of race/ethnicity and SES, but also birthplace. PMID- 19299406 TI - Sleep duration in the United States: a cross-sectional population-based study. AB - Sleep duration is associated with cardiovascular disease and diabetes risk factors, depression, automobile and workplace accidents, and prospective mortality. Little is known, however, about sleep patterns in the US population. The 2004-2007 National Health Interview Survey-Sample Adult Files provide nationally representative data for 110,441 noninstitutionalized US adults aged 18 years or older, and multinomial logistic regression examines whether variables in 5 domains-demographic, family structure, socioeconomic, health behavior, and health status-are associated with long or short sleep duration. Being older, non Hispanic black, or a current or former smoker; having low levels of education, income, or few income sources; consuming few or numerous drinks in a week; or reporting cardiovascular disease, diabetes, depression, underweight, or activity limitations is associated with increased odds of both long and short sleep duration. Other variables are associated with shorter (e.g., living with young children, being unmarried, working long hours, more frequent binge drinking) or longer (e.g., being younger, Mexican American, pregnant, or having low levels of physical activity) sleep hours. The authors identify numerous risk factors for long and short sleep; many of those variables are potential confounders of the relation between sleep hours and other health outcomes. PMID- 19299407 TI - Replication of genetic associations with plasma lipoprotein traits in a multiethnic sample. AB - Recent genome-wide association studies (GWAS) have reproducibly identified loci associated with plasma triglycerides (TG), HDL cholesterol, and LDL cholesterol. We sought to replicate these findings in a multiethnic population-based cohort using the curated single nucleotide polymorphism (SNP) set found on the new Illumina cardiovascular disease (CVD) beadchip, which contains approximately 50,000 SNPs densely mapping approximately 2,100 genes, selected based on their potential role in CVD. The sample consisted of individuals with European (n = 272), South Asian (n = 330), and Chinese (n = 304) ancestry. Identity by state clustering successfully classified individuals according to self-reported ethnicities. Associations between TG and APOA5, TG and LPL, HDL and CETP, and LDL and APOE were all identified (P < 2 x 10(-6)). In 13 loci, associations with the same SNP or a proxy SNP were identified in the same direction as previously reported (P < 0.05). Assessing the cumulative number of risk-associated alleles at multiple replicated SNPs increased the proportion of explained lipoprotein variance over and above traditional variables such as age, sex, body mass index, and ethnicity. The findings indicate the potential utility of the Illumina CVD beadchip, but they underscore the need to consider meta-analysis of results from commonly studied clinical or epidemiological samples. PMID- 19299408 TI - Does confirmatory tumor biopsy alter the management of breast cancer patients with distant metastases? AB - BACKGROUND: Decisions about systemic treatment of women with metastatic breast cancer are often based on estrogen receptor (ER), progesterone receptor (PgR), and Her2 status of the primary tumor. This study prospectively investigated concordance in receptor status between primary tumor and distant metastases and assessed the impact of any discordance on patient management. MATERIALS AND METHODS: Biopsies of suspected metastatic lesions were obtained from patients and analyzed for ER/PgR and Her2. Receptor status was compared for metastases and primary tumors. Questionnaires were completed by the oncologist before and after biopsy to determine whether the biopsy results changed the treatment plan. RESULTS: Forty women were enrolled; 35 of them underwent biopsy, yielding 29 samples sufficient for analysis; 3/29 biopsies (10%) showed benign disease. Changes in hormone receptor status were observed in 40% (P = 0.003) and in Her2 status in 8% of women. Biopsy results led to a change of management in 20% of patients (P = 0.002). CONCLUSIONS: This prospective study demonstrates the presence of substantial discordance in receptor status between primary tumor and metastases, which led to altered management in 20% of cases. Tissue confirmation should be considered in patients with clinical or radiological suspicion of metastatic recurrence. PMID- 19299409 TI - Immediate and short-term impact of a brief motivational smoking intervention using a biomedical risk assessment: the Get PHIT trial. AB - INTRODUCTION: Providing smokers with biologically based evidence of smoking related disease risk or physical impairment may be an effective way to motivate cessation. METHODS: Smokers were recruited for a free health risk assessment and randomized to receive personally tailored feedback based on their lung functioning, carbon monoxide (CO) exposure, and smoking-related health conditions or generic information about the risks of smoking and personalized counseling based on their diet, body mass index, and physical activity. All (n = 536) were advised to quit smoking and offered access to a free telephone cessation program. Participants were surveyed immediately after intervention and 1 month later to assess the impact on various indices of motivation to quit. RESULTS: Immediately posttreatment, experimental participants rated themselves as more likely to try to quit (p = .02) and reported a greater mean increase in their motivation to quit than controls (p = .04). At 1-month follow-up, however, we found no significant group differences on any motivational indices. In post-hoc analyses comparing smokers in the experimental group with and without lung impairment, persons with impaired lung functioning had a greater change from baseline in posttreatment motivation to quit (adjusted p = .05) and perceived risk of developing a smoking-related disease (p = .03) compared with persons with no lung impairment, but we found no significant treatment effect on any motivational indices at 1 month. DISCUSSION: The results suggest that the intervention had a small, temporary effect, but we found no clear evidence that the intervention increased motivation to quit smoking during the first month postintervention. PMID- 19299410 TI - Smoking cessation behavior in older adults by race and gender: the role of health problems and psychological distress. AB - INTRODUCTION: Initial research on older smokers suggests that a subgroup of smokers with higher levels of psychological distress and health problems may be more likely to quit smoking than older smokers with fewer such problems. The present study, based on prospective data from a biracial sample of older adults (N = 4,162), examined characteristics of older adult smokers by race and gender. METHODS: The present study uses both cross-sectional and prospective data to examine the association between smoking behavior, smoking cessation, health functioning, and psychological distress in a biracial sample of community dwelling older adults. RESULTS: We found baseline psychological distress to be associated with poor health functioning. Consistent with hypotheses, baseline (Time 1) psychological distress predicted smoking cessation 3 years later (Time 2). Moreover, the change in health problems between Time 1 and Time 2 fully mediated the association between Time 1 distress and smoking cessation. DISCUSSION: Smoking cessation behavior of older adults is best explained by higher levels of distress and health problems regardless of race or gender. These findings may have important treatment implications regarding smoking cessation programs among older adults. Older adult smokers with higher levels of psychological distress and health problems may be more motivated to quit smoking than those with fewer such problems. These difficulties should be targeted within the context of the smoking cessation protocol. Also, we identified a subgroup of older smokers who are reporting fairly good health and lower levels of distress and who are less likely to quit smoking. Motivational methods may need to be developed to engage this group in smoking cessation treatment. PMID- 19299414 TI - Postural headache. PMID- 19299415 TI - Micropapillary-type invasion in low-grade mammary carcinomas. PMID- 19299416 TI - Promoting public health research and collaboration in the Asia-Pacific region. PMID- 19299417 TI - Global asymptotic properties of virus dynamics models with dose-dependent parasite reproduction and virulence and non-linear incidence rate. AB - We consider two models for the spread of an infection with a free-living infective stage, where parasite reproduction and virulence (parasite-induced mortality) depend on the parasite dose to which the host is exposed and are given by unspecified non-linear functions of the number of the free pathogen particles, and the incidence rate is non-linear. We study the impact of these non linearities with the focus on the global properties of these models. We consider a very general form of the non-linearities: we assume that the virulence and the parasite reproduction rates are given by unspecified non-linear functions of the number of the free pathogen particles and that the incidence rate is an unspecified function of the number of susceptible hosts and free pathogen particles; all these functions are constrained by a few biologically feasible conditions. We construct Lyapunov functions that enable us to find biologically realistic conditions which are sufficient to ensure existence and uniqueness of a globally asymptotically stable equilibrium state. Depending on the value of the basic reproduction number, this equilibrium state can be either positive, where parasite endemically persists, or infection free. PMID- 19299418 TI - Rapid sublethal toxicity assessment using bioluminescent Caenorhabditis elegans, a novel whole-animal metabolic biosensor. AB - Sublethal metabolic effects are informative toxicological end points. We used a rapid quantitative metabolic end point, bioluminescence of firefly luciferase expressing Caenorhabditis elegans, to assess effects of sublethal chronic exposure (19 h) to the oxidative stress agent and environmental pollutant cadmium (provided as chloride salt). Bioluminescence declined in a concentration dependent manner in the concentration range tested (0-30 microM Cd), with comparable sensitivity to reproduction and developmental assay end points (after 67 and 72 h, respectively). Cd concentrations that resulted in 20% reduction in bioluminescence (EC(20)) were 11.8-13.0 microM, whereas the reproduction EC(20) (67 h exposure) was 10.2 microM. At low concentrations of Cd (< or = 15 microM), the decline in bioluminescence reflected a drop in ATP levels. At Cd concentrations of 15-30 microM, decreased bioluminescence was attributable both to effects of Cd on ATP levels and decreased production of luciferase proteins, concomitant with a decline in protein levels. We show that whole-animal bioluminescence is a valid toxicological end point and a rapid and sensitive predictor of effects of Cd exposure on development and reproduction. This provides a platform for high-throughput sublethal screening and will potentially contribute to reduction of testing in higher animals. PMID- 19299419 TI - Standard and molecular NOAELs for rat testicular toxicity induced by flutamide. AB - An important step in the safety assessment of chemicals for humans is to determine the no observed adverse effect level (NOAEL) in toxicity studies conducted in animal models. With the increasing use of molecular tools in toxicity studies, a question often posed is how a NOAEL derived from molecular data compares to a NOAEL established using standard methods. The objective of the present study was to address this question when considering testicular toxicity. To do this, we assessed the effects of the reference antiandrogen flutamide on rat testes in a standard 28-day toxicity study using doses of 0.04-150 mg/kg/day. At necropsy, blood samples were collected for testosterone measurements. The testes were collected for histopathological assessment as well as for the evaluation of gene expression changes using quantitative PCR analyses. Results showed that increases in plasma testosterone level and Leydig cell hyperplasia were detected from 6 mg/kg/day. An alteration in the level of accumulation of a selection of genes was also detected from 6 mg/kg/day. This was the case for genes functionally associated with the testicular lesion, such as lipid metabolism and cell death/cell proliferation, as well as for genes not functionally associated with the lesion. Contrary to the misgivings, these data show that, using a standard 28-day toxicity study and a well-characterized adverse effect, the NOAEL based on transcript changes is similar to the NOAELs based on testosterone levels and histopathological examination. PMID- 19299420 TI - Identification of novel SHPS-1-associated proteins and their roles in regulation of insulin-like growth factor-dependent responses in vascular smooth muscle cells. AB - Tyrosine phosphatase non-receptor type substrate-1 (SHPS-1), a transmembrane protein, plays a vital role in cell migration and proliferation. Our previous studies have shown that insulin-like growth factor-I (IGF-I) stimulates SHPS-1 phosphorylation, leading to recruitment of SHP-2, c-Src, Shc, and Grb2.p85 to phosphorylated SHPS-1. Assembly of this signaling complex is required for optimal stimulation of both mitogen-activated protein and phosphatidylinositol 3-kinase pathways. The main aim of the present study was to identify novel proteins that interacted with the cytoplasmic domain of SHPS-1 (SHPS-1/CD) in response to IGF-I stimulation and define the role of these interactions in mediating specific biological functions. We performed a functional proteomic screening to identify SHPS-1 binding partners using combination of mRNA display and the tandem affinity purification-tag methods. Screening identified a number of proteins not previously known to interact with phosphorylated SHPS-1/CD. These novel SHPS-1 binding partners represent several functional categories including heat shock proteins, protein kinases and phosphatases, and proteins that regulate transcription or translation. In Vivo and in vitro studies suggested that most of the proteins bound to SHPS-1 via binding to one of the four SH2 domain containing proteins, SHP-2, CTK, SUPT6H, and STAT1, that directly bound to SHPS-1. Although the binding of most of these proteins to SHPS-1 was positively regulated by IGF I, a few were negatively regulated, suggesting differential regulation of protein complexes assembled on SHPS-1/CD in response to IGF-I. Further studies showed that truncation of SHPS-1/CD significantly impaired IGF-I-dependent AKT signal transduction and subsequent biological functions including cell survival, protein synthesis, protein aggregation, and prevention of apoptosis. The results emphasize the importance of formation of SHPS-1 signaling complex induced by IGF I and provide novel insights into our knowledge of the role of this molecular scaffold in regulation of IGF-I-stimulated signal transduction and biological actions. PMID- 19299422 TI - Cardiac output by arterial pulse contour: reliability under hemodynamic derangements. AB - Pulse contour methods (PCM) for the measurements of cardiac output (CO) are gaining popularity in intensive care settings but their reliability during hemodynamic instability has been questioned. Pressure-recording-analytical-method (PRAM) is a newly developed uncalibrated hemodynamic monitor and its capability in measuring CO during hemodynamic instability is still under investigation. Dobutamine (2.5 and 5 microg/kg/min), vasoconstriction (arginine-vasopressin 4, 8 and 16 IU/h), hemorrhage (-10%, -20%, -35%, and -50% of the theoretical volemia), and volume resuscitation were induced in eight swine. CO by means of thermodilution (CO(ThD)), transesophageal echocardiography (CO(TEE)) and PRAM (CO(PRAM)) were contemporarily registered. R(2), bias, and percentage error were used to compare the methods. Comparison between CO(PRAM) and CO(ThD) resulted in: r(2)=0.87; bias=-0.006 l/min; precision=+/-0.87 l/min; percentage error=22.8%. Comparison between CO(PRAM) and CO(TEE) resulted in: r(2)=0.85; bias=-0.007 l/min; precision=+/-0.86 l/min; percentage error=22%. Sub-group analysis revealed disagreement between methods only during the last two steps of hemorrhage: CO(PRAM) vs. CO(ThD): r(2)=0.67, bias=-0.37 l/min, precision=+/-1.04 l/min, limits of agreement=-1.39+0.66 l/min, and percentage error=45%; CO(PRAM) vs. CO(TEE): r(2)=0.38, bias=0.4 l/min, precision=+/-1.42 l/min, limits of agreement= 0.99+1.79 l/min, and percentage error=62%. PRAM resulted to be accurate in measuring CO during hemodynamic stability, tachycardia, and vasoconstriction. When volemia was reduced by >35%, disagreement between methods was observed. PMID- 19299421 TI - Excretory/secretory proteome of the adult developmental stage of human blood fluke, Schistosoma japonicum. AB - Schistosomes are the causative agents of schistosomiasis, one of the most prevalent and serious of the parasitic diseases that currently infects approximately 200 million people worldwide. Schistosome excretory/secretory (ES) proteins have been shown to play important roles in modulating mammalian host immune systems. In our current study, we performed a global proteomics identification of the ES proteins from adult worms of Schistosoma japonicum, one of the three major schistosome species. Our results unambiguously identified 101 proteins, including 53 putatively secreted proteins. By quantitative analysis, we revealed fatty acid-binding protein as a major constituent of the in vitro ES proteome. Strikingly the heat shock proteins HSP70s, HSP90, and HSP97 constituted the largest protein family in the ES proteome, implying a central role for these proteins in immunomodulation in the host-parasite relationship. Other important S. japonicum ES proteins included actins, 14-3-3, aminopeptidase, enolase, and glyceraldehyde-3-phosphate dehydrogenase, some of which have been considered as viable vaccine candidates and therapeutic targets. A comparison with previous studies suggests that 48.5% of S. japonicum ES proteins are common to other parasite ES products, indicating that the molecular mechanisms involved in evading the host immune response may be conserved across different parasites. Interestingly seven host proteins, including antimicrobial protein CAP18, immunoglobulins, and a complement component, were identified among in vitro S. japonicum ES products likely originating from the schistosome tegument or gut, indicating that host innate and acquired immune systems could defend against schistosome invasion. Our present study represents the first attempt at profiling S. japonicum ES proteins, provides an insight into host-parasite interactions, and establishes a resource for the development of diagnostic agents and vaccines for the control of schistosomiasis. PMID- 19299423 TI - Aortic valve thrombosis after implantation of temporary left ventricular assist device. AB - The use of assist devices for ventricular support after myocardial infarction with cardiogenic shock has become common practice. Thrombosis, bleeding, and infection are common complications. However, native valve thrombosis is a rare complication. We present a case of aortic valve thrombosis after implantation of a left ventricular assist device (LVAD) treated with thrombus removal at time of device exchange. PMID- 19299424 TI - Clinical application of an ultrasonic scalpel to divide pulmonary vessels based on laboratory evidence. AB - The Harmonic Ace ultrasonic scalpel (Ethicon Endo-Surgery, Inc, Cincinnati, Ohio), has been widely used in endoscopic surgery to divide systemic vessels, but not pulmonary vessels. We describe our initial clinical experience of using it for pulmonary vessel division. The Harmonic Ace was used to divide pulmonary vessels 5 mm or less in diameter, secured with a proximal single ligation, in 20 patients who underwent video-assisted major lung resection between September 2007 and April 2008. We also evaluated the sealing potential of this device in a pig model. We divided 43 pulmonary arteries and 13 pulmonary veins (PV) by the device. The diameter of the divided vessels ranged from 2 to 5 mm. Vascular sealing was successful in all except two early procedures: several subsegmental arteries were held at once, and vessels positioned near the distal end of the blade jaw bled intraoperatively. There was no postoperative bleeding. In the pig model, the bursting pressure of sealed pulmonary arteries (PA) was >75 mmHg. Pulmonary vessels can be safely divided using the Harmonic Ace with proximal single ligation unless multiple or large pulmonary vessels are held within the blade jaw. PMID- 19299425 TI - Eliminating neglected diseases in poor countries: a conversation with Andrew Witty. Interview by Susan Dentzer. PMID- 19299426 TI - Memorial. PMID- 19299427 TI - Effects of enalapril, candesartan or both on neurohumoral activation and LV volumes and function in patients with heart failure not treated with a beta blocker. AB - BACKGROUND: The long-term effects of the angiotensin-receptor blocker candesartan, the angiotensin-converting enzyme inhibitor enalapril or their combination have been incompletely studied in a large cohort of patients with heart failure not treated with beta-blockers. The objective of this study was to investigate the changes in neurohormones and LV volumes and ejection fraction in patients treated with enalapril, candesartan, or enalapril plus candesartan without concomitant beta-blocker therapy. METHODS: Three hundred and ninety-two patients from the RESOLVD pilot study not treated with a beta-blocker at baseline or at any time during the trial were analyzed. Norepinephrine, endothelin-1, big endothelin-1, angiotensin-II, aldosterone, N-terminal proANP, BNP, and radionuclide angiography were measured before and after 43 weeks of treatment with candesartan alone (n = 162), or enalapril alone (n = 45), or candesartan plus enalapril (n = 185). Endpoints were assessed at baseline and after 43 weeks of therapy. RESULTS: LV end-diastolic and end-systolic volumes increased significantly at 43 weeks in all groups except for patients treated with enalapril plus candesartan. BNP decreased at 43 weeks only in patients receiving dual angiotensin-II suppression (-6.1 +/- 37.8 pmol/l). Angiotensin-II levels were significantly increased in patients treated with candesartan (+23.6 +/- 47.1 pg/ml; p<0.05). CONCLUSION: We conclude that angiotensin-II modulation, with enalapril and candesartan, without concomitant utilization of beta-blocker lead to a decrease in BNP and an attenuation of the increase in LV end-diastolic and end-systolic volumes without a reversal of this process in the long term. PMID- 19299428 TI - Blood pressure responder rates versus goal rates: which metric matters? AB - Reducing blood pressure (BP) to guideline-recommended goals associated with reductions in cardiovascular risk is central to effective hypertension management. In addition to measuring BP reduction, clinical trials of antihypertensive agents should assess the percentage of patients responding to treatment. The Food and Drug Administration's defined rate of response required for drug approval is a reduction in diastolic BP (DBP) to <90 mmHg and/or a DBP reduction of > or = 10 mmHg. Consequently, some patients may be counted as responders even if they have not reached DBP <90 mmHg. An antihypertensive agent's effectiveness may be better assessed by the proportion of patients who achieve recommended BP goals. This article analyzes the frequency of response rates versus goal rates as endpoints in randomized trials since January 2001. Data showed that goal rates, especially combined systolic BP (SBP)/DBP goal rates, are consistently lower than response rates in studies evaluating both endpoints. Goal rates incorporating both SBP and DBP, or having a focus on SBP for individuals >50 years of age, provide the most clinically relevant information and are a more clinically relevant metric of an agent's ability to reduce BP than DBP alone. PMID- 19299429 TI - Coronary artery embolism from native mitral valve thrombus. PMID- 19299430 TI - C-reactive protein, infarct size, microvascular obstruction, and left-ventricular remodelling following acute myocardial infarction. AB - AIMS: This study assessed the relationship between inflammatory mediators and indices of infarct size and left-ventricular (LV) remodelling following successful primary percutaneous coronary intervention (PCI) in patients with first time ST elevation myocardial infarction (MI). METHODS AND RESULTS: Forty two patients admitted with an occluded single vessel were recruited consecutively. Cardiac magnetic resonance was used for serial assessment (2 days, 1 week, 2 months) of infarct size, microvascular obstruction (MO), and LV remodelling. Inflammatory mediators were analysed before and after PCI. Our major findings were: (1) Following PCI, there was a marked increase in plasma levels of C-reactive protein, closely correlated with an increase in interleukin-6 and terminal complement complex, reaching maximum 2 days after PCI; (2) C-reactive protein 2 days after PCI was significantly correlated with infarct size and parameters of LV remodelling 2 months after PCI; (3) Patients with persistent MO had significantly higher C-reactive protein levels 2 days following PCI. CONCLUSION: We suggest that the rapid increase in C-reactive protein levels in this model of successful revascularization of a single, totally occluded vessel reflects the degree of inflammation within the infarcted area. Our findings support a role for C-reactive protein-mediated complement activation as both a marker and mediator of myocardial damage following MI. Clinical study no.: NCT 00465868. PMID- 19299431 TI - From bone marrow to the arterial wall: the ongoing tale of endothelial progenitor cells. AB - Several physiological and pathophysiological stimuli or drugs modulate endothelial progenitor cell (EPC) mobilization. Moreover, levels of circulating EPCs predict cardiovascular risk and left ventricular remodelling after myocardial infarction. Nevertheless, our understanding in this field is complicated by lack of an unequivocal definition of EPCs, thus limiting their clinical applications. This review summarizes current knowledge and uncertainties on EPC characterization and mobilization in the attempt to define their role in the management of cardiovascular diseases. PMID- 19299432 TI - The targeting of cyclophilin D by RNAi as a novel cardioprotective therapy: evidence from two-photon imaging. AB - AIMS: An opening of the mitochondrial permeability transition pore (MPTP), which leads to the loss of mitochondrial membrane potential (DeltaPsi(m)), is the earliest event that commits cells to death, and this process is potentially a prime target for therapeutic intervention against myocardial ischaemia/reperfusion. We aimed to investigate the protective effects of RNA interference (RNAi)-mediated gene silencing of cyclophilin D (CypD), one of the putative components of the MPTP, against myocardial ischaemia/reperfusion using two-photon laser scanning microscopy. METHODS AND RESULTS: We created an adenovirus carrying short-interfering RNA (siRNA) that inactivates CypD. Transduction of CypD-siRNA in rat cardiomyocytes achieved a 61% reduction in CypD mRNA and a 63% reduction in protein levels as well as protection against oxidant induced DeltaPsi(m) loss and cytotoxicity. To further investigate the effects in vivo, we monitored the spatio-temporal changes of DeltaPsi(m) in perfused rat hearts subjected to ischaemia/reperfusion using two-photon imaging. Adult rats received direct intramyocardial injections of the adenovirus. Two to three days after injection, rat hearts were perfused by the Langendorff method and DeltaPsi(m) levels of individual cells were monitored. The progressive loss of DeltaPsi(m) during ischaemia/reperfusion was significantly suppressed in CypD siRNA-transduced cells compared with non-transduced cells. Furthermore, the protective effect of CypD-siRNA was dose-dependent. CONCLUSION: Therapeutic interventions designed to inactivate CypD may be a promising strategy for reducing cardiac injury against myocardial ischaemia/reperfusion. The two-photon imaging technique provides deeper insight into cardioprotective therapy that targets mitochondria. PMID- 19299433 TI - Activation of prostaglandin E2 EP1 receptor increases arteriolar tone and blood pressure in mice with type 2 diabetes. AB - AIMS: Type 2 diabetes mellitus is frequently associated with hypertension, but the underlying mechanisms are not completely understood. We tested the hypothesis that activation of type 1 prostaglandin E(2) (PGE(2)) receptor (EP1) increases skeletal muscle arteriolar tone and blood pressure in mice with type 2 diabetes. METHODS AND RESULTS: In 12-week-old, male db/db mice (with homozygote mutation in leptin receptor), systolic blood pressure was significantly elevated, compared with control heterozygotes. Isolated, pressurized gracilis muscle arterioles ( approximately 90 microm) of db/db mice exhibited an enhanced pressure- and angiotensin II (0.1-10 nM)-induced tone, which was reduced by the selective EP1 receptor antagonist, AH6809 (10 microM), to the level observed in arterioles of control mice. Exogenous application of PGE(2) (10 pM-100 nM) or the selective agonist of the EP1 receptor, 17-phenyl-trinor-PGE(2) (10 pM-100 nM), elicited arteriolar constrictions that were significantly enhanced in db/db mice (max: 31 +/- 4 and 29 +/- 5%), compared with controls (max: 20 +/- 2 and 14 +/- 3%, respectively). In the aorta of db/db mice, an increased protein expression of EP1, but not EP4, receptor was also detected by western immunoblotting. Moreover, we found that oral administration of the EP1 receptor antagonist, AH6809 (10 mg/kg/day, for 4 days), significantly reduced the systolic blood pressure in db/db, but not in control mice. CONCLUSION: Activation of EP1 receptors increases arteriolar tone, which could contribute to the development of hypertension in the db/db mice. PMID- 19299434 TI - Is neuromyelitis optica associated with human leukocyte antigen? AB - BACKGROUND: To establish whether or not multiple sclerosis (MS) and neuromyelitis optica (NMO) are different pathological entities, we wondered whether MS patients and NMO patients share the same pattern of human leukocyte antigen (HLA) predisposition. OBJECTIVE: To study a putative association between susceptibility to NMO and HLA class I or class II loci in Caucasians. METHODS: A total of 39 unrelated Caucasian patients with NMO and six patients at a high risk of converting to NMO were studied. DNA genotyping of HLA class I and class II loci was assessed and allelic frequencies were reported at a high-resolution level. A case-control study by comparing the allelic distribution in the NMO patients with that of a French Caucasian MS group and a French Caucasian healthy group was carried out. RESULTS: The frequencies of HLA-DQA1, DQB1, and HLA-DRB1 DR2 alleles in the NMO group were intermediate between the healthy control group and the MS group. The DPB1*0501 allele was not increased in the NMO group compared with the healthy control group. The distribution of HLA-DRB1 allele enabled to distinguish between NMO-IgG-positive patients and healthy controls (P = 0.01). NMO-IgG negative patients presented an HLA II pattern closer to that of the MS group (P = 0.01). CONCLUSION: In contrast to the reported results in Asian opticospinal MS, we found no association between the DPB1*0501 allele and NMO in our Caucasian patients. Moreover, we suggest that NMO-IgG-positive patients could represent a distinct NMO group in terms of their genetic susceptibility. PMID- 19299435 TI - Serum IgG repertoire in clinically isolated syndrome predicts multiple sclerosis. AB - OBJECTIVE: We previously showed that serum IgG repertoires distinguished multiple sclerosis (MS) patients from healthy subjects and from patients with other inflammatory neurological diseases (OIND). We questioned whether the serum IgG repertoire of patients presenting a clinically isolated syndrome (CIS) could predict MS. METHODS: The global IgG immune responses against brain antigens in sera from 50 CIS patients were evaluated by immunoblotting. The IgG reactivities were compared with those from MS sera (n = 82), healthy sera (n = 27), and sera from OIND (n = 42). A linear discriminant analysis (LDA) defined a score for each individual. RESULTS: About 78% of scores obtained from CIS patients were located in the "MS area." During the follow-up (3.5 +/- 1.3 years), 28 patients fulfilled the McDonald criteria for MS, 15 patients remained CIS, and 7 patients developed OIND. Among the patients with an LDA score in the "MS area," 61.5% converted to MS. DISCUSSION: Our results suggest that a pathological distortion of the self reactive IgG repertoire occurs early so that immunomodulating treatment should be started as early as possible; they also highlight the early involvement of B cells in the physiopathological process in MS. PMID- 19299436 TI - Neuromyelitis optica in Brazil: a study on clinical and prognostic factors. AB - OBJECTIVES: To describe the clinical characteristics of patients with relapsing neuromyelitis optica (NMO) from a tertiary care center in Brazil and compare the groups with normal and abnormal brain magnetic resonance imaging (MRI). METHODS: Retrospective review of 41 patients followed at the Neuroimmunology Clinic of the Federal University of Sao Paulo, Brazil, from 1994 to 2007. RESULTS: All patients had relapsing-remitting optic-spinal disease, long extending spinal cord lesions, and brain MRI not meeting Barkhof criteria for multiple sclerosis (MS), thus fulfilling the 1999 and 2006 Wingerchuck criteria for NMO. Mean follow-up time was 52 months; mean age of onset was 32.6 years. The mean relapse rate (RR) and progression index (PI) were 1.0 and 0.9, respectively. Twenty-four patients had brain lesions not compatible with MS on MRI, and there were no statistical differences on PI and RR between patients who had brain lesions and patients who did not. Incomplete recovery, but not the type of first relapse, correlated with a worse prognosis. Seventeen patients were tested for NMO-IgG (anti-aquaporin-4 antibody) with 41% positivity. CONCLUSIONS: In this series, we did not find a statistical difference of disease progression between patients with and without brain lesions, suggesting that the presence of brain abnormalities is not a marker of disease severity. PMID- 19299437 TI - Epidemiological characteristics of pregnancy, delivery, and birth outcome in women with multiple sclerosis in Argentina (EMEMAR study). AB - BACKGROUND: The influence of pregnancy on Multiple Sclerosis (MS) has been extensively studied but such influence on Latin American women with MS has not been characterized. Our objective was to describe the course of pregnancy and birth outcome in Argentinean MS patients and the evolution of MS during pregnancy and after delivery. METHOD: We used a retrospective design in eight MS centers in Argentina and administered a survey to women with definite MS (Mc Donald) with pregnancies during or after MS onset. We contacted 355 women of which 81 met inclusion criteria. We recorded 141 pregnancies. RESULTS: Involuntary abortion was observed in 16% of pregnancies (95% CI = 10-23). Thirty five women received immunomodulatory therapy (IMT) before 42 pregnancies. Twenty three (55%) out of 42 pregnancies were exposed to IMT. The mean time of IMT discontinuation before conception in 19 (45.2%) pregnancies without exposure, was 104 days (95% CI = 61.0-147.0). There were 103 deliveries: 79% full term. Birth defects were detected in 19% of pregnancies exposed to IMT (95% CI = 4-46) and in 2% of non exposed (95% CI = 0.3-8.0). The mean relapse rate was: pre-pregnancy year: 0.22 (95% CI = 0.12-0.32); pregnancy: 0.31 in 1st (95% CI = 0.10-0.52), 0.19 (95% CI = 0.03-0.36) in 2nd, and 0.04 in 3rd trimester (95% CI = -0.04-0.12); 1st trimester post delivery: 0.82 (95% CI = 0.42-1.22). CONCLUSION: We observed a higher rate of birth defects among infants exposed to immunomodulators in utero than those not exposed. The reduction in MS relapses during 2nd and 3rd trimester of pregnancy and its increase during postpartum is consistent with previous reports. PMID- 19299438 TI - Depressive symptoms and coping in newly diagnosed patients with multiple sclerosis. AB - BACKGROUND: Multiple sclerosis (MS) is a chronic disease with unclear etiology, unpredictable clinical course, and no cure. Patients' ability to cope with MS moderates the adaptation to the disease. OBJECTIVES: To compare coping in patients recently diagnosed with MS and healthy controls and to study the association between depressive symptoms and patients' coping styles. METHODS: A sample of 86 recently diagnosed patients with definite or probable MS and 93 healthy population controls completed questionnaires assessing coping styles and depressive symptoms. RESULTS: Compared with healthy controls, patients with MS used significantly less the problem focused strategies including planning, restraint coping, and seeking social support for instrumental reasons, and they used less the emotion-focused strategies seeking social support for emotional reasons, focusing on and venting of emotions, and positive reinterpretation and growth. The mean Beck Depressive symptoms Inventory scores were 10.8 and 4.7 in patients and controls, respectively. In stress situations connected to MS, depressive symptoms in these patients were related to the problem-focused strategies of restraint coping and planning, the emotion-focused strategy of focusing on and venting of emotions, and the avoidance strategies of behavioral- and mental disengagements, and denial. PMID- 19299439 TI - The clinical effect of neutralizing antibodies against interferon-beta is independent of the type of interferon-beta used for patients with relapsing remitting multiple sclerosis. AB - OBJECTIVE: To establish whether the clinical effect of neutralizing antibodies (NAbs) against interferon-beta (IFN beta) depends on the type of IFNbeta (1a or 1b) used for treatment of patients with relapsing-remitting multiple sclerosis (MS). INTRODUCTION: NAbs against IFN beta-1b appear faster and may be more evenly distributed on IgG subclasses, whereas NAbs against IFN beta-1a develop more slowly and may be devoid of IgG3. This might cause different clinical responses to NAbs. DESIGN/PATIENTS: All Danish MS-patients who had started first-time treatment with IFNbeta-1a 22 microg s.c tiw (Rebif22) or IFN beta-1b 250 microg s.c. qod (Betaferon) before January 1st 2003 were included. Relapses were recorded at bi-annual visit. METHODS: We measured NAbs every 12 months using a clinically validated cytopathic effect assay. A blood sample with a neutralizing capacity of 20% or more was considered as NAb-positive. We used a mixed logistic regression analysis in which NAb-status (three levels), IFN beta-preparation, and time since treatment started were included as explanatory variables, and relapse rate as response variable. RESULTS: In 1,309 patients, who were observed for 21,958 months, 32.3% were classified as NAb-positive. The odds-ratio (OR) for relapses in NAb-positive months compared with NAb-negative months was 1.25; P = 0.02. The risk of relapses was higher with Betaferon than with Rebif22 (OR 1.26; P < 0.01). The effect of NAb-level on relapses was independent of whether the patients were treated with Betaferon or Rebif22 (P = 0.89) and of time (P = 0.80). CONCLUSION: NAbs caused by IFNbeta-1a s.c. do not differ from NAbs caused by IFNbeta-1b in their detrimental clinical effect. PMID- 19299440 TI - Demographics of pediatric-onset multiple sclerosis in an MS center population from the Northeastern United States. AB - BACKGROUND: The prevalence of pediatric-onset multiple sclerosis (MS) in the United States is unknown. OBJECTIVE: In a large cohort of MS patients, we sought to identify the proportion with first symptom-onset below the age of 18 years, and to compare their demographic and disease characteristics to a typical adult onset MS population. METHODS: Patients seen at the Partners Multiple Sclerosis Center, Brigham and Women's Hospital, Boston, Massachusetts, with clinical histories and characteristics of first symptoms recorded in an electronic database, were included in this study. RESULTS: We found that 3.06% of patients with a recorded MS history experienced a first attack under the age of 18 years of age compared to 30.83% of patients who experienced first symptoms between the ages of 25-35 years. Gender proportions were similar in both groups, with the exception of a lower female preponderance in pre-pubertal-onset patients. There was a higher proportion of non-Caucasians in the younger cohort. Localization of first symptoms was similar in the two groups. CONCLUSION: About 3% of MS patients experience their first symptom prior to the age of 18 years. Standardized follow up is required after a first demyelinating attack in childhood, which may lead to earlier diagnosis and treatment of pediatric-onset MS. PMID- 19299441 TI - Brain activation in multiple sclerosis: a BOLD fMRI study of the effects of fatiguing hand exercise. AB - BACKGROUND: Multiple sclerosis (MS) patients experience fatigue as a chronic symptom that decreases quality of life. Commonly, fatigue in MS patients is manifested as decreased motor function during or after physical activity and is associated with changes in brain metabolism. OBJECTIVE: To determine brain activation patterns in MS patients and healthy controls during a simple motor task before and after fatiguing hand-grip exercise. METHODS: Functional magnetic resonance imaging (fMRI) scans were conducted on 10 MS patients and 13 healthy controls during 4-finger flexion and extension in rested and fatigued states. RESULTS: Before the fatigue protocol, MS patients had greater activation in the contralateral primary motor cortex, insula, and cingulate gyrus than controls. Following fatiguing exercise, controls showed increased activation of precentral gyrus and insula while patients did not show any activation increases and actually decreased activity to the insula. CONCLUSION: Results indicate that before fatiguing exercise, MS patients marshaled more brain activation compared to controls, which may represent functionally adaptive changes in response to demyelination. This increased activation may suggest that patients require more effort to perform even simple motor tasks, possibly because peripheral or central signals for fatigue are chronically enhanced. When fatigued further by muscle contraction, brain activation cannot be further increased. PMID- 19299442 TI - The DREAM protein is associated with thyroid enlargement and nodular development. AB - G protein-coupled receptors (GPCRs) are involved in the pathophysiology of a wide range of diseases and constitute an attractive therapeutic target. In the thyroid gland, TSH receptor (TSHR), a member of the GPCR family, is a major regulator of thyroid differentiation and function. Alterations in TSHR activity are often involved in the development of pathologies such as thyroid cancer and thyroid enlargement (goiter). Here we show that DREAM (downstream regulatory element antagonist modulator) modulates TSHR activity through a direct protein-protein interaction that promotes coupling between the receptor and Galphas. In transgenic mice, DREAM overexpression provokes a marked enlargement of the thyroid gland. Increased levels of DREAM protein were observed in human multinodular goiters, suggesting a novel etiopathogenic mechanism in nodular development in humans. Taken together, these findings identify a mechanism for the control of TSHR activity and provide a new approach for the study and treatment of thyroid pathologies associated with impaired TSHR function. PMID- 19299443 TI - Mutational analysis of progesterone receptor functional domains in stable cell lines delineates sets of genes regulated by different mechanisms. AB - Steroid hormone receptors act directly in the nucleus on the chromatin organization and transcriptional activity of several promoters. Furthermore, they have an indirect effect on cytoplasmic signal transduction pathways, including MAPK, impacting ultimately on gene expression. We are interested in distinguishing between the two modes of action of progesterone receptor (PR) on the control of gene expression and cell proliferation. For this, we have stably expressed, in PR-negative breast cancer cells, tagged forms of the PR isoform B mutated at regions involved either in DNA binding (DNA-binding domain) or in its ability to interact with the estrogen receptor and to activate the c Src/MAPK/Erk/Msk cascade (estrogen receptor-interacting domain). Both mutants impair PR-mediated activation of a well-understood model promoter in response to progestin, as well as hormone-induced cell proliferation. Additional mutants affecting transactivation activity of PR (activation function 2) or a zinc-finger implicated in dimerization (D-box) have also been tested. Microarrays and gene expression experiments on these cell lines define the subsets of hormone responsive genes regulated by different modes of action of PR isoform B, as well as genes in which the nuclear and nongenomic pathways cooperate. Correlation between CCND1 expression in the different cell lines and their ability to support cell proliferation confirms CCND1 as a key controller gene. PMID- 19299444 TI - Ghrelin induces abdominal obesity via GHS-R-dependent lipid retention. AB - Circulating ghrelin elevates abdominal adiposity by a mechanism independent of its central orexigenic activity. In this study we tested the hypothesis that peripheral ghrelin induces a depot-specific increase in white adipose tissue (WAT) mass in vivo by GH secretagogue receptor (GHS-R(1a))-mediated lipolysis. Chronic iv infusion of acylated ghrelin increased retroperitoneal and inguinal WAT volume in rats without elevating superficial sc fat, food intake, or circulating lipids and glucose. Increased retroperitoneal WAT mass resulted from adipocyte enlargement probably due to reduced lipid export (ATP-binding cassette transporter G1 mRNA expression and circulating free fatty acids were halved by ghrelin infusion). In contrast, ghrelin treatment did not up-regulate biomarkers of adipogenesis (peroxisome proliferator-activated receptor-gamma2 or CCAAT/enhancer binding protein-alpha) or substrate uptake (glucose transporter 4, lipoprotein lipase, or CD36) and although ghrelin elevated sterol-regulatory element-binding protein 1c expression, WAT-specific mediators of lipogenesis (liver X receptor-alpha and fatty acid synthase) were unchanged. Adiposity was unaffected by infusion of unacylated ghrelin, and the effects of acylated ghrelin were abolished by transcriptional blockade of GHS-R(1a), but GHS-R(1a) mRNA expression was similar in responsive and unresponsive WAT. Microarray analysis suggested that depot-specific sensitivity to ghrelin may arise from differential fine tuning of signal transduction and/or lipid-handling mechanisms. Acylated ghrelin also induced hepatic steatosis, increasing lipid droplet number and triacylglycerol content by a GHS-R(1a)-dependent mechanism. Our data imply that, during periods of energy insufficiency, exposure to acylated ghrelin may limit energy utilization in specific WAT depots by GHS-R(1a)-dependent lipid retention. PMID- 19299445 TI - Estrogen receptor alpha (ERalpha) mediates stimulatory effects of estrogen on aromatase (CYP19) gene expression in human placenta. AB - A 246-bp region upstream of placenta-specific exon I.1 of the human aromatase (hCYP19) gene mediates placenta-specific, developmental, and O(2) regulation of expression. In this study, trophoblast differentiation and associated induction of CYP19 expression were prevented when cytotrophoblasts were cultured in phenol red-free medium containing charcoal-stripped serum or with the estrogen receptor (ER) antagonist, ICI 182,780, suggesting a stimulatory role of estrogen/ER. ERalpha protein was expressed in human trophoblasts and increased during syncytiotrophoblast differentiation, whereas ERbeta was undetectable. Mutational analysis revealed that an estrogen response element-like sequence (ERE-LS) at 208 bp is required for inductive effects of estradiol/ERalpha on hCYP19I.1 promoter activity in transfected COS-7 cells. Increased binding of syncytiotrophoblast compared with cytotrophoblast nuclear proteins to the ERE-LS was observed in vitro; however, ERalpha antibodies failed to supershift the complex and in vitro-transcribed/translated ERalpha did not bind. Nonetheless, chromatin immunoprecipitation assays in cultured trophoblasts revealed recruitment of endogenous ERalpha to the -255- to -155-bp region containing the ERE-LS before induction of hCYP19 expression; this was inhibited by ICI 182,780. Chromatin immunoprecipitation also revealed increased acetylated histone H3(K9/14) and decreased methylated histone H3(K9) associated with this region during trophoblast differentiation. These modifications were prevented when trophoblasts were incubated with ICI 182,780, suggesting that ERalpha recruitment to the -255- to -155-bp region promotes histone modifications leading to increased hCYP19 transcription. Thus, during trophoblast differentiation, estrogen/ERalpha exerts a positive feedback role, which promotes permissive histone modifications that are associated with induction of hCYP19 gene transcription. PMID- 19299446 TI - Glutaredoxin-1 mediates NADPH-dependent stimulation of calcium-dependent insulin secretion. AB - Nicotinamide adenine dinucleotide phosphate (NADPH) enhances Ca(2+)-induced exocytosis in pancreatic beta-cells, an effect suggested to involve the cytosolic redox protein glutaredoxin-1 (GRX-1). We here detail the role of GRX-1 in NADPH stimulated beta-cell exocytosis and glucose-stimulated insulin secretion. Silencing of GRX-1 by RNA interference reduced glucose-stimulated insulin secretion in both clonal INS-1 832/13 cells and primary rat islets. GRX-1 silencing did not affect cell viability or the intracellular redox environment, suggesting that GRX-1 regulates the exocytotic machinery by a local action. By contrast, knockdown of the related protein thioredoxin-1 (TRX-1) was ineffective. Confocal immunocytochemistry revealed that GRX-1 locates to the cell periphery, whereas TRX-1 expression is uniform. These data suggest that the distinct subcellular localizations of TRX-1 and GRX-1 result in differences in substrate specificities and actions on insulin secretion. Single-cell exocytosis was likewise suppressed by GRX-1 knockdown in both rat beta-cells and clonal 832/13 cells, whereas after overexpression exocytosis increased by approximately 40%. Intracellular addition of NADPH (0.1 mm) stimulated Ca(2+)-evoked exocytosis in both cell types. Interestingly, the stimulatory action of NADPH on the exocytotic machinery coincided with an approximately 30% inhibition in whole-cell Ca(2+) currents. After GRX-1 silencing, NADPH failed to amplify insulin release but still inhibited Ca(2+) currents in 832/13 cells. In conclusion, NADPH stimulates the exocytotic machinery in pancreatic beta-cells. This effect is mediated by the NADPH acceptor protein GRX-1 by a local redox reaction that accelerates beta-cell exocytosis and, in turn, insulin secretion. PMID- 19299448 TI - Hypermethylation of homeobox A10 by in utero diethylstilbestrol exposure: an epigenetic mechanism for altered developmental programming. AB - Diethylstilbestrol (DES) is a nonsteroidal estrogen that induces developmental anomalies of the female reproductive tract. The homeobox gene HOXA10 controls uterine organogenesis, and its expression is altered after in utero DES exposure. We hypothesized that an epigenetic mechanism underlies DES-mediated alterations in HOXA10 expression. We analyzed the expression pattern and methylation profile of HOXA10 after DES exposure. Expression of HOXA10 is increased in human endometrial cells after DES exposure, whereas Hoxa10 expression is repressed and shifted caudally from its normal location in mice exposed in utero. Cytosine guanine dinucleotide methylation frequency in the Hoxa10 intron was higher in DES exposed offspring compared with controls (P = 0.017). The methylation level of Hoxa10 was also higher in the caudal portion of the uterus after DES exposure at the promoter and intron (P < 0.01). These changes were accompanied by increased expression of DNA methyltransferases 1 and 3b. No changes in methylation were observed after in vitro or adult DES exposure. DES has a dual mechanism of action as an endocrine disruptor; DES functions as a classical estrogen and directly stimulates HOXA10 expression with short-term exposure, however, in utero exposure results in hypermethylation of the HOXA10 gene and long-term altered HOXA10 expression. We identify hypermethylation as a novel mechanism of DES-induced altered developmental programming. PMID- 19299447 TI - Effects of soy protein and isoflavones on circulating hormone concentrations in pre- and post-menopausal women: a systematic review and meta-analysis. AB - BACKGROUND Hormonal effects of soy and isoflavones have been investigated in numerous trials with equivocal findings. We aimed to systematically assess the effects of soy and isoflavones on circulating estrogen and other hormones in pre- and post-menopausal women. METHODS The Cochrane Library, MEDLINE and EMBASE (plus reviews and experts) were searched to December 2007. Inclusion of randomized or residential crossover trials of soy or isoflavones for 4 or more weeks on estrogens, SHBG, FSH, LH, progesterone and thyroid hormones in women was assessed independently in duplicate. Six percent of papers assessed were included. Data concerning participants, interventions, outcomes, potential effect modifiers and trial quality characteristics were extracted independently in duplicate. RESULTS Forty-seven studies (11 of pre-, 35 of post- and 1 of perimenopausal women) were included. In premenopausal women, meta-analysis suggested that soy or isoflavone consumption did not affect primary outcomes estradiol, estrone or SHBG concentrations, but significantly reduced secondary outcomes FSH and LH [by approximately 20% using standardized mean difference (SMD), P = 0.01 and 0.05, respectively]. Menstrual cycle length was increased by 1.05 days (95% CI 0.13, 1.97, 10 studies). In post-menopausal women, there were no statistically significant effects on estradiol, estrone, SHBG, FSH or LH, although there was a small statistically non-significant increase in total estradiol with soy or isoflavones ( approximately 14%, SMD, P = 0.07, 21 studies). CONCLUSIONS Isoflavone-rich soy products decrease FSH and LH in premenopausal women and may increase estradiol in post-menopausal women. The clinical implications of these modest hormonal changes remain to be determined. PMID- 19299449 TI - The corpuscles of Stannius, calcium-sensing receptor, and stanniocalcin: responses to calcimimetics and physiological challenges. AB - This study has examined whether the calcium-sensing receptor (CaSR) plays a role in control of stanniocalcin-1 (STC-1), the dominant calcium regulatory hormone of fish, comparable with that demonstrated for CaSR in the mediation of ionized calcium regulation of PTH secretion in mammals. In a previous study, we have cloned flounder STC-1 from the corpuscles of Stannius (CS). Here, we report the cloning and characterization of the CS CaSR, and the in vivo responses of this system to altered salinity, EGTA induced hypocalcemia, and calcimimetic administration. Quantitative PCR analysis demonstrated, for the first time, that the CS are major sites of CaSR expression in flounder. Immunoblot analysis of CS proteins with CaSR-specific antibodies revealed a broad band of approximately 215 300 kDa under nonreducing conditions, and bands of approximately 215-300 kDa and approximately 120-150 kDa under reducing conditions. There were no differences in CS CaSR mRNA expression or plasma STC-1 levels between seawater and freshwater (FW)-adapted fish, although CS STC-1 mRNA expression was lower in FW animals. Immunoblots showed that glycosylated monomeric forms of the CaSR migrated at a lower molecular mass in CS samples from FW animals. The ip administration of EGTA rapidly induced hypocalcemia, and a concomitant lowering of plasma STC-1. Calcimimetic administration (1 mg/kg R-568) rapidly increased plasma STC-1 levels, and reduced plasma concentrations of calcium, phosphate, and magnesium when compared with S-568-treated controls. Together, these findings support an evolutionary conserved role for the CaSR in the endocrine regulation of calcium before the appearance of parathyroid glands in tetrapods. PMID- 19299450 TI - Hormonal modulation of amino acid neurotransmitter metabolism in the arcuate nucleus of the adult female rat: a novel action of estradiol. AB - Morphological plasticity in response to estradiol is a hallmark of astrocytes in the arcuate nucleus. The functional consequences of these morphological changes have remained relatively unexplored. Here we report that in the arcuate nucleus estradiol significantly increased the protein levels of the two enzymes in the glutamate-glutamine cycle, glutamine synthetase and glutaminase. We further demonstrate that these estradiol-mediated changes in the enzyme protein levels may underlie functional changes in neurotransmitter availability as: 1) total glutamate concentration in the arcuate nucleus was significantly increased and 2) microdialysis revealed a significant increase in extracellular glutamate levels after a synaptic challenge in the presence of estradiol. These data implicate the glutamate-glutamine cycle in the generation and/or maintenance of glutamate and suggest that the difference in extracellular glutamate between estradiol- and oil treated animals may be related to an increased efficiency of the cycle enzymes. In vivo enzyme activity assays revealed that the estradiol mediated increase in glutamate-glutamine cycle enzymes in the arcuate nucleus led to an increase in gamma-aminobutyric acid and is likely not related to the increase in extracellular glutamate. Thus, we have observed two-independent effects of estradiol on amino acid neurotransmission in the arcuate nucleus. These data suggest a possible functional consequence of the well-established changes in glial morphology that occur in the arcuate nucleus in the presence of estradiol and suggest the importance of neuronal-glial cooperation in the regulation of hypothalamic functions such as food intake and body weight. PMID- 19299451 TI - Curcumin eliminates leptin's effects on hepatic stellate cell activation via interrupting leptin signaling. AB - Nonalcoholic steatohepatitis (NASH) is commonly found in patients with obesity and is often accompanied with abnormally elevated levels of plasma leptin, i.e. hyperleptinemia. A relatively high population of NASH patients develops hepatic fibrosis, even cirrhosis. Hepatic stellate cells (HSCs) are the major effector cells during liver fibrogenesis and could be activated by leptin. The antioxidant curcumin, a phytochemical from turmeric, has been shown to suppress HSC activation in vitro and in vivo. This project is to evaluate the effect of curcumin on leptin-induced HSC activation and to elucidate the underlying mechanisms. We hypothesize that curcumin abrogates the stimulatory effect of leptin on HSC activation by interrupting leptin signaling and attenuating leptin induced oxidative stress. Curcumin eliminates the stimulatory effects of leptin on regulating expression of genes closely relevant to HSC activation. Curcumin interrupts leptin signaling by reducing phosphorylation levels of leptin receptor (Ob-R) and its downstream intermediators. In addition, curcumin suppresses gene expression of Ob-R in HSCs, which requires the activation of endogenous peroxisome proliferator-activated receptor-gamma and de novo synthesis of glutathione. In conclusion, our results demonstrate that curcumin abrogates the stimulatory effect of leptin on HSC activation in vitro by reducing the phosphorylation level of Ob-R, stimulating peroxisome proliferator-activated receptor-gamma activity, and attenuating oxidative stress, leading to the suppression of Ob-R gene expression and interruption of leptin signaling. These results provide novel insights into therapeutic mechanisms of curcumin in inhibiting HSC activation and intervening liver fibrogenesis associated with hyperleptinemia in NASH patients. PMID- 19299452 TI - Unique profile of chicken adiponectin, a predominantly heavy molecular weight multimer, and relationship to visceral adiposity. AB - Adiponectin, a 30-kDa adipokine hormone, circulates as heavy, medium, and light molecular weight isoforms in mammals. Plasma heavy molecular weight (HMW) adiponectin isoform levels are inversely correlated with the incidence of type 2 diabetes in humans. The objectives of the present study were to characterize adiponectin protein and quantify plasma adiponectin levels in chickens, which are naturally hyperglycemic relative to mammals. Using gel filtration column chromatography and Western blot analysis under nonreducing and non-heat denaturing native conditions, adiponectin in chicken plasma, and adipose tissue is predominantly a multimeric HMW isoform that is larger than 669 kDa mass. Under reducing conditions and heating to 70-100 C, however, a majority of the multimeric adiponectin in chicken plasma and adipose tissue was reduced to oligomeric and/or monomeric forms. Immunoprecipitation and elution under neutral pH preserved the HMW adiponectin multimer, whereas brief exposure to acidic pH led to dissociation of HMW multimer into multiple oligomers. Mass spectrometric analysis of chicken adiponectin revealed the presence of hydroxyproline and differential glycosylation of hydroxylysine residues in the collagenous domain. An enzyme immunoassay was developed and validated for quantifying plasma adiponectin in chickens. Plasma adiponectin levels were found to be significantly lower in 8- compared with 4-wk-old male chickens and inversely related to abdominal fat pad mass. Collectively, our results provide novel evidence that adiponectin in chicken plasma and tissues is predominantly a HMW multimer, suggesting the presence of unique multimerization and stabilization mechanisms in the chicken that favors preponderance of HMW adiponectin over other oligomers. PMID- 19299453 TI - Interleukin-6: an autocrine regulator of the mouse cumulus cell-oocyte complex expansion process. AB - Ovulation has long been regarded as a process resembling an inflammatory response. Recent studies indicate that genes associated with innate immune responses were also expressed during the ovulation process. Because the innate immune genes are induced in cumulus cell oocyte complexes (COCs) later than the inflammation-associated genes, we hypothesize that COC expansion is dependent on specific sequential changes in cumulus cells. Because IL-6 is a potent mediator of immune responses, we sought to determine what factors regulate the induction of Il6 mRNA in COCs and what impact IL-6 alone would have on COC expansion. We found that the levels of Il6 mRNA increased dramatically during COC expansion, both in vivo and in vitro. Moreover, IL-6, together with its soluble receptor (IL 6SR), could bypass the need for either amphiregulin and/or prostaglandin E2 to induce the expansion of COCs. This ability of IL-6/IL-6SR to induce COC expansion was blocked by the inhibitors to p38MAPK, MAPK kinase 1/2, and Janus kinase. More importantly, when COCs were in vitro maturated in the presence of IL-6, they had a significantly higher embryo transfer rate than the ones without IL-6 and comparable with in vivo matured oocytes. IL-6/IL-6SR activated multiple signaling pathways (Janus kinase/signal transducer and activator of transcription, ERK1/2, p38MAPK, and AKT) and progressively induced genes known to impact COC expansion, genes related to inflammation and immune responses, and some transcription factors. Collectively, these data indicate that IL-6 alone can act as a potent autocrine regulator of ovarian cumulus cell function, COC expansion, and oocyte competence. PMID- 19299454 TI - Up-regulation of cyclooxygenase-2 expression and prostaglandin E2 production in human endometriotic cells by macrophage migration inhibitory factor: involvement of novel kinase signaling pathways. AB - Cyclooxygenase (COX) is the rate-limiting enzyme in the metabolic conversion of arachidonic acid to prostaglandins (PGs), including prostaglandin E(2) (PGE(2)), a major mediator of inflammation and angiogenesis. Herein, we report that macrophage migration inhibitory factor (MIF), a potent proinflammatory and growth promoting factor found at elevated concentrations in the peritoneal fluid of women with endometriosis and active endometriosis lesions, acts directly on ectopic endometrial cells to stimulate the synthesis of COX-2, the inducible form of COX, and the release of PGE(2). MIF treatment strongly activated p38 and ERK MAPK, and specific inhibitors of both pathways completely blocked basal and MIF induced PGE(2) synthesis. Whereas p38 inhibitors negatively affected the stimulated synthesis of COX-2 and that of PGE(2), ERK inhibitors only decreased the production of PGE(2). These findings show for the first time a direct role for MIF in the up-regulation of COX-2 synthesis and PGE(2) secretion in ectopic endometrial cells. They further indicate that whereas p38 and ERK MAPK signaling pathways both play a significant role in the regulation of basal and MIF-induced synthesis of PGE(2) by ectopic endometrial cells, only p38 kinase is involved in the regulation of COX-2 expression in these cells. This suggests that MIF acts at more than one level to stimulate the synthesis of PGE(2) and triggers the coordinate activation of multiple enzymes in the biosynthesis pathway. Our data provide evidence for a novel mechanism by which MIF can induce a proinflammatory phenotype in ectopic endometrial cells, and favor the establishment of endometriosis and its related clinical symptoms. PMID- 19299455 TI - Perilipin is present in islets of Langerhans and protects against lipotoxicity when overexpressed in the beta-cell line INS-1. AB - Lipids have been shown to play a dual role in pancreatic beta-cells: a lipid derived signal appears to be necessary for glucose-stimulated insulin secretion, whereas lipid accumulation causes impaired insulin secretion and apoptosis. The ability of the protein perilipin to regulate lipolysis prompted an investigation of the presence of perilipin in the islets of Langerhans. In this study evidence is presented for perilipin expression in rat, mouse, and human islets of Langerhans as well as the rat clonal beta-cell line INS-1. In rat and mouse islets, perilipin was verified to be present in beta-cells. To examine whether the development of lipotoxicity could be prevented by manipulating the conditions for lipid storage in the beta-cell, INS-1 cells with adenoviral-mediated overexpression of perilipin were exposed to lipotoxic conditions for 72 h. In cells exposed to palmitate, perilipin overexpression caused increased accumulation of triacylglycerols and decreased lipolysis compared with control cells. Whereas glucose-stimulated insulin secretion was retained after palmitate exposure in cells overexpressing perilipin, it was completely abolished in control beta-cells. Thus, overexpression of perilipin appears to confer protection against the development of beta-cell dysfunction after prolonged exposure to palmitate by promoting lipid storage and limiting lipolysis. PMID- 19299456 TI - The acid-labile subunit is required for full effects of exogenous growth hormone on growth and carbohydrate metabolism. AB - Normal postnatal growth is dependent in part on overlapping actions of GH and IGF I. These actions reflect GH stimulation of IGF-I production in liver and extrahepatic tissues, representing respectively the endocrine and autocrine/paracrine arms of the IGF system. Recent experiments in genetically modified mice show that each source of IGF-I can compensate for absence of the other but do not resolve their relative role in postnatal growth. In an effort to address this issue, we studied the GH responsiveness of mice harboring a null mutation of the acid-labile subunit (ALS). Null ALS mice have a substantial reduction in endocrine IGF-I but, unlike other models of plasma IGF-I deficiency, have no obvious additional endocrine defects. Wild type and null ALS mice of both sexes received daily sc injections of saline or recombinant bovine GH between d 35 and 63 of postnatal age. The GH-stimulated body weight gain of null ALS mice was reduced by more than 30% relative to wild type mice, irrespective of sex. Reductions in GH responsiveness were also seen for kidney and linear growth. Absence of ALS eliminated the ability of GH to increase plasma IGF-I despite intact GH-dependent stimulation of IGF-I expression in liver, adipose tissue, and skeletal muscle. GH treatment was also less efficient in antagonizing insulin action in null ALS mice. Overall, these results suggest that the GH effects mediated by endocrine IGF-I depends on ALS, and accordingly null ALS mice are less responsive to exogenous GH therapy. PMID- 19299457 TI - A monoclonal antibody with thyrotropin (TSH) receptor inverse agonist and TSH antagonist activities binds to the receptor hinge region as well as to the leucine-rich domain. AB - Monoclonal antibody CS-17 is a TSH receptor (TSHR) inverse agonist (suppresses constitutive activity) and a TSH antagonist. Elucidation of the CS-17 epitope will provide insight into TSHR structure and function. Present information on its epitope conflicts with recent data regarding another TSHR inverse agonist antibody. To characterize further the CS-17 epitope, we exploited the observation that CS-17 does not recognize a chimeric receptor with TSHR hinge region residues 261-289 replaced with homologous rat LH receptor residues (13 mismatches). We generated individual and double TSHR mutations corresponding to these mismatches. On flow cytometry, only T273L/R274V reduced CS-17 recognition. No mutation affected TSH-stimulated cAMP generation. Because the immunogen for CS-17 generation was highly glycosylated, we also investigated whether the glycan moiety at N198, topologically adjacent to Y195 (a previously identified epitopic component), could contribute to the CS-17 epitope. Elimination of this N-linked glycan (mutations of N198 and T200) abrogated CS-17 binding without altering TSH responsiveness. However, studies with tunicamycin suggested that these mutations affected CS-17 binding by altering the polypeptide backbone rather than eliminating the glycan moiety. TSHR residues N198 and T200, like Y195, are on the convex facet of the leucine-rich domain. In summary, the present data indicate that the discontinuous epitope of CS-17, a TSHR inverse agonist and TSH antagonist, includes a component in the hinge region as well as the convex surface of the TSHR leucine-rich domain. These findings expand our present concept of glycoprotein hormone binding and function. PMID- 19299458 TI - Aberrant histone modifications at the thyrotropin-releasing hormone gene in resistance to thyroid hormone: analysis of F455S mutant thyroid hormone receptor. AB - We reported a novel mutation of thyroid hormone receptor (TR)-beta, F455S, in a patient with pituitary resistance to thyroid hormone (RTH), who showed impaired release of nuclear receptor corepressor and abnormal histone deacetylation. In the present study, we further analyzed the histone modifications and the dynamics of TR and RNA polymerase II on the TRH gene. The lysine residues 9 (H3K9) and 14 (K14) of the histone H3 were acetylated in the absence of thyroid hormone (TH), and addition of TH caused a temporary deacetylation of both residues. Although H3K4 was di- and trimethylated in the absence of T(3), no methylation of H3K9 or K27 was detected. Long-term incubation with T(3) decreased the level of trimethylated H3K4, the amount of TR, and the level of phosphorylated RNA polymerase II but not dimethylated H3K4. Treatment with an inhibitor for H3K4 methyltransferase, 5'-deoxy-5'-methylthioadenosine, decreased basal promoter activity but did not affect the repression by TH. Conversely, overexpression of MLL, an H3K4-specific methyltransferase, caused an increase in basal activity. In the presence of F455S, methylation of H3K4 and the dynamics of TR were intact, but both H3K9 and H3K14 were hyperacetylated, and T(3)-induced deacetylation was impaired, resulting in a high transcriptional level. These findings demonstrated that 1) negative regulation of the TRH gene by TH involves both the acetylation and methylation of specific residues of histone tails and changing the amount of TR, and 2) the major impairment to histone modifications in F455S was hyperacetylation of the specific histone tails. PMID- 19299459 TI - Postnatal development of an estradiol-kisspeptin positive feedback mechanism implicated in puberty onset. AB - The regulation of GnRH neurons by kisspeptin is critical for normal puberty onset in mammals. In the rodent the kisspeptin neurons innervating GnRH neurons are thought to reside in the rostral periventricular area of the third ventricle (RP3V). Using kisspeptin immunocytochemistry we show that kisspeptin peptide expression in the RP3V of female mice begins around postnatal d 15 (P15) and rapidly increases to achieve adult-like levels by P30, the time of puberty onset. Ovariectomy of female pups at P15 resulted in a 70-90% reduction (P < 0.01) in kisspeptin peptide expression within the RP3V of P30 or P60 mice. Replacement of 17-beta-estradiol (E2) in P15-ovariectomized mice from P15-30 or P22-30 resulted in a complete restoration of kisspeptin peptide expression in the RP3V (P < 0.01). Kisspeptin-immunoreactive fibers throughout the hypothalamus, including the arcuate nucleus, followed the same pattern of estrogen-dependent expression. To test the absolute necessity of estrogen for kisspeptin expression in the RP3V, aromatase knockout mice were examined. Kisspeptin-immunoreactive cells were detected in the arcuate nucleus, but there was a complete absence of kisspeptin peptide in RP3V neurons of aromatase knockout adult females. These results demonstrate that E2 is essential for the prepubertal development of kisspeptin peptide within RP3V neurons and suggest that an E2-kisspeptin positive feedback mechanism exists before puberty. This implies that RP3V kisspeptin neurons are E2 dependent amplifiers of GnRH neuron activity in the prepubertal period. PMID- 19299460 TI - Three-dimensional cellular architecture of the flagellar pocket and associated cytoskeleton in trypanosomes revealed by electron microscope tomography. AB - This study uses electron tomography linked to a variety of other EM methods to provide an integrated view of the flagellar pocket and basal body area of the African trypanosome procyclic trypomastigote. We reveal the pocket as an asymmetric membranous 'balloon' with two boundary structures. One of these - the collar - defines the flagellum exit point. The other defines the entry point of the flagellum into the pocket and consists of both an internal transitional fibre array and an external membrane collarette. A novel set of nine radial fibres is described in the basal body proximal zone. The pocket asymmetry is invariably correlated with the position of the probasal body and Golgi. The neck region, just distal to the flagellum exit site, is a specialised area of membrane associated with the start of the flagellum attachment zone and signifies the point where a special set of four microtubules, nucleated close to the basal bodies, joins the subpellicular array. The neck region is also associated with the single Golgi apparatus of the cell. The flagellar exit point interrupts the subpellicular microtubule array with discrete endings of microtubules at the posterior side. Overall, our studies reveal a highly organised, yet dynamic, area of cytoplasm and will be informative in understanding its function. PMID- 19299461 TI - Nucleocytoplasmic shuttling of soluble tubulin in mammalian cells. AB - We have investigated the subcellular distribution and dynamics of soluble tubulin in unperturbed and transfected HeLa cells. Under normal culture conditions, endogenous alpha/beta tubulin is confined to the cytoplasm. However, when the soluble pool of subunits is elevated by combined cold-nocodazole treatment and when constitutive nuclear export is inhibited by leptomycin B, tubulin accumulates in the cell nucleus. Transfection assays and FRAP experiments reveal that GFP-tagged beta-tubulin shuttles between the cytoplasm and the cell nucleus. Nuclear import seems to occur by passive diffusion, whereas exit from the nucleus appears to rely on nuclear export signals (NESs). Several such motifs can be identified by sequence criteria along the beta-tubulin molecule and mutations in one of these (NES-1) cause a significant accumulation in the nuclear compartment. Under these conditions, the cells are arrested in the G0-G1 phase and eventually die, suggesting that soluble tubulin interferes with important nuclear functions. Consistent with this interpretation, soluble tubulin exhibits stoichiometric binding to recombinant, normally modified and hyper-phosphorylated/acetylated histone H3. Tubulin-bound H3 no longer interacts with heterochromatin protein 1 and lamin B receptor, which are known to form a ternary complex under in vitro conditions. Based on these observations, we suggest that nuclear accumulation of soluble tubulin is part of an intrinsic defense mechanism, which tends to limit cell proliferation under pathological conditions. This readily explains why nuclear tubulin has been detected so far only in cancer or in transformed cells, and why accumulation of this protein in the nucleus increases after treatment with chemotherapeutic agents. PMID- 19299462 TI - Low-density lipoprotein receptor-related protein 1 is an essential receptor for myelin phagocytosis. AB - Multiple sclerosis (MS) is an autoimmune disease in which myelin is progressively degraded. Because degraded myelin may both initiate and accelerate disease progression, clearing degraded myelin from extracellular spaces may be critical. In this study, we prepared myelin vesicles (MV) from rat brains as a model of degraded myelin. Murine embryonic fibroblasts (MEFs) rapidly internalized MVs, which accumulated in lysosomes only when these cells expressed low-density lipoprotein receptor-related protein (LRP1). Receptor-associated protein (RAP), which binds LRP1 and inhibits interaction with other ligands, blocked MV uptake by LRP1-expressing MEFs. As a complementary approach, we prepared primary cultures of rat astrocytes, microglia and oligodendrocytes. All three cell types expressed LRP1 and mediated MV uptake, which was inhibited by RAP. LRP1 gene silencing in oligodendrocytes also blocked MV uptake. Myelin basic protein (MBP), which was expressed as a recombinant protein, bound directly to LRP1. MBP specific antibody inhibited MV uptake by oligodendrocytes. In experimental autoimmune encephalomyelitis in mice, LRP1 protein expression was substantially increased in the cerebellum and spinal cord. LRP1 colocalized with multiple CNS cell types. These studies establish LRP1 as a major receptor for phagocytosis of degraded myelin, which may function alone or in concert with co-receptors previously implicated in myelin phagocytosis. PMID- 19299463 TI - The nuclear pore component Nup358 promotes transportin-dependent nuclear import. AB - Nup358 (also known as RanBP2), a component of the cytoplasmic filaments of the nuclear pore complex, has been implicated in various nucleocytoplasmic transport pathways. Here, we identify Nup358 as an important factor for transportin mediated nuclear import. Depletion of Nup358 resulted in a strong inhibition of nuclear import of the human immunodeficiency virus type 1 (HIV-1) Rev protein. HIV-1 Rev is an RNA-binding protein that is required for CRM1 (also known as exportin 1)-dependent nuclear export of unspliced or partially spliced viral RNA. We show that transportin is the major nuclear import receptor for HIV-1 Rev in HeLa cells. Overexpression of transportin strongly promoted nuclear import of HIV 1 Rev in Nup358-depleted cells, indicating that the import receptor becomes rate limiting under these conditions. Importantly, the import rate of other transportin-dependent proteins was also significantly reduced in Nup358-depleted cells. Our data therefore suggest a general role for Nup358 in transportin mediated nuclear import. PMID- 19299464 TI - A Golgi-associated protein 4.1B variant is required for assimilation of proteins in the membrane. AB - The archetypal membrane skeleton is that of the erythrocyte, consisting predominantly of spectrin, actin, ankyrin R and protein 4.1R. The presence in the Golgi of a membrane skeleton with a similar structure has been inferred, based on the identification of Golgi-associated spectrin and ankyrin. It has long been assumed that a Golgi-specific protein 4.1 must also exist, but it has not previously been found. We demonstrate here that a hitherto unknown form of protein 4.1, a 200 kDa 4.1B, is associated with the Golgi of Madin-Darby canine kidney (MDCK) and human bronchial epithelial (HBE) cells. This 4.1B variant behaves like a Golgi marker after treatment with Brefeldin A and during mitosis. Depletion of the protein in HBE cells by siRNA resulted in disruption of the Golgi structure and failure of Na(+)/K(+)-ATPase, ZO-1 and ZO-2 to migrate to the membrane. Thus, this newly identified Golgi-specific protein 4.1 appears to have an essential role in maintaining the structure of the Golgi and in assembly of a subset of membrane proteins. PMID- 19299465 TI - S. pombe btn1, the orthologue of the Batten disease gene CLN3, is required for vacuole protein sorting of Cpy1p and Golgi exit of Vps10p. AB - Batten disease is characterised by lysosomal dysfunction. The most common type of the disease is caused by mutations in the membrane protein CLN3, whose function is unknown. We show that the fission yeast orthologue Btn1p, previously implicated in vacuole function, is required for correct sorting of the vacuole hydrolase carboxypeptidase Y (Cpy1p). This is, in part, due to a defect in trafficking of Vps10p, the sorting receptor for Cpy1p, from the Golgi to the trans-Golgi network in btn1Delta cells. Our data also implicate btn1 in other Vps10-independent Cpy1-sorting pathways. Furthermore, btn1 affects the number, intracellular location and structure of Golgi compartments. We show that the prevacuole location of Btn1p is at the Golgi, because Btn1p colocalises predominantly with the Golgi marker Gms1p in compartments that are sensitive to Brefeldin A. Btn1p function might be linked to that of Vps34p, a phosphatidylinositol 3-kinase, because Btn1p acts as a multicopy suppressor of the severe Cpy1p vacuole protein-sorting defect of vps34Delta cells. Together, these results indicate an important role for Btn1p in the Golgi complex, which affects Golgi homeostasis and vacuole protein sorting. We propose a similar role for CLN3 in mammalian cells. PMID- 19299466 TI - p300-mediated acetylation of the Rothmund-Thomson-syndrome gene product RECQL4 regulates its subcellular localization. AB - RECQL4 belongs to the conserved RecQ family of DNA helicases, members of which play important roles in the maintenance of genome stability in all organisms that have been examined. Although genetic alterations in the RECQL4 gene are reported to be associated with three autosomal recessive disorders (Rothmund-Thomson, RAPADILINO and Baller-Gerold syndromes), the molecular role of RECQL4 still remains poorly understood. Here, we show that RECQL4 specifically interacts with the histone acetyltransferase p300 (also known as p300 HAT), both in vivo and in vitro, and that p300 acetylates one or more of the lysine residues at positions 376, 380, 382, 385 and 386 of RECQL4. Furthermore, we report that these five lysine residues lie within a short motif of 30 amino acids that is essential for the nuclear localization of RECQL4. Remarkably, the acetylation of RECQL4 by p300 in vivo leads to a significant shift of a proportion of RECQL4 protein from the nucleus to the cytoplasm. This accumulation of the acetylated RECQL4 is a result of its inability to be imported into the nucleus. Our results provide the first evidence of a post-translational modification of the RECQL4 protein, and suggest that acetylation of RECQL4 by p300 regulates the trafficking of RECQL4 between the nucleus and the cytoplasm. PMID- 19299467 TI - Stability of the small gamma-tubulin complex requires HCA66, a protein of the centrosome and the nucleolus. AB - To investigate changes at the centrosome during the cell cycle, we analyzed the composition of the pericentriolar material from unsynchronized and S-phase arrested cells by gel electrophoresis and mass spectrometry. We identified HCA66, a protein that localizes to the centrosome from S-phase to mitosis and to the nucleolus throughout interphase. Silencing of HCA66 expression resulted in failure of centrosome duplication and in the formation of monopolar spindles, reminiscent of the phenotype observed after gamma-tubulin silencing. Immunofluorescence microscopy showed that proteins of the gamma-tubulin ring complex were absent from the centrosome in these monopolar spindles. Immunoblotting revealed reduced protein levels of all components of the gamma tubulin small complex (gamma-tubulin, GCP2, and GCP3) in HCA66-depleted cells. By contrast, the levels of gamma-tubulin ring complex proteins such as GCP4 and GCP WD/NEDD1 were unaffected. We propose that HCA66 is a novel regulator of gamma tubulin function that plays a role in stabilizing components of the gamma-tubulin small complex, which is in turn essential for assembling the larger gamma-tubulin ring complex. PMID- 19299468 TI - NDST1-dependent heparan sulfate regulates BMP signaling and internalization in lung development. AB - Heparan sulfate proteoglycans (HSPGs) are required for various signaling pathways, one of which is the bone morphogenetic protein (BMP) signaling pathway. N-deacetylase/N-sulfotransferase-1 (NDST1) participates in synthesizing heparan sulfate (HS) chains of HSPGs, and is involved in bone and lung development. Here, we report that in spite of the redundant expression of Ndst2, Ndst3 and Ndst4 genes, Ndst1(-/-) mice display defective differentiation of lung cells and increased cell proliferation. Loss of Ndst1 in the lung enhances downstream BMP signaling in vivo. Noggin, which is an antagonist of BMP, can rescue the Ndst1(-/ ) lung morphogenetic defects in explant cultures. Further studies in vitro indicated that loss of Ndst1 significantly impairs BMP internalization by decreasing BMP binding to endogenous HS. Exogenous heparin can rescue both the BMP signaling and BMP internalization abnormalities in Ndst1(-/-) lung. Thus, we propose that HS regulates BMP signaling by controlling the balance between BMP binding to HS, and that BMP receptors and NDST1-dependent modification are essential for this process. The results suggest that NDST1-dependent HS is essential for proper functioning of BMP in embryonic lung development. PMID- 19299469 TI - Eight years' experience of an extended-interval dosing protocol for gentamicin in neonates. AB - BACKGROUND: Dosing of gentamicin in neonates in Christchurch has been carried out since 2000 using a locally developed extended-interval dosing protocol. All dosing data have been recorded in a database. AIMS: The aims of this study were to analyse the database to determine what percentage of neonates achieved target values for C(max), C(min) and AUC, and to use the pharmacokinetic values of gentamicin to simulate new dosing protocols. METHODS: C(max), C(min) and AUC were compared with target values. Clearance (CL), volume of distribution (V) and half life (t(1/2)) were estimated, and used to produce new predictive dosing protocols that were tested and compared with the results of the original protocol. RESULTS: Gentamicin concentrations from 1461 individual doses were recorded in the database. Four hundred and eight were excluded. Of the remaining 1053, 84% achieved the target C(max) (>10 mg/L), 77% the target C(min) (<1 mg/L) and 63% the target AUC (within 80% to 125%). The number achieving target C(max) and C(min) values was improved markedly by prolonging the dosing intervals, but not by altering the predictive equations. Since the majority of the neonates only received a single dose of gentamicin, a new V-based model was also tested, and performed well. CL (L/kg) increased, while V (L/kg) and t(1/2) (h) both decreased with respect to weight. CONCLUSIONS: Extending the dose interval improved the success in achieving target C(max) and C(min), while revision of the dosing equation did not. A V-based model provides an alternative approach to the first dose of gentamicin in neonates. PMID- 19299470 TI - Comment on: Resistance of planktonic and biofilm-grown Burkholderia cepacia complex isolates to the transition metal gallium. PMID- 19299472 TI - Development and evaluation of vancomycin dosage guidelines designed to achieve new target concentrations. AB - OBJECTIVES: The aims of this study were to develop a population pharmacokinetic model of vancomycin in adult patients, to use this model to develop dosage guidelines targeting vancomycin trough concentrations of 10-15 mg/L and to evaluate the performance of these new guidelines. METHODS: All data analyses were performed using NONMEM. A population pharmacokinetic model was first developed from vancomycin dosage and concentration data collected during routine therapeutic drug monitoring in 398 patients, then new vancomycin dosage guidelines were devised by using the model to predict vancomycin trough concentrations in a simulated dataset. Individual estimates of CL and V1 were then obtained in an independent group of 100 patients using the population model and the POSTHOC option. These individual estimates were used to predict vancomycin trough concentrations and steady-state AUC(24)/MIC ratios using the current and new dosage guidelines. RESULTS: The population analysis found that the vancomycin data were best described using a bi-exponential elimination model with a typical CL of 3.0 L/h that changed by 15.4% for every 10 mL/min difference from a CL(CR) of 66 mL/min. V(ss) was 1.4 L/kg. The proposed dosage guidelines were predicted to achieve 55% of vancomycin troughs within 10-15 mg/L and 71% within 10-20 mg/L, which is significantly higher than current guidelines (19% and 22%, respectively). The proportion of AUC(24)/MIC ratios above 400 was also higher, 87% compared with 58%. CONCLUSIONS: New vancomycin dosage guidelines have been developed that achieve trough concentrations of 10-15 mg/L earlier and more consistently than current guidelines. PMID- 19299471 TI - Peripheral and visceral fat changes following a treatment switch to a non thymidine analogue or a nucleoside-sparing regimen in HIV-infected subjects with peripheral lipoatrophy: results of ACTG A5110. AB - BACKGROUND: Switching a thymidine analogue to a non-thymidine analogue or changing to a nucleoside-sparing regimen has been shown to partially reverse peripheral lipoatrophy. The current study evaluated both approaches. METHODS: Subjects at 15 AIDS Clinical Trial Group sites receiving thymidine analogue stavudine- or zidovudine-containing regimens with plasma HIV RNA < or =500 copies/mL and lipoatrophy were prospectively randomized to: (i) switch the thymidine analogue to abacavir; (ii) discontinue all antiretrovirals and switch to lopinavir/ritonavir plus nevirapine (LPV/r+NVP); or (iii) delay switching for 24 weeks (ClinicalTrials.gov identifier: NCT00028314). Single-slice computer tomography of mid-thigh and abdominal fat and metabolic and virological/immunological parameters were measured at baseline and weeks 24 and 48. RESULTS: Among the 101 patients enrolled, there were significant subcutaneous thigh fat and subcutaneous abdominal tissue (SAT) increases over time and decreases in visceral adipose tissue to total adipose tissue (VAT:TAT) ratios for both interventions, and a decrease in VAT for abacavir. CD4 increased in the LPV/r+NVP arm. LPV/r+NVP had a significantly shorter time to grade 3 or higher toxicity (P = 0.007), but discontinuation rates were similar. Glucose levels did not change, but insulin decreased in the LPV/r+NVP arm. Lipids tended to increase in the LPV/r+NVP arm. CONCLUSIONS: Switching stavudine or zidovudine to a non thymidine analogue or changing to a nucleoside reverse transcriptase inhibitor sparing regimen is associated with qualitatively similar improvements in thigh fat, SAT and VAT:TAT ratio at 48 weeks. Abacavir also resulted in VAT reductions and LPV/r+NVP resulted in CD4 count increases. PMID- 19299473 TI - StaphVar-DNA microarray analysis of accessory genome elements of community acquired methicillin-resistant Staphylococcus aureus. AB - OBJECTIVES: Approximately 75% of the genome of Staphylococcus aureus (the 'core' genome) is highly conserved between strains, whereas the remaining 25% (the 'accessory' genome) is composed of mobile genetic elements (MGEs), containing virulence and resistance genes. We developed a composite microarray focused on resistance and virulence genes located on the accessory or core-variable genome to characterize a collection of Belgian community-acquired methicillin-resistant S. aureus (CA-MRSA) strains. METHODS: Oligonucleotide probes targeting 403 genes encoding antimicrobial resistance (35%), virulence (28%) and adhesion (31%) factors were designed among eight S. aureus sequenced genomes. The StaphVar Array was validated by testing five of the strains used for the design and utilized to characterize 13 CA-MRSA strains representative of the multilocus sequence typing (MLST) sequence types circulating in Belgium. RESULTS: Analysis of the gene content of the five reference strains by the StaphVar Array matched 90% to 97% of the theoretical results. Analysis of CA-MRSA strains showed that 54.4% of the genes tested were strain-dependent. Strains presented specific exotoxin, enterotoxin, cytolysin and adhesin gene profiles by MLST lineage. One exception to these 'lineage-specific' profiles was the variable presence of the arginine catabolic mobile element (characteristic of the USA300 clone) within ST8 strains. CONCLUSIONS: The StaphVar Array enables the characterization of approximately 400 variable resistance and virulence determinants in S. aureus. CA-MRSA strains displayed extensive diversity in virulence and resistance profiles. The presence of the USA300 clone in Belgium was confirmed. Although mainly located on MGEs, associations of virulence genes were highly conserved within strains of the same MLST lineage. PMID- 19299474 TI - Patients' and doctors' views on depression severity questionnaires incentivised in UK quality and outcomes framework: qualitative study. AB - OBJECTIVE: To gain understanding of general practitioners' and patients' opinions of the routine introduction of standardised measures of severity of depression through the UK general practice quality and outcomes framework. DESIGN: Semistructured qualitative interview study, with purposive sampling and constant comparative analysis. PARTICIPANTS: 34 general practitioners and 24 patients. SETTING: 38 general practices in three sites in England: Southampton, Liverpool, and Norfolk. RESULTS: Patients generally favoured the measures of severity for depression, whereas general practitioners were generally cautious about the validity and utility of such measures and sceptical about the motives behind their introduction. Both general practitioners and patients considered that assessments of severity should be seen as one aspect of holistic care. General practitioners considered their practical wisdom and clinical judgment ("phronesis") to be more important than objective assessments and were concerned that the assessments reduced the human element of the consultation. Patients were more positive about the questionnaires, seeing them as an efficient and structured supplement to medical judgment and as evidence that general practitioners were taking their problems seriously through a full assessment. General practitioners and patients were aware of the potential for manipulation of indicators: for economic reasons for doctors and for patients to avoid stigma or achieve desired outcomes. CONCLUSIONS: Despite general practitioners' caution about measures of severity for depression, these may benefit primary care consultations by increasing patients' confidence that general practitioners are correct in their diagnosis and are making systematic efforts to assess and manage their mental health problems. Further education of primary care staff may optimise the use and interpretation of depression questionnaires. PMID- 19299476 TI - Treatment of depression in primary care. PMID- 19299475 TI - Management of depression in UK general practice in relation to scores on depression severity questionnaires: analysis of medical record data. AB - OBJECTIVE: To determine if general practitioner rates of antidepressant drug prescribing and referrals to specialist services for depression vary in line with patients' scores on depression severity questionnaires. DESIGN: Analysis of anonymised medical record data. SETTING: 38 general practices in three sites Southampton, Liverpool, and Norfolk. Data reviewed Records for 2294 patients assessed with severity questionnaires for depression between April 2006 and March 2007 inclusive. MAIN OUTCOME MEASURES: Rates of prescribing of antidepressants and referrals to specialist mental health or social services. RESULTS: 1658 patients were assessed with the 9 item patient health questionnaire (PHQ-9), 584 with the depression subscale of the hospital anxiety and depression scale (HADS), and 52 with the Beck depression inventory, 2nd edition (BDI-II). Overall, 79.1% of patients assessed with either PHQ-9 or HADS received a prescription for an antidepressant, and 22.8% were referred to specialist services. Prescriptions and referrals were significantly associated with higher severity scores. However, overall rates of treatment and referral were similar for patients assessed with either measure despite the fact that, with PHQ-9, 83.5% of patients were classified as moderately to severely depressed and in need of treatment, whereas only 55.6% of patients were so classified with HADS. Rates of treatment were lower for older patients and for patients with comorbid physical illness (including coronary heart disease and diabetes) despite the fact that screening for depression among such patients is encouraged in the quality and outcomes framework. Conclusions General practitioners do not decide on drug treatment or referral for depression on the basis of questionnaire scores alone, but also take account of other factors such as age and physical illness. The two most widely used severity questionnaires perform inconsistently in practice, suggesting that changing the recommended threshold scores for intervention might make the measures more valid, more consistent with practitioners' clinical judgment, and more acceptable to practitioners as a way of classifying patients. PMID- 19299477 TI - International Internal Dosimetry Network. PMID- 19299478 TI - Malaysian participation in the IAEA/WHO postal TLD and postal ionisation chamber intercomparison programmes: analysis of results obtained during 1985-2008. AB - During the years 1985-2008, the Secondary Standards Dosimetry Laboratory of Malaysia (SSDL Malaysia) has participated 37 times in the IAEA/WHO intercomparison programmes. This paper reports an analysis of the intercomparison data and demonstrates that the quality of the SSDL calibration service is well within the limits required by IAEA. PMID- 19299479 TI - The investigation of fetal doses in mantle field irradiation. AB - To determine clinically the fetal dose from irradiation of Hodgkin's disease during pregnancy and to quantify the components of fetal dose using phantom measurements. The fetal dose was measured with phantom measurements using thermoluminescent dosemeters (TLDs). Phantom measurements were performed by simulating the treatment conditions on an anthropomorphic phantom. TLDs were placed on the phantom 41, 44, 46.5 and 49.5 cm from the centre of the treatment field. Two TLDs were placed on the surface of the phantom. The estimated total dose to all the TLDs ranged from 8.8 to 13.2 cGy for treatment with (60)Co and from 8.2 to 11.8 cGy for 4 MV photons. It was concluded that the doses in different sections were evaluated to investigate dose changes in different points and depths of fetal tissues in phantom. Precise planning and the use of supplemental fetal shielding may help reduce fetal exposure. PMID- 19299480 TI - Cell-type specific activation of p38 and ERK mediates calcitonin gene-related peptide involvement in tolerance to morphine-induced analgesia. AB - Tolerance to morphine-induced analgesia is a well-established phenomenon, often limiting its usefulness in the long-term treatment of pain. The mechanisms underlying tolerance are not well understood. We previously suggested a possible role for spinal calcitonin gene-related peptide (CGRP) in the development of tolerance to morphine-induced analgesia. In the present study, we demonstrate that CGRP is involved in morphine tolerance by differentially regulating the ERK dependent up-regulation of IL-1beta, TNF-alpha, and microsomal prostaglandin E synthase-1 (mPGES-1) in astrocytes and p38-dependent up-regulation of IL-6 in microglia in the rat spinal cord. A 7-d treatment with morphine induced tolerance to the antinociceptive effect and increased phosphorylated ERK localized in astrocytes and phosphorylated p38 enriched in microglia, both effects being inhibited by blocking CGRP receptors. Interestingly, the inhibition of the ERK pathway suppressed the development of tolerance and morphine-induced up regulation of IL-1beta, TNF-alpha, and mPGES-1. Blockade of p38 activity also inhibited the development of tolerance and morphine-induced IL-6 up-regulation. Taken together, these data suggest that chronic morphine induces the synthesis of CGRP, which in turn acts on CGRP receptors located on astrocytes and microglia to stimulate ERK and p38, respectively, leading to increased synthesis and release of proinflammatory mediators resulting in tolerance to morphine-induced analgesia. PMID- 19299481 TI - Origin of the adult intestinal stem cells induced by thyroid hormone in Xenopus laevis. AB - In the amphibian intestine during metamorphosis, de novo stem cells generate the adult epithelium analogous to the mammalian counterpart. Interestingly, to date the exact origin of these stem cells remains to be determined, making intestinal metamorphosis a unique model to study development of adult organ-specific stem cells. Here, to determine their origin, we made use of transgenic Xenopus tadpoles expressing green fluorescent protein (GFP) for recombinant organ cultures. The larval epithelium separated from the wild-type (Wt) or GFP transgenic (Tg) intestine before metamorphic climax was recombined with homologous and heterologous nonepithelial tissues and was cultivated in the presence of thyroid hormone, the causative agent of metamorphosis. In all kinds of recombinant intestine, adult progenitor cells expressing markers for intestinal stem cells such as sonic hedgehog became detectable and then differentiated into the adult epithelium expressing intestinal fatty acid binding protein, a marker for absorptive cells. Notably, whenever the epithelium was derived from Tg intestine, both the adult progenitor/stem cells and their differentiated cells expressed GFP, whereas neither of them expressed GFP in the Wt-derived epithelium. Our results provide direct evidence that stem cells that generate the adult intestinal epithelium originate from the larval epithelium, through thyroid hormone-induced dedifferentiation. PMID- 19299482 TI - Neuronal palmitoyl acyl transferases exhibit distinct substrate specificity. AB - Palmitoylation, a post-translational modification of cysteine residues with the lipid palmitate, has recently emerged as an important mechanism for regulating protein trafficking and function. With the identification of 23 DHHC mammalian palmitoyl acyl transferases (PATs), a key question was the nature of substrate enzyme specificity for these PATs. Using the acyl-biotin exchange palmitoylation assay, we compared the substrate specificity of four neuronal PATs, namely DHHC 3, DHHC-8, HIP14L (DHHC-13), and HIP14 (DHHC-17). Exogenous expression of enzymes and substrates in COS cells reveals that HIP14L and HIP14 modulate huntingtin palmitoylation, DHHC-8 modulates paralemmin-1 palmitoylation, and DHHC-3 shows the least substrate specificity. These in vitro data were validated by lentiviral siRNA-mediated knockdown of endogenous HIP14 and DHHC-3 in cultured rat cortical neurons. PATs require the presence of palmitoylated cysteines in order to interact with their substrates. To understand the elements that influence enzyme/substrate specificity further, we fused the HIP14 ankryin repeat domain to the N terminus of DHHC-3, which is not a PAT for huntingtin. This modification enabled DHHC-3 to behave similarly to HIP14 by modulating palmitoylation and trafficking of huntingtin. Taken together, this study indicates that individual PATs have specific substrate preference, determined by regulatory domains outside the DHHC domain of the enzymes. PMID- 19299483 TI - 15-Deoxy-Delta12,14-prostaglandin J2 inhibits HIV-1 transactivating protein, Tat, through covalent modification. AB - Controlling the HIV/AIDS epidemic remains a major challenge, with approximately 5 million new HIV infections annually. Cyclopentenone prostaglandins (CyPG), such as 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)), are arachidonic acid-derived endogenous electrophiles that possess anti-HIV activity by an unknown mechanism. Given that the reactive alpha,beta-unsaturated ketone in the cyclopentenone ring of 15d-PGJ(2) covalently modifies key Cys thiols in select proteins, we hypothesized that 15d-PGJ(2) inhibits HIV transcription and replication by targeting Cys thiols in HIV-1 Tat. Tat is a potent transactivator of viral gene expression required for HIV transcriptional elongation and replication. Our studies indicate that 15d-PGJ(2) treatment of cells inhibits Tat-dependent transcription and replication of HIV-1, while 9,10-dihydro-15d-PGJ(2), PGE(2), PGF(2alpha), or PGD(2) that lack the reactive alpha,beta-unsaturated ketone were ineffective. The inhibition of Tat activity by 15d-PGJ(2) was dose-dependent, with an IC(50) of 1.2 microM and independent of NF-kappaB pathway. Furthermore, using a biotinylated derivative of 15d-PGJ(2), we demonstrate that 15d-PGJ(2) modifies free Cys-thiols in Tat to form covalent Michael adducts and that the interaction was further increased on reduction of Tat. 15d-PGJ(2)-modified Tat was unable to transactivate the HIV long terminal repeat in U937 human macrophages. These data demonstrate that Tat acts as a molecular target of CyPG leading to the inhibition of transcription and also suggest a novel therapeutic approach to complement current antiretroviral strategies for HIV/AIDS. PMID- 19299484 TI - Computerized assessment of angiographic occlusion rate and coil density in embolized human cerebral aneurysms. AB - BACKGROUND AND PURPOSE: Computerized methods have been introduced for more objective quantification of angiographic occlusion rate and coil density as parameters of successful embolization. This study aimed 1) to evaluate this new computerized method for angiographic occlusion rating and coil density calculations by comparison with corresponding histometric parameters from retrieved human aneurysms, and 2) to compare the new computerized method with the present standard of subjective angiographic occlusion rating. MATERIALS AND METHODS: From 14 postmortem-retrieved human aneurysms, angiographic occlusion rate was determined by contrast medium attenuation-gradient distinction on digital subtraction angiographs after Guglielmi detachable coil (GDC) embolization. Angiographic coil density was calculated, approximating aneurysms as ellipsoid and coils as cylindric volumes. On surface-stained histologic ground sections of the respective aneurysms, the occluded aneurysm area and coil area were measured. Then, we calculated and compared the histometric occlusion rates and coil densities with the corresponding angiographic parameters by using the Wilcoxon paired signed-rank test and the Spearman rank correlation. RESULTS: Computerized angiographic occlusion rates (75%-100%) showed good correlation (r = 0.799; P < .01) with histometric occlusion-rates (61%-100%), resulting in no statistically significant differences (P = .2163). With 5.1% (+/-3.8), the mean difference between computerized angiographic occlusion rates and histometry was substantially lower compared with 10.7% (+/-8.7) mean difference between subjective angiographic estimations and histometry. Calculated angiographic coil density (13%-32%) significantly differed from histometric coil density (8%-35%; P < .05). CONCLUSIONS: For recanalized aneurysms, computerized angiographic occlusion rating showed better correspondence with histometry compared with subjective angiographic occlusion rating. Clinical application of this new tool may lead to more objective cutoff values for re-embolization indications. The value of coil density calculations seems limited by the approximation of the aneurysms as ellipsoid volumes. PMID- 19299485 TI - Clinical and angiographic follow-up of ruptured intracranial aneurysms treated with endovascular embolization. AB - BACKGROUND AND PURPOSE: Endovascular embolization is a well-established treatment of ruptured intracranial aneurysms, but concern about its long-term stability and its ability to prevent rehemorrhage are still present. We evaluated the long-term clinical and angiographic follow-up of patients with ruptured cerebral aneurysms treated with coiling, focusing on rehemorrhage and changes in aneurysm morphologic features. MATERIALS AND METHODS: A total of 377 patients with ruptured aneurysms that were treated with endovascular approaches at our institution between 1994 and 2008 were reviewed. Clinical and angiographic data were analyzed from a prospectively collected data base. RESULTS: There were 377 patients with 391 ruptured aneurysms treated for 14 years. Good outcome (Glasgow Outcome Score [GOS], 5) was achieved in 74% of patients, moderate disability or poor outcome in 18%, and 8.8% died. Permanent morbidity or mortality from procedural complications occurred in 2.9%. Complete follow-up was available for 85% of surviving patients, with mean follow-up of 22.3 months. Re-treatment was required in 11% (31 patients). Eight (2.1%) patients had rebleeding, 6 (1.6%) in the hospital within 30 days of treatment, 5 in the first 48 hours. Follow-up imaging was available in 276 aneurysms in 270 patients. Recanalization occurred in 56 of 276 aneurysms (20.3%) regardless of the initial angiographic result, but the risk was higher if a body remnant was left (chi2, 11.791; P = .0006). CONCLUSIONS: Long-term clinical and angiographic follow-up demonstrates the efficacy of endovascular treatment of ruptured intracranial aneurysms. Rebleeding after treatment is rare, with the greatest risk during the first 48 hours after treatment. Initial angiographic results are not a useful predictor of clinical outcome or rehemorrhage. PMID- 19299486 TI - Characterization of CSF hydrodynamics in the presence and absence of tonsillar ectopia by means of computational flow analysis. AB - BACKGROUND AND PURPOSE: Phase-contrast MR imaging (PCMR) has only partially characterized cyclic CSF flow and pressure, which, hypothetically, have a role in the pathogenesis of syrinx and symptoms in the Chiari I malformation. Our goal was to use computational flow analysis (CFA) to better understand CSF hydrodynamics. MATERIALS AND METHODS: High-resolution MR images were obtained in a healthy volunteer and a patient with Chiari I malformation. With standard segmentation and discretization techniques, 3D models of the subarachnoid space, cerebellum, and spinal canal were created. CSF flow during systole and diastole were simulated with the boundary element method in the models. CSF velocities and pressures computed in the patient with Chiari I malformation were compared with those in the healthy volunteer. Flow patterns were also compared with PCMR results for validation of the technique. RESULTS: The CFA and PCMR results agreed well. Inhomogeneous flow patterns characterized by fluid jets anterior and lateral to the spinal cord were demonstrated in both the Chiari I and volunteer models by CFA. Significant circumferential velocities were evident, suggesting swirling flow in the spinal canal. Higher magnitude jets were found in the patient with Chiari I than in the healthy volunteer. Relatively even pressure gradients were found along the spinal canal in both cases, with a 50% steeper gradient in the patient with Chiari I malformation. CONCLUSIONS: Circumferential velocities and pressure gradients in the spinal canal, which may be clinically relevant to Chiari I and other malformations, can be obtained by CFA in patient specific geometries. PMID- 19299487 TI - Window settings for the study of calcified carotid plaques with multidetector CT angiography. AB - SUMMARY: CT angiography (CTA) shows high sensitivity in detecting calcified plaques, but sometimes a bias in the exact quantification of stenosis degree occurs, mainly caused by the high linear attenuation coefficient of the calcified plaques. The purpose of this technical study was to evaluate the most appropriate CT window parameters for the assessment of calcified plaques stating which of them can provide the best inter-observer agreement. Scatter-plots and regression results showed the correlation between both width and level respectively depending on intraluminal Hounsfield units (HU) value (width = intraluminal HU x 2.07; level = intraluminal HU x 0.72). Obtained data indicated that the presence of different stenosis degrees did not modify visualization parameters. PMID- 19299488 TI - C-arm CT measurement of cerebral blood volume: an experimental study in canines. AB - BACKGROUND AND PURPOSE: Cerebral blood volume (CBV) is an important parameter in estimating the viability of brain tissue following an ischemic event. We tested the hypothesis that C-arm CT measurements of CBV would correlate well with those made with perfusion CT (PCT). MATERIALS AND METHODS: CBV was measured in 12 canines by using PCT and C-arm CT. Two measurements with each technique were made on each animal; a different injection protocol was used for each of these techniques. PCT was performed by using a 64-section V-scanner. C-arm CT was performed by using a biplane Artis dBA system. PCT images were transferred to a commercially available workstation for postprocessing and analysis; C-arm CT images were transferred to a commercially available workstation for postprocessing and analysis by using prototype software. From each animal, 2 sections from each technique were selected for analysis. RESULTS: There was good agreement of both the color maps and absolute numbers between the 2 techniques. The maximum and mean deviations of values between the 2 techniques for the first 5 animals were 30.20% and 7.82%; for the second 7 animals, these values were 26.79% and 7.40%. The maximum and mean deviations between the 2 C-arm CT studies performed on the first 5 animals were 33.15% and 12.24%; for the second 7 animals, these values were 41.15% and 10.89%. CONCLUSIONS: In these healthy animals, measurement of CBV with C-arm CT compared well with measurements made with PCT. PMID- 19299489 TI - Theoretic basis and technical implementations of CT perfusion in acute ischemic stroke, part 2: technical implementations. AB - CT perfusion (CTP) is a functional imaging technique that provides important information about capillary-level hemodynamics of the brain parenchyma and is a natural complement to the strengths of unenhanced CT and CT angiography in the evaluation of acute stroke, vasospasm, and other neurovascular disorders. CTP is critical in determining the extent of irreversibly infarcted brain tissue (infarct "core") and the severely ischemic but potentially salvageable tissue ("penumbra"). This is achieved by generating parametric maps of cerebral blood flow, cerebral blood volume, and mean transit time. PMID- 19299490 TI - Hypophyseal triplication: case report and embryologic considerations. AB - SUMMARY: Hypophyseal triplication is malformation that has not been described previously. We present a child with midline abnormalities who underwent epignathus excision at birth. Brain MR imaging revealed 2 paired lateral pituitary glands and an oval midline gland, each with an independent stalk, connected to a thickened third ventricle floor. Because malformations represent a failure in embryogenesis, this case may provide interesting clues on the normal development of the hypophysis. PMID- 19299491 TI - Percutaneous transluminal angioplasty and stent placement in acute vessel occlusion: evaluation of new methods for interventional stroke treatment. AB - BACKGROUND AND PURPOSE: The major goal of acute ischemic stroke treatment is fast and sufficient recanalization. Percutaneous transluminal balloon angioplasty (PTA) and/or placement of a stent might achieve both by compressing the thrombus at the occlusion site. This study assesses the feasibility, recanalization rate, and complications of the 2 techniques in an animal model. MATERIALS AND METHODS: Thirty cranial vessels of 7 swine were occluded by injection of radiopaque thrombi. Fifteen vessel occlusions were treated by PTA alone and 15, by placement of a stent and postdilation. Recanalization was documented immediately after treatment and after 1, 2, and 3 hours. Thromboembolic events and dissections were documented. RESULTS: PTA was significantly faster to perform (mean, 16.6 minutes versus 33.0 minutes for stent placement; P < .001), but the mean recanalization rate after 1 hour was significantly better after stent placement compared with PTA alone (67.5% versus 14.6%, P < .001). Due to the self-expanding force of the stent, vessel diameter further increased with time, whereas the recanalization result after PTA was prone to reocclusion. Besides thromboembolic events related to the passing maneuvers at the occlusion site, no thrombus fragmentation and embolization occurred during balloon inflation or stent deployment. Flow to side branches could also be restored at the occlusion site because it was possible to direct thrombus compression. CONCLUSIONS: Stent placement and postdilation proved to be much more efficient in terms of acute and short-term vessel recanalization compared with PTA alone. PMID- 19299492 TI - Gene therapy for retinitis pigmentosa and Leber congenital amaurosis caused by defects in AIPL1: effective rescue of mouse models of partial and complete Aipl1 deficiency using AAV2/2 and AAV2/8 vectors. AB - Defects in the photoreceptor-specific gene encoding aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) are clinically heterogeneous and present as Leber Congenital Amaurosis, the severest form of early-onset retinal dystrophy and milder forms of retinal dystrophies such as juvenile retinitis pigmentosa and dominant cone-rod dystrophy. [Perrault, I., Rozet, J.M., Gerber, S., Ghazi, I., Leowski, C., Ducroq, D., Souied, E., Dufier, J.L., Munnich, A. and Kaplan, J. (1999) Leber congenital amaurosis. Mol. Genet. Metab., 68, 200-208.] Although not yet fully elucidated, AIPL1 is likely to function as a specialized chaperone for rod phosphodiesterase (PDE). We evaluate whether AAV-mediated gene replacement therapy is able to improve photoreceptor function and survival in retinal degeneration associated with AIPL1 defects. We used two mouse models of AIPL1 deficiency simulating three different rates of photoreceptor degeneration. The Aipl1 hypomorphic (h/h) mouse has reduced Aipl1 levels and a relatively slow degeneration. Under light acceleration, the rate of degeneration in the Aipl1 h/h mouse is increased by 2-3-fold. The Aipl1-/- mouse has no functional Aipl1 and has a very rapid retinal degeneration. To treat the different rates of degeneration, two pseudotypes of recombinant adeno-associated virus (AAV) exhibiting different transduction kinetics are used for gene transfer. We demonstrate restoration of cellular function and preservation of photoreceptor cells and retinal function in Aipl1 h/h mice following gene replacement therapy using an AAV2/2 vector and in the light accelerated Aipl1 h/h model and Aipl1-/- mice using an AAV2/8 vector. We have thus established the potential of gene replacement therapy in varying rates of degeneration that reflect the clinical spectrum of disease. This is the first gene replacement study to report long-term rescue of a photoreceptor-specific defect and to demonstrate effective rescue of a rapid photoreceptor degeneration. PMID- 19299493 TI - Biochemical and genetic evidence for a role of IGHMBP2 in the translational machinery. AB - The human motor neuron degenerative disease spinal muscular atrophy with respiratory distress type 1 (SMARD1) is caused by loss of function mutations of immunoglobulin mu-binding protein 2 (IGHMBP2), a protein of unknown function that contains DNA/RNA helicase and nucleic acid-binding domains. Reduced IGHMBP2 protein levels in neuromuscular degeneration (nmd) mice, the mouse model of SMARD1, lead to motor neuron degeneration. We report the biochemical characterization of IGHMBP2 and the isolation of a modifier locus that rescues the phenotype and motor neuron degeneration of nmd mice. We find that a 166 kb BAC transgene derived from CAST/EiJ mice and containing tRNA genes and activator of basal transcription 1 (Abt1), a protein-coding gene that is required for ribosome biogenesis, contains the genetic modifier responsible for motor neuron rescue. Our biochemical investigations show that IGHMBP2 associates physically with tRNAs and in particular with tRNA(Tyr), which are present in the modifier and with the ABT1 protein. We find that transcription factor IIIC-220 kDa (TFIIIC220), an essential factor required for tRNA transcription, and the helicases Reptin and Pontin, which function in transcription and in ribosome biogenesis, are also part of IGHMBP2-containing complexes. Our findings strongly suggest that IGHMBP2 is a component of the translational machinery and that these components can be manipulated genetically to suppress motor neuron degeneration. PMID- 19299494 TI - Distinct mutations in the glycogen debranching enzyme found in glycogen storage disease type III lead to impairment in diverse cellular functions. AB - Glycogen storage disease type III (GSDIII) is a metabolic disorder characterized by a deficiency in the glycogen debranching enzyme, amylo-1,6-glucosidase,4-alpha glucanotransferase (AGL). Patients with GSDIII commonly exhibit hypoglycemia, along with variable organ dysfunction of the liver, muscle or heart tissues. The AGL protein binds to glycogen through its C-terminal region, and possesses two separate domains for the transferase and glucosidase activities. Most causative mutations are nonsense, and how they affect the enzyme is not well understood. Here we investigated four rare missense mutations to determine the molecular basis of how they affect AGL function leading to GSDIII. The L620P mutant primarily abolishes transferase activity while the R1147G variant impairs glucosidase function. Interestingly, mutations in the carbohydrate-binding domain (CBD; G1448R and Y1445ins) are more severe in nature, leading to significant loss of all enzymatic activities and carbohydrate binding ability, as well as enhancing targeting for proteasomal degradation. This region (Y1445-G1448R) displays virtual identity across human and bacterial species, suggesting an important role that has been conserved throughout evolution. Our results clearly indicate that inactivation of either enzymatic activity is sufficient to cause GSDIII disease and suggest that the CBD of AGL plays a major role to coordinate its functions and regulation by the ubiquitin-proteasome system. PMID- 19299495 TI - Costs and cost-effectiveness of four treatment regimens for latent tuberculosis infection. AB - RATIONALE: Isoniazid given daily for 9 months is the standard treatment for latent tuberculosis infection (LTBI), but its effectiveness is limited by poor completion rates. Shorter course regimens and regimens using directly observed therapy result in improved adherence but have higher upfront costs. OBJECTIVES: To evaluate the costs and cost-effectiveness of regimens for the treatment of LTBI. METHODS: We used a computerized Markov model to estimate total societal costs and benefits associated with four regimens for the treatment of LTBI: self administered isoniazid daily for 9 months, directly observed isoniazid twice weekly for 9 months, directly observed isoniazid plus rifapentine once weekly for 3 months, and self-administered rifampin daily for 4 months. In the base-case analysis, subjects were assumed to have newly positive tuberculin skin tests after recent exposure to infectious tuberculosis. MEASUREMENTS AND MAIN RESULTS: We determined the costs of treatment, quality-adjusted life-years gained, and cases of active tuberculosis prevented. In the base-case analysis, rifampin dominated (less costly with increased benefits) all other regimens except isoniazid plus rifapentine, which was more effective at a cost $48,997 per quality-adjusted life year gained. Isoniazid plus rifapentine dominated all regimens at a relative risk of disease 5.2 times the baseline estimate, or with completion rates less than 34% for isoniazid or 37% for rifampin. Rifampin could be 17% less efficacious than self-administered isoniazid and still be cost-saving compared with this regimen. CONCLUSIONS: In our model, rifampin is cost-saving compared with the standard therapy of self-administered isoniazid. Isoniazid plus rifapentine is cost-saving for extremely high-risk patients and is cost-effective for lower-risk patients. PMID- 19299496 TI - Airway delivery of low-dose miglustat normalizes nasal potential difference in F508del cystic fibrosis mice. AB - RATIONALE: N-butyldeoxynojyrimicin (NB-DNJ, miglustat [Zavesca]) an approved drug for treating Gaucher disease, was reported to be able to correct the defective trafficking of the F508del-CFTR protein. OBJECTIVES: To evaluate the efficacy of in vivo airway delivery of miglustat for restoring ion transport in cystic fibrosis (CF). METHODS: We used nasal transepithelial potential difference (PD) as a measure of sodium and chloride transport. The effect of nasal instillation of a single dose of miglustat was investigated in F508del, cftr knockout and normal homozygous mice. The galactose iminosugar analog N butyldeoxygalactonojirimycin (NB-DGJ) was used as a placebo. MEASUREMENTS AND MAIN RESULTS: In F508del mice, sodium conductance (evaluated by basal hyperpolarization) and chloride conductance (evaluated by perfusing the nasal mucosa with chloride-free solution in the presence of amiloride and forskolin) were normalized 1 hour after an intranasal dose of 50 picomoles of miglustat. Chloride conductance in the presence of 200 microM 4-4'-diisothiocyanostilbene 2,2'-disulphonic acid (DIDS), an inhibitor of alternative chloride channels, was much higher after miglustat than after placebo. In cftr knockout mice, a normalizing effect was observed on sodium but not on chloride conductance. CONCLUSIONS: Our results provide clear evidence that nasal delivery of miglustat, at picomolar doses, normalizes sodium and Cftr-dependent chloride transport in F508del transgenic mice; they highlight the potential of topical miglustat as a therapy for CF. PMID- 19299497 TI - Heliox improves oxygen delivery and utilization during dynamic exercise in patients with chronic obstructive pulmonary disease. AB - RATIONALE: Normoxic heliox (mixture of 79% He and 21% O(2)) may enhance exercise tolerance in patients with chronic obstructive pulmonary disease (COPD). It remains to be determined whether part of these beneficial effects could be ascribed to increased O(2) delivery (O(2)DEL) to locomotor muscles. OBJECTIVES: To investigate the effects of heliox on peripheral O(2)DEL and utilization during exercise in moderate to severe COPD. METHODS: Twelve mildly hypoxic or nonhypoxemic men (FEV(1) = 45.0 +/- 13.0% predicted) underwent constant-work rate tests (70-80% peak) to the limit of tolerance while receiving heliox or room air. Near-infrared spectroscopy determined changes (Delta) in leg muscle deoxygenation (deoxyhemoglobin concentration [HHb], an index of fractional O(2) extraction), and surface electromyography estimated muscle fiber recruitment (n = 5). Q and Sp(O(2)) were monitored by impedance cardiography and pulse oximetry, respectively. MEASUREMENTS AND MAIN RESULTS: Heliox significantly decreased dynamic hyperinflation and increased exercise tolerance compared with room air (640 +/- 95 s vs. 371 +/- 100 s; P < 0.01). Heliox also accelerated on-exercise dynamics of Q, which were accompanied by faster O(2) uptake kinetics and slower Delta[HHb] responses (P < 0.05). During steady-state exercise, Sp(O(2))-corrected Delta[HHb] values decreased with heliox despite no significant changes in cardiac output. Muscle fiber recruitment and leg effort scores were also diminished (P < 0.05). On a multiple regression analysis, reductions in dynamic hyperinflation, dyspnea, and Delta[HHb] were independently related to improvements in exercise tolerance with heliox (R(2) = 0.91; P < 0.01). CONCLUSIONS: Heliox increases lower limb O(2)DEL and utilization during dynamic exercise in patients with moderate to severe COPD. These effects enhance exercise tolerance in this patient population. PMID- 19299498 TI - Clinical impact and reliability of pleural fluid mesothelin in undiagnosed pleural effusions. AB - RATIONALE: Serum mesothelin is a new biomarker for the diagnosis of mesothelioma. Patients with mesothelioma commonly present with pleural effusions. To define the clinical utility of mesothelin quantification in pleural fluid, we assessed its additional value over pleural fluid cytology and its short-term reproducibility and reliability after pleural inflammatory processes, including pleurodesis. OBJECTIVES: To assess the diagnostic role of pleural fluid mesothelin and the effect of common clinical factors that may influence measurement accuracy. METHODS: Mesothelin was quantified in 424 pleural fluid and 64 serum samples by ELISA. Fluid was collected prospectively from 167 patients who presented with pleural effusions for investigation. Serial pleural fluid samples were obtained from patients (n = 33) requiring repeated drainage. Mesothelin levels were also measured in patients (n = 32) prepleurodesis and postpleurodesis. MEASUREMENTS AND MAIN RESULTS: Pleural fluid mesothelin concentrations were significantly higher in patients with mesothelioma (n = 24) relative to those with metastatic carcinomas (n = 67) and benign effusions (n = 75): median (interquartile range, 25th-75th percentile) = 40.3 (18.3-68.1) versus 6.1 (1.5-13.2) versus 3.7 (0.0 12.4) nM, respectively, P < 0.0001. Mesothelin measurement was superior to cytological examination in the diagnosis and exclusion of mesothelioma (sensitivity, 71 vs. 35%; specificity, 89 vs. 100%; negative predictive value, 95 vs. 82%, respectively). In patients with "suspicious" cytology, pleural fluid mesothelin was 100% specific for mesothelioma, and in cytology-negative effusions (n = 105) offered a negative predictive value of 94%. Intraindividual reproducibility of pleural fluid mesothelin was excellent: mean (+/-SD) variation, -0.15 (+/-8.41) nM in samples collected within 7 days from patients with mesothelioma. Measurements remained reliable after pleurodesis and were not affected by the presence of bacteria. CONCLUSIONS: Pleural fluid mesothelin provides additional diagnostic value relative to cytological examination. Mesothelin measurements are reproducible and not affected by inflammatory pleural processes. PMID- 19299499 TI - Hospital admissions and chemical composition of fine particle air pollution. AB - RATIONALE: There are unexplained geographical and seasonal differences in the short-term effects of fine particulate matter (PM(2.5)) on human health. The hypothesis has been advanced to include the possibility that such differences might be due to variations in the PM(2.5) chemical composition, but evidence supporting this hypothesis is lacking. OBJECTIVES: To examine whether variation in the relative risks (RR) of hospitalization associated with ambient exposure to PM(2.5) total mass reflects differences in PM(2.5) chemical composition. METHODS: We linked two national datasets by county and by season: (1) long-term average concentrations of PM(2.5) chemical components for 2000-2005 and (2) RRs of cardiovascular and respiratory hospitalizations for persons 65 years or older associated with a 10-microg/m(3) increase in PM(2.5) total mass on the same day for 106 U.S. counties for 1999 through 2005. MEASUREMENTS AND MAIN RESULTS: We found a positive and statistically significant association between county specific estimates of the short-term effects of PM(2.5) on cardiovascular and respiratory hospitalizations and county-specific levels of vanadium, elemental carbon, or nickel PM(2.5) content. CONCLUSIONS: Communities with higher PM(2.5) content of nickel, vanadium, and elemental carbon and/or their related sources were found to have higher risk of hospitalizations associated with short-term exposure to PM(2.5). PMID- 19299500 TI - Pulmonary suppressor of cytokine signaling-1 induced by IL-13 regulates allergic asthma phenotype. AB - RATIONALE: Th2 cytokines play an important role in allergic diseases. These cytokines activate signal transduction pathways, including Janus kinase/signal transducer and activator of transcription (STAT) signaling. Although the suppressor of cytokine signaling (SOCS) family protein, a negative regulator of the Janus kinase/STAT signaling pathway, contributes to helper T cell differentiation during immune responses, the role of SOCS proteins within the structural cells of a target organ has not been clarified in allergy. OBJECTIVES: To study the local function of SOCS in the development of asthma. METHODS: We used mouse models of IL-13- and ovalbumin (OVA)-induced allergic airway disease. Airway smooth muscle cells were cultured from patients with asthma. MEASUREMENTS AND MAIN RESULTS: The administration of IL-13 induced not only airway responses but also SOCS1 expression at the local inflammatory site. The up-regulated SOCS1 markedly suppressed IL-13-dependent STAT6 activation and eotaxin expression and subsequently down-regulated IL-13-induced airway inflammatory responses. The inactivation of SOCS1 induced airway hyperresponsiveness after IL-13 treatment even in hyporesponsive C57BL/6 background mice. In an OVA-induced model of allergic airway disease, allergen exposure up-regulated local SOCS1 expression, and the induction of SOCS1 in the airways attenuated allergen-induced airway responses. Inactivation of IL-13 inhibited SOCS1 induction in a model of allergic airway disease. Interestingly, airway smooth muscle cells from individuals with asthma had impaired up-regulation of SOCS1 after IL-13 stimulation. CONCLUSIONS: SOCS1 induction by IL-13 in airway structural cells is critical to negatively control allergic airway disease. PMID- 19299501 TI - RhoA and Rho kinase activation in human pulmonary hypertension: role of 5-HT signaling. AB - RATIONALE: The complex and multifactorial pathogenesis of pulmonary hypertension (PH) involves constriction, remodeling, and in situ thrombosis of pulmonary vessels. Both serotonin (5-HT) and Rho kinase signaling may contribute to these alterations. OBJECTIVES: To investigate possible links between the 5-HT transporter (5-HTT) and RhoA/Rho kinase pathways, as well as their involvement in the progression of human and experimental PH. METHODS: Biochemical and functional analyses of lungs, platelets, and pulmonary artery smooth muscle cells (PA-SMCs) from patients with idiopathic PH (iPH) and 5-HTT overexpressing mice. MEASUREMENTS AND MAIN RESULTS: Lungs, platelets, and PA-SMCs from patients with iPH were characterized by marked elevation in RhoA and Rho kinase activities and a strong increase in 5-HT binding to RhoA indicating RhoA serotonylation. The 5 HTT inhibitor fluoxetine and the type 2 transglutaminase inhibitor monodansylcadaverin prevented 5-HT-induced RhoA serotonylation and RhoA/Rho kinase activation, as well as 5-HT-induced proliferation of PA-SMCs from iPH patients that was also inhibited by the Rho kinase inhibitor fasudil. Increased Rho kinase activity, RhoA activation, and RhoA serotonylation were also observed in lungs from SM22-5-HTT(+)mice, which overexpress 5-HTT in smooth muscle and spontaneously develop PH. Treatment of SM22-5-HTT(+) mice with either fasudil or fluoxetine limited PH progression and RhoA/Rho kinase activation. CONCLUSIONS: RhoA and Rho kinase activities are increased in iPH, in association with enhanced RhoA serotonylation. Direct involvement of the 5-HTT/RhoA/Rho kinase signaling pathway in 5-HT-mediated PA-SMC proliferation and platelet activation during PH progression identify RhoA/Rho kinase signaling as a promising target for new treatments against PH. PMID- 19299502 TI - The cost of gene expression underlies a fitness trade-off in yeast. AB - Natural selection optimizes an organism's genotype within the context of its environment. Adaptations to one environment can decrease fitness in another, revealing evolutionary trade-offs. Here, we show that the cost of gene expression underlies a trade-off between growth rate and mating efficiency in the yeast Saccharomyces cerevisiae. During asexual growth, mutations that eliminate the ability to mate provide an approximately 2% per-generation growth-rate advantage. Some strains, including most laboratory strains, carry an allele of GPA1 (an upstream component of the mating pathway) that increases mating efficiency by approximately 30% per round of mating at the cost of an approximately 1% per generation growth-rate disadvantage. In addition to demonstrating a trade-off between growth rate and mating efficiency, our results illustrate differences in the selective pressures defining fitness in the laboratory versus the natural environment and show that selection, acting on the cost of gene expression, can optimize expression levels and promote gene loss. PMID- 19299503 TI - Metal-binding loop length and not sequence dictates structure. AB - The C-terminal copper-binding loop in the beta-barrel fold of the cupredoxin azurin has been replaced with a range of sequences containing alanine, glycine, and valine residues to assess the importance of amino acid composition and the length of this region. The introduction of 2 and 4 alanines between the coordinating Cys, His, and Met results in loop structures matching those in naturally occurring proteins with the same loop lengths. A loop with 4 alanines between the Cys and His and 3 between the His and Met ligands has a structure identical to that of the WT protein, whose loop is the same length. Loop structure is dictated by length and not sequence allowing the properties of the main surface patch for interactions with partners, to which the loop is a major contributor, to be optimized. Loops with 2 amino acids between the ligands using glycine, alanine, and valine residues have been compared. An empirical relationship is found between copper site protection by the loop and reduction potential. A loop adorned with 4 methyl groups is sufficient to protect the copper ion, enabling most sequences to adequately perform this task. The mutant with 3 alanine residues between the ligands forms a strand-swapped dimer in the crystal structure, an arrangement that has not, to our knowledge, been seen previously for this family of proteins. Cupredoxins function as redox shuttles and are required to be monomeric; therefore, none have evolved with a metal binding loop of this length. PMID- 19299504 TI - Extremely low-frequency electromagnetic fields disrupt magnetic alignment of ruminants. AB - Resting and grazing cattle and deer tend to align their body axes in the geomagnetic North-South direction. The mechanism(s) that underlie this behavior remain unknown. Here, we show that extremely low-frequency magnetic fields (ELFMFs) generated by high-voltage power lines disrupt alignment of the bodies of these animals with the geomagnetic field. Body orientation of cattle and roe deer was random on pastures under or near power lines. Moreover, cattle exposed to various magnetic fields directly beneath or in the vicinity of power lines trending in various magnetic directions exhibited distinct patterns of alignment. The disturbing effect of the ELFMFs on body alignment diminished with the distance from conductors. These findings constitute evidence for magnetic sensation in large mammals as well as evidence of an overt behavioral reaction to weak ELFMFs in vertebrates. The demonstrated reaction to weak ELFMFs implies effects at the cellular and molecular levels. PMID- 19299505 TI - Structure of granzyme C reveals an unusual mechanism of protease autoinhibition. AB - Proteases act in important homeostatic pathways and are tightly regulated. Here, we report an unusual structural mechanism of regulation observed by the 2.5-A X ray crystal structure of the serine protease, granzyme C. Although the active site triad residues adopt canonical conformations, the oxyanion hole is improperly formed, and access to the primary specificity (S1) pocket is blocked through a reversible rearrangement involving Phe-191. Specifically, a register shift in the 190-strand preceding the active-site serine leads to Phe-191 filling the S1 pocket. Mutation of a unique Glu-Glu motif at positions 192-193 unlocks the enzyme, which displays chymase activity, and proteomic analysis confirms that activity of the wild-type protease can be released through interactions with an appropriate substrate. The 2.5-A structure of the unlocked enzyme reveals unprecedented flexibility in the 190-strand preceding the active-site serine that results in Phe-191 vacating the S1 pocket. Overall, these observations describe a broadly applicable mechanism of protease regulation that cannot be predicted by template-based modeling or bioinformatic approaches alone. PMID- 19299506 TI - Solution-processed, high-performance n-channel organic microwire transistors. AB - The development of solution-processable, high-performance n-channel organic semiconductors is crucial to realizing low-cost, all-organic complementary circuits. Single-crystalline organic semiconductor nano/microwires (NWs/MWs) have great potential as active materials in solution-formed high-performance transistors. However, the technology to integrate these elements into functional networks with controlled alignment and density lags far behind their inorganic counterparts. Here, we report a solution-processing approach to achieve high performance air-stable n-channel organic transistors (the field-effect mobility (mu) up to 0.24 cm(2)/Vs for MW networks) comprising high mobility, solution synthesized single-crystalline organic semiconducting MWs (mu as high as 1.4 cm(2)/Vs for individual MWs) and a filtration-and-transfer (FAT) alignment method. The FAT method enables facile control over both alignment and density of MWs. Our approach presents a route toward solution-processed, high-performance organic transistors and could be used for directed assembly of various functional organic and inorganic NWs/MWs. PMID- 19299507 TI - Simultaneous Bayesian gene tree reconstruction and reconciliation analysis. AB - We present GSR, a probabilistic model integrating gene duplication, sequence evolution, and a relaxed molecular clock for substitution rates, that enables genomewide analysis of gene families. The gene duplication and loss process is a major cause for incongruence between gene and species tree, and deterministic methods have been developed to explain such differences through tree reconciliations. Although probabilistic methods for phylogenetic inference have been around for decades, probabilistic reconciliation methods are far less established. Based on our model, we have implemented a Bayesian analysis tool, PrIME-GSR, for gene tree inference that takes a known species tree into account. Our implementation is sound and we demonstrate its utility for genomewide gene family analysis by applying it to recently presented yeast data. We validate PrIME-GSR by comparing with previous analyses of these data that take advantage of gene order information. In a case study we apply our method to the ADH gene family and are able to draw biologically relevant conclusions concerning gene duplications creating key yeast phenotypes. On a higher level this shows the biological relevance of our method. The obtained results demonstrate the value of a relaxed molecular clock. Our good performance will extend to species where gene order conservation is insufficient. PMID- 19299508 TI - Surplus value of hip adduction in leg-press exercise in patients with patellofemoral pain syndrome: a randomized controlled trial. AB - BACKGROUND: A common treatment for patients with patellofemoral pain syndrome (PFPS) is strength (force-generating capacity) training of the vastus medialis oblique muscle (VMO). Hip adduction in conjunction with knee extension is commonly used in clinical practice; however, evidence supporting the efficacy of this exercise is lacking. OBJECTIVE: The objective of this study was to determine the surplus effect of hip adduction on the VMO. DESIGN: This study was a randomized controlled trial. SETTING: The study was conducted in a kinesiology laboratory. PARTICIPANTS: Eighty-nine patients with PFPS participated. INTERVENTION: Participants were randomly assigned to 1 of 3 groups: hip adduction combined with leg-press exercise (LPHA group), leg-press exercise only (LP group), or no exercise (control group). Training consisted of 3 weekly sessions for 8 weeks. MEASUREMENTS: Ratings of worst pain as measured with a 100-mm visual analog scale (VAS-W), Lysholm scale scores, and measurements of VMO morphology (including cross-sectional area [CSA] and volume) were obtained before and after the intervention. RESULTS: Significant improvements in VAS-W ratings, Lysholm scale scores, and VMO CSA and volume were observed after the intervention in both exercise groups, but not in the control group. Significantly greater improvements for VAS-W ratings, Lysholm scale scores, and VMO volume were apparent in the LP group compared with the control group. There were no differences between the LP and LPHA groups for any measures. LIMITATIONS: Only the VMO was examined by ultrasonography. The resistance level for hip adduction and the length of intervention period may have been inadequate to induce a training effect. CONCLUSIONS: Similar changes in pain reduction, functional improvement, and VMO hypertrophy were observed in both exercise groups. Incorporating hip adduction with leg-press exercise had no impact on outcome in patients with PFPS. PMID- 19299509 TI - Life and death of sensory hair cells expressing constitutively active TRPML3. AB - The varitint-waddler mutation A419P renders TRPML3 constitutively active, resulting in cationic overload, particularly in sustained influx of Ca(2+). TRPML3 is expressed by inner ear sensory hair cells, and we were intrigued by the fact that hair cells are able to cope with expressing the TRPML3(A419P) isoform for weeks before they ultimately die. We hypothesized that the survival of varitint-waddler hair cells is linked to their ability to deal with Ca(2+) loads due to the abundance of plasma membrane calcium ATPases (PMCAs). Here, we show that PMCA2 significantly reduced [Ca(2+)](i) increase and apoptosis in HEK293 cells expressing TRPML3(A419P). The deaf-waddler isoform of PMCA2, operating at 30% efficacy, showed a significantly decreased ability to rescue the Ca(2+) loading of cells expressing TRPML3(A419P). When we combined mice heterozygous for the varitint-waddler mutant allele with mice heterozygous for the deaf-waddler mutant allele, we found severe hair bundle defects as well as increased hair cell loss compared with mice heterozygous for each mutant allele alone. Furthermore, 3 week-old double mutant mice lacked auditory brainstem responses, which were present in their respective littermates containing single mutant alleles. Likewise, heterozygous double mutant mice exhibited severe circling behavior, which was not observed in mice heterozygous for TRPML3(A419P) or PMCA2(G283S) alone. Our results provide a molecular rationale for the delayed hair cell loss in varitint-waddler mice. They also show that hair cells are able to survive for weeks with sustained Ca(2+) loading, which implies that Ca(2+) loading is an unlikely primary cause of hair cell death in ototoxic stress situations. PMID- 19299510 TI - Modifications of superoxide dismutase (SOD1) in human erythrocytes: a possible role in amyotrophic lateral sclerosis. AB - Over 100 mutations in Cu/Zn-superoxide dismutase (SOD1) result in familial amyotrophic lateral sclerosis. Dimer dissociation is the first step in SOD1 aggregation, and studies suggest nearly every amino acid residue in SOD1 is dynamically connected to the dimer interface. Post-translational modifications of SOD1 residues might be expected to have similar effects to mutations, but few modifications have been identified. Here we show, using SOD1 isolated from human erythrocytes, that human SOD1 is phosphorylated at threonine 2 and glutathionylated at cysteine 111. A second SOD1 phosphorylation was observed and mapped to either Thr-58 or Ser-59. Cysteine 111 glutathionylation promotes SOD1 monomer formation, a necessary initiating step in SOD1 aggregation, by causing a 2-fold increase in the K(d). This change in the dimer stability is expected to result in a 67% increase in monomer concentration, 315 nm rather than 212 nm at physiological SOD1 concentrations. Because protein glutathionylation is associated with redox regulation, our finding that glutathionylation promotes SOD1 monomer formation supports a model in which increased oxidative stress promotes SOD1 aggregation. PMID- 19299511 TI - Specific activation of mTORC1 by Rheb G-protein in vitro involves enhanced recruitment of its substrate protein. AB - Rheb G-protein plays critical roles in the TSC/Rheb/mTOR signaling pathway by activating mTORC1. The activation of mTORC1 by Rheb can be faithfully reproduced in vitro by using mTORC1 immunoprecipitated by the use of anti-raptor antibody from mammalian cells starved for nutrients. The low in vitro kinase activity against 4E-BP1 of this mTORC1 preparation is dramatically increased by the addition of recombinant Rheb. On the other hand, the addition of Rheb does not activate mTORC2 immunoprecipitated from mammalian cells by the use of anti-rictor antibody. The activation of mTORC1 is specific to Rheb, because other G-proteins such as KRas, RalA/B, and Cdc42 did not activate mTORC1. Both Rheb1 and Rheb2 activate mTORC1. In addition, the activation is dependent on the presence of bound GTP. We also find that the effector domain of Rheb is required for the mTORC1 activation. FKBP38, a recently proposed mediator of Rheb action, appears not to be involved in the Rheb-dependent activation of mTORC1 in vitro, because the preparation of mTORC1 that is devoid of FKBP38 is still activated by Rheb. The addition of Rheb results in a significant increase of binding of the substrate protein 4E-BP1 to mTORC1. PRAS40, a TOR signaling (TOS) motif containing protein that competes with the binding of 4EBP1 to mTORC1, inhibits Rheb-induced activation of mTORC1. A preparation of mTORC1 that is devoid of raptor is not activated by Rheb. Rheb does not induce autophosphorylation of mTOR. These results suggest that Rheb induces alteration in the binding of 4E-BP1 with mTORC1 to regulate mTORC1 activation. PMID- 19299512 TI - Bidirectional signaling through ephrinA2-EphA2 enhances osteoclastogenesis and suppresses osteoblastogenesis. AB - Bone is remodeled constantly throughout life by bone-resorbing osteoclasts and bone-forming osteoblasts. To maintain bone volume and quality, differentiation of osteoclasts and osteoblasts is tightly regulated through communication between and within these two cell lineages. Previously we reported that cell-cell interaction mediated by ephrinB2 ligand on osteoclasts and EphB4 receptor on osteoblasts generates bidirectional anti-osteoclastogenic and pro osteoblastogenic signals into respective cells and presumably facilitates transition from bone resorption to bone formation. Here we show that bidirectional ephrinA2-EphA2 signaling regulates bone remodeling at the initiation phase. EphrinA2 expression was rapidly induced by receptor activator of NF-kappaB ligand in osteoclast precursors; this was dependent on the transcription factor c-Fos but independent of the c-Fos target gene product NFATc1. Receptor EphA2 was expressed in osteoclast precursors and osteoblasts. Overexpression experiments revealed that both ephrinA2 and EphA2 in osteoclast precursors enhanced differentiation of multinucleated osteoclasts and that phospholipase Cgamma2 may mediate ephrinA2 reverse signaling. Moreover, ephrinA2 on osteoclasts was cleaved by metalloproteinases, and ephrinA2 released in the culture medium enhanced osteoclastogenesis. Interestingly, differentiation of osteoblasts lacking EphA2 was enhanced along with alkaline phosphatase, Runx2, and Osterix expression, indicating that EphA2 on osteoblasts generates anti osteoblastogenic signals presumably by up-regulating RhoA activity. Therefore, ephrinA2-EphA2 interaction facilitates the initiation phase of bone remodeling by enhancing osteoclast differentiation and suppressing osteoblast differentiation. PMID- 19299513 TI - Trolox prevents osteoclastogenesis by suppressing RANKL expression and signaling. AB - Excessive receptor activator of NF-kappaB ligand (RANKL) signaling causes enhanced osteoclast formation and bone resorption. Thus, down-regulation of RANKL expression or its downstream signals may be a therapeutic approach to the treatment of pathological bone loss. In this study, we investigated the effects of Trolox, a water-soluble vitamin E analogue, on osteoclastogenesis and RANKL signaling. Trolox potently inhibited interleukin-1-induced osteoclast formation in bone marrow cell-osteoblast coculture by abrogating RANKL induction in osteoblasts. This RANKL reduction was attributed to the reduced production of prostaglandin E(2) via a down-regulation of cyclooxygenase-2 activity. We also found that Trolox inhibited osteoclast formation from bone marrow macrophages induced by macrophage colony-stimulating factor plus RANKL in a reversible manner. Trolox was effective only when present during the early stage of culture, which implies that it targets early osteoclast precursors. Pretreatment with Trolox did not affect RANKL-induced early signaling pathways, including MAPKs, NF kappaB, and Akt. We found that Trolox down-regulated the induction by RANKL of c Fos protein by suppressing its translation. Ectopic overexpression of c-Fos rescued the inhibition of osteoclastogenesis by Trolox in bone marrow macrophages. Trolox also suppressed interleukin-1-induced osteoclast formation and bone loss in mouse calvarial bone. Taken together, our findings indicate that Trolox prevents osteoclast formation and bone loss by inhibiting both RANKL induction in osteoblasts and c-Fos expression in osteoclast precursors. PMID- 19299514 TI - Regulation of RNA polymerase III transcription involves SCH9-dependent and SCH9 independent branches of the target of rapamycin (TOR) pathway. AB - Maf1 is a conserved repressor of transcription that functions at the downstream end of multiple nutrient and stress signaling pathways. How these different signaling pathways converge on Maf1 is not known. Previous work in yeast indicates that protein kinase A (PKA) regulates RNA polymerase (pol) III transcription, in part, by phosphorylating multiple sites in Maf1. Here we present additional evidence for this view and show that a parallel nutrient and stress-sensing pathway involving Sch9, an homologous kinase to metazoan S6 kinase, targets Maf1 at a subset of PKA sites. Using ATP analog-sensitive alleles of PKA and Sch9, we find that these two kinases account for the bulk of the phosphorylation on consensus PKA sites in Maf1. Deletion of Sch9 reduces RNA pol III transcription in a Maf1-dependent manner, yet the cells remain susceptible to further repression by rapamycin and other treatments. Because the rapamycin sensitive kinase activity of the TORC1 complex is necessary for Sch9 function in vivo and in vitro, our results show that transcriptional regulation of RNA pol III and the coordinate control of ribosomal protein genes can be achieved by Sch9 dependent and -independent branches of the target of rapamycin (TOR) signaling pathway. PMID- 19299515 TI - Three homologous ArfGAPs participate in coat protein I-mediated transport. AB - ArfGAP1 is a prototype of GTPase-activating proteins for ADP-ribosylation factors (ARFs) and has been proposed to be involved in retrograde transport from the Golgi apparatus to the endoplasmic reticulum (ER) by regulating the uncoating of coat protein I (COPI)-coated vesicles. Depletion of ArfGAP1 by RNA interference, however, causes neither a discernible phenotypic change in the COPI localization nor a change in the Golgi-to-ER retrograde transport. Therefore, we also examined ArfGAP2 and ArfGAP3, closely related homologues of ArfGAP1. Cells in which ArfGAP1, ArfGAP2, and ArfGAP3 are simultaneously knocked down show an increase in the GTP-bound ARF level. Furthermore, in these cells proteins resident in or cycling through the cis-Golgi, including ERGIC-53, beta-COP, and GM130, accumulate in the ER-Golgi intermediate compartment, and Golgi-to-ER retrograde transport is blocked. The phenotypes observed in the triple ArfGAP knockdown cells are similar to those seen in beta-COP-depleted cells. Both the triple ArfGAP- and beta-COP-depleted cells accumulate characteristic vacuolar structures that are visible under electron microscope. Furthermore, COPI is concentrated at rims of the vacuolar structures in the ArfGAP-depleted cells. On the basis of these observations, we conclude that ArfGAP1, ArfGAP2, and ArfGAP3 have overlapping roles in regulating COPI function in Golgi-to-ER retrograde transport. PMID- 19299516 TI - Subunit interactions and requirements for inhibition of the yeast V1-ATPase. AB - Disassembly of the yeast V-ATPase into cytosolic V(1) and membrane V(0) sectors inactivates MgATPase activity of the V(1)-ATPase. This inactivation requires the V(1) H subunit (Parra, K. J., Keenan, K. L., and Kane, P. M. (2000) J. Biol. Chem. 275, 21761-21767), but its mechanism is not fully understood. The H subunit has two domains. Interactions of each domain with V(1) and V(0) subunits were identified by two-hybrid assay. The B subunit of the V(1) catalytic headgroup interacted with the H subunit N-terminal domain (H-NT), and the C-terminal domain (H-CT) interacted with V(1) subunits B, E (peripheral stalk), and D (central stalk), and the cytosolic N-terminal domain of V(0) subunit Vph1p. V(1)-ATPase complexes from yeast expressing H-NT are partially inhibited, exhibiting 26% the MgATPase activity of complexes with no H subunit. The H-CT domain does not copurify with V(1) when expressed in yeast, but the bacterially expressed and purified H-CT domain inhibits MgATPase activity in V(1) lacking H almost as well as the full-length H subunit. Binding of full-length H subunit to V(1) was more stable than binding of either H-NT or H-CT, suggesting that both domains contribute to binding and inhibition. Intact H and H-CT can bind to the expressed N-terminal domain of Vph1p, but this fragment of Vph1p does not bind to V(1) complexes containing subunit H. We propose that upon disassembly, the H subunit undergoes a conformational change that inhibits V(1)-ATPase activity and precludes V(0) interactions. PMID- 19299519 TI - Review of family-centered interventions to enhance the health outcomes of children with type 1 diabetes. AB - PURPOSE: The purpose of this systematic literature review was to examine family centered interventions that enhance the health outcomes of children with type 1 diabetes. The review summarizes the interventions and outcome measures that consisted of A1Cs, family relationships, and family conflict. METHODS: Multiple electronic databases were searched from their start date to September 2008 for reports of studies that examined family-centered interventions among children with type 1 diabetes. RESULTS: Nine publications, all randomized controlled trials, were located that investigated the effectiveness of family-centered interventions. Findings indicated that family-centered interventions significantly improved A1Cs, enhanced family dynamics, and decreased family conflict. CONCLUSIONS: Family-centered interventions targeting children diagnosed with type 1 diabetes appear to be effective in enhancing health outcomes. These interventions focused on traditional 2-parent families, and many did not report race or ethnicity. Given the increasing number of single-parent households and divorced parents, a need exists to explore and develop family-centered interventions that target nontraditional family structures as well as addressing cultural differences. PMID- 19299520 TI - Plasmodium falciparum and dihydrofolate reductase I164L mutations in Africa. PMID- 19299521 TI - Both a protective and a deleterious role for the L76V mutation. PMID- 19299522 TI - Development of a high-throughput brain slice method for studying drug distribution in the central nervous system. AB - New, more efficient methods of estimating unbound drug concentrations in the central nervous system (CNS) combine the amount of drug in whole brain tissue samples measured by conventional methods with in vitro estimates of the unbound brain volume of distribution (V(u,brain)). Although the brain slice method is the most reliable in vitro method for measuring V(u,brain), it has not previously been adapted for the needs of drug discovery research. The aim of this study was to increase the throughput and optimize the experimental conditions of this method. Equilibrium of drug between the buffer and the brain slice within the 4 to 5 h of incubation is a fundamental requirement. However, it is difficult to meet this requirement for many of the extensively binding, lipophilic compounds in drug discovery programs. In this study, the dimensions of the incubation vessel and mode of stirring influenced the equilibration time, as did the amount of brain tissue per unit of buffer volume. The use of cassette experiments for investigating V(u,brain) in a linear drug concentration range increased the throughput of the method. The V(u,brain) for the model compounds ranged from 4 to 3000 ml . g brain(-1), and the sources of variability are discussed. The optimized setup of the brain slice method allows precise, robust estimation of V(u,brain) for drugs with diverse properties, including highly lipophilic compounds. This is a critical step forward for the implementation of relevant measurements of CNS exposure in the drug discovery setting. PMID- 19299523 TI - The role of human hepatic cytochrome P450 isozymes in the metabolism of racemic 3,4-methylenedioxyethylamphetamine and its single enantiomers. AB - The 3,4-methylenedioxy-methamphetamine (MDMA)-related designer drug 3,4 methylenedioxyethylamphetamine (MDEA, Eve) is a chiral compound that is mainly metabolized by N-deethylation and demethylenation during phase I metabolism. The involvement of several cytochrome P450 (P450) isozymes in these metabolic steps has been demonstrated by inhibition assays using human liver microsomes. However, a comprehensive study on the involvement of all relevant human P450s has not been published yet. In addition, the chirality of this drug was not considered in these in vitro studies. The aim of the present work was first to elucidate the contribution of the relevant human P450 isozymes in the demethylenation as well as in the N-dealkylation of racemic MDEA and its single enantiomers and secondly to compare these findings with recently published data concerning the enantioselective metabolism of MDMA. Racemic MDEA and its single enantiomers were incubated using heterologously expressed human P450s, and the corresponding metabolites dihydroxyethylamphetamine and methylenedioxyamphetamine were determined by gas chromatography-mass spectrometry after chiral derivatization with S-heptafluorobutyrylprolyl chloride. The highest contributions to both metabolic steps as calculated from the enzyme kinetic data were obtained for CYP3A4 and CYP2D6 at substrate concentrations corresponding to plasma concentrations of recreational users after intake of racemic MDEA. Both metabolic reactions were found to be enantioselective with a general preference for the S enantiomers, which was particularly pronounced in the case of CYP2C19. In conclusion, different pharmacokinetic properties of MDEA enantiomers observed in vivo are therefore partially caused by P450-dependent enantioselective metabolism. PMID- 19299524 TI - Oxidized metabolites of oltipraz exert cytoprotective effects against arachidonic acid through AMP-activated protein kinase-dependent cellular antioxidant effect and mitochondrial protection. AB - Oltipraz protects cells from chemical-induced carcinogenesis partly because of phase 2 enzyme induction. Certain oltipraz metabolites also induce phase 2 enzymes. This study investigated the cytoprotective effects of the oxidized metabolites of oltipraz against arachidonic acid (AA), a proinflammatory fatty acid that causes cellular reactive oxygen species (ROS) production and mitochondrial impairment, and the mechanistic basis of their action in HepG2 cells. Treatment with 4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiol-3-one (M1) or 7 methyl-6,8-bis(methylthio)H-pyrrolo[1,2-a]-pyrazine (M2), but not 7-methyl-8 (methylsulfinyl)-6-(methylthio)H-pyrrolo[1,2-a]pyrazine (M3) or 7-methyl-6,8 bis(methylsulfinyl)H-pyrrolo[1,2-a]pyrazine (M4), enabled cells to protect against AA-induced apoptosis. M1 and M2 treatment protected cells from ROS produced by AA and inhibited AA-induced glutathione depletion. Moreover, both M1 and M2 effectively inhibited mitochondrial dysfunction induced by AA, although M2 alone slightly elicited it at a relatively high concentration. M1 and M2 activated AMP-activated protein kinase (AMPK), but M3 and M4 failed to do so. AMPK activation by M1 and M2 contributed to cell survival against AA through a decrease in cellular ROS production and prevention of mitochondrial dysfunction, as shown by the reversal of the metabolites' restoration of mitochondrial membrane potential by compound C treatment or overexpression of a dominant negative mutant AMPK. Consistently, 5-aminoimidazole-4-carboxamide-1-beta-d ribofuranoside, an AMPK activator, also had a cytoprotective and antioxidant effect against AA. Our results demonstrate that, of the major metabolites of oltipraz, M1 and M2 are capable of protecting cells from AA-induced ROS production and mitochondrial dysfunction, which may be associated with AMPK activation. PMID- 19299525 TI - Regulation of expression and activity of rat intestinal multidrug resistance associated protein 2 by cholestatic estrogens. AB - The effect of the cholestatic estrogens ethynylestradiol (EE) and estradiol 17beta-D-glucuronide (E2-17G) on expression and activity of intestinal multidrug resistant-associated protein 2 (Mrp2, Abcc2) was studied in rats. Expression and localization of Mrp2 were evaluated by Western blotting, real-time polymerase chain reaction, and confocal immunofluorescence microscopy. Mrp2 transport activity toward dinitrophenyl-S-glutathione (DNP-SG) was assessed in vitro in intestinal sacs. EE, administered subcutaneously at a 5 mg/kg b.wt. dose, for 5 consecutive days, produced a marked decrease in Mrp2 expression at post transcriptional level, without affecting its normal localization at the apical membrane of the enterocyte. This effect was selective because expression of other ATP-binding cassette proteins such as breast cancer resistance protein and Mrp3 were not affected and that of multidrug resistance protein 1 was only minimally impaired. Consistent with down-regulation of expression of Mrp2, a significant impairment in serosal to mucosal transport of DNP-SG and in protection against absorption of this same compound were registered. Simultaneous administration of EE with spironolactone (200 micromol/kg b.wt./day for 3 days), an Mrp2 inducer, prevented these alterations, confirming down-regulation of expression of Mrp2 by EE as a major component of functional changes. Incorporation of E2-17G (30 microM) in the serosal medium of intestinal sacs decreased serosal to mucosal transport of DNP-SG, probably because of competitive inhibition, without affecting normal Mrp2 expression or localization. Our data indicate impairment of function of intestinal Mrp2 by both cholestatic estrogens, although through a different mechanism. This finding represents an aggravation of deteriorated hepatic Mrp2 function that could further increase bioavailability of specific xenobiotics after oral exposure. PMID- 19299526 TI - In vitro and in silico identification and characterization of thiabendazole as a mechanism-based inhibitor of CYP1A2 and simulation of possible pharmacokinetic drug-drug interactions. AB - Thiabendazole (TBZ) and its major metabolite 5-hydroxythiabendazole (5OH-TBZ) were screened for potential time-dependent inhibition (TDI) against CYP1A2. Screen assays were carried out in the absence and presence of NADPH. TDI was observed with both compounds, with k(inact) and K(I) values of 0.08 and 0.02 min( 1) and 1.4 and 63.3 microM for TBZ and 5OH-TBZ, respectively. Enzyme inactivation was time-, concentration-, and NADPH-dependent. Inactivation by TBZ was irreversible by dialysis and oxidation by potassium ferricyanide, and there was no protection by glutathione. 5OH-TBZ was a weak TDI of CYP1A2, and enzyme activity was recovered by dialysis. IC(50) determination of TBZ and 5OH-TBZ showed both compounds to be potent inhibitors, with IC(50) values of 0.83 and 13.05 microM, respectively. IC(50) shift studies also demonstrated that TBZ was a TDI of CYP1A2. In silico methods identified the thiazole group as a TDI fragment and predicted it as the site of metabolism. The observation pointed to epoxidation of the thiazole and the benzyl rings of TBZ as possible routes of metabolism and mechanisms of TDI. Drug-drug interaction (DDI) simulation studies using SimCyp showed good predictions for competitive inhibition. However, predictions for mechanism-based inhibition (MBI)-based DDI were not in agreement with clinical observations. There was no TBZ accumulation upon chronic administration of the drug. The in vitro MBI findings might therefore not be capturing the in vivo situation in which the proposed bioactivation route is minor. This might be the case for TBZ in which, in vivo, UDP glucuronosyltransferases and sulfanotransferase metabolize and eliminate the 5OH TBZ. PMID- 19299527 TI - Ligand diversity of human and chimpanzee CYP3A4: activation of human CYP3A4 by lithocholic acid results from positive selection. AB - For currently unknown reasons, the evolution of CYP3A4 underwent acceleration in the human lineage after the split from chimpanzee. We investigated the significance of this event by comparing Escherichia coli-expressed CYP3A4 from humans, chimpanzee, and their most recent common ancestor. The expression level of chimpanzee CYP3A4 was approximately 50% of the human CYP3A4, whereas ancestral CYP3A4 did not express in E. coli. Steady-state kinetic analysis with 7 benzyloxyquinoline, 7-benzyloxy-4-(trifluoromethyl)coumarin (7-BFC), and testosterone showed no significant differences between human and chimpanzee CYP3A4. Upon addition of alpha-naphthoflavone (25 microM), human CYP3A4 showed a slightly decreased substrate concentration at which 50% of the maximal rate V(max) is reached for 7-BFC, whereas chimpanzee CYP3A4 showed a >2-fold increase. No significant differences in inhibition/activation were found for a panel of 43 drugs and endogenous compounds, suggesting that the wide substrate spectrum of human CYP3A4 precedes the human-chimpanzee split. A striking exception was the hepatotoxic secondary bile acid lithocholic acid, which at saturation caused a 5 fold increase in 7-BFC debenzylation by human CYP3A4 but not by chimpanzee CYP3A4. Mutagenesis of human CYP3A4 revealed that at least four of the six amino acids positively selected in the human lineage contribute to the activating effect of lithocholic acid. In summary, the wide functional conservation between chimpanzee and human CYP3A4 raises the prospect that phylogenetically more distant primate species such as rhesus and squirrel monkey represent suitable models of the human counterpart. Positive selection on the human CYP3A4 may have been triggered by an increased load of dietary steroids, which led to a novel defense mechanism against cholestasis. PMID- 19299528 TI - Drug offenders' perceptions of motivation: the role of motivation in rehabilitation and reintegration. AB - This article presents a qualitative analysis of participants' perceptions of the importance of motivation in the detoxification and rehabilitation process. As part of an outcome evaluation of recovering drug addicts who participated in a prison-based therapeutic community, 39 participants (a nonrandomized subsample) are interviewed regarding their rehabilitation and reintegration experiences. Although many studies show that participation in prison-based drug treatment programs reduces the likelihood of recidivism, clients in this study suggest that other factors might be equally important, and in particular clients' own motivation to change their lives. This study raises some questions about the true ability of drug treatment programs to treat and rehabilitate drug-abusing offenders, diverting the emphasis from the treatment program itself to the participants' motivation to change. Findings are discussed in regard to prison based drug treatment programs, after-release impediments encountered by inmates, and inmates' expectations of successful reintegration into the normative noncriminal society. PMID- 19299529 TI - The effects of therapeutic touch on pain. AB - PURPOSE: To better understand how Therapeutic Touch can be used in today's health care arena, this integrative literature review will examine current research that will help answer the question, Does Therapeutic Touch reduce pain? METHOD: An extensive search was conducted of the online databases MEDLINE, CINAHL, Cochrane Library, EMBASE, PsychLIT, and PubMed to retrieve research articles published from 1997 to 2007. FINDINGS: Seven studies that were conducted between 1997 and 2004 were found and only five of the seven were included as pertinent evidence to answer the question. All of the research that was reviewed to answer whether Therapeutic Touch could significantly reduce pain revealed a majority of statistically significant positive results for implementing this intervention. CONCLUSION: Because there are no identified risks to Therapeutic Touch as a pain relief measure, it is safe to recommend despite the limitations of current research. IMPLICATIONS: Therapeutic Touch should be considered among the many possible nursing interventions for the treatment of pain. PMID- 19299530 TI - The effects of feedback with and without strength training on lower extremity biomechanics. AB - BACKGROUND: Feedback instruction is a proven modality for the alteration of motion patterns. There are no existing data on the contribution of strength training, when combined with feedback instruction, to the altering of lower extremity biomechanics. HYPOTHESIS: Lower extremity muscle strength training provides an increased capacity to alter knee and hip biomechanics during a stop jump task in response to a feedback protocol. STUDY DESIGN: Controlled laboratory study. METHODS: Knee and hip 3-dimensional kinematic and kinetic data were collected for 58 female recreational athletes while performing 3 stop-jump tasks after completing a 9-week strength training program (ST-FB; n = 29) or a 9-week period of no strength training (FB; n = 29). Data were then collected for both groups after completing a jump-landing feedback instruction protocol. Knee and hip joint angles, as well as resultant forces and moments, were calculated. RESULTS: Across all participants, there were decreased peak vertical ground reaction forces (P < .001) and increased knee flexion (P = .050), hip flexion (P < .001), and hip abduction (P = .032) angles, subsequent to the feedback protocol. Hip abduction angle (P < .001) increased in the ST-FB group but not the FB group, and peak knee anterior shear force (P = .015) decreased in the ST-FB group but increased in the FB group (P = .009). CONCLUSION: The results indicate that strength training, when used in conjunction with video-assisted feedback, may provide an increased capacity for the alteration of knee and hip biomechanics. CLINICAL RELEVANCE: Programs that include both strength training and movement education through feedback may be necessary to increase the effectiveness of anterior cruciate ligament prevention programs. Strength training may provide an increased capacity for athletes to respond to other intervention modalities used in anterior cruciate ligament injury prevention programs. PMID- 19299531 TI - Traumatic and overuse injuries among international elite junior rowers. AB - BACKGROUND: Junior rowers have competed internationally for over 4 decades, and there are no epidemiological data available on traumatic and overuse injury in this population. OBJECTIVE: To define the types of musculoskeletal problems present in international elite-level junior rowers and to determine whether gender, physical stature, rowing discipline, and training programs affect the incidence of reported injuries. STUDY DESIGN: Descriptive epidemiology study. METHODS: Injury data were obtained from a total of 398 rowers (42% female, 58% male) who completed a 4-page questionnaire on injury incidence while participating at the Junior World Rowing Championships in Beijing, People's Republic of China, in August 2007. RESULTS: Overall, 290 (73.8%) reported injuries involved overuse, and 103 (26.2%) were related to a single traumatic event. Female rowers were injured more frequently than male rowers (110.2 vs 90.5 injuries per 100 rowers). In both genders, the most common injury site was the low back followed by the knee and the forearm/wrist. The severity of reported injuries was incidental in 65.1%, minor in 21.4%, moderate in 10.4%, and major in 3.1% of cases. The rowers with traumatic injuries had less rowing experience than the uninjured rowers (median [C] +/- interquartile range [Q] = 3 +/- 3 years vs 4 +/- 3 years; P = .043, Mann-Whitney test). Sweep rowers who changed rowing side during the current season had significantly more acute-onset low back injuries (P = .012, chi(2) test) than those who did not change rowing side during the same period. The incidence of traumatic injuries was significantly lower in rowers who regularly performed more than 10 minutes of posttraining stretching (P = .030, chi2) test). Athletes who ran more than once a week had more overuse knee injuries than those who ran once or less per week (P = .033, chi2 test). CONCLUSION: Elite junior rowers attending the World Rowing Championships reported predominantly overuse injuries of low severity during the current rowing season. Low back injuries were the most frequent complaint of elite-level junior rowers. PMID- 19299532 TI - Prediction of human pharmacokinetics from preclinical information: comparative accuracy of quantitative prediction approaches. AB - Quantitative prediction of human pharmacokinetics is critical in assessing the viability of drug candidates and in determining first-in-human dosing. Numerous prediction methodologies, incorporating both in vitro and preclinical in vivo data, have been developed in recent years, each with advantages and disadvantages. However, the lack of a comprehensive data set, both preclinical and clinical, has limited efforts to evaluate the optimal strategy (or strategies) that results in quantitative predictions of human pharmacokinetics. To address this issue, the authors conducted a retrospective analysis using 50 proprietary compounds for which in vitro, preclinical pharmacokinetic data and oral single-dose human pharmacokinetic data were available. Five predictive strategies, involving either allometry or use of unbound intrinsic clearance from microsomes or hepatocytes, were then compared for their ability to predict human oral clearance, half-life through predictions of systemic clearance, volume of distribution, and bioavailability. Use of a single-species scaling approach with rat, dog, or monkey was as accurate as or more accurate than using multiple species allometry. For those compounds cleared almost exclusively by P450 mediated pathways, scaling from human liver microsomes was as predictive as single-species scaling of clearance based on data from rat, dog, or monkey. These data suggest that use of predictive methods involving either single-species in vivo data or in vitro human liver microsomes can quantitatively predict human in vivo pharmacokinetics and suggest the possibility of streamlining the predictive methodology through use of a single species or use only of human in vitro microsomal preparations. PMID- 19299533 TI - Clinically important interaction between tedisamil and verapamil. AB - Tedisamil, a class III antiarrhythmic drug, is a P-glycoprotein substrate. Tedisamil treatment may implicate coadministration with class IV antiarrhythmics such as verapamil, a P-glycoprotein inhibitor. Pharmacokinetic and pharmacodynamic interactions between tedisamil and verapamil were evaluated in a double-blind, crossover study. Twelve healthy volunteers received a 3-day treatment of tedisamil (100 mg bid), verapamil (180 mg bid), a combination of these drugs, or placebo. Blood pressure and electrocardiograms were assessed daily and cardiac output and pharmacokinetics on day 3. Combination of tedisamil and verapamil increased tedisamil plasma concentrations (AUC(0-12 h): +77%, CI(90%): +51% to +108%; C(max): +78%, CI(90%): +57% to +102%) compared to tedisamil monotreatment but decreased plasma concentrations of verapamil (AUC(0 12 h): -21%, CI(90%): -32% to -8%; C(max): -28%, CI(90%): -39% to -14%) and norverapamil (AUC(0-12 h): -17%, CI(90%): -28% to -6%; C(max): -20%, CI(90%):-29% to -10%) compared to verapamil monotreatment. Compared to placebo, verapamil and the combination treatment increased PR by 23.5 (CI(95%): 17.9 to 29.2) ms and 12.2 (5.7 to 17.0) ms, respectively. Compared to placebo, tedisamil and the combination treatment increased QTc by 27.8 (15.8 to 39.8) ms and 45.7 (33.7 to 57.7) ms, respectively. Thus, concomitant use of tedisamil with P-glycoprotein inhibitors likely results in clinically significant drug interactions. PMID- 19299534 TI - Trigeminal neuralgia treated with pregabalin in family medicine settings: its effect on pain alleviation and cost reduction. AB - The purpose of this study is to analyze the effect of pregabalin (PGB) on pain alleviation, use of health care and non-health care resources, and associated costs in patients with trigeminal neuralgia under usual clinical practice in primary care settings. Sixty-five PGB-naive patients receiving PGB as monotherapy (n = 36, 55%) or combined with other drugs (n = 29, 45%) fulfill criteria for inclusion in a secondary analysis from a 12-week, multicenter, observational prospective study aimed to ascertain the cost of illness in subjects with neuropathic pain. Pain is evaluated using the Short Form McGill Pain Questionnaire. Use of health care and non-health care resources and lost workdays equivalents (LWDEs) are also recorded. PGB significantly reduces pain scores, use of health care resources (ancillary tests and unscheduled medical visits), and number of LWDEs. Additional cost of PGB treatment (+euro 174 +/- 106) is broadly compensated for by a reduction in both health care costs (-euro 621 +/-1211, P < .001) and indirect costs (-euro 1210 +/- 1141, P < .001). It is concluded that PGB as monotherapy or combined with other drugs is effective in pain management in patients with trigeminal neuralgia and reduces the cost of illness. PMID- 19299535 TI - The "hijacking" of moral conscience from pharmacy practice: a Canadian perspective. AB - The new vision for Canadian pharmacy practice, as outlined in the Blueprint for Pharmacy, drives for a more patient-centered and outcomes-focused approach to patient care than ever before. As of 2007 and 2009, pharmacists in Alberta and British Columbia, respectively, have the ability to adapt and renew existing prescriptions without prior approval from the physician. This necessarily puts a greater emphasis on the practice of pharmaceutical care, entailing the formation of a closer professional, more personalized bonding between pharmacist and patient. No one, however, seems to be paying much attention to the greater moral and ethical responsibility of pharmacists as they take on expanded roles. Not only are the greater expectations not coupled with greater respect for the pharmacist's freedom of conscience, but provincial jurisdictions across the country vary in degrees of tolerance toward pharmacists whose views differ from those held by the licensing authorities. PMID- 19299536 TI - Lowly expressed human microRNA genes evolve rapidly. AB - To study the evolution of human microRNAs (miRNAs), we examined nucleotide variation in humans, sequence divergence between species, and genomic clustering patterns for miRNAs with different expression levels. We found that expression level is a major indicator of the rate of evolution and that approximately 30% of currently annotated human miRNA genes are almost free of selective pressure. PMID- 19299537 TI - Predicting stages of change in battered women. AB - Battered women's stages of change (SOCs) are examined in this study. First, confirmatory factor analysis and latent profile analysis were conducted on 754 battered women's responses on the Problems in Relationship Scale (Brown, 1998). Factor loadings were strong, and latent variable mixture modeling produces a two class solution. Second, comparisons of women (including a subset of 276) suggest that they were relatively advanced in SOC. SOC was unrelated to intimate partner violence experiences, but an earlier stage was associated with ethnicity, economic and emotional dependence, preoccupied attachment, satisfaction with social supports, and women's use of aggression. Implications for interventions and future research are described. PMID- 19299538 TI - Assay configuration and analytic specificity may have major effects on prediction of clinical outcomes-- implications for reference standards. PMID- 19299540 TI - Grading quality of evidence and strength of recommendations for diagnostic tests and strategies. PMID- 19299541 TI - Salivary leptin as a candidate diagnostic marker in salivary gland tumors. AB - BACKGROUND: Since the discovery of autonomous leptin production in salivary glands, very few studies have reported on the physiological or pathological meaning of this particular cytokine in saliva. The aim of this study was to investigate the expression of leptin and its receptors Ob-Ra and Ob-Rb in parotid salivary gland tumors, with particular regard to a potential use of leptin as a tumor marker. METHODS: Parotid tissue samples from healthy individuals (n = 31) and tumor patients (n = 97, including tissue samples from pleomorphic adenomas, adenolymphomas, basal cell adenomas, and diverse carcinomas) were analyzed by use of ApoTome-technique microscopy, immunohistochemistry, immunofluorescence, immunoblotting, and quantitative real-time PCR. Salivary and plasma leptin concentrations were measured by using ELISA. Ultrasound was used to determine tumor size before surgery. RESULTS: In all salivary gland tumors leptin was expressed in much higher amounts than in healthy parotid tissues. The cytokine was not imported from the blood but actively produced by the tumors. Immunoblotting results indicated that leptin was present as oligomers in salivary glands. Furthermore, the examined tumors overexpressed the receptor isoforms Ob Ra and Ob-Rb. Measured leptin concentrations in mixed saliva samples were significantly increased in patients with parotid tumors [mean (SD) 673 (484) pg/mL in pleomorphic adenomas, 679 (465) pg/mL in adenolymphomas, and 880 (618) pg/mL in carcinomas] vs controls [125 (36) pg/mL] (P < 0.001). CONCLUSIONS: This is the first study to show that the analysis of salivary leptin in mixed saliva samples may allow preoperative differentiation between tumor patients and healthy individuals. PMID- 19299542 TI - Role of monitoring changes in sensitive cardiac troponin I assay results for early diagnosis of myocardial infarction and prediction of risk of adverse events. AB - BACKGROUND: We sought to determine the diagnostic accuracy of the cardiac troponin I (cTnI) VITROS(R) Troponin I-ES assay for early detection of acute myocardial infarction (AMI) and for risk prediction of adverse events in patients with symptoms of acute coronary syndrome (ACS). METHODS: cTnI was measured on admission and approximately 6 h postadmission in 381 patients. The 99th percentile cTnI concentration (0.034 microg/L) and change [delta (delta)] between admission and follow-up concentrations were evaluated in diagnostic sensitivity and specificity calculations. Risk of cardiac event or death within 60 days was evaluated by Cox proportional hazards regression. RESULTS: AMI occurred in 52 patients. Diagnostic sensitivities (95% CI) of admission and follow-up cTnIs for AMI were 69% (55%-81%) and 94% (84%-99%), respectively. The corresponding specificities (95% CI) were 78% (73%-82%) and 81% (77%-85%), and ROC curve areas were 0.82 vs 0.96 (P < 0.001). Deltas between admission and follow-up cTnI >30% had a sensitivity of 75% (95% CI 61%-86%) and a specificity of 91% (95% CI 87% 94%). During follow-up, 1 cardiac death, 2 noncardiac deaths, 52 AMIs, 6 coronary artery bypass grafts, and 43 percutanous coronary interventions occurred in 62 patients. A delta cTnI >30%, when added to either initial cTnI >0.034 microg/L or follow-up cTnI >0.034 microg/L, improved risk stratification for cardiac event or death (P < 0.001). CONCLUSIONS: Admission cTnI measured by the VITROS ES assay is a sensitive biomarker for detection of AMI. Utilizing >30% cTnI delta in addition to either the baseline or follow-up concentration improved both specificity and risk assessment in patients presenting with symptoms of ACS. PMID- 19299543 TI - A new principle for measurement of cobalamin and corrinoids, used for studies of cobalamin analogs on serum haptocorrin. AB - BACKGROUND: Transcobalamin (TC) and haptocorrin (HC) are serum corrinoid-binding proteins. We developed new methods for measurement of the corrinoids bound to HC and TC. METHODS: TC (n = 10) or HC (n = 138) was immunoprecipitated, and corrinoids were released by enzymatic degradation [subtilisin Carlsberg (EC 3.4.21.62)] of the binding proteins. Binding of the released corrinoids to added unsaturated TC (apoTC) or HC (apoHC) created holoTC (as measure of cobalamins) and holoHC (as measure of corrinoids). holoTC and holoHC were measured by use of ELISA. The amounts of analogs were calculated as the difference between corrinoids and cobalamins. Corrinoids extracted from HC were separated with HPLC after addition of potassium cyanide (n = 3). RESULTS: The corrinoid- and cobalamin-specific assays had a positive linear relation between analyte concentration and assay signal, detection limits of 8 and 4 pmol/L, and imprecision values (CV) of 25 mmHg and occurred at a rate of 15-25 h(-1) of which over 50% were aborally propagative. Responses during the first 20-min of RS were characterized by high-frequency and high-amplitude contractions that were correlated with defecation. RS-induced GMCs and fecal pellet output were blocked by atropine (0.5 mg/kg ip) or the corticotrophin releasing factor (CRF) receptor antagonist astressin-B (100 microg/kg ip). RS activated colonic myenteric neurons as shown by Fos immunoreactivity. In mice previously exposed to repeated RS (60 min/day, 14 days), or in transgenic mice that overexpress CRF, the duration of stimulation of phasic colonic contractions was significantly shorter (10 vs. 20 min). In contrast to RS, abdominal surgery abolished colonic contractions including GMCs. These findings provide the first evidence for the presence of frequent cholinergic-dependent GMCs in the distal colon of conscious mice and their modulation by acute and chronic stressors. Noninvasive colonic manometry opens new venues to investigate colonic motor function in genetically modified mice relevant to diseases that involve colonic motility alterations. PMID- 19299580 TI - Endothelin antagonism in portal hypertensive mice: implications for endothelin receptor-specific signaling in liver disease. AB - Endothelin-1 (ET-1), a potent vasoactive peptide, plays an important role in the pathogenesis of liver disease and portal hypertension. Two major endothelin receptors (ET-A and ET-B) mediate biological effects, largely on the basis of their known downstream signaling pathways. We hypothesized that the different receptors are likely to mediate divergent effects in portal hypertensive mice. Liver fibrosis and cirrhosis and portal hypertension were induced in 8-wk-old male BALB/c mice by gavage with carbon tetrachloride (CCl4). Portal pressure was recorded acutely during intravenous infusion of endothelin receptor antagonists in normal or portal hypertensive mice. In vivo microscopy was used to monitor sinusoidal dynamics. Additionally, the effect of chronic exposure to endothelin antagonists was assessed in mice during induction of fibrosis and cirrhosis with CCl4 for 8 wk. Intravenous infusion of ET-A receptor antagonists into normal and cirrhotic mice reduced portal pressure whereas ET-B receptor antagonism increased portal pressure. A mixed endothelin receptor antagonist also significantly reduced portal pressure. Additionally, the ET-A receptor antagonist caused sinusoidal dilation, whereas the ET-B receptor antagonist caused sinusoidal constriction. Chronic administration of each the endothelin receptor antagonists during the induction of fibrosis and portal hypertension led to reduced fibrosis, a significant reduction in portal pressure, and altered sinusoidal dynamics relative to controls. Acute effects of endothelin receptor antagonists are likely directly on the hepatic and sinusoidal vasculature, whereas chronic endothelin receptor antagonism appears to be more complicated, likely affecting fibrogenesis and the hepatic microcirculation. The data imply a relationship between hepatic fibrogenesis or fibrosis and vasomotor responses. PMID- 19299581 TI - Inflammation-induced, 3'UTR-dependent translational inhibition of Hsp70 mRNA impairs intestinal homeostasis. AB - Although the inducible heat shock protein 70 (Hsp70) is essential for maintaining intestinal homeostasis in colitis, it is translationally downregulated in inflamed colonic mucosa, paradoxically rendering the gut more susceptible to injury. We examined the basis for this process by analyzing the role of untranslated regions (UTR) of Hsp70 mRNA in inflammation-associated downregulation in vitro and in vivo. Using luciferase-reporter assays in young adult mouse intestinal epithelial cells, we determined that cytokine-induced translational inhibition of Hsp70 mRNA was mediated by the 3'UTR, but not 5'UTR. In vivo, dextran sodium sulfate (DSS) colitis was induced in wild-type (WT) and villin-promoter regulated "UTR-less" Hsp70 transgenic (TG) mice, the latter exhibiting intestinal epithelial-specific transgene expression. Progressive downregulation of colonic Hsp70 protein expression was observed in WT, but not in TG, mice with increasing severity of mucosal inflammation, confirming the essential role of the 3'UTR in mediating inflammation-associated downregulation of Hsp70. Hsp70 TG mice demonstrated significantly lower endoscopic and histological inflammation scores in DSS-induced colitis than WT. In conclusion, downregulation of Hsp70 expression in inflamed mucosa is mediated by translational inhibition requiring the 3'UTR, resulting in increased mucosal injury. By forcing intestinal epithelial-specific Hsp70 expression in vivo, the severity of experimentally induced colitis was significantly reduced. PMID- 19299582 TI - Role of SIRT1 in regulation of LPS- or two ethanol metabolites-induced TNF-alpha production in cultured macrophage cell lines. AB - Dysregulation of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) has been implicated in the pathogenesis of alcoholic liver injury. Sirtuin 1 (SIRT1) is an NAD(+)-dependent class III protein deacetylase that is known to be involved in regulating production of proinflammatory cytokines including TNF-alpha. In the present study, we examined the role of SIRT1 signaling in TNF-alpha generation stimulated by either lipopolysaccharide (LPS), acetaldehyde (AcH), or acetate (two major metabolites of ethanol) in two cultured macrophage cell lines. In both rat Kupffer cell line 1 (RKC1) and murine RAW 264.7 macrophages, treatment with either LPS, AcH, or acetate caused significant decreases in SIRT1 transcription, translation, and activation, which essentially demonstrated an inverse relationship with TNF-alpha levels. LPS, AcH, and acetate each provoked the release of TNF-alpha from RKC1 cells, whereas coincubation with resveratrol (a potent SIRT1 agonist) inhibited this effect. Conversely, addition of sirtinol (a known SIRT1 inhibitor) or knocking down SIRT1 by the small silencing SIRT1 plasmid (SIRT1shRNA) augmented TNF-alpha release, suggesting that impairment of SIRT1 may contribute to TNF-alpha secretion. Further mechanistic studies revealed that inhibition of SIRT1 by LPS, AcH, or acetate was associated with a marked increase in the acetylation of the RelA/p65 subunit of nuclear transcription factor (NF-kappaB) and promotion of NF-kappaB transcriptional activity. Taken together, our findings suggest that SIRT1-NF-kappaB signaling is involved in regulating LPS- and metabolites-of-ethanol-mediated TNF-alpha production in rat Kupffer cells and in murine macrophages. Our study provides new insights into understanding the molecular mechanisms underlying the development of alcoholic steatohepatitis. PMID- 19299583 TI - Circadian clock feedback cycle through NAMPT-mediated NAD+ biosynthesis. AB - The circadian clock is encoded by a transcription-translation feedback loop that synchronizes behavior and metabolism with the light-dark cycle. Here we report that both the rate-limiting enzyme in mammalian nicotinamide adenine dinucleotide (NAD+) biosynthesis, nicotinamide phosphoribosyltransferase (NAMPT), and levels of NAD+ display circadian oscillations that are regulated by the core clock machinery in mice. Inhibition of NAMPT promotes oscillation of the clock gene Per2 by releasing CLOCK:BMAL1 from suppression by SIRT1. In turn, the circadian transcription factor CLOCK binds to and up-regulates Nampt, thus completing a feedback loop involving NAMPT/NAD+ and SIRT1/CLOCK:BMAL1. PMID- 19299584 TI - Benzothiazinones kill Mycobacterium tuberculosis by blocking arabinan synthesis. AB - New drugs are required to counter the tuberculosis (TB) pandemic. Here, we describe the synthesis and characterization of 1,3-benzothiazin-4-ones (BTZs), a new class of antimycobacterial agents that kill Mycobacterium tuberculosis in vitro, ex vivo, and in mouse models of TB. Using genetics and biochemistry, we identified the enzyme decaprenylphosphoryl-beta-d-ribose 2'-epimerase as a major BTZ target. Inhibition of this enzymatic activity abolishes the formation of decaprenylphosphoryl arabinose, a key precursor that is required for the synthesis of the cell-wall arabinans, thus provoking cell lysis and bacterial death. The most advanced compound, BTZ043, is a candidate for inclusion in combination therapies for both drug-sensitive and extensively drug-resistant TB. PMID- 19299585 TI - gamma-Secretase heterogeneity in the Aph1 subunit: relevance for Alzheimer's disease. AB - The gamma-secretase complex plays a role in Alzheimer's disease and cancer progression. The development of clinically useful inhibitors, however, is complicated by the role of the gamma-secretase complex in regulated intramembrane proteolysis of Notch and other essential proteins. Different gamma-secretase complexes containing different Presenilin or Aph1 protein subunits are present in various tissues. Here we show that these complexes have heterogeneous biochemical and physiological properties. Specific inactivation of the Aph1B gamma-secretase in a mouse Alzheimer's disease model led to improvements of Alzheimer's disease relevant phenotypic features without any Notch-related side effects. The Aph1B complex contributes to total gamma-secretase activity in the human brain, and thus specific targeting of Aph1B-containing gamma-secretase complexes may help generate less toxic therapies for Alzheimer's disease. PMID- 19299586 TI - The disappearance of the progenitors of supernovae 1993J and 2003gd. AB - Using images from the Hubble Space Telescope and the Gemini Telescope, we confirmed the disappearance of the progenitors of two type II supernovae (SNe) and evaluated the presence of other stars associated with them. We found that the progenitor of SN 2003gd, an M-supergiant star, is no longer observed at the SN location and determined its intrinsic brightness using image subtraction techniques. The progenitor of SN 1993J, a K-supergiant star, is also no longer present, but its B-supergiant binary companion is still observed. The disappearance of the progenitors confirms that these two supernovae were produced by red supergiants. PMID- 19299587 TI - Optical deconstruction of parkinsonian neural circuitry. AB - Deep brain stimulation (DBS) is a therapeutic option for intractable neurological and psychiatric disorders, including Parkinson's disease and major depression. Because of the heterogeneity of brain tissues where electrodes are placed, it has been challenging to elucidate the relevant target cell types or underlying mechanisms of DBS. We used optogenetics and solid-state optics to systematically drive or inhibit an array of distinct circuit elements in freely moving parkinsonian rodents and found that therapeutic effects within the subthalamic nucleus can be accounted for by direct selective stimulation of afferent axons projecting to this region. In addition to providing insight into DBS mechanisms, these results demonstrate an optical approach for dissection of disease circuitry and define the technological toolbox needed for systematic deconstruction of disease circuits by selectively controlling individual components. PMID- 19299588 TI - The enlightenment returns. PMID- 19299589 TI - Global warming: Projections of climate change go from bad to worse, scientists report. PMID- 19299590 TI - Mathematical sciences. Senate majority leader hands NSF a gift to serve the exceptionally gifted. PMID- 19299591 TI - Public health. Stronger research just one item on drug agency's wish list. PMID- 19299592 TI - Archaeology. Recipe for rice domestication required millennia. PMID- 19299593 TI - Solar physics. Unseen link may solve the mystery of the sun's superhot corona. PMID- 19299595 TI - Obama executive order. For Congress and NIH, headaches ahead on stem cells. PMID- 19299596 TI - Australia. Stem cell center looks to recast itself in supporting role. PMID- 19299598 TI - Neuropsychiatry. Rewiring faulty circuits in the brain. PMID- 19299599 TI - Newsmaker interview. A lifetime of work gone to waste? PMID- 19299600 TI - Conservation biology. Exxon Valdez turns 20. PMID- 19299601 TI - Pandemic influenza: an inconvenient mutation. PMID- 19299602 TI - Romanian expatriates face career obstacles. PMID- 19299603 TI - Reversible exploration not worth the cost. PMID- 19299604 TI - Medicine. Monitoring and regulating offshore stem cell clinics. PMID- 19299605 TI - Materials science. Flexible electronics. PMID- 19299606 TI - Immunology. Two-in-one designer antibodies. PMID- 19299607 TI - Molecular biology. Dynamic DNA methylation. PMID- 19299608 TI - Physics. Fullerides in a squeeze. PMID- 19299609 TI - Materials science. Stiffer than steel. PMID- 19299610 TI - Chemistry. Copper puts arenes in a hard position. PMID- 19299611 TI - Flagellum mediates symbiosis. AB - We report here molecular mechanisms underlying a bacteria-archaeon symbiosis. We found that a fermentative bacterium used its flagellum for interaction with a specific methanogenic archaeon. The archaeon perceived a bacterial flagellum protein and activated its metabolism (methanogenesis). Transcriptome analyses showed that a substantial number of genes in the archaeon, including those involved in the methanogenesis pathway, were up-regulated after the contact with the flagellum protein. These findings suggest that the bacterium communicates with the archaeon by using its flagellum. PMID- 19299612 TI - Giant-stroke, superelastic carbon nanotube aerogel muscles. AB - Improved electrically powered artificial muscles are needed for generating force, moving objects, and accomplishing work. Carbon nanotube aerogel sheets are the sole component of new artificial muscles that provide giant elongations and elongation rates of 220% and (3.7 x 10(4))% per second, respectively, at operating temperatures from 80 to 1900 kelvin. These solid-state-fabricated sheets are enthalpic rubbers having gaslike density and specific strength in one direction higher than those of steel plate. Actuation decreases nanotube aerogel density and can be permanently frozen for such device applications as transparent electrodes. Poisson's ratios reach 15, a factor of 30 higher than for conventional rubbers. These giant Poisson's ratios explain the observed opposite sign of width and length actuation and result in rare properties: negative linear compressibility and stretch densification. PMID- 19299613 TI - Spinal cord stimulation restores locomotion in animal models of Parkinson's disease. AB - Dopamine replacement therapy is useful for treating motor symptoms in the early phase of Parkinson's disease, but it is less effective in the long term. Electrical deep-brain stimulation is a valuable complement to pharmacological treatment but involves a highly invasive surgical procedure. We found that epidural electrical stimulation of the dorsal columns in the spinal cord restores locomotion in both acute pharmacologically induced dopamine-depleted mice and in chronic 6-hydroxydopamine-lesioned rats. The functional recovery was paralleled by a disruption of aberrant low-frequency synchronous corticostriatal oscillations, leading to the emergence of neuronal activity patterns that resemble the state normally preceding spontaneous initiation of locomotion. We propose that dorsal column stimulation might become an efficient and less invasive alternative for treatment of Parkinson's disease in the future. PMID- 19299614 TI - Alfven waves in the lower solar atmosphere. AB - The flow of energy through the solar atmosphere and the heating of the Sun's outer regions are still not understood. Here, we report the detection of oscillatory phenomena associated with a large bright-point group that is 430,000 square kilometers in area and located near the solar disk center. Wavelet analysis reveals full-width half-maximum oscillations with periodicities ranging from 126 to 700 seconds originating above the bright point and significance levels exceeding 99%. These oscillations, 2.6 kilometers per second in amplitude, are coupled with chromospheric line-of-sight Doppler velocities with an average blue shift of 23 kilometers per second. A lack of cospatial intensity oscillations and transversal displacements rules out the presence of magneto acoustic wave modes. The oscillations are a signature of Alfven waves produced by a torsional twist of +/-22 degrees. A phase shift of 180 degrees across the diameter of the bright point suggests that these torsional Alfven oscillations are induced globally throughout the entire brightening. The energy flux associated with this wave mode is sufficient to heat the solar corona. PMID- 19299615 TI - The disorder-free non-BCS superconductor Cs3C60 emerges from an antiferromagnetic insulator parent state. AB - The body-centered cubic A15-structured cesium fulleride Cs3C60 is not superconducting at ambient pressure and is free from disorder, unlike the well studied face-centered cubic A3C60 alkali metal fulleride superconductors. We found that in Cs3C60, where the molecular valences are precisely assigned, the superconducting state at 38 kelvin emerges directly from a localized electron antiferromagnetic insulating state with the application of pressure. This transition maintains the threefold degeneracy of the active orbitals in both competing electronic states; it is thus a purely electronic transition to a superconducting state, with a dependence of the transition temperature on pressure-induced changes of anion packing density that is not explicable by Bardeen-Cooper-Schrieffer (BCS) theory. PMID- 19299616 TI - A meta-selective copper-catalyzed C-H bond arylation. AB - For over a century, chemical transformations of benzene derivatives have been guided by the high selectivity for electrophilic attack at the ortho/para positions in electron-rich substrates and at the meta position in electron deficient molecules. We have developed a copper-catalyzed arylation reaction that, in contrast, selectively substitutes phenyl electrophiles at the aromatic carbon-hydrogen sites meta to an amido substituent. This previously elusive class of transformation is applicable to a broad range of aromatic compounds. PMID- 19299617 TI - The Burgess Shale anomalocaridid Hurdia and its significance for early euarthropod evolution. AB - As the largest predators of the Cambrian seas, the anomalocaridids had an important impact in structuring the first complex marine animal communities, but many aspects of anomalocaridid morphology, diversity, ecology, and affinity remain unclear owing to a paucity of specimens. Here we describe the anomalocaridid Hurdia, based on several hundred specimens from the Burgess Shale in Canada. Hurdia possesses a general body architecture similar to those of Anomalocaris and Laggania, including the presence of exceptionally well-preserved gills, but differs from those anomalocaridids by possessing a prominent anterior carapace structure. These features amplify and clarify the diversity of known anomalocaridid morphology and provide insight into the origins of important arthropod features, such as the head shield and respiratory exites. PMID- 19299618 TI - Genetic incompatibility drives sex allocation and maternal investment in a polymorphic finch. AB - Genetic compatibility may drive individual mate choice decisions because of predictable fitness effects associated with breeding with incompatible partners. In Gouldian finches (Erythrura gouldiae), females paired with genetically incompatible males of alternative color morphs overproduce sons, presumably to reduce investment in inviable daughters. We also observed a reduced overall investment in clutch size, egg size, and care to offspring resulting from incompatible matings. Within-female experimental pairings demonstrate that female birds have the ability to adaptively adjust the sex of their eggs and allocate resources on the basis of partner quality. Female Gouldian finches thus make cumulative strategic allocation decisions to minimize the costs of poor-quality pairings when faced with a genetically incompatible partner. PMID- 19299619 TI - The domestication process and domestication rate in rice: spikelet bases from the Lower Yangtze. AB - The process of rice domestication occurred in the Lower Yangtze region of Zhejiang, China, between 6900 and 6600 years ago. Archaeobotanical evidence from the site of Tianluoshan shows that the proportion of nonshattering domesticated rice (Oryza sativa) spikelet bases increased over this period from 27% to 39%. Over the same period, rice remains increased from 8% to 24% of all plant remains, which suggests an increased consumption relative to wild gathered foods. In addition, an assemblage of annual grasses, sedges, and other herbaceous plants indicates the presence of arable weeds, typical of cultivated rice, that also increased over this period. PMID- 19299620 TI - Variants of the antibody herceptin that interact with HER2 and VEGF at the antigen binding site. AB - The interface between antibody and antigen is often depicted as a lock and key, suggesting that an antibody surface can accommodate only one antigen. Here, we describe an antibody with an antigen binding site that binds two distinct proteins with high affinity. We isolated a variant of Herceptin, a therapeutic monoclonal antibody that binds the human epidermal growth factor receptor 2 (HER2), on the basis of its ability to simultaneously interact with vascular endothelial growth factor (VEGF). Crystallographic and mutagenesis studies revealed that distinct amino acids of this antibody, called bH1, engage HER2 and VEGF energetically, but there is extensive overlap between the antibody surface areas contacting the two antigens. An affinity-improved version of bH1 inhibits both HER2- and VEGF-mediated cell proliferation in vitro and tumor progression in mouse models. Such "two-in-one" antibodies challenge the monoclonal antibody paradigm of one binding site, one antigen. They could also provide new opportunities for antibody-based therapy. PMID- 19299621 TI - Ankyrin-G promotes cyclic nucleotide-gated channel transport to rod photoreceptor sensory cilia. AB - Cyclic nucleotide-gated (CNG) channels localize exclusively to the plasma membrane of photosensitive outer segments of rod photoreceptors where they generate the electrical response to light. Here, we report the finding that targeting of CNG channels to the rod outer segment required their interaction with ankyrin-G. Ankyrin-G localized exclusively to rod outer segments, coimmunoprecipitated with the CNG channel, and bound to the C-terminal domain of the channel beta1 subunit. Ankyrin-G depletion in neonatal mouse retinas markedly reduced CNG channel expression. Transgenic expression of CNG channel beta-subunit mutants in Xenopus rods showed that ankyrin-G binding was necessary and sufficient for targeting of the beta1 subunit to outer segments. Thus, ankyrin-G is required for transport of CNG channels to the plasma membrane of rod outer segments. PMID- 19299622 TI - The surprising power of neighborly advice. AB - Two experiments revealed that (i) people can more accurately predict their affective reactions to a future event when they know how a neighbor in their social network reacted to the event than when they know about the event itself and (ii) people do not believe this. Undergraduates made more accurate predictions about their affective reactions to a 5-minute speed date (n = 25) and to a peer evaluation (n = 88) when they knew only how another undergraduate had reacted to these events than when they had information about the events themselves. Both participants and independent judges mistakenly believed that predictions based on information about the event would be more accurate than predictions based on information about how another person had reacted to it. PMID- 19299625 TI - Enhanced capture of extramembranous IgM and IgG on B cells in the NOD mouse- implications for immune complex trapping. AB - Binding of various antibody isotypes to B cells through either FcgammaRs or complement receptors has been attributed to play several roles, e.g. in immune complex (IC) transportation and regulation of B cell receptor signaling. We have revealed a novel B cell intrinsic receptor for IgM and IgG which is present in C57BL/6 (B6) mice and is more abundant in non-obese diabetic (NOD) mice. As a consequence, the level of extramembranous IgG monomers and IgM pentamers on peripheral blood B cells from NOD mice was significantly higher compared with B6 mice. The effect of this aberration was that all B cells in peripheral blood of (NOD.IgH(a) x B6(IgH(b)))F(1) mice carried both IgM allotypes on their surface. In addition, analysis of IC binding using IgG- or IgM-opsonized bacterial particles revealed a higher degree of binding in NOD mice compared with B6. We hypothesize that this novel Ig-binding receptor is part of the normal immune system function. The aberrant function in the NOD mouse could contribute to the development of Type 1 diabetes by altering normal B cell functions such as activation, IC transportation and B cell homeostasis. PMID- 19299624 TI - The effects of c-Abl mutation on developing B cell differentiation and survival. AB - c-Abl is a widely expressed Src family protein tyrosine kinase that is activated by chromosomal translocation in certain human leukemias. While shown in various experimental systems to regulate cell division and stress responses, its biological functions remain poorly understood. Although expressed at similar levels throughout B cell development, we found that the fraction of phosphorylated, active c-Abl peaks at the pro-B stage. We went on to perform a detailed analysis of B cell development in c-Abl-deficient mice. We confirmed a striking but variable decrease in pro- and pre-B cell numbers, a decrease in pre B cell growth and an increase in pre-B cell apoptosis. This phenotype was not rescued by transgenic expression of a functional IgHC transgene and only partially rescued by the anti-apoptosis gene Bcl-x. Unlike their wild-type counterparts, c-Abl-deficient pre-B cells show a defect in Ca(2+) flux upon cross linking of CD19, a co-receptor known to be involved in pre-B cell receptor signaling and failed to express CD25 on the cell surface. Despite these pre-B cell-signaling defects, selection for in-frame heavy-chain rearrangements was intact in the mutant mice. Remarkably, we were able to rescue the proliferative defect by culturing cells in vitro with large amounts of rIL-7. We conclude that c-Abl is required for normal B cell differentiation and survival. PMID- 19299626 TI - Axin expression in thymic stromal cells contributes to an age-related increase in thymic adiposity and is associated with reduced thymopoiesis independently of ghrelin signaling. AB - The adipocytes are the predominant cell types that constitute the bulk of the thymic microenvironment by the fifth decade of life in healthy humans. An age related increase in thymic adiposity is associated with reduced thymopoiesis and compromised immune surveillance in the elderly. However, the mechanisms regulating the generation of intrathymic adipocytes during aging remain to be elucidated. Here, we report that the CD45- thymic stromal cells (TSCs) are amenable to adipogenesis. We identified that the Wnt inhibitor axin is expressed in the lymphoid as well as stromal cells of the thymus with increased expression in CD45- TSCs of older mice. Knockdown of axin by RNA interference in CD45- primary TSCs led to a marked reduction in adipogenesis with significantly lower expression of adipogenic transcripts peroxisome proliferator-activated receptor 2 (PPAR), adipocyte fatty acid-binding protein (aP2), and perilipin. Age-related elevated axin expression was increased specifically in thymic fibroblasts and medullary thymic epithelial cells (TECs) but not in the cortical TEC or CD45+ cells. Consistent with a role of axin in promoting thymic adipogenesis, axin expression was also colocalized with lipid-expressing adipogenic cells in aging thymus. The prolongevity intervention, caloric restriction (CR), prevented the age-related increase in axin and the adipogenic cell in the thymus together with increase in thymic output. We have recently demonstrated that CR induces ghrelin, which can partially reverse thymic involution. Here, we show that axin expression is not affected by ablation of ghrelin receptors in aging mice, suggesting a ghrelin-independent mechanism for regulation of axin. Our data are consistent with the hypothesis that blocking the specific proadipogenic signals in the thymus may complement the present approaches to rejuvenate thymic function during aging. PMID- 19299627 TI - Towards safer carotid artery stenting: a scoring system for anatomic suitability. AB - BACKGROUNDS AND PURPOSE: To develop a scoring system to facilitate case selection for carotid artery stenting based on anatomic features. METHODS: Twelve experts comprising a multinational and multispecialty panel were convened. Delphi consensus methodology was applied over 4 "rounds" involving emailed questionnaires, private decision-making, structured interaction and explicit aggregation. In round 1 panelists proposed individual anatomic features that were considered relevant during carotid artery stenting. In round 2 each criterion was scored from 1 (straightforward) to 9 (difficult). Round 3 involved removing some factors based on individual scores to reduce the number of subsequent combination anatomies. The final round involved scoring 96 combination anatomies (representing a "full factorial" design) plus a dichotomous response, ie, whether carotid artery stenting should or should not be advised for a "novice." RESULTS: There were 1164 responses, providing a score for 12 individual anatomic features and for 96 combinations anatomies with good level of agreement between panelists. After derivation of mean (and standard deviation) of the cutting scores for 1152 yes/no responses a scoring system for combination anatomy was produced, comprising broad agreement bands presented as traffic light colors: red for particularly difficult anatomy, amber for moderate difficulty and green for lesser difficulty. CONCLUSIONS: A scoring system has been developed, based on objective expert consensus, which can be used to categorise expected difficulty of carotid artery stenting and aid case selection. PMID- 19299628 TI - Methazolamide and melatonin inhibit mitochondrial cytochrome C release and are neuroprotective in experimental models of ischemic injury. AB - BACKGROUND AND PURPOSE: The identification of a neuroprotective drug for stroke remains elusive. Given that mitochondria play a key role both in maintaining cellular energetic homeostasis and in triggering the activation of cell death pathways, we evaluated the efficacy of newly identified inhibitors of cytochrome c release in hypoxia/ischemia induced cell death. We demonstrate that methazolamide and melatonin are protective in cellular and in vivo models of neuronal hypoxia. METHODS: The effects of methazolamide and melatonin were tested in oxygen/glucose deprivation-induced death of primary cerebrocortical neurons. Mitochondrial membrane potential, release of apoptogenic mitochondrial factors, pro-IL-1beta processing, and activation of caspase -1 and -3 were evaluated. Methazolamide and melatonin were also studied in a middle cerebral artery occlusion mouse model. Infarct volume, neurological function, and biochemical events were examined in the absence or presence of the 2 drugs. RESULTS: Methazolamide and melatonin inhibit oxygen/glucose deprivation-induced cell death, loss of mitochondrial membrane potential, release of mitochondrial factors, pro-IL-1beta processing, and activation of caspase-1 and -3 in primary cerebrocortical neurons. Furthermore, they decrease infarct size and improve neurological scores after middle cerebral artery occlusion in mice. CONCLUSIONS: We demonstrate that methazolamide and melatonin are neuroprotective against cerebral ischemia and provide evidence of the effectiveness of a mitochondrial based drug screen in identifying neuroprotective drugs. Given the proven human safety of melatonin and methazolamide, and their ability to cross the blood-brain barrier, these drugs are attractive as potential novel therapies for ischemic injury. PMID- 19299629 TI - Sequencing of TGF-beta pathway genes in familial cases of intracranial aneurysm. AB - BACKGROUND AND PURPOSE: Familial aggregation of intracranial aneurysms (IA) strongly suggests a genetic contribution to pathogenesis. However, genetic risk factors have yet to be defined. For families affected by aortic aneurysms, specific gene variants have been identified, many affecting the receptors to transforming growth factor-beta (TGF-beta). In recent work, we found that aortic and intracranial aneurysms may share a common genetic basis in some families. We hypothesized, therefore, that mutations in TGF-beta receptors might also play a role in IA pathogenesis. METHODS: To identify genetic variants in TGF-beta and its receptors, TGFB1, TGFBR1, TGFBR2, ACVR1, TGFBR3, and ENG were directly sequenced in 44 unrelated patients with familial IA. Novel variants were confirmed by restriction digestion analyses, and allele frequencies were analyzed in cases versus individuals without known intracranial disease. Similarly, allele frequencies of a subset of known SNPs in each gene were also analyzed for association with IA. RESULTS: No mutations were found in TGFB1, TGFBR1, TGFBR2, or ACVR1. Novel variants identified in ENG (p.A60E) and TGFBR3 (p.W112R) were not detected in at least 892 reference chromosomes. ENG p.A60E showed significant association with familial IA in case-control studies (P=0.0080). No association with IA could be found for any of the known polymorphisms tested. CONCLUSIONS: Mutations in TGF-beta receptor genes are not a major cause of IA. However, we identified rare variants in ENG and TGFBR3 that may be important for IA pathogenesis in a subset of families. PMID- 19299630 TI - Relation between cerebral perfusion territories and location of cerebral infarcts. AB - BACKGROUND AND PURPOSE: The perfusion territories of the brain-feeding arteries are difficult to assess in vivo and therefore standard cerebral perfusion territory templates are often used to determine the relation between cerebral infarcts and the feeding vasculature. In the present study, we compared this infarct classification, using standard templates, with the individualized depiction of cerebral perfusion territories on MRI. METHODS: The ethics committee of our institution approved the study protocol. A total of 159 patients (92 male, 67 female; mean age, 58.9 years) with first-time clinical symptoms of cerebral ischemia were included in the study. Diffusion-weighted imaging was used for depiction of the area of ischemia and the perfusion territories of the left internal carotid artery, right internal carotid artery, and vertebrobasilar arteries were visualized with territorial arterial spin labeling MRI. Infarct locations with respect to cerebral perfusion territories were evaluated with and without territorial arterial spin labeling MRI images. RESULTS: In 92% of the patients, the territorial arterial spin labeling images were of diagnostic quality. One hundred thirty-six patients showed areas of ischemia on diffusion weighted images. The additional information from the territorial arterial spin labeling images changed the classification in 11% of the cortical or border zone infarcts (6 of 56), whereas no territorial changes were observed in lacunar, periventricular, cerebellar, and brainstem infarcts. CONCLUSIONS: The diagnostic information provided by perfusion territory imaging in patients with stroke is valuable for the classification of cortical and border zone infarcts, whereas no change of the textbook-based classification was observed for other infarct types. PMID- 19299631 TI - Abciximab is a safe rescue therapy in thromboembolic events complicating cerebral aneurysm coil embolization: single center experience in 42 cases and review of the literature. AB - BACKGROUND AND PURPOSE: The purpose of this study was to estimate the safety and efficacy of abciximab treatment in combination with prophylactic heparin, acetylsalicylic acid (ASA), and clopidogrel application in cases of thrombus formation complicating endovascular coil embolization in cerebral aneurysms. METHODS: Thromboembolic incidents during endovascular management of 515 consecutive cerebral aneurysms were observed in 48 cases (9.3%). Eight incidents were observed during embolization of incidental aneurysms (8/174; 4.6%, 95% CI: 2.0 to 8.9%). All patients underwent anticoagulation with heparin and platelet inhibition with ASA during treatment procedure. In addition, clopidogrel orally was applied 3 days preoperatively in patients with incidental aneurysms. In case of thrombus formation, glycoprotein IIb-IIIa inhibitor abciximab was given in 42 cases. No coagulation-effective rescue treatment was conducted in 5 cases. One patient was treated with r-tPA. End points were infarction on follow-up cranial CT and the rate of intracranial hemorrhages. RESULTS: No infarcts on follow-up CT were observed after treatment with abciximab in 29/42 patients (69.0%, 95% CI: 52.9 to 82.4%). No coagulant rescue therapy was applied in 5 patients because of a small nonocclusive thrombus or good collateral blood supply, showing consecutive infarction on follow-up CT in 3 cases as did the 1 patient treated with r-tPA. No periprocedural bleedings or rebleedings were observed in any case. CONCLUSIONS: Abciximab was safe as rescue treatment in cases of thrombus formation during endovascular aneurysm coiling. In our study the use of Abciximab, in combination with prophylactic anticoagulation and antiaggregation, does not lead to additional intracranial hemorrhages or any extracranial bleeding complications. PMID- 19299632 TI - Relationships between cerebral perfusion and reversibility of acute diffusion lesions in DEFUSE: insights from RADAR. AB - BACKGROUND AND PURPOSE: Acute ischemic lesions with restricted diffusion can resolve after early recanalization. The impact of superimposed perfusion abnormalities on the fate of acute diffusion lesions is unclear. METHODS: Data were obtained from DEFUSE, a prospective multicenter study of patients treated with IV tPA 3 to 6 hours after stroke onset. Thirty-two patients with baseline diffusion and perfusion lesions and 30 day FLAIR scans were coregistered. The acute diffusion lesion was divided into 3 regions according to the Tmax delay of the superimposed perfusion lesion: normal baseline perfusion; mild-moderately hypoperfused (2 s8 s). The reversal rate was calculated as the percentage of the acute diffusion lesion that did not overlap with the final infarct on 30-day FLAIR. Diffusion reversal rates were compared based on whether a favorable clinical response occurred and whether early recanalization was achieved. RESULTS: On average, 54% of the acute diffusion lesion volume had normal perfusion. Diffusion reversal rates were significantly increased among cases with favorable clinical response and in patients with early recanalization, especially in regions with normal baseline perfusion. The portion of the diffusion lesion with normal perfusion had significantly higher mean apparent diffusion coefficient values and reversal rates. CONCLUSIONS: Acute ischemic lesions with restricted diffusion are most likely to recover if reperfusion occurs within 6 hours of symptom onset, and reversibility is associated with early recanalization and favorable clinical outcome. We propose the term RADAR (Reversible Acute Diffusion lesion Already Reperfused) to describe regions of acute restricted diffusion with normal perfusion. PMID- 19299633 TI - Combined aspirin plus warfarin: recent evidence and residual questions. PMID- 19299634 TI - Does the combination of warfarin and aspirin have a place in secondary stroke prevention? No. PMID- 19299635 TI - Does the combination of warfarin and aspirin have a place in secondary stroke prevention? Yes. PMID- 19299637 TI - Is plasma fibrinogen useful in evaluating ischemic stroke patients?: why, how, and when. PMID- 19299636 TI - Results of the MRI substudy of the intravenous magnesium efficacy in stroke trial. AB - BACKGROUND AND PURPOSE: Although magnesium is neuroprotective in animal stroke models, no clinical benefit was confirmed in the Intravenous Magnesium Efficacy in Stroke (IMAGES) trial of acute stroke patients. The Magnetic Resonance in IMAGES (MR IMAGES) substudy investigated the effects of magnesium on the imaging surrogate outcome of infarct growth. METHODS: IMAGES trial patients in participating centers were randomized to receive either intravenous magnesium or placebo within 12 hours of stroke onset. Infarct growth was defined as volume difference between baseline diffusion-weighted imaging and day 90 fluid attenuated inversion recovery image lesions. Patients who died were imputed the largest infarct growth observed. RESULTS: Among the 90 patients included in the primary analysis, there was no difference in infarct growth (median absolute growth, P=0.639; median percentage growth, P=0.616; proportion with any growth, P=0.212) between the 46 treated with magnesium and 44 with placebo. Infarct growth correlated with NIHSS score change from baseline to day 90. There was a trend showing baseline serum glucose correlated with infarct growth with magnesium treatment, but not in the placebo group. The mismatch frequency was reduced from 73% to 47% by increasing the mismatch threshold from >20% to >100% of core volume. CONCLUSIONS: Infarct growth, confirmed here as a surrogate for clinical progression, was similar between magnesium and placebo treatment, paralleling the main IMAGES trial clinical outcomes. Glucose was a covariate for infarct growth with magnesium treatment. A more stringent mismatch threshold to define penumbra more appropriately would have excluded half of the patients in this 12-hour time window stroke study. PMID- 19299638 TI - Serum cholesterol LDL and 90-day mortality in patients with intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Prognostic significance of low-density lipoprotein cholesterol (LDL-C) in intracranial hemorrhage (ICH) is unclear. The objective of this study was to determine the association between LDL-C and mortality in ICH. METHODS: Consecutive patients (n=88) presenting with ICH were included in the study. Lipid profile was obtained during the first hours after admission. We analyzed the impact of LDL-C on 90-day mortality using the Hazard Rate (HR) crude, analysis crude for trend by Mantel-Haenszel Test, Multiple Cox Proportional Hazards model, and analysis of survival curves. Association between LDL-C and severity markers of ICH were explored using Spearman correlation coefficient. RESULTS: Low LDL-C levels were independently associated with death after intracranial hemorrhage (HR=3.07 (95% CI:1.04 to 9.02; P=0.042) in multivariable analysis after controlling for confounding factors. Analysis for trend showed a significant association (Xt=-2.144; P=0.032) by Mantel-Haenszel Test. Spearman analysis showed no correlation between LDL-C and variables that are markers of ICH severity: NIH score (r=-0.091; P=0.400), GCS score (r=0.136; P=0.207), ICH volume (r=0.140; P=0.192), and length of stay (r=-0.111; P=0.308). CONCLUSIONS: Low levels of LDL-C are independently associated with an increased risk of death in patients with brain hemorrhage. We have not found evidences that the levels of LDL-C can act as a biological marker of severity. PMID- 19299639 TI - Differential infraslow (<0.1 Hz) cortical activations in the affected and unaffected hemispheres from patients with subacute stroke demonstrated by noninvasive DC-magnetoencephalography. AB - BACKGROUND AND PURPOSE: Sustained mass depolarization of neurons, termed cortical spreading depolarization, is one electrophysiological correlate of the ischemic injury of neurons. Cortical spreading depolarizations spread in the gray matter at a rate of approximately 3 mm/min and are associated with large infraslow extracellular potential changes (<0.05 Hz). Moreover, smaller infraslow potential changes accompany functional activation and might help to assess neuronal repair after stroke. The objective of the present pilot study was to investigate whether it is feasible to apply noninvasive near-DC-magnetoencephalography to detect and monitor infraslow field changes in patients with acute stroke. METHODS: A simple motor condition was used to induce physiological cortical infraslow field changes. Five patients in a subacute state after ischemic stroke performed self paced simple finger movements (30-second periods of finger movements, always separated by 30-second periods of rest, for a total of 15 minutes). Near-DC magnetoencephalography signals were recorded over the contralateral primary motor cortex for the affected and unaffected hemisphere, respectively. RESULTS: In all patients, the time courses of the contralateral cortical field amplitudes in the infraslow frequency range followed closely the motor task cycles revealing statistically significant differences between finger movement and rest periods. In 4 of 5 patients, infraslow field amplitudes were significantly stronger over the unaffected hemisphere compared with the affected hemisphere. CONCLUSIONS: This study demonstrates that cortical infraslow activity can be recorded noninvasively in patients in the subacute state after ischemic stroke. It is suggested that near-DC-magnetoencephalography is a promising tool to also detect cortical spreading depolarization noninvasively. PMID- 19299640 TI - Reduction in incident stroke risk with vigorous physical activity: evidence from 7.7-year follow-up of the national runners' health study. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess the dose-response relationship between vigorous physical activity (running distance, km/d) and the participant-reported physician-diagnosed stroke. METHODS: Age-adjusted survival analysis of 29 279 men and 12 123 women followed prospectively for 7.7 years. RESULTS: One hundred men and 19 women reported incident strokes. Per km/d run, the age- and smoking-adjusted risk for stroke decreased 12% in men (P=0.0007), and 11% in men and women combined (P=0.001), which remained significant when further adjusted for baseline diabetes, hypercholesterolemia, hypertension, and BMI (8% and 7% reduction per km/d run, respectively, P=0.03). Men and women who ran >or=2 km/d (ie, exceeded the recommended AHA/CDC and NIH guideline activity level) had significantly lower risk than those who ran less (P=0.05), and those who ran >or=4 km/d had significantly lower risk than those who ran 2 to 3.9 km/d (P=0.02). Men and women who ran >or=8 km/d were at 60% lower risk than those who ran <2 km/d (P=0.00). CONCLUSIONS: The risk for incident stroke is substantially reduced in those who exceed the guideline physical activity level, which cannot be attributed to less hypertension, diabetes, hypercholesterolemia, or body weight. PMID- 19299641 TI - Regional implementation of the stroke systems of care model: recommendations of the northeast cerebrovascular consortium. AB - BACKGROUND AND PURPOSE: The Northeast Cerebrovascular Consortium was established to examine regional disparities and recommend strategies to improve stroke care based on the Stroke Systems of Care Model. METHODS: An annual summit was first held in 2006, bringing together public health officials, researchers, physicians, nurses, health professionals, state legislators, and advocacy organizations. Best practices and evidence-based interventions within each of the Stroke Systems of Care Model components were presented. Six writing groups were tasked with cataloging each state's current activities and identifying goals for the region. RESULTS: There were significant variations in the delivery of stroke care, particularly in urban versus rural areas, as evidenced by the availability of designated stroke centers and neurologists, and stroke-related death rates. Recommendations to address variations in care delivery included the use of a common stroke data collection system, unified community education criteria, improvements to emergency medical services dispatch and training, adoption of prehospital care measures, creation of a web-based central repository of acute stroke protocols and order sets, a regional atlas of stroke resources and capabilities, a stroke patient "report card" to promote adherence to secondary prevention strategies, and explicit standards for rehabilitation services. CONCLUSIONS: Significant disparities in the delivery of stroke care across the 8 state-region have been identified. Northeast Cerebrovascular Consortium demonstrates that multistate regional collaboration is a viable process for developing specific regional recommendations to address those disparities. Northeast Cerebrovascular Consortium is assessing the usefulness of the Stroke Systems of Care Model as a framework for implementing a regional approach to stroke across the continuum of care. PMID- 19299642 TI - Hyperfibrinogenemia and functional outcome from acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Epidemiological studies have found strong correlations between elevated plasma fibrinogen levels and both ischemic stroke incidence and stroke mortality. Little is known about the influence of fibrinogen levels on functional stroke outcome. METHODS: Placebo data from the Stroke Treatment with Ancrod Trial (STAT) and European Stroke Treatment with Ancrod Trial (ESTAT) were analyzed. Fibrinogen levels were determined within 3 hours (STAT) or 6 hours (ESTAT) of stroke onset and at preset intervals throughout 5 days of intravenous infusions. Barthel Index scores at 90 days quantified functional outcomes. The association between initial fibrinogen levels and functional outcomes was evaluated using a multiple logistic regression analysis. RESULTS: Fibrinogen levels increased gradually over the first 24 hours from a pretreatment median value of 340 mg/dL to a 24-hour median value of 376 mg/dL. In a univariate analysis, the proportion of patients with good functional outcome decreased with increasing quartiles of initial fibrinogen levels in both STAT (36.0% to 26.2%) and ESTAT (53.8% to 24.8%). In a multifactorial analysis, the same trend was observed. Patients with initial fibrinogen levels <450 mg/dL had better outcomes in both studies; the difference (42.0% versus 21.6%) was significant in ESTAT (P=0.0006), even when corrected for age and initial stroke severity. CONCLUSIONS: The independent association of higher initial fibrinogen levels with poor outcome needs to be verified using a larger acute stroke dataset. Even in the present small populations, the apparent association of these 2 variables suggests that treatments designed to reduce fibrinogen levels could potentially be important in treating acute ischemic stroke. PMID- 19299643 TI - Biochemical and mechanical dysfunction in a mouse model of desmin-related myopathy. AB - An R120G mutation in alphaB-crystallin (CryAB(R120G)) causes desmin-related myopathy (DRM). In mice with cardiomyocyte-specific expression of the mutation, CryAB(R120G)-mediated DRM is characterized by CryAB and desmin accumulations within cardiac muscle, mitochondrial deficiencies, activation of apoptosis, and heart failure (HF). Excessive production of reactive oxygen species (ROS) is often a hallmark of HF and treatment with antioxidants can sometimes prevent the progression of HF in terms of contractile dysfunction and cardiomyocyte survival. It is unknown whether blockade of ROS is beneficial for protein misfolding diseases such as DRM. We addressed this question by blocking the activity of xanthine oxidase (XO), a superoxide-generating enzyme that is upregulated in our model of DRM. The XO inhibitor oxypurinol was administered to CryAB(R120G) mice for a period of 1 or 3 months. Mitochondrial function was dramatically improved in treated animals in terms of complex I activity and conservation of mitochondrial membrane potential. Oxypurinol also largely restored normal mitochondrial morphology. Surprisingly, however, cardiac contractile function and cardiac compliance were unimproved, indicating that the contractile deficit might be independent of mitochondrial dysfunction and the initiation of apoptosis. Using magnetic bead microrheology at the single cardiomyocyte level, we demonstrated that sarcomeric disarray and accumulation of the physical aggregates resulted in significant changes in the cytoskeletal mechanical properties in the CryAB(R120G) cardiomyocytes. Our findings indicate that oxypurinol treatment largely prevented mitochondrial deficiency in DRM but that contractility was not improved because of mechanical deficits in passive cytoskeletal stiffness. PMID- 19299644 TI - T cell-mediated inflammation in adipose tissue does not cause insulin resistance in hyperlipidemic mice. AB - Obesity is associated with chronic inflammation in adipose tissue. Proinflammatory cytokines including tumor necrosis factor-alpha and interleukin-6 secreted by adipose tissue during the metabolic syndrome are proposed to cause local and general insulin resistance and promote development of type 2 diabetes. We have used a compound mutant mouse, Apoe(-/-)xCD4dnTGFbR, with dysregulation of T-cell activation, excessive production of proinflammatory cytokines, hyperlipidemia, and atherosclerosis, to dissect the role of inflammation in adipose tissue metabolism. These mice are lean, which avoids confounding effects of concomitant obesity. Expression and secretion of a set of proinflammatory factors including tumor necrosis factor-alpha, interferon-gamma, and monocyte chemoattractant protein-1 was increased in adipose tissue of Apoe(-/-)xCD4dnTGFbR mice, as was the enzyme 11beta-hydroxysteroid dehydrogenase type 1, which converts cortisone to bioactive cortisol. Interleukin-6, which has an inhibitory glucocorticoid response element in its promoter, was not upregulated. In spite of intense local inflammation, insulin sensitivity was not impaired in adipose tissue of Apoe(-/-)xCD4dnTGFbR mice unless exogenous interleukin-6 was administered. In conclusion, T-cell activation causes inflammation in adipose tissue but does not lead to insulin resistance in this tissue in the absence of interleukin-6. PMID- 19299645 TI - Inducible expression of active protein phosphatase-1 inhibitor-1 enhances basal cardiac function and protects against ischemia/reperfusion injury. AB - Ischemic heart disease, which remains the leading cause of morbidity and mortality in the Western world, is invariably characterized by impaired cardiac function and disturbed Ca(2+) homeostasis. Because enhanced inhibitor-1 (I-1) activity has been suggested to preserve Ca(2+) cycling, we sought to define whether increases in I-1 activity in the adult heart may ameliorate contractile dysfunction and cellular injury in the face of an ischemic insult. To this end, we generated an inducible transgenic mouse model that enabled temporally controlled expression of active I-1 (T35D). Active I-1 expression in the adult heart elicited significant enhancement of contractile function, associated with preferential phospholamban phosphorylation and enhanced sarcoplasmic reticulum Ca(2+)-transport. Further phosphoproteomic analysis revealed alterations in proteins associated with energy production and protein synthesis, possibly to support the increased metabolic demands of the hyperdynamic hearts. Importantly, on ischemia/reperfusion-induced injury, active I-1 expression augmented contractile function and recovery. Further examination revealed that the infarct region and apoptotic as well as necrotic injuries were significantly attenuated by enhanced I-1 activity. These cardioprotective effects were associated with suppression of the endoplasmic reticulum stress response. The present findings indicate that increased I-1 activity in the adult heart enhances Ca(2+) cycling and improves mechanical recovery, as well as cell survival after an ischemic insult, suggesting that active I-1 may represent a potential therapeutic strategy in myocardial infarction. PMID- 19299646 TI - Endothelium-dependent cerebral artery dilation mediated by TRPA1 and Ca2+ Activated K+ channels. AB - Although it is well established that changes in endothelial intracellular [Ca(2+)] regulate endothelium-dependent vasodilatory pathways, the molecular identities of the ion channels responsible for Ca(2+) influx in these cells are not clearly defined. The sole member of the ankyrin (A) transient receptor potential (TRP) subfamily, TRPA1, is a Ca(2+)-permeable nonselective cation channel activated by electrophilic compounds such as acrolein (tear gas), allicin (garlic), and allyl isothiocyanate (AITC) (mustard oil). The present study examines the hypothesis that Ca(2+) influx via TRPA1 causes endothelium-dependent vasodilation. The effects of TRPA1 activity on vascular tone were examined using isolated, pressurized cerebral arteries. AITC induced concentration-dependent dilation of pressurized vessels with myogenic tone that was accompanied by a corresponding decrease in smooth muscle intracellular [Ca(2+)]. AITC-induced dilation was attenuated by disruption of the endothelium and when the TRPA1 channel blocker HC-030031 was present in the arterial lumen. TRPA1 channels were found to be present in native endothelial cells, localized to endothelial cell membrane projections proximal to vascular smooth muscle cells. AITC-induced dilation was insensitive to nitric oxide synthase or cyclooxygenase inhibition but was blocked by luminal administration of the small and intermediate conductance Ca(2+)-activated K(+) channel blockers apamin and TRAM34. BaCl(2), a blocker of inwardly rectifying K(+) channels, also inhibited AITC-induced dilation. AITC-induced smooth muscle cell hyperpolarization was blocked by apamin and TRAM34. We conclude that Ca(2+) influx via endothelial TRPA1 channels elicits vasodilation of cerebral arteries by a mechanism involving endothelial cell Ca(2+)-activated K(+) channels and inwardly rectifying K(+) channels in arterial myocytes. PMID- 19299647 TI - Acquisition of brain Na sensitivity contributes to salt-induced sympathoexcitation and cardiac dysfunction in mice with pressure overload. AB - In animal models of salt-sensitive hypertension, high salt augments sympathetic outflow via central mechanisms. It is not known, however, whether pressure overload affects salt sensitivity, thereby modifying central sympathetic outflow and cardiac function. We induced left ventricular hypertrophy with aortic banding in mice. Four weeks after aortic banding (AB-4), the left ventricle wall thickness was increased without changing the percentage fractional shortening. AB 4 mice were then fed either a high-salt (8%) diet or regular-salt diet for additional 4 weeks. Cardiac dysfunction, wall thickness, and 24-hour urinary catecholamine excretion were increased with high-salt diet compared with regular salt diet. We then examined brain Na sensitivity. Intracerebroventricular infusion of high-Na (0.2 mol/L) artificial cerebrospinal fluid into AB-4 mice and mice Sham-4 increased urinary catecholamine excretion, arterial pressure, and heart rate more in AB-4 mice than in Sham-4 mice. Intracerebroventricular infusion of an epithelial Na channel blocker (benzamil) into mice with high-salt diet significantly decreased urinary catecholamine excretion and improved cardiac function. Infusion of either an angiotensin II type 1 receptor blocker or a Rho kinase inhibitor also attenuated the salt-induced sympathetic hyperactivation and cardiac dysfunction in mice with high-salt diet. The levels of angiotensin II type 1 receptor and phosphorylated moesin, a substrate of Rho-kinase, were significantly greater in AB-4 mice than in Sham-4 mice. These results suggest that mice with pressure overload acquire brain Na sensitivity because of the activation of epithelial Na channel via Rho-kinase and angiotensin II, and this mechanism contributes to salt-induced sympathetic hyperactivation, further pressure overload, and cardiac dysfunction. PMID- 19299648 TI - Trigeminal neuralgia due to anterior inferior cerebellar artery loop: a case report. AB - Trigeminal neuralgia is uncommon in children. Idiopathic type is less often seen than the secondary or symptomatic type. We report a 6-year-old girl with trigeminal neuralgia. Magnetic resonance imaging revealed an arterial loop at the exit of the trigeminal nerve. PMID- 19299649 TI - Delirium in children with HIV/AIDS. AB - Delirium in children has received little attention from researchers until very recently, despite being a frequently encountered clinical problem, particularly in the context of human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS). In this article, we report 3 cases of severe hyperactive delirium occurring in children with advanced HIV/AIDS in the context of probable underlying HIV encephalopathy and complex drug-drug interactions. We also present a hypothesis in relation to HIV-associated delirium as a potential neuropsychiatric manifestation of the immune reconstitution inflammatory syndrome in children commenced on highly active antiretroviral therapy. PMID- 19299650 TI - Desensitization to type I interferon in HIV-1 infection correlates with markers of immune activation and disease progression. AB - Type I interferon (IFNalpha/beta) plays a complex role in HIV-1 infection and has been proposed alternately to have roles in either disease protection or progression. Although IFNalpha/beta plays crucial roles in regulating monocytes and dendritic cells, responsiveness of these cells to IFNalpha/beta in HIV-1 infection is poorly understood. We report significant defects in IFNalpha/beta receptor (IFNalpha/betaR) expression, IFNalpha signaling, and IFNalpha-induced gene expression in monocytes from HIV-1-infected subjects. IFNalpha/betaR expression correlated directly with CD4+ T-cell count and inversely with HIV-1 RNA level and expression of CD38 by memory (CD45RO+) CD8+ T cells, a measure of pathologic immune activation in HIV-1 infection associated with disease progression. In addition, monocytes from HIV-1-infected persons showed diminished responses to IFNalpha, including decreased induction of phosphorylated STAT1 and the classical interferon-stimulated gene produces MxA and OAS. These IFNalpha responses were decreased regardless of IFNalpha/betaR expression, suggesting that regulation of intracellular signaling may contribute to unresponsiveness to IFNalpha/beta in HIV-1 disease. Defective monocyte responses to IFNalpha/beta may play an important role in the pathogenesis of HIV-1 infection, and decreased IFNalpha/betaR expression may serve as a novel marker of disease progression. PMID- 19299651 TI - Unraveling ALK signaling through phosphoproteomics. PMID- 19299652 TI - The expanding universe of the basophil. PMID- 19299653 TI - To give and take - life of a platelet. PMID- 19299654 TI - Intriguing response to azacitidine in a patient with juvenile myelomonocytic leukemia and monosomy 7. PMID- 19299655 TI - Etanercept for treatment of idiopathic pneumonia syndrome after allogeneic hematopoietic stem cell transplantation. PMID- 19299657 TI - The dark ages illuminated: the "humorless" remedies of pseudo-apuleius. PMID- 19299658 TI - Visions of drug discovery. Cimetidine (Tagamet). PMID- 19299659 TI - Short and sweet: evolution of a small glycopeptide from a bladder disorder to an anticancer lead. AB - Glycopeptides are a class of molecules that comprise two distinct families of biologically important scaffolds, peptides and oligosaccharides, each playing important roles in cellular communication and signaling. Rarely are small, endogenous secreted glycopeptides found that have significant impact on the progression of a specific disease state, but such is the case for the antiproliferative factor (APF) found in the urine and tissue of patients with the poorly understood bladder diseases collectively referred to as interstitial cystitis (IC). APF is a 9-mer peptide containing a sialylated O-linked trisaccharide glycan attached to the N-terminal threonine. APF dramatically inhibits normal bladder cell proliferation and is thought to cause some of the characteristic pathological changes in the bladder of IC patients. Importantly, APF also potently inhibits the growth of certain tumor cells. The details of the cellular receptors to which APF interacts, and the structural features that are critical for its potency are now beginning to unfold. This interesting molecule is a powerful model for the design of new treatments and diagnostic tests for IC, as well as an unprecedented lead agent for novel anticancer drug design. PMID- 19299660 TI - The Outcome of Selenium and Vitamin E Cancer Prevention Trial (SELECT) reveals the need for better understanding of selenium biology. AB - The recently completed Selenium and Vitamin E Cancer Prevention Trial (SELECT) was one of the largest human cancer prevention trials ever undertaken. Its purpose was to assess the role of selenium and vitamin E in prostate cancer prevention, but SELECT found no decline in prostate cancer. Comparison of this study to other clinical trials involving selenium and to the results of animal studies suggests that the source of the selenium supplement, L-selenomethionine, and the relatively high initial levels of selenium in the enrolled men may have contributed to this outcome. Further analysis of the clinical and animal data highlights the need for mechanistic studies to better understand selenium biology in order to target dietary selenium to appropriate subsets of the human population: those individuals most likely to benefit from this micronutrient. PMID- 19299661 TI - Fragment-based ligand discovery. AB - From home building and decor to mass production, modular design is a standard feature of the modern age. The concept also promises to define drug discovery efforts in the near future, as a wide range of methodologies, from NMR to X-ray crystallography, are being adapted to high-throughput platforms. In particular, "fragment-based ligand discovery" describes the laboratory-driven evolution of drugs from libraries of chemical building blocks. "Evolution" is an apt word for the process, as a wide array of methods are used to define how compound fragments can be best fit into the binding sites of medically relevant target biomolecules. A number of compounds that evolved from fragments have entered the clinic, and the approach is increasingly accepted as an additional route to identifying new hit compounds in pharmaceutical discovery and inhibitor design. PMID- 19299662 TI - Low-dose aspirin, coxibs, and other NSAIDS: a clinical mosaic emerges. AB - Aspirin has been a commercial drug for over a century, although for most of this history, an understanding of its mechanism of action, as an inhibitor of cyclooxygenase (COX) activity and thus of prostanoid synthesis, was lacking. Over the past fifty years, a large number of other nonsteroidal antiinflammatory drugs (NSAIDs) have been developed, and a much deeper understanding of inflammation and prostanoid action has emerged. Indeed, a new class of selective inhibitors of the cyclooxygenase-2 isozyme was introduced, about ten years ago, and these so-called coxibs quickly became regarded as preferable, in certain clinical contexts, to avoid side effects associated with the use of aspirin and previously developed NSAIDs. This regard for coxibs has been challenged, sometimes infamously, as cardiovascular events associated with coxib use have become apparent. A variety of clinical trials have led to seemingly conflicting data concerning the roles of COX-1 and COX-2, and the implications of their relative inhibition, in cardiovascular health and disease. In this Review, the authors offer an assessment of drug pharmacokinetics and enzyme physiology that reconciles cardiovascular appraisals from a wide array of clinical data. PMID- 19299663 TI - The transforming power of art. PMID- 19299664 TI - Measuring the value of public health systems. PMID- 19299665 TI - Abortion among couples in rural Bangladesh. PMID- 19299666 TI - Nurses' utilization and perception of the community/public health nursing credential. AB - OBJECTIVES: I explored the underutilization of the community/public health nursing (C/PHN) credential by examining the individual characteristics of public health nurses, the value these nurses perceive for certification, the barriers they perceive to obtaining or maintaining a C/PHN credential, and their credential status. METHODS: I surveyed a national sample of 655 public health nurses regarding this more than 20-year-old credential. I analyzed variables related to perceived value, barriers, and characteristics of public health nurses. RESULTS: The perceived value of credentialing did not differ among public health nurses relative to whether they had ever had a C/PHN credential. The C/PHN credential, however, was obtained significantly more often by public health nurses in academic settings than by those working in practice settings. CONCLUSIONS: The C/PHN credential appears to be disproportionately underutilized and unknown to public health nurses in the practice community. Findings suggest, however, that utilization could be improved by increasing the credential's visibility outside of academic environments and by establishing system-level changes that provide external recognition, such as salary increases and career advancement, for specialty credentials. PMID- 19299667 TI - Clinical and demographic factors associated with homelessness and incarceration among VA patients with bipolar disorder. AB - OBJECTIVES: We assessed the association between homelessness and incarceration in Veterans Affairs patients with bipolar disorder. METHODS: We used logistic regression to model each participant's risk of incarceration or homelessness after we controlled for known risk factors. RESULTS: Of 435 participants, 12% reported recent homelessness (within the past month), and 55% reported lifetime homelessness. Recent and lifetime incarceration rates were 2% and 55%, respectively. In multivariate models, current medication adherence (based on a 5 point scale) was independently associated with a lower risk of lifetime homelessness (odds ratio [OR] = 0.80 per point, range 0-4; 95% confidence interval [CI] = 0.66, 0.96), and lifetime incarceration increased the risk of lifetime homelessness (OR = 4.4; 95% CI = 2.8, 6.9). Recent homelessness was associated with recent incarceration (OR = 26.4; 95% CI = 5.2, 133.4). Lifetime incarceration was associated with current substance use (OR = 2.6; 95% CI = 2.7, 6.7) after control for lifetime homelessness (OR = 4.2; 95% CI = 2.7, 6.7). CONCLUSIONS: Recent and lifetime incarceration and homelessness were strongly associated with each other. Potentially avoidable or treatable correlates included current medication nonadherence and substance use. Programs that better coordinate psychiatric and drug treatment with housing programs may reduce the cycle of incarceration, homelessness, and treatment disruption within this vulnerable patient population. PMID- 19299668 TI - Health reform redux: learning from experience and politics. AB - The 2008 presidential campaign season featured health care reform proposals. I discuss 3 approaches to health care reform and the tools for bringing about reform, such as insurance market reforms, tax credits, subsidies, individual and employer mandates, and public program expansions. I also discuss the politics of past and current health care reform efforts. Market-based reforms and mandates have been less successful than public program expansions at expanding coverage and controlling costs. New divisions among special interest groups increase the likelihood that reform efforts will succeed. Federal support for state efforts may be necessary to achieve national health care reform. History suggests that state-level success precedes national reform. History also suggests that an organized social movement for reform is necessary to overcome opposition from special interest groups. PMID- 19299669 TI - Reduction in suicide mortality following a new national alcohol policy in Slovenia: an interrupted time-series analysis. AB - OBJECTIVES: We assessed the impact on suicide mortality of a new national policy in Slovenia that limits the availability of alcohol. METHODS: We obtained monthly total, male, and female suicide counts in Slovenia between January 1997 and December 2005 and then employed autoregressive integrated moving average (ARIMA) techniques to model the effect of the alcohol policy (implemented in March 2003). RESULTS: There was a significant decrease in the total number of monthly suicides following the policy's implementation. Subsequent analyses revealed this association to be caused solely by the impact on male suicides. Specifically, there was an immediate and permanent reduction of 3.6 male suicides per month (95% confidence interval = -0.4, -6.9), or approximately 10% of the preintervention average. The policy had no statistically significant effect on female suicides. CONCLUSIONS: Our results show the effectiveness of this specific policy in reducing male suicides in Slovenia and also hint at the potential of public policy in reducing the public health burden of alcohol-related harm more generally. PMID- 19299670 TI - Preventing tobacco use among young people in India: Project MYTRI. AB - OBJECTIVES: We assessed the effectiveness of a 2-year multicomponent, school based intervention designed to reduce tobacco use rates among adolescents in an urban area of India. METHODS: Students from 32 schools in Delhi and Chennai, India, were recruited and randomly assigned to an intervention or control group. Baseline, intermediate, and outcome data were collected from 2 cohorts of 6th- and 8th-grade students in 2004; 14,063 students took part in the study and completed a survey in 2004, 2005, or 2006. The intervention consisted of behavioral classroom curricula, school posters, a parental involvement component, and peer-led activism. The main outcome measures were self-reported use of cigarettes, bidis (small hand-rolled, often flavored, cigarettes), and chewing tobacco and future intentions to smoke or use chewing tobacco. RESULTS: Findings showed that students in the intervention group were significantly less likely than were students in the control group to exhibit increases in cigarette smoking or bidi smoking over the 2-year study period. They were also less likely to intend to smoke or chew tobacco in the future. CONCLUSIONS: School-based programs similar to the intervention examined here should be considered as part of a multistrategy approach to reducing tobacco use among young people in India. PMID- 19299671 TI - Understanding the decisions and values of stakeholders in health information exchanges: experiences from Massachusetts. AB - OBJECTIVES: We studied how health information exchange systems are established by examining the decisions (and thus, indirectly, the values) of key stakeholders (health care providers) participating in a health information exchange pilot project in 3 Massachusetts communities. Our aim was to understand how these kinds of information exchanges can be made viable. METHODS: We used semistructured interviews to assess health care providers' decision-making processes in selecting technical architectures and vendors for the pilot projects to uncover their needs, expectations, and motivations. RESULTS: Our interviews indicated that, after extensive evaluations, health care providers in all 3 communities eventually selected a hybrid architecture that included a central data repository. However, the reasons for selecting this architecture varied considerably among the 3 communities, reflecting their particular values. Plans to create a community patient portal also differed across communities. CONCLUSIONS: Our findings suggest that, to become established, health information exchange efforts must foster trust, appeal to strategic interests of the medical community as a whole, and meet stakeholder expectations of benefits from quality measurements and population health interventions. If health information exchange organizations cannot address these factors, sustainability will remain precarious. PMID- 19299672 TI - The contribution of clinic-based interventions to reduce prenatal smoking prevalence among US women. AB - OBJECTIVES: We sought to estimate the effect of universal implementation of a clinic-based, psychosocial smoking cessation intervention for pregnant women. METHODS: We used data from US birth certificates (2005) and the Pregnancy Risk Assessment Monitoring System (2004) to estimate the number of women smoking at conception. To calculate the number of women eligible to receive the cessation intervention, we used estimates from the literature of the percentage of women who quit spontaneously (23%), entered prenatal care before the third trimester (96.5%), and disclosed smoking to their provider (75%). We used the pooled relative risk (RR) for continued smoking from the 2004 Cochrane Review as our measure of the intervention's effectiveness (RR = 0.94). RESULTS: We estimated that 944,240 women smoked at conception. Of these, 23.0% quit spontaneously, 6.3% quit with usual care, and an additional 3.3% quit because of the intervention, leaving 67.4% smoking throughout pregnancy. The calculated smoking prevalence in late pregnancy decreased from 16.4% to 15.6% because of the intervention. CONCLUSIONS: Universal implementation of a best-practice, clinic-based intervention would increase the total number of quitters but would not substantially reduce smoking prevalence among pregnant women. PMID- 19299673 TI - Perceived racial/ethnic discrimination among fifth-grade students and its association with mental health. AB - OBJECTIVES: We sought to describe the prevalence, characteristics, and mental health problems of children who experience perceived racial/ethnic discrimination. METHODS: We analyzed cross-sectional data from a study of 5147 fifth-grade students and their parents from public schools in 3 US metropolitan areas. We used multivariate logistic regression (overall and stratified by race/ethnicity) to examine the associations of sociodemographic factors and mental health problems with perceived racial/ethnic discrimination. RESULTS: Fifteen percent of children reported perceived racial/ethnic discrimination, with 80% reporting that discrimination occurred at school. A greater percentage of Black (20%), Hispanic (15%), and other (16%) children reported perceived racial/ethnic discrimination compared with White (7%) children. Children who reported perceived racial/ethnic discrimination were more likely to have symptoms of each of the 4 mental health conditions included in the analysis: depression, attention deficit hyperactivity disorder, oppositional defiant disorder, and conduct disorder. An association between perceived racial/ethnic discrimination and depressive symptoms was found for Black, Hispanic, and other children but not for White children. CONCLUSIONS: Perceived racial/ethnic discrimination is not an uncommon experience among fifth-grade students and may be associated with a variety of mental health disorders. PMID- 19299674 TI - Seven-year life outcomes of adolescent offenders in Los Angeles. AB - OBJECTIVES: We examined important life outcomes for adolescent offenders to describe how they were faring in young adulthood. METHODS: We assessed 449 adolescent offenders (aged 13-17 years) in Los Angeles, CA, whose cases had been adjudicated by the Los Angeles Superior Court and who had been referred to group homes between February 1999 and May 2000. We used the Global Appraisal of Individual Needs to interview respondents at baseline and at 3, 6, 12, 72, and 87 months after baseline. A total of 395 respondents (88%) were interviewed or confirmed as dead at the final interview. RESULTS: At final interview, 12 respondents had died, 7 of them from gunshot wounds. Thirty-six percent of respondents reported recent hard drug use, and 27% reported 5 or more symptoms of substance dependence. Sixty-six percent reported committing an illegal activity within the previous year, 37% reported being arrested within the previous year, and 25% reported being in jail or prison every day for the previous 90 days. Fifty-eight percent had completed high school or obtained a GED, and 63% reported working at a job in the previous year. CONCLUSIONS: The high rates of negative life outcomes presented here suggest the need for more effective rehabilitation programs for juvenile offenders. PMID- 19299675 TI - Eating when there is not enough to eat: eating behaviors and perceptions of food among food-insecure youths. AB - OBJECTIVES: We explored differences in adolescents' eating habits, perceptions, and dietary intakes by food security status. METHODS: As part of Project EAT (Eating Among Teens), we surveyed 4746 multiethnic middle and high school students in 31 primarily urban schools in the Minneapolis-St. Paul, Minnesota, area during the 1998-1999 academic year. Participants completed in-class surveys. We used multiple regression analysis to characterize associations between behaviors, perceptions, nutritional intake, and food security status. RESULTS: Compared with food-secure youths, food-insecure youths were more likely to perceive that eating healthfully was inconvenient and that healthy food did not taste good. Additionally, food-insecure youths reported eating more fast food but fewer family meals and breakfasts per week than did youths who were food secure. Food-insecure and food-secure youths perceived similar benefits from eating healthfully (P = .75). Compared with those who were food secure, food-insecure youths had higher fat intakes (P < .01). Food-insecure youths were more likely to have a body mass index above the 95th percentile. CONCLUSIONS: The eating patterns of food-insecure adolescents differ in important ways from the eating patterns of those who are food secure. Policies and interventions focusing on improving the foods that these youths eat deserve further examination. PMID- 19299676 TI - Socioeconomic inequality in exposure to bullying during adolescence: a comparative, cross-sectional, multilevel study in 35 countries. AB - OBJECTIVES: We examined the socioeconomic distribution of adolescent exposure to bullying internationally and documented the contribution of the macroeconomic environment. METHODS: We used an international survey of 162,305 students aged 11, 13, and 15 years from nationally representative samples of 5998 schools in 35 countries in Europe and North America for the 2001-2002 school year. The survey used standardized measures of exposure to bullying and socioeconomic affluence. RESULTS: Adolescents from families of low affluence reported higher prevalence of being victims of bullying (odds ratio [OR] = 1.13; 95% confidence interval [CI] = 1.10, 1.16). International differences in prevalence of exposure to bullying were not associated with the economic level of the country (as measured by gross national income) or the school, but wide disparities in affluence at a school and large economic inequality (as measured by the Gini coefficient) at the national level were associated with an increased prevalence of exposure to bullying. CONCLUSIONS: There is socioeconomic inequality in exposure to bullying among adolescents, leaving children of greater socioeconomic disadvantage at higher risk of victimization. Adolescents who attend schools and live in countries where socioeconomic differences are larger are at higher risk of being bullied. PMID- 19299677 TI - Public health workforce enumeration. AB - Comprehensive data on the public health workforce are fundamental to workforce development throughout the public health system. Such information is also a critical data element in public health systems research, a growing area of study that can inform the practice of public health at all levels. However, methodologic and institutional issues challenge the development of comparable indicators for the federal, state, and local public health workforce. A 2006-2007 Association of State and Territorial Health Officials workforce enumeration pilot project demonstrated the issues involved in collecting workforce data. This project illustrated key elements of an institutionalized national system of workforce enumeration, which would be needed for a robust, recurring count that provides a national picture of the public health workforce. PMID- 19299678 TI - Effectiveness of Cultivando la Salud: a breast and cervical cancer screening promotion program for low-income Hispanic women. AB - OBJECTIVES: We tested the effectiveness of a lay health worker intervention to increase breast and cervical cancer screening among low-income Hispanic women. METHODS: Participants were women 50 years and older who were nonadherent to mammography (n = 464) or Papanicolaou (Pap) test (n = 243) screening guidelines. After the collection of baseline data, lay health workers implemented the Cultivando la Salud (CLS; Cultivating Health) intervention. Data collectors then interviewed the participants 6 months later. RESULTS: At follow-up, screening completion was higher among women in the intervention group than in the control group for both mammography (40.8% vs 29.9%; P < .05) and Pap test (39.5% vs 23.6%; P < .05) screening. In an intent-to-treat analysis, these differences remained but were not significant. The intervention increased mammography self efficacy, perceived susceptibility, perceived survivability, perceived benefits of mammography, subjective norms, and processes of change. The intervention also significantly increased Pap test self-efficacy, perceived benefits of having a Pap test, subjective norms, and perceived survivability of cancer. It did not change Pap test knowledge, perceived susceptibility, or perceptions about negative aspects of Pap test screening. CONCLUSIONS: Our results add to the evidence concerning the effectiveness of lay health worker interventions for increasing Pap test screening and mammography. Future research should explore the effectiveness of CLS in other Hispanic groups, the mechanisms through which interpersonal communication influences decisions about screening, and how effective interventions such as CLS can best be adopted and implemented in community-based organizations or other settings. PMID- 19299679 TI - An observational study of consumers' accessing of nutrition information in chain restaurants. AB - In this observational study, we determined how frequently consumers accessed on premises nutrition information provided at chain restaurants. The number of patrons entering and accessing nutrition information was recorded at 8 locations that were part of 4 major restaurant chains (McDonald's, Burger King, Starbucks, and Au Bon Pain). Only 6 (0.1%) of 4311 patrons accessed on-premises nutrition information before purchasing food. This very small percentage suggests that such information should be more prominently displayed, such as on restaurant menu boards, to help customers make informed decisions. PMID- 19299680 TI - Hospital practices and women's likelihood of fulfilling their intention to exclusively breastfeed. AB - OBJECTIVES: We sought to assess whether breastfeeding-related hospital practices reported by mothers were associated with achievement of their intentions to exclusively breastfeed. METHODS: We used data from Listening to Mothers II, a nationally representative survey of 1573 mothers who had given birth in a hospital to a singleton in 2005. Mothers were asked retrospectively about their breastfeeding intention, infant feeding at 1 week, and 7 hospital practices. RESULTS: Primiparas reported a substantial difference between their intention to exclusively breastfeed (70%) and this practice at 1 week (50%). They also reported hospital practices that conflicted with the Baby-Friendly Ten Steps, including supplementation (49%) and pacifier use (45%). Primiparas who delivered in hospitals that practiced 6 or 7 of the steps were 6 times more likely to achieve their intention to exclusively breastfeed than were those in hospitals that practiced none or 1 of the steps. Mothers who reported supplemental feedings to their infant were less likely to achieve their intention to exclusively breastfeed: primiparas (adjusted odds ratio [AOR] = 4.4; 95% confidence interval [CI] = 2.1, 9.3); multiparas (AOR = 8.8; 95% CI = 4.4, 17.6). CONCLUSIONS: Hospitals should implement policies that support breastfeeding with particular attention to eliminating supplementation of healthy newborns. PMID- 19299681 TI - A multifaceted intervention to improve health worker adherence to integrated management of childhood illness guidelines in Benin. AB - OBJECTIVES: We evaluated an intervention to support health workers after training in Integrated Management of Childhood Illness (IMCI), a strategy that can improve outcomes for children in developing countries by encouraging workers' use of evidence-based guidelines for managing the leading causes of child mortality. METHODS: We conducted a randomized trial in Benin. We administered a survey in 1999 to assess health care quality before IMCI training. Health workers then received training plus either study supports (job aids, nonfinancial incentives, and supervision of workers and supervisors) or usual supports. Follow-up surveys conducted in 2001 to 2004 assessed recommended treatment, recommended or adequate treatment, and an index of overall guideline adherence. RESULTS: We analyzed 1244 consultations. Performance improved in both intervention and control groups, with no significant differences between groups. However, training proceeded slowly, and low-quality care from health workers without IMCI training diluted intervention effects. Per-protocol analyses revealed that workers with IMCI training plus study supports provided better care than did those with training plus usual supports (27.3 percentage-point difference for recommended treatment; P < .05), and both groups outperformed untrained workers. CONCLUSIONS: IMCI training was useful but insufficient. Relatively inexpensive supports can lead to additional improvements. PMID- 19299682 TI - The effect of aerobic training and cardiac autonomic regulation in young adults. AB - OBJECTIVES: We tested the effect of aerobic exercise on autonomic regulation of the heart in healthy young adults. METHODS: Healthy, sedentary young adults (n = 149; age = 30.4 +/- 7.53 years) were randomized to receive 12 weeks of either aerobic conditioning or strength training. Primary outcomes were heart rate and RR interval variability (RRV) measured before and after training and after 4 weeks of sedentary deconditioning. RRV, a noninvasive index of cardiac autonomic regulation, reflects variability in the intervals between consecutive R waves of the electrocardiogram. RESULTS: Aerobic conditioning but not strength training led to a significant increase in aerobic capacity (3.11 mL/kg/min), a decrease in heart rate (-3.49 beats per minute), and an increase in high-frequency RRV (0.25 natural log msec2), each of which returned to pretraining levels after deconditioning. Significant 3-way interactions, however, revealed autonomic effects only in men. CONCLUSIONS: In sedentary, healthy young adults, aerobic conditioning but not strength training enhances autonomic control of the heart, but post hoc analyses suggested that gender plays a significant role in this exercise-related cardioprotection. PMID- 19299683 TI - Association between neighborhood context and smoking prevalence among Asian Americans. AB - OBJECTIVES: To study neighborhood-level determinants of smoking among Asian Americans, we examined 3 neighborhood factors (ethnic enclave, socioeconomics, and perceived social cohesion) and smoking prevalence in a population-based sample. METHODS: We linked data from the 2003 California Health Interview Survey to tract-level data from the 2000 Census. We used multivariate logistic regression models to estimate the associations between smoking and neighborhood level factors, independent of individual factors. RESULTS: Twenty-two percent of 1693 Asian men and 6% of 2174 Asian women reported current smoking. Women living in an Asian enclave were less likely to smoke (adjusted odds ratio [AOR] = 0.27; 95% confidence interval [CI] = 0.08, 0.88). Among men, higher levels of perceived neighborhood social cohesion were associated with lower odds of smoking (AOR = 0.74; 95% CI = 0.61, 0.91). CONCLUSIONS: The association between contextual factors and smoking differed for men and women. For women, living in an Asian enclave may represent cultural behavioral norms. For men, neighborhood trust and cohesiveness may buffer stress. Smoking prevention and cessation interventions among Asian Americans may be more effective if they address contextual factors. PMID- 19299684 TI - Loneliness as a sexual risk factor for male Mexican migrant workers. AB - HIV/AIDs risk among migrant workers is often examined through individual determinants with limited consideration of social context. We used data from systematic ethnographic observations, structured interviews (n = 50), and life history interviews (n = 10) to examine the relationship between loneliness and HIV/AIDS risk for recently arrived (within the last 3 years) male Mexican migrant workers in New York City. Higher levels of loneliness were strongly associated with frequency of sexual risk behavior (r = 0.64; P = .008). From our ethnographic observations, we found that loneliness was a dominant element in workers' migration experience and that 2 different kinds of social spaces served as supportive environments for dealing with loneliness: bars or dance clubs and Catholic churches. Loneliness should be addressed as a critical factor in reducing HIV/AIDS risk among Mexican male migrant workers. PMID- 19299685 TI - "... But then he became my sipa": the implications of relationship fluidity for condom use among women sex workers in Antananarivo, Madagascar. AB - Increasing evidence indicates that sex workers use condoms less consistently with regular (i.e., nonpaying) partners than with clients. Few studies have examined the extent to which these 2 categories are mutually exclusive. In an ethnographic study of women's sex work in Antananarivo, Madagascar, we examined how the meaning of women sex workers' sexual relationships could shift among 3 different forms of sex work. Condom use was less likely in forms in which the distinction between client and lover (sipa in Malagasy) was fluid. For many sex workers, therefore, relationships they understood to be intimate imparted the greatest health vulnerability. It is important to examine the influence of the meaning of sexual relationships on condom use for HIV prevention. Policy implications for HIV prevention work with sex workers are considered. PMID- 19299686 TI - Impoverished women with children and no welfare benefits: the urgency of researching failures of the Temporary Assistance for Needy Families program. AB - In the United States, the numbers of impoverished women with children and no cash safety net are increasing and constitute an emerging population. Many have exhausted cash benefits from Temporary Assistance for Needy Families, the work based welfare program that replaced Aid to Families With Dependent Children in 1996. We examine empirical evidence about poverty and use of welfare programs in the United States, jobs for women on welfare, the consequences of leaving welfare, health disparities disproportionate to those of the general population, and outcomes for children of needy families. It is important that public health researchers investigate the experiences of the families for whom Temporary Assistance for Needy Families has failed. PMID- 19299687 TI - Comment on "the anergic state in sarcoidosis is associated with diminished dendritic cell function". PMID- 19299688 TI - The unexpected pleiotropic activities of RANTES. PMID- 19299689 TI - A human T cell-specific molecule is a member of a new gene family. 1988. PMID- 19299690 TI - Cutting Edge: Ikaros null thymocytes mature into the CD4 lineage with reduced TCR signal: A study using CD3{zeta} immunoreceptor tyrosine-based activation motif transgenic mice. AB - Positive selection is a critical T cell developmental checkpoint that is driven by TCR signals. Enhanced positive selection toward the CD4 lineage occurs in the absence of Ikaros. One explanation for this phenotype is that Ikaros establishes the TCR signaling threshold that must be overcome for positive selection to occur. In the current study, this possibility is explored through the use of CD3zeta ITAM transgenic mice that express a CD3 zeta-chain with zero, one, or three ITAMs and an MHC class II (DO11.10)- or MHC class I (H-Y)-restricted TCR transgene. Using this system, we demonstrate that in the absence of Ikaros, thymocytes are able to mature into the CD4 lineage with reduced TCR signaling potential compared with that required to drive the maturation of wild-type thymocytes. We also demonstrate that maturation into the CD8 lineage is enhanced under conditions of reduced TCR signaling potential in the absence of Ikaros. PMID- 19299691 TI - Cutting Edge: TLR-Dependent viral recognition along with type I IFN positive feedback signaling masks the requirement of viral replication for IFN-{alpha} production in plasmacytoid dendritic cells. AB - Plasmacytoid dendritic cells (pDCs) recognize RNA virus infection via TLRs and consequently produce vast amounts of type I IFN. Because nucleic acid-sensing TLRs reside in the intracellular membrane compartment, it is presumable that pDCs do not require cytoplasmic viral replication to recognize the infection. By checking Newcastle disease virus (NDV) RNA abundance in GFP(+) and GFP(-) pDCs from Ifna6gfp mice, we found that NDV replication was not detected in IFN producing pDCs. GFP(+) pDC was induced in response to replication-incompetent NDV. In contrast, the replication-incompetent NDV failed to induce IFN-producing pDCs in type I IFNR-deficient mice. The lack of IFNR signaling led to the replication of NDV and the subsequent RIG-I-like helicase-dependent IFN-alpha production in pDCs. These results showed that detection of viruses via TLRs together with a type I IFN feedback system circumvents the requirement for viral replication-dependent recognition in pDCs. PMID- 19299692 TI - Cutting Edge: A2B Adenosine receptor signaling provides potent protection during intestinal ischemia/reperfusion injury. AB - Gastrointestinal ischemia/reperfusion (IR) injury significantly contributes to the morbidity and mortality of critical illness. In this study, we hypothesized a protective role for extracellular adenosine signaling in intestinal IR injury. Initial profiling studies of mucosal scrapings following murine IR demonstrated selective induction of the A2B adenosine receptor (A2BAR) transcript. Moreover, gene-targeted mice for the A2BAR showed more profound intestinal IR injury compared with controls. In contrast, A2AAR(-/-) mice exhibited no differences in intestinal injury compared with littermate controls. In addition, selective inhibition of the A2BAR resulted in enhanced intestinal inflammation and injury during IR. Furthermore, A2BAR agonist treatment (BAY 60-6583) protected from intestinal injury, inflammation, and permeability dysfunction in wild-type mice, whereas the therapeutic effects of BAY 60-6583 were abolished following targeted A2BAR gene deletion. Taken together, these studies demonstrate the A2BAR as a novel therapeutic target for protection during gastrointestinal IR injury. PMID- 19299693 TI - Cutting edge: Acute lung allograft rejection is independent of secondary lymphoid organs. AB - It is the prevailing view that adaptive immune responses are initiated in secondary lymphoid organs. Studies using alymphoplastic mice have shown that secondary lymphoid organs are essential to initiate allograft rejection of skin, heart, and small bowel. The high immunogenicity of lungs is well recognized and allograft rejection remains a major contributing factor to poor outcomes after lung transplantation. We show in this study that alloreactive T cells are initially primed within lung allografts and not in secondary lymphoid organs following transplantation. In contrast to other organs, lungs are acutely rejected in the absence of secondary lymphoid organs. Two-photon microscopy revealed that recipient T cells cluster predominantly around lung-resident, donor derived CD11c(+) cells early after engraftment. These findings demonstrate for the first time that alloimmune responses following lung transplantation are initiated in the graft itself and therefore identify a novel, potentially clinically relevant mechanism of lung allograft rejection. PMID- 19299694 TI - Cutting edge: Dok-1 and Dok-2 adaptor molecules are regulated by phosphatidylinositol 5-phosphate production in T cells. AB - Downstream of tyrosine kinase (Dok) proteins Dok-1 and Dok-2 are involved in T cell homeostasis maintenance. Dok protein tyrosine phosphorylation plays a key role in establishing negative feedback loops of T cell signaling. These structurally related adapter molecules contain a pleckstrin homology (PH) domain generally acting as a lipid/protein-interacting module. We show that the presence of this PH domain is necessary for the tyrosine phosphorylation of Dok proteins and their negative functions in T cells. We find that Dok-1/Dok-2 PH domains bind in vitro to the rare phosphoinositide species, phosphatidylinositol 5-phosphate (PtdIns5P). Dok tyrosine phosphorylation correlates with PtdIns5P production in T cells upon TCR triggering. Furthermore, we demonstrate that PtdIns5P increase regulates Dok tyrosine phosphorylation in vivo. Together, our data identify a novel lipid mediator in T cell signaling and suggest that PH-PtdIns5P interactions regulate T cell responses. PMID- 19299695 TI - CD4-positive T lymphocytes provide a neuroimmunological link in the control of adult hippocampal neurogenesis. AB - Adult hippocampal neurogenesis occurs in an exceptional permissive microenvironment. Neuroimmunological mechanisms might be prominently involved in the endogenous homeostatic principles that control baseline levels of adult neurogenesis. We show in this study that this homeostasis is partially dependent on CD4-positive T lymphocytes. Systemic depletion of CD4-positive T lymphocytes led to significantly reduced hippocampal neurogenesis, impaired reversal learning in the Morris water maze, and decreased brain-derived neurotrophic factor expression in the brain. No such effect of CD8 or B cells was observed. Repopulation of RAG2(-/-) mice with CD4, but not with CD8 cells again increased precursor cell proliferation. The T cells in our experiments were non-CNS specific and rarely detectable in the healthy brain. Thus, we can exclude cell cell contacts between immune and brain cells or lymphocyte infiltration into the CNS as a prerequisite for an effect of CD4-T cells on neurogenesis. We propose that systemic CD4-T cell activity is required for maintaining cellular plasticity in the adult hippocampus and represents an evolutionary relevant communication route for the brain to respond to environmental changes. PMID- 19299696 TI - Anti-CD25 antibody-mediated depletion of effector T cell populations enhances susceptibility of mice to acute but not chronic Toxoplasma gondii infection. AB - Natural regulatory T cells (Tregs) constitutively express the IL-2R alpha-chain (CD25) on their surface. Consequently, administration of anti-CD25 Abs is a commonly used technique to deplete Treg populations in vivo. However, activated effector T cells may also transiently express CD25, and are thus also potential targets for anti-CD25 Abs. In this study using Toxoplasma gondii as a model proinflammatory infection, we have examined the capacity of anti-CD25 Abs to target effector T cell populations during an inflammatory episode, to determine to what extent that this action may modulate the outcome of disease. Anti-CD25 Ab treated C57BL/6 mice displayed significantly reduced CD4(+) T cell IFN-gamma production during acute T. gondii infection and exhibited reduced weight loss and liver pathology during early acute infection; aspects of infection previously associated with effector CD4(+) T cell responses. In agreement, anti-CD25 Ab administration impaired parasite control and caused mice to succumb to infection during late acute/early chronic stages of infection with elevated tissue parasite burdens. In contrast, anti-CD25 Ab treatment of mice with established chronic infections did not markedly affect brain parasite burdens, suggesting that protective T cell populations do not express CD25 during chronic stages of T. gondii infection. In summary, we have demonstrated that anti-CD25 Abs may directly abrogate effector T cell responses during an inflammatory episode, highlighting important limitations of the use of anti-CD25 Ab administration to examine Treg function during inflammatory settings. PMID- 19299697 TI - Protective antigen-independent CD8 T cell memory is maintained during {gamma} herpesvirus persistence. AB - Ag persistence during high-titer chronic viral infections induces CD8 T cell dysfunction and lack of Ag-independent CD8 T cell memory formation. However, we have a poor understanding of the generation and maintenance of CD8 T cell memory during asymptomatic persistent viral infections, particularly gamma-herpesvirus infections. In this study, we demonstrate that the continuous presence of cognate Ag in the host is not required for the maintenance of CD8 T cell memory during a persistent gamma-herpesvirus infection. Importantly, the Ag-independent CD8 T cell memory that is maintained during gamma-herpesvirus persistence has the capacity to survive long-term under homeostatic conditions and to mount a protective recall response to a secondary encounter with the pathogen. These data highlight the ability of the immune system to maintain a population of protective memory CD8 T cells with capacity for long-term Ag-independent survival in the presence of systemic virus persistence. PMID- 19299698 TI - Recombinant Ov-ASP-1, a Th1-biased protein adjuvant derived from the helminth Onchocerca volvulus, can directly bind and activate antigen-presenting cells. AB - We previously reported that rOv-ASP-1, a recombinant Onchocerca volvulus activation associated protein-1, was a potent adjuvant for recombinant protein or synthetic peptide-based Ags. In this study, we further evaluated the adjuvanticity of rOv-ASP-1 and explored its mechanism of action. Consistently, recombinant full-length spike protein of SARS-CoV or its receptor-binding domain in the presence of rOv-ASP-1 could effectively induce a mixed but Th1-skewed immune response in immunized mice. It appears that rOv-ASP-1 primarily bound to the APCs among human PBMCs and triggered Th1-biased proinflammatory cytokine production probably via the activation of monocyte-derived dendritic cells and the TLR, TLR2, and TLR4, thus suggesting that rOv-ASP-1 is a novel potent innate adjuvant. PMID- 19299699 TI - Inhibition of cAMP degradation improves regulatory T cell-mediated suppression. AB - Naturally occurring regulatory T cells (nTreg cells) are crucial for the maintenance of peripheral tolerance. We have previously shown that a key mechanism of their suppressive action is based on a contact-dependent transfer of cAMP from nTreg cells to responder T cells. Herein, we further elucidate the important role of cAMP for the suppressive properties of nTreg cells. Prevention of cAMP degradation by application of the phosphodiesterase 4 inhibitor rolipram led to strongly increased suppressive potency of nTreg cells for Th2 cells in vitro and in vivo. Detailed analyses revealed that rolipram caused, in the presence of nTreg cells, a synergistic increase of cAMP in responder Th2 cells. In vivo, the application of nTreg cells in a strictly Th2-dependent preclinical model of asthma had only a marginal effect. However, the additional treatment with rolipram led to a considerable reduction of airway hyperresponsiveness and inflammation in a prophylactic as well as in a therapeutic model. This amelioration was correlated with enhanced cAMP-levels in lung Th2 cells in vivo. Collectively, these data support our observation that cAMP has a key function for nTreg cell-based suppression and they clearly demonstrate that the effect of cAMP on T responder cells can be greatly enhanced upon application of phosphodiesterase 4 inhibitors. PMID- 19299700 TI - Histoplasma capsulatum cell wall {beta}-glucan induces lipid body formation through CD18, TLR2, and dectin-1 receptors: correlation with leukotriene B4 generation and role in HIV-1 infection. AB - Histoplasma capsulatum (Hc) is a facultative, intracellular parasite of worldwide significance. Infection with Hc produces a broad spectrum of diseases and may progress to a life-threatening systemic disease, particularly in individuals with HIV infection. Resolution of histoplasmosis is associated with the activation of cell-mediated immunity, and leukotriene B(4) plays an important role in this event. Lipid bodies (LBs) are increasingly being recognized as multifunctional organelles with roles in inflammation and infection. In this study, we investigated LB formation in histoplasmosis and its putative function in innate immunity. LB formation in leukocytes harvested from Hc-infected C57BL/6 mice peaks on day 2 postinfection and correlates with enhanced generation of lipid mediators, including leukotriene B(4) and PGE(2). Pretreatment of leukocytes with platelet-activating factor and BLT1 receptor antagonists showed that both lipid mediators are involved in cell signaling for LB formation. Alveolar leukocytes cultured with live or dead Hc also presented an increase in LB numbers. The yeast alkali-insoluble fraction 1, which contains mainly beta-glucan isolated from the Hc cell wall, induced a dose- and time-dependent increase in LB numbers, indicating that beta-glucan plays a signaling role in LB formation. In agreement with this hypothesis, beta-glucan-elicited LB formation was inhibited in leukocytes from 5-LO(-/-), CD18(low) and TLR2(-/-) mice, as well as in leukocytes pretreated with anti-Dectin-1 Ab. Interestingly, human monocytes from HIV-1 infected patients failed to produce LBs after beta-glucan stimulation. These results demonstrate that Hc induces LB formation, an event correlated with eicosanoid production, and suggest a role for these lipid-enriched organelles in host defense during fungal infection. PMID- 19299701 TI - Cross-linking of GM1 ganglioside by galectin-1 mediates regulatory T cell activity involving TRPC5 channel activation: possible role in suppressing experimental autoimmune encephalomyelitis. AB - Several animal autoimmune disorders are suppressed by treatment with the GM1 cross-linking units of certain toxins such as B subunit of cholera toxin (CtxB). Due to the recent observation of GM1 being a binding partner for the endogenous lectin galectin-1 (Gal-1), which is known to ameliorate symptoms in certain animal models of autoimmune disorders, we tested the hypothesis that an operative Gal-1/GM1 interplay induces immunosuppression in a manner evidenced by both in vivo and in vitro systems. Our study of murine experimental autoimmune encephalomyelitis (EAE) indicated suppressive effects by both CtxB and Gal-1 and further highlighted the role of GM1 in demonstrating enhanced susceptibility to EAE in mice lacking this ganglioside. At the in vitro level, polyclonal activation of murine regulatory T (Treg) cells caused up-regulation of Gal-1 that was both cell bound and released to the medium. Similar activation of murine CD4(+) and CD8(+) effector T (Teff) cells resulted in significant elevation of GM1 and GD1a, the neuraminidase-reactive precursor to GM1. Activation of Teff cells also up-regulated TRPC5 channels which mediated Ca(2+) influx upon GM1 cross-linking by Gal-1 or CtxB. This involved co-cross-linking of heterodimeric integrin due to close association of these alpha(4)beta(1) and alpha(5)beta(1) glycoproteins with GM1. Short hairpin RNA (shRNA) knockdown of TRPC5 in Teff cells blocked contact-dependent proliferation inhibition by Treg cells as well as Gal-1/CtxB-triggered Ca(2+) influx. Our results thus indicate GM1 in Teff cells to be the primary target of Gal-1 expressed by Treg cells, the resulting co-cross linking and TRPC5 channel activation contributing importantly to the mechanism of autoimmune suppression. PMID- 19299702 TI - Autophagy is essential for mitochondrial clearance in mature T lymphocytes. AB - Macroautophagy plays an important role in the regulation of cell survival, metabolism, and the lysosomal degradation of cytoplasmic material. In the immune system, autophagy contributes to the clearance of intracellular pathogens, MHCII cross-presentation of endogenous Ags, as well as cell survival. We and others have demonstrated that autophagy occurs in T lymphocytes and contributes to the regulation of their cellular function, including survival and proliferation. Here we show that the essential autophagy gene Atg7 is required in a cell-intrinsic manner for the survival of mature primary T lymphocytes. We also find that mitochondrial content is developmentally regulated in T but not in B cells, with exit from the thymus marking a transition from high mitochondrial content in thymocytes to lower mitochondrial content in mature T cells. Macroautophagy has been proposed to play an important role in the clearance of intracellular organelles, and autophagy-deficient mature T cells fail to reduce their mitochondrial content in vivo. Consistent with alterations in mitochondrial content, autophagy-deficient T cells have increased reactive oxygen species production as well as an imbalance in pro- and antiapoptotic protein expression. With much recent interest in the possibility of autophagy-dependent developmentally programmed clearance of organelles in lens epithelial cells and erythrocytes, our data demonstrate that autophagy may have a physiologically significant role in the clearance of superfluous mitochondria in T lymphocytes as part of normal T cell homeostasis. PMID- 19299703 TI - Human benign prostatic hyperplasia stromal cells as inducers and targets of chronic immuno-mediated inflammation. AB - Benign prostatic hyperplasia (BPH), a highly prevalent prostatic condition, could involve an inflammatory component in disease pathogenesis. In this study, we show that human stromal prostate cells obtained from BPH tissue can actively contribute to the inflammatory process by secreting proinflammatory cytokines as well as chemokines able to recruit lymphomonuclear cells and by acting as APCs. BPH cells express all of the TLRs and their ligation leads to the secretion of CXCL8/IL-8, CXCL10, and IL-6. In addition, BPH cells express costimulatory as well as class I and class II MHC molecules, which activate alloreactive CD4(+) cells that in turn markedly up-regulate IL-12/IL-23p40 and IL-12p75 secretion by BPH cells. Alloreactive CD4(+) cells activated by BPH cells secrete IFN-gamma and IL-17. These cytokines up-regulate IL-6, IL-8, and CXCL10 production by BPH cells, creating a positive feedback loop that can amplify inflammation. IL-8 induces autocrine/paracrine proliferation of BPH cells, indicating also a growth promoting activity of this chemokine in disease pathogenesis. These results show that human BPH cells represent nonprofessional APCs able to induce and sustain chronic inflammatory processes, supporting the relevance of inflammation in BPH pathogenesis. PMID- 19299704 TI - Reduced c-myc expression levels limit follicular mature B cell cycling in response to TLR signals. AB - The splenic B cell compartment is comprised of two major, functionally distinct, mature B cell subsets, i.e., follicular mature (FM) and marginal zone (MZ) B cells. Whereas MZ B cells exhibit a robust proliferative response following stimulation with the TLR4 ligand LPS, FM B cells display markedly delayed and reduced levels of proliferation to the identical stimulus. The current study was designed to identify a potential mechanism(s) accounting for this differential responsiveness. In contrast to the delay in cell cycle entry, FM and MZ B cells exhibited nearly identical LPS-driven alterations in the expression level of cell surface activation markers. Furthermore, both the NF-kappaB and mTOR signaling cascades were similarly activated by LPS stimulation in FM vs MZ B cells, while inducible activation of ERK and AKT were nearly absent in both subsets. MZ B cells, however, exhibited higher basal levels of phospho-AKT and pS6, consistent with a preactivated status. Importantly, both basal and LPS activation-induced c myc expression was markedly reduced in FM vs MZ B cells and enforced c-myc expression fully restored the defective proliferative response in FM B cells. These data support a model wherein TLR responses in FM B cells are tightly regulated by limiting c-myc levels, thereby providing an important checkpoint to control nonspecific FM B cell activation in the absence of cognate Ag. PMID- 19299705 TI - ICOS controls effector function but not trafficking receptor expression of kidney infiltrating effector T cells in murine lupus. AB - Renal pathology in systemic lupus erythematosus involves both autoantibody deposition and a cellular inflammatory response, both of which are mediated by effector CD4 T cells. MRL(lpr) mice spontaneously develop massive perivascular infiltrates, but the pathways that regulate the development, trafficking, and effector functions of kidney-infiltrating T cells are poorly defined. To address these questions, we first surveyed inflammatory chemokine protein levels in nephritic kidneys from lupus-prone MRL(lpr) mice. After identifying highly elevated levels of the CXCR3 ligand CXCL9, we found that kidney-infiltrating effectors are enriched for expression of CXCR3, as well as P-selectin ligand and ICOS. Using genetic ablation, we demonstrate that ICOS plays an essential role in the establishment of renal perivascular infiltrates, although a small number of infiltrating cells remain around the blood vessels. Interestingly, though IgG autoantibody production is substantially reduced in Icos(-/-) MRL(lpr) mice, the progression of immune complex glomerulonephritis is only modestly diminished and the production of inflammatory chemokines, such as CXCL9, remains high in the kidney. We find that Icos(-/-) effector cell numbers are only slightly reduced and these have normal expression of CXCR3 and P-selectin ligand with intact migration to CXCL9. However, they have impaired production of inflammatory cytokines and fail to show evidence of efficient proliferation in the kidney. Thus, while dispensable for acquisition of renal trafficking receptor expression, ICOS is strictly required for local inflammatory functions of autoreactive CD4 T cells in murine lupus. PMID- 19299706 TI - Vascular endothelial growth factor C facilitates immune tolerance and endovascular activity of human uterine NK cells at the maternal-fetal interface. AB - Although replete with cytotoxic machinery, uterine NK (uNK) cells remain tolerant at the maternal-fetal interface. The mechanisms that facilitate the uNK cell tolerance are largely unknown. In this study, we demonstrate that vascular endothelial growth factor (VEGF) C, a proangiogenic factor produced by uNK cells, is responsible for their noncytotoxic activity. VEGF C-producing uNK cells support endovascular processes as demonstrated in a three-dimensional coculture model of capillary tube formation on Matrigel. Peripheral blood NK cells fail to produce VEGF C and remain cytotoxic. This response can be reversed by exogenous VEGF C. We show that cytoprotection by VEGF C can be related to induction of the TAP-1 expression and MHC class I assembly in target cells. Small interfering RNA mediated silencing of TAP-1 expression abolished the VEGF C-imparted protection. Overall, these results demonstrate that empowerment of uNK cells with angiogenic factors keeps them noncytotoxic. This phenotype is critical to their pregnancy compatible immunovascular role during placentation and fetal development. PMID- 19299707 TI - PTPN22 deficiency cooperates with the CD45 E613R allele to break tolerance on a non-autoimmune background. AB - Pep and CD45 are tyrosine phosphatases whose targets include the Src-family kinases, critical mediators of Ag receptor signaling. A polymorphism in PTPN22, the gene that encodes the human Pep orthologue Lyp, confers susceptibility to multiple human autoimmune diseases in the context of complex genetic backgrounds. However, the functional significance of the R620W risk allele is not clear. We report that misexpression of wild-type or R620W Pep/Lyp in Jurkat cells, in the context of its binding partner Csk, unmasks the risk allele as a hypomorph. It has been shown previously that although Pep-deficient mice on the B6 background have hyperresponsive memory T cells, autoimmunity does not develop. Mice containing a point mutation in the CD45 juxtamembrane wedge domain (E613R) develop a B cell-driven, lupus-like disease on the mixed 129/B6 background, but not on the B6 background. We studied the ability of Pep deficiency to act as a genetic modifier of the CD45 E613R mutation on the nonautoimmune B6 background to understand how complex susceptibility loci might interact in autoimmunity. In this study we report that double mutant mice develop a lupus-like disease as well as lymphadenopathy, polyclonal lymphocyte activation, and accelerated memory T cell formation. Following Ag receptor stimulation, peripheral B cells in the double mutant mice phenocopy hyperresponsive CD45 E613R B cells, whereas peripheral T cells respond like Pep(-/-) T cells. These studies suggest that Pep( /-) T cells in the context of a susceptible microenvironment can drive hyperresponsive CD45 E613R B cells to break tolerance. PMID- 19299708 TI - 4-1BB functions as a survival factor in dendritic cells. AB - 4-1BB (CD137) is expressed on dendritic cells (DCs) and its biological function has remained largely unresolved. By comparing 4-1BB-intact (4-1BB(+/+)) and 4-1BB deficient (4-1BB(-/-)) DCs, we found that 4-1BB was strongly induced on DCs during the maturation and that DC maturation was normal in the absence of 4-1BB. However, DC survival rate was low in the absence of 4-1BB, which was due to the decreased Bcl-2 and Bcl-x(L) in 4-1BB(-/-) DCs compared with 4-1BB(+/+) DCs after DC maturation. Consistent with these results, 4-1BB(-/-) DCs showed an increased turnover rate in steady state and more severely decreased in spleen by injecting LPS compared with 4-1BB(+/+) DCs. When OVA-pulsed DCs were adoptively transferred to recipient mice along with OVA-specific CD4(+) T cells, 4-1BB(-/-) DCs did not properly migrate to the T cell zone in lymph nodes and poorly induced proliferation of CD4(+) T cells, although both DCs comparably expressed functional CCR7. Eventually, 4-1BB(-/-) DCs generated a reduced number of OVA specific memory CD4(+) T cells compared with 4-1BB(+/+) DCs. To further assess the role of 4-1BB on DC longevity in vivo, 4-1BB(+/+) and 4-1BB(-/-) C57BL/6 were administrated with Propionibacterium acnes that develop liver granuloma by recruiting DCs. Number and size of granuloma were reduced in the absence of 4 1BB, but the inflammatory cytokine level was comparable between the mice, which implied that the granuloma might be reduced due to the decreased longevity of DCs. These results demonstrate that 4-1BB on DCs controls the duration, DC-T interaction, and, therefore, immunogenicity. PMID- 19299709 TI - Identification and characterization of a human CD5+ pre-naive B cell population. AB - We have identified a distinct pre-naive B cell population circulating in human peripheral blood that exhibits an intermediate phenotype between transitional and naive B cells. Like human transitional B cells, these cells express CD5 but have intermediate densities of CD38, CD10, CD9, and the ABCB1 transporter compared with transitional and naive B cells. These pre-naive B cells account for a majority of circulating human CD5(+) B cells. Importantly, CD5(+) pre-naive B cells could be induced to differentiate into cells with a naive phenotype in vitro. CD5(+) pre-naive B cells show only partial responses to BCR stimulation and CD40 ligation and undergo more spontaneous apoptosis and cell death than do naive B cells, whereas BAFF/BLyS (B cell-activating factor belonging to the TNF family) did not enhance their survival compared with naive B cells. In contrast, CD5(+) pre-naive B cells carry out certain functions comparable to naive B cells, including the capacity to differentiate into plasma cells and the ability to function as APCs. Notably, an increased proportion of CD5(+) pre-naive B cells were found in peripheral blood of patients with systemic lupus erythematosus. These results have identified a unique intermediate in human naive B cell development within the peripheral blood and derangements of its homeostasis in patients with systemic lupus erythematosus. PMID- 19299710 TI - Novel subset of CD8{alpha}+ dendritic cells localized in the marginal zone is responsible for tolerance to cell-associated antigens. AB - Apoptotic cell clearance by dendritic cells (DCs) plays a crucial role in the maintenance of self-tolerance. In spleen, CD8alpha(+) DCs are thought to be responsible for this phenomenon by phagocytosing circulating apoptotic cells. However, as CD8alpha(+) DCs are believed to be predominantly localized in the T cell zone, it remains unclear how these DCs phagocytose blood-borne apoptotic cells accumulated in the marginal zone (MZ). In this study, we identified a subpopulation of CD8alpha(+) DCs responsible for tolerance induction to cell associated Ags. Among splenic CD8alpha(+) DCs, the CD103(+),CD207(+) subset was preferentially localized in the MZ and dominantly phagocytosed blood-borne apoptotic cells. After phagocytosis of apoptotic cells, this DC subset migrated into the T cell zone for cross-presentation of cell-associated Ags. Stimulation of TLRs induced the disappearance of this DC subset. Consequently, CD8alpha(+) DCs neither phagocytosed injected apoptotic cells nor presented cell-associated Ags in mice treated with TLR ligands. Transient ablation of this DC subset by cytochrome c injection resulted in a failure of tolerance induction to cell associated Ags, indicating that this DC subset is essential for tolerance induction by apoptotic cell clearance. PMID- 19299711 TI - Nitrated {alpha}-synuclein-induced alterations in microglial immunity are regulated by CD4+ T cell subsets. AB - Microglial inflammatory neuroregulatory activities affect the tempo of nigrostriatal degeneration during Parkinson's disease (PD). Such activities are induced, in part, by misfolded, nitrated alpha-synuclein (N-alpha-syn) within Lewy bodies released from dying or dead dopaminergic neurons. Such pathobiological events initiate innate and adaptive immune responses affecting neurodegeneration. We posit that the neurobiological activities of activated microglia are affected by cell-protein and cell-cell contacts, in that microglial interactions with N-alpha-syn and CD4(+) T cells substantively alter the microglial proteome. This leads to alterations in cell homeostatic functions and disease. CD4(+)CD25(+) regulatory T cells suppress N-alpha-syn microglial-induced reactive oxygen species and NF-kappaB activation by modulating redox-active enzymes, cell migration, phagocytosis, and bioenergetic protein expression and cell function. In contrast, CD4(+)CD25(-) effector T cells exacerbate microglial inflammation and induce putative neurotoxic responses. These data support the importance of adaptive immunity in the regulation of Parkinson's disease associated microglial inflammation. PMID- 19299712 TI - CLEC-2 is a phagocytic activation receptor expressed on murine peripheral blood neutrophils. AB - CLEC-2 is a member of the "dectin-1 cluster" of C-type lectin-like receptors and was originally thought to be restricted to platelets. In this study, we demonstrate that murine CLEC-2 is also expressed by peripheral blood neutrophils, but only weakly by bone marrow or elicited inflammatory neutrophils. On circulating neutrophils, CLEC-2 can mediate phagocytosis of Ab-coated beads and the production of proinflammatory cytokines, including TNF-alpha, in response to the CLEC-2 ligand, rhodocytin. CLEC-2 possesses a tyrosine-based cytoplasmic motif similar to that of dectin-1, and we show using chimeric analyses that the activities of this receptor are dependent on this tyrosine. Like dectin-1, CLEC-2 can recruit the signaling kinase Syk in myeloid cells, however, stimulation of this pathway does not induce the respiratory burst. These data therefore demonstrate that CLEC-2 expression is not restricted to platelets and that it functions as an activation receptor on neutrophils. PMID- 19299713 TI - Intestinal T cell responses to gluten peptides are largely heterogeneous: implications for a peptide-based therapy in celiac disease. AB - The identification of gluten peptides eliciting intestinal T cell responses is crucial for the design of a peptide-based immunotherapy in celiac disease (CD). To date, several gluten peptides have been identified to be active in CD. In the present study, we investigated the recognition profile of gluten immunogenic peptides in adult HLA-DQ2(+) celiac patients. Polyclonal, gliadin-reactive T cell lines were generated from jejunal mucosa and assayed for both proliferation and IFN-gamma production in response to 21 peptides from wheat glutenins and alpha-, gamma-, and omega-gliadins. A magnitude analysis of the IFN-gamma responses was performed to assess the hierarchy of peptide potency. Remarkably, 12 of the 14 patients recognized a different array of peptides. All alpha-gliadin stimulatory peptides mapped the 57-89 N-terminal region, thus confirming the relevance of the known polyepitope 33-mer, although it was recognized by only 50% of the patients. By contrast, gamma-gliadin peptides were collectively recognized by the great majority (11 of 14, 78%) of CD volunteers. A 17-mer variant of 33-mer, QLQPFPQPQLPYPQPQP, containing only one copy of DQ2-alpha-I and DQ2-alpha-II epitopes, was as potent as 33-mer in stimulating intestinal T cell responses. A peptide from omega-gliadin, QPQQPFPQPQQPFPWQP, although structurally related to the alpha-gliadin 17-mer, is a distinct epitope and was active in 5 out of 14 patients. In conclusion, these results showed that there is a substantial heterogeneity in intestinal T cell responses to gluten and highlighted the relevance of gamma- and omega-gliadin peptides for CD pathogenesis. Our findings indicated that alpha-gliadin (57-73), gamma-gliadin (139-153), and omega-gliadin (102-118) are the most active gluten peptides in DQ2(+) celiac patients. PMID- 19299714 TI - The cyclic AMP response element modulator {alpha} suppresses CD86 expression and APC function. AB - The cAMP response element modulator (CREM)alpha is a widely expressed transcriptional repressor that is important for the termination of the T cell immune response and contributes to the abnormal T cell function in patients with systemic lupus erythematosus. We present evidence that APCs of Crem(-/-) mice express increased amounts of the costimulatory molecule CD86 and induce enhanced Ag-dependent and Ag-independent T cell proliferation. Similarly, human APCs in which CREMalpha was selectively suppressed expressed more CD86 on the surface membrane. CREMalpha was found to bind to the CD86 promoter and suppressed its activity. Transfer of APCs from Crem(-/-) mice into naive mice facilitated a significantly stronger contact dermatitis response compared with mice into which APCs from Crem(+/+) mice had been transferred. We conclude that CREMalpha is an important negative regulator of costimulation and APC-dependent T cell function both in vitro and in vivo. PMID- 19299715 TI - Functional killer Ig-like receptors on human memory CD4+ T cells specific for cytomegalovirus. AB - Although very few CD4(+) T cells express killer Ig receptors (KIR), a large proportion of CD4(+) T cells with a late memory phenotype, characterized by the absence of CD28, does express KIR. Here, we show that KIR expression on CD4(+) T cells is also associated with memory T cell function, by showing that the frequency of CMV-specific cells is higher in CD4(+)KIR(+) than CD4(+)KIR(-) T cells. In addition, engagement of an inhibitory KIR inhibited the CMV-specific proliferation of these CD4(+)KIR(+) memory T cells, but had no detectable effect on cytokine production. Our data reveal that, in marked contrast with CD8(+) T cells, the activity of a subset of CMV-specific CD4(+) T cells is modulated by HLA class I-specific KIR. Thus, the CMV-induced down-regulation of HLA class I may in fact enhance memory CMV-specific CD4(+) T cell responses restricted by HLA class II. PMID- 19299716 TI - Ncf1 provides a reactive oxygen species-independent negative feedback regulation of TLR9-induced IL-12p70 in murine dendritic cells. AB - Permanent exposure to pathogens requires decisions toward tolerance or immunity as a prime task of dendritic cells. The molecular mechanisms preventing uncontrolled immune responses are not completely clear. We investigated the regulatory function of Ncf1, an organizing protein of NADPH oxidase, in the signaling cascade of Toll-like receptors. TLR9-stimulated spleen cells from both Ncf1-deficient and B10.Q mice with a point mutation in exon 8 of Ncf1 exhibited increased IL-12p70 secretion compared with controls. This finding was restricted to stimulatory CpG2216 and not induced by CpG2088. Because only CpG/TLR9-induced IL-12p70 was regulated by Ncf1, we used TRIF(-/-) and MyD88(-/-) cells to show that TLR9/MyD88 was primarily affected. Interestingly, additional experiments revealed that spleen cells from NOX2/gp91(phox)-deficient mice and the blocking of electron transfer by diphenylene iodonium had no influence on CpG-induced IL 12p70, confirming an NADPH oxidase-independent function of Ncf1. Finally, proving the in vivo relevance CpG adjuvant-guided OVA immunization resulted in a strong augmentation of IL-12p70-dependent Th1 IFN-gamma response only in Ncf1-deficient mice. These data suggest for the first time an important role for Ncf1 in the fine tuning of the TLR9/MyD88 pathway in vitro and in vivo that is independent of its role as an activator of NOX2. PMID- 19299717 TI - Signal transduction inhibition of APCs diminishes th17 and Th1 responses in experimental autoimmune encephalomyelitis. AB - IL-17- and IFN-gamma-secreting T cells play an important role in autoimmune responses in multiple sclerosis and the model system experimental autoimmune encephalomyelitis (EAE). Dendritic cells (DCs) in the periphery and microglia in the CNS are responsible for cytokine polarization and expansion of this T cell subset. Our results indicate that in vivo administration of a signal transduction inhibitor that targets DCs to mice with EAE led to a decrease in CNS infiltration of pathogenic Ag-specific T cells. Since this approach does not target T cells directly, we assessed the effects on the APCs that are involved in generating the T cell responses. Since in EAE and multiple sclerosis, both microglia and peripheral DCs are likely to contribute to disease, we utilized a bone marrow chimera system to distinguish between these two populations. These studies show that peripheral DCs are the primary target but that microglia are also modestly affected by CEP-701, as numbers and activation states of the cells in the CNS are decreased after therapy. Our results also showed a decrease in secretion of TNF alpha, IL-6, and IL-23 by DCs as well as a decrease in expression of costimulatory molecules. We further determined that levels of phospho-Stat1, Stat3, Stat5, and NF-kappaB, which are signaling molecules that have been implicated in these pathways, were decreased. Thus, use of this class of signal transduction inhibitors may represent a novel method to treat autoimmunity by dampening the autoreactive polarizing condition driven by DCs. PMID- 19299718 TI - Characterization of an immediate splenic precursor of CD8+ dendritic cells capable of inducing antiviral T cell responses. AB - Mouse spleens contain three major dendritic cell (DC) populations: plasmacytoid DC, conventional CD8(+)CD24(+) DC (CD8(+) DC), and conventional CD8(-)CD24(-) DC (CD8(-) DC). We have previously shown that CD8(+) DC are the major cross presenting subtype in vivo and are the main inducers of antiviral cytotoxic T lymphocyte responses. Here we show that after depletion of CD8(+) DC, the only DC capable of viral Ag presentation was a small subset that expresses CD24 but not CD8. This CD8(-)CD24(+) DC population is greatly expanded in mice treated with the DC growth factor FMS-like tyrosine kinase 3 ligand. The CD8(-)CD24(+) DC represent an immediate precursor of CD8(+) DC, as demonstrated by their expression pattern of characteristic markers of CD8(+) DC, their capacity to cross-present in vitro, and their conversion into CD8(+) DC upon adoptive transfer into recipient mice. Accordingly, the lifespan of transferred CD8( )CD24(+) DC in vivo was greatly enhanced as compared with terminally differentiated CD8(+) DC. Moreover, in a vaccination protocol, CD8(-)CD24(+) DC induced stronger T cell responses and accelerated viral clearance of HSV-1 compared with CD8(+) DC. Our results demonstrate that the ability to cross present first appears in an immediate precursor population of CD8(+) DC that does not yet express CD8. The enhanced capacity of CD8(-)CD24(+) DC to induce immune responses upon adoptive transfer makes them an attractive novel tool for DC-based immunotherapies. PMID- 19299719 TI - TLR2/MyD88-dependent and -independent activation of mast cell IgE responses by the skin commensal yeast Malassezia sympodialis. AB - Atopic eczema (AE) is a chronic inflammatory skin disease. Approximately 50% of adult AE patients have allergen-specific IgE reactivity to the skin commensal yeast Malassezia spp. Due to the ruptured skin barrier in AE, it is likely that Malassezia can come into contact with mast cells, which are known to be involved in AE. We therefore hypothesized that Malassezia spp. can activate mast cells. Bone marrow-derived mast cells (BMMCs) were generated from wild type, TLR2, TLR4, and MyD88 gene-deleted mice and cocultured with Malassezia sympodialis extract. We recorded that M. sympodialis induced release of cysteinyl leukotrienes in a dose-dependent manner in nonsensitized and IgE-anti-trinitrophenyl-sensitized BMMCs, respectively, with three times higher levels in the latter type of cells. IgE-sensitized BMMCs also responded by degranulation as assessed by release of beta-hexosaminidase, increased MCP-1 production through a MyD88-independent pathway, and activated phosphorylation of the MAPK ERK1/2. Furthermore, M. sympodialis enhanced the degranulation of IgE receptor cross-linked wild-type BMMCs and altered the IL-6 release dose-dependently. This degranulation was independent of TLR2, TLR4, and MyD88, whereas the IL-6 production was dependent on the TLR2/MyD88 pathway and MAPK signaling. In conclusion, M. sympodialis extract can activate nonsensitized and IgE-sensitized mast cells to release inflammatory mediators, to enhance the IgE-mediated degranulation of mast cells, and to modulate MAPK activation and by signaling through the TLR2/MyD88 pathway to modify the IL-6 production of IgE receptor cross-linked mast cells. Collectively, these findings indicate that M. sympodialis can activate mast cells and might thus exacerbate the inflammatory response in AE. PMID- 19299720 TI - Central role of tumor-associated CD8+ T effector/memory cells in restoring systemic antitumor immunity. AB - Sustained delivery of IL-12 and GM-CSF to tumors induces the activation of tumor resident CD8(+) T effector/memory cells (Tem) followed by cytotoxic CD8(+) T effector cell expansion. To determine whether the secondary effectors expanded from tumor-associated Tem or were primed de novo, activation kinetics of tumor draining lymph node (TDLN) CD8(+) T cells were analyzed. Treatment promoted a 4 fold increase in the numbers of TDLN CD8(+) T cells displaying a CD69(+)CCR5(+)CD62L(-) periphery-homing effector phenotype by day 4 posttherapy. Pulse labeling of tumor and TDLN T cells with BrdU confirmed that proliferation occurred exclusively within the draining lymph nodes between days 1 and 4 with subsequent migration of primed CD8(+) T effectors to tumors on day 7. Day 4 CD8(+) T effector cells preferentially homed to and lysed experimental, but not control, tumors, establishing tumor specificity. To determine whether the secondary CD8(+) T effector cell response was dependent on activation of tumor resident CD8(+) Tem, mice that were selectively depleted of tumor-infiltrating CD8(+) T cells were treated and monitored for T effector priming. In the absence of tumor-resident CD8(+) Tem, T effector cell expansion was completely abrogated in the TDLN, revealing that restoration of CD8(+) Tem function was critical to the induction of secondary T effectors. T cell priming failed to occur in IFN gamma or perforin knockout mice, demonstrating that the requirement for Tem activation was associated with induction of Tem cytotoxicity. These data confirm that intratumoral IL-12 plus GM-CSF induces de novo priming of tumor-specific CD8(+) T effector cells in the TDLN and establish the critical role of preexisting intratumoral CD8(+) Tem in driving this process. PMID- 19299721 TI - B cell proliferation, somatic hypermutation, class switch recombination, and autoantibody production in ectopic lymphoid tissue in murine lupus. AB - Intraperitoneal exposure of nonautoimmune mice to 2,6,10,14 tetramethylpentadecane (TMPD) causes lupus and the formation of ectopic lymphoid tissue. Although associated with humoral autoimmunity, it is not known whether Ab responses develop within ectopic lymphoid tissue or if B cells only secondarily migrate there. We show that ectopic lymphoid tissue induced by TMPD not only resembles secondary lymphoid tissue morphologically, but it also displays characteristics of germinal center reactions. Proliferating T and B lymphocytes were found within ectopic lymphoid tissue, activation-induced cytidine deaminase was expressed, and class-switched B cells were present. The presence of circular DNA intermediates, a hallmark of active class switch recombination, suggested that class switching occurs within the ectopic lymphoid tissue. Individual collections of ectopic lymphoid tissue ("lipogranulomas") from the same mouse contained different B cell repertoires, consistent with local germinal center like reactions. Class-switched anti-RNP autoantibody-producing cells were also found in the lipogranulomas. Somatic hypermutation in the lipogranulomas was T cell-dependent, as was the production of isotype-switched anti-Sm/RNP autoantibodies. Thus, ectopic lymphoid tissue induced by TMPD recapitulates many of the functional characteristics of secondary lymphoid tissue and contains autoantibody-secreting cells, which may escape from normal censoring mechanisms in this location. PMID- 19299722 TI - Modulation of T lymphocyte replicative senescence via TNF-{alpha} inhibition: role of caspase-3. AB - Expanded populations of CD8(+) T lymphocytes lacking CD28 expression are associated with a variety of deleterious clinical outcomes, including early mortality in the elderly, more rapid progression to AIDS, cardiovascular disease, and enhanced tumor cell growth. In cell culture, irreversible loss of CD28 expression correlates with increased production of TNF-alpha as CD8(+) T cells are driven to the nonproliferative end stage of replicative senescence by multiple rounds of Ag-driven cell division. Interestingly, in patients with rheumatoid arthritis, inhibition or neutralization of TNF-alpha reduces the proportion of T cells lacking CD28 in the disease joints, consistent with studies showing a direct involvement of this cytokine in CD28 gene transcription. Here, we show that modulation of TNF-alpha levels in long-term cultures of human CD8(+) T lymphocytes, by chronic exposure either to a neutralizing Ab or to an inhibitor of the TNF-alpha receptor-1, increases proliferative potential, delays loss of CD28 expression, retards cytokine profile changes, and enhances telomerase activity. We also show that constitutive caspase-3, one of the downstream effectors of TNF-alphaR1 binding, increases in parallel with the loss of CD28 in long-term cultures, but this effect is blunted in the presence of the TNF-alpha inhibitors. Consistent with the in vitro culture data, CD8(+)CD28(-) T lymphocytes tested immediately ex vivo also show significantly higher levels of caspase-3 compared with their CD28(+) counterparts. These findings help elucidate the complex nature of CD28 gene regulation, and may ultimately lead to novel therapeutic approaches for diseases associated with increased proportions of CD28(-) T lymphocytes. PMID- 19299723 TI - Recall responses by helpless memory CD8+ T cells are restricted by the up regulation of PD-1. AB - CD4 help is crucial for memory CD8(+) T cell development, yet the mechanisms of CD4 help and why (CD4) helpless memory CD8(+) T cells elicit poor recall responses are currently not well understood. In this study we investigated these questions using an in vivo acute virus infection model. We show herein that CD4 help during priming is required for memory CD8(+) T cell differentiation, and that stimulation of CD40 during priming rescues the helpless defects in the absence of CD4(+) T cells. The defective recall response by helpless memory cells did not correlate with the amount of cell death and was independent of TRAIL. However, helpless memory cells excessively up-regulated the inhibitory receptor PD-1 (programmed cell death-1), and PD-1 blockade enhanced the recall response of helpless memory cells. Furthermore, providing IL-2 signaling in vivo during the recall response reduced PD-1 expression and rescued the recall response of helpless memory cells. Our study identifies molecular pathways involved in CD4 help for memory CD8(+) T cell generation that are independent of TRAIL, and it provides therapeutic implications that helpless memory cell function can be restored at multiple stages through various immunological interventions. PMID- 19299725 TI - Uneven colonization of the lymphoid periphery by T cells that undergo early TCR{alpha} rearrangements. AB - A sparse population of thymocytes undergoes TCRalpha gene rearrangement early in development, before the double-positive stage. The potential of these cells to contribute to the peripheral T cell pool is unknown. To examine the peripheral T cell compartment expressing a repertoire biased to early TCR gene rearrangements, we developed a mouse model in which TCRalpha rearrangements are restricted to the double-negative stage of thymocyte development. These mice carry floxed RAG2 alleles and a Cre transgene driven by the CD4 promoter. As expected, conventional T cell development is compromised in such Cre(+) RAG2(fl/fl) mice, and the TCRalphabeta(+) T cells that develop are limited in their TCRalpha repertoire, preferentially using early rearranging Valpha genes. In the gut, the Thy 1(+)TCRalphabeta(+) intraepithelial lymphocyte (IEL) compartment is surprisingly intact, whereas the Thy-1(-)TCRalphabeta(+) subset is almost completely absent. Thus, T cells expressing a TCRalpha repertoire that is the product of early gene rearrangements can preferentially populate distinct IEL compartments. Despite this capacity, Cre(+) RAG2(fl/fl) T cell progenitors cannot compete with wild type T cell progenitors in mixed bone marrow chimeras, suggesting that in normal mice, there is only a small contribution to the peripheral T cell pool by cells that have undergone early TCRalpha rearrangements. In the absence of wild-type competitors, aggressive homeostatic proliferation in the IEL compartment can promote a relatively normal Thy-1(+) TCRalphabeta(+) T cell pool from the limited population derived from Cre(+) RAG2(fl/fl) progenitors. PMID- 19299724 TI - IL-7 Dependence in human B lymphopoiesis increases during progression of ontogeny from cord blood to bone marrow. AB - IL-7 is critical for B cell production in adult mice; however, its role in human B lymphopoiesis is controversial. One challenge was the inability to differentiate human cord blood (CB) or adult bone marrow (BM) hematopoietic stem cells (HSCs) without murine stroma. Here, we examine the role of IL-7 in human B cell development using a novel, human-only model based on coculturing human HSCs on primary human BM stroma. In this model, IL-7 increases human B cell production by >60-fold from both CB and adult BM HSCs. IL-7-induced increases are dose dependent and specific to CD19(+) cells. STAT5 phosphorylation and expression of the Ki-67 proliferation Ag indicate that IL-7 acts directly on CD19(+) cells to increase proliferation at the CD34(+) and CD34(-) pro-B cell stages. Without IL 7, HSCs in CB, but not BM, give rise to a small but consistent population of CD19(lo) B lineage cells that express EBF (early B cell factor) and PAX-5 and respond to subsequent IL-7 stimulation. Flt3 ligand, but not thymic stromal derived lymhopoietin (TSLP), was required for the IL-7-independent production of human B lineage cells. As compared with CB, adult BM shows a reduction of in vitro generative capacity that is progressively more profound in developmentally sequential populations, resulting in an approximately 50-fold reduction in IL-7 dependent B lineage generative capacity. These data provide evidence that IL-7 is essential for human B cell production from adult BM and that IL-7-induced expansion of the pro-B compartment is increasingly critical for human B cell production during the progression of ontogeny. PMID- 19299726 TI - Human IgG4 binds to IgG4 and conformationally altered IgG1 via Fc-Fc interactions. AB - The Fc fragment of IgG4 can interact with the Fc fragment of another IgG molecule. This interaction is a confounding factor when measuring IgG4 rheumatoid factor levels. Recently, we demonstrated that half-molecules of IgG4 can exchange to form a bispecific Ab. We expected these two phenomena to be related and investigated the physicochemical aspects of IgG4 Fc-Fc interactions. We found that IgG4 is >99% monomeric by size-exclusion chromatography; therefore, IgG4 Fc Fc interactions in the fluid phase (if any) would be short-lived. However, (125)I labeled IgG4 does bind to IgG1 and IgG4 coupled to a solid phase. By contrast, IgG1 does not bind to coupled IgG4. Furthermore, conditions that induce partial unfolding/dissociation of the CH3 domains enhance IgG4 Fc binding, suggesting that Fc binding is primarily CH3 mediated. IgG4 slowly associates with both IgG4 and IgG1 coupled to a biosensor chip. Remarkably, subsequent dissociation was much faster for IgG4 than for IgG1. Moreover, after binding of an IgG4 mAb to Sepharose-coupled Ag, we observed additional binding of IgG4 with irrelevant specificity, whereas similar binding was not observed with Ag-bound IgG1. We propose that the IgG4-IgG4 Fc interaction resembles an intermediate of the Fab arm (half-molecule) exchange reaction that is stabilized because one of the IgG4 molecules is coupled to a solid phase. By contrast, IgG4 Fc recognizes IgG1 only after a conformational change that renders CH3(IgG1) accessible to an interaction with the CH3(IgG4). Such Fc interactions may enhance Ag binding of IgG4 in vivo. PMID- 19299727 TI - A novel peptide mimotope identified as a potential immunosuppressive vaccine for organ transplantation. AB - We reported that anti-histone H1 autoantibody is one of the main immunosuppressive factors in serum that is induced after orthotopic liver transplantation in a rat tolerogenic model. We generated a novel anti-histone H1 IgM mAb produced by hybridoma 16G9 (16G9 mAb) that shows MLR-inhibitory activity. Identification of a functional epitope responsible for the immunosuppressive activity of 16G9 mAb may lead to the establishment of a novel therapeutic strategy. We used a combinatorial phage display peptide library to screen for peptides that bind to 16G9 mAb. Consequently, two peptides that bind to 16G9 mAb, SSV and LPQ, were selected from the library. The binding of 16G9 mAb to histone H1 was inhibited by SSV. SSV was recognized by rat tolerogenic post-orthotopic liver transplantation serum and the binding to SSV was inhibited by histone H1. Mice were immunized with keyhole limpet hemocyanin-conjugated SSV and LPQ. Abs induced by SSV immunization inhibited Con A-stimulated splenocyte proliferation, and the inhibition was neutralized by preincubation with SSV. Splenocytes stimulated by anti-CD3 Ab were inhibited by SSV-induced Abs using CFSE labeling. SSV immunization in rats before heterotopic heart transplantation resulted in significant prolonged allograft survival. These findings suggested that SSV is a functional histone H1-binding epitope for 16G9 mAb. SSV is capable of determining serum immunoreactivity against histone H1 as an index marker for tolerance. The inhibitory activity of SSV-induced Abs on blast cell proliferation and the prolonged graft survival that results from SSV immunization imply a potential for the development of an immunosuppressive vaccine. PMID- 19299728 TI - Vitamin d-directed rheostatic regulation of monocyte antibacterial responses. AB - The active form of vitamin D, 1,25-dihydroxyvitamin D (1,25(OH)(2)D) enhances innate immunity by inducing the cathelicidin antimicrobial peptide (hCAP). In monocytes/macrophages, this occurs primarily in response to activation of TLR, that induce expression of the vitamin D receptor and localized synthesis of 1,25(OH)(2)D from precursor 25-hydroxyvitamin D(3) (25OHD). To clarify the relationship between vitamin D and innate immunity, we assessed changes in hCAP expression in vivo and ex vivo in human subjects attending a bone clinic (n = 50). Of these, 38% were vitamin D-insufficient (<75 nM 25OHD) and received supplementation with vitamin D (50,000 IU vitamin D(2) twice weekly for 5 wk). Baseline 25OHD status or vitamin D supplementation had no effect on circulating levels of hCAP. Therefore, ex vivo changes in hCAP for each subject were assessed using peripheral blood monocytes cultured with 10% autologous serum (n = 28). Under these vitamin D "insufficient" conditions the TLR2/1 ligand 19 kDa lipopeptide or the TLR4 ligand LPS, monocytes showed increased expression of the vitamin D-activating enzyme CYP27b1 (5- and 5.5-fold, respectively, both p < 0.01) but decreased expression of hCAP mRNA (10-fold and 30-fold, both p < 0.001). Following treatment with 19 kDa, expression of hCAP: 1) correlated with 25OHD levels in serum culture supplements (R = 0.649, p < 0.001); 2) was significantly enhanced by exogenous 25OHD (5 nM); and 3) was significantly enhanced with serum from vivo vitamin D-supplemented patients. These data suggest that a key role of vitamin D in innate immunity is to maintain localized production of antibacterial hCAP following TLR activation of monocytes. PMID- 19299729 TI - TLR-induced local metabolism of vitamin D3 plays an important role in the diversification of adaptive immune responses. AB - The addition of monophosphoryl lipid A, a minimally toxic derivative of LPS, to nonmucosally administered vaccines induced both systemic and mucosal immune responses to coadministered Ags. This was dependent on an up-regulated expression of 1alpha-hydroxylase (CYP27B1, 1alphaOHase), the enzyme that converts 25 hydroxycholecalciferol, a circulating inactive metabolite of vitamin D(3), into 1,25(OH)2D(3) (calcitriol). In response to locally produced calcitriol, myeloid dendritic cells (DCs) migrated from cutaneous vaccination sites into multiple secondary lymphoid organs, including classical inductive sites of mucosal immunity, where they effectively stimulated B and T cell immune responses. The endogenous production of calcitriol by monophosphoryl lipid A-stimulated DCs appeared to be Toll-IL-1R domain-containing adapter-inducing IFN-beta-dependent, mediated through a type 1 IFN-induced expression of 1alphaOHase. Responsiveness to calcitriol was essential to promote the trafficking of mobilized DCs to nondraining lymphoid organs. Collectively, these studies help to expand our understanding of the physiologically important roles played by locally metabolized vitamin D(3) in the initiation and diversification of adaptive immune responses. The influences of locally produced calcitriol on the migration of activated DCs from sites of vaccination/infection into both draining and nondraining lymphoid organs create a condition whereby Ag-responsive B and T cells residing in multiple lymphoid organs are able to simultaneously engage in the induction of adaptive immune responses to peripherally administered Ags as if they were responding to an infection of peripheral or mucosal tissues they were designed to protect. PMID- 19299730 TI - Early NK cell-derived IFN-{gamma} is essential to host defense in neutropenic invasive aspergillosis. AB - Invasive aspergillosis is among the most common human fungal infections and occurs in patients with severe and complex defects in immune responses. NK cells have previously been found to be important in host defense against this infection, but the mechanism of this effect is not known. We hypothesized that NK cells mediate their protective effect in invasive aspergillosis by acting as the major source of IFN-gamma during early infection. We found that, in the lungs of neutropenic mice with invasive aspergillosis, NK cells were the major population of cells capable of generating IFN-gamma during early infection. Depletion of NK cells resulted in reduced lung IFN-gamma levels and increased lung fungal load that was independent of T and B cell subsets. Depletion of NK cells and absence of IFN-gamma resulted in a similar increase in susceptibility to the infection, but depletion of NK cells in IFN-gamma-deficient hosts did not result in further increase in severity of the infection. NK cell-derived IFN-gamma caused enhanced macrophage antimicrobial effects in vitro and also resulted in greater expression of IFN-inducible chemokines in the lungs. Finally, transfer of activated NK cells from wild-type, but not IFN-gamma-deficient hosts, resulted in greater pathogen clearance from the lungs of both IFN-gamma-deficient and wild-type recipients. Taken together, these data indicate that NK cells are the main source of early IFN-gamma in the lungs in neutropenic invasive aspergillosis, and this is an important mechanism in the defense against this infection. PMID- 19299731 TI - Combined effect of antiretroviral therapy and blockade of IDO in SIV-infected rhesus macaques. AB - Increased activity of IDO, which catalyzes the degradation of Trp into kynurenine (Kyn), is observed during HIV/SIV infection, and it may contribute to the persistence of HIV/SIV by suppressing antiviral T cell responses. We administered the IDO inhibitor 1-methyl-d-tryptophan (d-1mT) for 13 days to SIV-infected rhesus macaques receiving antiretroviral therapy (ART). d-1mT treatment increased the plasma levels of Trp, without reducing the levels of Kyn, suggesting only a partial effect on IDO enzymatic activity. Surprisingly, d-1mT significantly reduced the virus levels in plasma and lymph nodes of ART-treated animals with incomplete responsiveness to ART. In SIV-infected animals that were not receiving ART, d-1mT was ineffective in reducing the plasma viral load and had only a marginal effect on the plasma Kyn/Trp ratio. Increased IDO and TGF-beta mRNA expression in lymph nodes of ART-treated macaques after d-1mT treatment suggested that compensatory counterregulatory mechanisms were activated by d-1mT, which may account for the lack of effect on plasma Kyn. Finally, d-1mT did not interfere with the ART-induced T cell dynamics in lymph nodes (increased frequency of total CD4 T cells, increase of CD8 T cells expressing the antiapoptotic molecule Bcl2, and reduction of regulatory T cells). Thus, d-1mT appeared to synergize with ART in inhibiting viral replication and did not interfere with the beneficial immunologic effects of ART. Further studies are required to elucidate the immunologic or virologic mechanism by which d-1mT inhibited SIV replication in vivo. PMID- 19299732 TI - Clathrin- and dynamin-dependent endocytic pathway regulates muramyl dipeptide internalization and NOD2 activation. AB - Muramyl dipeptide (MDP), the NOD2 agonist, induces NF-kappaB and MAPK activation leading to the production of antimicrobial and proinflammatory molecules. MDP is internalized into acidified vesicles in macrophages. However, the endocytic mechanism of MDP uptake that induces NOD2 signaling is unknown. We now report the identification of an endocytosis pathway dependent on clathrin and dynamin that mediates MDP internalization and NOD2 activation. Intracellular MDP uptake was inhibited by chlorpromazine, a drug that disrupts clathrin-dependent endocytosis, but not by compounds that block pinocytosis or cellular entry via scavenger or mannose receptors. In contrast, MDP uptake and NOD2-dependent signaling were unimpaired in macrophages deficient in PepT1, a peptide transporter previously implicated in MDP internalization. Both chlorpromazine and knockdown of clathrin expression by RNA interference attenuated MDP-induced NF-kappaB and MAPK activation. Furthermore, MDP uptake and NOD2-dependent signaling were impaired by inhibition of dynamin, a GTPase required for budding of clathrin-coated vesicles from the plasma membrane. Finally, bafilomycin A, a specific inhibitor of the vacuolar proton pump, blocked MDP accumulation in acidified vesicles and cytokine responses, suggesting that vacuolar maturation is important for MDP-induced NOD2 signaling. These studies provide evidence for a clathrin- and dynamin-dependent endocytosis pathway that mediates MDP uptake and NOD2 activation. PMID- 19299733 TI - Immunologic and therapeutic synergy of IL-27 and IL-2: enhancement of T cell sensitization, tumor-specific CTL reactivity and complete regression of disseminated neuroblastoma metastases in the liver and bone marrow. AB - IL-27 exerts antitumor activity in murine orthotopic neuroblastoma, but only partial antitumor effect in disseminated disease. This study demonstrates that combined treatment with IL-2 and IL-27 induces potent antitumor activity in disseminated neuroblastoma metastasis. Complete durable tumor regression was achieved in 90% of mice bearing metastatic TBJ-IL-27 tumors treated with IL-2 compared with only 40% of mice bearing TBJ-IL-27 tumors alone and 0% of mice bearing TBJ-FLAG tumors with or without IL-2 treatment. Comparable antitumor effects were achieved by IL-27 protein produced upon hydrodynamic IL-27 plasmid DNA delivery when combined with IL-2. Although delivery of IL-27 alone, or in combination with IL-2, mediated pronounced regression of neuroblastoma metastases in the liver, combined delivery of IL-27 and IL-2 was far more effective than IL 27 alone against bone marrow metastases. Combined exposure to IL-27 produced by tumor and IL-2 synergistically enhances the generation of tumor-specific CTL reactivity. Potentiation of CTL reactivity by IL-27 occurs via mechanisms that appear to be engaged during both the initial sensitization and effector phase. Potent immunologic memory responses are generated in mice cured of their disseminated disease by combined delivery of IL-27 and IL-2, and depletion of CD8(+) ablates the antitumor efficacy of this combination. Moreover, IL-27 delivery can inhibit the expansion of CD4(+)CD25(+)Foxp3(+) regulatory and IL-17 expressing CD4(+) cells that are otherwise observed among tumor-infiltrating lymphocytes from mice treated with IL-2. These studies demonstrate that IL-27 and IL-2 synergistically induce complete tumor regression and long-term survival in mice bearing widely metastatic neuroblastoma tumors. PMID- 19299734 TI - Interactions between human NK cells and macrophages in response to Salmonella infection. AB - NK cells play a key role in host resistance to a range of pathogenic microorganisms, particularly during the initial stages of infection. NK cell interactions with cells infected with viruses and parasites have been studied extensively, but human bacterial infections have not been given the same attention. We studied crosstalk between human NK cells and macrophages infected with intracellular Salmonella. These macrophages activated NK cells, resulting in secretion of IFN-gamma and degranulation. Reciprocally, NK cell activation led to a dramatic reduction in numbers of intramacrophagic live bacteria. We identified three elements in the interaction of NK cells with infected macrophages. First, communication between NK cells and infected macrophages was contact-dependent. The second requirement was IL-2- and/or IL-15-dependent priming of NK cells to produce IFN-gamma. The third was activation of NK cells by IL-12 and IL-18, which were secreted by the Salmonella-infected macrophages. Adhesion molecules and IL 12Rbeta2 were enriched in the contact zone between NK cells and macrophages, consistent with contact- and IL-12/IL-18-dependent NK activation. Our results suggest that, in humans, bacterial clearance is consistent with a model invoking a "menage a trois" involving NK cells, IL-2/IL-15-secreting cells, and infected macrophages. PMID- 19299735 TI - The receptor for advanced glycation end products impairs host defense in pneumococcal pneumonia. AB - Streptococcus pneumoniae is the most common cause of community-acquired pneumonia. The receptor for advanced glycation end products (RAGE) is a multiligand receptor that is expressed ubiquitously in the lungs. Engagement of RAGE leads to activation of multiple intracellular signaling pathways, including NF-kappaB and subsequent transcription of several proinflammatory mediators. To determine the role of RAGE in the innate immune response to S. pneumoniae pneumonia, RAGE-deficient (RAGE(-/-)) and wild-type mice were intranasally inoculated with S. pneumoniae. S. pneumoniae pneumonia resulted in an up regulation of constitutively present RAGE expression in lung tissue, especially in the interalveolar septae. RAGE(-/-) mice showed an improved survival, which was accompanied by a lower bacterial load in the lungs at 16 h and a decreased dissemination of the bacteria to blood and spleen at 16 and 48 h after inoculation. RAGE(-/-) macrophages showed an improved killing capacity of S. pneumoniae in vitro. Lung inflammation was attenuated in RAGE(-/-) mice at 48 h after inoculation, as indicated by histopathology and cytokine/chemokine levels. Neutrophil migration to the lungs was mitigated in the RAGE(-/-) mice. In addition, in RAGE(-/-) mice, activation of coagulation was diminished. Additional studies examining the effect of RAGE deficiency on the early (6-h) inflammatory response to S. pneumoniae did not reveal an early accelerated or enhanced immune response. These data suggest that RAGE plays a detrimental role in the host response to S. pneumoniae pneumonia by facilitating the bacterial growth and dissemination and concurrently enhancing the pulmonary inflammatory and procoagulant response. PMID- 19299737 TI - Complement factor H, a marker of self protects against experimental autoimmune encephalomyelitis. AB - The CNS innate immune response is a "double-edged sword" representing a fine balance between protective antipathogen responses and detrimental neurocytotoxic effects. Hence, it is important to identify the key regulatory mechanisms involved in the control of CNS innate immunity and which could be harnessed to explore novel therapeutic avenues. In analogy to the newly described neuroimmune regulatory proteins also known as "don't eat me" signals (CD200, CD47, CD22, fractalkine, semaphorins), we herein identify the key role of complement regulator factor H (fH) in controlling neuroinflammation initiated in an acute mouse model of Ab-dependent experimental autoimmune encephalomyelitis. Mouse fH was found to be abundantly expressed by primary cultured neurons and neuronal cell lines (N1E115 and Neuro2a) at a level comparable to BV2 microglia and CLTT astrocytes. Mouse neurons expressed other complement regulators crry and low levels of CD55. In the brain, the expression of fH was localized to neuronal bodies and axons, endothelial cells, microglia but not oligodendrocytes and myelin sheaths and was dramatically reduced in inflammatory experimental autoimmune encephalomyelitis settings. When exogenous human fH was administered to disease Ab-dependent experimental autoimmune encephalomyelitis animals, there was a significant decrease in clinical score, inflammation, and demyelination, as compared with PBS-injected animals. We found that the accumulation of human fH in the brain parenchyma protected neurons from complement opsonization, axonal injury, and leukocyte infiltration. Our data argue for a key regulatory activity of fH in neuroprotection and provide novel therapeutic avenues for CNS chronic inflammatory diseases. PMID- 19299736 TI - Airway and lung pathology due to mucosal surface dehydration in {beta}-epithelial Na+ channel-overexpressing mice: role of TNF-{alpha} and IL-4R{alpha} signaling, influence of neonatal development, and limited efficacy of glucocorticoid treatment. AB - Overexpression of the epithelial Na(+) channel beta subunit (Scnn1b gene, betaENaC protein) in transgenic (Tg) mouse airways dehydrates mucosal surfaces, producing mucus obstruction, inflammation, and neonatal mortality. Airway inflammation includes macrophage activation, neutrophil and eosinophil recruitment, and elevated KC, TNF-alpha, and chitinase levels. These changes recapitulate aspects of complex human obstructive airway diseases, but their molecular mechanisms are poorly understood. We used genetic and pharmacologic approaches to identify pathways relevant to the development of Scnn1b-Tg mouse lung pathology. Genetic deletion of TNF-alpha or its receptor, TNFR1, had no measurable effect on the phenotype. Deletion of IL-4Ralpha abolished transient mucous secretory cell (MuSC) abundance and eosinophilia normally observed in neonatal wild-type mice. Similarly, IL-4Ralpha deficiency decreased MuSC and eosinophils in neonatal Scnn1b-Tg mice, which correlated with improved neonatal survival. However, chronic lung pathology in adult Scnn1b-Tg mice was not affected by IL-4Ralpha status. Prednisolone treatment ablated eosinophilia and MuSC in adult Scnn1b-Tg mice, but did not decrease mucus plugging or neutrophilia. These studies demonstrate that: 1) normal neonatal mouse airway development entails an IL-4Ralpha-dependent, transient abundance of MuSC and eosinophils; 2) absence of IL-4Ralpha improved neonatal survival of Scnn1b-Tg mice, likely reflecting decreased formation of asphyxiating mucus plugs; and 3) in Scnn1b-Tg mice, neutrophilia, mucus obstruction, and airspace enlargement are IL-4Ralpha- and TNF-alpha-independent, and only MuSC and eosinophilia are sensitive to glucocorticoids. Thus, manipulation of multiple pathways will likely be required to treat the complex pathogenesis caused by airway surface dehydration. PMID- 19299738 TI - Activation of microglia by amyloid {beta} requires P2X7 receptor expression. AB - Extracellular ATP is a mediator of intercellular communication and a danger signal. Release of this and other nucleotides modulates microglia responses via P2Y and P2X receptors, among which the P2X(7) subtype stands out for its proinflammatory activity and for up-regulation in a transgenic model of Alzheimer disease and in brains from Alzheimer disease patients. Here we show that amyloid beta (Abeta) triggered increases in intracellular Ca(2+) ([Ca(2+)](i)), ATP release, IL-1beta secretion, and plasma membrane permeabilization in microglia from wild-type but not from P2X(7)-deleted mice. Likewise, intra-hippocampal injection of Abeta caused a large accumulation of IL-1beta in wild-type but not in P2X(7)(-/-) mice. These observations suggest that Abeta activates a purinergic autocrine/paracrine stimulatory loop of which the P2X(7) receptor is an obligate component. Identification of the P2X(7) receptor as a non-dispensable factor of Abeta-mediated microglia stimulation may open new avenues for the treatment of Alzheimer disease. PMID- 19299739 TI - Bacterial DNA activates endothelial cells and promotes neutrophil adherence through TLR9 signaling. AB - TLR9 detects bacterial DNA (CpG DNA) and elicits both innate and adoptive immunity. Recent evidence indicates that TLR9 is expressed in more diverse cell types than initially thought. In this study, we report that HUVECs constitutively express TLR9 and selectively recognize unmethylated CpG motifs in bacterial DNA and synthetic immune stimulatory CpG oligodeoxynucleotides. HUVECs respond to CpG DNA with rapid phosphorylation of IkappaB-alpha and NF-kappaB-mediated gene transcription and surface expression of the adhesion molecules ICAM-1 and E selectin independent of MAPK signaling. The telomere-derived TLR9 inhibitory oligonucleotide 5'-TTT AGG GTT AGG GTT AGG G-3', agents that block endosomal acidification such as chloroquine and bafilomycin A, and NF-kappaB inhibitors abrogated CpG DNA-induced signaling. HUVEC activation by CpG DNA led to markedly enhanced neutrophil adhesion under nonstatic conditions that was further enhanced when neutrophils were stimulated with CpG DNA. The adhesive interactions were blocked by Abs against CD18 and, to a lesser degree, by anti-E-selectin and anti L-selectin Abs. Our findings demonstrate that bacterial DNA promotes beta(2) integrin and E-selectin-mediated HUVEC-neutrophil adherence, and indicate the ability of CpG DNA to initiate and/or maintain the inflammatory response. PMID- 19299740 TI - Isoforms of vitamin E have opposing immunoregulatory functions during inflammation by regulating leukocyte recruitment. AB - Reports indicate contradictory outcomes for anti-inflammatory functions of the alpha-tocopherol isoform of vitamin E in clinical studies of asthma and atherosclerosis. These seemingly disparate clinical results are consistent with novel unrecognized properties of isoforms of vitamin E reported in this study. We demonstrate that the isoform d-gamma-tocopherol elevates inflammation in experimental asthma. Moreover, d-gamma-tocopherol, at as little as 10% the concentration of d-alpha-tocopherol, ablates the anti-inflammatory benefit of the d-alpha-tocopherol isoform. A mechanism for these opposing immunoregulatory functions of purified tocopherols at physiological concentrations is not through modulation of expression of several cytokines, chemokines, or adhesion molecules, but is, at least in part, by regulation of endothelial cell signals during leukocyte recruitment. These opposing regulatory functions of vitamin E isoforms have impact on interpretations of vitamin E studies. In summary, our studies with purified tocopherol isoforms alter our understanding of vitamin E regulation of vascular function and asthma. PMID- 19299741 TI - Mechanism of the salutary effects of estrogen on kupffer cell phagocytic capacity following trauma-hemorrhage: pivotal role of Akt activation. AB - Kupffer cells are macrophages in the liver whose major role is to clear circulating pathogens. Decreased phagocytic capacity of Kupffer cells may result in severe systemic infection. We tested the hypothesis that the depressed Kupffer cell phagocytic capacity following trauma-hemorrhage is enhanced by estrogen administration and this occurs due to maintenance of Fc receptor expression and cellular ATP content via the activation of Akt. Male C3H/HeN mice were subjected to sham operation or trauma-hemorrhage and sacrificed 2 h thereafter. Estrogen, with or without an estrogen receptor antagonist (ICI 182,780), a PI3K inhibitor (Wortmannin), or vehicle, was injected during resuscitation. Kupffer cell phagocytic capacity was tested in vivo. The expression of Fc receptors, of Akt phosphorylation, of p38 MAPK phosphorylation, of DNA binding activity of NF kappaB and ATP content of Kupffer cells were also determined. Trauma-hemorrhage suppressed Kupffer cell phagocytosis by decreasing Fc receptor expression and Akt activation; however, it induced p38 MAPK activation and increased NF-kappaB activity. Cellular ATP levels were also decreased following trauma-hemorrhage. Administration of estrogen following trauma-hemorrhage increased phospho-Akt levels and normalized all the parameters described as well as plasma levels of TNF-alpha, IL-6, and IL-10. Coadministration of ICI 182,780 or Wortmannin abolished the beneficial effects of estrogen in improving the phagocytic capacity of Kupffer cells following trauma-hemorrhage. Thus, activation of Akt plays a crucial role in mediating the salutary effect of estrogen in restoring trauma hemorrhage-induced suppression of Kupffer cell phagocytosis. PMID- 19299742 TI - M2 macrophages phagocytose rituximab-opsonized leukemic targets more efficiently than m1 cells in vitro. AB - Because macrophages have been implicated as major players in the mechanism of action of rituximab, we have investigated the factors that modulate their tumor cell killing potential. Human macrophages, differentiated in vitro from peripheral blood monocytes, were used in binding and phagocytosis assays using B chronic lymphocytic leukemia or lymphoma target cells opsonized with rituximab. Phagocytosis was maximal at 0.1 microg/ml rituximab and was not significantly affected by CD20 expression levels or by CD16A polymorphism at position 158 (Val/Phe). The role of FcgammaRs was demonstrated by complete inhibition of phagocytosis by excess human Igs. Because macrophages can be differentiated to M1 or M2-type cells with either GM-CSF or M-CSF, respectively, and can be classically activated by proinflammatory stimuli (IFN-gamma/LPS) or undergo alternative activation with cytokines such as IL-4 or IL-10, we have analyzed the effect of these different polarization programs on the phagocytosis mediated by rituximab. Macrophages differentiated in presence of M-CSF showed a 2- to 3-fold greater phagocytic capacity compared with GM-CSF-induced cells. Furthermore, addition of IL-10 significantly increased, whereas IL-4 decreased phagocytosis by both M-CSF- and GM-CSF-differentiated macrophages. LPS/IFN-gamma had little effect. Expression of CD16, CD32, and CD64 in different macrophage populations correlated with phagocytic activity. Interestingly, several B lymphoma cell lines were observed to secrete 400-1300 pg/ml IL-10 in vitro, and coculture of human macrophages with lymphoma conditioned medium increased significantly their phagocytic capacity. Our data suggest that cytokines secreted by lymphoma cells can favor alternate activation of macrophages with a high phagocytic capacity toward rituximab-opsonized targets. PMID- 19299743 TI - The presence of a matrix-derived neutrophil chemoattractant in bronchiolitis obliterans syndrome after lung transplantation. AB - Lung transplantation is a therapeutic modality frequently used in end-stage lung disease. Unfortunately, lung transplant recipients have poor clinical outcomes, often due to the development of bronchiolitis obliterans syndrome (BOS). This process is often characterized by the pathologic findings of obliterative bronchiolitis: neutrophil influx and extracellular matrix remodeling leading to luminal obstruction and airway inflammation. The molecular mechanisms underlying BOS are poorly understood and disease-specific biomarkers are lacking. We report that in addition to increased levels of IL-8, the level of the neutrophil chemoattractant proline-glycine-proline (PGP) is elevated in BOS patient bronchoalveolar lavage (BAL) fluid. The enzymes responsible for generating PGP, matrix metalloproteases 8 and -9 and prolyl endopeptidase, are also elevated in these samples. Together, IL-8 and PGP account for most of the neutrophil chemoattractant capacity seen in BOS BAL fluid. Using specific neutralizing Abs to both IL-8 and PGP, we demonstrate that PGP is a prominent neutrophil chemoattractant found in BAL fluid from individuals at the time of diagnosis of BOS. These findings highlight the influence of a matrix-derived neutrophil chemoattractant in posttransplantation BOS and provide opportunities for the development of unique diagnostics and therapeutics to potentially improve disease outcomes. PMID- 19299744 TI - Advanced intercross line mapping suggests that ncf1 (ean6) regulates severity in an animal model of guillain-barre syndrome. AB - We here present the first genetic fine mapping of experimental autoimmune neuritis (EAN), the animal model of Guillain-Barre syndrome, in a rat advanced intercross line. We identified and refined a total of five quantitative trait loci on rat chromosomes 4, 10, and 12 (RNO4, RNO10, RNO12), showing linkage to splenic IFN-gamma secretion and disease severity. All quantitative trait loci were shared with other models of complex inflammatory diseases. The quantitative trait locus showing strongest linkage to clinical disease was Ean6 and spans 4.3 Mb on RNO12, harboring the neutrophil cytosolic factor 1 (Ncf1) among other genes. Polymorphisms in Ncf1, a member of the NADPH oxidase complex, have been associated with disease regulation in experimental arthritis and encephalomyelitis. We therefore tested the Ncf1 pathway by treating rats with a NADPH oxidase complex activator and ameliorated EAN compared the oil-treated control group. By proving the therapeutic effect of stimulating the NADPH oxidase complex, our data strongly suggest the first identification of a gene regulating peripheral nervous system inflammation. Taken together with previous reports, our findings suggest a general role of Ncf1 and oxidative burst in pathogenesis of experimental autoimmune animal models. PMID- 19299745 TI - Subtle affinity-enhancing mutations in a myelin oligodendrocyte glycoprotein specific TCR alter specificity and generate new self-reactivity. AB - We describe a simple iterative approach to augment TCR affinity, which we studied using a myelin oligodendrocyte glycoprotein-specific TCR. We hypothesized that single amino acid modifications in TCR CDR3 could enhance TCR sensitivity through focal interactions with antigenic peptide while minimizing the risk of cross reactivity observed previously in TCR more broadly mutagenized using in vitro evolution techniques. We show that this iterative method can indeed generate TCR with Ag sensitivity 100-fold greater than the parental receptor and can endow TCR with coreceptor independence. However, we also find that single amino acid mutations in the CDR3 can alter TCR fine specificity, affecting recognition requirements for Ag residues over most of the length of the MHC binding groove. Furthermore, minimal changes in surface-exposed CDR3 amino acids, even the addition of a single hydroxyl group or conversion of a methyl or sulfhydryl moiety to a hydroxyl, can confer modified Ag-specific TCR with new self reactivity. In vivo modeling of modified TCR through retroviral TCR gene transfer into Rag(-/-) mice confirmed the biological significance of these altered reactivities, although it also demonstrated the feasibility of producing Ag specific, positively selecting, coreceptor-independent receptors with markedly increased Ag sensitivity. These results affirm the possibility of readily generating affinity-enhanced TCR for therapeutic purposes but demonstrate that minimal changes in TCR CDR3 structure can promote self reactivity and thereby emphasize the importance of caution in validating receptors with even subtle alterations before clinical application. PMID- 19299746 TI - CXCR4/CXCL12 hyperexpression plays a pivotal role in the pathogenesis of lupus. AB - Among various surface molecules screened, CXCR4 was significantly up-regulated on monocytes, neutrophils, B cell subsets, and plasma cells in multiple murine models of lupus with active nephritis, including B6.Sle1Yaa, BXSB, and MRL.lpr. TLR-mediated signaling and inflammatory cytokines accounted in part for this increase. Increased CXCR4 expression was associated with functional consequences, including increased migration and enhanced B cell survival. Simultaneously, the ligand for CXCR4, CXCL12, was significantly up-regulated in the nephritic kidneys. Treatment with a peptide antagonist of CXCR4 prolonged survival and reduced serum autoantibodies, splenomegaly, intrarenal leukocyte trafficking, and end organ disease in a murine model of lupus. These findings underscore the pathogenic role of CXCR4/CXCL12 in lymphoproliferative lupus and lupus nephritis and highlight this axis as a promising therapeutic target in this disease. PMID- 19299748 TI - TLR3 ligand induces NF-{kappa}B activation and various fates of multiple myeloma cells depending on IFN-{alpha} production. AB - Multiple myeloma (MM) cells express TLR. It has been shown that TLR ligands induce the proliferation, survival, and immune surveillance escape of MM cells through MyD88-TLR pathways. Deciphering TLR function in MM cells will help in understanding the mechanisms of tumor cell growth. In this study, we examined the response of MM cells to the MyD88-independent/TIR-domain-containing adapter inducing IFN-beta-dependent TLR3. Deregulation of NF-kappaB pathway is a feature of MM cells, and we wondered whether TLR3 activation could mobilize the NF-kappaB pathway. We show that five of seven human myeloma cell line (HMCL) cells expressed TLR3. In the presence of the synthetic TLR3 ligand (poly(I:C)), activation of NF-kappaB pathway was observed in three of five selected TLR3(+) HMCL, NCI-H929, RPMI 8226, and KMM1. In agreement with NF-kappaB activation, only these three HMCL responded to poly(I:C), although by either an increase (KMM1) or a decrease (NCI-H929, RPMI 8226) of proliferation. We show that KMM1 increase of proliferation was prevented by NF-kappaB inhibitor. In contrast, inhibition of proliferation in both NCI-H929 and RPMI 8226 was due to IFN-alpha-induced apoptosis. We next demonstrated that p38 MAPK pathway controlled both IFN-alpha secretion and IFN-alpha-mediated cell death. Moreover, cell death also involved activation of ERK1/2 pathway. In conclusion, our results show that TLR3 ligand induces NF-kappaB pathway activation in MM and support a switching function of type I IFN in the functional outcome of TLR3 triggering in tumor cells. PMID- 19299747 TI - Circulating monocytes in HIV-1-infected viremic subjects exhibit an antiapoptosis gene signature and virus- and host-mediated apoptosis resistance. AB - Mechanisms that may allow circulating monocytes to persist as CD4 T cells diminish in HIV-1 infection have not been investigated. We have characterized steady-state gene expression signatures in circulating monocytes from HIV infected subjects and have identified a stable antiapoptosis gene signature comprised of 38 genes associated with p53, CD40L, TNF, and MAPK signaling networks. The significance of this gene signature is indicated by our demonstration of cadmium chloride- or Fas ligand-induced apoptosis resistance in circulating monocytes in contrast to increasing apoptosis in CD4 T cells from the same infected subjects. As potential mechanisms in vivo, we show that monocyte CCR5 binding by HIV-1 virus or agonist chemokines serves as independent viral and host modulators resulting in increased monocyte apoptosis resistance in vitro. We also show evidence for concordance between circulating monocyte apoptosis-related gene expression in HIV-1 infection in vivo and available datasets following viral infection or envelope exposure in monocyte-derived macrophages in vitro. The identification of in vivo gene expression associated with monocyte resistance to apoptosis is of relevance to AIDS pathogenesis since it would contribute to: 1) maintaining viability of infection targets and long-term reservoirs of HIV-1 infection in the monocyte/macrophage populations, and 2) protecting a cell subset critical to host survival despite sustained high viral replication. PMID- 19299750 TI - Exploring transitions. PMID- 19299749 TI - Transcriptional modulation of the immune response by peroxisome proliferator activated receptor-{alpha} agonists in autoimmune disease. AB - Peroxisome proliferator-activated receptor-alpha (PPARalpha) agonists have been shown to have a therapeutic benefit in experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis (MS). In this study, we investigated the mechanism by which the PPARalpha agonist gemfibrozil induces immune deviation and protects mice from EAE. We demonstrated that treatment with gemfibrozil increases expression of the Th2 transcription factor GATA-3 and decreases expression of the Th1 transcription factor T-bet in vitro and directly ex vivo. These changes correlated with an increase in nuclear PPARalpha expression. Moreover, the protective effects of PPARalpha agonists in EAE were shown to be partially dependent on IL-4 and to occur in a receptor-dependent manner. PPARalpha was demonstrated, for the first time, to regulate the IL-4 and IL-5 genes and to bind the IL-4 promoter in the presence of steroid receptor coactivator-1, indicating that PPARalpha can directly transactivate the IL-4 gene. Finally, therapeutic administration of PPARalpha agonists ameliorated clinically established EAE, suggesting that PPARalpha agonists may provide a treatment option for immune-mediated inflammatory diseases. PMID- 19299751 TI - Pathologizing or validating: intake workers' discursive constructions of mothers. AB - We employ a feminist discursive lens to re-examine an earlier thematic-based analysis of intake workers' reports of interviews with mothers at a children's hospital. In the original study the first author focused on coping styles of parents of children with exceptionalities, whereas in the current study we demonstrate how intake workers' discursive constructions of mothers as pathological functions to dismiss mothers' views and concerns, and constructions of sets of parents as open to expert advice functions to validate them as "effective allies." In this article, we seek to provide an example of how our analytical lens serves to produce various findings that might serve the voices of the more powerful, or of the marginalized. As an outcome of the re-analysis of the data, we recommend an approach to parent- professional interaction that involves enacting language that fosters mutual respect between professionals and parents, and in particular between intake workers and mothers. PMID- 19299752 TI - Illness meanings of AIDS among women with HIV: merging immunology and life experience. AB - Little attention has been paid to illness meanings held by those with HIV in the presence of highly active antiretroviral therapy (HAART). This article illustrates how elucidating illness meanings might aid our understanding of HAART adherence and other important health behaviors. Using multiple qualitative methods, including in-depth interviews, free lists, and drawings, I explore meanings surrounding the concept of AIDS among women with HIV in New Orleans, Louisiana. Illness meanings of AIDS reflect the women's negotiation of physical, social, and emotional threats posed by HIV. HIV-positive women displace death and stigma away from an HIV diagnosis to an AIDS diagnosis, creating a sense of safety and normalcy for themselves. Women with AIDS diagnoses, however, struggle to construct illness meanings of AIDS that resist its association with ostracism and death. Drawings produced by the women provide rich insights and illustrate the value of drawings as a visual method in exploring illness meanings. PMID- 19299753 TI - Muscle gains and emotional strains: conflicting experiences of change among overweight women participating in an exercise intervention program. AB - In this study we explored the experiences of women who were classified as overweight while they participated in a physical activity intervention. In line with interpretative phenomenological analysis, eight women were interviewed prior to and following a 12-week dragon boat physical activity intervention. Data were transcribed verbatim and subjected to individual-level content analysis to explore change that informed group-level idiographic analysis. From the idiographic analysis three distinct profiles were created: (a) women who consistently struggled with negative self-perceptions; (b) women who consistently experienced positive self-perceptions; and (c) women who began with negative self perceptions and developed more positive self-images. These profiles appeared to be associated with age, since the youngest women placed significant emphasis on the body and physical appearance, whereas the oldest participants reported the most significant shift from importance of body to a greater emphasis on health and well-being. The findings, which are reported in the context of self determination theory, suggest that women in each of these profiles had unique physical and social self-perceptions and distinct social interactions within the dragon boat intervention. PMID- 19299754 TI - Reproductive decisions for women with HIV: motherhood's role in envisioning a future. AB - Cultural influences might exert more influence on HIV-positive women's reproductive choices than HIV-related conditions. In this article we report on grounded theory research on how women with HIV made reproductive decisions during a time of transition from HIV as potentially fatal to mothers and newborns to its current status as an often controllable chronic illness. Eighty HIV-positive women of childbearing age in three United States cities were interviewed, and the interviews were analyzed using grounded theory techniques. The core concept of the findings was that women's decisions were based on their judgment of the relative weight of positive aspects of motherhood versus the often negative pressures of social and public opinion. These findings have relevance for future research as well as for program development and policy. PMID- 19299755 TI - The influence of family and culture on physical activity among female adolescents from the Indian diaspora. AB - In this study we explored the role of personal, familial, and cultural attitudes and social norms for physical activity (PA) on actual PA behavior among female adolescents of the Indian diaspora. Six girls, 15 to 19 years of age, from a spiritual center participated in interviews and a focus group. Data were analyzed using content analysis. Participants were high in familism, and felt that PA was important for physical and mental health, and to strengthen relationships with family. Fathers and brothers were considered most influential on PA patterns. Differentiated gender roles in PA emerged: boys were deemed more aggressive and competitive, and girls were perceived to promote fun-based learning environments. The importance of religion and spirituality as influences on PA emerged among participants with strong affinities for Indian cultures. Results show that cultural heritage impacts PA norms, attitudes, and patterns, and must be considered when evaluating adolescent PA participation in multicultural societies. PMID- 19299756 TI - Personal values and meaning in the use of methamphetamine among HIV-positive men who have sex with men. AB - Our aim with this qualitative study was to understand the role of personal values, meaning, and impact of drug use among HIV-positive men who have sex with men (MSM) who struggle with methamphetamine use. Participants were 22 MSM recruited from an ethnically diverse county in the San Francisco Bay area of California. Grounded theory was used to analyze the data collected in individual interviews. Emergent constructs of context, meaning, and perceived impact were identified and are described in a theoretical narrative format. The importance of broadening our understanding of HIV and methamphetamine addiction and their interaction is highlighted. This study contributes to the understanding of the complexity of methamphetamine use within the specific population of MSM living with HIV/ AIDS, and suggests possible directions for addressing important maintaining factors like adaptive use and enhancing factors that could contribute to an individual's ability to make better choices based on meaning and personal values. PMID- 19299757 TI - Influencing health debates through letters to the editor: the case of male circumcision. AB - In this article I use the case of male circumcision (MC) to examine how grassroots activists, medical professionals, other stakeholders, and ordinary people employ letters to the editor (LTEs) to influence public health debates. I also show how journalistic practices affect the use of LTEs. Seventy LTEs about MC from U.S. newspapers between 1985 and 2006 are analyzed using qualitative methods. Pro-MC, anti-MC, and neutral LTE writers supported their stances on similar grounds, described adversaries as biased, and stressed medical and scientific authority. Yet only MC advocates and neutralists trivialized MC and declined to justify their stances, suggesting distinctive dynamics for LTEs about widely accepted practices. The prevalence of debated practices and activists' efforts to piggyback on related issues also affect LTE content. Editors chose LTEs to address readers' critiques, enact news values like balance and controversy, and showcase writers with strong claims to legitimacy, thereby mediating public health debates. PMID- 19299758 TI - Barriers to service use for postpartum depression symptoms among low-income ethnic minority mothers in the United States. AB - The risks of untreated postpartum depression (PPD) in the United States are higher among low-income ethnic minority mothers. However, research has not adequately investigated barriers to formal help seeking for PPD symptoms among this vulnerable population. We used convenience and purposive sampling strategies to recruit mothers experiencing past-year (the year prior to interview) PPD symptoms (n=14), community key informants (n=11), and service providers (n=12) to participate in focus groups and individual interviews. A grounded theory analysis of these nested perspectives revealed individual, community, and provider-level barriers operating at various stages of the help-seeking process: thinking about symptoms, seeking advice, and rejecting formal care. Although mothers overwhelmingly recommended "talking it out" for other mothers with PPD, an array of attitudinal and instrumental barriers led mothers to choose self-help practices in lieu of formal mental health care. PMID- 19299759 TI - Evaluating internet interviews with gay men. AB - In this article we describe the use of the Internet to conduct online synchronous interviews on gay men's experiences of Internet sex-seeking and barebacking, using interpretative phenomenological analysis (IPA). We then present the results of a separate IPA of the issues we encountered during the research. The analysis is focused around four superordinate themes: positive experience, technological issues, boundaries, and requests/expectations. We argue that online qualitative research is valuable in its own right, and that the advantages considerably outweigh the difficulties. This is particularly true in accessing individuals who might not participate in other types of research settings to discuss experiences that would otherwise be too difficult to talk about. Several issues that are problematic in more conventional research still apply, albeit in a different guise, but these must be situated in the context of who or what is being researched, and why, and can be largely overcome with more creative methodologies and considered preparation. PMID- 19299760 TI - Near-infrared spectroscopy: shedding light on the injured brain. PMID- 19299761 TI - All results count. PMID- 19299762 TI - Limits to lipid in the literature and lab: what we know, what we don't know. PMID- 19299763 TI - A preliminary report on the prognostic significance of preoperative brain natriuretic peptide and postoperative cardiac troponin in patients undergoing major vascular surgery. AB - BACKGROUND: Associations between preoperative elevation of brain natriuretic peptide (BNP) or postoperative elevation of cardiac troponins (cTn) with major adverse cardiac events (MACE) after major surgery have been shown previously. In this study, we evaluated the added value of preoperative BNP with postoperative cTn levels for the prediction of MACE in patients undergoing major vascular surgery. METHODS: This is a prospectively prespecified, secondary analysis of data from a cohort of 133 clinically stable patients undergoing major vascular surgery enrolled in a clinical trial evaluating the effectiveness of the sympathetic nervous system-inhibiting drug moxonidine on reducing MACE. Concentrations of BNP and cTn were determined before surgery, and concentrations of cTn were measured immediately after surgery and on postoperative days 1, 2, 3, and 7. The primary end point was the occurrence of MACE (defined as any hospitalization for myocardial revascularization, acute coronary syndrome, acute congestive heart failure, or death by any cause) within 1 yr after surgery. Patients were evaluated for MACE by hospital chart review during hospitalization and by telephone interviews 12 mo after surgery. RESULTS: Within 1 yr after surgery, 19 patients (14%) had a MACE, including 14 patients (11%) who died. After adjustment for age, gender, and the revised cardiac risk index, preoperative BNP elevation > or =50 pg/mL was associated with MACE (adjusted hazard ratio [HR]: 6.5, 95% confidence interval [CI]: 1.4-29.5) regardless of the subsequent cTn I concentrations. The combination of preoperative BNP elevation > or =50 pg/mL and postoperative cTn I elevation > or =2 ng/mL was associated with MACE (adjusted HR: 25.2, 95% CI: 5.0-128.4) and all-cause mortality (adjusted HR: 18.7, 95% CI: 3.1-112.5). The negative predictive value of a normal preoperative BNP value for subsequent adverse events was 0.965 (95% CI: 0.879-0.996). CONCLUSION: These data suggest that measurement of preoperative BNP concentrations in addition to postoperative cTn concentrations provides additive prognostic information for MACE and mortality after major vascular surgery. PMID- 19299764 TI - Transient metabolic alkalosis during early reperfusion abolishes helium preconditioning against myocardial infarction: restoration of cardioprotection by cyclosporin A in rabbits. AB - BACKGROUND: Intracellular acidosis during early reperfusion after coronary artery occlusion was recently linked to cardioprotection resulting from myocardial ischemic postconditioning. We tested the hypotheses that transient alkalosis during early reperfusion abolishes helium preconditioning and that the mitochondrial permeability transition pore inhibitor cyclosporin A (CsA) restores the cardioprotective effects of helium during alkalosis in vivo. METHODS: Rabbits (n = 36) instrumented for hemodynamics measurement were subjected to a 30-min left anterior descending coronary artery occlusion and 3-h reperfusion. The rabbits received 0.9% saline (control) or three cycles of 70% helium-30% oxygen administered for 5 min interspersed with 5 min of an air-oxygen mixture before left anterior descending coronary artery occlusion in the absence or presence of transient alkalosis (pH = 7.5) produced by administration of IV sodium bicarbonate (10 mEq) 2 min before reperfusion. Other rabbits preconditioned with helium received CsA (5 mg/kg) in the presence of alkalosis or CsA alone. RESULTS: Helium reduced myocardial infarct size (25% +/- 4% of left ventricular area at risk; P < 0.05) compared with control (44% +/- 6%). Alkalosis during early reperfusion did not alter infarct size alone (46% +/- 2%), but this intervention abolished helium-induced cardioprotection (45% +/- 3%). CsA restored reductions in infarct size produced by helium preconditioning in the presence of alkalosis (28% +/- 6%; P < 0.05 versus control) but did not affect myocardial necrosis alone (43% +/- 6%). CONCLUSIONS: The results demonstrate that transient alkalosis during early reperfusion abolishes helium preconditioning in rabbits. CsA restored helium-induced cardioprotection during alkalosis, suggesting that helium preconditioning inhibits mitochondrial permeability transition pore formation by maintaining intracellular acidosis during early reperfusion. PMID- 19299765 TI - Platelet transfusion during liver transplantation is associated with increased postoperative mortality due to acute lung injury. AB - BACKGROUND: Platelet transfusions have been identified as an independent risk factor for survival after orthotopic liver transplantation (OLT). In this study, we analyzed the specific causes of mortality and graft loss in relation to platelet transfusions during OLT. METHODS: In a series of 449 consecutive adult patients undergoing a first OLT, the causes of patient death and graft failure were studied in patients who did or did not receive perioperative platelet transfusions. RESULTS: Patient and graft survival were significantly reduced in patients who received platelet transfusions, compared with those who did not (74% vs 92%, and 69% vs 85%, respectively at 1 yr; P < 0.001). Lower survival rates in patients who received platelets were attributed to a significantly higher rate of early mortality because of acute lung injury (4.4% vs 0.4%; P = 0.004). There were no significant differences in other causes of mortality between the two groups. The main cause of graft loss in patients receiving platelets was patient death with a functioning graft. CONCLUSIONS: These findings suggest that platelet transfusions are an important risk factor for mortality after OLT. The current study extends previous observations by identifying acute lung injury as the main determinant of increased mortality. The higher rate of graft loss in patients receiving platelets is related to the higher overall mortality rate and does not result from specific adverse effects of transfused platelets on the grafted liver. PMID- 19299766 TI - The use of air in the inspired gas mixture during two-lung ventilation delays lung collapse during one-lung ventilation. AB - BACKGROUND: Collapse of the ipsilateral lung facilitates surgical exposure during thoracic procedures. The use of different gas mixtures during two-lung ventilation (2LV) may improve or impede surgical conditions during subsequent one lung ventilation (OLV) by increasing or delaying lung collapse. We investigated the effects of three different gas mixtures during 2LV on lung collapse and oxygenation during subsequent OLV: Air/Oxygen (fraction of inspired oxygen [FIO(2)] = 0.4), Nitrous Oxide/Oxygen ("N(2)O," FIO(2) = 0.4) and Oxygen ("O(2)," FIO(2) = 1.0). METHODS: Subjects were randomized into three groups: Air/Oxygen (n = 33), N(2)O (n = 34) or O(2) (n = 33) and received the designated gas mixture during induction and until the start of OLV. Subjects' lungs in all groups were then ventilated with FIO(2) = 1.0 during OLV. The surgeons, who were blinded to the randomization, evaluated the lung deflation using a verbal rating scale at 10 and 20 min after the start of OLV. Serial arterial blood gases were performed before anesthesia induction, during 2LV, and every 5 min, for 30 min, after initiation of OLV. RESULTS: The use of air in the inspired gas mixture during 2LV led to delayed lung deflation during OLV, whereas N(2)O improved lung collapse. Arterial oxygenation was significantly improved in the O(2) group only for the first 10 min of OLV, after which there were no differences in mean Pao(2) values among groups. CONCLUSIONS: De-nitrogenation of the lung during 2LV is a useful strategy to improve surgical conditions during OLV. The use of FIO(2) 1.0 or N(2)O/O(2) (FIO(2) 0.4) during 2LV did not have an adverse effect on subsequent oxygenation during OLV. PMID- 19299767 TI - Choosing a lung isolation device for thoracic surgery: a randomized trial of three bronchial blockers versus double-lumen tubes. AB - BACKGROUND: There is no consensus on the best technique for lung isolation for thoracic surgery. In this study, we compared the clinical performance of three bronchial blockers (BBs) available in North America with left-sided double-lumen tubes (DLTs) for lung isolation in patients undergoing left-sided thoracic surgery. METHODS: One hundred four patients undergoing left-sided thoracotomy or video-assisted thoracoscopic surgery were randomly assigned to one of the four lung isolation groups (n = 26/group). Lung isolation was with an Arndt wire guided BB (Cook Critical Care, Bloomington, IN), a Cohen Flexi-tip BB (Cook Critical Care) or a Fuji Uni-blocker (Fuji Systems, Tokyo) or with a left-sided DLT (Mallinckrodt Medical, Cornamadde, Athlone, Westmeath, Ireland). Anesthetic management and lung isolation were performed according to a standardized protocol. Each group was randomly subdivided into two subgroups (n = 13/subgroup): immediate suction (at the time of insertion of the lung isolation device) (Subgroup I) or delayed suction (20 min after insertion of the lung separation device) (Subgroup D) according to when suction was applied to the BB suction channel or the bronchial lumen of the DLT. Using a verbal analog scale, lung collapse was assessed by the surgeons, who were blinded to the lung isolation technique. RESULTS: There was no difference among the lung isolation devices in lung collapse scores at 0 (P = 0.66), 10 (P = 0.78), or 20 min (P = 0.51) after pleural opening. The time to initial lung isolation was less for DLTs (93 +/- 62 s) than BBs (203 +/- 132) (P = 0.0001). There were no differences among the BBs in the time to lung isolation (P = 0.78). There were significantly more repositions after initial placement of the lung isolation device with BBs (35 incidents) than with DLTs (two incidents) (P = 0.009). The Arndt BB required repositioning more frequently (16 incidents) than the Cohen BB (8) or the Fuji BB (11) (P = 0.032). CONCLUSIONS: The three BBs provided equivalent surgical exposure to left-sided DLTs during left-sided open or video-assisted thoracoscopic surgery thoracic procedures. BBs required longer to position and required intraoperative repositioning more often. The Arndt BB needed to be repositioned more often than the other BBs. PMID- 19299768 TI - Systolic anterior motion as a result of relative "undersizing" of a mitral valve annulus in a patient with Barlow's disease. PMID- 19299769 TI - A case of left ventricular noncompaction. PMID- 19299770 TI - Cabrol composite graft for aortic root replacement: echocardiographic imaging. PMID- 19299771 TI - Paradoxical embolization by amniotic fluid seen on the transesophageal echocardiography. PMID- 19299772 TI - Successful use of bivalirudin for combined carotid endarterectomy and coronary revascularization with the use of cardiopulmonary bypass in a patient with an elevated heparin-platelet factor 4 antibody titer. AB - We report a combined carotid endarterectomy and coronary revascularization surgery with cardiopulmonary bypass using bivalirudin for systemic anticoagulation in a patient with a positive titer for the heparin-platelet factor 4 antibody. The patient experienced procedural success for both the carotid and coronary surgeries. Increased blood and blood product transfusion was required postoperatively. PMID- 19299773 TI - Management of a patient with lupus anticoagulant and antiphospholipid syndrome for off-pump coronary artery bypass grafting using the Hepcon system. AB - Patients with serum lupus anticoagulant antibodies (LAC) with or without antiphospholipid syndrome who present for cardiac surgery provide a unique set of challenges. Chief among these are the interference with anticoagulation monitoring by LAC. We present a case of such a patient who presented to us for coronary artery bypass grafting. We follow with a review of LAC and antiphospholipid syndrome and present a strategy for ensuring adequate anticoagulation during cardiac surgery in the background of previously published reports. PMID- 19299774 TI - Cerebral oximetry during infant cardiac surgery: evaluation and relationship to early postoperative outcome. AB - BACKGROUND: We examined changes in cerebral oxygen saturation during infant heart surgery and its relationship to anatomic diagnosis and early outcome. METHODS: Regional cerebral oxygen saturation (rSO(2)) was measured by near-infrared spectroscopy in 104 infants undergoing biventricular repair without aortic arch obstruction as part of a randomized trial of hemodilution to a hematocrit of 25% vs 35%. RESULTS: Before cardiopulmonary bypass (CPB), infants with tetralogy of Fallot had higher rSO(2) values compared to those with D-transposition of the great arteries (D-TGA) or ventricular septal defect (P < 0.001). During CPB cooling, low flow, and at the termination of CPB, D-TGA subjects had the highest rSO(2) values (P < 0.001). There were no significant associations between intraoperative rSO(2) and early postoperative outcomes after adjustment for diagnosis. In 39 D-TGA subjects with > or =5 min of deep hypothermic circulatory arrest (DHCA), there was no correlation between the rSO(2) (91% +/- 6%) or hematocrit (29.2% +/- 5.5%) at the onset of arrest and the rate of decline in rSO(2) during arrest. CONCLUSIONS: Intraoperative rSO(2) varies according to anatomic diagnosis but accounts for very little of the variance in early outcome. As measured by frontal near-infrared spectroscopy, higher levels of hematocrit and current perfusion techniques appear to provide an adequate oxygen reservoir prior to relatively short periods of DHCA. PMID- 19299775 TI - Subtenon block compared to intravenous fentanyl for perioperative analgesia in pediatric cataract surgery. AB - BACKGROUND: General anesthesia with opioids provides good operative conditions for ocular surgery in children; however, postoperative pain management remains a significant problem. Regional anesthesia is commonly used as an adjunct to general anesthesia in children. We compared the efficacy and safety of subtenon block (SB) versus IV fentanyl for perioperative analgesia in pediatric cataract surgery. We hypothesized that perioperative analgesia using SB may reduce the requirement of postoperative rescue analgesia compared with fentanyl. METHODS: This was a prospective, randomized, controlled, double-blind trial. One hundred fourteen ASA I and II children (6 mo-6 yr) undergoing elective cataract surgery in one eye under general anesthesia were studied. Children were randomly allocated to one of the two groups, i.e., Group SB (n = 58) or Group F (n = 56) after securing the airway. Children in Group SB received SB with 0.06-0.08 mL/kg of 2% lidocaine and 0.5% bupivacaine (50:50) mixture and simultaneous 0.2 mL/kg normal saline IV, whereas children in Group F received 1 microg/kg (0.2 mL/kg of 5 microg/kg) of fentanyl IV and simultaneous subtenon injection with normal saline (0.06-0.08 mL/kg). Surgery started after 5 min of study drug administration. Postoperative assessment for pain, sedation, and nausea/vomiting was done at 0.5, 1, 2, 3, 4, and 24 h. The primary outcome was number of patients requiring rescue analgesia during the 24-h study period. Secondary outcomes assessed were pain and sedation scores, time to first rescue analgesia, incidence of occulocardiac reflex, and nausea/vomiting. RESULTS: The number of patients requiring rescue analgesia during the 24 h was significantly less in Group SB (n = 17/58, 29.3%) compared with Group F (n = 39/56, 69.6%, P < 0.001). The postoperative pain scores were statistically lower in Group SB at all time intervals. The median (range) time to first analgesic requirement was significantly prolonged in Group SB (9 [corrected] [2-13] vs 4 [0.5-8.5] h in Group F) (P < 0.001). Sedation scores at (1/2) h were comparable, after which significantly more children were anxious or crying in Group F compared with Group SB in which more children were calm, sitting, or lying with eyes open and relaxed (P < 0.05). A significantly higher incidence of oculocardiac reflex was recorded in Group F versus Group SB (P = 0.019). No complication related to SB was noticed. CONCLUSIONS: SB is a safe and superior alternative to IV fentanyl for perioperative analgesia in pediatric cataract surgery. PMID- 19299776 TI - The effect of pregabalin on preoperative anxiety and sedation levels: a dose ranging study. AB - BACKGROUND: Pregabalin is a gabapentinoid compound, which has been alleged to possess anxiolytic, analgesic, and anticonvulsant properties. We hypothesized that premedication with oral pregabalin would produce dose-related reductions in acute (state) anxiety and increases in sedation (sleepiness) before induction of general anesthesia. A secondary objective was to determine if premedication with pregabalin would reduce postoperative pain. METHODS: One hundred eight ASA I-III outpatients undergoing elective surgery were randomly assigned to one of the four premedication treatment groups: 1) control group received placebo capsules, 2) pregabalin 75 group received pregabalin 75 mg, po, 3) pregabalin 150 group received pregabalin 150 mg, po, and 4) pregabalin 300 group received pregabalin 300 mg, po. The effects of the study drug on the patients' level of anxiety, sedation, and pain were assessed at baseline (immediately before study drug administration), at 30 and 60 min after drug administration, and immediately before induction of anesthesia, as well as at 30-min intervals in the postanesthesia care unit (PACU) using standardized 11-point verbal rating scales, with 0 = none to 10 = maximal effect. The need for postoperative opioid analgesic medication, incidence of nausea and vomiting, requirement for rescue antiemetics, and times to discharge from the PACU and hospital, as well as the patients' quality of recovery scores, and late recovery outcomes (e.g., resumption of dietary intake and recovery of bowel function) were assessed at a 7-day follow-up interview. RESULTS: Demographic characteristics, times between study drug administration to anesthetic induction, type of surgical procedures, duration of anesthesia, PACU and hospital discharge time, as well as the requirement for fentanyl in the PACU, did not differ among the four study groups. Anxiety levels remained unchanged during the preoperative evaluation period, and did not differ among the four study groups. Sedation scores were significantly higher in the pregabalin 300 group at the preinduction assessment interval and at 90 and 120 min after surgery compared with the control group (5 +/- 3 vs 3 +/- 2, 7 +/- 4 vs 5 +/- 3, 8 +/- 4 vs 4 +/- 4, respectively, P < 0.05). CONCLUSION: Preoperative pregabalin administration (75-300 mg po) increased perioperative sedation in a dose-related fashion, but failed to reduce preoperative state anxiety, postoperative pain, or to improve the recovery process after minor elective surgery procedures. PMID- 19299777 TI - Melatonin provides anxiolysis, enhances analgesia, decreases intraocular pressure, and promotes better operating conditions during cataract surgery under topical anesthesia. AB - BACKGROUND: Melatonin has anxiolytic and potential analgesic effects. In this study, we assessed the effects of melatonin premedication on pain, anxiety, intraocular pressure (IOP), and operative conditions during cataract surgery under topical analgesia. METHODS: Forty patients undergoing cataract surgery under topical anesthesia were randomly assigned into two groups (20 patients each) to receive either melatonin 10 mg tablet (melatonin group) or placebo tablet (control group) as oral premedication 90 min before surgery. Anxiety scores, verbal pain scores, heart rate, mean arterial blood pressure, and IOP were recorded. In addition, the surgeon was asked to rate operating conditions. RESULTS: Melatonin significantly reduced the anxiety scores (median, interquartile range) from 5, 3.5-6 to 3, 2-3 after premedication and to 3, 2-3.5 during surgery (P = 0.04 and P = 0.005 compared with the placebo group, respectively). Perioperative verbal pain scores were significantly lower in the melatonin group with less intraoperative fentanyl requirement (median, interquartile range) compared with the control group, 0, 0-32.5 vs 47.5, 30-65 microg, respectively, P = 0.007. Melatonin also decreased IOP (mean +/- sd) significantly from 17.9 +/- 0.9 to 14.2 +/- 1.0 mm Hg after premedication and to 13.8 +/- 1.1 mm Hg during surgery (P < 0.001). It also provided better quality of operative conditions. CONCLUSION: We concluded that oral melatonin premedication for patients undergoing cataract surgery under topical anesthesia provided anxiolytic effects, enhanced analgesia, and decreased IOP resulting in good operating conditions. PMID- 19299778 TI - The effect of melatonin on sleep quality after laparoscopic cholecystectomy: a randomized, placebo-controlled trial. AB - BACKGROUND: In this study, we investigated whether melatonin administration could improve postoperative subjective sleep quality and reduce discomfort. METHODS: One hundred twenty-one patients scheduled for elective ambulatory laparoscopic cholecystectomy were randomized to oral 5 mg melatonin (n = 60) or placebo (n = 61) for 3 nights after surgery. Subjective sleep quality, sleep duration, sleep timing, and subjective discomfort (fatigue, general well-being, and pain) were measured. RESULTS: Sleep latency was significantly reduced in the melatonin group (mean [sd] 14 min [18]) compared with placebo (28 min [41]) on the first postoperative night (P = 0.015). The rest of the measured outcome variables did not differ between groups. CONCLUSIONS: Melatonin did not improve subjective sleep quality or discomfort compared with placebo after laparoscopic cholecystectomy. PMID- 19299779 TI - The effect of low-dose remifentanil on responses to the endotracheal tube during emergence from general anesthesia. AB - BACKGROUND: Emergence from general anesthesia can be associated with coughing, agitation, and hemodynamic disturbances. Remifentanil may attenuate these responses. METHODS: In a prospective, double-blind, randomized trial, we enrolled 60 adult patients undergoing nasal surgery using remifentanil-based anesthesia. During the emergence phase, the remifentanil group had remifentanil reduced to one tenth of the maintenance rate, whereas the control group had remifentanil discontinued. RESULTS: Times to awakening and tracheal extubation were similar between the two groups. During emergence, the remifentanil group (infusion rate 0.014 +/- 0.011 microg x kg(-1) x min(-1)) had a significantly lower incidence (40% vs 80%, P = 0.002) and less severe coughing compared with the control group, as well as a lower incidence of nonpurposeful movement (3.3% vs 30%, P = 0.006) and slower heart rates. CONCLUSIONS: Low-dose remifentanil during emergence did not prolong wake-up but reduced the incidence and severity of coughing from the endotracheal tube. PMID- 19299780 TI - Intralipid infusion diminishes return of spontaneous circulation after hypoxic cardiac arrest in rabbits. AB - BACKGROUND: Infusion of lipid emulsion has been shown to reverse lipophilic drug induced cardiovascular collapse in laboratory models and humans. The effect of high dose lipid in nondrug-induced cardiac arrest is, however, uncertain. In a rabbit model of asphyxial pulseless electrical activity (PEA) we compared lipid augmented with standard advanced cardiac life support (ACLS) resuscitation. METHOD: Adult New Zealand White rabbits underwent hypoxic PEA via tracheal clamping. After 2 min of cardiac arrest, basic life support cardiopulmonary resuscitation was commenced and 3 mL/kg 20% Intralipid or 3 mL/kg 0.9% saline solution infused. Adrenaline (100 microg/kg) was administered at 4 and 5 min. Return of spontaneous circulation (ROSC), hemodynamic metrics, and survival to 50 min were recorded. RESULTS: Seven of 11 saline-treated rabbits developed ROSC versus 1 of 12 Intralipid-treated animals; P = 0.009. No significant difference in survival to 50 min was observed (3/11 saline vs 0/12 Intralipid; P = 0.211). CONCLUSION: In this model of hypoxia-induced PEA, standard ACLS resulted in greater coronary perfusion pressure and increased ROSC compared with ACLS plus lipid infusion. Lipid emulsion may be contraindicated in cardiac arrest complicated by significant hypoxia. PMID- 19299781 TI - The counteraction of opioid-induced ventilatory depression by the serotonin 1A agonist 8-OH-DPAT does not antagonize antinociception in rats in situ and in vivo. AB - BACKGROUND: Spontaneous breathing during mechanical ventilation is gaining increasing importance during intensive care but is depressed by narcotics, such as opioids. Serotonin 1A-receptor (5-HT(1A)-R) agonists have been shown to antagonize opioid-induced ventilatory depression, but both enhancement and attenuation of nociceptive reflexes have been found with different experimental models. To clarify contradictory findings, we simultaneously determined dose response functions of the standard 5-HT(1A)-R-agonist 8-OH-DPAT and two different opioids for spontaneous ventilation and nociception. Two hypotheses were tested: 1) 8-OH-DPAT at a dose to stimulate spontaneous breathing does not activate nociceptive reflexes. 2) 8-OH-DPAT does not diminish opioid-induced antinociception. METHODS: (A) A dose-response relationship of 8-OH-DPAT, spontaneous phrenic nerve activity and a nociceptive C-fiber reflex (CFR) were established simultaneously in an in situ perfused, nonanesthetized, rat brainstem spinal cord preparation. (B) Fentanyl was administered in situ to investigate the interaction with 8-OH-DPAT on phrenic nerve activity and nociceptive CFR. Additional experiments involved the selective 5-HT(1A)-R-antagonist WAY 100 635 to exclude effects of receptors other than 5-HT(1A)-R. (C) The effects of 8-OH DPAT on spontaneous ventilation and nociceptive tail-flick reflex with and without morphine were verified in in vivo anesthetized rats. RESULTS: Low-dose 8 OH-DPAT (0.001 and 0.01 microM in situ, 0.1 microg/kg in vivo) enhanced nociceptive reflexes but did not activate spontaneous ventilation. On the contrary, high doses of 8-OH-DPAT (1 microM in situ and 10-100 microg/kg in vivo) stimulated ventilation, whereas nociceptive CFR amplitude in situ returned to baseline and tail-flick reflex was depressed in vivo. Opioid-induced ventilatory depression was antagonized by 8-OH-DPAT (1 microM in situ, and 10 microg/kg in vivo), whereas antinociception sustained. Selective 5-HT(1A)-R-antagonist WAY 100 635 (1 microM) prevented the effects of 8-OH-DPAT in situ. CONCLUSION: 5-HT(1A)-R agonist 8-OH-DPAT activates spontaneous breathing without diminishing opioid induced antinociception in rats. PMID- 19299782 TI - The effects of benzodiazepines on urotensin II-stimulated norepinephrine release from rat cerebrocortical slices. AB - BACKGROUND: Urotensin II (UII) and its receptor (UT) are implicated in mood disorders, such as stress and anxiety, and this may result, at least in part, from increased norepinephrine release from the cerebral cortex. Benzodiazepines have been widely used as hypnotics and anxiolytics, producing a decrease in cerebrocortical norepinephrine release. We hypothesized that there was some interaction between benzodiazepines and the UII system in the cerebral cortex. METHODS: In the present study, we have examined the effects of benzodiazepines on UII-increased norepinephrine release from rat cerebrocortical slices and intracellular Ca(2+) concentrations ([Ca(2+)]i) in HEK293 cells expressing rat UT receptor (HEK293-rUT cells). RESULTS: Midazolam, diazepam and flunitrazepam concentration-dependently inhibited UII-evoked norepinephrine release but did not affect [Ca(2+)]i. The IC(50) of midazolam for inhibition of UII-evoked norepinephrine release (0.32 microM, P < 0.01) was significantly lower than that of diazepam (187 microM) or flunitrazepam (40 microM). The inhibitory effects of midazolam on UII-evoked norepinephrine release were significantly attenuated by flumazenil, a benzodiazepine site antagonist. CONCLUSION: The present study suggests that midazolam, at clinically relevant concentration, significantly inhibited UII-evoked norepinephrine release. This inhibitory effect may be partially mediated via central benzodiazepine receptors. PMID- 19299783 TI - Death from propofol: accident, suicide, or murder? AB - A 24-yr-old woman was found dead in her home from apparent propofol "toxicity." Her blood level of propofol was 4.3 microg/mL. She had no history of drug abuse and no evidence of such behavior at autopsy. The medical examiner and police investigators felt that she died from probable homicide. Attention was focused on a male registered nurse acquaintance, who had acquired propofol and other drugs in the course of his regular duties in a surgical intensive care unit. This is the first reported case of murder with propofol. PMID- 19299784 TI - In vivo detection of myocardial ischemia in pigs using visible light spectroscopy. AB - BACKGROUND: Monitoring tissue oxygenation (StO(2)) by visible light spectroscopy (VLS) can identify tissue ischemia, but its feasibility for detecting myocardial ischemia is not known. We hypothesized that VLS can reliably detect changes in myocardial StO(2) in pigs subjected to acute regional or global myocardial ischemia. METHODS: In 11 pigs, regional myocardial ischemia was created by ligation of left anterior descending artery (LAD). Myocardial StO(2) was determined from the ischemic and nonischemic left ventricular (LV) regions and compared to coronary venous saturations. Myocardial function was assessed by echocardiography. In six pigs, LV-StO(2) was measured during cardiopulmonary bypass (CPB), after cardioplegic cardiac arrest, and during CPB with inadequate myocardial protection. Additionally, right ventricular (RV)- and LV-StO(2) were assessed during acute RV pressure overload from pulmonary artery (PA) banding. RESULTS: StO(2) baselines in pigs undergoing LAD occlusion were similar in the ischemic and nonischemic myocardium (70% +/- 8% vs 74% +/- 5%). After LAD ligation, StO(2) rapidly declined (30 s: 59% +/- 8%; 1 min:50 +/- 9; 5 min:42% +/ 4%; P < 0.05) in the ischemic myocardium. Decreases in StO(2) correlated with coronary venous saturations (r = 0.88) and were associated with myocardial dysfunction. In pigs undergoing CPB, LV-StO(2) remained unchanged with initiation of CPB or after cardioplegic cardiac arrest, but LV ischemia was detected by StO(2) after aortic cross-clamp without adequate myocardial protection. Similarly, PA banding resulted in a profound decrease of RV-StO(2) from 69% +/- 6% to 52% +/- 7% (P < 0.05) with recovery after PA release. CONCLUSIONS: VLS is a reliable method of detecting alterations in myocardial StO(2) and can be a useful monitor for rapid identification of myocardial ischemia. PMID- 19299785 TI - Time to a 90% change in gas concentration: a comparison of three semi-closed anesthesia breathing systems. AB - BACKGROUND: The speed with which gas concentration can be changed in the anesthesia breathing system affects the rate of denitrogenation, anesthesia induction, and emergence. Breathing system design also affects the speed at which gas concentration can be changed during maintenance. In this study, we sought to determine the speed of changes in gas concentration in modern semi-closed breathing systems. We hypothesized that equilibrium would be reached most quickly in breathing systems with smaller volume, and at high fresh gas flows. METHODS: Three anesthesia workstations were studied in vitro: the ADU (Datex-Ohmeda, now a division of GE Medical, Madison, WI), the Fabius GS with a COSY-1 breathing system (Draeger Medical, Telford, PA), and the Aestiva (Datex-Ohmeda, now a division of GE Medical, Madison, WI). The breathing systems were connected to a test lung and ventilated with air. The fresh gas flow was then changed to oxygen at rates of 1, 2, 4, 6, or 8 L/min, and times to 50%, 63%, 66%, 75%, and 90% change in oxygen concentration within the test lung were recorded. Ten trials were performed for each breathing system, at each fresh gas flow. The results were analyzed with a split-plot analysis of variance followed by post hoc tests with a Bonferroni correction. RESULTS: At flows of 6 or 8 L/min, times to equilibration did not differ among the three breathing systems. At flows of 1 to 2 L/min, the gas concentration changed faster with the ADU than with the Aestiva or Fabius (P < 0.001). At 4 L/min, the ADU was faster than Aestiva (P < 0.001), but not Fabius. CONCLUSIONS: We concluded that, other than fresh gas flow rate, breathing system volume has the biggest effect on time to equilibrium when the composition of the fresh gas inflow is changed. The position of components (e.g., valves, carbon dioxide absorber, fresh gas inlet, ventilator bellows or piston) within the breathing system has a less pronounced effect. PMID- 19299786 TI - Fluid flow through intravenous cannulae in a clinical model. AB - BACKGROUND: Predicting flow through an IV cannula is useful to clinicians if changes in flow are required and to guide selection of cannula. We sought the usefulness of manufacturers' quoted flows in predicting actual flow and to characterize that flow. METHODS: We built a vein model and inserted cannulae between 14 and 20-gauge. In the first experiment, we compared the manufacturer's quoted flows using deionized water, Hartmann's solution and Gelofusine. In the second experiment, we varied the pressure feeding the cannula and measured the resulting flow. RESULTS: Flow through a cannula is not a simple ratio of the manufacturers' quoted flow rate, even controlling for fluid type and feeding pressure. Flow is neither fully laminar, nor fully turbulent in the range of rates we have measured and in the International Organization for Standardization test. The Reynolds number is often below 2000. CONCLUSIONS: Flow through cannulae is not laminar at the upper range of clinically used flows, therefore Poiseuille's law is not useful in predicting flow and the effect of changing radius is less than commonly believed. The quoted maximum flows are also not useful. There are many conditions for laminar flow apart from Reynolds number. Further work would determine useful predictors of flow. PMID- 19299787 TI - The impact of an electronic reminder on the use of alarms after separation from cardiopulmonary bypass. AB - INTRODUCTION: During cardiopulmonary bypass (CPB) monitor alarms are routinely disabled. Failure to reactivate these alarms after CPB may jeopardize patient safety. We have produced an electronic reminder that automatically alerts clinicians to reactivate alarms after CPB and have evaluated the alarm reactivation rate after its implementation. METHODS: We developed and implemented an algorithm that identifies separation from CPB by the return of pulsatile flow and of mechanical ventilation, and checks alarm status (activated, disabled or silenced). If alarms have not been reactivated after separation from CPB, an electronic reminder appears. Data were collected during three time periods: Stage I (304 patients)--baseline period before implementation of the electronic reminder; Stage II (256 patients)--after implementation; Stage III-(435 patients) after a single educational departmental meeting, at the end of Stage II. Incidence of proper alarm reactivation and the number of electronic reminders per patient were compared among stages. RESULTS: The rate of alarm reactivation at baseline (Stage I) was 22%, increased to 63% (Stage II), and again to 83% during Stage III (P < 0.001). The spontaneous alarm reactivation rate before the appearance of the electronic reminder on the anesthesia information management system screen increased from 19% at Stage II to 42% at stage III (P < 0.001). CONCLUSION: Introducing an automatic electronic reminder significantly increased the rate of alarm reactivation after separation from CPB. Real-time computerized decision-support tools can be developed within anesthesia information management system and may be useful for improving safety during anesthesia. PMID- 19299788 TI - Error in central venous pressure measurement. AB - BACKGROUND: The variability introduced by inconsistent placement of pressure transducers for invasive monitoring may result in significant measurement error. Our goals in this study were to quantify the degree of variation among health care providers and to identify a simple tool for reducing this error. METHODS: A sample of 50 perioperative health care providers was recruited and asked to place a transducer at the appropriate level for central venous pressure (CVP) monitoring on two separate occasions: first without any additional standardization tools and second with a laser level to guide transducer placement. The variability among providers was calculated, and the results between sessions compared. RESULTS: There was significant variation in transducer placement during both sessions, in some instances, of greater magnitude than a normal CVP value. The laser level did not significantly reduce this variation. CONCLUSION: There is significant variation in transducer placement among health care providers. This variation is not reduced by a laser level and must be considered when interpreting CVP data. Hospital- or institution-wide standardization of a zero-level should be considered. PMID- 19299789 TI - Telemedicine consultation and monitoring for pediatric liver transplant. AB - Telemedicine provides the opportunity to bring medical expertise to the bedside, even if the medical expert is not physically near the patient. Internet technology has facilitated telemedicine allowing for voice, video and other data to be exchanged between remote locations. To date, applications of telemedicine to anesthesia (Teleanesthesia) have been limited. Previous work by Cone et al., (Anesth Analg 2006;1463-7) demonstrated the ability to direct an anesthetic in a remote location using satellite communication. In this report, we describe the use of telemedicine to support two cases of elective living related pediatric liver transplants performed at the Narayana Hrudayalaya Institute of Medical Sciences in Bangalore, India with preoperative and intraoperative consultation provided by physicians at the Children's Hospital of Philadelphia. PMID- 19299790 TI - A comparison of lighted stylet (Surch-Lite) and direct laryngoscopic intubation in patients with high Mallampati scores. AB - BACKGROUND: A lighted stylet is an effective alternative to a direct laryngoscope and has been reported to be particularly useful in patients with difficult airways. A high Mallampati class indicates poor visibility of the oropharyngeal structures. Because a lighted stylet does not require direct oropharyngeal visualization, we hypothesized that the lighted stylet would be easier to use than a direct laryngoscope in patients with a high Mallampati score. To examine our hypothesis, we performed a prospective, randomized study comparing a lighted stylet (Surch-Lite) with direct laryngoscopy in patients with high Mallampati scores. Success rate, time required for intubation, and hemodynamic changes were compared. METHODS: Mallampati Class III patients were enrolled and were randomly assigned to the Surch-Lite group (Group SL) or the direct laryngoscopy group (Group DL). Patients' tracheas were intubated with the randomly selected intubation device after induction of general anesthesia. Heart rate (HR) and mean arterial blood pressure were measured immediately before and every 30 s after intubation for 5 min. The time to intubation and success rate were recorded. Postoperative pharyngolaryngeal complaints were also assessed. RESULTS: Thirty patients were enrolled in each group. The success rate on the first attempt was significantly higher in Group SL (29 of 30) than in Group DL (24 of 30). The difference between maximal HR and baseline HR was significantly higher in Group DL (25 +/- 13 bpm) than in Group SL (16 +/- 10 bpm). The change in mean arterial blood pressure was also higher in Group DL (38 +/- 14 mm Hg) than in Group SL (20 +/- 13 mm Hg). The time to intubation was significantly shorter in Group SL (12 +/- 6 s) than in Group DL (17 +/- 12 s). Postoperative pharyngolaryngeal complaints were not significantly different between the two groups. CONCLUSIONS: The Surch-Lite showed a higher success rate on the first intubation attempt and produced an attenuated hemodynamic response to endotracheal intubation of patients with high Mallampati score. Thus, the Surch-Lite is an effective alternative to direct laryngoscopy in these patients. PMID- 19299791 TI - Crisis resource management of the airway in a patient with Klippel-Feil syndrome, congenital deafness, and aortic dissection. AB - Klippel-Feil syndrome is a visually arresting deformity wherein severe restriction of cervical motion predicts a difficult airway. Even minor distraction of the neck risks cervical spine or neurologic injury, so regional techniques, awake fiberoptic intubation, or awake tracheostomy are recommended anesthetic approaches. We present a case of aortic dissection in a Klippel-Feil syndrome patient for whom congenital bilateral deafness, coupled with the urgency of the surgery, mitigated against the recommended first-choice techniques. Using anesthesia crisis resource management methods, a multi-member team rehearsed predefined roles and then managed the airway via inhaled induction of anesthesia, followed by flexible fiberoptic intubation. PMID- 19299792 TI - Tracheal tube exchange: feasibility of continuous glottic viewing with advanced laryngoscopy assistance. AB - Trachea tube exchange via an airway exchange catheter is commonly combined with conventional laryngoscopy to assist intubation of the trachea. Glottic visualization may not be possible in the difficult airway. A delay in reintubation, airway injury, or intubation failure may complicate "blind" tracheal intubation because of excessive endotracheal tube size or tip impingement on airway structures. Advanced laryngoscopic techniques offering "around the corner" visualization may overcome many of the limitations of conventional laryngoscopy's "line of sight." In this data review, I examined the feasibility and usefulness of transforming a high-risk exchange from a blind procedure into one with improved glottic visualization. PMID- 19299793 TI - A systematic review of randomized controlled trials that evaluate strategies to avoid epidural vein cannulation during obstetric epidural catheter placement. AB - BACKGROUND: In this systematic review, we evaluated the evidence for seven strategies which have been proposed to minimize the incidence of epidural vein cannulation during lumbar epidural catheter placement in pregnant women. METHODS: Multiple databases were searched to identify prospective, randomized, controlled trials between December 1966 and October 2007 that evaluated methods to avoid epidural vein cannulation after lumbar epidural catheter placement in pregnant women. Published trials were evaluated using a quality assessment tool, and results were combined to evaluate efficacy to prevent epidural vein cannulation. RESULTS: Of 90 trials screened, 30 trials were included (n = 12,738 subjects). Five strategies reduce the risk of epidural vein cannulation: the lateral as opposed to sitting position (six trials, mean (sd) quality score = 35% [11%], odds ratio (OR) 0.53 [95% confidence interval (CI) 0.32-0.86]), fluid administered through the epidural needle before catheter insertion (8 trials, quality score 48% [18%], OR 0.49 [95% CI 0.25-0.97]), single rather than multiorifice catheter (5 trials, quality score 30% [6%], OR 0.64 [95% CI 0.45 0.91]), a wire-embedded polyurethane compared with polyamide epidural catheter (1 trial, 31%, plus 4 unscored abstracts, OR 0.14 [95% CI 0.06-0.30]) and catheter insertion depth < or =6 cm (2 trials, 47% [11%], OR 0.27 [95% CI 0.10-0.74]). The paramedian as opposed to midline needle approach and smaller epidural needle or catheter gauges do not reduce the risk of epidural vein cannulation. CONCLUSION: The risk of intravascular placement of a lumbar epidural catheter in pregnancy may be reduced with the lateral patient position, fluid predistension, a single orifice catheter, a wire-embedded polyurethane epidural catheter and limiting the depth of catheter insertion to 6 cm or less. In general, low manuscript quality weakens the strength of these conclusions. PMID- 19299794 TI - A comparison of epinephrine concentrations in local anesthetic solutions using a "wash" versus measured technique. AB - INTRODUCTION: Anesthesiologists often prepare epinephrine-containing local anesthetic solutions. We compared epinephrine concentrations of solutions prepared using the "wash" technique with solutions prepared using the measured technique (using an insulin syringe), and compared epinephrine concentrations among anesthesiologists. METHODS: Five anesthesiologists prepared syringes for spinal and epidural anesthesia using both techniques. Epinephrine concentrations were measured using high-performance liquid chromatography. RESULTS: Measured technique concentrations were higher than those of the wash technique for the spinal but not epidural solutions. CONCLUSIONS: Concentrations of all measured spinal solutions were higher than the target concentrations, as were concentrations of three of five measured epidural solutions. There were significant differences among anesthesiologists. PMID- 19299795 TI - Repeated episodes of respiratory distress in an obese parturient after cesarean delivery. AB - A 25-yr-old obese parturient with mild asthma underwent an uneventful spinal anesthetic for primary cesarean delivery. Within 4 h after delivery, the patient twice developed acute shortness of breath, inspiratory stridor, and hypoxemia that required intubation. A battery of blood tests revealed no evidence of an allergic reaction. She had a normal echocardiogram and chest computed tomography, but her neck computed tomography showed an enlarged left thyroid lobe asymmetrically compressing the endotracheal tube cuff. We hypothesized that, after delivery, decreased maternal vascular capacitance increased central venous pressure such that venous engorgement of an undiagnosed goiter may have caused symptomatic tracheal compression. PMID- 19299796 TI - The effect of the Operating Room Coordinator's risk appreciation on operating room efficiency. AB - BACKGROUND: The Operating Room Coordinator (ORC) is responsible for filling gaps in every operating room (OR) schedule. We have observed differences among the personalities of the four ORCs with regard to their willingness to agree to assume more risk concerning their daily planning. The hypothesis to be tested is that the relationship between the personality of each of the four ORCs and the risk an ORC is willing to take of cases running late affects OR efficiency. METHODS: In order to judge the personality of an ORC in relation to risk-taking in planning schedules, we applied the Zuckerman-Kuhlman Personality Questionnaire in our study. Seven anesthesiologists were asked to score every ORC on willingness to take risks in planning. To analyze which risk attitude creates more OR efficiency, the daily prognosis of the ORC compared with the actual OR program outcome was registered during a 5-mo period in 2006 and 2007. We analyzed whether, in the opinion of hospital management, the costs of reserving too much OR time balances with the costs of reserving too little OR time, and whether this result is consistent with the assignment of the management tasks of the ORC. RESULTS: Seven anesthesiologists classified the four ORCs into the risk-averse group (n = 2) and the nonrisk-averse group (n = 2). The Zuckerman-Kuhlman Personality Questionnaire results for risk-seeking indicate that there is a difference in risk appreciation among the different ORCs. The main finding in our study is that the nonrisk-averse ORC plans to fill the gaps in more cases in the OR program than the risk-averse ORC does. The number of extra cases performed by the nonrisk-averse ORC as compared to a risk-averse ORC is 188 in 2006 and 174 in 2007. The average end-of-program-time per OR/day for the nonrisk-averse ORC is 34 min (+/-19 min, P = 0.0085) later than for the risk-averse ORC. We find that this hospital on average reserves more OR time for procedures than is actually required. The nonrisk-averse ORC takes more advantage of that extra OR time than the risk-averse ORC does by scheduling extra cases during office hours. The success of the nonrisk-averse ORC can be linked to the fact that there is usually time available due to this over-reserving. CONCLUSIONS: The conclusion of this study is that a nonrisk-averse ORC creates significantly less unused OR capacity without a great chance of running ORs after regular working hours or canceling elective cases scheduled for surgery compared to a risk-averse ORC. PMID- 19299797 TI - Both bias and lack of knowledge influence organizational focus on first case of the day starts. AB - BACKGROUND: The economic costs of reducing first case delays are often high, because efforts need to be applied to multiple operating rooms (ORs) simultaneously. Nevertheless, delays in starting first cases of the day are a common topic in OR committee meetings. METHODS: We added three scientific questions to a 24 question online, anonymous survey performed before the implementation of a new OR information system. The 57 respondents cared sufficiently about OR management at the United States teaching hospital to complete all questions. RESULTS: The survey revealed reasons why personnel may focus on the small reductions in nonoperative time achievable by reducing tardiness in first cases of the day. (A) Respondents lacked knowledge about principles in reducing over-utilized OR time to increase OR efficiency, based on their answering the relevant question correctly at a rate no different from guessing at random. Those results differed from prior findings of responses at a rate worse than random, resulting from a bias on the day of surgery of making decisions that increase clinical work per unit time. (B) Most respondents falsely believed that a 10 min delay at the start of the day causes subsequent cases to start at least 10 min late (P < 0.0001 versus random chance). (C) Most respondents did not know that cases often take less time than scheduled (P = 0.008 versus chance). No one who demonstrated knowledge (C) about cases sometimes taking less time than scheduled applied that information to their response to (B) regarding cases starting late (P = 0.0002). CONCLUSIONS: Knowledge of OR efficiency was low among the respondents working in ORs. Nevertheless, the apparent absence of bias shows that education may influence behavior. In contrast, presence of bias on matters of tardiness of start times shows that education may be of no benefit. As the latter results match findings of previous studies of scheduling decisions, interventions to reduce patient and surgeon waiting from start times may depend principally on the application of automation to guide decision-making. PMID- 19299798 TI - Typical savings from each minute reduction in tardy first case of the day starts. AB - BACKGROUND: Analysts and clinicians sitting in operating room (OR) committee meetings cannot evaluate rapidly whether a suggested idea to reduce delays in first case of the day starts can be beneficial economically. METHODS: Three years of data were used from a six OR outpatient surgery facility. The cost reduction from reducing the tardiness of start of first cases of the day was calculated using the method of McIntosh et al. (Anesth Analg 2006;103:1499-516), limited to ORs with at least 8 h of cases and turnovers. Results were then reported per minute reduction in tardy first case of the day starts as an approximation for rapid use in meetings. RESULTS: Each 1.0 min reduction in the tardy starts of first cases of the day in ORs with more than 8 h of cases and turnovers resulted overall in 1.1 +/- 0.1 min reduction in regularly scheduled labor costs (mean +/- se). This result was close to the 1.2 min obtained using an entirely different (simulation) method performed previously for OR time reductions. Secondary analyses confirmed that assumptions were satisfied at the facility, thereby reducing the chance that results are biased. For example, the proportions of the variance in tardiness attributable to anesthesiologists and specialties were only 1% and 3%, respectively, and there were no significant differences in tardiness among the 85 anesthesiologists or 14 specialties. CONCLUSIONS: Typical savings for reducing tardiness of first case of the day starts at a surgical suite equal the product of four values: i) 1.1 min reduction in staffed OR time per 1 min reduction in tardiness, ii) estimate for reductions in tardiness (min) per OR, iii) number of ORs at the suite with more than 8 h of cases, and iv) sum of the average compensations per regularly scheduled minute for personnel in each OR. If small, the analyst and/or clinician can promptly speak up and refocus group conversation toward other potential interventions. If large, the full return on investment analysis would be performed. PMID- 19299799 TI - Cerebral oxygen saturation-time threshold for hypoxic-ischemic injury in piglets. AB - BACKGROUND: Detection of cerebral hypoxia-ischemia (H-I) and prevention of brain injury remains problematic in critically ill neonates. Near-infrared spectroscopy (NIRS), a noninvasive bedside technology could fill this role, although NIRS cerebral O(2) saturation (Sc(O2)) viability-time thresholds for brain injury have not been determined. We investigated the relationship between H-I duration at Sc(O2) 35%, a viability threshold which causes neurophysiological impairment, to neurological outcome. METHODS: Forty-six fentanyl-midazolam anesthetized piglets were equipped with NIRS and cerebral function monitor (CFM) to record Sc(O2) and electrocortical activity (ECA). After carotid occlusion, inspired O(2) was adjusted to produce H-I (Sc(O2) 35% with decreased ECA) for 1, 2, 3, 4, 6 or 8 h in different groups, followed by survival to assess neurological outcome by behavioral and histological examination. RESULTS: For H-I lasting 1 or 2 h, ECA and Sc(O2) during reperfusion rapidly returned to normal and neurological outcomes were normal. For H-I more than 2-3 h, ECA was significantly decreased and Sc(O2) was significantly increased during reperfusion, suggesting continued depression of tissue O(2) metabolism. As H-I increased beyond 2 h, the incidence of neurological injury increased linearly, approximately 15% per h. CONCLUSION: A viability-time threshold for H-I injury is Sc(O2) of 35% for 2-3 h, heralded by abnormalities in NIRS and CFM during reperfusion. These findings suggest that NIRS and CFM might be used together to predict neurological outcome, and illustrate that there is a several hour window of opportunity during H-I to prevent neurological injury. PMID- 19299800 TI - The lower limit of cerebral blood flow autoregulation is increased with elevated intracranial pressure. AB - BACKGROUND: The cerebral perfusion pressure that denotes the lower limit of cerebral blood flow autoregulation (LLA) is generally considered to be equivalent for reductions in arterial blood pressure (ABP) or increases in intracranial pressure (ICP). However, the effect of decreasing ABP at different levels of ICP has not been well studied. Our objective in the present study was to determine if the LLA during arterial hypotension was invariant with ICP. METHODS: Using continuous ventricular fluid infusion, anesthetized piglets were assigned to 1 of 3 groups: naive ICP (n = 10), moderately elevated ICP (20 mm Hg; n = 11), or severely elevated ICP (40 mm Hg; n = 9). Gradual hypotension was induced by inflation of a balloon catheter in the inferior vena cava. The LLA was determined by monitoring cortical laser-Doppler flux. RESULTS: The naive ICP group had an average CPP at the LLA (LLA(CPP)) of 29.8 mm Hg (95% CI: 26.5-33.0 mm Hg). However, the moderately elevated ICP group had a mean LLA(CPP) of 37.6 mm Hg (95% CI: 32.0-43.2 mm Hg), and the severely elevated ICP group had a mean LLA(CPP) of 51.4 mm Hg (95% CI: 41.2-61.7 mm Hg). The LLA significantly differed among groups, and the increase in LLA correlated with the increase in ICP. CONCLUSIONS: In this atraumatic, elevated ICP model in piglets, the LLA had a positive correlation with ICP, which suggests that compensating for an acute increase in ICP with an equal increase in ABP may not be sufficient to prevent cerebral ischemia. PMID- 19299801 TI - The effects of hypocapnia and the cerebral autoregulatory response on cerebrovascular resistance and apparent zero flow pressure during isoflurane anesthesia. AB - BACKGROUND: Simultaneous recordings of arterial blood pressure (ABP) and middle cerebral artery blood velocity can be used to calculate the apparent zero flow pressure (aZFP). The inverse of the slope of the pressure-velocity relationship is known as resistance area product (RAP) and is an index of cerebrovascular resistance. There is little information available regarding the effects of vasoactive drugs, arterial carbon dioxide (Paco(2)), and impaired cerebral autoregulation on aZFP and RAP during general anesthesia. During isoflurane anesthesia, we investigated the effects of hypocapnia and the effects of a phenylephrine infusion, on aZFP and RAP. METHODS: Radial ABP and transcranial Doppler middle cerebral artery blood velocity signals were recorded in 11 adults undergoing isoflurane anesthesia. A phenylephrine infusion was used to increase ABP and ventilation was adjusted to control Paco(2). Cerebral hemodynamic variables were compared at two levels of mean ABP (approximately 80 and 100 mm Hg) and at two levels of Paco(2): normocapnia (Paco(2) 38-43 mm Hg) and hypocapnia (Paco(2) 27-34 mm Hg). Two aZFP analysis methods were compared: one based on linear regression and one based on Fourier analysis of the waveforms. RESULTS: At the lower ABP, aZFP was 23 +/- 11 mm Hg and 30 +/- 13 mm Hg (mean +/- sd) with normocapnia and hypocapnia, respectively (P < 0.001) and RAP was 0.76 +/ 0.97 mm Hg x s x cm(-1) and 1.16 +/- 0.16 mm Hg x s x cm(-1) with normocapnia and hypocapnia, respectively (P < 0.001). Similar effects of hypocapnia were seen at the higher ABP. With normocapnia, isoflurane impaired cerebral autoregulation and aZFP did not change with the increase in ABP. With hypocapnia, cerebral autoregulation was not significantly impaired and increasing ABP was associated with increased aZFP (from 30 +/- 13 to 35 +/- 13 mm Hg, P < 0.01) and increased RAP (from 1.16 +/- 0.16 to 1.52 +/- 0.20 mm Hg x s x cm(-1), P < 0.001). Calculation of the relative contributions of aZFP and RAP to the cerebral hemodynamic responses indicated that changes in RAP appeared to have a greater influence than changes in aZFP. The mean difference between the two methods of determining aZFP (Fourier-regression) was 0.5 +/- 3.6 mm Hg (mean +/- 2sd). CONCLUSIONS: During isoflurane anesthesia, two interventions that increase cerebral arteriolar tone, hypocapnia and the autoregulatory response to increasing ABP, were associated with increased RAP and increased aZFP. The effect of changes in RAP appeared to be quantitatively greater than the effects of changes in aZFP. These results imply that arteriolar tone influences cerebral blood flow by controlling both resistance and effective downstream pressure. PMID- 19299802 TI - The effects of spinal anesthesia on cerebral blood flow in the very elderly. AB - BACKGROUND: Aging and disease may make elderly patients particularly susceptible to hypotension during spinal anesthesia. However, the impact of small-dose bupivacaine on cerebral hemodynamics is not known. In this study, we assessed the effects of spinal anesthesia on cerebral blood flow (CBF) in very elderly patients. METHODS: We prospectively studied 20 patients aged >75 yr who underwent open surgical repair of a hip fracture and compared them with a control group of patients younger than 60 yr. Patients were placed in the lateral decubitus position to receive spinal anesthesia at L4-5 level. Noninvasive automated arterial blood pressure, heart rate, and transcranial Doppler measurements were recorded before spinal anesthesia (baseline = T0), 5 min after the end of local anesthetic injection (T1), 10 min after spinal anesthesia (T2), and in the postanesthesia care unit (T3). RESULTS: Systolic blood pressure significantly decreased at T1 (115 +/- 25 mm Hg) and T2 (114 +/- 24 mm Hg) compared with T0 (136 +/- 21 mm Hg) in the elderly group. Systolic and diastolic velocities significantly decreased compared to baseline values (at T1, T2). Significant modifications of the pulsatility index (PI) and resistance index occurred at T2 for PI and resistance index. No patient experienced an episode of bradycardia. Heart rate variations were not significantly different compared to T0. Neither hemodynamics nor cerebral effects were observed in the control group. CONCLUSION: In summary, spinal anesthesia results in a very small but statistically significant reduction of CBF velocity in very elderly patients. PMID- 19299803 TI - General health and knee function outcomes from 7 days to 12 weeks after spinal anesthesia and multimodal analgesia for anterior cruciate ligament reconstruction. AB - BACKGROUND: We previously reported that continuous perineural femoral analgesia reduces pain with movement during the first 2 days after anterior cruciate ligament reconstruction (ACLR, n = 270), when compared with multimodal analgesia and placebo perineural femoral infusion. We now report the prospectively collected general health and knee function outcomes in the 7 days to 12 wk after surgery in these same patients. METHODS: At three points during 12 wk after ACLR surgery, patients completed the SF-36 General Health Survey, and the Knee Outcome Survey (KOS). Generalized Estimating Equations were implemented to evaluate the association between patient-reported survey outcomes and (1) preoperative baseline survey scores, (2) time after surgery, and (3) three nerve block treatment groups. RESULTS: Two hundred seventeen patients' data were complete for analysis. In univariate and multiple regression Generalized Estimating Equations models, nerve block treatment group was not associated with SF-36 and KOS scores after surgery (all with P > or = 0.05). The models showed that the physical component summary of the SF-36 (P < 0.0001) and the KOS total score (P < 0.0001) increased (improved) over time after surgery and were also influenced by baseline scores. CONCLUSIONS: After spinal anesthesia and multimodal analgesia for ACLR, the nerve block treatment group did not predict SF-36 or knee function outcomes from 7 days to 12 wk after surgery. Further research is needed to determine whether these conclusions also apply to a nonstandardized anesthetic, or one that includes general anesthesia and/or high-dose opioid analgesia. PMID- 19299804 TI - Antiinflammatory and antihyperalgesic activity of C-phycocyanin. AB - BACKGROUND: C-phycocyanin (C-PC), a biliprotein found in blue green algae, such as Spirulina platensis, is often used as a dietary nutritional supplement due to its various therapeutic values. In addition, the antiinflammatory activity of C PC partly through inhibition of proinflammatory cytokine formation, inducible nitric oxide synthase (iNOS) and cyclooxygeanase-2 (COX-2) expression has been demonstrated in many in vitro and in vivo studies. However, whether C-PC also has antihyperalgesic activity in inflammatory nociception has not been investigated. METHODS: Using a carrageenan-induced thermal hyperalgesia model, we evaluated the effect of C-PC on nociception by measuring paw withdrawal latency. To clarify the mechanisms involved, the expression of iNOS and COX-2 and the formation of nitrate and tumor necrosis factor-alpha (TNF-alpha) in the rat paw were determined. RESULTS: Pre- or posttreatment with C-PC (30 or 50 mg/kg, IP) significantly attenuated carrageenan-induced inflammatory nociception and the induction of iNOS and COX-2 at the late phase, (4 h) accompanied by an inhibition of the formation of TNF-alpha, prostaglandin E(2), nitrate and myeloperoxidase activity. CONCLUSIONS: Based on these results, it is suggested that the inhibition of NO and prostaglandin E(2) over-production through suppressing iNOS and COX-2 induction and attenuation of TNF-alpha formation and neutrophil infiltration into inflammatory sites by C-PC may contribute, at least in part, to its antihyperalgesic activity. PMID- 19299805 TI - The efficacy of the AMPA receptor antagonist NS1209 and lidocaine in nerve injury pain: a randomized, double-blind, placebo-controlled, three-way crossover study. AB - BACKGROUND: Chronic neuropathic pain is inadequately treated using current therapies, with less than half of patients achieving clinically significant pain relief (defined as more than 50% pain reduction). In this study, we evaluated the AMPA/GluR5 receptor antagonist NS1209 for efficacy, safety, and tolerability in comparison with placebo and lidocaine for the treatment of chronic neuropathic pain and allodynia in patients with peripheral nerve injury. METHODS: A randomized, double-blind, placebo-controlled, three-way crossover study was designed to recruit patients with chronic neuropathic pain for IV treatment with NS1209 (322 mg), lidocaine (5 mg/kg), and placebo. Measures of spontaneous current pain and pain evoked by brush, pinprick, cold, and heat stimulation were performed at screening and at 0, 2, 4, 6, 8, and 24 h after the start of the treatment session. RESULTS: Thirteen patients completed the study. Neither NS1209 nor lidocaine showed a statistically significant effect over placebo on the primary end-point spontaneous current pain, but both compounds exhibited a statistically significant effect on the secondary end-point pain relief of overall spontaneous pain compared with placebo. Similar to lidocaine, NS1209 was superior to placebo in alleviating some key symptoms of neuropathic pain, i.e., evoked types of pain, including mechanical and cold allodynia. CONCLUSIONS: These findings are consistent with those reported for NS1209 in other models of pain and suggest that there is a role for AMPA receptor involvement in neuropathic pain in humans. Furthermore, NS1209 was safe and well tolerated at the given doses with a safety profile similar to placebo. PMID- 19299806 TI - Health-related quality of life after tricompartment knee arthroplasty with and without an extended-duration continuous femoral nerve block: a prospective, 1 year follow-up of a randomized, triple-masked, placebo-controlled study. AB - BACKGROUND: We previously provided evidence that extending an overnight continuous femoral nerve block to 4 days after tricompartment knee arthroplasty (TKA) provides clear benefits during the perineural infusion in the immediate postoperative period. However, it remains unknown if the extended infusion improves subsequent health-related quality of life between 7 days and 12 mo. METHODS: Patients undergoing TKA received a femoral perineural infusion of ropivacaine 0.2% from surgery until the following morning, at which time patients were randomized to either continue perineural ropivacaine (n = 25) or normal saline (n = 25) in a double-masked fashion. Patients were discharged with their catheter and a portable infusion pump, and catheters were removed on postoperative day 4. Health-related quality of life was measured using the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) Index preoperatively and then at 7 days, as well as 1, 2, 3, 6, and 12 mo after surgery. The WOMAC evaluates three dimensions of health-related quality of life: pain, stiffness, and physical functional disability. For inclusion in the analysis, we required a minimum of 4 of the 6 time points, including day 7 and at least 2 of mo 3, 6, and 12. RESULTS: The two treatment groups had similar WOMAC scores for the mean area under the curve calculations (point estimate for the difference in mean area under the curve for the two groups [overnight infusion group-extended infusion group] = 1.2, 95% confidence interval: -5.6 to +8.0; P = 0.72) and at all individual time points (P > 0.05). CONCLUSIONS: We found no evidence that extending an overnight continuous femoral nerve block to 4 days improves (or worsens) subsequent health-related quality of life between 7 days and 12 mo after TKA. (ClinicalTrials.gov number, NCT00135889.). PMID- 19299807 TI - Bacterial colonization after tunneling in 402 perineural catheters: a prospective study. AB - BACKGROUND: Bacterial colonization of peripheral nerve catheters is frequent, although infection is relatively rare. With central venous catheters, the tunneling of the catheter into the subcutaneous tissue significantly decreases catheter colonization and catheter-related sepsis. We evaluated the incidence of bacterial colonization in adult patients with tunnelized perineural nerve catheters. METHODS: Peripheral nerve catheters placed under sterile conditions for postoperative analgesia were evaluated prospectively. After removal, they were analyzed for colonization. Quantitative culture was used as described by Brun-Buisson for intravascular catheters. The site of insertion was monitored daily for any signs of infection. RESULTS: Four-hundred-two patients were included in the study during a 2-yr period. The mean duration of peripheral nerve catheters was 48 h (47-50.4). Positive culture occurred in 25 catheters, indicating that the incidence of colonization was 6.22% (3.8-8.5). The microbiological analysis of the catheter tip cultures revealed coagulase-negative staphylococci in 72%. Twenty-two catheters of 25 catheters each had one microorganism, and for three catheters, two microorganisms were identified. No infection was found in any patient. CONCLUSION: The incidence of perineural catheter colonization is low with subcutaneous tunneling. Controlled randomized studies are warranted to determine whether this procedure decreases the risk for infection. PMID- 19299808 TI - The effects of thoracic epidural anesthesia on hepatic blood flow in patients under general anesthesia. AB - BACKGROUND: Hepatic hypoperfusion is regarded as an important factor in the pathophysiology of perioperative liver injury. Although epidural anesthesia (EDA) is a widely used technique, no data are available about the effects on hepatic blood flow of thoracic EDA with blockade restricted to thoracic segments in humans. METHODS: In 20 patients under general anesthesia, we assessed hepatic blood flow index in the right and middle hepatic vein by use of multiplane transesophageal echocardiography before and after induction of EDA. The epidural catheter was inserted at TH7-9, and mepivacaine 1% with a median (range) dose of 10 (8-16) mL was injected. Norepinephrine (NE) was continuously administered to patients who demonstrated a decrease in mean arterial blood pressure below 60 mm Hg after induction of EDA (EDA-NE group). The other patients did not receive any catecholamine during the study period (EDA group). A further 10 patients without EDA served as controls (control group). RESULTS: In five patients, administration of NE was necessary to avoid a decrease in mean arterial blood pressure below 60 mm Hg. Thus, the EDA-NE group consisted of five patients and the EDA group of 15. In the EDA group, EDA was associated with a median decrease in hepatic blood flow index of 24% in both hepatic veins (P < 0.01). In the EDA-NE group, all five patients showed a decrease in the blood flow index of the right (median decrease 39 [11-45] %) and middle hepatic vein (median decrease 32 [7-49] %). Patients in the control group showed a constant blood flow index in both hepatic veins. Reduction in blood flow index in the EDA group and the EDA-NE group was significant in comparison with the control group (P < 0.05). In contrast to hepatic blood flow, cardiac output was not affected by EDA. CONCLUSIONS: We conclude that, in humans, thoracic EDA is associated with a decrease in hepatic blood flow. Thoracic EDA combined with continuous infusion of NE seems to result in a further decrease in hepatic blood flow. PMID- 19299809 TI - Current threshold for nerve stimulation depends on electrical impedance of the tissue: a study of ultrasound-guided electrical nerve stimulation of the median nerve. AB - BACKGROUND: Understanding the mechanisms causing variation in current thresholds for electrical nerve stimulation may improve the safety and success rate of peripheral nerve blocks. Electrical impedance of the tissue surrounding a nerve may affect the response to nerve stimulation. In this volunteer study, we investigated the relationship between impedance and current threshold needed to obtain a neuromuscular response. METHODS: Electrical nerve stimulation and impedance measurements were performed for the median nerve in the axilla and at the elbow in 29 volunteers. The needletip was positioned at a distance of 5, 2.5, and 0 mm from the nerve as judged by ultrasound. Impulse widths of 0.1 and 0.3 ms were used for nerve stimulation. RESULTS: A significant inverse relationship between impedance and current threshold was found at the elbow, at nerve-to needle distances of 5 and 2.5 mm (P = 0.001 and P = 0.036). Impedance values were significantly lower in the axilla (mean 21.1, sd 9.7 kohm) than at the elbow (mean 36.6, sd 13.4 kohm) (P < 0.001). Conversely, current thresholds for nerve stimulation were significantly higher in the axilla than at the elbow (P < 0.001, P < 0.001, P = 0.024). A mean ratio of 1.82 was found for the measurements of current thresholds with 0.1 versus 0.3 ms impulse duration. CONCLUSIONS: Our results demonstrate an inverse relationship between impedance measurements and current thresholds and suggest that current settings used for nerve stimulation may require adjustment based on the tissue type. Further studies should be performed to investigate the clinical impact of our findings. PMID- 19299810 TI - Recurrence of cardiotoxicity after lipid rescue from bupivacaine-induced cardiac arrest. AB - Accidental intravascular administration of bupivacaine during performance of a brachial block precipitated convulsions followed by asystole. The patient was rapidly resuscitated using cardiopulmonary resuscitation, supplemented by 150 mL of 20% lipid emulsion. Nonetheless, cardiac toxicity reappeared 40 min after completion of the lipid emulsion. In the absence of further lipid emulsion, amiodarone and inotropic support were used to treat cardiotoxicity. This case suggests that local anesthetic systemic toxicity may recur after initial lipid rescue. Since recurrence of toxicity may necessitate administration of additional doses of lipid emulsion, a sufficient quantity of lipid emulsion should be available when regional anesthesia is performed. PMID- 19299811 TI - Real-time three-dimensional ultrasound-assisted axillary plexus block defines soft tissue planes. AB - Two-dimensional (2D) ultrasound is commonly used for regional block of the axillary brachial plexus. In this technical case report, we described a real-time three-dimensional (3D) ultrasound-guided axillary block. The difference between 2D and 3D ultrasound is similar to the difference between plain radiograph and computer tomography. Unlike 2D ultrasound that captures a planar image, 3D ultrasound technology acquires a 3D volume of information that enables multiple planes of view by manipulating the image without movement of the ultrasound probe. Observation of the brachial plexus in cross-section demonstrated distinct linear hyperechoic tissue structures (loose connective tissue) that initially inhibited the flow of the local anesthesia. After completion of the injection, we were able to visualize the influence of arterial pulsation on the spread of the local anesthesia. Possible advantages of this novel technology over current 2D methods are wider image volume and the capability to manipulate the planes of the image without moving the probe. PMID- 19299812 TI - Notice of retraction. PMID- 19299813 TI - The assessment of sore throat following nasotracheal intubation. PMID- 19299814 TI - Retraction notice. PMID- 19299815 TI - Forced-air warming may have benefits even in short cases. PMID- 19299816 TI - Propofol alters ketamine effect on opiate-induced hyperalgesia. PMID- 19299817 TI - Platelets and survival after liver transplantation. PMID- 19299818 TI - Thrombelastography in a patient with prolonged partial thromboplastin time undergoing a kidney transplant. PMID- 19299819 TI - Deep neck abscess and mediastinitis after laryngeal mask anesthesia. PMID- 19299820 TI - Achieving lung isolation via a tracheostomy. PMID- 19299821 TI - Team approach for laryngeal mask insertion. PMID- 19299822 TI - Transesophageal echocardiography induced airway obstruction in a patient in whom the trachea had been intubated via a LMA CTrach. PMID- 19299823 TI - The EndoFlex tube enhances navigability through the nasal cavity during nasotracheal intubation. PMID- 19299824 TI - Epidural anesthesia for cesarean delivery: failure rates. PMID- 19299825 TI - Shorter oars are more effective. AB - The purpose of this investigation was to clarify the effects of blade design and oar length on performance in rowing. Biomechanical models and equations of motion were developed to identify the main forces that affect rowing performance. In addition, the mechanical connection between the propelling blade force and the force that the rower applies on the handle was established. On this basis it was found that the blade design and oar dimensions play a significant role on the rowing performance. While rowers have found empirically that larger and/or hydrodynamically more efficient blade shapes need to be rowed with shorter oars, this article explains this tendency from a biomechanical point of view. Based on the presented evidence, it can be concluded that shorter oars will allow rowers to improve the propelling forces without increasing the handle forces. These findings explain tendencies that started with the introduction of new blade shapes in 1991. A 2 x 2 factorial ANOVA was used to test the significance of the oar shortenings that occurred with the introduction of larger blade surfaces while international record times improved during all those years. Consequently, the findings of this investigation encourage coaches to further experiment with shorter oars and oar manufacturers to continue their blade development that would lead to even shorter oars, with the goal of continuous rowing performance improvements. PMID- 19299826 TI - Comparison of selected lateral cutting activities used to assess ACL injury risk. AB - The purpose of this study was to compare the knee joint dynamics for males and females performing constrained cutting tasks to an unanticipated running and cutting maneuver. Sixteen male and 17 female recreational athletes were recruited to perform four cutting tasks; unanticipated run and cut (CUT), stride land and cut (SLC), far box-land and cut (FLC), and close box-land and cut (CLC). Three dimensional knee joint kinematics and kinetics were recorded. Data were compared across gender and task with a 2 x 4 ANOVA (p < .05), and a factor analysis was performed to examine task relationships. There were significant group mean differences between the tasks and across genders. The factor analysis revealed high correlations between the three constrained tasks, but for variables typically associated with ACL injury risk there was a poor relationship to the CUT task. This was particularly true for the frontal plane variables. The constrained tasks were only moderately useful in predicting cutting mechanics. PMID- 19299827 TI - Biomechanical comparison between elite female and male baseball pitchers. AB - The purpose of the current study was to identify the biomechanical features of elite female baseball pitching. Kinematics and kinetics of eleven elite female baseball pitchers were reported and compared with eleven elite male pitchers. Results suggested that females share many similarities with males in pitching kinematics, with a few significant differences. Specifically, at the instant of stride foot contact, a female pitcher had a shorter and more open stride and less separation between pelvis orientation and upper torso orientation. From foot contact to ball release, a female pitcher produced lower peak angular velocity for throwing elbow extension and stride knee extension. Ball velocity was lower for the female. Foot contact to ball release took more time for a female pitcher. Maximal proximal forces at the shoulder and elbow joints were less for a female pitcher. PMID- 19299828 TI - Effects of speed and visual-target distance on toe trajectory during the swing phase of treadmill walking. AB - Gait kinematics have been shown to vary with speed and visual-target fixation distance, but their combined effects on toe trajectory during treadmill walking are not known. The purpose of this investigation was to determine the role of walking speed and target distance on vertical toe trajectory during treadmill walking. Subjects walked on a treadmill at five speeds while performing a dynamic visual-acuity task at both "far" and "near" target distances (ten trials total). The analysis concentrated on three specific toe trajectory events during swing: the first peak toe height just after toe-off; the minimum toe height (toe clearance), and the second peak toe height just before heel strike. With increasing speed, toe clearance decreased and the peak toe height just before heel strike increased. Only the peak toe height just after toe-off was significantly changed between the near-target and far-target tasks, though the difference was small. Therefore, walking speed and visual-fixation distance cannot be neglected in the analysis of toe trajectory. Otherwise, differences observed between populations may be attributed to age- or clinically related factors, instead of disparities of speed or target-fixation distance. PMID- 19299829 TI - A comparison between two models of shoulder muscle force estimation. AB - Shoulder injuries are often related to rotator cuff muscles. Although there are various models for muscle force estimation, it is difficult to ensure that the results obtained with such models are reliable. The aim of the current study was to compare two models of muscle force estimation. Eight subjects, seven male and one female (mean age of 24 yr; mean height of 1.83 m), performed five isokinetic maximum concentric contractions of internal and external shoulder rotation. Two models with different algorithms were used. In both, the input data consisted of the measured internal rotation moment. Comparisons were made between the difference and the average results obtained with each model of muscle force estimation. There was reasonable agreement among the results for force between the two models for subscapularis, pectoralis major, and anterior deltoideus muscles results. Conversely, poor correlation was found for the latissimus dorsi, teres major, and middle deltoid. These results suggest that the algorithm structure might have a strong effect on muscle force estimation results. PMID- 19299830 TI - Influence of running shoe midsole composition and custom foot orthotic intervention on lower extremity dynamics during running. AB - The purpose of this study was to analyze the influence of varying running shoe midsole composition on lower extremity dynamics with and without a custom foot orthotic intervention. Three-dimensional dynamics were collected on 12 female runners who had completed 6 weeks of custom foot orthotic therapy. Participants completed running trials in 3 running shoe midsole conditions-with and without a custom foot orthotic intervention. Results from the current study revealed that only maximum rearfoot eversion velocity was influenced by the midsole durometer of the shoe. Maximum rearfoot eversion velocity was significantly decreased for the hard shoe compared with the soft shoe. However, the orthotic intervention in the footwear led to significant decreases in several dynamic variables. The results suggest that the major component influencing the rearfoot dynamics was the orthotic device and not the shoe composition. In addition, data suggest that the foot orthoses appear to compensate for the lesser shoe stability enabling it to function in a way similar to that of a shoe of greater stability. PMID- 19299831 TI - Material property sensitivity analysis on resonant frequency characteristics of the human spine. AB - The aim of this study is to investigate the effect of material property changes in the spinal components on the resonant frequency characteristics of the human spine. Several investigations have reported the material property sensitivity of human spine under static loading conditions, but less research has been devoted to the material property sensitivity of spinal biomechanical characteristics under a vibration environment. A detailed three-dimensional finite element model of the human spine, T12-pelvis, was built and used to predict the influence of material property variation on the resonant frequencies of the human spine. The simulation results reveal that material properties of spinal components have obvious influences on the dynamic characteristics of the spine. The annulus ground substance is the dominant component affecting the vertical resonant frequencies of the spine. The percentage change of the resonant frequency relative to the basic condition was more than 20% if Young's modulus of disc annulus is less than 1.5 MPa. The vertical resonant frequency may also decrease if Poisson's ratio of nucleus pulposus of intervertebral disc decreases. PMID- 19299832 TI - Torso and hip muscle activity and resulting spine load and stability while using the ProFitter 3-D Cross Trainer. AB - The ProFitter 3-D Cross Trainer is a labile surface device used in the clinic and claimed to train spine stability. The purpose of this study was to quantify the spine mechanics (compression and shear forces and stability), together with muscle activation mechanics (surface electromyography) of the torso and hip, during three ProFitter exercises. Trunk muscle activity was relatively low while exercising on the device (<25%MVC). Gluteus medius activity was phasic with the horizontal sliding position, especially for an experienced participant. Sufficient spinal stability was achieved in all three exercise conditions. Peak spinal compression values were below 3400 N (maximum 3188 N) and peak shear values were correspondingly low (under 500 N). The exercises challenge whole-body dynamic balance while producing very conservative spine loads. The motion simultaneously integrates hip and torso muscles in a way that appears to ensure stabilizing motor patterns in the spine. This information will assist with clinical decision making about the utility of the device and exercise technique in rehabilitation and training programs. PMID- 19299833 TI - The muscle force component in pedaling retains constant direction across pedaling rates. AB - Changes in pedaling rate during cycling have been found to alter the pedal forces. Especially, the force effectiveness is reduced when pedaling rate is elevated. However, previous findings related to the muscular force component indicate strong preferences for certain force directions. Furthermore, inertial forces (due to limb inertia) generated at the pedal increase with elevated pedaling rate. It is not known how pedaling rate alters the inertia component and subsequently force effectiveness. With this in mind, we studied the effect of pedal rate on the direction of the muscle component, quantified with force effectiveness. Cycle kinetics were recorded for ten male competitive cyclists at five cadences (60-100 rpm) during unloaded cycling (to measure inertia) and at a submaximal load (~260 W). The force effectiveness decreased as a response to increased pedaling rate, but subtracting inertia eliminated this effect. This indicates consistent direction of the muscle component of the foot force. PMID- 19299834 TI - Feasibility of using magnetic resonance elastography to study the effect of aging on shear modulus of skeletal muscle. AB - A common complication associated with aging is the stiffening of skeletal muscles. The purpose of this study was to determine the ability of magnetic resonance elastography (MRE) to study this phenomenon in vivo. Twenty female subjects were included in the study with an age range of 50 to 70 years. Shear modulus was calculated for the tibialis anterior of each subject. There was not a significant relationship between age and shear modulus. However, three subjects had abnormally high values and were among the oldest subjects tested. There was a significant relationship between age and tissue stiffness homogeneity. More research is needed to determine whether the changes seen here are reflective of increased tissue cross-linking or related to reduced muscle quality. However, MRE shows promise as a tool to study aging-related muscle stiffness changes or to evaluate treatments to counteract these changes. PMID- 19299836 TI - Accelerometer assessment of physical activity in active, healthy older adults. AB - Despite widespread use of accelerometers to objectively monitor physical activity among adults and youth, little attention has been given to older populations. The purpose of this study was to define an accelerometer-count cut point for a group of older adults and to then assess the group's physical activity for 7 days. Participants (N = 38, age 69.7 +/- 3.5 yr) completed a laboratory-based calibration with an Actigraph 7164 accelerometer. The cut point defining moderate to vigorous physical activity (MVPA) was 1,041 counts/min. On average, participants obtained 68 min of MVPA per day, although more than 65% of this occurred as sporadic activity. Longer bouts of activity occurred in the morning (6 a.m. to 12 p.m.) more frequently than other times of the day. Almost 14 hr/day were spent in light-intensity activity. This study demonstrates the rich information that accelerometers provide about older adult activity patterns information that might further our understanding of the relationship between physical activity and healthy aging. PMID- 19299835 TI - Exercise self-efficacy and control beliefs: effects on exercise behavior after an exercise intervention for older adults. AB - The current study examined exercise self-efficacy and exercise behavior during and after a strength-training intervention program with older adults. A model with cross-lagged and contemporaneous paths was tested with structural equations. Within testing occasions, higher physical resistance was related to greater beliefs in efficacy and control over exercise. At 3 months into the intervention, those who had higher physical resistance were less likely to show subsequent changes in beliefs. Those who had higher self-efficacy and control beliefs at 6 months were more likely to report that they were still exercising at 9 and 12 months after the intervention. Findings indicate that exercise self-efficacy and exercise behavior are associated with one another and that beliefs developed during an intervention are important for maintenance of an exercise regimen. PMID- 19299837 TI - Vitamin-d-receptor genotypes and bone-mineral density in postmenopausal women: interaction with physical activity. AB - The current study investigated the association between vitamin-D-receptor (VDR) genotypes with bone-mineral density (BMD) and its interaction with physical activity level (PAL). Individuals in a sample of 192 volunteers (67.84 +/- 5.23 years) underwent BMD evaluation and were genotyped for VDR ApaI, BsmI, FokI, and TaqI polymorphisms. Haplotypes were reconstructed through expectation maximization algorithm, and regression-based haplotype-specific association tests were performed with studied phenotypes. None of the polymorphisms were associated with BMD at any site; however, haplotype was associated with femoral-neck and Ward's-triangle BMD. Interaction between PAL and VDR genotypes was significant for the FokI polymorphism at femoral-neck and Ward's-triangle BMD. The FokI T/T genotype was associated with higher BMD in active women. It was concluded that VDR haplotypes, but not genotypes, are associated with femoral-neck and Ward's triangle BMD in postmenopausal women. Moreover, the results suggest that VDR FokI polymorphism might be a potential determinant of BMD response to physical activity. PMID- 19299838 TI - Ambulatory physical activity profiles of older adults. AB - This study examined objectively determined walking profiles of older adults across a wide range of sociocultural backgrounds. All individuals (N = 415; 131 men age 70.5 +/- 9.2 yr and 284 women age 71.5 +/- 9.0 yr) underwent physiological measurements, completed pen-and-paper surveys, and wore a pedometer for 7 consecutive days. The total sample accumulated a mean of 3,987 +/-2,680 steps/day. Age (r = -.485, p < .001) and body-mass index (BMI; r = -.353, p < .001) were negatively associated with steps per day. Multivariate analysis revealed that race/ethnic category (F = 3.15, df = 3), gender (F = 2.46, df = 1), BMI (F = 6.23, df = 2), income (F = 9.86, df = 1), education (F = 43.3, df = 1), and retirement status (F = 52.3, df = 1) were significantly associated with steps per day. Collectively these categories accounted for 56% of the variance in walking activity in this independently living, community-dwelling older adult sample. Sedentary characteristics highlighted within, and step-per-day values specific to, older adults have implications for planning targeted physical activity interventions related to walking activity in this population. PMID- 19299839 TI - Getting Grounded Gracefully: effectiveness and acceptability of Feldenkrais in improving balance. AB - The Getting Grounded Gracefully program, based on the Awareness Through Movement lessons of the Feldenkrais method, was designed to improve balance and function in older people. Fifty-five participants (mean age 75, 85% women) were randomized to an intervention (twice-weekly group classes over 8 wk) or a control group (continued with their usual activity) after being assessed at baseline and then reassessed 8 wk later. Significant improvement was identified for the intervention group relative to the control group using ANOVA between-groups repeated-measures analysis for the Modified Falls Efficacy Scale score (p = .003) and gait speed (p = .028), and a strong trend was evident in the timed up-and-go (p = .056). High class attendance (88%) and survey feedback indicate that the program was viewed positively by participants and might therefore be acceptable to other older people. Further investigation of the Getting Grounded Gracefully program is warranted. PMID- 19299840 TI - Altered DHEA and DHEAS response to exercise in healthy older adults. AB - Dehydroepiandrosterone (DHEA) and its sulfate (DHEAS) are hormones produced by the adrenal cortex that decline in concentration with age. Decreased DHEA levels are associated with age-related disease and oxidative stress but might be increased in younger adults by exercise. Studies are presented assessing the response of DHEA and DHEAS to varied-intensity exercise in older age. DHEA increased significantly in young adults (14.5 +/- 6.1 ng/ml rising to 21.1 +/- 7.5 ng/ml; p < .01), whereas DHEAS decreased significantly (2.56 +/- 1.11 microg/ml falling to 1.90 +/- 0.8 microg/ml; p < .05), after submaximal exercise. DHEA and DHEAS levels were significantly lower in older adults than in younger adults (p < .01), and there was no observed response of either hormone to exercise in older adults. Lipoprotein protein carbonylation is presented as a measure of oxidative status and significantly decreased in younger adults postexercise. Participants with higher DHEA postexercise had lower LDL protein carbonyl concentrations (Pearson's coefficient -.409, p < .05). PMID- 19299841 TI - Fatigue after physical activity in healthy and balance-impaired elderly. AB - This study aimed to determine whether physical activity similar to daily living results in signs and sensations of fatigue in healthy old, balance-impaired old, and healthy young adults. Sensations of general tiredness, leg tiredness, knee extension and hip-abduction strength, and temporospatial gait variables were measured before, immediately after, and up to 20 min after moderate-intensity physical activity. After activity, all groups reported increased levels of tiredness but showed no changes in strength. The balance-impaired had greater and more prolonged feelings of tiredness, with a mean increase before to immediately after activity of 3.6 on the visual analogue scale and no recovery at 16 min. The young and healthy old had an increase of 3 and 2.6, respectively, and had recovered before 16 min. In the balance-impaired group only, cadence slowed immediately after activity. It is proposed that these changes, in particular the prolonged feelings of tiredness, might limit daily activity. PMID- 19299842 TI - Living with fear: the lived experience of community mobility among older adults with low vision. AB - This secondary analysis of data drawn from a descriptive phenomenological study explored how older adults with low vision experience and manage community mobility. Participants included 34 urban and rural older adults, age 70 years and older, who were not using low-vision-rehabilitation services. The findings convey a core element of the experience of community mobility for participants: living with a pervasive sense of fear regarding one's body and way of being. Participants continually gauged risks associated with mobility and engaged in risk avoidance and management strategies. Community mobility was often restricted by participants because of perceived risks, leading to reduced participation in a range of physical, social, and other types of activities. Further research on environmental factors mediating community mobility and on strategies effective in maintaining mobility among seniors with low vision is essential to optimize participation, health, and service delivery. PMID- 19299843 TI - Fall risk factors in older female lawn bowls players and controls. AB - Fall-risk factors were compared between older women who regularly participate in lawn bowls and controls. Participants were 74 healthy women age 60-75 yr. Postural stability, timed up-and-go score, isometric knee-extensor strength, reaction time, range of motion, and calcaneal broadband ultrasonic attenuation (BUA) were assessed. Bowlers and controls were similar in regard to mean age, height, and weight. Regular bowlers had significantly lower timed up-and-go times, reaction times, and sway while standing on a compliant surface. They had significantly higher knee-extensor strength, range of motion at ankle and shoulder, and calcaneal BUA. Although targeted strength and balance training might be necessary in treatment of the physical frailty that can contribute to falls and fractures in older people, it is possible that long-term participation in activity such as lawn bowls could help prevent this frailty. PMID- 19299844 TI - Beyond the evidence of guidelines. PMID- 19299845 TI - Microscopic colitis: an unfamiliar but treatable disease. AB - Chronic diarrhoea is a frequent complaint in clinical practice. Microscopic colitis is the cause of this symptom in 10% of these cases and the prevalence is rising. To exclude microscopic colitis a colonoscopy with multiple biopsies of different regions of the colon is mandatory. A sigmoidoscopy alone is insufficient. Two histopathological types of microscopic colitis can be distinguished: collagenous colitis and lymphocytic colitis. Nowadays, there is sufficient evidence to recommend budesonide as the first-choice treatment. Bismuth can also be recommended, but this drug is not easily available in the Netherlands. Evidence of efficacy of other drugs is scant. PMID- 19299846 TI - Vascular liver disorders (II): portal vein thrombosis. AB - Portal vein thrombosis (PVT) is a rare disorder that is associated with a variety of underlying conditions, of which liver cirrhosis, malignancy and myeloproliferative disorders are the most common. Based on clinical presentation and results of imaging, two different entities can be identified, acute and chronic PVT. Anticoagulation therapy is recommended for all patients with acute PVT in an attempt to prevent further thrombosis and to promote recanalisation of the obstructed veins. Chronic PVT is characterised by the presence of a portal cavernoma and development of portal hypertension. Bleeding from ruptured oesophageal or gastric varices is the main complication of portal hypertension in these patients. Both endoscopic therapy and beta-adrenergic blockade are used for the prevention and treatment of gastrointestinal bleeding. In the absence of bleeding, continuous anticoagulant therapy should be considered for the group of chronic PVT patients in whom an underlying prothrombotic factor can be identified. With adequate management of complications and concurrent diseases, prognosis of PVT is good in patients without underlying cirrhosis or malignancies. PMID- 19299847 TI - Urinary excretion of low-molecular-weight proteins as prognostic markers in IgA nephropathy. AB - BACKGROUND: Immunoglobulin A nephropathy (IgAN) is characterised by high variability in clinical course and outcome. Accurate prediction of prognosis is needed to optimise treatment. Urinary alpha1-microglobulin and beta2 microglobulin are markers of tubulointerstitial injury and predict the risk of end-stage renal disease (ESRD) in idiopathic membranous nephropathy. We questioned the relevance of these markers in IgAN. METHODS: We included patients with biopsy proven IgAN, who were evaluated for proteinuria in our centre between 1995 and 2007. Data were analysed using univariate and multivariate Cox regression for the outcome variables ESRD and progression (rise in serum creatinine of >50% or start of immunosuppressive therapy). RESULTS: Seventy patients (71% men) were selected. Median age was 39 years, median serum creatinine 140 micromol/l, and median proteinuria 2.4 g/day. Median urinary alpha1-microglobulin excretion was 23.5 microg/min (range 3.5-275.3) and median urinary beta2-microglobulin excretion was 0.4 microg/min (range 0.1-62.1). Both alpha1m and beta2m correlated significantly with serum creatinine (r = 0.65, p<0.01 and r = 0.62, p<0.01) and total proteinuria (r = 0.35, p<0.01 and r = 0.28, p<0.05). During follow-up (median 75 months) 25 patients (36%) developed ESRD , and 46 patients (66%) showed progression. 19 patients (27%) were treated with immunosuppressive agents. In univariate analysis urinary alpha1- and beta2 microglobulin predicted ESRD and progression. In multivariate analysis only serum creatinine and urinary protein were independent predictors of both outcomes. CONCLUSION: Urinary excretion of low molecular weight proteins did not offer an advantage over total proteinuria and serum creatinine in predicting prognosis in patients with IgAN. PMID- 19299848 TI - Value judgements that matter to patients remain implicit in oncology guidelines: an observational study. AB - BACKGROUND: Clinical practice guidelines are often evidence-based. However, it is inevitable that there are value judgements in the practical recommendations contained in the guidelines. In order to see if patients are ultimately being supplied with sufficient information to help them make treatment decision, we determined 1) which value judgements influence the process of developing guidelines for palliative chemotherapy, and 2) whether these value judgements were made explicit in the final guideline report. METHODS: We studied the development process of six Dutch oncology guidelines in which palliative chemotherapy plays a substantial role. We observed the guideline development groups (GDGs), conducted semi-structured interviews with individual GDG members (including the chairs), and analysed the minutes of GDG meetings and subsequent versions of the guidelines. A value judgement was defined as a statement about the value of a patient outcome with regard to palliative chemotherapy. RESULTS: We identified the following value judgements in the process of guideline development: 1) consensus on what should be considered as valuable minimum patient outcomes, 2) preference for tailored treatment in situations where there is no evidence of treatment effect, 3) preference for 'doing something' even when there is sufficient evidence of no effect, and 4) the patient outcome of 'prolonging life'. These value judgements, however, were not reported in the final guideline. CONCLUSION: At least the last two value judgements mentioned are relevant for patients with incurable metastatic cancer in making decisions whether to undergo chemotherapy and what kind. Value judgements should be made explicit in guidelines, so that clinicians can transparently discuss treatment options with individual patients. PMID- 19299849 TI - Pylephlebitis after a duodenal ulcer in a patient with metastasised colon carcinoma treated with chemotherapy and bevacizumab: a case report. AB - Pylephlebitis or septic thrombophlebitis of the portal vein is a rare entity with a high mortality rate. It is often a complication of intra-abdominal infection most commonly caused by diverticulitis and appendicitis. Diagnosis is often delayed since clinical signs and symptoms are nonspecific. Pylephlebitis should be considered in patients with sepsis due to gut-associated organisms without a clear focus of infection. We describe a patient with metastastatic colon carcinoma treated with chemotherapy and bevacizumab who was diagnosed with pylephlebitis after a duodenal ulcer and responded well to antibiotic treatment. PMID- 19299850 TI - Ceftriaxone-induced acute reversible encephalopathy in a patient treated for a urinary tract infection. AB - Encephalopathy is a rare side effect of third- and fourth-generation cephalosporins. Renal failure and previous disease of the central nervous system predispose to this neurotoxicity. We describe a case of encephalopathy with generalised triphasic waves in a patient with pre-existent cerebrovascular disease who was treated with ceftriaxone for a urinary tract infection. Early recognition of this complication is relevant given that ceftriaxone discontinuation reverted the neurological syndrome. PMID- 19299851 TI - Investigating unexpected INRs: in search of the culprit--adherence, interactions, genetics, and superwarfarin. AB - Treatment with coumarin derivatives is highly individualised due to high intra- and inter-individual variation in dose response and risks of severe bleeding or thromboembolic complications. Treatment focuses on reaching and maintaining a stable target international normalised ratio (INR). However, unexpected INRs that are not explained by noncompliance or vitamin K intake may occur. Here we describe seven cases of unexpected INRs, and provide clues that clarify the underlying mechanism. PMID- 19299852 TI - Fever, diffuse rash and arthralgia. PMID- 19299853 TI - A patient with acute abdominal pain, elevated lactate dehydrogenase and a normal spiral computed tomography. PMID- 19299854 TI - Approximate matching of digital point sets using a novel angular tree. AB - Matching and analysis of patterns or shapes in the digital plane are of utmost importance in various problems of computer vision and pattern recognition. A digital point set is such a pattern that corresponds to an object in the digital plane. Although there exist several data structures that can be employed for Approximate Point Set Pattern Matching (APSPM) in the real domain, they require substantial modification to support algorithms in the digital domain. To bridge this gap, a novel data structure called "angular tree" is proposed, targeting an efficient and error-controllable circular range query in the digital plane. The farthest pair of points may be used as the starting correspondence between the pattern set and the background set. Several classical discrete structures and methodologies of computational geometry, as well as some topological features of circles/discs in digital geometry, have been used in tandem, for successful realization of the proposed APSPM algorithm in the digital plane. The APSPM algorithm based on the angular tree has been implemented and tested on various point sets and the reported results demonstrate the efficiency and versatility of the new data structure for supporting APSPM algorithms. PMID- 19299855 TI - Branch-and-bound methods for euclidean registration problems. AB - In this paper, we propose a practical and efficient method for finding the globally optimal solution to the problem of determining the pose of an object. We present a framework that allows us to use point-to-point, point-to-line, and point-to-plane correspondences for solving various types of pose and registration problems involving euclidean (or similarity) transformations. Traditional methods such as the iterative closest point algorithm or bundle adjustment methods for camera pose may get trapped in local minima due to the nonconvexity of the corresponding optimization problem. Our approach of solving the mathematical optimization problems guarantees global optimality. The optimization scheme is based on ideas from global optimization theory, in particular convex underestimators in combination with branch-and-bound methods. We provide a provably optimal algorithm and demonstrate good performance on both synthetic and real data. We also give examples of where traditional methods fail due to the local minima problem. PMID- 19299856 TI - Distribution-based dimensionality reduction applied to articulated motion recognition. AB - Some articulated motion representations rely on frame-wise abstractions of the statistical distribution of low-level features such as orientation, color, or relational distributions. As configuration among parts changes with articulated motion, the distribution changes, tracing a trajectory in the latent space of distributions, which we call the configuration space. These trajectories can then be used for recognition using standard techniques such as dynamic time warping. The core theory in this paper concerns embedding the frame-wise distributions, which can be looked upon as probability functions, into a low-dimensional space so that we can estimate various meaningful probabilistic distances such as the Chernoff, Bhattacharya, Matusita, Kullback-Leibler (KL) or symmetric-KL distances based on dot products between points in this space. Apart from computational advantages, this representation also affords speed-normalized matching of motion signatures. Speed normalized representations can be formed by interpolating the configuration trajectories along their arc lengths, without using any knowledge of the temporal scale variations between the sequences. We experiment with five different probabilistic distance measures and show the usefulness of the representation in three different contexts-sign recognition (with large number of possible classes), gesture recognition (with person variations), and classification of human-human interaction sequences (with segmentation problems). We find the importance of using the right distance measure for each situation. The low-dimensional embedding makes matching two to three times faster, while achieving recognition accuracies that are close to those obtained without using a low-dimensional embedding. We also empirically establish the robustness of the representation with respect to low-level parameters, embedding parameters, and temporal-scale parameters. PMID- 19299857 TI - Image transformations and blurring. AB - Since cameras blur the incoming light during measurement, different images of the same surface do not contain the same information about that surface. Thus, in general, corresponding points in multiple views of a scene have different image intensities. While multiple-view geometry constrains the locations of corresponding points, it does not give relationships between the signals at corresponding locations. This paper offers an elementary treatment of these relationships. We first develop the notion of "ideal" and "real" images, corresponding to, respectively, the raw incoming light and the measured signal. This framework separates the filtering and geometric aspects of imaging. We then consider how to synthesize one view of a surface from another; if the transformation between the two views is affine, it emerges that this is possible if and only if the singular values of the affine matrix are positive. Next, we consider how to combine the information in several views of a surface into a single output image. By developing a new tool called "frequency segmentation," we show how this can be done despite not knowing the blurring kernel. PMID- 19299858 TI - Make3D: learning 3D scene structure from a single still image. AB - We consider the problem of estimating detailed 3D structure from a single still image of an unstructured environment. Our goal is to create 3D models that are both quantitatively accurate as well as visually pleasing. For each small homogeneous patch in the image, we use a Markov Random Field (MRF) to infer a set of "plane parameters" that capture both the 3D location and 3D orientation of the patch. The MRF, trained via supervised learning, models both image depth cues as well as the relationships between different parts of the image. Other than assuming that the environment is made up of a number of small planes, our model makes no explicit assumptions about the structure of the scene; this enables the algorithm to capture much more detailed 3D structure than does prior art and also give a much richer experience in the 3D flythroughs created using image-based rendering, even for scenes with significant nonvertical structure. Using this approach, we have created qualitatively correct 3D models for 64.9 percent of 588 images downloaded from the Internet. We have also extended our model to produce large-scale 3D models from a few images. PMID- 19299859 TI - Low-rank matrix fitting based on subspace perturbation analysis with applications to structure from motion. AB - The task of finding a low-rank (r) matrix that best fits an original data matrix of higher rank is a recurring problem in science and engineering. The problem becomes especially difficult when the original data matrix has some missing entries and contains an unknown additive noise term in the remaining elements. The former problem can be solved by concatenating a set of r-column matrices that share a common single r-dimensional solution space. Unfortunately, the number of possible submatrices is generally very large and, hence, the results obtained with one set of r-column matrices will generally be different from that captured by a different set. Ideally, we would like to find that solution that is least affected by noise. This requires that we determine which of the r-column matrices (i.e., which of the original feature points) are less influenced by the unknown noise term. This paper presents a criterion to successfully carry out such a selection. Our key result is to formally prove that the more distinct the r vectors of the r-column matrices are, the less they are swayed by noise. This key result is then combined with the use of a noise model to derive an upper bound for the effect that noise and occlusions have on each of the r-column matrices. It is shown how this criterion can be effectively used to recover the noise-free matrix of rank r. Finally, we derive the affine and projective structure-from motion (SFM) algorithms using the proposed criterion. Extensive validation on synthetic and real data sets shows the superiority of the proposed approach over the state of the art. PMID- 19299860 TI - A novel connectionist system for unconstrained handwriting recognition. AB - Recognizing lines of unconstrained handwritten text is a challenging task. The difficulty of segmenting cursive or overlapping characters, combined with the need to exploit surrounding context, has led to low recognition rates for even the best current recognizers. Most recent progress in the field has been made either through improved preprocessing or through advances in language modeling. Relatively little work has been done on the basic recognition algorithms. Indeed, most systems rely on the same hidden Markov models that have been used for decades in speech and handwriting recognition, despite their well-known shortcomings. This paper proposes an alternative approach based on a novel type of recurrent neural network, specifically designed for sequence labeling tasks where the data is hard to segment and contains long-range bidirectional interdependencies. In experiments on two large unconstrained handwriting databases, our approach achieves word recognition accuracies of 79.7 percent on online data and 74.1 percent on offline data, significantly outperforming a state of-the-art HMM-based system. In addition, we demonstrate the network's robustness to lexicon size, measure the individual influence of its hidden layers, and analyze its use of context. Last, we provide an in-depth discussion of the differences between the network and HMMs, suggesting reasons for the network's superior performance. PMID- 19299861 TI - NV-tree: an efficient disk-based index for approximate search in very large high dimensional collections. AB - Over the last two decades, much research effort has been spent on nearest neighbor search in high-dimensional data sets. Most of the approaches published thus far have, however, only been tested on rather small collections. When large collections have been considered, high-performance environments have been used, in particular systems with a large main memory. Accessing data on disk has largely been avoided because disk operations are considered to be too slow. It has been shown, however, that using large amounts of memory is generally not an economic choice. Therefore, we propose the NV-tree, which is a very efficient disk-based data structure that can give good approximate answers to nearest neighbor queries with a single disk operation, even for very large collections of high-dimensional data. Using a single NV-tree, the returned results have high recall but contain a number of false positives. By combining two or three NV trees, most of those false positives can be avoided while retaining the high recall. Finally, we compare the NV-tree to Locality Sensitive Hashing, a popular method for epsilon-distance search. We show that they return results of similar quality, but the NV-tree uses many fewer disk reads. PMID- 19299862 TI - Robust estimation of albedo for illumination-invariant matching and shape recovery. AB - We present a nonstationary stochastic filtering framework for the task of albedo estimation from a single image. There are several approaches in the literature for albedo estimation, but few include the errors in estimates of surface normals and light source direction to improve the albedo estimate. The proposed approach effectively utilizes the error statistics of surface normals and illumination direction for robust estimation of albedo, for images illuminated by single and multiple light sources. The albedo estimate obtained is subsequently used to generate albedo-free normalized images for recovering the shape of an object. Traditional Shape-from-Shading (SFS) approaches often assume constant/piecewise constant albedo and known light source direction to recover the underlying shape. Using the estimated albedo, the general problem of estimating the shape of an object with varying albedo map and unknown illumination source is reduced to one that can be handled by traditional SFS approaches. Experimental results are provided to show the effectiveness of the approach and its application to illumination-invariant matching and shape recovery. The estimated albedo maps are compared with the ground truth. The maps are used as illumination-invariant signatures for the task of face recognition across illumination variations. The recognition results obtained compare well with the current state-of-the-art approaches. Impressive shape recovery results are obtained using images downloaded from the Web with little control over imaging conditions. The recovered shapes are also used to synthesize novel views under novel illumination conditions. PMID- 19299863 TI - Transitions of the 3D medial axis under a one-parameter family of deformations. AB - The instabilities of the medial axis of a shape under deformations have long been recognized as a major obstacle to its use in recognition and other applications. These instabilities, or transitions, occur when the structure of the medial axis graph changes abruptly under deformations of shape. The recent classification of these transitions in 2D for the medial axis and for the shock graph was a key factor in the development of an object recognition system where the classified instabilities were utilized to represent deformation paths. The classification of generic transitions of the 3D medial axis could likewise potentially lead to a similar representation in 3D. In this paper, these transitions are classified by examining the order of contact of spheres with the surface, leading to an enumeration of possible transitions which are then examined on a case-by-case basis. Some cases are ruled out as never occurring in any family of deformations, while others are shown to be nongeneric in a one-parameter family of deformations. Finally, the remaining cases are shown to be viable by developing a specific example for each. Our work is inspired by that of Bogaevsky, who obtained the transitions as part of an investigation of viscosity solutions of Hamilton-Jacobi equations. Our contribution is to give a more down-to-earth approach, bringing this work to the attention of the computer vision community, and to provide explicit constructions for the various transitions using simple surfaces. We believe that the classification of these transitions is vital to the successful regularization of the medial axis in its use in real applications. PMID- 19299864 TI - Visual tracking by continuous density propagation in sequential bayesian filtering framework. AB - Particle filtering is frequently used for visual tracking problems since it provides a general framework for estimating and propagating probability density functions for nonlinear and non-Gaussian dynamic systems. However, this algorithm is based on a Monte Carlo approach and the cost of sampling and measurement is a problematic issue, especially for high-dimensional problems. We describe an alternative to the classical particle filter in which the underlying density function has an analytic representation for better approximation and effective propagation. The techniques of density interpolation and density approximation are introduced to represent the likelihood and the posterior densities with Gaussian mixtures, where all relevant parameters are automatically determined. The proposed analytic approach is shown to perform more efficiently in sampling in high-dimensional space. We apply the algorithm to real-time tracking problems and demonstrate its performance on real video sequences as well as synthetic examples. PMID- 19299865 TI - Estimating 3D positions and velocities of projectiles from monocular views. AB - In this paper, we consider the problem of localizing a projectile in 3D based on its apparent motion in a stationary monocular view. A thorough theoretical analysis is developed, from which we establish the minimum conditions for the existence of a unique solution. The theoretical results obtained have important implications for applications involving projectile motion. A robust, nonlinear optimization-based formulation is proposed, and the use of a local optimization method is justified by detailed examination of the local convexity structure of the cost function. The potential of this approach is validated by experimental results. PMID- 19299866 TI - Skeletal shape abstraction from examples. AB - Learning a class prototype from a set of exemplars is an important challenge facing researchers in object categorization. Although the problem is receiving growing interest, most approaches assume a one-to-one correspondence among local features, restricting their ability to learn true abstractions of a shape. In this paper, we present a new technique for learning an abstract shape prototype from a set of exemplars whose features are in many-to-many correspondence. Focusing on the domain of 2D shape, we represent a silhouette as a medial axis graph whose nodes correspond to "parts" defined by medial branches and whose edges connect adjacent parts. Given a pair of medial axis graphs, we establish a many-to-many correspondence between their nodes to find correspondences among articulating parts. Based on these correspondences, we recover the abstracted medial axis graph along with the positional and radial attributes associated with its nodes. We evaluate the abstracted prototypes in the context of a recognition task. PMID- 19299867 TI - Simultaneous localized feature selection and model detection for gaussian mixtures. AB - In this paper, we propose a novel approach of simultaneous localized feature selection and model detection for unsupervised learning. In our approach, local feature saliency, together with other parameters of Gaussian mixtures, are estimated by Bayesian variational learning. Experiments performed on both synthetic and real-world data sets demonstrate that our approach is superior over both global feature selection and subspace clustering methods. PMID- 19299868 TI - Significant progress in predicting the crystal structures of small organic molecules--a report on the fourth blind test. AB - We report on the organization and outcome of the fourth blind test of crystal structure prediction, an international collaborative project organized to evaluate the present state in computational methods of predicting the crystal structures of small organic molecules. There were 14 research groups which took part, using a variety of methods to generate and rank the most likely crystal structures for four target systems: three single-component crystal structures and a 1:1 cocrystal. Participants were challenged to predict the crystal structures of the four systems, given only their molecular diagrams, while the recently determined but as-yet unpublished crystal structures were withheld by an independent referee. Three predictions were allowed for each system. The results demonstrate a dramatic improvement in rates of success over previous blind tests; in total, there were 13 successful predictions and, for each of the four targets, at least two groups correctly predicted the observed crystal structure. The successes include one participating group who correctly predicted all four crystal structures as their first ranked choice, albeit at a considerable computational expense. The results reflect important improvements in modelling methods and suggest that, at least for the small and fairly rigid types of molecules included in this blind test, such calculations can be constructively applied to help understand crystallization and polymorphism of organic molecules. PMID- 19299869 TI - Ln3M(1-delta)TX7--quasi-isostructural compounds: stereochemistry and silver-ion motion in the Ln3Ag(1-delta)GeS7 (Ln = La-Nd, Sm, Gd-Er and Y; delta = 0.11-0.50) compounds. AB - The crystal structures of the Ln3Ag(1-delta)GeS7 (Ln = La-Nd, Sm, Gd-Er, Y; delta = 0.11-0.50, space group P6(3)) compounds were determined by means of X-ray single-crystal diffraction and the similarities among the crystal structures of all Ln3M(1-delta)TX7 (space group P6(3); Ln--lanthanide element, M--monovalent element; T--tetravalent element and X--S, Se) compounds deposited in the Inorganic Crystal Structure Database (ICSD) are discussed. Substitutions of each element in Ln3M(1-delta)TX7 result in a different structural effect. On the basis of the data deposited in the ICSD the large family of the Ln3M(1-delta)TX7 compounds was divided into three groups depending on the position of the monovalent element in the lattice. This position determines what kind of stereoisomer is present in the structure, either the ++ enantiomer or the +- diastereoisomer. Since the silver ions can occupy a different position and the energy barriers between positions are low the ions can move through the channel. It was shown that this movement is not a stochastic process but a correlated one. PMID- 19299870 TI - Symmetry rules and strain/order-parameter relationships for coupling between octahedral tilting and cooperative Jahn-Teller transitions in ABX3 perovskites. I. Theory. AB - Space groups, order-parameter and strain/order-parameter coupling relationships in ABX3 perovskite structures which combine cooperative Jahn-Teller distortions and octahedral tilting have been investigated from the perspective of group theory using the computer program ISOTROPY. Two common Jahn-Teller ordering schemes are associated with the irreducible representations M2+ and R3+ of the space group Pm3m. A third, less-common ordering scheme is associated with Gamma3+. These combine with tilting instabilities associated with M3+ and R4+ to generate a predicted suite of Jahn-Teller structure types that includes many of the known structures of manganites, vanadates, Cu and Cr halides. Order-parameter coupling and possible phase transitions are described using Landau free-energy expansions, and general expressions for the relationships between symmetry adapted spontaneous strains and particular order-parameter components are presented. These provide a general formal framework for determining structural evolution across multi-component order-parameter space and for characterizing the influence of tilting instabilities on Jahn-Teller instabilities or of Jahn-Teller ordering on octahedral tilting. PMID- 19299871 TI - Symmetry rules and strain/order-parameter relationships for coupling between octahedral tilting and cooperative Jahn-Teller transitions in ABX3 perovskites. II. Application. AB - The structural evolution of selected perovskites containing Jahn-Teller cations has been investigated in the light of a formal analysis of symmetry hierarchies for phase transitions driven by octahedral tilting and Jahn-Teller cooperative distortions. General expressions derived from the strain/order-parameter coupling relationships allowed by symmetry are combined with observed changes in lattice parameters to reveal details of order-parameter evolution and coupling. LuVO3, YbVO3, YVO3 and CeVO3 are representative of systems which develop Jahn-Teller ordering schemes associated with irreducible representations M2+ and R3+ of the space group Pm3m. Tilting of their octahedra is associated with M3+ and R4+. The Pnma (M3+ + R4+ tilting) <--> P2(1)/a (M3+ + R4+ tilting, R3+ Jahn-Teller order) transition below room temperature is close to second order in character. Shear strains which depend primarily on tilt angles show little variation, implying that there is only weak coupling between the tilting and Jahn-Teller order parameters. The subsequent P2(1)/a <--> Pnma (M3+ + R4+ tilting, M2+ Jahn-Teller order) is first order in character, and involves either a reduction in the R4+ tilt angle or a change in the strength of tilt/Jahn-Teller order-parameter coupling. In LaMnO3, the isosymmetric Pnma (M3+ + R4+ tilting) <--> Pnma (M3+ + R4+ tilting, M2+ Jahn-Teller order) transition can be described in terms of a classical first-order transition conforming to a 246 Landau expansion with negative fourth-order coefficients. Strain evolution in Ba-doped samples suggests that the transition becomes second order in character and reveals a new strain relaxation mechanism in LaMnO3 which might be understood in terms of local strain heterogeneities due to the disordering of distorted MnO6 octahedra. Transitions in PrAlO3 and La(0.5)Ba(0.5)CoO3 illustrate the transformation behaviour of systems in which the Jahn-Teller ordering scheme is associated with the irreducible representation Gamma3+. Overall, coupled tilting + Jahn-Teller phase transitions in perovskites conform to mean-field behaviour, consistent with the underlying role of strain in promoting long interaction lengths. PMID- 19299872 TI - X-ray powder diffraction and electron diffraction studies of the thortveitite related L phase, (Zn,Mn)2V2O7. AB - The phase designated gamma-Zn3(VO4)2 reported as a minor second phase in zinc oxide-based varistor materials doped with vanadium oxide and manganese oxide is shown to be the L phase, (Zn(1-x)Mn(x))2V2O7 (0.188 < x < 0.538), in the pseudo binary Mn2V2O7-Zn2V2O7 system. Analysis of X-ray powder diffraction patterns and electron diffraction patterns of this phase shows that the previously published a, c and beta values for this thortveitite-related phase are incorrect. Instead, Rietveld refinement of the X-ray powder pattern of the L phase shows that it has a monoclinic C lattice with Z = 6, with a = 10.3791 (1), b = 8.5557 (1), c = 9.3539 (1) A and beta = 98.467 (1) degrees. Although prior convergent-beam electron diffraction work of 'gamma-Zn3(VO4)2' confirmed the C Bravais lattice, the space group was found to be Cm rather than C2/m, the difference perhaps arising from the inability of the X-rays to detect small displacements of oxygen. Attempts to refine the structure in Cm did not produce improved R factors. The relationship between the crystal structure of the L phase and the high temperature C2/m beta'-Zn2V2O7 thortveitite-type solid solution is discussed. PMID- 19299873 TI - Polymorphism of CaTeO3 and solid solutions Ca(x)Sr(1-x)TeO3. AB - Single crystals and microcrystalline samples of the calcium tellurate(IV) phases alpha-, beta-, beta'- and gamma-CaTeO3 as well as of two solid solutions Ca(x)Sr(1-x)TeO3 (x = 0.55 and 0.77) have been synthesized and characterized by X ray diffraction and thermal analysis. A comparative description of the structures and the relations between the polymorphs is given. The main building units of the hitherto unknown structures are isolated [Te(IV)O3]2- units and [(Ca,Sr)O(x)] (x = 6-8) polyhedra. All structures exhibit channels in which the Te(IV) electron lone pairs protrude. The low-temperature phase alpha-CaTeO3 is stable up to 1168 K. It exhibits nearly cylindrical channels (diameter approximately 4 A) and differs structurally from the other phases, whereas the metastable high temperature phases are closely related to each other. They feature oval channels (shortest and longest diameter approximately 2 and 8 A). Gamma-CaTeO3 can be described as an order-disorder (OD) structure of two non-polar layers with layer groups p12(1)/m1 and p12(1)1. The gamma-CaTeO3 crystal under investigation consists of two polytypes with a maximum degree of order. The two phases Ca(x)Sr(1-x)TeO3 (x = 0.55 and 0.77) are isostructural to the MDO1 polytype of gamma-CaTeO3. Beta-CaTeO3 shows a distinct reversible phase transition at approximately 293 K. The low-temperature modification beta'-CaTeO3 as well as its high-temperature modification beta-CaTeO3 can be considered as threefold superstructures along [100] based on the MDO1 polytype of gamma-CaTeO3. PMID- 19299874 TI - Polymorphism in iodotris(tri-p-tolylphosphine)silver(I). AB - The reaction of silver(I) iodide with tri(p-tolyl)phosphine in MeCN solution in 1:3 molar ratio yields a polymorph of the complex of the formula [AgI{P(4 MeC6H4)3}3], with the Ag atom in a distorted tetrahedral environment. A polymorphic structure of this complex (a) is compared with previously published crystal structures (b), determined at different temperatures. The two polymorphs are compared using r.m.s. overlay calculations as well as half-normal probability plots. PMID- 19299875 TI - Electron diffraction, X-ray powder diffraction and pair-distribution-function analyses to determine the crystal structures of Pigment Yellow 213, C23H21N5O9. AB - The crystal structure of the nanocrystalline alpha phase of Pigment Yellow 213 (P.Y. 213) was solved by a combination of single-crystal electron diffraction and X-ray powder diffraction, despite the poor crystallinity of the material. The molecules form an efficient dense packing, which explains the observed insolubility and weather fastness of the pigment. The pair-distribution function (PDF) of the alpha phase is consistent with the determined crystal structure. The beta phase of P.Y. 213 shows even lower crystal quality, so extracting any structural information directly from the diffraction data is not possible. PDF analysis indicates the beta phase to have a columnar structure with a similar local structure as the alpha phase and a domain size in column direction of approximately 4 nm. PMID- 19299876 TI - Structures of six industrial benzimidazolone pigments from laboratory powder diffraction data. AB - The crystal structures of six industrially produced benzimidazolone pigments [Pigment Orange 36 (beta phase), Pigment Orange 62, Pigment Yellow 151, Pigment Yellow 154 (alpha phase), Pigment Yellow 181 (beta phase) and Pigment Yellow 194] were determined from laboratory X-ray powder diffraction data by means of real space methods using the programs DASH and MRIA, respectively. Subsequent Rietveld refinements were carried out with TOPAS. The crystal phases correspond to those produced industrially. Additionally, the crystal structures of the non-commercial compound 'BIRZIL' (a chloro derivative of Pigment Yellow 194) and of a dimethylsulfoxide solvate of Pigment Yellow 154 were determined by single-crystal structure analyses. All eight crystal structures are different; the six industrial pigments even exhibit five different hydrogen-bond topologies. Apparently, the good application properties of the benzimidazolone pigments are not the result of one specific hydrogen-bonding pattern, but are the result of a combination of efficient molecular packing and strong intermolecular hydrogen bonds. PMID- 19299877 TI - Structure determination of seven phases and solvates of Pigment Yellow 183 and Pigment Yellow 191 from X-ray powder and single-crystal data. AB - The crystal structures of two industrially produced laked yellow pigments, Pigment Yellow 183 [P.Y. 183, Ca(C16H10Cl2N4O7S2), alpha phase] and Pigment Yellow 191 [P.Y. 191, Ca(C17H13ClN4O7S2), alpha and beta phases], were determined from laboratory X-ray powder diffraction data. The coordinates of the molecular fragments of the crystal structures were found by means of real-space methods (simulated annealing) with the program DASH. The coordinates of the calcium ions and the water molecules were determined by combining real-space methods (DASH and MRIA) and repeated Rietveld refinements (TOPAS) of the partially finished crystal structures. TOPAS was also used for the final Rietveld refinements. The crystal structure of beta-P.Y. 183 was determined from single-crystal data. The alpha phases of the two pigments are isostructural, whereas the beta phases are not. All four phases exhibit a double-layer structure, built from nonpolar layers containing the C/N backbone and polar layers containing the calcium ions, sulfonate groups and water molecules. Furthermore, the crystal structures of an N,N-dimethylformamide solvate of P.Y. 183, and of P.Y. 191 solvates with N,N dimethylformamide and N,N-dimethylacetamide were determined by single-crystal X ray analysis. PMID- 19299878 TI - An aza-cyclophane stacked in racemic columnar assemblies: whole-molecule disorder in a two-dimensional solid solution. AB - The oxime derivative of [1.1.1]cyclophane cyclotriveratrylene (CTV) was ring expanded utilizing a Beckmann rearrangement to provide a ten-membered N-acetyl macrocyclic amide that crystallizes as a chloroform monosolvate in columnar assemblies manifesting an unusual disorder within the crystal. Columns made up of this structure consist of infinite columnar assemblies of alternating D and L enantiomers and therefore necessarily are made up of a racemate, yet the chiralities of individual molecules in adjacent columns are independent of one another, leading to the overall formation of a two-dimensional solid solution. The random arrangement of the columns within the structure leads to the emergence of a crystallographic mirror plane not reflected by the molecular symmetry, to a change of symmetry from Pna2(1) to Pnma and to whole-molecule disorder of the bowl-shaped molecules within the columns. PMID- 19299879 TI - Trans-cinnamic acid and coumarin-3-carboxylic acid: experimental charge-density studies to shed light on [2 + 2] cycloaddition reactions. AB - As part of an ongoing series of experimental charge-density investigations into the intra- and intermolecular interactions present in compounds which undergo solid-state [2 + 2] cycloaddition reactions, the charge-density analyses of trans cinnamic acid and coumarin-3-carboxylic acid are reported. Thus, high-resolution single-crystal X-ray diffraction data were recorded at 100 K for trans-cinnamic acid (sin theta/lambda(max) = 1.03 A(-1)) and coumarin-3-carboxylic acid (sin theta/lambda(max) = 1.19 A(-1)). In addition to the anticipated O-H...O hydrogen bonds weak C-H...O interactions were identified in both structures along with very weak intermolecular interactions between pairs of molecules that undergo solid-state [2 + 2] cycloaddition reactions upon irradiation. PMID- 19299880 TI - X-ray crystal structures of diacetates of 6-s-cis and 6-s-trans astaxanthin and of 7,8-didehydroastaxanthin and 7,8,7',8'-tetradehydroastaxanthin: comparison with free and protein-bound astaxanthins. AB - The crystal structures of the 6-s-cis [s-cis-(1)] and 6-s-trans [s-trans-(1)] conformers of the diacetates of astaxanthin (AXT) and those of (3S,3'S)-7,8 didehydroastaxanthin [(3S,3'S)-3,3'-dihydroxy-7,8-didehydro-beta,beta-carotene 4,4'-dione (2)] and (3S,3'S)-7,8,7',8'-tetradehydroastaxanthin [(3S,3'S)-3,3' dihydroxy-7,8,7',8'-tetradehydro-beta,beta-carotene-4, 4'-dione (3)] are reported. The conformations of these four molecules vary in particular with the angle of twist of the end rings out of the plane of the polyene chain; for s-cis (1), the end rings are twisted out of the plane of the polyene chain by an angle of -49.0 (5)degrees , and the conformation is therefore similar to that found for unesterified AXT as well as for the carotenoids, canthaxanthin and beta,beta carotene. For s-trans-(1), the end rings are coplanar with the polyene chain and its conformation is much more similar to that of the protein-bound AXT in the blue protein, crustacyanin, which is found in the shell of lobsters, although s trans-(1) shows much less bowing of the polyene chain. In (2) and (3) the end rings are also almost coplanar with the polyene chain with the end rings in (2) in the s-cis conformation, and in (3) in the s-trans conformation. Thus, an extensive ensemble of the possible beta end-ring conformations has been determined. These structures are compared with one another as well as unbound, unesterified AXT and protein-bound AXT. Also, the effect of the end-ring conformations on the colour and UV-vis spectra of the crystals was established. PMID- 19299881 TI - Mapping multiple dimensions of student learning: the ConstructMap program. AB - In the past, many assessments, especially standardized assessments, tended to be composed of items with specific right and wrong answers, such as those found in multiple choice, true-false and short response items. Performance-based questions that require students to construct answers rather than select correct responses introduce the complexities of multiple correct answers, dependence on teacher judgment for scoring, and requisite ancillary skills such as language fluency, which are technically difficult to handle, and may even introduce problems such as bias against certain groups of students. Recent developments in assessment design and psychometrics have improved the feasibility of assessing performance based tasks more efficiently and effectively, thereby providing a rich domain of information from which interpretations can be made about what students know and what they can do when they draw upon that knowledge. We developed the ConstructMap computer program specifically to assist teachers in interpreting and representing this type of performance data. The program accepts as input student scores on items associated with one or multiple performance variables, computes proficiencies using multidimensional item response methods, and produces graphical representations of students' estimated proficiency on each of the variables. PMID- 19299882 TI - Response dependence and the measurement of change. AB - Because of confounding effects that can mask change when persons respond to the same items on more than one occasion, the measurement of change is a challenge. The specific effect on change studied in this paper is that observed when responses of persons to items at time 2 are dependent statistically on their responses at time 1. In addition, because this response dependence may affect the change differently for different locations of items relative to persons at time 1, the initial targeting of persons to items was studied. For a specific change in means of persons, dichotomous data were simulated according to the Rasch model with varying degrees of dependence and varying initial targeting of persons to items. Data were analysed, also using the Rasch model, in two ways: firstly, by treating items used at time 1 and time 2 as distinct ones (rack analysis) and, secondly, by treating persons at time 1 and time 2 as distinct ones (stack analysis). With the rack analysis the change is revealed through the item parameters and with the stack analysis the change is revealed through the person parameters. With no response dependence the two analyses gave equivalent and correct measures of change. With increasing dependence change was increasingly masked or increasingly amplified, depending on the targeting of items to persons at time 1. Response dependence affected the measurement of change in both analyses, but not always in the same way. The paper serves as a warning against undetected dependence and also considers evidence that can be used in the analysis of real data sets for detecting the presence of dependence when measuring change. PMID- 19299883 TI - Using paired comparison matrices to estimate parameters of the partial credit Rasch measurement model for rater-mediated assessments. AB - The purpose of this paper is to describe a technique for estimating the parameters of a Rasch model that accommodates ordered categories and rater severity. The technique builds on the conditional pairwise algorithm described by Choppin (1968, 1985) and represents an extension of a conditional algorithm described by Garner and Engelhard (2000, 2002) in which parameters appear as the eigenvector of a matrix derived from paired comparisons. The algorithm is used successfully to recover parameters from a simulated data set. No one has previously described such an extension of the pairwise algorithm to a Rasch model that includes both ordered categories and rater effects. The paired comparisons technique has importance for several reasons: it relies on the separability of parameters that is true only for the Rasch measurement model; it works in the presence of missing data; it makes transparent the connectivity needed for parameter estimation; and it is very simple. The technique also shares the mathematical framework of a very popular technique in the social sciences called the Analytic Hierarchy Process (Saaty, 1996). PMID- 19299884 TI - Toward a domain theory in English as a second language. AB - This paper demonstrates how domain theory development is enhanced by using both theoretical data and empirical data. The study explored the domain of speaking English as a second language (ESL) comparing hypothetical data on speaking tasks provided by an experienced teacher and by a certified ACTFL oral proficiency interview rater with observed data from scores on a computer-delivered speaking exam. While the hypothetical data and observed data showed similar patterns in task difficulty in general, some tasks were identified as being much easier or harder than expected. These differences raise questions not only about test task design but also about the theoretical underpinnings of the domain. The results of the study suggest that this approach, where theory and data are examined together, will improve test design as well as benefit domain theory development. PMID- 19299885 TI - Comparison of single- and double-assessor scoring designs for the assessment of accomplished teaching. AB - This article is based on a more extensive research report (Engelhard, Myford and Cline, 2000) prepared for the National Board for Professional Teaching Standards (NBPTS) concerning the Early Childhood/Generalist and Middle Childhood/Generalist assessment systems. The report is available from the Educational Testing Service (ETS). An earlier version of the article was presented at the American Educational Research Association Conference in New Orleans in 2000. We would like to acknowledge the helpful advice of Mike Linacre regarding the use of the FACETS computer program and the assistance of Fred Cline in analyzing these data. The material contained in this article is based on work supported by the NBPTS. Any opinions, findings, conclusions, and recommendations expressed herein are those of the authors and do not necessarily reflect the views of the NBPTS, Emory University, ETS, or the University of Illinois at Chicago. PMID- 19299886 TI - A Rasch model prototype for assessing vocabulary learning resulting from different instructional methods: a preschool example. AB - This study explored the effects of using Rasch modeling to analyze data on vocabulary knowledge of preschoolers who participated in repeated read-aloud events and hands-on science activities with their classroom teachers. A Rasch prototype for literacy research was developed and applied to the preschool data. Thirty-one target words were selected for analysis from three children's informational picture books on light and color. After different instructional activities, each child received scores on individual target words measured with a total of six assessments, including free response vocabulary tests and expressive and receptive picture vocabulary tests. Rasch modeling was used to assess the learning difficulty of target words in different instructional settings. Suggestions are made for applying Rasch modeling to classroom studies of instructional interventions. PMID- 19299887 TI - An empirical study on the relationship between teacher's judgments and fit statistics of the partial credit model. AB - The main purpose of the study was to investigate empirically the relationship between classroom teacher's judgment and the item and person fit-statistics of the partial credit model. In this study, classroom teacher's judgments were made intuitively checking each item's consistency with the general response pattern and each student's need for additional treatment or advice. The item and person fit statistics of the partial credit model were estimated using the WINSTEPS program (Linacre, 2003). The subjects of this study were 321 sixth grade students in 9 classrooms within 3 elementary schools in Seoul, Korea. For this research, a performance assessment test for 6th grade mathematics was developed. It consisted of 20 polytomous response items and its total scores ranged between 0 and 50. In addition, the 9 classroom teachers made their judgments for each item of the test and for each student in their own classroom. They judged intuitively using 4 categories; (1) well fit, (2) fit, (3) misfit, and (4) badly misfit for each item as well as each student. Their judgments were scored from 1 to 4 for each item as well as each student. There are two significant findings in this study. First, there is a statistically significant relationship between the classroom teacher's judgment and item fit statistic for each item (The median correlation coefficient between the teacher's judgment and the item outfit ZSTD is 0.61). Second, there is a statistically significant relationship between the teacher's judgment and the person fit statistic for each student (The median correlation coefficient between the teacher's judgment and the person outfit ZSTD is 0.52). In conclusion, the item and person fit statistics of the partial credit model correspond with the teacher's judgments for each test item and each student. PMID- 19299888 TI - Tools for measuring academic growth. AB - Growth measurement and growth modeling have gained substantial interest in the last few years with the development of new statistical procedures and policy decisions such as the incorporation of growth into No Child Left Behind. The current study investigates the following four aspects of growth measurement: a) issues in the development of vertical scales to measure growth; b) design of instruments to measure academic growth; c) techniques for modeling individual student growth, and d) uses of growth information in a classroom. Measuring growth has always been a daunting task, but the development of measurement tools such as the Rasch model and computerized adaptive testing position us well to obtain high-quality data with which to measure and model the growth of an individual student across a course of study. This growth information, in norm referenced and standards-referenced form, should enhance educators' ability to enrich student learning. PMID- 19299889 TI - Effect of preceding inspiratory speed and end-inspiratory pause on forced expiratory manoeuvre in healthy subjects and chronic obstructive pulmonary disease patients. AB - BACKGROUND: Lower peak expiratory flow (PEF) and forced expiratory volume in 1 s (FEV(1)) have been consistently found after slow inspiration with end-inspiratory pause (EIP). OBJECTIVES: It was the aim of this study to establish the respective influence of the speed of preceding inspiration (SPI) and EIP on the parameters obtained from the following expiratory forced vital capacity (FVC) manoeuvre. METHODS: In 8 healthy subjects and 12 patients with chronic obstructive pulmonary disease (COPD), a number of inspirations with different SPI and EIP were performed. In the subsequent FVC manoeuvre, maximal expiratory flows, including PEF, and maximal expired volumes at different times, including FEV(1), were measured. For each FVC manoeuvre, peak expiratory time, expired volume at PEF (as % of FVC), flow limitation by the negative expiratory pressure technique and FVC were checked to be sure of achieving a similar expiratory effort and starting inflation lung volume. RESULTS: The highest values of PEF and FEV(1) were found in normal subjects and COPD patients after fastest SPI without EIP (p < 0.001). In normal subjects, no significant PEF and FEV(1) changes during FVC manoeuvre were observed with different SPI, in the absence of EIP. In contrast, inspirations with slower SPI (inspiratory time >2 s) without EIP were followed by lower PEF in COPD patients (p < 0.05). As compared with inspirations without EIP, those with a presence of EIP were invariably followed by lower PEF and FEV(1), both in normal subjects and in COPD patients (p < 0.05). CONCLUSIONS: The effect of SPI on subsequent PEF and FEV(1) is irrelevant in healthy subjects as well as in COPD patients, unless SPI is too slow (inspiratory time >2 s), while any EIP decreases these indices in all individuals. PMID- 19299890 TI - Persistent cough in a young smoker. Coccidioidomycosis infection, pulmonary nodular presentation. PMID- 19299891 TI - E1A has no effect on LPS-induced IL-6 secretion in rat alveolar epithelial cells. AB - BACKGROUND: The adenoviral protein E1A has been suggested to play a role in the pathophysiological development of chronic obstructive pulmonary disease (COPD) by inducing expression of inflammatory factors. It is well known that glucocorticoids are important inhibitors of inflammation. In the treatment of COPD corticosteroid therapy commonly has little or no anti-inflammatory effect. We hypothesized that the anti-inflammatory effect of glucocorticoids may be decreased or abolished by E1A expression, which aggravates the airway inflammation and results in the late stage of 'corticosteroid resistance' in COPD development. Corticosteroid therapy is a widely used method to treat respiratory inflammation. However, loss of anti-inflammatory effect is a common and serious complication during the therapy for COPD. OBJECTIVES: To test whether the E1A gene, which is 1 of the viral genes responsible for enhanced inflammatory responses, is involved in corticosteroid resistance during clinical therapy for COPD. METHODS: Rat alveolar epithelial cells were transfected with an E1A gene vector. Stably transfected cells were screened by PCR amplification of the E1A gene fragment. Transcription of the E1A gene was assayed by QT-RT-PCR. Protein expression was measured by immunohistochemistry and Western blot. E1A induced interleukin (IL)-6 secretion. mRNA levels were tested by ELISA and QT-RT-PCR. E1A positive cells and E1A-negative cells were treated with lipopolysaccharide (LPS), physiological concentrations of hydrocortisone and trichostatin A (TSA), an inhibitor of histone deacetylase (HDAC). The activity of HDAC was confirmed by colorimetric HDAC activity assay kit. RESULTS: In all 4 E1A-expressing transfectants, expression of the E1A 243R protein was much higher than that of the 289R protein. LPS significantly increased IL-6 secretion in both E1A-positive and E1A-negative cells and no significant differences were found. Treatment with hydrocortisone at a physiological concentration clearly decreased LPS-induced IL 6 secretion, and this down-regulation effect could be abolished in the presence of TSA. LPS significantly decreased HDAC activity, which was completely abolished by application of hydrocortisone. Hydrocortisone alone had no effect on HDAC activity in cells without LPS stimulation. There were no differences in HDAC activity or IL-6 secretion under these treatments between E1A-positive and E1A negative cells. CONCLUSIONS: Our results indicated that a physiological concentration of glucocorticoid suppresses LPS-mediated IL-6 secretion, through the participation of HDAC. Expression of adenoviral E1A had no effect on HDAC activity and hydrocortisones induced an anti-inflammatory effect. PMID- 19299893 TI - Calcium and phosphorus homeostasis. AB - Calcium and phosphorus homeostasis relies on a complex, tightly regulated system involving many ions and hormones. The regulation of calcium and phosphorus is controlled by the actions of these ions and hormones on the intestine, kidneys and bone. Disturbances in the serum level of calcium and/or phosphorus can lead to significant pathology, including kidney stones and bone disease. In addition to parathyroid hormone and vitamin D, recently identified factors such as fibroblast growth factors and klotho play an important role in maintaining mineral ion homeostasis. The identification of subfamily V transient receptor potential cation channels (TRPV channels), Na/P(i) cotransporters, the vitamin D receptor and the calcium-sensing receptor have further advanced our understanding of this complex physiology. In this review we discuss the current understanding of the relationships between the ions, hormones, and transporters that maintain calcium and phosphorus homeostasis. PMID- 19299892 TI - Interaction of aldosterone and extracellular volume in the pathogenesis of obesity-associated kidney disease: a narrative review. AB - Obesity and obesity-associated kidney injuries have played an important role in the rising prevalence of chronic kidney disease (CKD). The link between obesity and kidney disease begins with obesity's well-known associations with diabetes and hypertension, the two leading etiologies of CKD. However, a growing body of evidence suggests that elevated aldosterone levels and expanded extracellular volume are key components of obesity-induced renal disease via aldosterone's non epithelial effects on the kidney. Highlighting these blood pressure- and diabetes independent mechanisms of kidney injury in obesity allows an exploration of whether mineralocorticoid receptor blockade, coupled with weight loss and salt restriction, is an optimal treatment for overweight CKD patients. PMID- 19299894 TI - Fibronectin gene expression in aortic regurgitation: relative roles of mitogen activated protein kinases. AB - OBJECTIVES: In aortic regurgitation (AR), fibronectin (FN) expression is upregulated. This study sought to determine signal transduction pathways involved in upregulation of FN expression in AR. METHODS: Cardiac fibroblasts (CF) from rabbits with surgically induced AR and matched controls (NL) were cultured and assayed for FN expression and kinase activity with and without inhibitors of kinases JNK, p38 mitogen-activated protein kinase (MAPK) and extracellular response kinase (ERK). NL CF also were subjected to cyclic strain mimicking AR for 24 h in culture with and without inhibitors. RESULTS: AR CF exhibited 2.9 fold greater c-Jun phosphorylation (p < 0.01) and 1.5- to 2-fold greater ATF2 phosphorylation (p < 0.05-0.01) than NL. JNK and p38MAPK inhibition reduced c-Jun and ATF2 phosphorylation to NL; ERK inhibition had no effect. FN mRNA expression was similar in pattern to kinase activities. Cyclic strain in NL CF increased c Jun phosphorylation 2-fold versus unstrained controls (p < 0.005). This was suppressed by inhibition of JNK but not p38MAPK. CONCLUSION: FN expression in response to the acute mechanical strain resembling AR is upregulated primarily via JNK. However, in chronic AR both JNK and p38MAPK are involved. These signaling pathways represent potential therapeutic targets for normalizing extracellular matrix (ECM) composition and contractile force transmission, believed to be related to ECM composition/organization, in AR. PMID- 19299895 TI - Atrial Na,K-ATPase increase and potassium dysregulation accentuate the risk of postoperative atrial fibrillation. AB - BACKGROUND: Postoperative atrial fibrillation is a common complication to cardiac surgery. Na,K-ATPase is of major importance for the resting membrane potential and action potential. The purpose of the present study was to evaluate the importance of Na,K-ATPase concentrations in human atrial biopsies and plasma potassium concentrations for the development of atrial fibrillation. METHODS: Atrial myocardial biopsies were obtained from 67 patients undergoing open chest cardiac surgery. Na,K-ATPase was quantified using vanadate-facilitated 3H-ouabain binding. Plasma potassium concentration was measured with ion-selective electrode. RESULTS: In patients with preoperative sinus rhythm, 3H-ouabain binding site concentration was 16% higher in patients developing postoperative atrial fibrillation compared to patients maintaining sinus rhythm [302 +/- 15 pmol/g wet weight (n = 20) vs. 261 +/- 11 mmol/g wet weight (n = 33), p = 0.03]. Also with multivariable analysis, 3H-ouabain-binding site concentration was significantly associated with the development of atrial fibrillation. High increase in plasma potassium concentration during the perioperative period and surgery was associated with postoperative atrial fibrillation. CONCLUSIONS: The present study supports the increasing evidence of dysregulation of the potassium homeostasis as an important factor in the development of cardiac arrhythmias. High atrial Na,K-ATPase and sudden plasma potassium concentration increase may contribute to precipitate atrial fibrillation. PMID- 19299896 TI - Validation of the Danish Addenbrooke's Cognitive Examination as a screening test in a memory clinic. AB - BACKGROUND: Addenbrooke's Cognitive Examination (ACE) is a cognitive screening test developed to detect dementia. It has been validated in several countries. Validation studies have predominantly included patients with various degrees of dementia and healthy controls. OBJECTIVE: The aim of this study was to evaluate the Danish version of ACE as a screening test for early dementia in an outpatient memory clinic. Further, we wanted to investigate the ability of the ACE to discriminate patients with early Alzheimer's disease (AD) from patients with depression. METHOD: 78 patients with mild AD (MMSE >or=20), 30 non-demented patients diagnosed with depression (originally referred for evaluation of cognitive symptoms), and 63 healthy volunteers, all between 60 and 85 years of age, were included. All patients were given the ACE as a supplement to the standard diagnostic work-up. RESULTS: The cut-off points for optimal trade-off between sensitivity and specificity for ACE were 85/86 (sensitivity 0.99, specificity 0.94). When these cut-off points were applied to the group of depressive patients, the specificity dropped to 0.64, indicating a great overlap in individual test scores for demented and depressed patients. CONCLUSION: The optimal cut-off points for ACE found in this Danish study were close to what is reported in most other European studies. The great overlap in ACE scores for demented and depressed patients emphasize that test scores must be interpreted with great caution when used in diagnostic work-up. PMID- 19299897 TI - Structural and functional causes of hypernasality in velocardiofacial syndrome. A pilot study. AB - OBJECTIVE: Hypernasality in velocardiofacial syndrome (VCFS) is more severe, persistent, and difficult to manage compared to other populations with cleft palate or velopharyngeal (VP) dysfunction. This pilot study investigated why children with VCFS have more severe hypernasality. METHODS: Pressure-flow methodology indirectly measured VP orifice size and VP closure timing during speech in a group of 5 children with VCFS, 5 children with cleft palate, and 6 normal children. RESULTS: Children with VCFS demonstrated significant differences in VP closure timing and hypernasality. There were no significant group differences in VP orifice size. Duration of nasal airflow was the strongest predictor of judgments of hypernasality. CONCLUSION: This study provides preliminary evidence that VP closure timing may account for the more severe hypernasality in children with VCFS, compared to structural factors alone. PMID- 19299898 TI - An acoustical and perceptual study of vowels produced by alaryngeal speakers of Cantonese. AB - OBJECTIVE: The present study determines the acoustical and perceptual differences between alaryngeal and laryngeal speakers of Cantonese in the production of Cantonese vowels. SUBJECTS AND METHODS: Formant frequencies (F1 and F2) associated with eight Cantonese vowels [see text] produced by 40 alaryngeal (10 esophageal, 10 tracheoesophageal, 10 with pneumatic artificial larynx, 10 electrolaryngeal) and 10 laryngeal speakers were obtained. To assess how formants affect vowel perception, the vowels were identified by 20 naive listeners, based on which confusion matrices were established. RESULTS: Spectral analysis revealed that F1 and F2 values of Cantonese vowels produced by the alaryngeal speakers were significantly higher than those produced by laryngeal speakers, with the exception of F2 of [see text], /i/ and /y/ produced by alaryngeal speakers using pneumatic artificial larynx, and [see text] produced by electrolaryngeal speakers. CONCLUSION: The acoustic findings suggest a general shortening of the effective vocal tract length for resonance in alaryngeal speakers after laryngectomy. Percent correct identification of vowels from the perceptual experiment indicated similar error patterns in vowel identification between alaryngeal and laryngeal speaker groups. Among the different alaryngeal speaker groups, naive listeners tended to perceive the vowels produced by electrolaryngeal speakers more accurately. PMID- 19299899 TI - Effect of psychological stress on female vocal quality. A multiparameter approach. AB - Psychological stress can influence vocal quality. The purpose of the present study was to determine the effect of stress on female vocal quality. In this experimental study 54 female voices were studied in a stress-inducing and a relaxed condition. In the stress-inducing condition the subjects were asked to read a passage while instructions were used to evoke stress. In the relaxed condition the same reading task but without specific instructions was used. Identical subjective and objective assessment techniques were used to determine vocal quality in the stressed and the relaxed conditions. In stress-inducing and challenging conditions the female voice is more breathy, strained and of a lower objective vocal quality characterized by lower fundamental frequency, lower frequency, intensity and aerodynamic capacity. This study indicates that stress inducing and challenging conditions change vocal quality and vocal performance. Understanding of the effect of stress on voice production is relevant for clinical diagnostic situations, particularly in patients who present with a functional (nonorganic) voice disorder, as well as those patients who have some organic pathology but who also report exacerbation of symptoms during periods of increased life stress. PMID- 19299901 TI - Survival and re-operation rates after neurosurgical procedures in Scotland: implications for targeted surveillance of sub-clinical variant Creutzfeldt-Jakob disease. AB - BACKGROUND/AIM: To assess the feasibility of post-mortem surveillance for subclinical variant Creutzfeldt-Jakob disease (vCJD) at least 5 years after neurosurgical procedures. METHODS: Using Scottish record linkage, we estimated 5 year survival and re-operation rates after 4 neurosurgical procedures performed during 1993-2001 and identified as high or medium risk for transmitting vCJD: [B] drainage of extra- or subdural haematoma, [E] primary or revisional decompression operations and [H] creation of other ventricular shunts were classified as high risk; [C] operations on cerebral aneurysm (clipping) were classified as medium risk. RESULTS: Fatality rate at 1 year depended strongly on procedure, weakly or not at all on sex and era, and increased with age. Procedure rates differed by sex. The rate of subsequent neurosurgical operations was highest for procedure [H] (sole: 21%; multiple: 28%). CONCLUSION: Each year, the UK has a new cohort of some 5,000 5-year survivors after a high- or medium-risk neurosurgical procedure, whose subsequent annual mortality is at least 3%. Even if half the surviving 5 year survivors of neurosurgery since 1996 gave consent-in-life for vCJD informative testing at post-mortem, there would be too few relevant post-mortems in 2008-2010 (around 1,600) for 'nil detections' to exclude a 1 in 1,000 subclinical vCJD rate. Autopsy surveillance beyond 2010, or among 5-year survivors of non-neurosurgical at-risk operations, would be needed. PMID- 19299900 TI - MRI correlates of protein deposition and disease severity in postmortem frontotemporal lobar degeneration. AB - BACKGROUND: Frontotemporal lobar degeneration (FTLD) can be classified based on the presence of the microtubule-associated protein tau and the TAR DNA binding protein-43 (TDP-43). Future treatments will likely target these proteins, therefore it is important to identify biomarkers to help predict protein biochemistry. OBJECTIVE: To determine whether there is an MRI signature pattern of tau or TDP-43 using a large cohort of FTLD subjects and to investigate how patterns of atrophy change according to disease severity using a large autopsy confirmed cohort of FTLD subjects. METHODS: Patterns of gray matter loss were assessed using voxel-based morphometry in 37 tau-positive and 44 TDP-43-positive subjects compared to 35 age and gender-matched controls, and compared to each other. Comparisons were also repeated in behavioral variant frontotemporal dementia (bvFTD) subjects (n = 15 tau-positive and n = 30 TDP-43-positive). Patterns of atrophy were also assessed according to performance on the Clinical Dementia Rating (CDR) scale and Mini-Mental State Examination (MMSE). RESULTS: The tau-positive and TDP-43-positive groups showed patterns of frontotemporal gray matter loss compared to controls with no differences observed between the groups, for all subjects and for bvFTD subjects. Patterns of gray matter loss increased in a graded manner by CDR and MMSE with loss in the frontal lobes, insula and hippocampus in mild subjects, spreading to the temporal and parietal cortices and striatum in more advanced disease. CONCLUSION: There is no signature pattern of atrophy for tau or TDP-43; however, patterns of atrophy in FTLD progress with measures of clinical disease severity. PMID- 19299902 TI - Risk factors for posterior compared to anterior ischemic stroke: an observational study of the Registry of the Canadian Stroke Network. AB - BACKGROUND: Traditional vascular risk factors appear to exert varying magnitudes of risk for different major vascular events. For example, hypercholesterolemia is a much stronger risk factor for myocardial infarction than ischemic stroke. Limited evidence also suggests that vascular risk factors may exert differing magnitudes of risk for ischemic stroke within different cerebral arterial territories. We sought to determine the association between traditional vascular risk factors and the location of ischemic stroke (posterior versus anterior). METHODS: Consecutive patients with acute ischemic stroke who were admitted to 11 regional stroke centers within the Registry of the Canadian Stroke Network were included in the study sample. The Oxfordshire Community Stroke Project classification was used to distinguish posterior from anterior circulation ischemic stroke. Multivariable logistic regression was applied to determine the association between risk factors (age, gender, diabetes mellitus, hypercholesterolemia, hypertension, atrial fibrillation and smoking history) and posterior (compared to anterior) circulation ischemic stroke. RESULTS: In total, 8,489 patients with acute ischemic stroke were included. On multivariable analysis, diabetes mellitus (OR = 1.14; 95% CI = 1.02-1.27) was associated with an increased odds of posterior circulation ischemic stroke, whereas age (OR = 0.86; 95% CI = 0.83-0.90), female sex (OR = 0.84; 95% CI = 0.76-0.93), atrial fibrillation (OR = 0.83; 95% CI = 0.74-0.94) and pulmonary edema (OR = 0.74; 95% CI = 0.62-0.88) were related to a reduced odds of posterior compared with anterior circulation ischemic stroke. CONCLUSIONS: Some traditional vascular risk factors for ischemic stroke appear to exert different magnitudes of risk for posterior compared to anterior circulation ischemic stroke. PMID- 19299903 TI - The role of UFT in metastatic colorectal cancer. AB - 5-Fluorouracil (5-FU) has been the most widely used chemotherapeutic agent for metastatic colorectal cancer (mCRC) and 5-FU combination therapy improves efficacy compared with monotherapy. The oral fluoropyrimidine UFT (tegafur uracil) with leucovorin (LV) improves tolerability and has replaced 5-FU in many regimens. The efficacy and tolerability of UFT with LV in the first-line treatment of mCRC has been demonstrated in a number of phase II studies. In two phase III studies, UFT with LV has been shown to have comparable efficacy and improved tolerability versus intravenous bolus 5-FU, with very few cases of hand foot syndrome (HFS). Indirect comparisons of UFT and capecitabine suggest that they are comparable in terms of survival. In first-line treatment, UFT in combination with oxaliplatin (TEGAFOX) or irinotecan (TEGAFIRI) is effective and well tolerated, with similar efficacy and tolerability to the corresponding 5-FU- and capecitabine-based combinations, but with a lower incidence of HFS. Alternating cycles of TEGAFOX and TEGAFIRI are effective and well tolerated, and the combination of TEGAFIRI and the targeted monoclonal antibody cetuximab has shown promising activity, similar to that of FOLFIRI plus cetuximab. UFT can be considered a rational replacement for intravenous 5-FU in the first- and second line treatment of patients with mCRC. PMID- 19299904 TI - Phase I pharmacokinetic study of weekly intravenous and intraperitoneal paclitaxel combined with S-1 for advanced gastric cancer. AB - OBJECTIVES: A dose-escalation study of weekly intraperitoneal paclitaxel (PTX) combined with S-1 and intravenous PTX was performed to determine the maximum tolerated dose (MTD) and recommended dose (RD) in gastric cancer patients. PATIENTS AND METHODS: Nine gastric cancer patients with peritoneal dissemination and/or cancer cells on peritoneal cytology were enrolled. PTX was administered intravenously on days 1 and 8 at a fixed dose of 50 mg/m(2), and intraperitoneally with an initial dose of 20 mg/m(2), stepped up to 30 or 40 mg/m(2). S-1 was administered at a fixed dose of 80 mg/m(2)/day for 14 consecutive days, followed by 7 days of rest. A pharmacokinetic study of PTX was also performed. RESULTS: The MTD was determined to be 30 mg/m(2), as 2 of 3 patients developed dose-limiting toxicities, grade 3 febrile neutropenia and diarrhea. Therefore, the RD was determined to be 20 mg/m(2). The intraperitoneal and serum PTX concentration remained effective for over 72 and 48 h, respectively. CONCLUSIONS: Combined chemotherapy of S-1 plus weekly intravenous and intraperitoneal PTX was shown to be a safe regimen that should be further explored in clinical trials. PMID- 19299905 TI - Clinical significance of UDP-glucuronosyltransferase 1A1*6 for toxicities of combination chemotherapy with irinotecan and cisplatin in gynecologic cancers: a prospective multi-institutional study. AB - BACKGROUND: To investigate the effects of UDP-glucuronosyltransferase 1A1 (UGT1A1) *28, *6 and *27 in patients with gynecologic cancer who received chemotherapy with irinotecan and cisplatin. METHODS: Patients eligible for this study had cervical or ovarian cancer treated with chemotherapy; a course of the regimen consisted of 60 mg/m(2) of irinotecan on days 1, 8 and 15, and 60 mg/m(2) of cisplatin on day 1 every 4 weeks. UGT1A1 polymorphisms and toxicities were analyzed. RESULTS: From March 2007 to December 2007, 30 Japanese patients were enrolled; 24 ovarian carcinoma patients and 6 cervical cancer patients. The following genotypes of UGT1A1 were found: wild type in 17 patients (57%), *28 in 4 patients (13%), *6 in 8 patients (27%), *28*6 in 1 case (3%) and no case of *27 (0%). Grade 3/4 neutropenia, thrombocytopenia and diarrhea were significantly more frequent in *6 patients compared with wild-type patients. Also, in *6 patients irinotecan administration on days 8 or 15 was significantly more often omitted due to toxicities. In patients with *28 or *28*6, side effects were similar to those in patients with *6. CONCLUSION: In addition to UGT1A1*28, UGT1A1*6 might also be a key candidate to determine the dose of combination chemotherapy with irinotecan and cisplatin. PMID- 19299906 TI - Evaluation of posterior pole with echography and optical coherence tomography in patients with Behcet's disease. AB - BACKGROUND: In Behcet's disease posterior segment involvement is very frequent (about 80%). Macular edema is the most commonly observed complication and is related to reduction of visual acuity. The purpose of the study was to evaluate choroidal involvement during posterior vasculitis due to Behcet's disease. METHODS: Thirty eyes (15 patients) presenting with posterior vasculitis due to Behcet's disease were studied. All patients had been evaluated with optical coherence tomography (Stratus OCT, Zeiss) to evaluate the retinal thickness (RT) and with standardized 8-MHz A scan echography to determine the retinochoroidal thickness (RCT). Echographic evaluation with 10- and 20-MHz probes was performed in all cases. RESULTS: In 7 eyes (23.3%), both RT and RCT were normal. In 5 eyes (16.6%), they were both increased. In the other 18 eyes (60%), the RCT was increased while the RT was normal, thus indicating a thickening of the choroid alone. No significant correlation was found between choroidal thickness and the presence of vitreal abnormalities. Neither increases in RT nor those in RCT were significantly related to visual acuity. No correlation was found between RT or RCT and pharmacological treatment. CONCLUSION: Choroidal inflammation could often play a primary etiological role during posterior vasculitis in Behcet's disease. Echographic detection of choroidal thickening in patients with Behcet's disease not presenting with retinal alterations or visual symptoms could represent a useful way to detect an early inflammation, in order to start a proper therapy and to influence the final visual prognosis. PMID- 19299907 TI - Scanning laser polarimetry with variable corneal compensation to detect preperimetric glaucoma using logistic regression analysis. AB - BACKGROUND/AIMS: To define if a new function improves the capability of the scanning laser polarimetry with variable corneal compensation (GDx-VCC) nerve fiber analyzer in detecting preperimetric glaucoma using 12 retinal nerve fiber layer (RNFL) peripapillary sectors. METHODS: Data were obtained in a cross sectional, hospital-clinic-based study in which 478 eyes from 478 glaucoma suspects were enrolled. All of them underwent ophthalmological examination, static automated perimetry (Humphrey 24-2 Swedish interactive threshold algorithm), optic nerve stereoscopic photography, red-free digital RNFL photography and GDx-VCC examination. The overall intake included 391 normal eyes and 60 preperimetric glaucoma eyes with defects in the fiber layer photographs. RESULTS: The mean values of inferior average, temporal-superior-nasal-inferior temporal average, superior average and the number (nerve fiber indicator) were significantly lower in the eyes with RNFL defects than in the control group, when compared with the obtained linear discriminating function. The area under the curve values for receiver-operating characteristics analysis of the new parameter were 5% better than the best GDx-VCC standard values. CONCLUSIONS: Our results confirm that the new function has a better diagnostic reliability in preperimetric glaucoma and confirms the usefulness of morphological diagnosis between normal and early damaged RNFLs. PMID- 19299908 TI - Focus is key to becoming an expert in any field of research. An interview with Prof. Howard A. Reber, chief of gastrointestinal surgery, and director of the Ronald S. Hirshberg Pancreatic Cancer Research Laboratory, University of California Los Angeles (UCLA), Los Angeles, Calif., USA. Interview by Martin E. Fernandez-Zapico. AB - Dr. Howard Reber is a world-renowned pancreatologist in the area of basic pancreatic physiology and the management of pancreatic diseases. He was leader in the development of the current recommendation for the treatment of pancreatic cancer. In this interview for Pancreatology, Dr. Reber shares his life experiences as a scientist in pancreatic research. and IAP. PMID- 19299909 TI - Diabetes. AB - Pancreatologists have often divided research of the pancreas based upon the origin of the function or disease, namely the endocrine or exocrine pancreas. In fact, as a result, many of our meetings and conferences have followed separate paths. Interestingly, among patients with chronic pancreatitis and pancreatic cancer, both disorders of the exocrine pancreas, diabetes is common. However, the clinical features of the diabetes associated with these two differ. Peripheral insulin resistance and hyperinsulinemia are the predominant diabetic traits in pancreatic cancer, while reduced islet cell mass and impaired insulin secretion are observed more often in chronic pancreatitis. The causal relationship between diabetes and pancreatic cancer remains an intriguing but unanswered question. Since diabetes often precedes pancreatic cancer, it is regarded as a potential risk factor for malignancy. On the other hand, there remains the possibility that pancreatic cancer secretes diabetogenic factors. Regardless of how the science ultimately illuminates this issue, there is increasing interest in utilizing screening for diabetes to aid early detection of pancreatic tumor lesions. Therefore, in this issue of Pancreatology and the Web, we explore the topic of diabetes to keep us alert to this very important association, even if we study diseases of the exocrine pancreas. PMID- 19299910 TI - Primers on molecular pathways--the insulin pathway. AB - The insulin pathway is crucial for the regulation of intracellular and blood glucose levels and the prevention of diabetes. Regulating blood glucose levels demands coordinated interaction between several tissues, which is facilitated by the release of and response to the hormone, insulin. In response to an increase in circulating glucose levels, insulin is secreted by pancreatic beta cells to cause an increase in the uptake of glucose, fatty acids and amino acids into adipose tissue, muscle and the liver to subsequently promote the storage of these nutrients in the form of glycogen, lipids and protein, respectively, as well as suppress hepatic glucose release. Insulin sensitivity is measured by the relative capacity of insulin to promote a decrease in blood glucose. Failure to uptake and store nutrients results in diabetes. Type-1 diabetes is characterized as an autoimmune disease, resulting in destruction of the insulin-producing beta cells and therefore an inability to synthesize insulin. In contrast, in type-2 diabetes, the body develops resistance to the biological actions of insulin, presumably due to defects in the insulin signaling pathway. In this 'Primers on Molecular Pathways', we discuss this important signaling pathway, which is central to pancreas biology. PMID- 19299911 TI - Modulation of the caveolin-3 localization to caveolae and STAT3 to mitochondria by catecholamine-induced cardiac hypertrophy in H9c2 cardiomyoblasts. AB - We investigated the effect of phenylephrine (PE)- and isoproterenol (ISO)-induced cardiac hypertrophy on subcellular localization and expression of caveolin-3 and STAT3 in H9c2 cardiomyoblast cells. Caveolin-3 localization to plasma membrane was attenuated and localization of caveolin-3 to caveolae in the plasma membrane was 24.3% reduced by the catecholamine- induced hypertrophy. STAT3 and phospho STAT3 were up-regulated but verapamil and cyclosporin A synergistically decreased the STAT3 and phospho- STAT3 levels in PE- and ISO-induced hypertrophic cells. Both expression and activation of STAT3 were increased in the nucleus by the hypertrophy. Immunofluorescence analysis revealed that the catecholamine- induced hypertrophy promoted nuclear localization of pY705-STAT3. Of interest, phosphorylation of pS727- STAT3 in mitochondria was significantly reduced by catecholamine-induced hypertrophy. In addition, mitochondrial complexes II and III were greatly down- regulated in the hypertrophic cells. Our data suggest that the alterations in nuclear and mitochondrial activation of STAT3 and caveolae localization of caveolin-3 are related to the development of the catecholamine induced cardiac hypertrophy. PMID- 19299912 TI - Shank 2 expression coincides with neuronal differentiation in the developing retina. AB - The retinal activity for vision requires a precise synaptic connectivity. Shank proteins at postsynaptic sites of excitatory synapses play roles in signal transmission into the postsynaptic neuron. However, the correlation of Shank 2 expression with neuronal differentiation in the developing retina remains to be elucidated regardless of previous evidences of Shank 2 expression in retina. Herein, we demonstrated that with progression of development, Shank 2 is initially detected in the inner plexiform layer at P2, and then intensively detected in inner plexiform layer, outer plexiform layer, and ganglion cell layer at P14, which was closely colocalized to the neurofilament expression. Shank 2 was, however, not colocalized with glial fibrillary acidic protein. Shank 2 expression was increased in the differentiated retinoblastoma cells, which was mediated by ERK 1/2 activation. Moreover, Shank 2 expression was colocalized with neurofilament at the dendritic region of cells. In conclusion, our data suggests that Shank 2 is expressed in the neurons of the developing retina and could play a critical role in the neuronal differentiation of the developing retina. PMID- 19299913 TI - Involvement of GADD153 and cardiac ankyrin repeat protein in cardiac ischemia reperfusion injury. AB - Oxidative stress is critical for causing cardiac injuries during ischemia reperfusion (IR), yet the molecular mechanism for this remains unclear. In the present study, we observe that hypoxia and reoxygenation, a component of ischemia, effectively induces apoptosis in the cardiac myocytes from neonatal rats and it concomitantly leads to induction of GADD153, an apoptosis-related gene. Furthermore, IR injury of rat heart showed a GADD153 overexpression in the ischemic area where the TUNEL reaction was positive. A downregulation of cardiac ankyrin repeat protein (CARP) was also observed in this ischemic area. Promoter deletion and reporter analysis revealed that hypoxia transcriptionally activates a GADD153 promoter through the AP-1 element in neonatal cardiomyocytes. Ectopic overexpression of GADD153 resulted in the downregulation of CARP expression. Accordingly, the induction of GADD153 mRNA were followed by the CARP down regulation in an in vivo rat coronary ischemia/reperfusion injury model. These results suggest that GADD153 over-expression and the resulting downregulation of CARP may have causative roles in apoptotic cell death during cardiac IR injury. PMID- 19299914 TI - Higher mitochondrial DNA copy number is associated with lower prevalence of microalbuminuria. AB - It has been suggested that mitochondrial dysfunction contributes to the initiation and development of atherosclerosis and cardiovascular disease. We examined the association between mitochondrial DNA (mtDNA) copy number and microalbuminuria in a cross-sectional community-based study. We measured peripheral blood mtDNA copy number in 694 adults without chronic kidney disease by a real-time PCR method. The overall prevalence of microalbuminuria (defined as an albumin creatinine ratio of 30 to 299 mg/g) was 4.5%. The prevalence of microalbuminuria decreased progressively from the lower to the upper quartiles of mtDNA copy number (6.9%, 5.7%, 2.9%, and 2.3% in quartiles 1, 2, 3, and 4, respectively, P=0.017 for trend). Multiple logistic regression models showed that the quartile of mtDNA copy number was independently associated with the prevalence of microalbuminuria (P=0.01 for trend). Compared with the lowest quartile, the highest quartile had an odds ratio of 0.22 for microalbuminuria (95% confidence interval, 0.05 to 0.87; P=0.03). Higher mtDNA copy number was associated with the lower prevalence of microalbuminuria in a community-based population. PMID- 19299915 TI - Regulation of expression of matrix metalloproteinase-9 by JNK in Raw 264.7 cells: presence of inhibitory factor(s) suppressing MMP-9 induction in serum and conditioned media. AB - Matrix metalloproteinase-9 (MMP-9) secreted from macrophages plays an important role in tissue destruction and inflammation through degradation of matrix proteins and proteolytic activation of cytokines/chemokines. Whereas the MEK-ERK and PI3K- Akt pathways up-regulate MMP-9 expression, regulation of MMP-9 by JNK remains controversial. Presently, we aimed to determine the role of JNK in MMP-9 regulation in Raw 264.7 cells. Inhibition of JNK by the JNK inhibitor SP600125 induced MMP-9 in the absence of serum and suppressed the expression of TNF-alpha, IL-6 and cyclooxygenase-2 in LPS-treated Raw 264.7 cells. In a knockdown experiment with small interfering RNA, suppression of JNK1 induced MMP-9 expression. Interestingly, mouse serum suppressed SP600125- mediated MMP-9 induction, similar to IFN-gamma. However, the inhibitory activity of mouse serum was not affected by pyridone 6, which inhibits Janus kinase downstream to IFN gamma. In addition to mouse serum, conditioned media of Raw 264.7 cells contained the inhibitory factor(s) larger than 10 kDa, which suppressed SP600125- or LPS induced MMP-9 expression. Taken together, these data suggest that JNK1 suppresses MMP-9 expression in the absence of serum. In addition, the inhibitory factor(s) present in serum or secreted from macrophages may negatively control MMP-9 expression. PMID- 19299916 TI - Calcium overload is essential for the acceleration of staurosporine-induced cell death following neuronal differentiation in PC12 cells. AB - Differentiation of neuronal cells has been shown to accelerate stress-induced cell death, but the underlying mechanisms are not completely understood. Here, we find that early and sustained increase in cytosolic ([Ca2(+)]c) and mitochondrial Ca2(+) levels ([Ca2(+)]m) is essential for the increased sensitivity to staurosporine- induced cell death following neuronal differentiation in PC12 cells. Consistently, pretreatment of differentiated PC12 cells with the intracellular Ca2(+)-chelator EGTA-AM diminished staurosporine-induced PARP cleavage and cell death. Furthermore, Ca2(+) overload and enhanced vulnerability to staurosporine in differentiated cells were prevented by Bcl-XL overexpression. Our data reveal a new regulatory role for differentiation-dependent alteration of Ca2(+) signaling in cell death in response to staurosporine. PMID- 19299917 TI - Simvastatin inhibits induction of matrix metalloproteinase-9 in rat alveolar macrophages exposed to cigarette smoke extract. AB - Matrix metalloproteinase-9 (MMP-9) may play an important role in emphysematous change in chronic obstructive pulmonary disease (COPD), one of the leading causes of mortality and morbidity worldwide. We previously reported that simvastatin, an inhibitor of HMG-CoA reductase, attenuates emphysematous change and MMP-9 induction in the lungs of rats exposed to cigarette smoke. However, it remained uncertain how cigarette smoke induced MMP-9 and how simvastatin inhibited cigarette smoke-induced MMP-9 expression in alveolar macrophages (AMs), a major source of MMP-9 in the lungs of COPD patients. Presently, we examined the related signaling for MMP-9 induction and the inhibitory mechanism of simvastatin on MMP 9 induction in AMs exposed to cigarette smoke extract (CSE). In isolated rat AMs, CSE induced MMP-9 expression and phosphorylation of ERK and Akt. A chemical inhibitor of MEK1/2 or PI3K reduced phosphorylation of ERK or Akt, respectively, and also inhibited CSE-mediated MMP-9 induction. Simvastatin reduced CSE-mediated MMP-9 induction, and simvastatin-mediated inhibition was reversed by farnesyl pyrophosphate (FPP) or geranylgeranyl pyrophosphate (GGPP). Similar to simvastatin, inhibition of FPP transferase or GGPP transferase suppressed CSE mediated MMP-9 induction. Simvastatin attenuated CSE-mediated activation of RAS and phosphorylation of ERK, Akt, p65, IkappaB, and nuclear AP-1 or NF-kappaB activity. Taken together, these results suggest that simvastatin may inhibit CSE mediated MMP-9 induction, primarily by blocking prenylation of RAS in the signaling pathways, in which Raf-MEK-ERK, PI3K/Akt, AP-1, and IkappaB-NF-kappaB are involved. PMID- 19299918 TI - Nickel induces secretion of IFN-gamma by splenic natural killer cells. AB - Although nickel hypersensitivity is known as a delayed-type hypersensitivity mediated by nickel-specific T cells, it is greatly influenced by other immune cells. Here we show that splenic natural killer cells (NK cells) directly or indirectly respond to nickel by secretion of IFN-gamma. Using enzyme-linked immunosorbent spot (ELISPOT) assays, we found that nickel-reactive cells readily secreted IFN-gamma when splenocytes were cultured in the presence of varying concentrations of nickel sulfate (NiSO(4)) for 24 h. However, nickel-reactive IL 2- or IL- 4-secreting cells were infrequent during the 24-h culture with NiSO(4). Immune responses to nickel were innate, not adaptive, in nature since the frequency of nickel-reactive IFN-g-secreting cells did not increase upon previous exposure to NiSO(4) and recombination activating gene (RAG)-1-deficient mice contained nickel-reactive IFN-gamma-secreting cells. The involvement of NK cells in the innate response to NiSO(4) was confirmed since we could observe a significant reduction of the frequency of nickel-reactive cells in NK cell depleted mice. Furthermore, the number of IFN-gamma secreting cells was significantly reduced in the ELISPOT assays when NKG2D was blocked by anti-NKG2D antibody. These results suggest that there is an early and rapid innate immune response to nickel, which is mediated by NK cells and the NKG2D receptor. The significance of the innate response to nickel is that it may contribute to development of the late T cell-mediated delayed type hypersensitivity against nickel. PMID- 19299969 TI - Semiprivate. AB - Dying patients should be given the choice, whenever possible, of spending their last hours in a private room. PMID- 19299990 TI - Medevac crashes on the rise. PMID- 19299991 TI - Nursing in Australia's remote territories. PMID- 19299992 TI - High-use supplies at the bedside. AB - This is the sixth in a series of articles from Massachusetts General Hospital in Boston describing one general medical unit's experiences with Transforming Care at the Bedside (TCAB). An initiative begun by the Robert Wood Johnson Foundation (RWJF) and the Institute for Healthcare Improvement, TCAB was developed as a way to improve care on medical-surgical units, patients' and family members' experience of care, and teamwork among care team members and to increase nurse satisfaction and retention. The TCAB philosophy engages all care leaders, but empowers bedside nurses to generate ideas and solutions for change. Mass General is one of 68 hospitals participating in a two-year TCAB initiative led by the American Organization of Nurse Executives and funded with a grant from the RWJF. For more information on TCAB, go to www.rwjf.org/pr/product.jsp?id=31512. PMID- 19299999 TI - Primary open-angle glaucoma. AB - Primary open-angle glaucoma (POAG), by far the most common form of glaucoma, currently afflicts more than 2 million Americans, more than half of whom probably don't know they have it. Asymptomatic in the early stages, it gradually and progressively reduces the visual field and leads to blindness if untreated. Elevated intraocular pressure is believed to play a role, yet some people with normal pressure develop POAG. There is no single diagnostic test; screening and diagnosis involve periodic comprehensive eye evaluations. Treatment is aimed at delaying onset, slowing progression, and preserving vision. PMID- 19300002 TI - Pheochromocytoma. PMID- 19300003 TI - Expressed breast milk: safety in the hospital. AB - The Pennsylvania Patient Safety Reporting System (PA-PSRS, pronounced PAY-sirs) is a confidential, statewide reporting system on the Internet to which all Pennsylvania hospitals, outpatient-surgery facilities, and birthing centers, as well as some abortion facilities, were required to file information on medical errors beginning in June 2004.Safety Monitor, this column in AJN from PA-PSRS, informs nurses on issues that can affect patient safety and presents strategies they can integrate easily into practice.For more information on PA-PSRS, visit the Web site of Pennsylvania's Patient Safety Authority, at www.psa.state.pa.us. For the original articles discussed in this column or for other articles on patient safety, click on "Advisories and Related Resources" in the left-hand navigation menu.This is a periodic column from the Pennsylvania Patient Safety Reporting System. PMID- 19300004 TI - Reducing health care's ecological footprint. PMID- 19300009 TI - How to try this: Assessment of transient urinary incontinence in older adults. AB - Urinary incontinence in older adults is associated with an increased risk of institutionalization, as well as with urinary tract infections and depression. Transient urinary incontinence arises suddenly, lasts less than six months, and results from reversible causes. Many caregivers erroneously consider urinary incontinence to be inevitable in older adults, especially in hospitalized patients. Failure to identify and respond to transient urinary incontinence may lead to established incontinence and to other poor outcomes after hospital discharge. A bladder diary and a mnemonic are two methods nurses can use to assess for transient urinary incontinence and its treatable underlying causes. For a free online video showing nurses using these methods, go to http://links.lww.com/A311. PMID- 19300013 TI - The Health Care Innovations Exchange. PMID- 19300014 TI - Health care reform in Massachusetts: is it working? PMID- 19300016 TI - Nurses and the pharmaceutical industry: part 2. PMID- 19300017 TI - Blessed messy work. PMID- 19300035 TI - Warfarin necrosis. PMID- 19300036 TI - Rehabilitation after scapulectomy. AB - An 84-yr-old man presented with right posterior shoulder pain and growing mass. Shoulder range of motion was limited in forward elevation and abduction; result of the neurologic testing was normal. Radiologic workup showed an irregular mass resembling Paget's disease or sarcoma. On surgical inspection, a 16-cm hemorrhagic mass, occupying the entire scapula, was identified. Patient underwent tumor resection and scapulectomy. Postoperatively, he started a rehabilitation program. With attempted ambulation, patient exhibited strong tendencies to lean rightward, walked in circles, and risking fall. The gait abnormality could not be corrected and acute inpatient rehabilitation admission was recommended. A unique rehabilitation program was developed to correct gait deviation, improve balance, and compensate for deficits. After a comprehensive inpatient rehabilitation, the patient was successfully discharged home. We present a review of scapulectomy, postoperative care, and a recommended rehabilitation program. PMID- 19300037 TI - Population pharmacokinetics of the selective serotonin 5-HT1A receptor partial agonist piclozotan. AB - BACKGROUND/AIMS: Serotonin (5-HT) and its receptors are known to play important roles in various physiological and pathophysiological processes. The 5-HT1A receptor subtype is thought to be involved in psychiatric disorders, immunomodulation, and in cerebral ischemic conditions. Piclozotan, a selective and potent partial agonist of 5-HT1A, has been shown to be neuroprotective against ischemic neuronal damage in animal models. Its pharmacokinetics (PK), tolerability, and safety have been evaluated in patients with acute ischemic stroke. The aim of the study was to describe piclozotan PK, using population PK modeling. METHODS: A total of 1308 plasma piclozotan concentration measurements from 84 healthy subjects and 412 plasma piclozotan measurements from 74 stroke patients were included in the analysis. Covariates considered during the model building process included disease status, age, weight, sex, smoking status, and alcohol consumption. Data were analyzed using nonlinear mixed-effects modeling with the NONMEM software system. The final model was qualified via predictive check and nonparametric bootstrap procedures. RESULTS: Piclozotan PK was well described using a 3-compartment model with first-order elimination. Parameter estimates (intersubject variability) were V1, central volume: 64.0 L (66.5%) and CL, systemic clearance: 18.0 L/h (31.4%). Peripheral volumes (V2, V3) were related to total body weight, whereas CL was related to ideal body weight. Clearance decreased with advancing age, yielding a decrease of 35%-65% in patients aged 70-90. There was no discernable difference in PK between healthy subjects and stroke patients. CONCLUSIONS: Piclozotan disposition was well described by the population model, and the intersubject variability around the estimated parameters was moderate in magnitude (<60%). The population PK analysis of piclozotan allows for characterization of piclozotan exposure in individual subjects based on their age and body weight. The availability of a population PK model will facilitate dose optimization and further clinical development of piclozotan. PMID- 19300038 TI - Effect of enhanced external counterpulsation on clinical symptoms, quality of life, 6-minute walking distance, and echocardiographic measurements of left ventricular systolic and diastolic function after 35 days of treatment and at 1 year follow up in 47 patients with chronic refractory angina pectoris. AB - In a prospective study, enhanced external counterpulsation (EECP) was performed for 1 hour each day for 35 days in 47 patients, mean age 61 +/- 8 years, with prior coronary revascularization who had chronic refractory angina pectoris despite antianginal drugs and who were not candidates for further coronary revascularization. Compared with baseline values, EECP significantly improved anginal symptoms, dyspnea on exertion, and quality of life after 35 days of treatment (P < 0.001) and at 1-year follow up (P < 0.001). Compared with the baseline value of 653 +/- 249 feet, EECP significantly improved the 6-minute walking distance to 1025 +/- 234 feet after 35 days of treatment (P < 0.001) and to 1040 +/- 221 feet at 1-year follow up (P < 0.001). However, EECP did not significantly affect left ventricular ejection fraction, left ventricular end diastolic and end-systolic dimensions, left ventricular end-diastolic and end systolic volumes, E/A ratio, isovolumic relaxation time, and deceleration time measured by two2-dimensional and Doppler echocardiography. PMID- 19300039 TI - Improvement in sensory pain rating after palliative systemic radionuclide therapy in patients with advanced prostate cancer. AB - This study assessed whether baseline and short-term patient-reported quality of life (QOL) differs in patients with symptomatic metastatic prostate cancer undergoing palliative management using opioids, nonsteroidal anti-inflammatory agents (NSAIDs), (89)strontium chloride ((89)Sr), and samarium-lexidronam ((153)Sm). Males were grouped according to primary palliative intervention: opioids (n = 40), NSAIDs (n = 40), (89)Sr chloride (n = 25), and (153)Sm (n = 25). The short form of the self-administered McGill Pain Questionnaire was used to measure QOL at baseline, 4 and 8 weeks after initiation of treatment. Clinical data were collected from patients' medical records. Statistical analyses were conducted using descriptive methods and the Student t test. A significant increase in the sensory pain rating was observed in the patients treated by NSAIDs ([upward arrow]21%) and (89)Sr ([upward arrow]46%), whereas those treated by opioids ([downward arrow]27%) and (153)Sm ([downward arrow]27%) demonstrated a significant (P < 0.05) decrease in this subscore. There was a longitudinal decrease in QOL over time in patients treated by NSAIDs and (89)Sr as measured by the total pain rating score, whereas those treated with the other agents experienced improved QOL. This study demonstrates improvement in QOL achieved using (153)Sm, which is comparable to that achieved with the use of opioids during this observation interval. PMID- 19300040 TI - Population pharmacokinetics of ketorolac in neonates and young infants. AB - Although ketorolac is commonly used as an analgesic in the pediatric population, there is no information on the pharmacokinetics of ketorolac available for children less than 6 months of age. The objective of this analysis was to construct a population pharmacokinetic model to describe ketorolac disposition in young children. Three neonates and 9 infants, median (range) age 0.4-32 weeks, were administered with 0.5 mg/kg of ketorolac. The data were best described by a 2-compartment model, with an allometric expression to describe body weight effects on clearance. Estimated parameters were clearance [2.8 mL x min(-1) x (kg(0.75))(-1)], intercompartmental clearance (11.5 mL/min), volume of distribution of the central compartment (535 mL), and volume of distribution of the peripheral compartment (322 mL). The clearance values in these neonates and younger group of infants are greater than that reported for older children and adults. PMID- 19300041 TI - Practical management of complex regional pain syndrome. AB - Complex regional pain syndrome (CRPS) describes a diversity of painful conditions following trauma, coupled with abnormal regulation of blood flow and sweating, trophic changes, and edema of skin. The excruciating pain and diverse autonomic dysfunctions in CRPS are disproportionate to any inciting and recovering event. CRPS type I is formerly identified as "reflex sympathetic dystrophy." CRPS type II is the new term for "causalgia" that always coexists with documented nerve injury. The present diagnostic criteria of CRPS I and II depend solely on meticulous history and physical examination without any confirmation by specific test procedure (or gold standard). There are only few clinical studies with large scale randomized trials of pharmacologic agents on the treatment of CRPS. Bisphosphonates have been studied in multiple controlled trials, based on theoretical benefit of bone resorption, to offer pain relief and functional improvement in patients with CRPS. Many current rationales in treatment of CRPS (such as topical agents, antiepileptic drugs, tricyclic antidepressants, and opioids) are mainly dependent on efficacy originate in other common conditions of neuropathic pain. There are additional innovative therapies on CRPS that are still in infancy. No wonder all the treatment of individual CRPS case nowadays is pragmatic at best. Although the interventional therapies in CRPS (such as nerve blockade, sympathetic block, spinal cord and peripheral nerve stimulation, implantable spinal medication pumps, and chemical and surgical sympathectomy) may offer more rapid response, yet it is still controversial with unpredictable outcome. Nevertheless, we need to start pain management immediately with the ambition to restore function in every probable case of CRPS. An interdisciplinary setting with comprehensive approach (pharmacologic, interventional, and psychological in conjunction with rehabilitation pathway) has been proposed as protocol in the practical management of CRPS. It is crucial to have a high sensitivity value combined with a fair specificity in revising diagnostic criteria of CRPS. The validation and consensus for new rationalized diagnostic criteria of CRPS could facilitate further research to enhance clinical outcome including quality of life. These endeavors to minimize suffering from CRPS would certainly be appreciated by many patients and their loved ones. PMID- 19300042 TI - Targeting the inflammatory response in secondary stroke prevention: a role for combining aspirin and extended-release dipyridamole. AB - The recognition that stroke and other ischemic events are manifestations of chronic progressive inflammation has had a great impact on the development of prevention strategies. The most recent American Heart Association guidelines recommend combination aspirin and extended-release dipyridamole over aspirin alone for patients with prior ischemic stroke or transient ischemic attack. Although aspirin and extended-release dipyridamole have long been recognized for their antiplatelet activities, there is now evidence that these drugs also have complementary antiinflammatory properties that contribute to improved outcomes when used to prevent secondary stroke. In the Second European Stroke Prevention Study (ESPS-2), the addition of extended-release dipyridamole to low-dose aspirin significantly reduced the risk of recurrent ischemic stroke without significantly increasing bleeding. Also, in the recent European/Australasian Stroke Prevention in Reversible Ischaemia Trial (ESPRIT), a combination of aspirin and extended release dipyridamole was superior to aspirin alone for reducing the occurrence of the primary combined end point of vascular death, nonfatal stroke, nonfatal myocardial infarction, and major bleeding complications. The added benefit without worsening bleeding may be attributable, in part, to the antiinflammatory actions of this combination therapy. PMID- 19300043 TI - Natriuretic peptide testing for heart failure therapy guidance in the inpatient and outpatient setting. AB - Acutely destabilized heart failure is one of the most common diagnoses in the modern health care system. It has high hospital readmission rates and significant short-, medium-, and long-term mortality, likely due to misdiagnosis or failure to assess adequate treatment before discharge. Cardiac biomarkers such as B-type natriuretic peptide and its amino terminal cleavage equivalent N-terminal fragment have rapidly become one of the key tools in the diagnosis and guidance of heart failure therapy. In this article, we shall review the data on the current use of the natriuretic peptides for the diagnosis, prognosis, and management of heart failure in both the outpatient and inpatient settings. PMID- 19300044 TI - Current treatment for cutaneous leishmaniasis: a review. AB - Cutaneous leishmaniasis is the most common form of leishmaniasis. It is a skin infection caused by a single-celled parasite that is transmitted by sand fly bites. There are about 20 species of Leishmania that may cause cutaneous leishmaniasis. Some Leishmania species are closely linked to humans and are therefore found in cities (Leishmania tropica), whereas some are more traditionally associated with animal species and are therefore considered zoonoses (Leishmania major). The evidence for optimal treatment of cutaneous leishmaniasis is patchy. Although the cutaneous form of the disease is often self limiting, it does result in significant scarring and can spread to more invasive, mucocutaneous disease. Therefore, treatment may be considered to prevent these complications. Drugs for systemic and topical treatment are presented and discussed with regard to their application, use, and adverse effects. PMID- 19300045 TI - Central sleep apnea in a geriatric patient treated with aripiprazole. AB - We report a case of central sleep apnea in a geriatric patient that was associated with treatment with aripiprazole for an episode of major depressive disorder with psychotic features. The patient was a 72-year-old man who was started on aripiprazole and developed central sleep apnea that improved significantly when the medication was stopped. A rechallenge with aripiprazole led to a worsening of the central sleep apnea, which again improved off the aripiprazole. We postulate that the central sleep apnea was due to aripiprazole. There have been numerous case reports in the literature of obstructive sleep apnea associated with atypical antipsychotics. To our knowledge, this is the first published case of central sleep apnea. We caution clinicians to be aware that there is potential risk of atypical antipsychotics like aripiprazole inducing or exacerbating central sleep apnea. PMID- 19300046 TI - Enhanced platelet responsiveness due to chilling and its relation to CD40 ligand level and platelet-leukocyte aggregate formation. AB - To investigate platelet responsiveness during cold storage of whole blood, we examined platelet aggregation, expression of CD40 ligand (CD40L) on platelets, the plasma levels of soluble form of CD40L (sCD40L) as well as platelet-leukocyte aggregates. Flow cytometry analysis was performed to investigate platelet leukocyte aggregate formation using antibodies against CD42b and CD45 and platelet activation using antibodies against P-selectin and PAC-1. Blood samples were collected from healthy volunteers, patients with cardiovascular diseases, or both. In the healthy volunteers' blood samples stored at 4 degrees C for 6 h, platelet aggregation in response to 1 micromol/l ADP was enhanced, and released levels of soluble form of P-selectin and thromboxane B2 in response to 1 micromol/l ADP markedly increased. In the samples stored at 4 degrees C for 6 h but not stimulated by any agonists, CD40L expression on the platelets was increased, and plasma levels of sCD40L were also elevated. Under the same condition, the increase in simultaneous expression of CD45 and CD42b was observed. In patients with cardiovascular diseases, the platelet aggregability, coexpression of P-selectin and PAC-1, expression of CD40L on platelets and both CD45-bound and CD42b-bound subsets were all comparable to those of healthy volunteers' samples stored at 4 degrees C for 6 h. Plasma levels of sCD40L in patients were higher than those in healthy volunteers' control. Taken together, storage of whole blood at 4 degrees C for 6 h caused platelet activation comparable to that of patients with cardiovascular diseases, and enhanced platelet activity in such patients may be involved in increased risk for thromboembolic events. PMID- 19300047 TI - Thromboangiitis obliterans and the inherited thrombophilias: does an association merit consideration? PMID- 19300048 TI - Two cases of 18 F-FDG PET/CT findings in acinar cell carcinoma of the pancreas. AB - The patients consisted of a 60-year-old woman and a 72-year-old man with no significant symptoms, who were both referred to the hospital due to the presence of large pancreatic tumors. They underwent F-18 FDG PET/CT and subsequently a pancreaticoduodenectomy and acinar cell carcinoma in the pancreas was proven histopathologically. In one case, the tumor consisted of a solid component presenting intense FDG uptake and necrotic tissue. In another case, the tumor consisted of cystic and papillary components presenting with weak FDG uptake. This report thus documents 2 cases of acinar cell carcinoma that showed contrasting histopathologic and F-18 FDG PET/CT findings. PMID- 19300049 TI - Thallium-201 and I-123 beta-methyl iodophenyl-pentadecanoic acid dual isotope single photon emission computed tomography for evaluating reperfusion injury after successful reperfusion therapy. AB - We report a reperfusion injury after rotational coronary atherectomy (RA) in a 66 year-old man with coronary artery disease. Submaximal exercise with thallium-201 single photon emission computed tomography (SPECT) imaging before reperfusion showed partially reversible perfusion defects in the apex and reversible perfusion defects in the anteroseptal area. Thallium-201 and I-123 beta-methyl iodophenyl-pentadecanoic acid (BMIPP) dual isotope SPECT was performed 5 days before and 1 hour after RA, and 1 month after RA. SPECT images at 1 hour after recovery of no reflow phenomenon after RA revealed enlargement of the defect sizes on thallium-201 and BMIPP uptakes in the anteroseptal area including the apex compared with those before RA. The defect size of thallium-201 uptake was progressively improved on 5 hour delayed redistribution imaging and 1 month after reperfusion compared with that of BMIPP uptake. In conclusion, the changes for the worse of thallium-201 uptake and fatty acid metabolism immediately after the no reflow phenomenon may indicate an injured membrane integrity with altered myocardial metabolism rather than myocardial ischemia. Thallium-201 and I-123 BMIPP dual isotope SPECT is useful for evaluating reperfusion injury after successful reperfusion therapy in a patient with acute coronary syndrome. PMID- 19300050 TI - Dual-time-point F-18 FDG PET/CT for evaluation of intrathoracic lymph nodes in patients with non-small cell lung cancer. AB - PURPOSE: The aim of this study was to evaluate the diagnostic capacity of F-18 fluorodeoxyglucose dual-time-point (DTP) positron emission tomography (PET)/computed tomography (CT) for intrathoracic lymph node (LN) metastases in patients with nonsmall cell lung cancer (NSCLC). MATERIALS AND METHODS: Thirty four patients had DTP PET/CT, with 60 minutes and 2-hour scans (n=19, NSCLC; n=15, benign pulmonary disease). LN diagnoses were confirmed by surgery or clinical follow-up (n=14, metastatic LNs; n=45, nonmetastatic LNs; n=39, inflammatory LNs). RESULTS: The maximum standardized uptake value (SUVmax) in the metastatic group was significantly higher than those in the nonmetastatic and inflammatory groups on both early- and delayed-phase imaging (each P<0.0001). The retention index (RI) of SUVmax (RI-SUVmax) in the metastatic group was significantly higher than in the nonmetastatic (P=0.0008) and inflammatory groups (P=0.0074). No significant difference was found between SUVmax values of the nonmetastatic and inflammatory groups on early- (P=0.6461) or delayed-phase (P=0.6913), or between RI-SUVmax values of the nonmetastatic and inflammatory groups (P=0.5717). For early-phase SUVmax, the cut-off value for highest accuracy with metastatic LNs was 3.61, yielding a sensitivity of 86.67% and a specificity of 88.00%. For delayed-phase SUVmax, the cut-off value was 4.00, yielding a sensitivity of 91.6% and specificity of 92.9%. For RI-SUVmax, the cut-off value was 20.91%, yielding a sensitivity of 73.6% and specificity of 75.9%. CONCLUSIONS: DTP PET/CT with a semiquantitative technique may improve diagnostic capacity for nodal staging of NSCLC. PMID- 19300051 TI - Primary testicular lymphoma involving the spermatic cord and gonadal vein. AB - We report a case of a 70-year-old man with a history of prostatic adenocarcinoma and a 3-month history of right hemiscrotal swelling. The patient underwent a CT scan, scrotal ultrasound, and F-18 FDG-PET scan to evaluate for metastatic prostate cancer. The CT scan demonstrated an ill-defined soft-tissue mass extending along the right gonadal vein. Scrotal ultrasound revealed a heterogeneous right testicular mass. The F-18 FDG-PET scan demonstrated intense hypermetabolic activity along the course of the right gonadal vein extending to the right hemiscrotum. Subsequent right radical orchiectomy and pathologic examination revealed a B-cell lymphoma, infiltrating the testicular parenchyma, spermatic cord, gonadal vessels, and adjacent soft-tissues. Lymphoma or other tumors rarely infiltrate the spermatic cord, and have only very rarely been demonstrated on PET imaging. PMID- 19300052 TI - F-18 fluorodeoxyglucose positron emission tomography/computed tomography imaging in left thigh intramuscular myxoma. AB - We report F-18 FDG PET-CT appearance of biopsy-proven intramuscular myxoma (IM) in a 69-year-old woman who presented with a left thigh mass. The tumor appears as a well-defined, hypodense mass with thin septation on CT images and mild uptake on PET images. IMs are rare benign myxoid tumors characterized by a paucity of cells, diminished vascularity, and minimal mitotic figures. They most often present as slow-growing, painless, deep-seated intramuscular masses. The mainstay of treatment is surgical resection. PMID- 19300053 TI - Functional and morphological imaging of extraadrenal myelolipoma. AB - Myelolipoma is a rare, hormonally inactive, benign tumor consisting of fat and hematopoietic cells. It is often encountered in the adrenal gland. Less frequently, it may occur in extraadrenal sites. Although myelolipoma is well defined by criteria of morphologic imaging, functional characteristics may be crucial to exclude malignancy.We present a case of histologically proven extraadrenal myelolipoma which was successfully diagnosed by bone marrow scintigraphy, using a monoclonal antibody directed against myeloid elements. PMID- 19300054 TI - Intense F-18 FDG accumulation in idiopathic tumoral calcinosis. AB - An elderly Japanese woman presented to our hospital with a 5-month history of a growing mass on her right cheek. She did not complain of local pain, headache, hearing difficulty, or tinnitus. CT and MRI showed a dense calcified mass and F 18 fluorodeoxyglucose (FDG) PET demonstrated intense accumulation in the mass. Resection of the mass was performed and it was histologically diagnosed as idiopathic tumoral calcinosis. PMID- 19300055 TI - FDG PET/CT delayed diuretic imaging technique for differentiating invasive pelvic cancer. AB - A 72-year-old woman was diagnosed as having a International Federation of Gynecology and Obstetrics (FIGO II) stage cervical cancer with diffuse uterus corpus invasion without evidence of rectal or urinary bladder invasion as determined by endoscopic examinations. Recently, 2 sharp elevations of serum tumor marker squamous cell carcinoma (SCC) were noted while she was receiving radiation therapy. Therefore, a PET/CT scan was performed using fluorodeoxyglucose (FDG) to restage the cervical cancer. FDG-avid cervical tumor was shown to extend into the anterior uterine wall by use of a diuretic imaging technique, which lowered tracer activity in the bladder and improved visualization of lower pelvic structures, which otherwise might have been obscured by urine-FDG activity. PMID- 19300056 TI - Lung sequestration and Pott disease masquerading as primary lung cancer with bone metastases on FDG PET/CT. AB - The chest x-ray from a 37-year-old man with a history of back pain showed a mass in the left lower lung, which prompted an FDG PET/CT study to evaluate the nature of the mass and possible metastases. The images revealed peripherally increased FDG activity in the left lower lung mass. In addition, there was intense FDG activity in 2 adjacent thoracic vertebrae on PET images corresponding to the regions of bone destruction on the concurrent CT. Therefore, a possible diagnosis of lung carcinoma with bone metastases was suggested. However, subsequent tests demonstrated that the left lung mass was in fact a lung sequestration, whereas the spinal lesions were due to Pott disease (tuberculous spondylitis). PMID- 19300057 TI - Thromboembolism in pulmonary artery sarcoma. AB - Pulmonary artery sarcoma, although rare, must be considered in the differential diagnosis of pulmonary thromboembolism. Clinically and radiologically, it may imitate pulmonary embolism, making diagnosis difficult and delaying treatment. Patients often have no symptom resolution despite therapeutic anticoagulation. Visualization of filling defects within a pulmonary artery on contrast-enhanced CT cannot reliably differentiate between pulmonary thromboembolism and malignant lesions like leiomyosarcoma. FDG PET-CT offers the potential for identification of malignant lesions. The authors report a case with pulmonary artery thromboembolism due to thrombi formed on a pulmonary artery leiomyosarcoma. Integrated FDG PET-CT showed no FDG-uptake along the major part of the filling defect within the right main pulmonary artery suggesting blood clot and increased uptake along the posterior wall of the right main pulmonary artery and the left lower lobar artery suggesting malignancy. PMID- 19300058 TI - Elevated FDG uptake in right middle segmental bronchus impacted with foreign body. AB - A 62-year-old man with recurrent episodes of coughing underwent a whole-body FDG PET/CT scan to evaluate possible lung malignancy. The PET images showed increased FDG uptake in the right side of the bronchus corresponding to a foreign body on the concurrent CT images. The foreign body was subsequently removed through bronchofiberscopy and was proven to be a fishbone. PMID- 19300059 TI - Lung parasite ova granuloma mimicking lung malignancy on FDG PET-CT. AB - A 60-year-old lady with mild fever, blood tinged sputum, and a left upper lung mass had whole body F-18 2-deoxy-D-glucose (FDG) positron emission tomography (PET)-computer tomography (CT) for cancer staging. The PET-CT imaging revealed FDG accumulation in the left upper lung lesion. The histopathology of the left upper lobe lung resection showed a parasite tunnel with necrosis, parasite ova, and foreign body giant cells. After surgery, eosinophiles in the peripheral blood dropped from 20% of white cell counts before operation to 5.5% at 7 weeks postoperation. PMID- 19300060 TI - Findings of subdural hematoma on Tc-99m-TRODAT-1 SPECT. AB - Chronic subdural hematoma (SDH) is difficult to diagnose by clinical manifestations only. Nonspecific neurologic symptoms and signs may lead physicians to make other diagnoses. Although head trauma is the most common cause, it may be considered insignificant or omitted due to its minor initial manifestations. We present a patient with an incidental finding of SDH on Tc-99m TRODAT-1 SPECT that was originally done for evaluation of his Parkinsonism. This reminds us to be vigilant about other possible coincidental findings on routine examinations. PMID- 19300061 TI - Utility of SPECT/CT as an adjunct to planar whole body I-131 imaging: liver metastasis from papillary thyroid cancer. AB - One of the major limitations of planar I-131 imaging is its lack of anatomic precision. SPECT/CT offers the benefit of precise anatomic localization that planar imaging lacks. Whether for confirmation of physiologic uptake or true pathology, SPECT/CT has an important role to play in clarifying equivocal findings. We present a case of papillary thyroid cancer metastatic to the liver, a relatively rare scenario. SPECT/CT allowed definitive lesion characterization at the time of the patient's visit to the nuclear medicine department. PMID- 19300062 TI - Physiologic FDG uptake in the ovary together with an abdominal wall leiomyoma mimicking metastasizing ovarian cancer on PET/CT imaging. AB - F-18 fluorodeoxyglucose (FDG)-PET-CT is increasingly used in the management of patients with ovarian cancer. However, there is a considerable overlap in the imaging features of malignant and benign ovarian lesions because physiological FDG-uptake in the ovaries can occur, depending on the menstrual cycle in premenopausal women.We present a case of FDG uptake in the ovary co-occurring with an intensely FDG active tumor of the abdominal wall in a 44-year-old woman. PET-CT findings, together with medical history, raised the suspicion for a metastasizing ovarian cancer. However, histologic examination demonstrated benign findings, namely abdominal wall leiomyoma and an ovarian follicular cyst. PMID- 19300063 TI - Diffuse skeletal muscle F-18 fluorodeoxyglucose uptake in advanced primary muscle non-Hodgkin's lymphoma. AB - A 65-year-old man newly diagnosed with primary muscular non-Hodgkin's lymphoma underwent staging F-18 Fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT). This demonstrated striking FDG uptake involving multiple skeletal muscles. Although a variety of factors may result in diffuse skeletal muscle FDG uptake, the heterogeneity and degree of FDG muscle activity, concomitant muscle enlargement, and clinical picture represent an unusual case of advanced primary muscular lymphoma. PMID- 19300064 TI - F-18 FDG PET/CT can help differentiate SAPHO syndrome from suspected metastatic bone disease. AB - A 20-year-old woman underwent magnetic resonance imaging of the spine for back pain, which demonstrated a solitary lesion in the T2 vertebral body suspicious for a metastasis. Subsequent computed tomography and bone scintigraphy identified several further sites of bony involvement but no primary malignancy. The patient was referred for F-18 FDG PET/CT which demonstrated intense FDG activity in the sternoclavicular and sacroiliac joints, in addition to the known thoracic vertebral body involvement suggesting spondyloarthropathy rather than malignant disease. During the course of investigations, the patient developed palmoplantar pustulosis leading to a diagnosis of SAPHO syndrome. PMID- 19300065 TI - Current readings in nuclear medicine. PMID- 19300066 TI - Critical Care Nursing Quarterly: Realities and myths in critical care practice. Foreword. PMID- 19300067 TI - Threading needles in the dark: the effect of the physical work environment on nursing practice. AB - Frequently, the most critical calculations, considerations, and preparations for patient care and medication administration are made in noisy, dimly lit, and chaotic areas of the nursing unit. Healthcare has begun to recognize the impact of the physical work environment plays in the ability of humans to perform reliably and safely. This article reviews the draft guidelines recently released by the United States Pharmacopeia for public comment for the physical environment to promote safe medication administration. PMID- 19300069 TI - Photography as an adjunct in pressure ulcer documentation. AB - Documentation of pressure ulcers requires accuracy and thorough information in order for providers to prescribe treatment. Frequently, documentation among providers is inconsistent and can lead to a delay in proper treatment of the ulcer. Photography, used as an adjunct in documentation, can alleviate the inconsistencies and improve the care of pressure ulcers. PMID- 19300070 TI - Reducing the failure risk of interdisciplinary healthcare teams. AB - Interdisciplinary healthcare teams have become the new model for patient care delivery in today's complex healthcare environment of increased specialization and disciplinarity of healthcare providers. Research to support the efficacy of this model has been problematic because of poorly conceptualized interdisciplinary teams as well as methodological problems conducting the research. Findings from organizational studies and healthcare interdisciplinary research can be integrated into a theory-based model to design an educational workshop to develop team members. Development of team members is proposed as a means to enhance collaboration among team members and therefore reduce the risk of team failure. PMID- 19300071 TI - The use of mirrors in critical care nursing. AB - There is no known literature to guide the critical care nurse in the use of mirrors in patient care. This article explores how the author came to believe that mirrors were essential to nursing practice. Misconceptions and assumptions concerning mirrors are explored. A framework that conceptually explores the experience of viewing self in the mirror from the perspective of a person is presented. Five situations in which the mirror may be used in critical care are discussed. This article does not present an authoritative view on mirrors in nursing; rather, it is an invitation to dialogue about a unique element in the environment that may be used to enhance nursing care. PMID- 19300072 TI - Going around in circles: is this the best practice for preparing the skin? AB - Hospital-acquired infections, which include bloodstream infections and surgical site infections, result in high rates of morbidity and mortality in the United States annually. Proper aseptic care of the skin prior to any skin breach is of paramount importance to reduce these outcomes. The application of the most appropriate skin preparation solution is significant but possibly not as important as the technique employed to apply the solution itself. Historically, concentric circles were the method of choice taught to nurses prior to any venipuncture. More recently, the back-and-forth friction method is being promoted. There is no evidence to support either method, yet effective reduction of infections is occurring. It is the intent of this article to address concerns for hospital-acquired infections and offer evidence-based suggestions to improve outcomes, as one method of skin preparation demonstrates greater efficacy. PMID- 19300073 TI - EBP partners: doctoral students and practicing clinicians bridging the theory practice gap. AB - The theory-practice gap is one component of the barriers to implementing evidence based practice. Texas Woman's University College of Nursing and Presbyterian Hospital of Denton joined forces to bridge this gap, allowing doctoral students to provide educational offerings to practicing nurses. Through a Graduate Assistance in Areas of National Need grant, doctoral students completed supervised teaching practicums at the hospital, assisting the hospital to implement evidence-based practice with the ultimate goal of applying for Magnet status. The hospital benefited from the addition of research expertise and mentoring for the staff members. The students benefited from the opportunities to teach and to share knowledge with these clinical experts. This program has been successful in meeting the needs of both parties. Recommendations are given for other institutions interested in establishing similar relationships. PMID- 19300074 TI - Acute heart failure: too sick for discharge teaching? AB - Most patients with heart failure (HF) respond within a matter of hours to days to available medical treatments. Nursing's current challenge in HF inpatient care is their short length of stay and content dense patient education needs. Only 54% of US hospitalized HF patients received all HF-1 mandated discharge education components. By using nursing evidence and adult learning principles, we can transform HF-1 topic descriptions into a "workable" plan for our newly stable HF patients. Pragmatically viewed, we need to turn the 5 key areas upside down to meet our HF patient's needs during early hospitalization: (1) recognizing symptoms, (2) pacing rest and exercise, (3) daily weights, (4) restricting sodium and fluids, and (5) managing medications. This "organizing" issue is important to our success and costly to all those who accepted the published order as prescriptive for their video, audio, and printed discharge education materials. PMID- 19300075 TI - Reexamining fatigue: implications for nursing practice. AB - A growing body of scientific evidence demonstrates negative effects of fatigue on human performance. Nursing practice encompasses many tasks that require optimal performance. Fatigue can be the result of a multitude of contributing causes. Nurses and nursing leaders must be aware of the causes and effects of fatigue and ensure that its impact is considered where staffing and patient safety intersect. PMID- 19300076 TI - Falls aren't us: state of the science. AB - PURPOSE: This article reviews the scientific health and medical literature on accidental falls and fall prevention modalities specifically directed to the hospitalized elderly population over a 15-year period. METHODS: Electronic searches of databases include CINAHL, Cochrane Library, Dissertation Abstracts, ERIC, MEDLINE, MeSH, PubMed PEDro, Ulrich's, and Web of Science. Key words and controlled subject headings used include accidental falls, fall prevention, fall risk factors, hospitalized elderly, fall incidence and rates, and environmental and patient safety. Boolean operators were utilized. Search limits include English languages, human subjects, older adult population, clinical trials, and meta-analyses. RESULTS: Wide array and multitude of papers were accessed. Analyses of the various documents from classical seminal works to the current technological studies were performed. CONCLUSION: Fall prevention modalities may facilitate achievement of the ninth goal of The Joint Commission namely, to "reduce the risk of patient harm resulting form falls" and achievement of Healthy People 2010 15th goal namely, to "reduce death from falls." PMID- 19300077 TI - Simulation in healthcare and nursing: state of the science. AB - The face of nursing education and continuing education as we currently know it is changing at a phenomenal pace. The state of the science of simulation in nursing and healthcare exist on a continuum of various types of simulation. Several driving forces have influenced the development and diffusion of the science of simulation. Today's consumers of education have higher expectations for curricula. Healthcare is evolving with fewer resources and more complex roles. Advancing technology is creating a dependence on simulation as a teaching and learning strategy. Society in general, in the form of both public and professional organizations, supports simulation as a viable teaching and learning strategy. Current use and future research will decide the fate of simulation and if it will become embedded in nursing and healthcare education. PMID- 19300078 TI - Teaching staff nurses the CAM-ICU for delirium screening. AB - The confusion assessment method for the intensive care unit (CAM-ICU) is a tool for screening for delirium in ventilated patients that with proper training can be administered quickly by staff nurses in the ICU. Unrecognized delirium can have a range of negative consequences, and in the elderly patients, it may be the first sign of an acute illness that, if left untreated, could result in death. Appropriate and early recognition is therefore imperative. Training staff to use the CAM-ICU requires not only a basic understanding of delirium and a firm orientation to the tool's features but also some preparatory decisions about tool usage and a defined approach to integrate the tool into the physical assessment process. Preparatory decisions include (1) how the tool will be used, (2) defining the process for identifying and recording baseline mental status, and (3) defining how documentation will occur. On the basis of the experience of teaching this tool to staff nurses, a 6-step process is explicated to facilitate integration: (1) putting it in context, (2) defining the features, (3) talking about tough cases, (4) doing the assessment, (5) documenting the assessment, and (6) continuing to discuss. PMID- 19300079 TI - Methicillin resistant Staphylococcus aureus (MRSA) prevention through facility wide culture change. AB - Hospitals across the United States and around the world are fighting the escalation of drug-resistant infections. In response to this costly and life threatening problem, facilities are engaging in vigorous programs to prevent its occurrence and spread. This article presents one hospital's fight against methicillin-resistant Staphylococcus aureus by implementing a facility-wide program aimed at changing and standardizing the culture within the hospital. PMID- 19300080 TI - Perceived met and unmet needs of family members of patients in the pediatric intensive care unit. AB - Extensive research has been conducted over the years with a general focus on the family members of adult patients in the critical care setting. The resulting recognition of the profound impact that the family has on the patients' outcomes has led to greater focus on the direct assessment and identification of perceived met and unmet needs of family members. In contrast to the adult patients needs, this descriptive, exploratory pilot study consisted of 20 family members who had pediatric patients in the pediatric intensive care unit. Families completed a 2 part instrument: the Demographic Data Questionnaire and the Needs Met Inventory. Data were analyzed and reported the top 10 always met/usually met needs and the top 10 never met/sometimes met needs in order of importance during the first 24 to 36 hours after admission of the pediatric patient. The overall items under the subscale assurance ranked the highest as perceived needs always met/usually met. The overall items under the subscale support ranked the lowest as perceived needs never met/sometimes met. PMID- 19300081 TI - Interferon and heart disease. AB - Heart disease is the leading cause of morbidity and mortality in the United States. Atherosclerosis is the major cause of heart disease. It is a chronic inflammatory response. Many factors contribute to this chronic inflammation. This review looks at the role of interferon gamma in atherosclerosis. PMID- 19300082 TI - The essence of life purpose. AB - Life purpose is an important thread of critical care nursing. However, no consensus exists for a definition of life purpose. In addition, ambiguity prevails regarding the manner in which life purpose is incorporated into nursing practice and research. Therefore, through a conceptual synthesis process, this article aims to clarify the essence of life purpose with relevance to health and critical care nursing today. The outcome of the conceptual synthesis is an operational definition to be used in future nursing research. Information was obtained from a literature search of scholarly articles using (1) searches of electronic databases of literature about life purpose and (2) research studies addressing conceptual, substantive, and methodological domains. Topics consisted of the philosophical underpinnings of life purpose, its attributes, definitions, and theoretical frameworks, along with differences in theories and empirical support. Finally, emerging from this process, the article culminates with a proposed conceptual definition of life purpose, which may be applied broadly to older adults in various critical care settings. PMID- 19300085 TI - Correction of coagulopathy in warfarin associated cerebral hemorrhage. AB - PURPOSE OF REVIEW: Warfarin is the most commonly used oral anticoagulant. Intracranial hemorrhage is the most serious complication of anticoagulation and the anticoagulant effect of warfarin has to be urgently reversed in this situation. Traditional methods of reversal of the anticoagulant effect of warfarin involving the use of vitamin K and fresh frozen plasma are slow and relatively ineffective and there is a need for alternative treatment approaches. RECENT FINDINGS: Agents such as prothrombin complex conjugates and recombinant activated factor VII are being increasingly used to emergently correct warfarin associated coagulopathy. Over the last decade, several small case series have suggested that these agents may lead to more rapid correction of the INR, however, improved clinical outcome is yet to be proven. A recent small prospective trial has also demonstrated the safety of a prothrombin complex conjugate and its efficacy in rapidly correcting an elevated INR in these patients. SUMMARY: There is a need for well designed randomized clinical trials aimed at evaluating the efficacy of these agents in improving the outcome of patients with anticoagulant associated intracranial hemorrhage. PMID- 19300086 TI - What is the best way to feed patients with pancreatitis? AB - PURPOSE OF REVIEW: Patients with acute pancreatitis have traditionally been treated with 'bowel rest'. Recent data, however, suggest that this approach may be associated with increased morbidity and mortality. This paper reviews evolving concepts in the nutritional management of patients with acute pancreatitis. RECENT FINDINGS: Both experimental and clinical data strongly support the concept that enteral nutrition started within 24 h of admission to hospital reduces complications, length of hospital stay and mortality in patients with acute pancreatitis. Clinical trials suggest that both gastric and jejunal tube feeding is well tolerated in patients with severe pancreatitis. Although there is limited data for the optimal type of enteral feed, a semielemental formula with omega-3 fatty acids is recommended. On the basis of current evidence, immune modulating formulas with added arginine and probiotics are not recommended. SUMMARY: Nutritional support should be viewed as an active therapeutic intervention that improves the outcome of patients with acute pancreatitis. Enteral nutrition should begin within 24 h after admission and following the initial period of volume resuscitation and control of nausea and pain. Patients with mild acute pancreatitis should be started on a low-fat oral diet. In patients with severe acute pancreatitis, enteral nutrition may be provided by the gastric or jejunal route. PMID- 19300087 TI - Probiotics in the intensive care unit. AB - PURPOSE OF REVIEW: To examine current knowledge regarding the utility of probiotics in a variety of medical conditions afflicting critically ill patients in the intensive care unit (ICU). RECENT FINDINGS: Recent experimental and clinical studies have furthered our understanding regarding the use of probiotic therapy across various clinical conditions. These disorders include antibiotic associated diarrhea, Clostridium difficile-associated diarrhea, acute pancreatitis, ventilator-associated pneumonia, and sepsis among others. Although each of these conditions is germane to ICU patients, few studies have specifically studied this vulnerable population. The current data supporting the use of probiotics in the treatment of these different clinical conditions consist mostly of the results of small, single-center trials with varying quality of research design. Although recent studies have also generally demonstrated favorable results, one well designed study in severe pancreatitis found increased mortality with probiotic therapy. These results emphasize the need for improved data regarding mechanisms of action as well as rigorous attention to safety monitoring during the execution of probiotic clinical trials. SUMMARY: Data supporting the use of probiotics in different clinical conditions are variable in scope and quality. Large, well designed, randomized, multicenter trials are needed to better define the role and safety of probiotics in critically ill patients. PMID- 19300088 TI - Current world literature. PMID- 19300090 TI - Pro's and con's of the early use of insulin in the management of type 2 diabetes: a clinical evaluation. AB - PURPOSE OF REVIEW: Recently, there have been increasing calls for insulin to be used as the initial treatment of type 2 diabetes, and if not then, soon after its onset. The underlying reason given is that insulin will slow down the apoptosis of pancreatic beta-cells, which is increased in type 2 diabetes. This review will examine the clinical evidence supporting this recommendation. RECENT FINDINGS: Several observational studies in which newly diagnosed type 2 diabetic patients are intensively treated for a short time with insulin, which is then stopped, have shown that approximately half of these patients retain good control without pharmacological therapy for up to a year. However, HbA1c levels in patients who have to be started on oral antidiabetic drugs are similar to the values in those who do not. HbA1c levels are similar in patients randomized to initial therapy with insulin or oral antidiabetic drug. There is no clinical evidence yet for an effect of insulin on beta-cell apoptosis. SUMMARY: The primary goal is to achieve and maintain HbA1c levels of less than 7.0%. Given the extra demands on both patients and physicians when starting insulin compared with oral antidiabetic drug and the many subsequent years in which patients have diabetes, the arguments for using insulin initially, or in patients who have achieved the target HbA1c level, are not convincing. However, as soon as oral antidiabetic drug therapy cannot meet this goal, insulin must be introduced. PMID- 19300091 TI - HbA1c: how do we measure it and what does it mean? AB - PURPOSE OF REVIEW: Description of recent developments in the standardization of HbA1c measurement and interpretation of HbA1c results. RECENT FINDINGS: HbA1c is extensively used in the management of patients with diabetes. The two major schemes to standardize HbA1c produce values that differ substantially. A prospective, multinational study revealed a linear correlation between HbA1c and average blood glucose. Some, but not all, assay methods are able to accurately measure HbA1c in individuals with common hemoglobin variants. SUMMARY: Progress in standardization of methods for HbA1c measurement has significantly reduced variation among different methods. The improved accuracy could allow HbA1c to be used for screening and diagnosis of diabetes. A consensus document recommends that HbA1c be reported in both NGSP (%) and IFCC (mmol/mol) units. HbA1c results can be translated into estimated average glucose (eAG), which could be reported in addition to HbA1c. PMID- 19300092 TI - Bone metabolism in type 2 diabetes and role of thiazolidinediones. AB - PURPOSE OF REVIEW: To assess bone turnover both at the biochemical and organ level in patients with type 2 diabetes (T2D) and the effects of the thiazolidinediones. RECENT FINDINGS: Studies have shown a decreased bone formation and an increased risk of fractures in patients with T2D. Changes in bone strength from glycation of collagen and negative calcium balance from calcium loss in the urine due to hyperglycaemia may also be seen. The thiazolidinediones affect bone turnover by increasing the formation of adipocytes instead of the bone-forming osteoblasts from the common mesenchymal stem cell. A decreased bone formation with decreased bone density and an increased risk of fractures has been observed among users of thiazolidinediones. Differences exist between type 1 diabetes (T1D) and T2D with a much higher increase in the risk of hip fractures in T1D than in T2D compared with the general population. The often higher body mass index in T2D than in T1D appears to explain some of the differences in risk of fractures. SUMMARY: Diabetes is a hitherto overlooked risk factor for osteoporosis and fractures. Thiazolidinediones may increase risk of fractures and should not be used by patients at risk of fractures. More research is needed. PMID- 19300093 TI - Diabetes management for intense exercise. AB - PURPOSE OF REVIEW: People with type 1 diabetes want to enjoy the benefits of sport and exercise, but management of diabetes in this context is complex. An understanding of the physiology of exercise in health, and particularly the control of fuel mobilization and metabolism, gives an idea of problems that may arise in managing diabetes for sport and exercise. RECENT FINDINGS: Exercise is complicated both by hypoglycaemia and hyperglycaemia in particular circumstances. Recent data demonstrate both early and late hypoglycaemia associated with endurance exercise and also give new insights into fuel use during exercise in diabetes. These data also provide potential explanations for the reduction in maximal exercise capacity sometimes observed in people with diabetes, although it should be noted that this observation is by no means universal. SUMMARY: Advances in the understanding of exercise physiology allow the development of management strategies that aim to help athletes with diabetes achieve appropriate metabolic control during exercise. These metabolic strategies, coupled with observations from each athlete's own experience, give a basis for individualized advice that will help athletes with diabetes to fulfil their full potential. PMID- 19300094 TI - Current Opinion in Endocrinology, Diabetes & Obesity. Current world literature. PMID- 19300096 TI - The current status of neuroimaging for epilepsy. AB - PURPOSE OF REVIEW: Neuroimaging research continues apace and is being applied to further understanding of the epilepsies, and improve clinical management. RECENT FINDINGS: Structural imaging has become more sensitive with developments of MRI hardware, acquisition and postprocessing methods. Tractography is being used to define critical pathways prior to surgery. Functional MRI for language lateralization is now a clinical tool. PET studies with specific ligands reveal neurochemical changes associated with specific epilepsy syndromes. SUMMARY: MRI at 3T with FLAIR and multiple channel coils identifies and clarifies relevant abnormalities in 20% of patients with previously unremarkable scans. Voxel-based analysis of diffusion scans may identify abnormalities in group comparisons. Identification of relevant abnormalities using voxel-based methods in individual patients requires a careful balance of sensitivity and specificity, and has a 10 30% yield. The PROPELLER sequence improves the detail of hippocampal anatomy and correlation with histological slices shows the pathological basis of MRI signal changes. Tractography has shown the connections of the language cortex and visualizes specific tracts. Electroencephalograms with simultaneous functional MRI and perfusion have shown that perfusion changes are a major determinant of changes in blood-oxygen-level-dependent signal. Functional MRI of language and memory are becoming used as a predictor of deficits as a result of temporal lobe resection.Increased uptake of the PET tracer 11C-alpha-methyl tryptophan shows promise for localizing epileptogenic malformations of cortical development. Abnormalities of 5HT-1A receptor ligands have been reported in temporal lobe epilepsy, with controversial association with depression. Dopamine uptake abnormalities have been noted in autosomal dominant nocturnal frontal lobe epilepsy. PMID- 19300097 TI - Current world literature. PMID- 19300098 TI - Relationship between ventricular morphology and aqueductal cerebrospinal fluid flow in healthy and communicating hydrocephalus. AB - OBJECTIVES: Differences in the magnitude of cerebrospinal fluid (CSF) volumetric flow through the cerebral aqueduct between healthy and hydrocephalic patients have been previously reported. However it is not clear whether this is directly related to the pathophysiology or secondary to altered ventricular morphology and hydrodynamics. This work aims to determine the role of anatomic and hydrodynamic factors in modulating the magnitude of CSF flow through the aqueduct. MATERIALS AND METHODS: Twenty subjects (10 healthy and 10 patients with communicating hydrocephalus of different causes) were studied by MRI. Scans included T1 weighted 3D anatomic imaging and velocity-encoded cine phase-contrast scans of transcranial blood and CSF flows as well as CSF flow through the aqueduct. Anatomic MR data were used for quantitation of ventricular volumes, third ventricular width, and gray and white brain tissue volumes. Velocity-encoded imaging was used for quantitation of aqueductal and cervical CSF stroke volumes (SV), aqueductal lumen area, and systolic maximal intracranial volume change. Because data from normal and hydrocephalic patients were aggregated, a battery of statistical methods that accounted for the group effects were used. Partial correlation was used to determine which of these parameters were most significantly associated with aqueductal stroke volume (ASV). Multiple linear regression analyses were employed to identify anatomic and hydrodynamic models with the least amount of variables that are significant predictors of ASV. Finally, the association between the magnitude of ASV and the aqueductal lumen area, and its implication on the CSF flow dynamic characteristics and aqueductal pressure difference was established. RESULTS: Using partial correlations, 5 of the 6 anatomic parameters and none of the hydrodynamic parameters and brain tissue volume were found to be statistically significant. The highest partial correlations were with the total ventricular volume (r = 0.838) and third ventricle width (r = 0.811). These parameters were also found to be significant predictors of ASV in the multiple linear regression analyses with third ventricle volume and group effects as insignificant predictors (F = 28.08, P < 0.0001, R = 0.85). On the other hand, both cervical CSF SV and maximal ICVC were found to be weak predictors of ASV with group effects as the only significant variable of the hydrodynamic model (F = 4.18, P = 0.023, R = 0.33). A combined anatomic hydrodynamic model including the predictive variables of the anatomic model and the ICVC provides the strongest coefficient of determination (R = 0.873). Pearson correlation analysis revealed a very strong relationship between ASV and the aqueductal lumen area (r = 0.947). CONCLUSIONS: Aqueductal CSF flow is strongly correlated with ventricular morphology, especially with the total ventricular volume and the third ventricle width, but not with the tested hydrodynamic parameters. In addition, ASV is linearly correlated with aqueductal lumen area, suggesting that the aqueductal CSF flow characteristics can be explained by oscillating pressure differences on the order of less than 0.01 mmHg. These findings may explain why a standalone ASV is a poor diagnostic marker and an insensitive indicator of shunt outcome in idiopathic normal pressure hydrocephalus. PMID- 19300099 TI - Brain tumor enhancement in magnetic resonance imaging at 3 tesla: intraindividual comparison of two high relaxivity macromolecular contrast media with a standard extracellular gd-chelate in a rat brain tumor model. AB - OBJECTIVES: The aim of this study was to evaluate lesion enhancement (LE) and contrast-to-noise ratio (CNR) properties of P846, a new intermediate sized, high relaxivity Gd-based contrast agent at 3 Tesla in a rat brain glioma model, and to compare this contrast agent with a high relaxivity, macromolecular compound (P792), and a standard extracellular Gd-chelate (Gd-DOTA). MATERIALS AND METHODS: Seven rats with experimental induced brain glioma were evaluated using 3 different contrast agents, with each MR examination separated by at least 24 hours. The time between injections assured sufficient clearance of the agent from the tumor, before the next examination. P792 (Gadomelitol, Guerbet, France) and P846 (a new compound from Guerbet Research) are macromolecular and high relaxivity contrast agents with no protein binding, and were compared with the extracellular agent Gd-DOTA (Dotarem, Guerbet, France). T1w gradient echo sequences (TR/TE 200 milliseconds/7.38 milliseconds, flip angle = 90 degrees , acquisition time: 1:42 minutes:sec, voxel size: 0.2 x 0.2 x 2.0 mm, FOV = 40 mm, acquisition matrix: 256 x 256) were acquired before and at 5 consecutive time points after each intravenous contrast injection in the identical slice orientation, using a dedicated 4-channel head array animal coil. The order of contrast media injection was randomized, with however Gd-DOTA used either as the first or second contrast agent. Contrast agent dose was adjusted to compensate for the different T1 relaxivities of the 3 agents. Signal-to-noise ratio, CNR, and LE were evaluated using region-of-interest analysis. A veterinary histopathologist confirmed the presence of a glioma in each subject, after completion of the imaging study. RESULTS: P792 showed significantly less LE as compared with Gd-DOTA within the first 7 minutes after contrast agent injection (P < 0.05) with, however, reaching comparable LE values at 9 minutes after injection (P = 0.07). However, P792 provided significantly less CNR as compared with Gd-DOTA (P < 0.05) for all examination time points. P846 provided comparable but persistent LE as compared with Gd-DOTA (P < 0.05) and demonstrated significantly greater LE and CNR when compared with P792 (P < 0.05). No statistically significant differences between CNR values for Gd-DOTA and P846 were noted for all examination time points (P < 0.05), with P846 administered at one-fourth the dose as compared with Gd-DOTA. CONCLUSION: The intravascular contrast medium P792 showed significantly less LE and CNR in comparison to Gd DOTA and P846, suggesting that it does not show marked extravasation from tumor neocapillaries and does not significantly cross the disrupted blood brain-barrier in this rat glioma model. In distinction, P846 provides comparable enhancement properties at a field strength of 3 Tesla to the extracellular contrast agent Gd DOTA, using the adjusted dose, suggesting that it crosses the disrupted blood brain-barrier and tumor capillaries, most likely based on the decreased molecular weight as compared with P792. At the same time, the high relaxivity of this compound allows for decreasing the injected gadolinium dose by a factor of 4 whereas providing comparable enhancement properties when compared with a standard extracellular Gd-chelate (Gd-DOTA) at a dose of 0.1 mmol/kg body weight. PMID- 19300100 TI - Ultra-low-dose, time-resolved contrast-enhanced magnetic resonance angiography of the carotid arteries at 3.0 tesla. AB - PURPOSE: To determine whether time-resolved magnetic resonance angiography (TR MRA) with ultra-low-dose gadolinium chelate (1.5-3.0 mL) can reliably detect or rule out hemodynamically significant disease in the carotid-vertebral artery territory. MATERIALS AND METHODS: Hundred consecutive patients (62 women, 38 men, mean age = 56.6 years) underwent both TR-MRA and standard high-resolution contrast-enhanced magnetic resonance angiography (CE-MRA), having been randomized to 1 of 2 groups; group A receiving a contrast dose of 1.5 mL for TR-MRA and group B receiving 3.0 mL. For scoring purposes the arterial system was divided into 21 segments. All TR-MRA and CE-MRA studies were blindly assessed by 2 radiologists for overall image quality, segmental arterial visualization, grading of arterial stenosis/occlusion, and incidence and severity of artifact. TR-MRA findings were directly compared with those of the corresponding CE-MRA examinations. RESULTS: Group A TR-MRA studies were of significantly inferior overall image quality compared with those of the corresponding CE-MRA examinations (P = 0.01 for both observers). In group B, overall image quality was similar for TR-MRA and single-phase CE-MRA examinations. On a segmental basis, a higher number of "insufficient quality" segments were identified in group A TR MRA studies than in group B. A similar reduction in the incidence of artifacts was observed for group B relative to group A TR-MRA studies. Both groups A and B TR-MRA studies were of high specificity, negative predictive values, and accuracy (>97%). CONCLUSION: Ultra-low dose TR-MRA may be performed with 3 mL of gadolinium chelate with preservation of overall image quality and arterial segmental visualization relative to single phase CE-MRA, whereas a 1.5 mL contrast dose is associated with more suboptimal studies. Nonetheless, even at doses as low as 1.5 mL, TR-MRA can exclude arterial stenosis or occlusion. PMID- 19300101 TI - Proinflammatory cytokine changes in clinically stable, virologically suppressed, HIV-infected patients switching from protease inhibitors to abacavir. PMID- 19300102 TI - The relationship between prolonged antiretroviral therapy and cryptogenic liver disease. PMID- 19300103 TI - Discordant immunologic and virologic responses to antiretroviral therapy and associated mortality in a large treatment program in Rwanda. PMID- 19300104 TI - Further insights into the 24-h blood pressure profile by home blood pressure monitoring: the issue of morning hypertension. PMID- 19300105 TI - Prehypertension in China. PMID- 19300106 TI - Cardio-renal organ damage and cardiovascular outcomes in hypertension. PMID- 19300107 TI - Angiogenesis inhibition and preeclampsia. PMID- 19300108 TI - Remodelling of small cerebral arteries in human hypertension: structural and functional alterations. PMID- 19300109 TI - Prehypertension and risk of cardiovascular disease in Chinese adults. AB - OBJECTIVE: To examine the excess risks associated with prehypertension and absolute benefit of treatment. METHODS: We conducted a prospective cohort study in 169 871 Chinese adults aged 40 years and older. Data on blood pressure and covariables were obtained at a baseline examination in 1991 and follow-up evaluation was conducted in 1999-2000. RESULTS: Compared with normotension (<120/80 mmHg), prehypertension (120-139/80-89 mmHg) was significantly associated with an increased relative risk (95% confidence interval) of cardiovascular disease (CVD) incidence 1.34 (1.27, 1.42) and mortality 1.22 (1.15, 1.30), coronary disease incidence 1.32 (1.16, 1.50) and mortality 1.47 (1.23, 1.75), and stroke incidence 1.72 (1.59, 1.86) and mortality 1.67 (1.50, 1.86). The population-attributable risk associated with prehypertension was 10.6 and 7.1% for CVD incidence and mortality, 9.9 and 13.9% for coronary disease incidence and mortality, and 19.9 and 18.7% for stroke incidence and mortality. The optimal 5 year number-needed-to-treat to prevent a CVD event or death was 53 and 185, 17 and 51, and eight and 22 for prehypertension, stage 1 hypertension, and stage 2 hypertension, respectively. The optimal number-needed-to-treat to prevent a CVD event was significantly smaller in prehypertension patients with a history of CVD or diabetes (34 for incidence and 44 for mortality) compared with those without (115 for incidence and 352 for mortality). CONCLUSION: Prehypertension was related to an increased risk of CVD. Treatment of prehypertension among patients with a history of CVD or diabetes was as beneficial as treatment of stage 1 hypertensive patients without a history of CVD or diabetes. PMID- 19300110 TI - Influence of exercise at lower and higher intensity on blood pressure and cardiovascular risk factors at older age. AB - OBJECTIVE: It is not well known which exercise intensity is needed to obtain blood pressure reductions in response to endurance training. We therefore compared the effect of training at lower and higher intensity on blood pressure, and, in addition on other cardiovascular risk factors, in at least 55-year-old sedentary men and women. METHODS: We used a randomized crossover design comprising three 10-week periods. In the first and third periods, participants exercised at, respectively, lower and higher intensity (33 and 66% of heart rate reserve) in random order, with a sedentary period in between. Training programmes were identical except for intensity and were performed three times, 1 h per week. Thirty-nine (18 men) out of 48 randomized participants completed the study; age averaged 59 years. RESULTS: The change of aerobic power from baseline to the end of each period was more pronounced (P < 0.05) with higher intensity (+3.70 ml/kg min; P < 0.001) than with lower intensity training (+2.31 ml/kg min; P < 0.001). Systolic blood pressures at rest and during submaximal exercise were reduced with both intensities (P < 0.01), whereas diastolic office blood pressure was significantly reduced after higher intensity only (P < 0.01). There were no significant differences in blood pressure reduction between intensities. Ambulatory blood pressure remained unchanged after training. Only higher intensity training reduced weight (-1.09 kg; P < 0.001), body fat (-0.85%; P < 0.001), plasma triglycerides (-0.17 mmol/l; P < 0.05) and oxidized low-density lipoprotein (-5.92 U/l; P < 0.01). CONCLUSION: Higher and lower intensity training reduces systolic office and exercise blood pressure to a similar extent, but does not alter ambulatory blood pressure; only higher intensity training favourably affects anthropometric characteristics and blood lipids. PMID- 19300111 TI - Day-by-day variability of spontaneous baroreflex sensitivity measurements: implications for their reliability in clinical and research applications. AB - OBJECTIVE: To assess day-by-day variability of spontaneous baroreflex sensitivity (BRS) measurements, providing implications for their reliability in clinical and research studies. METHODS: Forty-four healthy volunteers and 57 patients with previous myocardial infarction (MI) underwent an 8 + 8 min (spontaneous + paced breathing) recording of ECG and noninvasive arterial pressure on two consecutive days. BRS was computed according to the sequence method (BRS_seq), the original and modified transfer function method (BRS_TF and BRS_TF(mod)) and the alpha method (BRS_alphaLF and BRS_alphaHF). Absolute and relative reliability were assessed by the 95% limits of random variation (LoV) and by the intraclass correlation coefficient (ICC), respectively. The sample size needed to detect a clinically relevant change was also estimated. RESULTS: In healthy volunteers during spontaneous breathing, BRS_seq, BRS_TF, BRS_alphaLF and BRS_alphaHF could not be measured in 18, 3, 2 and 2% of recordings, respectively. By definition, BRS_TF(mod) could always be measured. The 95% LoV indicated that individual day by-day changes may range from -50% to +101% for BRS_TF(mod) (best case) and from 58% to +135% for BRS_alphaHF (worst case). The ICC ranged from 0.70 (BRS_seq) to 0.76 (BRS_TF(mod)). The sample size varied from 56 (BRS_TF(mod)) to 80 (BRS_seq). In MI patients, measurability was lower whereas reliability indexes were similar. Results during paced breathing were similar. CONCLUSION: Day-by-day variability should be taken into account when using spontaneous BRS measurements to detect treatment effects in individual patients. The observed substantial to good relative reliability, as assessed by the ICC, indicates that spontaneous BRS measurements are suitable to detect differences between individuals, which is a prerequisite for proper diagnosis and prognosis. PMID- 19300112 TI - Altered structure of small cerebral arteries in patients with essential hypertension. AB - OBJECTIVE: Structural alterations in the microcirculation may be considered an important mechanism of organ damage. An increased media-to-lumen ratio of subcutaneous small resistance arteries has been demonstrated to predict the development of cardiocerebrovascular events in hypertensive patients. Alterations in the structure of small cerebral arteries have been demonstrated in animal models of experimental or genetic hypertension. However, no evaluation with reliable techniques has ever been performed in humans. DESIGN AND METHODS: Twenty eight participants were included in the present study: they were 13 hypertensive patients and 15 normotensive individuals. All participants underwent a neurosurgical intervention for benign or malign tumors. A small portion of morphologically normal cerebral tissue was excised from surgical samples and examined. Cerebral small resistance arteries (relaxed diameter around 200 mum) were dissected and mounted on an isometric and isobaric myograph, and the tunica media to internal lumen ratio was measured. In addition, cerebral cortical microvessel density (MVD) was also evaluated. The tissue was sectioned and stained for CD31, and MVD was measured with an automated image analyzer (percentage of area stained). Blood pressure values were evaluated, before surgical intervention, by standard sphygmomanometry. RESULTS: M/L was significantly greater and MVD significantly lower in hypertensive patients than that in normotensive individuals. No difference between groups in collagen content or mechanical properties of cerebral small arteries was observed. CONCLUSION: Our results indicate that structural alterations of small cerebral vessels are present in hypertensive patients compared with normotensive individuals, similar to those previously observed in subcutaneous small arteries. PMID- 19300113 TI - Detection of silent cerebrovascular disease refines risk stratification of hypertensive patients. AB - OBJECTIVE: Detection of preclinical hypertension-related cardiorenal damage has been recommended in the identification of patients most at risk of cardiovascular complications. The inclusion of silent cerebrovascular disease (SCD) as an additional marker of hypertensive organ involvement might improve risk stratification. METHODS: In 192 hypertensive patients (98 men) without a history of cardiovascular and cerebrovascular disease, a mean age of 51.6 +/- 12.3 years and untreated office blood pressure levels of 170 +/- 23/104 +/- 12 mmHg, we obtained detailed information on preclinical cardiac (left ventricular hypertrophy), renal (microalbuminuria, impaired kidney function or both) and cerebrovascular damage (white matter hyperintensities, infarcts, microbleeds or all), and estimated the associated cardiovascular risk on the basis of the presence of common cardiovascular risk factors. RESULTS: Hypertensive target organ damage involved the heart in 41 (21%), the kidneys in 50 (26%) and the brain in 84 (44%) participants. When considering only patients with demonstrable cardiac, renal damage or both (n = 72), 42 participants (58%) had also SCD. Of the remaining 120 participants without cardiorenal damage, 42 (35%) had brain damage. In other words, half of all patients with SCD were classified as having no target-organ (i.e., cardiorenal) involvement. The cardiovascular risk score of patients without cardiorenal but with brain damage was significantly higher than that of participants without any organ involvement (37 +/- 11 versus 27 +/- 11, P < 0.001), and similar to the risk score of those with cardiorenal damage (38 +/- 14, P > 0.05). CONCLUSION: These data suggest that SCD should be recognized as an additional, independent and prognostically relevant marker of preclinical hypertensive target-organ damage. PMID- 19300114 TI - Impact of supportive measures on drug adherence in patients with essential hypertension treated with valsartan: the randomized, open-label, parallel group study VALIDATE. AB - OBJECTIVE: The majority of treated hypertensive patients do not achieve target blood pressure (BP) levels of less than 140/90 mmHg. One key reason is inadequate adherence with the prescribed drug regimen. Dosing regimens are either not executed as prescribed (noncompliance) or patients stop taking the medication (nonpersistence). It has been demonstrated that adherence with angiotensin receptor antagonists such as valsartan is high due to the tolerability profile of this drug class. The present study was designed to evaluate whether drug adherence could further be improved by the use of supportive measures. DESIGN AND METHODS: Twenty-eight centers were randomized to provide pharmacological treatment with or without a set of supportive measures (e.g. structured physician patient interaction, printed information about hypertension, reminder stickers, 24 h timer, and home BP measurement device). Two hundred and two patients with grade 1 hypertension (BP at baseline 149.8 +/- 6.2/93.9 +/- 4.4 mmHg) who were either newly diagnosed or who had not been treated for at least 1 year were included in this trial. All patients entered the 34-week treatment phase with valsartan 160 mg daily. Titration to valsartan 160 mg/hydrochlorothiazide 12.5 mg was allowed if necessary. Drug adherence was assessed by electronic monitors (Medication Event Monitoring System). RESULTS: Patients treated with a valsartan based therapy receiving supportive measures as compared with the standard care group demonstrated an initially higher level of adherence with a maximum absolute difference of 7.8% (P = 0.041). This difference did not persist over the observation period but faded with time. In parallel, execution of the dosing regimen (compliance) was also improved in the intervention group during the early months of treatment but this effect also disappeared by the end of the observation period. In contrast, persistence in the two groups slowly but continuously separated over time. Estimated absolute difference in persistence at the end of the 34-weeks study between the two groups was 7.6% (95.9 vs. 88.3%) reflecting a 66% lower hazard of discontinuation in the intervention group (P = 0.073). BP control improved more in patients with the supportive measures. CONCLUSION: Drug adherence improved initially with the use of supportive measures. However, this effect faded with time mainly because of the short-lived improvement in the quality of execution (compliance) achieved. In contrast, a longer lasting effect of the chosen supportive measures on persistence could be demonstrated, which, however, at least under the conditions of the present study, did not translate into a persistent improvement of medication adherence. PMID- 19300116 TI - Interesting findings in the VALERIA trial. PMID- 19300117 TI - Prognostic significance of flow-mediated dilatation of the brachial artery in hypertensive patients; possible role of central blood pressure. PMID- 19300118 TI - Pharmacotherapy for functional gastrointestinal disorders in children. AB - Therapy of pediatric functional gastrointestinal disorders is best done within the context of a multidisciplinary biopsychosocial approach. Pharmacotherapy is often sought by patients and families who hope to find a "pill" that will lead to rapid symptom relief. Yet, there is only scant published evidence for the efficacy of a variety of medical interventions in childhood functional abdominal pain and irritable bowel syndrome. This article reviews the pediatric studies that have addressed pharmacotherapy in children with pain predominant functional gastrointestinal disorders. PMID- 19300119 TI - Novel and alternative therapies for childhood constipation. AB - Constipation is a common and distressing pediatric problem. Even if prevalence of this disorder has increased, treatment of constipation in children is still largely based on clinical experience rather than on evidence-based studies. The currently recommended treatment includes education, disimpaction, and maintenance therapy, consisting of dietary changes, behavioral modification, and use of laxatives. Among osmotic agents, polyethylene glycol solutions appear to be the first-line drug to use in pediatrics. Although these measures are effective in the majority of children, a sizable proportion needs long-term therapy. Chronicity may significantly interfere with the child's emotional growth and development. Advances in the understanding of the gastrointestinal enteric nervous system and epithelial function have led to the development of new classes of drugs. These include substances that bind to serotonin receptors or are chloride channel activators. Further studies are needed to assess the benefits and the risks of these new drugs. PMID- 19300120 TI - Probiotics and functional gastrointestinal disorders in children. AB - Chronic constipation is one of the most frequent complaints in childhood. Although there is evidence that gastrointestinal flora is important in gut motility, there is little evidence that gut flora is abnormal in constipation. Lactobacilli and bifidobacteria increase stool frequency and decrease consistency in normal individuals. But, according to several reviews, the evidence of probiotics for efficacy in constipation is limited. Fiber supplements, lactose free diets, and lactobacillus supplementation are effective in the management of children with recurrent abdominal pain and irritable bowel syndrome. Several studies with Lactobacillus GG in children showed negative results in children with chronic constipation. Because Bifidobacterium animalis DN-173 010 has been shown to be effective in adults with constipation-predominant irritable bowel syndrome this study should also be performed in a well-designed large placebo controlled trial in children with constipation. PMID- 19300121 TI - Third ESPGHAN Capri Meeting, "Colonic Diseases: Problems and Progress", in Capri, Italy, April 2-4, 2009. Introduction. PMID- 19300122 TI - Advances in understanding colonic function. AB - The colon is an organ of conservation that salvages water, electrolytes, and energy. The organization of colonic function is determined by the roles played by the luminal flora, the function of the different mucosal epithelial cell types, immunocompetent cells, and the neuromusculature. These different components of the colon interact with one another and with the colonic flora, and different areas of the colon serve different functions. In the normal adult during the course of a day the colon absorbs approximately 1.5 L of fluid, but under the influence of aldosterone increases up to 5 to 6 L. Diarrhoea occurs when secretion exceeds absorptive processes by either small intestinal secretion overwhelming colonic salvage or salvage being impaired by reduced colonic absorption or increased colonic secretion. PMID- 19300123 TI - Evolving concepts of the pathogenesis of irritable bowel syndrome: to treat the brain or the gut? AB - Recent in-depth studies of irritable bowel syndrome (IBS) that assessed multiple physiological endpoints in large patient cohorts confirm evidence of abnormal motility, sensation, and psychosocial disturbances. However, the proportion with hypersensitivity has dropped from the time of the original claim that it is a "biological marker" of IBS. Discomfort thresholds in male and female IBS patients normalize over time, whereas IBS symptoms persist, and increased colonic sensitivity in IBS is strongly influenced by a psychological tendency to report pain and urge rather than increased neurosensory sensitivity. The objectives of this article are to review the classical pathophysiology of IBS, the putative roles of infection, inflammation, and bacterial flora; consider mimics of IBS; assess the evidence that IBS is a serotonergic disease; evaluate recent advances in membrane biology and neuroscience related to IBS; consider genetic association with IBS and its endophenotype; and discuss whether to treat the gut or the brain. PMID- 19300124 TI - Bacterial mucosa cross-talk and pathophysiology of inflammation. AB - The intestinal mucosa harbors a complexly organized immune system, which accomplishes important, partially contradictory surveillance and protective functions. It has to protect the host from potential external (microbial and nonmicrobial) aggression, whereas at the same time avoiding an inflammatory reaction toward harmless antigenic structures of microbial (commensal microflora) or alimentary origin. Professional sentinels (of the innate immune system) are in close and interactive contact with the intestinal microflora, which plays via this exchange an important role during the postnatal period in the maturation of the intestinal immune system. The innate immune system is a key regulator of local and systemic immune responses, including the adaptive immune system. Distinct signals induce tolerance to specific antigens presented by intestinal dendritic cells, whereas proinflammatory mediators easily switch these tolerogenic responses toward protective or destructive inflammatory reactions. Given the intense exchange between the intestinal microflora and the mucosal immune system, a permanent but actively controlled and downregulated physiological inflammation characterizes the intestinal mucosa. Therefore, it is easily conceivable that perturbation of the intestinal microflora or immune control mechanisms can initiate or contribute to pathological and potentially chronic inflammatory reactions, such as seen in inflammatory bowel diseases. PMID- 19300125 TI - Risks and benefits of biologic therapy for IBD. PMID- 19300126 TI - Intestinal mucus barrier in normal and inflamed colon. AB - Intestinal bacteria are usually regarded as harmless commensals in healthy intestine but are pathogens, if these invade the tissues. The mucus gel separates the luminal bacteria from the epithelial surface throughout the colon in healthy individuals. This viscoelastic mucus gel is protective against adhesion and invasion by microorganisms, bacterial toxins, and antigens. The mucus viscosity increases progressively toward the distal colon, separating bacteria selectively in the proximal colon and completely in the distal colon. Mucus in normal subjects is usually intact and devoid of bacteria, by contrast mucus barrier is broken and penetrated by bacteria and inflammatory cells in patients with colonic inflammation. The immune inclusion hypothesis postulates that the host mucosa maintains an adherent bacterial biofilm that develops immune tolerance with specific immune mechanisms. The bacterial biofilm, growing in the mucus matrix, would prevent contact of pathogenic bacteria with the intestinal mucosal wall. On the contrary, recent evidence indicates that bacteria are absent in mucus from healthy individuals and present in mucus from patients with inflammatory bowel disease. In inflammatory bowel disease alteration in the types of mucins or, alternatively, the altered mucus layer as a response to inflammation contributes to the underlying pathology by affecting the mucus barrier function. PMID- 19300127 TI - Changes in intestinal microflora in obesity: cause or consequence? AB - Obesity and the associated metabolic disorders, such as diabetes and metabolic syndrome, have become major public health issues worldwide. Obesity results from a positive energy balance and is associated with decreased microbial diversity in the human gut with lower levels of Bacteroidetes. However, whether changes in the gut microbiota are a cause or consequence in obesity remains to be definitively proven. Experiments using germ-free mice have begun to reveal some mechanisms by which the gut microbiota may affect the development of obesity. PMID- 19300128 TI - Colon in acute intestinal infection. AB - The colon is actively implicated in intestinal infections not only as a target of enteric pathogens and their products but also as a target organ for treatment. In the presence of diarrhea, both of osmotic and secretory nature, the colon reacts with homeostatic mechanisms to increase ion absorption. These mechanisms can be effectively exploited to decrease fluid discharge. A model of intestinal infections using rotavirus (RV) in colonic cells was set up and used to define a dual model of secretory and osmotic diarrhea in sequence. Using this model, antidiarrheal drugs were tested, namely zinc and the enkephalinase inhibitor racecadotril. Zinc was able to decrease the enterotoxic activity responsible for secretory diarrhea. It also inhibited the cytotoxic effect of RV. The mechanism of zinc was related at least in part to the activation of MAPK activity, but also a direct antiviral effect was observed. Racecadotril showed a potent and selective inhibition of active secretion, being particularly effective in the first phase of RV diarrhea. The use of drugs active at the colonic level, therefore, offers effective options to treat intestinal infections in childhood. In addition, the colon is the natural site of colonic microflora, a target of probiotic therapy, which is the first line of approach recommended by the European Society for Paediatric Gastroenterology, Hepatology and Nutrition to treat infectious diarrhea. PMID- 19300129 TI - Clostridium difficile infections: emerging epidemiology and new treatments. AB - Clostridium difficile infections (CDIs) are increasingly a cause of morbidity and mortality. Although the pathogenicity of C difficile is based on toxin A and B expression, new hypervirulent strains express novel virulence factors. The etiology of recent increases in prevalence and severity of disease is not clearly explained by known mechanisms. New at-risk groups include children (without prior antibiotic exposure) and patients with inflammatory bowel disease. Vancomycin has an important role in the treatment of CDIs, but does not prevent recurrence. Other therapies for recurrent disease include antibiotics, probiotics, and immunotherapy. Novel therapies are under investigation for recurrent and resistant CDI. PMID- 19300130 TI - Role of the colon in short bowel syndrome and intestinal transplantation. AB - Colon is a crucial partner for small intestinal adaptation and function in patients who underwent extensive small intestinal resection or transplantation. This short review deals with the different properties and roles of the colon in these settings, involving fluid and electrolytes absorption, absorption of medium chain triglycerides, and production of short-chain fatty acids for malabsorbed energy salvage. The colon may adapt after small intestinal resection, whereas it hosts the most important part of the intestinal microbiota, which plays a crucial role in intestinal function and health. Also, colon may be responsible for D lactic acidosis as well, as it can be injured by noninfectious colitis. Finally, the relevance of a simultaneous colon grafting is discussed as it is occasionally considered in specific patients requiring intestinal transplantation. PMID- 19300131 TI - Short- and long-term complications of colectomy. PMID- 19300132 TI - Screening and surveillance recommendations for pediatric gastrointestinal polyposis syndromes. AB - Inherited polyposis syndromes are relatively rare disorders in pediatric gastroenterology practice, even in busy academic settings. It is important, however, for pediatric gastroenterologists to be aware of the serious health risks for children and their families affected by these disorders. The diagnosis of a polyp syndrome is often made in the first or second decade of life, long before the risk of gastrointestinal neoplasia. Pediatric gastroenterologists must be prepared then to offer families predictive genetic screening as well as endoscopic surveillance when appropriately indicated. The current overview is designed to provide general guidelines and, whenever possible, evidence-based recommendations for genetic testing, endoscopic surveillance and other screening approaches for children with inherited gastrointestinal polyposis syndromes. In this presentation, the focus is on screening for neoplastic change and complications in the gastrointestinal tract. It is important to understand that extraintestinal cancers are frequent in some of these disorders and the reader is referred to other authoritative sources for additional information about comprehensive health screening outside the gastrointestinal system. PMID- 19300133 TI - Congenital anorectal malformations: anything new? AB - Management of anorectal malformations, especially that of proximal and more severe forms have developed significantly in the last 25 years. Echography and magnetic resonance imaging have improved the diagnostic accuracy, especially in terms of associated anomalies that are the main cause of fatalities among patients with anorectal malformations. Animal models, novel molecular biological techniques and knock-out gene models have widened our knowledge on aetiology and pathogenesis of these anomalies. From early 1980s, the posterior sagittal anorectoplasty originally designed by deVries and Pena has systematized the surgical management of anorectal anomalies leading to improved functional outcomes. Surgical mini-invasive techniques are increasingly applied to the management of these anomalies, although their role is as yet not clearly established. Long-term functional follow data are also available today indicating a reasonable favourable faecal and urinary control in the majority of patients with anorectal malformations. For the patients with less favourable functional outcome there are bowel and bladder management options that provide sufficient social continence. PMID- 19300134 TI - Nutritional challenge in pseudo-obstruction: the bridge between motility and nutrition. AB - Chronic intestinal pseudo-obstruction is a severe syndrome characterized by a profound derangement of the intestinal propulsive motility that resembles mechanical obstruction, in the absence of any mechanical obstruction. This syndrome represents one of the main causes of intestinal failure and is characterized by impairment of physical growth and development as well as by a high rate of morbidity and mortality. It may be idiopathic or secondary to a variety of diseases. Most cases are sporadic, even though familial forms with either dominant or recessive autosomal inheritance have been described. Based on histological features intestinal pseudo-obstruction is classified into 3 main groups: neuropathies, mesenchymopathies, and myopathies, according to the predominant involvement of enteric neurones, interstitial cells of Cajal, and smooth muscle cells, respectively. Treatment of intestinal pseudo-obstruction involves nutritional, pharmacological, and surgical therapies, but it is often frustrating and does not change the natural course in the majority of cases. The nutritional management has a crucial importance in pediatric age and involves the administration of special formulae, the enteral delivery of nutrients, by a nasogastric tube, percutaneous gastrostomy, or jejunostomy. In the most severe cases, parenteral nutrition becomes mandatory in order to satisfy nutritional requirements and manage appropriately obstructive episodes. PMID- 19300135 TI - Nutritional aspects in inflammatory bowel disease. AB - Nutrition plays a role in inflammatory bowel disease (IBD) primarily in prevention and treatment of malnutrition and growth failure. Furthermore, in Crohn disease (CD), nutrition can induce remission, maintain remission, and prevent relapse. Malnutrition is common in IBD and the mechanisms involved include decreased food intake, malabsorption, increased nutrient loss, increased energy requirements, and drug-nutrient interactions. At the time of diagnosis, up to 85% of pediatric patients with CD and 65% of those with ulcerative colitis (UC) have weight loss. Growth failure occurs in 15% to 40% of children with IBD and is less common in UC compared with CD, both at diagnosis and during follow up. In CD, nutritional therapy with enteral formulas induces remission at a rate comparable with that achieved with steroids. In adults with CD, limited information suggests that enteral nutrition (EN) may play a role in maintenance of remission. In children with CD colitis, one study suggested that children without colitis respond better to EN than children with colitis, and another study found no such difference but reported a trend toward earlier relapse in those with isolated colonic involvement. Finally, nutrition may play a role in IBD via the possible protective effect of breastfeeding against UC and CD. In summary, although only CD may benefit from nutrition as primary therapy for remission induction and possibly maintenance of remission, nutrition plays an important role in the prevention and treatment of malnutrition in IBD, and may have a protective role, via the effect of breast-feeding on disease occurrence. PMID- 19300136 TI - Colon in food allergy. AB - The colon is a site of allergic reaction to food proteins. The most common manifestation is allergic proctocolitis. Although frequent in infancy, it may also affect older children. The diagnosis of allergic proctocolitis is mainly based on clinical and histological grounds, but there is a risk of overdiagnosis; challenge is strongly recommended to avoid unnecessary and expensive formula or changes in maternal diet that may discourage continuation of breast-feeding. The benign clinical course and the spontaneous resolution in most infants suggest the need for further prospective studies to validate markers that allow the identification of those children needing a dietary approach. It remains to be assessed whether lymphoid nodular hyperplasia of the colon is a manifestation of food allergy. Finally, allergic proctitis may present clinically with refractory constipation; the true prevalence of food allergy-related constipation remains to be assessed. PMID- 19300137 TI - New frontiers in the treatment of Hirschsprung disease. AB - BACKGROUND: Hirschsprung disease (HSCR) is the most common identifiable developmental disorder of the enteric nervous system (ENS), characterized by a failure of its formation in a variable segment of distal bowel. Currently available surgical therapies for HSCR, although lifesaving, are associated with an unsatisfactory long-term prognosis for many, highlighting the need for newer curative therapies. On the basis of our understanding of ENS development, 1 potential therapeutic tool relates to the cells that gave rise to the ENS during embryogenesis. Data from initial animal studies suggested that such "ENS stem cells" persist within postnatal gut, from which they can be harvested and used to replenish absent nerves within experimental models of gut aganglionosis. More recently, it has been shown that ENS stem cells can also be isolated from human postnatal gut. The early human studies, however, used full-thickness gut tissue obtained at surgery to generate ENS stem cells, which in clinical terms poses practical problems. In an effort to directly address such obstacles our latest research has shown that ENS stem cells can also be harvested from gut mucosal biopsies obtained at routine endoscopy. Such gut mucosal ENS stem cells are capable of appropriately and fully colonizing aganglionic gut tissue, including from HSCR, to generate components of a mature and functional ENS. CONCLUSIONS: Current evidence suggests that transplantation of ENS stem cells, sourced from easily accessible postnatal gut, could be a viable alternative treatment for HSCR and ultimately for a number of other congenital or acquired ENS disorders. PMID- 19300138 TI - Postinfectious irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a gastrointestinal disorder characterized by abdominal pain and changes in bowel habits, not sustained by structural changes. There is now consistent evidence indicating that IBS may be the adverse outcome of an acute episode of infectious gastroenteritis, the so-called postinfectious (PI) IBS. The infectious agents involved in the development of PI-IBS include pathogenic bacteria, parasites, and viruses. Abdominal pain and diarrhea are the most common symptoms of PI-IBS. Several studies identified a number of risk factors increasing the susceptibility for PI-IBS development. These include the virulence of the pathogen, the severity, and duration of the acute enteritis, younger age, female sex, and psychological disturbances. Several mucosal abnormalities in the colon or ileum of patients who develop PI-IBS have been described. These changes include increased mucosal permeability, an increased amount of intraepithelial lymphocytes, lamina propria T cells, and mast cells, as well as serotonin-containing enteroendocrine cells. The mediators released by these activated cells may evoke enteric nervous system responses, excite sensory afferent pathways, and induce visceral hyperalgesia. Little is known about the prognosis of PI-IBS, although it is likely better than that of nonspecific IBS. There is little evidence about a specific treatment for PI-IBS. Although probiotics and antibiotics may be promising in the prevention of PI-IBS, the efficacy of these treatments should be assessed in an ad hoc designed study. PMID- 19300139 TI - Functional nonretentive fecal incontinence in children: a frustrating and long lasting clinical entity. AB - Fecal incontinence is defined as the passage of stools in an inappropriate place at least once per month, for a minimum period of 2 months. This frustrating symptom is a source of considerable distress and embarrassment for the child and the family. According to the Rome III criteria fecal incontinence can be subdivided into constipation-associated fecal incontinence and functional nonretentive fecal incontinence. This short review mainly addresses functional nonretentive fecal incontinence in children. Definition, prevalence, pathophysiology, and recent updates on treatment and long-term follow-up of fecal incontinence are discussed. PMID- 19300140 TI - Thigh strength and activation as predictors of knee biomechanics during a drop jump task. AB - PURPOSE: To examine whether normalized quadriceps and hamstring strength would predict quadriceps and hamstring muscle activation amplitudes and whether these neuromuscular factors would predict knee kinematics and kinetics during a drop jump task. METHODS: Thirty-nine females and 39 males were measured for isometric quadriceps and hamstring strength and were instrumented to obtain surface electromyography, kinematic, and kinetic measures during the initial landing of a drop jump. Multiple linear regressions first examined the relationship between thigh strength and activation then examined whether these neuromuscular variables were predictive of hip and knee flexion excursions, knee extensor moments (KEM), and anterior knee shear forces during the deceleration phase of the drop jump. RESULTS: Females versus males produced lower normalized thigh strength and demonstrated greater quadriceps and hamstring activation amplitudes during the drop jump. Lower thigh muscle strength was a weak (males) to moderate (females) predictor of greater quadriceps activation amplitudes. However, thigh strength and activation were poor predictors of hip and knee joint excursions and KEM. Regardless of sex and thigh strength, anterior shear forces were greater in individuals who demonstrated less hip flexion and greater knee flexion excursions and greater peak quadriceps activation and internal KEM during the landing. CONCLUSIONS: Although thigh muscle strength explained some of the variance in quadriceps and hamstring activation levels as measured with surface electromyography, we failed to support the hypothesis that these neuromuscular factors are strong predictors of sagittal plane hip and knee flexion excursions or KEM. Although greater quadriceps activation amplitude was a significant predictor of greater anterior tibial shear forces, its contribution was relatively small compared with kinematic and kinetic variables. PMID- 19300141 TI - Aerobic exercise and postprandial lipemia: issues on volume and frequency of exercise. PMID- 19300143 TI - Beta-blocker and exercise capacity: an always relevant interference? PMID- 19300145 TI - High versus low concentration botulinum toxin A for benign essential blepharospasm: does dilution make a difference? AB - PURPOSE: To evaluate the efficacy and side effects of concentrated versus dilute botulinum toxin A in treating benign essential blepharospasm. METHODS: The authors performed a prospective randomized clinical trial of 16 patients with an established diagnosis of benign essential blepharospasm. Patients were randomized to receive low concentration (control, 10 U/ml) injections on one side and high concentration (experimental, 100 U/ml) injections on the other. They were surveyed on a scale of 1 to 10 regarding pain, bruising, and redness immediately after the injection. During their return visit, at an established interval of 1 to 3 months, patients were questioned regarding complications (ptosis, diplopia, tearing, and dry eye), duration of relief, and side preferred. Patients were followed over 8 months for 1 to 6 repeat injections, with the side given the higher concentration alternated at each visit. RESULTS: With 16 patients, there were a total of 42 visits and 84 observations (eyes) documented. Using the Wilcoxon rank sum test, there was a statistically significant reduction in pain scores (1.94 vs. 4.59, p < 0.001) on the experimental side versus the control side. Patient assessment revealed no significant difference in bruising, redness, complications of injection, side preference, or length of relief of symptoms. CONCLUSIONS: Compared with the control, the high concentration botulinum toxin A demonstrated a 58% reduction in perceived pain. Patients did not report a significant difference in efficacy or complications with either dilution. PMID- 19300146 TI - Full-thickness eyelid resection in the treatment of secondary ptosis. AB - PURPOSE: To describe horizontal full-thickness eyelid resection for the treatment of ptosis as related to overcorrections after levator recession for eyelid retraction in Graves disease or repeatedly failed external levator advancement. METHODS: The authors present a retrospective case study of 19 eyelids of 17 patients with refractory ptosis. Patients were selected based on history of horizontal full-thickness eyelid resection to treat ptosis secondary to over recessed Muller and levator muscles, persistent ptosis after levator advancement/resection, or persistent ptosis after internal levator advancement. Results were graded from eyelid measurements taken before and at least 6 weeks after treatment. RESULTS: All eyelids were measured preoperatively and postoperatively for the following parameters: palpebral fissure in primary gaze nasally (N), centrally (C), and temporally (T); upper eyelid margin to light reflex distance in primary gaze; upper eyelid margin to crease distance; and upper eyelid margin to fold distance. Final postoperative measurements were taken at a minimum of 2 months. Thirteen eyelids had a history of previous Muller muscle excision/levator recession, 3 eyelids had a history of previous levator advancement/resection, 2 eyelids had previous internal levator advancement, and 1 eyelid had previous cosmetic eyelid surgery. All 17 patients were women. Patients ranged in age from 41 to 63 years. All patients had cosmetically and functionally satisfactory results. CONCLUSIONS: Horizontal full-thickness eyelid resection is a unique procedure that can provide an excellent result for patients with residual ptosis with no recurrence of ptosis during an average of 2 to 3 months follow-up. PMID- 19300147 TI - Upper eyelid ptosis surgery using a preparatory ocular prosthesis. AB - PURPOSE: When an ocular prosthesis is unable to correct upper eyelid ptosis, surgery is performed with the existing prosthesis, which may be difficult and unsatisfactory. We used an alternative approach, in which the prosthesis was modified as a preparation to surgery. METHODS: This retrospective, noncomparative, interventional case series included 26 anophthalmic and 3 microphthalmic patients with unilateral, moderate to severe upper eyelid ptosis who underwent levator aponeurosis surgery via an anterior approach. Excluded were cases of congenital anophthalmos, contracted socket, and neurogenic ptosis. Before surgery, the prosthesis was modified to vertically align the pupils while neglecting the position of the upper eyelid. The outcome measures were palpebral fissure height and cosmetic appearance. RESULTS: The amount of ptosis before the procedure ranged from 1 to 6 mm (mean, 3 mm). After surgery, 24% (7 of 29) of patients received a minor adjustment to the preparatory prosthesis to improve the pupil or eyelid position. After the entire procedure, the ptosis was symmetrically corrected with the contralateral eye in 19 patients (66%), with 1 mm of residual ptosis in 6 patients (17%), and with 2 mm of residual ptosis in 4 patients (14%). No overcorrection was seen. CONCLUSION: The alternative sequence of a preparatory prosthesis before surgery provides good results in the correction of moderate to severe ptosis. PMID- 19300148 TI - Addition of dermis-fat graft to diminish cable visibility in frontalis suspension for patients with pre-existing deep superior sulci. AB - PURPOSE: To propose and demonstrate a technique modification for frontalis suspension to simultaneously address ptosis and diminish abnormalities related to deep superior sulcus defect. METHODS: Six patients underwent simultaneous frontalis suspension and placement of a superior sulcus dermis-fat graft. RESULTS: Postoperative visibility of the cables, anticipated in patients with deep superior sulci, was prevented in these 6 patients. CONCLUSIONS: Dermis-fat grafting may be added to improve cosmesis after frontalis suspension in patients at risk for postoperative cable visibility. PMID- 19300149 TI - Comparison of fibrin sealant versus suture for wound closure in Muller muscle conjunctiva resection ptosis repair. AB - PURPOSE: To compare fibrin sealant (Tisseel) versus suture for wound closure in Muller muscle-conjunctiva resection ptosis repair. METHODS: The charts of 114 patients (211 eyelids) who had undergone Muller muscle-conjunctiva resection were retrospectively reviewed. Suture versus Tisseel were used for wound closure. Preoperative and postoperative eyelid measurements, postoperative symmetry within 0.5 mm, and complications were compared. RESULTS: Muller muscle-conjunctiva resection ptosis repair was performed on 211 eyelids of 114 patients. Seventeen cases were unilateral and 97 cases were bilateral. Method of wound closure included suture (45 eyelids of 31 patients) versus Tisseel (166 eyelids of 83 patients). For the suture group, the mean preoperative MRD1 was 1.2 mm and the postoperative MRD1 was 3.0 mm; the difference was 1.9. For the Tisseel group, the mean preoperative MRD1 was 1.2 mm and the postoperative MRD1 was 3.0 mm; the difference was 1.8. The 2 groups did not differ statistically in preoperative (p = 0.97) or postoperative MRD1 values (p = 0.53), the difference (p = 0.63), or postoperative symmetry within 0.5 mm (p = 0.39). In the suture group, complications included moderate to severe pain (10%), suture granuloma (6%), corneal abrasion (3%), loose suture (3%), and persistent keratopathy (3%). We found no evidence of keratopathy attributable to the Tisseel (p = 0.0001). This difference in the prevalence of complications was statistically significant (p = 0.0001). Four patients in the suture group (13%) underwent subsequent procedures including suture granuloma removal (2) and suture removal (1); 1 patient (3%) required levator resection. Three patients in the Tisseel group (4%) subsequently underwent levator resection. CONCLUSIONS: Muller muscle-conjunctiva resection ptosis repair using fibrin sealant for wound closure offers comparable eyelid position results compared with suture. Use of Tisseel showed fewer postoperative complications and was associated with fewer subsequent surgical procedures. PMID- 19300150 TI - Clinical characterization and blepharoptosis surgery outcomes in Hispanic New Mexicans with oculopharyngeal muscular dystrophy. AB - PURPOSE: To clinically characterize blepharoptosis in Hispanic New Mexicans with oculopharyngeal muscular dystrophy and examine eyelid surgery outcomes. METHODS: A retrospective noncomparative case series and retrospective, nonrandomized, comparative interventional case series was performed on medical records from 86 patients. Main outcome measures included preoperative correlations between margin reflex distance, palpebral fissure height, levator function, and age and postoperative change in palpebral fissure height and time to reoperation for recurrent blepharoptosis after blepharoplasty, levator advancement, or frontalis sling surgery. RESULTS: Preoperative measurements between the right and left eye were symmetrical with respect to margin reflex distance, palpebral fissure height, and levator function (all p < 0.001). There were correlations between age and margin reflex distance, palpebral fissure height, and levator function (all p < or = 0.02). There was no gender difference detected with respect to age, margin reflex distance, palpebral fissure height, and levator function (p > 0.39). Eighty-three patients underwent eyelid surgery. As initial surgery, 15 underwent blepharoplasty, 17 levator advancement, and 51 frontalis suspension. Overall, 93.3% of blepharoplasty patients, 47.1% levator advancement patients, and 7.84% undergoing frontalis suspension had additional surgery for recurrent ptosis (rates differed, p < 0.001). Postoperative change for palpebral fissure height was 0.33 +/- 1.83 mm OD and 1.1 +/- 0.86 mm OS for levator advancement and 2.63 +/- 1.34 mm OD and 2.68 +/-1.47 mm OS for frontalis suspension (p = 0.03, OD and p = 0.004, OS). CONCLUSIONS: Oculopharyngeal muscular dystrophy in Hispanic New Mexicans is a symmetrical, progressive disease that affects men and women similarly. Frontalis suspension is an effective primary surgery with respect to upper eyelid elevation, need for reoperation, and time to reoperation in this patient population. PMID- 19300151 TI - Infection rates in outpatient eyelid surgery. AB - PURPOSE: The authors used the database of eyelid surgeries performed at the Phillips Eye Institute in Minneapolis, MN, to help determine the infection rate in outpatient eyelid surgery. METHODS: The authors retrospectively reviewed the reported cases of infections after all eyelid surgeries performed between January 1, 1999 and December 31, 2004 at the Phillips Eye Institute in Minneapolis, MN. RESULTS: Of a total of 2,227 patients who underwent oculoplastic procedures, there was only one reported infection (0.04%). CONCLUSIONS: The infection rate in outpatient eyelid surgery is extremely low. PMID- 19300152 TI - Selective embolization and resection of a large noninvoluting congenital hemangioma of the lower eyelid. AB - PURPOSE: To present a combined interventional radiologic and resection technique for the management of a large amblyogenic, highly vascular lesion of the lower eyelid associated with severe mechanical ectropion, conjunctival pyogenic granuloma, and recurrent, spontaneous conjunctival bleeding. METHODS: Preoperative selective embolization and coil placement were followed by resection of the tumor. RESULTS: Embolization, coil placement, and resection were uneventful with cosmetic and functional visual improvement obtained. Pathology indicated a noninvoluting congenital hemangioma. CONCLUSIONS: To the authors' knowledge, this is the first surgical technique reported in the ophthalmic literature for the selective embolization of a noninvoluting congenital hemangioma prior to its resection. Such a technique should be considered in the management of large lesions of this type. PMID- 19300153 TI - Photodynamic therapy using methyl aminolevulinate acid in eyelid basal cell carcinoma: a 5-year follow-up study. AB - PURPOSE: To evaluate retrospectively the long-term results of methyl aminolevulinate photodynamic therapy for the treatment of eyelid basal cell carcinoma. METHODS: Sixteen consecutive patients with eyelid basal cell carcinoma were treated with methyl aminolevulinate photodynamic therapy between January 2002 and April 2003. Selection criteria were tumors located at least 3 mm from the tarsus, surgery not indicated because of poor general health, and recurrences with unclear location definition. Patients were treated with an 80-J cm light emitting diode light source (632 nm) after topical application of methyl aminolevulinate cream and occlusion for 4 hours. Data were available for follow up at day 1, week 1, month 1, and every 6 months for 5 years. RESULTS: The mean number of photodynamic therapy session per patient was 3.1 (range, 2-6) Complete clinical recovery was observed after the 5-year follow-up in 13 of 16 patients (82%). Two patients did not respond at all to treatment and 1 patient presented with recurrence after 3 years of tumor-free follow-up. Tolerance of treatment was good with few side effects. CONCLUSIONS: The absence of complications, good tolerance, and a notable success rate make this nonsurgical procedure promising for the treatment of basal cell carcinoma of the eyelid in selected patients. PMID- 19300154 TI - Use of Lorenz titanium implants in orbital fracture repair. AB - PURPOSE: To present the experience and outcomes of orbital fracture repair using Lorenz titanium orbital implants in one tertiary care center. METHODS: Five-year retrospective chart review of 55 patients who underwent orbital fracture repair with Lorenz titanium orbital implants since 2002. The following data were recorded: age; gender; mechanism of injury; prior fracture repair; concomitant ocular injuries; time elapsed between injury and surgery; and preoperative and postoperative visual acuity, diplopia, extraocular motility, and Hertel exophthalmometry. All patients underwent preoperative CT. Operative data included surgical approach, type of Lorenz implant used, duration of surgery, and presence of extraocular muscle entrapment. Length of follow-up and complications were also recorded. RESULTS: Of those undergoing primary repair, diplopia in primary gaze was present in 29.3% of patients preoperatively and 0% postoperatively. Severe, moderate, and mild motility restriction in any gaze direction were 5.5%, 14.5%, and 34.5% preoperatively and decreased to 0%, 1.8%, and 7.3% postoperatively. Although only 14.3% of primary repair patients had full extraocular motility prior to surgery, this number increased to 80% after surgical repair. More than 80% of patients achieved symmetry within 1 mm by Hertel exophthalmometry. The authors did not encounter instances of implant extrusion, infection, or eyelid malposition during follow-up. CONCLUSIONS: Orbital fracture repair using Lorenz titanium orbital implants provides reliable and reproducible stabilization of orbital wall defects, while offering outstanding functional and cosmetic results comparable with other alloplastic materials. PMID- 19300155 TI - Hess area ratio and diplopia: evaluation of 30 patients undergoing surgical repair for orbital blow-out fracture. AB - PURPOSE: To determine if the Hess area ratio is effective in predicting postoperative diplopia in patients undergoing surgery for orbital blow-out fracture. METHODS: Our retrospective, interventional case series study involved 30 consecutive cases affected by orbital fractures and diplopia undergoing surgical correction within 7 days after injury. To evaluate ocular motility disturbance, we measured the involved ocular motility range by use of a manual Hess screen test before and 4 months after surgery. The percentage of Hess area ratio % was used to express the range of ocular motility in a numerical value. RESULTS: All patients with preoperative Hess area ratio >85% had no postoperative diplopia, and most patients (57%) having a preoperative Hess area ratio <65% had postoperative diplopia. When the Hess area ratio was between 65% and 85%, surgical outcomes were variable and most patients (55%) described no problematic diplopia in the peripheral visual field. CONCLUSIONS: The Hess area ratio is a useful procedure to convert Hess graphic representation in a numerical value so that Hess chart data can be compared among clinicians and used to predict surgical outcomes in patients undergoing surgery for orbital blow-out fractures. PMID- 19300156 TI - Visual outcomes in patients with paranasal mucoceles. AB - PURPOSE: To describe the ophthalmic complications and visual outcome of patients with paranasal mucoceles. METHODS: This is a retrospective case series of patients with a diagnosis of paranasal mucocele. Their medical records were examined with particular emphasis on the location of the paranasal mucoceles, presence of inflammation (mucocele vs. pyelocele), presence of optic neuropathy, and final visual outcome. RESULTS: Ten patients with an average age of 57.6 years (range, 30-71 years) were studied. Seven mucoceles were in the frontal or fronto ethmoidal sinuses, one was in the sphenoidal sinus, one was within an Onodi cell, and one was in the maxillary sinus. Four patients developed optic neuropathy, of which 2 cases were related to fronto-ethmoidal mucoceles. Four patients had pyeloceles. Two of these 4 patients had resultant loss of light perception: one case was associated with a fronto-ethmoid sinus and the other with an Onodi cell). CONCLUSIONS: Our series highlights the greater risk of optic neuropathy and poor visual outcome with sphenoid sinus and Onodi cell mucoceles because of their proximity to the optic nerve. However, fronto-ethmoidal mucoceles are not benign and may compromise vision, especially if a posterior ethmoid component is present, or if treatment is delayed and secondary infection has ensued to form a pyelocele. Management of paranasal mucoceles should be prompt, particularly when signs of inflammation are present. PMID- 19300157 TI - Enucleation with hydroxyapatite implantation versus evisceration plus scleral quadrisection and alloplastic implantation. AB - PURPOSE: To compare the motility and complications of enucleation with evisceration plus scleral quadrisection. METHODS: In a prospective study between January 2006 and June 2007, 50 patients underwent evisceration with scleral quadrisection and alloplastic implantation (group 1) and 50 patients underwent enucleation and hydroxyapatite implantation (group 2). Horizontal and vertical excursions of implants and complications such as exposure or extrusion and deep superior sulcus deformity were evaluated postoperatively. RESULTS: After a mean follow-up interval of 11.5 months for group 1 and 13.2 months for group 2, vertical measurements were approximately 20% less than horizontal measurements in both groups. The mean horizontal and vertical movements in group 2 were significantly less than in group 1 (p < 0.001). There was 1 case of small hydroxyapatite exposure in group 2 (1 in 50; 2%) and 2 cases of implant extrusion in group 1 (4%) (p = 0.50). Deep superior sulcus deformity was noted in 10 patients in group 1 (20%) and 7 patients in group 2 (14%) (p = 0.43). Analysis of covariance in both groups showed that age, gender, and follow-up interval were not predictors of movement in either direction (all p-values >0.05). CONCLUSIONS: Evisceration with scleral quadrisection and alloplatic sphere implantation can effectively substitute for a more extensive procedure such as enucleation in patients with painful blind eyes, cosmetically unacceptable blind eyes, and medically uncontrolled endophthalmitis. It provides rapidity, ease, and better implant excursion and lower cost of the implants compared with their porous counterparts. PMID- 19300158 TI - Rectangular 3-snip punctoplasty outcomes: preservation of the lacrimal pump in punctoplasty surgery. AB - PURPOSE: To report a method of rectangular 3-snip punctoplasty (2 vertical cuts at either side of the vertical canaliculus and 1 cut at the base). Outcomes are compared with the more conventional and anatomically destructive 3-snip triangular punctoplasty (1 cut at the vertical canaliculus, 1 cut at the horizontal canaliculus, and 1 cut at the base). METHODS: Retrospective review of all eyes undergoing 3-snip punctoplasty between January 1, 2003 and January 8, 2005. RESULTS: Seventy-five patients were eligible for this study. One hundred eight eyes with patent punctum and lacrimal systems prior to surgery underwent punctoplasty (49 rectangular and 59 triangular punctoplasty). Follow-up of 8 days to 2.3 years (mean, 0.68 years). Ages ranged between 16.8 and 99.8 years (mean, 69.5 years). Forty-five patients (60%) were women. There was a trend for patients to be symptomatic postoperatively despite patent punctum and lacrimal systems following triangular (10 of 59, 16.9%) than those who underwent rectangular punctoplasty (5 of 49, 10.2%) although this difference was not significant (difference in proportions = 0.05; CI: -17.635 to 17.645; p = 0.99). CONCLUSIONS: This study showed that there were more failures in eyes with triangular punctoplasty. Previous studies have shown that the punctae and canaliculi are important in normal lacrimal drainage. Rectangular punctoplasty is a simple, safe, and quick procedure that maintains the normal anatomy and physiology of the canalicular system, while maintaining a patent punctum and should be considered as a less destructive alternative to the traditional triangular punctoplasty. PMID- 19300159 TI - Normalization of conjunctival flora after dacryocystorhinostomy. AB - PURPOSE: To determine the time of normalization of conjunctival flora after successful dacryocystorhinostomy. METHODS: Forty consecutive adult patients with nasolacrimal duct obstruction and chronic dacryocystitis who had been referred for dacryocystorhinostomy were included in the study. Conjunctival cultures were obtained preoperatively from the involved and normal sides of each patient. Cultures from the lacrimal sac contents were obtained at the time of dacryocystorhinostomy. Postoperatively, conjunctival specimens were obtained from the operated side weekly until the result of the cultures was negative or the colony count was less than that of the normal side. RESULTS: The mean time of normalization was 4.5 weeks (range, 3-8 weeks). The time of normalization was significantly related to the type of organisms (p = 0.04) and the colony count (p < 0.001). The mean time of normalization was 3.5 weeks in patients with a colony count <10 and 4.93 weeks in patients with a colony count > or =10. The time of normalization was significantly related to the presence of a silicone tube (p < 0.0001). The mean time of normalization was 6.2 weeks and 3.9 weeks in patients with and without a silicone tube, respectively. CONCLUSION: A few weeks is needed for the conjunctival flora to normalize after dacryocystorhinostomy. The time of normalization is related to the type of bacteria, colony count, and presence of silicone tube. PMID- 19300160 TI - Hypotony maculopathy after eyelid massage for overcorrected blepharoptosis. AB - A 57-year-old woman with a history of left eye trabeculectomy was evaluated for gradual ipsilateral visual loss. Several months prior, she had undergone levator advancement of the left upper eyelid. For management of resulting retraction, she was instructed to "massage" her eyelid. Examination was notable for left eye visual acuity of 20/200 and an intraocular pressure of 5 mm Hg. On fundoscopic examination, the macula was edematous with multiple folds, consistent with hypotony maculopathy. Two years later, after obliteration of the bleb and placement of a Seton valve, the intraocular pressure has increased to 8 mm Hg with an acuity correctable to 20/50. Ocular hypotony may result from digital eyelid massage in patients with filtering blebs. Clinicians should bear this in mind when managing patients with eyelid retraction after blepharoptosis repair. PMID- 19300161 TI - Frontalis suspension sling using palmaris longus tendon in chronic progressive external ophthalmoplegia. AB - A 26-year-old woman referred with bilateral ptosis was diagnosed to have underlying chronic progressive external ophthalmoplegia. The authors report satisfactory result at 18-month follow-up with palmaris longus tendon used as an autologous sling material for ptosis surgery in this patient. PMID- 19300162 TI - Parotid tumor presenting with hemifacial spasm. AB - A 47-year-old man presented with right parotid swelling and a history of frequent attacks of hemifacial spasm. MRI of the brain and neck showed a mass in the right parotid gland. Fine needle aspiration biopsy of the mass revealed a pleomorphic adenoma of the parotid gland, which was confirmed after total right parotidectomy. His attacks of hemifacial spasm did not improve after surgery and 8 months postoperatively, he received botulinum toxin-A injections, which improved his symptoms. Clinicians need to be aware that patients with occult parotid tumors can present like patients with classic hemifacial spasm. PMID- 19300163 TI - Myeloma of the eyelid: a rare necrotizing lesion. AB - A 77-year-old man was referred to the authors with an impression of recurrent chalazion of the left lower eyelid. He had suffered from multiple myeloma and had undergone chemotherapy for 11 months. A 2.0 x 0.8 cm, firm, tender, oval-shaped, ulcerated mass with an accompanying black crust on its surface, and erythematous swelling around it was observed. Evaluation of an eyelid biopsy specimen revealed neoplastic plasma cells with eccentric nuclei and monoclonality for lambda light chain, which the authors report as the first case of metastatic multiple myeloma presenting as a rare necrotizing eyelid lesion. PMID- 19300164 TI - Darier disease mimicking Basal cell carcinoma of the eyelid. AB - Darier disease is a rare autosomal dominant skin disorder that infrequently has ocular manifestations. The authors describe 2 cases of patients with known Darier disease presenting with eyelid lesions resembling basal cell carcinomas. Biopsies demonstrated histopathologic changes consistent with Darier disease. To the authors' knowledge, this presentation has not been previously reported. Ophthalmologists and dermatologists should be aware of this type of presentation in Darier disease. PMID- 19300165 TI - Tarsal dermoid cyst: clinical presentation and treatment. AB - The authors present 2 cases of eyelid dermoid cyst attached to tarsus in pediatric patients. Both patients were infants who presented with a firm, nontender upper eyelid mass firmly adherent to tarsus. In both cases, the lesion was excised en bloc, and histopathology revealed a dermoid cyst. To the authors' knowledge, there are no previously reported cases of tarsal dermoid cyst. These cases demonstrate the importance of including dermoid cyst in the differential diagnosis of a tarsus-based eyelid mass. Misdiagnosis may lead to incision and curettage, resulting in spillage of cyst contents and the risk of severe inflammation and scarring. PMID- 19300166 TI - Steatocystoma simplex of the eyelid. AB - A 68-year-old woman presented with a nodular pretarsal tumor on her right upper eyelid that had slowly grown over 6 months. Excisional biopsy was performed through an eyelid crease approach, and histopathology evidenced the lesion to be steatocystoma simplex. Steatocystoma simplex is an uncommon benign cutaneous lesion thought to be a circumscribed malformation arising from the pilosebaceous duct junction, rarely reported to occur in the periorbital region. PMID- 19300167 TI - Conjunctival edema and distichiasis in association with congenital lymphedema of the lower legs. AB - An 8-year-old boy with mild congenital lymphedema of both legs was noticed to have a conjunctival lesion of the right eye since birth. Topical corticosteroid eyedrops for slight irritation did not alter the appearance of the lesion. On ocular examination, diffuse conjunctival edema in the inferotemporal and inferonasal quadrants and mild injection of the right eye was noted. There was no edema on the left eye. Three extra rows of eyelashes (congenital distichiasis) were found bilaterally on upper and lower eyelids. Related systemic anomalies included mild aortic coarctation, left ventricular hypertrophy, and simian crease on each palm. Conjunctival edema and distichiasis are important ophthalmic features of the congenital lymphedema distichiasis syndrome. PMID- 19300168 TI - Foreign body after pterygium surgery simulating an epibulbar tumor. AB - A 60-year-old woman was referred for evaluation of a rapidly growing conjunctival neoplasm after pterygium surgery. The lesion proved to be a foreign body granuloma, with exogenous material that seemed to be cotton mesh, probably used for tamponade of subconjunctival bleeding that was unintentionally left in place. PMID- 19300169 TI - Bilateral dacryoadenitis: a new addition to the spectrum of reactive arthritis? AB - Reactive arthritis is a well-known sequel to both dysenteric and urethritic illnesses, classically occurring in combination with conjunctivitis, arthritis, and/or a variety of dermatologic conditions. Dacryoadenitis has not been previously described as part of the spectrum of reactive arthritis. The authors describe a case of biopsy-proven bilateral dacryoadenitis, which developed in the setting of chlamydial urethritis. PMID- 19300170 TI - Primary orbital leiomyosarcoma. AB - A 79-year-old woman was presented with a 6-month history of painless proptosis in the left eye. On examination, there was a palpable superotemporal mass displacing the left eye inferomedially. Orbital CT revealed a heterogeneous, lobulated mass occupying the left orbital space with no bone erosion or destruction. MRI showed an extraconal, heterogenous, lobulated orbital mass in the lacrimal gland region of the left eye. The lesion was excised, and the diagnosis of leiomyosarcoma was made by histopathologic examination and immunohistochemistry. Systemic examinations were negative for a primary or a metastatic tumor. There was no evidence of tumor recurrence after 12 months of follow-up. Primary orbital leiomyosarcoma is a rare tumor that mainly occurs in older women and presents with painless proptosis. It should be considered in the differential diagnosis of superotemporal extraconal lesions. PMID- 19300171 TI - Frontal sinus osteoma presenting with orbital emphysema. AB - Osteoma is the most common neoplasm of the paranasal sinuses usually involving the frontal sinus. Osteomas are benign, but can cause serious intracranial or orbital complications. The authors report a frontal sinus osteoma with orbital emphysema in a 16-year-old boy with a history of right-eye swelling after nose blowing. CT showed a right frontal sinus bony mass. Intraoperatively, strands of mucosa extending from the frontal sinus around the periphery of the mass in the right orbit were seen. The mass was excised and the orbital roof repaired. Histopathology was consistent with osteoma. The osteoma facilitated entrance of sinus air in the orbit, resulting in recurrent orbital emphysema with nose blowing. There was no recurrence of orbital emphysema after resection. PMID- 19300172 TI - Adult orbital langerhans cell histiocytosis with frontal bone involvement. AB - A 28-year-old woman presented with a 2-week history of right upper eyelid swelling and intermittent frontal headaches. CT demonstrated an ill-defined superior right orbital mass with adjacent right frontal bone erosion and undeveloped frontal sinuses. The orbital biopsy revealed tissue strongly positive for CD1a and S100, diagnostic of Langerhans cell histiocytosis. The systemic workup was negative for multifocal lesions and for diabetes insipidus. In addition to subtotal resection, the patient was treated with a 6-month course of oral prednisone and intravenous vinblastine. PMID- 19300173 TI - Repair of isolated abortive cryptophthalmos with lower eyelid switch flap and amniotic membrane graft. AB - The surgical correction of a case of bilateral isolated abortive cryptophthalmos is described. An upper eyelid and superior fornix were successfully created in a 2-stage procedure using a switch flap from the patient's normal lower eyelid and an amniotic membrane graft. Exposure keratopathy was significantly reduced with resultant visual improvement. PMID- 19300174 TI - Optic nerve and chiasmal germinoma. AB - A 15-year-old boy presented with visual acuity of 20/200 OD and no light perception OS. The anterior segment of the left eye showed a relative afferent pupillary defect. A large (4.5 x 4.5 x 2.0 mm) infiltrative optic nerve head lesion with dilated vessels was seen OS with disc pallor OD. MRI of the brain and orbit revealed lobulated optic nerve thickening and chiasm. A biopsy revealed features consistent with germinoma and was positive for marker placental alkaline phosphatase. Systemic examination, chest x-ray, abdominal ultrasound, cerebrospinal fluid, and serology were normal. He received 27 Gy to the craniospinal region followed by a boost of 27 Gy to the left optic nerve. Eight months postirradiation, vision stabilization was achieved with 20/200 OD and light perception with inaccurate projection of rays OS. PMID- 19300175 TI - Successful treatment of nasolacrimal duct obstruction secondary to fibrous dysplasia by external dacryocystorhinostomy with mitomycin C. AB - A 60-year-old woman developed polyostotic fibrous dysplasia involving the right zygomatic and maxillary bones, which led to swelling along the right side of her nose that was associated with constant epiphora. A sac washout revealed a blocked right nasolacrimal duct, which was confirmed by CT dacryocystography. CT also showed signs of fibrous dysplasia in the maxillary and zygomatic bones in the form of bony expansion and ground-glass bone density. The patient underwent external dacryocystorhinostomy with mitomycin C and O'Donoghue tube insertion. Bone punched out while fashioning the bony nasal ostium was subjected to histopathologic examination, which confirmed the diagnosis of fibrous dysplasia. The O'Donoghue tubes were removed 8 weeks later. At 15 months postoperatively, the dacryocystorhinostomy was patent and the patient was completely relieved of her symptoms. PMID- 19300177 TI - Sponsors and investigators in food science: vicious circle or virtuous circle? PMID- 19300178 TI - Mycophenolate mofetil decreases acute rejection and may improve graft survival in renal transplant recipients when compared with azathioprine: a systematic review. AB - BACKGROUND: Mycophenolate mofetil (MMF) has increasingly replaced azathioprine (AZA) as the antimetabolite of choice in immunosuppressive protocols. Initial trials comparing MMF with AZA in patients receiving cyclosporine A sandimmune showed a clinical benefit in reducing the incidence of acute rejections. It has been questioned whether this benefit remains significant when using newer formulations of cyclosporine A (neoral) and tacrolimus. METHODS: Literature searches were performed using the Transplant Library, Cochrane library, Medline, and Embase for all randomized controlled trials directly comparing MMF with AZA in renal transplant recipients. Trials were assessed for quality using the Jadad scoring system. Trials were pooled using meta-analysis software. Confidence intervals were set at 95%. RESULTS: Nineteen relevant studies were identified, including a total of 3143 patients. MMF significantly reduces the risk of acute rejection when used in combination with any calcineurin inhibitor (relative risk 0.62, 0.55-0.87, P<0.00001). The hazard for graft loss, including death with a functioning graft, is also significantly reduced in patients treated with MMF (hazard ratio 0.76, 0.59-0.98, P=0.037). There is no significant difference in patient survival or renal transplant function between groups. Risk of adverse events, including cytomegalovirus infection, anemia, leukopenia or rates of malignancy, does not differ significantly. A greater risk of diarrhea is seen in MMF-treated patients. CONCLUSIONS: We have shown that MMF used with a calcineurin inhibitor does indeed confer a clinical benefit over AZA by reducing the risk of acute rejection and also possibly reducing graft loss. This effect is independent of whether MMF is used in combination with sandimmune, neoral or tacrolimus. PMID- 19300179 TI - Influence of immunosuppressive regimens on graft survival and secondary outcomes after kidney transplantation. AB - BACKGROUND: There have been striking changes during the last 10 years concerning the choice of calcineurin inhibitor and antimetabolite agent prescribed after kidney transplantation. METHODS: A retrospective analysis of 51,303 patients undergoing deceased-donor kidney transplantation during 1998 to 2007 was performed using multivariate regression analysis. All patients received cyclosporine A (CsA) or tacrolimus (Tac) with azathioprine (AZA) or mycophenolic acid (MPA) on an intention-to-treat basis with corticosteroids plus/minus antibody induction. Graft survival rates and secondary outcomes were analyzed. A subanalysis was performed for transplants undertaken during 2002 to 2007, in which all patients were treated with MPA plus corticosteroids and CsA or Tac. RESULTS: All-cause graft failure and death-censored graft failure to 5 years posttransplant did not differ significantly between Tac and CsA. We found no evidence in support of previous claims that MPA results in superior long-term graft survival compared with AZA treatment. At the end of year 1, Tac was associated with a lower risk for serum creatinine more than or equal to 130 mumol/L (P<0.001) and hypercholesterolemia (P<0.001) versus CsA, but a higher risk for de novo posttransplant diabetes (P<0.001). MPA treatment was associated with a lower risk of acute rejection (P<0.001) but a higher risk of hospitalization because of infection (P<0.001) versus AZA. CONCLUSIONS: Five-year graft survival in deceased-donor kidney transplant recipients is equivalent in patients receiving CsA- or Tac-based immunosuppression, and in those receiving MPA or AZA. The absence of a survival benefit with modern agents is relevant in the current cost-conscious era of prescribing. PMID- 19300180 TI - The case of azathioprine versus mycophenolate. Do different drugs really cause different transplant outcomes? PMID- 19300181 TI - The innate immune response and activation of coagulation in alpha1,3 galactosyltransferase gene-knockout xenograft recipients. AB - BACKGROUND: The role of the innate immune system in the development of thrombotic microangiopathy (TM) after alpha1,3-galactosyltransferase gene-knockout (GTKO) pig organ transplantation in primates is uncertain. METHODS: Twelve organs (nine hearts, three kidneys) from GTKO pigs were transplanted into baboons that received no immunosuppressive therapy, partial regimens, or a full regimen based on costimulation blockade. After graft failure, histologic and immunohistologic examinations were carried out. RESULTS: Graft survival of less than 1 day was prolonged to 2 to 12 days with partial regimens (acute humoral xenograft rejection) and to 5 and 8 weeks with the full regimen (TM). Clinical or laboratory features of consumptive coagulopathy occurred in 7 of 12 baboons. Immunohistochemistry demonstrated IgM, IgG, and complement deposition in most cases. Histopathology demonstrated neutrophil and macrophage infiltrates, intravascular fibrin deposition, and platelet aggregation (TM). Grafts showed expression of primate tissue factor (TF), with increased mRNA levels, and TF was also expressed on baboon macrophages/monocytes infiltrating the graft. CONCLUSIONS: Our data suggest that (1) irrespective of the presence or absence of the adaptive immune response, early or late xenograft rejection is associated with activation of the innate immune system; and (2) porcine endothelial cell activation and primate TF expression by recipient innate immune cells may both contribute to the development of TM. PMID- 19300182 TI - Detection of immunoglobulin G human leukocyte antigen-specific alloantibodies in renal transplant patients using single-antigen-beads is compromised by the presence of immunoglobulin M human leukocyte antigen-specific alloantibodies. AB - BACKGROUND: Luminex-based single-antigen human leukocyte antigen (HLA) antibody detection beads (SAB) are a major advance for the characterization of HLA specific antibodies but their clinical utility is limited unless the analysis is performed and interpreted optimally. Here, we identify problems encountered in routine monitoring of antibody levels that may give rise to misleading results, and describe how these can be overcome to provide more meaningful clinical information. METHODS AND RESULTS: HLA class I specific antibody-binding levels were determined using SAB in the sera of 42 highly sensitized patients awaiting renal transplantation. Normalization of the results against the HLA class I specific monoclonal antibody W6/32 overcame the problems caused by variation in antigen density on SAB and also suggested the presence of alloantibodies directed against multiple HLA class I epitopes of a given specificity. Routine analysis using undiluted sera gave an incomplete assessment of antibody levels. On serum dilution, three patterns of antibody binding became apparent: most sera showed a sequential reduction in immunoglobulin G (IgG) binding levels but several sera displayed antibody binding which remained unchanged (suggesting antigen saturation) or increased IgG binding on serum dilution (suggesting inhibition of IgG binding using neat serum). The presence of immunoglobulin M (IgM) HLA specific antibodies in sera correlated with inhibition of IgG antibody binding for the corresponding specificity and treatment of sera with dithiothreitol to eliminate IgM HLA-specific blocking antibodies restored maximum IgG antibody binding levels. CONCLUSION: When using SAB to monitor HLA-specific antibody binding levels, sera should be pretreated with dithiothreitol to eliminate blocking IgM HLA-specific antibodies that may mask clinically relevant allosensitization. PMID- 19300183 TI - Adenovirus infection activates akt1 and induces cell proliferation in pancreatic islets1. AB - BACKGROUND: : Accumulated evidence has shown that insulin producing beta-cells in pancreatic islets have the limited potential to regenerate. Adenoviruses have been widely employed to deliver genes of interest into pancreatic islets. This study was aimed at investigating whether adenovirus infection has any impact on the potential of beta-cell proliferation. METHODS: : Human adenovirus type 5 (Ad5) encoding rat insulin promoter driven reporter genes were used to infect freshly isolated pancreatic islets. Western blotting assays were performed to evaluate the expression and activation of key molecules involved in cell survival and proliferation following Ad5 infection. Immunofluorescence staining was employed to identify proliferating cells after culturing the infected and control islets in the presence of BrdU, an analog of thymidine that can be incorporated into the genome of proliferating cells. RESULTS: : Ad5 infection of the islets resulted in expression and activation of Akt1, a key molecule in the PI3 kinase signaling pathway. Accordingly, a higher frequency of islet cell proliferation was detected in Ad5-infected islets than in control islets. DISCUSSION: : These data suggest adenovirus infection can activate beta-cell survival and proliferation machinery, in particular operating through the PI3K/Akt signaling pathway. This information has significant ramification for the use of adenovirus as a gene delivery vehicle for pancreatic islet cells. PMID- 19300184 TI - MRI assessment of ischemic liver after intraportal islet transplantation. AB - BACKGROUND: There is a recent focus on embolization of the portal vein by transplanted islets as a major cause of early graft loss. The resultant ischemia causes necrosis or apoptosis of cells within the liver. Thus, noninvasive assessment of the liver receiving the islet transplant is important to evaluate the status islet grafts. METHODS: This study used noninvasive magnetic resonance imaging (MRI) for assessment of the posttransplant ischemic liver. Syngeneic islets in streprozotocin-induced diabetic mice were used. MRI and morphological liver assessments were performed at 0, 2, and 28 days after transplantation. Histologic assessment of insulin, hypoxia induced factor 1-alpha, and apoptosis were undertaken at similar time points. RESULTS: Ischemic/necrotic regions in the liver were detected by MRI at 2 days but not at 28 days after transplantation and were confirmed histologically. Liver injury was quantified from high intensity areas on T2-weighted images. Insulin release peaked 2 days after transplantation. CONCLUSION: Onset and reversal of liver ischemia due to intraportal islet transplantation are detectable using T2-weighted MRI. These changes coincide with periods of maximum insulin release likely due to partial islet destruction. We propose that MRI, as a noninvasive monitor of graft-related ischemia, may be useful in assessment of liver and islet engraftment after intraportal islet transplantation in a clinical setting. PMID- 19300185 TI - Cost effectiveness of open versus laparoscopic living-donor nephrectomy. AB - BACKGROUND: Kidney transplantation is an essential part of care for patients with end-stage renal disease. The introduction of laparoscopic living-donor nephrectomy (LLDN) has made live donation more advantageous because of less postoperative pain, earlier return to normal activities, and a consequent potential to increase the pool of kidney donors. However, the cost effectiveness of LLDN remains unknown. The aim of this study was to explore the health and cost consequences of replacing open-donor nephrectomy by LLDN. METHODS: Kidney donors were randomized to laparoscopic (n=63) or open surgery (n=59). We obtained data on operating time, personnel costs, length of stay, cost of analgesic, disposable instruments and complications, and indirect costs. Quality of life was captured before the operation and at 1, 6, and 12 months postdonation by means of short form-36. The scores were translated into utilities by means of Brazier's 6D algorithm. RESULTS: The cost per patient was U.S. $55,292 with laparoscopic and U.S. $29,886 with open surgery. The greatest cost difference was in costs attributed to complications (U.S. $33,162 vs. U.S. $4,573). The 1-year quality adjusted life years (QALYs) were 0.780 and 0.765, respectively for laparoscopic and open surgery. This implies a cost of U.S. $1,693,733 per QALY at 12 months follow-up. Sensitivity analyses indicated that the cost of the major complications in the laparoscopic group and magnitude of QALY gain had the greatest impact on cost effectiveness. CONCLUSIONS: The LLDN is an attractive alternative because it, in general, entails less postoperative pain than open surgery, but it is cost effective only with relatively low rates of complications. PMID- 19300186 TI - Impact of a pharmaceutical care program on liver transplant patients' compliance with immunosuppressive medication: a prospective, randomized, controlled trial using electronic monitoring. AB - INTRODUCTION: Compliance with immunosuppressive therapy plays a major role in the long-term success of organ transplantation. Thus, strategies to promote compliance in posttransplant care are of particular interest. At the pharmacy department of the University Hospital Mainz, a program for pharmaceutical care of organ transplant patients has been developed for the first time ever. The main objective of the presented study was to examine the influence of this program on liver transplant patients' compliance with immunosuppressive therapy. METHODS: To measure compliance, medication event monitoring systems were used. Dosing compliance (DC) was calculated for each patient and the mean DC was compared between the two groups. Further direct and indirect methods of measuring compliance served to confirm the electronic compliance data. RESULTS: Pharmaceutical care of liver transplant patients led to a significant increase in compliance with the immunosuppressive therapy. The mean DC of the intervention group was 90%+/-6% compared with 81%+/-12% in the control group (P=0.015). Only two patients (10%) in the intervention group and nine patients (43%) in the control group showed a DC less than 80% (P=0.032). Furthermore, patients in the intervention group were more likely to achieve target blood levels. CONCLUSION: Patients who received pharmaceutical care with traditional patient care showed significantly better compliance with their immunosuppressive medication than patients who received only traditional patient care. Pharmaceutical care proved to be an effective intervention that should be implemented in posttransplant care. PMID- 19300187 TI - Prevalence of urogenital mycoplasmas and ureaplasmas in women after kidney transplantation. AB - BACKGROUND: The prevalence of urogenital mycoplasmas and ureaplasmas in kidney transplant and hemodialyzed patients was studied. METHODS: Vaginal and cervical swabs taken from 40 women of the study group and 40 women of the control group were investigated. Identification of ureaplasmas, Mycoplasma genitalium, and human papillomavirus was performed by polymerase chain reaction. Each vaginal slide was evaluated for bacterial vaginosis. RESULTS: Urogenital mycoplasmas and ureaplasmas were significantly more common in patients from the study group (40%) compared with the control group (27.5%). Mycoplasma hominis and M. genitalium were demonstrated only in a few cases. Ureaplasma parvum was isolated predominantly, but Ureaplasma urealyticum was more common in patients from study group (10%) compared with control group (2.5%). In all U. urealyticum-positive women from the study group, human papillomavirus DNA was detected. CONCLUSION: Our observation showed the necessity of careful examination of possible atypical pathogens in diagnostic materials from hemodialyzed and kidney transplant patients. PMID- 19300188 TI - Prospective monitoring of Epstein-Barr virus DNA in adult renal transplant recipients during the early posttransplant period: role of mycophenolate mofetil. AB - BACKGROUND: Epstein-Barr virus (EBV) is associated with posttransplant lymphoproliferative disease. We monitored the incidence of EBV viraemia in adult renal transplant recipients and investigated the association with clinical parameters. METHODS: Whole blood from 115 renal transplant patients was tested regularly by quantitative polymerase chain reaction (PCR) assay for EBV DNA during the first 90 days posttransplantation. RESULTS: Sixty four of 115 (56%) patients had detectable EBV DNA in blood (>100 copies/mL) on at least one occasion. The median time to first DNA detection was 15 days post-transplant and median viral load was 598 copies/mL (range 119-53,649 copies/mL). Multivariate Cox-regression analyses showed that patients receiving mycophenolate mofetil (MMF) on the day of transplant had a significantly lower risk of EBV viraemia compared to those who received no MMF (Hazard ratio=0.518, 95% CI 0.307-0.875, p=0.014). CONCLUSIONS: EBV viraemia is common during the early posttransplant period in adult renal transplant recipients. Our results suggest a role of MMF in preventing EBV viraemia, however further work is required to identify the mechanism(s) involved. PMID- 19300189 TI - Expression of regulatory T-cell-related molecule genes and clinical outcome in kidney transplant recipients. AB - BACKGROUND: Naturally occurring regulatory T cells have been associated with long term allograft survival. We investigated whether gene transcripts of Treg-related molecules are upregulated or downregulated in kidney transplant recipients with different clinical outcomes and may serve as markers of operative tolerance. METHODS: Expression levels of transcription factor (forkhead box P3 [FOXP3], t bet, and GATA3), regulatory molecule (cytotoxic T-lymphocyte antigen-4, glucocorticoid-induced tumor necrosis factor receptor-related protein, tribbles protein-1, and transforming growth factor-beta), and chemokine receptor (CCR7 and CXCR4) genes were measured in kidney graft recipients with long-term (> or = 9 years) stable renal function (LTS) or chronic rejection (ChrRx). Patients on dialysis and healthy individuals served as controls. RESULTS: The level of FOXP3 transcripts was lower in ChrRx patients than in LTS patients (P<0.01). The highest transforming growth factor-beta transcripts were observed in ChrRx and the highest CCR7 and CXCR4 transcripts were observed in LTS patients. In LTS patients, FOXP3 gene expression was associated with CXCR4 gene expression (P=0.015). FOXP3 and CCR7 transcript levels were higher in LTS patients without calcineurin inhibitor therapy than in LTS patients with calcineurin inhibitors. CONCLUSION: Our results suggest that high expression of FOXP3 and chemokine receptor genes in LTS patients are possible indicators of a regulatory process that contributes to long-term allograft acceptance. Markers that were increased in LTS patients were found to be decreased in ChrRx patients, suggesting that rejection may partly be the result of a lack of this regulatory process. FOXP3 and CCR7 and CXCR4 transcripts might be used as markers to distinguish patients who developed long-term allograft acceptance from patients who are prone to ChrRx. PMID- 19300190 TI - A reevaluation of the role of IgM Non-HLA antibodies in cardiac transplantation. AB - BACKGROUND: Preexisting IgG antibodies to donor human leukocyte antigens (HLA) are a risk factor for rapid allograft rejection. However, non-HLA antibodies, of the IgM class, also called autoreactive antibodies, are not believed to affect graft survival. The aim of this study was to determine the incidence and clinical relevance of pretransplant lymphocytotoxic non-HLA IgM antibodies on long-term cardiac allograft survival. METHODS: A retrospective study of 616 adult recipients of cardiac allografts, transplanted at this center between 1991 and 2003, has been performed. Antibodies in pretransplant sera were initially defined using complement-dependent cytotoxicity assays, and subsequently analyzed for HLA specificities using solid phase assays. RESULTS: HLA antibodies were present in 69 of 616 heart recipients (58 IgG, 11 IgM); in 22 of these, the antibodies were donor-specific. Non-HLA IgM antibodies were detected in 59 of 616 recipients who did not have HLA-specific antibodies; these patients had a 1, 2, 5, and 10 year survival of 55.9%, 54.2%, 49.9%, and 43.3% compared with 75.8%, 73.7%, 66.6%, and 52.8% for those without antibodies (P=0.0085 log-rank test). Multivariate analysis demonstrated pretransplant non-HLA IgM antibodies to be an independent risk factor for mortality (P=0.0001). Myocardial histology of postmortem heart and cardiac biopsies suggested an association with ischemic damage and "primary" allograft failure. CONCLUSIONS: We propose the hypothesis that the presence of cytotoxic IgM antibodies to non-HLAs before heart transplantation maybe a risk factor for early allograft failure. PMID- 19300191 TI - Predictive factors of outcome in patients transplanted for hepatitis B. AB - BACKGROUND: This study aimed to (1) identify the variables that affect graft and patient survival in recipients transplanted for hepatitis B virus (HBV) disease; and (2) assess factors associated with an increased risk of graft cirrhosis at 5 years after liver transplantation (LT). METHODS AND RESULTS: A total of 104 chronically infected HBV patients were considered for this study and all received tacrolimus- or cyclosporine A (CSA)-based immunosuppressive regimens. The overall 5-year patient and graft survival rates were 80% and 73%, respectively. Univariate Cox proportional hazards regression analysis indicated that older recipient age and higher body mass index (BMI) at LT, LT more than or equal to 1998, arterial hypertension, hyperlipidemia, and CSA-based immunosuppression correlated with decreased patient survival. In the multivariate model, advanced recipient age, higher BMI, CSA-based immunosuppressive therapy, and increasing cold ischemia time were associated with worse patient survival. Recipient age and BMI at time of LT and posttransplant hypertriglyceridemia also affected graft survival. Log-rank analysis showed that a viral load of more than 10 copies/mL and antiviral therapy at LT, post-LT biliary complications, HBV recurrence, nucleos(t)ide analogue monoprophylaxis (without hepatitis B immunoglobulin), and short-term (< or = 1 year) mycophenolate mofetil therapy were significant risk factors for graft cirrhosis within 5 years of LT. CONCLUSION: Various recipient factors at the time of LT and post-LT virological status, antiviral prophylaxis, cholestasis, cardiovascular risk profile, and immunosuppressive regimen affect the outcome of HBV patients after LT. Prospective studies are warranted to define optimal immunosuppression for recipients transplanted for hepatitis B. PMID- 19300192 TI - Rises and falls in donor-specific and third-party HLA antibody levels after antibody incompatible transplantation. AB - BACKGROUND: After human leukocyte antigen (HLA) antibody-incompatible transplantation, donor specific and third party HLA antibodies may be found, and their levels fall in a donor-specific manner during the first month. However, these changes have not been previously described in detail. METHODS: Donor specific HLA antibody (DSA) and third-party HLA antibody (TPA) levels were measured using the microbead method in 44 presensitized patients who had renal transplantation. RESULTS: DSA+TPA fell in the first 4 days after transplantation, and greater falls in DSA indicated absorption by the graft. This occurred for class I (57.8% fall compared with 20.2% for TPA, P<0.0005), HLA DR (63.0% vs. 24.3%, P<0.0004), and for HLA DP/DQ/DRB3-4 (34% vs. 17.5%, P=0.014). Peak DSA levels occurred at a mean of 13 days posttransplant, and they were higher than pretreatment in 25 (57%) patients and lower in 19 (43%) patients (P=ns). The risk of rejection was associated with peak DSA levels; 15 of 25 (60%) patients with DSA at median fluorescence intensity (MFI) more than 7000U experienced rejection, compared with 4 of 7 (57%) patients with peak DSA MFI 2000 to 7000U, and 2 of 12 (17%) patients with peak DSA MFI less than 2000U (P<0.02). DSA levels subsequently fell in a donor specific manner compared to TPA. CONCLUSION: DSA levels may change markedly in the first month after antibody incompatible transplantation, and the risk of rejection was associated with higher pretreatment and peak levels. PMID- 19300193 TI - Outcome of renal transplant recipients admitted to an intensive care unit: a 10 year cohort study. AB - BACKGROUND: Epidemiology and prognosis of severe complications related to renal transplantation requiring admission to intensive care unit (ICU) have not been assessed precisely. This study was undertaken to evaluate the outcome in this population and to identify the factors of prognosis. METHODS: All records of adult renal transplant recipients admitted to our ICU from 1997 to 2007 were reviewed including transplant variables, clinical and biological parameters, use of mechanical ventilation, catecholamine support, or dialysis or both. Mortality was assessed and data were analyzed to identify predictive factors of outcome. RESULTS: Twenty-seven women and 30 men, median age 54 years, were included in the study. Eighteen patients were oliguric, 35 were mechanically ventilated, 32 underwent hemodialysis, and 36 needed catecholamine. Twenty-three patients died (40.3%), a mortality significantly higher than in a matched by age and gravity scores control group of nontransplant ICU patients. By univariate analysis, survivors had a significantly lower ICU severity scores, a higher mean arterial pressure, a higher Glasgow Coma Score, a higher serum albumin, and a lower serum lactate on ICU admission. The need for catecholamine support, mechanical ventilation or dialysis or both during the ICU stay worsens the outcome significantly. Using the multivariate analysis, only the mean arterial pressure and the need for mechanical ventilation were predictive of mortality. CONCLUSION: The incidence of severe transplant-related complications requiring an admission to an ICU was at 16 of 1000 patients year with a mortality rate higher than the general ICU population (40% vs. 20%). These data suggest that immunosuppressive treatment of transplant patients with severe complications worsens significantly their outcome. PMID- 19300194 TI - Donor-derived mesenchymal stem cells suppress alloreactivity of kidney transplant patients. AB - BACKGROUND: Human mesenchymal stem cells (MSC) have immunosuppressive capacities. Although their efficacy is currently studied in graft-versus-host disease, their effect on alloreactivity in solid organ transplant patients is unknown. In this study, the immunosuppressive effect of MSC on recipient anti-donor reactivity was examined before and after clinical kidney transplantation. METHODS: Anti-donor reactivity was established in pretransplant and posttransplant mixed lymphocyte reactions (MLR) of 14 living-kidney donor-recipient pairs. MSC from donors and third-party controls were added to the MLR in a ratio of 1:5. RESULTS: MSC were isolated from donor perirenal fat and showed multilineage differentiation potential and the capacity to inhibit lymphocyte proliferation. The immunosuppressive effect of MSC was dose dependent and mediated by cell-membrane contact and soluble factors, including interleukin-10 and indoleamine 2,3 dioxygenase.Donor-derived MSC significantly inhibited the recipient anti-donor reactivity before and 1 month after transplantation. This effect was independent of human leukocyte antigen background of MSC. Flow cytometric analysis showed that MSC inhibited the proliferation of CD4+ and CD8+ T-lymphocyte subsets in pretransplant and posttransplant donor-directed MLR, whereas MSC had no effect on B- or natural killer-cell proliferation. CONCLUSION: Donor MSC significantly inhibited the proliferation of alloactivated recipient T cells before and after kidney transplantation. We believe these findings should encourage MSC-based intervention in clinical organ transplantation. PMID- 19300195 TI - T-cell reactivity during tapering of immunosuppression to low-dose monotherapy prednisolone in HLA-identical living-related renal transplant recipients. AB - BACKGROUND: In many transplant centers, human leukocyte antigen (HLA)-identical living-related (LR) renal transplant recipients receive standard maintenance immunosuppression from 1 year after transplantation. We questioned whether discontinuation of azathioprine (AZA) or mycophenolate mofetil (MMF) influenced T cell reactivity, circulating dendritic cell (DC) subsets numbers and their maturation status. METHODS: Twenty-nine HLA-identical LR renal transplant recipients were withdrawn from AZA or MMF. Thereafter, the patients received only prednisolone. T-cell reactivity was determined by interferon-gamma (n=23), interleukin (IL)-10 (n=16), and granzyme B (n=10) Elispot assays. Circulating DC subset numbers and their maturation status determined by CCR2, CCR5, CCR7, and CD83 expression were measured by flow cytometry (n=12). RESULTS: The number of donor, third-party, and tetanus toxoid-reactive interferon-gamma and granzyme-B producing cells was not affected after withdrawal of immunosuppression. Discontinuation of AZA or MMF resulted in significant increased numbers of third party (P=0.003) and tetanus toxoid-reactive (P=0.008) IL-10 producing cells, and a trend in higher numbers of donor-reactive IL-10 producing cells (P=0.06). No effect was found on the number of circulating DC subsets, but DC was shifted toward a more mature phenotype. CONCLUSIONS: In HLA-identical LR renal transplant recipients, therapy with AZA and MMF suppress the IL-10 production and the maturation of DC. This suggests that these immunosuppressants may hinder suppression of immune responses in general, including allogeneic responses. PMID- 19300196 TI - Conjunctival epithelial cells with donor-derived genome after human haematopoietic stem-cell transplantation. AB - To detect conjunctival epithelial cells with donor derived genome following human haematopoietic stem cell transplantation. Conjunctival biopsies from allotransplanted females were stained for the Y-chromosome, cytokeratin, and CD45 followed by a DAPI counterstain and CD68 staining and were then evaluated by laser-scanning confocal microscopy and 3D analysis. In 5 patients donor derived epithelial cells identified as Y+/DAPI+/CK+/CD45-/CD68-/epithelial morphology could be found (57%) with a mean incidence of 5.4% (range 2.42% - 7.94%). Our study demonstrates that besides the already published results in liver, colon, lung, skin and oral mucosa, epithelial cells with donor-derived genome also emerge in the conjunctiva following human HSCT. The biological significance and the underlying mechanism of these findings need further clarification. PMID- 19300197 TI - Expanding postmortem donor pool using steatotic liver grafts: a new look. AB - BACKGROUND: Clinical demand for liver transplant steadily grows while organs offer has reached a plateau years ago. To expand the donor liver pool, various options have been considered including acceptance of suboptimal donors and steatotic grafts, with a risk of poorer outcomes. The latter risk and its relation to the grade of liver graft steatosis have been studied in this prospective clinical study. METHODS: One hundred eighteen consecutive liver transplantation (115 patients) performed between May 2002 and March 2008 were prospectively analyzed. According to the grade of steatosis on a 2 hr postreperfusion biopsy, four groups were considered: absence (<5%) (n=34), mild (<30%) (n=40), moderate (30%-60%) (n=23), or severe steatosis (> or = 60%) (n=21). Donors and recipients demographic data, and patients and grafts survival rates were compared among the four groups. RESULTS: Eighty-four (71%) grafts presented some degree of steatosis (macrosteatosis: 19.5%, microsteatosis: 47%, mix type: 33.5%). Patient and graft survival were significant lower in the "severe steatosis" group, as a whole. Grafts with less than 30% predominant macro , or microsteatosis also had poorer outcomes with lower patient and graft survival rates. CONCLUSION: Steatotic liver grafts were used on a large scale (71%) in this clinical series. The analysis confirms that using grafts with moderate (>30%) and severe steatosis (>60%) have a negative impact on outcomes. The authors conclude that using these grafts allow a significant increase in organ offer that counterbalances the negative outcome for patients who are not offered a transplant, and this supports the need for further clinical research. PMID- 19300198 TI - Assessment of belatacept-mediated costimulation blockade through evaluation of CD80/86-receptor saturation. AB - BACKGROUND: The selective inhibitor of T-cell costimulation, belatacept, blocks CD28-mediated T-cell activation by binding CD80 and CD86 on antigen-presenting cells. Understanding the extent to which belatacept binds to its targets in patients may enable correlation of belatacept exposure to receptor saturation as a pharmacodynamic measure of costimulation blockade. METHODS: Flow cytometry based receptor competition assays were developed to monitor concentration dependent occupancy of CD80 and CD86 receptors in whole blood and dendritic cell cultures in vitro. Receptor occupancy was correlated with inhibition of mixed leukocyte reactions and clinical validation was obtained by comparing receptor saturation in whole blood from healthy volunteers and in de novo renal transplant recipients participating in studies comparing cyclosporine and belatacept-based immunosuppression. RESULTS: Belatacept saturated CD80 and CD86 receptors in whole blood and dendritic cell cultures, although the belatacept concentrations required for CD86-receptor saturation were approximately 10-fold higher than those required for CD80 saturation (IC50=0.102 microg/mL vs. 0.009 microg/mL). Primary alloresponses were inhibited at the belatacept concentration required for CD86-receptor saturation, but not at the lower concentration needed to saturate CD80. Whole blood from belatacept-treated patients had significantly lower levels of free CD86 receptors versus pretransplant levels, healthy volunteers, or cyclosporine-treated patients. CD86-receptor saturation correlated with belatacept dose/dose frequency and remained consistently more than 80%. CONCLUSIONS: These results suggest that belatacept-mediated inhibition of alloresponses involved in transplant rejection correlates with CD86 saturation, indicating that CD86-receptor occupancy may be a valid pharmacodynamic measure of costimulation blockade and provide the first direct clinical evidence that belatacept binds to one of its targets. PMID- 19300199 TI - Prospective, randomized trial on late steroid withdrawal in pediatric renal transplant recipients under cyclosporine microemulsion and mycophenolate mofetil. AB - BACKGROUND: : Many transplant centers practice late steroid withdrawal after pediatric renal transplantation, but evidence-based data on the overall risk-to benefit ratio in this patient population are lacking. METHODS: : We therefore conducted the first prospective, randomized, open-label multicenter study to validate this strategy: 42 low-immunologic risk pediatric kidney allograft recipients, aged 10.3+/-4.3 years, on cyclosporine microemulsion, mycophenolate mofetil, and corticosteroids were randomly assigned, more than or equal to 1-year posttransplant, to continue steroids or to withdraw over 3 months. This report contains the 1-year results. RESULTS: : In response to steroid withdrawal, patients experienced a significant catch-up growth with a mean standardized height gain of 0.3+/-0.1 standard deviation score (SDS) per year (P<0.05 vs. control), whereas mean height SDS in the control group did not change (0.0+/-0.1 SDS). Standardized body mass index declined significantly by 0.68+/-0.23 SDS after steroid withdrawal, but rose significantly by 0.26+/-0.34 SDS in the control group. Patients off steroids had less frequent arterial hypertension (50% vs. 87.5% (P<0.05) and significantly lower serum cholesterol (by 21%) and triglyceride values (by 36%) than control patients. Patient and graft survival were 100%. The incidence of acute rejection episodes in the steroid-withdrawal group was 1 of 23 (4%) compared with 1 of 19 (5%) in controls. Transplant function remained stable in both groups. CONCLUSION: : Late steroid withdrawal in low-immunologic risk European pediatric kidney transplant recipients on cyclosporine microemulsion and mycophenolate mofetil is not associated with an increased rate of acute rejection episodes, enables catch-up growth and ameliorates cardiovascular risk factors. PMID- 19300200 TI - VKORC1 haplotypes and long-term renal allograft survival. PMID- 19300201 TI - Norwegian scabies associated with herpes simplex infection in a renal transplant patient. PMID- 19300203 TI - Intraductal papillary mucinous neoplasms of the pancreas: differentiation of malignant and benign tumors by endoscopic ultrasound findings of mural nodules. AB - BACKGROUND AND AIM: Intraductal papillary mucinous neoplasms (IPMNs)have a wide pathologic spectrum and it is difficult to differentiate malignant from benign tumors. The aim of this study was to identify predictors of malignancy using contrast-enhanced endoscopic ultrasound (CE-EUS). SUBJECTS AND METHODS: In our institute, main duct type and mixed type IPMNs, branch duct type IPMNs with mural nodules, and IPMNs with coexistent invasive ductal cancer were indications for surgery. Eighty-seven IPMNs (14 main duct, 25 mixed, and 48 branch duct type) were resected and CE-EUS findings were compared with pathologic findings. Twelve clinicopathological variables and CE-EUS morphologic findings were assessed.Mural nodules defined as blood flow supplied protrusions were classified into 4 types: type I: low papillary nodule, type II: polypoid nodule, type III:papillary nodule, and type IV: invasive nodule. RESULTS: Forty-two, 26, 16, and 3 were pathologically diagnosed as adenoma,noninvasive carcinoma, invasive IPMNs, and coexistent invasive ductal cancer, respectively. Multivariable logistic regression analysis showed that types III/IV mural nodule (odds ratio = 10.8; 95% confidential intervals = 2.75-56.1) and symptomatic IPMNs (odds ratio = 4.31; 95% confidential intervals = 1.37-14.7) were significant for malignancy. For mural nodule diameter, invasive IPMNs were significantly larger, but types III and IV mural nodules were more frequently associated with malignancy, particularly invasive cancer, at 88.9% and 91.7%, respectively. The diagnosis of IPMNs with types III or IV mural nodule as malignant resulted in a sensitivity of 60%, specificity of 92.9%, and accuracy of 75.9%. CONCLUSIONS: In conclusion, new morphologic criteria were useful to identify the malignant potentials of IPMNs. PMID- 19300202 TI - Ten-year incidence and risk factors of bone fractures in a cohort of treated HIV1 infected adults. AB - In the ANRS CO8 APROCO-COPILOTE cohort of patients treated with combination antiretroviral therapy since 1997-1999, the incidence density of bone fractures was 3.3 for 1000 patient-years [95% confidence interval (CI) = 2.0-4.6]. The rate was 2.9-fold (95% CI = 1.3-6.5) higher among patients with excessive alcohol consumption and 3.6-fold (95% CI = 1.6-8.1) higher in those with hepatitis C virus (HCV) coinfection. Specific monitoring of HCV/HIV-coinfected patients and active promotion of alcohol cessation should be recommended for the prevention of bone fractures. PMID- 19300205 TI - When should prophylactic antibiotics be given? PMID- 19300206 TI - Recombinant activated factor VII in abdominal aortic emergencies. PMID- 19300209 TI - In favor of the Jarnagin-Blumgart classification. PMID- 19300211 TI - Laparoscopic colectomy survival benefit for colon cancer: is evidence from a randomized trial true? PMID- 19300215 TI - Physician-specific OPPE models in academic medical centers. PMID- 19300217 TI - Effect of subjective preoperative variables on risk-adjusted assessment of hospital morbidity and mortality. AB - OBJECTIVE: To examine the influence of American Society of Anesthesiologists Physical Status Classification (ASA) and preoperative Functional Health Status (FHS) variables on risk-adjusted estimates of surgical quality and to assess whether classifications are inflated at some hospitals. BACKGROUND: ASA and FHS are influential in risk-adjusted comparisons of surgical quality. However, because ASA and FHS are subjective they can be inflated, making patients appear more ill than they actually are, and crediting hospitals for a sicker patient population. METHODS: We identified 28,751 colorectal surgery patients at 170 hospitals participating in the American College of Surgeon's National Surgical Quality Improvement Program (ACS NSQIP) during 2006 to 2007. Logistic regression models were developed for morbidity and mortality with and without inclusion of ASA and FHS. Hospital quality rankings from the different models were compared. RESULTS: Morbidity and mortality rates were 24.3% and 3.9%, respectively. Percents of patients in ASA classes I through V were 3.3%, 46.4%, 41.5%,8.3%, and 0.7% and that were independent or partially or totally dependent were 89.2%, 7.2%, and 3.6%, respectively. Models that included ASA and FHS exhibited slightly better fit (Hosmer-Lemshow statistic) and discrimination(c-statistic) than models without both these variables, though magnitudes of differences were consistent with chance. There was inconsistent evidence for improper assignment of ASA and FHS. CONCLUSIONS: The small improvements in model quality when both ASA and FHS are present versus absent, suggest that they make a unique contribution to assessing severity of preoperative risk. With little indication that these subjective variables are subject to an important level of institutional bias, it is appropriate that they be used to assess risk-adjusted surgical quality. Periodic monitoring for inappropriate inflation of ASA status is warranted. PMID- 19300218 TI - M.A.I.D.: a prognostic score estimating probability of healing in chronic lower extremity wounds. AB - OBJECTIVE: To evaluate a wound-based prognostic score for chronic lower extremity wounds suitable for daily routine use capable of predicting long-term healing. SUMMARY BACKGROUND DATA: The main obstacle in the treatment of chronic wounds is to estimate long-term clinical outcome. For diabetic foot ulcers, several ulcer, and nonulcer-related risk factors associated with impaired healing have been described in the past. METHODS: A new chronic lower extremity ulcer score (M.A.I.D.) was created out of 4 clinically defined parameters, namely palpable pedal pulses (I), wound area (A), ulcer duration (D), and presence of multiple ulcerations (M). Palpable pedal pulses were categorized by the absence (scored as 1) or presence (scored as 0) of pedal pulses, while wounds >4 cm were scored as 1 and wounds < or = 4 cm as 0. Ulcers lasting more than 130 days were categorized as 1 and wounds with a duration of <130 days as 0. Patients with multiple ulcerations were graded as 1 (=1) compared with those with single ulcers (=0). M.A.I.D. was calculated by adding these separate scores to a theoretical maximum of 4. RESULTS: Two thousand nineteen consecutive patients with 4004 wounds were included. When patients were divided into subgroups with the same M.A.I.D., we showed a decreasing probability of healing for ulcers with higher M.A.I.D. scores. An increase in the M.A.I.D. by 1 score-point reduced the chance for healing by 37%. Similarly, the higher the ulcer score, the larger the initial wound area, the longer the wound history, and the more likely the occurrence of soft-tissue infection during follow-up. CONCLUSIONS: This new chronic lower extremity ulcer score is capable of anticipating long-term probability of healing by combining 4 clinically assessable parameters. However, adequate and standardized wound care is an indispensable prerequisite for M.A.I.D. to be a valid diagnostic tool in daily clinical routine. PMID- 19300219 TI - Emergency femoral hernia repair: a study based on a national register. AB - OBJECTIVE: To describe the characteristics of femoral hernias and outcome of femoral repairs, with special emphasis on emergency operations. BACKGROUND: Femoral hernias account for 2% to 4% of all groin hernias. However, the lack of large-scale studies has made it impossible to draw conclusions regarding the best management of these hernias. METHODS: The study is based on patients 15 years or older who underwent groin hernia repair 1992 to 2006 at units participating in the Swedish Hernia Register. RESULTS: Three thousand nine hundred eighty femoral hernia repairs were registered, 1490 on men and 2490 on women: 1430 (35.9%) patients underwent emergency surgery compared with 4.9% of the 138,309 patients with inguinal hernias. Bowel resection was performed in 22.7% (325) of emergent femoral repairs and 5.4% (363) of emergent inguinal repairs. Women had a substantial over risk for undergoing emergency femoral surgery compared with men (40.6% vs. 28.1%). An emergency femoral hernia operation was associated with a 10 fold increased mortality risk, whereas the risk for an elective repair did not exceed that of the general population. In elective femoral hernias, laparoscopic (hazard ratio, 0.31; 95% confidence interval, 0.15-0.67) and open preperitoneal mesh (hazard ratio, 0.28; confidence interval, 0.12-0.65) techniques resulted in fewer re-operations than suture repairs. CONCLUSIONS: Femoral hernias are more common in women and lead to a substantial over risk for an emergency operation, and consequently, a higher rate of bowel resection and mortality. Femoral hernias should be operated with high priority to avoid incarceration and be repaired with a mesh. PMID- 19300220 TI - Surgical aspects in the treatment of patients with unilateral wilms tumor: a report from the SIOP 93-01/German Society of Pediatric Oncology and Hematology. AB - OBJECTIVE: To assess surgical aspects in the treatment of children with unilateral Wilms tumor based on data from the Cooperative Tumor Study SIOP 93-01 of the German Society of Pediatric Hematology and Oncology. SUMMARY BACKGROUND DATA: Although multiple international study trials exist for the treatment of nephroblastoma, the impact of surgical details and the outcome of the patients have not yet been described comparing different approaches of the trials. METHODS: Treatment results of SIOP 93-01 of the German Society of Pediatric Hematology and Oncology were analyzed regarding frequencies of operations by surgeons and hospitals, surgical approaches, and operating subspecialties. Special attention was given to surgical complications, postoperative tumor stages and event-free survival. RESULTS: Data sets from 757 of 1020 registered patients were received for evaluation. A unilateral Wilms tumor was observed in 512 of 757 children.Median follow-up was 4.8 years (1.2-10.7). Event-free survival rates were comparable for frequencies of operation by surgeons and hospitals, surgical approaches, and surgical specialties. Intraoperative tumor rupture rates were 12% in primarily operated patients (protocol violations) versus 3.2% inpatients after preoperative chemotherapy. There were 7% intraoperative ruptures for hospitals and surgeons performing 1 operation per year, and 3%when more than 4 operations per year were carried out. Sampling of hilar lymph nodes was often incomplete for all surgical subspecialties. CONCLUSION: While the event-free survival for all groups is equal, there may be some long-term complications as a result of the more intensified therapy required for patients who suffer intraoperative ruptures. This will be defined only with longer term studies of late effects of the more intensified therapy. There is,however an increased rate of complications and ruptures associated with the use of midline laparotomy rather than a transverse or thoracoabdominal incision. PMID- 19300221 TI - The epidemiology of Candida colonization and invasive candidiasis in a surgical intensive care unit where fluconazole prophylaxis is utilized: follow-up to a randomized clinical trial. AB - OBJECTIVE: To determine whether Candida glabrata colonization and invasive candidiasis (IC) increased among critically ill surgical patients 3 years after the introduction of fluconazole prophylaxis to a surgical intensive care unit (SICU). SUMMARY BACKGROUND DATA: Fluconazole prophylaxis has been shown in randomized clinical trials to reduce the occurrence of candidiasis in some patient populations, including high-risk SICU patients. One such trial was performed in The Johns Hopkins Hospital SICU in 1998. Whether the epidemiology of Candida colonization and IC has changed in SICUs where fluconazole prophylaxis is routinely utilized has not been adequately studied. METHODS: We conducted a prospective, observational study of subjects admitted for > or = 3 days to the SICU of a large, urban, academic medical center, where fluconazole prophylaxis had been utilized for approximately 3 years. Surveillance fungal cultures of rectal/fecal swabs, urine, and endotracheal aspirates were performed on admission to the SICU, once weekly, and upon discharge from the SICU. Demographic and clinical data were collected. C. glabrata colonization and IC prevalence among patients in the prospective cohort were compared with the prevalence among SICU patients enrolled in the 1998 clinical trial of fluconazole for the prevention of candidiasis that was performed at the same institution. RESULTS: C. glabrata colonization was not significantly more common among patients in the 2003 cohort as compared with patients in the 1998 trial (adjusted odds ratio [OR]: 0.90, 95% confidence interval [CI]: 0.57-1.41). Patients with IC in the 2003 cohort were not more likely than those in the 1998 trial to have IC due to C. glabrata (adjusted OR: 1.93, 95% CI: 0.20-18.98), while patients with IC in the 2003 cohort were less likely than patients in the 1998 trial to have acquired IC in the ICU (adjusted OR: 0.08, 95% CI: 0.009-0.82). CONCLUSIONS: There was no increase in C. glabrata colonization or in the proportion of IC due to C. glabrata after a 3-year period of routine fluconazole prophylaxis for selected SICU patients. PMID- 19300222 TI - Selective nonoperative management of liver gunshot injuries. AB - OBJECTIVE: Nonoperative management (NOM) of liver gunshot injuries is yet to gain general acceptance. The aim of this study was to assess the feasibility and safety of selective NOM of liver gunshot injuries. PATIENTS AND METHODS: A prospective, protocol-driven study, which included all liver gunshot injuries admitted to a level I trauma center, was conducted over a 4-year period. Patients with right-sided thoracoabdominal, and right upper quadrant gunshot wounds with or without localized right upper quadrant tenderness underwent contrasted abdominal computed tomography scan evaluation to detect the presence of a liver injury. Patients with confirmed liver injuries were observed with serial clinical examinations. Outcome parameters included need for delayed laparotomy, complications, length of hospital stay, and survival. RESULTS: During the study period, 63 patients with liver gunshot injuries were selected for NOM. The mean injury severity score was 19.6 (range, 4-34). Simple liver injuries (grades I and II) occurred in 26 (41.3%) patients and complex liver injuries (grades III, IV, and V) occurred in 37 (58.7%) patients. Associated injuries included 14 (22.2%) kidney, 44 (69.8%) diaphragm, 43 (68.3%) lung contusion, 42 (66.7%) hemothorax and/or pneuomothorax, and 21 (33.3%) rib fractures. Five patients required delayed laparotomy resulting in successful NOM rate of 92%. Complications included liver abscess (3), biliary fistula (3), retained hemothorax (4), and nosocomial pneumonia (5). The mean hospital stay was 6.1 (range, 3-23 days). There was no mortality. CONCLUSION: The NOM of appropriately selected patients with liver gunshot injuries is feasible, safe, and effective, regardless of the liver injury severity. PMID- 19300223 TI - The motor fibers of the recurrent laryngeal nerve are located in the anterior extralaryngeal branch. AB - OBJECTIVE: This study aimed to establish the prevalence of extralaryngeal bifurcation of the recurrent laryngeal nerve (RLN) and investigate the location of the motor fibers to the intrinsic muscles of the larynx within the branches of the RLN. SUMMARY OF BACKGROUND DATA: Recognition of extralaryngeal branching of the RLN is important, because inadvertent division of a branch may lead to significant vocal cord palsy despite the operator believing the nerve was preserved. METHODS: Prospective operative data on branching of the RLN were collected in 579 patients undergoing thyroidectomy or open parathyroidectomy over a 3 year period and nerve integrity monitoring was utilized to document the position of the motor fibers of the last 176 RLNs. Adduction of the vocal cords was detected by the electromyography-endotracheal tube and abduction by finger palpation of muscle contraction in the posterior cricoarytenoid. RESULTS: A total of 838 RLN were studied (right--432, left--406). Bifurcations occurred on the right in 111 (25.7%) and left 93 (22.9%). Bilateral bifurcation occurred in 23 (8.9%) of bilaterally dissected nerves. Overall 176 RLNs in 118 patients were assessed by the nerve integrity monitoring. Of these 41 (23.3%) were bifid RLN. In all 41 (100%) cases of bifid RLN, motor fibers for both adduction and abduction of the vocal cords were located exclusively in the anterior branches of RLN, and none in the posterior branches. CONCLUSION: Extralaryngeal bifurcation of RLN is a common anatomical variant. The motor fibers of RLN are located in the anterior branch, for both adduction and abduction. Great care is therefore required following the presumed identification of the RLN to ensure there is no unidentified anterior branch. PMID- 19300224 TI - Interobserver variation in the histopathologic reporting of key prognostic parameters, particularly clark level, affects pathologic staging of primary cutaneous melanoma. AB - BACKGROUND: Accurate reporting of prognostically important histopathologic parameters is critical for optimal management of melanoma patients. Previous studies have shown that assessment of Breslow thickness and ulceration is more reproducible between pathologists than estimation of Clark level. However, detailed quantification of the magnitude of variation between observers and its effect on melanoma staging has not been reported. METHODS: Nine hundred twelve melanoma patients referred to the Sydney Melanoma Unit (SMU) with pathology reports from referring pathology services and reports following histopathologic review by SMU pathologists were studied. Breslow thickness, Clark level, ulceration, and pathologic staging (American Joint Committee on Cancer, 2002) data were extracted from each matched pair of SMU and non-SMU reports. The degree of concordance for these parameters was assessed. RESULTS: There was excellent agreement between SMU and non-SMU pathologists in the assessment of Breslow thickness (kappa = 0.883; intraclass correlation coefficient = 0.968) and ulceration (kappa = 0.832), while that for Clark level was only moderate (kappa = 0.627). The degree of concordance in the assessment of overall pathologic stage was excellent (96.2%), but there was poor agreement for the evaluation of substages IB (77.3%) and IIB (72.4%), due largely to poor Clark level concordance. CONCLUSIONS: Although there was a high level of agreement between pathologists in the assessment of overall pathologic stage, the significant level of discordance in pathologic substaging suggests that accurate and reliable substage-based prognostic prediction may not be possible in all cases. It will be important to consider these issues in future revisions of the American Joint Committee on Cancer melanoma staging system. PMID- 19300225 TI - Surgeon volume impacts hospital mortality for pancreatic resection. AB - OBJECTIVE: Improved outcomes after pancreatic resection (PR) by high volume (HV) surgeons have been reported in single center studies, which may be confounded with potential selection and referral bias. We attempted to determine if improved outcomes by HV surgeons are reproducible when patient demographic factors are controlled at the population level. METHODS: Using the Nationwide Inpatient Sample, discharge records with surgeon identifiers for all nontrauma PR (n = 3581) were examined from 1998 to 2005. Surgeons were divided into 2 groups: (HV; > or = 5 operations/year) or low volume (LV; <5 operations/year). We created a logistic regression model to examine the relationship between surgeon type and operative mortality while accounting for patient and hospital factors. To further eliminate differences in cohorts and determine the true effect of surgeon volume on mortality, case-control groups based on patient demographics were created using propensity scores. RESULTS: One hundred thirty-four HV and 1450 LV surgeons performed 3581 PR in 742 hospitals across 12 states that reported surgeon identifier information over the 8-year period. Patients who underwent PR by HV surgeons were more likely to be male, white raced, and a resident of a high income zip code (P < 0.05). Significant independent factors for in-hospital mortality after PR included increasing age, male gender, Medicaid insurance, and surgery by HV surgeon. HV surgeons had a lower adjusted mortality compared with LV surgeons (2.4% vs. 6.4%; P < 0.0001). CONCLUSIONS: After controlling for patient demographics and factors, pancreatic resection by a HV surgeon in this case-controlled cohort was independently associated with a 51% reduction in in hospital mortality. PMID- 19300226 TI - Liver resection with selective hepatic vascular exclusion: a cohort study. AB - OBJECTIVE: To report our experience on the safety and efficacy of hepatic resection under selective hepatic vascular exclusion (SHVE). METHODS: SHVE was used in 246 consecutive patients undergoing major or complex liver resection in our center. Preoperative demographic and clinical data, details of the surgical procedure, pathologic diagnosis, postoperative course, and complications were collected prospectively. RESULTS: From January 2000 to July 2007, liver resections were performed under SHVE in 246 patients; total SHVE, right partial SHVE, and left partial SHVE in 145, 54, and 47 patients, respectively. SHVE was converted to total hepatic vascular exclusion in 3 patients because the tumor invaded the wall of the inferior vena cava. Hemodynamic tolerance to SHVE was excellent, with only a slight increase in systemic and pulmonary vascular resistance during clamping. There were no deaths and the morbidity rate was 24.8%. The mean hospital stay was 9.6 days (range, 8-18). CONCLUSION: Our study showed that SHVE was safe, efficacious, and it was applicable to liver tumors which were near, but had not invaded into the inferior vena cava. PMID- 19300227 TI - Operative blood loss independently predicts recurrence and survival after resection of hepatocellular carcinoma. AB - OBJECTIVE: To determine if the degree of blood loss during resection of hepatocellular carcinoma (HCC) is predictive of recurrence and long-term survival. BACKGROUND: Several studies have addressed the impact of blood transfusion on survival and recurrence after liver resection for HCC. However, the independent effect of intraoperative estimated blood loss (EBL) on oncologic outcome is unclear. METHODS: From our prospective database, we identified 192 patients who had a partial hepatectomy for HCC from 1985 to 2002. Clinicopathologic predictors of EBL were identified using logistic regression. Overall survival (OS), disease-specific survival (DSS), and recurrence free survival (RFS) were assessed using the Kaplan-Meier and Cox regression methods. RESULTS: The median patient age was 64 (range, 19-86) and 66% were men. All patients had histologically proven HCC. The median follow-up time was 34 months (range, 1-297). Factors associated with increased EBL on multivariate analysis were male gender, vascular invasion, extent of hepatectomy, and operative time (P < 0.01). EBL and vascular invasion were independent predictors of OS and DSS. Only EBL was significantly associated with RFS on multivariate analysis (P = 0.02). Additionally, we found a significant inverse correlation between increasing levels of EBL and length of DSS (P = 0.01). CONCLUSIONS: The magnitude of EBL during HCC resection is related to biologic characteristics of the tumor as well as the extent of surgery. Increased intraoperative blood loss during HCC resection is an independent prognostic factor for tumor recurrence and death. PMID- 19300228 TI - Sequential preoperative ipsilateral hepatic vein embolization after portal vein embolization to induce further liver regeneration in patients with hepatobiliary malignancy. AB - OBJECTIVE: To assess the effect of ipsilateral hepatic vein embolization (HVE) performed after portal vein embolization (PVE) on liver regeneration. SUMMARY BACKGROUND DATA: PVE induces shrinkage of the embolized lobe and compensatory enlargement of the non-embolized lobe, but it does not always induce sufficient liver regeneration. There was no effective method to accelerate liver regeneration in addition to PVE yet. METHODS: During a 1-year study period, preoperative HVE were performed on 12 patients who had shown limited liver regeneration after PVE awaiting right hepatectomy. The right hepatic vein was embolized with multiple coils after insertion of vena cava filters or vascular plugs. RESULTS: No HVE procedure-related complications occurred, but embolization of the wrong hepatic vein trunk occurred in 1 patient. The increase in blood liver enzymes after HVE was comparable with that after PVE alone. In 9 patients who underwent hepatectomy, the proportions of future liver remnant volume to total liver volume were 34.8% +/- 1.5% before PVE, 39.7% +/- 0.6% 1 to 2 weeks after PVE, 44.2% +/- 1.1% 2 weeks after HVE, and 64.5% +/- 6.2% 1 week after right hepatectomy. Cirrhotic livers showed lower regeneration rates following HVE after PVE and 1 patient underwent hepatectomy 17 months after HVE. Immunohistochemistry showed that apoptosis occurred more in the liver area affected by both PVE and HVE than in that affected by PVE alone. CONCLUSIONS: Preoperative sequential application of PVE and HVE seems to be safe and effective in facilitating contralateral liver regeneration by inducing more severe liver damage than PVE alone. PMID- 19300229 TI - Quantitative RT-PCR detection of tumor cells in sentinel lymph nodes isolated from colon cancer patients with an ex vivo approach. AB - OBJECTIVE: To investigate quantitative RT-PCR-based detection of tumor cells in lymph nodes (LNs) isolated from colon cancer patients by ex vivo sentinel lymph node (SLN) mapping. SUMMARY BACKGROUND DATA: Although lymph node status is among the strongest prognostic factors in colon cancer patients, 20% to 30% of node negative patients experience disease recurrence. These patients may have LN metastases that are not detected by routine examination. METHODS: Ex vivo SLN mapping was applied to 131 prospectively recruited patients undergoing curative surgery for primary colon cancer. The SLNs were analyzed for the presence of tumor cells by routine histology and real-time RT-PCR quantitation of cytokeratin 20 (CK20) and mucin 2(MUC2) mRNA. RESULTS: SLNs were identified in 125 (95%) of the 131 patients included.Routine histologic analysis of SLNs and other regional lymph nodes revealed LN metastases in 42 patients (N+), of which 29 (69%) had metastases detected in 1 or more SLNs (sensitivity, 69%; false negative rate, 31%).When analyzing the SLNs by quantitative RT-PCR, the sensitivity, compared with routine LN examination, was 37/42 (88%) for both the CK20 and the MUC2 mRNA markers. In addition, 46% and 27% of the patients' node negative by routine LN examination (N0) were positive for the CK20 and MUC2 mRNA markers, respectively, possibly reflecting the presence of occult tumor cells in their SLNs. CONCLUSIONS: Quantitative RT-PCR analysis of SLNs identified N+ patients with high sensitivity and revealed a subgroup of N0 patients with potential occult LN disease. PMID- 19300230 TI - Laparoscopy decreases postoperative complication rates after abdominal colectomy: results from the national surgical quality improvement program. AB - OBJECTIVE: Compare outcomes of non-emergent laparoscopic to open colon surgery. BACKGROUND: Laparoscopy has revolutionized much of gastrointestinal surgery. Colon and rectal surgery has seen drastic changes with many of the abdominal operations being performed laparoscopically. However, data comparing recovery and complications in patients undergoing laparoscopic and open colon surgery has shown only slight benefits for laparoscopy. Given the large benefits of laparoscopy in most gastrointestinal surgical procedures, this outcome is surprising. We, therefore, have set out to test the hypothesis that laparoscopic approaches decreases postoperative complications. METHODS: We have undertaken a review of the database maintained by the American College of Surgeon's National Surgical Quality Improvement Program. We have identified 8660 patients who met inclusion criteria for this study. Postoperative complication data were collected for patients undergoing laparoscopic or open colon surgery. Using a combination of univariate and multivariate analyses we evaluated for statistical significance. RESULTS: We found that laparoscopy decreased overall complications as well as individual complications. We found a decreased length of stay as well as a decreased risk for postoperative complications in the elderly. We found that laparoscopy decreased complication rate independent of the probability of morbidity statistic. CONCLUSIONS: When controlled for probability of morbidity, laparoscopy decreases the rate of postoperative complications. Given the equivalent outcomes of laparoscopic approaches, we conclude that laparoscopy should be offered to all patients who lack an absolute contraindication for laparoscopic surgery. PMID- 19300231 TI - Intraperitoneal treatment for peritoneal mucinous carcinomatosis of appendiceal origin after operative management: long-term follow-up of the Mayo Clinic experience. AB - OBJECTIVE: To determine prognostic factors and the impact of intraperitoneal (IP) treatment after surgical resection of peritoneal mucinous carcinomatosis (PMC) of appendiceal origin. SUMMARY OF BACKGROUND DATA: PMC is a rare, malignant, intra abdominal neoplasm that produces large amounts of mucin. Patients typically present with diffuse peritoneal disease. After surgical treatment, multiple locoregional recurrences are common; recurrences outside the abdomen are infrequent. Treatment regimens include debulking, radiotherapy with IP radioisotopes, and chemotherapies (IP, systemic, or both). Because reported data are variable and heterogeneous, treatment evaluations are challenging. METHODS: We retrospectively reviewed 115 consecutive patients with PMC who underwent maximal surgical resection with or without postoperative therapy between 1985 and 2000 at Mayo Clinic Rochester. After maximal resection, 37 patients received IP 5 fluorouracil, 35 of whom also received IP chromic phosphate P 32. The Kaplan Meier method was used to estimate overall survival (OS) and disease-free survival. RESULTS: All gross disease was removed in 61% of patients. With a median follow-up of 6.1 years, the median OS was 8.1 years. Median OS for patients receiving versus not receiving IP therapy was 23.5 years versus 7.5 years, respectively. The 5-, 10-, and 15-year OS for those receiving and not receiving IP therapy was 82%, 65%, and 52% versus 60%, 27%, and 15%, respectively. Adverse prognostic factors for OS identified by univariate analysis included partial mucin debulking, adenocarcinoma histology, systemic chemotherapy, diffuse IP disease at presentation, and no IP therapy. On multivariate analysis, diffuse IP disease at presentation and no IP therapy remained significant. A separate analysis was performed for the 70 patients who underwent gross total resection, 51% of whom received IP therapy. Adverse prognostic factors for OS included adenocarcinoma histology, systemic chemotherapy, and no IP therapy. CONCLUSIONS: This large, single-institution, retrospective series with long-term follow-up suggests that IP chromic phosphate P 32 and 5-fluorouracil after maximal surgical resection of PMC of appendiceal origin is associated with improved OS and disease-free survival. PMID- 19300232 TI - Randomized, controlled, prospective trial of the use of a mesh to prevent parastomal hernia. AB - BACKGROUND: The prevalence of terminal parastomal hernia (PH) after colostomy placement may be as high as 50%. The effect of the PH may range from discomfort to life-threatening complications. Surgical procedures for repairing PH are difficult to perform and present a high-failure rate. OBJECTIVE: To reduce the incidence of PH by implanting a lightweight mesh in the sublay position. MATERIAL AND METHODS: Randomized, controlled, prospective study. Patients were scheduled for permanent end colostomy surgery to treat cancer of the lower third of the rectum, performed by the same colorectal surgery team. An Ultrapro lightweight mesh was inserted in the sublay position in the study group. Using simple randomization, the sample size required was estimated to be 27 per group. Patients were followed-up clinically and radiologically with abdominal computed tomography by an independent clinician and a radiologist who were all blind to the aims of the study, 1 month and every 6 months after surgery. RESULTS: : The groups were homogeneous in terms of their clinical and demographic characteristics. Surgical time and postoperative morbidity were similar in the 2 groups. Mortality was 0. No mesh intolerance was reported. In the clinical follow up (median: 29 months, range: 13-49), 11/27 (40.7%) hernias were recorded in the control group compared with 4/27 (14.8%) in the study group (P = 0.03). Abdominal computed tomography identified 14/27 (44.4%) hernias in the control group compared with 6/27 (22.2%) in the study group (P = 0.08). CONCLUSIONS: Parastomal placement of a mesh reduces the appearance of PH. The technique is safe, well tolerated, and does not increase morbidity rates. PMID- 19300233 TI - Interrupted or continuous slowly absorbable sutures for closure of primary elective midline abdominal incisions: a multicenter randomized trial (INSECT: ISRCTN24023541). AB - OBJECTIVE: In patients undergoing midline incisions, the abdominal fascia can be closed with a continuous or interrupted suture using various materials. The aim of this study is to compare: (1) interrupted technique with rapidly absorbable sutures and (2) continuous techniques with different slowly absorbable sutures, focusing on the incidence of incisional hernias within 1 year. SUMMARY OF BACKGROUND DATA: A meta-analysis suggested that the incidence of incisional hernias can be more effectively reduced with slowly absorbable continuous sutures. METHODS: Multicenter randomized surgical trial with 3 parallel groups. Patients were scheduled for primary elective midline incisions. All surgeons were trained (4:1 suture wound length in continuous groups) and monitored. Primary end point, measured within 1 year after surgery, was the frequency of incisional hernias diagnosed by clinical examination and confirmed by ultrasound. Complications and safety were used as secondary end points. This study has been registered with the ISRCTN Register (INSECT: ISRCTN24023541). RESULTS: Conducted on 625 randomized patients (210 interrupted Vicryl, 205 continuous polydioxanone suture (PDS), 210 continuous Monoplus), the primary analysis showed an incidence of 28 incisional hernias (15.9%) versus 15 (8.4%) versus 22 (12.5%) for the 3 closure techniques, respectively (P = 0.09). No significant difference was observed between the 3 groups with regard to burst abdomen (4 [2.0%] vs. 6 [3.0%] vs. 8 [4.0%], P = 0.46), wound infection (26 [12.7%] vs. 39 [19.4%] vs. 33 [16.3%], P = 0.19), pulmonary infections (9 [4.4%] vs. 5 [2.5%] vs. 5 [2.5%], P = 0.46), serious adverse events (63 [30.0%] vs. 57 [27.8%] vs. 61 [29.1%], P = 0.89), and 1-year mortality (16 [7.9%] vs. 11 [5.5%] vs. 16 [7.9%], P = 0.54). CONCLUSIONS: The incidence of incisional hernias and the frequency of wound infection was higher than expected in all groups. New concepts need to be developed and studied to substantially reduce the frequency of incisional hernias. PMID- 19300234 TI - Regional collaborations as a tool for quality improvements in surgery: a systematic review of the literature. AB - BACKGROUND: A systematic review of the literature identifying regional collaborations in surgical practice examining practices related to quality improvement. METHODS: The MEDLINE, EMBASE, and Cochrane Library databases, were searched for published reports of regional collaborations in the surgical community relating to initiatives to enhance quality improvement, quality of care, patient safety, knowledge transfer, or communities of practice. RESULTS: Seven collaborative initiatives met the inclusion criteria and were included in the systematic review of the evidence. Motivations for initiating collaborations were often in response to external demands for performance data. Changes in the processes of clinical care and improvements in clinical outcomes were reported on the basis of the collaborative efforts. Significant improvements in clinical outcomes such as decreases in mortality rates, lower duration of postoperative intubations, and fewer surgical-site infections were reported. Quality improvement process measures were also reported to be improved across all of the collaborative initiatives. Success factors included (a) the establishment of trust among health professionals and health institutions; (b) the availability of accurate, complete, relevant data; (c) clinical leadership; (d) institutional commitment; and (e) the infrastructure and methodological support for quality management. CONCLUSIONS: A community of practice framework incorporating the success elements described in the systematic review of the literature can be used as a valuable model for collaboration amongst surgeons and healthcare organizations to improve quality of care and foster continuing professional development. PMID- 19300235 TI - Antibiotic prophylaxis at urinary catheter removal prevents urinary tract infections: a prospective randomized trial. AB - OBJECTIVE: To assess whether antibiotic prophylaxis at urinary catheter removal reduces the rate of urinary tract infections. SUMMARY OF BACKGROUND DATA: Urinary tract infections are among the most common nosocomial infections. Antibiotic prophylaxis at urinary catheter removal is used as a measure to prevent them, albeit without supporting evidence. METHODS: A prospective randomized study enrolled 239 patients undergoing elective abdominal surgery, who were randomized either for receiving 3 doses of trimethoprim-sulfamethoxazole at urinary catheter removal, or not. Urinary tract infections were diagnosed according to Center of Disease Control definitions. Urinary cultures were obtained before and 3 days after catheter removal. Subjective symptoms were assessed by an independent study blind urologist. RESULTS: Patients who received antibiotic prophylaxis showed significantly fewer urinary tract infections (5/103, 4.9%) than those without prophylaxis (22/102, 21.6%), P < 0.001. The absolute risk reduction for the occurrence of a urinary tract infection was 16.7%; the relative risk reduction was 77.5%, and the number needed to treat was 6. Patients with antibiotic prophylaxis also had less significant bacteriuria 3 days after catheter removal (17/103, 16.5%) than those without (42/102, 41.2%), P < 0.001. CONCLUSIONS: Antibiotic prophylaxis with trimethoprim-sulfamethoxazole on urinary catheter removal significantly reduces the rate of symptomatic urinary tract infections and bacteriuria in patients undergoing abdominal surgery with perioperative transurethral urinary catheters. PMID- 19300236 TI - Is antibiotic prophylaxis in surgery a generally effective intervention? Testing a generic hypothesis over a set of meta-analyses. AB - CONTEXT: While the main focus of a meta-analysis is often to assess the effectiveness of a particular intervention in managing or curing a specific condition, there exists a substantial amount of information within published systematic reviews that could be used to assess the validity of a generic hypothesis about the effectiveness of an intervention across a range of different but related conditions. OBJECTIVE: To systematically search for and then collate the results of meta-analyses for the effectiveness of antibiotic prophylaxis in preventing postoperative wound infection across various types of surgery in order to assess generic hypotheses about the effectiveness of this intervention in surgery as a whole. With the relative risk of wound infection used as the measure of clinical effectiveness, the hypotheses to be assessed were first that antibiotic prophylaxis would be an effective intervention for preventing wound infection over a broad range of different surgical procedures, and second that there would be a substantial difference in the effectiveness of antibiotic prophylaxis between "clean" and "contaminated" surgical procedures. DATA SOURCES: Medline and the Cochrane Database of Systematic Reviews. STUDY SELECTION: Eligible meta-analyses were meta-analyses published between 1990 and 2006 of randomized controlled trials that looked at the effectiveness of prophylactic antibiotics versus no antibiotic or placebo in preventing postoperative wound infections. DATA EXTRACTION: Independent data extraction by multiple observers. RESULTS: The first hypothesis was strongly supported by the data as evidenced by the fact that the estimates of the relative risk of infection for the 23 types of surgery that were included in the study were all less than 1. However, there was no real evidence supporting the second hypothesis that the relative risk of wound infection would substantially vary over different levels of surgery cleanliness. CONCLUSION: : As well as antibiotic prophylaxis being a generally effective intervention for preventing postoperative wound infection, the level of this effectiveness would appear to be reasonably independent of what type of surgery is being considered. Therefore, the general prevailing attitude that antibiotic prophylaxis should be assumed to be ineffective unless its effectiveness has been experimentally proven beyond doubt for the specific type of surgery being considered, perhaps should be revised. In particular, perhaps a sensible philosophy would be to assume that antibiotic prophylaxis is effective in reducing the risk of wound infection for all types of surgery, even ones where no clinical trial data exists and make exceptions to this rule if, for certain types of surgery, it can be proved to the contrary. PMID- 19300237 TI - Should total number of lymph nodes be used as a quality of care measure for stage III colon cancer? AB - OBJECTIVE: To assess whether TNODS is an independent prognostic factor after adjusting for the lymph node ratio (LNR). SUMMARY BACKGROUND DATA: The medical literature has suggested that the TNODS is associated with better survival in stage II and III colon cancer. Thus TNODS was endorsed as a quality measure for patient care by American College of Surgeons, National Quality Forum. There is, however, little biologic rationale to support this linkage. METHODS: : A total of 24,477 stage III colon cancer patients were identified from Surveillance, Epidemiology, and End Results cancer registry and categorized into 4 groups, LNR1 to LNR4, according to LNR interval: <0.07, 0.07 to 0.25, 0.25 to 0.50, and >0.50. Patients were also stratified according to TNODS into high TNODS (> or = 12) and low TNODS (<12) groups. The method of Kaplan-Meier was used to estimate the 5 year survival and the log-rank test was used to test the survival difference among the different groups. RESULTS: Patients with high TNODS have better survival compared with those with low TNODS (5-year survival 51.0% vs. 45.0%, P < 0.0001). However, after stratifying by LNR status, there was no significant survival difference between patients with high TNODS and those with low TNODS within strata LNR2 (5-year survival 56.3% vs. 56.0%, P = 0.26). Ironically, patients with high TNODS had significantly worse survival than those with low TNODS within strata LNR3 (5-year survival 41.2% vs. 47.4%, P = 0.0009) and LNR 4 (5-year survival 22.0% vs. 32.1%, P < 0.0001). CONCLUSIONS: The previously reported prognostic effect of TNODS on node-positive colon cancer was confounded by LNR. This observation calls into question the use of TNODS as a quality measure for colon cancer patients' care. PMID- 19300238 TI - Antidepressant efficacy screening of novel targets in the chick anxiety depression model. AB - The chick anxiety-depression model is a hybrid simulation, which may prove useful as an early preclinical dual pharmacological screen for novel therapeutics. Separate dose-response studies were conducted with seven test compounds that have screened positive for antidepressant effects in rodent depression models and included prasterone (5.0-40.0 mg/kg), memantine (2.5-20.0 mg/kg), ketamine (1.0 10.0 mg/kg), mifepristone (50.0-400.0 mg/kg), DOV216,303 (5.0-20.0 mg/kg), CGP36742 (2.5-15.0 mg/kg), and antalarmin (1.0-30.0 mg/kg). Chicks aged 4-6 days posthatch received test compounds intramuscularly 15 min before social separation, in which distress vocalization rates were recorded. High rates of vocalization in the first phase (0-5 min) of social separation seem to model an anxiety-like state and lower rates of vocalization in the second phase (30-60 min) seem to model a depression-like state. Prasterone, memantine, ketamine, and DOV216,303 attenuated and CPG36742 enhanced the pattern of vocalizations in the first phase. Prasterone, ketamine, mifepristone, DOV216,303, and CPG36742 attenuated and memantine and antalarmin enhanced the pattern of vocalizations in the second phase. This pattern of drug effects parallels what clinical data exist, and highlights two important characteristics of this dual-screening assay. For the compounds tested, this chick model identified phase II and III clinical failures (e.g. memantine and antalarmin) and has the potential to reveal possible contraindications of compounds (i.e. CPG36742) in cases where anxiety symptoms are concomitant with a depressive episode. PMID- 19300239 TI - Atmospheric pressure changes and unexplained variability in INR measurements. AB - Changes in atmospheric pressure may influence hepatic blood flow and drug metabolism. Anecdotal experience suggests international normalized ratio (INR) variability may be temporally related to significant atmospheric pressure changes. We investigated this potential association in a large sample of patients with multiple INRs. This is a retrospective review of outpatient anticoagulation records from the Iowa City Veteran's Affairs Medical Center and affiliated outpatient clinics from October 1999 to July 2007. All patients, receiving at least one prescription for warfarin and INR at least 30 days or more from the date of the first warfarin prescription, were identified. INRs during periods of hospitalization and vitamin K use were excluded. Proximity analysis using geocoding of ZIP codes of identified patients to the nearest National Oceanic and Atmospheric Administration station was performed to assign atmospheric pressure with INR. Spearman's Rho and Pearson's correlation were used to evaluate atmospheric pressure and INR. Unique patients (1441) with 45 187 INRs were analyzed. When limited to nontherapeutic INRs following a previously therapeutic INR (1121 unique patients/5256 INRs), a small but clinically insignificant association between delta INR and delta atmospheric pressure was observed (r = 0.025; P = 0.038), but not for actual INR and atmospheric pressure (P = 0.06). Delta atmospheric pressure demonstrated greater variation during fall/winter months compared with spring/summer (0.23 vs. 0.15 inHg; P < 0.001); however, variability in INRs for the corresponding seasons was not significant (P = 0.136). No significant difference was detected in the proportions of nontherapeutic INRs among the different seasons (P = 0.371). No correlation was observed between atmospheric pressure changes and INR variability. These findings refute the anecdotal experience seen in our anticoagulation clinic. PMID- 19300240 TI - 5 flourouracil-induced apical ballooning syndrome: a case report. AB - The apical ballooning syndrome (ABS) is a recently described stress-mediated acute cardiac syndrome characterized by transient wall-motion abnormalities involving the apex and midventricle with hyperkinesis of the basal left ventricular (LV) segments without obstructive epicardial coronary disease. Cardiotoxicity is not an uncommon adverse effect of chemotherapeutic agents. However, there are no reports of ABS secondary to chemotherapeutic agents. We describe the case of a woman who developed the syndrome after chemotherapy for metastatic cancer. A 79-year-old woman presented with typical ischemic chest pain, elevated cardiac enzymes with significant ST-segment abnormalities on her electrocardiogram. She underwent recent chemotherapy with fluorouracil for metastatic colorectal cancer. Echocardiography revealed a wall-motion abnormality involving the apical and periapical segments which appeared akinetic. Coronary angiography revealed no obstructive coronary lesions. The patient was stabilized with medical therapy. Four weeks later she remained completely asymptomatic. Echocardiogram revealed a normal ejection fraction and a resolution of the apical akinesis. Pathogenetic mechanisms of cardiac complications in cancer patients undergoing chemotherapy include coronary vasospasm, endothelial damage and consequent thrombus formation. In our patient, both supraphysiologic levels of plasma catecholamines and stress related neuropeptides caused by cancer diagnosis as well as chemotherapy may have contributed the development of ABS. PMID- 19300241 TI - Anaphylaxis in patients with congenital bleeding disorders and inhibitors. AB - Anaphylactic reactions are rare emergencies observed in patients with inherited bleeding disorders. When these adverse events occur in patients with inhibitors, they further complicate the management of an already challenging clinical situation. Anaphylactoid inhibitors have been reported in patients with inhibitors associated with hemophilia A, hemophilia B, and type 3 von Willebrand disease. In this review, we summarize the current literature data on the occurrence of anaphylactoid reactions in patients with inherited bleeding disorders and alloantibodies. In particular, we focus on the pathophysiology of the inhibitor development and its management. PMID- 19300242 TI - Severe bleeding and miscarriages in a hypofibrinogenemic woman heterozygous for the gamma Ala82Gly mutation. AB - We report a case of hypofibrinogenemia caused by heterozygosity for gamma Ala82Gly in a 69-year-old Polish woman with severe bleeding tendency and a history of six miscarriages. She suffered from frequent mucocutaneous bleedings, epistaxes requiring therapeutic interventions, prolonged bleedings after tooth extractions and surgical interventions. Her mother and sister had bleeding tendency. Fibrinogen levels ranged from 0.93 to 2.0 g/l (von Clauss method). Fibrinogen antigen levels determined by immunonephelometry were about 2 g/l. No other coagulation, platelet or liver function tests yielded abnormal results. Antiphospholipid syndrome was excluded. This report suggests that the fibrinogen gamma Ala82Gly mutation could have variable clinical presentation. PMID- 19300243 TI - Severe midline craniofacial anomalies: overlap with Pai syndrome. PMID- 19300244 TI - Anesthetic process, organization, management and economic issues: the French perspective. AB - PURPOSE OF REVIEW: Anesthesia is a three-step process: preoperative evaluation, the intervention itself and postanesthetic care. In France, this scheme has been legally regulated since 1994. Since then, significant progress has been made in terms of safety. Nevertheless, challenges in the delivery and financing of anesthesia services persist; in particular, demographic (patients and medical staff, as in other western countries), budgetary and organizational restrictions. RECENT FINDINGS: The poor efficiency (i.e. low utilization rate) of operating sites remains problematic. Results of published surveys yield the same observations: low productivity (anesthesia personnel present and paid but insufficiently occupied with mean occupied-time rate of approximately 60%), absence or poor planning and inadequate risk management. The failure to define policies to obtain better distribution of anesthesiologists throughout the country and to optimize operating-suite use are major barriers. In addition, the appropriate composition of anesthesia teams, for example nurses and physicians, for specific procedures remains undefined. Moreover, the number of anesthesia cases has grown constantly over the past 20 years, and in some situations they are not medically justified. Finally, pressure to increase productivity is becoming a potential threat to safety and deserves more discussion. SUMMARY: Paths to improvement of delivery of anesthesia in France may include: more optimal sharing of medical resources; better utilization of operating sites, perhaps by consolidating and reducing locations; applying improved organizational skills; and improved risk management. PMID- 19300245 TI - Coagulopathy in trauma patients: what are the main influence factors? AB - PURPOSE OF REVIEW: Coagulopathy and bleeding after severe injury is a common problem. Whenever caring for critically ill patients, clinicians must anticipate, recognize and manage the coagulopathy of trauma. When left untreated, cardiovascular shock and multiorgan system failure ensue. Uncompensated hemorrhage often culminates in death, highlighting the significance of recognizing the main influences in coagulopathy of trauma. RECENT FINDINGS: With recent improvements in prehospital care, trauma specialists face more challenging cases than ever before. Hemostatic transfusion strategies, with early and more aggressive use of plasma, platelets, cryoprecipitate and coagulation factor isolates, decrease blood loss in trauma patients. Combined with point-of-care testing for thromboelastography, coagulation panels, lactate and local pO2, there is an opportunity for frontline trauma clinicians to directly improve patient outcomes. SUMMARY: Although mortality previously was thought to be summarily independent of medical interventions and resuscitations, we now know the opposite to be true; it is our expectation and indeed our obligation to recognize and manage the coagulopathy of trauma better than in past years. In as much as we continue to prevent acidosis, hypothermia and the progressive coagulopathy following injury, trauma victims the world over are benefiting and surviving longer, living proof that demonstrates the utility of managing the coagulopathy of trauma. PMID- 19300246 TI - Farnesoid X receptor agonists in biliary tract disease. AB - The farnesoid X receptor (FXR) is a member of ligand-activated nuclear receptor superfamily. FXR is a bile sensor and is part of a complex network of nuclear receptors that includes also the constitutive androstane receptor and the pregnane X receptor. These receptors act coordinately to regulate essential steps of bile acids and xenobiotics uptake, metabolism and excretion in hepatocytes, cholangiocytes and kidney cells. Preclinical models indicate that FXR agonists are effective in reducing liver injury in nonobstructive models of cholestasis. FXR ligands are currently under investigation for treating patients with early stage primary biliary cirrhosis. Although these ligands hold promise, evidence is growing that FXR activation could impair the expression/activity of basolateral transporters such as multidrug resistance protein 4 essential for basolateral secretion of bile constituents in the systemic circulation. Because FXR, pregnane X receptor and constitutive androstane receptor ligands interact with different target genes, it appear that a combination with pregnane X receptor, constitutive androstane receptor ligand/activator or both or ursodeoxycholic acid could prevent possible side-effects of FXR activation in severe cholestasis. PMID- 19300247 TI - Early nephropathy in type 1 diabetes: the importance of early renal function decline. AB - PURPOSE OF REVIEW: The results of recent clinical trials in early diabetic nephropathy demonstrate that current therapies designed to suppress microalbuminuria do not prevent renal function decline. However, recent observational studies refined the traditional model of early nephropathy in type 1 diabetes and may inform more effective therapies for the prevention of chronic kidney disease. RECENT FINDINGS: A contemporary model of early nephropathy in type 1 diabetes has emerged in which initiation of renal function decline occurs soon after the onset of microalbuminuria and is not conditional on progression to proteinuria. Early renal function decline can be diagnosed using serial measurement of serum cystatin C. Abnormal levels of markers of protein glycation, uric acid metabolism, and chronic inflammation appear to represent mechanisms unique to early renal function decline and distinct from those involved in microalbuminuria. SUMMARY: Recent findings refine the existing paradigm of early nephropathy in type 1 diabetes and have significant implications for research. Clinical tests--such as an algorithm for the serial determination of serum cystatin C--should be developed for monitoring early renal function decline for use as an outcome in clinical trials. PMID- 19300248 TI - Pathological predictors of prognosis in immunoglobulin A nephropathy: a review. AB - PURPOSE OF REVIEW: To provide a review and discussion of histological prognostic indicators in immunoglobulin A (IgA) nephropathy (IgAN). RECENT FINDINGS: A variety of histological parameters and classifications have been used to attempt to predict prognosis in patients with IgAN. Grading systems used thus far do not consistently provide a useful adjunct to clinical prognostic parameters. This may be due to the variety of grading systems used, to inconsistent patient recruitment processes, and to the use of actuarial renal survival as an end point. This has led to the development of a new IgAN classification proposal by the International IgA Nephropathy Network in conjunction with the Renal Pathological Society. Additional potential markers of disease progression currently under investigation include glomerular parameters such as number and size, markers of podocyte function and of complement activation, inflammatory infiltrates and mediators of tubulointerstitial fibrosis. SUMMARY: There is a need for an internationally accepted, reproducible and clinically meaningful pathological classification of IgAN. Such a classification is currently being developed. By using only reproducible, single parameters and validating them on a large group of cases from all over the world, it is hoped that a useful clinically predictive tool will be developed. PMID- 19300249 TI - HIV-1 associated dementia: update on pathological mechanisms and therapeutic approaches. AB - PURPOSE OF REVIEW: Infection with HIV-1 can induce dementia despite successful administration of life-prolonging highly active antiretroviral therapy. This review will discuss recent progress toward a better understanding of the pathogenesis and an improved design of therapies for HIV-associated neurocognitive disorders. RECENT FINDINGS: Highly active antiretroviral therapy prolongs the lives of HIV patients, but the incidence of HIV-associated dementia as an AIDS-defining illness has increased and the brain is now recognized as a viral sanctuary that requires additional therapeutic effort. The neuropathology of HIV infection also has changed due to improved therapy, and while more similarities with other neurodegenerative diseases are being reported, predictive biomarkers remain elusive. However, improvements of in-vivo imaging technology and progress in uncovering the molecular mechanisms of HIV disease keep providing new insights. As such it appears that a prolonged activation of the immune system by HIV eventually leads to AIDS, and several lines of evidence indicate that simultaneously neurotoxic processes and impairment of neurogenesis both contribute to the development of HIV-associated neurocognitive disorders. SUMMARY: The improved understanding of the interaction between HIV and its human host provides hope that adjunctive therapies to antiretroviral treatment can be developed for HIV-associated neurocognitive disorders. PMID- 19300250 TI - Chronic tension-type headache: what is new? AB - PURPOSE OF REVIEW: This review discusses current data on nosological boundaries related to diagnosis, pathophysiology and therapeutic strategies in chronic tension-type headache (CTTH). RECENT FINDINGS: Diagnostic criteria of CTTH should be adapted to improve its sensitivity against migraine. It seems that mechanical pain sensitivity is a consequence and not a causative factor of CTTH. Recent evidence is modifying previous knowledge about relationships between muscle tissues and CTTH, suggesting a potential role of muscle trigger points in the genesis of pain. An updated pain model suggests that headache perception can be explained by referred pain from trigger points in the craniocervical muscles, mediated through the spinal cord and the trigeminal nucleus caudalis rather than only tenderness of the muscles themselves. Different therapeutic strategies, pharmacological, physical therapy, psychological and acupuncture, are generally used. The therapeutic efficacy of nonsteroidal anti-inflammatory drugs remains incomplete. The tricyclic antidepressants are the most used first-line therapeutic agents for CTTH. Surprisingly, few controlled studies have been performed and not all of them have found an efficacy superior to placebo. Further, there is insufficient evidence to support/refute the efficacy of physical therapy in CTTH. SUMMARY: Although there is an increasing scientific interest in CTTH, future studies incorporating subgroups of patients who will likely to benefit from a specific treatment (clinical prediction rules) should be conducted. PMID- 19300251 TI - Prematurity prevention: the role of progesterone. AB - PURPOSE OF REVIEW: This review summarizes the evidence of the effectiveness of progesterone on the rate of preterm birth and evaluates the most recent studies. RECENT FINDINGS: The incidence of preterm delivery is about 7-11% of all pregnant women and preterm birth is one of the most important causes of neonatal morbidity and mortality. Interventions to reduce such complications have been attempted for several years. Most efforts so far have been tertiary interventions, such as treatment with antenatal corticosteroids, tocolytic agents, and antibiotics. Some of these measures have reduced perinatal morbidity and mortality, but the incidence of preterm birth is increasing. Recently, researches have suggested prophylactic progesterone could reduce the preterm birth rate in a select group presenting previous preterm birth and a short cervical length by transvaginal scan at mid-trimester pregnancy. SUMMARY: This review intends to define the current indication for administration of progesterone for pregnant women. On the basis of current knowledge, progesterone should be offered to women with a documented history of a previous spontaneous birth at less than 37 weeks and for those found to have a short cervical length of 15 mm or less. Studies are needed to evaluate progesterone efficacy on other risk factors. PMID- 19300252 TI - The science and art of quality in obstetric ultrasound. AB - PURPOSE OF REVIEW: Due to the imbalance between increasing demand, limitations in healthcare resources and an increase in lawsuit cases in prenatal ultrasound practice, the need for quality has emerged in the field of prenatal ultrasound. RECENT FINDINGS: Over the past few years, various approaches for quality control based on nuchal translucency measurements, other biometric measurements or fetal images have been developed and tested. SUMMARY: This review describes various qualitative and quantitative quality control procedures. Special emphasis is put on the use of fetal biometry to develop quality control systems based on both qualitative and quantitative approaches. PMID- 19300253 TI - Effects of low-dose aspirin in in-vitro fertilization. AB - PURPOSE OF REVIEW: In theory, use of aspirin in IVF is based on its anti inflammatory, vasodilatory, and platelet aggregation inhibition properties, which improve blood flow to a woman's implantation site. It is hypothesized that this effect on blood flow will improve success rates. RECENT FINDINGS: Clinical studies investigating the use of low-dose aspirin (LDA) as an adjuvant therapy to IVF have produced conflicting results. The conflicting results have come as a consequence of the heterogeneous mixture of clinical trials with lack of adequate power. Even after multiple meta-analyses, differing estimates of effect were calculated as to whether aspirin should be used in conjunction with IVF. SUMMARY: Conflicting results leave the question of the effects of LDA in IVF unanswered. More trials are required for analysis to have adequate statistical power and until then the data remain unclear. At this point, there are not enough data to show that aspirin has a beneficial effect on the outcomes of IVF, but absence of effect is not adequate grounds to overturn the current clinical practice for those using LDA in efforts aimed at achieving success with IVF. PMID- 19300254 TI - Hemostasis and coagulation monitoring and management during liver transplantation. AB - PURPOSE OF REVIEW: The objective of this review is to evaluate the present state of nonpharmacological measures, pharmacological measures applied to reduce bleeding and perioperative blood products management during orthotopic liver transplantation. RECENT FINDINGS: Recent studies improve the knowledge in the hemostatic response in cirrhotic patients such as thrombin generation and platelet adhesion due to elevated levels of von Willebrand factor. Restrictive fluid therapy is an important measure to avoid hemodilution and so bleeding. Prophylaxis with antifibrinolytics is being questioned for the risk of thrombosis. Correction of coagulation defects with fresh frozen plasma has not reduced blood loss, and it has been related to worse outcome. Also, platelets administration has a negative effect in the outcome. SUMMARY: In order to maintain hemostatic system in compensated cirrhotic patient, every effort we do to improve it must consider not to imbalance it resulting in thrombi-hemorrhagic events. Pharmacological measures must be based on their clinical evidence. Identification of high risk bleeding patients would help in developing coagulation guidelines. PMID- 19300255 TI - Editorial xenotransplantation. PMID- 19300256 TI - Incentivizing living organ donation. AB - PURPOSE OF REVIEW: The number of organs available for the patients on the transplant waitlist remains at a disproportionate low. All possible methods to curtail this shortage, including providing donors with incentives, have been proposed. This article reviews recent publications addressing the benefits and risks involved in incentivizing living donation. RECENT FINDINGS: The debate about the ethics, feasibility, and possible models for compensating organ donors has been prominent in recent literature. As certain countries take lead on this initiative, others are cautiously weighing in on the impact implementations of such policies may have on the society, especially on the underprivileged. SUMMARY: The shortage of organs has resulted in proposal of strategies that encroach on certain moral and ethical principles. Providing incentives to donors is one such strategy that is likely to receive a lot of attention in the next few years. PMID- 19300257 TI - Cash, rewards, and benefits in organ transplantation: an open letter to Senator Arlen Specter. AB - PURPOSE OF REVIEW: To consider proposals to use financial incentives for organ donors that have become a subject of intense controversy in both lay and medical press (in contradistinction to the removal of financial disincentives, which is essentially noncontroversial although typically not practiced). RECENT FINDINGS: In a concerned response to the shortage of organs the office of Senator Specter of Pennsylvania has been the source of a proposal to amend the 1984 United States National Organ Transplant Act, which has been interpreted to prohibit such incentives. The proposal would permit various forms of financial incentives for donation to no longer be prohibited. SUMMARY: The amendment would have unintentional negative consequences that could undermine, rather than strengthen, the national and international organ transplant endeavor. These concerns are considered in my personal correspondence to Senator Specter's office on which the text is based. PMID- 19300258 TI - The implications of Istanbul Declaration on organ trafficking and transplant tourism. AB - PURPOSE OF REVIEW: Organ trafficking, transplant tourism and transplant commercialism are now defined by the Declaration of Istanbul; the Declaration provides principles of practice based on those definitions. Organ trafficking and transplant tourism should be prohibited because they violate the principles of equity, justice and respect for human dignity. RECENT FINDINGS: This report provides a country-by-country description of current events that may effect the practice of transplantation internationally for the foreseeable future. SUMMARY: The implications of the Istanbul Declaration are profound. It calls for a legal and professional framework in each country to govern organ donation and transplantation activities. It calls for a transparent regulatory oversight system that ensures donor and recipient safety and enforces the prohibitions of unethical practices. Governments should ensure the provision of care and follow up of living donors be no less than the care and attention provided for transplants recipients. PMID- 19300259 TI - Immune responses to alpha1,3 galactosyltransferase knockout pigs. AB - PURPOSE OF REVIEW: To summarize the current knowledge of the immune response generated against xenografts stemming from alpha1,3-galactosyltransferase knockout (GalT-KO) pigs. In particular, we will address the nature of potentially remaining Gal epitopes, the role of non-Gal xenoantigens, and the cellular response directed against GalT-KO tissues. RECENT FINDINGS: New findings support the view that porcine cells do not express isoglobotrihexosylceramide 3, and GalT KO pigs, if at all, express negligible levels of Gal. The anti-non-Gal antibody response to GalT-KO cells allowed the identification of several potentially relevant porcine xenoantigens, mainly carbohydrates. Coculture of wildtype pig aortic endothelial cells but not of GalT-KO pig aortic endothelial cells with whole human blood induces the secretion of porcine and human cytokines and the upregulation of E-selectin; in contrast, the transmigration of human leukocytes across porcine endothelium is not regulated by Gal. SUMMARY: New immunological problems are arising after the elimination of Gal by the generation of GalT-KO pigs; these include non-Gal antibodies and the identification of their elusive antigens, as well as cellular components of the immune system, including neutrophils, macrophages, natural killer cells, and T cells. PMID- 19300260 TI - Room air or 100% oxygen for resuscitation of infants with perinatal depression. AB - PURPOSE OF REVIEW: The International Liaison Committee on Resuscitation (ILCOR) recommends initiating neonatal resuscitation with concentrations of oxygen between 21 and 100%. This wide range of oxygen concentrations recommended for resuscitation highlights the lack of evidence supporting either 21 or 100% O2. The purpose of this review is to analyze the efficacy of reoxygenation with 100% O2 or room air on rates of return of spontaneous circulation--the main goal of cardiopulmonary resuscitation. RECENT FINDINGS: Clinical studies suggest that reoxygenation initiated with room air is effective in depressed neonates born with a preserved circulation. Reoxygenation with room air in these infants is associated with lower levels of circulating markers of oxidative stress than reoxygenation with 100% oxygen. However, there is no evidence that resuscitation with room air is as effective as that with 100% oxygen in restoration of an arrested circulation. In fact, animal studies indicate that, in comparison with 100% oxygen, reoxygenation with room air results in more sluggish restoration of depressed cerebral and systemic circulations. SUMMARY: Prior to a revision of current neonatal resuscitation guidelines it must be determined whether resuscitation initiated with room air results in the same rate of return of spontaneous circulation as resuscitation initiated with 100% oxygen. PMID- 19300261 TI - Antivascular endothelial growth factor for retinopathy of prematurity. AB - PURPOSE OF REVIEW: This review will discuss a potentially more effective treatment for retinopathy of prematurity (ROP) with fewer acute and long-term complications. Avastin (bevacizumab) therapy is a promising anti-vascular endothelial growth factor (anti-VEGF) administered directly into the vitreous. RECENT FINDINGS: Recent reports detail the use of Avastin alone, and in combination with light amplification by stimulated emission of radiation (LASER) therapy and vitrectomy, for ROP stages 3, 4, and 5. Currently, one clinical trial is studying Avastin alone for acute vision-threatening ROP stage 3 in zone I and posterior zone II without LASER therapy. Another clinical trial is investigating Avastin following LASER therapy for recurrent ROP stages 4 and 5. SUMMARY: Treatment for ROP has evolved from later, more destructive (cryotherapy) to earlier, less destructive (LASER therapy) peripheral retinal ablation. If evidence-based data supports early findings, the use of Avastin may be recommended without the need for ablative LASER therapy and before retinal detachment develops. Avastin will be especially useful for ROP stage 3 cases with hemorrhage decreasing retinal visualization, rigid pupils, intravitreal neovascularization with early or developing (minimal) fibrous membranes, or aggressive posterior retinopathy of prematurity (AP-ROP). These cases all continue to have poor outcomes with LASER therapy. PMID- 19300262 TI - Moving towards making social toxins mainstream in children's environmental health. AB - PURPOSE OF REVIEW: Although traditional disciplinary research theory and methods have focused separately on how social and physical environmental factors affect children's health, evolving research underscores important integrated effects. RECENT FINDINGS: This review outlines the specific reasons why social determinants should be considered mainstream in children's environmental health research with particular focus on interactive effects between social and physical hazards. These include sensitivity of overlapping physiological systems, via epigenesis, programming, and plasticity to social and physical environmental moderation that may impact health across the life span; ways in which social environmental vulnerabilities moderate the effects of physical environmental factors, providing specific examples related to respiratory health and neurodevelopment; overlapping exposure distribution profiles; and relevance to pediatric health disparities. SUMMARY: Because of the covariance across exposures, and evidence that social stress and other environmental toxins (e.g., pollutants, tobacco smoke) may influence common physiological pathways (e.g., oxidative stress, proinflammatory immune pathways, autonomic disruption), understanding the potential synergistic effects promises to more completely inform children's environmental health risk. Although this discussion focuses around the respiratory and neurological systems, these concepts extend more broadly to children's psychological and physical development. PMID- 19300263 TI - Update on child maltreatment. AB - PURPOSE OF REVIEW: The authors explore the literature published in the past year addressing child maltreatment issues, including sexual abuse, physical child abuse, inflicted head trauma, and child abuse prevention. RECENT FINDINGS: The body of knowledge about child abuse and its mimics continues to expand. Evident in this year's literature is the challenge which the diagnosis of child abuse creates for clinicians. Although further strides are being made toward universal education of providers, it is clear that there is still a reluctance to report abuse to child welfare agencies. The legal repercussions of diagnosing abuse can be extensive, and there has been a proliferation of medical defense experts who disagree with the commonly accepted tenets of abusive injury and who are vocal in the literature. SUMMARY: It remains the responsibility of pediatric providers to consider child maltreatment in the differential diagnosis of any unexplained injury or medical problem. Several studies document the high rate of spanking, slapping or shaking children, and primary care clinicians may be the first professionals in a position to begin the evaluation for possible child maltreatment. Despite the natural hesitancy to diagnose abuse, clinicians have an ethical and moral obligation to address this issue both in their practice and in their communities. The short-term and long-term costs to individuals who experience family violence have been well demonstrated and include not only emotional repercussions, but also chronic health conditions, which result in significant cost to society. PMID- 19300264 TI - Primary ciliary dyskinesia: improving the diagnostic approach. AB - PURPOSE OF REVIEW: The diagnosis of primary ciliary dyskinesia (PCD) has relied on analysis of ciliary motility and ultrastructure; however, these tests are not readily available and have not been standardized. Consequently, the diagnosis of PCD may be delayed or missed or made incorrectly. This review outlines the potential utility of new diagnostic tests, including measurement of nasal nitric oxide production and systematic analysis for mutations in genes encoding ciliary proteins. RECENT FINDINGS: Clinical manifestations of PCD have been expanded to include neonatal respiratory distress and heterotaxy. Measurement of nasal nitric oxide has emerged as a useful screening test for PCD based on the very low levels in PCD (approximately 1/10 of normal values). Genetic testing is emerging for PCD and demonstrates extensive genetic heterogeneity. Some genes and gene mutations involved in PCD have been defined. Approximately one-third of PCD cases have identifiable gene mutations in one of six different genes. An international effort is focused on defining PCD-causing defects in other genes. SUMMARY: The incorporation of nasal nitric oxide measurement as a screening test to define probable PCD cases and gene mutation analysis to make a definitive diagnosis of PCD should enhance diagnostic evaluation of PCD. PMID- 19300265 TI - Dietary intervention strategies to modulate prostate cancer risk and prognosis. AB - PURPOSE OF REVIEW: There is increasing interest in complementary and holistic approaches for cancer prevention and management. We sought to review the latest literature regarding dietary interventions for prostate cancer with a special emphasis on dietary fat and carbohydrate intake for modulating prognosis among men with prostate cancer. RECENT FINDINGS: Several recent prospective trials have investigated various dietary and lifestyle investigations on malignant prostate tissue biology. These interventions included a very low-fat (12% fat kcals) vegan diet with various supplements and lifestyle changes, a more traditional low-fat diet (25% fat kcals) with flaxseed supplementation, and a low-glycemic index diet. Low-glycemic index and very low-fat vegan diets (with supplements and lifestyle changes) alter tumor biology as assessed by tumor gene expression changes, with a common mechanism perhaps being weight loss whereas no effects were seen with a traditional low-fat diet. In mice, either very low-fat or low carbohydrate diets significantly slow tumor growth independent of weight loss. Epidemiologic and preclinical data also suggest cholesterol intake and serum cholesterol levels may be linked with the development and progression of prostate cancer. SUMMARY: Small clinical trials suggest that tumor biology can be altered by either a vegan low-fat diet or eliminating simple carbohydrates accompanied by weight loss. Larger and longer term studies are needed to determine the clinical relevance of these findings. PMID- 19300266 TI - Quality of life outcomes following treatment for localized prostate cancer: is there a clear winner? AB - PURPOSE OF REVIEW: The majority of men treated for localized prostate cancer are cured of their disease. As a result, it is important to discuss long-term quality of life (QoL) expectations when counseling patients regarding treatment options. The varying QoL outcomes for radical prostatectomy, external beam radiotherapy, brachytherapy, and cryotherapy will be reviewed. RECENT FINDINGS: Robotic and radical prostatectomy has similar outcomes with significant initial worsening of urinary continence and sexual function. External beam radiation has less impact on continence and sexual function but noteworthy bowel toxicity. Brachytherapy results in the most irritative urinary symptoms, with decreased sexual and bowel QoL as well. Cryotherapy greatly reduces sexual function. SUMMARY: Every patient has unique pretreatment variables, priorities, and preferences. It is crucial to fully explain the range of oncologic and QoL implications when counseling patients regarding treatment for localized prostate cancer. PMID- 19300267 TI - Comparison of etomidate-remifentanil and propofol-remifentanil sedation in patients scheduled for colonoscopy. AB - OBJECTIVE: The aim of the study was to compare haemodynamic responses, recovery and discharge times, and physician satisfaction of etomidate-remifentanil and propofol-remifentanil combinations in patients undergoing elective colonoscopy. METHODS: Sixty patients, aged 18-65 years, scheduled for elective colonoscopy under sedation were prospectively randomized, double blind for the study. Two minutes after the beginning of a continuous remifentanil infusion (0.1 microg kg( 1) min(-1)), etomidate or propofol were administered. Patients in the etomidate group received a 0.05 mg kg(-1) maintenance dose of etomidate after an initial dose of 0.1 mg kg(-1), and patients in the propofol group received a 0.25 mg kg( 1) maintenance dose of propofol after an initial dose of 0.5 mg kg(-1) in order to have a Ramsay sedation score of 3-4. Basal values of heart rate, mean arterial pressure, oxygen saturation, respiratory rate and Ramsay sedation score were recorded. Values were recorded every 2 min for the first 10 min and every 5 min thereafter, until the completion of the procedure. RESULTS: Mean arterial pressure was lower at 4, 6, 8, 10, 15, 20 and 25 min in the propofol group (P = 0.001). Mean respiratory rate in the propofol group at 6, 8, 10, 15, 20, 25 min was also lower (P < 0.05). The incidence of apnoea and hypotension was significantly lower in the etomidate group (P < 0.001). Arrival time into the postoperative care unit and recovery time were shorter in the etomidate group (P = 0.001, P = 0.01, respectively). Physician satisfaction in both groups was similar. CONCLUSION: Etomidate-remifentanil administration for sedation and analgesia during colonoscopy resulted in more stable haemodynamic responses and shorter recovery and discharge times. PMID- 19300269 TI - Paediatric perioperative cardiac arrest and its mortality: database of a 60-month period from a tertiary care paediatric centre. AB - BACKGROUND AND OBJECTIVE: The analysis of perioperative cardiac arrest and mortality remains a potentially valuable method to improve clinical outcome. This survey evaluated the incidence and causes of perioperative cardiac arrests and mortality in a paediatric surgical population over 5 years. METHODS: All cardiac arrests that occurred between April 2003 and March 2008, during administration of anaesthesia at the paediatric centre of our institute, were recorded from an anaesthesia database and postsurgical ICU census register. All surgery performed under anaesthesia was included whether emergency or elective. All surgical procedures were covered except eye surgery, cardiac surgery and the procedures performed at remote locations. Data collected included patient characteristics, surgical procedures, preoperative physical status and anaesthesia-provider information, immediate cause of cardiac arrest, antecedent events, management and outcome. All cardiac arrests were grouped according to the cause of arrest into one of four groups: totally anaesthesia related, partially anaesthesia related, surgery related or child condition related. RESULTS: There were a total of 27 cardiac arrests out of 12 158 procedures. Major risk factors for cardiac arrests were children under 1 year of age (P < 0.05), ASA physical status (ASA-PS) III or more (P < 0.001) and emergency surgery (P < 0.01). There were nine cardiac arrests attributed to anaesthesia, three totally and six partially related to anaesthesia. The main causes of anaesthesia-related cardiac arrest were respiratory events (56%), followed by cardiac events (33%). Anaesthesia-related mortality was 1.2/10 000 anaesthetics in patients with ASA-PS I-II and 7.7/10 000 anaesthetics in patients with ASA-PS III-V with a survival rate of 56%. CONCLUSION: Major risk factors for cardiac arrests were age under 1 year, poor physical status and emergency surgery. Respiratory and cardiovascular-related events accounted for most of all anaesthesia-related cardiac arrests. High ASA-PS was the leading cause of perioperative mortality. PMID- 19300270 TI - Cerebral state index vs. bispectral index during sevoflurane-nitrous oxide anaesthesia. AB - BACKGROUND AND OBJECTIVE: The cerebral state index (CSI) is a low-cost and compact electroencephalographic monitor alternative to the bispectral index (BIS). Although both indices are shown as the number between 0 and 100, owing to the different calculation algorithms, both indices might be different during anaesthesia. The present study was performed to compare the BIS and CSI during sevoflurane-nitrous oxide anaesthesia. PATIENTS AND METHODS: Forty patients scheduled for abdominal surgery aged 30-70 years were enrolled. The electrodes of both the cerebral state monitor and BIS were attached simultaneously. Anaesthesia was induced with midazolam and fentanyl, and was maintained with sevoflurane with 50% nitrous oxide in oxygen and intermittent epidural mepivacaine. During surgery, end-tidal sevoflurane concentration was kept at 0.5, 1.0, and 1.5% for more than 10 min to measure CSI and BIS. RESULTS: The duration of signal disturbance by electric cautery was 12.6 +/- 4.2 min in the BIS and 3.9 +/- 1.8 min in the cerebral state monitor (P < 0.05) for 9.5 +/- 4.3 min duration of electric cautery. The recovery time from disturbance by electric cautery was 37 +/- 12 s in the BIS and 4 +/- 2 s in the cerebral state monitor (P < 0.05). The absolute values of the BIS and CSI were not significantly different. Both BIS and CSI decreased significantly when end-tidal sevoflurane concentration increased from 0.5 to 1.0%, but did not significantly change when end-tidal sevoflurane concentration increased from 1.0 to 1.5%. The BIS and CSI correlated well. The Bland-Altman plot showed good agreement between the BIS and CSI. CONCLUSION: During sevoflurane-nitrous oxide anaesthesia, the CSI was not significantly different from the BIS in the awake state and with sevoflurane concentration 0.5 1.5%. PMID- 19300271 TI - A useful device for difficult nasal tracheal intubation in China. PMID- 19300272 TI - The Bonfils intubation fibrescope: clinical evaluation and consideration of the learning curve. PMID- 19300273 TI - Antireflux 'barriers': problems with patient recruitment for a new endoscopic antireflux procedure. AB - BACKGROUND: Most first-generation endoscopic antireflux procedures (EARPs) have been withdrawn because of variable success rates, economic considerations, and/or complications. As a result, subsequent methods may meet 'skepticism' by physicians and patients. AIMS: To identify potential barriers to patient recruitment for a new EARP METHODS: We prospectively analyzed our recruitment for a phase 2 study of a transoral incisionless fundoplication procedure. We contacted 50 private practices and 23 hospitals for potential referrals, and placed three newspaper advertisements. All patient replies were followed up by a phone call. Patients were then invited for a personal interview, and eligible patients underwent further preprocedure testing. In addition, poststudy questionnaires regarding their opinions about EARPs were sent to referring physicians. RESULTS: Of 134 interviewed patients, only 10% (n=13) were successfully recruited. Candidates mostly responded to newspaper advertisements (87%) or were referred from our own institution (7%). Primary exclusion criteria included failure of proton pump inhibitor response (34%), lack of proton pump inhibitor use (20%), atypical symptoms (18%), or a large hiatal hernia (17%). Seventy percent of the responding physicians did not believe that new EARPs would be superior to previous methods. CONCLUSION: The EARP market seems to be much smaller than anticipated, partially because of skepticism of referring physicians, and partially because of strict selection criteria. PMID- 19300274 TI - Minor adverse events of colonoscopy on ambulatory patients: the impact of moderate sedation. AB - BACKGROUND: Minor complications in colonoscopy, such as pain, distension, headache, dizziness, etc. are an important cause for the rejection of colonoscopy procedure. Their incidence, however, has not been established. OBJECTIVE: To determine minor complication rates in outpatient colonoscopy procedures and the effect of sedation on these complications. PATIENTS: A prospective randomized cohort study was performed on 1250 adults selected randomly among patients referred to our endoscopy unit. Complications and undesirable affects related to the colonoscopy were reordered during the patients' stay in the endoscopy unit (early complications) and 30 days postprocedure (late complications). RESULTS: One thousand one hundred and twenty-six patients were enrolled (mean age 50.43+/ 13.7 years; 54.5% female), of which 875 (78%) were sedated and 251 (22%) were not. No difference between groups was observed. Thirty-one percent of the patients had early minor complications (25% among sedated patients; 52% of nonsedated patients; P<0.001 chi2); 23% had late minor complications (16% of sedated patients vs. 51% nonsedated patients; P<0.001 chi2). The most common undesirable effects were pain and abdominal distension. The risk of experiencing complications - odds ratio - was 1.013 times higher per year of age [confidence interval (CI) 95%: 1.004-1.022]; 1.953 times higher per increase in American Society of Anesthesiologists classification (95% CI: 1.524-2.504); and 0.116 times lower when sedation was used (95% CI: 0.079-0.170). CONCLUSION: Minor complications of colonoscopy are common. Their incidence increases with age and American Society of Anesthesiologists class and decreases with the use of sedation. PMID- 19300275 TI - Faecal M2-pyruvate kinase: a novel, noninvasive marker of ileal pouch inflammation. AB - BACKGROUND: Dimeric M2-pyruvate kinase (dM2-PK) is overexpressed in tumour cells with rapid cell turnover. Its concentrations correlate well with the staging and metastatic capability of the tumour cells. We investigated the use of faecal dM2 PK as a noninvasive marker of pouch inflammation (pouchitis) in patients having undergone restorative proctocolectomy. METHODS: Stool samples were obtained from 46 patients with ulcerative colitis (UC) and eight with familial adenomatous polyposis. Pouchitis was defined using the objective pouchitis score (OPS) and the pouch disease activity index. Faecal dM2-PK was measured using a quantitative sandwich-type enzyme immunoassay (ScheBo Biotech UK) and the results compared with reciprocal faecal calprotectin concentrations. RESULTS: Using the OPS, 6 of the 46 patients with UC had pouchitis and prepouch ileitis, 13 had UC pouchitis alone, and 27 had a non-inflamed UC pouch. One patient with familial adenomatous polyposis had pouchitis and prepouch ileitis and 7 had an non inflamed pouch. Respective median dM2-PK values (U/ml) for these five groups were 49.5 (4.5-110), 12 (1-192.3), 2.2 (0.1-95.2), 19.5 and 1 (0.1-3). Statistically significant differences were noted between inflamed and non inflamed pouches (P<0.0001). dM2 PK correlated significantly with the OPS, pouch disease activity index, endoscopic appearances, acute histological and neutrophil scores (<0.0001). The receiver operating characteristic analysis demonstrated a sensitivity and specificity of 80 and 70.6%, respectively. dM2-PK and faecal calprotectin concentrations correlated closely (r=0.87, P<0.0001). CONCLUSION: This study demonstrates that faecal dM2-PK is a sensitive marker of pouch inflammation and that its concentration directly correlates with the objective markers of pouchitis severity. PMID- 19300276 TI - Flecainide as first-line treatment for supraventricular tachycardia in newborns. AB - BACKGROUND: Flecainide for the treatment of supraventricular tachycardia (SVT) in newborns is still controversial because of its potentially severe proarrhythmic effects. METHODS AND RESULTS: Between January 2004 and December 2006, we used flecainide to treat 20 consecutive newborns (15 males) with paroxysmal SVT without any structural heart disease. Their age at hospitalization was 11.5 +/- 11.1 days. The intravenous administration of flecainide (1 mg/kg) effectively restored sinus rhythm in all the patients. Once stable sinus rhythm had been restored, the drug was administered orally at a dose of 2 mg/kg/day twice daily, which was uptitrated as the patients gained weight. The patients were followed up for up to 24 months with clinical evaluations, baseline ECG and 24-h Holter monitoring every 3 months. There were neither deaths nor any episodes of heart failure or sustained ventricular tachycardia during follow-up. SVT were completely controlled in 17 patients (85%), with an oral dose of 3.35 +/- 1.35 mg/kg/day of flecainide; in the remaining three patients with refractory arrhythmias, propranolol was added for optimal treatment. No significant increase in the duration of QRS (70 +/- 1.09 vs. 63.8 +/- 1.87 ms, P = NS) or any significant QTc prolongation (413 +/- 7.4 vs. 412.6 +/- 8.01 ms, P = NS) was observed. One patient developed an incomplete right bundle branch block promptly reverted by reducing the dose. CONCLUSION: This preliminary experience indicates that flecainide is well tolerated and effective as first-line treatment for paroxysmal SVT in newborns without structural heart disease. PMID- 19300277 TI - Giant hydatid cyst of the interventricular septum mimicking acute myocardial infarction on ECG: an unusual cause of ST segment elevation. AB - Hydatid disease is a parasitic infection caused by larvae of Echinococcus granulosus, which is still endemic in many cattle-raising areas. Cardiac involvement is a rare, but potentially a very serious complication of the hydatid disease. The diagnosis of cardiac cyst hydatid may be difficult due to the nonspecific symptoms and varying clinical presentations. With this report, we describe a case of giant hydatic cyst of the interventricular septum that caused ischemic changes on ECG, mimicking acute myocardial infarction. The final diagnosis was made by combining echocardiography, MRI, and serological tests. Surgical resection of the cyst, followed by albendezol treatment yielded a favorable outcome. PMID- 19300278 TI - Coronary artery-pulmonary vein fistula diagnosed by multidetector computed tomography. AB - Coronary artery fistulas are rare congenital or acquired coronary artery anomalies that can originate from any of the three major coronary arteries and drain in all the cardiac chambers and great vessels. Multidetector computed tomography has provided cross-sectional anatomy of the heart with excellent spatial and temporal resolution, which shows the anatomical and functional changes in cardiac disorders. Here, the first case of noniatrogenic coronary artery to pulmonary vein fistula depicted by multidetector computed tomography has been presented. PMID- 19300279 TI - Effect of low-dose prednisone on leukocyte counts and subpopulations in patients with rheumatoid arthritis. PMID- 19300280 TI - Polyarticular osteochondromatosis: a rare association in rheumatoid arthritis. PMID- 19300281 TI - Rheumatologist perceptions of sources of health care disparities in minority rheumatoid arthritis patients. PMID- 19300282 TI - Devastating ochronotic arthropathy with successful bilateral hip and knee arthroplasties. AB - Ochronotic arthropathy is a manifestation of long-standing alkaptonuria, a rare hereditary metabolic disorder characterized by the absence of the enzyme homogentisic acid oxidase. We describe a patient who developed rapidly progressive destructive ochronotic arthropathy in his knee and hip joints and underwent successful bilateral total knee and hip arthroplasties. PMID- 19300283 TI - A scleroderma-like cutaneous syndrome associated with a marked Th2-type immune response occurring after a prosthetic joint implant. AB - A scleroderma-like cutaneous syndrome, occurring after implantation of a prosthetic knee joint in an elderly woman, is reported. This case did not seem to typically fit into any of the known scleroderma-like disorders of the skin described to date. The patient was shown to be sensitized to metals contained in the prosthesis and to mount a Th2-type immune response concomitantly with development of skin fibrosis. In particular, eosinophilia, markedly elevated serum IgE levels, in vitro spontaneous production of interleukin (IL)-4 by T lymphocytes, and elevated serum levels of Th2 cytokines (namely, IL-4, IL-5, and IL-13) were observed during the acute phase of illness. Since eosinophils and such Th2 cytokines as IL-13 also have recognized fibrogenic properties, it is speculated that the pathogenesis of skin fibrosis in this case could have been the direct and/or indirect consequence of the coexisting Th2-type immune response. PMID- 19300284 TI - Unmet patient needs in systemic sclerosis. AB - OBJECTIVE: Assessment of systemic sclerosis patients has not directly addressed functioning from the patient's perspective. With this study, we aim to gain our patient's point of view by using a questionnaire to describe their unmet needs and understanding what demographic parameters influence these. METHODS: A computer randomization program selected 50 patients, from 242 systemic sclerosis patients actively followed at our rheumatology clinic, to receive a survey about unmet needs. Twenty-five patients responded to the survey. Of 81 questions, 9 provided demographic data, whereas 72 questions addressed physical, daily living, psychologic, spiritual, existential, health services, health information, social support, and employment issues. A 4-point scale from no need to high need was used to rate all questions. Significant need was considered any issue for which more than 50% of patients reported a high need. The Fisher exact test was used to compare different demographic variables to unmet patient needs. RESULTS: The psychologic/spiritual/existential category had 9 questions reaching significance, the health services category had 5 significant questions, the physical category had 4 significant questions. Patients who had not attended college were more likely to have higher needs than patients who completed a college degree. Unmarried patients reported higher needs in 8 measures as compared with married patients, and patients in rural areas had higher needs in social support needs. CONCLUSIONS: The greatest prevalence of unmet needs in scleroderma patients were in the psychologic/spiritual/existential domain, such as being unable to do things they used to do, fear that the disease will worsen, anxiety and stress, feeling down or depressed, fears of physical disability, uncertainty about the future, change in appearance, keeping a positive outlook, and feeling in control. Significant differences were observed in unmet needs based on education, marital status, location, knowledge of disease, and age. Understanding each patient's specific unmet needs either through direct questioning or by the use of a questionnaire such as the one used for this study can help clinicians to give better care to each of our patients. PMID- 19300285 TI - Tuberculosis of the sacroiliac joint in a patient with systemic lupus erythematosus. PMID- 19300286 TI - Prevalence of thyroid dysfunction in systemic lupus erythematosus. AB - OBJECTIVE: To report the prevalence and clinical relevance of autoimmune thyroid disease and thyroid antibodies in 524 patients with SLE. METHODS: The medical charts of SLE patients followed in our rheumatology unit were reviewed to determine the prevalence and clinical associations of autoimmune thyroid disease. These findings were compared with the prevalence of autoimmune thyroid disease in 50 female adults. Chi(2) tests and Fisher exact tests were used in the comparison of the groups for the categorical variables and Mann-Whitney U test for the continuous variables. Spearman rank correlation was used to identify an association between thyroid symptoms and disease activity. RESULTS: Symptomatic autoimmune thyroid disease was observed in 32 of 524 (6.1%) SLE patients and in 1 of 50 controls (P > 0.05), predominantly hypothyroidism (28 SLE patients vs. in 1 control). Subclinical thyroid disease was identified in 60 (11.5%) and positive thyroid autoantibodies in the absence of thyroid disease in 89 of 524 (17%) SLE patients. Thyroid autoantibodies preceded the occurrence of clinical autoimmune thyroid disease in 70% of SLE patients. Sjogren syndrome (P = 0.001) and positive rheumatoid factor (P = 0.04) were more frequently observed in SLE patients with autoimmune thyroid disease when compared with SLE patients without autoimmune thyroid disease. Disease activity of the SLE was correlated with the presence of symptoms of hyperthyroidism (r = 0.4; P = 0.004). CONCLUSION: Our patients with SLE had a high prevalence of symptomatic and significantly more subclinical hypothyroidism and positive thyroid autoantibodies. Thyroid autoantibodies may precede the appearance of clinical autoimmune disease. Sjogren syndrome and positive rheumatoid factors were more frequently observed in SLE patients with autoimmune thyroid disease. We believe that, since symptoms of SLE and thyroid disease can be similar, that SLE patients should routinely been investigated for autoimmune thyroid disease. PMID- 19300287 TI - Osteonecrosis and antiphospholipid syndrome. AB - Osteonecrosis (ON), or avascular necrosis of bone, has been related to decreased blood flow to the bone. Many local and systemic factors have been implicated in the pathogenesis of ON, involving corticosteroid therapy, systemic lupus erythematosus (SLE), hemoglobinopathies, alcohol abuse, Caisson disease, Gaucher disease, and hypercoagulability states. We describe the case of a previously healthy young male with no history of corticosteroid therapy, who developed ON initially on the femoral head, and later on the humeral head with high levels of anticardiolipin antibodies (aCL), beta-2-glycoprotein 1 antibodies and positive lupus anticoagulant. The association between primary antiphospholipid syndrome (PAPS) and ON is controversial and few cases without other risk factors have been described. A review of ON pathogenesis and its relation with thrombotic microangiopathy because of PAPS is presented. PMID- 19300288 TI - Acrocyanosis from phenazopyridine-induced sulfhemoglobinemia mistaken for Raynaud phenomenon. AB - Rheumatologists are often asked to evaluate patients with Raynaud phenomenon. Occasionally, an alternate explanation is revealed such as acrocyanosis. Methemoglobinemia and sulfhemoglobinemia are rare causes of cyanosis that can be medication-induced. Both are known complications of therapy with phenazopyridine. We report an unusual case of a 45-year-old woman in whom sulfhemoglobinemia from chronic therapy with phenazopyridine was misdiagnosed as due to Raynaud phenomenon and limited scleroderma. This case illustrates the importance of taking into account medication-related adverse events when evaluating patients with Raynaud-like phenomenon. PMID- 19300289 TI - Doctor-patient interactions in Mexican patients with rheumatic disease. AB - OBJECTIVE: To evaluate the characteristics of physician-patient communications in Mexican patients with rheumatic diseases. METHODS: We prospectively recruited 200 consecutive patients with chronic rheumatic diseases from a rheumatology clinic affiliated with a secondary care hospital in Mexico. All participants completed face-to-face interviews and surveys to determine their perceptions of the medical interaction with the physician at the clinic. Patients were assessed immediately before and after their visits. The Perceptions of Involvement in Care Scale was used to examine 3 factors: the doctor's facilitation of involvement, the level of information exchange, and the patient's participation in decision making. The Medical Outcomes Study scale was used to evaluate the patient satisfaction with the visit. A Patients' Trust in Physicians instrument was also administered. RESULTS: No differences in communication, satisfaction, or trust were observed across diseases. Most patients indicated they wanted to play a passive role, with the physicians making the decisions. However, concordance between preferred and actual roles was small (weighted kappa = 0.3), suggesting that not all patients played their preferred role. CONCLUSION: This is the first study to evaluate doctor-patient communication in Mexican patients with rheumatic diseases. In general, patients preferred to play a passive role in their medical interaction. In addition, they often did not attain their preferred communication style when interacting with their physicians. The effect of these findings in subsequent health outcomes is unknown but deserves further investigation. PMID- 19300290 TI - Evaluation of risk factors that contribute to high prevalence of premature atherosclerosis in Chinese premenopausal systemic lupus erythematosus patients. AB - OBJECTIVE: To evaluate the prevalence of atherosclerosis in Chinese premenopausal women with systemic lupus erythematosus (SLE) and study possible associations between traditional and nontraditional risk factors with premature atherosclerosis. METHODS: We evaluated 111 premenopausal women with SLE and 40 healthy controls without clinical cardiovascular disease. B-mode ultrasound was used to measure carotid plaque and intima-media wall thickness (IMT). The frequency of risk factors for atherosclerosis in patients and controls was compared, and the relationship between the patients' clinical characteristics and carotid plaque was examined. At the same time, we used B-mode ultrasound to measure flow-mediated dilation (FMD) and nitroglycerin-mediated dilation (NMD) in the brachial artery to assess for difference in endothelial function between SLE patients and controls. RESULTS: Carotid plaque was more frequent in patients with lupus (16 of 111 patients) than in control subjects (0 of 40 subjects) (P = 0.007). The mean IMT was significantly higher in patients than in controls. Compared with controls, SLE patients were found to have a significantly higher prevalence of hypertension (P = 0.001), hypercholesterolemia (P = 0.022), and hypertriglyceridemia (P < 0.001). As compared with patients without plaque, patients with plaque were significantly older, had longer disease duration, higher body mass index, raised blood pressure, shorter prothrombin time, raised C reactive protein, higher Systemic Lupus International Collaborating Clinics damage index score, higher cumulative prednisone dose, used less hydroxychloroquine, had higher mean IMT, lower FMD, and NMD. In logistic regression analysis, older age, higher body mass index, and higher Systemic Lupus International Collaborating Clinics damage index score were independently related to the presence of plaque. Using multiple regression analysis, we found SLE (P = 0.003) to be significantly associated with impaired FMD. CONCLUSION: In our Chinese SLE group, patients presented a higher prevalence of carotid atherosclerosis plaque than healthy controls. SLE patients have significant endothelial dysfunction. We found that risk factors identified in other SLE populations were associated with atherosclerosis in our Chinese group. PMID- 19300291 TI - Electroconvulsive therapy in the setting of implantable cardioverter defibrillators. AB - Electroconvulsive therapy has potent cardiovascular effects, which may pose a challenge in treating patients with preexisting cardiac disease. Although it is well studied in the treatment of refractory depression, there are still pitfalls to overcome when treating those with comorbid cardiovascular disease. The synergistic effects of depression and cardiovascular disease in affecting the morbidity and mortality of patients make treatment a significant issue. The authors discuss 3 elderly patients treated for major depression, all of whom previously had implantable cardioverter-defibrillators placed. All 3 patients were effectively treated, with no major adverse effects. The cases illustrate how taking the appropriate preventative measures allow even the most medically challenging patient to undergo effective treatment and highlight the debate surrounding the intraoperative management of implantable cardioverter defibrillator devices. PMID- 19300292 TI - Neurophysiological characterization of high-dose magnetic seizure therapy: comparisons with electroconvulsive shock and cognitive outcomes. AB - Magnetic seizure therapy (MST) is under development as a means of improving cognitive outcomes with convulsive therapy through achieving better control over therapeutic seizure induction than is possible with conventional electroconvulsive therapy. In this investigation, we present the first neurophysiological characterization of high-dose MST (HD-MST, 6x seizure threshold) to see if a higher dose that is often used in human trials retains differential expression relative to electroconvulsive shock (ECS) and to explore the relationship between seizure expression and cognitive outcomes. To this end, rhesus monkeys received 4 weeks of daily treatment with ECS, HD-MST, and anesthesia-alone sham in counterbalanced order, with an interposed recovery period. Two channels of electroencephalogram were recorded during and immediately after the ictal period. Electroencephalogram power within delta, theta, alpha, and beta frequency bands was calculated. Electroconvulsive shock showed significantly more ictal power in all frequency bands than HD-MST (P < 0.01). Electroencephalogram power during the postictal period was significantly different among conditions only for the delta band. Higher ictal expression with ECS was associated with slowed completion time for an orientation task given immediately after the treatments. Our results support earlier findings demonstrating that MST- and ECS-induced seizures elicit differential patterns of ictal expression, consistent with the relatively more superficial stimulation achieved via magnetic induction in comparison with conventional electroconvulsive therapy. These results demonstrate that MST, even at high dose, results in seizures that differ neurophysiologically from ECS. It further suggests that some of the differences in ictal expression may relate to the improved cognitive outcomes seen with MST. PMID- 19300294 TI - The development of semantic priming effect in childhood: an event-related potential study. AB - To investigate the development of semantic priming effect in school children, we examined the event-related potentials in 120 children from 7 to 12 years when they were instructed to perform a lexical decision task. The well-known N400 component and a late positive component showed parabolic changes with the age growing. In addition, the lateralization of the brain potentials indicated the early stage in elementary school was the curial time for development of linguistic neural networks. This finding first revealed the detailed changes of semantic priming effect in childhood. PMID- 19300295 TI - Differences in early-phase tracer distribution in lung between 11C-acetate and 13N-ammonia. AB - AIM: N-ammonia and C-acetate are well-known tracers useful for the measurement of myocardial blood flow. Some studies reported that the measurement of C-acetate showed better statistical quality than that with N-ammonia. To evaluate this difference, we focused on the early-phase tracer kinetics in blood and lung. The tracer kinetics in the myocardium, arterial blood, venous blood, and lung were compared between these two tracers. PATIENTS AND METHODS: Twelve patients were studied. Dynamic positron emission tomography was performed with C-acetate and N ammonia within a 1-week period. The time activity curves (TACs) of the myocardium, arterial blood, and lung were calculated. The TACs normalized to the peak activity of right ventricle blood (normalized TAC) were also generated. RESULTS: The area under the curve of each TAC was calculated. Lung activity tended to be higher when assessed with N-ammonia. The area under the curve of the lung was larger with N-ammonia than with C-acetate both with the original TAC (478+/-232 vs. 248+/-87 MBq s/ml, P<0.001) and the normalized TAC (47.1+/-17.9 vs. 22.8+/-6.9 ratio x s, P<0.001). CONCLUSION: N-ammonia and C-acetate showed strikingly different early-phase tracer distribution in the lung. This difference may affect myocardial blood flow measurement, which largely depends on the early phase image quality. PMID- 19300296 TI - Labyrinthitis ossificans. PMID- 19300297 TI - Electrically evoked auditory brainstem responses in adults and children: effects of lateral to medial placement of the nucleus 24 contour electrode array. AB - OBJECTIVE: Previous electrophysiologic studies of electrode placement within the scala tympani in both animals and humans have shown effects on neural responses to electrical stimulation. The specific effects, however, may be dependent on electrode design, the location of an electrode along the array, and the method of perimodiolar positioning. The present study compares the effects of lateral and medial positioning of the Nucleus Contour electrode array on electrophysiologic responses in adult and pediatric subjects. STUDY DESIGN: Prospective clinical study. SETTING: Comprehensive Cochlear Implant Program/Tertiary Referral Center. PATIENTS: Subjects were adults (n = 15) and children (n = 20) who were consecutively implanted at our center with the Nucleus 24 Contour device. INTERVENTION: Rehabilitative. MAIN OUTCOME MEASURES: Intraoperative electrically evoked auditory brainstem responses (EABRs) were recorded within subjects for lateral and medial placement of the electrode array. Effects of electrode placement on EABR Wave V threshold and suprathreshold amplitude were measured. RESULTS: Group analyses showed significant decreases in EABR threshold and significant increases in EABR amplitude across all electrodes with medial electrode placement. The effects differed across electrode locations for the adult and pediatric subjects. No significant changes in Wave V input/output function slope were found. CONCLUSION: Medial electrode placement from stylet removal with the Nucleus 24 Contour array results in an increased neural response compared with the lateral condition as demonstrated by lower threshold and larger suprathreshold amplitude of the EABR. Possible clinical implications of these findings are lower psychophysical threshold and comfortable levels with medial cochlear electrode position. PMID- 19300298 TI - Morphology of the malleus handle and the comparison of different prostheses for malleostapedotomy. AB - OBJECTIVES: To assess malleus morphology to characterize optimal sites for prosthesis attachment in malleostapedotomy and to examine attachment, position, and depth of penetration in the vestibule of the 3 most used prostheses. METHODS: Ten mallei were processed for histologic examination with the light microscope. Using digitized video images of the histologic slices, the diameters, circumference, and shape of the specimen were determined. Implantation of 3 prostheses (Gyrus Nitinol piston, Storz titanium stapes piston, and Kurz malleovestibulopexy piston), was performed in 3 temporal bones for a total of 27 implantations. RESULTS: The cross-section of the malleus just distal to the lateral process shows an inclined oval shape with a mean minimum diameter of 0.84 +/- 0.10 mm, a mean maximum diameter of 1.02 +/- 0.23 mm, and a mean circumference of 3.23 +/- 0.49 mm. The quality of attachment to the malleus, the position of the prosthesis piston, and the depth of penetration were reliable for the Storz titanium stapes piston, satisfactory though variable for the Gyrus Nitinol piston and poor for the Kurz malleovestibulopexy piston as judged by contact with the malleus surface and predictability of insertion depth into the vestibule. CONCLUSION: The oval and inferoanteriorly inclined shape of the malleus distal to the lateral process requires the use of a prosthesis capable of molding itself to its surface for reliable attachment. To achieve the correct perpendicular position of the piston as it relates to the stapedotomy opening, individualized adaptation of the prosthesis shaft and loop to the anterior position of the malleus should be made in situ. PMID- 19300299 TI - Vestibular evoked myogenic potentials in normal mice and Phex mice with spontaneous endolymphatic hydrops. AB - OBJECTIVE AND BACKGROUND: Vestibular evoked myogenic potentials (VEMPs) have been recorded from the neck musculature and the cervical spinal cord in humans and a limited number of laboratory animals in response to loud sound. However, the mouse VEMP has yet to be described. Evaluation of the sacculocollic pathway via VEMPs in mice can set the stage for future evaluations of mutant mice that now play an important role in research regarding human auditory and vestibular dysfunction. MATERIALS AND METHODS: Sound-evoked potentials were recorded from the neck extensor muscles and the cervical spinal cord in normal adult mice and in circling Phex(Hyp-Duk/y) mice with known vestibular abnormalities, including endolymphatic hydrops (ELH). RESULTS: Biphasic potentials were recorded from all normal animals. The mean threshold of the VEMP response in normal adult mice was 60 dB normal hearing level with a mean peak latency of 6.25 +/- 0.46 and 7.95 +/- 0.42 milliseconds for p1 and n1 peaks, respectively. At the maximum sound intensity used (100 dB normal hearing level), 4 of 5 Phex mice did not exhibit VEMP responses, and 1 showed an elevated threshold, but normal response, with regard to peak latency and amplitude. The histologic findings in all of these Phex mice were consistent with distended membranous labyrinth, displaced Reissner membrane, ganglion cell loss, and ELH. CONCLUSION: This is the first report of VEMP recordings in mice and the first report of abnormal VEMPs in a mouse model with ELH. The characteristics of these potentials such as higher response threshold in comparison to auditory brainstem response, myogenic nature of the response, and latency correlation with the cervical recording (accessory nerve nucleus) were similar to those of VEMPs in humans, guinea pigs, cats, and rats, suggesting that the mouse may be used as an animal model in the study of VEMPs. The simplicity and reliability of these recordings make the VEMP a uniquely informative test for assessing vestibular function, and these results suggest that they may be informative in mice with various mutations. However, further investigation is necessary. PMID- 19300300 TI - Cochlear and petrous carotid canal erosion secondary to cholesteatoma. PMID- 19300301 TI - Functional and molecular expression of epithelial sodium channels in cultured human endolymphatic sac epithelial cells. AB - HYPOTHESIS: Epithelial sodium channels are expressed in cultured human endolymphatic sac (ES) epithelial (HESE) cells and epithelial sodium channel (ENaC) expression is suppressed by interleukin 1beta. BACKGROUND: The ES is part of the membranous labyrinth in the inner ear that plays an important role in maintaining homeostasis of the endolymphatic fluid system. However, the exact mechanism of fluid volume regulation is not yet known. METHODS: The ES specimens were harvested during acoustic neuroma surgery (n = 13) using the translabyrinthine approach and were subcultured with high-epidermal growth factor (25 ng/mL) media. RESULTS: The serially passaged HESE cells differentiated into a monolayer of confluent cells and some of the cultured cells had features of mitochondria-rich cells. Reverse transcription-polymerase chain reaction revealed that ENaC subunits are expressed in the cultured HESE cells. We also confirmed the presence of an ENaC-dependent short-circuit current in the cultured HESE cells. Interestingly, ENaC mRNA expression and ENaC-dependent current decreased after treatment with interleukin 1beta (10 nmol/L for 24 h). CONCLUSION: These findings suggest that ENaC plays an important role in fluid absorption in the human ES and that its function may be altered during inflammatory conditions. PMID- 19300302 TI - Mandibular condyle and tympanic plate fracture causing external auditory canal stenosis. PMID- 19300307 TI - "I am not afraid of dying. I just don't want to be there when it happens". PMID- 19300303 TI - Genetic and environmental influences on the ratio of 3'hydroxycotinine to cotinine in plasma and urine. AB - OBJECTIVES: The ratio of trans-3'hydroxycotinine/cotinine (3HC/COT) is a marker of CYP2A6 activity, an important determinant of nicotine metabolism. This analysis sought to conduct a combined genetic epidemiologic and pharmacogenetic investigation of the 3HC/COT ratio in plasma and urine. METHODS: One hundred and thirty-nine twin pairs [110 monozygotic and 29 dizygotic] underwent a 30-min infusion of stable isotope-labelled nicotine and its major metabolite, cotinine, followed by an 8-h in-hospital stay. Blood and urine samples were taken at regular intervals for analysis of nicotine, cotinine, and metabolites. DNA was genotyped to confirm zygosity and for variation in the gene for the primary nicotine metabolic enzyme, CYP2A6 (variants genotyped: *1B, *1 x 2, *2, *4, *9, *12). Univariate biometric analyses quantified genetic and environmental influences on each measure in the presence and absence of covariates, including measured CYP2A6 genotype. RESULTS: There was a substantial amount of variation in the free 3HC/COT ratio in plasma (6 h postinfusion) attributable to additive genetic influences (67.4%, 95% confidence interval=55.9-76.2%). The heritability estimate was reduced to 61.0 and 49.4%, respectively, after taking into account the effect of covariates and CYP2A6 genotype. In urine (collected over 8 h), the estimated amount of variation in the 3HC/COT ratio attributable to additive genetic influences was smaller (47.2%, 95% confidence interval=0-67.2%) and decreased to 44.6 and 42.0% after accounting for covariates and genotype. CONCLUSION: Additive genetic factors are prominent in determining variation in plasma 3HC/COT but less so in determining variation in urine 3HC/COT. PMID- 19300308 TI - The reporting rate of euthanasia and physician-assisted suicide: a study of the trends. AB - OBJECTIVES: To study trends in reporting rates of euthanasia from 1990 to 2005 in relation to whether recommended or nonrecommended drugs were used, and the most important differences between reported and unreported cases in 2005. RESEARCH DESIGN: Questionnaires were sent to a sample of 6860 physicians who had reported a death in 2005 (response 78%). Previously, 3 similar studies were done at 5-year intervals. The total number of euthanasia and physician-assisted suicide cases was estimated using a "gold standard" definition: death was-according to the physician-the result of the use of drugs at the explicit request of the patient with the explicit goal of hastening death (denominator). The Euthanasia Review Committees provided the number of reported cases (numerator). RESULTS: The reporting rate of euthanasia and physician-assisted suicide increased from 18% in 1990, 41% in 1995, and 54% in 2001 to 80% in 2005. The reporting rate in the subgroup of euthanasia with recommended drugs (barbiturates and muscle relaxants) was 73% in 1995, 71% in 2001, and 99% in 2005. The reporting rate of euthanasia with nonrecommended drugs (eg, opioids) was below 3% in 1995, 2001, and 2005. Unreported euthanasia differed also from reported euthanasia in the fact that physicians less often labeled their act as euthanasia. CONCLUSIONS: Euthanasia with nonrecommended drugs is almost never reported. The total reporting rate increased because of an increase in the use of recommended drugs for euthanasia between 1995 and 2001, and an increase in the reporting rate for euthanasia with recommended drugs between 2001 and 2005. PMID- 19300309 TI - A population-based study of age inequalities in access to palliative care among cancer patients. AB - BACKGROUND: Inequalities in access to palliative care programs (PCP) by age have been shown to exist in Canada and elsewhere. Few studies have been able to provide greater insight by simultaneously adjusting for multiple demographic, health service, and socio-cultural indicators. OBJECTIVE: To re-examine the relationship between age and registration to specialized community-based PCP programs among cancer patients and identify the multiple indicators contributing to these inequalities. METHODS: This retrospective, population-based study was a secondary data analysis of linked individual level information extracted from 6 administrative health databases and contextual (neighborhood level) data from provincial and census information. Subjects included all adults who died due to cancer between 1998 and 2003 living within 2 District Health Authorities in the province of Nova Scotia, Canada. The relationship between registration in a PCP and age was examined using hierarchical nonlinear regression modeling techniques. Identification of potential patient and ecologic contributing indicators was guided by Andersen's conceptual model of health service utilization. RESULTS: Overall, 66% of 7511 subjects were registered with a PCP. Older subjects were significantly less likely than those <65 years of age to be registered with a PCP, in particular those aged 85 years and older (adjusted odds ratio: 0.4; 95% confidence interval: 0.3-0.5). Distance to the closest cancer center had a major impact on registration. CONCLUSIONS: Age continues to be a significant predictor of PCP registration in Nova Scotia even after controlling for the confounding effects of many new demographic, health service, and ecologic indicators. PMID- 19300310 TI - Physician gender affects how physician nonverbal behavior is related to patient satisfaction. AB - BACKGROUND: Physician and patient gender both influence medical communication. Nonverbal behavior is generally under-researched in the medical encounter but plays an important role for patient outcomes such as satisfaction. OBJECTIVE: This article aims at identifying how specific physician nonverbal behaviors predict analogue patient satisfaction depending on physician and patient gender. RESEARCH DESIGN: Eleven physicians in a real medical encounter were videotaped and analogue patients indicated their satisfaction with each physician while viewing the videotapes. SUBJECTS: One hundred sixty-three university students participated (analogue patients). MEASURES: From the videotapes, 17 physician nonverbal behaviors (related to face, body, voice/speech), 2 physician appearance cues, 2 characteristics of the examination room, and 1 patient behavior were coded. For each analogue patient, the correlation between each of these coded characteristics and the patient's satisfaction was calculated, across all physicians and across male and female physicians separately. RESULTS: There was no main effect for patient gender but most coded characteristics showed different relations to patient satisfaction according to physician gender. Analogue patients were most satisfied with female physicians who behaved in line with the female gender role (eg, more gazing, more forward lean, softer voice) while still stressing their professionalism (laboratory coat, medical-looking examination room). For male physicians, satisfaction was high for a broader range of behaviors, partly related to their gender role (eg, louder voice, more distance to patient). CONCLUSIONS: To be satisfied, patients expect female and male physicians to show different patterns of nonverbal behavior. Awareness of these gender-specific expectations should be taken into account in medical training. PMID- 19300311 TI - Lapses in Medicaid coverage: impact on cost and utilization among individuals with diabetes enrolled in Medicaid. AB - BACKGROUND: Gaps in Medicaid coverage can result in inadequate access to care. This can be particularly detrimental to those with a chronic disease such as diabetes. OBJECTIVE: To assess whether a lapse in Medicaid coverage is associated with an increase in expenditures, and acute care utilization upon reenrollment among beneficiaries with diabetes. RESEARCH DESIGN: Using multivariate regression analyses, we compared pre- versus post-expenditures and utilization among 2102 individuals with diabetes who had experienced at least one 1-month lapse in their Medicaid coverage. MEASURES: Dependent variables were the number of inpatient episodes, total length of stay, total number of emergency room visits, total expenditure, and pharmaceutical expenditures. These were aggregated over 3-month spans that either immediately preceded or immediately followed a lapse in coverage. Key predictor variables included a variable that identified the span as occurring pre-lapse or post-lapse in coverage, and a continuous variable identifying the length of the lapse. Predicted expenditure and utilization were calculated. RESULTS: Overall total program expenditures were higher for post lapse periods compared with pre-lapse periods. Total expenditures were estimated to increase by $239 per member per month for the 3-month period. The likelihood of having any expenditure was actually lower in the post-lapse period. However inpatient and emergency room use was higher. CONCLUSIONS: The results from this study suggest that interruptions in Medicaid coverage are associated with overall greater program expenditures in the post-lapse periods. However, this increase in expenditures seems to be driven by a subset of individuals whose greater use of inpatient and emergency room services increased overall program costs. PMID- 19300312 TI - Cost savings attributable to reductions in intensive care unit length of stay for mechanically ventilated patients. AB - OBJECTIVES: To estimate the actual cost savings that could be achieved through reductions in intensive care unit (ICU) length of stay and duration of mechanical ventilation by determining the short-run marginal variable cost of an ICU and ventilator day. RESEARCH DESIGN: Retrospective cohort study in a university affiliated teaching hospital. SUBJECTS: All patients receiving mechanical ventilation in the ICU for more than 48 hours (n = 1778) from July 1, 2005 to June 30, 2006. MEASURES: The hospital's administrative and cost databases were used to determine total costs, variable costs, and direct-variable costs for each patient on each individual ICU and hospital day. RESULTS: Direct-variable costs comprised 19.3% of total ICU costs and 18.4% of total hospital costs. Marginal direct-variable costs (the cost of each additional ICU day) were small compared with the average daily total cost ($649 to $839 vs. $1751, in US dollars). In survivors with ICU lengths of stay more than 3 days, the mean direct-variable cost of the last ICU day was $397, while the mean direct-variable cost of the first ward day was $279, for a mean cost difference of $118 (95% CI, $21-$190). Reducing ICU and hospital length of stay by 1 day in all survivors with ICU lengths of stay more than 3 days would result in an immediate cost savings of only 0.2% of all hospital expenditures for these patients. CONCLUSIONS: Marginal variable ICU costs are relatively small compared with average total costs and are only slightly greater than the cost of a ward day. PMID- 19300313 TI - Time to first dental visit after initially enrolling in Medicaid and S-SCHIP. AB - BACKGROUND: Accessing dental care is a significant problem for children in Medicaid and S-SCHIP (Separate State Children's Health Insurance Program). Previous studies have indicated that the design of the Medicaid or S-SCHIP dental program can have an impact on the ability to use services. OBJECTIVE: To evaluate the factors related to how quickly children had any dental visit and had a preventive dental visit after first enrolling in the Iowa Medicaid and S-SCHIP programs. The primary question was whether the structure of the dental plan was related to improved access to care. METHODS: Iowa Medicaid and S-SCHIP dental claims and enrollment files for FY 2001 through 2003 were used to identify children who were newly enrolled in the programs and their use of dental services. Factors related to the time to a child's first dental visit were analyzed using survival analytic techniques. RESULTS: After 6 months in the program, between 21% and 36% of children had received their first dental visit, depending on their dental plan. This increased from 39% to 56% after 1 year. Based on the survival analysis, earlier dental utilization was related to the type of plan in which the child was enrolled as well as the child's age, race, and urban/rural location. CONCLUSIONS: Children in the S-SCHIP 2 dental plan, which had an open provider network and paid dentists' full charges, were most likely to have had a dental visit sooner after enrollment. States looking for options to improve access to dental care in their Medicaid and S-SCHIP programs should consider contracting with dental plans with these features. PMID- 19300315 TI - Mental illness and hospitalization for ambulatory care sensitive medical conditions. AB - BACKGROUND: Hospitalization due to ambulatory care sensitive (ACS) medical conditions is widely used as an indicator of poor primary care access and effectiveness. It is unknown whether patients with mental disorders have higher ACS admission rate, compared with patients without mental disorders. OBJECTIVE: To compare the ACS admission pattern and its resultant hospital cost and length of stay (LOS) between medical patients with and without coexisting mental disorders. METHODS: Using New York State hospital discharge data for 2004, we conducted a retrospective cohort study on inpatient cases who were aged 20-64 years and hospitalized due to either ACS condition or non-ACS "marker" condition. Multivariate regression was used to estimate the relative odds of ACS admissions and the incremental resource use for mentally ill patients during ACS hospitalization. RESULTS: Inpatient cases with mental disorders (N = 38,514) were more likely than others (N = 116,798) to have ACS admission [adjusted odds ratio (AOR), 2.30; 95% confidence interval (CI), 2.17-2.43] relative to admission due to marker conditions. During ACS hospitalization, mentally ill cases showed an average incremental cost of $556 (95% CI, $340-$778), and an average incremental LOS of 0.7 days (95% CI, 0.6-0.8 days). The higher ACS admission rate and hospital resource consumption were most pronounced for those with major depression, other psychoses, and combined psychiatric and substance-abuse disorders. CONCLUSIONS: Patients with mental disorders experience higher risk of hospitalization due to ACS medical conditions than the general population. During an ACS hospitalization, patients with mental disorders have longer length of stay and higher hospital cost than other patients. PMID- 19300314 TI - Sexual orientation and testing for prostate and colorectal cancers among men in California. AB - BACKGROUND: Previous quantitative studies have not compared the use of prostate and colorectal cancer (CRC) testing between gay/bisexual and heterosexual men. METHODS: We analyzed cross-sectional data on 19,410 men in the California Health Interview Survey. The percentage of respondents age 50 and over who received prostate and CRC tests was calculated across subgroups defined by self-reported sexual orientation, race/ethnicity, and a combined variable on sexual orientation and race/ethnicity. Multivariate regression analysis was used to identify variables on respondent characteristics that were independently associated with testing. RESULTS: In bivariate analyses, the percentage of gay/bisexual men receiving CRC tests was 6-10% greater than that of heterosexuals. There were no overall differences in prostate-specific antigen (PSA) test use between gay/bisexual and heterosexual men; however, use of these tests by gay/bisexual African Americans was 12-14% lower than that of heterosexual African Americans and 15-28% lower than that of gay/bisexual whites. In multivariate analyses, gay/bisexual men had greater odds of ever receiving CRC tests [odds ratio (OR) = 1.67; 95% confidence interval (CI) = 1.06-2.65], and lower odds of having an up to-date PSA test than did heterosexuals (OR = 0.61; 95% CI = 0.42-0.89). However, interactions between sexual orientation and living situation showed that gay/bisexual men who lived alone had greater odds of receiving PSA tests than did other men (OR = 1.93; 95% CI = 1.23-3.03). CONCLUSIONS: Sexual orientation is independently associated with cancer testing among men. Future work should investigate the differences in this association by race/ethnicity and living situation. PMID- 19300316 TI - Can a community evidence-based asthma care program improve clinical outcomes?: a longitudinal study. AB - RATIONALE: Asthma is associated with significant morbidity. Previous studies highlight significant variations in asthma management approaches within primary care settings where the adoption of published asthma guidelines is typically suboptimal. OBJECTIVE: To determine whether the implementation of an evidence based asthma care program in community primary care settings leads to improved clinical outcomes in asthma patients. METHODS, MEASUREMENTS, AND MAIN RESULTS: A community-based participatory research project was implemented at 8 primary care practices across Ontario, Canada, consisting of elements based on the Canadian Asthma Consensus Guidelines (asthma care map, program standards, management flow chart and action plan). A total of 1408 patients aged 2-55 years participated. Conditional logistic regression analyses were used to calculate the odds ratios (OR) comparing baseline to follow-up while adjusting for age, gender, socioeconomic status and other covariates. At 12-month follow-up, there were statistically significant reductions in self-reported asthma exacerbations from 77.8% to 54.5% [OR = 0.35; 95% confidence interval (CI): 0.28-0.43]; emergency room visits due to asthma from 9.9% to 5.5% (OR = 0.47; 95% CI: 0.32-0.62); school absenteeism in children from 19.9% to 10.2% (OR = 0.37; 95% CI: 0.25 0.54); productivity loss in adults from 12.0% to 10.3% (OR = 0.49; 95% CI: 0.34 0.71); uncontrolled daytime asthma symptoms from 62.4% to 41.4% (OR = 0.34; 95% CI: 0.27-0.42); and uncontrolled nighttime asthma symptoms from 46.4% to 25.4% (OR = 0.29; 95% CI: 0.23-0.37). CONCLUSIONS: Development and implementation of a community-based primary care asthma care program led to risk reductions in exacerbations, symptoms, urgent health service use and productivity loss related to asthma. PMID- 19300317 TI - Electronic health record components and the quality of care. AB - BACKGROUND: Electronic health records (EHRs) have been promoted as an important tool to improve quality of care. We examined the association between EHR components, a complete EHR, and the quality of care. METHODS: Using data from the 2005 National Ambulatory Medical Care Survey and the National Hospital Ambulatory Medical Care Survey, we conducted a cross-sectional analysis of all visits with an established primary care provider and examined the association between presence of EHR components and: (1) blood pressure control; and (2) receipt of appropriate therapy for chronic conditions. We examined similar associations for complete EHRs which we defined as one that includes physician and nursing notes, electronic reminder system, computerized prescription order entry, test results, and computerized test order entry. We constructed multivariate models to examine the association between EHR components and each outcome controlling for patient sociodemographic, health, physician practice, and geographic factors. RESULTS: We found no association between electronic physician notes and blood pressure control or receipt of appropriate therapies, with the exception of inhaled steroids among asthmatics (adjusted odds ratio 2.86; 95% confidence interval, 1.12-7.32). We found no association between electronic reminder systems and blood pressure control or receipt of appropriate therapies, with the exception of angiotensin converting enzyme inhibitors or angiotensin receptor blockers in patients with diabetes with hypertension (odds ratio 2.58; 95% confidence interval, 1.22-5.42). We found no association between electronic physician notes and any measure of quality. We found no relationship between having a complete EHR and any of the quality measures investigated. CONCLUSIONS: We found no consistent association between blood pressure control, management of chronic conditions, and specific EHR components. Future research focusing on how an EHR is implemented and used and how care is integrated through an EHR will improve our understanding of the impact of EHRs on the quality of care. PMID- 19300319 TI - Thoughts on medical liability costs and the future of health care. PMID- 19300320 TI - Decreased preterm births in an inner-city public hospital. AB - OBJECTIVE: To examine preterm births among African-American and Hispanic women who delivered at an inner-city public hospital in the context of contemporaneously increasing rates in the United States. METHODS: The rates of preterm birth, defined as birth before 37 weeks of gestation, for singleton neonates with birth weights of 500 g or higher delivered to women who had prenatal care at Parkland Hospital between 1988 and 2006 were compared with similar births in the United States from 1995 to 2002. Preterm birth rates also were compared for white, African-American, and Hispanic women as were disparity in these rates using white women as the referent. RESULTS: The Parkland Memorial Hospital cohort included 260,197 women, of whom 70% were Hispanic, 20% African American, and 8% white. The U.S. cohort included 29,366,816 women, of whom 61% were white, 19% Hispanic, and 14% African-American. Between 1995 and 2002, the rate of preterm birth in the United States increased from 9.4% to 10.1% (P<.001). Between 1988 and 2006, the rate of preterm births at Parkland decreased from 10.4% to 4.9% (P<.001). Moreover, the rates of preterm birth were significantly lower in Hispanic and African-American women who delivered at Parkland compared with the same ethnicity/race groups in the U.S. cohort. The decline in preterm births at Parkland Memorial Hospital coincided with increased prenatal care use. CONCLUSION: Preterm birth significantly decreased in minority women delivered at an inner-city public hospital. We hypothesize that the reduction in preterm births was the result of a public health care program specifically targeting minority pregnant women. PMID- 19300321 TI - Tocolytic therapy: a meta-analysis and decision analysis. AB - OBJECTIVE: To determine the optimal first-line tocolytic agent for treatment of premature labor. METHODS: We performed a quantitative analysis of randomized controlled trials of tocolysis, extracting data on maternal and neonatal outcomes, and pooling rates for each outcome across trials by treatment. Outcomes were delay of delivery for 48 hours, 7 days, and until 37 weeks; adverse effects causing discontinuation of therapy; absence of respiratory distress syndrome; and neonatal survival. We used weighted proportions from a random-effects meta analysis in a decision model to determine the optimal first-line tocolytic therapy. Sensitivity analysis was performed using the standard errors of the weighted proportions. RESULTS: Fifty-eight studies satisfied the inclusion criteria. A random-effects meta-analysis showed that all tocolytic agents were superior to placebo or control groups at delaying delivery both for at least 48 hours (53% for placebo compared with 75-93% for tocolytics) and 7 days (39% for placebo compared with 61-78% for tocolytics). No statistically significant differences were found for the other outcomes, including the neonatal outcomes of respiratory distress and neonatal survival. The decision model demonstrated that prostaglandin inhibitors provided the best combination of tolerance and delayed delivery. In a hypothetical cohort of 1,000 women receiving prostaglandin inhibitors, only 80 would deliver within 48 hours, compared with 182 for the next best treatment. CONCLUSION: Although all current tocolytic agents were superior to no treatment at delaying delivery for both 48 hours and 7 days, prostaglandin inhibitors were superior to the other agents and may be considered the optimal first-line agent before 32 weeks of gestation to delay delivery. PMID- 19300322 TI - Five-year experience of human papillomavirus DNA and Papanicolaou test cotesting. AB - OBJECTIVE: To estimate the 5-year age group-specific test positives for Pap tests and human papillomavirus (HPV) testing in a large, general screening population of women 30 and older. METHODS: Using data from Kaiser Permanente Northern California, a large health maintenance organization that introduced cotesting in 2003, we evaluated the cotesting results overall and by 5-year age groups. Women (n=580,289) who opted for and underwent cotesting (n cotests=812,598) between January 2003 and April 2008 were included in the analysis. Pap tests interpreted as atypical squamous cells of undetermined significance (ASC-US) or more severe were considered to be positive. Women were tested for carcinogenic HPV using an assay approved by the U.S. Food and Drug Administration. Binomial exact 95% confidence intervals (CIs) were calculated. RESULTS: Overall, 6.27% (95% CI 6.21 6.32%) of cotests were carcinogenic HPV positive, and only 3.99% (95% CI 3.94 4.03%) cotests had normal cytology and were carcinogenic HPV positive. By comparison, 5.18% (95% CI 5.13-5.23%) of cotests had ASC-US or more severe cytology, and 2.87% (95% CI 2.84-2.91%) of cotests had ASC-US or more severe cytology and were carcinogenic HPV negative. CONCLUSION: In a general screening population, concerns about excessive HPV test positives among women aged 30 years and older are not borne out. PMID- 19300323 TI - Risk factors for pelvic floor repair after hysterectomy. AB - OBJECTIVE: Having demonstrated that prior history of prolapse was a risk factor for pelvic floor repair procedures after hysterectomy, the objective of this study was to assess medical risk factors for pelvic floor repair after hysterectomy. METHODS: Using the Rochester Epidemiology Project database of 8,220 Olmsted County, Minnesota, women who had hysterectomy for benign indications from 1965-2002, we conducted a nested case-control study in 144 pairs, comparing women who underwent pelvic floor repair after hysterectomy (case group) with women matched for known risk factors (ie, age, pelvic floor disorders at baseline, year and type of hysterectomy, and pelvic floor repair during hysterectomy) (control group). RESULTS: The median duration between hysterectomy and pelvic floor repair was 13 years. Chronic pulmonary disease (odds ratio [OR] 14.3, 95% confidence interval [CI] 1.2-178), but not obstetric history, obesity, indication for hysterectomy, or chronic constipation, was associated with an increased risk of pelvic floor repair after hysterectomy. Between the hysterectomy and subsequent pelvic floor repair, overall pelvic organ prolapse severity changed by one grade or less in 54 case patients (38%, group A) but increased by two or more grades in 72 case patients (50%, group B). In group A, but not group B, uterine prolapse (OR 25, 95% CI 2.1-300) and chronic pulmonary disease (OR 22, 95% CI 1.5-328) at baseline remained risk factors for pelvic floor repair after hysterectomy. CONCLUSION: In this matched case-control study, chronic pulmonary disease was the only risk factor for pelvic floor repair after hysterectomy for benign indications, underscoring the need to address pulmonary status before surgery. PMID- 19300324 TI - Symptomatic pelvic organ prolapse at midlife, quality of life, and risk factors. AB - OBJECTIVE: To estimate quality of life, prevalence, and risk factors associated with symptomatic pelvic organ prolapse (POP) among middle-aged women. METHODS: This was a cross-sectional study. A questionnaire was mailed to 3,114 women aged 50-61 years in the GAZEL cohort; 2,640 (85%) returned it. Symptomatic POP was defined by feeling a bulge from the vagina (sometimes, often, or all the time compared with never or rarely). Quality of life was determined with the Nottingham Health Profile questionnaire. Multiple linear regression was used to examine the association between frequency of POP symptoms and the quality-of-life score. Logistic regression was used to estimate the effect of risk factors on past or present symptomatic POP (current symptoms or previous surgery for POP). RESULTS: The prevalence of symptomatic POP was 3.6% (96 of 2,640) and that of surgery for POP was 2.7% (70 of 2,640). Pelvic organ prolapse symptoms were associated with difficulty defecating, lower abdominal pain, and difficulty voiding. The frequency of POP symptoms was associated with a poorer quality-of life score in each Nottingham Health Profile domain (physical mobility, pain, emotional reaction, social isolation, energy, and sleep). Even when we took general characteristics, medical history, and lifestyle associated with quality of life into account, the global Nottingham Health Profile score was significantly impaired by POP symptoms. Factors significantly associated with past or present symptomatic POP were high body mass index and the number of vaginal deliveries. CONCLUSION: In our population of women in their 50s, POP symptoms are associated with impaired quality of life, and the number of vaginal deliveries is a risk factor for past or present symptomatic POP. PMID- 19300325 TI - Effect of transobturator tape on overactive bladder symptoms and urge urinary incontinence in women with mixed urinary incontinence. AB - OBJECTIVE: To estimate changes in overactive bladder (OAB) symptoms and urge urinary incontinence (UUI) in patients undergoing the transobturator tape procedure for urinary stress and mixed incontinence. METHODS: Telephone interviews were conducted using the International Consultation on Incontinence Female Lower Urinary Tract Symptoms questionnaire, the International Consultation on Incontinence-Overactive Bladder (ICIQ-OAB) questionnaire, and the Verbal Analogue Satisfaction (VeAS) scale. Preoperative OAB scores were compared with postoperative scores in women with stress incontinence only (group 1), mixed incontinence with predominant stress leakage (group 2), and mixed incontinence with predominant urge (group 3). Case notes were reviewed for preoperative assessment and complications. RESULTS: At median follow-up of 13 months, significant improvement was noted in ICIQ-OAB scores, from a median of 10 (1-15) preoperatively to a median of 3 (0-11) postoperatively (P<.001). Overall, UUI was cured in 19 of 44 (43%) patients, improved in a further 16 (36%), and was persistent in only 9 (21%). In group 2 (stress predominant), UUI was cured in 10 of 23 (43.5%) patients, improved in 10 (43.5%), and persistent in three (13%). In group 3 (urge predominant), UUI was cured in 9 of 21 (43%) patients, improved in six (28.5%), and persistent in six (28.5%). Postoperative lower urinary tract symptom scores were low in all three groups (median 4/48 [0-18]). Stress incontinence was cured in 77%, improved in a further 19%, and unchanged in 4%. Median VeAS score was 9 (2-10); 21% (11/52) of participants had low satisfaction scores (less than 8) owing to persistent urge and slow voiding. CONCLUSION: Marked resolution or improvement (79%) in urge incontinence after the transobturator tape procedure was noted, and no cases of de novo urge incontinence were identified. PMID- 19300326 TI - Time from decision to incision for cesarean deliveries at a community hospital. AB - OBJECTIVE: To compare decision-to-incision times of cesarean deliveries for various indications, estimate the percentage of cesarean deliveries initiated within 30 minutes (defined as "efficient"), and assess the effect of performance improvement projects over time. METHODS: In 2004 (n=291) and 2006 (n=574) cesarean deliveries were prospectively tracked for decision-to-incision time. Information collected included indication, urgency, and reason for delay. Performance improvement addressed surgeon, anesthesiologist, operating room availability, nursing availability, and patient-related delays. RESULTS: Unscheduled cesarean delivery efficiency improved from 36% in 2004 to 59% in 2006 (P<.001). Emergency cases improved from 67% to 84% (P=.07). The "Other" category was most improved over time, from 18% efficient deliveries in 2004 to 49% in 2006 (P<.001). For patients with failure to progress, efficiency improved from 33% in 2004 to 54% in 2006 (P<.001). For patients with nonreassuring fetal heart rate tracings, efficiency improved from 56% in 2004 to 76% in 2006 (P=.02). The efficiency rate for cesarean deliveries varied significantly by indication in both study years (2004, P<.001; 2006, P<.001). CONCLUSION: Cesarean delivery efficiency varied by indication, but a data-driven hospital-based initiative successfully improved efficiency over time. Indication-based standards that address an institution's staff and structural capacity are needed to evaluate time from decision to incision. PMID- 19300327 TI - Prevalence of uterine leiomyomas in the first trimester of pregnancy: an ultrasound-screening study. AB - OBJECTIVE: To estimate the proportion of pregnant women with one or more leiomyomas detected by research-quality ultrasound screening in the first trimester, to describe the size and location of leiomyomas identified, and to report variation in prevalence by race/ethnicity. METHODS: Within an ongoing prospective cohort, we conducted 4,271 first-trimester or postmiscarriage ultrasound examinations. Sonographers measured each leiomyoma three separate times, recording the maximum diameter in three perpendicular planes each time. Sonographers and investigators classified type and location. RESULTS: Among 458 women with one or more leiomyomas (prevalence 10.7%), we identified a total of 687 leiomyomas. The mean size of the largest leiomyoma was 2.3 cm (95% confidence interval [CI] 1.8-2.8). Mean gestational age at ultrasonography was 61+/-13 days from last menstrual period. Prevalence varied by race/ethnicity: 18% in African American women (95% CI 13-25), 8% in white women (95% CI 7-11), and 10% in Hispanic women (95% CI 5-19). The proportion of women with leiomyomas increased with age much more steeply for African-American women than for white women. CONCLUSION: Leiomyomas are common in pregnancy and occur more often among African American women. Given the limited research on effects of leiomyomas on reproductive outcomes, the degree to which race/ethnic disparities in prevalence of leiomyomas may contribute to disparities in events such as miscarriage and preterm birth warrants investigation. PMID- 19300328 TI - Predicting perinatal outcome through changes in umbilical artery Doppler studies after antenatal corticosteroids in the growth-restricted fetus. AB - OBJECTIVE: To investigate whether persistently absent umbilical artery end diastolic flow in the intrauterine growth-restricted fetus after betamethasone administration is associated with altered perinatal outcomes. METHODS: This is a retrospective cohort study of 92 pregnancies complicated by intrauterine growth restriction (IUGR) and absent end-diastolic flow in which antenatal betamethasone was given. Predefined maternal outcomes (maternal age, gestational age at diagnosis of absent end-diastolic flow, gestational age at delivery, preexisting medical conditions) and neonatal outcomes (including birth weight; perinatal mortality; duration of neonatal intensive care unit admission; requirement for intubation, assisted ventilation, inotropic support; duration of supplemental oxygen, assisted ventilation; respiratory distress syndrome, necrotizing enterocolitis, intraventricular hemorrhage) were analyzed. RESULTS: Betamethasone administration was associated with a transient return of end-diastolic umbilical artery flow in 58 pregnancies (63%) and persistent absent end-diastolic flow in 34 (37%). Persistent absent end-diastolic flow was seen more frequently in women with prepregnancy medical disorders (59% compared with 24%, P<.001). Neonates from the persistent absent end-diastolic flow subgroup were more likely to require assisted ventilation (93.1% compared with 73.5%, P=.03) and to have longer durations of assisted ventilation (median time 30 days compared with 4 days, P=.03) and supplemental oxygen (median time 45 days compared with 4 days, P=.04). CONCLUSION: Betamethasone administration is associated with a transient return of end-diastolic flow in two thirds of pregnancies complicated by IUGR and umbilical artery absent end-diastolic flow. Persistent absent end-diastolic flow in the umbilical artery after betamethasone administration may identify a subgroup of fetuses with IUGR at further heightened perinatal risk that, as neonates, are more likely to require assisted ventilation and a longer duration of ventilation and supplemental oxygen. PMID- 19300329 TI - Influence of maternal body mass index on the clinical estimation of fetal weight in term pregnancies. AB - OBJECTIVE: To estimate the effect of maternal body mass index (BMI) on clinical estimated fetal weight in term pregnancies. METHODS: We compared the clinical estimated fetal weight to the actual birth weight on 400 consecutive term pregnant patients admitted for delivery. We then evaluated whether the maternal BMI was significantly associated with the clinical estimated fetal weight accuracy in estimating birth weight. Patients with singleton pregnancies at 37-42 weeks of gestation were included. Patients with an ultrasound-estimated fetal weight within 3 weeks of admission were excluded. Estimated fetal weight accuracy was defined as the absolute error (absolute value of estimated fetal weight minus birth weight) and absolute percent error (absolute error divided by birth weight multiplied by 100). The primary outcome was an absolute percent error of less than 10% (ie, an estimated fetal weight within +/-10% of the birth weight). RESULTS: The proportion of clinical estimated fetal weights within +/-10% and within +/-20% of the birth weight significantly decreased with increasing BMI categories (chi for trend P=.040 and 0.020, respectively). Clinical estimated fetal weights obtained in women with BMI at or greater than 30 were significantly less likely to be within +/-10% of the birth weight when compared with women with a BMI less than 25 (66.4% compared with 82.5%, P=.011). Body mass index was significantly positively associated with the absolute error (P=.046) and the absolute percent error (P=.011), even after controlling for birth weight. CONCLUSION: Increased maternal BMI is significantly associated with decreased clinical estimated fetal weight accuracy. PMID- 19300330 TI - Antihypertensive therapy and central hemodynamics in women with hypertensive disorders in pregnancy. AB - OBJECTIVE: To estimate the changes in central hemodynamics features of pregnant women presenting with hypertensive disorders and to analyze the effects of standard antihypertensive treatment on maternal central hemodynamics. METHODS: Applanation tonometry was used to record the radial artery pulse waveform in 80 women presenting with preeclampsia or gestational hypertension and 80 normotensive controls matched for gestational age. In each case, an averaged aortic waveform was derived and analyzed to calculate augmentation pressure and augmentation index at heart rate 75 beats per minute (bpm). RESULTS: In women with preeclampsia and gestational hypertension, both augmentation pressure (P<.001 and P<.05, respectively) and augmentation index at heart rate 75 bpm (P<.001 and P<.001, respectively) were significantly higher than in controls. Augmentation pressure and augmentation index at heart rate 75 bpm were significantly higher in early- compared with late-onset preeclampsia (P<.001) and in severe compared with mild preeclampsia (P<.001). Antihypertensive therapy with alpha methyldopa resulted in a significant fall in both augmentation pressure and augmentation index at heart rate 75 bpm in preeclampsia (P<.001) but not in gestational hypertension. CONCLUSION: Arterial stiffness is increased in women with hypertensive disorders of pregnancy compared with normotensive controls. In preeclampsia, vascular stiffness was significantly improved by antihypertensive treatment with alpha methyldopa, but remained higher than in normotensive controls. PMID- 19300331 TI - Progestin therapy of complex endometrial hyperplasia with and without atypia. AB - OBJECTIVE: To assess the likelihood of histologic persistence/progression of complex hyperplasia and atypical hyperplasia among women treated with progestin compared with those not treated, with attention to type, dose, and duration. METHODS: This was a cohort study at an integrated health plan of women, ages 18 85 years, with complex or atypical hyperplasia on independent pathology review with a second endometrial specimen in the 2-6 months after the index diagnosis. Progestin therapy between index diagnosis and follow-up biopsy was determined from the pharmacy database. Medical record abstraction was performed. Relative risks (RRs), adjusted for age and body mass index, were calculated. RESULTS: Among 185 women, average age 55.9 years, follow-up 16.1 weeks, 115 had complex and 70 had atypical hyperplasia. Among women with complex hyperplasia, 28.4% of those treated with progestin and 30.0% of those not treated had persistence/progression (RR 1.20, 95% confidence interval [CI] 0.53-2.72). Among women with atypical hyperplasia, 26.9% of those treated with progestin and 66.7% of those not treated had persistence/progression (RR 0.39, 95% CI 0.21-0.70); there was a suggestion that use of at least a medium dose, or a duration of at least 3 months, was associated with a particularly low probability of persistence/progression. CONCLUSION: Although progestin treatment of women with atypical hyperplasia was associated with a substantial increase in the likelihood of regression of the lesion during the ensuing 2-6 months, persistence/progression was nonetheless present in more than one quarter of treated women. Regression of complex hyperplasia without atypia was common whether progestin had or had not been used. PMID- 19300332 TI - Rates of follow-up and repeat pregnancy in the 12 months after first-trimester induced abortion. AB - OBJECTIVE: To estimate the proportion of women who returned for a routine follow up visit after elective abortion and to identify factors associated with repeat pregnancy in the subsequent year. METHODS: We performed a historical cohort study of 865 women who underwent first-trimester surgical abortion at a hospital-based family planning clinic between March 2003 and February 2004. We collected demographic and clinical data from procedure notes and a computerized hospital patient database. We used univariable and multivariable logistic regression to analyze associations between patient characteristics and repeat pregnancy within 1 year. RESULTS: There were a total of 865 women in our cohort. Of these, 753 (87%) returned to the medical center for care; 485 women returned on time, 268 women returned late, and 112 had no further visit. There were 161 repeat pregnancies; 87 (17.9%) in the on-time group and 74 (27.6%) in the late group (P<.001). There were 85 repeat abortions; 42 (8.7%) in the on-time group and 43 (16.0%) in the late group (P<.001). Multivariable analysis demonstrated that on time follow-up was associated with a reduction in repeat pregnancy (odds ratio [OR] 0.54, 95% confidence interval [CI] 0.37-0.77) and repeat abortion (OR 0.53, 95% CI 0.33-0.86). A total of 130 women received injectable depot medroxyprogesterone acetate (DMPA) at the time of abortion. The rates of repeat pregnancy and abortion in these women were 13.7% and 6.2%, respectively. Women who received DMPA had a decrease in rates of repeat pregnancy (OR 0.41, 95% CI 0.23-0.73) and repeat abortion (OR 0.45, 95% CI 0.20-0.99). CONCLUSION: Timely follow-up and immediate administration of DMPA are associated with a decrease in repeat pregnancy in the 12 months after first-trimester elective abortion. PMID- 19300333 TI - Effect of previous live birth and prior route of delivery on the outcome of early medical abortion. AB - OBJECTIVE: To determine the association between type of previous delivery (vaginal compared with cesarean) on the success of medical abortion with mifepristone-misoprostol in early pregnancy. METHODS: The records of 879 women with intrauterine pregnancies at or before 56 days of gestation who underwent medical abortions were reviewed. Medical treatment consisted of 600 mg mifepristone orally followed 48 hours later with oral misoprostol. An ultrasound examination was performed 14-21 days after treatment, and a successful medical abortion was defined as an empty uterus without surgical intervention. Univariable and multivariable logistic regressions were used to determine risk factors for failure of medical abortion. RESULTS: A total of 797 (90.7%) women had successful medical abortions; 82 (9.3%) had failed medical abortions. Multivariable logistic regression indicated that women with gestational ages greater than 42 days (odds ratio [OR] 2.53, 95% confidence interval [CI] 1.55 4.05) had higher odds of failed abortion compared with a gestational age less than 43 days. Parous women (OR> or =3.94, 95% CI 1.83-8.53) and those with prior cesarean delivery (OR 9.59, 95% CI 4.30-21.39) were more likely to have failed abortions compared with nulliparous women. Among 523 parous women (68 had failed abortion), those with gestational ages greater than 42 days (OR 2.07, 95% CI 1.22 3.50) and prior cesarean delivery (OR 3.33, 95% CI 1.95-5.69) were more likely to have failed abortions compared with those with gestational ages less than 43 days or with prior vaginal delivery. CONCLUSION: Parous women are at increased risk for failed medical abortion in comparison with nulliparous women. Prior cesarean delivery is significantly associated with failed medical abortion. PMID- 19300334 TI - Emerging concepts in antibiotic prophylaxis for cesarean delivery: a systematic review. AB - OBJECTIVE: To review the current status of antibiotic prophylaxis for cesarean delivery, emerging strategies to enhance the effectiveness of antibiotic prophylaxis in reducing postcesarean infection, and the implications of the emerging practices. DATA SOURCES: We conducted a full PubMed (January 1966 to July 2008) search using the key words "cesarean" and "antibiotic prophylaxis." A total of 277 articles were identified and supplemented by a bibliographic search. METHODS OF STUDY SELECTION: We selected a total of 15 studies, which included all published clinical trials, meta-analyses of clinical trials, and observational studies evaluating either the timing of antibiotics or the use of extended spectrum prophylaxis. We also reviewed nine reports involving national recommendations or technical reviews supporting current standards for antibiotic prophylaxis. TABULATION, INTEGRATION, AND RESULTS: We conducted an analytic review and tabulation of selected studies without further meta-analysis. Although current guidelines for antibiotic prophylaxis recommend the administration of narrow-spectrum antibiotics (cefazolin) after clamping of the umbilical cord, the data suggest that antibiotic administration before surgical incision or the use of extended-spectrum regimens (involving azithromycin or metronidazole) after cord clamp may reduce postcesarean maternal infection by up to 50%. However, these two strategies have not been compared with each other. In addition, their effect on neonatal infection or infection with resistant organisms warrants further study. CONCLUSION: The use of either cefazolin alone before surgical incision or an extended-spectrum regimen after cord clamp seems to be associated with a reduction in postcesarean maternal infection. Confirmatory studies focusing additionally on neonatal outcomes and the effect on resistant organisms, as well as studies comparing both strategies, are needed. PMID- 19300335 TI - A proposed model for managing cases of neurologically impaired infants. AB - The current mechanism for obtaining financial support for families with neurologically impaired infants is seriously flawed. It relies on payment awarded through the tort system based on a claim that medical negligence was responsible for the infant's condition. The system is extraordinarily inefficient and expensive, as well as being unfair to many families with affected children and to physicians who are unjustly accused of contributing to outcomes they could not have prevented. Furthermore, the exorbitant malpractice premiums necessary to support the system are threatening the future of obstetric practice in the United States. This article describes a two-pronged program designed to correct these inequities and to assess each case for the occurrence of medical negligence, which has been submitted to the New York State legislature as a proposed bill entitled the Neurologically Impaired Program for New York State (S7748). PMID- 19300337 TI - Thyroid dysfunction in pregnancy: the basic science and clinical evidence surrounding the controversy in management. AB - Maternal hypothyroidism is known to result in neurodevelopmental disorders in offspring, but whether subclinical hypothyroidism results in lower intelligence quotient (IQ) performance in progeny is an area of debate. Animal studies have shown that fetal thyroxine and triiodothyronine are primarily maternally derived before mid gestation. Other animal data reveal that fetal brain damage at a time that is analogous to the first trimester in humans can be linked to irreversible future brain damage. A large study conducted on an unselected population of pregnant women, both with known diagnosis of hypothyroidism and those who were screened but not diagnosed, found a four-point difference in the IQ levels of the offspring, raising the question of clinical significance. The endocrine community has accepted that subclinical hypothyroidism causes a significant decrease in IQ scores and has advocated for routine screening of pregnant women. However, obstetric authorities have cautioned that more research is needed before a causal relationship between subclinical hypothyroidism and lower IQ performance can be verified. Consequently, the American College of Obstetricians and Gynecologists has stated that routine screening and treatment of subclinical hypothyroidism cannot be recommended. We will review the basic science and clinical evidence for the neurodevelopmental effects of thyroid dysfunction in pregnancy. PMID- 19300336 TI - Antenatal testing-a reevaluation: executive summary of a Eunice Kennedy Shriver National Institute of Child Health and Human Development workshop. AB - In August 2007, the Eunice Kennedy Shriver National Institute of Child Health and Human Development, the National Institutes of Health Office of Rare Diseases, the American College of Obstetricians and Gynecologists, and the American Academy of Pediatrics cosponsored a 2-day workshop to reassess the body of evidence supporting antepartum assessment of fetal well-being, identify key gaps in the evidence, and formulate recommendations for further research. Participants included experts in obstetrics and fetal physiology and representatives from relevant stakeholder groups and organizations. This article is a summary of the discussions at the workshop, including synopses of oral presentations on the epidemiology of stillbirth and fetal neurological injury, fetal physiology, techniques for antenatal monitoring, and maternal and fetal indications for monitoring. Finally, a synthesis of recommendations for further research compiled from three breakout workgroups is presented. PMID- 19300338 TI - Can we communicate gravidity and parity better? AB - Communication of gravidity and parity is a typical part of any written or oral presentation related to an obstetric or gynecologic patient. The ostensible purpose of including this information at the beginning of a discussion of a woman's care is to provide a quick context of the patient from a reproductive standpoint. Unfortunately, the systems in use for describing gravidity, parity, and reproductive outcomes are without clear epidemiologic, biologic, or clinical basis. In description of parous events, births and abortions easily can be confused in the gray zone of 20 to 24 weeks, and terminology used often fails to take into account the clinical context. In this article, we discuss the pitfalls of current systems used for describing a woman's reproductive history and recommend new and simple nomenclature to enhance our ability to communicate properly. PMID- 19300339 TI - Confronting the legal risks of prescribing the contraceptive patch with ongoing litigation. AB - Recent changes in U.S. Food and Drug Administration (FDA) labeling and news reports of lawsuits resulting in million-dollar settlements understandably may deter gynecologists from prescribing the transdermal contraceptive patch Ortho Evra (Ortho-McNeil Pharmaceutical, Inc., Titusville, NJ). Gynecologists who, with all good intentions, prescribe an FDA-approved drug such as the contraceptive patch potentially could find themselves liable for an adverse drug reaction. Although much of the current focus by plaintiff attorneys and the news media is on the contraceptive patch, no prescription contraceptive method is without medical risks to the patient or legal risks to the prescribing gynecologist. The purpose of this commentary is to provide an overview of the medical-legal controversies and pitfalls in prescribing the contraceptive patch as well as to outline how gynecologists can avert legal liability by providing proper informed consent. Despite FDA labeling changes and ongoing litigation, with proper informed consent, the contraceptive patch still may be the best choice for many patients who prefer the convenience of a weekly patch over a daily oral contraceptive. Also, regardless of the contraceptive option chosen, the principles of providing and documenting proper informed consent in medical records are applicable not only to providing quality care to patients, but also to protecting the legal interests of the prescribing gynecologist. By documenting proper informed consent in medical records, gynecologists should feel more at ease in prescribing the contraceptive method that best fits their individual patients' needs, even in the presence of ongoing litigation. PMID- 19300340 TI - Maternal-fetal surgery: where are we and how did we get here? AB - Using the criteria proposed in 1982 to justify consideration of in utero surgical treatment of fetal disorders, the development of maternal-fetal treatment is described, and an assessment of current status is provided. Significant issues remain, and many questions are unanswered. By using three different disorders, congenital diaphragmatic hernia, open spina bifida, and twin-to-twin transfusion, the complexity and sophistication of the field are explored, as well as the existing gaps in understanding. PMID- 19300341 TI - Pelvic floor muscle training for prevention and treatment of urinary and faecal incontinence in antenatal and postnatal women. PMID- 19300343 TI - Anti-factor xa plasma levels in pregnant women receiving low molecular weight heparin thromboprophylaxis. PMID- 19300345 TI - Mifepristone for the treatment of uterine leiomyomas: methodological issues and clinical implications. PMID- 19300347 TI - ACOG Practice Bulletin No. 102: management of stillbirth. PMID- 19300348 TI - ACOG Committee Opinion No. 429: health disparities for rural women. PMID- 19300349 TI - ACOG Committee Opinion No. 430: preimplantation genetic screening for aneuploidy. PMID- 19300351 TI - Lemierre's syndrome: a serious complication of an odontogenic infection. AB - Necrobacillosis, postanginal septicaemia or Lemierre's Syndrome is characterised by suppurative thrombophlebitis of the internal jugular vein with embolization to several sites, including the lungs. We report the case of a 38-year-old man who was initially hospitalized because of odontogenic cellulitis. Given the deterioration of his clinical state (septic shock and multiple organ failure), neck computed tomography was performed, revealing both cervical and parotid abscesses, and thrombosis of the right internal jugular vein. Streptococcus salivarius was isolated. The patient was treated with intravenous antibiotics, and surgical drainage and after 6 weeks of treatment, recovered completely. Lemierre's Syndrome is an uncommon, but potentially lethal complication of an odontogenic infection. The case reported here is interesting since the pathogen and the site of primary infection are unusual. Fusobacterium necrophorum is the most common pathogen in Lemierre's Syndrome, and to the best of our knowledge there are no similar case reports with Streptococcus salivarius as the causative bacteria. Early recognition and high-dose antibiotics are critical elements in reducing mortality. PMID- 19300352 TI - Morphometric analysis of the dorsum linguae in patients with Oral Lichen Planus. AB - OBJECTIVE: Morphometric study of the dorsum linguae in patients with Oral Lichen Planus (OLP). MATERIAL AND METHOD: In 236 patients with a clinicopathologic diagnosis of OLP, 111 were found to have involvement of the dorsum linguae. For the purposes of the study, 41 of these cases were used, due to the availability of photographic records fulfilling quality conditions according to objective criteria. The experimental variable was defined as the proportion of affected dorsum linguae measured and calculated using the MIP4(R) computer software. RESULTS: In 236 patients with OLP, the mucosa of the dorsum linguae was affected in 47.6% of the cases (total 111). The proportion of affected area was studied in 41 of these cases, 8 men (19.5%) and 33 women (80.5%). In 73% of these cases, the affected area was less than 50% of the total surface of the dorsum linguae. There were no statistically significant differences (p=0.495) in relation to age and proportion of lingual surface affected. On the other hand, significant statistical differences were found between the period of disease evolution and area of lingual involvement (p=0.044). CONCLUSION: Lingual involvement is frequent in patients with OLP, and manifests as de-papillate areas to the left and right of the median sulcus of tongue (occasionally linked by an isthmus) and having a certain " butterfly wing" symmetry. PMID- 19300353 TI - Type 1 diabetes mellitus and periodontal disease: relationship to different clinical variables. AB - OBJECTIVE: This study is designed to evaluate the frequency of periodontal disease in a group of patients with type 1 diabetes mellitus and how this relates with diabetes metabolic control, duration of diabetes, and presence of diabetic complications. METHODS: A comparison was made of periodontal parameters (plaque index, bleeding index, pocket depth and attachment loss) in a group of diabetic patients (n=90) versus a group of non-diabetics (n=90). Logistic regression analysis was performed to evaluate relationship between periodontal parameters and degree of metabolic control, the duration of the disease, and the appearance of complications. RESULTS: Diabetics had greater bleeding index (p< 0.01), deeper periodontal pockets (p< 0.01) and more periodontal attachment loss (p< 0.01) than non-diabetics. Deficient metabolic control and presence of diabetic complication were associated with higher bleeding index and pocket depth (p 0 ). Dental erosion results were linked to data supplied by a questionnaire assessing socio-demographic characteristics, behaviour related to alcohol and drug use, including a history of drug and alcohol abuse, and oral health promotion using logistic multivariate regression analysis. RESULTS: Enamel and/or dentine erosion lesions were present on 49.4% of the teeth. Among these, 36.9% of occlusal surfaces presented dental erosion with a severity level of 1, 11.4% with a severity level 2 and 1.1% with a severity level 3. The highest occurrence of severity level 3 was found to exist in palatal dental surfaces (1.9%). Dental erosion on teeth surfaces was found to be independently associated with intra oral location (by arch, tooth type), patients' socio-demographic characteristics, behaviour associated with a history of alcohol and drug abuse, and oral health promotion. Maxillary teeth, more so than mandibular teeth, presented moderate to higher severity erosion injuries (Wilcoxon test; p< 0.001); and significant differences in the severity of dental erosion were found between anterior and posterior teeth, in both the maxillary and the mandibular arches (Friedman test, p< 0.001). CONCLUSIONS: Alcohol-dependent patients undergoing a detoxification programme presented a high occurrence and a low severity of dental erosion lesions.The palatal surfaces of the anterior teeth, followed by incisive/occlusal surfaces in both anterior and posterior teeth, respectively, were most affected by erosion injuries. PMID- 19300356 TI - Oral lesions in 166 patients with cutaneous psoriasis: a controlled study. AB - OBJECTIVES: This study was aimed to test if the frequency of oral lesions bears statistical correlation or not with the condition of cutaneous psoriasis. STUDY DESIGN: Two groups were examined, one made up of 166 patients with skin psoriasis and the other with the same number of individuals with a negative history of skin diseases (control group), matched by age, race, and sex. Patients with psoriasis were grouped according to their having localized or generalized forms of the disease. The oral mucosa was thoroughly examined in both groups. Data were analyzed using chi-square test, Fisher's test, the odds ratio (OR) with a 95% confidence interval (CI), and the Ryan-Holm step-down Bonferroni procedure. The overall significance was set at P < or = 0.05. RESULTS: The oral lesions significantly associated with psoriasis were fissured tongue (FT, OR = 2.7; 95% CI: 1.3-5.6), and geographic tongue (GT, OR = 5.0; 95% CI: 1.5-16.8). Other factors analyzed, such as topical and/or systemic medication for treatment of psoriasis versus nontreated patients, and localized versus generalized forms of psoriasis presented no statistical association with the frequency of FT or GT lesions (p>0.05). CONCLUSIONS: Patients with psoriasis presented no specific oral lesion different from those seen in the control group. Although further investigation is warranted to establish whether or not either FT or GT can be characterized as an oral expression of psoriasis, the present investigation did find for both these types of lesions that the frequency of each bore a statistically significant relation with the presence of cutaneous psoriasis. PMID- 19300357 TI - Transalveolar screw: a new concept for orthodontic anchorage. AB - The purpose of this article is to describe the use of a new trans-alveolar screw (TAS) as a temporary orthodontic anchorage device for the posterior maxilla, to intrude overerupted maxillary molars. To date, five consecutive patients have been treated with these newly designed screws. Intrusions achieved ranged from 2.1 and 6mm (mean 4.7 mm). The TAS is cheap, easy to place and remove by the orthodontist, has bicortical anchorage, and is loaded on both sides. The main advantage of TAS is that when placed in the maxilla to intrude upper molars, it allows application of intrusive force from both buccal and palatal aspects simultaneously, so that the line of force in relation to the center of resistance of the posterior segment, permits an in-mass intrusion, avoiding buccal tipping or rotations. Moreover the surgical procedure for inserting and removing the bicortical screw is simple and does not require any surgical flap, so complications are minimal and screws can be loaded immediately, without requiring any waiting healing period of time. PMID- 19300358 TI - Presence of fibronectin peptides in saliva of patients with Sjogren's syndrome: a potential indicator of salivary gland destruction. AB - OBJECTIVE: The purpose of this preliminary study was to monitor the degree of destruction of salivary glands in Sjogren disease by the detection of fibronectin peptides in patients' saliva. STUDY DESIGN: The sample consisted of 10 subjects divided in 2 groups, one with Sjogren disease and a control group. Saliva samples were submitted to an inmunodetection analysis. In addition, non pathological salivary glands, obtained from 2 subjects who underwent minor oral surgery, were incubated with leukocyte homogenates and analysed to compare the obtained fragments. RESULTS: The inmunodetection analysis of Sjogren saliva revealed multiple protein bands, including fibronectin, that were not present in saliva from healthy individuals. The inmunostained bands varied depending on the course of the disease, showing more fibronectin fragments in an active phase. Furthermore, results obtained from the non-pathological glands incubated with leukocyte homogenates were similar to those revealed in saliva from Sjogren patients. CONCLUSION: The presence of fibronectin peptides in Sjogren patients' saliva can constitute a method to monitor activity in Sjogren's disease. PMID- 19300359 TI - Surgical ciliated cyst of the maxilla. Clinical case. AB - Surgical ciliated cyst is uncommon in Western countries but frequently reported in Asian populations as a delayed complication of surgery, with inclusion in the bone of nasal or sinus mucosa. Isolated cases have also been reported in the mandible after orthognathic surgery. We report a case in the maxillary region three years after radical sinus surgery for chronic sinusitis. Intraoral examination revealed a small painless tumefaction with no color change in the surrounding tissues. Computed tomography demonstrated a well-defined intraosseous lesion lateral to the maxillary sinus. The lesion was completely excised, when histology demonstrated a pseudostratified ciliated epithelial lining. Differential diagnosis from other lesions such as an odontogenic keratocyst or inflammatory cyst were also made. This case report,uncommon in Western countries, of a surgical ciliated cyst illustrates the need for a meticulous surgical technique, proper management of complications, and routine, long-term follow-up of patients undergoing any type of sinus surgery. PMID- 19300360 TI - Multiple supernumerary teeth not associated with complex syndromes: a retrospective study. AB - OBJECTIVES: To determine the epidemiology and describe the clinical and radiographic characteristics, the type of treatment, and the possible delayed appearance of new supernumerary teeth in patients with non-syndromic multiple hyperdontia. PATIENTS AND METHODS: We conducted a small retrospective observational study of 8 patients diagnosed with non-syndromic multiple hyperodontia. Multiple hyperdontia not associated to complex syndromes was defined as apparently generally healthy patients with one or more supernumerary teeth in two or more areas. RESULTS: The average patient age was 16.23 years; males predominated (3:1). Multiple hyperodontia with a minimum of 2 and a maximum of 9 supernumerary teeth was found (total: 34 mean: 4.25). The most frequent location was the upper jaw (76.47%). Eumorphic teeth were seen at lower premolar level, while the rest were all heteromorphic. There was altered eruption of the contiguous teeth of 4 of the impacted supernumerary teeth all the rest being asymptomatic. Extraction was the treatment in all patients, and in one of them the delayed appearance of 4 supernumerary teeth was detected. CONCLUSIONS: Multiple hyperodontia rarely occurs without being associated with complex syndromes. Prophylactic surgical removal of the supernumerary teeth is generally the treatment of choice. PMID- 19300361 TI - Ellis-van Creveld syndrome. Case report and literature review. AB - Ellis-van Creveld syndrome is a genetic disorder that was first described by Richard Ellis and Simon van Creveld in 1940. The four principal characteristics are chondrodysplasia, polydactyly, ectodermal dysplasia and congenital heart defects. The orofacial manifestations include multiple gingivolabial musculofibrous fraenula, dental anomalies, hypodontia and malocclusion. The disease can be diagnosed at any age, even during pregnancy. The differentiation should be made between Jeune syndrome and other orofaciodigital syndromes. PMID- 19300362 TI - Oral lichen planus and oral lichenoid lesions; a critical appraisal with emphasis on the diagnostic aspects. AB - Oral lichen planus (OLP) has a prevalence of approximately 1%. The etiopathogenesis is poorly understood. The annual malignant transformation is less than 0.5%. There are no effective means to either predict or to prevent such event. Oral lesions may occur that to some extent look like lichen planus but lacking the characteristic features of OLP, or that are indistinguishable from OLP clinically but having a distinct cause, e.g. amalgam restoration associated. Such lesions are referred to as oral lichenoid lesions (OLLs). The management of OLP and the various OLLs may be different. Therefore, accurate diagnosis should be aimed at. PMID- 19300363 TI - Orofacial features of Treacher Collins syndrome. AB - Treacher Collins syndrome (TCS) is a rare autosomal dominant disorder of craniofacial development. Major features include midface hypoplasia, micrognathia, microtia, conductive hearing loss, and cleft palate. The present study is on the orofacial features of 7 Brazilian patients with sporadic TCS aged 4 to 38 years. All patients presented the typical down-slanting palpebral fissures, colobomas, zygomatic and mandibular hypoplasia, partial absence of the lower eyelid cilia, and abnormalities of the ears. Malocclusion was present in all patients, and an anterior open bite was found in 3 patients. None of the patients had a cleft palate. PMID- 19300364 TI - Sjogren's syndrome of the oral cavity. Review and update. AB - Sjogren's syndrome is one of the most frequent autoimmune diseases. It is a chronic and systemic disorder predominantly found in women, and is characterized by the appearance of a lymphocytic inflammatory infiltrate, with dryness of the oral cavity and eyes, secondary to involvement of the salivary and lacrimal glands. The underlying causal mechanism involves a number of factors and has not been clearly established, though an autoimmune response is known to be triggered, with the accumulation of immune complexes in the gland acini that interfere with gland function. In the oral cavity, xerostomia or hyposialia is the most disabling manifestation for patients, and is accompanied by rapidly progressing caries, candidiasis and an important worsening of buccodental health. The most important complication is a 44-fold increase in the risk of developing non Hodgkin lymphoma, compared with the general population. The treatment of Sjogren's syndrome is limited to symptomatic management, and involves the use of solutions to replace salivary secretion and afford a measure of hydration, cholinergic agents such as pilocarpine to stimulate the unaffected gland tissue and, recently, the administration of substances that act against surface antigens of the B lymphocytes, such as anti-CD20 and anti-CD22 antibodies. The present study provides an update on this disease, placing special emphasis on its odontologic implications. PMID- 19300365 TI - Oral Lichen Planus: a retrospective study of 420 Iranian patients. AB - OBJECTIVE: In terms of the demographic and clinical characteristics, this is the one of the largest studies on Iranian patients with Oral Lichen Planus (OLP). STUDY DESIGN: Data was taken from the medical records of 420 consecutive patients referred to the Oral Medicine Department, and who were subsequently found to have clinical and usual histopathology consistent with features of OLP. RESULTS: Seventy percent of the patients had been referred to the Oral Medicine Department by general dental practitioners. 52.6% were referred due to oral mucosal and/or gingival pain or burning sensation. Reticular OLP was the most common presentation (76.9%); about 18% of patients reported symptoms or signs, or had a known history of OLP, or possible Lichen Planus affecting non-oral epithelia. A malignant transformation rate of 0.07% was observed. PMID- 19300366 TI - Relationship between mandibular fracture and impacted lower third molar. AB - Many studies have shown that the greatest risk of fracture of the mandibular angle is related to the presence of an unerupted lower third molar, based on the hypothesis that there is a decrease in the area of bone and absorption of the impact in this area, leading some surgeons to indicate its prophylactic removal in patients most exposed to the risk factors of facial trauma. On the other hand, other authors have observed a greater frequency of condylar fractures in patients without an impacted lower third molar. PURPOSE: The aim of this study was to relate the condylar and angle fracture with an unerupted lower third molar, taking into account the position of the tooth. MATERIAL AND METHODS: Panoramic radiographs were used to determine if the presence or absence of the third molar is related to the occurrence of mandibular fractures, such as angle and condylar fractures. RESULTS: In a total of 43 patients with angle fractures, the greatest percentage had erupted teeth, and 41.9% had impacted teeth; however there was no significant difference between the fractured side and the tooth condition (p=0.350). There were 91 condylar fractures and in 42.9% the third molar was absent and in 40.7% the tooth was erupted . There was no significant difference between the fractured side and the tooth condition (p=0.852). CONCLUSIONS: The absence of an impacted third molar may increase the risk of condylar fractures and decrease the prevalence of mandibular angle fractures. PMID- 19300367 TI - The post-mortem pink teeth phenomenon: a case report. AB - This study presents the case of the post-mortem pink teeth phenomenon observed during an autopsy procedure performed on the body of a man who was kidnapped and murdered approximately 30 days before the examination. The corpse was in an advanced stage of decomposition and putrefaction. Both maxillary and jaw bones were intact, as well as the permanent teeth which presented the "pink teeth phenomenon", probably due to a haemorrhage in the pulp chambers. The pink discolouration was most pronounced at the neck of the teeth. The cause of death was asphyxia. Although the examiners stressed that post-mortem pink teeth must not be considered as a reliable odontological parameter for determining the cause of death, the results of other studies have shown that the pink teeth phenomenon is a common finding related to cases of asphyxia such as strangulation, drowning or suffocation. Thus, the pink teeth phenomenon must be studied in order to determine its role as a post-mortem finding. As of now, an exact relationship between the cause of death and this phenomenon remains unknown. PMID- 19300368 TI - Expression of pro-inflammatory protein, iNOS, VEGF and COX-2 in oral squamous cell carcinoma (OSCC), relationship with angiogenesis and their clinico pathological correlation. AB - One main etiology for oral squamous cell carcinoma (OSCC) is inflammation. Inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) are the important molecules showing close relation to not only inflammation but also carcinogenesis and angiogenesis. Angiogenesis is defined as the formation of new blood vessels from existing vasculature. It is necessary for tumor growth and progression and also involved in metastasis. The objective of this research was to study the expression and relationship among iNOS, VEGF, COX-2, angiogenesis and their clinico-pathological correlation in OSCC. In this study, standard indirect immunohistochemical technique using polyclonal antibodies specific to human iNOS, VEGF, COX-2 and CD31 was performed in formalin-fixed paraffin-embedded tissue sections of 66 OSCC samples. The staining patterns and intensity are measured and analyzed statistically. The results showed that epithelial components of squamous cell carcinomas demonstrated moderate to intense staining for iNOS, VEGF and COX-2. iNOS shows correlation with cervical lymph node metastasis and tumor staging (TNM) of the patients and angiogenesis. VEGF shows correlation with tumor grading, tumor staging and angiogenesis. COX-2 shows correlation with cervical lymph node metastasis. In conclusion, the expression of iNOS, VEGF and COX-2 exists in OSCC. The data provided show the expression of these chemical mediators associated with carcinogenesis and angiogenesis in OSCC. It can be the primary database before using angiogenesis drug against these mediators for OSCC treatment. PMID- 19300369 TI - A comparative study of the long term results of root coverage with connective tissue graft or enamel matrix protein: 24-month results. AB - BACKGROUND: The objective of this study was to compare the 24 month results of coronally advanced flap + enamel matrix derivates (EMD+CAF) and CAF+ connective tissue graft (CTG+CAF) in the treatment of Miller Class I recession defects. METHODS: Twelve patients with bilateral gingival recessions were treated with EMD+CAF or CTG+CAF. Vertical recession depth (VRD), keratinized tissue width (KTW), clinical attachment level (CAL), and clinical probing depth (CPD) were measured preoperatively, 1 and 2 years post surgery. A paired t-test and independent t-test were used to compare differences for the measured characters within and between groups, respectively. RESULTS: After 24 months, a significant decrease in VRD was observed in CAF + EMD (3.33+/- 0.30 mm) and CAF + CTG (4.5 +/ 0.28 mm) treated sites. There was also a significant increase in KTW (0.83+/- 0.23 mm versus 2.08+/- 0.14 mm in EMD+CAF and CTG+CAF sites, respectively). The gain in CAL was 3.54 +/- 0.38 mm and 4.45+/- 0.30 mm in the EMD+CAF and CTG+CAF groups, respectively. There were significant differences between the treatments for VRD, CAL, and KTW at the end of study. CONCLUSIONS: The CTG+CAF procedure seems to provide better long-term results than the EMD+CAF in obtaining root coverage, increasing the KTW and CAL gain. PMID- 19300370 TI - Erupted odontomas: a report of three cases and review of the literature. AB - INTRODUCTION: Odontomas are benign odontogenic tumors composed of enamel, dentine, cement and pulp tissue. They are usually clinically asymptomatic, but often associated with tooth eruption disturbances. In exceptional cases the odontoma erupts into the mouth . The present study reports three cases of odontomas erupted into the oral cavity and reviews the literature. CLINICAL CASES: The first case was an 11-year-old girl with impacted 4.6 associated to a radiopaque mass measuring about 2 cm in diameter. The histological diagnosis was complex odontoma. A clinical and radiological follow-up of 4.6 was carried out until its spontaneous eruption. The second case was a 26-year-old male presenting a hard, yellowish-brown mass located distal to the upper left second molar. Computed tomography confirmed the presence of a radiopaque lesion, and the histopathological study confirmed a complex odontoma. The third patient was a 27 year-old male reporting tongue irritation due to tooth eruption in the inferior lingual region. A periapical radiograph revealed a mixed radiopaque lesion associated to impacted 3.2. The histological report in this case indicated a compound odontoma. DISCUSSION: Odontomas erupting into the oral cavity are rare. The first case was published in 1980, and since then only 17 cases have been reported in the literature. Eight of the 17 cases were complex odontomas; the rest were compound odontomas. Pain, swelling and infection were the most common symptoms, and 13 cases presented an impacted tooth associated with the lesion. PMID- 19300371 TI - Oral implant rehabilitation in a patient with Moebius syndrome. AB - INTRODUCTION: Moebius syndrome is a rare congenital disorder characterized by unilateral or bilateral involvement of the sixth and seventh cranial nerves, resulting in a lack of facial expression and eye movements. These patients suffer a series of oral manifestations that may complicate their dental treatment, such as facial and tongue muscle weakness, uncontrolled salivation secondary to deficient lip sealing, micrognathia, microstomia, bifid uvula, gothic and fissured palate, fissured tongue, and glossoptosis. The underlying etiology remains unclear, though vascular problems during embryogenesis appear to be involved. CLINICAL CASE: We report the case of a woman with Moebius syndrome and total edentulism . Eight years ago she underwent complete oral rehabilitation with the placement of two implants in each dental arch. DISCUSSION: Moebius syndrome has still an unknown etiology, although it is related to disorders during pregnancy. This kind of patient can be rehabilitated using oral implants. PMID- 19300372 TI - Oral bisphosphonates-associated osteonecrosis in rheumatoid arthritis. AB - Adverse effects associated with the use of bisphosphonates are infrequent and consist of pyrexia, renal function impairment, and hypocalcemia. Bisphosphonates associated osteonecrosis of the jaws is an uncommon but potentially serious complication of intravenous bisphosphonate therapy in cancer patients. The degree of risk for osteonecrosis in patients taking oral bisphosphonates, such as alendronate, is uncertain and warrants careful monitoring. Oral bisphosphonates associated osteonecrosis can occur in patients with rheumatoid arthritis. We report a case of mandibular osteonecrosis in a patient who received alendronate for 3.8 years. The pathology improved after bisphosphonate therapy discontinuation and sequestrectomy. To our knowledge there are only three cases published in the literature relating bisphosphonates-associated osteonecrosis of the jaws in patients with rheumatoid arthritis. All the cases published, including our case, have reported association between methotrexate, prednisone and alendronate sodium (Fosamax) therapy. Corticosteroid therapy and dental surgery could increase the risk of developing bisphosphonates-associated osteonecrosis of the jaws in these patients. PMID- 19300373 TI - Proliferative Verrucous Leukoplakia--diagnostic pitfalls and suggestions. PMID- 19300374 TI - Central mucoepidermoid carcinoma: report of a case with 11 years' evolution and peculiar macroscopical and clinical characteristics. AB - Central mucoepidermoid carcinomas (CMC) are uncommon tumours, comprising 2-3% of all mucoepidermoid carcinomas reported. They have been reported in patients of all ages, ranging from 1 to 78-years, with the overwhelming majority occurring in the 4th and 5th decades of life. They are histologically low-grade cancers, usually affecting the mandible as uniocular or multiocular radiographic lesions. The authors report a case of CMC of the mandible with a long evolution, and peculiar clinical and macroscopical features related with the long term evolution of the disease. A 53-year-old male patient had expansion of buccal and lingual cortices of the anterior region of the mandible, covered by ulcerated mucosa, with 11 years evolution. An incisional biopsy was performed, and the histopathological findings confirm low-grade mucoepidermoid carcinoma. The patient was treated with a mandibulectomy, followed by supraomohyoid neck dissection. There was no evidence of local recurrence, regional or distant metastasis revealed; and the patient was alive and without disease after a follow up interval of 36 months. PMID- 19300375 TI - Artifacts in brain magnetic resonance imaging due to metallic dental objects. AB - OBJECTIVE: Artifacts caused by metallic objects, such as dental crowns, dental implants and metallic orthodontic appliances, are a common problem in head and neck MRI. The aim of this retrospective study was to identify the main metal dental objects that produce artifacts on brain MRIs. STUDY DESIGN: Imaged metallic artifacts and their sources were identified. Artifact image plane was rated on a score of 0 or 1 (0--distinguishable for diagnosis and 1--not distinguishable for diagnosis). RESULTS: Seventy-eight percent of the artifacts appearing in images were caused by orthodontic appliances, followed by dental titanium implants (18%) and metallic crowns (4%). Orthodontic appliances obtained the highest scores in all planes. CONCLUSIONS: We concluded that is difficult to avoid the effect of metallic artifacts in the maxillofacial regions on brain scan. Removing metallic parts of the orthodontic appliance should ensure diagnostically useful quality scans. PMID- 19300376 TI - Prevalence and distribution of oral mucosal lesions in an adult Turkish population. AB - OBJECTIVES: The purpose of this study was to assess the prevalence and distribution of oral mucosal lesions in a Turkish adult population. MATERIAL AND METHODS: Five thousand consecutive patients were examined during routine dental treatment. Some of the mucosal changes were diagnosed solely by clinical examination. When clinical features were not diagnostic, a biopsy was undertaken. Chi-square test was used to analyze the data. RESULTS: The overall incidence of oral mucosal changes or lesions was 15.5%. The lesions were classified as anatomic changes, ulcerated lesions, tongue lesions, white lesions, benign lesions, color alterations, and malignant lesions. Anatomic changes (7%), ulcerated lesions (6.6%), and tongue lesions (4.6%) were the most common lesions. White lesions were observed in 2.2% of all patients. Among the white lesions, leukoplakia was identified in men 4 times more frequently than it was in women. Benign lesions and color alterations were identified in 1.6% and 1.2% of all patients, respectively. In addition, 3 patients (0.06%) were diagnosed as having squamous cell carcinoma, and 1 patient (0.02%) was diagnosed as having adenocarcinoma. There was a statistically significant relation between smoking and the occurrence of mucosal lesions whereas no relation was found between alcohol consumption and mucosal lesion occurrence. Besides, no relation was found between systemic diseases and oral mucosal lesion occurrence. CONCLUSIONS: This study has provided information about the epidemiologic aspects of oral mucosal lesions that may prove valuable in planning of future oral health studies. PMID- 19300377 TI - Prevalence of medically compromised conditions in dental patients. AB - OBJECTIVES: To study the prevalence of medically compromised conditions in patients seeking dental treatment at the Faculty of Dentistry, Chulalongkorn University. STUDY DESIGN: The files of 58,317 patients seeking dental treatments at the Faculty of Dentistry, Chulalongkorn University from 2002-2004 were retrieved and reviewed for medically compromised conditions. RESULTS: From a total of 58,317 patients, 7,167 patients (12.2%) had medically compromised conditions. The five most prevalent conditions in descending order were allergy, hypertension, diabetes mellitus, heart diseases and thyroid diseases. CONCLUSIONS: Even though the prevalence of medically compromised conditions in dental patients is not high, dentists should bear in mind that some of the patients may harbor such conditions which are contraindicated for certain dental procedures or medication or require special attention when treating these patients. Therefore, a thorough history taking and careful clinical examinations are mandatory before commencing any dental treatment. PMID- 19300378 TI - Primary melanoma of the oral cavity: ten cases and review of 177 cases from literature. AB - Oral melanoma is an infrequent but aggressive neoplasm. In contrast to cutaneous melanoma, it does not have a defined set of clinical and pathologic classification criteria. We present 10 cases of primary melanoma of the oral mucosa, and review 177 cases published in the international literature. PATIENTS AND METHODS: We compiled a database of patients seen at our department between years 1990 and 2004. A retrospective study of this data was performed. Age, sex, localization and histopathology were all considered. Tobacco consumption and the presence of traumatic factors in the oral cavity were also compared. We compiled another database with 177 cases reported in the international literature during the same time period. Primary and metastatic cases were included. Age, sex and localization were among the factors taken into account. RESULTS: Of the total of 23,685 patients seen at our department in this period, 399 (1.68%) were diagnosed with cancer, 10 (2.5%) of whom were diagnosed with primary oral melanoma. The gender distribution was 1:1, and ages ranged from 30 to 88 years old, with a mean of 67.5 years old. In 80% of the cases, the oral melanoma was localized in the maxillary, and in 20% of the cases, it was localized in the mandible. No connection was found with tobacco consumption. In 60% of the cases, we believe there was a link with prosthetic microtrauma. Of the 177 cases described in the literature, 89% were primary and 11% secondary. The gender distribution was 46.9% female and 53.1% male. Ages ranged from 16 to 91 years old, with a mean of 59.2 years old. Localization: 68.36% maxillary, 11.30% mandible and 20.34% in various oral localizations. PMID- 19300379 TI - Traditional Chinese Medicine in treatments to depression. AB - Traditional Chinese Medicine (TCM) has a long history of more than 5,000 years and it has the earliest medicine book in the world: Yellow Emperor's Canon: Internal Medicine (Huang Ti Nei Jing), by the Yellow Emperor of China (2695-2589 BC) (Lin , 1988; Shampo and Kyle, 1989; Wang, 1999). However the combinational and systemic recipes that bring TCM high efficiency also block the promotion of TCM treatment to various diseases, including depression. In this short comment we firstly give some theoretical basis for TCM treatment to depression, then some clinical reports on efficiency; we also present some possible explanations on TCM treatments to depression. PMID- 19300380 TI - The epidural steroids in the prevention of epidural fibrosis: MRI and clinical findings. AB - BACKGROUND: Epidural fibrosis (EF) represents a frequent and poorly manageable complication of lumbar disk surgery. OBJECTIVES: To investigate the influence of perioperative Epidural Steroid (ES) application on the development of EF. METHODS: One hundred and seventy eight patients underwent L4/5 or L5/S1 discectomy. The study group receiving ES comprised of eighty five patients, and a further control group comprising eighty two patients received a placebo. At a 12 month follow-up, all subjects underwent contrast magnet resonance imaging of the lumbosacral spine. One hundred and sixty seven patients filled in a predetermined questionnaire containing the Visual Analogue Scale (VAS; pain scale) during the first postoperative days and 12 months after. Intergroup differences were analysed and a correlation between the extent of EF and VAS was examined. RESULTS: The groups did not differ regarding the extent of EF. There was a statistically significant correlation between the degree of fibrosis and VAS (P<0.05). However, there was no significant difference in subjective pain assessment between both groups 12 months postoperatively. The application of ES did not influence their return to work. Patients receiving ES experienced less pain on the first and third days after surgery. The average hospital stay after surgery was shorter in the steroid treated group (4.5 days) compared to 5.2 days in the control group (p<0.05). CONCLUSIONS: The application of ES did not prove to be useful in the prevention of Failed Back Surgery Syndrome and epidural scar formation. Postoperative pain was decreased in the steroid treated group during the first postoperative week, but not 12 months postoperatively. PMID- 19300381 TI - Suicidal behavior in children and adolescents: does a history of trauma predict less severe suicidal attempts? AB - OBJECTIVES: The aim of this study was to identify risk factors and possible predictors of severity of suicidal behavior of children and adolescents. METHODS: Seventy-seven patients (15 boys and 62 girls) aged 15.5+/-1.6 years on average, hospitalized due to a suicidal attempt in the department of pediatric psychiatry, were examined. Structured interviews with patients and their parents were used to clinically assess circumstances of suicidal behavior, relevant risk factors and severity of suicidal behavior. RESULTS: The results indicated that patients with any previous traumatic experience tended to have somatically less severe suicidal attempts (p=0.050). Intensity of suicidal intent was associated with a history of depression (p=0.014) and anxiety disorders (p=0.004), and the current stress from a mental disorder (p=0.014). Somatic severity of suicidal behavior was significantly associated with intensity of suicidal intent (p=0.014). A history of any trauma (previous traumatic experience predicted less severe suicidal behavior, p=0.053) and the current stress from sexual problems (p=0.067) were identified as predictors of somatic severity of suicidality. These two predictors showed only a trend level of significance. The only significant predictor of intensity of suicidal intent was the current stress from a mental illness (p=0.017). CONCLUSIONS: Several risk factors of somatic severity of suicidal behavior and intensity of suicidal intent were described. The most important finding of the study was the association between a history of psychological trauma and a tendency to have less somatically severe suicidal behavior. PMID- 19300382 TI - Soluble HLA-G changes in maternal blood serum during the progression of labor. AB - BACKGROUND: The expression of the HLA-G antigen from implantation toward term is crucial for the growth of a semiallogenic fetus as it shields this fetus from the maternal cytotoxic response. Little is known, however, about the potential role of soluble HLA-G isoforms during delivery. The initiation of labor is associated with a complex molecular response leading to a brief activation of the maternal immune system with an accompanying capacity to restrict this activation, and HLA G seems to be an important factor in enabling the proper immune response at the maternal fetal interface. METHODS: In our study the levels of soluble HLA-G concentration were evaluated in the blood serum samples obtained from 47 pregnant women who either underwent cesarean sections or delivered vaginally. The patients were divided into three subgroups according to the progression of labor at the time of the cesarean or, in cases of vaginal delivery, according to the duration of the pregnancy. RESULTS: We have observed that the progression of labor is associated with a continuous increase in the sHLA-G plasma level. The sHLA-G levels were statistically significantly higher in the blood sera obtained from the women in advanced labor than from the women who were at the beginning of labor. CONCLUSION: The changes in sHLA-G concentration levels observed during the stages of labor may indicate that this isoform participates in maintaining reproductive tract homeostasis. PMID- 19300383 TI - Sleep apnea in carotid stenosis. AB - OBJECTIVES: Sleep apnea is associated with advanced atherosclerosis. This study was focused on sleep breathing in patients with hemodynamically significant carotid stenosis, currently free from clinical symptoms. DESIGN AND SETTINGS: 17 patients with carotid artery stenosis of 70% and more in the absence of actual neurological symptoms indicated for non-acute endarterectomy, and 17 age- and sex matched controls were examined using sleep polygraphy. 12 patients had a follow up sleep polygraphy a month after the surgery. RESULTS: The criteria of obstructive sleep apnea (OSA) were met by 4 patients prior to operation, by 2 patients after the operation, and by 2 control subjects. The pre-surgery apnea/hypopnea index (AHI) was 14 (+/-SD=17.0), post-surgery 8.3 (+/-9.0) and in the controls 6.7(+/-6.7). The pre-surgery oxygen desaturation index was 20.1 (+/ 17.7), post-surgery 15.0 (+/-12.0) and in the controls 11.6 (+/-6.1). A comparison between the pre-surgery results seen in the patients and in controls after adjustment for BMI revealed no significant difference. The only significant difference between the pre-surgery and post-surgery values was found in the AHI (P=0.045). CONCLUSION: According to this study there exists an association between carotid stenosis and OSA, however this association is explainable by a higher BMI. The study also found a tendency toward OSA alleviation in response to endarterectomy. PMID- 19300384 TI - Repeated occurrence of clozapine-induced myocarditis in a patient with schizoaffective disorder and comorbid Parkinson's disease. AB - Myocarditis is a rare but life threatening adverse effect of clozapine. Some symptoms of myocarditis--elevated temperature, tachycardia and fatigue--appear commonly during the onset of treatment with clozapine and during the dose titration. We present a case of a patient with concurrent schizoaffective disorder and Parkinson's disease, who twice developed clozapine-induced myocarditis. All symptoms disappeared after the discontinuation of the drug. Early diagnosis, discontinuation of clozapine and supportive therapy of myocarditis lower the risk of a fatal outcome. PMID- 19300385 TI - Pregnancy after organ transplantation. AB - Ovarian function with regular menstrual cycles is usually restored in women of reproductive age after solid organ transplantation. The number of pregnancies reported in these patients increases gradually. Pregnancy is always considered high risk, and not properly planned may lead to serious complications. The best for the patient is to conceive in a period of good general health and good stable graft function, after appropriate preparation and not later than five years after transplantation. Immunosuppressive regimen should be modified before conception. Sirolimus and mycophenolate mofetil should be excluded. The blood levels of immunosuppressive agents should be regularly controlled during the whole pregnancy. The rate of successful pregnancies isn approximately 95% in graft recipients. Increased incidence of preterm labor, anemia and intrauterine growth restriction is observed compared with general population. Organ transplantation itself is not an indication for cesarean section and vaginal delivery is recommended as the best for the patient, the graft and the newborn. Breast feeding is believed to be contraindicated in women on immunosuppressive therapy, however no adverse effects were reported in children of graft recipients who decided to breast feed. The rate of congenital malformations in newborns is approximately 3-4% and does not differ from the rate seen in general population. The rate of perinatal deaths decreased beneath 0.8% in recent reports. Jaundice, hyperglycemia and hyperkalemia, observed more frequently in newborns of graft recipients, are mild and in most cases do not have any clinical implications. PMID- 19300386 TI - Positive effect of long-term tailored physical activity therapy on diabetes mellitus management in an obese type 2 diabetic patient. AB - The case history of a 44-year-old, severely obese Caucasian man with serious decompensation of diabetes and subsequent withdrawal of high-dose insulin is reported. Insulin was withdrawn after 5 years of continuous therapy and the patient was treated for 6 months with diet, individualized physical therapy and oral antidiabetic drugs from the group of insulin sensitizers. Six months later, oral antidiabetics were discontinued for low blood glucose levels, and the treatment was limited to therapeutic life style changes. After insulin withdrawal, the patient's condition markedly improved: he showed better diabetes control, decrease in insulin resistance, weight loss, reduced BMI, reduced waist circumference, sagittal abdominal diameter and body fat, increased fitness level, normalized blood pressure and decreased heart rate at rest. PMID- 19300387 TI - Novel and simple behavioral paradigm for assessing anxiety in rats: effect of diazepam. AB - OBJECTIVES: Anxiety is an emotional state experienced by people, and is not readily modeled in animals. In order to extend till now ethologically derived paradigms used in the evaluation of anxiety and fear in rodents, a modified open field was designed. METHODS: Spontaneous behavior of male rats was investigated in the elevated arena; the bottom was divided by inter-space of different width (2, 4, 6, 8 and 10 cm) into two identical parts. Anxiolytic effects of diazepam (DZP) at doses 0.5, 1.0, 2.5 and 5.0 mg/kg were investigated in the newly designed device and compared with the effects of similar doses in a large circular open-field arena. RESULTS: In Experiment 1 the progressive extension of the inter-space prolonged the first passing, decreased the total number of passing, and increased the inter-space sniffing in intact animals. In Experiment 2 DZP at doses 1.0, 2.5 and 5.0 mg/kg significantly enhanced the readiness to cross, the frequency of passing the inter-space and decreased inter-space sniffing as compared to controls. In Experiment 3 we found that DZP at doses 0.1 and 0.3 mg/kg increased behavioral activity both along the perimeter and in the center of the arena, thus indicating lower level of anxiety. CONCLUSION: The presented modified open-field test is a useful paradigm to investigate risk assessment behavior in rats, and may provide a sensitive novel model of anxiety and fear level. PMID- 19300388 TI - Missing evidence for the effect one-week phytoestrogen-rich diet on mental rotation in two dimensions. AB - INTRODUCTION: It is well known that men outperform women in tests of spatial cognition, such as mental rotation, and that performance in these tasks is influenced by sexual hormones. Phytoestrogens are plant substances chemically similar to estradiol, capable of binding estrogen receptors. In a previous study, one-week phytoestrogen-rich diet improved performance in mental rotation in women, but it remained unclear whether this was due to direct binding of phytoestrogens to estrogen receptors or by modulating testosterone blood levels. OBJECTIVES: We investigated whether one week consumption of phytoestrogens will equally affect performance in mental rotation in women and men. We expected improved performance in women, but unchanged skill in men, if the effects were mediated by changes of testosterone level. On the other hand, direct modulation of estrogen receptors should yield comparable improvement in both genders. METHODS: Thirty-six healthy adult young volunteers (16 females) were divided into the control and soy groups. During 7 consecutive days, in addition to their usual diet, subjects in the soy group consumed a daily dose of 2 g of soybeans per kilogram body weight (about 170 mg/kg of isoflavones). The control group members stayed on their usual diet. The subjects were tested on mental rotation of letters and digits at the baseline and after 7 days of soy consumption. RESULTS: Consumption of soy had no influence on performance in the mental rotation task. For the intermediate stimulus rotation angle (60 deg), irrespective of soy intake, improvement of response latencies was greater in women than in men. CONCLUSION: The results indicate that soy phytoestrogens have little impact on mental rotation skill. The findings also suggest that women, in contrast to men, more readily engage memory mechanisms to solve the mental rotation task. PMID- 19300389 TI - Involvement and role of antidepressant drugs of the hypothalamic-pituitary adrenal axis and glucocorticoid receptor function. AB - Changes in the hypothalamic-pituitary-adrenal (HPA) axis are characteristic of major depression. Because the effects of glucocorticoids are mediated by intracellular receptors including, most notably, the glucocorticoid receptor (GR), several studies have examined the number and/or function of GR's in depressed patients. Review scientific evidences have consistently demonstrated that GR function is impaired in major depression, resulting in reduced GR mediated negative feedback on the HPA axis and increased production and secretion of corticotropin-releasing hormone (CRH) in various brain regions postulated to be involved in the causality of major depression. Hyperactivity of HPA axis is the main biochemical change, besides disturbed monoaminergic neurotransmission, observed in the patients suffering from a major depression. High incidence of depression in Cushing's syndrome as well as antidepressant effects of adrenocortical enzyme inhibitors in major depression support hypothesis that hyperactivity of HPA axis may be involved in pathogenesis of depression. Major alterations of the HPA axis that can be reversed by successful antidepressant therapy are often seen in depressed patients. A possible explanation for this is that the antidepressant-induced increase in GR's renders the HPA axis more sensitive to glucocorticoid feedback. This new insight into antidepressant drug action suggests a novel approach to the development of antidepressant drugs. PMID- 19300390 TI - Interhemispheric cerebral asymmetry detected by VEPS in diabetic patients with recognized depression. AB - OBJECTIVES: The present study was undertaken in order to verify the hypothesis stating that patients with depression exhibit some abnormalities concerning a cerebral symmetry. For this purpose, an analysis of the relationship between the VEPs (Visual Evoked Potentials) results and the depressive symptoms intensification, as well interrelation between depressive and diabetic symptoms were performed. MATERIAL AND METHODS: VEPs recordings were obtained from the two study groups (both aged 20-45 years), 20 healthy subjects and 32 diabetic patients with clinically documented depression. The VEPs examination was carried out using a computer system called STELLA (Stimulated Electroencephalogram on Line Analyzer). RESULTS: VEPs examinations revealed a cerebral symmetry in the all control subjects and in 10 out of 32 diabetic patients with depressive disorders. It is noteworthy that 22 of 32 diabetic patients (68.8%) showed a hemispheric asymmetry in the VEPs recordings. Of 12 patients with a moderate depression, 10 showed the left cerebral laterality, and 2--the right laterality. In turn, all 10 patients with major depressive disorder (MDD) demonstrated in the VEPs recordings a significant cerebral laterality with the right hemisphere dominance. CONCLUSIONS: The obtained results suggest that interhemispheric cerebral asymmetry might be considered a specific feature of depression, and, if this is a case, it could justify a conclusion that right hemisphere dominance could be a neurophysiological marker of MDD. It should be emphasized that intensification of the depressive symptoms has an unfavorable influence on course of diabetes mellitus, its self-control and severity of following complications. PMID- 19300391 TI - Induced abortions: still important reproduction loss in the Czech Republic? AB - OBJECTIVES: The aim of study was to evaluate the importance of induced abortions for reproduction medicine in Czech Republic. DESIGN: Demographic analysis of data published by EUROSTAT and Czech statistical office. SETTING: Department of Demography and Geodemography Faculty of Science, Charles University Prague. RESULTS: Widespread use of the liberal abortion law in socialist countries contributed to the decline of fertility rates only to the replacement level. In the Czech Republic total fertility rate dropped below 1.3 in 1995 and it did not increase above 1.5 children per woman till 2007. The increased use of modern contraceptive methods that results in a sharp decline in fertility and in a significant decrease of induced abortions can be documented. The total abortion rate fell from 1.54 abortions per a woman in 1990 to 0.34 in 2007. The proportion of women aged 15-49 years who were prescribed oral contraception increased from 4 percent in 1990 to 48 percent in 2007. An induced abortion is still used largely as a way to avoid birth of additional children by women who already have the number of children they want. This is in sharp contrast with the situation in the majority of Western European countries in which abortion is used mainly by teen age girls whose attempts to avoid pregnancy have failed. CONCLUSION: In contrast to other demographic characteristics which classify the Czech Republic to Eastern Europe, the level of induced abortion rate is comparable with the levels observed in some Western European countries. PMID- 19300392 TI - Evaluation of association between the CYP11alpha promoter pentannucleotide (TTTTA)n polymorphism and polycystic ovarian syndrome among Han Chinese women. AB - Evidence indicates that CYP11alpha is a positional and functional candidate for genetic study in polycystic ovarian syndrome (PCOS). In the present study, we have evaluated the association between the CYP11alpha promoter pentannucleotide (tttta)n polymorphism and PCOS among Han Chinese women. Subjects include 125 patients with PCOS and 121 healthy controls, and all were Han Chinese women. Clinical characteristics of patients with PCOS and control subjects were examined according to the Rotterdam consensus criteria. The CYP11alpha promoter pentannucleotide (tttta)n polymorphism was genotyped with PCR and fluorescent capillary electrophoresis protocol. Results indicate that common alleles of the CYP11alpha promoter pentannucleotide (tttta)n polymorphism in this population of Han Chinese women were P4, P6 and P8, and allele P6 was the most common one. Frequencies of those three common alleles between PCOS cases and controls (24.8, 65.6, 6.8% and 23.6, 65.7, 9.9%, respectively) were similar. No significant allelic association of this polymorphism with PCOS was found. However, the carriers with allele P6 among patients with PCOS had increased WHR (0.85+/-0.05 vs 0.82+/-0.08, P=0.039) and decreased AUCG (9.5+/-2.1 vs 11.4+/-2.8, P=0.021) compared to the patients carrying other alleles. Therefore, the most common allele of the CYP11alpha promoter pentannucleotide (tttta)n polymorphism in the population of Han Chinese women is P6, while the most common allele in European Caucasians, as previously reported, is P4. This polymorphism is an ethnic and racial variant, and may have the risk susceptibility in abnormal metabolism of patients with PCOS in Han Chinese women. PMID- 19300393 TI - Epicatechin suppresses IL-6, IL-8 and enhances IL-10 production with NF-kappaB nuclear translocation in whole blood stimulated system. AB - OBJECTIVES: Cytokines induce the balance between inflammatory versus regulatory or antibody mediated reactions. So modulating the release of cytokines or inducing them by immunomodulating agents is an attractive mode for treating or help in treating several diseases such as autoimmune diseases. Eriobotrya japonica is a plant that is traditionally thought to have anti-inflammatory activities. Several compounds were isolated from the plant and showed distinctive biological effects. The purpose of this study was to determine the effects of epicatechin (EC) isolated from Eriobotrya Japonica on IL-6, IL-8, and IL-10 productions in whole blood stimulated with phytohemagglutinin (PHA)+lipopolysaccharide (LPS), and to examine if these cytokines are modulated through NFkappaB pathway. METHODS: Sixteen healthy males and females volunteered in the study. Blood samples were drawn, diluted, and cultured for 24 h with different concentrations of EC and then PHA+LPS was added for another 24 h. The supernatant, then, was harvested and assayed for cytokines. In addition, mixing studies of EC and hydrocortisone were performed to examine the cytoplasmic and nuclear fractions of NFkB levels in association with cytokine production levels. RESULTS: Increasing concentrations of EC (1-100 microg/ml) in PHA+LPS stimulated whole blood cells culture suppressed significantly (p<0.001) the production of IL 6 and IL-8. Moreover, increasing concentrations of EC modulated significantly the production of IL-10, as there was a significant increase in IL-10 level at 0.1, 1.0, and 10 microg/ml (p=0.058-0.004), while a significant decrease at a concentration of 100 microg/ml EC (p=0.037) was observed. In addition, an additive effect between EC and hydrocortisone (HC, 100 nmol/l) was seen in the production of IL-10, as there was a significant increase in IL-10 level (32%) compared with 27% for EC (10 microg/ml) and 19% HC. Furthermore, a significant decrease in cytoplasmic fractions of NFkappaB p65 level was found in samples containing EC 1, 10 microg/ml, but not in 100 microg/ml, when compared with control (p<0.03). These latter changes were accompanied with a 29%, 67%, and 98% increase, respectively, of NFkappaB p65 in nuclear fractions compared to 24% reduction of NFkappaB p65 level in HC culture (p<0.03). CONCLUSIONS: These results indicate that EC suppresses the production of pro-inflammatory cytokines, IL6 and IL-8, enhances the production of anti-inflammatory cytokine, IL-10, and stimulates NFkappaB p65 translocation to nucleus in PHA+LPS stimulated whole blood culture. PMID- 19300394 TI - Enhancement of oral bioavailability of insulin in humans. AB - OBJECTIVE: The purpose of this study is to investigate oral absorption of 1, 2 and 3 U/kg oral insulin five test products with different particle sizes in comparison with 0.1 U/kg subcutaneous reference formulation. METHODS: Twenty five healthy volunteers participated in five studies using a two-phase, two-sequence crossover design with washout period of one day. Mean disposition kinetics was determined by non-compartmental analysis using Kinetica program. Absorption kinetics of insulin products were then determined using SIMCYP simulator utilizing ADAM model. RESULTS & CONCLUSIONS: Dimensional analysis results showed the superiority of formula 4:2 U/kg oral dose with 57 nm particle size over other oral formulations when compared with subcutaneous route. Optimized intestinal permeability coefficients (x10(-4)) of insulin best test and reference formulations were 0.084 and 0.179 cm/sec respectively. Total fraction of insulin dose absorbed (Fa) for the test and reference products were 3.0% and 19% respectively. Subcutaneous product exhibited higher absorption rate and extent than oral insulin. Yet that was compensated by the increase in other factors such as Fa*, Peff* and oral dose, leading to similar insulin plasma levels and similar effect on glucose infusion rates. Oral insulin bioavailability was shown promising for the development of oral insulin product. PMID- 19300395 TI - Effects of rosiglitazone--peroxisome proliferators-activated receptor gamma (PPARgamma) agonist on cell viability of human pituitary adenomas in vitro. AB - OBJECTIVES: Rosiglitazone (RGZ) belongs to thiazolidinediones - new class of antidiabetic drugs which are PPARgamma agonists. It was shown that tumoral tissue, including the pituitary adenomas, posses PPARgamma receptors. The activation of PPARgamma receptors inhibits tumour growth in rodents and induces the oncostatic effect on human cancer cell lines. The aim of the present study was to examine the anti-tumour effect of RGZ on human pituitary adenomas in vitro. MATERIALS AND METHODS: Cells of eight pituitary adenomas removed neurosurgically were used to our experiment. Before the operation, the hormonal secretion of the tumour was estimated. After the surgery, the histological diagnosis and immunohistochemical detection of pituitary hormones and PPARgamma receptors were performed. The cells of pituitary tumours were exposed in the primary culture to RGZ at the concentrations of 10(-9)-10(-4) M for 24 hours. To measure the cell growth the modified colorimetric Mossman method detecting the cells viability was applied. RESULTS: On the basis of the pre-operative diagnosis the 6 clinically non-functioning adenomas (CNFPA), one case of acromegaly and one case of Cushing's disease were recognized. In 5 out of 6 CNFPA the immunopositive reaction for different pituitary hormones such as: LH, HGH, PRL, FSH and alpha subunit was detected. Expression of PPARgamma was found in all examined tumours. Rosiglitazone decreased the cell viability of all CNFPA and corticotropinoma for 20% or more. In somatotropinoma inhibition of the cell growth was about 13%. There is no correlation between PPARgamma expression and efficacy of rosiglitazone. THE MAIN FINDING: The obtained results indicate that RZG exerts a suppressive effect on the cell viability in non-functioning pituitary adenomas. The lack of correlation between PPARgamma expression and anti-tumoral effect of RZG suggests that the above-mentioned action of this compound is independent on PPARgamma expression. CONCLUSION: Our data suggest that rosiglitazone may be useful in the treatment of non-functioning pituitary adenomas, but its efficacy in Cushing's disease and acromegaly requires further study. PMID- 19300396 TI - Differential binding of mammalian and salmon GnRHs with rat and carp pituitary receptors. AB - OBJECTIVES: Receptor binding of GnRH is connected with the stimulation of pituitary gonadotropic cells leading to both the release and biosynthesis of gonadotropins. The binding is connected with the conformational changes in the receptor which induce the specific intracellular signalisation. The study of fish GnRHs and their receptors may give us new knowledge of the complex interplay of different mechanisms involved in neuroendocrine regulation of reproduction. METHODS: Receptor binding of both mGnRH and sGnRH were compared by the study utilizing the displacement method with mGnRH or sGnRH as radioactive tracers. Incubation was performed at 2 degrees C to avoid ligand degradation. RESULTS: The comparative binding of mGnRH and sGnRH with GnRH receptors from the female rat pituitary and female carp pituitary was studied. At the 50% of displacement, the binding of sGnRH to the rat pituitary receptor was very small and in comparison to the binding of mGnRH (100%) was in the range 2-15%. However, the binding of mGnRH to carp pituitary receptors is small in comparison with the binding of sGnRH (100%) and was in the range 5-20%. CONCLUSION: The results demonstrated the differences in binding of different GnRHs to the receptor in rats and carp. This suggests that the structures of GnRH and its receptor undergo co-evolution in different classes of animals. PMID- 19300397 TI - Differences in the blood serum levels of soluble HLA-G concentrations between the menstrual cycle phases and menopause in patients with ovarian endometriosis and uterine leiomyoma. AB - BACKGROUND: HLA-G is an antigen whose participation in the regulation of the immune system is well documented. The aim of the present study has therefore been to evaluate the sHLA-G blood serum concentrations levels in both women with ovarian endometriosis and women with uterine leiomyomas. METHODS: In our study, the soluble HLA-G concentration level was evaluated in the blood serum samples obtained from 98 women who underwent laparotomies or laparoscopies due to either ovarian endometriosis or leiomyomatous uterus. The control group consisted of 42 women, including women on whom a diagnostic laparoscopy identified no lesions, and volunteers-healthy women who returned their blood serum samples during menstrual bleeding. RESULTS: Patients who underwent surgical treatment because of ovarian endometriosis or uterine leiomyoma, as well as patients from the control group, exhibited no sHLA-G blood serum concentration level fluctuations between the proliferative and secretory menstrual cycle phases. The sHLA-G levels were significantly lower in the patients with ovarian endometriosis and in the patients from the control group during the menstrual cycle phase than in those patients with leiomyoma. A similar relation between the sHLA-G levels of the postmenopausal patients suffering from leiomyoma and the control patients was found. In contrast, the postmenopausal women suffering from endometriosis were typified by levels of sHLA-G blood serum concentration comparable to those of the patients with leiomyoma, and the levels were significantly higher than those observed in the blood sera of the postmenopausal patients from the control group. CONCLUSION: The soluble HLA-G blood serum level would seem to be a useful marker for evaluating the status of the microenvironment, where the tumor-immune cell and ectopic and eutopic endometrial interactions take place. PMID- 19300398 TI - Correlation amyloid in brain, kidney, and CSF of castrated guinea pigs. AB - OBJECTIVES: The association between low testosterone levels and Alzheimer's disease (AD) amyloid beta-peptide (Abeta) metabolism was investigated in brain and kidney of guinea pigs. METHODS: The expression of Abeta peptide in the brain and kidney was assessed by using the immunohistochemistry method. RESULTS: No expression of Abeta was seen in both groups of animals. This negative staining was found until the fourth week following castration. The formation of Abeta in guinea pigs is perhaps not a short duration process and may undergo different metabolic pathway compare to humans. CONCLUSION: castration was not associated with the formation of Abeta in the brain and kidneys during a 1-month period and might require a longer period of time. PMID- 19300399 TI - Combination of voxel based morphometry and diffusion tensor imaging in patients with Alzheimer's disease. AB - OBJECTIVE: The aim of the study was to assess structural changes in gray matter (GM) volume and fractional anisotropy (FA) in patients with Alzheimer's disease (AD) compared to control subjects using Voxel-Based Morphometry (VBM). Fractional anisotropy in the corpus callosum of both groups was also calculated using ROI analysis. METHODS: Twenty-one patients and twenty-three control subjects underwent MRI examination using T1-weighted 3D MPRAGE sequence and diffusion spin echo echo-planar imaging sequence in six directions. Structural MRI analyses for GM volume and FA were performed using an optimized VBM protocol implemented in SPM5. The influence of age and Mini-Mental State Examination (MMSE) was dealt with multiple regression analysis either for the whole group or for AD patients and controls separately. RESULTS: Patients showed significant reduction of GM volume mainly in the temporal lobes. In AD patients, no correlation was observed between GM volume and age or MMSE. FA was reduced in AD patients mainly in frontal and temporal lobes. In both groups no correlation was found between FA and age or MMSE. Patients with AD showed a significant decrease in FA and an increase in mean diffusivity (p<0.0001) in the corpus callosum. CONCLUSIONS: In patients with AD we observed a significant reduction in FA values and GM volume; however, no correlation with age and MMSE was proven for both FA and GM for AD patients. This finding supports the hypothesis that morphological changes in patients with AD are not a continuous aging related process but represent qualitative changes. PMID- 19300400 TI - Cerebrospinal fluid levels of iodothyronines and nerve growth factor in patients with multiple sclerosis and neuromyelitis optica. AB - OBJECTIVE: Multiple sclerosis (MS) is an inflammatory demyelinating disease of the human central nervous system (CNS) and a major cause of neurological disability among adults in North America and Europe. Neuromyelitis optica (NMO) is a very severe disease of inflammatory demyelination located in the optic chiasm, nerves and the spinal cord. The aim of this study is to assess thyroid hormone (TH) and nerve growth factor (NGF) levels in cerebrospinal fluid (CSF) of MS, NMO patients and controls, and investigate whether there is any correlation between TH and NGF levels in the CSF. PATIENTS AND METHODS: 38 relapsing remitting multiple sclerosis (RRMS), 10 NMO and 19 controls were investigated whether there was any correlation between TH and NGF levels in the CSF. RESULTS: MS and NMO patients exhibited significantly higher CSF NGF (respectively P<0.05, P<0.05), TT4 levels (P<0.001) and higher TT4/ rT3 ratio (respectively P<0.01, P<0.01) compared with the controls. Significant correlation was found between CSF NGF levels and CSF rT3 levels or TT4/ rT3 ratio in controls (respectively P<0.01, P<0.05). EDSS was significantly correlated with CSF rT3 levels and TT4/ rT3 ratio in MS patients (respectively P<0.05, P<0.001). CONCLUSIONS: These results indicate that an abnormal thyroid hormone may exist within the brain in the patients with MS. CSF rT3 levels and TT4/ rT3 ratio could be regarded as useful markers of underlying disease activity. PMID- 19300401 TI - Brain levels of GABA, glutamate and aspartate in sociable, aggressive and timid mice: an in vivo microdialysis study. AB - OBJECTIVES: Some individually-housed male mice behave aggressively during encounters with strange males, while others are timid or sociable in the same situation. The objective of the present study was to examine concentrations of glutamate, aspartate, and GABA in the brain of aggressive, timid, and sociable mice. METHODS: Random-bred albino mice were housed individually for three weeks and then classified in three groups (aggressive, timid, and sociable mice) according to their behavior during social interaction with non-aggressive group housed male mice in a neutral cage. One week after categorization, by means of the social conflict test, levels of glutamate, aspartate, and GABA were measured by in vivo microdialysis of the medial prefrontal cortex (mPFC) of the isolated and group-housed mice. RESULTS: Sociable mice had almost triple the levels of GABA in their mPFC than aggressive or timid mice. No significant differences in aspartate and glutamate levels were found in these three types of individually housed mice. Forebrain chemistry of group-housed mice did not differ from that of individually-housed mice with the exception of levels of glutamate and GABA which were significantly lower in group-housed mice than in sociable individually housed mice. CONCLUSION: The present results suggest that GABA might play a role in sociable behavior. Results also corroborate other findings indicating that the GABAergic system represents an important molecular and neuronal substrate for the selective attenuation of anxiety and aggression. PMID- 19300402 TI - Bradykinin receptors antagonists and nitric oxide synthase inhibitors in vincristine and streptozotocin induced hyperalgesia in chemotherapy and diabetic neuropathy rat model. AB - PURPOSE: The influence of an irreversible inhibitor of constitutive NO synthase (L-NOArg; 1.0 mg/kg ip), a relatively selective inhibitor of inducible NO synthase (L-NIL; 1.0 mg/kg ip) and a relatively specific inhibitor of neuronal NO synthase (7-NI; 0.1 mg/kg ip), on antihyperalgesic action of selective antagonists of B2 and B1 receptors: D-Arg-[Hyp3,Thi5,D-Tic7,Oic8] bradykinin (HOE 140; 70 nmol/kg ip) or des Arg10 HOE 140 (70 nmol/kg ip) respectively, in model of diabetic (streptozotocin-induced) and toxic (vincristine-induced) neuropathy was investigated. METHODS: The changes in pain thresholds were determined using mechanical stimuli--the modification of the classic paw withdrawal test described by Randall-Selitto. RESULTS: The results of this paper confirm that inhibition of bradykinin receptors and inducible NO synthase but not neuronal NO synthase activity reduces diabetic hyperalgesia. Pretreatment with L-NOArg and L-NIL but not 7-NI, significantly increases antihyperalgesic activity both HOE 140 and des Arg10 HOE 140. It was also shown that both products of inducible NO synthase and neuronal NO synthase activation as well as bradykinin are involved in hyperalgesia produced by vincristine. Moreover, L-NOArg and 7-NI but not L-NIL intensify antihyperalgesic activity of HOE 140 or des-Arg10HOE 140 in toxic neuropathy. CONCLUSIONS: Results of these studies suggest that B1 and B2 receptors are engaged in transmission of nociceptive stimuli in both diabetic and toxic neuropathy. In streptozotocin-induced hyperalgesia, inducible NO synthase participates in pronociceptive activity of bradykinin, whereas in vincristine induced hyperalgesia bradykinin seemed to activate neuronal NO synthase pathway. Therefore, concomitant administration of small doses of bradykinin receptor antagonists and NO synthase inhibitors can be effective in alleviation of neuropathic pain, even in hospital care. PMID- 19300403 TI - Bombesin modulates the control of energy homeostasis and pituitary hormone release. AB - Bombesin and related peptides are widely distributed in gastrointestinal tract and central nervous system. It has been reported that they play an important role in the control of appetite, metabolism, sensory transmission and thermoregulation as well as in the regulation of pituitary hormone release. Central injection of these peptides leads to inhibition of feeding. There are controversial opinions about the effects of bombesin on pituitary hormone secretion both in vivo and in vitro experiments. PMID- 19300405 TI - Analysis of treatment efficacy in girls with anorexia nervosa (III). AB - OBJECTIVES: So far, no unequivocal answer has been provided to the question of how to treat girls with anorexia nervosa (AN). The goal of the study was evaluation of treatment results and search for prognostic factors, regarding the outcome of administered therapy in girls with AN. PATIENTS AND METHODS: The analysis comprised eighty-seven (87) patients with diagnosis of AN. In the course of treatment, the patients received only a balanced and highly energetic diet. The effects of dietetic procedure were evaluated, comparing clinical data and routine laboratory test results (full blood cell count, serum concentrations of sodium, potassium, glucose, cholesterol, triglycerides, total calcium, phosphates, total protein and the urea) at the onset and on the last day of hospitalisation and also an analysis of clinical data in patients with regained menstrual cycles was performed. Also, on admission to hospital, serum concentrations of TSH, FT4, FT3, E2, T and cortisol were measured (the last hormone - three times - at 8:00, 17:00 and 24:00). Moreover, a stimulation test with GnRH was performed to assess LH and FSH release. A comparison was made between the clinical data, the results of routine laboratory tests and hormone concentrations, obtained at the beginning of the therapy in the group of cured girls (the girls with regained menses) and the group, in which the therapy did not bring complete elimination of the disease. RESULTS: After hospitalisation, 75.9% of the patients with AN continued outpatient treatment. In all the patients, who adhered to proposed therapy, menstruation cycles returned within 1 to 12 months. It was found that in the group with positive treatment outcome, the patients were older, demonstrated higher BMI values at the beginning of the therapy, as well as shorter disease duration than those in the group of patients who refused further treatment. In the group of cured patients, estradiol concentrations were significantly higher and cortisol concentrations (at 17:00 and 24:00) were significantly lower, in comparison to the uncured group. CONCLUSIONS: The findings suggest that body weight increase by proper diet application is the most important element of therapy in AN. Patient's age, disease duration, the degree of body weight loss and serum concentrations of estradiol and cortisol at 17:00 and 24:00 are the prognostic factors for the outcome of therapy in anorexia nervosa. PMID- 19300406 TI - Trends in tuberculosis--United States, 2008. AB - In 2008, a total of 12,898 incident tuberculosis (TB) cases were reported in the United States; the TB rate declined 3.8% from 2007 to 4.2 cases per 100,000 population, the lowest rate recorded since national reporting began in 1953. This report summarizes provisional 2008 data from the National TB Surveillance System and describes trends since 1993. Despite this overall improvement, progress has slowed in recent years; the average annual percentage decline in the TB rate decreased from 7.3% per year during 1993-2000 to 3.8% during 2000-2008. Foreign born persons and racial/ethnic minorities continued to bear a disproportionate burden of TB disease in the United States. In 2008, the TB rate in foreign-born persons in the United States was 10 times higher than in U.S.-born persons. TB rates among Hispanics and blacks were nearly eight times higher than among non Hispanic whites, and rates among Asians were nearly 23 times higher than among non-Hispanic whites. In 2008, among persons with TB whose country of origin was known, approximately 95% of Asians, 76% of Hispanics, 32% of blacks, and 18% of whites were foreign born. Among U.S.-born racial and ethnic groups, the greatest racial disparity in TB rates was for U.S.-born blacks, whose rate was seven times higher than the rate for U.S.-born whites. Intensified efforts are needed to address the slowing decline in TB incidence and the persistent disparities that exist between U.S.-born and foreign-born persons and between whites and minorities in the United States. PMID- 19300407 TI - Two simultaneous outbreaks of multidrug-resistant tuberculosis--Federated States of Micronesia, 2007-2009. AB - In July 2008, CDC responded to a request from the Federated States of Micronesia (FSM) to investigate the first documented cases of multidrug-resistant tuberculosis (MDR TB) in Chuuk State. Compared with drug-susceptible TB disease, MDR TB is resistant to at least isoniazid and rifampin, the two most effective TB medications, making treatment more difficult and outcomes more likely fatal. Second-line TB drugs for treating MDR TB were not available in FSM, and during December 2007-June 2008 four patients with MDR TB had died, including a child aged 2 years. This report describes the investigation by the World Health Organization (WHO) and CDC, which initially identified five confirmed cases in two distinct clusters, characterized by two distinct geographic locations, genotypes, and drug-susceptibility patterns. Extensive transmission has occurred among household contacts; 16 (8%) of the 205 contacts identified have confirmed or suspected MDR TB disease, and 124 (60%) have latent TB infection. Among 21 confirmed and suspected cases of MDR TB identified as of March 13, 2009, 10 have been in persons aged <15 years. With the death of a child aged 4 years in November 2008, a total of five persons have died of MDR TB. Multiple U.S. government agencies and other organizations are assisting local health authorities with resources to procure second-line TB drugs, ensure directly observed therapy (DOT), and identify and evaluate contacts. These simultaneous and continuing outbreaks demonstrate how a lack of basic TB control activities can allow the emergence and spread of drug-resistant TB. PMID- 19300408 TI - Guidance for control of infections with carbapenem-resistant or carbapenemase producing Enterobacteriaceae in acute care facilities. AB - Infection with carbapenem-resistant Enterobacteriaceae (CRE) or carbapenemase producing Enterobacteriaceae is emerging as an important challenge in health-care settings. Currently, carbapenem-resistant Klebsiella pneumoniae (CRKP) is the species of CRE most commonly encountered in the United States. CRKP is resistant to almost all available antimicrobial agents, and infections with CRKP have been associated with high rates of morbidity and mortality, particularly among persons with prolonged hospitalization and those who are critically ill and exposed to invasive devices (e.g., ventilators or central venous catheters). This report provides updated recommendations from CDC and the Healthcare Infection Control Practices Advisory Committee (HICPAC) for the control of CRE or carbapenemase producing Enterobacteriaceae in acute care (inpatient) facilities. For all acute care facilities, CDC and HICPAC recommend an aggressive infection control strategy, including managing all patients with CRE using contact precautions and implementing Clinical and Laboratory Standards Institute (CLSI) guidelines for detection of carbapenemase production. In areas where CRE are not endemic, acute care facilities should 1) review microbiology records for the preceding 6-12 months to determine whether CRE have been recovered at the facility, 2) if the review finds previously unrecognized CRE, perform a point prevalence culture survey in high-risk units to look for other cases of CRE, and 3) perform active surveillance cultures of patients with epidemiologic links to persons from whom CRE have been recovered. In areas where CRE are endemic, an increased likelihood exists for imporation of CRE, and facilities should consider additional strategies to reduce rates of CRE. Acute care facilities should review these recommendations and implement appropriate strategies to limit the spread of these pathogens. PMID- 19300409 TI - Amplification and co-regulators of androgen receptor gene in prostate cancer. AB - Prostate cancer is the second most common malignancy among males after lung cancer. The growth of prostate cancer cells depends on the presence of androgens, a group of steroid hormones that include testosterone and its more active metabolite dihydrotestoste-rone. Most prostate cancers are androgen-dependent and respond to the antiandrogens or androgen-deprivation therapy. However, the progression to an androgen-independent stage occurs frequently. Possible mechanisms that could be involved in the development of hormone resistant prostate cancer causes including androgen receptor (AR) mutations, AR amplification/over expression, interaction between AR and other growth factors, and enhanced signaling in a ligand-independent manner are discussed. PMID- 19300410 TI - Photochemoprevention of cutaneous neoplasia through natural products. AB - Non-melanoma skin cancers such as squamous cell carcinoma and basal cell carcinoma are the most common types of human tumors, representing 30% of the new cases of malignancies diagnosed each year. Ultraviolet radiation (UV) from the sun is a major cause of non-melanoma skin cancer in humans. The prevention and mainly the photochemoprevention with natural products represent a simple but very effective strategy in the management of cutaneous neoplasia. Here we review the progress in the research of new and existing agents developed to protect the skin exposed to UV. We also discuss the current state of knowledge on their photosuppression mechanism in humans as well as in animal models, and efficiency in cancer prevention. PMID- 19300411 TI - Effects of low and high concentrations of antitumour drug taxol in anaplastic thyroid cancer cells. AB - AIM: To study the changes of cell cycle, mitochondrial membrane potential and caspase activation in response to an antitumour drug Taxol in ARO and KTC-2 cell lines of anaplastic thyroid carcinoma. METHODS: Experiments were done on thyroid anaplastic cancer cell lines ARO and KTC-2 using Western blotting, flow cytometry, light and fluorescent microscopy. RESULTS: Taxol significantly activated caspases in ARO cells starting from drug concentration of 5 nM. Maximum activation was observed at 25 nM and further increase of Taxol concentration to 100 nM resulted in a reduction of caspase activation. Concomitant to caspase activation, a loss of mitochondrial membrane potential was observed. At Taxol concentration of 100 nM, most cells lost their mitochondrial membrane potential. Low Taxol concentrations (10 nM) caused changes in the cell cycle that are typical for apoptosis without cell cycle arrest. Higher drug doses starting from 50 nM arrested cell cycle in G2/M phase. In KTC-2 cell line Taxol concentration as low as 1 nM induced apoptosis. 6-15 nM of the drug caused massive (75-83%) cell death. Upon Taxol action, the increase in the number of cells displaying manifestations of accelerated senescence was insignificant. CONCLUSION: Taxol induces bona fide apoptosis in thyroid cancer cell cultures at low (1-25 nM) concentrations. Higher drug doses cause the loss of mitochondrial membrane potential and possibly lead to other types of cell death. No accelerated senescence at different Taxol concentrations was observed. The significance of subG1 and G2/M cell populations at low and high doses of Taxol is discussed. PMID- 19300412 TI - Synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor agent. AB - AIM: To develop a rationally designed new organotin compound namely dimethyl tin 4-cyclohexyl thiosemicarbazone (D4-t) and evaluate its putative antitumor activity. METHODS: Starting from 4-cyclohexyl thiosemicarbazone, a three step synthetic procedure was followed to obtain the title compound. In vivo lymphocyte activation property of the compound at three different doses was assayed by measuring the blastogenesis. Concanavalin A (ConA) was used as standard mitogen for murine T cells stimulation in vivo . Also, the synthesis of DNA by the activated lymphocytes was measured after injecting the D4-t. The lymphocyte activation property and antitumor efficacy of D4-t were assessed in Sarcoma-180 (S-180) bearing mice. The organization of lymphoid cells was studied in the histological preparations of spleen and mesenteric lymph node. Tumor neutralization assay (Winn assay) was conducted to examine whether immune responses were associated with the manifestation of antitumor efficacies of this compound in S-180 in vivo . The DNA synthesis inhibitory effect of the compound in S-180 cells was studied in vitro, and was found significant (P < 0.001). RESULTS: Different doses of the new compound caused differential response of blastogenesis and DNA synthesis. In comparison to ConA, the title compound showed a good number of blast cells at its optimum dose of 5 mg/kg. It caused maximum synthesis of DNA by the lymphoid cells. In histological preparations, the gradual transformation of lymphocytes into blasts was observed without any visible toxicity. Winn assay revealed that 5 mg/kg of D4-t was able to reduce tumor mass without severe toxicity. This organotin compound also inhibits the synthesis of DNA in S-180 tumor cells in comparison to Platin10 and ConA. CONCLUSION: The title compound has the lymphocyte activation property and stimulates immune response of the lymphoid cells, which in turn express the antitumor activity without any significant toxicity. Results indicate promising therapeutic potential of D4-t. PMID- 19300413 TI - In vitro and in vivo anticancer activity of steroid saponins of Paris polyphylla var. yunnanensis. AB - AIM: To confirm the anticancer activity of steroid saponins isolated from the rhizome of Paris polyphylla var. yunnanensis and evaluate the structure-activity relationships of these steroid saponins in vitro and in vivo . MATERIALS AND METHODS: Eight known steroid saponins were isolated from the rhizome of Paris polyphylla . The LA795 lung adenocarcinoma cell line from mice was chosen to evaluate cytotoxicity by means of MTT assay, and to study apoptosis by means of AnnexinV-FITC/PI flow cytometry. Diosgenin-3alpha-L-arabinofuranosyl(1-->4) [alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glycopyranoside (compound 1), the main steroid saponin of Paris polyphylla , and diosgenin (Dio), the aglycone of major steroid saponins, were evaluated for antitumor activity on LA795 lung adenocarcinoma in T739 inbred mice. RESULTS: The steroid saponins showed remarkable cytotoxicity and caused typical apoptosis in a dose-dependent manner. They were evaluated in vivo by their effect on tumor developed in T739 inbred mice. The oral administration to T739 mice bearing LA795 lung adenocarcinoma of compound 1 and diosgenin significantly inhibited tumor growth, by 29.44% and 33.94%, respectively. HE staining showed that lungs and livers of treated mice underwent various levels of histopathological alterations. It was demonstrated by TUNEL assay that apoptosis rate in tumor cells was increased in comparison to cells in control mice. The 3-O-glycoside moiety and spirostanol structure played an important role in the anticancer activity of steroid saponins, and the number and the variety of glycosides of compounds strongly influenced on their anticancer activity. CONCLUSION: Rhizoma Paridis saponins showed anticancer activity against lung adenocarcinoma cell line, both in vitro and in vivo, and their effect was dependent on compounds' structure in a certain degree. PMID- 19300414 TI - Influence of two Pt(IV) complexes on viability, apoptosis and cell cycle of B16 mouse melanoma tumors. AB - Several platinum(IV) complexes are showing considerable promise in initial trials, producing reactive intermediates that then interact with DNA. AIM: To perform in vitro study of two new platinum(IV) complexes cytotoxic effect on B16 mouse melanoma cells. METHODS: PtCl(4)(dbtp)(2) and PtCl(2)(6mp)(2) complexes were prepared. PtCl(4)(dbtp)(2) was created as modification of PtCl(4)(dmtp) test previously. Apoptosis and necrosis were examined using flow cytometry, upon Annexin V/PI staining. RESULTS: LC(10), LC(50) and LC(90) parameters established for PtCl(4)(dbtp)(2) were as following: 2.6, 17.0, 58.0 mumol/L. However LC(10) and LC(50) established for PtCl(2)(6mp)(2) were 1.2 and 14.0 micromol/l respectively. The both complexes induced apoptosis. PtCl(2)(6mp)(2) induced cell cycle arrest in G0/G1, while PtCl(4)(dbtp)(2) - in S-phase. CONCLUSIONS: PtCl(4)(dbtp)(2) appeared to be more cytotoxic against B16 cells than PtCl(2)(6mp)(2). Apoptosis was the main mechanism of cell loss in cultures incubated with both tested complexes. PMID- 19300415 TI - The study of phosphate transporter NAPI2B expression in different histological types of epithelial ovarian cancer. AB - The identification of markers that are specifically expressed by different histological types of epithelial ovarian cancer (EOC) may lead to the development of novel and more specific diagnostic and therapeutic strategies. Sodium dependent phosphate transporter NaPi2b (or MX35 ovarian cancer antigen) is a novel perspective marker of EOC. To date, the studies on NaPi2b/MX35 expression in different histological types of EOC are limited. AIM: To examine NaPi2b/MX35 expression in different histological types of epithelial ovarian tumors. METHODS: Here, we describe the analysis of NaPi2b expression in serous (n = 17), endometrioid (n = 8), and mucinous ovarian tumors (n = 3) by Western-blotting (WB), immunohistochemistry and RT-PCR. RESULTS: The results of immunohistochemical and WB analysis showed that benign and well-differentiated malignant papillary serous tumors as well as well-differentiated malignant endometriod tumors overexpress NaPi2b protein. However, no overexpression of NaPi2b was detected in benign and malignant mucinous tumors as well as in poorly differentiated endometriod tumors. Notably, the expression NaPi2b mRNA was detected in all investigated histological types of EOC. CONCLUSION: We have shown the differential expression profile of NaPi2b phosphate transporter at protein level in various histological types of epithelial ovarian cancer. This finding might facilitate the development of more effective approaches for diagnosis and treatment of this disease. PMID- 19300416 TI - Quantitative detection of cytokeratin 20 mRNA in urine samples as diagnostic tools for bladder cancer by real-time PCR. AB - AIM: To determine and compare cytokeratin 20 (CK20) mRNA expression in urine of patients with transitional cell carcinomas of bladder (TCCB), urological benign diseases, and healthy volunteers. METHODS: Taqman probe was designed according to the sequence of CK20 cloned gene. The quantitative PCR reaction system was optimized and evaluated. The CK20 mRNA level was screened by real-time polymerase chain reaction (RT-PCR) in 95 urine samples and analyzed according to the following parameters: urinary cytology, nuclear matrix protein 22 (NMP22) expression, tumor stage and grade. RESULTS: For 60 TCCB patients urinary cytology was positive in 28 (46.7%), control group had no false-positive results (specificity 100%). CK20 expression was positive in RT-PCR of 51 cases (85%) of TCCB, but control group was positive in 2 cases (specificity 94.3%) with a cutoff value of crossover point (CT) = 30. Two methods have significant variation in sensitivity (p < 0.001), NMP22 expression was positive in 47 cases (78.3%), but control group was positive in 9 cases (specificity 85%). In the simultaneous evaluation of CK20 and NMP22 mRNA expression, there were 54 positive cases (90%). CK20 mRNA values in TCCB group (mean 27712.57 copies/microl) were significantly higher than in non-cancer disease urological group (mean 74.45 copies/microl) and control group (mean 8.47 copies/microl) (p < 0.001, p < 0.001, respectively). CK20 mRNA values increased gradually with higher tumor grade and stage: G1 differs significantly from G2 (p = 0.016); T(is) /T(a) differs significantly from T(1-2) (p = 0.022). CONCLUSION: Our results indicate that CK20 mRNA expression could be regarded as a potential marker for TCCB. We demonstrated correlation between CK20 expression and the clinicopathologic features of TCCB (tumor stage and grade); simultaneous use of CK20 and NMP22 markers will elevate the sensitivity of the method. CK20 RT-PCR is a sensitive, quantitative, rapid and specific method to detect free cancer cells in the urine, and could be recommended for be wide application in the diagnostics of TCCB and evaluation of therapeutic effect. PMID- 19300417 TI - Expression of the estrogen and progesterone receptors as prognostic factor in serous ovarian cancers. AB - AIM: To study the expression of estrogen receptors (ER) and progesterone receptors (PR) and proliferation marker Ki-67 in ovarian tumors using immunohistochemistry, and evaluate possible prognostic significance of these markers. METHODS: Immunohistochemical evaluation of Ki-67, ER and PR expression was performed on serous ovarian cancer (OC) tissue samples from 81 OC patients. RESULTS: Serous OC is characterised by high proliferative activity and increased expression of steroid hormone receptors compared to nontransfomed ovarian surface epithelium. It has been shown that ER and PR expression levels depend on tumor histologic grade and the stage of the disease, and are variable between tumors of the same grade. The ER and PR expression levels correlate with OC patients' survival. CONCLUSION: Proliferative activity and steroid hormone receptor status along with clinical and morphological characteristics of serous OC possess prognostic significance and may be used for evaluation of the disease course. PMID- 19300418 TI - Even a mild anemia is related to tumor aggressiveness mediated by angiogenic factors. AB - Esophagogastric cancers have high recurrence rates with lymph nodes being a common pattern. Pre-treatment anemia has been reported an independent prognostic factor of treatment failure regardless of treatment strategy, particularly associated with poor locoregional control. A causative relationship between anemia - tumor hypoxia - tumor aggressiveness mediated by angiogenesis up regulation is advocated, yet remains controversial. AIM: To determine whether and how the pre-treatment anemia is associa-ted with various aspects of disease aggressiveness and to evaluate the possible involvement of angiogenesis mediators. METHODS: In 111 esophagogastric cancer patients we investigated the association of pre-treatment hemoglobin concentration and anemia presence with cancer-related, patients-related features and laboratory parameters including angiogenic factors: vascular endothelial growth factors A and C, interleukin-8 and midkine. Serum levels of angiogenic factors were assessed with immunoenzymatic tests. RESULTS: Histology, disease stage, regional metastasis and dissemination in general, malnutrition and angiogenesis represented by midkine were found to correlate with anemia presence and hemoglobin concentration, while tumor extension, patient's age and sex accounted only for anemia presence. A tendency towards hemoglobin correlation with VEGF-A and Il-8 was also observed. Midkine, tumor histology and malnutrition were found to exert an independent effect on pre-treatment hemoglobin concentration and anemia presence in esophagogastric cancer patients. Hemoglobin level of 12 g/dL was found an optimal cut-off value for discrimination between localized and disseminated cancers. CONCLUSIONS: Even a mild pre-treatment anemia is associated with cancers metastasizing especially to regional lymph nodes, which seems to be mediated by some of studied angiogenic factors. PMID- 19300419 TI - Polymorphisms of the DNA base excision repair gene MUTYH in head and neck cancer. AB - BACKGROUND: Head and neck squamous cell carcinomas (HNSCC) comprise about 6% of all malignant neoplasms. The major risk factors of -HNSCC are smoking and alcohol consumption. Genetic polymorphisms of DNA repair enzymes may lead to genetic instability and carcinogenesis. MUTYH gene encodes a DNA glycosylase that can initiate the base excision repair (BER) pathway and prevent G:C > T:A transversion by excising adenine mispaired with 8-hydroxyguanine produced by reactive oxygen species (ROS). AIM: to perform a case-control study to test the association between polymorphism in the MUTYH gene: Tyr165Cys and head and neck cancer risk progression. METHODS: Genotypes were determined in DNA from peripheral blood lymphocytes of 193 patients (among them 97 subjects with precancerous hyperplastic laryngeal lesions and 96 subjects with head and neck cancer) and 140 age, sex and ethnic-matched cancer-free controls by tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR). RESULTS: We found an association between head and neck cancer risk and the Tyr165Tyr variant of the MUTYH gene (OR 2.18; 95% CI 1.19-3.97). For Tyr165Tyr genotype we also observed positive correlation with cancer progression assessed by tumor size (OR 4.56; 95% CI 1.60-12.95). We did not observe any correlation between Tyr165Cys polymorphism of MUTYH gene and precancerous hyperplastic laryngeal lesions risk. CONCLUSION: The Tyr165Tyr polymorphic variant of the MUTYH gene may be associated with head and neck cancer in Polish population. PMID- 19300420 TI - Tyrosine kinase inhibitors in treatment of fibrous histiocytoma. AB - AIM: To describe potential beneficial effects of tyrosine kinase inhibitor in the treatment of unresectable/metastatic fibrous histiocytoma. METHODS: We report a case of advanced stage fibrous histiocytoma with locally recurrent disease plus lung and bone metastatic deposits. Patient was treated with the tyrosine kinase inhibitor sunitinib. RESULTS: Treatment with Sunitinib resulted in disease stabilization in the regional lesion and in good partial response for metastatic foci (reduction in number and size). After 13 months of treatment the patient is doing well with no tumor progression. CONCLUSIONS: This case appears to be one of the first documentations of beneficial effect and potential long-term benefit of TKIs in the treatment of fibrous histiocytoma. PMID- 19300421 TI - Limiting effect of diazepam on Lewis Lung carcinoma metastasis in anxious male mice. AB - AIM: It has been shown previously that chronic social defeat stress produces development of strong anxiety and increases intensity of experimental metastasis in the losers in comparison with the winners and control mice. The question was: is it possible to decrease the number of metastases in the losers by chronic or acute diazepam treatment. MATERIALS AND METHODS: Sensory contact model was used for generating male mice with repeated experience of social victories or defeats in daily agonistic interactions. Tumor cells of Lewis Lung Carcinoma (LLC) were injected into the tail vein of animals after 10 days of agonistic interactions. Then mice were treated acutely or chronically (7 days) with diazepam (1 mg/kg, i. p). Number of metastases in the lung was calculated in 16 days after tumor cell transplantation. RESULTS: Diazepam decreased the number of LLC metastases in anxious losers, whereas in the winners and control mice, without anxiety state, diazepam was ineffective. CONCLUSION: Well-known anxiolytic diazepam may decrease intensity of metastasis in anxious mice. PMID- 19300422 TI - Differential participation of angiotensin II type 1 and 2 receptors in the regulation of cardiac cell death triggered by angiotensin II. AB - BACKGROUND: The Angiotensin II (Ang II) type 1 (AT(1)R) and type 2 (AT(2)R) receptors are increased in the heart following myocardial infarction and dilated cardiomyopathy, yet their contribution at a cellular level to compensation and/or failure remains controversial. METHODS: We ectopically expressed AT(1)R and AT(2)R in cultured adult rat cardiomyocytes and cardiac fibroblasts to investigate Ang II-mediated cardiomyocyte hypertrophy and cardiac cell viability. RESULTS: In adult rat cardiomyocytes, Ang II did not induce hypertrophy via the AT(1)R, and no effect of Ang II on cell viability was observed following AT(1)R or AT(2)R expression. In adult rat cardiac fibroblasts, Ang II stimulated cell death by apoptosis via the AT(1)R (but not the AT(2)R), which required the presence of extracellular calcium, and induced a rapid dissipation of mitochondrial membrane potential, which was significant from 8 h. CONCLUSIONS: We conclude that Ang II/AT(1)R triggers apoptosis in adult rat cardiac fibroblasts, which is dependent on Ca2+ influx. PMID- 19300423 TI - Conjugated linoleic acid supplementation enhances antihypertensive effect of ramipril in Chinese patients with obesity-related hypertension. AB - BACKGROUND: Conjugated linoleic acid (CLA) refers to a group of positional and geometrical conjugated dienoic isomers of linoleic acid. Our aim was to investigate the effect of 8-week dietary CLA supplementation on blood pressure, concentrations of plasma adiponecin, leptin, and as well as angiotensin converting enzyme (ACE) activity in obese hypertensive subjects. METHODS: Eighty obese individuals with stage 1 uncontrolled essential hypertension were randomized in a double-blind, placebo-controlled trial. Participants were randomized to a daily dose of 4.5 g/day CLA (nine 0.5-g capsules; a 50:50 isomer blend of c 9,t 11 and t 10,c 12 CLA) with 37.5 mg/day ramipril (group 1) or placebo with 37.5 mg/day ramipril (group 2) for 8 weeks. Baseline and endpoint systolic BP, diastolic BP, and concentrations of plasma adiponecin, leptin, angiotensinogen, and ACE activity were measured. RESULTS: Treatment with CLA significantly enhanced the reduction effect of ramipril on systolic BP and diastolic BP (P < 0.05). It also increased plasma adiponectin concentration (P < 0.05) and decreased plasma concentrations of leptin and angiotensinogen (P < 0.05); however, significant change was not observed in ACE activity. CONCLUSIONS: An 8-week long supplementation of CLA enhanced the effect of ramipril on blood pressure reduction in treated obese hypertensive patients. The antihypertensive effect of CLA might be related to the changed secretion of hypertensive adipocytokines in plasma. PMID- 19300424 TI - Cluster-randomized controlled trial of oscillometric vs. manual sphygmomanometer for blood pressure management in primary care (CRAB). AB - BACKGROUND: Although mercury sphygmomanometers are seen as the gold standard instrument for blood pressure (BP) measurement, they are being withdrawn due to safety concerns. CRAB was a cluster-randomized controlled trial in 24 family practices in Tasmania, Australia, which aimed to determine the effect of an oscillometric device on BP management. METHODS: Cluster-randomized controlled trial. Intervention practices were supplied with automated monitors and control (usual care) practices used mercury or aneroid sphygmomanometers. They were subsequently audited by a research nurse. Usual care practice audit periods were matched to intervention practices. All analyses were intention-to-treat and adjusted for potential clustering. Differences in BP were analyzed using generalized estimating equations. All other outcomes were analyzed using multilevel mixed-effects Poisson regression. Post hoc analyses were performed to determine the mediators of changes in prescribing behavior. RESULTS: A total of 3,355 records were reviewed (828 visits had BP recordings). The percentage of BP recordings ending in "0" was significantly lower in intervention vs. usual care practices (systolic BP (SBP) 18% (107/587) vs. 71% (233/329), diastolic BP (DBP) 20% (119/584) vs. 70% (229/328), P < 0.001). The mean of SBP recordings in the intervention group was 7.5 mm Hg (95% confidence interval (CI) 5.2, 9.9 mm Hg, P < 0.001) higher than in the usual care group. Patients taking BP lowering drugs were more likely (incidence rate ratio (IRR) 1.3, 95% CI 1.1, 1.7, P = 0.01) to have a BP lowering drug prescribed if they were in the intervention compared to the usual care. CONCLUSIONS: Although digit preference was largely eliminated by oscillometric measurement, prescribing behavior was mediated by SBP. PMID- 19300425 TI - The impact of oral L-arginine supplementation on acute smoking-induced endothelial injury and arterial performance. AB - BACKGROUND: Smoking is associated with endothelial dysfunction and increased inflammatory status. The amino acid L-arginine, improves endothelial function in patients with cardiovascular risk factors. We investigated the effect of L arginine on vascular function and inflammatory process in healthy smokers at rest and after acute smoking. METHODS: We studied the effect of L-arginine and/or placebo in 12 healthy young smokers on three occasions (day 0, day 1, and day 3). The study was carried out on two separate arms, one with L-arginine (3 x 7 g/day) and one with placebo, according to a randomized, placebo-controlled, double blind, cross-over design. Measurements were carried out before, immediately after, and 20 min after cigarette smoking. Endothelial function was evaluated by flow-mediated dilation (FMD). Carotid-femoral pulse wave velocity (PWV) was measured as an index of aortic stiffness and augmentation index (AIx) and as a measure of arterial wave reflections. Serum soluble intercellular adhesion molecule-1 (sICAM-1) was measured. RESULTS: Compared to placebo, L-arginine improved FMD (P < 0.01 at day 1 and P < 0.05 at day 3). L-Arginine reduced PWV and AIx at both days 1 and 3 (P < 0.05 vs. baseline). L-Arginine blunted the acute smoking-induced increase of AIx at both day 1 (P < 0.05) and day 3 (P < 0.01), and prevented the smoking-induced elevation of PWV at day 3 (P < 0.05). Importantly, L-arginine reduced sICAM-1 at days 1 and 3 (P < 0.05 for both vs. baseline). CONCLUSIONS: Oral L-arginine improves endothelial function and vascular elastic properties of the arterial tree during the acute phase of smoking, an effect accompanied by reduced sICAM-1 levels in these subjects. PMID- 19300426 TI - Stress-related development of obesity and cortisol in women. AB - Chronic exposure to environmental stress may play a role in the development of obesity, through hyperactivation of the hypothalamic-pituitary-adrenocortical (HPA) axis. This study investigated the dynamics of weight gain and the activity of the HPA axis in women who developed weight gain after a stressful event. This is a case-control retrospective study. Two groups of age-matched premenopausal women were selected. One (n = 14) included women characterized by a rapid weight gain following a stressful event, defined as the "stress-related obesity " (SRO) group, and the other (n = 21) women with nonstress-related development of obesity, defined as the "nonstress-related obesity " (NSRO) group. Twenty-one healthy premenopausal women served as normal-weight controls. Baseline hormonal and metabolic parameters, and 24-h urinary free cortisol (UFC/24 h) excretion rate (as a measure of HPA-axis activity) were measured in all women. Anthropometry, diet, and physical activity were similar in both obese groups. Both obese groups showed similar metabolic and hormonal profiles, but the SRO group had UFC/24 h values (41.1 +/- 14.3 microg) significantly higher (P < 0.001) with respect to the NSRO (26.6 +/- 17.6 microg) or the normal-weight control groups (21.1 +/- 9.8 microg). Moreover, time (years) to achieve maximum Deltaweight gain (kg) and the Deltaweight gain/time ratio were significantly shorter (P < 0.001) and higher (P < 0.001) in the SRO group with respect to the NSRO group, respectively. In the SRO group, there was a tendency to a significant correlation between UFC/24 h and the Deltaweight gain/time ratio. These findings support the concept that SRO has distinct pathophysiological mechanisms, including hyperactivity of the HPA axis. PMID- 19300427 TI - Val1483Ile in FASN gene is linked to central obesity and insulin sensitivity in adult white men. AB - The Val1483Ile polymorphism in the human fatty acid synthase (FASN) gene is located within the interdomain region of the FASN close to the two dynamic active centers of the FASN enzyme and putatively affects FASN action. We aimed to evaluate the association of this polymorphism with obesity phenotypes, insulin sensitivity, and adipose tissue FASN activity in adult white subjects. The polymorphism was evaluated in association with metabolic variables in two independent studies: in a case-control study of 457 men (229 with normal glucose tolerance (NGT) and 228 with altered glucose tolerance (AGT)); and in 600 population-based NGT subjects (274 men and 326 women). Adipose tissue FASN activity was analyzed using the method of Nepokroeff. The Ile variant was associated with a lower waist-to-hip ratio (WHR) and a lower increase in weight over a 7-year period in NGT men. In a subset of 147 men, carriers of the Ile variant showed significantly increased insulin sensitivity. BMI (P < 0.001), WHR (P = 0.03), and Val1483Ile (P = 0.03), contributed independently to 37% of insulin sensitivity variance. In men from the population-based study, the Ile variant was associated with a lower BMI, WHR, fasting glucose, and systolic blood pressure compared with carriers of the Val variant. In agreement with these results, the adipose tissue FASN activity was significantly lower in subjects with the Ile variant (P = 0.01). In summary, adult white men with the Ile 1483 variant of the FASN gene seem protected from developing central obesity through decreased adipose tissue FASN activity. PMID- 19300428 TI - Differential effects of abdominal adipose tissue distribution on insulin sensitivity in black and white South African women. AB - Black South African women are more insulin resistant than BMI-matched white women. The objective of the study was to characterize the determinants of insulin sensitivity in black and white South African women matched for BMI. A total of 57 normal-weight (BMI 18-25 kg/m(2)) and obese (BMI > 30 kg/m(2)) black and white premenopausal South African women underwent the following measurements: body composition (dual-energy X-ray absorptiometry), body fat distribution (computerized tomography (CT)), insulin sensitivity (S(I), frequently sampled intravenous glucose tolerance test), dietary intake (food frequency questionnaire), physical activity (Global Physical Activity Questionnaire), and socioeconomic status (SES, demographic questionnaire). Black women were less insulin sensitive (4.4 +/- 0.8 vs. 9.5 +/- 0.8 and 3.0 +/- 0.8 vs. 6.0 +/- 0.8 x 10(-5)/min/(pmol/l), for normal-weight and obese women, respectively, P < 0.001), but had less visceral adipose tissue (VAT) (P = 0.051), more abdominal superficial subcutaneous adipose tissue (SAT) (P = 0.003), lower SES (P < 0.001), and higher dietary fat intake (P = 0.001) than white women matched for BMI. S(I) correlated with deep and superficial SAT in both black (R = -0.594, P = 0.002 and R = 0.495, P = 0.012) and white women (R = -0.554, P = 0.005 and R = -0.546, P = 0.004), but with VAT in white women only (R = -0.534, P = 0.005). In conclusion, body fat distribution is differentially associated with insulin sensitivity in black and white women. Therefore, the different abdominal fat depots may have varying metabolic consequences in women of different ethnic origins. PMID- 19300429 TI - A rare variant in the visfatin gene (NAMPT/PBEF1) is associated with protection from obesity. AB - Visfatin was recently reported as a novel adipokine encoded by the NAMPT (PBEF1) gene. This study was aimed at investigation of the possibility that single nucleotide polymorphisms (SNPs) in the visfatin gene are associated with either obesity or type 2 diabetes (T2D). A set of eight "tag-SNPs" were selected and ABI SNPlex assays designed for genotyping purposes. A total of 1,709 severely obese subjects were typed (896 class III obese adults and 813 children) together with 2,367 T2D individuals and 2,850 controls. For quantitative trait analysis, an additional 2,362 subjects were typed for rs10487818 from a general population sample. One rare SNP, rs10487818, located in intron 4 of NAMPT was associated with severe obesity, with a minor allele frequency of 1.6% in controls, 0.4% in the class III obese adults and, remarkably, 0% in the severely obese children. A highly significant association was observed for the presence or absence of the rare allele, i.e., (A,A) vs. (A,T + T,T) genotypes, in children (P = 6 x 10(-9)) and in adults (P = 8 x 10(-5)). No other significant (P < 0.05) association was observed with obesity or T2D for this or any other SNP. No association with BMI or waist-to-hip ratio was observed in a general population sample (n = 5,212). This is one of the first rare SNPs shown to be protective against a common polygenic disease and provides further evidence that rare alleles of strong effect can contribute to complex diseases such as severe obesity. PMID- 19300430 TI - Sexual orientation disparities in weight status in adolescence: findings from a prospective study. AB - A growing number of studies among adult women have documented disparities in overweight adversely affecting lesbian and bisexual women, but few studies have examined sexual orientation-related patterns in weight status among men or adolescents. We examined sexual orientation group trends in BMI (kg/m(2)), BMI Z scores, and overweight using 56,990 observations from 13,785 adolescent females and males in the Growing Up Today Study (GUTS), a large prospective cohort of US youth. Participants provided self-reported information from six waves of questionnaire data collection from 1998 to 2005. Gender-stratified linear regression models were used to estimate BMI and BMI Z-scores and modified Poisson regression models to estimate risk ratios for overweight, controlling for age and race/ethnicity, with heterosexuals as the referent group. Among females, we observed fairly consistently elevated BMI in all sexual orientation minority groups relative to heterosexual peers. In contrast, among males we documented a sexual-orientation-by-age interaction indicating steeper increases in BMI with age from early-to-late adolescence in heterosexuals relative to sexual orientation minorities. Additional prospective research is needed to understand the determinants of observed sexual orientation disparities and to inform appropriate preventive and treatment interventions. The long-term health consequences of overweight are well-documented and over time are likely to exact a high toll on populations with elevated rates. PMID- 19300431 TI - Gender, ethnicity, marital status, and body weight in the United States. AB - Married individuals tend to be heavier than those who are unmarried, particularly men, and individuals in different ethnic categories vary in their involvement in marriage and in their body weights. We examined gender and ethnic differences in relationships between marital status and body weight using cross-sectional data from the 1999-2002 National Health and Nutrition Examination Survey (NHANES) for 3,947 women and 4,019 men. The findings revealed that compared to married men in the same ethnic category, white divorced men, black never-married men, and all Hispanic men except for widows had lower odds of being overweight. Compared to married women in the same ethnic category, white women's weights did not significantly differ by marital status, black separated women had greater odds of being overweight, and Hispanic never-married women had lower odds of being overweight. Associations of marriage with body weight appear to be at least partly contingent upon gender and ethnicity, which may reflect larger societal patterns of involvement in marriage, commitment to family, and body-weight norms and expectations. PMID- 19300432 TI - Association of central obesity with the severity and audiometric configurations of age-related hearing impairment. AB - To investigate the effect of central obesity on the severity and characteristics of age-related hearing impairment (ARHI), we recruited 690 adult subjects with normal or symmetrical sensorineural hearing loss (SNHL). The effects of age, gender, morphometry, habits, systemic diseases, and environmental noise exposure on average pure tone hearing level at low frequencies (pure tone audiometry (PTA) low) and high frequencies (PTA-high) were analyzed. After adjusting for age, gender, systemic disease, and other variables, waist circumference (WC) showed a significant positive association with PTA-low and PTA-high. In females, PTA-low and PTA-high only showed significant positive association with age, but not with WC or other variables. However, PTA-high showed a positive association with borderline significance with WC in female subjects older than 55. In males, WC as well as age and noise exposure showed significant positive associations with both PTA-low and PTA-high, primarily in subjects younger than 55. When both WC and BMI were taken into account in a backward stepwise multivariate linear regression analysis, WC, but not BMI, showed a significant positive association with PTA-low and PTA-high in males younger than 55, and with PTA-high with borderline significance in females older than 55. However, the audiogram patterns were not significantly affected by central obesity in either age or gender. Our results suggest that WC was, even after adjustment for BMI, an independent risk factor of ARHI, particularly for low and high frequencies in males younger than 55 and for high frequencies in female subjects older than 55. PMID- 19300433 TI - The impact of obesity on health service utilization and costs in childhood. AB - Most studies of the economic costs of childhood obesity have focused upon hospitalization for comorbidities of obesity, whereas increased expenditures may also be the result of additional outpatient/emergency room visits or prescription drug expenditures. To quantify the magnitude of increased health-care utilization and expenditures among overweight and obese children, we performed descriptive, bivariate, and multivariable analyses on data from 6- to 19-year olds in the 2002 2005 Medical Expenditure Panel Survey (MEPS), a national probability survey of the noninstitutionalized civilian population in the United States. Compared with normal/underweight children, we found that children who were obese during both years of the MEPS had USD194 higher outpatient visit expenditures, USD114 higher prescription drug expenditures, and USD12 higher emergency room expenditures. Children who were overweight during both years, or overweight in one year and obese in the other had USD79 higher outpatient visit expenditures, USD64 higher prescription drug expenditures, and USD25 higher emergency room expenditures than normal/underweight children. Significantly, increased utilization was noted for outpatient visits, prescription drug use, and emergency room visits. Increased costs and utilization were concentrated among adolescents, though 6-11-year-old children who were obese in both years did have more outpatient visits and expenditures than other children. Extrapolated to the nation, elevated BMI in childhood was associated with USD14.1 billion in additional prescription drug, emergency room, and outpatient visit costs annually. Although further research is needed to identify effective interventions, the immediate economic consequences of childhood obesity are much greater than previously realized, and further reinforce efforts to prevent this major comorbidity are needed. PMID- 19300434 TI - How physician obesity specialists use drugs to treat obesity. AB - Specialist physicians may have prescribing habits that are different from nonspecialist physicians. Little is known about the prescribing habits of physicians specializing in the treatment of obesity. An anonymous survey was given to the physician members of the American Society of Bariatric Physicians (ASBP). There was a 35% response rate (266 physicians) to the questionnaire that was represented nationally. Almost all prescribed medications and all of them recommended phentermine. The average maximal dose of phentermine was above that approved in the package insert, and these physicians disagreed with the National Institutes of Health (NIH) Obesity Treatment Guidelines. Phendimetrazine, metformin, and phentermine plus L-5-hydroxytryptophan (5-HTP) with carbidopa were all used more frequently than either orlistat or sibutramine. The combination of sibutramine and orlistat as well as 5-HTP/carbidopa were prescribed by 14 and 20%, respectively. As 5-HTP-carbidopa was a combination not previously reported for the treatment of obesity, a retrospective chart review was performed in a single obesity practice, which may not be representative. Twenty-two subjects had a 16% weight loss with phentermine over 6 months and an additional 1% weight loss with the addition of 5-HTP/carbidopa for an additional 6 months. One subject who started on 5-HTP/carbidopa alone lost 24.4% of initial body weight over 6 months. This questionnaire revealed that 20% of the obesity specialists responding to the survey used phentermine plus of 5-HTP/carbidopa, an unreported combination. A controlled, randomized, clinical trial to evaluate the safety and efficacy of this combination in treating obesity should be considered. PMID- 19300435 TI - The obesity epidemic and future emergency responders. AB - Emergency responders should be fit to safely perform strenuous duties. In particular, young recruits are expected to be at or near peak career fitness. We studied the prevalence and health associations of excess weight among 370 consecutive emergency responder candidates for fire and ambulance services in Massachusetts. The mean age and BMI of the recruits were 26.3 (3.8) years and 28.5 (4.9) kg/m(2), respectively. Seventy-seven percent had BMI >or=25 kg/m(2), and 33% were obese (BMI >or=30 kg/m(2)). After multivariate adjustment, both higher BMI categories and unit increases in BMI were significantly associated with higher blood pressures, worse metabolic profiles, and lower exercise tolerance. Excess weight is highly prevalent and associated with elevated cardiovascular risk among future emergency responders. These findings in a population expected to perform demanding duties supporting public safety merit prompt public health intervention. PMID- 19300436 TI - The Japanese Society of Hypertension Guidelines for the Management of Hypertension (JSH 2009). PMID- 19300437 TI - The structural basis for peptide selection by the transport receptor OppA. AB - Oligopeptide-binding protein A (OppA) from Lactococcus lactis binds peptides of an exceptionally wide range of lengths (4-35 residues), with no apparent sequence preference. Here, we present the crystal structures of OppA in the open- and closed-liganded conformations. The structures directly explain the protein's phenomenal promiscuity. A huge cavity allows binding of very long peptides, and a lack of constraints for the position of the N and C termini of the ligand is compatible with binding of peptides with varying lengths. Unexpectedly, the peptide's amino-acid composition (but not the exact sequence) appears to have a function in selection, with a preference for proline-rich peptides containing at least one isoleucine. These properties can be related to the physiology of the organism: L. lactis is auxotrophic for branched chain amino acids and favours proline-rich caseins as a source of amino acids. We propose a new mechanism for peptide selection based on amino-acid composition rather than sequence. PMID- 19300438 TI - Mimicking Ndc80 phosphorylation triggers spindle assembly checkpoint signalling. AB - The protein kinase Mps1 is, among others, essential for the spindle assembly checkpoint (SAC). We found that Saccharomyces cerevisiae Mps1 interacts physically with the N-terminal domain of Ndc80 (Ndc80(1-257)), a constituent of the Ndc80 kinetochore complex. Furthermore, Mps1 effectively phosphorylates Ndc80(1-257) in vitro and facilitates Ndc80 phosphorylation in vivo. Mutating 14 of the phosphorylation sites to alanine results in compromised checkpoint signalling upon nocodazole treatment of mutants. Mutating the identical sites to aspartate (to simulate constitutive phosphorylation) causes a metaphase arrest with wild-type-like bipolar kinetochore-microtubule attachment. This arrest is due to a constitutively active SAC and consequently the inviable aspartate mutant can be rescued by disrupting SAC signalling. Therefore, we conclude that a putative Mps1-dependent phosphorylation of Ndc80 is important for SAC activation at kinetochores. PMID- 19300439 TI - Acid sphingomyelinase activity triggers microparticle release from glial cells. AB - We have earlier shown that microglia, the immune cells of the CNS, release microparticles from cell plasma membrane after ATP stimulation. These vesicles contain and release IL-1beta, a crucial cytokine in CNS inflammatory events. In this study, we show that microparticles are also released by astrocytes and we get insights into the mechanism of their shedding. We show that, on activation of the ATP receptor P2X7, microparticle shedding is associated with rapid activation of acid sphingomyelinase, which moves to plasma membrane outer leaflet. ATP induced shedding and IL-1beta release are markedly reduced by the inhibition of acid sphingomyelinase, and completely blocked in glial cultures from acid sphingomyelinase knockout mice. We also show that p38 MAPK cascade is relevant for the whole process, as specific kinase inhibitors strongly reduce acid sphingomyelinase activation, microparticle shedding and IL-1beta release. Our results represent the first demonstration that activation of acid sphingomyelinase is necessary and sufficient for microparticle release from glial cells and define key molecular effectors of microparticle formation and IL-1beta release, thus, opening new strategies for the treatment of neuroinflammatory diseases. PMID- 19300440 TI - Redox-switch modulation of human SSADH by dynamic catalytic loop. AB - Succinic semialdehyde dehydrogenase (SSADH) is involved in the final degradation step of the inhibitory neurotransmitter gamma-aminobutyric acid by converting succinic semialdehyde to succinic acid in the mitochondrial matrix. SSADH deficiency, a rare autosomal recessive disease, exhibits variable clinical phenotypes, including psychomotor retardation, language delay, behaviour disturbance and convulsions. Here, we present crystal structures of both the oxidized and reduced forms of human SSADH. Interestingly, the structures show that the catalytic loop of the enzyme undergoes large structural changes depending on the redox status of the environment, which is mediated by a reversible disulphide bond formation between a catalytic Cys340 and an adjacent Cys342 residues located on the loop. Subsequent in vivo and in vitro studies reveal that the 'dynamic catalytic loop' confers a response to reactive oxygen species and changes in redox status, indicating that the redox-switch modulation could be a physiological control mechanism of human SSADH. Structural basis for the substrate specificity of the enzyme and the impact of known missense point mutations associated with the disease pathogenesis are presented as well. PMID- 19300441 TI - Laboratory procedures to generate viral metagenomes. AB - This collection of laboratory protocols describes the steps to collect viruses from various samples with the specific aim of generating viral metagenome sequence libraries (viromes). Viral metagenomics, the study of uncultured viral nucleic acid sequences from different biomes, relies on several concentration, purification, extraction, sequencing and heuristic bioinformatic methods. No single technique can provide an all-inclusive approach, and therefore the protocols presented here will be discussed in terms of hypothetical projects. However, care must be taken to individualize each step depending on the source and type of viral-particles. This protocol is a description of the processes we have successfully used to: (i) concentrate viral particles from various types of samples, (ii) eliminate contaminating cells and free nucleic acids and (iii) extract, amplify and purify viral nucleic acids. Overall, a sample can be processed to isolate viral nucleic acids suitable for high-throughput sequencing in approximately 1 week. PMID- 19300442 TI - Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics. AB - Accurate quantification of protein expression in biological systems is an increasingly important part of proteomics research. Incorporation of differential stable isotopes in samples for relative protein quantification has been widely used. Stable isotope incorporation at the peptide level using dimethyl labeling is a reliable, cost-effective and undemanding procedure that can be easily automated and applied in high-throughput proteomics experiments. Although alternative multiplex quantitative proteomics approaches introduce isotope labels at the organism level ('stable isotope labeling by amino acids in cell culture' (SILAC)) or enable the simultaneous analysis of eight samples (isobaric tagging for relative and absolute quantification (iTRAQ)), stable isotope dimethyl labeling is advantageous in that it uses inexpensive reagents and is applicable to virtually any sample. We describe in-solution, online and on-column protocols for stable isotope dimethyl labeling of sample amounts ranging from sub micrograms to milligrams. The labeling steps take approximately 60-90 min, whereas the full protocol including digestion and (two-dimensional) liquid chromatography-mass spectrometry takes approximately 1.5-3 days to complete. PMID- 19300443 TI - Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors. AB - Over the past decade, lentiviral vectors have emerged as powerful tools for transgene delivery. The use of lentiviral vectors has become commonplace and applications in the fields of neuroscience, hematology, developmental biology, stem cell biology and transgenesis are rapidly emerging. Also, lentiviral vectors are at present being explored in the context of human clinical trials. Here we describe improved protocols to generate highly concentrated lentiviral vector pseudotypes involving different envelope glycoproteins. In this protocol, vector stocks are prepared by transient transfection using standard cell culture media or serum-free media. Such stocks are then concentrated by ultracentrifugation and/or ion exchange chromatography, or by precipitation using polyethylene glycol 6000, resulting in vector titers of up to 10(10) transducing units per milliliter and above. We also provide reliable real-time PCR protocols to titrate lentiviral vectors based on proviral DNA copies present in genomic DNA extracted from transduced cells or on vector RNA. These production/concentration methods result in high-titer vector preparations that show reduced toxicity compared with lentiviral vectors produced using standard protocols involving ultracentrifugation-based methods. The vector production and titration protocol described here can be completed within 8 d. PMID- 19300444 TI - Complex pathogenesis of Hirschsprung's disease in a patient with hydrocephalus, vesico-ureteral reflux and a balanced translocation t(3;17)(p12;q11). AB - Hirschsprung's disease (HSCR), a congenital complex disorder of intestinal innervation, is often associated with other inherited syndromes. Identifying genes involved in syndromic HSCR cases will not only help understanding the specific underlying diseases, but it will also give an insight into the development of the most frequent isolated HSCR. The association between hydrocephalus and HSCR is not surprising as a large number of patients have been reported to show the same clinical association, most of them showing mutations in the L1CAM gene, encoding a neural adhesion molecule often involved in isolated X linked hydrocephalus. L1 defects are believed to be necessary but not sufficient for the occurrence of the intestinal phenotype in syndromic cases. In this paper, we have carried out the molecular characterization of a patient affected with Hirschsprung's disease and X-linked hydrocephalus, with a de novo reciprocal balanced translocation t(3;17)(p12;q21). In particular, we have taken advantage of this chromosomal defect to gain access to the predisposing background possibly leading to Hirschsprung's disease. Detailed analysis of the RET and L1CAM genes, and molecular characterization of MYO18A and TIAF1, the genes involved in the balanced translocation, allowed us to identify, besides the L1 mutation c.2265delC, different additional factors related to RET-dependent and independent pathways which may have contributed to the genesis of enteric phenotype in the present patient. PMID- 19300446 TI - The anatomy of memory: an interactive overview of the parahippocampal-hippocampal network. AB - Converging evidence suggests that each parahippocampal and hippocampal subregion contributes uniquely to the encoding, consolidation and retrieval of declarative memories, but their precise roles remain elusive. Current functional thinking does not fully incorporate the intricately connected networks that link these subregions, owing to their organizational complexity; however, such detailed anatomical knowledge is of pivotal importance for comprehending the unique functional contribution of each subregion. We have therefore developed an interactive diagram with the aim to display all of the currently known anatomical connections of the rat parahippocampal-hippocampal network. In this Review, we integrate the existing anatomical knowledge into a concise description of this network and discuss the functional implications of some relatively underexposed connections. PMID- 19300445 TI - Transient cell-cell interactions in neural circuit formation. AB - The wiring of the nervous system requires a complex orchestration of developmental events. Emerging evidence suggests that transient cell-cell interactions often serve as positional cues for axon guidance and synaptogenesis during the assembly of neural circuits. In contrast to the relatively stable cellular interactions between synaptic partners in mature circuits, these transient interactions involve cells that are not destined to be pre- or postsynaptic cells. Here we review the roles of these transient cell-cell interactions in a variety of developmental contexts and describe the mechanisms through which they organize neural connections. PMID- 19300447 TI - Effects of eplerenone, a selective mineralocorticoid receptor antagonist, on clinical and experimental salt-sensitive hypertension. AB - Mineralocorticoid receptors (MRs) are expressed in non-epithelial tissues, such as blood vessels, the heart and adipose tissue. The combined effects of aldosterone and insulin link the metabolic syndrome with hypertension and salt sensitivity. Eplerenone is the newly developed inhibitor of MRs that has significantly fewer adverse effects than similar doses of spironolactone. Eplerenone has been reported to have anti-hypertensive and protective effects on cardiovascular and renal injury in salt-sensitive hypertensive animal models, such as the Dahl salt-sensitive (DS) hypertensive rat and leptin receptor deficient spontaneously hypertensive rat (SHR/cp). Eplerenone also increases nitric oxide bioavailability and improves impaired endothelial function by decreasing oxidative stress. Clinical studies support the concept that eplerenone is effective for the treatment of salt-sensitive hypertension as well as idiopathic hyperaldosteronism and does not have adverse anti-androgenic adverse effects. In Japan, eplerenone has been used clinically since 2007 for the treatment of hypertension, with its price being marginally lower than all types of angiotensin II receptor antagonists. This will inevitably result in an increasing number of hypertensive patients and those with primary aldosteronism being treated with this agent in the near future. PMID- 19300448 TI - Acute and chronic metabolic acidosis interferes with aquaporin-2 translocation in the rat kidney collecting ducts. AB - Renal aquaporin-2 (AQP2) expression plays a key role in urine concentration. However, it is not known whether metabolic acidosis affects urine-concentrating ability through AQP2 expression in the kidney and urine. We examined urinary excretion and renal expression of AQP2 in control and acidosis rats, using RT competitive PCR, immunoblot and immunocytochemistry. Urinary excretion of AQP2 is decreased by 92% even with the increase in AQP2 mRNA and protein expressions in the collecting ducts by metabolic acidosis in rats. Urine osmolality in control rats was 1670+/-198 mOsm per kg H(2)O, and immunocytochemistry revealed the presence of AQP2 in the apical plasma membrane of the principal cells in the collecting ducts. Urine osmolality in acidosis rats was lower than that in control (1397+/-243 mOsm per kg H(2)O), and immunocytochemistry showed the diffuse presence of AQP2 in the cytoplasm of the principal cells. Differential centrifugation-coupled immunoblot showed a significant decrease in the ratio of AQP2 in plasma membrane-enriched fraction to that in intracellular vesicle enriched fraction by metabolic acidosis. In summary, AQP2 translocation is largely decreased by metabolic acidosis even with increased expression in the collecting ducts. A disorder of AQP2 translocation in the collecting ducts with acidosis may be responsible for the diuresis in patients with chronic renal failure. PMID- 19300449 TI - Loss of nocturnal decline of blood pressure in non-diabetic patients with nephrotic syndrome in the early and middle stages of chronic kidney disease. AB - In non-diabetic patients with nephrotic syndrome (NS) at early stages of chronic kidney disease, it remains unclear whether the degree of proteinuria affects the nocturnal blood pressure (BP) dip. We evaluated the relationship among circadian BP rhythm, proteinuria and hypoalbuminemia in these patients. We also evaluated the autonomic nervous activity. Twenty-four-hour BP was measured in NS patients (8 men and 13 women; mean age, 58.5+/-14.8 years) and age- and sex-matched normal subjects (11 men and 13 women; mean age, 54.3+/-18.2 years) as controls. Serum albumin and urinary protein concentrations were measured. Power spectral analysis of the heart rate was performed, and the high frequency (HF) and low frequency (LF) components were calculated as indices of sympathovagal balance. There were no differences in waking BP between the NS and the control groups (131+/-13/78+/ 9 vs. 130+/-17/76+/-7 mm Hg; P>0.05). However, sleeping BP was significantly higher in the NS group than in the control group (127+/-18/75+/-9 vs. 115+/ 14/66+/-7 mm Hg; P<0.05). Sleeping/waking BP ratios were higher in the NS group than in the control group (P<0.01). In the NS group, these ratios correlated significantly with serum albumin level (r=-0.54, P=0.011 for systolic BP; r= 0.48, P=0.030 for diastolic BP) and urinary protein excretion (r=0.47, P=0.027 for systolic BP; r=0.60, P=0.003 for diastolic BP). Both HF components and LF/HF ratios were not significantly different between the two groups. In non-diabetic NS patients, loss of nocturnal BP reduction correlates with proteinuria and hypoalbuminemia but not with circadian autonomic nervous rhythm. PMID- 19300450 TI - Effects of metoprolol on epinephrine-induced takotsubo-like left ventricular dysfunction in non-human primates. AB - Takotsubo cardiomyopathy, alternatively known as stress cardiomyopathy, is an increasingly recognized clinical syndrome characterized by acute reversible apical ventricular dysfunction. To elucidate the mechanism, we tried to make a new model of takotsubo-like cardiomyopathy in non-human primates. Echocardiography revealed that repeated intravenous infusion of epinephrine overdose in cynomolgus monkeys induced takotsubo-like cardiomyopathy, which is characterized by progressive left ventricle and depressed systolic function with severe hypokinesis in apical regions and hyperkinesis in the basal region. Although this cardiac dysfunction almost normalized after a month even without any treatment, metoprolol, a beta-blocker, improved the decreased ejection fraction earlier than in the control. Luxol fast blue staining, which is useful for estimating myocytolysis, showed that increased myocytolysis was observed in the apical ventricle of the epinephrine-infused heart. Metoprolol diminished epinephrine-induced cardiomyocytolysis. To explain the mechanism of takotsubo myopathy and the effect of metoprolol, gene expressions in apical or basal ventricle were compared. Heart failure-related genes, such as brain natriuretic peptide, connective tissue growth factor and osteopontin; calcium signaling related genes, such as ryanodine receptor 2, sarcoendoplasmic reticulum Ca(2+) ATPase 2A2 and adenylate cyclase 7; renin-angiotensin system-related genes, such as angiotensinogen, angiotensin II receptor, type 1 and type 2; and mitochondria related genes, such as peroxisome proliferator-activated receptor-gamma co activator-1alpha, cytochrome c and transcription factor A mitochondrial, were significantly changed at the apical ventricle rather than at the basal ventricle. The changes of some genes improved with metoprolol treatment. These results indicate that this model is valuable in understanding the pathogenesis of takotsubo cardiomyopathy and the effectivity of beta-blockers. PMID- 19300451 TI - The expression of matrix metalloproteinase-13 is increased in vessels with blood brain barrier impairment in a stroke-prone hypertensive model. AB - We previously reported that the blood-brain barrier (BBB) function was deteriorated in vessels located in the hippocampus, but not the cerebral cortex, in 3-month-old stroke-prone spontaneously hypertensive rats (SHRSP). Recently published data suggest that matrix metalloproteinase (MMP)-2 and MMP-9 play a critical role in the BBB disruption in stroke or cerebral ischemia. In this study, we examined gene and protein expressions of MMPs in the BBB-damaged hippocampal vessels of 3-month-old SHRSP, in the cerebral cortical vessels without BBB damage of SHRSP, and in the hippocampal and cerebral cortical ones without BBB damage of 3-month-old Wistar Kyoto (WKY) rats. The expressions of MMPs were examined by real-time quantitative reverse transcriptase-PCR (RT-PCR), western blotting and immunohistochemical techniques. The gene and protein expressions of MMP-13 were significantly increased in the hippocampal samples of SHRSP compared with samples without BBB damage, such as cerebral cortical samples of SHRSP or hippocampal samples of WKY. Immunostaining of MMP-13 was seen in the cytoplasm of ED-1-positive perivascular cells in both rats and was colocalized with those of type IV collagen or osteopontin. The type IV collagen was also localized in the basement membrane. These findings indicate that the expression of MMP-13 is increased in BBB-damaged hippocampal vessels in hypertensive SHRSP compared with vessels without BBB impairment in normotensive WKY rats and may be involved in vascular remodeling. PMID- 19300452 TI - PUMA- and Bax-induced autophagy contributes to apoptosis. AB - The p53-inducible BH3-only protein PUMA is a key mediator of p53-dependent apoptosis, and PUMA has been shown to function by activating Bax and mitochondrial outer membrane permeabilization. In this study, we describe an ability of PUMA to induce autophagy that leads to the selective removal of mitochondria. This function of PUMA depends on Bax/Bak and can be reproduced by overexpression of Bax. The induction of autophagy coincides with cytochrome c release, and taken together the results suggest that PUMA functions through Bax to induce mitochondrial autophagy in response to mitochondrial perturbations. Surprisingly, inhibition of PUMA or Bax-induced autophagy dampens the apoptotic response, suggesting that under some circumstances the selective targeting of mitochondria for autophagy can enhance apoptosis. PMID- 19300453 TI - The alpha/beta carboxy-terminal domains of p63 are required for skin and limb development. New insights from the Brdm2 mouse which is not a complete p63 knockout but expresses p63 gamma-like proteins. AB - p63, an ancestral transcription factor of the p53 family, has three C-terminal isoforms whose relative in vivo functions are elusive. The p63 gene is essential for skin and limb development, as vividly shown by two independent global knockout mouse models. Both strains, although constructed differently, have identical and severe phenotypes, characterized by absent epidermis and hindlimbs and only rudimentary forelimbs at birth. Here we show that mice from one model, Brdm2, express normal levels of truncated p63 proteins that contain the DNA binding and oligomerization domain but lack the long carboxy-terminal SAM (sterile alpha-motif) and post-SAM domains that are specific for the alpha and beta isoforms. As such, transcriptionally active p63 proteins from Brdm2 mice resemble the naturally occurring p63gamma isoforms, which of all the p63 isoforms most closely resemble p53. Thus, Brdm2 mice are p63alpha/beta isoform-specific knockout mice, gaining unexpected new importance. Our studies identify that p63alpha/beta but not p63gamma are absolutely required for proper skin and limb development. PMID- 19300454 TI - Genetic deletion of faim reveals its role in modulating c-FLIP expression during CD95-mediated apoptosis of lymphocytes and hepatocytes. AB - Fas-apoptosis inhibitory molecule (FAIM) is inducibly expressed in B lymphocytes and had been shown to antagonize Fas-mediated killing of B-cell lines in vitro. However, its mechanism and role in vivo are unknown. We have generated faim(-/-) mice and found these mutants to be viable. In contrast to fas(-/-) mice, faim(-/ ) mice have normal B- and T-cell populations. However, faim(-/-) B cells and thymocytes show increased sensitivity to Fas-triggered apoptosis in vitro, and faim(-/-) mice suffer greater mortality and exhibit exacerbated liver damage in response to Fas (CD95) engagement in vivo. The lack of FAIM results in greater activation of caspase-8 and -3 in Fas-stimulated thymocytes. Detailed biochemical analyses further reveal the decreased expression of c-FLIP(L) and c-FLIP(R) in faim(-/-) thymocytes and increased association of caspase-8 with Fas in Fas activated mutant cells. Decreased levels of c-FLIP(L) and c-FLIP(R) are also evident in faim(-/-) liver. Thus, FAIM plays a novel role in modulating Fas mediated apoptosis and acts through influencing the expression of c-FLIP and regulating the physical binding of caspase-8 to Fas. PMID- 19300455 TI - The E3 ubiquitin ligase specificity subunit ASB2beta is a novel regulator of muscle differentiation that targets filamin B to proteasomal degradation. AB - Ubiquitin-mediated protein degradation is the main mechanism for controlled proteolysis, which is crucial for muscle development and maintenance. The ankyrin repeat-containing protein with a suppressor of cytokine signaling box 2 gene (ASB2) encodes the specificity subunit of an E3 ubiquitin ligase complex involved in differentiation of hematopoietic cells. Here, we provide the first evidence that a novel ASB2 isoform, ASB2beta, is important for muscle differentiation. ASB2beta is expressed in muscle cells during embryogenesis and in adult tissues. ASB2beta is part of an active E3 ubiquitin ligase complex and targets the actin binding protein filamin B (FLNb) for proteasomal degradation. Thus, ASB2beta regulates FLNb functions by controlling its degradation. Knockdown of endogenous ASB2beta by shRNAs during induced differentiation of C2C12 cells delayed FLNb degradation as well as myoblast fusion and expression of muscle contractile proteins. Finally, knockdown of FLNb in ASB2beta knockdown cells restores myogenic differentiation. Altogether, our results suggest that ASB2beta is involved in muscle differentiation through the targeting of FLNb to destruction by the proteasome. PMID- 19300456 TI - Complex II inhibition by 3-NP causes mitochondrial fragmentation and neuronal cell death via an NMDA- and ROS-dependent pathway. AB - Mitochondrial respiratory complex II inhibition plays a central role in Huntington's disease (HD). Remarkably, 3-NP, a complex II inhibitor, recapitulates HD-like symptoms. Furthermore, decreases in mitochondrial fusion or increases in mitochondrial fission have been implicated in neurodegenerative diseases. However, the relationship between mitochondrial energy defects and mitochondrial dynamics has never been explored in detail. In addition, the mechanism of neuronal cell death by complex II inhibition remains unclear. Here, we tested the temporal and spatial relationship between energy decline, impairment of mitochondrial dynamics, and neuronal cell death in response to 3-NP using quantitative fluorescence time-lapse microscopy and cortical neurons. 3-NP caused an immediate drop in ATP. This event corresponded with a mild rise in reactive oxygen species (ROS), but mitochondrial morphology remained unaltered. Unexpectedly, several hours after this initial phase, a second dramatic rise in ROS occurred, associated with profound mitochondrial fission characterized by the conversion of filamentous to punctate mitochondria and neuronal cell death. Glutamate receptor antagonist AP5 abolishes the second peak in ROS, mitochondrial fission, and cell death. Thus, secondary excitotoxicity, mediated by glutamate receptor activation of the NMDA subtype, and consequent oxidative and nitrosative stress cause mitochondrial fission, rather than energy deficits per se. These results improve our understanding of the cellular mechanisms underlying HD pathogenesis. PMID- 19300457 TI - Acquisition of inverted GSTM exons by an intron of primate GSTM5 gene. AB - The human GSTM gene family is composed of five gene members, GSTM1-5, and plays an important role in detoxification. In this study, the human GSTM5 gene was found to have a long inverted repeat (LIR) in intron 5. The LIR is able to form a stem-loop structure with a 31-bp stem and a 9-nt loop. The intronic LIR was also identified in other primates but not in non-primates. The human and chimpanzee LIRs had undergone compensating mutations that make the stem loop more stable, suggesting a functional role for the LIR. Sequence homology showed that the LIR was actually a part of inverted exons acquired by the intron. Results of phylogenetic analysis indicate that the inverted exons were derived from exon 5 of GSTM4 and exon 5 of GSTM1. The intronic LIR and inverted GSTM exons can probably introduce complexity in the expression of GSTM gene family. PMID- 19300458 TI - Physicians' opinion for 'new' genetic testing in Japan. AB - Recent advances in studies on human genetics have led to the use of genetic information in various applications. We conducted a survey to know the opinions of healthcare providers in Japan on new genetic testing services, such as direct to-consumer (DTC) genetic testing. A total of 1124 general practitioners and 294 clinical geneticists replied to our questionnaire. Thirty-eight percent of the general practitioners and 68.4% of the clinical geneticists were aware of DTC genetic testing. Some physicians had gained information on this service through their patients or commercial activities of companies providing such services. General practitioners expected that DTC genetic testing would be convenient, promote preventive medicine, provide personalized services and would enable to maintain confidentiality of information. Clinical geneticists showed greater concern with regard to the reliability of the results, provision of information/counseling and the understanding of results. Awareness of DTC genetic testing enhances general practitioners' positive opinions of it. Although the market for DTC genetic testing in Japan may still be limited, it is possible that general practitioners will play a role in the provision of DTC genetic testing services in the future. On the basis of their knowledge and experience, clinical geneticists should provide information to both healthcare providers and to the public. PMID- 19300459 TI - The microeconomics of personalized medicine: today's challenge and tomorrow's promise. AB - 'Personalized medicine' promises to increase the quality of clinical care and, in some cases, decrease health-care costs. Despite this, only a handful of diagnostic tests have made it to market, with mixed success. Historically, the challenges in this field were scientific. However, as discussed in this article, with the maturation of the '-omics' sciences, it now seems that the major barriers are increasingly related to economics. Overcoming the poor microeconomic alignment of incentives among key stakeholders is therefore crucial to catalysing the further development and adoption of personalized medicine, and we propose several actions that could help achieve this goal. PMID- 19300461 TI - 'The Microhand': a new concept of micro-forceps for ocular robotic surgery. AB - PURPOSE: To test the feasibility of retinal manipulations using a new micromanipulator (Microhand) for ocular robotic microsurgery. METHODS: Pneumatically actuated four-finger microhands were developed at UCLA with micro electromechanical systems (MEMS) technology to mimic a human hand for small object manipulation. Microhands with four 4 mm finger lengths were used for this study to lift caliper weights and fresh retinal tissue of porcine cadaver eyes to find the maximum force at a given pressure and feasibility of the microhands for retinal manipulation in real surgery. RESULTS: A full closure of the microhand used for caliper weight lifting was achieved under 65 psi (448 kPa) of air pressure. The four-fingered microhand was able to develop about 20 mN of total lifting force and 5 mN per finger at 80 psi (551 kPa), and was strong enough to displace and lift the retina of pig eyes. CONCLUSIONS: The microhand is able to apply calibrated forces to ocular tissues and is suitable for ocular microsurgical procedures. This new tool would be useful in the development of robotic microsurgery. PMID- 19300460 TI - Chemical modulators of sphingosine-1-phosphate receptors as barrier-oriented therapeutic molecules. AB - Biological barriers regulate the passage of cells, pathogens, fluids, nutrients, ions and signalling molecules between anatomical compartments during homeostasis and disease. Yet strategies that allow for reversible therapeutic modulation of these barriers are still in their infancy. The enhancement or protection of natural barriers is desirable in conditions such as acute respiratory distress syndrome or ischaemia-reperfusion injuries, whereas a temporary disruption could facilitate the penetration of drugs across such barriers. This Review discusses the role of sphingosine-1-phosphate receptors in the regulation and protection of biological barriers, and the potential of therapeutic strategies that target this receptor family. PMID- 19300462 TI - In vivo lamellar keratoplasty using platelet-rich plasma as a bioadhesive. AB - PURPOSE: The use of standard sutures has been replaced by platelet-rich plasma (PRP), a bioadhesive agent, in several surgical procedures. This prompted us to test PRP efficacy in experimental lamellar keratoplasty. METHODS: After lamellar anterior keratoplasty, PRP with a mean concentration of 807,564 platelets/mm(3) was used to attach the corneal flap to the stromal surface in 12 New Zealand white rabbits. 10-0 nylon sutures were used in one control group of 12 animals and no suture was used in a second control group of six rabbits. Animals were killed at days 2, 7, 30, and 90 for histological and smooth muscle actin (SMA) immunohistochemical analysis. RESULTS: The PRP group showed a tight corneal graft from the first postoperative hours until they were killed. A transparent cornea was seen at 30 days and remained clear until the end of the protocol. Histological specimens showed no signs of ocular inflammation in any animal within the PRP group. Electron microscopy showed normal morphological features on the flap and stromal bed, and a clear interface zone without cells or debris. The number of stromal myofibroblasts was lower than that seen in the suture group at 3 months postsurgery. The sutured group showed an attached cornea with signs of inflammation around the knots. All flaps without PRP or sutures were completely detached in the immediate postoperative period. CONCLUSIONS: PRP was useful for attaching the corneal flap and it was well tolerated by the rabbit corneal tissue. Corneal healing was satisfactory. Further studies on PRP adhesiveness in grafts with donor corneas should be performed before considering its use in patients. PMID- 19300463 TI - The value of an immune response to Mycobacterium tuberculosis in patients with chronic posterior uveitis revisited: utility of the new IGRAs. AB - PURPOSE: To explore the utility of a specific immune response to Mycobacterium tuberculosis in a population of immunosuppressed idiopathic chronic posterior uveitis patients, by means of a tuberculosis-specific interferon-gamma release assay. DESIGN: Prospective, interventional case series. METHODS: A total of 31 referred patients with severe idiopathic chronic uveitis or panuveitis and 52 controls were screened for a specific immune response to tuberculosis. After ruling-out specific uveitis entities, presumed tuberculosis-related uveitis was initially considered when ophthalmologic findings were consistent with tubercular uveitis, and a specific immune response to M. tuberculosis confirmed by QuantiFERON, despite inability to detect M. tuberculosis. Clinical responses to antitubercular treatment were recorded. RESULTS: The prevalence of an immune response to M. tuberculosis was 15.38% in controls and 32.25% in uveitis patients (OR=2.619, P=0.07). Two patients were QuantiFERON indeterminate (6.4%). After excluding seven specific uveitis entities (OR=3.66, P=0.03), eight QuantiFERON positive and one QuanTIFERON-negative uveitis patients were initially treated for presumed tuberculosis-related uveitis. All but one had no evidence of active systemic involvement. None had been previously diagnosed with tuberculosis, but unsuccessfully treated with immunosuppressors. After a 9-month tuberculostatic treatment, seven QuantiFERON -positive and one QuantiFERON-negative patients exhibited decreased intraocular inflammation, visual acuity improvement, and no relapses. Estimated QuantiFERON sensitivity and specificity were 82 and 100%, respectively, with a PPV=100% and an NPV=86%. CONCLUSIONS: QuantiFERON was useful for antituberculous treatment decision-making in chronic posterior uveitis immunosuppressed patients from areas with an intermediate-high prevalence of tuberculosis. PMID- 19300464 TI - Corticosteroid-induced intraocular pressure elevation in keratoconus is common following uncomplicated penetrating keratoplasty. AB - AIMS: To determine the incidence of postkeratoplasty intraocular pressure (IOP) elevation in the eyes of subjects with keratoconus and establish the relationship between IOP and corticosteroid administrations in this population. METHODS: Following strict inclusion/exclusion criteria, a retrospective analysis was performed on a consecutive series of penetrating keratoplasties performed for keratoconus observing a standardised surgical and postoperative regimen in Auckland, New Zealand. Patient demographics, ocular, medical, and family history, and pre- and postoperative data were recorded until 12 months postkeratoplasty. RESULTS: In all, 57 eyes of 48 patients were included-31% New Zealand Europeans, 42% Pacific people, 15% Maori, and 12% other. Eighteen eyes (32%) of 17 patients (35%) exhibited elevated IOP and 12 (21%) eyes exhibited moderate-to-severe elevation of IOP. IOP elevation occurred 3-6 months postkeratoplasty in 78% of eyes. Elevated IOP was significantly less common in Maori and Pacific peoples (P=0.02). All eyes except one required reduction/cessation of corticosteroids to normalise IOP. CONCLUSIONS: The incidence of presumed steroid-related postkeratoplasty IOP elevation, in 35% of subjects with keratoconus, is markedly higher in this New Zealand study than previously reported in the US and UK studies. Further clinical and genetic analysis of associations between keratoconus and steroid-induced IOP elevation and glaucoma might improve our current understanding of this condition. PMID- 19300465 TI - Red reflex mimicking lens subluxation in a child undergoing orthokeratology. PMID- 19300466 TI - Unilateral ptosis correction with mersilene mesh frontalis sling in infants: thirteen-year follow-up report. AB - PURPOSE: To assess surgical, visual, refractive, and aesthetic outcomes 13 years after mersilene mesh frontalis sling (MMFS) operation for severe unilateral congenital ptosis performed in 10 infants before 1 year of age. METHODS: Longitudinal follow-up of an interventional case series by structured ocular examinations, external photographs, and questionnaire-based interviews. RESULTS: Mean age at surgery was 6.9+/-2.7 months. After a mean follow-up of 13.0+/-0.6 years, one patient (10%) had recurrent ptosis with the upper lid 2 mm below the superior limbus at 3 months postoperatively. Best-corrected visual acuities were within two Snellen lines between the two eyes in all patients. Astigmatic errors were 1.20+/-1.00 D and 1.10+/-1.70 D between operated and unoperated eyes. Four patients had 2 mm lid lag on down-gaze and one of them had 2 mm lagophthalmos. Mean satisfaction scores (scale of 1 to 100) for lid position, cosmesis, function, and to the procedure were 83.3+/-11.8, 77.0+/-22.9, 89.4+/-5.5, and 86.8+/-6.3, respectively. No case of overcorrection, sling extrusion, stitch granuloma, or exposure keratopathy was noted. CONCLUSIONS: In view of the low recurrence rate (10%) and absence of serious complication or need for revision after 13 years, the use of MMFS seems effective and feasible in infants less than 1 year old. Achieving compatible long-term stability, satisfactory aesthetic, and visual outcomes, MMFS may offer an alternative to delaying operations for autogenous fascia lata harvesting in infants requiring early ptosis correction. PMID- 19300467 TI - Severe allergic blepharoconjunctivitis after eyelash colouring. PMID- 19300468 TI - JBIR-17, a novel trichostatin analog from Streptomyces sp. 26634. PMID- 19300469 TI - Isolation, structure elucidation and antibacterial activities of streptothricin acids. AB - Five streptothricin acids (compounds 1-5) were isolated by ion-exchange resin chromatography and preparative RP-HPLC from the fermentation broth of Streptomyces qinlingensis. Their structures were elucidated mainly by analyses of the IR, HR-EIS-MS and NMR spectral data. They were deduced as two known compounds, streptothricin F acid (1) and streptothricin D acid (2), and three new compounds, 12-carbamoylstreptothricin E acid (3), 12-carbamoylstreptothricin D acid (4) and N-acetyl-streptothricin D acid (5), respectively. The antibacterial activities of 1-5 against Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Bacillus cereus and Pseudomonas aeruginosa were assayed by micro-broth dilution. Comparison of the MICs with streptothricin F and D showed that the antimicrobial activities of 1-5 were decreased significantly but varied with the structures. PMID- 19300470 TI - Pentacecilides, new inhibitors of lipid droplet formation in mouse macrophages, produced by Penicillium cecidicola FKI-3765-1: I. Taxonomy, fermentation, isolation and biological properties. AB - New compounds designated pentacecilides A to C were isolated from the fermentation broth of Penicillium cecidicola FKI-3765-1 by solvent extraction, silica gel column chromatography and preparative HPLC. Pentacecilides A and B dose-dependently inhibited lipid droplet formation in mouse macrophages. Furthermore, pentacecilides A and B were found to inhibit the synthesis of cholesteryl ester in mouse macrophages with respective IC(50) values of 3.65 and 4.76 microM without any cytotoxic effect, but pentacecilide C showed almost no activity. The study of the mechanism of action strongly suggested that pentacecilides A and B inhibit acyl-CoA: cholesterol acyltransferase activity in macrophages. PMID- 19300471 TI - Pentacecilides, new inhibitors of lipid droplet formation in mouse macrophages produced by Penicillium cecidicola FKI-3765-1: II. Structure elucidation. AB - The structures of pentacecilides, new inhibitors of lipid droplet formation in mouse macrophages produced by Penicillium cecidicola FKI-3765-1, were elucidated by spectroscopic studies, including various NMR experiments. Pentacecilides have a common pentacyclic meroterpene core, which contains an aromatic ring and a delta-lactone ring. PMID- 19300472 TI - MLH1 deficiency enhances tumor cell sensitivity to ganciclovir. AB - Suicide gene therapy with herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir (GCV) is notable for producing multi-log cytotoxicity in a unique pattern of delayed cytotoxicity in S-phase. As hydroxyurea, a ribonucleotide reductase inhibitor that activates mismatch repair, can increase sensitivity to GCV, we evaluated the role of MLH1, an essential mismatch repair protein, in GCV cytotoxicity. Using HCT116TK (HSV-TK-expressing) colon carcinoma cells that express or lack MLH1, cell-survival studies demonstrated greater GCV sensitivity in the MLH1-deficient cells, primarily at high concentrations. This could not be explained by differences in GCV metabolism, as the less sensitive MLH1 expresssing cells accumulated more GCV triphosphate and incorporated more of the analog into DNA. SiRNA suppression of MLH1 in U251 glioblastoma or SW480 colon carcinoma cells also enhanced sensitivity to high concentrations of GCV. Studies in a pa nel of yeast deletion mutants confirmed the results with MLH1, and further suggested a role for homologous recombination repair and several cell cycle checkpoint proteins in GCV cytotoxicity. These data suggest that MLH1 can prevent cytotoxicity with GCV. Targeting mismatch repair-deficient tumors may increase efficacy of this suicide gene therapy approach to cancer treatment. PMID- 19300473 TI - Broadband criticality of human brain network synchronization. AB - Self-organized criticality is an attractive model for human brain dynamics, but there has been little direct evidence for its existence in large-scale systems measured by neuroimaging. In general, critical systems are associated with fractal or power law scaling, long-range correlations in space and time, and rapid reconfiguration in response to external inputs. Here, we consider two measures of phase synchronization: the phase-lock interval, or duration of coupling between a pair of (neurophysiological) processes, and the lability of global synchronization of a (brain functional) network. Using computational simulations of two mechanistically distinct systems displaying complex dynamics, the Ising model and the Kuramoto model, we show that both synchronization metrics have power law probability distributions specifically when these systems are in a critical state. We then demonstrate power law scaling of both pairwise and global synchronization metrics in functional MRI and magnetoencephalographic data recorded from normal volunteers under resting conditions. These results strongly suggest that human brain functional systems exist in an endogenous state of dynamical criticality, characterized by a greater than random probability of both prolonged periods of phase-locking and occurrence of large rapid changes in the state of global synchronization, analogous to the neuronal "avalanches" previously described in cellular systems. Moreover, evidence for critical dynamics was identified consistently in neurophysiological systems operating at frequency intervals ranging from 0.05-0.11 to 62.5-125 Hz, confirming that criticality is a property of human brain functional network organization at all frequency intervals in the brain's physiological bandwidth. PMID- 19300474 TI - Computationally driven, quantitative experiments discover genes required for mitochondrial biogenesis. AB - Mitochondria are central to many cellular processes including respiration, ion homeostasis, and apoptosis. Using computational predictions combined with traditional quantitative experiments, we have identified 100 proteins whose deficiency alters mitochondrial biogenesis and inheritance in Saccharomyces cerevisiae. In addition, we used computational predictions to perform targeted double-mutant analysis detecting another nine genes with synthetic defects in mitochondrial biogenesis. This represents an increase of about 25% over previously known participants. Nearly half of these newly characterized proteins are conserved in mammals, including several orthologs known to be involved in human disease. Mutations in many of these genes demonstrate statistically significant mitochondrial transmission phenotypes more subtle than could be detected by traditional genetic screens or high-throughput techniques, and 47 have not been previously localized to mitochondria. We further characterized a subset of these genes using growth profiling and dual immunofluorescence, which identified genes specifically required for aerobic respiration and an uncharacterized cytoplasmic protein required for normal mitochondrial motility. Our results demonstrate that by leveraging computational analysis to direct quantitative experimental assays, we have characterized mutants with subtle mitochondrial defects whose phenotypes were undetected by high-throughput methods. PMID- 19300475 TI - Using network component analysis to dissect regulatory networks mediated by transcription factors in yeast. AB - Understanding the relationship between genetic variation and gene expression is a central question in genetics. With the availability of data from high-throughput technologies such as ChIP-Chip, expression, and genotyping arrays, we can begin to not only identify associations but to understand how genetic variations perturb the underlying transcription regulatory networks to induce differential gene expression. In this study, we describe a simple model of transcription regulation where the expression of a gene is completely characterized by two properties: the concentrations and promoter affinities of active transcription factors. We devise a method that extends Network Component Analysis (NCA) to determine how genetic variations in the form of single nucleotide polymorphisms (SNPs) perturb these two properties. Applying our method to a segregating population of Saccharomyces cerevisiae, we found statistically significant examples of trans-acting SNPs located in regulatory hotspots that perturb transcription factor concentrations and affinities for target promoters to cause global differential expression and cis-acting genetic variations that perturb the promoter affinities of transcription factors on a single gene to cause local differential expression. Although many genetic variations linked to gene expressions have been identified, it is not clear how they perturb the underlying regulatory networks that govern gene expression. Our work begins to fill this void by showing that many genetic variations affect the concentrations of active transcription factors in a cell and their affinities for target promoters. Understanding the effects of these perturbations can help us to paint a more complete picture of the complex landscape of transcription regulation. The software package implementing the algorithms discussed in this work is available as a MATLAB package upon request. PMID- 19300476 TI - On the growth of scientific knowledge: yeast biology as a case study. AB - The tempo and mode of human knowledge expansion is an enduring yet poorly understood topic. Through a temporal network analysis of three decades of discoveries of protein interactions and genetic interactions in baker's yeast, we show that the growth of scientific knowledge is exponential over time and that important subjects tend to be studied earlier. However, expansions of different domains of knowledge are highly heterogeneous and episodic such that the temporal turnover of knowledge hubs is much greater than expected by chance. Familiar subjects are preferentially studied over new subjects, leading to a reduced pace of innovation. While research is increasingly done in teams, the number of discoveries per researcher is greater in smaller teams. These findings reveal collective human behaviors in scientific research and help design better strategies in future knowledge exploration. PMID- 19300477 TI - Sfrp controls apicobasal polarity and oriented cell division in developing gut epithelium. AB - Epithelial tubular morphogenesis leading to alteration of organ shape has important physiological consequences. However, little is known regarding the mechanisms that govern epithelial tube morphogenesis. Here, we show that inactivation of Sfrp1 and Sfrp2 leads to reduction in fore-stomach length in mouse embryos, which is enhanced in the presence of the Sfrp5 mutation. In the mono-cell layer of fore-stomach epithelium, cell division is normally oriented along the cephalocaudal axis; in contrast, orientation diverges in the Sfrps deficient fore-stomach. Cell growth and apoptosis are not affected in the Sfrps deficient fore-stomach epithelium. Similarly, cell division orientation in fore stomach epithelium diverges as a result of inactivation of either Stbm/Vangl2, an Fz/PCP component, or Wnt5a. These observations indicate that the oriented cell division, which is controlled by the Fz/PCP pathway, is one of essential components in fore-stomach morphogenesis. Additionally, the small intestine epithelium of Sfrps compound mutants fails to maintain proper apicobasal polarity; the defect was also observed in Wnt5a-inactivated small intestine. In relation to these findings, Sfrp1 physically interacts with Wnt5a and inhibits Wnt5a signaling. We propose that Sfrp regulation of Wnt5a signaling controls oriented cell division and apicobasal polarity in the epithelium of developing gut. PMID- 19300478 TI - Modeling signal propagation mechanisms and ligand-based conformational dynamics of the Hsp90 molecular chaperone full-length dimer. AB - Hsp90 is a molecular chaperone essential for protein folding and activation in normal homeostasis and stress response. ATP binding and hydrolysis facilitate Hsp90 conformational changes required for client activation. Hsp90 plays an important role in disease states, particularly in cancer, where chaperoning of the mutated and overexpressed oncoproteins is important for function. Recent studies have illuminated mechanisms related to the chaperone function. However, an atomic resolution view of Hsp90 conformational dynamics, determined by the presence of different binding partners, is critical to define communication pathways between remote residues in different domains intimately affecting the chaperone cycle. Here, we present a computational analysis of signal propagation and long-range communication pathways in Hsp90. We carried out molecular dynamics simulations of the full-length Hsp90 dimer, combined with essential dynamics, correlation analysis, and a signal propagation model. All-atom MD simulations with timescales of 70 ns have been performed for complexes with the natural substrates ATP and ADP and for the unliganded dimer. We elucidate the mechanisms of signal propagation and determine "hot spots" involved in interdomain communication pathways from the nucleotide-binding site to the C-terminal domain interface. A comprehensive computational analysis of the Hsp90 communication pathways and dynamics at atomic resolution has revealed the role of the nucleotide in effecting conformational changes, elucidating the mechanisms of signal propagation. Functionally important residues and secondary structure elements emerge as effective mediators of communication between the nucleotide binding site and the C-terminal interface. Furthermore, we show that specific interdomain signal propagation pathways may be activated as a function of the ligand. Our results support a "conformational selection model" of the Hsp90 mechanism, whereby the protein may exist in a dynamic equilibrium between different conformational states available on the energy landscape and binding of a specific partner can bias the equilibrium toward functionally relevant complexes. PMID- 19300479 TI - Computational model of the insect pheromone transduction cascade. AB - A biophysical model of receptor potential generation in the male moth olfactory receptor neuron is presented. It takes into account all pre-effector processes- the translocation of pheromone molecules from air to sensillum lymph, their deactivation and interaction with the receptors, and the G-protein and effector enzyme activation--and focuses on the main post-effector processes. These processes involve the production and degradation of second messengers (IP(3) and DAG), the opening and closing of a series of ionic channels (IP(3)-gated Ca(2+) channel, DAG-gated cationic channel, Ca(2+)-gated Cl(-) channel, and Ca(2+)- and voltage-gated K(+) channel), and Ca(2+) extrusion mechanisms. The whole network is regulated by modulators (protein kinase C and Ca(2+)-calmodulin) that exert feedback inhibition on the effector and channels. The evolution in time of these linked chemical species and currents and the resulting membrane potentials in response to single pulse stimulation of various intensities were simulated. The unknown parameter values were fitted by comparison to the amplitude and temporal characteristics (rising and falling times) of the experimentally measured receptor potential at various pheromone doses. The model obtained captures the main features of the dose-response curves: the wide dynamic range of six decades with the same amplitudes as the experimental data, the short rising time, and the long falling time. It also reproduces the second messenger kinetics. It suggests that the two main types of depolarizing ionic channels play different roles at low and high pheromone concentrations; the DAG-gated cationic channel plays the major role for depolarization at low concentrations, and the Ca(2+)-gated Cl(-) channel plays the major role for depolarization at middle and high concentrations. Several testable predictions are proposed, and future developments are discussed. PMID- 19300480 TI - Germline mutation in NLRP2 (NALP2) in a familial imprinting disorder (Beckwith Wiedemann Syndrome). AB - Beckwith-Wiedemann syndrome (BWS) is a fetal overgrowth and human imprinting disorder resulting from the deregulation of a number of genes, including IGF2 and CDKN1C, in the imprinted gene cluster on chromosome 11p15.5. Most cases are sporadic and result from epimutations at either of the two 11p15.5 imprinting centres (IC1 and IC2). However, rare familial cases may be associated with germline 11p15.5 deletions causing abnormal imprinting in cis. We report a family with BWS and an IC2 epimutation in which affected siblings had inherited different parental 11p15.5 alleles excluding an in cis mechanism. Using a positional-candidate gene approach, we found that the mother was homozygous for a frameshift mutation in exon 6 of NLRP2. While germline mutations in NLRP7 have previously been associated with familial hydatidiform mole, this is the first description of NLRP2 mutation in human disease and the first report of a trans mechanism for disordered imprinting in BWS. These observations are consistent with the hypothesis that NLRP2 has a previously unrecognised role in establishing or maintaining genomic imprinting in humans. PMID- 19300481 TI - Ciliary beating recovery in deficient human airway epithelial cells after lentivirus ex vivo gene therapy. AB - Primary Ciliary Dyskinesia is a heterogeneous genetic disease that is characterized by cilia dysfunction of the epithelial cells lining the respiratory tracts, resulting in recurrent respiratory tract infections. Despite lifelong physiological therapy and antibiotics, the lungs of affected patients are progressively destroyed, leading to respiratory insufficiency. Recessive mutations in Dynein Axonemal Intermediate chain type 1 (DNAI1) gene have been described in 10% of cases of Primary Ciliary Dyskinesia. Our goal was to restore normal ciliary beating in DNAI1-deficient human airway epithelial cells. A lentiviral vector based on Simian Immunodeficiency Virus pseudotyped with Vesicular Stomatitis Virus Glycoprotein was used to transduce cultured human airway epithelial cells with a cDNA of DNAI1 driven by the Elongation Factor 1 promoter. Transcription and translation of the transduced gene were tested by RT PCR and western blot, respectively. Human airway epithelial cells that were DNAI1 deficient due to compound heterozygous mutations, and consequently had immotile cilia and no outer dynein arm, were transduced by the lentivirus. Cilia beating was recorded and electron microscopy of the cilia was performed. Transcription and translation of the transduced DNAI1 gene were detected in human cells treated with the lentivirus. In addition, immotile cilia recovered a normal beat and outer dynein arms reappeared. We demonstrated that it is possible to obtain a normalization of ciliary beat frequency of deficient human airway epithelial cells by using a lentivirus to transduce cells with the therapeutic gene. This preliminary step constitutes a conceptual proof that is indispensable in the perspective of Primary Ciliary Dyskinesia's in vivo gene therapy. This is the first time that recovery of cilia beating is demonstrated in this disease. PMID- 19300482 TI - A genome-wide association study in chronic obstructive pulmonary disease (COPD): identification of two major susceptibility loci. AB - There is considerable variability in the susceptibility of smokers to develop chronic obstructive pulmonary disease (COPD). The only known genetic risk factor is severe deficiency of alpha(1)-antitrypsin, which is present in 1-2% of individuals with COPD. We conducted a genome-wide association study (GWAS) in a homogenous case-control cohort from Bergen, Norway (823 COPD cases and 810 smoking controls) and evaluated the top 100 single nucleotide polymorphisms (SNPs) in the family-based International COPD Genetics Network (ICGN; 1891 Caucasian individuals from 606 pedigrees) study. The polymorphisms that showed replication were further evaluated in 389 subjects from the US National Emphysema Treatment Trial (NETT) and 472 controls from the Normative Aging Study (NAS) and then in a fourth cohort of 949 individuals from 127 extended pedigrees from the Boston Early-Onset COPD population. Logistic regression models with adjustments of covariates were used to analyze the case-control populations. Family-based association analyses were conducted for a diagnosis of COPD and lung function in the family populations. Two SNPs at the alpha-nicotinic acetylcholine receptor (CHRNA 3/5) locus were identified in the genome-wide association study. They showed unambiguous replication in the ICGN family-based analysis and in the NETT case-control analysis with combined p-values of 1.48 x 10(-10), (rs8034191) and 5.74 x 10(-10) (rs1051730). Furthermore, these SNPs were significantly associated with lung function in both the ICGN and Boston Early-Onset COPD populations. The C allele of the rs8034191 SNP was estimated to have a population attributable risk for COPD of 12.2%. The association of hedgehog interacting protein (HHIP) locus on chromosome 4 was also consistently replicated, but did not reach genome wide significance levels. Genome-wide significant association of the HHIP locus with lung function was identified in the Framingham Heart study (Wilk et al., companion article in this issue of PLoS Genetics; doi:10.1371/journal.pgen.1000429). The CHRNA 3/5 and the HHIP loci make a significant contribution to the risk of COPD. CHRNA3/5 is the same locus that has been implicated in the risk of lung cancer. PMID- 19300483 TI - Quantifying global tolerance of biochemical systems: design implications for moiety-transfer cycles. AB - Robustness of organisms is widely observed although difficult to precisely characterize. Performance can remain nearly constant within some neighborhood of the normal operating regime, leading to homeostasis, but then abruptly break down with pathological consequences beyond this neighborhood. Currently, there is no generic approach to identifying boundaries where local performance deteriorates abruptly, and this has hampered understanding of the molecular basis of biological robustness. Here we introduce a generic approach for characterizing boundaries between operational regimes based on the piecewise power-law representation of the system's components. This conceptual framework allows us to define "global tolerance" as the ratio between the normal value of a parameter and the value at such a boundary. We illustrate the utility of this concept for a class of moiety-transfer cycles, which is a widespread module in biology. Our results show a region of "best" local performance surrounded by "poor" regions; also, selection for improved local performance often pushes the operating values away from regime boundaries, thus increasing global tolerance. These predictions agree with experimental data from the reduced nicotinamide adenine dinucleotide phosphate (NADPH) redox cycle of human erythrocytes. PMID- 19300484 TI - T-cell epitope prediction: rescaling can mask biological variation between MHC molecules. AB - Theoretical methods for predicting CD8+ T-cell epitopes are an important tool in vaccine design and for enhancing our understanding of the cellular immune system. The most popular methods currently available produce binding affinity predictions across a range of MHC molecules. In comparing results between these MHC molecules, it is common practice to apply a normalization procedure known as rescaling, to correct for possible discrepancies between the allelic predictors. Using two of the most popular prediction software packages, NetCTL and NetMHC, we tested the hypothesis that rescaling removes genuine biological variation from the predicted affinities when comparing predictions across a number of MHC molecules. We found that removing the condition of rescaling improved the prediction software's performance both qualitatively, in terms of ranking epitopes, and quantitatively, in the accuracy of their binding affinity predictions. We suggest that there is biologically significant variation among class 1 MHC molecules and find that retention of this variation leads to significantly more accurate epitope prediction. PMID- 19300485 TI - Independent S-locus mutations caused self-fertility in Arabidopsis thaliana. AB - A common yet poorly understood evolutionary transition among flowering plants is a switch from outbreeding to an inbreeding mode of mating. The model plant Arabidopsis thaliana evolved to an inbreeding state through the loss of self incompatibility, a pollen-rejection system in which pollen recognition by the stigma is determined by tightly linked and co-evolving alleles of the S-locus receptor kinase (SRK) and its S-locus cysteine-rich ligand (SCR). Transformation of A. thaliana, with a functional AlSRKb-SCRb gene pair from its outcrossing relative A. lyrata, demonstrated that A. thaliana accessions harbor different sets of cryptic self-fertility-promoting mutations, not only in S-locus genes, but also in other loci required for self-incompatibility. However, it is still not known how many times and in what manner the switch to self-fertility occurred in the A. thaliana lineage. Here, we report on our identification of four accessions that are reverted to full self-incompatibility by transformation with AlSRKb-SCRb, bringing to five the number of accessions in which self-fertility is due to, and was likely caused by, S-locus inactivation. Analysis of S-haplotype organization reveals that inter-haplotypic recombination events, rearrangements, and deletions have restructured the S locus and its genes in these accessions. We also perform a Quantitative Trait Loci (QTL) analysis to identify modifier loci associated with self-fertility in the Col-0 reference accession, which cannot be reverted to full self-incompatibility. Our results indicate that the transition to inbreeding occurred by at least two, and possibly more, independent S-locus mutations, and identify a novel unstable modifier locus that contributes to self fertility in Col-0. PMID- 19300486 TI - Evolution of a bacterial regulon controlling virulence and Mg(2+) homeostasis. AB - Related organisms typically rely on orthologous regulatory proteins to respond to a given signal. However, the extent to which (or even if) the targets of shared regulatory proteins are maintained across species has remained largely unknown. This question is of particular significance in bacteria due to the widespread effects of horizontal gene transfer. Here, we address this question by investigating the regulons controlled by the DNA-binding PhoP protein, which governs virulence and Mg(2+) homeostasis in several bacterial species. We establish that the ancestral PhoP protein directs largely different gene sets in ten analyzed species of the family Enterobacteriaceae, reflecting both regulation of species-specific targets and transcriptional rewiring of shared genes. The two targets directly activated by PhoP in all ten species (the most distant of which diverged >200 million years ago), and coding for the most conserved proteins are the phoPQ operon itself and the lipoprotein-encoding slyB gene, which decreases PhoP protein activity. The Mg(2+)-responsive PhoP protein dictates expression of Mg(2+) transporters and of enzymes that modify Mg(2+)-binding sites in the cell envelope in most analyzed species. In contrast to the core PhoP regulon, which determines the amount of active PhoP and copes with the low Mg(2+) stress, the variable members of the regulon contribute species-specific traits, a property shared with regulons controlled by dissimilar regulatory proteins and responding to different signals. PMID- 19300487 TI - Accurate detection of recombinant breakpoints in whole-genome alignments. AB - We propose a novel method for detecting sites of molecular recombination in multiple alignments. Our approach is a compromise between previous extremes of computationally prohibitive but mathematically rigorous methods and imprecise heuristic methods. Using a combined algorithm for estimating tree structure and hidden Markov model parameters, our program detects changes in phylogenetic tree topology over a multiple sequence alignment. We evaluate our method on benchmark datasets from previous studies on two recombinant pathogens, Neisseria and HIV-1, as well as simulated data. We show that we are not only able to detect recombinant regions of vastly different sizes but also the location of breakpoints with great accuracy. We show that our method does well inferring recombination breakpoints while at the same time maintaining practicality for larger datasets. In all cases, we confirm the breakpoint predictions of previous studies, and in many cases we offer novel predictions. PMID- 19300488 TI - Parallel germline infiltration of a lentivirus in two Malagasy lemurs. AB - Retroviruses normally infect the somatic cells of their host and are transmitted horizontally, i.e., in an exogenous way. Occasionally, however, some retroviruses can also infect and integrate into the genome of germ cells, which may allow for their vertical inheritance and fixation in a given species; a process known as endogenization. Lentiviruses, a group of mammalian retroviruses that includes HIV, are known to infect primates, ruminants, horses, and cats. Unlike many other retroviruses, these viruses have not been demonstrably successful at germline infiltration. Here, we report on the discovery of endogenous lentiviral insertions in seven species of Malagasy lemurs from two different genera -- Cheirogaleus and Microcebus. Combining molecular clock analyses and cross-species screening of orthologous insertions, we show that the presence of this endogenous lentivirus in six species of Microcebus is the result of one endogenization event that occurred about 4.2 million years ago. In addition, we demonstrate that this lentivirus independently infiltrated the germline of Cheirogaleus and that the two endogenization events occurred quasi-simultaneously. Using multiple proviral copies, we derive and characterize an apparently full length and intact consensus for this lentivirus. These results provide evidence that lentiviruses have repeatedly infiltrated the germline of prosimian species and that primates have been exposed to lentiviruses for a much longer time than what can be inferred based on sequence comparison of circulating lentiviruses. The study sets the stage for an unprecedented opportunity to reconstruct an ancestral primate lentivirus and thereby advance our knowledge of host-virus interactions. PMID- 19300489 TI - Ras conformational switching: simulating nucleotide-dependent conformational transitions with accelerated molecular dynamics. AB - Ras mediates signaling pathways controlling cell proliferation and development by cycling between GTP- and GDP-bound active and inactive conformational states. Understanding the complete reaction path of this conformational change and its intermediary structures is critical to understanding Ras signaling. We characterize nucleotide-dependent conformational transition using multiple barrier-crossing accelerated molecular dynamics (aMD) simulations. These transitions, achieved for the first time for wild-type Ras, are impossible to observe with classical molecular dynamics (cMD) simulations due to the large energetic barrier between end states. Mapping the reaction path onto a conformer plot describing the distribution of the crystallographic structures enabled identification of highly populated intermediate structures. These structures have unique switch orientations (residues 25-40 and 57-75) intermediate between GTP and GDP states, or distinct loop3 (46-49), loop7 (105-110), and alpha5 C-terminus (159-166) conformations distal from the nucleotide-binding site. In addition, these barrier-crossing trajectories predict novel nucleotide-dependent correlated motions, including correlations of alpha2 (residues 66-74) with alpha3-loop7 (93 110), loop2 (26-37) with loop10 (145-151), and loop3 (46-49) with alpha5 (152 167). The interconversion between newly identified Ras conformations revealed by this study advances our mechanistic understanding of Ras function. In addition, the pattern of correlated motions provides new evidence for a dynamic linkage between the nucleotide-binding site and the membrane interacting C-terminus critical for the signaling function of Ras. Furthermore, normal mode analysis indicates that the dominant collective motion that occurs during nucleotide dependent conformational exchange, and captured in aMD (but absent in cMD) simulations, is a low-frequency motion intrinsic to the structure. PMID- 19300490 TI - An African ancestry-specific allele of CTLA4 confers protection against rheumatoid arthritis in African Americans. AB - Cytotoxic T-lymphocyte associated protein 4 (CTLA4) is a negative regulator of T cell proliferation. Polymorphisms in CTLA4 have been inconsistently associated with susceptibility to rheumatoid arthritis (RA) in populations of European ancestry but have not been examined in African Americans. The prevalence of RA in most populations of European and Asian ancestry is approximately 1.0%; RA is purportedly less common in black Africans, with little known about its prevalence in African Americans. We sought to determine if CTLA4 polymorphisms are associated with RA in African Americans. We performed a 2-stage analysis of 12 haplotype tagging single nucleotide polymorphisms (SNPs) across CTLA4 in a total of 505 African American RA patients and 712 African American controls using Illumina and TaqMan platforms. The minor allele (G) of the rs231778 SNP was 0.054 in RA patients, compared to 0.209 in controls (4.462 x 10(-26), Fisher's exact). The presence of the G allele was associated with a substantially reduced odds ratio (OR) of having RA (AG+GG genotypes vs. AA genotype, OR 0.19, 95% CI: 0.13 0.26, p = 2.4 x 10(-28), Fisher's exact), suggesting a protective effect. This SNP is polymorphic in the African population (minor allele frequency [MAF] 0.09 in the Yoruba population), but is very rare in other groups (MAF = 0.002 in 530 Caucasians genotyped for this study). Markers associated with RA in populations of European ancestry (rs3087243 [+60C/T] and rs231775 [+49A/G]) were not replicated in African Americans. We found no confounding of association for rs231778 after stratifying for the HLA-DRB1 shared epitope, presence of anti cyclic citrullinated peptide antibody, or degree of admixture from the European population. An African ancestry-specific genetic variant of CTLA4 appears to be associated with protection from RA in African Americans. This finding may explain, in part, the relatively low prevalence of RA in black African populations. PMID- 19300491 TI - Gag mutations strongly contribute to HIV-1 resistance to protease inhibitors in highly drug-experienced patients besides compensating for fitness loss. AB - Human immunodeficiency virus type 1 (HIV-1) resistance to protease inhibitors (PI) results from mutations in the viral protease (PR) that reduce PI binding but also decrease viral replicative capacity (RC). Additional mutations compensating for the RC loss subsequently accumulate within PR and in Gag substrate cleavage sites. We examined the respective contribution of mutations in PR and Gag to PI resistance and RC and their interdependence using a panel of HIV-1 molecular clones carrying different sequences from six patients who had failed multiple lines of treatment. Mutations in Gag strongly and directly contributed to PI resistance besides compensating for fitness loss. This effect was essentially carried by the C-terminal region of Gag (containing NC-SP2-p6) with little or no contribution from MA, CA, and SP1. The effect of Gag on resistance depended on the presence of cleavage site mutations A431V or I437V in NC-SP2-p6 and correlated with processing of the NC/SP2 cleavage site. By contrast, reverting the A431V or I437V mutation in these highly evolved sequences had little effect on RC. Mutations in the NC-SP2-p6 region of Gag can be dually selected as compensatory and as direct PI resistance mutations, with cleavage at the NC-SP2 site behaving as a rate-limiting step in PI resistance. Further compensatory mutations render viral RC independent of the A431V or I437V mutations while their effect on resistance persists. PMID- 19300492 TI - Activation of host translational control pathways by a viral developmental switch. AB - In response to numerous signals, latent herpesvirus genomes abruptly switch their developmental program, aborting stable host-cell colonization in favor of productive viral replication that ultimately destroys the cell. To achieve a rapid gene expression transition, newly minted capped, polyadenylated viral mRNAs must engage and reprogram the cellular translational apparatus. While transcriptional responses of viral genomes undergoing lytic reactivation have been amply documented, roles for cellular translational control pathways in enabling the latent-lytic switch have not been described. Using PEL-derived B cells naturally infected with KSHV as a model, we define efficient reactivation conditions and demonstrate that reactivation substantially changes the protein synthesis profile. New polypeptide synthesis correlates with 4E-BP1 translational repressor inactivation, nuclear PABP accumulation, eIF4F assembly, and phosphorylation of the cap-binding protein eIF4E by Mnk1. Significantly, inhibiting Mnk1 reduces accumulation of the critical viral transactivator RTA through a post-transcriptional mechanism, limiting downstream lytic protein production, and impairs reactivation efficiency. Thus, herpesvirus reactivation from latency activates the host cap-dependent translation machinery, illustrating the importance of translational regulation in implementing new developmental instructions that drastically alter cell fate. PMID- 19300493 TI - Severe Plasmodium falciparum malaria is associated with circulating ultra-large von Willebrand multimers and ADAMTS13 inhibition. AB - Plasmodium falciparum infection results in adhesion of infected erythrocytes to blood vessel endothelium, and acute endothelial cell activation, together with sequestration of platelets and leucocytes. We have previously shown that patients with severe infection or fulminant cerebral malaria have significantly increased circulatory levels of the adhesive glycoprotein von Willebrand factor (VWF) and its propeptide, both of which are indices of endothelial cell activation. In this prospective study of patients from Ghana with severe (n = 20) and cerebral (n = 13) P. falciparum malaria, we demonstrate that increased plasma VWF antigen (VWF:Ag) level is associated with disproportionately increased VWF function. VWF collagen binding (VWF:CB) was significantly increased in patients with cerebral malaria and severe malaria (medians 7.6 and 7.0 IU/ml versus 1.9 IU/ml; p<0.005). This increased VWF:CB correlated with the presence of abnormal ultra-large VWF multimers in patient rather than control plasmas. Concomitant with the increase in VWF:Ag and VWF:CB was a significant persistent reduction in the activity of the VWF-specific cleaving protease ADAMTS13 (approximately 55% of normal; p<0.005). Mixing studies were performed using P. falciparum patient plasma and normal pooled plasma, in the presence or absence of exogenous recombinant ADAMTS13. These studies demonstrated that in malarial plasma, ADAMTS13 function was persistently inhibited in a time-dependent manner. Furthermore, this inhibitory effect was not associated with the presence of known inhibitors of ADAMTS13 enzymatic function (interleukin-6, free haemoglobin, factor VIII or thrombospondin-1). These novel findings suggest that severe P. falciparum infection is associated with acute endothelial cell activation, abnormal circulating ULVWF multimers, and a significant reduction in plasma ADAMTS13 function which is mediated at least in part by an unidentified inhibitor. PMID- 19300494 TI - A systems approach uncovers restrictions for signal interactions regulating genome-wide responses to nutritional cues in Arabidopsis. AB - As sessile organisms, plants must cope with multiple and combined variations of signals in their environment. However, very few reports have studied the genome wide effects of systematic signal combinations on gene expression. Here, we evaluate a high level of signal integration, by modeling genome-wide expression patterns under a factorial combination of carbon (C), light (L), and nitrogen (N) as binary factors in two organs (O), roots and leaves. Signal management is different between C, N, and L and in shoots and roots. For example, L is the major factor controlling gene expression in leaves. However, in roots there is no obvious prominent signal, and signal interaction is stronger. The major signal interaction events detected genome wide in Arabidopsis roots are deciphered and summarized in a comprehensive conceptual model. Surprisingly, global analysis of gene expression in response to C, N, L, and O revealed that the number of genes controlled by a signal is proportional to the magnitude of the gene expression changes elicited by the signal. These results uncovered a strong constraining structure in plant cell signaling pathways, which prompted us to propose the existence of a "code" of signal integration. PMID- 19300495 TI - Reassessing the role of APOBEC3G in human immunodeficiency virus type 1 infection of quiescent CD4+ T-cells. AB - HIV-1 is restricted for infection of primary quiescent T-cells. After viral entry, reverse transcription is initiated but is not completed. Various hypotheses have been proposed for this cellular restriction including insufficient nucleotide pools and cellular factors, but none have been confirmed as the primary mechanism for restriction. A recent study by Chiu et al. implicates APOBEC3G, an anti-retroviral cytidine deaminase, as the cellular restriction factor. Here, we attempted to confirm these findings using the same strategy as reported by Chiu et al. of siRNA targeting knock-down of APOBEC3G expression. In contrast to the published study, our results do not support a role for APOBEC3G in restriction of HIV-1 in quiescent CD4+ T-cells. In our study, we tested the same siRNA as reported by Chiu et al. as well as two additional siRNAs targeting APOBEC3G, one of which showed 2-fold greater knock-down of APOBEC3G mRNA. However, none of the three siRNAs tested had a discernable effect on enhancing infection by HIV-1 in quiescent CD4+ T-cells. Therefore, we conclude that the primary mechanism of HIV-1 restriction in quiescent CD4+ T-cells remains to be elucidated. PMID- 19300496 TI - Wolbachia-mediated cytoplasmic incompatibility is associated with impaired histone deposition in the male pronucleus. AB - Wolbachia is a bacteria endosymbiont that rapidly infects insect populations through a mechanism known as cytoplasmic incompatibility (CI). In CI, crosses between Wolbachia-infected males and uninfected females produce severe cell cycle defects in the male pronucleus resulting in early embryonic lethality. In contrast, viable progeny are produced when both parents are infected (the Rescue cross). An important consequence of CI-Rescue is that infected females have a selective advantage over uninfected females facilitating the rapid spread of Wolbachia through insect populations. CI disrupts a number of prophase and metaphase events in the male pronucleus, including Cdk1 activation, chromosome condensation, and segregation. Here, we demonstrate that CI disrupts earlier interphase cell cycle events. Specifically, CI delays the H3.3 and H4 deposition that occurs immediately after protamine removal from the male pronucleus. In addition, we find prolonged retention of the replication factor PCNA in the male pronucleus into metaphase, indicating progression into mitosis with incompletely replicated DNA. We propose that these CI-induced interphase defects in de novo nucleosome assembly and replication are the cause of the observed mitotic condensation and segregation defects. In addition, these interphase chromosome defects likely activate S-phase checkpoints, accounting for the previously described delays in Cdk1 activation. These results have important implications for the mechanism of Rescue and other Wolbachia-induced phenotypes. PMID- 19300497 TI - Cross-protective potential of a novel monoclonal antibody directed against antigenic site B of the hemagglutinin of influenza A viruses. AB - The hemagglutinin (HA) of influenza A viruses has been classified into sixteen distinct subtypes (H1-H16) to date. The HA subtypes of influenza A viruses are principally defined as serotypes determined by neutralization or hemagglutination inhibition tests using polyclonal antisera to the respective HA subtypes, which have little cross-reactivity to the other HA subtypes. Thus, it is generally believed that the neutralizing antibodies are not broadly cross-reactive among HA subtypes. In this study, we generated a novel monoclonal antibody (MAb) specific to HA, designated MAb S139/1, which showed heterosubtypic cross-reactive neutralization and hemagglutination inhibition of influenza A viruses. This MAb was found to have broad reactivity to many other viruses (H1, H2, H3, H5, H9, and H13 subtypes) in enzyme-linked immunosorbent assays. We further found that MAb S139/1 showed neutralization and hemagglutination-inhibition activities against particular strains of H1, H2, H3, and H13 subtypes of influenza A viruses. Mutant viruses that escaped neutralization by MAb S139/1 were selected from the A/Aichi/2/68 (H3N2), A/Adachi/2/57 (H2N2), and A/WSN/33 (H1N1) strains, and sequence analysis of the HA genes of these escape mutants revealed amino acid substitutions at positions 156, 158, and 193 (H3 numbering). A molecular modeling study showed that these amino acids were located on the globular head of the HA and formed a novel conformational epitope adjacent to the receptor-binding domain of HA. Furthermore, passive immunization of mice with MAb S139/1 provided heterosubtypic protection. These results demonstrate that MAb S139/1 binds to a common antigenic site shared among a variety of HA subtypes and neutralizes viral infectivity in vitro and in vivo by affecting viral attachment to cells. The present study supports the notion that cross-reactive antibodies play some roles in heterosubtypic immunity against influenza A virus infection, and underscores the potential therapeutic utility of cross-reactive antibodies against influenza. PMID- 19300498 TI - Studies of a ring-cleaving dioxygenase illuminate the role of cholesterol metabolism in the pathogenesis of Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis, the etiological agent of TB, possesses a cholesterol catabolic pathway implicated in pathogenesis. This pathway includes an iron dependent extradiol dioxygenase, HsaC, that cleaves catechols. Immuno-compromised mice infected with a DeltahsaC mutant of M. tuberculosis H37Rv survived 50% longer than mice infected with the wild-type strain. In guinea pigs, the mutant disseminated more slowly to the spleen, persisted less successfully in the lung, and caused little pathology. These data establish that, while cholesterol metabolism by M. tuberculosis appears to be most important during the chronic stage of infection, it begins much earlier and may contribute to the pathogen's dissemination within the host. Purified HsaC efficiently cleaved the catecholic cholesterol metabolite, DHSA (3,4-dihydroxy-9,10-seconandrost-1,3,5(10)-triene 9,17-dione; k(cat)/K(m) = 14.4+/-0.5 microM(-1) s(-1)), and was inactivated by a halogenated substrate analogue (partition coefficient<50). Remarkably, cholesterol caused loss of viability in the DeltahsaC mutant, consistent with catechol toxicity. Structures of HsaC:DHSA binary complexes at 2.1 A revealed two catechol-binding modes: bidentate binding to the active site iron, as has been reported in similar enzymes, and, unexpectedly, monodentate binding. The position of the bicyclo-alkanone moiety of DHSA was very similar in the two binding modes, suggesting that this interaction is a determinant in the initial substrate binding event. These data provide insights into the binding of catechols by extradiol dioxygenases and facilitate inhibitor design. PMID- 19300499 TI - A genome-wide association study confirms VKORC1, CYP2C9, and CYP4F2 as principal genetic determinants of warfarin dose. AB - We report the first genome-wide association study (GWAS) whose sample size (1,053 Swedish subjects) is sufficiently powered to detect genome-wide significance (p<1.5 x 10(-7)) for polymorphisms that modestly alter therapeutic warfarin dose. The anticoagulant drug warfarin is widely prescribed for reducing the risk of stroke, thrombosis, pulmonary embolism, and coronary malfunction. However, Caucasians vary widely (20-fold) in the dose needed for therapeutic anticoagulation, and hence prescribed doses may be too low (risking serious illness) or too high (risking severe bleeding). Prior work established that approximately 30% of the dose variance is explained by single nucleotide polymorphisms (SNPs) in the warfarin drug target VKORC1 and another approximately 12% by two non-synonymous SNPs (*2, *3) in the cytochrome P450 warfarin metabolizing gene CYP2C9. We initially tested each of 325,997 GWAS SNPs for association with warfarin dose by univariate regression and found the strongest statistical signals (p<10(-78)) at SNPs clustering near VKORC1 and the second lowest p-values (p<10(-31)) emanating from CYP2C9. No other SNPs approached genome-wide significance. To enhance detection of weaker effects, we conducted multiple regression adjusting for known influences on warfarin dose (VKORC1, CYP2C9, age, gender) and identified a single SNP (rs2108622) with genome-wide significance (p = 8.3 x 10(-10)) that alters protein coding of the CYP4F2 gene. We confirmed this result in 588 additional Swedish patients (p<0.0029) and, during our investigation, a second group provided independent confirmation from a scan of warfarin-metabolizing genes. We also thoroughly investigated copy number variations, haplotypes, and imputed SNPs, but found no additional highly significant warfarin associations. We present power analysis of our GWAS that is generalizable to other studies, and conclude we had 80% power to detect genome wide significance for common causative variants or markers explaining at least 1.5% of dose variance. These GWAS results provide further impetus for conducting large-scale trials assessing patient benefit from genotype-based forecasting of warfarin dose. PMID- 19300500 TI - A genome-wide association study of pulmonary function measures in the Framingham Heart Study. AB - The ratio of forced expiratory volume in one second to forced vital capacity (FEV(1)/FVC) is a measure used to diagnose airflow obstruction and is highly heritable. We performed a genome-wide association study in 7,691 Framingham Heart Study participants to identify single-nucleotide polymorphisms (SNPs) associated with the FEV(1)/FVC ratio, analyzed as a percent of the predicted value. Identified SNPs were examined in an independent set of 835 Family Heart Study participants enriched for airflow obstruction. Four SNPs in tight linkage disequilibrium on chromosome 4q31 were associated with the percent predicted FEV(1)/FVC ratio with p-values of genome-wide significance in the Framingham sample (best p-value = 3.6e-09). One of the four chromosome 4q31 SNPs (rs13147758; p-value 2.3e-08 in Framingham) was genotyped in the Family Heart Study and produced evidence of association with the same phenotype, percent predicted FEV(1)/FVC (p-value = 2.0e-04). The effect estimates for association in the Framingham and Family Heart studies were in the same direction, with the minor allele (G) associated with higher FEV(1)/FVC ratio levels. Results from the Family Heart Study demonstrated that the association extended to FEV(1) and dichotomous airflow obstruction phenotypes, particularly among smokers. The SNP rs13147758 was associated with the percent predicted FEV(1)/FVC ratio in independent samples from the Framingham and Family Heart Studies producing a combined p-value of 8.3e-11, and this region of chromosome 4 around 145.68 megabases was associated with COPD in three additional populations reported in the accompanying manuscript. The associated SNPs do not lie within a gene transcript but are near the hedgehog-interacting protein (HHIP) gene and several expressed sequence tags cloned from fetal lung. Though it is unclear what gene or regulatory effect explains the association, the region warrants further investigation. PMID- 19300501 TI - Genetic drift of HIV populations in culture. AB - Populations of Human Immunodeficiency Virus type 1 (HIV-1) undergo a surprisingly large amount of genetic drift in infected patients despite very large population sizes, which are predicted to be mostly deterministic. Several models have been proposed to explain this phenomenon, but all of them implicitly assume that the process of virus replication itself does not contribute to genetic drift. We developed an assay to measure the amount of genetic drift for HIV populations replicating in cell culture. The assay relies on creation of HIV populations of known size and measurements of variation in frequency of a neutral allele. Using this assay, we show that HIV undergoes approximately ten times more genetic drift than would be expected from its population size, which we defined as the number of infected cells in the culture. We showed that a large portion of the increase in genetic drift is due to non-synchronous infection of target cells. When infections are synchronized, genetic drift for the virus is only 3-fold higher than expected from its population size. Thus, the stochastic nature of biological processes involved in viral replication contributes to increased genetic drift in HIV populations. We propose that appreciation of these effects will allow better understanding of the evolutionary forces acting on HIV in infected patients. PMID- 19300502 TI - A plant germline-specific integrator of sperm specification and cell cycle progression. AB - The unique double fertilisation mechanism in flowering plants depends upon a pair of functional sperm cells. During male gametogenesis, each haploid microspore undergoes an asymmetric division to produce a large, non-germline vegetative cell and a single germ cell that divides once to produce the sperm cell pair. Despite the importance of sperm cells in plant reproduction, relatively little is known about the molecular mechanisms controlling germ cell proliferation and specification. Here, we investigate the role of the Arabidopsis male germline specific Myb protein DUO POLLEN1, DUO1, as a positive regulator of male germline development. We show that DUO1 is required for correct male germ cell differentiation including the expression of key genes required for fertilisation. DUO1 is also necessary for male germ cell division, and we show that DUO1 is required for the germline expression of the G2/M regulator AtCycB1;1 and that AtCycB1:1 can partially rescue defective germ cell division in duo1. We further show that the male germline-restricted expression of DUO1 depends upon positive promoter elements and not upon a proposed repressor binding site. Thus, DUO1 is a key regulator in the production of functional sperm cells in flowering plants that has a novel integrative role linking gametic cell specification and cell cycle progression. PMID- 19300503 TI - Capturing the spectrum of interaction effects in genetic association studies by simulated evaporative cooling network analysis. AB - Evidence from human genetic studies of several disorders suggests that interactions between alleles at multiple genes play an important role in influencing phenotypic expression. Analytical methods for identifying Mendelian disease genes are not appropriate when applied to common multigenic diseases, because such methods investigate association with the phenotype only one genetic locus at a time. New strategies are needed that can capture the spectrum of genetic effects, from Mendelian to multifactorial epistasis. Random Forests (RF) and Relief-F are two powerful machine-learning methods that have been studied as filters for genetic case-control data due to their ability to account for the context of alleles at multiple genes when scoring the relevance of individual genetic variants to the phenotype. However, when variants interact strongly, the independence assumption of RF in the tree node-splitting criterion leads to diminished importance scores for relevant variants. Relief-F, on the other hand, was designed to detect strong interactions but is sensitive to large backgrounds of variants that are irrelevant to classification of the phenotype, which is an acute problem in genome-wide association studies. To overcome the weaknesses of these data mining approaches, we develop Evaporative Cooling (EC) feature selection, a flexible machine learning method that can integrate multiple importance scores while removing irrelevant genetic variants. To characterize detailed interactions, we construct a genetic-association interaction network (GAIN), whose edges quantify the synergy between variants with respect to the phenotype. We use simulation analysis to show that EC is able to identify a wide range of interaction effects in genetic association data. We apply the EC filter to a smallpox vaccine cohort study of single nucleotide polymorphisms (SNPs) and infer a GAIN for a collection of SNPs associated with adverse events. Our results suggest an important role for hubs in SNP disease susceptibility networks. The software is available at (http://sites.google.com/site/McKinneyLab/software). PMID- 19300504 TI - Positional information generated by spatially distributed signaling cascades. AB - The temporal and stationary behavior of protein modification cascades has been extensively studied, yet little is known about the spatial aspects of signal propagation. We have previously shown that the spatial separation of opposing enzymes, such as a kinase and a phosphatase, creates signaling activity gradients. Here we show under what conditions signals stall in the space or robustly propagate through spatially distributed signaling cascades. Robust signal propagation results in activity gradients with long plateaus, which abruptly decay at successive spatial locations. We derive an approximate analytical solution that relates the maximal amplitude and propagation length of each activation profile with the cascade level, protein diffusivity, and the ratio of the opposing enzyme activities. The control of the spatial signal propagation appears to be very different from the control of transient temporal responses for spatially homogenous cascades. For spatially distributed cascades where activating and deactivating enzymes operate far from saturation, the ratio of the opposing enzyme activities is shown to be a key parameter controlling signal propagation. The signaling gradients characteristic for robust signal propagation exemplify a pattern formation mechanism that generates precise spatial guidance for multiple cellular processes and conveys information about the cell size to the nucleus. PMID- 19300505 TI - Robust food anticipatory activity in BMAL1-deficient mice. AB - Food availability is a potent environmental cue that directs circadian locomotor activity in rodents. Even though nocturnal rodents prefer to forage at night, daytime food anticipatory activity (FAA) is observed prior to short meals presented at a scheduled time of day. Under this restricted feeding regimen, rodents exhibit two distinct bouts of activity, a nocturnal activity rhythm that is entrained to the light-dark cycle and controlled by the master clock in the suprachiasmatic nuclei (SCN) and a daytime bout of activity that is phase-locked to mealtime. FAA also occurs during food deprivation, suggesting that a food entrainable oscillator (FEO) keeps time in the absence of scheduled feeding. Previous studies have demonstrated that the FEO is anatomically distinct from the SCN and that FAA is observed in mice lacking some circadian genes essential for timekeeping in the SCN. In the current study, we optimized the conditions for examining FAA during restricted feeding and food deprivation in mice lacking functional BMAL1, which is critical for circadian rhythm generation in the SCN. We found that BMAL1-deficient mice displayed FAA during restricted feeding in 12hr light:12hr dark (12L:12D) and 18L:6D lighting cycles, but distinct activity during food deprivation was observed only in 18L:6D. While BMAL1-deficient mice also exhibited robust FAA during restricted feeding in constant darkness, mice were hyperactive during food deprivation so it was not clear that FAA consistently occurred at the time of previously scheduled food availability. Taken together, our findings suggest that optimization of experimental conditions such as photoperiod may be necessary to visualize FAA in genetically modified mice. Furthermore, the expression of FAA may be possible without a circadian oscillator that depends on BMAL1. PMID- 19300506 TI - PP1gamma2 and PPP1R11 are parts of a multimeric complex in developing testicular germ cells in which their steady state levels are reciprocally related. AB - Mice lacking the protein phosphatase 1 gamma isoforms, PP1gamma1 and PP1gamma2, are male-sterile due to defective germ cell morphogenesis and apoptosis. However, this deficiency causes no obvious abnormality in other tissues. A biochemical approach was employed to learn how expression versus deficiency of PP1gamma2, the predominant PP1 isoform in male germ cells, affects spermatogenesis. Methods used in this study include column chromatography, western blot and northern blot analyses, GST pull-down assays, immunoprecipitation, non-denaturing gel electrophoresis, phosphatase enzyme assays, protein sequencing, and immunohistochemistry. We report for the first time that in wild-type testis, PP1gamma2 forms an inactive complex with actin, protein phosphatase 1 regulatory subunit 7 (PPP1R7), and protein phosphatase 1 regulatory subunit 11 (PPP1R11), the latter, a potent PP1 inhibitor. Interestingly, PPP1R11 protein, but not its mRNA level, falls significantly in PP1gamma-null testis where mature sperm are virtually absent. Conversely, both mature sperm numbers and the PPP1R11 level increase substantially in PP1gamma-null testis expressing transgenic PP1gamma2. PPP1R11 also appears to be ubiquitinated in PP1gamma-null testis. The levels of PP1gamma2 and PPP1R11 were increased in phenotypically normal PP1alpha-null testis. However, in PP1alpha-null spleen, where PP1gamma2 normally is not expressed, PPP1R11 levels remained unchanged. Our data clearly show a direct reciprocal relationship between the levels of the protein phosphatase isoform PP1gamma2 and its regulator PPP1R11, and suggest that complex formation between these polypeptides in testis may prevent proteolysis of PPP1R11 and thus, germ cell apoptosis. PMID- 19300507 TI - The generation of forces and moments during visual-evoked steering maneuvers in flying Drosophila. AB - Sideslip force, longitudinal force, rolling moment, and pitching moment generated by tethered fruit flies, Drosophila melanogaster, were measured during optomotor reactions within an electronic flight simulator. Forces and torques were acquired by optically measuring the angular deflections of the beam to which the flies were tethered using a laser and a photodiode. Our results indicate that fruit flies actively generate both sideslip and roll in response to a lateral focus of expansion (FOE). The polarity of this behavior was such that the animal's aerodynamic response would carry it away from the expanding pattern, suggesting that it constitutes an avoidance reflex or centering response. Sideslip forces and rolling moments were sinusoidal functions of FOE position, whereas longitudinal force was proportional to the absolute value of the sine of FOE position. Pitching moments remained nearly constant irrespective of stimulus position or strength, with a direction indicating a tonic nose-down pitch under tethered conditions. These experiments expand our understanding of the degrees of freedom that a fruit fly can actually control in flight. PMID- 19300508 TI - Oral probiotic control skin inflammation by acting on both effector and regulatory T cells. AB - Probiotics are believed to alleviate allergic and inflammatory skin disorders, but their impact on pathogenic effector T cells remains poorly documented. Here we show that oral treatment with the probiotic bacteria L. casei (DN-114 001) alone alleviates antigen-specific skin inflammation mediated by either protein specific CD4(+) T cells or hapten-specific CD8(+) T cells. In the model of CD8(+) T cell-mediated skin inflammation, which reproduces allergic contact dermatitis in human, inhibition of skin inflammation by L. casei is not due to impaired priming of hapten-specific IFNgamma-producing cytolytic CD8(+) effector T cells. Alternatively, L. casei treatment reduces the recruitment of CD8(+) effector T cells into the skin during the elicitation (i.e. symptomatic) phase of CHS. Inhibition of skin inflammation by L. casei requires MHC class II-restricted CD4(+) T cells but not CD1d-restricted NK-T cells. L casei treatment enhanced the frequency of FoxP3(+) Treg in the skin and increased the production of IL-10 by CD4(+)CD25(+) regulatory T cells in skin draining lymph nodes of hapten sensitized mice. These data demonstrate that orally administered L. casei (DN-114 001) efficiently alleviate T cell-mediated skin inflammation without causing immune suppression, via mechanisms that include control of CD8(+) effector T cells and involve regulatory CD4(+) T cells. L. casei (DN-114 001) may thus represent a probiotic of potential interest for immunomodulation of T cell mediated allergic skin diseases in human. PMID- 19300509 TI - Emergent genome-wide control in wildtype and genetically mutated lipopolysaccarides-stimulated macrophages. AB - Large-scale gene expression studies have mainly focused on highly expressed and 'discriminatory' genes to decipher key regulatory processes. Biological responses are consequence of the concerted action of gene regulatory network, thus, limiting our attention to genes having the most significant variations is insufficient for a thorough understanding of emergent whole genome response. Here we comprehensively analyzed the temporal oligonucleotide microarray data of lipopolysaccharide (LPS) stimulated macrophages in 4 genotypes; wildtype, Myeloid Differentiation factor 88 (MyD88) knockout (KO), TIR-domain-containing adapter inducing interferon-beta (TRIF) KO and MyD88/TRIF double KO (DKO). Pearson correlations computed on the whole genome expression between different genotypes are extremely high (>0.98), indicating a strong co-regulation of the entire expression network. Further correlation analyses reveal genome-wide response is biphasic, i) acute-stochastic mode consisting of small number of sharply induced immune-related genes and ii) collective mode consisting of majority of weakly induced genes of diverse cellular processes which collectively adjust their expression level. Notably, temporal correlations of a small number of randomly selected genes from collective mode show scalability. Furthermore, in collective mode, the transition from large scatter in expression distributions for single ORFs to smooth linear lines emerges as an organizing principle when grouping of 50 ORFs and above. With this emergent behavior, the role of MyD88, TRIF and novel MyD88, TRIF-independent processes for gene induction can be linearly superposed to decipher quantitative whole genome differential control of transcriptional and mRNA decay machineries. Our work demonstrates genome-wide co-regulated responses subsequent to specific innate immune stimulus which have been largely neglected. PMID- 19300510 TI - The DISC1 pathway modulates expression of neurodevelopmental, synaptogenic and sensory perception genes. AB - BACKGROUND: Genetic and biological evidence supports a role for DISC1 across a spectrum of major mental illnesses, including schizophrenia and bipolar disorder. There is evidence for genetic interplay between variants in DISC1 and in biologically interacting loci in psychiatric illness. DISC1 also associates with normal variance in behavioral and brain imaging phenotypes. METHODOLOGY: Here, we analyze public domain datasets and demonstrate correlations between variants in the DISC1 pathway genes and levels of gene expression. Genetic variants of DISC1, NDE1, PDE4B and PDE4D regulate the expression of cytoskeletal, synaptogenic, neurodevelopmental and sensory perception proteins. Interestingly, these regulated genes include existing targets for drug development in depression and psychosis. CONCLUSIONS: Our systematic analysis provides further evidence for the relevance of the DISC1 pathway to major mental illness, identifies additional potential targets for therapeutic intervention and establishes a general strategy to mine public datasets for insights into disease pathways. PMID- 19300511 TI - Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. AB - We describe the small-scale, laboratory-based, production and quality control of a cocktail, consisting of exclusively lytic bacteriophages, designed for the treatment of Pseudomonas aeruginosa and Staphylococcus aureus infections in burn wound patients. Based on successive selection rounds three bacteriophages were retained from an initial pool of 82 P. aeruginosa and 8 S. aureus bacteriophages, specific for prevalent P. aeruginosa and S. aureus strains in the Burn Centre of the Queen Astrid Military Hospital in Brussels, Belgium. This cocktail, consisting of P. aeruginosa phages 14/1 (Myoviridae) and PNM (Podoviridae) and S. aureus phage ISP (Myoviridae) was produced and purified of endotoxin. Quality control included Stability (shelf life), determination of pyrogenicity, sterility and cytotoxicity, confirmation of the absence of temperate bacteriophages and transmission electron microscopy-based confirmation of the presence of the expected virion morphologic particles as well as of their specific interaction with the target bacteria. Bacteriophage genome and proteome analysis confirmed the lytic nature of the bacteriophages, the absence of toxin-coding genes and showed that the selected phages 14/1, PNM and ISP are close relatives of respectively F8, phiKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee. PMID- 19300512 TI - The role of genomic data in the discovery, annotation and evolutionary interpretation of the interferon-lambda family. AB - BACKGROUND: Type-I interferons, type-II interferons, and the IL-10 family are helical cytokines with similar three-dimensional folds. However, their homologous relationship is difficult to detect on the basis of sequence alone. We have previously described the discovery of the human type-III interferons (IFN lambda 1, -2, -3 or IL-29, IL-28A, IL-28B), which required a combination of manual and computational techniques applied to predicted protein sequences. PRINCIPAL FINDINGS: Here we describe how the use of gene structure analysis and comparative genomics enabled a more extensive understanding of these genes early in the discovery process. More recently, additional mammalian genome sequences have shown that there are between one and potentially nine copies of interferon lambda genes in each genome, and that several species have single exon versions of the interferon lambda gene. SIGNIFICANCE: The variable number of single exon type-I interferons in mammals, along with recently identified genes in zebrafish homologous to interferons allows a story of interferon evolution to be proposed. This model suggests that the gene duplications and single exon retrotransposons of mammalian type-III interferons are positively selected for within a genome. These characteristics are also shared with the fish interferons and could be responsible for the generation of the IL10 family and also the single exon type-I interferons. PMID- 19300514 TI - Producer nutritional quality controls ecosystem trophic structure. AB - Trophic structure, or the distribution of biomass among producers and consumers, determines key ecosystem values, such as the abundance of infectious, harvestable or conservation target species, and the storage and cycling of carbon and nutrients. There has been much debate on what controls ecosystem trophic structure, yet the answer is still elusive. Here we show that the nutritional quality of primary producers controls the trophic structure of ecosystems. By increasing the efficiency of trophic transfer, higher producer nutritional quality results in steeper ecosystem trophic structure, and those changes are more pronounced in terrestrial than in aquatic ecosystems probably due to the more stringent nutritional limitation of terrestrial herbivores. These results explain why ecosystems composed of highly nutritional primary producers feature high consumer productivity, fast energy recycling, and reduced carbon accumulation. Anthropogenic changes in producer nutritional quality, via changes in trophic structure, may alter the values and functions of ecosystems, and those alterations may be more important in terrestrial ecosystems. PMID- 19300513 TI - Matrix metalloproteinase proteolysis of the myelin basic protein isoforms is a source of immunogenic peptides in autoimmune multiple sclerosis. AB - BACKGROUND: Matrix metalloproteinases (MMPs) play a significant role in the fragmentation of myelin basic protein (MBP) and demyelination leading to autoimmune multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). The classic MBP isoforms are predominantly expressed in the oligodendrocytes of the CNS. The splice variants of the single MBP gene (Golli MBP BG21 and J37) are widely expressed in the neurons and also in the immune cells. The relative contribution of the individual MMPs to the MBP cleavage is not known. METHODOLOGY/PRINCIPAL FINDINGS: To elucidate which MMP plays the primary role in cleaving MBP, we determined the efficiency of MMP-2, MMP-8, MMP 9, MMP-10, MMP-12, MT1-MMP, MT2-MMP, MT3-MMP, MT4-MMP, MT5-MMP and MT6-MMP in the cleavage of the MBP, BG21 and J37 isoforms in the in vitro cleavage reactions followed by mass-spectroscopy analysis of the cleavage fragments. As a result, we identified the MMP cleavage sites and the sequence of the resulting fragments. We determined that MBP, BG21 and J37 are highly sensitive to redundant MMP proteolysis. MT6-MMP (initially called leukolysin), however, was superior over all of the other MMPs in cleaving the MBP isoforms. Using the mixed lymphocyte culture assay, we demonstrated that MT6-MMP proteolysis of the MBP isoforms readily generated, with a near quantitative yield, the immunogenic N-terminal 1 15 MBP peptide. This peptide selectively stimulated the proliferation of the PGPR7.5 T cell clone isolated from mice with EAE and specific for the 1-15 MBP fragment presented in the MHC H-2(U) context. CONCLUSIONS/SIGNIFICANCE: In sum, our biochemical observations led us to hypothesize that MT6-MMP, which is activated by furin and associated with the lipid rafts, plays an important role in MS pathology and that MT6-MMP is a novel and promising drug target in MS especially when compared with other individual MMPs. PMID- 19300515 TI - Directing experimental biology: a case study in mitochondrial biogenesis. AB - Computational approaches have promised to organize collections of functional genomics data into testable predictions of gene and protein involvement in biological processes and pathways. However, few such predictions have been experimentally validated on a large scale, leaving many bioinformatic methods unproven and underutilized in the biology community. Further, it remains unclear what biological concerns should be taken into account when using computational methods to drive real-world experimental efforts. To investigate these concerns and to establish the utility of computational predictions of gene function, we experimentally tested hundreds of predictions generated from an ensemble of three complementary methods for the process of mitochondrial organization and biogenesis in Saccharomyces cerevisiae. The biological data with respect to the mitochondria are presented in a companion manuscript published in PLoS Genetics (doi:10.1371/journal.pgen.1000407). Here we analyze and explore the results of this study that are broadly applicable for computationalists applying gene function prediction techniques, including a new experimental comparison with 48 genes representing the genomic background. Our study leads to several conclusions that are important to consider when driving laboratory investigations using computational prediction approaches. While most genes in yeast are already known to participate in at least one biological process, we confirm that genes with known functions can still be strong candidates for annotation of additional gene functions. We find that different analysis techniques and different underlying data can both greatly affect the types of functional predictions produced by computational methods. This diversity allows an ensemble of techniques to substantially broaden the biological scope and breadth of predictions. We also find that performing prediction and validation steps iteratively allows us to more completely characterize a biological area of interest. While this study focused on a specific functional area in yeast, many of these observations may be useful in the contexts of other processes and organisms. PMID- 19300516 TI - Bim and Bmf synergize to induce apoptosis in Neisseria gonorrhoeae infection. AB - Bcl-2 family proteins including the pro-apoptotic BH3-only proteins are central regulators of apoptotic cell death. Here we show by a focused siRNA miniscreen that the synergistic action of the BH3-only proteins Bim and Bmf is required for apoptosis induced by infection with Neisseria gonorrhoeae (Ngo). While Bim and Bmf were associated with the cytoskeleton of healthy cells, they both were released upon Ngo infection. Loss of Bim and Bmf from the cytoskeleton fraction required the activation of Jun-N-terminal kinase-1 (JNK-1), which in turn depended on Rac-1. Depletion and inhibition of Rac-1, JNK-1, Bim, or Bmf prevented the activation of Bak and Bax and the subsequent activation of caspases. Apoptosis could be reconstituted in Bim-depleted and Bmf-depleted cells by additional silencing of antiapoptotic Mcl-1 and Bcl-X(L), respectively. Our data indicate a synergistic role for both cytoskeletal-associated BH3-only proteins, Bim, and Bmf, in an apoptotic pathway leading to the clearance of Ngo infected cells. PMID- 19300517 TI - Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. AB - Rapid antibiotic susceptibility testing of Mycobacterium tuberculosis is of paramount importance as multiple- and extensively-drug resistant strains of M. tuberculosis emerge and spread. We describe here a virus-based assay in which fluoromycobacteriophages are used to deliver a GFP or ZsYellow fluorescent marker gene to M. tuberculosis, which can then be monitored by fluorescent detection approaches including fluorescent microscopy and flow cytometry. Pre-clinical evaluations show that addition of either Rifampicin or Streptomycin at the time of phage addition obliterates fluorescence in susceptible cells but not in isogenic resistant bacteria enabling drug sensitivity determination in less than 24 hours. Detection requires no substrate addition, fewer than 100 cells can be identified, and resistant bacteria can be detected within mixed populations. Fluorescence withstands fixation by paraformaldehyde providing enhanced biosafety for testing MDR-TB and XDR-TB infections. PMID- 19300518 TI - De-novo identification of PPARgamma/RXR binding sites and direct targets during adipogenesis. AB - BACKGROUND: The pathophysiology of obesity and type 2 diabetes mellitus is associated with abnormalities in endocrine signaling in adipose tissue and one of the key signaling affectors operative in these disorders is the nuclear hormone transcription factor peroxisome proliferator-activated receptor-gamma (PPARgamma). PPARgamma has pleiotropic functions affecting a wide range of fundamental biological processes including the regulation of genes that modulate insulin sensitivity, adipocyte differentiation, inflammation and atherosclerosis. To date, only a limited number of direct targets for PPARgamma have been identified through research using the well established pre-adipogenic cell line, 3T3-L1. In order to obtain a genome-wide view of PPARgamma binding sites, we applied the pair end-tagging technology (ChIP-PET) to map PPARgamma binding sites in 3T3-L1 preadipocyte cells. METHODOLOGY/PRINCIPAL FINDINGS: Coupling gene expression profile analysis with ChIP-PET, we identified in a genome-wide manner over 7700 DNA binding sites of the transcription factor PPARgamma and its heterodimeric partner RXR during the course of adipocyte differentiation. Our validation studies prove that the identified sites are bona fide binding sites for both PPARgamma and RXR and that they are functionally capable of driving PPARgamma specific transcription. Our results strongly indicate that PPARgamma is the predominant heterodimerization partner for RXR during late stages of adipocyte differentiation. Additionally, we find that PPARgamma/RXR association is enriched within the proximity of the 5' region of the transcription start site and this association is significantly associated with transcriptional up regulation of genes involved in fatty acid and lipid metabolism confirming the role of PPARgamma as the master transcriptional regulator of adipogenesis. Evolutionary conservation analysis of these binding sites is greater when adjacent to up-regulated genes than down-regulated genes, suggesting the primordial function of PPARgamma/RXR is in the induction of genes. Our functional validations resulted in identifying novel PPARgamma direct targets that have not been previously reported to promote adipogenic differentiation. CONCLUSIONS/SIGNIFICANCE: We have identified in a genome-wide manner the binding sites of PPARgamma and RXR during the course of adipogenic differentiation in 3T3L1 cells, and provide an important resource for the study of PPARgamma function in the context of adipocyte differentiation. PMID- 19300519 TI - Cationic polybutyl cyanoacrylate nanoparticles for DNA delivery. AB - To enhance the intracellular delivery potential of plasmid DNA using nonviral vectors, we used polybutyl cyanoacrylate (PBCA) and chitosan to prepare PBCA nanoparticles (NPs) by emulsion polymerization and prepared NP/DNA complexes through the complex coacervation of nanoparticles with the DNA. The object of our work is to evaluate the characterization and transfection efficiency of PBCA-NPs. The NPs have a zeta potential of 25.53 mV at pH 7.4 and size about 200 nm. Electrophoretic analysis suggested that the NPs with positive charges could protect the DNA from nuclease degradation and cell viability assay showed that the NPs exhibit a low cytotoxicity to human hepatocellular carcinoma (HepG2) cells. Qualitative and quantitative analysis of transfection in HepG2 cells by the nanoparticles carrying plasmid DNA encoding for enhanced green fluorescent protein (EGFP-N1) was done by digital fluorescence imaging microscopy system and fluorescence-activated cell sorting (FACS). Qualitative results showed highly efficient expression of GFP that remained stable for up to 96 hours. Quantitative results from FACS showed that PBCA-NPs were significantly more effective in transfecting HepG2 cells after 72 hours postincubation. The results of this study suggested that PBCA-NPs have favorable properties for nonviral delivery. PMID- 19300520 TI - Anxiolytic and anticonvulsant effects on mice of flavonoids, linalool, and alpha tocopherol presents in the extract of leaves of Cissus sicyoides L. (Vitaceae). AB - The aim of the present study is to demonstrate the anxiolytic and anticonvulsant effects of a hydroalcoholic extract obtained from the aerial parts of Cissus sicyoides L. (CS) (Vitaceae) on male and female mice using several behavioral assays. Groups of males and females treated via intraperitoneal (IP) with doses of 300, 600, and 1000 mg/kg of the extract showed significant action in the elevated plus-maze (EPM), time spent in the open arms, and number of entries in the open arms. The board-hole test also showed a significant increase in the time spent in head-dipping and in marble-burying test of the number of marbles buried. The same treatment increased the duration of sleeping time induced by sodium pentobarbital and also showed a significant increase in protection against pentylenotetrazole-induced convulsions. These results indicate an anxiolytic and anticonvulsant-like action from C. sicyoides L. extract on mice, probably due to the action of flavonoid(s), Linalool, and alpha-tocopherol present in the C. sicyoides leaves. PMID- 19300521 TI - Serum insulin-like growth factor-1 and nitric oxide levels in Parkinson's disease. AB - The aim of this study was to investigate the role of circulating growth hormone (GH), insulin growth factor-1 (IGF-1), IGF binding protein-3 (IGFBP-3), and nitric oxide (NO) concentrations in the patients suffering from Parkinson's disease (PD). The study groups were consisted of 25 patients with PD and 25 matched healthy subjects as a control. The NO level of patients in PD group (2.3 +/- 0.4 micromol/L) was significantly lower than that in the control group (2.8 +/- 0.6 micromol/L) (P:.011). Although there were no statistically significant differences in the GH, IGF-1, and IGF BP-3 levels among the two groups, in this preliminary study, we found low NO and mildly elevated IGF-1 levels in the patients with PD. The results may be associated with adaptation or protective mechanisms in the neurodegenerative disease processes such as seen in the PD. Further studies should be carried out to confirm our results. PMID- 19300522 TI - Sparse representation for classification of tumors using gene expression data. AB - Personalized drug design requires the classification of cancer patients as accurate as possible. With advances in genome sequencing and microarray technology, a large amount of gene expression data has been and will continuously be produced from various cancerous patients. Such cancer-alerted gene expression data allows us to classify tumors at the genomewide level. However, cancer alerted gene expression datasets typically have much more number of genes (features) than that of samples (patients), which imposes a challenge for classification of tumors. In this paper, a new method is proposed for cancer diagnosis using gene expression data by casting the classification problem as finding sparse representations of test samples with respect to training samples. The sparse representation is computed by the l(1)-regularized least square method. To investigate its performance, the proposed method is applied to six tumor gene expression datasets and compared with various support vector machine (SVM) methods. The experimental results have shown that the performance of the proposed method is comparable with or better than those of SVMs. In addition, the proposed method is more efficient than SVMs as it has no need of model selection. PMID- 19300523 TI - Identification of IbeR as a stationary-phase regulator in meningitic Escherichia coli K1 that carries a loss-of-function mutation in rpoS. AB - IbeR is a regulator present in meningitic Escherichia coli strain E44 that carries a loss-of-function mutation in the stationary-phase (SP) regulatory gene rpoS. In order to determine whether IbeR is an SP regulator in E44, two dimensional gel electrophoresis and LC-MS were used to compare the proteomes of a noninvasive ibeR deletion mutant BR2 and its parent strain E44 in the SP. Four up regulated (TufB, GapA, OmpA, AhpC) and three down-regulated (LpdA, TnaA, OpmC) proteins in BR2 were identified when compared to E44. All these proteins contribute to energy metabolism or stress resistance, which is related to SP regulation. One of the down-regulated proteins, tryptophanase (TnaA), which is regulated by RpoS in other E. coli strains, is associated with SP regulation via production of a signal molecule indole. Our studies demonstrated that TnaA was required for E44 invasion, and that indole was able to restore the noninvasive phenotype of the tnaA mutant. The production of indole was significantly reduced in BR2, indicating that ibeR is required for the indole production via tnaA. Survival studies under different stress conditions indicated that IbeR contributed to bacteria stress resistance in the SP. Taken together, IbeR is a novel regulator contributing to the SP regulation. PMID- 19300524 TI - Molecularly characterised xenograft tumour mouse models: valuable tools for evaluation of new therapeutic strategies for secondary liver cancers. AB - To develop and evaluate new therapeutic strategies for the treatment of human cancers, well-characterised preclinical model systems are a prerequisite. To this aim, we have established xenotransplantation mouse models and corresponding cell cultures from surgically obtained secondary human liver tumours. Established xenograft tumours were patho- and immunohistologically characterised, and expression levels of cancer-relevant genes were quantified in paired original and xenograft tumours and the derivative cell cultures applying RT-PCR-based array technology. Most of the characteristic morphological and immunohistochemical features of the original tumours were shown to be maintained. No differences were found concerning expression of genes involved in cell cycle regulation and oncogenesis. Interestingly, cytokine and matrix metalloproteinase encoding genes appeared to be expressed differentially. Thus, the established models are closely reflecting pathohistological and molecular characteristics of the selected human tumours and may therefore provide useful tools for preclinical analyses of new antitumour strategies in vivo. PMID- 19300525 TI - Plant genomics. PMID- 19300526 TI - Decreased ADP-ribosyl cyclase activity in peripheral blood mononuclear cells from diabetic patients with nephropathy. AB - AIMS/HYPOTHESIS: ADP-ribosyl-cyclase activity (ADPRCA) of CD38 and other ectoenzymes mainly generate cyclic adenosine 5'diphosphate-(ADP-) ribose (cADPR) as a second messenger in various mammalian cells, including pancreatic beta cells and peripheral blood mononuclear cells (PBMCs). Since PBMCs contribute to the pathogenesis of diabetic nephropathy, ADPRCA of PBMCs could serve as a clinical prognostic marker for diabetic nephropathy. This study aimed to investigate the connection between ADPRCA in PBMCs and diabetic complications. METHODS: PBMCs from 60 diabetic patients (10 for type 1 and 50 for type 2) and 15 nondiabetic controls were fluorometrically measured for ADPRCA based on the conversion of nicotinamide guanine dinucleotide (NGD(+)) into cyclic GDP-ribose. RESULTS: ADPRCA negatively correlated with the level of HbA1c (P = .040, R(2) = .073), although ADPRCA showed no significant correlation with gender, age, BMI, blood pressure, level of fasting plasma glucose and lipid levels, as well as type, duration, or medication of diabetes. Interestingly, patients with nephropathy, but not other complications, presented significantly lower ADPRCA than those without nephropathy (P = .0198) and diabetes (P = .0332). ANCOVA analysis adjusted for HbA1c showed no significant correlation between ADPRCA and nephropathy. However, logistic regression analyses revealed that determinants for nephropathy were systolic blood pressure and ADPRCA, not HbA1c. CONCLUSION/INTERPRETATION: Decreased ADPRCA significantly correlated with diabetic nephropathy. ADPRCA in PBMCs would be an important marker associated with diabetic nephropathy. PMID- 19300527 TI - Reconstructing generalized logical networks of transcriptional regulation in mouse brain from temporal gene expression data. AB - Gene expression time course data can be used not only to detect differentially expressed genes but also to find temporal associations among genes. The problem of reconstructing generalized logical networks to account for temporal dependencies among genes and environmental stimuli from transcriptomic data is addressed. A network reconstruction algorithm was developed that uses statistical significance as a criterion for network selection to avoid false-positive interactions arising from pure chance. The multinomial hypothesis testing-based network reconstruction allows for explicit specification of the false-positive rate, unique from all extant network inference algorithms. The method is superior to dynamic Bayesian network modeling in a simulation study. Temporal gene expression data from the brains of alcohol-treated mice in an analysis of the molecular response to alcohol are used for modeling. Genes from major neuronal pathways are identified as putative components of the alcohol response mechanism. Nine of these genes have associations with alcohol reported in literature. Several other potentially relevant genes, compatible with independent results from literature mining, may play a role in the response to alcohol. Additional, previously unknown gene interactions were discovered that, subject to biological verification, may offer new clues in the search for the elusive molecular mechanisms of alcoholism. PMID- 19300528 TI - Functional classification of genome-scale metabolic networks. AB - We propose two strategies to characterize organisms with respect to their metabolic capabilities. The first, investigative, strategy describes metabolic networks in terms of their capability to utilize different carbon sources, resulting in the concept of carbon utilization spectra. In the second, predictive, approach minimal nutrient combinations are predicted from the structure of the metabolic networks, resulting in a characteristic nutrient profile. Both strategies allow for a quantification of functional properties of metabolic networks, allowing to identify groups of organisms with similar functions. We investigate whether the functional description reflects the typical environments of the corresponding organisms by dividing all species into disjoint groups based on whether they are aerotolerant and/or photosynthetic. Despite differences in the underlying concepts, both measures display some common features. Closely related organisms often display a similar functional behavior and in both cases the functional measures appear to correlate with the considered classes of environments. Carbon utilization spectra and nutrient profiles are complementary approaches toward a functional classification of organism-wide metabolic networks. Both approaches contain different information and thus yield different clusterings, which are both different from the classical taxonomy of organisms. Our results indicate that a sophisticated combination of our approaches will allow for a quantitative description reflecting the lifestyles of organisms. PMID- 19300529 TI - Effects of active site mutations in haemoglobin I from Lucina pectinata: a molecular dynamic study. AB - Haemoglobin I from Lucina pectinata is a monomeric protein consisting of 142 amino acids. Its active site contains a peculiar arrangement of phenylalanine residues (PheB10, PheCD1 and PheE11) and a distal Gln at position E7. Active site mutations at positions B10, E7 and E11 were performed in deoxy haemoglobin I (HbI), followed by 10 ns molecular dynamic simulations. The results showed that the mutations induced changes in domains far from the active site producing more flexible structures than the native HbI. Distance analyses revealed that the heme pocket amino acids at positions E7 and B10 are extremely sensitive to any heme pocket residue mutation. The high flexibility observed by the E7 position suggests an important role in the ligand binding kinetics in ferrous HbI, while both positions play a major role in the ligand stabilisation processes. Furthermore, our results showed that E11Phe plays a pivotal role in protein stability. PMID- 19300530 TI - Serum angiopoietin-1 and -2 levels discriminate cerebral malaria from uncomplicated malaria and predict clinical outcome in African children. AB - BACKGROUND: Limited tools exist to identify which individuals infected with Plasmodium falciparum are at risk of developing serious complications such as cerebral malaria (CM). The objective of this study was to assess serum biomarkers that differentiate between CM and non-CM, with the long-term goal of developing a clinically informative prognostic test for severe malaria. METHODOLOGY/PRINCIPAL FINDINGS: Based on the hypothesis that endothelial activation and blood-brain barrier dysfunction contribute to CM pathogenesis, we examined the endothelial regulators, angiopoietin-1 (ANG-1) and angiopoietin-2 (ANG-2), in serum samples from P. falciparum-infected patients with uncomplicated malaria (UM) or CM, from two diverse populations--Thai adults and Ugandan children. Angiopoietin levels were compared to tumour necrosis factor (TNF). In both populations, ANG-1 levels were significantly decreased and ANG-2 levels were significantly increased in CM versus UM and healthy controls (p<0.001). TNF was significantly elevated in CM in the Thai adult population (p<0.001), but did not discriminate well between CM and UM in African children. Receiver operating characteristic curve analysis showed that ANG-1 and the ratio of ANG-2:ANG-1 accurately discriminated CM patients from UM in both populations. Applied as a diagnostic test, ANG-1 had a sensitivity and specificity of 100% for distinguishing CM from UM in Thai adults and 70% and 75%, respectively, for Ugandan children. Across both populations the likelihood ratio of CM given a positive test (ANG-1<15 ng/mL) was 4.1 (2.7-6.5) and the likelihood ratio of CM given a negative test was 0.29 (0.20-0.42). Moreover, low ANG-1 levels at presentation predicted subsequent mortality in children with CM (p = 0.027). CONCLUSIONS/SIGNIFICANCE: ANG-1 and the ANG-2/1 ratio are promising clinically informative biomarkers for CM. Additional studies should address their utility as prognostic biomarkers and potential therapeutic targets in severe malaria. PMID- 19300531 TI - Negative affect and ruminative self-focus during everyday goal pursuit. AB - Models of self-regulation propose that negative affect is generated when progress towards goals is perceived to be inadequate. Similarly, ruminative thinking is hypothesised to be triggered by unattained goals (Martin & Tesser, 1996). We conducted an experience-sampling study in which participants recorded their negative affect, ruminative self-focus, and goal appraisals eight times daily for one week. Negative affect and ruminative self-focus were each associated with low levels of goal success and (with the exception of sadness) high levels of goal importance. As predicted, the combination of low goal success and high goal importance was associated with the highest levels of negative affect, and this interaction was marginally significant for ruminative self-focus. Decomposition of the ruminative self-focus measure revealed that the success by importance interaction was significantly associated with focus on problems but not focus on feelings. Findings did not differ for individuals reporting high versus low levels of depressive symptoms or trait rumination. These results suggest that self-regulatory models of goal pursuit provide a useful explanatory framework for the study of affect and ruminative thinking in everyday life. PMID- 19300533 TI - Assessing Sexual Attitudes and Behaviors of Young Women: A Joint Model with Nonlinear Time Effects, Time Varying Covariates, and Dropouts. AB - Understanding human sexual behaviors is essential for the effective prevention of sexually transmitted infections. Analysis of longitudinally measured sexual behavioral data, however, is often complicated by zero-inflation of event counts, nonlinear time trend, time-varying covariates, and informative dropouts. Ignoring these complicating factors could undermine the validity of the study findings. In this paper, we put forth a unified joint modeling structure that accommodates these features of the data. Specifically, we propose a pair of simultaneous models for the zero-inflated event counts: Each of these models contains an auto regressive structure for the accommodation of the effect of recent event history, and a nonparametric component for the modeling of nonlinear time effect. Informative dropout and time varying covariates are modeled explicitly in the process. Model fitting and parameter estimation are carried out in a Bayesian paradigm by the use of a Markov Chain Monte Carlo (MCMC) method. Analytical results showed that adolescent sexual behaviors tended to evolve nonlinearly over time and they were strongly influenced by the day-to-day variations in mood and sexual interests. These findings suggest that adolescent sex is to a large extent driven by intrinsic factors rather than being compelled by circumstances, thus highlighting the need of education on self protective measures against infection risks. PMID- 19300532 TI - Death in heart failure: a community perspective. AB - BACKGROUND: Mortality in heart failure (HF) remains high but causes of death are incompletely defined. As HF is heterogeneous syndrome categorized according to ejection fraction (EF), the association between EF and causes of death is important, yet elusive. METHOD AND RESULTS: Community subjects with HF were classified according to preserved (> or =50%) and reduced EF (<50%). Deaths were classified as coronary heart disease (CHD), other cardiovascular and non cardiovascular. Among 1063 persons with HF, 45% had preserved EF with less cardiovascular risk factors and less coronary disease than those with reduced EF. At 5 years, survival was 45% (95% CI 43%-49%) and 43% of the deaths were non cardiovascular. The leading cause of death in subjects with preserved EF was non cardiovascular (49%) vs CHD (43%) for subjects with reduced EF. The proportion of cardiovascular deaths decreased from 69% in 1979-1984 to 40% in 1997-2002 (p=0.007) among subjects with preserved EF contrasting with a modest change among those with reduced EF (77% in to 64%, p=0.08). Advanced age, male sex, diabetes, smoking and kidney disease were associated with an increase risk of all cause and cardiovascular death. After adjustment, preserved EF was associated with a lower risk of cardiovascular death but not all cause death. CONCLUSION: Community subjects with HF experience a persistently high mortality and a large proportion of deaths are non-cardiovascular. Subjects with preserved EF have less cardiovascular disease before death, are less likely to experience cardiovascular deaths than those with reduced EF and the proportion of cardiovascular deaths declined over time. PMID- 19300534 TI - Pathological gambling and dopamine agonists: A phenotype? PMID- 19300535 TI - Role of rufinamide in the management of Lennox-Gastaut syndrome (childhood epileptic encephalopathy). AB - Rufinamide, a triazole derivative that is structurally distinct from currently marketed antiepileptic drugs (AEDs), is in development for the adjunctive treatment of Lennox-Gastaut syndrome (LGS) in children and adults. Rufinamide is well absorbed after oral administration, demonstrates low protein binding, and is metabolized by enzymatic hydrolysis without involvement of cytochrome P450 enzymes, conferring a low drug interaction potential. In a randomized, double blind trial involving 138 adult and pediatric patients with LGS, compared with placebo, rufinamide 45 mg/kg/day resulted in significantly superior reductions in drop attacks (median change -42.5% vs +1.4% with placebo) and total seizures ( 32.1% vs -11.7% with placebo), accompanied by significantly higher responder rates. These results are comparable with findings reported for other AEDs in randomized, controlled clinical trials in patients with LGS. Rufinamide produced statistically significant seizure reduction which was maintained during long-term therapy and accompanied by good tolerability. The most frequently reported adverse events from a pooled safety database evaluating short- and long-term therapy were headache (22.9% and 29.5%), dizziness (15.5% and 22.5%) and fatigue (13.6% and 17.7%). Rufinamide therefore presents a favorable efficacy and tolerability profile and is a promising candidate for the adjunctive therapy of LGS. PMID- 19300536 TI - Long-acting risperidone injection: efficacy, safety, and cost-effectiveness of the first long-acting atypical antipsychotic. AB - OBJECTIVE: To review the pharmacokinetics, efficacy, safety, and cost effectiveness of long-acting risperidone. METHODS: Studies published between January 2000 and October 2006 evaluating the pharmacokinetics, efficacy, safety, and cost-effectiveness of long-acting risperidone were reviewed, as identified from literature searches using Medline and EMBASE. Abstracts and posters on long acting risperidone presented at key psychiatry congresses and available in the public domain during this time period were also reviewed. RESULTS: The unique pharmacokinetic profile of long-acting risperidone is derived from the encapsulation of risperidone in a glycolide/lactide matrix in the form of microspheres such that after a single intramuscular injection, significant plasma levels of the drug are achieved after week 3. Steady state, after repeated administration at 2-week intervals, is achieved after 3 injection cycles. Short- and long-term studies have demonstrated that long-acting risperidone (25, 37.5, or 50 mg) is both efficacious and well tolerated in a wide variety of patients with schizophrenia and related psychoses. Most patients can be switched from other oral and long-acting antipsychotic agents without compromising efficacy and safety. Long-acting risperidone may also reduce overall healthcare costs by decreasing rates of relapse and hospitalization. CONCLUSION: The assured delivery of an atypical antipsychotic medication with long-acting risperidone has important implications for patient compliance, maintenance of stability, consistency of treatment, and improving patient outcomes including the achievement of remission. PMID- 19300537 TI - Japanese experience with milnacipran, the first serotonin and norepinephrine reuptake inhibitor in Japan. AB - Milnacipran is a serotonin and norepinephrine reuptake inhibitor (SNRI), with a balanced potency for the inhibition of the reuptake of the two monoamines. In this, it contrasts with venlafaxine and duloxetine which, while possessing a dual action, have a selectivity of the order of 30-fold and 10-fold respectively for the reuptake of serotonin. Milnacipran has mainly been launched in countries where the selective serotonin reuptake inhibitors (SSRIs) and venlafaxine had been established for several years. As such it has attracted relative little interest from clinician investigators as a research tool. Japan, however, represents a unique situation because in 1999 milnacipran was launched within months of the first SSRI and is still the only SNRI in Japan together with only two SSRIs (a third has just been introduced). This has led to a large number of investigative clinical studies, many of which give interesting insights into the potential of milnacipran in the treatment of depression and of other disorders. This article reviews these Japanese studies with milnacipran. PMID- 19300538 TI - Venlafaxine in the treatment of panic disorder. AB - Venlafaxine XR is a novel, dual acting SNRI antidepressant, which inhibits both serotonin and norepinephrine reuptake. Controlled trials have demonstrated the efficacy and safety of venlafaxine in the treatment of panic disorder. Generally well tolerated, with side-effects that usually abate with continued treatment, venlafaxine is an important option to the SSRIs for the treatment of patients with panic disorder. PMID- 19300539 TI - Ziconotide: a review of its pharmacology and use in the treatment of pain. AB - Ziconotide is a powerful analgesic drug that has a unique mechanism of action involving potent and selective block of N-type calcium channels, which control neurotransmission at many synapses. The analgesic efficacy of ziconotide likely results from its ability to interrupt pain signaling at the level of the spinal cord. Ziconotide is a peptidic drug and has been approved for the treatment of severe chronic pain in patients only when administered by the intrathecal route. Importantly, prolonged administration of ziconotide does not lead to the development of addiction or tolerance. The current review discusses the various studies that have addressed the in vitro biochemical and electrophysiological actions of ziconotide as well as the numerous pre-clinical studies that were conducted to elucidate its antinociceptive mechanism of action in animals. In addition, this review considers the pivotal Phase 3 (and other) clinical trials that were conducted in support of ziconotide's approval for the treatment of severe chronic pain and tries to offer some insights regarding the future discovery and development of newer analgesic drugs that would act by a similar mechanism to ziconotide but which might offer improved safety, tolerability and ease of use. PMID- 19300540 TI - Patterns of anterior cingulate activation in schizophrenia: a selective review. AB - BACKGROUND: Anterior cingulate cortex (ACC) dysfunction is implicated in schizophrenia by numerous strands of scientific investigation. Functional neuroimaging studies of the ACC in schizophrenia have shown task-related hypo activation, hyper-activation, and normal activation relative to comparison subjects. Interpreting these results and explaining their inconsistencies has been hindered by our ignorance of the healthy ACC's function. This review aims to clarify the site and magnitude of ACC activations in schizophrenia, and sources of their variation. METHOD: 48 studies of mnemonic and executive task-related activations in schizophrenia using both positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) were analyzed. RESULTS: Abnormal activations in schizophrenia were not restricted to the "cognitive" part of the ACC. Hypoactivations were most common, and were found in all types of tasks. Hyperac-tivations when found, were largely in n-back tasks. CONCLUSIONS: Hypoactivations cannot be explained by poor performance, more demanding control conditions or chronicity of illness alone. Patients on anti-psychotic medication tended to show both greater ACC activation and better performance, although whether this is directly due to their medication or the resultant reduction in symptoms is unclear. The relationship between ACC rCBF and task performance is not straightforward. Future research should better control confounding factors and incorporate different levels of difficulty. PMID- 19300541 TI - Impaired cognition and attention in adults: pharmacological management strategies. AB - Cognitive psychology has provided clinicians with specific tools for analyzing the processes of cognition (memory, language) and executive functions (attention concentration, abstract reasoning, planning). Neuropsychology, coupled with the neurosciences (including neuroimaging techniques), has authenticated the existence of early disorders affecting the "superior or intellectual" functions of the human brain. The prevalence of cognitive and attention disorders is high in adults because all the diseases implicating the central nervous system are associated with cognitive correlates of variable intensity depending on the disease process and the age of the patient. In some pathologies, cognitive impairment can be a leading symptom such as in schizophrenia, posttraumatic stress disorder or an emblematic stigmata as in dementia including Alzheimer's disease. Paradoxically, public health authorities have only recognized as medications for improving cognitive symptoms those with proven efficacy in the symptomatic treatment of patients with Alzheimer's disease; the other cognitive impairments are relegated to the orphanage of syndromes and symptoms dispossessed of medication. The purpose of this review is to promote a true "pharmacology of cognition" based on the recent knowledge in neurosciences. Data from adult human beings, mainly concerning memory, language, and attention processes, will be reported. "Drug therapeutic strategies" for improving cognition (except for memory function) are currently rather scarce, but promising perspectives for a new neurobiological approach to cognitive pharmacology will be highlighted. PMID- 19300542 TI - Managing patient adherence and quality of life in epilepsy. AB - Patient adherence to medication continues to be a cause of concern within the medical profession. This review examines the various methods of quantifying the level of patient adherence, progress in predicting causes of non-adherence, and the implications for its management. Contributions from the medical, health belief, and psychosocial models are discussed in order to highlight how the concept of adherence has changed over time. The impact of epilepsy, seizures, and taking antiepileptic drugs (AEDs) on both adherence and quality of life are also explored. The volume and quality of previous research conducted has enabled a number of predictive factors to be identified, from which various strategies have been developed. While this review concentrates on potential strategies in managing treatment adherence within epilepsy, findings can equally be applied to other chronic conditions. PMID- 19300543 TI - The schizophrenias, the neuroses and the covered wagon; a critical review. AB - OBJECTIVE: In order to compare their validity, this review applies scientific standards for sustaining the neuroses, the schizophrenias and bipolar disorders as separate "bona-fide" psychiatric diseases. The standards for disease validation demand specific and unique symptoms. METHOD: We review a wide variety of clinical and basic science comparisons between schizophrenia and psychotic bipolar in a select English-language literature. RESULTS: Like covered wagons, the neuroses once served us well but became obsolete and were discarded or reorganized based on what was known about commonalities of symptoms, causation and pharmacological responsivity. Bipolar patients meet unique and specific diagnostic criteria and demonstrate consistent results across a variety of scientific disciplines. Neither the neuroses nor the schizophrenias have such unique or disease specific diagnostic criteria. Psychotic mood disorders account for the DSM diagnostic criteria for schizophrenia. A recent, selected but diverse basic science literature demonstrates surprising similarities between schizophrenia and psychotic bipolar which should not exist if these disorders are distinct. CONCLUSIONS: Like the neuroses, there is stigma, confusion and misunderstanding about the condition called schizophrenia, resulting in substantial negative impact on bipolar patients misdiagnosed as having schizophrenia. The psychoses, including the schizophrenias, likely are explained by a single disease, psychotic bipolar disorder, that has demonstrated a wide spectrum of severity of symptoms and chronicity of course, not traditionally recognized. PMID- 19300544 TI - Prodromal non-motor symptoms of Parkinson's disease. AB - The motor symptoms of Parkinson's disease (PD), bradykinesia, muscular rigidity, and tremor depend upon degeneration of the dopaminergic neurons in the substantia nigra pars compacta. Recent neuropathological studies show that the Lewy bodies, the intraneuronal landmark of PD, accumulate in several neuronal cell types in the brain. An ascending gradient of pathological involvement, from the medulla oblongata to neocortical areas has been reported. Thus the original view of PD as a disease characterized by selective damage of the dopaminergic neurons in the mesencephalon should be updated into the concept of a severe multisystemic neurodegenerative disorder. Additionally, the neuropathological alterations outside the substantia nigra are soundly correlated with the non-motor symptoms of PD. As a result of these findings, interest is growing in the identification of prodromal non-motor symptoms of PD. Indeed, data from the literature suggest that autonomic disturbances, olfactory dysfunctions, depression and sleep disorders (in particular REM-sleep behavior disorder) may represent prodromal non motor symptoms of PD. Several tests are available to detect most of these symptoms. Thus, the identification of prodromal non-motor symptoms may contribute to the precocious diagnosis of PD, and might be useful in the future to test the efficacy of neuroprotective agents. PMID- 19300545 TI - One-year follow-up of patients with first-episode schizophrenia (comparison between remitters and non-remitters). AB - Patients admitted to hospital after being diagnosed with first-episode schizophrenia were comprehensively assessed prior to acute treatment (on admission), at the end of the acute treatment (at discharge), and at follow-up after 1 year. The psychopathology was evaluated using the Positive and Negative Syndrome Scale (PANSS). 93 patients were reassessed after 1 year. 73/93 (78%) of the patients fulfilled the criteria for remission. No statistically significant differences in the total PANSS or subscales scores were found between remitters and non-remitters before or after the first episode treatment. However, non remitters had a significantly higher total PANSS score after 1 year than remitters. There was no significant difference in mean psychopathology on admission or at discharge, with the exception of items conceptual disorganization, difficulty in abstract thinking, and lack of judgment and insight between remitters and non-remitters. However, significantly higher mean values were found for all items after 1 year in non-remitters than remitters. On admission the occurrence of positive, negative and general symptoms was balanced; at discharge and after 1 year negative and general symptoms were the most frequently observed. At the 1-year follow-up the impairment of insight and judgment is one of the most frequent symptoms in both remitters (10%) and non remitters (70%). PMID- 19300546 TI - The four As associated with pathological Parkinson disease gamblers: anxiety, anger, age, and agonists. AB - Several studies have related pathological gambling in PD to dopamine agonist therapy. A mail-in survey was sent to PD patients seen at the University of Florida Movement Disorders Center to determine gambling frequency and behavior, and any lifestyle or environmental factors associated with compulsive gambling in PD. 462 surveys were sent and 127 completed surveys were returned, of which ten were from patients who met criteria for compulsive gambling. All ten were taking dopamine agonists coincident with the compulsive gambling. Compulsive gamblers were younger, and psychological distress measures revealed that compulsive gamblers exhibited higher levels of anxiety, anger, and confusion. Thus in this cohort, we have uncovered the several characteristics of the most likely PD compulsive gambler, namely: (young) age, "angry", "anxious", and using a (dopamine) agonist. PMID- 19300547 TI - A survey of five antidepressant properties influencing clinician's treatment choices in MDD. AB - INTRODUCTION: The goal of the present work was to examine how clinicians' perceptions of the properties of antidepressants may influence their choice of antidepressants when treating major depressive disorder (MDD). METHODS: 273 of 682 (40%) clinicians attending a psychopharmacology review course responded to a questionnaire designed to explore their practices and perceptions with regards to antidepressant pharmacotherapy. RESULTS: Most clinicians ranked efficacy (57.3%) as the most important factor when selecting antidepressants, followed by safety (23.0%), tolerability (9.4%), rapidity of action (5.2%), and cost (4.9%). However, when presented with hypothetical scenarios in which there was a difference in efficacy between two antidepressant agents, the relative safety, tolerability, and cost of the two agents significantly influenced the likelihood of choosing one antidepressant over another. In fact, clinicians required progressively greater differences in efficacy between two agents in order to select one antidepressant over another given a difference in terms of their safety than tolerability, or their tolerability than cost (p < 0.0001 all comparisons). CONCLUSIONS: When selecting an antidepressant, clinicians appear to be most influenced by efficacy, followed by safety. Rapidity of action and cost may be less salient considerations in clinical practice. Further research is necessary to elucidate factors that influence clinicians' choice of antidepressants. PMID- 19300548 TI - Aripiprazole treatment of Asperger's syndrome in the acute psychiatric setting: case report. AB - Asperger's syndrome (AS) is under-recognized and may be misdiagnosed as schizophrenia in adults because of symptom overlap. Pharmacological treatment usually targets associated behavioral and mental symptoms rather than the actual core features of AS. We report a middle-aged male patient who, after many years of previous contact with mental health services, and on account of his psychotic symptoms and diagnosis of schizophrenia, was admitted to an inner-city acute psychiatric unit, where a primary diagnosis of AS was established for the first time in his life. His impairing clinical features of AS improved markedly following treatment using aripiprazole, a novel atypical antipsychotic that acts as a partial agonist at dopamine D(2) receptors. As well as sharing clinical features, there is an overlap in underlying neurobiology of AS and schizophrenia, including dopamine dysfunction, that provides a rationale for using antipsychotics of this class in the clinical management not only of associated psychotic symptoms but also of the core features of AS itself. PMID- 19300549 TI - Direct metabolic effects of risperidone and olanzapine in Japanese schizophrenic patients. PMID- 19300550 TI - Why does milnacipran produce so few discontinuation syndromes following abrupt withdrawal? PMID- 19300551 TI - Treatment of generalized anxiety disorder. PMID- 19300552 TI - Role of pregabalin in the treatment of generalized anxiety disorder. AB - Generalized anxiety disorder (GAD) is a common, typically persistent, and disabling condition that is often not recognised, or treated in an evidence-based manner. Current pharmacological and psychological treatment approaches have a number of drawbacks, including a delay in onset of clinical effect, varying relative efficacy against psychological or somatic symptoms of anxiety, potentially troublesome adverse effects, and discontinuation symptoms on stopping treatment. Pregabalin is a structural analog of the inhibitory neurotransmitter gamma amino butyric acid (GABA) but is thought to exert its anxiolytic effects through binding in a state-dependent manner to the alpha-2-delta sub-unit of voltage-gated calcium channels in "over-excited" pre-synaptic neurones, reducing release of excitatory neurotransmitters such as glutamate and substance P. At fixed doses of 200 mg/day or greater, it has consistent proven efficacy in acute treatment of DSM-IV-defined GAD, with some evidence of an early onset of clinical effect, and of efficacy across psychological and somatic anxiety symptom clusters. A pregabalin dosage of 450 mg/day is efficacious in the prevention of relapse. There is at present no published direct comparison with an SSRI. The current known adverse effect profile and studies in healthy volunteers together suggest that pregabalin may have some tolerability advantages over benzodiazepines and venlafaxine, at least in short-term treatment. PMID- 19300553 TI - Duloxetine in the treatment of major depressive disorder. AB - Since depression impacts all body systems, antidepressant treatments should relieve both the emotional and physical symptoms of depression. Duloxetine demonstrated antidepressant efficacy at a dose of 60 mg qd in two placebo controlled, randomized, double-blind studies and significantly improved remission rates compared with placebo. Duloxetine-treated patients had significant reduction in severity of the symptoms of depression as assessed by the HAM-D(17), anxious symptoms as measured by the HAM-A and quality of life measures compared to placebo. Duloxetine also improved somatic symptoms, particularly painful symptoms which may have contributed to significantly improved remission rates compared to placebo. Approximately 10% of the 1139 patients with major depressive disorder in placebo-controlled trials discontinued treatment due to an adverse event, compared to 4% of the 777 patients receiving placebo. In addition to nausea (1.4% incidence), which was the most common reason for discontinuation, dizziness, somnolence, and fatigue were the most common AEs reported as reasons for discontinuation and all were considered drug-related. Duloxetine treatment lacks effects on ECG, increases heart rate, and has little effect on blood pressure or weight. PMID- 19300554 TI - Rivastigmine in the treatment of patients with Alzheimer's disease. AB - Impairment of attention and memory in patients with Alzheimer's disease (AD) is associated with significantly lower levels of acetylcholine. Inhibition of the breakdown of acetylcholine by blocking the enzymes acetylcholinesterase and butyrylcholinesterase with rivastigmine improves this cholinergic depletion. Thus rivastigmine administration provides established, effective, long-term symptomatic treatment in AD and Parkinson's disease (PD) patients with dementia. A sustained treatment with cholinesterase inhibitors in general may also induce a certain deterioration of fine motor behavior, which may play a crucial role in the treatment of PD patients with dementia. Recent studies show that this altered balance between dopamine and acetylcholine due to cholinesterase inhibition, with its possible negative impact on motion behaviour, does not present a major problem in clinical practice in AD patients and may be compensated for by modification of dopaminergic substitution in PD patients with dementia. However, progression of neurodegeneration increases the vulnerability for psychosis in AD and PD patients with dementia in combination with dehydration and often requires additional application of neuroleptics. Since classical neuroleptics increase extrapyramidal symptoms, atypical neuroleptics are used. Out of these, quetiapine shows a distinct lower anticholinergic (muscarinergic) potency with beneficial effects on cognition. This favors its use in combination with rivastigmine. PMID- 19300555 TI - Quetiapine in the treatment of schizophrenia and related disorders. AB - Quetiapine was developed in 1985 by scientists at AstraZeneca (formerly Zeneca) Pharmaceuticals. It received official US Food and Drug Administration approval in September 1997 and approval in Germany in 2000. Since then, quetiapine has been used in the treatment of severe mental illness in approximately 70 countries including Canada, most Western European countries, and Japan. Quetiapine is a dibenzothiazepine derivative with a relatively broad receptor binding profile. It has major affinity to cerebral serotonergic (5HT(2A)), histaminergic (H1), and dopaminergic D(1) and D(2) receptors, moderate affinity to alpha(1)- und alpha(2) adrenergic receptors, and minor affinity to muscarinergic M1 receptors; it demonstrates a substantial selectivity for the limbic system. This receptor occupancy profile with relatively higher affinity for the 5HT(2A) receptor compared with the D(2) receptor is in part responsible for the antipsychotic characteristics and low incidence of extrapyramidal side-effects of quetiapine. The efficacy of quetiapine in reducing positive and negative symptoms of schizophrenia has been proven in several clinical trials with placebo-controlled comparators. Quetiapine has also demonstrated robust efficacy for treatment of cognitive, anxious-depressive, and aggressive symptoms in schizophrenia. Long term trials show sustained tolerability for a broad spectrum of symptoms. Quetiapine has also proven efficacy and tolerability in the treatment of moderate to severe manic episodes, and in the treatment of juveniles with oppositional defiant or conduct disorders, and in the geriatric dementia population. Recent data indicate that quetiapine may also be effective in the treatment of bipolar depressive symptoms without increasing the risk of triggering manic episodes, and in borderline personality disorder. In comparison with other antipsychotics, quetiapine has a favorable side-effect profile. In clinical trials only small insignificant prolongations of the QT interval were observed. Weight-gain liabilities and new-onset metabolic side-effects occupy a middle-ground among newer antipsychotics. As a result of its good efficacy and tolerability profile quetiapine has become well established in the treatment of schizophrenia and manic episodes. PMID- 19300556 TI - The treatment of generalized anxiety disorder with pregabalin, an atypical anxiolytic. AB - A constellation of pharmacologic treatments for generalized anxiety disorder (GAD) have been developed over the past five decades, although each has a number of potential drawbacks in clinical practice. This review addresses one potentially new pharmacologic treatment for generalized anxiety disorder, the gamma-aminobutyric acid analogue pregabalin. We review the mechanism of action, and pharmacokinetic and pharmacodynamic properties of pregabalin as well as the results of 5 double-blind, placebo-controlled trials of pregabalin in the treatment of generalized anxiety disorder (GAD). Based entirely on data from these industry-sponsored (Pfizer), multi-site clinical trials in patients with GAD, pregabalin appears to be generally well tolerated and has rapid onset of action (approximately 1 week), comparable efficacy to benzodiazepines and lower discontinuation rates compared with other pharmacologic treatments. Thus in GAD, a disorder that is often suboptimally responsive to traditional psychotherapeutic and psychopharmacologic interventions - secondary to poor efficacy, tolerability, and/or side-effects - pregabalin may have a primary role in GAD patients, especially in those with certain psychiatric comorbidities or individuals who are on multi-drug regimens for medical comorbidities. PMID- 19300557 TI - Memantine: efficacy and safety in mild-to-severe Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common cause of dementia, accounting for 25 million cases worldwide. Until recently, the pharmacotherapy of AD was limited to the use of cholinesterase inhibitors (ChEIs) that are approved only for the mild to moderate stages of the illness. Memantine, an NMDA receptor antagonist has been found to be effective, both as monotherapy and in combination with donepezil, in the treatment of patients with moderate to severe stage AD. More recent studies have examined the role of memantine in the treatment of the mild to moderate stages of the disease, although the collective results of these studies remain inconclusive. Available pharmacoeconomic data indicate that treatment with memantine is cost-effective when compared with no treatment in patients with moderate to severe AD. Memantine treatment is predicted to be associated with lower costs of care, longer time to dependence and institutionalization, and gains in quality-adjusted life-years. In this article, we review the evidence for the use of memantine in patients with AD, ranging from the mild to severe stages of disease. PMID- 19300558 TI - Glatiramer acetate in the treatment of multiple sclerosis. AB - Glatiramer acetate is an immunomodulating drug used in the treatment of multiple sclerosis. It consists of a copolymer of amino acid residues in the same stoichiometric proportions as in myelin basic protein. Its mechanism of action is not entirely known and is probably multifaceted, with deletion of some immune cell populations and stimulation of others in these patients. Some mechanisms involve neuroprotectant effects. There is ample evidence of its efficacy in relapsing-remitting disease, using both clinical and imaging measures of disease activity, and in this paper we review the clinical and basic studies of this drug. Finally we discuss how some of its neuroprotectant effects may be useful in neurodegeneration such as is seen in more advanced cases of multiple sclerosis and other diseases such as amyotrophic lateral sclerosis and Parkinson's disease. PMID- 19300559 TI - Clinical variables and implications of the personality on the outcome of bipolar illness: a pilot study. AB - Outcome in bipolar patients is affected by comorbidity. Comorbid personality disorders are frequent and may complicate the course of bipolar illness. This pilot study examined a series of 40 euthymic bipolar patients (DSM-IV criteria) (bipolar I disorder 31, bipolar II disorder 9) to assess the effect of clinical variables and the influence of comorbid personality on the clinical course of bipolar illness. Bipolar patients with a diagnosis of comorbid personality disorder (n = 30) were compared with "pure" bipolar patients (n = 10) with regard to demographic, clinical, and course of illness variables. Comorbid personality disorder was diagnosed in 75% of patients according to ICD-10 criteria, with obsessive-compulsive personality disorder being the most frequent type. Sixty three per cent of subjects had more than one comorbid personality disorder. Bipolar patients with and without comorbid personality disorder showed no significant differences regarding features of the bipolar illness, although the group with comorbid personality disorder showed a younger age at onset, more depressive episodes, and longer duration of bipolar illness. In subjects with comorbid personality disorders, the number of hospitalizations correlated significantly with depressive episodes and there was an inverse correlation between age at the first episode and duration of bipolar illness. These findings, however, should be interpreted taking into account the preliminary nature of a pilot study and the contamination of the sample with too many bipolar II patients. PMID- 19300560 TI - An alternative approach to measuring treatment persistence with antipsychotic agents among patients with schizophrenia in the Veterans Health Administration. AB - Prior studies have demonstrated the importance of treatment persistence with anti psychotic agents in sustaining control of schizophrenic symptoms. However, the conventional approach in measuring treatment persistence tended to use only the first prescription episode even though some patients received multiple prescriptions (or multiple treatment episodes) of the same medication within one year following the initiation of the index drug. In this study, we used data from the Veterans Health Administration in the United States to assess the extent to which patients received multiple prescriptions. The study found that about a quarter of the patients had two or more treatment episodes and that levels of treatment persistence tended to vary across treatment episodes. Based on these results, we offered an alternative approach in which we calculated treatment persistence with typical and atypical antipsychotic agents separately for patients with one, two, or three treatment episodes. Considering that patients with different number of treatment episodes might differ in disease profiles, this treatment episode-specific approach offered a fair comparison of the levels of treatment persistence across patients with different number of treatment episodes. Future research needs to extend the analyses beyond two antipsychotic classes to individual antipsychotic agents. A more comprehensive assessment using appropriate analytic methods should help physicians make prescription choices that will ultimately improve the care of patients with schizophrenia. PMID- 19300561 TI - NDT goes Open Access. PMID- 19300562 TI - The role of I-ioflupane SPECT dopamine transporter imaging in the diagnosis and treatment of patients with dementia with Lewy bodies. AB - The diagnosis of dementia with Lewy bodies (DLB) is difficult if one relies solely on clinical features. Current International Consensus Criteria for DLB have high specificity but a significant percentage of patients might be misdiagnosed. Reasons for clinical uncertainty regard the presence of concomitant motor signs in patients with Alzheimer's disease as well as the observation that cognitive abnormalities in DLB might develop with memory impairment without significant parkinsonism. This has clinical relevance as DLB patients may be particularly sensitive to antipsychotics and even the effectiveness of atypical neuroleptics such as quetiapine for the treatment of agitation and hallucinations has been questioned by double-blind, placebo-controlled, randomized studies. By contrast, acetyl-cholinesterase inhibitors such as rivastigmine have shown benefit not only on cognitive but also on psychiatric symptoms. Recent evidence shows that striatal dopamine transporter binding of (123)I-ioflupane SPECT is reduced in DLB and this is consistent with a significant loss of nigral dopamine neurons in this disorder. Several studies have demonstrated the diagnostic accuracy of (123)I-ioflupane in the differential diagnosis of parkinsonism. Given the availability of SPECT, this investigation represents a useful marker to support clinical diagnosis and can help establishing appropriate treatment for this disorder. PMID- 19300563 TI - A review of modafinil film-coated tablets for attention-deficit/hyperactivity disorder in children and adolescents. AB - Modafinil, a wakefulness-promoting agent unrelated to classical sympathomimetic stimulants, has been studied in a total of 933 children and adolescents as a treatment for attention-deficit/hyperactivity disorder (ADHD). Several studies, including three double-blind, placebo-controlled studies with intent-to-treat analyses, have demonstrated the efficacy of modafinil film-coated tablets in reducing symptoms of ADHD and associated problem behaviors in children and adolescents. Modafinil is generally well tolerated, with adverse events (such as insomnia, headache, loss of appetite, weight loss, and gastrointestinal discomfort) that are generally mild to moderate, rarely leading to medication discontinuation. To minimize treatment-emergent side effects, titration to the target dose of 355-425 mg once a day should take place over 2-3 weeks. Due to reports of skin rash (including one case of possible erythema multiforme/Stevens Johnson Syndrome during pivotal studies), additional studies have been requested to better evaluate the risks of developing severe cutaneous adverse reactions. PMID- 19300565 TI - Continuous levodopa for advanced Parkinson's disease. AB - Parkinson's disease is characterized by the progression of the disease from the early stages where it still has little functional consequence for afflicted patients, to an advanced stage disease with large consequences in terms of function, quality of life and individual and societal costs. Motor fluctuations and symptoms of levodopa overdosage may occur in parallel with increasing Parkinsonian symptoms. This leads to a narrower therapeutic window which causes problems with traditional oral medication. Various ways of optimizing oral treatment should be tried but often have limited effects. In addition to the previous alternatives of neurosurgery (especially deep brain stimulation of the subthalamic nuclei) and continuous apomorphine treatment there is now also the alternative of continuous enteral levodopa administration via a trans-abdominal tube. The effect of the treatment may be tested individually via naso-duodenal administration before a decision is made whether to continue with permanent treatment. In the present article, the challenges to treatment of Parkinson's disease in these phases are described as well as the various treatment alternatives available. Focus is mainly on the clinical studies of continuous levodopa infusion therapies, especially enteral administration of levodopa/carbidopa gel. The place of enteral levodopa/carbidopa gel treatment among the other treatment methods is also discussed. PMID- 19300564 TI - Donepezil in Alzheimer's disease: From conventional trials to pharmacogenetics. AB - Donepezil is the leading compound for the treatment of Alzheimer's disease (AD) in more than 50 countries. As compared with other conventional acetylcholinesterase inhibitors (AChEIs), donepezil is a highly selective and reversible piperidine derivative with AChEI activity that exhibits the best pharmacological profile in terms of cognitive improvement, responders rate (40% 58%), dropout cases (5%-13%), and side-effects (6%-13%) in AD. Although donepezil represents a non cost-effective treatment, most studies convey that this drug can provide a modest benefit on cognition, behavior, and activities of the daily living in both moderate and severe AD, contributing to slow down disease progression and, to a lesser exetnt, to delay institutionalization. Patients with vascular dementia might also benefit from donepezil in a similar fashion to AD patients. Some potential effects of donepezil on the AD brain, leading to reduced cortico-hippocampal atrophy, include the following: AChE inhibition, enhancement of cholinergic neurotransmission and putative modulation of other neurotransmitter systems, protection against glutamate-induced excitotoxicity, activation of neurotrophic mechanisms, promotion of non-amyloidodgenic pathways for APP processing, and indirect effects on cerebrovascular function improving brain perfusion. Recent studies demonstrate that the therapeutic response in AD is genotype-specific. Donepezil is metabolized via CYP-related enzymes, especially CYP2D6, CYP3A4, and CYP1A2. Approximately, 15%-20% of the AD population may exhibit an abnormal metabolism of AChEIs; about 50% of this population cluster would show an ultrarapid metabolism, requiring higher doses of AChEIs to reach a therapeutic threshold, whereas the other 50% of the cluster would exhibit a poor metabolism, displaying potential adverse events at low doses. In AD patients treated with a multifactorial therapy, including donepezil, the best responders are the CYP2D6-related extensive (EM)(*1/*1, *1/*10) (57.47%) and intermediate metabolizers (IM)(*1/*3, *1/*5, *1/*6, *7/*10) (25.29%), and the worst responders are the poor (PM) (*4/*4)(9.20%) and ultra-rapid metabolizers (UM) (*1xN/*1) (8.04%). Pharmacogenetic and pharmacogenomic factors may account for 75%-85% of the therapeutic response in AD patients treated with donepezil and other AChEIs metabolized via enzymes of the CYP family. The implementation of pharmacogenetic protocols can optimize AD therapeutics. PMID- 19300566 TI - Mechanisms of modafinil: A review of current research. AB - The novel wake-promoting agent modafinil has been in use for the treatment of several sleep disorders for a few years and is now undergoing clinical trials for its use in the treatment of stimulant addiction, but its primary mechanism of action remains elusive. Previous laboratory studies have shown that modafinil has antioxidative and neuroprotective effects, which have not previously been suggested to be related to its wake-promoting effects. However, recent research indicates that free radicals may be related to sleep induction as well as cellular damage, suggesting that a common target of action may mediate modafinil's ability to oppose both of these effects. In this review we summarize and discuss previously published research on modafinil's neural, cytoprotective, and cognitive effects, and we propose possible primary biochemical targets that could underlie the effects of modafinil observed in these studies. We also suggest neurocognitive mechanisms responsible for modafinil's cognitive enhancing effects and its therapeutic potential in the treatment of stimulant addiction. PMID- 19300567 TI - Mechanisms of over-activated innate immune system regulation in autoimmune and neurodegenerative disorders. AB - Reactions of innate immunity include phagocytosis, the production and activity of cytokines, chemokines, and adhesion molecules, the killing of infected or changed cells by NK cells and complement activated by natural lectins, and the cytokine dependent resistance of leukocytes to viral infection. All these mechanisms maintain innate immunity. Deficiency in this immunity is sometimes accompanied by frequent bacterial and viral infections. When innate immunity is permanently stimulated and the intensity of the reactions is stronger, these mechanisms may be directed against the host and subsequently stimulate acquired immunity (antibody and cellular immunity). A higher production of cytokines, oxidative stress, and a high production of NO accompany autoimmunity and neurodegeneration. The possible participation of innate immune receptors, cytokines, and other factors in the development of autoimmune and neurodegenerative diseases is discussed. The importance and possible role of blood-derived microglial cells in the prevention or elimination of amyloid deposits and plaque formation is described. A possible regulatory system, based on the presence of suppressors of cytokine signaling (SOCS), receptors of the Tyro-3 family, adenosine and adenosine phosphates, and IL-10, is reviewed. This review presents the mechanisms involved in the control of the innate immune response by microglia in the development of neurodegenerative disorders. PMID- 19300568 TI - Development and psychometric properties of the Patient-Head Injury Participation Scale (P-HIPS) and the Patient-Head Injury Neurobehavioral Assessment Scale (P HINAS): patient and family determined outcomes scales. AB - OBJECTIVE: To develop a measure to assess post-acute outcome following from traumatic brain injury (TBI) with particular emphasis on the emotional and the behavioral outcome. The second objective was to assess the test-retest reliability, internal consistency, and factor structure of the newly developed patient version of the Head Injury Participation Scale (P-HIPS) and Patient-Head Injury Neurobehavioral Scale (P-HINAS). METHOD: Thirty-two TBI individuals and 27 carers took part in in-depth qualitative interviews exploring the consequences of the TBI. Interview transcripts were analyzed and key themes and concepts were used to construct the 49-item P-HIPS. A postal survey was then conducted on a cohort of 113 TBI patients to 'field test' the P-HIPS and the P-HINAS. RESULTS: All individual 49 items of the P-HIPS and their total score showed good test retest reliability (0.93) and internal consistency (0.95). The P-HIPS showed a very good correlations with the Mayo Portland Adaptability Inventory-3 (MPAI-3) (0.87) and a moderate negative correlation with the Glasgow Outcome Scale Extended (GOSE) (-0.51). Factor analysis extracted the following domains: 'Emotion/Behavior,' 'Independence/Community Living,' 'Cognition' and 'Physical'. The 'Emotion/Behavior' factor constituted the P-HINAS, which showed good internal consistency (0.93), test-retest reliability (0.91) and concurrent validity with MPAI subscale (0.82). CONCLUSIONS: Both the P-HIPS and the P-HINAS show strong psychometric properties. The qualitative methodology employed in the construction stage of the questionnaires provided good evidence of face and content validity. PMID- 19300569 TI - Development and psychometric properties of the Carer - Head Injury Neurobehavioral Assessment Scale (C-HINAS) and the Carer - Head Injury Participation Scale (C-HIPS): patient and family determined outcome scales. AB - OBJECTIVE: Develop and assess the psychometric properties of the Carer - Head Injury Participation Scale (C-HIPS) and its biggest factor the Carer - Head Injury Neurobehavioral Assessment Scale (C-HINAS). Furthermore, the aim was to examine the inter-informant reliability by comparing the self reports of individuals with traumatic brain injury (TBI) with the carer reports on the C HIPS and the C-HINAS. METHOD: Thirty-two TBI individuals and 27 carers took part in in-depth qualitative interviews exploring the consequences of the TBI. Interview transcripts were analysed and key themes and concepts were used to construct a 49-item and 58-item patient (Patient - Head Injury Participation Scale [P-HIPS]) and carer outcome measure (C-HIPS) respectively, of which 49 were parallel items and nine additional items were used to assess carer burden. Postal versions of the P-HIPS, C-HIPS, Mayo Portland Adaptability Inventory-3 (MPAI-3), and the Glasgow Outcome Scale-Extended (GOSE) were completed by a cohort of 113 TBI individuals and 80 carers. Data from a sub-group of 66 patient/carer pairs were used to compare inter-informant reliability between the P-HIPS and the C HIPS, and the P-HINAS and the C-HINAS respectively. RESULTS: All individual 49 items of the C-HIPS and their total score showed good test-retest reliability (0.95) and internal consistency (0.95). Comparisons with the MPAI-3 and GOSE found a good correlation with the MPAI-3 (0.7) and a moderate negative correlation with the GOSE (-0.6). Factor analysis of these items extracted a 4 factor structure which represented the domains 'Emotion/Behavior' (C-HINAS), 'Independence/Community Living', 'Cognition', and 'Physical'. The C-HINAS showed good internal consistency (0.92), test-retest reliability (0.93), and concurrent validity with one MPAI subscale (0.7). Assessment of inter-informant reliability revealed good correspondence between the reports of the patients and the carers for both the C-HIPS (0.83) and the C-HINAS (0.82). CONCLUSION: Both the C-HINAS and the C-HIPS show strong psychometric properties. The qualitative methodology employed in the construction stage of the questionnaires provided good evidence of face and content validity. Comparisons between the P-HIPS and the C-HIPS, and the P-HINAS and the C-HINAS indicated high levels of agreement suggesting that in situations where the patient is unable to provide self-reports, information provided by the carer could be used. PMID- 19300570 TI - Levetiracetam in the treatment of childhood epilepsy. AB - Epilepsy is a common pediatric neurologic disorder that is difficult to manage in a substantial portion of children. Levetiracetam (LEV) is a novel antiepileptic drug (AED) that has recently been approved as add-on treatment for various seizure types in epilepsy populations that include children: for refractory partial seizures in epilepsy patients >/=4 years old, for myoclonic seizures in juvenile myoclonic epilepsy patients >/=12 years old, and for primary generalized tonic-clonic seizures in idiopathic generalized epilepsy patients (>/=6 years old with FDA approval; >/=12 years old with EMEA approval). A review of published pediatric studies indicates that the efficacy of LEV is best established for partial seizures; however, results from recent double-blind and open-label trials indicate that adjunctive LEV also controls generalized seizures - particularly myoclonic and generalized tonic-clonic - in children and adolescents with primary generalized epilepsy. LEV was well-tolerated in pediatric studies. The most common adverse events (AEs) reported were sedation related. Behavioral AEs were among the most commonly reported events in some trials; conversely, improvements in behavior and cognition were also frequently reported. LEV appears to be a safe and effective AED with unique characteristics that benefit the treatment of children with epilepsy. PMID- 19300571 TI - Agomelatine and its therapeutic potential in the depressed patient. AB - Despite advances in understanding potential disease mechanisms and in developing novel therapeutic approaches to the treatment of major depressive disorder, the disease continues to carry an enormous personal, social, and economic burden. Agomelatine represents an important opportunity to advance the treatment of depression. It is a melatonergic (MT(1) and MT(2)) agonist and serotonergic (5HT(2C)) antagonist. Evidence from animal models of depression, complements emerging clinical data. In a dose range of 25-50 mg daily, agomelatine is an effective antidepressant with a very favorable side-effect profile. In particular, sleep restorative action in the absence of sedation and minimal effect on sexual function suggests that agomelatine represents a worthwhile treatment alternative for patients with major depressive disorder. PMID- 19300572 TI - Betahistine in the treatment of Meniere's disease. AB - Meniere's disease and related disease of the vestibular system are common and debilitating. Current therapy is multi-modal and includes drug therapy and lifestyle adaptations. Unfortunately many of the drugs used in treatment (particularly those used to control nausea) are sedative and hamper the process of vestibular compensation. Although betahistine (Serc(r)), BetaSerc(r)); Solvay Pharmaceuticals) is the mainstay of drug treatment in these illnesses, its efficacy has not, until recently, been evaluated to modern standards. Betahistine is an analog of histamine with weak agonist properties at histamine H1 receptors and more potent anatgonistic effects at histamine H3 receptors. Growing evidence suggests that the mechanism of action of betahistine lies in the central nervous system and in particularly in the neuronal systems involved in the recovery from process after vestibular loss. The histaminergic neurones of the tuberomamillary and vestibular nuclei are implicated. In recent years the clinical efficacy of betahistine has been demonstrated in double-blind, randomized, placebo, and active controlled studies in adequate numbers of patients. Although the results of comparative studies between betahistine and other drugs (flunarizine, cinnarizine, and cinnarizine + dimenhydrate) are equivocal, the efficacy of betahistine is now clear. PMID- 19300573 TI - Eszopiclone: its use in the treatment of insomnia. AB - Eszopiclone is the S-isomer of racemic zopiclone, a cyclopyrrolone with sedative hypnotic activity that has been available in Europe, Canada, and Latin America since 1987. Eszopiclone acts by binding to the GABA(A) receptor. In contrast to the benzodiazepine (BZD) hypnotics, eszopiclone has more selectivity for certain subunits of the GABA(A) receptor. Oral eszopiclone is rapidly absorbed and extensively distributed to body tissues including the brain. Peak plasma concentrations are attained 1.0-1.6 hours after a 3 mg dose, while the mean elimination half-life is 6 hours. The half-life increases with age to about 9.0 hours in patients 65 years or older. Eszopiclone's pharmacokinetic (PK) profile is not substantially modified in patients suffering from renal failure or mild-to moderate hepatic impairment, although patients with severe hepatic insufficiency should have a reduced dose. The subjective perception of improved sleep following eszopiclone 2 or 3 mg treatment has been demonstrated in randomized, double blind, placebo-controlled studies of up to 6 months' duration. In these studies the drug significantly reduced sleep onset latency (SOL), the number of awakenings, and wake time after sleep onset (WASO) whereas total sleep time (TST) and quality of sleep were increased in non-elderly and elderly subjects. Sleep laboratory studies of the effects of eszopiclone have confirmed the drug's clinical efficacy in subjects with chronic primary insomnia. Eszopiclone, unlike BZD hypnotics, does not significantly alter values corresponding to slow wave sleep (SWS or stages 3 and 4) and rapid eye movement (REM) sleep. Rebound insomnia following withdrawal of eszopiclone has been examined in only one study. Discontinuation of the active treatment with 2 mg was followed by rebound insomnia in non-elderly subjects. Three-mg doses of eszopiclone administered for a period of up to 12 months was associated with a sustained beneficial effect on sleep induction and maintenance, with no occurrence of tolerance. The most common side-effects were unpleasant or bitter taste, headache, dyspepsia, pain, diarrhea, dry mouth, upper respiratory infection, urinary tract infection, dizziness, and accidental injury. New adverse events (withdrawal symptoms) including anxiety, abnormal dreams, hyperesthesia, nausea, and upset stomach were recorded in one study on the days following eszopiclone 2 or 3 mg discontinuation. Although dependence and abuse potential have not been formally assessed, unpublished data show that eszopiclone at doses of 6 and 12 mg produces euphoria effects similar to those of diazepam 20 mg in BZD drug addicts. In conclusion, available evidence tends to indicate that eszopiclone is effective and safe for the treatment of chronic primary insomnia in non-elderly and elderly subjects. Tolerance did not occur during active drug administration for a 12 month period. Thus eszopiclone can be efficacious not only during short- and intermediate-term administration but also in patients requiring prolonged regular drug usage. PMID- 19300574 TI - An emerging role for escitalopram in the treatment of obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is a common and severe neuropsychiatric disorder treated by both behavioral and pharmacologic techniques. Despite the availability of treatments for OCD, including the selective serotonin reuptake inhibitors (SSRIs), many OCD patients have an inadequate response to current treatments. As such, additional approaches to the management of OCD are required. A potential but little studied treatment for OCD is the SSRI escitalopram. Escitalopram is the S-enantiomer of citalopram, the preparation containing both S and R enantiomers of citalopram. Not only is escitalopram the most selective of the SSRIs, it is also devoid of R-citalopram, which may interfere with the effects of the S enantiomer. Escitalopram appears to be effective in depression and several anxiety disorders, including social anxiety disorder and generalized anxiety disorder, conditions in which it also appears reasonably well tolerated. Enantiomeric specificity, high serotonin reuptake selectivity, comparatively good tolerability and favorable pharmacokinetics, and preliminary evidence of efficacy in OCD suggest a potential role for the use of escitalopram in the treatment of OCD. Nevertheless, additional work including evaluating the use of escitalopram with behavioral interventions and in long-term treatment of OCD is needed to clarify its overall role in managing OCD. PMID- 19300575 TI - The role of lamotrigine in the management of bipolar disorder. AB - Lamotrigine has emerged with a distinct place in the pharmacological treatment of bipolar disorder, with the potential to treat and prevent bipolar depression, which is the dominant and arguably most disabling and under-treated phase of the illness. This review examines the published clinical trials of lamotrigine in bipolar treatment. While the data supports its tolerability and safety, the strongest evidence for its efficacy lies in the prevention of bipolar depression, with weaker evidence for the treatment of acute bipolar depression, refractory unipolar and bipolar depression, and rapid cycling bipolar disorder. The total number of published well designed trials is small, even the maintenance evidence is derived from two studies. However, this relative inadequacy compares favorably with the alternative treatment options for bipolar depression, which are marked by poor efficacy or risk of polarity switch. The designation of lamotrigine as first-line treatment for bipolar depression prophylaxis should be done in cognizance of this context, and it would seem prudent to await greater evidence of efficacy before designating lamotrigine as first-line treatment for other bipolar indications. Further randomized controlled trials are required to consolidate the available findings and to explore the boundaries of lamotrigine's efficacy, which may encompass the soft spectral disorders. PMID- 19300576 TI - Novel research translates to clinical cases of schizophrenic and cocaine psychosis. AB - Pharmacotherapies for schizophrenic and cocaine psychoses are complex but similar because of similarities in their brain neurochemistry and behavioral outcomes. Their neurochemical neuronal mechanisms of action, as shown in preclinical and clinical studies, involve primarily dopaminergic dysfunction and, secondarily, neuroadaptive effects that seem to involve central serotonergic function. Behavioral outcomes of both disorders include hyperactivity and antipsychotic medications can ameliorate psychotic symptoms. Patients with both disorders often arrive at emergency departments and present floridly psychotic with a predominance of positive symptoms, often prompting physicians to select a typical antipsychotic medication such as haloperidol. While this has become conventional wisdom, we believe that to use an atypical antipsychotic medication, such as risperidone, in the treatment of both psychoses is quite rational for long-term management of both positive and negative symptoms. Also, controlled clinical studies have shown that risperidone, an atypical antipsychotic medication, is successful in the treatment of cocaine dependence and withdrawal (Smelson et al 1997, 2002; Grabowski et al 2000). Furthermore, the availability and effectiveness of long-acting risperidone in injectable form opens new possibilities for the long-term management of both disorders. In this paper, we present data which show that the use of risperidone is plausible for effective pharmacotherapy of schizophrenic and cocaine psychoses. PMID- 19300577 TI - Depression and smoking cessation: does the evidence support psychiatric practice? AB - Depression and smoking are highly comorbid. The vast majority of psychiatrists treating depressed patients do not target or treat nicotine dependence, and many inpatient psychiatric facilities implicitly condone smoking by providing 'smoke breaks'. The reasons for failure to treat are unclear, but are probably linked to the notion that depressed smokers are neither willing nor able to quit, and will become more depressed if they try. We review the clinical evidence on depression and smoking cessation, and find little support for current psychiatric practice. Although quitting smoking does appear to pose a risk for the development of depression, this risk is not clearly higher in those with a past history of depression than those without. Depressed smokers are as capable as nondepressed smokers of quitting smoking, and at least one-quarter of depressed smokers is willing to try. Sustained abstinence may even lead to improvement in depressive disorders. More research is needed to understand the relationship between depression and quitting smoking, but current clinical evidence suggests more resiliency among depressed smokers than common clinical wisdom would dictate. PMID- 19300578 TI - Fifty years chlorpromazine: a historical perspective. AB - Chlorpromazine was synthesized in December 1951 in the laboratories of Rhone Poiulenc, and became available on prescription in France in November 1952. Its effectiveness was reflected in the transformation of disturbed wards; its commercial success stimulated the development of other psychotropic drugs. Recognition of chemical mediation at the site of the synapse, followed by the introduction of the spectrophotofluorimeter first, and receptor assays subsequently, led to the demonstration that chlorpromazine blocks dopamine receptors. Treatment with chlorpromazine focused attention on the heterogeneity of schizophrenia in terms of responsiveness to treatment. By the mid-1980s there was sufficient evidence to believe that resolving this heterogeneity is a prerequisite for developing more effective treatments. Chlorpromazine was instrumental in the development of neuropsychopharmacology, a new discipline dedicated to the study of mental pathology with the employment of centrally acting drugs. PMID- 19300579 TI - New assessment tools that measure sleep vital signs: the SleepMed Insomnia Index and the Sleep Matrix. AB - Insomnia is the leading sleep disorder in the US; however, diagnosis is often problematic. This pilot study assessed the clinical value of a novel diagnostic insomnia questionnaire. The SleepMed Insomnia Index (SMI) was administered to 543 consecutive patients and 50 normal control subjects during a pilot study. Mean SMI scores were assessed based on subsequent sleep-related diagnoses. The SMI scores for patients with sleep-related disorders were significantly higher than those for the control group (p < 0.001) and highest for the 90 patients comprising the insomnia group. Analysis of the SMI scores from the 90 insomnia patients indicates a high degree of reliability (Cronbach's alpha: 0.7). These data support our clinical experience with this diagnostic tool which indicates a strong likelihood of disrupted nighttime sleep in patients with high SMI scores. Following further validation, the SMI may prove to be a valuable tool for evaluating sleep disorders, specifically as an aid in the diagnosis of insomnia. The Sleep Matrix is a visual tool that quantifies a sleep complaint by combining scores from the Epworth Sleepiness Scale (ESS) and the SMI. The SMI measures an insomnia component while the ESS is an accepted measure of daytime sleepiness. The Sleep Matrix visually displays the complexity of the sleep complaint in an effort to differentiate insomnia with differing etiologies from other sleep disorders and measure treatment outcomes. To pilot test the Sleep Matrix, the tool was administered to 90 patients with insomnia and to 22 normal controls. Plots from the insomnia patients were concentrated into the "insomnia zone" while scores from the normal controls were located in the "normal zone" located in the lower left quadrant. Additional research using the Sleep Matrix could provide data that the tool could be utilized to visually aid the clinician in the diagnosis of unknown sleep complaints. PMID- 19300580 TI - Patients' health literacy in psychotic disorders. AB - Compliance and relapse are major issues in the treatment of psychotic disorders. About 50% of subjects with schizophrenia do not comply with treatment and relapse rates of 65% are reported after one year and 80% after two years. Drug treatments are effective against psychotic symptoms, but cannot promote functional recovery or prevent relapses when prescribed alone. The factors influencing compliance include side effects and the patients' awareness of their illness. Psychosocial interventions, cognitive remediation and psychotherapy have been proposed as adjuvant treatments to increase compliance and to decrease the rate of relapse. Most of these interventions have been shown to increase compliance and to decrease the rate of relapse, but the most robust results have been achieved with cognitive behavioral therapy. PMID- 19300581 TI - New antidepressants or more of the same? PMID- 19300582 TI - Zolpidem modified-release in insomnia. AB - Zolpidem modified-release (MR) is the first hypnotic agent to be marketed in an extended-release formulation. Zolpidem MR is a two-layered, biphasic release tablet indicated for the management of induction of sleep and sleep maintenance. The pharmacokinetics of the drug are similar to those of immediate-release zolpidem. Two double-blind, placebo-controlled, parallel-group trials demonstrated efficacy in adults and elderly patients treated with zolpidem MR for 3 weeks without significant impairment in next-day psychomotor functioning. The most common adverse effects with zolpidem MR were dizziness, somnolence, and headache. A starting dose of zolpidem MR 12.5 mg is recommended for adults and 6.25 mg for elderly patients. PMID- 19300583 TI - Transdermal selegiline for the treatment of major depressive disorder. AB - Non-selective inhibition of monoamine oxidase (MAO) enzymes (ie, isoforms A and B) in the brain are associated with clinically significant antidepressant effects. In the US, the selegiline transdermal system (STS; EMSAM) is the first antidepressant transdermal delivery system to receive Food and Drug Administration (FDA) approved labeling for the treatment of major depressive disorder (MDD). Currently, the use of orally administered MAO inhibitor antidepressants (eg, phenelzine, tranylcypromine) is limited by the risk of tyramine-provoked events (eg, acute hypertension and headache, also known as the "cheese reaction") when combined with dietary tyramine. The selegiline transdermal system is the only MAOI available in the US for the treatment of MDD that does not require dietary restriction at the clinically effective dose of 6 mg/24 hours. Delivery of selegiline transdermally (EMSAM((R))) bypasses hepatic first pass metabolism, thereby avoiding significant inhibition of gastrointestinal and hepatic MAO-A activity (ie, reduced risk of tyramine provoked events) while still providing sufficient levels of selegiline in the brain to produce an antidepressant effect. At dosages of 6-12 mg/24 hours, EMSAM has been shown to improve symptoms of depression, have good tolerability, and have high rates of medication adherence. However, at higher doses of EMSAM (ie, 9 mg/24 hours or more), dietary restriction of tyramine intake is recommended. The introduction of EMSAM overcomes many of the safety concerns affiliated with the conventional oral MAO inhibitors and EMSAM may be considered another strategy for the treatment of MDD, especially in patients who cannot tolerate oral antidepressants, are poorly adherent, who present with atypical depressive symptoms, or have failed other antidepressants. PMID- 19300584 TI - Update on the management of neuromuscular block: focus on sugammadex. AB - Steroidal neuromuscular blocking agents (NMBAs), such as rocuronium, are widely used in clinical anesthesia and emergency medicine to facilitate endotracheal intubation and artificial ventilation and to allow surgical access to body cavities. Reversal of neuromuscular blockade is important for the acceleration of patient recovery and prevention of postoperative residual neuromuscular blockade and reduces the incidence of severe morbidity and mortality associated with anesthesia management. Sugammadex is the first selective relaxant binding agent (SRBA) and has been designed to reverse the steroidal neuromuscular blocking drug rocuronium. Encapsulation of the rocuronium molecule by sugammadex results in a rapid decrease in free rocuronium in the plasma and subsequently at the nicotinic receptor at the motor endplate. After encapsulation, rocuronium is not available to bind to the nicotinic receptor in the neuromuscular junction. This promotes the liberation of acetylcholine receptors, and muscle activity reappears. This new concept of reversal of neuromuscular block induced by rocuronium (or vecuronium) led to impressive results in animal and phase 1 and 2 studies. Sugammadex is currently in phase 3 clinical studies and may be commercially available by 2008. PMID- 19300585 TI - Sleep deprivation: Impact on cognitive performance. AB - Today, prolonged wakefulness is a widespread phenomenon. Nevertheless, in the field of sleep and wakefulness, several unanswered questions remain. Prolonged wakefulness can be due to acute total sleep deprivation (SD) or to chronic partial sleep restriction. Although the latter is more common in everyday life, the effects of total SD have been examined more thoroughly. Both total and partial SD induce adverse changes in cognitive performance. First and foremost, total SD impairs attention and working memory, but it also affects other functions, such as long-term memory and decision-making. Partial SD is found to influence attention, especially vigilance. Studies on its effects on more demanding cognitive functions are lacking. Coping with SD depends on several factors, especially aging and gender. Also interindividual differences in responses are substantial. In addition to coping with SD, recovering from it also deserves attention. Cognitive recovery processes, although insufficiently studied, seem to be more demanding in partial sleep restriction than in total SD. PMID- 19300586 TI - Progress update: Pharmacological treatment of Alzheimer's disease. AB - A number of drugs have been approved for the treatment of Alzheimer's disease (AD) and a larger number are being studied as possible therapies. The current mainstays of the pharmacotherapy of AD are the cholinesterase inhibitors (donepezil, galantamine, rivastigmine) and memantine. They collectively have acceptable tolerability and proven but modest efficacy. The agents being studied include dietary supplements (eg, vitamin E), herbal preparations (eg, Ginkgo biloba), medications approved for other indications (eg, HMG-CoA reductase enzyme inhibitors) and research drugs. In this review we discuss in detail the approved agents and review a number of the unapproved therapies that are currently available to the practitioner. While our era offers much more in the way of therapeutics for AD, it is clear that more work still needs to be done. PMID- 19300587 TI - Review of olanzapine in the management of bipolar disorders. AB - Olanzapine is an atypical antipsychotic currently with indications for the treatment of schizophrenia, acute mania and the prevention of relapse in bipolar disorder. A growing body of clinical evidence supports these indications. Acute mania trials have demonstrated superior efficacy of olanzapine to placebo, equal or superior efficacy to valproate and superior efficacy in combination therapy with lithium or valproate compared to mood stabilizer monotherapy. Olanzapine demonstrated a modest effect in the treatment of bipolar depression with a substantially enhanced effect in combination with fluoxetine. Maintenance trials showed olanzapine to be more efficacious than placebo in the prevention of manic and depressive relapses and non-inferior to lithium or valproate. Combination of olanzapine with lithium or valproate was also found to be more efficacious than lithium or valproate monotherapy in the prevention of manic relapse in patients with a partial response to monotherapy with lithium or valproate. These trials suggest that olanzapine is a viable option and an invaluable addition to the pharmacological armamentarium in the treatment of bipolar I disorder. However, this can often be mitigated by safety and tolerability concerns with this agent including weight gain and metabolic syndrome that warrants clinician vigilance and discernment that is imperative in today's clinical practice. PMID- 19300588 TI - Late-onset Diogenes syndrome in Chinese - an elderly case series in Hong Kong. AB - We review a consecutive case series of elders presenting to a regional psychogeriatric service in Hong Kong in 1996-2001. Eighteen elders (aged 65 and over) fulfilled the classical symptoms of Diogenes syndrome (extreme squalor, neglected physical state, unhygienic condition & social isolation with or without hoarding). A diverse clinical and socio-demographic profile was observed. Most of our clients suffered from different stages of dementia. Other diagnoses such as schizophrenia and alcohol abuse were diagnosed in this cohort as comorbid or independent conditions. Neither psychopathology nor social situations could adequately account for the initiation and perpetuation of Diogenes syndrome in some cases. The plausible psychological etiologies are discussed in the context of existential values in Chinese culture and changes in traditional family dynamics as the society modernizes. PMID- 19300589 TI - The amyloid beta ion channel hypothesis of Alzheimer's disease. AB - Alzheimer's disease (AD) is a leading cause of chronic dementia in the US. Its incidence is increasing with an attendant increase in associated health care costs. Since its first description in a patient by Dr. Alois Alzheimer over a century ago, a large body of biomedical literature has established a detailed clinical and molecular profile of this disorder. Amyloid beta peptide (Abeta; a 39-42 amino acid molecule) is the major component of senile plaques, the lesions that are one of the pathologic hallmarks of AD (Wong et al 1985). Although many aspects of the biology of amyloid beta have been investigated, several fundamental questions about how this peptide causes AD neuropathology remain unanswered. The key question is: How is Abeta toxic to cerebral neurons? Because plaques are extra-neuronal deposits, it is difficult to imagine a structural basis for their toxicity. As an interesting contrast the other pathognomonic feature of AD, neurofibrillary tangles, are intra-axonal structural anomalies that are composed of the hyperphosphorylated microtubule associated (MAP) protein, tau. This review will assess the current thinking that relates to a recent hypothesis of Abeta toxicity. In 1992, Hardy and Higgins reported findings that suggested a new and intriguing possibility. These authors found that Abeta peptides disrupt Ca(2+) homeostasis in neurons and increase intracellular Ca(2+) [Ca(2+)](i). This was corroborated by Mattson and his colleagues who demonstrated that Abeta exposure to human cortical neurons raised [Ca2(+)](i) (Mattson, Cheng et al 1992); (Hardy and Higgins 1992). Finally, Nelson Arispe's group at the NIH specifically investigated the possibility that Abeta peptides might function like Ca(2+) ion channels (Arispe et al 1993). This and several subsequent studies have laid the foundation for a novel idea: "Abeta peptides are, in part, toxic to neurons because they form aberrant ion channels in neuronal membranes and thereby disrupt neuronal homeostasis". In this review we shall critically examine this theory in light of classic and contemporary literature. PMID- 19300590 TI - Whole genome association studies of neuropsychiatric disease: An emerging era of collaborative genetic discovery. AB - Family history, which includes both common environmental and genetic effects, is associated with an increased risk for many neuropsychiatric diseases. Investigators have identified several disease-causing mutations for specific neuropsychiatric disorders that display Mendelian segregation. Such discoveries can lead to more rational drug design and improved intervention from a better understanding of the underlying biological mechanisms. However, a key challenge of genetic discovery in human complex diseases, including neuropsychiatric disorders, is that most diseases with genetic components display non-Mendelian patterns of inheritance. Recent advances in human population genetics include high-density genome-wide analyses of single nucleotide polymorphisms (SNPs) that make it possible to study complex genetic contributions to human disease. This approach is currently the most powerful strategy for analyzing the genetics of complex diseases. Genome-wide SNP analyses often require a large collaborative effort to collect, manage, and disseminate the numerous samples and corresponding clinical data. In this review we discuss the use of publicly available biorepositories for the collection and distribution of human genetic material, associated phenotypic information, and their use in genome-wide investigations of human neuropsychiatric diseases. PMID- 19300591 TI - Dementia with Lewy bodies: Definition, diagnosis, and pathogenic relationship to Alzheimer's disease. AB - Clinical dementia associated with the appearance of Lewy bodies in the cerebral cortex has been recognized for over 40 years. Until the 1990s, however, cortical Lewy body disease was thought to be a rare cause of dementia. At that time, the advent of sensitive and specific immunohistochemical techniques for highlighting these elusive structures led to the recognition of cortical Lewy body disease as a common substrate for clinical dementia. Current diagnostic criteria recognize dementia with Lewy bodies as a clinicopathological entity. Also recognized is the closely related (and perhaps biologically identical) entity of Parkinson's disease dementia, which differs from dementia with Lewy bodies only in the temporal sequence of appearance of clinical symptoms. The generic term "Lewy body disease" encompasses both entities. There is frequent and extensive overlap, both clinically and pathologically, between dementia with Lewy bodies and Alzheimer's disease. The two diseases share several genetic and environmental risk factors that have in common increased inflammatory states associated with increased disease risk. Moreover, pathological and experimental work has implicated the involvement of activated microglia and of microglia-derived interleukin-1 in the pathogenesis of the pathognomonic lesions of both diseases. Such neuroinflammatory processes may be the common link driving progression in both diseases and explaining the frequent overlap between the two diseases. PMID- 19300592 TI - Potential cognitive enhancing and disease modification effects of SSRIs for Alzheimer's disease. AB - OBJECTIVE: Selective serotonin reuptake inhibitors (SSRIs) have increased cognitive performance in some clinical studies of Alzheimer's disease (AD), but it is has been difficult to dissociate whether this is due to direct effects on cognition (neurochemical or disease-modifying) or a secondary effect of mood stabilization. We performed a systematic review for preclinical and human clinical trial evidence to support the use of SSRIs specifically for the management of cognitive decline in AD. DATA SOURCES: (1) PUBMED without language restrictions from 1950s until 2004 and updated August 2006, terms: "serotonin uptake inhibitors"[MeSH] AND ("Alzheimer disease"[MeSH] OR "Cognition Disorders"[MeSH]) NOT "Parkinson disease"[MeSH] AND (Clinical Trial[ptyp] OR Letter[ptyp] OR Meta-Analysis[ptyp] OR Randomized Controlled Trial[ptyp]) AND "alzheimer disease" [MESH] OR "Alzheimer*" combined with AND to "ssri*" OR "serotonin reuptake inhibitors" [MESH] NOT Review[ptyp]. (2) Cochrane Database of Systematic Reviews, keywords "SSRI" and "Alzheimer's". STUDY SELECTION: The PubMed search yielded 57 hits. Of these, 23 were included in this review for their specificity to SSRI use in AD or indications on efficacy beyond depressive symptoms. The other 34 citations were excluded because: (1) depression or other mood or behavioral disturbance severity was the reported outcome measure, (2) effects of SSRIs on cognition were confounded by concomitant use of other drugs, (3) subjects described were young adults, and/or (4) subjects had traumatic brain injury. The Cochrane Database of Systematic Reviews, 3rd Quarter 2006, yielded six citations related to SSRIs. DATA EXTRACTION: Data extracted from clinical trials included name of SSRI tested, cognitive outcome measures, and adverse events reported, which could include cognitive worsening. DATA SYNTHESIS: Preclinical evidence for use of SSRIs to enhance cognition in AD includes an effect at the hippocampus through carbonic anhydrase activation or stimulation of hippocampal neurogenesis. The chemical structure of paroxetine, and not intrinsic SSRI activity, may also affect APP ectodomain expression to reduce amyloid plaque formation. Clinical trials in AD generally have not assessed cognitive outcomes independently from mood or behavior stabilization. Currently, clinical studies in AD only indirectly support the use of SSRIs for disease modification by confirming a serotonergic deficit during the course of illness. CONCLUSIONS: Lack of supportive evidence for SSRIs as cognition enhancers or disease modifiers in AD is the result of omissions in clinical trial design, as opposed to reporting of negative outcomes. The preclinical evidence warrants the study of SSRIs in AD using mood, behavior, cognition, neurochemistry, and possibly neuroimaging as outcome variables. PMID- 19300593 TI - The clinical differentiation of fronto-temporal dementia from psychiatric disease. AB - OBJECTIVE: Frontal and/or temporal lobar atrophy (F/TA) is sometimes detected on neuroimaging in patients with psychiatric disease. This observation leads to difficulty in distinguishing whether patients have fronto-temporal dementia (FTD) or psychiatric illness. This paper sets out to develop clinical profiles that might be useful at first presentation to distinguish these two populations. METHODS: 29 patients were selected from a database of 250 current patients attending young onset dementia clinic. Control and experimental patient groups were established using DSM-5 criteria: (i) those without selective atrophy who had a psychiatric disorder (N = 5); (ii) patients who had FTD using consensus criteria (N = 13); and (iii) an experimental group of patients who had F/TA on neuroimaging, a psychiatric diagnosis and referral with possibility of a neurodegenerative disorder (N = 11). Profiles suggestive of FTD and psychiatric disease were established in the control groups utilising information from medical records, the neurological examination, the natural history and neuropsychometry to develop criteria to distinguish reliably FTD from psychiatric disease. These criteria were then applied to the experimental group. Patients were followed for five years. RESULTS: The developed criteria resulted in 3 patients being classified as FTD and 8 having psychiatric diagnoses in the experimental group. At follow-up, all the psychiatric patients remained functionally stable, whereas the FTD patients had deteriorated. CONCLUSION: Characteristic profiles may prove useful in the diagnosis of patients with F/TA on imaging with a psychiatric illness and help to distinguish them from patients with FTD. At first presentation F/TA has been found in some patients with psychiatric disease who do not develop evidence of neurodegeneration. This suggests that F/TA on neuroimaging might be a feature of a subgroup of patients with psychiatric diseases. PMID- 19300594 TI - Family history, early adversity and the hypothalamic-pituitary-adrenal (HPA) axis: Mediation of the vulnerability to mood disorders. AB - The effect of early-life vulnerability factors on the subsequent pathophysiology of severe mood disorders has yet to be fully elucidated. This study examines the relationship between early adverse life experience, family history and hypothalamic-pituitary-adrenal (HPA) axis function. Childhood trauma questionnaire (CTQ) scores, family history data and the cortisol response to the dexamethasone/corticotrophin releasing hormone (dex/CRH) test were examined in 40 patients with severe mood disorder. Normative data for the CTQ was also obtained. The study demonstrated that mood disorder patients reporting high levels of childhood emotional neglect (n = 26) had an HPA axis response which did not differ from controls, whereas patients reporting low levels (n = 19) had an enhanced response (p = 0.011). A positive family history of mood disorder further enhanced this response. These data suggest that early adverse life events and genetic susceptibility have dissociable effects on glucocorticoid receptor mediated negative feedback of the HPA axis in adult patients with severe mood disorders. PMID- 19300595 TI - Use of fMRI to predict psychiatric adverse effects of interferon treatment for Hepatitis C - preliminary report. AB - Interferon alfa2 (IFN-alpha2) is a parenterally administered cytokine used to treat patients with Hepatitis C and B, and malignancy. Interferon (INF) has a relatively high rate of central nervous system (CNS) adverse effects, including agitation, depression, fatigue, cognitive dysfunction, suicidal thought and drug craving. Using functional magnetic resonance imaging (fMRI) we studied patients with Hepatitis C virus (HCV) infection who were not more than mildly clinically depressed at baseline for their CNS reaction to IFN-alpha2. During fMRI, patients underwent visual stimulation with pictures designed to induce feelings of depression. In the two patients who became clinically depressed or markedly anxious while on treatment with interferon, but not in patients who did not experience these effects, there was a significant activation in specific areas of the brain known to be involved with depression, along with an increase above baseline in the Beck Depression Scale for the patient who developed INF-induced depression. The activation pattern differed from that previously observed for endogenous depression, indicating that INF-induced depression may differ in its underlying neuropathology. Functional magnetic resonance imaging can be an important tool in understanding and monitoring for (INF and other) medication induced CNS effects, and response to treatment. PMID- 19300596 TI - Structural integrity of the uncinate fasciculus in geriatric depression: Relationship with age of onset. AB - BACKGROUND: The uncinate fasciculus connects limbic structures, such as the hippocampus and amygdala, with frontal regions. This study utilized diffusion tensor imaging to examine the structural integrity of the uncinate fasciculus in late-life depression. METHOD: 18 elderly depressed and 19 elderly nondepressed subjects were matched for age and sex; 8 subjects had mid- to late-onset of depression while 10 subjects had early-onset depression. 3T diffusion tensor imaging-based fiber tract mapping delineated the uncinate fasciculus in each hemisphere, which guided measurement of the fractional anisotropy of the uncinate fasciculus in the temporal stem. After controlling for age and sex, differences between diagnostic groups were assessed. RESULTS: After controlling for age and sex, individuals with early onset depression exhibited lower anisotropy of the left uncinate fasciculus than did mid- and late-onset or nondepressed subjects (F(2,36) = 4.50, p = 0.02). Analyses of the right uncinate fasciculus were not statistically significant. CONCLUSIONS: This provides preliminary evidence that there is a structural connectivity deficit between left frontal and limbic structures in early-onset depression. Further work is needed to determine if this is seen in younger depressed subjects, and if it influences treatment outcomes. PMID- 19300597 TI - Episodic vertigo resulting from vascular risk factors, cervical spondylosis and head rotation: Two case reports. AB - BACKGROUND: Vascular risk factors predispose to vertebrobasilar ischemia. Cervical osteophytes can impinge on the vertebral artery causing mechanical occlusion during head turning. Presentation with vertigo in such instances is a common finding. CASE PRESENTATION: A patient with obesity, hyperlipidemia, hypertension, cervical spondylosis, and vertigo triggered by head rotation is presented. She responded to antihypertensive and lipid-lowering drugs, vestibular sedative and application of cervical collar. The second patient also exhibited similar features and responded to conservative treatment. DISCUSSION AND CONCLUSIONS: Rotational vertebral artery occlusion resulting from cervical spondylosis in the presence of atherosclerosed collateral vessels is a cause of posterior circulation insufficiency manifesting as vertigo. The tetrad of vertigo resulting from vascular risk factors, cervical spondylosis, and head rotation is proposed for further research. PMID- 19300598 TI - Carisoprodol withdrawal induced delirium: A case study. AB - A 43-year-old woman with chronic back pain found relief by taking carisoprodol, a centrally acting skeletal muscle relaxant. She had acquired large amounts of the prescription medication through the Internet and was taking approximately three hundred 350 mg tablets each week, at times up to fifty tablets per day. She then abruptly stopped the medication and presented to the emergency room one week later with waxing and waning attention, confusion, disorientation and visual hallucinations. Oral lorazepam was dosed according to a protocol employing the Clinical Institute Withdrawal Assessment of Alcohol Scale (CIWA). Her symptoms of delirium resolved rapidly and she was discharged home on day three. A review of the literature did not show any other reports of carisoprodol withdrawal induced delirium. Such withdrawal symptoms could be expected as the mechanism of action of carisoprodol is similar to that of hypnotic sedatives. Its availability and ease with which it can be acquired through the Internet puts it at great risk for a drug of abuse. PMID- 19300599 TI - Increase of plasma brain-derived neurotrophic factor levels in two psychotic depressed patients responding to lithium addition to paroxetine treatment. AB - We report two patients with psychotic depression who were successfully treated with a lithium addition to ongoing paroxetine treatment. In both cases, plasma brain-derived neurotrophic factor (BDNF) levels increased about 2-fold after lithium augmentation to paroxetine, compared with paroxetine treatment alone. Plasma paroxetine levels did not change after lithium addition. These results suggest that the increases in plasma BDNF levels reflect recovery from depressive symptoms in psychotic depression. PMID- 19300600 TI - Complex obsessive compulsive and impulsive symptoms in Tourette's syndrome. AB - Obsessive compulsive and impulsive symptoms are prevalent in patients with Tourette's Syndrome (TS) and well recognized as part of the TS spectrum. However, some complex obsessive compulsive and impulsive behaviors may be mistaken for other symptoms, such as complex tics or psychotic behavior. In addition, the overlap between tics, compulsions, and impulsive actions can make them impossible to discern from each other. Yet, the proper recognition of these symptoms is critical in optimizing treatment outcome in TS patients. This paper will review complex obsessive compulsive and impulsive behaviors that may occur in patients with TS and discuss implications for diagnosis and treatment. PMID- 19300601 TI - Gabapentin-induced coma: A MR-spectrometry analysis. AB - As for the majority of antiepileptic drugs, encephalopathy, manifested by transient somnolence, mood and motor disorders, is a possible side-effect. To our knowledge, there is little information about gabapentin-induced coma. We report a third case of gabapentin-induced coma where magnetic resonance-spectrometry was performed in diagnosis assessment. PMID- 19300602 TI - Effect of high-dose milnacipran in patients with depression. AB - To investigate the antidepressant effect of high-dose milnacipran, we retrospectively compared three groups of inpatients with major depression; those who were given milnacipran >100-150 mg/day (high-dose milnacipran group), those treated with milnacipran at maximum doses of 50-100 mg/day (standard-dose milnacipran group), and those treated with paroxetine at maximum doses of 40 mg/day (paroxetine group). The Hamilton Depression Rating Scale (HAM-D) scores of the three groups showed significant decrease at discharge compared to the scores at admission, indicating improvement of depressive symptoms for each group. However, the mean HAM-D score on admission was significantly lower for the standard-dose milnacipran group than the high-dose milnacipran and paroxetine groups. Additional intermediate assessment of the high-dose milnacipran group showed that the effect of milnacipran was dose-dependent with an additional improvement when patients were increase from 100 to 150 mg/day. These results suggest that patients suffering from moderate to severe depression with relative high HAM-D scores may benefit from treatment with high-dose milnacipran. PMID- 19300603 TI - Botulinum toxin - From WMD to therapeutic agent and cosmetic aid. PMID- 19300604 TI - Neuropsychological assessment, neuroimaging, and neuropsychiatric evaluation in pediatric and adult patients with sickle cell disease (SCD). AB - Traditionally, neuropsychological deficits due to Sickle Cell Disease (SCD) have been understudied in adults. We have begun to suspect, however, that symptomatic and asymptomatic Cerebrovascular Events (CVE) may account for an alarming number of deficits in this population. In the current brief review, we critically evaluated the pediatric and adult literatures on the neurocognitive effects of SCD. We highlighted the studies that have been published on this topic and posit that early detection of CVE via neurocognitive testing, neuropsychiatric evaluations, and neuroimaging may significantly reduce adult cognitive and functional morbidities. PMID- 19300605 TI - Role of endocannabinoids in regulating drug dependence. AB - This review will discuss the latest knowledge of how the endocannabinoid system might be involved in treating addiction to the most common illicit drugs. Experimental models are providing increasing evidence for the pharmacological management of endocannabinoid signaling not only to block the direct reinforcing effects of cannabis, opioids, nicotine and ethanol, but also for preventing relapse to the various drugs of abuse, including opioids, cocaine, nicotine, alcohol and metamphetamine. Preclinical and clinical studies suggest that the endocannabinoid system can be manipulated by the CB1 receptor antagonist SR141716A, that might constitute a new generation of compounds for treating addiction across different classes of abused drugs. PMID- 19300606 TI - Management of epilepsy in the elderly. AB - Epilepsy among the elderly is a frequently occurring pathology, differing in etiology, clinical presentation and prognosis from those of young people. In addition, beyond a certain age, physiological modifications are produced in the metabolism which alter the pharmacokinetics of antiepileptic drugs (AEDs), increasing the risk of pharmacological interactions, already greater in these patients due to the frequency of polypharmacy. Furthermore, elderly patients are particularly sensitive to certain secondary effects of AEDs, as for example, cognitive disturbances, osteoporosis or weight increase. Given that the efficacy of the major AEDs is a priori quite similar, and that the epilepsies occurring in this age-group generally have a good prognosis, the selection of an AED will depend more upon its pharmacokinetics and ability to induce certain secondary effects than on its efficacy. In this respect, levetiracetam and pregabalin, followed by oxcarbazepine and lamotrigine have the most favorable pharmocokinetical profile. Moreover, on the whole these drugs have very few cognitive effects, do not induce osteoporosis and, with the exception of pregabalin, do not affect weight, making them the first selection for use in the treatment of epilepsy in the elderly. PMID- 19300607 TI - Treatment of myoclonic seizures in patients with juvenile myoclonic epilepsy. AB - Drug treatment of Juvenile myoclonic epilepsy (JME) is mainly based on clinical experience and prospective and retrospective studies, with little evidence from randomized clinical trials. There are no head-to-head comparisons between old and new antiepileptic drugs (AEDs) and no drugs licensed specifically for JME. Valproate is unquestionably the drug of the first choice in men with JME. In women, lamotrigine should be preferred regarding teratogenicity and side effects of valproate. In addition, levetiracetam and topiramate are effective and can be use in combination or as second line treatment. Some AEDs can aggravate JME. In addition of AEDs, non-pharmacological treatments are important in JME. JME usually require lifelong treatment because seizures nearly always return after withdrawal of therapy. PMID- 19300608 TI - Sleep disturbances and suicide risk: A review of the literature. AB - A growing body of research indicates that sleep disturbances are associated with suicidal ideation and behaviors. This article (1) provides a critical review of the extant literature on sleep and suicidality and (2) addresses shared underlying neurobiological factors, biological and social zeitgebers, treatment implications, and future directions for research. Findings indicate that suicidal ideation and behaviors are closely associated with sleep complaints, and in some cases, this association exists above and beyond depression. Several cross sectional investigations indicate a unique association between nightmares and suicidal ideation, whereas the relationship between insomnia and suicidality requires further study. Underlying neurobiological factors may, in part, account for the relationship between sleep and suicide. Serotonergic neurotransmission appears to play a critical role in both sleep and suicide. Finally, it remains unclear whether or not sleep-oriented interventions may reduce risk for suicidal behaviors. Unlike other suicide risk factors, sleep complaints may be particularly amenable to treatment. As a warning sign, disturbances in sleep may thus be especially useful to research and may serve as an important clinical target for future suicide intervention efforts. PMID- 19300609 TI - A review on primary progressive aphasia. AB - Primary progressive aphasia (PPA) is a neurodegenerative disease of insidious onset presenting with progressive isolated loss of language function, without significant impairment in other cognitive domains. Current diagnostic criteria require the language dysfunction to remain isolated for at least two years, and to remain the salient feature as the disease progresses, usually to involve other domains such as behavior, executive functions, and judgment. Although PPA in its early stages can usually be differentiated from probable Alzheimer's disease (PRAD) and the behavioral variant of frontotemporal lobar degeneration by the absence of significant changes in memory and behavior, and the preservation of activities daily living, progression of the disease often leads to deficits more consistent with the latter. Underlying etiologies remain heterogeneous: the neuropathological characteristics associated with frontotemporal lobar degeneration, cortocobasal degeneration, and motor neuron disease are usually found. There is a strong genetic susceptibility with affliction of first-degree relatives with similar disease in up to 40 to 50% in some series. Pathogenic mutations in genes coding for the proteins tau and progranulin have been isolated. These are leading to a better understanding of the neuropathological mechanisms and hopefully targeted disease-modifying therapy. Current therapy is limited to improving mood symptoms and targeting behavior changes as they develop. Referral to specialized centers where speech therapy, counseling, and education for both patient and caregiver are available may be helpful. PMID- 19300610 TI - Adult neural stem cells: The promise of the future. AB - Stem cells are self-renewing undifferentiated cells that give rise to multiple types of specialized cells of the body. In the adult, stem cells are multipotents and contribute to homeostasis of the tissues and regeneration after injury. Until recently, it was believed that the adult brain was devoid of stem cells, hence unable to make new neurons and regenerate. With the recent evidences that neurogenesis occurs in the adult brain and neural stem cells (NSCs) reside in the adult central nervous system (CNS), the adult brain has the potential to regenerate and may be amenable to repair. The function(s) of NSCs in the adult CNS remains the source of intense research and debates. The promise of the future of adult NSCs is to redefine the functioning and physiopathology of the CNS, as well as to treat a broad range of CNS diseases and injuries. PMID- 19300611 TI - Review of botulinum toxin type A for the prophylactic treatment of chronic daily headache. AB - Botulinum toxin A is increasingly used in the treatment of idiopathic and symptomatic headache disorders. However, only few controlled trials are available and many trials can hardly be compared to each other because of different endpoints and different trial designs. In particular chronic daily headache, which is defined as an idiopathic headache occurring on more than 15 days per month for at least 3 months and a daily duration of at least 4 hours, is considered as a headache disorder with possible efficacy of botulinum toxin A. For the prophylactic treatment of chronic tension-type headache and chronic migraine, no sufficient positive evidence for a successful treatment can be obtained from randomized, double-blind, and placebo-controlled trials to date. For the treatment of chronic daily headache including medication overuse headache, there is some positive evidence for efficacy in a subgroup of patients. To date, the majority of double-blind and placebo-controlled studies do not suggest that botulinum toxin A is efficacious in the treatment of chronic idiopathic headache disorders. However, it is possible that some subgroups of patients with chronic daily headache will benefit from a long-term treatment with botulinum toxin A. PMID- 19300612 TI - Indiplon in the treatment of sleep disorders. AB - Indiplon is a novel sedative-hypnotic recently approved for the treatment of insomnia. Like other non-benzodiazepine hypnotics, its mechanism of action is to modulate subunits, especially the alpha-1 subunit, of the GABA receptor complex in order to induce sedation. Indiplon was developed in two different formulations to address two different types of insomnia complaint: indiplon-IR (immediate release) was designed for sleep onset difficulties, while indiplon-MR (modified release) was developed for sleep maintenance insomnia. While there are currently few peer reviewed articles about indiplon clinical trial results, the early information that is available seems to indicate that both formulations have been well tolerated and have proven effective at improving both patient reported and objectively measured sleep parameters in both adult and elderly insomnia patients. In May 2006, the FDA indicated that indiplon-IR was approvable and plans are to resubmit the application in 2007. Indiplon-MR was unapprovable and may require further evaluation. PMID- 19300613 TI - Rivastigmine for the treatment of dementia associated with Parkinson's disease. AB - Parkinson's disease (PD) afflicts millions of people worldwide and leads to cognitive impairment or dementia in the majority of patients over time. Parkinson's disease dementia (PDD) is characterized by deficits in attention, executive and visuospatial function, and memory. The clinical diagnostic criteria and neuropathology surrounding PDD remain controversial with evidence of overlap among PDD, dementia with Lewy bodies (DLB) and Alzheimer's disease (AD). Cortical cholinergic deficits are greater in PDD than in AD, and are well-correlated with the cognitive and neuropsychiatric dysfunction that occurs in PDD. Inhibition of acetylcholine metabolism is therefore a practical therapeutic strategy in PDD.This review examines current evidence for rivastigmine (a cholinesterase/butyrylcholinesterase inhibitor) treatment in PDD. In addition to its efficacy, we examine the safety profile, side effects, and cost effectiveness of rivastigmine in PDD. Rivastigmine provides modest benefit in PDD and further long-term studies are needed to determine the effectiveness and safety of rivastigmine over time. Tolerability is a problem for many PDD patients treated with rivastigmine. Future studies of rivastigmine in PDD should focus on pragmatic outcomes such as time to need for nursing home placement, pharmacoeconomic outcomes and simultaneous patient/caregiver quality of life assessments. PMID- 19300615 TI - Almotriptan in the treatment of migraine. AB - Almotriptan is an orally administered, highly selective serotonin 5-HT(1(B)/1(D)) receptor agonist that is effective in the acute treatment of moderate to severe migraine attacks. Since its introduction on to the market in 2001, several studies involving a large number of migraine patients have confirmed its efficacy and tolerability profile. Almotriptan, was found to be among the best-responding triptans in terms of pain relief and pain-free rate at 2 h. It has been reported that almotriptan has the best sustained pain-free (SPF) rate and the lowest adverse events (AEs) rate of all the triptans. When these clinical characteristics were combined to form the composite endpoint SPF and no AEs (SNAE), almotriptan emerged as the triptan with the best efficacy and tolerability profile. It also showed a good efficacy profile during the early treatment (within 1 h of onset) of migraine attacks characterized by moderate pain intensity. On the basis of these findings, almotriptan may be considered a therapeutic option for the acute treatment of migraine attacks. PMID- 19300614 TI - Neurologic uses of botulinum neurotoxin type A. AB - This article reviews the current and most neurologic uses of botulinum neurotoxin type A (BoNT-A), beginning with relevant historical data, neurochemical mechanism at the neuromuscular junction. Current commercial preparations of BoNT-A are reviewed, as are immunologic issues relating to secondary failure of BoNT-A therapy. Clinical uses are summarized with an emphasis on controlled clinical trials (as appropriate), including facial movement disorders, focal neck and limb dystonias, spasticity, hypersecretory syndromes, and pain. PMID- 19300616 TI - Topiramate in the treatment of partial and generalized epilepsy. AB - Topiramate (TPM) is a widely-used drug for the treatment of epilepsy. It is useful for several types of partial-onset and generalized-onset seizures, and is therefore considered a broad-spectrum agent. It is also effective as a prophylactic against migraine headaches. TPM was first approved for prescription use in 1996. In various countries it is now approved for adjunctive and monotherapy of partial-onset seizures and for therapy of generalized tonic-clonic seizures of nonfocal origin, for children and adults. For initial monotherapy of new-onset seizures, a target dose of 100 mg/day for adults is recommended. Adjunctive use with enzyme-inducing drugs and use for refractory seizures requires higher dosages, though the optimum dose for most patients does not exceed 400 mg/day. Excretion is primarily renal and TPM is not a significant hepatic enzyme inducer. Although it is usually safe and well-tolerated, adverse effects limit use in about 25% of patients. The most salient of these is cognitive dysfunction, especially problems with expressive speech and verbal memory. Weight loss, renal stones, paresthesias and other central nervous system side effects may occur. Tolerability is improved by low initial doses and slow titration to effect. PMID- 19300617 TI - Ziprasidone in the treatment of mania in bipolar disorder. AB - Ziprasidone is an atypical antipsychotic with a unique receptor-binding profile. Currently, ziprasidone is approved by the US Food and Drug Administration for the acute treatment of psychosis in schizophrenia and mania in bipolar disorder. When compared to certain other atypical antipsychotics, ziprasidone appears to have a relatively benign side effect profile, especially as regards metabolic effects eg, weight gain, serum lipid elevations and glucose dysregulation. Taken together, these data suggest that ziprasidone may be a first line treatment for patients with bipolar mania. However, ziprasidone is a relatively new medication for which adverse events after long-term use and/or in vulnerable patient populations must be studied. Unstudied areas of particular importance include the efficacy and safety of ziprasidone in the treatment of bipolar depression and relapse prevention of mania as, well as in the subpopulations of pregnant women, the elderly and pediatric patients. The emergence of mania in patients taking ziprasidone is another topic for further study. PMID- 19300618 TI - The therapeutic potential of escitalopram in the treatment of panic disorder. AB - Panic disorder is a chronic and disabling condition that is often accompanied by other psychiatric and medical conditions. The serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs) have been used effectively with panic disorder (PD) and conditions in which panic attacks frequently occur. Escitalopram is the most selective SSRI and a variety of evidence suggests it is of great value in the treatment of panic disorder. In this paper, we review the theoretical and practical implications of its use. PMID- 19300619 TI - Extended-release divalproex in bipolar and other psychiatric disorders: A comprehensive review. AB - Bipolar disorder can be a devastating disease state for individuals with the disease and also for family members. Proper recognition and treatment is vital to the successful management of this disease state. Through increased community and practitioner awareness, along with efforts to increase awareness for proper assessment, the rate of diagnosed bipolar disorder is increasing. Recent years have brought about the introduction of several new medications with approved indications for the treatment of bipolar disorder. In addition to new agents, traditional mood stabilizing medications have also been released in different formulations to better enhance tolerability without jeopardizing efficacy. One particular product is extended-release divalproex sodium. In the following article, we review the clinical presentation of bipolar disorder, its epidemiology, and the pharmacokinetics and mechanism of action for divalproex. In addition, we specifically review the role of extended-release divalproex in bipolar disorder through a critical analysis of the currently available published primary literature. PMID- 19300620 TI - Update on quetiapine in the treatment of bipolar disorder: results from the BOLDER studies. AB - The essential features of bipolar affective disorder involve the cyclical occurrence of high (manic or hypomanic episodes) and low mood states. Depressive episodes in both bipolar I and II disorder are more numerous and last for longer duration than either manic or hypomanic episodes. In addition depressive episodes are associated with higher morbidity and mortality. While multiple agents, including all 5 atypical antipsychotics, have demonstrated efficacy and earned US FDA indication for manic phase of bipolar illness, the acute treatment of bipolar depression is less well-studied. The first treatment approved by the US FDA for acute bipolar depression was the combination of the atypical antipsychotic olanzapine and the antidepressant fluoxetine. Recently, quetiapine monotherapy has demonstrated efficacy in the treatment of depressive episodes associated with both bipolar I and II disorder and has earned US FDA indication for the same. PMID- 19300621 TI - A review of quetiapine in combination with antidepressant therapy in patients with depression. AB - BACKGROUND: Atypical antipsychotics are increasingly used in the treatment of a broad spectrum of psychiatric disorders. There is evidence that in addition to treating the positive and negative symptoms of schizophrenia, as well as mania in bipolar disorder, these agents may have a potential role to play in the treatment of depressive disorders. In the following article we review the literature regarding the role of atypical antipsychotics, and specifically, quetiapine, in the treatment of major depressive disorder. MATERIALS AND METHODS: In March 2007 the authors performed a Medline search (English-language) using the keywords quetiapine and depression, revealing a total of 47 articles published. We also looked for cross-references in the published articles, obtained data-on-file from AstraZeneca Pharmaceutical L.P., and included abstracts presented at conferences and recent meetings. RESULTS: From our review we found that there is increasing literature supporting the efficacy of add-on quetiapine in the treatment of major depressive disorder. CONCLUSION: There is a need, however, for further well designed, adequately powered, randomized, controlled trials to confirm this finding, specifically in unipolar depression. PMID- 19300622 TI - Paliperidone ER: a review of the clinical trial data. AB - Paliperidone extended-release tablet (paliperidone ER; INVEGA()) is an oral antipsychotic for the treatment of schizophrenia. The recommended dose range is 3 12 mg per day. Paliperidone ER utilizes the OROS((R)) delivery system, which allows for once-daily dosing. Its pharmacokinetic profile results in a more stable serum concentration. Paliperidone is 9-hydroxyrisperidone, the chief active metabolite of risperidone. It undergoes limited hepatic metabolism, thereby minimizing the risks of hepatic drug-drug and drug-disease interactions. Three 6-week trials in patients with acute schizophrenia reported that paliperidone ER was effective, well tolerated, and produced clinically meaningful improvements in personal and social functioning compared with placebo. Post-hoc analysis of these trials in various populations, including recently diagnosed, elderly and more severely ill patients, those with sleep disturbances and those with predominant negative symptoms demonstrated improvement as well. Paliperidone ER was also significantly better than placebo in the prevention of symptom recurrence in a 6-month maintenance study. The most common clinically relevant adverse events associated with paliperidone ER were extrapyramidal symptoms, tachycardia and somnolence. The incidence of Parkinsonism, akathisia and use of anticholinergic medications increased in a dose-related manner. Further, modest QTc interval prolongation was observed but did not produce clinical symptoms. Similar to risperidone, paliperidone ER is associated with increases in serum prolactin levels. Overall, paliperidone ER was effective, well tolerated and provides a new treatment option for patients with schizophrenia. PMID- 19300623 TI - Clinical use of pregabalin in the management of central neuropathic pain. AB - Central neuropathic pain (central pain) is treated with antidepressants, various anticonvulsants, opioids, and cannabinoids, but in many cases treatment is insufficient and associated with a range of side-effects. This review addresses a new treatment for neuropathic pain, the anticonvulsant pregabalin. We review the pharmacology, mode of action, pharmacokinetics, and safety of pregabalin as well as two randomized efficacy studies in central pain and a brief overview of efficacy in peripheral neuropathic pain. Pregabalin appears to have efficacy in treating central pain comparable to that in peripheral neuropathic pain as well as efficacy of other recommended drugs for central pain. Pregabalin also improves disturbed sleep and anxiety. Pregabalin is well tolerated; the most common side effects are somnolence, dizziness, ataxia, and weight gain. Pregabalin is suitable for patients on multiple drugs although there may be additive CNS related side-effects. Thus, pregabalin has a primary role in central pain patients. PMID- 19300624 TI - What is the impact of physician communication and patient understanding in the management of headache? AB - Migraine is a common and debilitating condition. Despite the burden of disease and increasing availability of effective treatment, migraine management is unsatisfactory. Evidence in other chronic conditions indicates that effective physician communication results in better patient understanding and health outcomes.The current literature review was intended to evaluate evidence regarding the relationship of effective physician-provider communication to health outcomes and patient satisfaction among patients with migraine. The authors searched MEDLINE((R)) (1966-June 2007) and the Cochrane Database of Systematic Reviews for relevant publications. The search strategy combined the concepts of "headache disorders" and "physician-patient relations". 912 abstracts were identified, and 80 (9%) of them were included for data abstraction.There were no studies that met our eligibility criteria. Therefore we revised the eligibility criteria to allow for the inclusion of non-migraine primary headache disorders or the role of non-physician health care providers. Twelve published papers met the revised criteria. The findings from the limited evidence available suggests, but does not prove, that improvements in physician-patient communication could result in a significant decrease in the burden of suffering and health care resource utilization associated with migraine. More research is needed to assess the explicit role of physician-patient communication in the management of migraine. PMID- 19300625 TI - Depressive mixed state: Evidence for a new form of depressive state in type I and II bipolar patients. AB - BACKGROUND: A high proportion of unipolar and bipolar type II patients can present a depressive mixed state (DMX). This state is defined by an association of a major depressive episode with at least two specific hypomanic symptoms. This state seems underdiagnosed and this could have treatment implications. THE AIMS OF OUR STUDY WERE: (i) to investigate the frequency of DMX in type I and II bipolar patients hospitalized for a severe or resistant depressive episode and (ii) to assess the therapeutic response in naturalistic conditions. METHODS: Forty-two consecutive bipolar patients referred by psychiatrists for a severe or resistant depressive episode were assessed using the French version of the Mini International Neuropsychiatric Interview 5.0 (MINI 5.0), which assesses the suicide risk and provides DSM-IV diagnosis. The intensity of mood episodes was evaluated using the MADRS and Bech-Rafaelsen Mania Scale. One group of patients included patients presenting only depressive symptoms (ie, pure major depressive episode (MDE)), and the second group included patients with a major depressive episode and at least two specific hypomanic symptoms (DMX). RESULTS: Twenty-one patients (50%) had a pure MDE and 21 patients (50%) had a DMX. The treatment leading to recovery was very different in the two groups. Antidepressants were effective (77%) in MDE patients, whereas antipsychotics were effective (81%) in DMX. 38% of patients with a MDE also received a mood stabilizer versus 86% in the group of DMX. Five MDE patients (24%) and one DMX patient required electroconvulsive therapy. The suicidal ideations did not differ between the two groups (p = 0.7). CONCLUSIONS: Some mood episodes in bipolar patients (type I and II) are characterised by depressive and hypomanic symptoms but do not meet criteria for mixed episode as defined by DSM-IV. These episodes are often diagnosed as depressive states, but are worsened by antidepressants and often considered as resistant depression. They rapidly respond to antimanic treatment. New categories of mood disorders should take into account this particular mixed state. PMID- 19300626 TI - The patients' perspective: Results of a survey assessing knowledge about and attitudes toward depression in PD. AB - We report results of a survey assessing patients' knowledge about and attitudes towards depression in Parkinson's disease (PD). 345 patients from 8 tertiary care centers responded (43% response rate). Overall, patients were relatively knowledgeable about depression and its occurrence in PD. However, many patients believed that depression is a normal reaction to the illness. While many respondents would be reluctant to initiate a discussion of depression during a clinical evaluation, most would feel comfortable talking about depression with their physician if he or she asked them questions about their mood. Based on the results of this survey, we recommend the following approach for physicians: (1) inform PD patients that, although a frequent occurrence, depression need not be accepted as a "normal reaction" to PD; and (2) routinely inquire about depressive symptoms rather than waiting for the patient to spontaneously report them. PMID- 19300627 TI - Rule knowledge aids performance on spatial and object alternation tasks by alcoholic patients with and without Korsakoff's amnesia. AB - Delayed alternation (DA) and object alternation (OA) tasks traditionally have been used to measure defective response inhibition associated with dysfunction of frontal brain systems. However, these tasks are also sensitive to nonfrontal lesions, and cognitive processes such as the induction of rule-learning strategies also are needed in order to perform well on these tasks. Performance on DA and OA tasks was explored in 10 patients with alcohol-induced persisting amnestic disorder (Korsakoff's syndrome), 11 abstinent long-term alcoholics, and 13 healthy non-alcoholic controls under each of two rule provision conditions: Alternation Rule and Correction Rule. Results confirmed that rule knowledge is a crucial cognitive component for solving problems such as DA and OA, and therefore, that errors on these tasks are not due to defective response inhibition alone. Further, rule-induction strategies were helpful to Korsakoff patients, despite their poorer performance on the tasks. These results stress the role of multiple cognitive abilities in successful performance on rule induction tasks. Evidence that these cognitive abilities are served by diffusely distributed neural networks should be considered when interpreting behavioral impairments on these tasks. PMID- 19300628 TI - Repetitive transcranial magnetic stimulation for treatment of elderly patients with depression - an open label trial. AB - BACKGROUND: RTMS has been developed as a novel tool for treating depression but the clinical significance of this treatment has been variable, especially in the older depressed subjects. METHODS: Medication-resistant depressed patients 60 years or older were treated for two weeks (10 sessions) with high-frequency rTMS delivered to the left dorsolateral prefrontal cortex at 100% of motor threshold. Each session consisted of 20 trains at 10Hz delivered in 8-second duration. The patients continued taking their psychotropic medications throughout the study. RESULTS: Nineteen of the 20 subjects completed the trial. One subject dropped out after 8 sessions because of discomfort. The average age of our patients was 66.8 years (6 males and 14 females). Six patients responded and there was a 31.6% mean reduction in Hamilton Depression Rating Scale (HDRS) scores from baseline at the end of the treatment. There was statistically significant decrease from baseline in both HDRS and HARS scores at the end of treatment. rTMS was generally well tolerated. CONCLUSION: These preliminary finding suggests that rTMS may be an effective treatment alternative to a subpopulation of medication resistant older depressed patients. PMID- 19300629 TI - Catechol-O-methyltransferase Val158Met genotype in healthy and personality disorder individuals: Preliminary results from an examination of cognitive tests hypothetically differentially sensitive to dopamine functions. AB - A functional polymorphism of the gene coding for Catechol-O-methyltrasferase (COMT), an enzyme responsible for the degradation of the catecholamine dopamine (DA), epinephrine, and norepinephrine, is associated with cognitive deficits. However, previous studies have not examined the effects of COMT on context processing, as measured by the AX-CPT, a task hypothesized to be maximally relevant to DA function. 32 individuals who were either healthy, with schizotypal personality disorder, or non-cluster A, personality disorder (OPD) were genotyped at the COMT Val158Met locus. Met/Met (n = 6), Val/Met (n = 10), Val/Val (n = 16) individuals were administered a neuropsychological battery, including the AX-CPT and the N-back working memory test. For the AX-CPT, Met/Met demonstrated more AY errors (reflecting good maintenance of context) than the other genotypes, who showed equivalent error rates. Val/Val demonstrated disproportionately greater deterioration with increased task difficulty from 0-back to 1-back working memory demands as compared to Met/Met, while Val/Met did not differ from either genotypes. No differences were found on processing speed or verbal working memory. Both context processing and working memory appear related to COMT genotype and the AX-CPT and N-back may be most sensitive to the effects of COMT variation. PMID- 19300630 TI - Urban-rural differences in pediatric traumatic head injuries: A prospective nationwide study. AB - AIMS: To estimate differences in the incidence of recorded traumatic head injuries by gender, age, severity, and geographical area. METHODS: The study was prospective and nationwide. Data were collected from all hospitals, emergency units and healthcare centers in Iceland regarding all Icelandic children and adolescents 0-19 years old consecutively diagnosed with traumatic head injuries (N = 550) during a one-year period. RESULTS: Annual incidence of minimal, mild, moderate/severe, and fatal head injuries (ICD-9 850-854) was 6.41 per 1000, with 95% confidence interval (CI) 5.9, 7.0. Annual incidence of minimal head injuries (ICD-9 850) treated at emergency units was 4.65 (CI 4.2, 5.1) per 1000, mild head injuries admitted to hospital (ICD-9 850) was 1.50 (CI 1.3, 1.8) per 1000, and moderate/severe nonfatal injuries (ICD-9 851-854) was 0.21 (CI 0.1, 0.3) per 1000. Death rate was 0.05 (CI 0.0, 0.1) per 1000. Young children were at greater risk of sustaining minimal head injuries than older ones. Boys were at greater risk than girls were. In rural areas, incidence of recorded minimal head injuries was low. CONCLUSIONS: Use of nationwide estimate of the incidence of pediatric head injury shows important differences between urban and rural areas as well as between different age groups. PMID- 19300631 TI - Rivastigmine in Chinese patients with subcortical vascular dementia. AB - BACKGROUND: We explored the efficacy and tolerability of rivastigmine among Chinese patients with subcortical vascular dementia. METHODS: Forty subjects were randomized to either placebo (n = 20) or rivastigmine (n = 20) in a double-blind 26-week trial. Outcome measures were cognition (mini-mental state examination, frontal assessment battery), neuropsychiatric inventory (NPI), instrumental activities of daily living, clinical dementia rating scale, and adverse events. RESULTS: No statistical significant benefit could be observed in the active group in any of the efficacy measures. A trend favoring active group was observed only in the NPI subscore of irritability (p = 0.066) and aberrant motor behavior (p = 0.068). Withdrawal rate was 30% and 15% in the active and placebo group, respectively. CONCLUSION: Among Chinese subcortical vascular dementia patients, there was no apparent cognitive benefit associated with use of rivastigmine over the 6 months period. A trend favoring rivastigmine was observed in certain behavioral measures. Rivastigmine was associated with more withdrawals relative to placebo. PMID- 19300633 TI - Cognitive decline tracks motor progression and not disease duration in Parkinson patients. AB - We performed an analysis of prospectively-acquired cross sectional data on 106 Parkinson disease (PD) patients who underwent comprehensive neuropsychological testing and the Unified Parkinson Disease Rating Scale (UPDRS) motor scale. A significant correlation between the UPDRS motor and neuropsychological tests in all cognitive domains except for general intelligence and visuo-spatial function was seen. In this study, cognitive decline within this PD cohort correlated with motor impairment but not disease duration. Our findings suggest that overall cognitive impairment (except visuospatial dysfunction) may track motor progression in PD more than duration of disease. Longitudinal studies are needed to confirm our results. PMID- 19300632 TI - Effects of psycho-educational training and stimulant medication on visual perceptual skills in children with attention deficit hyperactivity disorder. AB - Attention deficit hyperactivity disorder (ADHD) is treated with stimulants and psycho-educational remedial programs despite limited literature support for the latter. This study aimed to examine changes in a "Test of Visual Perceptual Skills" (TVPS) that has not been previously reported in children with ADHD enrolled in such a program. METHODS: Sixteen children, 7-11 years old, with ADHD were involved in occupational therapy and special education geared towards attention training. Six months later methylphenidate 1 mg/kg/day was prescribed. It was not taken by eight children because of family choice. The TVPS was given twice, upon diagnosis, and 8 months post-intervention. The groups were compared by a repeated measures analysis of variance (ANOVA) with medication as a between groups factor and test-retest scores as within factor. RESULTS: All children demonstrated increases in total scores in the second measurement. Medicated children scored higher but ANOVA showed a nonsignificant F for the two groups, medicated and unmedicated (F = 0.0031, p = 0.9563), indicating a nondifferential effect of the two levels of treatment. It revealed a significant F for the pre- and post-treatment total TVPS scores (F = 30.91, p < 0.0001) indicating a significant difference between pre- and post-treatment tests. The interaction between pre-post treatment and level of treatment (medicated-unmedicated) was nonsignificant (F = 2.20, p = 0.1604). CONCLUSION: TVPS scores improved in all children following intervention. Medicated children did better, but differences were nonsignificant. PMID- 19300634 TI - Early rising children are more active than late risers. AB - BACKGROUND: A low level of physical activity impacts mental as well as physical health. This study investigated the daily lifestyle habits that affect physical activity in young children. METHODS: The relationship between physical activity, assessed by means of a Mini-Mitter Actiwatch device, and observed daily lifestyle habits was analyzed for 204 children, aged 12 to 40 months (average: 22.6 months), for whom 6-consecutive-day data from both the Actiwatch and sleep log were obtained. RESULTS: An older age, male gender, and early waking time showed significant positive correlations with physical activity level. Multiple regression analysis revealed that these three variables were significant predictors of physical activity. CONCLUSION: Promoting an early rising time is suggested to be an important element of cultivating good health in young children. PMID- 19300635 TI - Pharmacogenetics of antipsychotic adverse effects: Case studies and a literature review for clinicians. AB - There is a growing body of literature supporting the contribution of genetic variability to the mechanisms responsible for the adverse effects of antipsychotic medications particularly movement disorders and weight gain. Despite the current gap between research studies and the practical tools available to the clinician to identify such risks, it is hoped that in the foreseeable future, pharmacogenetics will become a critical aid to guide the development of personalized therapeutic regimes with fewer adverse effects. We provide a summary of two cases that are examples of using cytochrome P450 pharmacogenetics in an attempt to guide treatment in the context of recent literature concerning the role of pharmacogenetics in the manifestation of adverse effects of antipsychotic therapies. These examples and the review of recent literature on pharmacogenetics of antipsychotic adverse effects illustrate the potential for applying the principles of predictive, preventive, and personalized medicine to the therapy of psychotic disorders. PMID- 19300636 TI - Towards a cognitive stimulation program using an errorless learning paradigm in amnestic mild cognitive impairment. AB - Considering the high risk for amnestic mild cognitive impairment (A-MCI) individuals to progress towards dementia, it is crucial to study the efficacy of innovative treatment strategies such as cognitive stimulation techniques. The present study is a case report of two individuals presenting with A-MCI who were enrolled in a memory training program. After a broad neuropsychological assessment, the two participants were trained with an errorless (EL) learning paradigm on an individual basis, twice a week, over three weeks. Two follow-up sessions took place one and five weeks after the end of the training. Results showed that the program was well tolerated and feasible, and enhanced daily memory abilities. For the second participant only, a re-evaluation of her cognitive profile was completed 23 months after her first assessment and training. In addition, EL was directly compared with a control condition using an errorful (EF) learning paradigm to teach her new names over two sessions (one session for each condition). Her improvement on the trained material supported the preliminary efficacy of EL compared with EF for learning episodic material. These results are compatible with previous work that has preliminarily demonstrated the efficacy of an EL paradigm in patients with dementia. PMID- 19300637 TI - Improvement of advanced postvaccinal demyelinating encephalitis due to plasmapheresis. AB - We report a case of acute demyelinating encephalitis that occurred after viral vaccination against hepatitis A-, hepatitis B-, and poliovirus and vaccination against bacterial toxins of diphtheria and tetanus. After different diagnosis had been excluded, we diagnosed postvaccinal demyelinating encephalitis and started treatment with high dose intravenous methylprednisolone, followed by peroral application in decreasing dosages for three weeks. A few days after the treatment with methylprednisolone had been finished, the patient's medical condition deteriorated again. Thus, we initiated plasma exchange at an advanced state of illness, which led to significant continuous improvement. The role of plasma exchange is discussed controversially, in particular the issue of timing. We report a case that shows improvement due to plasmapheresis several weeks after symptom onset. PMID- 19300638 TI - Soil nematodes in terrestrial ecosystems. AB - There has been much work on plant-feeding nematodes, and less on other soil nematodes, their distribution, abundance, intrinsic properties, and interactions with biotic and abiotic factors. Seasonal variation in nematode fauna as a whole is correlated with factors such as moisture, temperature, and plant growth; at each site nematode distribution generally reflects root distribution. There is a positive correlation between average nematode abundance and primary production as controlled by moisture, temperature, nutrients, etc. Soil nematodes, whether bacterial feeders, fungivores, plant feeders, omnivores, or predators, all influence the populations of the organisms they feed on. Although soil trematodes probably contribute less than 1% to soil respiration they may play an important role in nutrient cycling in the soil through their influence on bacterial growth and plant nutrient availability. PMID- 19300639 TI - Influence of temperature on population development of eight species of pratylenchus on soybean. AB - In a soil temperature study, population increase on 'Clark 63' soybeatt was most rapid at 30 C in Pratylenchus alleni, P. brachyurus, P. cofleae, P. neglectus, P. scribneri, and P. zeae and at 25 C in P. penetrans and P. vulnus. The last two were the only species that reproduced at 15 C. Populations of all species increased over the range of 20-30 C, except those of P. neglectus at 20 C and P. coffeae, which was not tested below 25 C. Only P. brachyurus, P. neglectus, P. scribneri and P. zeae reproduced at 35 C. At their optimum temperatures, P. scribneri exhibited the greatest population increase, 1248-fold, and P. penetrans the least, 32-fold. This is the first report of soybean as a host for P. vulnus. PMID- 19300640 TI - Wave Forms of Caenorhabditis elegans in a Chemical Attractant and Repellent and in Thermal Gradients. AB - The wave forms and activity patterns of Caenorhabditis elegans were examined on agar in the presence of known chemical attractants (NaCl) and repellents (D tryptophan), and in thermal gradients. Total activity was reduced in both attractants and repellents. Different combinations of transfers between chemicals were investigated. Two thresholds were found for NaCl: 10(-3) M NaC1 caused reduced activity; 10 M NaCl increased reversals. D- or L-tryptophan influenced neither orientation nor the ability of thermally acclimatized individuals to remain at their eccritic temperature. PMID- 19300641 TI - Histochemical localization and nematoxicity of terpenoid aldehydes in cotton. AB - In healthy cotton, except for random occasional occurrence in cortical cells, terpenoid aldehydes (TA) are localized in the epidermis and, even there, are absent from the tip 2-4 cm of the root. Since constitutive TA do not occur in the endodermis and stele of the root, they cannot be effective agents against the development of the sedentary stage of the root-knot nematode, Meloidogyne incognita. Within 4 days after inoculation with the root-knot nematode, infection induced TA accumulated in the endodermis and outer stele. These induced TA were thus localized where they could be effective against the sedentary stage of the nematode. Infection-induced TA accumulation was more rapid and occurred in more stele cells in a resistant cotton cultivar than in two susceptible cultivars.TA extracts from cotton were inhibitory to nematode movement. All second-stage larvae exposed to 1,000 ppm TA for 3 h became rigid, made no movement, and appeared dead. Washing these larvae to remove the TA and incubating them for an additional 24 h did not change their appearance. Shorter exposure times or lower TA concentrations allowed some larvae to recover. Exposing larvae to 10 ppm of TA for 24 h had little effect on them. TA extracted from G. arboreum, a cotton that does not methylate TA, were slightly less inhibitory to the root-knot nematode than TA extracted front G. hirsutum which partially methylates TA. PMID- 19300642 TI - The Influence of Glomus mosseae on Tylenchulus semipenetrans-Infected and Uninfected Citrus limon Seedlings. AB - Greenhouse studies have shown that when rough lemon (Citrus limon) seedlings infected with TyIenchulus semipenetrans were transplanted into soil infested with Glomus mosseae, the mycorrhizal fungus infection increased seedling growth compared to nonntycorrhizal seedlings. Tylenchulus semipenetrans significantly suppressed seedling growth below that of mycorrhizal seedlings. Histological observations of nematode-free mycorrhizal roots showed that hyphae penetrated the epidermis and invaded the cortex, giving rise to arbuscules and vesicles. Nematode infection sites in T. semipenetrans-infected roots grown in soil infested with G. mosseae did not show evidence of vesicle development in the cortex but did show arbuscule development. PMID- 19300643 TI - Developmental nutrition of nematodes: the biochemical role of sterols, heme compounds, and lysosomal enzymes. AB - Attempts to develop defined in vitro culture systems for the growth, reproduction and development of free-living nematodes have yielded much basic information about their nutritional requirements and biochemistry. Requirements for sterol and heme have been identified suggesting that some nematodes lack de novo synthesis of these molecules. Possible pathways of metabolism of these nutritional requirements can be derived from in vitro experiments that use a variety of sterol and heine sources as supplements to the culture mediuin. These pathways are reviewed as well as the possible role of sterol and heme in the biology of free-living and parasitic nematodes, Since these molecules must be acquired dietarily, the possible involvement of lysosomal enzymes in digestion is discussed. Also considered is the possibility that lysosomal enzymes change when nematodes are fed on a heine protein source. PMID- 19300644 TI - Some factors causing variability in 1,2-dibromo-3-chloropropane concentrations in soil. AB - Relative to nematicides with greater fuming capabilities, 1,2-dibromo-3 chloropropane (DBCP) moved nonuniformly through soil. DBCP concentrations in soil were reduced by low soil temperature and the presence of lime or roots within the soil profile, Applications by either water or chisel injection provided DBCP movement to 120 cm and below. Concentrations were least persistent in the upper 15 cm of the field surface and in one situation where application was not followed by irrigation. Values for Henry's Constant are reported for DBCP at a range of solution temperatures. Certain advantages and disadvantages of soil atmosphere sampling of DBCP are discussed. PMID- 19300645 TI - Effects of Phenamiphos, Methyl Bromide, and Fallowing on Pratylenchus Penetrans, Yield of Medicago sativa, and Fusarium Infections. AB - A field study was made of the effects of a residual nematicide (phenamiphos), a fumigant (methyl bromide), and fallowing on the number of root lesion nematodes (Pratylenchus penetrans), forage yields of alfalfa, and the occurrence of Fusarium spp. in plant roots and soil. Fallowing controlled nematodes initially, but by the end of the second growing season, nematode numbers were as high as in plots which had grown a nematode-susceptible crop. Forage yield was greater in fallowed plots only for the first cut in the year after seeding. Fusarium in alfalfa roots and soil was not reduced by fallowing. Phenamiphos reduced nematode numbers, increased forage yields in 2 of 4 years, and reduced Fusarium infections of taproots. Soil fumigation with methyl bromide gave the best control of nematodes and Fusarium and gave significantly higher forage yields for the 4 years of study following fumigation. The 34% increase in alfalfa yield from fumigated plots over the 4 years indicates that the yield of alfalfa is being reduced significantly by microorganisms. The study does not establish the relative contributions of the root lesion nematodes and Fusarium spp. to the reduction. PMID- 19300646 TI - The Response of Citrus limon Seedlings to a Symbiont, Glomus etunicatus, and a Pathogen, Radopholus similis. AB - The influences of a vesicular-arbuscular mycorrhiza (Glomus etunicatus) and burrowing nematode (Radophohts similis), alone and in combination, on the growth of rough lemon (Citrus limon) seedlings were studied in the greenhouse. Growth of mycorrhizal seedlings was significantly greater than that of nonmycorrhizal seedlings or seedlings inoculated with R. sindlis. Mycorrhizal stimulation of seedling growth was inhibited by nematode infection. When seedlings were inoculated with G. etunicatus arid R. similis, suppression of seedling growth by R. similis was less on VAM seedlings than on nonmycorrhizal seedlings, Nonmycorrhizal seedlings infected with R. similis were significantly smaller than nonmycorrhizal seedlings free of R. similis. Vesicle formation and mycelia growth were less in nematode-infected roots. PMID- 19300647 TI - Screening soybeans for resistance to reniform nematode disease in the Philippines. AB - Sixty-five soybean varieties were tested in the field for resistance to Rotylenchulus reniformis. Criteria for resistance or susceptibility were root necrosis, nematode recovery from roots and soil, and egg production. Nine varieties were resistant, 13 moderately resistant, 26 moderately susceptible, and 17 susceptible. Linear correlations between resistance rating and each assessment parameter were highly significantly positive, suggesting that any of the parameters could be used to identify resistance. There were also highly significant positive linear correlations between any two combinations of parameters, indicating that they were reciprocally related. PMID- 19300648 TI - Mode of Parasitism of Meloidogyne and Other Nemaiode Eggs by Dactylella oviparasitica. AB - Hyphae of Dactylella oviparasitica proliferated rapidly through MeIoidogyne egg masses, and appressoria formed when they contacted eggs. The fungus probably penetrated egg shells mechanically, although chitinase production detected in culture suggested that enzymatic penetration was also possible. In soil, D. oviparasitica invaded egg masses soon after they were deposited on the root surface and eventually parasitized most of the first eggs laid. Occasionally the fungus grew into Meloidogyne females, halting egg production prematurely. The fungus parasitized eggs in the gelatinous matrix or eggs freed from the matrix and placed on agar or in soil. Specificity in nematode egg parasitism was not displayed, for D. oviparasitica parasitized eggs of four Meloidogyne spp., Acrobeloides sp., Heterodera schachtii, and Tylenchulus semipenetrans. In tests in a growth chamber, parasitism by D. oviparasitica suppressed galling on M. incognita-infected tomato plants. PMID- 19300649 TI - Interacting Effects of Soil Temperature and Type on Reproduction and Pathogenicity of Heterodera schachtii and Meloidogyne hapla on Sugarbeets. PMID- 19300650 TI - Pathogenicity of Macrophomina phaseoli on Jute in the Presence of Meloidogyne incognita and Hoplolaimus indicus. AB - Seedlings of Corchorus capsularis (cv. C4444) were inoculated with Meloidogyne incognita, Hoplolaimus indicus, and a fungus pathogen of jute, Macrophomina phaseoli, separately and in all possible combinations. The significant damage of jute plants caused individually by the pathogens was aggravated when the fungus was associated with either of the nematode species. M. incognita alone caused greater damage than either H. indicus or Macrophomina phaseoli alone. Plants inoculated with M. incognita and Macrophomina phaseoli were more severely damaged than plants inoculated with H. indicus and the fungus. Plant growth was minimum and disease symptoms were maximum when all pathogens acted together. In the presence of the fungus, M. incognita produced fewer galls. The reproduction of H. indicus was not influenced by the other organisms. PMID- 19300651 TI - Description and Bionomics of Mesomermis camdenensis n. sp. (Mermithidae), a Parasite of Black Flies (Simuliidae). AB - Mesomermis camdenensis n. sp. is described from larvae of Simulium tuberosum (lundstroem) collected in Camden Valley Creek, Washington County, New York. This species possesses a barrel-shaped vagina, vulval flap. two short separate spicules, terminal mouth, six longitudinal chords, six cephalic papillae, large sexually dimorpbic anaphids, an esophagns of uniform width which extends for less than one-third of the body length, and a cone-shaped tail directed ventrally without appendage. Juveniles also are described and illustrated.A detailed morphological comparison with the mermithid M. flumenalis Welch is presented. The most pronounced morphological differences between these species are in the shape of the vulva, juvenile tail, and infective stage. Cross-mating trials support the integrity of the new species.The life cycle of M. camdenensis is closely synchronized with that of its primary host, S. tuberosum larvae. Infected S. tuberosum larvae were first collected in May. Emergence of postparasites from late instars took place from mid-June through mid-October. Sampling data indicate a lower susceptibility to infection among S. venuslum Say larvae. PMID- 19300652 TI - Nonfumigant Nematicides for Control of Root-knot Nematode to Protect Carrot Root Growth in Organic Soils. AB - Greenhouse tests were conducted to determine the effects of two kinds of Meloidogyne hapla inoculum on the growth and quality of carrot roots, and the protection afforded in each case by nonfumigant nematicides in organic soils. For all treatments the percentage of carrots damaged was greater with larvae alone as inoculum than with larvae and eggs, indicating that most of the damage occurs early during formation of the taproot. Fosthietan, aldicarb, and oxamyl at 4 and 6 kg ai/ha protected the roots during formation and gave a lasting control of root-knot nematode. There was some nematode damage to the roots with phenamiphos and carbofuran at 4 and 6 kg ai/ha. Isazophos, diflubenzuron, and fenvalerate gave little protection to carrot roots and did not control root-knot nematode effectively. PMID- 19300653 TI - Helicotylenchus stylocercus n. sp. and Rotylenchus phaliurus n. sp. (Nematoda: Hoplolaimidae) from Costa Rica. AB - Two new species of plant-parasitic nematodes from Costa Rica are described. Helicotygenchus styloeercus n. sp., from soil around roots of banana at Coto, is distinguished hy the female tail, which bears a large pillarlike ventral projection. Rotylenchus phaliurus n. sp., from soil artmnd roots of Dioscoroea sp. at Sixaola, differs from R. caudaphasmidius in having the conus equal to or more than half the spear length, and large terminal annules on the female tail. PMID- 19300654 TI - Histological Study of the Compatible and Incompatible Interaction of Soybeans and Meloidogyne incognita. PMID- 19300655 TI - New species of bunonematoidea (rhabditida) from georgia and tennessee. AB - Three new species of Bunonematidae (Bunonema husseyi, Rhodolaimus dimorphus, R. stephaniae) and one of Plerygorhabditidae (Pterygorhabditis panoplus) are described from Georgia and Tennessee. The juvenile external morphology of P. panoplus is described and illustrated. A lectotype of P. pakistanensis is designated and the two species compared, and the dissimilar nature of cuticular tubercles in Bunonema and Rhodolaimus is discussed. PMID- 19300656 TI - Partial Characterization of the Cuticle Surface of Meloidogyne javanica Females. AB - Negative charges on the outer cuticular surface of Meloidogyne javanica females were visualized with electron microscope labelling techniques. Evidence is presented that the electronegative charge is not borne on neuraminic acid. Ruthenium red staining indicated acid mucopolysaccharides on the outer surface. A surface coat, or glycocalyx, external to the outer cuticle membrane was demonstrated. PMID- 19300657 TI - Heterodera canadensis n. sp. (Nematoda: Heteroderidae) from Spike-Rush (Eleocharis acicularis (L.) R. &S.) in Quebec, Canada. AB - Heterodera canadensis n. sp. is described and illustrated from the roots of spike rush, Eleocharis acicularis (L.) R. &S., in Deschenes, Quebec. This new abullate species is related to Heterodera graminophila Golden and Birchfield, 1972, but differs significantly in cyst shape, cone top structures, body length of the second-stage larva (520-600 mum, vs. 380-400 for H. graminophila) and tail length (110-120 mum, vs. 57-67 for H. graminophila). A taxonomic key based on cyst and second-stage larva characters is provided for identification of the fifteen species in the Heterodera goettingiana group. PMID- 19300658 TI - Polyploidy in an Amphimictic Population of Heterodera glycines. AB - A tetraploid single-cyst isolate of Heterodera glycines from a field population from Indiana has been propagated in the greenhouse on Lee soybeans since its discovery, in 1973. The tetraploid isolate has n = 18 chromosomes, compared with n = 9 of the diploid H. glycines; it has larger cysts and larvae, but shows the same level of parasitism and host range as the diploid population from which it apparently evolved. Association of chromosomes is irregular at metaphase I, with quadrivalents, trivalents, and univalents often observed in addition to the bivalents. The second maturation division is usually normal. About 80% of the mature oocytes (just before fertilization) have n = 18, and the other 20% have n = 17 or 19. Reproduction of the tetraploid isolate is exclusively by cross fertilization. The discovery of such a tetraploid provides an experimental tool for the study of polyploidy in nematodes. Many amphimictic plant-parasitic nematodes are suspected of representing polyploids. PMID- 19300659 TI - Response of Pinus ponderosa Seedlings to Stylet-Bearing Nematodes. AB - Of 12 stylet-bearing nematodes used for inoculations, Pratylenchus penetrans, P. brachyurus, P. vulnus, Ditylenchus destructor, Meloidogyne incognita, M. javanica, and M. hapla reproduced on Pinus ponderosa, while Xiphinema index, Aphelenchus avenae, Paratylenehus neoamblycephalus, Tylenchulus semipenetrans, and Macroposthonia xenoplax did not. P. vulnus, P. brachyurus, P. penetrans, A. avenae, D. destructor, T. semipenetrans, and P. neoamblycephalus significantly suppressed both the shoot and root wet weights of ponderosa pine seedlings obtained from stands in five different locations. X. index significantly suppressed root wet weights, M. xenoplax siguificantly suppressed shoot wet weight, and M. incognita, M. javanica, and M. hapla suppressed neither at the inoculation levels used. Injurious nematodes tended to suppress root growth more than shoot growth. Seedlings from two locations produced greater shoot growth wet weight than did seedlings from the other three locations. The more injurious nematodes tended to cause an increase in the water content of shoots. Frequency analyses of seedling population shoot-root ratios indicated that ponderosa pine seedlings could be selected for better shoot-root ratios as well as for resistance to several pathogenic nematodes. PMID- 19300660 TI - Control of Tylenchulus semipenetrans on Citrus With Aldicarb, Oxamyl, and DBCP. AB - Soil application of DBCP (l,2-dibromo-3-chloropropane) and foliar applications of oxamyl (methyl N',N'-dimethyl-N-[(methylcarbamoyl)oxy]-l-thiooxamimidate) were compared for control of Tylenchulus semipenetrans in a grapefruit (Citrus paradisi) orchard, DBCP reduced nematode populations and increased fruit growth rate, fruit size at harvest, and yield compared to the untreated controls in the 2 years following treatments. Foliar applications of oxamyl reduced nematode populations and increased fruit growth rate slightly the first year, but not in the second. Foliar applications of oxamyl did not improve control attained by DBCP alone. Soil application of aldicarb [2-methyl-2-(methylthio)propionaldehyde O-(methylcarbamoyl)oxime] or DBCP to an orange (C. sinensis) orchard reduced T. semipenetrans populations in the 3 years tested and increased yield in 1 of 3 years. Aldicarb treatment reduced fruit damage caused by the citrus rust mite, Phyllocoptruta oleivora. Aldicarb, applied at 5.7 or 11.4 kg/ha, by disk incorporation or chisel injection, was equally effective in controlling nematodes, improving yields, fruit size, and external quality. In a grapefruit orchard, chisel-applied aldicarb reduced nematode populations and rust mite damage and increased yields in both years and increased fruit size in one year. The 11.4-kg/ha rate was slightly more effective than the 5.7-kg/ha rate. Aldicarb appears to be an adequate substitute for DBCP for nematode control in Texas citrus orchards and well-suited to an overall pest management system for Texas citrus. PMID- 19300661 TI - Histological Responses of Four Leguminous Crops Infected with Meloidogyne incognita. AB - Histological responses to Meloidogyne incognita infection in Rhizobium nodules of clover, horsebean, lupine, and pea were investigated. The formation of giant cells in vascular bundles of nodules and roots, and the basal connection of the nodule, were usually associated with abnormal xylem and/or deformed xylem strands. However, giant cells did not disturb or prevent the development of nodular tissues. Areas in which galls formed, wall thickness of giant cells, and number of giant cells around the nematode head varied with plant species. Ranking by gall size and giant-cell wall thickness was horsebean > lupine and pea > clover. The multinucleate condition in giant cells resulted from repeated mitoses without subsequent cytokinesis. The resulting nuclei agglomerated in irregularly shaped masses in some giant cells. PMID- 19300662 TI - Vegetable oils as protectants against nematode infections. PMID- 19300663 TI - Population response to temperature in the subfamily tylenchorhynchinae. AB - The effects of temperature on population development of 11 species of stunt nematodes in the subfamily Tylenchorhynchinae were compared on red clover or Kentucky bluegrass in constant-temperature tanks at 5-degree intervals from 10 to 35 C. The optimum temperature for population increase on red clover in 90 days was 30 C for Tylenchorhynchus agri, T. nudus, T. martini, and T. clarus, 25 C for T. sylvaticus and T. dubius, and 20 C for T. canalis, Merlinius brevidens, and Quinisulcius capitatus. The optimum was 30 C for T. robustoides and 25 C for T. maximus on Kentucky bluegrass. The temperature range for population increase was 20-35 C for T. agri, T. nudus, T. martini, and T. clarus, 20-30 C for T. sylvaticus and T. robustoides, 15-25 C for T. maximus, 10-25 C for T. dubius, and 10-20 C for M. brevidens and Q. capitatus. T. canalis increased only at 20 C. All species were recovered in numbers near their inoculum level at 10 C. There was no survival of T. sylvaticus, T. dubius, T. canalis, T. robustoides, T. maximus, M. brevidens, and Q. capitatus at 35 C, or of the last three of these species at 30 C. Temperature had no effect on sex ratio in final populations. Population increase was greatest in T. martini and least in T. canalis. PMID- 19300664 TI - Managing nematode population densities on tomato transplants using crop rotation and a nematicide. AB - Millet, milo, soybean, crotalaria, and Norman pigeon pea were used in conjunction with clean fallow and a nematicide (fensulfothion) for managing nematode populations in the production of tomato transplants (Lycopersicon esculentum Mill.). Glean fallow was the most effective treatment in suppressing nematode numbers. After 2 years in tomato, root-knot nematodes increased in numbers to damaging levels, and fallow was no longer effective for complete control even in conjunction with fensulfothion. After 4 years in tomato, none of the crops used as summer cover crops alone or in conjunction with fensulfothion reduced numbers of root-knot nematodes in harvested tomato transplants sufficiently to meet Georgia certification regulations. Milo supported large numbers of Macroposthonia ornata and Pratylenchus spp. and crotalaria supported large numbers of Pratylenchus spp. Millet, milo, soybean, crotalaria, and pigeon pea are poor choices for summer cover crops in sites used to produce tomato transplants, because they support large populations of root-knot and other potentially destructive nematodes. PMID- 19300665 TI - Resistance of Anhydrobiotic Aphelenchus avenae to Methyl Bromide Fumigation. AB - The effect of methyl bromide (MB) was tested on active and anhydrobiotic Aphelenchus avenae. A. avenae was induced into anhydrobiosis by three different techniques. Both active and anhydrobiotic nematodes were subjected to 3,000 mu1 MB/liter air for 14 periods from 0 to 82 h. Anhydrobiotic nematodes were more resistant to fumigation than active nematodes, regardless of the technique used to induce anhydrobiosis. The percent survival decreased with increasing MB exposures (mu1 MB x h). For an LD of 45,000-54,000 mu1/1 x h were required for active nematodes and >279,000 mu1/1 x h for anhydrobiotic nematodes. PMID- 19300666 TI - Morphological comparison of meloidogyne males by scanning electron microscopy. AB - Males of five populations of Meloidogyne hapla were compared by scanning electron microscopy (SEM). Three populations of race A had haploid chromosome numbers of 15, 16, and 17 and reproduced by facultative parthenogenesis. Race B consisted of two mitotically parthenogenetic populations with somatic chromosome numbers of 45 and 48. Males of one population each of M. arenaria, M. incognita, and M. javanica were also examined to delineate species differences. The populations of M. arenaria, M. incognita, and M. javanica had 54, 41-43, and 44 chromosomes, respectively, and reproduction was by mitotic parthenogenesis. Observations were made on head structures, lateral field, excretory pore, and tail. The expression of labial and cephalic sensilla, shape and proportion of labial disc and lips, and markings on the head region were distinctly different for each species. The head morphology of the two cytological races of M. hapla was dissimilar. Populations of race A were different from each other and showed intrapopulation variation. Populations of race B were morphologically similar and stable in head morphology. The structure of the lateral field, excretory pore, and tail was of little value in distinguishing species or populations because of inter- and intrapopulation variation. The results are discussed in relation to earlier SEM observations of second-stage juveniles of the same populations. PMID- 19300667 TI - Morphological comparison of meloidogyne males by scanning electron microscopy. AB - Field plots of Tifton loamy sand were treated with methyl bromide, DD-MENCS, or ethoprop for control of root-knot nematodes, Meloidogyne incognita, in a multiple cropping system of turnips, field corn, and southern peas. Annual applications of methyl bromide and DD-MENCS in November or December suppressed nematode numbers to very low levels through September, but numbers increased in the following October, November, and/or December. No benefit was found from ethoprop applied to DD-MENCS-treated plots before the planting of each crop. Nematode numbers were not significantly suppressed by ethoprop alone. Concentrations of ethoprop in the 0-15-cm soil layer were near 6 mug/g at application but were < 1mug/g of soil 5 days later on corn and southern peas and 30 days later on turnips. Ethoprop concentrations of 4.6 to 5.6 mug/g of soil are too low for adequate control of root-knot nematodes on field corn and southern peas in multiple cropping systems. Stepwise regression analyses indicated that 81% and 36% of the variations in concentration of ethoprop in the soil were attributable to the amount of water that the plots received when the maximum soil temperature ranged from 10 C to 19 C and from 31 C to 41 C, respectively, and that 11% was attributable to the maximum soil temperature within the temperature range of 17 C to 33 C. PMID- 19300668 TI - Dispersion, Dissipation, and Efficacy of Methyl Bromide-Chloropicrin Gas vs. Gel Formulations on Nematodes and Weeds in Tifton Sandy Loam. AB - Dispersion, dissipation, and efficacy of gas and gel formulations of methyl bromidechloropicrin (202, 269, 336, and 403 kg/ha) on nematodes and weeds on tomato were studied in field plots. Concentrations of methyl bromide and chloropicrin 4 hr after soil treatment were greater at a depth of 15 cm than at 30, 45, or 60 cm. The concentrations of both chemicals decreased with lower doses, greater depths, and longer times after application. The gel formulation was more persistent than the gas formulation at both 336 and 403 kg/ha at depths of 30 and 45 cm, especially 24 and 36 hr after chemical application. Plant growth and yield were improved when nematodes and weeds were controlled. PMID- 19300669 TI - Mass Production of the Mosquito Parasite Romanomermis culicivorax: Effect of Density. AB - When numbers of Romanomermis culicivorax Ross and Smith were varied in containers with a constant surface area and depth of sand, densities of 12-24 nematodes per cm(2) yielded significantly more preparasites than higher densities. When container surface area and numbers of nematodes were constant and sand volume and depth were varied, yields did not differ significantly. When numbers of nematodes and sand volume were constant and surface area and sand depths were varied, yields were significantly higher for a density of 24 nematodes per cm(2). Yields of preparasites were tripled by simply setting up three cultures, each containing 5 g of nematodes, instead of a single culture containing 15 g. PMID- 19300670 TI - Feeding Behaviour and Histopathology of Hirschmanniella oryzae, H. imamuri, and H. spinicaudata on Rice. AB - The feeding behaviour of Hirschmanniella oryzae, H. imamuri, and H. spinicaudata on excised rice roots was very similar to that of other tylenchid nematodes on other hosts. Juveniles and adults of all three species invaded rice roots, causing extensive mechanical damage. Feeding and burrowing activities predisposed roots to secondary infection in unsterile cultures. Surface-sterilized nematodes did not induce necrosis in axenic cultures. PMID- 19300671 TI - Effect of Nonhost Cultivars on Heterodera schachtii Population Dynamics. AB - Broadcast plantings of nonhost cultivars (alfalfa, barley, bean, onion, potato, and wheat) in soil in redwood boxes (4.2 x 30 x 14 cm) infested with Heterodera schachtii reduced the initial nematode populations (P = 0.05). The reduction was greater with sugarbeets, a host, than with all other cropping treatments except onion, bean, and fallow (P = 0.05). After 80 days, when the root growth of all treatments had completely penetrated the soil, the nematode population was lower under onion than under wheat and barley (P = 0.05). The terminal nematode population (160 days) was lowest under onion, followed by bean, potato, fallow, and alfalfa. The nematode population was less under onion than under fallow, alfalfa, barley, and wheat (P = 0.05). Bean, potato, and fallow nematode populations were less than barley populations (P = 0.05). When broadcast plantings of these cultivars were simulated in microplots, the terminal population (100 days) was significantly lower under onion and bean than fallow (P = 0.05). However, no significant differences in reduction of H. schachtii population density were obtained when commercial row plantings of these crops were simulated in microplots. H. schachtii suppressed growth of barley, tomato, and sugarbeet, but not of bean, onion, alfalfa, or wheat in the greenhouse. Only the growth of sugarbeet was suppressed significantly in the field (P = 0.05). PMID- 19300672 TI - Interrelations between Meloidogyne javanica, Rotylenchulus reniformis, and Rhizobium Sp. on Vigna sinensis. AB - The interactions of Meloidogyne javanica, Rotylenchulus reniformis, and Rhizobium sp. on cowpea seedlings were investigated. Upon simultaneous inoculation with the two nematode species, M. javanica invaded first but did not affect root invasion by R. reniformis. M. javanica populations increased less in competition with R. reniformis than when present alone. Preinvasion by R. renilormis significantly suppressed the number of M. javanica in the roots. Inoculation of M. javanica and/or R. reniformis with rhizobia did not affect nodulation. Nodule formation was hindered only when R. reniformis infection preceded rhizobial inoculation. Nitrogen nodules were formed on M. javanica galls. Nodules and M. javanica galls served as infection sites for both nematodes. Although R. reniIormis on the roots reduced the space for M. javanica infection, M. javanica is more competitive than R. reniformis and ultimately predominates as a result of its higher reproductive potential and shorter time spent in the soil before infection. PMID- 19300673 TI - Biology, Life Cycle and Redescription of, Neoaplectana bibionis Bovien, 1937 (Nematoda: Steinernematidae). AB - A greater knowledge of the morphology and the development of the infective third larval stage of Neoaplectana bibionis Bovien, 1937, a parasite of arthropods, makes it possible to recognise the second larval stage and present a description of all the developmental stages in the life cycle of the species. Four larval stages can be recognised. L1 hatches from the egg. In a suitable host, when the population density is low, it develops directly into L4. When the population density is high, it develops into L2. The L2 is a non-feeding stage that precedes the resistant infective L3 larvae. L3 is free-living and characterised by a distinct lateral field with nine longitudinal lines. In the L4 the sex can be recognised. The first adults developing in a fresh host are usually larger than those developing later. Generally no more than two generations are completed in one insect. Youngg females in a fresh host lay eggs. In older females, and in females developing in a decaying host, endotokia matricida is common. A redescription of the species is presented on the basis of a population from New Zealand reared on Galleria mellonella larvae. All stages of the life cycle are described and illustrated. Proposed new synonymy includes Neoaplectana leueaniae Hoy, 1954 a synonym of N. bibionis Bovien, 1937 and Neoapleetana affinis Bovien, 1937 a synonym of N. menozzi Travassos, 1932. PMID- 19300674 TI - Electron Microscopy of the Stomatostylet and Esophagus of Criconemoides curvatum. AB - The stomatostylet of Criconemoides curvature consists of three parts: tooth cone, shaft, and knobs. The tooth cone constitutes the outer conical covering and inner lining of the anterior half of the stylet lumen. The tooth cone is easily separated from the shaft by treating an isolated styler with 0.5% sodium hypochlorite. The posterior half of the shaft is cylindrical, tapering anteriorly to form the shaft extension, wedged between the inner and outer tooth cone. The shaft extension extends to the stylet lumen orifice, which is subterminal and ventral. Six ducts enter the shaft through the junction between the shaft and knobs. They extend anteriad toward the tip of the shaft extension. Cytoplasmic connections between the ducts and the cells surrounding the stylet occur near tile junction between the shaft and the basal knobs. Ribosome and membranous structures are observed in these ducts. The esophagus of the adult female consists of a fused procorpus and metacorpus with a large valve possessing thickened cuticular walls at the anterior and posterior ends. The dorsal esophageal gland reservoir is composed of many honeycomb-like compartments made up of two types of differing electron density. The subventral esophageal glands, however, consist of only one type of granules. Both dorsal and subventral esophageal glands open into the esophageal lumen through trachea-like branched multiple canals. PMID- 19300675 TI - Histopathology of Rotylenchulus reniformis on Sunflower. PMID- 19300676 TI - Reproduction and Pathogenicity of Three Isolates of Meloidogyne hapla Race A on Concord Grapes. PMID- 19300677 TI - Embryogenesis of Hemicycliophora epicharoides Loof, 1968 (Nematoda: Hemicycliophorinae), and Description of the Male. PMID- 19300678 TI - Effect of Phenamiphos on Heterodera schachtii and Meloidogyne javanica. AB - Aqueous solutions of technical-grade phenamiphos [ethyl 3-methyl-4-(methylthio) phenyl (1-methylethyl) phosphoratnidale] were used in hatching chambers to test, under laboratory tory conditions, the effect of phenamiphos on the hatching and movement of Meloiclogyne javanica and Heterodera schachtii. Hatch of M. javanica and H. schachtii eggs was depressed 70 and 88% by nematicide at 0.48 and 4.80 mug/ml, respectively. The infectivity of second-stage larvae of both species was affected by concentrations as low as 0.01 mug/ml. At least 0.5 mug/ml was required to decrease the movement of larvae of M. javanica and H. schachtii. To decrease the movement of H. schachtii males toward females, 10 mug/ml was required. In a field experiment using a 15% granular formulation, 5 kg/ha a.i. significantly reduced infection of sugarbeet roots by H. schachtii. PMID- 19300679 TI - The Biology of Rotylenchulus macrodoratus. AB - Rotylenchulus macrodoratus completed its embryogenic development in about 16-19 days at 18-32 C. On olive seedlings the life-cycle from egg to egg was completed in 45-55 days. The first molt occurred in the egg while the other three were superimposed, with retention of successive larval cuticles. Only immature vermiform and swollen egg-laying females were found attached to olive roots. Eggs are laid in a gelatinous matrix on the root surface. The maximum number of eggs seen was 55. Males were not parasitic. Dianthus barbatus, Parietaria officinalis, and Eriobotrya japonica were found to be hosts of the parasite. Observed in all infested hosts was an uninucleate giant cell that expanded from the endodermis toward the center of the stele in primary roots, and from the secondary vascular tissue toward the periphery of the cortex in secondary roots. PMID- 19300680 TI - Behavioral Effects of Carbofuran and Phenamiphos on Pratylenchus vulnus. I. Motility and Dispersion. AB - The motility and dispersion of Pratylenchus vulnus was affected by lower concentrations of phenanliphos (an organophosphate) than of carbofuran (a carbamate). At the higher concentrations in the active ranges, somatic musculature activity was inhibited while the activity of internal-organ muscnlature persisted. Treated nematodes recovered when placed in aerated distilled water, but recovery decreased as treatment concentrations increased. Second-stage larvae dispersed more slowly than later stages or adults. Dispersion from a point source followed a natural log pattern under ideal conditions, similar to a diffusion pattern. PMID- 19300681 TI - Behavioral Effects of Carbofuran and Phenamiphos on Pratylenchus vulnus. II. Attraction to Bean Roots. AB - Pratylenchus vulnus (L, L, adults) are attracted to the roots of growing bean seedlings. The attraction is inhibited by treating the nematodes with solutions of carbofuran or phenamiphos at concentrations below those necessary to inhibit motility and dispersion. Nematicide treatments of plants without treatment of nematodes are ineffectual because nematodes are attracted equally well to treated and untreated seedlings. PMID- 19300682 TI - Behavioral Effects of Carbofuran and Phenamiphos on Pratylenchus vulnus. III. Penetration and Development. AB - Penetration of bean roots by Pratylenchus vulnus was inhibited by continuous exposure of the nematode to carbofuran and phenamiphos and by drenches of higher concentrations of these chemicals. The inhibition was explicable by inhibition of motility, dispersion, and attraction. If incubated in aerated distilled water, nematodes treated with carbofuran and phenamiphos recovered and reproduced as well as untreated nematodes. Foliar treatments were ineffective. Apparently, no basipetal transport of carbofuran and phenamiphos occurs in beans. Both nematicides arrested nematode development by interfering with egg production and transitions between life stages. PMID- 19300683 TI - "Blinding" of shoots and a leaf gall in Amsinckia intermedia induced by Anguina amsinckia (Steiner and Scott, 1934) (Nemata, Tylenchidae), with a note on the absence of a rachis in A. amsinckia. AB - "Blinding" and a leaf gall induced on Amsinckia intermedia Fisch. and Mey. by Anguina amsinckia (Steiner and Scott) are described. A. amsinckia induced blinding by galling the terminal apical meristem of its host. The leaf gall was formed by a ventral curling of the distal edge of the leaf. The absence of a rachis in the ovary of A. amsinckia is noted. PMID- 19300684 TI - Comparative calorie values of nematodes. AB - Calorie values for a wide biological selection of nematodes, determined with a microbomb calorimeter, ranged from 3.86 to 6.85 Kcal/g. The mean of 5.095 Kcal from 16 species was lower than means recorded in three previous studies of other invertebrate groups. The nematode values were skewed to the lowest limit. Larvae of Ditylenchus dipsaci showed lower calorie values after storage, and the calorie values of separate tissues of Ascaris lumbricoides were highest for eggs and the intestine and lowest for cuticle and body-wall musculature. No clear calorie distinction exists between nematodes with a parasitic or free-living habit or between large and small nematodes. PMID- 19300685 TI - Plant-parasitic Nematode Distributions in an Alfalfa Field. AB - A 7-ha alfalfa field (Medicago saliva L. cv Mesa Sirsa) was sampled systematically on a 6 x 6-m grid by removing individual cores (2.54 cm diam) to a depth of 45 cm from each of the 1,936 grid intersections. The soil was mainly coarse-textured with a fine-textured streak running centrally, north to south. Nematodes were extracted by a semiautomatic elutriator and sugar flotation sieving technique. Five plant-parasitic species were consistently present: Meloidogyne arenaria, Pratylenchus minyus, Merlinius brevidens, Helicotylenchus digonicus, and Paratrichodorus minor. All species had a highly skewed nonnormal frequency distribution that departed significantly from randontness. Goodness-of fit tests on the distribution of five populations in the entire field showed that three (Meloidogyne, Merlinius, and Helicotylenchus) were described by a negative binomial. When the samples were categorized by soil texture (coarse vs. fine textured), all populations in the fine-textured areas, and three populations (Meloidogyne, Pratylenchus, and Merlinius) in the coarse areas, fitted a negative binomial distribution. Nearly all populations titted a negative binomial when the frequency distributions from randomly located one-meter-square areas were examined for each species. PMID- 19300686 TI - Histopathogenesis of the Galls Induced by Nothanguina phyllobia in Solanum elaeagnifolium. AB - The histopathogenesis of the foliar galls induced by Nothanguina phyllobia Thorne in Solanum elaeagnilolium Cav. was examined via serial sections prepared from plant shoots at 11 time intervals (0.5-30 days) following inoculation. Nematodes infected the blades and petioles of young leaves surrounding the shoot apex. Hypertrophy and hyperplasia of the palisade, pith, cortical, and vascular parenchyma resulted in the formation of confluent leaf, petiole, and stem galls up to 25 cm(3) in volume. Externally, leaf galls were irregular, light-green, convoluted spheroid bulges distending the abaxial surface. Mature galls contained a cavity lined with parenchymogenous nutritive tissue comprising intercellular spaces and actively dividing hypertrophied cells. These cells contained granular cytoplasm, hypertrophied nuclei, and brightly stained large nucleoli. Vascular tissues were not discernibly affected during the early stages of gall development. As gall development progressed, however, vascular elements were often displaced and disoriented. The histopathology of this nematode indicates that N. phyllobia is a highly specialized parasite and, for that reason, is suitable as a biological control agent. PMID- 19300687 TI - Graphic techniques for illustrating distribution data. PMID- 19300688 TI - The development of nematology on a world basis. AB - Nematology on a world basis has experienced phenomenal growth during the last 25 years. Major influences responsible for this growth are discussed. Education of nematologists has been most evident in only a few developed nations. Some developing countries are now beginning to train nationals as nematologists. Research programs in developed countries are more intensive than in developing nations, largely because of available resources and priorities given to solving nematological problems. Developed nations have been able to take advantage of technological advances almost immediately, whereas in developing nations the lack of resources and constraints imposed by certain social or political conditions has made this difficult. Indications are that emphasis in training nematologists in developed countries may have reached a plateau, while in developing nations provisions for training and research are on the increase. PMID- 19300689 TI - Nematology-Status and Prospects: Let's Take Off Our Blinders and Broaden Our Horizons. AB - Some aspects of current fundamental nematological research and teaching in agriculture, soil zoology, biology and medicine, and parasitology are explored as they relate to the Society of Nematologists and the science of nematology in its broadest sense. PMID- 19300690 TI - Nematology-status and prospects: practical implementation of quantitative approaches to nematology. AB - The main components of the decision process in nematode pest management are the value of the predicted damage and the cost of the management alternative. The relationships involved are affected by such environmental parameters as soil texture and physiographic region. There is a general intuitive understanding of the nature of the relationships and of the effects of the environmental parameters. Some nematode damage functions have been developed through quantitative research. A conceptual framework is developed herein which promotes rational use of available experimental results, supplemented by intuitive understanding of nematodes and crops, in arriving at a pest management decision. Gaps in available data point the need for additional research within the framework. The approach allows and encourages immediate implementation. Interactive computer programs are seen as a potential vehicle for weighting the variables involved in the decision and for storing, manipulating, and delivering the necessary data funcand information. PMID- 19300691 TI - Nematology-status and prospects: the role of nematology in integrated pest management. AB - Integrated pest management (IPM) is an interdisciplinary science dealing with the development, evaluation, and implementation of pest control strategies that result in favorable economic, ecologic, and sociologic consequences. IPM has received considerable attention during the past few years, and this has led to recommendations directly related to the growth of the science of hematology. This report describes the current state of IPM in relation to the role of hematology, with special emphasis on scientific personnel requirements. All current indications are that IPM will continue to grow, very likely at an increased rate. This will place additional research, extension, and teaching demands on current hematology programs and should result in an expended resource base for nematology. PMID- 19300692 TI - Interaction of Uromyces phaseoli and Meloidogyne incognita on Bean. AB - Uromyces phaseoli, the causal agent of bean rust, suppressed shoot and root growth of three bean cultivars, reducing root weight more than shoot weight. The greatest suppression of root weight was on the cultivar that appeared most susceptible by visual ratings of shoot symptoms. Meloidogyne incognita suppressed shoot and root growth of all test cultivars; root weight reductions differed among cultivars identical in susceptibility to this pathogen in root-gall rating tests. Infection of plants with both pathogens suppressed plant weights significantly more than did infection by either pathogen alone, evidencing an additive effect. U. phaseoli and M. incognita on the same plant influenced the reproduction of one another, presumably through effects on the host. Fungal uredia were reduced in size and sporulation capacity; M. incognita produced fewer root galls, and fewer eggs per egg mass. PMID- 19300693 TI - Effects of nematicides, lime, and herbicide on peach tree short life in georgia. AB - Peach tree mortality was 75% five years after planting on a site associated with peach tree short life and receiving no nematicide treatment, no lime, and with cultivation for weed control. Mortality was reduced to 29% by preplanting plus postplanting applications of DBCP (1,2-dibromo-3-chloropropane) and with herbicidal weed control. Preplanting applications of nematicides alone did not effectively reduce tree mortality or increase yield. Lime applications increased yield but did not affect tree growth or survival. Survival was higher with weed control by a herbicide than with control by disk cultivation. Populations of Macroposthonia xenoplax were correlated positively with tree mortality and negatively with yield. The other nematode consistently present at the site, Tylenchorhynchus claytoni, was not associated with either tree mortality or yield. PMID- 19300694 TI - Species of Pratylenchus Associated with Solanum tuberosum cv Superior in Ohio. AB - Seventy-three Ohio fields comprising ca. 440 ha of cv Superior potatoes were surveyed in 1977 for plant-parasitic nematodes. Of eight genera of plant parasitic nematodes, Pratylenchus was found most frequently, occurring in 65% of the soil samples and 84% of the root samples. Populations of Pratylenchus were consistently higher than populations of the other nematode genera. The six species of Pratylenchus extracted from potato roots, in descending order of frequency, were P. crenatus, P. penetrans, P. scribneri, P. alleni, P. thornei, and P. neglectus. Prevalence of these Pratylenchus species in Ohio potato fields suggests that they could be involved with vascular wilt fungi in premature death of cv Superior potato vines known in Ohio as "early dying." PMID- 19300695 TI - Effects of Temperature on Rate of Feeding of the Plant Parasitic Nematodes Rotylenchus robustus, Xiphinema diversicaudatum, and Hemicycliophora conida. AB - Rotylenchus robustus, Xiphinema diversicaudatum, and Hemicycgiophora conida were observed feeding over a range of temperatures on perennial rye-grass (Lolium perenne) seedlings grown on agar plates. R. robustus fed between 0.5 and 42.5 C, X. diversicaudatum between 5.0 and 37.0 C and H. conida between 5.0 and 34.0 C. Between 10 and 25 C there was a direct relationship between temperature and rate of esophageal bulb contractions. Above 25 C the number of esophageal contractions/min did not increase at the same rate and eventually decreased. At the extremes of temperature range, abnormal feeding behaviour was observed. Rates of esophageal bulb contraction did not differ in the different nematode life stages and sexes, or at different feeding sites on the roots. PMID- 19300696 TI - Inhibition of Syncytia Formation and Root-knot Nematode Development on Cultures of Excised Tomato Roots. AB - Two different defined growth media were used to culture aseptically the root-knot nematode, Meloidogyne incognita, on excised roots of tomato, Lycopersicon esculentum cv 'Marglobe.' One of these media, STW, was a formulation by Skoog, Tsui, and White and the other, MS, a formulation by Murashige and Skoog. From 1 through 4 weeks, inoculated tissues were fractured to observe root infection, giant-cell formation, and nematode development with the scanning electron microscope (SEM). Four weeks after inoculation, the fresh weights of roots and developmental stages of nematodes were recorded. SEM observations indicated that roots cultured on the STW medium had normal growth and infection sites with galls that supported the development of mature females by 4 weeks. Roots cultured on the MS medium were less vigorous and had infection sites with galls containing only one to four syncytialike cells that did not support the development of mature females. Eighty percent of the larvae infecting roots cultured on the MS medium failed to develop into mature females. To determine which factor(s) affected root growth and nematode development, inoculated and uninoculated roots were grown on media consisting of different combinations of the organic and inorganic fractions of the STW and MS formulations. These experiments indicated that the organic fraction of STW was essential for normal root growth; however, the inorganic fraction of MS inhibited normal gall formation and nematode development. Further testing of the inorganic fractions revealed that the high concentration of ammonium nitrate in the MS medium was a factor that inhibited giant-cell formation and nematode development. PMID- 19300697 TI - Multiyear Population Dynamics of Ditylenchus dipsaci Associated with Phlox subulata. AB - Field population densities of Ditylenchus dipsaci associated with shoot tissue of Phlox subulata were monitored during two consecutive growing seasons and intervening periods of overwintering and plant storage. The population density increased significantly through four peaks during the first growing season, and decreased significantly during storage at 5-7 C or overwintering in the field. During the second growing season, there was only a single increase to a moderate population density, followed by a severe population decline associated with the poor physiological condition of the host. A simple model is proposed to explain the population dynamics of D. dipsaci during the first growing season. PMID- 19300698 TI - Redescription and Life History of Contortylenchus brevicomi, a Parasite of the Southern Pine Beetle Dendroctonus frontalis. AB - Larval and adult life stages are described for Contortylenchus brevicomi (Massey) Ruhm parasitizing a Mississippi population of Dendroctonus frontalis, the southern pine beetle. Fourth-stage larvae and free-living adult females of this species are identified and described for the first time. The life cycle of C. brevicomi can be reconstructed from this study. The adult female nematode lays eggs in a mature beetle. Larval development progresses within the hemocoel until fourth-stage larvae exit the host. Mating occurs in beetle galleries and only females enter an immature beetle host. PMID- 19300699 TI - Biocontrol: fungi as nematode control agents. AB - The fungal antagonists of nematodes consist of a great variety of organisms belonging to widely divergent orders and families of fungi. They include the nematode-trapping fungi, endoparasitic fungi, parasites of nematode eggs and cysts, and fungi which produce metabolites toxic to nematodes. The diversity, adaptations, and distribution of nematode-destroying fungi and taxonomic problems encountered in their study are reviewed. The importance of nemato-phagous fungi in soil biology, with special emphasis on their relationship to populations of plant-parasitic nematodes, is considered. While predacious fungi have long been investigated as possible biocontrol agents and have often exhibited spectacular results in vitro, their performance in field studies has generated little enthusiasm among nematologists. To date no species has demonstrated control of any plant pest to a degree achieved with nematicides, but recent studies have provided a much clearer concept of possibilities and problems in the applied use of fungal antagonists. The discovery of new species, which appear to control certain pests effectively under specific conditions, holds out some promise that fungi may be utilized as alternatives to chemical control after a more thorough and expanded study of their biology and ecology. PMID- 19300700 TI - Biocontrol: fungal parasites of female cyst nematodes. AB - Three species of fungi, Catenaria auxiliarls (Kuhn) Tribe, Nematophthora gynophila Kerry and Crump, and a Lagenidiaceous fungus have been found attacking female cyst nematodes. All are zoosporic fungi which parasitize females on the root surface, cause the breakdown of the nematode cuticle, and prevent cyst formation. Their identification and some aspects of their biology are reviewed. N. gynophila is widespread in Britain and reduces populations of the cereal cyst nematode, Heterodera avenae Woll., to nondamaging levels. The potential of these nematophagous fungi as biocontrol agents is discussed. PMID- 19300701 TI - Biocontrol: Bacillus penetrans and Related Parasites of Nematodes. AB - Bacillus penetrans Mankau, 1975, previously described as Duboscqia penetrans Thorne 1940, is a candidate agent for biocontrol of nematodes. This review considers the life stages of this bacterium: vegetative growth phase, colony fragmentation, sporogenesis, soil phase, spore attachment, and penetration into larvae of root-knot nematodes. The morphology of the microthallus colonies and the unusual external features of the spore are discussed. Taxonomic affinities with the actinomycetes, particularly with the genus Pasteuria, are considered. Also discussed are other soil bacterial species that are potential biocontrol agents. Products of their bacterial fermentation in soil are toxic to nematodes, making them effective biocontrol agents. PMID- 19300702 TI - Biocontrol: the potential of entomophilic nematodes in insect management. AB - A review of the development of entomophilic nematology and a commentary on the potential of entomophilic nematodes in controlling insect pests. The paper considers some of the major contributions to our knowledge of entomophilic nematology; factors involved in insect pest management and how they are applicable to the use of nematodes; nematodes which are most promising as biological control agents; and problems to be solved to facilitate the use of entomophilic nematodes in insect management. PMID- 19300703 TI - Effect of the Nematode Contortylenchus brevicomi on Gallery Construction and Fecundity of the Southern Pine Beetle. AB - Field-collected Dendroctonus frontalis were reared in a controlled environment. Male-female beetle pairs retrieved from galleries 1, 2, or 5 wk after introduction into pine bolts were examined for nematode parasites. Data were obtained for each pair on gallery length, egg niche construction, egg viability, and progeny survival. In a separate study, beetle pairs were reared under laboratory conditions for 10 wk. The number of emerged adult progeny of each pair was recorded. Contortylenchus brevicomi, a nematode parasite, was found in 25% of all beetles that established galleries. After 2 and 3 wk, female beetles infected with the nematode had produced fewer eggs and shorter galleries than did uninfected females. Uninfected females mated with nematode-infected males showed similar trends, although the differences in the 2- and 3-wk tests were not significant. Progeny survival or egg viability was not affected by nematode parasitism of either parent beetle. Unikaryon minutum, a microsporidian parasite found in 65% of all colonizing beetles, had no effect on measured variables. The lower fecundity of beetles parasitized by C. brevicomi continued throughout the insect's reproductive cycle. After 10 wk, nematode-infected beetle pairs produced fewer emerged adult progeny than did uninfected pairs. PMID- 19300704 TI - A Classification of Tylenchulus semipenetrans Biotypes. AB - The presence of two biotypes of the citrus nematode (Tylenchulus semipenetrans) in Italian citrus and olive orchards has been confirmed by comparing host specificity. Host reaction to California biotypes C1 and C3 and to three populations from Arizona, Texas, and Florida indicates that of these five United States biotypes, all except C3 consistently fit biotype C1. These findings, and the results of host-range studies in other countries, show that four biotypes of T. semipenetrans are distributed worldwide: the "Poncirus biotype," the "Citrus biotype," the "Mediterranean biotype," and the "Grass biotype." PMID- 19300705 TI - Interrelationship of Meloidogyne hapla and Ditylenchus dipsaci on Resistant and Susceptible Alfalfa. AB - Simultaneous inoculations of alfalfa with Meloidogyne hapla larvae and Ditylenchus dipsaci at 16, 20, 24, and 28 C did not depress penetration of either nematode in 'Nev Syn XX' -a selection resistant to M. hapla and D. dipsaci, 'Vernal 298' -a selection resistant to M. hapla and susceptible to D. dipsaci, 'Lahontan' -a cultivar resistant to D. dipsaci and susceptible to M. hapla, and 'Ranger' -a cultivar susceptible to both M. hapla and D, dipsaci. Infection with D. dipsaci depressed growth of susceptible 'Vernal 298' and 'Ranger' at all soil temperatures, except for 'Vernal 298' at 16 C. Infection with M. hapla alone did not depress growth of any of the alfalfas. A combination of M. hapla and D. dipsaci resulted in a synergistic weight depression on 'Ranger' at all soil temperatures. Inoculation of the four alfalfas with D. dipsaci 2, 4, 6, and 8 wk before inoculation with M. hapla at 16, 20, 24, and 28 C did not influence the resistance or susceptibility of 'Nev Syn XX,' 'Lahontan,' or 'Ranger.' However, galling of 'Vernal 298' by M. hapla was affected by soil temperature, plant age, and inoculation with D. dipsaci. PMID- 19300706 TI - Effects of one and two applications of nematicides on nematode populations and soybean yields. AB - Yields of 'McNair 800' soybeans, Glycine max (L.) Merr., were significantly increased with ethylene dibromide + chloropicrin, DBCP, phenamiphos, and aldicarb applied at-planting and with phenamiphos, aldicarb, and DBCP applied postplant to soil infested with Meloidogyne incognita (Kofoid and White) Chitwood. Yields of 'GaSoy 17' were significantly increased with ethylene dibromide + chloropicrin, DBCP, phenamiphos, and aldicarb applied, preplant and with DBCP, carbofuran, phenamiphos, aldicarb, and DBCP applied postplant to soil infested with Hoplolaimus columbus Sher. In several instances, preplant or at-planting treatments plus postplant treatments with the same or different chemicals were more effective than either treatment alone. Generally, the fumigants were more effective than the nonfumigants when they were applied at-planting to M. incognita-infested soil and preplant to H. columbus-infested soil. Phenamiphos, aldicarb, and DBCP were about equally effective when they were applied postplant in M. incognita-infested soil, but DBCP was more effective than carbofuran. Carbofuran, phenamiphos, aldicarb, and DBCP were about equally effective when applied postplant to H. columbus-infested soil. PMID- 19300707 TI - Morphological comparison of meloidogyne female head structures, perineal patterns, and stylets. AB - The external morphology of female heads of three populations of each of two cytological races of Meloidogyne hapla (race A-meiotic, race B-mitotic) and single populations of M. arenaria, M. incognita, and M. javanica was compared by light (LM) and scanning electron microscopy (SEM). Perineal patterns of all nine populations were observed with a LM and then examined with a SEM. In addition, female stylets of each population were excised, viewed with a SEM, and compared with observations made with a LM. Head morphology of the females, including shape of medial and lateral lips, expression of sensilla, and head annulation, was distinct for each species, each race of M. hapla, and each population of M. hapla race A. The morphology of a given perineal pattern appeared similar with the SEM and the LM. The SEM emphasized surface details, whereas the LM revealed subcuticular structure as well. Stylet morphology was unique for each species but similar in all populations of M. hapla. There were differences between species in the shape of the cone, shaft, and knobs and in the distance of the dorsal esophageal gland orifice from the stylet knob base. Several of the morphological characters first detected in the SEM were seen subsequently with the LM and are helpful in species identification. PMID- 19300708 TI - Population Increase of Pratylenchus hexincisus on Corn as Related to Soil Temperature and Type. AB - Population increase of Pratylenchus hexincisus on corn was tested over 3 months at 15, 20, 25, and 30 C in Marshall silt loam, Clarion silt loam, Buckner coarse sand, and Haig silty clay loam soils. The optimum temperature for increase was 30 C in all soils. The nematode population was significantly larger in Buckner coarse sand than in other soil types at 50 C. The recovered P. hexincisus populations equaled or exceeded initial inoculum levels at the two higher temperatures in Marshall silt loam and Haig silty clay loam and at 30 C in Clarion silt loam and Buckner coarse sand. P. hexincisus required 32,400 heat units in Haig silty clay loam and more than 40,000 heat units in the three other soil types to reach a level that is known to cause significant height and biomass reduction in corn under controlled condition. PMID- 19300709 TI - Description and SEM Observations of Meloidogyne chitwoodi n. sp. (Meloidogynidae), a Root-knot Nematode on Potato in the Pacific Northwest. AB - Meloidogyne chitwoodi n. sp. is described and illustrated from potato (Solanum tuberosum) originally collected from Quincy, Washington, USA. This new species resembles M. hapla, but its perineal pattern is basically round to oval with distinctive and broken, curled, or twisted striae around and above the anal area. The vulva is in a sunken area devoid of striae. Vesicles or vesicle-like structures are present in the median bulb of females. The larva tail, being short and blunt with a hyaline tail terminal having little or no taper to its rounded terminus, is distinctively different from M. hapla. SEM observations revealed the nature of the perineal pattern and details of the head of larvae and males, and showed the spicules to have dentate tips ventrally. Hosts for M. chitwoodi n. sp. include potato, tomato, corn, and wheat but not strawberry, pepper, or peanut. The latter three crops are excellent hosts for M. hapla. The known distribntion of this new root-knot species presently involves certain areas of Idaho, Washington, and Oregon. The common name "Columbia root-knot nematode" is proposed for M. chitwoodi n. sp. PMID- 19300710 TI - Measurement of the Vertical Migration of Anguina tritici (Steinbuch 1799) Chitwood, 1935 in Soil under Experimental Conditions. PMID- 19300711 TI - Isolation of Strelkovimermis peterseni, a Mermithid Parasite of Anopheline Mosquitoes in Northeastern New York. PMID- 19300712 TI - Instar Susceptibility of Simulium vittatum (Diptera: Simuliidae) to the Entomogenous Nematode Neoaplectana carpocapsae. AB - Laboratory bioassays showed that the susceptibility of Simulium vittatum to Neoaplectana carpocapsae increased with successive larval instars. First, second, and third instar larvae were resistant to infection, while seventh instars were highly susceptible. Significant differences in intra-instar susceptibility were also evident, as mortality ranged from 58% for the smallest seventh instar larvae to 97% for the largest. Dissections revealed that the basis for the resistance of early instars was physical exclusion of the comparatively large nematodes. The principle factor regulating the susceptibility of mid and late instars was injury to nematodes caused by larval mouthparts during ingestion. Differences in intra instar susceptibility were similarly related to nematode injury. PMID- 19300713 TI - Symptomatology and Histopathology of Soybean Roots Infected by Pratylenchus scribneri and P. alleni. AB - Size of lesions caused by Pratylenchus scribneri on roots of 'Clark 63' soybean was correlated with nematode colony size within roots. A single nematode was capable of causing a detectable lesion. When a root became highly necrotic and shrunken, few nematodes but numerous eggs remained in the tissue. In histological sections made 5, 11, 18, and 45 d after planting, P. scribneri was located entirely within the cortex and generally was oriented longitudinally to the vascular cylinder, either outstretched in the same plane or coiled through several cells. Nematodes moved intracellularly, causing extensive rupturing of cell walls, retraction and disappearance of cytoplasm, and thickening of cell walls and necrosis of cells around feeding sites. Depth of penetration within the cortex and necrosis of cells increased with time after infection, eventually resulting in formation of cavities in the cortex and occasional secondary injury to the endodermis. Stele tissue was unaffected by feeding, and damage to the epidermis was limited to nematode entry points. Orientation of P. alleni and histopathology of its infection at 45 days were identical to those of P. scribneri, except that there was no injury to the endodermis. PMID- 19300714 TI - Parthenogenesis in the Two Races of Radopholus similis from Florida. AB - Single larval inoculations of both the citrus and banana races of Radopholus similis from Florida indicate parthenogenetic reproduction is possible. These races normally reproduce amphimictieally. Both races produced all-female populations from single larva inoculations after 80 d and male-female populations after 180 d. The all-female populations produced female progeny with viable eggs, but no spermatozoa were present in the spermatheca, indicating tychoparthenogenesis occurred. Uninseminated females in the latter study produced both male and female progeny. Approximately 95% of the female progeny of the male female populations were inseminated, indicating that cross-fertilization had occurred. The proportion of males to females was as expected in a normal bisexual population. No intersex males were observed. The mechanism of sex determination is not fully understood but assumed to be enviromnentally induced. PMID- 19300715 TI - Karyology and Oogenesis of Radopholus similis (Cobb) Thorne. AB - Two morphologically indistinguishable races of Radopholus similis from Florida are presently identifiable only by host preference tests. One race attacks banana and other hosts, but not citrus. The other race attacks both citrus and banana, as well as other hosts. The two races normally reproduce by amphimixis. The morphology of the female reproductive systems of both races were examined and appeared to be identical. However, slight differences were observed in developmental stages of oogenesis between the two races. The two races were distinct with respect to karyotype. There were four chromosomes in the banana race and five chromosomes i n the citrus race. Karyotypic uniformity was found i n the two Florida populations of the citrus race. PMID- 19300716 TI - Effect of Foliar Application of Carbofuran and a Related Compound on Plant parasitic Nematodes under Greenhouse and Growth Chamber Conditions. AB - Greenhouse and growth chamber studies were made to investigate the downward systemic nematicidal activity of carbofuran and its analog 2,3-dihydro-2,2 dimethyl-7-benzofuranyl [(di-n-butyl)-4-aminosulfenyl] carbamate against Meloidogyne incognita, Tylenchorhynchus claytoni, and Pratylenchus penetrans. Oxamyl was used as standard in tests with T. claytoni. Both carbofuran and its analog reduced all three plant-parasitic species when plant foliage was sprayed with chemical suspension of 1,200, 2,400 or 4,800 ppm. The studies show that fewer chemical applications were required to reduce populations of P. penetrans than to reduce populations of T. claytoni. Oxamyl was somewhat less active than either carbofuran or its analog. PMID- 19300717 TI - Action of Systemic Nematicides in Control of Xiphinema iindex on Grape. AB - In greenhouse tests using potted grape plants three nematicides, aldicarb 10 G at 4.5 ai/ha, phenamiphos 15 G at 22 kg ai/ha, and oxamyl liquid at 4.5 kg ai/ha, were tested against Xiphinema index on 'Thompson Seedless' grape. Different timings for chemical treatments and X. index inoculations were used to determine some of the aspects of the mode of action. When nematodes and nematicides were applied simultaneously, nematodes were reduced from the initial 500 to the averages 5, 1, and 4, respectively, for aldicarb, phenamiphos, and oxamyl. Similar counts (respectively, 3, 1, and 2) were obtained when the nematicides were added first and the nematodes 14 d later. Nematode counts were 83, 112, and 1,346 when nematicides were applied first, and 14 days later plants were washed free of soil, repotted in untreated soil, and then inoculated. In untreated controls the population increased to an average of 2,703. Plant growth was inversely related to the level of nematode population resulting from the treatment. PMID- 19300718 TI - Residue dynamics and persistence of aldicarb and its biologically similar active metabolites in grapevines. AB - Residue dynamics in giapevine of the nematicide aldicarb (2-methyl-(metbylthio) propionaldehyde-O-(Methylcarbamoyl) oxime) and its biologically similar active metabolites, aldicarb sulfoxide and aldicarb sulfone, were determined by gas chromatographic techniques. Residues were found in the roots, trunks, stems, and leaves of grapevine 120 d after application. Residues in leaves as high as 1.40 and 8.89 ppm resulted from 4.5 and 9 kg ai/ha respectively. In roots, trunks, and stems the residues had also declined after 180 d. No residues were detected in the newly forming immature fruit. Residues in roots, trunks, young branches, and leaves declined further after 270 d, but residues in mature fruit at harvest time were 0.03 and 0.05 ppm from application of 4.5 and 9 kg ai/ha, respectively. In other trials the amount of aldicarb toxic residues found in mature fruit at harvest time varied with grape varieties, time and rate of application, total amount of rainfall, irrigation water, and soil type. PMID- 19300719 TI - Interaction of fensulfothion and phorate with preemergence herbicides on soybean parasitic nematodes. AB - The herbicides alachlor, linuron, vernolate, and metribuzin were applied to plots treated with the nematicide fensulfothion or the insecticide phorate and planted to soybean in two locations in North Carolina. In 1976 treatment with fensulfothion + alachlor or vernolate, phorate + alachlor or metribuzin resulted in greater nematode population densities than no treatment, or treatment with fensulfothion alone, or phorate alone. In 1977 fensulfothion and phorate alone and in combination with the preemergence herbicides effectively controlled Tylenchorhynchus cIaytoni. Late season population resurgence of Heterodera glycines occurred in fensulfothion + alachlor treated plots. Correlation coefficients for H. glycines vs. yield were -0.48 (P = 0.05) and -0.46 (P = 0.05) for 30 and 68 d after planting, respectively. PMID- 19300720 TI - Dynamics of Concomitant Populations of Hoplolaimus columbus, Scutellonema brachyurum, and Meloidogyne incognita on Cotton. AB - Cotton seedlings grown in a greenhouse and a growth chamber were inoculated with Scutellonema brachyurum, Hoplolaimus columbus, and Meloidogyne incognita, singly and in all possible combinations, at two initial population (Pi) levels (100 and 300/100 cm(3)). S. brachyurum alone was not pathogenic to cotton at these population levels. It fed primarily as an ectoparasite but matured and reproduced within the root when it penetrated. Populations of S. brachyurum increased in the presence of H. columbus but were suppressed by M. incognita. H. columbus suppressed dry shoot weights of cotton (P = 0.05) at a Pi of 300/100 cm(3) soil. Simultaneous inoculation of H. columbus with either M. incognita or S. brachyurum increased H. columbus populations over treatments with H. columbus alone, both at 60 and 90 d after inoculation. M. incognita suppressed cotton shoot weights significantly (P = 0.05) at both Pi levels. Inoculation with S. brachyurum increased M. incognita populations 60 d after inoculation, while H. columbus suppressed populations of M. incognita. Most larvae of M. incognita did not develop to maturity in the presence of H. columbus. Giant cells aborted and were necrotic 20-25 d after inoculation. Since M. incognita and H. columbus feed on different tissues, the inhibition of M. incognita may have resulted from a physiological effect of H. columbus on the host. PMID- 19300721 TI - Influence of Aphelenchus avenae on Vesicular-arbuscular Endomycorrhizal Growth Response in Cotton. AB - The influence of Aphelenchus avenae on the relationship between cotton (Gossypium hirsutum 'Stoneville 213') and Gigaspora margarita or Glomus etunicatus was assessed by its effect on the mycorrhizal stimulation of plant growth and microorganism reproduction. The mycophagous nematode usually did not suppress stimulation of shoot growth resulting from mycorrhizae (G. margarita) at inoculum levels of 3,000 or 6,000 nematodes per pot, but retarded root growth at 6,000 per pot. When the nematode inoculum was increased to 10, 40, or 80 thousand, G. margarita stimulation of shoot or root growth was retarded at the two higher rates. Shoot growth enhancement by G. etunicatus was suppressed by 10 thousand A. avenae but not by 40 or 80 thousand. A. avenae reproduced better when the nematode was added 3 wk after G. margarita than with simultaneous inoculations. Sporulation of both fungi was affected little by the mycophagous nematode. The high numbers of A. avenae required for an antagonistic effect probably precludes the occurrence of any significant interaction between these two organisms under field conditions. PMID- 19300722 TI - Interaction Between Meloidogyne arenaria and Glomus fascicuqlatus in Grape. AB - Root zones of grape (Fitis vinifera cv Thompson Seedless) cuttings were infested with chlamydospores of Glomus fasciculatus or eggs of Meloidogyne arenaria or both. Growth of grapevines was greatest in mycorrhizal (G. fasciculatus) plants. Mycorrhizal development and growth of mycorrhizal and nonmycorrhizal plants were reduced in the presence of M. arenaria. At low initial nematode inoculum (PI) levels (approx. 200 eggs/plant), the presence of mycorrhizae enhanced plant growth during 1 yr, but no significant benefit was achieved by mycorrhizae where PI was high (approx. 2,000 eggs/plant). Final nematode populations were highest in mycorrhizal plants. PMID- 19300723 TI - Influence of Meloidogyne incognita on Fusarium Wilt of Tomato at or below the Minimum Temperature for Wilt Development. AB - 'Bonny Best' tomato plants were grown at 16, 21, or 24 C for 28 d in soil infested with either of two isolates of Fusarium oxysporum f. sp. lycopersici race 1 and Meloidogyne incognita. Significant levels of fusarium wilt occurred at all temperatures including 16 C, which has not been reported previously. One Fusarium isolate resulted in the highest levels of disease incidence at 21 and 24 C in the presence of root-knot nematodes, and at 24 C when the nematodes were not present. At 16 C there was no significant difference in the number of plants infected by the second Fusarium isolate alone or in combination with root knot nematodes, although the presence of nematodes resulted in a significant increase in the percentage of disease occurrence and vessel infection at 21 C. PMID- 19300724 TI - Identification of the Chromosome Carrying the Factor for Resistance to Meloidogyne incognita in Tobacco. AB - To identify the chromosome carrying the factor for resistance to Meloidogyne incognita in tobacco, crosses were made between resistant tobacco 'NC95' as pollen parent and each of the 12 tobacco monosomics (A-L) representative of the Tomentosae half of the Nicotiana tabacum chromosome complement. Of the F seedlings, 927 plants were grown for observation. From these, 223 plants were selected as possible monosomics on the basis of morphological characteristics. These plants were self-pollinated, and the resulting F plants were inoculated with both M. incognita acrita and M. incognita incognita. Sixteen F populations, derived from the haplo-G monosome, were completely resistant. All of the F populations derived from the other 11 monosomic crosses segregated into a 3:1 (resistant:susceptible) ratio. These results indicate that the factor for resistance to M. incognita is located on the G chromosome of N. tabacum. This is the first report establishing the N. tabacum chromosome that carries the factor for root-knot resistance. The results are consistant with our earlier evidence that M. incognita resistance in tobacco is derived from N. tomentosa, a species in the section Tomentosae of the subgenus Tabacum, genus Nicotiana. The other 12 chromosomes of N. tabacum have affinities with N. sylvestris, section Alatae, subgenus Petunoides, genus Nicotiana. PMID- 19300725 TI - Interrelationships of Meloidogyne Species with Flue-cured Tobacco. AB - Microplot and field experiments were conducted to determine relationships of population densities of Meloidogyne spp. to performance of flue-cured tobacco. A 3-yr microplot study of these interactions involved varying initial nematode numbers (P(i)).and use of ethoprop to re-establish ranges of nematode densities. Field experiments included various nematicides at different locations. Regression analyses of microplot data from a loamy sand showed that cured-leaf yield losses on 'Coker 319' for each 10-fold increase in P(i) were as follows: M. javanica and M. arenaria--13-19%; M. incognita-5-10%; M. hapla-3.4-5%; and 3% for M. incognita on resistant 'Speight G-28' tobacco. A P(i) of 750 eggs and larvae/500 cm(3) of soil of all species except M. hapla caused a significant yield loss; only large numbers of M. hapla effected a loss. M. arenaria was the most tolerant species to ethoprop. Root-gall indices for microplot and most field-nematicide tests also were correlated negatively with yield. Relationships of P(i)(s) and necrosis indices to yield were best characterized by linear regression models, whereas midseason numbers of eggs plus larvae (P(m)) and sometimes gall indices vs. yield were better characterized by quadratic models. The relation of field P(m) and yield was also adequately described by the Seinhorst model. Degrees of root galling, root necrosis, yield losses, and basic rates of reproduction on tobacco generally increased from M. hapla to M. incognita to M. arenaria to M. javanica. PMID- 19300726 TI - Sclerostylus n. gen. from Panama and Other Neotropical Species of LeptonIchoidea (Dorylaimida). AB - Leptonchoid species found in soil samples taken in lowland and mid-elevation seasonal forests at four locations in Panama were as follows: Sclerostylus karri n. gen., n. sp., Basirotyleptus saccatus n. sp., B. nemoralis, B. acus, B. penetrans, Doryllium neotropicum n. sp., D. minor, Tyleptus gymnochilus, Gerthus jamesi, Dorylaimoides micoletzkyi, and Tylencholaimellus macrophallus. The discoveries corroborate our earlier suggestion of a Gondwanian origin for Tyleptus and Basirotyleptus and provide data for additional biogeographical study. PMID- 19300727 TI - Parasitic Habits of Ogma rhombosquamatum and Description of the Male. PMID- 19300728 TI - Invasion of Sugarbeet Leaf Blades by Larvae of Heterodera schachtii. PMID- 19300729 TI - Erratum. AB - [This corrects the article on p. 328 in vol. 12.]. PMID- 19300730 TI - Oogenesis and the Chromosomes of the Parthenogenic Root-knot Nematode Meloidogyne incognita. AB - 220 populations of Meloidogyne incognita and related forms from 46 countries reproduced by mitotic parthenogenesis (apomixis). Determination of somatic chromosome numbers from oogonia and oocytes revealed the existence of a predominant, possibly triploid race A with 3n = 40 to 46 and a rare, diploid race B with 2n = 32 to 36 chromosomes. There is no correlation between cytological races and the four recognized host races of this species. The characteristic behavior of prophase I chromosomes of maturing oocytes, which results in a prolonged prophase stage, is a unifying feature of all forms of M. incognita and supports monophyletic evolution, distinct from that of other Meloidogyne species. Extensive chromosomal polymorphism detected among populations can be helpful in elucidating the cytological pathway of evolution of the species. PMID- 19300731 TI - Diacyl, Alkylacyl, and Alkenylacyl Phospholipids of Meloidogyne javanica Females. AB - The phospholipid composition and acyl, alkyl, and alkenyl group compositions of diacyl, alkylacyl, and alkenylacyl phosphoglycerides of M. javanica were investigated. Phospholipid was comprised of 61.7% choline phosphoglyceride, 22.0% ethanolamine phosphoglyceride, and smaller quantities of six other lipids. Phospholipid fatty acid was more unsaturated than neutral lipid fatty acid and contained 61.3% octadecenoic (18:1) acid. Fatty acid at the 1-position of diacyl phospholipids was shorter and more saturated than that at the 2-position. Compared to choline phosphoglyceride, ethanolantine phosphoglyceride contained less 18:1 and 20:5 and more 18:0 and 20:0 acid. Alkenylacyl and alkylacyl compounds comprised 34.6% and 9.3%, respectively, of the ethanolamine phosphoglyceride but only 0.5% and 0.6% of the choline phosphoglyceride. Alkenylacyl and alkylacyl ethanolamine phosphoglycerides contained a smaller percentage of 20-carbon polyunsaturated acid at their 2-positions than did their diacyl analogue. At least 95% of the alkenyl and alkyl groups were 18:0 compounds. Tomato roots did not contain alkenylacyl or alkylacyl phosphoglycerides; their occurrence in M. javanica is a significant biochemical difference between the nematode and its host. PMID- 19300732 TI - Pararotylenchus n. gen. (Pararotylenchinae n. subfam., Hoplolaimidae) with Six New Species and Two New Combinations. AB - A new subfamily, Pararotylenchinae, (Hoplolaimidae) is described. It includes a single genus, Pararotylenchus n. gen., six new species, attd two new combinations, Pararotylenchus (syn. Tylenchorhynchus) brevicaudatus (Hopper, 1959) n. comb. and Pararotylenchus (syn. Rotylenchus) pint (Mamiya, 1968) n. comb. Pararotylenchinae is similar to certain other Hoplolaimidae, such as Rotylenchinae, with respect to most characters including the short tail, the position of the phasmids near the anus, and the relatively great distance of the dorsal gland orifice from the base of the stylet knobs. The lip region, as observed with the scanning electron microscope, conforms to the basic pattern for Hoplolaimidae. The labial disc is round with slit-like amphidial openings at the lateral sides of the periphery of the disc. The distinctive anterior-most lip annule is segmented into six sectors. Unlike other Hoplolaimidae, however, the esophageal glands of Pararotylenchinae form a basal bulb similar to that of Tylenchorhynchidae; other characters do not resemble Tylencborhynchidae. Specimens of Pararotylenchus have been recovered only from cool regions at high elevations and Pacific coastal areas in the Western United States, Japan, and Korea. PMID- 19300733 TI - Description and SEM Observations of Dolichodorus marylandicus n. sp. With a Key to Species of Dolichodorus. AB - Dolichodorus marylandicus n. sp. is described and illustrated from grass (Zoysia japonica) in College Park, Maryland. Specimens have also been collected from perennial bluegrass (Poa pratensis) pasture at Beltsville, Maryland, and from pine (Pinus sp.) in North Carolina. This new species is related to D. heterocephalus Cobb, D. similis Golden, and D. aestuarius Chow &Taylor. Females are distinct in having a longer styler and shorter body length than D. aestuarius. The excretory pore is at the level of the isthmus or slightly posterior and the tail is abruptly reduced in diameter, tapering to an acuminate terminus. The tails of D. similis and D. heterocephalus conically taper to a median point, with D. similis having an especially long tail. D. marylandicus does not possess the rounded, sclerotized accessory structures adjacent to the vulva observed in lateral views of D. similis and D. heterocephalus. SEM observations of D. heterocephalus and D. marylandicus revealed details of the head of males and females, and species difference in shape of the oral disc. PMID- 19300734 TI - Description of Trilineellus clathrocutis n.g., n.sp. (Tylenchorhynchinae: Tylenchida Thorne, 1949) with a Key to Species and Observations on Tylenchorhynchus sensu stricto. AB - TrilineeIlus clathrocutis n.g., n.sp. is described and illustrated. It was found as an associate of corn (Zea mays) in Stockton, Georgia, USA, and is related to a group of Tylenchorhynchus sensu lato species having three lines in nonareolated lateral fields. This new species is closely related to Tylenehorhynehus divittatus Siddiqi 1961, T. sculptus Seinhorst 1963, and T. triglyphus Seinhorst 1963 (syn. T. chonai Sethi & Swarup 1968) Tarjan 1973. It differs from these species primarily by having longitudinal striae on the body. These four species are differentiated from Tylenchorhynchus sensu stricto by having three lateral lines instead of four. They differ from Uliginotylenchus Siddiqi 1971 by having nonareolated lateral fields, fewer than 25 annules on conoid rounded tails, differently shaped gubernacula, nonattenuated stylets, and other distinctive characters. They differ from Triversus Sher 1973 by having the male tail enclosed by the bursa and by having rounded female tails. SEM observations of T. clathrocutis reveal a cuticle deeply cut by longitudinal and horizontal striae and bearing wide (> 2.0 mum) annules. Trilineellus is proposed to accommodate the new species and the three-incisured species still within Tylenchorhynchus. Tylenchorhynchus is thereby the repository for species within Tylenchorhynchinae having four lines in the lateral field, no conspicuous labial disc, and bursa enclosing the male tail. PMID- 19300735 TI - Effects of film mulch and soil pesticides on nematodes, weeds, and yields of vegetable crops. AB - Field plots in Tifton loamy sand were treated with various soil pesticides in 1973 and 1974 and either left exposed or covered with biodegradable flint mulch. Test crops were cantaloup, slicing and pickling cucumber, squash, and sweet corn. Overhead sprinkler irrigation was used in 1973, and trickle irrigation under the film mulch was used on sweet corn in 1974. Soil was assayed for nematodes, and roots of plants were evaluated for damage by root-knot nematodes. Nematode populations were reduced by soil treatment with an organic phosphate or carbamate nematicide-herbicide-fungicide combination (NHF), DD-MENCS, methyl bromide chloropicrin (MBR-CP), ethoprop, carbofuran, and sodium azide + ethoprop or carbofuran. Sodium azide, sodium azide + ethoprop or carbofuran, ethoprop, and carbofuran were less effective than DD-MENCS, MBR-CP, attd the NHF combination. The NHF combination controlled grasses and broadteaf weeds as effectively as the herbicide alone. Growth and yield were greatest when nematodes and weeds were controlled. Yields of marketable vegetables were highest from plants in plots treated with DD-MENCS with a film mulch. PMID- 19300736 TI - Influence of water and soil temperature on the concentration and efficacy of phenamiphos for control of root-knot nematodes. AB - Field plots of Tifton loamy sand were treated with phenamiphos for control of root-knot nematodes in a multiple-crop system of turnips, field corn, and southern peas. Preplant applications of phenamiphos protected roots of turnips and corn from damage by root-knot nematodes. Concentrations of phenamiphos at application in the 0-15-cm soil layer were near 6 mug/g on turnips and near 4 mug/g on corn and southern peas. After 30 d, concentrations were approximately 1 mug phenamiphos/g of soil for all crops. Concentrations of 2.0-3.8 mug phenamiphos/g of soil for 9-d duration appeared to be adequate for control of root-knot nematodes on field corn and southern peas in this multiple-crop system. Stepwise regression analyses indicated that 31%, 62%, and 22% of the variations in concentration of phenamiphos in the soil planted to turnips, corn, and southern peas, respectively, were attributable to the amount of water that the plots received. Soil temperature had no effect on concentrations of phenamiphos. PMID- 19300737 TI - Applying Nematicides through an Overhead Sprinkler Irrigation System for Control of Nematodes. AB - Phenamiphos, ethoprop, and carbofuran each at 6.7 kg a.i./ha were applied to squash, southern pea, and corn via injection into a sprinkler irrigation system. This method was then compared with a conventional application of phenamiphos and ethoprop granules spread on the soil surface and incorporated into the top 15 cm for control of Macroposthonia ornata and Meloidogyne incognita. Nematode populations in the soil and root-gall indices were lower, and yields greater, in treated than in untreated plots, but there were no significant differences between the methods of application in most comparisons. PMID- 19300738 TI - Species of Phytophthora and Pythium as Nematode-destroying Fungi. AB - Pythium monospermum,, P. aphanidermatum, and Phytophthora palmivora were found to be capable of destroying certain nonstylet-bearing nematodes through endozoic parasitism by hyphae from ingested zoospores. Hyphae of P. monospermum parasitized nematode eggs but could not capture or otherwise prey upon living nematodes. We suggest that endoparasitism of free-living nematodes may be common among Oomycetes in nature. PMID- 19300739 TI - Influence of Nonhosts, Crucifers, and Fungal Parasites on Field Populations of Heterodera schachtii. AB - Heterodera schaehtii egg number decline under nonhosts was surveyed for 3-4 years at soil depths of 0-30 cm and 30-60 cm in three fields in the Imperial Valley, California. In the two fields continously cropped to alfalfa, annual decline rates were 49 and 63%, respectively, and did not differ (P = 0.05) between depths. In the third field, cropped to annual nonhosts and fallowed, decline rates of 56 and 80% at 0-30-cm and 30-60-cm depths, respectively, were significantly different (P = 0.05). Egg hatch is the major cause of decline. Soil moisture in relation to type of cropping sequence apparently influenced egg hatch and activity of fungal parasites. An interaction matrix is used to assess the importance of biological, environmental, and management factors affecting decline of H. schachtii egg numbers. The required rotation length to non-hosts for various egg densities can be predicted. In coastal California, inclusion of a winter crucifer crop in the rotation increased H. schachtii egg density up to threefold. PMID- 19300740 TI - Infra-species Variation in Reactions to Hosts in Heterodera glycines Population. AB - Eighteen hosts were inoculated with each of four races of Heterodera glycines. A discriminant function analysis of the reactions of these races to these hosts demonstrated that these races could be separated but not consistently. Then 33 H. glycines populations collected from 13 states and five obtained from Japan were tested on differential hosts. The number of variants discriminated within these 38 populations depended on the number of differentials and the rating system used. When five differentials were used with a (+) or (-) rating system there were six "races," but when 13 differentials were used with a (+) or (-) system there were 25 physiological groups. If an index rating system was used there were 36 groups. Apparently H. glycines is a very variable species and delineation of races varies with criteria chosen. PMID- 19300741 TI - The Relationship of Heterodera schachtii Population Densities to Sugarbeet Yields. AB - Sugarbeet yields were contpared with field populations of Heterodera schachtii Schmidt. The correlation between sugarbeet yields and viable larvae/g of soil was negative and high, but that between sugarbeet yields and viable cysts/g of soil was lower. Sugarbeet yields were also compared with H. schachtii populations by years of rotation with a nonhost crop. The coefficients of correlation (r) between yield and viable larvae/g of soil were negative and high: 0 yr of rotation, -0.935; 1 yr, -0.922; 2 yr, -0.954; 3 yr, -0.935; and combined years, 0.965, with 95% confidence limits of -0.91 to -0.98 for combined years. The comparable correlation coefficients between yield and "viable cysts"/g of soil were negative and lower: 0 yr of rotation, -0.151; 1 yr, -0.022; 2 yr, -0.490; 3 yr, -0.456; and combined years, -0.586, with 95% confidence limits of -0.22 to 0.80 for the combined years. PMID- 19300742 TI - The Relationship of Plant Age, Soil Temperature, and Population Density of Heterodera schachtii on the Growth of Sugarbeet. AB - There were direct relationships between inoculum density of Heterodera schachtii Schm. (nematode population density), initial soil temperature, the growth of sugarbeets in the greenhouse under controlled temperatures, and nematode populations. Heterodera schachtii was least pathogenic on plants inoculated at 6 wk of age and most pathogenic on plants grown from inoculated germinated seed (0 wk of age). In the field, H. schachtii was least pathogenic on sugarbeets grown at an initial soil temperature of 6 C and most pathogenic on those grown at an initial soil temperature of 24 C. The growth period for sugarbeets at the different soil temperatures was determined by heat units; since penetration of sugarbeet roots by H. schachtii larvae is accelerated at soil temperatures above 10 C, each hour-degree ahove 10 C was counted as one effective heat unit (HU). Using this guideline it was determined that root weight depressions in the greenhouse, for each degree-unit population (HU-UP) where unit population = one larvae/g soil, were 0.052, 0.09, 0.12, and 0.17 mg at initial soil temperatures of 6, 12, 18, and 24 C, respectively. Root weight depressions were 0.28, 0.23, 0.15, and 0.086 mg when plants were inoculated at 0, 2, 4, and 6 wk of age. PMID- 19300743 TI - Pathological Differences in Heterodera schachtii Populations. AB - Five populations of Heterodera schachtii Schm. from Oregon, Idaho, and Utah did not differ significantly in seedling penetration and rate of emergence and virulence. Another Utah H. schachtii population (Utah 2), however, differed from these five populations in all of the above-mentioned characteristics. More H. schachtii larvae of the Utah 2 population than the other populations penetrated sugarbeet seedlings at 10, 15, 20, and 25 C. Root and top weights of sugarbeet plants were signiticantly less when roots were parasitized by the Utah 2 population than when they were parasitized by larvae of the other nematode populations under similar experimental conditions. Also, the period of larval emergence was shorter in the Utah 2 population than in any of the other H. schachtii populations. PMID- 19300744 TI - Yield Response and Injury Levels of Meloidogyne incognita and M. javanica on the Susceptible Tobacco 'McNair 944'. AB - The effects of Meloidogyne incognita and M. javanica on a susceptible tobacco (Nicotiana tabacum L.) cv. McNair 944 were investigated in field microplots during 1978 and 1979. Three initial inoculum levels-4, 16, and 64 nematode eggs and/or second-stage larvae per 100 cm(3) of soil-were used for each nematode species. Data obtained from the experiments included plant yield and the amount of reproduction of the two nematode species. At comparative inoculum levels, M. javanica was more aggressive than M. incognita on tobacco and caused approximately twofold more yield suppression than M. incognita. The calculated initial population of M. incognita, derived from the average for 2 yr, which produced a 7% suppression in plant yield was four eggs and/or second-stage larvae per 100 cm(3) of soil; whereas less than one M. javanica egg and/or second-stage larvae per 100 cm(3) of soil was needed to achieve similar suppression. Nematode reproduction varied in the 1978 and 1979 tests, but similar trends were observed. Early season M. javanica populations were greater than those of M. incognita, but late season populations of M. incognita were twice anti three times those of M. javanica. PMID- 19300745 TI - Comparative Studies on Root Invasion, Root Galling, and Fecundity of Three Meloidogyne spp. on a Susceptible Tobacco Cultivar. AB - Root invasion, root galling, and fecundity of Meloidogyne javanica, M. arenaria, and M. incognita on tobacco was compared in greenhouse and controlled environment experiments. Significantly more M. javanica than M. arenaria or M. incognita larvae were found in tobacco roots at 2, 4, and 6 d after inoculation. Eight days after inoculation there were significantly more M. arenaria and M. javanica than M. incognita larvae. Ten days after inoculation no significant differences were found among the three Meloidogyne species inside the roots. Galls induced by a single larva or several larvae of M. javanica were significantly larger than galls induced by M. incognita: M. arenaria galls were intermediate in size. Only slight differences in numbers of egg masses or numbers of eggs produced by the three Meloidogyne species were observed up to 35 d after inoculation. PMID- 19300746 TI - Host-Parasite Interaction of Resistant Sugarbeet and Heterodera schachtii. AB - The host-parasite relationships between Heterodera schachtii Schm. and the nematode-resistant diploid Beta vulgaris L. line '51501' were examined via serial sections of secondary rootlets. Second-stage larvae penetrated sugarbeet roots and migrated up to 1.95 mm before establishing permanent feeding sites. Most sedentary larvae were oriented parallel to the root axis or in various diagonal or folded positions in the cortex. Nematodes adopted no definite orientation with regard to the root apex. Nematode feeding stimulated formation of multinucleate syncytia in host tissues. Syncytia were 0.3-1.1 mm in length, up to 90 [mu]m x 150 [mu]m in cross section. Root diameters were enlarged close to feeding sites. Usually nematodes deteriorated concomitant with necrosis of syncytia, and dead nematodes frequently appeared macerated or flattened and deformed. Most nematodes did not develop to maturity" in the resistant host tissues, Cavities left by collapse of syncytia were filled by growth of parenchymatous tissue. PMID- 19300747 TI - Effect of Soil Texture and the Clay Component on Migration of Meloidogyne incognita Second-stage Juveniles. AB - The vertical migration of M. incognita juveniles introduced at 20 cm from the roots was studied in five natural soils, 100% silica sand, 95% silica sand with 5% clay, 90% silica sand with 10% clay, and 95% silica sand with 5% clay as a concentrated layer. In natural soils the percentage of juveniles capable of migrating 20 cm and penetrating the roots decreased when the percentage of clay and silt increased. No migration occurred in silica sand without clay particles; when 5 or 10% of clay were mixed to silica sand, 34 and 26%, respectively, of the juveniles were able to migrate 20 cm. Clay separated from silica sand in which tomatoes were grown was attractive for juveniles. It is suggested that clay particles aid in the migration of root-knot juveniles over long distances to plant roots by absorbing and holding root exudates or bacterial by-products which form a concentration gradient enabling nematodes to locate roots. PMID- 19300748 TI - Influence of photoperiod and temperature on migrations of meloidogyne juveniles. AB - Photoperiod influences the migration of M. incognita juveniles toward tomato roots. Approximately 33% migrated vertically 20 cm in 7 days to roots when 12 h dark were alternated with 12 h light. Only 7% migrated when light was constant for 24 h. Vertical migration of M. incognita juveniles was studied at 14, 16, 18, 20, and 22 C. The migration of M. incognita juveniles begins at about 18 C and reaches its maximum at 22 C. The migration of M. hapla and M. incognita juveniles were compared at 14, 18, and 22 C. Juveniles of M. hapla were able to migrate at a lower temperature than those of M. incognita. With M. hapla, there was no significant difference in migration between 18 and 22 C. PMID- 19300749 TI - Association of Macroposthonia xenoplax and Scutellonema brachyurum with the Peach Tree Short Life Syndrome. AB - High populations of both Scutellonema brachyurum and Macroposthonia xenoplax were detected in the root zones of peach trees growing at sites where peach tree short life occurred. Both nematodes responded to changes in tree health, with the highest numbers detected under the most vigorous trees; both populations declined as tree vigor declined. Few of either species were detected 6 months after trees died. The nematode populations were inversely correlated with each other. More M. xenoplax were recovered during cool, wet weather, but moisture seems more important than temperature in influencing population levels. Most S. brachyurum were recovered during hot, dry weather. PMID- 19300750 TI - A Laboratory Technique for Culturing Filipievimermis leipsandra, a Nematode Parasite of Diabrotica balteata Larvae (Insecta: Coleoptera). PMID- 19300751 TI - Comparative Susceptibility of Larval Mosquitoes Exposed Separately by Instar or in Mixed Populations to the Nematode Romanomermis culicivorax. PMID- 19300752 TI - Effects of Culture Age on the Shipment Survival of the Mosquito Parasite Romanomermis culicivorax. PMID- 19300753 TI - Observations on the Cuticle Ultrastructure of Meloidogyne hapla Males. PMID- 19300754 TI - A technique for establishing microplots in the field. PMID- 19300755 TI - Effect of gamma-Radiation on Dauer Larvae of Caenorhabditis elegans. PMID- 19300756 TI - Status and future of entomogenous nematodes: symposium introduction. PMID- 19300757 TI - Biological control potential of neoaplectanid nematodes. AB - The neoaplectanids are among the most studied of all entomogenous nematodes. Because these nematodes kill their insect hosts, they are regarded as having excellent potential as biological control agents. While the host specificity of most entontogenous nematodes tends to limit their potential usefulness, the broad host range and high virulence of neoaplectanids make them attractive candidates for industrial development. Also, recent development of economical mass rearing procedures appears to make production on a commercial basis feasible. Infective stages may be stored for years trader various laboratory conditions. Although entomogenous nematodes, as parasites, are exempt from govermnent registration requirements, the mutualistic association of neoaplectanid nematodes with a bacterium will likely necessitate a detailed safety evaluation. Studies conducted to date indicate a lack of pathogenicity to mammals. Field trial success has been limited by the intolerance of infective stages to mffavorable environmental conditions, particularly low moisture. Applications against pests on exposed plant foliage have been especially disappointing. More encouraging anti consistent results have been obtained in more favorable environments, including soil and aquatic habitats, but the most promising treatment sites ntay be cryptic habitats where infective stages are shehered from environmental extremes. Cryptic habitats also exploit the ability of neoaplectanids to actively seek out hosts in recessed places where conventional insecticide applications are intpractical. PMID- 19300758 TI - Mermithid parasitism of black flies (Diptera: simuliidae). AB - Mermithid nematodes are common parasites of black flies and play a significant role in the natural regulation of these medically important insects. Infection levels tend to he moderate and perennial, with epizootics rare and highly localized. Mermithid parasitism almost invariably results in the death of the black fly, and thus considerable attention has focused on the potential of these nematodes as biocontrol agents. Early instar black fly larvae appear most susceptible to infection, and integumental penetration hy mermithid preparasites is the only known mode of entry. Postparasitic nematodes typically emerge before host pupation. However, carryover of parasitism into adult simuliids is an important mechanism for local dispersal and recolonization of upstream areas. Following emergence, the mermithids molt to the adult stage. Copulation ensues, the females then laying eggs which eventually give rise to the next generation of infective preparasites. The number of described species is conservatively estimated at 35-40, with most species within the genera Mesomermis, Gastromermis, and Isomermis. The taxonomy of this group of mermithids is a challenging and little explored area. Host-specificity statements, therefore, must be made cautiously because of these systematic problems and others within the Simuliidae. In most instances, temporal and spatial factors limit the host range of these mermithids among simuliid species. Differential susceptibilities anmng larvae concurrently present within the same microhabitat probably reflect varying degrees of host attractiveness and behavioral-physiological resistance. Effects of parasitism on the host may include prevention of metamorphosis, sterility, intersexual development, and behavior modification. Evaluation of the technical feasibility of mermithid control of black flies has been stymied by the limitations of current inoculum-production technology. Continued advances in in vivo and in vitro culture methods are required to accelerate the research process. PMID- 19300759 TI - Biological control of mosquitoes with mermithids. AB - Mermithid nematodes parasitizing mosquitoes have substantial potential for vector control. Studies on the physiological ecology of Romanomermis culicivorax have defined some of the general requirements of mermithid nematodes and produced general guidelines for the experimental release of mermithids in biological control. Experimental field studies have established the biological control potential of R. culicivorax, but further development and ulilization of this parasite will require a substantial commitlnent of scientific man-years and ftmds. PMID- 19300760 TI - Mermithid parasites of agricultural pest insects. PMID- 19300761 TI - Mermithid Nematodes: Physiological Relationships with their Insect Hosts. AB - This paper assesses our state of knowledge of physiological processes involved in the relationships between insects and their mermithid nematode parasites. Three major components of the host-parasite relationship(s) are reviewed: effects of mermithids on host physiology, effects of host physiology on mermithids, and the physiology of the nematodes themselves. Mermithids induce an array of changes in host physiology, and the effects on host metabolism and endocrinology are discussed at some length. Few studies have been done to ascertain the effects of the host on the parasites from a physiological standpoint. Whereas host immunity mechanisms against mermithids have been described at the ultrastructural level, the physiological basis of such responses is not known. Mermithids are atypical nematodes, both structurally and physiologically. In the absence of a functional gut, nutrients are absorbed across the outer cuticle and stored in a trophosome. The transcuticular mode of feeding, storage within the trophosome, and metabolism of storage products are discussed. The usefulness of physiological information toward expediting in vitro culture of these nematodes is discussed, and problems that need to be addressed are defined. PMID- 19300762 TI - Mermithid nematodes: in vitro culture attempts. AB - Few attempts at in vitro culture of mermithids have been undertaken. The various methods used to initiate cultures are described. The capacity of a range of media to promote growth and development of the nematodes has been evaluated and current approaches to in vitro outlined. PMID- 19300763 TI - Commercial development and future prospects for entomogenous nematodes. AB - Although entomogenous nenmtodes generally have many of the attributes of the ideal biocontrol agent, many of these attributes make the nematodes less than desirable for commercial production. Environmental limitations, lack of patent protection, "shelf life," shipping problems, and the need for users to receive specialized training are factors that have discouraged the involvement of larger companies. The future of these nematodes as commercially available biocontrol products appears to lie with the smaller "cottage industries" or with government subsidized production. Problems encountered with attempts to produce commercially the mosquito parasite Romanomermis culicivorax are discussed. PMID- 19300764 TI - Effects of Host Diet on Romanomermis culicivorax, a Mermithid Parasite of Mosquitoes. AB - When larval mosquitoes (Aedes aegypti) infected with the mermithid nematode Romanomermis culicivorax were fed on a diet low in quantity or protein content or both, the number of postparasites which emerged from the hosts decreased and host mortality increased marginally. Parasitic development was prolonged and became asynchronous in nutritionally deprived hosts. Nematodes emerged from insects infected by more than one nematode before the remaining juveniles comprising such infections had completed parasitic development; this resulted in substantial reductions in postparasite nttmbers. Host development was retarded by low protein and/or reduced diets. Postparasites emerging from second and third instars were reduced in size and in the amount of stored nutriment compared to those recovered from hosts fed on a high protein diet ad libitum. A greater proportion of the rnermithids developed into males in hosts fed on reduced diets but not in hosts fed on low protein diets. PMID- 19300765 TI - Susceptibility of Various Species of Lepidopterous Pupae to the Entomogenous Nematode Neoaplectana carpocapsae. AB - The susceptibility of certain species of lepidopterous pupae occurring in different ecological situations to the entomogenous nematode, Neoaplectana carpocapsae, was tested. Soil- or litter-pupating lepidopterous insects were not highly susceptible to N. carpocapsae. The most susceptible insect pupating in the soil was Spodoptera exigua with 63% pupal mortality, while Harrisinia brillians, which pupates in litter, had 55% mortality. Other soil- and litter-pupating insects had mortalities of less than 25%. Some insect species that pupate above ground were highly susceptible (> 84% mortality) to N. carpocapsae infection. PMID- 19300766 TI - Dispersal and Infectivity of the Entomogenous Nematode, Neoaplectana carpocapsae Weiser (Rhabditida: Steinernematidae), in Sand. AB - Laboratory tests determined the lateral and vertical dispersal patterns of Neoaplectana carpocapsae in sand. In the vertical tests, placement of infective juveniles 15 cm below the sand's surface resulted in the majority (77%) being recovered above the point of placement after 48 h. Placement of the nematodes on the sand's surface resulted in the majority (90.4%) remaining within 1 cm of the sand's surface. Placement of nematodes at depths of 2.5 cm and 5.0 cm below the sand's surface also resulted in little nematode dispersal. However, vertical hioassay tests showed that juvenile nematodes placed on the sand's surface dispersed 12 cm down to infect 67% of the Galleria mellonella pupae placed at the depth. Conversely, when nematodes were placed 11 cm below the insect pupae no infection was observed, but 53% infection occurred when nematodes were 7 cm below the site of the insect pupae. In lateral dispersal, 87% of the nematodes rentained within 2 cm of the placement site, although 0.5% were recovered at 12 14 cm away from the point of placement. Lateral bioassay tests indicated that the nematodes were capable of infecting 90, 35, and 5% of the G. mellonella pupae at 7 cm, 10 cm, and 14 cm from the point of placement, respectively. PMID- 19300767 TI - An Algorithm for Fitting Seinhorst Curves to the Relationship between Plant Growth and Preplant Nematode Densities. PMID- 19300768 TI - Sample optimization for five plant-parasitic nematodes in an alfalfa field. AB - A data base representing nematode counts and soil weight from 1,936 individual soil cores taken from a 7-ha alfalfa field was used to investigate sample optimization for five plant-parasitic nematodes: Meloidogyne arenaria, Pratylenchus minyus, Merlinius brevidens, Helicotylenchus digonicus, and Paratrichodorus minor. Sample plans were evaluated by the accuracy and reliability of their estimation of the population and by the cost of collecting, processing, and counting the samples. Interactive FORTRAN programs were constructed to simulate four collecting patterns: random; division of the field into square sub-units (cells); and division of the field into rectangular sub traits (strips) running in two directions. Depending on the pattern, sample numbers varied from 1 to 25 with each sample representing from 1 to 50 cores. Each pattern, sample, and core combination was replicated 50 times. Strip stratification north/south was the most optimal sampling pattern in this field because it isolated a streak of fine-textured soil. The mathematical optimmn was not found because of data range limitations. When practical economic time constraints (5 hr to collect, process, and count nematode samples) are placed on the optimization process, all species estimates deviate no more than 25 % from the true mean. If accuracy constraints are placed on the process (no more than 15% deviation from true field mean), all species except Merlinius required less than 5 hr to complete the sample process. PMID- 19300769 TI - Plant-parasitic Nematodes in Loess Toposequences Planted with Corn. AB - Populations of plant-parasitic nematodes in an Iowa cornfield were studied along north- and west-facing toposequences. Samples were collected monthly during the growing season. The greatest biomass for Xiphinema americanum occurred at the footslope on the north face. Paratylenchus microdorus had its greatest biomass at the summit position, generally more in the west- than in the north-facing slope. Pratylenchus spp. in the roots peaked at the toeslope in the north-facing slope, but at the footslope in the west-facing slope. Helicotylenchus pseudorobustus peaked at the backslope and the toeslope along the north- and west-facing slopes, respectively. Diversity, as computed for each plot by the Shannon-Weiner diversity index, was highest at the backslope in both toposequences. PMID- 19300770 TI - Enhancement of Cylindrocladium crotalariae Root Rot by Meloidogyne arenaria (Race 2) on a Peanut Cultivar Resistant to Both Pathogens. AB - Two populations of Meloidogyne arenaria (race 2, incompatible on peanut) enhanced development of Cylindrocladium black rot (CBR) on CBR-resistant peanut cv. NC 3033 in greenhouse factorial experiments. Nematode populations 256 and 486 (0, 10(3), 10 eggs per 15-cm pot) were tested in all combinations with Cylindrocladium crotalariae (0, 0.5, 5, 50 microsclerotia per cm(3) of soil). Root-rot index increased in the presence of either population. Positions but not slope values of inoculum density-disease curves were changed by both populations, indicating increased efficiency of microsclerotia when peanuts were grown in the presence of these nematodes. Although little or no reproduction occurred with either nematode population on NC 3033, larvae of 256 and 486 penetrated roots. Meloidogyne arenaria 486 did not induce root galls and was not snccessful in establishing feeding sites. Meloidogyne arenaria 256 produced a few very small eliptical galls and had a range of success in establishing a feeding site, varying from no giant cell development to large giant cell with production of a few eggs. PMID- 19300771 TI - Damage and Reproductive Potentials of Pratylenchus brachyurus and P. penetrans on soybean. AB - Damage and reproductive potentials of Pratylenchus brachyurus and P. penetrans on soybean, Glycine max, cvs. Essex, Forrest, and Lee 68, were determined in microplot tests. Cultivar Essex was generally tolerant to P. brachyurus. Yield of Forrest was suppressed linearly with increasing P(i)'s in the sandy soil (r = 0.92) and loamy sand soil (r = -0.99). Low to moderate P(i)'s in the sandy clay loam gave an increase in yields as compared to plants without nematodes. Yield was not affected by this nematode in muck. Lee 68 was very sensitive to P. penetrans in microplots. Yield vs. P(i) was fitted by a quadratic model (r = 0.82) with yield decreasing sharply as P(i)'s were increased. The reproduction of both species decreased with increases in P(i). Lee 68 was a good host for P. penetrans, whereas Essex and Forrest were fair to poor hosts for P. brachyurus. PMID- 19300772 TI - Influence of Soil Temperature and pH on Pratylenchus Penetrans and P. crenatus in Alfalfa and Timothy. AB - Numbers of Pratylenchus penetrans in alfalfa and timothy, and to a lesser extent P. crenatus in timothy, increased substantially as temperature increased from about l0 C to 30 C. However, P. crenatus in alfalfa decreased in number as temperature increased. Mobility of P. crenatus in vertical soil columns decreased as temperature increased from 9.5 C to 28.5 C. Raising the soil pH from 5.0 to 6.9 in which alfalfa was grown increased the numbers of P. penetrans and greatly reduced the numbers of P. crenatus. The numbers of both nematode species in timothy were reduced significantly as soil pH was increased. The optimum soil pH for movement of P. penetrans was 6.0. Pratylenchus crenatus moved equally well over a range of pH 5.0 to 7.0. PMID- 19300773 TI - Effects of Soil Temperature and Planting Date of Wheat on Meloidogyne incognita Reproduction, Soil Populations, and Grain Yield. AB - Wheat cultivars Anza and Produra grown in winter in California were planted in Meloidogyne incognita infested and noninfested sandy loam plots in October (soil temperature 21 C) and November (soil temperature 16 C) of 1979. Meloidogyne incognita penetrated roots of mid-October planted Ataza (427 juveniles/g root), developed into adult females by January, and produced 75 eggs/g root by harvest in April. Penetration and development did not occur in late plantings. Anza seedlings grown in infested soil in pots buried in field soil in early spring were not invaded until soil temperature exceeded 18 C. Meloidogyne incognita juveniles can migrate through soil and penetrate roots at temperatures above 18 C (activity threshold), however development can occur at lower temperatures. Grain yields were not significantly different between nematode infested (3,390 kg/ha) and noninfested (2,988 kg/ha) plots. Winter decline of eggs and juveniles in two late plantings anti in fallow soil were 69, 72, and 77%, respectively, but egg and juvenile decline was only 40% in the early Anza plots that supported nematode reproduction in the spring. Delay of planting date until soil temperature is below 18 C is suggested to maximize the use of wheat in rotation as a nematode pest management cultural tactic for suppressing root-knot nematodes. PMID- 19300774 TI - The Development and Influence of Meloidogyne incognita and M. javanica on Wheat. AB - The effects of soil temperature and initial inoculum density (Pi) of Meloidogyne incognito and M. javanica on growth of wheat (Triticum aestivum cv. Anza) and nematode reproduction were studied in controlled temperature baths in the glasshouse. Nematode reproduction was directly proportional to temperature between 14 and 30 C for M. incognita and between 18 and 26 C for M. javanica. Reproduction rates (Pf/Pi, where Pf = final number of eggs) for Pi's of 3,000, 9,000, and 30,000 eggs/plant were greatest at each temperature when Pi = 3,000. Maximum M. incognita reproduction rate (Pf/Pi = 51.12) was at 30 C. At 26 C, M. javanica reproduction (Pf/Pi = 14.82, 9.02, and 4.23 for Pi = 3,000, 9,000, and 30,000, respectively) was about half that of M. incognita when Pi = 3,000 or 9,000 but similar when Pi = 30,000. Reproduction of both species was depressed between 14 and 18 C. Shoot and root growth and head numbers were inversely related to soil temperature between 14 and 30 C but were not affected by the Pi of M. incognita when 7 d old seedlings were inoculated. When newly germinated seedlings were inoculated with M. incognita or M. javanica, the Pi did not affect shoot and root fresh weights, shoot/root ratio, and tillering, but it did reduce root dry weight (M. javanica at 26 C) and increase shoot dry weight (M. incognita at 18-22 C). The optimum temperature range is lower for wheat growth than for nematode reproduction. Wheat cv. Anza is a good host for M. incognita and M. javanica, but it is tolerant to both species. PMID- 19300775 TI - Resistance of Trisomic and Diploid Hybrids of Beta vulgaris and B. procumbens to the Sugarbeet Nematode, Heterodera schachtiis Arnold. AB - Trisomic and diploid hybrids of sugarbeet (Beta vulgaris L.) X wild beet (B. procumbens Chr. Sin.) inherited the gene for resistance to Heterodera schachtii Schm. from B. procumbens. The hybrids showed partial resistance to H. schachtii, manifested in failure of larvae to reach maturity. Although significantly greater numbers of female nematodes developed on plants inoculated with populations from the Netherlands or Italy than on plants inoculated with a population from the Salinas Valley, California. the totals for all populations on resistant plants were small. Greater numbers of males than females developed on root-slice cultures of resistant hyhrids when compared to a susceptible cultivar. PMID- 19300776 TI - Scanning Electron Microscope Study of the Root-knot Nematode (Meloidogyne incognita) on Tomato Root. AB - This study examines the types of structural information that can be gained by utilizing the scanning electron microscope (SEM) and a cryofracture technique to examine the host-parasite interaction. Roots of tomato, Lycopersicon esculentum cv. Marglobe, were cultured aseptically and inoculated with the root-knot nematode, Meloidogyne incognita. Twenty-four hours to four weeks after inoculation, developing galls were removed from the cultures and processed for SEM observation. The cryofracture technique was used to reveal internal structural features within the developing galls. The results illustrate structural details concerning penetration of the roots, differentiation of syncytia, and development of the nematodes beginning with the second-stage larvae and ending with adult egg-laying females. PMID- 19300777 TI - Resistance and Resistant Reaction of Gossypium arboreum to the Reniform, Nematode, Rotylenchulus reniformis. AB - Gossypium arboreum 'Nanking CB 1402' possessed a high level of resistance to Rotylenchulus reniformis. Within 16 h, the nematode penetrated roots of resistant and susceptible cottons equally. After 36 h, significantly fewer nematodes were found in resistant roots. Larvae fed in either an endodermal or pericyclic cell and had no specificity for root tissue of a particular age. In roots of resistant G. arboreum '1402,' wall breakdown of pericyclic cells was evident after 3 d, endodermal and cortical cells collapsed, and the hypertrophied pericyclic cells disintegrated within 12 d. Cell walls immediately adjacent to the nematode's head were thickened and more safranin positive in resistant than in susceptible cotton cultivars. Several other cultivars of G. arboreum were also resistant to R. reniformis, based on nematode fecundity and percent egg reduction. PMID- 19300778 TI - The Effect of Meloidogyne incognita and Tissue Wounding on Severity of Seedling Disease of Cotton Caused by Rhizoctonia solani. PMID- 19300779 TI - Observations on the Morphology of the Pine Wood Nematode, Bursaphelenchus xylophilus. AB - Scans of slash pine wood chips infested with pine wood nematodes showed coiled aggregates of 5-10 nematodes in the axial resin canals, and 1-2 nematodes folded in the radial resin canals. Observations of the pine wood nematode by SEM showed the head to be offset with six labial lobes. A smaller circle of six inner lobes surrounded the stoma. A single labial sensillum was observed on each of the two subventral and two subdorsal lobes. The two opposite lobes, without sensilla, bore a pore-like amphid. The female vulva flap was observed to be a pouch. In addition to two pairs of copulatory papillae previously described for the male, three additional papillae were observed: an additional post-anal pair, and a single papilla anterior to the cloaca. Spicules were fused ventrally. Distal spicule ends terminated in two fused half discs. Caudal alae were angular at the tail terminus, but culed inward at the corners to appear oval. Lateral fields had four lateral striations. PMID- 19300780 TI - On the Taxonomy and Morphology of the Pine Wood Nematode, Bursaphelenchus xylophilus (Steiner &Buhrer 1934) Nickle 1970. AB - During the past 3 yr, nematologists in the United States have found specimens of Bursaphelenchus sp. in the wood of dead and dying pine trees. This nematode-host association resembles a similar interaction reported from Japan where pine trees are being killed by the pine wood nematode. This taxonomic research was conducted to determine if the Japanese pine wood nematode and similar populations in the United States are of the same species. Based upon typical morphological characters of original specimens of Bursaphelenchus xylophilus (Steiner and Buhrer 1934) Nickle 1970 that were rediscovered in the USDA Nematode Collection and genetic crosses among the Japanese and American nematode populations, it was concluded that they are all the same species, B. xylophilus. PMID- 19300781 TI - Description of Meloidogyne nataliei n. sp. (Nematoda: Meloidogynidae) from Grape (Vitis labrusca) in Michigan, with SEM Observations. AB - Meloidogyne nataliei n. sp. is described and illustrated from grape (Vitis labrusca) in a declining vineyard at Mattawan, Michigan, USA. Infected grape roots exhibit no hyperplastic symptoms. Females protrude from the roots and are surrounded by a massive egg sac containing many eggs. This new species is distinguishable from other species of the genus especially by its large, striking perineal pattern with, usually, two ropelike separated striae on each side extending laterally from the vulval and anal areas. Among other diagnostic characters are the location of the female excretory pore adjacent to or near the base of the head and the heavy larval stylet averaging about 22 mum in length. Examination of males, females, and larvae with the scanning electron microscope confirmed observations made by optical microscopy and revealed diagnostic and other structures in greater detail. Of particular significance was the nature of the male head, with a massive circular labial disc on which is located a rectangular structure surrounding the oral opening, and the six distinct lips appearing as a rosette in en face view. The known distribution of this new species is presently limited to its original location in Michigan. PMID- 19300782 TI - Brachydorus swarupi sp. n. (Nematoda: Dolichodorinae) from Soil about Roots of Arecanut Palm in Kerala State, India. AB - Brachydorus swarupi sp. n. is described from soil about roots of arecanut palm in Kerala State, India. It is the second species to be described in Brachydorus De Guiran and Germani 1968 and differs from B. tenuis, the type species, by its greater length (1.52-2.34 mm for B. swarupi vs. 1.03-1.32 mm for B. tenuis); longer and more delicate stylet (26-35 mum for B. swarupi vs. 20-23 mum for B. tenuis); head shape with concave protrusion at oral aperture (simple rounded in B. tenuis); shorter isthmus and larger posterior bulb in B. swarupi; shorter tail (in B. tenuis c = 9.7 [8.6-11.5] in female, c = 42.3 [33-48] in male); larger spicules and gubernaculum (22-39 mum and 9 -1 2 mum, respectively, for B. tenuis); and phasmids near posterior connection of caudalalae and tail (almost central on caudalalae of B. tenuis). The relationship of Brachydorus to Dolichodorus is discussed. PMID- 19300783 TI - Nematicide treatments of turfgrass seed. AB - Seed treatments of improved Kentucky bluegrass and fescue cultivars with carbofuran, oxamyl, and phenamiphos dissolved in acetone reduced seedling emergence, but treatments were not extremely phytotoxic. Phenamiphos was the most toxic, particularly at the 5,000 mug/ml concentration. Fresh weight of grass clippings 35 d following planting generally was greater in treatments than in controls except for the 5,000 mug/ml phenamiphos treatments on certain cultivars. All nematicide seed treatments reduced the number of Pratylenchus penetrans subsequently recovered from Pennlawn creeping red fescue roots 4-5 wk after treatment. The infusion of nematicides into grass seed with organic solvents appears to be an effective means of reducing nematode damage to turfgrass seedling with little environmental hazard. PMID- 19300784 TI - Influence of volcanic ash on infectivity and reproduction of two species of meloidogyne. AB - Mount St. Helens volcanic ash was incorporated into a loamy sand greenhouse soil mix to produce concentrations of 0, 0.5, 1.0, 2.0, 4.0, 8.0, 25, 50 and 100% ash. Chemical and physical properties of the various mixtures were determined. Three experiments were conducted in a greenhouse to determine if volcanic ash had any influence on root-knot nematode survival and infectivity. Tomato, Lycoperscion esculentum, seedlings cv. Columbia, susceptible to Meloidogyne hapla and M. chitwoodi were planted into pots of the soil-ash concentrations and infested with one of the two nematode species. Tomato seedlings were harvested 30, 50 and 60 days later and the roots examined for nematode infection and reproduction. Ash incorporation had no deliterious effect on root-knot nematodes in any of the experiments reported here. Nematode infection and reproduction on tomato were not affected at any ash concentration. PMID- 19300785 TI - Beta-Glycosidases from Meloidogyne incognita and M. javanica. PMID- 19300786 TI - A versatile nematode water bath apparatus. PMID- 19300787 TI - Foliar Spray Effects of Selected Amino Acids on Sunflower Infected with Meloidogyne incognita. PMID- 19300788 TI - Susceptibility of Pupae of Two Cocoon-forming Lepidopterous Species to the Entomogenous Nematode, Neoaplectana carpocapsae (Rhabditida: Steinernematidae). PMID- 19300789 TI - Distribution of Pheromermis pachysoma (Mermithidae) Determined by Paratenic Invertebrate Hosts. PMID- 19300790 TI - Mass Production of the Entomogenous Nematode Heterorhabditis heliothidis (Nematoda: Heterorhabditidae) on Artificial Media. AB - Heterorhabditis heliothidis is reared monoxenically on an artificial medium consisting of commercially available nutrient broth, yeast extract, and vegetable oil. These components are cooked with flour and coated onto polyether polyurethane sponge, autoclaved, inoculated with a suspension of the bacterial symbiont of the nematode, and incubated at 25 C for 3 d. The bacterial garden on sponge provides an excellent rearing medium. Up to 10 million infective juveniles are produced per 250 ml rearing flask in one month. PMID- 19300791 TI - Gaseous Requirements for Postparasitic Development of Romanomermis culicivorax. AB - The development of postparasitic stages of Romanomermis culicivorax was studied under various concentrations of oxygen and carhon dioxide. The nematode developed poorly if only nitrogen was supplied; only one-third molted and all died eventually. In the presence of 5% CO - 95% N, development was normal; most nematodes molted and oviposited with respective mean developmental times of 32 and 50 d. Addition of 0.2% O stimulated development; molting and oviposition commenced at days 18 and 41, respectively. There was an additional stimulation of development by increasing amounts of O up to 1%, but concentrations greater than 1% produced no additional stimulation. Carbon dioxide was required for development after exsheathment under anaerobic conditions or O concentrations less than 1%. Oxygen or CO were not required for embryological development or egg hatch. It is suggested that post-parasitic stages function as facultative anaerobes. PMID- 19300792 TI - On the transport of nematodes by the wind. AB - The possible effectiveness of atmospheric transport of nematode forms (dry larvae or eggs) as a means for introducing new species to a given environment is examined. Given the measured sedimentation velocities for a range of forms (0.1 >/= Ws >/= 0.6 mps), the necessary conditions on the wind speed required for natural erosion are defined. With these results scenarios for lofting, transport, and diffusion of these forms are examined using relevant gaussian plume models. Results indicate that on rare occasions individuals can be deposited up to 40 km from their original location. Redepositions up to 5 km per erosion event should be fairly common occurances when dry loose soil conditions or dry tillage operations combine with optimal atmospheric conditions and the presence of significant numbers of nematodes at the surface. PMID- 19300793 TI - Effect of Soil Temperature on the Pathogenicity and Reproduction of Meloidogyne chitwoodi and M. hapla on Russet Burbank potato. AB - Meloidogyne chitwoodi and M. hapla were pathogenic to both roots and tubers of Russet Burbank potato. Both species affected root growth at 15, 20, and 25 C, but not 30 C. Meloidogyne chitwoodi reprotluced best at 15, 20, and 25 C and M. hapla at 25 and 30 C. Reproduction of M. chitwoodi was reduced at 30 C; reproduction of M. hapla was reduced at 15 C and less at 20 C. The reproductive potential of M. chitwoodi was higher than that of M. hapla at 15, 20, and 25 C. M. hapla reproduced better at 30 C than did M. chitwoodi. M. chitwoodi infected potato tubers in higher numbers than did M. hapla. PMID- 19300794 TI - Histopathology of Meloidogyne chitwoodi (Golden et al.) on Russett Burbank Potato. AB - Pathogenesis of M. chitwoodi associated with potato (Solanum tuberosura cv. Russet Burbank) followed a pattern characteristic of root-knot nematodes. Giant ceils developed in the phloem tissues of roots, stolons, and tubers and appeared to arise by hypertrophy and karyokinesis rather than cellular fusion. Gall formation was a function of parasite density and developed by hypertrophy of cortical cells. Brownish lesions which are symptomatic of tuber infection resulted from lignification of cortical cell walls in contact with egg matrix. PMID- 19300795 TI - Characterization of Citrus Rootstock Responses to Tylenchulus semipenetrans (Cobb). AB - Citrus rootstocks which significantly limited the reproduction of Tylenchulus semipenetrans (Cobb) "Citrus" and "Poncirus" biotypes responded to infection by producing a hypersensitive-type response in the root hypodermis, wound periderm and/or cavities in the root cortex, and/or abnormal vacuoles in nurse cell cytoplasm. Rootstocks which limited nematode reproduction also had significantly fewer nematodes in the rhizoplane within 8 d of inoculation than did rootstocks which did not limit reproduction. Germplasm sources of the cellular responses which limited citrus nematode reproduction were identified. PMID- 19300796 TI - Three new species of heteroderoidea (nematoda) from the aleutian islands. AB - Three new species of Heteroderoidea are described from Adak and Amchitka Islands in the Aleutian chain. Second-stage juveniles of Thecavermiculatus crassicrustata, n. sp., differ from those of T. gracililancea Robbins by having longer stylets (40-45 mum vs 19-22 mum). The female of T. crassicrustata has a longer neck, a more posterior excretory pore, and lacks a posterior protuberance. Meloidodera eurytyla, n. sp., differs from other Meloidodera spp. in that second stage juveniles have longer stylets (32-35 mum) and much more massive stylet knobs, while males have a longitudinally striated basal head annule. Meloidogyne subarctica, n. sp., can be separated from other Meloidogyne spp. by combinations of the following characteristics: perineal pattern with large oval areas in the tail region devoid of striae, arch with few unbroken striae; female excretory pore 1.5-2.5 x the stylet length from the anterior end; haploid chromosome number = 18; the spermatheca filled with sperm; stylet length of second-stage juveniles 13.5-15.4 mum. PMID- 19300797 TI - Identification of meloidogyne species on the basis of head shape and, stylet morphology of the male. AB - Head shape and stylet morphology of males of 90 populations of M. arenaria, M. hapla, M. incognita, and M. javanica from geographic regions of the world were compared by light microscopy (LM). In addition, stylets of one population each of M. arenaria, M. incognita, and M. javanica and three different chromosomal forms of M. hapla race A and two of race B were excised and examined with a scanning electron microscope (SEM). Differences among species occurred in both head and stylet morphology. Head morphology differed in size and shape of the head cap, annulation of the head region, and width of the head region relative to the first body annule. Differences in stylets occurred in size and shape of the cone, shaft, and knobs. All populations of M. hapla, except one, had similar head morphology, but stylet morphology was different between cytological races A and B. Populations of M. javanica varied with respect to the presence of head annulations. Head shape and stylet morphology of males are recommended as additional characters useful in the identification of root-knot nematodes. PMID- 19300798 TI - Redescription and Lectotype Designation of Tylenchorhynchus cylindricus Cobb, 1913. AB - Tylenchorhynchus cylindricus is redescribed and illustrated from N. A. Cobb's original specimens collected in 1910. In 1955 M. W. Allen established a neotype from specimens collected near Cathedral City, California. Recently Cobb's original sketches, line drawings, and balsam slides were rediscovered and examined. The specimens collected by Cobb were compared with the neotype established by Allen and with other collections of nominal T. cylindricus. Differences in morphology of the Cathedral City (Allen) and Los Patos (Cobb) populations were observed. Collections of males and females from Cathedral City, California; Mosida, Utah; and Kings County, California; were similar to each other except for some variation in female tail shape. Females in Cobb's collection and in a collection from a beach near Ensenada, Mexico, were similar to each other but differed morphologically from other collections. We consider all collections to represent a range of variation within the species. A lectotype and an allolectotype were selected to establish the taxonomic base for the genus. A ruling has been requested from the International Commission of Zoological Nomenclature on the disposition of the neotype. PMID- 19300799 TI - Effects of Oxygen and Temperature on the Activity and. Survival of Nothanguina phyllobia. AB - The effects of oxygen and temperature on the activity and survival of infective forth-stage juveniles of Nothanguina phyllobia Thorne were examined in aqueous suspension. Rate of movement was not affected by a wide range of O concentration (0.8-8.6 ppm). Activity decreased below 0.8 ppm 0 2, and at 0.15 ppm O nematodes became motionless. Activity increased as a linear function of temperature up to a thermal optimum of 24 C; beyond 24 C activity decreased. Survival was greatly prolonged at low temperature. At 23 C, 50% mortality occurred within 7 d, whereas at 4 C, 70% survived after 98 d. PMID- 19300800 TI - A Soil-free System for Assaying Nematicidal Activity of Chemicals. AB - A biological assay system for studying the nematicidal activity of chemicals has been devised using a model consisting of cucumber (Cucumis sativus L. cv. Long Marketer) seedlings growing in the diSPo(R) growth-pouch apparatus. Meloidogyne incognita was used as the test organism. The response was quantified in terms of the numbers of galls produced. Statistical procedures were applied to estimate the ED(50) values of currently available nematicides. This system permits accurate quantification of galling and requires much less space and effort than the currently used methods. PMID- 19300802 TI - Herbicide effects in nematode diseases. PMID- 19300801 TI - Distribution and population dynamics of nematodes in a rice field and pasture in India. AB - Ecological studies on soil nematodes were made in a tropical rice field and pasture. Parasitic species were more diversified in the pasture than in the rice field. Eighty-six and sixty percent of total nematodes occurred in the top 10 cm in rice field and pasture, respectively. Nematodes were not randomly or uniformly dispersed but aggregated. Parasitic forms were most abundant and correlated with root biomass in the 0-15-cm soil layer, the greatest number usually occurring at the 10-15-cm depth at both sites. In summer, however, they were densest at the 15 30-cm depth. Microbivores were most frequent in the top 5 cm of both sites. Micellaneous feeders (food sources uncertain) usually occurred in highest densities at the 15-30-cm depth. Predators showed no distinct depth preference. Temperature and moisture of the soil apparently played an important role in regulating nematode population. Peak densities of 31.3 x 10/m(2) and 21.6 x 10/m(2) at a 30-cm depth occurred in January, while minimum densities of 5.0-5.3 x 10/m(2) and 4.1 x 10/m(2) occurred in July-October and April in rice field and pasture, respectively. Monthly mean biomass of nematodes was 23.8 +/- 4.5 mg/m(2) in rice field and 11.5 +/- 1.5 mg/m(2) in pasture. PMID- 19300803 TI - Parasitism of Walnut, Juglans regia, by Cacopaurus pestis. PMID- 19300804 TI - Pathogenicity of the Columbia Root-knot Nematode (Meloidogyne chitwoodi) on Wheat, Corn, Oat, and Barley. PMID- 19300805 TI - Hydroxytetraphenylenes, a new type of self-assembling building block and chiral catalyst. AB - Tetraphenylenes are structurally exceptional molecules with rigid conformations so that substituents of these compounds demonstrate highly directional dispositions. As such, these molecules are potential candidates for employment as organocatalysts or ligands in asymmetric synthesis. Careful design and manipulation of the structural features of tetraphenylenes should also lead to the formation of supramolecular helical scaffolds or other supermolecules with fascinating arrangements. We would like to present here a brief summary on the chemistry of some hydroxytetraphenylenes that were synthesized in our laboratories in Shanghai and Hong Kong. This article will serve as a preview of the chemistry of the tetraphenylene family that hopefully has the potential to emerge. PMID- 19300806 TI - Design and synthesis of phosphole-based pi systems for novel organic materials. AB - Phospholes are known to exhibit characteristic optical and electrochemical properties derived from the phosphorus-bridged 1,3-dienic pi system. Particular interest has recently been paid to their pi-conjugated derivatives, such as non fused phospholes, dibenzo[b,d]phospholes, benzo[b]phospholes, benzo[c]phospholes and related compounds. This perspective focuses on recent advances in the synthesis of these phosphole-based pi systems with representative examples that would give valuable guidelines for designing phosphole-containing opto electrochemical materials. PMID- 19300807 TI - Trifunctional organocatalyst-promoted counterion catalysis for fast and enantioselective aza-Morita-Baylis-Hillman reactions at ambient temperature. AB - Fast and enantioselective aza-Morita-Baylis-Hillman reactions between electron deficient or electron-rich aromatic N-tosyl imines and methyl vinyl ketone were achieved at ambient temperature using asymmetric counterion-directed catalysis promoted by trifunctional organocatalysts with a Bronsted base as the activity switch after protonation with benzoic acid. PMID- 19300808 TI - Silver-catalysed Doyle-Kirmse reaction of allyl and propargyl sulfides. AB - The silver-catalysed Doyle-Kirmse reaction of propargyl and allyl sulfides with diazo compounds is disclosed. The carbon-carbon bond forming process proceeds with a range of substituents and functionality under mild conditions. PMID- 19300809 TI - Reaction of carbohydrates with Vilsmeier reagent: a tandem selective chloro O formylation of sugars. AB - A convenient and efficient method for selective replacement of the primary hydroxyl groups of sugars by chlorine with concomitant O-formylation, compatible with the presence of a variety of functional groups, has been developed using the Vilsmeier-Haack reaction. Sugars having free primary hydroxyl groups mostly afforded the chloro-O-formylated product while sugars devoid of primary hydroxyl groups yielded only O-formylated products. PMID- 19300810 TI - Highly stereoselective synthesis of aminoglycosides via rhodium-catalyzed and substrate-controlled aziridination of glycals. AB - The flexible installations of a sulfamate ester on a glycal scaffold at C3, C4, or C6 approaching alpha- or beta-aminoglycosides is communicated. A variety of glycal acceptors (O, S, and N) were applied, enhancing the utility of this method as an operationally simple protocol for the stereoselective synthesis of polyfunctionalized alpha- or beta- aminosaccharides. PMID- 19300811 TI - Three-dimensional bis(m-phenylene)-32-crown-10-based cryptand/paraquat catenanes. AB - Two three-dimensional catenanes have been successfully synthesized from a bis(m phenylene)-32-crown-10-based cryptand and paraquat derivatives in reasonable yields. PMID- 19300812 TI - Origins of enantioselectivity in the chiral Bronsted acid catalyzed hydrophosphonylation of imines. AB - The results of an experimental and ONIOM-based computational investigation of the mechanism and the origins of enantioselectivity in the asymmetric synthesis of alpha-amino phosphonates by an enantioselective hydrophosphonylation of imines catalyzed by chiral Bronsted acids are reported. It was found that the enantioselectivity observed in the enantioselective hydrophosphonylation of the imine with a benzothiazole moiety was poor. A detailed computational study with a two-layer ONIOM (B3LYP/6-31G(d)/AM1) method on the mechanism of the investigated reaction was carried out to explore the origins of the enantioselectivity. Calculations indicate that the investigated reaction is a two-step process involving proton-transfer and nucleophilic addition, which is the stereo controlling step. The investigated reaction prefers a di-coordination pathway to a mono-coordination pathway. The different enantioselectivities exhibited by three kinds of catalyst and two kinds of nucleophile were rationalized. Calculations indicate that si-facial attack is higher in energy than re-facial attack by only 0.1 kcal/mol, which accounts well for the low ee value observed in the enantioselective hydrophosphonylation of the imine with a benzothiazole moiety. The energy barrier for phosphonate-phosphite tautomerism catalyzed by chiral Bronsted acid in toluene is only 1.8 kcal/mol, which could explain why the investigated reaction can take place at room temperature. PMID- 19300813 TI - A red-shift colorimetric and fluorescent sensor for Cu2+ in aqueous solution: unsymmetrical 4,5-diaminonaphthalimide with N-H deprotonation induced by metal ions. AB - 4,5-Diamino-1,8-naphthalimide (DNP)-based chemosensor was designed and synthesized. Probe specifically recognized Cu(2+) ions in neutral aqueous solution. The capture of Cu(2+) by the receptor resulted in deprotonation of the secondary amine conjugated to the 1,8-naphthalimide chromophore, so that the electron-donation ability of the "N" atom would be greatly enhanced; thus probe showed a 50 nm red-shift in absorption (from 464 nm to 514 nm) and a large colorimetric response, it also exhibited an on-off fluorescent response. Specifically, is a pH-independent sensor in the range of 6.0 to 12.0 and can be used as a probe for Cu(2+) in strong basic conditions. These results opened up new possibilities for construction of red-shift colorimetric and fluorescent sensors. PMID- 19300814 TI - Synergistic activation of the Diels-Alder reaction by an organic catalyst and substituents: a computational study. AB - Density functional theory (DFT), using the hybrid functionals B3LYP and B2PLYP, has been employed to investigate the activation of the acrolein-butadiene Diels Alder reaction, mediated by a thiourea catalyst. Effects due to electron-donating groups (EDGs) on the diene, as well as electron-withdrawing groups (EWGs) on the dienophile, have also been studied. Organic catalysts such as thioureas are known to lower the activation energy through hydrogen-bonding to the carbonyl oxygen, in a way that mimics the oxyanion holes of hydrolytic enzymes. EDGs and EWGs were found to further activate the reaction, and the catalyst showed a synergistic behavior towards the EDGs. Polar solvents were found to reduce the overall activation energy, but also the relative catalytic effect of the thiourea, in accordance with experimental studies. The substituent-mediated reactions displayed more asynchronous transition structures with lower activation energy, which led us to investigate the possibility of an alternative two-step, Michael type route, similar to what has been found in macrophomate synthase. Although the concerted Diels-Alder route was found to be favored over the Michael route, the calculated activation energy difference is less than 1 kcal mol(-1), which suggests that the two mechanisms compete, and could be responsible for the particular stereochemical outcome of an experiment. PMID- 19300815 TI - New donor-acceptor chromophores by formal [2+2] cycloaddition of donor substituted alkynes to dicyanovinyl derivatives. AB - The efficient methodology of the cycloaddition between electron-rich alkynes and tetracyanoethylene (TCNE) or 7,7,8,8-tetracyanoquinodimethane (TCNQ), followed by retro-electrocyclisation, is extended to dicyanovinyl derivatives to produce new donor-acceptor push-pull 1,1-dicyanobutadienyl chromophores in excellent to quantitative yield (63-98%) that express strong charge-transfer (CT) absorptions from 300 to 600 nm. The scope of this reaction is established by both varying the nucleophilic and electrophilic components. Electrochemical studies show that the CT properties of these systems are readily tunable by substitution on the electrophile, which has the largest effect on the lowest unoccupied molecular orbital (LUMO). Non-reversible reduction potentials range from ca. -1.2 to -1.9 V in CH(2)Cl(2), against the ferricinium/ferrocene couple (Fc(+)/Fc) according to cyclovoltammetry (CV) and rotating disk voltammetry (RDV). The chromophores show a significant non-planarity between the N,N-dimethylanilino donor and the 1,1 dicyanovinyl acceptor moieties, with torsional angles around 40 degrees from X ray analysis, but retain strong quinoidal character. The mechanism of this reaction has been studied computational using density functional methods in the gas-phase and using the polarizable continuum model (PCM) for addressing solvent effects. The complete reaction free-energy profile has been determined for the reaction of 1,1-dicyanoethene and 4-ethynyl-N,N-dimethylaniline. The process proceeds through formal [2+2] cycloaddition followed by retro-electrocyclisation. The formation of a zwitterionic intermediate in the cycloaddition step is shown. PMID- 19300816 TI - Asymmetric synthesis of trans-disubstituted cyclopropanes using phosphine oxides and phosphine boranes. AB - The stereocontrolled synthesis of trans-disubstituted cyclopropylketones has been achieved from beta-alkyl, gamma-benzoyl phosphine oxides via a three-step cascade reaction incorporating an acyl transfer, phosphinoyl transfer and cyclisation to form the cyclopropane. Using Evans' chiral oxazolidinone auxiliary and by masking the phosphine oxide moiety as a phosphine borane we have extended the method to the synthesis of enantiomerically-enriched trans-disubstituted cyclopropyl ketones. PMID- 19300817 TI - The lithiation and acyl transfer reactions of phosphine oxides, sulfides and boranes in the synthesis of cyclopropanes. AB - Phosphine oxides are lithiated much faster than phosphine sulfides and phosphine boranes. Phosphine sulfides are in turn lithiated much more readily than phosphine boranes. It was possible to trap a phosphine sulfide THF in one case which upon treatment with t-BuOK gave cyclopropane, showing that phosphine sulfides readily undergo both phosphinoyl transfer and cyclopropane ring closure just like their phosphine oxide counterparts. The obtained data show that phosphine oxides are easily lithiated and undergo phosphoryl transfer much more readily and faster than phosphine sulfides and phosphine boranes. The observations suggest that it would be possible to perform reactions involving phosphine oxides in the presence of phosphine boranes or phosphine sulfides, potentially allowing regioselective alkylation of phosphine oxides in the presence of phosphine boranes or phosphine sulfides. PMID- 19300818 TI - [18F]- and [11C]-labeled N-benzyl-isatin sulfonamide analogues as PET tracers for apoptosis: synthesis, radiolabeling mechanism, and in vivo imaging study of apoptosis in Fas-treated mice using [11C]WC-98. AB - The radiolabeled isatin sulfonamide caspase-3 inhibitor, [18F] 2 (WC-II-89), is a potential PET radiotracer for noninvasive imaging of apoptosis. The radiolabeling mechanism was studied by 13C NMR, ESI/MS, and computational calculations. It was found that the high electrophilicity of the C3 carbonyl group in the isatin ring, which served as a trap for [18F]fluoride, was responsible for the failure of the radiolabeling via nucleophilic substitution of the mesylate group in 7a by [18F]fluoride. Once treated with a strong base, 7a opened the isatin ring completely to form an isatinate intermediate 16, which lost the ability to trap [18F]fluoride, thereby allowing the displacement of the mesylate group to afford the 18F-labeled isatinate 17. [18F] 17 can be converted to isatin [18F] 2 efficiently under acidic conditions. The ring-opening and re-closure of the isatin ring under basic and acidic conditions were confirmed by reversed phase HPLC analysis, ESI/MS and 13C NMR studies. Computational studies of model compounds also support the above proposed mechanism. Similarly, the ring-opening and re-closure method was used successfully in the synthesis of the 11C labeled isatin sulfonamide analogue [11C] 4 (WC-98). A microPET imaging study using [11C] 4 in the Fas liver apoptosis model demonstrated retained activity in the target organ (liver) of the treated mice. Increased caspase-3 activation in the liver was verified by the fluorometric caspase-3 enzyme assay. Therefore, this study provides a useful method for radio-synthesis of isatin derivative radiotracers for PET and SPECT studies, and [11C] 4 is a potential PET radiotracer for noninvasive imaging of apoptosis. PMID- 19300819 TI - Anthracene-DNA conjugates as building blocks of designed DNA structures constructed by photochemical reactions. AB - We present the photochemical ligation of anthracene-ODN (oligodeoxyribonucleotide) conjugates through cycloaddition of anthracenes. Anthracene-DNA conjugates were synthesized by linking the 5'- or 3'-end of ODNs to the anthracene group. The sequences of the conjugates were designed to hybridize to neighboring sites on the target ODN with their anthracene units facing each other. When the ternary duplex consisting of the two conjugates and the target forms, the conjugates can be dimerized by light irradiation. The dimerization efficiency was affected by the substitution position of the anthracene group and by a one-base displacement in the template sequence. Furthermore, it is demonstrated that the capping of the duplex and connecting third strands in a triplex structure could also be the target of the photochemical ligation. PMID- 19300820 TI - Convergent, stereoselective syntheses of the glycosidase inhibitors broussonetines D and M. AB - The first syntheses of the polyhydroxylated alkaloids (iminosugars) broussonetines D and M, glycosidase inhibitors of the pyrrolidine class, have been performed in a convergent, stereocontrolled way from D-serine as the chiral starting material. A cross metathesis step was one key feature of the synthesis. The versatility of the synthetic concept chosen permits the access to many members of this compound family, both natural ones and analogues thereof. PMID- 19300821 TI - Radical intermediates in chloroform reactions over triphenylphosphine-protected Au nanoparticles. AB - Reactions of chloroform over triphenylphosphine-protected Au nanoparticles have been studied using electron paramagnetic resonance (EPR) spectroscopy and a spin trapping technique. Two competing reactions, abstraction of hydrogen and halogen atoms, were identified. The hydrogen abstraction reaction showed an inverse kinetic isotope effect. Treatment of nanoparticles with oxidizing or reducing reagents made it possible to tune the selectivity of radical formation from halogen to hydrogen (deuterium) abstraction. Treatment with PbO(2) promoted the deuterium abstraction reaction followed by the loss of nanoparticle activity, whereas treatment with NaBH(4) regenerated the nanoparticle activity towards Cl atom abstraction. X-ray photoelectron spectroscopy showed an increased Au:P ratio upon treatment with oxidizing reagents. This is likely due to the oxidation of some phosphine ligands to phosphine oxides which then desorb from the nanoparticle surface. PMID- 19300822 TI - Investigation of acid-base catalysis in the extradiol and intradiol catechol dioxygenase reactions using a broad specificity mutant enzyme and model chemistry. AB - The extradiol and intradiol catechol dioxygenase reaction mechanisms proceed via a common proximal hydroperoxide intermediate, which is processed via different Criegee 1,2-rearrangements. An R215W mutant of extradiol dioxygenase MhpB, able to produce a mixture of extradiol and intradiol cleavage products, was analysed at pH 5.2-8.6, and the yield of extradiol product was found to be highly pH dependent, whereas the yield of intradiol product was pH-independent. The acid base chemistry of a biomimetic reaction for extradiol oxidative catechol cleavage was also investigated, using 1,4,7-triazacyclononane, FeCl(2), and pyridine in methanol, in which pyridine is proposed to act as both a general base and (in protonated form) a general acid. Kinetic experiments using a range of meta- and para-substituted pyridines gave a Bronsted plot of log(v) vs. pK(a) showing a bell-shaped plot. Oxidative catechol cleavage by a pyridine-monosubstituted beta cyclodextrin in the presence of TACN and FeCl(2) in methanol yielded only intradiol cleavage products. It is therefore proposed that bifunctional acid-base catalysis is required for iron (ii)-dependent extradiol catechol cleavage, whereas the rate-determining step for intradiol catechol cleavage does not involve acid-base catalysis. PMID- 19300823 TI - 8-Aza-7-deazaguanine nucleosides and oligonucleotides with octadiynyl side chains: synthesis, functionalization by the azide-alkyne 'click' reaction and nucleobase specific fluorescence quenching of coumarin dye conjugates. AB - Oligonucleotides incorporating 7-(octa-1,7-diynyl) derivatives of 8-aza-7-deaza-2 deoxyguanosine (2d) were prepared by solid-phase synthesis. The side chain of 2d was introduced by the Sonogashira cross coupling reaction and phosphoramidites (3a, 3b) were synthesized. Duplexes containing 2d are more stabilized compared to those incorporating the non-functionalized 8-aza-7-deaza-2-deoxyguanosine (2a) demonstrating that these side chains have steric freedom in duplex DNA. Nucleoside 2d as well as 2d-containing oligonucleotides were conjugated to the non-fluorescent 3-azido-7-hydroxycoumarin 15 by the Huisgen-Meldal-Sharpless click reaction. Pyrazolo[3,4-d]pyrimidine nucleoside conjugate 16 shows a much higher fluorescence intensity than that of the corresponding pyrrolo[2,3 d]pyrimidine derivative 17. The quenching in the dye conjugate 17 was found to be stronger on the stage of monomeric conjugates than in single-stranded or duplex DNA. Nucleobase-dye contact complexes are suggested which are more favourable in the monomeric state than in the DNA chain when the nucleobase is part of the stack. The side chains with the bulky dye conjugates are well accommodated in DNA duplexes thereby forming hybrids which are slightly more stable than canonical DNA. PMID- 19300824 TI - Synthesis of azepino[4,5-b]indolones via an intermolecular radical oxidative substitution of N-Boc tryptamine. AB - A two-step protocol for the synthesis of azepino[4,5-b]indolone derivatives featuring a xanthate radical oxidative aromatic substitution on the N-Boc protected tryptamine, using dilauroyl peroxide (DLP) as initiator and oxidant, is described. PMID- 19300825 TI - Isolation of the key intermediates in the catalyst-free conversion of oxiranes to thiiranes in water at ambient temperature. AB - Herein we shed light on the mechanism of the reaction of epoxides with ammonium thiocyanate to give the corresponding thiiranes in water, and we present a computational mechanistic model for this highly useful reaction. PMID- 19300826 TI - Efficient enzymatic synthesis of LNA-modified DNA duplexes using KOD DNA polymerase. AB - Three different LNA-nucleoside triphosphates, LNA-TTP, LNA-ATP and LNA-5-methyl CTP, were investigated as substrates for KOD DNA polymerase. The results reveal that KOD DNA polymerase is an efficient catalyst for template directed synthesis of DNA oligonucleotide duplexes containing a large number of LNA nucleotides by primer extension reactions. Furthermore, KOD DNA polymerase is shown to be suitable for the PCR amplification of an LNA-modified DNA duplex. PMID- 19300827 TI - A new mild base-catalyzed Mannich reaction of hetero-arylamines in water: highly efficient stereoselective synthesis of beta-aminoketones under microwave heating. AB - A new mild base-catalyzed Mannich reaction of aromatic aldehydes with 1,2 diphenylethanone and hetero-arylamines including pyridin-2-amine and pyrimidin-2 amine is described. In this reaction, a series of new beta-aminoketones were stereoselectively synthesized in water by controlling the steric hindrance of the substrates under microwave heating. This method has the advantages of a short synthetic route, operational simplicity, increased safety for small-scale high speed synthesis, and minimal environmental impact. PMID- 19300828 TI - Synthesis and anti-HIV activity of conformationally restricted bicyclic hexahydroisobenzofuran nucleoside analogs. AB - A chiral synthesis of a series of hexahydroisobenzofuran (HIBF) nucleosides has been accomplished via glycosylation of a stereo-defined (syn-isomer) sugar motif 16 with the appropriate silylated bases. All nucleoside analogs were obtained in 52-71% yield as a mixture of alpha- and beta-anomeric products increasing the breadth of the novel nucleosides available for screening. The structure of the novel bicyclic HIBF nucleosides was established by a single crystal X-ray structure of the beta-HIBF thymine analog 22b. Furthermore, the sugar conformation for these nucleosides was established as N-type. Among the novel HIBF nucleosides synthesized, twenty-five compounds were tested as inhibitor of HIV-1 in human peripheral blood mononuclear (PBM) cells and seven were found to be active (EC(50) = 12.3-36.2 microM). Six of these compounds were purine analogs with beta-HIBF inosine analog 22o being the most potent (EC(50) = 12.3 microM) among all compounds tested. The striking resemblance between didanosine (ddI) and 22o may explain the potent anti-HIV activity. PMID- 19300829 TI - Synthesis of spiroacetal-nucleosides as privileged natural product-like scaffolds. AB - The elaboration of a 6,6-spiroacetal scaffold to incorporate a nucleoside unit at the anomeric position is described. The novel spiroacetal-nucleoside hybrids were generated via nucleosidation of acetoxy-spiroacetal with a series of silylated nucleobases under Vorbruggen conditions. PMID- 19300830 TI - Molecular recognition in anisotropic media. Binding of alkylpyridines to amphiphilic zinc porphyrins incorporated in liposomal bilayer membranes. AB - Two amphiphilic zinc porphyrins were incorporated into liposomal bilayer membranes, egg phosphatidylcholine (Egg PC) and dipalmitoylphosphatidylcholine (DPPC). Binding free energy of alkylpyridines to the zinc porphyrins linearly increased as the length of the alkyl chains of the guest increased, showing that the guest was incorporated deep in the bilayer membranes in a hydrophobic environment. Comparison of the free energy increase per CH(2) group indicated that recognition of 3-alkylpyridines were favored over that of 4-alkylpyridines by deltaDeltaDeltaG(o) = 0.3-0.7 kJ mol(-1), and this preference was attributed to the anisotropy of the liposomal bilayer membranes. The binding was exothermic (DeltaH(o) = -15 to -21 kJ mol(-1)) when the liposome was in a liquid crystalline phase, while it was endothermic (DeltaH(o) = 53 to 62 kJ mol(-1)) when the liposome was in a gel phase. Local disorder of lipid molecules may be a driving force for binding in the latter case. Lipid bilayer membranes provide a unique medium for molecular recognition, in which the anisotropy and entropy of the lipid molecules add new features to binding. PMID- 19300831 TI - Redox active donor-substituted punicin derivatives. AB - The redox active plant material punicin from Punica granatum, 2-hydroxy-1 (pyridinium-1-yl)-5-olate, and some derivatives were modified by substitution with 2-oxochromen-4-olate moieties to give donor-substituted molecules which form distinct types of atropisomeric mesomeric betaines and tetrapolar substances. Oxidation to new negatively-charged solvatochromic quinones was achieved on treatment of these betaines with CAN, taking advantage of the stabilizing properties of the electron-donating 2-oxochromen-4-olate partial structure. The quinones can be employed in model redox reactions. Cyclovoltametric studies have been performed. PMID- 19300832 TI - New glycosylated derivatives of versipelostatin, the GRP78/Bip molecular chaperone down-regulator, from Streptomyces versipellis 4083-SVS6. AB - Four novel glycosylated derivatives of versipelostatin (1), versipelostatins B-E (2-5), were isolated from the culture broth of Streptomyces versipellis 4083 SVS6. The inhibitory activities of the isolated compounds against the expression of molecular chaperone GRP78 induced by 2-deoxyglucose were evaluated. Of the five versipelostatin family members, 1 and 4 were the more potent with IC(50) values of 3.5 and 4.3 microM. These results suggest that the alpha-L oleandropyranosyl (1-->4)-beta-D-digitoxopyranosyl residue in the sugar moiety may play an important role in down-regulating GRP78 expression induced by 2 deoxyglucose. PMID- 19300833 TI - Exploring the use of APTS as a fluorescent reporter dye for continuous glucose sensing. AB - The anionic fluorescent dye, aminopyrene trisulfonic acid (APTS), was synthesized and used in a solution-based two-component glucose-sensing system comprising the dye and a boronic acid-appended viologen. The fluorescence of the dye was quenched in the presence of the viologen and the fluorescence restored upon glucose addition. An important feature of this fluorophore is that it can be covalently bonded to a polymer through the amine group without a significant effect on optical properties. Two APTS derivatives, functionalized with polymerizable groups, were synthesized and immobilized in hydroxyethyl methacrylate (HEMA)-based hydrogels. The latter were used to continuously monitor glucose. The fluorescence signal modulation, signal stability, reversibility, reproducibility, and pH sensitivity of the hydrogels were evaluated. The APTS dyes described herein are insensitive to pH changes within the physiological range, both in solution and when immobilized in a hydrogel. When APTS is used in conjunction with boronic acid-appended viologens to sense glucose, the system displays some pH sensitivity because of the presence of the boronic acid. PMID- 19300834 TI - On the origin of the regioselectivity in glycosylation reactions of 1,2-diols. AB - The assistance of neighboring protecting groups with different orientations in 1,2-diol acceptors and the reactivity of both reaction partners, the donor and the acceptor, have been evaluated as factors that determine the regioselectivity of glycosylation reactions. It has been established, by experimental and theoretical studies, that the regioselectivity for the glycosylation of a given OH group can be considerably increased by the presence of groups able to form a hydrogen bond with that OH group. Moreover higher regioselectivities are observed when armed donor/activated acceptor combinations are avoided. PMID- 19300835 TI - [Media, morality, and risk factors in health]. PMID- 19300836 TI - [Human functioning and disability: exploring the scope of the World Health Organization's international classification]. AB - The theoretical discussion on disability is dichotomized according to the medical and social perspectives. The biomedical model focuses on impairment, disease, or physical abnormality and how these factors produce disability. The social approach suggests that the meaning of disability and impairment emerges from specific social and cultural contexts. The WHO created the International Classification of Functioning, Disability and Health (ICF), with a classification system and theoretical model based on the combination of the medical and social models and using a biopsychosocial approach to integrate the health dimensions. Despite the importance and immediacy of the ICF, some concepts were insufficiently detailed and justified and could lead to distinct interpretations. This essay proposes to describe the ICF model and analyze the scope of the biopsychosocial theory for exploring the relational nature of the 'disability' and 'impairment' categories, as well as the universal nature of the WHO proposal. One of the most positive aspects of the ICF is to highlight the interactive nature of disability and the division of the phenomenon into three dimensions, thus demonstrating the degree of complexity in the process of human functioning and disability. PMID- 19300837 TI - [Public policy to control violence: a case study in Salvador, Bahia State, Brazil]. AB - The aim of this study was to analyze the formulation and preliminary developments of a public policy to control violence in Salvador, Bahia State, Brazil. The study's theoretical reference was the cycle of public policies and contributions by 'rational choice'. The public policy analyzed here reached the first three levels of the political decision-making process: the issue was placed on the public agenda and was present in the discourse of public authorities, and a plan was drafted at the municipal level. The fact that various stakeholders and sectors were convened in the organizational structure proposed by the plan was not sufficient to encourage cooperation among individuals to act jointly in the plan's implementation. The mechanisms used to sensitize the stakeholders were not sufficient to mobilize them for the fight against violence under the proposed format. A public policymaking process should also consider the set of factors that encourage or constrain cooperation among individuals for collective action from the perspective of the common good. PMID- 19300838 TI - [Pharmacy records as an indicator of non-adherence to antiretroviral therapy by HIV-infected patients]. AB - The objectives of this study were to evaluate anti-retroviral (ARV) prescription pickups during twelve months following the first prescription and to identify factors associated with irregular pickups or permanent dropout in two public HIV/AIDS referral centers in Belo Horizonte, Minas Gerais State, Brazil. Participants (n = 323) were antiretroviral naive and were recruited from May 2001 to May 2002. A total of 98 (30.3%) patients abandoned treatment, and 187 (57.9%) had at least one irregular pickup. Patients with irregular pickups and dropouts were compared to those with regular pickups. Multinomial multivariate analysis showed that living outside Belo Horizonte, CD4+ T-lymphocyte count greater than 200/mm(3), and antiretroviral regimen without protease inhibitors were associated with irregular pickups. In addition to these variables, not being on other medications, and any non-adherence recorded on patient charts were associated with treatment dropout. Pharmacy records are important potential indicators of non-adherence and should be incorporated as such in clinical practice. Strategies should be prioritized to reach out to dropouts or patients with irregular ARV pickups. PMID- 19300839 TI - [Screening for risk of hospitalization in the elderly: a study based on a single entry point in a health maintenance organization in Brazil]. AB - Population aging requires new strategies for the evaluation and care of different elderly groups. The aim of this study was to present a single entry point model for health care to the elderly in a health maintenance organization. The model prioritizes care for individuals at greatest risk, defining the groups followed subsequently under a specific program for prevention and care. Telephone interviews were used to evaluate the probability of hospitalization for 2,637 users of the health plan. 53.9% of the individuals who were contacted agreed to participate in the survey, and an index was calculated to estimate the probability of hospitalization. 3.23% of subjects were classified as high risk, 7.23% as medium-high risk, and 13.4% as medium risk. Subsequent prioritization of care was based on this stratification. The model will enable improved planning and definition of priorities for resource allocation within a health care system subject to serious funding limitations. Studies that identify criteria for the distribution of health care resources are thus essential to the success of consistent and realistic health policies for the elderly. PMID- 19300840 TI - [Public knowledge on the role of physical activity in the prevention and treatment of diabetes and hypertension: a population-based study in southern Brazil]. AB - The aim of this study was to evaluate public knowledge on the role of physical activity in the prevention and treatment of diabetes and hypertension, and the factors associated with such knowledge. A population-based cross-sectional study was conducted in Pelotas, southern Brazil, including 972 adults aged 20 to 69 years, selected with a clustering protocol. Knowledge on the preventive and curative benefits of physical activity was higher for hypertension (87.2%) than for diabetes (47.2%). Women were more knowledgeable on the role of physical activity in preventing diabetes (PR: 1.16; 95%CI: 1.03-1.31). In terms of treatment, greater knowledge was associated with female gender, current physical activity, obesity, subjects, and higher socioeconomic status. For prevention of hypertension, greater knowledge was observed in individuals with higher socioeconomic status (PR: 1.23; 95%CI: 1.11-1.36). For treatment of hypertension, physically active and obese subjects showed greater knowledge. Subjects were generally more knowledgeable on the curative role of physical activity than on its preventive benefits. Public health efforts should aim to raise public awareness on the preventive effects of physical activity against diabetes, hypertension, and other chronic non-communicable diseases. PMID- 19300841 TI - [Health-related behavior in a sample of Brazilian college students: gender differences]. AB - This study investigated whether undergraduate students' health-risk behaviors differed according to gender. The sample consisted of 382 subjects, aged 20-29 years, from public universities in Pernambuco State, Brazil. Data were collected using the National College Health Risk Behavior Survey, previously validated in Portuguese. Descriptive and inferential statistical techniques were used. Associations were analyzed with the chi-square test or Fisher's exact test. Statistical significance was set at p < or = 0.05. In general, females engaged in the following risk behaviors less frequently than males: alcohol consumption (p = 0.005), smoking (p = 0.002), experimenting with marijuana (p = 0.002), consumption of inhalants (p < or = 0.001), steroid use (p = 0.003), carrying weapons (p = 0.001), and involvement in physical fights (p = 0.014). Meanwhile, female students displayed more concern about losing or maintaining weight, although they exercised less frequently than males. The findings thus showed statistically different health behaviors between genders. In conclusion, different approaches need to be used for the two genders. PMID- 19300842 TI - [Effect of exposure time to PM(10) on emergency admissions for acute bronchitis]. AB - To study the health effect of air pollution, measured as particulate matter greater than 10mm in diameter (PM(10)), we analyzed the effect of daily hours of exposure on the number of urgency admissions for acute bronchitis, adjusting for temperature and humidity on the same day. The study was conducted in six regions of Santiago, Chile, during the winter of years 2002 to 2004, for infants and elders. The delay between pollution time series and disease was modeled using a polynomial distributed lag (PDL) function included in a generalized linear model. The linearity assumption was evaluated using a smooth-spline model approach. The highest effect for exposure to PM(10) was detected with 4 days of delay. For both groups, the effect of temperature was linear, but that of humidity was not. Air pollution effect varied according to level of exposure and geographic region, increasing by 3% the total number of infant emergency consultations for each hour of exposure to levels greater than 150 microg/m(3). The study thus showed a significant impact of air pollution on population health. PMID- 19300843 TI - [Characteristics of the medical workforce in the Province of Cabinda, Angola]. AB - This article presents the results of a study on the public health system's medical workforce in the Province of Cabinda, Angola. The objective was to study the characteristics of medical personnel, seeking to help define strategies to expand access and improve quality of care. The study covered the period from 2001 to 2004 and used secondary data, semi-structured interviews with health professionals, direct observation, and comparison with the literature. Quality of medical care in Cabinda was low, with one physician per 3,356 inhabitants. There were communities with more than 35,000 inhabitants and not a single physician. Working conditions were poor, and physicians suffered from low motivation. The study concluded that formulation and implementation of policies envisioning improvements in the management of medical professionals in the sector could help enhance the quality of care in the Province, but it would require involvement by relevant actors and additional research on the other health professions. PMID- 19300844 TI - [Sexual behavior and emergency contraception among adolescents from public schools in Pernambuco State, Brazil]. AB - This study focused on knowledge and use of emergency contraception among 4,210 adolescents (14-19 years) enrolled in public schools in Pernambuco State, Brazil. Information was collected using the Global School-Based Student Health Survey, previously validated. Knowledge, frequency, and form of use of emergency contraception were investigated. Independent variables were classified as socio demographic and those related to sexual behavior. Most adolescents reported knowing and having received information about the method, but among those who had already used it, only 22.1% had done so correctly. Adjusted regression analysis showed greater likelihood of knowledge about the method among girls (OR = 5.03; 95%CI: 1.72-14.69) and the sexually initiated (OR = 1.52; 95%CI: 1.34-1.75), while rural residents were 68% less knowledgeable. Rural residents showed 1.68 times higher odds (CI95%: 1.09-2.25) of incorrect use, while girls showed 71% lower likelihood of incorrect use. Sexual and reproductive education is necessary, especially among male and rural adolescents. PMID- 19300845 TI - Evaluation of Bacillus sphaericus bioinsecticide produced with white soybean meal as culture medium for the control of Culex (Culex) quinquefasciatus. AB - Bioinsecticides are shown to be useful in control programs to prevent several diseases, based on their specificity and efficiency against insect vectors. In the current study a bioinsecticide based on Bacillus sphaericus was produced using a white soybean culture medium and applied to larvae of Culex quinquefasciatus, the susceptible species, and Aedes aegypti, the refractory species used as the negative control. Efficacy was compared with that of the product fermented with the Luria Bertani (LB) reference medium. The experiments showed that C. quinquefasciatus was highly susceptible to the product prepared with white soybean meal, reaching 100% larval mortality even at 10mg/L, while A. aegypti failed to reach 70% mortality at a concentration of 1g/L. By comparison with the reference medium, the proposed culture medium showed high larvicidal power, reaching a LD90 of 2.26 mg/L, while 4.37 mg/L was needed for the LB medium to achieve the same mortality rate. Cost comparison between the formulations favored the use of the bioinsecticide produced with white soybean meal. After factoring in the LD90 value, the cost ratio favored the new raw material by nearly 1:220. PMID- 19300846 TI - [Factors associated with overweight and abdominal fat in adults in Salvador, Bahia State, Brazil]. AB - This cross-sectional study of 570 adults aimed to identify factors associated with overweight and abdominal fat in adults of both sexes in the city of Salvador, Bahia State, Brazil. Body mass index (BMI) and waist circumference were the dependent variables, and Poisson regression was used for statistical analysis. For women, high BMI and waist circumference were associated with increased age and current dieting (< or = 3 meals a day), while family history of obesity was only associated with increased BMI. In men, BMI increased in the 30 39 and 50-59-year age brackets. Waist circumference increased in association with < or = 3 meals a day, arterial hypertension, and alcohol consumption, and was lower among blacks and mixed-race individuals. High BMI and waist circumference were both associated with family history of myocardial infarction. Thus, factors associated with overweight and abdominal fat differ between men and women, but adopting healthier lifestyles can help modify many of these factors. PMID- 19300847 TI - [Quality of data on the Child Health Record and related factors]. AB - The Child Health Record (CHR) is an essential document for children's health care activities. A cross-sectional study was performed with simple random sampling, aimed at analyzing factors related to quality of data completion on the CHR. Interviews and direct verification of 365 CHRs from 9 health districts in Belo Horizonte, Minas Gerais State, Brazil, were performed. A scoring system was created to classify the CHRs in terms of quality of data completion and to relate them to explanatory variables. Odds ratios were calculated by logistic regression. Completion of the 20 scoring items varied from 3.1% (use of iron supplements) to 99.7% (date of birth). Factors associated with worse scores were: children > 12 months old (OR = 1.77), mothers with < or = 6 years of schooling (OR = 1.97), children not treated by general practitioners (OR = 3.18), and mothers who had not received explanations on the CHR while in the maternity ward (OR = 1.77). The results show poor use of the CHR and emphasize the need for on going efforts to train health professionals and prepare health services for their important role in the promotion of children's health. PMID- 19300848 TI - [Factors associated with duration of breastfeeding for children of low-income families from southern Curitiba, Parana State, Brazil]. AB - Various studies have focused on breastfeeding and the possible factors associated with early weaning. This study aimed to identify risk factors and protective factors for duration of breastfeeding in low-income families in southern Curitiba, Parana State, Brazil. A cohort study from November 2006 to March 2007 included 118 mothers of children less than two years of age and enrolled in the TC Health Unit (USTC) located in the southern area of Curitiba. Statistical analyses using the Kaplan-Meier method and Cox regression model identified the following protective factors for breastfeeding: mother's awareness of the benefits of extended breastfeeding, exclusive feeding with mother's breast milk while in the maternity ward, and rooming-in. Factors contributing to early weaning were low birth weight, mother's work outside the home, and difficulties experienced by the mother in breastfeeding during the first days postpartum. PMID- 19300849 TI - A population based study on health conditions associated with the use of benzodiazepines among older adults (The Bambui Health and Aging Study). AB - The aim of this study was to assess the health conditions and use of health services among elderly users of benzodiazepines. The study was carried out in the city of Bambui in Minas Gerais State, Brazil. Of 1,742 inhabitants aged over 60, 1,419 participated in the study. Information about benzodiazepine and the use of other medications was obtained by means of interviews and checking medication packaging. After adjustments for confounding variables, benzodiazepine remained significantly associated with poorer self reported health (prevalence ratio = 1.94 and 2.04 for reasonable and bad/very bad health, respectively), common mental disorders (1.33), myocardial infarction (1.42), number of doctor visits in the previous 12 months (1.90 and 2.24 for 2-3 and 4+ visits, respectively) and concomitant use of other medications (1.84 and 1.83 for 2-4 and 5+ medications). The results showed that the factors associated with the use of benzodiazepine were similar to those observed in the elderly in higher income countries. The concomitant use of benzodiazepines and two or more medications was highly prevalent (59.5%), characterizing a situation of risk, which deserves attention as a public health problem. PMID- 19300850 TI - [Evaluation of the completeness of variables on Birth Certificates of residents in Pernambuco State, Brazil, 1996 to 2005]. AB - An evaluative, descriptive, time series study was conducted to evaluate the completeness of data on certificates of live birth. The study population consisted of all live births of residents in Pernambuco State, Brazil, from 1996 to 2005, contained in the database of the Live Birth Information System (SINASC), provided by the Information Technology Department of the Unified National Health System (DATASUS). We calculated each variable's incompleteness (blank/unknown data) by municipality of residence. Based on these data, we calculated the percentage of municipalities with excellent completeness (> 95%), constructed a simple linear regression model, and calculated the determination coefficient. An important increase was observed in the percentage of municipalities with excellent completeness, highlighting the following variables: race/color, maternal schooling, maternal marital status, and number of prenatal visits. The study emphasizes the importance and relevance of using the SINASC database for analyzing maternal and child health conditions. PMID- 19300851 TI - [Provision of voluntary surgical sterilization in the Campinas Metropolitan Area, Sao Paulo State, Brazil: perceptions of public health services managers and professionals]. AB - This study describes the perceptions of public health services managers and professionals concerning provision of voluntary surgical sterilization in the Campinas Metropolitan Area, Sao Paulo State, Brazil. The study adopted a qualitative approach in four municipalities (counties), where semi-structured interviews were conducted with 26 health professionals and health services managers involved in the provision of surgical sterilization. The interviewees identified difficulties in scheduling visits at Outpatient Family Clinics or Reference Centers (APF/CR), and the number of available surgeries in the accredited hospitals was insufficient. They emphasized the lack of physical infrastructure and human resources for conducting family planning activities in the primary health units as well as in the APF/CR.They also criticized the legal criteria for authorizing surgical sterilization, and mentioned adaptations to make them more appropriate to the each municipality's situation. According to the health services managers and professionals, despite the efforts, meeting the demand for surgical sterilization in the Campinas Metropolitan Area was jeopardized by its centralization in the APF/CR, which in practice had to cover the gap in family planning activities in each municipality's primary care units. PMID- 19300852 TI - Air quality and emergency pediatric care for symptoms of bronchial obstruction categorized by age bracket in Rio de Janeiro, Brazil. AB - The objective of this study was to estimate the association between levels of air pollutants and respiratory symptoms in children. An ecological time-series study was conducted between April 2002 and March 2003 with daily data on PM10, SO2, CO, NO2, and O3 and paediatric care in emergency rooms due to symptoms indicating bronchial obstruction. The potential confounding factors considered were: time trend, seasonality, meteorological variables, respiratory infections and the effects of weekends and holidays. A semi-parametric Poisson regression was used to model the time series. Splines (data smoothing functions), indicator variables and cubic polynomials were used to adjust the effects of the confounding variables. A 5% significance level was adopted for the study. A statistically significant increase of 6.7% in paediatric visits of children of less than 2 years of age was associated with PM10. A 3% positive association with O3 showed borderline significance (p < 0.06) in this age bracket. Our findings highlight the existence of an acknowledged public health problem in Rio de Janeiro, Brazil, and also emphasize the need to identify the principal sources of air pollutants. PMID- 19300853 TI - [Objective and perceived knowledge of oral contraceptive methods among adolescent mothers]. AB - The high rate of early pregnancy in Brazil and particularly in Teresina (21.5%), Piaui State, motivated the current study, the aim of which was to identify levels of objective and perceived knowledge on oral contraceptives, as well as predictive reproductive and socio-demographic variables for high knowledge. A cross-sectional study was performed including 278 teenage mothers (15-19 years), with their childbearing history, admitted to four maternity hospitals in Teresina in 2006. Logistic regression provided the basis for the statistical analysis. Nearly 98% of the adolescent mothers showed low objective and perceived knowledge of oral contraceptives. High parity was the only predictor of increased objective knowledge on oral contraceptives. The adolescents' low level of objective and perceived knowledge on use of oral contraceptives emphasized their susceptibility to risky sexual behavior. The findings emphasize the need for a more interactive approach with adolescents concerning their level of objective and perceived knowledge, to reduce the occurrence and repetition of unwanted teenage pregnancy and its negative consequences for the lives of these young mothers and their children. PMID- 19300854 TI - [Factors associated with oral health habits and use of dental services by adolescents]. AB - This was a cross-sectional study of 1,170 seventh-grade adolescents from the municipal public school system in Gravatai, Rio Grande do Sul State, Brazil, investigating the association between socio-demographic, psychosocial, and lifestyle factors and oral health habits and use of dental services. Data were analyzed by Cox regression, modified for cross-sectional studies. Females showed higher frequency of brushing, as did adolescents that reported not feeling alone or discriminated. Daily flossing was associated with higher socioeconomic status (SES), use of private dental services, parental understanding, and lack of feeling of loneliness. Frequency of annual dental visits was higher among individuals with higher SES. Preventive dental checkups were more frequent among individuals with higher SES, those who felt understood by their parents, and those who did not habitually eat candy. Oral health habits were associated with family SES and psychosocial factors except for frequency of annual dental visits. As for lifestyle, low candy consumption had a positive impact on reasons for use of dental appointments. PMID- 19300855 TI - Prevalence and risk factors for HIV, syphilis, hepatitis B, hepatitis C, and HTLV I/II infection in low-income postpartum and pregnant women in Greater Metropolitan Vitoria, Espirito Santo State, Brazil. AB - Sexually transmitted infections during pregnancy pose a major risk to the fetus due to vertical transmission. The study's objective was to determine the prevalence of HIV, syphilis, hepatitis B and C, and HTLV-I/II infection among low income postpartum and pregnant women treated in Greater Metropolitan Vitoria, Espirito Santo State, Brazil, and the risk factors associated with these infections. A cross-sectional study was conducted from February to October 1999 assessing postpartum and pregnant women from the maternity ward of the Vitoria Mercy Hospital and the Carapina Outpatient Referral Unit in the Municipality of Serra, respectively. Patients were systematically interviewed and had blood samples drawn for serological tests (HIV 1&2, VDRL, HbsAg, anti-HCV, and HTLV I/II). A total of 534 patients (332 postpartum and 202 pregnant women) were assessed. Seroprevalence rates for the target infections in postpartum and pregnant women and the overall sample were as follows, respectively: HIV 0.9%, 0%, and 0.6%; syphilis 2.1%, 3.6%, and 2.7%; HBV 1.2%, 1%, and 1.1%; HCV 1.8%, 0.6%, and 1.4%; and HTLV-I/II 1.7%, 0.6%, and 1.3%. Factors associated with the various infections are presented and analyzed in light of other research findings from the literature. PMID- 19300856 TI - Forum: challenges in STD/AIDS prevention in Portuguese-speaking African countries: contributions from social research and from a gender approach. Introduction. AB - This forum on the challenges of preventing STD/AIDS in Portuguese-speaking African countries contains three articles and a postscript. The first paper reviews academic production on the topic from the fields of the social sciences and of health, with special attention on how local cultural and socioeconomic factors impact the dynamics of the epidemic. Based on an ethnographic study of a region in southern Mozambique, the second paper analyzes the notion of 'tradition' within the context of Mozambique and how it affects perceptions of the local population's vulnerability to STD/AIDS. The third and final article discusses common ground and differences between government and civil society in gender approaches by community HIV/AIDS projects in Mozambique. Their observations suggest that important mistakes have been made in STD/AIDS prevention discourse and initiatives in African countries because the unique features of local development models and cultural systems have not been taken into account. PMID- 19300857 TI - STD/AIDS prevention in Portuguese-speaking Africa: a review of the recent literature in the social sciences and health. AB - The article reviews academic literature in the social sciences and health on the problems and challenges of STD/AIDS prevention in Portuguese-speaking African countries. Based on a bibliographic survey of the SciELO, PubMed, and Sociological Abstracts databases between 1997 and 2007, the research under review was organized into two groups, according to content. The first group of studies sought to understand STD/AIDS vulnerability among social groups by examining local cultural and socioeconomic factors as related to gender dynamics, sexuality, color/race, religion and health care. The second group encompassed critical assessments of shortcomings in the STD/AIDS educational messages delivered by governments and international agencies. Attention is called to the way in which the presence of traditional medicine systems and the occurrence of civil wars in the post-colonial period impact the STD/AIDS epidemic in the African countries under study. PMID- 19300858 TI - 'Tradition', person, gender, and STD/HIV/AIDS in southern Mozambique. AB - In southern Mozambique, the "traditional" notion of personhood is constructed through a process, as an outcome of diachronic and synchronic social relations that encompass kin and other peers, including spirits. Both person and body are thought of as elements traversed and determined by these relations, which include the gender relations whose complementarity finds expression in alliances and the production of descendants. In this system of agnatic kinship, descent is possible through women, who produce the male and female persons. Because of women's structural position, they may be suspected of fostering deconstruction of the person as well, with diseases providing the objective data that ground such a charge. To a certain degree, HIV/AIDS has been experienced in terms of this sociocultural arrangement, which defines disease as the result of action by social subjects that jeopardizes the person, placing women in the vulnerable position of being seen as the producers of disease. This has defined the ways in which people experience both the epidemic as well as STD/HIV/AIDS prevention and treatment messages and public policies. PMID- 19300859 TI - The gender approach in community AIDS projects in Mozambique: agreement and disagreement between government and civil society. AB - This article discusses some areas where government and civil society converge and clash in their gender approaches in community HIV/AIDS projects in Mozambique, based on an evaluative study conducted in 2006 encompassing 160 of the 1,124 NGO projects undertaken with the support of the country's national AIDS council, known as the Conselho Nacional de Combate ao SIDA (CNCS). An analysis of projects and official documents shows that, for the CNCS, the term 'gender' represents a way of underscoring the epidemic's impact on women. In community projects, the gender approach often times finds expression in initiatives to mitigate the economic impact of the epidemic on widows. Initiatives aimed at men and at the population as a whole generally pay little attention to power relations between men and women or their affect on the epidemic. This suggests that any endeavor to transfer Western analytical techniques or forms of intervention for coping with the HIV/AIDS epidemic to other regions of the world demands painstaking efforts to translate these and adapt them to local cultural standards. PMID- 19300860 TI - Forum: challenges in STD/AIDS prevention in Portuguese-speaking African countries: contributions from social research and from a gender approach. Postscript. PMID- 19300861 TI - Oral health in the family health strategy: a change of practices or semantics diversionism. AB - OBJECTIVE: To evaluate public health dentistry practices of two different family health models. METHODS: Qualitative study conducted with data obtained from focus groups consisting of 58 dentists working in the Family Health Strategy for at least three years between August-October, 2006. The Paideia Family Health Approach was used in the city of Campinas and the Oral Health Initiative as part of the Family Health Strategy was implemented in the city of Curitiba, Southeastern and Southern Brazil, respectively. Data was analyzed using the hermeneutic-dialectic method. Analysis indicators were employed to indicate backwardness, stagnation or progress in oral health practices effective from the implementation of the strategies referred. The indicators used were: work process; interdisciplinary approach; territorialization; capacity building of human resources; health promotion practices; and responsiveness to users' demands. RESULTS: There was progress in user access to services, humanization of health care, patient welcoming and patient-provider relationship. The results related to health promotion practices, territorialization, interdisciplinary approach and resource capacity building indicated a need for technical and operational enhancements in both cities. CONCLUSIONS: Both models have brought about important advances in terms of increased access to services and humanization of health care. Universal access to oral health at all levels of complexity was not achieved in both cities studied. Local health managers and oral health program coordinators must bring more weight to bear in the arena that defines public policy priorities. PMID- 19300863 TI - Metabolic response to a ketogenic breakfast in the healthy elderly. AB - OBJECTIVE: To determine whether the metabolism of glucose or ketones differs in the healthy elderly compared to young or middle-aged adults during mild, short term ketosis induced by a ketogenic breakfast. DESIGN AND PARTICIPANTS: Healthy subjects in three age groups (23 +/- 1, 50 +/- 1 and 76 +/- 2 y old) were given a ketogenic meal and plasma beta -hydroxybutyrate, glucose, insulin, triacylglycerols, total cholesterol, non-esterified fatty acids and breath acetone were measured over the subsequent 6 h. Each subject completed the protocol twice in order to determine the oxidation of a tracer dose of both carbon-13 (13C) glucose and 13C-beta-hydroxybutyrate. The tracers were given separately in random order. Apolipoprotein E genotype was also determined in all subjects. RESULTS: Plasma glucose decreased and beta-hydroxybutyrate, acetone and insulin increased similarly over 6 h in all three groups after the ketogenic meal. There was no significant change in cholesterol, triacylglycerols or non esterified fatty acids over the 6 h. 13C-glucose and 13C-beta-hydroxybutyrate oxidation peaked at 2-3 h postdose for all age groups. Cumulative 13C-glucose oxidation over 24 h was significantly higher in the elderly but only versus the middle-aged group. There was no difference in cumulative 13C-beta-hydroxybutyrate oxidation between the three groups. Apolipoprotein E (epsilon 4) was associated with elevated fasting cholesterol but was unrelated to the other plasma metabolites. CONCLUSION: Elderly people in relatively good health have a similar capacity to produce ketones and to oxidize 13C-beta-hydroxybutyrate as middle aged or young adults, but oxidize 13C-glucose a little more rapidly than healthy middle-aged adults. PMID- 19300864 TI - Impact of food aid on food variety and dietary diversity of an elderly community in Sharpeville, South Africa. AB - OBJECTIVE: The objective of this paper was to discuss the impact of a two-year food aid intervention programme on dietary diversity and adequacy of an elderly community. This project was undertaken amongst the elderly (n=300), voluntarily attending a day care centre in Sharpeville, South Africa. In a baseline study (2004) in this community the dietary diversity scoring (DDS) method revealed low dietary diversity (3.41) and food variety (4.77) scores (FVS), and a trend that higher scores resulted in a better mean nutrient adequacy ratio. An intervention study, implemented in 2005, aimed to improve the dietary diversity of this elderly community by providing breakfast and lunch five days a week at the care centre. The impact of this food aid was investigated by comparing the DDS, FVS and nutrient adequacy ratios (NAR) before and after the intervention. METHODS: The methods included socio-demographic, health, food frequency questionnaire (FFQ), 24 hour-recall (24h-recall), anthropometric and biochemical measurements in 107 elderly people who regularly attended the care centre since 2004. RESULTS: At baseline, the majority of respondents (n=82, 55.1%) were classified with low DDS utilizing 0-3 food groups compared to after the intervention, where 98.1% (n=105) was classified with a high dietary diversity utilizing all nine nutritious food groups. The mean individual DDS of 3.41 that improved to 8.5 after the intervention further supported this. The mean adequacy ratio (MAR) showed a statistically significant improvement from baseline (0.77 +/- 0.48) to 1.02 +/- 0.66 after the intervention, indicating improvement of the adequacy of the overall diet to meet the daily requirements for this elderly community. CONCLUSIONS: The results of this study represent the first demonstration that food aid proved to be an effective short term nutrition intervention that improved the level of dietary diversity. Other food-based approaches or nutrition education activities should be implemented simultaneously to improve self sufficiency in such a community for long-term effectiveness in terms of nutrient inadequacy and dietary diversity. PMID- 19300865 TI - Unawareness of weight and height--the effect on self-reported prevalence of overweight in a population-based study. AB - OBJECTIVE: Self-reported height and weight are commonly used to estimate body mass index. The present study aims to identify the characteristics of participants who reported not to know their height or weight and to evaluate the consequence of these missing data on the estimates of the prevalence of body mass index categories. DESIGN AND PARTICIPANTS: We evaluated 1492 adults, aged 18-92 years, who were selected using random digit dialing. During a general interview, participants were asked to report their current height and weight and actual anthropometrical measurements were obtained as part of a subsequent physical examination. We classified as unaware subjects who reported not to know their height or weight. RESULTS: In this sample, 185 (12.4%) subjects reported not to know their height or weight (19.6% of females and 6.3% of males, p < 0.001). Women unaware of their weight or height were older, less educated, overweight or obese, never smoker and more frequently reported no regular physical exercise practice. In men, only ex-smokers were significantly less aware of their weight or height. In women, the prevalence of obesity using measured data was 25.3%, decreasing to 22.2% when considering only participants aware of their weight and height. Using self-reported data the obesity prevalence, in females, was 15.0%. CONCLUSION: A large proportion of adults, mainly women and older persons, reported not to know their weight and height and the exclusion of those participants results in differential errors depending on the actual values. The exclusion of participants unaware of their weight or height is an additional source of bias that decreases the sensitivity in detecting obesity and underestimates the real problem, namely in the older population. PMID- 19300866 TI - The association of thyroid stimulating hormone levels with cognitive function and depressed mood: the Rancho Bernardo study. AB - OBJECTIVE: To evaluate the association of thyroid stimulating hormone levels with cognitive function and depressed mood in a community-based sample. DESIGN: Cross sectional study. SETTING: Clinic visit in 1999- 2003. PARTICIPANTS: Community dwelling men (N=447) and women (N=663) aged 42-99 years. MEASUREMENT: Cognitive function was assessed with the Buschke-Fuld Selective Reminding Test, the Modified Mini-Mental State Examination, Trails B, and category fluency. Depressed mood was assessed with the Beck Depression Inventory (BDI). A fasting blood sample was obtained for thyroid stimulating hormone (TSH) measurement. RESULTS: Mean age was 73.6 +/- 10.0 in men and 74.3 +/- 10.4 in women. Mean TSH was 1.9 mu IU/ml in both sexes; 9.0% of men and 24% of women reported thyroid medication use. Mean BDI scores were 4.6 +/- 4.1 in men and 5.2 +/- 4.3 in women; 9% of men and 11% of women used antidepressants. Before and after adjustment for covariates or exclusion of participants taking thyroid hormones, no associations were observed between TSH and cognitive function (ps > 0.10). TSH was inversely associated with BDI (p=0.03) in men, but not women. CONCLUSIONS: Thyroid stimulating hormone level was unrelated to cognitive function in men and women, and was inversely associated with depressed mood in men only, possibly reflecting the greater use of both thyroid medications and antidepressants by women. PMID- 19300868 TI - Use of biomarkers in Alzheimer's trials. PMID- 19300867 TI - Sleep disorders and their impacts on healthy, dependent, and frail older adults. AB - BACKGROUND: Sleep disorders differ widely in the heterogeneous older adult population. Older adults can be classified into three groups based upon their overall level of disability: healthy, dependent, and frail. Frailty is an emerging concept that denotes older persons at increased risk for poor outcomes. OBJECTIVE: The aim of this consensus review is to describe the sleep disorders observed in healthy and dependent older adults and to discuss the potential sleep disorders associated with frailty as well as their potential consequences on this weakened population. METHODS: A review task force was created including neurologists, geriatricians, sleep specialists and geriatric psychiatrists to discuss age related sleep disorders depending on the three categories of older adults. All published studies on sleep in older adults on Ovid Medline were reviewed and 106 articles were selected for the purpose of this consensus. RESULTS: Many healthy older adults have complains about their sleep such as waking not rested and too early, trouble falling asleep, daytime napping, and multiple nocturnal awakenings. Sleep architecture is modified by age with an increased percentage of time spent in stage one and a decreased percentage spent in stages three and four. Insomnia is frequent and its mechanisms include painful medical conditions, psychological distress, loss of physical activity and iatrogenic influences. Treatments are also involved in older adults' somnolence. The prevalence of primary sleep disorders such as restless legs syndrome, periodic limb movements and sleep disordered breathing increases with age. Potential outcomes relevant to these sleep disorders in old age include mortality, cardiovascular and neurobehavioral co-morbidities. Sleep in dependent older adults such as patients with Alzheimer Disease (AD) is disturbed. The sleep patterns observed in these patients are often similar to those observed in non demented elderly but alterations are more severe. Nocturnal sleep disruption and daytime sleepiness are the main problems. They are the results of Sleep/wake circadian rhythm disorders, environmental, psychological and iatrogenic factors. They are worsened by other sleep disorders such as sleep disordered breathing. Sleep in frail older adults per se has not yet been formally studied but four axes of investigation should be considered: i) sleep architecture abnormalities, ii) insomnia iii) restless legs syndrome (RLS), iv) sleep disordered breathing. CONCLUSION: Our knowledge in the field of sleep disorders in older adults has increased in recent years, yet some groups within this heterogeneous population, such as frail older adults, remain to be more thoroughly studied and characterized. PMID- 19300869 TI - Imaging and biomarkers will be used for detection and monitoring progression of early Alzheimer's disease. PMID- 19300870 TI - Biomarkers for Alzheimer's disease trials--biomarkers for what? A discussion paper. PMID- 19300871 TI - Interpreting biomarker data in therapeutic trials. PMID- 19300872 TI - Biomarkers in clinical trials of Alzheimer disease (AD): what is expected from regulatory agencies? AB - Biomarkers are key for the different phases of drug development in Alzheimer Disease and in other fields. Traditionally the process of biomarker qualification has been slow, mostly exclusively academic. Regulators were mostly passive spectators. The Pharmaceutical innovation crisis of the end of XX century prompted the regulatory agencies and the governments from both sides of the Atlantic to initiate a number of programs to foster renovation, creativity and new paradigms. This created the opportunity for regulators to move from a passive position to a more active role, as facilitators. However, it is becoming apparent that there is room for even more central roles for regulators in the interactive consortia that are now in place. It is also becoming apparent that regulatory agencies are expected to become highly specialized hubs holding expert "know-how" not easily available elsewhere. PMID- 19300873 TI - How can we recognize "disease modification" effects? PMID- 19300874 TI - Use of biomarkers to select the target population for clinical trials in subjects with mild cognitive impairment. PMID- 19300875 TI - Molecular imaging markers in clinical trials in Alzheimer s disease. PMID- 19300876 TI - Neuro-imaging: structural and functional imaging. PMID- 19300877 TI - Structural imaging markers for therapeutic trials in Alzheimer's disease. PMID- 19300878 TI - Knowing the natural course of biomarkers in AD: longitudinal MRI, CSF and PET data. PMID- 19300879 TI - Interest of the new criteria for drug trials in AD. PMID- 19300880 TI - Use of CSF biomarkers in Alzheimer's disease clinical trials. PMID- 19300881 TI - What we have learned from the Myriad trials. PMID- 19300882 TI - What we have learned from the Xaliproden Sanofi-aventis trials. PMID- 19300883 TI - Challenges in the conduct of disease-modifying trials in AD: practical experience from a phase 2 trial of Tau-aggregation inhibitor therapy. PMID- 19300884 TI - Lessons learned in the use of volumetric MRI in therapeutic trials in Alzheimer's disease: the ALZHEMED (Tramiprosate) experience. PMID- 19300885 TI - Enrichment of MCI and early Alzheimer's disease treatment trials using neurochemical and imaging candidate biomarkers. AB - In the earliest clinical stages of Alzheimer's Disease (AD), when symptoms are mild, clinical diagnosis will still be difficult. AD related molecular mechanisms precede symptoms. Biological markers can serve as early diagnostic indicators, as markers of preclinical pathological change, e.g. underlying mechanisms of action (MoA). Hypothesis based candidates are derived from structural and functional neuroimaging as well as from cerebrospinal fluid (CSF) and plasma. Unbiased exploratory approaches e.g. proteome analysis or rater independent fully automated imaging post-processing methods yield novel candidates. Recent progress in the validation of core feasible imaging and neurochemical biomarkers for functions such as early detection, classification, progression and prediction of AD is summarized. Single core feasible biomarkers can already be used to enrich populations at risk for AD and may be further enhanced using distinct combinations. Some biomarkers are currently in the process of implementation as primary or secondary outcome variables into regulatory guideline documents, e.g. regarding phase II in drug development programs as outcome measures in proof of concept or dose finding studies. There are specific biomarkers available depending on the hypothesized mechanism of action of a medicinal product, e.g. impact on the amyloidogenic cascade or on tauhyperphosphorylation. Ongoing large scale international controlled multi-center trials will provide further validation of selected core feasible imaging and CSF biomarker candidates as outcome measures in early AD for use in phase III clinical efficacy trials. There is a need of rigorous co-development of biological trait- and statemarker candidates facilitated through planned synergistic collaboration between academic, industrial and regulatory partners. PMID- 19300886 TI - The validity of biomarkers as surrogate endpoints in Alzheimer's disease by means of the Quantitative Surrogate Validation Level of Evidence Scheme (QSVLES). AB - OBJECTIVE: To evaluate the validity of biomarkers that are currently being proposed as potential surrogate endpoints in AD clinical trials with the aid of the "Quantitative Surrogate Validation Level of Evidence Schema" (QSVLES) proposed by Lassere et.al. (1). PROCEDURE: A Pubmed literature search was conducted to identify AD biomarkers with SEP potential, and the QSVLES was applied to determine the extent of the SEP validity. RESULTS: MRI, PET and MRS measures attained a total validity score of 4, NAA/Cre a total score of 5, and cerebral blood flow (SPECT), Abeta , Tau and APP a total score of 2. None of these biomarkers could fall into the rank of Levels 1 or 2, reserved for SEPs, according to the QSVLES criteria. This was mainly attributed to the lack of sufficient evidence that was derived from high ranking studies (RCT, prospective observational studies). CONCLUSION: Though residing on SEPs as sole determinants of the benefit/risk ratio of AD medications seems to be pretty far, there could be certain cases where the use of SEPs may be beneficial, making efficient therapies available faster when there is a major public health interest involved. However, the potential risks of relying on invalid SEPs should not be underestimated and therefore the research on SEP validation and the development of specific validation guidance should be encouraged. The QSVLES, though not devoid of criticism, may be proposed as a starting point. PMID- 19300887 TI - Defining frailty--the Holy Grail of geriatric medicine. PMID- 19300888 TI - Frailty: defining and measuring of a concept. AB - Older, more vulnerable individuals are increasingly often described in the literature as being frail. Because frailty is often perceived as being undesirable and associated with high health risks, it is important to establish how we can predict, recognize, and treat frailty. Frailty is predisposed by advancing age in combination with physiological deterioration, especially a loss of muscle mass and bone density. Although the symptoms of frailty are diverse, the most common symptoms are a deterioration of activities of daily living (ADL), mobility, nutritional status, cognition, and endurance. The consequences of frailty are institutionalization, morbidity, and mortality. The main determinants of frailty are limitations in ADL, weight loss, diminished mobility or patterns of activity, lowered serum cholesterol level, and sensitivity to change. There is no gold standard for the measurement of frailty, and often studies use a combination of instruments. Although a couple of multidimensional instruments have been developed to measure frailty in its totality, the reliability and validity of these instruments have yet to be established. Successful interventions against frailty include increasing muscle strength through training and individualized recommendations made on the basis of an extensive geriatric assessment. PMID- 19300889 TI - You missed the 5th EUGMS Congress? All the things which you would have been able to learn! PMID- 19300890 TI - Regional origin and decrease of pain in patients with depressive symptoms under treatment with venlafaxine. AB - OBJECTIVE: Patient's language, tradition, conventions, and customs may all determine integration into a society and are also part of the doctor-patient relationship that influences diagnostic and therapeutic outcome. Language barrier and sociocultural disparity of Eastern and Southern European patients may hamper recovery from pain and depression compared to Middle European patients in Switzerland. METHODS: In a prospective naturalistic observational trial we investigated the influence of regional origin on treatment outcome in 420 pain sufferers with depressive symptoms from all over Switzerland who were treated with venlafaxine by 122 physicians in primary care. Physicians rated severity of depressive symptoms using the clinical global impression severity scale and pain intensity by means of visual analogue scales. We hypothesized that in Eastern and Southern European patients the magnitude of pain reduction under treatment with venlafaxine is less compared to Middle European patients. RESULTS: Three months after study entry, Middle European patients were found to profit more from treatment with venlafaxine in terms of severity of depression and pain intensity than patients from Eastern Europe and Southern Europe. CONCLUSION: Regional origin may contribute to the magnitude of pain reduction in patients with depressive symptoms under treatment with venlafaxine. Our results provide a rational for care provider educational programs aimed at improving capacities in treating patients from different regional origin with psychosomatic complaints such as depression and comorbid pain. PMID- 19300891 TI - Morphological changes of injected calcium phosphate cement in osteoporotic compressed vertebral bodies. AB - SUMMARY: This study was undertaken to investigate the radiologic and clinical outcomes of vertebroplasty with calcium phosphate (CaP) cement in patients with osteoporotic vertebral compression fractures. The morphological changes of injected CaP cement in osteoporotic compressed vertebral bodies were variable and unpredictable. We suggest that the practice of vertebroplasty using CaP should be reconsidered. INTRODUCTION: Recently, CaP, an osteoconductive filler material, has been used in the treatment of osteoporotic compression fractures. However, the clinical results of CaP-cement-augmented vertebrae are still not well established. The purpose of this study is to assess the clinical results of vertebroplasty with CaP by evaluating the morphological changes of CaP cement in compressed vertebral bodies. METHODS: Fourteen patients have been followed for more than 2 years after vertebroplasty. The following parameters were reviewed: age, sex, T score, compliance with osteoporosis medications, visual analog scale score, compression ratio, subsequent compression fractures, and any morphological changes in the filler material. RESULTS: The morphological changes of injected CaP included reabsorption, condensation, bone formation (osteogenesis), fracture of the CaP solid hump, and heterotopic ossification. Out of 14 patients, 11 (78.6%) developed progression of the compression of the CaP-augmented vertebral bodies after vertebroplasty. CONCLUSIONS: The morphological changes of the injected CaP cement in the vertebral bodies were variable and unpredictable. The compression of the CaP-augmented vertebrae progressed continuously for 2 years or more. The findings of this study suggest that vertebroplasty using CaP cement should be reconsidered. PMID- 19300892 TI - Chronic obstructive pulmonary disease is associated with osteoporosis and low levels of vitamin D. AB - We did a cross-sectional analysis of chronic pulmonary obstructive disease (COPD) patients without chronic use of systemic glucocorticoids (CUG). Osteoporosis was found in 51% and bone mineral density (BMD) was correlated with severity of disease. Low levels of vitamin D were found in 94%. All COPD patients may benefit from vitamin D supplementation and screening for low BMD. INTRODUCTION: Patients with chronic pulmonary obstructive disease have low bone mineral density, caused by chronic use of systemic glucocorticoids and hypovitaminosis D. However, patients without CUG may also have low BMD. METHODS: We performed a cross sectional analysis in 49 patients (21 men, 28 postmenopausal women), with COPD without CUG, from Brazil (25 degrees 25' S). Several markers of bone metabolism were measured, plus BMD. Osteoporosis risk factors and history of fractures were investigated. Respiratory function was assessed by venous gasometry, spirometry, and oximetry. BMD results were compared to those of 40 healthy non-smokers controls. RESULTS: COPD patients had lower BMD at all sites (p < 0.01). Osteoporosis was observed in 51%. BMD independently correlated with stage of disease (lumbar spine, R = 0.38, p = 0.01; total femur, R = 0.36, p = 0.01; femoral neck, R = 0.40, p < 0.01). Ninety-four percent had low levels of vitamin D (<30 ng/mL) and 67% had secondary hyperparathyroidism. Vitamin D was correlated with oxygen saturation (R = 0.36, p = 0.01), with lower levels in those with saturation <88% (p = 0.01). CONCLUSION: Patients with COPD without CUG have increased risk for osteoporosis. Such patients have hypovitaminosis D, which is correlated with the severity of disease. Screening for low BMD and vitamin D supplementation may be warranted to all COPD patients. PMID- 19300893 TI - A recurrent mutation c.617G>A in the ACVR1 gene causes fibrodysplasia ossificans progressiva in two Chinese patients. AB - Fibrodysplasia ossificans progressiva (FOP; OMIM 135100) is a rare heritable disorder of connective tissue characterized by congenital malformations of the great toes and recurrent episodes of painful soft tissue swelling that lead to heterotopic ossifications. Recent studies have shown that the ACVR1 (activin A receptor, type I; OMIM 102576) gene, which encodes the BMP type I receptor protein, is responsible for this disease. We observed two Chinese patients who suffered from progressive pain and ankylosis of major joints with congenital bilateral hallus valgus malformation, neck stiffness, and several posttraumatic ossified lesions on the head and dorsum. Both patients were diagnosed as having FOP. This study aimed to investigate the ACVR1 gene mutation in Chinese FOP patients. Direct sequence analysis of genomic DNA and restriction enzyme digestion demonstrated the presence of a single heterozygous c.617G>A (p.R206H) mutation in the ACVR1 gene in both patients. This mutation is first reported in Chinese patients with FOP and it was de novo in both affected families. PMID- 19300894 TI - Extra- and intramuscular nerves distributions of the triceps surae muscle as a basis for muscle resection and botulinum toxin injections. AB - PURPOSE: To compare the distribution of extramuscular nerve branches with their intramuscular ramifications in the triceps surae muscle, thus providing anatomical substantiation for the topography of muscle resection and botulinum toxin injections. METHODS: Dissection and modified Sihler's staining of 18 whole mount human cadaveric specimens. RESULTS: The distance between the areas with the highest extramuscular branch density and the area of densest intramuscular arborization in gastrocnemius and soleus muscles is approximately 10% of the calf length. This finding should be taken into consideration during nerve blocking and botulinum toxin injections for the treatment of spasticity. Intramuscular nerve arborization patterns make it possible to outline neuromuscular segments in the gastrocnemius and soleus muscles. CONCLUSIONS: Surgical or therapeutic interventions in areas of high extramuscular and intramuscular nerve density can increase the efficacy and safety of botulinum toxin injections and neurotomy. Intramuscular nerve branching patterns should be taken into consideration during triceps surae resection. PMID- 19300895 TI - Efficacy of preoperative endoscopic nasobiliary drainage for hilar cholangiocarcinoma. AB - BACKGROUND/PURPOSE: Although percutaneous transhepatic biliary drainage has previously been recommended as a primary preoperative step, endoscopic nasobiliary drainage (ENBD) is prevalent as an alternative procedure. Few reports assess the efficacy and safety of ENBD in a substantial patient cohort. METHODS: Of 116 patients with hilar cholangiocarcinoma who underwent surgery, 62 (43 men and 19 women, median age 69 years) underwent preoperative ENBD. After classification of lesions according to Bismuth-Corlette (B-C) criteria, we evaluated efficacy and safety with respect to B-C type. RESULTS: Patients were classified as B-C types I (n = 5), II (n = 21), IIIa (n = 23), IIIb (n = 5), and IV (n = 8). Preoperative single ENBD was effective in 46/62 patients (74%) including 5/5 (100%) B-C type I, 20/21 (94%) type II, 16/23 (70%) type IIIa, 4/5 (80%) type IIIb, and 1/8 (13%) type IV. Sixteen cases (26%) required additional drainages with ENBD or endoscopic biliary stenting (EBS) in 8/16 (50%), and with PTBD in 8/16 (50%). Mild acute pancreatitis (n = 1, 2%), segmental cholangitis (n = 2, 3%), and acute cholangitis with catheter obstruction (n = 7, 11%) occurred with ENBD. CONCLUSIONS: Preoperative single ENBD in the future remnant lobe is effective treatment for B-C type I-III hilar cholangiocarcinoma. Preoperative ENBD was rarely complicated with segmental cholangitis. PMID- 19300896 TI - Surgical resection versus radiofrequency ablation for small hepatocellular carcinomas within the Milan criteria. AB - BACKGROUND/PURPOSE: It has been reported that hepatic resection may be preferable to other modalities for the treatment of small hepatocellular carcinomas (HCCs), by contributing to improved overall and disease-free survival. Ablation techniques such as radiofrequency ablation (RFA) have also been used as therapy for small HCCs; however, few studies have compared the two treatments based on long-term outcomes. The effectiveness of hepatic resection and RFA for small nodular HCCs within the Milan criteria were compared. METHODS: A retrospective cohort study was performed with 278 consecutive patients who underwent curative hepatic resection (n = 123) or initial RFA percutaneously (n = 110) or surgically (thoracoscopic-, laparoscopic-, and open-approaches; n = 45) for HCC. The selection criteria for treatment were based on uniform criteria. Mortality related to therapy and 3- and 5-year overall and disease-free survivals were analyzed. RESULTS: The model for endstage liver disease (MELD) scores for all patients in the series were less than 13. There were no therapy-related mortalities in either the hepatic resection or RFA groups. The incidence of death within 1 year after therapy (1.6 and 1.9%, respectively) was similar in the hepatic resection and RFA groups. The group that underwent hepatic resection showed a trend towards better survival (P = 0.06) and showed significantly better disease-free survival (P = 0.02) compared with the RFA group, although differences in liver functional reserve existed. The advantage of hepatic resection was more evident for patients with single tumors and patients with grade A liver damage. In contrast, patients with multinodular tumors survived longer when treated with RFA, regardless of the grade of liver damage. Further analysis showed that surgical RFA could potentially have survival benefits similar to those of hepatic resection for single tumors, and that surgical RFA had the highest efficacy for treating multinodular tumors. CONCLUSIONS: In patients with small HCCs within the Milan criteria, hepatic resection should still be employed for those patients with a single tumor and well-preserved liver function. RFA should be chosen for patients with an unresectable single tumor or those with multinodular tumors, regardless of the grade of liver damage. In order to increase long-term oncological control, surgical RFA seems preferable to percutaneous RFA, if the patient's condition allows them to tolerate surgery. PMID- 19300897 TI - Independent factors associated with depression in type 1 diabetes mellitus. AB - Diabetes has been associated with depression since Thomas Willis' work in 1684 (Rubin and Peyrot in Diabetes Metab Rev 18:173-175, 2002). The aim of this study is to identify social and clinical factors independently associated with depression in individuals with type 1 diabetes. We carried out a descriptive transversal study with 110 type 1 diabetes patients, administered a questionnaire and obtained demographical and diabetes-related data (number of years from diagnosis, initial admission at diagnosis, glycated hemoglobin, number of complications, insulin dose, number of insulin injections per day, admission for ketoacidosis or hypoglycemia at diagnosis, and specific diabetes complications such as nephropathy, retinopathy, peripheral neuropathy, coronariopathy, and amputation). Depressive symptoms were quantified using the Hamilton Score. We used T tests to investigate potential relations between the covariates and depression (Hamilton score). We concluded the following: as few as 10% of our patients had glycated hemoglobin under 7%; women had more symptoms of depression, and there are four independent factors associated with depression in individuals with type 1 diabetes mellitus: age, Graffar score, admission for ketoacidosis, and insulin dose. PMID- 19300898 TI - Universal versus selective screening for the detection, control and prognosis of gestational diabetes mellitus in Argentina. AB - In all, 1,702 unselected pregnant women from the city of La Plata were tested for gestational diabetes mellitus (GDM) and evaluated to determine GDM prevalence and risk factors. In women with GDM, we evaluated compliance with guidelines for GDM management, and perinatal complications attributable to GDM. GDM prevalence was 5.8%, and its risk factors were pre-gestational obesity, previous hyperglycaemia, age > 30 years, previous GDM (and its surrogate markers). In primi-gravida (PG) subjects, GDM was equally prevalent in the presence (4.2%) or absence (4.0%) of risk factors. In multi-gravida (MG) women, although risk factors doubled the prevalence of GDM (8.6%), in the absence of risk factors GDM prevalence was similar to that of PG women (3.9%). Half of all women with GDM received inadequate post-diagnosis obstetric control, and this induced a fourfold increase in infant perinatal complications. In conclusion, all non-hyperglycaemic 24-28 week pregnant women should be tested for GDM, although particular attention must be paid to MG women with risk factors. PMID- 19300899 TI - Pantalone et al.: the risk of developing coronary artery disease or congestive heart failure, and overall mortality, in type 2 diabetic patients receiving rosiglitazone, pioglitazone, metformin, or sulfonylureas: a retrospective analysis. PMID- 19300900 TI - Study on management of pediatric migraine by general practitioners in northern France. AB - The study explored the awareness of the Haute Autorite de Sante (High Health Authority, HAS) guidelines for migraine management in children among a random sample of 100 general practitioners (GPs) dichotomised in an urban and a rural group. A questionnaire conducted by phone included questions on knowledge of pediatric migraine acute treatment and preventive therapy, referral to a child neurologist as well as GPs awareness of HAS recommendations in general. Although 45% of GPs argued they were prescribing ibuprofen as first-line abortive drug, only 3% were aware of the recommended dose. Only 48% of GPs were agreeing to initiate preventive therapy. Fifty percent of GPs stated that they knew HAS guidelines but only 24% stated that they had read them. The only significant difference between urban and rural GPs concerned the initiation of preventive therapy. Continuing educational programmes on the implementation of pediatric migraine guidelines is strongly needed. PMID- 19300901 TI - Image guided percutaneous trans-pedicular screw fixation of the thoracic spine. A clinical evaluation. AB - OBJECTIVE: We report our preliminary experience with a minimally invasive image guided percutaneous trans-pedicular fixation technique of the thoracic spine in patients with significant co-morbidity. This study aims to demonstrate the feasibility and efficacy of the presented operative technique as well as to detect potential drawbacks. METHOD: A navigated percutaneous application of trans pedicular screws was performed in 14 patients with radiological validated instability of the thoracic spine and significant medical co-morbidity. Due to poor bone quality, vertebroplasty of the affected levels had to be performed in nine patients. The levels involved were (T9-12) in ten patients, the middle (T5 8) in two patients and the upper thoracic (T1-4) segments in two others. VAS scores for local back pain were used to assess clinical outcome. RESULTS: A total of 56 screws were inserted. There was no additional morbidity associated with the procedure. Post-operative CT scans and plain X-rays were obtained in all patients. In 2 (3.5%) medial penetration of the pedicle border occurred without neurological sequelae for the patient. Reduction of VAS scores regarding back pain during the first post-operative week was noted. Follow up ranged between 6 months and 12 months. CONCLUSION: Navigated percutaneous trans-pedicular fixation of the thoracic spine is feasible and can be performed safely in patients where open surgery is of significant risk. Pre-operative planning is essential in order to avoid intra-operative complications with the instrumentation system. PMID- 19300902 TI - Vascular response to the vasa vasorum in a carotid artery dissecting aneurysm. AB - Collateralization of a hemodynamically compromised internal carotid artery (ICA) by the vasa vasorum is rare. We report the serial radiologic findings of an apparent hypertrophied vasa vasorum of the cervical ICA, which was secondary to arterial dissection. PMID- 19300903 TI - Trigonal cavernous angioma: a short illustrated review. AB - PURPOSE: Intraventricular cavernomas are rare. Even more rare are those presenting in the trigone of the lateral ventricles. METHODS: We performed a search of the literature of the last 30 years and identified all cases of intraventricular cavernous angiomas. Trigonal cavernomas were separately identified and analysed. Our search yielded a total of 13 trigonal cavernomas. RESULTS: Of a total of 61 intraventricular cases, 13 were located in the trigone of the lateral ventricles. The most prominent presenting symptom was intracranial hypertension (68.9%), followed by seizures (18.2%) and hemorrhage (13.1%).The literature review revealed a trend of intraventricular cavernomas to present with intracranial hypertension rather than seizures or focal neurologic deficit, unlike their intraparenchymal counterparts. We feel that this difference has received little attention in the international literature. We discuss a possible pathogenetic mechanism for the presence of intracranial hypertension and address different aspects of diagnosis and treatment of this benign lesion. CONCLUSIONS: Trigonal cavernomas are benign lesions that have an excellent outcome after radical excision. Symptoms and signs of intracranial hypertension and hydrocephalus may be the prominent initial presentation of this rare ailment. PMID- 19300904 TI - Isolated subacute tuberculous spinal epidural abscess of the cervical spine: a brief report of a special case. AB - A tuberculous spinal epidural abscess is seen rarely as a late complication of Pott's disease or in immunocompromised patients. Such abscesses in isolation are rare indeed and very uncommon in the developed and developing world. We report a patient with an isolated subacute tuberculous spinal epidural abscess without disc or vertebral involvement and no primary focus or risk factors associated with the development of spinal tuberculosis. PMID- 19300905 TI - Membrane functional organisation and dynamic of mu-opioid receptors. AB - The activation and signalling activity of the membrane mu-opioid receptor (MOP-R) involve interactions among the receptor, G-proteins, effectors and many other membrane or cytosolic proteins. Decades of investigation have led to identification of the main biochemical processes, but the mechanisms governing the successive protein-protein interactions have yet to be established. We will need to unravel the dynamic membrane organisation of this complex and multifaceted molecular machinery if we are to understand these mechanisms. Here, we review and discuss advances in our understanding of the signalling mechanism of MOP-R resulting from biochemical or biophysical studies of the organisation of this receptor in the plasma membrane. PMID- 19300906 TI - Processing of peptide and hormone precursors at the dibasic cleavage sites. AB - Many functionally important cellular peptides and proteins, including hormones, neuropeptides, and growth factors, are synthesized as inactive precursor polypeptides, which require post-translational proteolytic processing to become biologically active polypeptides. This is achieved by the action of a relatively small number of proteases that belong to a family of seven subtilisin-like proprotein convertases (PCs) including furin. In view of this, this review focuses on the importance of privileged secondary structures and of given amino acid residues around basic cleavage sites in substrate recognition by these endoproteases. In addition to their participation in normal cell functions, PCs are crucial for the initiation and progress of many important diseases. Hence, these proteases constitute potential drug targets in medicine. Accordingly, this review also discusses the approaches used to shed light on the cleavage preference and the substrate specificity of the PCs, a prerequisite to select which PCs are promising drug targets in each disease. PMID- 19300907 TI - The long journey of actin and actin-associated proteins from genes to polysomes. AB - During gene expression, multiple regulatory steps make sure that alterations of chromatin structure are synchronized with RNA synthesis, co-transcriptional assembly of ribonucleoprotein complexes, transport to the cytoplasm and localized translation. These events are controlled by large multiprotein complexes commonly referred to as molecular machines, which are specialized and at the same time display a highly dynamic protein composition. The crosstalk between these molecular machines is essential for efficient RNA biogenesis. Actin has been recently proposed to be an important factor throughout the entire RNA biogenesis pathway as a component of chromatin remodeling complexes, associated with all eukaryotic RNA polymerases as well as precursor and mature ribonucleoprotein complexes. The aim of this review is to present evidence on the involvement of actin and actin-associated proteins in RNA biogenesis and propose integrative models supporting the view that actin facilitates coordination of the different steps in gene expression. PMID- 19300908 TI - The protein arginine methyltransferase family: an update about function, new perspectives and the physiological role in humans. AB - Information about the family of protein arginine methyltransferases (PRMTs) has been growing rapidly over the last few years and the emerging role of arginine methylation involved in cellular processes like signaling, RNA processing, gene transcription, and cellular transport function has been investigated. To date, 11 PRMTs gene transcripts have been identified in humans. Almost all PRMTs have been shown to have enzymatic activity and to catalyze arginine methylation. This review will summarize the overall function of human PRMTs and include novel highlights on each family member. PMID- 19300909 TI - Chemopreventive efficacy of gallic acid, an antioxidant and anticarcinogenic polyphenol, against 1,2-dimethyl hydrazine induced rat colon carcinogenesis. AB - Colon cancer is a major cause of morbidity and mortality in developed and developing countries and its etiology is known to be a combination of hereditary, environmental, dietary factors and lack of physical activity. Chemoprevention offers a novel approach to control the incidence of colon cancer. Gallic acid (GA) is a polyphenol widely present in tea and other plants which is popularly used in the traditional medicine of China. The present study was to evaluate the efficacy of GA supplementation on tissue lipid peroxidation and antioxidant defense system in 1,2-dimethyhydrazine (DMH) induced colon carcinogenesis in male Wistar rats. The rats were assorted into six groups, viz., group1 control rats received modified pellet diet; group 2 rats received GA (50 mg/kg body weight) orally along with modified pellet diet; group 3 rats received DMH (20 mg/kg body weight) subcutaneously once a week for the first 15 weeks; groups 4, 5 and 6 rats received GA along with DMH during the initiation, post- initiation stages and the entire period of study respectively. All the rats were sacrificed at the end of 30 weeks and the tissues were evaluated biochemically. We observed decreased lipid peroxidation (LPO) products such as thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (LOOH) and conjugated dienes (CD) and diminished levels of antioxidants such as superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione reductase (GR) and glutathione peroxidase (GPx) in the tissues of DMH treated rats, which were elevated significantly on GA supplementation. Moreover, enhanced activity of ascorbic acid and alpha tocopherol levels were also observed in DMH alone treated rats which were significantly reduced on GA supplementation. Our results suggest that GA could exert a significant chemopreventive effect on DMH induced colon carcinogenesis. PMID- 19300910 TI - Upregulation of LRIG1 suppresses malignant glioma cell growth by attenuating EGFR activity. AB - Activated epidermal growth factor receptor (EGFR) has emerged as an important therapeutic target for a variety of solid tumors, particularly malignant gliomas. A recently discovered transmembrane glycoprotein, LRIG1, antagonizes the activity of epidermal growth factor receptor family receptor tyrosine kinases and acts as a negative feedback loop of EGFR and proposed tumor suppressors. The aim of this study was to investigate the impact of LRIG1 on the biological features of glioma cells and the possible mechanisms of enhanced apoptosis induced by upregulation of LRIG1. We observed that the expression of LRIG1 was decreased, while the expression of EGFR was increased in the majority of astrocytomas, and the ratio of EGFR/LRIG1 was increased by sixfold in tumors versus corresponding non neoplastic tissue. Upregulation of LRIG1, followed by a decrease of EGFR on the cytomembrane of the cells, induced cell apoptosis and cell growth inhibition, and further reversed invasion in glioma cell lines and primary glioma cells. Our study now clearly indicates that LRIG1 indeed affects cell fate and biology behaviors of the cells in vitro by inhibiting phosphorylation of downstream MAPK and AKT signaling pathway, and the elevated release level of caspase-8 might contribute to the enhanced apoptosis in LRIG1 transfected glioma cells. Taken together, these findings provide us with an insight into LRIG1 function, and we conclude that LRIG1 evolved in gliomas as a rare feedback negative attenuator of EGFR and could offer a novel therapeutic target to treat patients with malignant gliomas. PMID- 19300911 TI - Dose-dependent RNAi-mediated geminivirus resistance in the tropical root crop cassava. AB - Cassava mosaic disease is a major constraint for cassava production in Africa, resulting in significant economic losses. We have engineered transgenic cassava with resistance to African cassava mosaic virus (ACMV), by expressing ACMV AC1 homologous hairpin double-strand RNAs. Transgenic cassava lines with high levels of AC1-homologous small RNAs have ACMV immunity with increasing viral load and different inoculation methods. We report a correlation between the expression of the AC1-homologous small RNAs and the ACMV resistance of the transgenic cassava lines. Characterization of the small RNAs revealed that only some of the hairpin derived small RNAs fall into currently known small interfering RNA classes in plants. The method is scalable to stacking by targeting multiple virus isolates with additional hairpins. PMID- 19300912 TI - [Fast tracking in pediatric surgery]. AB - Fast track protocols are justified for several pediatric surgical procedures. Particular operations on neonates and babies require specialized postoperative care. In these very young patients, no benefit can be expected from rigid fast track concepts. Many workflows in pediatric surgery have already been optimized with a view toward very short hospital stay. The published experiences with fast track protocols in pediatric surgery show that the method is feasible and safe. Complication and readmission rates are very low. Perioperative adjustments, shorter fasting periods, and the use of new analgetics are desirable in pediatric fast track protocols. Particularly for children, customizing protocols to the individual patients' requirements must be guaranteed. PMID- 19300913 TI - [Shulman's syndrome (eosinophilic fasciitis)]. AB - Eosinophilic fasciitis is a rare disease characterized by edema, painful indurations, and progressive muscle weakness. Mainly the extremities are involved. We report on a 22-year-old woman with eosinophilic fasciitis presenting with progressive muscle weakness of both hands and feet and a reduced general condition. She showed symmetrical and firm swelling of the extremities with painful restriction of joint movement. Systemic treatment with glucocorticosteroids as well as physiotherapy and manual lymphatic drainage led to continuous improvement of her symptoms. The differentiation from other diseases, such as systemic scleroderma, eosinophilia-myalgia syndrome, and pseudoscleroderma, might be difficult at the beginning of the disease. The gold standard for diagnosis is--as was done in our case--a deep skin-to-muscle biopsy. Further imaging, especially magnetic resonance imaging, can support the diagnostic procedure. PMID- 19300914 TI - Abstracts of the Society of Behavioral Medicine 2009 Annual Meeting. April 22-25, 2009. Montreal, Quebec, Canada. PMID- 19300915 TI - Excitotoxicity in rat's brain induced by exposure of manganese and neuroprotective effects of pinacidil and nimodipine. AB - Manganese (Mn) is an essential trace element for humans. However, manganism would be caused by excessive Mn. The mechanisms underlying excitotoxicity induced by manganism are poorly understood. As it is known to us, glutamate (Glu) is the most prevalent excitatory neurotransmitter. To determine the possible role of dysfunction of Glu transportation and metabolism in Mn-induced excitotoxicity, the rats were ip injected with different dose of MnCl(2) (0, 50, 100, and 200 micromol/kg), the levels of Mn and activities of GS, PAG, Na(+)-K(+)-ATPase, and Ca(2+)-ATPase in striatum were investigated. In addition, effect of 20.38 micromol/kg pinacidil (K(+) channel opener) or 2.4 micromol/kg nimodipine (Ca(2+) channel blocker) were studied at 200 micromol/kg MnCl(2). With dose-dependent inhibition of GS, Na(+)-K(+)-ATPase, and Ca(2+)-ATPase activities, increase of Mn levels and PAG activity were observed. Further investigation indicated that pre treatment of pinacidil or nimodipine reversed toxic effect of MnCl(2) significantly. These results suggested that MnCl(2) could induce dysfunction of Glu transportation and metabolism by augmenting the excitotoxicity dose dependently; pinacidil and nimodipine might antagonize manganese neurotoxicity. PMID- 19300916 TI - Roles of organic acids and nitrate in the long-distance transport of cobalt in xylem saps of Alyssum murale and Trifolium subterraneum. AB - Roles of organic acids and nitrate in the long-distance transport of cobalt (Co) in xylem saps of hyperaccumulator Alyssum murale and non-hyperaccumulator Trifolium subterraneum were studied under hydroponic conditions. Organic acids (oxalic, malic, malonic, citric, and fumaric) and nitrate in xylem sap samples were separated and determined simultaneously by reversed-phase high performance liquid chromatography after solid-phase extraction with nanosized hydroxyapatite. Results indicated that Co treatment significantly increased the concentrations of xylem oxalic and malic acids for the hyperaccumulator A. murale compared to the control but significantly decreased the concentrations of xylem nitrate and malonic acid; concentrations of citric acid in xylem sap samples did not show significant difference between the control and Co treatments. By analyzing the relationship between the concentrations of organic acids, nitrate, and concentrations of Co in xylem saps, it could be concluded that oxalic and malic acids in xylem saps seemed to participate in the long-distance Co translocation process, and citric acid did not relate to the xylem Co transport of A. murale and T. subterraneum. Our work might be very useful for understanding the mechanism of long-distance transport of heavy metals in hyperaccumulator. PMID- 19300917 TI - Serum selenium level in healthy Koreans. AB - Of trace elements in the serum of living organisms, selenium (Se) is an essential mineral and plays the role of an antioxidant as selenoproteins protecting the organism against oxidative damage induced by hydrogen peroxide, other lipid hydroperoxides, and their derivatives. The aim of this study was to determine the mean serum Se levels in healthy Korean volunteers (50 males and 50 females) by using an inductively coupled plasma-mass spectrometry method. The samples were collected at the Health Promotion Centre of Kangnam St. Mary's Hospital, College of Medicine, the Catholic University of Korea, Kangnam District, Seoul in accordance with procedures approved by the Institutional Review Board of the Catholic University of Korea. The mean serum Se level in healthy subjects was 112.05 +/- 30.42 microg/l. For gender, it was 120.81 +/- 27.37 microg/l for females and 103.29 +/- 31.05 microg/l for males. From the study result, there was a significant difference between the mean Se concentrations of gender groups (p = 0.0035). Also, the study indicated no effect of age on Se levels (p > 0.05) in the healthy individuals. PMID- 19300952 TI - Potentials and limitations of current and emerging immunotherapies. PMID- 19300953 TI - Pathophysiology of multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS). Both genetic and environmental causes for MS have been suggested. Recent genome-wide association studies revealed new susceptibility alleles for MS besides the HLA complex that are all related with immune functions. Whereas there is little evidence to support a purely environmental trigger for the disease in the sense of an infectious agent, the autoimmune hypothesis of MS is well established. Myelin antigen-specific CD4+ T cells become activated in the peripheral immune compartment, cross the blood brain barrier and trigger the disease. Here, current concepts of the commitment of T cells to pro-inflammatory effector T helper cell lineages including Th17 cells that appear to be important inducers of organ-specific autoimmunity will be discussed. PMID- 19300954 TI - Animal models of multiple sclerosis for the development and validation of novel therapies - potential and limitations. AB - Various types of experimental autoimmune encephalomyelitis (EAE) reflect some of the pathogenetic, clinical, and therapeutic features of the different forms of multiple sclerosis (MS), thereby, providing some, albeit limited, insight into the molecular and cellular basis of the human disease. Specific questions of MS therapy including the search for new therapeutic targets and strategies and their validation require investigations in different available EAE models. A survey is given of experimental therapeutic approaches that are currently under study with the most promising examples of monoclonal antibodies, gene therapy, stem cell transplantation and orally applied small molecular weight disease-modifying drugs. Reasons for therapy failure and adverse side-effects of some experimental trials are discussed. Precaution is advised, if results of new experimental approaches are translated into clinical practice. PMID- 19300955 TI - Immunomodulatory treatment strategies in multiple sclerosis. AB - Multiple sclerosis (MS) represents the prototypic inflammatory autoimmune disorder of the central nervous system and the most common cause of neurological disability in young adults. The mainstays in the immunomodulatory therapy of MS are currently interferon beta and glatiramer acetate, both of which have proven to be clinically and paraclinically effective. Current clinical evidence indicates that treatment should be initiated as early as possible. In this review we summarize available data from clinical studies on clinical efficacy of immunomodulatory drugs for treating patients with multiple sclerosis. PMID- 19300956 TI - Immunosuppression in clinical practice: approaches to individualized therapy. AB - Despite novel immunoactive agents, immunosuppressants still play a considerable role in the treatment of MS, especially in rapidly progressive cases. Given the limited tolerability and potentially severe side effects of most immunosuppressive drugs, identification of patients with a favorable benefit-risk profile is essential. A narrow therapeutic index, with sometimes high interindividual variability in terms of response and side effects may partially be explained by genetic factors affecting different metabolic pathways. Here, we will review practical aspects in the clinical use of immunosuppressants in MS and discuss approaches to individualized treatment schemes, including novel pharmacogenetic strategies. PMID- 19300957 TI - Monoclonal antibodies in the therapy of multiple sclerosis: an overview. AB - With the generation of monoclonal antibodies (mAbs), a new therapeutical concept has gained importance. MAbs aim against selective antigens and so have changed our treatment strategies from non-specific to specific. Four therapeuticals have gained importance in the therapy of multiple sclerosis (MS): One has already been approved for therapy (natalizumab), whereas the other three are either in clinical trials or are about to enter phase III studies. Currently, two phase III studies that evaluate the efficacy of alemtuzumab have begun with recruitment (MS CARE I and II). Another mAb (daclizumab) under study is directed to the interleukin-2alpha chain (CD25). Results of clinical trials are promising by reporting reduction of relapses and progression in relapsing remitting and secondary progressive MS accompanied by reduction of new lesions in magnetic resonance imaging. A multicenter randomized controlled trial of daclizumab in MS is going to be initiated. Trials with a humanised antibody directed against the cell surface molecule CD20 are under development. Although the future will emphasise this trend to mAbs, the risks should not be ignored as has been shown in recent news. Still, mAbs have the possibility to revolutionise therapeutical concepts in the treatment of immune-mediated diseases, and will therefore be a useful addition to current therapeutic concepts. PMID- 19300958 TI - Corticosteroids and plasma exchange in multiple sclerosis. AB - Corticosteroids (CS) remain a mainstay of treatment for relapses in multiple sclerosis (MS) and optic neuritis. Currently, there is not enough evidence that long-term corticosteroid treatment delays progression of long-term disability in patients with MS. Likewise, it is unclear whether there are, in fact, true differences among the various CS agents, doses, and their applications in specific pulse and tapering regimens.In some patients suffering from severe steroid-resistant relapses, the clinical response to CS treatment may be insufficient. Such patients may obtain clinical benefit from subsequent plasma exchange (PE). PE is increasingly considered as an individual treatment decision in patients with severe relapses not properly responding to CS. Because of the lack of appropriate studies, PE is not recommended as a permanent disease modifying strategy in MS patients. PMID- 19300959 TI - Stem cell transplantation in multiple sclerosis. AB - Following promising results in animal studies showing that immunosuppression and consecutive allogeneic bone marrow transplantation has the potential to significantly reduce autoimmunity, emerging data is supporting a benefit in patients with multiple sclerosis (MS) and other autoimmune disorders not responding to approved therapies. Until today, results on over 400 cases have been reported by the European Blood and Marrow Transplantation Group (EBMT), many of them with a favorable outcome. However, results of randomized, controlled clinical trials are missing. Furthermore, with upcoming new treatment compounds that to some extent act via lymphoablative properties, it remains essential to better select those patients who might profit most from stem cell therapy based on a justifiable benefit-to-risk ratio. PMID- 19300960 TI - Monitoring of multiple sclerosis immunotherapy: from single candidates to biomarker networks. AB - Applying microarray technology to identify new diagnostic and prognostic markers in peripheral blood cells (PBC) after therapeutic intervention opens great perspectives regarding patient subclassification. Three recombinant products of the pleiotropic agent interferon beta (rIFN-beta) are available for disease modifying therapy of relapsing remitting multiple sclerosis (RRMS), a complex inflammatory autoimmune disease that targets the central nervous system. They differ according to formulation, route of administration and dosage regimens. The currently, only partially understood mechanism of action of injected rIFN-beta into human organisms needs provision with accessory key molecules; in addition, the significance of established clinical IFN-beta response criteria that distinguish responding from non-responding patients remain unclear.With respect to these major questions, we discuss promising candidates on the gene transcription level, attained from scientific MS literature that included a longitudinal aspect. Reviewed studies were in part carried out with distinct gene interrogating platforms (GeneArrays; RT-PCR), settings (in vitro; ex vivo), and study designs (drug formulations and regimen; inclusion criteria and clinical endpoints), hampering meaningful meta-analysis. Nevertheless, PBC from therapy naive MS patients, rIFN-beta treated MS patients, and healthy controls served to characterize facets of both the disease and its treatment. Hence, the field of MS transcriptomics in immunomodulatory therapy is (by far) not adequately understood and should be embedded into systems biology disciplines, yielding multi-layer analyses that deliver timely identification of MS subjects who will profit from applied rIFN-beta therapy. PMID- 19300961 TI - alpha4-Integrin antagonism with natalizumab: effects and adverse effects. AB - Based on the results of two phase III clinical trials, the humanized recombinant monoclonal antibody natalizumab was approved for the treatment of relapsing forms of multiple sclerosis (MS). Since its initial approval in November 2004, it has been announced that six patients who received natalizumab in the context of clinical studies acquired an infection with the human polyoma virus JC and were diagnosed with progressive multifocal leukoencephalopathy (PML). Two of these individuals had a fatal outcome. Our groups recently showed that natalizumab therapy results in a reduction of CD4(+) T cells within the cerebrospinal fluid (CSF) that is ten-fold more pronounced than the reduction in the number of CD8(+) T lymphocytes. Interestingly, it appears that the effect of natalizumab on cell numbers in the CSF persists for at least 6 months after cessation of treatment. More recently, we studied the expression of major histocompatibility complex (MHC) I and II, and the number and phenotypes of leukocytes in cerebral perivascular spaces (CPVS). We observed that natalizumab therapy was associated with a significant decrease in the cell surface expression of MHC class II molecules, and the numbers of dendritic cells in CPVS. In addition, no CD4(+) T cells were detectable in this compartment. Our observations may explain the differential and prolonged effects of natalizumab therapy on different leukocyte subsets in the central nervous system. They also suggest that natalizumab treatment may result in prolonged immunosuppression in peripheral organs, and the delayed onset of adverse events. PMID- 19300962 TI - Clinical trials in multiple sclerosis: current and future requirements - potential pitfalls. AB - Several lines of evidence support early immunomodulatory treatment of relapsing multiple sclerosis with either recombinant beta-interferons or glatiramer acetate and positive results from phase III trials encourage start of treatment even in patients with clinical isolated syndrome (CIS). However, currently available drugs for basic therapy are only partially effective and patients may still encounter relapses or disease progression. As treatment-refractory, clinically active MS can quickly lead to irreversible neurological disability there is an urgent need for more effective therapeutic strategies.The major goal is to reduce the damage, protect the tissue and enhance repair. In order to achieve these goals we not only need effective and safe drugs but also the appropriate study designs to really detect the most relevant outcomes [2, 7]. PMID- 19300963 TI - Isolated, relapsing and progressive demyelinating diseases of the central nervous system. AB - Many patients ask, "Can I get this again?" (a relapse) and "How bad is it? Will I end up in a wheelchair?" (severity/progression). These two questions guide this brief review of the clinical spectrum of demyelinating diseases of the central nervous system. PMID- 19300964 TI - Neuroprotection and neuroregeneration in multiple sclerosis. AB - Current treatments for multiple sclerosis (MS) include immunomodulatory or immunosuppressive agents but no neuroprotective or regenerative therapy is available. From various animal models, we have learned that remyelination in the CNS is a potent neuroprotective mechanism. The knowledge about oligodendrocyte biology and the process of remyelination has greatly increased in recent years; however, the precise mechanisms are far from being understood. Although remyelination is, in principle, also possible in the diseased MS brain, it is not clear why it fails in many MS patients. The clinical trials performed so far either failed to show an effect or were insufficient in design. Thus, further knowledge about the molecular mechanisms of the repair processes and MS pathophysiology is required to achieve the ultimate goal of a neuroprotective and neuroregenerative treatment in MS. PMID- 19300965 TI - Complementary and alternative medicine in multiple sclerosis. AB - Complementary and alternative medicine (CAM) is often used by patients with multiple sclerosis (MS) but has been disregarded in research until lately. Various confounding factors on CAM utilisation exist, though have hardly been investigated. Besides socio-demographic variables like education, income, gender and age, illness-related factors like severity of disease are discussed. Furthermore, the important role of psychological factors on CAM utilisation is described. In particular, coping strategies such as "rumination", "search for information" and "search for meaning in religion" are more often reported by CAM users than non-users and have important impact on CAM utilisation. PMID- 19300966 TI - Compliance, adherence, and the treatment of multiple sclerosis. AB - With the availability of disease-modifying, immunomodulatory therapies (DMT) for multiple sclerosis (MS) and the first long-term studies, it became obvious that problems of compliance to complex treatment regimens under chronic conditions would also apply to these approaches. In a selective overview, problems and findings of adherence research are depicted. Based on a discussion of basic concepts, issues of operationally defining and measuring adherence are outlined. Descriptive findings on adherence to DMTs and empirical predictors of nonadherence are then discussed. Referring to theoretical models of treatment motivation, selected problems (e. g., indication) and strategies of promoting adherence are described. Finally, implications of modern concepts of the patient therapist relationship for the issue of patient adherence are considered. PMID- 19300967 TI - Recent clinical trials and future therapies. AB - Immunotherapy for multiple sclerosis (MS) has developed extremely successfully during the past decade and a number of new strategies were developed for the treatment of the disease. Examples include therapeutic strategies targeting leukocyte differentiation molecules, costimulatory molecules, anti-adhesion molecules, chemotaxis, novel immunomodulators, autologous stem cell transplantation, anti-infectious therapies and strategies for neuroprotection, neurorepair and remyelination. Here we describe examples of monoclonal antibodies, a novel immunosuppressant and interesting neuroprotective strategies. PMID- 19300968 TI - What can we learn from failed clinical trials in multiple sclerosis? AB - The list of failed trials with ineffective or toxic drugs, despite encouraging data from research, is growing. Therapeutic trials in multiple sclerosis (MS) may be particularly vulnerable to fail because the pathogenesis of this disease is still not understood. Thus theoretically promising agents may paradoxically increase disease activity or be associated with unforeseen adverse effects. Short term favourable trends may reverse with a prolonged follow-up. Antigen-specific therapies can stimulate in vivo rather than to inhibit encephalitogenic cells. In addition to the negative message of failed trials itself, they can promote new hypotheses about disease pathophysiology. Failed trials can provide valuable information how studies and outcome measures should be designed for future trials. So it is important that negative trials are published and available for the MS community. PMID- 19300969 TI - National MS registries. AB - This review gives an overview of national registries that are currently in use for patients with multiple sclerosis (MS). The large-scale registries described herein include the Danish MS Registry, the Norwegian MS Registry, the Swedish MS Registry, the Italian MS Database Network, the North-American NARCOMS Registry, and the German MS Registry. These MS registries are extremely helpful for studying disease characteristics in large populations and monitoring the long term outcome of disease-modifying therapies. Furthermore, an almost complete ascertainment of cases provides information on the provision of treatments, services and supplies within a given area that may be used to compare different levels of health care within and between these regions. In the long-term, MS registries monitor the health care situation of MS patients over time including the implementation of guidelines relating to care and treatment, measure the improvements that have taken place, and reveal shortages and/or misalignment in health care services. The information gathered herein is not only useful for the long-term follow-up of the individual patient, but also for society as a whole by increasing understanding of and knowledge about MS and allowing national authorities and relevant parties to make informed and relevant decisions about MS. PMID- 19300970 TI - Cerebral MRI lesions and anti-tumor necrosis factor-alpha therapy. AB - We discuss two cases receiving different anti-tumornecrosis-factor alpha antagonists (anti-TNF-alpha); one for psoriatic arthritis (PA) and the other for ankylosing spondylitis (AS). Due to neurological symptoms cerebral magnetic resonance imaging (MRI) was performed and cerebral lesions were detected. Our interpretations of these cerebral lesions and the resulting diagnostic and therapeutic consequences are presented in regard of data published in the medical literature. PMID- 19300971 TI - Muscle rupture caused by exacerbated spasticity in a patient with multiple sclerosis. AB - We report a case of muscle rupture caused by acute exacerbation of spasticity in a patient with primary chronic progressive multiple sclerosis (PPMS). A complete disruption of the adductor muscles was diagnosed by sonography and still reproducible in a follow-up three months after the clinical event. To our knowledge, this is the first case report of muscle rupture caused by spasticity in a patient with MS. In addition to the clinical case report we give a short overview of morphological and functional changes in spastic muscle and current standards of symptomatic therapy. PMID- 19300972 TI - Paediatric multiple sclerosis: the experience of the German Centre for Multiple Sclerosis in Childhood and Adolescence. AB - Multiple sclerosis (MS) is a chronic inflammatory demyelinating disorder of the central nervous system and the most common disabling neurological disease in young adults. An estimated 5 % of MS patients already have first clinical symptoms before the age of 16 years (paediatric MS). In the paediatric age group comprehensive analysis of the natural clinical course and the course under treatment in a large MS cohort is still missing. We describe a cohort of paediatric MS patients treated in the German Centre for Multiple Sclerosis in Childhood and Adolescence. A total of 166 patients with definite MS who are registered in our database were analysed. The observation time was up to 14.9 years with a mean follow-up of 4.1 years. Median age was 12.4 years (range 4 to 18 years). Prior to puberty the gender ratio was almost equal, while in adolescence there was a strong female predominance as is seen in adult onset MS. Almost all patients presented with relapsing- remitting MS. The course of the disease was more benign than in adult MS with a very slow EDSS increase and complete remission after most relapses. Most patients received immunomodulative treatment with interferon-beta or glatiramer acetate and, in severe cases, natalizumab. However, adequate treatment guidelines for this age group are still lacking. PMID- 19300973 TI - Diagnostic algorithm for the differentiation of leukodystrophies in early MS. AB - Leukodystrophies are chronic progressive inherited white matter diseases frequently combined with an inborn error of metabolism. Some leukodystrophies clinically resemble chronic variants of multiple sclerosis (MS), while others exhibit multifocal MRI changes mimicking white matter changes known from MS imaging studies. The risk of misdiagnosing leukodystrophy as being MS is especially high in early disease stages comprising the possibility of initiating an inadequate therapy. Patients with clinical isolated syndromes should carefully be screened for positive family history, atypical clinical presentation or MRI pattern, symmetric involvement of long spinal tracts and peripheral nerves using evoked potentials and nerve conduction velocities and lack of oligoclonal bands in CSF all of which are "red flags" which consecutively may implicate consideration of leukodystrophy. Genetic or biochemical testing is widely available giving rise to a specific diagnosis in most patients. PMID- 19300974 TI - [Rosuvastatin in patients with chronic heart failure]. PMID- 19300975 TI - [Medication and falls in old age]. AB - Falls with and without injuries in elderly persons commonly have multiple causes. Exposure to drugs does contribute to these causes. Therefore, complete assessment and evaluation of prescription and over the counter drugs are essential parts of fall-prevention concepts. Frail elderly persons frequently treated with several medications are particularly predisposed to adverse drug effects which may increase the risk of falling. Risk increasing drug effects are dose dependent which have been best studied with psychotropic medication. Apart from psychotropic drugs, cardiovascular drugs contribute to FRIDs (Fall-Risk Increasing Drugs). Fall risk is particularly increased with drugs of the same therapeutic class combined or combinations of psychotropics and cardiovascular drugs. Intervention studies on withdrawal and dose reduction of fall-risk increasing drugs were successful in reducing the risk of falling. There is relatively few knowledge on whether and how drug treatment does decrease fall risk in elderly patients by improving safe mobility and walking ability relevant to activities of daily living. PMID- 19300976 TI - [Special features of pharmacotherapy in older hypertensive subjects]. AB - The prevalence of arterial hypertension in Germany is 60-80% in individuals between 65 and 74 years of age. In older people there is also a positive correlation between blood pressure level and rise in mortality due to myocardial infarction and stroke. Antihypertensive therapy can considerably improve the prognosis of older hypertensive individuals. Treatment is of enormous importance in these patients, particularly for prevention of strokes, which will continue to increase in the future as a result of demographic trends. Older patients have an elevated risk of developing undesirable drug reactions while under therapy such as a drop in blood pressure in an erect/standing position. Taking into account individual risk factors for unwanted events and avoidance of drug interactions during polypharmacotherapy are important aspects in differential therapy with antihypertensive agents in the elderly. PMID- 19300977 TI - [Free skin grafts as alternatives in reconstructive plastic surgery of the genitalia]. AB - PROBLEM: In the field of plastic/reconstructive urology, surgery often concerns the correction of congenital anomalies. Besides anomalies of the upper urinary tract these are disorders of the external genitalia like hypospadias and more seldom epispadias or a buried penis in males as well as complete vaginal aplasia or nondevelopment of the distal part of the vagina in different forms of disorders of sexual differentiation in females. The use of local tissue is the first choice for reconstruction. However, there is not always enough local tissue or it has already been used during unsuccessful interventions before. In these cases the use of a free skin graft should be considered. PATIENTS AND METHODS: We describe the cases of nine patients in whom a free skin graft was used for reconstruction. Two male patients had surgery because of a buried penis, and the third male patient underwent another revision after several unsuccessful corrections of epispadias. In two female patients with adrenogenital syndrome (AGS) construction of the distal vagina was necessary. Four patients had vaginal aplasia and needed a complete neovagina. In the male patients the free skin graft was harvested from the scrotum and in the female patients from the lower abdominal wall. RESULTS: In all nine patients the free skin graft is revascularized. The male patients show a good cosmetic result. The penile skin is mobile, which is also reflected by an unproblematic erection in the adult patient. The only complication in the female patients was a scar between the free skin graft and the skin of the proximal vagina in a patient with AGS. This scar was incised at 5 and 7 o'clock. CONCLUSION: A free skin graft offers the possibility of effectively covering areas when there is insufficient local tissue. PMID- 19300978 TI - [The value of real-time elastography in the diagnosis of prostate cancer]. AB - Randomized biopsy sampling under transrectal ultrasound (TRUS) guidance is the gold standard for the diagnosis of prostate cancer. In addition improvements in the quality of conventional ultrasound, new methods that complement conventional TRUS are opening the door to earlier and better targeted diagnosis of prostate cancer. One of these new methods is sonoelastography. Its impact on prostate cancer diagnostics has not yet been fully investigated, but the number of publications on this new technique indicate increasing interest in it. PMID- 19300979 TI - [Prognostic factors for survival of patients with metastatic renal cell carcinoma]. AB - Prognostic factors for the survival of patients undergoing immunotherapy for metastatic renal cell carcinoma (RCC) were first published in 1999 by Motzer et al. Since then several modifications of the prognostic model have been published. Even if not validated for targeted therapies stratification of RCC patients based on the prognostic factors provided the basis for recent clinical trials investigating the efficacy of sorafenib, sunitinib, temsirolimus, or bevacizumab/interferon. In order to compare the results of the trials and to generate treatment algorithms one has to carefully examine the prognostic factors used in the trials. The present article reviews the development of the prognostic models and their use in recent clinical trials. Furthermore, a reasonable algorithm for the treatment of patients with metastatic RCC is presented. PMID- 19300981 TI - Lithium treatment in Alzheimer's disease does not promote cognitive enhancement, but may exert long-term neuroprotective effects. PMID- 19300983 TI - Macrocyclic antibiotics as chiral selectors in the design of enantioselective, potentiometric membrane electrodes for the determination of S-flurbiprofen. AB - Construction of three novel enantioselective, potentiometric membrane electrodes based on carbon paste impregnated with different macrocyclic antibiotics vancomycin and teicoplanin as chiral selectors are described. The solutions for the construction of electrodes were prepared in phosphate buffer pH 4 for the vancomycin-based electrode (VCM), pH 6 and pH 6/40% acetonitrile solutions for teicoplanin-based electrodes, TCP I and II, respectively. The proposed electrodes were applied in the assay of S-flurbiprofen raw material and its pharmaceutical formulation by use of direct potentiometry, VCM electrode exhibiting the best enantioselectivity. The surfaces of the electrodes are easily renewable by simply polishing on an alumina paper. PMID- 19300982 TI - The future of endocannabinoid-oriented clinical research after CB1 antagonists. AB - INTRODUCTION: Great interest has been shown by the medical community and the public in the cannabinoid CB(1) receptor antagonists, such as rimonabant, for treatment of obesity, metabolic syndrome, and possibly drug addiction. DISCUSSION: This novel class of drug has therapeutic potential for other disorders, as the endocannabinoid system is involved in various health conditions. However, rimonabant, the first clinically available member of this class of drugs, has been linked to increased risk of anxiety, depression, and suicidality. Due to those risks, the European Medicines Agency called for its withdrawal from the market in October, 2008. Shortly after this decision, several pharmaceutical companies (Sanofi-aventis, Merck, Pfizer, Solvay) announced that they would stop further clinical research on this class of drug. Here, we provide an overview of those events and make several suggestions for continuing such clinical research, while safeguarding the safety of patients and clinical trial subjects. PMID- 19300984 TI - Recent advances in the development of novel materials for mycotoxin analysis. AB - For sensitive and specific toxin detection, mycotoxin immunoassays depend upon antibodies with high affinity and selectivity. While intact immunoglobulins remain the primary toxin-binding elements used in rapid assays, a number of alternatives have begun appearing in the literature. The alternatives can be broadly classified into those that are obtained by chemical synthesis and those that are obtained by altering biologically derived materials. Examples range from synthetically prepared polymers to recombinant fragments of antibodies, with a wide variety of synthetic and natural materials in-between. To date, obtaining the combination of selectivity and affinity needed for use in sensors has been more readily accomplished with biologically derived materials than with synthetic materials. Despite this, synthetic materials still offer certain potential advantages, such as high binding capacity and the ability to bind in environments that are too harsh for intact antibodies. This review focuses upon recent advances in the development of mycotoxin-binding materials and their potential for application in mycotoxin assays. PMID- 19300985 TI - Effects of seeing and hearing speech on speech production: a response time study. AB - Research demonstrates that listening to and viewing speech excites tongue and lip motor areas involved in speech production. This perceptual-motor relationship was investigated behaviourally by presenting video clips of a speaker producing vowel consonant-vowel syllables in three conditions: visual-only, audio-only, and audiovisual. Participants identified target letters that were flashed over the mouth during the video, either manually or verbally as quickly as possible. Verbal responses were fastest when the target matched the speech stimuli in all modality conditions, yet optimal facilitation was observed when participants were presented with visual-only stimuli. Critically, no such facilitation occurred when participants were asked to identify the target manually. Our findings support previous research suggesting a close relationship between speech perception and production by demonstrating that viewing speech can 'prime' our motor system for subsequent speech production. PMID- 19300986 TI - Deriving angular displacement from optic flow: a fMRI study. AB - Using fMRI we wished to identify brain areas subserving the conversion of velocity signals into estimates of self-displacement (velocity-to-displacement integration, VDI), a function which is a prerequisite for the ability to navigate without landmarks. As real self-motion is not feasible in an fMRI environment, we presented subjects with a ride along a circular path in virtual reality devoid of usable landmarks. We asked subjects to try and feel as if actually moving in the scene and to either detect and count changes in driving speed (V-task) or to estimate the angular displacement achieved during a ride (D-task). We examined the contrast between these two tasks with regard to two hypothesised key functions for VDI: (1) evoking an internal image of the self in space and (2) manipulating this image in proportion to perceived velocity at the pace of a time base. The BOLD-responses during both tasks were fairly similar showing activity with right hemispheric dominance in a large parieto-temporo-occipital area as well as in frontal and prefrontal areas. Contrast D-V revealed a mainly parieto hippocampal network comprising precuneus and inferior parietal cortex, posterior parieto-occipital cortex, retrosplenial cortex and the hippocampal region, but also right superior frontal gyrus and right cerebellum. It can be viewed as a blend of networks known to be involved in mental rotation and in navigation, except for the lack of ventral premotor and prefrontal activity. A tentative interpretation proposes a scenario where precuneus, together perhaps with posterior parieto-occipital cortex, provides the postulated mental image of the self in space and uses it to interpret results computed in the hippocampal region. In the hippocampal region, VDI proper would take place based on a map of spatial orientation, with the appropriate time scale being an intrinsic property. In addition, a dedicated time keeping system in inferior parietal cortex appears to be involved. PMID- 19300987 TI - Temporal dependence of in vivo USPIO-enhanced MRI signal changes in human carotid atheromatous plaques. AB - INTRODUCTION: Ultrasmall superparamagnetic iron oxide (USPIO)-enhanced MRI has been shown to be a useful modality to image activated macrophages in vivo, which are principally responsible for plaque inflammation. This study determined the optimum imaging time-window to detect maximal signal change post-USPIO infusion using T1-weighted (T1w), T2*-weighted (T2*w) and quantitative T2* (qT2*) imaging. METHODS: Six patients with an asymptomatic carotid stenosis underwent high resolution T1w, T2*w and qT2* MR imaging of their carotid arteries at 1.5 T. Imaging was performed before and at 24, 36, 48, 72 and 96 h after USPIO (Sinerem, Guerbet, France) infusion. Each slice showing atherosclerotic plaque was manually segmented into quadrants and signal changes in each quadrant were fitted to an exponential power function to model the optimum time for post-infusion imaging. RESULTS: The power function determining the mean time to convergence for all patients was 46, 41 and 39 h for the T1w, T2*w and qT2* sequences, respectively. When modelling each patient individually, 90% of the maximum signal intensity change was observed at 36 h for three, four and six patients on T1w, T2*w and qT2*, respectively. The rates of signal change decrease after this period but signal change was still evident up to 96 h. CONCLUSION: This study showed that a suitable imaging window for T1w, T2*w and qT2* signal changes post-USPIO infusion was between 36 and 48 h. Logistically, this would be convenient in bringing patients back for one post-contrast MRI, but validation is required in a larger cohort of patients. PMID- 19300988 TI - Vertebroplasty as treatment of aggressive and symptomatic vertebral hemangiomas: up to 4 years of follow-up. AB - INTRODUCTION: This study aimed to illustrate the validity of the treatment with vertebroplasty (VP) in patients with aggressive or symptomatic vertebral hemangioma (VH) with or without epidural extension. METHODS: From January 2003 to December 2007, 24 consecutive patients have been treated with VP, for a total of 36 vertebral bodies affected by VH: two cervical, ten dorsal, 24 lumbar. All the patients complained of a pain syndrome resistant to continuous medical medication; four of 24 patients also presented aggressive magnetic resonance features of the vertebral lesion and two patients showed also epidural extension. A unipedicular approach has been performed in 16 patients; a bipedicular approach has been performed in six, while for the cervical spine an anterior-lateral approach with manual dislocation of the carotid axis has always been performed. Bone biopsy was never done. All procedures have been carried out with local anesthesia, except for the treatment of the cervical hemangiomas which has always been performed under general anesthesia. Four vertebral bodies in the same session have been treated in one case. RESULTS: Results have been evaluated with the visual analog scale and the Oswestry Disability Index methods. In all the patients, in the following 24-72 h, a successful outcome has been observed with a complete resolution of pain symptom. Extravertebral vascular or discal cement leakage has been observed in four patients, without any onset of clinical radicular syndrome due to epidural diffusion. Clinical and radiological follow-up until 4 years has been performed in 12 patients and it showed stability of the treatment and absence of pain. CONCLUSIONS: Percutaneous treatment with VP for aggressive and symptomatic vertebral hemangiomas even with epidural extension is a valuable, mini-invasive, and quick method that allows a complete and enduring resolution of the painful vertebral symptoms without findings of fracture of a vertebral body adjacent or distant to the one treated. PMID- 19300989 TI - Calciuria, oxaluria and phosphaturia after ingestion of glucose, xylitol and sorbitol in two population groups with different stone-risk profiles. AB - The effects of glucose, sorbitol and xylitol ingestion on calciuria, oxaluria and phosphaturia in healthy black and white males on a standardized diet were investigated. After ingestion, they collected urine hourly for 3 h. Glucose decreased phosphaturia in blacks. Sorbitol decreased phosphaturia in both groups and increased oxaluria in whites. Xylitol increased oxaluria in blacks. Decreases in phosphaturia are attributed to penetration by phosphate into cells leading to decreases in phosphatemia and the renal filtered load. We suggest that this mechanism is more sensitive in blacks. We speculate that the increase in oxaluria after sorbitol ingestion occurs via its conversion to glyoxylate and that this pathway may be blocked in blacks. For the increase in oxaluria after xylitol ingestion, it is hypothesized that ketohexokinase and aldolase may be more active in blacks. Our results demonstrate, for the first time, a urinary effect due to sorbitol ingestion and an ethnic dependency of these and other effects. PMID- 19300990 TI - Custom-made covered transjugular intrahepatic portosystemic shunt (TIPS) in an infant with trisomy 22 and biliary atresia. AB - We report an 8-month-old girl with portal hypertension secondary to biliary atresia. The decision to treat with TIPS was made at the age of 8 months due to recurrent variceal bleeding. The procedure was carried out with a 6-mm bare stent due to her small size. Radiological follow-up with Doppler US showed gradual stenosis and finally occlusion of the stent 80 days after implantation. Revision was performed with placement of an additional 6-mm expanded polytetrafluoroethylene (e-PTFE) stent-graft that had remained patent for 9 months, proving that in small children with a portal vein diameter less than 8 mm, the combination of a bare stent and stent-graft can provide excellent results. PMID- 19300991 TI - Non-infective pulmonary disease in HIV-positive children. AB - It is estimated that over 90% of children infected with human immunodeficiency virus (HIV) live in the developing world and particularly in sub-Saharan Africa. Pulmonary disease is the most common clinical feature of acquired immunodeficiency syndrome (AIDS) in infants and children causing the most morbidity and mortality, and is the primary cause of death in 50% of cases. Children with lung disease are surviving progressively longer because of earlier diagnosis and antiretroviral treatment and, therefore, thoracic manifestations have continued to change and unexpected complications are being encountered. It has been reported that 33% of HIV-positive children have chronic changes on chest radiographs by the age of 4 years. Lymphocytic interstitial pneumonitis is common in the paediatric HIV population and is responsible for 30-40% of pulmonary disease. HIV-positive children also have a higher incidence of pulmonary malignancies, including lymphoma and pulmonary Kaposi sarcoma. Immune reconstitution inflammatory syndrome is seen after highly active antiretroviral treatment. Complications of pulmonary infections, aspiration and rarely interstitial pneumonitis are also seen. This review focuses on the imaging findings of non-infective chronic pulmonary disease. PMID- 19300993 TI - Chemically assisted microbial production of succinic acid by the yeast Yarrowia lipolytica grown on ethanol. AB - A new two-step process of production of succinic acid (SA) has been developed, which includes the microbial synthesis of alpha-ketoglutaric acid by the yeast Yarrowia lipolytica (step 1) and subsequent oxidation of the acid by hydrogen peroxide to SA (step 2). The maximum concentration of SA and its yield were found to be 63.4 g l(-1) and 58% of the ethanol consumed, respectively. The purity of the SA isolated from the culture liquid filtrate reached 100%. The yield of SA was as high as 82% of its amount in the culture liquid filtrate. The quality of the SA produced by the invented method meets the biochemical grade definitions, as is evident from the respiratory and other relevant parameters of rat liver mitochondria upon the oxidation of this SA. PMID- 19300994 TI - A periplasmic, pyridoxal-5'-phosphate-dependent amino acid racemase in Pseudomonas taetrolens. AB - The pyridoxal-5'-phosphate (PLP)-dependent amino acid racemases occur in almost every bacterium but may differ considerably with respect to substrate specificity. We here isolated the cloned broad substrate specificity racemase ArgR of Pseudomonas taetrolens from Escherichia coli by classical procedures. The racemase was biochemically characterized and amongst other aspects it was confirmed that it is mostly active with lysine, arginine and ornithine, but merely weakly active with alanine, whereas the alanine racemase of the same organism studied in comparison acts on alanine only. Unexpectedly, sequencing the amino-terminal end of ArgR revealed processing of the protein, with a signal peptide cleaved off. Subsequent localization studies demonstrated that in both P. taetrolens and E. coli ArgR activity was almost exclusively present in the periplasm, a feature so far unknown for any amino acid racemase. An ArgR derivative carrying a carboxy-terminal His-tag was made and this was demonstrated to localize even in an E. coli mutant devoid of the twin-arginine translocation (Tat) pathway in the periplasm. These data indicate that ArgR is synthesized as a prepeptide and translocated in a Tat-independent manner. We therefore propose that ArgR translocation depends on the Sec system and a post-translocational insertion of PLP occurs. As further experiments showed, ArgR is necessary for the catabolism of D: -arginine and D: -lysine by P. taetrolens. PMID- 19300995 TI - Potential of biofilm-based biofuel production. AB - Biofilm technology has been extensively applied to wastewater treatment, but its potential application in biofuel production has not been explored. Current technologies of converting lignocellulose materials to biofuel are hampered by costly processing steps in pretreatment, saccharification, and product recovery. Biofilms may have a potential to improve efficiency of these processes. Advantages of biofilms include concentration of cell-associated hydrolytic enzymes at the biofilm-substrate interface to increase reaction rates, a layered microbial structure in which multiple species may sequentially convert complex substrates and coferment hexose and pentose as hydrolysates diffuse outward, and the possibility of fungal-bacterial symbioses that allow simultaneous delignification and saccharification. More importantly, the confined microenvironment within a biofilm selectively rewards cells with better phenotypes conferred from intercellular gene or signal exchange, a process which is absent in suspended cultures. The immobilized property of biofilm, especially when affixed to a membrane, simplifies the separation of biofuel from its producer and promotes retention of biomass for continued reaction in the fermenter. Highly consolidated bioprocessing, including delignification, saccharification, fermentation, and separation in a single reactor, may be possible through the application of biofilm technology. To date, solid-state fermentation is the only biofuel process to which the advantages of biofilms have been applied, even though it has received limited attention and improvements. The transfer of biofilm technology from environmental engineering has the potential to spur great innovations in the optimization of biofuel production. PMID- 19300996 TI - Detoxification of model phenolic compounds in lignocellulosic hydrolysates with peroxidase for butanol production from Clostridium beijerinckii. AB - In the present study, we investigated the peroxidase-catalyzed detoxification of model phenolic compounds and evaluated the inhibitory effects of the detoxified solution on butanol production by Clostridium beijerinckii National Collection of Industrial and Marine Bacteria Ltd. 8052. The six phenolic compounds, p-coumaric acid, ferulic acid, 4-hydroxybenzoic acid, vanillic acid, syringaldehyde, and vanillin, were selected as model fermentation inhibitors generated during pretreatment and hydrolysis of lignocellulose. The enzyme reaction was optimized as a function of the reaction conditions of pH, peroxidase concentration, and hydrogen peroxide to substrate ratio. Most of the tested phenolics have a broad optimum pH range of 6.0 to 9. Removal efficiency increased with the molar ratio of H(2)O(2) to each compound up to 0.5-1.25. In the case of p-coumaric acid, ferulic acid, vanillic acid, and vanillin, the removal efficiency was almost 100% with only 0.01 microM of enzyme. The tested phenolic compounds (1 g/L) inhibited cell growth by 64-74%, while completely inhibiting the production of butanol. Although syringaldehyde and vanillin were less toxic on cell growth, the level of inhibition on the butanol production was quite different. The detoxified solution remarkably improved cell growth and surprisingly increased butanol production to the level of the control. Hence, our present study, using peroxidase for the removal of model phenolic compounds, could be applied towards the detoxification of lignocellulosic hydrolysates for butanol fermentation. PMID- 19300997 TI - Saturated norepinephrine transporter occupancy by atomoxetine relevant to clinical doses: a rhesus monkey study with (S,S)-[(18)F]FMeNER-D (2). AB - PURPOSE: In a previous PET study on norepinephrine transporter (NET) occupancy in the nonhuman primate brain, the relationship between NET occupancy and atomoxetine plasma concentration, and occupancies among different brain regions, were not demonstrated adequately. It may therefore be difficult to translate the results to the clinical situations. In the present study, the detailed change of NET occupancy was investigated among a wider range of doses in a more advanced manner. METHODS: Two rhesus monkeys were examined using a high-resolution PET system with (S,S)-[(18)F]FMeNER-D(2) under baseline conditions and after steady state infusion of different doses of atomoxetine (0.003 to 0.12 mg/kg per hour). NET occupancy of the thalamus, brainstem and anterior cingulate cortex was calculated using BP(ND) obtained with the simplified reference tissue model. RESULTS: NET occupancy increased regionally and uniformly as the plasma concentration of atomoxetine increased. The estimated Kd value (the amount to occupy 50% of NET) in the thalamus was 16 ng/ml. CONCLUSION: The results indicate that clinical doses of atomoxetine would occupy NET almost completely. PMID- 19300998 TI - Influence of multidrug resistance on (18)F-FCH cellular uptake in a glioblastoma model. AB - PURPOSE: Multidrug resistance, aggressiveness and accelerated choline metabolism are hallmarks of malignancy and have motivated the development of new PET tracers like (18)F-FCH, an analogue of choline. Our aim was to study the relationship of multidrug resistance of cultured glioma cell lines and (18)F-FCH tracer uptake. METHODS: We used an in vitro multidrug-resistant (MDR) glioma model composed of sensitive parental U87MG and derived resistant cells U87MG-CIS and U87MG-DOX. Aggressiveness, choline metabolism and transport were studied, particularly the expression of choline kinase (CK) and high-affinity choline transporter (CHT1). FCH transport studies were assessed in our glioblastoma model. RESULTS: As expected, the resistant cell lines express P-glycoprotein (Pgp), multidrug resistance-associated protein isoform 1 (MRP1) and elevated glutathione (GSH) content and are also more mobile and more invasive than the sensitive U87MG cells. Our results show an overexpression of CK and CHT1 in the resistant cell lines compared to the sensitive cell lines. We found an increased uptake of FCH (in % of uptake per 200,000 cells) in the resistant cells compared to the sensitive ones (U87MG: 0.89 +/- 0.14; U87MG-CIS: 1.27 +/- 0.18; U87MG-DOX: 1.33 +/- 0.13) in line with accelerated choline metabolism and aggressive phenotype. CONCLUSIONS: FCH uptake is not influenced by the two ATP-dependant efflux pumps: Pgp and MRP1. FCH would be an interesting probe for glioma imaging which would not be effluxed from the resistant cells by the classic MDR ABC transporters. Our results clearly show that FCH uptake reflects accelerated choline metabolism and is related to tumour aggressiveness and drug resistance. PMID- 19300999 TI - Chronic locked posterior shoulder dislocation with severe head involvement. AB - Arthroplasty is the treatment of choice for severely damaged humeral heads in chronic locked posterior dislocation of the shoulder. We retrospectively reviewed the results of 12 shoulder arthroplasties (11 patients) between 1999 and 2005. Mean follow-up was 37.4 +/- 16.8 months. Mean postoperative Constant score (CS) was 59.4 +/- 21.6 (normative age and gender-related CS 67.1 +/- 24). There was a significant improvement in range of motion for flexion, abduction and external rotation. There was negative correlation (Pearson's coefficient) between the related CS and number of previous operations, pain and duration of symptoms. One patient underwent revision surgery 36 months postoperatively with polyethylene insert exchange. There were two patients with mild and one with severe migration of the humeral head. Shoulder arthroplasty resulted in good midterm results for this patient group with benefits for range of motion, pain and patient satisfaction. PMID- 19301000 TI - Plating osteosynthesis of mid-distal humeral shaft fractures: minimally invasive versus conventional open reduction technique. AB - Results of two methods, conventional open reduction-internal plating and minimally invasive plating osteosynthesis (MIPO), in the treatment of mid-distal humeral shaft fractures were compared. Thirty-three patients were retrospectively analysed and divided into two groups. Group A (n = 17) patients were treated by MIPO and group B (n = 16) by conventional plating. The mean operation time in group A was 92.35 +/- 57.68 minutes and 103.12 +/- 31.08 minutes in group B (P = 0.513). Iatrogenic radial nerve palsy in group A was 0% (0/17) and 31.3% in group B (5/16 (P = 0.012). The mean fracture union time in group A was 15.29 +/- 4.01 weeks (range 8-24 weeks), and 21.25 +/- 13.67 weeks (range 10-58 weeks) in group B (P = 0.095). The mean UCLA end-result score in group A was 34.76 +/- 0.56 points (range 33-35), and 34.38 +/- 1.41 points (range 30-35) in group B (P = 0.299). The mean MEPI in group A was 99.41 +/- 2.43 points (range 90-100) and 99.69 +/- 1.25 points (range 95-100) in group B ( P = 0.687). When compared to the conventional plating techniques, MIPO offers advantages in terms of reduced incidence of iatrogenic radial nerve palsies and accelerated fracture union and a similar functional outcome with respect to shoulder and elbow function. PMID- 19301001 TI - Relationship of plasma and synovial fluid BMP-7 with disease severity in knee osteoarthritis patients: a pilot study. AB - The objective of this study was to investigate bone morphogenetic protein-7 (BMP 7) levels in both plasma and synovial fluid of patients with primary knee osteoarthritis (OA) and to determine their relationship to disease severity. Thirty-two patients with knee OA and 15 healthy subjects were enrolled in the study. Anteroposterior knee radiographs were taken to determine the disease severity of the affected knee. The radiographic grading of OA in the knee was performed using the Kellgren-Lawrence criteria. BMP-7 levels in the plasma and synovial fluid were measured using enzyme-linked immunosorbent assay. The mean plasma BMP-7 concentration of the knee OA patients was significantly higher compared with that of healthy controls (12.1 +/- 1.6 vs 3.5 +/- 0.9 pg/ml, P = 0.001). Although BMP-7 levels in plasma were higher with respect to paired synovial fluid samples, the difference was not statistically significant (12.1 +/ 1.6 vs 10.5 +/- 2.2 pg/ml, P = 0.3). Subsequent analysis showed that plasma BMP 7 levels significantly correlated with disease severity (r = 0.77, P < 0.001). Furthermore, the synovial fluid levels of BMP-7 also correlated with disease severity (r = 0.60, P < 0.001). In addition, plasma BMP-7 levels showed a positive correlation with synovial fluid BMP-7 levels (r = 0.71, P < 0.001). Overexpression of BMP-7 in plasma and synovial fluid is related to progressive joint damage in knee OA. These findings suggest that BMP-7 might serve as a biochemical parameter for determining disease severity in primary knee OA and could play a potential role in cartilage protection and repair of OA. PMID- 19301002 TI - Triple pelvic osteotomy in the treatment of Legg-Calve-Perthes disease. AB - This article presents the results of Legg-Calve-Perthes (LCP) disease treatment using triple pelvic osteotomy. Thirty patients were analysed. The conditions for inclusion in the study were complete medical documentation and follow-up until the disease was resolved. Postoperatively, no patients were immobilised. Rehabilitation was initiated early in all patients, and full weight bearing was allowed after ten weeks. With this method, an increase of the CE angle of 17.43 +/- 4.020 degrees was achieved. Containment was increased from an initial 6.67% to 53.33% of patients at the final check-up. Similar improvement was achieved by using Herring classification of the damage; preoperatively most hips belonged to group C, and postoperatively to group A. Postoperatively, functional results were also considerably improved. The authors recommend triple pelvic osteotomy as the method of choice in the treatment of severe cases of LCP disease. PMID- 19301003 TI - Effects of physical environment on the evolution of Kashin-Beck disease in Tibet. AB - In previous studies we observed a proximo-distal gradient of lesion frequencies along the limb, with the distal joints being the most often affected. This suggests an associated effect of environmental factors on the most exposed joints. On a population of 820 children (mean age 13 years) of endemic areas distributed in groups of healthy and severity stages I to III of KBD (Kashin-Beck disease), the effects of different working activities were studied. Heavy work like that of a ploughman were compared to light physical work, e.g. school children, and exposure to cold and history of frostbite were also considered. The most severe stages, II and III, were present in 72% of the ploughman vs. 29% of the schoolchildren, 70% of the shepherds vs. 30% (p < 0.001) of the schoolchildren, and in 65% of the shepherds working in winter vs. 40% of those working in the other seasons (p < 0.001). In the group with history of frostbite, 58% present the severest stages vs. 40% without (p < 0.001). The results confirm a highly significant relation between microtrauma and cold and the severity of the KBD alterations. PMID- 19301004 TI - Antimony-trioxide- and arsenic-trioxide-induced apoptosis in myelogenic and lymphatic cell lines, recruitment of caspases, and loss of mitochondrial membrane potential are enhanced by modulators of the cellular glutathione redox system. AB - During the last years remission rates of more than 72% for arsenic(III)-oxide (As(2)O(3)) treatment in relapsed or refractory acute promyelocytic leukemia have been published. As(2)O(3) is under clinical investigation for therapy of leukemia and solid tumors. Due to the chemical affinity of arsenic and antimony, we analyzed the potency of antimony(III)-oxide (Sb(2)O(3)) to exert As(2)O(3)-like effects. Based on the same molar concentrations, lower efficacy in apoptosis induction and caspase-independent decrease of mitochondrial membrane potential was observed for Sb(2)O(3). No difference in sensitivity to As(2)O(3) or Sb(2)O(3) was detected in CEM cells when compared to their multiple drug resistant derivatives. Apoptosis was induced by combining sub-apoptotic concentrations of Sb(2)O(3) or As(2)O(3) with sub-apoptotic concentrations of DL: -buthionine-[S,R]-sulfoximine (BSO). Other modulators of the cellular redox system showed this effect to a lower extent and enhancement was not consistent for the different cell lines tested. Caspase inhibitors protected cell lines from Sb(2)O(3)- and As(2)O(3)-induced apoptosis. When BSO was added, the inhibitors lost their protective ability. The ability of modulators of the cellular redox system in clinically applicable concentrations to enhance the apoptotic effects of the two oxides in a synergistic way may be helpful to reduce their toxicity by optimizing their dose. PMID- 19301005 TI - Outcome of HLA-matched related allogeneic hematopoietic stem cell transplantation for patients with acute leukemia in first complete remission treated in Eastern European centers. Better results in recent years. AB - The goal of this study was to analyze results and to determine factors affecting outcome of HLA-matched hematopoetic stem cells transplantation (MRD-HSCT) for patients with acute leukemia transplanted in first complete remission in Eastern European countries. Six hundred forty HSCT were performed between 1990 and 2006 for adults with acute myeloid (n = 459) and lymphoblastic (n = 181) leukemia. Two year leukemia-free survival (LFS), nonrelapse mortality (NRM), and relapse incidence were 58 +/- 2%, 19 +/- 2%, and 23 +/- 2%, respectively. The cumulative incidence of NRM decreased from 22 +/- 2% for patients treated between 1990 and 2002 to 15 +/- 3% for transplantations performed between 2003 and 2006 (p = 0.02), despite increasing recipient age. In a multivariate analysis, time of HSCT affected both NRM and LFS. Among other prognostic factors, the use of TBI decreased relapse incidence and increased the LFS rate. We conclude that results of MRD-HSCT for acute leukemia in Eastern Europe improved over time as a consequence of decreased NRM. The use of TBI containing regimens appears advantagous. PMID- 19301006 TI - Pre-clinical evaluation of Rh2 in PC-3 human xenograft model for prostate cancer in vivo: formulation, pharmacokinetics, biodistribution and efficacy. AB - PURPOSE: This study assesses the pharmacokinetics, biodistribution and efficacy of ginsenoside Rh2 as a single agent administered in a novel oral dosage formulation. METHODS: A novel oral dosage formulation of Rh2 has been described. Rh2 levels in blood and tissues following administration to nu/nu nude mice were determined by high performance liquid chromatography tandem mass spectroscopy. Efficacy was determined in an established PC-3 human prostate cancer model. RESULTS: Rh2 administered at a dose of 120 mg/kg exhibited a peak plasma concentration of 19.0 +/- 2.0 microg/ml. Rh2 levels were measurable in prostate and tumor tissues, with as much as 0.3% of the administered dose being detected in tumors. This formulation exhibited no measurable toxicity as judged by weight loss or changes in serum levels of aspartate aminotransferase, alanine aminotransferase, and creatinine. This dose engendered a significant delay in PC 3 tumor growth, an increase in apoptotic index, and a decrease in tumor cell proliferation. CONCLUSIONS: Rh2 is a stable compound that can be formulated for oral gavage. Pharmacokinetics studies demonstrate its ability to be absorbed following oral administration. Future studies will assess the pharmacokinetics of Rh2 when administered in combination with docetaxel. PMID- 19301007 TI - [Structure of amyloid fibrils]. AB - Amyloid fibrils are structurally defined as fibrillar polypeptide aggregates with a characteristic cross-beta structure. Such fibrils can be formed by certain polypeptide sequences in the human body and by numerous polypeptide sequences in vitro. All amyloid fibrils possess a structural spine that is formed by a cross beta structure. This structure is stabilized by hydrogen bonds between the polypeptide backbone. In recent years, various biophysical techniques, such as X ray crystallography, solid state nuclear magnetic resonance spectroscopy and electron cryo-microscopy have provided insights into the structural organization of amyloid fibrils. This review presents an overview of important results obtained with these methods. PMID- 19301008 TI - Whole-body MRI: a helpful diagnostic tool for juvenile dermatomyositis case report and review of the literature. AB - Juvenile Dermatomyositis (JDM) is a rare autoimmune disease in childhood. Distinction between muscle inflammation and a residual state can be difficult especially under immunosuppressive therapy and in active disease without correlating muscle enzyme tests or functional muscle scores. Our goal is to demonstrate the benefit of whole-body magnetic resonance imaging (MRI) as a diagnostic modality in the detection and management of JDM. One patient with JDM was monitored using clinical examination, muscle enzyme tests, muscle scores and whole-body MRI. During immunosuppression, the patient presented several times to our department without clear correlation between clinical picture, muscle enzyme tests and muscle scores. Whole-body MRI proved reliable in assessing the true state of the disease, thus providing invaluable information in the management of the inflammatory myopathy. This is of utmost importance for the therapeutic optimization in order to prevent further damage especially in children with active but subclinical disease. PMID- 19301009 TI - Systemic lupus erythematosus in Egyptian children. AB - The aim of the study was to study the characteristics of systemic lupus erythematosus (SLE) in the Egyptian population, comparing it to other populations. We retrospectively studied 207 patients with SLE diagnosed between 1990 and 2005. We obtained clinical features and laboratory data and analyzed them statistically. We studied 151 female and 56 male SLE patients. The female to male ratio was 2.7 to 1 and the mean age at presentation was 10 +/- 2.7 years (range 2-16). The mean disease duration was 6.47 +/- 3.74 years. At diagnosis, musculoskeletal, constitutional and mucocutaneous manifestations were the commonest features. During follow-up, the prevalence of nephritis (67%), hematological manifestations (44.9%), photosensitivity (44%), arthritis (39%), malar rash (38.2%), serositis (32.9%) and neuropsychiatric manifestations (24.25%) increased significantly. Those whose age of onset of the disease was 0.05). Neoplastic change was also not observed in these groups. Four groups were compared in terms of neoplastic effects and infections. In Groups 1, 5 rats were normal, two developed vulvovaginitis and endometritis with overinfection (in both ovaries), and one developed salpingitis (in both fallopian tubes), that is, infection was found in a total of two rats. In Group 2, only one experimental animal had endometritis. All the animals in Groups 3 and 4 developed infections. CONCLUSIONS: Talc has unfavorable effects on the female genital system. However, this effect is in the form of foreign body reaction and infection, rather than being neoplastic. PMID- 19301024 TI - A new metabolic scoring system for analyzing the risk of hypertensive disorders of pregnancy. AB - OBJECTIVE: The aim of this study was to investigate the relationship between some components of metabolic syndrome (MS) and pregnancy induced hypertension (PIH). STUDY DESIGN: Forty-one patients with PIH (gestational hypertension or preeclampsia) after 32 weeks of gestation were compared with 97 normotensive pregnant women. Metabolic scores (0-4) were created using standard deviations in normotensive cases: mean level + 1SD for BMI (>31 kg/m(2)), mean level + 1SD for triglyceride (>287 mg/dl), mean level + 1SD for fasting serum glucose (>90 mg/dl)) and mean level - 1SD for HDL (<48 mg/dl). RESULTS: The mean values for BMI (31.6 +/- 5.7 vs. 27.7 +/- 3.6; P < 0.0001), fasting triglyceride (341 +/- 129 vs. 220.7 +/- 67; P < 0.0001) and glucose (87.5 +/- 17.1 vs. 79.6 +/- 10.4; P = 0.0009) were higher in hypertensive group. The proportions of the women with a positive result for each of the components were significantly higher in the group of PIH. The percentage of the cases having 2 (35.2 vs. 8.2%; P = 0.0002) and 3 or more (27 vs. 4.1%; P = 0.0003) components of MS was higher in the hypertensive group and the percentage of the cases with none of these factors was high in the normotensive group (10.8 vs. 56.7%; P < 0.0001). CONCLUSION: The presence of multiple components of MS may be a risk factor in the development of PIH. New scoring systems according to the gestational age might be useful in analyzing the risk of PIH. PMID- 19301025 TI - Successful twin pregnancy in a unicornuate uterus with one fetus in the non communicating rudimentary horn. AB - Rudimentary horn is a rare anomaly and results from arrested development of the mullerian ducts. We report a case of twin pregnancy in a unicornuate uterus with a non-communicating rudimentary horn, in which two siblings each in separate horn were delivered successfully by caesarean section. PMID- 19301026 TI - Screening for gestational diabetes: usefulness of clinical risk factors. AB - OBJECTIVE: There is little consensus regarding selective or universal screening for gestational diabetes. The aim of this study is to determine the influence of risk factors on incidence of GDM in Iranian population by using 75 g OGTT. METHODS: At the first prenatal visit, 924 pregnant women were assessed for age, BMI, obstetric history, family history of diabetes, and diagnosis of polycystic ovary syndrome before pregnancy. All eligible women underwent 2-h 75 g oral glucose tolerance test. Gestational diabetes was diagnosed according to American Diabetes Association criteria. RESULTS: During study period, 68 [7.4% (95% CI 5.9 9.2)] cases of GDM were found. Age, pre-pregnancy BMI, and family history of DM were the independent clinical predictors of GDM. In women with age <25, BMI < or = 24.9, and negative family history for diabetes, the risk of GDM was 0.008 (0.001-0.044). This risk increased to 0.250 (0.102-0.495) in women with age > or =30, BMI > or = 30 and positive family history for diabetes. With selective screening and without screening in low risk group, we will do 13.6% (126 of 924) less screening tests while missing potentially 1.5% (1 of 68) of GDM cases. CONCLUSION: Age, BMI, and family history of diabetes were independent risk factors in developing gestational diabetes. Concerning these factors, we do not miss substantial number of GDM cases with selective screening. PMID- 19301027 TI - Post-thyroidectomy hemorrhage: a national study of patients treated at the Danish departments of ENT Head and Neck Surgery. AB - As bleeding in the neck region is a potentially life-threatening complication, we found it imperative to concretize the frequency and to identify possible reasons for this complication. A national database of all thyroid surgery performed inside the specialty of ENT Head and Neck Surgery (THYKIR) was established in January 2001. This nationwide cohort study represents 5,490 patients included until December 2007. Overall hemorrhage frequency was 4.2% with a wide variation among departments. Multiple regression analysis identified age, male gender, malignant histology and extent of surgery as independent risk factors for hemorrhage. Increased hospital stay and infection rates were found in patients treated with drainage. The median time for onset of postoperative hemorrhage was 3 h (range 0-105). Compared with international literature our incidence of post thyroidectomy hemorrhage is relatively high. Improvement might be reached by the exchange of experience between departments with focus on adequate surgical technique and careful hemostasis. PMID- 19301028 TI - Suppression of retinal neovascularization by the iNOS inhibitor aminoguanidine in mice of oxygen-induced retinopathy. AB - BACKGROUND: Retinal neovascularization (NV) is a major cause of blindness associated with ischemic retinal disorders. Our study was focused on evaluating the inhibitory effect of aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase (iNOS), on retinal NV in mice of oxygen-induced retinopathy (OIR). METHODS: An OIR model was established with 7-day-old C57BL/6J mice. One day before and 1 and 3 days after being returned to the room air, the right eyes were injected intravitreally with bevacizumab, AG or bevacizumab+AG respectively. The left eyes were injected with normal saline (NS) as control. The mice were killed at postnatal day 17 (P17). The effects of AG or bevacizumab on iNOS or VEGF expressions were evaluated by RT-PCR and immunohistochemistry. Retinal NV was examined by fluorescein angiography, and was quantified histologically by CD34 immnunostaining at P17. RESULTS: Compared with NS-treated eyes, retinal VEGF and iNOS mRNA expressions were significantly reduced in AG- and bevacizumab+AG treated eyes; whereas in bevacizumab-treated eyes, retinal VEGF mRNA expression increased and iNOS mRNA expression remained unchanged. The above changes were confirmed by immunohistochemical study. The generalized decrease in both VEGF and iNOS distributions in mice retina treated with AG or bevacizumab+AG was demonstrated by immunohistochemistry. Retinal NV was significantly reduced in all three groups treated with bevacizumab, AG or bevacizumab+AG, when compared with NS-treated eyes. CONCLUSIONS: iNOS activation plays a pathological role in retinal NV in a mouse model of ischemic retinopathy. Administration of AG significantly suppressed retinal NV. Therefore, AG appears to be a novel and effective therapeutic approach for retinal NV. PMID- 19301029 TI - Sedentary lifestyle as a risk factor for low back pain: a systematic review. AB - OBJECTIVES: To review systematically studies examining the association between sedentary lifestyle and low back pain (LBP) using a comprehensive definition of sedentary behaviour including prolonged sitting both at work and during leisure time. METHODS: Journal articles published between 1998 and 2006 were obtained by searching computerized bibliographical databases. Quality assessment of studies employing a cohort or case-control design was performed to assess the strength of the evidence. RESULTS: Using pre-determined keywords, we identified 1,778 titles of which 1,391 were considered irrelevant. Then, 20 of the remaining 387 publications were scrutinized for full review after an examination of all the 387 abstracts. Finally, 15 studies (10 prospective cohorts and 5 case-controls) were included in the methodological quality assessment, of which 8 (6 cohorts and 2 case-controls; 53%) were classified as high-quality studies. One high-quality cohort study reported a positive association, between LBP and sitting at work only; all other studies reported no significant associations. Hence, there was limited evidence to demonstrate that sedentary behaviour is a risk factor for developing LBP. CONCLUSIONS: The present review confirms that sedentary lifestyle by itself is not associated with LBP. PMID- 19301030 TI - Genetic analysis of functional redundancy of BRM ATPase and ATSWI3C subunits of Arabidopsis SWI/SNF chromatin remodelling complexes. AB - In yeast and mammals, ATP-dependent chromatin remodelling complexes of the SWI/SNF family play critical roles in the regulation of transcription, cell proliferation, differentiation and development. Homologues of conserved subunits of SWI/SNF-type complexes, including Snf2-type ATPases and SWI3-type proteins, participate in analogous processes in Arabidopsis. Recent studies indicate a remarkable similarity between phenotypic effects of mutations in the SWI3 homologue ATSWI3C and bromodomain-ATPase BRM genes. To verify the extent of functional similarity between BRM and ATSWI3C, we have constructed atswi3c brm double mutants and compared their phenotypic traits to those of simultaneously grown single atswi3c and brm mutants. In addition to inheritance of characteristic developmental abnormalities shared by atswi3c and brm mutants, some additive brm-specific traits were also observed in the atswi3c brm double mutants. Unlike atswi3c, the brm mutation results in the enhancement of abnormal carpel development and pollen abortion leading to complete male sterility. Despite the overall similarity of brm and atswi3c phenotypes, a critical requirement for BRM in the differentiation of reproductive organs suggests that its regulatory functions do not entirely overlap those of ATSWI3C. The detection of two different transcript isoforms indicates that BRM is regulated by alternative splicing that creates an in-frame premature translation stop codon in its SNF2-like ATPase coding domain. The analysis of Arabidopsis mutants in nonsense-mediated decay suggests an involvement of this pathway in the control of alternative BRM transcript level. PMID- 19301031 TI - Expression of prostatic acid phosphatase (PSAP) in transurethral resection specimens of the prostate is predictive of histopathologic tumor stage in subsequent radical prostatectomies. AB - Clinical management of incidental prostate cancer (IPC) remains challenging since its clinical course cannot be predicted by conventional histopathology. Aiming to define predictive factors in IPC, we correlated the immunohistochemically detected expression of prostate-specific antigen (PSA), prostatic acid phosphatase (PSAP), alpha-methylacyl-CoA racemase (AMACR, p504s), and androgen receptor in transurethral resection specimens with Gleason scores and histologic staging on the corresponding radicals in a cohort of 54 patients (mean age, 65.9 years; range, 49-80 years). PSAP expression showed a significant correlation with tumor staging (rho = -0.37; p = 0.02) but not with Gleason scores (rho = -0.06; p = 0.69). K-statistics revealed a highly significant moderate interobserver agreement concerning the evaluation of PSAP staining (K = 0.47; p < 0.001). In contrast, the other markers assessed failed to correlate with conventional histopathology. Therefore, PSAP might be predictive of tumor stage in IPC and represent a valuable adjunct for clinical decisions in terms of individual therapeutic management. PMID- 19301032 TI - Leiomyomatous nodules in a cellular angiofibroma: a hitherto unreported finding. AB - Cellular angiofibroma is a benign tumor of the superficial soft tissues of the vulvovaginal and inguinoscrotal regions of adult patients. Histologically, it is characterized by bland spindle cells arranged without any pattern in a stroma with thin collagen fibrils and evenly distributed small- to medium-sized blood vessels with hyalinized walls. The case of a 44-year-old female with a perineal cellular angiofibroma containing small leiomyomatous nodules is described in this case report. These nodules could probably be originated from smooth muscle differentiation of the spindle cell component of an otherwise conventional cellular angiofibroma. To our knowledge, the finding of distinct leiomyomatous nodules within a cellular angiofibroma has not been previously reported. PMID- 19301033 TI - Central European Vaccination Advisory Group (CEVAG) guidance statement on recommendations for the introduction of HPV vaccines. AB - Vaccines against human papillomavirus (HPV), the primary causative agent in cervical cancer, are licensed. This paper contains the Central European Vaccination Advisory Group (CEVAG) guidance statement on the introduction of HPV vaccines in central Europe. Eight countries currently have medical representatives on CEVAG: the Czech Republic, Estonia, Hungary, Lithuania, Poland, Romania, Slovakia and Turkey. By raising awareness and disseminating information, CEVAG aims to promote the efficient and safe use of vaccines to prevent, control and if possible eliminate infectious diseases. In January 2008, the European Centre for Disease Prevention and Control published a report entitled Guidance for the Introduction of HPV Vaccines in EU Countries. Members of CEVAG have taken the information relevant to their countries from this report and, with consideration of local issues, produced these guidance recommendations for the introduction of HPV vaccines in the CEVAG region, which may be adapted for use in individual countries. PMID- 19301034 TI - A prospective 7-year survey on central venous catheter-related complications at a single pediatric hospital. AB - The aims of this study were to assess the incidence and risk factors of major central venous catheter (CVC)-related complications in a large cohort of children affected by oncological, hematological, or immunological diseases in a 7-year prospective observational study at a single center. Nine hundred fifteen CVCs were inserted in 748 children for a total period of 307,846 CVC-days. Overall, 298 complications were documented with a complication rate of 0.97/1,000 CVC days: 105 mechanical complications (dislocations 0.30/1,000 CVC-days, ruptures 0.04/1,000 CVC-days), 174 infections (bloodstream infections 0.46/1,000 CVC-days, tunnel infections 0.10/1,000 CVC-days), and 19 thrombosis (0.06/1,000 CVC-days). Significant risk factors were: diagnosis of acute lymphoblastic leukemia (ALL) and age or =5 days after the onset of the symptoms of the disease (late use of ribavirin, LUR); and the last group of 11 patients did not receive ribavirin (no use of ribavirin, NUR). At 5-10 days from the onset of symptoms the mean platelet counts of the patients in the EUR group were significantly higher than those of the patients in LUR group, and at 7-9 days, they were significantly higher than that of the patients in the NUR group. The mean aspartate transferase levels in the EUR group were significantly lower than of the NUR group on days 8 and 9, and the mean alanine transferase level was significantly lower on day 8 after the onset of the symptoms. There is a beneficial effect of ribavirin if given at an early phase of the CCHF. We suggest ribavirin use especially in the early phase of the disease. PMID- 19301048 TI - Extremity CT angiography: application to trauma using 64-MDCT. AB - Evolving multi-row detector computed tomography (MDCT) technology has resulted in increasing utility of CT angiography (CTA) in extremity vascular trauma diagnosis and characterization. Given the widespread availability as well as the ease of acquiring CTA in the trauma setting, CTA is increasingly being used as the initial diagnostic evaluation in extremity vascular trauma, replacing digital subtraction angiography in many institutions. One of the significant advantages of the application of 64-MDCT to extremity vascular trauma is the ability to integrate CTA into routine trauma torso protocols. This ultimately yields an efficient, tailored examination to evaluate the multi-trauma patient in a timely manner, a critical concern in this patient population. Although 64-MDCT offers increasing improvement in image quality, technical quality hinges on strict attention to protocol considerations in its implementation. This review article aims to detail the myriad injuries, which may be detected and characterized with CTA. Protocol considerations in the application of 64-MDCT technology to extremity vascular trauma are discussed, drawing from our experience in an urban, Level I trauma center. Finally, the advantages and techniques of integrating extremity CTA into torso trauma protocols will be described. PMID- 19301049 TI - Stroop effects on redemption and semantic effects on confession: simultaneous automatic activation of embedded and carrier words. AB - The present study was aimed at assessing whether focusing attention on a task relevant part of a word prevents processing of its meaning. Participants performed a color-naming task on a prime word followed by lexical decision on a probe. Primes were words, which could contain an embedded color word (e.g., "redemption") written in an incongruent color. Probes were either semantically related (e.g., "confession") or unrelated (e.g., "production") to the prime word. A Stroop effect emerged for color words appearing either in the initial or in the final position of the carrier word. A priming effect also emerged, with faster responses to probes semantically related to the prime. These results are evidence that focusing attention on part of a prime (i.e., the embedded color word) does not prevent the semantic processing of the entire word. PMID- 19301050 TI - Changes in chromatic pattern-onset VEP with full-body inversion. AB - PURPOSE: Intra-ocular pressure (IOP) increases to double that of its normal level under full-body inversion, in part simulating high IOPs found in glaucoma and ocular hypertension. Inversion also simulates negative g-forces experienced in aerobatic maneuvers such as those produced during aerial combat. Studies using achromatic pattern-reversal visual evoked potentials (VEPs) have shown losses in response amplitude when subjects are inverted and IOP is increased. In other studies, chromatic, pattern-onset VEPs have been shown to be a sensitive and objective indicator of ocular and systemic pathology. Thus, chromatic pattern onset VEPs may also show changes in amplitude and/or latency when subjects are subjected to full-body inversion. METHODS: In this study we employed chromatic, pattern-onset VEPs to determine if there were changes in response latency for the different visual pathways with healthy subjects during full-body inversion. Stimuli were 1 cpd horizontal sine-wave patterns presented in an onset mode (100 ms on/400 ms off). Subjects (n = 7) were tested at low to medium contrast levels, which were subjectively equated across chromatic pathways (S, LM, and achromatic). Patterns were presented using LCD goggles. RESULTS: All subjects showed a small but statistically significant increase in latency in the large negative component (CII) for both L - M and S - (L + M) pathways during inversion. The achromatic pathway also showed a statistically significant increase in latency to the positive component (CI) during inversion. CONCLUSIONS: These results demonstrate that changes in ocular and/or systemic physiology during full-body inversion can result in increased latencies of chromatic and achromatic pattern-onset VEPs. PMID- 19301051 TI - Extracting sources from noisy abdominal phonograms: a single-channel blind source separation method. AB - In this work we highlight a methodology that extracts sources from noisy single channel abdominal phonograms. First, an appropriate matrix of delays is constructed. Next, multiple independent components are calculated using the FastICA algorithm. Then these components are projected back to the measurement space and classified for recovering the sources of interest. Single-channel phonograms obtained from three different subjects were analysed. Results show successful extraction of foetal heart sounds (FHS), maternal respiration/pulse wave, and line noise. It is important to point out the high performance of the method for extracting the former two as separate sources; especially due to the fact that pulse wave and FHS may overlap as maternal and foetal QRSs do in the abdominal ECG. The most outstanding factor is that this is achieved using a single-channel method. So, this approach extracts physiological sources from noisy abdominal phonograms, and we believe it will be useful for surveillance, not only for foetal well-being but also for maternal condition. PMID- 19301052 TI - Mathematical modeling of electrical activity of uterine muscle cells. AB - The uterine electrical activity is an efficient parameter to study the uterine contractility. In order to understand the ionic mechanisms responsible for its generation, we aimed at building a mathematical model of the uterine cell electrical activity based upon the physiological mechanisms. First, based on the voltage clamp experiments found in the literature, we focus on the principal ionic channels and their cognate currents involved in the generation of this electrical activity. Second, we provide the methodology of formulations of uterine ionic currents derived from a wide range of electrophysiological data. The model is validated step by step by comparing simulated voltage-clamp results with the experimental ones. The model reproduces successfully the generation of single spikes or trains of action potentials that fit with the experimental data. It allows analyzing ionic channels implications. Likewise, the calcium-dependent conductance influences significantly the cellular oscillatory behavior. PMID- 19301059 TI - Recent in vitro findings of negative inotropy of pantoprazole did not translate into clinically relevant effects on left ventricular function in healthy volunteers. AB - PURPOSE: Reports on cardiac problems with oral proton pump inhibitors have caused extensive safety reviews by the US Food and Drug Administration. We provide additional data on acute cardiac effects of an intravenous application. METHODS: Echocardiography was performed in 18 healthy volunteers after administration of a common high-dose regimen of pantoprazole (80 mg i.v. bolus followed by 8 mg/h for 1 h) or placebo. DESIGN: The design included a randomized, double-blind, placebo controlled cross-over trial. RESULTS: Ejection fraction (%, mean +/- SE) in the treatment group (placebo group) was 60.7 +/- 1.1 (61.2 +/- 1.7) at baseline, and 62.6 +/- 1.1 (62.1 +/- 1.9), 64.7 +/- 1.6 (63.5 +/- 1.3), 62.6 +/- 1.6 (61.0 +/- 1.6) and 63.0 +/- 1.4 (61.8 +/- 1.5) at 7.5, 15, 30 and 60 min after bolus application, respectively (p = n.s.). Similarly, no significant changes were found for cardiac output, cardiac index, blood pressure and heart rate. In contrast, gastric pH that was used as a treatment control was significantly increased 60 min after the application of pantoprazole as compared to baseline and to placebo. CONCLUSIONS: Pantoprazole as injection is safe in healthy subjects with respect to cardiac contractile function. However, in view of recent reports of negative inotropy of the drug, further studies in heart failure patients are required. PMID- 19301060 TI - Degos' disease: a rare condition simulating rheumatic diseases. AB - Dego's disease is an uncommon thrombo-occlusive vasculopathy that presented with skin rash and thrombotic complications affecting internal organs that may simulate rheumatic diseases and may be brought to the attention of rheumatologists. We present here a case of a middle-aged woman who presented with acute bowel infarction, persistent fever, elevated inflammatory markers and reversed albumin/globulin ratio suspicious of systemic vasculitis clinically. The diagnosis of Dego's disease was made from the classical skin lesions which were pink to brown papules with central depression and surrounding violaceous rim that were distributed over the trunk and extremities. Histology showed typical wedge shaped infarction in the affected organs with endothelial proliferation and occlusion by thrombus. Our patient was put on aspirin but suffered from recurrent bowel infarction 1.5 years later and eventually succumbed to septic complications. PMID- 19301061 TI - Tuberculosis after anti-TNF therapy. PMID- 19301062 TI - A score of risk factors associated with ischemic digital ulcers in patients affected by systemic sclerosis treated with iloprost. AB - A single series of patients affected by systemic sclerosis (SSc) and cyclically treated with iloprost was reviewed in order to evaluate the incidence of digital ulcers (DUs) and to compare the characteristics between the patients with and without this painful and disabling vascular complication. The record charts of 85 SSc patients were revised. Ischemic DUs and scleroderma contracture ulcers were separately considered. Twenty-nine subjects developed ischemic DUs during the course of the disease; whereas, scleroderma contracture ulcers occurred in six subjects. Ischemic DUs were associated with younger age at scleroderma onset, a longer disease duration, a longer time delay from scleroderma diagnosis to iloprost therapy, a bigger skin involvement, the presence of joint contractures, a videocapillaroscopic late pattern, a history of smoking, and of corticosteroids therapy. After the exclusion of four subjects with concomitant peripheral arterial disease, a forward-stepwise logistic regression analysis showed that only four variables, i.e., age at scleroderma onset, delay in beginning iloprost therapy, history of smoking, and presence of joint contractures remained significantly associated with ischemic DUs. In a score reflecting the sum of these four risk factors, the prevalence of ischemic DUs increased progressively from the lowest to the highest value of the score. The predictivity of this model was evaluated by the receiver-operating characteristics curve, with an estimated area under the curve of 0.836 with 95% confidence interval from 0.736 to 0.937. All the patients with scleroderma contracture ulcers were characterized by both diffuse pattern of disease and positivity for anti-Scl70 antibody. In this retrospective study, scleroderma patients with ischemic DUs are characterized by early disease onset, delay in beginning iloprost therapy, smoking habit, and presence of joint contraction. A score reflecting the sum of these factors may be useful to predict the risk of developing ischemic DUs. PMID- 19301063 TI - Heterozygous promotor haplotype LXA/LYB in MBL-deficiency associated with myopathy and left ventricular hypertrabeculation/noncompaction. AB - AIM: To report the genetic background of mannose-binding lectin (MBL)-deficiency in a patient with recurrent infections, cardiac disease, and myopathy. METHOD: Case report. RESULTS: In a 47-year-old male with recurrent respiratory infections, MBL-deficiency was diagnosed. He additionally had developed left bundle-branch-block, ventricular runs, and dilative cardiomyopathy. Left ventricular (LV)-hypertrabeculation and intra-myocardial calcifications were detected earlier. At age 44 years, unclassified myopathy, manifesting as easy fatigability, myalgias, and ptosis was diagnosed. After death from a sepsis with Staphylococcus aureus, autopsy revealed endocardial fibrosis and calcification, located over the compacted as well as non-compacted segments. The patient carried the heterozygous haplotype LXA/LYB in the MBL gene. MBL-deficiency was considered responsible for recurrent pulmonary infections and sepsis. The association between MBL-deficiency, LV-hypertrabeculation, endocardial fibrosis, and calcification remains speculative. CONCLUSIONS: MBL-deficiency due to the LXA/LYB genotype may be associated with recurrent pulmonary infections and fatal sepsis. Endocardial fibrosis and calcification results rather from LV-hypertrabeculation than MBL-deficiency. PMID- 19301064 TI - Abstracts of the Institute of Obstetricians & Gynaecologists Four Provinces Meeting and the Journal Obstetric & Gynaecology Society Annual Scientific Meeting. November 28, 2008. Dublin, Ireland. PMID- 19301065 TI - Assessment of water quality using chemometric tools: a case study of river Cooum, South India. AB - Multivariate statistical techniques were applied to identify and assess the quality of river water. Thirty samples were collected from the River Cooum, and basic chemical parameters--such as pH, effect concentration, total dissolved solids, major cations, anions, nutrients, and trace metals--were evaluated. To evaluate chemical variation and seasonal effect on the variables, analysis of variance and box-and-whisker plots were performed. Cluster analysis was applied, and pre-monsoon and post-monsoon major and minor clusters were classified. The relations among the stations were highlighted by cluster analysis, which were represented by dendograms to categorize different levels of contamination. Cluster analysis clearly grouped stations into polluted and unpolluted regions. The analysis classified the upper part of the river course into one unpolluted cluster; the middle and lower parts of the river clustered together, reflecting the presence of pollution. Factor analysis revealed that water quality is strongly affected by anthropogenic activities, rock-water interaction, and saline water intrusion. Seasonal variations in water chemistry were clearly highlighted by both cluster and factor analysis. Factor-score diagrams were used successfully to delineate the stations under study by the contributing factors, and seasonal effects on the sample stations were identified and evaluated. These statistical approaches and results yielded useful information about water quality and can lead to better water resource management. PMID- 19301066 TI - Diversity of both the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China sea. AB - Protease-producing bacteria are known to play an important role in degrading sedimentary particular organic nitrogen, and yet, their diversity and extracellular proteases remain largely unknown. In this paper, the diversity of the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea was investigated. The richness of the cultivable protease-producing bacteria reached 10(6) cells/g in all sediment samples. Analysis of the 16S rRNA gene sequences revealed that the predominant cultivated protease-producing bacteria are Gammaproteobacteria affiliated with the genera Pseudoalteromonas, Alteromonas, Marinobacter, Idiomarina, Halomonas, Vibrio, Shewanella, Pseudomonas, and Rheinheimera, with Alteromonas (34.6%) and Pseudoalteromonas (28.2%) as the predominant groups. Inhibitor analysis showed that nearly all the extracellular proteases from the bacteria are serine proteases or metalloproteases. Moreover, these proteases have different hydrolytic ability to different proteins, reflecting they may belong to different kinds of serine proteases or metalloproteases. To our knowledge, this study represents the first report of the diversity of bacterial proteases in deep-sea sediments. PMID- 19301067 TI - Don't fight the site: three geomorphic considerations in catchment-scale river rehabilitation planning. AB - Three geomorphic considerations that underpin the design and implementation of realistic and strategic river conservation and rehabilitation programs that work with the nature are outlined. First, the importance of appreciating the inherent diversity of river forms and processes is discussed. Second, river dynamics are appraised, framing the contemporary behavioral regime of a reach in relation to system evolution to explain changes to river character and behavior over time. Third, the trajectory of a reach is framed in relation to downstream patterns of river types, analyzing landscape connectivity at the catchment scale to interpret geomorphic river recovery potential. The application of these principles is demonstrated using extensive catchment-scale analyses of geomorphic river responses to human disturbance in the Bega and Upper Hunter catchments in southeastern Australia. Differing implications for reach- and catchment-scale rehabilitation planning prompt the imperative that management practices work with nature rather than strive to 'fight the site.' PMID- 19301069 TI - Tattoo: a navigational tool, a lighthouse for biomolecular level precision in surgical endoscopy. Newton's apple or Eve's? PMID- 19301068 TI - Pulmonary function and airway hyperresponsiveness in adults with sickle cell disease. AB - STUDY OBJECTIVES: Pulmonary involvement is a major cause of morbidity and mortality in patients with sickle cell disease (SCD). Although a high prevalence of airway hyperresponsiveness (AHR) has been reported, there are no studies demonstrating the relationship between AHR and acute chest syndrome (ACS) in adults with SCD. We investigated AHR prevalence, lung function abnormalities, and the relationships of these variables with ACS in SCD patients. METHOD: Thirty-one adult patients without asthmatic symptoms were compared with 31 matched controls. Expiratory flow rates, lung volumes, carbon monoxide diffusion capacity (DLCO), and methacholine provocation test (MPT) results were assessed. RESULTS: Forced vital capacity (FVC), forced expiratory volume in one second, forced expiratory flow rate at 25% to 75% of FVC (FEF(25%-75%)), peak expiratory flow rate, total lung capacity, and DLCO values were significantly lower in the patient group than in the controls. No significant difference in pulmonary function test results was found between patients with and without a history of ACS. Fifteen patients with SCD (48%) and only 5 controls (16%) had AHR (p = 0.007). A significant correlation was found between the number of ACS episodes and MPT positivity (r = 0.379, p = 0.035). The FEF(25%-75%) values were significantly lower in patients with positive MPT results than in patients with negative MPT results (p = 0.027). CONCLUSION: The prevalence of AHR was high in adult patients with SCD. A significant correlation was found between AHR and recurrent ACS episodes. Anti inflammatory controller agents can be used routinely to decrease pulmonary morbidity associated with SCD, even in the absence of asthmatic symptoms. PMID- 19301070 TI - Use of self-expandable metal stents for the treatment of esophageal perforations and anastomotic leaks. AB - BACKGROUND: Esophageal perforations and extensive anastomotic leaks after esophageal resection or gastrectomy are surgical emergencies with high mortality rates. In recent years, the use of self-expanding metallic stents (SEMS) has emerged as a promising treatment alternative for bridging and sealing the damage. This study aimed to evaluate the role of covered SEMS for the management of esophageal perforations and anastomotic leaks. METHODS: All esophageal stent placement procedures (174 procedures for 157 patients) at the authors' unit between January 1999 and April 2008 were assessed by a retrospective chart review. Of the 157 patients, 10 (6.4%) were treated with SEMS for sealing of an iatrogenic esophageal perforation (n = 4), a spontaneous esophageal rupture in Boerhaave's syndrome (n = 4), or an anastomotic leakage (n = 2). RESULTS: The median time from perforation or anastomotic leak to stent insertion was 13 days (range, 2 h to 48 days). The esophageal leak was totally sealed for 8 (80%) of 10 patients. The overall mortality rate was 50% (n = 5), and three (30%) of the five deaths were related to the perforation (n = 2) or leakage (n = 1). In both of the perforation cases, the diagnosis and treatment were substantially delayed. One patient with an anastomotic leak after gastrectomy died of the complication despite successful operative and SEMS treatment. Two of the deaths were unrelated to the perforation. In both cases, the cause of death was a disseminated malignant disease. CONCLUSIONS: Traumatic perforations and anastomotic leaks can be treated effectively with covered SEMS together with adequate drainage of the thoracic cavity even in cases of severely ill patients with inveterate esophageal perforations and leaks. PMID- 19301071 TI - Use of routine intraoperative endoscopy in elective laparoscopic colorectal surgery: can it further avoid anastomotic failure? AB - BACKGROUND: Anastomotic complications such as leakage and bleeding remain among the most serious complications of laparoscopic colorectal surgery. No perfect method exists for accurate and reliable avoidance of these catastrophes. This study aimed to study the usefulness of routine intraoperative endoscopy (RIOE) by comparing the surgical outcomes for RIOE patients with those for selective intraoperative endoscopy (SIOE) patients. METHODS: A retrospective chart review was performed for consecutive patients who underwent elective laparoscopic colorectal resections with distal anastomosis between January 2004 and May 2007. One surgeon performed RIOE, whereas the other three surgeons performed SIOE as necessary. All the abnormalities of IOE patients were managed with a subsequent salvage procedure, and the postoperative outcomes were compared between the RIOE and SIOE groups. RESULTS: The study included 107 patients in the RIOE group and 137 patients in the SIOE group. Abnormalities were detected in 11 RIOE patients (10.3%) (six with staple line bleeding, three with positive air leak test results, and two with additional pathology identified). All but one abnormality was laparoscopically managed without conversion to laparotomy. Whereas one patient experienced postoperative staple line bleeding that required a second operation, the remaining 10 patients recovered uneventfully. The mean hospital stay was 6 days (range, 4-9) days. The RIOE group had overall rates of 0% for anastomotic leakage and 0.9% for staple line bleeding. Intraoperative endoscopies were performed for 30 (21.9%) of the 137 patients in the SIOE group. The postoperative outcomes comparison between the RIOE and SIOE groups showed a tendency toward more overall anastomotic complications (0.9% vs. 5.1%) in the SIOE group, which due to the small sample size did not translate into significant differences in terms of staple line bleeding and anastomotic leakage. There also were no significant differences in other outcomes such as ileus, abdominal or pelvic sepsis, reoperation, positive distal margin, distance from distal margins, length of hospital stay, or mortality. CONCLUSIONS: Routine IOE for patients undergoing elective laparoscopic colorectal surgery with distal anastomosis can detect abnormalities at or around the anastomosis. Although the RIOE group had fewer postoperative anastomotic complications, due to the small sample size, the 5.7-fold increase in anastomotic failure did not translate into significantly better postoperative outcomes than the SIOE group experienced. A larger-scale single or multicenter prospective randomized study or a metaanalysis including similar studies is necessary for further investigation of this issue. PMID- 19301072 TI - Society of American Gastrointestinal Endoscopic Surgeons (SAGES) guidelines for office endoscopic services. PMID- 19301073 TI - Informed consent-'da Vinci code' for our safety in empowered patient's safety. PMID- 19301074 TI - Management of leak in the bariatric gastric bypass patient: reoperate, drain and feed distally. PMID- 19301075 TI - National trends and outcomes for the surgical therapy of ileocolonic Crohn's disease: a population-based analysis of laparoscopic vs. open approaches. AB - PURPOSE: The laparoscopic approach to Crohn's disease has demonstrated benefits in several small series. We sought to examine its use and outcomes on a national level. METHODS: All admissions with a diagnosis of Crohn's disease requiring bowel resection were selected from the 2000-2004 Nationwide Inpatient Sample. Regression analyses were used to compare outcome measures and identify independent predictors of undergoing laparoscopy. RESULTS: Of 396,911 patients admitted for Crohn's disease, 49,609 (12%) required surgical treatment. They were predominately Caucasian (64%), female (54%), and with ileocolic disease (72%). Most had private insurance (71%) and had surgery in urban hospitals (91%). Laparoscopic resection was performed in 2,826 cases (6%) and was associated with lower complications (8% vs. 16%), shorter length of stay (6 vs. 9 days), lower charges ($27,575 vs. $38,713), and mortality (0.2% vs. 0.9%, all P < 0.01). Open surgery was used more often for fistulas (8% vs. 1%) and when ostomies were required (12% vs. 7%). Independent predictors of laparoscopic resection were age <35 [odds ratio (OR) = 2.4], female gender (OR = 1.4), admission to a teaching hospital (OR = 1.2), ileocecal location (OR = 1.5), and lower disease stage (OR = 1.1, all P < 0.05). Ethnic category, insurance status, and type of admission (elective vs. non-elective) were not associated with operative method (P > 0.05). CONCLUSIONS: A variety of patient- and system-related factors influence the utilization of laparoscopy in Crohn's disease. Laparoscopic resection is associated with excellent short-term outcomes compared to open surgery. PMID- 19301076 TI - TGF-beta1 influence on TNF-alpha production and sTNF-Rs shedding in a coculture of colon carcinoma cell spheroids with normal cells. AB - In this study, the levels of TNF-alpha and its soluble receptors sTNF-Rp55 and sTNF-Rp75 were analyzed in cocultures of human colon carcinoma cell spheroids prepared from different grades of tumors with normal human colon epithelium, myofibroblast, and endothelial cell monolayers. Additionally, the influence of exogenously added rhTGF-beta1 (2 ng/ml) on the TNF-alpha and sTNF-Rs levels was tested. Direct interactions of colon carcinoma spheroids with normal cells caused decreases in TNF-alpha levels and normal cell-dependent changes in sTNF-Rs amounts as compared to normal cells cultured alone. The addition of rhTGF-beta1 to the cocultures caused a significant increase in TNF-alpha levels with a simultaneous decrease in the amounts of both sTNF-Rs. During direct interactions of colon carcinoma cells with normal tissue, paracrine effects are very important. We showed that TGF-beta1 acts synergistically with TNF-alpha and significantly limits sTNF-Rs shedding. Therefore, TNF-Rs bound to cellular membranes, but not their soluble forms, play an important role in tumor/normal cell interactions. TGF-beta1 and sTNF-Rs, in turn, may be valuable factors in colon cancer development and metastasis. PMID- 19301077 TI - A hybrid endo-laparoscopic therapy for common bile duct stenosis of a choledocho duodenostomy after a Roux-en-Y gastric bypass. AB - The essential growth of the number of Roux-en-Y gastric bypass procedures will obviously be accompanied by an increase of cases of common bile duct lithiasis. It seems evident that a close cooperation between surgeon and endoscopist will be needed on a routine basis. A laparoscopic-assisted transgastric ERCP is a well documented approach to investigate the pancreatico-biliary tree in patients where the duodenum has been bypassed as in Roux-en-Y gastric bypass. In this case we present the possibility of assisting the endoscopist not only by providing access to the gastric remnant but also by helping with laparoscopic instruments during duodenoscopy. A formally obese woman who had benefited from a RYGB developed recurrent jaundice despite a precedent common bile duct exploration and choledocho-duodenostomy. A laparoscopic-assisted transgastric endoscopy revealed an obstructed choledocho-duodenostomy caused by accretions around a migrated clip. The obstructing clip could be extracted by laparoscopic instruments under endoscopic control. PMID- 19301078 TI - Lipoprotein lipase expression in livers of morbidly obese patients could be responsible for liver steatosis. AB - BACKGROUND: Most patients with morbid obesity develop non-alcoholic fatty liver disease (NAFLD). The origins of lipid deposition in the liver and the effects of bariatric surgery in the obese with NAFLD are controversial. METHODS: We analyzed lipids and lipoprotein lipase (LPL) in both plasma and liver biopsies performed before and 12-18 months after Roux-en-Y gastric bypass surgery in 26 patients. RESULTS: In the livers of morbidly obese patients, the levels of LPL messenger RNA (mRNA) were higher (4.5-fold) before surgery than afterwards than control livers. In these patients, LPL activity was also significantly higher (91 +/- 7 mU/g) than in controls (51 +/- 3 mU/g, p = 0.0026) and correlated with the severity of the liver damage. All hepatic lipids were significantly increased in obese patients; however, after bariatric surgery, these lipids, with the exception of NEFA, tended to recover to normal levels. CONCLUSIONS: The liver of obese patients presented higher LPL activity than controls, and unlike the controls, this enzyme could be synthesized in the liver because it also present LPL mRNA. The presence of the LPL activity could enable the liver to capture circulating triacylglycerides, thus favoring the typical steatosis observed in these patients. PMID- 19301079 TI - Silicone-ring Roux-en-Y gastric bypass in the treatment of obesity: effects of laparoscopic versus laparotomic surgery on respiration. AB - BACKGROUND: The objective of this study was to compare the effects of silicone ring Roux-en-Y gastric bypass carried out by laparoscopy versus that accomplished by laparotomy on pulmonary function. METHODS: A total of 26 women (body mass index (BMI) 35-49 kg/m(2)) were studied candidates for silicone-ring Roux-en-Y gastric bypass carried out by laparoscopy (LG; n = 13) and laparotomy (or open surgery (OG); n = 13). Smokers, patients having lung disease, and those unable to carry out the tests properly were excluded. The physical therapy was standardized for both the groups. Respiratory evaluation was carried out during the preoperative period and on the second postoperative day by using spirometry and other tests that evaluated respiratory muscle strength and diaphragmatic mobility. Pain was evaluated by the visual analog scale on the second postoperative day. The statistical analysis was carried out with parametric or nonparametric tests, depending on the distribution of variables, considering p < 0.05 as statistically significant. RESULTS: Patients were similar with respect to age, BMI, and waist-to-hip ratio. A decrease in all variables was observed for both the groups in the postoperative period, although this decrease was less pronounced in the LG group. Pain intensity was also lower in the LG group. The length of hospital stay was 2 days, and there were no pulmonary complications. CONCLUSION: As there were no differences in the incidence of pulmonary complications and the length of hospital stay between the groups, the results showed that silicone-ring Roux-en-Y gastric bypass carried out by laparoscopy caused less pain and impairment of pulmonary function in the postoperative period. PMID- 19301080 TI - Case report: locked pubic symphysis: an open reduction technique. AB - A locked pubic symphysis occasionally occurs after a lateral compression injury of the pelvic ring. One pubic bone becomes entrapped behind the contralateral pubis. Lateral compression pelvic injuries are well recognized, but a lateral compression pelvic injury resulting in a locked pubic symphysis is rare. We describe a locked pubic symphysis with greater than 4 cm overlap that was reduced with simple maneuvers and readily available instruments. PMID- 19301081 TI - THA with a minimally invasive technique, multi-modal anesthesia, and home rehabilitation: factors associated with early discharge? AB - Multimodal anesthetic and pain regimens with minimally invasive surgical approaches and rapid rehabilitation protocols are thought to decrease length of stay after hip replacement. We asked whether a program including these three elements could achieve 23-hour discharge in a group of 665 patients and whether the length of hospital stay was influenced by patient age, gender, body mass index, change in hemoglobin or estimated blood loss, duration of surgery (< or = 90 or > 90 minutes), or American Society of Anesthesiologists physical status classification. Of the 665 patients, 259 (38.9%) were discharged home with indwelling peripheral nerve catheters. Hospital discharge in less than 24 hours was achieved in 295 (44.4%) of the 665 patients. After discharge, 73.5% of patients required no home or outpatient nursing care or physical therapy. Eighteen (2.7%) dislocations, eight (1.2%) femoral fractures requiring surgery, and thirteen (2.0%) revision procedures occurred within 90 days. Female gender, increasing age, increasing estimated blood loss, and American Association of Anesthesiologists classification 3 or 4 increased length of stay. Additional study is needed to confirm these factors and develop prospective prediction rules to allow for an outpatient approach to joint arthroplasty. LEVEL OF EVIDENCE: Level II, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19301082 TI - OP-1 augments glucocorticoid-inhibited fracture healing in a rat fracture model. AB - Glucocorticoids inhibit bone remodeling and fracture healing. We sought to determine whether osteogenic protein 1 (OP-1) can overcome this inhibition in a closed fracture model in the rat. Time-released prednisolone or placebo pellets were implanted subcutaneously; closed femoral fractures were created 2 weeks later in rats. Fractures received sham, OP-1 and collagen, or collagen-only implants. Femurs were harvested at 3, 10, 21, 28, and 42 days postfracture. Fractures were examined radiographically for amount of hard callus; mechanically for torque and stiffness (also expressed as a percentage of the contralateral intact femur); and histomorphometrically for amount of cartilaginous and noncartilaginous soft callus, hard callus, and total callus. Glucocorticoid administration inhibited fracture healing. The application of a devitalized Type I collagen matrix mitigated the inhibitory effects of prednisolone on fracture healing However, further increases in indices of fracture healing were observed when OP-1 was added to the collagen matrix compared with collagen alone. OP-1 and collagen was more effective than collagen alone. PMID- 19301083 TI - Haematoma in a hydrocele of the canal of Nuck mimicking a Richter's hernia. AB - We report a haematoma in a hydrocele of the canal of Nuck in a 69-year-old female. She presented with a right-sided groin swelling, the differential for which included an irreducible inguinal hernia or haematoma given her aspirin and clopidegrel use. Successful treatment involved evacuation of the haematoma with excision of the sac. Despite a high index of suspicion for a haematoma, these swellings should ideally be explored given the potential for co-existence of a hernia. PMID- 19301084 TI - Some aspects of the epidemiology of external hernias in Kumasi, Ghana. AB - BACKGROUND: In our communities there are large numbers of longstanding external hernias that remain untreated. This paper describes the epidemiological characteristics of these hernias. The data is expected to provide guidelines for sustained national and international efforts to reduce the burden of hernia by performing large-scale elective hernia repairs. METHOD: Between January 1998 and December 2007, a simple pro-forma was designed and used to record, in a prospective manner, the age, sex of patient and anatomical site of all external hernias seen and operated on both as emergencies and non-emergencies. These were patients who presented to a single general and paediatric surgeon at the Komfo Anokye Teaching Hospital in Kumasi, Ghana. RESULTS: A total of 2,506 patients were studied, of which 1,930 were male and 576 female, giving a male:female ratio of 3.4:1. Inguinal hernia was seen in 1,766 patients: 1,613 males and 153 females, a male:female ratio of 10.5:1. Children 4 years old or younger accounted for 20.9% of inguinal hernias. Femoral hernia was seen in 79 patients: 70 females and 9 males. These groin hernias were diagnosed in 1,845 patients, accounting for 73.6% of all patients. Incisional hernia was diagnosed in 380 patients (15.2%): 179 males and 201 females-a male:female ratio of 1:1.1. These two hernia types (groin and incisional) were seen in 2,225 patients, representing 88.8% of all the patients studied. All other hernias studied, including para-umbilical, umbilical and epigastric, were seen in 281 patients, representing 11.2% of the hernias studied. CONCLUSION: The epidemiology of external hernias seen and treated in our hospital is no different from that of hernias in other communities. Sustained efforts at elective repair will reduce the vast numbers of untreated accumulated hernias in our communities and thus prevent unnecessary morbidity and mortality. PMID- 19301085 TI - Preperitoneal memory-ring patch for inguinal hernia. Re: Preperitoneal memory ring patch for inguinal hernia: a prospective multicentric feasibility study, Berrevoet et al. (2009) Hernia (in press) doi: 10.1007/s10029-009-0475-4. PMID- 19301086 TI - A prospective, randomized, comparative trial of a COX-2 selective nonsteroidal anti-inflammatory drug versus placebo in inguinal herniorrhaphy patients. AB - OBJECTIVE: The standard opioid treatment for postoperative pain can be associated with nausea, vomiting, and constipation. In addition, opioids often provide insufficient pain relief. The purpose of this study was to compare postoperative pain and functional outcomes in patients undergoing inguinal herniorrhaphy who receive a COX-2 selective nonsteroidal anti-inflammatory drug (COX-2) or placebo preoperatively and for 4 days postoperatively. METHODS: A prospective, randomized, blinded, placebo-controlled trial was conducted in adults undergoing elective, outpatient, unilateral inguinal herniorrhaphy. Patients received rofecoxib (50 mg, 1 h prior to incision) or placebo. Doses were re-administered once daily on postoperative days 1-4. Patients were also given hydrocodone bitartrate for use as needed in the postoperative period. Pain outcomes were assessed, including pain intensity (1-10 visual-analogue scale) and the use of hydrocodone bitartrate. In addition, functional outcomes such as activity and return of bowel function were examined for 5 postoperative days. Incidence and severity of side effects were examined. Statistics are mean +/- standard deviation. RESULTS: Fifty-five subjects completed the study. Twenty-six patients received rofecoxib and 29 patients received placebo. Patients who received COX-2 demonstrated improved bowel function as reflected by more bowel movements on postoperative day 2 and postoperative day 3. COX-2-treated patients also reported better oral intake on these same days. In addition, COX-2-treated patients had less difficulty coughing on postoperative day 1. Overall satisfaction with pain management was better in COX-2-treated patients (very satisfied vs. satisfied). There were no statistically significant differences between groups in the amount of hydrocodone bitartrate consumption. There were no complications during the study period. CONCLUSIONS: Administration of a COX-2 selective nonsteroidal anti inflammatory drug prior to and following outpatient inguinal herniorrhaphy improves functional outcomes when compared with placebo and increases patient satisfaction. These results suggest that multimodal pain therapy with COX-2 inhibitors may have a role in outpatient inguinal hernia repair. PMID- 19301087 TI - Type 1 autoimmune hepatitis and adipokines: new markers for activity and disease progression? AB - PURPOSE: Cytokines may play an important role as inflammatory factors in liver diseases. There is some evidence suggesting a link between adiponectin-biliary function and liver disease. The aim of this study was to clarify the behavior of adipokines in autoimmune hepatitis type 1. METHODS: We assessed the circulating levels of adiponectin, tumor necrosis factor-alpha, resistin and leptin in 42 patients with autoimmune hepatitis, comparing them with 42 healthy subjects who were matched for age and sex and with 31 patients with nonalcoholic steatohepatitis (NASH), evaluating the associations with markers of cytolysis, cholestasis, and histological severity. RESULTS: Adiponectin and TNF-alpha values were higher in patients compared to controls. The patients showed significantly higher Homeostasis Model Assessment values, suggesting an increased insulin resistance and serum levels of adiponectin positively correlated with gamma glutamyltranspeptidase and alkaline phosphatase values after a simple regression analysis. Serum levels of resistin positively correlated with elevated aminotransferases and bilirubin values, and serum levels of TNF-alpha positively correlated with elevated alanine-aminotransferase and resistin values. The concentration of adiponectin increased significantly with staging of the disease. Patients with NASH showed lower levels of adiponectin and higher levels of resistin than AIH patients and controls. CONCLUSIONS: Patients with AIH showed significantly higher adiponectin concentrations than controls despite their higher HOMA-IR values. The significant correlation between adiponectin levels and serological features of cholestasis suggested an association with biliary function. Our results indicate that adiponectin may be a possible marker for disease progression in AIH. PMID- 19301088 TI - Calcification and silicification: a comparative survey of the early stages of biomineralization. AB - Most of the studies on biomineralization have focused on calcification and silicification, the two systems that predominate in nature in the construction of skeletal or integumental hard tissues. They have, however, been studied separately, as if they were completely distinct processes, in spite of their several points of contact, especially as far as the organic-inorganic relationships during the early mineralization stages are concerned. A very tight association of the inorganic substance with organic macromolecules, in fact, initially characterizes both systems. Although the mechanism of biomineralization remains elusive, a number of old and new findings, which have been taken into account in this review, support the view that, both in calcification and in silicification, genetically controlled organic macromolecules induce the formation of composite, organic-inorganic nanoparticles, behave as templates for the subsequent assemblage of the nanoparticles into micro- to macroarchitectures of complex pattern, and, eventually, are mostly reabsorbed. There are still many gaps left in our knowledge of this process. Comparative studies of the two biomineralization systems may help to fill them. PMID- 19301089 TI - The effect of risedronate treatment on serum cytokines in postmenopausal osteoporosis: a 6-month randomized and controlled study. AB - There is much evidence suggesting that the decline in ovarian function after menopause is associated with spontaneous increases in proinflammatory cytokines. Treatment with risedronate is accompanied by significant changes in bone turnover and bone mineral density. The objective of this study was to determine the effects of risedronate treatment on the level of serum cytokines including receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin among postmenopausal women with osteoporosis. The study group consisted of 61 postmenopausal women with osteoporosis. Patients were randomly divided in two groups: In group 1 (n = 41) postmenopausal women received oral risedronate (35 mg/week), calcium (1,000 mg/day), and vitamin D (400 IU/day) for 12 months. In group 2 (control group; n = 20) patients received only oral calcium (1,000 mg/day) and vitamin D (400 IU/day). Bone mineral density (BMD) of lumbar spine (L1-L4) and proximal femur were determined using dual X-ray absorptiometry at baseline and after one year. Venous blood samples were obtained for determination of serum cytokines including interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), RANKL, osteoprotegerin, and markers of bone formation and resorption. Levels of serum cytokines were measured before therapy and after three and 6 months. Markers of bone metabolism were studied before therapy and after 6 months. In group 1 (risedronate plus calcium/vitamin D-treated patients), serum levels of RANKL and IL-1beta significantly decreased and the level of osteoprotegerin significantly increased after three and 6 months, but no significant difference was found in TNF-alpha level. In group 2, however, the level of serum cytokines did not change after three and 6 months. In cases of bone turnover, both markers of bone resorption and formation significantly decreased after 6 months in group 1. In conclusion risedronate could improve osteoporosis by increasing osteoprotegerin and reducing RANKL and IL-1beta. PMID- 19301090 TI - Applications of flow cytometry in environmental microbiology and biotechnology. AB - Flow cytometry (FCM) is a technique for counting, examining and sorting microscopic particles suspended in a stream of fluid. It uses the principles of light scattering, light excitation and the emission from fluorescent molecules to generate specific multiparameter data from particles and cells. The cells are hydrodynamically focussed in a sheath solution before being intercepted by a focused light source provided by a laser. FCM has been used primarily in medical applications but is being used increasingly for the examination of individual cells from environmental samples. It has found uses in the isolation of both culturable and hitherto non-culturable bacteria present infrequently in environmental samples using appropriate growth conditions. FCM lends itself to high-throughput applications in directed evolution for the analysis of single cells or cell populations carrying mutant genes. It is also suitable for encapsulation studies where individual bacteria are compartmentalised with substrate in water-in-oil-in-water emulsions or with individual genes in transcriptional/translational mixtures for the production of mutant enzymes. The sensitivity of the technique has allowed the examination of gene optimisation by a procedure known as random or neutral drift where screening and selection is based on the retention of some predetermined level of activity through multiple rounds of mutagenesis. PMID- 19301092 TI - Quantitative analysis of fecal sapovirus shedding: identification of nucleotide substitutions in the capsid protein during prolonged excretion. AB - Sapovirus (SaV) is an important pathogen causing gastroenteritis in humans. Quantitative analysis of the viral loads in feces collected from two SaV outbreaks was performed. Our results showed that SaV excretion generally decreased to an undetectable level in 2 weeks; however, some individuals excrete SaV in feces at high concentrations for 2-4 weeks after the onset of illness. In addition, we identified for the first time nucleotide changes in the capsid region during prolonged excretion. PMID- 19301093 TI - Rapid detection and non-subjective characterisation of infectious bronchitis virus isolates using high-resolution melt curve analysis and a mathematical model. AB - Infectious bronchitis virus (IBV) is a coronavirus that causes upper respiratory, renal and/or reproductive diseases with high morbidity in poultry. Classification of IBV is important for implementation of vaccination strategies to control the disease in commercial poultry. Currently, the lengthy process of sequence analysis of the IBV S1 gene is considered the gold standard for IBV strain identification, with a high nucleotide identity (e.g. > or =95%) indicating related strains. However, this gene has a high propensity to mutate and/or undergo recombination, and alone it may not be reliable for strain identification. A real-time polymerase chain reaction (RT-PCR) combined with high resolution melt (HRM) curve analysis was developed based on the 3'UTR of IBV for rapid detection and classification of IBV from commercial poultry. HRM curves generated from 230 to 435-bp PCR products of several IBV strains were subjected to further analysis using a mathematical model also developed during this study. It was shown that a combination of HRM curve analysis and the mathematical model could reliably group 189 out of 190 comparisons of pairs of IBV strains in accordance with their 3'UTR and S1 gene identities. The newly developed RT PCR/HRM curve analysis model could detect and rapidly identify novel and vaccine related IBV strains, as confirmed by S1 gene and 3'UTR nucleotide sequences. This model is a rapid, reliable, accurate and non-subjective system for detection of IBVs in poultry flocks. PMID- 19301094 TI - Expression of mouse beta-defensin-3 in MDCK cells and its anti-influenza-virus activity. AB - Influenza (flu) pandemics have presented a threat to human health in the past century. Because of outbreaks of avian flu in humans in some developing countries in recent years, humans are more eager to find a way to control flu. Mammalian beta-defensins (beta-defensins) are associated primarily with mucosal and skin innate immunity. Previous studies have demonstrated antimicrobial properties of a variety of defensin peptides. We have identified the presence of mouse beta defensin 1, 2, and 3 genes (Mbd-1, 2, and 3) in trachea and lung tissues by RT PCR before and after infection with influenza virus. We constructed a eukaryotic expression plasmid containing Mbd-3, pcDNA 3.1(+)/MBD-3, and the plasmid was introduced into Madin-Darby canine kidney (MDCK) cells by transfection. The expression of Mbd-3 in MDCK cells was verified by immunofluorescence test, RT PCR, and Western blot. The pcDNA 3.1(+)/MBD-3 plasmid was injected into mice to observe its effect against influenza A virus (IAV) in vivo. Mouse beta-defensin genes could be expressed in trachea and lung tissues before IAV infection, but expression of Mbd-2 and Mbd-3 was increased significantly after IAV infection. The survival rate of mice with MBD-3 against IAV challenge was 71.43%, and MDCK cells with MBD-3 could clearly inhibit IAV replication. The results demonstrated that mouse beta-defensins possess anti-influenza virus activity, suggesting that mouse beta-defensins might be used as agents to prevent and treat influenza. PMID- 19301095 TI - Amino acids: metabolism, functions, and nutrition. AB - Recent years have witnessed the discovery that amino acids (AA) are not only cell signaling molecules but are also regulators of gene expression and the protein phosphorylation cascade. Additionally, AA are key precursors for syntheses of hormones and low-molecular weight nitrogenous substances with each having enormous biological importance. Physiological concentrations of AA and their metabolites (e.g., nitric oxide, polyamines, glutathione, taurine, thyroid hormones, and serotonin) are required for the functions. However, elevated levels of AA and their products (e.g., ammonia, homocysteine, and asymmetric dimethylarginine) are pathogenic factors for neurological disorders, oxidative stress, and cardiovascular disease. Thus, an optimal balance among AA in the diet and circulation is crucial for whole body homeostasis. There is growing recognition that besides their role as building blocks of proteins and polypeptides, some AA regulate key metabolic pathways that are necessary for maintenance, growth, reproduction, and immunity. They are called functional AA, which include arginine, cysteine, glutamine, leucine, proline, and tryptophan. Dietary supplementation with one or a mixture of these AA may be beneficial for (1) ameliorating health problems at various stages of the life cycle (e.g., fetal growth restriction, neonatal morbidity and mortality, weaning-associated intestinal dysfunction and wasting syndrome, obesity, diabetes, cardiovascular disease, the metabolic syndrome, and infertility); (2) optimizing efficiency of metabolic transformations to enhance muscle growth, milk production, egg and meat quality and athletic performance, while preventing excess fat deposition and reducing adiposity. Thus, AA have important functions in both nutrition and health. PMID- 19301096 TI - RBL-2H3 cells are an imprecise model for mast cell mediator release. AB - OBJECTIVE: The cell line, RBL-2H3, has been widely used as a mast cell model though much of the data is contradictory. The aim of this study is to assess the RBL-2H3 cell line as an in vitro model for degranulation studies. METHODS: RBL 2H3 cells were stimulated with either dinitrophenylated-IgE, calcium ionophore A23187, compound 48/80, mast cell degranulating peptide or lipopolysaccharide and mediator (histamine, beta-hexosaminidase, interleukin-13 and TNF-alpha) release was analysed. Toll-like receptors (TLR) mRNA expression in RBL-2H3 cells and rat peritoneal mast cells (RPMC) were compared by RT-PCR. TLR4 and CD14 surface proteins in RBL-2H3 were analysed by FACS. RESULTS: Mediator release was dependent on media composition and degranulation was stimulated by IgE and A23187. Degranulation was inhibited by quercetin, but not by cromoglycate or ketotifen. Transcripts encoding TLR3-5 and 6 were detected in RBL-2H3 cells whereas TLR1-6 and TLR8 were clearly seen in RPMC. While proteins were detected for TLR4 in RBL-2H3 cells, CD14 was largely absent. CONCLUSIONS: While RBL-2H3 cells may be useful as a model for mast cell IgE-mediated degranulation, other aspects may not be representative and they may share similarities with basophils rather than with other histamine-releasing cell types. PMID- 19301097 TI - MAP kinases in proliferating human colon cancer Caco-2 cells. AB - The mitogen-activated protein kinase (MAPK) cascade is one of the most ubiquitous signal transduction systems and is rapidly activated by various stimuli, such as cellular stress and death. The Caco-2 cell line is an in vitro model for colon cancer studies. We investigated the activation status of the ERK1/2, p38, JNK1/2, and ERK5 kinases and their respective upstream intracellular activators in Caco-2 cells induced to proliferate by 10% fetal bovine serum (FBS). The states of phosphorylation of the above MAPKs and their upstream kinases, MEK1/2, MKK3/6, MKK4, and MKK7, respectively, were studied by Western blot analysis. Phosphorylation was barely detectable before serum stimulation, and the stimulation of cell proliferation by the addition of FBS increased MEK1/2 and ERK1/2 phosphorylation 2 to 3 fold after 3 min. FBS stimulated p38 and MKK3/6 to the same extent within 2 min of treatment and JNK1/2 and its upstream kinases MKK4 and MKK7 5-fold (3 min). Addition of FBS also rapidly phosphorylated ERK5 (2 to 3.5-fold between 2 and 5 min) and the transcription factor CREB. Incubation of Caco-2 cells with FBS was followed by a rapid induction of c-Fos and c-Myc expression. Studies with ERK1/2 specific inhibitor PD98059, p38 MAPK inhibitor SB203580, or JNK inhibitor SP600125 showed that FBS regulates Caco-2 cell proliferation via the three MAPK pathways. PMID- 19301098 TI - Hippocampal neurodegeneration in experimental autoimmune encephalomyelitis (EAE): potential role of inflammation activated myeloperoxidase. AB - Experimental Autoimmune Encephalomyelitis (EAE) is a well-established animal model of human multiple sclerosis (MS). The effect of this inflammatory disease on hippocampus has not been addressed. Keeping in view the above consideration an attempt was made to delineate the effect of EAE on the hippocampus of Wistar rats. The assessment of the damage to the hippocampus was done 16 days post induction by the immunolocalization of ChAT (choline acetyl transferase). ChAT decreased remarkably after induction that revealed cholinergic neuronal degeneration in the hippocampus. Subsequently, many biochemical parameters were assessed to ascertain inflammatory activation of nitric oxide and associated oxidative damage as a putative mechanism of the cholinergic degeneration. Nitric oxide metabolites increased significantly (P < 0.05) with enhancement of MPO (Myeloperoxidase activity) (P < 0.001) in the MOG (myelin oligodendrocyte protein) group as compared to the controls. Peroxidation of biomembranes increased (P < 0.001), while reduced glutathione depleted (P < 0.001) with parallel decrease in catalase (P < 0.01) and superoxide dismutase enzyme activity (P < 0.001) in the MOG group. Our results show a strong role of peroxidase dependent oxidation of nitrite and oxidative stress in cholinergic degeneration in EAE. PMID- 19301099 TI - Evaluation of different methods detecting intracellular generation of free radicals. AB - Reactive oxygen species (ROS) play several biological roles. We investigated the applicability of fluorescent probes for their detection (i) in rabbit lens epithelial cells during ageing in culture, and (ii) in thin sections of rat heart. We used dihydroethidium (DHE), dichlorofluorescin (DCFH), and dihydrorhodamine 123 (DHR) together with detection of autofluorescence both in cells and in chloroform extracts. Superoxide production was confirmed by a specific histochemical method using Mn(2+). All methods demonstrated higher production of ROS in older cells. All probes revealed different sites of ROS production in young and old cells and could be used for investigation of ROS generation during cell ageing. In the thin sections of rat heart DCFH was not suitable for intracellular ROS detection. The results indicate that the potential of fluorescent dyes in ROS detection is not usually fully exploited, and that blue autofluorescence is associated with oxidative damage. PMID- 19301100 TI - Stimulation of adenosine A2b receptors blocks apoptosis in the non-infarcted myocardium even when administered after the onset of infarction. AB - OBJECTIVE: Chronic adenosine A2b receptor stimulation has been shown to prevent ventricular remodelling after myocardial infarction (MI). We hypothesized that this effect is due to the inhibition of cardiac myocyte apoptosis in the myocardium remote from the infarction. METHODS: Rats were subjected to MI by LAD ligation in situ. Some animals were pre-treated with the stable adenosine analogue 2-chloro-adenosine (CADO). After 24 h, pro- and anti-apoptotic signals (protein kinase C isoforms, p38, g proteins, Bcl-2/Bax ratio, Akt, Bad), and marker of apoptosis execution (caspase-3, TUNEL) were quantified in the remote myocardium. RESULTS: CADO prevented the occurrence of apoptosis in the remote myocardium of an infarcted heart. This effect occured not only when CADO was started before the onset of ischemia but also when it started 3 h after the infarction. The anti-apoptotic effect of CADO was blocked by simultaneous administration of the selective adenosine A2b receptor antagonist MRS1754 (1 mg/kg). The anti-apoptotic effect of CADO seems to be mediated by g(alphaq) and by the activation of survival kinases (Bad) and by inhibition of the pro apoptotic PKC-delta/p38-MAPK-pathway. CONCLUSION: Chronic adenosine A2b receptor stimulation blocks cardiac myocyte apoptosis in the remote myocardium even when started after the onset of infarction. This may explain the anti-remodelling effect of the A2b receptor stimulation after infarction. PMID- 19301101 TI - Human tissue allograft processing: impact on in vitro and in vivo biocompatibility. AB - This work investigates the impact of chemical and physical treatments on biocompatibility for human bone/tendon tissues. Nontreated and treated tissues were compared. In vitro testing assessed indirect and direct cytotoxicity. Tissues were subcutaneously implanted in rats to assess the immunological, recolonization, and revascularization processes at 2-4 weeks postimplantation. No significant cytotoxicity was found for freeze-dried treated bones and tendons in comparison to control. The cellular adhesion was significantly reduced for cells seeded on these treated tissues after 24 h of direct contact. A significant cytotoxicity was found for frozen treated bones in comparison to freeze-dried treated bones. Tissue remodeling with graft stability, no harmful inflammation, and neo-vascularization was observed for freeze-dried chemically treated bones and tendons. Frozen-treated bones were characterized by a lack of matrix recolonization at 4 weeks postimplantation. In conclusion, chemical processing with freeze-drying of human tissues maintains in vitro biocompatibility and in vivo tissue remodeling for clinical application. PMID- 19301102 TI - Phase assemblage study and cytocompatibility property of heat treated potassium magnesium phosphate-silicate ceramics. AB - This article reports the study on a new generation bioactive ceramic, based on MgKPO(4) (Magnesium Potassium Phosphate, abbreviated as MKP) for biomedical applications. A series of heat treatment experiments on the slip cast silica (SiO(2)) containing MKP ceramics were carried out at 900, 1,000 and 1,100 degrees C for 4 h in air. The density of the slip cast ceramic increases to 2.5 gm/cm(3) upon heat treatment at 900 degrees C. However, no significant change in density is measured upon heat treatment to higher temperature of 1,000 and 1,100 degrees C. On the basis of XRD results, the presence of K(2)MgSi(5)O(12) and dehydrated MgKPO(4) were confirmed and complementary information has also been obtained using FT-IR and Raman spectroscopy. In order to confirm the in vitro cytocompatibility property, the cell culture tests were carried out on selected samples and the results reveal good cell adhesion and spreading of L929 mouse fibroblast cells. MTT assay analysis with L929 cells confirmed non-cytotoxic behavior of MKP containing ceramics and the results are comparable with sintered HAp ceramics. It is expected that the newly developed MKP based materials could be a good substitute for hydroxyapatite (HAp or HA) based bioceramics. PMID- 19301103 TI - Silicon-substituted hydroxyapatite composite coating by using vacuum-plasma spraying and its interaction with human serum albumin. AB - The incorporation of silicon can improve the bioactivity of hydroxyapatite (HA). Silicon-substituted HA (Ca(10)(PO(4))(6-x )(SiO(4))( x )(OH)(2-x ), Si-HA) composite coatings on a bioactive titanium substrate were prepared by using a vacuum-plasma spraying method. The surface structure was characterized by using XRD, SEM, XRF, EDS and FTIR. The bond strength of the coating was investigated and XRD patterns showed that Ti/Si-HA coatings were similar to patterns seen for HA. The only different XRD pattern was a slight trend toward a smaller angle direction with an increase in the molar ratio of silicon. FTIR spectra showed that the most notable effect of silicon substitution was that -OH group decreased as the silicon content increased. XRD and EDS elemental analysis indicated that the content of silicon in the coating was consistent with the silicon-substituted hydroxyapatite used in spraying. A bioactive TiO(2) coating was formed on an etched surface of Ti, and the etching might improve the bond strength of the coatings. The interaction of the Ti/Si-HA coating with human serum albumin (HSA) was much greater than that of the Ti/HA coating. This might suggest that the incorporation of silicon in HA can lead to significant improvements in the bioactive performance of HA. PMID- 19301104 TI - Microporous biodegradable polyurethane membranes for tissue engineering. AB - Microporous membranes with controlled pore size and structure were produced from biodegradable polyurethane based on aliphatic diisocyanate, poly(epsilon caprolactone) diol and isosorbide chain extender using the modified phase inversion technique. The following parameters affecting the process of membrane formation were investigated: the type of solvent, solvent-nonsolvent ratio, polymer concentration in solution, polymer solidification time, and the thickness of the polymer solution layer cast on a substrate. The experimental systems evaluated were polymer-N,N-dimethylformamide-water, polymer-N,N-dimethylacetamide water and polymer-dimethylsulfoxide-water. From all three systems evaluated the best results were obtained for the system polymer-N,N-dimethylformamide-water. The optimal conditions for the preparation of microporous polyurethane membranes were: polymer concentration in solution 5% (w/v), the amount of nonsolvent 10% (v/v), the cast temperature 23 degrees C, and polymer solidification time in the range of 24-48 h depending on the thickness of the cast polymer solution layer. Membranes obtained under these conditions had interconnected pores, well defined pore size and structure, good water permeability and satisfactory mechanical properties to allow for suturing. Potential applications of these membranes are skin wound cover and, in combination with autogenous chondrocytes, as an "artificial periosteum" in the treatment of articular cartilage defects. PMID- 19301105 TI - A novel composite membrane of chitosan-carboxymethyl cellulose polyelectrolyte complex membrane filled with nano-hydroxyapatite I. Preparation and properties. AB - A novel tri-component composite membranes of chitosan/carboxymethyl cellulose (CS/CMC) polyelectrolyte complex membranes filled with different weight ratios of nano-hydroxyapatite (n-HA)(0, 20, 40 and 60 wt%), namely, n-HA/CS/CMC composite membrane, were prepared by self-assembly of static electricity. The structure and the properties of the composite membranes were investigated by Fourier transformed infrared spectroscopy(IR), X-ray diffraction(XRD), Scanning electron microscopy(SEM), mechanical performance measurement, swelling behavior test, and soaking behavior study in phosphate buffered saline (PBS) and simulate body fluid (SBF). The results showed that the n-HA/CS/CMC composite membrane was formed though superficial static electricity interaction among n-HA, CS and CMC. For the n-HA/CS/CMC composite membrane, the microstructure compatibility, mechanical property, swelling behavior, the degradation and bioactivity in vitro of the composite membrane were improved by the addition of n-HA, compared with CS/CMC polyelectrolyte complex membrane. Moreover, the n-HA/CS/CMC composite membrane with 40 wt% n-HA had the most highest mechanical property, which suggested that the novel n-HA/CS/CMC composite membrane with 40 wt% n-HA was more suitable to be used as guided bone tissue regeneration membrane than CS/CMC polyelectrolyte complex membrane. PMID- 19301106 TI - Influence of fabrication parameters in cellular microarrays for stem cell studies. AB - Lately there has been an increasing interest in the development of tools that enable the high throughput analysis of combinations of surface-immobilized signaling factors and which examine their effect on stem cell biology and differentiation. These surface-immobilized factors function as artificial microenvironments that can be ordered in a microarray format. These microarrays could be useful for applications such as the study of stem cell biology to get a deeper understanding of their differentiation process. Here, the evaluation of several key process parameters affecting the cellular microarray fabrication is reported in terms of its effects on the mesenchymal stem cell culture time on these microarrays. Substrate and protein solution requirements, passivation strategies and cell culture conditions are investigated. The results described in this article serve as a basis for the future development of cellular microarrays aiming to provide a deeper understanding of the stem cell differentiation process. PMID- 19301107 TI - In vitro biocompatibility of chitosan-based materials to primary culture of hippocampal neurons. AB - The natural biomaterial chitosan has been widely used as a promising nerve guidance conduit material for peripheral nerve repair. This study aimed to investigate in vitro biocompatibility of chitosan to primarily cultured hippocampal neurons, one type of central nervous system (CNS) cells. The substrate made up of chitosan fibers or membranes was found to support the survival and growth of the attached hippocampal neurons by using light and electron microscopy as well as immunocytochemistry for neurofilament 200, growth associated protein-43, microtubule-associated protein 2, beta-tubulin III and synaptophysin. MTT assay indicated that the cell viability of hippocampal neurons in chitosan fiber or membrane extract was not significantly different from that in hydroxyapatite extract or plain neuronal medium, but significantly higher than that in organotin extract after culture for different times. Western analysis revealed that no significant difference in the protein level of growth-associated protein-43 and beta-tubulin III was detected between hippocampal neurons cultured in chitosan extract and in plain neuronal culture medium. The results collectively demonstrate that chitosan is biocompatible to primary culture of hippocampal neurons without cytotoxic effects on cell phenotype and functions, raising a potential possibility of using chitosan for CNS therapy. PMID- 19301108 TI - Comparison of the psychometric properties of four at-work disability measures in workers with shoulder or elbow disorders. AB - INTRODUCTION: To better capture the extent of work disability following an occupational injury, clinical researchers are beginning to recognize the importance of considering not only levels of absenteeism, but also disabilities experienced while "at-work". Although at-work disability measures are available in the literature, currently there is little insight on the selection of specific measures that may be best suited for a given population or situation. The objective of this study is to assess and compare the measurement properties of four self-report at-work disability measures in workers with shoulder or elbow disorders. METHODS: Study sample consisted of 80 patients attending a shoulder and elbow specialty clinic operated by the Worker Safety Insurance Board of Ontario. Internal consistency reliability, validity, and patient preference of four at-work disability measures were compared in a cross-sectional design. Selected measures included the work module of the Disabilities of the Arm, Shoulder and Hand Outcome Measure, Work Limitations Questionnaire (WLQ-16), Work Instability Scale for Rheumatoid Arthritis (RA-WIS), and Stanford Presenteeism Scale. RESULTS: All four measures demonstrated evidence of internal consistency reliability (alpha = 0.76-0.90) and construct validity, although only modest correlations against work-oriented constructs (r = 0.37-0.60) were observed. The RA-WIS was most preferred by respondents (44.6%) over the other measures. CONCLUSIONS: Although no single scale stood out as clearly superior, the WLQ-16 was considered the best overall performer. Variable performance between the scales suggests some divergence in the way these measures conceptualize "at-work disability", which may be important to consider when selecting instruments for future studies. PMID- 19301109 TI - An investigation of relationships among genetic counselors' supervision skills and multicultural counseling competence. AB - As racial and ethnic diversity increase in the U.S., genetic counselor multicultural competence is growing in importance. In mental health counseling, supervisor multicultural competence has been shown to promote supervisees' multicultural competence. Moreover, developmentally-advanced supervisors tend to be more effective. This study was designed to investigate relationships among genetic counselor supervisors' perceived multicultural counseling competence and development as supervisors, and their ability to evaluate a supervisee's multicultural skills. One hundred twenty-two supervisors completed an online survey of demographics, the Multicultural Counseling Knowledge and Awareness Scale, the Supervisor Development Scale, and a hypothetical vignette in which they evaluated a supervisee's multicultural skills and provided written feedback. Stepwise multiple regression yielded five significant predictors accounting for 31% of the variance in accuracy of supervisor evaluations of the student: multicultural awareness, multicultural knowledge, age, supervision experience, and supervisor development. Six feedback themes were identified from written responses. Practice and research suggestions are provided. PMID- 19301110 TI - Inoculation or antidote? The effects of cognitive interview timing on false memory for forcibly fabricated events. AB - This study examined whether a cognitive interview (CI) can counteract the effects of suggestive interviews involving forced fabrication. College students witnessed a filmed event and were later forced to fabricate answers to misleading questions about the event. All witnesses were interviewed with a non-leading CI or free recall (FR) either before or after the forced fabrication phase. A week later participants completed a recognition and source monitoring (SM) test of video content. Relative to FR, the CI administered before the forced fabrication interview increased reports of correct details and reduced false assents to fabricated items. A CI after resulted in false memory rates comparable to the FR group. Early interviews using CI techniques may protect against memory loss and misinformation effects. PMID- 19301111 TI - Etanercept impairs maturation of human monocyte-derived dendritic cells by inhibiting the autocrine TNFalpha-mediated signaling. AB - The success of anti-tumor necrosis factor alpha (TNFalpha) therapies has led to increased interest as to the mechanisms and consequences of TNFalpha blockade. The aim of the study was to examine the effects of TNFalpha blockade by etanercept on lipopolysaccharide (LPS) or peptidoglycan (PG)-induced maturation of human monocyte-derived dendritic cells (MDDCs). MDDCs grown from peripheral blood of healthy donors were stimulated by LPS or PG with/without the presence of etanercept. Concentrations of TNFalpha in cell supernatants were assessed by ELISA, while the cells were stained with monoclonal antibodies to CD83, CD80, CD86, CD11c, CD40, HLA-DR, and annexin-V and acquired using a flow cytometer. Etanercept significantly decreased the stimulated cell surface expression of HLA DR, CD80, CD86, CD40 and CD83 on MDDCs in all examined samples. Etanercept in the same dose, but denatured to loss of specificity for TNFalpha, failed to change any of the aforementioned markers. In the presence of etanercept, concentrations of TNFalpha in cell supernatants were decreased by 53% on average, with a range of 25%-87%. Etanercept impaired the stimulated maturation of MDDCs by neutralizing the induced TNFalpha, produced by the same MDDCs after antigenic stimulation. The reported data confirms that TNFalpha blockade may have a direct effect on DCs, with a wide spectrum of potential secondary effects downstream. The data also suggests the presence of TNFalpha-mediated autocrine signaling, serving to accelerate or catalyze the maturation process of MDDCs. PMID- 19301112 TI - Sexual dimorphism in the prenatal digit ratio (2D:4D). AB - The second to fourth digit ratio (2D:4D) is smaller in human males than in females and hence this trait is sexually dimorphic. The digit ratio is thought to be established during early prenatal development under the influence of prenatal sex hormones. However, the general assumption of early establishment has hardly been studied. In our study, we analyzed the 2D:4D ratio in 327 deceased human fetuses. We measured digit lengths in 169 male and 158 female fetuses ranging from 14 to 42 weeks old. Our results showed a slight, but significant, sexual dimorphism in the expected direction, i.e., females had, on average, a ratio of 0.924 and males a ratio of 0.916. There was no significant relationship with the presence or absence of minor and major or single and multiple congenital abnormalities. There was a minimal, but significant difference between digit ratios based on digit lengths including and excluding the non-bony fingertip with the values being strongly correlated (r = .98). The prenatal 2D:4D ratio was lower than has thus far been reported for children and adults both for males and females. The extent of the sexual dimorphism in fetuses was similar to that found for children, but lower than for adults. The 2D:4D ratio, thus, seems to increase after birth in both men and women, with the second digit growing faster than the fourth digit (positive allometric growth of digit two) and perhaps more so in women than in men. Therefore, the sexual dimorphism is probably determined by prenatal as well as by postnatal developmental processes. PMID- 19301113 TI - Longitudinal antiretroviral adherence in HIV+ Ugandan parents and their children initiating HAART in the MTCT-Plus family treatment model: role of depression in declining adherence over time. AB - We conducted a study to assess the effect of family-based treatment on adherence amongst HIV-infected parents and their HIV-infected children attending the Mother To-Child-Transmission Plus program in Kampala, Uganda. Adherence was assessed using home-based pill counts and self-report. Mean adherence was over 94%. Depression was associated with incomplete adherence on multivariable analysis. Adherence declined over time. Qualitative interviews revealed lack of transportation money, stigma, clinical response to therapy, drug packaging, and cost of therapy may impact adherence. Our results indicate that providing ART to all eligible HIV-infected members in a household is associated with excellent adherence in both parents and children. Adherence to ART among new parents declines over time, even when patients receive treatment at no cost. Depression should be addressed as a potential barrier to adherence. Further study is necessary to assess the long-term impact of this family treatment model on adherence to ART in resource-limited settings. PMID- 19301114 TI - Women living with HIV in South Africa and their concerns about fertility. AB - Health and quality of life benefits accrued from the availability of highly active antiretroviral therapy (HAART) are commendable, but the social milieu continues to pose challenges for women's decision making around having children. This paper qualitatively explored women's questions and concerns around living with HIV, being on HAART and pregnancy. Women of reproductive age were recruited from Eastern Cape and Gauteng Provinces, South Africa. Information on women's fertility desires and pregnancy planning was collected through participatory workshop, focus groups, and one-on-one interviews. Three main themes emerged. Women living with HIV require information on the impact of HIV on pregnancy outcomes and vice versa. Women who are young, lost a child, not consistently using contraception or who have not been seriously ill have positive reproductive aspirations. Ambivalent attitudes of health care workers towards pregnancy impacts women's fertility aspirations. Unbiased pre-conceptual communication should form part of HIV treatment and care services, despite expressed pregnancy intentions. PMID- 19301115 TI - Responding to her question: a review of the influence of pregnancy on HIV disease progression in the context of expanded access to HAART in sub-Saharan Africa. AB - In 2007, sub-Saharan Africa was home to over half of all women living with HIV. The vast majority of these women are of reproductive age, which raises concerns about the high incidence of pregnancy. As access to antiretroviral treatment is rapidly scaled up, two important questions must be answered: (1) Does pregnancy impact HIV disease progression?; (2) Does pregnancy modify the highly active antiretroviral therapy (HAART) response on HIV disease progression? A systematic review of the biomedical literature was conducted and seven relevant studies were identified. To date, it appears that there is no effect of pregnancy on HIV disease progression. Furthermore, initial studies in high-income countries suggest that pregnancy may positively modify the HAART response. These findings, however, must be interpreted with caution as it remains unclear how other factors, such as adherence, may influence the relationship between pregnancy, HIV disease progression, and HAART. PMID- 19301116 TI - The impact of HIV status and perceived status on fertility desires in rural Malawi. AB - Surprisingly little is known about how or if knowledge of one's HIV status influences the desire to have children in sub-Saharan Africa. This paper takes advantage of a unique situation in rural Malawi where no one knew their HIV status prior to testing being introduced as part of an ongoing panel study. Using a sample of men and women (n = 1,380) who were interviewed in 2001 and 2006, we examine how HIV positive and negative test results impact respondents' desires to continue childbearing. We then consider how prior perceptions of one's status influence this effect. Respondents who received a positive test result reduced their childbearing desires. When self-assessed likelihood of infection was considered, however, only those who were surprised by their test result altered their fertility preferences, a finding which held true for both positive and negative results. The implications of the results and potential applications to other HIV/AIDS research are discussed. PMID- 19301117 TI - Early pregnancy loss rates were different among singleton gestations conceived by ICSI using GnRH agonist and antagonist. AB - This study compared early pregnancy losses (termination of pregnancy before 12 weeks of gestation, EPL) among conceptions achieved by ICSI according to the type of GnRH analogue for ovarian stimulation. Only singleton gestations (2,184) and fresh embryo transfers were included. GnRH agonist was used in 848 gestations out of 2,184 and GnRH antagonist was used in the remaining 1,336 gestations. EPL was found to be significantly higher in GnRH antagonist gestations compared to GnRH agonist (27.2% vs 18.9%). This significant difference persisted when gestations were segregated according to maternal age, especially among women younger than 35 years old. Therefore our results suggest that gestations conceived by ovarian stimulation including GnRH antagonists may have higher probability of having EPL. PMID- 19301118 TI - Mediating and moderating processes in the relation between maltreatment and psychopathology: mother-child relationship quality and emotion regulation. AB - The present study investigated underlying processes of the effect of maltreatment on psychopathology (i.e., internalizing and externalizing problems) in a group of 111 maltreated and 110 nonmaltreated 7-10 year-old children (60% boys). We tested the moderating and/or mediating roles of emotion regulation and the mother-child relationship quality (pattern of relatedness) using Structural Equation Modeling. Emotion regulation, but not the pattern of relatedness, mediated the relation between maltreatment and psychopathology. This mediation was moderated by the pattern of relatedness: For the group of children with an insecure pattern of relatedness, maltreatment was related to lower levels of emotion regulation, which was predictive of higher levels of internalizing and externalizing symptomatology. In contrast, for the secure relatedness group, there was no mediation by emotion regulation since the impact of maltreatment on emotion regulation was not significant. Implications of the mediating role of emotion regulation and the buffering role of the mother-child relationship quality were discussed. PMID- 19301119 TI - Accuracy of self-reported breast cancer among women undergoing mammography. AB - This study estimated the sensitivity and specificity of self-reported breast cancer and their associations with patient factors and pathologic findings using data from the Breast Cancer Surveillance Consortium. We included 24,631 women with and 463,804 women without a prior diagnosis of breast cancer who completed a questionnaire (including breast cancer history) at participating US mammography facilities between 1996 and 2006. We determined "true" cancer status using cancer registries and pathology databases. Multivariable logistic regression models were used to examine associations with patient factors and pathologic findings. Sensitivity of self-reported breast cancer was higher for women with invasive cancer (96.9%) than for those with ductal carcinoma in situ (DCIS) (90.2%). Specificity was high overall (99.7%) but much lower for women with a history of lobular carcinoma in situ (LCIS) (65.0%). In multivariable models, women reporting older ages, a nonwhite race/ethnicity, or less education had lower sensitivities and specificities. Sensitivity was reduced when there was evidence of prior DCIS, especially when this diagnosis had been made more than 2 years before questionnaire completion. Women reporting a family history of breast cancer had higher sensitivity. Evidence of prior LCIS was associated with lower specificity. The accuracy of self-reported breast cancer depends on the respondent's characteristics and prior diagnoses. Accuracy is lower among nonwhite women and women reporting less education. There appears to be uncertainty surrounding breast findings such as DCIS and LCIS. These results have important implications for research relying on self-report and for patient communication and care. PMID- 19301120 TI - Effects of autotransplanted lymph node fragments on the lymphatic system in the pig model. AB - Secondary lymphedema often develops after removal of lymph nodes in combination with radiation therapy, in particular in patients with breast cancer, inguinal cancer, cervical cancer and melanoma. No convincing treatment for the prevention and therapy of acquired lymphedema exists so far, therefore we wanted to show the reintegration of transplanted avascular lymph node fragments in the lymphatic system and positive effects of the transplanted fragments on the restoration of the lymphatic flow in this study. A total of 26 minipigs underwent lymphadenectomy of both groins. A minimum of one lymph node was retransplanted. The lymph nodes were cut into small pieces and retransplanted in the left groin (n = 17) or in both groins (n = 9). Different retransplantation techniques were investigated, transplantation of large versus small fragments, with and without capsule. The lymph flow was evaluated 5 and 8 months after surgery, using SPECT/CT and Berlin Blue. The results were confirmed by dissection. The lymph node transplants were assessed histologically. In contrast to the lymph flow in the transplanted groin, the lymph flow in the non-transplanted groin was often malfunctioning. Large lymph node fragments were found reintegrated in the lymphatic system more often than small slices of lymph node fragments. About 5 months after surgery impairment of lymph flow was seen especially after retransplantation of small slices of lymph node fragments. In seven out of eight minipigs a dermal backflow developed in the non-transplanted groin, 8 months after surgery. Only one minipig of these groups developed dermal backflow in both groins. All lymph node fragments showed an organized structure histologically. Autologous lymph node transplantation has positive effects on the regeneration of lymph vessels and restoration of lymph flow after lymphadenectomy. PMID- 19301121 TI - Both t-Darpp and DARPP-32 can cause resistance to trastuzumab in breast cancer cells and are frequently expressed in primary breast cancers. AB - The clinical use of trastuzumab (Herceptin), a humanized antibody against the HER2 growth factor receptor, has improved survival in patients with breast tumors with ERBB2 amplification and/or over-expression. However, most patients with advanced ERBB2 amplified breast cancers whose tumors initially respond to trastuzumab develop resistance to the drug, leading to tumor progression. To identify factors responsible for acquired resistance to trastuzumab, gene expression profiling was performed on subclones of an ERBB2 amplified breast cancer cell line, BT474, which had acquired resistance to trastuzumab. The most overexpressed gene in these subclones was PPP1R1B, encoding the DARPP-32 phosphatase inhibitor. Western analysis revealed that only the truncated isoform of the DARPP-32 protein, t-Darpp, was overexpressed in the trastuzumab resistant cells. Using gene silencing experiments, we confirmed that t-Darpp over expression was required for trastuzumab resistance in these cells. Furthermore, transfecting t-Darpp in parental BT-474 cells conferred resistance to trastuzumab, suggesting that t-Darpp expression was sufficient for trastuzumab resistance. We also found that t-Darpp over-expression was associated with Akt activation and that the T75 residue in t-Darpp was required for both Akt activation and trastuzumab resistance. Finally, we found that full-length DARPP 32 and t-Darpp are expressed in a majority of primary breast tumors. Over expression of full-length DARPP-32 can also confer resistance to trastuzumab and, moreover, is associated with a poor prognostic value in breast cancers. Thus, t Darpp and DARPP-32 expression are novel prognostic and predictive biomarkers in breast cancer. PMID- 19301122 TI - Sox17, the canonical Wnt antagonist, is epigenetically inactivated by promoter methylation in human breast cancer. AB - SRY-box 17 (Sox17) is a transcription factor which involved in a variety of developmental processes and can act as an antagonist of canonical Wnt/beta catenin signaling pathway. However, the relationship between Sox17 gene expression, methylation status, and beta-catenin in breast cancer has not been established. Here we report that the expression level of Sox17 mRNA was dramatically decreased in five different breast cancer cell lines and 23 of 31 primary breast tumor samples, which significantly correlated with its methylation status. After treated with 5-aza-2'-deoxycytidine (5-aza-dC, a demethylation agent), the expression levels of Sox17 mRNA and protein were obviously increased. Restored expression of Sox17 by 5-aza-dC treatment decreased the expression level of beta-catenin in breast cancer cell lines. Furthermore, small interfering RNA (siRNA)-mediated knockdown of Sox17 in SKBR-3 and Bacp-37 cells enhanced beta catenin expression. In 31 paired tissue samples, a significant difference between the expression level of Sox17 and beta-catenin was also observed (P < 0.001). Clinically, Sox17 methylation was detected in 74.3% breast tumors (84/113) and 31.9% (36/113) paired normal tissues, respectively (P < 0.0001). Sox17 methylation was also associated with tumor stage (P = 0.028) and lymph node metastasis (P = 0.013). These findings indicate that silencing of Sox17 due to promoter hypermethylation is a frequent event and may contribute to aberrant activation of Wnt signaling in breast cancer. Sox17 may be a valuable biomarker for the study of breast cancer carcinogenesis and progression. PMID- 19301123 TI - Population ageing and its implications on aggregate health care demand: empirical evidence from 22 OECD countries. AB - Recent evidence indicates that the relationship between age and health care expenditure is not as straightforward as it appears. In fact, micro-level studies find that time to death, rather than ageing, is possibly the main driver of the escalating health care costs in developed countries. Unfortunately, the evidence at the macro level is less clear and often depends on the specification of the empirical model used. We use an aggregate demand framework to assess whether health expenditure is more likely to be driven by ageing per se or proximity to death. Using panel data from 22 OECD countries from the first half of the 1990s, we find population ageing to be negatively correlated with health expenditure once proximity to death is accounted for. This suggests that the effects of ageing on health expenditure growth might be overstated while the effects of the high costs of medical care at the end of life are potentially underestimated. With respect to the latter, our finding highlights the importance of long-term and hospice care management. An expanded long-term care program may not only improve patient welfare, but also reduce costs of care by reducing the duration of hospital care for terminally ill patients. If expensive medical treatment for patients near the end of life can be controlled for, health expenditure growth resulting from population ageing is unlikely to present a most serious problem. PMID- 19301124 TI - Dietary enrichment with medium chain triglycerides (AC-1203) elevates polyunsaturated fatty acids in the parietal cortex of aged dogs: implications for treating age-related cognitive decline. AB - Dogs demonstrate an age-related cognitive decline, which may be related to a decrease in the concentration of omega-3 polyunsaturated fatty acids (n-3 PUFA) in the brain. Medium chain triglycerides (MCT) increase fatty acid oxidation, and it has been suggested that this may raise brain n-3 PUFA levels by increasing mobilization of n-3 PUFA from adipose tissue to the brain. The goal of the present study was to determine whether dietary MCT would raise n-3 PUFA concentrations in the brains of aged dogs. Eight Beagle dogs were randomized to a control diet (n = 4) or an MCT (AC-1203) enriched diet (n = 4) for 2 months. The animals were then euthanized and the parietal cortex was removed for phospholipid, cholesterol and fatty acid determinations by gas-chromatography. Dietary enrichment with MCT (AC-1203) resulted in a significant increase in brain phospholipid and total lipid concentrations (P < 0.05). In particular, n-3 PUFA within the phospholipid, unesterified fatty acid, and total lipid fractions were elevated in AC-1203 treated subjects as compared to controls (P < 0.05). Brain cholesterol concentrations did not differ significantly between the groups (P > 0.05). These results indicate that dietary enrichment with MCT, raises n-3 PUFA concentrations in the parietal cortex of aged dogs. PMID- 19301126 TI - Feasibility of MRI in experimentally induced inflammatory small bowel disease: a pilot study in a porcine model. AB - The purpose of this study was to compare the macroscopic and microscopic findings of experimentally induced inflammatory lesions in jejunum and ileum with magnetic resonance imaging (MRI) findings. Inflammatory small bowel lesions were experimentally induced in six pigs. Bowel segments in jejunum and ileum were isolated, and a solution with trinitrobenzenesulfonic acid and ethanol (TNBS EtOH) was installed. MRI of the small bowel was performed 7 days after surgery. Before the MRI examination, a 6% mannitol solution was installed through a nasogastric tube. The MRI protocol consisted of single-shot turbo spin echo T2 sequences, steady state free precession (BFFE) sequences, and a 3D T1 gradient echo sequence with fat saturation and intravenous contrast. The following image findings were evaluated: increased bowel wall thickness (BWT), increased bowel wall enhancement (BWE), and bowel stenosis. After the MRI examination, the animals were sacrificed. The small bowel was removed and examined macroscopically and microscopically. Inflammatory lesions developed in jejunum and ileum in all animals. The lesions were visible macroscopically and microscopically. The microscopic findings consisted of variable degrees of inflammation, ulcer formation, and fibrosis. In jejunum the inflammatory lesions were not diagnosed with MRI, except in one pig with a bowel necrosis probably caused by an intramural injection or leakage of the TNBS-EtOH solution. In ileum the bowel wall thickness was increased and the inflammatory lesions were diagnosed with MRI. In conclusion, the inflammatory lesions were visible macroscopically and microscopically. Lesions in ileum had increased BWT and were possible to image with MRI. Lesions in jejunum had normal BWT and were not diagnosed with MRI, except in one pig with increased BWT probably caused by complications to the installation of TNBS-EtOH. PMID- 19301127 TI - Variability of esterase patterns in adult flies of the saltans species group of Drosophila (subgenus Sophophora). AB - Esterases are known for their involvement in several physiological processes and high degree of polymorphism, in many organisms. Such polymorphism has been used to characterize species and species groups and to study genetic changes occurred in their evolutionary history. In the present study, the esterase patterns of 19 strains from 10 species representative of the five subgroups of the saltans species group were analyzed using polyacrylamide gel electrophoresis and alpha- and beta- naphthyl acetates as substrates. Fifty-one esterase bands were detected and classified as 31 alpha-esterases, 18 beta-esterases and two alpha/beta esterases. On the basis of the inhibition patterns using Malathion and eserine sulfate, 34 bands were classified as carboxylesterases, 14 as acethylesterases and three as cholinesterases. Ten gene loci were tentatively established on the basis of data on band position in the gel, substrate preference and inhibition pattern. Twenty bands were species-specific, the remaining being shared by species from the same or different subgroups. Bands detected exclusively in males and bands with a different frequency or degree of expression between sexes were also detected. In the gels prepared for analysis of gene expression in the body parts (head, thorax and abdomen), the degree of expression of the beta-esterases was higher in the thorax, while the alpha-esterases were expressed predominantly in the abdomen and thorax. A global view of the data available at present on the esterases of the species from the saltans group and their degree of polymorphism are presented, as well as the possibility of using some beta-esterases, because of their characteristics in the gels, as markers for species identification. PMID- 19301128 TI - Heavy metals in potable groundwater of mining-affected river catchments, northwestern Romania. AB - Groundwater, accessed using wells and municipal springs, represents the major source of potable water for the human population outside of major urban areas in northwestern Romania, a region with a long history of metal mining and metallurgy. The magnitude and spatial distribution of metal contamination in private-supply groundwater was investigated in four mining-affected river catchments in Maramures and Satu Mare Counties through the collection of 144 groundwater samples. Bedrock geology, pH and Eh were found to be important controls on the solubility of metals in groundwater. Peak metal concentrations were found to occur in the Lapus catchment, where metal levels exceed Dutch target and intervention values in up to 49% and 14% of samples, respectively. A 700 m wide corridor in the Lapus catchment on either side of the main river channel was identified in which peak Cd (31 MUg l(-1)), Cu (50 MUg l(-1)), Pb (50 MUg l(-1)) and Zn (3,000 MUg l(-1)) concentrations were found to occur. Given the generally similar bedrock geologies, lower metal levels in other catchments are believed to reflect differences in the magnitude of metal loading to the local environment from both metal mining and other industrial and municipal sources. Sampling of groundwater in northwestern Romania has indicated areas of potential concern for human health, where heavy metal concentrations exceed accepted environmental quality guidelines. The presence of elevated metal levels in groundwater also has implications for the implementation of the EU Water Framework Directive (WFD) and achieving 'good' status for groundwater in this part of the Danube River Basin District (RBD). PMID- 19301129 TI - Degradation of endocrine disrupting chemicals by genetic transformants in Irpex lacteus with an inducible laccase gene of Phlebia tremellosa. AB - Irpex lacteus was genetically transformed using an laccase expression vector to get increased laccase producing strains. Stable integration of the vector was confirmed by PCR using the vector-specific primers, and the transformants showed increased laccase activities. When the transformants were grown with several endocrine disrupting chemicals, laccase activity of each transformant was induced up to six times higher than that of the wild type. They showed increased degrading activities against EDCs as well as increased removal rates of estrogenic activities generated by the EDCs than the wild type, and the laccase expression was increased during the degradations of the EDCs. PMID- 19301130 TI - Biodegradation of diesel fuel by a microbial consortium in the presence of 1 alkoxymethyl-2-methyl-5-hydroxypyridinium chloride homologues. AB - Fast development of ionic liquids as gaining more and more attention valuable chemicals will undoubtedly lead to environmental pollution. New formulations and application of ionic liquids may result in contamination in the presence of hydrophobic compounds, such as petroleum mixtures. We hypothesize that in the presence of diesel fuel low-water-soluble ionic liquids may become more toxic to hydrocarbon-degrading microorganisms. In this study the influence of 1 alkoxymethyl-2-methyl-5-hydroxypyridinium chloride homologues (side-chain length from C(3) to C(18)) on biodegradation of diesel fuel by a bacterial consortium was investigated. Whereas test performed for the consortium cultivated on disodium succinate showed that toxicity of the investigated ionic liquids decreased with increase in side-chain length, only higher homologues (C(8)-C(18)) caused a decrease in diesel fuel biodegradation. As a result of exposure to toxic compounds also modification in cell surface hydrophobicity was observed (MATH). Disulphine blue active substances method was employed to determine partitioning index of ionic liquids between water and diesel fuel phase, which varied from 1.1 to 51% for C(3) and C(18) homologues, respectively. We conclude that in the presence of hydrocarbons acting as a solvent, the increased bioavailability of hydrophobic homologues is responsible for the decrease in biodegradation efficiency of diesel fuel. PMID- 19301131 TI - Neural network detects the effects of p-CPA pre-treatment on brain electrophysiology in a rat model of focal brain injury. AB - OBJECTIVE: To examine the performance of Artificial Neural Network (ANN) in evaluation of the effects of pretreatment of para-Chlorophenylalanine (p-CPA), a serotonin blocker, in experimental brain injury. METHODS: Continuous 4 h digital electroencephalogram (EEG) recordings from male Charles Foster rats and its power spectrum analysis by using fast Fourier transform (FFT) were performed in two experimental (i) drug untreated injury group; (ii) p-CPA pretreated injury group as well as a control group. The EEG power spectrum data were tested by ANN containing 60 nodes in input layer, weighted from the digital values of power spectrum from 0 to 30 Hz, 18 nodes in hidden layer and an output node. The effects of injury and of the drug pretreatment were confirmed with the help of calculation of edematous swelling in the brain. RESULTS: The changes in EEG spectral patterns were compared with the ANN and the accuracy was determined in terms of percent (%). Overall performance of the network was found the best in control group (97.9%) in comparison to p-CPA untreated injury group (96.3%) and p CPA pretreated injury group (71.9%). The decrease in accuracy in p-CPA pretreated injury group of subjects have occurred due to increase in misclassified patterns due to faster recovery in brain cortical potentials. CONCLUSION: EEG spectrum analysis with ANN was found successful in identifying the changes due to brain swelling as well as the effect of pretreatment of p-CPA in focal brain injury condition. Thus, the training and testing of ANN with EEG power spectra can be used as an effective diagnostic tool for early prediction and monitoring of brain injury as well as the effects of drugs in this condition. PMID- 19301132 TI - Improved accuracy of cardiac output estimation by the partial CO2 rebreathing method. AB - OBJECTIVE: This study investigated the accuracy of the NICO monitor equipped with the newer software. Additionally, the effects of the increased dead space produced by the NICO monitor on ventilatory settings were investigated. METHODS: Forty-two patients undergoing elective aortic reconstruction participated in this prospective, observational study at a university hospital. Cardiac output was continuously monitored using both the NICO monitor and continuous cardiac output (CCO) measured by a pulmonary artery catheter. A NICO monitor equipped with ver. 4.2 software was used for the first 21 patients while a NICO monitor equipped with ver. 5.0 software was used for the rest of the patients. Cardiac output measured by bolus thermodilution (BCO) at 30 min intervals was used as a reference. RESULTS: The bias +/- precision of the NICO monitor was 0.18 +/- 0.88 l/min with ver. 4.2 software (n = 182) and 0.18 +/- 0.83 l/min with 5.0 software (n = 194). The accuracy of the NICO monitor is comparable to CCO, whose bias +/- precision against BCO is 0.19 +/- 0.81 l/min (n = 376). At the same level of CO(2) production and minute ventilation, PaCO(2) was lower in the patients monitored by NICO with ver. 5.0 software than patients with ver. 4.2 software. CONCLUSIONS: This study demonstrated the improved performance of the NICO monitor with updated software. The performance of the NICO monitor with ver. 4.2 or later software is similar to CCO. However, the cardiac output measurement did not fulfill the criteria of interchangeability to the cardiac output measurement by bolus thermodilution. Updates to ver. 5.0 attenuated the effects of rebreathing introduced by the NICO monitor without compromising the accuracy of the cardiac output measurement. PMID- 19301133 TI - Flow-through versus sidestream capnometry for detection of end tidal carbon dioxide in the sedated patient. AB - BACKGROUND: End tidal carbon dioxide (ETCO(2)) in non-intubated patients can be monitored using either sidestream or flow-through capnometry [Yamamori et al., J Clin Monit Comput 22(3):209-220, 2008]. The hypothesis of this validation study is that, flow-through capnometry will yield a more accurate estimate of ETCO(2) than sidestream capnometry when evaluated in a bench study during low tidal volumes and high oxygen administration via nasal cannula. Secondarily, when ETCO(2) from each is compared to arterial CO(2) (PaCO(2)) during a study in which healthy, non-intubated volunteers are tested under normocapnic, hypocapnic and hypercapnic conditions, the flow-through capnometer will resemble PaCO(2) more closely than the sidestream capnometer. This will be especially true during periods of lower minute ventilation and high oxygen flow rates via mask in non intubated, remifentanil sedated, healthy volunteers whose physiologic deadspace is small. METHODS: The performance of a flow-through (cap-ONE, Nihon Kohden, Tokyo, Japan) and a sidestream (Microcap Smart CapnoLine Plus, Oridion Inc., Needham, MA) capnometer were compared in a bench study and a volunteer trial. A bench study evaluated ETCO(2) accuracy using waveforms generated via mechanical lungs during low tidal volumes and high oxygen flow rates. A volunteer study compared the ETCO(2) for each capnometer against PaCO(2) during sedation in which 8 l O(2) was delivered via mask rather than the nasal cannula. RESULTS: In the bench study, the flow-through capnometer gave slightly higher values of ETCO(2) during high-flow oxygen and no discernable differences during variable tidal volumes. Bland and Altman plots comparing ETCO(2) to PaCO(2) showed essentially equal performance between the two capnometers in the volunteers. CONCLUSIONS: Within a wide limit of agreement between the volunteer and bench study, flow through and sidestream capnometry performed equally well during bench testing and in non-intubated, sedated patients. PMID- 19301134 TI - C-reactive protein and ovarian cancer: a prospective study nested in three cohorts (Sweden, USA, Italy). AB - OBJECTIVES: Inflammatory processes may influence the risk of epithelial ovarian cancer, but available epidemiological evidence is limited and indirect. Circulating C-reactive protein (CRP), a sensitive marker of inflammation, may serve as a direct biological marker of an underlying association. METHODS: The association between ovarian cancer risk and pre-diagnostic circulating CRP was tested in a case-control study nested within three prospective cohorts from Sweden, USA, and Italy. The study included 237 cases and 427 individually matched controls. CRP was measured in stored blood samples by high-sensitivity immunoturbidimetric assay. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by conditional logistic regression. RESULTS: Overall, CRP was not related to risk of ovarian cancer. However, a marked increase in risk was observed for CRP concentrations >10 mg/l: OR (95% CI) 4.4 (1.8-10.9), which remained significant after limiting analyses to cases diagnosed more than two or five years after blood donation (OR 3.0 (1.2-8.0) and 3.6 (1.0-13.2), respectively). Risk of mucinous tumors increased with high CRP, but the number of cases in this analysis was small. CONCLUSION: Study results offer additional support to the concept that chronic inflammation plays a role in epithelial ovarian cancer. PMID- 19301135 TI - MassBuilt: effectiveness of an apprenticeship site-based smoking cessation intervention for unionized building trades workers. AB - OBJECTIVE: Blue-collar workers are difficult to reach and less likely to successfully quit smoking. The objective of this study was to test a training site-based smoking cessation intervention. METHODS: This study is a randomized controlled trial of a smoking cessation intervention that integrated occupational health concerns and was delivered in collaboration with unions to apprentices at 10 sites (n = 1,213). We evaluated smoking cessation at 1 and 6 months post intervention. RESULTS: The baseline prevalence of smoking was 41%. We observed significantly higher quit rates in the intervention versus control group (26% vs. 16.8%; p = 0.014) 1 month after the intervention. However, the effects diminished over time so that the difference in quit rate was not significant at 6 month post intervention (9% vs. 7.2%; p = 0.48). Intervention group members nevertheless reported a significant decrease in smoking intensity (OR = 3.13; 95% CI: 1.55 6.31) at 6 months post-intervention, compared to controls. CONCLUSION: The study demonstrates the feasibility of delivering an intervention through union apprentice programs. Furthermore, the notably better 1-month quit rate results among intervention members and the greater decrease in smoking intensity among intervention members who continued to smoke underscore the need to develop strategies to help reduce relapse among blue-collar workers who quit smoking. PMID- 19301136 TI - The winged-helix transcription factor JUMU is a haplo-suppressor/triplo-enhancer of PEV in various tissues but exhibits reverse PEV effects in the brain of Drosophila melanogaster. AB - Gene expression goes along with changes in chromatin structure and is regulated by chromatin-modifying factors. If genes are transposed from their euchromatic position to the vicinity of heterochromatin, their expression can underly a position effect variegation (PEV). In Drosophila melanogaster a few genes are known that function in a gene dose-dependent manner as haplo-suppressors and triplo-enhancers of PEV or vice versa. The gene jumeaux (jumu) encodes a winged helix transcription factor of multiple regulatory functions. A novel PEV test system for Drosophila melanogaster reveals that JUMU behaves as a haplo suppressor/triplo-enhancer in different larval and adult tissues, but surprisingly behaves in the reverse manner as a haplo-enhancer/triplo-suppressor in larval and adult brains. Like jumu, the Su(var)3-9 gene also behaves as a haplo-suppressor/triplo-enhancer, but in our test system does not show any PEV effect in the brains. PMID- 19301137 TI - Strategic noise map of a major road carried out with two environmental prediction software packages. AB - The main objective of this study is to analyze the differences found in the results of noise mapping using two of the most popular software techniques for the prediction of environmental noise. The location selected to conduct the comparative study is an area encompassed by the ring road that surrounds the city of Pamplona and on a grid, with a total of 6 x 10(5) points, approximately. In fact, and as the Environmental Noise Directive points out, it is a major road designated by a Member State (Spain). Configuration of the calculation parameters (discretization of the sources, ground absorption, reflection order, etc.) was as equivalent as possible as far as programs allow. In spite of that, a great number of differences appear in the findings. Although in 95.5% of the points the difference in the noise level calculated from the two programs was less than 3 dB, this general statistic result concealed some great differences. These are due to the various algorithms that programs implement to evaluate noise levels. Most differences pertain to highly screened receivers or remote ones. In the former, the algorithm of visibility is the main cause of such differences. In the latter, differences are mainly brought about by a different implementation of the propagation under homogeneous and favorable atmospheric conditions from both software systems. PMID- 19301138 TI - Evaluating pollution potential of leachate from landfill site, from the Pune metropolitan city and its impact on shallow basaltic aquifers. AB - Leachate produced by municipal solid waste dumping site near the metropolitan city of Pune, India was examined for its pollution potential and impact on surrounding shallow basaltic aquifers. Twenty-eight physico-chemical parameters during post- and pre-monsoon seasons (Nov 2006 and May 2007) were determined to assess the seasonal variation in the leachate pollution index (LPI) as well as in the groundwater quality. The leachate demonstrated higher LPI value during pre monsoon, comparable to those at other metropolises outside India. Potentially toxic leachates derived from the dumping site have largely influenced the adjoining basaltic aquifers through two different modes of transport. Despite high contents of heavy metals (Al, Cd, Cr, Cu, Co, Fe, Mn, Ni, Pb and Zn) in the leachate, the aquifers in the close proximity of landfill site are least polluted by metallic contaminants possibly due to redox controls. Various geoenvironmental features governing the dispersal of leachate contaminants in the basaltic aquifers under semi-arid climatic regime have been identified and discussed. Although a few remedial measures have been suggested to mitigate the impact of leachate percolation and dispersion, the present study demands for a proper solid waste management in metropolitan cities. PMID- 19301139 TI - An efficient protocol for the synthesis of 2-amino-4,6-diphenylpyridine-3 carbonitrile using ionic liquid ethylammonium nitrate. AB - Chalcones on condensation with malononitrile and ammonium acetate in the presence of ionic liquid ethylammonium nitrate affords the corresponding 2-amino-4, 6 diphenylpyridine-3-carbonitrile in excellent yield. The ionic liquid is recycled and reused several times. PMID- 19301140 TI - Hematologic abnormalities following renal transplantation. AB - Recipients of renal allografts are surviving longer and, consequently, may experience a variety of complications related not only to the transplanted kidney, but also to the hematopoietic system. Common hematologic complications in the renal transplant patient include abnormalities of one cell line, such as post transplantation erythrocytosis or anemia, that are often treatable with simple measures. Conversely, pathologies involving the leukocyte and platelet population often exist in the context of pancytopenia, which may be a manifestation of systemic infection (e.g., cytomegalovirus, human herpesvirus 8) or malignancy (post-transplantation lymphoproliferative disorders). Uncommon, but life threatening, processes complicating renal transplantation include hepatosplenic gammadelta T-cell lymphoma and viral-induced hemophagocytic syndrome, both of which are associated with severe pancytopenia and, often, death. Since this patient population is often managed in a multidisciplinary fashion by nephrologists, infection specialists, transplant surgeons, hematologists, and internal medicine physicians, a succinct review of this topic is warranted. PMID- 19301141 TI - Self-propagating beta-sheet polypeptide structures as prebiotic informational molecular entities: the amyloid world. AB - The idea is advanced that under the extreme earth conditions for ~3.9 billions years ago, protein-based beta-sheet molecular structures were the first self propagating and information-processing biomolecules that evolved. The amyloid structure of these aggregates provided an effective protection against the harsh conditions known to decompose both polyribonucleotides and natively folded polypeptides. In the prebiotic amyloid world, both the replicative and informational functions were carried out by structurally stable beta-sheet protein aggregates in a prion-like mode involving templated self-propagation and storage of information in the beta-sheet conformation. In this amyloid (protein) first, hybrid replication-metabolism view, the synthesis of RNA, and the evolvement of an RNA-protein world, were later, but necessary events for further biomolecular evolution to occur. I further argue that in our contemporary DNA<- >RNA-->protein world, the primordial beta-conformation-based information system is preserved in the form of a cytoplasmic epigenetic memory. PMID- 19301142 TI - Evaluation of coupled perturbed and density functional methods of computing the parity-violating energy difference between enantiomers. AB - We present new coupled-perturbed Hartree-Fock (CPHF) and density functional theory (DFT) computations of the parity-violating energy difference (PVED) between enantiomers for H(2)O(2) and H(2)S(2). Our DFT PVED computations are the first for H(2)S(2) and the first with the new HCTH and OLYP functionals. Like other "second generation" PVED computations, our results are an order of magnitude larger than the original "first generation" uncoupled-perturbed Hartree Fock computations of Mason and Tranter. We offer an explanation for the dramatically larger size in terms of cancellation of contributions of opposing signs, which also explains the basis set sensitivity of the PVED, and its conformational hypersensitivity (addressed in the following paper). This paper also serves as a review of the different types of "second generation" PVED computations: we set our work in context, comparing our results with those of four other groups, and noting the good agreement between results obtained by very different methods. DFT PVEDs tend to be somewhat inflated compared to the CPHF values, but this is not a problem when only sign and order of magnitude are required. Our results with the new OLYP functional are less inflated than those with other functionals, and OLYP is also more efficient computationally. We therefore conclude that DFT computation offers a promising approach for low-cost extension to larger biosystems, especially polymers. The following two papers extend to terrestrial and extra-terrestrial amino acids respectively, and later work will extend to polymers. PMID- 19301143 TI - Molecular characterization and expression analysis of MHC class II alpha and beta genes in large yellow croaker (Pseudosciaena crocea). AB - MHC class II molecules play an important role in the activation of CD4(+) T cells, which are the central orchestrating cells of an immune response. Here, we report the cloning of MHC class II alpha and beta cDNAs from large yellow croaker (Pscr-DAAs and Pscr-DAB) by expressed sequence tags analysis and RACE-PCR techniques. Three different class II alpha and two class II beta sequences were obtained from spleens of two individual fish. Each of the three class II alpha sequences encodes a polypeptide of 239 amino acids while the two class II beta cDNA sequences encode for a protein of 249 aa. All the characteristic features of MHC class II chain structure could be identified in the deduced proteins of three class II alpha and two class II beta sequences, including the leader peptide, alpha1/beta1 and alpha2/beta2 domains, connecting peptide and transmembrane and cytoplasmic regions, as well as conserved cysteines and N-glycosylation site. RT PCR analysis showed that MHC class II alpha and beta mRNAs were broadly expressed in various tissues examined, although at different levels. Upon stimulation with inactivated trivalent bacterial vaccine or polyinosinic polycytidylic acid (poly(I:C)), the expression levels of both alpha and beta genes were obviously up regulated in intestine, kidney and spleen. Real-time PCR analysis demonstrated that the expression levels of class II alpha and beta were quickly up-regulated in spleen, kidney, and intestine at 12 h after induction with poly(I:C), while their expression levels significantly increased at 48 h upon immunization with bacterial vaccine, indicating that the up-regulation of both class II alpha and beta expression was induced by bacterial vaccine or poly(I:C) at the early phase of induction, and that class II alpha and beta transcripts were quicker up regulated by poly I:C than by bacterial vaccine. PMID- 19301144 TI - Transgenic RNAi-mediated reduction of LZP3 in Lagurus lagurus oocytes results in decreased fertilization ability in IVF. AB - Structural differences in oligosaccharides on mammalian zona pellucida 3(ZP3) from different species may determine whether or not spermatozoa being able to bind to ZP. We reported here that by microinjecting the siRNA interference recombinant construct pGenesil-ZP31 encoding a Lagurus zp3 (lzp3) hairpin dsRNA of 21 bp into the inmatured oocytes of Lagurus lagurus, distributed in northern region of Xingjiang, to disturb its fertility. Results of in vitro fertilization after in vitro maturation of the immature oocytes of Lagurus lagurus showed that the fertilization rate of the transgenic oocytes carried pGenesil-ZP31 was decreased greatly (2.82%) compared to the oocytes carried pGenesil-HK (15.71%), suggesting that the transgenic RNAi-mediated silencing of lzp3 in Lagurus lagurus oocytes results in decreased fertilization ability. These results proved that LZP3 of Lagurus lagurus, like other mammalians, is essential for the recognition between oocyte and spermatozoa. PMID- 19301145 TI - Reinterpreting the 'quickening' perspective in the abortion debate. AB - Personhood constitutes the pivotal point in the abortion debate. There exists a diversity of views as to when foetal personhood actually starts-from conception and implantation to viability and even birth. One perspective that has lost support for decades is that of quickening, a stance associated with Lord Ellenborough's 1803 Act. This paper attempts to put quickening back into the limelight, albeit through a new interpretation. After discussing its philosophy and underpinning rationale, I will assess a number of arguments that have been directed against quickening as a viable point of distinction. I conclude by suggesting that according to modern proponents of quickening proponents, rational soul ensoulment begins after a certain degree of cerebral cortical formation has been realized, thus marking foetal volition, which promotes foetal interests, for the first time. PMID- 19301146 TI - ATP release from non-excitable cells. AB - All cells release nucleotides and are in one way or another involved in local autocrine and paracrine regulation of organ function via stimulation of purinergic receptors. Significant technical advances have been made in recent years to quantify more precisely resting and stimulated adenosine triphosphate (ATP) concentrations in close proximity to the plasma membrane. These technical advances are reviewed here. However, the mechanisms by which cells release ATP continue to be enigmatic. The current state of knowledge on different suggested mechanisms is also reviewed. Current evidence suggests that two separate regulated modes of ATP release co-exist in non-excitable cells: (1) a conductive pore which in several systems has been found to be the channel pannexin 1 and (2) vesicular release. Modes of stimulation of ATP release are reviewed and indicate that both subtle mechanical stimulation and agonist-triggered release play pivotal roles. The mechano-sensor for ATP release is not yet defined. PMID- 19301147 TI - A psycholinguistic model of natural language parsing implemented in simulated neurons. AB - A natural language parser implemented entirely in simulated neurons is described. It produces a semantic representation based on frames. It parses solely using simulated fatiguing Leaky Integrate and Fire neurons, that are a relatively accurate biological model that is simulated efficiently. The model works on discrete cycles that simulate 10 ms of biological time, so the parser has a simple mapping to psychological parsing time. Comparisons to human parsing studies show that the parser closely approximates this data. The parser makes use of Cell Assemblies and the semantics of lexical items is represented by overlapping hierarchical Cell Assemblies so that semantically related items share neurons. This semantic encoding is used to resolve prepositional phrase attachment ambiguities encountered during parsing. Consequently, the parser provides a neurally-based cognitive model of parsing. PMID- 19301148 TI - The role of competitive learning in the generation of DG fields from EC inputs. AB - We follow up on a suggestion by Rolls and co-workers, that the effects of competitive learning should be assessed on the shape and number of spatial fields that dentate gyrus (DG) granule cells may form when receiving input from medial entorhinal cortex (mEC) grid units. We consider a simple non-dynamical model where DG units are described by a threshold-linear transfer function, and receive feedforward inputs from 1,000 mEC model grid units of various spacing, orientation and spatial phase. Feedforward weights are updated according to a Hebbian rule as the virtual rodent follows a long simulated trajectory through a single environment. Dentate activity is constrained to be very sparse. We find that indeed competitive Hebbian learning tends to result in a few active DG units with a single place field each, rounded in shape and made larger by iterative weight changes. These effects are more pronounced when produced with thousands of DG units and inputs per DG unit, which the realistic system has available, than with fewer units and inputs, in which case several DG units persists with multiple fields. The emergence of single-field units with learning is in contrast, however, to recent data indicating that most active DG units do have multiple fields. We show how multiple irregularly arranged fields can be produced by the addition of non-space selective lateral entorhinal cortex (lEC) units, which are modelled as simply providing an additional effective input specific to each DG unit. The mean number of such multiple DG fields is enhanced, in particular, when lEC and mEC inputs have overall similar variance across DG units. Finally, we show that in a restricted environment the mean size of the fields is unaltered, while their mean number is scaled down with the area of the environment. PMID- 19301149 TI - The ambiguous role of the Na+-H+ exchanger isoform 1 (NHE1) in leptin-induced oxidative stress in human monocytes. AB - Leptin, a 16-kDa cytokine produced mainly by the adipose tissue, is known to increase energy expenditure while at the same time lowering food intake by acting directly on the hypothalamus. ObRb, the leptin receptor mostly involved in intracellular signaling, is expressed in a wide range of tissues, thus allowing leptin to affect a much broader diversity of biological processes. High concentrations of leptin are encountered in patients with hyperleptinemia, a condition which very often accompanies obesity and which is a direct result of leptin resistance. In the present study, moderate and high concentrations of leptin (16 and 160 ng/ml) were mostly utilized in order to investigate the role of this cytokine in oxidative stress levels in human monocytes. Leptin was found to increase oxidative species production as measured with 2',7' dichlorodihydrofluorescein diacetate (general marker of oxidative species, but not O2-*) and dihydroethidium (marker of O2-*). Surprisingly, it also augmented superoxide dismutase activity. Inhibition of the Na+-H+ exchanger isoform 1 (NHE1) also inhibited leptin-induced superoxide anion production but at the same time amplified leptin-induced production of other oxidative species. Signaling proteins such as phosphoinositide 3 kinase and conventional isoforms of protein kinase C (alpha-, beta(i)-, beta(ii)-), as well as NADPH oxidase, also participated in leptin signaling. Finally, leptin was found to increase glutathionylation levels of NHE1-bound heat shock protein 70 kDa (Hsp70) but not Hsp70 binding to NHE1. PMID- 19301150 TI - Hairy cell leukemia-related disorders consistently show low CD27 expression. AB - In Japan, typical hairy cell leukemia (HCL) is rare, and HCL-Japanese variant (HCL-JV) is more common. Hairy B-cell lymphoproliferative disorder (HBLD) is another unusual disorder of polyclonal B-lymphocytosis of hairy cell appearance. In the present study, we analyzed the clinical features of 3 patients with HCL, 3 with HCL-JV, and 3 with HBLD. All HBLD patients had the DRB1*04 allele. As compared with other B-cell lymphoproliferative disorders, CD27 expression on B cells was significantly lower in all patients, ranging from 0.3% to 23.4%. Our results suggest that low CD27 expression may be a distinct feature of these HCL related disorders. PMID- 19301151 TI - Immunohistochemical prognostic markers in intracranial ependymomas: systematic review and meta-analysis. AB - Distinction between grade II ependymomas and anaplastic ependymomas based on histopathological examination solely is problematic and, therefore, the management of intracranial ependymomas remains controversial. The aim of this study was to conduct a systematic review (SR) and meta-analysis (MA) of data published on immunohistochemical prognostic markers (IPM) in intracranial ependymomas (IE), and to establish an evidence-based perspective on their clinical value. Following the extensive search based on a strictly defined group of key words, 30 studies reporting results on IPM in IE were identified. Due to a pronounced inter-study heterogeneity, only 14 publications fulfilled the criteria for inclusion into SR. From the total of 67 immunohistochemical markers, 18 were found to correlate with prognosis. However, owing to inadequate data publishing, MA could be performed only with data on proliferation marker MIB-1 (Ki-67) from 5 publications, including 337 patients: The pooled hazard ratio for overall survival was 3.16 (95% confidence interval = 1.96-5.09; p < 0.001) implicating that patients suffering from tumors with higher immunohistochemical expression of MIB-1 had a significantly worse outcome. Marked inter-study heterogeneity and incomplete data publishing in primary studies significantly limited extent of the SR, and the possibility of performing MA. Although the prognostic impact of MIB-1 immunoexpression in IE could be confirmed, there remains lack of further reliable IPM that could be used in routine diagnosis. We encourage to search for new, useful markers, as well as to standardize lab-techniques and data interpretation algorithms across laboratories in order to increase data compatibility. PMID- 19301152 TI - Growth hormone production and action in N1E-115 neuroblastoma cells. AB - Neuroblastoma cells are undifferentiated cells derived from the neural crest and are commonly used as models for studying neural function. Mouse N1E-115 neuroblastoma cells are derived from cancerous tissue and provide a model for studying the oncogenesis of neural cells. As growth hormone (GH) has been implicated as an autocrine or paracrine involved in neural regulation and in the induction or progression of cancer, the possibility that N1E-115 cells are sites of GH production and GH action was assessed. Using RT-PCR, cultured N1E-115 cells were found to express the mouse GH and GH receptor (GHR) genes. Immunocytochemistry demonstrated that both of the translated proteins (GH and its receptor) were abundantly present in the cytoplasm of these cells and their co localization was established by confocal cytochemistry. GH action in these cells was determined in cells cultured for 72 h in the presence or absence of 10(-6) M or 10(-9) M mouse GH, which induced neurite sprouting and increased axon growth. In summary, the expression of GH and its receptor in GH responsive tumor-derived N1E-115 neuroblastoma cells suggests they provide a useful experimental model to assess GH actions in neural function or neural oncogenesis. PMID- 19301153 TI - When percutaneous dilation tracheotomy may be hazardous: abnormal course of the brachiocephalic trunk. AB - INTRODUCTION: Percutaneous dilatational tracheotomy (PDT) is becoming increasingly popular in present day critical care medicine. In contrast to the surgical approach, PDT involves a blind puncture and dilation of the pretracheal space, which may predispose to dangerous complications in the case of anatomical, in particular vascular, anomalies. METHODS AND RESULTS: We report on two patients, in whom an abnormal pulsation was detected when the infracricoid region was palpated in preparation for PDT. An immediately performed ultrasound scan revealed an arterial blood vessel in front of the upper part of the trachea. A subsequent CT-angiography showed an anomalous course of the brachiocephalic trunk. While too dangerous for PDT, the local department of cranio-maxillofacial surgery was consulted for surgical tracheotomy. CONCLUSION: To avoid hazardous bleeding complications in PDT we recommend at least an ultrasound scan in case of an abnormal pulsation and an enlarged thyroid gland. PMID- 19301154 TI - Ectopic growth hormone-releasing hormone secretion by a neuroendocrine tumor causing acromegaly: long-term follow-up results. AB - Ectopic secretion of growth hormone-releasing-hormone (GHRH) is a rare cause of acromegaly-representing less than 1% of patients. A 25-year-old woman was admitted to the hospital with acromegaly and a 6 x 6 cm infrahepatic mass. Sellar magnetic resonance imaging indicated diffuse pituitary enlargement consistent with hyperplasia. The infrahepatic mass was resected, and the histopathological diagnosis was a well-differentiated invasive neuroendocrine carcinoma of the duodenum with metastases to local lymph nodes. The tumor cells contained cytoplasmic immunoreactivity for GHRH. Because increased IGF-1 concentrations persisted after the operation, the patient was treated with octreotide long acting repeatable (LAR) injections of 20 mg/month. Growth hormone and IGF-1 levels normalized. After 6 years of surveillance, a left paraaortic mass was detected by uptake of indium 111 octreotide. Surgical exploration revealed metastatic neuroendocrine carcinoma in a 2.5-cm lymph node. Postoperatively, the IGF-1 concentration was mildly elevated. Octreotide LAR therapy is being continued at 10 mg/month. This case suggests that octreotide treatment may have a beneficial effect on disease course and can be maintained for as long as 7 years in a patient with acromegaly due to a GHRH-secreting neuroendocrine carcinoma. PMID- 19301155 TI - Phylogenetic analysis and in silico characterization of the GARS-AIRS-GART gene which codes for a tri-functional enzyme protein involved in de novo purine biosynthesis. AB - Human GARS-AIRS-GART encodes a fused tri-functional enzyme protein involved in de novo purine biosynthesis, aberrant function being implicated in Down syndrome and Leukemia. We performed phylogenetic analysis to discern evolutionary relationships and in silico characterization to identify elements potentially important for gene regulation. We report that murine, bovine and chimpanzee sequences are the nearest neighbors of human GARS-AIRS-GART and that endo duplication of the AIRS protein is restricted to insect orthologs. Convergent evolution of mono-functional bacterial orthologs to bi-functional, partly fused, yeast orthologs is observed from the rooted-NJ tree topology that bears bootstrap values exceeding 9000 in majority of the nodes. Sequence alignments reveal that introns 11-15 of human GARS-AIRS-GART are conserved among vertebrates. An inverse correlation is observed between intron size and intron density without bias for intron position. The generation time of organisms is independent of intron density. Human, bovine and murine sequences possess similar GC content with CpG islands in promoter regions. The long isoforms of cow and chicken transcripts and short isoforms of human, bovine and murine mRNA form energetically stable stem like structures in the 3'-UTR and may regulate translational stability of GARS AIRS-GART transcripts. Glycine-rich loops important for enzyme structure and ATP , folate-binding residues are partially conserved. PMID- 19301156 TI - Lambda chops: creation of site-directed mutants in insertable fragments utilizing Gateway technology. AB - We describe a method to produce site-directed mutations anywhere within cDNA by assembling mutagenized PCR fragments in proper orientation using lambda integration in an extension of Gateway technology to yield a full-length mutated gene. This process exploits the directionality of lambda insertion sequences ensuring integration and directionality of PCR product into a cloning vector. The process requires only two sequential integration steps to yield a mutagenized expression vector. Mutagenized vasodilator associated phosphoprotein (VASP) was produced by generating two PCR fragments representing the upstream and downstream portions of the gene, substituting alanine or glutamate residues for VASP serine239. The upstream PCR was engineered with attB1 lambda integration sequences at the 5' region and attB2 at the 3' region of the downstream fragment to ensure correct orientation. The desired mutation was encoded by the forward primer of fragment 2. The reverse primer of the fragment 1 was phosphorylated for subsequent ligation. Vent polymerase provided sequence accuracy and blunt-ended product. The first integration into a donor vector, catalyzed by BP Clonase II created a linear product circularized by blunt end ligation, yielding hundreds of entry vectors containing the mutagenized VASP. A second integration into destination vector yielded plasmid expressing mutant VASP upon transfection. PMID- 19301157 TI - The mTOR pathway is associated with the poor prognosis of human hepatocellular carcinoma. AB - OBJECTIVE: The mammalian target of rapamycin (mTOR) pathway, an important regulator of multiple cellular functions including proliferation, differentiation, tumorigenesis, and apoptosis, is up-regulated in many cancers. It has achieved considerable importance. This study was conducted to determine the status of the mTOR pathway in human hepatocellular carcinoma (HCC) and to investigate its relationship with the prognosis of HCC. METHODS: PTEN, pAkt, p27, and pS6 expression in cryo-sections gathered from 528 cases with HCC by the method of immunohistochemistry. Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of HCC. RESULTS: The mTOR pathway was more significantly altered in high-grade tumors, and tumors with poor prognostic features. Especially, pAkt and cytoplasmic p27 expression showed the strongest associations with pathological parameters of HCC. Statistical analysis showed that HCC patients expressing pAkt, PTEN, cytoplasmic p27, and pS6 have different overall survival rates relative to those not expressing these proteins. Cox multi factor analysis showed that tumor differentiation (P = 0.006), vascular invasion (P = 0.028), TNM stage (P = 0.005), pAkt (P = 0.021), PTEN (P = 0.003), p27 (P = 0.018) and pS6 (P = 0.002) were independent prognosis factors for HCC. CONCLUSION: Expression of the mTOR pathway components, which are related with the transferability and invasive capacity of HCC cells, may be used as prognostic indicators in HCC. PMID- 19301158 TI - Primary colorectal lymphoma. AB - This study aimed at investigating the incidence, presentation, patient and tumor characteristics, treatment, and outcome of primary colorectal lymphomas (PCL) at a tertiary care center in Lebanon over a 25-year period. The Dawson's criteria were used for selection of eligible cases. The medical records were reviewed for demographic variables, the presence of risk factors, presenting signs and symptoms, method of diagnosis, histologic type, type of therapy, and condition at last follow-up. Nine cases of PCL were identified (12.7% of gastrointestinal lymphomas and 0.1% of colorectal malignancies). The mean age at presentation was 44.2 years with male predominance noted. Abdominal pain was the most common presentation (77.8%). Colonoscopy was performed for eight patients with non specific gross tumor characteristics. Three patients had Burkitt's and six had diffuse large B-Cell lymphomas. The most common site of involvement was the cecum (55.6%) with all cases presenting in stage I(E). Surgery was performed for six patients followed by chemotherapy except for one, and three patients had chemotherapy only. The median survival time was 25 months and the 2-year survival time was approximated at 60%. It is concluded that PCL is a rare malignancy with well-identified disease characteristics yet controversial ideal management plan. PMID- 19301159 TI - Association between periodontal disease and ischemic heart disease among Swedish women: a cross-sectional study. AB - OBJECTIVE: The aim of this cross-sectional study was to analyze the relationship between chronic periodontitis and ischemic heart disease (IHD). MATERIAL AND METHODS: A cross-section of women aged 38 to 84 years were examined in 1992-93 (analysis based on n=1056). Medical and dental examinations were included in the analysis specifically with regard to IHD and periodontitis. Other well-known risk factors for IHD were used as covariates in multivariable statistical analysis. RESULTS: Among the dentate women in this study (n=847), 74 had IHD and 773 did not. There was no statistically significant difference between numbers of pathological gingival pockets between these groups (58.1% had one or more pathological pockets in the IHD group compared to 57.6% in the non-IHD group). Bivariate analysis of dentate individuals showed significant associations between IHD and number of missing teeth, age, body mass index, waist/hip ratio, life satisfaction, hypertension, and levels of cholesterol and triglycerides. However, in the final multivariable logistic regression model, with the exception of age, only number of teeth (<17 teeth) OR = 2.13 (CI 1.20; 3.77) was found to be significantly associated with IHD. Moreover, edentulous women had an OR of 1.94 (CI 1.05; 3.60) in relation to IHD (age-adjusted model). CONCLUSIONS: In the present study, periodontitis did not seem to have a statistically significant relationship with IHD. The number of missing teeth showed a strong association with IHD, and this may act as a proxy variable tapping an array of different risk factors and behaviors. PMID- 19301160 TI - Effects of improvement in periodontal inflammation by toothbrushing on serum lipopolysaccharide concentration and liver injury in rats. AB - OBJECTIVE: Periodontitis increases the serum lipopolysaccharide level, contributing to liver injury. Toothbrushing improves periodontitis and may also affect serum lipopolysaccharide concentration and periodontitis-induced liver injury. The purpose of the present study was to examine whether the improvement in periodontal inflammation by toothbrushing clinically affects the serum lipopolysaccharide level and hepatic pathological changes in rat periodontitis. MATERIAL AND METHODS: Thirty male Wistar rats were divided into 5 groups, 2 groups receiving topical application of pyrogen-free water to the gingival sulcus for 4 or 8 weeks. The next 2 groups received topical application of lipopolysaccharide and proteases for 4 or 8 weeks. The last group received topical application of lipopolysaccharide and proteases for 8 weeks, and the palatal gingiva was brushed with a powered toothbrush once a day for 4 weeks prior to the end of the experimental period. RESULTS: Topical application of lipopolysaccharide and proteases induced not only periodontal inflammation but also an elevation in the serum lipopolysaccharide concentration, with increasing hepatic inflammation, steatosis and 8-hydroxydeoxyguanosine levels in a time dependent manner. The rats that received gingival stimulation showed decreased polymorphonuclear leukocyte infiltration and collagen loss levels in the periodontal lesions. Furthermore, this group also showed a decrease in serum lipopolysaccharide concentration and hepatic inflammation, steatosis and 8 hydroxydeoxyguanosine levels, compared with the group receiving no treatment. CONCLUSIONS: Toothbrushing promoted healing of periodontal lesions, decreased serum lipopolysaccharide concentration and suppressed liver injury in a rat periodontitis model. PMID- 19301162 TI - The limits of attention for visual perception and action in aging. AB - This study aimed to investigate whether aging results in an increased attentional blink effect in older adults as compared to young adults. A rapid serial visual presentation (RSVP) paradigm was employed in which participants were asked to identify two targets (dual-task condition) presented in rapid succession. These targets were separated by various intervals in a stream of stimuli. The performance for identifying the second target was normally diminished as compared to identification of a single-task target. Various combinations of tasks, such as two perceptual tasks or one perceptual and one action task, as well as different types of pointing action, such as pointing to a displaced target, pointing to a stationary target or pointing to a disappeared target, were manipulated in this study to see if aging may further impact these variables. The results of this study showed that in young adults, successful identification of the first target interfered with identifying the second target, as well as the initiation time (action planning) of pointing to the second target. However, identification of the first target did not interfere with pointing movement time and pointing accuracy, even when the target was displaced, which required online control of action. Conversely, for older adults, successful central identification not only interfered with identifying the second target and with the pointing initiation time, but also interfered with pointing movement time for a displaced target. This suggests that older adults seem to be unable to concurrently identify the first target and correct their already-initiated pointing movement compared to young adults. PMID- 19301163 TI - Impaired exercise capacity predicts sudden cardiac death in a low-risk population: enhanced specificity with heightened T-wave alternans. AB - AIMS: Because sudden cardiac death (SCD) is due to cardiac electrical instability, we postulated that prediction of this mode of death by exercise capacity will be enhanced by combined assessment with T-wave alternans (TWA), an index of repolarization abnormality. MATERIAL AND METHODS: The Finnish Cardiovascular Study enrolled consecutive patients (n=2,044) with a routine clinically indicated exercise test. Exercise capacity was measured in metabolic equivalents (METs) and TWA by time-domain modified moving average method. RESULTS: During 47.2+/-12.8-month follow-up (mean+/-SD) 120 patients died; 58 were cardiovascular deaths, and 29 were SCD. In multivariate analysis after adjustment for sex, age, smoking, use of beta-blockers, as well as other common coronary risk factors, the relative risk of patients whose exercise capacity was depressed (MET < 8) was 8.8 (95% CI 2.0-38.9, P=0.004) for SCD. The combination of low exercise capacity (MET < 8) and elevated TWA (> or =65 microV) yielded relative risks for SCD of 36.1 (6.3-206.0, P<0.001), for cardiovascular mortality of 21.1 (6.7-66.2, P<0.001), and for all-cause mortality of 7.8 (3.5-17.4, P<0.001) over patients with neither factor. CONCLUSIONS: Reduced exercise capacity, particularly in combination with heightened TWA, indicating enhanced cardiac electrical instability, powerfully predicts risk for SCD in patients referred for exercise testing. PMID- 19301164 TI - Substance misuse and early psychosis. AB - OBJECTIVE: This paper sought to review current knowledge about the relationship between substance misuse and early psychosis. METHODS: A literature search was conducted using Medline and restricting the search to articles after 1996. Additional articles were sourced from reference lists of relevant articles. RESULTS: There is a high prevalence of substance misuse among persons with early psychosis, with cannabis and alcohol featuring prominently. Substance misuse is associated with earlier onset and possibly more positive symptoms, although apparently not with greater cognitive impairment. Cannabis appears to confer an increased likelihood of developing schizophrenia in biologically vulnerable individuals. Amphetamines also cause psychosis which may become chronic, although specific vulnerability to this effect is less well established. Many cases of so called 'drug-induced psychosis' become diagnosed as schizophrenia in later years. Specific intervention programs report positive outcomes with regard to substance misuse and the course of psychosis. CONCLUSIONS: Substance misuse should always be assessed in this patient group, bearing in mind the potential interactive causes of psychopathology. Intervention is of value in improving outcomes. PMID- 19301165 TI - First CT findings and improvement in GOS and GOSE scores 6 and 12 months after severe traumatic brain injury. AB - PRIMARY OBJECTIVE: To analyse the association between individual initial computerized tomography (CT) scan characteristics and Glasgow Outcome Scale (GOS) and Extended Glasgow Outcome Scale (GOSE) improvement between 6 months and 1 year. METHODS AND PROCEDURES: Two hundred and twenty-four adult patients with severe traumatic brain injury and Glasgow Coma Scale (GCS) score of 8 or less who were admitted to an intensive care unit were studied. GOS and GOSE scores were obtained 6 and 12 months after injury in 203 subjects. Patients were predominantly male (84%) and median age was 35 years. MAIN OUTCOMES AND RESULTS: Traumatic Coma Data Bank (TCDB) CT classification was associated with GOS/GOSE improvement between 6 months and 1 year, with diffuse injury type I, type II and evacuated mass improving more than diffuse injury type III, type IV and non evacuated mass; for GOS 43/155 (28%) vs 3/48 (6%) (chi(2) = 9.66, p < 0.01) and for GOSE 71/155 (46%) vs 7/48 (15%) (chi(2) = 15.1, p < 0.01). CT individual abnormalities were not associated with GOS/GOSE improvement, with the exception of subarachnoid haemorrhage, which showed a negative association with GOSE improvement (chi(2) = 4.08, p < 0.05). CONCLUSIONS: TCDB CT scan classification and subarachnoid haemorrhage were associated with GOS/GOSE improvement from 6-12 months, but individual CT abnormalities were not associated. PMID- 19301166 TI - Acquired brain injury and dementia: a comparison of carer experiences. AB - OBJECTIVE: As their differential needs are unknown and to inform service planning, this study (a) examined experiences of caring for adults with acquired brain injury (ABI) and (b) compared these with carers of adults with dementia. DESIGN: Cross-sectional postal survey. ABI carer experiences were compared with those of a previously studied group of dementia carers using equivalent instruments. METHODS: Family carers (n = 222) of adults with ABI: TBI (49%), strokes (26%), brain infections (18%) and other (7%) completed validated questionnaires assessing physical dependency and psychological problems of those cared for and carers' own perceived burden, quality-of-life and mental health. RESULTS: Carer burden, quality-of-life and mental health were worse for ABI carers, but were not predicted by gender, relationship, injury type, physical dependency or cognitive problems in either ABI or dementia carers. Behavioural problems of those cared for varied between the two groups and affected carers differently. Aggressive problems significantly predicted greater burden, poor quality-of-life and mental health in ABI carers, whereas passivity/low mood significantly predicted greater burden and worse quality-of-life in dementia carers. CONCLUSIONS: This study revealed different experiences of caring for younger adults with ABI vs. older adults with dementia, thereby supporting targeted development of services to sustain families affected by these conditions. PMID- 19301167 TI - Methamphetamine use patterns among urban Black men who have sex with men. AB - The present study investigates patterns of methamphetamine, contexts of use and sources for purchasing methamphetamine in a sample of gay, bisexual and heterosexually identified Black men who have sex with men living in New York City. Participants reported using multiple substances and used, on average, nine days within the last 30 days. They spent an average of US$159 per month on methamphetamine and a median of US$398 on all substances. Frequency of monthly methamphetamine use was related to the use of powdered cocaine and alcohol use, but not to the use of other substances. Black men who have sex with men primarily used in private venues, such as at home or in the homes of friends, and reported obtaining methamphetamine from multiple sources. Men who reported exchanging sex for methamphetamine reported greater use in public venues, such as sex clubs, sex parties and circuit parties. Findings from the present study may be critical in establishing culturally-appropriate treatment modalities for Black men who have sex with men who use methamphetamine. PMID- 19301168 TI - Culture conditions shape mesenchymal stromal cell phenotype and function. PMID- 19301169 TI - Selection of CD271(+) cells and human AB serum allows a large expansion of mesenchymal stromal cells from human bone marrow. AB - BACKGROUND: Mesenchymal stromal cells (MSC) are promising candidates for cell therapy and tissue engineering and may be used to treat acute graft-versus-host disease (GvHD). However, major obstacles for their clinical use are the required cell dose and the biosafety and potential immunogenicity of fetal bovine serum (FBS), which is a crucial supplement of all media currently used for the culture of MSC. METHODS: In this study MSC were successfully expanded after selection of CD271 cells from human bone marrow (BM) mononuclear cells in medium supplemented with 10% pooled allogeneic human serum. RESULTS: We isolated MSC from 10 healthy donor BM by plastic adherence and immunomagnetic selection of the CD271(+) fraction and expanded MSC in medium supplemented with pooled human allogeneic serum and animal serum. We isolated a homogeneous multipotent population by CD271(+) selection with a proliferation rate that was higher than MSC isolated by plastic adherence, 6.8+/-1.57 compared with 2.07+/-1.40 logs. Similar to cells generated in animal serum medium, MSC from allogeneic human serum were positive for mesenchymal markers and negative for hematopoietic markers; moreover they expressed embryonic stem cell genes. A normal karyotype and differentiation capacity into adipogenic, osteogenic and chondrogenic lineages and neurosphere like structures were preserved throughout long-term culture. DISCUSSION: Expansion of MSC is both feasible and large with a CD271-selected population in medium supplemented with 10% pooled allogeneic human serum, without loss of multipotent differentiation capacity or karyotype alterations. PMID- 19301171 TI - Effects of age, gender and chronic obstructive pulmonary disease on taste acuity. AB - PRIMARY OBJECTIVE: Multifactoral interactions of age, gender and medical conditions were investigated because of lack of definitive results in previous studies. METHODS: Absolute detection and recognition were determined thresholds using a three-alternative forced-choice test by younger men (n=50) and women (n=48); and by older men (n=43) and women (n=46) for each of the four basic tastes (sweet, sour, bitter, salty). Data from 46 elderly men with chronic obstructive pulmonary disease (COPD) were included. RESULTS: Age was consistently associated with elevated recognition thresholds for each of the four tastants for men only. Gender differences were seen for absolute sweet thresholds and for sour, bitter, and sweet recognition thresholds for older participants. Men with COPD had the highest levels for absolute and recognition thresholds for each tastant. CONCLUSIONS: Gender plays a role in taste perception. COPD diminishes taste. PMID- 19301173 TI - A review of German Scedosporium prolificans cases from 1993 to 2007. AB - Scedosporium prolificans is one of the most life-threatening fungal opportunistic pathogens due to its high resistance to common systemic antifungal agents. While a close relative of Pseudallescheria boydii, S. prolificans has a more limited geographic range being primarily found in Australia, USA and Spain. Infections have also been reported from several other European countries and from Chile. Twenty patients with Scedosporium prolificans infection or colonization from August 1993 to May 2007 were retrospectively reviewed in Germany. They had all been identified at or reported to the Reference Laboratory for Pseudallescheria/Scedosporium spp. in Berlin. Twelve of 13 patients with haematological disorders and/or on immunosuppressive therapy developed a fatal invasive scedosporiosis. Colonization of the respiratory tract was reported for one patient after heart-lung-transplantation, all six patients with cystic fibrosis and one with chronic sinusitis. Molecular studies of the S. prolificans isolates confirmed that parts of the 18S, the Internal Transcribed Spacer (ITS) regions and the D1/D2 domain of the 28S region of rDNA are monomorphic. However, sequencing of parts of the translation elongation factor EF1-alpha (EF-1alpha) and the chitin synthase (CHS-1) genes revealed the presence of three and two distinct genotypes, respectively. Two informative mutations were found in EF 1alpha and a single nucleotide exchange in the CHS-1 gene. PMID- 19301174 TI - Primary cutaneous zygomycosis due to Absidia corymbifera in a patient with cutaneous T cell lymphoma. AB - A slowly enlarging arm ulcer appeared in a 61-year-old man with cutaneous T cell lymphoma. Skin biopsy revealed aseptate hyphae and nodular small/medium-sized pleomorphic CD4(+) T cell infiltration. Cultures yielded Absidia corymbifera which was identified by phenotypic and molecular methods. Since a thorough examination did not detect organ involvement, the patient was diagnosed as having primary cutaneous zygomycosis. This is the first case report of cutaneous zygomycosis caused by A. corymbifera in a patient with primary cutaneous CD4(+) small/medium-sized pleomorphic T-cell lymphoma. Other cases of primary cutaneous zygomycosis caused by A. corymbifera are also reviewed. PMID- 19301176 TI - Algaemia in a dairy cow by Prototheca blaschkeae. AB - We describe the first known case of an algaemia by Prototheca blaschkeae in a dairy cow, which occurred after a chronic episode of mastitis caused by this pathogen. The organism was isolated from milk, joint fluid and blood samples, and microbiologic and molecular methods were performed to obtain a definitive identification of the algae. The affected cow was culled only after confirmation of a systemic infection by Prototheca. PMID- 19301177 TI - Detection of 2-pentylfuran in the breath of patients with Aspergillus fumigatus. AB - Aspergillus fumigatus produces 2-pentylfuran (2-PF) when cultured on blood agar, nutrient agar and other media. As 2-PF is not known to be produced by mammalian metabolism we hypothesized that it is detectable in breath of patients colonized or infected with A. fumigatus. Breath was tested for 2-PF from normal subjects, those undergoing chemotherapy, and adults at risk of colonization or infection with A. fumigatus because of bronchiectasis, cystic fibrosis, or immune suppression. Breath samples were collected in five L tedlar bags and analyzed by Gas Chromatography/Mass Spectroscopy (GC/MS) in MS-MS mode. 2-PF was not detected in breath 14 healthy controls, in one of 10 neutropenic subjects and 16 of 32 patients with lung disease. The sensitivity and specificity of the 2-PF breath tests when compared with recurrent isolation of aspergillus from sputum or from bronchoalveolar lavage over two months was 77% and 78% respectively. As 2-PF is not normally found in human breath its presence may reflect the active metabolism of A. fumigatus in the airways and form the basis of a new diagnostic breath test for Aspergillus infection. PMID- 19301178 TI - Metalloporphyrins, used for HO-1 inhibition, themselves affect hepatic microcirculation, liver function, and hepatocellular integrity. AB - BACKGROUND: Metalloporphyrins (MPs) are broadly used in the studies of the role of the heme oxygenase (HO)-1 system in different stress models. However, possible side effects of the MP administration itself have to be further investigated. METHODS: Sin IV mesoporpyhrin IX (SnMP; 10 micromol/kg body weight), tin protoporpyhrin IX (SnPP; 50 micromol/kg body weight), or chromium mesoporpyhrin IX (CrMP; 40 micromol/kg body weight) were administered to Sprague-Dawley rats (each group, n=5). The hepatic microcirculation was assessed by intravital microscopy (IVM). Blood samples were taken and the activity of HO-1 inhibition was measured by the determination of bilirubin accumulation after bile duct ligation. RESULTS: CrMP administration led to a decrease in mean arterial pressure. CrMP induced a marked hemolysis, a significant decrease of sinusoidal diameter and blood flow, and a marked inflammatory response. SnMP decreased sinusoidal diameters; however, this was compensated by an increase of sinusoidal red blood cell velocity. SnPP, but not SnMP, led to an increase of the number of nonperfused sinusoids. SnPP and CrMP revealed a two-fold increase in aspartite aminotransferase values after the completion of the IVM. The administration of MPs led to a 40%-50% decrease in the levels of conjugated bilirubin therapy, indicating that they actually inhibit HO-1 activity. CONCLUSIONS: The administration of MPs affects both the systemic macrohemodynamics and the hepatic microcirculation. As SnMP displayed the smallest number of side effects, this MP can be recommended for the studies of the HO-1 action on the liver microcirculation. PMID- 19301179 TI - Scaling laws for branching vessels of human cerebral cortex. AB - OBJECTIVE: Vascular architecture, particularly of cerebral microvessels, has profound implications for both health and disease in a variety of areas, such as neuroimaging, angiogenesis and development, Alzheimer's disease, and vascular tumors. We analyzed the architecture of tree-like vessels of the human cerebral cortex. METHODS: Digital three-dimensional images of the microvascular network were obtained from thick sections of India ink-injected human brain by confocal laser microscopy covering a large zone of secondary cortex. A novel segmentation method was used to extract the skeleton and measure the diameter at every vertex. RESULTS: In this paper, we focus on the topology of the cortical tree-like vessels. Using stem-crown decomposition, power-scaling laws were shown to govern the relationships between integrated parameters, such as the distal cumulative length, volume, or normalized flow. This led us toward an experimental confirmation of the allometric equation between mass and metabolic rate. Inversely, the power-law model did not match the relationships between local parameters, such as diameter, and integrated ones. As a consequence, Murray's law did not appropriately model the architecture of cerebrovascular bifurcations. CONCLUSIONS: This study provides a unique, large database and mathematical characterization that may prove valuable for modeling the cerebral. PMID- 19301181 TI - Abstracts from the XI Swedish Heart Association, April 22-24, 2009, Uppsala, Sweden. PMID- 19301183 TI - Proceedings of the XI International Orthoptic Congress in Antwerp, Belgium, May 2008. PMID- 19301184 TI - Preface for the Proceedings of the XI International Orthoptic Congress in Anwerp, May 2008. PMID- 19301185 TI - The influence of the vergence system on strabismus diagnosis and management. AB - The vergence system is an important element in human eye movement control. It comprises horizontal, vertical, and torsional components, the largest of which is convergence. Vergence performance is largely involuntary, although voluntary effort can influence convergence. Vergence function can be damaged by trauma or organic disease and, because convergence can be exerted voluntarily, it is susceptible to purposeful subjective disturbances. Vergence function is important in the maintenance of binocular control, being synonymous with motor fusion. The most common vergence dysfunction affects convergence and may be exhibited as an excess or weakness such as convergence spasm or paralysis. Divergence dysfunction generally presents with diminished function such as divergence weakness; vertical and cyclovergence disturbances are rare. An update is given on the neuroanatomical areas that subserve the vergence system. Clinical examples of the most common vergence disorders are presented. The influence of the vergence system on the diagnosis and management in common and less well-known clinical entities is described and discussed. PMID- 19301186 TI - A novel experimental method for measuring vergence and accommodation responses to the main near visual cues in typical and atypical groups. AB - Binocular disparity, blur, and proximal cues drive convergence and accommodation. Disparity is considered to be the main vergence cue and blur the main accommodation cue. We have developed a remote haploscopic photorefractor to measure simultaneous vergence and accommodation objectively in a wide range of participants of all ages while fixating targets at between 0.3 and 2 m. By separating the three main near cues, we can explore their relative weighting in three-, two-, one-, and zero-cue conditions. Disparity can be manipulated by remote occlusion; blur cues manipulated by using either a Gabor patch or a detailed picture target; looming cues by either scaling or not scaling target size with distance. In normal orthophoric, emmetropic, symptom-free, naive visually mature participants, disparity was by far the most significant cue to both vergence and accommodation. Accommodation responses dropped dramatically if disparity was not available. Blur only had a clinically significant effect when disparity was absent. Proximity had very little effect. There was considerable interparticipant variation. We predict that relative weighting of near cue use is likely to vary between clinical groups and present some individual cases as examples. We are using this naturalistic tool to research strabismus, vergence and accommodation development, and emmetropization. PMID- 19301187 TI - Emmetropization in anisometropic amblyopia. AB - PURPOSE: To investigate the process of emmetropization in anisometropic amblyopic subjects during childhood and to determine the effects of spectacle use only and patching. METHODS: Retrospectively, the records of patients who were diagnosed as anisometropic amblyopes were reviewed and 132 subjects who were between the ages of 5 to 8 at the time of diagnosis, with minimum follow-up of 3 years, were included in the study. Full ophthalmological examination was performed in every subjects and the information from every visit was retrieved for evaluation. Patients were divided into two groups according to the treatment they applied: patching and spectacle alone. RESULTS: The emmetropization process was similar to normal subjects after 3 years of follow-up, with a change of -0.71 D from the baseline spheric retinoscopy measurement in the spectacle group and -0.53 D in patching group (p > 0.809). Patching and spectacle use did not change the decrease in hypermetropia. Astigmatism and anisometropia scores did not show any difference in two groups in the 3-year follow-up period. CONCLUSION: The emmetropization process is similar to normal population in anisometropic subjects and spectacle use and patching therapy have no effects on this process. The anisometropia score does not change with patching and stays relatively the same during the follow-up in childhood. PMID- 19301188 TI - Factors affecting development of motor skills in extremely low birth weight children. AB - PURPOSE: The aim of this study is to analyze the impact of ophthalmic and neonatal factors on motor development in extremely low birth weight (ELBW) children. METHODS: Sixty-four ELBW children at least 3 years of age were recruited. Visual acuity (VA) was assessed using the Teller acuity cards (TACs) and a letter test, if possible. A validated questionnaire assessing 25 fine (part A) and 20 gross motor (part B) skills was administered to the parents. Data were collected on retinopathy of prematurity (ROP) zone, intraventricular haemorrhage (IVH), length of stay in hospital, and number of days on oxygen. RESULTS: Abnormal TAC acuity was associated with significantly lower scores on both parts A and B (part A: 21.5 versus 11.8, p < 0.001; part B: 17.5 versus 13.2, p < 0.001). Linear regression demonstrates a significant direct relationship between letter acuity and score A only (p = 0.03, r(2) = 0.179). Neither length of hospital stay, number of days ventilated, nor a history of IVH were associated with score A or B. However, the presence of ROP zone 1 was associated with a lower score A (p = 0.03). CONCLUSION: In this ELBW cohort VA and ophthalmic factors were the only factors associated with scores of development, particularly fine motor development. PMID- 19301189 TI - Visual perceptual consequences of stroke. AB - PURPOSE: Perceptual consequences of stroke include agnosia, alexia, dyschromatopsia, inattention, and hallucinations. Our aim was to evaluate these perceptual consequences in terms of prevalence after stroke. METHODS: Prospective multicenter cohort trial involving 14 recruiting centers (Vision in Stroke [VIS] group). Standardized referral and investigation protocol used by local investigators. Data presented from the first year of data collection, May 2006 to April 2007. Information obtained on visual acuity, ocular alignment and motility, visual field, visual inattention, and visual cognition/perception. RESULTS: One hundred seventy-eight patients excluded (49% male, 51% female) mainly due to inability to consent because of cognitive difficulties. Three hundred twenty three patients recruited (59% male, 41% female). Mean age at onset of stroke of 69 years (range 1-92). Median duration from onset to vision assessment of 22 days (range 0-2543 days). Type of stroke was an infarct in 79.5% and hemorrhagic in 20.5%. Six percent had had a previous stroke. Laterality of stroke was right sided in 48%, left sided in 40%, and bilateral in the remainder. Eight percent of patients had normal visual status; 68.4% had eye movement impairment, 46.1% had visual field impairment, and 25.1% had low vision; and 20.5% had perceptual difficulties. Of these, 14.2% had inattention, 1.3% had difficulty judging depth and distance, 0.3% had color detection problems, 2.5% complained of hallucinations, and 2.2% of agnosia. CONCLUSIONS: A substantial proportion (92%) of patients referred with suspected visual difficulty had visual impairment. One fifth of patients referred had perceptual consequences to their stroke relating to inattention and cortical visual processing impairment. PMID- 19301190 TI - A comparative analysis of monocular excursion measures. AB - PURPOSE: The range of monocular excursions can be determined by several different methods, including the Aimark Perimeter, Synoptophore and Modified Goldman Perimeter. However the relationship between these tests is poorly understood. METHODS: This is a prospective, cross-sectional study to compare the extent of monocular excursions by three methods. For each eye, the Goldman and the Synoptophore measured 8 axes, and the Aimark measured 12. Each assessment was timed. The results for direct elevation of the right eye are analyzed and comparisons made by the Bland-Altman analysis. RESULTS: One hundred and fourteen patients were recruited: 64 with abnormal eye movements and 50 with no ocular motility restrictions. The mean +/- SD rotations in degrees were Aimark 50.6 +/- 12.4, Synoptophore 24.6 +/- 5.78, and Goldman 40.6 +/- 9.14. The Bland-Altman plots showed the mean bias was highest when comparing the Synoptophore to the Aimark (-26 +/- 11.71) where the bias was inversely proportional to the rotation. The mean +/- SD times per axes in seconds were Aimark 16.38 +/- 4.62, Synoptophore 20.98 +/- 57.23, and Goldman 16.16 +/- 9.76. There was no significant difference in time taken between the tests (ANOVA, p = 0.50). CONCLUSION: There is considerable variation between these three methods for measurement of monocular excursions, but the difference in the time taken is accounted for by the variation in the number of axes assessed. PMID- 19301191 TI - Spontaneous consecutive exotropia in childhood. AB - PURPOSE: Consecutive exotropia is commonly the result of a surgical correction of a primary esotropia but may develop spontaneously. This study examines the clinical characteristics of patients who have developed a spontaneous consecutive exotropia and relates the key features to previously reported findings. METHODS: Patients were identified by routine clinical practice from May 2000 to December 2003 by retrospective case note analysis. Patients that presented with an esotropia that became an exotropia (without surgery or botulinum toxin A) were included. RESULTS: This study consisted of 14 patients who presented with an esotropia from early weeks of life to almost 4 years. No patients were undergoing occlusion treatment at the onset of the exotropia and no subject had dense amblyopia. The most common characteristics included (1) age of onset of esotropia of 24 months or less; (2) hypermetropic refractive error of +5 DS mean spherical equivalent; (3) lack of binocular single vision. CONCLUSION: The results highlight the variability of the condition with no single consistent finding, with the lack of binocular single vision, early onset esotropia, and high hypermetropia being common findings. Amblyopia was not found to be an important characteristic. PMID- 19301192 TI - Recurrence of intermittent exotropia: factors associated with surgical outcomes. AB - INTRODUCTION: The surgery of choice for intermittent exotropia (XT) continues to be debated, with little evidence that one procedure provides a more successful outcome than the other in the longer term. Many factors, however, may potentially influence the outcome of strabismus surgery for intermittent XT. This study aimed to investigate factors associated with the recurrence of an exo-deviation following horizontal muscle surgery for intermittent XT of the divergence excess type. METHODS: We retrospectively reviewed the medical histories of patients who underwent surgery for the correction of intermittent XT between January 1998 and June 2005. The factors analyzed were as follows: sex, age of onset, age at initial surgery, family history, size of preoperative deviation, near binocular single vision, amblyopia, oblique dysfunction, refractive error, type of surgery, and postoperative alignment. RESULTS: Eighty-nine patients were included in the final analysis. Of these, 19 demonstrated recurrence of their deviation. The mean follow-up period was 2 years. None of the factors analysed appeared to influence the outcome of intermittent XT surgery. CONCLUSION: We found that no single factor influenced patients' responses to the surgical treatment of intermittent XT. To address controversies and improve the evidence base regarding surgical intervention of this condition, randomized controlled trials are needed and justified because the results indicate that it would be relatively safe to randomly allocate patients to groups who could receive differing treatments so as to determine optimum management strategies. PMID- 19301193 TI - Midoperative forced duction test as a guide to titrate the surgery for consecutive exotropia. AB - This is a retrospective study of the outcome of all consecutive exotropia surgery performed between the years 1999 and 2007. Excluded were cases of slipped muscles, muscle transpositions, and concurrent cyclovertical procedures. A total of 135 patients of mean age 40 years (range 6-72) were operated with a mean preoperative angle of deviation -40Delta (range -12 to -90). Five were treated with fixed sutures and the rest with adjustables. The surgery included preplanned lateral rectus recessions of mean 6 mm (range 2-9) and maximal possible medial rectus advancements with or without resections of mean 5.6 mm (range 2-11), intraoperatively adjusted to give a resistance of +1 to +2 for abduction on forced duction test (FDT). Adjustable suture patients were seen 4 to 6 hours after surgery and underwent further adjustment, if needed, to achieve the target angle. A further review took place at 2 weeks and 2 months. Children remained under follow-up till 8 years of age. Forty percent needed postoperative adjustment. The mean postoperative angle at final measurement was -2Delta (range +25 to -45). Eighty percent achieved an angle within 10Delta of orthotropia. No gross restrictions of ductions were present postoperatively. PMID- 19301194 TI - Anterior ciliary circulation from the horizontal rectus muscles. AB - PURPOSE: To quantify the anterior ciliary arteries that accompany horizontal rectus muscles and to correlate these findings with analysis of arterial cross sectional area in distal tendons of these muscles. PATIENTS AND METHODS: In 31 consecutive patients, previously unoperated horizontal rectus muscles were photographed intraoperatively. Major and minor anterior ciliary arteries (ACAs) were counted from the photographs by a masked independent observer and compared between medial and lateral rectus muscles. Cross sections from medial and lateral rectus muscle resection specimens were stained for elastin to quantify arterial volume in the distal tendons. RESULTS: Twenty lateral and 22 medial rectus muscles from 31 patients were included. There was no significant difference in the mean number of major anterior ciliary arteries between lateral rectus muscles (2.25 +/- 0.79, range 1-4) and medial rectus muscles (2.05 +/- 0.65, range 1-3) (p = 0.46). The mean number of minor anterior ciliary arteries was 6.7 in both medial and lateral rectus muscles (p = 0.97). Resected rectus muscle tendons showed no significant difference between lateral and medial rectus muscles for arterial vessel number or cross-sectional area. CONCLUSIONS: These results show the contribution from the lateral rectus muscle to the anterior segment circulation may be more robust than is commonly taught. PMID- 19301195 TI - Convergence spasm treated with botulinum toxin. AB - PURPOSE: To analyze the effectiveness of botulinum toxin injection for convergence spasm. METHODS: Review of the Toxin Clinic database to identify patients who had undergone botulinum toxin injection to the medial rectus muscle for convergence spasm. RESULTS: There were 11 females and 6 males. Average age at treatment was 34 years (range 12-57 years). All patients had a variable esotropia, ranging from 10 to 90+ prism diopters but most could demonstrate some binocular function. All patients had convergence spasm, pupil miosis, and diplopia. Previous treatment included surgery, atropine, plus lenses, and occlusion. Fourteen patients (82%) were temporarily overcorrected after their first injection. Three patients (18%) were symptom free after just one injection and were subsequently discharged, all having been temporarily overcorrected. Seven patients (41%) continue with botulinum toxin on a regular basis, with or without occlusive contact lens, to remain symptom free, two being discharged after the third and fifth injections symptom free. Seven patients found no benefit from this treatment and so were discharged from the clinic or did not attend for follow-up appointments. CONCLUSION: Our rather extensive experience with these patients has led us to conclude that in some patients convergence spasm may be a type of dystonia, and may like other dystonias be suitable for botulinum toxin treatment. PMID- 19301196 TI - Patterns of word-list generation in mild cognitive impairment and Alzheimer's disease. AB - Patients with amnestic mild cognitive impairment (aMCI) have been described as exhibiting greater impairment on tests of category fluency than letter fluency. This has been offered as evidence that this condition represents pre-clinical Alzheimer's disease (AD). We hypothesized that this pattern of differential impairment is dependent on the specific semantic categories and initial letters selected, and is not specific to AD and aMCI. A total of 40 cognitively normal older adults, 74 MCI patients-25 "amnestic single domain" (aMCI), 27 "amnestic multiple domain", and 22 non-amnestic-and 29 AD patients were tested with multiple forms of semantic-category and initial-letter fluency tasks. The pattern of deficits within and across groups was highly dependent on the specific categories and letters chosen. Overall, aMCI patients did not demonstrate greater impairment in category than letter fluency. In fact, the level and pattern of their performance resembled that of cognitively normal older adults much more than AD patients. MCI patients with deficits in multiple cognitive domains performed most like AD patients. These findings indicate that verbal fluency performance is highly influenced by the specific tasks used, and impairment on semantic fluency is not characteristic of pure amnestic MCI. PMID- 19301197 TI - Role of human CYP1A1 and NAT2 in 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine induced mutagenicity and DNA adducts. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is carcinogenic in multiple organs and numerous species. Bioactivation of PhIP is initiated by PhIP N(2)-hydroxylation catalysed by cytochrome P450s. Following N-hydroxylation, O acetylation catalysed by N-acetyltransferase 2 (NAT2) is considered a further possible activation pathway. Genetic polymorphisms in NAT2 may modify cancer risk following exposure. Nucleotide excision repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human cytochrome P4501A1 (CYP1A1) and a single copy of either NAT2*4 (rapid acetylator) or NAT2*5B (slow acetylator) alleles were used to test the effect of CYP1A1 and NAT2 polymorphism on PhIP genotoxicity. Cells transfected with NAT2*4 had significantly higher levels of N hydroxy-PhIP O-acetyltransferase (p = 0.0150) activity than cells transfected with NAT2*5B. Following PhIP treatment, CHO cell lines transfected with CYP1A1, CYP1A1/NAT2*4 and CYP1A1/NAT2*5B each showed concentration-dependent cytotoxicity and hypoxanthine phosphoribosyl transferase (hprt) mutagenesis not observed in untransfected CHO cells. dG-C8-PhIP was the primary DNA adduct formed and levels were dose dependent in transfected CHO cells in the order: CYP1A1 < CYP1A1 and NAT2*5B < CYP1A1 and NAT2*4, although levels did not differ significantly (p > 0.05) following one-way analysis of variance. These results strongly support activation of PhIP by CYP1A1 with little effect of human NAT2 genetic polymorphism on mutagenesis and DNA damage. PMID- 19301198 TI - An antibody to IP-10 is a potent antagonist of cell migration in vitro and in vivo and does not affect disease in several animal models of inflammation. AB - IP-10 secretion is induced by pro-inflammatory cytokines and mediates the migration of CXCR3+ cells. Its elevation in clinical samples has been associated with multiple inflammatory diseases and its antagonism has been reported to be effective in several animal models of inflammatory disease. We generated a mouse anti-mouse IP-10 monoclonal antibody (mAb; Clone 20A9) that specifically bound murine IP-10 with high affinity and inhibited in vitro IP-10 induced BaF3/mCXCR3 cell migration with an IC(50) of approximately 4 nM. The 20A9 mAb was completely absorbed in vivo and had dose proportional pharmacokinetic exposure with a serum half life of 2.4-6 days. The 20A9 mAb inhibited IP-10 mediated T-cell recruitment to the airways, indicating that it is effective in vivo. However, administration of the 20A9 mAb had no significant effect on disease in mouse models of delayed type hypersensitivity, collagen induced arthritis, cardiac allograft transplantation tolerance, EAE or CD4+ CD45RBHi T-cell transfer-induced IBD. These data suggest that the 20A9 mAb can antagonize IP-10 mediated chemotaxis in vitro and in vivo and that this is insufficient to cause a therapeutic benefit in multiple mouse models of inflammatory disease. PMID- 19301200 TI - Follow-up of avidity and titer of anti-myeloperoxidase antibodies in sera from patients with primary ANCA-associated vasculitis. AB - Antineutrophil cytoplasmic antibodies (ANCA) are important serologic markers for ANCA-associated vasculitis (AAV). Our previous studies in propylthiouracil (PTU) induced AAV demonstrated that withdrawal of PTU resulted in clinical remission and significant decrease of avidity of PTU-induced myeloperoxidase (MPO)-ANCA. This study investigated the changes in avidity and titer of MPO-ANCA in sequential sera from some patients with primary AAV with different disease activities. Sequential sera samples of seven patients with MPO-ANCA-positive vasculitis at their initial onset, remission and relapse were collected. The avidity of MPO-ANCA was assessed by antigen-inhibition enzyme-linked immunosorbent assay (ELISA). The titer of MPO-ANCA was determined by a two-fold dilution of sera in MPO specific ELISA. The titer of MPO-ANCA was not significantly different between initial onset and remission. The avidity constant (aK) of MPO-ANCA in active phase is not significantly different from that in remission (724.9 +/- 828.4 l/mol vs. 353.4 +/- 551.7 l/mol, p = 0.303). No significant correlation could be found between aK and the level of Birmingham Vasculitis Activity Score, times of relapse, the number of organ involvement, serum creatinine, or CRP. Avidity and titer of MPO-ANCA did not decreased significantly during remission in AAV, indicating the chronic repeated antigen stimulation was not removed, which might be the reason for recurrent relapses. PMID- 19301199 TI - The receptor tyrosine kinase MerTK regulates dendritic cell production of BAFF. AB - The MerTK receptor tyrosine kinase is an important negative regulator of dendritic cell function and is required to prevent B cell autoimmunity in vivo. It is not currently known however, if any causal relationship exists between these two aspects of MerTK function. We sought to determine if dendritic cells (DC) from mice lacking MerTK (mertk(- / - ) mice) have characteristics that may aid in the development of B cell autoimmunity. Specifically, we found that mertk( / - ) mice contain an elevated number of splenic DC, and this population contains an elevated proportion of cells secreting the critical B cell pro survival factor, B cell activating factor (BAFF). Elevated numbers of BAFF secreting cells were also detected among mertk(- / - ) bone marrow-derived dendritic cell (BMDC) populations. This was observed in both resting BMDC, and BMDC stimulated with lipopolysaccharide (LPS) or treated with exogenous apoptotic cells. We also found that DC in general have a pro-survival effect on resting B cells in co-culture. However, despite containing more BAFF-secreting cells, mertk(- / - ) BMDC were not superior to C57BL/6 or baff-deficient BMDC at promoting B cell survival. Furthermore, using decoy receptors, we show that DC may promote B cell survival and autoimmunity through a BAFF-and a proliferation inducing ligand-independent mechanism. PMID- 19301201 TI - Effects of simvastatin on circulating autoantibodies to oxidized LDL antigens: relation with immune stimulation markers. AB - Statins exert a number of anti-inflammatory and immunomodulatory effects in vitro. However, the immunomodulatory effects in vivo are less clarified. In the present study, we investigated whether simvastatin treatment changed the levels of autoantibodies against specific oxidized LDL (oxLDL) antigens as well as their association with leukocyte activation markers. Eighty volunteers with mild-to moderate hypercholesterolemia were randomized to either simvastatin 40 mg or placebo for 6 weeks. Autoantibodies against apo B peptide antigens, C-reactive protein (CRP) and interleukin (IL)-6 in plasma were determined by ELISA. Subsets of circulating B and T cells were studied by flow cytometry. Simvastatin significantly reduced CRP by 26%, whereas IL-6 remained unchanged. Levels of IgG against the apo B peptide P-240 (amino acids 3586-3605) increased by 16% (p = 0.03) in the simvastatin group whereas autoantibody levels to other apo B peptides did not change. At baseline and after 6 weeks, the P-240 IgG levels were significantly correlated with the number of CD57+CD28 - CD8+T cells but not to other lymphocyte subsets or inflammatory markers. The P-240 IgG levels after 6 weeks simvastatin therapy was strongly correlated to the relative increase in CD57+CD28 - CD8+T cells (p = 0.003). Simvastatin treatment induced an increase in autoantibodies against an oxLDL antigen. The effect was related to an expansion of a CD8+T cell subset and may involve an immunostimulation by simvastatin. PMID- 19301202 TI - Carbonic anhydrase III is insufficient in muscles of myasthenia gravis patients. AB - Myasthenia gravis (MG) is considered as an autoimmune disease mainly mediated by antibodies against acetylcholine receptor. In recent years, other targets related to MG have been the subject of interest. Our previous research found that protein P25 was lower in muscles of MG patients using two-dimensional electrophoresis. In present study, anti-serum to P25 was prepared, immunohistochemistry and ATPase staining revealed that P25 was a muscle specific cytosolic protein and was mainly distributed in type I muscle fibers. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and precise molecular weight derived from mass spectrometer identified P25 as carbonic anhydrase III (CA III). Some members of CA family are related to autoimmune diseases and CA III is recently reported to be involved in rheumatoid arthritis. The results of immunoblot in this report showed that the level of CA III is specifically insufficient in the skeletal muscle of MG patients. The possible roles that CA III play in MG need further elucidation. PMID- 19301203 TI - Increased prevalence of polyomavirus BK viruria that correlates with thrombocytopenia in patients with systemic lupus erythematosus on intensive immunosuppressive therapy. AB - The prevalence of polyomavirus BK (BKV) reactivation is high in patients with systemic lupus erythematosus (SLE) on long-term immunosuppressants compared to normal population. However, only a few studies are available for the possible correlation of BKV reactivation and clinical manifestations in SLE patients. In the present study, we tried to correlate BKV viruria, clinical manifestations, laboratory findings, and medications in patients with SLE. The urine BKV viral DNA copies were detected from 95 patients with SLE and 32 healthy volunteers by real-time PCR. We found that the prevalence rate of BKV viruria in SLE patients was significantly higher than normal group (71.6% vs. 18.6%, p < 0.001) as well as the urine BKV DNA viral load (4.74 +/- 3.17 vs. 1.08 +/- 2.33 by log scale, p < 0.001). Interestingly, BKV viruria (+) SLE patients had more thrombocytopenic events than BKV viruria ( - ) group (32.4% vs. 3.7%, p = 0.008 after adjustment by age and sex). The patients with BKV viruria DNA copy number >3,200,000/ml exhibited more thrombocytopenia risk than BKV viruria <==3,200,000 copy number/ml or BKV viruria ( - ). The use of potent immunosuppressants may increase BKV viruria. In a refractory thrombocytoponeic case, the add-on of anti-BKV medication, leflunomide 20 mg/day rapidly decreased BKV viruria and recovered platelet counts. In conclusion, our study demonstrated that patients with SLE had higher prevalence rate of BKV reactivation that is correlated with thrombocytopenic episode. Intensive immunosuppressive therapy in SLE may increase the risk of BKV viruria. PMID- 19301204 TI - Multiple nuclear dots and rim-like/membranous IgG isotypes in primary biliary cirrhosis. AB - Anti nuclear (ANA) immunomorphological patterns such as multiple nuclear dots (MND) and rim-like/membranous (RL/M) are considered highly specific but little sensitive for primary biliary cirrhosis (PBC) diagnosis. To evaluate frequency and clinical significance of MND and RL/M in PBC patients when investigated at the level of immunoglobulin G isotypes. MND and RL/M pattern have been tested in 141 PBC sera and 230 pathological controls using HEp-2 cells as substrate and anti- total IgG and individual IgG subclasses (IgG1, IgG2, IgG3, IgG4) as specific antisera. One hundred and fourteen of 141 (80%) PBC patients had RL/M or MND pattern when IgG subclasses were used as revealing reagents (vs. 34% when anti total IgG were used, p < 0.0001). The prevalent isotype was IgG1 for RL/M, and IgG2 for MND pattern. None of controls was positive. No clinical differences in terms of severity and outcome of disease have been observed in PBC patients positive for MND and RL/M when investigated with IgG isotypes. The research for RL/M and MND pattern at level of IgG isotype determines a wide gain in terms of sensitivity without a loss of specificity. In Italian PBC patients MND and RL/M pattern did not seem to characterize any subgroup of patients with a poorer prognosis. PMID- 19301205 TI - Effects of interleukin-6 blockade on the development of autoimmune thyroiditis in nonobese diabetic mice. AB - We explored the role of interleukin-6 (IL-6) in the development of autoimmune thyroiditis in nonobese diabetic (NOD) mice, an animal model of Hashimoto's thyroiditis, using anti-mouse IL-6 receptor antibody (MR16-1). Thyroiditis was induced by iodide ingestion or mouse thyroglobulin (Tg) immunization. Mice were injected intraperitoneally with saline, control rat IgG, or MR16-1 (2 or 8 mg). Iodide ingestion did not increase serum IL-6 levels and MR16-1 (2 mg) failed to prevent the development of thyroiditis. In contrast, Tg immunization induced a rapid and significant increase in serum IL-6 levels. While MR16-1 (2 mg) had no effect on Tg-induced thyroiditis, the severity, but not incidence, of thyroiditis was reduced in 8 mg MR16-1-treated mice compared with saline-injected mice. However, thyroiditis development in the 8 mg MR16-1-treated mice was indistinguishable from that in the control IgG-treated mice. MR16-1 (8 mg) did not affect serum anti-Tg antibody levels. These results suggest that IL-6 may play only a minor role in the development of autoimmune thyroiditis in NOD mice. PMID- 19301206 TI - The susceptibility to experimental autoimmune encephalomyelitis is not related to dysferlin-deficiency. AB - Recent observations suggested that dysferlin might play a role in the development of autoimmune central nervous system (CNS) inflammation. To address this issue, we studied the induction and effector phase of experimental autoimmune encephalomyelitis in C57BL/10 mice producing intact or functionally deficient dysferlin. We found that both types of mice showed identical T-cell and antibody responses against the immunogen, and developed CNS inflammation with identical clinical courses, frequencies, lesion distributions, sizes and compositions. These findings suggest that the presence or absence of dysferlin does not have any consequences for the triggering or effector phase of autoimmune CNS inflammation. PMID- 19301207 TI - AEB-071 has minimal impact on onset of autoimmune diabetes in NOD mice. AB - Protein kinase C (PKC) is an important signaling enzyme in the activation and regulation of T lymphocytes. T-cell-mediated destruction of beta-cells is a characteristic feature of autoimmune (Type 1) diabetes. Here we explore the ability of PKC inhibition, using the PKC inhibitor AEB-071 (AEB), to reduce disease in two animal models of spontaneous autoimmune diabetes (non-obese diabetic (NOD) mouse and biobreeding rat (BB)). NOD mice were treated with AEB for 4 weeks, starting at either 4 weeks of age (prior to the development of insulitis) or at 8 weeks of age, once insulitis is present. Animals treated with AEB during the effector phase of the disease (treatment onset at 8 weeks of age), showed a 2-week delay in diabetes onset (p < 0.05). In these animals, the extent of insulitis was lower than in vehicle-treated controls; however, neither serum autoimmune anti-GAD65 antibody levels nor pancreatic insulin content were different between experimental groups. Overall, inhibition of PKC can mildly reduce lymphocytic infiltrate of pancreatic islets and modestly delay onset of autoimmune diabetes in NOD mice. AEB, a T-cell-targeted immunosuppressive strategy, is only sufficient as a monothereapy to modestly delay onset of autoimmune disease in the NOD mouse. PMID- 19301208 TI - The antifibrogenic effect of hepatocyte growth factor (HGF) on renal tubular (HK 2) cells is dependent on cell growth. AB - Although several reports suggest an antifibrogenic effect of hepatocyte growth factor (HGF), an increased deposition of matrix induced by HGF has also been reported. These conflicting effects could result from a diverse proliferative state of the target cells. Aim of the present study was to evaluate HGF effects on growth arrested (quiescent) and actively proliferating renal tubular epithelial (HK-2) cells. HK-2 cells were cultured in RPMI medium either on agarose gel or on plastic surface in order to inhibit or to allow cell proliferation. Cells were incubated with RPMI containing HGF (50 ng/ml) for 24 h at 37 degrees C. Untreated HK-2 were used as control. After 24 h of incubation, cells were counted by Coulter counter. (alpha2)IV collagen, transforming growth factor-beta (TGF-beta), Tissue inhibitor of metalloproteases (TIMP1 and 2) mRNA levels were determined by RT-PCR. The production of type IV collagen, c-met, proliferating cell nuclear antigen (PCNA), and SnoN, a transcriptional Smad corepressor and thus a TGF-beta inhibitor, was evaluated by ELISA or western blotting. MMP-9 and 2 gelatinolytic activity was studied by zymography. Treatment with HGF did not increase HK-2 cell number and PCNA synthesis when the cells were grown on agarose as it did for cells grown on plastic surface. HGF increased (alpha2)IV collagen in proliferating cells whereas it reduced (alpha2)IV collagen and c-met synthesis in growth arrested cells. HGF treatment increased TGF-beta and TIMP-2 in proliferating cells while reduced TIMP-1 mRNA levels of quiescent cells. Furthermore, production of the co repressor SnoN was significantly decreased by HGF in proliferating cells. Quiescent and proliferating HK-2 showed a different pattern of metalloproteases activity with a prevalence of MMP2 in quiescent and MMP9 in proliferating cells. In summary, HGF showed opposite effects on growth arrested and proliferating HK-2 cells favouring matrix deposition in the latter with increasing expression of collagen, TIMP-1 and TGF beta. Our results demonstrate that the proliferative state of target cells may influence the effects of HGF on extracellular matrix turnover in HK-2 cells. PMID- 19301209 TI - Orthographic facilitation in oral vocabulary acquisition. AB - An experiment investigated whether exposure to orthography facilitates oral vocabulary learning. A total of 58 typically developing children aged 8-9 years were taught 12 nonwords. Children were trained to associate novel phonological forms with pictures of novel objects. Pictures were used as referents to represent novel word meanings. For half of the nonwords children were additionally exposed to orthography, although they were not alerted to its presence, nor were they instructed to use it. After this training phase a nonword picture matching posttest was used to assess learning of nonword meaning, and a spelling posttest was used to assess learning of nonword orthography. Children showed robust learning for novel spelling patterns after incidental exposure to orthography. Further, we observed stronger learning for nonword-referent pairings trained with orthography. The degree of orthographic facilitation observed in posttests was related to children's reading levels, with more advanced readers showing more benefit from the presence of orthography. PMID- 19301211 TI - Athletic performance and weight changes during the "Marathon of Sands" in athletes well-trained in endurance. AB - The purpose of this study is to examine the effects of the "Marathon of Sands" (MS), a 7-day, self-sufficient-diet, multi-stage running race across a section of the Moroccan desert, on body weight and plasma volume variation (PVV) and the relationship of these factors to performance in athletes who are well-trained in endurance. Sixteen MS runners agreed to participate in this study. Weight and body composition were measured and venous blood samples were taken before the first stage (D0), after the third stage (D3) and at the end of the MS (after the sixth stage: D6). Haematocrit and haemoglobin were used to calculate PVV at (D0, D3, and D6). No significant plasma volume decrease was observed throughout the race. Significant decreases in total body weight (BW), fat-free mass (FFM) and fat mass (FM) were observed in D3 and D6 (-4.3%, -3.5%, -0.8%; and -6.1%, -5%, 1.1%, respectively, for BW, FFM and FM at D3 and D6). This study clearly shows that, despite extreme conditions, the MS did not lead to a significant PV decrease in athletes well-trained in endurance. This study also supports the hypothesis that significant body weight loss may not systematically affect performances during long duration multiple-stage races. PMID- 19301212 TI - Respiratory muscle training in athletes with spinal cord injury. AB - The effect of respiratory muscle endurance training (RMET) on RM function, dyspnoea and exercise performance was evaluated in SCI athletes. Nine endurance athletes (7 paraplegics T4-L1, 2 post-polio syndromes) were evaluated on three occasions (T1-T3), with a 1-month interval between evaluations. Participants performed between T1 and T2 their standard individual exercise training program (control), and between T2 and T3 the same program with 5 additional RMET sessions per week. Each evaluation included: lung function tests, RM strength and endurance tests, a maximal incremental arm cranking test and a field test (simulated competition). Ventilation and dyspnoea were evaluated during each exercise test. Lung function variables and maximal inspiratory strength were not modified (p>0.05) while maximal expiratory strength (+23+/-36 cm H2O; p<0.01) and respiratory endurance (+3 min 33 s+/-2 min 42 s, p<0.01) increased from T2 to T3. During the arm cranking test, exercise duration and maximal power output were slightly increased at T3 compared to T2 (+46+/-39 s, p=0.09 and +8+/-8 W, p=0.08) while ventilation and dyspnoea remained similar. During the field test, exercise time (-10+/-33 s, p=0.37) and ventilation were unchanged but dyspnoea was reduced (-2+/-2 pts, p=0.02) between T2 and T3. We concluded that RMET can improve RM function, reduce the perception of dyspnoea but modifies only slightly exercise performance in SCI athletes. PMID- 19301213 TI - How well do skinfold equations predict percent body fat in elite soccer players? AB - The use of generic equations for estimating percent body fat from skinfold thicknesses can be criticised when applied to specific sports. The present aims were to compare existing methods of using skinfold data and to derive an equation for predicting body fat values in professional soccer players. Forty-five professional soccer players (24.2 +/- 5.0 years; 82.0 +/- 8.5 kg; 1.82 +/- 0.07 m) participated. Skinfold thicknesses were assessed at eight sites for the application of existing prediction equations. Skinfold data were also utilised to determine a novel soccer-specific equation. All players had a reference estimate of percent fat by dual-energy x-ray absorptiometry (DXA). The existing skinfold equations differed from the DXA-referenced values by varying degrees, the equation of Withers et al. (1987) demonstrating the lowest bias and highest relationship and agreement with DXA. Regression analysis resulted in an equation incorporating anterior thigh, abdominal, triceps and medial calf sites, accounting for 78.4% variance in DXA criterion values. PMID- 19301214 TI - DEXA body composition changes among 140 conscripts. AB - The aim of the study was to determine changes in body composition and physical fitness during military service. A prospective cohort study of 140 healthy male conscripts was conducted. We examined subject characteristics, aerobic performance and muscle strength, and assessed body composition using dual-energy X-ray absorptiometry (DEXA) three times. Conscripts' mean baseline weight (79.5 kg) decreased by 2 kg during the first 3 months, but increased by 0.9 kg during the second 3-month period (p<0.001). Fat mass measured by DEXA decreased by 3.2 kg during the first but increased by 0.8 kg during the second 3-month period (p<0.001). Throughout the 6-month study, an increase was seen in distance of 12 min run test (from 2 380 m to 2 530 m; p<0.001), and muscle strength score (from 6.5 to 9.5 p<0.001). Finnish military training seems to have beneficial effects on physical fitness. However, considering the relatively modest changes in body fat and physical fitness seen in conscripts with average BMIs at baseline, design of diverse training programmes for the varying baseline BMI levels are warranted to improve the physical fitness results. PMID- 19301215 TI - Physical demands of top-class soccer assistant refereeing during high-standard matches. AB - This study examined the physical demands experienced by top-class assistant referees during high-standard football matches. Computerized match analyses and heart rate recordings were performed on 18 international assistant referees during the FIFA Confederations Cup 2005. Total distance covered averaged 5752+/ 554 m, of which 34% was covered at a high-intensity (>13 km.h (-1)). Sidewards movements represented 30% of total match distance. Both total distance (r=0.67; P<0.01) and high-intensity activities (r=0.52; P<0.05) were related to the displacements of the ball in the same match. The distance covered by high-speed running in this tournament was 255% greater (P<0.001) than that covered during the under-17 World Championship 2003. Mean heart rate during the game was 140+/ 11 b.min (-1) (78+/-4% HR (max)). Mean heart rate during a 5-min interval of the match was related in part (r=0.31; P<0.001) to the amount of high-intensity activities performed within the same match-period. The results of this study show that the physical demands imposed on assistant referees are influenced by the movements of the ball and the standard of the competition. PMID- 19301216 TI - Skin-fold thickness and training volume in ultra-triathletes. AB - We investigated the relationship between variables of anthropometry and training volume on race performance in 29 male non-professional ultra-triathletes. Anthropometric variables were determined in order to calculate body mass index, sum of skin-folds and percent body fat. Participants kept a comprehensive training diary recording their training volume in hours and kilometres in the 3 months before the race. The relationship of anthropometry and average weekly training volume with race performance was investigated with linear regression analysis. The sum of 8 skin-fold thicknesses was associated with total race time (r (2)=0.33, p<0.001), whereas the average weekly training volume was not (r (2)=0.00, p>0.05). The training volume showed no association with the sum of 8 skin-folds (r (2)=0.00, p>0.05). The sum of 8 skin-folds was neither associated with speed in the swim (r (2)=0.10, p>0.05) nor in the bike split (r (2)=0.10, p>0.05) but showed a significant association with speed in the run split (r (2)=0.38, p<0.0001). We concluded that anthropometry was of more importance than training volume in male Triple Iron triathletes and that these athletes were close to runners regarding the relationship of anthropometry with race performance. PMID- 19301217 TI - Psychophysiological stress response during competition between elite and non elite Korean junior golfers. AB - The purpose of this study was to investigate the effects of stress hormone and competition state anxiety response between elite and non-elite junior golfers in competition. Six elite (Handicap: 2.67+/-0.82; 16.2+/-1.38 yrs) and six non-elite (Handicap: 7.83+/-1.17; 15.8+/-0.75 yrs) Korean junior golfers participated in this study. Physiological stress and psychological stress responses were measured on four occasions (at rest, prior to, during, after competition) by salivary cortisol and Competitive State Anxiety Inventory-2 (CSAI-2) respectively. In salivary cortisol, no interaction was found between groups and the test occasions. However, both groups showed significantly increased levels between rest and all other occasions and between prior to and after competition. The interpretation of this finding is limited by the interaction with the diurnal rhythm of cortisol secretion. In cognitive state anxiety, significant interaction was found. Elite golfers had significantly lower cognitive state anxiety scores than non-elite golfers at rest, prior to and during competition. No interaction was shown in somatic state anxiety. However, there was significant difference among occasions. Both groups had higher somatic state anxiety scores prior to and during competition than after competition. No interaction was shown in self confidence, but elite group had significantly higher scores than non-elite group at all occasions. This study demonstrates the differences of psychophysiological response in competition between elite golfers and non-elite golfers. PMID- 19301218 TI - Isokinetic thigh muscle ratios in youth football: effect of age and dominance. AB - This study aimed to analyse the effect of age on muscle peak torque (PT), and Hamstring (H): Quadricep (Q) ratio in elite youth footballers. To date, no study has considered age-group playing level and pubertal development in this population. One hundred and fifty-seven elite youth footballers in the age groups U12 to U18 volunteered to participate in this study, 133 of these were further grouped for pubertal development. Prior to testing subjects completed separate familiarisation, a three minute cycle ergometer warm up (resistance 50-60W), and two sub-maximal repetitions. Concentric and eccentric isokinetic PT measures for reps 2-4 of H and Q muscle action were taken at 60 degrees s (-1). From this, conventional and functional H: Q ratio was calculated along with dominant: non dominant ratio for the concentric Q and H, and eccentric H conditions. Significant main effects were observed for the age/pubertal development group and PT in all muscles and conditions (P<0.05). Of particular interest was a significant main effect for age and Functional H: Q (P<0.05), which suggested a move away from equality at U18. Our study provides normative data for coaches, trainers and clinicians working with youth footballers and may also have connotations for injury prevention and performance. PMID- 19301219 TI - Fitness in contemporary dance: a systematic review. AB - It has been suggested that dancers are less fit compared to other athletes. However, the majority of studies make their arguments based on data deriving mainly from ballet. Therefore, the aim of the current review was to investigate: a) aerobic and anaerobic fitness, muscular strength and body composition characteristics in contemporary dancers of different levels, and b) whether supplementary exercise interventions, in addition to normal dance training, further improves contemporary dance performance. Three databases (Medline, Cochrane and the Cumulative Index to Nursing & Allied Health research database) were searched to identify publications regarding the main fitness components of contemporary professional and student dancers. At a professional level, it appears that contemporary dancers demonstrate higher maximal oxygen uptake and higher scores in muscular endurance than ballet dancers. However, contemporary dance students are equally fit compared to their ballet counterparts and their body composition is also very similar. Only two studies have investigated the effects of supplementary exercise training on aspects of dance performance. Further research is needed in order to confirm preliminary data, which suggest that the implementation of additional fitness training is beneficial for contemporary dance students to achieve a better performance outcome. PMID- 19301220 TI - Endurance training of moderate intensity increases testosterone concentration in young, healthy men. AB - The aim of this study was to investigate the effect of short-term, moderate intensity and low volume endurance training on gonadal hormone profile in untrained men. Fifteen young, healthy men performed an endurance training of 5 week duration on a cycle ergometer. Before and after the exercise program all participants completed a maximal incremental test. Concentration of testosterone (T), sex hormone-binding globulin (SHBG) and cortisol (C) as well as blood morphology were determined in venous blood samples at rest both before and after the training. The training program resulted in 3.7% improvement of maximal oxygen uptake (VO(2max)) and 8.2% improvement of power output reached at VO(2max) (PO (max)). This was accompanied by significant increase in T (from 18.84+/-5.73 nmol.l(-1) to 22.03+/-6.61 nmol.l(-1), p = 0.0004) and calculated fT concentration (from 374+/-116 pmol.l(-1) to 470+/-153 pmol.l(-1), p = 0.00005). Moreover, the training caused a significant decrease in SHBG concentration (from 34.45+/-11.26 nmol.l(-1) to 31.95+/-10.40 nmol.l(-1), p = 0.01), whereas no significant changes were found in the cortisol concentration (334+/-138 nmol.l( 1) vs. 367+/-135 nmol.l(-1) for pre- and post-training measures, respectively, p = 0.50) and T/C and fT/C ratios. We have concluded that short-term, moderate intensity and low volume endurance training can significantly increase testosterone concentration in previously untrained men. PMID- 19301221 TI - Effect of intensive interval training during unloading on muscle deoxygenation kinetics. AB - The present study investigated the effects of intensive interval training during 20-day of unloading on local muscle oxygenation kinetics evaluated by near infrared spectroscopy technique (NIRS). Eleven adult men completed 20-day unloading and were divided into two groups; the control (CON) group and training (TR) group. The TR group engaged in exercise training sessions that consisted of one-legged submaximal cycle exercise using the unloaded leg at 60 approximately 80% of VO(2peak) with intermittent rest periods, 25 min/day every other day. All subjects performed isometric knee extension exercise at 50% of their maximum voluntary contraction force before and after unloading. NIRS Delta[deoxy-Hb/Mb] signal was recorded from m. vastus lateralis and was fitted to an exponential equation in order to determine the kinetics parameters. The time constant (tau) of the % Delta[deoxy-Hb/Mb] was unchanged in the TR group, while it significantly increased in the CON group after unloading (pre, 5.0+/-1.0; post, 7.4+/-1.0 s). It is concluded that 20-day unloading increased the tau, suggesting deterioration of capacity for oxidative phosphorylation and oxygen utilization in a skeletal muscle. Additionally, the preservation of tau in the TR group suggested that intensive interval training could have an impact on the maintenance of muscle oxidative metabolism during unloading. PMID- 19301222 TI - Association of GNB3 C825T polymorphism with peak oxygen consumption. AB - The C825T single nucleotide polymorphism (SNP) in the guanine nucleotide-binding protein, beta polypeptide 3 ( GNB3) gene gives rise to a splice variant, GNB3s that has enhanced G protein activation and signal transduction activity. This variant has been reported to be associated with cardiovascular disease, diabetes and obesity. We studied this SNP in 95 healthy 18 to 30 year-old African American university students to determine its association with aerobic capacity and cardiorespiratory fitness as measured by peak oxygen consumption (VO (2)peak). We also tested the effect of heart rate variability (HRV) as an independent predictor of VO (2)peak. We tested the association of the SNP and HRV with VO (2)peak in a multivariate regression analysis with appropriate adjustments of covariates, under dominant and recessive models. We found a significant independent association of the 825T allele with VO (2)peak under the dominant model (beta-coef.=-0.101, P=0.0442). We also observed that HRV marginally influenced VO (2)peak. This finding suggests that GNB3 C825T polymorphism is associated with VO (2)peak which is influenced by autonomic modulation of heart rate in African Americans. PMID- 19301223 TI - Heart rate variability and ventilatory efficiency. AB - The aims of this investigation were to identify possible associations between the resting high frequency component of heart rate variability and measures of ventilatory efficiency, determined during exercise, in a group of endurance trained athletes. V'E, V'CO2, and V'O2 were measured during incremental exercise up to the anaerobic threshold (AT) and up to volitional exhaustion (V'O2 max) in 12 subjects. Resting high frequency heart rate variability (HF(NU)), determined using spectral analysis of ECG traces, was correlated with the V'E vs. V'CO2 slope calculated from data up to the AT, where a negative linear relation between the V'E vs. V'CO2 slope and HF (NU) was recorded (R(2)=0.69). Also, resting HF (NU) was correlated with the V'E vs. V'CO (2) slope, when the data used for slope calculation included all V'E vs. V'CO2 data up to V'O2 max (R(2)=0.56). However, there was no evidence of association between the minimum V'E:V'CO (2) and resting HF(NU) (R(2)=0.08). Negative relations between V'E vs. V'CO2 slope and resting HF(NU) suggest that HF(NU) plays a role in matching blood flow in the lung to alveolar ventilation throughout the ventilatory cycle. PMID- 19301224 TI - Relationship between the MRI and EMG measurements. AB - The purpose of this study was to investigate the effect of intensive eccentric exercise on hamstring muscles by using magnetic resonance imaging (MRI) and to elucidate the relationships between the changes in the electromyographic (EMG) parameters and in the transverse relaxation time (T2) of the hamstring muscles. Seven male volunteers performed eccentric knee flexion exercise, and the EMG activity of the hamstring muscles was simultaneously measured. Before and immediately after the exercise, the maximum isometric knee flexion torque was measured and MR images of the hamstring muscles were obtained. For all hamstring muscles, the EMG activity of the fifth set was significantly lower than that of the first set. For each subject, a significant correlation was detected between the percentage change in the value of the post-exercise T2 value and those of EMG signals during the exercise only for the semitendinosus (ST) muscle and not for the biceps femoris (BF) and the semimembranosus (SM) muscles. These results suggested that the EMG-activity reductions in the BF, ST, and SM muscles were due to neuromuscular fatigue, and moreover the reduction in the ST muscle was due to a failure in the E-C coupling, which was caused by excessive muscle-fiber damage. PMID- 19301225 TI - MMP-2 is not altered by stretching in skeletal muscle. AB - Considering that short bouts of stretching, as recommended in rehabilitation and sports activities, induce skeletal muscle and connective tissue adaptation, the hypothesis of this study was that MMP-2 activity is regulated by muscle stretch. The level of MMP-2 activity was, thereby, assessed after stretching in rat soleus muscle. Animals received a single session of stretching (10 stretches lasting 1 min each with 30 s of rest in between) and were evaluated immediately and after 8, 24, 48, 72 and 168 h. To evaluate the effect of repetitive sessions of stretching, three groups of animals were evaluated - one group after 2 sessions, another after 3, and a third after 7. MMP-2 activity was evaluated by zymography and MMP-2 mRNA was assessed by real-time polymerase chain reaction. None of the groups presented MMP-9 activity. MMP-2 activity and mRNA expression did not change after either single or repetitive sessions of stretching. In conclusion, the results of this study indicate that MMP-2 is not involved in the muscle stretch-induced remodeling. PMID- 19301226 TI - Medical management of extremely low-birth-weight infants in the first week of life: a survey of practices in the United States. AB - We sought to determine the current practices of neonatologists in their management of extremely low-birth-weight (< 1000 g) infants. We directly mailed an anonymous survey to the medical directors of 809 neonatal intensive care units in the United States. More than one-third of those surveyed responded, with a substantial majority from intensive care (level III) nurseries or extracorporeal membrane oxygenation centers. Academic centers and private practice environments were both well represented. Some traditional practices have changed, such as beginning resuscitation with 40% rather than 100% oxygen. Many practices vary based on whether neonates are cared for in private versus academic centers, including initial resuscitation method, type of ventilation used, use of intraventricular hemorrhage prophylaxis, and routine antibiotic therapy. Parenteral nutrition composition and the use of inhaled nitric oxide differ based on the responding center's participation in clinical trials. The number of years in practice as a neonatologist does not affect practice decisions. Among all our findings, the prevalence of one potentially harmful practice, the continued use of dexamethasone for corticosteroid therapy, was particularly noteworthy. In conclusion, the strength of evidence does not always predict whether practices are adopted or abandoned. Further research is necessary to clarify the optimal management for this high-risk patient population. PMID- 19301227 TI - Attitudes and practices regarding use of progesterone to prevent preterm births. AB - We sought to describe current attitudes and practices of obstetrician gynecologists regarding use of progesterone and prevention of preterm birth. A self-administered survey was mailed to American College of Obstetricians and Gynecologists Fellows and Junior Fellows in Practice in March to May 2007. The survey consisted of 36 questions, including respondents' demographic characteristics, preterm birth risk factor knowledge and screening practices, and use of progesterone for the prevention of preterm birth. The response rate was 52% ( N = 345); most respondents were general obstetrician-gynecologists (89%). Many (74%) reported recommending or offering progesterone for prevention of preterm birth. Almost all (93%) reported use for the indication of previous spontaneous preterm birth. However, many also reported use for other indications such as dilated/effaced cervix (37%), short cervix on ultrasound (34%), and cerclage (26%). These results suggest that most obstetricians recommend or offer progesterone to prevent preterm birth for women with a previous spontaneous preterm birth and many also offer it for women with other high-risk obstetric conditions. PMID- 19301228 TI - Fat and cholesterol diet induced lipid metabolic disorders and insulin resistance in rabbit. AB - BACKGROUND: Lipid disorder has been found to result in insulin resistance (IR). IR often is associated with other cardiovascular risk factors. However, the pathogenesis of human IR is not completely understood. METHODS: The present study was designed to examine if rabbits were fed with a diet containing high fat and high-cholesterol diet (HFCD) could develop lipid disorder and subsequently IR. Male Japanese white rabbits were fed either a normal chow diet or HFCD for 20 weeks. Plasma levels of triglycerides (TG), total cholesterol (TC), glucose, and insulin were measured. To evaluate glucose metabolism, we performed an intravenous glucose tolerance test. In addition, we compared adipose tissue accumulation and aortic atherosclerosis lesions in HFCD-fed rabbits with those in control rabbits. RESULTS: In HFCD-fed rabbits there was an increase in plasma levels of TC and TG as well as visceral adipose tissue accumulation. Severe aortic atherosclerotic lesions were found in HFCD-fed rabbits. Although there were no differences in body weight, plasma insulin and blood pressure between the two groups, HFCD-fed rabbits showed higher insulin IR index compared to control rabbits. CONCLUSION: Our results showed that HFCD induced IR, increased adipose accumulation and atherosclerotic lesions in rabbits, suggesting that the HFCD-fed rabbits can serve as a model for the research on human IR and lipid metabolism abnormalities. PMID- 19301229 TI - Mutations in the melanocortin 4 receptor (MC4R) gene in obese patients in Norway. AB - BACKGROUND: Mutations in the melanocortin 4 receptor ( MC4R) gene are the most frequent cause of monogenic forms of obesity, but the reported prevalences of mutations in obese individuals diverge, varying from 0.2 to 5.8%. OBJECTIVE: The aim of this study was to assess the prevalence of MC4R mutations in obese children and adults residing in Norway. SUBJECTS AND METHODS: We sequenced the coding region of MC4R in 1 027 obese patients. Among these, were 644 adults with a BMI >35 kg/m(2) and 383 children with a body weight >97.5 percentile for height. Identified mutations were analyzed by family studies and a bioinformatic approach including comparative sequence analysis and prediction of impact on transmembrane helix and three dimensional structure. RESULTS: Nine mutations were identified, of which four were novel and five previously described. The prevalence of MC4R mutations was 1.6% in pediatric and 0.8% in adult patients. All four novel mutations, I69R, M79I, I195S, and M200del were identified among pediatric patients. M79I was found in an ethnic Norwegian patient, while the rest were identified in second generation immigrants. The previously described mutations Y35X/D37V, V95I, T150I, R236C and V253I were identified in one pediatric and five adult patients. None of the adult patients reported childhood onset of obesity. The M200del mutation was found in a homozygous state, while the rest were heterozygous. CONCLUSION: MC4R mutations are not a common cause of obesity in Norway and screening of obese patients does not appear to be warranted. The results are consistent with results from previous studies. PMID- 19301230 TI - ACE-inhibitor suppresses the apoptosis induced by endoplasmic reticulum stress in renal tubular in experimental diabetic rats. AB - AIMS: The aim of the present study was to investigate the role of the endoplasmic reticulum stress and apoptosis in experimental diabetic nephropathy. The effects of ACE inhibitor on the endoplasmic reticulum stress and apoptosis were also assessed. METHODS: Diabetes was induced in male Sprague-Dawley rats by injection with streptozotocin at 60 mg/kg i .p. Diabetic rats were then randomly assigned into control (untreated) or treatment of an ACE inhibitor, perindopril, for 24 weeks. Tubulointerstitial injury was assessed by histopathology. Tubule apoptosis was detected by TUNEL assay. Endoplasmic reticulum stress associated proteins expression of glucose-regulated protein-78 (GRP78/BiP), phospho-eukaryotic initiation factor 2 alpha (eIF2 alpha), phospho-pancreatic ER kinase (PERK) and caspase-12 was assessed by immunohistochemistry and Western blots. RESULTS: There were more TUNEL-positive nuclei in diabetic kidneys than in control kidneys. At 24 weeks, experimental diabetes was associated with a considerable increase in protein expression of GRP78, phospho-eIF2 alpha, phospho-PERK, and caspase-12 in the tubulointestitium. ACE inhibitor not only attenuated the apoptosis but also reduced the overexpression of these endoplasmic reticulum stress associated proteins in tubulointestitium of diabetic rats. CONCLUSIONS: Increased tubular apoptosis in experimental diabetic rats is attenuated by blockade of the renin angiotensin system with an ACE inhibitor, which might be in an association with reduced endoplasmic reticulum stress. PMID- 19301231 TI - Continuous subcutaneous insulin infusion versus multiple daily insulin injections in type 2 diabetes: a meta-analysis. AB - BACKGROUND: Continuous Subcutaneous Insulin Infusion (CSII) improves HbA1c in type 1 diabetic patients unsatisfactorily controlled by Multiple Daily Injections (MDI). Few trials have explored CSII for basal-bolus therapy in type 2 diabetes. MATERIALS AND METHODS: Randomized Clinical Trials (RCTs) comparing CSII and MDI for at least 12 weeks in type 2 diabetic patients were retrieved, assessing between-group differences in HbA1c and insulin daily dose at endpoint, and incidence of hypoglycemia. RESULTS: A total of 4 RCTs was included in the analysis. CSII did not produce any significant improvement of HbA1c in comparison with MDI (Standardized difference in mean: 0.09(-0.08;0.26)%; p=0.31). No significant difference was observed in the rate of hypoglycemic episodes. CSII was associated with a nonsignificant trend toward the reduction of insulin doses used at the end of trial. CONCLUSIONS: Available data do not justify the use of CSII for basal-bolus insulin therapy in type 2 diabetes. PMID- 19301232 TI - Effect of two starting insulin regimens in patients with type II diabetes not controlled on a combination of oral antihyperglycemic medications. AB - In an open-label, 24-week, parallel-group study, 135 patients inadequately controlled with oral antihyperglycemic medications (OAMs) were treated with maximally tolerated doses of metformin and glibenclamide for at least 8 weeks and then randomized to bedtime neutral protamine Hagedorn (NPH) insulin plus maximally tolerated dose of glibenclamide BID (glib/NPH group) or insulin lispro mix 50 (50% lispro, 50% insulin lispro protamine suspension [ILPS]) pre-breakfast and lispro mix 25 (25% lispro, 75% ILPS) pre-dinner (LM50/LM25 group) (both OAMs discontinued). The LM50/LM25 group had significantly lower 2-hour postprandial BG (both meals combined) compared with glib/NPH after 12 (11.70+/-3.40 mmol/L vs. 13.15+/-2.44 mmol/L, p=0.010) and 24 weeks (11.13+/-3.31 mmol/L vs. 14.46+/-2.93 mmol/L, p =0.0001). Both regimens significantly decreased HbA1c. The reduction was greater with LM50/LM25 (-1.31+/-2% vs. -0.5+/-1.6%; P=0.01). At endpoint, the overall hypoglycemia rate increased with LM50/LM25 and decreased with glib/NPH compared with baseline (0.22+/-0.9 vs. -0.08+/-0.72 episodes/patient/30 days; p =0.037). Treatment with LM50/LM25 compared with glib/NPH in patients with inadequate control on combined OAMs yielded better postprandial and overall glycemic control with a higher rate of hypoglycemia. PMID- 19301233 TI - Changes in the use of analogue insulins in 37 206 children and adolescents with type 1 diabetes in 275 German and Austrian centers during the last twelve years. AB - AIMS: To describe changes in insulin treatment in children and adolescents in Germany/Austria regarding short acting (SA)- and long acting (LA) insulin analogues in different age groups over the last twelve years. METHODS: Use of different insulins and treatment regimens were analysed in 37 206 children and adolescents with the age of 0-20 years from 276 German and Austrian centers that were registered in the DPV-database (Dec. 2007). The group was subdivided into 4 age groups (A: 0-4 years; B:5-9 years;C:10-14 years,D:15-19 years). We further analysed the use of analogues from onset of diabetes. RESULTS: A significantly increasing percentage of pediatric patients in all age groups with Type 1 Diabetes use analogue insulins. In 2007, 48.5% used SA, 45.8% LA. 87.2% of pumps were running with short acting analogue. Age specific analysis: A: 2000: 9.1%SA, 0.7%LA vs. 2007: 50.0%SA, 10.0%LA; B: 2000:5.6%SA, 1.4%LA vs. 2007: 36.8%SA, 27.5%LA C: 2000:14.3%SA, 3.4%LA vs. 2007: 45.3%SA, 49.1%LA D: 2000:26.3%SA, 3.2%LA vs. 2007: 59.1%SA, 61.9%LA. This increase in usage of analogues was also found at onset of diabetes. Corrected for age, center and diabetes duration HbA1c was significantly lower in the group with regular insulin (8.18+/-0.047%) than with SA (8.32+/-0.048%) (p<0.0001) and BMI-SDS was only marginal lower in the group with regular insulin (0.45+/-0.01 kg/m (2)) than with SA (0.47+/-0.15) (p>0.007). Similar differences in HbA1c (8.09+/-0.05% vs. 8.40+/-0.05%) and BMI SDS where seen when NPH was compared with LA respectively. After change to SA the reduction of severe hypoglycemia with (6.1/100 pat.years) and without coma (6.2/100 pat.years) was significant and after change to LA for severe hypoglycemia without coma respectively. CONCLUSIONS: Long-term data for the use of new drugs are sparse. In our analysis patients are followed not under study conditions. Still the higher BMI and HbA1c with either SA or LA usage have to be discussed carefully in the context of increasing use of both, long acting and short acting analogues and possible problems with reimbursement. PMID- 19301234 TI - Peripheral nerve injuries: an international survey of current treatments and future perspectives. AB - Peripheral nerve injuries are a serious health concern and leave many patients with lifelong disabilities. There is little information about incidences, current practice, outcomes, and type of research that may help delineate new strategies. A questionnaire was designed to determine characteristics of peripheral nerve injuries and the need for alternative strategies and sent to 889 plastic, hand, trauma, and orthopedic surgeons in 49 countries; 324 completed surveys were collected and analyzed (total response rate of 36.45%). The majority of institutions treat more than 3000 patients annually. Trauma was the leading cause of injury with the majority located on the upper extremity. In most cases, a primary repair was achieved, but 2.52% were unrepairable. The overall outcome was linked to their Sunderland classification (SCL). A grade 1 nerve injury (SCL-1) reached a maximum outcome after 7.15 months. SCL-2, -3, -4, and -5 needed 10.69, 14.08, 17.66, and 19.03 months, respectively. Tissue engineering was considered the most important research field, resulting in a visual analogue scale of 8.6. Despite marked advances in the treatment of peripheral nerve injuries, clinical outcomes still appear unsatisfactory. The importance of research in the field of tissue engineering should be emphasized as a pathway toward improving these outcomes. PMID- 19301235 TI - Venous flow-through flap reconstruction following severe finger wound infection: case report. AB - We report a case of a 22-year-old patient who sustained a box-cutter injury to the index finger on her right hand complicated by severe felon. Extensive tissue debridement left the patient with a defect exposing the underlying flexor tendon. A venous flow-through flap from the right forearm was used to provide coverage, which survived completely without venous congestion. Excellent functional and cosmetic result was achieved with this approach. Because of its similarity in color and non-hair-bearing nature, the venous flow-through flap serves as an excellent approach for volar digital defect coverage in which the underlying flexor tendon is exposed. PMID- 19301236 TI - Free tissue transfer in the private practice setting: a review of 100 patients. AB - Although free tissue transfer is a well-established method of reconstruction for difficult tissue defects, the technique is not yet widely embraced in the private practice setting. Reasons for this include concern over the possibility of flap loss and other complications, the time required for these procedures, and the perceived need for a microsurgical team. The results of free tissue transfer in 100 patients in a single practitioner private practice setting were assessed. Charts were reviewed and data recorded including major and minor complications, American Society of Anesthesia rating, operative time, intensive care unit and postoperative hospital stay, estimated blood loss, and transfusion requirements. Indications included breast reconstruction, the management of chronic wounds, and acute trauma. There were three complete flap losses and three partial flap losses. There were 22 major complications in 21 patients and 17 minor complications. The results of this study compare favorably with those compiled from 36 prior reports. Free tissue transfer has taken its place in the reconstructive armamentarium as the procedure of choice in many reconstructive situations due to its reliability and versatility. This form of reconstruction can be safely offered to patients in any practice situation by surgeons who are trained in and comfortable with free tissue transfer techniques. PMID- 19301237 TI - [Legal consequences in cases of deaths in police custody]. AB - From 1993 to 2003, 128 people died in police custody in Germany. The autopsy records allowed a more detailed analysis in 60 of these cases. The most frequent cause of death was cerebro-cranial trauma followed by alcohol, drug and medication poisoning. 24 (40%) of those 60 cases led to preliminary proceedings against responsible personnel such as doctors, police officers, paramedics and a hospital nurse. Prosecution later dropped proceedings in 75% of these cases in accordance with Section 170 Subsection 2 of the German Code of Criminal Procedure since there was no criminal conduct. In 2 of the cases, charges were dropped on condition. Only one of these 24 cases led to a conviction by an order of summary punishment. This case study discusses the legal assessment in the light of the high burden of proof required in a criminal case in Germany. PMID- 19301238 TI - Risk of clinically relevant bleeding in warfarin-treated patients--influence of SSRI treatment. AB - PURPOSE: To investigate the risk of clinically relevant bleeding in warfarin treated patients with or without concomitant treatment with selective serotonin reuptake inhibitors (SSRIs). METHODS: A cohort study was performed in patients treated with warfarin due to atrial fibrillation. Exposed patients were defined as patients treated with SSRI at any time between January 1999 and September 2005 (n = 117). Unexposed patients without SSRI-treatment were randomly selected and matched for age and sex (1:1). The primary endpoint was hospital admission due to bleeding during the same time period. RESULTS: Bleeding occurred in 17 exposed patients (totally 23 bleedings) and in two unexposed patients (totally two bleedings) (p = 0.0003). A total of 11 bleedings occurred during treatment with the combination of warfarin and SSRI, and 14 during treatment with warfarin only. The total incidences of bleedings per 1000 treatment years were 51.4 (25.7-92.0) and 23.9 (13.1-40.1), respectively, and the unadjusted incidence rate ratio (IRR) 2.15 (0.88-5.11). Cox regression analysis including first bleedings revealed an adjusted hazard ratio of 3.49 (1.37-8.91) for bleeding during treatment with a combination of SSRI and warfarin compared with treatment with warfarin only. Initiation of SSRI therapy was not associated with a change in dose of warfarin or with a change in international normalized ratio (INR) (p = 0.48 and p = 0.31, respectively). CONCLUSION: Addition of SSRI to warfarin-treated patients may be associated with an increased risk of clinically relevant bleeding. The effect seems not to be associated with a direct influence of SSRI on the anti-coagulant activity of warfarin. PMID- 19301239 TI - High-resolution power doppler sonography in inflammatory myopathy. PMID- 19301240 TI - Comparison of loudness adjustments by MCL and maplaw in users of the MED-EL COMBI 40/40+ cochlear implant system. AB - The programming of a CI speech processor involves measurement of electrical threshold and 'most comfortable loudness' (MCL) levels on each electrode. However, when the derived 'map' is activated the resultant percept is often too loud or too soft. In this situation, adjustments of MCL settings are usually made in order to achieve a comfortable percept.The present study examined the range of volume control adjustments in 24 users of the MED-EL C40/40+ device. Volume control settings varied from 36% to 126% with a mean of 84.3% (SD = 20.4%). Additionally, the output compression function ('maplaw') was adjusted to test whether this alternative manipulation could result in a comfortable percept. It was found that this could be achieved satisfactorily, with maplaw values ranging from 10 to 2000. Clinical implications of these findings are discussed. PMID- 19301241 TI - Enhancing well-being and alleviating depressive symptoms with positive psychology interventions: a practice-friendly meta-analysis. AB - Do positive psychology interventions-that is, treatment methods or intentional activities aimed at cultivating positive feelings, positive behaviors, or positive cognitions-enhance well-being and ameliorate depressive symptoms? A meta analysis of 51 such interventions with 4,266 individuals was conducted to address this question and to provide practical guidance to clinicians. The results revealed that positive psychology interventions do indeed significantly enhance well-being (mean r=.29) and decrease depressive symptoms (mean r=.31). In addition, several factors were found to impact the effectiveness of positive psychology interventions, including the depression status, self-selection, and age of participants, as well as the format and duration of the interventions. Accordingly, clinicians should be encouraged to incorporate positive psychology techniques into their clinical work, particularly for treating clients who are depressed, relatively older, or highly motivated to improve. Our findings also suggest that clinicians would do well to deliver positive psychology interventions as individual (versus group) therapy and for relatively longer periods of time. PMID- 19301243 TI - Effects of cellulase and xylanase enzymes on the deconstruction of solids from pretreatment of poplar by leading technologies. AB - Comparative data is presented on glucose and xylose release for enzymatic hydrolysis of solids produced by pretreatment of poplar wood by ammonia fiber expansion (AFEX), ammonia recycled percolation (ARP), controlled pH, dilute acid, flowthrough (FT), lime, and sulfur dioxide (SO(2)) technologies. Sugar solubilization was measured for times of up to 72 h using cellulase supplemented with beta-glucosidase at an activity ratio of 1:2, respectively, at combined protein mass loadings of 5.8-116 mg/g of glucan in poplar wood prior to pretreatment. In addition, the enzyme cocktail was augmented with up to 11.0 g of xylanase protein per gram of cellulase protein at combined cellulase and beta glucosidase mass loadings of 14.5 and 29.0 mg protein (about 7.5 and 15 FPU, respectively)/g of original potential glucose to evaluate cellulase-xylanase interactions. All pretreated poplar solids required high protein loadings to realize good sugar yields via enzymatic hydrolysis, and performance tended to be better for low pH pretreatments by dilute sulfuric acid and sulfur dioxide, possibly due to higher xylose removal. Glucose release increased nearly linearly with residual xylose removal by enzymes for all pretreatments, xylanase leverage on glucan removal decreased at high cellulase loadings. Washing the solids improved digestion for all pretreatments and was particularly beneficial for controlled pH pretreatment. Furthermore, incubation of pretreated solids with BSA, Tween 20, or PEG6000 prior to adding enzymes enhanced yields, but the effectiveness of these additives varied with the type of pretreatment. PMID- 19301244 TI - Cyclopamine-induced synophthalmia in sheep: defining a critical window and toxicokinetic evaluation. AB - Cyclopamine, a steroidal alkaloid, from the plant Veratrum californicum is teratogenic, causing a range of different birth defects. The critical window for cyclopamine-induced synophthalmia formation has been reported to be gestational day (GD) 14. The objectives of this study were to better describe cyclopamine induced craniofacial deformities, to better define the window of susceptibility to synophthalmia formation, and to characterize cyclopamine toxicokinetics in sheep. Ewes were dosed i.v. with purified cyclopamine for toxicokinetic analysis. Another four groups of ewes were dosed orally twice daily with 0.88 g/kg of V. californicum on GD 13, 14 or 15 or consecutively on GD days 13-15. Pregnancy and pre-partum fetal malformations were determined by ultrasound imaging on GD 60. At parturition lambs were assessed for gross malformations. The elimination half life of cyclopamine in ewes was determined to be 1.1 +/- 0.1 h. The rapid clearance of cyclopamine indicates that ingestion of V. californicum must occur during a very narrow window for synophthalmia formation to occur. Ewes dosed with V. californicum on GD 13 or 14 had lambs with various craniofacial malformations including cyclopia, maxillary dysplasia and mandibular micrognathia. Ewes dosed on GD 15 delivered normal lambs. Ewes dosed consecutively on GD 13-15 were not pregnant at GD 60 and Veratrum-induced embryonic death was assumed to be the cause. Interestingly, lambs with cyclopia were smaller, under-developed and appeared premature even though their twin appeared fully developed. Initial evaluations suggest this was due to placental dysplasia. PMID- 19301245 TI - Antiapoptotic effects of dietary antioxidants towards N-nitrosopiperidine and N nitrosodibutylamine-induced apoptosis in HL-60 and HepG2 cells. AB - The aim of this work was to determine the effect of vitamin C, diallyl disulfide (DADS) and dipropyl disulfide (DPDS) towards N-nitrosopiperidine (NPIP) and N nitrosodibutylamine (NDBA)-induced apoptosis in human leukemia (HL-60) and hepatoma (HepG2) cell lines using the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay. None of the vitamin C (5-50 microm), DADS and DPDS (1-5 microm) concentrations selected induced a significant percentage of apoptosis. In simultaneous treatments, vitamin C, DADS and DPDS reduced the apoptosis induced by NPIP and NDBA in HL-60 and HepG2 cells (around 70% of reduction). We also investigated its scavenging activities towards reactive oxygen species (ROS) produced by NPIP and NDBA using 2',7' dichlorodihydrofluorescein diacetate in both cell lines. ROS production induced by both N-nitrosamine was reduced to control levels by vitamin C (5-50 microm) in a dose-dependent manner. However, DADS (5 microm) increased ROS levels induced by NPIP and NDBA in HL-60 (40 and 20% increase, respectively) and HepG2 cells (18% increase), whereas DPDS was more efficient scavenger of ROS at the lowest concentration (1 microm) in both HL-60 (52 and 25% reduction, respectively) and HepG2 cells (24% reduction). The data demonstrated that the scavenging ability of vitamin C and DPDS could contribute to inhibition of the NPIP- and NDBA-induced apoptosis. However, more than one mechanism, such as inhibition of phase I and/or induction of phase II enzymes, could be implicated in the protective effect of dietary antioxidants towards NPIP- and NDBA-induced apoptosis in HL-60 and HepG2 cells. PMID- 19301246 TI - Massive ovarian oedema: an unusual abdominal mass in infancy. AB - We describe a 13-month-old female who presented with vaginal bleeding, breast and pubic hair development and an abdominal mass. She underwent emergency laparotomy and left-sided salpingoophorectomy. Histological examination of the resected ovary revealed massive ovarian oedema, a rare non-neoplastic enlargement of the ovary. Consideration of this diagnosis in patients with an abdominal mass and endocrine disturbance may allow conservative surgery and preservation of fertility. PMID- 19301247 TI - Treatment of an infant with severe acute refractory immune thrombocytopenic purpura using combination therapy including rituximab. AB - Some infants with acute immune thrombocytopenic purpura (ITP) do not respond to first-line therapy, and currently there is no consensus on therapy for these refractory cases. We describe a 12-week-old infant with acute ITP who was unresponsive to intravenous immunoglobulin and corticosteroid, and developed gastrointestinal bleeding. Several combination therapies were unsuccessful. After four doses of rituximab followed by intravenous immunoglobulin and corticosteroid, his platelet counts gradually increased. Combined therapy which includes rituximab may be a promising treatment for severe acute refractory ITP. PMID- 19301248 TI - Mouse x pig chimeric antibodies expressed in Baculovirus retain the same properties of their parent antibodies. AB - The development of hybridoma and recombinant DNA technologies has made it possible to use antibodies against cancer, autoimmune disorders, and infectious diseases in humans. These advances in therapy, as well as immunoprophylaxis, could also make it possible to use these technologies in agricultural species of economic importance such as pigs. Porcine reproductive and respiratory syndrome virus (PRRSV) is an arterivirus causing very important economic losses to the industry. Passive transfer of antibodies obtained by biotechnology could be used in the future to complement or replace vaccination against this and other pig pathogens. To this end, we constructed and studied the properties of chimeric mouse x pig anti-PRRSV antibodies. We cloned the constant regions of gamma-1 and gamma-2 heavy chains and the lambda light chain of pig antibodies in frame with the variable regions of heavy and light chains of mouse monoclonal antibody ISU25C1, which has neutralizing activity against PRRSV. The coding regions for chimeric IgG1 and IgG2 were expressed in a baculovirus expression system. Both chimeric antibodies recognized PRRSV in ELISA as well as in a Western-blot format and, more importantly, were able to neutralize PRRSV in the same fashion as the parent mouse monoclonal antibody ISU25C1. In addition, we show that both pig IgG1 and IgG2 antibodies could bind complement component C1q, with IgG2 being more efficient than IgG1 in binding C1q. Expressing chimeric pig antibodies with protective capabilities offers a new alternative strategy for infectious disease control in domestic pigs. PMID- 19301249 TI - A high efficient method of constructing recombinant Bombyx mori (silkworm) multiple nucleopolyhedrovirus based on zero-background Tn7-mediated transposition in Escherichia coli. AB - A high efficient way for generation of recombinant Bombyx mori (silkworm) multiple nucleopolyhedrovirus by Tn7-mediated transposition in Escherichia coli was performed. The new system consists of a conditional replication donor vector pRCDM and an attTn7 site blocked E. coli containing BmNPV-Bacmid. The donor vector contains a replication origin derived from R6Kgamma, which propagated only in host cells with pir gene expression decreased in the transposition background greatly. Compared with original vector derived from pUC, the transposition efficiency increased from 5.7 to 66% ( approximately 10 fold) when using conditional replication vector pRCDM transposition into original BmDH10Bac. A further effort to decrease the transposition background was made by blocking the attTn7 site in host E. coli genome. The resulting attTn7 occupied BmDH10BacDeltaTn7 resulted in a significant increase from 5.7 to 23% ( approximately 4 fold) in the efficacy of generate recombinant BmNPV Bacmid by transposition. Furthermore, the transposition of BmDH10BacDeltaTn7 with pRCDM resulted typically in 100% white colonies, and it indicated that a zero transposition background was accomplished. This high efficient and zero background transposition system provides a new simple and rapid method for construction of recombinant BmNPV used to express target genes or produce gene delivery virus particles in silkworm. PMID- 19301250 TI - Modality specific functional interaction in sensorimotor synchronization. AB - Movement execution strongly relies on precise sensorimotor synchronization. In a finger-tapping task that requires subjects to synchronize their finger taps to regular pacing signal synchronization accuracy varies with respect to pacing signal's modality. This study aimed at elucidating functional brain dynamics associated with modality specific behavioral synchronization accuracy. To this end, 10 right-handed subjects performed a finger-tapping task with respect to regular auditory and visual pacing, respectively, whereas neuromagnetic activity was recorded using a 122-channel whole-head neuromagnetometer. Visual pacing was associated with significantly reduced tap-to-pacer asynchrony and increased intertap variability as compared to auditory pacing. The brain dynamics associated with task execution were analyzed using the frequency domain beamformer approach dynamic imaging of coherent sources (DICS). Both tasks were shown to be associated with comparable networks. However, during visual pacing involvement of the ventral premotor cortex (PMv) was shown, whereas during auditory pacing the dorsal premotor cortex (PMd) was concerned with task execution. Synchronization with respect to visual pacing was associated with significantly increased functional interaction between thalamus and PMv at beta frequency as compared to functional interplay between thalamus and PMd during auditory pacing. Auditory synchronization was associated with increased functional interaction between left superior temporal gyrus and PMd at alpha frequency. Furthermore, functional interaction between thalamus and premotor cortex at beta frequency was significantly correlated with synchronization accuracy. All in all the present data suggest that modality specific synchronization differences are associated with frequency and connectivity specific changes of functional interaction in distinct brain networks. PMID- 19301251 TI - Self-reported health and physician diagnosed illnesses in women with posttraumatic stress disorder and major depressive disorder. AB - Posttraumatic stress disorder has been associated with poor physical health. Depression is also associated with poor health, and may be responsible for the apparent relationship between PTSD and health outcomes. The current study examined self-reported and physician diagnosed medical morbidity in women. Women with PTSD alone were compared to three other groups of women: women with PTSD and comorbid major depressive disorder (MDD), women with MDD only, or women with neither diagnosis (comparison group). Results suggest that PTSD, with or without MDD, is associated with poor health in women. Posttraumatic stress disorder severity was related to health complaints beyond the effects of both somatization and depressive symptoms among women with PTSD. Findings and implications are discussed in relation to previous research in the area. PMID- 19301252 TI - Anxiety sensitivity and aspects of alexithymia are independently and uniquely associated with posttraumatic distress. AB - Using a sample of adult survivors of physical trauma requiring hospitalization (N = 677), we examined the relationship of aspects of alexithymia and anxiety sensitivity to symptoms of posttraumatic distress (PTD). At the bivariate level, both aspects of alexithymia and anxiety sensitivity were positively associated with acute PTD symptomatology, but anxiety sensitivity was more strongly related to PTD symptoms. At the multivariate level, both anxiety sensitivity and aspects of alexithymia made unique and independent contributions to both total PTD symptoms and the majority of PTD symptom clusters. At the facet level, anxiety sensitivity-physical concerns and anxiety sensitivity-psychological concerns, and the alexithymic dimension of difficulty identifying feelings, were uniquely associated with acute PTD symptoms. Findings are discussed in terms of potential clinical implications. PMID- 19301253 TI - Pleural mesothelioma: an unusual case diagnosed on pleural fluid cytology and immunocytochemistry. AB - Mesothelioma is a rare neoplasm with relationship to occupational and environmental exposure to asbestos. Its accurate and early diagnosis is often difficult. We present an unusual clinical presentation and diagnostic dilemma in a 30-year-male, who presented with neck pain and diffuse edema of left upper limb. The color Doppler ultrasound revealed venous thrombosis. The right supraclavicular lymph node biopsy revealed a poorly differentiated carcinoma. The patient had mild bilateral pleural effusion, the characteristic cytomorphological features of mesothelioma on fluid cytology were helpful in establishing the diagnosis. PMID- 19301254 TI - Plant-produced human growth hormone shows biological activity in a rat model. AB - Plants have been shown to be efficient systems for expressing a wide range of recombinant proteins from various origins. Here, using a plant virus-based expression vector to produce human growth hormone (hGH) in Nicotiana benthamiana plants, we demonstrate, for the first time, that the plant-produced hGH (pphGH) is biologically active in a hypophysectomized rat model. We observed an average weight gain of approximately 17 g per animal in a group of 10 animals that were injected subcutaneously with pphGH with 60 microg/dose for 10 days. With the increasing demand for hGH, accompanied with the need to make this recombinant protein available to a wider population at a more reasonable cost, plants provide a feasible alternative to current production platforms. PMID- 19301255 TI - Secreted frizzled related protein 1 is a target to improve fracture healing. AB - Genetic studies have identified a high bone mass of phenotype in both human and mouse when canonical Wnt signaling is increased. Secreted frizzled related protein 1 (sFRP1) is one of several Wnt antagonists and among the loss-of function mouse models in which 32-week-old mice exhibit a high bone mass phenotype. Here we show that impact fracture healing is enhanced in this mouse model of increased Wnt signaling at a physiologic level in young (8 weeks) sFRP1( /-) mice which do not yet exhibit significant increases in BMD. In vivo deletion of sFRP1 function improves fracture repair by promoting early bone union without adverse effects on the quality of bone tissue reflected by increased mechanical strength. We observe a dramatic reduction of the cartilage callous, increased intramembranous bone formation with bone bridging by 14 days, and early bone remodeling during the 28-day fracture repair process in the sFRP1(-/-) mice. Our molecular analyses of gene markers indicate that the effect of sFRP1 loss-of function during fracture repair is to accelerate bone healing after formation of the initial hematoma by directing mesenchymal stem cells into the osteoblast lineage via the canonical pathway. Further evidence to support this conclusion is the observation of maximal sFRP1 levels in the cartilaginous callus of a WT mouse. Hence sFRP1(-/-) mouse progenitor cells are shifted directly into the osteoblast lineage. Thus, developing an antagonist to specifically inhibit sFRP1 represents a safe target for stimulating fracture repair and bone formation in metabolic bone disorders, osteoporosis and aging. PMID- 19301256 TI - FGF2 stimulates SDF-1 expression through the Erm transcription factor in Sertoli cells. AB - Ets-related molecule (Erm) is a member of the Ets transcription factor family. Erm is known to be an important factor for the self-renewal of Spermatogonial stem cells (SSCs) and the maintenance of spermatogenesis. We investigated the molecular mechanism of Erm regulation on SDF-1 in TM4 Sertoli cells. Erm and Sdf 1 levels were up-regulated after FGF2 treatment in TM4 cells, whereas these levels were significantly decreased by FGF2 in ST2 bone marrow stromal cells. Knockdown of Erm by siRNA in the presence of FGF2 decreased the Sdf-1 levels in TM4 cells. The expression levels of Erm were similar and Erm overexpression increased the Sdf-1 in both TM4 and ST2 cells. FGFR subtype analysis revealed that FGFR4 was expressed in TM4 cells but not in ST2 cells. A blocking experiment also confirmed that FGFR4 is partly responsible for the up-regulation of Erm and SDF-1 induced by FGF2 stimulation in TM4 cells. FGF2 and ERM increased the activity of Sdf-1 gene promoter region in a dose-dependent manner. EMSA revealed that ERM strongly binds to the -846 to -851 nucleotide region of the potential Ets binding site (EBS) in the Sdf-1 promoter. In addition, CXCR4, the SDF-1 receptor, was expressed in spermatogonia and Sertoli cells in the seminiferous tubules of the mouse testis. Our results indicate that ERM directly regulates Sdf 1 gene expression by interacting with its cis-acting element in response to FGF2 stimulation in TM4 cells. PMID- 19301257 TI - Osteoblastic bone formation is induced by using nanogel-crosslinking hydrogel as novel scaffold for bone growth factor. AB - Bone regeneration for the defects in revision surgery of joint replacement is an increasingly important issue. To repair bone defects, bone cell activation by growth factors using synthetic resorbable scaffold is a useful and safe option. We examine the efficiency of nanogel-crosslinking hydrogel as a novel synthetic scaffold for BMP to stimulate osteoblasts and to induce bone formation. Cholesterol-bearing pullulan nanogel-crosslinking hydrogel (CHPA/Hydrogel) was used to deliver BMP. The CHPA hydrogel pellets were implanted in vivo. Single implantation of CHPA/hydrogel containing low amounts of BMP induced osteoblastic activation and new bone formation in vivo. Furthermore, nanogel in a disc shape established recruitment of osteoblastic cells that vigorously formed bone to heal the calvarial defects, which did not heal spontaneously without it. In conclusion, CHPA/hydrogel serves as an efficient and versatile scaffold for the stimulation of osteoblasts to form bone and to repair defects via delivery of BMP. PMID- 19301258 TI - Asynchronous DNA replication and origin licensing in the mouse one-cell embryo. AB - To prevent duplicate DNA synthesis, metazoan replication origins are licensed during G1. Only licensed origins can initiate replication, and the cytoplasm interacts with the nucleus to inhibit new licensing during S phase. DNA replication in the mammalian one-cell embryo is unique because it occurs in two separate pronuclei within the same cytoplasm. Here, we first tested how long after activation the oocyte can continue to support licensing. Because sperm chromatin is licensed de novo after fertilization, the timing of sperm injection can be used to assay licensing initiation. To experimentally skip some of the steps of sperm decondensation, we injected mouse sperm halos into parthenogenetically activated oocytes. We found that de novo licensing was possible for up to 3 h after oocyte activation, and as early as 4 h before DNA replication began. We also found that the oocyte cytoplasm could support asynchronous initiation of DNA synthesis in the two pronuclei with a difference of at least 2 h. We next tested how tightly the oocyte cytoplasm regulates DNA synthesis by transferring paternal pronuclei from zygotes generated by intracytoplasmic sperm injection (ICSI) into parthenogenetically activated oocytes. The pronuclei from G1 phase zygotes transferred into S phase ooplasm were not induced to prematurely replicate and paternal pronuclei from S phase zygotes transferred into G phase ooplasm continued replication. These data suggest that the one-cell embryo can be an important model for understanding the regulation of DNA synthesis. PMID- 19301259 TI - CTGF enhances migration and MMP-13 up-regulation via alphavbeta3 integrin, FAK, ERK, and NF-kappaB-dependent pathway in human chondrosarcoma cells. AB - Tumor malignancy is associated with several features such as proliferation ability and frequency of metastasis. Connective tissue growth factor (CTGF), a secreted protein that binds to integrins, modulates the invasive behavior of certain human cancer cells. However, the effect of CTGF on migration activity in human chondrosarcoma cells is mostly unknown. Here we found that CTGF increased the migration and expression of matrix metalloproteinase (MMP)-13 in human chondrosarcoma cells (JJ012 cells). RGD peptide, alphavbeta3 monoclonal antibody (mAb) and MAPK kinase (MEK) inhibitors (PD98059 and U0126) but not RAD peptide inhibited the CTGF-induced increase of the migration and MMP-13 up-regulation of chondrosarcoma cells. CTGF stimulation increased the phosphorylation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK). In addition, treatment of JJ012 cells with NF-kappaB inhibitor (PDTC) or IkappaB protease inhibitor (TPCK) inhibited CTGF-induced cell migration and MMP-13 up regulation. Stimulation of JJ012 cells with CTGF also induced IkappaB kinase alpha/beta (IKK alpha/beta) phosphorylation, IkappaBalpha phosphorylation, p65 Ser(536) phosphorylation, and kappaB-luciferase activity. The CTGF-mediated increases in kappaB-luciferase activities were inhibited by RGD, PD98059, U0126 or FAK, and ERK2 mutant. Taken together, our results indicated that CTGF enhances the migration of chondrosarcoma cells by increasing MMP-13 expression through the alphavbeta3 integrin, FAK, ERK, and NF-kappaB signal transduction pathway. PMID- 19301260 TI - Model-free feedback design for a mixed cancer therapy. AB - In this article, a model-free feedback control design is proposed for the drug administration in mixed cancer therapy. This strategy is very attractive because of the important issue of parameter uncertainties unavoidable when dealing with biological models. The proposed feedback scheme use past measurements to update an on-line simplified model. The control design is then based on model predictive control in which a suitable switching is performed between two different cost functions. The effectiveness of the proposed model-free control strategy is validated using a recently developed model (unknown to the controller) governing the cancer growth on a cells population level under combined immune and chemotherapy and using real human data. PMID- 19301261 TI - Mechanistic study of saikosaponin-d (Ssd) on suppression of murine T lymphocyte activation. AB - Saikosaponin-d (Ssd) is a triterpene saponin derived from the medicinal plant, Bupleurum falcatum L. (Umbelliferae). Previous findings showed that Ssd exhibits a variety of pharmacological and immunomodulatory activities including anti inflammatory, anti-bacterial, anti-viral and anti-cancer effects. In the current study we have investigated the effects of Ssd on activated mouse T lymphocytes through the NF-kappaB, NF-AT and AP-1 signaling pathways, cytokine secretion, and IL-2 receptor expression. The results demonstrated that Ssd not only suppressed OKT3/CD28-costimulated human T cell proliferation, it also inhibited PMA, PMA/Ionomycin and Con A-induced mouse T cell activation in vitro. The inhibitory effect of Ssd on PMA-induced T cell activation was associated with down regulation of NF-kappaB signaling through suppression of IKK and Akt activities. In addition, Ssd suppressed both DNA binding activity and the nuclear translocation of NF-AT and activator protein 1 (AP-1) of the PMA/Ionomycin stimulated T cells. The cell surface markers like IL-2 receptor (CD25) were also down-regulated together with decreased production of pro-inflammatory cytokines of IL-6, TNF-alpha and IFN-gamma. These results indicate that the NF-kappaB, NF AT and AP-1 (c-Fos) signaling pathways are involved in the T cell inhibition evoked by Ssd, so it can be a potential candidate for further study in treating T cell-mediated autoimmune conditions. PMID- 19301262 TI - Identification of tubulins as substrates of serine protease HtrA1 by mixture based oriented peptide library screening. AB - Serine protease HtrA1 belongs to a family of chymotrypsin-like proteases that were first identified in bacteria and later in mammalian systems. These proteases were identified as components of protein quality control in prokaryotic systems and as regulators of diverse signaling pathways in mammalian systems. In particular, HtrA1 is implicated in trophoblast cell migration and invasion, tumor progression, chemotherapy-induced cytotoxicity, osteoarthritis, age-related macular degeneration, and pathogenesis of Alzheimer's disease. However, systematic analysis of its potential substrates in biological system is still lacking. Therefore, we performed a mixture-based oriented peptide library screening to identify putative substrates of HtrA1. We identified [AEGR]-[LAGR] [IAMLR]-[TVIAL] as consensus residues for P1 to P4 sites. We identified several putative substrates of HtrA1 involved in the pathogenesis of various diseases. In this study, we report on the identification of tubulins as potential substrates of HtrA1, and validated tubulins as in vitro and intracellular substrates of HtrA1. These results provide initial insights into substrate identification and functional characterization of HtrA1 in pathogenesis of various diseases. PMID- 19301263 TI - Design of a multiphase osteochondral scaffold III: Fabrication of layered scaffolds with continuous interfaces. AB - There is a need to improve current treatments for articular cartilage injuries. This article is the third in a series describing the design and development of an osteochondral scaffold based on collagen-glycosaminoglycan and calcium phosphate technologies for regenerative repair of articular cartilage defects. The previous articles in this series described methods for producing porous, three-dimensional mineralized collagen-GAG (CGCaP) scaffolds whose composition can be reproducibly varied to mimic the composition of subchondral bone, and pore microstructure and mineral phase can be modified. This article describes a method, "liquid-phase cosynthesis," that enables the production of porous, layered scaffolds that mimic the composition and structure of articular cartilage on one side, subchondral bone on the other side, and the continuous, gradual or "soft" interface between these tissues: the tidemark of articular joints. This design enables the layered scaffolds to be inserted into the subchondral bone at an osteochondral defect site without the need for sutures, glue, or screws, with a highly interconnected porous network throughout the entire osteochondral defect. Moreover, the differential moduli of the osseous and cartilaginous compartments enable these layered scaffolds to exhibit compressive deformation behavior that mimics the behavior observed in natural articular joints. PMID- 19301264 TI - Design of a multiphase osteochondral scaffold. I. Control of chemical composition. AB - This is the first in a series of articles that describe the design and development of a family of osteochondral scaffolds based on collagen glycosaminoglycan (collagen-GAG) and calcium phosphate technologies, engineered for the regenerative repair of defects in articular cartilage. The osteochondral scaffolds consist of two layers: a mineralized type I collagen-GAG scaffold designed to regenerate the underlying subchondral bone and a nonmineralized type II collagen-GAG scaffold designed to regenerate cartilage. The subsequent articles in this series describe the fabrication and properties of a mineralized scaffold as well as a two-layer (one mineralized, the other not) osteochondral scaffold for regeneration of the underlying bone and cartilage, respectively. This article describes a technology through which the chemical composition particularly the calcium phosphate mass fraction-of triple coprecipitated nanocomposites of collagen, glycosaminoglycan, and calcium phosphate can be accurately and reproducibly varied without the need for titrants or other additives. Here, we describe how the mineral:organic ratio can be altered over a range that includes that for articular cartilage (0 wt % mineral) and for bone (75 wt % mineral). This technology achieves the objective of mimicking the composition of two main tissue types found in articular joints, with particular emphasis on the osseous compartment of an osteochondral scaffold. Exclusion of titrants avoids the formation of potentially harmful contaminant phases during freeze-drying steps crucial for scaffold fabrication, ensuring that the potential for binding growth factors and drugs is maintained. PMID- 19301265 TI - Rapid screening, in vitro study of metal oxide and polymer hybrids as delivery coatings for improved soft-tissue integration of implants. AB - Metal-organic chemistry allows for molecular mixing and creation of a range of submicron phase-separated structures from normally brittle metal oxides and flexible polymers with improved bioactivity and delivery properties. In this study, we used a high throughput platform to investigate the influence of organic metal oxide doping of polydimethylsiloxane (PDMS) coatings on cellular bioactivity and controlled release of vanadium compared with titanium oxide coatings without additional PDMS. Metal-organic-derived titanium and or vanadium was doped into PDMS and used to form a coating on the bottom of cell culture microplates in the absence of added water, acids, or bases. These hybrid coatings were rapidly screened to establish how titanium and vanadium concentration influences cell proliferation, adhesion, and morphology. We demonstrate that titanium doping of PDMS can be used to improve cell proliferation and adhesion, and that vanadium doping caused a biphasic dose response in proliferation. A 28 day vanadium and titanium elution study indicated that titanium was not released, but the presence of PDMS in coatings increased delivery rates of vanadium compared with titania coatings without polymer. Hybrid coatings of titanium-doped polymers have potential for improving wound healing dynamics, soft-tissue integration of medical implants, and use as controlled delivery vehicles. PMID- 19301266 TI - Phenotypic changes in proliferation, differentiation, and migration of chondrocytes: 3D in vitro models for joint wound healing. AB - We aim to establish a 3D model of cartilage wound healing, and explore the involvement of chondrocytes in its repair. To characterize chondrocyte involvement in wound healing, an in vitro 3D model composed of chondrocyte mixing with either type II/I collagen or type I collagen matrix was established. The "defects" measuring 5 mm in diameter were made on each collagen matrix chondrocyte construct to mimic in vivo cartilage defects. The effects of basic fibroblast growth factor (bFGF) on chondrocytes migration and differentiation were studied. The migration and Glucosaminoglycan (GAG) synthesis of chondrocytes in the defect areas were observed by microscopy after Alcian-blue staining. In the presence of bFGF, GAG expression increased significantly when chondrocytes were cultured in type II/I collagen matrix compared to type I collagen matrix. However, mild GAG accumulation was also found when cells were cultured in either type I or type II/I collagens without bFGF. In a 3D model of cartilage wound healing, bFGF promote chondrocyte proliferation, migration and differentiation in the presence of type II/I collagen matrix, and showed potential to regulate wound healing. These wound healing models may provide feasible methods to explore various drugs prior to human trials. PMID- 19301267 TI - An immunofluorescence method for postembedded tissue in the acrylic resin Technovit 9100 New using fluorescein isothiocyanate secondary detection. AB - Immunofluorescence labeling on postembedded tissue in resin is a formidable task. Although resin components and stabilizers are a source of additional strong native fluorescence that overlaps with absorption and emission spectra of commonly used green fluorophores, the unfixed tissue is also subject to native fluorescence. For tissue embedded in resin, we hypothesized that initially removing the resin and subsequently quenching the native fluorescence from the sample could result in specific immunofluorescence signals. The hypothesis was tested on fixed tissue samples embedded in Technovit 9100 New. Deacrylated and rehydrated semithin sections from a variety of soft tissues were exposed to a quenching solution prior to immunolabeling. Cryostat sections from snap frozen tissue were also stained to assess whether all antigens investigated in fixed tissue were adequately detected. The secondary detection included antibodies conjugated with fluorescein isothiocyanate. The results were evaluated using conventional dark-field and confocal laser scanning microscopy. Both forms of microscopy confirmed the considerable lowering of the native fluorescence associated with the resin and fixed tissue samples with enhanced specific signal. The cryostat tissue sections using the same antibodies in equivalent concentrations confirmed labeling of the same cellular sites as those observed in the fixed tissue. This article describes a method for immunofluorescence labeling in Technovit 9100 New resin embedded tissue and suggests the likely chromogenic elements generating autofluorescence. PMID- 19301268 TI - Expansion of the human embryonic brain during rapid growth: area analysis. AB - This report focuses on growth of the brain of the early human embryo, Carnegie stages 12-23. Areas of median sections from 50 to 58 embryos were measured to determine the best mathematical model to describe growth of the three primary brain vesicles and to determine the change in the ratio of tissue to cavity areas (T/C). An exponential model best describes growth of the brain and head during this time period. The head expands 248-fold compared with a 171-fold growth of the brain. The whole brain, forebrain, and midbrain all exhibit larger cavities than tissue initially followed by a reversal of such at a critical time (stages 21-24). The presumptive cerebellar tissue which was twice the cavity initially grows to become more than six times the cavity. Boxplots of the T/C ratios for the head and brain plus its components reveal that initially the tissue is less than the cavity (10-20% and 40-60%, respectively) but eventually becomes larger (60-200%). PMID- 19301269 TI - Three-dimensional reconstruction of efferent ducts in wild-type and Lgr4 knock out mice. AB - We have recently shown that Lgr4 knock-out (LGR4KO) male mice are infertile due to a developmental defect of the reproductive tract. Spermatozoa do not reach the epididymis and accumulate at the rete testis and efferent ducts (ED). We have proposed that in LGR4KO, ED might fail to connect resulting in blind-ended tubes that preclude the normal transit of sperm cells. To explore this possibility, we reconstructed the three-dimensional (3D) structure of the organ from serial microphotographs. The resulting model allowed to individualize and follow each ED from the testis up to the epididymis, and to display the spatial distribution of their content. The transit of spermatozoa is indeed blocked in LGR4KO mice but, contrary to the expectation, the ducts connect normally to each other, forming a single tube that flows into the epididymis, as in the wild-type animals. In the KO however, transit of the sperm is abruptly blocked at the same level syncytial like aggregates appear in the luminal space. The model also allowed calculating, for the first time, morphometric parameters of the mouse ED, such as total volume, surface, radius, and length. These data unambiguously showed that ED in the mutant mouse are dramatically shortened and less convoluted than in the wild type animal, providing an explanation to the phenotype observed in LGR4KO. Combined with in situ immunodetection or RNA in situ hybridization, 3D reconstruction of serial histological sections will provide an efficient mean to study expression profiles in organs which do not lend themselves to whole-mount studies. PMID- 19301270 TI - Demyelination induces the decline of the myelinated fiber length in aged rat white matter. AB - To determine the exact reason for the age-related decline of the myelinated fiber length in white matter, we performed this study. In middle-aged rats, there was age-related loss of the unmyelinated fibers with large diameters. The demyelination of the myelinated fibers with small diameters in middle-aged rat white matter might make the age-related decrease of the unmyelinated fibers with small diameters in the white matter unnoticeable. However, in old-aged female rats, the unmyelinated fibers with large and small diameters significantly degenerated together and that the unmyelinated fibers formed from the demyelination of the myelinated fibers could not replenish the age-related loss of the unmyelinated fibers in the white matter. In conclusion, this study suggested that demyelination of myelinated fibers with small diameters in aged white matter might be the key mechanism of the significant decline of the myelinated fiber length in aged white matter. PMID- 19301273 TI - Porous beta tricalcium phosphate scaffolds used as a BMP-2 delivery system for bone tissue engineering. AB - Macroporous beta tricalcium phosphate (beta-TCP) scaffolds were evaluated as potential carriers and delivery systems for bone morphogenetic protein-2 (BMP-2). Chemical etching was performed to increase the available surface and thus the protein loading. X-ray diffraction and infrared spectrocopy analyses confirmed the preparation of pure beta-TCP scaffolds. Scanning electron microscopy revealed interconnected porosity (64%) and a microporous surface after chemical etching. Scaffolds loaded with 30 and 15 microg of BMP-2 were implanted respectively into the back muscles and into femoral defects (condyle and diaphysis) of rabbits for 4 weeks. Histological observations confirmed the activity of the BMP-2 released from the scaffolds. Intramuscularly, bone was formed within the BMP-2-loaded scaffold pores. In the bone defects, the effect of released BMP-2 was similarly noticeable, as evaluated by histomorphometry. The incorporation of BMP-2 resulted in an amount of newly formed bone that was 1.3 times higher than with unloaded scaffolds. The implant site, however, did not have an effect on bone formation as no statistical differences were measured between cortical (diaphysis) and trabecular (condyle) defects. These results indicate the suitability of chemically etched beta-TCP scaffolds as BMP-2 carriers, in the context of bone regeneration. PMID- 19301272 TI - Effects of tissue age on bone tissue material composition and nanomechanical properties in the rat cortex. AB - Although osteoporosis is known to alter bone tissue composition, the effects of such compositional changes on tissue material properties have not yet been examined. The natural gradient in tissue mineral content arising from skeletal appositional growth provides a basic model for investigation of relationships between tissue composition and mechanical properties. The purpose of this study was to examine the effects of tissue age on bone tissue composition and nanomechanical properties. The nanomechanical properties and composition of regions of differing tissue age were characterized in the femoral cortices of growing rats using nanoindentation and Raman spectroscopy. In addition, spatial maps of the properties of periosteal tissue were examined to investigate in detail the spatial gradients in the properties of newly formed tissue. Newly formed tissue (0-4 days) was 84% less stiff and had 79% lower mineral:matrix ratio than older intracortical (15-70 days) tissue. Tissue modulus, hardness, mineral:matrix ratio, and carbonate:phosphate ratio increased sharply with distance from the periosteum and attained the properties of intracortical tissue within 4 days of formation. The mineral: matrix ratio explained 54% and 62% of the variation in tissue indentation modulus and hardness, respectively. Our data demonstrate significant variations in tissue mechanical properties with tissue age and relate mechanical properties to composition at the microscale. PMID- 19301274 TI - Design of a multiphase osteochondral scaffold. II. Fabrication of a mineralized collagen-glycosaminoglycan scaffold. AB - This paper is the second in a series of papers describing the design and development of an osteochondral scaffold using collagen-glycosaminoglycan and calcium phosphate technologies engineered for the regenerative repair of articular cartilage defects. The previous paper described a technology (concurrent mapping) for systematic variation and control of the chemical composition of triple coprecipitated collagen, glycosaminoglycan, and calcium phosphate (CGCaP) nanocomposites without using titrants. This paper describes (1) fabricating porous, three-dimensional scaffolds from the CGCaP suspensions, (2) characterizing the microstructure and mechanical properties of such scaffolds, and (3) modifying the calcium phosphate mineral phase. The methods build on the previously demonstrated ability to vary the composition of a CGCaP suspension (calcium phosphate mass fraction between 0 and 80 wt %) and enable the production of scaffolds whose pore architecture (mean pore size: 50-1000 microm), CaP phase chemistry (brushite, octacalcium phosphate, apatite) and crosslinking density (therefore mechanical properties and degradation rate) can be independently controlled. The scaffolds described in this paper combine the desirable biochemical properties and pore architecture of porous collagen-glycosaminoglycan scaffolds with the strength and direct bone-bonding properties of calcium phosphate biomaterials in a manner that can be tailored to meet the demands of a range of applications in orthopedics and regenerative medicine. PMID- 19301275 TI - Pancreatic cancer stem cells and relevance to cancer treatments. AB - Pancreatic cancer continues to be a malignancy with few therapeutic options. The majority of patients that present for an evaluation have locally advanced or metastatic disease that is incurable by surgical approaches. Chemotherapy and radiotherapy resistance of pancreatic adenocarcinomas limits the efficacy of these therapeutic approaches. Recent evidence supports the existence of human pancreatic cancer stem cells, which appear to drive tumor initiation and progression and are particularly resistant to cell death induced by radiation or chemotherapy. Understanding the mechanisms of pancreatic cancer stem cell self renewal and resistance to standard therapies may lead to new, more effective therapies to treat this dismal disease. PMID- 19301276 TI - Endomorphin 1- and endomorphin 2-containing neurons in nucleus tractus solitarii send axons to the parabrachial nuclei in the rat. AB - Endomorphin-1 (EM1) and endomorphin-2 (EM2) are the selective endogenous ligands for the mu-opioid receptor (MOR). Since EMs-expressing neuronal cell bodies or axonal components have been observed, respectively, in the nucleus tractus solitarii or the parabrachial nuclei, we examined if EMs-expressing neurons in the NTS of the rat might send their axons to the PBN. Immunofluorescent stainings for EM1 or EM2 were combined with retrograde or anterograde tract-tracing method. After injecting tetramethyl rhodamine dextran-amine (TMR) into the parabrachial nuclei of rats, some EM1- or EM2-immunoreactive neurons in the nucleus tractus solitarii were labeled retrogradely with TMR. The majority of the EM1/TMR and EM2/TMR double-labeled neurons were observed in the medial, commissural, and dorsolateral subnuclei of the nucleus tractus solitarii. Following injection of biotinylated dextran amine (BDA) into the medial, commissural, or dorsolateral subnuclei of the nucleus tractus solitarii, EM1- or EM2-immunopositive axons and axon terminals were anterogradely labeled with BDA mainly in the lateral parabrachial nucleus. The present results have indicated that endomorphinergic neurons in the nucleus tractus solitarii project to the parabrachial nuclei. This suggests that EMs released from NTS-PBN projection fibers may bind to MOR on the PBN neurons to be implicated in processing of visceral information within the parabrachial nuclei. PMID- 19301277 TI - Construction of a multiple myeloma diagnostic model by magnetic bead-based MALDI TOF mass spectrometry of serum and pattern recognition software. AB - A diagnosis of multiple myeloma (MM) is difficult to make on the basis of any single laboratory test result. Accurate diagnosis of MM generally results from a number of costly and invasive laboratory tests and medical procedures. The aim of this work is to find a new, highly specific and sensitive method for MM diagnosis. Serum samples were tested in groups representing MM (n = 54) and non MM (n = 108). These included a subgroup of 17 plasma cell dyscrasias, a subgroup of 17 reactive plasmacytosis, 5 B cell lymphomas, and 7 other tumors with osseus metastasis, as well as 62 healthy donors as controls. Bioinformatic calculations associated with MM were performed. The decision algorithm, with a panel of three biomarkers, correctly identified 24 of 24 (100%) MM samples and 46 of 49 (93.88%) non-MM samples in the training set. During the masked test for the discriminatory model, 26 of 30 MM patients (sensitivity, 86.67%) were precisely recognized, and all 34 normal donors were successfully classified; patients with reactive plasmacytosis were also correctly classified into the non-MM group, and 11 of the other patients were incorrectly classified as MM. The results suggested that proteomic fingerprint technology combining magnetic beads with MALDI-TOF-MS has the potential for identifying individuals with MM. The biomarker classification model was suitable for preliminary assessment of MM and could potentially serve as a useful tool for MM diagnosis and differentiation diagnosis. PMID- 19301278 TI - Rotation of organizer tissue contributes to left-right asymmetry. AB - Current hypotheses regarding vertebrate left-right asymmetry patterns are based on the presumption that genetic regulatory networks specify sidedness via extracellular morphogens and/or ciliary activity. We show empirical time-lapse evidence for an asymmetric rotation of epiblastic nodal tissue in avian embryos. This rotation spans the interval when initial symmetric expression of Shh and Fgf8 becomes asymmetrical with respect to the midline. PMID- 19301279 TI - Function of sirtuins in biological tissues. AB - Sirtuins are protein deacetylases, which are dependent on nicotine adenine dinucleotide. They are phylogenetically conserved from bacteria to humans. Seven sirtuin proteins localized in a wide variety of subcellular locations have been identified in the human genome. The most important known function of sirtuins is their regulation of transcriptional repression, mediated through binding of a complex containing sirtuins and other proteins. Studies have shown that sirtuins have pathophysiological relevance to neurodegeneration, muscle differentiation, inflammation, obesity, and cancer. In addition, sirtuin activity extends the lifespan of several organisms. In this review, we discuss the mode(s) of action of sirtuins, and their biological role(s) in health and disease. PMID- 19301280 TI - Age classes and sex differences in the skull of the Mediterranean monk seal, Monachus monachus (Hermann, 1779). A study based on bone shape and density. AB - This study analyzes morphometrically 17 skulls of the Mediterranean monk seal Monachus monachus housed in different Italian Museums and collections. We considered several morphometric variables (31 linear, 1 volumetric and 1 surface area measurements). In addition, we identified, measured and compared two nonmorphometric variables, namely, the bone densities of selected areas obtained using a dual-energy X-ray absorptiometry (DXA) device. The high correlation coefficient of all variables indicated continuous growth with the onset of age. The ranking of the hierarchical cluster analysis identified the presence of three main groups containing individuals of similar sizes: lactating pups and yearlings; subadult individuals and adult females; and adult males. Smaller groups were identified within these clusters, and their respective allocations into two subgroups were argued on the basis of skull development and other factors. The discriminant analysis of the three main groups indicated a discriminant diagnostic key, based on condilobasilar length (CBlr-L); maximum mandibular branch height (MB-H); and surface area of the bulla tympanica. The proposed diagnostic key is useful to classify monk seal skulls of unidentified age and sex. The data reported here suggest that in this species certain adult skull growth features (enhanced tympanic bullae surface area extension, occipital bone density) are sexually dimorphic and possibly related to specific anatomical functions. These functions may include an enhanced auditory capacity; an increased development of the cranial musculature capable of supporting a large skull and guaranteeing the mandibular strength necessary for mastication; and male to male social interactions. PMID- 19301281 TI - The impact of dietary organic and transgenic soy on the reproductive system of female adult rat. AB - The goal of this article was to compare the effects of a prolonged use of organic and transgenic soy on the lipid profile and ovary and uterus morphology. Wistar rats were fed three different diets from weaning until sacrifice at 15 months of age. The three diets were: casein-based diet control group (CG), organic soy based diet group (OSG), or transgenic soy-based diet group (GMSG). There were no differences in food consumption or in the diet isoflavone components among the groups. Compared with the CG diet, both the OSG and GMSG diets were associated with significant reductions in body weight, serum triglycerides, and cholesterol (P < 0.05) (CG = 406 +/- 23.1; 104.3 +/- 13.2; 119.9 +/- 7.3 GMSG = 368 +/- 17.6; 60.3 +/- 4.6; 83.3 +/- 5.7 OSG = 389 +/- 23.5; 72.3 +/- 12.5; 95.5 +/- 8.0, respectively). The volume density of endometrial glandular epithelium was greater in the GMSG group (29.5 +/- 7.17, P < 0.001) when compared with the CG (18.5 +/- 7.4) and OSG (20.3 +/- 10.6) groups. The length density of endometrial glandular epithelium was shorter in both GMSG (567.6 +/- 41.1) and OSG (514.8 +/- 144.5) diets compared with the CG (P < 0.05) diet. GMSG also resulted in reduced follicle number and increased corpus luteum number compared to the OSG or CG diets (P < 0.05). In summary, both GMSG and OSG diets resulted in decreased body weight and lower serum triglyceride and cholesterol levels, and alterations in uterine and ovarian morphology were also observed. The prolonged use of soy-based diets and their relation to reproductive health warrants further investigation. PMID- 19301282 TI - Neurochemistry of olivocochlear neurons in the hamster. AB - The present study was conducted to characterize the superior olivary complex (SOC) of the lower brain stem in the pigmented Djungarian hamster Phodopus sungorus. Using Nissl-stained serial cryostat sections from fresh-frozen brains, we determined the borders of the SOC nuclei. We also identified olivocochlear (OC) neurons by retrograde neuronal tracing upon injection of Fluoro-Gold into the scala tympani. To evaluate the SOC as a putative source of neuronal nitric oxide synthase (nNOS), arginine-vasopressin (AVP), oxytocin (OT), vasoactive intestinal polypeptide (VIP), or pituitary adenylate cyclase-activating polypeptide (PACAP) that were all found in the cochlea, we conducted immunohistochemistry on sections exhibiting retrogradely labeled neurons. We did not observe AVP-, OT-, or VIP-immunoreactivity, neither in OC neurons nor in the SOC at all, revealing that cochlear AVP, OT, and VIP are of nonolivary origin. However, we found nNOS, the enzyme responsible for nitric oxide synthesis in neurons, and PACAP in neuronal perikarya of the SOC. Retrogradely labeled neurons of the lateral olivocochlear (LOC) system in the lateral superior olive did not contain PACAP and were only infrequently nNOS-immunoreactive. In contrast, some shell neurons and some of the medial OC (MOC) system exhibited immunofluorescence for either substance. Our data obtained from the dwarf hamster Phodopus sungorus confirm previous observations that a part of the LOC system is nitrergic. They further demonstrate that the medial olivocochlear system is partly nitrergic and use PACAP as neurotransmitter or modulator. PMID- 19301283 TI - Resting chest pain, negative treadmill excercise electrocardiogram, and reverse redistribution in dipyridamole myocardial perfusion scintigraphy might be the features of coronary artery spasm. PMID- 19301284 TI - Clinical diagnosis of posttraumatic stress disorder after myocardial infarction. AB - BACKGROUND: Clinician-rated large-scale studies estimating the prevalence of posttraumatic stress disorder (PTSD) related to myocardial infarction (MI) and identifying predictors of clinical PTSD are currently lacking. HYPOTHESES: We hypothesized that PTSD is prevalent in post-MI patients and that the subjective experience of the MI determines PTSD status. METHODS: We approached 951 post-MI patients with a questionnaire screening for PTSD symptoms related to their MI. Those responding and meeting a cutoff of PTSD symptom levels were invited to participate in a structured clinical interview to diagnose PTSD following Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria. Fear of dying, feelings of helplessness, and severity of pain perceived during the MI were also assessed by visual analog scales. RESULTS: The screening questionnaire was completed by 394 patients, whereby 77 met the cutoff for the interview (8 patients declined the interview). Forty of 394 patients (10.2%) had clinical PTSD (subsyndromal and syndromal forms combined). Younger age (OR 0.95, 95% CI 0.91 0.99), greater fear of dying (OR 2.77, 95% CI 1.28-5.97), and more intense feelings of helplessness (OR 2.97, 95% CI 1.42-6.21) were independent predictors of PTSD status. Perceived pain intensity during MI, sex, type of index MI, left ventricular ejection fraction, number of coronary occlusions, and highest level of total creatinine kinase were not significant predictors. CONCLUSIONS: Clinical PTSD is prevalent in post-MI patients. Demographic and particularly psychological variables related to the subjective experience of the event were stronger predictors of PTSD status than were objective measures of MI severity. PMID- 19301285 TI - Regional pericarditis: a review of the pericardial manifestations of acute myocardial infarction. AB - Regional pericarditis has been described in several settings, but occurs most frequently after transmural myocardial infarction. While the diagnosis remains elusive, it must be considered in all patients with recurrent chest pain following acute myocardial infarction (AMI). Pericarditis classically presents with positional chest pain, a pericardial friction rub, diffuse ST-segment elevation, and PR depression, but regional ECG changes associated with infarction associated pericarditis sometimes exist. Given the magnitude and frequency of AMI, it is imperative to be aware of the myriad of pericardial manifestations of myocardial injury. An illustrative case and a comprehensive review of the literature will be provided. PMID- 19301286 TI - Retraction. The role of biological extracellular matrix scaffolds in vascularized three-dimensional tissue growth in vivo. PMID- 19301287 TI - Synthesis and property of a novel calcium phosphate cement. AB - The beta-cyclodextrins (beta-CDs) and amorphous calcium phosphate were used to prepare a novel calcium phosphate cement (CPC) after mechanochemical activation. Under the mechanochemical treatment, hydrogen bonding formed between beta-CD molecules, which induced strong interaction among beta-CD molecules in aqueous solution, thus conferring good antiwashout property for CPC. After mechanochemical treatment, the solubility of beta-CD was increased and so beta-CD showed different behaviors within and on the outer surface of CPC. During the setting process, when beta-CD was injected it flowed to the surface of CPC, forming a beta-CD film on the surface of CPC; inside the cement, the hydration reaction of calcium phosphates was not affected. Moreover, the antiwashout CPC with beta-CD also showed good mechanical setting, rheological properties, and injectability. The novel CPC was supposed to be developed as a good injectable bone substitute material in the clinical application. PMID- 19301288 TI - Use of emergency medical services in patients with acute myocardial infarction in China. AB - BACKGROUND: Although guidelines strongly recommend use of the Emergency Medical Systems (EMS) by patients with acute myocardial infarction (AMI), it remains underutilized in western countries. Information about its current use in China is unclear. The objective of this study was to examine the use of the EMS by patients with AMI in China, and investigate factors affecting its use. METHODS: A prospective survey study, which included 803 patients with AMI who were admitted to 21 hospitals in China between November 1, 2005 and December 31, 2006. RESULTS: Only 39.5% of patients called up the EMS at the onset of symptoms (EMS group, n=317), whereas the rest presented to the hospital by some other means (self transport group, n=486, 60.5%). Predictors of EMS users were older age, symptom onset at evening, unbearable symptoms, having received training and acquired knowledge on heart attack, as well as having a higher income and medical history of heart failure or stroke. Prehospital delay (median 110 min vs. 143 min, p<0.001), door to needle time (median 85 min vs. 93 min, p<0.005) and door-to balloon time (median 118 min vs. 160 min, p<0.001) were significantly shorter in the EMS group. The early reperfusion rate was also significantly higher in the EMS group (84.8% vs. 78.2%, p=0.019), mainly because of a greater incidence of primary percutaneous coronary intervention (68.1% vs. 61.7%, p=0.046). CONCLUSIONS: The emergency medical services are underutilized by patients with AMI in China. Use of the EMS may be advantageous in view of greater administration of reperfusion therapy. New public health strategies should be developed to facilitate greater use of the EMS for AMI. PMID- 19301289 TI - The impact of aspirin resistance on the long-term cardiovascular mortality in patients with non-ST segment elevation acute coronary syndromes. AB - BACKGROUND: Aspirin resistance has been associated with an adverse long-term outcome in patients with atherosclerotic coronary artery disease, but more studies are needed. HYPOTHESIS: The aim of this study was to investigate the impact of aspirin resistance, assessed by the Platelet Function Analyzer-100 (PFA 100) (Dade Behring Inc., Deerfield, Ill., USA) on the long-term prognosis in patients with non-ST segment elevation acute coronary syndromes (NSTE-ACS). METHODS: A total of 496 consecutive patients were studied. The 1-y incidence of cardiovascular death was the prespecified study endpoint. The patients were divided, according to the values of PFA-100 collagen epinephrine closure time (CEPI-CT) upon presentation, into aspirin sensitives (those with a PFA-100 CEPI CT>193 sec) and aspirin resistants (those with a PFA-100 CEPI-CTor=50%; 77.0+/-4.9 versus 14.0+/-50.9, p=0.017), but not BNP.However, BNP levels in pericardial fluid were significantly higher in patients with left ventricle dilatation (left ventricular diastolic dimension54 mm; 130.3+/-68.9: 709.7+/ 324.7, p=0.0168). Moreover, BNP levels in pericardial fluid were significantly higher in Grade III than Grade II and I (Grade I: echo-free space<5 mm, Grade II: 5-10 mm, Grade III: >10 mm). CONCLUSION: These results suggest that BNP levels in pericardial fluid served as more sensitive and accurate indicators of left ventricular diastolic dysfunction, and that increased BNP levels in pericardial fluid may have an important pathophysiologic role in heart failure as a cardiocyte-derived antifibrotic factor. PMID- 19301293 TI - Prolonged dual antiplatelet therapy improves clinical outcomes in high-risk patients implanted with sirolimus-eluting stents. AB - BACKGROUND: Although a science advisory recommending 12 months of dual antiplatelet therapy after drug-eluting stents implantation was published recently, the optimal duration of dual antiplatelet therapy has not yet been precisely determined. HYPOTHESIS: Prolonged dual antiplatelet therapy can improve clinical outcomes in high-risk patients implanted with sirolimus-eluting stents. METHODS: The patients implanted with sirolimus-eluting stents were assigned into standard clopidogrel therapy group (clopidogrel 75 mg/d for 12 mo) and prolonged clopidogrel therapy group (clopidogrel 75 mg/d for 18 mo). Long-term aspirin (100 mg/d) therapy was adopted in both groups. The primary endpoint was very late stent thrombosis. RESULTS: After 12 months, 24 patients were excluded because of major adverse cardiovascular events (MACEs). Three hundred and thirty six patients surviving without MACEs were further followed up for 6 months. Between 12 and 18 months, in 160 patients with standard clopidogrel therapy, 5.6% had very late stent thrombosis. In contrast, in 176 patients with prolonged clopidogrel therapy, 1.1% had very late stent thrombosis (p<0.01, versus standard clopidogrel therapy group). CONCLUSIONS: Prolonged dual antiplatelet therapy may be beneficial to prevent very late stent thrombosis after sirolimus-eluting stents implantation in high-risk patients. PMID- 19301294 TI - Clinical efficiency and safety analysis of transcatheter closure of multiple atrial septal defects in adults. AB - BACKGROUND: Transcatheter closure of atrial septal defects (ASDs) is currently a reliable alternative to surgery, even though challenging in patients with multiple ASDs. HYPOTHESIS: The aim of this study was to evaluate the clinical efficiency and safety of transcatheter closure in multiple ASDs. METHODS: Multiple ASDs were diagnosed by transthoracic echocardiography (TTE) or transesophageal echocardiography (TEE). The occlusive condition and distance between 2 adjacent ASDs were measured by TTE examination. Then, the number and size of the occluder(s) was determined. TTE examinations were performed after transcatheter closure as follow-up. RESULTS: The transcatheter procedure was successful in 15 patients with multiple ASDs, using a single occluder in 9 patients and 2 occluders in the remaining 6 patients. Overall, 21 ASD occluders were implanted. During a follow-up period of 6 mo to 5 y, a slight residual shunt was found in 1 patient without any symptoms; a moderate residual shunt was identified at the inferior vena cava and the occluder was removed by surgery 1 mo after procedure. Other complications, including endocarditis, arrhythmia, thromboembolism, and atrioventricular valve damage were not recorded in any of the 15 patients during the follow-up period. CONCLUSION: Transcatheter closure of multiple ASDs is safe and efficient. Two occluders are necessary for the distance of 2 ASDs more than 7 mm, and a single occluder is sufficient for those 7 mm or less. PMID- 19301295 TI - Cardiorespiratory fitness and sedentary lifestyle in the morbidly obese. AB - BACKGROUND: Sedentary lifestyles and poor physical fitness are major contributors to the current obesity and cardiovascular disease pandemic. HYPOTHESIS: Daily physical activity and cardiorespiratory fitness are correlated in morbidly obese individuals in their free-living environment. METHODS: Ten morbidly obese participants continuously wore an activity sensor that measured caloric expenditure, minute-by-minute physical activity, and steps/day over a 72-h period. Following collection of the device data, structured cardiorespiratory fitness testing was performed on each subject. RESULTS: Mean caloric expenditure for all individuals was 2,668+/-481 kcal/d. On average, subjects took 3,763+/ 2,223 steps. On average 23 h and 51.6 min per d were spent sleeping or engaged in sedentary activity (<3 metabolic equivalents [METs]) and the remaining 8.4 min were spent in moderate activity (3-6 METs). Average peak VO2 was 16.8+/-4.7 mL/kg/min. Higher peak VO2 correlated with higher total caloric expenditure (TCE; r=0.628, p=0.05) and trended with higher steps/day (r=0.591, p=0.07). CONCLUSIONS: Most morbidly obese participants in this study were markedly sedentary. These study results may provide important links between obesity, poor fitness, and cardiovascular disease. PMID- 19301296 TI - PCI, stents, clopidogrel and non-cardiac surgery. AB - What follows in this discussion is partially abstracted from a wonderful ACCEL interview with Peter Berger by Freek W.A. Verheugt on the subject of stopping antiplatelet therapy prior to non-cardiac surgery. 1 I suspect what Dr. Berger believes is representative of the opinion of many interventional cardiologists. PMID- 19301297 TI - Exploring the interactions of gliadins with model membranes: effect of confined geometry and interfaces. AB - Mechanisms leading to the assembly of wheat storage proteins into proteins bodies within the endoplasmic reticulum (ER) of endosperm cells are unresolved today. In this work, physical chemistry parameters which could be involved in these processes were explored. To model the confined environment of proteins within the ER, the dynamic behavior of gamma-gliadins inserted inside lyotropic lamellar phases was studied using FRAP experiments. The evolution of the diffusion coefficient as a function of the lamellar periodicity enabled to propose the hypothesis of an interaction between gamma-gliadins and membranes. This interaction was further studied with the help of phospholipid Langmuir monolayers. gamma- and omega-gliadins were injected under DMPC and DMPG monolayers and the two-dimensional (2D) systems were studied by Brewster angle microscopy (BAM), polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS), and surface tension measurements. Results showed that both gliadins adsorbed under phospholipid monolayers, considered as biological membrane models, and formed micrometer-sized domains at equilibrium. However, their thicknesses, probed by reflectance measurements, were different: omega gliadins aggregates displayed a constant thickness, consistent with a monolayer, while the thickness of gamma-gliadins aggregates increased with the quantity of protein injected. These different behaviors could find some explanations in the difference of aminoacid sequence distribution: an alternate repeated - unrepeated domain within gamma-gliadin sequence, while one unique repeated domain was present within omega-gliadin sequence. All these findings enabled to propose a model of gliadins self-assembly via a membrane interface and to highlight the predominant role of wheat prolamin repeated domain in the membrane interaction. In the biological context, these results would mean that the repeated domain could be considered as an anchor for the interaction with the ER membrane and a nucleus point for the formation and growth of protein bodies within endosperm cells. (c) 2009 Wiley Periodicals, Inc. Biopolymers 91: 610-622, 2009.This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com. PMID- 19301298 TI - Folic acid, methylation and neural tube closure in humans. AB - This review provides a brief description of folate use and folic acid metabolism in relation to neural tube defect (NTD) risk. First, a meta-analysis of reduction in NTD recurrence and occurrence risk with periconceptional folic acid supplementation is presented. Second, an overview of the complex folate metabolism is given. Third, SNPs for genes involved in folate and homocysteine metabolism that have been studied in relation to NTD riskare discussed. Fourth, the questions whether folate receptor autoantibodies or hampered methylation are mechanisms underlying NTDs are briefly discussed. PMID- 19301299 TI - Control--the Striga conundrum. AB - There is a wide range of existing and potential control options for Striga. This paper describes and discusses many of the control options, with a focus on technology limitations, adoption limitations (real or potential) and, in the case of novel technologies, development limitations. The paper addresses the question as to why, after many years of research, control method testing, piloting and technology dissemination, the wide-scale effective control of Striga hermonthica (Del.) Benth. and Striga asiatica (L.) Kuntze is so elusive. Limitations, including variable technology reliability, poor access to control technology, costs (monetary, labour, skills) associated with control technology, limited practicality of methods and poor information, all hamper the adoption and impact of existing control methods. Some of the same issues may impact upon novel control technologies, and this needs careful consideration. Additional issues surround other potential technologies, especially so in the case of transgenic approaches. Suggestions are made as to how the impasse of effective Striga control can be overcome. More effective use of integrated control approaches, improved crop germplasm phenotyping, enhanced understanding of the host/non-host- parasite interaction and better integration and communication among the parasitic plant research, development and extension community are among the suggestions made. PMID- 19301300 TI - CAGE unsound for alcohol misuse in seniors. PMID- 19301301 TI - Gonadotropin releasing hormone analogs induce apoptosis by extrinsic pathway involving p53 phosphorylation in primary cell cultures of human prostatic adenocarcinomas. AB - BACKGROUND: Gonadotropin-releasing-hormone (GnRH) analogs are widely used to block hypothalamic-pituitary-gonadal axis and inhibit blood androgen levels in patients with prostate cancer (PCa). In addition, GnRH analogs induce proliferation arrest and apoptosis through GnRH receptors expressed on the membrane of PCa cells. Possible molecular mechanisms involved in GnRH-mediated apoptosis on prostate cancer cells were studied. METHODS: Primary cultures from PCa and benign prostatic hyperplasia (BPH) (non-malignant control) were derived from samples provided by our Institutional Hospital. Cell cultures were incubated for 24 hr with 20 ng/ml of GnRH agonist Leuprolide (Lp) or antagonist Cetrorelix (Cx). Apoptosis was evaluated by studying the expression of Bax and Bcl-2 and the activation of caspase-9 (intrinsic pathway), caspase-8 (extrinsic pathway), and caspase-3. Also, mRNA level, protein expression and phosphorylation of p53 were studied. RESULTS: Cleaved caspase-8 and -3, but not -9, increased in presence of Lp and Cx in PCa cell cultures. Bax and Bcl-2 mRNA levels showed no changes after GnRH-analog treatments. Only Bax protein showed an increase after Cx treatment in PCa cell cultures. p53 mRNA level was higher in PCa than in BPH cell cultures. Lp and Cx increased p53 expression and phosphorylation in PCa cell cultures. CONCLUSIONS: Apoptosis induced by GnRH analogs seems to be mediated by extrinsic pathway involving p53 phosphorylation. Phosphorylated-p53 might be associated with the increase in apoptotic NGF receptor, p75, previously reported by our laboratory. These findings reinforce the concept of clinical use of GnRH analogs for PCa suggesting that intraprostatic treatment may be more effective. PMID- 19301303 TI - Opera-hallucinosis. PMID- 19301304 TI - Tumoristatic effects of endostatin in prostate cancer is dependent on androgen receptor status. AB - BACKGROUND: Although anti-angiogenic therapy is a promising new line of therapy for prostate cancer, we recently reported that stable expression of endostatin arrested the progression of prostate cancer to poorly differentiated state and distant metastasis in TRAMP mice. However, the same therapy failed to provide any benefit when given either during or after the onset of metastatic switch. The present study determined the possible mechanisms behind the selective advantage of endostatin therapy in early-stage disease. METHODS: Angiogenesis-related gene expression analysis was performed to identify target genes and molecular pathways involved in the therapy effects. Based on the results from in vivo studies, and recapitulation of the in vivo data in vitro using tumorigenic and non-tumorigenic human prostate cancer cells that are either androgen-sensitive or androgen independent, analyses of possible mechanisms of the selective advantage of early treatment were performed using assays for cell proliferation, apoptosis, migration, and cell signaling. The identified mechanisms were further confirmed in vivo. RESULTS: Results indicated that cells with high androgen receptor (AR) expression were more sensitive to endostatin treatment than androgen-independent cells with low or no AR expression. Endostatin was found to significantly downregulate the expression of growth factors, receptor tyrosine kinases, proteases, and AR both in vitro and in vivo only when the cells express high levels of AR. Cell proliferation was not influenced by endostatin treatment but migration was significantly affected only in androgen-sensitive cells. Targeted downregulation of AR prior to endostatin treatment in androgen-sensitive cells and overexpression of AR in androgen-independent cells indicated that the effect of endostatin via AR downregulation is mediated by a non-genotropic mechanism on Ras and RhoA pathways, and independently of AR on MAPK/ERK pathway. CONCLUSIONS: These data indicate that systemically stable endostatin expression delays the onset of metastatic switch by acting on multiple pathways involving AR. PMID- 19301305 TI - Effects of simvastatin, acetylsalicylic acid, and rosiglitazone on proliferation of normal and cancerous prostate epithelial cells at therapeutic concentrations. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs and cholesterol-lowering statins have been reported to inhibit prostate cancer cell growth suggesting their chemopreventive potential within the prostate. However, the effect has been demonstrated only with advanced prostate cancer cell lines and with drug concentrations above the clinical therapeutic range. In this study we compared the effect of therapeutic concentrations of acetylsalicylic acid, simvastatin and rosiglitazone on the growth of a set of prostatic primary cultures and various prostate epithelial cell lines. METHODS: Two primary epithelial cell lines isolated from surgical resecates of normal prostate tissue (P96E, P97E), a primary cell line isolated from untreated prostate carcinoma (ESTO1), two transformed prostate epithelial cell lines (PWR1-E, RWPE-1) and advanced cancer cell lines LNCaP and VCaP were used in the study. Cells were treated for seven days with therapeutic concentrations of acetylsalisylic acid, simvastatin, rosiglitazone or their combination. Cellular growth rate was measured by crystal violet staining method. RESULTS: Acetylsalicylic acid (0.5 mM) and simvastatin (10 nM) inhibited the growth of prostate epithelial cells of normal and primary cancer origin, whereas advanced cancer cell lines were resistant to the effect. Rosiglitazone at the therapeutic level of 1 microM did not reduce the growth of any cell type studied. CONCLUSIONS: Our results demonstrate that acetylsalicylic acid and simvastatin inhibit prostate epithelial cell growth at clinically relevant doses. This should be acknowledged when designing possible prostate cancer chemopreventive trials. PMID- 19301306 TI - Are mixed explicit/implicit solvation models reliable for studying phosphate hydrolysis? A comparative study of continuum, explicit and mixed solvation models. AB - Phosphate hydrolysis is ubiquitous in biology. However, despite intensive research on this class of reactions, the precise nature of the reaction mechanism remains controversial. Herein, we have examined the hydrolysis of three homologous phosphate diesters. The solvation free energy was simulated by means of either an implicit solvation model (COSMO), hybrid quantum mechanical/molecular mechanical free energy perturbation (QM/MM-FEP) or a mixed solvation model in which N water molecules were explicitly included in the ab initio description of the reacting system (where N=1-3), with the remainder of the solvent being implicitly modelled as a continuum. Here, both COSMO and QM/MM FEP reproduce DeltaG(obs) within an error of about 1 kcal mol(-1). However, we demonstrate that in order to obtain any kind of reliable results from a mixed model, it is essential to carefully select the explicit water molecules from short QM/MM runs that act as a model for the true infinite system. Additionally, the mixed models tend to be increasingly unstable and miss larger entropic contributions as more explicit water molecules are placed into the system. Thus, our analysis indicates that this approach provides an unreliable way for modelling phosphate hydrolysis in solution. PMID- 19301307 TI - Spectroscopic investigation of the ultrafast photoinduced dynamics in pi conjugated terpyridines. AB - Ultrafast light-induced processes in a series of pi-conjugated mono-, bis-, tris- and tetrakis(terpyridine) derivatives are investigated by femtosecond time resolved spectroscopy. Non-exponential excited-state dynamics involving singlet triplet intersystem crossing are observed which span from picoseconds to nanoseconds (see figure). Time-resolved spectroscopy is applied to investigate the ultrafast relaxation dynamics of several pi-conjugated mono-, bis-, tris- and tetrakis(terpyridine) derivatives. This particular series of structurally closely related systems was prepared applying efficient synthetic strategies and resembles key building blocks for a wide range of photoactive complexes, dendrimers and metallo-polymers with resulting potential applications, for example, in photovoltaics or as organic light-emitting diodes. Aiming for applications of supramolecular assemblies based on these recently presented terpyridine ligands a detailed knowledge of the light-induced processes of the ligands themselves represents a prerequisite. By applying femtosecond time resolved absorption spectroscopy in concert with time-resolved fluorescence and Raman measurements, we detail the photophysical properties. PMID- 19301308 TI - The excited-state lifetimes in a G x C DNA duplex are nearly independent of helix conformation and base-pairing motif. AB - DNA photophysics: Femtosecond transient absorption experiments reveal that excited states produced by UV light in a duplex DNA oligonucleotide decay at essentially the same rate in B and Z helix conformers (see figure). PMID- 19301309 TI - Regulation of cell survival by resveratrol involves inhibition of NF kappa B regulated gene expression in prostate cancer cells. AB - BACKGROUND: Polyphenols have been proposed as antitumoral agents. We have shown that resveratrol (RES) induced cell cycle arrest and promoted apoptosis in prostate cancer cells by inhibition of the PI3K pathway. The RES effects on NF kappaB activity in LNCaP cells (inducible NF kappaB), and PC-3 cells (constitutive NF kappaB) are reported. METHODS: Cells were treated with 1-150 microM of RES during 36 hr. NF kappaB subcellular localization was analyzed by western blot and immunofluorescence. I kappaB alpha was evaluated by immunoprecipitation followed by Western blot. Specific DNA binding of NF kappaB was determined by EMSA assays and NF kappaB-mediated transcriptional activity by transient transfection with a luciferase gene reporter system. RESULTS: RES induced a dose-dependent cytoplasmic retention of NF kappaB mediated by I kappaB alpha in PC-3 cells but not in LNCaP. RES-induced inhibition of NF kappaB specific binding to DNA was more significant in PC-3 cells. NF kappaB-mediated transcriptional activity induced by EGF and TNFalpha were inhibited by RES in both cell lines. LY294002 mimicked RES effects on NF kappaB activity. CONCLUSION: Antiproliferative and apoptotic effects of RES on human prostate cancer cells may be mediated by the inhibition of NF kappaB activity. This mechanism seems to be associated to RES-induced PI3K inhibition. RES could have therapeutic potential for prostate cancer treatment. PMID- 19301310 TI - Role of the VEGFR3/VEGFD receptor axis in TGFbeta1 activation of primary prostate cell lines. AB - BACKGROUND: Reports indicate that vascular endothelial growth factor receptor type 3 (VEGFR3) regulates cellular functions such as invasion, proliferation, and chemo-resistance. However, the exact function of the VEGFR3 signaling axis in prostate epithelial cells is poorly characterized. METHODS: The goal of this study was to evaluate whether TGFbeta1 in combination with VEGFD can promote pre malignant invasive activities of intermediate basal cells (IBC-10a) isolated from human prostate cancer (Gleason score 6). RESULTS: hTERT immortalized IBC-10a cells normally grew as confluent "cobblestoned" monolayers, but treatment with TGFbeta1 (10 ng/ml for 2-6 hr) dissociated the cell-cell junctions and induced VEGFR3 translocation to the cell surface. This event was not inhibited by 10 microM cycloheximide or puromycin, indicating transcription and protein synthesis were not required. We further discovered that TGFbeta1 in combination with VEGFD induced a significant increase in the invasive activity of IBC-10a cells (>26% and 53% after 24 and 48 hr, respectively) in modified Boyden Chamber assays. TGFbetaRII receptor antibodies specifically blocked TGFbeta1 induction of VEGFR3 translocation to the cell surface and blocked VEGFD-induced invasion. Zymograms revealed that TGFbeta1 (and not VEGFR3) stimulated the secretion of MMP-2 and MMP 9, presumably to promote cell invasion. The cell invasion assays confirmed that antibodies specific for TGFbetaII receptor, MMP-2 and MMP-9 and VEGFR3, independently blocked TGFbeta1-induced invasion. CONCLUSIONS: For the first time, we have demonstrated the mechanism by which TGFbeta1 stimulates VEGFD/VEGFR3 receptor axis activation leading to increased cell migration and invasion by primary intermediate basal cell cultures. PMID- 19301311 TI - A novel image-guided surgery of hepatocellular carcinoma by indocyanine green fluorescence imaging navigation. AB - BACKGROUND AND OBJECTIVES: The clear delineation between tumor and normal tissue is ideal for real-time surgical navigation imaging. We present a novel indocyanine green (ICG) fluorescence imaging technique to visualize hepatocellular carcinoma (HCC). METHODS: Ten patients with solitary HCC underwent hepatectomy between February and September 2007 at Osaka Medical Center for Cancer and Cardiovascular Diseases. ICG had been injected intravenously several days before surgery at a dose of 0.5 mg/kg body weight. After laparotomy, the liver was inspected with intraoperative ultrasonography (IOUS), and then with a near-infrared (NIR) fluorescence imaging system (PDE; Hamamatsu Photonics K.K. Hamamatsu, Japan). RESULTS: All the 10 primary tumors showed bright fluorescent signals and could be completely removed with negative margins under the guide of PDE. In four cases (40.0%), new HCC nodules that were not detected by use of any preoperative examinations including IOUS were detected by PDE. These newly identified HCC nodules were very small in size and most of the tumors were well differentiated HCCs. CONCLUSIONS: This novel technique is simple and safe, and is therefore considered to be a promising tool for routine intraoperative imaging during a hepatic resection and further clinical exploration for HCC. PMID- 19301312 TI - The puzzle of RNAs that target gene promoters. AB - Setting the right target: Most researchers who use small RNAs in mammalian cells assume that mRNA will be the target. Recent studies suggest that small RNAs can also target chromosomal DNA. Few discoveries have had the impact of RNAi. Most researchers who use small RNAs to control gene expression in mammalian cells assume that mRNA will be the target. Recent studies, however, have suggested that small RNAs can also target chromosomal DNA. PMID- 19301313 TI - Structure-based optimization of aldose reductase inhibitors originating from virtual screening. AB - Diabetes mellitus is a universal health problem. The World Health Organization (WHO) estimates that 150 million people suffer from diabetes mellitus worldwide in 2005. Long-term complications are a serious problem in the treatment of diabetes, manifesting in macrovascular and microvascular complications. Sorbitol accumulation has been proposed to be an important factor in the development of microvascular complications such as nephropathy, neuropathy, retinopathy or cataract. Catalyzing the NADPH-dependent reduction of glucose to sorbitol, aldose reductase (ALR2) is an important target in the prevention of these complications. The development of novel aldose reductase inhibitors is expected to benefit strongly from a structure-based design approach. A virtual screening based on the ultrahigh-resolution crystal structure of the inhibitor IDD 594 in complex with human ALR2 identified two compounds with IC(50) values in the low micro- to submicromolar range. Based on the known interactions between the ligands and their binding pocket, we simplified the lead structures to give the minimal structural requirements and developed synthetic pathways from commercially available compounds. The newly synthesized compounds were assayed for their inhibition of ALR2, showing inhibitory activities down to the nanomolar range. Crystal structure analysis of the most potent derivative of our series revealed insights into the binding mode of the inhibitors. PMID- 19301314 TI - Position-specific incorporation of fluorescent non-natural amino acids into maltose-binding protein for detection of ligand binding by FRET and fluorescence quenching. AB - Position-specific incorporation of fluorescent groups is a useful method for analysis of the functions and structures of proteins. We have developed a method for the incorporation of visible-wavelength-fluorescent non-natural amino acids into proteins in a cell-free translation system. Using this technique, we introduced one or two BODIPY-linked amino acids into maltose-binding protein (MBP) to obtain MBP derivatives showing ligand-dependent changes in fluorescence intensity or intensity ratio. BODIPY-FL-aminophenylalanine was incorporated in place of 15 tyrosines, as well as the N-terminal Lys1, and the C-terminal Lys370 of MBP. Fluorescence measurements revealed that MBP containing a BODIPY-FL moiety in place of Tyr210 showed a 13-fold increase in fluorescence upon binding of maltose. Tryptophan-to-phenylalanine substitutions suggest that the increase in fluorescence was the result of a decrease in the quenching of BODIPY-FL by tryptophan located around the binding site. MBP containing a BODIPY-558 moiety also showed a maltose-dependent increase in fluorescence. BODIPY-FL was then additionally incorporated in place of Lys1 of the BODIPY-558-containing MBP as a response to the amber codon. Fluorescence measurements with excitation of BODIPY FL showed a large change in fluorescence intensity ratio (0.13 to 1.25) upon binding of maltose; this change can be attributed to fluorescence resonance energy transfer (FRET) and maltose-dependent quenching of BODIPY-558. These results demonstrate the usefulness of the position-specific incorporation of fluorescent amino acids in the fluorescence-based detection of protein functions. PMID- 19301315 TI - Analysis of the indanomycin biosynthetic gene cluster from Streptomyces antibioticus NRRL 8167. AB - Metabolites that harbor a core indane scaffold are found to have diverse biological properties. Indanomycin and related pyrroloketoindanes are ionophores and have demonstrated antiparasitic, insecticidal, and antibacterial activities. To understand the biochemical mechanisms guiding formation of the central indane ring, the biosynthetic gene cluster for indanomycin was identified from Streptomyces antibioticus NRRL 8167 and sequenced to approximately 80 kb; this revealed five genes encoding subunits of a polyketide synthase (PKS) and 18 other open reading frames. The involvement of this cluster in indanomycin biosynthesis was confirmed by deletion mutagenesis. The indanomycin PKS lacks the expected thioesterase at the carboxy terminus of the final module, and instead appears to house an incomplete module containing an unusual cyclase domain. These findings now enable additional detailed genetic and biochemical studies of the mechanisms guiding the generation of pyrroloketoindanes. PMID- 19301316 TI - Fine-tuning of catalytic properties of catechol 1,2-dioxygenase by active site tailoring. AB - Catechol 1,2-dioxygenases and chlorocatechol dioxygenases are Fe(III)-dependent enzymes that do not require a reductant to perform the ortho cleavage of the aromatic ring. The reaction mechanism is common to the two enzymes, and active site residues must play a key role in the fine-tuning of specificity. Protein engineering was applied for the first time to the catalytic pocket of a catechol 1,2-dioxygenase by site-specific and site-saturation mutagenesis with the purpose of redesigning the pocket shape for improved catalysis on bulky derivatives. Mutants were analysed for changes in kinetic parameters: variants for residue 69 show an inversion of specificity with a preference towards 4-chlorocatechol (decrease of K(M) by a factor of 20) and activity on the rarely recognised substrate 4,5-dichlorocatechol, thus creating a novel, engineered chlorocatechol dioxygenase. A L69A substitution conveys gain-of-function activity towards 4-tert butylcatechol. Mutations of position 72 enhance k(cat) towards chlorinated substrates. The biphasic Arrhenius plot observed in A72S suggests the involvement of a dynamic switch in the fine regulation of the enzyme. PMID- 19301317 TI - Design, synthesis, and evaluation of carnosine derivatives as selective and efficient sequestering agents of cytotoxic reactive carbonyl species. AB - Carnosine aryl derivatives as sequestering agents of RCS: Reactive carbonyl species (RCS) are cytotoxic mediators representing a novel drug target, as they are presumed to play a pathogenic role in several diseases. Carnosine is a selective RCS-sequestering agent, but is rapidly hydrolyzed by serum carnosinase. Herein we describe the in silico design, synthesis, and evaluation of a set of carnosine aryl derivatives.Reactive carbonyl species (RCS) are important cytotoxic mediators generated by lipid oxidation of polyunsaturated fatty acids (PUFAs) and represent a novel drug target, as they are presumed to play a pathogenic role in several diseases. L-Carnosine (L-CAR, beta-alanyl-L-histidine) is a specific detoxifying agent of RCS, but is rapidly hydrolyzed in human serum by carnosinase, a specific dipeptidase. Herein we describe the in silico design, synthesis, and biological evaluation of carnosine derivatives that are resistant to carnosinase and that have increased quenching efficacy. Stability against carnosinase-mediated turnover was achieved by isomerization of the histidine residue, leading to D-carnosine (D-CAR, beta-alanyl-D-histidine), which maintains the same quenching activity of L-carnosine. A molecular modeling approach was then used to design derivatives characterized by an increased quenching efficacy. The most promising candidates were synthesized, and their stability and quenching activity were evaluated. This study describes a set of aryl derivatives that are characterized by high stability in human plasma and a quenching activity toward 4 hydroxy-trans-2-nonenal (HNE), chosen as a model of RCS, up to threefold greater than D-carnosine. PMID- 19301318 TI - Mechanistic insights into cyclooxygenase irreversible inactivation by aspirin. AB - A mechanistic hypothesis for the acetylation of cyclooxygenase (COX) by aspirin is proposed on the basis of a QM/MM study. This mechanism is consistent with previous experimental findings by other investigators. Ser 530 appears to be acetylated under intramolecular general base catalysis provided by the carboxylate moiety of aspirin, while Tyr 385 plays a crucial role in orienting and polarizing the acetyl group. PMID- 19301319 TI - Fragment-based identification of Hsp90 inhibitors. AB - Heat shock protein 90 (Hsp90) plays a key role in stress response and protection of the cell against the effects of mutation. Herein we report the identification of an Hsp90 inhibitor identified by fragment screening using a high-concentration biochemical assay, as well as its optimisation by in silico searching coupled with a structure-based drug design (SBDD) approach. PMID- 19301320 TI - Organic solar cells with semitransparent metal back contacts for power window applications. AB - To provide truly transparent solar cells for power window applications, both semiconductor and electrode materials have to have a very low absorption over as much of the visible spectrum as possible. We present some promising visible transparent semiconductor combinations, namely zinc-phthalocyanine or zinc naphthalocyanine together with soluble fullerenes in conjunction with a method for obtaining highly transparent thin metal films by tuning the interference patterns in the multilayer organic solar cells structure. In an optimal combination, solar cells with an efficiency of about 0.5 % and a peak transparency of more than 60 % in the visible part of the spectrum were fabricated. PMID- 19301321 TI - Phosphopeptide enrichment using microscale titanium dioxide solid phase extraction. AB - Identification of phosphopeptides by MS is challenging due to their relatively low abundance in proteomic samples and their limited ionization efficiency. Various affinity enrichment methods have been used in the literature. Titanium dioxide SPE devices have been recently proposed as an alternative to immobilized metal affinity chromatography for phosphopeptide enrichment. This study evaluates the TiO(2 )method using sorbent packed in a 96 well microscale extraction plate operated using a vacuum manifold. The phosphopeptide recovery and enrichment selectivity were investigated at various loading conditions. The effectiveness of organic additives such as dihydroxybenzoic acid derivatives and other nonaliphatic carboxylic acids on enrichment selectivity was examined. The performance of TiO(2) was compared to IMAC sorbent. The results suggest that various additives improve the enrichment selectivity by effectively interfering with the acidic peptides binding to TiO(2) sorbent. Interaction of phosphopeptides with sorbent is also affected, which leads to overall reduction in phosphopeptide recovery. The new SPE device was successfully utilized for the extraction of phosphopeptides from yeast lysate digest using 2,5-dihydroxybenzoic acid to minimize the interference from nonphosphorylated peptides. PMID- 19301322 TI - Effect of separation dimensions on resolution and throughput using very narrow range IEF for 2-DE after solution phase isoelectric fractionation of a complex proteome. AB - This paper describes how the lengths of the first and second dimensions in narrow pH-range 2-DE affect the number of detected protein spots, by analysis of human breast carcinoma cell line lysates prefractionated by solution phase IEF. The aim is to maximize throughput while minimizing experimental costs. In this study, systematic evaluation of narrow-range IPG strip lengths showed that separation distances were very important, with dramatic increases in resolution when longer gels were used. Compared with 7 cm minigels, maximal resolution was obtained using 18 and 24 cm IPG strips. Systematic evaluation of SDS-PAGE gel length showed a far weaker influence of separation length on resolution in the second dimension compared with that observed for the IEF dimension. There was little benefit in using separation distances greater than 12-15 cm, at least with currently available electrophoresis units. The work shows that regions of the IPG strip not containing any proteins can be excised to fit a smaller gel if prefractionation using IEF in solution has been performed. As expected, larger 2 DE gel volumes resulting from the use of longer IPG strips and second dimension gels decreased detection sensitivity when equal protein loads were used. However, this effect could be readily eliminated by increasing the loads applied to IPG strips. PMID- 19301323 TI - Shotgun mass spectrometry analysis of the human thalamus proteome. AB - The thalamus plays pivotal roles in the central nervous system as relay center for organizing information, such as auditory and visual senses from diverse brain regions and their re-distribution to the cerebral cortex. Brain diseases including schizophrenia, Parkinson's disease, epilepsy, and bipolar disorder have been associated with the thalamus. We performed a shotgun proteome analysis of iTRAQ-labeled tryptic peptides of human mediodorsal thalamus protein extracts coming from two healthy male and two healthy female subjects. The shotgun workflow consisted of IEF fractionation, RP LC and MALDI-TOF/TOF mass spectrometric analysis. We were able to identify 542 proteins that are involved in different biological processes and from diverse cellular localizations. A considerable fraction of these proteins had not been identified by traditional proteomics methods such as 2-DE. The thalamus proteome contributes to the knowledge of the human brain proteome and future applications in basic and clinical research. PMID- 19301324 TI - Untargeted LC-Q-TOF mass spectrometry method for the detection of adulterations in skimmed-milk powder. AB - A nontargeted protein identification method was developed to screen for adulterations in skimmed-milk powder (SMP). There are indications of falsified SMP content due to the addition of plant proteins. To demonstrate the reliability and accuracy of the developed comparative LC-MS method using a quadrupole TOF MS instrument, adulterated SMP samples were prepared by the addition of protein isolates of soy and pea to skimmed-milk before pasteurisation and evaporation. The comparative LC-MS approach enabled unequivocal discrimination of those SMP samples containing soy and pea protein from nonadulterated SMP. To identify the source of (plant) proteins present in the adulterated SMP, data-dependent LC MS/MS was used in combination with an include list of differential peptides. Numerous peptides originating from the major seed proteins of soy (glycinin, beta conglycin) and pea (legumin, vicilin) could be identified in this way. PMID- 19301325 TI - On-line multi-enzymatic approach for improved sequence coverage in protein analysis. AB - The development of a new mixed bioreactor for proteomic studies based on trypsin and chymotrypsin is described. Trypsin and chymotrypsin were simultaneously bonded to an epoxy monolithic silica column (100 mmx4.6 mm id) in a one-step reaction via epoxy-groups. In order to compare the catalytic properties of the two enzymes in the isolated and in the multi-enzymatic approach, two other single enzyme bioreactors based on trypsin and chymotrypsin were prepared following the same immobilization protocol. The kinetic parameters of the multi-enzymatic bioreactor were derived and it was demonstrated that it retains the individual catalytic activity of the two enzymes. To prove the power of this experimental approach the new mixed bioreactor was integrated in an LC-ESI-MS/MS system for digestion, enrichment, separation and identification of the test protein insulin like growth factor binding-protein 1 (IGFBP-1). The peptide map and protein sequence coverage obtained with the three bioreactors were compared. The results clearly indicate that the proposed multi-enzyme approach can reduce both digestion and analysis time, accelerate data interpretation and increase the confidence degree in protein identification. PMID- 19301326 TI - Shotgun proteomic analysis of the microsomal fraction of eukaryotic cells using a two-dimensional reversed-phase x ion-pair reversed-phase HPLC setup. AB - A RPxIP-RP HPLC separation scheme was combined with on-line ESI-IT tandem MS or off-line MALDI tandem TOF MS and applied to the analysis of eukaryotic subcellular proteomes. Previous proteomic studies [1] were complemented by the approval of the approach to eukaryotic proteomes using the fission yeast Schizosaccharomyces pombe. The major focus was set to the analysis of primary human hepatocyte microsomes, representing a compartment of high interest due to its involvement in xenobiotic detoxification and cholesterol homeostasis. Of the 588 proteins identified from two donors, 24% are involved in cholesterol homeostasis or xenobiotic/lipid metabolism. Up to 50% of the identified proteins belong to the group of membrane proteins, difficult to investigate using gel based proteomic approaches. We further demonstrated the reproducibility and comparability of the approach and reduced the amount of sample load by almost 70% with only minor loss of information about the proteins identified in the samples. The presented study clearly demonstrates the good applicability of the experimental setup to the analysis of subcellular proteomes including large membrane fractions, where only low amounts of sample material are available. PMID- 19301327 TI - High peak capacity separation of peptides through the serial connection of LC shell-packed columns. AB - High peak capacity was obtained for the separation of a HSA tryptic digest through the serial connection of three LC columns packed with shell material (C18, 15 cmx4.6 mm, 2.7 mum particle size). A major benefit of these particles, consisting of a 1.7 mum solid core and a 0.5 mum porous shell, is the small diffusion path, which reduces axial dispersion of solutes and minimizes peak broadening allowing for higher resolving power to be obtained. Increase in the temperature decreases the viscosity and the backpressure on the columns and in this way allowed to couple multiple columns. Single and multicolumn systems were first evaluated for the separation of a standard sample mixture in the isocratic mode; afterwards three columns could be coupled at 60 degrees C, generating more than 90,000 effective plates (200,000 plates/meter) for the separation of HSA in gradient mode on a conventional HPLC instrument. Peak capacity ratios were found in good accordance with the theoretical gain in resolution, i.e. values of 1.37 and 1.67 were measured, when doubling and tripling the length of stationary phases, respectively (the highest measured peak capacity on three coupled columns was 367). PMID- 19301328 TI - Phosphine-free palladium-catalyzed allene carbopalladation/allylic alkylation domino sequence: a new route to 4-(alpha-styryl) gamma-lactams. AB - Free to decide: Various 4-(alpha-styryl) gamma-lactams are synthesized in 61-88% yield by a phosphine-free palladium-catalyzed carbopalladation/allylic alkylation domino sequence (see scheme). The cyclization is totally regio- and diastereoselective in favor of the 3,4-trans-disubstituted gamma-lactam. The process is successfully applied to the synthesis of a new aza analogue of the naturally occurring lignan (+)-oxo-parabenzlactone. PMID- 19301329 TI - Exploring, tuning, and exploiting the basicity of hydrotalcites for applications in heterogeneous catalysis. AB - Basic catalysis! The basic properties of hydrotalcites (see picture) make them attractive for numerous catalytic applications. Probing the basicity of the catalysts is crucial to understand the base-catalysed processes and to optimise the catalyst preparation. Various parameters can be employed to tune the basic properties of hydrotalcite-based catalysts towards the basicity demanded by each target chemical reaction.Hydrotalcites offer unique basic properties that make them very attractive for catalytic applications. It is of primary interest to make use of accurate tools for probing the basicity of hydrotalcite-based catalysts for the purpose of 1) fundamental understanding of base-catalysed processes with hydrotalcites and 2) optimisation of the catalytic performance achieved in reactions of industrial interest. Techniques based on probe molecules, titration techniques and test reactions along with physicochemical characterisation are overviewed in the first part of this review. The aim is to provide the tools for understanding how series of parameters involved in the preparation of hydrotalcite-based catalytic materials can be employed to control and adapt the basic properties of the catalyst towards the basicity demanded by each target chemical reaction. An overview of recent and significant achievements in that perspective is presented in the second part of the paper. PMID- 19301330 TI - Unprecedented sulfone-functionalized metal-organic frameworks and gas-sorption properties. AB - Gas storage: A new, sulfone-functionalized dicarboxylate-based ligand (see figure) is capable of directing the formation of novel metal-organic frameworks with unprecedented organic and inorganic secondary building units. A high CO(2) uptake with remarkable selectivity over CH(4), N(2), and H(2) was observed at near-ambient temperature. PMID- 19301331 TI - Stereoselective synthesis of polysubstituted alkenes through a phosphine-mediated three-component system of aldehydes, alpha-halo carbonyl compounds, and terminal alkenes. AB - Geometrical control: PPh(3) and methyl acrylate (or acrylamide) are able to mediate the one-pot Wittig reaction of aldehydes with alpha-halo carbonyl compounds for the synthesis of 1,2-disubstituted and trisubstituted alkenes in an excellent stereoselective fashion. Furthermore, the first one-pot, three component reaction of aldehydes, alpha-halo acetates, and terminal alkenes has been developed in the presence of PPh(3) to produce trisubstituted alkenes with excellent E selectivity (see scheme). PMID- 19301332 TI - Rhodium-catalyzed formation of stereocontrolled trisubstituted alkenes from Baylis-Hillman adducts. AB - Efficient and general conditions for the formation of stereodefined trisubstituted alkenes by using the rhodium-catalyzed reaction of unactivated Baylis-Hillman adducts with either organoboronic acids or potassium trifluoro(organo)borates are reported (see scheme).We report here efficient and general conditions for the formation of stereodefined trisubstituted alkenes using the rhodium-catalyzed reaction of unactivated Baylis-Hillman adducts with either organoboronic acids and potassium trifluoro(organo)borates. The use of the [{Rh(cod)OH}(2)] precursor gave very fast coupling reactions under low catalyst loading, very mild reaction conditions (from room temperature up to 50 degrees C) and without the need of additional phosphane ligands. Based on the new reaction conditions, the reaction, originally limited to Baylis-Hillman adducts derived from esters, could be extended to a large variety of Baylis-Hillman adducts, bearing either keto, cyano or amido functionalities. Moreover, the reaction of Baylis-Hillman adducts bearing esters functionality was improved and could be conducted at lower temperature using lower catalyst loading. PMID- 19301333 TI - [2,3]-Wittig rearrangement of enantiomerically enriched 3-substituted 1 propenyloxy-1-phenyl-2-propen-1-yl carbanions: effect of heteroatoms and conjugating groups on planarization of an alpha-oxy-benzylcarbanion through a double bond. AB - Don't get trapped: The effect of conjugating electron-withdrawing groups and alpha-anion-stabilizing heteroatom substituents on configurational stability of chiral carbanions through a double bond was examined on the basis of extent of chirality transfer in intramolecular trapping in [2,3]-Wittig rearrangement of chiral 3-substituted 1-propenyloxy-1-phenyl-2-propen-1-yl carbanions (see scheme).The effect of conjugating electron-withdrawing groups and alpha-anion stabilizing heteroatom substituents on configurational stability of chiral carbanions through a double bond was examined on the basis of extent of chirality transfer in intramolecular trapping in [2,3]-Wittig rearrangement of chiral 3 substituted 1-propenyloxy-1-phenyl-2-propen-1-yl carbanions. PMID- 19301334 TI - Sensing abilities of crown ether functionalized polythiophenes. AB - Tuning sensing abilities! The affinity of three different [15]crown-5 ether functionalized polythiophenes for alkali ions has been explored (see figure). Ab initio and DFT quantum mechanical calculations show that the binding energy between neutral conducting polymers and metallic ions, which interact attractively, decreases as the size of the ion increases. Oxidation of these polythiophene derivatives significantly reduces their affinity towards alkali ions, becoming low or even nonexistent.This work reports a theoretical study of the affinity of three different [15]crown-5 ether functionalized polythiophenes for alkali ions. Ab initio and DFT quantum mechanical calculations have evidenced that the binding energy between neutral conducting polymers and metallic ions, which interact attractively, decreases as the size of the ion increases. However, the preferences of these polythiophene derivatives have been found to be influenced by the entropic contribution, which favors binding with Na(+) in preference to Li(+). Molecular dynamics simulations have shown that this behavior can be accounted for on the basis of the large fluctuations that Li(+) undergoes when it is contained in the cavity of the [15]crown-5 ether macrocycle. In contrast, the fluctuations detected for complexes with Na(+) and K(+) were particularly low, even though only the former ion fits into the center of the macrocyclic cavity. Oxidation of these polythiophene derivatives produces a drastic change in their sensing abilities, their affinities towards alkali ions becoming low or even nonexistent. Moreover, calculations indicate that the presence of unsubstituted thiophene rings directly attached to a functionalized thiophene modulates the strength of the binding in the oxidized state by reducing the repulsive interaction between the metallic cation and the pi-conjugated system. On the other hand, the influence of alkali ions on the electronic properties, in particular, on the ionization potentials and the lowest pi-pi* transitions, of both the neutral and oxidized polythiophenes is discussed. PMID- 19301335 TI - A cobalt metallacrown anion host with guest-dependent redox activity. AB - Two bowl-shaped cavities, each having three OH(-) hydrogen-bond donors at its base, are present in double-cone-shaped metallacrown anion host [Co(6)(mu OH)(6)(mu-L)(6)](m+) (1(m+); HL=3{5}-(pyrid-2-yl)-5{3}-(tert-butyl)pyrazole). Depending on its affinity for the anions present, it can be isolated in its Co(III) (3)Co(II) (3) (m=3; e.g., 1(ClO(4))(3)) and Co(III) (2)Co(II) (4) (m=2; e.g., 1(BF(4))(2)n H(2)O) oxidation states. See picture for photographs of isolated salts.Reaction of cobalt(II) salts with one equivalent of 3{5}-(pyrid-2 yl)-5{3}-(tert-butyl)pyrazole (HL) and NaOH in EtOH affords salts of [Co(6)(mu OH)(6)(mu-L)(6)](m+) (2(m+), m=2 or 3). This is formed from alternating tetrahedral and octahedral cobalt centers, giving a double-cone-shaped molecule with two bowl-shaped cavities. Each cavity has three OH(-) hydrogen-bond donors at its base, giving them an affinity for inorganic anions. Reactions with CoX(2) (X(-)=ClO(4) (-) or CF(3)SO(3) (-)) afford products with the stoichiometry 2X(3). In contrast, when salts of fluorinated anions are used, the products have the formulae 2Y(2)n H(2)O (Y(-)=BF(4) (-), PF(6) (-), or SbF(6) (-); n approximately 2). The connectivity and topology of the metallacrown were confirmed by crystal structures of three of these salts. Magnetic measurements imply that the tricationic products have the oxidation state Co(III) (3)Co(II) (3), while the dications are formulated as Co(III) (2)Co(II) (4). Solutions of 2X(3) contain purely 2(3+) according to NMR spectroscopy, ES mass spectrometry, and UV/Vis/NIR spectroscopy, but solutions of 2Y(2)n H(2)O contain more than one significant component. The 2(3+)/2(2+) couple in MeCN is irreversible at room temperature by cyclic voltammetry, occurring near -1.1 V versus ferrocene/ferrocenium. Importantly, the 2Y(2)n H(2)O compounds do not exhibit this process in the presence of 0.1 M [NBu(4)][BPh(4)] as base electrolyte, but they do show it when the appropriate [NBu(4)]Y salt is used (where the Y(-) ions are present in 10(3) fold excess). Conversely, the 2X(3) complexes exhibit a strong 2(3+)/2(2+) reduction under both experimental conditions. Hence, the isolation of 2(3+) or 2(2+) in the presence of different anions appears to be controlled by the affinity of those anions for the metallacrown. PMID- 19301336 TI - The search for the mechanism of the reaction catalyzed by farnesyltransferase. AB - Atomic-level portrait: The mechanism of the reaction catalyzed by the puzzling enzyme farnesyltransferase is elucidated by using computational methods, allowing the obtainment of the first real detailed atomistic quantum-chemical transition state structure (see figure) for the reaction catalyzed by this enzyme. The results obtained provide an atomic-level framework for the design of more potent and specific inhibitors for this important enzyme. PMID- 19301337 TI - Synthesis, electronic properties, and reactivity of phospholes and 1,1' biphospholes bearing 2- or 3-thienyl C-substituents. AB - Two series of phospholes and 1,1'-biphospholes bearing either 2- or 3-thienyl substituents at the C atoms are prepared by using the Fagan-Nugent route. Their optical (UV/Vis absorption, fluorescence spectra) and electrochemical properties are systematically evaluated. Of particular interest, the first ever reported 3 thienyl-substituted phospholes exhibit higher LUMO levels than their 2-thienyl analogues, and show accordingly different physical properties. This study also reveals that the 1,1'-biphospholes exhibit sigma-pi conjugation. The phosphole and 1,1'-biphosphole derivatives bearing 3-thienyl substituents are characterized by X-ray diffraction study. The structure-property relationship established following the experimental data are fully supported by theoretical studies including time-dependent(TD)-DFT spectra. A photocyclization reaction performed on the thioxo- and oxophospholes having 3-thienyl substituents affords a novel ring-fused phosphole-thiophene derivative, which was characterized by an X-ray diffraction study. The structure and electronic properties of this novel dithienophosphole are discussed based on experimental and theoretical data. PMID- 19301338 TI - Design of a full-profile-matching solution for high-throughput analysis of multiphase samples through powder X-ray diffraction. AB - Few solutions that aim to identify crystalline materials from the analysis of powder X-ray diffraction (XRD) data have been reported to date. A careful inspection of the powder XRD data, and the corresponding highlight of specific failures when it has been used for the determination of the crystallographic phases of zeolites among mixtures, has allowed the creation of the recently proposed strategy: adaptable time warping (ATW). Herein, the design process is thoroughly detailed in a step-by-step manner, which allows a deep understanding of the motivations, improvements, and the resulting remarkable properties of our methodology. Because the use of high-throughput (HT) techniques for the discovery or for increasing the breadth of the synthetic routes of new microporous crystalline structures makes the reliability of search-match methods a critical factor to be assessed, a meticulous evaluation of the reliability and the robustness is provided and supported by both empirical comparisons and mathematical proof. The results offered by our methodology, which clearly outperforms the well-established solutions, open the way towards total automation of such a routine procedure, eliminating laborious and time-consuming controls, preliminary treatments, and settings. Consequently, the proposed solution is of great interest and appears to be very promising, not only because of the numerous potential applications of XRD in materials science, but also the possible expansion of the solution to several other characterization techniques. PMID- 19301339 TI - A first total synthesis of ganglioside HLG-2. AB - A neuritegenic ganglioside from sea cucumber, HLG-2 (see figure), has been synthesized for the first time. The unique tandem of sialic acids, Neu5Gc alpha(2,4)-NeuAc, was established by the combination of a reactive N-Troc sialyl donor and a 1,5-lactamized sialyl acceptor. The ceramide counterpart was assembled in a stereoselective manner. Direct connection of the trisaccharide and the ceramide successfully afforded a precursor of HLG-2, which was converted to ganglioside HLG-2 in pure form.A first synthesis of the neuritegenic ganglioside HLG-2, which was identified in extracts of the sea cucumber Holothuria leucospilota, is described. The characteristic sequence of the trisaccharide part, alpha-N-glycolylsialyl-(2,4)-alpha-N-acetylsialyl-(2,6)-glucoside, was efficiently assembled by coupling of a highly active N-2,2,2 trichloroethoxycarbonyl (Troc)-protected sialyl donor and a 1,5-lactamized sialyl acceptor with high stereoselectivity. The corresponding trisaccharyl imidate donor was directly glycosidated with the primary hydroxyl group of the ceramide part, producing protected HLG-2 in relatively high yield, global deprotection of which furnished ganglioside HLG-2 in highly pure form. PMID- 19301340 TI - Atomically monodispersed and fluorescent sub-nanometer gold clusters created by biomolecule-assisted etching of nanometer-sized gold particles and rods. AB - Atomically monodispersed gold clusters were synthesized by etching gold nanocrystals (particles and rods) with the assistance of biomolecules (amino acids, peptides, proteins, and DNA) under sonication in water. The resulting gold clusters were exclusively composed of eight atoms, as demonstrated by photoluminescence, optical absorption, electrospray ionization mass spectrometry, and transmission electron microscopy measurements. The gold clusters exhibited solvent-dependent photoluminescence properties when exposed to organic solvents, such as chloroform, tetrahydrofuran, or N,N-dimethylformamide, which was attributed to the rich surface properties of the clusters. This strategy, based on top-down etching, offers an approach to create metal clusters from nanomaterials, which show great potential applications in biological labeling/imaging and sensors that utilize photoluminescence properties as the response. PMID- 19301341 TI - Synthesis of alpha-stereogenic amides and ketones by enantioselective conjugate addition of 1,4-dicarbonyl but-2-enes. AB - In the conjugate addition reaction of a alpha,beta-unsaturated compound, the new stereogenic center is created in the beta-position. In contrast, conjugate addition to 1,4-dicarbonyl but-2-enes will generate an alpha-stereogenic center with respect to one of the carbonyl groups, which informally, can be considered as an inversion of normal reactivity patterns or Umpolung. In this paper, we demonstrate that chiral bicyclic guanidine can catalyze the addition of 1,3 dicarbonyl compounds to 1,4-dicarbonyl but-2-enes [(E)-4-oxo-4-arylbutenamides and (E)-4-oxo-4-arylbutenones] with high regioselectivity and enantioselectivity (ee values up to 97%). PMID- 19301342 TI - Closed and open clamlike structures formed by hydrogen-bonded pairs of cyclotricatechylene anions that contain cationic "meat". AB - Clamming up: The hexaphenolic compound cyclotricatechylene, which has a bowl shaped cavity, forms clamlike pairs that encapsulate cations (see picture). Variable hydrogen bonding allows two linked cyclotricatechylene clamshells to be in a closed arrangement when smaller cations such Rb(+) or Cs(+) provide the clam meat, whereas larger cations such as NMe(4) (+) and NEt(4) (+) cause the clam to be partially opened. PMID- 19301343 TI - Ordered poly(p-phenylene)/layered double hydroxide ultrathin films with blue luminescence by layer-by-layer assembly. AB - Lavender layers: A poly(p-phenylene) anionic derivate and exfoliated Mg-Al layered double hydroxide monolayers were assembled into ultrathin films with well defined blue fluorescence (see picture; the numbers indicate the number of bilayers), long-range order, and high photostability. These films work as multiple quantum-well structures for valence electrons. PMID- 19301345 TI - Perfluoropentaphenylborole: a new approach to Lewis acidic, electron-deficient compounds. AB - Zirconocene is the key: A new synthetic method, which utilizes zirconocene mediated coupling of alkynes, has been developed for the preparation of a new class of highly Lewis acidic boroles (see scheme). Such compounds hold potential for applications in catalysis and the field of electron-deficient organic materials. PMID- 19301344 TI - Iterative in situ click chemistry creates antibody-like protein-capture agents. AB - Special agents for protein capture: Iterative in situ click chemistry (see scheme for the tertiary ligand screen) and the one-bead-one-compound method for the creation of a peptide library enable the fragment-based assembly of selective high-affinity protein-capture agents. The resulting ligands are water-soluble and stable chemically, biochemically, and thermally. They can be produced in gram quantities through copper(I)-catalyzed cycloaddition. PMID- 19301346 TI - A microfluidic approach to chemically driven assembly of colloidal particles at gas-liquid interfaces. AB - Bubbling up: Dissolution of CO(2) bubbles in a suspension of colloidal particles chemically induces the assembly of particles on the surface of shrunken bubbles, and thus yields rapid continuous formation of a colloidal armor. This approach maintains the high colloidal stability of particles in bulk, has increased productivity, and allows the formation of bubbles with precisely controlled dimensions. PMID- 19301347 TI - Crystal-structure determination of powdered paramagnetic lanthanide complexes by proton NMR spectroscopy. AB - Shifts for crystals: Solid-state NMR spectroscopy can be used for structure determination of microcrystalline paramagnetic solids at natural isotopic abundance. The protocol makes use of paramagnetic effects, measured on suitably recorded (1)H NMR spectra, to define the conformation of a molecule in the lattice and the intermolecular packing in the solid phase. The method is illustrated with a family of lanthanide compounds (see picture). PMID- 19301348 TI - Valuing health: a new proposal. AB - After criticizing existing systems of health measurement for their unargued commitment to evaluating health states in terms of preferences or well-being, this essay argues that public rather than private values of health states should help guide the allocation of health-related resources. Private evaluation of health states is relative to a prior individual choice of specific activities and goals, while public evaluation is relative to the whole range of important activities and goals. Public evaluation is concerned with securing a wide range of choices as well as with success given one's choice. A reasonable simplification from the public perspective is to focus on just two features of health states: the subjective feelings attached to health states and the limitations that health states imply on the range of important activities that individuals can pursue. Focusing on just these two dimensions permits the construction of a parsimonious classification of health states with regard to what matters most from the public perspective. This classification, which resembles those in the HALex and the Rosser and Kind Disability and Distress Index, might best be built on top of existing health-state classifications, by mapping the health states they define to activity-limitation/feeling pairs. To assign values to these pairs, I propose relying on deliberative groups to make comparisons among the pairs with respect to the relation 'is a more serious limitation on the range of objectives and good lives available to members of the population'. A ranking according to this property, is not a preference ranking, because it is not a ranking in terms of everything that matters to individuals. Working back from the weights attached to the activity-limitation/feeling pairs, one can impute weights for the health states in other classification systems that were mapped to those pairs. If those weights coincide roughly with current weights, then one legitimizes current weights and provides a vehicle for their public discussion and possible revision. If those weights do not coincide, then one has both an argument for revising current views of the cost effectiveness of treatments and policies and a method to carry out such a revision. PMID- 19301349 TI - Prenatal screening and diagnosis of neural tube defects. AB - This review article discusses prenatal screening and diagnosis of neural tube defects (NTD). High detection rates occur in countries operating ultrasound screening programmes because classical two-dimensional ultrasound cranial signs (lemon shaped head, banana cerebellum, ventriculomegaly) are important diagnostic clues to the presence of spina bifida. Careful evaluation of both the spine and a search for other abnormalities is warranted. Important prognostic information for spina bifida relates to the lesion level, with a "watershed" between L3 and L4 marking a very high chance of being wheelchair bound with the higher lesions. Three-dimensional ultrasound using multiplanar views can achieve diagnostic accuracy within one vertebral body in around 80% of patients. There are high rates of pregnancy termination for spina bifida in many European countries, but the use of new imagining techniques allow better prediction of outcome, and consequently a refinement of prenatal counselling. PMID- 19301350 TI - Does social capital determine health? Evidence from eight transition countries. AB - There is growing interest in the role of social relationships in explaining patterns of health. We contribute to this debate by investigating the impact of social capital on self-reported health for eight countries from the Commonwealth of Independent States. We rely on three indicators of social capital at the individual level (trust, participation in local organisations, social isolation) and employ alternative procedures to estimate consistently the impact of social capital on health. The three social capital indicators are choice variables and are hence, by definition, endogenously determined. We attempt to circumvent the endogeneity problems by using instrumental variable estimates. Our results show that the individual degree of trust is positively and significantly correlated with health, this being true with least squares estimators as well as when relying on instrumental variable estimators with (and without) community fixed effects. Similarly, social isolation is negatively and significantly associated with health, irrespective of the procedure of estimation. On the other hand, the effect of being a member of a Putnamesque organisation is more ambiguous and usually not significantly related to health. PMID- 19301351 TI - Magnetic resonance spectroscopy of the fetal brain. AB - Fetal magnetic resonance spectroscopy (MRS) is technically feasible in utero and demonstrates similar findings to those observed in neonatal populations. MRS can provide additional information to conventional T1- and T2-weighted imaging of the fetal brain. It is of particular use when subtle changes are present on conventional fetal MRI sequences, and when imaging fetuses at risk of brain injury and metabolic abnormalities. PMID- 19301352 TI - Double anchoring balloon technique for recanalization of coronary chronic total occlusion by retrograde approach. AB - A retrograde approach through the collateral channels is considered to improve the success rate of percutaneous coronary intervention for coronary chronic total occlusion (CTO). Various kinds of strategies and techniques are required to improve the success rate of this novel approach. We describe a case in which a CTO was successfully recanalized by the retrograde approach, using a new anchoring balloon technique (double anchoring balloon technique) which enabled the successful balloon passage through the hard CTO lesion. PMID- 19301353 TI - Percutaneous closure of a periprosthetic leakage after mitral valve reoperation due to recurrent endocarditis. AB - Paravalvular leakage following the atrioventricular valve replacement, though mostly harmless with insignificant morbidity, can result in heart failure and significant hemolysis that requires treatment. Reoperation is still the treatment of choice, but there is a high risk of recurrence, especially in patients with a history of endocarditis and/or those who have already undergone reoperation for paravalvular leakage. Recently, percutaneous closure of perivalvular leaks with occluders or coils have become an alternative to surgery. However, up to now, the collective of patients who benefit from this approach still has to be defined. Here, we present a case of a highly symptomatic 64-year-old male with severe hemolysis caused by paravalvular leakages after reoperation of a mechanical mitral valve replacement due to recurrent endocarditis. PMID- 19301354 TI - Delayed and repeated cardiac tamponade following microleak in RCA successfully treated with intra arterial sterile glue injection. AB - Postinterventional pericardial leaks mostly occur intraprocedure or immediately postprocedure and call for an aggressive management right then. Also, once controlled for 24-48 hr, the leaks usually seal themselves spontaneously. We herein describe an unusual case of delayed and continued pericardial leak over 10 days as a result of micro perforation of RCA from the distal wire tip and which was successfully managed by intracoronary injection of sterile synthetic glue, which to the best of our knowledge is the first report of its kind. PMID- 19301355 TI - Left coronary artery thrombus characterized by a fully automatic three dimensional gated reconstruction. AB - Rotational coronary angiography and subsequent automatic modeling or reconstruction can result in clinically valuable three-dimensional (3D) representations of the coronaries. From these 3D representations information can be derived for specific coronary segments, such as lesion length, vessel diameter, bifurcation angles, and optimal viewing angles. In this case report, we highlight the characterization of a left coronary artery thrombus by a fully automatic 3D gated reconstruction. This case also shows that detailed 3D morphology of a lesion can be assessed during percutaneous coronary interventions using rotational coronary angiography and subsequent automated image processing. PMID- 19301356 TI - Intravascular ultrasound and 3D angle measurements of coronary bifurcations. AB - OBJECTIVE: To standardize the intravascular ultrasound (IVUS) analysis of coronary bifurcations. BACKGROUND: Percutaneous treatment of bifurcation lesions is difficult particularly at the side branch ostium. Imaging techniques may improve our understanding of treatment options. There is no established IVUS methodology to assess the bifurcation. The present study aims to develop standards for bifurcation imaging. METHODS: Quantitative IVUS analysis and 3D bifurcation angle measurements were performed in 34 patients who were selected from the Washington Hospital Center Database. Patients were included if both left anterior descending (LAD) and first diagonal (DX) pullbacks in the same procedure were done. Angiograms were available in 27 patients to measure the 3D bifurcation angle using specialized software. Pullbacks were analyzed proximal and distal to the bifurcation, and at the bifurcation. RESULTS: Prox(LAD) versus Prox(LAD(DX)) were similar for vessel area (15.5 +/- 4.6 mm(2) vs. 15.9 +/- 4.0 mm(2), P = 0.19), lumen area (8.3 +/- 3.6 mm(2) vs. 8.6 +/- 3.3 mm(2), P = 0.25), and plaque area (7.2 +/- 2.0 mm(2) vs. 7.3 +/- 1.9 mm(2), P = 0.55). However, Bifurcation(LAD) was larger than Bifurcation(DX) for vessel area (17.3 +/- 4.0 mm(2) vs. 16.6 +/- 3.9 mm(2), P = 0.0083). The 3D angiographic bifurcation angle was 50 degrees +/- 13 degrees (range of 26 degrees -84 degrees), and did not affect the IVUS measurements. IVUS analysis showed that bifurcation lesions did obey Murray's Law, as Prox(LAD) lumen area measured 36.7 +/- 25.1 mm(3) versus Dist(LAD)/Dist(DX) measured 38.0 +/- 29.1 mm(3), P = 0.56. CONCLUSIONS: Two IVUS pullbacks should be performed for a complete assessment of the bifurcation and comparison with Murray's Law. The proposed IVUS analysis was not influenced by the bifurcation angle. PMID- 19301357 TI - Statisticians in European regulatory agencies. AB - The UK body of statisticians in the pharmaceutical industry, PSI, has called on heads of European regulatory agencies responsible for assessing applications for marketing authorizations for new medicines in the EU to employ full time statisticians. In order to assess the present situation a survey has been conducted to identify the number of agencies employing one or more full time statisticians. Out of 29 responding agencies, 12 employed one or more statisticians on a full time basis, whereas 17 did not. Among these 17, 7 involved external experts on a regular basis, 5 involved external statisticians on a case-by-case basis, whereas 5 never involved external statistical expertise. Failure to involve statisticians in the assessment of efficacy and safety of medicines does not automatically lead to reports of low quality or invalid assessment of benefit-risk. However, in depth knowledge of statistical methodology is often necessary to uncover weaknesses and potentially biased efficacy estimates. This might be of importance for the final opinion on granting a marketing authorization, and statistical review should therefore be conducted by those who are professionally expert in the area. A positive trend toward an increased involvement of statistical expertise in the European network of regulatory agencies is observed. PMID- 19301358 TI - Effusion cytomorphology and immunocytochemistry of malignant melanoma: five cases of melanotic melanoma and one case of amelanotic melanoma. AB - Effusion cytological analyses of amelanotic malignant melanoma (AMM) are very rare and no concise description of AMM related cytomorphologic features using effusion have been reported. Here, we report the cytomorphological, immunohistochemical, and immunocytochemical findings in the effusion cytology of six cases of malignant melanoma (MM), one case of AMM, and five cases of melanotic malignant melanoma. Papanicolaou-stained smears exhibited conspicuous nucleoli, multinucleation, and cytoplasmic vacuolization in all of the MM cases. In addition, the AMM case displayed numerous mitotic figures and intranuclear cytoplasmic inclusions. With regard to the immunohistochemistry findings, all six cases of melanoma were positive for Melan-A/MART-1, HBME-1, and S-100. In the immunohistochemistry analyses, five of six cases of melanoma were positive for WT 1, as was the AMM specimen. Furthermore, because the effusion analysis of malignant mesothelioma proved positive for WT-1, it should be noted that WT-1 effusion analysis is not an appropriate means to distinguish between MM and malignant mesothelioma. We suggest that it is important to recognize cytomorphologic characteristics, such as melanin pigment, conspicuous nucleoli, multinucleation, and cytoplasmic vacuolization, and to choose appropriate antibodies for the correct diagnosis of MM in effusion. PMID- 19301359 TI - The relative influence of phosphorylation and methylation on responsiveness of peptides to MALDI and ESI mass spectrometry. AB - Qualitative and quantitative analysis of post-translational protein modifications by mass spectrometry is often hampered by changes in the ionization/detection efficiencies caused by amino acid modifications. This paper reports a comprehensive study of the influence of phosphorylation and methylation on the responsiveness of peptides to matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) mass spectrometry. Using well-characterized synthetic peptide mixtures consisting of modified peptides and their unmodified analogs, relative ionization/detection efficiencies of phosphorylated, monomethylated, and dimethylated peptides were determined. Our results clearly confirm that the ion yields are generally lower and the signal intensities are reduced with phosphopeptides than with their nonphosphorylated analogs and that this has to be taken into account in MALDI and ESI mass spectrometry. However, the average reduction of ion yield caused by phosphorylation is more pronounced with MALDI than with ESI. The unpredictable impact of phosphorylation does not depend on the hydrophobicity and net charge of the peptide, indicating that reliable quantification of phosphorylation by mass spectrometry requires the use of internal standards. In contrast to phosphorylation, mono- and dimethylated peptides frequently exhibit increased signal intensities in MALDI mass spectrometry (MALDI-MS). Despite minor matrix-dependent variability, MALDI methods are well suited for the sensitive detection of dimethylated arginine and lysine peptides. Mono- and dimethylation of the arginine guanidino group did not significantly influence the ionization efficiency of peptides in ESI-MS. PMID- 19301360 TI - Isolated pulmonary Langerhans cell histiocytosis with recurrent bilateral pneumothoraces treated with chemotherapy and chemical pleurodesis. PMID- 19301361 TI - From medical image computing to computer-aided intervention: development of a research interface for image-guided navigation. AB - BACKGROUND: This paper describes the development and application of a research interface to integrate research image analysis software with a commercial image guided surgery navigation system. This interface enables bi-directional transfer of data such as images, visualizations and tool positions in real time. METHODS: We describe both the design and the application programming interface of the research interface, as well as showing the function of an example client program. The resulting interface provides a practical and versatile link for bringing image analysis research techniques into the operating room (OR). RESULTS: We present examples from the successful use of this research interface in both phantom experiments and real neurosurgeries. In particular, we demonstrate that the integrated dual-computer system achieves tool-tracking performance that is comparable to the more typical single-computer scenario. CONCLUSIONS: Network interfaces for this type are viable solutions for the integration of commercial image-guided navigation systems and research software. PMID- 19301362 TI - DEA-based efficiency evaluation of a novel robotic system for femoral neck surgery. AB - BACKGROUND: Efficiency evaluation is important for expanding the acceptance and deployment of medical robots, but it is still an open issue. In this study, a data envelopment analysis (DEA)-based method was proposed and implemented with a novel robotic system for femoral neck surgery. METHODS: Femoral neck surgeries with and without the assistance of the robotic system were modelled as decision making units (DMUs). C(2)R model was chosen to calculate the relative efficiency. RESULTS: Clinical simulation experiments were performed with 12 Sawbone models and 24 cadaveric femurs, and 19 clinical cases were chosen to be the control group. The minimum value of the robotically assisted surgery was 1, and the minimum value of the traditional surgery was 0.741535, demonstrating the efficiency of the robotic system. CONCLUSION: This research showed that DEA-based efficiency evaluation is practical for medical robotic systems. PMID- 19301363 TI - Quantitative gray-scale analysis in skeletal muscle ultrasound: a comparison study of two ultrasound devices. AB - Muscle ultrasound is a useful technique to detect neuromuscular disorders. Quantification of muscle echo intensity (EI) using gray-scale analysis is more reliable and more sensitive compared with visual evaluation of the images. We devised a method to reliably use EI normal values established with one ultrasound device for use with another device. Based on measurements in a dedicated phantom and in 7 healthy subjects, a conversion equation was calculated to convert the mean EI. The reliability of this equation was next evaluated in a follow-up study of 22 healthy children. Mean muscle EI could be reliably converted from one ultrasound device to another. This allows for normal values obtained with one device to be used with other devices, which is an important step forward toward the use of quantitative muscle ultrasound in daily clinical care. PMID- 19301364 TI - Behavior of fascicles and the myotendinous junction of human medial gastrocnemius following eccentric strength training. AB - This study is the first in which measurements of thickness, fascicle angle and length, and tendon elongation were combined to examine the impact of eccentric strength training on both muscle architecture and tendinous structures. Eighteen healthy male subjects were divided into an eccentric strength training group (n = 10) and a control group (n = 8). The training program consisted of 18 sessions of eccentric exercises over a 7-week period. All subjects were tested at baseline and after the last training session. Using ultrasound imaging, the fascicle angle and length and thickness of the medial gastrocnemius (MG) were analyzed at rest (i.e., theta(p), Fl(p), and t(p), respectively), at 50% of maximal voluntary contraction (MVC) (i.e., theta(50), Fl(50), and t(50), respectively), and during MVC (i.e., theta(100), Fl(100), and t(100), respectively). Tendon elongation (TE) was measured by tracking the proximal displacement of the myotendinous junction of the MG during ramp isometric contraction. During ramp isometric contraction, the slope of the load-deformation relationship of the gastrocnemius tendon above 50% MVC was defined as an index of stiffness. After training, muscle thickness and fascicle angle increased significantly at rest and during contraction, whereas fascicle length increased at rest and did not change during contraction. Furthermore, the stiffness of the gastrocnemius tendon increased significantly. The results suggest that the behavior of muscle architecture and tendon mechanical properties are affected differently by strength training. PMID- 19301365 TI - Comparison of electromyographic responses for the superficial quadriceps muscles: cycle versus knee-extensor ergometry. AB - The purpose of this study was to compare the electromyographic (EMG) amplitude and mean power frequency (MPF) versus power output relationships for the three superficial quadriceps muscles during incremental cycle (CE) and knee-extensor (KE) ergometry in the same subjects. Eight men performed incremental CE and KE tests to exhaustion. Surface EMG signals were recorded simultaneously from the vastus lateralis (VL), rectus femoris (RF), and vastus medialis (VM). Polynomial regression analyses on a subject-by-subject basis indicated that the relationship between EMG amplitude versus power output was best-fit with either a linear, quadratic, or cubic model for CE, whereas the relationship was best-fit with a linear model for all subjects for the KE test. No consistent relationship was found for EMG MPF within subjects and between muscle groups for CE or KE. Compared with CE, however, the EMG amplitude during KE exercise was, on average, approximately 87% and 30% higher for the RF and VM muscles, respectively. These results suggest that KE exercise may be a better mode of examining EMG amplitude in the quadriceps muscle during incremental exercise than traditional CE. PMID- 19301366 TI - Proteomics of regulated secretory organelles. AB - Regulated secretory organelles are important subcellular structures of living cells that allow the release in the extracellular space of crucial compounds, such as hormones and neurotransmitters. Therefore, the regulation of biogenesis, trafficking, and exocytosis of regulated secretory organelles has been intensively studied during the last 30 years. However, due to the large number of different regulated secretory organelles, only a few of them have been specifically characterized. New insights into regulated secretory organelles open crucial perspectives for a better comprehension of the mechanisms that govern cell secretion. The combination of subcellular fractionation, protein separation, and mass spectrometry is also possible to study regulated secretory organelles at the proteome level. In this review, we present different strategies used to isolate regulated secretory organelles, separate their protein content, and identify the proteins by mass spectrometry. The biological significance of regulated secretory organelles-proteomic analysis is discussed as well. PMID- 19301367 TI - Lung and respiratory muscle function in facioscapulohumeral muscular dystrophy. AB - Pulmonary dysfunction is not a well-recognized feature of facioscapulohumeral muscular dystrophy (FSHD). The aim of this study was to establish the prevalence and type of pulmonary and respiratory muscle dysfunction in FSHD. Sixteen patients with moderately advanced FSHD and 16 healthy controls were evaluated. Standard lung and respiratory muscle function tests were performed. Diaphragm muscle inspiratory action was evaluated with transdiaphragmatic pressure measurements. Lung function tests showed an increased residual volume in five patients. There was a significant difference in global respiratory muscle function in patients versus controls; weakness was mild, and it affected expiratory more than inspiratory muscles. There was no significant difference in the diaphragm inspiratory action of patients versus controls. The dystrophic process that underlies FSHD did not significantly involve the muscles of the diaphragm, but it caused mild global respiratory muscle weakness that affected expiratory more than inspiratory muscles. It is probably not necessary to routinely monitor respiratory muscle function in ambulant FSHD patients who lack symptoms or signs of respiratory impairment. PMID- 19301368 TI - Sarcoplasmic-endoplasmic-reticulum Ca2+-ATPase and calsequestrin are overexpressed in spared intrinsic laryngeal muscles of dystrophin-deficient mdx mice. AB - In the mdx mouse model of Duchenne muscular dystrophy, the lack of dystrophin is associated with increased calcium levels and skeletal muscle myonecrosis. The intrinsic laryngeal muscles (ILM) are protected and do not undergo myonecrosis. We investigated whether this protection is related to an increased expression of calcium-binding proteins, which may protect against the elevated calcium levels seen in dystrophic fibers. The expression of sarcoplasmic-endoplasmic-reticulum Ca(2+)-ATPase and calsequestrin was examined in ILM and in nonspared limb muscles of control and mdx mice using immunofluorescence and immunoblotting. Dystrophic ILM presented a significant increase in the proteins studied when compared to controls. The increase of Ca(2+)-handling proteins in dystrophic ILM may permit better maintenance of calcium homeostasis, with the consequent absence of myonecrosis. The results further support the concept that abnormal Ca(2+) handling is involved in dystrophinopathies. Muscle Nerve, 2009. PMID- 19301369 TI - Dexamethasone-conjugated polyethylenimine as an efficient gene carrier with an anti-apoptotic effect to cardiomyocytes. AB - BACKGROUND: Dexamethasone is a potent glucocorticoid with anti-inflammatory effects. Dexamethasone can protect ischemic cardiomyocytes from apoptosis. To apply the anti-apoptotic effect of dexamethasone to ischemic disease gene therapy, dexamethasone-conjugated polyethylenimine (PEI-Dexa) was synthesized and evaluated as an anti-apoptotic gene carrier. METHODS: PEI-Dexa was synthesized with low molecular weight polyethylenimine (PEI2K, 2 kDa). The transfection efficiency and cytotoxicity of PEI-Dexa were evaluated by luciferase assay and the MTT assay. To evaluate the anti-apoptotic effect, PEI-Dexa/DNA complex was transfected into cells and the cells were treated with H(2)O(2). Cell viability and apoptosis level were measured by the MTT assay and caspase-3 assay, respectively. RESULTS: A transfection assay into H9C2 rat cardiomyocytes showed that PEI-Dexa had the highest transfection efficiency at an 8 : 1 weight ratio (PEI-Dexa/DNA). At this ratio, PEI-Dexa had higher transfection efficiency than high molecular polyethylenimine (PEI25K, 25 kDa) and PEI2K. In addition, the cytotoxicity of PEI-Dexa was lower than that of PEI25K. To evaluate the anti apoptotic effect, PEI-Dexa/pSV-Luc or PEI2K/pSV-Luc was transfected into H9C2 cells and the cells were treated with H(2)O(2). PEI-Dexa was found to reduce caspase-3 activity and increase cell viability compared to PEI2K. Heme oxygenase 1 (HO-1) can protect ischemic cardiomyocytes from apoptosis. Therefore, pSV-HO-1 was cloned and transfected into H9C2 cells using PEI-Dexa. The cells transfected with PEI-Dexa/pSV-HO-1 complex had lower caspase-3 activity and higher viability than the cells transfected with PEI-Dexa/pSV-Luc complex after the H(2)O(2) treatment. CONCLUSIONS: PEI-Dexa is an efficient gene carrier with an anti apoptotic effect and may be useful for anti-apoptotic gene therapy in combination with pSV-HO-1. PMID- 19301370 TI - Pediatric hospitalists: training, current practice, and career goals. AB - OBJECTIVE: To determine the range and frequency of experiences, clinical and nonclinical roles, training, work expectations, and career plans of practicing pediatric hospitalists. DESIGN: Mail survey study of a national sample of 530 pediatric hospitalists of whom 67% (N = 338) were from teaching hospitals, 71% (N = 374) were from children's hospitals, 43% (N = 230) were from freestanding children's hospitals, and 69% (N = 354) were from hospitals with >or=250 beds. RESULTS: The response rate was 84%. The majority (54%; N = 211) had been practicing as hospitalists for at least 3 years. Most reported that the pediatric inpatient unit (94%) and inpatient consultation service (51%) were a part of their regular clinical assignment. Most did not provide service in the normal newborn nursery (58%), subspecialty inpatient service (52%), transports (85%), outpatient clinics (66%), or as part of an emergency response team (53%). Many participated in quality improvement (QI) initiatives (84%) and practice guideline development (81%). CONCLUSIONS: This study provides the most comprehensive information available regarding the clinical and nonclinical roles, training, work expectations, and career plans of pediatric hospitalists. However, the field is currently a moving target; there is significant flux in the hospitalist workforce and variation in the roles of these professionals in their clinical and nonclinical work environment. PMID- 19301371 TI - Interhospital transfer of critically ill and injured children: an evaluation of transfer patterns, resource utilization, and clinical outcomes. AB - OBJECTIVE: To describe patterns of transfer, resource utilization, and clinical outcomes associated with interhospital transfer of critically ill and injured children. DESIGN: Secondary analysis of administrative claims data. PARTICIPANTS: Children 0 to 18 years in the Michigan Medicaid program who underwent interhospital transfer for intensive care from January 1, 2002 to December 31, 2004. The 3 sources of transfer from referring hospitals were: emergency department (ED), ward, or intensive care unit (ICU). MEASUREMENTS: Mortality and duration of hospital stay at the receiving hospitals. RESULTS: Of 1643 interhospital transfer admissions to intensive care at receiving hospitals, 62%, 31%, and 7% were from the ED, ward, and ICU of referring hospitals, respectively. Nineteen percent had comorbid illness, while 11% had organ dysfunction at the referring hospital. After controlling for comorbid illness, patient age, and pretransfer organ dysfunction; compared with ED transfers, mortality in the receiving hospital was higher for ward transfers (odds ratio [OR], 1.76; 95% confidence interval [CI], 1.02-3.03) but not for ICU transfers. Also, compared with ED transfers, hospital stay was longer by 1.5 days for ward transfers and by 13.5 days for ICU transfers. CONCLUSION: In this multiyear, statewide sample, mortality and resource utilization were higher among children who underwent interhospital transfer to intensive care after initial hospitalization, compared with those transferred directly from emergency to intensive care. Decision-making underlying initial triage and subsequent interhospital transfer of critically ill children warrants further study. PMID- 19301372 TI - Symmetrical drug-related intertriginous and flexural exanthema after coronary artery angiography. PMID- 19301373 TI - Preserving female fertility following cancer treatment: current options and future possibilities. AB - Children and women of reproductive age are increasingly surviving cancer diagnoses, and therefore long-term quality-of-life issues are of greater importance at the time of diagnosis. Cancer therapies including radiation and chemotherapy can be detrimental to fertility, and therefore many patients are motivated to preserve fertility prior to cancer treatment. The only highly successful method in preserving fertility to date is embryo cryopreservation, which may not be appropriate for some patients due to age, delay in treatment, cancer type and stage, as well as availability of an acceptable sperm donor. Alternative methods including oocyte cryopreservation and ovarian tissue banking may also preserve fertility while providing additional flexibility to patients. In vitro ovarian follicle maturation following tissue banking is one potential approach that would not require a delay in cancer therapy for ovarian stimulation, would not require an immediate sperm donor, and does not carry the risk of reintroducing malignant cells following tissue transplantation. In vitro follicle culture systems have resulted in successful live births in the mouse. However, many challenges must be addressed in translating the system to the human. This review summarizes current approaches to fertility preservation and discusses recent developments and future challenges in developing a human in vitro follicle culture system. PMID- 19301374 TI - Positively false. PMID- 19301375 TI - Treatment with oseltamivir in children hospitalized with community-acquired, laboratory-confirmed influenza: review of five seasons and evaluation of an electronic reminder. AB - BACKGROUND: When initiated within 48 hours of the onset of symptoms, oseltamivir has been shown to reduce severity and length of influenza illness. Few studies have evaluated the use of oseltamivir in patients hospitalized with influenza. OBJECTIVE: To describe the prescribing practices for oseltamivir in children hospitalized with influenza and to evaluate a mechanism to improve the rate of appropriate prescription. DESIGN, SETTING, PATIENTS: Retrospective cohort study of 929 patients aged 21 years or younger hospitalized with community-acquired laboratory-confirmed influenza (CA-LCI) during 5 consecutive seasons (2000-2005). We examined oseltamivir eligibility, which included patients 1 year of age or older with an influenza test result available within 48 hours of symptom onset. During the 2005-2006 season, an observational trial of an electronic reminder was conducted to improve the frequency of oseltamivir prescription. MEASUREMENTS: Oseltamivir prescription. RESULTS: Of 305 patients (32.8%) eligible for treatment with oseltamivir, 49 (16.1% of those eligible) were prescribed oseltamivir during hospitalization. Prescription rates for indications consistent with the US Food and Drug Administration (FDA) approval ("on label") increased from 0% to 37.2% over 5 seasons (P < 0.0001). Prescriptions outside this recommendation ("off label") also increased over 5 seasons (P < 0.0001). Twenty-nine (5%) of 624 patients were treated with oseltamivir off label; 11 were less than 1 year of age. Initiation of a reminder had no impact on prescription (P > 0.05). CONCLUSIONS: Oseltamivir was used infrequently for children hospitalized with influenza. In addition, use inconsistent with the FDA label of oseltamivir occurs. Mechanisms are needed to improve appropriate prescription of oseltamivir. PMID- 19301376 TI - Legionella pneumonia and use of the Legionella urinary antigen test. PMID- 19301377 TI - Out of Africa. PMID- 19301378 TI - Current treatments of chronic immune-mediated demyelinating polyneuropathies. AB - Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), multifocal motor neuropathy (MMN), and anti-myelin-associated glycoprotein (anti-MAG) neuropathy are three demyelinating acquired neuropathies, with distinct responses to immunotherapy. In placebo-controlled, double-blind, randomized trials, intravenous immunoglobulin (IVIg) has been effective for CIDP and MMN, and plasmapheresis has been effective for CIDP. Corticosteroids have been beneficial in controlled trials for CIDP. Other agents, including cyclophosphamide, rituximab, azathioprine, cyclosporine, interferons, fludarabine, mycophenolate mofetil, and etanercept, have been reported to benefit some patients with inflammatory demyelinating neuropathies in case series and case reports. This review examines the use and toxicity associated with these immunotherapy medications in treating patients with chronic immune-mediated demyelinating neuropathies. Muscle Nerve, 2009. PMID- 19301380 TI - Absence of platelet recovery following Helicobacter pylori eradication in childhood chronic idiopathic thrombocytopenic purpura: a multi-center randomized controlled trial. AB - OBJECTIVE: To investigate the effect of Helicobacter pylori eradication on platelet recovery in childhood chronic idiopathic thrombocytopenic purpura (ITP). PATIENTS AND METHODS: A multi-center randomized controlled trial was conducted. Patients aged 4-18 years, diagnosed with chronic ITP, defined by platelet count below 100 x 10(9)/L lasting more than 6 months without identified causes, were enrolled and underwent (13)C-urea breath test for diagnosis of H. pylori infection. Patients who received prednisolone more than 0.5 mg/kg per day or received other platelet-enhancing therapy were excluded. Patients with H. pylori infection were randomized into two groups: treatment and control groups. Treatment group received a standard protocol for H. pylori eradication and repeated (13)C-UBT at 4-6 weeks to confirm successful therapy while the control group received no specific treatment. Monthly platelet count was monitored for 6 months in both groups. Primary outcome was platelet recovery, defined by platelet count over 100 x 10(9)/L for at least 3 months. RESULTS: Of the 55 ITP children, 16 (29.1%) had H. pylori infection. There were no differences in age, sex, duration of disease, platelet count, and the dose of prednisolone between the treatment group (n = 7) and control group (n = 9). One patient in control group was withdrawn due to massive gastrointestinal bleeding requiring a high dose prednisolone. At 6 months, platelet recovery was demonstrated in one patient in the treatment group as well as one in the control group. CONCLUSION: No beneficial effect of H. pylori eradication on platelet recovery in childhood chronic ITP was identified. PMID- 19301381 TI - Persistent gap of incremental charges for obesity as a secondary diagnosis in common pediatric hospitalizations. AB - OBJECTIVE: To use hospitalization data from 2003 to determine whether prior findings, showing higher charges and longer lengths-of-stay (LOSs) for children with obesity versus those without, were stable over time and whether the magnitude of differences was consistent over a 4-year period. METHODS: Using the 2000 and 2003 Agency for Healthcare Research and Quality (AHRQ) Kids Inpatient Database (KID), we examined discharges for the top 4 nonpregnancy-related principal discharge diagnoses for children aged 2-18 years (asthma, pneumonia, affective disorders, and appendicitis), classified as with or without obesity based on the presence of the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) code 278.0x as a secondary diagnosis. We compared mean charges for hospitalizations with obesity listed as secondary diagnosis versus those without. Results are presented in 2003 dollars. RESULTS: Among children's discharges in 2000 and 2003, 1.1% and 1.6%, respectively, listed obesity as a secondary diagnosis. In 2003, for all 4 diagnoses, adjusted mean hospital charges were statistically significantly higher and adjusted mean LOS was statistically significantly longer for discharges with obesity as a secondary diagnosis versus those without. Additionally, the magnitude of the differences for both charges and LOS was generally somewhat greater in 2003 than in 2000 (asthma 9%, pneumonia 17%, affective disorders 121%, and appendicitis 3%) although it did not achieve statistical significance (P > 0.05). CONCLUSIONS: These findings suggest a widening gap of incremental charges and LOS associated with obesity as a comorbidity. This implies a financial imperative for further research to evaluate factors that contribute to greater resource utilization among obese children. PMID- 19301382 TI - Takotsubo cardiomyopathy presenting with dyspnea. AB - Takotsubo cardiomyopathy (TC), or "broken heart syndrome", is an increasingly recognized condition that mimics acute myocardial infarction with morphologically characteristic left ventricular dysfunction in the absence of coronary artery disease. TC is seen almost exclusively in postmenopausal women following extreme emotional or physiologic stress. Although most patients present with chest pain, limited data suggest that African American patients with TC tend to present with atypical symptoms such as dyspnea or nausea. We present a 57 year-old African American female with TC who presented with severe dyspnea following the shooting death of her son. Added to existing data, our case alerts clinicians to consider Takotsubo syndrome in African American patients with atypical presentation. PMID- 19301383 TI - Characteristics of pediatric hospital medicine fellowships and training programs. AB - OBJECTIVE: To explore the structure, components, and training goals of pediatric hospitalist fellowship programs in North America. DESIGN: We constructed a 17 item structured questionnaire to be administered by phone. Questionnaire items focused on documenting goals, training, requirements, and clinical duties of pediatric hospitalist training programs. From February through June 2007, research staff contacted directors of the programs. Responses were analyzed to determine program characteristics, including goals, formal training requirements, clinical rotations, and participation in hospital administrative activities. RESULTS: All 8 training programs completed the survey. There appear to be 2 distinct tracks for pediatric hospitalist training programs: clinical or academic specialization. Currently there are no standards or requirements for fellowship training from an external accrediting body and the curriculum for these programs is likely driven by service requirements and speculation on the needs of a future generation of pediatric hospitalists. The stated goals of the programs were quite similar. Seven reported that the provision of advanced training in the clinical care of hospitalized patients, quality improvement (QI), and hospital administration are central goals of their training program. Six reported training in the education of medical students and residents to be a primary goal, while 5 indicated training in health services research as a primary goal. CONCLUSIONS: Pediatric hospitalist fellowships are in the very early stages of their development. In time, greater structure across institutions will need to be put in place if they are to succeed in becoming a necessary prerequisite to the practice of hospital medicine. PMID- 19301384 TI - Predictors of smoking cessation and relapse after hospitalization for acute coronary syndrome. AB - BACKGROUND: A hospital admission for a serious cardiac event offers a unique opportunity for smoking cessation. Understanding the factors that predict and enhance cessation among smoking cardiac inpatients is important for hospital physicians and clinical staff. STUDY OBJECTIVE: To determine factors that predict smoking cessation, relapse, or continued smoking among posthospitalized cardiac patients who were smoking at the time of admission. SAMPLE: Patients hospitalized with acute coronary syndrome (ACS) were recruited from 5 hospitals in Michigan to participate in a study assessing hospital quality improvement plus at-home health behavior change counseling. MEASUREMENTS: Patient interview data were collected shortly after discharge and 3 and 8 months later to describe patient demographics, clinical characteristics, tobacco use, and other behaviors. Multinomial logit regression was used to predict smoking cessation, relapse, and continued smoking. RESULTS: Of patients smoking at hospitalization who completed both follow-up interviews, 56.8% (n = 111) were not smoking at 8 months. A significant predictor of successful cessation was higher household income (odds ratio [OR] = 4.72; P = 0.003), while having other smokers in the household decreased the odds of cessation (OR = 0.20; P = 0.001). History of depression increased the odds of relapse (OR = 6.38; P = 0.002) and being a lighter smoker decreased the odds (OR = 0.16; P = 0.026). CONCLUSIONS: Although approximately one-half of the smokers in this study reported successful cessation, interventions are still needed to assist all smokers to successfully quit smoking after an ACS hospitalization. Our data suggest targeting follow-up programs to include other family members and using specialized methods for heavy smokers. PMID- 19301387 TI - Patient attitudes toward fertility preservation. AB - The increased survival rates for pediatric cancer patients and for some malignancies that are common in young adults, such as testicular cancer and Hodgkin disease have led to an increased focus on preserving fertility. Research on the psychosocial aspects of cancer-related infertility is a recent development, but we know that both young men and women value parenthood after cancer. At least 75% of survivors who were childless at diagnosis would like future offspring. For those who do not become parents, long-term distress is common. Younger teens may have difficulty assessing whether parenthood will be important to them in the future, and informed consent protocols need to respect their desires rather than deferring too much to parents. We do not know whether parenting a non-biological child (adopted, conceived through third-party reproduction, or a stepchild) reduces distress as much as being able to have one's own genetic offspring. Survivors often have exaggerated concerns about their children's health risks, but still prefer to have biological children if possible. More research is needed on whether participating in fertility preservation reduces long-term distress about cancer-related fertility. Better evidence-based programs to educate families and reduce decisional conflict are needed. PMID- 19301388 TI - Influence of neuroblastoma stage on serum-based detection of MYCN amplification. AB - BACKGROUND: MYCN oncogene amplification has been defined as the most important prognostic factor for neuroblastoma (NB), the most common solid extracranial neoplasm in children. High copy numbers are strongly associated with rapid tumor progression and poor outcome, independently of tumor stage or patient age, and this has become an important factor in treatment stratification. PROCEDURE: By real-time quantitative PCR analysis, we evaluated the clinical relevance of circulating MYCN DNA of 267 patients with locoregional or metastatic NB in children less than 18 months of age. RESULTS: For patients in this age group with INSS stage 4 or 4S NB and stage 3 patients, serum-based determination of MYCN DNA sequences had good sensitivity (85%, 83%, and 75% respectively) and high specificity (100%) when compared to direct tumor gene determination. In contrast, the approach showed low sensitivity patients with stages 1 and 2 disease. CONCLUSION: Our results show that the sensitivity of the serum-based MYCN DNA sequence determination depends on the stage of the disease. However, this simple, reproducible assay may represent a reasonably sensitive and very specific tool to assess tumor MYCN status in cases with stage 3 and metastatic disease for whom a wait and see strategy is often recommended. PMID- 19301389 TI - Double bromodomain-containing gene Brd2 is essential for embryonic development in mouse. AB - The BET subfamily of bromodomain-containing genes is characterized by the presence of two bromodomains and a unique ET domain at their carboxyl termini. Here, we show that the founding member of this subfamily, Brd2, is an essential gene by generating a mutant mouse line lacking Brd2 function. Homozygous Brd2 mutants are embryonic lethal, with most Brd2(-/-) embryos dying by embryonic day 11.5. Before death, the homozygous embryos were notably smaller and exhibited abnormalities in the neural tube where the gene is highly expressed. Brd2 deficient embryonic fibroblast cells were observed to proliferate more slowly than controls. Experiments to explore whether placental insufficiency could be a cause of the embryonic lethality showed that injecting diploid mutant embryonic stem cells into tetraploid wild-type blastocysts did not rescue the lethality; that is Brd2-deficient embryos could not be rescued by wild-type extraembryonic tissues. Furthermore, there were enhanced levels of cell death in Brd2-deficient embryos. PMID- 19301390 TI - Cloning and spatiotemporal expression of zebrafish neuronal nicotinic acetylcholine receptor alpha 6 and alpha 4 subunit RNAs. AB - Acetylcholine plays an important role in regulation of nervous system development and function. We are developing zebrafish (Danio rerio) as a model system to study the role of specific neuronal nicotinic acetylcholine receptor (nAChR) subtypes in development and the effects of nicotine on the developing vertebrate nervous system. We previously characterized the expression of several zebrafish nAChR subunits. To further develop the zebrafish model, here we report a study on the molecular characterization of two additional nAChR subunit genes, designated chrna6 and chrna4. Both zebrafish nAChRs have a high degree of sequence identity to nAChRs expressed in a variety of mammalian species. Reverse transcription polymerase chain reaction was used to show that both nAChR subunit RNAs were expressed early in zebrafish development, with the chrna4 transcript present at 3 hours postfertilization (hpf) and the chrna6 RNA present at 10 hpf. In situ hybridization was used to localize chrna6 and chrna4 RNA expression in 24, 48, 72, and 96 hpf zebrafish. The chrna6 and chrna4 RNAs were each expressed in a unique pattern, which changed during development. At various ages, chrna6 was expressed in Rohon-Beard sensory neurons, trigeminal ganglion, retina, and the pineal gland. Most notably, chrna6 was expressed in catecholaminergic neurons in the midbrain, but was also present in noncatecholaminergic cells in both midbrain and hindbrain. The expression of chrna6 RNA in catecholaminergic cells supports the use of zebrafish as a valid model system to better understand the molecular basis of cholinergic regulation of dopaminergic signaling and the role of alpha6 containing nAChRs in Parkinson's disease. The most notable chrna4 expression was in neural crest cells at 24 hpf and reticulospinal neurons in hindbrain at 48 hpf. chrna4 RNA exhibited a widespread and robust expression pattern in the midbrain in 72 hpf and 96 hpf zebrafish. PMID- 19301391 TI - Dynamic organization and plasticity of sponge bodies. AB - Sponge bodies, cytoplasmic structures containing post-transcriptional regulatory factors, are distributed throughout the nurse cells and oocytes of the Drosophila ovary and share components with P bodies of yeast and mammalian cells. We show that sponge body composition differs between nurse cells and the oocyte, and that the sponge bodies change composition rapidly after entry into the oocyte. We identify conditions that affect sponge body organization. At one extreme, components are distributed relatively uniformly or in small dispersed bodies. At the other extreme, components are present in large reticulated bodies. Both types of sponge bodies allow normal development, but show substantial differences in distribution of Staufen protein and oskar mRNA, whose localization within the oocyte is essential for axial patterning. Based on these and other results we propose a model for the relationship between P bodies and the various cytoplasmic bodies containing P body proteins in the Drosophila ovary. PMID- 19301392 TI - Transcriptional control of Rohon-Beard sensory neuron development at the neural plate border. AB - Rohon-Beard (RB) mechanosensory neurons are among the first sensory neurons to develop, and the process by which they adopt their fate is not completely understood. RBs form at the neural plate border (NPB), the junction between neural and epidermal ectoderm, and require the transcription factor prdm1a. Here, we show that prior to RB differentiation, prdm1a overlaps extensively with the epidermal marker dlx3b but shows little overlap with the neuroectodermal markers sox3 and sox19a. Birthdating analysis reveals that the majority of RBs are born during gastrulation in zebrafish, suggesting that it is during this period that RBs become specified. Expression analysis in prdm1a and neurogenin1 mutant and dlx3b/dlx4b morpholino-injected embryos suggests that prdm1a is upstream of dlx3b, dlx4b, and neurogenin1 at the NPB. mRNA for neurogenin1 or dlx3b/dlx4b can rescue the lack of RBs in prdm1a mutants. Based on these data, we suggest a preliminary gene regulatory network for RB development. PMID- 19301393 TI - The characterization of a zebrafish mid-hindbrain mutant, mid-hindbrain gone (mgo). AB - The vertebrate mid-hindbrain boundary (MHB) is a crucial morphological structure required for patterning and neural differentiation of the midbrain and anterior hindbrain. We isolated a novel zebrafish mutant, MHB gone (mgo), that exhibited a defective MHB. Expression of engrailed3 in the prospective MHB was absent at the 1-somite stage, suggesting that initiation of the isthmic organizer was disrupted in mgo mutants. Complementation test with mgo and noi, in which the pax2a gene is mutated, infer that the mgo mutant may represent a novel noi allele. However, pronephric, otic vesicle, and commissural axonal defects described in noi mutants were not associated with mgo mutants. Genetic mapping revealed that the mgo mutation is linked to the Pax2a locus, but no mutation was detected in pax2a exons or within intron-exon boundaries. Based on these findings, we propose that the mgo mutation genetically interacts with pax2a required for the initiation of MHB formation. PMID- 19301394 TI - Impaired lactation in mice expressing dominant-negative FADD in mammary epithelium. AB - The Fas-associated death domain (FADD/Mort1) adaptor protein was originally identified as a key mediator of apoptosis, although pleiotropic functions for FADD have also been reported. FADD-mediated tumoricidal effects have been described in breast cancer cells; however, its physiological role in normal mammary gland epithelium is not well understood. To determine the role of FADD signaling during mammary gland development, we generated transgenic mice overexpressing dominant-negative FADD (DN-FADD) in mammary epithelium, using the steroid responsive mouse mammary tumor virus promoter. Transgenic mice exhibited a perturbation in lactation resulting in impaired milk production and pup growth retardation. Reduced expansion of alveoli was evident during early lactation with extensive shedding of luminal alveolar cells. Significantly more TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labeling) positive cells were present at this time point and a subsequent increase in bromodeoxyuridine-positive cells was observed. These findings suggest a role for FADD in maintaining the survival of mammary secretory alveolar cells after the establishment of lactation. PMID- 19301395 TI - In vitro embryo culture in defined, sub-microliter volumes. AB - The high attrition rate of in vitro human embryo culture presents a major obstacle in the treatment of clinical infertility by in vitro fertilization (IVF). Physical and genetic requirements are not well understood for human or mouse preimplantation embryo development. Group culture is an established requirement for optimal embryo development in the mouse model. However, conventional microdrop culture limitations hinder investigations of the effects of physical parameters on in vitro embryo development. We report a microfluidics platform that enables embryo culture in precisely defined, sub-microliter volumes (5-500 nl) which cannot be investigated using conventional methods. Groups of two embryos per microfluidic well successfully developed to the blastocyst stage, at a rate of over 80%, which is comparable to those cultured in 20-microl microdrops. This system can be used to dissect physical requirements of in vitro single or group embryo culture, and be made highly parallel to increase experimental throughput. PMID- 19301397 TI - Atypical hemolytic uremic syndrome responsive to steroids and intravenous immune globulin. AB - Atypical hemolytic uremic syndrome remains a challenge to diagnose and treat, with significant acute morbidity and risk for progression to end stage renal disease. Treatment strategies center on plasma exchange but do not necessarily affect the progression of disease. We report the case of a patient with atypical HUS resulting from a mutation in the complement pathway who responded to treatment with steroids and IVIG, therefore avoiding transfusion or plasma exchange. PMID- 19301396 TI - Ex vivo magnetofection: a novel strategy for the study of gene function in mouse organogenesis. AB - Gene function during mouse development is often studied through the production and analysis of transgenic and knockout models. However, these techniques are time- and resource-consuming, and require specialized equipment and expertise. We have established a new protocol for functional studies that combines organ culture of explanted fetal tissues with microinjection and magnetically induced transfection ("magnetofection") of gene expression constructs. As proof-of principle, we magnetofected cDNA constructs into genital ridge tissue as a means of gain-of-function analysis, and shRNA constructs for loss-of-function analysis. Ectopic expression of Sry induced female-to-male sex-reversal, whereas knockdown of Sox9 expression caused male-to-female sex-reversal, consistent with the known functions of these genes. Furthermore, ectopic expression of Tmem184a, a gene of unknown function, in female genital ridges, resulted in failure of gonocytes to enter meiosis. This technique will likely be applicable to the study of gene function in a broader range of developing organs and tissues. PMID- 19301398 TI - Novel markers of early ovarian pre-granulosa cells are expressed in an Sry-like pattern. AB - Mammalian gonad differentiation involves sexually dimorphic cell-fate decisions within the bipotential gonadal primordia. Testis differentiation is initiated by a center-to-poles wave of Sry expression that induces supporting cell precursors (SCPs) to become Sertoli rather than granulosa cells. The initiation of ovary differentiation is less well understood. We identified two novel SCP markers, 1700106J16Rik and Sprr2d, whose expression is ovary-biased during early gonad development, and altered in Wnt4, Sf1, Wt1, and Fog2 mutant gonads. In XX and XY gonads, both genes were up-regulated at approximately E11 in a center-to-poles wave, and then rapidly down-regulated in XY gonads in a center-to-poles wave, which is reminiscent of Sry expression in XY gonads. Our data suggest that 1700106J16Rik and Sprr2d may have important roles in early gonad development, and are consistent with the hypothesis that ovarian SCP differentiation occurs in a center-to-poles wave with similar timing to that of testicular SCP differentiation. PMID- 19301399 TI - A distinct profile of myogenic regulatory factor detection within Pax7+ cells at S phase supports a unique role of Myf5 during posthatch chicken myogenesis. AB - Satellite cells are skeletal muscle stem cells that provide myogenic progeny for myofiber growth and repair. Temporal expression of muscle regulatory factors (MRFs) and the paired box transcription factor Pax7 defines characteristic phases of proliferation (Pax7(+)/MyoD(+)/myogenin(-)) and differentiation (Pax7( )/MyoD(+)/myogenin(+)) during myogenesis of satellite cells. Here, using bromodeoxyuridine (BrdU) labeling and triple immunodetection, we analyzed expression patterns of Pax7 and the MRFs MyoD, Myf5, or myogenin within S phase myoblasts prepared from posthatch chicken muscle. Essentially, all BrdU incorporation was restricted to Pax7(+) cells, of which the majority also expressed MyoD. The presence of a minor BrdU(+)/Pax7(+)/myogenin(+) population in proliferation stage cultures suggests that myogenin up-regulation is alone insufficient for terminal differentiation. Myf5 was detected strictly within Pax7(+) cells and decreased during S phase while MyoD presence persisted in cycling cells. This study provides novel data in support of a unique role for Myf5 during posthatch myogenesis. PMID- 19301400 TI - Kit and foxd3 genetically interact to regulate melanophore survival in zebrafish. AB - We have investigated the role of foxd3 activity in conjunction with signaling by the kit tyrosine kinase receptor in zebrafish black pigment cell (melanophore) development. As loss-of-function of these molecules individually has distinct effects on melanophore number, we have examined the phenotype of double mutants. Individuals with a null mutation in kit have fewer melanophores than wild-type, with cells lost through death. When kit mutants are injected with foxd3 antisense morpholino oligonucleotides or crossed with a foxd3 zebrafish mutant, they have more melanophores than their uninjected or foxd3+ counterparts. Examination of foxd3 loss-of-function in two additional kit mutants that differentially alter kit-dependent migration and survival indicates a change in melanophore number in survival mutants only. Consistently, TUNEL (terminal deoxynucleotidyl transferase mediated deoxyuridinetriphosphate nick end-labeling) analysis confirms a partial rescue of melanophores from cell death. Ectopic expression of foxd3 indicates that foxd3 promotes early melanophore death only when kit is inactive. Taken together, these data suggest a kit-dependent role for foxd3 in the regulation of melanophore survival. PMID- 19301402 TI - Deletion of Pax7 changes the tunica muscularis of the mouse esophagus from an entirely striated into a mixed phenotype. AB - The mechanisms responsible for the different amounts of striated muscle in mammalian esophagi are still enigmatic. A recent ultrastructural analysis in mouse esophagus pointed to a particular role of satellite cells during postnatal growth of striated muscle. The aim of this study was to investigate satellite cell development and the influence of Pax7 on this process. Developing and adult esophagi of wild-type and mice carrying a targeted mutation in Pax7 were analyzed by electron microscopy. We found a gene dose-dependent delayed development of striated muscle and a severe loss of satellite cells in Pax7(+/-) and Pax7(-/-) esophagi. In contrast to the entirely striated wild-type esophagus, Pax7(-/-) mutants developed a mixed phenotype with predominantly smooth muscle caudally. We conclude that Pax7-dependent myogenic progenitor cells are of prime importance for striated muscle formation and the degree of smooth-to-striated muscle conversion during esophageal ontogeny. PMID- 19301403 TI - Defective development of the gall bladder and cystic duct in Lgr4- hypomorphic mice. AB - Leucine-rich repeat (LRR) -containing G protein coupled receptor (LGR) family members are characterized by the presence of a seven-transmembrane domain and LRR motifs. We describe a new function for Lgr4 in the development of the gall bladder and cystic duct and in the epithelium-mesenchyme interaction. Lgr4 expression was observed in the gall bladder epithelium when the gall bladder primordium elongated ventrally. Although Lgr4 hypomorphic mutant (Lgr4(Gt/Gt)) embryos developed a normal gall bladder bud at embryonic day (E) 10.25, no further elongation was observed at later stages. At E12.5, the mesenchyme surrounding the gall bladder had completely disappeared in Lgr4(Gt/Gt) embryos, while the gall bladder remained unelongated. Neighboring tissues such as liver and pancreas were unaffected, as revealed by expression of marker genes. This is the first report of a mutant mouse that lacks a gall bladder and cystic duct without affecting the other tissues that derive from the same hepatic diverticulum. PMID- 19301404 TI - Overlapping functions of Cdx1, Cdx2, and Cdx4 in the development of the amphibian Xenopus tropicalis. AB - Using Xenopus tropicalis, we present the first analysis of the developmental effects that result from knocking down the function of the three Cdx genes present in the typical vertebrate genome. Knockdowns of individual Cdx genes lead to a similar range of posterior defects; compound Cdx knockdowns result in increasingly severe posterior truncations, accompanied by posterior shifts and reduction of 5' Hox gene expression. We provide evidence that Cdx and Wnt3A genes are components of a positive feedback loop operating in the posterior axis. We show that Cdx function is required during later, but not early stages of development, for correct regional specification of the endoderm and morphogenesis of the gut. Our results support the hypothesis that during amphibian development the overall landscape of Cdx activity in the embryo is more important than the specific function of individual Cdx proteins. PMID- 19301405 TI - Toto, I don't think we're on the adult inpatient unit anymore. PMID- 19301406 TI - Fertility preservation medicine: a new field in the care of young cancer survivors. AB - Treatment modalities for numerous oncological and non-oncological conditions result in gonadal insufficiency and infertility. Furthermore, pelvic-abdominal radiation may result in uterine damage resulting in poor reproductive outcomes such as preterm birth, low birth weight, and spontaneous abortion in adult survivors of childhood cancers. In response to the recognition of the impact of cancer treatments on fertility, several fertility preservation techniques have been developed. In prepubertal children, fertility preservation options are usually limited to ovarian cryopreservation because of sexual immaturity, but oocyte freezing can be performed in adolescent children. Two prospective randomized studies showed no benefit of gonadal suppression with GnRH analogs to preserve gonadal function and thus this treatment should not be recommended. For adult survivors of childhood cancer who experienced reproductive failure, third party reproduction techniques are highly successful. PMID- 19301408 TI - Nocturia is an age-independent risk factor for hip-fractures in men. AB - AIMS: Nocturia is a highly prevalent symptom in the elderly and a common reason for interrupted sleep resulting in dizziness, worse daytime functioning and higher risk of falls. The aim of this study was to determine the role of nocturia as a risk factor for hip-fractures in men. METHODS: Men aged 40-80 years participating in a health-screening survey in Vienna between 2000 and 2003 entered the study. In parallel to the investigation all men completed the International Prostate Symptom Score (IPSS). In 2008, files of all Austrian public hospitals were screened whether these men were admitted with the diagnosis of a hip-fracture. Chi(2)-test and logistic regression analyses were used to study the association of nocturia to hip-fractures. RESULTS: A total of 1,820 men (52 +/- 9 years) with a mean follow-up of 6.2 years entered this analysis. Hip fractures occurred in 24 men (1.3%). The occurrence of hip-fractures increased from 0.9% (no nocturia) to 1.0% (nocturia once) to 2.7% (nocturia twice or more). This trend was significant (P = 0.03, chi(2)-test). Even after adjusting for age, men with nocturia of >or=2 were at increased risk (OR 1.36; 95% CI 1.03-1.80, P = 0.03) for developing a hip-fracture. The IPSS was not correlated to the occurrence of hip-fractures (p for trend 0.61). CONCLUSIONS: Nocturia of >or=2 is an age-independent risk factor for hip-fractures in men. The high frequency of nocturia in elderly men paralleled with the significant morbidity and even mortality of hip-fractures underline the clinical importance of this association. PMID- 19301409 TI - Diagnostic delay and disease stage in head and neck cancer: a systematic review. AB - OBJECTIVES/HYPOTHESIS: The objective of this study was to examine the evidence for an association between patient and/or provider-related diagnostic delay and late stage at diagnosis. STUDY DESIGN: We identified all English language published studies worldwide and present a summary of the direction and magnitudes of the associations observed. We consider the role of study population characteristics and symptom variation across the head and neck cancer sites on the delay-stage association. RESULTS: The 27 eligible studies reviewed varied considerably in the cancer types grouped by analysis, types of delay, and measurement of delay. The relationship between diagnostic delay and stage at diagnosis varied in direction and magnitude, with no consistent positive association in any of the head and neck cancer sites. CONCLUSIONS: Possible explanations for the lack of an observed relationship between patient delay and stage include: inaccurate measurement of delay, lack of sensitivity of disease stage to delay-related disease progression, and variation in tumor aggressiveness, which could lead to variation in symptom progression rates. We call for better evidence about the relationship between diagnostic delay and disease progression and/or disease outcomes. If demonstrated and validated, such associations would provide a much stronger argument than description of delay alone for education programs around symptom recognition and for more active screening of high-risk individuals. PMID- 19301410 TI - "We have to stop again?!": The impact of overactive bladder on family members. AB - AIMS: Overactive bladder (OAB), characterized by urinary urgency, with or without incontinence, and urinary frequency and nocturia, has a significant impact on patients' lives. Given the social context inherent in OAB, the purpose of this study was to identify issues faced by OAB family members. METHODS: Incontinent OAB patients and their family members were recruited from a urology clinic. Continent OAB family members were recruited via newspaper advertisements. Focus groups were conducted to evaluate family impact; content analysis was used to analyze data. RESULTS: Participants included 14 dyads consisting of incontinent OAB patients and their family members (partners or daughters) and 17 continent OAB family members. Urinary frequency was the most bothersome symptom for family members as family members reported that their partners' urinary frequency significantly altered their lives by limiting activities (e.g., travel, social activities) and impacting time because of persistent, and urgent, need to find a toilet. Nocturia caused sleep disruption and fatigue for both patients and their partners. Family members indicated that their partner's OAB fostered significant emotions including embarrassment, anxiety, anger, worry, frustration, and sympathy. Several spouses reported that OAB limited their sexual interactions and intimacy. CONCLUSIONS: OAB has a significant impact on OAB patients' family members, regardless of whether the patient and family member live together. Consequently, it is important to assess the family impact of symptomatic conditions such as OAB to fully understand the outcomes of disease and treatment. PMID- 19301411 TI - Cochlear implant surgery in patients more than seventy-nine years old. AB - OBJECTIVES/HYPOTHESIS: To evaluate the surgical complications, auditory performance, and hearing handicap following cochlear implantation in patients greater than 79 years of age. STUDY DESIGN: Retrospective trial, tertiary referral center. METHODS: The study group was comprised of 21 patients implanted after 79 years of age from 1996 through 2006 with follow-ups past their 8th decade. Pre-op evaluation consisted of pure-tone audiometry and speech discrimination scores (Hearing in Noise Test and City University of New York sentence test). The results of these tests were compared to similar tests taken post-op. A validated hearing handicap questionnaire was used to evaluate the outcome. RESULTS: There were no permanent medical or surgical complications. However, two patients developed exacerbations of previous comorbid conditions (i.e., urinary retention and acute delirium). Implanted patients experienced a significant improvement in audiologic performance, post-op pure tone average, and post-op speech scores (P < .001). A majority of them were able to use the phone and reported that the cochlear implant was of great benefit to them. The post-op hearing handicap inventory for the elderly demonstrated a significant decrease of hearing handicap scores. CONCLUSIONS: This is the first study to focus on a patient group this advanced in age. With increasing life expectancy, we should begin to stratify risk versus benefit of cochlear implantation in this age group. Cochlear implantation improved audiologic performance and the quality of life in patients older than 79 years old. There were no permanent medical or surgical complications. Chronic pain and temporary vertigo were the most common complications reported in this elderly group. Laryngoscope, 2009. PMID- 19301412 TI - Dehalogenation of gaseous 1-chlorobutane by dehydrated whole cells: Influence of the microenvironment of the halidohydrolase on the stability of the biocatalyst. AB - It was observed that a biocatalyst prepared from dehydrated whole cells of a recombinant Escherichia coli (initially suspended in borate buffer) was able to hydrolyze gaseous 1-chlorobutane in a solid/gas reactor. Nevertheless, at 40 degrees C and for a 0.7 water activity, it rapidly lost its activity. The explanation of this phenomenon was first investigated by observing the biocatalyst structure at the microscopic level and by studying the localization of the dehalogenase involved in catalysis (intracellular/extracellular). The behavior of this biocatalyst was then compared with that of a preparation made from cells extracts. The reasons of the inactivation are discussed in terms of thermal denaturation and protective effect of buffer salts. PMID- 19301413 TI - Bladder outlet obstruction in painful bladder syndrome/interstitial cystitis. AB - AIMS: Obstructive symptoms such as slow stream, dribbling and straining are often reported by painful bladder syndrome and interstitial cystitis (PBS/IC) patients. Our hypothesis was that some patients with PBS/IC have an associated measurable bladder outlet obstruction (BOO) secondary to dysfunctional voiding and that those patients with more severe PBS/IC are more likely to have BOO. METHODS: This is a retrospective chart review of female patients diagnosed with PBS/IC based on the NIDDK research definition. Charts were reviewed for clinical symptom severity, ulcer or non-ulcer PBS/IC on cystoscopy, and pressure-flow urodynamics (UDPF). Patients were excluded if they had a urinary infection at the time of urodynamics or did not meet study entry requirements. The cut-off values of or=25 cm of water was used to define BOO. RESULTS: Of the 231 women: 38 had ulcer PBS/IC and 193 had non-ulcer PBS/IC. MCC was 269 ml in non ulcer PBS/IC and 200 ml in ulcer PBS/IC (P = 0.006). One hundred eleven women (48%) met criteria for obstruction. MCC was 298 ml in the non-obstructed group and 214 ml in the obstructed group (P < 0.0001). The maximum flow with non-ulcer PBS/IC was 11.0 ml/sec and in ulcer PBS/IC 8.9 ml/sec (P = 0.04) Detrusor pressure at maximum flow was 33.3 cm H(2)O, in non-ulcer, and 37.4 cm H(2)O in ulcer PBS/IC (P = 0.01). CONCLUSIONS: Forty-eight percent of our PBS/IC patients have BOO, and increasing severity of PBS/IC is associated with higher voiding pressure. PMID- 19301414 TI - Pharyngeal morphology: a determinant of successful nasal surgery for sleep apnea. AB - OBJECTIVES/HYPOTHESIS: To estimate the effectiveness of nasal surgery on the occurrence of sleep apnea, and to analyze the pharyngeal morphology of apnea patients whose sleep-disordered breathing was ameliorated postoperatively. STUDY DESIGN: Prospective study. METHODS: Thirty-five consecutive patients with apnea and nasal obstruction underwent polysomnography and a morphological examination of the upper airway before and after nasal surgery, which included septoplasty, inferior turbinectomy, and/or functional endoscopic sinus surgery. RESULTS: Sleep apnea was significantly ameliorated in only eight patients. The postoperative reduction in the apnea-hypopnea index tended to be lower in those with a low positioned soft palate, reflected in an elevated modified Mallampati score, and a narrow retroglossal space. Neither swollen tonsils nor narrow fauces affected the surgical outcome. Regression analysis showed that the modified Mallampati score (P < .05) and the retroglossal space (P < .05) were significant predictors of postoperative improvement in the apnea-hypopnea index. CONCLUSIONS: Among sleep apnea patients suffering from nasal obstruction, nasal surgery is effective in those with a high-positioned soft palate and/or a wide retroglossal space. PMID- 19301415 TI - The effects of sound conditioning on gentamicin-induced vestibulocochlear toxicity in gerbils. AB - OBJECTIVES/HYPOTHESIS: Recent studies in animal models have shown via physiologic and histologic measures that the administration of exogenous antioxidants is protective against gentamicin-induced oto-vestibulo toxicity. In addition, studies have also shown that sound conditioning increases cochlear antioxidants. The objective of this study is to determine whether sound conditioning provides protection against gentamicin in the cochlear and/or vestibular system. STUDY DESIGN: : Prospective animal study. METHODS: Three-month-old gerbils were divided into three groups (A, B, and C). The gerbils in group A were sound conditioned only (n = 2). In group B, the animals received gentamicin on the round window (n = 2). The gerbils in group C were sound conditioned first and later received gentamicin to the round window (n = 2). The animals were ultimately sacrificed and their right cochlea and posterior crista ampullaris were removed, processed, and sectioned. The specimens were analyzed for inner hair cell (IHC) and outer hair cell (OHC) loss and vestibular supporting and sensory hair cell nuclei per micrometer of vestibular epithelium. RESULTS: The sound-conditioned group (A) had no loss of cochlear hair cells. The gerbils treated with gentamicin only (B) had a 34% decrease of OHCs and 49% decrease of IHCs. The sound-conditioned plus gentamicin-treated group (C) had a 5.5% decrease in OHCs and 12% decrease in IHCs. There were no significant differences with regards to supporting cell nuclei within the posterior crista across all groups. When compared to group A, the gerbils in groups B and C did have a 23 to 42% decrease in the number of sensory cell nuclei per micrometer of vestibular epithelium. CONCLUSIONS: Sound conditioning does appear to attenuate the effects of gentamicin in the cochlea, although not significantly altering its vestibulotoxicity. An upregulation of cochlear-specific antioxidants is believed to be an important factor. As we had a small sample size, we can only note trends in the data, but future studies with more animals and measurements of antioxidant levels after sound conditioning would be useful to quantify this effect and determine if it can be exploited clinically. Laryngoscope, 2009. PMID- 19301416 TI - Positron emission tomography imaging of D(2/3) agonist binding in healthy human subjects with the radiotracer [(11)C]-N-propyl-norapomorphine: preliminary evaluation and reproducibility studies. AB - OBJECTIVE: (-)-N-[(11)C]-propyl-norapomorphine (NPA) is a full dopamine D(2/3) receptor agonist radiotracer suitable for imaging D(2/3) receptors configured in a state of high affinity for agonists using positron emission tomography. The aim of the present study was to define the optimal analytic method to derive accurate and reliable D(2/3) receptor parameters with [(11)C]NPA. METHODS: Six healthy subjects (four females/two males) underwent two [(11)C]NPA scans in the same day. D(2/3) receptor-binding parameters were estimated using kinetic analysis (using one- and two-tissue compartment models) as well as simplified reference tissue method in the three functional subdivisions of the striatum (associative striatum, limbic striatum, and sensorimotor striatum). The test-retest variability and intraclass correlation coefficient were assessed for distribution volume (V(T)), binding potential relative to plasma concentration (BP(P)), and binding potential relative to nondisplaceable uptake (BP(ND)). RESULTS: A two tissue compartment kinetic model adequately described the functional subdivisions of the striatum as well as cerebellum time-activity data. The reproducibility of V(T) was excellent (0.75) in the three functional subdivisions of the striatum. Although SRTM led to an underestimation of BP(ND) values relative to that estimated by kinetic analysis by 8-13%, the values derived using both the methods were reasonably well correlated (r(2) = 0.89, n = 84). Both methods were similarly effective in detecting the differences in [(11)C]NPA BP(ND) between subjects. CONCLUSION: The results of this study indicate that [(11)C]NPA can be used to measure D(2/3) receptors configured in a state of high affinity for the agonists with high reliability and reproducibility in the functional subdivisions of the human striatum. PMID- 19301418 TI - Ordinal and cardinal measures of health inequality: an empirical comparison. AB - When measuring health inequality using ordinal data, analysts typically must choose between indices specifically based upon ordinal data and more standard indices using ordinal data, which has been transformed into cardinal data. This paper compares inequality rankings across a number of different approaches and finds considerable sensitivity to the choice between ordinal- and cardinal-based indices. There is relatively little sensitivity to the ethical choices made by the analyst in terms of the weight attached to different parts of the distribution. PMID- 19301417 TI - Changes in mouse mu opioid receptor Exon 7/8-like immunoreactivity following food restriction and food deprivation in rats. AB - Opioid agonists and antagonists respectively increase and decrease food intake. That selective mu opioid antagonists are more effective than antisense probes directed against the mu opioid receptor (MOR-1) gene in reducing deprivation induced feeding suggests a role for isoforms. Both food restriction and deprivation alter protein and mRNA levels of opioid peptides and receptors. Antisera directed against Exon 4 of the MOR-1-like immunoreactivity (LI) (Exon 4) clone or directed against mouse Exons 7/8 (mE7/8-LI) revealed high levels of immunoreactivity in brain nuclei related to feeding behavior. Therefore, the present study assessed MOR-1LI and mE7/8-LI in hypothalamic and extrahypothalamic sites in rats exposed to ad libitum feeding, food restriction (2, 7, 14 days), or food deprivation (24, 48 h). MOR-1-LI displayed robust reactivity, but was insensitive to food restriction or deprivation. mE7/8-LI, both in terms of cell counts and relative optical density, was significantly and selectively increased in the dorsal and ventral parvocellular subdivisions of the hypothalamic paraventricular nucleus in food-restricted (14 days) rats, but all other restriction or deprivation regimens were ineffective in other hypothalamic nuclei. In contrast, significant and site-specific decreases in relative optical density in the rostral part of the nucleus tractus solitarius (NTS) were observed in food-restricted (2, 7 days) or food-deprived (24, 48 h) animals, but these regimens were ineffective in the other extrahypothalamic sites. This study indicates the sensitivity of this mE7/8-LI probe in the hypothalamic parvocellular paraventricular nucleus and rostral NTS to food restriction and deprivation in rats. PMID- 19301419 TI - A thirteen-year follow-up study of young Norwegian adults with dyslexia in childhood: reading development and educational levels. AB - As part of a larger follow-up study, 75 students were diagnosed with dyslexia at 10 years of age. At the end of secondary school, grades for the whole cohort of 16-year olds were examined. At age 23 the whole study group responded to a questionnaire and the subgroup with dyslexia who responded to this questionnaire were invited to come for testing and interviews. The persistence of dyslexia into young adulthood and educational levels were examined. The results showed that almost all students still suffered dyslexia and showed lower decoding ability. However, they only showed slightly lower educational attainment levels compared with a representative sample of half the cohort (n=530) they were part of. PMID- 19301420 TI - Concentration and drug prices in the retail market for malaria treatment in rural Tanzania. AB - The impact of market concentration has been little studied in markets for ambulatory care in the developing world, where the retail sector often accounts for a high proportion of treatments. This study begins to address this gap through an analysis of the consumer market for malaria treatment in rural areas of three districts in Tanzania. We developed methods for investigating market definition, sales volumes and concentration, and used these to explore the relationship between antimalarial retail prices and competition.The market was strongly geographically segmented and highly concentrated in terms of antimalarial sales. Antimalarial prices were positively associated with market concentration. High antimalarial prices were likely to be an important factor in the low proportion of care-seekers obtaining appropriate treatment.Retail sector distribution of subsidised antimalarials has been proposed to increase the coverage of effective treatment, but this analysis indicates that local market power may prevent such subsidies from being passed on to rural customers. Policymakers should consider the potential to maintain lower retail prices by decreasing concentration among antimalarial providers and recommending retail price levels. PMID- 19301421 TI - Biochemical analysis of oligomerization of expanded polyalanine repeat proteins. AB - Many human proteins contain amino acid repeats that can form homopolymeric amino acid (HPAA) tracts. HPAA tract proteins that contain polyalanine sequences promote diseases, including oculopharyngeal muscular dystrophy. The pathological properties of these proteins develop when the repeats match or exceed approximately 20 residues. We analyzed the oligomerization of yellow fluorescent protein (YFP) and GST fusion proteins containing >20 alanine repeats by using sucrose density gradient centrifugation. YFP and GST fusion proteins having 23 polyalanine residues sedimented readily in sucrose density gradients, suggesting instability and oligomerization of proteins with an excess of 20 alanine repeats. Moreover, GST fusion proteins were resistant to trypsin digestion after oligomerization. Oligomerized artificial proteins with long polyalanine repeats may be suitable models for studying polyalanine-related diseases. PMID- 19301422 TI - Neurotrophic effect of gonadotropin-releasing hormone on neurite extension and neuronal migration of embryonic gonadotropin-releasing hormone neurons in chick olfactory nerve bundle culture. AB - Hypothalamic gonadotropin-releasing hormone (GnRH) neurons play a pivotal role in regulating the reproductive function of vertebrates. These neurons are known to originate in the olfactory placode and migrate along olfactory-related axons to reach the forebrain during embryonic development. Although GnRH is suggested to be secreted during such migration, its physiological significance is unknown. This point is difficult to explore in vivo because recent studies suggest that GnRH is an important factor for normal brain development and that modification of the embryonic GnRH system by exogenous GnRH analogue or genetic methods would result in dysgenesis of the brain. Therefore, to study the role of GnRH in the migratory process of GnRH neurons, we established an in vitro chick embryonic olfactory nerve bundle explant model. Embryonic day 7.5-8 olfactory nerve bundles were cultured in a mixture of Matrigel and collagen gel. At day 3 of culture, GnRH neurons extended their unbranched neurites and migrated out from both edges of the explant. The nature of neurite extension and migratory behavior of GnRH neurons was well maintained in the gel containing 25% Matrigel and 50% collagen. With this culture system, we examined the effect of GnRH on the migrating GnRH neurons. Cetrorelix, a GnRH antagonist, was found to inhibit significantly neurite growth and neuronal migration of GnRH neurons, the effects of which were repressed by the addition of chicken GnRH-I. These results suggest that GnRH functions as one of the regulating factors of GnRH neuronal development by promoting neurite extension and neuronal migration. PMID- 19301423 TI - Soluble mediators from human neural stem cells play a critical role in suppression of T-cell activation and proliferation. AB - Human neural stem cells (hNSCs) can control inflammation in the central nervous system, although the underlying mechanisms are not understood fully. We investigated the immunomodulatory effect of hNSCs on human T cells and the underlying mechanisms. Culture supernatant from an immortalized hNSC cell line, HB1.F3, which has a therapeutic effect on acute stroke and intracerebral hemorrhage, suppressed the proliferation of allogeneically or mitogenically stimulated human peripheral T cells, including the CD3(+)CD103(+) subpopulation. CFSE labeling and flow cytometry showed that the suppression of proliferation was caused by cell cycle arrest and induction of apoptosis. The lack of significant change in caspase-8 levels and the significant reduction in Bcl-2 expression in the affected T cells suggest that the intrinsic pathway plays a major role in soluble-factor-mediated T-cell apoptosis. The addition of culture supernatant from hNSCs to activated T cells reduced the expression of the activation markers CD69 and CD25 at 24 hr after activation, but at 48 hr only CD69 was down regulated. A cytometry bead assay showed that the secretion of interleukin (IL)-2 decreased significantly, whereas that of IL-4, IL-10, tumor necrosis factor alpha, and interferon-gamma increased. These results show that hNSCs can negatively affect human peripheral T cells by suppressing their activation and proliferation through soluble mediators, suggesting that hNSCs have a bystander immunomodulatory effect on T cells. PMID- 19301424 TI - Neuroprotective role of erythropoietin by antiapoptosis in the retina. AB - Erythropoietin (EPO) stimulates red blood cell production, in part by inhibiting apoptosis of the red blood cell precursors. The erythropoietic effects of EPO are circadian stage dependent. Retinal injury due to light occurs through oxidative mechanisms and is manifest by retinal and retinal pigment epithelium (RPE) cells apoptosis. The visual cycle might be circadian coordinated as a means of effectively protecting the retina from the detrimental effects of light-induced, oxygen-dependent, free radical-mediated damage, especially at the times of day when light is more intense. We show that the retinal expression of EPO and its receptor (EPOR), as well as subsequent Janus kinase 2 (Jak2) phosphorylations, are each tightly linked to a specific time after oxidative stress and in anticipation of daily light onset. This is consistent with physiological protection against daily light-induced, oxidatively mediated retinal apoptosis. In vitro, we verify that EPO protects RPE cells from light, hyperoxia, and hydrogen peroxide-induced retinal cell apoptosis, and that these stimuli increase EPO and EPOR expression in cultured RPE cells. Together, these data support the premise that EPO and its EPOR interactions represent an important retinal shield from physiologic and pathologic light-induced oxidative injury. PMID- 19301425 TI - N-cadherin mediates interaction between precursor cells in the subventricular zone and regulates further differentiation. AB - Neurogenesis and cell differentiation in the brain continues throughout life. In the subventricular zone and rostral migratory stream, precursor cells contact each other. Cell-cell interactions mediated via adhesion molecules are no doubt involved in establishing and maintaining the neurogenic ability of these cells. Here, we demonstrate that N-cadherin plays important roles in forming cell clusters and in regulating cell differentiation. N-cadherin is abundantly expressed in chain migrating cells in the subventricular zone and rostral migratory stream but is down-regulated after cells exit these regions. We also show that neurosphere formation is inhibited via suppression of N-cadherin function and that N-cadherin expression is decreased after induction of neurosphere differentiation. Furthermore, we demonstrate that functional blockade of N-cadherin can enhance glial cell differentiation in explant cultures of precursors from the subventricular zone. PMID- 19301426 TI - Human synapsin I mediates the function of nuclear respiratory factor 1 in neurite outgrowth in neuroblastoma IMR-32 cells. AB - Nuclear respiratory factor (NRF)-1 is a transcription factor with a novel function in neurite outgrowth. Synapsin I protein is a well-known phosphoprotein in neuronal terminals and has been implicated in neuronal differentiation. Human synapsin I gene promoter has a putative NRF-1 responsive element (NRE), but it is not known whether this NRE is functional. We hypothesized that synapsin I is downstream of NRF-1 and mediates its function in neurite outgrowth. Gel electrophoretic mobility shift assays, chromatin immunoprecipitation, site directed mutagenesis, and promoter studies indicated that NRF-1 is a positive regulator of synapsin I promoter. Exogenous NRF-1 overexpression increased synapsin I protein levels in IMR-32 and HEK293T cells. Serum deprivation, which induces neurite outgrowth in IMR-32 cells, increased the binding activity of NRF 1 to synapsin I NRE and induced alternating synapsin I protein expression. Down regulating synapsin I expression markedly decreased the percentage of neurite bearing cells and the length of the longest neurite in IMR-32 cells that stably or transiently overexpressed NRF-1. We conclude that the human synapsin I gene is positively regulated by NRF-1 and mediates the function of NRF-1 in neurite outgrowth. PMID- 19301428 TI - Amyloid-beta interrupts the PI3K-Akt-mTOR signaling pathway that could be involved in brain-derived neurotrophic factor-induced Arc expression in rat cortical neurons. AB - The deposition of amyloid-beta (Abeta) contributes to the pathogenesis of Alzheimer's disease. Even at low levels, Abeta may interfere with various signaling cascades critical for the synaptic plasticity that underlies learning and memory. Brain-derived neurotrophic factor (BDNF) is well known to be capable of inducing the synthesis of activity-regulated cytoskeleton-associated protein (Arc), which plays a fundamental role in modulating synaptic plasticity. Our recent study has demonstrated that treatment of fibrillar Abeta at a nonlethal level was sufficient to impair BDNF-induced Arc expression in cultured rat cortical neurons. In this study, BDNF treatment alone induced the activation of the phosphatidylinositol 3-kinase-Akt-mammlian target of rapamycin (PI3K-Akt mTOR) signaling pathway, the phosphorylation of eukaryotic initiation factor 4E binding protein (4EBP1) and p70 ribosomal S6 kinase (p70S6K), the dephosphorylation of eukaryotic elongation factor 2 (eEF2), and the expression of Arc. Interrupting the PI3K-Akt-mTOR signaling pathway by inhibitors prevented the effects of BDNF, indicating the involvement of this pathway in BDNF-induced 4EBP1 phosphorylation, p70S6K phosphorylation, eEF2 dephosphorylation, and Arc expression. Nonlethal Abeta pretreatment partially blocked these effects of BDNF. Double- immunofluorescent staining in rat cortical neurons further confirmed the coexistence of eEF2 dephosphorylation and Arc expression following BDNF treatment regardless of the presence of Abeta. These results reveal that, in cultured rat cortical neurons, Abeta interrupts the PI3K-Akt-mTOR signaling pathway that could be involved in BDNF-induced Arc expression. Moreover, this study also provides the first evidence that there is a close correlation between BDNF-induced eEF2 dephosphorylation and BDNF-induced Arc expression. (c) 2009 Wiley-Liss, Inc. PMID- 19301427 TI - Distinct modes of migration position oligodendrocyte precursors for localized cell division in the developing spinal cord. AB - Establishment of the cytoarchitecture of the central nervous system reflects the stereotyped cell migration and proliferation of precursor cells during development. In vitro analyses have provided extensive information on the control of proliferation and differentiation of oligodendrocyte precursors (OPCs), but less is known about the migratory behavior of these cells in vivo. Here we utilize a transgenic mouse line expressing enhanced green fluorescent protein (EGFP) under the proteolipid protein promoter (PLP-EGFP mice) to visualize directly the behaviors of OPCs in developing spinal cord slices. During early development, OPCs disperse from their origin at the ventricular zone by using saltatory migration. This involves orientation of the cell with a leading edge toward the pial surface and alternating stationary and fast-moving phases and dramatic shape changes. Once cells exit the ventricular zone, they exhibit an exploratory mode of migration characterized by persistent translocation without dramatic changes in cell morphology. The control of migration, proliferation, and cytokinesis of OPCs appear to be closely linked. In netrin-1 mutant spinal cords that lack dispersal cues, OPC migration rates were not significantly different, but the trajectories were altered, and numbers of migrating cells were dramatically reduced. In contrast to DNA replication that occurs at the ventricular zone or throughout the spinal cord neuropil, cell division or cytokinesis of OPCs occurs predominantly at the interface between gray and white matters, with the majority of cleavage planes parallel to the pial surface. These studies suggest that positional cues are critical for regulating OPC behavior during spinal cord development. PMID- 19301429 TI - Density of cerebellar basket and stellate cells in autism: evidence for a late developmental loss of Purkinje cells. AB - Alterations in the cerebellum have been described as a neuropathological feature of autism. Although numerous studies have focused on the Purkinje cell (PC), the projection neuron of the cerebellar cortex, PC function is critically dependent on their innervation by the GABAergic basket cells (BCs) and stellate cells (SCs) in the cerebellar molecular layer. The present study was designed to determine whether there are differences in the packing density of these inhibitory interneurons or whether the ratio of these interneurons to PCs differs in autistic and age-matched control brains. The GABAergic interneurons were identified by using immunohistochemistry for parvalbumin (PV) in serial sections from the posterior cerebellar lobe of six autistic and four control brains and counted using stereological principles. Prior PC counts in the same area on adjacent sections (Whitney et al., 2008) were available and were used to calculate the number of BCs and SCs per PC. In this sample of brains, no statistically significant difference was detected between the autistic and the control groups in the density of BCs or SCs (P = 0.44 and P = 0.84, respectively) or in the number of BCs or SCs per PC (P = 0.47 and P = 0.44, respectively). The preservation of BCs and SCs, in the presence of the reduced PC numbers as found in at least two, and possibly three, of these six autistic cases (Whitney et al., 2008) suggests that PCs were generated, migrated to their proper location in the PC layer, and subsequently died in the autistic cases that showed a reduction in PCs. PMID- 19301430 TI - gamma-Aminobutyric acid-mediated regulation of the activity-dependent olfactory bulb dopaminergic phenotype. AB - gamma-Aminobutyric acid (GABA) regulates the proliferation and migration of olfactory bulb (OB) interneuron progenitors derived from the subventricular zone (SVZ), but the role of GABA in the differentiation of these progenitors has been largely unexplored. This study examines the role of GABA in the differentiation of OB dopaminergic interneurons using neonatal forebrain organotypic slice cultures prepared from transgenic mice expressing green fluorescent protein (GFP) under the control of the tyrosine hydroxylase (Th) gene promoter (ThGFP). KCl mediated depolarization of the slices induced ThGFP expression. The addition of GABA to the depolarized slices further increased GFP fluorescence by inducing ThGFP expression in an additional set of periglomerular cells. These findings show that GABA promoted differentiation of SVZ-derived OB dopaminergic interneurons and suggest that GABA indirectly regulated Th expression and OB dopaminergic neuron differentiation through an acceleration of the maturation rate for the dopaminergic progenitors. Additional studies revealed that the effect of GABA on ThGFP expression required activation of L- and P/Q-type Ca2+ channels as well as GABA(A) and GABA(B) receptors. These voltage-gated Ca2+ channels and GABA receptors have previously been shown to be required for the coexpressed GABAergic phenotype in the OB interneurons. Together, these findings suggest that Th expression and the differentiation of OB dopaminergic interneurons are coupled to the coexpressed GABAergic phenotype and demonstrate a novel role for GABA in neurogenesis. PMID- 19301431 TI - Stem cell-based cell therapy in neurological diseases: a review. AB - Human neurological disorders such as Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), Alzheimer's disease, multiple sclerosis (MS), stroke, and spinal cord injury are caused by a loss of neurons and glial cells in the brain or spinal cord. Cell replacement therapy and gene transfer to the diseased or injured brain have provided the basis for the development of potentially powerful new therapeutic strategies for a broad spectrum of human neurological diseases. However, the paucity of suitable cell types for cell replacement therapy in patients suffering from neurological disorders has hampered the development of this promising therapeutic approach. In recent years, neurons and glial cells have successfully been generated from stem cells such as embryonic stem cells, mesenchymal stem cells, and neural stem cells, and extensive efforts by investigators to develop stem cell-based brain transplantation therapies have been carried out. We review here notable experimental and preclinical studies previously published involving stem cell based cell and gene therapies for Parkinson's disease, Huntington's disease, ALS, Alzheimer's disease, MS, stroke, spinal cord injury, brain tumor, and lysosomal storage diseases and discuss the future prospects for stem cell therapy of neurological disorders in the clinical setting. There are still many obstacles to be overcome before clinical application of cell therapy in neurological disease patients is adopted: 1) it is still uncertain what kind of stem cells would be an ideal source for cellular grafts, and 2) the mechanism by which transplantation of stem cells leads to an enhanced functional recovery and structural reorganization must to be better understood. Steady and solid progress in stem cell research in both basic and preclinical settings should support the hope for development of stem cell-based cell therapies for neurological diseases. PMID- 19301432 TI - Corticospinal neurons respond differentially to neurotrophins and myelin associated glycoprotein in vitro. AB - Elucidating the mechanisms that regulate the survival and outgrowth of corticospinal tract (CST) neurons and other CNS tracts will be a key component in developing novel approaches for the treatment of central nervous system (CNS) disorders, including stroke, spinal cord injury (SCI), and motor neuron disease (MND). However, the in vivo complexities of these diseases make a systematic evaluation of potential therapeutics that directly affect corticospinal regeneration or survival very challenging. Here, we use Thy1.2 transgenic mice expressing yellow fluorescent protein (YFP) in postnatal day 8 (P8) corticospinal neurons, as a source of CST neurons that have already established synapses in the spinal cord, to assess factors that influence neurite outgrowth and survival of axotomized CST neurons. After culture, YFP-positive corticospinal neurons represent an enriched neuronal population over other glia and interneurons, survive, and extend processes over time. YFP-positive CST neurons also continue to express the corticospinal markers CTIP2 and Otx1. CST neurons display different degrees of axon extension, dendritic branch length and elaboration, and neurite elongation in response to neurotrophin-3 and ciliary neurotrophic factor, and an inhibitory outgrowth response when cultured on myelin-associated glycoprotein. Some CST neurons are lost with extended culture, which provides a baseline from which we can also assess factors that enhance CST neuron survival. This assay thus allows us to assess independent aspects of CST axonal and dendritic outgrowth kinetics, which allows for the rapid and sensitive investigation of new therapies to address corticospinal neuron outgrowth in the context of CNS injury and neurodegenerative disorders. PMID- 19301433 TI - NCI's Community Cancer Center Program shows progress. PMID- 19301434 TI - Benefit of postoperative adjuvant therapy for pancreatic cancer: A population based analysis. AB - BACKGROUND: Despite the recent completion of several trials of adjuvant therapy after resection for pancreatic adenocarcinoma, the absolute impact on survival and the identification of appropriate patients for treatment has remained controversial. In the current study, the authors sought to identify the impact of adjuvant therapy and factors associated with any improvement in survival after resection of pancreatic cancer. METHODS: Through the California Cancer Registry, all California residents diagnosed with pancreatic cancer between 1994 and 2002 were identified. Factors potentially impacting survival were analyzed, including patient demographics, tumor characteristics, and treatment provided. Univariate and multivariate survival analyses were performed by Kaplan-Meier and Cox regression methods. RESULTS: A total of 26,518 patients were identified; 3196 (12.1%) underwent resection as their primary treatment. The median overall survival was 16 months for patients who underwent resection. Prognostic factors associated with better survival included negative lymph node status, well differentiated tumors, younger age, female sex, and the receipt of any adjuvant therapy. On multivariate analysis, adjuvant therapy demonstrated a statistically significant, although modest, impact on survival, with a hazards ratio of 0.79 (95% confidence interval, 0.72-0.87; P < .001). The benefit of adjuvant therapy was only apparent in those patients with lymph node-positive or poorly differentiated tumors. CONCLUSIONS: Adjuvant therapy provided for a modest improvement in overall survival after surgical resection of pancreatic cancer. The absolute effect was most pronounced in those patients with poor prognostic indicators. To identify effective systemic therapy for this deadly cancer, future clinical trials of adjuvant therapy should focus on these groups of patients. PMID- 19301435 TI - Pharmaceuticals and medical devices: Medicare Part D. End-of-year issue brief. PMID- 19301439 TI - HIV-positive woman acquitted of aggravated sexual assault. PMID- 19301440 TI - Halifax man convicted of aggravated sexual assault for failure to disclose HIV-positive status. PMID- 19301441 TI - Man sentenced to five-year prison term for failing to disclose his HIV positive status. PMID- 19301442 TI - Man sentenced to seven years imprisonment on eight counts of aggravated sexual assault. PMID- 19301443 TI - Singapore: man convicted for performing oral sex. PMID- 19301444 TI - HIV-positive Kenyan awarded compensation after being fired. PMID- 19301445 TI - China: judicial interpretation imposes stricter responsibility for contaminated blood sales. PMID- 19301446 TI - Brazilian Appeals Court rules that criminal prosecution for drug possession is unconstitutional. PMID- 19301447 TI - New format, open access, and online pre-publication. PMID- 19301448 TI - Irbesartan for heart failure with preserved ejection fraction. PMID- 19301449 TI - Irbesartan for heart failure with preserved ejection fraction. PMID- 19301450 TI - Irbesartan for heart failure with preserved ejection fraction. PMID- 19301451 TI - Compliance with azithromycin versus erythromycin in the setting of a pertussis outbreak. AB - BACKGROUND: Erythromycin has traditionally been the choice for prophylaxis and treatment of pertussis, but recently azithromycin has been recommended as another first-line agent. We evaluated treatment adherence between exposed persons giving erythromycin or azithromycin during a community-wide pertussis outbreak. METHODS: This was a case-control study. All cases and their contacts were prescribed either 56 doses of erythromycin over 14 days or 5 doses of azithromycin over 5 days. A standardized questionnaire regarding demographics, side effects, and compliance with therapy was administered by mail or telephone interviews. RESULTS: Of 244 persons prescribed erythromycin, 139 (57%) completed the full course compared with 234 (93%) of 251 persons prescribed azithromycin (rate ratio [RR] 4.5; 95% confidence interval [CI], 2.9-7.0). The primary reason for not completing erythromycin was side effects in 79 (76%) persons, of whom 72 (91%) reported gastrointestinal upset, compared with azithromycin side effects in 6 (35%) of whom 5 (83%) reported gastrointestinal side effects. CONCLUSIONS: Azithromycin was associated with significantly higher completion rates than erythromycin. Due to side effects, the use of azithromycin may be preferable to erythromycin in outbreaks of pertussis. PMID- 19301452 TI - Transcatheter interatrial shunt closure as a cure for migraine: can it be justified by paradoxical embolism-risk-driven criteria? AB - BACKGROUND: Some ongoing trials have suggested that closure of the patent foramen ovale (PFO) may reduce migraine symptoms. We sought to assess the safety and effectiveness of migraine treatment by means of PFO transcatheter closure using paradoxical embolism risk-driven criteria. METHODS: We enrolled 75 patients (48 women and 27 men, mean age 40 +/- 3.7 years) who were referred to our center over a 12-month period for a prospective study to evaluate severe disabling migraine, despite antiheadache therapy and the PFO. Migraine Disability Assessment Score (MIDAS) was used to assess the incidence of migraine headache and severity. Criteria for intervention included all of the following: basal shunt, curtain shunt pattern on transcranial Doppler, presence of interatrial septal aneurysm, 3 to 4 class MIDAS score, symptomatic significant aura, coagulation abnormalities, migraine refractory to conventional drugs. RESULTS: On the basis of the inclusion criteria, we shortlisted 20 patients (12 women, mean age 35 +/- 6.7 years, mean MIDAS score 38.9 +/- 5.8) for transcatheter closure of PFO and excluded the rest who were referred to the neurologist for medical therapy. The procedure was successful in all of the patients with no perioperative or in-hospital complications. After a mean follow-up of 10 +/- 3.1 months (range 6-14), all patients' migraine symptoms improved (mean MIDAS score 3.0 +/- 2.1, P < 0.03) with PFO complete closure in all patients on transesophageal and transcranial Doppler ultrasound. CONCLUSION: In this small pilot series, we adopted the criteria which in our opinion best reflected the risk of paradoxical embolism in these patients. By adopting the proposed criteria, primary transcatheter closure of the PFO resulted in a significant reduction in migraine. PMID- 19301453 TI - Nontuberculous mycobacteria among patients who are suspected for multidrug resistant tuberculosis-need for earlier identification of nontuberculosis mycobacteria. AB - BACKGROUND: In this study, we intended to find the prevalence of nontuberculosis mycobacteria (NTM) among patients who are referred as suspected multidrug resistant tuberculosis (MDR-TB) cases to the only referral center in Iran. METHODS: All patients referred to our center in 2002-2006 as MDR-TB with histories of treatment with standard and CAT II World Health Organization regimens were included in the study. Sputum smear and culture for acid-fast bacilli were performed for all patients 3 times. Sputum polymerase chain reaction was also performed for all patients. Mycobacterial identification was performed via polymerase chain reaction and routine identification tests for all culture positive cases. RESULTS: Of the 105 patients in the study, 12 (11.43%) were identified to have NTM infection. The identified mycobacteria were classified in order of prevalence as Chelonae (8 cases), Simiae (2 cases), Aloei (1 case), and Farcinogen (1 case). Based on radiologic findings, most of the cases demonstrated bilateral nodularity (83.3%) and also multifocal bronchiectasis (75%). Notably, cavitary lesions were present in 41.7% of the cases. CONCLUSION: Based on the findings of this study, it is essential that such cases be identified before commencing MDR-TB treatment. PMID- 19301454 TI - Does reporting estimated glomerular filtration rate affect ordering of timed urine collections? AB - BACKGROUND: The National Kidney Foundation recommends reporting estimates of glomerular filtration rate (eGFR) rather than timed urine collections. When 2 of the 3 major hospitals in our region began reporting eGFR at different times we recognized a natural experiment. METHODS: We conducted a retrospective, observational study at the 3 major hospitals in Chattanooga, Tennessee. Data were collected on the frequency of timed urines during a 41/2-year period. Regression analysis was used to study the association of the rate of ordering timed urines adjusted for other factors. RESULTS: There was a marked drop in the rate of ordering of timed urines at all 3 hospitals from a mean of 21.8 per 1000 admissions in the first 4 quarter years of the study to 10.9 in the last 4 (15th 18th) quarter years. The drop began before the reporting of eGFR at any hospital. The reporting of eGFR had a small effect (-1.7 per 1000 hospital admissions) that was not statistically significant (P = 0.15). CONCLUSIONS: There has been a marked drop in the ordering of timed urines in our region. The decline began before the reporting of eGFR at 2 of the hospitals and therefore is attributable to other factors. PMID- 19301455 TI - Antiglomerular basement membrane disease associated with Castleman disease. AB - We described a 33-year-old man with gross hematuria and acute renal failure, who had suffered from fever for 6 months, multiple cervical lymph node swelling, splenomegaly, left-sided pleural effusion. He also suffered from anemia, thrombocytopenia, hypergammaglobulinemia, and his serum interleukin-6 levels were markedly elevated. Antiglomerular basement membrane antibodies were positive in the patient's serum. Lymph node biopsy results were compatible with Castleman disease of "plasma cell" variant. Renal biopsy revealed cellular crescents in most of the glomeruli. Immunofluorescence studies showed strong deposition of IgG in a linear pattern along the glomerular basement membrane. Pathologic features were compatible with crescentic glomerulonephritis because of antiglomerular basement membrane disease. With intensive plasmapheresis and monthly chemotherapy (cyclophosphamide, oncovin, prednisone regimen), the patient experienced clinical and biochemical remission. Although autoimmune phenomenon had been described frequently in Castleman disease, to the best of our knowledge, this was the first report that the patient with rapid progressive glomerulonephritis mediated by antiglomerular basement membrane antibodies, which might be associated with Castleman disease. PMID- 19301456 TI - Stereotactic radiosurgery for trigeminal schwannoma: tumor control and functional preservation Clinical article. AB - OBJECT: To evaluate outcome predictors after stereotactic radiosurgery (SRS) in patients with trigeminal schwannomas, the authors compared tumor control, functional preservation, and complications with tumor grade, tumor volume, patient age, and tumor imaging characteristics. METHODS: The records of 33 consecutive patients with trigeminal schwannoma treated via Gamma Knife surgery were retrospectively reviewed. The median patient age was 49.5 years (range 15.1 82.5 years). Eleven patients had undergone prior tumor resection. Two patients had neurofibromatosis Type 2. Lesions were classified as root type (6 tumors), ganglion type (17 tumors), and dumbbell type (10 tumors) based on their location. The median radiosurgery target volume was 4.2 cm3 (range 0.5-18.0 cm3), and the median dose to the tumor margin was 15.0 Gy (range 12-20 Gy). RESULTS: At an average of 6 years (range 7.2-147.9 months), the rate of progression-free survival (PFS) at 1, 5, and 10 years after SRS was 97.0, 82.0, and 82.0%, respectively. Factors associated with improved PFS included female sex, smaller tumor volume, and a root or ganglion tumor type. Neurological symptoms or signs improved in 11 (33.3%) of 33 patients and were unchanged in 19 (57.6%). Three patients (9.1%) had symptomatic disease progression. Patients who had not undergone a prior tumor resection were significantly more likely to show improvement in neurological symptoms or signs. CONCLUSIONS: Stereotactic radiosurgery is an effective and minimally invasive management option in patients with residual or newly diagnosed trigeminal schwannomas. Predictors of a better treatment response included female sex, smaller tumor volume, root or ganglion tumor type, and the application of SRS as the primary treatment. PMID- 19301457 TI - MUM1 expression does not differentiate primary cutaneous anaplastic large-cell lymphoma and lymphomatoid papulosis. PMID- 19301458 TI - Early initiation of ART produces better clinical outcomes. PMID- 19301459 TI - IL-2 therapy provides no clinical benefit to HIV-infected patients on ART. PMID- 19301460 TI - AIDS top killer disease in China last year: govt. PMID- 19301461 TI - Bibliography. Current world literature. Cardiovascular anaesthesia. PMID- 19301462 TI - The use of bioabsorbable screw in a split anterior tibial tendon transfer: a preliminary result. AB - The split anterior tibial tendon transfer is a procedure commonly used to correct equinovarus deformity of the foot, primarily in children with spastic cerebral palsy. The procedure has been reported to yield satisfactory results; it can be combined with other soft tissue lengthening. Typically, the transferred tendon can be fixed by either button, staple, metal screw, or anchored into two bony tunnels. Occasionally, failure to maintain transferred tendon fixation may result from conventional surgical methods and therefore lead to undesirable results. The technique of using bioabsorbable interference screw has been proved to be effective in securing the graft as anterior cruciate ligament reconstruction in knee joint. To our knowledge, only few literatures have reported on the use of bioabsorbable screw in the treatment of tendon transfers around the foot. From 2004 to 2006, split anterior tibial tendon transfers with concomitant tendo Achilles lengthening were applied to 13 patients (16 feet) who had a dynamic equinovarus deformity secondary to spastic cerebral palsy. Mean age at surgery was 8 years and the average follow-up period was 16 months (range, 12-28 months). A 7 x 23 mm cannulated bioabsorbable interference screw was used to fix the transferred tendon. Postoperatively, we used the rating system of Kling et al. to assess the clinical results. Our findings showed that there were 10 excellent, six good, and no poor results. All patients had improved gait at their latest follow-ups. At final follow-up, there was no graft failures or postoperative complications related to the bioabsorbable screw in our series. Although the study is based on a small number of cases, we suggest this technique is an additional armamentarium in fixation for tendon transfers in children's feet. PMID- 19301463 TI - Clinical protocol number #19: breastfeeding promotion in the prenatal setting. AB - A central goal of The Academy of Breastfeeding Medicine is the development of clinical protocols for managing common medical problems that may impact breastfeeding success. These protocols serve only as guidelines for the care of breastfeeding mothers and infants and do not delineate an exclusive course of treatment or serve as standards of medical care. Variations in treatment may be appropriate according to the needs of an individual patient. PMID- 19301464 TI - Clopidogrel pretreatment before primary percutaneous coronary stenting in patients with acute ST-segment elevation myocardial infarction: comparison of high loading dose (600 mg) versus low loading dose (300 mg). AB - BACKGROUND: Aggressive platelet inhibition is crucial to reduce myocardial injury and early cardiac events after coronary intervention. As compared with the conventional 300-mg dose, pretreatment with a 600-mg loading dose of clopidogrel significantly reduced periprocedural myocardial infarction (MI) in patients undergoing percutaneous coronary intervention (PCI). We investigated that the advantage of the 600-mg dose in inhibiting platelet aggregation more rapidly than the 300-mg dose may actually have special value for acute ST-segment elevation MI patients. METHODS: A total of 171 patients with ST-segment elevation MI underwent primary PCI. A 600-mg (n=73) or 300-mg (n=98) loading regimen of clopidogrel was given before the procedure. We did a follow-up of all patients clinically for 30 days after coronary intervention. The primary endpoint was the 30-day occurrence of death, MI, urgent revascularization, or stroke. RESULTS: The primary endpoint occurred in 1.4% (1 of 73) of patients in the high dose versus 11.2% (11 of 98) of those in the conventional loading dose group (P=0.013). Death, recurrent MI, urgent revascularization, and stroke were lower in patients treated with the high dose of clopidogrel compared with conventional dose. Safety endpoints were similar in the two groups. CONCLUSION: Pretreatment with a 600-mg loading dose of clopidogrel before the procedure is safe and, as compared with the conventional 300-mg dose, significantly reduces recurrent MI and urgent revascularization in patients with primary PCI. PMID- 19301465 TI - Update: the Joint Commission's hospital deeming authority application. PMID- 19301466 TI - Errata: 2009 accreditation manuals, all programs. PMID- 19301467 TI - Correction: health care staffing services certification criticality designations. PMID- 19301468 TI - CMS continues to recognize Joint Commission ambulatory surgical center accreditation. PMID- 19301469 TI - Advance trauma life support refresher course encouraged. PMID- 19301470 TI - Metal-free cascade intramolecular S-arylation: regioselective synthesis of substituted benzothiazoles. PMID- 19301471 TI - Optimization of intradermal vaccination by DNA tattooing in human skin. AB - The intradermal administration of DNA vaccines by tattooing is a promising delivery technique for genetic immunization, with proven high immunogenicity in mice and in nonhuman primates. However, the parameters that result in optimal expression of DNA vaccines that are applied by this strategy to human skin are currently unknown. To address this issue we set up an ex vivo human skin model in which DNA vaccine-induced expression of reporter proteins could be monitored longitudinally. Using this model we demonstrate the following: First, the vast majority of cells that express DNA vaccine-encoded antigen in human skin are formed by epidermal keratinocytes, with only a small fraction (about 1%) of antigen-positive epidermal Langerhans cells. Second, using full randomization of DNA tattoo variables we show that an increase in DNA concentration,needle depth, and tattoo time all significantly increase antigen expression ( p < 0.001), with DNA concentration forming the most critical variable influencing the level of antigen expression. Finally, in spite of the marked immunogenicity of this vaccination method in animal models, transfection efficiency of the technique is shown to be extremely low, estimated at approximately 2 to 2000 out of 1 x 10(10) copies of plasmid applied. This finding, coupled with the observed dependency of antigen expression on DNA concentration, suggests that the development of strategies that can enhance in vivo transfection efficacy would be highly valuable. Collectively, this study shows that an ex vivo human skin model can be used to determine the factors that control vaccine-induced antigen expression and define the optimal parameters for the evaluation of DNA tattoo or other dermal delivery techniques in phase 1 clinical trials. PMID- 19301472 TI - Prolonged transgene expression with lentiviral vectors in the aqueous humor outflow pathway of nonhuman primates. AB - We injected lentiviral vectors into the eyes of live nonhuman primates to assess potential for glaucoma gene therapy. Anterior chambers of five cynomolgus monkeys were injected with green fluorescent protein (GFP)-encoding feline immunodeficiency viral vectors. The monkeys were monitored for in vivo transgene expression and clinical parameters. Their eyes were harvested 2-15 months postinjection for tissue analyses. All seven eyes injected with 1.0-2.0 x 10(8) transducing units (TU) showed substantial GFP fluorescence in the trabecular meshwork (TM), which was observable even by goniophotographic monitoring for up to 15 months. Only the lowest dose (0.03 x 10(8) TU) failed to result in TM fluorescence detectable in vivo, and five of the eight vector-injected eyes continued to display substantial GFP expression when enucleated eyes were examined at 2, 7, or 15 months postinjection. Some transduced cells were also detected in the iris and ciliary body. Mild, transient postinjection inflammatory responses exceeding that induced by a control saline injection were observed, but vectors did not raise intraocular pressure and were well tolerated. The results demonstrate the first lentiviral vector transduction of the nonhuman primate aqueous humor outflow pathway and support application of the system to human glaucoma gene therapy. PMID- 19301473 TI - Factors determining the risk of inadvertent retroviral transduction of male germ cells after in utero gene transfer in sheep. AB - The possibility of permanent genetic changes to the germline is central to the bioethics of in utero gene therapy (IUGT) because of the concern of inadvertent potentially deleterious alterations to the gene pool. Despite presumed protection of the male germline due to early germ cell (GC) compartmentalization, we reported that GCs within the developing ovine testes are transduced at low levels after retrovirus-mediated IUGT, thus underscoring the need for a thorough understanding of GC development in clinically predictive models to determine the optimal time to perform IUGT and avoid germline modification. In the present studies, we used the fetal sheep model to analyze GCs for phenotype, location, proliferation, and incidence of transduction after IUGT at various fetal ages to learn when during development the nascent germline is likely to be at greatest risk of retrovirus-mediated alteration. Our studies show that although GCs were transduced at all injection ages, the levels of transduction varied by nearly 700 fold as a function of the age at transfer. After remaining largely quiescent as they migrated to/settled within nascent sex cords, GCs began active cycling before cord closure was complete, suggesting this is likely the point at which they would be most susceptible to retroviral transduction.Furthermore, we observed that compartmentalization of GCs continued into early postnatal life, suggesting the male germline may be vulnerable to low-level inadvertent retroviral vector modification throughout fetal life, but that this risk can be minimized by performing IUGT later in gestation. PMID- 19301474 TI - Culturally informed smoking cessation strategies for Native Hawaiians. AB - We examined the perceived supports and barriers and the smoking cessation strategies used by Native Hawaiian former and current smokers for the purpose of developing a culturally informed smoking cessation program. Ten focus groups with a total of 52 Native Hawaiian men and women were convened in a rural community in Hawai'i. Thematic analysis of focus group transcriptions resulted in the identification of 11 strategies and 23 supports for and 13 barriers to smoking cessation that were categorized into social, psychological, physical, political, economic, behavioral, and spiritual factors. Native Hawaiian former smokers (compared with current smokers who had tried to quit) found social, psychological, and physical factors helpful in supporting smoking cessation and remaining smoke free. They also reported having used more behavioral and religious/spiritual strategies to quit smoking compared with current smokers. The stories of former smokers also spoke to the importance of family and their religion/spirituality in quitting. Consistent with the findings from other studies, multiple factors were implicated in smoking behavior, suggesting that a multicomponent strategy may be beneficial for addressing the social, psychological, and physical factors related to smoking that we observed in our sample of Native Hawaiians. We also recognize that religion/spirituality is an integral part of Native Hawaiian culture and that church-based support of smoking cessation may help those Native Hawaiians for whom religion/spirituality is an important source of inspiration and guidance. PMID- 19301475 TI - [Brain injury marker S100B can reduce the use of computer tomography in minor head injuries--secondary publication]. AB - The risk of acute intracranial complications after minor head injury (MHI) is low. Despite this, computed tomography (CT) is generally recommended with clinical observation as a secondary option. Both options have disadvantages. Clinical studies have shown the potential advantages of using the biomarker S100B. The specificity of S100B is low, but a high sensitivity for brain damage results in a clinically useful, high negative predictive value (NPV). Integration of S100B into existing management routines can reduce the need for CT scans or admission by over 30%. PMID- 19301476 TI - [The effect of length of stay and number of relocations on asylum-seeking children's mental health--secondary publication]. AB - Among asylum-seeking children aged 4-16 years living in the asylum centres managed by the Danish Red Cross in 2006, we investigated mental health in relation to length of stay and number of relocations. Mental health was evaluated using the Strengths and Difficulties Questionnaire (SDQ). We found that children who had been asylum-seeking for more than one year had an increased risk of having mental difficulties (odds ratio 5.5; 95% confidence interval 1.8-16.3); four or more relocations in the asylum system were also associated with a higher risk of mental problems (odds ratio 3,0; 1,4-6,7). PMID- 19301477 TI - [That is why your article was rejected]. PMID- 19301478 TI - [DAMP on the right track?]. PMID- 19301479 TI - [Quality control in general practice]. PMID- 19301480 TI - Investing in healthy babies. PMID- 19301481 TI - Tracking cancer screening progress. PMID- 19301483 TI - BVA president emphasises importance of biosecurity for all species. PMID- 19301482 TI - Disparities in cardiovascular health. PMID- 19301484 TI - Physical activity guidelines. PMID- 19301486 TI - Communication is good medicine. PMID- 19301485 TI - Handbook of experimental pharmacology 192. Nicotine psychopharmacology. Preface. PMID- 19301487 TI - Dieting on a budget: plus the secrets of thin people, based on our survey of 21,000 readers. PMID- 19301488 TI - Spontaneous re-ossification of a large calvarial defect in an older child. PMID- 19301489 TI - [Modern approach to treatment of trophic ulcers of legs with perifocal varicose eczema, associated with mycotic infection]. AB - During 2005-2007 128 patients at the age from 21 to 80 years with extensive trophic ulcers of legs were treated in the contaminated surgery department of A.V. Vishnevsky Surgery Institute and Korolenko Hospital. In 87 patients (69.9%) varicosity was the cause of venous insufficiency, in 41 patients--post-thrombotic disease. 23 patients (17.97%) had double-sided throphic ulcers. In 50 patients (39.1%) ulcer area exceeded 40 cm2. In microscopical analysis fungi were discovered in 87 patients (89.06%). In the remaining 15 patients diagnosis was verified culturally. System etiotropic antimycotic antibacterial treatment including hyposensitizative, anti-inflammatory, antihistamine drugs was carried out. 34 patients (26.57%) received broad spectrum antibiotic from terbinafine group (Terbizil) in dosage 250 mg once a day during 3 moths with concurrent use of topical antimycotics. 34 patients (26.56%) recieved pulse therapy with broad spectrum antibiotic Rumicoz 400 mg a day during 7 days in combination with topical antimycotics. Nizoral 400 mg a day during 30 days was prescribed to 26 patients (20.3%). 34 patients (26.56%) were treated only with topical antimycotics. After preoperative preparation and reduction of inflammatory process 64 patients had underwent excision of trophic ulcers whereupon the therapy was continued. In these patients regress of varicose eczema manifestation was achieved on the 10th day. In patients who hadn't received surgical cure regress of varicose eczema was signed on the 23-25th day after onset of treatment. Question of plastic closing of throphic ulcers and wounds after their excision was solved particularly. 14 patients needed recurring surgical management becose inflammatory process was continued. Comprehensive approach to therapy taking into account mycotic semination appears to be appropriate and effective. Proactive surgical tactics (excision of throphic ulcer) allows to quicken preoperative preparation for correction of venous blood flow. PMID- 19301490 TI - [Antioxidants treatment of severe post-operative pyoinflammatory and septic complications]. AB - New medical human lactoferrin product called Laprot possessing antioxidant, detoxicant, anti-inflammatory immunomodulating properties was developed and registered (serial number LS-002374) in the P.A. Hertsen Institute. System (intravenous) administration of Laprot is efficacious detoxicant and anti inflammatory treatment in patients with severe postoperative pyoinflammatory and septic complications accompanied by polyorgan failure. Local administration contributes to clinically apparent cleansing of festering wounds and cavities, regress of local pyoinflammatory processes, reduction of local purulo-necrotic processes of trachea's mucosa. Laprot administrated intravenously as in the case of topical administration is well tolerated by the patients and doesn't cause any side affects. PMID- 19301491 TI - [Gall bladder drainage at acute pancreatitis]. AB - Determination of indications and contra-indications to cholecystostomy in patients with acute necrotic pancreatitis was the aim of this study. 102 patients were examined. Presence of caput pancreatis enlargement, infiltration and limited fluid accumulation, gall bladder enlargement and increase in bilirubin level was evaluated. Evidence of biliary hypertension was evaluated according to bile amount flowing out of the cholecystostoma. Findings allowed to draw the following conclusions. Increase in conjugated bilirubin, dilatation of bile ducts, gall bladder enlargement over 120 mm lengthwise are the indications to cholecystostoma surgery at acute necrotic pancreatitis. Gall bladder drainage in other cases is not reasonable. Cholecystostoma surgery should be carried out at the latest 3 days from the moment of admission to hospital preferably by laparoscopic method. Presence of initial hypotension is the contra-indication to gall bladder drainage even in case of total normalization of hemodynamics. PMID- 19301492 TI - [Laparascopic cholecystectomy in patients with acute cholecystitis]. AB - Experience of diagnostics and treatment of 758 patients with acute cholecystitis was summarized. Authors attach the main importance to evaluation of ultrasound data and functional condition of respiratory and cardio-vascular sistem choosing the method of surgical treatment. Sparse use of laparoscopic cholecystectomy in treatment of acute cholecystitis compared with chronic may be explouned by presence of complications in patients which make problems in differentiation of tubular structures during the operation. Authors offered to use ultrasound dissection for clear identification of tubular structures and argon coagulation for hemostasis and safe mobilization of gall bladder. Stick to suggested tactics authors practically doubled the amount of performed laparoscopic cholecystectomies reducing at the same time the frequency of complications. PMID- 19301493 TI - [Diagnostics and surgical tactics for Mirizzi syndrome]. AB - Analysis of clinical investigation and surgical treatment of 284 patients with Mirizzi syndrome (MS) was carried out. According to A.Csendes and co-authors classification (1989) 223 patients had the I type of MS, 49 patients--the II type, 9 patients--the III type, 3 patients--the IV type. Diagnosis was ascertained before the operation in 27.4% of patients; with the help of ultrasound study--in 9.1%, by means of retrograde cholangiopancreatography--in 18.3%. In other cases diagnosis was determined during the operation. Mode of the operation depended on the type of MS. Perioperative duct damage was signed in 4 cases (1.4%). Laparoscopic cholecystectomy (LCE) was carried out to 56 patients; cholecystectomy from mini-access--to 41 patients. Remaining 187 patients underwent abdominal operation. In 2 cases operation was finished by external duct drainage; in 7 Roo choledochojejunostoma was formed. Authors suggest that the I type of MS is not a contra-indication for LCE. In case of difficulties in LCE switch to cholecystectomy from mini-access is preferable. PMID- 19301494 TI - [Therapeutic approach to non-complicated non-parasitic hepatic cysts]. AB - Results of treatment of 378 patients with non-complicated non-parasitic hepatic cysts during the last 15 years are presented. Diagnostics was based on the developed algorithm of diagnostics and treatment of space-occupying lesions of liver. Therapeutic approach to non-parasitic hepatic cysts was defined on the basis of their sizes. At case of small cysts monitoring was carried out. During the period of supervision the authors didn't observe any complications. Increasing of the cyst sices was signed only in 7.94% of cases. Puncture and sclerotherapy under ultrasound control were carried out in medium cysts with 96.70% of efficacy. If the authors didn't deny the parasitic or tumorous nature of liver lesions carry out the surgical treatment, which was also considered to be indicated at case of sizcable enlargement of the non-parasitic hepatic cysts and inefficiency of mini-invasive treatment. PMID- 19301495 TI - [Possibilities of early diagnostics of echinococcosis relapse]. AB - Successful introduction of modern mini-invasive technologies of echinococcosis treatment necessitate problem-solving in early exposure of relapse and diagnosis verification at small cysts. It is ascertained that development rate of recurrent cyst decreases with size enlargement. Median of growth rate had amounted 1.5 mm a month (quartile--1.02 and 2.4 mm/month) till the diameter of the cyst reached 5 cm. Carrying out immune-enzyme analysis allows to forecast treatment results in terms of 3-3.5 years in most observations. Taking this data into account algorithm of early relapse diagnosis was worked out. Long-term results of surgical treatment of echinococcosis of various localization were studied in 652 patients. Regular check-up according to developed algorithm allowed to reveal relapses after 12.4 +/- 7.7 months on average, whereas in the absence of regular check up relapses were diagnosed after 27.9 +/- 15.6 months on average (p=0.0005). Revealed cysts were relatively bigger (3.8 +/- 1.1 and 8.0 +/- 3.7 cm respectively), that at regular check-up according to algorithm. PMID- 19301496 TI - [Hardware controlled laparostomy in staged treatment of peritonitis with abdominal compression syndrome]. AB - Clinical results of treatment were analyzed in 97 with patients generalized peritonitis. Application of staging hardware controlled laparostomy (HCL) allowed to halve the frequency of wound complication in comparison with semienclosed treatment mode. High clinical efficacy of control and correction of biophysical parameters of abdominal cavity was revealed: lethality from generalized peritonitis decreases in 2.4 times (from 39.3% to 14.3%) as a result of HCL application with monitoring and correction of intra-abdominal pressure in comparison with uncontrolled traditional treatment modes. Construction of the apparatus used for controlled laparostomy allows to open and close abdominal cavity under control of intra-abdominal pressure and tighting force of paravulnar tissues. PMID- 19301497 TI - [Fournier disease in surgeon's practice]. AB - Results of treatment of fulminating scrotum gangrene (Fournier disease) in 17 patients at the age of 34-82 years are presented. Bounded (slow progressive) forms were observed in 12 patients, widespread (fulminant and rapidly progressive) forms--in 5 patients. Colorectal pathology as the most likely cause of Fournier disease was observed in 8 patients, urogenital pathology--in 7 patients. In 2 patients blunt injury (1) and gunshot wound (1) of scrotum preceded the development of Fournier disease. 5 patients suffered from diabetes mellitus differed in severity. Non-clostridial anaerobic bacteria appeared to be the main infection agents causing purulo-necrotic lesion (88.2%). Lethality amounted 29.4% (5 patients from 17 died). Authors consider that active tactics applying on the aim of surgical management in complex with early and staged necrectomy and complex system of local management of the wound, used against a background of intensive multicomponent medical correction of homeostasis disturbances, allows to improve treatment results. PMID- 19301498 TI - [Parenteral nutrition in children with traumatic shock]. AB - Results of inclusion of parenteral nutrition in the complex of medical measures in 300 children at the age of 1-15 years with multitrauma complicated by shock are presented. Concepts of parenteral nutrition in children with traumatic shock are formulated. Indications, terms of parenteral nutrition onset are substantiated. Characteristics, dosages, rules of parenteral nutrition components apply in children with traumatic shock are described. Influence of developed regimen of parenteral nutrition on the course of traumatic shock on children is evaluated. PMID- 19301499 TI - [Forming of straight urointestinal anastomosis at enteric replacement of urinary bladder]. AB - Problem of decreasing the complications rate of intestinal urine derivation is very relevant. One of the most important problem is the stricture of urointestinal anastomosis. The problem of the choice of antireflux or direct method of ureters implantanion is not solved. Results of forming of 186 renoureteral units (RUU) according to direct method and 70 RUU according to antireflux method were analyzed by the authors. Straight anastomosis was applied according to Wallace 1 and 2 technique, antireflux--according to Abol-Enein technique or by means of transverse duplicating. Age of the patients ranged from 16 to 76 years (56 years on average); men prevailed in groups. Terms of supervision ranged from 6 to 108 months (54.6 months on average). Strictures and failure of ureteroreservoir anastomosis using Wallace method were not revealed (0%), then using antireflux methods (transverse duplicating or extramural Abol Enein tunnel) the frequency of complication amounted 2.8% on average. Strictures after antireflux mode in case of their early exposure were successfully corrected using endoscopic methods. In other cases open revision of anastomoses with their repeated apply was carried out. Reno-reservoir reflux was not observed in any cases after antireflux methods. Clinical manifestation of reflux was revealed at 2 RUU. Basing on the data renal radionuclide of image renal function didn't significantly change after direct or antireflux methods of ureters implantation. Wallace method is remarkable for its technical usability. This method ensures low level of complications after as incontinent and continent urine derivation. PMID- 19301500 TI - Advancing reproductive health of young people in the European region. AB - Evidence that neglect of the sexual and reproductive health needs and problems of young people can seriously jeopardize their health and future well-being is growing. Addressing the sexual and reproductive health needs and problems of adolescents is a crucial element of both WHO European Regional Strategy on Sexual and Reproductive Health and WHO Global Reproductive Health Strategy. Although the range of reproductive health challenges in young people includes a broad list of issues, this paper focuses on the key public health concerns that have a major impact on young people's future opportunities, such as adolescent pregnancy, HIV/AIDS and other sexually transmitted infections and on policy implications. As those challenges are largely related to unsafe sex behaviours, the paper is also looking at issues related to sexuality education, its effectivenes and policy development implications. PMID- 19301501 TI - [Demographic, social and health dimension of adolescent childbearing]. AB - Despite noticeable changes in the preferences concerning forming families and growing aspirations of young people with respect to level of education, professional and social status, thousands of adolescent pregnancies and births are still recorded, also in highly developed countries. Childbearing by adolescents--many of whom demonstrate insufficient social, psychological and physical maturity to handle the role of mothers--may involve numerous negative health and social consequences to the mother and her child, both immediate and in later life. The purpose of the article is to discuss the current situation with respect to teenage fertility in Poland and Western countries, and to present possible consequences of adolescent childbearing in demographic, health and social dimensions. The paper uses data taken from birth registers and data bases concerning review of literature covering the discussed issues, in particular from results of panel and retrospective studies enabling evaluation of the risk of specific consequences to the mother and her child. PMID- 19301502 TI - [Risk factors of early sexual initiation]. AB - PURPOSE: to present the relationship between early sexual initiation and factors related to family, school, peers, perceived health, type A behaviour and psychoactive substances use among 16-year-olds. MATERIAL AND METHODS: representative sample of students from first grade of upper secondary school (N = 1514; mean age 16.7 years) was randomly selected. In anonymous questionnaire questions from the international HBSC (Health Behaviour in School-aged Children) questionnaire and TAB scale for Type A behaviour assessment were used. Multivariate logistic regression models were estimated to assess the risk of early sexual initiation in relation to analysed factors. RESULTS: among 16-year olds 34.9% of boys and 20.6% of girls reported sexual initiation. In this group 37.3% of boys and 12.9% of girls had two or more sexual partners. Many risk factors of early sexual initiation were identified. The highest risk was in boys in the case of frequent drinking of vodka and beer, using amphetamine, truancy and among pupils from basic vocational schools. Among girls the highest risk was connected with frequent truancy, daily smokers, and those who spent a lot of time out of home. Many risk factors were related to peers and psychoactive substance using. CONCLUSIONS: adolescents who started their sexual life are a group of high risk of sexual and reproductive health problems and they need medical care, including psychological counselling. The knowledge about risk factors of early sexual debut and monitoring trends in their change should be taken into consideration in planning prevention programmes and school sexual education. PMID- 19301503 TI - [Socio-economic determinants of physical development disorders among 15-year-olds in Poland]. AB - AIM of the study was to examine the association between family socio-economic status, local area problems and prevalence of underweight, overweight and obesity in Polish 15-year-olds. MATERIAL AND METHODS: data from HBSC (Health Behaviour in School-aged Children) surveys, carried out in Poland in 2006 in representative sample of adolescents mean aged 15.7 (N = 2287) was used. An international standard questionnaire has been chosen as an instrument for data collection. Three indicators of socio-economic status (SES) were developed by researchers in HBSC study: FAS (Family Affluence Scale), PFW (Perceived Family Wealth) and LAP (Local Area Problems). Prevalence of underweight, overweight and obesity was estimated on the base of BMI national standards and WHO criteria. Comparative analyses were done by chi2 test and multivariate logistic regression models were used. RESULTS: the prevalence of overweight and obesity was higher in boys (12.9%) than in girls (8.7%, p < 0.001) and almost in the same frequency in urban and rural areas. Underweight was indicated in 4.4% 15-year-olds, more frequently in girls and in urban adolescents. Most of boys with overweight and obesity were from poor families by using FAS, especially in rural area (15%), and girls from poor families by using PFW (15.5%, p = 0.05). Gender and LAP were the strongest predictors of overweight and obesity. Underweight occurred more frequently in adolescents from rich families than from poor ones (for girls p = 0.023), in urban inhabitants and in the areas, where more local problems were observed. CONCLUSIONS: Site of domicile is the differentiation's factor for underweight only, masculine gender is the independent predictor for overweight and obesity and status of local area is the only factor which has an effect on both underweight and overweight and obesity occurrence PMID- 19301504 TI - [Subjective health of adolescents aged 11-15 years in Poland and other countries of the European Union]. AB - THE AIM: to present results of the study concerning subjective health of adolescents aged 11, 13 and 15 years in 26 countries members of the European Union (EU) in 2005/06 and changes in perception of health among Polish adolescents between 2002 and 2006. METHOD: data from the 2005/06 HBSC survey (Health Behaviour in School-aged Children: A WHO Collaborative Cross-national Study), carried out in 26 EU countries among 11-, 13- and 15-year-olds (N = 140,339) were analysed. For Polish adolescents results of two surveys from 2002 and 2006 were compared. The international standard questionnaire was used. Three subjective indicators of health were used: self-rated health, life satisfaction (using 0-10 points Cantril scale) and 8 subjective health complaints (headache, stomach-ache, back-ache, feeling low, irritability or bad temper, feeling nervous, difficulties in getting to sleep, feeling dizzy). RESULTS: subjective indicators of health markedly differ between adolescents in different EU countries. In all countries adolescents report recurrent (every day or more then once a week) somatic and psychological complaints. Multiple complaints (3 or more) experienced 8% of boys and 38% of girls. There are strong gender differences in subjective health. In all countries girls perceived their health worse than boys. In Polish adolescents some tendency of improvement of subjective health indicators was observed between 2002-2006. CONCLUSION: Existing differences in adolescents' subjective health in EU countries and gender differences are probably determined by many cultural, social and economical factors. These differences create health inequalities and probably will be maintained for a long time. PMID- 19301505 TI - [Social determinants of psychological distress in adolescents aged 18 years in Poland]. AB - THE PURPOSE of the study was to analyse the relationships between social environment (school, family, peers) factors and distress of adolescents aged 18 years. MATERIAL AND METHODS: the questionnaire survey was carried out in the sample of 1379 pupils in the third grade of upper secondary schools; mean age of pupils 18.7 years. The questionnaire contained scale GHQ-12 (General Health Questionnaire) and questions from HBSC (Health Behaviour in School-aged Children. A WHO Collaborative Cross-national Study) survey, concerning school attitude, family structure, parental bonding, communication with parents and peers' relationships. Comparative analyses were done by chi2 test. Odds ratio and multivariate logistic regression models were estimated for risk assessment. RESULTS: over 40% of 18 year olds had a higher level of psychological distress, significantly more frequent among girls than boys (p < 0.001), and urban than rural adolescents (p < 0.01). Pupils from general upper secondary schools more often felt distress than pupils from basic vocational schools (50% vs. 27%, p < 0.001). Girls who reported not liking school and felt pressured by schoolwork had a higher level of distress (Exp(B) = 1.8 and 2.1). Significant relationships were found between worse parental bonding, difficulties in communication with parents and higher level of distress both in boys and girls (Exp(B) from 1.8 to 2.8). Lack of same-sex close friends and no time spent with them after school were predictors of higher distress mostly in boys (Exp(B) od2,2 do 11.5). CONCLUSION: school problems (prospect of school-leaving e xams), difficult family relationships and especially lack of same-sex friends and spending time with them outside school are risk factors for psychological distress of youth aged 18 years. PMID- 19301506 TI - [Positive health behaviours scale for adolescents: initial psychometric analysis]. AB - PURPOSE: to elaborate the Positive Health Behaviours Scale (PHBS) for use with adolescents and to provide its psychometric analysis. METHODS: the instrument consists of 30 statements comprised of five subscales (nutrition, relaxation and behaviours connected with psychosocial health, safety, taking care of one's body and physical activity) which were measured on a 4-point summated rating scale from 3 (almost always) to 0 (almost never). Psychometric analysis included assessment of reliability (using Cronbach's a coefficient Joreskog's rho coefficients), theoretical validity (factor analysis) and construct validity (using data concerning perceived health, self-assessment of care of own health, psychoactive substance use and subjective health complaints; four hypothesis were tested) and unidimensionality (principal components). The sample consisted of 307 adolescents aged 13-17. RESULTS: Cronbach's alpha coeffient for the whole scale amounted to 0.823 but for three out of five subscales it was lower than the accepted value of 0.6. Joreskog's rho coefficient was at satisfactory level for all subscales. Factor structure of the scale was different from what was expected. Construct validity of the subscales was satisfactory--all four hypotheses were confirmed. All subscales were also unidimensional. CONCLUSIONS: psychometric analysis of PHBS for adolescents did not give univocal results concerning its quality. Only reliability of the whole scale was satisfactory. Further study with application of larger, randomly selected sample of adolescents is required. The scale can be used as a tool in health education and in prophylactic health studies of adolescents. PMID- 19301507 TI - [Theory of resilience. Key conceptual constructs and chosen issues]. AB - The aim of this article is to familiarize readers with key conceptual constructs related to resilience theory. This theory explains the phenomenon of children's and adolescents' positive adaptation despite various adverse life conditions and traumatic events. The resilience theory emphasizes the importance (relevance) of positive factors and mechanisms for child and adolescent development. Thus, it could be very useful for developing primary prevention and mental health promotion programmes among children and youth. This article is based on a review of publications written by significant resiliency researchers: M Rutter, N. Garmezy, E. Werner, S. Luthar, A. Sameroff, K. Kumpfer, A. Masten, M. Zimmerman, D. Cicchetti. More than 20 articles and book chapters published during the past 25 years were taken into consideration. They were chosen from the electronic database available at the University of Michigan and SAMSHA, and publications available in Poland. Several resilience definitions are mentioned, indicating the interactive and dynamic process of positive adaptation. These definitions encompass the impact of both risk and positive factors (and their interaction) on the individual's behavior, competence and health. The key conceptual constructs such as risk, risk factors, positive factors, resilience models and mechanisms are described in this article. The differences between the risk and protective mechanism are also presented. The difficulties related to operationalization of the key resilience constructs in empirical research are discussed. Common difficulties are related for example to clear criteria for risk and positive adaptation, and for risk group selection. PMID- 19301508 TI - [Resilience--from theory to empirical research]. AB - Resilience refers to mechanisms that enable young people to sustain good health and harmonic development despite adversities. THE AIM: to present resilience theory and one example of empirical evidence. MATERIAL AND METHODS: the survey was carried out in 2006, on the representative sample of 2287 adolescents (average age 15.7) in the frame of HBSC (Health Behaviour in School-aged Children) study. As an outcome measure, a syndrome of two or more subjective complaints experienced more than once a week was selected. We assumed that negative perception of peer environment could be a risk factor. A scale measuring use of psychoactive substances, categorized into 2 levels, was applied. As protective factors, two dichotomous variables describing family were chosen: easiness to talk to parents and quality of family relationships. RESULTS showed that 29.2% boys and 45.9% girls suffered from multiple subjective complaints. The prevalence of these complaints increased in adolescents representing negative perception of peer environment, compared to the most positive perception (for girls: from 37.9% to 57.5%, p < 0.001; for boys: from 19.7% to 39.1%, p < 0.001), but the rate of change was lower in more supportive families. Logistic regression model indicated significant interaction between risk and protective factors, but was valid only in boys and in the model including quality of family relationships. CONCLUSIONS: Good family relationships are recognized as a protective factor reducing negative influence of peers on adolescent subjective health. It is important to reach the group of adolescents reporting accumulation of poor family relationships and a high risk peer environment. PMID- 19301509 TI - [Menstrual irregularities in puberty]. AB - The establishment of regular menstrual cycle is an important process for an adolescent girl. Psychologically it is a sign of becoming an adult and a factor for female identity. Normal function of the hypothalamic-pituitary-ovarian axis and system of sexual hormones is observed, which are the basis for future fertility and reproduction. Disturbances of this regulatory system during its development, cause dysfunctional uterine bleeding and are a frequent gynaecological problem. The challenge is to distinguish normal individual variation from real endocrine or organic problems. In the article the most common menstrual disorders with the focus on dysfunctional uterine bleeding in puberty, its recognition and treatment are presented. PMID- 19301510 TI - [Chronic pelvic pain in adolescent girls]. AB - Chronic pelvic pain is a serious problem in paediatric and adolescent gynaecology, because it reaches approximately 10-20% level among outpatients. In diagnostics of chronic pelvic pain in adolescent girls both gynaecological and non-gynaecological reasons should be considered. The diagnostic process should include: a medical history, physical examination, gynaecological examination (rectal or vaginal examination), laboratory tests and imaging studies (ultrasonography, magnetic resonance). Moreover, in the diagnostic process a laparoscopy is recommended to be carried out in girls and teenagers with chronic pelvic pain. The therapy of chronic pelvic pain in adolescent girls involves: pharmacotherapy, surgical methods (laparoscopy/laparotomy), alternative pain therapy and psychotherapy. PMID- 19301511 TI - Recommendations of the Polish Society of HPV Infections Prophylaxis - (PS-HPVIP) (Polskie towarzystwo profilaktyki zakazen HPV - PTPZ-HPV) concerning the use of preventive vaccines against HPV. PMID- 19301512 TI - The efficacy of a new dentifrice with caries, plaque, gingivitis, calculus, tooth whitening, and oral malodor benefits. PMID- 19301513 TI - Clinical investigation of the antiplaque efficacy of a new variant of a commercially available triclosan/copolymer/fluoride dentifrice. AB - OBJECTIVE: The objective of two single-blind, three-treatment, crossover design, clinical studies was to evaluate the antiplaque efficacy using the Modified Gingival Margin Plaque Index (MGMPI) scores of three dentifrices: 1) a dentifrice containing 0.3% triclosan/2.0% polyvinylmethyl ether/maleic acid (PVM/MA) copolymer/sodium fluoride in a 17% dual silica base (Colgate Total Advanced Toothpaste-Test Dentifrice); 2) a commercially available dentifrice containing 0.3% triclosan/2.0% PVM/MA copolymer/sodium fluoride in a 10% high-cleaning silica base (Colgate Total Toothpaste-Positive Control Dentifrice); and 3) a commercially available dentifrice containing 0.243% sodium fluoride in a regular silica base (Colgate Winterfresh Gel-Negative Control Dentifrice). METHODS: In each study, subjects reported to the clinical facility, and those who met the inclusion/exclusion criteria were given a complete oral prophylaxis, a soft bristled toothbrush, and a commercially available dentifrice (Colgate Cavity Protection Fluoride Toothpaste). They were instructed to use these products exclusively for seven days (washout period), after which time they reported back to the clinical facility and were randomized into three treatment groups. All subjects then brushed their teeth for one minute with a full ribbon (approximately 1.5 gm) of Colgate Cavity Protection Fluoride Toothpaste, and immediately followed with a one-minute brushing using a full ribbon of one of the three study dentifrices. Subjects then rinsed with a red disclosing solution (Butler Red-Cote) and had their teeth and gums examined to assess their plaque content. They returned to the clinical facility after 24 hours of no oral hygiene to again have their teeth and gums examined to assess their plaque content. As per the crossover clinical design, the same methods and materials were used until all subjects used all three study treatments. RESULTS: Seventeen subjects in the first study and 16 subjects in the second study complied with the protocol and completed all phases of the study. Two-way ANOVA results from both studies showed that there was no difference in mean delta MGMPI scores between the groups using the Test Dentifrice and the Positive Control Dentifrice. Results also showed that there was a statistically significant difference (p < 0.05) in delta MGMPI scores between both the Test Dentifrice treatment and the Positive Control Dentifrice treatment when compared to the Negative Control Dentifrice. CONCLUSION: A new improved dentifrice containing 0.2% triclosan/3.0% PVM/MA copolymer/sodium fluoride in a 17% dual silica base is comparable in controlling dental plaque when compared to a Positive Control Dentifrice containing 0.3% triclosan/2.0% PVM/MA copolymer/sodium fluoride in a 10% high-cleaning silica base, and is statistically significantly better in controlling dental plaque when compared to a Negative Control Dentifrice containing 0.243% sodium fluoride in a silica base. PMID- 19301514 TI - A clinical investigation of the efficacy of two dentifrices for controlling established supragingival plaque and gingivitis. AB - OBJECTIVE: To assess the efficacy of a dentifrice containing 0.3% triclosan/2.0% polyvinylmethyl ether/maleic acid (PVM/MA) copolymer/0.243% sodium fluoride in a 17% dual silica base (Colgate Total Advanced Toothpaste-Test Dentifrice) for controlling established supragingival plaque and gingivitis, relative to that of a commercially available dentifrice containing 0.243% sodium fluoride in a silica base (Crest Cavity Protection Toothpaste-Control Dentifrice). METHODS: Following a baseline examination for supragingival plaque and gingivitis, qualifying adult male and female subjects from the Barcelona, Spain area were randomized into two treatment groups. Subjects were given a complete oral prophylaxis and instructed to brush their teeth twice daily (morning and evening) for one minute with their assigned dentifrice and a soft-bristled toothbrush. Examinations for supragingival plaque and gingivitis were repeated after three months and six months of product use. RESULTS: Ninety-four (94) subjects complied with the protocol and completed the study. Relative to the Control Dentifrice group, the Test Dentifrice group presented statistically significantly lower whole-mouth plaque index and gingival index scores (18.6% and 15.8%, respectively) after three months of product use. Similarly, relative to the Control Dentifrice group, the Test Dentifrice group exhibited statistically significant reductions in plaque index and gingival index scores measured on proximal sites (17.0% and 16.3%,respectively), and statistically significant reductions in plaque severity index and gingivitis severity index scores (22.2% and 46.9%, respectively) after three months of product use. Relative to the Control Dentifrice group, the Test Dentifrice group presented statistically significantly lower whole-mouth plaque index and gingival index scores (23.4% and 21.3%, respectively) after six months of product use. Similarly, relative to the Control Dentifrice group, the Test Dentifrice group exhibited statistically significant reductions in plaque index and gingival index scores measured on proximal sites (21.1% and 23.0%, respectively), and statistically significant reductions in plaque severity index and gingival severity index scores (27.1% and 64.5%, respectively) after six months of product use. CONCLUSION: The overall results of this double-blind clinical study, conducted in accordance with the quantitative criteria established by the American Dental Association, support the conclusion that a dentifrice containing 0.3% triclosan/2.0% PVM/MA copolymer/0.243% sodium fluoride in a 17% dual silica base is efficacious for the control of established supragingival plaque and gingivitis. PMID- 19301515 TI - Clinical studies to assess the extrinsic stain prevention and stain removal efficacy of a variant of a commercially available dentifrice containing a new dual silica system. AB - OBJECTIVE: The objective of these two six-week, double-blind clinical studies was to compare the extrinsic stain prevention (Study #1) and stain removal (Study #2) efficacy of a new dentifrice (Colgate Total Advanced Toothpaste) containing 0.3% triclosan/ 2.0% polyvinylmethyl ether/maleic acid (PVM/MA) copolymer/0.243% sodium fluoride in a 17% dual silica base, to a commercially available fluoride dentifrice without tooth whitening ingredients (Crest Cavity Protection Toothpaste) containing 0.243% sodium fluoride in a silica base. METHODS: Following baseline examinations for extrinsic tooth stain and an oral tissue examination, qualifying adult male and female subjects from the northern and central New Jersey, USA areas were randomized for each study into two treatment groups which were balanced for gender and level of extrinsic tooth stain. Subjects received a full oral prophylaxis at the start of Study #1 only. All subjects were provided with their assigned product and a soft-bristled adult toothbrush for home use. Subjects were instructed to brush their teeth for one minute twice daily (morning and evening) using only the dentifrice and toothbrush provided, and to refrain from using any other oral hygiene products for the entire six weeks of the study. There were no restrictions regarding diet or smoking habits during the course of the study. Examinations for extrinsic tooth stain and oral tissue assessments were repeated after three weeks and six weeks of product use. RESULTS: One-hundred fourteen (114) subjects participating in Study #1 and 119 subjects participating in Study #2 complied with the protocol and completed the entire study. In both studies, at the three-week and the six week examinations, subjects who used the new dentifrice exhibited statistically significantly lower levels of extrinsic tooth stain area and extrinsic tooth stain intensity than did those subjects who used the commercially available fluoride control dentifrice. CONCLUSION: A new dentifrice containing 0.3% triclosan/2.0% PVM/MA copolymer/0.243% sodium fluoride in a 17% dual silica base is more efficacious in the prevention and removal of extrinsic tooth stain than a commercially marketed fluoride dentifrice. PMID- 19301516 TI - A clinical investigation of the efficacy of two dentifrices for the reduction of supragingival calculus formation. AB - OBJECTIVE: The objective of this double-blind clinical study, conducted in harmony with Volpe-Manhold design for studies of dental calculus, was to compare the efficacy of a dentifrice containing 0.3% triclosan/2.0% polyvinylmethyl ether/maleic acid (PVM/MA) copolymer/0.243% sodium fluoride in a 17% dual silica base (Colgate Total Advanced Toothpaste) to that of a commercially available dentifrice containing 0.243% sodium fluoride in a silica base (Crest Cavity Protection Toothpaste) with respect to the reduction of supragingival calculus formation. METHODS: Adult male and female subjects from the San Francisco area were entered into the eight-week pre-test phase of the study. Subjects received an evaluation of oral soft and hard tissues and were given a complete oral prophylaxis. They were provided with a non-tartar control placebo dentifrice and a soft-bristled adult toothbrush, and were instructed to brush their teeth twice daily (morning and evening) for one minute. After eight weeks of using the placebo dentifrice, subjects were examined for baseline supragingival calculus formation using the Volpe-Manhold Calculus Index. Qualifying subjects were randomized into two treatment groups which were balanced for gender and baseline calculus scores. All subjects entered into the twelve-week test phase were given a complete oral prophylaxis, and were provided with their assigned dentifrice and a soft-bristled adult toothbrush for home use. Subjects were instructed to brush their teeth for one minute twice daily (in the morning and evening). Prior to each study visit, subjects refrained from brushing their teeth and eating and drinking for four hours. RESULTS: Seventy-seven (77) subjects complied with the protocol and completed the study. At the twelve-week examination, the Test Dentifrice group presented a mean Volpe-Manhold Calculus Index score of 13.22 and the Control Dentifrice group presented a score of 20.29. After twelve weeks of product use, the Test Dentifrice group exhibited 34.8% less supragingival calculus formation than the Control Dentifrice group (statistically significant at p < 0.05). CONCLUSION: The overall results of this double-blind clinical study support the conclusion that after twelve weeks' use of a dentifrice containing 0.3% triclosan/2.0% PVM/MA copolymer/0.243% sodium fluoride in a 17% dual silica base provides significantly greater control of supragingival calculus formation relative to that of a commercially available dentifrice containing 0.243% sodium fluoride in a silica base. PMID- 19301517 TI - A clinical investigation of the efficacy of two dentifrices for controlling oral malodor and plaque microflora overnight. AB - OBJECTIVE: The objective of this independent, double-blind, overnight clinical study was to assess the efficacy of a dentifrice containing 0.3% triclosan/2.0% polyvinylmethyl ether/maleic acid (PVM/MA) copolymer/0.243% sodium fluoride in a 17% dual silica base (Colgate Total Advanced Toothpaste) for the overnight control of oral malodor and plaque microflora, relative to that of a commercially available dentifrice containing 0.243% sodium fluoride in a silica base (Crest Cavity Protection Toothpaste). METHODS: After a five-day period using a supplied commercially available fluoride toothpaste, adult male and female subjects from the Chengdu, China area reported to the clinical facility in the evening, after having refrained from the use of all oral hygiene procedures, as well as eating and drinking, for at least six hours. Following an assessment of the oral soft and hard tissues, subjects were evaluated for baseline oral malodor by a panel of four trained judges using a nine-point hedonic scale. Plaque samples (baseline) from the teeth on the left side of subjects' mouths were also collected. Subjects who presented a mean baseline oral malodor score between 6.0 and 8.4 were stratified according to their oral malodor score and randomly assigned within strata to one of the two treatment groups. Subjects were provided with their assigned dentifrice and instructed to brush their teeth on the left side of their mouth only, for one minute under supervision. Subjects returned to the testing facility the following morning for an overnight evaluation, having refrained from the use of all oral hygiene procedures, and from eating and drinking. An oral malodor assessment was performed, plaque samples were collected from the teeth on the right side of the mouth, and a final oral soft and hard tissue evaluation was performed. RESULTS: Eighty-one (81) subjects completed the study. At the overnight evaluation, subjects in the triclosan/copolymer/fluoride dentifrice group and the fluoride dentifrice group presented mean oral malodor scores of 4.91 and 6.86, respectively. Relative to the fluoride dentifrice group, at the overnight evaluation the triclosan/copolymer/fluoride dentifrice group presented a statistically significant 28.4% reduction in oral malodor scores. Additionally, at the overnight evaluation, subjects in the triclosan/ copolymer/fluoride dentifrice group and the fluoride dentifrice group presented geometric mean microbial colony forming unit (CFU) scores of 3.15 and 6.07, respectively. Relative to the fluoride dentifrice group, at the overnight evaluation the triclosan/copolymer/ fluoride dentifrice group presented a statistically significant 49.5% reduction in microbial CFU scores. CONCLUSION: The overall results of this double-blind clinical study support the conclusion that a single evening's use of a dentifrice containing 0.3% triclosan/2.0% PVM/MA copolymer/0.243% sodium fluoride in a 17% dual silica base provides statistically significantly lower levels of overnight (12-hour) oral malodor and plaque microflora when compared to a commercially available dentifrice containing 0.243% sodium fluoride in a silica base. PMID- 19301518 TI - Patient care recognised. PMID- 19301519 TI - Logic, behaviour and dentistry. Interview by Paul O'Grady. PMID- 19301520 TI - Factors to consider in the transition to digital radiological imaging. AB - The dentist considering adopting digital radiological technology should consider more than the type of detector with which to capture the image. He/she should also consider the mode of display, image enhancement, radiation dose reduction, how the image can be stored long term, and infection control. PMID- 19301521 TI - Supernumerary teeth among Irish school children attending the public orthodontic service in Cork and Kerry. PMID- 19301522 TI - Results of a peer review process: the distribution of codes by examining dentists in the Republic of Ireland 2006-2007. AB - The Health Service Executive (HSE) appointed 20 examining dentists in April 2006 under contract for one year as part of a probity assurance initiative by peer review in the Dental Treatment Services Scheme (DTSS) in the Republic of Ireland. AIM: The aim of the study was to analyse the distribution of codes assigned to the reports drawn up by the examining dentists. METHODS: At the end of the year's contract, each examining dentist forwarded an end of contract report of their activity, including the distribution of codes issued, to the HSE. These were correlated into a national summary of examining dentist activity, from which the data used in the study was extracted. A total of 11 different codes were used, varying from an indication of agreement between the examining dentist and the contracting dentist (code A) to a significant disagreement (code D). RESULTS: The vast majority (94.5%) of reports on the clinical examination of patients, drawn up by examining dentists, were in broad agreement with the treatment or estimate of the responsible contracting dentists. A total of 622 contracting dentists received such reports. The small minority of reports (4.8%) where there was a significant disagreement related to a small number of dentists (47 dentists). CONCLUSIONS: The study provides evidence that most contracting dentists were not a probity risk. The author suggests that any future probity assurance initiative should focus on areas of high risk rather than random selection of patients/contracting dentists. An enhanced advisory role for the examining dentist is recommended. PMID- 19301523 TI - Let's get digital. PMID- 19301524 TI - Are you ready for that rainy day? AB - When taking out income protection, always insist that the company providing it will cover you for your 'own' occupation and not just 'any' occupation or a 'suited' one. Be aware of the deferred period or 'waiting time' and try and get some 'Day 1' in place. The younger a dentist decides to take out income protection, the better. Not only will they secure cover at a reasonable price now, but they will also be able to maintain it for their whole career. PMID- 19301525 TI - Caries management in the dental practice. AB - Practical, cost-effective implementation of caries management in general practice has been limited by several factors. The single pathogen model of disease has not been effective in clinical caries management, and the advent of the ecologic plaque model and a better understanding of the management of imbalances in dental biofilms have led to the development of more effective treatment protocols based on the elevation of oral pH. Simplification of caries risk assessment, in combination with more effective treatment regimens, means effective caries management can be readily provided by general dentists. These gains in efficiency and efficacy, in combination with applicable current dental terminology (CDT) codes, means that caries management has become economically viable in private practice. PMID- 19301526 TI - Dental plaque as a biofilm: the significance of pH in health and caries. AB - Dental plaque is an example of a biofilm; its presence is natural and it supports the host in its defense against invading microbes. In health, the microbial composition of dental plaque is diverse and remains relatively stable over time (microbial homeostasis). The predominant microorganisms prefer host molecules (eg, salivary mucins) and a neutral pH for growth. Under certain circumstances, this microbial homeostasis can break down and diseases such as caries can occur. In dental caries, there is a shift toward increased proportions of acid-producing and acid-tolerating species, such as mutans streptococci and Lactobacilli, although other species with relevant traits can participate in demineralization. Strategies to control caries include effective oral hygiene practices to reduce biofilm development, and adoption of a low-sugar diet to restrict periods of acidic challenge to teeth. These conventional approaches also should be augmented by interference with the factors that enable the cariogenic bacteria to outcompete the organisms associated with health. Evidence suggests that regular conditions of low pH in plaque select for mutans streptococci and Lactobacilli. Therefore, the suppression of sugar catabolism and acid production by the use of metabolic inhibitors in oral care products, the consumption of nonfermentable sweeteners in snacks, the stimulation of saliva flow, and/or other strategies that maintain supragingival plaque at a pH around neutrality will assist in the maintenance of microbial homeostasis in plaque. PMID- 19301527 TI - A clinician's guide to CAMBRA: a simple approach. AB - Caries risk assessment identifies those factors that are pathologic and suggests ways an astute clinician can implement protective strategies that can prevent progression and/or return the patient to health. Caries management by risk assessment (CAMBRA) focuses on treating and preventing the cause of the disease at an early stage, rather than waiting until it causes damage to tooth structure. This article summarizes and simplifies information previously published about CAMBRA implementation from the perspective of today's practicing clinician. The most recent science on prevention, remineralization, antimicrobials, and pH, as well as the use of fluoride, xylitol, and casein phosphopeptide-amorphous calcium phosphate (CCP-ACP) is also discussed. PMID- 19301528 TI - Managing caries in the high-risk child. AB - Dentists often report that treating children in their practices is either the most rewarding or the most challenging of all their professional experiences. Special considerations must be made for the biology of the caries process, the psychological development of the child, parental involvement, the complex environmental factors that contribute to caries, and the wide variety of clinical treatment options available. Caries management by risk assessment (CAMBRA) is becoming the standard of care for dental professionals. Although significant progress has been made against caries in the past 30 years, very resistant pockets of disease remain in certain demographics. This article presents a number of different clinical scenarios, which will explore diagnostic and restorative protocols that apply to children who are at risk for caries. PMID- 19301529 TI - Methodological issues in studying an insular, traditional population: a women's health survey among Israeli haredi (ultra-Orthodox) Jews. AB - This article describes obstacles encountered and strategies devised in planning and conducting a national telephone health survey (n = 459) of an insular, deeply traditional religious population, haredi (ultra-Orthodox Jewish) Israeli women. The paper discusses how special characteristics of this population influenced study design, sampling, data collection, and interpretation. Sampling employed polling data to identify haredi concentrations. Despite haredim's reputation for low survey participation, we achieved a 71-74% response rate (depending on the unknown eligibility of 24 phones never answered) in interviews conducted in 2003 2004. We describe our systematic attention to special aspects of haredi culture such as: modesty and speech codes; the need for rabbinic endorsement; and the importance of female, haredi interviewers. This research was initiated and managed by a community-based women's health non-governmental organization, in partnership with trained researchers. Our experiences can guide others surveying insular communities, such as traditional Muslim and Christian societies. PMID- 19301530 TI - Are lesbians really women who have sex with women (WSW)? Methodological concerns in measuring sexual orientation in health research. AB - Varying measures of sexual orientation are used in women's health research. As they incorporate different dimensions, definitions, and categorical groupings, the comparability of results obtained across studies using different measures remains unknown. We examined the comparability of results using data from the U.S. 2002 National Survey of Family Growth (n = 6,356). Women were classified according to sexual orientation identity, sex of sex partners in the past year, and sex of sex partners over the lifetime. Associations with six health outcomes were compared across sexual orientation schemes. Associations differed in magnitude and statistical significance, even producing conflicting results. Our analyses resulted in a series of methodological recommendations for research on sexual minority women. Data on both behavioral and identity measures should be gathered in health research; identity groups should not be combined for analysis; and researchers should carefully consider which classification scheme(s) to use based on the theoretical basis for the study and the implications for informing interventions. PMID- 19301531 TI - Breast health behavior among lesbians: the role of health beliefs, heterosexism, and homophobia. AB - The Health Belief Model was used as a conceptual framework for exploring how health beliefs and perceptions of heterosexism and homophobia in health care related to lesbians' breast health behaviors, including use of breast exams and mammograms, visits to health care providers, and use of complementary/alternative care. A total of 173 exclusively homosexual women completed survey measures assessing health beliefs, experiences of heterosexism/homophobia, and health maintenance behaviors as these related to breast health. Findings indicated that women perceived heterosexism and homophobia from providers to influence the amount of discussion they had with providers and, to a lesser extent, care received, frequency of service use, and health outcomes. Perceptions of heterosexism and homophobia in health care settings significantly contributed, beyond routine health beliefs, to women's use of breast self-exams, visitation to health care providers, and use of complementary/alternative care. Implications for culturally competent care are discussed. PMID- 19301532 TI - The menstrual cycle and sexual behavior: relationship to eating, exercise, sleep, and health patterns. AB - Patterns of eating, exercise, sleep, and health were investigated across 180 menstrual cycles of 89 women who engaged in sex with a male (n = 45; cycles = 85), a female (n = 21; cycles = 37), or abstained from sex (n = 33; cycles = 58) from January 2005 to December 2007 (10 contributed to 2 groups). Cycles were divided into 5 phases based on their luteinizing hormone surges. Daily questionnaires and saliva for IgA and cortisol analyses were obtained. Women indicated that they ate more (p < .008) and did not sleep as well (p = .02) during their luteal and premenstrual phases. Participants were less likely to experience food cravings and did not satisfy their cravings when they were ovulatory (p < .001). Additionally, a greater proportion of lesbians skipped breakfast (p = .01) and exercised less than heterosexuals (p = .05). Sexually active women had lower cortisol and IgA levels than abstinent women (p = .02). Our study discovered, and confirmed, systematic differences in eating, sleeping, and health patterns across women's menstrual cycles. PMID- 19301533 TI - Sociodemographic, psychological and health-related factors associated with poor mental health in Spanish women and men in midlife. AB - BACKGROUND: The purpose of this work was to examine the sociodemographic, psychological, and health-related factors (considered jointly) associated with poor mental health in midlife and to analyze whether risk and protective factors differed in men and women. METHODS: A cross-sectional study was conducted with a middle-class sample of 252 women and 189 men between 45 and 65 years of age from Spanish rural areas. Mental health status was measured with the 12-Item General Health Questionnaire (GHQ-12; Goldberg & Williams, 1988). Multiple logistic regression models were used to calculate odds ratios with confidence intervals of 95%, adjusting for confounding variables. RESULTS: The percentage of women (14.3%) with poor mental health was twice that of the men (7.4%). In women, the following variables were significantly and positively related to poor mental health: consumption of psychoactive drugs, physiological and cognitive anxiety; self-esteem and family satisfaction were protective factors. For men, physical complaints and cognitive anxiety were significant risk factors, and job satisfaction was a protective factor. CONCLUSIONS: In general, the psychological variables were more clearly related to poor mental health. Women had a more unfavorable profile, and the variables related to poor mental health differed for men and women, perhaps due to social roles associated with gender. To facilitate diagnosis and take preventive measures, men's and women's risk factors for poor mental health should be differentiated. PMID- 19301534 TI - Psychiatric comorbidity in women with disordered eating behavior: a national study. AB - This study used data collected by Statistics Canada in the Mental Health and Well being cycle of the Canadian Community Health Survey (N = 20,211) to examine the associations between disordered eating behavior and selected mood, anxiety and substance use disorders in adult women grouped in three age groups. Significant associations between disordered eating behaviors and major depression, substance dependence and psychological distress were found for women in all three age ranges. Prevalence of specific classes of anxiety disorders in women who reported disordered eating behaviors differed according to their age. Having disordered eating behaviors was significantly associated with both 12-month and lifetime prevalence of social phobia for women above 25 years of age. Lifetime prevalence of agoraphobia was associated with disordered eating behaviors in women 15-44 years old, and the 12-month prevalence was associated with disordered eating behaviors only in women 15-24 years of age. Disordered eating behaviors were significantly associated with lifetime prevalence of panic disorder in women in all three age ranges and with 12-month prevalence of panic disorder in women in two age ranges, 15-24 and 45 and above. The findings highlight the importance of taking age into consideration when developing assessment instruments and treatment strategies to address the co-occurrence of eating disorders and other psychiatric disorders. PMID- 19301535 TI - Marriage amendment ballots of 2004: could state residents read them? AB - Close to 43% of adults have difficulties reading and understanding printed materials. Given the fact that so many American adults have difficulty reading basic text, we were interested in looking at the readability levels of the marriage amendment ballots of the 2004 election. Results indicated, with the exception of two states, a mismatch between the educational levels and the readability levels of the proposed amendments. Legal recognition of same-sex civil marriages can be considered a mental health public policy issue and therefore public health professionals should advocate for easier to read and understand ballots. PMID- 19301536 TI - Similarity or dissimilarity in the relations between human service organizations. AB - Exchange theory and homophily theory give rise to counteracting expectations for the interaction between human service organizations. Based on arguments of exchange theory, more interaction is expected between dissimilar organizations having complementary resources. Based on arguments of homophily theory, organizations having similar characteristics are expected to interact more. Interorganizational relations between human service organizations in two regional networks in Flanders are examined in this study. Results indicate that human service organizations tend to cooperate more with similar organizations as several homophily effects but not one effect of dissimilarity were found to be significant. The results of this study contribute to the understanding of interorganizational networks of human service organizations and have implications for the development of integrated care. PMID- 19301537 TI - Institutional trust as a determinant of anxiety during the SARS crisis in Hong Kong. AB - The SARS crisis of 2003 in Hong Kong generated widespread public fear and escalated the efforts of government and medical institutions for infection control. As such, the role of the government and medical institution in preventing public fear and anxiety became prominent. As hypothesized, trust in the institutions is especially relevant to public fear and anxiety during the epidemic crisis. For testing hypotheses involving the role of institutional trust, a telephone survey during the crisis in April 2003 and another telephone survey after the crisis in June 2003 drew data from 9,402 Hong Kong adults for analysis. Results indicate the significant negative effects of trust in the government and trust in the medical institution on anxiety. Supporting the aforementioned hypothesis the effect of institutional trust in the government was significantly more negative on anxiety during the crisis than its aftermath. Hence, the government and medical institution are particularly responsible for anxiety reduction during the crisis. PMID- 19301538 TI - Fatigue among Spanish- and English-speaking Latinos. AB - The present study investigated sociodemographic differences, fatigue severity, and the occurrence of prolonged or chronic fatigue reported by Spanish-speaking and English-speaking Latinos. The sample included 2,102 English-speaking Latinos and 1,348 Spanish-speaking Latinos interviewed as part of an epidemiological study of persons with chronic fatigue syndrome in the Chicago area. Results indicated that English-speaking Latinos scored higher on measure of fatigue than Spanish-speaking Latinos. Further, language status continued to be a predictor of fatigue level even when controlling for other sociodemographic differences found between the groups. Findings suggest that language spoken in Latino populations is important in predicting fatigue, and point to the potential importance of cultural factors such as acculturation or acculturative stresses. PMID- 19301539 TI - Hispanics' locus of control, acculturation, and wellness attitudes. AB - There is reason to believe that various cultural attitudes and beliefs influence certain health behaviors, and additional research should identify the causes of such behaviors. This study explored the relationships among cultural identity, acculturation, locus of control, and health beliefs using a sample of 110 Hispanic individuals taking college classes in the southern or southwestern United States. Path analysis indicated that an external locus of control was positively related to health barrier perceptions and that acculturation was negatively related to health barrier perceptions. The findings suggest that Hispanics' perceived control over health outcomes and positive health beliefs could be enhanced with culturally perceptive counseling. PMID- 19301540 TI - Editorial: the implications of social work practice cost outcomes for American health policy. PMID- 19301541 TI - Understanding cost-effectiveness research applied to social work. AB - This article provides an overview of how cost-effectiveness research (that is more often done in public health settings) can be applied to social work activities. The article describes the degree to which these techniques should influence decision making, the roots of the science for this type of analysis, the meanings of terms that are used, some basic methods, and the next steps for applying this type of work to social work activities. PMID- 19301542 TI - Articulating the evidence base for effective social work practices: building a database to support a geriatric social work policy agenda. AB - In an era that demands accountability and cost-effectiveness, health care and social service professionals are increasingly expected to demonstrate the effectiveness of their interventions and treatments. Social workers, specifically those serving the elderly, have been markedly slow in embracing the need to measure outcomes and produce evidence of their practices, which stymies efforts at advocacy and policy development for the profession. Using a modified systematic review method, the New York Academy of Medicine's (NYAM's) Center for Aging Policy created an evidence database comprising peer-reviewed journal articles on social work interventions and outcomes research on cost-effectiveness and other measures of effectiveness. The evidence database, though focused on care of the elderly, draws on research studies involving age groups across the lifespan. A key product of this initiative is a series of white papers on cost effective interventions in areas such as care management. A work group of social work professionals and a team of NYAM staff with diverse expertise reviewed articles based on their content, key topics, and outcomes. This article describes the steps taken, from the conceptualization of the review process to the development and implementation of the Web-based interface for user access to the evidence database. Though originally conceived as a tool for supporting a policy agenda for the Center for Aging Policy, the evidence database is also a useful reference tool for social workers, policy makers, and others interested in quality of care for elderly. PMID- 19301543 TI - Calculating child welfare separation, replacement, and training costs. AB - Across the country, incoming child welfare caseworkers have an average tenure of 2 years, and states are being evaluated on the quality of caseworker training programs and their ability to provide child welfare services that meet federally mandated outcomes. Policy makers do not know the cost of workforce turnover and replacement or the cost of training child welfare workers. The objectives of the study were to (1) obtain the separation, replacement, and training (SRT) costs for four counties in a major Northeastern state; (2) compare the SRT costs for the three different child welfare pre-service training models; (3) incorporate hidden intercounty transfer and tuition reimbursement cost; and (4) extrapolate the four-county average SRT costs to statewide average SRT costs. Results revealed that one pre-service training model was more than the state tuition for a 4-year bachelor's degree. Further, additional costs were incurred if new caseworkers transferred to another county and/or used Title IV-B tuition reimbursement funds in the first few years of employment. PMID- 19301544 TI - The cost and benefits of cash and services to kinship caregivers. AB - The federal government provides payments to foster parents and licensed relatives who care for abused and neglected children through Title IV-E funds. However, unlicensed relatives must seek assistance through another federal program, Temporary Assistance for Needy Families, which provides lesser amounts of assistance than provided to foster parents and licensed relatives. To redress differences, many states provide supplemental assistance to relative caregivers. This paper profiles one Northeastern state's approach to caregiver rate disparities. The profiled Northeastern state contracted with 17 nonprofit agencies to provide 19 social services. The amount paid for services increased the total public assistance to relative caregivers but also caused licensed relative caregivers to receive more than all other types of caregivers receiving public assistance, thus continuing not eliminating caregiver rate disparities. In addition, services increased during the contract period but decreased again after the contract ended. Further, contract amounts were reduced each year of a 3-year contract. Policy implications for child safety, well-being, and placement permanence are discussed. PMID- 19301545 TI - Cost-utility analysis of high- vs. low-intensity home- and community-based service interventions. AB - Home- and community-based services (HCBS) have been advocated as a mechanism to delay institutionalization and reduce health care costs for the growing senior population. Studies of costs to date have found little evidence of cost savings from HCBS. However, HCBS can be thought to have two main benefits: delaying institutionalization and improving quality of life. Since cost and quality of life can be considered simultaneously in a cost-effectiveness analysis, an exploratory study was conducted to examine the relative cost-effectiveness of a high-dosage (i.e., high-intensity) HCBS intervention (i.e., 1915c Medicaid waiver) compared to a lower-dosage HCBS intervention (i.e., in-home aide service) using quality-adjusted life years as the measure of effectiveness. Findings indicated that high-dosage HCBS is not a cost-effective alternative. The low dosage alternative allows for greater equity through provision of service to a larger pool of individuals in need. PMID- 19301546 TI - Perspectives on the roles and value of social work practice in neighborhood health centers and implications for the reimbursement of services. AB - In recent years, increasing attention has been paid to the delivery of primary care services to underserved populations. This paper presents a subset of data from a larger exploratory study that examined how three professional groups (social workers, physicians, and administrators) in neighborhood health centers perceived the type and extent of psychosocial problems in their patient populations and the roles of their staff in addressing these problems. We examined the perceptions of physicians, administrators, and social workers as to who handles various psychosocial problems and whose responsibility it is to perform a range of psychosocial functions, with a focus on the function of social work. Social workers were highly praised and valued by administrators and physicians. While the respondents in neighborhood health centers confirmed the value of social work, public and private funders are reluctant to reimburse for social services beyond limited clinically diagnosed mental health services. Outcomes studies focused on the efficacy and efficiency of social work practice in neighborhood health centers are necessary to begin to overcome this barrier to social work services. PMID- 19301547 TI - Survival and reoperation risk following bicuspid aortic valve-sparing root replacement. AB - BACKGROUND AND AIM OF THE STUDY: The mid-term outcome of aortic valve-sparing root replacement, reimplanting native bicuspid versus tricuspid aortic valves, is unclear. METHODS: The records of 84 consecutive patients (66 men, 18 women; mean age 46.6 years) who had undergone aortic valve-sparing root replacement using the reimplantation technique between January 1997 and July 2006, were reviewed. Among the patients, 26 (31%) had Marfan syndrome and 14 (17%) had a bicuspid aortic valve. The mean follow up period was 4.1 years. RESULTS: There were three early deaths (4%) and five late deaths (6%). Late survival (>30 days) at five and eight years was 95% and 88%, respectively. The presence of significant coronary artery disease (p = 0.001), a lower preoperative ejection fraction (p = 0.03) and older age (p = 0.04) were the only univariate predictors of death. Freedom from moderate-severe or severe (grade 3 or 4) aortic valve regurgitation at one and five years was 91% and 77%, respectively. Ten patients (12%) required aortic valve reoperation at a mean of 3.4 years after surgery (3/14 bicuspid, 7/70 tricuspid); however, only one reoperation was required among patients undergoing the initial surgery after the year 2000. Freedom from aortic valve reoperation at one and five years was 95% and 83%, respectively. The predischarge degree of aortic valve regurgitation was the sole univariate factor associated with aortic valve reoperation (p = 0.008). Bicuspid valve morphology was not a predictor of either recurrent aortic valve regurgitation, nor of aortic valve reoperation. CONCLUSION: Bicuspid aortic valves may be safely spared by reimplantation during replacement of the aortic root, with similar mid-term durability as for tricuspid aortic valves. PMID- 19301548 TI - Disproportionate shortening of diastolic time and cardiac performance in asymptomatic patients with aortic regurgitation. AB - BACKGROUND AND AIM OF THE STUDY: Exercise-induced abnormalities of the systolic diastolic phase proportion and their impact on hemodynamic variables have been investigated only minimally in asymptomatic patients with chronic aortic regurgitation (AR). METHODS: A group of 33 asymptomatic patients with severe AR and preserved left ventricular (LV) systolic function was studied during incremental exercise, using a combined hemodynamic-radionuclide angiographic approach. The phases of the cardiac cycle were derived with high temporal resolution from the LV time-activity curve. The loss of diastolic time per beat (LDT) was quantified using a regression equation obtained from a healthy control group (n=26). Based on the median LDT at peak exercise, patients were allocated to two groups with LDT < or = 12.4 ms (group A) and LDT > 12.4 ms (group B). RESULTS: At peak exercise the relative duration of LV diastole was significantly shorter in AR patients than in controls (31.4 +/- 3.0 versus 33.1 +/- 2.4 s/min; p = 0.02) and a significant LDT (12.1 +/- 19 ms; p = 0.003) was observed. Group A patients had a higher peak cardiac output (9.2 +/- 2.0 versus 7.4 +/- 2 l/min/m2; p < 0.03), a longer exercise duration (18 +/- 5 min versus 13 +/- 6 min; p < 0.02) and a lesser extent of mean pulmonary artery pressure rise (27 +/- 10 versus 34 +/- 12 mmHg; p = 0.03) than group B patients. CONCLUSION: Cardiac exercise performance in asymptomatic patients with AR is influenced not only by the ability of the cardiovascular system to favorably redistribute total stroke volume, but also to handle volume overload without changing the systole and diastole phase proportions. PMID- 19301549 TI - Effects of patient-prosthesis mismatch on postoperative early mortality in isolated aortic stenosis. AB - BACKGROUND AND AIM OF THE STUDY: Small valve size and patient-prosthesis mismatch (PPM) generate high postoperative transvalvular gradients and may decrease both early and long-term survival. The study aim was to evaluate whether mismatch affected early mortality after aortic valve replacement (AVR) for isolated aortic stenosis (AS). METHODS: A total of 701 patients (437 males, 264 females; mean age 53.3 +/- 15.1 years; range: 14-84 years) with pure AS underwent AVR at the authors' institution between 1985 and 2005. The majority of patients (92%) received a mechanical valve. PPM was considered severe if the indexed effective orifice area was < or =0.65 cm2/m2, and moderate if > 0.65 but < or = 0.85 cm2/m2. RESULTS: Moderate-severe PPM was present in 47% of patients, and severe PPM in 13%. The early mortality was 5.4% (n=38). Multivariate analysis revealed age > or = 70 years (p < 0.001), female gender (p = 0.04) and severe PPM (p = 0.003) as independent predictors of early mortality. Moderate mismatch was not a predictor of early mortality on both univariate and multivariate analysis. Left ventricular dysfunction (ejection fraction < or = 40%) was a risk factor for early mortality only in patients with severe PPM. CONCLUSION: Patient-prosthesis mismatch should be prevented in patients undergoing AVR for isolated AS, especially in those with left ventricular dysfunction. PMID- 19301550 TI - Percutaneous mitral valve dilatation: single balloon versus double balloon. A finite element study. AB - BACKGROUND AND AIM OF THE STUDY: Percutaneous mitral valve (MV) dilatation is routinely performed for mitral stenosis using either a single balloon (SB) or double balloon (DB) technique. The study aim was to compare the two techniques using the finite element (FE) method. METHODS: An established FE model of the MV was modified by fusing MV leaflet edges at commissure level to simulate a stenotic valve (orifice area = 180 mm2). FE models of a 30 mm SB (low-pressure, elastomeric balloon) and an 18 mm DB system (high-pressure, non-elastic balloon) were created. RESULTS: Both, SB and DB simulations, resulted in the splitting of commissures and consequent relief of stenosis (final MV areas of 610 mm2 and 560 mm2, respectively). Stresses induced by the two balloon systems varied across the valve. At full inflation, SB showed a higher stress in the central part of the leaflets and at the commissures compared to DB simulation, which demonstrated a more uniform stress distribution. This was due to mismatch of the round shape of the SB within an oval mitral orifice. Due to its high compliance, commissural splitting was not easily accomplished with the SB. Conversely, the DB guaranteed commissural splitting, even when a high force was required to break the commissure welds. CONCLUSION: The FE model demonstrated that MV dilatation can be accomplished by both SB and DB techniques. However, the DB method resulted in a higher probability of splitting the fused commissures, with less potential for damage to the MV leaflets by overstretching, even at higher pressures. PMID- 19301551 TI - Early hazards of mitral ring annuloplasty in patients with moderate to severe ischemic mitral regurgitation undergoing coronary revascularization: the importance of preoperative myocardial viability. AB - BACKGROUND AND AIM OF THE STUDY: The impact of adding mitral ring annuloplasty (MRA) to coronary bypass grafting (CABG) in patients with ischemic mitral regurgitation (iMR) is unclear. The study aim was to compare the 30-day and four year survival of patients with moderate to severe iMR undergoing CABG or CABG+MRA, and to investigate the role of contractile reserve (CR) in the prognostic response to MRA. METHODS: A total of 76 coronary patients (61 men, 15 women; mean age 62 +/- 9 years) with poor left ventricular ejection fraction (LVEF) of 33 +/- 11% and grade > or =2 iMR underwent low-dose dobutamine echocardiography to identify their CR before CABG. The survival of 34 patients undergoing CABG+MRA was compared to that of 42 patients who underwent CABG alone. The groups were further substratified according to their preoperative CR. RESULTS: During follow up, 24 patients died from cardiac causes, and two others required heart transplantation. At one year, the residual iMR and NYHA functional class were lower in patients undergoing MRA than in those that did not. The 30 day and four-year survivals were lower in patients undergoing MRA in the absence of CR than in the other patients (71 +/- 11% versus 95 +/- 3% at 30 days, p = 0.002; 35 +/- 11% versus 69 +/- 6% at four years, p = 0.008). Cox's proportional hazard analysis identified CR (HR = 0.14, 95% CI 0.05-0.38, p < 0.001), MRA (HR = 3.54, 95% CI 1.48-8.50, p = 0.004), additive EuroSCORE (HR = 1.29, 95% CI 1.08 1.55, p = 0.006) and LVEF (HR = 0.93, 95% CI 0.59-0.98, p = 0.001) as independent predictors of long-term outcome in this population. CONCLUSION: In patients with moderate to severe iMR, survival after CABG is mainly influenced by the presence of CR. By contrast, adding MRA to CABG does not affect long-term survival, except in patients without CR, in whom it increases early mortality. PMID- 19301552 TI - Autocrine serotonin and transforming growth factor beta 1 signaling mediates spontaneous myxomatous mitral valve disease. AB - BACKGROUND AND AIM OF THE STUDY: Although serotonin and serotoninergic drugs are known to cause myxomatous-like valvulopathy, the role of serotonin in spontaneous myxomatous valve disease (MVD) remains unclear. Tryptophan hydroxylase 1 (TPH1) is the limiting enzyme for peripheral serotonin synthesis, and its expression in myxomatous valves could implicate an autocrine serotonin signaling mechanism. Studies in cultured cells demonstrate a close coupling between serotonin and transforming growth factor beta1 (TGFbeta1) signaling. The study aim was to investigate serotonin and TGFbeta1 signaling in spontaneous MVD. METHODS: In canine normal and myxomatous mitral valves, target signaling proteins including TPH1, serotonin 2B receptor (5HT(2B)R), serotonin transmembrane transporter (SERT), total and phosphorylated extracellular signaling-regulated kinase (ERK) 1/2, latent TGFbeta1 and TGFbeta1 receptors I and II, were studied using immunohistochemistry and immunoblot analysis. In human myxomatous valves, TPH1 was determined using immunofluorescence and immunoblot analysis. RESULTS: In canine mitral valves, both 5HT(2B)R and TPH1 were increased in myxomatous valves, whereas SERT, a key protein in serotonin metabolism, was decreased in myxomatous valves. Phosphorylated, but not total, ERK 1/2 was increased in myxomatous valves, consistent with an enhanced active serotonin signaling. The expression of TGFbeta1 receptors I and II, and of latent TGFbeta1, was increased in myxomatous valves. Human myxomatous mitral valves expressed TPH1. CONCLUSION: The expression of TPH1 by canine and human myxomatous valves demonstrates a capacity for local serotonin production. Key signaling protein expression patterns support active serotonin and TGFbeta1 signaling in canine myxomatous valves. These findings implicate an autocrine serotonin and TGFbeta1 mechanism in the pathogenesis of spontaneous MVD. PMID- 19301553 TI - Calcification of left-sided valvular structures: evidence of a pro-inflammatory milieu. AB - BACKGROUND AND AIM OF THE STUDY: Calcification of the cardiac valves occurs more frequently on the left side, most commonly involving the aortic and mitral valves. The differences in hemodynamics and blood pressure between the right and left sides have been considered as possible causes. However, the reason for this preferential distribution remains unknown. It is hypothesized that a stronger pro inflammatory milieu exists in the left side of the heart, and might be responsible for valve calcification occurring most frequently on that side. METHODS: An extensive search of the medical literature using PubMed and Google Internet search engines and online databases, as well as textbooks and journals, was conducted. Search areas included the pathophysiology of normal and dystrophic calcification, and the incidence and distribution of native valve involvement in different diseases. RESULTS: The majority of the disease processes affecting cardiac valves have an increased predilection for the left-sided valves. This was also found to be true in systemic diseases with cardiac involvement. Diseases where the right-sided valves are subjected to higher pressure and comparably forceful hemodynamics were found not to demonstrate a similar increase in the incidence of valve calcification. Pulmonary autografts or homografts in the aortic position appear not to demonstrate the same rate of calcification as their aortic counterparts, despite being subjected to the same hemodynamics. CONCLUSION: The reason for valve calcification occurring more frequently on the left side remains unknown. All available evidence does not support the proposal that a higher pressure and more forceful hemodynamics are causes for this observation. Rather, the data indicate that there is a pronounced pro inflammatory milieu in the tissues of the left-sided cardiac valves, that may be responsible for such increased valve calcification. Further investigations are required in order to assess, quantitatively, the differences in inflammatory response between right- and left-sided cardiac valves. PMID- 19301554 TI - Natural history of aortic valve disease following intervention for rheumatic mitral valve disease. AB - BACKGROUND AND AIM OF THE STUDY: A significant proportion of patients who require interventions for rheumatic mitral valve (MV) disease have coexisting aortic valve (AV) disease. To date, little is known of the natural history of AV disease in these patients. METHODS: The details of a cohort of 200 patients (146 females, 54 males; mean age at MV intervention 30.3 +/- 9.9 years) with rheumatic heart disease were retrospectively reviewed. The patients had undergone an index MV intervention (either closed or balloon mitral valvotomy) or MV replacement between 1994 and 1996, and received long-term regular follow up examinations. The clinical and echocardiographic data at entry and at follow up were noted. Patients were allocated to two groups, based on whether the AV disease was absent (group I, n=98) or present (group II, n=102) at baseline. The AV disease was categorized as thickening only (group IIA), isolated aortic regurgitation (AR) (group IIB), or combined aortic stenosis (AS) and AR (group IIC). No patient had isolated AS at baseline. RESULTS: The mean follow up period was 9.3 +/- 1.07 years; during which 10 patients in group I developed new AV disease, which included AV thickening only (n=2), trivial-mild AR (n=7) and mild AS with trivial AR (n=1). Of 16 patients in group IIA, 11 developed isolated AR, and one patient progressed to have mild AS and AR. Among 69 patients in group IIB, 22 (31.9%) developed AS, and all had either mild (n=8) or moderate (n=14) AR with mild AS. Group IIC included 17 patients with mild combined AV disease at baseline, except for moderate AS and moderate AR in one patient each. Among 16 patients with mild AS in group IIC, six progressed to moderate AS and two to severe AS. AR became moderate in 10 patients and severe in one patient. The two patients who progressed to severe AS requiring AV replacement had mild AS at baseline. No patient who developed new combined AV disease had lesions with severity more than mild AS or moderate AR. On logistic regression analysis of the variables predisposing to progression of AV disease, such as age, gender, history of rheumatic fever (RF) and recurrence, and interval from RF episode to symptom onset, only the initial AV gradient was identified as being statistically significant (beta coefficient 0.528, SE = 0.17, p < 0.0001). CONCLUSION: Patients with no or mild AV disease at the time of MV intervention rarely develop severe AV disease, and seldom require AV surgery over the long-term follow up. The presence of mild AS at baseline is predictive in the minority of cases where AV disease will progress relatively more rapidly. PMID- 19301555 TI - The spectrum of rheumatic heart disease in the southeastern Anatolia endemic region: results from 1900 patients. AB - BACKGROUND AND AIM OF THE STUDY: Rheumatic heart disease (RHD) continues to affect mainly young adults, and accounts for a major proportion of all cardiovascular diseases in the Southeastern Anatolia region, where the disease is still endemic. The study aim was to determine the echocardiographic profile of RHD in this region, from which no recent data are available. METHODS: In this retrospective study, transthoracic echocardiographic data acquired between June 2003 and January 2008 were reviewed. Information was gathered from the database of the authors' echocardiography laboratory, which included age, gender, clinical diagnosis and echocardiographic findings. In patients with more than one echocardiographic record, only the first echocardiographic data were included in the study. RESULTS: A first admission with full echocardiographic data of RHD was found in 1,900 cases among 43,900 subjects screened (4.3%). Of the 1,900 subjects, 537 (28%) were male (mean age: 39.7 +/- 15.7 years) and 1,363 (72%) were female (mean age: 41.3 +/- 13.7 years). There was no gender difference between mixed valvular lesions. Severe mitral stenosis (MS) was identified in 144 subjects. Males were affected more severely than females, provided that adjustments were made according to valve area < or =1 cm2 or >1 cm2. A total of 594 subjects had MS with a valve area < or =1.5 cm2. Severe aortic regurgitation (AR) (grade > or =3) was more common in males than in females. The echocardiographic scores showed 525 subjects (28%) to have mild, 1083 (57%) to have moderate, and 292 (15%) to have severe AR. Among a total of 1163 patients with MS, 385 (33%) had > or =0.5 cm nodular calcification on their valves. CONCLUSION: Currently, RHD remains an alarming and unresolved health problem in the Southeastern Anatolia region. While almost 75% of affected subjects were female, males were more severely affected. In addition, subjects were relatively old, and most were affected by mixed valvular disease of an advanced stage. PMID- 19301556 TI - Taking patient mortalities into account when assessing change in NYHA classification from baseline. AB - Patient response to treatment is often monitored by comparing metrics of performance immediately before treatment and at follow up. One such metric in patients with heart disease is the New York Heart Association (NYHA) classification, which is graded on a I to IV scale. In order for such an analysis to be unbiased, any missing NYHA classifications at follow up must be 'missing completely at random'. The study aim was to determine how the exclusion of expired patient data may bias the analysis of changes in NYHA class, with attention focused on the effects of ignoring patient mortalities, not only for the sake of simplicity but also that this might inflict the largest degree of bias in favor of the treatment. PMID- 19301557 TI - Ross and Ross-Konno procedures in infants, children and adolescents: a 13-year experience. AB - BACKGROUND AND AIM OF THE STUDY: Due to hemodynamic performance and potential for growth of the pulmonary autograft, the Ross operation is considered to be the surgery of choice for irreparable aortic valve disease in pediatric patients. The study aim was to analyze the long-term clinical and echocardiographic results of the Ross operation. METHODS: Between February 1993 and July 2006, 55 consecutive patients (mean age 10.0 +/- 6.2 years; range: 3 months to 18 years) underwent eithera Ross operation (n=46) or a Ross-Konno procedure (n=9). The underlying left ventricular outflow tract pathology was mainly congenital (n=47). Among patients, 23 (42%) had undergone a previous aortic valve procedure. Concomitant procedures were performed in 16 patients (29%). The Ross operation was performed as a root replacement in all cases; the mean cross-clamp time was 132 min (range: 100-188 min). The autograft diameter was indexed to the body surface area and compared to normal values. The mean follow up was 5.5 +/- 3.8 years, and was 100% complete. RESULTS: There was one early death (2%) and two late deaths (4%). The actuarial patient survival was 93% at 10 years. None of the patients developed moderate or severe autograft regurgitation. All measured maximal root diameters were above the 90th percentile of normal aortic diameter, without correlation to autograft regurgitation. Five patients (9%) had a mean homograft gradient > or = 40 mmHg, and two (4%) were reoperated on. The freedom from reoperation for homograft degeneration was 91% at 10 years. CONCLUSION: Autograft regurgitation after the Ross and Ross-Konno procedures is uncommon, and the risk of homograft degeneration appears low. Autograft dilatation is common but does not correlate with autograft regurgitation. When considering long-term freedom from autograft and homograft degeneration, the results of the present study confirm the Ross operation as the surgery of choice for irreparable aortic valve disease in infants, children and young adults. PMID- 19301559 TI - Hemodynamic characterization of the Sorin Mitroflow pericardial bioprosthesis at rest and exercise. AB - BACKGROUND AND AIM OF THE STUDY: Although the long-term clinical outcome after aortic valve replacement (AVR) with the Sorin Mitroflow pericardial bioprosthesis has been well described, few data are available with regards to hemodynamic performance. On the basis of its specialized design, with the pericardium mounted on the outside of the stent, the Mitroflow valve is considered to provide optimal opening and orifice areas. METHODS: Between February 2006 and April 2007, a total of 127 patients (66 females, 61 males; mean age 77.6 +/- 5.2 years) underwent AVR with the Mitroflow valve at the authors' institution. Echocardiography was performed at discharge in 91 patients, while a six-month follow up examination was conducted in 78 (including ergometer hemodynamics in 25 cases). RESULTS: The mean systolic pressure gradient (MPG) ranged from 19.7 +/- 3.3 mmHg (size 19 valve) to 12.4 +/- 2.4 mmHg (size 27 valve), and the mean effective orifice area (EOA) from 0.96 +/- 0.0 cm2 (size 19 valve) to 2.36 +/- 0.4 cm2 (size 27 valve). The incidence of mild-to-moderate and severe prosthesis-patient mismatch (PPM) was 33% and 10% at the six-month follow up. During exercise (25 W-100 W), the MPG increased from 17.1 +/- 3.6 mmHg to 23.6 mmHg in valve sizes 19 to 21, and from 12.2 +/- 3.6 mmHg to 15.9 +/- 2.5 mmHg in valve sizes 23 to 27. The effective orifice fraction (EOF = EOA/annulus area) was 38 +/- 7%. CONCLUSION: The Mitroflow valve exhibits an adequate MPG and EOA, as might be expected for a pericardial bioprosthesis. Of note, the gradient increase during exercise was low, especially for the larger valve sizes. Due to its special design, the Mitroflow prosthesis shows a large opening, as demonstrated by the high EOF. These data relating to the EOA of all prosthesis sizes may help surgeons to select the minimum prosthesis size in order to prevent PPM. PMID- 19301560 TI - Does sodium dodecyl sulfate wash out of detergent-treated bovine pericardium at cytotoxic concentrations? AB - BACKGROUND AND AIM OF THE STUDY: The ionic detergent sodium dodecyl sulfate (SDS) is a proposed treatment for the removal of antigenic proteins from unfixed biological scaffolds used in tissue engineering. However, questions remain about possible cytotoxic effects of SDS-treated tissues. The study aims were to: (i) develop a sensitive SDS assay for physiological solutions; (ii) measure SDS concentrations in the washing media of SDS-treated tissue; and (iii) determine cytotoxic SDS concentrations in cultured ovine vascular cells. METHODS: An assay was developed to monitor SDS concentrations at microM levels, based on attenuated total reflectance infrared spectroscopy. Bovine pericardium was treated with SDS (1.0 to 0.01%) and washed for 96 h. The SDS concentration in the washing media was measured at 24-h intervals; data were expressed as microM/g tissue. Ovine vascular cells were cultured in DME media at 37 degrees C for 48 h in various SDS concentrations (10 to 1000 microM). The cells were then counted, and the percentage live cells expressed, based on trypan blue exclusion (n=5). RESULTS: SDS concentrations > or =10 microM significantly reduced (p < 0.05) the total cell number, while concentrations > or =100 microM reduced (p < 0.05) the percentage live cells of ovine vascular cell cultures. SDS was present in the washing media of SDS-treated bovine pericardium. SDS leaching from bovine pericardium was found to depend on the SDS concentration used for the treatment, and diminished with time. CONCLUSION: SDS leaches from SDS-treated bovine pericardium at concentrations that are potentially cytotoxic. An understanding of the dynamics of SDS washout, based on a sensitive SDS assay, may lead to the creation of protocols for the preparation of biological scaffolds that are free from cytotoxic leaching. PMID- 19301558 TI - Thirteen years' experience with the Ross Operation. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate 13 years' results of the Ross operation, with special attention paid to the late function of the pulmonary autograft (PA) and the right ventricular outflow tract (RVOT) allograft, in an attempt to identify risk factors that might significantly influence their late performance. METHODS: Between May 1995 and March 2006, a total of 272 patients (mean age 30 +/- 11 years) was submitted to a Ross operation at the authors' institution. The most prevalent etiology was rheumatic disease (47%). The PA was implanted as a root replacement in 236 cases, and as an intraluminal cylinder in 36. The RVOT was reconstructed with a cryopreserved allograft in conventional fashion (n=142), with a proximal extension of the allograft with pericardium (n=46), with an allograft decellularized with deoxycholic acid (n=39), or an allograft decellularized with sodium dodecylsulfate (SDS) (n=44). The mean follow up was 67 months (range: 1-153 months: the total follow up was 1,525 patient-years. RESULTS: Hospital mortality was 2.9% and late survival 93% at 12 years. There were two episodes of cerebral thromboembolism, and six patients developed bacterial endocarditis. Sixteen patients required reoperation for problems involving the PA and/or the RVOT allograft, progression of rheumatic mitral valve disease and iatrogenic coronary insufficiency. After 12 years, 97% and 95% of patients were free from reoperation with the PA and allograft, respectively. Over time, the PA showed increasing dimensions, and 10 patients had a diameter >45 mm. Univariate analysis revealed degenerative disease and aortic annulus >25 mm as significant risk factors for late PA dilatation. Female gender and bicuspid aortic valve were borderline factors for this type of complication. Another 14 patients had late moderate PA insufficiency, and this was correlated with a preoperative diagnosis of aortic insufficiency. Reconstruction of the RVOT with allografts decellularized with SDS were associated with lower gradients at late follow up. CONCLUSION: Late results with the Ross operation were associated with excellent long-term survival and a low incidence of reoperations up to 13 years. Due to late PA dilatation and/or progressive valvar insufficiency, some reoperations may be expected with a longer follow up. For the RVOT reconstruction, the use of decellularized allografts with SDS may prove to be a good alternative to RVOT reconstruction. PMID- 19301561 TI - Echocardiographic measurement of mitral intertrigonal distance is an adjunct to annuloplasty ring sizing. AB - BACKGROUND AND AIM OF THE STUDY: Annuloplasty sizing with standard valve sizers may be imprecise and difficult in minimally invasive procedures. It is hypothesized that a constant clinical conversion factor relates the echocardiographic aortic annulus diameter (AAD) and the intertrigonal distance (ITD) in patients with degenerative mitral regurgitation (MR). This may provide another method to size the annuloplasty ring required for mitral valve repair. METHODS: An observational study of 50 patients with degenerative MR undergoing robotic-assisted surgery was conducted. All patients underwent surgery between September 2005 and November 2007. The AAD at the base of the aortic leaflets was measured using intraoperative two-dimensional transesophageal echocardiography. The ITD was measured independently under direct vision during surgery. The echocardiographic ITD was then determined by dividing the AAD by 0.8, and the value for each patient compared to the corresponding surgical measurement. Agreement was assessed statistically using the Bland-Altman method. RESULTS: The limits of agreement were -3 mm (t = 2.010; 49 df; p = 0.05; 95% CI: -4 to -2 mm) to 3 mm (t = 2.010; 49 df; p = 0.05; 95% CI: 2 to 4 mm). In 86% of cases (43/50), the differences between the two methods was < or = 2 mm. CONCLUSION: In most cases of degenerative mitral valve disease the echocardiographic ITD measurement is clinically acceptable, and may serve as an adjunct to existing methods when sizing the annuloplasty ring required for repair. PMID- 19301562 TI - Clinical outcome of a simplified technique for aortic valve replacement with stentless bioprostheses. AB - BACKGROUND AND AIM OF THE STUDY: Aortic stentless bioprostheses provide good clinical and hemodynamic results, but may be difficult to implant. Their use is also contraindicated in the presence of a severely calcified aortic root. The study aim was to assess the mid-term results of a simplified implant technique of the Sorin Pericarbon Freedom stentless bioprosthesis (SPF), that allows its use in the presence of severe aortic root calcification. METHODS: Between 2001 and 2007, a total of 51 patients underwent aortic valve replacement (AVR) with the SPF, using a new technique which consisted of standard annular fixation and the fixation of each strut with a single 'U' stitch. The perioperative characteristics, early and late mortality and occurrence of valve-related events were evaluated. The overall mean cross-clamp and cardiopulmonary bypass times were 91.5 +/- 30.2 and 125.3 +/- 44.9 min, respectively, and 77.8 +/- 17.7 and 105.6 +/- 22.8 min, respectively, for AVR (these times were comparable to those required in patients receiving stented valve bioprostheses). The mean follow up period was 40.5 months (range: 3-75 months), and was 100% complete. RESULTS: Hospital mortality was 4% and survival 76.5 +/- 14.5% at six years. Late deaths occurred in eight patients; in four cases this was valve-related (1.9%/patient year). Freedom from valve-related death and reoperation was 91 +/- 9% and 98 +/- 2% respectively, at six years. The mean transprosthetic gradients were 12.0 +/- 3.4, 9.9 +/- 2.6, 8.7 +/- 4.2, 5.2 +/- 3.9, 4.5 +/- 3.0 and 3.2 +/- 2.7 mmHg for the 19, 21, 23, 25, 27 and 29 mm valve sizes, respectively. No valvular or paravalvular regurgitation was recorded. CONCLUSION: This new implantation technique for the aortic SPF stentless bioprosthesis is simple, effective and versatile, and showed good early results. It may be applicable to other stentless bioprostheses, and also be particularly useful in patients with small aortic annulus and in those with a calcified aortic root. PMID- 19301563 TI - Triangular resection for posterior mitral prolapse: rationale for a simpler repair. AB - Patients with mitral regurgitation caused by posterior leaflet prolapse are ideal candidates for valve repair, yet all too often these patients receive prosthetic replacement. Surgeon discomfort with available repair techniques is often cited as the reason for this suboptimal result. The dissemination of simpler repair techniques might increase the proportion of patients receiving mitral valve repair rather than replacement. Triangular posterior leaflet resection is an easy, effective and durable method for correcting posterior leaflet prolapse. The combination of triangular resection and annuloplasty is an excellent option for mitral valve repair in most patients with posterior leaflet prolapse. PMID- 19301564 TI - Aortic valve regurgitation in a patient affected by KBG syndrome. AB - The KBG syndrome is a very rare condition characterized by developmental delay, short stature, distinct facial dysmorphism, macrodontia of the upper central incisors and skeletal abnormalities. Associated congenital heart defects have been described in 9% of patients. Herein is described a case of aortic root dilatation with significant regurgitation in a young patient affected by KBG syndrome. Surgical inspection showed a dilated aortic annulus, slightly dilated aortic sinuses, a tricuspid valvb with slightly thickened cuspal margins and central regurgitation. Histological examination showed a fibrous hyaline involution of the valvular leaflets. To the authors' knowledge, this is the first reported case of KBG syndrome affected by aortic root dilatation with severe regurgitation. Morphology of the aortic valve leaflets was relatively normal, but the annulus was dilated in the absence of any history of rheumatic fever, hypertension, connective tissue or rheumatic systemic diseases. The unusual findings in this young patient raised questions regarding the as-yet unexplained etiopathogenesis of the KBG syndrome. PMID- 19301565 TI - Aortic stenosis five years after coronary artery bypass grafting in a young patient with homozygous familial hypercholesterolemia. AB - Homozygous familial hypercholesterolemia (FH) is an autosomal dominant disease caused by a mutation in the low-density lipoprotein (LDL) receptor. This mutation can lead to increased serum LDL, and subsequently to premature coronary artery disease. It may also lead to valvular and supravalvular aortic stenosis, these complications being cardinal in the natural course of the disease. The surgical treatment of aortic stenosis in patients with homozygous FH is accompanied by high risk, even in skillfu1 hands. Herein is presented the long-term follow up of a young patient with homozygous FH who underwent coronary artery bypass graft surgery at the age 14 years. Although the patient developed aortic stenosis five years later, neither the native coronary vessels nor grafted vessels underwent any atherosclerotic changes during this period. PMID- 19301566 TI - Preterm birth prevention by 17 alpha-hydroxyprogesterone caproate vs. daily nursing surveillance. AB - OBJECTIVE: To compare the incidence of spontaneous recurrent preterm delivery (SPTD) between women receiving 17 alpha-hydroxyprogesterone caproate (17P) and women receiving daily perinatal nursing surveillance (dPNS) with home uterine activity monitoring. STUDY DESIGN: Women enrolled for dPNS or weekly nursing visits with 17P injection were eligible. Included were singletons with previous SPTD, without preterm labor (PTL), cerclage or vaginal bleeding and < 27 weeks at enrollment. 17P and dPNS patients were matched 1:1 by race, marital status, tobacco use and number of SPTDs. Primary study outcome was incidence of spontaneous PTD. RESULTS: Data from 342 matched pairs were compared. Diagnosis of PTL (39.2% vs. 60.8%) and tocolytic use (12.9% vs. 49.7%) was decreased with 17P vs. dPNS (p < 0.001). The incidences of spontaneous PTD at < 32, 35 and 37 weeks were similar between the groups. CONCLUSION: There was no difference in recurrent SPTD between women treated with 17P and those receiving dPNS. PMID- 19301567 TI - Postcesarean pelvic floor dysfunction contributes to undisclosed psychosocial morbidity. AB - OBJECTIVE: To estimate the prevalence and severity of postcesarean pelvic dysfunction. STUDY DESIGN: Using biopsychosocial interviewing at home, 184 postcesarean primiparas were compared to 100 vaginally delivered women regarding symptoms of stress incontinence, anal incontinence and dyspareunia. Delivery details were confirmed from medical records. RESULTS: Comparison of postcesarean vs. vaginally delivered women revealed stress incontinence in 33% vs. 54% and dyspareunia in 27% vs. 46%, both differences reaching statistical significance, unlike anal incontinence, which was manifest in 51% vs. 44%. When compared to emergency cesarean the relative risk of stress incontinence following an elective cesarean was 0.99 (0.71, 1.39), of dyspareunia 1.02 and of anal incontinence 1.05, indicating no statistically significant difference. Thirty (22%) stress incontinent and 4 (3%) fecally incontinent mothers used pads continuously, suggesting severe physical morbidity. Severe dysphoria (depression) was expressed by 41 (35%) stress incontinent mothers, 38 (30%) with dyspareunia and 34 (26%) with anal incontinence; the association of severe dysphoria with dyspareunia was statistically significant (OR = 2.504 [1.362, 4.602]). Few women came forward to seek help. CONCLUSION: Pelvic dysfunction was similar after elective or emergency cesarean. Compared to vaginal delivery, postcesarean stress incontinence and dyspareunia were less frequent but biopsychosocial morbidity could be severe. PMID- 19301568 TI - Vulvar diseases of the tropics: a review. AB - With the increasing immigrant population in the United States, it is imperative for clinicians in the United States to become versed in tropical vulvar conditions uncommon to indigenous members of developed nations that may present to their practice. Travelers to tropical areas may also contract diseases that occur in these areas, which may manifest as a vulvar condition and mimic a sexually transmitted disease. A brief review is presented. PMID- 19301569 TI - Lower chance of pregnancy with repeated cycles with in vitro fertilization. AB - OBJECTIVE: To evaluate pregnancy outcomes in initial and replicate IVF cycles. STUDY DESIGN: A retrospective analysis of 2,167 fresh, nondonor IVF cycles performed in a large private practice from January 1, 2005, to March 1, 2006. Standard controlled ovarian hyperstimulation and laboratory protocols were followed. RESULTS: Patients undergoing multiple treatment cycles were significantly older. There was no difference in body mass index or percentage of cancelled cycles with increasing number of IVF attempts. The number of retrieved, mature and fertilized oocytes progressively declined as the number of treatment cycles increased. The number of embryos transferred increased with increasing number of treatment cycles. Implantation, pregnancy and clinical pregnancy rates decreased significantly with the second treatment cycle and more markedly with 3 5 treatment cycles. CONCLUSION: The likelihood of a successful outcome declined with each additional treatment cycle. The most notable decrease in clinical pregnancy rates occurred after the third cycle. Patients who fail to conceive after 3 cycles of IVF should be counseled to begin considering other options. PMID- 19301570 TI - Progestins for the prevention of spontaneous preterm birth: review and implications of recent studies. AB - Progesterone plays a central role in the mechanisms of parturition in many species. Despite remarkable advances in our understanding of this hormone's mechanism of action, its roles in human pregnancy maintenance and parturition are not fully appreciated. Proper scientific hypothesis testing of progestins to prevent preterm birth has been limited because of the issues that can plague interventional trials in obstetrics, including patient selection, choice of outcome and power. The largest studies enrolling patients with a history of prior preterm birth alone to prevent recurrence appear contradictory. In contrast, consistent evidence from one multinational trial and a secondary analysis of another suggests cervical length may serve to identify potential responders to this therapy. Finally, the safety of progestin administration is a legitimate concern and a meta-analysis justifies the need for further investigation of safety issues. This review presents recent findings regarding progestin therapy from both clinical and laboratory data and considers unresolved issues for use of these agents. PMID- 19301571 TI - Management of gestational trophoblastic neoplasia with 5-fluorouracil and actinomycin D in northern China. AB - OBJECTIVE: To study risk factors associated with the number of chemotherapy courses required to obtain normal beta-human chorionic gonadotropin (beta-hCG) levels and evaluate the effectiveness of combination chemotherapy using 5 fluorouracil (5-FU) and actinomycin D (Act D) (FA) for gestational trophoblastic neoplasia (GTN). STUDY DESIGN: A retrospective study of GTN cases from January 1995 to July 2007 at the Beijing Obstetrics and Gynecology Hospital was performed. RESULTS: During the study period, 240 cases of GTN were diagnosed and managed at our institution. A stepwise regression analysis revealed that the interval between evacuation and chemotherapy, parity, age and beta-hCG level were associated with number of chemotherapy courses required to obtain normal beta-hCG levels. In addition to 4 patients with placental site trophoblastic tumor, FA was used in 218 patients (136 low risk and 82 high risk). CONCLUSION: Early detection of GTN and timely administration of chemotherapy decreases the number of chemotherapy courses required. The combination chemotherapy of 5-FU and Act D is effective in low- and high-risk patients with GTN, but not in patients with placental site trophoblastic tumor; it can be used as primary chemotherapy regimen in high-risk patients with GTN, except those with extensive metastases. PMID- 19301572 TI - Abortion and the risk of subsequent preterm birth: a systematic review with meta analyses. AB - OBJECTIVE: To conduct a systematic review and meta-analyses of studies that test the association between induced or spontaneous abortion and subsequent preterm birth. STUDY DESIGN: International databases were reviewed (1995-2007) using the terms preterm, premature, birth, labor, delivery, abortion, induced abortion, miscarriage and spontaneous abortion. Only studies that met prespecified objective criteria for methodologic design and reporting were included in the meta-analyses. RESULTS: Twelve induced and 9 spontaneous abortion studies met inclusion criteria. Common adjusted odds ratios (ORs) for preterm birth following 1 and > or = 2 induced abortions were 1.25 (95% confidence interval [95% CI] 1.03 1.48) and 1.51 (95% CI 1.21-1.75), respectively. Four studies provided a common adjusted OR for < or = 32 weeks' births of 1.64 (95% CI 1.38-1.91). Meta regression analysis revealed a previously unrecognized inverse relationship between the In OR and the control population preterm birth rate, explaining in part the observed heterogeneity among studies. Analysis of spontaneous abortion and subsequent preterm birth revealed a similar common adjusted OR and inverse meta-regression on the control preterm birth rates. CONCLUSION: Induced and spontaneous abortion are associated with similarly increased ORs for preterm birth in subsequent pregnancies, and they vary inversely with the baseline preterm birth rate, explaining some of the variability among studies. PMID- 19301573 TI - Predictors of psychological well-being in a sample of women with vulval pain. AB - OBJECTIVE: To investigate factors that predicted mental health-related quality of life in a sample of Australian women with vulval pain. STUDY DESIGN: A convenience sample of Australian women with and without vulval pain completed a questionnaire containing a range of well-validated self-report measures. Questionnaires were returned by mail by 51 women with vulval pain aged between 19 and 68 years and 46 women without vulval pain aged between 21 and 65 years. RESULTS: Women with vulval pain reported significantly worse levels of mental health-related quality of life than women without vulval pain. Thoughts about vulval conditions being serious conditions that had greatly affected the women's lives were more likely to lead to lower quality of life, and believing that vulval pain conditions were likely to have been caused by psychological factors such as stress or worry was also associated with reduced mental health-related quality of life. Adequate levels of social support and helpful emotion-focused coping strategies were positively related to quality of life among women with vulval pain. CONCLUSION: Illness perceptions played an important role in the women's mental-health related quality of life. Appropriate psychosocial interventions targeting particular perceptions may improve quality of life. PMID- 19301574 TI - Pycnogenol: a rapidly expanding role in the management of systemic diseases besides dysmenorrhea. PMID- 19301575 TI - A cranial intercondylar arthroscopic approach to the caudal medial femorotibial joint of the horse. AB - REASON FOR PERFORMING STUDY: Current noninvasive techniques for imaging the soft tissue structures of the stifle have limitations. Arthroscopy is commonly used for the investigation and treatment of stifle pain. Cranial and caudal arthroscopic approaches to the femorotibial joints are used. However, complete examination of the axial aspect of the medial femorotibial joint (MFTJ) is not possible currently. OBJECTIVE: To develop a cranial approach to the caudal pouch of the MFTJ and to assess whether it would allow a more complete examination of the compartment and facilitate the caudomedial approach. METHOD: The regional anatomy was reviewed and the technique developed on cadavers. A series of nonrecovery surgeries were performed to evaluate the procedure, which was then used in 7 clinical cases. Advantages compared to existing techniques and complications encountered were recorded. RESULTS: Successful entry into the caudal pouch of the MFTJ was achieved in 20 of 22 cadaver legs, 8 of 8 joints of nonrecovery surgery horses and 6 of 7 clinical cases operated. The caudal ligament of the medial meniscus could be visualised, along with other axial structures of the caudal joint pouch. The technique was used to facilitate a caudomedial approach and allowed better triangulation within the joint space. Complications were minor and included puncture of the caudal joint capsule and scoring of the axial medial femoral condyle. CONCLUSIONS AND POTENTIAL RELEVANCE: It is possible to access the caudal pouch of the MFTJ arthroscopically using a cranial intercondylar approach. The technique has advantages when compared to existing techniques and is associated with few significant complications. A cranial approach to the caudal pouch of the MFTJ could complement existing techniques and be useful clinically. PMID- 19301576 TI - Prevalence of various radiographic manifestations of osteochondrosis and their correlations between and within joints in Dutch warmblood horses. AB - REASONS FOR PERFORMING STUDY: Osteochondrosis (OC) is the most important orthopaedic developmental disorder in horses and may manifest in several different forms. No detailed study on the prevalence and/or interrelation of these forms is available, even though these data are a prerequisite for conclusive genetic studies. OBJECTIVES: To assess the prevalence of the various manifestations of OC as detected radiographically and to evaluate possible relationships between their occurrence within the same joint and between different joints. METHODS: The FP (femoropatellar), TC (tarsocrural) and MCP/MTP (metacarpophalangeal/metatarsophalangeal) joints of 811 yearlings selected randomly, descending from 32 representative stallions, were radiographed and scored for the presence and grade of osteochondrotic lesions. Results were compared at the sire, animal, joint and predilection site levels. RESULTS: In the FP joint, the percentage of animals showing normal joint contours in all sites was 60.7%. For the TC joint and the combined MCP/MTP joints, these figures were 68.6 and 64.6%, respectively. For all joints combined, the percentage dropped to 30.5%. Sedation improved detection of OC lesions in the FP joint. There was a high correlation between the right and left joints. The correlation between flattened bone contours and fragments was considerably less. CONCLUSIONS: Scoring on a detailed scale is necessary to achieve good insight into the prevalence of OC. Observations on the right and left joints can be combined in further analyses, whereas flattened bone contours and fragments should be evaluated as statistically different disorders. POTENTIAL RELEVANCE: This study provides insight into the prevalences of various manifestations of OC and their relationships, within and between joints. These results form the basis for detailed quantitative and/or molecular genetic studies that should lead to the establishment of breeding indices and/or genetic marker sets for OC. PMID- 19301577 TI - Distal limb cast sores in horses: risk factors and early detection using thermography. AB - REASONS FOR PERFORMING STUDY: There is a lack of evidence-based data on the prevalence, outcome and risk factors of distal limb cast sores, and no objective tool has been described for the early detection of cast sores. OBJECTIVES: To investigate the prevalence, location, outcome and risk factors of cast sores after application of a distal limb cast and to determine whether static thermography of the cast is a valuable tool for the assessment of sores. METHODS: A prospective study was conducted on horses treated with a distal limb cast. At each cast removal, cast sores were graded as superficial sores (SS), deep dermal sores (DS) or full thickness skin ulcerations (FS). In several cases, a thermographic evaluation of the cast was performed immediately prior to removal and differences in temperature (AT) between the coolest point of the cast and 2 cast regions predisposed for sore development (dorsoproximal mc/mtIII and palmar/plantar fetlock) were calculated. RESULTS: Mean +/- s.d. total casting time of 70 horses was 31 +/- 18 days. Overall, 57 legs (81%) developed at least SS. Twenty-four legs (34%) ultimately developed DS and one horse had an FS. Multivariable analysis showed that the severity of sores was positively associated with increasing age (OR: 1.111, P = 0.028), a normal (vs. swollen) limb (OR: 3387, P = 0.023) and an increase in total casting time (OR per week: 1.363, P = 0.002). The thermographic evaluation (35 casts) revealed that the severity of sores was positively associated with increasing deltaT (OR: 2.100, P = 0.0005). The optimal cut-off values for the presence of SS and DS were set at, respectively, deltaT = 23 and 43 degrees C. CONCLUSION AND POTENTIAL RELEVANCE: Distal limb cast is a safe coaptation technique with increasing risk of developing sores with time. Thermography is a valuable and rapid clinical tool to monitor the development of cast sores. PMID- 19301578 TI - Lesions of the deep digital flexor tendon in the digit: a correlative MRI and post mortem study in control and lame horses. AB - REASONS FOR PERFORMING STUDY: Lameness associated with lesions of the deep digital flexor tendon (DDFT) in the digit is now recognised as an important cause of lameness, but there is currently limited information about the pathological nature of the lesions. OBJECTIVES: To compare: signal intensity changes on magnetic resonance images with histopathology; and histopathological changes in the DDFT from horses with no history of foot-related lameness (Group C) and horses with lesions of the DDFT confirmed using magnetic resonance imaging (MRI) (Group D). METHODS: Transverse sections of the DDFT were harvested from 3 sites in all horses: (1) immediately proximal to the navicular bursa (E1); (2) at the level of the navicular bone (E2); and (3) close to the tendon's insertion (E3). If lesions were identified at E1 or had been identified further proximally using MRI, additional sections were obtained until, in most cases, the proximal limit of the lesion was identified. All DDFTs were graded histopathologically using predefined criteria. The MR images were reviewed to determine the location and sequences in which increased signal intensity was seen. RESULTS: No haemorrhage or inflammatory cell infiltration was seen in any horse. At level E1, septal thickening, ghosting of blood vessels and blood vessel occlusion were common in Group D, but were not seen in Group C. Less commonly, there was core necrosis, only seen in Group D. At level E2, septal and vascular changes were most obvious in Group D. Core necrosis, dorsal splitting, crevicing and fibrillation were seen only in Group D. Septal and vascular changes were present in both Groups C and D at level E3, but fibrocartilaginous metaplasia, splitting, crevicing and fibrillation, or core necrosis or fibroplasia were seen only in Group D. Core lesions in Group D often extended proximal to E1, and ranged in length from 0.5 13 cm. Core necrosis was generally associated with increased signal intensity in fat suppressed images. CONCLUSIONS: Lesions of the DDFT in the digit appear to be primarily degenerative, and may be a sequel to vascular compromise. Increased signal intensity on fat suppressed MR images is not necessarily associated with frank fluid or evidence of inflammation, but may reflect major matrix changes in the tendon. POTENTIAL RELEVANCE: Further information about the causes of these lesions is required to develop preventative strategies. PMID- 19301579 TI - Comparison of radiography and ultrasonography to detect osteochondrosis lesions in the tarsocrural joint: a prospective study. AB - REASONS FOR PERFORMING STUDY: Lesions located on the medial malleolus of the tarsocrural joint can be difficult to image radiographically. Ultrasonography allows evaluation of articular cartilage and subchondral bone. OBJECTIVES: To compare dorso30 degrees lateral-plantaromedial-oblique (DL-PIMO) and dorso45 degrees views to detect lesions on the medial malleolus, to validate the use of ultrasonography to show lesions in the tarsocrural joint and to compare its sensitivity to radiography. METHODS: Tarsocrural joints (n=111) with osteochondrosis were evaluated ultrasonographically and radiographically prior to arthroscopic lesion debridement. A complete radiographic examination was made and the best view to detect each lesion recorded. Longitudinal and transverse ultrasonography of the dorsal aspect of the joint was performed and the best scan plane to image each lesion recorded. RESULTS: There were 94 joints with lesions on the distal intermediate ridge of tibia, 24 with lesions on the medial malleolus, and 4 with lesions on the lateral trochlear ridge. The sensitivity of radiography to detect lesions on the medial malleolus and distal intermediate ridge of tibia was 71 and 96%, respectively. Eighty-two percent of lesions on the medial malleolus were better imaged on dorso30 degrees view. The sensitivity of ultrasonography to detect lesions on the medial malleolus and distal intermediate ridge of tibia was 83 and 98%, respectively. Ultrasonography was significantly more sensitive than radiography to detect lesions on the medial malleolus and distal intermediate ridge of tibia. CONCLUSION: Dorso30 degrees lateral plantaromedial-oblique view was the best to image lesions on the medial malleolus. Ultrasonography was a valuable diagnostic tool to diagnose lesions in the tarsocrural joint and was more sensitive than radiography for lesions located on the medial malleolus and distal intermediate ridge of tibia. CLINICAL RELEVANCE: Radiographic examination should include a dorso30 degrees view for detection of lesions on the medial malleolus and ultrasonography should be considered to diagnose osteochondrosis in the tarsocrural joints. PMID- 19301580 TI - Sandwich ELISA system for cartilage oligomeric matrix protein in equine synovial fluid and serum. AB - REASONS FOR PERFORMING STUDY: Measurement of cartilage oligomeric matrix protein (COMP) in serum has potential for diagnosis of equine osteoarthritis (OA), but clinical use is currently limited by the lack of specificity of an inhibition ELISA as well as by baseline increases due to exercise. Improved methods for ELISA with increased antigen specificity and sensitivity are therefore required for reliable measurement. HYPOTHESIS: Measurement of the serum level of COMP by sandwich ELISA allows identification of horses with OA. METHODS: New monoclonal antibodies (mAbs) were elicited against equine cartilage COMP, their epitopes were determined and a sandwich ELISA was developed. The concentrations of COMP in synovial fluid (SF; n=100) and sera (n=100) from OA cases were measured by sandwich ELISA as well as by inhibition ELISA and compared with concentrations in normal joints (n=95) and horses (n=50). RESULTS: Immunoblots of enzymatically cleaved COMP showed that the new mAbs recognised different epitopes located on a 20 kDa fragment between K63 and K238 of the EGF-like repeats. Inhibition ELISA with any mAb detected significantly increased levels of COMP in OA SF compared with normal SF, whereas no significant difference was detected between serum levels of COMP in OA and normal horses. Conversely, sandwich ELISA with the combination of unlabelled 2A11 x biotinylated 11F10 mAbs detected a significant increase in COMP levels in both serum and SF from OA cases compared with levels in normal animals. CONCLUSIONS AND POTENTIAL RELEVANCE: Measurement of serum COMP with sandwich ELISA may be useful in identifying horses with OA. PMID- 19301581 TI - Long-term assessment of horses and ponies post exposure to monensin sodium in commercial feed. AB - REASONS FOR PERFORMING STUDY: Acute monensin intoxication in equids is well described; however, the long-term effects of sublethal intoxication and ability to return to previous use are less well understood. Long-term observations may allow improved estimation of prognosis in cases of sublethal intoxication. OBJECTIVES: To assess horses and ponies exposed to sublethal amounts of monensin for evidence of chronic sequelae and ability to return to prior/intended use. METHODS: Twenty-nine horses and 8 ponies were assessed utilising serum biochemistry, treadmill exercise stress testing, electrocardiography, and pre- and post exercise echocardiography > or = 6 weeks after ingestion of monensin contaminated feed. Animals with evidence of monensin-induced cardiomyopathy were re-examined after a period of rest of > or = 11 months. Follow-up information was obtained by owner telephone interview > or = 52 months after exposure. RESULTS: During resting echocardiography, 11 animals had reduced/low-normal left ventricular fractional shortening (FS); an increase in FS in 8 of these animals was measured > or = 11 months later. Six animals had reduced or low-normal FS during post exercise echocardiography. Two horses had ventricular premature depolarisations during exercise. Follow-up information was available for 35 animals: 21 returned to athletic/reproductive use, 13 were retired immediately and one died. Mean FS increased significantly (P < 0.001) between initial and second examination in 15 animals that underwent resting echocardiography on 2 occasions. CONCLUSIONS: Some equids exposed to sublethal doses of monensin may not develop permanent myocardial disease and a return to athletic/reproductive use is possible. POTENTIAL RELEVANCE: Exercise stress testing, echocardiography and electrocardiography may be useful for detection and monitoring of cardiac dysfunction in equids exposed to monensin and determining whether a return to athletic/reproductive use is possible. PMID- 19301582 TI - Comparison of density gradient and single layer centrifugation of stallion spermatozoa: yield, motility and survival. AB - REASONS FOR PERFORMING STUDY: A new, simpler, technique of colloidal centrifugation has recently been developed, designated single layer centrifugation (SLC). This technique requires evaluation by comparison with a density gradient for its ability to select the best quality spermatozoa and its practicality of use on studfarms. OBJECTIVE: To compare the effect of 2 methods of colloidal centrifugation, density gradient centrifugation and single layer centrifugation, on stallion sperm motility, yield and survival, using freshly collected extended stallion semen. METHODS: Aliquots of extended stallion semen from 10 stallions (38 ejaculates) were processed by the 2 methods of colloidal centrifugation. For both uncentrifuged and centrifuged samples, sperm yield was calculated and subjective sperm motility assessed over several days to provide an estimate of sperm survival. Some stored semen samples, held at 4 degrees C overnight, were also available for testing. RESULTS: For fresh, extended semen, a similar recovery yield of motile spermatozoa was seen for the 2 methods of preparation for single layers and density gradients, respectively. Sperm motility and survival rate were significantly improved by colloidal centrifugation compared to unprocessed ejaculate, without any significant difference between methods (SLC vs. gradient). However, the yield was reduced by 18-20% when cold stored semen was used for centrifugation compared to fresh semen; and more variation between ejaculates was observed than for fresh ejaculates. Again, sperm motility and sperm survival were improved in the centrifuged sperm preparations compared to stored, unprocessed ejaculates. POTENTIAL RELEVANCE: The 2 colloid centrifugation techniques produce equivalent sperm preparations in terms of sperm quality. However, the SLC method would be more practical and convenient for use in the field. PMID- 19301583 TI - Effect of poll flexion and dynamic laryngeal collapse on tracheal pressure in Norwegian Coldblooded Trotter racehorses. AB - REASON FOR PERFORMING STUDY: Dynamic laryngeal collapse (DLC) associated with poll flexion is a newly diagnosed upper respiratory tract obstructive disorder that causes poor racing performance. OBJECTIVES: To determine if Norwegian Coldblooded Trotters (NCTs) affected with DLC associated with poll flexion differ from normal, elite NCTs based on simple airway mechanics measurements. METHODS: Five normal elite NCTs and 6 NCTs diagnosed previously with DLC underwent treadmill videoendoscopy while tracheal pressures were measured continuously. Alternating head positions were used such that horses were exercised with free head carriage and induced poll flexion at heart rates >200 beats/min. RESULTS: Peak inspiratory tracheal pressures were significantly more negative for horses with DLC compared to the elite horses. This difference was only significant during the exercise phases when the poll region was flexed, P = 0.0015. Head position significantly affected peak inspiratory pressure for both elite and affected horses, P < 0.0001. CONCLUSIONS AND CLINICAL RELEVANCE: Induced poll flexion significantly affected peak inspiratory pressure (PIP) in all horses; however, PIPs were significantly more negative in those affected with DLC. Based upon the tracheal pressure measurements recorded in this study, DLC in NCTs is a severe obstructive upper respiratory tract disorder that is induced by poll flexion. PMID- 19301584 TI - Efficacy of thermal cautery for intermittent dorsal displacement of the soft palate as compared to conservatively treated horses: results from 78 treadmill diagnosed horses. AB - REASONS FOR PERFORMING STUDY: Previously, objective comparisons of surgical procedures to relieve dorsal displacement of the soft palate (DDSP) have been limited by the presumptive basis of the diagnostic measures applied. OBJECTIVES: To assess and compare the efficacy of thermal cautery surgery to conservatively treated controls in racehorses definitively diagnosed with idiopathic intermittent DDSP. HYPOTHESIS: Both conservative and surgical treatments have a beneficial result on racing performance in racehorses affected with DDSP. METHODS: Race records were obtained for Thoroughbred racehorses definitively diagnosed with DDSP using high-speed treadmill endoscopy. Racing performance was assessed based on prize money won. RESULTS: Forty-eight horses that underwent thermal cautery and 30 conservatively treated controls were included. Pretreatment earnings significantly decreased in the race immediately prior to diagnosis. A high proportion of previously raced horses returned to racing after both treatments (90-96%). Intrahorse comparison of earnings in 3 races pre- vs. post treatment showed that 53% of conservatively treated horses and 36% of the thermal cautery group had improved performance. Although the difference between these 2 groups may be interpreted as being clinically significant, it was not statistically significant. CONCLUSIONS AND POTENTIAL RELEVANCE: A higher percentage of conservatively treated controls had improved individual performance compared to horses treated with thermal cautery. Thermal cautery appears less effective than other previously published surgical treatments for DDSP. Comparison of the 2 treatment methods should be interpreted cautiously because treatments were not randomised, resulting in baseline variability between groups. PMID- 19301585 TI - Racing performance after combined prosthetic laryngoplasty and ipsilateral ventriculocordectomy or partial arytenoidectomy: 135 Thoroughbred racehorses competing at less than 2400 m (1997-2007). AB - REASONS FOR PERFORMING STUDY: The success of combined prosthetic laryngoplasty with ipsilateral ventriculocordectomy (LPVC) has not been compared to that of partial arytenoidectomy (PA) in a clinical population. HYPOTHESES: In Thoroughbred (TB) racehorses: (1) earnings after LPVC are unaffected by the severity of recurrent laryngeal neuropathy (RLN) (laryngeal grade III vs. grade IV); (2) LPVC and PA yield similar results in the treatment of grade III RLN; (3) performance outcome following PA is independent of diagnosis (RLN vs. unilateral arytenoid chondritis [UAC]); and (4) neither LPVC nor PA returns horses to the level of performance of controls. METHODS: Medical and racing records of 135 TB racehorses undergoing LPVC or PA for the treatment of grade III or IV RLN or UAC were reviewed. Racing records of age and sex matched controls were also reviewed. RESULTS: After LPVC, horses with grade III RLN performed better compared to those with grade IV RLN. Furthermore, horses treated for grade III RLN by LPVC showed post operative earnings comparable to controls. Rate of return to racing were similar for PA and LPVC, although LPVC resulted in higher post operative earnings. Performance after PA was similar regardless of diagnosis (UAC or RLN). Finally, neither LPVC when performed for grade IV RLN, nor PA performed for either diagnosis restored post operative earnings to control levels. CONCLUSIONS: Thoroughbred racehorses treated by LPVC for grade III RLN show significantly better post operative earnings compared to horses treated for grade IV disease. In grade III RLN, LPVC returns earning potential to control levels. PA and LPVC lead to similar success in terms of rate of return to racing, but PA leads to inferior earnings after surgery. POTENTIAL RELEVANCE: Laryngoplasty should be recommended for all TB racehorses with grade III RLN to maximise return to racing at a high level. This contradicts the common approach of waiting for complete paralysis. PMID- 19301586 TI - Expression of toll-like receptor 2 mRNA in bronchial epithelial cells is not induced in RAO-affected horses. AB - REASONS FOR PERFORMING STUDY: Airway inflammation in recurrent airway obstruction (RAO) is triggered by housing affected horses in stables.It has been suggested that RAO is an allergic condition, but innate immune mechanisms are also involved. Fungal products activate innate immune mechanisms through toll-like receptor 2 (TLR2). In human airway epithelium, TLR2 activation leads to interleukin (IL)-8 production. This pathway is negatively regulated by the zinc finger protein A20. This study was performed to enhance understanding of innate immune mechanisms in RAO. HYPOTHESIS: TLR2 and IL-8 mRNA are elevated in RAO during stabling compared with controls. A20 mRNA is negatively associated with the numbers of airway inflammatory cells. OBJECTIVES: To determine TLR2, IL-8 and A20 mRNA expression in lungs of stabled and pastured RAO-affected and control horses. METHODS: Airway obstruction and inflammatory cell counts in bronchoalveolar lavage were measured, and TLR2, IL-8 and A20 mRNA expression quantified by qRT-PCR in 6 RAO-affected and 6 control horses, during and after exposure to hay and straw. RESULTS: Airway obstruction and neutrophils were increased in RAO-affected horses during stabling. While stabling increased IL-8, TLR2 and A20 mRNA were unaffected. TLR2 and A20 were significantly correlated (r = 0.83) and A20 mRNA was negatively associated with inflammatory cells. POTENTIAL RELEVANCE: Stabling does not lead to an increase in TLR2 expression. Other molecules or processes in the TLR2 cascade might be important in fungal-induced airway inflammation. Equine epithelial-derived A20 may be involved in modulation of airway inflammation. PMID- 19301587 TI - Storage-associated artefact in equine muscle biopsy samples. AB - REASONS FOR PERFORMING STUDY: Muscle biopsy is increasingly used in equine veterinary practice for investigating exertional, inflammatory or immune mediated myopathies and unexplained muscle atrophy. Although formalin-fixed samples are often used, for complete evaluation, fresh-frozen tissue is required. Freezing muscle in veterinary practice is impractical: samples sent to specialist laboratories for processing are therefore susceptible to delays, potentially leading to artefact and compromising histological interpretation. HYPOTHESIS: Altered temperature, duration and hydration status influence the severity of storage-induced artefact in equine muscle. METHODS: Skeletal muscle obtained immediately post euthanasia was divided into 6 independent samples from each of 8 horses. One sample per horse was frozen immediately in isopentane precooled in liquid nitrogen. Additional samples were stored in conditions designed to mimic possible situations encountered in practice, including increased storage times, temperature and hydration status. Following storage, stored samples were frozen as before. Cryosections were stained using haematoxylin and eosin and ranked for artefact on 2 occasions by 2 blinded observers. The best samples were processed subsequently with a panel of routine stains and immunolabelled for collagen V to enable the measurement of minimum fibre diameters. RESULTS: Both prolonged storage and increased hydration resulted in more storage-associated artefact. Samples stored for 24 h chilled on dry gauze were ranked higher than those stored on damp gauze; however, a panel of routinely-used histochemical staining techniques was unaffected by chilled 24 h storage. There was no significant effect of storage on mean fibre diameter; however, both chilled dry and damp storage for 24 h caused a significant increase in fibre-size variability. CONCLUSION AND POTENTIAL RELEVANCE: Caution should be exercised when interpreting fibre size profiles in shipped samples. Equine muscle biopsy samples are optimally shipped in dry gauze, sealed in plastic containers and shipped on ice packs to be processed within 24 h and can thus be interpreted by the receiving laboratory with minimal artefact. PMID- 19301589 TI - Rhodococcus equi comes of age. PMID- 19301588 TI - Influenza A viruses with truncated NS1 as modified live virus vaccines: pilot studies of safety and efficacy in horses. AB - REASONS FOR PERFORMING STUDY: Three previously described NS1 mutant equine influenza viruses encoding carboxy-terminally truncated NS1 proteins are impaired in their ability to inhibit type I IFN production in vitro and are replication attenuated, and thus are candidates for use as a modified live influenza virus vaccine in the horse. HYPOTHESIS: One or more of these mutant viruses is safe when administered to horses, and recipient horses when challenged with wild-type influenza have reduced physiological and virological correlates of disease. METHODS: Vaccination and challenge studies were done in horses, with measurement of pyrexia, clinical signs, virus shedding and systemic proinflammatory cytokines. RESULTS: Aerosol or intranasal inoculation of horses with the viruses produced no adverse effects. Seronegative horses inoculated with the NS1-73 and NS1-126 viruses, but not the NS1-99 virus, shed detectable virus and generated significant levels of antibodies. Following challenge with wild-type influenza, horses vaccinated with NS1-126 virus did not develop fever (>38.5 degrees C), had significantly fewer clinical signs of illness and significantly reduced quantities of virus excreted for a shorter duration post challenge compared to unvaccinated controls. Mean levels of proinflammatory cytokines IL-1beta and IL-6 were significantly higher in control animals, and were positively correlated with peak viral shedding and pyrexia on Day +2 post challenge. CONCLUSION AND CLINICAL RELEVANCE: These data suggest that the recombinant NS1 viruses are safe and effective as modified live virus vaccines against equine influenza. This type of reverse genetics-based vaccine can be easily updated by exchanging viral surface antigens to combat the problem of antigenic drift in influenza viruses. PMID- 19301590 TI - [A case of depression with avoidance tendency showing successful reinstatement at work]. AB - Recently, it has been suggested that depressive patients with avoidance tendency are increasing in number. They are inclined to become depressed under specific stressful circumstances, for example, in their workplace. They also show such features as being able to enjoy private activities, to show an almost normal vegetative function, and to blame others or circumstances, etc. Although their symptoms are mild in general, they tend to be prolonged and make it difficult for the patients to return to work. In this article, we report a male middle manager who presented mild depression with avoidance tendency. We tried to reduce his excessive workload, and, at the same time, to keep him going to work. As a result, the rhythm of his daily activities became stable, and work reinstatement was successful. Matussek says that the self has two aspects: the 'private self' and the 'official self'. For depressive patients, the private self is more important, but in public, they tend to do what others want them to. Such behavior with the public self conflicting against the private self makes them exhausted. This is common to the avoidance type of depression, and a typical major depression. The main difference between them is that the orderliness which patients show their behavior, that is, the basic character of the public self, is weak in the former and strong in the latter. In therapy for the avoidance type of depression, it is necessary to both reduce the excessive workload of patients and help them to continually relate to their workplaces, considering the degree of orderliness of their personality. PMID- 19301591 TI - [A nationwide questionnaire survey of medical experts in mental health evaluation]. AB - A nationwide questionnaire survey was conducted to clarify the current status of medical experts in mental health evaluation and their needs (recovery rate: 66.6%). The data were simply tabulated and then statistically analyzed with respect to past experiences of serving as expert witnesses or judges as defined by the Medical Treatment and Supervision Act and past clinical experience. The results showed that 201 respondents (53.0%) had served as medical experts an average of 2.8 +/- 2.4 times, while 173 respondents (45.6%) had not served as such experts, and that 284 respondents (74.9%) had served as judges an average of 3.0 +/- 2.5 times, while 86 respondents (22.7%) had not served as judges. Although about 70% of the respondents who had served as expert witnesses felt that their experiences were burdensome, mostly due to time-related concerns, most had favorable views of the legal process. More than 70% of the respondents indicated that they would continue to serve as expert witnesses despite their sense of burden, thus suggesting a correlation between a past experience of serving as an expert witness and a willingness to do so again in the future. Furthermore, the results indicate that medical experts in mental health evaluation who have not served as expert witnesses are more hesitant, and since physicians employed at institutions for forensic psychiatric examination are more likely to serve as expert witnesses, a relatively small group of medical experts in mental health evaluation repeatedly serve as expert witnesses. On the other hand, when compared to serving as medical experts, serving as judges was psychologically more stressful for many respondents. Since physicians are not used to being judges, they may experience feelings of resistance or burden. Also, judges are somewhat bound by expert testimony, and the fact that there is no written policy for questioning testimony adds to their stress. Therefore, it will be necessary to clarify the positioning of expert evidence and the decisions made by judges. What medical experts in mental health evaluation needed to improve their knowledge varied depending on the type and duration of work and clinical experience. Therefore, it is necessary to provide information matching the experience, skills, and information related to the practical aspect of the Medical Treatment and Supervision Act. Moreover, the results suggest the necessity for medical experts in mental health evaluation to acquire a wide range of expert knowledge in forensic psychiatry beyond that required for the Medical Treatment and Supervision Act. PMID- 19301592 TI - [Intervention and psychological care for the family caregivers of cancer patients]. AB - Recent studies demonstrated that family members of a cancer patient show as much or even higher psychological distress as the cancer patients themselves. There are several reasons for psychological distress among family members. The family of the cancer patient is expected to provide patient care. There are shared responsibilities for decision-making, providing concrete care-giving, meeting the financial and social costs, maintaining stability and adapting to change. In addition, some family caregivers are involved in direct patient care. These responsibilities place both physical and emotional burdens on the family members. It has been shown that spouses demonstrate levels of emotional and functional disruption as great or greater than that of the patient and that these problems often worsen with time, independent of the patient's mood or health. Health problems of caregivers have a very important influence on their ability to meet these demands and it is reported that care-giving affects the physical health of spouse caregivers. Some caregivers may be physically weak or sick, and some may also demonstrate symptoms of cancer. Based on these findings, family members of the cancer patient are called "second-order patients". Therefore, various kinds of care programs are needed to provide support for family members and such interventions are aimed at psychiatric, physical, socio-economic problems. Bereavement is one of the most striking events in human experience. Although bereavement is not a disease, it is associated with excess risk of mortality and morbidity. Bereaved individuals show various physical and psychological symptoms and some of the bereaved individuals develop psychiatric disorders such as bereavement reaction, major depression and post-traumatic stress disorder. Therefore, psychiatric interventions are needed for these bereaved individuals. In the bereavement care clinic, interventions are aimed at psychiatric, physical, socio-economical problems. Among the psychiatric problems, clinicians should consider major depressive disorder because of its higher prevalence, and the need for pharmacotherapy. PMID- 19301593 TI - [Microneurography--from basic aspects to clinical applications and application in space medicine]. AB - Microneurography is an electrophysiological method that directly records impulse traffic from human peripheral nerves by using metal microelectrodes. This method enables the recording of identified postganglionic sympathetic efferent neural traffic leading to the skeletal muscles (muscle sympathetic nerve activity) and skin (skin sympathetic nerve activity), as well as myelinated and unmyelinated afferent nerve impulses from the sensory receptors in skeletal muscles (muscle spindles, Golgi tendon organs, and nociceptors) and skin (skin mechanoreceptors and noci-thermoceptors). The clinical applications of sympathetic microneurography are useful for the elucidation of the neural mechanisms of abnormal blood pressure control and thermoregulation. Sympathetic microneurography is also used in space medicine to elucidate changes in sympathetic neural traffic during and after exposure to simulated microgravity and spaceflight. The clinical applications of sensory microneurography are useful to clarify the neural mechanisms underlying abnormalities in muscle and skin sensation and those in sensory motor control. Microstimulation by using the microneurography technique can also be used to determine the peripheral sensory, sympathetic, and motor nerve functions. PMID- 19301594 TI - [Clinical application of skin sympathetic nerve activity]. AB - Skin sympathetic nerve activity (SSNA) is microneurographically recorded from the skin nerve fascicle in the peripheral nerves. It is characterized by the following features: 1) irregular, pulse asynchronous, burst activity with respiratory variation, 2) burst activity followed by vasoconstriction and/or sweating, 3) elicited by mental stress and arousal stimuli, e.g., sound, pain, electric stimulation, 4) burst with longer duration as compared with sympathetic outflow to muscles, and 5) burst activity following sudden inspiratory action. It comprises vasoconstrictor (VC) and sudomotor(SM) activity, as well as vasodilator (VD) activity. VC and SM discharge independently, whereas VD is the same activity with different neurotransmission. The VC and SM are differentiated by effector response, e.g., laser Doppler flowmetry and skin potential changes. SSNA function in thermoregulation in the human body; however it is also elicited by mental stress. SSNA is the lowest at thermoneutral ambient temperature (approximately 27 degrees C), and is enhanced in the pressence of ambient warm and cool air. The burst amplitude is well-correlated to both skin blood flow reduction rate or sweat rate change. The clinical application of SSNA comprises the following: 1) clarification of sweating phenomenon, 2) clarification and diagnosis of anhidrosis, 3) clarification and diagnosis of hyperhidrosis, 4) clarification of thermoregulatory function and diagnosis of thermoregulatory disorder, 5) clarification of pathophysiology and diagnosis of vascular diseases, e.g., Raynaud and Buerger diseases. 6) clarification of the relation between cognitive function and SSNA and 7) determination of pharmacological effect attributable to change in neuroeffector responses. PMID- 19301595 TI - [Application of microneurography to the evaluation of diabetic neuropathy]. AB - The use of microneurography for recording of the compound nerve action potentials enables a more accurate evaluation of even severe cases of diabetic polyneuropathy. A tungsten microelectrode with a tip diameter of approximately 1 microm is percutaneously inserted into the median nerve trunk in the elbow without the administration of anesthesia. The median nerve in the wrist is subiected to supramaximal electric stimulation, and the largest compound nerve action potential is recorded by the microelectrode. In healthy controls, the median nerve action potential appeared as a large triphasic wave (positive negative-positive; average amplitude, 400 microV) followed by small multiphasic waves. In diabetic patients, the maximal nerve conduction velocity (NCV) and the amplitude of the triphasic wave (Amp) were decreased, and the multiphasic waves were more prominent. The reproducibility of the compound nerve action potentials in the same subject was acceptable. Diminution of Amp appeared to be associated with temporal dispersion due to segmental demyelination. In diabetic polyneuropathy, NCV and Amp decrease with disease progression. In patients with advanced disease and subjective symptoms, low Amp and prominent multiphasic waves were apparent. Because a recording microelectrode is directly inserted into the nerve trunk, this method has many advantages: large potentials measuring several hundred microvolts can be obtained, averaging is not required, and quantitative analysis of the nerve action potential and evaluation of slow fibers are possible. This compound nerve action potential is considered to represent the density of large myelinated fibers (7 microm), especially sensory fibers, around the electrode. This technique may evaluate nerve function more accurately than that of "near-nerve recording of action potentials", a technique reported by Buchthal et al. This method can be a valuable tool for evaluating the function of large myelinated fibers without the use of time-consuming averaging procedures or computer analysis. PMID- 19301596 TI - [Sympathetic neurograms in patients with neurodegenerative disorders--an overview]. AB - Sympathetic nerve functions in patients with amyotrophic lateral sclerosis (ALS), spinocerebellar degeneration (SCD), and Parkinson disease (PD) were evaluated using microneurography. In patients with ALS, the muscle sympathetic nerve activity (MSNA) at rest was greater than that in healthy subjects and patients with other neuromuscular disorders. This finding is not observed in the advanced stages of ALS. The resting frequency of skin sympathetic nerve activity (SSNA) significantly greater in ALS patients than in healthy controls. In patients with SCD, the MSNA was slightly reduced, although these patients did not experience orthostatic dizziness or syncope, or significant fall in blood pressure during the head-up tilt test. The reflex latency of SSNA induced by electric stimulation was slightly but significantly prolonged in patients with cortical cerebellar atrophy and markedly prolonged in patients with multiple system atrophy-C (MSA C). In patients with PD, a significantly negative correlation was observed between MSNA and age, and between MSNA and disease duration. In these patients, the resting frequency of SSNA was significantly lower than in healthy controls, while SSNA reflex latencies were similar to those in the controls. Further investigations to determine whether the autonomic nervous system was impaired are imperative for elucidating the pathogenesis of neurodegenerative disorders. PMID- 19301597 TI - [Application of microneurography to circulatory disorders]. AB - Sympathetic neural function has been conventionally evaluated by muscle sympathetic nerve activity (MSNA), plasma norepinephrine levels, and heart rate variability. Of these, MSNA is the only direct method for the assessment of sympathetic neural outflow to the peripheral vasculature. However, interindividual comparison of MSNA is limited to burst counting. Recently, we proposed a new quantitative and objective measure of MSNA--burst power--which enables to assess inter-individual differences in MSNA. The sympathetic nervous system regulates circulation via arterial and low-pressure baroreflexes, lung stretch reflex, chemoreflexes, and other somatic and sympathetic afferents. The open-loop gain of the arterial baroreflex system can be determined on the basis of the transfer function from the random input of arterial blood pressure to the output of MSNA using the autoregressive moving average model with an exogenous input (ARMAX). This model revealed that baroreflex-mediated sympathetic control was preserved while the reflex parasympathetic control was markedly attenuated in patients with heart failure. The lung stretch reflex, simulated by the ARMAX using the input of instantaneous ventilation and the output of MSNA, showed that instantaneous lung inflation suppressed MSNA within 1.5 sec. The magnitude or the lung stretch reflex decreased as the heart failure progressed. In patients with heart failure, the carbon dioxide chemoreflex is enhanced, as evidenced by substantial increases in both ventilation and MSNA on exposure to carbon dioxide. Thus, the use of new signal processing techniques and simultaneous assessment of MSNA and other hemodynamic and respiratory variables could provide a new insight into the dynamic function of cardiovascular and respiratory neural reflexes. PMID- 19301598 TI - [Additional further applications of microneurography in the future]. AB - Microneurography has been mainly used in the field of basic science for obtaining information on the physiological phenomena of normal volunteers. Some clinical studies have been reported in the field of the autonomic nervous system. In the future, further applications of microneurography in the clinical situations will be desired. One of such situations is emergency or acute phase of cerebrovascular diseases, for example, at bedside or in intensive care units. Further, the recording equipment would be desired. A portable device was newly developed and is now available for this purpose of information acquistion. Another application of microneurography, i. e. as a research technique in a field intermediate between medical and engineering was introduced. The project aimed to develop an artificial hand that could sense various external stimuli and permit voluntary finger movement. Microneurography was used for conducting the impulses from the surface pressure transducer on the artificial hand and triggering pressure sense in the subject via microstimulation of inserted nerve fibers. Great attention should be paid to insert the needle into various tissues other than a nerve. The epoxy resin coating for insulation was stripped irregularly with the aid of observation using scanning electron microscope. The optimum stripping method for recording nerve activity when tungsten microelectrode needles are employed is still unknown. The insertion technique can be analyzed by ultrasonography. The identification of the needle on the ultrasonographic image is often difficult; however, the needle direction can be easily judged by observation of the motion of the surrounding tissue. PMID- 19301599 TI - [TRIAD--a form of type 3 cell death]. AB - Functional disturbances of neurons are implicated in neurodegenerative disorders including Alzheimer's diseases and polyglutamine diseases, while the connection between neuronal dysfunctions and cell death remains unclear. In this review, transcriptional repression-induced atypical cell death of neurons (TRIAD), a form of type 3 cell death, is reviewed in connections with surrounding knowledge. PMID- 19301600 TI - [New approach to apraxia]. AB - Our collective understanding of our approach to apraxia has not dramatically changed over the last century. The classical approach to apraxia has occasionally caused confusion and disagreements because it lacks established definitions for apraxia and related terms such as "praxicon" or "ideation". Rather, the current system allows each researcher to define these terms differently, making it extremely difficult to compare cases reported by different researchers. New approaches, however, are providing more concrete definitions, enabling us to describe apraxic disorders more operationally, this may help prevent the disagreement concerning these definitions. A more recent approach, demonstrated in several reports, focuses on the interactions between "body" and "objects". Another approach concentrates on information processing such as "online processing" and the retrieval of stored knowledge. These new approaches to apraxia have the potential to improve our ability to communicate about this condition. In future studies, these conceps should be integrated into the classical approach to create a single, workable model for understanding apraxic syndromes. PMID- 19301601 TI - [Study of cumulative head injury in high shock avoider rat (THA rat)]. AB - PURPOSE: Small but repeated head trauma, as represented by boxing-related punch drunk syndrome and dementia pugilistica, occasionally cause dyskinesia and marked brain dysfunction following long-term post-traumatic follow-up, despite the absence of intracranial lesions, such as cerebral contusion and intracranial hemorrhage. We defined this condition as "cumulative head injury." To clarify its mechanism, we conducted an experiment involving appliciation of continuous head trauma of Tokai High Avoider (THA) rats, and examined subsequent marked function/histopathological changes. METHODS: THA rats were divided into 3 categories based on the frequency of impact exposure: a control group (Group A), a group exposed to 1 impact set (Group B), and a group exposed to 3 impact sets (Group C). In each group, histopathological, spontaneous motility, and learning tests were conducted. RESULTS: Histopathologically, no marked tissue destruction was observed in Group B or C. In Group C, the number of GFAP-positive cells were increased in acute-phase specimens of the hippocampus, cerebral cortex, and basilar cortex. With respect to chronic-phase histological changes, the numbers of GFAP-positive cells were increased in the hippocampus and the basilar cortex in Group C; however, these changes were less marked than in the acute stage. A marked function test identified emotional suppression in the acute stage and bimodal learning reduction in the acute to chronic stages in Group C. CONCLUSION: The results of this experiment revealed that the repetition of low-level trauma which did not lead to brain injury as revealed on pathological examination, induced emotional suppression and the bimodal reduction in learning results; further, this disorder exacerbated with an increase in impact frequency. The influence on marked brain function could be verified using a specific experimental system of THA rats. This model may be useful for evaluating the cumulative effects of repeated head trauma. PMID- 19301602 TI - [Case of Leber's hereditary optic neuropathy with mitochondrial DNA 11778 mutation exhibiting cerebellar ataxia, dilated cardiomyopathy and peripheral neuropathy]. AB - We report the case of a 28-year-old woman with Leber's hereditary optic neuropathy (LHON) associated with cerebellar ataxia, dilated cardiomyopathy and peripheral neuropathy. She had a mitochondrial DNA point mutation from guanine to adenine at nucleotide position 11778 and developed ataxic gait within 2 years after the onset of bilateral visual loss. A neurological examination detected horizontal nystagmus, bradylalia, and truncal and bilateral limb ataxia of the cerebellar type. She could walk, albeit unsteadily. There was no weakness in her arms and legs. Tendon jerks were diminished in both the upper arms. Bilateral knee and ankle jerks were absent, and the plantar responses were neutral. Paresthesia of the stocking type was present but no reduction of pinprick, position or vibration senses was detected in the paresthetic regions. Romberg's sign was negative. Brain MRI showed atrophic changes in both the cerebellar vermis and the hemispheres. Nerve conduction studies detected mildly decreased motor nerve conduction velocities in the median, ulnar and posterior tibial nerves. Ultrasound cardiography showed a dilated left ventricle. It was not possible to clarify the relationship between LHON and cerebellar atrophy, cardiomyopathy, and peripheral neuropathy. However, physicians, need to be aware that the patients may develop various neurological complications after the onset of optic neuropathy in LHON. PMID- 19301603 TI - [Case report of intrathecal baclofen administration in a child with severe spasticity attributable to traumatic brain injury]. AB - We report a pediatric case of intrathecal baclofen therapy for severe spasticity following traumatic brain injury. A 14-year-old boy suffered from traumatic brain injury by traffic accident in 2005. Subsequently, he experienced tetraparesis and severe spasticity with spontaneous spasms. He underwent baclofen screening test, and his spasticity was improved. Thereafter intrathecal baclofen therapy was performed. Following baclofen pump implantation, Ashworth Score decreased from 4.0 points to 3.0 in lower limbs, and from 3.0 to 1.5 in upper limbs. His muscle tone was reduced and occurrence of spontaneous spasms stopped. Intrathecal baclofen therapy was observed to be an effective treatment for severe spasticity in childhood. Since children receiving the therapy demonstrated longer survival period than adults, long-term follow-up of this therapy is warranted. PMID- 19301604 TI - [Case of infected subdural hematoma diagnosed by diffusion-weighted imaging]. AB - An 80-year-old man who had undergone total gastrectomy and splenectomy for gastric cancer 13 years ago presented with headache, drowsiness, and high fever 1 month after a traffic accident. Brain CT scans revealed bilateral subdural fluid collections. Diffusion-weighted imaging (DWI) showed mixed high and low signal intensities in the left subdural fluid, and contrast-enhanced MR imaging revealed capsule enhancement of the left subdural fluid collection. The patient was diagnosed with left subdural empyema, and 2 burr-holes were drilled for drainage and irrigation. Operative findings revealed a neomembrane underneath the dura mater. Old hematoma and yellowish-white purulent fluid were present within the neomembrane. This confirmed the diagnosis of infected subdural hematoma (ISH). Abscess culture results were positive for Escherichia coli. The patient's symptoms resolved postoperatively with subsequent antibiotic therapy. However, 4 months after the operation, he suddenly died of severe sepsis and disseminated intravascular coagulation following cholecystitis, which was possibly associated with splenectomy. The clinical presentation, diagnosis, and treatment of an unusual case of ISH have been discussed. We emphasize that DWI and enhanced MR imaging may be useful for diagnosing ISH, and serial DWI evaluations may help in monitoring the therapeutic response in ISH. PMID- 19301605 TI - [Posttraumatic pseudomeningocele--a case report]. AB - Pseudomeningocele is a rare complication of head trauma. An 7-year-old male child who fell from playground equipment was admitted to hospital with lambdoid sutural diastatic fracture. As epidural and subgaleal fluid collections gradually developed from the subsequent day of admission, a subgaleal drain was placed for cerebrospinal fluid (CSF) diversion. In addition, the scalp just over the fractured segment was compressed by band after drainage whereby the CSF leak was stopped. On the basis of our experience with this case, we suggest that appropriate management enables repair of posttraumatic pseudomeningocele in early stages of injury. PMID- 19301606 TI - [Large optic nerve sheath meningioma]. PMID- 19301607 TI - [One hundred books which built up neurology (27)--Marshall Hall "synopsis of the diastaltic nervous system" (1850)]. PMID- 19301608 TI - [Clinical studies of renal pelvic and ureteral cancers--comparison with or without asymptomatic macrohematuria at initial complaint]. AB - We analyzed 95 patients with renal pelvic and ureteral cancers with total nephroureterectomy at our Department fromJanuary, 1997 to April, 2007. The patients were composed of 60 males and 35 females, between 41- and 90-years-old with a median age of 69-years-old. Histologically, there were 94 transitional cell carcinomas (TCC) and 1 TCC with squamous cell carcinoma. Fifty five cases (57.9%) were less than pT2, and 41 cases (42.1%) were more than pT3. Findings of lymphatic and venous invasion were present in 35 cases and 24 cases respectively. The overall survival rate at 5 years was 76.6%. In this series, pathological stage, infiltration pattern, lymphatic and/or venous invasion and the regional lymph nodes indicated a significantly poor prognosis. On the other hand, multivariate analysis using Cox proportional hazards regression revealed the presence of pathological stage as the most significant predictor of survival. Furthermore, we investigated histopathological prognostic factors of the patients separating asymptomatic macrohematuria group (AM group) and non-asymptomatic macrohematuria group (non-AM group). According to this analysis, non-AM group had worse pathological stage, grade and lymphatic invasion than AM group. PMID- 19301609 TI - [A clinical study concerning survival of operative cases for the transitional cell carcinoma of the upper urinary tract]. AB - To identify prognostic factors influencing survival in transitional cell carcinoma (TCC) of the upper urinary tract, we retrospectively studied 189 (median age 70, 130 males and 59 females) patients who underwent surgical resection at our 3 hospitals from August 1980 to September 2006. After a median follow up of 47.8 months, 45 cases (23.8%) died of cancer and the 5-year and 10 year disease-specific survival rate (Kaplan-Meier method) was 70.5 and 67.1%. Because lymphatic and vascular involvements, pattern of infiltration and location of tumor had loss of data, only the univariate analysis was done. They were observed to be significantly different by the univariate analysis. The significant prognostic factors for survival using Cox-proportional hazard models were tumor stage, tumor grade, lymph node metastasis, and surgical margin status. Adjuvant chemotherapy was not the prognostic factor in our multivariate analysis. PMID- 19301610 TI - [Influence of blood cells in urine samples on results of screening for urothelial carcinoma with NMP22 bladder chek]. AB - We compared the sensitivity of Bladder Chek NMP22 with that of urine cytology in bladder cancer patients. Further, we evaluated the usefulness of Bladder Chek NMP22 in patients with benign diseases such as cystitis, urolithiasis, and benign prostate hyperplasia (BPH) and examined how blood cells in urine samples affect the results of Bladder Chek NMP22. Patients with macroscopic hematuria were excluded from this study. Of 77 bladder cancer patients, Bladder Chek NMP22 showed positive in 46.8%, while urine cytology in 33.8% (p = 0.16). Bladder Chek NMP22 and urine cytology showed positive in 31.8 and 0.0% in G1 (p = 0.004), 51.2 and 46.3% in G2 (p = 0.66) and 57.1 and 50% in G3 (p = 0.71); 44.4 and 88.9% in Tis (p = 0.052), 25.6 and 15.4% in Ta (p = 0.27), 72.2 and 33.3% in T1 (p = 0.02) and 81.8 and 54.5% in T2 or higher (p = 0.18), respectively. In bladder cancer patients with microscopic hematuria or pyuria, the positive rates of Bladder Chek NMP22 were 82.1 and 73.1%, respectively, whereas they were 26.5% (p < 0.001) and 33.3% (p = 0.002), respectively, in those without hematuria or pyuria. In 36 cystitis, 20 urolithiasis, and 19 BPH patients, the positive rates of Bladder Chek NMP22 were 58.3, 25.0 and 5.5%, respectively. Bladder Chek NMP22 showed higher sensitivity for detection of bladder cancer, especially in low-grade and low-stage cancers than urine cytology, but the result was likely affected by blood cells in urine samples. Thus, although Bladder Chek NMP22 may be less useful as the first device for screening of urothelial cancer in patients with hematuria or pyuria, it may show results of high quality when used in patients with negative urine cytology after excluding benign diseases. PMID- 19301611 TI - [Risk factors for surgical site infection in radical cystectomy: efficacy of buried suture by absorbable thread for skin closure]. AB - We assessed the incidence, risk factors and causative organism for surgical site infection (SSI). We reviewed the clinical charts of the consecutive 100 patients who underwent radical cystectomy from December 2003 to April 2008. All patients received intravenous prophylactic antimicrobial agent, 1 gram of cefotium hydrochloride before surgery followed by the same agent twice a day through postoperative day 3. Skin closures were carried out by interrupted sutures employing a 3-0 nylon for the first 50 patients (interrupted group), and by buried suture employing 3-0 or 4-0 braided polyglactin for the last 50 patients (buried group). SSI occurred in 13 cases (26%) with interrupted group including 8 patients with ureterocutaneoustomy and 5 patients with orthotopic ileal neobladder reconstruction. On the other hand, SSI was found in 1 patient (2%) in the buried group for whom ileal neobladder reconstruction was carried out (P= 0.021). Superficial inflectional SSI was found in 6 patients (5 patients in the interrupted group, patient in the buried group) and deep incisional SSI occurred in 5 patients in the interrupted group only. The most frequently isolated organism was Enterococcus faecalis. Univariate analysis revealed that body mass index (BMI) and the skin closure methods were significant risk factors for SSI (P<0.001, P=0.021, respectively). These 2 risk factors for SSI were also significant in multivariate analysis (P=0.003, P=0.037, respectively). These results suggested that BMI and the skin closure methods are independent risk factors in radical cystectomy. Furthermore, it may be possible to reduce the incidence of SSI by performing the buried suture for the skin closure. PMID- 19301612 TI - [Histopathologic changes due to combination of androgen deprivation and transurethral microwave thermotherapy in patients with localized prostate cancer]. AB - Fifty patients with localized prostate cancer, clinical stage T1-T2N0M0 were studied. The therapy started with androgen deprivation therapy for 3 months to reduce the volume of prostate about 35%. High-energy transurethral microwave thermotherapy (TUMT) was then performed. Transurethral resection of the prostate (TURP) was performed in radical fashion at least 3 months after TUMT to confirm the treatment effect. Thorough pathological study of all the TURP chips revealed no cancer cells in 39 of the 50 patients. Among 11 patients who revealed remnant of cancer cells, 4 patients were with viable cancer cells, and 7 were with devitalized cancer cells. PMID- 19301613 TI - [Analysis of the statues of patients visiting our specialized clinic for hypogonadal men: 5-year experience]. AB - A specialized clinic for middle-aged and elderly hypogonadal men was established in our institution five years ago. A retrospective study of the 511 patients who attended our clinic during this period was conducted and the issues involved in treating these patients were identified. The patients' age distribution, symptoms, serum testosterone values, treatment, and course were examined. The patients' mean age was 54.0 years (range, 35 to 74 years); approximately half of the patients were in their fifties. Most patients complained of decline in libido, general fatigue, and erectile dysfunction. The patients' mean serum free testosterone was 9.28 pg/ml; 40.3% of patients had a serum free testosterone value of less than 8.5 pg/ml, which is the threshold value for the initiation of androgen replacement therapy (ART) in Japan. ART was given to 220 (43.1%) patients, and it was considered effective in 100. It appears that male climacteric disorder and late-onset hypogonadism have become commonly recognized in Japan. There will be an increasing need for specialized clinics for such men in the future. PMID- 19301614 TI - [Four cases of sarcomatoid renal cell carcinoma]. AB - We herein report four cases of sarcomatoid renal cell carcinoma. These cases comprised 4.1% of the 98 patients with renal cell carcinoma treated in our department during the past 13 years. It is confirmed that renal cell carcinoma with a sarcomatoid component often shows local invasion, distant metastasis, rapid growth and poor prognosis. In Mian's series, the percentage of sarcomatoid component (< 25% vs > or = 25%) was associated with decreased survival time, but was independent of stage. The pathological stage was pT3b N0 M0 in case 1, pT1b N0 M1 in case 2, pT3b N0 M1 in case 3 and pT1a N0 M1 in case 4. The pT1 sarcomatoid renal cell carcinoma in case 2 and case 4, who developed poor prognosis, was composed of 60 and 80% sarcomatoid change, respectively. However in case 3 with a pathological stage of pT3, the patient is alive 35 months after resection, because the extent of sarcomatoid component was 25%. The prognosis of patients with sarcomatoid renal cell carcinoma depends on not only disease stage and tumor grade but also the pathological extent of sarcomatoid component. PMID- 19301615 TI - [A case report of renal cell carcinoma with metastatic intraductal tumor thrombus of the common bile duct]. AB - We present a case of liver metastasis of renal cell carcinoma (RCC) presenting as an intraductal tumor thrombus of the common bile duct. A 62-year-old man was admitted with jaundice. He had undergone right nephrectomy for RCC 8 years previously, and had been administered interferon alfa for lung, bone, and liver metastases for these 3 years. Abdominal enhanced computerized tomography (CT) disclosed the progression of liver metastasis to the common bile duct and the dilated bilateral bile ducts. Endoscopic retrograde cholangiopancreatography (ERCP) revealed a smooth filling defect in an upper portion of the common bile duct, suggesting an intraductal RCC tumor thrombus of the common bile duct. He died 4 months later despite the percutaneous biliary drainage. Liver metastasis of RCC proceeding to the common bile duct is extremely rare, and this is the 8th case. We should be aware of the fact that metastatic intraductal tumor thrombus of the common bile duct can cause obstructive jaundice. PMID- 19301616 TI - [Testicular tumor detected in a male infertility patient with prior orchiopexy]. AB - A 37-year-old man was referred to our hospital with the chief complaint of infertility. Semen analysis at another clinic had demonstrated severe oligozoospermia and he had a history of bilateral orchiopexy for undescended testes. Scrotal ultrasonography revealed a hypoechoic heterogeneous mass (1.5 cm in diameter) within the left testis. With a provisional diagnosis of testicular tumor, left high orchiectomy was performed. Pathological examination of the tumor revealed seminoma. At 12 months after surgery, there has been no evidence of recurrence. Clinicians should be aware of the link between male infertility and testicular cancer, particularly following orchiopexy. PMID- 19301617 TI - [Male hypogonadotropic hypogonadism (MHH) outpatient with testosterone deficiency syndrome: a case report]. AB - A 59-year-old male was referred to our hospital with chief complaints of general fatigue and muscle stiffness of the shoulders. His hormonal data were total testosterone 0.05 ng/ml (normal: 2.01-7.5 ng/ml), free testosterone less than 0.6 pg/ml, leuteinizing hormone (LH) 1.1 mIU/ml (2.2-8.4 mIU/ml), follicle stimulating hormone (FSH) 2.4 mIU/ml (1.8-12 mIU/ml) and prolactin (PRL) 13.1 ng/ml (4.3-13.7 ng/ml). Though both his genital stage and pubic hair stage were Tanner V, his testis volume was 12 ml on the right and 10 ml on the left. A gonadotropin releasing hormone (GnRH) stimulation test revealed low responses of LH and FSH. Magnetic resonance imaging of the head revealed pituitary tumor (15 mm). Our diagnosis was acquired hypogonadtropic-hypogonadism-related pituitary tumor. Transsphenoidal hypophysectomy was performed and pathological diagnosis revealed epidermoid cyst. Three months after the operation, his total testosterone was elevated to 1.17 ng/ml and his complaints were improved. PMID- 19301618 TI - Who should speak at the people's house? PMID- 19301619 TI - Ubiquitin-fusion protein system: a powerful tool for ectopic protein expression in mammalian cells. AB - Clonal cell lines stably expressing a protein of interest are frequently used in the characterization of the potential physiological functions of mammalian proteins, but the generation of such cell lines is laborious and time-consuming. Here, we made use of the fact that fusion proteins consisting of ubiquitin linked to the N terminus of a protein of interest are efficiently processed by ubiquitin specific proteases into their respective free proteins within eukaryotic cells, and designed an expression system that allows easy and highly efficient selection of mammalian cells ectopically expressing a protein of interest. PMID- 19301620 TI - Dispensing of very low volumes of ultra high viscosity alginate gels: a new tool for encapsulation of adherent cells and rapid prototyping of scaffolds and implants. AB - We present a tool for dispensing very low volumes (20 nL or more) of ultra high viscosity (UHV) medical-grade alginate hydrogels. It uses a modified piezo-driven micrometering valve, integrated into a versatile system that allows fast prototyping of encapsulation procedures and scaffold production. Valves show excellent dispensing properties for UHV alginate in concentrations of 0.4% and 0.7% and also for aqueous liquids. An optimized process flow provides excellent handling of biological samples under sterile conditions. This technique allows the encapsulation of adherent cells and structuring of substrates for biotechnology and regenerative medicine. A variety of cell lines showed at least 70% viability after encapsulation (including cell lines that are relevant in regenerative medicine like Hep G2), and time-lapse analysis revealed cells proliferating and showing limited motility under alginate spots. Cells show metabolic activity, gene product expression, and physiological function. Encapsulated cells have contact with the substrate and can exchange metabolites while being isolated from macromolecules in the environment. Contactless dispensing allows structuring of substrates with alginate, isolation and transfer of cell-alginate complexes, and the dispensing of biological active hydrogels like extracellular matrix-derived gels. PMID- 19301621 TI - Conditional protein stabilization via the small molecules Shld-1 and rapamycin increases the signal-to-noise ratio with tet-inducible gene expression. AB - Cellular mechanisms control one or more of the three basic levels of regulation (transcription, translation, and protein activity/locality), allowing for finely tuned spatial and temporal regulation of protein expression patterns. Gene regulation constructs in wide use today often employ a constitutively expressed transcription factor whose activity is determined by the presence or absence of a small molecule. A case in point is the tet transcription system, wherein transcription is regulated by doxycycline (Dox), allowing the researcher to turn protein expression on or off depending on the presence/absence of Dox. However in many applications of that system, there is basal transcription from the promoter element that is independent of Dox. Moreover, in vivo, heterogeneous distribution of Dox leads to concurrent differences in gene expression. We addressed these limitations by introducing conditional destabilizing elements to the system. First, we created a transactivator protein fusion regulated at the additional level of protein stability. This modification enabled a system that demonstrated an off state that is less sensitive to variations in Dox concentrations. We also regulated the stability of the protein expressed from the tet operator cassette, observing greatly improved signal-to-noise ratios. The results underscore how investigator-defined regulation at multiple levels of protein expression can attain afiner degree of control over the final expression of introduced genes. PMID- 19301622 TI - Optimization of 454 sequencing library preparation from small amounts of DNA permits sequence determination of both DNA strands. AB - To increase the yield of DNA sequence generated by the 454 technology from small amounts of starting DNA, we investigated the efficiency of each step in the 454 library preparation process. We find that the last step, when the single-stranded library is released by NaOH, is inefficient and highly variable. When this step is replaced with heat treatment, library amounts dramatically increase. Furthermore, when sequencing templates are first isolated by NaOH treatment and subsequently by heat treatment, the sequences of both strands of individual template DNA molecules can be determined. Using this approach, we confirm that C/G base pairs observed as T/A base pairs in Neanderthal DNA sequences are due to a modification of the cytosine rather than guanine residues. PMID- 19301623 TI - Genotyping of IRGM tetranucleotide promoter oligorepeats by fluorescence resonance energy transfer. AB - Fluorescence resonance energy transfer (FRET) genotyping has been well established for the rapid assessment of single nucleotide polymorphisms (SNPs) and deletions. A design is presented that allows the typing of short tandem oligo repeat sequences using the LightTyper/LightCycler system. The protocol was evaluated and applied to the typing of a tetranucleotide promoter repeat of the human gene encoding the immunity-related GTPase family, M (IRGM) molecule in >2000 individuals from Ghana, West Africa. PMID- 19301624 TI - Comparing the clinical effect of five varying locations of LI.4 acupoint. AB - LI.4 is a major acupoint but the method of locating it has not been standardized. In fact, description of method for locating this acupoint often varies in the classic and traditional texts. It might signify this point may be varied from one to another person. Our comparative study of locating and subsequent acupuncturing these locations revealed some interesting features of LI.4 from our collected clinical data in that location 1 had a better therapeutic effect for toothache relief and analgesia-anesthesia effect than for headache relief, location 2 was better for headache than toothache relief; locations 3 and 4 had a mixed effect whereas, location 5 seems to have a better chance to get bioenergy (De Qi) as manifested by tingling and numbing sensation at the acupunctured site. Our study also suggested that LI.4 location was about the size of 4 millimeters in diameter instead of a pin-point location, though needle inserted at correct point produced a better result. PMID- 19301625 TI - The antipruritic effect of acupuncture on serotonin-evoked itch in rats. AB - The antipruritic effect of acupuncture was studied using a rat model of hindlimb scratching. After acupuncture or electroacupuncture (EA), which was conducted for 30 min, itch-associated behavior was induced by an intradermal injection of 2% serotonin (20 microl) into the rostral back, and then numbers of scratching bouts were counted for 60 min. During the first experiment, acupuncture stimulations were applied to several different points. However acupuncture significantly reduced numbers of scratchings only when applied to cervical dermatomes. In the second experiment, plain acupuncture, or 2Hz, or 120Hz EA were applied to acupoints LI 11 and LI 4, at which acupuncture stimulation produced the greatest antipruritic effect in the 1st experiment, and as serotonin was administered in the same manner described for the 1st experiment. Results showed that 2Hz EA stimulation tended to increase pruritic bouts by approximately 18% versus the animals treated with plain acupuncture, whereas 120Hz EA stimulation tended to decrease pruritic bouts by approximately 39% compared with animals subjected to plain acupuncture. When nor-binaltorphimine (a kappa-opioid receptor antagonist) was pretreated to elucidate the relation between kappa-opioid receptor and the antipruritic effect of 120Hz EA, it was found to markedly inhibit the antipruritic effect of 120Hz EA. These results suggest that acupuncture and EA stimulation are effective treatments for pruritus if administered to dermatomes corresponding to affected sites or to adjacent dermatomes and that this effect is due to the antipruritic effect of kappa-opioid receptor activation maximally induced by high-frequency EA stimulation. PMID- 19301626 TI - The role of the intensity of low frequency electroacupuncture stimulation on the modulation of capsaicin-induced edema in the rat paw. A blind controlled study. AB - Experimental animal and clinical human data suggest that electroacupuncture (EAP) reduces the release of substance P from sensitive neurons, both at medullar level and in the periphery. Aim of our study is to verify the effect of different intensities of stimulation on edema induced by subcutaneous administration of capsaicin. The study was performed on 72 male Sprague-Dawley rats divided into 4 experimental groups according to the intensity of electrostimulation (5, 10, 50, 70 mA) and a control group. A constant current electrical stimulator delivering positive and negative biphasic wave (duration of one pulse wave complex: 500 microsec; pulse repetition rate: 5/sec) has been used for the stimulation. The lowest intensity of stimulation (5 mA) was effective in the prevention of neurogenic edema. Conversely, higher stimulation intensities, namely 10 and 50 mA, were not effective in reducing edema. Stimulation at 70 mA caused a worsening of edema, probably due to an increased release of substance P in the paw. PMID- 19301627 TI - Heat transfer involved in a warm (moxa-heated) needle treatment. AB - A numerical study was performed to reveal the temperature distributions in tissues around the GB38 acupoint during the warm (moxa-heated) needle treatment (WNT). The proposed WNT model involves a needle, needle handle and the calf section around the GB38 acupoint. For validating the numerical results, measurement using the calibrated infrared (IR) camera was also conducted. Our aim was to calculate the skin surface temperature and the temperatures under warm needle at different depths beneath the skin. From the temperature profile along the centerline of the acupuncture needle, the temperature is seen to decrease rapidly from the burning moxa section (200 degrees C) to the skin surface (36 degrees C). The temperature near the needle (36 degrees C) is only 1 degrees C higher than those predicted at other places (approximately 35 degrees C). This phenomenon explains why this treatment practice is called the warm needle rather than the hot needle treatment. PMID- 19301628 TI - Randomized control trial of hand acupuncture for female stress urinary incontinence. AB - This study was designed to evaluate the effect of Hand acupuncture treatment on the stress urinary incontinence of 52 women randomly assigned to Experimental hand acupuncture treatment group (n = 25) and Control group (n = 27). 11 hand acupuncture points were established as a basic treatment formula. Among them, 5 points on the midline of the palm represent the major acupoints of external genitals, bladder, abdominal aorta, stomach and heart, while the other 6 points were applied as supplementary ones (2 points on the mid line of the palm for adrenal glands and heart, 2 points near the wrist for kidneys and 2 points on the back of the hand for bladder). This basic treatment formula was applied to all subjects in the Experimental group. Additional treatments were given to the subjects with tenderness at ST27, CV4 or SP15, while no treatment to the subjects in Control group for 12 weeks. Assessment was made before and after 12 weeks of clinical study. In the present study, Hand acupuncture treatment significantly reduced the frequency of urinary incontinence compared to Control group. Similarly, the prevalence score of urinary incontinence was significantly reduced up to 37% of Control group in Experimental group. Furthermore, vaginal contraction pressure of the Experimental group was significantly increased 1.6 fold compared to Control group. In addition, Hand acupuncture treatment significantly improved the scores of making friends, sexual life and social community in Experimental group compared to Control group. Overall, these findings suggest that Hand acupuncture can be a potent alternative therapy for stress urinary incontinence. PMID- 19301629 TI - Organ representation areas of different internal organs localized on the eyebrows & eyelids by the Bi-Digital O-Ring Test electromagnetic resonance phenomenon between 2 identical molecules--Part I: Detection of increased markers for cancer & heart disease from white hairs on the eyebrows and marked decrease in the markers after taking one optimal dose of DHEA. AB - Organ representation areas of different internal organs were localized on the eyebrows for the first time in 2005 and localized on the surrounding tissue around the eye in 2008 using the Bi-Digital O-Ring Test (BDORT) electromagnetic resonance phenomenon between 2 identical tissue. The appearance of white hairs at specific organ representation areas of eyebrows represent an abnormal condition of the specific internal organ if the BDORT grading is (-)4 or a higher negative value. When one of these white hairs on the eyebrows is examined at any location along its entire length by the BDORT, if the grading is a very abnormal negative value of (-)6 or a higher (-) value, significantly abnormal conditions exist or existed at the corresponding time at the specific location of the hair. The heart representation area on the eyebrow is located nearest to the bridge of the nose on each side, where abnormally increased BNP (Brain Natriuretic Peptide) and cardiac Troponin I content is detected on the part of the white hair where the BDORT grading is (-)6 or a higher negative value. In the very abnormal part of the white hair representing the heart, Asbestos, TXB2, and PLGF (Placental Growth Factor) are markedly increased, while DHEA is markedly reduced. If Integrin alpha5beta1 and Oncogene C-fos Ab2 are more than 5-10 ng (BDORT units), malignancy can be suspected. When one optimal dose of DHEA, 8.5 mg, was taken once, normal body tissue telomere was increased from less than 1 ng (during which time the patient experienced extreme tiredness and frequent urination), to 525 ng (BDORT units), not only did severe tiredness & frequent urination completely disappear, but also the white hair representing the left ventricle changed to black hair, and the semi-transparent white hair representing the kidney changed to a non-transparent white hair, with significant decreases in the abnormal parameters. Thus, it is possible to detect unrecognized serious heart conditions or early-stage cancer that are currently present or that were present in the recent past by examining the white hairs of the eyebrows, and these diseases can be improved by one optimal dose of DHEA or Astragalus. PMID- 19301630 TI - Batrachochytrium dendrobatidis, a novel pathogen approaching endemism in central California. AB - The recent emergence of amphibian chytridiomycosis has precipitated competing hypotheses regarding the endemic versus novel nature of the causative agent, Batrachochytrium dendrobatidis (Bd). We conducted a retrospective survey of the California Academy of Sciences' (San Francisco, California, USA) amphibian collection, testing for presence of Bd in 4 amphibian species collected from central California between 1897 and 2005. The earliest detection of Bd was found in 2 Rana catesbeiana in 1961, and the data support the hypothesis that Bd was a novel pathogen introduced into central California prior to 1961 that spread out geographically and taxonomically from at least one central location and is now endemic throughout most of central California. The taxonomic pattern of infection prevalence and the ecological constraints of the 4 species we tested suggest that, although Bd was initially detected in R. catesbeiana, the more efficient and most likely local vector for Bd in central California is actually Pseudacris regilla. PMID- 19301631 TI - Addition of antifungal skin bacteria to salamanders ameliorates the effects of chytridiomycosis. AB - Chytridiomycosis, caused by the skin fungus Batrachochytrium dendrobatidis (Bd), has caused population declines of many amphibians in remote protected habitats. Progress has been made in understanding the pathogen's life cycle, documenting its devastating effects on individual amphibians and on populations, and understanding how and why disease outbreaks occur. No research has directly addressed the critical question of how to prevent declines and extinctions caused by outbreaks of the disease. We have identified a number of bacterial species of amphibian skin that inhibit Bd in vitro. Here, we demonstrate that a species of anti-Bd skin bacteria can be successfully added to skins of salamanders Plethodon cinereus, and that addition of this bacterium reduced the severity of a disease symptom in experimentally infected individuals. This is the first demonstration that manipulating the natural skin microbiota of an amphibian species can alter the pathogen's negative effects on infected amphibians and appears to be the first demonstration that an epibiotic manipulation of any wildlife species can lessen the effects of an emerging infectious disease. It suggests that probiotic or bio-augmentation manipulations of cutaneous microbiota could have the potential to reduce susceptibility of amphibians to the disease in nature. This is the first approach suggested that could slow or halt epidemic outbreaks and allow successful reintroductions of amphibian species that have become locally or globally extinct in the wild. Our results also suggest a mechanism for the association of climate change and the likelihood of chytridiomycosis outbreaks via the effects of the former on antifungal bacterial communities. PMID- 19301632 TI - Prevalence of serum antibodies against Saprolegnia parasitica in wild and farmed brown trout Salmo trutta. AB - The prevalence of serum antibodies against Saprolegnia parasitica in wild and farmed brown trout Salmo trutta from the province of Le6n (NW Spain) was studied by enzyme-linked immunosorbent assay (ELISA). Blood samples from healthy and Saprolegnia-infected brown trout were collected over 2 yr with a seasonal periodicity (January, April, July and October) from a hatchery and river with frequent presence of saprolegniosis (River Porma) and from a river in which the disease was rarely observed (River Omana). The individual prevalence was 30.1%, but statistically significant differences were observed between the prevalence in trout from the hatchery (43.0%), from River Porma (31.8%) and from River Omana (6.4%) and also between the prevalence observed in October (42.9%) and the values obtained in January (24.8%), April (22.7%) and July (27.5%). There was no difference between the seroprevalence in females (34.8%) and males (38.2%), but a positive correlation between raised serum antibody levels and larger (older) fish was found. The low prevalence of antibodies observed in Saprolegnia-infected trout (18.0%) suggests possible immune suppression and the lack of an effective specific immune response in fish with saprolegniosis. PMID- 19301633 TI - Antibody response of rainbow trout with single or double infections involving viral haemorrhagic septicaemia virus and infectious haematopoietic necrosis virus. AB - Juvenile rainbow trout Oncorhynchus mykiss were experimentally infected by immersion with viral haemorrhagic septicaemia virus (VHSV), infectious haematopoietic necrosis virus (IHNV) or with both viruses. The presence of neutralizing antibodies in the sera of infected fish were analysed by 50% plaque neutralization tests (50%PNT). In Group 1 (infected with VHSV) and Group 2 (infected with IHNV) neutralizing antibodies were found in 41% and 21% of the serum samples, respectively. No cross-reacting antibodies were found in these 2 groups. In Group 3 (infected with both viruses) 30% of the samples showed neutralizing antibodies against VHSV, 21% against IHNV and 12% against both viruses. Fish in Group 3 developed a double specific antibody reaction whose kinetics and intensity (mean of log10 titres) were similar to the antibody response of the single infected groups. PMID- 19301634 TI - Copper sulfate toxicity to two isolates of Ichthyophthirius multifiliis relative to alkalinity. AB - Theronts from 2 different strains of Ichthyophthirius multifiliis (AR1 and AR5) were exposed to copper sulfate (CuSO4) in waters of different total alkalinities and observed for 4 h to determine relative toxicity and kinetics of parasite mortality. Consistent with the known solubility properties of the metal, Cu was significantly more toxic to cells maintained under low (48 mg l(-1)) compared with high (243 mg l(-1)) total alkaliniity conditions. This was reflected in both the median lethal concentration (LC50) values and rates of mortality for both parasite strains; strain differences were also observed. The AR1 strain was significantly more resistant to copper toxicity than the AR5 strain in both high and low alkalinity waters. In general, these strain differences were more evident under conditions of low stress (i.e. low CuSO4 concentration and high alkalinity), and suggest that genetic factors are overridden under high stress conditions. The present study establishes a role for alkalinity in the effectiveness of CuSO4 treatment of ichthyophthiriasis and reveals differences in the susceptibility of parasite populations that are clearly important for control programs. PMID- 19301635 TI - Myxozoan infections in fishes of the Tasik Kenyir Water Reservoir, Terengganu, Malaysia. AB - During a survey on fishes of the Tasik Kenyir Reservoir, Malaysia, 5 new Myxobolus spp. and 2 known Henneguya spp. were found. The specific locations for 2 Myxobolus spp. were the host's muscles, while 2 other Myxobolus spp. were found to develop in the host's kidney and gills, respectively. Of the species developing intracellularly in muscle cells, M. terengganuensis sp. nov. was described from Osteochilus hasselti and M. tasikkenyirensis sp. nov. from Osteochilus vittatus. M. csabai sp. nov. and M. osteochili sp. nov. were isolated from the kidney of Osteochilus hasselti, while M. dykovae sp. nov. was found in the gill lamellae of Barbonymus schwanenfeldii. Henneguya shaharini and Henneguya hemibagri plasmodia were found on the gills of Oxyeleotris marmoratus and Hemibagrus nemurus, respectively. Description of the new and known species was based on morphological characterization of spores, histological findings on locations of plasmodia and DNA sequence data. PMID- 19301636 TI - Myxozoan pathogens of Malaysian fishes cultured in ponds and net-cages. AB - We describe new myxosporean species from Malaysian fishes cultured in pond farms and net-cages. Myxobolus omari sp. nov. and M. leptobarbi sp. nov. were found in the muscles of Pangasianodon hypophthalmus and Leptobarbus hoevenii, respectively, while plasmodia and spores of Thelohanellus zahrahae sp. nov. and Henneguya daoudi sp. nov. were detected in the gills of Barbonymus gonionotus and Trichogaster trichopterus, respectively. Plasmodia and spores found in these fishes differed from the known myxosporean species in respect of their morphology, tissue tropism and 18S rDNA structure. No major pathological changes were found, but in the future these species might pose a potential threat to more intensified fish culture. PMID- 19301637 TI - Hematodinium sp. in Irish Cancer pagurus fisheries: infection intensity as a potential fisheries management tool. AB - Infection of Cancer pagurus by a parasitic dinoflagellate of the genus Hematodinium is described for the first time in Ireland. An industry-based monitoring programme was established to determine seasonality of infection intensity and prevalence in the country's 3 largest brown crab fisheries in the southwest, north and southeast. The parasite was present in all areas for the majority of sampling periods, with highest prevalences recorded in pre-recruit animals of both sexes. Microscopic examination of haemolymph revealed trophont, plasmodial and dinospore stages of the parasite. Overall prevalence in males (16%) was higher than in females (9%). Prevalence of Hematodinium sp. infection ranged from 0 to 51%, but a distinct seasonal trend was not apparent. Infection intensity was seasonal with significantly higher peaks occurring in late autumn/early winter months than in other quarters, corresponding to industry reports of moribund and dead pink-shelled crabs in commercial catches. We postulate that seawater temperature or a temperature-linked process is a key factor in triggering the final stages of infection, as significant autumn peaks were followed by a reduction in infection intensity as temperature decreased in the late winter/early spring months with no increase in intensity again until the following autumn. We propose that infection intensity, rather than prevalence, provides a more appropriate indication of the period when there is greatest potential for biological and economic impacts; the parameter's application as a fisheries management tool is discussed. PMID- 19301638 TI - Proliferative kidney disease in rainbow trout: time- and temperature-related renal pathology and parasite distribution. AB - Proliferative kidney disease is a parasitic infection of salmonid fishes caused by Tetracapsuloides bryosalmonae. The main target organ of the parasite in the fish is the kidney. To investigate the influence of water temperature on the disease in fish, rainbow trout Oncorhynchus mykiss infected with T bryosalmonae were kept at 12 degrees C and 18 degrees C. The number of parasites, the type and degree of lesions in the kidney and the mortality rate was evaluated from infection until full development of disease. While mortality stayed low at 12 degrees C, it reached 77% at 18 degrees C. At 12 degrees C, pathological lesions were dominated by a multifocal proliferative and granulomatous interstitial nephritis. This was accompanied by low numbers of T. bryosalmonae, mainly located in the interstitial lesions. With progression of the disease, small numbers of parasites appeared in the excretory tubuli, and parasite DNA was detected in the urine. Parasite degeneration in the interstitium was observed at late stages of the disease. At 18 degrees C, pathological lesions in kidneys were more severe and more widely distributed, and accompanied by significantly higher parasite numbers. Distribution of parasites in the renal compartments, onset of parasite degeneration and time course of appearance of parasite DNA in urine were not clearly different from the 12 degrees C group. These findings indicate that higher mortality at 18 degrees C compared to 12 degrees C is associated with an enhanced severity of renal pathology and increased parasite numbers. PMID- 19301639 TI - Etiology and pathology of epidermal papillomas in allogynogenetic crucian carp Carassius auratus gibelio (female) x Cyprinus carpio var. singuonensis (male). AB - Allogynogenetic crucian carp Carassius auratus gibelio (female) x Cyprinus carpio var. singuonensis (male) is one of the main freshwater aquaculture species in China. In recent years, epidermal papillomas have been observed on the scales, fins and opercula of adult fish in many fish farms in the Chongming county of Shanghai, China. The disease appears in the late autumn of the first year and becomes more severe in winter. It gradually regresses in the late spring or summer of the second year, as water temperature increases. Our study revealed that the disease pathogen was likely to be a herpes-like virus, as indicated by enveloped viral particles in the cytoplasm, empty capsids in the nucleus and a virus-like morphology of the pathogen. The size of the enveloped herpes-like virus was 118.18 +/- 10.53 (SD) nm (n = 22) and its nucleocapsid was 78.64 +/- 7.74 nm (n = 22) in diameter. Histopathological examination of tumours revealed that both epithelial and stromal cells proliferated to form papillomas. The nuclei of epithelial tumour cells were irregular in shape and in size. Most of the mucous cells were located in clusters near the middle of each papilloma. Cytoplasmic organelles were sparse in tumour cells. Numerous granulocytes and lymphocytes infiltrated into the tumour tissue. There were no inclusion bodies in the cytoplasm and nuclei of tumour cells. The lesions only occurred in adult allogynogenetic crucian carp, even though they were cultured together with other fish species. There was marked variation in incidence: in some ponds, only a few fish were affected, while the incidence was up to 90% in other ponds. PMID- 19301640 TI - Methods for selecting effective siRNA sequences by using statistical and clustering techniques. AB - Short interfering RNAs (siRNAs) have been widely used for studying gene functions in mammalian cells but vary markedly in their gene-silencing efficacy. Although many design rules/guidelines for effective siRNAs based on various criteria have been reported recently, there are only a few consistencies among them. This makes it difficult to select effective siRNA sequences targeting mammalian genes. This chapter first reviews the reported siRNA design guidelines and clarifies the problems concerning the current guidelines. It then describes the recently reported new scoring methods for selecting effective siRNA sequences by using statistics and clustering techniques such as the self-organizing map (SOM) and the radial basis function (RBF) network. In the proposed three methods, individual scores are defined as a gene degradation measure based on position specific statistical significances. The effectiveness of the methods was confirmed by evaluating effective and ineffective siRNAs for recently reported genes and comparison with other reported scoring methods. The sizes (values) of these scores are closely correlated with the degree of gene degradation, and the scores can easily be used for selecting high-potential siRNA candidates. The evaluation results indicate that the proposed new methods are useful for selecting siRNA sequences targeting mammalian mRNA sequences. PMID- 19301641 TI - Deciphering the code of innate immunity recognition of siRNAs. AB - Small interfering RNAs (siRNAs) have been widely used for knocking down gene expression in a variety of organisms. Although experiments in cancer cell lines indicate that siRNAs are usually not detected by innate immunity, lipid-mediated delivery of siRNAs into blood cells is often accompanied by the activation of immunity. Recent studies indicated that certain siRNA sequences engage Toll-like receptor TLR7/8 signalling resulting in the activation of a large number of host defense genes including interferons (IFNs), proinflammatory cytokines, Mx proteins, chemokines, chemokine receptors, costimulatory molecules, RNA helicases, galectins, and ubiqitin ligases. In addition to immune activation, most siRNA sequences, if not all, can silence multiple genes in addition to the intended target gene, a phenomenon known as "off-target effects." Hence, one of the major challenges for therapeutic applications of siRNAs is to decipher the mechanisms involved in siRNA recognition by the immune system and to identify strategies that can evade immune activation. In this respect, the replacement of only uridines with their 2'-modified counterparts such as 2'-O-methyl uridines abrogates immune recognition of siRNAs. Interestingly, 2'-O-methyl-modified RNAs not only evade TLR7/8-sensing pathways, but also reduce siRNA off-target effects and antagonize with a variety of immunostimulatory RNAs to activate TLR7/8 signalling. RNA oligonucleotides and duplex siRNAs with 2'-deoxy uridines or thymidines exhibited no significant immunostimulatory effects and binding potency to TLRs. Therefore, I recommend the use of these modifications in order to evade immune sensing of siRNA and off-target effects. This chapter addresses the current state of knowledge regarding the molecular and cellular mechanisms of RNA recognition by the immune system and proposes a range of strategies allowing the design of siRNAs with minimal or maximal immunostimulatory potency for therapeutic applications. PMID- 19301642 TI - Targeted delivery of antisense oligonucleotides and siRNAs into mammalian cells. AB - RNA interference (RNAi) is a natural mechanism for gene silencing that can be harnessed for the development of RNA-based drugs. Although synthetic small interfering RNA (siRNAs) can be delivered in vitro to virtually all cell types using lipid-based transfection agents or electroporation, efficient strategies for achieving either systemic or targeted delivery remains one of the major in vivo challenges. Among the targeting strategies, receptor-targeted delivery provides an innovative strategy to selectively direct therapeutics to cancer cells. Receptor-binding peptides can be incorporated into gene-delivery vesicles or directly conjugated to siRNAs in the hope of promoting their localization in target cells expressing the cognate receptors. This chapter discusses the current status of siRNA-targeting strategies using either peptides identified through iterative screening of random peptide phage libraries or naturally occurring peptides. Also, transcriptional targeting strategies and detailed protocols for the selection of cancer cell-binding peptide from random peptide libraries are described. PMID- 19301643 TI - Local and systemic delivery of siRNAs for oligonucleotide therapy. AB - RNA interference (RNAi) is a relatively new found phenomenon of posttranscriptional gene silencing to regulate the expression of multiple genes involved in a wide range of biological processes. The gene-silencing technology via RNAi has also been developed into a commonly anti-gene method. Furthermore, in vivo data indicate that small interfering RNAs (siRNAs) may be used to treat human diseases. However, the most challenging issue to a successful in vivo application is the development of a delivery system that can transport siRNA molecules into the tissues and/or the cells of interest. Also, the evaluation of siRNA potency in vivo is central for the selection of therapeutic siRNAs. In this chapter, the effects of atelocollagen-delivered siRNAs in live animals were monitored using bioluminescence imaging. PMID- 19301644 TI - Imaging of siRNA delivery and silencing. AB - The fast developing field of RNA interference (RNAi) requires monitoring of small interfering RNA (siRNA) delivery to targeted organs and evaluating the efficiency of target gene silencing. The molecular imaging approach fits perfectly to fulfill these needs and provides information in a fast, reproducible, and noninvasive manner. This review serves as a first attempt to summarize existing information on various imaging modalities and their application for siRNA imaging. It is noteworthy that new publications in this field appear almost on a weekly basis and the authors have made a sincere attempt to reflect the development of this area in their review. PMID- 19301645 TI - Recent advances in magnetofection and its potential to deliver siRNAs in vitro. AB - This chapter describes how to design and conduct experiments to deliver siRNA to adherent mammalian cells in vitro by magnetic force-assisted transfection using self-assembled complexes of small interfering RNA (siRNA) and cationic lipids or polymers that are associated with magnetic nanoparticles. These magnetic complexes are targeted to the cell surface by the application of a magnetic gradient field. In this chapter, first we describe the synthesis of magnetic nanoparticles for magnetofection and the association of siRNA with the magnetic components of the transfection complex. Second, a simple protocol is described in order to evaluate magnetic responsiveness of the magnetic siRNA transfection complexes and estimate the complex loading with magnetic nanoparticles. Third, protocols are provided for the preparation of magnetic lipoplexes and polyplexes of siRNA, magnetofection, downregulation of gene expression, and the determination of cell viability. The addition of INF-7 peptide, a fusogenic peptide, to the magnetic transfection triplexes improved gene silencing in HeLa cells. The described protocols are also valuable for screening vector compositions and novel magnetic nanoparticle preparations to optimize siRNA transfection by magnetofection in every cell type. PMID- 19301646 TI - In vitro and in vivo gene silencing by TransKingdom RNAi (tkRNAi). AB - RNA interference (RNAi) is a potent and specific mechanism for eliminating the mRNA of specific genes. This gene silencing mechanism occurs naturally and is highly conserved from plants to human cells, holding promise for functional genomics and for revolutionizing medicine due to its unlimited potential to treat genetic, epigenetic, and infectious disease. However, efforts to unleash the enormous potential of RNAi have met with significant challenges. Delivery is problematic because short interfering RNAs (siRNA) are negatively charged polymers that inefficiently enter cells and undergo rapid enzymatic degradation in vivo. In addition, the synthesis of siRNAs is expensive for long-term research and therapeutic applications. Recently, we have shown that nonpathogenic bacteria can be engineered to activate RNAi in mammalian cells (TransKingdom RNA interference; tkRNAi). This new approach offers several advantages and has significant implications. First, this method allows the establishment of a long term stable gene silencing system in the laboratory against genes of interests in vitro and in vivo, and enables high-throughput functional genomics screening in mammalian systems. RNAi libraries can be constructed, stored, reproduced, amplified, and used with the help of E. coli as currently done with gene cloning. Second, this technology provides a clinically compatible way to achieve RNAi for therapeutic applications due to the proven clinical safety ofnonpathogenic bacteria as a gene carrier, tkRNAi also eliminates the siRNA manufacture issue, and may circumvent or mitigate host interferon-like responses since siRNA is produced intracellularly. PMID- 19301647 TI - Bacterial delivery of siRNAs: a new approach to solid tumor therapy. AB - RNAi is a powerful research tool for specific gene silencing and may also lead to promising novel therapeutic strategies. However, the development of RNAi-based therapies has been slow due to the lack of targeted delivery methods. The biggest challenge in the use of siRNA-based therapies is the delivery to target cells. There are many additional obstacles to in vivo delivery of siRNAs, such as degradation by endogenous enzymes and interaction with blood components leading to nonspecific uptake into cells, which govern biodistribution and availability of siRNA in the body. Naked unmodified synthetic siRNA including plasmid-carried shRNA-expression constructs cannot penetrate cellular membranes, and therefore, systemic application is unlikely to be successful. The success of gene therapy by siRNAs relies on the development of safe, economical, and efficacious in vivo delivery systems into the target cells. Attenuated Salmonella have been employed recently as vectors to deliver silencing hairpin RNA (shRNA) expression plasmids into mammalian cells. This approach has achieved gene silencing in vitro and in vivo. The facultative anaerobic, invasive Salmonella have a natural tropism for solid tumors including metastatic tumors. Genetically modified, attenuated Salmonella have been used recently both as potential antitumor agents by themselves, and to deliver specific tumoricidal therapies. This chapter describes the use of attenuated bacteria as tumor-targeting delivery systems for cancer therapy. PMID- 19301648 TI - The therapeutic potential of LNA-modified siRNAs: reduction of off-target effects by chemical modification of the siRNA sequence. AB - Post-transcriptional gene silencing mediated by double-stranded RNA represents an evolutionarily conserved cellular mechanism. Small dsRNAs, such as microRNAs (miRNAs), are part of the main regulatory mechanisms of gene expression in cells. The possibilities of harnessing this intrinsic natural mechanism of gene silencing for therapeutic applications was opened up by the discovery by Tom Tuschl's team a few years ago that chemically synthesized small 21-mers of double stranded RNA (small interfering RNA, siRNA) could inhibit gene expression without induction of cellular antiviral-like responses, siRNAs are especially of interest for cancer therapeutics because they allow specific inhibition of mutated oncogenes and other genes that aid and abet the growth of cancer cells. However, recent insights make it clear that siRNA faces some major hurdles before it can be used as a drug. Some of these problems are similar to those associated with classic antisense approaches, such as lack of delivery to specific tissues (other than the liver) or tumors, while other problems are more specific for siRNA, such as stability of the RNA molecules in circulation, off-target effects, interference with the endogenous miRNA machinery, and immune responses toward dsRNA. Chemical modifications of siRNA may help prevent these unwanted side effects. Initial studies show that minimal modifications with locked nucleic acids (LNA) help to reduce most of the unwanted side effects. In this chapter we will explore the limitations and possibilities of LNA-modified siRNA that may be used in future therapeutic applications. PMID- 19301649 TI - pSM155 and pSM30 vectors for miRNA and shRNA expression. AB - MicroRNAs (miRNAs) have key roles in diverse regulatory pathways, including control of developmental timing, cell differentiation, apoptosis, cell proliferation, and organ development. miRNAs regulate gene function through a process termed RNA interference (RNAi), which is a highly conserved intracellular mechanism that regulates posttranscriptional gene silencing. RNAi is triggered by double-stranded small interfering RNAs (siRNAs), which can be processed from small hairpin RNAs (shRNAs) generated from an expression vector. In some recently described vectors, the siRNAs are expressed from synthetic stem-loop precursors of microRNAs (miRNAs) driven by polymerase II promoters. We have reported new RNAi vectors, designated pSM155 and pSM30, that take into consideration miRNA processing and RNA splicing by placing the miRNA-based artificial miRNA expression cassettes inside of synthetic introns. Like the original miRNA vectors, these pSM155 and pSM30 constructs can efficiently downregulate the expression of their target genes. Moreover, the expression of a coexpressed fluorescent marker, EGFP, is substantially improved by this new design. The new vectors can also be used to express natural miRNAs and label cells expressing these miRNAs. These RNAi vectors thus provide new tools for gene suppression and miRNA expression. We describe in this chapter the protocols for selecting and cloning artificial and natural miRNAs (or shRNAs), evaluating their efficiency in downregulating gene expression, and also discuss the potential applications of these vectors. PMID- 19301650 TI - Targeting oncogenes with siRNAs. AB - Current cancer chemotherapies heavily rely on the unspecific inhibition of proliferating cells. This lack of tumour cell specificity results in severe toxic side effects and may only hardly affect quiescent cancer stem cells consequently leading to relapse. Since oncogenes are exclusively expressed in malignant and pre-malignant cells, they may provide unique, cancer cell specific targets for therapeutic strategies. However, their role in maintaining the malignant phenotype is frequently unknown. Furthermore, oncogenic transcription factors are generally considered to be "undruggable" with conventional small molecule approaches. Oncogene-specific RNA interference offers here new and exciting options to analyse oncogene functions directly in the malignant environment. Moreover, such approaches may permit the targeting of oncogenic transcription factors, thereby considerably extending the number of cancer-specific target structures. In this chapter, several rationales and practical aspects of oncogene targeting with siRNAs are discussed. Special emphasis is given to the application of RNA interference to haematopoietic cells, which are generally hard to transfect. In particular, solving the problem of systemic siRNA/shRNA delivery will greatly advance the inclusion of RNA interference strategies into more efficient and specific therapeutic strategies. PMID- 19301651 TI - Targeting stromal-cancer cell interactions with siRNAs. AB - Tumors are composed of both malignant and normal cells, including fibroblasts, endothelial cells, mesenchymal stem cells, and inflammatory immune cells such as macrophages. These various stromal components interact with cancer cells to promote growth and metastasis. For example, macrophages, attracted by colony stimulating factor-1 (CSF-1) produced by tumor cells, in turn produce various growth factors such as vascular endothelial growth factor, which supports the growth of tumor cells and their interaction with blood vessels leading to enhanced tumor cell spreading. The activation of autocrine and paracrine oncogenic signaling pathways by stroma-derived growth factors and cytokines has been implicated in promoting tumor cell proliferation and metastasis. Furthermore, matrix metalloproteinases (MMPs) derived from both tumor cells and the stromal compartment are regarded as major players assisting tumor cells during metastasis. Collectively, these recent findings indicate that targeting tumor-stroma interactions is a promising strategy in the search for novel treatment modalities in human cancer. This chapter summarizes our current understanding of the tumor microenvironment and highlights some potential targets for therapeutic intervention with small interfering RNAs. PMID- 19301652 TI - Therapeutic potential of siRNA-mediated targeting of urokinase plasminogen activator, its receptor, and matrix metalloproteinases. AB - Targeting proteases and their activators would retard the invasive ability of cancer cells, and has been shown to induce apoptosis in certain instances. Various methods have been developed to specifically target protease molecules in an attempt to retard invasion and migration. Of these methods, RNA interference (RNAi) holds great therapeutic potential. RNAi technology is now being used to target specific molecules for use as potential anti-cancer agents. RNAi-mediated silencing is almost catalytic when compared to anti-sense silencing. Of these targets, the uPAR-uPA system and MMPs holds great promise. Targeting uPA/uPAR may provide additive or synergistic treatment benefits if used in combination with conventional therapeutics such as chemotherapy or radiation. Studies point to the fact that specifically targeting MMP-9 or MMP-2 singly or in combination with other proteases could have specific therapeutic implications in the treatment of cancer. In this chapter we discuss the therapeutic potential of siRNA-mediated targeting of the uPAR-uPA system and MMPs as therapeutic agents for the treatment of cancer. PMID- 19301653 TI - Silencing of HIF-1alpha by RNA interference in human glioma cells in vitro and in vivo. AB - Higher-grade gliomas are distinguished by increased vascular endothelial cell proliferation and peritumoral edema. These are thought to be instigated by vascular endothelial growth factor, which in turn is regulated by cellular oxygen tension. Hypoxia inducible factor-1alpha (HIF-1alpha) is a main responder to intracellular hypoxia and is overexpressed in many human cancers, including gliomas. Here we present methods for investigating the role of HIF-1alpha in glioma growth in vivo and in vitro using RNA interference in U251, U87, and U373 glioma cells. PMID- 19301654 TI - RNA interference-mediated validation of genes involved in telomere maintenance and evasion of apoptosis as cancer therapeutic targets. AB - The discovery of new cancer-related therapeutic targets is mainly based on the identification of genes involved in pathways selectively exploited in cancer cells, including those leading to unlimited replicative potential, evasion of apoptosis, angiogenesis, tissue invasion and metastatic spread. Potentially, a gene--or a gene product--is recognized as a cancer target whether its modulation in experimental models can specifically modify or revert the cancer phenotype. As soon as RNA interference (RNAi)--a natural gene silencing mechanism--was demonstrated in mammalian cells, it rapidly became an essential means for gene knockdown in preclinical models, making it possible to define the role of several human genes and to identify those specifically involved in the onset and progression of cancer. Owing to its powerful gene-silencing properties, RNAi has been proposed as a useful tool to validate new therapeutic targets and to develop innovative anticancer therapies. This chapter summarizes the findings from recent studies relying on the use of RNAi-based approaches to functionally validate therapeutic targets related to two tumor hallmarks: the unlimited replicative potential (i.e., activation of telomere maintenance mechanisms) and evasion of apoptosis (i.e., up-regulation of anti-apoptotic factors). PMID- 19301655 TI - Treating respiratory viral diseases with chemically modified, second generation intranasal siRNAs. AB - Chemically synthesized short interfering RNA (siRNA) of pre-determined sequence has ushered a new era in the application of RNA interference (RNAi) against viral genes. We have paid particular attention to respiratory viruses that wreak heavy morbidity and mortality worldwide. The clinically significant ones include respiratory syncytial virus (RSV), parainfluenza virus (PIV) and influenza virus. As the infection by these viruses is clinically restricted to the respiratory tissues, mainly the lungs, the logical route for the application of the siRNA was also the same, i.e., via the nasal route. Following the initial success of intranasal siRNA against RSV, second-generation siRNAs were made against the viral polymerase large subunit (L) that were chemically modified and screened for improved stability, activity and pharmacokinetics. 2'-O-methyl (2'-O-Me) and 2' deoxy-2'-fluoro (2'-F) substitutions in the ribose ring were incorporated in different positions of the sense and antisense strands and the resultant siRNAs were tested with various transfection reagents intranasally against RSV. Based on these results, we propose the following consensus for designing intranasal antiviral siRNAs: (i) modified 19-27 nt long double-stranded siRNAs are functional in the lung, (ii) excessive 2'-OMe and 2'-F modifications in either or both strands of these siRNAs reduce efficacy, and (iii) limited modifications in the sense strand are beneficial, although their precise efficacy may be position dependent. PMID- 19301656 TI - Progress in the therapeutic applications of siRNAs against HIV-1. AB - Therapeutic options against the human immunodeficiency virus type 1 (HIV-1) continue to expand with the development of new drugs and new therapeutic strategies. Nevertheless, management of HIV-1 infected individuals has become increasingly complex. The emergence of drug-resistant variants, the growing recognition of the long-term toxicity of antiretroviral therapies and the persistence of viral reservoirs justify the continued efforts to develop new anti HIV-1 strategies. Recent advances regarding the utility of RNA-mediated interference (RNAi) to specifically inhibit HIV-1 replication have opened new possibilities for the development of gene-based therapies against HIV-1 infection. Here, the recent advances in siRNA-based therapies are reviewed. PMID- 19301657 TI - Protein components of the microRNA pathway and human diseases. AB - MicroRNAs (miRNAs) are key regulators of messenger RNA (mRNA) translation known to be involved in a wide variety of cellular processes. In fact, their individual importance is reflected in the diseases that may arise upon the loss, mutation or dysfunction of specific miRNAs. It has been appreciated only recently that diseases may also develop when the protein components of the miRNA machinery itself are affected. The core enzymes of the major protein complexes involved in miRNA biogenesis and function, such as the ribonucleases III (RNases III) Drosha and Dicer as well as Argonaute 2 (Ago2), appear to be essential. However, the accessory proteins of the miRNA pathway, such as the DiGeorge syndrome critical region gene 8 (DGCR8) protein, Exportin-5 (Exp-5), TAR RNA binding protein (TRBP) and fragile X mental retardation protein (FMRP), are each related, in various ways, to specific genetic diseases. PMID- 19301658 TI - Intron-mediated RNA interference and microRNA biogenesis. AB - Nearly 97% of the human genome is non-coding DNA, and introns occupy most of it around the gene-coding regions. Numerous intronic sequences have been recently found to encode microRNAs, which are responsible for RNA-mediated gene silencing through RNA interference (RNAi)-like pathways. microRNAs (miRNAs), small single stranded regulatory RNAs capable of interfering with intracellular messenger RNAs (mRNAs) that contain either complete or partial complementarity, are useful for the design of new therapies against cancer polymorphism and viral mutation. This flexible characteristic is different from double-stranded siRNAs (small interfering RNAs) because a much more rigid complementarity is required for siRNA induced RNAi gene silencing. miRNAs were firstly discovered in Caenorhabditis elegans as native RNA fragments that modulate a wide range of genetic regulatory pathways during embryonic development. Currently, varieties of miRNAs are widely reported in plants, animals and even microbes. Intronic microRNA is a new class of miRNAs derived from the processing of gene introns. The intronic miRNAs differ uniquely from previously described intergenic miRNAs in the requirement of type II RNA polymerases (Pol-II) and spliceosomal components for their biogenesis. Several kinds of intronic miRNAs have been identified in C. elegans, mouse and human cells; however, neither function nor application has been reported. Here, we show for the first time that intron-derived miRNAs are able to induce RNA interference in not only human and mouse cells but also zebrafishes, chicken embryos and adult mice, demonstrating the evolutionary preservation of the intron mediated gene silencing through miRNA functionality in cell and in vivo. These findings suggest an intracellular miRNA-mediated gene regulatory system, fine tuning the degradation of protein-coding messenger RNAs. PMID- 19301659 TI - Emergence of a complex relationship between HIV-1 and the microRNA pathway. AB - Recent experimental evidences support the existence of an increasingly complex and multifaceted interaction between viruses and the microRNA-guided RNA silencing machinery of human cells. The discovery of small interfering RNAs (siRNAs), which are designed to mediate cleavage of specific messenger RNAs (mRNAs), prompted virologists to establish therapeutic strategies based on siRNAs with the aim to suppress replication of several viruses, including human immunodeficiency virus type 1 (HIV-1). It has been appreciated only recently that viral RNAs can also be processed endogenously by the microRNA-generating enzyme Dicer or recognized by cellular miRNAs, in processes that could be viewed as an adapted antiviral defense mechanism. Known to repress mRNA translation through recognition of specific binding sites usually located in their 3' untranslated region, miRNAs of host or viral origin may exert regulatory effects towards host and/or viral genes and influence viral replication and/or the host response to viral infection. This article summarizes our current state of knowledge on the relationship between HIV-1 and miRNA-guided RNA silencing, and discusses the different aspects of their interaction. PMID- 19301660 TI - Synthetic microRNA targeting glioma-associated antigen-1 protein. AB - The transcription factor glioma-associated antigen-1 (Gli-1) mediates activation of the sonic hedgehog (Shh) pathway, a process that precedes the transformation of tissue stem cells into cancerous stem cells and that is involved in early and late epithelial tumorigenesis. Hypothesizing that targeting the 3'-untranslated region (3'-UTR) of Gli-1 mRNA would effectively inhibit epithelial tumor cell proliferation, we evaluated several complementary miRNA molecules for their ability to do so. The synthetic miRNAs and corresponding duplex/small temporal RNAs were introduced as 3-nucleotide (nt) loops into GU-rich portions of the 3'UTR Gli-1 sequence. One particular miRNA (miRNA Gli-1-3548) and its corresponding duplex (Duplex 3548) significantly inhibited proliferation of Gli 1+ ovarian (SK-OV-3) and pancreatic (MiaPaCa-2) tumor cells by delaying cell division and activating late apoptosis in MiaPaCa-2 cells. Here, we describe the design of effective miRNA sequences and their applications as anti-gene agents. PMID- 19301661 TI - Therapeutic targeting of gene expression by siRNAs directed against BCR-ABL transcripts in a patient with imatinib-resistant chronic myeloid leukemia. AB - Within the recent years, RNA interference (RNAi) has become an almost-standard method for in vitro knockdown of any target gene of interest. Now, one major focus is to further explore its potential in vivo, including the development of novel therapeutic strategies. From the mechanism, it becomes clear that small interfering RNAs (siRNAs) play a pivotal role in triggering RNAi. Thus, the efficient delivery of target gene-specific siRNAs is one major challenge in the establishment of therapeutic RNAi. Here we show that in vivo application of targeted nonvirally delivered synthetic bcr-abl siRNA in a female patient with recurrent Philadelphia chromosome positive chronic myeloid leukemia (CML) resistant to imatinib (Y253F mutation) and chemotherapy after allogeneic hematopoietic stem cell transplantation can silence the expression of bcr-abl gene. We found a remarkable inhibition of the overexpressed bcr-abl oncogene resulting in increased apoptosis of CML cells. In vivo siRNA application was well tolerated without any clinically adverse events. Our findings imply that the clinical application of synthetic siRNA is feasible, safe and has real potential for genetic-based therapies using synthetic nonviral carriers. PMID- 19301662 TI - Angiogenic signalling pathways. AB - Hypoxia is widely recognised as a key driving force for tumor angiogenesis by its induction of vascular endothelial growth factor (VEGF) and other direct-acting angiogenic factors. We describe the effect of hypoxia on gene expression and downstream angiogenic signalling; however, the angiogenic process is complex, and many other signalling pathways beyond VEGF are implicated in the formation of new vessels. These include extra-cellular signalling pathways such as the notch/delta, ephrin/Eph receptor, roundabout/slit, and netrin/UNC (uncoordinated) receptor families as well as intracellular proteins such as hedgehog and sprouty. The remarkable diversity in angiogenic signalling pathways provides many opportunities for therapeutic intervention, and anti-angiogenesis is currently a major area of oncology research. PMID- 19301663 TI - Imaging angiogenesis. AB - There is a need for direct imaging of effects on tumor vasculature in assessment of response to antiangiogenic drugs and vascular disrupting agents. Imaging tumor vasculature depends on differences in permeability of vasculature of tumor and normal tissue, which cause changes in penetration of contrast agents. Angiogenesis imaging may be defined in terms of measurement of tumor perfusion and direct imaging of the molecules involved in angiogenesis. In addition, assessment of tumor hypoxia will give an indication of tumor vasculature. The range of imaging techniques available for these processes includes positron emission tomography (PET), dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), perfusion computed tomography (CT), and ultrasound (US). PMID- 19301664 TI - Assessing tumor angiogenesis in histological samples. AB - Tumor neovascularization acquires vessels through a number of processes, including angiogenesis, vasculogenesis, vascular remodelling, intussusception, and possibly vascular mimicry in certain tumors. The end result of the tumor vasculature has been quantified by counting the number of immunohistochemically identified microvessels in areas of maximal vascularity so-called hot spots. Other techniques have been developed, such as Chalkley counting and the use of image analysis systems that are robust and reproducible as well as more objective. Many of the molecular pathways that govern tumor neovascularization have been identified, and many reagents are now available to study these tissue sections. These include angiogenic growth factors and their receptors, cell adhesion molecules, proteases, and markers of activated, proliferating, cytokine stimulated, or angiogenic vessels, such as CD105. It is also possible to differentiate quiescent from active vessels. Other reagents that can identify proteins involved in microenvironmental influences such as hypoxia have also been generated. Although the histological assessment of tumor vascularity is used mostly in the research context, it may also have clinical applications if appropriate methodology and trained observers perform the studies. PMID- 19301665 TI - Immunohistochemical methods for measuring tissue lymphangiogenesis. AB - The field of lymphatic research has benefited enormously from the recent discovery of "marker" proteins that permit not only the identification and quantitation of lymphatic vessels in tissue sections for tumor pathology but also the isolation of primary lymphatic endothelial cells for basic research. This chapter focuses on the use of these markers for the immunohistochemical analysis of lymphangiogenesis in both frozen and paraffin-embedded tissue sections and discusses current protocols and their associated problems. PMID- 19301666 TI - Vascular endothelial cells from human micro- and macrovessels: isolation, characterisation and culture. AB - The endothelium residing in different vascular beds displays high-degree phenotypic heterogeneity at morphological, functional, biochemical, and molecular levels. Endothelial cells (ECs) can be easily harvested from large vessels by mechanical removal or collagenase digestion. In particular, the umbilical vein has been used due to its wide availability, and study of the ECs derived from it have undoubtedly greatly advanced our knowledge of vascular biology. However, the majority of the body's endothelium (> 95%) forms the microvasculature, and it is these cells providing the interface between the blood and tissues that play a critical role in the development of new blood vessels. This has led to the establishment of techniques for the isolation of microvascular ECs from different tissues to provide more physiologically relevant in vitro models of angiogenesis and EC function. The main focus of this chapter is the use of superparamagnetic beads (Dynabeads) coupled to anti- platelet endothelial cell adhesion molecule-1 (PECAM-1) antibodies (PECA beads) to isolate microvessel ECs from human adipose and methods for the characterisation and maintenance of ECs in culture. Adipose tissue is an ideal source of microvessel ECs as it is composed mainly of adipocytes with a rich microvasculature and is easy to disaggregate. Adipose obtained at reduction mammoplasty or adominoplasty is first dissected free of connective tissue and subjected to collagenase type II digestion. The adipocytes are removed by several rounds of centrifugation and separated from the microvessel-rich pellet, which is then further disaggregated with trypsin/EDTA (ethylenediaminetetraacetic acid) solution. Following filtration to remove fragments of connective tissue, the pellet is incubated with PECA beads, and microvessel fragments/ECs are washed and harvested using a magnet. In addition, the adaptation of this basic technique to isolate microvessel ECs from human lung and stomach is also described along with methods for the preparation of large vessel endothelial cells. PMID- 19301667 TI - Lymphatic endothelial cells: establishment of primaries and characterization of established lines. AB - This chapter describes detailed methods for the isolation of primary human lymphatic endothelial cells from neonatal foreskin. We also provide protocols and information for their characterization and propagation. Isolation of primary human lymphatic endothelial cells requires a two-step process: mechanical and enzymatic digestion of human foreskins and cell sorting by fluorescence-activated cell sorting of CD31+/podoplanin+/CD45- cells. Characterization of these cells requires an assessment of the expression of several markers specific for lymphatic endothelium. This is determined by fluorescence-activated cell sorting, immunocytochemistry, and polymerase chain reaction. All procedures are based on simple laboratory techniques and, with the exception of a cell sorter and the skills to use it, do not require specialized equipment. PMID- 19301668 TI - Isolation, culture and characterisation of vascular smooth muscle cells. AB - Smooth muscle cells (SMCs) are key players in the pathogenesis of atherosclerosis and restenosis; however, they are also important in formation and development of de novo blood vessels during vasculogenesis and angiogenesis. Vascular SMCs can be formed by proliferation of existing SMCs, maturation of pericytes, or putative smooth muscle progenitor cells, thereby contributing to development of atherosclerotic plaques and angiogenic processes. Modulation of SMC phenotype is now recognised as a key event in the development of vascular diseases. This chapter describes the isolation and culture of vascular SMCs and pericytes from human and animal blood vessels for in vitro studies. PMID- 19301669 TI - Circulating endothelial progenitor cells. AB - Recent studies have found that bone marrow-derived cells give rise to endothelial cells during states of tissue repair and disease. We have found that one key integrin, integrin-alpha4beta1, promotes the homing of circulating endothelial progenitor cells (EPCs) to sites of ongoing tissue repair. This integrin facilitates the adhesion of EPCs to the vascular endothelium in inflamed tissue or within tumors. We demonstrate how to identify, isolate, purify, and characterize EPCs. We also demonstrate in vivo analysis of the roles of bone marrow-derived cells in tumor growth and angiogenesis by demonstrating adoptive transfer, bone marrow transplantation, tumor models, and immunohistochemistry for markers of blood and endothelial vessels. Finally, we show how to characterize cell adhesion mechanisms regulating bone marrow-derived progenitor cell trafficking. PMID- 19301670 TI - In vitro assays for endothelial cell functions related to angiogenesis: proliferation, motility, tubular differentiation, and proteolysis. AB - This chapter covers the breakdown of the process of angiogenesis into simple assays to measure discrete endothelial cell functions. The techniques described are suitable for studying stimulators or inhibitors of angiogenesis and determining which aspect of the process is modulated. The procedures outlined are robust and straightforward but cannot cover the complexity of the angiogenic process as a whole, incorporating as it does myriad positive and negative signals, three-dimensional interactions with host tissues and many accessory cells, including fibroblasts, macrophages, pericytes, and platelets. The extent to which in vitro assays predict responses in vivo (e.g., wound healing, tumor angiogenesis, or surrogate techniques such as Matrigel plugs, sponge implants, corneal assays, etc.) remains to be determined. PMID- 19301671 TI - Tube formation: an in vitro matrigel angiogenesis assay. AB - Neovascularization plays a role in several pathological conditions, including tumor growth, arthritis, and choroidal neovascularization. Investigators from different fields can choose from several available angiogenesis assays according to their specific needs. This chapter describes an easy-to-perform assay that is based on the differentiation of endothelial cells and the formation of tube-like structures on an extracellular matrix, Matrigel. The assay can be used to screen compounds for angiogenic activity or to determine if it has an effect on angiogenesis, depending on the conditions chosen. It is a quick assay, easy to set up, and highly reproducible. It can be used to test one or two samples, or it can quickly be scaled up to screen hundreds of compounds. The flexibility that this assay provides makes it a good first choice to test if a compound or a series of compounds may play a role in angiogenesis. PMID- 19301672 TI - Three-dimensional in vitro anglogenesis in the rat aortic ring model. AB - Angiogenesis is a complex sequential process involving endothelial activation, basement membrane degradation, endothelial sprouting from the parent vessel, invasion of the extracellular matrix, endothelial proliferation, vessel elongation, branching, anastomosis, increases in vessel diameter, basement membrane formation, pericyte acquisition, and remodelling. Most in vitro angiogenesis assays are two-dimensional and measure only one facet of this process, generally endothelial proliferation, migration, or tube formation. The two-dimensional nature of the assays also ignores the differences in endothelial phenotype seen in three-dimensional models and in vivo. The in vitro serum-free three-dimensional rat aortic model closely approximates the complexities of angiogenesis in vivo, from endothelial activation to pericyte acquisition and remodelling, and most of these can be quantified by image analysis, immunohistochemistry, and biochemical analysis. It is easily manipulated using molecular biological intervention or exogenous inhibitors and activators in a relatively controlled system. PMID- 19301673 TI - Static and dynamic assays of cell adhesion relevant to the vasculature. AB - Methods are described for analysing adhesion of isolated cells (such as leucocytes, tumor cells, or precursor cells) to purified adhesion receptors or cultured endothelial cells. "Static" assays (in which cells are allowed to settle on the adhesive substrates) and flow-based assays (in which cells are perfused over the substrates) are compared. Direct observations of the time course of adhesion and migration can be made when purified proteins or endothelial cells are cultured in plates, after cells are allowed to settle onto them for a desired period. In the flow-based assay, cells are perfused through coated glass capillaries or flow channels incorporating coated plates. Again, direct video microscopic observations are made. In this assay, various stages of capture, immobilisation and migration can be followed. In general, the static systems have higher throughput and greatest ease of use, but yield less-detailed information, while the flow-based assay is most difficult to set up but is most physiologically relevant if one is interested in the dynamics of adhesion in the vasculature. PMID- 19301674 TI - Genetic manipulation of corneal endothelial cells: transfection and viral transduction. AB - The corneal endothelium plays a key role in the physiology of the cornea, maintaining its transparency by regulating corneal hydration. Moreover, corneal endothelial cells play the central role in irreversible corneal graft rejection as human corneal endothelial cells are predominantly postmitotic, and destroyed cells cannot be replaced. Therefore, gene transfer to the corneal endothelium to modify the corneal immune response for prophylaxis of corneal endothelial rejection has become a fast-developing research field. An addition pivotal advantage of gene transfer to the cornea is the possibility of ex vivo transfection during organ culturing, minimizing the risk of systemic spread of the vector or the transgene expression. A wide variety of vectors has been found suitable for gene transfer to the corneal endothelium, and therapeutic efficacy has been demonstrated in some experimental models of corneal disease. However, the transfection efficiency varies widely among the different vectors, and the optimal transfection efficiency to provoke a desired effect is still unclear. Moreover, it certainly depends on the biological function of the chosen transgene (cytokine, growth factor, etc.). As a consequence, relatively few studies have been able to demonstrate significant prolongation of corneal allograft survival after gene transfer to the endothelium, and the ideal transfer strategy has not been found. In contrast, different transfer strategies compete today, each with its special advantages and disadvantages. Physical, viral, and nonviral techniques have been used to transfer transgenes into endothelial cells. In the introduction of this chapter, a short overview of the different gene transfer strategies for endothelial cells is given; the materials and methods sections describe in detail the most widely used viral gene transfer technique (adenoviral) and an important nonviral alternative technique (liposomal transfection) to endothelial cells. PMID- 19301675 TI - Creation of human skin equivalents for the in vitro study of angiogenesis in wound healing. AB - In our efforts aimed at studying the cellular responses to injury, including the angiogenesis of wound healing, we have developed a novel three-dimensional (3D) skin equivalent that is comprised of multiple cell types found in normal human skin or chronic wound beds. The in vitro model contains a microvascular component within the dermis-like extracellular matrix and possesses an intact epithelial covering comprised of skin-derived epithelial cells. Capillary endothelial cells can be labeled with fluorescent vital tracers prior to being embedded within a 3D matrix and overlaid with a monolayer of keratinocytes (normal or transformed). Once embedded in the matrix, the endothelial cells demonstrate capillary-like tube formation mimicking the microvasculature of true skin. Angiogenesis and the reepithelialization, which occur in response to injury and during wound healing, can be quantified using fluorescence-based and bright-field digital imaging microscopic, biochemical, or molecular approaches. PMID- 19301677 TI - Quantitative estimation of tissue blood flow rate. AB - Tissue blood flow rate (F) is a critical parameter for assessing functional efficiency of a blood vessel network following angiogenesis. This chapter aims to provide the principles behind estimation of F and a practical approach to its determination in laboratory animals using small, readily diffusible, and metabolically inert radiotracers. The methods described require relatively nonspecialized equipment. However, the analytical descriptions apply equally to complementary techniques involving sophisticated noninvasive imaging. Two techniques are described for the quantitative estimation of F using the tissue uptake following intravenous administration of radioactive iodoantipyrine (or other suitable radiotracer). The tissue equilibration technique is the classical approach, and the indicator fractionation technique, which is simpler to perform, is a practical alternative in many cases. The experimental procedures and analytical methods for both techniques are given, as well as guidelines for choosing the most appropriate method. PMID- 19301676 TI - Measurement of angiogenic phenotype by use of two-dimensional mesenteric angiogenesis assay. AB - Successful therapeutic angiogenesis requires an understanding of how the milieu of growth factors available combine to form a mature vascular bed. This requires a model in which multiple physiological and cell biological parameters can be identified. The adenoviral-mediated mesenteric angiogenesis assay as described here is ideal for that purpose. Adenoviruses expressing growth factors (vascular endothelial growth factor [VEGF] and angiopoietin 1 [Ang-1]) were injected into the mesenteric fat pad of adult male Wistar rats. The clear, thin, and relatively avascular mesenteric panel was used to measure increased vessel perfusion by intravital microscopy. In addition, high-powered microvessel analysis was carried out by immunostaining of features essential for the study of angiogenesis (endothelium, pericyte, smooth muscle cell area, and proliferation), allowing functional data to be obtained in conjunction with high-power microvessel ultrastructural analysis. A combination of individual growth factors resulted in a distinct vascular phenotype from either factor alone, with all treatments increasing the functional vessel area. VEGF produced shorter, narrow, highly branched, and sprouting vessels with normal pericyte coverage. Ang-1 induced broader, longer neovessels with no apparent increase in branching or sprouting. However, Ang-1-induced blood vessels displayed a significantly higher pericyte ensheathment. Combined treatment resulted in higher perfusion, larger and less branched vessels, with normal pericyte coverage, suggesting them to be more mature. This model can be used to show that Ang-1 and VEGF use different physiological mechanisms to enhance vascularisation of relatively avascular tissue. PMID- 19301678 TI - In vivo matrigel migration and angiogenesis assay. AB - The search for rapid and reproducible in vivo angiogenesis and antiangiogenesis assays is an area of intense interest. These types of assays are extremely useful in testing putative drugs and biological agents and for the comparison and enhancement of in vitro tests. The Matrigel plug assay is one such assay and has proved to be a relatively quick and easy method to evaluate both angiogenic and antiangiogenic compounds in vivo. Initial indications of the levels of activity of strong angiogenic or antiangiogenic compounds can be visually assessed even as the plugs come out of the mouse because there are color differences in the plugs compared to the controls. Further quantitation is then needed to determine levels of angiogenic/antiangiogenic activity, and this can be performed using a variety of methods. This chapter presents an overview of the basic methods used to set up both angiogenic and antiangiogenic assays, discusses factors influencing variability, and discusses the methods for quantitating the plugs obtained. The Matrigel plug assay provides another useful tool in angiogenesis research. PMID- 19301679 TI - The sponge implant model of angiogenesis. AB - The host response observed after the application of an appropriate stimulus, such as mechanical injury or injection of neoplastic or normal tissue implants, has allowed the cataloguing of a number of molecules and cells involved in the vascularization of normal repair or neoplastic tissue. Implantation of sponge matrices has been adopted as a model for the accurate quantification of angiogenic and fibrogenic responses as they may occur during wound healing in vivo. Such implants are particularly useful because they offer scope for modulating the environment within which angiogenesis occurs. A sponge implantation model has been optimised and adapted to characterise essential components and their roles in blood vessel formation in a variety of physiological and pathological conditions. As a direct consequence of advances in genetic manipulation, mouse models (i.e., knockouts, severe combined immunodeficient [SCID], nude) have provided resources to delineate the mechanisms regulating the healing associated with implants. Here, we outline the usefulness of the cannulated sponge implant model of angiogenesis and provide a detailed description of the methodology. PMID- 19301680 TI - The dorsal skinfold chamber: studying angiogenesis by intravital microscopy. AB - Intravital microscopy represents an internationally accepted and sophisticated experimental method to study angiogenesis, microcirculation, and many other parameters in a wide variety of neoplastic and nonneoplastic tissues. Since 1924, when the first transparent chamber model in animals was introduced, many other chamber models have been described in the literature for studying angiogenesis and microcirculation. Because angiogenesis is an active and dynamic process, one of the major strengths of chamber models is the possibility of monitoring angiogenesis in vivo continuously for up to several weeks with high spatial and temporal resolution. In addition, after the termination of experiments, tissue samples can be excised easily and further examined by various in vitro methods, such as histology, immunohistochemistry, and molecular biology. This chapter describes the protocol for the surgical preparation of a dorsal skinfold chamber in mice as well as the method to implant tumors in this chamber for further investigations of angiogenesis and other microcirculatory parameters. However, the application of the dorsal skinfold chamber model is not limited to the investigation of neoplastic tissues. To this end, the investigation of angiogenesis and other microcirculatory parameters of nonneoplastic tissues such as tendons, osteochondral grafts, or pancreatic islets have been objects of interest. PMID- 19301681 TI - The corneal pocket assay. AB - Continuous monitoring of neovascular growth in vivo is required for the development and evaluation of drugs acting as suppressors or stimulators of angiogenesis. The cornea assay consists of the placement of an angiogenesis stimulus (tumor tissue, cell suspension, growth factor) into a micropocket produced in the cornea thickness to evoke vascular outgrowth from the peripherally located limbal vasculature. Neovascular development and progression can be modified by the presence of locally released or applied inhibitory factors or by systemically given antiangiogenic drugs. This assay has the advantage over other in vivo assays of measuring new blood vessels only since the cornea is initially avascular. The experimental details of the avascular cornea assay and its advantages and disadvantages in different species are discussed. PMID- 19301682 TI - Use of the hollow fibre assay for studies of tumor neovasculature. AB - In vivo preclinical assays are required to screen potential agents that target the tumor vasculature. Here, a hollow fibre-based assay for the quantification of neovasculature in the presence or absence of an agent that potentially targets tumor neovasculature is described. The neovasculature is developed as a consequence of the presence of tumor cells encapsulated in hollow fibres, which are transplanted subcutaneously in the dorsal flanks of mice. PMID- 19301683 TI - The cranial bone window model: studying angiogenesis of primary and secondary bone tumors by intravital microscopy. AB - The successful treatment of primary and secondary bone tumors in a huge number of cases remains one of the major unsolved challenges in modern medicine. Malignant primary bone tumor growth predominantly occurs in younger people, whereas older people predominantly suffer from secondary bone tumors since up to 85% of the most frequently occurring malignant solid tumors, such as lung, mammary, and prostate carcinomas, metastasize into the bone. It is well known that a tumor's course may be altered by its surrounding tissue. For this reason, reported here is the protocol for the surgical preparation of a cranial bone window in mice as well as the method to implant tumors in this bone window for further investigations of angiogenesis and other microcirculatory parameters in orthotopically growing primary or secondary bone tumors using intravital microscopy. Intravital microscopy represents an internationally accepted and sophisticated experimental method to study angiogenesis, microcirculation, and many other parameters in a wide variety of neoplastic and nonneoplastic tissues. Since most physiologic and pathophysiologic processes are active and dynamic events, one of the major strengths of chronic animal models using intravital microscopy is the possibility of monitoring the regions of interest in vivo continuously up to several weeks with high spatial and temporal resolution. In addition, after the termination of experiments, tissue samples can be excised easily and further examined by various in vitro methods such as histology, immunohistochemistry, and molecular biology. PMID- 19301684 TI - Current approaches to the challenge of coronary heart disease risk assessment: C reactive protein mania and imaging-itis? PMID- 19301685 TI - Maintenance of cardiovascular risk goals in veterans with diabetes after discharge from a cardiovascular risk reduction clinic. AB - The authors evaluated maintenance of achieved cardiovascular risk control after discharge from a pharmacist-coordinated cardiovascular risk reduction clinic. Using data from 2001 to 2004 divided by financial quarters (ie, 3-month periods), the authors performed survival analysis of diabetic patients who had attained at least one cardiovascular risk goal in the clinic. Mean times to failure were 7.1 +/- 0.21 quarters for hemoglobin A1c, 7.6 +/- 0.29 quarters for low-density lipoprotein cholesterol (LDL-C), and 2.5 +/- 0.24 quarters for systolic blood pressure (SBP). Body mass index predicted glycemic control failure (hazard ratio [HR], 1.08; 95% confidence interval [CI], 1.01-1.15; P = .02), insulin use predicted LDL-C control failure (HR, 3.08; 95% CI, 1.15-8.22; P = .03), and baseline SBP predicted SBP control failure (HR, 1.02; 95% CI, 1.01-1.03; P = .0003). The authors found good durability of effect for most cardiovascular risk targets. Worse control at entry predicted failure after successful attainment of a cardiovascular goal. More sustained attention or booster interventions for patients with worse control at entry may be necessary. PMID- 19301687 TI - Association of total cholesterol/ high-density lipoprotein cholesterol ratio with proximal coronary atherosclerosis detected by multislice computed tomography. AB - The authors assessed the association between an elevated total cholesterol to high-density lipoprotein cholesterol (TC/HDL-C) ratio (> or = 4) and proximal coronary artery disease (CAD), as observed on multislice computed tomography. Coronary multislice computed tomographic angiography (96% on 40- or 64-slice) was performed in 295 individuals (39% women; mean age, 54 +/- 13 years) without documented CAD who were referred for coronary evaluation. Significant CAD was defined as > or = 50% stenosis in the left main, proximal left anterior descending, or > or = 2 epicardial vessels. Proximal plaque was defined as presence of any plaque in left main or proximal left anterior descending vessels. Individuals with an elevated TC/HDL-C ratio vs those without had a higher prevalence of proximal plaque (62% vs 48%, P = .04) and significant CAD (19% vs 9%, P = .009). On multivariate logistic regression analysis, only age, sex, and TC/HDL-C ratio > or = 4 were associated with significant CAD and proximal plaque. PMID- 19301686 TI - Control of lipids at baseline in the Bypass Angioplasty Revascularization Investigation 2 Diabetes (BARI 2D) trial. AB - In order to examine lipids, a major treatment parameter in those with diabetes and heart disease, the authors analyzed baseline data from the Bypass Angioplasty Revascularization Investigation 2 Diabetes (BARI 2D) trial. The study consisted of 2368 participants with type 2 diabetes and coronary artery disease from 49 sites in 6 countries (2295 provided lipid measurements). Fifty-nine percent of participants had a low-density lipoprotein (LDL) cholesterol level < 100 mg/dL. Levels of total, LDL, and non-high-density lipoprotein (HDL) cholesterol and triglycerides differed by age group (younger than 55, 55-64, and 65 years and older); they were lowest in those aged 65 years. Women had higher total, LDL, and non-HDL cholesterol values. Education was associated with lower total, LDL, and non-HDL cholesterol levels. LDL cholesterol and triglyceride values were lower in the United States and Canada. Adjustment for age, sex, education level, randomization year, and medication did not eliminate these differences. Geographic variation was seen and was not fully accounted for by demographic or treatment characteristics (all P values < .05). PMID- 19301688 TI - QT dispersion and heart rate predict the risk of sudden unexpected cardiac death in men: the Manitoba Follow-Up Study. AB - As many as half of all sudden cardiac deaths are unexpected, with no preceding symptoms or signs of cardiac problems. Since 1948, the Manitoba Follow-up Study has prospectively recorded routine medical information and resting electrocardiographic (ECG) findings from a cohort of 3983 men. During 58 years of follow-up, 180 men experienced sudden unexpected cardiac death (SUCD). Heart rate, the longest QT interval, the shortest QT interval, and their difference and QT dispersion (QTD) on ECGs recorded prior to SUCD and 5 years and 10 years earlier were compared with QT intervals on ECGs of age-matched controls. QTD and heart rate each were significantly (P < .01) and independently associated with increased risk for SUCD. Only primary prevention can reduce the risk for SUCD. Hence, this relationship between QTD and heart rate and SUCD emphasizes the importance of longitudinal noninvasive QT measurements on routine ECGs in healthy men. PMID- 19301689 TI - Use of carotid ultrasound to identify subclinical vascular disease and evaluate cardiovascular disease risk: summary and discussion of the American Society of Echocardiography consensus statement. PMID- 19301690 TI - Is there a need to continue anticoagulation following "successful" atrial fibrillation ablation? AB - Atrial fibrillation (AF) is the most common clinically significant arrhythmia worldwide, and its incidence is increasing. There has been increasing interest in ablation therapy to treat atrial fibrillation. One reason some patients undergo AF ablation might be to obviate the need for warfarin therapy, although current guidelines do not support this rationale. The current review shows that it is difficult to define a true "cure" postablation, as many of these patients will go on to experience future paroxysms of AF (either symptomatic or silent). The mechanism underlying embolism in patients with AF is not completely understood, and no long-term evidence exists that "successfully ablated" patients return to a baseline risk of stroke comparable to an AF-naive population. The authors recommend continued long-term anticoagulation post-AF ablation in patients satisfying CHADS criteria for elevated stroke risk. PMID- 19301691 TI - Ongoing Telmisartan Alone and in Combination With Ramipril Global Endpoint Trial (ONTARGET): implications for reduced cardiovascular risk. AB - The recently published Ongoing Telmisartan Alone and in Combination With Ramipril Global Endpoint Trial (ONTARGET) in patients with vascular disease or high-risk diabetes, as the largest published comparative trial of these agent classes, provides further evidence concerning the comparison between the angiotensin receptor blockers (ARBs) and the angiotensin-converting enzyme inhibitors (ACEIs). In this trial, telmisartan (an ARB) was non-inferior to ramipril (an ACEI) in reducing fatal and nonfatal cardiovascular events. Moreover, ONTARGET is an example of a high-quality noninferiority trial. However, the combination of the 2 agents was associated with more adverse effects without an increase in benefit. The study differed from several other comparative studies in which the dose and choice of ACEI was left to individual physicians. Further, in ONTARGET, the ACEI was not titrated to the maximum dose and patients with heart failure were excluded. PMID- 19301692 TI - Diabetes control and cardiovascular risk, Part II: Intensive glucose control- UKPDS follow-up. PMID- 19301693 TI - Lymphoscintigraphic evidence of lymph vessel dilation in the limbs of children with Brugia malayi infection. AB - Lymphatic filariasis (LF) is targeted for global elimination by the year 2020. It was earlier believed that LF is mostly a disease of adults. Recent studies indicate that in endemic countries filarial infection starts mostly in childhood even though the disease manifestations occur much later in life. The initial damage to the lymph vessels where the adult worms are lodged is dilation, thought to be irreversible even with treatment. Most of these studies relate to bancroftian filariasis. Studies that address this early pathology in brugian filariasis in humans are scarce. We report here for the first time, the lymphatic abnormalities seen on lymphoscintigraphy (LSG) in children with Brugia malayi filariasis. LSG was performed in 100 children aged between 3-15 years, who were enrolled in the study either because they were microfilaremic; had present or past filarial disease or were positive for antifilarial IgG4 antibodies. Inguinal and axillary lymph nodes were imaged in most children. Dilated lymph vessels were visualized in 80 children and this pathology was evenly distributed in all the three study groups. Lymph vessels dilation was seen even in three year old children. The implications of these findings for management of LF and control programmes are discussed. PMID- 19301694 TI - The plight of chronic filarial lymphoedema patients in choice of health care and health care providers in Pondicherry, India. AB - Lymphatic filariasis has a wide spectrum of clinical manifestations with asymptomatic parasite carriers at one end and irreversible lymphoedema of extremities at the other. Irreversible lymphoedema of extremities is one of the disabling conditions that drive the affected patients to seek treatment from various systems of medicines and health care providers. This study attempts to map the care seeking pattern and behaviour of patients with chronic filarial lymphoedema of lower limbs in an urban area. Consecutive filarial lymphoedema patients from the VCRC filariasis clinic were recruited for the study. A pre tested semi-structured questionnaire was used for interrogation of the patients. A total of 56 lymphoedema patients participated in the study. Majority (94.6%) of the patients sought medical management only. There was no difference (P>0.05) between the proportion of patients attending government (37.5%) and private (44.3%) medical care facilities There was also no difference in the proportion of patients' first consultations in private or government health care facilities. About 57.1% patients approaching governmental institutions opted for primary/secondary health care system. No particular sequential pattern of seeking health care was observed and the 56 study subjects followed 40 treatment-seeking routes by switching from one care provider to the other. The causes of not coming to the clinic for further check-up were 'no acute attacks' (30.4%), 'reduction in oedema volume' (21.7%), 'advised treatment being taken at home' (26.1%) and 'loss of daily wages' (21.7%). The study highlights the need to involve the private medical sector in morbidity management of filarial lymphoedema and to make governmental health facilities more accessible and user-friendly. PMID- 19301695 TI - Bacterial meningitis: rapid diagnosis and microbial profile: a multicentered study. AB - Meningitis continues to be a formidable illness with high morbidity and mortality among children in India. The present study was undertaken to prospectively look for the prevalence of pyogenic meningitis at Gulbarga and to find out the utility of gram stain, Latex Agglutination Test and (LTA) and C-reactive protein in the rapid diagnosis of pyogenic meningitis from children. Over a 48-months period, 535 children with a presumptive clinical diagnosis of acute bacterial meningitis were investigated by direct microscopy, CRP, bacterial culture, latex agglutination test (L TA), cell count and cell type and biochemical tests. Latex Agglutination Test (LA T) was done for detection of the antigens of Streptococcus pneumoniae, Group B Streptococci, E. coli, Neisseria meningitidis and Haemophilus influenzae type b. Among 535 suspected meningitis cases, 291 cases were diagnosed as pyogenic meningitis cases based on biochemical tests, cell count and cell type. Out of 291 cases, 55 cases have already received antibiotic treatment. Among 236 cases of untreated pyogenic meningitis cases, 199 CSF samples were culture positive. Streptococcus pneumoniae (44.7%) was the predominant organism identified, followed by H influenzae (25.6%) and Gp. B. Streptococci (9.5%). 208 of 236 cases were gram-stain positive, 129 cases had elevated CSF-CRP and 214 cases were diagnosed as pyogenic meningitis by the detection of bacterial antigens by latex agglutination test. Among 55 pretreated cases, only 05 (9.1%) CSF samples were culture positive, bacteria was observed in 36 gram stain smear, CRP was elevated in 16 CSF samples and 52 pretreated cases of suspected meningitis were diagnosed as pyogenic meningitis by latex agglutination test for detection of bacterial antigens. Many of the bacterial isolates were sensitive to gentamicin, cefotaxime and ceftriaxone and least sensitive to tetracycline and gentamicin. 13.1% of gram-negative bacilli were ESBL producers. To conclude, inclusion of latex agglutination test for detection of bacterial antigen in the routine diagnosis adds a valuable adjunct in the rapid and accurate diagnosis of pyogenic meningitis. PMID- 19301696 TI - In vivo antimalarial activity of leaves of Plectranthus amboinicus (lour) spreng on Plasmodium berghei yoelii. AB - An invivo study of aqueous extract of the leaves of Plectranthus amboinicus on Plasmodium berghei yoelii was conducted on laboratory infected albino mice and compared with standard drug chloroquine. Reduction of parasitemia at 250 mg/kg and 500 mg/kg of aqueous extract for 24 hrs, 48 hrs, 72 hrs and 96 hrs were determined. The reduction of parasitemia after 96 hrs was 100%, 67.9% and 76.2% for standard, 250 mg/kg and 500 mg/kg of aqueous extract respectively. The isolation of active principle responsible for the reduction of parasitemia may give a promising drug molecule. PMID- 19301697 TI - Studies on leptospirosis outbreaks in Peddamandem Mandal of Chittoor district, Andhra Pradesh. AB - An outbreak of leptospirosis in Peddamandem Mandal, Chittoor district, Andhra Pradesh occurred during Aug to Oct 2005. Out of 86 single human sera samples of suspected cases collected during the investigation, 49 (56.97%) samples from seven villages were found positive for leptospirosis both by DGM tests and IgM antibodies. Out of total 49 positive cases 16 (47.05%) were male and 33 (69.46%) female patients. The mean age of the positive cases were 42.7 years. There was no significant differences in male and female ratio and age groups in affected population. The higher degree of seropositivity was observed in adult females as they were mainly engaged in both domestic and peridomestic works. Geographical clustering of cases was evident. All the 49 positive cases had fever (100%). Myalgia (42.9%), stiffness of calf muscles (55.1%) and headache (32.6%) were the other major clinical features observed. There was only 1 (2.04%) case with conjunctival suffusion. None of the case presented with jaundice. All the suspected cases were tested negative for malaria, typhoid and dengue fever. In Peddamandem, contaminated water stagnation due to heavy rainfall and frequent contact of barefooted villagers with the infected sources registered higher incidence of leptospirosis. Most of the cases were from the contaminated water logged areas of the affected villages. In the affected villages none of the individual occupational category showed a significant association with seropositivity. It indicated that the transmission was from the common single category source in the villages i.e. contaminated stagnant water. The villagers living with livestocks and rodents were significantly associated with seropositivity. PMID- 19301698 TI - Dual culture method to determine the relationship of gut bacteria of sandfly (Phlebotomus argentipes) with promastigotes of Leishmania donovani. AB - A simple dual culture agar plating technique has been developed and evaluated for its efficiency in determining the relationship of gut bacteria of sandfly with Leishmania donovani promastigotes. There are about twenty morphologically distinct bacterial colonies have been isolated from the gut homogenate of Phlebotomus argentipes. In dual culture method, each bacterial isolate was inoculated in one half of the plate and the promastigotes of Leishmania was inculcated in the other half by streaking. After incubation, the type of association was determined based on the presence or absence of promastigotes colonies. The reliability of this method was compared with broth dilution method in 96 well plate. PMID- 19301699 TI - Prevalence of mosquito species in Chhura block of Raipur district of Chhattisgarh state. AB - Mosquitoes act as vectors for several diseases including malaria and filaria. In Indian scenario, Chhattisgarh is endemic for both the diseases. The present study on prevalence of mosquito population in block Chhura of Raipur district of Chhattisgarh state was undertaken between August 2002 and March 2004 to asses the distribution pattern of various species of mosquitoes and their seasonal variations. Indoor resting mosquitoes were collected from five villages by aspirator tube in the morning (06:00-08:00 hr) using torchlight. Adult mosquitoes were identified using standard morphological and pictorial keys. Altogether 11 species belonging to three genera viz., Anopheles (7 species), Armigeres (1 species) and Culex (3 species) were recorded during study period. Average man hour mosquito density was high in February 2003 (62.15) & March 2003 (69.35) and declined gradually till June 2003. From July 2003, it started rising again and reached a peak in August (94.10)-September (91.60) 2003. The vector species i.e. Anopheles culicifacies and Anopheles annularis were recorded high in rainy and winter seasons. PMID- 19301700 TI - Prevalence of human cysticercosis and taeniasis in rural Goa, India. AB - A cross sectional study among 450 individuals selected by strafified random sampling was carried out in rural Goa to find out the prevalence of cysticercosis and taeniasis, as well as to study the role of various factors associated with this diseases. The study participants were administered a pre-tested structured questionnaire and subsequently blood and stool samples were examined. SPSS software was used to analyze the data statistically. The sero-prevalence of cysterosis was 22.4%, which increased with age. Prevalence of taeniasis was 9.7% by stool examination. Individuals with taeniasis were thrice more likely to have cysticercosis; however no association between sero-positivity for cysterosis and pork consumption as well as religion could be established. The study confirmed a high sero-prevalence of cysticercosis in Goa underscoring the need to general awareness about good cooking habits and sanitation. PMID- 19301701 TI - Prevalence of anaemia and impact of interventions in school going adolescent girls of Haryana. PMID- 19301702 TI - Natural vertical transmission of dengue virus in peak summer collections of Aedes aegypti (Diptera: Culicidae) from urban areas of Jaipur (Rajasthan) and Delhi. PMID- 19301703 TI - Typhus fever in Pondicherry. PMID- 19301704 TI - Epidemiological, clinical and entomological observations on lymphatic filariasis in urban Puri, Orissa. PMID- 19301705 TI - Preliminary observation on Aedes aegypti survey during lean transmission season in sub Himalayan town of India. PMID- 19301706 TI - Small bowel responses to enteral honey and glutamine administration following massive small bowel resection in rabbit. AB - The trophic effects of honey and glutamine in the healing and adaptation of the small bowel following intestinal resection were studied in some Nigerian non descript breeds of rabbits. Nine rabbits of mixed sexes with mean body weight of 1.45 +/- 0.55 kg were used. They were randomised into three treatment groups following 50% small bowel resection. Group A rabbits were placed on oral honey treatment, Group B on oral glutamine and group C on normal saline (control). All groups exhibited signs of small bowel adaptation (glutamine(B) honey (A) control (C) at the end of the experiment (4 weeks) with oral glutamine showing the best overall effects on intestinal mucosal growth and adaptation evidenced by significant increase (P<0.05) in residual bowel length (37.3%), villi width (20.0%), crypt depth (113.3%) and a non significant increase in villus height (33.3%) and cellular mass (10.2%). Honey showed a better effect than control with a significant increase (P<0.05) in villi width (18.2%), crypts depth (66.7%) and cellular mass (33.9%) and a non significant increase in gross residual bowel length (24.6%), and villus height (30.5%). Our result shows that honey and glutamine have trophic effects on bowel mucosa healing and hyperplasia and have potential therapeutic effects on massive bowel resection in humans. PMID- 19301707 TI - Correlates of cardiac autonomic neuropathy in Nigerian patients with type 2 diabetes mellitus. AB - Cardiac Autonomic Neuropathy (CAN) contributes significantly to increased cardiovascular morbidity and mortality in type2 diabetic patients. This manuscript aims to study the prevalence of CAN in T2DM patients in our environment and identify factors that correlate with and predict development of CAN. This is with the intention to prevent and intervene in these factors. One hundred and eight (108) T2DM patients were screened and investigated by the standard six non-invasive tests including heart rate variability, QTc and Valsalva manouvre. Patients who had abnormality in more than 2 of the tests were adjudged to have CAN. They were statistically compared with those without CAN. Thirty- seven (34.2%) patients had CAN. The commonest abnormality was in heart rate response to standing (76.9%) There was no significant difference in the mean age and sex of those with CAN and those without. Though significantly more males had CAN, when sex was entered into the logistic regression it did not correlate with CAN. Poor short-term glycaemic control as demonstrated by fasting blood glucose (130.87 + 56.1 vs 94.24 + 48.9 mg%, p < 0.05); serum creatinine (1.23 + 0.52 vs 0.86 + 0.39 mg%, p<0.05), presence of hypertension and first degree family history were significantly more in patients with CAN. There was more than 2-fold increased risk of developing CAN when serum creatinine was greater than 1.5 mg% (OR 2.013, 95% CI 0.543-2.657) and when FBG was more than 120 mg% (OR 2.106, 95% CI 0.834-2.897). CAN is common in our patients with T2DM. It is more prevalent in those with hypertension, first degree family history and renal damage. Poor short-time glycaemic control was also a contributory factor. PMID- 19301708 TI - Relationship between balance performance in the elderly and some anthropometric variables. AB - Ability to maintain either static or dynamic balance has been found to be influenced by many factors such as height and weight in the elderly. The relationship between other anthropometric variables and balance performance among elderly Nigerians has not been widely studied. The aim of this study was to investigate the relationship between these other anthropometric variables and balance performance among old individuals aged >60 years in Ibadan, Nigeria. The study used the ex-post facto design and involved two hundred and three apparently healthy (103 males and 100 females) elderly participants with ages between 60 years and 74 years, selected using multiple step-wise sampling techniques from churches, mosques and market place within Ibadan. They were without history of neurological problem, postural hypotension, orthopeadic conditions or injury to the back and/or upper and lower extremities within the past one year. Selected anthropometric variables were measured, Sharpened Romberg Test (SRT) and Functional Reach Test (FRT) was used to assess static balance and dynamic balance respectively. All data were summarized using range, mean and standard deviation. Pearson's product moment correlation coefficient was used to determine the relationship between the physical characteristics, anthropometric variables and performance on each of the two balance tests. The results showed that there were low but significant positive correlations between performance on FRT and each of height, weight, trunk length, foot length, shoulder girth and hip girth. (p<0.05). There was low significant and positive correlation between SRT with eyes closed and arm length, foot length and shoulder girth. (p<0.05) and there was low but significant positive correlation between SRT with eyes opened and shoulder girth and foot length (P<0.05). Anthropometric variables affect balance performances in apparently healthy elderly. PMID- 19301709 TI - APOC impact assessment studies: baseline ophthalmological findings in Morogoro, Tanzania. AB - The goal of the African Programme for Onchocerciasis Control (APOC) is to eliminate Onchocerciasis as a disease of public Health significance and an important constraint to socio-economic development in the 19 none OCP (Onchocerciasis Control Project) countries covered through Community-Directed Treatment with Ivermectin, CDTI. In 1998, impact assessment studies were carried out in Morogoro, Tanzania during which baseline ophthalmological parameters were established. The hypothesis being tested is that CDTI will prevent or delay progression of onchocercal eye lesions and blindness. A total of 425 subjects aged 10 years or more from 14 villages within Bwakira district ofMorogoro region in Tanzania were examined for Snellen visual acuity, ocular microfilaria, lens opacities, uveitis and posterior segment disease especially chorioretinitis and optic nerve disease. Motion Sensitivity Screening Test (MSST) was carried out as well. Microfilaria was present in the anterior chamber of nearly half (49.2%) of all subjects examined. Prevalence of blindness was extremely high at 15.2%. Onchocercal lesions were responsible for blindness in 41.5% of these, followed by cataracts (27.7%), glaucoma (10.8%) and trachoma (6.2%). The main pathway to onchocercal blindness in this population was anterior uveitis with or without secondary cataracts. There is an urgent need to get CDTI underway and institute other horizontal primary eye care measures, especially cataract backlog reduction, in order to reduce the excessive burden of avoidable blindness in this community. PMID- 19301710 TI - Palmar and digital dermatogylyphic patterns of the three major ethnic groups in Nigeria. AB - Palmar and digital dermatoglyphic patterns of the three major ethnic groups in Nigeria were taken and their variability examined. Six hundred people were assessed, consisting of 200 Hausas (156 males, 44 females), 200 Ibos (98 males, 102 females), 200 Yorubas (112 males and 88 females). Clear prints were obtained by ink procedure and classified into patterns. It was found that ulnar loop was the most predominant among the ethnic groups. Comparison of digital and palmar ridge patterns among the ethnic groups revealed significant differences. The study established that digital and palmar ridge patterns vary significantly for the three ethnic groups. It was concluded from the study that dermatoglyphic traits of the digits and the palm may be used to differentiate the three ethnic groups. PMID- 19301711 TI - Antibiotic susceptibility patterns of Pseudomonas aeruginosa to available antipseudomonal drugs in Ibadan, Nigeria. AB - Pseudomonas aeruginosa has been reported to be a leading cause ofnosocomial infections. Resistance of this notorious bacterium to commonly used antimicrobial agents is becoming an increasing clinical problem and a recognized public health threat because there are limited number of antimicrobial agents including the antipseudomonal penicillins, cephalosporins, carbapenems, aminoglycosides and fluoroquinolones with reliable activity against it. This study was therefore carried out, using Bauer-Kirby method, to determine the antibiotic susceptibility patterns of Pseudomonas aeruginosa isolates from in-patients and out-patients attending the University College Hospital, Ibadan in Nigeria between June 2004 and May 2006. The isolation rate of Pseudomonas aeruginosa in clinical specimens was found to be 16.8% with the highest occurrence of 41.9% in ear swab followed by 39.3% occurrence in wound swab. The susceptibility pattern showed that 78.3% were sensitive to amikacin and 72.0% to ciprofloxacin. The isolates from the in patients showed higher resistance to all the antibiotics tested than the isolates from the out-patients, most especially amikacin and ciprofloxacin. However, no consistent antibiotic susceptibility pattern could be established for this pathogenic bacterium based on sources. In conclusion, the Pseudomonas aeruginosa species harboured by in-patients showed higher rates of antibiotic resistance than those of the out-patients. Also amikacin and ciprofloxacin were the two antibiotics found to be most potent against this pathogen. PMID- 19301712 TI - The relationship between central corneal thickness and intraocular pressure: a comparative study of normals and glaucoma subjects. AB - The purpose of the study is to determine whether central corneal thickness (CCT) is a better predictor than intraocular pressure (IOP) in early identification of those at higher risk of developing glaucoma. Sixty-five subjects were categorized into normals, ocular hypertensives and glaucoma subjects based on clinical characteristics of ocular risk factors. The lOP was assessed with slit-lamp mounted Goldmann applanation tonometer. Prior to applanation tonometry, the central corneal thickness (CCT) of both eyes was assessed with Sonomed PacScan 300AP Biometric/pachymeter. The difference in mean IOP between normals and glaucoma subjects was statistically significant (unpaired t-test; p<0.05). Similarly, there was a significant difference in mean CCT between normals and glaucoma subjects (p<0.05). The association between CCT and Age was not significant in normals but slightly significant in glaucoma subjects and the linear regression predicts a decrease of 7.0 im in CCT for every 10 years. A strong association was found between CCT and IOP for ocular hypertensives with a prediction of increase of 0.70 mmHg for every 10 im corneal thickening. The association between CCT and IOP for glaucoma subjects was weak, with an indication of an increase of 0.35 mmHg in intraocular pressure for every 10 im corneal thinning. The central corneal thickness is a better predictor than intraocular pressure in identifying those at higher risk of developing primary open-angle glaucoma when combined with some ocular risk factors. PMID- 19301713 TI - Challenges of glaucoma service delivery in Federal Medical Centre, Azare, Nigeria. AB - To find out the challenges of management of glaucoma, an assessment of glaucoma services at the Federal Medical Centre, Azare was carried out through oral interviews of ophthalmic staff, enquiring about available personnel and diagnostic equipment from relevant sectors of the Hospital and local authorities. A retrospective analysis of records of glaucoma patients (aged > 20 years) who presented at the centre between February 2002 and July 2007 was carried out. The ophthalmology unit has 6 ophthalmic nurses, 1 ophthalmologist and 1 optometrist. Instruments available were: A Schiotz tonometer, a slit lamp biomicroscope, a Volk +90 Diopter lens, a Goldman applanation tonometer, a l-mirror gonioscopy lens, a non contact applantation tonometer and a visual field analyzer. All the patients treated conservatively had guttae timolol 0.5% either solely (64 eyes, 50.8%) or in combination with other intraocular pressure lowering eye drops. Only one patient was treated with guttae latanoprost 0.005% (Xalatan). The only type of surgery carried out was Trabeculectomy in 22 eyes (14.0%). No intra or post operative antifibrotic agent was used. There is a dire need of human and infrastructural facilities for adequate management of glaucoma in the centre. PMID- 19301714 TI - Prevalence of premenstrual syndrome and its relationship with blood pressure in young adult females. AB - Premenstrual syndrome is a collection of symptoms which women experience 1 to 2 weeks before menstruation which clear with onset of menstrual flow and capable of interfering with life functions. An association between Premenstrual syndrome and hypertension has been suggested leading to the suspicion that it may be one precussor trait of hypertension in the normal population. Since people who develop hypertension later in life start from the higher range of normal earlier on, we sought to find if premenstrual syndrome had any bearing on hypertension. Consenting female subjects in three institutions in Jos, Nigeria were studied. Self-administered questionnaires designed to diagnose premenstrual syndrome and anxio-depressive status were used. Medical, family and menstrual history, as well as height, weight, pulse and blood pressures were documented; and data analysed. Four hundred and fourty seven subjects aged between 17 and 38 years with a mean of 23.6 + 3.9 were studied. Sixty one percent of them satisfied the criteria for diagnosis of premenstrual syndrome. When the data for those in luteal phase was split into the early (1st) and late (2nd) parts, there was a statistically significant rise in systolic and diastolic blood pressure from 1st to 2nd parts only for those with premenstrual syndrome (SBP 107.3 + 11.05 to 111.39 + 13.48 mm Hg. p=0.05 and DBP 65.06 + 10.38 to 70.69 + 10.03. p=0.004). Those with premenstrual syndrome also had higher anxiety and depression scores (chi-square 47.9 and 28.4 respectively). The symptoms of premenstrual syndrome peak in the 2nd part of the luteal phase. They are associated with a lot of stress. Since blood pressure elevation with this change was significantly higher among subjects with premenstrual syndrome, it is felt that premenstrual syndrome may predict future hypertension among currently normotensive females. PMID- 19301715 TI - Histopathological analysis of non-squamous cell malignancies of the uterine cervix in Maiduguri, Nigeria. AB - Carcinoma of the uterine cervix is the most common gynaecological malignancy in the developing world despite being largely preventable. The non-squamous cancers of the cervix have not been properly documented in our setting. This study is aimed at reviewing the histological types ofnon-squamous cell cancers of the uterine cervix in relation to the age of presentation, and to analyze some of the histopathological features of adenocarcinomas. Cases of cancer of the cervix histologically diagnosed between January 1989 and December 2004 were extracted from the cancer register of the University of Maiduguri Teaching Hospital. The request forms, slides, and some tissue blocks were retrieved from the archives of the Histopathology Department. The cases were histologically reviewed and the data analyzed using simple statistical methods. A total of 491 cases of cervical cancers were diagnosed. Of these, 432(88.0%) cases were squamous cell carcinomas while 59(12.0%) were non-squamous cell malignancies. Out of the 59 non-squamous cervical cancers, 42(71.2%) were endocervical-type adenocarcinomas, while serous papillary, clear cell, and adenosquamous carcinomas comprised 2(3.4%) cases each. Seven (11.9%) cases were metastatic choriocarcinomas. Over 60% of these cases occurred within the third and fourth decades of life. Adenocarcinomas are the most common non-squamous malignancies of the cervix and their age of presentation was similar to that of their squamous cell counterpart. Haemorrhage and lymphocytic infiltrates were significantly associated with tumour grade while necrosis was independent of tumour grade. PMID- 19301716 TI - Pattern and outcome of medical admission of elderly patients seen at University of Ilorin Teaching Hospital, Ilorin. AB - The elderly are increasingly been hospitalised into medical wards in Nigeria and little information is currently available on this group of patients. The objective of this study was to describe the demographic, clinical characteristics and outcome of management of elderly patients admitted into medical wards at the University of Ilorin Teaching Hospital Ilorin, Nigeria. A retrospective study in which hospital records of patients admitted between years 2001 to 2004 were reviewed. Information obtained included gender, occupation, diagnosis, investigations, duration of stay and outcome management. A total of 4113 adults were admitted into the medical wards within the period under review. Of these, 456 were aged 60 years and above. The elderly patients accounted for 11.1% of total hospitalisation into the medical wards. The mean age of the patients was 69 +/- 9 years with male:female ratio of 243:105. The 3 most common diagnoses were: hypertensive heart failure (HHF), 19%; cerebrovascular accident (CVA), 12%; and tuberculosis (TB), 11%. The mean duration of hospitalisation was 15.6 +/- 13.8 days. Cases of mortality had significantly higher value of mean serum potassium, urea and creatinine, compared to those with favourable outcome and were eventually discharged home. A total of 192 patients (55.2%) were discharged home, while 109 (31%) died giving in-hospital mortality of 31.7%. Majority of the deaths (75%) occurred within 14 days of hospitalisation. Significantly higher number of patients died within the first seven days compared to those discharged (P<0.01). The mean duration of hospital stay was 15.6 +/- 13.8 days. Patients with favourable outcome spent a mean of 18.5 +/- 14.1 days while cases of mortality had a mean of 10.4 +/- 8.8 days in hospital before death. Geriatric patients constituted more than a tenth of total hospitalisation into UITH medical wards. They accounted for a significant proportion of in-hospital mortality. Since a third of the death occurred within the first few days of admission, improvement in the management of acute medical cases especially in the elderly is urgent needed. This will ensure survival of greater number of patients and thus reduces mortality. PMID- 19301717 TI - Epidemiological patterns of head injury in a newly established neurosurgical service: one-year prospective study. AB - Head injury is a disease afflicting mainly young males, and road traffic accident is the most common aetiologic factor. This report evaluates the findings in a one year prospective study done from April 21, 2006-April 20, 2007 in the first year of services in one of Nigeria's new neurosurgical Centres, to establish the baseline epidemiological patterns of head injury. Data was collected using a questionnaire from the point of presentation till discharge of each head injury patient, and augmented with theatre and outpatient records, and analyzed. The Glasgow Coma and Outcome Scales were used for grading. Our Centre is a tertiary health facility that receives referrals from private, primary and secondary facilities. Of the 334 total patients treated, 210 (62.9%) had head injuries. Males were 158 (75.2%), and 145 (69%) resulted from road traffic accident, mostly from motorcycles. Fractures were mostly basal 86 (41%), brain pathology was mostly cerebral contusion 74 (35.2%), and treatment mostly nonoperative 137 (65.2%), with good outcome in 144(68.6%). Mortality was 40 (19.1%). Trauma is the main reason for neurosurgical consultation in our Centre and the unsafe use of roads, especially with motorcycles, remains the major cause of head injury. PMID- 19301718 TI - A three year review of sciatic nerve injection palsy in the Physiotherapy Department of a Nigerian Specialist Hospital. AB - Sciatic Nerve palsy associated with intramuscular injection is a major cause of disability among children under 6-years-old in the developing Countries. A retrospective study was conducted with an objective to review cases of all patients with Injection induced sciatic nerve palsy managed at the Physiotherapy Department, State Specialist Hospital, Akure Ondo State over a period of 3 years (January 2004 and December 2006). The case notes of these patients were reviewed and the details of the patients' sex, age and presentations were obtained. Records of 160 patients presenting with Sciatic Nerve Injection Palsy were studied. Males accounted for 60% while 40% were females. They were aged from 3 months to 70 years with paediatric cases accounting for 90% of cases. All of the patients had intramuscular injections following febrile illness. Muscle paralysis with foot drop accounted for 41% cases while 59% had varying degrees of muscle paresis, without foot drop, and 4% had equinovarus deformity as a complication of the paralytic foot drop. Sciatic Nerve Injection Palsy especially in children is a common referral to the Physiotherapy Clinic. There is therefore need for caution in the administration of gluteal intramuscular injections particularly in children. PMID- 19301719 TI - An unusual case of total maxillary sequestration in a diabetic patient. AB - Osteomyelitis of the jaw is of relatively low incidence. Maxillary Osteomyelitis is however rare compared with mandibular osteomyelitis. The extensive blood supply and the strut-like bones of the maxilla make it less prone to chronic infections. Systemic conditions that alter the host's resistance such as diabetes mellitus, autoimmune disorders, agranulocytosis, anaemia, especially sickle cell anaemia are predisposing factors for the development of this condition. An unusual case of chronic maxillary osteomyelitis induced by trauma in a diabetic, with subsequent atypical necrotising ulceration of palatal mucosa resulting in total maxillary sequestration is presented. PMID- 19301720 TI - Efficacy of celecoxib on controlling irregular uterine bleeding secondary to Jadelle use. AB - OBJECTIVE: To evaluate the efficacy of celecoxib and placebo for controlling irregular uterine bleeding in Jadelle users. DESIGN: Randomized double blind placebo controlled trial. SETTING: Family Planning Clinic, King Chulalongkorn Memorial Hospital, Bangkok, Thailand. MATERIAL AND METHOD: Forty Jadelle users with irregular bleeding were randomly allocated into two groups. Twenty users received celecoxib 200 mg once a day for five days and the other twenty users received the placebo in the same manner The participants were requested to maintain their daily record of bleeding, adverse effects, and satisfaction. RESULTS: The percentage of the subjects whom bleeding was stopped within 7 days after initial treatment was significantly higher in the celecoxib group than in the placebo group (70% vs. 0%; p < 0.001). The mean duration of bleeding-free interval was significantly longer in celecoxib than placebo group (24.0 +/- 1.65 days vs. 10.0 +/- 6.50 days; p < 0.001). The mean duration of bleeding days was significantly shorter in celecoxib than placebo group (5.0 +/- 1.65 vs. 19.0 +/- 6.50 days; p < 0.001). Patients satisfaction in celecoxib group was significantly higher than the placebo group (80% vs. 30%; p < 0.001). There was no detectable adverse effect in both groups. CONCLUSIONS: Celecoxib was more effective than placebo in the short-term control of irregular bleeding in Jadelle users. The mechanism of nonsteroidal anti-inflammatory drugs (NSAIDs) for the reduction of endometrial bleeding is likely from COX-2 inhibition. PMID- 19301721 TI - Human telomerase reverse transcriptase (hTERT) expression in borderline ovarian tumors: an immunohistochemical study. AB - OBJECTIVE: To investigate the expression of human telomerase reverse transcriptase (hTERT) in epithelial borderline ovarian tumor (BOT) by immunohistochemistry with correlation to clinicopathologic variables. MATERIAL AND METHOD: Paraffin-embedded tissue sections of 62 borderline ovarian tumors (47 mucinous, 14 serous, and 1 clear cell) and 12 epthelial ovarian carcinomas were immunostained with antibodies to hTERT. The intensity and quantity of the immunostaining was determined and analyzed with clinicopathological characteristics. RESULTS: hTERT expression was detected in 48.4% of BOT and all cases of epithelial ovarian carcinoma. In immunoreactive BOT 50% of cases were scored as high expression. Serous BOT had the highest rate of hTERT expression. There was no significant statistical difference of hTERT immunoreactivity between histologic types of BOT. No hTERT immunoreactivity was observed in the benign parts of the same slides of each immunoreactive case. hTERT immunoreactivity was positively correlated with FIGO stage (p = 0.04), but not with other variables. The mean follow-up time of BOT cases was 81.63 months and no recurrence or death was noted. CONCLUSION: hTERT expression was found in half of BOT and all of epithelial ovarian carcinoma. High hTERT expression was associated with FIGO stage. PMID- 19301722 TI - Tetanus: a retrospective study of clinical presentations and outcomes in a medical teaching hospital. AB - BACKGROUND: Tetanus has been a serious infectious disease with high mortality and morbidity. Some clinical factors can predict its severity. OBJECTIVE: Survey the clinical profiles, their correlation with the severity of tetanus and the final outcomes after treatment. MATERIAL AND METHOD: A retrospective study of the medical records of patients with tetanus who attended Songklanagarind hospital between January 1982 and June 2008 was done. The correlation between clinical profiles and severity was analyzed by Fisher exact test to detect the significant correlation (p < 0.05). RESULTS: Forty-seven cases of tetanus, composed of 36 male and 11 female patients, were reviewed. The mean age of onset was 45.5 years (SD = 19.6 years). Generalized tetanus was the most common form found (91.4%). History of injury or wound was reported in 80.8%. Age over sixty, puncture wound, onset time of less than seven days, and generalized tetanus, significantly correlated with clinical severity. Only two cases died, but the rest of the cases had residual muscle rigidity after treatment. CONCLUSION: The overall clinical features of tetanus and their correlation with the disease severity were presented. Unlike in the past, patients with tetanus can now be reversible under appropriate intensive critical care. PMID- 19301723 TI - Enhancing limb salvage by non-mobilized peripheral blood angiogenic cell precursors therapy in patients with critical limb ischemia. AB - BACKGROUND: Stem cell therapy has been proposed to enhance the salvage of critically ischemic limbs. OBJECTIVE: Assess the efficacy and safety of the implantation of non-mobilized peripheral blood angiogenic cell precursors (NMPB ACPs) in patients with critical limb ischemia (CLI) who were poor candidates for standard revascularization treatment options. MATERIAL AND METHOD: Six patients with CLI due to the infrapopliteal artery occlusive disease were included in the present study. Intramuscular injections of NMPB-ACPs were administered in the ischemic limbs. The efficacy was evaluated by clinical outcomes, ankle brachial index, toe brachial index, and computerized tomographic angiography. RESULTS: There was no evidence of local or systemic complication related to the procedure. Five patients (83.3%) had clinically significant improvement of adequate circulation at the distal limb for the complete healing. Four of them had complete healing of ischemic ulcers and stumps of toe amputation. However one patient with adequate granulation tissue at the stump of the left first toe amputation subsequently suffered from severe foot infection originating from the other toes and eventually underwent below knee amputation. There was no improvement of circulation at the distal limb after the administration of NMPB ACPs in one patient (16.7%) who eventually underwent major amputation. CONCLUSION: The preliminary result of NMPB-ACPs therapy may be safe and provide benefits in the improvement of circulation in patients with CLI. A larger controlled trial is required to ascertain these preliminary results. PMID- 19301724 TI - A comparison of continuous femoral nerve block (CFNB) and continuous epidural infusion (CEI) in postoperative analgesia and knee rehabilitation after total knee arthroplasty (TKA). AB - BACKGROUND: Postoperative epidural analgesia (EA) and femoral nerve block (FNB) provided effective pain relief However, EA has common side effects such as nausea, vomiting, pruritus, dizziness, and hypotension. Some investigations found that those side effects were less in FNB than in EA. However the analgesic equivalent of both techniques have not been confirmed. OBJECTIVE: The authors compared continuous epidural infusion (CEI) with continuous femoral nerve block (CFNB) regarding the postoperative analgesic efficacy, side effects, postoperative knee rehabilitation, and hospital length of stay (LOS). MATERIAL AND METHOD: In this prospective, randomized controlled study, 61 ASA physical status I-III patients scheduled for elective unilateral total knee arthroplasty (TKA) under spinal anesthesia (SA) participated. The patients were allocated into two groups. In the ward, patients in Group I (CEI) were maintained by continuous infusion of 0.125% levobupivacaine with morphine 0.0125 mg/ml (4 ml/hr), Group II (CFNB) were maintained by 0.125% levobupivacaine (8 ml/hr). RESULTS: Patients in the CFNB group, the VAS scores at PO6-12 hr and tramadol IV requirement were significantly greater than the CEI group (VAS: PO6 hr p-value = 0.001, PO12 hr p value = 0.004). Patients in the CEI group experienced dizziness, pruritus, and PONV more than the CFNB group significantly. Patient satisfaction was greater with the CFNB group although postoperative knee rehabilitation and the hospital LOS were not different. CONCLUSION: CFNB represents the optimal analgesia with fewer side effects and greater patient, satisfaction. The rehabilitation indices and duration of hospital stay are comparable in both groups. PMID- 19301725 TI - The Thai Anesthesia Incident Monitoring study (Thai AIMS): an analysis of 21 awareness events. AB - OBJECTIVE: To demonstrate the characteristics, outcomes, and the circumstances associated with intraoperative recall of awareness. MATERIAL AND METHOD: Relevant data of intra-operative recall of awareness were extracted from the Thai Anesthesia Incident Monitoring study (Thai AIMS) database of 1996 incident reports and 2537 incidents which were conducted among 51 hospitals throughout Thailand from January to June, 2007. Details regarding patients, surgical, anesthetic and systematic factors were recorded in a structured data record form. The completed record forms were reviewed independently by three anesthesiologists. The descriptive statistic was analyzed by using SPSS software version 11.5 and demonstrated in number and percent. RESULTS: Twenty-one incidents (21/1996 = 1.05%) of intra-operative recall of awareness were reported. Awareness was predominantly found in females (76.2%) and with ASA physical status I (47.6%). Most of the patients recalled events during the maintenance period and reported sound (71.4%), pain (52.4%), feeling operated (38.1%), paralysis (33.3%), recognizing intubated (4.8%) and panic (4.8%). Anxiety (33.3%), temporary emotional stress (19%), and post traumatic stress (4.8%) were found during immediate outcome assessment but scarcely sustained on the hospital discharged date. The factors associated with the incidents were anesthetic related in the majority especially ineffective monitoring (100%), pre-medication abandonment (100%) and light anesthesia (71.5%). CONCLUSION: Intra-operative recall of awareness in the Thai AIMS was 1.05% of all incident reports. Most of the events were considered as anesthesia related. The suggested corrective strategies were quality assurance activity, effective monitoring and equipment maintenance. PMID- 19301726 TI - The Thai Anesthesia Incident Monitoring study (Thai AIMS): perioperative arrhythmia. AB - BACKGROUND: The Royal College of Anesthesiologists of Thailand organized the first national sentinel incident reports of anesthesia related adverse events in 2007 on an anonymous and voluntary basis. The aims of the present study were to analyze incidence, risk factors, clinical course and outcome of perioperative arrhythmia and indicate the contributing factors and suggested corrective strategies in the database of the Thai Anesthesia Incidents Monitoring Study (Thai AIMS). MATERIAL AND METHOD: This study was a prospective descriptive multicentered study conducted between January 2007 and June 2007. Data was collected from 51 hospitals across Thailand. All cases whose arrhythmia was detected intra-operatively and within 24 hr postoperative period were analyzed by 3 independents anesthesiologists. Any disagreements were discussed to achieve a consensus. RESULTS: Four hundred and eighty-nine cases were enrolled as relevant arrhythmia cases. Bradycardia was the most common type (434 cases; 88.8%). Most of all events occurred intra-operatively (94.7%) and electrocardiography was the most common firstly detected monitoring equipment (95.5%). Arrhythmia occurred frequently in patients with hypertension and pre-operative heart rate < 60 beat per min. Intravenous anesthetics, central neural blockage and vagal reflex were considered to be the 3 most common suspected causes of arrhythmia requiring treatment. Most common outcomes were minor physiologic change with complete recovery physiologic change with complete recovery while 7% of incidents developed fatal outcome. The most common contributing factor was human factor (72.4%) especially in experience. An experienced anesthetic team with high awareness could be the minimizing factors. CONCLUSION: Arrhythmia accounted for 19.2% of 2,537 incidents of the Thai AIMS database. Bradycardia was the most common type of cardiac arrhythmia. Most arrhythmia was benign but might be fatal. Suggested corrective strategies such as guidelines practice, improvement of supervision and quality assurance activity. PMID- 19301727 TI - Prevention of recurrent wheezing in young children by loratadine compared with ketotifen. AB - BACKGROUND: Various trials showed benefit of the prophylactic agent ketotifen in prevention of recurrent wheezing in young children, but no such clinical trial with loratadine or comparison trial is available. OBJECTIVE: To study the efficacy and safety of loratadine syrup compared with ketotifen and placebo in prevention of recurrent wheezing in young children. MATERIAL AND METHOD: Randomized double-blind placebo controlled trial on 90 recurrent wheezing children aged less than 6 years old was done. Children were randomized to receive loratadine, ketotifen syrup, or placebo with dose of 0.25 cc/kg once a day for four months. Blood biochemistry (CBC, LFT) and EKG were performed pre and post treatment period. Assessment of symptoms--wheezing and night cough including use of bronchodilators was done daily via patient diary card. Subjects were asked to do monthly visits to the clinic for physical examination. At those visits, the doctors questioned the patients about adverse event. RESULT: Of the 90 children enrolled, 12 dropped out. Thus, 27 children remained in the loratadine, 26 in the placebo, and 25 in the ketotifen group. The demographic data were comparable among the three treatment groups. It was noted that wheezing decreased significantly at 2 months in the ketotifen (p = 0.008) and at 3 months in the loratadine (p = 0.029) but not in the placebo group. Coughing at night decreased significantly at 3 months in both the loratadine (p = 0.005) and the ketotifen (p = 0.036) group. The use of bronchodilator drug was significantly decreased at 2 months in the ketotifen (p = 0.028) and placebo (p = 0.025) group, and at 3 months in the loratadine (p = 0.009) group. Only a few patients had mild adverse events in all groups. CONCLUSION: Loratadine and ketotifen are safe and effective significantly in prevention of recurrent wheezing in young children. PMID- 19301728 TI - Unrecognized congenital heart disease among Thai children. AB - OBJECTIVE: To report on the prevalence of unrecognized congenital heart disease (CHD) among elementary school children (231 schools, 6 cities in Tak province). MATERIAL AND METHOD: Between January, 2006 and December 2006, 38,055 children were examined by trained nurses and health officers to ausculate the precordium. Those with abnormal heart sounds were re-evaluated by pediatric cardiologists. RESULTS: Of the 278 subjects with abnormal heart sounds, 43 had proven heart disease, 40 showed CHD (1.05 per 1,000) and three had rheumatic valvular diseases. CONCLUSION: Understanding the case rate helped approximate the number of children who are currently undetected but have heart problems or may develop heart problems in the future. Every diseased person will receive treatment. PMID- 19301729 TI - Clinical features, management and outcomes of progressive outer retinal necrosis (PORN) in southern Thailand. AB - OBJECTIVE: To study the demographics, clinical features, treatment, and visual outcomes of progressive outer retinal necrosis (PORN) in a group of Thai patients. MATERIAL AND METHOD: All cases of AIDS with a clinical diagnosis of PORN in a major tertiary referral hospital in southern Thailand between January 2003 and June 2007 were retrospectively reviewed. Demographic data, clinical features, treatment regimens, and visual outcomes were analyzed. RESULTS: Seven patients (11 eyes) were studied. The mean age was 44.7 years. The median CD4 count was 12 cells/mm3. A known history of cutaneous zoster was documented in 57% of cases. The median follow-up period was 17 weeks. Fifty-seven percent of the patients had bilateral disease. A majority of eyes (45.4%) had initial visual acuity of less than 20/50 to equal to or better than 20/200. About two-thirds of the eyes had anterior chamber cells. Vitritis and retinal lesions scattered throughout both posterior pole and peripheral retina were found in 72.7%. Either intravenous acyclovir in combination with intravitreal ganciclovir injections or intravenous aclyclovir alone was used for initial treatment. Retinal detachment occurred in 54.5%. Final visual acuity worsened (loss of 3 lines on the ETDRS chart or more) in 60%. Visual acuity was no light perception in 45.5% at the final recorded follow-up. CONCLUSION: Demographics, clinical features and treatment outcomes of PORN in this group of Thai patients were comparable with studies from other countries. Visual prognosis is still poor with current treatment regimens. PMID- 19301730 TI - Photodynamic therapy for AMD and non-AMD patients: two-year results in Thais. AB - OBJECTIVE: To determine the long-term effect of photodynamic therapy (PDT) with verteporfin for age-related macular degeneration (AMD) and non-AMD in Thailand and to compare with the Treatment of Age-Related Macular Degeneration with Photodynamic Therapy (TAP) and Verteporfin in Photodynamic Therapy (VIP) study. MATERIAL AND METHOD: The data of patients who received PDT between July 2003 and December 2004 and had completed two-year follow-up were prospectively reviewed. Treated eyes were classified into two main groups, the AMD (group 1) and non-AMD (group 2) groups. The AMD group was further divided into three subgroups, group 1A, AMD with subfoveal choroidal neovascularization (CNV) and TAP/VIP compatible if they followed the recommendation guidelines characteristics, group 1B, AMD with subfoveal CNV and TAP/VIP incompatible, and group 1C, AMD with non-subfoveal CNV. The main outcomes were visual acuity change, number of treatments and the comparison with the first year results. RESULTS: Of 56 eyes, 46 eyes (82.14%) had completed 24-month follow-up. Thirty-four eyes had CNV-related AMD and 12 eyes were non-AMD. At the 24-month follow-up, mean visual acuity change in group 1A, 1B, 1C were increased 0.25 (p = 0.13), 0.05 (p = 0.52), and 0.28 (p = 0.003), respectively. The total number of treatments in the first and second year was 1.8 and 0.1 in group 1A, 2.3 and 0.1 in group 1B, 1.5 and 0.25 in group 1C. CONCLUSION: PDT was effective in Thai patients for the two-year follow-up even if they were not compatible with TAP/VIP criteria. The treatment demonstrated stabilization or less visual loss in long-term results. PMID- 19301731 TI - Unilateral vertical nystagmus: the Heimann-Bielschowsky phenomenon. AB - OBJECTIVE: To present clinical feature of Heimann-Bielschowsky phenomenon and strabismus association. DESIGN: Retrospective case series. MATERIAL AND METHOD: Medical records of cases between January 2004 and January 2008 were reviewed. Eight cases met the criteria. Age, sex, visual acuity, and eye examination data were collected. RESULTS: Six cases were male and two cases were female. Age at presentation ranged from 4 to 45 years old. Seven cases had vision of 6/60 or worse. Only one case had visual acuity 6/36. Strabismus was seen in five cases (exotropia 4 cases and esotropia 1 case). No one had symptoms of diplopia or oscillopsia. All cases had unilateral coarse, slow, pendular vertical oscillations occurring in the poor vision eye. CONCLUSION: Monocular vertical oscillation may develop in an eye with reduced vision. It may be a common condition but under diagnosed. PMID- 19301732 TI - Key nasal symptoms predicting a positive skin test in allergic rhinitis and patient characteristics according to ARIA classification. AB - OBJECTIVE: To find predicting symptom(s) in patients with a positive skin test, and identify patient characteristics according to ARIA classification. MATERIAL AND METHOD: Four hundred and thirty four rhinologic patients were retrospectively studied. Nasal obstruction, rhinorrhea, itching, sneezing, and smell dysfunction were assessed. Patients were also classified according to the ARIA guideline. RESULTS: Of 434 patients, 277 (63.8%) were skin prick test positive. There was no statistical difference in sex, but the mean age of the positive skin test group was lower than that in the negative group (p < 0.05). Intermittent or persistent symptoms and total symptom score were not significantly different. Severe nasal itching was more common in the positive group (p = 0.04). The impact of symptoms was similar between the two groups, except for self-reported comorbidity, which was higher in the negative group (p = 0.039). CONCLUSION: There were no predicting symptoms and no difference in the symptom characteristics for the patients with positive skin test. PMID- 19301733 TI - Comparison of conventional external radiotherapy and stereotactic radiotherapy in the treatment of pituitary adenoma. AB - OBJECTIVE: To compare the results of pituitary adenoma treated with conventional external beam radiotherapy (EBRT) versus stereotactic radiosurgery and stereotactic radiotherapy (SRS/SRT). METHOD AND MATERIAL: Data of patients with pituitary adenoma treated at Radiotherapy and Oncology unit, Ramathibodi Hospital between 1990 from 2003 were retrospectively collected and analyzed. Twenty-two patients were treated with EBRT and 51 patients were treated with SRS/SRT. RESULTS: The 5-year overall survival and local control rates were not different between the EBRT and SRS/SRT group (91% vs 100%, p = 0.10; 95% vs 96%, p = 0.33). The 5-year freedom from newly initiated hormonal replacement was 50% in EBRT and 75% in SRS/SRT group (p = 0.38). CONCLUSION: At a similar outcome but with lower expense of resources, EBRT should be considered an acceptable radiation technique for patients with pituitary adenoma, especially in developing countries such as Thailand. PMID- 19301734 TI - Late effects and cosmetic results of simultaneous integrated boost versus sequential boost after conventional irradiation in breast-conserving therapy; out come of 7 months follow-up. AB - OBJECTIVE: To compare simultaneous integrated boost and sequential boost after conventional irradiation in breast-conserving therapy in aspect of late effects and cosmetic results. MATERIAL AND METHOD: Between August 2006 and June 2007, 60 breasts were treated in this prospective nonrandomized study, designed to compare simultaneous integrated boost (additional 10 Gy/25F) and sequential boost (15 Gy/5F) to the tumor bed in terms of late effects and cosmetic results at 7-month and 3-year follow-up. Pearson Chi-square test was used, with an a-value of 0.05. RESULTS: Hyper/hypopigmentation and induration/fibrosis were commonly seen at 7 months follow-up (p = 0.84 and 0.83, respectively). The cosmetic results were good or excellent. CONCLUSION: Although the present study included a small number of patients and short follow-up time, the preliminary results were comparable between the study groups. PMID- 19301735 TI - Application of flow cytometric beads for simultaneous CD4 and CD8 determinations in HIV-1 infected thalassemia patients. AB - BACKGROUND: CD4 and CD8 are determined by either a dual step assay using a calculation of absolute lymphocytes obtained from routine CBC or by a single step assay by including cytometric beads and fluorescent microspheres in the sample so that an absolute cell count can be made, simultaneously. Since thalassemia is common in Thailand, and a number of nucleated red blood cells (NRBC) are observed in peripheral blood of thalassemia patients with severe anemia, we speculated that NRBC in HIV-1 infected thalassemia patient with severe anemia might cause an error in CD4 and CD8 determinations in the dual step assay. OBJECTIVE: Comparing cytometric beads in three-color-lyse-no-wash in the single step assay in CD4 and CD8 with dual step assay by calculation using absolute lymphocytes obtained from routine CBC in HIV-1 infected thalassemia patients with severe anemia. MATERIAL AND METHOD: Simultaneous screening of alpha (SEA type)- and beta thalassemia using a multiplex PCR was done. In the thalassemia patients with severe anemia having 13-1,392 NRBC/100 WBC, it found significant differences (p < 0.001-0.002, paired-t-test) of the means of cytometric bead CD4 and CD8 and the means of NRBC corrected CD4 and CD8 as compared to the means of NRBC uncorrected CD4 and CD8 in dual step determinations. In the thalassemia patients with lesser severe anemia, having less than 10 NRBC/100 WBC, there were no significant differences (p > 0.05, paired-t-test) of the means of cytometric bead CD4 and CD8 and the means of NRBC corrected CD4+ and CD8+ as compared to the means of NRBC uncorrected CD4 and CD8 in dual step assay. In comparison of CD4 and CD8 determinations in HIV-1 infected thalassemia patients with severe anemia having more than 10 NRBC/100 WBC, there were significant differences (p < 0.002, paired-t-test) of the means of cytometric bead CD4 and CD8 and the means of NRBC corrected CD4 and CD8 as compared to the means of NRBC uncorrected CD4 and CD8 in dual step assay. CONCLUSION: Results indicated that the NRBC in HIV-1 infected or uninfected thalassemia with severe anemia having more than 10 NRBC/100 WBC do cause an error in CD4 and CD8 determinations in dual step in routine assay. Therefore, either cytometric beads application in the single step or the conventional calculation using NRBC corrected absolute lymphocytes in three-color-lyse-no-wash assay is essentially needed in the flow cytometric assayfor CD4 and CD8 determinations. PMID- 19301736 TI - Paraoxonasel phenotype distribution in Thais. AB - OBJECTIVE: To determine the PON1 activity and phenotype distribution in a Thai population. STUDY DESIGN: Prospective Descriptive study. MATERIAL AND METHOD: Between October 2001 and April 2002, 160 healthy Thai individuals aged 20-74 years were assessed for PON1 activity and the phenotype distribution by using dual substrate method. RESULTS: The means +/- SD of basal, salt-stimulated paraoxonase and arylesterase activities were 239.7 +/- 83.9 nmol/min/mL 555.2 +/- 222.2 nmol/min/mL and 147.6 +/- 33.8 micromol/min/mL respectively. The authors observed a wide interindividual variability up to 6.9-fold for paraoxonase activity and 4.6-fold for arylesterase activity. The authors found a range of ssPON/ARE ratio from 1.04 to 7.05 and three distinctive phenotype modals of AA (1.04-2.25), AB (2.44-4.29), and BB (4.53-7.05) with frequencies of 14.4% (AA), 51.9% (AB), and 33.7% (BB). The authors also observed the association of sex with lipid parameters and PON1 activity. CONCLUSION: The distribution of PON1 phenotype in Thais is clearly trimodal with high frequency in BB phenotype. PMID- 19301737 TI - Attributable mortality of imipenem-resistant nosocomial Acinetobacter baumannii bloodstream infection. AB - BACKGROUND: Uncertainty remains concerning the mortality attributable to infections caused by imipenem-resistant acinetobacter baumannii (IRAB). The authors have sought to examine the impact of this resistance on patient mortality. OBJECTIVE: To evaluate the effects of imipenem resistance on the mortality of patients with Acinetobacter baumannii bloodstream infection. MATERIAL AND METHOD: A cohort study was conducted to compare the survival rates between patients with IRAB and imipenem-susceptible A. baumannii (ISAB) bacteremia. RESULTS: The present study shows 35 patients (52.2%) in an IRAB group died in hospital compared to 26 patients (19.9%) in an ISAB group (p < 0.001). Multivariate analysis using Cox's proportional hazard model for controlling the confounding effects due to the severity of underlying diseases, inappropriate antibiotic treatment, and primary source of bacteremia show no statistically significant difference in mortality rates between the two groups. CONCLUSION: The observed higher mortality rate among patients with an IRAB bloodstream infection may not be attributable to imipenem resistance but may in some part be due to a more severe illness, inappropriate antimicrobial therapy, and primary source of infection. PMID- 19301738 TI - Posttraumatic stress disorder of the Tsunami survivors in Thailand. AB - BACKGROUND: The natural disaster known as "the Tsunami" occurred in the Andaman sea coast of Thailand in December 2004, and there had been questions whether it could cause PTSD amongst the population who lives in the affected area and how to avoid PTSD condition to occur. OBJECTIVE: Establish statistical results of psychosocial factors, and their correlation to PTSD and other mental disorders to generate the PTSD database. MATERIAL AND METHOD: A cross-sectional community surveys from 3,133 samples had been conducted in two phases from the same sampling group. The first phase was concerned with prevalence of PTSD, depression, and related factors. The second phase included 2,573 samples from the first phase and focused on chronic PTSD and other mental disorders. RESULTS: The 3,133 samples used in the first phase show that 33.6% suffered from PTSD, 14.27% with depression, and 11.27% suffered from both. The 2,573 samples from the first phase were followed, collected the blood, and interview data only 21.6% were diagnosed with chronic PTSD. CONCLUSION: The statistical analysis has identified risks factors that could cause PTSD, and protective actions that could help to prevent PTSD. The prevalence of PTSD was still higher in the affected region six months after the Tsunami. PMID- 19301739 TI - Influence of child rearing by grandparent on the development of children aged six to twelve years. AB - OBJECTIVE: To investigate the influence of child rearing by grandparent on the development of children aged six to twelve years. MATERIAL AND METHOD: A cross sectional study was conducted in 320 children that were cared for by a parent and grandparent selected by cluster sampling. The data were collected between March 10 and April 8, 2006 by questionnaire about child and family factors. The TONI III test was used to test the child development. Data were analyzed by frequency distribution, logistic regression, and multiple logistic regression. RESULTS: Child caregiver had a significant influence on child development (p-value < 0.05). Children reared by a grandparent had 2.0 times higher chance of having delayed development compared with those who were reared by the parent. In addition, significant family factors that had impact on the child development were child rearing and family income. CONCLUSION: Child rearing by a grandparent had 2.0 times higher chance of having delayed development than those reared by the parent. Therefore, family and health personnel should plan to ensure the development and learning process of children that are cared by the grandparent. PMID- 19301740 TI - Information use behavior of clinicians in evidence-based medicine process in Thailand. AB - OBJECTIVE: To investigate the information-use behavior of Thai clinicians in the evidence-based medicine (EBM) process. MATERIAL AND METHOD: Based on the survey research, 198 questionnaires were sent to EBM clinicians working in public hospitals in Thailand. The data were analyzed by mean, percentage, and factor analysis. RESULTS: One hundred and fifty-seven questionnaires (79.3%) were returned. The results revealed that 52.9% of the clinicians used EBM process in clinical practice at a high level and 41.4% at a moderate level. Most respondents (91.7%) used information for supporting their teaching and learning process as well as for professional self-development. About two-third used information for supporting their clinical decision. The types of information that the clinicians used in high percentage were research articles from medical journals (89.7%), systematic reviews (83.4%), textbooks, and reference books in the medical field (80.8%). The information resources that were often used including Internet resources (84.1%), hospital or medical school libraries (73.7%), expert consultation (59.8%), and the medical record unit (41.9%). Most of the respondents (89.7%) used PubMed, search engine (85.6%) and Cochrane Library (56.4%) as the tools for accessing information. Most respondents frequently had accessed to information 2-3 days a week and 93.7% of them preferred to access information resources via the Internet by themselves at their office or home. For searching strategies, most clinicians used key words (95%). In the present study, 20 variables were designed to test the factors correlated with the clinicians' information use. The results showed that the six variables (information use, EBM use, experience, organization, competency, and educational background) were significantly correlated with information used by clinicians in EBM process. CONCLUSION: Most Thai clinicians in the present study used EBM process. They regularly searched information by themselves with simple strategy. The results of the present study could be used for planning to improve the quality of clinicians in EBM practice. Since information use is important in using EBM, all hospitals should have adequate facilities to provide medical information for clinical practice. Relevant data from the present study may be useful for planning the use of EBM process and to further researches. PMID- 19301741 TI - Plant pathology. Techniques and protocols. Preface. PMID- 19301742 TI - Bioassay for diagnosis of plant viruses. AB - Bioassay provides a useful means of detecting and identifying plant viruses. The choice of procedure and environmental conditions for the test depends on the virus under investigation. PMID- 19301743 TI - Development of enzyme linked, tissue blot and dot blot immunoassays for plant virus detection. AB - Immunoassays are among the most powerful and useful techniques for analysis of biological materials. There are numerous variations in which immunoassays can be performed. Coupled with enzyme, using chromogenic substrates, the enzyme immunoassay technique is used to trace the target antigen in tissues. The technique also is used to measure the concentration of antigen in tissue extracts. This chapter provides fundamental information that is needed to carry out the routinely used procedures in plant virus research. PMID- 19301744 TI - Immunisation strategies for antibody production. AB - A range of immunisation techniques can be used for the successful production of antibodies. The choice of method used is dependent on the nature of the antigen and the type of antibody required by the user. PMID- 19301745 TI - Preparation of immunogens and production of antibodies. AB - The quality of reagents greatly affects the interpretation of serological tests. Methods used in conventional viral purification and molecular cloning and expression of target viral proteins to obtain antigens for immunization are presented. Immunization of rabbits, mice and chickens and isolation of immunoglobulin from immunized animals also are described. PMID- 19301746 TI - Enzyme-linked immunosorbent assay detection of Ralstonia solanacearum in potatoes: the South African experience. AB - Outbreaks of bacterial wilt caused by Ralstonia solanacearum in South Africa in the 1980s necessitated the development of a sensitive assay for its detection. In this chapter, the development of an enzyme-linked immunosorbent assay (ELISA) for the detection of Ralstonia solanacearum including antibody production, the ELISA method itself, potato sample preparation for testing, ELISA result validation and interpretation, and confirmation of infection are discussed. Data showing the drop in the Ralstonia solanacearum infection rate, after testing was implemented, are presented and shows how the use of this method has brought the disease under control in South Africa. PMID- 19301747 TI - Production of monoclonal antibodies to plant pathogens. AB - The use of monoclonal antibodies in plant pathology has improved the quality and specificity of detection methods for diseases. Hybridoma technology allows the limitless production of highly specific antibodies which can be used to identify pathogens to the species or even sub-species level. PMID- 19301748 TI - Enzyme-linked immunosorbent assay for the detection and identification of plant pathogenic bacteria (in particular for Erwinia amylovora and Clavibacter michiganensis subsp. sepedonicus). AB - Enzyme-linked immunosorbent assay (ELISA) is the most commonly used serological diagnostic technique. A number of different ELISA formats can be used for the detection of bacterial plant pathogens and in particular Erwinia amylovora and Clavibacter michiganensis subsp. sepedonicus. PMID- 19301749 TI - Indirect immunofluorescence microscopy for the detection and identification of plant pathogenic bacteria (in particular for Ralstonla solanacearum). AB - Immunofluorescence microscopy is a very sensitive serological test which harnesses both the power of antibodies to bind to targets along with the use of the fluorescence microscope to visualise the structures to which they bind. Antibody binding is visualised by the fluorescent emission from a marker molecule bound to the antibody. The technique is of particular use in bacteriology and pathology where the location and morphology of the bacterial cells can be viewed along with the location of the fluorescently labelled antibodies. PMID- 19301750 TI - Detection of four major bacterial potato pathogens. AB - Ring rot, brown rot and blackleg represent major bacterial pathogens of potato. The methods described below are aimed at basic identification of ring rot, brown rot or blackleg in a tuber sample. PMID- 19301751 TI - Erwinia amylovora: modern methods for detection and differentiation. AB - Erwinia amylovora is the causative agent of fire blight, a very destructive disease of numerous members of the rosaceae. The primary route of infection for host species, including commercially grown apple and pear, is the newly opened blossom. Susceptibility of flowers to infection for only a few days creates narrow window for infection. Not surprisingly, the risk of disease is related to E. amylovora population size. As a result, methods that supply quick, accurate and sensitive quantification of the pathogen population are important tools for determining the need for and the efficacy of disease control intervention. Plating samples and assessing colony-forming units constitutes an accurate and sensitive but slow method. Endpoint PCR is quick and sensitive but is not particularly amenable to quantification. We describe a real-time PCR procedure that provides all requirements. This method is based on chromosomal genes rather than on the pEa29 plasmid and so can be used to measure isolates that have been cured of the plasmid. The method has been used very successfully in directly quantify whole E. amylovora cells, in a variety of tissues from the orchard environment. PMID- 19301752 TI - The use of fluorescent in situ hybridization in plant fungal identification and genotyping. AB - FISH is a widely used technique in many laboratories not only for cytogenetic studies, but also in other biological fields. It requires a combination of skills in molecular biology, cytogenetics, immunocytochemistry, microscopy and cellular imaging analysis. PMID- 19301753 TI - Use of molecular methods for the detection of fungal spores. AB - Traditional methods for the isolation and identification of fungal spores can be time-consuming and laborious. DNA-based methods for fungal detection can be used to detect the spores of plant-pathogenic fungi. Air borne spores can be collected and identified by PCR allowing identification of the species. PMID- 19301754 TI - Identification of Phytophthora fragariae var. rubi by PCR. AB - The following chapter describes a PCR method for the identification of the raspberry root rot pathogen Phytophthora fragariae var. rubi. Furthermore, a nested PCR suitable for the detection of the pathogen in infected raspberry roots and validated against the "Duncan bait test" (EPPO Bull 35:87-91, 2005) is explained. Protocols for different DNA extraction methods are given which can be transferred to other fungal pathogens. PMID- 19301755 TI - Detection of double-stranded RNA elements in the plant pathogenic fungus Rhizoctonia solani. AB - Many species of fungi have been shown to harbor double-stranded RNA (dsRNA) elements. A single fungal isolate of Rhizoctonia solani may have as many as five different dsRNA elements within them. The presence of specific dsRNA elements influence pathogenicity in host plants. PMID- 19301756 TI - Immunocapture-PCR for plant virus detection. AB - Immunocapture followed by the detection of viruses using polymerase chain reaction is a versatile, sensitive and robust diagnostic technique. The application of this hybrid method of virus detection in plants is particularly useful in species or tissues containing inhibitory substances. In addition, antibody-mediated virus purification is usually simpler than other methods of isolation. PMID- 19301757 TI - Multiplex polymerase chain reaction (PCR) and real-time multiplex PCR for the simultaneous detection of plant viruses. AB - Multiplex Polymerase Chain Reaction (PCR) can be used for the simultaneous detection of plant viruses. Multiple primer pairs or polyvalent primer pairs can be used to detect and identify several viruses in a single PCR. PMID- 19301758 TI - Fluorescent-based techniques for viral detection, quantification, and characterization. AB - Fluorescent-based technologies offer opportunities for developing new assays for detection, quantification, and characterization of viral isolates. According to the intrinsic characteristics of fluorescent-based tools (high specificity, sensitivity, and reliability), such type of molecular assays makes possible investigations on original studies such as evolutionary processes (including fitness measurement of isolates), quantitative epidemiology, or the analysis of synergism and antagonism between closely related isolates. The development of these tools is very simple and requires, in complement to basic molecular knowledge such as extraction, cloning, and (RT)-PCR procedures, only the identification of short specific sequence(s) in the targeted viral genome. The Single Nucleotide Polymorphism (SNP) and the 'real-time' RT-PCR assays are proposed as fluorescent-based tools for qualitative and quantitative viral detection, respectively. Moreover, the SNaPshot technology is described as method for isolate characterization. PMID- 19301759 TI - Analysis of population structures of viral isolates using single-strand conformation polymorphism method. AB - The analysis of viral populations requires the use of techniques that describe characteristics of individuals. The single-strand conformation polymorphism (SSCP) makes possible the identification of genetic differences between viral sequences and constitutes an alternative to the expensive and time-consuming cloning and sequencing strategies. Applied to small genomic regions (from 100 to 500 bases in length), SSCP patterns could describe, under appropriate experimental conditions, single nucleotide variations in the studied sequence. The different steps of a complete SSCP procedure, from sampling to pattern analysis (including nucleic acid extraction, RT-PCR amplification, double stranded DNA quantification, polyacrylamide gel preparation, electrophoresis conditions, and staining procedures), are described using a region (500 bases) of the barley yellow dwarfvirus-PAV (BYDV-PAV, Luteovirus) genome as molecular target. PMID- 19301760 TI - Direct detection of plant viruses in potato tubers using real-time PCR. AB - Virus indexing of seed potatoes can be carried out by growing eye plugs to produce small plants and then testing them by ELISA, but this method is time consuming. Direct testing of the eye plugs by ELISA is not reliable, and so a method has been developed for the routine testing of seed potatoes for virus by PCR. PMID- 19301761 TI - An accelerated soil bait assay for the detection of potato mop top virus in agricultural soil. AB - An accelerated soil bait test can be used to determine whether a field harbours virus-carrying Spongospora subterranea. S. subterranea is the causal agent of powdery scab and also the only vector of potato mop top virus (PMTV). Real-time RT-PCR can detect PMTV RNA in the roots of bait plants after 2 weeks of growth in viruliferous soil. This test may be used to assess the risk of planting potato crops in a particular field. PMID- 19301762 TI - Detection of phytoplasmas of temperate fruit trees. AB - Phytoplasmas are associated with hundreds of plant diseases globally. Many fruit tree phytoplasmas are transmitted by insect vectors or grafting, are considered quarantine organisms and a major economic threat to orchards. Diagnosis can be difficult, but immunochemical and molecular methods have been developed. PMID- 19301763 TI - PCR detection of potato cyst nematode. AB - Potato cyst nematode (PCN) is responsible for losses in potato production totalling millions of euros every year in the EC. It is important for growers to know which species is present in their land as this determines its subsequent use. The two species Globodera pallida and Globodera rostochiensis can be differentiated using an allele-specific PCR. PMID- 19301764 TI - Potato cultivar genome analysis. AB - Due to the yearly increase in the numbers of new potato varieties obtaining Plant Breeders' Rights the reliable maintenance of large culture collections of reference varieties for DUS testing is becoming more and more difficult, as accidental mix ups might occur. Efficient identification methods and databases can act as an aid to overcome this problem. Identification of cultivars by morphological characteristics is a highly skilled and time-consuming task, and for these reasons a rapid and robust method for variety differentiation has become extremely desirable. By use of a set of nine microsatellite (SSR) markers we can differentiate over 1,000 cultivars, including the majority of varieties on the European Union Common Catalogue, but excluding somaclonal variants (e.g. Red King Edward and King Edward) and mutants. The whole identification process from DNA extraction to accurate identification can be carried out in a single day. PMID- 19301765 TI - Barley variety identification using SSRs. AB - There is a current and developing need for rapid and accurate methods of barley varietal identification which go beyond traditional morphological analysis. Methods using DNA analysis have the capacity to fulfil this role with microsatellites being the current marker of choice. The majority of barley cultivars on the National List can be differentiated using 6 SSRs and bulk samples, using the methods described here. PMID- 19301766 TI - [Papillomavirus and cervical cancer in Chile]. AB - Molecular, clinical and epidemiological studies have established beyond doubt that human papiloma viruses (HPV) cause cervical cancer. The virus is also associated with genital warts and other less common cancers in oropharynx, vulva, vagina and penis. Worldwide, VPH genotypes 16 and 18 are the most common high risk genotypes, detected in near 70% of women with cervical cancer. The discovery of a cause-effect relationship between several carcinogenic microorganisms and cancer open avenues for new diagnostic, treatment and prevention strategies. In this issue of Revista Medica de Chile, two papers on HPV are presented. Guzman and colleagues demonstrate that HPV can be detected in 66% to 77% of healthy male adolescents bypolymerase chain reaction and that positivity depends on the site of the penis that is sampled. These results support the role of male to female transmission of high risk HPVs in Chile and should lead to even more active educational campaigns. The second paper provides recommendations for HPV vaccine use in Chile, generated by the Immunization Advisory Committee of the Chilean Infectious Disease Society. To issue these recommendations, the Committee analyzes the epidemiological information available on HPV infection and cervical cancer in Chile, vaccine safety and effectiveness data, and describes cost effectiveness studies. Taking into account that universal vaccination is controversial, the Committee favors vaccine use in Chile and it's incorporation into a national program. However, there is an indication that the country requires the implementation of an integrated surveillance approach including cross matching of data obtained from HPV genotype surveillance, monitoring of vaccination coverage, and surveillance of cervical cancer. The final decision of universal vaccine use in Chile should be based on a through analysis of information.ev Mid Chile PMID- 19301767 TI - [Interactions between beta1 and beta2 adrenergic receptor polymorphisms as risk factors for chronic heart failure]. AB - BACKGROUND: Beta adrenergic receptors (AR) are highly polymorphic and important regulators of cardiovascular homeostasis. Among these, beta1 and beta2 AR regulate cardiac contractility and frequency and are important pharmacological targets. AIM: To evaluate genotype and gene-gene interaction between beta1-AR Arg389Gly and beta2-AR Arg16Gly, Gln27Glu and Thr164Ile polymorphisms, as risk factors for HF. MATERIAL AND METHODS: Eighty chronic HF patients and eighty-eight controls matched by age and sex were genotyped for beta1-AR Arg389Gly, beta2-AR Arg16Gly, Gln27Glu and Thr164Ile polymorphisms. RESULTS: The presence of beta2-AR Glu allele was a risk predictor for HF (odds ratio (OR) = 2.81; 95% confidence intervals (CI) = 1.49-5.31). Interactions that increased the risk for HF were found in patients carrying at least one of the beta2-AR Glu and beta2-AR Gly allele (OR = 3.81; 95% CI = 1.50-0.70) and beta2-AR Glu and beta1-AR Gly allele combination (OR = 5.51; 95% CI = 2.19-13.86). Furthermore, the frequency of beta2 AR Glu allele was higher among patients with a history of acute myocardial infarction (with infarction: 0.534, without: 0.313, p = 0.01). CONCLUSIONS: Beta2 AR Glu allele could be a risk predictorfor HF. This risk could be enhanced by the additional presence of beta2-AR Gly16 or beta1-AR Arg389 alleles. The frequency of beta2-AR Gln27 Glu allele was higher among patients with a history of myocardial infarction. PMID- 19301768 TI - [Prevalence of human papillomavirus genital infection among male university students]. AB - BACKGROUND: Human papillomavirus (HPV) infection is the most common sexually transmitted disease. AIM: To determine prevalence of HPV genital infection in voluntary asymptomatic male university students. MATERIAL AND METHODS: A cross sectional study in 62 asymptomatic, sexually active male students. Exfoliated cells were obtained from the penile shaft and coronal sulcus. Samples were analyzed for HPV DNA detection and genotyping by polymerase chain reaction and Reverse Line Blot. RESULTS: The prevalence of HPV infection was 84%. HPV detection was 77% in penile shaft and 66% in coronal sulcus. The most commonly detected types were HPV-16 (45%), HPV-11 (19%), HPV-6 (10%) and HPV-18 (9%). Multiple infection was found in 54%. The most frequent combinations were VPH11/16 (18%) and VPH16/18 (5%). CONCLUSIONS: HPV infection is highly frequent in asymptomatic male university students, high risk HPV types were greatly predominant. PMID- 19301769 TI - [Melatonin reduces cortisol response to ACTH in humans]. AB - BACKGROUND: Melatonin receptors are widely distributed in human tissues but they have not been reported in human adrenal gland. AIM: To assess if the human adrenal gland expresses melatonin receptors and if melatonin affects cortisol response to ACTH in dexamethasone suppressed volunteers. MATERIAL AND METHODS: Adrenal glands were obtained from 4 patients undergoing unilateral nephrectomy adrenalectomy for renal cancer. Expression of mRNA MT1 and MT2 melatonin receptors was measured by Reverse TranscriPtase Polymerase Chain Reaction (RT PCR). The effect of melatonin on the response to intravenous (i.v.) ACTH was tested (randomized cross-over, double-blind, placebo-controlled trial) in eight young healthy males pretreated with dexamethasone (1 mg) at 23:00 h. On the next day, at 08:00 h, an i.v. line was inserted, at 08:30 h, and after a blood sample, subjects ingested 6 mg melatonin or placebo. At 09:00 h, 1-24 ACTH (Cortrosyn, 1 microg/1.73 m2 body surface area) was injected, drawing samples at 0, 15, 30, 45 and 60 minutes after. Melatonin, cortisol, cortisone, progesterone, aldosterone, DHEA-S, testosterone and prolactin were measured by immunoassay. RESULTS: The four adrenal glands expressed only MT1 receptor mRNA. Melatonin ingestion reduced the cortisol response to ACTH from 14.6 +/- 1.45 microg/dl at 60 min in the placebo group to 10.8 +/- 1.2 microg/dl in the melatonin group (p < 0.01 mixed model test). It did not affect other steroid hormone levels and abolished the morning physiological decline of prolactin. CONCLUSIONS: The expression of MT1 melatonin receptor in the human adrenal, and the melatonin reduction of ACTH stimulated cortisol production suggest a direct melatonin action on the adrenal gland. PMID- 19301770 TI - [Limited diagnostic testing can decrease the direct economic impact of irritable bowel syndrome]. AB - BACKGROUND: The economic impact of irritable bowel syndrome (IBS) in Mexico in terms of excessive diagnostic testing can be considerably reduced if the recommendations of the Latin American Consensus (LATAM) for IBS are followed. AIM: To estimate the economic impact of IBS in terms of excessive diagnostic testing. MATERIAL AND METHODS: Based on a previously published study, the costs of diagnostic testing for IBS were compared to the theoretical costs according to the recommendations of the consensus. These costs were compared to estimate the economic impact of excessive diagnostic testing. A cost-minimization analysis was also done. RESULTS: For the lowest socioeconomic level in academic medicine, the excessive diagnostic testing had an approximate cost of U.S. $21.38, compared to U.S. $1.72 if the LATAM Consensus recommendations would have been followed, representing a saving of 92.0%. The cost for the highest socioeconomic level in academic medicine was U.S. $1080.36 versus U.S. $103.60 (a saving of 90.4%) and for private medicine, the costs were U.S. $3121.60 versus U.S. $159.90 (a saving of 94.9%) if the recommendations would have been followed. CONCLUSIONS: Limited diagnostic testing recommended by the LATAM Consensus for IBS can significantly decrease the economic impact of this disease in Mexico PMID- 19301771 TI - [Nutritional status and lifestyles of workers from two regions in Chile]. AB - BACKGROUND: Lifestyles and a poor nutritional status are two factors influencing the high prevalence of non communicable chronic diseases among Chilean workers. AIM: To characterize lifestyles and nutritional status of workers from the public and private sector, residing in the Fifth and in the Metopolitan regions of Chile. MATERIAL AND METHODS: Nutritional status, blood pressure, cholesterol, glyeemia, previous medical history, feeding habits and smoking status were assessed in 1036 women and 709 men aged 38 +/- 11 years. RESULTS: Feeding habits were characterized by a low consumption of fruits, vegetables, fish and dairy products and a high consumption of fat and sugar. Forty five percent smoked and 88% were sedentary. Forty two percent were overweight, 18% were obese, 39% had high blood cholesterol, 23% hypertension and 4% diabetes. There was a positive relationship between obesity and hypertension with an odds ratio (OR) adjusted for sex and educational levels of 2.6 (95% confidence intervals (CT) 1.9-3.7) and diabetes with an OR of 2.8 (95% CI 1.6-4.7). Eighty two percent of obese and 28% of overweight subjects underestimated their nutritional status. CONCLUSIONS: A high prevalence of overweight and inadequate lifestyles was found in this population. There was a low self perception of obesity and lack of knowledge about risk factors for chronic diseases. PMID- 19301772 TI - [Nutritional status, body composition and bone mineral density in gastric bypass females: impact of socioeconomic level]. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGBP) has had a posilive impact on co mobidities associated with obesity. However, in the long-term it can induce micronutrient deficiencies. AIM: To petform a complete nutritional assessment in a group of women previously operated of RYGBP from different socioeconomic levels (SEL). PATIENTS AND METHODS: Thirtyy three women (19 high SEL and 14 low SEL), were assessed by dietary recalls, anthropometric measurements, muscle strength, bone mineral density, routine clinical laboratory, semm leeds of vitamin B12, 25OH-vitamin D, folate, calcium, ferritine ceruloplasmin and indicators of bone tutnoter (paratbohormone, osteocalcin and urinary pyridinolines). Their values were compared to those of 30 control women (18 high SEL and 12 low SEL). RESULTS: Low SEL operated women consumed fewer vitamin and mineral supplements compared with their high SEL pairs. No cases of vitamin B12, folic acid or copper deficiencies were detected. Frequency of iron deficiency was similar in patients and controls. Vitamin D insufficiency was higher among patients than in controls (p = 0.047), regardless SEL. Patients had also a higher frequency of high senum P771 and osteocakin and urinary pyridinoline levels. However, no differences in bone mineral density were obseived between operated women and controls. CONCLUSIONS: Vitamin and mineral deficiencies were lower than expected among operated women. However problems associated with vitamin D deficiency were highly prevalent among patients operated of RYGBP, irrespective SEL. These alterations were only detectable through specific markers at this stage, because they did not translate into lower bone mineral density (BMD) of sutgical patients, probably due to the higher pre-operative BMD of these moibid obese patients PMID- 19301773 TI - [Long term survival of patients operated for early gastric cancer]. AB - BACKGROUND: Early gastric cancer involves mucosa and submucosa, independent of lymph node involvement. Radical gastrectomy is the standard treatment. AIM: To assess long term survival of patients operated for an early gastric cancer. MATERIAL AND METHODS: Retrospective review of medical and pathology records of patients subjected to a gastrectomy for an early gastric cancer, between 1975 and 2002. All were treated using a standardized protocol and staged according to 2002 TNM classification of the American Joint Committee of Cancer (AJCC). Demographic and pathologic features, operation performed and long term survival were recorded. Survival was analyzed using Kaplan-Meier method. RESULTS: The series is comprised by 64 males and 41 females aged 61 +/- 1 years. Tumor was located in the upper third of the stomach on 33 subjects and a total gastrectomy was performed in 53. Pathology showed an intestinal type adenocarcinoma in 82 and a diffuse type in 23. In patients with involvement of mucosa and submucosa, 24 +/- 14 and 22 +/- 14 lymph nodes were excised, respectively. Lymph node involvement was present in 8% and 22% of patients with involvement of mucosa and submucosa, respectively. Five years survival was 94% and 78% in patients without and with lymph node involvement, respectively. Survival among patients in stage IA and IB was 94% and 76%, respectively. Multivariate analysis showed that the lymph node involvement was an independent mortality risk factor. CONCLUSIONS: Total gastrectomy in patients with early gastric cancer is associated with a good survival. Lymph node involvement is a mortality risk factor. PMID- 19301774 TI - [Endovascular treatment of type B aortic dissection]. AB - BACKGROUND: Dissections that involve the ascending aorta are classified as type A, regardless of the site of the primary intimal tear, and all other dissections as type B. Type B dissections can have fatal ischemic and hemorrhagic complications. In the chronic state, dilatation and rupture can be mortal. Endovascular surgery is a therapeutic alternative, considering the high rate of complications of conventional surgery. AIM: To report the results of endovascular treatment of type B aortic dissection. MATERIAL AND METHODS: Report of 36 treated patients (30 males) aged 43 to 87 years, with a type B aortic dissection. Seventy eight percent were hypertensive and 39% smoked. The diagnosis was confirmed by CAT scan. Acute patients were treated for complications and chronic patients, for dilatation. In the operating room, an endoprothesis was placed through the femoral artery, to cover the tear. The tear was located and the lumens were differentiated using angiography and transesophageal echocardiography. RESULTS: All procedures were successful. In 16 acute dissections the indications were malperfusion syndrome or unmanageable hypertension in seven patients and imminent rupture or persistent pain in nine. Twenty chronic patients were operated due to dilatation (mean 6 cm). One patient died due to cardiac failure. One patient had a transient paraparesia and two had pulmonary embolism. No patient died in a follow up period ranging from 2.5 to 74 months. Four patients required a new aortic endovascular procedure due to progressive dilatation or endoleak. CONCLUSION: Endovascular treatment of type B aortic dissection has good immediate and long term results. PMID- 19301775 TI - [Validation of a Spanish version of the Decisional Conflict scale]. AB - BACKGROUND: In Chile, in approximately 50% of nursing students, nursing was not their first choice as career. Usually, during the first year, these students must decide whether they would like to continue in the same career. A valid tool is needed to identify decisional conflicts and their contributing factors among these students and to develop an appropriate strategy to support them during their decision-making process. AIM: To translate into Spanish and validate the Generic Decisional Conflict Scale (DCS). MATERIAL AND METHODS: The DCS was translated from English to Spanish and was used with 331 first-year nursing students at the Pontificia Universidad Catolica de Chile. The scale was assessed for validity and reliability using statistical tests, including factor analysis and Cronbach alpha test. RESULTS: The Spanish version of the DCS had acceptable validity and reliability. Factorial analysis identified four factors and only the item: "advice" loaded the other factors. Cronbach alpha was 0.80. CONCLUSIONS: DCS is a valid and useful instrument to identify decisional conflicts and contributing factors to continue studies among nursing students. PMID- 19301776 TI - [Actinomadura madurae mycetoma of the foot. Report of one case]. AB - Mycetoma is a chronic infection that affects skin, subcutaneous tissue and bone. Its etiology can be mycotic or bacterial. It affects mainly the lower extremities of middle age men living in tropical climates. We report a 44-year-old male living in a template zone, consulting for swelling and pain in the left foot, lasting for 10 years. Physical examination showed a swollen left foot with hyperpigmented skin and a few crusted papules. Radiology showed an extensive bone involvement of the midfoot with several oval and radiolucid images. Magnetic resonance showed soft and bone tissue involvement, with multiple oval and low intensity images in T1 and T2. The biopsy was compatible with an unspecific chronic osteomyelitis. A bacterial identiFcation by polymerase chain reaction and sequencing in the biopsy determined the presence of an Actinomadumra madurae. Treatment with cotrimaxazol was started. PMID- 19301777 TI - [Pleural and ascitic fluid infection by Listeria monocytogenes. Report of one case]. AB - Ascitic and pleural fluids infection by Listeria monocytogenes is uncommon. The association of spontaneous bacterial peritonitis and empyema caused by this microorganism has been seldom reported. A 61-year-old male with an alcoholic cirrhosis and an upper right lobectomy for a lung cancer, consulted because of an exacerbation of dyspnea, abdominal pain and fever. Listeria-monocytogenes was isolated from ascitic and pleural fluids and from blood cultures. He was successfully treated with ampicillin and a chest tube for drainage. PMID- 19301778 TI - [Gastric bezoar as complication of gastric banding. Report of one case]. AB - Laparoscopic adjustable gastric banding (LAGB) is used for the management of morbid obesity. Phytobezoars are rarely reported as a complication of this operation and are usually extracted by endoscopic means. We report a 48-year-old male subjected to a gastric banding, that consulted for progressive dysphagia, six months after the operation. A barium meal x-ray examination demonstrated the presence of a bezoar that was dissolved in one week using papain. A control barium meal confirmed the disappearance of the bezoar. PMID- 19301779 TI - [Nutritional genomics: an approach to the genome-environment interaction]. AB - Nutritional genomics forms part of the genomic sciences and addresses the interaction between genes and the human diet, its influence on metabolism and subsequent susceptibility to develop common diseases. It encompasses both nutrigenomics, which explores the effects of nutrients on the genome, proteome and metabolome; and nutrigenetics, that explores the effects of genetic variations on the diet/disease interaction. A number of mechanisms drive the gene/diet interaction: elements in the diet can act as links for transcription factor receptors and after intermediary concentrations, thereby modifying chromatin and impacting genetic regulation; affect signal pathways, regulating phosphorylation of tyrosine in receptors; decrease signaling through the inositol pathway; and act through epigenetic mechanisms, silencing DNA fragments by methylation of cytosine. The signals generated by polyunsaturated fatty acids are so powerful that they can even bypass insulin mediated lipogenesis, stimulated by carbohydrates. Some fatty acids modify the expression of genes that participate in fatty acid transport by lipoproteins. Nutritional genomics has myriad possible therapeutic and preventive applications: in patients with enzymatic deficiencies; in those with a genetic predisposition to complex diseases such as dyslipidemia, diabetes and cancer; in those that already suffer these diseases; in those with altered mood or memory; during the aging process; in pregnant women; and as a preventive measure in the healthy population. PMID- 19301780 TI - [Early rheumatoid arthritis]. AB - This article reviews some key issues of early rheumatoid arthritis such as the dfficulties to recognize this condition during the first months after onset. Therefore, three reference diagnostic criteria have been proposed for any patient presenting with more than three simultaneously inflammed joints, involvement of metacarpophalangeal or proximal interphalangeal joints and morning stiffness lasting more than 30 minutes. Antibodies to cyclic citrullinated peptides are new markers that can be used for diagnosis. The immediate treatment during the "opportunity window" at the onset of inflammation may avoid the erosive joint damage. The use of synthetic or biological disease modifying medications, specially tumor necrosis factor alpha antagonists, also contribute to this purpose. Primary care physicians should be aware of the early signs of the disease to provide an adequate treatment and referral to specialists. PMID- 19301781 TI - [Vascular damage in chronic kidney disease]. AB - Cardiovascular disease is a frequent complication of renal failure and is the most common cause of death in patients with chronic kidney disease (CKD). Accelerated atherogenesis has been widely documented in CKD and diabetic nephropathy is the leading cause of renal failure worldwide. Furthermore, CKD promotes hypertension and dyslipidemia, which in turn may contribute to the progression of renal failure. All together, hypertension, dyslipidemia and diabetes are considered major risk factors for the development of endothelial dysfunction and progression of atherosclerosis. Elevated inflammatory mediators and activation of the renin-angiotensin system contribute through enhanced production of reactive oxygen species, to atherogenesis in CKD. Vascular calcification is also important. Calcification of arteries occurs in the intima in association with atherosclerosis, where it may contribute to plaque formation, and in the media, where it causes stiffening. Increased serum levels of calcification promoters, such as hyperphosphatemia, and a decrease in circulating and local inhibitors of calcification, favor vascular calcification. On the other hand, transdifferentiation of vascular smooth muscle cells to osteblast-like cells would be the pivotal event in calcification. Bone morphogenetic protein agonists and antagonists are playing a role in this osteogenic differentiation. Accelerated atherosclerosis and media calcification will then lead to increased prevalence of coronary artery disease, heart failure, stroke, and peripheral arterial disease. Prevention and treatment of cardiovascular disease are major considerations in the management of individuals with CKD. PMID- 19301782 TI - [Human papillomavirus vaccine. Statement of the Advisory Committee of Immunizations on behalf of the Chilean Infectious Diseases Society. September 2008]. AB - This article briefly reviews the epidemiology of human papillomavirus (HPV) infection and associated diseases globally and in Chile, and the scientific information of the licensed HPV vaccines: Gardasil and Cervari. Considering the available information, the Advisory Committee on Immunizations of the Chilean Society of Infectious Diseases recommends vaccination of teenage girls, ideally before initiating sexual activity, i.e., approximately at the age of 12 to 13 years and vaccination of women of any age if they have not started sexual activity. If women are vaccinated after initiating sexual activity, they should be informed of the lower efficacy of immunization if HPV infection has occurred. Education on responsible sexuality and sexually transmitted diseases should be maintained as a priority. Vaccination should be highly considered for inclusion in the National Immunization Program. PMID- 19301783 TI - [Brugada syndrome a lethal chameleon]. PMID- 19301784 TI - [Cystocerebral syndrome]. PMID- 19301785 TI - [What is internal medicine?]. PMID- 19301786 TI - Osteochondroma in the lumbar intraspinal canal causing nerve root compression. AB - Osteochondromas, which are benign bone tumors that usually develop on long bones, tubular bones, are rarely found in the spine. If they are located in the spinal canal, they may cause nerve root or spinal cord compression, which is a rare but potentially catastrophic manifestation of osteochondromas. In this article, we report a case of a 38-year-old man who presented with low back pain, paresthesia, and weakness of the right lower extremity aggravating gradually for 5 months. No family history of this disease can be traced. The L4-L5 level computed tomography scan showed an abnormal bony protrusion arising from the right interior wall of L5 right lamina toward the intraspinal canal. The protrusion compressed the L5 nerve root severely. T2-weighted magnetic resonance imaging (MRI) of the same level revealed that the L5 nerve root and spinal dura mater were notably compressed by the intraspinal extradural exostosis attached to the right lamina of L5. Considering differential diagnosis, lumbar facet synovial cysts must be excluded as they can also cause myeloradiculopathy with the similar mechanism. The tumor, approximately 6x7x11 mm, was identified after laminectomy of the L5 laminae. Postoperative histopathologic examination confirmed our hypothesis of benign osteochondroma. Postoperatively, the patient recovered rapidly in neurological function and was free of symptoms. Surgery is essential to this rare case. Computed tomography and MRI are helpful for the preoperatively precise indication of tumor extent and its relationships with the adjacent. PMID- 19301787 TI - Fracture of an ossified tibialis anterior tendon. AB - Fractures of ossified tibialis anterior tendon have been successfully managed by the excision of the ossified tendon and transfer of the extensor hallucis longus tendon. A 64-year-old man sustained an injury during mountain hiking 2 weeks prior to presentation, falling down on his heels with his ankle fully plantarflexed. Two tender and bony hard masses were palpable along the course of the tibialis anterior tendon, one at the anteromedial aspect of the ankle and another at the dorsum of the talonavicular joint. Radiographs of the ankle demonstrated an approximately 2x1-cm ovoid-shaped bony mass at the anterior aspect of the ankle joint and another bony mass of similar size and shape at the dorsal aspect of the talonavicular joint. He underwent operative exploration, and complete rupture of the tendon through a bony mass was observed. The gap between the torn ends of the tendon after excision of the mass was too long to be repaired directly. Extensor hallucis longus tendon was retrieved 1 cm proximal to the metatarsophalangeal joint and was passed through the insertion of the anterior tibial tendon and pulled proximally and sutured to itself with the ankle in neutral. At 1 year after surgery, plain radiographs and ultrasonography showed no recurrence of calcification or ossification in the tendon and good mobility of the tibialis anterior muscle was observed. PMID- 19301788 TI - Interpositional arthroplasty of the calcaneocuboid joint using a regenerative tissue matrix to treat recurrent nonunion. AB - Nonunion after foot arthrodesis is a difficult clinical problem to solve. This article presents a case of a patient who underwent 3 unsuccessful attempts at calcaneocuboid joint arthrodesis. This problem was salvaged with interpositional arthroplasty of the joint with successful clinical outcome. A 49-year-old woman with longstanding rheumatoid arthritis underwent triple hindfoot arthrodesis. Although the talonavicular joint and the talocalcaneal joints achieved successful arthrodesis, the calcaneocuboid joint did not unite. Revision arthrodesis of the joint, as well as another attempt at percutaneous grafting had failed. Infection was ruled out by biopsy. In each instance, her postoperative course was complicated by difficulty complying with nonweight bearing restrictions, changes in her rheumatoid arthritis medications, and medical comorbidities. Considering all of her medical and rehabilitation issues, she underwent interpositional arthroplasty using a regenerative tissue matrix, rather than a fourth attempt at arthrodesis. Allograft dermal matrix was used as interpositional material at the calcaneocuboid joint. One year postoperatively, the patient reports only mild discomfort in the hindfoot, which does not interfere with her activities of daily living. In the difficult setting of multiple failed attempts at arthrodesis, interpositional arthroplasty can be considered in a nonweight-bearing joint such as the calcaneocuboid joint. Interpositional arthroplasty represents a technique capable of providing pain relief in clinical situations in which osseous union cannot be achieved. PMID- 19301789 TI - Uncovered posterolateral rotatory elbow instability with cubitus varus deformity correction. AB - The relationship between a longstanding cubitus varus deformity and the delayed onset of posterolateral rotatory instability has casually been reported as being caused by abnormal repetitive varus and external rotation torques with progressive failure of the lateral collateral ligament. This article presents a case of longstanding cubitus varus in a young man, following a supracondylar fracture sustained during childhood. Posterolateral rotatory instability with lateral collateral ligament deficiency was discovered after a corrective distal humeral realignment osteotomy. The scenario was worsened by medial olecranon overgrowth that predicated symptomatic medial snapping triceps during terminal extension and secondary ulnar neuropathy. Lateral collateral ligament reconstruction as well as an extra-articular olecranon osteotomy was performed. The ligament was reconstructed with a free graft harvested from the central portion of the distal triceps and secured with 2 bioabsorbable screws. The patient returned to work with no residual symptoms at final follow-up. This case highlights the possibility of posterolateral rotatory instability even with no clinical evidence at presentation. It suggests the importance of laterally redirecting the triceps vector and reconstructing the lateral collateral ligament as additional procedures at the time of supracondylar osteotomy. PMID- 19301790 TI - Total hip arthroplasty in a patient with a contralateral hemipelvectomy. AB - Hemipelvectomy is a rare procedure performed for tumor resection or trauma. The limb may be spared, and the patients often require extensive rehabilitation. We report the outcome of a total hip arthroplasty (THA) in a patient with endstage hip arthritis in a high dislocated hip contralateral to a remote, traumatic hemipelvectomy. To our knowledge this is the first reported case with mid-term follow-up of a patient undergoing THA for degenerative, traumatic hip dislocation with a contralateral external (limb-sacrificing) hemipelvectomy. Patient history, surgical details, and follow-up 5 years from the index arthroplasty are presented. This case report documents that THA after hemipelvectomy can be a durable procedure, and provide reliable pain relief and improvement in function at intermediate follow-up using good surgical techniques, and with the use of noncemented implants. PMID- 19301791 TI - Protrusio of a ceramic femoral head through the acetabular metallic shell, extensive metallosis and 'bubble sign'. AB - A 24-year-old patient with a history of juvenile rheumatoid arthritis underwent a primary cementless left total hip arthroplasty (THA). The original THA consisted of an Optifix 54 cup with a 3-mm thick polyethylene liner, an Optifix size 4 stem (Smith & Nephew Richards, Memphis, Tennessee) and a Biolox aluminum 32-mm femoral head. Fourteen years later, radiographs demonstrated extensive wear of the polyethylene liner resulting in direct articulation and abrasion wear of the ceramic femoral head on the cup and a bubble sign. This article presents a case of a catastrophic failure of a ceramic/polyethylene bearing with destruction of the polyethylene liner and the metallic shell and protrusio of the nonfractured ceramic head through the metallic shell. To our knowledge this is the first description of extensive metallosis and subsequent radiograph bubble sign not presenting as a result of wear of a metal-on-metal articulation. At the time of revision surgery-Hydrocel TNT Monoblock 58 cup (Zimmer, Warsaw, Indiana), Wagner 265/14 stem (Zimmer), and a Co/Cr 28-mm head-copious metallic debris was seen both macroscopically and histologically, with the ceramic head protruding behind the metallic shell. Multiple factors may have been responsible for this failure including a thin polyethylene shell, a suboptimal locking mechanism, gamma in air sterilization for polyethylene, multiple screw-holes that reduce the contact surface between shell and polyethylene, the rough surface on the inside of the shell and non-articular wear at the metal polyethylene interface within the acetabular component and the high demands of this active young patient. PMID- 19301792 TI - Calcific tendinitis of popliteus tendon: arthroscopic excision and biopsy. AB - Calcific tendinitis results from the deposition of calcium hydroxyapatite crystals in periarticular muscular attachments. It is a rare cause of knee pain commonly affecting patients aged 40 to 70 years. Although commonly seen in the shoulder, it should be kept in mind in nontraumatic cases, particularly when the pain is severe and localized to the lateral aspect of the knee. The exact mechanism of hydroxyapatite deposition is unclear, although genetic and metabolic factors have been suspected. A 45-year-old man presented with severe pain in the lateral aspect of his knee with local tenderness over the lateral epicondyle. Radiographs revealed multiple calcific deposits just below the lateral epicondyle of the femur. Magnetic resonance imaging showed multiple areas of low-signal present intra-articularly near the popliteus tendon that was suspected to be calcification. Erythrocyte sedimentation rate and C-reactive protein were slightly raised and other blood investigations including uric acid were within normal limits. Due to failure of conservative treatment, arthroscopy was performed through standard anteromedial and anterolateral portals. Arthroscopy revealed reddish synovial congestion in the lateral gutter. Partial synovectomy was performed with a shaver through a superolateral portal and the calcific deposit was found to lie between the popliteus tendon and the lateral collateral ligament. This was excised and sent for biopsy. Histopathological evaluation revealed the presence of hydroxyapatite crystals within degenerated tendon thereby confirming the diagnosis of calcific tendinitis. Immediate resolution of symptoms following excision allowed the patient to perform activities of daily living immediately postoperatively without pain. PMID- 19301793 TI - Incarcerated tibial nail. AB - Removal of intramedullary nails is considered a routine procedure but may prove to be challenging. Bone ongrowth or overgrowth, damage to the proximal threads of the nail, and broken nails or locking screws may complicate intramedullary nail removal. This article presents a case of a 28-year-old patient with an incarcerated tibial nail and describes a salvage procedure for tibia nail extraction after all previously described methods have failed. The authors recommend that no excessive force be used to extract an incarcerated nail but to approach the problem in a step-wise fashion. If the nail is still incarcerated and will not move, the tibia needs to be completely open by removing a one-third of circumference longitudinal bone window. It is important to discuss all the possible options with the patient during the consent. The patient may not be aware of the difficulties that can be encountered during nail removal and the fact that it may necessitate a major procedure followed by a prolonged recovery time. It is important to ascertain the reason for nail removal and whether it is necessary to implement even drastic measures to do so. Infection, nonunion, deformity or refracture requiring fixation are indications for nail removal. PMID- 19301794 TI - Outcome and complications of constrained acetabular components. AB - Constrained acetabular liners were developed for the surgical treatment of recurrent instability by holding the femoral head captive within the socket. This article summarizes the data describing constrained component designs, indications, outcome, and complications. Different designs accept head sizes of varying diameter and have differing amounts of rim elevation and offset, allowing slight variations in the range of movement allowed. Complications of constrained acetabular components can be divided into three categories. The first category is directly related to the constraining mechanism such as dislocation, head dissociation from the stem, liner dissociation from the acetabular device, and impingement with or without locking ring breakage. The second category is related to increased constraint such as aseptic component loosening and osteolysis and periprosthetic fracture. The third category includes those cases not associated with increased constraint such as infection, deep vein thrombosis, and periprosthetic fracture. This device is effective at achieving hip stability, but the complications related to the constraining mechanism and increased constraint are of concern. These devices should be used as a salvage measure for the treatment of severe instability. PMID- 19301795 TI - Radial nerve palsy associated with humeral shaft fractures. AB - Radial nerve palsy frequently accompanies humeral shaft fractures. The need for operative exploration in different situations continues to be debated. PMID- 19301796 TI - Peptic ulcer disease: clinically relevant causes and treatments. AB - Peptic ulcer disease is a significant cause of morbidity and in certain cases mortality among affected individuals. Proper identification and treatment of peptic ulcer disease is imperative to decreasing its associated sequellae. The most common causes of peptic ulcer disease are the use of nonsteroidal anti inflammatory drugs (NSAIDs) and infection with Helicobacter pylori. Initial assessment of the patient with dyspepsia is paramount, as the presence of symptoms will dictate further management. With currently available treatment regimens and the ability to reduce gastrointestinal bleeding it is important for all clinicians to have knowledge of this disease, its diagnosis, and pharmacotherapy. PMID- 19301797 TI - The effect of intra-articular epinephrine lavage on blood loss following total knee arthroplasty. AB - The purpose of this study was to examine the effects of intra-articular epinephrine lavage on blood loss following total knee arthroplasty (TKA). Our retrospective study involved 189 patients who had primary cemented TKA by 1 of 2 surgeons. Surgeon 1 performed 41 procedures without and 53 with the epinephrine lavage. Surgeon 2 performed 41 procedures without and 54 with the epinephrine lavage. We compared calculated blood loss (CBL) for the patients in all 4 groups. We found no significant differences in CBL among the patients in the 4 patient groups. Our results show that an intra-articular epinephrine lavage does not affect blood loss after TKA. PMID- 19301798 TI - Anatomical graft passage in transtibial posterior cruciate ligament reconstruction using bioabsorbable tibial cross pin fixation. AB - There is no consensus on how to optimally reconstruct a posterior cruciate ligament (PCL). This paper introduces a new surgical technique that provides more secure fixation, better graft selection, and more anatomic graft passage. This study focused on the tibial backside fixation of a soft tissue graft using a rigidfix system (RIGIDfix system; Mitek, Johnson & Johnson, Raynham, Massachusetts) and anatomic graft passage anterior to the meniscofemoral ligament (MFL) of Wrisberg. PMID- 19301799 TI - Fate of mu receptors during rat skeletogenesis. AB - Studies have intimated a role for endogenous opioids in skeletal ontogeny. We hypothesized that this role may be confined to the perinatal period. We, therefore, examined both fetal and postnatal rat long bones to determine the pattern, if any, of mu receptor expression. Perfused long bones and brains were harvested from 3 to 4 each of near-term fetuses, 3- to 4-week-old neonates, and 6 week-old juveniles. Tissues were decalcified, embedded, sectioned, quenched of peroxidase, and then blocked. Sections were incubated overnight with either rabbit antirat mu receptor IgG or naive rabbit IgG (control) at 4 degrees C. The next day, sections were washed, blocked again, and incubated with biotin-labeled secondary antibody, streptavidin-peroxidase conjugate, and a chromogen substrate with intervening wash steps. Slides were counterstained with hematoxylin and coverslipped. Digital photomicrographs were then imported into an image analysis program and the percent area of stained cortical and trabecular bone quantified. Mu receptor expression decreased significantly with age. Approximately 25% of the area of fetal long bones stained positively, including the endosteum, periosteum, and the growth plate. Little or no nonspecific staining occurred. Staining in neonatal tissue was diminished to <11% of the area and involved areas of apparent remodeling; chondrocytes in the growth plates failed to stain. Finally, juvenile bone evidenced staining approaching background levels produced by control slides (approximately 2%). Mu receptors are abundant in developing rat long bones during fetal development but become progressively less abundant postnatally. This infers a role for endogenous opioids during skeletal ontogeny. PMID- 19301800 TI - Antifibrinolytic therapy in complex spine surgery: a case-control study comparing aprotinin and tranexamic acid. AB - A case-control study was performed to determine the impact of aprotinin or tranexamic acid use on reducing intraoperative blood loss and transfusion needs in complex spine surgery. Sixty-nine patients undergoing complex spine surgery received aprotinin or tranexamic acid. The aprotinin group contained 30 patients (8 men and 22 women) and the tranexamic acid group 39 patients (11 men and 28 women). The following variables were recorded: duration of surgery, number of levels fused, intraoperative and total blood loss, and number of blood units transfused (autologous and allogenic). In addition, various parameters related to blood loss in this type of surgery were calculated. The groups differed with regard to duration of surgery (aprotinin 662 min vs tranexamic acid 448 min, P<.001) and number of levels fused (aprotinin 11.2 vs tranexamic acid 7.6, P=.004). There were no significant differences in intraoperative blood loss (aprotinin 2118 mL vs tranexamic acid 1608 mL, P=.066) or total blood loss (aprotinin 3312 mL vs tranexamic acid 2627 mL, P=.056). Statistical differences were found for the number of autologous blood units transfused (aprotinin 2.2 vs tranexamic acid 1.3 P=.047) and total units transfused (aprotinin 4.1 vs tranexamic acid 2.6, P=.008). Blood loss per hour of surgery, transfused units per level fused, and transfused units per hour of surgery were similar in the 2 groups. Significant differences were found for intraoperative blood loss per fusion level (aprotinin 228 mL vs tranexamic acid 428, P=.025) and total blood loss per fusion level (aprotinin 360 mL vs tranexamic acid 638 mL, P=.01). Analysis of the applied geometric mean showed that aprotinin reduced total blood loss by 16.4% and total number of blood units transfused by 12.4% as compared to tranexamic acid, although statistical significance was not reached. The type of antifibrinolytic used did not have a significant impact on the main outcome variables of the study. PMID- 19301801 TI - 1150 kyphoplasties over 7 years: indications, techniques, and intraoperative complications. AB - In our cohort of 555 patients with a total of 1150 vertebral fractures treated with kyphoplasty we performed a 30-day postoperative analysis of cement leakage, neurological symptoms, pulmonary embolism, and infections. In our department, 22% of kyphoplasties were performed with calcium phosphate cement and the remainder with polymethylmethacrylate. All patients were initially assessed by an interdisciplinary kyphoplasty colloquium, composed of consultants in traumatology, radiology, and endocrinology. Indications included fresh traumatic vertebral fractures; painful sintered osteoporotic vertebrae; osteolysis and painful vertebral body collapse caused by multiple myelomas; and lymphomas and pathological fractures due to metastases of malignant tumors (prostate cancer, breast cancer, ovarian cancer, and malignant melanoma) or benign vertebral tumors (hemangioma). Contraindications included patients with instability of the posterior wall and/or pedicles, an infection of the fractured vertebra, a severe hemorrhagic diathesis, known allergies to the cements, pregnancy, and ASA score of 4. The standard postoperative computed tomography scan of the kyphoplasty treated vertebrae revealed a dorsal cement leakage in 38 vertebrae representing 3.3% of all levels. A permanent monoparesis of the left leg, 2 cases of temporary neurological deficits, 2 cases of hemorrhage, and 1 asymptomatic pulmonary embolism were observed as postoperative complications. We observed no complications relating to polymethylmethacrylate described in the literature. By careful interdisciplinary indication setting and a standardized treatment model, kyphoplasty presents a safe and effective procedure for the treatment of various vertebral fractures. PMID- 19301802 TI - Percutaneous curettage and bone grafting for humeral simple bone cysts. AB - Traditional treatments for simple bone cysts are associated with significant surgical morbidity and high recurrence rates. The purpose of this study was to evaluate the effectiveness of a percutaneous approach to the treatment of proximal humeral simple bone cysts that includes percutaneous decompression, curettage, and injection of allogeneic cancellous bone combined with autologous bone marrow. Twenty-one children (average age, 10.1+/-4.6 years) with symptomatic simple bone cysts of the proximal humerus were surgically treated with percutaneous cyst decompression, curettage, and cancellous allograft combined with autologous bone marrow. Patients were observed in-clinic with serial examinations and radiographs. Fracture healing and cyst recurrence were determined at follow-up. All patients returned to activities of daily living with full range of motion and were completely asymptomatic. All fractures healed. There were no surgical complications or refractures. Five patients demonstrated radiographic evidence of partial cyst recurrence and required additional surgical treatment. One patient required a third operative procedure for a second recurrence. Three of 8 active cysts demonstrated partial recurrence. Recurrence was associated with younger age. Our goal in the treatment of proximal humeral simple bone cysts is to prevent cyst recurrence and pathologic refracture. Radiographic evidence of cyst resolution occurred in 75% of our patients using percutaneous curettage and bone grafting, and in 95% of patients with a second procedure. As expected, partial recurrence of active cysts after treatment with this method is high. This percutaneous technique is a safe and effective treatment for humeral simple bone cysts. PMID- 19301803 TI - Variables influencing successful two-incision distal biceps repair. AB - This study elucidated the rate and type of complications after two-incision distal biceps tendon repairs. The study examined the predictive value of patient variables in identifying patients at risk for specific complications, and lastly, it evaluated the placement of the dorsal incision as a predictor of proximal radioulnar synostosis after two-incision repair. A single surgeon's series of 84 consecutive patients who had distal biceps tendon repair using the two-incision, modified Boyd and Anderson approach were retrospectively reviewed. A rate and complication profile was developed. Variable analysis was performed on the study. A second series, a referral group diagnosed with radioulnar synostosis after two incision biceps repair, was also reviewed for comparison. Measurement of nearest distance of dorsal incision to the ulnar crest was made and compared between the study group and the referral group. Six different types of complications occurred with a total of 20 complications. Three complications required repeat operation. No patient variables were found to be statistically significant predictors of complications. The proximity of the dorsal incision to the ulnar crest correlated with the development of radioulnar synostosis. Two-incision distal biceps tendon repair has a low rate of major complications. Patient specific variables do not appear to be associated with the rate or type of complications. Placement of the dorsal incision on the ulnar crest may be associated with the development of radioulnar synostosis. PMID- 19301804 TI - Prosthesis retention, serial debridement, and antibiotic bead use for the treatment of infection following total joint arthroplasty. AB - The clinical outcomes of a consecutive series of deep total joint infections treated with a prosthesis retaining protocol were reviewed. Each patient underwent surgical treatment consisting of serial irrigation and debridements, polyethylene exchange when applicable, interval placement and exchange of intra articular tobramycin/vancomycin-impregnated polymethylmethacrylate beads and 6 weeks of intravenous antibiotics. The primary outcome measures were clinical, radiographic, or laboratory evidence of recurrent prosthetic infection or revision total joint arthroplasty for any reason. At a mean follow-up of 37 months (range, 23-41 months), all infections were eradicated while retaining prosthetic components. No further operative intervention was required for any patient. This treatment protocol can be successful in appropriately selected patients, presenting with early postoperative or acute hematogenous deep total joint infections. PMID- 19301805 TI - The dilemma of the wedding band. AB - The postoperative infection rate in procedures where no ring is worn, and those where a plain metal wedding band is worn under the glove was studied retrospectively. From January 1998 through June 2002, 2127 surgeries were performed by the lead author (D.T.S.), the first 2 years without a wedding band and the next 2 years with a simple platinum wedding band worn under the glove. Attention was paid to sliding the ring proximal and distal on the finger, ensuring scrub solution was under the ring and that the area of skin below the ring was cleansed. Twenty-two postoperative infections were recorded in 2127 surgeries. This is a postoperative infection rate of 1.0%, and <1 (0.449) postoperative infection per month. The "no ring" group totaled 987 cases with an infection rate 1.6%; the "ring" group revealed an infection rate of 0.53% in 1140 cases. Previous studies of jewelry in the operating room do not discuss the type of wedding ring worn, nor do they demonstrate an increased infection rate with wearing jewelry. This study suggests that there is no correlation between wearing a plain wedding band under the surgical glove and an increase in postoperative infections. The crevices and cuticle of the fingers and nails may provide more significant infection risk than a plain metal wedding band. This is a level III retrospective cohort study. PMID- 19301806 TI - Pitfalls of using performance measures to evaluate the quality of hip fracture care. AB - The objective of this study was to determine feasibility of using RAND quality indicators to evaluate hip fracture care. Retrospective chart review was used to determine the adherence to quality indicators and the location of documentation of compliance. A chart abstraction tool was created for systematic extraction of data from multiple chart components. A total of 111 patients underwent operative treatment of a hip fracture and met inclusion criteria in either 1998 or 2003. The main outcome measure was the rate of compliance with quality measures. Overall, compliance was 88% for the 7 performance measures. Physician notes were the most accurate chart component but, if examined alone, would have only resulted in a reported rate of 81% adherence to indicators. Review of the nursing notes, ancillary service notes, results, and orders was required to fully document quality of care. Ceiling effects were noted for 4 of the 7 quality indicators as noncompliance was rare for these measures. The results of this study highlight the need for a thorough method of abstracting multiple chart components to accurately report quality of care. This is an important consideration for any pay-for-performance program. Specifically, the failure to review all chart components may lead to incorrect conclusions about the quality of care delivered by individual providers. In addition, the selection of quality measures subject to ceiling effects may limit the usefulness of quality reporting initiatives. PMID- 19301807 TI - Intramedullary hip screw versus standard compression hip screw: early postoperative rehabilitation comparisons. AB - Studies comparing the intramedullary hip screw and the compression hip screw for fixation of intertrochanteric hip fractures have shown little difference in final functional outcome. However, the characteristics of the rehabilitation process for these implants have not been analyzed. This study used the Functional Independence Measure (FIM Instrument; Uniform Data System for Medical Rehabilitation, Amherst, New York) to better characterize the subtle differences of the perioperative, clinical, and rehabilitative treatment of intertrochanteric fractures using the intramedullary hip screw or compression hip screw. Ninety four patients with isolated intertrochanteric fractures were treated with either an intramedullary hip screw or compression hip screw at our institution. To reduce technical bias, only experienced surgeons were used and patient allocation was surgeon based (eg, surgeons consistently used the same preferred implant). We evaluated the following FIM categories: bed mobility, bed transfer, gait independence, and distance ambulated. Length of stay and level of discharge disposition were also evaluated. The intramedullary hip screw group performed better with bed transfers (P<.05), demonstrated better ambulatory ability at discharge (P<.06), and had an increased gait distance at discharge (P<.07). Skin to-skin operative time and estimated blood loss was significantly less for the intramedullary hip screw group. Length of hospital stay and discharge disposition failed to reach statistical significance. Our study found that when using the FIM scores, some differences were noted in the acute rehabilitation characteristics in patients between the intramedullary hip screw and the compression hip screw. These findings may have medical and social importance as well as significant economic implications. Further study with a larger sample size and more stringent study design are recommended. PMID- 19301808 TI - Deformity correction with an external fixator: ease of use and accuracy? AB - The Taylor spatial frame (Smith & Nephew, Memphis, Tennessee) is a hexapod frame that differs from the well-known Ilizarov fixator in that it works in conjunction with special software programs and can simultaneously correct 6 axes of deformity. The aim of this study was to evaluate the ease of use and accuracy of the Taylor spatial frame. We used the Taylor spatial frame with the older software, Version 2, and the newer Internet-based program, Version 3.1, to achieve our treatment goals in 29 cases. We herein report our results achieved by using the Taylor spatial frame to treat deformity and trauma of the lower limb from November 1999 through May 2005. The hybrid advanced technique of frame application with the use of wires and half-pins was applied in all cases. Included in this study were only those patients whose frames had been removed and whose final results were available. All patients were treated and followed by the senior author (Y.E.). We found that the Taylor spatial frame is easy to apply and that application can be accomplished quickly. The results were clinically and radiographically excellent in all cases at the time of final assessment, and the treatment goals were 100% successful. The only drawbacks we noted are that the Taylor spatial frame is more cumbersome than the conventional Ilizarov frame and the financial cost is comparatively high. PMID- 19301809 TI - Patellofemoral arthroplasty with a custom-fit femoral prosthesis. AB - We reviewed the outcomes of a series of patellar arthroplasty operations with custom-fit femoral prostheses to examine the effectiveness of this procedure in relieving pain and restoring function in the knee. Twenty-two patellofemoral arthroplasty operations were performed in 21 patients (mean age, 48.6 years) at 2 institutions between 1994 and 2002. All patients had advanced patellofemoral arthritis and had undergone an average of 2.5 previous patellofemoral operations. The prosthesis, consisting of a custom-fit chrome cobalt trochlear component and an all-polyethylene patellar button, was implanted in a procedure designed to minimize bone resection. Patients later underwent three-view radiography of the knee to confirm that the prosthesis was positioned correctly. One patient required revision of an undersized patellar button 18 months postoperatively, and 2 other patients had postoperative arthrofibrosis necessitating arthroscopic debridement. No patient required revision of the trochlear component, and no loosening or migration of any component has been found since the first procedure was performed. However, the polyethylene patellar button has worn in 3 patients, and the patella broke in 1 patient. An average of 60 months postoperatively, patients used the Western Ontario and McMaster Universities Osteoarthritis Index to rate their preoperative and present joint pain, stiffness, and function. Patients' mean overall ratings (potential range, 24-96) were significantly lower for their present symptoms (28.4) than for their preoperative symptoms (63.4). Mean scores on each subscale also decreased: from 13.0 to 5.5 for pain, from 5.4 to 2.4 for stiffness, and from 45.0 to 20.6 for function. We conclude that, in carefully selected patients, patellofemoral arthroplasty with a custom-fit prosthesis is a viable surgical treatment for isolated patellofemoral arthritis. PMID- 19301810 TI - Serotonin synthesis in the duodenum controls bone formation. PMID- 19301811 TI - Radiologic case study: Ewing's Sarcoma. AB - Enhance your diagnostic skills with this "test yourself" monthly column, which features a radiograph and challenges you to make a diagnosis. PMID- 19301812 TI - Kinetics of silica nucleation on carboxyl- and amine-terminated surfaces: insights for biomineralization. AB - An in situ, atomic force microscopy- (AFM-)-based experimental approach is developed to directly measure the kinetics of silica nucleation on model biosubstrates under chemical conditions that mimic natural biosilica deposition environments. Relative contributions of thermodynamic and kinetic drivers to surface nucleation are quantified by use of amine-, carboxyl-, and hybrid NH(3)(+)/COO(-)-terminated surfaces as surrogates for charged and ionizable groups on silica-mineralizing organic matrices. The data show that amine terminated surfaces do not promote silica nucleation, whereas carboxyl and hybrid NH(3)(+)/COO(-) substrates are active for silica deposition. The rate of silica nucleation is approximately 18x faster on the hybrid substrates than on carboxylated surfaces, but the free energy barriers to cluster formation are similar on both surface types. These findings suggest that surface nucleation rates are more sensitive to kinetic drivers than previously believed and that cooperative interactions between oppositely charged surface species play important roles in directing the onset of silica nucleation. Further experiments to test the importance of these cooperative interactions with patterned NH(3)(+)/COO(-) substrates, and aminated surfaces with solution-borne anionic species, confirm that silica nucleation is most rapid when oppositely charged species are proximal. By documenting the synergy that occurs between surface groups during silica formation, these findings demonstrate a new type of emergent behavior underlying the ability of self-assembled molecular templates to direct mineral formation. PMID- 19301813 TI - Electron tomography for heterogeneous catalysts and related nanostructured materials. PMID- 19301814 TI - Simultaneous extraction of arsenic and selenium species from rice products by microwave-assisted enzymatic extraction and analysis by ion chromatography inductively coupled plasma-mass spectrometry. AB - A microwave-assisted enzymatic extraction (MAEE) method was developed for the simultaneous extraction of arsenic (As) and selenium (Se) species in rice products. The total arsenic and selenium content in the enzymatic extracts were determined by inductively coupled plasma mass spectrometry (ICP-MS), while the speciation analysis was performed by ion chromatography coupled to inductively coupled plasma-mass spectrometry (IC-ICP-MS). The main factors affecting the enzymatic extraction process were evaluated in NIST SRM-1568a rice flour. The optimum extraction conditions were 500 mg of sample, 50 mg of protease XIV, and 25 mg of alpha-amylase in aqueous medium during 40 min at 37 degrees C. The extraction recoveries of total As and Se reached 100 +/- 3 and 80 +/- 4%, respectively. The species stability study during the MAEE process did not show transformation of the target species in rice products. The results of As speciation obtained for SRM-1568a were in agreement with previous studies of As speciation performed on the same reference material. The proposed method was applied to the determination of As and Se species in rice and rice-based cereals. Arsenite [As(III)], arsenate [As(V)], dimethylarsinic acid (DMA), and selenomethionine (SeMet) were the predominant species identified in rice products. PMID- 19301815 TI - Occurrence of deoxynivalenol and its major conjugate, deoxynivalenol-3-glucoside, in beer and some brewing intermediates. AB - Since deoxynivalenol (DON), the main representative of Fusarium toxic secondary metabolites, is a relatively common natural contaminant in barley, its traces can be detected in many commercial beers. Our previous study reporting for the first time the occurrence of relatively high levels of DON-3-glucoside (DON-3-Glc) in malt and beer prepared from relatively "clean" barley (semiscale experimental conditions) induced a follow-up investigation focused on this DON conjugate in commercial beers. The current survey involving in total 176 beers, representing different brands, and collected at various markets, has documented a ubiquitous occurrence of DON-3-Glc in this product. Its levels even exceeded that of free DON in some samples; the highest level found was 37 microg/L. In addition to glucosylated DON, its acetylated forms (ADONs) were also common contaminants in most of the beers. Generally, stronger beers (higher alcohol content) tended to contain higher levels of DON and its conjugates. No distinct relationship between the contamination of malt and beer was observed in samples collected from several breweries. Attention was also paid to comparison of data on malts obtained by LC MS/MS and ELISA DON-dedicated kits. The latter provided apparently higher levels of DON, the most distinct difference being observed for malts processed at higher temperatures (caramel and roasted malts). The nature of this phenomenon has not yet been explained; in addition to cross-reacting species, other factors, such as the higher content of dark pigment, can also be the cause. PMID- 19301816 TI - Role of the hydrogen bonding heteroatom-Lys53 interaction between the p38alpha mitogen-activated protein (MAP) kinase and pyridinyl-substituted 5-membered heterocyclic ring inhibitors. AB - In the framework of investigating the role of heteroatoms in pyridinyl substituted 5-membered (hetero)cycles as potential p38alpha MAP kinase inhibitor scaffolds, cyclopentene, pyrrole, furan, and imidazole analogues were synthesized and tested with respect to their ability to inhibit p38alpha MAP kinase. The vicinal pyridine/4-fluorophenyl pharmacophore was conserved, such as in the prototypical imidazole inhibitor SB203580. The strength of the HB interaction was calculated and compared to the biological data. PMID- 19301817 TI - Hibiscus sabdariffa inhibits vascular smooth muscle cell proliferation and migration induced by high glucose--a mechanism involves connective tissue growth factor signals. AB - Recently, the herbal extract of Hibiscus sabdariffa was shown to have multiple bioactive effects, including anti-atherosclerosis. On the basis of this, we aimed to examine whether the polyphenolic isolate of H. sabdariffa (HPI) could protect high-glucose-treated vascular smooth muscle cell (VSMC) and its putative transduction signals. Results showed that HPI dose- and time-dependently reduced the high-glucose-stimulated cell proliferation and migration. HPI suppressed the proliferating cell nuclear antigen (PCNA) level and matrix metalloproteinase (MMP)-2 activation. In addition, the expressions of connective tissue growth factor (CTGF) and receptor of advanced glycation end product (RAGE) enhanced by high glucose were prominently suppressed by HPI. The proliferation signal mediated by high glucose was demonstrated via CTGF/RAGE, while MMP-2 was regulated by CTGF but not RAGE. Conclusively, the results suggest that HPI potentially can be a promising adjuvant herbal therapy for diabetic patients. PMID- 19301818 TI - Hpf2 glycan structure is critical for protection against protein haze formation in white wine. AB - Grape-derived proteins can form haze in wine. Some cell-wall glycoproteins of Saccharomyces cerevisiae are capable of reducing protein haze formation. The basis of their haze protective activity is not yet understood. One of the S. cerevisiae cell-wall proteins, Hpf2, was produced in Pichia pastoris . An altered glycan structure in the P. pastoris -produced protein was associated with decreased solubility in water and reduced capacity to mitigate haze formation compared to native Hpf2 protein from S. cerevisiae. alpha-1,2-Linked mannose in the glycan chain was shown to be required for haze protective activity using a series of S. cerevisiae deletion mutants (mnn1-Delta, mnn2-Delta, mnn4-Delta, and mnn5-Delta), defective in different aspects of glycan processing. The effect of media additives phthalate, casamino acids, and yeast nitrogen base on Hpf2 production in P. pastoris were also evaluated. Casamino acids were shown to suppress Hpf2 production in P. pastoris . PMID- 19301819 TI - Quantitative lipid analysis and life span of the fat-3 mutant of Caenorhabditis elegans. AB - The long-chain polyunsaturated fatty acids (LC PUFAs) docosahexaenoic acid (DHA, 22:6n3) and eicosapentaenoic acid (EPA, 20:5n3) are important for health and development of organisms, but the precise biological function of these molecules is not known. It has been suggested that they may play a part in aging, as they are highly susceptible to oxidation. A genetic mutant of Caenorhabditis elegans (fat-3), which lacks a functional delta-6 desaturase, and thus LC PUFAs including EPA, allows dietary manipulation of long-chain n3 fatty acids in this nematode. The life span of C. elegans strains N2 (wild-type) and BX30 [fat-3(wa22)] with and without supplemental EPA and DHA was analyzed. In addition, quantitative analysis was performed on total lipids, phospholipids, and triglycerides, as it is important to understand where fatty acids are being partitioned among the various lipid classes. The results show a beneficial effect of these molecules on the life span of C. elegans and will aid in the elucidation of the underlying causes of PUFA deficiency in the simple animal C. elegans as well as in humans. PMID- 19301820 TI - Cross coupling reactions of chiral secondary organoboronic esters with retention of configuration. AB - We report the first example of a coupling reaction of chiral secondary boronic esters generated by the hydroboration of vinyl arenes. In order for the reaction to take place in high yields, the use of silver oxide as a base and the presence of at least 8 equiv of triphenyl phosphine per Pd are required. The reaction proceeds with >90% retention of configuration in all cases except one. Remarkably, the linear boronate ester does not react under these conditions. PMID- 19301821 TI - Pyrido[2,3-e]-1,2,4-triazolo[4,3-a]pyrazin-1-one as a new scaffold to develop potent and selective human A3 adenosine receptor antagonists. Synthesis, pharmacological evaluation, and ligand-receptor modeling studies. AB - The paper describes a new class of human (h) A(3) adenosine receptor antagonists, the 2-arylpyrido[2,3-e]-1,2,4-triazolo[4,3-a]pyrazin-1-one derivatives (PTP), either 4-oxo (1-6, series A) or 4-amino-substituted (7-20, series B). In both series A and B, substituents able to act as hydrogen bond acceptors (OMe, OH, F, COOEt) were inserted on the 2-phenyl ring. In series B, cycloalkyl and acyl residues were introduced on the 4-amino group. Some of the new derivatives showed high hA(3) AR affinities (K(i) < 50 nM) and selectivities vs both hA(1) and hA(2A) receptors. The selected 4-benzoylamino-2-(4-methoxyphenyl)pyrido[2,3-e] 1,2,4-triazolo[4,3-a]pyrazin-1-one (18), tested in an in vitro rat model of cerebral ischemia, proved to be effective in preventing the failure of synaptic activity induced by oxygen and glucose deprivation in the hippocampus. Molecular docking of this new class of hA(3) AR antagonists was carried out to depict their hypothetical binding mode to our refined model of hA(3) receptor. PMID- 19301822 TI - Design, synthesis, and evaluation of novel fluoroquinolone-aminoglycoside hybrid antibiotics. AB - A series of new hybrid structures containing fluoroquinolone (ciprofloxacin) and aminoglycoside (neomycin) antibiotics linked via 1,2,3-triazole moiety were designed and synthesized, and their antibacterial activities were determined against both Gram-negative and Gram-positive bacteria, including resistant strains. The nature of spacers in both the ciprofloxacin and neomycin parts greatly influenced the antibacterial activity. The majority of hybrids was significantly more potent than the parent neomycin and overcame most prevalent types of resistance associated with aminoglycosides. Selected hybrids inhibited bacterial protein synthesis with the potencies similar to or better than that of neomycin and were up to 32-fold more potent inhibitors than ciprofloxacin for the fluoroquinolone targets, DNA gyrase and toposiomerase IV, indicating a balanced dual mode of action. Significant delay of resistance formation was observed in both E. coli and B. subtilis to the treatment with ciprofloxacin-neomycin hybrid in comparison to that of each drug separately or their 1:1 mixture. PMID- 19301823 TI - Shape and size control of oriented polyaniline microstructure by a self-assembly method. AB - Oriented polyaniline (PANI) microstructures (e.g., bulks, spheres, flakes, and fibers) were prepared without a template by polymerization at 80 degrees C and crystal growth at 0 degrees C and using a self-assembly method in the presence of hydrochloric acid (HCl) as the dopant. It was found that the shape and size of the resulting PANI microstructures depended on the HCl dosage and aniline concentration. Polyanilinium salt precipitates, as a templete, drive the formation of highly oriented PANI microstructures. The infrared and UV-vis absorption spectra and X-ray diffraction were used to characterize the molecular structures of the PANI microstructures. The results showed that their main structure was identical to those of the emeraldine salt form of PANI. PMID- 19301824 TI - Formation of monodisperse FePt alloy nanocrystals using air-stable precursors: fatty acids as alloying mediator and reductant for Fe3+ precursors. AB - High quality FePt nanocrystals were synthesized by a simple and high performance synthetic method in a hydrocarbon solvent. Instead of a toxic and dangerous Fe(CO)(5) precursor, air-stable and generic inorganic and organic iron salts were used as the iron precursors. Fatty acid molecules were identified as both efficient reducing reagents for the Fe(3+) precursors and as an alloying mediator. The alloying process occurred in a two-step fashion. The Pt nanocrystal seeds initially formed at relatively low temperatures, with the need to be stabilized by a small amount of amines. Conversely, the final FePt nanocrystals could be stabilized with fatty acids as the main ligands. The mechanisms revealed here not only help to understand the formation of alloy nanocrystals but also shed new light on the formation of transition metal oxide nanocrystals using metal fatty acid salts as the precursors. PMID- 19301825 TI - Quinone-enhanced reduction of nitric oxide by xanthine/xanthine oxidase. AB - The quinones 1,4-naphthoquinone, methyl-1,4-naphthoquinone, tetramethyl-1,4 benzoquinone, 2,3-dimethoxy-5-methyl-1,4-benzoquinone, 2,6-dimethylbenzoquinone, 2,6-dimethoxybenzoquinone, and 9,10-phenanthraquinone enhance the rate of nitric oxide reduction by xanthine/xanthine oxidase in nitrogen-saturated phosphate buffer (pH 7.4). Maximum initial rates of NO reduction (V(max)) and the amount of nitrous oxide produced after 5 min of reaction increase with quinone one- and two electron redox potentials measured in acetonitrile. One of the most active quinones of those studied is 9,10-phenanthraquinone with a V(max) value 10 times larger than that corresponding to the absence of quinone, under the conditions of this work. Because NO production is enhanced under hypoxia and under certain pathological conditions, the observations obtained in this work are very relevant to such conditions. PMID- 19301826 TI - Inherent stereospecificity in the reaction of aflatoxin B(1) 8,9-epoxide with deoxyguanosine and efficiency of DNA catalysis. AB - Kinetic analysis of guanine alkylation by aflatoxin B(1) exo-8,9-epoxide, the reactive form of the hepatocarcinogen aflatoxin B(1), shows the reaction to be >2000 times more efficient in DNA than in aqueous solution, that is, with free 2' deoxyguanosine. Thermodynamic analysis reveals AFB(1) exo-8,9-epoxide intercalation as the predominant source of the observed DNA catalytic effect. However, the known exo > endo epoxide stereospecificity of the DNA alkylation is observed even with free deoxyguanosine (ratio >20:1 determined by LC-MS and NMR measurements), as predicted by theoretical calculations [ Bren , U. , et al. ( 2007 ) Chem. Res. Toxciol. 20 , 1134 - 1140 ]. PMID- 19301827 TI - Chemoinformatic analysis of combinatorial libraries, drugs, natural products, and molecular libraries small molecule repository. AB - A multiple criteria approach is presented, that is used to perform a comparative analysis of four recently developed combinatorial libraries to drugs, Molecular Libraries Small Molecule Repository (MLSMR) and natural products. The compound databases were assessed in terms of physicochemical properties, scaffolds, and fingerprints. The approach enables the analysis of property space coverage, degree of overlap between collections, scaffold and structural diversity, and overall structural novelty. The degree of overlap between combinatorial libraries and drugs was assessed using the R-NN curve methodology, which measures the density of chemical space around a query molecule embedded in the chemical space of a target collection. The combinatorial libraries studied in this work exhibit scaffolds that were not observed in the drug, MLSMR, and natural products databases. The fingerprint-based comparisons indicate that these combinatorial libraries are structurally different than current drugs. The R-NN curve methodology revealed that a proportion of molecules in the combinatorial libraries is located within the property space of the drugs. However, the R-NN analysis also showed that there are a significant number of molecules in several combinatorial libraries that are located in sparse regions of the drug space. PMID- 19301828 TI - Effect of serum on an RNA aptamer-based electrochemical sensor for theophylline. AB - Electrochemical performance of the ferrocene (Fc) redox-labeled RNA aptamer based sensor for theophylline (Th) is essentially inhibited in serum, but is restored in serum-free buffer solutions. This phenomenon is inconsistent with the data on methylene-blue-labeled aptamer beacon systems, which operational potential window is more negative compared to the Fc redox label. Electrochemical studies with a ferricyanide redox probe, having redox potential close to the Fc redox couple, and interfacial capacitance measurements unambiguously demonstrate that it is adsorption of serum proteins at positively charged electrode surface that slows down the kinetics of the electrode reactions in serum and interferes with the biosensor performance. In filtered serum solutions, in the absence of serum proteins, the Fc-labeled aptamer-based biosensor performed similarly to the pure buffer solutions, ad the signal for Th could be linearly calibrated versus Th concentration. These results on interfacial effects of serum are of particular importance for future research and development of the beacon-type biosensors for in vivo applications. PMID- 19301829 TI - Transition force measurement between two negligibly charged surfaces: a new perspective on nanoparticle halos. AB - Since 2001, silica microspheres have been reported to be stabilized by highly charged hydrous ZrO(2) nanoparticles which form halos around the microspheres at pH 1.5. However, the exact mechanisms behind this novel stabilization method in terms of the relevant interaction forces remain unclear. In order to gain a greater insight into this mechanism, the interaction between a silica flat and a silica sphere in different ZrO(2) nanoparticle suspensions was investigated by the colloid probe technique. The interaction force between a silica flat and a 600 nm silica sphere was first investigated in a ZrO(2) nanoparticle (D approximately 8 nm) suspension with volume fractions of 10(-3), 10(-4), 10(-5), and 10(-6). When the volume fraction of ZrO(2) is 10(-6), only a purely attractive van der Waals force was observed between the silica surfaces. With an increase in the ZrO(2) nanoparticle volume fraction, a peak was detected on the transition force curve at a ZrO(2) volume fraction of 10(-5) while a purely repulsion force was observed for ZrO(2) volume fractions of 10(-4) and 10(-3). The average distance difference between the peak and the zero distance point on the transition force curve which should define the distance between the halo on the microsphere is approximately 2.3 nm. Additionally, the repulsion increases with the effective zeta potential of the binary composite sphere (BCS, the entity of the silica sphere and the surrounding zirconia particles) on an increase of the nanoparticle volume fraction while the adhesion force decreases, which indicates a denser nanoparticle halo. PMID- 19301830 TI - Fluorescence microscopy imaging of giant folding in a catanionic monolayer. AB - The behavior of the catanionic system of dioctadecyldimethylammonium bromide (DODAB) and sodium dodecyl sulfate (SDS) was investigated at 23 +/- 1 degrees C at the air-water interface using a Langmuir trough. The surface pressure as a function of surface area was measured while monitoring domain structures using epifluorescence microscopy. At high surface densities, the monolayer exhibits collapse through reversible folding at about 47 mN m(-1). This corresponds to the DODAB collapse surface pressure. The number of folds increases with the rate of compression speed and is history-dependent. PMID- 19301831 TI - Theoretical study of the effective Chemical Shielding Anisotropy (CSA) in peptide backbone, rating the impact of CSAs on the cross-correlated relaxations in L alanyl-L-alanine. AB - The dependence of the effective chemical shielding anisotropy (effective CSA, Deltasigma(eff)) on the phi and psi peptide backbone torsion angles was calculated in the l-alanyl-l-alanine (LALA) peptide using the DFT method. The effects of backbone conformation, molecular charge including the cation, zwitterion, and anion forms of the LALA peptide, and the scaling taking into account the length of the dipolar vector were calculated for the effective CSAs in order to assess their structural behaviors and to predict their magnitudes which can be probed for the beta-sheet and alpha-helix backbone conformations via measurement of the cross-correlated relaxation rates (CCR rates). Twenty different CSA-DD cross-correlation mechanisms involving the amide nitrogen and carbonyl carbon chemical shielding tensors and the C(alpha)H(alpha) (alpha-carbon group), NH(N) (amide group), C(alpha)H(N), NH(alpha), C'H(alpha), and C'H(N) (alpha = alpha1, alpha2) dipolar vectors were investigated. The X C(alpha)H(alpha) (X = N, C'; alpha = alpha1, alpha2) cross-correlations, which had already been studied experimentally, exhibited overall best performance of the calculated effective CSAs in the LALA molecule; they spanned the largest range of values upon variation of the psi and phi torsions and depended dominantly on only one of the two backbone torsion angles. The X-NH(N) (X = N, C') cross-correlations, which had been also probed experimentally, depended on both backbone torsions, which makes their structural assignment more difficult. The N-NH(alpha2) and N-C'H(alpha1) cross-correlations were found to be promising for the determination of various backbone conformations due to the large calculated range of the scaled effective CSA values and due to their predominant dependence on the psi and phi torsions, respectively. The 20 calculated dependencies of effective CSAs on the two backbone torsion angles can facilitate the structural interpretation of CCR rates. PMID- 19301832 TI - Efficient escape from endosomes determines the superior efficiency of multicomponent lipoplexes. AB - Designer multicomponent lipoplexes have recently emerged as especially promising transfection candidates, since they are from 10 to 100 times more efficient than binary complexes usually employed for gene delivery purposes. Here, we show, for the first time, that after internalization binary complexes of lower transfection potency remain in compact perinuclear endosomes, while multicomponent systems have intrinsic endosomal rupture properties that allow plasmid DNA to escape from endosomes with extremely high efficiency. Endosomal rupture results in an extraordinarily homogeneous distribution of unbound plasmid DNA throughout the cytoplasm and in the nucleus. PMID- 19301833 TI - Monitoring the interaction of a single G-protein key binding site with rhodopsin disk membranes upon light activation. AB - Heterotrimeric G-proteins interact with their G-protein-coupled receptors (GPCRs) via key binding elements comprising the receptor-specific C-terminal segment of the alpha-subunit and the lipid anchors at the alpha-subunit N-terminus and the gamma-subunit C-terminus. Direct information about diffusion and interaction of GPCRs and their G-proteins is mandatory for an understanding of the signal transduction mechanism. By using single-particle tracking, we show that the encounters of the alpha-subunit C-terminus with the GPCR rhodopsin change after receptor activation. Slow as well as less restricted diffusion compared to the inactive state within domains 60-280 nm in length was found for the receptor bound C-terminus, indicating short-range order in rhodopsin packing. PMID- 19301834 TI - Reinvestigation of the branching ratio of the CN + O2 reaction. AB - The reaction of the CN radical with O(2) was studied using infrared diode laser absorption spectroscopy. Detection of NO and secondary N(2)O products was used to directly measure the product branching ratio. After consideration of possible secondary chemistry and comparison to kinetic modeling simulations, the branching ratio of the CN + O(2) reaction into the NO + CO channel was determined to be phi (NO + CO) = 0.20 +/- 0.02, with little or no temperature dependence over the range 296-475 K. PMID- 19301835 TI - Size-tunable trehalose-based nanocavities: synthesis, structure, and inclusion properties of large-ring cyclotrehalans. AB - An efficient strategy toward the synthesis of large-ring cyclodextrin (CD) analogs alternating alpha,alpha'-trehalose disaccharide subunits and pseudoamide segments (cyclotrehalans, CTs), involving a bimolecular macrocyclization reaction as the key step, is reported. NMR and molecular modeling confirmed that the eight and ten alpha-d-glucopyranoside subunits in tetrameric and pentameric CT homologues (CT4 and CT5, respectively) are magnetically equivalent, as in the gamma and epsilonCD counterparts. Yet, the orientation of the monosaccharide constituents is reversed in CTs as compared with CDs, the beta-face being directed to the inside of the nanometric cavity while the alpha-face remains in contact with the bulk solvent. Molecular mechanics and dynamics experiments revealed that the cyclooligosaccharide architecture in CT4 and CT5 is relatively flexible, which is in contrast to that previously observed for the first members of the CT series (CT2 and CT3 oligomers). Thus, although in their fully expanded conformation their cavity size is close to that of gammaCD, the higher mobility of the pseudoamide bridges as compared with classical glycosidic linkages endows these hosts with induced fitting capabilities toward smaller guests. PMID- 19301836 TI - Mechanism of Cdc25B phosphatase with the small molecule substrate p-nitrophenyl phosphate from QM/MM-MFEP calculations. AB - Cdc25B is a dual-specificity phosphatase that catalyzes the dephosphorylation of the Cdk2/CycA protein complex. This enzyme is an important regulator of the human cell cycle and has been identified as a potential anticancer target. In general, protein tyrosine phosphatases are thought to bind the dianionic form of the phosphate and employ general acid catalysis via the Asp residue in the highly conserved WPD-loop. However, the Cdc25 phosphatases form a special subfamily based on their distinct differences from other protein tyrosine phosphatases. Although Cdc25B contains the (H/V)CX(5)R catalytic motif present in all other protein tyrosine phosphatases, it lacks an analogous catalytic acid residue. No crystallographic data currently exist for the complex of Cdc25B with Cdk2/CycA, so in addition to its natural protein substrate, experimental and theoretical studies are often carried out with small molecule substrates. In an effort to gain understanding of the dephosphorylation mechanism of Cdc25B with a commonly used small molecule substrate, we have performed simulations of the rate-limiting step of the reaction catalyzed by Cdc25B with the substrate p-nitrophenyl phosphate using the recently developed QM/MM Minimum Free Energy Path method (Hu et al. J. Chem. Phys. 2008, 034105). We have simulated the first step of the reaction with both the monoanionic and the dianionic forms of the substrate, and our calculations favor a mechanism involving the monoanionic form. Thus, Cdc25 may employ a unique dephosphorylation mechanism among protein tyrosine phosphatases, at least in the case of the small molecule substrate p-nitrophenyl phosphate. PMID- 19301837 TI - Phase transitions in the crystals of L- and DL-cysteine on cooling: the role of the hydrogen-bond distortions and the side-chain motions. 2. DL-cysteine. AB - Structural strain and a first-order phase transition in the crystalline DL cysteine on cooling and on reverse heating were followed by Raman spectroscopy and X-ray diffraction. The transition is reversible and has a large hysteresis (over 100 K). The temperature at which the transition is observed depends strongly on the cooling/heating rate. The phase transition is accompanied by crystal fragmentation. The low-temperature phase could be obtained not only as a result of the solid-state transformation in situ as a polycrystalline sample (with strong preferred orientation, or without it, depending on the preparative technique), but also (using an original crystallization technique) as a single crystal of the quality suitable for structural analysis. For the first time, the crystal structure of the low-temperature phase was solved independently by powder and by single-crystal diffraction techniques. The spectral changes were correlated with the precise diffraction data on the intramolecular conformations and the intermolecular hydrogen bonding before and after the phase transition. The role of the distortion of the intermolecular hydrogen bonds and of the motions of the -CH(2)SH side chains in the phase transition is discussed in a comparison with the low-temperature phase transition in L-cysteine, which is of a different type and preserves the single crystals intact (Kolesov et al. J. Phys. Chem. B, 2008, 112 (40), 12827-12839). PMID- 19301838 TI - Dye-adsorption-induced gelation of suspensions of spherical and rodlike zinc oxide nanoparticles in organic solvents. AB - The adsorption of amphiphilic Ru(II) complex Z907 onto the surface of ZnO nanospheres and nanorods causes the gelation of organic solvents, such as THF and acetone. The gels are thermally stable at very low concentration (nanoparticle volume fraction phi = 0.009) but mechanically fragile, with the behavior being dependent on the nature of the solvent, nanoparticle concentration, and the Z907/ZnO mole/weight ratio. Rheological experiments confirmed that the solid component built up a network to give a viscoelastic gel-phase material with a weak value of storage modulus G'. However, TEM and SEM experiments did not give evidence that nanoparticle long-range ordering occurred under the experimental conditions investigated. Moreover, time-dependent SAXS measurements pointed to a decrease in the nanoparticle aggregate size upon gelation. All together, the data obtained might be rationalized in terms of the aggregate-to-aggregate transition in solution, with the primitive large aggregates giving rise to smaller ones upon reaction with Z907. The resulting smaller hybrid aggregates could be the active species that act as self-assembling components in the gelation process. Given the interesting electronic and photonic properties of zinc oxide nanoparticles, such hybrid organic-inorganic gels could open new directions in materials science, low cost electronics, and photovoltaics. PMID- 19301839 TI - Submillimeter-scale surface gradients of immobilized protein ligands. AB - We describe a method to produce antibody-captured ligand gradients over biologically relevant distances (hundreds of micrometers) whereby the ligand density and gradient shape may be tailored. Separation of the ligand from the solid-phase surface ensures that the biological activity of the ligand remains unaffected by immobilization. Our method involves the use of a plasma-masking method to generate a surface chemical gradient on a glass substrate to which the 9E10 antibody is covalently coupled. This antibody captures myc-tagged biomolecules. In our example, the antibody is then used to immobilize a gradient of the intercellular signaling molecule delta-like-1 (Dll1). To visualize the gradient of Dll1, we have used the multistep approach of binding with rabbit anti Dll1 primary antibody and then adding colloidal-gold-conjugated secondary antibody. PMID- 19301840 TI - TiO2-catalyzed photodegradation of porphyrins: mechanistic studies and application in monolayer photolithography. AB - Patterned mixed monolayers of porphyrins on nanocrystalline TiO(2) films were fabricated by substrate-catalyzed monolayer photolithography. Tin(IV) protoporphyrin IX (SnPP), zinc(II) protoporphyrin IX (ZnPP), and iron(III) meso tetra(4-carboxyphenyl)porphine (FeTCP) were adsorbed to TiO(2) through the carboxyl groups, yielding saturation surface amounts per projected area of approximately 10(-7) mol/cm(2). Illumination of SnPP- and ZnPP-functionalized TiO(2) films with 355 nm light caused the desorption of the porphyrins, most likely through oxidative decarboxylation. SnPP was removed more rapidly than ZnPP. The faster kinetics was due, in part, to the contribution of other photochemical pathways including TiO(2)-catalyzed photoreduction and direct photodegradation reactions. Patterned binary monolayers were prepared by the photoinduced desorption of a protoporphyrin, followed by the adsorption of FeTCP to previously illuminated regions of the surface. PMID- 19301841 TI - Solvo-hydrothermal approach for the shape-selective synthesis of vanadium oxide nanocrystals and their characterization. AB - A new solvo-hydrothermal method has been developed for the synthesis of uniform vanadium oxide nanocrystals (NCs) with various sizes and shapes in aliphatic amine/toluene/water using V(V) diperoxo alkylammonium complexes. The vanadium complex precursors were prepared from an ion exchange reaction of V(V) diperoxo gels and tetraalkylammonium bromide in the water-toluene mixture using H(2)O(2) solution and commercial bulk V(2)O(5) powders as starting vanadium gel source. The obtained VO(2) NC products were characterized by means of transmission electron microscopy (TEM), selected area electron diffraction (SAED), scanning electron microscopy (SEM), powder X-ray diffraction (XRD), X-ray photoelectron spectra (XPS), Fourier transform infrared absorption spectroscopy (FTIR), thermogravimetric differential thermal analysis (TGA-DTA), and nitrogen adsorption/desorption (BET). The size and shape of NCs can be controlled by different synthesis parameters such as water content, steric ligands of complexes, alkyl chain lengths of capping aliphatic amines, as well as nature of solvent. Monodisperse vanadium oxide NCs with various sizes and shapes, nanospheres, nanocubes, nanorices, and nanorods, can be easily achieved. The possible mechanisms for the formation of vanadium complex precursors and vanadium oxide NCs as well as the shape evolution of NCs were also discussed. The as-made vanadium oxide products exhibited the monoclinic rutile VO(2) structure, which was however converted to the orthorhombic V(2)O(4.6) structure after calcination in air. The XPS results also revealed only one V(4+) state for the as-made sample; however, the coexistence of V(5+) and V(4+) states and two components of oxygen associated with OV and O-V for the calcined samples on the vanadium oxide NC surface were observed. The surface chemical composition of both as-made and calcined samples were found to be VO(2) and V(2)O(5-x) (x = 0.4), respectively. Our approach may provide a novel route for the extended synthesis to other inorganic NCs. PMID- 19301842 TI - Formation and self-organization kinetics of alpha-CD/PEO-based pseudo polyrotaxanes in water. A specific behavior at 30 degrees C. AB - alpha-Cyclodextrins (alpha-CDs) have the ability to form inclusion complexes with poly(ethylene oxide) (PEO) polymer chains. These pseudo-polyrotaxanes (PPRs) can be obtained by quenching an alpha-CD/PEO mixture in water from 70 degrees C down to a lower temperature (typically in the range from 5 to 30 degrees C) thanks to favorable interactions between alpha-CD cavities and PEO chains. Moreover, starting from a liquid alpha-CD/PEO mixture at a total mass fraction of 15% w/w at 70 degrees C, the formation of PPRs with time at a lower temperature induces a white physical gel with time, and phase separation is observed. We established that PPR molecules are exclusively found in the precipitated phase although unthreaded alpha-CD molecules and unthreaded PEO chains are in the liquid phase. At 30 degrees C, the physical gel formation is much slower than at 5 degrees C. At 30 degrees C, we established that, in a first step, alpha-CDs thread onto PEO chains, forming PPR molecules which are not in good solvent conditions in water. At a higher length scale, rapid aggregation of the PPR molecules occurs, and threaded alpha-CD-based nanocylinders form (cylinder length L = 5.7 nm and cylinder radius R = 4.7 nm). At a higher length scale, alpha-CD-based nanocylinders associate in a Gaussian way, engendering the formation of precipitated domains which are responsible for the high turbidity of the studied system. At the end of this first step (i.e., after 20 min), the system still remains liquid and the PPRs are totally formed. Then, in a second step (i.e., after 150 min), the system undergoes its reorganization characterized by a compacity increase of the precipitated domains and forms a physical gel. We found that PPRs are totally formed after 20 min at 30 degrees C and that the system stays in a nongel state up to 150 min. This opens new perspectives regarding the PPR chemical modification: between these two characteristic times, we can easily envisage an efficient chemical modification of the PPR molecules in water, as for instance an end-capping reaction leading to the synthesis of polyrotaxanes. PMID- 19301844 TI - Electrokinetic transport in microchannels with random roughness. AB - We present a numerical framework to model the electrokinetic transport in microchannels with random roughness. The three-dimensional microstructure of the rough channel is generated by a random generation-growth method with three statistical parameters to control the number density, the total volume fraction, and the anisotropy characteristics of roughness elements. The governing equations for the electrokinetic transport are solved by a high-efficiency lattice Poisson Boltzmann method in complex geometries. The effects from the geometric characteristics of roughness on the electrokinetic transport in microchannels are therefore modeled and analyzed. For a given total roughness volume fraction, a higher number density leads to a lower fluctuation because of the random factors. The electroosmotic flow rate increases with the roughness number density nearly logarithmically for a given volume fraction of roughness but decreases with the volume fraction for a given roughness number density. When both the volume fraction and the number density of roughness are given, the electroosmotic flow rate is enhanced by the increase of the characteristic length along the external electric field direction but is reduced by that in the direction across the channel. For a given microstructure of the rough microchannel, the electroosmotic flow rate decreases with the Debye length. It is found that the shape resistance of roughness is responsible for the flow rate reduction in the rough channel compared to the smooth channel even for very thin double layers, and hence plays an important role in microchannel electroosmotic flows. PMID- 19301845 TI - Highly sensitive immunoassay of lung cancer marker carcinoembryonic antigen using surface-enhanced Raman scattering of hollow gold nanospheres. AB - A quick and reproducible surface-enhanced Raman scattering (SERS)-based immunoassay technique, using hollow gold nanospheres (HGNs) and magnetic beads, has been developed. Here, HGNs show strong enhancement effects from individual particles because hot spots can be localized on the pinholes in the hollow particle structure. Thus, HGNs can be used for highly reproducible immunoanalysis of cancer markers. Magnetic beads were used as supporting substrates for the formation of the immunocomplex. This SERS-based immunoassay technique overcomes the problem of slow immunoreaction caused by the diffusion-limited kinetics on a solid substrate because all of the reactions occur in solution. For the validation of our SERS immunoassay, a well-known lung cancer marker, carcinoembryonic antigen (CEA), was used as a target marker. According to our experimental results, the limit of detection (LOD) was determined to be 1-10 pg/mL, this value being about 100-1000 times more sensitive than the LOD of enzyme-linked immunosorbent assay. Furthermore, the assay time took less than 1 h, including washing and optical detection steps. PMID- 19301846 TI - Detection of carbendazim by surface-enhanced Raman scattering using cyclodextrin inclusion complexes on gold nanorods. AB - Surface-enhanced Raman scattering (SERS) was used for the detection of carbendazim using gold nanorods derivatized with a beta-cyclodextrin derivative to bind this fungicide. Gold nanorods were synthesized with an aspect ratio of 3.3 to match the 785 nm excitation wavelength used in the SERS studies. A thiolated cyclodextrin-based sensor molecule was synthesized, and its inclusion complex with carbendazim was formed and studied using SERS spectroscopy. SERS analysis of the inclusion complex at different concentrations in the presence of gold nanorods afforded good quality Raman spectra of carbendazim at micromolar concentrations. Quantitative analysis was preformed using partial least-squares (PLS), and a calibration plot for these data was generated. Results suggest that carbendazim concentrations as low as 50 microM can be accurately detected using the described SERS assay. PMID- 19301847 TI - Thermodynamic reciprocity of the inhibitor binding to the active site and the interface binding region of IB phospholipase A2. AB - Interfacial activation of pig pancreatic IB phospholipase A(2) (PLA2) is modeled in terms of the three discrete premicellar complexes (E(i)(#), i = 1, 2, or 3) consecutively formed by the cooperative binding of a monodisperse amphiphile to the i-face (the interface binding region of the enzyme) without or with an occupied active site. Monodisperse PCU, the sn-2-amide analogue of the zwitterionic substrate, is a competitive inhibitor. PCU cooperatively binds to the i-face to form premicellar complexes (E(i), i = 1 or 2) and also binds to the active site of the premicellar complexes in the presence of calcium. In the E(i)I complex formed in the presence of PCU and calcium, one inhibitor molecule is bound to the active site and a number of others are bound to the i-face. The properties of the E(i) complexes with PCU are qualitatively similar to those of E(i)(#) formed with decylsulfate. Decylsulfate binds to the i-face but does not bind to the active site in the presence of calcium, nor does it interfere with the binding of PCU to the active site in the premicellar complexes. Due to the strong coupling between binding at the i-face and at the active site, it is difficult to estimate the primary binding constants for each site in these complexes. A model is developed that incorporates the above boundary conditions in relation to a detailed balance between the complexes. A key result is that a modest effect on cooperative amphiphile binding corresponds to a large change in the affinity of the inhibitor for the active site. We suggest that besides the binding to the active site, PCU also binds to another site and that full activation requires additional amphiphiles on the i-face. Thus, the activation of the inhibitor binding to the active site of the E(2)(#) complex or, equivalently, the shift in the E(1)(#) to E(2)(#) equilibrium by the inhibitor is analogous to the allosteric activation of the substrate binding to the enzyme bound to the interface. PMID- 19301848 TI - Allosteric effect of amphiphile binding to phospholipase A(2). AB - In the preceding paper, we showed that the formation of the second premicellar complex of pig pancreatic IB phospholipase A2 (PLA2) can be considered a proxy for interface-activated substrate binding. Here we show that this conclusion is supported by results from premicellar E(i)(#) (i = 1, 2, or 3) complexes with a wide range of mutants of PLA2. Results also show a structural basis for the correlated functional changes during the formation of E(2)(#), and this is interpreted as an allosteric T (inactive) to R (active) transition. For example, the dissociation constant K(2)(#) for decylsulfate bound to E(2)(#) is lower at lower pH, at higher calcium concentrations, or with an inhibitor bound to the active site. Also, the lower limits of the K(2)(#) values are comparable under these conditions. The pH-dependent increase in K(2)(#) with a pK(a) of 6.5 is attributed to E71 which participates in the binding of the second calcium which in turn influences the enzyme binding to phosphatidylcholine interface. Most mutants exhibited kinetic and spectroscopic behavior that is comparable to that of native PLA2 and DeltaPLA2 with a deleted 62-66 loop. However, the DeltaY52L substitution mutant cannot undergo the calcium-, pH-, or interface-dependent changes. We suggest that the Y52L substitution impairs the R to T transition and also hinders the approach of the Michaelis complex to the transition state. This allosteric change may be mediated by the structural motifs that connect the D48 D99 catalytic diad, the substrate-binding slot, and the residues of the i-face. Our interpretation is that the 57-72 loop and the H(48)DNCY(52) segment of PLA2 are involved in transmitting the effect of the cooperative amphiphile binding to the i-face as a structural change in the active site. PMID- 19301849 TI - Generation of nonagglomerated airborne bacteriophage particles using an electrospray technique. AB - Biological electrospray techniques are rapidly becoming a promising means for controlling living organisms in applications ranging from mass spectrometry to developmental biology. We investigated the generation characteristics of airborne MS2 bacteriophage particles <30 nm in size, using an electrospray technique. A suspension containing bacteriophage MS2 was sprayed in cone-jet mode using a specially designed electrospray system with a point-to-orifice-plate configuration mentioned in previous studies based on a charge reduced electrospray size spectrometry. The highly charged droplets were discharged rapidly into a radioactive neutralizer of Po(210). The electrosprayed airborne MS2 particles (23.8 +/- 0.49 nm GMD) maintained their monodisperse size distribution with good stability and uniformity for >1 h. Compared with the generation characteristics observed using the previous nebulization process (51.5 +/- 0.86 nm GMD), this electrospray technique produced nonagglomerated particles, resulting in a narrow size range of generated particles. The total MS2 particle number concentration and GMD increased with changes in the suspension flow rate from 5 to 25 microL/h. As the applied voltage increased in cone-jet mode, the GMD and culturable bacteriophage concentration decreased slightly. Our investigation shows that the electrospray process, driven by high-intensity electric fields, can be used for nanometer-sized living organisms. PMID- 19301850 TI - Efficient N-arylation/dealkylation of electron deficient heteroaryl chlorides and bicyclic tertiary amines under microwave irradiation. AB - A highly efficient procedure was developed for the microwave-assisted synthesis of N-heteroaryl-4-(2-chloroethyl)piperazines and N-heteroaryl-4-(2 chloroethyl)piperidines. Microwave irradiation of electron deficient heteroaryl chlorides with 1,4-diazabicyclo[2.2.2]octane (DABCO) at 160 degrees C for 15 min led to N-heteroaryl-4-(2-chloroethyl)piperazines in good to excellent yields. In a similar manner, microwave irradiation of electron deficient heteroaryl chlorides with quinuclidine at 180 degrees C for 15 min provided N-heteroaryl-4 (2-chloroethyl)piperidines in good to excellent yields. Extension of the method was demonstrated by the development of a one-pot, two-step microwave-assisted protocol for the synthesis of 4-(2-acetoxyethyl)-substituted N heteroarylpiperazines and N-heteroarylpiperidines to demonstrate the production of a small library in a parallel fashion. PMID- 19301851 TI - Controlled synthesis of polylactides using biogenic creatinine carboxylate initiators. AB - Two biogenetic guanidine carboxylates, i.e., creatinine acetate (CRA) and creatinine glycolate (CRG) based on creatinine (CR) were synthesized and structurally characterized by (1)H NMR, (13)C NMR and X-ray diffraction (XRD) analysis of the single crystals. The characterization demonstrated that CRA and CRG are CR-guanidinium carboxylates in nature, and the carboxylates are associated with the guanidinium via hydrogen bonds. CRA and CRG were successfully used as single-component initiators for ring-opening polymerizations (ROPs) of l lactide (LLA), dl-lactide (DLLA) producing polymers with controlled molecular weights and narrow polydispersities (PDI = 1.16-1.26). The investigation on the kinetics of the ROPs indicated the typical characteristics of living polymerization as evidenced by the linear relationship of ln [M](0)/[M](t) versus time and M(n) (number-average molecular weight of formed polymer) versus monomer conversion. A study of the terminator-adding effect on the growing polymers revealed that the polymerization follows the coordination propagation mode. The structures of the living oligomers and product polymers were characterized with (1)H NMR spectroscopy. The mechanism of the polymerization was proposed on the basis of the experimental investigation. PMID- 19301852 TI - Fluorous parallel synthesis of a piperazinedione-fused tricyclic compound library. AB - A fluorous-linker-assisted solution-phase protocol has been developed and applied to parallel synthesis of a piperazinedione-fused tricyclic compound library. The one-pot [3 + 2] cycloaddition of fluorous amino esters, aldehydes, and maleimides afforded bicyclic proline derivatives. The intermediates were subjected to N acylation with chloroacetyl chloride, followed by displacement reactions with amines. Linker cleavage with concomitant lactamization yielded the final products. Microwave heating was employed to facilitate several reaction steps and fluorous solid phase extraction (F-SPE) was employed to purify the intermediates. During the method development, a small library containing sixteen analogs was prepared. The optimized conditions were applied to the synthesis of a production library containing ninety analogs. PMID- 19301853 TI - Solution-phase synthesis of a library of carbapeptide analogues based on glycosylamino acid scaffolds and their in silico screening and antimicrobial evaluation. AB - A well-organized and efficient approach toward the solution phase synthesis of a library of carbapeptide analogues based on glycosyl amino ester scaffold is described. The reported synthetic route involves a five step preparation of heptofuranuronamides 6a-h and octopyranuronamide 7e from glycosyl amino esters 1 and 7, respectively. Coupling of glycosyl amino esters 1 or 7 with three different N-Fmoc protected amino acids afford the N-Fmoc protected intermediates 2a-c and 7a. Deprotection of Fmoc group in 2a-c and 7a with piperidine gave respective compounds 3a-c and 7b with free amine. Subsequent coupling of 3a-c and 7b with different aromatic acids furnishes respective heptofuranuronates 4a-h and octopyranuronate 7c in good yields. The latter, on ester hydrolysis by LiOH gave the corresponding glycopeptide analogues 5a-h and 7d with terminal carboxyl group. The carboxyl group in these compounds was amidated with oxalyl chloride/ NH(4)OH to afford heptofuranuronamides 6a-h and octopyranuronamides 7e. In vitro screening of all compounds displayed moderate antifungal, antitubercular, and general antibacterial activities. Reverse docking calculations involving over 841 protein drug targets have identified two potential targets for these compounds. These results will form the basis for synthesizing second-generation antimicrobial compounds. PMID- 19301854 TI - What singles out the FeO2+ moiety? A density-functional theory study of the methane-to-methanol reaction catalyzed by the first row transition-metal oxide dications MO(H2O)(p)2+, M = V-Cu. AB - Metaloxo species are often postulated as key active species in oxidative catalysis. Among all, the quintet FeO2+ moiety is particularly widespread and active: aliphatic C-H bonds undergo hydroxylation easily through a H abstraction/O-rebound mechanism. The high electrophilicity of quintet FeO2+ originates from its electronic structure: a low lying vacant sigma* can accept electronic density from the aliphatic C-H bond. What singles out this quintet FeO2+? Its lowest vacant acceptor orbital energy? its shape (sigma* vs pi*)? or has its biological importance more simply arisen from the high iron abundance? To answer those questions, we have performed density-functional theory calculations to study systematically the methane-to-methanol reaction catalyzed by MO(H2O)(p)2+ complexes (M = V, Cr, Mn, Fe, Co, p = 5 and M = Ni, Cu, p = 4) in gas phase. We show here that the lower the MO2+ acceptor orbital lies in energy, the lower the H-abstraction barrier is in general. However, a sigma* acceptor orbital is much more efficient than a pi* acceptor orbital for a given energy. Finally, we found that indeed, the FeO2+ moiety is particularly efficient but also CoO2+ and MnO2+ could be good candidates to perform C-H hydroxylation. PMID- 19301856 TI - Identification of an overabundant cholesterol precursor in hepatitis B virus replicating cells by untargeted lipid metabolite profiling. AB - Viruses rely upon host lipid metabolic pathways for successful replication, and there is increasing interest in these pathways as novel therapeutic targets for antiviral drug discovery. Despite this, relatively little is known about the impact of viral infection on cellular lipid metabolism, and the specific lipid metabolites utilized by viruses have not yet been examined. We have applied liquid chromatography-mass spectroscopy (LC-MS) based untargeted metabolite profiling to identify lipid metabolites whose steady-state abundance is significantly altered by replication of hepatitis B virus (HBV), a major human pathogen. Untargeted metabolite profiling indicated that although major lipid classes were unaffected by HBV, an ion of 367 m/z was overabundant in HBV+ cells by 18-fold. As shown by ion fragmentation mass spectrometry and coinjection with standard, the identity of this ion is 7-dehydrocholesterol (7-DHC), an immediate dehydrogenated precursor to cholesterol. While cholesterol has previously been demonstrated to be essential in the replication of many viruses, this is the first to show that viral replication is associated with the selective accumulation of 7-DHC. Most virological studies to date have relied upon methods that deplete all sterols and preclude the observation of any selectivity in sterol utilization by viral pathogens. Our study suggests that HBV may selectively utilize 7-DHC versus other sterols and prompts experiments investigating the functional significance of this enrichment and the elucidation of the mechanism by which it is achieved. The results also highlight the value of untargeted metabolite profiling as a method for identifying critical metabolites for viral infection. PMID- 19301857 TI - Ionic screening of charged-impurity scattering in graphene. AB - We have studied the ionic screening effect on the charge transport properties of graphene field effect transistors. By increasing the ionic concentration, we found dramatic increases in the carrier mobilities and systematic changes in the position and magnitude of minimum conductivity, as well as the width of minimum conductivity plateau, which supports the theory of long-range Coulomb scattering. We also observed clear conductivity saturation and systematic crossover from the linear to constant conductivity regimes. PMID- 19301858 TI - High-quality Graphenes via a facile quenching method for field-effect transistors. AB - Single- and few-layer graphene sheets with sizes up to 0.1 mm were fabricated by simply quenching hot graphite in an ammonium hydrogen carbonate aqueous solution. The identity and thickness of graphene sheets were characterized with transmission electron microscopy, atomic force microscopy, and Raman spectroscopy. In addition to its simplicity and scalability, the present synthesis can produce graphene sheets with excellent qualities in terms of sizes, purity, and crystal quality. The as-produced graphene sheets can be easily transferred to solid substrates for further processing. Field-effect transistors based on individual graphenes were fabricated and shown to have high ambipolar carrier mobilities. PMID- 19301859 TI - Magnetic tweezers measurement of single molecule torque. AB - Torsional stress in linear biopolymers such as DNA and chromatin has important consequences for nanoscale biological processes. We have developed a new method to directly measure torque on single molecules. Using a cylindrical magnet, we manipulate a novel probe consisting of a nanorod with a 0.1 microm ferromagnetic segment coupled to a magnetic bead. We achieve controlled introduction of turns into the molecule and precise measurement of torque and molecule extension as a function of the number of turns at low pulling force. We show torque measurement of single DNA molecules and demonstrate for the first time measurements of single chromatin fibers. PMID- 19301860 TI - Plasmon-enhanced single photon emission from a nanoassembled metal-diamond hybrid structure at room temperature. AB - In this Letter we present the controlled coupling of a single nitrogen vacancy center to a plasmonic structure. With the help of an atomic force microscope, a single nanodiamond containing a single nitrogen vacancy center and two gold nanospheres are assembled step-by-step. We show that both the excitation rate and the radiative decay rate of the color center are enhanced by about 1 order of magnitude, while the single photon character of the emission is maintained. Hot spots between diamond and gold nanoparticles provide an efficient near-field coupling, despite the mismatch in size and shape. Our approach provides hybrid systems as important building blocks for novel nanophotonic light sources in advanced plasmonic devices stable even at room temperature. PMID- 19301861 TI - Enhanced harvesting of red photons in nanowire solar cells: evidence of resonance energy transfer. AB - Modern excitonic solar cells efficiently harvest photons in the 350-650 nm spectral range; however, device efficiencies are typically limited by poor quantum yields for red and near-infrared photons. Using Forster-type resonance energy transfer from zinc phthalocyanine donor molecules to ruthenium polypyridine complex acceptors, we demonstrate a four-fold increase in quantum yields for red photons in dye-sensitized nanowire array solar cells. The dissolved donor and surface anchored acceptor molecules are not tethered to each other, through either a direct chemical bond or a covalent linker layer. The spatial confinement of the electrolyte imposed by the wire-to-wire spacing of the close-packed nanowire array architecture ensures that the distances between a significant fraction of donors and acceptors are within a Forster radius. The critical distance for energy transfer from an isolated donor chromophore to a self-assembled monolayer of acceptors on a plane follows the inverse fourth power instead of the inverse sixth power relation. Consequently, we observe near quantitative energy transfer efficiencies in our devices. Our results represent a new design paradigm in excitonic solar cells and show it is possible to more closely match the spectral response of the device to the AM 1.5 solar spectrum through use of electronic energy transfer. PMID- 19301863 TI - Rh(I)-catalyzed CO gas-free carbonylative cyclization reactions of alkynes with 2 bromophenylboronic acids using formaldehyde. AB - The rhodium(I)-catalyzed reaction of alkynes with 2-bromophenylboronic acids in the presence of paraformaldehyde resulted in a CO gas-free carbonylative cyclization, yielding indenone derivatives. [RhCl(BINAP)](2) and [RhCl(cod)](2) were responsible for the decarbonylation of formaldehyde and the subsequent carbonylation of alkynes with 2-haloboronic acids, respectively, leading to efficient whole carbonylation. Sterically bulky and electron-withdrawing groups on unsymmetrically substituted alkynes favored the alpha-position of indenones. PMID- 19301864 TI - Stereoelectronic versus steric tuning in the prins cyclization reaction: synthesis of 2,6-trans pyranyl motifs. AB - The use of carboalkoxyl allenic alcohol for the efficient synthesis of pyranyl motifs via Prins cyclization is described. This method provides easy access to 2,6-trans dihydropyrans in good yield and high diastereoselectivity. PMID- 19301865 TI - Ring strain energy in the cyclooctyl system. The effect of strain energy on [3 + 2] cycloaddition reactions with azides. AB - Ring strain energies (SEs) and enthalpies of hydrogenation (DeltaH(hyd)) of a series of E- and Z-alkenes, cyclic alkynes and allenes (C(5)-C(9)) are computed at the G3 level of theory. The SE for cycloheptyne, cyclohexyne, and cyclopentyne are calculated to be 25.4, 40.1, and 48.4 kcal/mol, respectively. The SE for E cycloheptene and E-cyclohexene are calculated to be 25.2 and 49.3 kcal/mol (G3). The SE of cyclooctyne is 2.0 kcal/mol greater than that of E-cyclooctene (17.9 kcal/mol) but only 7.7 kcal/mol greater than that of cyclooctane. The SE of 3,3 difluorocyclooctyne (DIFO) is predicted to be slightly reduced (DeltaSE = 2.6 kcal/mol) relative to the parent cyclooctyne to 17.3 kcal/mol. The SE and DeltaH(hyd) are correlated with activation barriers for the [3 + 2] cycloaddition of a series of azides to E- and Z-cycloalkenes and alkynes at the G3 level of theory. The energy barrier for the cycloaddition of methyl azide to cyclooctyne is 9.2 kcal/mol lower than addition to 4-octyne and 3.1 kcal/mol lower for reaction with E-cyclooctene. The activation energies for [3 + 2] cycloaddition of benzyl azide and acetamido azide ((2)HN(C=O)CH(2)-N(3)) to DIFO are 2.3 and 5.3 kcal/mol lower in energy than cycloaddition to cyclooctyne [B3LYP/6 311+G(3df,2p)]. PMID- 19301866 TI - Translation initiation with initiator tRNA charged with exotic peptides. AB - In the prokaryotic translation system formylmethionyl-tRNA(fMet)(CAU) acts as an exclusive initiator to yield peptides bearing formylmethionine at the N-terminus. We herein report a new methodology to initiate the translation reaction with peptidyl-tRNA(fMet)(CAU) in which the peptidyl group consists of unusual building blocks, such as D-amino acids, beta-amino acids, and N-methyl amino acids, and express peptides containing a stretch of exotic peptides at the N-terminus. Synthesis of the initiator peptidyl-tRNAs was facilitated by flexizyme, a flexible tRNA aminoacylation ribozyme, and the start codon was reprogrammed by withdrawing methionine from a reconstituted E. coli cell-free translation system to reassign the initiator from formylmethionine to exotic peptides. This represents the first demonstration for initiating the ribosomal peptide synthesis with peptides, which offers us a new tool for the synthesis of a wide variety of unusual peptides and also the mechanistic studies on the initiation and/or elongation events in translation. PMID- 19301867 TI - Shuttling germanium atoms into branched polysilanes. AB - Lewis acid-catalyzed rearrangement reactions of trimethylgermylated small permethylated oligosilanes were found to show a strong tendency of the germanium atoms to move to the more highly silylated positions of the molecule. This makes possible the construction of molecules with high germanium content in the core parts of the molecules by the following repeated sequence: introduction of trimethylgermyl group followed by rearrangement. Quantum-mechanical computations at the B3LYP/6-311+G(d,p) level suggest that the exergonic rearrangement proceeds in a reaction cascade via germylium and silylium ions, which transform by 1,2 silyl and -methyl shifts. PMID- 19301868 TI - On the mechanism of Trolox as antiblinking and antibleaching reagent. AB - Recent advances in photobleaching and blinking prevention have aided the advancement of single-molecule and super-resolution fluorescence microscopy. However, a common mechanism of the action of antifading agents such as Trolox is still missing. In this communication we present evidence that Trolox acts in accordance with a mechanism that involves triplet quenching through electron transfer and subsequent recovery of the resulting radical ion by the complementary redox reaction. The required oxidant for this unifying mechanism based on a reducing and oxidizing system (ROXS) is formed via (photo-) reaction with molecular oxygen. We present evidence that this oxidized form is a quinone derivative of Trolox with strong oxidizing properties. These findings shed light on many contradicting results regarding the action of antifading agents and might lead to a common mechanistic understanding of photobleaching and its prevention. Finally, a recipe on the proper use of Trolox as an antifading agent is provided. PMID- 19301870 TI - Oxidant-controlled heck-type C-glycosylation of glycals with arylboronic acids: stereoselective synthesis of aryl 2-deoxy-C-glycosides. AB - Oxidative Heck-type C-glycosylations of glycals with various arylboronic acids using Pd(OAc)(2) as catalyst in the presence of oxidant were developed. The corresponding ketone, enol ether, and enone types of C-glycosides were predictably obtained with benzoquinone (BQ), Cu(OAc)(2)/O(2), and 2,3-dichloro 5,6-dicyano-1,4-benzoquinone (DDQ) as oxidants, respectively. This method provides a simple, mild, and stereoselective synthesis of aryl 2-deoxy-C glycosides. PMID- 19301871 TI - Reaction mechanism of glutamate carboxypeptidase II revealed by mutagenesis, X ray crystallography, and computational methods. AB - Glutamate carboxypeptidase II (GCPII, EC 3.4.17.21) is a zinc-dependent exopeptidase and an important therapeutic target for neurodegeneration and prostate cancer. The hydrolysis of N-acetyl-l-aspartyl-l-glutamate (N-Ac-Asp Glu), the natural dipeptidic substrate of the GCPII, is intimately involved in cellular signaling within the mammalian nervous system, but the exact mechanism of this reaction has not yet been determined. To investigate peptide hydrolysis by GCPII in detail, we constructed a mutant of human GCPII [GCPII(E424A)], in which Glu424, a putative proton shuttle residue, is substituted with alanine. Kinetic analysis of GCPII(E424A) using N-Ac-Asp-Glu as substrate revealed a complete loss of catalytic activity, suggesting the direct involvement of Glu424 in peptide hydrolysis. Additionally, we determined the crystal structure of GCPII(E424A) in complex with N-Ac-Asp-Glu at 1.70 A resolution. The presence of the intact substrate in the GCPII(E424A) binding cavity substantiates our kinetic data and allows a detailed analysis of GCPII/N-Ac-Asp-Glu interactions. The experimental data are complemented by the combined quantum mechanics/molecular mechanics calculations (QM/MM) which enabled us to characterize the transition states, including the associated reaction barriers, and provided detailed information concerning the GCPII reaction mechanism. The best estimate of the reaction barrier was calculated to be DeltaG(++) approximately 22(+/-5) kcal x mol(-1), which is in a good agreement with the experimentally observed reaction rate constant (k(cat) approximately 1 s(-1)). Combined together, our results provide a detailed and consistent picture of the reaction mechanism of this highly interesting enzyme at the atomic level. PMID- 19301872 TI - Supramolecular chemistry of metalloporphyrins. PMID- 19301874 TI - Development and validation of a novel protein-ligand fingerprint to mine chemogenomic space: application to G protein-coupled receptors and their ligands. AB - The present study introduces a novel low-dimensionality fingerprint encoding both ligand and target properties which is suitable to mine protein-ligand chemogenomic space. Whereas ligand properties have been represented by standard descriptors, protein cavities are encoded by a fixed length bit string describing pharmacophoric properties of a definite number of binding site residues. In order to simplify the cavity fingerprint, the concept was applied here to a unique family of targets (G protein-coupled receptors) with a homogeneous cavity description. Particular attention was given to set up data sets of really diverse protein-ligand pairs covering as exhaustively as possible both ligand and target spaces. Several machine learning classification algorithms were trained on two sets of roughly 200000 receptor-ligand fingerprints with a different definition of inactive decoys. Cross-validated models show excellent precision (>0.9) in distinguishing true from false pairs with a particular preference for support vector machine classifiers. When applied to two external test sets of GPCR ligands, the most predictive models were not those performing the best in the previous cross-validation. The ability to recover true GPCR ligands (ligand prediction mode) or true GPCRs (receptor prediction mode) depends on multiple parameters: the molecular complexity of the ligands, the chemical space from which ligand decoys are selected to generate false protein-ligand pairs, and the target space under consideration. In most cases, predicting ligands is easier than predicting receptors. Although receptor profiling is possible, it probably requires a more detailed description of the ligand-binding site. Noteworthy, protein-ligand fingerprints outperform the corresponding ligand fingerprints in mining the GPCR-ligand space. Since they can be applied to a much larger number of receptors than ligand-based fingerprints, protein-ligand fingerprints represent a novel and promising way to directly screen protein-ligand pairs in chemogenomic applications. PMID- 19301875 TI - Combinatorial library of primaryalkylammonium dicarboxylate gelators: a supramolecular synthon approach. AB - Following the supramolecular synthon approach, a combinatorial library comprising 35 organic salts derived from 7 dicarboxylic acids (malonic-, succinic-, adipic-, L-tartaric-, maleic-, phthalic-, and isophthalicacid) and 5 primaryalkyl amines Me-(CH2)n-NH2 (n = 11-15) was prepared and scanned for gelation. About 66% of the salts in the combinatorial library were found to show moderate to good gelling ability in various polar and nonpolar solvents including commercial fuels such as petrol. The majority of the salts having a rigid, unsaturated anionic backbone (maleate, phthalate, and isophthalate) did not show gelation; only the corresponding hexadecylammonium salts showed gelation. Some of the representative gels were characterized by rheology, small-angle neutron scattering (SANS), optical microscopy (OM), and scanning electron microscopy (SEM). Single-crystal structures of two gelator and two nongelator salts were also discussed in the context of supramolecular synthon and structure-property correlation. PMID- 19301876 TI - Striking correlation between the unusual trigonal crystal packing and the ability to self-assemble into nanofibers of 2,3-di-n-alkyloxyanthracenes. AB - The space group of the crystals of derivatives of 2,3-dialkoxyanthracenes is monoclinic P2(1)/a (herringbone structure) with the linear ethyl or propyl chains but abruptly changes to the trigonal P3 or R3 space group for butyl to heptyl chains. Strikingly, this switch is correlated with the capacity of these compounds to self-assemble into nanofibers and organogels. Besides, compounds with a chain length exceeding seven carbon atoms could not be crystallized in accordance with the analysis of the projected crystal structure but are nevertheless excellent organogelators. The study of this series of compounds suggests a tight link between the molecular structure of the crystals and that of the organogels. PMID- 19301877 TI - Nanophase separation in polystyrene-polyfluorene block copolymers thin films prepared through the breath figure procedure. AB - The amphiphilic block copolymer formed by a hydrophobic body of polystyrene and a hydrophilic head of poly[9,9-di(2-(2-tetrahydropyranyl-oxy)hexyl)fluorene-alt-9,9 dioctylfluorene] was synthesized, and its solution was used to create thin films with ordered pattern of holes, by means of the breath figure technique. These porous films, after a thermal treatment, were found to show ordered aggregates of the pi-conjugated blocks in the place of the cavities. This is probably due to a preorganization of the two different blocks of the copolymer occurring during the breath figure formation, which is driven by the condensation of water microdroplets on the polymer solution, and to a following phase segregation occurring during the thermal annealing. This approach is a promising tool to be employed for the organization of organic materials at the micro and nanoscale. PMID- 19301878 TI - Molecular recognition by contact angle: proof of concept with DNA hybridization. AB - A molecular recognition reaction supported by a solid-phase assay drives a specific change in the solid-solution interfacial tension. This prompts contact angle (CA) analysis, being a straightforward route to evaluate this property, to play the unedited role of label-free probe of the reaction. The concept is proven by the successful recognition of DNA hybridization and is further supported by the agreement between the results and the underpinning thermodynamics. PMID- 19301879 TI - Reduced sterol-phospholipid recognition in curved fluid bilayers. AB - Nearest-neighbor recognition experiments have been carried out in fluid liposomal membranes made from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol using exchangeable dimers derived from 1,2-dipalmitoyl-sn-glycero-3 phosphoethanolamine and cholesterol. In cholesterol-rich bilayers, the association between these two exchangeable lipids was reduced as the curvature of the membrane increases; that is, when the diameter of the liposomes was below ca. 200 nm. In sharp contrast, the mixing of these exchangeable lipids was close to random in the cholesterol-poor membranes, regardless of their curvature. The biological implications of these findings are briefly discussed. PMID- 19301880 TI - Chiral assembly from achiral rod-coil molecules triggered by compression at the air-water interface. AB - A series of achiral rod-coil molecules consisting of an oligo(p-phenylene) conjugated rod with a poly(ethylene oxide) or poly(propylene oxide) coil laterally attached through an imidazole linkage were synthesized, and their interfacial behaviors were investigated. Compounds 1a and 1b have a similar surface behavior with a shoulder indicating a transformation of rod segments from flat-on to vertical orientation. Interestingly, CD and AFM suggest chiral films are formed when the LB films were deposited after the shoulder. Although compound 1c shows different surface behavior because the PEO chains dissolved into water upon compression, macroscopic chirality was also detected. In contrast, compound 2, based on a reduced conformational rigidity of the aromatic rod segment, does not form chiral assemblies. Combining the data collected, the cooperative interaction of the hydrogen bond between the molecules and pi-pi stacking as well as the steric constraint between the aromatic rod segments are responsible for the macroscopic chirality. Such kind of stacking can be realized through a molecular design and a lateral compression. A deep insight into the relationship between the molecular structure and the chirality was gained. PMID- 19301873 TI - Functional nucleic acid sensors. PMID- 19301881 TI - Re-entrant phase behavior of a concentrated anionic surfactant system with strongly binding counterions. AB - The phase behavior of the anionic surfactant sodium dodecyl sulfate (SDS) in the presence of the strongly binding counterion p-toluidine hydrochloride (PTHC) has been examined using small-angle X-ray diffraction and polarizing microscopy. A hexagonal-to-lamellar transition on varying the PTHC to SDS molar ratio (alpha) occurs through a nematic phase of rodlike micelles (Nc) --> isotropic (I) --> nematic of disklike micelles (N(D)) at a fixed surfactant concentration (phi). The lamellar phase is found to coexist with an isotropic phase (I') over a large region of the phase diagram. Deuterium nuclear magnetic resonance investigations of the phase behavior at phi = 0.4 confirm the transition from N(C) to N(D) on varying alpha. The viscoelastic and flow behaviors of the different phases were examined. A decrease in the steady shear viscosity across the different phases with increasing alpha suggests a decrease in the aspect ratio of the micellar aggregates. From the transient shear stress response of the N() and N(D) nematic phases in step shear experiments, they were characterized to be tumbling and flow aligning, respectively. Our studies reveal that by tuning the morphology of the surfactant micelles strongly binding counterions modify the phase behavior and rheological properties of concentrated surfactant solutions. PMID- 19301882 TI - A simple synthesis of nitriles from aldoximes. AB - Easily synthesized aldoximes have been converted to the corresponding nitriles under very mild conditions by a simple reaction with 1H-benzotriazol-1 yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP) and DBU in CH(2)Cl(2), THF, or DMF. As an alternative reagent that eliminates the formation of hexamethylphosphoramide as a byproduct, use of 1H-benzotriazol-1-yl-4 methylbenzenesulfonate (Bt-OTs) and DBU was investigated. Reactions with this reagent also proceeded smoothly and in good yields, although in one case N sulfonylation was observed. An attempt to gain mechanistic insight into the BOP mediated reaction has been made using (31)P{(1)H} NMR. However, no phosphorus bearing intermediate could be readily observed. Finally, the method has been applied to the synthesis of an antiviral 4'-cyano adenosine analogue from a commercial precursor using a single saccharide protecting group. PMID- 19301883 TI - Application of beta-oxodithioesters in domino and multicomponent reactions: facile route to dihydropyrimidines and coumarins. AB - A facile route to hitherto unknown 5-methylmercaptothiocarbonyl-4-aryl-3,4 dihydropyrimidin-2(1H)-ones and substituted 2H-chromene-2-thiones has been developed. SnCl(2)-catalyzed cyclocondensation of beta-oxodithioesters with a variety of readily accessible aldehydes and urea affords the dihydropyrimidinones. The methodology involves the three-component Biginelli reaction. On the other hand, substituted salicylaldehyde and beta-oxodithioesters reacted under the same condition to afford the substituted 2H-chromene-2-thiones in high yields. PMID- 19301884 TI - Methanesulfonamide: a cosolvent and a general acid catalyst in sharpless asymmetric dihydroxylations. AB - To obtain information about the effect that methanesulfonamide has in the hydrolysis step in Sharpless asymmetric dihydroxylation, a series of aliphatic and conjugated aromatic olefins were dihydroxylated with and without methanesulfonamide. The hypothesis in this study was that methanesulfonamide is a cosolvent that aids in the transfer of the hydroxide ions from the water phase to the organic phase. A plot of t90% versus the computational partition coefficient clog P of the intermediate osmate ester of nonterminal aliphatic olefins revealed that the polarity of the intermediate osmate ester has a significant effect on the reaction time and methanesulfonamide effect. The more polar the intermediate osmate ester, the faster is the reaction without methanesulfonamide and the smaller the accelerating methanesulfonamide effect. Methanesulfonamide had no accelerating effect in the asymmetric dihydroxylation of short chain terminal aliphatic olefins as a result of easier accessibility of terminal osmate ester groups to the water phase. A cosolvent hypothesis was found not to be valid in asymmetric dihydroxylations of conjugated aromatic olefins. In the reaction conditions used in Sharpless asymmetric dihydroxylation, weakly acidic methanesulfonamide was found to be a general acid catalyst that protonates the intermediate osmate esters of conjugated aromatic olefins in the hydrolysis step. PMID- 19301885 TI - Rational design of improved aziridine-based inhibitors of cysteine proteases. AB - Quantum chemical computations on appropriate model systems are used for a rational design of aziridine-based inhibitors of cysteine proteases. They predict that already inductive electron-withdrawing substituents at the aziridine nitrogen strongly accelerate the alkylation step of the inhibition process in neutral and alkaline media, but also for more acidic environments improvements are predicted. With this we generalize previous findings that found similar effects for N-formylated compounds. Furthermore, the new substituents possess the additional advantage that they do not open up reaction pathways other than the nucleophilic ring opening. To verify the hypotheses selected compounds were synthesized and tested. These tests approved the predictions and showed that the corresponding derivatives of aziridine-2,3-dicarboxylate are potent irreversible inhibitors of cysteine proteases. On the basis of measured inhibition data the new inhibitors offer an up to 2,300-fold increase in inhibition potency compared to the unsubstituted inhibitor. Additionally, the kinetics of a selected reaction with 4-methoxy thiophenolate as model thiol were measured in solution to ascertain that the inhibition mechanism is the irreversible alkylation of the cysteine residue of the protease's active site under ring opening of the new inhibitors. PMID- 19301886 TI - Study of ion transport in lithium perchlorate-succinonitrile plastic crystalline electrolyte via ionic conductivity and in situ cryo-crystallography. AB - Ion transport mechanism in lithium perchlorate (LiClO(4))-succinonitrile (SN), a prototype of plastic crystalline soft matter electrolyte is discussed in the context of solvent configurational isomerism and ion solvation. Contributions of both solvent configurational isomerism and ion solvation are reflected in the activation energy for ion conduction in 0-1 M LiClO(4)-SN samples. Activation energy due to solvent configurational changes, that is, trans-gauche isomerism is observed to be a function of salt content and decreases in presence of salt (except at high salt concentrations, e.g. 1 M LiClO(4)-SN). The remnant contribution to activation energy is attributed to ion-association. The X-ray diffraction of single crystals obtained using in situ cryo-crystallography confirms directly the observations of the ionic conductivity measurements. Fourier transform infrared spectroscopy and NMR line width measurements provide additional support to our proposition of ion transport in the prototype plastic crystalline electrolyte. PMID- 19301887 TI - Supramolecular ring banded prototype liquid crystalline oligo(phenylenevinylene). AB - We report a new ring banded supramolecular structure in thermotropic liquid crystalline oligo(phenylenevinylene) (OPV) via a melt crystallization process. A series of structurally different OPV molecules were synthesized using tricyclodecanemethanol (TCD) as a bulky pendant unit to trace ring banded morphology. Among all, an OPV molecule with rigid bis-TCD units in the central core and flexible dodecyl chains at the outer phenyl rings (BTCD-BDD-OPV) was found to show ring banded morphologies, which is a first of its kind in pi conjugated materials. BTCD-BDD-OPV experiences strong aromatic pi-pi interactions in both film and liquid crystalline (LC) frozen stage. The pi-induced aggregation leads to lamellar self-assembly of OPV-mesogens that subsequently undergo helical crystal growth, thereby producing dark and bright ring banded patterns. Variable temperature X-ray diffraction analysis revealed the existence of three peaks at 27.07, 13.97, and 8.90 A corresponding to 001, 002, and 003 fundamental layers, respectively, thus confirming the lamellar self-assembly of OPV-mesogens. Electron microscopic (SEM and TEM) analysis of the LC frozen sample showed images confirming helical microcrystalline assembly and providing direct evidence for the self-organization mechanism. Detailed photophysical experiments such as excitation, emission, and time-resolved fluorescence decay studies indicated that BTCD-BDD-OPV has very strong pi-pi interaction in both film and LC frozen stage, which was found to be main driving force for the formation of supra-ring structure. Upon illumination with light, the OPV chromophores in the LC phase were excited and the color of the samples turned into luminescent green ring bands. PMID- 19301888 TI - Miscibility between differently shaped mesogens: structural and morphological study of a phthalocyanine-perylene binary system. AB - The thermotropic, structural, and morphological properties of blends of a disk like liquid crystalline phthalocyanine derivative and a lath-shaped perylenetetracarboxidiimide mesogen derivative have been studied by combining differential scanning calorimetry, thermal polarized optical microscopy, X-ray diffraction, solid-state nuclear magnetic resonance, and atomic force microscopy. The two compounds are fully miscible for blends containing at least 60 mol % of the disk-like molecule. In such composition range, the homogeneous blends form a columnar hexagonal (Col(h)) mesophase for which the thermal stability is enhanced compared to that of the corresponding mesophase of the pure phthalocyanine. The miscible blends self-align homeotropically between two glass slides. For blends containing between 55 and 40 mol % of the disk-shaped molecule, the two components are fully miscible at high temperature but the perylene derivative forms a separate crystalline phase when the temperature is decreased. Phase separation is systematically observed in blends containing less than 40 mol % of the discotic molecule. In this case, the resulting Col(h) mesophase is less stabilized compared to the blends containing a larger amount of the phthalocyanine derivative. These phase-separated blends do not show any homeotropic alignment. AFM investigations confirm the formation of a single columnar morphology in the phthalocyanine-rich blends, consistent with the full miscibility between the two compounds. Solid-state NMR measurements on the mixed phase show the influence of the presence of the perylene molecules on the molecular dynamics of the molecules; remarkably, the presence of the host molecules improves the local order parameter in the phthalocyanine columnar phase. PMID- 19301889 TI - Polyaniline/Fe3O4 nanoparticle composite: synthesis and reaction mechanism. AB - Polyaniline/Fe(3)O(4) nanoparticle composite was prepared by polymerizing aniline in the presence of Fe(3)O(4) nanoparticles upon the use of H(2)O(2) as oxidant. The polymerization was monitored by ultraviolet-visible absorption spectroscopy. The microstructure of the resultant composite was characterized by transmission electron microscopy. The molecular structure of the resultant polyaniline in the composite was investigated by both Fourier transform infrared spectroscopy and X ray photoelectron spectroscopy, while the magnetic property of the composite was characterized by vibrating sample magnetometer. Furthermore, the microwave absorption property of the resultant composite was measured in a frequency range of 2-18 GHz. Systematic investigations revealed that carboxylic acid in the buffer presented a determined role in the polymerization of aniline. To discover the role of carboxylic acid in the polymerization of aniline, more control experiments were designed and carried out by theoretical calculation in combination with electron spin resonance measurements. It was for the first time found out that carboxylic acid such as acetic acid and succinic acid can not only catalyze the polymerization of aniline but also facilitate the generation of hydroxyl radical via the decomposition of H(2)O(2). PMID- 19301890 TI - Theoretical and experimental studies of the reactions between hyperthermal O(3P) and graphite: graphene-based direct dynamics and beam-surface scattering approaches. AB - Beam-surface scattering experiments and theoretical direct dynamics based on density functional theory calculations are used to investigate hyperthermal collisions between O((3)P) and highly oriented pyrolytic graphite (HOPG). The simulations suggest that the HOPG surface becomes functionalized with epoxide groups. Intersystem crossing (ISC) between the lowest-energy triplet and singlet potential-energy surfaces is not necessary for this functionalization to occur. Both theory and experiment indicate that incoming O atoms can react at the surface to form O(2) by way of an Eley-Rideal mechanism. They also suggest that the collisions can result in the production of CO and CO(2) by way of both direct and complex reaction mechanisms. The direct dynamics simulations provide significant insight into the details of the complex reaction mechanisms. Semiquinones are present at defect sites and can form in functionalized pristine sheets, the latter resulting in the formation of a defect. Direct collision of an incoming O atom with a semiquinone or vibrational excitation caused by a nearby O atom collision can cause the release of the semiquinone CO, forming carbon monoxide. The CO may react with an oxygen atom on the surface to become CO(2) before receding from the surface. The simulations also illustrate how epoxide groups neighboring semiquinones catalyze the release of CO. Throughout, the experimental results are observed to be consistent with the theoretical calculations. PMID- 19301891 TI - Matrix photochemistry, photoelectron spectroscopy, solid-phase structure, and ring strain energy of beta-propiothiolactone. AB - The four-membered heterocyclic beta-propiothiolactone compound was isolated in a low-temperature inert Ar matrix, and the UV-visible (200 < or = lambda < or = 800 nm) induced photochemistry was studied. On the basis of the IR spectra, the formation of methylketene (CH(3)CHCO) was identified as the main channel of photodecomposition. The formation of ethene and thiirane, with the concomitant elimination of OCS and CO, respectively, was also observed as minor decomposition channels. The valence electronic structure was investigated by HeI photoelectron spectroscopy assisted by quantum chemical calculations at the OVGF/6-311++G(d,p) level of theory. The first three bands at 9.73, 9.87, and 12.06 eV are ascribed to the n''(S), n'(O), and pi''(CO) orbitals, respectively, denoting the importance of the -SC(O)- group in the outermost electronic properties. Additionally, the structure of a single crystal, grown in situ, was determined by X-ray diffraction analysis at low temperature. The crystalline solid [monoclinic system, P21/c, a = 8.1062(1) A, b = 10.3069(2) A, c = 10.2734(1) A, beta = 107.628(1) degrees, and Z = 8] consists of planar molecules arranged in layers. The skeletal parameters, especially the valence angles [angleC2-C1-S = 94.55(7) degrees, angleOC-C = 134.20(11) degrees, angleC-S-C = 77.27(5) degrees], differ from those typically found in acyclic thioester compounds, suggesting the presence of strong strain effects. The conventional ring strain energy was determined to be 16.4 kcal/mol at the G2MP2 level of calculation within the hyperhomodesmotic model. PMID- 19301892 TI - Charge effect on the diffusion coefficient and the bimolecular reaction rate of diiodide anion radical in room temperature ionic liquids. AB - The diffusion coefficients of diiodide anion radical, I(2)(-), in room temperature ionic liquids (RTILs) were determined by the transient grating (TG) method using the photochemical reaction of iodide. The diffusion coefficients we obtained were larger in RTILs than the theoretical predictions by the Stokes Einstein relation, whereas both values are similar in conventional solvents. By comparison with the diffusion coefficients of neutral molecules, it was suggested that the Coulomb interaction between I(2)(-) and constituent ions of RTILs strongly affects the diffusion coefficients. The bimolecular reaction rates between I(2)(-) were calculated by the Debye-Smoluchowski equation using the experimentally determined diffusion coefficients. These calculated reaction rate were much smaller than the experimentally determined rates (Takahashi, K.; et al. J. Phys. Chem. B 2007, 111, 4807), indicating the charge screening effect of RTILs. PMID- 19301893 TI - Effect of halogenation on the mechanism of the atmospheric reactions between methylperoxy radicals and NO. A computational study. AB - The mechanism of the reactions between the halogenated methylperoxy radicals, CHX(2)O(2) (X = F, Cl), and NO is investigated by using ab initio and density functional quantum mechanical methods. Comparison is made with the mechanism of the CH(3)O(2) + NO reaction. The most important energy minima in the potential energy surface are found to be the two conformers of the halogenated methyl peroxynitrite association adducts, CHX(2)OONOcp and CHX(2)OONOtp, and the halogenated methyl nitrates, CHX(2)ONO(2). The latter are suggested to be formed through the one-step isomerization of the peroxynitrite adduct and may lead upon decomposition to carbonylated species, CX(2)O + HONO and CHXO + XNO(2). The ambiguous issue of the unimolecular peroxynitrite to nitrate isomerization is reconsidered, and the possibility of a triplet transition state involvement in the ROONOtp <--> RONO(2) rearrangement is examined. The overall calculations and the detailed correlation with the methyl system show the significant effect of the halogenation on the lowering of the entrance potential energy well which corresponds to the formation of the peroxynitrites. The increased attractive character of the potential energy surface found upon halogenation combined with the increased exothermicity of the CHX(2)O(2) + NO --> CHX(2)O + NO(2) reaction are suggested to be the important factors contributing to the enhanced reactivity of the halogenated reactions relative to CH(3)O(2) + NO. The calculated heat of formation values indicate the large stabilization of the fluorinated derivatives. PMID- 19301894 TI - Effects of alkylation on deviations from Lennard-Jones collision rates for highly excited aromatic molecules: collisions of methylated pyridines with HOD. AB - Collision rates and energy transfer distributions are reported for HOD with highly vibrationally excited 2-methylpyridine (2-picoline, E = 38 310 cm(-1)) and 2,6-dimethylpyridine (2,6-lutidine, E = 38 700 cm(-1)). High resolution transient IR absorption is used measured to complete product state distributions of scattered HOD(000) molecules with E(rot) = 109 to 1331 cm(-1). Doppler-broadened line profiles characterize the depletion and appearance for HOD molecules due to collisions with hot donors and show that the product translational and rotational energy distributions are similar for both donors with DeltaE(rel) = 370 cm(-1) and DeltaE(rot) approximately 75 cm(-1). The energy transfer rate for picoline (E)/HOD is 2.5 times larger than the Lennard-Jones collision rate. The energy transfer rate for lutidine(E)/HOD is 3.2 times larger than the Lennard-Jones collision rate. Previous work ( Havey, Liu, Li, Elioff, and Mullin, J. Phys. Chem. A 2007, 111, 13321-9 ) reported similar energy transfer values for pyrazine/HOD collisions and an energy transfer rate that is 1.7 times the Lennard Jones collision rate. The observed collision rates are discussed in terms of hydrogen bonding interactions between HOD and the aromatic donor molecules. Energy gain profiles for HOD are compared with those for H(2)O. PMID- 19301895 TI - pH-controlled self-assembling of meso-tetrakis(4-sulfonatophenyl)porphyrin chitosan complexes. AB - Solid meso-tetrakis(4-sulfonatophenyl)porphyrin (TPPS(4))-chitosan supramolecular complexes were prepared by addition of porphyrin to an aqueous solution of chitosan at pH values. The precipitates obtained were assigned as 1 (pH 6.8) and 2 (pH 2.5) and characterized by spectroscopic, thermal, and microscopic methods. Spectroscopic investigation confirmed the presence of TPPS(4) and chitosan in both products and that the porphyrin is highly self-associated. H-type (stacked) of TPPS(4) aggregation was proposed for 1 and J-type (tilted) for 2. Thermal analysis revealed different pyrolysis routes of the complexes depending on their structural diversity. Light microscopic analysis indicated fibrous and lamellar microstructures, respectively, for 1 and 2. SEM and AFM analysis showed that both complexes consist of compact nanostructures; their size and interconnection is different for 1 and 2. Based on structural inferences, self-assembling hierarchy models were proposed for both of the TPPS(4)-chitosan supramolecular complexes. PMID- 19301896 TI - 2-DE and LC-MS/MS for a comparative proteomic analysis of BALf from subjects with different subsets of inflammatory myopathies. AB - The protein profiles of bronchoalveolar lavage fluid (BALf) of patients belonging to three selected subsets of Polymyositis/Dermatomyositis (PM/DM) have been compared by using a combination of 2-DE and MALDI-TOF/MS or LC-MS/MS. Our study examined the hypothesis that there were distinct differences in protein expression profiles that were related to the phenotype. From among the 323+/-51 protein spots that may represent the most highly expressed proteins in BALf of these patients, 24 unique spots were isolated and proteins identified. In particular, 9 spots were present in BALf of PM/DM patients only; 12 spots were exclusive of Overlap patients and 3 spots of AS patients. From among the proteins identified, a few were classified as cytoskeletal proteins, others were involved in oxidative stress and a number of proteins were associated with general metabolic activity or immunological response and inflammation. This is the first study in which evidence is provided that a number of different proteins are expressed in different subsets of PM/DM and supports our contention that the proteomic approach would be beneficial in discovering molecules which could represent possible prognostic factors of these rare pathologies. PMID- 19301897 TI - Design and structural analysis of novel pharmacophores for potent and selective peroxisome proliferator-activated receptor gamma agonists. AB - Utilizing medicinal chemistry design strategies such as benzo splitting and ring expansion, we converted PPARalpha/gamma dual agonist 1 to selective PPARgamma agonists 19 and 20. Compounds 19 and 20 were 2- to 4-fold better than rosiglitazone at PPARgamma receptor, with 80- to 100-fold PPARgamma selectivity over PPARalpha receptor. X-ray cocrystal studies in PPARgamma and modeling studies in PPARalpha give molecular insights for the improved PPARgamma potency and selectivity for 19 when compared to 1. PMID- 19301898 TI - Complementary three-dimensional quantitative structure-activity relationship modeling of binding affinity and functional potency: a study on alpha4beta2 nicotinic ligands. AB - Complementary 3D-QSAR modeling of binding affinity and functional potency is proposed as a tool to pinpoint the molecular features of the ligands, and the corresponding amino acids in the receptor, responsible for high affinity binding vs those driving agonist behavior and receptor activation. This approach proved successful on a series of nicotinic alpha(4)beta(2) ligands, whose partial/full agonist profile could be linked to the size of the scaffold as well as to the nature of the substituents. PMID- 19301899 TI - Gold-catalyzed homogeneous oxidative C-O bond formation: efficient synthesis of 1 benzoxyvinyl ketones. AB - A novel Au-catalyzed homogeneous oxidative C-O bond-forming reaction involving a Au(I)/Au(III) catalytic cycle is developed. Mechanistic studies reveal the involvement of a unique intramolecular carboxy migration. From readily available propargylic benzoates, this chemistry allows efficient access to captodative alkenes and dienones, demonstrating the synthetic potential of incorporating Au(I)/Au(III) catalytic cycles into contemporary Au chemistry. The unique reactivity and the mechanistic insights would help open a new research area in gold catalysis. PMID- 19301900 TI - Columnar packing motifs of functionalized perylene derivatives: local molecular order despite long-range disorder. AB - We elucidate local packing motifs and dynamical order parameters in a perylene tetracarboxydiimide derivative (C(8,7)-PDI), one of the most promising candidates for rationally designed, self-assembling, and self-healing molecular wires. Spectroscopic fingerprints obtained from solid-state NMR spectroscopy are interpreted by means of first-principles calculations and molecular dynamics simulations. The interplay of steric repulsion, H bonding, and pi-pi packing effects leads to a specific relative molecular pitch angle of approximately 35 +/ 10 degrees between successive molecules in the stack. Dynamical order parameters, determined from NMR sideband patterns as a measure of molecular motion, yield values of S approximately = 1.0 in the core of the columnar stack, corresponding to an almost frozen molecular dynamics at ambient temperature. This rigidity is compatible with characteristic intermolecular distances obtained from dipolar couplings between specific hydrogens via double-quantum NMR experiments and further supported by ab initio calculations. PMID- 19301901 TI - A mixed-valence V(IV)/V(V) alkoxo-polyoxovanadium cluster series [V6O8(OCH3)11]n+/-: exploring the influence of a mu-oxo ligand in a spin frustrated structure. AB - The synthesis and structural characterization of the neutral mixed-valence methoxo-polyoxovanadium cluster [V(6)O(8)(OCH(3))(11)] (1) and its single oxidation product in the hexachloroantimonate salt [V(6)O(8)(OCH(3))(11)][SbCl(6)] (2) are presented here. The cluster comprises a hexauclear polyoxovanadate core of the Lindqvist structure, of which all but one of the mu-bridging oxo ligands are substituted by methoxo. As revealed by cyclic voltammetry, the cluster is highly redox active, displaying several further thermodynamically stable V(IV)/V(V) mixed-valence redox derivatives. Furthermore, valence sum calculations performed on the X-ray structural data as well as results from IR and UV-vis spectrometry characterize them as class II mixed valence compounds. In the present article, we equally present results from cyclic voltammetry, UV-vis spectrometry, and magnetic measurements obtained for members of the previously reported [V(6)O(7)(OCH(3))(12)] cluster series, which, as opposed to 1 and its derivatives, contain exclusively methoxo ligands as mu bridging moieties. Magnetic measurements performed on the highly reduced cluster species [V(IV)(5)V(V)(1)O(7)(OCH(3))(12)](-) and [V(IV)(6)O(7)(OCH(3))(12)](2-) reveal net antiferromagnetic exchange interactions between the d-electrons, which at lower temperatures are in part suppressed for reasons attributed to geometric spin frustration. Among the present results, the comparison of the cyclic voltammograms of 1 and [V(6)O(7)(OCH(3))(12)] has proven to be of considerable interest, showing an unexpectedly pronounced discrepancy in all but one of their corresponding redox potentials. In particular, a detailed analysis of the electrochemical conversions indicates that the observed shift is almost entirely the result of a different degree of d-electron spin-spin interactions in corresponding mixed valence species of the cluster series. PMID- 19301902 TI - Design of chimeric histone deacetylase- and tyrosine kinase-inhibitors: a series of imatinib hybrides as potent inhibitors of wild-type and mutant BCR-ABL, PDGF Rbeta, and histone deacetylases. AB - Inhibitors of histone deacetylases are a new class of cancer therapeutics with possibly broad applicability. Combinations of HDAC inhibitors with the kinase inhibitor 1 (Imatinib) in recent studies showed additive and synergistic effects. Here we present a new concept by combining inhibition of protein kinases and HDACs, two independent pharmacological activities, in one synthetic small molecule. In general, the HDAC inhibition profile, the potencies, and the probable binding modes to HDAC1 and HDAC6 were similar as for 6 (SAHA). Inhibition of Abl kinase in biochemical assays was maintained for most compounds, but in general the kinase selectivity profile differed from that of 1 with nearly equipotent inhibition of the wild-type and the Imatinib resistant Abl T(315)I mutant. A potent cellular inhibition of PDGFR and cytotoxicity toward EOL-1 cells, a model for idiopathic hypereosinophilic syndrome (HES), are restored or enhanced for selected analogues (12b, 14b, and 18b). Cytotoxicity was evaluated by using a broad panel of tumor cell lines, with selected analogues displaying mean IC(50) values between 3.6 and 7.1 muM. PMID- 19301903 TI - Fishing the target of antitubercular compounds: in silico target deconvolution model development and validation. AB - An in silico target prediction protocol for antitubercular (antiTB) compounds has been proposed in this work. This protocol is the extension of a recently published 'domain fishing model' (DFM), validating its predicted targets on a set of 42 common antitubercular drugs. For the 23 antiTB compounds of the set which are directly linked to targets (see text for definition), the DFM exhibited a very good target prediction accuracy of 95%. For 19 compounds indirectly linked to targets also, a reasonable pathway/embedded pathway prediction accuracy of 84% was achieved. Since mostly eukaryotic ligand binding data was used for the DFM generation, the high target prediction accuracy for prokaryotes (which is an extrapolation from the training data) was unexpected and provides an additional proof of concept of the DFM. To estimate the general applicability of the model, ligand-target coverage analysis was performed. Here, it was found that, although the DFM only modestly covers the entire TB proteome (32% of all proteins), it captures 70% of the proteome subset targeted by 42 common antiTB compounds, which is in agreement with the good predictive ability of the DFM for the targets of the compounds chosen here. In a prospective validation, the model successfully predicted the targets of new antiTB compounds, CBR-2092 and Amiclenomycin. Together, these findings suggest that in silico target prediction tools may be a useful supplement to existing, experimental target deconvolution strategies. PMID- 19301904 TI - Light-induced transfer of molecular chirality in solution: enantiospecific photocyclization of molecularly chiral acrylanilides. AB - Molecularly chiral o-tert-butylacrylanilides undergo enantiospecific 6pi photocyclization to yield 3,4-dihydroquinolin-2-ones with very high enantioselectivity (>90%) in solution. The photocyclization results in the removal of the ortho tert-butyl substituent, presumably via a zwitterionic intermediate. Beta-substitution in the alkene is found to be critical for the transfer of molecular chirality (axial chirality) in the reactant to point chirality in the photoproduct(s). PMID- 19301905 TI - Comparative methods for analysis of protein covalent modification by electrophilic quinoids formed from xenobiotics. AB - Conjugation of biotin and fluorophore tags is useful for assaying covalent protein modification. Oxidative bioactivation of selective estrogen receptor modulators (SERMs) yields reactive quinoid electrophiles that covalently modify proteins, and bioactivation is associated with carcinogenic and chemopreventive effects. Identification of the protein targets of electrophilic metabolites is of general importance for xenobiotics. Four methodologies using SERM derivatized biotin/fluorophore tags were compared for purification and quantification: (1) covert oxidatively activated tags (COATags; SERM conjugated to biotin); (2) dansylTags (SERM conjugated to fluorophore); and azidoTags (SERM azide derivatives) in a two-step conjugation to biotin, using either (3) Staudinger ligation or (4) click chemistry. All synthetic derivatives retained the estrogen receptor ligand characteristics of the parent SERMs. Model proteins with bioactivation by tyrosinase in buffer or cell lysates and liver proteins with in situ bioactivation in rat primary hepatocytes were studied by immunoassay and fluorescence. Comparison showed that the azidoTag/Staudinger method was sensitive but nonspecific, the azidoTag/click methodology had low sensitivity, and the dansylTag methodology failed to detect modified proteins in hepatocytes. The COATag methodology was judged superior, detecting 5 ng of modified protein in vitro and identifying protein targets in hepatocytes. In metabolism studies in rat liver microsomes, the azide group was metabolically labile, which was a contributing factor in not selecting the azidoTag methodology in the oxidative environments required for bioactivation. For study of the protein targets of electrophilic metabolites formed by in situ oxidative bioactivation, the COATag is both sensitive and specific and does not appear to suffer from poor cell permeability. PMID- 19301906 TI - Improvement of chitosan adsorption onto cellulosic fabrics by plasma treatment. AB - Oxygen plasma treatment was applied in order to improve the adsorption of chitosan onto viscose fabric. Modification of the surface and adsorption of chitosan was monitored by determination of XPS spectra, determination of contact angles from rates of water imbibition, and conductometric titration. The plasma treatment resulted in hydrophilization of the surfaces through oxidation. The hydrophilic surfaces were stable for at least 24 h. The treatment also yielded binding sites that resulted in over 20% increase of the amount of chitosan adsorbed over that adsorbed on nontreated fabric. Layers of chitosan adsorbed after plasma treatment were substantially more active as antimicrobial agents than those on nontreated surfaces. PMID- 19301908 TI - MetAlign: interface-driven, versatile metabolomics tool for hyphenated full-scan mass spectrometry data preprocessing. AB - Hyphenated full-scan MS technology creates large amounts of data. A versatile easy to handle automation tool aiding in the data analysis is very important in handling such a data stream. MetAlign softwareas described in this manuscripthandles a broad range of accurate mass and nominal mass GC/MS and LC/MS data. It is capable of automatic format conversions, accurate mass calculations, baseline corrections, peak-picking, saturation and mass-peak artifact filtering, as well as alignment of up to 1000 data sets. A 100 to 1000-fold data reduction is achieved. MetAlign software output is compatible with most multivariate statistics programs. PMID- 19301907 TI - Personalized metabolic assessment of erythrocytes using microfluidic delivery to an array of luminescent wells. AB - The metabolic syndrome is often described as a group of risk factors associated with diabetes. These risk factors include, but are not limited to, such conditions as insulin resistance, obesity, high blood pressure, and oxidant stress. Here, we report on a tool that may provide some clarity on the relationship between some of these associated risk factors, especially oxidant stress and hypertension. Specifically, we describe the ability to simultaneously monitor nicotinamide dinucleotide phosphate (NADPH), reduced glutathione (GSH), and shear-induced adenosine triphosphate (ATP) release from erythrocytes using luminescence detection on a microfabricated device. The measurements are performed by delivering erythrocyte lysate (for the NADPH and GSH measurements, two analytes indicative of oxidative stress) or whole red blood cells (RBCs) (for the determination of ATP release from the cells) to an array of wells that contain the necessary reagents to generate a luminescence emission that is proportional to analyte concentration. A fluorescence macrostereomicroscope enables whole-chip imaging of the resultant emission. The concentrations of each NADPH and GSH contained within a 0.7% erythrocyte solution were determined to be 31.06 +/- 4.12 and 22.55 +/- 2.47 microM, respectively, and the average ATP released from a nonlysed 7% erythrocyte solution was determined to be 0.54 +/- 0.04 microM. Collectively, the device represents a precursor to subsequent merging of microfluidics and microtiter-plate technology for high-throughput assessment of metabolite profiles in the diabetic erythrocyte. PMID- 19301909 TI - Modulation of protein-surface interactions on nanopatterned polymer films. AB - The introduction of nanoscale features brings with it a high density of surface interface boundaries and effectively introduces an additional boundary material that exhibits properties different from the surrounding surfaces. We systematically varied the feature size of self-assembled polystyrene-block poly(methyl methacrylate) copolymer nanopatterns from 13 to 200 nm and demonstrated that the basic property of protein adsorption on a nanopatterned surface can be modulated by the length density of surface interfaces present. Protein adsorption on the nanopatterns could be described by a modified adsorption affinity along the surface interface with an effective width on the length-scale of individual proteins. Due to the intrinsic high density of surface interfaces in many polymeric thin film nanopatterns and structures, the interaction of proteins with such interfaces may be of particular relevance to cell-surface studies and to biomaterial and biosensor applications involving nanoscale features. PMID- 19301910 TI - Quantification of antioxidant capacity in a microemulsion system: synergistic effects of chlorogenic acid with alpha-tocopherol. AB - We report herein a characterization of an oil-in-water (o/w) microemulsion consisting of 12% methyl linoleate (or styrene in weight %), 29% surfactant Tween 20, 15% n-butanol, and 44% 75 mM potassium phosphate buffer (pH 7.04). The oil phase droplet size, determined by dynamic light scattering, is 19.3 nm with polydispersity at 0.103. When methyl linoleate is replaced with styrene, the droplet size increases to 63 nm but with much narrower polydispersity at 0.047. The droplet size of styrene is confirmed by polymerization of the styrene in the microemulsion. The polystyrene particles isolated have diameters of approximately 70 nm determined by a scanning electronic microscope. Both microemulsions remain stable for two months as expected for a thermodynamically stable system. The methyl linoleate oxidation is induced by AAPH, and the effects of radical scavengers are evaluated in a high throughput fashion using an oxygen sensor coated 96-well microplate Oxygen Biosensor Systems. From the oxygen consumption kinetic curves, the antioxidant capacity can be calculated using Trolox as the standard. The synergistic effect of hydrophilic antioxidants and alpha-tocopherol was measured for representative flavonoids. Chlorogenic acid has the best synergistic effect of 44.8%. The assay, coined as ORAC(E) standing for oxygen radical absorbance capacity in microemulsion, provides a model system in evaluating antioxidant capacity of phenolic compounds in a heterogeneous system relevant to food and cosmetic applications. PMID- 19301911 TI - Substrate induced population shifts and stochastic gating in the PBCV-1 mRNA capping enzyme. AB - The 317 residue PBCV-1 mRNA capping enzyme catalyzes the second enzymatic reaction in the formation of an N-7-methyl-GMP cap on the 5'-end of the nascent mRNA. It is composed of two globular domains bound by a short flexible peptide linker, which have been shown to undergo opening and closing events. The small size and experimentally demonstrated domain mobility make the PBCV-1 capping enzyme an ideally suited model system to explore domain mobility in context of substrate binding. Here, we specifically address the following four questions: (1) How does substrate binding affect relative domain mobility: is the system better described by an induced fit or population shift mechanism? (2) What are the gross characteristics of a conformation capable of binding substrate? (3) Does "domain gating" of the active site affect the rate of substrate binding? (4) Does the magnitude of receptor conformational fluctuations confer substrate specificity by sterically occluding molecules of a particular size or geometry? We answer these questions using a combination of theory, Brownian dynamics, and molecular dynamics. Our results show that binding efficiency is a function of conformation but that isomerization between efficient and inefficient binding conformations does not impact the substrate association rate. Additionally, we show that conformational flexibility alone is insufficient to explain single stranded mRNA specificity. While our results are specific to the PBCV-1 mRNA capping enzyme, they provide a useful context within which the substrate binding behavior of similarly structured enzymes or proteins may be considered. PMID- 19301912 TI - A mechanistic study of S(N)2 reaction in a diol solvent. AB - We present calculations for the mechanism of S(N)2 reactions in ethylene glycol, focusing on the role of two protic functional groups (-OH) in the solvent molecule. We find that some hydroxyl groups act as Lewis base to interact with the cation, whereas others affect the reaction as Lewis acid to the nucleophile. We predict that the nucleophile (F-) reacts as an ion pair rather than as a solvent-separated ion when metal cation (Cs+) is used as a counterion. The overall influence of ethylene glycol manifests itself as the reaction barrier (E(double dagger) = 20.0, G(353K)(double dagger) = 21.5 kcal/mol) that is a bit smaller than that in tert-butyl alcohol, which proved to be a very efficient solvent for S(N)2 reactions [Kim, D. W., et al. J. Am. Chem. Soc. 2006, 128, 16394]. We therefore show that a small protic solvent such as ethylene glycol may be as efficient as a bulky alcohol for S(N)2 reactions. PMID- 19301913 TI - The unfolded state of the C-terminal domain of the ribosomal protein L9 contains both native and non-native structure. AB - Interest in the structural and dynamic properties of unfolded proteins has increased in recent years owing to continued interest in protein folding and misfolding. Knowledge of the unfolded state under native conditions is particularly important for obtaining a complete picture of the protein folding process. The C-terminal domain of protein L9 is a globular alpha, beta protein with an unusual mixed parallel and antiparallel beta-strand structure. The folding kinetics and equilibrium unfolding of CTL9 strongly depend on pH, and follow a simple two state model. Both the native and the unfolded state can be significantly populated at pH 3.8 in the absence of denaturant, allowing the native state and the unfolded state to be characterized under identical conditions. Backbone (15)N, (13)C, (1)H and side-chain (13)C(beta), (1)H(beta) chemical shifts, amide proton NOEs, and (15)N R(2) relaxation rates were obtained for the two conformational states at pH 3.8. All the data indicate that the pH 3.8 native state is well folded and is similar to the native state at neutral pH. There is significant residual structure in the pH 3.8 unfolded state. The regions corresponding to the two native state alpha-helices show strong preference to populate helical phi and psi angles. The segment that connects alpha-helix 2 and beta-strand 2 has a significant tendency to form non-native alpha-helical structure. Comparison with the pH 2.0 unfolded state and the urea unfolded state indicates that the tendency to adopt both native and non-native helical structure is stronger at pH 3.8, demonstrating that the unfolded state of CTL9 under native like conditions is more structured. The implications for the folding of CTL9 are discussed. PMID- 19301914 TI - Effect of gas pressure and gas type on the fragmentation of peptide and oligosaccharide ions generated in an elevated pressure UV/IR-MALDI ion source coupled to an orthogonal time-of-flight mass spectrometer. AB - Matrix-assisted laser desorption ionization (MALDI) allows for the mass spectrometric (MS) analysis of thermally labile, non-volatile biomolecules. However, some residual analyte fragmentation typically accompanies the phase transition from the condensed to the gas phase and following plume expansion, even under optimized conditions. In-source decay (ISD) and post-source decay (PSD) MALDI MS are two techniques that make use of these phenomena and that can provide useful structural information by producing characteristic fragment ions of the analyte compounds. In orthogonal extracting time-of-flight mass spectrometry (o-TOF-MS), the pressure of the cooling gas in the ion source has a strong influence on the extent of analyte ion fragmentation. We investigated the effect of this parameter on peptide and oligosaccharide fragmentation by examining a range of pressures (from 0.05-1.8 mbar) in combination with seven different buffer gases (He, Ne, Ar, N(2), CO(2), CH(3), isobutane). Ions were generated by ultraviolet (UV) and/or by infrared (IR) MALDI. The influence of the ion extraction voltage on the analyte fragmentation also was investigated for a selected set of gas parameters. We observed that individual fragment ions exhibit characteristic fragment yield-pressure dependencies that can be classified into three groups. Type I ions resemble species that are also found in MALDI PSD MS analysis, while type II ions resemble typical ISD fragments. The yield-pressure relationship of type III ions suggests that these are the result of a combination of both processes. Comparing the yields of fragmentation for the different buffer gases reveals a correlation between their internal degrees of freedom and their collisional cooling efficiency. Changing the buffer gas pressure and/or extraction field provides an easy means to influence analyte ion fragmentation and to switch from the primary production of one type of fragment species to another. The method can therefore facilitate the structural characterization of MALDI-generated ions. PMID- 19301915 TI - Elucidation of functions of human cytochrome P450 enzymes: identification of endogenous substrates in tissue extracts using metabolomic and isotopic labeling approaches. AB - One of the central problems in biochemistry in the postgenomic era is the elucidation of functions of proteins, including "orphan" human cytochromes P450 (P450s), when the substrates are unknown. A general strategy for identification of endogenous substrates of P450s in tissue extracts using metabolomic and isotopic labeling approaches is described, involving four main steps: (1) In vitro incubation of a P450 enzyme system with cofactor and tissue extract is done under a mixture of (18)O(2)/(16)O(2) (1:1). (2) Liquid chromatography/mass spectrometry (LC/MS) assay of an organic extract of the reaction mixture is performed. (3) The isotopic labeling products appearing as M/M + 2 doublets can be directly identified using the program DoGEX (Sanchez-Ponce, R. and Guengerich, F. P. Anal. Chem. 2007, 79, 3355-3362). (4) Characterization of potential candidates is done. Validation of the strategy was established using human P450 7A1 as an initial model to identify its known product, 7alpha-hydroxycholesterol, in liver extracts. The strategy was then applied to human P450s 1A2, 2C8, and 2C9 in untargeted substrate searches with human liver extracts. A total of seven fatty acids were identified and verified as substrates of these three hepatic P450s. The products were subsequently characterized as hydroxylation and epoxidation derivatives of fatty acids, using gas chromatography/mass spectrometry (GC/MS) analysis. Finally, kinetic studies were performed to confirm that the fatty acids are oxidized by P450s 1A2, 2C8, and 2C9. Thus, this strategy has been demonstrated to be useful in identifying reactions in tissue extracts with orphan human P450s. PMID- 19301916 TI - Synthesis and characterization of iron derivatives having a pyridine-linked bis(anilide) pincer ligand. AB - A pyridine-linked bis(aniline) pincer ligand, [2]H(2) ([2]H(2) = (2,6-NC(5)H(3)(2 (2,4,6-Me(3)C(6)H(2))-NHC(6)H(4))(2)), has been synthesized in two steps. Deprotonation with Me(3)SiCH(2)Li followed by metalation with FeCl(2) yielded a LiCl adduct of [2]Fe. The complex is freed of LiCl with excess TlPF(6) or by crystallization from toluene/petroleum ether, giving [2]Fe(THF). [2]Fe(THF) reacts with I(2) and O(2) to generate [2]FeI and ([2]Fe)(2)O, respectively. The complexes have been characterized by (1)H NMR spectroscopy, elemental analysis, X ray crystallography, and UV-vis spectroscopy. [2]Fe(THF) has been examined using cyclic voltammetry. PMID- 19301917 TI - DFT study on N2 activation by a hydride-bridged diniobium complex. N[triple bond]N bond cleavage accompanied by H2 evolution. AB - Density functional theory (DFT) calculations have been performed for the investigation of a plausible mechanism of the triple bond cleavage of N(2) in a diniobium complex supported by tridentate aryloxide ligands, {Nb(V)(mu N)(2)Nb(V)}(2-). (22) With the assumption of a tetrakis(mu-hydrido)diniobium complex {Nb(IV)(mu-H)(4)Nb(IV)}(2-) as an initial complex, the NN cleavage on the Nb(2) core proceeds in four steps. Dinitrogen is coordinated to the {Nb(III)(mu H)(2)Nb(III)} core in a side-on/end-on manner, accompanied by the reductive elimination of H(2). The N[triple bond]N bond of dinitrogen is activated up to a single bond (formally N(2)(4-)) by the two Nb(III) atoms, once it is bound to the Nb(2) core. Two electrons are prepared for the cleavage of the N-N single bond through the mu-H migration to an N atom, leading to the formation of an Nb-Nb bond. The N-N bond is then dissociated by the two electrons that are shared between the two Nb atoms. Finally, {Nb(mu-N)(2)Nb}(2-) is generated after H(2) elimination in which the N-bonded H atom is coupled with the remaining mu-H atom. The final H(2) elimination is calculated to be the rate-determining step. PMID- 19301918 TI - The toxicity of prion protein fragment PrP(106-126) is not mediated by membrane permeabilization as shown by a M112W substitution. AB - Prion diseases result from a post-translational modification of the physiological prion protein (PrP(C)) into a scrapie isoform (PrP(Sc)). The PrP(106-126) fragment is conserved among various abnormal variants and shows PrP(Sc) pathogenic properties. It has been proposed that the PrP(106-126) fragment may exhibit its toxic effects through membrane pore formation. Our previous studies showed that PrP(106-126) does not interact with membranes under physiological conditions. In the present study, PrP(106-126) affinity for membranes was increased by modifying PrP(106-126) with a M112W substitution, and pore formation was further evaluated. However, while the peptide exhibited an increased local concentration in the membrane, this did not lead to the induction of membrane permeabilization, as verified by fluorescence methodologies and surface plasmon resonance. These results further support the idea that PrP(106-126) toxicity is not a consequence of peptide-membrane interaction and pore formation. PMID- 19301919 TI - Tuning the framework topologies of Co(II)-doped Zn(II)-tetrazole-benzoate coordination polymers by ligand modifications: structures and spectral studies. AB - In our continuing efforts to explore the effects of ligand modifications on the structures and properties of their metal complexes, we studied the in situ [2 + 3] cycloaddition reactions of benzonitrile, o-phthalodinitrile, 3-cyanobenzoic acid, 4-cyanobenzoic acid with NaN(3) in the presence of Zn(II) and/or Co(II) salts under hydrothermal conditions, and obtained four new Co(II)-doped Zn(II) tetrazole-benzoate coordination polymers with the formula of [Co(x)Zn(1 x)(L(m))(y)](n) [5-phenyl-1H-tetrazole (HL(1)) for 1, 2-(1H-tetrazol-5-yl)benzoic acid (H(2)L(2)) for 2, 3-(1H-tetrazol-5-yl)benzoic acid (H(2)L(3)) for 3, and 4 (1H-tetrazol-5-yl)benzoic acid (H(2)L(4)) for 4]. The structure of 1 shows a classical diamondoid net, while 2 and 3, take 2D layer structure with (4.6(2))(4.6.4.6) topology and 3D SrAl(2) topology, respectively. The structure of 4 exhibits a four-connected 3D network with rare non-diamondoid 6(6) topology. The coordination modes of the center metal and the ligands in the four complexes are almost the same, being tetrahedral or four-connected, but their topologies are quite different. Thus, the four structurally related ligands allow analysis of the effects of the disposition of a second functional carboxylate group on an aromatic ring and the twist angles of the carboxylate and tetrazoyl out of the plane of the aromatic ring on the overall structural topology of their complexes. Interestingly, the Co(II) ions were doped into the Zn(II) complexes, as confirmed by their macroscopical colors, inductively coupled plasma (ICP) analysis and UV visible spectra. In addition, the photoluminescence of the four complexes in the solid state at room temperature was briefly studied. PMID- 19301920 TI - Nanostructure initiator mass spectrometry: tissue imaging and direct biofluid analysis. AB - Nanostructure initiator mass spectrometry (NIMS) is a recently introduced matrix free desorption/ionization platform that requires minimal sample preparation. Its application to xenobiotics and endogenous metabolites in tissues is demonstrated, where clozapine and N-desmethylclozapine were observed from mouse and rat brain sections. It has also been applied to direct biofluid analysis where ketamine and norketamine were observed from plasma and urine. Detection of xenobiotics from biofluids was made even more effective using a novel NIMS on-surface extraction method taking advantage of the hydrophobic nature of the initiator. Linear response and limit of detection were also evaluated for xenobiotics such as methamphetamine, codeine, alprazolam, and morphine, revealing that NIMS can be used for quantitative analysis. Overall, our results demonstrate the capacity of NIMS to perform sensitive, simple, and rapid analyses from highly complex biological tissues and fluids. PMID- 19301921 TI - Structure revision of the lantibiotic 97518. AB - The lantibiotic 97518, produced by a Planomonospora sp., was reported as a 2194 Da polypeptide comprising 24 amino acid residues with five thioether bridges. It was assigned to the mersacidin subgroup of type B lantibiotics by Castiglione et al. (Biochemistry 2007, 46, 5884-5897) and named planosporicin. New analytical, chemical, and genetic data and reinterpretation of the published NMR chemical shifts enable structure revision of 97518. The resulting revision of the 97518 structure involves both a shift of two amino acids and a reorganization of two thioether bridges. With this revision, the lantibiotic 97518 becomes a clear member of the nisin subgroup of compounds. PMID- 19301922 TI - An unusual pathway to cyclobutane formation via desulfurative intramolecular photocycloaddition of an enone benzothiazoline pair. AB - Irradiation of the enone benzothiazoline 3 leads to the formation of cyclobutane 5. Preliminary mechanistic studies establish the intermediacy of an enecarbamate 14 in this photochemical transformation, which could be the result of sulfur extrusion from an episulfide intermediate. Photocycloaddition of the enecarbamate intermediate 14 leads to the formation of "crossed" photoadducts, i.e., 5, in excellent yield, with high levels of regio- and stereochemical control. PMID- 19301923 TI - Nano indium oxide as a recyclable catalyst for C-S cross-coupling of thiols with aryl halides under ligand free conditions. AB - An efficient ligand-free C-S cross-coupling of aryl halides with aromatic/alkyl thiols has been developed using a catalytic amount of nanocrystalline indium oxide as a recyclable catalyst with KOH as the base in DMSO at 135 degrees C. A variety of aryl sulfides can be synthesized in excellent yields utilizing this protocol. PMID- 19301924 TI - Multicomponent type II anion relay chemistry (ARC): one-pot syntheses of 2,3 disubstituted furans and thiophenes. AB - Effective, one-pot syntheses of 2,3-disubstituted furans and thiophenes, exploiting 2-tert-butyldimethylsilyl-3-formylfuran and -thiophene as the respective bifunctional linchpins, have been developed. The synthetic protocol involves multicomponent type II Anion Relay Chemistry (ARC) mediated by a solvent controlled C(sp(2))-->O 1,4-Brook rearrangement. Simple organolithiums and alpha disubstituted ester enolates prove effective as the initiating nucleophiles. PMID- 19301925 TI - Nickel-catalyzed direct arylation of azoles with aryl bromides. AB - Nickel catalyst systems for the direct C2 arylation of oxazoles and thiazoles have been developed. The catalyst systems are cost-efficient and allow the use of various aryl bromides in the C-H arylation of azoles. PMID- 19301926 TI - Exposure of epitaxial graphene on SiC(0001) to atomic hydrogen. AB - Graphene films on SiC exhibit coherent transport properties that suggest the potential for novel carbon-based nanoelectronics applications. Recent studies suggest that the role of the interface between single layer graphene and silicon terminated SiC can strongly influence the electronic properties of the graphene overlayer. In this study, we have exposed the graphitized SiC to atomic hydrogen in an effort to passivate dangling bonds at the interface, while investigating the results utilizing room temperature scanning tunneling microscopy. PMID- 19301927 TI - Building molecular complexity via tandem Ru-catalyzed isomerization/C-H activation. AB - A tandem isomerization/C-H activation of allylic alcohols was performed using a catalytic amount of RuCl(2)(PPh(3))(3). A variety of ortho alkylated ketones have been obtained in excellent yields. This tandem process relies on an in situ generation of a carbonyl functional group that directs the ortho C-H bond activation. PMID- 19301928 TI - Kinetic resolution of indolines by pd-catalyzed asymmetric allylic amination. AB - The kinetic resolution of indolines was realized via a Pd-catalyzed allylic substitution reaction by using Trost's chiral ligand L10, affording optically active indolines and N-allylated indolines in high yields and high enantioselectivities with an S factor up to 59, which provided the first example for the kinetic resolution of nucleophiles via a transition-metal-catalyzed allylic substitution reaction. PMID- 19301929 TI - Conformational distribution and ultrafast base dynamics of leadzyme. AB - The dynamic nature of ribozymes represents a significant challenge in elucidating their structure-dynamics-function relationship. Here, using femtosecond time resolved spectroscopy and other biophysical tools, we demonstrate that the active site of leadzyme does not have a unique structure, but rather samples an ensemble of conformations that undergo picosecond structural changes. Various base modifications have a profound context-dependent impact on the catalysis. PMID- 19301930 TI - Interplay between covalent and physical interactions within environment sensitive hydrogels. AB - A systematic study is carried out to understand how physical and covalent crosslinks affect the mechanical properties of an eight-arm poly(ethyleneglycol) based hydrogel. Heparin and heparin-binding peptide are used as a physical crosslinker, and an enzymatically cleavable peptide with a cysteine on each end serves as a covalent crosslinker. While physical crosslinks alone do not induce gelation due to the low binding affinity between heparin and heparin-binding peptide, the addition of covalent crosslinks leads to gel formation. Strikingly, the addition of the covalent crosslinks not only leads to gel formation, but also enhances the contribution from the physical crosslinks to the overall shear moduli, which are negligible in the absence of covalent crosslinks. The gels, which contain both covalent and physical crosslinks, are able to reversibly respond to external stimuli such as temperature and oscillatory shear unlike the purely covalent gel in which the moduli remain largely insensitive to such stimuli. Two explanations are provided for this striking phenomenon. First, the addition of covalent crosslinks increased the stress relaxation time of the gel enabling the physical interactions to contribute to the moduli. This is contrasted to the case of physically crosslinked material, which relaxes the stress too quickly, preventing the physical interactions from contributing to the low frequency moduli. Second, it is believed that the physical interactions within the covalent network were further enhanced by "macromolecular confinement", which favors the formation of compact conformational structures in the confined space. Quartz crystal microbalance (QCM) was used to measure the dissociation constant (K(d)) within the hydrogel and to demonstrate that the binding between heparin and heparin-binding peptide is stronger within the gel compared to that within the solution phase. Because extracellular matrix (ECM) contains both covalent and physical interactions between its constituents, and the mechanical properties of the ECM are important factors to control cell functions, the findings of this research may have important implications in various fields of tissue engineering and cell biology. PMID- 19301931 TI - Anti-diarrheal effects of diosmectite in the treatment of acute diarrhea in children: a review. AB - Acute diarrhea caused by an infectious organism is highly prevalent, particularly in developing countries, and is usually self-limiting. The condition is a major contributor to the global disease burden and is associated with a significant economic cost. The disease is common in children aged <5 years. It occurs as the result of exposure to a diarrheogenic agent that alters intestinal absorption and/or secretion, resulting in an increase in the volume of water that enters the colon beyond that which can be absorbed. Diarrhea almost always occurs by one or more of four mechanisms: disruption of osmotic forces in the intestine; disruption of normal secretory processes; disruption of epithelial cells or the epithelial tight junctions; or motility disorders. Oral rehydration therapy (ORT) is central to the management of acute diarrhea, and is sufficient to prevent complications due to dehydration in most patients while the disease runs its course. However, ORT has no effect on the duration of the disease or frequency of bowel motions, and any agent that could meet these needs would therefore be a useful addition to ORT. Diosmectite is a natural aluminomagnesium silicate clay with a lamellar, non-fibrous crystalline structure that gives it strong adsorbent properties. Its mechanisms of action are not yet fully understood, but are probably multiple. Diosmectite reduces inflammation, modifies mucus rheologic properties, inhibits mucolysis, and adsorbs bacteria, bacterial enterotoxins, viruses and other potentially diarrheogenic substances. A number of studies have shown that diosmectite reduces the duration of diarrhea and decreases the frequency of bowel motions after 2 days of treatment in children with mild-to moderate acute diarrhea. Two recent trials have indicated a reduction in stool output with diosmectite. Because of its very favorable safety profile, with no serious adverse effects, diosmectite is frequently used, mainly in European countries but also in Asia and Africa. Diosmectite should be used as an adjunct, not an alternative, to ORT and, when needed, to antibacterial therapy. PMID- 19301932 TI - The role of inhaled corticosteroids in the management of cystic fibrosis. AB - The lung disease of cystic fibrosis (CF) is characterized by a vicious cycle of airway obstruction, chronic bacterial infection, and vigorous inflammation, which ultimately results in bronchiectasis. Recognition that excessive and persistent inflammation is a key factor in lung destruction has prompted investigation into anti-inflammatory therapies. Although effective, the use of systemic corticosteroids has been limited by the unacceptable adverse effect profile. Inhaled corticosteroids (ICS) are a widely prescribed anti-inflammatory agent in CF, likely as a result of clinicians' familiarity with these agents and their excellent safety profile at low doses in asthmatic patients. However, while multiple studies are limited by small sample size and short duration, they consistently failed to demonstrate statistically or clinically significant benefits of ICS use in CF. This review provides an overview of the inflammatory response in CF, the mechanisms of action of corticosteroids, the safety of ICS, and the literature relevant to the use of ICS in CF. PMID- 19301933 TI - Etiology, diagnosis, and pharmacologic treatment of pediatric pulmonary hypertension. AB - Major advances have been made in the understanding and treatment of pulmonary hypertension in the last few years. Without treatment (medication) for idiopathic pulmonary arterial hypertension, which is a rare and potentially fatal condition, the survival time is only about 3 years after diagnosis. However, if pulmonary hypertension is secondary to other causes such as congenital heart disease, it is possible to survive for 30 years or more without treatment. The condition can affect children at any age, from fetal life to adulthood. Patients with pulmonary hypertension can present to the respiratory pediatrician with unresponsive asthma, to the neurologist with faints, or to the general pediatrician with failure to thrive. Over the last few years there have been significant developments in the available therapy for managing this complicated disease. There is now a generally recognized ladder of long-term therapy for chronic pulmonary hypertension. Treatment can start with oxygen at home at night or even during the day. Next is the use of oral phosphodiesterase inhibitors, mostly type V, such as sildenafil, which enhance endogenous nitric oxide. More potent are the endothelin receptor antagonists and the most potent are the prostanoids, especially epoprostenol, which is given by constant intravenous infusion. In addition to interventional catheterization with atrial septostomy, these agents have improved the prognostic outlook. This article reviews the current knowledge about the etiology, investigation, and treatment of children with pulmonary hypertension in the clinical setting. PMID- 19301935 TI - Incidence, preventability, and impact of Adverse Drug Events (ADEs) and potential ADEs in hospitalized children in New Zealand: a prospective observational cohort study. AB - Adverse drug events (ADEs) are an important problem in all hospitalized patients as these events represent medication-related patient harm. Few epidemiologic data exist regarding ADEs in the pediatric inpatient setting and, in particular, the economic impact of such ADEs upon the healthcare sector. To evaluate the incidence, preventability, and seriousness of ADEs and potential ADEs occurring in hospitalized children and to examine the cost implications of these ADEs. This was a prospective observational cohort study conducted in the pediatric, neonatal intensive care unit (NICU), and postnatal wards of a university-affiliated urban general hospital in Dunedin, New Zealand (NZ). The study population was all patients admitted to these wards for >24 hours over a 12-week period from 18 March 2002 to 9 June 2002. Medication-related events were identified by chart review, attendance at multidisciplinary clinical meetings, parent/carer/child interviews, and voluntary and verbally solicited reports from staff. All suspected medication-related events were reviewed by a panel of three health professionals who independently categorized the events and rated them for seriousness, preventability, and causality, using a standardized reviewer form. Costs attributable to ADEs were calculated using both the average cost of a bed day, and specific costs for diagnostic groupings. The main outcome measures of the study were ADEs and potential ADEs. There were 495 eligible study patients, who had a total of 520 admissions and 3037 patient-days of admission, during which 3160 prescription episodes were written. There were 67 ADEs, of which 38 (56.7%) were classified as preventable, and 77 potential ADEs. ADEs occurred at a rate of 2.1 per 100 prescription episodes, 12.9 per 100 admissions, and 22.1 per 1000 patient-days. Potential ADEs occurred at a rate of 2.4 per 100 prescription episodes, 14.6 per 100 admissions, and 25 per 1000 patient-days. Although the greatest number (and rate per 100 admissions) of ADEs occurred in NICU patients, surgical pediatric ward patients had the greatest rate of ADEs per 1000 patient days. Few events occurred in postnatal patients. Forty-six percent of ADEs were classified as being serious; 15% were deemed to result in persistent disability or were classified as life threatening. Potential ADEs were deemed more likely to be serious with 82% classified as potentially serious events; 33% were deemed as having the potential to result in persistent disability, or the potential to cause a life-threatening event. Fifteen ADEs were judged to have caused the hospital admission or to have prolonged hospital stay. The total number of days attributed to ADEs was 92 (range 1-26 days); of these, 58 were deemed preventable days and 34 non-preventable days. This extrapolates to a total annual cost of $NZ235 214 (2002 values) to the pediatric service, subdivided into $NZ148 287 for preventable ADEs and $NZ86 927 for non-preventable ADEs. ADEs and potential ADEs represent a considerable hazard for the pediatric inpatient population and ADEs represent a large cost imposition upon the healthcare sector. Over half of the ADEs were deemed preventable. This highlights the importance of developing strategies to prevent and ameliorate ADEs both to improve the quality of patient care and to reduce healthcare costs. PMID- 19301934 TI - Pediatric palliative care: use of opioids for the management of pain. AB - Pediatric palliative care (PPC) is provided to children experiencing life limiting diseases (LLD) or life-threatening diseases (LTD). Sixty to 90% of children with LLD/LTD undergoing PPC receive opioids at the end of life. Analgesia is often insufficient. Reasons include a lack of knowledge concerning opioid prescribing and adjustment of opioid dose to changing requirements. The choice of first-line opioid is based on scientific evidence, pain pathophysiology, and available administration modes. Doses are calculated on a bodyweight basis up to a maximum absolute starting dose. Morphine remains the gold standard starting opioid in PPC. Long-term opioid choice and dose administration is determined by the pathology, analgesic effectiveness, and adverse effect profile. Slow-release oral morphine remains the dominant formulation for long-term use in PPC with hydromorphone slow-release preparations being the first rotation opioid when morphine shows severe adverse effects. The recently introduced fentanyl transdermal therapeutic system with a drug-release rate of 12.5 microg/hour matches the lower dose requirements of pediatric cancer pain control. Its use may be associated with less constipation compared with morphine use. Though oral transmucosal fentanyl citrate has reduced bioavailability (25%), it inherits potential for breakthrough pain management. However, the gold standard breakthrough opioid remains immediate-release morphine. Buprenorphine is of special clinical interest as a result of its different administration routes, long duration of action, and metabolism largely independent of renal function. Antihyperalgesic effects, induced through antagonism at the kappa-receptor, may contribute to its effectiveness in neuropathic pain. Methadone also has a long elimination half-life (19 [SD 14] hours) and NMDA receptor activity although dose administration is complicated by highly variable morphine equianalgesic equivalence (1 : 2.5-20). Opioid rotation to methadone requires special protocols that take this into account. Strategies to minimize adverse effects of long-term opioid treatment include dose reduction, symptomatic therapy, opioid rotation, and administration route change. Patient- or nurse-controlled analgesia devices are useful when pain is rapidly changing, or in terminal care where analgesic requirements may escalate. In this article, we present detailed pediatric pharmacokinetic and pharmacodynamic data for opioids, their indications and contraindications, as well as dose-administration regimens that include practical strategies for opioid switching and dose reduction. Additionally, we discuss the problem of hyperalgesia and the use of adjuvant drugs to support opioid therapy. PMID- 19301936 TI - Flushing ASsessment Tool (FAST): psychometric properties of a new measure assessing flushing symptoms and clinical impact of niacin therapy. AB - BACKGROUND AND OBJECTIVE: A common adverse effect of niacin therapy is flushing, manifested by cutaneous warmth, redness, itching and/or tingling. The Flushing ASsessment Tool (FAST) was developed to assess flushing symptoms and their impact on patients receiving niacin therapy. This study evaluated the reliability, validity and responsiveness of the FAST. The minimal important difference (MID) of the FAST was also examined. METHODS: This was a prospective, randomized, double-blind, placebo-controlled, parallel-group 8-week study conducted to evaluate the psychometric characteristics of the FAST. The instrument is administered daily using an electronic patient diary. The study was conducted at 41 clinical sites in the US. 276 patients with dyslipidaemia were randomized to treatment and were at least 18 years of age, with fasting laboratory values of low-density lipoprotein cholesterol (LDL-C) <250 mg/dL and one of the following: high-density lipoprotein cholesterol (HDL-C) <40 mg/dL for males or <50 mg/dL for females; or triglycerides (TG) > or = 150 and < or = 400 mg/dL; or LDL-C > or = 70 mg/dL for patients with a history of coronary heart disease (CHD) or CHD risk equivalents, or > or = 100 mg/dL for subjects with two risk factors, or > or = 160 mg/dL for subjects with 0-1 risk factors. Patients were randomized (1 : 1 : 1) to receive niacin extended-release (NER) 500 mg/day in week 1, 1000 mg/day in week 2 and 2000 mg/day in weeks 3-6/aspirin (acetylsalicylic acid [ASA]), NER/ASA placebo, or NER placebo/ASA placebo. RESULTS: FAST test-retest reliability in stable patients during the first 2 weeks was demonstrated for overall flushing severity using patient and physician overall treatment effect (OTE) ratings (intraclass correlation coefficients of >0.7 for mean overall and individual flushing severity scores). Over the 6-week treatment period, FAST scores demonstrated significant correlations with individual symptoms, impact on daily activities and sleep, and dissatisfaction related to flushing (p < 0.01). Changes in FAST scores were associated with treatment satisfaction (p < 0.01) and patient and physician-rated OTE (p < 0.01). Using patient-rated OTE, the mean maximum flushing severity scores improved 1.85 points in responders and only 0.18 points in non-responders (p < 0.001); responders were defined by improved patient- or physician-rated OTE. Among patients with flushing, mean maximum overall flushing scores differed between patients who subsequently discontinued due to flushing (7.9 points) and those who did not discontinue (4.7 points; p < 0.001). The probable range in this study for a detectable change in flushing symptoms (MID) was 0.29-0.38 points for mean flushing severity and 0.66-0.86 points for maximum flushing severity. CONCLUSION: The FAST exhibited test-retest reliability, good evidence of construct validity, and, overall, flushing severity was responsive to change over time. The FAST is a reliable and valid instrument for assessing the impact of niacin-induced flushing in patients with dyslipidaemia. PMID- 19301937 TI - Efficacy and safety of 5% lidocaine (lignocaine) medicated plaster in comparison with pregabalin in patients with postherpetic neuralgia and diabetic polyneuropathy: interim analysis from an open-label, two-stage adaptive, randomized, controlled trial. AB - BACKGROUND AND OBJECTIVE: Postherpetic neuralgia (PHN) and diabetic polyneuropathy (DPN) are two common causes of peripheral neuropathic pain. Typical localized symptoms can include burning sensations or intermittent shooting or stabbing pains with or without allodynia. Evidence-based treatment guidelines recommend the 5% lidocaine (lignocaine) medicated plaster or pregabalin as first-line therapy for relief of peripheral neuropathic pain. This study aimed to compare 5% lidocaine medicated plaster treatment with pregabalin in patients with PHN and patients with DPN. METHODS: The study was a two-stage, adaptive, randomized, controlled, open-label, multicentre trial that incorporated a drug wash-out phase of up to 2 weeks prior to the start of the comparative phase. At the end of the enrollment phase, patients who fulfilled the eligibility criteria were randomized to either 5% lidocaine medicated plaster or pregabalin treatment and entered the 4-week comparative phase. The interim analysis represents the first stage of the two-stage adaptive trial design and was planned to include data from the comparative phase for the first 150 randomized patients of the 300 total planned for the trial. Patients aged > or = 18 years with PHN or DPN were recruited from 53 investigational centres in 14 European countries. For this interim analysis, 55 patients with PHN and 91 with DPN (full-analysis set [FAS]), randomly assigned to the treatment groups, were available for analysis. Topical 5% lidocaine medicated plaster treatment was administered by patients to the area of most painful skin. A maximum of three or four plasters were applied for up to 12 hours within each 24-hour period in patients with PHN or DPN, respectively. Pregabalin capsules were administered orally, twice daily. The dose was titrated to effect: all patients received 150 mg/day in the first week and 300 mg/day in the second week of treatment. After 1 week at 300 mg/day, the dose of pregabalin was further increased to 600 mg/day in patients with high pain intensity scores. The pre-planned primary study endpoint was the rate of treatment responders, defined as completing patients experiencing a reduction from baseline of > or = 2 points or an absolute value of < or = 4 points on the 11-item numerical rating scale of recalled average pain intensity over the last 3 days (NRS-3), after 4 weeks of treatment. Secondary endpoints included > or = 30% and > or = 50% reductions in NRS-3 scores, changes in neuropathic pain symptom inventory (NPSI) scores and allodynia severity ratings. Overall, 65.3% of patients treated with the 5% lidocaine medicated plaster and 62.0% receiving pregabalin responded to treatment with respect to the primary endpoint. A higher proportion of PHN patients responded to plaster treatment compared with pregabalin (63.0% vs 37.5%), whereas in the larger DPN group treatments were comparable. Both treatments improved NPSI scores and reduced allodynia severity. Patients administering lidocaine plaster experienced fewer drug-related adverse events (3.9% vs 39.2%) and there were substantially fewer discontinuations due to drug-related adverse events (1.3% vs 20.3%). CONCLUSION: After 4 weeks, 5% lidocaine medicated plaster treatment was associated with similar levels of analgesia in patients with PHN or DPN but substantially fewer frequent adverse events than pregabalin. PMID- 19301938 TI - Pharmacokinetics of anorectal nifedipine and lidocaine (lignocaine) ointment following haemorrhoidectomy: an open-label, single-dose, phase IV clinical study. AB - OBJECTIVES: This study aimed to assess whether topical anorectal application of an ointment containing nifedipine (0.3% w/w) and lidocaine (lignocaine) [1.5% w/w] to patients undergoing Milligan-Morgan haemorrhoidectomy achieves pharmacologically relevant serum concentrations of the active ingredients and has any haemodynamic effects or adverse effects. METHODS: A single dose of 3 g of study ointment was circumferentially applied inside the anus to 24 patients (17 males and 7 females) aged 23-71 years (mean +/- SD: 42.9 +/- 4.9 years) during postoperative dressing after Milligan-Morgan haemorrhoidectomy from March 2007 to January 2008. Blood samples for the determination of nifedipine and lidocaine serum concentrations were drawn before surgery and at 20, 40, 60, 90, 120, 240, 360, 480 and 720 minutes after application. Serum concentrations of nifedipine and lidocaine were determined by a high-performance liquid chromatography method in order to calculate pharmacokinetic parameters. Patients' BP, heart rate and ECG readings were monitored during the study. RESULTS: Chromatographic signals of nifedipine were sporadically observed in only five patients (20.8%), consistent with therapeutically negligible concentrations and insufficient to permit calculation of any pharmacokinetic parameters. The serum concentrations of nifedipine in these five patients ranged from 5.9 to 18.8 ng/mL. Lidocaine concentrations were detectable in all patients. The means +/- SD and medians of pharmacokinetic parameters for lidocaine were as follows: maximum serum concentration (C(max)) 245.1 +/- 370.8 ng/mL, 73.6 ng/mL; time to reach C(max) (t(max)) 69.2 +/- 78.3 minutes, 40 minutes; area under the serum concentration time curve from 0 to 6 hours (AUC(6)) 756.5 +/- 1254.1 ng.h/mL, 238.2 ng.h/mL. Only three patients had maximum serum concentrations above 1000 ng/mL (1037.8, 1044.75 and 1364.1 ng/mL). These outlier concentrations were four to five times lower than the threshold of CNS lidocaine toxicity (5000-6000 ng/mL). No serious local or systemic adverse events were observed throughout the study, and no subjects developed arrhythmias or significant ECG changes. Neither BP nor mean heart rate varied significantly after application of a single dose. CONCLUSIONS: This study demonstrates that single-dose topical application of an ointment containing nifedipine (0.3% w/w) and lidocaine (1.5% w/w) to patients undergoing Milligan-Morgan haemorrhoidectomy is safe to use. Following application onto damaged anorectal mucosa, nifedipine and lidocaine are absorbed into the bloodstream in small quantities that do not have any major implications for the safety of the product. Further studies are required to evaluate nifedipine and lidocaine concentrations in serum using a multiple-dose regimen. PMID- 19301939 TI - Relationship between decrease in ambulatory blood pressure and heart rate variability due to the effects of taking olmesartan medoxomil. AB - BACKGROUNDS AND METHODS: A higher degree of clinical efficacy with olmesartan compared with other angiotensin receptor blockers, has been reported by several sources. In this study of 31 examples of cases of essential hypertension, Holter electrocardiogram, ambulatory blood pressure monitoring and pulse wave velocity (PWV) measurements were performed before and after substituting olmesartan 20 mg for candesartan 8 mg antihypertensive drug therapy. RESULTS: Following the therapeutic change, daily average systolic and diastolic blood pressures were decreased by 6.7 +/- 9.3 mmHg and 3.6 +/- 8.3 mmHg, respectively, with olmesartan 20 mg; PWV was also significantly decreased. Holter electrocardiogram heart rate variability spectral analysis demonstrated that none of the very low frequency (VLF), high frequency (HF) and low frequency (LF)/HF components were significantly altered. However, a significant correlation was observed between the LF/HF component and blood pressure difference, when blood pressure and heart rate variability components in each individual case were studied. CONCLUSION: This study shows that olmesartan has a stronger antihypertensive effect in comparison to candesartan, and does not generate reflex sympathoexcitatory activity. PMID- 19301940 TI - Generic versus non-generic formulation of extended-release clarithromycin in patients with community-acquired respiratory tract infections: a prospective, randomized, comparative, investigator-blind, multicentre study. AB - BACKGROUND AND OBJECTIVE: There is a general concern about the use of multisource (generic) antibacterials in the clinical setting with registration based solely on bioequivalence data. In order to address this concern, two modified-release formulations of clarithromycin (i.e. the originator Klacid XL and the generic Klarithran MR) were compared in patients with acute community-acquired respiratory tract infections. METHODS: Patients presenting with tonsillopharyngitis, sinusitis or pneumonia were randomized to receive either of the test drugs provided they clinically qualified for empirical clarithromycin treatment. The study endpoints were clinical and bacteriological cure rates, tolerability and safety. The study was designed to test for non-inferiority with regard to cure rates. RESULTS: The main outcome of this study was that both agents had similar clinical (non-inferior) and bacteriological cure rates and demonstrated no difference in tolerability in patients. The study also demonstrated the clinical efficacy of clarithromycin when used as empirical treatment in patients with respiratory tract infections in community practice (i.e. 95% clinical cure rate). CONCLUSION: The clarithromycin extended-release multisource product (Klarithran MR) does not differ significantly from the originator (Klacid XL) and the clinical cure rate of the generic formulation is non-inferior to that of the originator. The two formulations are tolerated similarly. PMID- 19301941 TI - Pharmacokinetic interaction of ciprofloxacin with diclofenac: a single-dose, two period crossover study in healthy adult volunteers. AB - BACKGROUND AND OBJECTIVE: Ciprofloxacin is a broad-spectrum, synthetic antibacterial used for the treatment of various bacterial infections. In multidrug therapy, ciprofloxacin is commonly prescribed with analgesics for the management of infection, pain and inflammation. The objective of this study was to evaluate the pharmacokinetic properties of ciprofloxacin tablets with concurrent administration of diclofenac tablets in healthy adult human volunteers. METHODS AND DESIGN: The disposition pharmacokinetics of a single oral dose of ciprofloxacin 500 mg alone and with co-administration of a diclofenac 50 mg tablet in 12 healthy male volunteers was investigated using a two-period, crossover design. The blood samples were collected at 0 (predose), 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 16 and 24 hours after administration of the drugs and the concentration of ciprofloxacin in serum was determined using reversed phase high performance liquid chromatography. The pharmacokinetic parameters were calculated using a noncompartmental model and a two-compartment model. RESULTS: The maximum plasma concentration (C(max)) of ciprofloxacin increased from 2.48 +/- 0.33 microg/mL when administered alone to 3.91 +/- 0.8 microg/mL with co administration of diclofenac. Time to reach C(max) (t(max)) with ciprofloxacin reduced from 2.02 +/- 0.3 hours when administered alone to 1.49 +/- 0.2 h with co administration of diclofenac. Significant increases in ciprofloxacin area under the serum concentration-time curve (AUC) and elimination half-life, together with a significant decrease in total body clearance of ciprofloxacin, were observed with concurrent administration of diclofenac. CONCLUSION: Oral co-administration of ciprofloxacin tablets with diclofenac tablets increased ciprofloxacin AUC and C(max), and reduced ciprofloxacin t(max) and total body clearance. PMID- 19301942 TI - Effect of tributyltin on testicular development in Sebastiscus marmoratus and the mechanism involved. AB - Organotin compounds, such as tributyltin (TBT), that have been used as antifouling biocides can induce masculinization in female mollusks. However, few studies addressing the effects of TBT on fishes have been reported. The present study was conducted to investigate the effects of TBT at environmentally relevant concentrations (1, 10, and 100 ng/L) on testicular development in Sebastiscus marmoratus and to gain insight into its mechanism of action. After exposure for 48 d, the gonadosomatic index had decreased in a dose-dependent manner. Although the testosterone levels in the testes were elevated and the 17beta-estradiol levels were decreased, spermatogenesis was suppressed. Moreover, gamma-glutamyl transpeptidase activity (which is used as a Sertoli cell marker) was decreased in a dose-dependent manner after TBT exposure, and serious interstitial fibrosis was observed in the interlobular septa of the testes in the 100 ng/L TBT test group. Increases in the retinoid X receptors and peroxisome proliferator activated receptor gamma expression and the progressive enlargement of lipid droplets in the testes were observed after TBT exposure. Estrogen receptor alpha levels in the testes of the fish exposed to TBT decreased in a dose-dependent manner. The reduction of estrogen receptor alpha mRNA resulted from the decrease of 17beta estradiol levels, and the progressive enlargement of lipid droplets may have contributed to the dysfunction of the Sertoli cells, which then disrupted spermatogenesis. PMID- 19301943 TI - Toxicity of trace metals in soil as affected by soil type and aging after contamination: using calibrated bioavailability models to set ecological soil standards. AB - Total concentrations of metals in soil are poor predictors of toxicity. In the last decade, considerable effort has been made to demonstrate how metal toxicity is affected by the abiotic properties of soil. Here this information is collated and shows how these data have been used in the European Union for defining predicted-no-effect concentrations (PNECs) of Cd, Cu, Co, Ni, Pb, and Zn in soil. Bioavailability models have been calibrated using data from more than 500 new chronic toxicity tests in soils amended with soluble metal salts, in experimentally aged soils, and in field-contaminated soils. In general, soil pH was a good predictor of metal solubility but a poor predictor of metal toxicity across soils. Toxicity thresholds based on the free metal ion activity were generally more variable than those expressed on total soil metal, which can be explained, but not predicted, using the concept of the biotic ligand model. The toxicity thresholds based on total soil metal concentrations rise almost proportionally to the effective cation exchange capacity of soil. Total soil metal concentrations yielding 10% inhibition in freshly amended soils were up to 100-fold smaller (median 3.4-fold, n = 110 comparative tests) than those in corresponding aged soils or field-contaminated soils. The change in isotopically exchangeable metal in soil proved to be a conservative estimate of the change in toxicity upon aging. The PNEC values for specific soil types were calculated using this information. The corrections for aging and for modifying effects of soil properties in metal-salt-amended soils are shown to be the main factors by which PNEC values rise above the natural background range. PMID- 19301945 TI - Reduced expression of angiopoietin-1 in Hantaan virus-infected human umbilical vein endothelial cell increases their permeability. AB - Hemorrhagic fever with renal syndrome (HFRS) caused by Hantaan virus (HTNV) is characterized by vascular hemorrhage and acute renal failure. Angiopoietin-1 (Ang 1) is a glycoprotein that maintains vessel integrity and reduces endothelial permeability. We found that in HTNV-infected human umbilical vein endothelial cells (HUVECs) the levels of Ang-1 mRNA and protein were reduced on days 2 and 3 post-infection (p.i.), when endothelial permeability was increased. The HTNV stimulated permeability was reduced by treatment of HUVECs with Ang-1. The plasma Ang-1 level was lower in HFRS patients than in healthy persons. Paired plasma samples of HFRS patients revealed markedly lower Ang-1 levels during the acute phase of HFRS as compared to the convalescent phase. These findings suggested that HTNV reduced the Ang-1 expression in endothelial cells that might play an important role in the increase of vascular permeability in HFRS. PMID- 19301946 TI - Broadly cross-reactive monoclonal antibodies against HA2 glycopeptide of Influenza A virus hemagglutinin of H3 subtype reduce replication of influenza A viruses of human and avian origin. AB - The reactivity of monoclonal antibodies (MAbs) prepared to the HA2 glycopeptide (gp) of A/Dunedin/4/73 (H3N2) hemagglutinin was tested against influenza A viruses of H3, H4, and H7 subtypes. Only one (CF2) out of six MAbs reacted with influenza A viruses of all three subtypes (H3, H4 and H7). The inter-subtype reactivity of this MAb (CF2) is in accord with the highly conservative sequence in the previously defined MAb-binding site I, i.e. the aa 1-38 of N-terminus of HA2 gp. MAb CF2 as well as inter-subtype cross-reactive MAb IIF4, recognizing the binding site II of HA2 gp, were tested for their effect on replication of influenza A viruses. Both these MAbs reduced the number of plaques of viruses of homologous (H3) as well as heterologous (H4) virus subtypes, the latter less efficiently. The potential of these MAbs to influence in vivo replication of influenza A viruses of various subtypes is discussed. PMID- 19301947 TI - Development of a simple and effective method for specific detection of Pepper mild mottle virus. AB - A digoxigenin (DIG)-labeled cDNA probe complementary to the region from 5,256 6,300 nt of Pepper mild mottle virus (PMMoV) genome was synthesized. The specificity and sensitivity of the probe was tested by the dot-blot hybridization. The detection limit of this method was equivalent to 0.8 microg of fresh infected tissue in each spot. Double-antibody sandwich (DAS) ELISA and RT PCR had the detection limit 39 microg and 0.008 microg of fresh infected tissue, respectively. We evaluated leaf, fruit pulp, and seed of pepper plant by dot-blot hybridization and found all the tested tissues suitable for detection of PMMoV. Finally, 111 tissue samples including 93 samples collected from the pepper fields of Beijing and Baoding and 18 commercial seed samples were evaluated by this method. The results showed that the incidence rate of the infected samples was 14% and 61% for the field samples and commercial seeds, respectively. The high sensitivity and reliability of the molecular hybridization assay provided an important alternative method for the detection of PMMoV in a large-scale. PMID- 19301948 TI - Effects of disulfide bridges glycoprotein E1 on fusogenic activity of Rubella virus. AB - Rubella virus (RUBV) infects cells via an acid-triggered membrane fusion process. RUBV virions contain two cysteine-rich glycoproteins, E2 and E1. The latter is believed to be involved in the membrane fusion. Using a recombinant plasmid containing RUBV E1 and E2, 11 of total 20 cysteines present in the ectodomain of wild type E1 were mutated to test their role in the fusion via the formation of disulfide bridges. The recombinant plasmids containing mutated E1 (Cys2-Cys20) or wild type (wt) E1 were expressed in BHK-21 cells. Their fusogenic and hemadsorption activities in addition to a potential of cell surface expression of E1 and E2 were assayed. The results showed that the fusogenic activity was lost in all tested mutants, while the hemadsorption activity and cell surface expression potential were affected differently in individual mutants. Since only the Cys5 and Cys8 mutations led to a reduction of both hemadsorption and cell surface expression, we assume that these mutations prevented the formation of the disulfide bridge, what led to a misfolding of E1 and consequently to a failure of recognition of E1 by E2. In conclusion, the disulfide bridges disrupted in all the tested mutants appear essential for the cell fusion, while only the disulfide bridge C(5)-C(8) seems to be crucial for the transport of E1 and E2 in the cell. PMID- 19301949 TI - Immunogenic properties of recombinant ectodomain of Newcastle disease virus hemagglutinin-neuraminidase protein expressed in Escherichia coli. AB - Hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) plays a vital role in the viral infectivity, host immunity, and disease diagnosis. A portion of the HN gene encoding the ectodomain (nt 142-1739) was cloned and expressed in Escherichia coli yielding an insoluble HN protein and a soluble NusA HN protein containing N-utilization substance A (NusA) fusion component. Both recombinant proteins were purified and used for immunization of chickens. The recombinant HN protein induced higher antibody titers as compared to the recombinant NusA-HN protein. These antibodies were able to react in immunoblot analysis with the corresponding recombinant proteins as well as with the HN protein of NDV. PMID- 19301950 TI - Detection of Human papillomavirus and cellular regulators p16INK4a, p53, and NF kappaB in penile cancer cases in Kenya. AB - Human papillomavirus (HPV) E6 and E7 gene products play a central role in the induction of benign proliferation and malignant transformation by interacting with several cellular regulatory proteins such as p53, p16(INK4a), and nuclear factor kappaB (NF-kappaB). In this study, HPV DNA was detected by in situ hybridization (ISH) and p53, p16(INK4a), and NF-kappaB by immunochemistry in 22 penile cancer cases in Kenya. HPV DNA was found in 68.2% of the cases. There was no difference in the p53- and p16(INK4a)-positivities in HPV DNA-positive and HPV DNA-negative cases. In HPV DNA-positive cases, the NF-kappaB positivity in the nucleus, cytoplasm, and nucleus and/or cytoplasm amounted to 73.3%, 93.3%, and 100%, respectively, while in HPV DNA-negative cases, a 28.7% NF-kappaB positivity of in the nucleus and/or cytoplasm was observed. It is concluded that NF-kappaB in penile cancer is expressed more frequently in the presence of HPV infection than in its absence. PMID- 19301951 TI - Occurrence of Potato virus X on hybrid dock in Czech Republic. AB - Hybrid dock of Uteush (Rumex patientia L. x Rumex tianschanicus A. Los., the family Polygonaceae) is a perspective high productive crop and in the last decade its farming area has continuously grown in Czech Republic. However, the introduction of this non-native perennial crop into a present plant production creates a new potential reservoir for some plant viruses. Also, the hybrid dock could become a host of currently uncommon or insignificant viruses. We screened two dock-farming localities situated in south-west and north-east part of the Czech Republic for the presence of potyviruses, potexviruses, and carlaviruses. In the south-west part of the country, we detected a high incidence of Potato virus X (PVX, the genus Potexvirus). In contrast, in the north-east part of the country we did not detect any dock plants infected with PVX. Next, two other viruses, Turnip yellow mosaic virus (TYMV) and Radish mosaic virus (RaMV) were mechanically inoculated and tested for their survival capacity and multiplication in the hybrid dock. Both viruses were detected 9 months after inoculation in the infected plants. PMID- 19301952 TI - Differentiation of Plum pox virus isolates by single-strand conformation polymorphism and low-stringency single specific primer PCR analysis of HC-Pro genome region. AB - Single-strand conformation polymorphism (SSCP) and low-stringency single specific primer (LSSP)-PCR were assessed for suitability and reliability in genotyping of Plum pox virus (PPV) isolates. Examined PPV isolates included 16 PPV-D, 12 PPV-M, and 14 PPV-Rec isolates collected in Czech Republic. The analysis was performed on the helper component protease (HC-Pro) region of the PPV genome. SSCP and LSSP PCR allowed the differentiation of PPV strain, but SSCP was not able to distinguish isolates within the same strain. The individual genotyping of each PPV isolate was obtained by LSSP-PCR. Nevertheless, both SSCP and LSSP-PCR techniques are suitable for preliminary screening of genetic variability of plant RNA viruses. PMID- 19301953 TI - Anticancer effects of an oncolytic parvovirus combined with non-conventional therapeutics on pancreatic carcinoma cell lines. AB - Standard therapies such as surgery and chemotherapy offer only minimal improvement in pancreatic cancer. However, the viruses killing cancer cells and substances like some antibiotics and phytoalexins with anticancer potential may represent a candidate non-conventional mean of cancer treatment in the future. In this study, the effect of infection with oncolytic H-1 parvovirus (H-1PV) combined with antibiotic norfloxacin (NFX) or phytoalexin resveratrol on the survival of cell lines Panc-1 and BxPC3 derived from human pancreatic carcinoma was tested. Whereas H-1PV with NFX exerted a synergistic effect, H-1PV with resveratrol resulted in an additive effect only. All the effects were partial, but they were more pronounced in Panc-1 compared to BxPC3 cells. PMID- 19301954 TI - Molecular characterization of geographically different cucurbit aphid-borne yellows virus isolates. AB - Cucurbit aphid-borne yellows virus (CABYV) causes yellowing symptoms in cucurbit crops worldwide. In this work, the sequences for the coat protein (CP) gene for CABYV isolates from Iran and Slovakia are reported for the first time. Three Iranian isolates shared 96.2-99.3% and the Slovak isolate 98.9% of nucleotide identity in comparison with the reference French CABYV-N isolate. Phylogenetic analysis showed that the sequence of CP gene of examined CABYV isolates differed only slightly. Relatively close relationship and sequence similarity of geographically distant CABYV isolates could reflect the epidemic outbreak and rapid expansion of the virus throughout the world due to the host and vector abundance. PMID- 19301955 TI - Partial molecular characterization of an unusual, recombinant Plum pox virus isolate from Bulgaria. PMID- 19301956 TI - Notch-induced rat and human bone marrow stromal cell grafts reduce ischemic cell loss and ameliorate behavioral deficits in chronic stroke animals. AB - Gene transfection with Notch 1 intracellular domain and subsequent growth factor treatment stimulate neuron-like differentiation of bone marrow stromal cells (BMSCs). Here, we examined the potential of transplanting Notch-induced BMSCs to exert therapeutic effects in a rat model of chronic ischemic stroke. In experiment 1, Notch-induced rat BMSCs were intrastriatally transplanted in rats at 1 month after being subjected to transient occlusion of middle cerebral artery (MCAo). Compared to post-stroke/pretransplantation level, significant improvements in locomotor and neurological function were detected in stroke rats that received 100 k and 200 k BMSCs, but not in those that received 40 k BMSCs. Histological results revealed 9%-15% graft survival, which dose-dependently correlated with behavioral recovery. At 5 weeks post-transplantation, some grafted BMSCs were positive for the glial marker GFAP (about 5%), but only a few cells (2-5 cells per brain) were positive for the neuronal marker NeuN. However, at 12 weeks post-transplantation, where the number of GFAP-positive BMSCs was maintained (5%), there was a dramatic increase in NeuN-positive BMSCs (23%). In experiment 2, Notch-induced human BMSCs were intrastriatally transplanted in rats at 1 month following the same MCAo model. Improvements in both locomotor and neurological function were observed from day 7 to day 28 post-transplantation, with the high dose (180 k) displaying significantly better behavioral recovery than the low dose (90 k) or vehicle. There were no observable adverse behavioral effects during this study period that also involved chronic immunosuppression of all animals. Histological analyses revealed a modest 5%-7% graft survival, with few (<1%) cells expressing an intermediate MAP2 neuronal marker, but not glial or oligodendroglial markers. In addition, striatal peri-infarct cell loss was significantly reduced in transplanted stroke animals compared to vehicle-treated stroke animals. The present study demonstrates the potential of Notch-induced BMSC cell therapy for patients presenting with fixed ischemic stroke. PMID- 19301957 TI - Prospective comparison of quality of life before and after observation, radiation, or surgery for vestibular schwannomas. AB - OBJECT: The best management strategy for small-to medium-sized vestibular schwannomas (VSs) remains controversial between observation, radiation, and microsurgical resection (surgery). The authors undertook a prospective observational cohort study comparing all 3 therapeutic modalities in patients with VSs, focusing on quality of life (QOL) outcomes. METHODS: All patients in the study completed the 36-Item Short Form Health Survey at regular intervals, with a mean follow-up of 31.8 months. Two hundred five (77.7%) of 264 patients completed questionnaires, including 47 who underwent observation, 48 who received either linear accelerator radiosurgery or fractionated radiotherapy (radiation group), and 134 who underwent surgery (of whom 37 had tumors > 3 cm in diameter). Patients allocated to the observation group had smaller tumors than those in the other groups (mean 1.3 cm; p < 0.001). Patients who received radiation were older than patients in the other groups (mean 60.0 years; p < 0.001). RESULTS: There were no baseline QOL differences between the observation, radiation, and surgery (tumors 3 cm, there was an early improvement in composite mental dimension at 1.5 months, as well as at 24 months; total score and composite physical dimension were improved at 24 months in this group as well. CONCLUSIONS: Based on the current management protocol, patients with VSs enjoy similar QOL throughout the follow-up period after undergoing observation, radiation therapy, or surgery. PMID- 19301958 TI - Evaluation of new radiolucent polymer headholder pins for use in intraoperative computed tomography. AB - OBJECT: With the introduction of intraoperative CT (iCT) scanning, neurosurgeons can now obtain images of the brain during surgery, offering the possibility of intraoperative resection control and monitoring of potential intraoperative complications. The combination of iCT with neuronavigation makes it possible to update the reference scans intraoperatively when necessary. However, the headholder pins normally used for iCT scanning still show artifacts. In the present study, new polymer pins, producing nearly no artifacts in laboratory tests, are compared with the usual pins with regard to their mechanical and artifact behavior to evaluate their potential use in the clinical routine. METHODS: Pins made of different materials (titanium, Macor, silicon nitride, zirconium oxide, sapphire, polyetheretherketone, and polyparaphenylene copolymer) were used for the fixation of 10 cadaveric heads. Special force sensors measured the fixation pressure of the pins, and histological analysis revealed the penetration depth. Computed tomography scans of a head phantom, fixed with the different pins, were obtained to reveal artifact behavior. RESULTS: All pins were biocompatible. Pins did not differ significantly in fixation pressures and mechanical behavior. Penetration depths were comparable (maximum 1.4 mm) and did not cause opening of the diploe. Polymer pins made of polyparaphenylene showed the best results in artifact behavior in CT scans. CONCLUSIONS: The authors' results demonstrate that the new polymer pins are comparable in their mechanical behavior to the usual pins but superior in artifact behavior. Therefore, their use in the clinical routine of iCT scanning will be beneficial for the surgeon. PMID- 19301959 TI - Distal arterial occlusion for dissecting aneurysms. PMID- 19301960 TI - Vascularized temporoparietal fascial flap for the treatment of a traumatic cerebrospinal fluid fistula in the middle cranial fossa. Technical note. AB - The temporoparietal fascial flap (TPFF) is a vascularized pedicle flap commonly used in head and neck reconstructions. The authors describe the use of a TPFF in repairing a large traumatic middle cranial fossa CSF fistula via a middle cranial fossa approach. During the 18-month follow-up, the patient had no further CSF leakage or meningitis. The use of a TPFF for the repair of a traumatic CSF fistula has not been described in the literature. The authors recommend the use of a TPFF in treating recurrent or large middle cranial fossa CSF fistulas. The flap provides a well-vascularized thin layer that can be ideal in middle cranial fossa repair, and its proximity to the operation site is perfect as well. PMID- 19301961 TI - Avoiding the ventricle: a simple step to improve accuracy of anatomical targeting during deep brain stimulation. AB - OBJECT: The authors examined the accuracy of anatomical targeting during electrode implantation for deep brain stimulation in functional neurosurgical procedures. Special attention was focused on the impact that ventricular involvement of the electrode trajectory had on targeting accuracy. METHODS: The targeting error during electrode placement was assessed in 162 electrodes implanted in 109 patients at 2 centers. The targeting error was calculated as the shortest distance from the intended stereotactic coordinates to the final electrode trajectory as defined on postoperative stereotactic imaging. The trajectory of these electrodes in relation to the lateral ventricles was also analyzed on postoperative images. RESULTS: The trajectory of 68 electrodes involved the ventricle. The targeting error for all electrodes was calculated: the mean +/- SD and the 95% CI of the mean was 1.5 +/- 1.0 and 0.1 mm, respectively. The same calculations for targeting error for electrode trajectories that did not involve the ventricle were 1.2 +/- 0.7 and 0.1 mm. A significantly larger targeting error was seen in trajectories that involved the ventricle (1.9 +/- 1.1 and 0.3 mm; p < 0.001). Thirty electrodes (19%) required multiple passes before final electrode implantation on the basis of physiological and/or clinical observations. There was a significant association between an increased requirement for multiple brain passes and ventricular involvement in the trajectory (p < 0.01). CONCLUSIONS: Planning an electrode trajectory that avoids the ventricles is a simple precaution that significantly improves the accuracy of anatomical targeting during electrode placement for deep brain stimulation. Avoidance of the ventricles appears to reduce the need for multiple passes through the brain to reach the desired target as defined by clinical and physiological observations. PMID- 19301962 TI - Analysis of cerebral perfusion and metabolism assessed with positron emission tomography before and after carotid artery stenting. Clinical article. AB - OBJECT: The authors analyzed cerebral perfusion and metabolism in patients with internal carotid artery stenosis before and after carotid artery stenting (CAS). METHODS: Sixteen patients with internal carotid artery stenosis (>70%) underwent PET scanning before CAS, 1-7 days after CAS, and 3-4 months after CAS to assess a variety of parameters related to cerebral perfusion and metabolism. RESULTS: Cerebral blood flow at rest (CBFrest) significantly increased in the immediate postoperative stage before returning to normal levels over the long term; this trend was also recognized on the contralateral side. In contrast, there was gradual improvement in the rate of CBF variation on acetazolamide administration (% CBFaz). Cerebral perfusion pressure (CBF/cerebral blood volume) increased rapidly during the acute stage and decreased in the long term, and the oxygen extraction fraction decreased slightly during the acute stage before normalizing over the long term. The cerebral metabolic rate of oxygen (CMRO2) increased slightly after stenting over both the short and long term. The ratios of ipsilateral to contralateral values (asymmetry index) for CBFrest, % CBFaz, cerebral blood volume, oxygen extraction fraction, and CMRO2 tended to approach 1.0 over time. CONCLUSIONS: Repeated PET scanning revealed improvements in CBF, perfusion pressure, and oxygen metabolism after CAS. In particular, the vascular reserve tended to improve gradually, while CBF, cerebral perfusion pressure, and CMRO2 increased rapidly and peaked soon after CAS. These results suggest that a large discrepancy between rapidly increased CBF, perfusion pressure, and a small increase in vascular reserve in the acute stage after CAS could cause hyperperfusion syndrome. PMID- 19301963 TI - Magnetic resonance imaging for preoperative identification of the lenticulostriate arteries in insular glioma surgery. Technical note. AB - Aggressive resection of insular tumors is possible using a meticulous surgical approach based on the regional insular anatomy, with high rates of gross-total resection and low rates of permanent neurological deficits. However, the risk of postoperative morbidities remains high, often caused by disruption of the vascular supply to the surrounding eloquent structures, especially the lenticulostriate arteries (LSAs) that supply the internal capsule. Three dimensional 3-T time-of-flight (TOF) MR imaging was performed pre- and postoperatively in patients with insuloopercular gliomas. This 3D 3-T TOF MR imaging clearly visualized the LSAs and the relationships with the tumor margins. These findings were confirmed intraoperatively. Three-dimensional 3-T TOF MR imaging of the LSAs in patients with insuloopercular gliomas can help to maximize the extent of resection without neurological complications, preserve the LSAs during surgery, and assist in patient selection. PMID- 19301964 TI - Remote distal outflow occlusion: a novel treatment option for complex dissecting aneurysms of the posterior inferior cerebellar artery. Report of 3 cases. AB - The authors report a novel management option for patients with complex dissecting aneurysms of the posterior inferior cerebellar artery (PICA). The authors reviewed the medical records and neuroimaging studies of 3 patients who underwent a novel surgical treatment for complicated dissecting PICA aneurysms. The mean follow-up period was 1.1 years, and no patient was lost to follow-up. Two patients were in poor condition following an acute, severe subarachnoid hemorrhage, and 1 presented with headaches and a remote history of bleeding. All patients underwent surgical occlusion of the PICA beyond the tonsillar loop, distal to the aneurysmal segment. Intraoperative and delayed follow-up angiography demonstrated progressive diminution in size of the aneurysmal dilation but persistent filling of the proximal PICA segments supplying the brainstem. Outcome was good in all cases. This novel technique has been used successfully in 3 cases and, to the authors' knowledge, has not been reported previously. PMID- 19301965 TI - Flow dynamics in lumboperitoneal shunts and their implications in vivo. AB - OBJECT: Lumboperitoneal shunting is the standard treatment for pseudotumour cerebri or idiopathic intracranial hypertension. Complications are common, particularly the problem of overdrainage leading to low pressure symptoms. The authors designed a simple experiment using catheters of different lengths that drained at different pressure heads and with different vertical drops to study the flow characteristics in these shunts and determine the optimal catheter placement and length that would reduce the occurrence of low pressure headaches. METHODS: The flow rates through catheters of 3 different lengths (60, 83, and 100 cm) with the same internal radius, at 3 different pressure heads (15, 25, and 35 cm H2O to simulate 3 different placements in the lumbar theca), and 3 different vertical drops (10, 20, and 30 cm to simulate the possible effect of siphoning) were measured and the results analyzed. RESULTS: Application of Poiseuille's law and Bernoulli's principle to the experimental design shows that the volume of flow is directly proportional to the sum of the pressure head and the vertical drop and inversely proportional to the length of the catheter. The flow rate through the standard catheter lengths over the course of 24 hours can be abnormally high. An attempt to predict the optimal catheter length was made. CONCLUSIONS: Although the catheter position in the theca and abdomen cannot be altered significantly and the internal radius of the tube cannot be reduced further without increasing the risk of blockage, the length of the tube can be increased to combat overdrainage. The authors suggest that currently available catheters are too short. PMID- 19301966 TI - Editorial. Mesenchymal stem cell treatment of traumatic brain injury. PMID- 19301967 TI - Candidate genes for sensitivity and resistance of human glioblastoma multiforme cell lines to erlotinib. Laboratory investigation. AB - OBJECT: The authors have previously reported that erlotinib, an EGFR tyrosine kinase inhibitor, exerts widely variable antiproliferative effects on 9 human glioblastoma multiforme (GBM) cell lines in vitro and in vivo. These effects were independent of EGFR baseline expression levels, raising the possibility that more complex genetic properties form the molecular basis of the erlotinib-sensitive and erlotinib-resistant GBM phenotypes. The aim of the present study was to determine candidate genes for mediating the cellular response of human GBMs to erlotinib. METHODS: Complementary RNA obtained in cell lines selected to represent the sensitive, somewhat responsive, and resistant phenotypes were hybridized to CodeLink Human Whole Genome Bioarrays. RESULTS: Expression analysis of 814 prospectively selected genes involved in major proliferation and apoptosis signaling pathways identified 19 genes whose expression significantly correlated with phenotype. Functional annotation analysis revealed that 2 genes (DUSP4 and STAT1) were significantly associated with sensitivity to erlotinib, and 10 genes (CACNG4, FGFR4, HSPA1B, HSPB1, NFATC1, NTRK1, RAC1, SMO, TCF7L1, and TGFB3) were associated with resistance to erlotinib. Moreover, 5 genes (BDNF, CARD6, FOSL1, HSPA9B, and MYC) involved in antiapoptotic pathways were unexpectedly found to be associated with sensitivity. Gene expressions were confirmed by quantitative polymerase chain reaction. CONCLUSIONS: Based on an analysis of gene expressions in cell lines with sensitive, somewhat responsive, and resistant phenotypes, the authors propose candidate genes for GBM response to erlotinib. The 10 gene candidates for conferring GBM resistance to erlotinib may represent therapeutic targets for enhancing the efficacy of erlotinib against GBMs. Five additional genes warrant further investigation into their role as putative cotargets of erlotinib. PMID- 19301968 TI - Stereotactic radiosurgery for metastatic brain tumors: a comprehensive review of complications. AB - OBJECT: Stereotactic radiosurgery (SRS) is commonly used to treat brain metastases. Complications associated with this treatment are underreported. The authors reviewed a large series of patients who underwent SRS for brain metastases to identify complications and factors predicting their occurrence. METHODS: Prospectively collected clinical data from 273 patients undergoing SRS for 1 or 2 brain metastases at The University of Texas M. D. Anderson Cancer Center between June 1993 and December 2004 were reviewed. Patients who had received prior treatment for their tumor, including whole-brain radiation, SRS, or surgery, were excluded from the study. Data on adverse neurological and nonneurological outcomes following treatment were collected. RESULTS: Three hundred sixteen lesions were treated. Complications were associated with 127 (40%) of 316 treated lesions. New neurological complications were associated with 101 (32%) of 316 lesions. The onset of seizure was the most common complication, occurring in 41 (13%) of 316 SRS cases. On multivariate analysis, progressing primary cancer (hazard ratio [HR]=2.4, 95% CI 1.6-3.6, p<0.001), tumor location in eloquent cortex (HR=2.3, 95% CI 1.6-3.4, p<0.001), and lower (<15 Gy) SRS dose (HR=2.1, 95% CI 1.1-4.2, p=0.04) were significantly associated with new complications. On multivariate analysis, a tumor location in the eloquent cortex (HR=2.5, 95% CI 1.6-3.8, p<0.001) and progressing primary cancer (HR=1.6, 95% CI 1.1-2.5, p=0.03) were significantly associated with new neurological complications. CONCLUSIONS: The authors showed that new neurological and nonneurological complications were associated with 40% of SRS treatments for brain metastases. Patients with lesions in functional brain regions have a significantly increased risk of treatment-related complications. PMID- 19301969 TI - Predictors and outcome of seizures after spontaneous intracerebral hemorrhage. Clinical article. AB - OBJECT: Seizures are an important neurological complication of spontaneous intracerebral hemorrhage (ICH). A better understanding of the risk factors of seizures following ICH is needed to predict which patients will require treatment. METHODS: Two hundred and forty-three adult patients were enrolled in this 1-year retrospective study. Multiple logistic regression was used to evaluate the relationship between baseline clinical factors and the presence or absence of seizure during the study period. RESULTS: Seizures occurred in 20 patients with ICH, including acute symptomatic seizures in 9 and unprovoked seizures in 11. None progressed to status epilepticus during hospitalization. After a minimum 3-year follow-up period, the mean Glasgow Outcome Scale score was 3.8+/-1.1 for patients who had had seizures and 3.5+/-1.3 for those who had not. The multiple logistic regression model demonstrated that the mean ICH volume was independently associated with seizures, and any increase of 1 mm3 in ICH volume increased the seizure rate by 2.7%. CONCLUSIONS: Higher mean ICH volumes at presentation were predictive of seizure, and the presence of late seizures was predictive of developing epilepsy. Most seizures occurred within 2 years of spontaneous ICH over a minimum of 3 years of follow-up. PMID- 19301970 TI - Combined transarterial and percutaneous coiling of a spontaneous vertebrovertebral fistula associated with neurofibromatosis Type 1. Case report. AB - The authors report a rare case of spontaneous left vertebrovertebral fistula associated with neurofibromatosis Type 1 in a young woman. The patient had undergone previous failed surgical attempts to close the fistula and presented to the authors with recurrence of the fistula and neurological deterioration. Successful obliteration of the fistula with combined transarterial and percutaneous coiling with Guglielmi detachable coils resulted in disappearance of the fistula and significant neurological improvement. PMID- 19301971 TI - Intraoperative biplanar rotational angiography during neurovascular surgery. Technical note. AB - OBJECT: The purpose of this study was to evaluate the authors' initial experience with the integration of high-resolution rotational and biplanar angiography during neurovascular operative procedures. METHODS: Eight patients with intracerebral arteriovenous malformations (AVMs) and aneurysms underwent surgical treatment of their lesions in a combined endovascular surgical suite. After initial head positioning, preoperative biplane and rotational angiography was performed. Resection of the AVM or clipping of the aneurysm was then performed. Further biplane and rotational 3D angiograms were obtained intraoperatively to confirm satisfactory treatment. RESULTS: One small residual AVM identified intraoperatively necessitated further resection. One aneurysm was clipped during endovascular inflation of an intracarotid balloon for temporary proximal control. The completeness of treatment was confirmed on intraoperative 3D rotational angiography in all cases, and there were no procedure-related complications. CONCLUSIONS: Intraoperative rotational angiography performed in an integrated biplane angiography/surgery suite is a safe and useful adjunct to surgery and may enable combining endovascular and surgical procedures for the treatment of complex vascular lesions. PMID- 19301972 TI - Assessing the microlesion effect of subthalamic deep brain stimulation surgery with FDG PET. AB - OBJECT: The authors investigated whether the insertion of deep brain stimulation electrodes into the subthalamic nucleus can alter regional brain metabolism in the absence of stimulation. METHODS: Six patients with Parkinson disease (PD) underwent preoperative FDG PET scanning, and again after STN electrode implantation with stimulation turned off. RESULTS: Compared with baseline values, glucose utilization was reduced in the postoperative off-stimulation scans in the putamen/globus pallidus and in the ventral thalamus (p < 0.01), and there was increased metabolism in the sensorimotor cortex and cerebellum (p < 0.005). The expression of a specific PD-related spatial covariance pattern measured in the FDG PET data did not change after electrode implantation (p = 0.36), nor was there a significant change in clinical motor ratings (p = 0.44). Differences in PD-related spatial covariance pattern expression among the patients after electrode implantation did, however, correlate with the number of microelectrode recording trajectories placed during surgery (r = -0.82, p < 0.05). CONCLUSIONS: These findings suggest that electrode implantation can impart a microlesion effect on regional brain function. Nonetheless, these local changes did not cross the threshold of network modulation needed to achieve clinical benefit. PMID- 19301973 TI - Intravenous mesenchymal stem cell therapy for traumatic brain injury. AB - OBJECT: Cell therapy has shown preclinical promise in the treatment of many diseases, and its application is being translated to the clinical arena. Intravenous mesenchymal stem cell (MSC) therapy has been shown to improve functional recovery after traumatic brain injury (TBI). Herein, the authors report on their attempts to reproduce such observations, including detailed characterizations of the MSC population, non-bromodeoxyuridine-based cell labeling, macroscopic and microscopic cell tracking, quantification of cells traversing the pulmonary microvasculature, and well-validated measurement of motor and cognitive function recovery. METHODS: Rat MSCs were isolated, expanded in vitro, immunophenotyped, and labeled. Four million MSCs were intravenously infused into Sprague-Dawley rats 24 hours after receiving a moderate, unilateral controlled cortical impact TBI. Infrared macroscopic cell tracking was used to identify cell distribution. Immunohistochemical analysis of brain and lung tissues 48 hours and 2 weeks postinfusion revealed transplanted cells in these locations, and these cells were quantified. Intraarterial blood sampling and flow cytometry were used to quantify the number of transplanted cells reaching the arterial circulation. Motor and cognitive behavioral testing was performed to evaluate functional recovery. RESULTS: At 48 hours post-MSC infusion, the majority of cells were localized to the lungs. Between 1.5 and 3.7% of the infused cells were estimated to traverse the lungs and reach the arterial circulation, 0.295% reached the carotid artery, and a very small percentage reached the cerebral parenchyma (0.0005%) and remained there. Almost no cells were identified in the brain tissue at 2 weeks postinfusion. No motor or cognitive functional improvements in recovery were identified. CONCLUSIONS: The intravenous infusion of MSCs appeared neither to result in significant acute or prolonged cerebral engraftment of cells nor to modify the recovery of motor or cognitive function. Less than 4% of the infused cells were likely to traverse the pulmonary microvasculature and reach the arterial circulation, a phenomenon termed the "pulmonary first-pass effect," which may limit the efficacy of this therapeutic approach. The data in this study contradict the findings of previous reports and highlight the potential shortcomings of acute, single-dose, intravenous MSC therapy for TBI. PMID- 19301974 TI - Methylation regulates hepatitis B viral protein expression. AB - BACKGROUND: Hepatitis B virus (HBV) DNA has been shown to contain CpG islands that are methylated in human tissue, which suggests a role for methylation in regulating viral protein production. However, data are lacking about whether methylation regulates viral gene expression. METHODS: To investigate the hypothesis that methylation of viral DNA regulates viral gene expression, unmethylated, partially methylated, and fully methylated viral DNA was transfected into HepG2 cells. In addition, a new assay was designed that specifically identifies methylated covalently closed circular DNA (cccDNA) in human liver tissue. RESULTS: Transfection of methylated HBV DNA led to reduced HBV mRNA levels in HepG2 cells, decreased surface and core protein expression in these cells, and decreased secretion of HBV viral proteins into the cell supernatant. These data provide direct evidence that CpG islands regulate gene transcription of HBV. Furthermore, methylated cccDNA was found in tumor and nonneoplastic human liver tissues. Finally, an in vitro equivalent of cccDNA showed decreased viral protein production in HepG2 cells after DNA methylation. CONCLUSION: Taken together, these data demonstrate that methylation of viral CpG islands can regulate viral protein production. PMID- 19301975 TI - A common CD4 gene variant is associated with an increased risk of HIV-1 infection in Kenyan female commercial sex workers. AB - BACKGROUND: It has been predicted that CD4 C868T, a novel CD4 single-nucleotide polymorphism (SNP) that has been found to be highly prevalent among Africans, changes the tertiary structure of CD4, which may alter susceptibility to human immunodeficiency virus (HIV) infection. METHODS: Participants were from a Kenyan cohort and included 87 uninfected and 277 HIV-1-infected individuals. DNA sequencing was used to determine CD4 genotype. A2.01 cells expressing similar levels of either wild-type CD4 or CD4-Trp240 as well as peripheral blood mononuclear cells from uninfected donors were infected with HIV-1(IIIB) or a Kenyan primary HIV-1 isolate. HIV-1 p24 enzyme-linked immunosorbent assay was used to determine the outcome of infection. RESULTS: CD4 C868T was found to be significantly more prevalent among HIV-1-infected participants than among HIV-1 uninfected participants (P = .002), and C868T was associated with an increased incidence of HIV-1 infection as well (P = .005, log-rank test; P = .009, Wilcoxon test), with an odds ratio of 2.49 (P = .009). Both in vitro and ex vivo models demonstrated a significant association between CD4 C868T and susceptibility to HIV-1 infection (P < .001 and P = .003, respectively). CONCLUSION: Overall, the present study found a strong correlation between CD4 C868T and increased susceptibility to HIV-1 infection. Given the high prevalence of both HIV infection and CD4 C868T in African populations, the effect of this SNP on the epidemic in Africa could be dramatic. PMID- 19301976 TI - Ultra-deep pyrosequencing of hepatitis B virus quasispecies from nucleoside and nucleotide reverse-transcriptase inhibitor (NRTI)-treated patients and NRTI-naive patients. AB - The dynamics of emerging nucleoside and nucleotide reverse-transcriptase inhibitor (NRTI) resistance in hepatitis B virus (HBV) are not well understood because standard dideoxynucleotide direct polymerase chain reaction (PCR) sequencing assays detect drug-resistance mutations only after they have become dominant. To obtain insight into NRTI resistance, we used a new sequencing technology to characterize the spectrum of low-prevalence NRTI-resistance mutations in HBV obtained from 20 plasma samples from 11 NRTI-treated patients and 17 plasma samples from 17 NRTI-naive patients, by using standard direct PCR sequencing and ultra-deep pyrosequencing (UDPS). UDPS detected drug-resistance mutations that were not detected by PCR in 10 samples from 5 NRTI-treated patients, including the lamivudine-resistance mutation V173L (in 5 samples), the entecavir-resistance mutations T184S (in 2 samples) and S202G (in 1 sample), the adefovir-resistance mutation N236T (in 1 sample), and the lamivudine and adefovir resistance mutations V173L, L180M, A181T, and M204V (in 1 sample). G-to-A hypermutation mediated by the apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like family of cytidine deaminases was estimated to be present in 0.6% of reverse-transcriptase genes. Genotype A coinfection was detected by UDPS in each of 3 patients in whom genotype G virus was detected by direct PCR sequencing. UDPS detected low-prevalence HBV variants with NRTI-resistance mutations, G-to-A hypermutation, and low-level dual genotype infection with a sensitivity not previously possible. PMID- 19301977 TI - Hydrogen peroxide-producing lactobacilli inhibit gonococci in vitro but not during experimental genital tract infection. AB - Commensal lactobacilli that produce hydrogen peroxide (H(2)O(2)) inhibit Neisseria gonorrhoeae in vitro, and clinical data suggest that they are associated with a reduced risk of gonorrhea. We precolonized mice with Lactobacillus crispatus and then challenged them with N. gonorrhoeae, to measure the effects of H(2)O(2)-producing lactobacilli on gonococcal infection. We found no difference in the duration of infection or the number of gonococci recovered from untreated mice and mice colonized with L. crispatus. A gonococcal catalase mutant and a catalase, cytochrome C peroxidase mutant exhibited greater susceptibility to L. crispatus in vitro than did wild-type bacteria; however, recovery of these mutants from mice was not affected by L. crispatus. We also found no evidence that utilization of lactobacillus-produced lactate by N. gonorrhoeae balances the detrimental effects of H(2)O(2) during infection. We conclude that the association between lactobacilli and gonococci is complex and may be subject to factors that have not been reproduced in vitro. PMID- 19301978 TI - Anogenital hairs are an important reservoir of alpha-papillomaviruses in patients with genital warts. AB - Human papillomaviruses (HPVs) were detected in 69 (43.7%) of 158 and in 7 (4.5%) of 155 anogenital hairs obtained from 53 patients with genital warts (GWs) and from 53 age-matched healthy control subjects, respectively. At least 1 hair sample was positive for 69.8% of patients and for 13.2% of control subjects. For patients, HPV was detected in 64.2%, 39.6%, and 26.9% of hairs plucked from the pubic, scrotal, and perianal regions, respectively. For 91.9% of patients, the same HPV genotype was identified in GWs and hairs from at least 1 sampling site. Having GWs was found to be strongly associated with the presence in anogenital hairs of the HPV genotype causing the GWs (range of odds ratios, 13.0-20.0). PMID- 19301979 TI - Long-term serologic responses to the Pneumocystis jirovecii major surface glycoprotein in HIV-positive individuals with and without P. jirovecii infection. AB - BACKGROUND: The immune responses to Pneumocystis jirovecii major surface glycoprotein (Msg) in individuals with human immunodeficiency virus (HIV) infection are poorly understood. METHODS: We examined the sequential serologic responses to recombinant Msg carboxyl terminus fragments (MsgC1, MsgC3, MsgC8, and MsgC9) by enzyme-linked immunosorbent assay in a cohort of individuals with HIV infection for the 5.5 years before death and autopsy. Analyses included mean antibody levels by status at death (Pneumocystis pneumonia, P. jirovecii colonization, or neither), factors associated with high antibody levels, and antibody responses before and after active Pneumocystis pneumonia. RESULTS: Patients who died from Pneumocystis pneumonia had higher levels of antibody to MsgC8 than did patients who died from other causes. Previous episode of Pneumocystis pneumonia, geographic location, and age were independent predictors of high levels of anitbodies to most or all Msgs. Failure to take Pneumocystis pneumonia prophylaxis was associated with high levels of antibody to MsgC1. Patients who developed and recovered from active Pneumocystis pneumonia during the study exhibited an increase in serum antibody levels that persisted for months after the infection, whereas patients who developed another acquired immunodeficiency syndrome-defining illness did not. CONCLUSIONS: Serum antibodies to Msgs are important markers of P. jirovecii infection in patients with HIV infection and are influenced by host and environmental factors in complex ways. PMID- 19301980 TI - Decreased immune activation in resistance to HIV-1 infection is associated with an elevated frequency of CD4(+)CD25(+)FOXP3(+) regulatory T cells. AB - Human immunodeficiency virus (HIV)-resistant commercial sex workers provide a unique opportunity to study correlates of protection associated with natural resistance to HIV infection. Emerging data from studies of these individuals and other uninfected individuals who have been exposed to HIV suggest that low levels of immune activation may contribute to protection against infection. In the present study, HIV-resistant individuals were shown to have reduced frequencies of T cells expressing the activation marker CD69. They were also found to have elevated frequencies of regulatory T (T(reg)) cells, compared with HIV-negative control individuals. By controlling levels of T cell activation, T(reg) cells may contribute to HIV resistance by minimizing the pool of cells susceptible to infection. PMID- 19301982 TI - Circulating HIV type 1 drug resistance will have limited impact on the effectiveness of preexposure prophylaxis among young women in Zimbabwe. AB - BACKGROUND: Preexposure prophylaxis (PrEP) with antiretroviral drugs may prevent transmission of human immunodeficiency virus (HIV). Our objective was to predict whether PrEP, in the presence of circulating drug resistance, will reduce the risk of infection with HIV. METHODS: We used risk equations to calculate the monthly risk of infection with HIV before and after the introduction of PrEP. Uncertainty and sensitivity analyses were performed for 2 ranges of PrEP effectiveness (40%-60% and 60%-80%). Circulating drug resistance was assumed to reduce the effectiveness of PrEP by 50%-90% and the transmissibility of HIV by 0% 30%. Parameter ranges were chosen for women 17-29 years of age from publications on HIV in Manicaland in Zimbabwe. RESULTS: PrEP would decrease the median risk of HIV transmission by 21%-33% (effectiveness of PrEP, 40%-60% and 60%-80%). If 50% of HIV strains are drug resistant, then the median risk reduction would be 19% 26% if drug-resistant strains were less transmissible than wild-type HIV and 12% 19% if they were equally transmissible. The risk would increase if condoms were frequently replaced with PrEP. Use of PrEP for sexual acts for which no protection is currently used would be beneficial. CONCLUSION: The public health impact of PrEP will depend on its effectiveness and on risk behavior. Circulating drug resistance will have only a small impact on the effectiveness of PrEP. PMID- 19301981 TI - The antibiotics doxycycline and minocycline inhibit the inflammatory responses to the Lyme disease spirochete Borrelia burgdorferi. AB - Tetracyclines moderate inflammatory responses of various etiologies. We hypothesized that tetracyclines, in addition to their antimicrobial function, could exert control over the inflammation elicited by Borrelia burgdorferi. To model systemic effects, we used the human monocytic cell line THP-1; to model effects in the central nervous system, we used rhesus monkey brain astrocytes and microglia. Cells were stimulated with live or sonicated B. burgdorferi or with the lipoprotein outer surface protein A in the presence of increasing concentrations of doxycycline or minocycline. Both antibiotics significantly reduced the production of tumor necrosis factor-alpha, interleukin (IL)-6, and IL 8 in a dose-dependent manner in all cell types. Microarray analyses of the effect of doxycycline on gene transcription in spirochete-stimulated monocytes revealed that the NFKB and CHUK (alias, IKKA) genes were down-regulated. Functionally, phosphorylation of IkappaBalpha and binding of NF-kappaB to target DNA were both reduced in these cells. Our results suggest that tetracyclines may have a dual therapeutic effect in Lyme disease. PMID- 19301983 TI - Accounting for incomplete postdischarge follow-up during surveillance of surgical site infection by use of the National Nosocomial Infections Surveillance system's risk index. AB - OBJECTIVE: We examined the usefulness of a simple method to account for incomplete postdischarge follow-up during surveillance of surgical site infection (SSI) by use of the National Nosocomial Infections Surveillance (NNIS) system's risk index. DESIGN: Retrospective cohort study that used data prospectively collected from 1993 through 2006. SETTING: Five private, nonuniversity healthcare facilities in Belo Horizonte, Brazil. PATIENTS: Consecutive patients undergoing the following NNIS operative procedures: 20,981 operations on the genitourinary system, 11,930 abdominal hysterectomies, 7,696 herniorraphies, 6,002 cholecystectomies, and 6,892 laparotomies. METHODS: For each operative procedure category, 2 SSI risk models were specified. First, a model based on the NNIS system's risk index variables was specified (hereafter referred to as the NNIS based model). Second, a modified model (hereafter referred to as the modified NNIS-based model), which was also based on the NNIS system's risk index, was specified with a postdischarge surveillance indicator, which was assigned the value of 1 if the patient could be reached during follow-up and a value of 0 if the patient could not be reached. A formal comparison of the capabilities of the 2 models to assess the risk of SSI was conducted using measures of calibration (by use of the Pearson goodness-of-fit test) and discrimination (by use of receiver operating characteristic curves). Goodman-Kruskal correlations (G) were also calculated. RESULTS: The rate of incomplete postdischarge follow-up varied between 29.8% for abdominal hysterectomies and 50.5% for cholecystectomies. The modified NNIS-based model for laparotomy did not show any significant benefit over the NNIS-based model in any measure. For all other operative procedures, the modified NNIS-based model showed a significantly improved discriminatory ability and higher G statistics, compared with the NNIS-based model, with no significant impairment in calibration, except if used to assess the risk of SSI after operations on the genitourinary system or after a cholecystectomy. CONCLUSIONS: Compared with the NNIS-based model, the modified NNIS-based model added potentially useful clinical information regarding most of the operative procedures. Further work is warranted to evaluate this method for accounting for incomplete postdischarge follow-up during surveillance of SSI. PMID- 19301984 TI - Vancomycin use during left ventricular assist device support. AB - We reviewed the frequency and duration of vancomycin use during 93 left ventricular assist device placements. Vancomycin prophylaxis was administered for a mean duration (+/- standard deviation) of 10.5 +/- 11 days. Empirical vancomycin use was frequent, with a mean duration (+/- standard deviation) of therapy of 9.8 +/- 9 days (median, 8 days) given during 81 (87%) of the implantations. The most common indications for empirical vancomycin treatment were isolated leukocytosis or driveline drainage. Strategies to improve vancomycin use during left ventricular assist device support should be considered. PMID- 19301985 TI - Success of an infection control program to reduce the spread of carbapenem resistant Klebsiella pneumoniae. AB - OBJECTIVE: To assess the effect of enhanced infection control measures with screening for gastrointestinal colonization on limiting the spread of carbapenem resistant Klebsiella pneumoniae in a New York City hospital endemic for this pathogen. DESIGN: Retrospective observational study with pre- and postinterventional phases. METHODS: Beginning in 2006, a comprehensive infection control program was instituted in a 10-bed medical and surgical intensive care unit at a university-based medical center. In addition to being placed in contact isolation, all patients colonized or infected with carbapenem-resistant gram negative bacilli, vancomycin-resistant Enterococcus, or methicillin-resistant Staphylococcus aureus were cohorted to one end of the unit. Improved decontamination of hands and environmental surfaces was encouraged. In addition, routine rectal surveillance cultures were screened for the presence of carbapenem resistant pathogens. The number of patients per quarter with clinical cultures positive for carbapenem-resistant K. pneumoniae was compared during the approximately 2-year periods before and after the intervention. RESULTS: The mean number (+/-SD) of new patients per 1,000 patient-days per quarter with cultures yielding carbapenem-resistant K. pneumoniae decreased from 9.7 +/- 2.2 before the intervention to 3.7 +/- 1.6 after the intervention (P < .001). There was no change in the mean number of patient-days or the mean number of patients per quarter with cultures yielding methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus, or carbapenem-resistant Acinetobacter baumannii or Pseudomonas aeruginosa after the intervention. There was no association between antibiotic usage patterns and carbapenem-resistant K. pneumoniae. CONCLUSIONS: The comprehensive intervention that combined intensified infection control measures with routine rectal surveillance cultures was helpful in reducing the incidence of carbapenem-resistant K. pneumoniae in an intensive care unit where strains producing the carbapenemase KPC were endemic. PMID- 19301986 TI - Breastmilk from allergic mothers can protect offspring from allergic airway inflammation. AB - OBJECTIVE: Breastfeeding is associated with a reduced risk of developing asthma in children. Using a murine model we previously demonstrated that mothers with Th1-type immunity to ovalbumin (OVA) transfer antigen-specific protection from OVA-induced allergic airway disease (AAD) to their offspring. The aim of this study was to evaluate the contribution of breastmilk and maternal B cell immunity from allergic mothers in the vertical transmission of protection from AAD. METHODS: This was investigated using an adoptive nursing strategy. Naive offspring were nursed by allergic wild-type or B cell-deficient foster mothers with histories of Th2-type immunity to OVA. Following weaning, offspring were immunized with OVA-Al(OH)(3) and challenged with aerosolized OVA to induce AAD. RESULTS: Offspring nursed by wild-type OVA-immune foster mothers demonstrated lower levels of OVA-specific immunoglobulin E, interleukin-5, and airway eosinophilia than progeny nursed by naive control mothers. In contrast, offspring nursed by B cell-deficient OVA-immune foster mothers had similar parameters of OVA-induced AAD as progeny nursed by naive control mothers. CONCLUSIONS: These data demonstrate the ability of breastmilk from allergic mothers to protect offspring from AAD was dependent on intact maternal B cell immunity. Nursing alone, when done by wild-type mothers with AAD, was sufficient for offspring to acquire the antigen-specific protective factor(s) from breastmilk. PMID- 19301987 TI - Sustained effect of the aromatase inhibitors anastrozole and letrozole on endometrial thickness in patients with endometrial hyperplasia and endometrial carcinoma. AB - OBJECTIVE: To investigate whether there may be a role for aromatase inhibitors (AIs) in the treatment of endometrial hyperplasia (EH) and endometrial adenocarcinoma (EA) in postmenopausal women, a retrospective study on the effect of aromatase inhibitors (anastrozole or letrozole) was conducted for 16 patients who were not amenable to surgical treatment. MAIN OUTCOME MEASURE: Resolution of endometrial thickening measured by transvaginal ultrasound at 3-month intervals; the response of metastases was assessed by standard oncological criteria. RESULTS: In all, 16 patients were studied. The BMI of 13 of the 16 patients was known and ranged from 20.7 to 47.7 (mean 34.5) kg/m(2). During treatment with AIs, mean endometrial thickness in the eight patients with EH decreased progressively by 81.7% from 14.7 mm at the start of treatment to 2.7 mm following 36 months of treatment. A greater original mean endometrial thickness of 17 mm was seen in the four patients with localised EA, this fell progressively by 67.1% to 5.6 mm following 36 months of treatment. No responses were seen in four patients with metastatic disease. CONCLUSION: Our results indicate that treatment of EH with anastrozole or letrozole can reduce endometrial thickness as seen ultrasonically, and that in some cases AI treatment can reduce endometrial thickness in patients with localised EA. We found no evidence to indicate that AI treatment prevents disease progression in patients with metastatic EA. Further investigations will be necessary to validate our findings from this small retrospective study and to compare AI inhibitor treatment with topical progestogen therapy. PMID- 19301988 TI - Change in formulation and its potential clinical and pharmacoeconomic value: example of extended release venlafaxine. AB - INTRODUCTION: The 505(b)(2) route of a New Drug Application (NDA) allows published literature or previous FDA findings of safety and effectiveness to be used for approval. Such drugs are not therapeutic equivalents (i.e., generics); instead, the FDA calls them pharmaceutical alternatives. A recent example is the approval of venlafaxine extended-release (ER) tablets, developed as an alternative to the widely used ER venlafaxine capsules. The smaller size of the tablets makes them available in a 225-mg strength, which is the approved maximum dose in major depressive disorder after up-titration but currently unavailable in the capsule formulation, requiring patients on this dose to take two or three capsules; in addition, the tablets are priced at a discount compared to the capsules. METHODS: The objective of this review was to investigate how the change in formulation of ER venlafaxine from capsules to tablets, as an example of such a change in formulation, can potentially offer value to patients and society, with a specific focus on pill burden, drug cost, and adherence. Based on a MEDLINE literature search, the pertinent literature was reviewed in a qualitative manner. REVIEW OF THE LITERATURE: Simplifying treatment regimens, reducing pill burden, and reducing drug costs are recognized strategies for improving adherence. This can be of particular benefit in psychiatric illness because of high rates of nonadherence to treatment. Lack of adherence may negatively impact treatment outcomes and increase disease cost. As such, the ER venlafaxine tablets have the potential to reduce pill burden, improve adherence and outcomes, and reduce cost to patients and society. These preliminary findings need to be corroborated with more primary research and a systematic review of formulation changes. CONCLUSION: A change in formulation of established therapies such as ER venlafaxine has the potential to offer clinical and pharmacoeconomic benefits to patients and society. PMID- 19301989 TI - Tolerability of tapentadol immediate release in patients with lower back pain or osteoarthritis of the hip or knee over 90 days: a randomized, double-blind study. AB - OBJECTIVE: Tapentadol is a novel, centrally acting analgesic with two mechanisms of action, mu-opioid receptor agonism and norepinephrine reuptake inhibition, in a single molecule. This phase III, randomized, double-blind, active-controlled study evaluated the tolerability of tapentadol immediate release (IR) and oxycodone IR for low back pain or osteoarthritis pain (hip or knee), using flexible dosing over 90 days. METHODS: Patients (N = 878) were randomly assigned (4:1 ratio) to receive tapentadol IR (50 or 100 mg, q4-6h, p.o.) or oxycodone IR (10 or 15 mg, q4-6h, p.o.). Tapentadol IR was evaluated for tolerability over 90 days, tolerability relative to oxycodone IR, withdrawal symptoms, and pain intensity. This study was not placebo-controlled, which limited efficacy evaluations. RESULTS: In total, 849 intent-to-treat patients received tapentadol IR (n = 679) or oxycodone IR (n = 170), and among these, 391 patients (57.6%) in the tapentadol IR group and 86 patients (50.6%) in the oxycodone IR group completed the study. Gastrointestinal events, including nausea (18.4% vs 29.4%), vomiting (16.9% vs 30.0%), and constipation (12.8% vs 27.1%), were reported by 44.2% of patients receiving tapentadol IR and 63.5% of patients receiving oxycodone IR, respectively. Nervous system events, including dizziness (18.1% vs 17.1%), headache (11.5% vs 10.0%), and somnolence (10.2% vs 9.4%), were reported by 36.7% of patients receiving tapentadol and 37.1% of patients receiving oxycodone, respectively. Odds ratios (tapentadol:oxycodone) showed that the incidences of somnolence and dizziness were similar; however, nausea, vomiting, and constipation were significantly less likely with tapentadol IR compared with oxycodone IR. The pattern of withdrawal symptoms suggests that drug tapering may not be necessary after tapentadol IR treatment of this duration. Pain intensity measurements showed similar efficacy for tapentadol and oxycodone. CONCLUSION: During this 90-day study, tapentadol IR was associated with improved gastrointestinal tolerability compared with oxycodone IR while providing similar pain relief. Trial registration information: NCT00364546. PMID- 19301990 TI - The first does the work, but the third time's the charm: the effects of massed repetition on episodic encoding of multimodal face-name associations. AB - In social interactions, it is often necessary to rapidly encode the association between visually presented faces and auditorily presented names. The present study used event-related potentials to examine the neural correlates of associative encoding for multimodal face-name pairs. We assessed study-phase processes leading to high-confidence recognition of correct pairs (and consistent rejection of recombined foils) as compared to lower-confidence recognition of correct pairs (with inconsistent rejection of recombined foils) and recognition failures (misses). Both high- and low-confidence retrieval of face-name pairs were associated with study-phase activity suggestive of item-specific processing of the face (posterior inferior temporal negativity) and name (fronto-central negativity). However, only those pairs later retrieved with high confidence recruited a sustained centro-parietal positivity that an ancillary localizer task suggested may index an association-unique process. Additionally, we examined how these processes were influenced by massed repetition, a mnemonic strategy commonly employed in everyday situations to improve face-name memory. Differences in subsequent memory effects across repetitions suggested that associative encoding was strongest at the initial presentation, and thus, that the initial presentation has the greatest impact on memory formation. Yet, exploratory analyses suggested that the third presentation may have benefited later memory by providing an opportunity for extended processing of the name. Thus, although encoding of the initial presentation was critical for establishing a strong association, the extent to which processing was sustained across subsequent immediate (massed) presentations may provide additional encoding support that serves to differentiate face-name pairs from similar (recombined) pairs by providing additional encoding opportunities for the less dominant stimulus dimension (i.e., name). PMID- 19301991 TI - Neural systems underlying lexical competition: an eye tracking and fMRI study. AB - The present study investigated the neural bases of phonological onset competition using an eye tracking paradigm coupled with fMRI. Eighteen subjects were presented with an auditory target (e.g., beaker) and a visual display containing a pictorial representation of the target (e.g., beaker), an onset competitor (e.g., beetle), and two phonologically and semantically unrelated objects (e.g., shoe, hammer). Behavioral results replicated earlier research showing increased looks to the onset competitor compared to the unrelated items. fMRI results showed that lexical competition induced by shared phonological onsets recruits both frontal structures and posterior structures. Specifically, comparison between competitor and no-competitor trials elicited activation in two nonoverlapping clusters in the left IFG, one located primarily within BA 44 and the other primarily located within BA 45, and one cluster in the left supramarginal gyrus (SMG) extending into the posterior superior temporal gyrus. These results indicate that the left IFG is sensitive to competition driven by phonological similarity and not only to competition among semantic/conceptual factors. Moreover, they indicate that the SMG is not only recruited in tasks requiring access to lexical form but is also recruited in tasks that require access to the conceptual representation of a word. PMID- 19301992 TI - Auditory spatial and object processing in the human planum temporale: no evidence for selectivity. AB - Although it is generally acknowledged that at least two processing streams exist in the primate cortical auditory system, the function of the posterior dorsal stream is a topic of much debate. Recent studies have reported selective activation to auditory spatial change in portions of the human planum temporale (PT) relative to nonspatial stimuli such as pitch changes or complex acoustic patterns. However, previous work has suggested that the PT may be sensitive to another kind of nonspatial variable, namely, the number of auditory objects simultaneously presented in the acoustic signal. The goal of the present fMRI experiment was to assess whether any portion of the PT showed spatial selectivity relative to manipulations of the number of auditory objects presented. Spatially sensitive regions in the PT were defined by comparing activity associated with listening to an auditory object (speech from a single talker) that changed location with one that remained stationary. Activity within these regions was then examined during a nonspatial manipulation: increasing the number of objects (talkers) from one to three. The nonspatial manipulation modulated activity within the "spatial" PT regions. No region within the PT was found to be selective for spatial or object processing. We suggest that previously documented spatial sensitivity in the PT reflects auditory source separation using spatial cues rather than spatial processing per se. PMID- 19301993 TI - Visual and semantic processing of living things and artifacts: an FMRI study. AB - We carried out an fMRI study with a twofold purpose: to investigate the relationship between networks dedicated to semantic and visual processing and to address the issue of whether semantic memory is subserved by a unique network or by different subsystems, according to semantic category or feature type. To achieve our goals, we administered a word-picture matching task, with within category foils, to 15 healthy subjects during scanning. Semantic distance between the target and the foil and semantic domain of the target-foil pairs were varied orthogonally. Our results suggest that an amodal, undifferentiated network for the semantic processing of living things and artifacts is located in the anterolateral aspects of the temporal lobes; in fact, activity in this substrate was driven by semantic distance, not by semantic category. By contrast, activity in ventral occipito-temporal cortex was driven by category, not by semantic distance. We interpret the latter finding as the effect exerted by systematic differences between living things and artifacts at the level of their structural representations and possibly of their lower-level visual features. Finally, we attempt to reconcile contrasting data in the neuropsychological and functional imaging literature on semantic substrate and category specificity. PMID- 19301994 TI - Going their separate ways: dissociation of hippocampal and dorsolateral prefrontal activation during episodic retrieval and post-retrieval processing. AB - Hippocampal activity is modulated during episodic memory retrieval. Most consistently, a relative increase in activity during confident retrieval is observed. Dorsolateral prefrontal cortex (DLPFC) is also activated during retrieval, but may be more generally activated during cognitive-control processes. The "default network," regions activated during rest or internally focused tasks, includes the hippocampus, but not DLPFC. Therefore, DLPFC and the hippocampus should diverge during difficult tasks suppressing the default network. It is unclear, however, whether a difficult episodic memory retrieval task would suppress the default network due to difficulty or activate it due to internally directed attention. We hypothesized that a task requiring episodic retrieval followed by rumination on the retrieved item would increase DLPFC activity, but paradoxically reduce hippocampal activity due to concomitant suppression of the default network. In the present study, blocked and event related fMRI were used to examine hippocampal activity during episodic memory recollection and postretrieval processing of paired associates. Subjects were asked to make living/nonliving judgments about items visually presented (classify) or items retrieved from memory (recall-classify). Active and passive baselines were used to differentiate task-related activity from default-network activity. During the "recall-classify" task, anterior hippocampal activity was selectively reduced relative to "classify" and baseline tasks, and this activity was inversely correlated with DLPFC. Reaction time was positively correlated with DLPFC activation and default-network/hippocampal suppression. The findings demonstrate that frontal and hippocampal activity are dissociated during difficult episodic retrieval tasks and reveal important considerations for interpreting hippocampal activity associated with successful episodic retrieval. PMID- 19301995 TI - Effective connectivity of the fronto-parietal network during attentional control. AB - The ACC, the dorsolateral prefrontal cortex (DLPFC), and the parietal cortex near/along the intraparietal sulcus (IPS) are members of a network subserving attentional control. Our recent study revealed that these regions participate in both response anticipation and conflict processing. However, little is known about the relative contribution of these regions in attentional control and how the dynamic interactions among these regions are modulated by detection of predicted versus unpredicted targets and conflict processing. Here, we examined effective connectivity using dynamic causal modeling among these three regions during a flanker task with or without a target onset cue. We compared various models in which different connections among ACC, DLPFC, and IPS were modulated by bottom-up stimulus-driven surprise and top-down conflict processing using Bayesian model selection procedures. The most optimal of these models incorporated contextual modulation that allowed processing of unexpected (surprising) targets to mediate the influence of the IPS over ACC and DLPFC and conflict processing to mediate the influence of ACC and DLPFC over the IPS. This result suggests that the IPS plays an initiative role in this network in the processing of surprise targets, whereas ACC and DLPFC interact with each other to resolve conflict through attentional modulation implemented via the IPS. PMID- 19301996 TI - On the processing of semantic aspects of experience in the anterior medial temporal lobe: an event-related fMRI study. AB - Recognition memory based on familiarity judgments is a form of declarative memory that has been repeatedly associated with the anterior medial temporal lobe. It has been argued that this region sustains familiarity-based recognition not only by retrieving item-specific information but also by coding for those semantic aspects of an event that support later familiarity-based recognition. Here, we used event-related fMRI to directly examine whether the contribution of anterior medial temporal lobe to declarative memory indeed results from its role in processing semantic aspects of an event. For this purpose, a sentence comprehension task was employed which varied the demands of semantic and syntactic processing of the sentence-final word. By presenting those sentence final words together with new words in a subsequent incidental recognition memory test, we were able to determine the mnemonic consequences of presenting words in different sentential contexts. Results showed that enhanced semantic processing during comprehension activates regions in medial temporal lobe cortex and leads to response suppression in partly overlapping regions when the word is successfully retrieved. Data from a behavioral follow-up study support the view that enhanced semantic processing at study enhances familiarity-based remembering in a subsequent test phase. PMID- 19301997 TI - Memory repression: brain mechanisms underlying dissociative amnesia. AB - Dissociative amnesia usually follows a stressful event and cannot be attributable to explicit brain damage. It is thought to reflect a reversible deficit in memory retrieval probably due to memory repression. However, the neural mechanisms underlying this condition are not clear. We used fMRI to investigate neural activity associated with memory retrieval in two patients with dissociative amnesia. For each patient, three categories of face photographs and three categories of people's names corresponding to the photographs were prepared: those of "recognizable" high school friends who were acquainted with and recognizable to the patients, those of "unrecognizable" colleagues who were actually acquainted with but unrecognizable to the patients due to their memory impairments, and "control" distracters who were unacquainted with the patients. During fMRI, the patients were visually presented with these stimuli and asked to indicate whether they were personally acquainted with them. In the comparison of the unrecognizable condition with the recognizable condition, we found increased activity in the pFC and decreased activity in the hippocampus in both patients. After treatment for retrograde amnesia, the altered pattern of brain activation disappeared in one patient whose retrograde memories were recovered, whereas it remained unchanged in the other patient whose retrograde memories were not recovered. Our findings provide direct evidence that memory repression in dissociative amnesia is associated with an altered pattern of neural activity, and they suggest the possibility that the pFC has an important role in inhibiting the activity of the hippocampus in memory repression. PMID- 19301998 TI - Visual short-term memory capacity for simple and complex objects. AB - Does the capacity of visual short-term memory (VSTM) depend on the complexity of the objects represented in memory? Although some previous findings indicated lower capacity for more complex stimuli, other results suggest that complexity effects arise during retrieval (due to errors in the comparison process with what is in memory) that is not related to storage limitations of VSTM, per se. We used ERPs to track neuronal activity specifically related to retention in VSTM by measuring the sustained posterior contralateral negativity during a change detection task (which required detecting if an item was changed between a memory and a test array). The sustained posterior contralateral negativity, during the retention interval, was larger for complex objects than for simple objects, suggesting that neurons mediating VSTM needed to work harder to maintain more complex objects. This, in turn, is consistent with the view that VSTM capacity depends on complexity. PMID- 19301999 TI - The contribution of primary motor cortex is essential for probabilistic implicit sequence learning: evidence from theta burst magnetic stimulation. AB - Theta burst transcranial magnetic stimulation (TBS) is considered to produce plastic changes in human motor cortex. Here, we examined the inhibitory and excitatory effects of TBS on implicit sequence learning using a probabilistic serial reaction time paradigm. We investigated the involvement of several cortical regions associated with implicit sequence learning by examining probabilistic sequence learning in five age- and IQ-matched groups of healthy participants following continuous inhibitory TBS over primary motor cortex (M1); or the supplementary motor area (SMA) or dorsolateral prefrontal cortex (DLPFC) or following intermittent excitatory TBS of M1; or after sham TBS. Relative to sham TBS, probabilistic sequence learning was abolished by inhibitory TBS over M1, demonstrating that this region is critical for implicit motor sequence learning. Sequence learning was not significantly affected by inhibitory TBS over the SMA, DLPFC or excitatory TBS over M1. These results demonstrate that the M1 mediates implicit sequence learning. PMID- 19302000 TI - Target detection by opponent coding in monkey prefrontal cortex. AB - The pFC plays a key role in flexible, context-specific decision making. One proposal [Machens, C. K., Romo, R., & Brody, C. D. Flexible control of mutual inhibition: A neural model of two-interval discrimination. Science, 307, 1121 1124, 2005] is that prefrontal cells may be dynamically organized into opponent coding circuits, with competitive groups of cells coding opposite behavioral decisions. Here, we show evidence for extensive, temporally evolving opponent organization in the monkey pFC during a cued target detection task. More than a half of all randomly selected cells discriminated stimulus category in this task. The largest set showed target-positive activity, with the strongest responses to the current target, intermediate activity for a nontarget that was a target on other trials, and lowest activity for nontargets never associated with the target category. Second most frequent was a reverse, antitarget pattern. In the ventrolateral frontal cortex, opponent organization was strongly established in phasic responses at stimulus onset; later, such activity was widely spread across dorsolateral and ventrolateral sites. Task-specific organization into opponent cell groups may be a general feature of prefrontal decision making. PMID- 19302002 TI - Rapid detection of emotion from human vocalizations. AB - The rapid detection of affective signals from conspecifics is crucial for the survival of humans and other animals; if those around you are scared, there is reason for you to be alert and to prepare for impending danger. Previous research has shown that the human brain detects emotional faces within 150 msec of exposure, indicating a rapid differentiation of visual social signals based on emotional content. Here we use event-related brain potential (ERP) measures to show for the first time that this mechanism extends to the auditory domain, using human nonverbal vocalizations, such as screams. An early fronto-central positivity to fearful vocalizations compared with spectrally rotated and thus acoustically matched versions of the same sounds started 150 msec after stimulus onset. This effect was also observed for other vocalized emotions (achievement and disgust), but not for affectively neutral vocalizations, and was linked to the perceived arousal of an emotion category. That the timing, polarity, and scalp distribution of this new ERP correlate are similar to ERP markers of emotional face processing suggests that common supramodal brain mechanisms may be involved in the rapid detection of affectively relevant visual and auditory signals. PMID- 19302001 TI - Investigating occipito-temporal contributions to reading with TMS. AB - The debate regarding the role of ventral occipito-temporal cortex (vOTC) in visual word recognition arises, in part, from difficulty delineating the functional contributions of vOTC as separate from other areas of the reading network. Here, we investigated the feasibility of using TMS to interfere with vOTC processing in order to explore its specific contributions to visual word recognition. Three visual lexical decision experiments were conducted using neuronavigated TMS. The first demonstrated that repetitive stimulation of vOTC successfully slowed word, but not nonword, responses. The second confirmed and extended these findings by demonstrating the effect was specific to vOTC and not present in the adjacent lateral occipital complex. The final experiment used paired-pulse TMS to investigate the time course of vOTC processing for words and revealed activation starting as early as 80-120 msec poststimulus onset significantly earlier than that expected based on electrophysiological and magnetoencephalography studies. Taken together, these results clearly indicate that TMS can be successfully used to stimulate parts of vOTC previously believed to be inaccessible and provide a new tool for systematically investigating the information processing characteristics of vOTC. In addition, the findings provide strong evidence that lexical status and frequency significantly affect vOTC processing, findings difficult to reconcile with prelexical accounts of vOTC function. PMID- 19302003 TI - Lesions to the motor system affect action perception. AB - Several studies have shown that the motor system is involved in action perception, suggesting that action concepts are represented through sensory-motor processes. Such conclusions imply that motor system impairments should diminish action perception. To test this hypothesis, a group of 10 brain-damaged patients with hemiplegia (specifically, a lesion at the motor system that affected the contralesional arm) viewed point-light displays of arm gestures and attempted to name each gesture. To create the dynamic stimuli, patients individually performed simple gestures with their unaffected arm while being videotaped. The videotapes were converted into point-light animations. Each action was presented as it had been performed, that is, as having been produced by the observer's unaffected arm, and in its mirror reversed orientation, that is, as having been produced by the observer's hemiplegic arm. Action recognition accuracy by patients with hemiplegia was compared with that by 8 brain-damaged patients without any motor deficit and by 10 healthy controls. Overall, performance was better in control observers than in patients. Most importantly, performance by hemiplegic patients, but not by nonhemiplegic patients and controls, varied systematically as a function of the observed limb. Action recognition was best when hemiplegic patients viewed actions that appeared to have been performed by their unaffected arm. Action recognition performance dropped significantly when hemiplegic patients viewed actions that appeared to have been produced with their hemiplegic arm or the corresponding arm of another person. The results of a control study involving the recognition of point-light defined animals in motion indicate that a generic deficit to visual and cognitive functions cannot account for this laterality-specific deficit in action recognition. Taken together, these results suggest that motor cortex impairment decreases visual sensitivity to human action. Specifically, when a cortical lesion renders an observer incapable of performing an observed action, action perception is compromised, possibly by a failure to map the observed action onto the observer's contralesional hemisoma. PMID- 19302004 TI - Emergent oscillations in evolutionary simulations: oscillating networks increase switching efficacy. AB - LeDoux [LeDoux, J. E. The emotional brain. New York: Simon & Schuster, 1996] motivated the direct route in his dual-pathway model by arguing that the ability to switch rapidly between different modes of behavior is highly adaptive. This motivation was supported by evolutionary simulations [den Dulk, P., Heerebout, B. T., & Phaf, R. H. A computational study into the evolution of dual-route dynamics for affective processing. Journal of Cognitive Neuroscience, 15, 194-208, 2003], in which foraging agents, controlled by simple inheritable neural networks, navigated an artificial world while avoiding predation. After many generations, a dual-processing architecture evolved that enabled a rapid switch to avoidance behavior when a predator appeared. We added recurrent connections to a new "context" layer in the indirect pathway to provide the agents with a working memory of previous input (i.e., a "context"). Unexpectedly, agents with oscillating networks emerged that had a much higher fitness than agents without oscillations. Oscillations seemed to have similar effects on switching speed as the dual-processing architecture, but they enhanced switching efficacy to a much larger degree. There has been much neurobiological speculation on the function, if any, of neural oscillations. These findings suggest that the facilitation of switching behavior is a likely candidate. Moreover, the strongly improved adaptation in the simulations contradicts the position that neural oscillations are merely a by-product of cell firing and have no functional value [Pareti, G., & De Palma, A. Does the brain oscillate? The dispute on neuronal synchronization. Neurological Sciences, 25, 41-47, 2004]. PMID- 19302005 TI - Auditory working memory load impairs visual ventral stream processing: toward a unified model of attentional load. AB - Attentional interference between tasks performed in parallel is known to have strong and often undesired effects. As yet, however, the mechanisms by which interference operates remain elusive. A better knowledge of these processes may facilitate our understanding of the effects of attention on human performance and the debilitating consequences that disruptions to attention can have. According to the load theory of cognitive control, processing of task-irrelevant stimuli is increased by attending in parallel to a relevant task with high cognitive demands. This is due to the relevant task engaging cognitive control resources that are, hence, unavailable to inhibit the processing of task-irrelevant stimuli. However, it has also been demonstrated that a variety of types of load (perceptual and emotional) can result in a reduction of the processing of task irrelevant stimuli, suggesting a uniform effect of increased load irrespective of the type of load. In the present study, we concurrently presented a relevant auditory matching task [n-back working memory (WM)] of low or high cognitive load (1-back or 2-back WM) and task-irrelevant images at one of three object visibility levels (0%, 50%, or 100%). fMRI activation during the processing of the task-irrelevant visual stimuli was measured in the lateral occipital cortex and found to be reduced under high, compared to low, WM load. In combination with previous findings, this result is suggestive of a more generalized load theory, whereby cognitive load, as well as other types of load (e.g., perceptual), can result in a reduction of the processing of task-irrelevant stimuli, in line with a uniform effect of increased load irrespective of the type of load. PMID- 19302006 TI - Perception of face parts and face configurations: an FMRI study. AB - fMRI studies have reported three regions in human ventral visual cortex that respond selectively to faces: the occipital face area (OFA), the fusiform face area (FFA), and a face-selective region in the superior temporal sulcus (fSTS). Here, we asked whether these areas respond to two first-order aspects of the face argued to be important for face perception, face parts (eyes, nose, and mouth), and the T-shaped spatial configuration of these parts. Specifically, we measured the magnitude of response in these areas to stimuli that (i) either contained real face parts, or did not, and (ii) either had veridical face configurations, or did not. The OFA and the fSTS were sensitive only to the presence of real face parts, not to the correct configuration of those parts, whereas the FFA was sensitive to both face parts and face configuration. Further, only in the FFA was the response to configuration and part information correlated across voxels, suggesting that the FFA contains a unified representation that includes both kinds of information. In combination with prior results from fMRI, TMS, MEG, and patient studies, our data illuminate the functional division of labor in the OFA, FFA, and fSTS. PMID- 19302007 TI - Audiovisual matching in speech and nonspeech sounds: a neurodynamical model. AB - Audiovisual speech perception provides an opportunity to investigate the mechanisms underlying multimodal processing. By using nonspeech stimuli, it is possible to investigate the degree to which audiovisual processing is specific to the speech domain. It has been shown in a match-to-sample design that matching across modalities is more difficult in the nonspeech domain as compared to the speech domain. We constructed a biophysically realistic neural network model simulating this experimental evidence. We propose that a stronger connection between modalities in speech underlies the behavioral difference between the speech and the nonspeech domain. This could be the result of more extensive experience with speech stimuli. Because the match-to-sample paradigm does not allow us to draw conclusions concerning the integration of auditory and visual information, we also simulated two further conditions based on the same paradigm, which tested the integration of auditory and visual information within a single stimulus. New experimental data for these two conditions support the simulation results and suggest that audiovisual integration of discordant stimuli is stronger in speech than in nonspeech stimuli. According to the simulations, the connection strength between auditory and visual information, on the one hand, determines how well auditory information can be assigned to visual information, and on the other hand, it influences the magnitude of multimodal integration. PMID- 19302008 TI - Meningococcal factor H-binding protein variants expressed by epidemic capsular group A, W-135, and X strains from Africa. AB - BACKGROUND: Meningococcal epidemics in Africa are generally caused by capsular group A strains, but W-135 or X strains also cause epidemics in this region. Factor H-binding protein (fHbp) is a novel antigen being investigated for use in group B vaccines. Little is known about fHbp in strains from other capsular groups. METHODS: We investigated fHbp in 35 group A, W-135, and X strains from Africa. RESULTS: The 22 group A isolates, which included each of the sequence types (STs) responsible for epidemics since 1963, and 4 group X and 3 group W-135 isolates from recent epidemics had genes encoding fHbp in antigenic variant group 1. The remaining 6 W-135 isolates had fHbp variant 2. Within each fHbp variant group, there was 92%-100% amino acid identity, and the proteins expressed conserved epitopes recognized by bactericidal monoclonal antibodies. Serum samples obtained from mice vaccinated with native outer membrane vesicle vaccines from mutants engineered to express fHbp variants had broad bactericidal activity against group A, W-135, or X strains. CONCLUSIONS: Despite extensive natural exposure of the African population, fHbp is conserved among African strains. A native outer membrane vesicle vaccine that expresses fHbp variants can potentially elicit protective antibodies against strains from all capsular groups that cause epidemics in the region. PMID- 19302009 TI - Free-living amoebae as agents of human infection. PMID- 19302010 TI - Molecular confirmation of Sappinia pedata as a causative agent of amoebic encephalitis. AB - Pathogenic free-living amoebae, such as Acanthamoeba species, Balamuthia mandrillaris, and Naegleria fowleri, are known to cause infections of the central nervous system in human and other animals. In 2001, a case of human encephalitis was reported that was caused by another amoeba with morphological features suggestive of Sappinia. The amoeba originally identified as Sappinia diploidea was identified, most likely as S. pedata, by use of newly developed real-time polymerase chain reaction assays. This amoeba had previously been found only in environmental sources, such as soil and tree bark. The results illustrate the potential for other free-living amoebae, which are not normally associated with human disease, to cause occasional infections. PMID- 19302011 TI - Combination of host susceptibility and virulence of Mycobacterium tuberculosis determines dual role of nitric oxide in the protection and control of inflammation. AB - Tuberculosis (TB) remains a global health threat. Although it is generally accepted that TB results from intensive cross-talk between the host and the pathogen Mycobacterium tuberculosis, underlying mechanisms remain elusive. The first evidence of human polymorphisms related to susceptibilities to distinct M. tuberculosis lineages has been gathered. Confrontation of limited host resistance with heightened bacterial virulence forms a most hazardous combination. We investigated extreme combinations, confronting inducible nitric oxide synthase deficient (iNOS(-/-)) and wild-type (WT) mice with 2 related M. tuberculosis strains that differ markedly in virulence, namely, the M. tuberculosis laboratory strains H37Rv and H37Ra. We provide evidence that deregulated chemokine signaling and excessive neutrophil necrosis contribute to disproportionate neutrophil influx and exacerbated TB in iNOS(-/-) mice infected with virulent M. tuberculosis (strain H37Rv), whereas resistant and susceptible mice controlled attenuated H37Ra equally well. Thus, a combination of host susceptibility and M. tuberculosis virulence determines the role of iNOS in the protection and control of inflammation. PMID- 19302012 TI - Cytokine profiles in toxoplasmic and viral uveitis. AB - BACKGROUND: Uveitis is a major cause of visual impairment throughout the world. Analysis of cytokine profiles in aqueous humor specimens may provide insight into the physiopathological processes that underly retinal damage in this context. METHODS: Using a multiplex assay, we determined the concentrations of 17 cytokines and chemokines in aqueous humor specimens obtained from patients with ocular toxoplasmosis or viral uveitis and compared these concentrations with those in specimens obtained from patients with noninfectious intermediate uveitis or cataract. RESULTS: Five mediators (interleukin [IL]-8, monocyte chemoattractant protein-1, tumor necrosis factor-alpha, IL-4, and IL-10) were detected in >50% of patients in all groups. In contrast, IL-5 and IL-12 were specific for ocular toxoplasmosis, and granulocyte monocyte colony-stimulating factor and IL-1 were specific for viral uveitis; these mediators could present specific markers for diagnostic purposes. Interferon-gamma, IL-6, and macrophage inflammatory protein-1beta were common markers of ocular toxoplasmosis and viral uveitis. IL-17 was a common marker of ocular toxoplasmosis and intermediate uveitis. CONCLUSIONS: We found specific cytokine profiles for each type of uveitis, with large interindividual variations and no etiological or clinical correlations. Ocular cytokine mapping contributes to a better understanding of the physiopathology of specific forms of uveitis and provides guidance for new targeted treatment. PMID- 19302014 TI - DAS181 and H5N1 virus infection. PMID- 19302015 TI - Comparison between single-diode low-level laser therapy (LLLT) and LED multi diode (cluster) therapy (LEDT) applications before high-intensity exercise. AB - BACKGROUND DATA AND OBJECTIVE: There is anecdotal evidence that low-level laser therapy (LLLT) may affect the development of muscular fatigue, minor muscle damage, and recovery after heavy exercises. Although manufacturers claim that cluster probes (LEDT) maybe more effective than single-diode lasers in clinical settings, there is a lack of head-to-head comparisons in controlled trials. This study was designed to compare the effect of single-diode LLLT and cluster LEDT before heavy exercise. MATERIALS AND METHODS: This was a randomized, placebo controlled, double-blind cross-over study. Young male volleyball players (n = 8) were enrolled and asked to perform three Wingate cycle tests after 4 x 30 sec LLLT or LEDT pretreatment of the rectus femoris muscle with either (1) an active LEDT cluster-probe (660/850 nm, 10/30 mW), (2) a placebo cluster-probe with no output, and (3) a single-diode 810-nm 200-mW laser. RESULTS: The active LEDT group had significantly decreased post-exercise creatine kinase (CK) levels ( 18.88 +/- 41.48 U/L), compared to the placebo cluster group (26.88 +/- 15.18 U/L) (p < 0.05) and the active single-diode laser group (43.38 +/- 32.90 U/L) (p < 0.01). None of the pre-exercise LLLT or LEDT protocols enhanced performance on the Wingate tests or reduced post-exercise blood lactate levels. However, a non significant tendency toward lower post-exercise blood lactate levels in the treated groups should be explored further. CONCLUSION: In this experimental set up, only the active LEDT probe decreased post-exercise CK levels after the Wingate cycle test. Neither performance nor blood lactate levels were significantly affected by this protocol of pre-exercise LEDT or LLLT. PMID- 19302016 TI - Clinical significance and outcome of nosocomial acquisition of spontaneous bacterial peritonitis in patients with liver cirrhosis. AB - BACKGROUND: There have been few reports on the causes and treatment outcomes for nosocomial spontaneous bacterial peritonitis (SBP) in patients with liver cirrhosis. METHODS: We performed a retrospective cohort study to compare the microbiological and clinical characteristics in nosocomial versus community acquired SBP. All patients with SBP, for whom culture was proven to be positive for SBP at Samsung Medical Center (Seoul, Republic of Korea) from 1 January 2000 through 31 June 2007, were included. Medical records and laboratory data were reviewed. Nosocomial SBP was defined as SBP diagnosed after 72 h of hospitalization. RESULTS: A total of 236 patients with SBP were enrolled (mean age +/- SD age, 56.6 +/- 10.7 years); 166 patients were women, and 70 were men. Nosocomial and community-acquired SBP occurred in 126 and 110 patients, respectively. Escherichia coli accounted for 102 (43.2%) of 236 isolates, Klebsiella species accounted for 33 isolates (14.0%), and Streptococcus species accounted for 23 isolates (9.8%). The overall 30-day mortality rate for nosocomial SBP was higher than that for community-acquired SBP (58.7% vs. 37.3%; P = .001). Nosocomial isolates of gram-negative organisms were significantly more resistant to third-generation cephalosporins (41% vs. 10.0%; P = .001) and quinolones (50.0% vs. 30.9%; P = .003), compared with community-acquired isolates. Multivariate analysis revealed that nosocomial infection, concomitant hepatocellular carcinoma, presentation with acute renal failure or shock, and resistance to third-generation cephalosporins were significant risk factors for 30-day mortality associated with SBP. CONCLUSIONS: Nosocomial SBP has a poorer outcome than community-acquired SBP. The resistance to third-generation cephalosporins for gram-negative organisms, which are more common in nosocomial cases of SBP than in community-acquired cases of SBP, adversely affects the outcome of SBP in patients with liver cirrhosis. PMID- 19302017 TI - Changes in body composition with ritonavir-boosted and unboosted atazanavir treatment in combination with Lamivudine and Stavudine: a 96-week randomized, controlled study. AB - This 96-week, open-label, randomized study assessed changes in body composition in treatment-naive patients infected with human immunodeficiency virus type 1 who were treated with either atazanavir or ritonavir-boosted atazanavir, in combination with stavudine and lamivudine. Both treatment groups had similar increases in trunk fat, but patients treated with ritonavir-boosted atazanavir had a significantly lower incidence of lipoatrophy. PMID- 19302018 TI - Genital and perianal herpes simplex simulating neoplasia in patients with AIDS. AB - Genital and perianal herpetic ulcers are common in HIV-infected patients and chronic mucocutaneous ulcers persisting for more than 1 month are the hallmark of active AIDS status. However, atypical clinical manifestations of herpes simplex virus (HSV) may occur in immunocompromised patients presenting as tumor-like nodules or condylomatous or hypertrophic lesions, rather than a classic ulcer. Such unusual presentations raise the risk of misdiagnosis and a delay in appropriate treatment. Here we describe nine immunocompromised HIV-positive patients with CD 4 count ranging from 14-362/mm(3) (mean 170/mm(3)), with unusual tumoral presentation of anogenital herpes. There were six male and three female patients with AIDS with mean duration of HIV infection of 14 years. All of the patients had history of highly active antiretroviral therapy (HAART), with five patients compliant with the therapy at the time of biopsy. Six patients presented with scrotal or vulvar masses and three with perianal nodules. Five patients had adjacent human papilloma virus (HPV)-related lesions. Prior to excision, herpetic lesion was clinically suspected in only three patients and in the rest of the patients a malignant growth was the main clinical concern. The predominant histopathologic finding was dense dermal plasmacytic infiltration with overlying pseudoepitheliomatous hyperplasia, superficial ulcers and classic herpetic inclusions. Patients with AIDS may experience excessive number and size of both primary and reactivated herpetic lesions. The tumoral presentations discussed here are less common, but are often clinically misdiagnosed. It is important to be aware of these unusual presentations to provide a correct diagnosis and prompt, effective treatment for HSV. Several studies suggest that aggressive treatment of HSV in combination with HAART therapy provides a significant survival benefit. Pathobiology mechanisms of unusual and exaggerated tumor-like inflammatory response are not completely elucidated. PMID- 19302019 TI - Generic combination approved. PMID- 19302020 TI - More than $1.17 million funds hepatitis C awareness, prevention, and care. PMID- 19302021 TI - Lymphedema, at the forefront. PMID- 19302022 TI - Recent advances in breast cancer-related lymphedema of the arm: lymphatic pump failure and predisposing factors. AB - Axillary surgery for breast cancer may be followed, months to years later, by chronic arm lymphedema. A simple 'stopcock' mechanism (reduced lymph drainage from the entire limb through surviving lymphatics) does not explain many clinical aspects, including the delayed onset and selective sparing of some regions, e.g., hand. Quantitative lymphoscintigraphy reveals that lymph drainage is slowed in the subcutis, where most of the edema lies, and in the subfascial muscle compartment, which normally has much higher lymph flows than the subcutis. Although the muscle does not swell significantly, the impaired muscle drainage correlates with the severity of arm swelling, indicating a likely key role for muscle lymphatic function. A new method, lymphatic congestion lymphoscintigraphy, showed that the edema is associated with a reduced contractility of the arm lymphatics; the weaker the active lymphatic pump, the greater the swelling. Delayed lymphatic pump failure may result from chronic raised afterload, as in hypertensive cardiac failure, and may account for the delayed onset of swelling. A further novel finding is that lymph flow is raised in both the subcutis and muscle of both arms in postsurgical breast patients who later developed breast cancer-related lymphedema (BCRL), compared with patients who did not develop BCRL. This new observation indicates a predisposition to BCRL in some women. Further evidence for predisposing abnormalities is the finding of lymphatic abnormalities in the contralateral (nonswollen) arm in women with established BCRL. Such predisposing factors could explain why some women develop BCRL after sentinel node biopsy, whereas others do not after clearance surgery. Future research must focus on prospective observations made from before surgery until BCRL develops. PMID- 19302023 TI - Therapeutic responses to exogenous VEGF-C administration in experimental lymphedema: immunohistochemical and molecular characterization. AB - BACKGROUND: In a widely employed murine tail model of human acquired lymphedema, we have previously observed that, distal to the site of experimental lymphatic ablation, there is immunohistochemical evidence of a profound increase in cutaneous lymphatic vessel number and size that normalizes after VEGF-C administration. OBJECTIVE: In order to investigate the mechanistic basis of the lymphatic microvascular remodeling, we have studied the lymphedematous responses to VEGF-C after co-administration of systemic VEGFR-3 neutralizing antibody. We have also undertaken genome-wide whole-tissue transcriptional profiling of lymphedematous tissues before and after exogenous VEGF-C administration. STUDY DESIGN: We provoked postsurgical lymphedema in the mouse tail model and assessed the effects of exogenously administered human recombinant VEGF-C in the presence of a monoclonal anti-VEGFR-3 antibody. Polyclonal IgG was administered to a series of control subjects. Microvascular lymphatic remodeling was assessed through quantitative and qualitative anti-LYVE1 immunohistochemistry. Genome-wide transcriptional profiling was performed in whole skin derived from lymphedema with and without exogenous VEGF-C administration. Normal mice and surgical shams served as controls. RESULTS: In the presence of the monoclonal anti-VEGFR-3 neutralizing antibody, positive lymphatic microvascular remodeling in lymphedematous skin is nearly completely abrogated. Furthermore, the therapeutic impact of added VEGF-C is markedly attenuated, as is the ability of the growth factor to ameliorate tissue edema. Transcriptional profiling of the VEGF-C responses in treated lymphedema reveals a very restricted list of genes whose expression is upregulated in lymphedema and re-normalized following VEGF-C treatment. CONCLUSION: The postsurgical murine tail model of lymphedema closely simulates attributes of human lymphedema. The current series of investigations underscores the utility of the murine tail model to the preclinical and translational investigation of lymphedema. The derived insights continue to focus favorably upon the central role of the VEGFR-3 receptor and its ligands in the development and therapeutic resolution of lymphedema. Whole tissue transcriptional profiling continues to shed light on disease mechanisms and potential future targets for therapeutic intervention. PMID- 19302026 TI - Energy availability and density estimates in African ungulates. AB - While many studies have investigated the link between primary productivity and species richness, the link between primary productivity and species abundance is still poorly understood. We explored how primary productivity, assessed by the satellite-based normalized difference vegetation index, influenced density estimates in 13 African ungulate species. We found that primary productivity generally correlated positively with density estimates. However, this link was more pronounced for some species than for others, and information regarding the area surveyed needed to be taken into account to highlight this functional relationship. Detailed consideration of measurements of productivity in broadscale studies identify a general link between primary productivity and abundance of African ungulates and highlight remote-sensing-based analyses as a potentially crucial tool in assessing the population consequences of future changes in primary productivity on this continent. PMID- 19302027 TI - Ecological limits on clade diversification in higher taxa. AB - Species richness varies dramatically among groups of organisms, yet the causes of this variation remain poorly understood. Variation in species-level diversification rates may partially explain differential species richness among clades, but older clades should also be more diverse, because they will have had more time to accumulate species. Surprisingly, studies that have investigated this question have reached dramatically different conclusions: several claim to find no such age-diversity relationship, whereas a recent and more inclusive study reported that clade age and not diversification rate explains the variation in species richness among animal taxa. Here I address the relationship between clade age and species richness using a model-based approach that controls for variation in diversification rates among clades. I find that species richness is effectively independent of clade age in four of five data sets. Even extreme among-clade variation in diversification rates cannot account for the absence of a positive age-diversity relationship in angiosperms, birds, and teleost fishes. I consider two alternative explanations for these results and find that a clade volatility model positing correlated speciation-extinction dynamics does not underlie these patterns. Rather, ecological limits on clade growth, such as geographic area, appear to mediate temporal declines in diversification within higher taxa. PMID- 19302028 TI - Patterns of phenotypic plasticity in common and rare environments: a study of host use and color learning in the cabbage white butterfly Pieris rapae. AB - Phenotypic plasticity is adaptive in variable environments but, given its costs, may be disfavored if only one environment is commonly encountered. Yet species in relatively constant environments often adjust phenotypes successfully in rare or novel environments. Developmental biases may reduce the costs of plasticity in common environments, favoring the maintenance of plasticity. We explored this proposition by studying the flexibility of visually guided host-selection behavior in cabbage white butterflies (Pieris rapae), wherein common and rare environments consisted of green and red host types, respectively. We demonstrated in greenhouse assays that adult females display an innate bias toward green color during host search but alter that bias through learning in red-host assemblages such that, after several hours of experience, red hosts are located as efficiently as green hosts. Full-sib analyses suggested there was genetic variation in host and color choice that was more pronounced in the red-host environment. We found no evidence of genetic correlations in behavior across host environments or of fitness costs of plasticity in color choice. Our results support the idea that learning may persist in less variable environments through the evolution of innate biases that reduce operating costs in common environments. PMID- 19302029 TI - No trade-off between seed size and number in the valley oak Quercus lobata. AB - We examined the relationship between acorn mass and number in valley oaks (Quercus lobata) over 4 years in central coastal California. Despite considerable variation in acorn size among both trees and years, trees produced acorns of the same size relative to other trees in different years. Across years, the relationship between acorn mass and acorn crop size was generally positive, even after controlling for environmental conditions and differences in individual tree size and quality. Life-history trade-offs in valley oaks are primarily between current and future reproduction and indirectly between concurrent growth and reproduction, not between seed size and number, and are probably related to this species' mast-seeding behavior. Phenotypic trade-offs in long-lived plants such as oaks exhibit complex patterns of life-history covariation and deserve greater attention, both theoretically and empirically. PMID- 19302030 TI - Variation in local density results in a positive correlation between plant neighbor sizes. AB - A positive correlation between the sizes of plants and the sizes of their nearest neighbors has been observed in many plant populations and is most often attributed to variation in microsite quality. We demonstrate another potentially important cause, using a simple spatial simulation model in which a plant's size is determined by its available area plus stochastic variation. If the spatial arrangement of individuals is nonuniform and local crowding reduces individual size, a positive correlation between the size of a plant and the size of neighboring individuals is to be expected. If a plant is small because it is crowded, its nearest neighbors will also tend to be crowded and therefore small. Such positive correlations will be observed whenever any trait of an object is related to the area available to it. Looking at the performance of neighboring objects without explicitly considering distance can give a very misleading signal. PMID- 19302031 TI - Harmful mating tactics in hermaphrodites. AB - While empirical data suggest that sperm competition and multiple mating both contribute to the evolution of harmful mating tactics in hermaphrodites, a precise understanding of their interaction is lacking. We therefore formulate a game-theoretical model of mating behavior in hermaphrodites, where harmful mating tactics confer an advantage in sperm competition while simultaneously reducing the mating partner's survival. The model predicts evolutionarily stable values of resource allocation between sexual functions and the degree of harmful mating. Our analysis provides support for the empirical observation that harmful mating is associated with multiply mating species in which sperm precedence strongly favors the first mate. The model also shows that this criterion becomes less important as harmful mating tactics become more efficient. As harmful tactics make sperm displacement more effective, a consequence is a more female-biased resource allocation. Provided that fertilized egg production is not limited by availability of sperm, a more female-biased allocation should increase the number of offspring produced, but the model instead shows that harmful mating tactics more than countercompensate, leading to reduced fitness. Hermaphrodites that use harmful mating tactics may therefore be at a disadvantage when competing with other species for a limited resource. PMID- 19302032 TI - Metabolic alkalosis due to feeding chicks in breeding Adelie penguins Pygoscelis adeliae under natural conditions. AB - Prolonged abnormal vomiting causes metabolic alkalosis. Many seabirds are known to feed their chicks by regurgitation. We hypothesized that metabolic alkalosis occurs in seabirds even under natural conditions during the breeding season. Adelie penguins Pygoscelis adeliae feed their chicks by regurgitating food for 50 60 d until the chicks fledge. In this study, the concentrations of Cl(-), HCO(3)( ), Na+, K+, pH, and PCO2 in the blood of breeding Adelie penguins were measured throughout the chick-rearing season. The pH of penguin venous blood shifted from 7.54 in the guarding period to 7.47 in the creche period. Decreasing Cl(-) and increasing HCO(3)(-) blood concentrations in parents were associated with increasing mass of their brood in the guarding period, the early phase of the rearing season, indicating that regurgitating to feed chicks causes loss of gastric acid and results in relative metabolic alkalosis. The inverse trend was observed during the creche period, the latter phase of the rearing season, when parents spent more time at sea and have fewer opportunities for gastric acid loss. This was assumed to be the recovery phase. These results indicate that regurgitation might cause metabolic alkalosis in breeding Adelie penguins. To our knowledge, this is the first report to indicate that seabirds exhibit metabolic alkalosis due to regurgitation to feed chicks under natural conditions. PMID- 19302033 TI - Locally applied osteogenic predifferentiated progenitor cells are more effective than undifferentiated mesenchymal stem cells in the treatment of delayed bone healing. AB - Mesenchymal stem cells (MSCs) and osteogenic predifferentiated cells (OPCs) have been shown to promote healing of critical-sized bone defects. This study investigated the regenerative capacity of autologous MSCs versus OPCs after local injection into a compromised bone healing situation. We hypothesized that treatment with MSCs and OPCs would enhance the healing and that the MSCs would be more effective due to their lower differentiation and higher proliferative competence. The femur of rats was osteotomized and stabilized with an external fixator. Except for the control group (C group), in all animals a delayed healing was induced by cautering the periosteum and removing the bone marrow. Two days postsurgery, autologous MSCs (MSC group), OPCs (OPC group), or cell-free medium (Sham group) was percutaneously injected into the osteotomy gap. The C group received no treatment. Bone healing was evaluated radiologically, biomechanically, and histologically. After 8 weeks, the C group showed complete bony bridging, while a delayed healing was detected in the Sham group. All outcome measures showed better healing of the OPC group compared to the Sham group. Contrary to our expectations, there were no significant differences in outcome measures between the MSC group and the Sham group. The percutaneous injection of OPCs could become a minimally invasive treatment option for delayed or nonunions. PMID- 19302034 TI - MYC quantitation in cell-free plasma DNA by real-time PCR for gastric cancer diagnosis. AB - BACKGROUND: Detection of tumor-associated genetic alterations in plasma of cancer patients has recently been suggested to be an accurate method for detecting early or recurrent cancer. METHODS: We performed quantitative real-time PCR for MYC and GAPDH in tissue and plasma samples of 57 patients with gastric cancer and in plasma of 79 cancer-free individuals. We also performed two-color MYC fluorescence in situ hybridization (FISH) in tissue from the 57 patients with gastric cancer. RESULTS: The tissue MYC/GAPDH ratio by real-time PCR was significantly correlated with MYC status by FISH (p<0.001). The mean ratio of plasma MYC/GAPDH was 5.226+/-3.578 (range: 1.25-18.35) in gastric cancer patients, and 2.436+/-0.881 (range: 1.00-5.00) in the healthy volunteers (p<0.001). We used receiver-operating characteristics (ROC) curve analysis to select two optimal plasma MYC/GAPDH cut-offs of 2.725 and 5.225. The sensitivity and specificity were 75.4% and 76.9% at 2.725, 38.6% and 100% at 5.225, respectively. The plasma MYC/GAPDH ratio from cancer patients was significantly correlated with the tissue MYC/GAPDH ratio (p=0.009), and tissue MYC status by FISH (p=0.024). CONCLUSIONS: These findings suggest that the plasma MYC/GAPDH ratio, as determined by real-time PCR, may be an alternative non-invasive approach for detecting gastric cancer. PMID- 19302035 TI - A routine method for the quantification of the novel antimycotic drug posaconazole in plasma using liquid chromatography-tandem mass spectrometry. AB - BACKGROUND: Posaconazole is now widely used for prophylaxis of invasive fungal infections in immunocompromised patients. The pharmacokinetic properties of the drug argue for therapeutic monitoring, but so far described analytical methods have shortcomings with respect to application in a routine setting. The aim of our work was to develop an analytical method suitable for routine use. METHODS: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used. For sample preparation, protein precipitation followed by on-line solid phase extraction was used. SCH 56984, a posaconazole related compound provided by the manufacturer of posaconazole, was used as internal standard. RESULTS: The method was characterized by short hands-on time and an analytical run time of four minutes. Validation data demonstrated acceptable analytical performance. CONCLUSIONS: LC MS/MS with on-line solid phase extraction for sample preparation allows the implementation of a convenient and reliable method for pharmacokinetic monitoring of posaconazole. PMID- 19302036 TI - Microglial physiology: unique stimuli, specialized responses. AB - Microglia, the macrophages of the central nervous system parenchyma, have in the normal healthy brain a distinct phenotype induced by molecules expressed on or secreted by adjacent neurons and astrocytes, and this phenotype is maintained in part by virtue of the blood-brain barrier's exclusion of serum components. Microglia are continually active, their processes palpating and surveying their local microenvironment. The microglia rapidly change their phenotype in response to any disturbance of nervous system homeostasis and are commonly referred to as activated on the basis of the changes in their morphology or expression of cell surface antigens. A wealth of data now demonstrate that the microglia have very diverse effector functions, in line with macrophage populations in other organs. The term activated microglia needs to be qualified to reflect the distinct and very different states of activation-associated effector functions in different disease states. Manipulating the effector functions of microglia has the potential to modify the outcome of diverse neurological diseases. PMID- 19302037 TI - The liver as a lymphoid organ. AB - The liver receives blood from both the systemic circulation and the intestine, and in distinctive, thin-walled sinusoids this mixture passes over a large macrophage population, termed Kupffer cells. The exposure of liver cells to antigens, and to microbial products derived from the intestinal bacteria, has resulted in a distinctive local immune environment. Innate lymphocytes, including both natural killer cells and natural killer T cells, are unusually abundant in the liver. Multiple populations of nonhematopoietic liver cells, including sinusoidal endothelial cells, stellate cells located in the subendothelial space, and liver parenchymal cells, take on the roles of antigen-presenting cells. These cells present antigen in the context of immunosuppressive cytokines and inhibitory cell surface ligands, and immune responses to liver antigens often result in tolerance. Important human pathogens, including hepatitis C virus and the malaria parasite, exploit the liver's environment, subvert immunity, and establish persistent infection. PMID- 19302038 TI - Immune and inflammatory mechanisms of atherosclerosis (*). AB - Atherosclerosis is an inflammatory disease of the wall of large- and medium-sized arteries that is precipitated by elevated levels of low-density lipoprotein (LDL) cholesterol in the blood. Although dendritic cells (DCs) and lymphocytes are found in the adventitia of normal arteries, their number is greatly expanded and their distribution changed in human and mouse atherosclerotic arteries. Macrophages, DCs, foam cells, lymphocytes, and other inflammatory cells are found in the intimal atherosclerotic lesions. Beneath these lesions, adventitial leukocytes organize in clusters that resemble tertiary lymphoid tissues. Experimental interventions can reduce the number of available blood monocytes, from which macrophages and most DCs and foam cells are derived, and reduce atherosclerotic lesion burden without altering blood lipids. Under proatherogenic conditions, nitric oxide production from endothelial cells is reduced and the burden of reactive oxygen species (ROS) and advanced glycation end products (AGE) is increased. Incapacitating ROS-generating NADPH oxidase or the receptor for AGE (RAGE) has beneficial effects. Targeting inflammatory adhesion molecules also reduces atherosclerosis. Conversely, removing or blocking IL-10 or TGF-beta accelerates atherosclerosis. Regulatory T cells and B1 cells secreting natural antibodies are atheroprotective. This review summarizes our current understanding of inflammatory and immune mechanisms in atherosclerosis. PMID- 19302039 TI - Primary B cell immunodeficiencies: comparisons and contrasts. AB - Sophisticated genetic tools have made possible the identification of the genes responsible for most well-described immunodeficiencies in the past 15 years. Mutations in Btk, components of the pre-B cell and B cell receptor (lambda5, Igalpha, Igbeta), or the scaffold protein BLNK account for approximately 90% of patients with defects in early B cell development. Hyper-IgM syndromes result from mutations in CD40 ligand, CD40, AID, or UNG in 70-80% of affected patients. Rare defects in ICOS or CD19 can result in a clinical picture that is consistent with common variable immunodeficiency, and as many as 10% of patients with this disorder have heterozygous amino acid substitutions in TACI. For all these disorders, there is considerable clinical heterogeneity in patients with the same mutation. Identifying the genetic and environmental factors that influence the clinical phenotype may enhance patient care and our understanding of normal B cell development. PMID- 19302040 TI - The inflammasomes: guardians of the body. AB - The innate immune system relies on its capacity to rapidly detect invading pathogenic microbes as foreign and to eliminate them. The discovery of Toll-like receptors (TLRs) provided a class of membrane receptors that sense extracellular microbes and trigger antipathogen signaling cascades. More recently, intracellular microbial sensors have been identified, including NOD-like receptors (NLRs). Some of the NLRs also sense nonmicrobial danger signals and form large cytoplasmic complexes called inflammasomes that link the sensing of microbial products and metabolic stress to the proteolytic activation of the proinflammatory cytokines IL-1beta and IL-18. The NALP3 inflammasome has been associated with several autoinflammatory conditions including gout. Likewise, the NALP3 inflammasome is a crucial element in the adjuvant effect of aluminum and can direct a humoral adaptive immune response. In this review, we discuss the role of NLRs, and in particular the inflammasomes, in the recognition of microbial and danger components and the role they play in health and disease. PMID- 19302041 TI - Human marginal zone B cells. AB - Human marginal zone (MZ) B cells are, in a sense, a new entity. Although they share many properties with their mouse counterpart, they also display striking differences, such as the capacity to recirculate and the presence of somatic mutations in their B cell receptor. These differences are the reason they are often not considered a separate, rodent-like B cell lineage, but rather are considered IgM memory B cells. We review here our present knowledge concerning this subset and the arguments in favor of the proposition that humans have evolved for their MZ B cell compartment a separate B cell population that develops and diversifies its Ig receptor during ontogeny outside T-dependent or T independent immune responses. PMID- 19302042 TI - Aire. AB - Mutations in the transcriptional regulator, Aire, cause APECED, a polyglandular autoimmune disease with monogenic transmission. Animal models of APECED have revealed that Aire plays an important role in T cell tolerance induction in the thymus, mainly by promoting ectopic expression of a large repertoire of transcripts encoding proteins normally restricted to differentiated organs residing in the periphery. The absence of Aire results in impaired clonal deletion of self-reactive thymocytes, which escape into the periphery and attack a variety of organs. In addition, Aire is a proapoptotic factor, expressed at the final maturation stage of thymic medullary epithelial cells, a function that may promote cross-presentation of the antigens encoded by Aire-induced transcripts in these cells. Transcriptional regulation by Aire is unusual in being very broad, context-dependent, probabilistic, and noisy. Structure/function analyses and identification of its interaction partners suggest that Aire may impact transcription at several levels, including nucleosome displacement during elongation and transcript splicing or other aspects of maturation. PMID- 19302043 TI - Regulatory lymphocytes and intestinal inflammation. AB - The immune system is pivotal in mediating the interactions between host and microbiota that shape the intestinal environment. Intestinal homeostasis arises from a highly dynamic balance between host protective immunity and regulatory mechanisms. This regulation is achieved by a number of cell populations acting through a set of shared regulatory pathways. In this review, we summarize the main lymphocyte subsets controlling immune responsiveness in the gut and their mechanisms of control, which involve maintenance of intestinal barrier function and suppression of chronic inflammation. CD4(+)Foxp3(+) T cells play a nonredundant role in the maintenance of intestinal homeostasis through IL-10- and TGF-beta-dependent mechanisms. Their activity is complemented by other T and B lymphocytes. Because breakdown in immune regulatory networks in the intestine leads to chronic inflammatory diseases of the gut, such as inflammatory bowel disease and celiac disease, regulatory lymphocytes are an attractive target for therapies of intestinal inflammation. PMID- 19302044 TI - The ins and outs of leukocyte integrin signaling. AB - Integrins are the principal cell adhesion receptors that mediate leukocyte migration and activation in the immune system. These receptors signal bidirectionally through the plasma membrane in pathways referred to as inside-out and outside-in signaling. Each of these pathways is mediated by conformational changes in the integrin structure. Such changes allow high-affinity binding of the receptor with counter-adhesion molecules on the vascular endothelium or extracellular matrix and lead to association of the cytoplasmic tails of the integrins with intracellular signaling molecules. Leukocyte functional responses resulting from outside-in signaling include migration, proliferation, cytokine secretion, and degranulation. Here, we review the key signaling events that occur in the inside-out versus outside-in pathways, highlighting recent advances in our understanding of how integrins are activated by a variety of stimuli and how they mediate a diverse array of cellular responses. PMID- 19302045 TI - Recent advances in the genetics of autoimmune disease. AB - Extraordinary technical advances in the field of human genetics over the past few years have catalyzed an explosion of new information about the genetics of human autoimmunity. In particular, the ability to scan the entire genome for common polymorphisms that associate with disease has led to the identification of numerous new risk genes involved in autoimmune phenotypes. Several themes are emerging. Autoimmune disorders have a complex genetic basis; multiple genes contribute to disease risk, each with generally modest effects independently. In addition, it is now clear that common genes underlie multiple autoimmune disorders. There is also heterogeneity among subphenotypes within a disease and across major racial groups. The current crop of genetic associations are only the start of a complete catalog of genetic factors for autoimmunity, and it remains unclear to what extent common variation versus multiple rare variants contribute to disease susceptibility. The current review focuses on recent discoveries within functionally related groups of genes that provide clues to novel pathways of pathogenesis for human autoimmunity. PMID- 19302047 TI - Immunological and inflammatory functions of the interleukin-1 family. AB - More than any other cytokine family, the interleukin (IL)-1 family is closely linked to the innate immune response. This linkage became evident upon the discovery that the cytoplasmic domain of the IL-1 receptor type I is highly homologous to the cytoplasmic domains of all Toll-like receptors (TLRs). Thus, fundamental inflammatory responses such as the induction of cyclooxygenase type 2, increased expression of adhesion molecules, or synthesis of nitric oxide are indistinguishable responses of both IL-1 and TLR ligands. Both families nonspecifically affect antigen recognition and lymphocyte function. IL-1beta is the most studied member of the IL-1 family because of its role in mediating autoinflammatory diseases. Although the TLR and IL-1 families evolved to assist in host defense against infection, unlike the TLR family, the IL-1 family also includes members that suppress inflammation, both specifically within the IL-1 family but also nonspecifically for TLR ligands and the innate immune response. PMID- 19302046 TI - Cell-mediated immune responses in tuberculosis. AB - Tuberculosis is primarily a disease of the lung, and dissemination of the disease depends on productive infection of this critical organ. Upon aerosol infection with Mycobacterium tuberculosis (Mtb), the acquired cellular immune response is slow to be induced and to be expressed within the lung. This slowness allows infection to become well established; thus, the acquired response is expressed in an inflammatory site that has been initiated and modulated by the bacterium. Mtb has a variety of surface molecules that interact with the innate response, and this interaction along with the autoregulation of the immune response by several mechanisms results in less-than-optimal control of bacterial growth. To improve current vaccine strategies, we must understand the factors that mediate induction, expression, and regulation of the immune response in the lung. We must also determine how to induce both known and novel immunoprotective responses without inducing immunopathologic consequences. PMID- 19302048 TI - Regulatory T cells in the control of host-microorganism interactions (*). AB - Each microenvironment requires a specific set of regulatory elements that are finely and constantly tuned to maintain local homeostasis. Various populations of regulatory T cells contribute to the maintenance of this equilibrium and establishment of controlled immune responses. In particular, regulatory T cells limit the magnitude of effector responses, which may result in failure to adequately control infection. However, regulatory T cells also help limit collateral tissue damage caused by vigorous antimicrobial immune responses against pathogenic microbes as well as commensals. In this review, we describe various situations in which the balance between regulatory T cells and effector immune functions influence the outcome of host-microorganism coexistence and discuss current hypotheses and points of polemic associated with the origin, target, and antigen specificity of both endogenous and induced regulatory T cells during these interactions. PMID- 19302050 TI - Regulation and function of NF-kappaB transcription factors in the immune system. AB - The mammalian Rel/NF-kappaB family of transcription factors, including RelA, c Rel, RelB, NF-kappaB1 (p50 and its precursor p105), and NF-kappaB2 (p52 and its precursor p100), plays a central role in the immune system by regulating several processes ranging from the development and survival of lymphocytes and lymphoid organs to the control of immune responses and malignant transformation. The five members of the NF-kappaB family are normally kept inactive in the cytoplasm by interaction with inhibitors called IkappaBs or the unprocessed forms of NF kappaB1 and NF-kappaB2. A wide variety of signals emanating from antigen receptors, pattern-recognition receptors, receptors for the members of TNF and IL 1 cytokine families, and others induce differential activation of NF-kappaB heterodimers. Although work over the past two decades has shed significant light on the regulation of NF-kappaB transcription factors and their functions, much progress has been made in the past two years revealing new insights into the regulation and functions of NF-kappaB. This recent progress is covered in this review. PMID- 19302052 TI - Atraumatic osteonecrosis of the humeral head. AB - While much literature has focused on the management of osteonecrosis of the femoral head, far less information is available regarding the treatment and outcomes of this disease in the proximal humerus. To a great extent, management of humeral head osteonecrosis has been inferred from studies involving the femoral head. The etiologies for this disease can be categorized most usefully as traumatic versus atraumatic. Regardless of the underlying etiology, the common pathway involves disruption of the arterial inflow or the venous outflow of the bone, with resultant osseous cell death. The general treatment strategies for humeral head osteonecrosis include nonoperative modalities for symptomatic early disease, with surgical intervention reserved for more advanced disease or those with recalcitrant pain. PMID- 19302053 TI - Tendon transfers for irreparable rotator cuff tears. PMID- 19302054 TI - Patellofemoral instability. AB - This review describes the normal patellofemoral joint and detail the mechanism and anatomic elements that predispose patients to patellar instability. The treatment options for both acute and chronic injuries are described and the rationale behind their approach to this problem is explained. In general, most acute dislocations should be treated nonoperatively unless the instability is associated with an osteochondral injury. Chronic dislocators should be treated based on an understanding of the patient's individual reason for recurrent instability. This is achieved with a thorough history, physical examination, and imaging studies. This information can help the clinician select the most appropriate proximal and or distal procedure. PMID- 19302049 TI - Horror autoinflammaticus: the molecular pathophysiology of autoinflammatory disease (*). AB - The autoinflammatory diseases are characterized by seemingly unprovoked episodes of inflammation, without high-titer autoantibodies or antigen-specific T cells. The concept was proposed ten years ago with the identification of the genes underlying hereditary periodic fever syndromes. This nosology has taken root because of the dramatic advances in our knowledge of the genetic basis of both mendelian and complex autoinflammatory diseases, and with the recognition that these illnesses derive from genetic variants of the innate immune system. Herein we propose an updated classification scheme based on the molecular insights garnered over the past decade, supplanting a clinical classification that has served well but is opaque to the genetic, immunologic, and therapeutic interrelationships now before us. We define six categories of autoinflammatory disease: IL-1beta activation disorders (inflammasomopathies), NF-kappaB activation syndromes, protein misfolding disorders, complement regulatory diseases, disturbances in cytokine signaling, and macrophage activation syndromes. A system based on molecular pathophysiology will bring greater clarity to our discourse while catalyzing new hypotheses both at the bench and at the bedside. PMID- 19302055 TI - Patellofemoral arthritis. AB - Patellofemoral joint arthritis is a common condition which can be extremely debilitating. Although it is a common condition, the treatment of isolated patellofemoral arthritis varies and remains controversial. The objective of this review is to provide an overview of the current understanding of patellofemoral arthritis and various different surgical options, indications, and their respective outcomes. PMID- 19302056 TI - Acute syndesmosis injuries associated with ankle fractures: current perspectives in management. AB - Ankle syndesmosis injuries frequently occur with ankle fractures, but their treatment remains controversial. Although specific clinical and radiographic diagnostic measures are generally well-accepted, there remains a lack of consensus with respect to the treatment of these injuries. Controversy arises at almost every phase of treatment including: type of fixation (screw size, type of implant), number of cortices required for fixation, and need for hardware removal. Regardless of fixation technique chosen, the most important goal should be anatomic reduction and restoration of the syndesmosis and ankle mortise as this is the only significant predictor of functional outcome. PMID- 19302057 TI - Posterior cruciate ligament injuries in the athlete: diagnosis and treatment. AB - Posterior cruciate ligament injuries occur much less frequently than anterior cruciate ligament injuries. We review the important physical examination and radiographic findings, as well as provide the indications for nonoperative and operative treatment. PMID- 19302058 TI - Midshaft clavicle fractures in adults. AB - Clavicle fractures remain one of the more common fractures encountered in the orthopaedic office. Nonoperative management remains the standard of care in most cases. However, studies have shown a higher nonunion rate and decreased patient satisfaction in fractures with increased shortening. The malunion resulting from nonoperative management has been shown to be problematic in patients with significant fracture shortening. When operative reduction and fixation is indicated, there are numerous techniques to aid the surgeon patient care. This article reviews the midshaft clavicle fracture and discusses recent outcome studies on patients with fracture shortening and approaches to operative management. PMID- 19302059 TI - Timing issues in fracture management: a review of current concepts. AB - The timing of operative fracture care is often confounded by multisystem injuries, conflicting or absent literature, and lack of communication between orthopaedic surgeons and other physicians providing care to the patient. Much has been published regarding the proper sequence of events in providing care to patients with multisystem injuries. Only recently has the role of complex musculoskeletal injuries and the timing of fixation in multiply-injured patients been explored in detail. Timing of care for pelvic injuries is frequently determined by the presence of injury to other organ systems, the presence of open wounds, and hemodynamic status. There is likely an optimal time window for fixation. However, existing data is often difficult to compare, given varying definitions and protocols. Furthermore, reports are often conflicting, making the determination of an optimal time-window difficult. Similar concerns are present with lower extremity long bone fractures. Injury to other organ systems must be considered with timing of femur fixation, particularly in the presence of lung injury. Tibia fractures are frequently complicated by the presence of a tenuous soft tissue envelope and other injury factors that often alter the timing of fixation. These issues and, last, the timing of care for calcaneus and talus injuries are reviewed, as risk of avascular necrosis and quality of articular reduction are related to the timing of fixation. PMID- 19302060 TI - Evaluation and treatment of injuries of the ulnar collateral ligament of the thumb metacarpophalangeal joint. AB - Injuries of the ulnar collateral ligament of the metacarpophalangeal joint of the thumb are very common. A review of the literature regarding the pertinent anatomy, injury mechanism, diagnostic modalities, and treatment options may yield a better understanding for future management. PMID- 19302061 TI - Surface replacement arthroplasty of the hip. AB - Treatment of the young patient with degenerative disease of the hip has historically been a difficult problem for the orthopaedist. Total hip arthroplasty in the young patient has generally produced inferior results as compared to older patients. Surface replacement arthroplasty (SRA) was initially developed over 50 years ago to treat degenerative disease of the hip. It has regained enthusiasm over the last 10 to 15 years as an alternative to total hip arthroplasty for the treatment of degenerative disease of the hip in younger patients. The modern metal-on-metal bearing provides improved wear characteristics over its metal-on-polyethylene predecessor. Multiple studies have demonstrated mid-term results of metal-on-metal SRA, which are comparable to total hip arthroplasty. The long-term survival data of SRA remains to be seen, as does the long-term effect of elevated serum ion levels. PMID- 19302062 TI - Brachial plexus birth palsy: an overview of early treatment considerations. AB - Since the description by Smellie in 1764, in a French midwifery text, that first suggested an obstetric origin for upper limb birth palsy, great strides have been made in both diagnosis and early and late treatment. This report presents an overview of selected aspects of this complex and extensive subject. Early treatment options are reviewed in the context of the present controversies regarding the natural history and the indications for and timing of microsurgical intervention in infants with brachial plexus birth injuries. PMID- 19302063 TI - The distal radioulnar joint. AB - The distal radioulnar joint (DRUJ) acts in concert with the proximal radioulnar joint to control forearm rotation. The DRUJ is stabilized by the triangular fibrocartilage complex (TFCC). This complex of fibrocartilage and ligaments support the joint through its arc of rotation, as well as provide a smooth surface for the ulnar side of the carpus. TFCC and DRUJ injuries are part of the common pattern of injuries we see with distal radius fractures. While much attention has been paid to the treatment of the distal radius fractures, many of the poor outcomes are due to untreated or unrecognized injuries to the DRUJ and its components. PMID- 19302064 TI - Modulation of the hepatic malonyl-CoA-carnitine palmitoyltransferase 1A partnership creates a metabolic switch allowing oxidation of de novo fatty acids. AB - Liver mitochondrial beta-oxidation of LCFAs (long-chain fatty acids) is tightly regulated through inhibition of CPT1A (carnitine palmitoyltransferase 1A) by malonyl-CoA, an intermediate of lipogenesis stimulated by glucose and insulin. Moreover, CPT1A sensitivity to malonyl-CoA inhibition varies markedly depending on the physiopathological state of the animal. In the present study, we asked whether an increase in CPT1A activity solely or in association with a decreased malonyl-CoA sensitivity could, even in the presence of high glucose and insulin concentrations, maintain a sustained LCFA beta-oxidation and/or protect from triacylglycerol (triglyceride) accumulation in hepatocytes. We have shown that adenovirus-mediated expression of rat CPT1wt (wild-type CPT1A) and malonyl-CoA insensitive CPT1mt (CPT1AM593S mutant) in cultured fed rat hepatocytes counteracted the inhibition of oleate beta-oxidation induced by 20 mM glucose/10 nM insulin. Interestingly, the glucose/insulin-induced cellular triacylglycerol accumulation was prevented, both in the presence and absence of exogenous oleate. This resulted from the generation of a metabolic switch allowing beta-oxidation of de novo synthesized LCFAs, which occurred without alteration in glucose oxidation and glycogen synthesis. Moreover, CPT1mt expression was more effective than CPT1wt overexpression to counteract glucose/insulin effects, demonstrating that control of CPT1A activity by malonyl-CoA is an essential driving force for hepatic LCFA metabolic fate. In conclusion, the present study highlights that CPT1A is a prime target to increase hepatic LCFA beta-oxidation and that acting directly on the degree of its malonyl-CoA sensitivity may be a relevant strategy to prevent and/or correct hepatic steatosis. PMID- 19302065 TI - Guidelines for the management of contact dermatitis: an update. AB - These guidelines for management of contact dermatitis have been prepared for dermatologists on behalf of the British Association of Dermatologists. They present evidence-based guidance for investigation and treatment, with identification of the strength of evidence available at the time of preparation of the guidelines, including details of relevant epidemiological aspects, diagnosis and investigation. PMID- 19302066 TI - Dermoscopic and histopathological findings of polymorphous vessels in amelanotic cutaneous metastasis of pigmented cutaneous melanoma. PMID- 19302067 TI - Dermoscopy in the diagnosis of tungiasis. PMID- 19302068 TI - Species of Borrelia burgdorferi complex that cause borrelial lymphocytoma in France. AB - BACKGROUND: Only about 30 cases of borrelial lymphocytoma (BL) with identification of the causative species of Borrelia have been published to date, mainly from Eastern or Central European countries. OBJECTIVES: To identify the species of B. burgdorferi complex responsible for BL in France. METHODS: Nine patients with BL acquired in France and for whom skin samples were sent to the national reference centre laboratory between 1994 and 2007 were included in this retrospective study. Direct detection of Borrelia in skin samples was made by polymerase chain reaction targeting the fla gene. Culture was performed when technically possible, and identification of each species was made by hybridization of a fragment of the fla gene with a panel of species-specific oligonucleotides. RESULTS: Borrelia afzelii was identified in three cases, B. garinii and B. burgdorferi sensu stricto in one case each. Culture was positive in only one case (B. garinii). CONCLUSIONS: Borrelia afzelii seems to be the predominant species of Borrelia responsible for BL in France, as already reported in other European countries. PMID- 19302069 TI - Hand eczema and quality of life: a population-based study. AB - BACKGROUND: Hand eczema is a common disease in the population and is of interest from a public health perspective. Health-related quality of life (HRQoL) is increasingly being measured in dermatology. OBJECTIVES: To investigate HRQoL in relation to hand eczema in the general population. METHODS: In the Public Health Survey of Stockholm County Council 2006, a questionnaire was sent to 57 009 randomly selected individuals aged 18-84 years. The response rate among persons of working age (18-64 years) was 58%. The questionnaire included a validated question concerning hand eczema and a generic instrument for measurement of HRQoL, the EQ-5D. RESULTS: The proportion of individuals reporting problems was significantly larger among those with than without hand eczema in all five dimensions of the EQ-5D. Gender differences were found in some age subgroups. The EQ-5D index was lower for individuals with hand eczema than for those without, and on the same level as for psoriasis and asthma. Beta regression showed that the strongest confounding factors were low back pain, depression and hay fever/asthma. CONCLUSIONS: HRQoL was negatively affected in individuals with hand eczema irrespective of age. With the EQ-5D instrument it is also possible to detect certain gender differences. The EQ-5D index for hand eczema was of the same size as for psoriasis and asthma, all common diseases with an impact on public health. It is of importance to acknowledge the influence of hand eczema on daily life, in order to give the patients good care. PMID- 19302070 TI - Sirolimus-associated lymphoedema: eight new cases and a proposed mechanism. AB - Sirolimus, an inhibitor of the mammalian target of rapamycin (mTOR), is increasingly used as an agent for post-transplant immunosuppression and the treatment of solid organ and haematological malignancies and hamartomas. Its advantages include a lack of nephrotoxicity and a lower incidence of nonmelanoma skin cancers; adverse effects include delayed wound healing, increased lymphocoele formation and rarely lymphoedema. We report a series of eight cases of severe, sustained, unilateral and bilateral lymphoedema in patients receiving sirolimus for post-transplant immunosuppression, classify their lymphoscintigraphy findings and propose a mechanism of aetiology based on the interaction of mTOR with key mediators of lymphangiogenesis. PMID- 19302071 TI - An open pilot study of ambulatory photodynamic therapy using a wearable low irradiance organic light-emitting diode light source in the treatment of nonmelanoma skin cancer. AB - BACKGROUND: Photodynamic therapy (PDT) is a popular treatment for nonmelanoma skin cancer with clearance rates of between 70% and 100%. Although reported to have a superior cosmetic outcome, the inconvenience of hospital visits and discomfort during therapy are considered drawbacks. OBJECTIVES: To present an open pilot study of a low-irradiance, potentially disposable, lightweight, organic light-emitting diode (OLED), which is an area-emitting light source (2 cm diameter), suitable for ambulatory PDT. METHODS: Twelve patients with Bowen's disease (eight) and superficial basal cell carcinoma (four) < 2 cm in diameter were recruited into the study following histological confirmation of the diagnosis. Two treatments (45-60 J cm(-2) red light, 550-750 nm, peak 620 nm, irradiance 5 mW cm(-2)) were administered 1 month apart following application of aminolaevulinic acid for 4 h. RESULTS: At the 12-month follow-up, seven of the 12 patients remained clear, with four of the nonresponders demonstrating peripheral margin failure. Patients were scored for pain during and immediately after treatment using the numerical rating scale (NRS; 1-10). All 12 subjects scored pain as < 2 using the NRS (median score 1). In contrast, a similar cohort of 50 consecutive patients from our routine PDT clinic (Aktilite inorganic LED source; 75 J cm(-2), irradiance 80 mW cm(-2)) scored a median of 6 on the NRS. CONCLUSIONS: Pain and inconvenience are practical barriers to the use of conventional PDT. This pilot study suggests that OLED-PDT is less painful than conventional PDT with the added advantage of being lightweight, and therefore has the potential for more convenient 'home PDT'. These results need to be validated in larger studies. PMID- 19302072 TI - Systematic review of dermoscopy and digital dermoscopy/ artificial intelligence for the diagnosis of melanoma. AB - BACKGROUND: Dermoscopy improves diagnostic accuracy of the unaided eye for melanoma, and digital dermoscopy with artificial intelligence or computer diagnosis has also been shown useful for the diagnosis of melanoma. At present there is no clear evidence regarding the diagnostic accuracy of dermoscopy compared with artificial intelligence. OBJECTIVES: To evaluate the diagnostic accuracy of dermoscopy and digital dermoscopy/artificial intelligence for melanoma diagnosis and to compare the diagnostic accuracy of the different dermoscopic algorithms with each other and with digital dermoscopy/artificial intelligence for the detection of melanoma. METHODS: A literature search on dermoscopy and digital dermoscopy/artificial intelligence for melanoma diagnosis was performed using several databases. Titles and abstracts of the retrieved articles were screened using a literature evaluation form. A quality assessment form was developed to assess the quality of the included studies. Heterogeneity among the studies was assessed. Pooled data were analysed using meta-analytical methods and comparisons between different algorithms were performed. RESULTS: Of 765 articles retrieved, 30 studies were eligible for meta-analysis. Pooled sensitivity for artificial intelligence was slightly higher than for dermoscopy (91% vs. 88%; P = 0.076). Pooled specificity for dermoscopy was significantly better than artificial intelligence (86% vs. 79%; P < 0.001). Pooled diagnostic odds ratio was 51.5 for dermoscopy and 57.8 for artificial intelligence, which were not significantly different (P = 0.783). There were no significance differences in diagnostic odds ratio among the different dermoscopic diagnostic algorithms. CONCLUSIONS: Dermoscopy and artificial intelligence performed equally well for diagnosis of melanocytic skin lesions. There was no significant difference in the diagnostic performance of various dermoscopy algorithms. The three-point checklist, the seven-point checklist and Menzies score had better diagnostic odds ratios than the others; however, these results need to be confirmed by a large-scale high-quality population-based study. PMID- 19302073 TI - Left ventricular mechanical dyssynchrony is load independent at rest and during endotoxaemia in a porcine model. AB - AIM: In diseased or injured states, the left ventricle displays higher degrees of mechanical dyssynchrony. We aimed at assessing mechanical dyssynchrony ranges in health related to variation in load as well as during acute endotoxin-induced ventricular injury. METHODS: In 16 juvenile anaesthetized pigs, a five-segment conductance catheter was placed in the left ventricle as well as a balloon-tipped catheter in the inferior vena cava. Mechanical dyssynchrony during systole, including dyssynchrony time in per cent during systole and internal flow fraction during systole, were measured at rest and during controlled pre-load reduction sequences, as well as during 3 h of endotoxin infusion (0.25 microg kg(-)1 h( 1)). RESULTS: Systolic dyssynchrony and internal flow fraction did not change during the course of acute beat-to-beat pre-load alteration. Endotoxin-produced acute pulmonary hypertension by left ventricular dyssynchrony measures was not changed during the early peak of pulmonary hypertension. Endotoxin ventricular injury led to progressive increases in systolic mechanical segmental dyssynchrony (7.9 +/- 1.2-13.0 +/- 1.3%) and ventricular systolic internal flow fraction (7.1 +/- 2.4-16.6 +/- 2.8%), respectively for baseline and then at hour 3. There was no localization of dyssynchrony changes to segment or region in the ventricular long axis during endotoxin infusion. CONCLUSION: These results suggest that systolic mechanical dyssynchrony measures may be load independent in health and during acute global ventricular injury by endotoxin. More study is needed to validate ranges in health and disease for parameters of mechanical dyssynchrony. PMID- 19302074 TI - Clinical trial: cyclophosphamide pulse therapy - a promising therapeutic alternative in refractory Crohn's disease. AB - BACKGROUND: In severe steroid-refractory Crohn's disease (CD), established therapies fail in a relevant proportion of patients. Recent pilot studies indicated the efficacy of cyclophosphamide pulse therapy in these patients. AIM: To provide further and substantial evidence for the rationale to apply cyclophosphamide pulse therapy as therapeutic option in severe courses of CD. METHODS: Fifteen patients with steroid-refractory (n = 13) or steroid-dependent (n = 2) CD received 2-6 (median 3) monthly pulses of 750 mg cyclophosphamide in an open-label fashion. Eleven patients were on concomitant immunosuppression (azathioprine/mercaptopurine n = 9; methotrexate n = 2). RESULTS: Thirteen of 15 patients (87%) had a clinical response (CDAI decrease >100). Ten patients (67%) went into remission (CDAI <150) after 8 weeks. Steroid-free remission was achieved in eight patients (54%). Two patients (13%) failed to respond. Median CDAI decreased from 420 (245-550) to 100 (26-538) at week 8. Remission lasted 16 months (median, range 4-40). In three patients, arthritis, erythema nodosum and episcleritis completely resolved. Cyclophosphamide pulse therapy administration was well tolerated in all subjects. CONCLUSIONS: Cyclophosphamide pulse therapy is safe and highly effective for induction and maintenance of remission in steroid-refractory/-dependent CD. There is a strong need for additional experience to improve the setting of the encouraging cyclophosphamide treatment in CD. PMID- 19302075 TI - Unilateral chemosis, acute onset myopia and choroidal detachment following the use of tamsulosin. PMID- 19302076 TI - Axonal loss occurs early in dominant optic atrophy. AB - PURPOSE: This study set out to investigate retinal nerve fibre layer (RNFL) thickness and best corrected visual acuity (BCVA) in relation to age in healthy subjects and patients with OPA1 autosomal dominant optic atrophy (DOA). METHODS: We carried out a cross-sectional investigation of RNFL thickness and ganglion cell layer density in 30 healthy subjects and 10 patients with OPA1 DOA using optical coherence tomography (OCT). We then performed a regression analysis of RNFL thickness and BCVA versus age. RESULTS: Both healthy subjects and DOA patients demonstrated a gradual reduction in RNFL thickness with age; the relationship was best described statistically by a model that assumed a constant offset between the two groups. Best corrected VA decreased significantly with age in DOA patients, in whom BCVA was correlated with peripapillary RNFL thickness in the inferior and superior peripapillary quadrants and with total macular thickness at eccentricities of 500-3000 microm. The observations were best described by a constant offset of 41.9 microm separating the two groups and an annual decrease in RNFL thickness of 0.48 microm (p < 0.0001). In patients with DOA, increasing age was associated with decreasing BCVA (p = 0.046). CONCLUSIONS: This cross-sectional study found evidence of comparable age-related decreases in RNFL thickness in healthy subjects and in DOA patients, where the deficit in DOA patients is best described using a model that assumes the deficit between the groups does not vary with age. The gradual reduction of BCVA with age may be a consequence of a relative deficit in RNFL thickness that is established before the second decade of life. PMID- 19302077 TI - Viscoanaesthesia in cataract surgery: a prospective, randomized clinical trial. AB - PURPOSE: We aimed to compare viscoanaesthesia (VisThesia) with intracameral lidocaine in cataract surgery carried out under topical anaesthesia. METHODS: In this prospective study 98 patients were randomly assigned to receive VisThesia (group 1, n = 49) or 0.5 cc of 1% unpreserved lidocaine (group 2, n = 49). All surgery was carried out by one surgeon using clear corneal technique. Pachymetry, the status of the cornea and anterior chamber, and intraocular pressure (IOP) were checked pre- and postoperatively. RESULTS: Mean pain scores were 0.12 (maximum: 3) in group 1 and 0.37 in group 2; the difference between the groups was not statistically significant (95% confidence interval [CI] 0.003-0.487; p = 0.05). A total of 48 patients in group 1 (98%) and 49 in group 2 (100%) reported no discomfort or only mild discomfort. No significant differences in flare and cells in the anterior chamber or IOP were found between the two study groups. There was a significantly greater frequency of corneal oedema in group 1 (p = 0.001). Postoperative central corneal thickness values were also significantly higher in group 1 (95% CI 11.64-57.24; p = 0.003). CONCLUSIONS: Results suggest that viscoanaesthesia provides a level of comfort during cataract surgery under topical anaesthesia similar to that facilitated by intracameral lidocaine. However, patients who are given viscoanaesthesia may have increased risk for postoperative corneal oedema. PMID- 19302078 TI - Nd:YAG laser goniopuncture after deep sclerectomy: outcomes. AB - PURPOSE: This study aimed to investigate the safety and efficacy of Nd:YAG laser goniopuncture (LGP) in lowering intraocular pressure (IOP) after deep sclerectomy (DS). METHODS: We reviewed the outcomes in 258 eyes of 258 consecutive patients who underwent DS or combined phacoemulsification and DS between August 2001 and December 2003. Mitomycin C was used during surgery in 192 eyes (74.4%). RESULTS: Mean follow-up was 40 +/- 11 months after DS and 31 +/- 12 months after LGP. Laser goniopuncture was performed in 173 eyes (67.0%). The mean interval between LGP and DS was 10.3 +/- 8.7 months. The probability of LGP being performed was 63.1% (95% confidence interval [CI] 57.3-69.4) at 36 months after surgery. At 2 years after LGP, the probability of maintaining IOP < 15 mmHg with a 20% decrease from the pre-laser IOP and no further glaucoma procedure or medication was 49.7% (95% CI 41.9-57.1). Iris covering the trabeculo-Descemet's membrane was the only factor significantly associated with the failure of LGP (hazard ratio 3.0, p < 0.001). Complications observed after LGP included peripheral anterior synechiae in 23 (13.2%), hypotony in seven (4.0%), late acute IOP rise in three (1.7%), delayed bleb leak in one and blebitis in two eyes. Argon laser iridoplasty was performed in 45 (26.0%) and needle revision in 41 (23.7%) eyes. CONCLUSIONS: ND:YAG laser goniopuncture is an effective procedure to further lower IOP after DS. An IOP of < 15 mmHg is achieved and maintained for > or = 2 years in about 50% of cases after a single LGP procedure. There are potentially serious complications associated with the procedure. Regular gonioscopy is recommended following LGP to detect any occlusion of the goniopuncture by the iris. PMID- 19302079 TI - Descemet's stripping automated endothelial keratoplasty and penetrating keratoplasty for Fuchs' endothelial dystrophy. AB - PURPOSE: To compare the outcome of Descemet's stripping endothelial keratoplasty (DSAEK) to that of penetrating keratoplasty (PK) in patients with Fuchs' endothelial dystrophy. METHODS: The first 20 patients who underwent DSAEK at the Department of Ophthalmology, Aarhus University Hospital were compared to 20 patients treated with classic PK. Best-corrected visual acuity, subjective spectacle refraction and corneal thickness were registered before surgery and 1, 3, 6 and 12 months after DSAEK surgery; they were also measured before surgery and 12 months and 2-3 years after PK. Endothelial cell density was measured 12 months after surgery in both groups. RESULTS: Two primary graft failures were observed in the DSAEK group; no failures were seen in the PK group. Best spectacle-corrected visual acuity (BSCVA) at 12 months after surgery was significantly better in the DSAEK group (0.56 +/- 0.04) than in the PK group (0.33 +/- 0.06). At this time, 70% of the DSAEK-treated eyes but only 25% of PK treated eyes had obtained a BSCVA of 0.5 or better. Two to three years after surgery, BSCVA was 0.5 or better in 55% of PK-treated eyes. Refractive ametropia and astigmatism were significantly smaller in DSAEK-treated eyes than in PK treated eyes, even after suture removal and arcuate keratotomy. Endothelial cell density (cells/mm2) after 1 year was lower in DSAEK-treated (1.338 +/- 113) than in PK-treated eyes (1.610 +/- 124), but the difference was not statistically significant. CONCLUSION: DSAEK seems to be superior to PK in treating Fuchs' endothelial keratoplasty, although primary graft failure may be more common. Visual recovery is faster, and major ametropia and astigmatism is not induced. Long-term follow-up studies are essential to assess whether this conclusion also holds true more than 1 year after surgery. PMID- 19302080 TI - Macular oedema as a sign of preventable neurological disease in a 17-year-old man. PMID- 19302081 TI - Occurrence of human papillomavirus in pterygia. AB - PURPOSE: The aim of the study was to assess the occurrence of human papillomavirus (HPV) DNA in pterygium. METHODS: The study involved 89 patients undergoing surgical procedures at the Department of Ophthalmology, Medical University of Lublin, Poland. Group 1 included 58 patients with clinically diagnosed pterygium. Group 2 consisted of 31 individuals with normal conjunctiva. The material was collected during elective surgical procedures. The presence of HPV genome was determined using polymerase chain reaction (PCR). Once the presence of HPV DNA was confirmed, 28 HPV genotypes were determined using reverse hybridization. RESULTS: The determinations confirmed the presence of HPV DNA in pterygium. In the material collected from 58 cases of pterygium (group 1), HPV DNA was identified in 16 patients (27.6%). In the material from 31 diagnostic specimens of normal conjunctiva (group 2), the presence of HPV was demonstrated in three cases (9.7%). A statistically significant difference was found in the presence of HPV DNA between the patients from groups 1 and 2 (p = 0.041). HPV type 16 was most common and was demonstrated in 56% of HPV-positive cases of pterygium. HPV 16 and HPV 6 co-infections were found in 19% of cases, while HPV 18 and HPV 6 co-infections were observed in 13%. In group 2, all three patients with HPV showed HPV 18. CONCLUSION: It seems that HPV is not necessary to induce pterygium; however, it might play a synergistic role in the multi-stage process of its development. PMID- 19302082 TI - Ethanol-mediated fetal dysmorphology and its relationship to the ontogeny of maternal liver metallothionein. AB - BACKGROUND: Fetal zinc (Zn) deficiency arising from ethanol-induction of the Zn binding protein metallothionein (MT) in the mother's liver has been proposed as a mechanism of teratogenicity. Here, we determine the ontogeny of MT and Zn homeostasis in rats and mice and then examine the effect of acute ethanol exposure in early embryonic development on this relationship. The protective effect of Zn against ethanol-mediated fetal dysmorphology is also examined. METHODS: Study 1: Maternal liver MT and Zn homeostasis was determined in Sprague Dawley rats and C57BL/6J mice throughout gestation. Study 2: Rats were administered ethanol (25% in saline, intraperitoneal 0.015 ml/g) or vehicle alone on gestational day (GD) 9. Maternal liver MT and Zn, and plasma Zn was determined over the ensuing 24 hours. Study 3: Pregnant rats were treated with ethanol and Zn (s.c. 2.5 microg Zn/g) on GD9 and fetal dysmorphology was assessed on GD 19. RESULTS: Study 1: Maternal liver MT began to rise around GD 9 peaking on GD 15 before falling to nonpregnant levels around term. The pregnancy-related increase in MT was associated with a fall in plasma Zn which was significantly lower on GD 15 thereafter returning to nonpregnant levels by parturition. Study 2: Ethanol administered to pregnant rats on GD 9 resulted in a 10-fold induction of MT in the maternal liver and was associated with a 33% rise in liver Zn and a 30% fall in plasma Zn, 16 hours after treatment. Study 3: Ethanol treatment on GD 9 resulted in a significant increase in craniofacial malformations which were prevented by concurrent Zn treatment. CONCLUSIONS: The findings indicate that maternal liver MT levels are lowest in early gestation (before GD 10) making this a sensitive period where ethanol-induction of MT can affect fetal Zn homeostasis and cause fetal dysmorphology. The study further provides evidence of a protective role for Zn against ethanol-mediated teratogenicity. PMID- 19302083 TI - Naltrexone decreases heavy drinking rates in smoking cessation treatment: an exploratory study. AB - BACKGROUND: There is mixed support for the efficacy of the opioid antagonist naltrexone in the treatment of nicotine dependence. One potential unexplored mechanism underlying naltrexone's effects in smoking cessation may be in its ability to reduce alcohol consumption. METHODS: Alcohol consumption and liver enzyme levels (aspartate aminotransferase and alanine transaminase) were examined in a sample of 78 nonalcoholic social drinking smokers (34 naltrexone, 44 placebo) enrolled in a double-blind randomized clinical trial of naltrexone in smoking cessation. Naltrexone or placebo began 3 days prior to the quit date (25 mg daily) and continued for 8 weeks (50 mg daily). All participants received nicotine patches and behavioral counseling up through 4 weeks after the quit date. RESULTS: Naltrexone significantly reduced weekly heavy drinking rates. This effect was associated with greater nausea and pill taking adherence within the naltrexone group. Within heavy drinkers, naltrexone also directionally improved smoking quit rates compared with placebo. Liver enzyme levels did not differ during treatment with naltrexone compared with placebo. CONCLUSIONS: Naltrexone may reduce the frequency of heavy drinking in nonalcoholics attempting to quit smoking. Further, naltrexone may preferentially improve smoking quit rates within heavy drinkers who smoke, and further investigation in larger sample sizes is warranted. PMID- 19302084 TI - Effect of acute ethanol administration on the release of opioid peptides from the midbrain including the ventral tegmental area. AB - BACKGROUND: Experimental evidence suggests that ethanol alters the activity of the endogenous opioid peptide systems in a dose and brain-region dependent manner. These alterations may influence the processes of ethanol reward and reinforcement. Thus, it was the objective of this study to investigate the response of the 3 major opioid peptide systems (endorphins, enkephalins, and dynorphins) to acute ethanol administration, at the level of the midbrain including the ventral tegmental area (midbrain/VTA), a region important for drug, including ethanol reinforcement. METHODS: Using the in vivo microdialysis technique coupled with specific solid-phase radioimmunoassay for beta-endorphin, met-enkephalin, and dynorphin A(1-8,) changes in the extracellular concentration of theses peptides at the level of midbrain/VTA were determined at distinct time points following the administration of 0.0 (saline), 0.8, 1.2, 1.6, 2.0, and 2.4 g ethanol/kg B.Wt. RESULTS: A biphasic effect of ethanol on beta-endorphin release was found, with low to medium (1.2, 1.6, and 2.0 g) but not high (2.4 g) doses of ethanol, inducing a significant increase in the dialysate content of beta-endorphin. A late increase in the dialysate content of dynorphin A(1-8) was observed in response to the 1.2 g ethanol dose. However, none of the ethanol doses tested significantly altered the content of met-enkephalin in the dialysate. CONCLUSIONS: The present findings suggest that the ethanol-induced increase of beta-endorphin release at the level of midbrain/VTA may influence alcohol reinforcement. PMID- 19302085 TI - Alcohol consumption, social support, and risk of stroke and coronary heart disease among Japanese men: the JPHC Study. AB - BACKGROUND: It is unclear whether the association between alcohol consumption and risk of cardiovascular disease is affected by social support. METHODS: The prospective data for 19,356 men aged 40 to 69 years who participated in the Japan Public Health Center-Based Prospective Study. Alcohol consumption was classified into 7 categories: never, past, occasional, 1 to 149, 150 to 299, 300 to 449, or > or =450 g ethanol/wk. Associations between alcohol consumption and risk of cardiovascular disease were stratified by the median level of social support score, which was measured in emotional support score of this cohort study. RESULTS: During an average follow-up of 9.9 years, 629 total strokes and 207 coronary heart diseases were documented. Light-to-moderate alcohol consumption was associated with reduced risks of coronary heart disease and total cardiovascular disease, while heavy alcohol consumption was associated with increased risk of total stroke, in particular hemorrhagic stroke. When stratified by social support score, the multivariable hazard ratios of total cardiovascular disease associated with light-to-moderate alcohol consumption (1 to 299 g/wk) were 0.99 (0.72 to 1.37) in the low social support group and 0.56 (0.44 to 0.70) in the high social support group (p for interaction = 0.002), while the multivariable hazard ratios of hemorrhagic stroke associated with heavy alcohol consumption (> or =300 g/wk) were 2.09 (1.03 to 4.27) in the low social support group and 1.25 (0.72 to 2.15) in the high social support group (p for interaction = 0.44). There was no interaction between alcohol consumption and social support in relation to risk of coronary heart disease. CONCLUSIONS: Social support may enhance the beneficial effect of light-to-moderate alcohol consumption on risk of cardiovascular disease. PMID- 19302086 TI - GDNF is an endogenous negative regulator of ethanol-mediated reward and of ethanol consumption after a period of abstinence. AB - BACKGROUND: We previously found that activation of the glial cell line-derived neurotrophic factor (GDNF) pathway in the ventral tegmental area (VTA) reduces ethanol-drinking behaviors. In this study, we set out to assess the contribution of endogenous GDNF or its receptor GFRalpha1 to the regulation of ethanol-related behaviors. METHODS: GDNF and GFRalpha1 heterozygote mice (HET) and their wild type littermate controls (WT) were used for the studies. Ethanol-induced hyperlocomotion, sensitization, and conditioned place preference (CPP), as well as ethanol consumption before and after a period of abstinence were evaluated. Blood ethanol concentration (BEC) was also measured. RESULTS: We observed no differences between the GDNF HET and WT mice in the level of locomotor activity or in sensitization to ethanol-induced hyperlocomotion after systemic injection of a nonhypnotic dose of ethanol and in BEC. However, GDNF and GFRalpha1 mice exhibited increased place preference to ethanol as compared with their WT littermates. The levels of voluntary ethanol or quinine consumption were similar in the GDNF HET and WT mice, however, a small but significant increase in saccharin intake was observed in the GDNF HET mice. No changes were detected in voluntary ethanol, saccharin or quinine consumption of GFRalpha1 HET mice as compared with their WT littermates. Interestingly, however, both the GDNF and GFRalpha1 HET mice consumed much larger quantities of ethanol after a period of abstinence from ethanol as compared with their WT littermates. Furthermore, the increase in ethanol consumption after abstinence was found to be specific for ethanol as similar levels of saccharin intake were measured in the GDNF and GFRalpha1 HET and WT mice after abstinence. CONCLUSIONS: Our results suggest that endogenous GDNF negatively regulates the rewarding effect of ethanol and ethanol drinking behaviors after a period of abstinence. PMID- 19302087 TI - Magnetic resonance microscopy defines ethanol-induced brain abnormalities in prenatal mice: effects of acute insult on gestational day 8. AB - BACKGROUND: Magnetic resonance microscopy (MRM), magnetic resonance imaging (MRI) at microscopic levels, provides unprecedented opportunities to aid in defining the full spectrum of ethanol's insult to the developing brain. This is the first in a series of reports that, collectively, will provide an MRM-based atlas of developmental stage-dependent structural brain abnormalities in a Fetal Alcohol Spectrum Disorders (FASD) mouse model. The ethanol exposure time and developmental stage examined for this report is gestational day (GD) 8 in mice, when the embryos are at early neurulation stages; stages present in humans early in the fourth week postfertilization. METHODS: For this study, pregnant C57Bl/6J mice were administered an ethanol dosage of 2.8 g/kg intraperitoneally at 8 days, 0 hour and again at 8 days, 4 hours postfertilization. On GD 17, fetuses that were selected for MRM analyses were immersion fixed in a Bouin's/Prohance solution. Control fetuses from vehicle-treated dams were stage-matched to those that were ethanol-exposed. The fetal mice were scanned ex vivo at 7.0 T and 512 x 512 x 1024 image arrays were acquired using 3-D spin warp encoding. The resulting 29 microm (isotropic) resolution images were processed using ITK-SNAP, a 3-D segmentation/visualization tool. Linear and volume measurements were determined for selected brain, head, and body regions of each specimen. Comparisons were made between control and treated fetuses, with an emphasis on determining (dis)proportionate changes in specific brain regions. RESULTS: As compared with controls, the crown-rump lengths of stage-matched ethanol-exposed GD 17 fetuses were significantly reduced, as were brain and whole body volumes. Volume reductions were notable in every brain region examined, with the exception of the pituitary and septal region, and were accompanied by increased ventricular volumes. Disproportionate regional brain volume reductions were most marked on the right side and were significant for the olfactory bulb, hippocampus, and cerebellum; the latter being the most severely affected. Additionally, the septal region and the pituitary were disproportionately large. Linear measures were consistent with those of volume. Other dysmorphologic features noted in the MR scans were choanal stenosis and optic nerve coloboma. CONCLUSIONS: This study demonstrates that exposure to ethanol occurring in mice at stages corresponding to the human fourth week postfertilization results in structural brain abnormalities that are readily identifiable at fetal stages of development. In addition to illustrating the utility of MR microscopy for analysis of an FASD mouse model, this work provides new information that confirms and extends human clinical observations. It also provides a framework for comparison of structural brain abnormalities resulting from ethanol exposure at other developmental stages and dosages. PMID- 19302089 TI - MAOA-uVNTR polymorphism may modify the protective effect of ALDH2 gene against alcohol dependence in antisocial personality disorder. AB - BACKGROUND: Antisocial alcoholism is related to dopamine and serotonin which are catalyzed by monoamine oxidase A (MAOA) and acetaldehyde dehydrogenase 2 (ALDH2). The objective of this study is to determine whether the interaction between the MAOA and the ALDH2 genes is associated with subjects with antisocial personality disorder (ASPD) having alcoholism. METHODS: A total of 294 Han Chinese men in Taiwan including 132 ASPD with alcoholism (Antisocial ALC) and 162 without alcoholism (Antisocial Non-ALC) were recruited in this study. Alcohol dependence and ASPD were diagnosed according to DSM-IV criteria. Genotypes of ALDH2 and MAOA uVNTR were determined using PCR-RFLP. RESULTS: A significant difference of ALDH2 polymorphisms (p = 3.39E-05), but not of MAOA, was found among the 2 study groups. However, only after the stratification of the MAOA-uVNTR (variable number of tandem repeat located upstream) 3-repeat, a significant association between Antisocial Non-ALC and ALDH2*1/*2 or *2/*2 genotypes was shown (p = 1.46E-05; odds ratio = 3.913); whereas stratification of MAOA-uVNTR 4-repeat revealed no association. Multiple logistic regression analysis further revealed significant interaction of MAOA and ALDH2 gene in antisocial ALC (odds ratio = 2.927; p = 0.032). CONCLUSION: The possible interaction of MAOA and ALDH2 gene is associated with Antisocial ALC in Han Chinese males in Taiwan. However, the protective effects of the ALDH2*2 allele against alcoholism might disappear in subjects with ASPD and carrying MAOA-uVNTR 4-repeat allele in the Han Chinese male population. PMID- 19302088 TI - Ethanol-induced social facilitation in adolescent rats: role of endogenous activity at mu opioid receptors. AB - BACKGROUND: Ethanol consumption is considerably elevated during adolescence. Attractiveness of alcohol for humans during the adolescent developmental period is based, in part, on its ability to induce social facilitation--a facilitation of social interactions not only evident in human adolescents but also in adolescent rats. Endogenous opioid systems are among the multiple neural systems implicated in the behavioral and reinforcing effects of ethanol and may play a substantial role in modulating stimulatory effects of low doses of ethanol on social behavior during adolescence. This possibility was explored in the present study through the use of an animal model of peer-directed social behavior. METHODS: Sprague-Dawley rats were challenged early in adolescence with saline or ethanol intraperitoneally (i.p.), placed into an individual holding cage for 30 minutes, and then tested in a familiar situation with a nonmanipulated partner of the same age and sex. In Experiment 1, each test subject was injected subcutaneously with one of the three doses of a nonselective opioid antagonist naloxone (0, 0.05, and 0.1 mg/kg), 5 minutes prior to the social interaction test and 25 minutes following challenge with saline or ethanol (0.5 g/kg), whereas in Experiment 2 animals were challenged with one of the six doses of ethanol (0, 0.25, 0.5, 0.75, 1.0, and 1.25 g/kg) prior to injection of either saline or naloxone (0.05 mg/kg). In Experiment 3, animals were pretreated i.p. with the selective mu-opioid antagonist CTOP (0, 0.01, 0.025, 0.05, and 0.1 mg/kg) 30 minutes prior to challenge with saline or ethanol (0.5 g/kg). RESULTS: Low doses of ethanol (0.5 and 0.75 g/kg) produced social facilitation, as indexed by significant increases in play fighting and social investigation. Both doses of naloxone and the three highest doses of CTOP blocked the stimulatory effects of ethanol on play fighting but not on social investigation. These effects were not associated with alterations in ethanol pharmacokinetic properties or with shifts in the biphasic ethanol dose-response curve. CONCLUSIONS: Ethanol-induced facilitation of social play behavior seen in adolescent animals is mediated in part through ethanol-induced release of endogenous ligands for the mu-opioid receptor or an ethanol-associated enhancement of sensitivity of these receptors for their endogenous ligands. PMID- 19302090 TI - Screening trauma patients with the alcohol use disorders identification test and biomarkers of alcohol use. AB - BACKGROUND: Alcohol screening and brief interventions have been shown to reduce alcohol-related morbidity in injured patients. Use of self-report questionnaires such as the Alcohol Use Disorder Identification Test (AUDIT) is recommended as the optimum screening method. We hypothesized that the accuracy of screening is enhanced by combined use of the AUDIT and biomarkers of alcohol use in injured patients. METHODS: The study was conducted in the emergency department of a large, urban, university hospital. Patients were evaluated with the AUDIT, and blood sampled to determine carbohydrate-deficient transferrin, gamma-glutamyl transferase, and mean corpuscular volume. Alcohol problems were defined as presence of ICD-10 criteria for dependence or harmful use, or high-risk drinking according to World Health Organization criteria (weekly intake >420 g in males, >280 g in females). Screening accuracy was determined using Receiver Operating Characteristic curves. RESULTS: There were 787 males and 446 females in the study. Median age was 33 years. The accuracy of the AUDIT was good to excellent, whereas all biomarkers performed only fairly to poorly in males, and even worse in females. At a specificity >0.80, sensitivity for all biomarkers was <0.43, whereas sensitivity for the AUDIT was 0.76 for males and 0.81 for females. The addition of biomarkers added little additional discriminatory information compared to use of the AUDIT alone. CONCLUSIONS: Screening properties of the AUDIT are superior to %CDT, MCV, and GGT for detection of alcohol problems in injured patients and are not clinically significantly enhanced by the use of biomarkers. PMID- 19302091 TI - Ethanol-sensitive brain regions in rat and mouse: a cartographic review, using immediate early gene expression. AB - BACKGROUND: Ethanol addiction has been conceptualized as a progression from occasional, impulsive use to compulsive behavior. Ethanol-dependence is a chronic pathology with repeated cycles of withdrawal, craving, and relapse. Specific molecular and cellular mechanisms underlie these transition stages. METHODS: This review aimed at elucidating whether there are also adaptations in the pattern of brain regions responding to ethanol. This paper reviews the evidence in rodents for activation of specific brain regions, assessed by induction of IEG expression, following acute and chronic ethanol exposure. RESULTS: The review sheds light on the specific patterns of response in regions of the brain to different types of ethanol exposure and shows that activation of specific brain regions may occur in particular phases of the development of ethanol addiction. Some brain regions respond consistently following acute or chronic treatments or withdrawal: the prefrontal cortex; nucleus accumbens; lateral septum; hippocampus; perioculomotor urocortin-containing cells population (pIIIu), also known as Edinger-Westphal nucleus; central nucleus of the amygdala; and the paraventricular nucleus of hypothalamus. The two last brain areas are particularly activated by relapse-inducing stressors. It is of interest that the amygdala, hippocampus, and prefrontal cortex, which belong to the reward system, are activated by cue-induced relapse to ethanol self-administration in rodents and humans, while activation of these regions is reversed with anti-craving compounds. Following chronic exposure, IEG induction desensitizes while withdrawal reactivates these regions. DISCUSSION: Some responding regions are implicated in reward related processes (VTA, extended amygdala, hypothalamus, hippocampus, prelimbic cortex, ventral part of lateral septum) and some others in aversive-related processes (area postrema, nucleus of solitary tract). CONCLUSION: A better understanding of the neural circuits affected by ethanol and their adaptations during the development of ethanol addiction will provide new opportunities for developing appropriate therapies. PMID- 19302092 TI - Severity of perinatal illness and cerebral cortical growth in preterm infants. AB - AIM: We have shown previously that the degree of prematurity affects cortical surface area growth. We now addressed the question whether cortical surface area growth after preterm birth is predicted by the severity of peri- and postnatal illness. METHODS: Cortical surface area was measured in 269 images from 111 infants born between 23 and 29 weeks and imaged at 23 to 48 weeks gestational age (GA). The severity of perinatal illness was assessed using the clinical risk index for babies score (CRIB I) and the severity of ongoing illness by the presence of chronic lung disease (CLD). The effects on cortical growth were modelled using generalized least-square regression for random effects with Bonferroni correction. To explore the results further we examined CRIB II, C reactive protein (CRP) on the second day after birth, and time taken to achieve full enteral feeding. RESULTS: Cortical surface area grew by 12.4% per week. Reduced cortical growth was predicted by adverse CRIB I (-0.15% per week per unit) and development of CLD (-1.18% per week). Secondary analysis showed that growth was related to adverse CRIB II (-0.36% per week per unit) and increasing CRP (-0.03% per week per mMol), but not by the time taken to achieve full enteral feeding. CONCLUSION: After very premature birth illness severity predicts reduced cortical growth. PMID- 19302093 TI - Environmental tobacco smoke exposure as a risk factor for infections in infancy. AB - AIM: This prospective observational study investigated the effect of environmental tobacco smoke (ETS) on frequency and severity of common infantile infections. METHODS: In a representative sample of 926 infants, parental smoking was recorded at months 1 and 9 postpartum, and all infantile infectious episodes were recorded at 1, 3, 6, 9 and 12 months postpartum. RESULTS: Both parents were regular smokers all through the first year in 107 (11.6%), at least one smoked regularly or occasionally in 492 (53.1%), and parents did not smoke at all in 327 (35.3%) families. Among mothers, 168 (18.1%) smoked perinatally. Infantile ETS exposure was associated with increased frequency of total infectious episodes (p = 0.025) and hospitalizations for infection (p = 0.007). In ETS exposed infants, birth in autumn and presence of siblings contributed to increased frequency of most infections and of hospital admissions for infection. By contrast, exclusive breastfeeding protected against the effect of ETS on total infantile infections (OR 0.982, 95% CI 0.965-0.999; p = 0.036), hospital admissions for infection (OR 0.980, 95% CI 0.961-0.999; p = 0.036) and thrush (OR 0.973, 95% CI 0.951-0.996; p = 0.022). CONCLUSION: Our findings point to harmful effect of ETS on infantile health and further suggest that this effect may be enhanced or diminished by other factors. ETS should be regarded as a preventable risk factor for infections in infancy. PMID- 19302094 TI - Inbreeding variability and population structure in the invasive haplodiploid palm seed borer (Coccotrypes dactyliperda). AB - We investigated the mating system and population genetic structure of the invasive haplodiploid palm-seed borer Coccotrypes dactyliperda in California. We focused on whether these primarily inbreeding beetles have a 'mixed-breeding' system that includes occasional outbreeding, and whether local inbreeding coefficients (F(IS)) varied with dominant environmental factors. We also analysed the genetic structure of C. dactyliperda populations across local and regional scales. Based on the analysis of genetic variation at seven microsatellite loci in 1034 individual beetles from 59 populations, we found both high rates of inbreeding and plentiful evidence of mixed-breeding. F(IS) ranged from -0.56 to 0.90, the highest variability reported within any animal species. There was a negative correlation between F(IS) and latitude, suggesting that some latitude associated factor affecting mating decisions influenced inbreeding rates. Multiple regressions suggested that precipitation, but not temperature, may be an important correlate. Finally, we found highly significant genetic differentiation among sites, even over short geographic distances (< 1000 m). PMID- 19302095 TI - Association of Mycobacterium tuberculosis complex isolates of BOVIS and Central Asian (CAS) genotypic lineages with extrapulmonary disease. AB - The association between isolate genotype, defined as in the international spoligotype database SpolDB4, and extrapulmonary tuberculosis was determined among 1009 patients in a population-based, 4-year survey performed in Tuscany, Italy. Extrapulmonary disease occurred in 24.2% of patients. A statistically significant association with extrapulmonary disease was found for the BOVIS (adjusted OR 3.2; 95% CI 1.2-8.1) and for the Central Asian (CAS) lineages (adjusted OR 2.3; 95% CI 1.0-5.1). These findings support the view that Mycobacterium tuberculosis strains within individual genotypic lineages might have evolved unique pathogenic characteristics that are capable of influencing the clinical outcome of the infection. PMID- 19302096 TI - Oxidative stress sensitivity in Debaryomyces hansenii. AB - Debaryomyces hansenii is an osmotolerant and halotolerant yeast of increasing interest for fundamental and applied research. In this work, we have performed a first study on the effect of oxidative stress on the performance of this yeast. We have used Saccharomyces cerevisiae as a well-known reference yeast. We show that D. hansenii is much more susceptible than S. cerevisiae to cadmium chloride, hydrogen peroxide or 1,4-dithiothreitol. These substances induced the formation of reactive oxygen species (ROS) in both yeasts, the amounts measured being significantly higher in the case of D. hansenii. We also show that NaCl exerted a protective effect against oxidative stress in Debaryomyces, but that this was not the case in Saccharomyces because sodium protected that yeast only when toxicity was induced with cadmium. On the basis of the present results, we raised the hypothesis that the sensitivity to oxidative stress in D. hansenii is related to the high amounts of ROS formed in that yeast and that observations such as low glutathione amounts, low basal superoxide dismutase and peroxidase activities, decrease in ATP levels produced in the presence of ROS inducers and high cadmium accumulation are determinants directly or indirectly involved in the sensitivity process. PMID- 19302097 TI - Methylobacterium sp. resides in unculturable state in potato tissues in vitro and becomes culturable after induction by Pseudomonas fluorescens IMGB163. AB - AIMS: To induce growth of endophytic bacteria residing in an unculturable state in tissues of in vitro-grown potato plantlets. To isolate and identify the induced bacteria and to localize the strains in tissues of in vitro-grown potato plantlets. METHODS AND RESULTS: The inoculation of in vitro-grown potato plants with Pseudomonas fluorescens IMBG163 led to induction of another bacterium, a pink-pigmented facultative methylotroph that was identified as Methylobacterium sp. using phylogenetic 16S rDNA approach. Two molecular methods were used for localizing methylobacteria in potato plantlets: PCR and in situ hybridization (ISH/FISH). A PCR product specific for the Methylobacterium genus was found in DNA isolated from the surface-sterilized plantlet leaves. Presence of Methylobacterium rRNA was detected by ISH/FISH in leaves and stems of inoculated as well as axenic potato plantlets although the bacterium cannot be isolated from the axenic plants. CONCLUSION: Methylobacterium sp. resides in unculturable state within tissues of in vitro-grown potato plants and becomes culturable after inoculation with P. fluorescens IMBG163. SIGNIFICANCE AND IMPACT OF THE STUDY: In order to develop endophytic biofertilizers and biocontrol agents, a detailed knowledge of the life-style of endophytes is essential. To our knowledge, this is the first report on increase of the culturability of endophytes in response to inoculation by nonpathogenic bacteria. PMID- 19302098 TI - Novel approach to control Salmonella enterica by modern biophotonic technology: photosensitization. AB - AIMS: Salmonellosis is one of the most common foodborne diseases in the world. The aim of this study was to evaluate the antibacterial efficiency of 5 aminolevulinic acid (ALA) based photosensitization against one of food pathogens Salmonella enterica. METHODS AND RESULTS: Salmonella enterica was incubated with ALA (7.5 mmol l(-1)) for 1-4 h and afterwards illuminated with visible light. The light source used for illumination of S. enterica emitted light lambda = 400 nm with energy density 20 mW cm(-2). The illumination time varied from 0 to 20 min and subsequently a total energy dose reached 0-24 J cm(-2). The data obtained indicate that S. enterica is able to produce endogenous photosensitizer PpIX when incubated with ALA. Remarkable inactivation of micro-organisms can be achieved (6 log) after photosensitization. It is obvious that photosensitization-based inactivation of S. enterica depends on illumination as well as incubation with ALA time. CONCLUSION: ALA-based photosensitization can be an effective tool against multi-drug resistant Gram-negative bacteria S. enterica serovar Typhimurium. SIGNIFICANCE AND IMPACT OF THE STUDY: Experimental data and mathematical evaluations support the idea that ALA-based photosensitization can be a useful tool for the development of nonthermal food preservation technology in future. PMID- 19302099 TI - Proteomic analysis of lactose-starved Lactobacillus casei during stationary growth phase. AB - AIMS: Starvation stress is a condition that nonstarter lactic acid bacteria (NSLAB) normally encounter. This study was aimed to investigate starvation induced proteins in Lactobacillus casei during stationary growth phase. METHODS AND RESULTS: The impact of carbohydrate starvation on L. casei GCRL163 was investigated using two different media (a modified de Man, Rogosa and Sharpe broth and a semi-defined medium). Cells were grown in the presence of excess lactose (1%) or starvation (0%) and differences in the patterns of one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and two dimensional electrophoresis of the cytosolic protein fractions were investigated. Differentially regulated proteins were identified by MALDI-TOF/TOF mass spectrometry. Many differentially regulated proteins were enzymes of various metabolic pathways involved in carbohydrate metabolism to yield energy. Differences in protein expression were also observed in the two culture conditions tested in this experiment. CONCLUSION: Numerous glycolytic enzymes were differentially regulated under lactose starvation. The differential expression of these glycolytic enzymes suggests a potential survival strategy under harsh growth conditions (i.e. lactose starvation). SIGNIFICANCE AND IMPACT OF THE STUDY: This paper reports improved understanding of stress responses and survival mechanism of NSLAB under lactose-depleted cheese-ripening condition. This knowledge of how NSLAB bacteria adapt to lactose starvation could be applied to predict the performances of bacteria in other industrial applications. PMID- 19302100 TI - Peptidases specific for proline-containing peptides and their unusual peptide dependent regulation in Oenococcus oeni. AB - AIMS: Growth of the lactic acid bacterium (LAB) Oenococcus oeni, which is involved in malolactic fermentation during the winemaking process, is stimulated by peptides originating from yeast. In this study, we investigated the impact of peptides on O. oeni growth, peptidase activity and the expression of genes encoding the studied peptidases. METHODS AND RESULTS: Low levels of PepN activity and very high levels of PepI activity were observed in O. oeni, whereas levels of PepX activity were intermediate. The level of biosynthesis of these O. oeni peptidases was shown to depend on peptides present in the culture medium. These results were confirmed by transcriptional analyses of putative pep genes. The mechanism of repression by peptides did not involve a CodY-like regulator. CONCLUSIONS: Peptides from yeast decrease the levels of enzymatic activity and relative gene expression of O. oeni peptidases. Peptidases specific for proline containing peptides are important for O. oeni nitrogen metabolism. SIGNIFICANCE AND IMPACT OF THE STUDY: We report here for the first time that the enzymes involved in the assimilation of proline-containing peptides by O. oeni differ from the well-described proteolytic system of milk LAB. This may reflect a specific adaptation to the wine environment. PMID- 19302101 TI - Chelated iron as an anti-Pseudomonas aeruginosa biofilm therapeutic strategy. PMID- 19302102 TI - First fixed drug eruption due to teicoplanin with a peri-oral distribution. PMID- 19302103 TI - Hypertrophic pyloric stenosis and pulmonary hypertension in a neonate. A common mechanism? AB - Nitric oxide (NO) is an important mediator of biological functions. Absence or shortage of NO plays a role in the pathogenesis of both hypertrophic pyloric stenosis and persistent pulmonary hypertension. We present a neonate diagnosed with pulmonary hypertension after birth caused by meconium-aspiration syndrome eventually treated with extracorporal membrane oxygenation followed by hypertrophic pyloric stenosis for which a pyloromyotomy was performed. In conclusion, the association of pulmonary hypertension and pyloric stenosis has not been described before and may be explained by a lowered plasma concentration of arginine leading to deficient NO synthesis in the affected organ systems. PMID- 19302104 TI - Integrating hormones into the floral-transition pathway of Arabidopsis thaliana. AB - The transition from vegetative to reproductive growth is a major phase change in angiosperms. In annual plants such as Arabidopsis thaliana (Arabidopsis), this change is irreversible, and as such, the regulation of its timing must be tightly controlled. Plant hormone (phytohormone) signalling is known to regulate suites of morphogenic processes in Arabidopsis a role in flowering-time control is starting to emerge as one key-controlling step. This review focuses on experimental evidence in the Arabidopsis that both classical and newly described phytohormones serve within the signal network leading to a reproductive phase transition, as both positive and repressive elements, depending on the phytohormone and growth conditions. Examples of genetic and pharmacological experiments that implicate phytohormones as components of the floral-timing syndrome will be described. I hope that this review will serve as a primer for future research on the mechanisms of action for each respective phytohormone on the floral transition in Arabidopsis, and lead to further experimentation on the crosstalk that likely bridges between them. PMID- 19302105 TI - Evidence-based urology in practice: how to use PubMed effectively. PMID- 19302106 TI - Optimization of conjunctival biopsy specimen processing: a novel technique. PMID- 19302107 TI - Eating thyself toward the dark side? PMID- 19302113 TI - B cells and rejection: is there a link? Meaning and management of rejection characterized by steroid-resistance and B-cell infiltrates. PMID- 19302114 TI - The social aspects of food and nutrition. PMID- 19302115 TI - A history and review of school meal standards in the UK. AB - This review details the history of school meals in the UK, from their origin in the mid-19th Century, to the present day, and provides a summary of how each country has independently developed its own food and or nutrient-based standards for school meals. The standards in place in the UK are amongst some of the most detailed and comprehensive in the world. Regular monitoring to ensure that these standards are being met and that schools are improving healthy eating is essential to their success. Of no lesser importance are assessments to determine whether changes to school meals are having an impact on the diets of school children. It is early days in terms of evaluation because food-based standards have only recently been introduced and nutrient-based standards are in the process of being developed and implemented. Studies in England provide some evidence that the re-introduction of standards for school food is having a positive impact on both pupil's food choices and the nutritional profile of school lunches. At present, there does not appear to be a pattern between current obesity levels and the types of school meals provided, although it is anticipated that, in the long term, these comprehensive standards may contribute to a less obesogenic environment. PMID- 19302116 TI - A qualitative study of the food-related experiences of rural village shop customers. AB - BACKGROUND: In the UK, although food choice in urban contexts has been widely studied, far less empirical information has been gathered from rural settings. With the closure of local services, some rural dwellers were believed to experience difficulties in meeting healthy eating recommendations. The present study aimed to explore perceptions of village store users. METHODS: Semi structured, in-depth interviews were conducted individually with 40 adults who were frequent users of rural village shops in Norfolk, UK. Participants were purposively selected to ensure that a wide demographic cross-section of customers was obtained. Interviews focused upon food choice strategies; attitudes towards rural food retail; and the provision of healthy foods. Interviews were transcribed verbatim and analysed qualitatively, using an established interpretative phenomenological approach. RESULTS: Four main themes emerged as drivers of food choice in rural villages: (1) village store as icon, which described how the perceived centrality of village shops influenced food choices; (2) village store as a service provider, which described top-up shopping behaviours; (3) alternative food sources, which described store users desires for local foods and their attitudinal conflicts towards supermarkets; and (4) lifestyle factors, which described the influence of factors such as time pressures, access to cars and family structures. CONCLUSIONS: Food choices were strongly influenced by the distinctive characteristics of the rural environment. Village shops were seen as important for community identity ('rural idyll'), as well as providing access to food and services. However, desires were made apparent for a greater range of healthy, fresh and locally-sourced foods. PMID- 19302117 TI - Dietary beliefs and behaviour of a UK Somali population. AB - BACKGROUND: Somalis comprise one of the largest asylum seeking populations in the UK, yet very little is known about how this migration has affected traditional attitudes towards food or eating habits. The present study was commissioned to examine the health behaviours (smoking, diet and exercise) of a Somali population in London; the study focuses on the dietary beliefs and self-reported eating behaviours of these subjects. METHODS: Because of the documented difficulty of engaging black and minority ethnic groups in research, a mixed methodology was used, involving focus groups to elicit specific dietary themes and a questionnaire survey to attempt to quantify community concordance with these themes. Eight focus groups were held in London in 2006; there were two women's and six men's groups, with 62 participants in total. The questionnaire was developed following analysis of the focus group discussions and opportunistically distributed via local Somali community organisations (77 respondents). RESULTS: The typical diet of focus group participants largely consisted of rice, pasta and red meat. There was low consumption of fruit and vegetables reported among the focus group participants: of survey respondents 97% reported eating less than two pieces of fruit, and 92% less than two portions of vegetables, a day. CONCLUSIONS: Fruit and vegetable consumption was low and there was uncertainty about what constituted a healthy diet and a stated desire for education around this. Cultural factors such as the traditional Somali diet, social associations of food and lack of appropriate information are issues that need to be addressed. PMID- 19302118 TI - Psychosocial factors underlying the mother's decision to continue exclusive breastfeeding for 6 months: an elicitation study. AB - BACKGROUND: Despite numerous benefits of exclusive breastfeeding (EBF) for infants and mothers, a significant decrease in the EBF rate in the USA at six months compared to the rate at birth suggests that reasons for initiation and continuation of EBF may differ. The purpose of this qualitative study was to explore psychosocial factors underlying the continued EBF behaviour for six months, in order to identify salient belief structures according to the theory of planned behaviour. METHODS: Participants were recruited from central Indiana in the USA. They were asked to respond to an open-ended questionnaire designed to elicit positive/negative consequences, approving/disapproving social referents, and easy/difficult circumstances in continuing EBF for six months. Responses were translated into behavioural, normative, and control beliefs of the theory. RESULTS: Findings suggest that respondents (1) value emotional and health benefits of continued EBF for six months; (2) feel the approval from family and friends but disapproval from the society; (3) view health professional's position as positive and negative. CONCLUSIONS: Breastfeeding educators can more likely improve the EBF duration by addressing these salient beliefs. Identified beliefs provide a basis for the development of a quantitative instrument to further study the EBF behaviour. PMID- 19302119 TI - Effect of calorie restriction on subjective ratings of appetite. AB - BACKGROUND: Energy or calorie restriction (CR) has consistently been shown to produce weight loss and have beneficial health effects in numerous species, including primates and humans. Most individuals, however, are unable to sustain weight losses induced through reductions in energy intake, potentially due to increased hunger levels. The effects that prolonged CR has on subjective aspects of appetite have not been well studied. Thus, the present study tested the effect of 6 months of caloric restriction on appetite in healthy, overweight men and women. METHODS: Forty-eight overweight men and women with a body mass index (BMI; kg m(-2)) between 25-29.9 took part in a 6-month study and were randomised into one of four groups: healthy diet (control); 25% CR; 12.5% CR plus exercise (12.5% increased energy expenditure; CR + EX); low-calorie diet [LCD; 3724 kJ day(-1) (890 kcal day(-1)) until 15% of initial body weight was lost, then maintenance]. Appetite markers (i.e. hunger, fullness, desire to eat, etc.) were assessed weekly during a fasting state. RESULTS: Body weight was significantly reduced in all three energy-restricted groups (CR = -10.4 +/- 0.9%; CR + EX = -10.0 +/- 0.8%; and LCD = -13.9 +/-0.7%), indicating that participants were adherent to their energy restriction regimen, whereas the healthy diet control group remained weight stable (control = -1.0 +/- 1.1%). Despite these significant weight losses, appetite ratings of participants in the three energy-restricted groups at month 6 were similar to the weight stable control group. CONCLUSIONS: CR regimens with low fat diets producing significant weight losses have similar effects on appetite markers over a 6-month time period compared to a weight stable control group. PMID- 19302120 TI - Contribution of thickened drinks, food and enteral and parenteral fluids to fluid intake in hospitalised patients with dysphagia. AB - BACKGROUND: Studies amongst older people with acute dysphagic stroke requiring thickened fluids have assessed fluid intakes from combinations of beverage, food, enteral and parenteral sources, but not all sources simultaneously. The present study aimed to comprehensively assess total water intake from food, beverages, enteral and parenteral sources amongst dysphagic adult in-patients receiving thickened fluids. METHODS: Patients requiring thickened fluid following dysphagia diagnosis were recruited consecutively from a tertiary teaching hospital's medical and neurosurgical wards. Fluid intake from food and beverages was assessed by wastage, direct observation and quantified from enteral and parenteral sources through clinical medical records. RESULTS: No patients achieved their calculated fluid requirements unless enteral or parenteral fluids were received. The mean daily fluid intake from food was greater than from beverages whether receiving diet alone (food: 807 +/- 363 mL, food and beverages: 370 +/- 179 mL; P < 0.001) or diet with enteral or parenteral fluid support (food: 455 +/- 408 mL, food and beverages: 263 +/- 232 mL; P < 0.001). Greater daily fluid intakes occurred when receiving enteral and parenteral fluid in addition to oral dietary intake, irrespective of age group, whether assistance was required, diagnosis and whether stage 3 or stage 2 thickened fluids were required (P < 0.05). After enteral and parenteral sources, food provided the most important contribution to daily fluid intakes. CONCLUSIONS: The greatest contribution to oral fluid intake was from food, not beverages. Designing menus and food services that promote and encourage the enjoyment of fluid dense foods, in contrast to thickened beverages, may present an important way to improve fluid intakes of those with dysphagia. Supplemental enteral or parenteral fluid may be necessary to achieve minimum calculated fluid requirements. PMID- 19302124 TI - High and low, fast or slow: the complementary contributions of altitude and latitude to understand life-history variation. AB - Dark-eyed juncos (Junco hyemalis) from two elevations in the Rocky Mountains of Canada display different life histories. Birds breeding at high elevation face a later and compressed reproductive season during which they raise on average half the number of broods (young) than their low-elevation conspecifics. Fledglings at high elevation are heavier, fatter, and have an increased chance of surviving to 25 days of age. Likewise, survival of adults (males) increases at higher elevation. Put into a broader perspective, high-elevation juncos show the life history strategy characteristic for low-latitude birds. This raises questions about the mechanisms influencing life-history evolution. PMID- 19302125 TI - Trading offspring size for number in a variable environment: selection on reproductive investment in female Soay sheep. AB - 1. Given energetic constraints, female reproductive strategy is expected to be shaped by a trade-off between offspring size and offspring number, the optimal resolution of which may vary with environmental conditions. 2. We tested the hypothesis that selection will favour the production of larger litters, even at some expense to offspring size, under good conditions (and vice versa in harsh environments) using data from a long-term study of an unmanaged population of Soay sheep on the islands of St Kilda, NW Scotland. 3. Both litter size (which is either 1 or 2) and offspring birth weight are under positive selection through female annual fitness, but the strength of selection varies systematically with environmental conditions. Age effects were also detected, with selection weakening as female age increases. 4. Consistent with theoretical predictions, the selective trade-off between litter and offspring size is shifted in favour of the latter under poor environmental conditions. Thus, direct selection on offspring birth weight increases under harsh environmental conditions, particularly for females producing twins. 5. However, singletons are only favoured when environmental conditions are very poor, and offspring weight is constrained to be low. Thus, the current low incidence of twinning (13.5% of litters produced since 1985) appears to be suboptimal with respect to the empirically estimated selection regime. Thus, litter size, a trait known to be heritable, may be expected to evolve upwards in this population. 6. Our study highlights the necessity of incorporating environmental heterogeneity and age structure into analyses of natural selection, and suggests that the common assumption of optimality used in models of life history may sometimes be problematic. PMID- 19302126 TI - Clutch size determination in shorebirds: revisiting incubation limitation in the pied avocet (Recurvirostra avosetta). AB - 1. Traits strongly related to fitness, such as offspring number, are expected to show intraspecific variation among individuals. However, offspring number is invariant in several reptiles, birds, and mammals. Most shorebirds (210+ species), for example, have an invariant clutch size of four eggs, which is unexpected in such an ecologically, behaviourally and socially diverse group. 2. The incubation-limitation hypothesis (ILH) suggests that shorebird clutch size is limited by the inability of adults to incubate clutches larger than four eggs. Several recent studies reported no overall costs of incubating experimentally enlarged clutches and concluded no support for the traditional ILH. However, most studies have not measured all potential costs, and none has quantified costs beyond egg hatching. We conducted a clutch-enlargement experiment and measured potential costs both during incubation and chick rearing in pied avocets (Recurvirostra avosetta L.). 3. Hatching was more asynchronous and egg hatchability was marginally lower in enlarged clutches than in controls. Nonetheless, more young hatched from enlarged clutches (mean: 4.2 +/- 0.17 SE) than from controls (3.4 +/- 0.09), and the two groups did not differ in incubation period, complete or partial clutch failure, or hatchling body size, apparently refuting the ILH. 4. However, pairs incubating enlarged clutches occupied poorer feeding territories during chick rearing, experienced higher chick mortality, and eventually raised fewer young to independence (mean adjusted for season: 0.7 +/- 0.16 SE juveniles) than did control pairs (1.2 +/- 0.13). Chick survival was primarily associated with prey availability, and predation risks were not higher in larger broods. 5. Our results provide evidence that incubating unusually large clutches can affect post-hatching performance and lead to lower annual reproductive success in shorebirds. This study, therefore, supports the ILH and points to the importance of monitoring reproductive success beyond the hatching of the chicks. PMID- 19302127 TI - Spatial and temporal variation in the relative contribution of density dependence, climate variation and migration to fluctuations in the size of great tit populations. AB - 1. The aim of the present study is to model the stochastic variation in the size of five populations of great tit Parus major in the Netherlands, using a combination of individual-based demographic data and time series of population fluctuations. We will examine relative contribution of density-dependent effects, and variation in climate and winter food on local dynamics as well as on number of immigrants. 2. Annual changes in population size were strongly affected by temporal variation in number of recruits produced locally as well as by the number of immigrants. The number of individuals recruited from one breeding season to the next was mainly determined by the population size in year t, the beech crop index (BCI) in year t and the temperature during March-April in year t. The number of immigrating females in year t + 1 was also explained by the number of females present in the population in year t, the BCI in autumn year t and the temperature during April-May in year t. 3. By comparing predictions of the population model with the recorded number of females, the simultaneous modelling of local recruitment and immigration explained a large proportion of the annual variation in recorded population growth rates. 4. Environmental stochasticity especially caused by spring temperature and BCI did in general contribute more to annual fluctuations in population size than density-dependent effects. Similar effects of climate on local recruitment and immigration also caused covariation in temporal fluctuations of immigration and local production of recruits. 5. The effects of various variables in explaining fluctuations in population size were not independent, and the combined effect of the variables were generally non-additive. Thus, the effects of variables causing fluctuations in population size should not be considered separately because the total effect will be influenced by covariances among the explanatory variables. 6. Our results show that fluctuations in the environment affect local recruitment as well as annual fluctuations in the number of immigrants. This effect of environment on the interchange of individuals among populations is important for predicting effects of global climate change on the pattern of population fluctuations. PMID- 19302128 TI - From mechanistic insights to therapeutic applications. PMID- 19302129 TI - Snail puts melanoma on the fast track. PMID- 19302130 TI - Lipid-mediated signalling and melanocyte function. PMID- 19302131 TI - Highlights from the perspectives in melanoma XII conference. AB - The following meeting highlights are from the Perspectives in Melanoma XII conference, held in Scheveningen/The Hague, the Netherlands, on October 2-4, 2008. Selected reviews are included but further discussions of these and other presentations are posted at http://www.MelanomaCare.org. PMID- 19302132 TI - A Conversation with Anthony J. Schaeffer. Interview by Darracott Vaughan. PMID- 19302133 TI - Prostate cancer: a serious disease suitable for prevention. AB - Prostate cancer is among the most common causes of death from cancer in men, and accounts for 10% of all new male cancers worldwide. The diagnosis and treatment of prostate cancer place a substantial physical and emotional burden on patients and their families, and have considerable financial implications for healthcare providers and society. Given that the risk of prostate cancer continues to increase with age, the burden of the disease is likely to increase in line with population life-expectancy. Reducing the risk of prostate cancer has gained increasing coverage in recent years, with proof of principle shown in the Prostate Cancer Prevention Trial with the type 2 5alpha-reductase (5AR) inhibitor, finasteride. The long latency period, high disease prevalence, and significant associated morbidity and mortality make prostate cancer a suitable target for a risk-reduction approach. Several agents are under investigation for reducing the risk of prostate cancer, including selenium/vitamin E and selective oestrogen receptors modulators (e.g. toremifene). In addition, the Reduction by Dutasteride of Prostate Cancer Events trial, involving >8000 men, is evaluating the effect of the dual 5AR inhibitor, dutasteride, on the risk of developing prostate cancer. A successful risk-reduction strategy might decrease the incidence of the disease, as well as the anxiety, cost and morbidity associated with its diagnosis and treatment. PMID- 19302134 TI - Success or failure in failure? PMID- 19302136 TI - Nitrofurantoin: the return of an old friend in the wake of growing resistance. PMID- 19302137 TI - Robotic-assisted laparoscopic partial nephrectomy: surgical technique and clinical outcomes at 1 year. PMID- 19302138 TI - Laparoscopic right donor nephrectomy. PMID- 19302139 TI - Common themes emerge in the transcriptional control of T helper and developmental cell fate decisions regulated by the T-box, GATA and ROR families. AB - Cellular differentiation requires the precise action of lineage-determining transcription factors. In the immune system, CD4(+) T helper cells differentiate into at least three distinct effector lineages, T helper type 1 (Th1), Th2 and Th17, with the fate of the cell at least in part determined by the transcription factors T-box expressed in T cells (T-bet), GATA-3 and retinoid-related orphan receptor gammat (RORgammat), respectively. Importantly, these transcription factors are members of larger families that are required for numerous developmental transitions from early embryogenesis into adulthood. Mutations in members of these transcription factor families are associated with a number of human genetic diseases due to a failure in completing lineage-specification events when the factor is dysregulated. Mechanistically, there are both common and distinct functional activities that are utilized by T-box, GATA and ROR family members to globally alter the cellular gene expression profiles at specific cell fate decision checkpoints. Therefore, understanding the molecular events that contribute to the ability of T-bet, GATA-3 and RORgammat to define T helper cell lineages can provide valuable information relevant to the establishment of other developmental systems and, conversely, information from diverse developmental systems may provide unexpected insights into the molecular mechanisms utilized in T helper cell differentiation. PMID- 19302140 TI - Restricting activation-induced cytidine deaminase tumorigenic activity in B lymphocytes. AB - DNA breaks play an essential role in germinal centre B cells as intermediates to immunoglobulin class switching, a recombination process initiated by activation induced cytidine deaminase (AID). Immunoglobulin gene hypermutation is likewise catalysed by AID but is believed to occur via single-strand DNA breaks. When improperly repaired, AID-mediated lesions can promote chromosomal translocations (CTs) that juxtapose the immunoglobulin loci to heterologous genomic sites, including oncogenes. Two of the most studied translocations are the t(8;14) and T(12;15), which deregulate cMyc in human Burkitt's lymphomas and mouse plasmacytomas, respectively. While a complete understanding of the aetiology of such translocations is lacking, recent studies using diverse mouse models have shed light on two important issues: (1) the extent to which non-specific or AID mediated DNA lesions promote CTs, and (2) the safeguard mechanisms that B cells employ to prevent AID tumorigenic activity. Here we review these advances and discuss the usage of pristane-induced mouse plasmacytomas as a tool to investigate the origin of Igh-cMyc translocations and B-cell tumorigenesis. PMID- 19302141 TI - Oestrogen modulates experimental autoimmune encephalomyelitis and interleukin-17 production via programmed death 1. AB - The mechanism by which oestrogens suppress experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, is only partially understood. We here demonstrate that treatment with 17beta-oestradiol (E(2)) in C57BL/6 mice boosted the expression of programmed death 1 (PD-1), a negative regulator of immune responses, in the CD4(+) FoxP3(+) regulatory T (Treg) cell compartment in a dose-dependent manner that correlated with the efficiency of EAE protection. Administration of E(2) at pregnancy levels but not lower concentrations also enhanced the frequency of Treg cells. Additionally, E(2) treatment drastically reduced the production of interleukin-17 (IL-17) in the periphery of immunized mice. However, E(2) treatment did not protect against EAE or suppress IL-17 production in PD-1 gene-deficient mice. Finally, E(2) failed to prevent Treg-deficient mice from developing spontaneous EAE. Taken together, our results suggest that E(2)-induced protection against EAE is mediated by upregulation of PD-1 expression within the Treg-cell compartment. PMID- 19302145 TI - Secreted TARSH regulates olfactory mitral cell dendritic complexity. AB - Olfactory sensory neurons synapse with mitral cells to form stereotyped connections in the olfactory bulb (OB). Mitral cell apical dendrites receive input from olfactory sensory neurons expressing the same odorant receptor. During development, this restricted dendritic targeting of mitral cells is achieved through eliminating elaborated dendritic trees to a single apical dendrite. Through a genome-wide microarray screen, we identified TARSH (Target of NESH SH3) as a transiently expressed molecule in mitral cells during the dendritic refinement period. TARSH expression is restricted to pyramidal neurons along the main olfactory pathway, including the anterior olfactory nucleus and piriform cortex. The dynamic TARSH expression is not altered when odor-evoked activity is blocked by naris closure or in AC3 knockout mice. We also demonstrate that TARSH is a secreted protein. In dissociated OB cultures, secreted TARSH promotes the reduction of mitral cell dendritic complexity and restricts dendritic branching and outgrowth of interneurons. Dendritic morphological changes were also observed in mitral cells overexpressing TARSH themselves. We propose that TARSH is part of the genetic program that regulates mitral cell dendritic refinement. PMID- 19302146 TI - A putative role for cell cycle-related proteins in microtubule-based neuroplasticity. AB - Cyclins and cyclin-dependent kinases (Cdks) are the main components that control the orderly progression through cell cycle. In the mature nervous system, terminally differentiated neurons are permanently withdrawn from cell cycle, as mitotic quiescence is essential for the functional stability of the complexly wired neuronal system. Recently, we characterized the expression and colocalization of cyclins and Cdks in terminally differentiated pyramidal neurons. The functional impact of the expression of cell cycle-related proteins in differentiated neurons, however, has not been elucidated yet. In the present study, we show by immunoelectron microscopy and immunobiochemical methods an association of cyclins and Cdks with the microtubule network. Cyclins D, E, A and B as well as Cdks 1, 2 and 4 were also found to be associated with the microtubule-associated protein tau. Cyclin/Cdk complexes, in addition, exhibit kinase activity towards tau. In vitro, downregulation of cyclins and Cdks by a siRNA approach and by pharmacological inhibition promotes neurite extension. Taken together, these results indicate that the expression of cell cycle-related proteins in terminal differentiated neurons is associated with physiological functions beyond cell cycle control that might be involved in microtubule-based mechanisms of neuroplasticity. PMID- 19302147 TI - Glutamate induces directed chemotaxis of microglia. AB - Microglia in the brain possess dynamic processes that continually sample the surrounding parenchyma and respond to local insults by rapidly converging on the site of an injury. One of the chemotaxic agents responsible for this response is ATP. Here we show that the transmitter glutamate is another such chemotaxic agent. Microglia exposed to glutamate increase their cell membrane ruffling and migrate to a source of glutamate in cell culture and in spinal cord slices. This chemotaxis is meditated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and metabotropic glutamate receptors on the microglia. Chemotaxis is dependent on redistribution of actin filaments in the cells and on tubulin following receptor activation. Thus glutamate, which is released at synapses as well as from damaged cells, can mediate rapid chemotaxic responses from microglial cells. PMID- 19302148 TI - Neuroprotective activities of activated protein C mutant with reduced anticoagulant activity. AB - The anticoagulant activated protein C (APC) protects neurons and endothelium via protease activated receptor (PAR)1, PAR3 and endothelial protein C receptor. APC is neuroprotective in stroke models. Bleeding complications may limit the pharmacologic utility of APC. Here, we compared the 3K3A-APC mutant with 80% reduced anticoagulant activity and wild-type (wt)-APC. Murine 3K3A-APC compared with wt-APC protected mouse cortical neurons from N-methyl-D-aspartate-induced apoptosis with twofold greater efficacy and more potently reduced N-methyl-D aspartate excitotoxic lesions in vivo. Human 3K3A-APC protected human brain endothelial cells (BECs) from oxygen/glucose deprivation with 1.7-fold greater efficacy than wt-APC. 3K3A-APC neuronal protection required PAR1 and PAR3, as shown by using PAR-specific blocking antibodies and PAR1- and PAR3-deficient cells and mice. BEC protection required endothelial protein C receptor and PAR1. In neurons and BECs, 3K3A-APC blocked caspase-9 and -3 activation and induction of p53, and decreased the Bax/Bcl-2 pro-apoptotic ratio. After distal middle cerebral artery occlusion (dMCAO) in mice, murine 3K3A-APC compared with vehicle given 4:00 h after dMCAO improved the functional outcome and reduced the infarction volume by 50% within 3 days. 3K3A-APC compared with wt-APC multi dosing therapy at 12:00 h, 1, 3, 5 and 7 days after dMCAO significantly improved functional recovery and reduced the infarction volume by 75% and 38%, respectively, within 7 days. The wt-APC, but not 3K3A-APC, significantly increased the risk of intracerebral bleeding as indicated by a 50% increase in hemoglobin levels in the ischemic hemisphere. Thus, 3K3A-APC offers a new approach for safer and more efficacious treatments of neurodegenerative disorders and stroke with APC. PMID- 19302149 TI - Partial compensation for N-type Ca(2+) channel loss by P/Q-type Ca(2+) channels underlines the differential release properties supported by these channels at cerebrocortical nerve terminals. AB - N-type and P/Q-type Ca(2+) channels support glutamate release at central synapses. To determine whether the glutamate release mediated by these channels exhibits distinct properties, we have isolated each release component in cerebrocortical nerve terminals from wild-type mice by specifically blocking N type Ca(2+) channels with omega-conotoxin-GVIA and P/Q-type Ca(2+) channels with omega-agatoxin-IVA. In addition, we have determined the release properties at terminals from mice lacking the alpha(1B) subunit of N-type channels (Ca(v) 2.2) to test the possibility that P/Q-type channels can compensate for the loss of N type Ca(2+) channels. We recently demonstrated that, while evoked glutamate release depends on P/Q- and N-type channels in wild-type nerve terminals, only P/Q-type channels participate in these knockout mice. Moreover, in nerve terminals expressing solely P/Q-type channels, metabotropic glutamate receptor 7 (mGluR7) fails to inhibit the evoked Ca(2+) influx and glutamate release. Here, we show that the failure of mGluR7 to modulate evoked glutamate release is not due to a lack of receptors, as nerve terminals from mice lacking N-type Ca(2+) channels express mGluR7. Indeed, we show that other receptor responses, such as the inhibition of forskolin-induced release, are preserved in these knockout mice. N-type channels are more loosely coupled to release than P/Q-type channels in nerve terminals from wild-type mice, as reflected by the tighter coupling of release in knockout nerve terminals. We conclude that the glutamate release supported by N- and P/Q-type channels exhibits distinct properties, and that P/Q type channels cannot fully compensate for the loss of N-type channels. PMID- 19302151 TI - Rapid plasticity at inhibitory and excitatory synapses in the hippocampus induced by ictal epileptiform discharges. AB - Epileptic seizures can induce pathological processes of plasticity in the brain that tend to promote the generation of further seizures. However, the immediate impact of epileptic seizures on cellular excitability remains poorly understood. In order to unravel such early mechanisms of epilepsy-induced plasticity, we studied synaptic transmission before and shortly after three ictal discharges induced by transient elevation of extracellular K(+) in mouse hippocampal slices. Discharges were initiated in the CA3 region and propagated via the Schaffer collaterals into CA1 where they were associated with sustained membrane depolarization and bursts of action potentials in CA1 pyramidal cells. Subsequently, discharges were followed by long-term potentiation (LTP) of Schaffer collateral-evoked field excitatory post-synaptic potentials (EPSPs) in the CA1. The ability to generate epileptiform activity in response to repetitive stimulation was enhanced during LTP. Changes in both inhibitory and excitatory synaptic transmission contributed to LTP in CA1 pyramidal cells. Discharges reduced gamma-aminobutyric acid-A receptor-mediated hyperpolarizing inhibitory post-synaptic potentials by shifting their reversal potentials in a positive direction. At the same time, the amplitudes of Schaffer collateral-evoked RS alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor-mediated EPSPs and action potential-independent miniature EPSPs were enhanced. However, N-methyl d-aspartate receptor-mediated EPSPs remained unchanged. Paired-pulse stimulation revealed a reduced probability of glutamate release. Together, these changes in synaptic transmission produce a sustained increase in hippocampal excitability. We conclude that a few seizure-like ictal episodes are sufficient to cause fast and lasting changes in the excitation/inhibition balance in hippocampal networks, and therefore may contribute to early phases of progressive epileptogenesis. PMID- 19302150 TI - Induction and expression of GluA1 (GluR-A)-independent LTP in the hippocampus. AB - Long-term potentiation (LTP) at hippocampal CA3-CA1 synapses is thought to be mediated, at least in part, by an increase in the postsynaptic surface expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid (AMPA) receptors induced by N-methyl-d-aspartate (NMDA) receptor activation. While this process was originally attributed to the regulated synaptic insertion of GluA1 (GluR-A) subunit-containing AMPA receptors, recent evidence suggests that regulated synaptic trafficking of GluA2 subunits might also contribute to one or several phases of potentiation. However, it has so far been difficult to separate these two mechanisms experimentally. Here we used genetically modified mice lacking the GluA1 subunit (Gria1(-/-) mice) to investigate GluA1-independent mechanisms of LTP at CA3-CA1 synapses in transverse hippocampal slices. An extracellular, paired theta-burst stimulation paradigm induced a robust GluA1-independent form of LTP lacking the early, rapidly decaying component characteristic of LTP in wild-type mice. This GluA1-independent form of LTP was attenuated by inhibitors of neuronal nitric oxide synthase and protein kinase C (PKC), two enzymes known to regulate GluA2 surface expression. Furthermore, the induction of GluA1 independent potentiation required the activation of GluN2B (NR2B) subunit containing NMDA receptors. Our findings support and extend the evidence that LTP at hippocampal CA3-CA1 synapses comprises a rapidly decaying, GluA1-dependent component and a more sustained, GluA1-independent component, induced and expressed via a separate mechanism involving GluN2B-containing NMDA receptors, neuronal nitric oxide synthase and PKC. PMID- 19302152 TI - Light stimulation evokes two different calcium responses in Muller glial cells of the guinea pig retina. AB - Intracellular calcium responses are a characteristic of glial activation upon neuronal activity. In acutely isolated preparations of the guinea pig retina, Muller glial cells displayed cytosolic calcium rises in response to repetitive light stimulation. The calcium rises consisted of two components, a slowly developing immediate response that occurred simultaneously over the whole length of all Muller cell fibers and a delayed fast response that originated in the ganglion cell layer and spread as a wave through the bodies of some Muller cells toward the outer processes in the photoreceptor layer. The slow calcium response was evoked by photoreceptor-to-glia signaling, resulting in a glutamate transporter- and zinc-mediated alteration in the membrane potential and an influx of calcium from the extracellular space. The fast calcium response was evoked by a release of calcium from intracellular stores, probably after activation of purinergic receptors. The data suggest that light stimulation of the retina causes glial activation by alterations in both the membrane potential and receptor-mediated mechanisms. The former may be implicated in glial support of the neuronal signal transfer from photoreceptors to ganglion cells (glial forward signaling), whereas the latter may constitute a glial feedback signaling from ganglion cells to photoreceptors. PMID- 19302153 TI - Inhibition of thalamic excitability by 4,5,6,7-tetrahydroisoxazolo[4,5-c]pyridine 3-ol: a selective role for delta-GABA(A) receptors. AB - The sedative and hypnotic agent 4,5,6,7-tetrahydroisoxazolo[4,5-c]pyridine-3-ol (THIP) is a GABA(A) receptor (GABA(A)R) agonist that preferentially activates delta-subunit-containing GABA(A)Rs (delta-GABA(A)Rs). To clarify the role of delta-GABA(A)Rs in mediating the sedative actions of THIP, we utilized mice lacking the alpha(1)- or delta-subunit in a combined electrophysiological and behavioural analysis. Whole-cell patch-clamp recordings were obtained from ventrobasal thalamic nucleus (VB) neurones at a holding potential of -60 mV. Application of bicuculline to wild-type (WT) VB neurones revealed a GABA(A)R mediated tonic current of 92 +/- 19 pA, which was greatly reduced (13 +/- 5 pA) for VB neurones of delta(0/0) mice. Deletion of the delta- but not the alpha(1) subunit dramatically reduced the THIP (1 mum)-induced inward current in these neurones (WT, -309 +/- 23 pA; delta(0/0), -18 +/- 3 pA; alpha(1) (0/0), -377 +/- 45 pA). Furthermore, THIP selectively decreased the excitability of WT and alpha(1) (0/0) but not delta(0/0) VB neurones. THIP did not affect the properties of miniature inhibitory post-synaptic currents in any of the genotypes. No differences in rotarod performance and locomotor activity were observed across the three genotypes. In WT mice, performance of these behaviours was impaired by THIP in a dose-dependent manner. The effect of THIP on rotarod performance was blunted for delta(0/0) but not alpha(1) (0/0) mice. We previously reported that deletion of the alpha(1)-subunit abolished synaptic GABA(A) responses of VB neurones. Therefore, collectively, these findings suggest that extrasynaptic delta-GABA(A)Rs vs. synaptic alpha(1)-subunit-containing GABA(A)Rs of thalamocortical neurones represent an important molecular target underpinning the sedative actions of THIP. PMID- 19302154 TI - Tolerance to non-opioid analgesics in PAG involves unresponsiveness of medullary pain-modulating neurons in male rats. AB - Opiate analgesia can be hampered by a reduction in pharmacological effectiveness (tolerance), and this crucially depends on the periaqueductal gray matter (PAG). Non-opioids like metamizol (dipyrone) or aspirin also induce PAG-dependent analgesia and tolerance, but the neuronal bases of this tolerance are unknown. Metamizol is a pyrazolon derivative and cyclooxygenase inhibitor with widespread use as an analgesic in Europe and Latin America. Metamizol was microinjected into the PAG of awake male rats, and antinociception was assessed by the tail flick (TF) and hot plate (HP) tests. Microinjection twice daily for 2.5 days caused tolerance to metamizol. The rats were then anesthetized and recordings from pain facilitating on-cells and pain-inhibiting off-cells of the rostral ventromedial medulla (RVM) were performed. PAG microinjection of morphine or metamizol depresses on-cells, activates off-cells and thus inhibits nociception, including TF and HP. In metamizol-tolerant rats, however, PAG microinjection of metamizol failed to affect on- or off-cells, and this is interpreted as the reason for tolerance. In metamizol-tolerant rats morphine microinjection into PAG also failed to affect RVM neurons or nociception (cross-tolerance). In naive, non tolerant rats the antinociceptive effect of PAG-microinjected metamizol or morphine was blocked when CTOP, a mu-opioid antagonist, was previously microinjected into the same PAG site. These results emphasize a close relationship between opioid and non-opioid analgesic mechanisms in the PAG and show that, like morphine, tolerance to metamizol involves a failure of on- and off-cells to, respectively, disfacilitate and inhibit nociception. Cross tolerance between non-opioid and opioid analgesics should be important in the clinical setting. PMID- 19302155 TI - Examination of intravenous and intra-CSF protein delivery for treatment of neurological disease. AB - Mucopolysaccharidosis type IIIA is a neurodegenerative lysosomal storage disorder characterized by progressive loss of learned skills, sleep disturbance and behavioural problems. Absent or greatly reduced activity of sulphamidase, a lysosomal protein, results in intracellular accumulation of heparan sulphate. Subsequent neuroinflammation and neurodegeneration typify this and many other lysosomal storage disorders. We propose that intra-cerebrospinal fluid protein delivery represents a potential therapeutic avenue for treatment of this and other neurodegenerative conditions; however, technical restraints restrict examination of its use prior to adulthood in mice. We have used a naturally occurring Mucopolysaccharidosis type IIIA mouse model to determine the effectiveness of combining intravenous protein replacement (1 mg/kg) from birth to 6 weeks of age with intra-cerebrospinal fluid sulphamidase delivery (100 microg, fortnightly from 6 weeks) on behaviour, the level of heparan sulphate oligosaccharide storage and other neuropathology. Mice receiving combination treatment exhibited similar clinical improvement and reduction in heparan sulphate storage to those only receiving intra-cerebrospinal fluid enzyme. Reductions in micro- and astrogliosis and delayed development of ubiquitin positive lesions were seen in both groups. A third group of intravenous-only treated mice did not exhibit clinical or neuropathological improvements. Intra cerebrospinal fluid injection of sulphamidase effectively, but dose-dependently, treats neurological pathology in Mucopolysaccharidosis type IIIA, even when treatment begins in mice with established disease. PMID- 19302156 TI - 17beta-estradiol and enriched environment accelerate cognitive recovery after focal brain ischemia. AB - Cognitive impairments, including spatial memory and learning deficiencies, are common after ischemic stroke. Estrogen substitution improves cognitive functions in post-menopausal women and ovariectomized rodents, partially through induction of neuroplasticity in the hippocampal formation. Post-ischemic housing of male rats in an enriched environment (EE) improves functional outcome, without changing infarct volume. We hypothesized that 17beta-estradiol combined with an EE would accelerate cognitive recovery after focal brain ischemia in ovariectomized rats and that recovery would be related to altered expression of nerve growth factor-induced gene (NGFI)-A in the hippocampus. 17beta-estradiol or placebo pellets were implanted 6 h after transient middle cerebral artery occlusion. Two days later, rats were placed in an EE or a deprived environment (DE) for 6 weeks. At 5 weeks after middle cerebral artery occlusion, 17beta estradiol-treated rats housed in an EE showed improvements in cognitive function (i.e. shorter latency and path in the Morris water maze task) compared with placebo-treated animals housed in an EE. Furthermore, beneficial effects on latency and path were observed when comparing EE-housed vs. DE-housed 17beta estradiol-treated rats. When comparing 17beta-estradiol-treated EE-housed rats vs. placebo-treated DE-housed rats, pronounced effects on latency and path were observed. Infarct volumes did not differ between groups. 17beta-estradiol-treated EE-housed rats had significantly higher NGFI-A mRNA expression bilaterally in the cornu ammonis 1 region and in the ipsilateral dentate gyrus of the hippocampus, compared with placebo-treated EE-housed rats. In conclusion, 17beta-estradiol treatment combined with an EE improved recovery of cognitive function after experimental brain ischemia, putatively through the upregulation of NGFI-A in hippocampal subregions. PMID- 19302157 TI - Recovery of motor and cognitive function after cerebellar lesions in a songbird: role of estrogens. AB - In addition to its key role in complex motor function, the cerebellum is increasingly recognized to have a role in cognition. Songbirds are particularly good models for the investigation of motor and cognitive processes but little is known about the role of the songbird cerebellum in these processes. To explore cerebellar function in a songbird, we lesioned the cerebellum of adult female zebra finches and examined the effects on a spatial working memory task and on motor function during this task. There is evidence for steroid synthesis in the songbird brain and neurosteroids may have an impact on some forms of neural plasticity in adult songbirds. We therefore hypothesized that neurosteroids would affect motor and cognitive function after a cerebellar injury. We found that cerebellar lesions produced deficits in motor and cognitive aspects of a spatial task. In line with our prediction, birds in which estrogen synthesis was blocked had impaired performance in our spatial task compared with those that had estrogen synthesis blocked but estrogen replaced. There was no clear effect of estrogen replacement on motor function. We also found that lesions induced expression of the estrogen synthetic enzyme aromatase in reactive astrocytes and Bergmann glia around a cerebellar lesion. These data suggest that the cerebellum of songbirds mediates both motor and cognitive function and that estrogens may improve the recovery of cognitive aspects of cerebellar function after injury. PMID- 19302158 TI - GABA(A) receptors mediate the opposing roles of dopamine and the tegmental pedunculopontine nucleus in the motivational effects of ethanol. AB - Recent work has demonstrated that changes in ventral tegmental area (VTA) GABA(A) receptor ion conductance properties are responsible for switching morphine's positive reinforcing properties from a dopamine-independent to a dopamine dependent pathway when an animal transitions from a non-deprived (minimal drug exposure) to a dependent (chronic drug exposure) and withdrawn state. Here we show that a double dissociation of ethanol's positive reinforcing properties is exactly opposite to that seen with morphine. In C57BL/6 mice, ethanol-conditioned place preferences were blocked in dopamine D2 receptor knockout non-deprived mice, but not by a lesion of the tegmental pedunculopontine nucleus (TPP). On the other hand, TPP lesions, but not a D2 receptor mutation, blocked ethanol conditioned place preferences in ethanol-dependent and withdrawn mice. The opposite effects of ethanol and opiates can be explained by their proposed actions through a common VTA GABA(A) receptor switching mechanism. PMID- 19302159 TI - Multisensory interactions within and outside the focus of visual spatial attention (Commentary on Fairhall & Macaluso). PMID- 19302160 TI - Spatial attention can modulate audiovisual integration at multiple cortical and subcortical sites. AB - The role of attention in multisensory integration (MI) is presently uncertain, with some studies supporting an automatic, pre-attentive process and others suggesting possible modulation through selective attention. The goal of this functional magnetic resonance imaging study was to investigate the role of spatial attention on the processing of congruent audiovisual speech stimuli (here indexing MI). Subjects were presented with two simultaneous visual streams (speaking lips in the left and right visual hemifields) plus a single central audio stream (spoken words). In the selective attention conditions, the auditory stream was congruent with one of the two visual streams. Subjects attended to either the congruent or the incongruent visual stream, allowing the comparison of brain activity for attended vs. unattended MI while the amount of multisensory information in the environment and the overall attentional requirements were held constant. Meridian mapping and a lateralized 'speaking-lips' localizer were used to identify early visual areas and to localize regions responding to contralateral visual stimulations. Results showed that attention to the congruent audiovisual stimulus resulted in increased activation in the superior temporal sulcus, striate and extrastriate retinotopic visual cortex, and superior colliculus. These findings demonstrate that audiovisual integration and spatial attention jointly interact to influence activity in an extensive network of brain areas, including associative regions, early sensory-specific visual cortex and subcortical structures that together contribute to the perception of a fused audiovisual percept. PMID- 19302161 TI - Contrast independence of cardinal preference: stable oblique effect in orientation maps of ferret visual cortex. AB - The oblique effect was first described as enhanced detection and discrimination of cardinal orientations compared with oblique orientations. Such biases in visual processing are believed to originate from a functional adaptation to environmental statistics dominated by cardinal contours. At the neuronal level, the oblique orientation effect corresponds to the numerical overrepresentation and narrower tuning bandwidths of cortical neurons representing the cardinal axes. The anisotropic distribution of orientation preferences over large cortical regions was revealed with optical imaging, providing further evidence for the cortical oblique effect in several mammalian species. Our present study explores whether the dominant representation of cardinal contours persists at different stimulus contrasts. Performing intrinsic optical imaging in the ferret visual cortex and presenting drifting gratings at various orientations and contrasts (100%, 30% and 10%), we found that the overrepresentation of vertical and horizontal contours was invariant across stimulus contrasts. In addition, the responses to cardinal orientations were also more robust and evoked larger modulation depths than responses to oblique orientations. We conclude that orientation maps remain constant across the full range of contrast levels down to detection thresholds. Thus, a stable layout of the functional architecture dedicated to processing oriented edges seems to reflect a fundamental coding strategy of the early visual cortex. PMID- 19302162 TI - Interhemispheric balance of overt attention: a theta burst stimulation study. AB - Interhemispheric imbalance is discussed as a pathophysiological mechanism in visuospatial neglect. It is suggested that after a lesion of the right hemisphere the mutual transcallosal inhibition is impaired, resulting in an increased activity of the left hemisphere. We investigated the interhemispheric balance of attention in healthy subjects by using a free visual exploration task and by interfering with the neural activity of the posterior parietal cortex (PPC) of either hemisphere using an inhibitory transcranial magnetic stimulation routine with theta burst stimulation (TBS). Subjects explored colour photographs of real life scenes presented on a computer screen under four conditions: (i) without TBS; (ii) after TBS over the right PPC; (iii) after TBS over the left PPC; and (iv) after TBS over the right PPC and, after the first half of the task, over the left PPC. Eye movements were measured, and distribution of mean cumulative fixation duration over screen halves was analyzed. TBS over the right PPC resulted in a significant rightward shift of mean cumulative fixation duration of approximately 30 min. The shift could be reversed when a subsequent train of TBS was applied over the left PPC. However, left PPC stimulation alone had no significant effect on visual exploration behaviour. The present study shows that the effect of TBS on the PPC depends on which hemisphere is stimulated and on the state of the contralateral homologue area. These findings are in accordance with the predictions of the interhemispheric rivalry model in neglect. PMID- 19302163 TI - Basal ganglia and frontal involvement in self-generated and externally-triggered finger movements in the dominant and non-dominant hand. AB - Although there are a number of functional neuroimaging studies that have investigated self-initiated and externally-triggered movements, data directly comparing right and left hands in this context are very scarce. The goal of this study was to further understand the role of the basal ganglia and prefrontal cortex in the realm of self-initiated and externally-triggered right and left hand movements. Young healthy right-handed adults performed random, follow and repeat conditions of a finger moving task with their right and left hands, while being scanned with functional magnetic resonance imaging. Significant activation of the dorsolateral prefrontal cortex was observed when comparing the self initiated movements with the repeated control and externally-triggered movements when using either hand in agreement with its role in monitoring. The caudate nucleus activation was found during self-initiated conditions compared with the control condition when either hand was used, showing that it is particularly involved when a new movement needs to be planned. Significant putamen activation was observed in all within-hand contrasts except for the externally-triggered vs. control condition when using the left hand. Furthermore, greater putaminal activation was found for the left vs. the right hand during the control condition, but for the right vs. the left hand subtraction for the self-initiated condition. Our results show that the putamen is particularly involved in the execution of non-routine movements, especially if those are self-initiated. Furthermore, we propose that, for right-handed people performing fine movements, as far as putamen involvement is concerned, the lack of proficiency of the non dominant hand may prevail over other task demands. PMID- 19302164 TI - The neuroanatomy of grapheme-color synesthesia. AB - Grapheme-color synesthetes perceive particular colors when seeing a letter, word or number (grapheme). Functional neuroimaging studies have provided some evidence in favor of a neural basis for this type of synesthesia. Most of these studies have reported extra activations in the fusiform gyrus, which is known to be involved in color, letter and word processing. The present study examined different neuroanatomical features (i.e. cortical thickness, cortical volume and cortical surface area) in a sample of 48 subjects (24 grapheme-color synesthetes and 24 control subjects), and revealed increased cortical thickness, volume and surface area in the right and left fusiform gyrus and in adjacent regions, such as the lingual gyrus and the calcarine cortex, in grapheme-color synesthetes. In addition, we set out to analyze structural connectivity based on fractional anisotropy (FA) measurements in a subsample of 28 subjects (14 synesthetes and 14 control subjects). In contrast to the findings of a recent neuroanatomical study using modern diffusion tensor imaging measurement techniques, we did not detect any statistically significant difference in FA between synesthetes and non synesthetes in the fusiform gyri. Our study thus supports the hypothesis of local anatomical differences in cortical characteristics in the vicinity of the V4 complex. The observed altered brain anatomy in grapheme-color synesthetes might be the anatomical basis for this particular form of synesthesia but it is also possible that the detected effects are a consequence (rather than the primary cause) of the life-long experience of grapheme-color synesthesia. PMID- 19302165 TI - Functional connectivity during a social emotion task in adolescents and in adults. AB - In this fMRI study we investigated functional connectivity between components of the mentalising system during a social emotion task, using psychophysiological interaction (PPI) analysis. Ten adults (22-32 years) and 18 adolescents (11-18 years) were scanned while thinking about scenarios in which a social or a basic emotion would be experienced. Unlike basic emotions (such as disgust and fear), social emotions (such as embarrassment and guilt) require the representation of another's mental states. In both adults and adolescents, an anterior rostral region of medial prefrontal cortex (arMPFC) involved in mentalising showed greater connectivity with the posterior superior temporal sulcus (pSTS) bordering on the temporo-parietal junction (TPJ) and with anterior temporal cortex (ATC) during social than during basic emotion. This result provides novel evidence that components of the mentalising system interact functionally during a social emotion task. Furthermore, functional connectivity differed between adolescence and adulthood. The adolescent group showed stronger connectivity between arMPFC and pSTS/TPJ during social relative to basic emotion than did the adult group, suggestive of developmental changes in functional integration within the mentalising system. PMID- 19302166 TI - Oculomotor control in children with fetal alcohol spectrum disorders assessed using a mobile eye-tracking laboratory. AB - Prenatal exposure to alcohol can result in a spectrum of adverse developmental outcomes, collectively termed fetal alcohol spectrum disorders (FASDs). This study evaluated deficits in sensory, motor and cognitive processing in children with FASD that can be identified using eye movement testing. Our study group was composed of 89 children aged 8-15 years with a diagnosis within the FASD spectrum [i.e. fetal alcohol syndrome (FAS), partial fetal alcohol syndrome (pFAS), and alcohol-related neurodevelopmental disorder (ARND)], and 92 controls. Subjects looked either towards (prosaccade) or away from (antisaccade) a peripheral target that appeared on a computer monitor, and eye movements were recorded with a mobile, video-based eye tracker. We hypothesized that: (i) differences in the magnitude of deficits in eye movement control exist across the three diagnostic subgroups; and (ii) children with FASD display a developmental delay in oculomotor control. Children with FASD had increased saccadic reaction times (SRTs), increased intra-subject variability in SRTs, and increased direction errors in both the prosaccade and antisaccade tasks. Although development was associated with improvements across tasks, children with FASD failed to achieve age-matched control levels of performance at any of the ages tested. Moreover, children with ARND had faster SRTs and made fewer direction errors in the antisaccade task than children with pFAS or FAS, although all subgroups were different from controls. Our results demonstrate that eye tracking can be used as an objective measure of brain injury in FASD, revealing behavioral deficits in all three diagnostic subgroups independent of facial dysmorphology. PMID- 19302167 TI - Burning mouth syndrome: the role of contact hypersensitivity. AB - BACKGROUND: Burning mouth syndrome is a burning sensation or stinging disorder affecting the oral mucosa in the absence of any clinical signs or mucosal lesions. Some studies have suggested that burning mouth syndrome could be caused by the metals used in dental prostheses, as well as by acrylate monomers, additives and flavouring agents, although others have not found any aetiologic role for hypersensitivity to dental materials. OBJECTIVE: To evaluate the extent and severity of adverse reactions to dental materials in a group of patients with burning mouth syndrome, and investigate the possible role of contact allergy in its pathogenesis. MATERIALS AND METHODS: We prospectively studied 124 consecutive patients with burning mouth syndrome (108 males; mean age 57 years, range 41-83), all of whom underwent allergen patch testing between 2004 and 2007. RESULTS: Sixteen patients (13%) showed positive patch test reactions and were classified as having burning mouth syndrome type 3 or secondary burning mouth syndrome (Lamey's and Scala's classifications). CONCLUSION: Although we did not find any significant association between the patients and positive patch test reactions, it would be advisable to include hypersensitivity to dental components when evaluating patients experiencing intermittent oral burning without any clinical signs. PMID- 19302168 TI - Rootstock-mediated changes in xylem ionic and hormonal status are correlated with delayed leaf senescence, and increased leaf area and crop productivity in salinized tomato. AB - Tomato crop productivity under salinity can be improved by grafting cultivars onto salt-tolerant wild relatives, thus mediating the supply of root-derived ionic and hormonal factors that regulate leaf area and senescence. A tomato cultivar was grafted onto rootstocks from a population of recombinant inbred lines (RILs) derived from a Solanum lycopersicum x Solanum cheesmaniae cross and cultivated under moderate salinity (75 mM NaCl). Concentrations of Na(+), K(+) and several phytohormones [abscisic acid (ABA); the cytokinins (CKs) zeatin, Z; zeatin riboside, ZR; and the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC)] were analysed in leaf xylem sap in graft combinations of contrasting vigour. Scion leaf area correlated with photosystem II (PSII) efficiency (F(v)/F(m)) and determined fruit productivity. Xylem K(+) (but not Na(+)), K(+)/Na(+), the active CK Z, the ratio with its storage form Z/ZR and especially the ratio between CKs and ACC (Z/ACC and Z + ZR/ACC) were positively loaded into the first principal component (PC) determining both leaf growth and PSII efficiency. In contrast, the ratio ACC/ABA was negatively correlated with leaf biomass. Although the underlying physiological mechanisms by which rootstocks mediate leaf area or chlorophyll fluorescence (and thus influence tomato salt tolerance) seem complex, a putative potassium-CK interaction involved in regulating both processes merits further attention. PMID- 19302169 TI - Temperature-induced lipocalin is required for basal and acquired thermotolerance in Arabidopsis. AB - Plant temperature-induced lipocalins (TILs) have been shown to be responsive to heat stress (HS), but the nature of this response was unknown. In this study, a reverse genetic approach was taken to elucidate the role of Arabidopsis TIL1 (At5g58070) in thermotolerance. A T-DNA knock-out line of TIL1 (til1-1) showed severe defects in basal (BT) and acquired thermotolerance (AT), which could be complemented by introducing the wild-type gene. However, over-expression of TIL1 did not significantly enhance thermotolerance in transgenic plants. TIL1 is peripherally associated with plasma membrane. Transcriptomic analysis showed that the heat shock response in til1-1 seedlings was about the same as in the wild type plants except the expression of TIL1. The level of TIL1 did not affect the temperature threshold for heat shock protein induction. Ion leakage analysis revealed no significant difference in membrane stability between the wild-type and til1-1 seedlings. These results suggest that TIL1 is not involved in regulating membrane fluidity or stability. Nevertheless, the mutant plants were also more sensitive than the wild type to tert-butyl hydroperoxide, a reagent that induces lipid peroxidation. Taken together, these data indicate that TIL1 is an essential component for thermotolerance and probably functions by acting against lipid peroxidation induced by severe HS. PMID- 19302170 TI - The plasma membrane Na+/H+ antiporter SOS1 is essential for salt tolerance in tomato and affects the partitioning of Na+ between plant organs. AB - We have identified a plasma membrane Na(+)/H(+) antiporter gene from tomato (Solanum lycopersicum), SlSOS1, and used heterologous expression in yeast to confirm that SlSOS1 was the functional homolog of AtSOS1. Using post transcriptional gene silencing, we evaluated the role played by SlSOS1 in long distance Na(+) transport and salt tolerance of tomato. Tomato was used because of its anatomical structure, more complex than that of Arabidopsis, and its agricultural significance. Transgenic tomato plants with reduced expression of SlSOS1 exhibited reduced growth rate compared to wild-type (WT) plants in saline conditions. This sensitivity correlated with higher accumulation of Na(+) in leaves and roots, but lower contents in stems of silenced plants under salt stress. Differential distribution of Na(+) and lower net Na(+) flux were observed in the xylem sap in the suppressed plants. In addition, K(+) concentration was lower in roots of silenced plants than in WT. Our results demonstrate that SlSOS1 antiporter is not only essential in maintaining ion homeostasis under salinity, but also critical for the partitioning of Na(+) between plant organs. The ability of tomato plants to retain Na(+) in the stems, thus preventing Na(+) from reaching the photosynthetic tissues, is largely dependent on the function of SlSOS1. PMID- 19302171 TI - Ozone suppresses soil drying- and abscisic acid (ABA)-induced stomatal closure via an ethylene-dependent mechanism. AB - Elevated atmospheric ozone concentrations (70 ppb) reduced the sensitivity of stomatal closure to abscisic acid (ABA) in Leontodon hispidus after at least 24 h exposure (1) when detached leaves were fed ABA, and (2) when intact plants were sprayed or injected with ABA. They also reduced the sensitivity of stomatal closure to soil drying around the roots. Such effects could already be occurring under current northern hemisphere peak ambient ozone concentrations. Leaves detached from plants which had been exposed to elevated ozone concentrations generated higher concentrations of ethylene, although leaf tissue ABA concentrations were unaffected. When intact plants were pretreated with the ethylene receptor binding antagonist 1-methylcyclopropene, the stomatal response to both applied ABA and soil drying was fully restored in the presence of elevated ozone. Implications of ethylene's antagonism of the stomatal response to ABA under oxidative stress are discussed. We suggest that this may be one mechanism whereby elevated ozone induces visible injury in sensitive species. We emphasize that drought linked to climate change and tropospheric ozone pollution, are both escalating problems. Ozone will exacerbate the deleterious effects of drought on the many plant species including valuable crops that respond to this pollutant by emitting more ethylene. PMID- 19302172 TI - Local and systemic effects of two herbivores with different feeding mechanisms on primary metabolism of cotton leaves. AB - Caterpillars and spider mites are herbivores with different feeding mechanisms. Spider mites feed on the cell content via stylets, while caterpillars, as chewing herbivores, remove larger amounts of photosynthetically active tissue. We investigated local and systemic effects of short-term caterpillar and spider mite herbivory on cotton in terms of primary metabolism and growth processes. After short-term caterpillar feeding, leaf growth and water content were decreased in damaged leaves. The glutamate/glutamine ratio increased and other free amino acids were also affected. In contrast, mild spider mite infestation did not affect leaf growth or amino acid composition, but led to an increase in total nitrogen and sucrose concentrations. Both herbivores induced locally increased dark respiration, suggesting an increased mobilization of storage compounds potentially available for synthesis of defensive substances, but did not affect assimilation and transpiration. Systemically induced leaves were not significantly affected by the treatments performed in this study. The results show that cotton plants do not compensate the loss of photosynthetic tissue with higher photosynthetic efficiency of the remaining tissue. However, early plant responses to different herbivores leave their signature in primary metabolism, affecting leaf growth. Changes in amino acid concentrations, total nitrogen and sucrose content may affect subsequent herbivore performance. PMID- 19302173 TI - Fog interception by Sequoia sempervirens (D. Don) crowns decouples physiology from soil water deficit. AB - Although crown wetting events can increase plant water status, leaf wetting is thought to negatively affect plant carbon balance by depressing photosynthesis and growth. We investigated the influence of crown fog interception on the water and carbon relations of juvenile and mature Sequoia sempervirens trees. Field observations of mature trees indicated that fog interception increased leaf water potential above that of leaves sheltered from fog. Furthermore, observed increases in leaf water potential exceeded the maximum water potential predicted if soil water was the only available water source. Because field observations were limited to two mature trees, we conducted a greenhouse experiment to investigate how fog interception influences plant water status and photosynthesis. Pre-dawn and midday branchlet water potential, leaf gas exchange and chlorophyll fluorescence were measured on S. sempervirens saplings exposed to increasing soil water deficit, with and without overnight canopy fog interception. Sapling fog interception increased leaf water potential and photosynthesis above the control and soil water deficit treatments despite similar dark-acclimated leaf chlorophyll fluorescence. The field observations and greenhouse experiment show that fog interception represents an overlooked flux into the soil-plant-atmosphere continuum that temporarily, but significantly, decouples leaf-level water and carbon relations from soil water availability. PMID- 19302174 TI - Lung function at 10 yr is not impaired by early childhood lower respiratory tract infections. AB - The causal relationship between lower respiratory tract infections (LRIs) in early life and reduced lung function later in childhood is unsettled. Therefore, we assessed whether LRIs the first 2 yr of life influenced lung function development from birth to school age. In the prospective Oslo birth cohort, 'the Environment and Childhood Asthma (ECA) study' lung function was measured at birth in 802 infants by tidal flow volume loops and in 664 infants by passive respiratory mechanics and half yearly questionnaires, including LRI questions, were completed until 2 yr of age. The present study includes 607 children with information about LRIs the first 2 yr of life and successfully forced expiratory flow (FEF) volume measurements at the 10-yr follow-up assessment. At 10 yr of age, FEF at 50% of forced vital capacity (FEF(50)) (mean 95% confidence interval) was reduced in children with at least one bronchiolitis (85.0, 80.6-89.5, p = 0.020) or bronchitis (86.2, 82.6-89.8, p = 0.030) or > or =3 LRIs (83.4, 78.1 88.8, p = 0.017) when compared with no LRIs (90.6, 88.8-92.5) by 2 yr of life. The effects were significant in girls only when stratifying for gender. Among girls with later bronchiolitis compliance of the respiratory system (3.64, 3.17 4.10 vs. 4.18, 3.98-4.37, p = 0.031) and the ratio of time to peak tidal expiratory flow to total expiratory time (t(PTEF)/t(E)) measured at birth was significantly reduced (0. 26, 0.23-0.29 vs. 0.32, 0.30-0.33, p = 0.005) when compared with children with no LRIs. Change in lung function from birth (by t(PTEF)/t(E)) to 10 yr of age was not significantly associated with LRIs the first 2 yr of life, and LRIs by 2 yr of life were not significantly associated with lung function at 10 yr of age in regression analyses including lung function at birth and other possible predictors of lung function at 10 yr. In our study, LRIs during the first 2 yr of life did not impair lung function development from birth until 10 yr of age. PMID- 19302175 TI - Nasopharyngeal aspirate cytokine levels 1 yr after severe respiratory syncytial virus infection. AB - Respiratory syncytial virus (RSV) infection is an important cause of recurrent wheezing in infants. Nevertheless, the link between RSV infection and wheezing has yet to be elucidated at the molecular level. Here, we present a preliminary study on the evolution of the immune response in the respiratory tract at long term after RSV infection. Twenty-seven immune mediators were profiled in nasopharyngeal aspirates (NPAs) obtained from 20 children hospitalized due to a severe infection by RSV at discharge from hospital and again 1 yr later. The same mediators were profiled in parallel in NPAs from 12 healthy controls. In the year following discharge, 85% (17/20) of children of the RSV group suffered at least one episode of wheezing documented by the pediatrician. On the contrary, wheezing episodes were observed only in 25% (3/12) of children in the control group. While most of the mediators profiled returned to normal levels by 1 yr after discharge from hospital, RSV children showed a persistent nasal hyper-secretion of VEGF, G CSF, IL-10, IL-6, IFN-gamma, IL-7 and IL-13. In previous works VEGF, IL-10 and IFN-gamma have been put in relation with the pathogenesis of post-virus induced asthma. G-CSF, IL-6, IL-7 and IL-13 are increased in respiratory and plasma samples of asthmatic patients. Here, we evidence for the first time a persistent elevation of these mediators as late as 1 yr after severe RSV disease resolution, reinforcing their possible implication in the pathogenesis of wheezing. PMID- 19302176 TI - The regulatory function of umbilical cord blood CD4(+) CD25(+) T cells stimulated with anti-CD3/anti-CD28 and exogenous interleukin (IL)-2 or IL-15. AB - The abundance of CD4(+) CD25(+) regulatory T cells in umbilical cord blood (UCB) might contribute to the decreased severity of graft-vs.-host disease (GVHD) for UCB transplantation. This study aims to characterize the phenotypes and suppressive function of UCB CD4(+) CD25(+) T cells under the influence of anti CD3/anti-CD28 (CD3/CD28) and exogenous interleukin (IL)-2 or IL-15. Higher percentages of CD4(+) CD25(high) and FoxP3(+) cells were detected in UCB compared to their adult counterparts. IL-15 was as effective as IL-2 in enhancing the proliferation of CD3/CD28 stimulated UCB CD4(+) CD25(+) T cells. Phenotypically, IL-2/IL-15-stimulated UCB CD4(+) CD25(+) T cells expressed higher level of CTLA 4, GITR, membrane bound transforming growth factor-beta (mTGF-beta), and especially Foxp-3 than controls. IL-2/IL-15-stimulated UCB CD4(+) CD25(+) T cells also produced much higher IL-10 and TGF-beta than controls; while IL-2/IL-15 stimulated UCB CD4(+) CD25(-) T cells showed increased TGF-beta, but not IL-10 production. IL-2/IL-15-cultured UCB CD4(+) CD25(+) T cells showed comparable suppressor activity on allogeneic adult CD4(+) T-cell proliferation compared to controls, partly through a contact-dependent fashion. Taken together, IL-2/IL-15 stimulated UCB CD4(+) CD25(+) T cells show distinct regulatory T-cell phenotypic and functional features, and may be applied for the alleviation of GVHD severity following UCB transplantation. PMID- 19302177 TI - Effect of arch length on the functional well-being of dentate adults. AB - The aim of this study was to assess the influence of arch length and the number of occlusal units on the oral functions and general satisfaction perceived by dentate adults without dentures who had all their aesthetic units intact. We performed an epidemiological study at randomly chosen health centres on populations of adults without dentures. The subjects (n = 624) were classified as complete dental arch (CDA), interrupted dental arch (IDA) or shortened dental arches (SDA), depending on the length and continuity of the dental arches. We gathered clinical data and data on functional ability and oral satisfaction, plotting them on a scale of 0-10. The individuals with a shortened dental arch were found to have longer-lasting, more frequent and more severe functional limitations upon chewing, smiling and speaking than those with a complete or interrupted arch. The prevalance rate ratio (95% CI) of functional limitations in the SDA group was higher than in the aggregated CDA-IDA group, the values ranging between 1.56 (1.22-12.01) as regards chewing and 2.35 (1.45-3.85) in the case of smiling. However, in general all groups were satisfied with their oral status. PMID- 19302178 TI - Are true multihost fungi the exception or the rule? Dominant ectomycorrhizal fungi on Pinus sabiniana differ from those on co-occurring Quercus species. PMID- 19302179 TI - Eradication of hepatitis C virus by interferon improves whole-body insulin resistance and hyperinsulinaemia in patients with chronic hepatitis C. AB - BACKGROUND/AIMS: To investigate whether eradication of hepatitis C virus (HCV) by interferon (IFN) therapy influences systemic glucose metabolism. METHODS: Seventy two patients with chronic hepatitis C were enrolled in this study. Patients received IFN therapy and were classified into two groups: sustained responders (n=48) and nonsustained responders (n=24). We analysed systemic glucose metabolism in terms of the following indices: homeostasis model assessment for insulin resistance (HOMA-IR) and beta-cell function (HOMA-beta), insulinogenic index (II), composite insulin sensitivity index (ISI composite) and the area under the curve of plasma glucose (PG-AUC) and serum insulin (SI-AUC) in oral glucose tolerance tests. In 28 sustained responders and 16 nonsustained responders, serum levels of soluble tumour necrosis factor receptor 2 (sTNFR2) were measured. Indices were determined before and 6 months after therapy. RESULTS: In the sustained responders, HOMA-beta (P=0.0004) and SI-AUC (P=0.002) were significantly decreased and the ISI composite was increased (P=0.009), although there were no significant changes in HOMA-IR, II or PG-AUC. Serum sTNFR2 levels decreased significantly after therapy in sustained responders (P=0.001). In the nonsustained responders, there were no changes in any index. CONCLUSIONS: Eradication of HCV by IFN therapy could improve whole-body insulin resistance and insulin hypersecretion with reduced serum TNF-alpha levels. PMID- 19302180 TI - Impaired liver regeneration with humoral and genetic disturbances in urinary trypsin inhibitor-deficient mice. AB - BACKGROUND/AIMS: Urinary trypsin inhibitor (UTI) is an innate anti-inflammatory regulator. It can block the release of inflammatory factors, prevent the cascade reaction of cytokines and inhibit excessive activation of leukocytes. Liver regeneration (LR) is a dynamic molecular phenomenon without inflammation. Many cytokines, including tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), have been implicated in regulating LR. However, the role of UTI in LR is totally unknown. The aim of this study was to elucidate the role of UTI in LR using genetically UTI-deficient mice. METHODS: We performed 68% hepatectomy, comparing UTI (-/-) and UTI (+/+) mice. Recovery of liver weight was recorded and we calculated labelling indices after 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry. A DNA microarray was used to examine gene expression followed by real-time polymerase chain reaction. Serum IL-6, IL-10, monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1beta (MIP 1beta) were measured. RESULTS: LR in UTI (-/-) mice was delayed at 36 h after hepatectomy, at which time the DNA profile was different. One hundred and fourteen genes were upregulated and 100 genes were downregulated in UTI (-/-) mice at 36 h after hepatectomy among the 21, 977 mRNAs examined. Furthermore, serum IL-6, IL-10, MCP-1 and MIP-1beta levels at 36 h after hepatectomy in the UTI (-/-) mice were significantly higher than in the UTI (+/+) mice. CONCLUSION: UTI appears to important cytokine and chemokine regulation in normal liver regeneration. PMID- 19302181 TI - Effect of hyponatraemia on outcomes following orthotopic liver transplantation. AB - BACKGROUND: Hyponatraemia increases risk of adverse outcomes following orthotopic liver transplantation (OLT), but it is unclear whether improvement of pretransplant hyponatraemia ameliorates post-transplant complications. AIMS: To assess impact of pretransplant hyponatraemia on post-transplant outcomes. METHODS: We performed a retrospective analysis of 213 patients with cirrhosis who underwent liver transplantation. Patients with serum sodium 130 mEq/L at transplantation ('resolved hyponatraemia'; n=56) and to those without history of hyponatraemia before transplantation ('never hyponatraemic'; n=123). Primary endpoint was survival at 180 days post-OLT. Secondary outcomes included time until discharge alive, complications during hospitalization, length of time ventilated and length of post-transplant intensive care unit stay. RESULTS: There was no survival difference at 180 days post-OLT between groups. After transplantation, patients with either hyponatraemia at OLT or resolved hyponatraemia had longer time until discharge alive and had higher rates of delirium, acute renal failure, acute cellular rejection and infection than those who were never hyponatraemic. As compared with patients with hyponatraemia at OLT, those with resolved hyponatraemia were more likely to be discharged alive within 3 weeks, but other outcomes, including survival, did not differ significantly. CONCLUSIONS: We conclude that hyponatraemia at any time before liver transplantation is associated with adverse post-transplant outcome, even when hyponatraemia has resolved. PMID- 19302182 TI - Association of inflammatory and anti-inflammatory cytokines with insulin resistance in chronic hepatitis C. AB - BACKGROUND: The pathogenetic basis for the association between hepatitis C virus (HCV) infection and type-2 diabetes remains uncertain. It has been reported that insulin resistance (IR) plays an essential role. We investigated the association of inflammatory [tumour necrosis factor (TNF)-alpha, interleukin (IL)-6] and anti inflammatory cytokines (adiponectin and IL-10) with IR in chronic HCV infection. METHODS: Eighty-one consecutive non-diabetic chronic hepatitis C patients (37 men and 44 women, mean age of 51.9+/-12.2 years) and 40 age, sex and body mass index (BMI)-matched healthy individuals were collected. IR was evaluated by the homoeostasis model assessment (HOMA). Serum levels of cytokines were measured by enzyme-linked immunosorbent assay. RESULTS: Patients with chronic hepatitis C have a higher HOMA-IR, TNF-alpha, IL-6, adiponectin and IL-10, as compared with controls. By multiple linear regression analysis, moderate/severe steatosis grade, total cholesterol level and adiponectin was significantly associated with HOMA-IR, whereas, TNF-alpha, IL-6 and IL-10 was not. Male gender, BMI and HOMA-IR was inversely correlated with the serum adiponectin level. Serum adiponectin was positively correlated with TNF-alpha level, which was significantly associated with higher degree of hepatic necroinflammation. CONCLUSION: Our data suggest that chronic HCV infection is associated with increased IR, which is correlated inversely with the serum adiponectin level. The complex role of adiponectin in the pathogenesis of IR and hepatic necroinflammation in chronic HCV infection merit further investigation. PMID- 19302183 TI - Primary cultures of human hepatocytes isolated from hepatitis C virus-infected cirrhotic livers as a model to study hepatitis C infection. AB - BACKGROUND/AIM: Since the discovery of hepatitis C virus (HCV), researchers have encountered difficulties with in vitro models. The aim of this study was to determine whether HCV-infected human primary hepatocytes, isolated from cirrhotic livers at liver transplantation, can be used as a model to study HCV infection. METHODS: Hepatocytes were isolated with collagenase and cultured over a 20-day period on different matrices. Viral kinetics was monitored with/without treatment by real-time polymerase chain reaction. RESULTS: Cell yield and viability were higher with uninfected/non-cirrhotic livers (77.2+/-1.8%) in comparison with HCV infected cirrhotic livers (68.8+/-12%). HCV-infected hepatocytes behaved similar to non-infected cells and expressed albumin and cytochrome P4502E1. HCV-positive strand was identified in supernatants and cell lysates. HCV-negative strand was only found inside cells and correlated with viral RNA recovery in the medium. Improvement in the degree of hepatocyte differentiation was associated with better HCV recovery. Antiviral treatment with interferon-alpha, EX4 and cyclosporine A induced significant reductions in HCV RNA. CONCLUSION: Primary cultures of HCV-infected human hepatocytes from end-stage cirrhotic livers is feasible, represents an excellent model to study specific virus-host interactions and can be used to assess viral replication. PMID- 19302184 TI - Association between angiotensin II type 1 receptor polymorphisms and the occurrence of nonalcoholic fatty liver disease. AB - BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver injury in many countries. Genetic factors are important for the development of NAFLD, as well as environmental factors. Recently an angiotensin II type 1 receptor (AGTR1) has been recognized as important in the aetiology of fibrosis in the liver. OBJECTIVE: In this study we investigated the association between angiotensin II type 1 receptor gene polymorphism (ATGR1) and NAFLD. METHODS: One hundred and sixty-seven NAFLD patients [106 with nonalcoholic steatohepatitis (NASH) and 61 with simple steatosis] with a positive diagnosis by liver biopsy and 435 healthy control subjects were recruited in this study. RESULTS: We investigated 12 single nucleotide polymorphisms (SNPs) of the ATGR1 gene, among which rs3772622 showed the lowest P-value of allele frequency model (P=0.0000012) with an odds ratio (95% confidence interval) of 1.95 (1.49-2.55). Five SNPs (rs3772622, rs3772633, rs2276736, rs3772630 and rs3772627) were significantly associated with NAFLD, even when the most conservative Bonferroni's correction was applied. Linkage disequilibrium analysis revealed that SNP rs3772622 and another four SNPs (rs3772633, rs2276736, rs3772630 and rs3772627) were in the same block. We investigated the association between rs3772622 genotypes and the fibrosis index. The results of the analysis revealed an additive increase of the fibrosis index in the patients with the A allele of rs3772622. CONCLUSIONS: This is the first report to demonstrate the genetic variations in ATGR1 that may influence the risk of NAFLD and liver fibrosis in NAFLD. PMID- 19302185 TI - Significance of antibodies to soluble liver antigen/liver pancreas: a large French study. AB - BACKGROUND: Antibodies to soluble liver antigen (SLA)/liver pancreas (LP) are generally considered as highly specific diagnostic markers of type 1 auto-immune hepatitis (AIH-1), and are particularly useful in patients without conventional antibodies. However, the presence of anti-SLA/LP in type 2 auto-immune hepatitis (AIH-2), primary sclerosing cholangitis (PSC) and hepatitis C has recently been reported. The aim was thus to describe the characteristics of anti-SLA/LP positive patients in the largest series reported to date. METHODS: Sera were selected from the period between 1998 and 2005, based on the presence of antibodies to SLA/LP detected by two methods. The clinical status of patients was determined from their medical records. RESULTS: Eighty-one anti-SLA/LP-positive patients with available clinical data were included: 89% (72/81) had a diagnosis of AIH-1, including 10 (12%) associated with cholestatic diseases (primary biliary cirrhosis in seven cases and PSC in three cases). Six patients (7%) suffered from another liver disease: hepatitis C (n=3) and drug-induced hepatitis (n=3). No specific diagnosis was made in three patients. CONCLUSIONS: Antibodies to SLA/LP are of a major diagnostic value for AIH-1, including paediatric forms and overlap syndromes with cholestatic diseases, but are not found in association with anti-liver/kidney/microsome type 1 or antibodies to liver cytosol type 1. They are rarely present in other liver diseases such as hepatitis C and drug induced hepatitis. PMID- 19302186 TI - Signalling pathway mediated by CXCR7, an alternative chemokine receptor for stromal-cell derived factor-1alpha, in AtT20 mouse adrenocorticotrophic hormone secreting pituitary adenoma cells. AB - Stromal cell-derived factor (SDF)-1 and its receptor, CXCR4, have been identified in both neurones and glia of many brain areas. Previous studies have mainly focused on the role of SDF-1 and CXCR4 in modulating the hypothalamic-pituitary axis and their possible involvement in the development of pituitary adenomas. An alternative SDF-1 receptor, CXCR7, has recently been identified, but it has not been studied in the context of pituitary adenomas. The present study aimed to investigate the distribution and function of CXCR7 in pituitary adenomas. The expression of CXCR7, normalised to beta-actin, was assessed by tissue microarray analysis of 62 adenomas, including 23 growth hormone (GH)-producing adenomas, 22 nonfunctioning adenomas, seven prolactin (PRL)-producing adenomas, six adrenocorticotrophic hormone-producing adenomas and four thyroid-stimulating hormone-producing adenomas. In vitro functional studies used RNA interference (RNAi) and cDNA microarray analysis to evaluate the CXCR7 signalling pathway in AtT-20 mouse pituitary adenoma cells treated with recombinant mouse SDF-1alpha and transfected with RNAi against Cxcr7 or control RNAi. In tissue microarray analysis, prominent expression of CXCR7 was observed in GH-producing adenomas and PRL-producing adenomas, and in macroadenomas (P < 0.05). Intracellular signalling via CXCR7 up-regulated Bub1, Cdc29 and Ccnb1, and down-regulated Asns, Gpt, Pycr1, Cars and Dars. The present study demonstrates that the SDF-1alpha / CXCR7 signalling pathway regulates genes involved in cell cycle control, amino acid metabolism and ligase activity, which comprise targets that are distinct from those of CXCR4. PMID- 19302187 TI - Primary culture of the isolated terminal nerve-gonadotrophin-releasing hormone neurones derived from adult teleost (dwarf gourami, Colisa lalia) brain for the study of peptide release mechanisms. AB - Terminal nerve (TN)-gonadotrophin-releasing hormone (GnRH) neurones are suggested to release GnRH peptides from widely-branched neural processes and the somatodendritic regions, depending on their firing activities. The released GnRH may exert its neuromodulatory actions on GnRH receptors located on various target neurones. The electrophysiological and morphological characteristics of TN-GnRH neurones, which are shared with other peptidergic neurones of vertebrate brains, are thought to represent general features of neuromodulatory and / or neurosecretory neurones. To address questions concerning the ways in which the electrical activities of peptidergic (TN-GnRH) neuronal somata affect GnRH release from different neuronal compartments, we established a primary culture system of TN-GnRH neurones, which will facilitate simultaneous recordings of various physiological signals from different compartments of a single TN-GnRH neurone cultured in a flat plane. The whole brain of an adult freshwater teleost, the dwarf gourami, was dissected out. The TN-GnRH neurones were then isolated and plated on a coverslip in culture medium. The isolated TN-GnRH neurones could be cultured for up to 2 weeks. In culture, the neurones grew both axon- and dendrite like neurites, and these processes were phenotypically similar to those found in situ. Unlike the neurones in situ, the cultured neurones had somewhat depolarised resting membrane potentials and showed no spontaneous discharge, which, however, should not be considered to comprise unhealthy culture conditions. Instead, they showed subthreshold spontaneous membrane potential oscillations and could be induced to fire in phasic or tonic patterns. In addition, stimulus-induced exocytotic events could be demonstrated in the soma and neurites using a fluorescent dye, FM1-43. Thus, the present isolated culture of TN-GnRH neurones will open up a wide range of possibilities for studying cellular mechanism of exocytosis, generation of spontaneous firing activity, and neurite outgrowth in peptidergic neurones. PMID- 19302188 TI - A single episode of restraint stress regulates central corticotrophin- releasing hormone receptor expression and binding in specific areas of the mouse brain. AB - The importance of restraining stress-induced activation of the hypothalamic pituitary-adrenocortical (HPA) system within tolerable limits requires efficient mechanisms for feedback inhibition. Recently, central corticotrophin-releasing hormone (CRH) receptor type 1 (CRHR1) has been shown to mediate HPA system feedback inhibition. To date, most of the data regarding stress-associated expression changes of CRHR1 and CRHR2 mRNA and their ligand CRH have been generated in rats. Taken considerable species differences into consideration, and with the growing importance of transgenic mice, a systematic analysis of the time course of expression changes of CRH and its two receptors in the mouse brain is needed to provide more insight into the regulation of the HPA system, both under physiological and pathophysiological conditions in this species. We analysed in detail the time course of expression changes of CRH, CRHR1 and CRHR2 mRNA after of restraint stress in mice in stress-relevant brain regions (paraventricular nucleus, hippocampus, neocortex). We could show a rapid, strong and long-lasting decrease in cortical and hippocampal CRHR1 mRNA expression after stress, whereas CRHR2 mRNA increased in the same neuroanatomical areas. In situ hybridisation analyses could be further confirmed at the protein level by CRH receptor autoradiography with changes in CRH binding that persisted even 7 days after a single episode of restraint stress. Our observation that stress has opposing effects on CRHR1 and CRHR2 neuronal systems supports the idea that regulation of the relative contribution of the two CRH receptors to brain CRH pathways may be essential in coordinating physiological responses to stress. We further hypothesise that the sustained alteration of CRH receptor expression and binding after a single episode of stress could mediate the long-term effects of stress on neuroendocrine function and emotional regulation. PMID- 19302189 TI - The trophic effects of oestrogen on male rat anterior pituitary lactotrophs. AB - Rapid but often transient changes in mitotic and apoptotic activity are important components of the pituitary response to changes in the hormonal environment. For example, bilateral adrenalectomy and orchidectomy each result in a wave of increased mitosis lasting approximately 1 week, mediated by the same population of trophically active and, to a large extent, endocrinologically inactive cells. By contrast to these tonic inhibitors of pituitary trophic activity, reports of a progressive increase in lactotroph numbers during pregnancy suggest that oestrogen is a potent and persistent pituitary mitogen. By comparing the amplitude and duration of male rat anterior pituitary mitotic responses to oestrogen treatment, to adrenalectomy, and to a combination of the two, the present study aimed to further clarify the characteristics of the oestrogen induced trophic response, in particular whether lactotrophs are the predominant cell type involved. Adrenalectomy produced a wave of increased mitotic activity, which resolved within 7 days as expected, whereas oestrogen induced a significant increase in mitotic activity, which was sustained for the 14-day duration of the study. The trophic effects of combining adrenalectomy and oestrogen treatment were not additive in that the statistically insignificant upward trend in mitotic index during the first few days compared to oestrogen treatment alone was entirely abolished by oestrogen pre-treatment. The increase in mitotic activity in lactotrophs induced by oestrogen either with or without adrenalectomy did not result in an increase in the relative size of the prolactin-positive compared to prolactin-negative pituitary parenchymal cell numbers by the end of the study. Despite the marked increase in the lactotroph population that is reported during pregnancy, these data indicate that at least the early (i.e. within 2 weeks) mitotic response to pharmacological doses of oestrogen increases mitotic activity in the lactotroph subpopulation by only 5-8% relative to other cellular subpopulations. Unexpectedly, the mitotic response to oestrogen principally occurs in non-prolactin-containing cells and results in the recruitment, amongst other trophically responsive populations, of the entire subpopulation of prolactin-, adrenocorticotrophic hormone- and luteinising hormone-negative cells that respond mitotically to adrenalectomy. Oestrogen therefore has a previously unrecognised non-cell type-specific trophic effect in the pituitary that obscures the relative expansion of the lactotroph population by inducing concurrent increases in numbers of prolactin-negative cells, the nature of which at least in part remains to be determined. PMID- 19302190 TI - Thyroid hormone receptor beta mediates acute illness-induced alterations in central thyroid hormone metabolism. AB - Acute illness in mice profoundly affects thyroid hormone metabolism in the hypothalamus and pituitary gland. It remains unknown whether the thyroid hormone receptor (TR)-beta is involved in these changes. In the present study, we investigated central thyroid hormone metabolism during lipopolysaccharide (LPS) induced illness in TRbeta(-/-) mice compared to wild-type (WT) mice. We administered a sublethal dose of LPS or saline to TRbeta(-/-) and WT mice. TRbeta(-/-) mice displayed higher basal levels of serum triiodothyronine (T(3)) and thyroxine (T(4)) compared to WT, reflecting thyroid hormone resistance. In the periventricular area of the hypothalamus, we observed a marked decrease in thyrotrophin-releasing hormone (TRH) mRNA expression in TRbeta(-/-) and WT mice at t = 4 h, coinciding with the peak in plasma corticosterone. The decrease in TRH mRNA persisted in WT, but not in TRbeta(-/-) mice at t = 24 h. By contrast, the increase of type 2 deiodinase (D2) mRNA already present at 4 h after LPS remained significant at 24 h in TRbeta(-/-), but not in WT mice. LPS decreased pituitary thyroid-stimulating hormone beta mRNA expression in WT at 24 h but not in TRbeta(-/-) mice. The peak in pituitary D2 expression at t = 4 h in WT was absent in TRbeta(-/-) mice. The relative decrease in plasma T(3) and T(4) upon LPS treatment was similar in both strains, although, at t = 24 h, plasma T(3) tended to be restored in TRbeta(-/-) mice. Our results suggest that TRbeta is involved in suppression of the central component of the hypothalamic-pituitary thyroid axis in acute illness. PMID- 19302191 TI - Loss of hypothalamic response to leptin during pregnancy associated with development of melanocortin resistance. AB - Hypothalamic leptin resistance during pregnancy is an important adaptation that facilitates the state of positive energy balance required for fat deposition in preparation for lactation. Within the arcuate nucleus, pro-opiomelanocortin (POMC) neurones and neuropeptide Y (NPY)/agouti-related gene protein (AgRP) neurones are first-order leptin responsive neurones involved in the regulation of energy balance. The present study aimed to investigate whether the regulation of these neuropeptides is disrupted during pregnancy in association with the development of leptin resistance. As measured by quantitative in situ hybridisation, POMC and AgRP mRNA levels were not significantly different during pregnancy, whereas NPY mRNA levels increased such that, by day 21 of pregnancy, levels were significantly higher than in nonpregnant, animals. These data suggest that these neurones were not responding normally to the elevated leptin found during pregnancy. To further characterise the melanocortin system during pregnancy, double-label immunohistochemistry was used to quantify leptin-induced phosphorylation of signal transducer and activator of transcription 3 (pSTAT3) in POMC neurones, using alpha-melanocyte-stimulating hormone (MSH) as a marker. The percentage of alpha-MSH neurones containing leptin-induced pSTAT3 did not significantly differ from nonpregnant animals, indicating that there was no change in the number of POMC neurones that respond to leptin during pregnancy. Treatment with alpha-MSH significantly reduced food intake in nonpregnant rats, but not in pregnant rats, indicating resistance to the satiety actions of alpha MSH during pregnancy. The data suggest that multiple mechanisms contribute to leptin resistance during pregnancy. As well as a loss of responses in first-order leptin-responsive neurones in the arcuate nucleus, there is also a downstream disruption in the melanocortin system. PMID- 19302192 TI - 17beta-Oestradiol indirectly inhibits thyrotrophin-releasing hormone expression in the hypothalamic paraventricular nucleus of female rats and blunts thyroid axis response to cold exposure. AB - Energy expenditure and thermogenesis are regultated by thyroid and sex hormones. Several parameters of hypothalamic-pituitary-thyroid (HPT) axis function are modulated by 17beta-oestradiol (E(2)) but its effects on thyrotrophin-releasing hormone (TRH) mRNA levels remain unknown. We evaluated, by in situ hybridisation and Northern bloting, TRH expression in the paraventricular nucleus of the hypothalamus (PVN) of cycling rats, 2 weeks-ovariectomised (OVX) and OVX animals injected s.c. during 1-4 days with E(2) (5, 50, 100 or 200 MUg / kg) (OVX-E). Serum levels of E(2), thyroid-stimulating hormone (TSH), prolactin, corticosterone and triiodothyronine (T(3)) were quantified by radioimmunoassay. Increased serum E(2) levels were observed after 4 days injection of 50 MUg / kg E(2) (to 68.5 +/- 4.8 pg / ml) in OVX rats. PVN-TRH mRNA levels were slightly higher in OVX than in virgin females at dioestrous 1 or pro-oestrous, decreasing proportionally to increased serum E(2) levels. E(2) injections augmented serum T(3), prolactin, and corticosterone levels. Serum TSH levels augmented with 4 days 50 MUg / kg E(2), but not with the higher doses that enhanced serum T(3) levels. Exposure to cold for 1 h resulted in marked HPT axis activation in OVX rats, increasing the levels of TRH mRNA along the rostro-caudal PVN areas, as well as serum TSH, T(3), corticosterone and prolactin levels. By contrast, no significant changes in any of these parameters were observed in cold-exposed OVX E (50 MUg / kg E(2)) rats. Very few PVN-TRHergic neurones expressed the oestrogen receptor type-alpha, suggesting that the effects of E(2) on PVN-TRH expression are indirect, most probably as a result of its multiple modulatory effects on circulating hormones and their receptor sensitivity. The blunted response of OVX E rats to cold coincides with the effects of E(2) on the autonomic nervous system and increased cold tolerance. PMID- 19302193 TI - Age- and hormone-regulation of N-methyl-D-aspartate receptor subunit NR2b in the anteroventral periventricular nucleus of the female rat: implications for reproductive senescence. AB - Glutamate, acting through its N-methyl-D-aspartate (NMDA) and non-NMDA receptors in the hypothalamus, regulates reproductive neuroendocrine functions via direct and indirect actions upon gonadotrophin-releasing hormone (GnRH) neurones. Previous studies indicate that the NMDA receptor subunit NR2b undergoes changes in protein and gene expression in the hypothalamus in general, and on GnRH neurones in particular, during reproductive ageing. In the present study, we examined whether the NR2b-expressing cell population, both alone and in association with the NR1 subunit (i.e. the latter subunit is necessary for a functional NMDA receptor), is altered as a function of age and / or steroid hormone treatment. Studies focused on the anteroventral periventricular (AVPV) nucleus of the hypothalamus, a region critically involved in the control of reproduction. Young (3-5 months), middle-aged (9-12 months), and aged (approximately 22 months) female rats were ovariectomised and, 1 month later, they were treated sequentially with oestradiol plus progesterone, oestradiol plus vehicle, or vehicle plus vehicle, then perfused. Quantitative stereologic analysis of NR2b-immunoreactive cell numbers in the AVPV showed an age-associated decrease in the density of NR2b-immunoreactive cells, but no effect of hormone treatment. In a second study, immunofluorescent double labelling of NR2b and NR1 was analysed by confocal microscopy of fraction volume, a semi-quantitative measure of fluorescence intensity. No effect of ageing was detected for immunofluorescent NR1 or NR2b alone, whereas the NR2b fraction volume increased in the oestradiol plus vehicle group. With ageing, the fraction volume of the NR2b/NR1-colocalised subunits increased. Together with the stereology results, this suggests that, although fewer cells express the NR2b subunit in the ageing AVPV, a greater percentage of these subunits are co-expressed with NR1. Our results suggest that the subunit composition of NMDA receptors in the AVPV undergo both age- and hormonal-regulation, which may be related to previous observations of changes in functional responses of reproductive neuroendocrine systems to NMDA receptor modulators with ageing. PMID- 19302194 TI - Identification of a cis-acting element involved in the regulation of BACE1 mRNA alternative splicing. AB - beta-site APP cleaving enzyme 1 (BACE1) is the transmembrane aspartyl protease that catalyzes the first cleavage step during proteolysis of the beta-amyloid precursor protein, a process involved in the pathogenesis of Alzheimer disease. BACE1 pre-mRNA undergoes complex alternative splicing, and cis-acting elements important for its regulation have not been identified. We constructed and compared several BACE1 minigenes and found that BACE1 sequence from exon 3 through exon 5 was required for minigenes to undergo correct splicing. Minigene splicing was validated by showing specific splicing inhibition upon splice site mutation. Furthermore, we showed that mutation of the minigene at a predicted exonic splicing enhancer in exon 4 of BACE1 increased exon 4 skipping. Therefore, we have for the first time found evidence of a regulatory site involved in BACE1 alternative splicing, and these data indicate that minor sequence changes can dramatically alter BACE1 alternative splicing. PMID- 19302195 TI - Lack of CB1 receptor activity impairs serotonergic negative feedback. AB - Serotonergic and endocannabinoid systems are important substrates for the control of emotional behaviour and growing evidence show an involvement in the pathophysiology of mood disorders. In the present study, the absence of the activity of the CB(1) cannabinoid receptor impaired serotonergic negative feedback in mice. Thus, in vivo microdialysis experiments revealed increased basal 5-HT extracellular levels and attenuated fluoxetine-induced increase of 5 HT extracellular levels in the prefrontal cortex of CB(1) knockout compared with wild-type mice. These observations could be related to the significant reduction in the 5-HT transporter binding site density detected in frontal cortex and hippocampus of CB(1) knockout mice. The lack of CB(1) receptor also altered some 5-HT receptors related to the 5-HT feedback. Extracellular recordings in the dorsal raphe nucleus (DRN) revealed that the genetic and pharmacological blockade of CB(1) receptor induced a 5-HT(1A) autoreceptor functional desensitization. In situ hybridization studies showed a reduction in the expression of the 5-HT(2C) receptor within several brain areas related to the control of the emotional responses, such as the DRN, the nucleus accumbens and the paraventricular nucleus of the hypothalamus, whereas an over-expression was observed in the CA3 area of the ventral hippocampus. These results reveal that the lack of CB(1) receptor induces a facilitation of the activity of serotonergic neurons in the DRN by altering different components of the 5-HT feedback as well as an increase in 5-HT extracellular levels in the prefrontal cortex in mice. PMID- 19302196 TI - The Parkinson disease-associated protein kinase LRRK2 exhibits MAPKKK activity and phosphorylates MKK3/6 and MKK4/7, in vitro. AB - Autosomal dominant mutations in the human Leucine-Rich Repeat Kinase 2 (LRRK2) gene represent the most common monogenetic cause of Parkinson disease (PD) and increased kinase activity observed in pathogenic mutants of LRRK2 is most likely causative for PD-associated neurotoxicity. The sequence of the LRRK2 kinase domain shows similarity to MAP kinase kinase kinases. Furthermore, LRRK2 shares highest sequence homology with mixed linage kinases which act upstream of canonical MAPKK and are involved in cellular stress responses. Therefore, we addressed the question if LRRK2 exhibits MAPKKK activity by systematically testing MAPKKs as candidate substrates, in vitro. We demonstrate that LRRK2 variants phosphorylate mitogen-activated protein kinase kinases (MAPKK), including MKK3 -4, -6 and -7. MKKs act upstream of the MAPK p38 and JNK mediating oxidative cell stress, neurotoxicity and apoptosis. The disease-associated LRRK2 G2019S and I2020T mutations show an increased phosphotransferase activity towards MKKs correlating with the activity shown for its autophosphorylation. Our findings present evidence of a new class of molecular targets for mutant LRRK2 that link to neurotoxicity, cellular stress, cytoskeletal dynamics and vesicular transport. PMID- 19302197 TI - Axillary artery cannulation for cardiopulmonary bypass during surgery on the ascending aorta and arch. AB - BACKGROUND: Performing axillary artery cannulation, during cardiopulmonary bypass in patients with an atherosclerotic ascending aorta or acute dissection of the ascending aorta and arch, is of growing interest. Our aim is to present our experience, to describe the surgical technique, and to demonstrate the sufficient cerebral and total body perfusion through axillary artery cannulation. PATIENTS AND METHODS: Twenty-two patients (17 male, five female) underwent surgical treatment with the axillary technique. The log euro SCORE ranged from 6.77% to 70% (mean 28.28). Nine of these patients underwent elective procedure. Eight underwent aortic surgery for pathologies of the aorta and in one patient we performed combined aortic valve replacement and coronary artery bypass grafting. Thirteen patients underwent emergency operation because of acute dissection of the aorta. Twelve of these patients had a type A dissection (according to Stanford classification) and one patient had a type B aortic dissection. RESULTS: The majority of complications were associated with ruptured dissection of the thoracoabdominal aorta and acute dissection of ascending aorta. Despite preoperative disease states that placed our patients at high risk of stroke and visceral end-organ injury, no clinically demonstrable permanent postoperative deficits were observed. Our patients had no neurological dysfunction, stroke, or other complications. CONCLUSIONS: Antegrade cerebral perfusion is of paramount importance in cases of aortic atherosclerosis or aortic dissection. The axillary artery provides an excellent site for safe antegrade perfusion, which may play a role in preventing stroke. PMID- 19302198 TI - Harvesting of the radial artery for coronary artery bypass grafting: comparison of ultrasonic harmonic scalpel dissector with the conventional technique. AB - OBJECTIVES: Use of the radial artery for coronary artery bypass grafting is getting more popular. We started routine use of the ultrasonic dissecting scalpel in harvesting radial arteries aiming to minimize harvesting time, improve graft quality, and reduce wound complications. METHODS: Radial artery harvesting technique using harmonic scalpel (HS; 43 patients) was compared with the conventional technique (Hemostatic clips and scissors; 53 patients). To avoid spasm, the radial artery was not skeletonized and papaverine was used to irrigate radial artery routinely in all patients. RESULTS: Compared to the conventional technique, radial artery harvesting using the HS has a significantly shorter harvesting time (25 minutes vs. 50 minutes, p < 0.001) and required a significantly smaller number of hemostatic clips (3 vs. 40, p < 0.001). In situ free blood flow was significantly higher in HS group (80 mL/min vs. 40 mL/min, p < 0.001). There was no forearm wound infection in the HS group. There was no graft failure, reoperation for bleeding, or hand ischemia with the use of either technique. CONCLUSION: Harvesting the radial artery using the HS is less time consuming and decreased the use of hemostatic clips rather atraumatic with good quality graft. PMID- 19302199 TI - Pericardiectomy: a functional anatomical perspective for the choice of left anterolateral thoracotomy. AB - Although pericardiectomy remains an established method for pericardial resection, the choice of surgical approach is not definitive. Within South Africa, surgical referral for tuberculosis-induced chronic constrictive pericarditis has not declined. Anecdotal reports have indicated good operative results that appear to show an association with choice of surgical technique used. This study aimed to provide a functional anatomical perspective for performance and recovery of the heart during pericardiectomy based on anatomical dissection and surgical notes. En bloc specimens were harvested from 16 fresh cadavers and pericardial segments were measured in terms of percentage cover over surface area of the myocardium. Retrospective analysis of 116 surgical reports of pericardiectomy performed over a period of 20 years was conducted. Surgical notes were compared for median sternotomy and anterolateral left thoracotomy. Results from anatomical study indicated that although the anterior pericardium between the phrenic nerves constitutes about 58% of total selected pericardial area, total pericardium accessible over left ventricle from that approach was only 26%. When orientated in left anterolateral position, total accessible area of left ventricular pericardium was 37%. Standard deviations were found to be comparable. Means were significantly different, indicating that the left anterolateral approach allows wider access to the left ventricle. This paper provides a functional anatomical perspective for the choice of left anterolateral thoracotomy as a surgical approach for pericardiectomy. PMID- 19302200 TI - Use of mouthguard rates among university athletes during sport activities in Erzurum, Turkey. AB - The purpose of this study was to determine the attitudes of mouthguard users in Erzurum, Turkey. The participants' appreciation in the use of protective devices, such as mouthguards, during sport activities was also evaluated. In this study, a 10-item questionnaire was distributed to 50 coaches and a 10-item questionnaire was distributed to 768 university athletes of three different sport modalities (basketball, soccer and volleyball) in the Ataturk University Sport Competitions. The Sport Competitions are composed of 20 faculty, except the faculty of dentistry. The result of the coaches' questionnaires indicated that none of the athletes used mouthguards while participating in sports. Of the coaches, 64% had seen orofacial trauma in their athletes during sport activities and 76% believed that mouthguards prevented oral injuries. Of the coaches, 76% reported that university athletes should use mouthguards in sport activities. The result of the university athletes' questionnaires revealed that the mouthguard utilization rate was 0%. Of all players, 78.1% were males (age 17-29) and 21.9% were females (age 18-23). Of all players, 7.31% had suffered from one or more type of oral injury while not wearing mouthguards. The results indicate that in Turkey, the use of mouthguards is rare in sports. It should be a combined duty of dentists, sports physicians, and coaches to encourage the use of mouthguards during training and sport activities. Doctors and dentists need to recommend a more intensive education of students in sports medicine and sports dentistry. PMID- 19302201 TI - A retrospective study on the prognosis of teeth with root fracture in patients during the maintenance phase of periodontal therapy. AB - Teeth with root fracture generally have a poor prognosis requiring extraction; however, some patients do not wish to have these teeth extracted. Dentists do not have enough information regarding the prognosis of teeth with root fracture. This study retrospectively examined the prognosis of teeth with root fracture in the maintenance phase of periodontal treatment and assessed the factors influencing the prognosis of teeth with root fracture. The clinical records of 100 patients, who had entered the maintenance phase of periodontal therapy in a university hospital and had experienced fracture of tooth root, were analyzed. The following parameters were included in the evaluation: age and gender of the subjects, tooth type, restoration and root-filling of the teeth with root fracture, and number of teeth present. Survival probability was assessed using Kaplan-Meier analysis. The median survival time (95% confidence interval) was 42 months (22-62 months). Teeth with horizontal fracture had a significantly higher survival probability compared with teeth with vertical fracture. No significant differences were found in the survival probability with regard to gender and tooth type. In conclusion, teeth with horizontal root fracture have a better prognosis compared with teeth with vertical root fracture in patients undergoing periodontal maintenance. PMID- 19302202 TI - Prevalence of sequelae in the permanent anterior teeth after trauma in their predecessors: a longitudinal study of 8 years. AB - The aim of this study was to determine the prevalence of sequelae in the permanent anterior teeth following trauma in their predecessors, and also to verify the existence of association between the sequelae in the permanent teeth and the type of injury in their predecessors according to the age group at the time of injury. This study was performed based on data from the records of 169 boys and 138 girls who received assistance at the dental clinic of the State University of Rio de Janeiro from March 1996 to December 2004. The sample was collected from 753 traumatized deciduous teeth of children aged 0-10 years. The number of boys and girls with dental trauma corresponded to 55.0% and 45.0%, respectively. The more affected age period was between 1 and 4 years (75.3%). The most common type of traumatic injury was the intrusion (29.3%), followed by avulsion (14.1%). Concerning permanent dentition, the most common developmental disturbances were discoloration of enamel and/or enamel hypoplasia (46.08%) and eruption disturbances (17.97%) due to the traumatic injury in their predecessors. It was not possible to find an association between the type of injury in primary teeth and sequelae in their successors in the studied age groups (P 0.05. The most common cause of injuries was falls (49.1%), followed by traffic accidents (13.2%), collision against objects or people (11.3%) and misuse of teeth 9.4%. The commonest type of injury was enamel fracture alone (9.9%), followed by enamel-dentine fracture (4.8%). Majority of the accidents occurred at home (60.4%), followed by school (26.4%). The prevalence of traumatic dental injuries was on the increase among suburban Nigerian children in Ile-Ife and it has a potential to be considered an emerging public health problem. PMID- 19302205 TI - Traumatic tooth injuries to primary teeth of children aged 0-3 years. AB - The purpose of this study was to analyze data according to gender, age, cause, number of traumatized teeth, type of tooth and trauma, from the records of traumatized children (0-3 years) referred to the Ondokuz Mayis University, Dental Faculty, Department of Pedodontics in middle Black Sea region of Turkey. A total of 563 boys and girls participated in the study. Traumatic dental injuries were recorded using the classification of the World Health Organization modified slightly by Andreasen & Andreasen. Traumatic dental injuries were identified in 17.4% of the children. The largest percentage of injuries were in the 13-18 months old children with no significant gender differences (P > 0.05). Periodontal tissue injuries were the most common (84.7%) with no statistical difference between different ages or gender (P > 0.05). The main etiological factor of traumatic dental injury was falls (73.5%) and it was not different between age and gender (P > 0.05). The maxillary central incisors were involved in a higher percentage of traumatic injuries (98%), with no differences between the right and the left sides (P > 0.05). Treatment was sought for 37.4% of children within 1-7 days. It is suggested that parents should be informed about prevention of traumatic injuries and to contact a dentist immediately. PMID- 19302206 TI - Dento-alveolar and maxillofacial injuries among different ethnic groups in Israel. AB - An evaluation of dental and maxillofacial trauma in a level 1 trauma center in Israel was carried out to assess the causes of trauma and the relationship between the injuries to the patient's age, gender and ethnicity. Analysis was based on data from the Israel Trauma Registry. Patients hospitalized in a level 1 trauma center from January 1, 2000 to December 31, 2005 were analyzed according to age, gender, time, place, ethnicity and cause of injury. Files of 22 558 trauma patients were reviewed. Maxillofacial and tooth injuries were separated and further analyzed according to the above parameters. Maxillofacial or dental injuries were observed in 1668 (7.4%) of the trauma patients, caused by motor vehicle accidents (39.2%), falls (30.9%), and intentional injuries (21.2%). Most occurred on the street/road (46.5%), at home (18.8%), and in public buildings (12.4%). Arab patients suffered more from vehicle accidents while Jewish patients presented more intentional injuries. Men were hospitalized three times more than women, and young people were at greater risk. The most frequent age of trauma was 19-28 years (27.6%). Ethnic differences were particularly noticeable for intentional injuries and vehicle accidents. This emphasizes that a larger percentage of the Arab population suffered from dental and maxillofacial injuries. Further dental health education and training for primary caregivers are warranted. PMID- 19302207 TI - Predictive factors for esophageal stenosis after endoscopic submucosal dissection for superficial esophageal cancer. AB - Endoscopic submucosal dissection (ESD) has been utilized as an alternative treatment to endoscopic mucosal resection for superficial esophageal cancer. We aimed to evaluate the complications associated with esophageal ESD and elucidate predictive factors for post-ESD stenosis. The study enrolled a total of 42 lesions of superficial esophageal cancer in 33 consecutive patients who underwent ESD in our department. We retrospectively reviewed ESD-associated complications and comparatively analyzed regional and technical factors between cases with and without post-ESD stenosis. The regional factors included location, endoscopic appearance, longitudinal and circumferential tumor sizes, depth of invasion, and lymphatic and vessel invasion. The technical factors included longitudinal and circumferential sizes of mucosal defects, muscle disclosure and cleavage, perforation, and en bloc resection. Esophageal stenosis was defined when a standard endoscope (9.8 mm in diameter) failed to pass through the stenosis. The results showed no cases of delayed bleeding, three cases of insidious perforation (7.1%), two cases of endoscopically confirmed perforation followed by mediastinitis (4.8%), and seven cases of esophageal stenosis (16.7%). Monovalent analysis indicated that the longitudinal and circumferential sizes of the tumor and mucosal defect were significant predictive factors for post-ESD stenosis (P < 0.005). Receiver operating characteristic analysis showed the highest sensitivity and specificity for a circumferential mucosal defect size of more than 71% (100 and 97.1%, respectively), followed by a circumferential tumor size of more than 59% (85.7 and 97.1%, respectively). It is of note that the success rate of en bloc resection was 95.2%, and balloon dilatation was effective for clinical symptoms in all seven patients with post-ESD stenosis. In conclusion, the most frequent complication with ESD was esophageal stenosis, for which the sizes of the tumor and mucosal defect were significant predictive factors. Although ESD enables large en bloc resection of esophageal cancer, practically, in cases with a lesion more than half of the circumference, great care must be taken because of the high risk of post-ESD stenosis. PMID- 19302208 TI - P53 and Ki-67 overexpression in gastroesophageal reflux disease--Barrett's esophagus and adenocarcinoma sequence. AB - Gastroesophageal reflux disease (GERD) is a major risk factor for the development of esophageal adenocarcinoma (ACE). Many molecular alterations occur in esophageal carcinogenesis, yet the exact mechanism of ACE development remains unknown. This study aims to determine p53 protein and Ki-67 expression in esophageal mucosa of patients with GERD and study the correlation between these markers and the progression from normal squamous epithelium to esophagitis, columnar epithelium with or without intestinal metaplasia and ACE. We analyzed p53 protein and Ki-67 expression in biopsies of 200 patients with GERD and 35 patients with ACE. Those biopsies were classified into five groups: (i) G1 normal squamous epithelium (58); (ii) G2 esophagitis (80); (iii) G3 columnar epitheliums without intestinal metaplasia (30); (iv) G4, columnar epitheliums with intestinal metaplasia (32); and (v) G5 ACEs (35). p53 protein overexpression was found in 7% (4) of G1, 37.5% (30) of G2, 30% (9) of G3, 62.5% (20) of G4, and 71.4% (25) of G5 (p < 0.001). Ki-67 index increased according to the severity of histopathological diagnoses. Ki67 index was 21.3 +/- 19.5% in G1, 38.8 +/- 24.9% in G2, 37.7 +/- 26.3% in G3, 52.8 +/- 24.6% in G4, and 57.1 +/- 25.1% in G5 (P < 0.001). Linear correlation between p53/Ki67 expression and the multistep progression from squamous epithelium to ACE was observed (P < 0.001 and P < 0.05). Our results indicate that overexpression of p53 and increased Ki-67 could be associated with the development and progression to ACE in patients with GERD. PMID- 19302209 TI - Endocytoscopic observation for esophageal squamous cell carcinoma: can biopsy histology be omitted? AB - We examined whether endocytoscopic observation of esophageal squamous cell carcinoma can replace the histologic examination of biopsy specimens. In a basic investigation, we examined 57 iodine-unstained areas in the resected specimens of the esophagus from 28 individuals. The endocytoscopic findings were graded from 0 to 3 in tandem with observations of the iodine staining. For endocytoscopic observation, we sprayed 1% methylene blue or toluidine blue as a vital dye on the surface of the esophageal mucosa, allowing 15-20 s for sufficient staining. One endoscopist observed the target lesions and decided their endocytoscopic type classification. Histological diagnoses were made by two pathologists who were unaware of the endoscopic findings. We then compared the endocytoscopic diagnosis and conventional histological diagnosis. In an in vivo investigation, we examined 71 lesions of esophageal squamous cell carcinoma. Two endoscopists diagnosed the type classification in consultation with a pathologist with regard to 'nuclear density,''nuclear abnormality,' and 'whether biopsy histology could have been omitted on the basis of endocytoscopic findings.' For the in vivo observation, we utilized XEC120U (higher magnification type [x1100]), XEC300F (lower magnification type [x450]), and XGIF-Q260EC1 (lower magnification type [x450]) instruments. In the basic investigation, among the 11 areas classified as Type 1, 10 (91%) were category 1 by the Vienna classification. Among the 39 lesions classified as Type 3, 36 (92%) were category 4 or 5. The sensitivity of endocytoscopy for malignant lesions (Vienna classification categories 4 and 5) was 94.7%, if Type 3 was considered malignant. The specificity was 84.2% according to the same criteria. In the in vivo observation, two endoscopists diagnosed more than 90% of esophageal squamous cell carcinomas as neoplasms using each type of endocytoscope. With regard to nuclear density, the pathologist considered it to be increased in 98% of cases with the XEC120U, in 94% with the XEC300F, and in 93% with the XGIF-Q260EC1. With regard to nuclear abnormality, the positivity rate was 90% with the XEC120U, 78% with the XEC300F, and 80% with the XGIF-Q260EC1. As to whether or not biopsy histology examination was considered necessary, the pathologist made a 'Yes' judgment for 84% of cases observed with the XEC120U, 66% with the XEC300F, and 67% with the XGIF-Q260EC1. Cancerous lesions diagnosed as Type 3 by both endoscopists using the XEC120U accounted for 46 (90.2%) of the 51 cases. Among these 46 cases, biopsy histology was considered unnecessary by the pathologist in 43 (93.5%). We believe that endocytoscopic observation has the potential to reduce the extent of histologic examination of biopsy specimens in cases corresponding to Types 1 and 3 of our classification. PMID- 19302210 TI - Targets for molecular therapy in esophageal squamous cell carcinoma: an immunohistochemical analysis. AB - Neoadjuvant chemotherapy may improve the outcome of esophageal cancer after esophagectomy, but is accompanied by considerable toxicity by collateral destruction of normal cells. Such side effects may be avoided by developing therapies that specifically target molecular characteristics of tumors. The aim of the present study was to determine the proportion of esophageal squamous cell carcinoma (ESCC) patients that could possibly benefit from (a combination of) currently available targeted therapies, by assessing the frequency of immunohistochemical expression of their target molecular markers in ESCC tissues. Sections from a validated tissue microarray comprising 108 ESCCs were immunohistochemically stained for Bcl-2, c-KIT, cyclo-oxygenase-2 (COX-2), cyclin D1, estrogen receptor (ER), epidermal growth factor receptor (EGFR), Her-2/neu, progesterone receptor (PR), and vascular endothelial growth factor (VEGF). VEGF, cyclin D1, EGFR, and COX-2 could be detected in 55, 42, 40, and 40%, respectively. Her-2/neu, Bcl-2, and c-KIT were detected in 12, 11, and 10% of the tumors, respectively. No nuclear expression of ER or PR was noticed. Concurrent expression of two markers was noticed in 28% of ESCCs, whereas 25% of ESCCs showed concurrent expression of three markers. The concurrent expression of two of the most frequently expressed markers (VEGF, cyclin D1, EGFR, and COX-2) ranged from 11 (COX-2 and EGFR) to 26% (cyclin D1 and VEGF). The expression of all of these four markers was seen in 5% of ESCCs. Promising targets for molecular therapy in ESCC appear to be COX-2, VEGF, EGFR, and cyclin D1, as they are frequently overexpressed. Phase II clinical studies on these molecular markers may therefore be warranted. The role for targeted therapy against ER, PR, Her-2/neu, c-KIT, or Bcl-2 in ESCC seems limited. PMID- 19302211 TI - Comparative study of different treatments for malignant tracheoesophageal/bronchoesophageal fistulae. AB - The aim of this study is to compare the survival time and quality of life (QOL) of patients who have received different treatment for tracheoesophageal/bronchoesophageal fistula. Between January 2003 and December 2007, 35 patients with malignant tracheoesophageal/bronchoesophageal fistula were recorded as the control group, gastrostomy group, and stenting group, respectively, according to the treatments they chose. Two weeks after the treatment, European Organization for Research and Treatment of Cancer Quality of Life Core 30 Questionnaire (QLQ-C30), Quality of Life Questionnaire-esophageal module (QLQ-OES18), and a respiratory symptom-related QOL index are employed to assess QOL of these patients. There is no significant difference in survival time and constituent ratio of death reason among groups. Except for eight patients who died within 2 weeks after the treatment, all other 27 patients returned back the questionnaires. As compared to the control group, patients in the gastrostomy group gained a low score in emotional function and financial situation, while patients in the stenting group had lower scores in financial problems and seven respiratory and eating-related symptoms. In contrast with the gastrostomy group, patients in stenting group had higher scores in emotional and social functions, and lower scores in six respiratory and eating-related symptoms. With patients' QOL considered, the self-expandable coated stenting should be the first choice of therapy for malignant tracheoesophageal/bronchoesophageal fistula, whereas gastrostomy should be kept from use. PMID- 19302212 TI - Prevalence of esophageal dysmotility in a cohort of patients with esophageal biopsies consistent with eosinophilic esophagitis. AB - Eosinophilic esophagitis (EoE) is increasingly being diagnosed in adults presenting with dysphagia, food impactions, and chest pain. Studies to date provide conflicting data on the association of EoE and esophageal dysmotility. The objective of this study was to evaluate the prevalence of esophageal dysmotility in a cohort of patients with biopsies consistent with EoE at a military treatment facility. This is a prospective evaluation of consecutively identified patients at our institution diagnosed with EoE from March 1, 2005 to June 1, 2007. Thirty-two patients with biopsies consistent with EoE completed a symptom survey and 30 underwent esophageal manometry. The majority of EoE patients (23/30, 77%) had a normal end-expiratory lower esophageal sphincter (LES) pressure (normal range 10-35), whereas six patients had a low-normal LES pressure (6-9 mm Hg) and one patient had a decreased LES pressure (<5 mm Hg). Five patients (15.6%) were diagnosed with a nonspecific esophageal motor disorder (NSEMD). Two patients had high mean esophageal amplitude contractions >180 mm Hg (188 mm Hg, 209 mm Hg). No patient was diagnosed with nutcracker esophagus or diffuse esophageal spasm. Patients with and without NSEMD reported a similar degree of swallowing difficulty, heartburn, belching, chest pain, regurgitation, symptoms at night, and total symptom score. Likewise, eosinophil count on mucosal biopsy was similar between patients with and without a NSEMD. In this cohort, we found the prevalence of an NSEMD to be similar to that of a 10% prevalence found in a gastroesophageal reflux population. PMID- 19302213 TI - Outcome of palliative esophageal stenting for malignant dysphagia: a retrospective analysis. AB - Greater than 50% of patients with esophageal carcinoma are found to be incurable at the time of diagnosis, leaving only palliative options. Self-expanding metal stents (SEMs) are effective for relieving symptoms and complications associated with esophageal carcinoma and improving quality of life. We undertook a retrospective analysis to evaluate the experience of palliative esophageal stenting for symptomatic malignant dysphagia in our institution over a period of 7 years. Between January 1999 and January 2006, 126 patients who received SEMs for malignant dysphagia were identified using an upper gastrointestinal specialist nurse clinician database. Data were obtained from patient case notes, endoscopy, histopathology, radiology, and external agency databases. Of the 126 identified, 36 patients were excluded from the analysis. A number of variables including age, sex, presenting complaints, type of stent, indications of stenting, success or failure of stent insertion, survival rate, and complication rate were analyzed. Of the 90 patients, 55 (61%) were male and 35 (39%) were female. The mean age of patients was 70.79 (range 40-97) years. The predominant presenting complaints were dysphagia (n = 81) and weight loss (n = 48). The indication for stenting was worsening dysphagia in all patients. Tumors were confined to the distal esophagus and esophagogastric junction in 73 patients (81%), and the mid-esophagus in 17 (19%). Adenocarcinoma was identified in 61 patients (67.8%) and squamous cell carcinoma in 29 (32.2%). Stenting numbers were comparable in endoscopic and radiologic groups (47 vs. 43), with successful stent deployment in 89 patients. The 7- and 30-day mortality was 9% (n = 8) and 28% (n = 25), respectively. Comparable numbers of early deaths were seen in both radiologic (n = 13) and endoscopic (n = 12) groups. Causes of early inpatient death included hemorrhage (n = 5), pneumonia (n = 7), exhaustion (n = 2), cardiac causes (n = 3), perforation (n = 1), and sepsis (n = 1). The number of patients with complications was 41 (45.6%), 25 in the surgical group and 15 in the radiologic group; the difference was not significant (P = 0.13). The mean survival time was 92.5 (0-638) days and median survival time was 61 days. A subgroup of patients with complete dysphagia (score 4) gained a mean survival of 59 days. Those patients receiving adjuvant chemotherapy or radiotherapy survived significantly longer than those receiving stenting alone (152.8 days vs. 71.8 days). There is no significant difference in complications or survival when using endoscopic or radiologic methods to deploy SEMs in patients with inoperable esophageal cancer. Mortality is low; however, the morbidity rate is significant. Patients receiving adjuvant chemotherapy or radiotherapy, in addition to stenting, survived significantly longer than those with a stent only. PMID- 19302214 TI - Phagocytosis of bacteria is enhanced in macrophages undergoing nutrient deprivation. AB - Phagocytosis represents a mechanism used by macrophages to remove pathogens and cellular debris. Recent evidence suggests that phagocytosis is stimulated under specific conditions of stress, such as extracellular pressure and hypoxia. In the present study, we show that amino acid or glucose deprivation caused an increase in the phagocytosis of heat-inactivated Escherichia coli and Staphylococcus aureus by macrophages, but not the uptake of platelets, apoptotic cells or beads. Increased phagocytosis of bacteria could be blocked by phagocytosis inhibitors and was found to be dependent on p38 mitogen-activated protein kinase activity and scavenger receptor A. Although nutrient deprivation is a strong stimulus of autophagy, autophagosome formation was not critical for the uptake of bacteria because phagocytic clearance was not inhibited after down-regulation of the autophagy essential gene Atg7. Moreover, enhanced uptake of bacteria should not be considered as a general stress response because phagocytosis of bacteria was not stimulated after exposure of macrophages to the genotoxic agent camptothecin, heat (40 degrees C) or thapsigargin-induced endoplasmic reticulum stress. Overall, the results obtained in the present study indicate that nutrient deprivation can stimulate macrophages to fight bacterial infections. PMID- 19302215 TI - Probing the substrate specificities of matriptase, matriptase-2, hepsin and DESC1 with internally quenched fluorescent peptides. AB - Type II transmembrane serine proteases are an emerging class of proteolytic enzymes involved in tissue homeostasis and a number of human disorders such as cancer. To better define the biochemical functions of a subset of these proteases, we compared the enzymatic properties of matriptase, matriptase-2, hepsin and DESC1 using a series of internally quenched fluorogenic peptide substrates containing o-aminobenzoyl and 3-nitro-tyrosine. We based the sequence of the peptides on the P4 to P4' activation sequence of matriptase (RQAR-VVGG). Positions P4, P3, P2 and P1' were substituted with nonpolar (Ala, Leu), aromatic (Tyr), acid (Glu) and basic (Arg) amino acids, whereas P1 was fixed to Arg. Of the four type II transmembrane serine proteases studied, matriptase-2 was the most promiscuous, and matriptase was the most discriminating, with a distinct specificity for Arg residues at P4, P3 and P2. DESC1 had a preference similar to that of matriptase, but with a propensity for small nonpolar amino acids (Ala) at P1'. Hepsin shared similarities with matriptase and DESC1, but was markedly more permissive at P2. Matriptase-2 manifested broader specificities, as well as substrate inhibition, for selective internally quenched fluorescent substrates. Lastly, we found that antithrombin III has robust inhibitory properties toward matriptase, matriptase-2, hepsin and DESC1, whereas plasminogen activator inhibitor-1 and alpha(2)-antiplasmin inhibited matriptase-2, hepsin and DESC1, and to a much lesser extent, matriptase. In summary, our studies revealed that these enzymes have distinct substrate preferences. PMID- 19302216 TI - The regulation of the endosomal compartment by p53 the tumor suppressor gene. AB - The endosomal compartment of the cell is involved in a number of functions including: (a) internalizing membrane proteins to multivesicular bodies and lysosomes; (b) producing vesicles that are secreted from the cell (exosomes); and (c) generating autophagic vesicles that, especially in times of nutrient deprivation, supply cytoplasmic components to the lysosome for degradation and recycling of nutrients. The p53 protein responds to various stress signals by initiating a transcriptional program that restores cellular homeostasis and prevents the accumulation of errors in a cell. As part of this process, p53 regulates the transcription of a set of genes encoding proteins that populate the endosomal compartment and impact upon each of these endosomal functions. Here, we demonstrate that p53 regulates transcription of the genes TSAP6 and CHMP4C, which enhance exosome production, and CAV1 and CHMP4C, which produce a more rapid endosomal clearance of the epidermal growth factor receptor from the plasma membrane. Each of these p53-regulated endosomal functions results in the slowing of cell growth and division, the utilization of catabolic resources and cell-to cell communication by exosomes after a stress signal is detected by the p53 protein. These processes avoid errors during stress and restore homeostasis once the stress is resolved. PMID- 19302217 TI - Nonspecific esophageal motility disorders may be an early stage of a specific disorder, particularly achalasia. AB - The clinical significance of nonspecific esophageal motility disorder (NEMD) is unclear. Our aim was to investigate the natural history of NEMD. All manometries performed at Meir Hospital from 1997 to 2004 and diagnosed as NEMD were reviewed. Manometric criteria for NEMD included either low-amplitude peristalsis, nonprogression of peristalsis, prolonged retrograde or triple-peaked waves, or incomplete relaxation of the lower sphincter. Patients determined to have NEMD were contacted and asked to complete a questionnaire and undergo a second manometry. NEMD had been diagnosed in 137 patients. Upon review of manometry results, 65 patients were eligible for the study (36 men and 29 women). The other 72 patients did not have NEMD when we reviewed their manometry tracing, applying strict criteria as specified in Table 1. The average age was 64 +/- 16 years (range 24-83 years). The average follow-up period was 7 +/- 2 years. All 65 patients were symptomatic at their initial prestudy visit. By the second visit, symptoms had resolved in 33 (51%) patients and improved in 13 (19%). Dysphagia, chest pain, and food regurgitation had improved, whereas heartburn and respiratory symptoms had not. Of 37 patients with triple-peaked waves, only 11 (30%) had improved clinically. Of the 65 study patients, 17 (26%) had a second manometry during the study, which was normal in 2 (12%), unchanged in 11 (69%), and revealed achalasia in 4 (23%), representing 6% of all study patients. NEMD is generally a benign disorder that improves clinically in most cases. Nevertheless, in about 6% of patients, NEMD may evolve into achalasia. PMID- 19302218 TI - Gastric secretory and hormonal patterns in end-stage chagasic achalasia. AB - Achalasia surgical treatment alters the esophagogastric junction anatomy (cardiomyotomy plus fundoplication or esophagectomy and gastric pull-up), thus favoring a certain degree of gastroesophageal reflux. Gastric secretory and hormonal functioning is not completely known in chagasic patients. The aim of this study was to evaluate the gastric secretory and hormonal response in patients with end-stage chagasic achalasia compared with normal subjects. Gastric secretion and hormonal response were assessed by estimation of gastric acid secretion (GAS) in basal condition and after pentagastrin stimulation, basal serum gastrin, and serum pepsinogen (SP) in basal condition and after betazole hydrochloride (Histalog; Eli Lilly and Company, Indianapolis, IN, USA) stimulation in 27 patients with chagasic achalasia. The results were then compared with those of 24 normal subjects. In the chagasic group, the mean basal and stimulated GAS were significantly lower than in the control group (basal: 1.277 vs. 3.13, P = 0.002; stimulated: 15.9 vs. 35.8, P = 0.0001). Chagasic patients' SG levels showed a significantly higher basal value than the control group (83.3 vs. 36.8, P = 0.0001). There was a significant increase of SP after stimulation compared with the basal levels in both chagasic and control groups. Although the chagasic patients' SP values were higher than the controls, this difference was not statistically significant, either in basal and stimulated conditions (basal: 122.0 vs. 108.9, stimulated 120 min: 177.1 vs. 158.9). In patients with chronic Chagas' disease (ChD), although autonomic denervation does not suppress the strength of the gastric mucosal cells' secretory response to stimulation, it reduces GAS (parietal cell) without, however, affecting SP production (chief cells). On the other hand, the gastrin-producing cells have continuously been stimulated by low GAS. PMID- 19302219 TI - p53 Arg72Pro, MDM2 T309G and CCND1 G870A polymorphisms are not associated with susceptibility to esophageal adenocarcinoma. AB - p53 Arg72Pro, MDM2 T309G, and CCND1 G870A are functional single-nucleotide polymorphisms (SNPs) in key genes that regulate apoptosis and cell cycle. Variant genotypes of these SNPs have been associated with increased risk and earlier age of onset in some cancers. We investigated the association of these SNPs with susceptibility to esophageal adenocarcinoma in a large, North American case control study. Three hundred and twelve cases and 454 cancer-free controls recruited in Boston, USA were genotyped for each of the three SNPs, and demographic and clinical data were collected. Genotype frequencies for each of the three SNPs did not deviate from the Hardy-Weinberg equilibrium, and did not differ between cases and controls. Odds ratios (OR), adjusted for clinical risk factors, for the homozygous variant genotypes were 0.99 (95% confidence interval [CI] 0.57-1.72) for p53 Pro/Pro, 0.81 (95% CI 0.52-1.28) for MDM2 G/G, and 0.97 (95% CI 0.64-1.49) for CCND1 A/A. The analysis was adequately powered (80%) to detect ORs of 1.37, 1.35, and 1.34 for each SNP, respectively. In contrast to the results of smaller published studies, no association between p53 Arg72Pro, MDM2 T309G, and CCND1 G870A SNPs and susceptibility to esophageal adenocarcinoma, age of onset, or stage of disease at diagnosis was detected. PMID- 19302220 TI - Thirty-four cases of esophageal perforation: the experience of a district general hospital in the UK. AB - Esophageal perforation is uncommon and traditionally has a high rate of morbidity and mortality. Our aim was to perform a 13-year retrospective review of the cases managed in our district general hospital. Thirty-four cases of esophageal perforation diagnosed between 1995 and 2008 were retrospectively analyzed. There were 20 males and 14 females with a median age of 64 (range 23-86) years. The etiology of the perforations were Boerhaave's syndrome (n= 19), therapeutic endoscopy (n= 9), diagnostic endoscopy (n= 2), gastric lavage injury (n= 1), foreign body (n= 1), blunt chest trauma (n= 1), and spontaneous tumor perforation (n= 1). Only 11 cases (32%) had evidence of surgical emphysema upon examination. In 50% of cases, another clinical diagnosis was initially suspected. Twenty-four were treated surgically and 10 cases managed non-operatively. Surgical treatment included thoracotomy with primary repair (n= 9), T-tube drainage (n= 7), emergency esophagectomy (n= 1), or intra-operative stent insertion (n= 1). Four cases had primary repair and fundal wrap via abdominal approach without thoracotomy. Two patients were treated with washout and drainage only. Eight patients died overall (in-hospital mortality 23.5%). Esophageal perforations are often initially misdiagnosed and the majority do not have surgical emphysema. There are a wide variety of methods to manage esophageal perforation. Management tailored to the location and size of perforation, degree of contamination, and underlying cause appears to result in a reasonable prognosis. PMID- 19302221 TI - Intraluminal brachytherapy in the management of squamous carcinoma of the esophagus. AB - Intraluminal high dose rate brachytherapy (ILHDR BT) is one of several effective modalities for palliation of advanced esophageal cancer. Thirty patients with endoscopic-proven, mostly locally advanced, squamous cell carcinoma of the esophagus, not involving the gastroesophageal junction and without distant metastases, were included in this analysis. Twenty-nine patients received two ILHDR BT sessions of 8 Gy within a week and one patient received only one session. All patients were followed monthly. Outcomes included quality of life (QOL), symptoms control: dysphagia, regurgitation, odynophagia, and chest or back pain, as well as, overall survival. Through 4 months of follow-up, QOL was statistically improved (having lowered scores) in regards to feelings (P= 0.013), sleeping (P= 0.032), eating (P= 0.020), and social life (P= 0.002). The most significantly improved symptom was dysphagia (P < 0.006), with a reduction of 0.52 units or one-half grade. Regurgitation, odynophagia, and pain were lower during follow-up but were not statistically significant. The median overall survival from death of any cause was 165 days (with a 95% confidence interval of 128-195 days). In conclusion, ILHDR BT of advanced squamous esophageal cancer consisting of two out-patient procedures is very successful in achieving the primary objectives of the patients to reduce dysphagia and improve QOL. PMID- 19302222 TI - HIF-1alpha protein expression is associated with the environmental inflammatory reaction in Barrett's metaplasia. AB - The oxygen-regulated transcription factor subunit hypoxia inducible factor-1alpha (HIF-1alpha) is involved in angiogenesis, energy metabolism, cell survival, and inflammation. We examined the protein expression of HIF-1alpha within the progression of Barrett's sequence as well as the type and degree of the environmental inflammatory reaction. Squamous epithelium (SE), metaplastic, low- and high-grade dysplastic lesions, and tumor tissue of 57 resection specimens from patients with Barrett's adenocarcinoma were immunohistochemically analyzed. Active and chronic inflammatory reactions were classified according to the Updated Sydney System. HIF-1alpha protein expression increased significantly from SE to Barrett's metaplasia (BM) (P < 0.0001). From metaplasia through low- and high-grade dysplasia to cancer, no further increase could be detected. Active and chronic inflammation were also significantly different between SE and BM (P < 0.0001) but not during further progression in the sequence. HIF-1alpha protein expression did not correlate with histopathologic parameters or survival. HIF 1alpha protein expression pattern resembles the active and chronic environmental inflammatory reaction. All were significantly increased in metaplasia compared to SE without further change in tumor development. HIF-1alpha protein expression appears to be associated with inflammatory processes in the development of BM. PMID- 19302223 TI - When did the esophagus start shrinking? The history of the short esophagus. AB - Even though the history of this condition extends for almost 100 years, the short esophagus (SE) is still one of the most controversial topics in esophageal surgery with its existence still denied by some distinguished surgeons. We reviewed the evolution behind the diagnosis and treatment of the SE and the persons who wrote its history, from the first descriptions by radiologists, endoscopists, and surgeons to modern treatment. PMID- 19302224 TI - Salvage esophagectomy after exclusive chemoradiotherapy: results at the Brazilian National Cancer Institute (INCA). AB - Surgical resection is considered the gold standard treatment for esophageal cancer, with global cure rates ranging from 15 to 40%. Exclusive chemoradiotherapy has been used for patients with locally advanced esophageal carcinoma or without clinical conditions for esophagectomy, reaching a 5-year survival rate of up to 30%. However, locoregional control is poor, with local recurrence of 40-60%, being reported in the literature. Maybe, these patients can benefit from salvage surgery. In this study, 15 patients with esophageal cancer submitted to salvage esophagectomy after exclusive chemoradiotherapy treatment were retrospectively analyzed. Salvage esophagectomy was demonstrated to be technically feasible. However, it presents with high surgical morbidity. Currently, salvage esophagectomy is considered the best available treatment to attempt cure in cases of tumor recurrence or persistence after exclusive chemoradiotherapy. All the other types of treatments are regarded as palliative with discouraging survival results. PMID- 19302225 TI - Effects of detraining on muscle strength and mass after high or moderate intensity of resistance training in older adults. AB - This study examined the effects of a 12 weeks detraining period on muscle strength and mass in older adults who had previously participated in a 12 weeks resistance training programme of high [80% of one repetition maximum (1-RM)] or moderate (60% of 1-RM) intensity. Twenty older adults (60-74 years), separated into a high (HI; n = 10; age: 65 +/- 5 years) and a moderate (MI; n = 10; age: 66 +/- 4 years) intensity resistance training group, were measured in the 1-RM knee extension and flexion strength, and the midthigh cross sectional areas (CSAs) of quadriceps, hamstrings and total thigh before and after a 12 weeks training period as well as after a 12 weeks detraining period. Maximum knee extension and flexion strength and the CSAs of all muscles decreased significantly (P<0.05) with detraining but remained higher (P<0.05) than pretraining levels for both groups. The HI group had a greater decrement (P<0.05) in maximum strength and the CSA of total thigh compared to the MI group but strength levels and the CSA following detraining were higher (P<0.05) for the HI group. The above data suggest that after a short detraining period of 12 weeks, muscle strength and hypertrophy levels of older adults decrease but remain greater than pretraining irrespective of training intensity. Greater declines in muscle strength are observed following HI training but still muscular strength and muscle mass are retained at a higher level than with MI probably due to the higher gains achieved during the training period. PMID- 19302226 TI - Cardiac magnetic resonance imaging in valvular heart disease. AB - Cardiac magnetic resonance imaging (CMR) has rapidly gained acceptance as an accurate, reproducible and non-invasive imaging method for assessment of a wide range of cardiovascular diseases. However, CMR has not been used widely for diagnostic purposes in valvular heart disease (VHD). Unlike echocardiography it has no body habitus-related limitations and can thus be used to complement echocardiography. It is an especially good alternative for clinical follow-up in patients with VHD, as it allows accurate measurement of valvular dysfunction and related ventricular burden. Additionally, CMR is an ideal method for evaluating complex congenital heart disease and determining the significance of its components. It can also be used to study the physiological course of valvular dysfunction and response to therapeutic interventions. In this review, we present a basic introduction to CMR methodology, including its advantages and potential problems, and the physiology and quantification in VHD. We also discuss clinical applications of CMR in VHD. Furthermore, we describe how a CMR study statement should be structured in order to increase clinical use of this valuable methodology in cardiology. PMID- 19302227 TI - Inert gas rebreathing: the effect of haemoglobin based pulmonary shunt flow correction on the accuracy of cardiac output measurements in clinical practice. AB - BACKGROUND: Cardiac output (CO) is an important cardiac parameter, however its determination is difficult in clinical routine. Non-invasive inert gas rebreathing (IGR) measurements yielded promising results in recent studies. It directly measures pulmonary blood flow (PBF) which equals CO in absence of significant pulmonary shunt flow (Q(S)). A reliable shunt correction requiring the haemoglobin concentration (c(Hb)) as only value to be entered manually has been implemented. Therefore, the aim of the study was to evaluate the effect of various approaches to Q(S) correction on the accuracy of IGR. METHODS: Cardiac output determined by cardiac magnetic resonance imaging (CMR) served as reference values. The data was analysed in four groups: PBF without correcting for Q(S) (group A), shunt correction using the patients' individual c(Hb) values (group B), a fixed standard c(Hb) of 14.0 g dl(-1) (group C) and a gender-adapted standard c(Hb) for male (15.0 g dl(-1)) and female (13.5 g dl(-1)) probands each (group D). RESULTS: 147 patients were analysed. Mean CO(CMR) was 5.2 +/- 1.4 l min(-1), mean CO(IGR) was 4.8 +/- 1.3 l min(-1) in group A, 5.1 +/- 1.3 in group B, 5.1 +/- 1.3 l min(-1) in group C and 5.1 +/- 1.4 l min(-1) in group D. The accuracy in group A (mean bias 0.5 +/- 1.1 l min(-1)) was significantly lower as compared to groups B, C and D (0.1 +/- 1.1 l min(-1); P<0.01). CONCLUSION: IGR allows a reliable non-invasive determination of CO. Since PBF significantly increased the measurement bias, shunt correction should always be applied. A fixed c(Hb) of 14.0 g dl(-1) can be used for both genders if the exact c(Hb) value is not known. Nevertheless, the individual value should be used if any possible. PMID- 19302228 TI - Methicillin resistant Staphylococcus aureus (MRSA): Diagnostic, clinical relevance and therapy. AB - In the last decades, increasing numbers of patients with problematic bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) have been reported. Beside the common problem of MRSA variants in hospitals, recently community-based MRSA (cMRSA) has become a growing problem even in patients without typical risk factors. cMRSA often carries the virulence factor Panton-Valentine-leukocidin (PVL) causing dermatologic diseases like therapy-refractory furunculosis in young adults. Thus, it is both a medical and health economic issue to identify MRSA as quickly as possible and then eradiate it completely. We review the practical consequences and in particular, the therapy options which are reasonable once MRSA is identified. PMID- 19302229 TI - Diseases caused by human papillomaviruses (HPV). AB - Human papillomaviruses (HPV) are non-enveloped tumor viruses with a double stranded DNA approximately 8 kilobases in length. The viral genome is enclosed by a spherical capsid with icosahedral symmetry and a diameter of about 55 nm. More than 100 HPV types have been identified. They infect the squamous epithelia of skin and mucosa and usually cause benign papillomas or warts. Persistent infection with high-risk oncogenic HPV causes all cervical cancers, most anal cancers, and a subset of vulvar, vaginal, penile and oropharyngeal cancers. In recent years cutaneous beta-HPV types have been associated with the pathogenesis of non-melanoma skin cancers. Two prophylactic HPV vaccines based on virus-like particles (VLP) are licensed. These are up to 100% effective in preventing HPV 16 and HPV 18 infections and associated genital lesions in women, who have not been previously infected with these types. One vaccine also prevents genital warts caused by HPV 6 and HPV 11. PMID- 19302230 TI - Test-retest reliability of the IDEEA system in the quantification of step parameters during walking and stair climbing. AB - The aim of this investigation was to evaluate the inter- and intra-session reliability of spatio-temporal gait variables collected during walking and stair climbing with the Intelligent Device for Energy Expenditure and physical Activity (IDEEA) accelerometer-based system. Eighteen healthy subjects (10 men, 8 women) completed a standardized indoor circuit comprised of walking and stair climbing. Intra-and inter-session reliability was investigated for several pertinent spatio temporal gait variables using intraclass correlations [ICC (3,1)]. Intra-session reliability during walking showed a high reliability of the IDEEA with ICCs ranging between 0.84 (number of steps) and 0.97 (single limb support/double limb support). The ICCs for stair climbing were slightly lower than those during walking with values ranging between 0.74 (step duration) and 0.92 (number of steps). Inter-session reliability during walking showed a high reliability of the IDEEA between all trials, with values ranging between 0.87 (speed) to 0.98 (step duration). The ICCs of stair climbing were again lower than those during walking with values ranging from 0.64 (swing duration) to 0.79 (number of steps). The IDEEA accelerometer-based system provided a highly reliable measurement of spatio temporal variables, in healthy subjects, during walking with moderately reduced correlations during stair climbing. PMID- 19302231 TI - Maximal explosive muscle power in obese and non-obese prepubertal children. AB - OBJECTIVE: The objectives of the present study was to compare the maximal explosive muscle power developed by the lower limbs in obese and non-obese prepubertal children. DESIGN: Twenty-five obese [mean body mass index (BMI) z score: 2.4] and 30 non-obese (mean BMI z-score: 0) children aged 8-12 years, participated in this study. Body composition was measured by bioelectrical impedance analysis and the maximal explosive power of the lower limbs was assessed by the Exercise-Ergometer (a sledge dynamometer). RESULTS: Absolute peak force (F(peak)) was higher in obese than in non-obese children by 18.2% (P<0.001). Peak speed (v(peak)) was not significantly different between groups (P = 0.504). Consequently, absolute peak power (W'(peak)) was higher in obese than in non-obese children (+19.3%, P = 0.021). Considering gender differences, absolute F(peak) and v(peak) were higher in boys than in girls by +12.3 and +17.4% (P<0.05), respectively, thus yielding higher absolute W'(peak) values in boys as compared to girls (+26.1%, P<0.001). Finally, W'(peak) normalized for fat free mass (FFM) was not different between obese and non-obese children but higher in boys than in girls (+24.5%, P<0.001). CONCLUSIONS: Power generation capability per unit of FFM was similar between obese and non-obese children but was significantly higher in boys than girls. On the clinical practice it seems important to devote supplementary care to sustain and improve the motor function of obese and non-obese girls. PMID- 19302232 TI - Enhanced expression of CD71, transferrin receptor, on immature reticulocytes in patients with paroxysmal nocturnal hemoglobinuria. AB - Erythroid cells in the marrow express CD71, transferrin receptor, and reticulocytes released from the marrow lose their expression during maturation. The immature reticulocyte fraction, the proportion of reticulocytes with the highest content of RNA, has been determined by hematology analysis. In the present study, we examined CD71 expression on immature reticulocytes by flow cytometry (FCM) in paroxysmal nocturnal hemoglobinuria (PNH) patients with reticulocytosis. We modified 'reticulocyte-gated FCM' to multi-color FCM, i.e. RNA/CD71, RNA-CD59 or CD59/CD71/CD45. In PNH, in addition to the increased number of immature reticulocytes (%CD71-positive), a more immature phenotype in regard to both CD71 intensity and RNA content levels was demonstrated. In seven PNH patients studied, %CD71-positive reticulocytes were significantly increased at 32.2 +/- 11.9% (n = 10, normal 10.4 +/- 3.5%, P = 0.002). RNA content levels (assessed by mean fluorescence index, MFI) in CD71-positive reticulocytes were significantly increased at 812.0 +/- 215.2 MFI in PNH (n = 10, normal 508 +/- 86.1 MFI, P = 0.002). These data indicate that stimulated erythropoietic conditions induce the release of more immature reticulocytes to the peripheral blood than ordinary erythropoietic conditions. CD71 intensity on immature reticulocytes was well correlated with their RNA content levels, indicating the usefulness of CD71 as an immature reticulocyte marker. PMID- 19302233 TI - Can Thromboelastography performed on kaolin-activated citrated samples from critically ill patients provide stable and consistent parameters? AB - Thromboelastography (TEG) is a potentially useful tool but analysis within 4-6 min of collection imposes limitations on its use and access. The use of citrate blood tubes potentially increases the time frame for processing specimens. There is, however, limited research on the stability of citrate specimens, timing of processing and the accuracy of TEG results. The purpose of this study was to examine the effects of early and delayed processing on TEG parameters using kaolin-activated citrated blood samples in the intensive care population. TEG analysis was performed on 61 patients. Blood was collected into two 3.2% sodium citrate (0.105 m) tubes. Kaolin-activated samples were analysed at 15, 30 and 120 min postcollection. TEG parameters analysed included reaction time (R), clot formation time (K), alpha angle (alpha), maximum amplitude, LY30, the coagulation index, time to maximum rate of thrombus generation, maximum rate of thrombus generation and total thrombus generation. Sixty-one critically ill patients were included. The results of the anova showed that time from collection was significantly associated with the TEG((R)) results (P < 0.05). On comparison of individual outcome variables, this difference in most cases was due to changes over time from 30 to 120 min. Furthermore, progressive changes in TEG parameters such as decreasing R were suggestive of a trend toward hypercoagulability of the specimens. Processing of kaolin-activated citrate TEG specimens can begin as early as 15 min postvenipuncture. However, delaying processing by more than 30 min leads to a significant change in results. PMID- 19302234 TI - Expression profile of Notch-related genes in multidrug resistant K562/A02 cells compared with parental K562 cells. AB - Drug resistance is the major setback of acute myeloid leukemia (AML) therapy. Notch proteins have demonstrated functional regulation in cell proliferation, differentiation, and apoptosis and thus may affect drug resistance. Our study aimed to identify the Notch-related gene profile in drug-resistant AML cells and provide potential strategies for resistant AML therapy. 3-(4,5-Dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide assay was conducted to detect the cytotoxicity of adriamycin toward K562 and drug-resistant K562/A02. Intracellular mean fluorescence intensity was monitored to reflect the intake of adriamycin by confocal microscopy. cDNA microarray was used to test the expression of 113 Notch signaling pathway-related genes in K562/A02 and K562. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) and western blot were used to validate the results from microarray. K562/A02 cells showed a 65-fold higher IC(50) to adriamycin and less intracellular accumulation of adriamycin than K562. cDNA microarray showed marked increases in binding of collagen and cell proliferation-related genes (CD44, DLL3, IL17B, NUMB, and NUMBL) and decreases in signal transduction and transcription factor activity related genes (FZD9, GBP2, GLI1, GLI2, IFNG, KRT5, Notch2, and Notch3). The change of gene expression was further validated by real-time RT-PCR and western blot. Notch signaling pathway related genes may contribute to the drug resistance of AML. PMID- 19302235 TI - Apoptosis inducing and differentiation enhancement effect of oridonin on the all trans-retinoic acid-sensitive and -resistant acute promyelocytic leukemia cells. AB - We investigated the effects of oridonin (Ori), a diterpenoid isolated from Rabdosia rubescens, on apoptosis and differentiation of all-trans-retinoic acid (ATRA)-sensitive (NB4) and ATRA-resistant (NB4-R1) cells. The results showed that reactive oxygen species initiates Ori-induced apoptosis. In addition, we found that neither Ori nor ATRA (10 nM) alone induced marked cell differentiation, while co-treatment of these two compounds can induce differentiation of NB4 and NB4-R1 cells which was accompanied by increased RARalpha, C/EBPepsilon or C/EBPbeta. This is the first report to show that RARalpha could be accumulated by Ori which may be useful as a probe to investigate the mechanism of RARalpha catabolism. These results suggest that Ori is a potential candidate for acute promyelocytic leukemia cancer therapy. PMID- 19302236 TI - Radiographic evaluation of marginal bone levels adjacent to parallel-screw cylinder machined-neck implants and rough-surfaced microthreaded implants using digitized panoramic radiographs. AB - OBJECTIVE: The purpose of this split-mouth study was to compare macro- and microstructure implant surfaces at the marginal bone level during a stress-free healing period and under functional loading. MATERIAL AND METHODS: From January to February 2006, 133 implants (70 rough-surfaced microthreaded implants and 63 machined-neck implants) were inserted in the mandible of 34 patients with Kennedy Class I residual dentitions and followed until February 2008. The marginal bone level was radiographically determined, using digitized panoramic radiographs, at four time points: at implant placement (baseline level), after the healing period, after 6 months of functional loading, and at the end of follow-up. RESULTS: The median follow-up time was 1.9 (range: 1.9-2.1) years. The machined neck group had a mean crestal bone loss of 0.5 mm (range: 0-2.3) after the healing period, 0.8 mm after 6 months (range: 0-2.4), and 1.1 mm (range: 0-3) at the end of follow-up. The rough-surfaced microthreaded implant group had a mean bone loss of 0.1 mm (range: -0.4-2) after the healing period, 0.4 mm (range: 0 2.1) after 6 months, and 0.5 mm (range: 0-2.1) at the end of follow-up. The two implant types showed significant differences in marginal bone levels (healing period: P=0.01; end of follow-up: P<0.01). CONCLUSIONS: Radiographic evaluation of marginal bone levels adjacent to machined-neck or rough-surfaced microthreaded implants showed that implants with the microthreaded design caused minimal changes in crestal bone levels during healing (stress-free) and under functional loading. PMID- 19302237 TI - Degradation of collagen-guided tissue regeneration membranes by proteolytic enzymes of Porphyromonas gingivalis and its inhibition by antibacterial agents. AB - Previous studies have shown that whole cells of several periodontal pathogenic bacteria including Porphyromonas gingivalis may degrade the clinically used regeneration membranes Biomend Extend and Bio-Gide. Fractionation of P. gingivalis cells revealed that cell membrane-associated proteases are responsible for the in vitro degradation of the collagen membranes. In the present study, the specific role of extracellular vesicles and the purified Arg-gingipain enzyme of P. gingivalis in the degradation of three differently cross-linked collagen membranes (Ossix; Bio-Gide and Biomend Extend) was examined. In addition, the inhibitory effect of antibacterial agents and antibiotics used in local periodontal therapy on the enzymatic degradation was evaluated. The data presented show that while all tested collagen membranes, are prone to lysis by oral bacterial proteases, cross-linked membranes are more resistant to proteolysis. Furthermore, therapeutical concentrations of the antibacterial and antibiotic agents chlorhexidine, cetylpyridiniumchloride, minocycline and doxycycline were found to partially inhibit the enzymatic breakdown of the membranes, while metronidazole had no such effect. These results suggest that the presence of P. gingivalis cells, extracellular vesicles and enzymes in the vicinity of regeneration membranes in the periodontium, may change their physical structure and therefore alter their biological properties. Furthermore, the use of cross-linked collagen membranes and antibacterial agents may significantly inhibit this proteolytic process. PMID- 19302238 TI - Instant provisionalization of immediate single-tooth implants is essential to optimize esthetic treatment outcome. AB - OBJECTIVE: The immediate single-tooth implant has become a viable treatment option. However, the impact of the restorative procedure on esthetics is currently unclear. The goal of this study was to compare the soft tissue outline at immediate implants following two restorative protocols: immediate connection of a temporary crown or submerged healing during which a removable partial denture is used. MATERIAL AND METHODS: A 1-year single-blind randomized clinical study was performed in 49 patients. Twenty-four patients were assigned to the immediate restoration group and 25 to the delayed restoration group. Clinical and radiographic evaluations of soft and hard tissues were carried out after 3, 6 and 12 months. RESULTS: Implant survival, bone remodeling, probing depth and bleeding tendency were not influenced by the restorative protocol. Delayed restoration resulted in initial papilla loss taking up to 1 year to attain comparable height as for immediate restoration. Midfacial recession was systematically 2.5-3 times higher following delayed restoration pointing to a 0.75 mm additional loss in comparison with immediate restoration after 1 year. CONCLUSIONS: If the primary implant stability permits it, immediate single-tooth implants should be instantly provisionalized in the interest of optimal midfacial esthetics. PMID- 19302239 TI - Clinical and radiographic comparison of implants in regenerated or native bone: 5 year results. AB - PURPOSE: The aim of this study was to test whether or not implants associated with bone regeneration show the same survival and success rates as implants placed in native bone in patients requiring both forms of therapy. MATERIAL AND METHODS: Thirty-four patients (median age of 60.3 years, range 18-77.7 years) had been treated 5 years before the follow-up examination. Machined screw-type implants were inserted following one of two surgical procedures: (1) simultaneously with a guided bone regeneration (GBR) procedure, which involved grafting with xenogenic bone substitute material, autogenous bone or a mixture of the two and defect covering with a bio-absorbable collagen membrane (test) and (2) standard implantation procedure without bone regeneration (control). For data recording, one test and one control implant from each patient were assessed. Examination included measurements of plaque control record (PCR), probing pocket depth (PPD), bleeding on probing (BOP), width of keratinized mucosa (KM), frequency of situations with supra-mucosal location of the crown margin, implant survival assessment and radiographic examination. Radiographs were digitized to assess the marginal bone level (MBL). Differences between groups were tested using the one-sample t-test. The estimation of survival rate was based on Kaplan Meier analysis. RESULTS: The follow-up period of the 34 GBR and 34 control implants ranged from 49 to 70 months (median time 57 months). Cumulative survival rates reached 100% for the GBR group and 94.1% for the control group without statistical significance. No statistically significant differences for clinical and radiographic parameters were found between the two groups regarding PCR, BOP, PPD, KM and MBL. CONCLUSION: The present study showed that, clinically, implants placed with concomitant bone regeneration did not performed differently from implants placed into native bone with respect to implant survival, marginal bone height and peri-implant soft tissue parameters. PMID- 19302240 TI - Fractalkine functions as a chemoattractant for osteoarthritis synovial fibroblasts and stimulates phosphorylation of mitogen-activated protein kinases and Akt. AB - Fractalkine (FKN/CX3CL1) has been detected in synovial fluids from osteoarthritis (OA) patients. Additionally, low-level expression of the FKN receptor, CX3CR1, has been demonstrated in OA synovial lining. This study aimed to determine a biological function for this ligand/receptor pair in OA and to assess a potential signalling mechanism for FKN in this predominant synovial lining cell type, using chemotaxis assays, Western blotting and F-actin staining. Chemotaxis assays demonstrate that the chemokine domain of FKN effectively induces migration of OA fibroblasts. Consistent with this finding, visualization of F-actin demonstrates that 1 or 10 nM FKN induces noticeable reorganization of cytoskeletal structure in OA fibroblasts after 30 min stimulation with a maximal enhancement at approximately 2 h. In addition, Western blotting analysis demonstrates that FKN stimulates phosphorylation of the mitogen-activated protein (MAP) kinases p38, c Jun N-terminal kinase (JNK) and extracellular-regulated kinase (ERK) 1/2 as well as the serine-threonine kinase Akt at Ser 473 and Thr 308. All these phosphorylation events occur in a time-dependent manner, with little or no activation within 1 min, and maximal activation occurring typically between 5 and 30 min. Moreover, inhibition of ERK 1/2 significantly reduces FKN-induced OA fibroblast migration. These results suggest that FKN is a novel chemoattractant for OA fibroblasts, consistent with FKN-induced alterations in cytoskeletal structure. In addition, FKN induces OA fibroblast signalling via the MAP kinases p38, JNK and ERK 1/2, as well as Akt. PMID- 19302241 TI - Inhibition of indoleamine 2, 3-dioxygenase-mediated tryptophan catabolism accelerates crescentic glomerulonephritis. AB - Immunomodulatory enzyme indoleamine 2, 3-dioxygenase (IDO) is one of the initial and rate-limiting enzymes involved in the catabolism of the essential amino acid tryptophan. Via catalysing tryptophan degradation, IDO suppresses adaptive T cell mediated immunity and plays an important role in various forms of immune tolerance. Its role in T helper type 1 (Th1)-directed, cell-mediated crescentic glomerulonephritis (GN) is still unclear. Therefore, we investigated the activity and role of IDO in crescentic GN using a model of nephrotoxic serum nephritis (NTN), and IDO activity was inhibited by 1-methyl-tryptophan (1-MT) in vivo. Our results showed that activity of IDO, as determined by high performance liquid chromatography analysis of the kynurenine/tryptophan ratio, was increased markedly in the serum and renal tissue of NTN mice, and immunohistochemistry revealed that expression of IDO was up-regulated significantly in glomeruli and renal tubular epithelial cells during NTN. Treatment with 1-MT resulted in significantly exacerbated kidney disease with increased glomerular crescent formation, accumulation of CD4(+)T cells and macrophages in renal tissue, and augmented renal injury compared with phosphate-buffered saline-treated NTN mice, which was associated with enhanced Th1 responses and intrarenal cellular proliferation. These findings suggest that the development of NTN was regulated negatively by increased IDO activity, and IDO might play an important role in the pathogenesis of crescentic GN. PMID- 19302242 TI - Innate immunity and heat shock response in islet transplantation. AB - Islet transplantation is an extremely effective therapy for patients with type I diabetes, providing tight control of blood glucose and persistent insulin release. Islet grafts struggle with various stress responses and immunity attacks, which contribute to loss of islet grafts in the long term. In this review we focus upon the innate immunity and heat shock responses, which are closely relevant to the outcome of islet grafts. Potential strategies provided by more comprehensive interventions to control innate immunity and by selective induction of heat shock proteins may ameliorate the outcome of islet transplantation. PMID- 19302243 TI - Antigenic stimulation with cytochrome P450 2J expressed in mouse hepatocellular carcinoma cells regulates host anti-tumour immunity. AB - Cytochrome P450 2J subfamily (CYP2J) enzymes expressed in mouse hepatocellular carcinoma (HCC) cells were identified as an antigen recognized by specific CD4(+) T cells and the structure of its T cell epitope was determined by proteomics based exploration. The major histocompatibility complex (MHC) class II binding peptides were isolated from I-A(k)/peptide complex of dendritic cells (DCs) loaded or unloaded with MIH-2 mouse HCC cells. MHC class II-binding peptides found in MIH-2-loaded DCs but not in unloaded DCs were determined by tandem mass spectrometric analysis. The peptide, consisting of amino acid 276-290 (DFIDAFLKEMTKYPE) of mouse CYP2J enzymes, was identified as an antigenic peptide presented in the context of MHC class II. Preventive treatment of mice with CYP2J peptide stimulated interferon (IFN)-gamma production of splenocytes and suppressed the growth of implanted CYP2J-positive MIH-2 cells but not CYP2J negative murine bladder tumour cells. However, continuous treatment of MIH-2 bearing mice with CYP2J peptide significantly suppressed IFN-gamma production of splenocytes and accelerated the growth of implanted MIH-2 tumours in vivo. Increased frequencies of CD4(+)forkhead box P3 regulatory T cells and CD11b(+)Gr 1(+) myeloid suppressor cells were observed in splenocytes from the continuously immunized mice. These results indicate that antigenecity of CYP2J isoforms expressed in HCC cells activate host anti-tumour immunity at an initial stage of HCC, but suppress host anti-tumour immunity with excessive antigenic stimulation at an advanced stage. PMID- 19302244 TI - Imbalance between T helper type 17 and T regulatory cells in patients with primary biliary cirrhosis: the serum cytokine profile and peripheral cell population. AB - Primary biliary cirrhosis (PBC) is an organ-specific autoimmune liver disease characterized by progressive loss of intrahepatic small bile ducts. Cellular immune mechanisms involving T cell reaction are thought to be involved significantly in the pathogenesis of PBC. Recent studies have independently revealed enhanced T helper type 17 (Th17) response and weakened T regulatory cell (T(reg)) response in some autoimmune diseases, indicating a role of Th17/T(reg) imbalance in the pathogenesis of autoimmunity. This prompted us to investigate whether the Th17/T(reg) balance was broken in the peripheral blood of patients with PBC and, if it was, what cytokine circumstances might contribute to this imbalance. The expression of 11 Th17/T(reg) differentiation-related genes and serum concentrations of the corresponding cytokines in 36 patients with PBC, 28 patients with chronic hepatitis B and 28 healthy controls were measured by real time quantitative-polymerase chain reaction and enzyme-linked immunosorbent assay respectively. Peripheral Th17 and T(reg) cells were analysed by flow cytometry. Th17-related cytokines were increased significantly in patients with PBC. Consistent with the cytokine profile, the Th17 cell population and retinoid related orphan receptor gammat expression were increased markedly. In contrast, the T(reg) cell population and forkhead box P3 expression were decreased dramatically in the peripheral blood of patients with PBC. Our study revealed that the Th17/T(reg) imbalance, both cytokine profile and cell numbers, exists in patients with PBC, suggesting its potential role in the breakdown of immune self tolerance in PBC. Interleukin-23, which characterized the imbalanced cytokine profile, may play an essential role in Th17-related human autoimmunity. PMID- 19302245 TI - Complement classical pathway components are all important in clearance of apoptotic and secondary necrotic cells. AB - Inherited deficiencies in components of the classical complement pathway are strong disease susceptibility factors for the development of systemic lupus erythematosus (SLE) and there is a hierarchy among deficiency states, the strongest association being with C1q deficiency. We investigated the relative importance of the different complement pathways regarding clearance of apoptotic cells. Phagocytosis of labelled apoptotic Jurkat cells by monocyte-derived macrophages in the presence of sera from individuals with complement deficiencies was studied, as well as C3 deposition on apoptotic cells using flow cytometry. Sera from individuals deficient in C1q, C4, C2 or C3 all showed decreased phagocytosis. Mannose binding lectin (MBL) and the alternative pathway did not influence phagocytosis. Notably, the components of the complement classical pathway, including C1q, were equally important in clearance of apoptotic cells. This indicates that deposition of C3 fragments is of major significance; we therefore studied C3 deposition on apoptotic cells. Experiments with MBL deficient serum depleted of C1q or factor D confirmed the predominance of the classical pathway. At low dilution, sera deficient of C1q, C4 or C2 supported C3 fragment deposition demonstrating alternative pathway activation. In conclusion, we have found that complement-mediated opsonization and phagocytosis of apoptotic cells, particularly those undergoing secondary necrosis, are dependent mainly upon an intact classical pathway. The alternative pathway is less important, but may play a role in some conditions. C1q was not more important than other classical pathway components, suggesting a role in additional pathogenetic processes in SLE other than clearance of apoptotic cells. PMID- 19302246 TI - Innate immunity and inflammation--two facets of the same anti-infectious reaction. AB - Innate immunity is the host's first line of defence against infection. In this review, we present the innate immune response implicated in three examples of pulmonary infection of viral, fungal and bacterial origin. We show that this defence against infection can be a double-edged sword. Thus, the same cells, molecules and mechanisms involved in this protective process can also be involved in deleterious inflammation. A delicate balance between immunity and inflammation is therefore required, making it possible to fight pathogens effectively while limiting inflammation that might be damaging to the host. PMID- 19302247 TI - Genetic and environmental contributions to allergen sensitization in a Chinese twin study. AB - BACKGROUND: Allergic disease is on the rise worldwide. Effective prevention of allergic disease requires comprehensive understanding of the factors that contribute to its intermediate phenotypes, such as sensitization to common allergens. OBJECTIVE: To estimate the degree of genetic and environmental contributions to sensitization to food and aeroallergens. METHODS: Sensitization was defined as a positive skin prick test to an allergen. We calculated the zygosity-specific concordance rates and odds ratios (ORs) for sensitization to food and aeroallergens in 826 Chinese twin pairs [472 monozygotic (MZ) and 354 dizygotic (DZ)] aged 12-28 years. We also applied structural equation modelling procedures to estimate genetic and environmental influences on sensitization. RESULTS: The concordance rates and risk of sensitization in one twin given the presence vs. the absence of sensitization in the other twin were higher in MZ twins than those in DZ twins. However, a large number of MZ twins were discordant in sensitization to common allergens. These observations suggest both genetic and environmental factors influence sensitization. Consistently, the estimated heritability and individual environmental components of the liability to sensitization ranged from 0.51 to 0.68 and 0.32 to 0.49, respectively, based on the best-fitted structural equation model. We also observed high phenotypic correlations between sensitization to two aeroallergens (cockroach and dust mite: 0.83) and two food allergens (peanut and shellfish: 0.58), but only moderate correlations for the pairs between sensitization to a food and an aeroallergen (0.31-0.46). The shared genetic and environmental factors between paired sensitizations contribute to the observed correlations. CONCLUSION: We demonstrated that sensitization to common food and aeroallergens were influenced by both genetic and environmental factors. Moreover, we found that paired allergen sensitizations might share some common sets of genes and environmental factors. This study underscores the need to further delineate unique and/or pleiotropic genetic and environmental factors for allergen sensitization. PMID- 19302248 TI - Soluble human leucocyte antigen-G and interleukin-10 levels in isocyanate-induced asthma. AB - BACKGROUND: We previously reported that in moderate-to-severe asthma there is a deficit of IL-10 secretion that could prevent the production of soluble HLA-G (sHLA-G), a non-classical human leucocyte antigen class I molecule with tissue protective properties in inflammatory responses. OBJECTIVE: Our objective was to investigate the production of sHLA-G and the secretion of IL-10 by peripheral blood mononuclear cells (PBMCs) in asthma induced by isocyanates and to compare the results with those obtained in non-occupational allergic asthma. METHOD: sHLA G and IL-10 were measured by ELISA in the culture supernatants of unstimulated or lipopolysaccharide (LPS)-stimulated PBMCs obtained from 20 subjects with isocyanate asthma, 16 asymptomatic subjects exposed to isocyanates, 18 subjects with non-occupational allergic asthma, and 26 healthy control subjects. RESULTS: Occupational exposure to isocyanates was associated with high baseline levels of secretion of IL-10 by PBMCs, whether or not the exposed subjects had asthmatic symptoms. However, spontaneous production of sHLA-G by PBMC was significantly higher in subjects with isocyanate asthma compared with asymptomatic-exposed controls. In contrast, PBMCs from subjects with non-occupational allergic asthma produced sHLA-G only after LPS stimulation. CONCLUSIONS: sHLA-G production and IL 10 secretion are influenced by workplace exposure to isocyanates and by development of asthma. The different behaviour of both sHLA-G and IL-10 in asthma induced by isocyanates compared with non-occupational allergic asthma suggests a heterogeneous biological role for HLA-G molecules and for IL-10, a key cytokine of immune and inflammatory responses. PMID- 19302249 TI - Safety and tolerability of omalizumab. AB - BACKGROUND: Omalizumab (Xolair) is a recombinant humanized monoclonal anti-IgE antibody with proven efficacy in patients with moderate-to-severe and severe persistent allergic (IgE-mediated) asthma. OBJECTIVE: To review clinical study data to assess the safety profile of omalizumab. METHODS: We analysed the safety of omalizumab using data from completed clinical studies (up to 1 year) involving more than 7500 patients with asthma, rhinitis or related conditions and up to 4 years in one study of patients with severe allergic asthma, as well as post marketing safety data. Analysis focuses on the risk of immune-system effects, hypersensitivity reactions, malignant neoplasia, parasitic infections and thrombocytopenia. RESULTS: Omalizumab exhibited a good safety and tolerability profile that was maintained up to 4 years in one study. The incidence of anaphylaxis was 0.14% in omalizumab-treated patients and 0.07% in control patients. No omalizumab-treated patient developed measurable anti-omalizumab antibodies. Post-marketing, based on estimated exposure of 57,300 patients (June 2003-December 2006), the frequency of anaphylaxis attributed to omalizumab use was estimated to be at least 0.2% of patients. Current clinical trial data do not support an increased risk of malignant neoplasia or thrombocytopenia with omalizumab. CONCLUSION: Data indicate that the proven efficacy of add-on omalizumab in patients with moderate-to-severe or severe allergic asthma is accompanied by a favourable safety and tolerability profile. PMID- 19302250 TI - Evaluation of desloratadine on conjunctival allergen challenge-induced ocular symptoms. AB - BACKGROUND: Allergic conjunctivitis (AC) is frequently associated with allergic rhinitis (AR) and affects up to 91% of patients with AR. Desloratadine, an orally administered, non-sedating second-generation antihistamine, has proven efficacy and safety in the control of nasal and non-nasal--including ocular--symptoms of AR. This randomized-controlled trial is the first to evaluate the effects of continuous oral desloratadine treatment on ocular symptoms after conjunctival allergen challenge. OBJECTIVE: To evaluate the impact of administering oral desloratadine 5 mg daily for 7 days on ocular symptoms in conjunctival allergen challenged subjects. METHODS: In this crossover study, adults with a history of seasonal AC and a minimal threshold response to allergen challenge were randomized to receive desloratadine 5 mg daily (n=20) or placebo (n=21) for 7 days after which they underwent a second ocular allergen challenge. After a 2 week washout period, subjects crossed over to the other treatment. The primary efficacy parameter was the intra-subject difference from baseline at end-point in the post-challenge mean composite ocular redness score (the sum of redness scores in ciliary, conjunctival, or episcleral vessel beds). Secondary efficacy parameters included the intra-subject and inter-subject differences in individual symptom scores for ciliary, conjunctival, or episcleral redness; pruritus; chemosis; eyelid swelling; and tearing. RESULTS: The mean composite ocular redness score was reduced at 10, 15, and 20 min post-challenge with desloratadine treatment compared with placebo, though these differences were not statistically significant (P=NS). Desloratadine was significantly more effective than placebo in preventing post-challenge ocular pruritus (P<0.001). Significant improvements post-challenge were also seen in ciliary and episcleral redness, chemosis, eyelid swelling, and tearing (P<0.05 vs. placebo for all comparisons). Desloratadine was well tolerated. CONCLUSION: Treatment with desloratadine 5 mg daily for 7 days reduced allergic ocular symptoms following allergen challenge. PMID- 19302251 TI - Gene expression studies in cultured dendritic cells: new indicators for the discrimination of skin sensitizers and irritants in vitro. AB - BACKGROUND: The replacement of animal tests for the detection of the sensitizing potential of chemicals is of great importance due to current legislation. One promising approach for the development of an in vitro assay is the exposure of immature dendritic cells (iDCs) to contact sensitizers and irritants, followed by an analysis of the maturation status of the cells. OBJECTIVE: The aim of this study was to further investigate the performance of our previously developed targeted microarray, the immune toxicity chip. In addition, we aimed to identify new marker genes for the discrimination of allergens and irritants using whole genome microarrays. METHODS: Monocyte-derived iDCs were exposed to contact sensitizers and irritants in concentrations resulting in 10-20% cytotoxicity, as determined by dose-response curves. Changes in gene expression were analysed using the immune toxicity chip and a commercially available whole-genome microarray. RESULTS: Using the immune toxicity chip, we could identify a panel of marker genes suitable to discriminate strong allergens and irritants. Analysis with the whole-genome array revealed additional genes that are differentially expressed after allergen exposure, but not after irritant exposure. Hierarchical clustering of these genes showed distinct groups representing the different chemicals. CONCLUSION: Here we show that our test system based on an immune specific microarray is suitable for the discrimination of strong allergens and irritants. Genes detected as differentially expressed with the whole-genome array and previously not connected to the maturation process of DCs might be suitable candidate genes for the identification of weaker sensitizers. PMID- 19302252 TI - Cutaneous manifestations in Hymenoptera and Diptera anaphylaxis: relationship with basal serum tryptase. AB - OBJECTIVES: To compare the clinical presentation of systemic anaphylaxis to Hymenoptera and Diptera with regard to basal serum tryptase (BT) and to evaluate mastocytosis in patients with elevated tryptase. PATIENTS AND METHODS: The medical records of 140 patients with a history of a systemic reaction to venom were retrospectively reviewed. Symptoms and severity of anaphylaxis and BT were recorded. Most patients with elevated tryptase were screened for mastocytosis: a dermatological examination with a skin biopsy was performed in 19 cases and a bone marrow biopsy in 14 cases. RESULTS: Tryptase was elevated in 23 patients. These patients reported fewer usual skin reactions (urticaria in 26.1% of cases with raised tryptase vs. 76.1% of cases with normal tryptase), more flushing (52.2% vs. 4.3%) and frequently did not present skin reaction (26.1% vs. 9.4%). They presented a more severe reaction (mean grade of severity: 3.48 vs. 2.69). Mastocytosis was diagnosed in seven patients with elevated tryptase: indolent systemic mastocytosis in six cases and cutaneous mastocytosis without systemic involvement in one case. In five cases, mastocytosis was previously undiagnosed. Lesions of cutaneous mastocytosis, diagnosed in five patients, consisted of urticaria pigmentosa in all cases and were often inconspicuous. CONCLUSION: These results demonstrate particular clinical features of the allergic reaction in patients with elevated BT and the higher frequency of mastocytosis in this population. In patients with a severe anaphylactic reaction without urticaria, but with flushing, tryptase should be assayed and an underlying mastocytosis should be considered. PMID- 19302253 TI - Beta1-integrins shedding in a guinea-pig model of chronic asthma with remodelled airways. AB - BACKGROUND: A hallmark of airway remodelling in asthma is subepithelial fibrosis, but its relation with airway dysfunction is still controversial. OBJECTIVE: To describe airway functional abnormalities and subepithelial remodelling induced by repetitive antigenic challenges. METHODS: Nine inhaled antigenic challenges were applied every 10 days to guinea-pigs sensitized to ovalbumin (OVA). Antigen induced airway hyperresponsiveness (AI-AHR) to histamine and its immunohistopathological relationship was evaluated at the first, third and ninth OVA challenges. RESULTS: From the first challenge on, OVA induced acute transient bronchoobstruction followed by development of AI-AHR. A progressive rise in baseline Penh (a bronchoobstruction index) and granulocyte airway infiltration was also observed. After the ninth OVA challenge, the amount of extracellular matrix in the subepithelial region (SER) of bronchi and bronchioles was increased. Immunohistochemistry analysis showed that this SER fibrosis was associated to beta1-integrin subunit overexpression, even in acellular areas. Immunoelectronmicroscopy corroborated the location of beta1-integrin in extracellular matrix, essentially in types l and II collagen fibres. Presence of alpha1- and alpha2-integrin subunits in these areas was also corroborated. AI-AHR was correlated with degree of SER increment, cell infiltration, and beta1 integrin expression. CONCLUSION: Our data suggested that beta1-integrin shedding produced by repetitive allergen challenges in guinea-pigs was associated with collagen deposition in SER of bronchi and bronchioles, along with inflammatory cells infiltration and AI-AHR development. PMID- 19302254 TI - Effect of heat treatment and enzymatic digestion on the B cell epitopes of cow's milk proteins. AB - BACKGROUND: Processing milk leads to changes in clinical allergenicity. However, the mechanism by which heat treatment affects the allergenicity of milk proteins is not fully understood. OBJECTIVE: We investigated the effect of heat treatment and enzymatic digestion on the allergenicity of B cell epitopes of milk proteins using a histamine release assay. METHODS: Human basophils were passively sensitized using sera from 10 patients with allergies to cow's milk. All the patients experienced symptoms immediately after ingesting milk. The human basophils were obtained from umbilical cord blood mononuclear cells after culturing the mononuclear cells for 3-4 weeks in the presence of IL-3. After sensitization with 10% patient sera for 48 h, the cells were stimulated with untreated, heat-treated, or heat-treated and pepsin-and-trypsin-digested beta lactoglobulin or alpha-casein for 1 h. The histamine concentrations in the supernatants were then measured by radioimmunoassay. RESULTS: Heat treatment alone did not alter the molecular weight of beta-lactoglobulin or alpha-casein. Heat treatment of beta-lactoglobulin significantly increased its susceptibility to enzymatic digestion in a time- and temperature-dependent manner and reduced its ability to induce histamine release from sensitized basophils. Similar findings were not observed for alpha-casein. The combination of heat treatment and enzymatic digestion reduced the abilities of both beta-lactoglobulin and alpha-casein to induce histamine release from passively sensitized basophils. CONCLUSIONS: Heat treatment reduced the allergenicity of beta-lactoglobulin by inducing conformational changes and by increasing its susceptibility to enzymatic digestion, both of which disrupted B cell epitopes, whereas heat treatment alone did not alter the allergenicity of alpha-casein. PMID- 19302255 TI - High circulating immunoglobulin A levels in infants are associated with intestinal toxigenic Staphylococcus aureus and a lower frequency of eczema. AB - BACKGROUND: Intestinal bacteria trigger IgA production and delayed maturation of mucosal IgA response is linked to allergy development. OBJECTIVE: Our aim was to investigate if plasma levels of IgA or APRIL (a proliferation inducing ligand), an important factor for IgA class switch recombination, in infancy correlates with intestinal colonization by any specific bacteria or yeast. We also examined if plasma IgA or APRIL levels are related to sensitization and the development of eczema. METHODS: IgA was quantified in plasma obtained from infants at birth and at 4 and 18 months of age and APRIL was measured at 4 months of age. Colonization by major bacterial groups and yeast was followed in the first 8 weeks of life by quantitative culture of stool samples. A clinical evaluation regarding the presence of allergen-specific IgE or eczema and eosinophil counts in blood was performed at 18 months of age. RESULTS: In multiple linear regression analysis, only colonization by Staphylococcus aureus strains producing toxins with superantigen function (SEA-D or TSST-1) made an independent contribution to plasma IgA levels at 4 months of age. Further, increased levels of APRIL in plasma at 4 months were negatively associated with sensitization while IgA plasma levels were inversely correlated to eczema development and blood eosinophil counts at 18 months of age. CONCLUSION: Early intestinal colonization by toxigenic S. aureus strains seems to promote systemic IgA responses. Furthermore, high levels of APRIL and IgA in the circulation at 4 months of age seem to correlate negatively with allergy development. PMID- 19302256 TI - Prevalence of allergic rhinitis in 3-6-year-old children in Wuhan of China. AB - BACKGROUND: Only a few prevalence studies of allergic rhinitis (AR) have been reported in China. This study aimed to evaluate the prevalence of AR in a population of 3-6-year-old children in Wuhan, China. METHODS: Sixteen kindergartens in Wuhan City were randomly selected; for each kindergarten, there were three classes from three grades (top, middle and bottom grade, 3-6 years old, respectively). Questionnaires generated by the authors were distributed and filled out by parents of the selected children, with a telephone interview subsequently. Skin prick test (SPT) was carried out on the children after a written consent was signed by the parents. RESULTS: A total of 1211 (89.5%) valid questionnaires were returned for evaluation. The adjusted current prevalence of AR in 3-6-year-old children was 10.8% with the diagnostic criterion of nasal symptoms(+) and SPTs(+). In the SPTs(+) children, the most common inhalant allergen was house dust mites (94.7%), followed by moulds (28.4%). The prevalence of AR was higher in males than that in females (13.0% vs. 7.7%, P < 0.05). 15.8% and 23.2% of AR children were sensitive to egg and milk, respectively. The percentage of children sensitive to both inhalant and food allergens was 27.4%. CONCLUSIONS: We found an unexpectedly high prevalence of diagnosed AR in 3-6-year old children within the investigated population. Dust mite was the most important allergen source for 3-6-year-old children in Wuhan. PMID- 19302257 TI - The low prevalence of allergic disease in Eastern Europe: are risk factors consistent with the hygiene hypothesis? AB - BACKGROUND: The prevalence of allergic disease is known to be low in Eastern Europe. OBJECTIVE: To assess the association of suspected risk factors, including several closely linked to the hygiene hypothesis, with allergic symptoms and atopic sensitization in young school-aged children. METHODS: Observational study of 13 889 Belarusian children followed up at age 6.5 years in the Promotion of Breastfeeding Intervention Trial (PROBIT). Allergic symptoms and diseases were based on parental responses to the International Study of Asthma and Allergy in Childhood questionnaire, and prick tests to five common inhalant allergens were performed using standard methods. RESULTS: Significantly increased risks of wheezing and hayfever symptoms in the past 12 months, and of recurrent itchy rash were observed in boys, children with a positive first-degree family atopic history, and those who had received probiotics (especially as prophylaxis with antibiotic use). Pet ownership, contact with farm animals, the presence and number of younger and (especially) older siblings, and residency in rural areas of Western Belarus were associated with reduced risks. Maternal postnatal smoking was associated with wheezing and hayfever symptoms, while the duration of exclusive breastfeeding was not protective against any of the studied outcomes. The risk factors for allergic symptoms were similar in children with positive skin-prick tests to those in the overall cohort. CONCLUSION: Many of the risk and protective factors we identified are consistent with those reported in Western countries and with the hygiene hypothesis. Further research on dietary and other environmental and genetic factors is necessary to understand the low prevalence of allergic disease in Belarus and other Eastern European countries. PMID- 19302258 TI - Risk factors of adverse reactions to food in German adults. AB - BACKGROUND: Allergic diseases have been increasing during the last decades. Previous studies suggest an impact of a variety of risk factors on the prevalence of food hypersensitivity. OBJECTIVE: Data of a cross-sectional population-based survey were analysed to study the prevalence of food hypersensitivity in females and males adjusted for age and education. METHODS: A population aged 18-79 years from a representative, randomly sampled survey studying 13,300 inhabitants from Germany (Berlin) was analysed. The Berlin study data were weighted with regard to age, sex, education and allergic diseases such as atopic dermatitis, rhinoconjunctivitis and asthma. Instruments for evaluation included mailed questionnaires, structured telephone interviews, physical examinations, detection of total IgE, skin prick tests and double-blind, placebo-controlled food challenge tests (DBPCFC). RESULTS: Three thousand two hundred and twenty-seven questionnaires were evaluated. The data show a significantly higher risk of self reported symptoms in the female group, among persons with higher education and in the age group of 18-39 years. Among individuals with clinical symptoms, females were at lower risk of having positive skin prick tests [aOR=0.32; 95% CI (0.21 0.50); P<0.01] and having a raised total IgE [aOR=0.37; 95% CI (0.24-0.56); P<0.01], but showed a higher risk of non-IgE and IgE-mediated food hypersensitivity [aOR=2.27; 95% CI (1.31-3.93); P<0.01] than males. Based on weighted data, the point prevalence of adverse reactions to food resulted in 3.3% [95% CI (2.4-4.5%)] for women and 1.8% [95% CI (1.2-2.7%)] for men after DBPCFC. CONCLUSION: From a general population survey conducted in Germany, we determined that women are at greater risk of having symptoms of food allergy and also at greater risk of having DBPCFC-confirmed symptomatic food allergy. However, among individuals with symptoms of food allergy, men have a higher prevalence of food specific IgE-sensitization and of raised total IgE than women. PMID- 19302259 TI - Adenosine infusion attenuates soluble RAGE in endotoxin-induced inflammation in human volunteers. AB - AIM: To evaluate possible anti-inflammatory effects of pre-treatment with adenosine in a human experimental inflammatory model. METHODS: The study design was double-blind, crossover, placebo-controlled and randomized. In the Intensive Care Unit of a university hospital, 16 healthy male volunteers were treated for 5.5 h with infusions of adenosine 40 microg kg(-1) min(-1) or placebo. Thirty minutes after the start of adenosine or placebo, 2 ng kg(-1)E-Coli endotoxin was administered. Heart rate, body temperature, blood pressure, plasma cytokines (TNF alpha, IL-6 and IL-10), soluble RAGE and resistin, exhaled nitric oxide and nitrite/nitrate in urine were determined. RESULTS: Endotoxin elicited the expected clinical signs of an inflammatory reaction (tachycardia, fever) and led to prominent release of the cytokines studied (P < 0.001). Resistin in plasma increased after endotoxin (P < 0.001). After placebo treatment, soluble RAGE (sRAGE) in plasma increased 5 h after the endotoxin challenge (P < 0.001) but not after adenosine. After placebo, orally exhaled NO increased with a peak at 4 h (P < 0.001), although there was no statistically significant difference between the two treatments. Nitrite/nitrate in urine (n = 11) did not differ between adenosine and placebo treatments. CONCLUSION: In conclusion, adenosine infusion starting before endotoxin challenge in humans attenuated sRAGE significantly but otherwise had no clear anti-inflammatory effect. Adenosine as a potential anti inflammatory treatment in humans needs further study, including use of higher doses. The mechanism underlying the effect of adenosines on sRAGE remains unknown. PMID- 19302260 TI - Modulation of rabbit sinoatrial node activation sequence by acetylcholine and isoproterenol investigated with optical mapping technique. AB - AIMS: Changes in the rabbit sinoatrial node (SAN) activation sequence with the cholinergic and adrenergic factors were studied. The correlation between the sinus rhythm rate and the leading pacemaker site shift was determined. The hypothesis concerning the cholinergic suppression of nodal cell excitability as one of the mechanisms associated with pacemaker shift was tested. METHODS: A high resolution optical mapping technique was used to register beat-to-beat changes in the SAN activation pattern under the influence of the cholinergic and adrenergic factors. RESULTS: Acetylcholine (10 microm) and strong intramural parasympathetic nerve stimulation caused a pacemaker shift as well as rhythmic slowing and the formation of an inexcitable region in the central part of SAN. In this region the generation of action potentials was suppressed. The slowing of the sinus rhythm (which exceeded 12.8 +/- 3.1% of the rhythm control rate) always accompanied the pacemaker shift. Isoproterenol (10, 100 nm, 1 microm) and sympathetic postganglionic nerve stimulation also evoked a pacemaker shift but without formation of an inexcitable zone. The acceleration of the sinus rhythm, which exceeded 10.5 +/- 1.3% of the control rate of the rhythm, always accompanied the shift. CONCLUSIONS: Both cholinergic and adrenergic factors cause pacemaker shifts in the rabbit SAN. While modest changes in the sinus rhythm do not coincide with the pacemaker shift, greater changes always accompany the shift and may be caused by it, according to one hypothesis. The formation of an inexcitable zone at the place where the leading pacemaker is situated is one of the mechanisms associated with pacemaker shift. PMID- 19302261 TI - No relationship between enzyme activity and structure of nucleotide binding site in sarcoplasmic reticulum Ca(2+)-ATPase from short-term stimulated rat muscle. AB - AIM: We examined whether structural alterations to the adenine nucleotide binding site (ANBS) within sarcoplasmic (endo) reticulum Ca(2+)-ATPase (SERCA) would account for contraction-induced changes in the catalytic activity of the enzyme as assessed in vitro. METHODS: Repetitive contractions were induced in rat gastrocnemius by electrical nerve stimulation. Measurements of sarcoplasmic reticulum properties were performed on control and stimulated muscles immediately after or at 30 min after the cessation of 5-min stimulation. In order to examine the properties at the ANBS, the binding capacity of SERCA to fluorescence isothiocyanate (FITC), a competitive inhibitor at the ANBS, was analysed in microsomes. RESULTS: Short-term electrical stimulation evoked a 23.9% and 32.6% decrease (P < 0.05) in SERCA activity and in the FITC binding capacity, respectively, in the superficial region of the muscle. Whereas SERCA activity reverted to normal levels during 30-min recovery, a restoration of the FITC binding capacity did not occur. CONCLUSION: The discordant changes between the enzyme activity and the FITC binding suggest that, at least during recovery after exercise, changes in SERCA activity may not correlate closely with structural alterations to the ANBS within the enzyme. PMID- 19302262 TI - Small-intestinal bacterial overgrowth in cirrhosis is related to the severity of liver disease. AB - BACKGROUND: Small-intestinal bacterial overgrowth (SIBO) is known to be present in patients with cirrhosis, predisposing to various complications. AIM: To determine the frequency of SIBO in cirrhotics and correlate with severity of cirrhosis. METHODS: Small-intestinal bacterial overgrowth was determined by glucose-hydrogen breath test (GHBT). A basal breath-hydrogen >20 ppm or a rise by > or = 12 ppm above baseline following glucose administration was taken as positive test. Prevalence of SIBO in cirrhotics was compared with healthy controls and correlated with severity of cirrhosis. RESULTS: Of the 53 cirrhotics, 26 (49%) had SIBO, compared to one (8%) control (P = 0.010). The prevalence of SIBO increased with severity of cirrhosis (Child-Pugh A 20%, B 52% and C 73%; P = 0.013). On multivariate analysis, SIBO was independently associated with serum bilirubin and ascites. The best cut-off of serum bilirubin was >/=2 mg/dL [AUROC 0.77 (95% CI 0.64-0.90)] predicting SIBO with sensitivity 65%, specificity 81%, positive predictive value 77%, negative predictive value 71% and accuracy 74%. Patients having combination of ascites and serum bilirubin > or = 2 mg/dL had 82% chance, while patients having neither had only 10% chance of having SIBO. CONCLUSIONS: Small-intestinal bacterial overgrowth was prevalent in about half of cirrhotics. Its frequency increased with increase in severity of cirrhosis. Ascites and raised serum bilirubin reliably predicted presence of SIBO. PMID- 19302263 TI - Systematic review: impaired drug absorption related to the co-administration of antisecretory therapy. AB - BACKGROUND: Due to suppression of gastric acidity during antisecretory therapy, an impaired absorption of co-administered drugs may occur. AIM: To review evidence of impaired drug absorption related to the use of co-administered PPIs or H2RAs. METHODS: Systematic search of MEDLINE/EMBASE/SCOPUS databases (1980 September 2008) for English articles with keywords: drug malabsorption and absorption, stomach, anti-secretory/acid inhibitory drugs, histamine H2 antagonists, PPIs, gastric acid, pH, hypochlorhydria, gastric hypoacidity. From 2126 retrieved articles, 16 randomized crossover studies were identified investigating impaired absorption of nine different drugs in association with co administration of PPIs or H2RAs. Information on investigated drug, study type, features of investigated subjects, study design, type of intervention, and study results were extracted. RESULTS: The identified studies investigated the absorption kinetics of nine drugs. Acid suppression reduced absorption of ketoconazole, itraconazole, atazanavir, cefpodoxime, enoxacin and dipyridamole (median C(max) reduction by 66.5%). An increased absorption of nifedipine and digoxin (median AUC increase by 10%) and a 2-fold-increase in alendronate bioavailability were observed. CONCLUSIONS: Gastric pH appears relevant for absorption of some cardiovascular or infectious disease agents. Antisecretory treatment may significantly modify the absorption of co-administered drugs. PMID- 19302264 TI - Complete recovery of intestinal mucosa occurs very rarely in adult coeliac patients despite adherence to gluten-free diet. AB - BACKGROUND: Expected benefits of gluten-free diet (GFD) in coeliac patients include healing of small intestinal mucosa, but it remains unclear to what extent this benefit is achieved in adults. AIM: To assess factors affecting histological outcome of GFD in a large cohort of adult coeliac patients. METHODS: We extracted information on 465 consecutive coeliac patients studied before and during GFD. RESULTS: Duodenal biopsies at diagnosis were classified as Marsh I in 11, II in 25 and III in 429 cases. After a median 16 months GFD, 38 (8%) patients had histological 'normalization', 300 (65%) had 'remission' with persistent intraepithelial lymphocytosis, 121(26%) had 'no change' and 6 (1%) had 'deterioration'. Coeliac disease related serology was negative in 83% of patients with Marsh III lesion during GFD. Male gender and adherence to GFD were independently associated with histological 'normalization' and 'remission'. Persistence of intraepithelial lymphocytosis was not associated with human lymphocyte antigen gene dose or with Helicobacter pylori infection. CONCLUSIONS: Complete normalization of duodenal lesions is exceptionally rare in adult coeliac patients despite adherence to GFD, symptoms disappearance and negative CD related serology. Control biopsies are mandatory to identify lack of response to gluten free diet. PMID- 19302265 TI - Meta-analysis: octreotide prevents post-ERCP pancreatitis, but only at sufficient doses. AB - BACKGROUND: Effects of octreotide on post-endoscopic retrograde cholangiopancreatography pancreatitis have been studied in many clinical trials. These trials have yielded inconclusive results. Results of more recent studies using larger doses, however, seem to be more optimistic. AIM: To examine effects of octreotide at different doses on PEP. METHODS: A comprehensive search of relevant databases, including Medline, Embase, the Cochrane Controlled Trials Register, the Cochrane Library and Science Citation Index yielded 18 randomized controlled trials (RCTs). Trials were divided into two groups according to the total dosage of octreotide: <0.5 mg (OCT1), > or =0.5 mg (OCT2). The rate of PEP was analysed using a fixed effect model. RESULTS: At doses of > or =0.5 mg, octreotide reduced the rate of PEP. In the OCT2 group, analysis revealed a statistically significant difference on PEP between the octreotide group and the controls (3.4% vs. 7.5%, pooled OR = 0.45; 95% CI: 0.28-0.73; P = 0.001, NNT = 25). In the OCT1 group, the rate of PEP was similar between patients receiving octreotide and the controls (7.2% vs. 6.0%, pooled OR = 1.23; 95% CI: 0.80-1.91; P = 0.35). CONCLUSION: Octreotide is effective in preventing PEP, but only at sufficient doses (> or =0.5 mg). PMID- 19302266 TI - VEGFR1 (Flt1) regulates Rab4 recycling to control fibronectin polymerization and endothelial vessel branching. AB - The cell's main receptor for VEGF, VEGFR2 (Kdr) is one of the most important positive regulators of new blood vessel growth and its downstream signalling is well characterized. By contrast, VEGFR1 (Flt1) and the mechanisms by which this VEGF receptor promotes branching morphogenesis in angiogenesis remain relatively unclear.Here we report that engagement of VEGFR1 activates a Rab4A-dependent pathway that transports alphavbeta3 Integrin from early endosomes to the plasma membrane, and that this is required for VEGF-driven fibronectin polymerization in endothelial cells. Furthermore, VEGFR1 acts to promote endothelial tubule branching in an organotypic model of angiogenesis via a mechanism that requires Rab4A and alphavbeta3 Integrin. We conclude that a recycling pathway regulated by Rab4A is a critical effector of VEGFR1 during branching morphogenesis of the vasculature. PMID- 19302267 TI - Na+/H+ exchangers and RhoA regulate acidic extracellular pH-induced lysosome trafficking in prostate cancer cells. AB - Acidic extracellular pH (pHe) is a common feature of the tumor microenvironment and has been implicated in tumor invasion through the induction of protease secretion.Since lysosomes constitute the major storehouse of cellular proteases, the trafficking of lysosomes to the cell periphery may be required in order to secrete proteases. We demonstrate that a pHe of 6.4-6.8 induced the trafficking of lysosomes to membrane protrusions in the cell periphery. This trafficking event depended upon the PI3K pathway, the GTPase RhoA and sodium-proton exchange activity, resulting in lysosomal exocytosis. Acidic pHe induced a cytoplasmic acidification (although cytoplasmic acidification was not sufficient for acidic pHe-induced lysosome trafficking and exocytosis) and inhibition of NHE activity with the amiloride derivative, EIPA or the anti-diabetic agent troglitazone prevented lysosome trafficking to the cell periphery. Interestingly, using the more specific NHE1 and NHE3 inhibitors, cariporide and s3226 respectively, we show that multiple NHE isoforms are involved in acidic pHe-induced lysosome trafficking and exocytosis. Moreover, in cells expressing NHE1 shRNA, although basal NHE activity was decreased, lysosomes still underwent acidic pHe-induced trafficking,suggesting compensation by other NHE family members.Together these data implicate proton exchangers, especially NHE1 and NHE3, in acidic pHe-induced lysosome trafficking and exocytosis. PMID- 19302268 TI - Requirements for ER-arrest and sequential exit to the golgi of Tomato spotted wilt virus glycoproteins. AB - The envelope glycoproteins Gn and Gc are major determinants in the assembly of Tomato spotted wilt virus (TSWV) particles at the Golgi complex. In this article, the ER-arrest of singly expressed Gc and the transport of both glycoproteins to the Golgi upon coexpression have been analyzed.While preliminary results suggest that the arrest of Gc at the ER (endoplasmic reticulum) did not appear to result from improper folding, transient expression of chimeric Gc, in which the transmembrane domain (TMD) and/or cytoplasmic tail (CT) were swapped for those from Gn, showed that the TMD of Gn was sufficient to allow ER exit and transport to the Golgi. Expression of both glycoproteins in the presence of overexpressed Sar1p specific guanosine nucleotide exchange factor Sec12p, resulted in ER retention demonstrating that the viral glycoproteins are transported to the Golgi in a COPII (coat protein II)-dependent manner. Inhibition of ER Golgi transport by brefeldin A (BFA) had a similar effect on the localization of Gn. However, inhibition of ER (endoplasmic reticulum) to Golgi transport of coexpressed Gc and Gn by overexpression of Sec12p or by BFA revealed distinct localization patterns, i.e. diffuse ER localization versus concentration at specific spots. PMID- 19302269 TI - How location governs toll-like receptor signaling. AB - Toll-like receptors (TLRs) are a family of innate immune system receptors responsible for recognizing conserved pathogen-associated molecular patterns (PAMPs). PAMP binding to TLRs initiates intracellular signaling pathways that lead to the upregulation of a variety of costimulatory molecules and the synthesis and secretion of various cytokines and interferons by cells of the innate immune system. TLR-induced innate immune responses are a prerequisite for the generation of most adaptive immune responses, and in the case of B cells, TLRs directly regulate signaling from the antigen-specific B-cell receptor. The outcome of TLR signaling is determined, in part, by the cells in which they are expressed and by the selective use of signaling adaptors. Recent studies suggest that, in addition, both the ligand recognition by TLRs and the functional outcome of ligand binding are governed by the subcellular location of the TLRs and their signaling adaptors. In this review we describe what is known about the intracellular trafficking and compartmentalization of TLRs in innate system's dendritic cells and macrophages and in adaptive system's B cells, highlighting how location regulates TLR function. PMID- 19302270 TI - Discovery of new cargo proteins that enter cells through clathrin-independent endocytosis. AB - Clathrin-independent endocytosis (CIE) allows internalization of plasma membrane proteins lacking clathrin-targeting sequences, such as the major histocompatibility complex class I protein (MHCI), into cells. After internalization, vesicles containing MHCI fuse with transferrin-containing endosomes generated from clathrin-dependent endocytosis. In HeLa cells, MHCI is subsequently routed to late endosomes or recycled back out to the plasma membrane (PM) in distinctive tubular carriers. Arf6 is associated with endosomal membranes carrying CIE cargo and expression of an active form of Arf6 leads to the generation of vacuolar structures that trap CIE cargo immediately after endocytosis, blocking the convergence with transferrin-containing endosomes. We isolated these trapped vacuolar structures and analyzed their protein composition by mass spectrometry. Here we identify and validate six new endogenous cargo proteins (CD44, CD55, CD98, CD147, Glut1, and ICAM1) that use CIE to enter cells. CD55 and Glut1 appear to closely parallel the trafficking of MHCI, merging with transferrin endosomes before entering the recycling tubules. In contrast, CD44, CD98, and CD147 appear to directly enter the recycling tubules and by-pass the merge with EEA1-positive, transferrin-containing endosomes. This divergent itinerary suggests that sorting may occur along this CIE pathway. Furthermore, the identification of new cargo proteins will assist others studying CIE in different cell types and tissues. PMID- 19302271 TI - Attentional processing of location and color cues during driving. AB - Studies have shown that prior information concerning the spatial location of a subsequent target facilitates the selection of that target for further visual processing in three-dimensional (3-D) space. Using Posner's cuing paradigm, our work examined the reaction time of drivers when cue duration and peripheral environment luminance was changed, and explored the effect color and location on reaction time. Experiment 1 showed that reaction time was not affected by cue duration, and that shifts of attention operated more efficiently for cue location validity. Experiment 2 showed that there was no main effect of background luminance on reaction time. Experiment 3 showed that location produces a greater effect than color. PMID- 19302272 TI - Voriconazole pharmacokinetic variability in cystic fibrosis lung transplant patients. AB - BACKGROUND: Aspergillosis is a high-risk complication in cystic fibrosis (CF) lung transplant patients. Azole antifungal drugs inhibit CYP3A4, resulting in significant metabolic drug-drug interactions. Voriconazole (VRZ) was marketed without therapeutic drug monitoring (TDM) recommendations, consistent with favorable pharmacokinetics, but regular determinations of plasma VRZ concentration were introduced in our center to manage interactions with calcineurin inhibitors and to document the achievement of therapeutic levels. METHODS: VRZ TDM data analysis for trough concentration (C0) and peak concentration (C2) was carried out, using validated liquid chromatography assay with ultraviolet detection, for 35 CF lung transplant patients (mean age 25 years, mean weight 47 kg, balanced sex ratio) since 2003. Therapeutic range (C0: 1.5 +/- 0.5 - C2 : 4.0 +/- 1.0 mg/L) was expressed relative to pivotal pharmacokinetic trial data. RESULTS: The duration of VRZ treatment ranged from 9 days to 22 months. The recommended standard dose of VRZ (200 mg twice a day, following the loading dose) resulted in significant plasma concentrations (>0.5 mg/L) in 20% of CF lung transplant patients. Therapeutic concentrations were obtained using higher doses (average 570 +/- 160 mg/day, +43%, P<0.01). Despite adaptation, C0 remained <0.5 mg/L (11%), even when the drug was administered intravenously, highlighting the variability of VRZ pharmacokinetics, possibly enhanced by CYP2C19 polymorphism. The risk of inefficacy during periods of underdosage was overcome by treatment with antifungal drug combinations (caspofungin, n=10). The therapeutic index was limited by neurologic effects (14%) and hepatic abnormalities (30%). VRZ concentrations correlated significantly (P<0.01) with aspartate aminotransferase levels but not with bilirubin levels. VRZ acted as a metabolic inhibitor of tacrolimus (C0 to dose ratio 5.8 +/- 2.6, n=31/VRZ versus 1.7 +/- 0.9 alone, P<0.001). Large changes in azole concentration affected the magnitude of the drug-drug interactions and adjustment requirements. CONCLUSIONS: TDM is required because VRZ levels are often undetectable in treated CF lung transplant patients, supporting the use of antifungal drug combinations until achievement of VRZ C0 at a steady state between 1 and 2 mg/L. Plasma VRZ concentrations should be determined for the quantitative, individualized management of drug-drug interactions in lung transplant patients, in particular immunosuppressant such as tacrolimus, considering VRZ to be both a target and an inhibitor of CYP3A4. PMID- 19302273 TI - Mycophenolic acid inhibits PMA-induced activation of the neutrophil respiratory burst. AB - Mycophenolate mofetil (MMF) is commonly used in immunosuppressive regimens for solid organ transplantation. There is evidence that the hydrolyzed active agent mycophenolic acid (MPA) causes the endothelial depletion of intracellular guanosine 5'-triphosphate (GTP) levels. This depletion may cause inactivation of nicotinamide adenine dinucleotide phosphate oxidase. The purpose of the present study was to examine the impact of MPA on the neutrophil respiratory burst and phagocytic activity using flow cytometry. In whole blood of healthy volunteers, 2 different doses of MPA (1 and 10 mumol/L) did not alter hydrogen peroxide production of neutrophils induced by receptor-dependent activators. In contrast, MPA inhibits the protein kinase C (PKC)-mediated hydrogen peroxide production by phorbol 12-myristate 13-acetate (PMA) in a time-dependent manner (negative: 21.17 +/- 1.64 vs. 120 min: 14.46 +/- 1.28 mean fluorescence intensity, incubation with 1 mumol/L MPA). In conclusion, our results corroborated that the neutrophil respiratory burst activity of healthy volunteers, induced by either formyl methionyl-leucylphenylalanine (fMLP), priming with tumor necrosis factor alpha followed by fMLP or Escherichia coli and neutrophil phagocytic capacity, were not significantly affected after MPA treatment. We also could demonstrate that the hydrogen peroxide production of neutrophils decreased in response to the PKC activator PMA in a time-dependent manner. PMID- 19302274 TI - A case of disseminated histoplasmosis following autologous stem cell transplantation for Hodgkin's lymphoma: an initial misdiagnosis with a false positive serum galactomannan assay. AB - Systemic histoplasmosis is uncommonly reported in patients who have undergone bone marrow or solid organ transplantation. Diagnosis of systemic histoplasmosis in recipients of transplants may be hampered by lack of consideration of this infection in the differential diagnosis and may be confounded by conflicting information from other testing performed to evaluate for opportunistic infections in this population. We report successful treatment of a case of disseminated histoplasmosis in a patient with Hodgkin's lymphoma who had undergone autologous stem cell transplantation. The diagnosis was delayed by the finding of a positive serum galactomannan assay. PMID- 19302275 TI - The influence of weather conditions on the relative incident rate of fishing vessels. AB - There is a long history of studying the relationship between weather and maritime activities. This article analyzes the link between relative incident rate (RIR) and general weather factors within certain gridded areas and time periods. The study area, which encompasses a broad extent of Atlantic Canadian waters, includes fishing incidents recorded by the Canadian Coast Guard from 1997 to 1999. Methodologies used for traffic track generation in a geographical information system and aggregation of all relevant weather data needed for the statistical analyses are presented. Ultimately, a regression tree was built to illustrate the relationship between incident rate and the following six weather factors: wave height; sea surface temperature; air temperature; ice concentration; fog presence; and precipitation. Results from the regression tree reveal that the RIR defined as (incident number per area-day)/(traffic amount per area-day) across grid cells with incidents, increases as the weather conditions deteriorate in a general way, and the concentration of ice has the biggest influence on the magnitude of incident rates for a given level of traffic exposure. The results from this analysis may assist administrators of maritime traffic, especially those associated with fishing activities, through a better understanding of the influence on RIR of certain weather conditions within given areas in specific time periods. PMID- 19302276 TI - The risk management of third parties during construction in multifunctional urban locations. AB - Buildings above roads, railways, and existing buildings themselves are examples of multifunctional urban locations. The construction stage of those buildings is in general extremely complicated. Safety is one of the critical issues during the construction stage. Because the traffic on the infrastructure must continue during the construction of the building above the infrastructure, falling objects due to construction activities form a major hazard for third parties, i.e., people present on the infrastructure or beneath it, such as car drivers and passengers. This article outlines a systematic approach to conduct quantitative risk assessment (QRA) and risk management of falling elements for third parties during the construction stage of the building above the infrastructure in multifunctional urban locations. In order to set up a QRA model, quantifiable aspects influencing the risk for third parties were determined. Subsequently, the conditional probabilities of these aspects were estimated by historical data or engineering judgment. This was followed by integrating those conditional probabilities, now used as input parameters for the QRA, into a Bayesian network representing the relation and the conditional dependence between the quantified aspects. The outcome of the Bayesian network-the calculation of both the human and financial risk in quantitative terms-is compared with the risk acceptance criteria as far as possible. Furthermore, the effect of some safety measures were analyzed and optimized in relation with decision making. Finally, the possibility of integration of safety measures in the functional and structural building design above the infrastructure are explored. PMID- 19302277 TI - Policy learning for flood mitigation: a longitudinal assessment of the community rating system in Florida. AB - Floods continue to inflict the most damage upon human communities among all natural hazards in the United States. Because localized flooding tends to be spatially repetitive over time, local decisionmakers often have an opportunity to learn from previous events and make proactive policy adjustments to reduce the adverse effects of a subsequent storm. Despite the importance of understanding the degree to which local jurisdictions learn from flood risks and under what circumstances, little if any empirical, longitudinal research has been conducted along these lines. This article addresses the research gap by examining the change in local flood mitigation policies in Florida from 1999 to 2005. We track 18 different mitigation activities organized into four series of activities under the Federal Emergency Management Agency's (FEMA) Community Rating System (CRS) for every local jurisdiction in Florida participating in the FEMA program on a yearly time step. We then identify the major factors contributing to policy changes based on CRS scores over the seven-year study period. Using multivariate statistical models to analyze both natural and social science data, we isolate the effects of several variables categorized into the following groups: hydrologic conditions, flood disaster history, socioeconomic and human capital controls. Results indicate that local jurisdictions do in fact learn from histories of flood risk and this process is expedited under specific conditions. PMID- 19302278 TI - Managing societal and institutional risk in child protection. AB - Public sector services have been reshaped by two interacting factors: the growing dominance of risk management and the growing demands for transparency and accountability. For the caring professions, these have provoked radical reform. Using the child protection service as a case study, this article explores the impact of the changes on a service that deals with conflicting risks and has a poorly articulated knowledge base. Drawing on Rothstein et al.'s distinction between societal and institutional risks, it is argued that difficulties in managing societal risks are creating serious institutional risks. The latter are then being prioritized in the way the system operates. The preoccupation with such risks has been translated into concerted efforts to formalize the work of front line practitioners to make it transparent and auditable. Although done, in part, with the good intention of spreading good practice standards, this formalization has gone beyond the evidenced knowledge base to the extent that it is creating a new picture of "good practice" that omits significant dimensions of work and is distinct from measures of children's safety or welfare. Moreover, the process of formalization acts as an impediment to knowledge development in disciplines where such learning is urgently needed. PMID- 19302279 TI - Uncertainty analysis based on probability bounds (p-box) approach in probabilistic safety assessment. AB - A wide range of uncertainties will be introduced inevitably during the process of performing a safety assessment of engineering systems. The impact of all these uncertainties must be addressed if the analysis is to serve as a tool in the decision-making process. Uncertainties present in the components (input parameters of model or basic events) of model output are propagated to quantify its impact in the final results. There are several methods available in the literature, namely, method of moments, discrete probability analysis, Monte Carlo simulation, fuzzy arithmetic, and Dempster-Shafer theory. All the methods are different in terms of characterizing at the component level and also in propagating to the system level. All these methods have different desirable and undesirable features, making them more or less useful in different situations. In the probabilistic framework, which is most widely used, probability distribution is used to characterize uncertainty. However, in situations in which one cannot specify (1) parameter values for input distributions, (2) precise probability distributions (shape), and (3) dependencies between input parameters, these methods have limitations and are found to be not effective. In order to address some of these limitations, the article presents uncertainty analysis in the context of level-1 probabilistic safety assessment (PSA) based on a probability bounds (PB) approach. PB analysis combines probability theory and interval arithmetic to produce probability boxes (p-boxes), structures that allow the comprehensive propagation through calculation in a rigorous way. A practical case study is also carried out with the developed code based on the PB approach and compared with the two-phase Monte Carlo simulation results. PMID- 19302280 TI - The association of knowledge with concern about global warming: trusted information sources shape public thinking. AB - During the last decade, a great deal of news media attention has focused on informing the American public about scientific findings on global warming (GW). Has learning this sort of information led the American public to become more concerned about GW? Using data from two surveys of nationally representative samples of American adults, this article shows that the relation between self reported knowledge and concern about GW is more complex than what previous research has suggested. Among people who trust scientists to provide reliable information about the environment and among Democrats and Independents, increased knowledge has been associated with increased concern. But among people who are skeptical about scientists and among Republicans more knowledge was generally not associated with greater concern. The association of knowledge with concern among Democrats and Independents who trust scientists was mediated by perceptions of consensus among scientists about GW's existence and by perceptions that humans are a principal cause of GW. Moreover, additional analyses of panel survey data produced findings consistent with the notion that more knowledge yields more concern among Democrats and Independents, but not among Republicans. Thus, when studying the relation of knowledge and concern, it is important to take into account the content of the information that different types of people acquire and choose to rely upon. PMID- 19302281 TI - Detection and treatment of Strongyloides hyperinfection syndrome following lung transplantation. AB - Strongyloides hyperinfection syndrome has not been reported in lung transplant recipients. We describe the case of a 61-year-old Peruvian man, who received bilateral lung transplants for idiopathic pulmonary fibrosis, and subsequently developed persistent fever with pulmonary infiltrates, ventilator dependence, and pneumothoraces. Bronchoalveolar lavage (BAL) cultures for bacteria, viruses, and fungi were negative, but testing for ova and parasites from BAL fluid revealed Strongyloides stercoralis larvae on day 16 post transplant. He was successfully treated with albendazole and ivermectin, and immunosuppression was reduced. BAL fluid also grew Mycobacterium kansasii, for which he received combination anti mycobacterial therapy. This case illustrates the importance of screening for parasitic infections before transplantation in the appropriate clinical setting, and demonstrates the utility of direct diagnostic evaluation for parasitic infections in at-risk post-transplant patients with unexplained illnesses. PMID- 19302282 TI - Pseudomonas aeruginosa bacteremia over a 10-year period: multidrug resistance and outcomes in transplant recipients. AB - AIM: Transplant recipients are at risk for hospital-acquired infections (HAIs), including those caused by Pseudomonas aeruginosa. Of all HAIs, bloodstream infection (BSI) remains one of the most life-threatening. METHODS: Over a 10-year period, we studied 503 patients, including 149 transplant recipients, with pseudomonal BSI from the University of Pittsburgh Medical Center. Trends in antimicrobial susceptibility, risk factors for multidrug resistance (MDR), and outcomes were compared between transplant and non-transplant patients. RESULTS: Resistance to all antibiotic classes was significantly greater in pseudomonal blood culture isolates from transplant compared with non-transplant patients (P<0.001). Of isolates from transplant recipients (n=207), 43% were MDR, compared with 18% of isolates from non-transplant patients (n=391) (odds ratio [OR] 3.47; 95% confidence interval [CI] 2.34-5.14, P<0.001). Among all patients, independent risk factors for MDR P. aeruginosa BSI included previous transplantation (OR 2.38; 95% CI 1.51-3.76, P<0.001), hospital-acquired BSI (OR 2.41; 95% CI 1.39 4.18, P=0.002), and prior intensive care unit (ICU) admission (OR 2.04; 95% CI 1.15-3.63, P=0.015). Mortality among transplant recipients was 42%, compared with 32% in non-transplant patients (OR 1.55; 95% CI 0.87-2.76, P=0.108). For transplant recipients, onset of BSI in the ICU was the only independent predictor of mortality (OR 8.00; 95% CI 1.71-37.42, P=0.008). CONCLUSIONS: Transplant recipients are at greater risk of MDR P. aeruginosa BSI, with an appreciable mortality. Future management must concentrate on the implementation of effective preventative strategies. PMID- 19302284 TI - Cellular senescence: its role in tumor suppression and aging. AB - In normal tissue, cell division is carefully regulated to maintain the correct proliferative balance. Abnormal cell division underlies many hypoproliferative and hyperproliferative disorders, including cancer, and a better understanding of the mechanisms involved could lead to new strategies for treatment and prevention. Cellular senescence, a state of irreversible growth arrest, was first described as a limit to the replicative life span of somatic cells after serial cultivation in vitro. Recently, however, it has also been shown to be triggered prematurely by potentially oncogenic stimuli such as oncogene expression, oxidative stress, and DNA damage in cell culture studies. These data suggest that cellular senescence is therefore acting as a tumor-protective fail-safe mechanism. However, the significance of cellular senescence has remained an issue of debate over the years, with the possibility that it might be a cell culture related artifact. Recent reports on oncogene-induced senescence detected in premalignant tumors have provided evidence to validate its role as a physiological response to prevent oncogenesis in vivo. In this review, we discuss the mechanisms for cellular senescence and its roles in vivo. PMID- 19302283 TI - Multicolor in vivo targeted imaging to guide real-time surgery of HER2-positive micrometastases in a two-tumor coincident model of ovarian cancer. AB - One of the primary goals of oncological molecular imaging is to accurately identify and characterize malignant tissues in vivo. Currently, molecular imaging relies on targeting a single molecule that while overexpressed in malignancy, is often also expressed at lower levels in normal tissue, resulting in reduced tumor to background ratios. One approach to increasing the specificity of molecular imaging in cancer is to use multiple probes each with distinct fluorescence to target several surface antigens simultaneously, in order to identify tissue expression profiles, rather than relying on the expression of a single target. This next step forward in molecular imaging will rely on characterization of tissue based on fluorescence and therefore will require the ability to simultaneously identify several optical probes each attached to different targeting ligands. We created a novel 'coincident' ovarian cancer mouse model by coinjecting each animal with two distinct cell lines, HER2+/red fluorescent protein (RFP)- SKOV3 and HER2-/RFP+ SHIN3-RFP, in order to establish a model of disease in which animals simultaneously bore tumors with two distinct phenotypes (HER2+/RFP-, HER2-/RFP+), which could be utilized for multicolor imaging. The HER2 receptor of the SKOV3 cell line was targeted with a trastuzumab-rhodamine green conjugate to create green tumor implants, whereas the RFP plasmid of the SHIN3 cells created red tumor implants. We demonstrate that real-time in vivo multicolor imaging is feasible and that fluorescence characteristics can then serve to guide the surgical removal of disease. PMID- 19302285 TI - Chemotherapeutic agents potentiate adenoviral gene therapy for pancreatic cancer. AB - Adenovirus-mediated gene therapy combined with chemotherapeutic agents is expected to represent a new approach for treating pancreatic cancer. However, there have been no reports of definitive effects of chemotherapeutic agents on adenovirus-mediated gene therapies. In the present study, we investigated the effects of chemotherapeutic agents on the transduction efficiency of an adenovirus-based gene therapy. Adenovirus (Ad-NK4) expressing NK4, which acts as a hepatocyte growth factor antagonist, was used as a representative gene therapy. Pancreatic cancer cells infected with Ad-NK4 were treated with chemotherapeutic agents (5-fluorouracil [5FU], cisplatin or etoposide), and the NK4 levels in their culture media were measured. To examine the effects of chemotherapeutic agents in vivo, Ad-NK4 was administered to subcutaneous tumors in mice after treatment with the agents, and the tumor NK4 levels were measured. The NK4 levels in culture media from cells treated with 5FU, cisplatin and etoposide were 5.2 fold (P = 0.026), 6-fold (P < 0.001) and 4.3-fold (P < 0.001) higher than those of untreated cells, respectively. The chemotherapeutic agents also increased Ad NK4 uptake. The NK4 levels in tumors treated with 5FU, cisplatin and etoposide were 5.4-fold (P = 0.006), 11.8-fold (P < 0.001) and 4.9-fold (P = 0.017) higher than those in untreated tumors, respectively. The present findings suggest that chemotherapeutic agents significantly improve the efficiency of adenovirus mediated gene transfer in pancreatic cancer. Furthermore, they will contribute to decreases in the adenovirus doses required for gene transfer, thereby controlling the side-effects of adenovirus infection in normal tissues. PMID- 19302286 TI - Leukemia-related transcription factor TEL/ETV6 expands erythroid precursors and stimulates hemoglobin synthesis. AB - TEL/ETV6 located at chromosome 12p13 encodes a member of the E26 transformation specific family of transcription factors. TEL is known to be rearranged in a variety of leukemias and solid tumors resulting in the formation of oncogenic chimeric protein. Tel is essential for maintaining hematopoietic stem cells in the bone marrow. To understand the role of TEL in erythropoiesis, we generated transgenic mice expressing human TEL under the control of Gata1 promoter that is activated during the course of the erythroid-lineage differentiation (GATA1-TEL transgenic mice). Although GATA1-TEL transgenic mice appeared healthy up to 18 months of age, the level of hemoglobin was higher in transgenic mice compared to non-transgenic littermates. In addition, CD71+/TER119+ and c-kit+/CD41+ populations proliferated with a higher frequency in transgenic mice when bone marrow cells were cultured in the presence of erythropoietin and thrombopoietin, respectively. In transgenic mice, enhanced expression of Alas-e and beta-major globin genes was observed in erythroid-committed cells. When embryonic stem cells expressing human TEL under the same Gata1 promoter were differentiated into hematopoietic cells, immature erythroid precursor increased better compared to controls as judged from the numbers of burst-forming unit of erythrocytes. Our findings suggest some roles of TEL in expanding erythroid precursors and accumulating hemoglobin. PMID- 19302287 TI - Methylation of estrogen receptor 1 in colorectal adenomas is not age-dependent, but is correlated with K-ras mutation. AB - The promoter region of estrogen receptor 1 (ESR1) has been shown to be methylated in normal colorectal mucosa in an age-dependent manner. However, the methylation of this region in colorectal tumors has not sufficiently been investigated. The methylation status of ESR1 in 105 colorectal adenoma tissues was examined by MethyLight and presented as the percentage of methylated references (PMR). Factors that affect the PMR of ESR1 in adenomas were determined using parameters including patient age, sex, past history of malignancy, family history of colorectal cancer, smoking and drinking habits, clinical characteristics of adenomas (location, size, macroscopic appearance, and histology), and K-ras mutation. Multiple linear regression revealed that the PMR was not correlated with patient age. K-ras mutation was significantly correlated with the higher methylation status of ESR1 in adenoma (t-value = 3.21, P = 0.0018), whereas alcohol exposure was significantly correlated with lower methylation status (t value = -2.37, P = 0.02). Because methylation of O6-methylguanine DNA methyltransferase (MGMT) has been reported to be correlated with K-ras G-to-A transition, methylation of ESR1 was compared with that of MGMT with regard to K ras mutation. Contrary to expectations, methylation of MGMT was not significantly correlated with K-ras G-to-A transition, but that of ESR1 was strongly correlated with K-ras G-to-A transition. Thus, the methylation status of ESR1 in adenomas was not correlated with patient age, but was associated with K-ras mutation, suggesting that methylation of ESR1 in tumors functions differently from that in normal colon mucosa. PMID- 19302288 TI - Combination therapy of in vitro-expanded natural killer T cells and alpha galactosylceramide-pulsed antigen-presenting cells in patients with recurrent head and neck carcinoma. AB - The aim of this clinical trial was to investigate the feasibility of intra arterial infusion of in vitro-expanded Valpha24 natural killer T (NKT) cells combined with submucosal injection of alpha-galactosylceramide (KRN7000; alphaGalCer)-pulsed antigen-presenting cells (APC). A phase I clinical study was carried out in patients with head and neck squamous cell carcinoma (HNSCC). Patients with locally recurrent HNSCC refractory to standard therapy were eligible. Eight patients received super-selective transcatheter intra-arterial infusion of activated Valpha24 NKT cells into tumor-feeding arteries and nasal submucosal injections of alphaGalCer-pulsed APC twice with a 1-week interval. Valpha24 NKT cell-specific immune responses, safety, and antitumor effects were evaluated. The number of Valpha24 NKT cells and interferon-gamma-producing cells in peripheral blood mononuclear cells increased in seven out of eight patients enrolled. Grade 3 toxicity with a pharyngocutaneous fistula related to local tumor reduction was observed in one patient and mild adverse events with grade 1 2 symptoms occurred in seven patients. Regarding the clinical responses, three cases exhibited a partial but significant response, four were classified as stable disease, and one patient continued to develop progressive disease. The use of the intra-arterial infusion of activated Valpha24 NKT cells and the submucosal injection of alphaGalCer-pulsed APC has been shown to induce significant antitumor immunity and had beneficial clinical effects in the management of advanced HNSCC. The use of such therapeutic modalities may be helpful in the management of tumors and therefore needs to be explored in further detail. The clinical trial registration number was UMIN000000722. PMID- 19302289 TI - Platelets and fibrinogen facilitate each other in protecting tumor cells from natural killer cytotoxicity. AB - The functions of platelets and fibrinogen in protecting tumor cells from natural killer cytotoxicity have been discussed for more than 20 years. However, their exact roles and relationships in the process are still not clear. In this study, we show that tumor cells prefer to adhere to fibrinogen than to platelets, and fibrinogen can enhance the adhesion of tumor cells to platelets. Beta3 integrin plays an important role in the adhesion of B16F10 to platelets enhanced by fibrinogen. In the presence of thrombin, fibrinogen forms dense fibrin(ogen) layers around tumor cells. Tumor cells can induce platelets to aggregate and form thrombin. Platelets, as well as thrombin, can help fibrinogen protect tumor cells from lethal contact with natural killer cells and natural killer cytotoxicity. Hirudin, a specific inhibitor of thrombin, can reverse the effect of platelets on fibrinogen in blocking natural killer cytotoxicity. Our results suggest that fibrinogen helps platelets to adhere to tumor cells, and platelets in turn promote more fibrinogen to aggregate around tumor cells by forming thrombin. They facilitate each other in protecting tumor cells from natural killer cytotoxicity. PMID- 19302290 TI - Core fucosylation of E-cadherin enhances cell-cell adhesion in human colon carcinoma WiDr cells. AB - Alpha1,6-fucosyltransferase (Fut8), an enzyme that catalyzes the introduction of alpha1,6 core fucose to the innermost N-acetylglucosamine residue of the N glycan, has been implicated in the development, immune system, and tumorigenesis. We found that alpha1,6-fucosyltransferase and E-cadherin expression levels are significantly elevated in primary colorectal cancer samples. Interestingly, low molecular weight population of E-cadherin appeared as well as normal sized E cadherin in cancer samples. To investigate the correlation between alpha1,6 fucosyltransferase and E-cadherin expression, we introduced alpha1,6 fucosyltransferase in WiDr human colon carcinoma cells. It was revealed that the low molecular weight population of E-cadherin was significantly increased in alpha1,6-fucosyltransferase-transfected WiDr cells in dense culture, which resulted in an enhancement in cell-cell adhesion. The transfection of mutated alpha1,6-fucosyltransferase with no enzymatic activity had no effect on E cadherin expression, indicating that core fucosylation is involved in the phenomena. In alpha1,6-fucosyltransferase knock down mouse pancreatic acinar cell carcinoma TGP49 cells, the expression of E-cadherin and E-cadherin dependent cell cell adhesion was decreased. The introduction of alpha1,6-fucosyltransferase into kidney epithelial cells from alpha1,6-fucosyltransferase(-/-) mice restored the expression of E-cadherin and E-cadherin-dependent cell-cell adhesion. Based on the results of lectin blotting, peptide N-glycosidase F treatment, and pulse chase studies, it was demonstrated that the low molecular weight population of E cadherin contains peptide N-glycosidase F insensitive sugar chains, and the turnover rate of E-cadherin was reduced in alpha1,6-Fucosyltransferase transfectants. Thus, it was suggested that core fucosylation regulates the processing of oligosaccharides and turnover of E-cadherin. These results suggest a possible role of core fucosylation in the regulation of cell-cell adhesion in cancer. PMID- 19302291 TI - Down-regulation of cIAP2 enhances 5-FU sensitivity through the apoptotic pathway in human colon cancer cells. AB - Currently 5-fluorouracil (5-FU) plays a central role in the chemotherapeutic regimens for colorectal cancers and thus it is important to understand the mechanisms that determine 5-FU sensitivity. The expression profiles of human colon cancer cell line DLD-1, its 5-FU-resistant subclone DLD-1/FU and a further 21 types of colon cancer cell lines were compared to identify the novel genes defining the sensitivity to 5-FU and to estimate which population of genes is responsible for 5-FU sensitivity. In the hierarchical clustering, DLD-1 and DLD 1/FU were most closely clustered despite over 100 times difference in their 50% inhibitory concentration of 5-FU. In DLD-1/FU, the population of genes differentially expressed compared to DLD-1 was limited to 3.3%, although it ranged from 4.8% to 24.0% in the other 21 cell lines, thus indicating that the difference of 5-FU sensitivity was defined by a limited number of genes. Next, the role of the cellular inhibitor of apoptosis 2 (cIAP2) gene, which was up regulated in DLD-1/FU, was investigated for 5-FU resistance using RNA interference. The down-regulation of cIAP2 efficiently enhanced 5-FU sensitivity, the activation of caspase 3/7 and apoptosis under exposure to 5-FU. The immunohistochemistry of cIAP2 in cancer and corresponding normal tissues from colorectal cancer patients in stage III revealed that cIAP2 was more frequently expressed in cancer tissues than in normal tissues, and cIAP2-positive patients had a trend toward early recurrence after fluorouracil-based chemotherapy. Although the association between drug sensitivity and the IAP family in colorectal cancer has not yet been discussed, cIAP2 may therefore play an important role as a target therapy in colorectal cancer. PMID- 19302292 TI - Pancreatic adenocarcinoma up-regulated factor (PAUF), a novel up-regulated secretory protein in pancreatic ductal adenocarcinoma. AB - The identification of novel tumor-specific proteins or antigens is of great importance for diagnostic and therapeutic applications in pancreatic cancer. Using oligonucleotide microarrays, we identified a broad spectrum of differentially expressed pancreatic cancer-related genes. Of these, we selected an overexpressed expressed sequence taq and cloned a 721-bp full-length cDNA with an open reading frame of 196 amino acids. This novel gene was localized on the Homo sapiens 16p13.3 chromosomal locus, and its nucleotide sequence matched the Homo sapiens similar to common salivary protein 1 (LOC124220). We named the gene pancreatic adenocarcinoma up-regulated factor. The pancreatic adenocarcinoma up regulated factor was secreted into the culture medium of pancreatic adenocarcinoma up-regulated factor-overexpressing Chinese hamster ovary cells, had an apparent molecular mass of approximately 25 kDa, and was N-glycosylated. The induction of pancreatic adenocarcinoma up-regulated factor in Chinese hamster ovary cells increased cell proliferation, migration, and invasion ability in vitro. Subcutaneous injection of mice with Chinese hamster ovary/pancreatic adenocarcinoma up-regulated factor cells resulted in 3.8-fold greater tumor sizes compared to Chinese hamster ovary/mock cells. Reverse transcription-polymerase chain reaction and western blotting with antirecombinant human pancreatic adenocarcinoma up-regulated factor antibodies confirmed that pancreatic adenocarcinoma up-regulated factor was highly expressed in six of eight pancreatic cancer cell lines. Immunohistochemical staining of human pancreatic cancer tissues also showed pancreatic adenocarcinoma up-regulated factor overexpression in the cytoplasm of cancer cells. Transfection with pancreatic adenocarcinoma up-regulated factor-specific small-interfering RNA reduced cancer cell migration and invasion in vitro. Treatment with antirecombinant human pancreatic adenocarcinoma up-regulated factor in vitro and in vivo reduced proliferation, migration, invasion, and tumorigenic ability. Collectively, our results suggest that pancreatic adenocarcinoma up-regulated factor is a novel secretory protein involved in pancreatic cancer progression and might be a potential target for the treatment of pancreatic cancer. PMID- 19302293 TI - Isolation and sources of 'blown pack' spoilage clostridia in beef abattoirs. AB - AIMS: (i) To evaluate methods for isolation and molecular detection of blown pack spoilage (BPS) clostridia and (ii) to survey beef abattoirs for sources and distributions of Clostridium estertheticum and Cl. gasigenes. METHODS AND RESULTS: Molecular detection and conventional isolation methods were used to detect and recover BPS associated clostridia (Cl. estertheticum and Cl. gasigenes), from four commercial Irish beef abattoirs and their environments, during a one year study. DNA-based methods detected 218 Cl. estertheticum and 300 Cl. gasigenes, from 1680 samples, whereas culture-methods only yielded 17 Cl. estertheticum and 176 Cl. gasigenes isolates. BPS Clostridia were frequently detected in beef abattoirs and their environments, especially at areas prior to hide removal. The study noted a higher percentage of positive samples during the month of May (38.6%). CONCLUSIONS: (i) DNA-based techniques are the most reliable ways to determine the presence of these organisms in various samples and (ii) hides and faeces are the main reservoirs of BPS clostridia in the abattoirs. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper provides useful information to detect BPS organisms, as well as to develop a science-based control strategy of the problem. PMID- 19302294 TI - Evaluation of hydrogen peroxide-based disinfectants in a new resazurin microplate method for rapid efficacy testing of biocides. AB - AIMS: Development of the resazurin microplate method (RMM) as a novel test system for the evaluation of the antimicrobial activity of antiseptics and disinfectants. The validated RMM was subsequently applied for the evaluation of hydrogen peroxide (H(2)O(2)) and stabilized H(2)O(2) combination products. METHODS AND RESULTS: The European Committee for Standardization prescribes the plate count challenge test (PCCT) for antiseptic and disinfectant efficacy testing. This protocol was adapted to a microplate-based assay, using resazurin as viability indicator. The RMM was as accurate as the PCCT, had an identical detection limit and showed high intermediate precision. Using the validated RMM, it was shown that H(2)O(2) combined with silver possessed a higher bactericidal and fungicidal activity compared to native H(2)O(2) with and without glycerol. CONCLUSIONS: Validation showed that the RMM may replace the PCCT. When applying the RMM, H(2)O(2) combined with silver was clearly a more potent disinfectant compared to H(2)O(2) in killing bacteria and fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: The RMM is easier to use for antimicrobial efficacy testing of antiseptics and disinfectants. As the RMM is in accordance with the norms of the European Committee for Standardization, it may become a more cost-effective alternative to the more laborious PCCT reference method. H(2)O(2) with silver may replace native H(2)O(2) to increase overall disinfection efficiency. PMID- 19302295 TI - Influence of different carbon sources on bacterial cellulose production by Gluconacetobacter xylinus strain ATCC 53524. AB - AIMS: To determine the effect of carbon sources on cellulose produced by Gluconacetobacter xylinus strain ATCC 53524, and to characterize the purity and structural features of the cellulose produced. METHODS AND RESULTS: Modified Hestrin Schramm medium containing the carbon sources mannitol, glucose, glycerol, fructose, sucrose or galactose were inoculated with Ga. xylinus strain ATCC 53524. Plate counts indicated that all carbon sources supported growth of the strain. Sucrose and glycerol gave the highest cellulose yields of 3.83 and 3.75 g l(-1) respectively after 96 h fermentation, primarily due to a surge in cellulose production in the last 12 h. Mannitol, fructose or glucose resulted in consistent rates of cellulose production and yields of >2.5 g l(-1). Solid state (13)C CP/MAS NMR revealed that irrespective of the carbon source, the cellulose produced by ATCC 53524 was pure and highly crystalline. Scanning electron micrographs illustrated the densely packed network of cellulose fibres within the pellicles and that the different carbon sources did not markedly alter the micro architecture of the resulting cellulose pellicles. CONCLUSIONS: The production rate of bacterial cellulose by Ga. xylinus (ATCC 53524) was influenced by different carbon sources, but the product formed was indistinguishable in molecular and microscopic features. SIGNIFICANCE AND IMPACT OF THE STUDY: Our studies for the first time examined the influence of different carbon sources on the rate of cellulose production by Ga. xylinus ATCC 53524, and the molecular and microscopic features of the cellulose produced. PMID- 19302296 TI - Biochemical and genetic variation in Mycoplasma fermentans strains from cell line, human and animal sources. AB - AIMS: To investigate the inter-strain variation in (i) substrate utilization and (ii) the restriction fragment length polymorphism (RFLP) pattern based on the distribution of an insertion element (IS1550) in Mycoplasma fermentans strains, and to establish any correlation between subgroups within the species and their source or habitat. METHODS AND RESULTS: Using a sensitive dynamic pH method, the pattern and kinetics of substrate utilization by a panel of 17 M. fermentans strains from various sources was determined. This study correlated the biochemical characteristics of these strains with RFLP patterns based on the distribution of an insertion sequence (IS1550) with the sources of the strains. The test isolates were divided into four major groups according to the pattern of substrates metabolized. Interestingly, two strains isolated from cell lines in RFLP cluster I failed to utilize arginine. Ovine strains showed distinct substrate utilization patterns and produced RFLP patterns not previously encountered. CONCLUSIONS: All strains utilized glucose, but the ability to utilize arginine, fructose and N-acetyl glucosamine varied. There was also some correlation evident between the metabolic data and the RFLP clusters. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has provided a better understanding of the biochemical and genetic diversity of M. fermentans strains from various sources. PMID- 19302297 TI - Induction of the histidine decarboxylase genes of Photobacterium damselae subsp. damselae (formally P. histaminum) at low pH. AB - AIMS: To elucidate the detailed mechanism of histamine production by Photobacterium damselae subsp. damselae. METHODS AND RESULTS: Histidine decarboxylase and related genes of P. damselae subsp. damselae were cloned, and three open reading frames named as hdcT, hdcA and hisRS were identified. The hdcA gene encodes a polypeptide of 377 amino acids and is considered to be the pyridoxal-P dependent histidine decarboxylase. The hdcT gene is assumed to be a histidine/histamine antiporter, and the hisRS gene is considered to be a histidyl tRNA synthetase. Recombinant Escherichia coli strains harbouring plasmids carrying the P. damselae hdc genes were shown to over-excrete histamine extracellularly. Northern blot analysis and quantitative RT-PCR revealed high levels of mono- and bi-cistronic transcripts of hdcA, hdcT and hisRS genes under conditions of low pH and histidine excess. CONCLUSIONS: The hdcA gene of P. damselae was constructed as an operon with putative histidine/histamine antiporter and histidyl-tRNA synthetase. Mono- and poly-cistronic transcripts and acid induction were detected. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of cloning the histidine decarboxylase gene cluster in gram-negative bacteria. Also, these genes were induced under acidic conditions and in the presence of excess histidine. PMID- 19302298 TI - Quantitative detection of Helicobacter pylori in water samples by real-time PCR amplification of the cag pathogenicity island gene, cagE. AB - AIMS: A new real-time PCR assay that simultaneously amplifies a 102-bp fragment of the cagE gene from Helicobacter pylori and a new internal positive control containing a specific sequence of the gyrB gene from Aeromonas hydrophila, was developed and validated for the detection of H. pylori in environmental samples. METHODS AND RESULTS: The specificity, limits of detection and quantification, repeatability, reproducibility, and accuracy of the method were calculated. The resulting values confirmed the applicability of the method for the quantitative detection of H. pylori. The feasibility of the method was also evaluated by testing 13 pyloric antrum-positive biopsies and 69 water samples, including potable (10), surface (19) and wastewater (40) matrices. The results showed that all the biopsies and 3 of the 40 wastewater samples analysed were positive. CONCLUSIONS: This real-time PCR method provides a sensitive, specific, and accurate method for the rapid quantification of H. pylori in environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCR diagnostic system proposed in this work, provides a suitable tool for the quantitative detection of H. pylori in environmental samples and can be useful for verifying the role of water as a potential route of its transmission. PMID- 19302299 TI - Carbamoylphosphate synthetase activity is essential for the optimal growth of Streptococcus thermophilus in milk. AB - AIM: The aim of the study was to study the role of carbon dioxide metabolism in Streptococcus thermophilus through investigation of the phenotype of a carbamoylphosphate synthetase-negative mutant. METHODS AND RESULTS: The effect of carbon dioxide on the nutritional requirements of Strep. thermophilus DSM20617(T) and its derivative, carbamoylphosphate synthetase-negative mutant A17(DeltacarB), was investigated by cultivating the strain in a chemically defined medium under diverse gas compositions and in milk. The results obtained revealed that CO(2) depletion or carB gene inactivation determined the auxotrophy of Strep. thermophilus for l-arginine and uracil. In addition, the parent strain grew faster than the mutant, even when milk was supplemented with uracil or arginine. CONCLUSIONS: Milk growth experiments underlined that carbamoylphosphate synthetase activity was essential for the optimal growth of Strep. thermophilus in milk. SIGNIFICANCE AND IMPACT OF THE STUDY: The study of the carbon dioxide metabolism in Strep. thermophilus revealed new insights with regard to the metabolism of this species, which could be useful for the optimization of dairy fermentation processes. PMID- 19302300 TI - Role of autoinducer-2 on the adhesion ability of Lactobacillus acidophilus. AB - AIMS: Lactobacilli adhere to the intestinal epithelium and this intimate association likely promotes retention in the gastrointestinal tract and communication with the immune system. The aim of this study was to investigate whether or not the quorum-sensing signalling molecule, autoinducer (AI)-2, was produced by Lactobacillus acidophilus and affected adherence to intestinal epithelial cells. METHODS: Microarray analysis of concentrated cells of L. acidophilus NCFM revealed several genes involved in a classic stress response and potentially adhesion. Putative genes linked to the synthesis of the interspecies signalling molecule, AI-2, were overexpressed. Examination of the NCFM genome revealed the complete pathway for AI-2 synthesis. AI-2 activity from NCFM was detected using the Vibrio harveyi BB170 assay system. Using site-specific integration, an isogenic mutation was created in luxS and the resulting mutant did not produce AI-2. In addition to some minor metabolic effects, the luxS mutation resulted in 58% decrease in adherence to Caco-2 cells. CONCLUSION: L. acidophilus NCFM encodes the genes for synthesis of the quorum-sensing signal, AI 2, and produces this molecule during planktonic growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability to produce AI-2 affects the ability of L. acidophilus to attach to intestinal epithelial cells. PMID- 19302301 TI - Isolation and partial characterization of novel genes encoding acidic cellulases from metagenomes of buffalo rumens. AB - AIMS: To clone and characterize genes encoding novel cellulases from metagenomes of buffalo rumens. METHODS AND RESULTS: A ruminal metagenomic library was constructed and functionally screened for cellulase activities and 61 independent clones expressing cellulase activities were isolated. Subcloning and sequencing of 13 positive clones expressing endoglucanase and MUCase activities identified 14 cellulase genes. Two clones carried two gene clusters that may be involved in the degradation of polysaccharide nutrients. Thirteen recombinant cellulases were partially characterized. They showed diverse optimal pH from 4 to 7. Seven cellulases were most active under acidic conditions with optimal pH of 5.5 or lower. Furthermore, one novel cellulase gene, C67-1, was overexpressed in Escherichia coli, and the purified recombinant enzyme showed optimal activity at pH 4.5 and stability in a broad pH range from pH 3.5 to 10.5. Its enzyme activity was stimulated by dl-dithiothreitol. CONCLUSIONS: The cellulases cloned in this work may play important roles in the degradation of celluloses in the variable and low pH environment in buffalo rumen. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided evidence for the diversity and function of cellulases in the rumen. The cloned cellulases may at one point of time offer potential industrial applications. PMID- 19302302 TI - The role of GAP1 gene in the nitrogen metabolism of Saccharomyces cerevisiae during wine fermentation. AB - AIM: The aim of this study was to analyse the relevance of the general amino acid permease gene (GAP1) of the wine yeast Saccharomyces cerevisiae on nitrogen metabolism and fermentation performance. METHODS AND RESULTS: We constructed a gap1 mutant in a wine strain. We compared fermentation rate, biomass production and nitrogen consumption between the gap1 mutant and its parental strain during fermentations with different nitrogen concentrations. The fermentation capacity of the gap1 mutant strain was impaired in the nitrogen-limited and -excessive conditions. The nitrogen consumption rate between the wild strain and the mutant was different for some amino acids, especially those affected by nitrogen catabolite repression (NCR). The deletion of GAP1 gene also modified the gene expression of other permeases. CONCLUSIONS: The Gap1 permease seems to be important during wine fermentations with low and high nitrogen content, not only because of its amino acid transporter role but also because of its function as an amino acid sensor. SIGNIFICANCE AND IMPACT OF THE STUDY: A possible biotechnological advantage of a gap1 mutant is its scarce consumption of arginine, whose metabolism has been related to the production of the carcinogenic ethyl carbamate. PMID- 19302303 TI - Sequential secretion of collagenolytic, elastolytic, and keratinolytic proteases in peptide-limited cultures of two Bacillus cereus strains isolated from wool. AB - AIMS: To characterize the secretion of proteolytic activities against keratin, collagen and elastin in liquid cultures of Bacillus cereus IZ-06b and IZ-06r isolated from wool. METHODS AND RESULTS: Growth of B. cereus IZ-06b and IZ-06r were characterized in batch culture. Both strains needed an organic nitrogen source, were able to grow on wool or peptone as sole carbon and nitrogen sources, and metabolized glucose, maltose and other simple sugars. Proteolytic activities were investigated in batch cultures grown in peptide-restricted, carbon sufficient medium. Secretion of proteases was induced by peptide limitation while different proteolytic activities appeared sequentially in the growth medium. When the most available components of the peptone were depleted, collagenolytic and elastolytic proteases were produced. These were later replaced by the production of keratinolytic protease. CONCLUSIONS: B. cereus can adjust its proteolytic affinity profile in response to the supply of organic nitrogen and sequentially secrete proteases with activities targeted against increasingly inaccessible proteinous substrates as the nutritional availability in the environment deteriorates. SIGNIFICANCE AND IMPACT OF THE STUDY: Peptide-limited, carbon sufficient growth media containing no proteinous substrates are well suited for protease production in B. cereus while growth conditions can be adjusted to optimize the proteolytic affinity profiles. PMID- 19302304 TI - Use of induction promoters to regulate hyaluronan synthase and UDP-glucose-6 dehydrogenase of Streptococcus zooepidemicus expression in Lactococcus lactis: a case study of the regulation mechanism of hyaluronic acid polymer. AB - AIMS: To determine the effects of the ratios of hyaluronan synthase expression level to precursor sugar UDP-GlcA biosynthesis ability on the molecular weight (MW) of hyaluronic acid (HA) in recombinant Lactococcus lactis. METHODS AND RESULTS: The genes szHasA (hyaluronan synthase gene) and szHasB (UDP-glucose-6 dehydrogenase gene) of Streptococcus zooepidemicus were introduced into L. lactis under the control of nisA promoter and lacA promoter respectively, resulting in a dual-plasmid controlled expression system. The effects of the ratios of hyaluronan synthase expression level to the precursor sugar UDP-GlcA biosynthesis ability under different induction concentration collocations with nisin and lactose on the MW of HA in recombinant L. lactis were determined. The results showed that the final weight-average molecular weight () of HA correlated with the relative ratios of HasA (hyaluronan synthase) expression level to the concentration of UDP-GlcA. CONCLUSIONS: Regulating the relative ratios of HasA expression level to the precursor sugar biosynthesis ability was an efficient method to control the size of HA. SIGNIFICANCE AND IMPACT OF THE STUDY: This study put forward a guide to establish an efficacious way to control the size of HA in fermentation. PMID- 19302305 TI - In vitro and in vivo antagonism of actinomycetes isolated from Moroccan rhizospherical soils against Sclerotium rolfsii: a causal agent of root rot on sugar beet (Beta vulgaris L.). AB - AIMS: To evaluate the ability of the isolated actinomycetes to inhibit in vitro plant pathogenic fungi and the efficacy of promising antagonistic isolates to reduce in vivo the incidence of root rot induced by Sclerotium rolfsii on sugar beet. METHODS AND RESULTS: Actinomycetes isolated from rhizosphere soil of sugar beet were screened for antagonistic activity against a number of plant pathogens, including S. rolfsii. Ten actinomycetes out of 195 screened in vitro were strongly inhibitory to S. rolfsii. These isolates were subsequently tested for their ability to inhibit sclerotial germination and hyphal growth of S. roflsii. The most important inhibitions were obtained by the culture filtrate from the isolates J-2 and B-11, including 100% inhibition of sclerotial germination and 80% inhibition of hyphal growth. These two isolates (J-2 and B-11) were then screened for their ability to protect sugar beet against infection of S. rolfsii induced root rot in a pot trial. The treatment of S. rolfsii infested soil with a biomass and culture filtrate mixture of the selected antagonists reduced significantly (P < or = 0.05) the incidence of root rot on sugar beet. Isolate J 2 was most effective and allowed a high fresh weight of sugar beet roots to be obtained. Both antagonists J-2 and B-11 were classified as belonging to the genus Streptomyces species through morphological and chemical characteristics as well as 16S rDNA analysis. CONCLUSION: Streptomyces isolates J-2 and B-11 showed a potential for controlling root rot on sugar beet and could be useful in integrated control against diverse soil borne plant pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation showed the role, which actinomycete bacteria can play to control root rot caused by S. rolfsii, in the objective to reduce treatments with chemical fungicides. PMID- 19302306 TI - Host range and lytic capability of four bacteriophages against bovine and clinical human isolates of Shiga toxin-producing Escherichia coli O157:H7. AB - AIMS: To evaluate host range and lytic capability of four bacteriophages (rV5, wV7, wV8 and wV11) against Escherichia coli O157:H7 (STEC O157:H7) from cattle and humans. METHODS AND RESULTS: Four hundred and twenty-two STEC O157:H7 isolates (297 bovine; 125 human) were obtained in Alberta, Canada. The four phages were serially diluted and incubated for 5 h with overnight cultures of STEC O157:H7 to estimate their multiplicity of infection (MOI). All bovine STEC O157:H7 were subjected to pulsed-field gel electrophoresis (PFGE) and phage typing (PT). Phage wV7 lysed all human and bovine isolates irrespective of PFGE genotype or PT phenotype and exhibited the lowest MOI (0.004-0.006, P < 0.0001) of all phages. Phages rV5 and wV11 exhibited a lower MOI (0.002-0.04, P < 0.0001) than did phage wV8 (25-29) and they had a narrower host range than wV7 or wV8. Phages rV5, wV11 and wV8 lysed 342 (81.0%), 321 (76.1%) and 407 (96.4%), respectively, of the 422 isolates. Susceptibility of bovine STEC O157:H7 to rV5, w11 and wV8 was influenced by PFGE genotype and/or PT phenotype. CONCLUSIONS: Phages exhibited activity against the majority of bovine and human STEC O157:H7 isolates. PFGE genotype and/or PT phenotype of the host-target influenced their vulnerability to phage attack. Susceptibility of bovine STEC O157:H7 to phage may also differ among farms. Both lytic capability and host range should be considered in the selection of therapeutic phage for on-farm control of STEC O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: The present work indicates that a four-phage cocktail should be equally effective at mitigating STEC O157:H7 isolates both of bovine and of human origin. Given that some STEC O157:H7 exhibited resistance to some but not all phages, a phage cocktail is the logical approach to efficacious on-farm therapy. PMID- 19302307 TI - Comparison of sampling methods to detect Salmonella infection of turkey flocks. AB - AIMS: To compare the efficiency of various sampling methods for detection of Salmonella in turkey flocks. METHODS AND RESULTS: In a field study that compared various sampling methods one pair of boot swabs taken from the whole turkey house provided suitably sensitive results for fattening and rearing flocks and was no less sensitive than two pairs, each from half the house, tested as a pooled sample. The sensitivity was further enhanced by adding a dust sample. The dust sample appeared to be particularly useful in flocks with a low prevalence, especially in breeding flocks, and was more sensitive than a method which used five pairs of boot swabs per flock. Combined incubation of a boot swab and a dust sample showed no interference between the two sample types and a maximum sensitivity of detection. Litter samples and commercial sponge drag swabs provided a lower level of detection. CONCLUSIONS: A single pair of boot swabs taken from the whole house is recommended for routine sampling of commercial rearing or fattening flocks. An additional dust sample could be added to increase detection in flocks with a low prevalence or in breeding flocks, but adding an additional pair of boot swabs would not increase detection compared with a single pair. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that significant efficiencies can be made in sampling programmes for detection of Salmonella in turkey flocks without detracting from the sensitivity. Similar studies are recommended for other poultry sectors, particularly in chicken breeding flocks. PMID- 19302308 TI - Multiplex PCR assay for simultaneous detection and differentiation of Mycobacterium tuberculosis, Mycobacterium avium complexes and other Mycobacterial species directly from clinical specimens. AB - AIMS: Polymerase chain reaction (PCR) is the most rapid and sensitive method for diagnosing mycobacterial infections and identifying the aetiological Mycobacterial species in order to administer the appropriate therapy and for better patient management. METHODS AND RESULTS: Two hundred and thirty-five samples from 145 clinically suspected cases of tuberculosis were processed for the detection of Mycobacterial infections by ZN (Ziehl Neelsen) smear examination, L-J & BACTEC MGIT-960 culture and multiplex PCR tests. The multiplex PCR comprised of genus-specific primers targeting hsp65 gene, Mycobacterium tuberculosis complex-specific primer targeting cfp10 (Rv3875, esxB) region and Mycobacterium avium complex-specific primer pairs targeting 16S-23S Internal Transcribed Spacer sequences. The multiplex PCR developed had an analytical sensitivity of 10 fg (3-4 cells) of mycobacterial DNA. The multiplex PCR test showed the highest (77.24%) detection rate, while ZN smear examination had the lowest (20%) detection rate, which was bettered by L-J culture (34.4%) and BACTEC MGIT-960 culture (50.34%) methods. The mean isolation time for M. tuberculosis was 19.03 days in L-J culture and 8.7 days in BACTEC MGIT-960 culture. Using the multiplex PCR, we could establish M. tuberculosis + M. avium co-infection in 1.3% HIV-negative and 2.9% HIV-positive patients. The multiplex PCR was also highly useful in diagnosing mycobacteraemia in 38.09% HIV-positive and 15.38% HIV negative cases. CONCLUSIONS: The developed in-house multiplex PCR could identify and differentiate the M. tuberculosis and M. avium complexes from other Mycobacterial species directly from clinical specimens. SIGNIFICANCE AND IMPACT OF THE STUDY: The triplex PCR developed by us could be used to detect and differentiate M. tuberculosis, M. avium and other mycobacteria in a single reaction tube. PMID- 19302309 TI - Factors affecting the attachment of micro-organisms isolated from ultrafiltration and reverse osmosis membranes in dairy processing plants. AB - AIMS: To identify the types of micro-organisms involved in the formation of biofilms on dairy ultrafiltration and reverse osmosis membranes and investigate factors affecting the attachment of those isolates. METHODS AND RESULTS: Micro organisms isolated from industrial membranes following standard cleaning were identified using the API culture identification system. Thirteen different isolates representing eight genera were isolated and their ability to attach to surfaces was compared using a microtitre plate assay. Three Klebsiella strains attached best, while mixed strains of Pseudomonas and Klebsiella attached better than individual strains. Whey enhanced the attachment of the isolates. The micro organisms were characterized according to cell surface hydrophobicity using the microbial adhesion to hydrocarbon (MATH) test, and cell surface charge by measuring the zeta potential. These cell surface characteristics did not show a clear relationship with the attachment of our strains. CONCLUSIONS: A variety of different micro-organisms is associated with dairy ultrafiltration and reverse osmosis membranes after cleaning, suggesting several possible sources of contamination. The cleaning of these membranes may be inadequate. The attachment of the different isolates is highly variable and enhanced in the presence of whey. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of persistent microflora colonizing dairy membrane systems will help develop strategies to mitigate biofilm development in this environment, improving hygiene in membrane processing plants. PMID- 19302310 TI - Characterization of the germination of Bacillus megaterium spores lacking enzymes that degrade the spore cortex. AB - AIMS: To determine roles of cortex lytic enzymes (CLEs) in Bacillus megaterium spore germination. METHODS AND RESULTS: Genes for B. megaterium CLEs CwlJ and SleB were inactivated and effects of loss of one or both on germination were assessed. Loss of CwlJ or SleB did not prevent completion of germination with agents that activate the spore's germinant receptors, but loss of CwlJ slowed the release of dipicolinic acid (DPA). Loss of both CLEs also did not prevent release of DPA and glutamate during germination with KBr. However, cwlJ sleB spores had decreased viability, and could not complete germination. Loss of CwlJ eliminated spore germination with Ca2+ chelated to DPA (Ca-DPA), but loss of CwlJ and SleB did not affect DPA release in dodecylamine germination. CONCLUSIONS: CwlJ and SleB play redundant roles in cortex degradation during B. megaterium spore germination, and CwlJ accelerates DPA release and is essential for Ca-DPA germination. The roles of these CLEs are similar in germination of B. megaterium and Bacillus subtilis spores. SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that redundant roles of CwlJ and SleB in cortex degradation during germination are similar in spores of Bacillus species; consequently, inhibition of these enzymes will prevent germination of Bacillus spores. PMID- 19302311 TI - Bacterial conversion of secoisolariciresinol and anhydrosecoisolariciresinol. AB - AIMS: It has been investigated whether secoisolariciresinol (SECO) and anhydrosecoisolariciresinol (AHS), an acid degradation product of SECO, could be fermented in a similar way, and to a similar extent, by members of the intestinal microbiota. METHODS AND RESULTS: AHS and SECO were demethylated by Peptostreptococcus productus, Eubacterium limosum and Clostridium methoxybenzovorans. These bacteria have been identified as members of the human intestinal flora or closely related species. Demethylated AHS and demethylated SECO were purified by preparative RP-HPLC, and subsequently subjected to fermentation with Eggerthella lenta, Clostridium scindens and Clostridium hiranonis. Eggerthella lenta efficiently dehydroxylated demethylated SECO to enterodiol, whereas the other bacteria showed no dehydroxylation activity. CONCLUSIONS: The conversion of the diol structure of SECO into the furan ring in AHS did not influence the demethylation capability of the tested bacteria. The results also showed that the extent of dehydroxylation of demethylated AHS was much lower than that of demethylated SECO. SIGNIFICANCE AND IMPACT OF THE STUDY: Plant lignans are converted into bioactive mammalian lignans by the human intestinal bacteria. This study showed that the modification of plant lignans resulted in the formation a new type of mammalian lignan. PMID- 19302312 TI - The role of biofilms and protozoa in Legionella pathogenesis: implications for drinking water. AB - Current models to study Legionella pathogenesis include the use of primary macrophages and monocyte cell lines, various free-living protozoan species and murine models of pneumonia. However, there are very few studies of Legionella spp. pathogenesis aimed at associating the role of biofilm colonization and parasitization of biofilm microbiota and release of virulent bacterial cell/vacuoles in drinking water distribution systems. Moreover, the implications of these environmental niches for drinking water exposure to pathogenic legionellae are poorly understood. This review summarizes the known mechanisms of Legionella spp. proliferation within Acanthamoeba and mammalian cells and advocates the use of the amoeba model to study Legionella pathogenicity because of their close association with Legionella spp. in the aquatic environment. The putative role of biofilms and amoebae in the proliferation, development and dissemination of potentially pathogenic Legionella spp. is also discussed. Elucidating the mechanisms of Legionella pathogenicity development in our drinking water systems will aid in elimination strategies and procedural designs for drinking water systems and in controlling exposure to Legionella spp. and similar pathogens. PMID- 19302313 TI - The assessment of the antibacterial and antifungal activities of aspirin, EDTA and aspirin-EDTA combination and their effectiveness as antibiofilm agents. AB - AIMS: To evaluate the antimicrobial activities of aspirin, EDTA and an aspirin EDTA (A-EDTA) combination against Pseudomonas aeruginosa, Escherichia coli and Candida albicans in planktonic and biofilm cultures. METHODS AND RESULTS: Minimal inhibitory concentrations (MIC) and minimal biocidal concentrations (MBC) were determined using twofold broth microdilution and viable counting methods, respectively. Aspirin's recorded MIC values ranged from 1.2 to 2.7 mg ml(-1). Checkerboard assay demonstrated a synergism in antimicrobial activity upon combination. Aspirin's minimal biofilm eradication concentration values (MBEC) against the established biofilms ranged between 1.35 and 3.83 mg ml(-1). A complete eradication of bacterial biofilms was achieved after a 4-h treatment with the A-EDTA combination. CONCLUSION: Both aspirin and EDTA possess broad spectrum antimicrobial activity for both planktonic and biofilm cultures. Aspirin used at the MBEC for 24 h was successful in eradicating P. aeruginosa, E. coli and C. albicans biofilms established on abiotic surfaces. Moreover, the exposure to the A-EDTA combination (4 h) effected complete bacterial biofilm eradication. SIGNIFICANCE AND IMPACT OF THE STUDY: There is a continuous need for the discovery of new antimicrobial agents. Aspirin and EDTA are 'nonantibiotic drugs', the combination of which can be used successfully to treat and eradicate biofilms established on abiotic surfaces. PMID- 19302314 TI - Phenotypic variation in Lactococcus lactis subsp. lactis isolates derived from intestinal tracts of marine and freshwater fish. AB - AIMS: We compared phenotypic characteristics of Lactococcus lactis subsp. lactis derived from different sources including the intestinal tract of marine fish and freshwater fish, and cheese starter culture. METHODS AND RESULTS: In the phylogenetic analysis based on partial 16S rRNA gene nucleotide sequences (1371 bp), freshwater fish-, marine fish- and cheese starter culture-derived strains were identical to that of L. lactis subsp. lactis previously reported. Fermentation profiles determined using the API 50 CH system were similar except for fermentation of several sugars including l-arabinose, mannitol, amygdalin, saccharose, trehalose, inulin and gluconate. The strains did have distinct levels of halotolerance: marine fish-derived strains > cheese starter-derived strain > freshwater fish-derived isolate. CONCLUSIONS: Lactococcus lactis subsp. lactis showed extensive diversity in phenotypic adaptation to various environments. The phenotypic properties of these strains suggested that L. lactis subsp. lactis strains from fish intestine have additional functions compared with the cheese starter-derived strain that has previously described. SIGNIFICANCE AND IMPACT OF THE STUDY: The unique phenotypic traits of the fish intestinal tract-derived L. lactis subsp. lactis might make them useful as a probiotics in aquaculture, and contribute to the development of functional foods and novel food additives, since the strains derived from fish intestines might have additional functions such as antibacterial activity. PMID- 19302315 TI - Fructose and glucose mediates enterotoxin production and anaerobic metabolism of Bacillus cereus ATCC14579(T). AB - AIMS: To determine the effects of carbohydrates on Bacillus cereus ATCC14579(T) anaerobic metabolism and enterotoxin production in amino acids rich medium. METHODS AND RESULTS: Bacillus cereus anaerobic growth on different carbohydrates (glucose, fructose, sucrose or glucose-fructose mixture) was examined in synthetic mMOD medium under continuous cultures (mu = 0.2 h(-1)). Fermentation end-products, flux partitioning at each key branch points of the mixed acid pathway and consumption or production of amino acids were determined. On both fructose and sucrose, ATP production was favoured via acetate production from acetyl-CoA. In addition, amino acids present in the growth medium showed significant variations with high consumption of serine and net production of glutamate and alanine on some or all sugars. Enterotoxins Hbl and Nhe production was high during growth on fructose (or mixtures involving a fructose moiety). CONCLUSIONS: Fructose was identified as a key sugar influencing anaerobic metabolism and toxin production of B. cereus. SIGNIFICANCE AND IMPACT OF THE STUDY: The physiological differences associated with the fermentation of the various carbohydrates clearly modify toxinogenesis indicating that the risk of foodborne pathogens is to some extent dependent upon the prevailing nutritional environment. PMID- 19302316 TI - Incorporating time postinoculation into a dose-response model of Yersinia pestis in mice. AB - AIMS: To develop a time-dependent dose-response model for describing the survival of animals exposed to Yersinia pestis. METHODS AND RESULTS: Candidate time dependent dose-response models were fitted to a survival data set for mice intraperitoneally exposed to graded doses of Y. pestis using the maximum likelihood estimation method. An exponential dose-response model with the model parameter modified by an inverse-power dependency of time postinoculation provided a statistically adequate fit to the experimental survival data. This modified model was verified by comparison with prior studies. CONCLUSIONS: The incorporated time dependency quantifies the expected temporal effect of in vivo bacteria growth in the dose-response relationship. The modified model describes the development of animal infectious response over time and represents observed responses accurately. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to incorporate time in a dose-response model for Y. pestis infection. The outcome may be used for the improved understanding of in vivo bacterial dynamics, improved postexposure decision making or as a component to better assist epidemiological investigations. PMID- 19302317 TI - Modification of azo dyes by lactic acid bacteria. AB - AIM: The ability of Lactobacillus casei and Lactobacillus paracasei to modify the azo dye, tartrazine, was recently documented as the result of the investigation on red coloured spoilage in acidified cucumbers. Fourteen other lactic acid bacteria (LAB) were screened for their capability to modify the food colouring tartrazine and other azo dyes of relevance for the textile industry. METHODS AND RESULTS: Most LAB modified tartrazine under anaerobic conditions, but not under aerobic conditions in modified chemically defined media. Microbial growth was not affected by the presence of the azo dyes in the culture medium. The product of the tartrazine modification by LAB was identified as a molecule 111 daltons larger than its precursor by liquid chromatography-mass spectrometry. This product had a purple colour under aerobic conditions and was colourless under anaerobic conditions. It absorbed light at 361 and 553 nm. CONCLUSION: LAB are capable of anabolizing azo dyes only under anaerobic conditions. IMPACT AND SIGNIFICANCE OF THE STUDY: Although micro-organisms capable of reducing the azo bond on multiple dyes have been known for decades, this is the first report of anabolism of azo dyes by food related micro-organisms, such as LAB. PMID- 19302318 TI - Effects of individual quality, reproductive success and environmental variability on survival of a long-lived seabird. AB - 1. Heterogeneity in individual quality (i.e. individuals having different performance levels that are consistent throughout life) can drive the demography of iteroparous species, but quality in the context of environmental variability has rarely been evaluated. 2. We investigated the demographic responses of a long lived seabird, the Adelie penguin (Pygoscelis adeliae), to contrasting environmental conditions as a function of reproductive success, breeding quality (BQ) and experience. A continuous index of BQ (BQI) was developed to reflect an individual's ability, relative to others, to produce viable offspring. 3. First, we assessed the relative importance of costs of reproduction vs. heterogeneity in quality by comparing survival and reproductive probabilities among deferred, successful and unsuccessful breeders under 'demanding' conditions using multistate capture-mark-recapture modelling. Then, we quantified the influence of BQI on adult survival among experienced breeders vs. the whole study population under both 'normal' and 'demanding' conditions. 4. Higher survival rates were exhibited by successful (74-76%) compared to unsuccessful breeders (64%); the former also more frequently reproduced successfully at year t + 1. 5. From 1997 to 2006, adult survival ranged from 64-79%, with BQI accounting for 91% of variability in the entire study population, but only 17% in experienced breeders. The weakened relationship between BQI and survival in experienced breeders supports the theory that selection during the first reproductive event accounts for a more homogeneous pool of experienced breeders. 6. No significant effect of environmental covariates on survival was evident, suggesting that what appeared to be demanding conditions were within the range that could be buffered by this species. 7. For the first time in seabirds, a quadratic relationship between adult survival and BQI showed that adult survival is shaped by both heterogeneity in quality and reproductive costs. Our study confirms that population demographic patterns are affected by factors at the individuals' level (e.g., individual quality) that are obscured at population-scale levels. PMID- 19302319 TI - The effects of colonization, extinction and competition on co-existence in metacommunities. AB - 1. The co-existence of competitors in heterogeneous landscapes depends on the processes of colonization, extinction and spatial scale. In this study, we explore the metapopulation dynamics of competitive interactions. 2. Rather than simply evaluating the outcome of interspecific competition in the traditional manner, we focus on both the local population dynamic effects and the regional metapopulation processes affecting species co-existence. 3. We develop a theoretical model of regional co-existence to generate a set of predictions on the patterns of colonization necessary for co-existence and the regional processes that can lead to competitive exclusion. We empirically test these predictions using metacommunity microcosms of the interaction between two bruchid beetles (Callosobruchus chinensis, Callosobruchus maculatus). 4. Using well replicated time series of the interaction between the bruchids and statistical methods of model fitting, we show how the qualitative and quantitative pattern of interspecific competition between the bruchid beetles is shaped by the structure of the metacommunity. 5. In unlimited dispersal metacommunities, the global exclusion of the inferior competitor is shown to be influenced more by the processes associated with extinction rather than low colonization ability. In restricted dispersal metacommunities, we show how the co-existence of competitors in a spatially heterogeneous habitat (patches connected through limited dispersal) is affected by Allee effects and life-history [colonization (dispersal) - competition] trade-offs. PMID- 19302320 TI - Patterns of mortality for each life-history stage in a population of the endangered New Zealand stitchbird. AB - 1. Using data from 396 breeding attempts over an 8-year period, we investigated age- and stage-specific survival rates and their modifying factors in a closed island population of the New Zealand stitchbird (or hihi, Notiomystis cincta Du Bus). 2. Survival probability generally increased over time; however, at each life-history transition, survival in the new stage started lower than at the end of the previous stage, creating a 'saw-tooth' function of age-related survival. 3. The probability of an egg hatching was low (0.73 +/- 0.01): most likely a consequence of genetic bottlenecks previously endured by this population. There was strong support for a positive relationship between hatching rate and the subsequent survival of the female parent, and hatching success declining for females > 4 years old. 4. Nestling survival probability increased as a function of brood size and days since hatching, and decreased relative to daily maximum ambient temperature and hatching date. Support for models including ambient temperature was greater than for other covariates, with the majority of this temperature-mediated survival effect being restricted to the early nestling stage. 5. Fledglings had low survival rates in the first two weeks after leaving the nest, with post-fledging survival related to the fledgling's mass. Two months after fledging, juvenile survival probability plateaued and remained relatively constant for the following autumn, winter and spring/summer breeding season. There was no effect of sex or season on adult survival probability. However, there was strong support for age-specific variation in adult survival, with survival likelihood increasing during the first four years before showing evidence of a senescence decline. 6. Within-stage survival increases were likely related to stage-specific selection pressures initially weeding out individuals of poorer phenotypes for the environment specific to each life-history stage. Such a mechanism explains the initial high mortality at life-history transitions; a well-adapted phenotype for one stage may not necessarily be so well adapted for subsequent stages. These patterns are not only valuable for examining life history theory, but also for understanding the regulation of vital rates in an endangered species and providing a basis from which better population management models and harvesting regimes can be derived. PMID- 19302321 TI - Evidence for the involvement of an alternate rodent host in the dynamics of introduced plague in prairie dogs. AB - 1. The introduction of plague to North America is a significant threat to colonies of prairie dogs (Cynomys ludovicianus), a species of conservation concern in the Great Plains. Other small rodents are exposed to the causative agent, Yersinia pestis, during or after epizootics; yet, its effect on these rodents is not known, and their role in transmitting and maintaining plague in the absence of prairie dogs remains unclear. 2. We live-trapped small rodents and collected their fleas on 11 colonies before, during and after plague epizootics in Colorado, USA, from 2004 to 2006. Molecular genetic (polymerase chain reaction) assays were used to identify Y. pestis in fleas. 3. Abundance of northern grasshopper mice (Onychomys leucogaster) was low on sites following epizootics in 2004, and declined markedly following plague onset on other colonies in 2005. These changes coincided with exposure of grasshopper mice to plague, and with periods when mice became infested with large numbers of prairie dog fleas (Oropsylla hirsuta), including some that were infected with Y. pestis. Additionally, several Pleochaetis exilis, fleas restricted to grasshopper mice and never found on prairie dogs on our site, were polymerase chain reaction positive for Y. pestis, indicating that grasshopper mice can infect their own fleas. No changes in abundance of other rodent species could be attributed to plague, and no other rodents hosted O. hirsuta during epizootics, or harboured Y. pestis-infected fleas. 4. In spring 2004, grasshopper mice were most numerous in colonies that suffered plague the following year, and the pattern of colony extinctions over a 12-year period mirrored patterns of grasshopper mouse abundance in our study area, suggesting that colonies with high densities of grasshopper mice may be more susceptible to outbreaks. We speculate that grasshopper mice help spread Y. pestis during epizootics through their ability to survive infection, harbour prairie dog fleas and, during their wide-ranging movements, transport infected fleas among burrows, which functionally connects prairie dog coteries that would otherwise be socially distinct. PMID- 19302322 TI - Sex-specific adult dispersal and its selective consequences in the brown anole, Anolis sagrei. AB - 1. Dispersal behaviour is a potentially risky life-history strategy that can impact habitat use as well as competition over territories and mates. 2. I studied natural selection on dispersal behaviour over the course of four breeding seasons in a Bahamian population of brown anoles (Anolis sagrei). 3. Both males and females showed extremely high site fidelity over the course of each reproductive season. Movement distance in males was negatively correlated with body size at first capture in spring (small males dispersed further). Moreover, differences in body size between dispersing males and the body size of the male territory residents that they replaced were correlated with the distance that males dispersed. Relatively larger males dispersed shorter distances compared with relatively smaller males. There was no relationship between dispersal distance and body size in females. However, females were more likely to disperse away from areas of low female density and into areas of higher female density, and female dispersal distances were negatively correlated with the number of female neighbours at the site of capture in spring (before dispersal). 4. These data suggest that, whereas male dispersal is driven by inter-male competition, female dispersal is more likely related to variation in territory quality. 5. Natural selection acted on dispersal distance in conjunction with male, but not female body size. Although smaller males were the more likely to disperse, these males paid a high cost in terms of viability indicating that dispersal is a potentially risky strategy. PMID- 19302323 TI - Scared fish get lazy, and lazy fish get fat. AB - 1. Many biological textbooks present predator-induced morphological changes in prey species as an example of an adaptive response, because the morphological change is associated with lower predation risk. Here we show that the adaptive morphological response observed in many systems may actually be an indirect effect of decreased activity - which reduces the predation risk - rather than a direct adaptive response. 2. One of the classical examples comes from crucian carp, where the presence of pike leads to a deeper body. We manipulated pike cues (presence and absence) and water current (standing and running water) and found that both standing water and pike cues similarly and independently induced a deeper body. 3. Since the presence of pike cues as well as standing water might be associated with low swimming activity, we suggest that the presence of pike causes a reduction in activity (antipredator behaviour). Reduced activity subsequently induces a deeper body, possibly because the energy saved is allocated to a higher growth rate. 4. Our result suggests that even if morphological change is adaptive, it might be induced indirectly via activity. This important conceptual difference may be similar in many other systems. PMID- 19302324 TI - Multidrug resistance gene deficient (mdr1a-/-) mice have an altered caecal microbiota that precedes the onset of intestinal inflammation. AB - AIM: To compare caecal microbiota from mdr1a(-/-) and wild type (FVB) mice to identify differences in the bacterial community that could influence the intestinal inflammation. METHODS AND RESULTS: Caecal microbiota of mdr1a(-/-) and FVB mice were evaluated at 12 and 25 weeks of age using denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR. DGGE fingerprints of FVB and mdr1a(-/-) mice (with no intestinal inflammation) at 12 weeks revealed differences in the presence of DNA fragments identified as Bacteroides fragilis, B. thetaiotaomicron, B. vulgatus and an uncultured alphaproteobacterium. Escherichia coli and Acinetobacter sp. were only identified in DGGE profiles of mdr1a(-/-) mice at 25 weeks (with severe intestinal inflammation), which also had a lower number of total bacteria in the caecum compared with FVB mice at same age. CONCLUSIONS: Differences found in the caecal microbiota of FVB and mdr1a(-/ ) mice (12 weeks) suggest that the lack of Abcb1 transporters in intestinal cells due to the disruption of the mdr1a gene might lead to changes in the caecal microbiota. The altered microbiota along with the genetic defect could contribute to the development of intestinal inflammation in mdr1a(-/-) mice. SIGNIFICANCE AND IMPACT OF THE STUDY: Differences in caecal microbiota of mdr1a(-/-) and FVB mice (12 weeks) suggest genotype specific colonization. The results provide evidence that Abcb1 transporters may regulate host interactions with commensal bacteria. Future work is needed to identify the mechanisms involved in this possible cross-talk between the host intestinal cells and microbiota. PMID- 19302325 TI - Leptospirosis: risks during recreational activities. AB - Summary Rats, dogs, cattle, bats and sea lions, exemplify the diversity of mammalian species that can facilitate transmission of the zoonotic disease leptospirosis. The causative agent, pathogenic species of Leptospira, is shed in urine of chronically infected hosts. Direct contact with infected urine, or indirectly with water sources contaminated with infected urine, poses a risk of infection for humans exposed during water-related recreational and occupational activities. New serovars of Leptospira and maintenance hosts continue to be identified. In the western world, incidences of recreational exposure are increasing, while incidences of occupational exposure are decreasing. Adventure travellers returning from tropical regions, are presenting at clinics with symptoms of leptospirosis following participation in high risk activities including white water rafting, triathlons, endurance races and caving. Risks of infection can be reduced with increased awareness of how the disease is contracted, by avoiding contact with high risk water sources and the use of prophylaxis during high risk activities. Molecular techniques can be used to provide risk assessments prior to competition, to supplement epidemiology, and to assess shedding of Leptospira in urine samples. PMID- 19302326 TI - Screening and identification of vip genes in Bacillus thuringiensis strains. AB - AIMS: To identify known vip genes and to detect potentially novel vip genes in a collection of 507 strains of Bacillus thuringiensis. METHODS AND RESULTS: Following a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) strategy, four restriction patterns were found within the vip1 family: vip1Aa1, vip1Ba1/vip1Ba2 and vip1Ca. In the screening of vip2 genes, patterns similar to those of vip2Aa1, vip2Ba1/vip2Ba2 and vip2Ac1 genes were observed. Patterns for vip3Aa1, vip3Ae2 and vip3Af1 were found among vip3 genes. Two new patterns revealed novel vip1 and vip3A genes. The observed frequency of genes belonging to vip1 and vip2 families was around 10%, whereas 48.9% of the strains showed amplification of vip3 genes. A tendency of vip and cry genes to occur together has been observed in this collection of B. thuringiensis strains. CONCLUSIONS: Ten different patterns of vip genes belonging to the three vip families and two novel vip genes have been identified in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time that vip1 and vip2 genes have been identified by PCR-RFLP. Furthermore, the results show that the strategy used in this study can lead to the classification of known vip genes as well as the identification of novel vip genes. PMID- 19302327 TI - Pilot- and bench-scale testing of faecal indicator bacteria survival in marine beach sand near point sources. AB - AIM: Factors affecting faecal indicator bacteria (FIB) and pathogen survival/persistence in sand remain largely unstudied. This work elucidates how biological and physical factors affect die-off in beach sand following sewage spills. METHODS AND RESULTS: Solar disinfection with mechanical mixing was pilot tested as a disinfection procedure after a large sewage spill in Los Angeles. Effects of solar exposure, mechanical mixing, predation and/or competition, season, and moisture were tested at bench scale. First-order decay constants for Escherichia coli ranged between -0.23 and -1.02 per day, and for enterococci between -0.5 and -1.0 per day. Desiccation was a dominant factor for E. coli but not enterococci inactivation. Effects of season were investigated through a comparison of experimental results from winter, spring, and fall. CONCLUSIONS: Moisture was the dominant factor controlling E. coli inactivation kinetics. Initial microbial community and sand temperature were also important factors. Mechanical mixing, common in beach grooming, did not consistently reduce bacterial levels. SIGNIFICANCE AND IMPACT OF THE STUDY: Inactivation rates are mainly dependent on moisture and high sand temperature. Chlorination was an effective disinfection treatment in sand microcosms inoculated with raw influent. PMID- 19302328 TI - Role of the alternative sigma factor sigma on Staphylococcus aureus resistance to stresses of relevance to food preservation. AB - AIMS: To examine the role of the alternative general stress sigma factor sigma(B) on the resistance of Staphylococcus aureus to stresses of relevance to food preservation, with special emphasis on emerging technologies such as pulsed electric fields (PEF) and high hydrostatic pressure (HHP). METHODS AND RESULTS: S. aureus strain Newman and its isogenic DeltasigB mutant were grown to exponential and stationary growth phases and its resistance to various stresses was tested. The absence of the sigma(B) factor caused a decrease in the resistance to heat, PEF, HHP, alkali, acid and hydrogen peroxide. In the case of heat, the influence of the sigma(B) factor was particularly important, and decreases in decimal reduction time values of ninefold were observed as a result of its deficiency. The increased thermotolerance of the parental strain as compared with the sigB mutant could be attributed to a better capacity to sustain and repair sublethal damages caused by heat. CONCLUSIONS: sigma(B) factor provides S. aureus cells with resistance to multiple stresses, increasing survival to heat, PEF and HHP treatments. SIGNIFICANCE AND IMPACT OF THE STUDY: Results obtained in this work help in understanding the physiological mechanisms behind cell survival and death in food-processing environments. PMID- 19302329 TI - Sources of psychrophilic and psychrotolerant clostridia causing spoilage of vacuum-packed chilled meats, as determined by PCR amplification procedure. AB - AIMS: To determine possible preslaughter and processing sources of psychrophilic and psychrotolerant clostridia causing spoilage of vacuum-packed chilled meats. METHODS AND RESULTS: Molecular methods based on the polymerase chain reaction (PCR) amplification of specific 16S rDNA fragments were used to detect the presence of Clostridium gasigenes, Clostridium estertheticum, Clostridium algidicarnis and Clostridium putrefaciens in a total of 357 samples collected from ten slaughter stock supply farms, slaughter stock, two lamb-processing plants, their environments, dressed carcasses and final vacuum-packed meat stored at -0.5 degrees C for 5(1/2) weeks. Clostridium gasigenes, C. estertheticum and C. algidicarnis/C. putrefaciens were commonly detected in farm, faeces, fleece and processing environmental samples collected at the slaughter floor operations prior to fleece removal, but all these micro-organisms were detected in only 4 out of 26 cooling floor and chiller environmental samples. One out of 42 boning room environmental samples tested positive for the presence of C. gasigenes and C. estertheticum, but 25 out of 42 of these samples were positive for C. algidicarnis/C. putrefaciens. Nearly all of the 31 faecal samples tested positive for the presence of C. gasigenes and C. estertheticum; however, only two of these samples were positive for C. algidicarnis and/or C. putrefaciens. Clostridial species that were subject to this investigation were frequently detected on chilled dressed carcasses. CONCLUSIONS: The major qualitative and quantitative differences between the results of PCR detection obtained with the primers specific for 'blown pack' -causing clostridia (C. gasigenes and C. estertheticum) and those obtained with primers specific for C. algidicarnis and C. putrefaciens suggest that the control of meat spoilage caused by different groups of meat clostridia is best approached individually for each group. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper provides information significant for controlling meat spoilage-causing clostridia in the meat-processing plants. PMID- 19302330 TI - Influence of lyophilization, fluidized bed drying, addition of protectants, and storage on the viability of lactic acid bacteria. AB - AIMS: The present study focuses on the impact of two different drying technologies and the influence of protectants on process survival and storage stability of the two lactic acid bacterial strains Enterococcus faecium and Lactobacillus plantarum. METHODS AND RESULTS: After incubation with the protectants glucose, sucrose, trehalose, and maltodextrin the concentrated bacterial suspensions were subjected to fluidized bed drying and lyophilization and subsequently stored at 4, 22, and 35 degrees C for half a year. Lactobacillus plantarum turned out to be more sensitive to both drying methods than Ent. faecium. Without the addition of a protectant cells of both strains suffered higher losses during fluidized bed drying. Elevated storage temperatures correlate with a higher decline of viable bacterial cells. CONCLUSIONS: Although survival rates varied between the strains, the nonreducing disaccharides revealed overall best protection for both investigated lactic acid bacteria during processing and storage. The addition of protective carbohydrates can prevent the decline in viability during fluidized bed drying. SIGNIFICANCE AND IMPACT OF THE STUDY: The influence of protectants proved to be species specific and therefore needs to be determined on a case-to-case basis. Survival rates, duration, and energy consumption appear to be the crucial parameters to evaluate the economy of production processes for industrial starter cultures. PMID- 19302331 TI - Synergistic interactions between cefalexin and kanamycin in Mueller-Hinton broth medium and in milk. AB - AIMS: This study investigated the in vitro bactericidal activity of an intramammary drug product by comparing the kill kinetics of cefalexin and kanamycin, alone and in fixed ratio combination, against Streptococcus uberis, Staphylococcus aureus and Escherichia coli strains isolated from field cases of bovine mastitis. The effect of milk as a diluent on the rate of bacterial killing was also assessed. METHODS AND RESULTS: Antibacterial kill kinetics was determined against each bacterial strain in Mueller-Hinton broth (MHB) and in milk. In MHB, the fixed cefalexin : kanamycin combination (1.5 : 1 w/w) exhibited a clear synergistic bactericidal activity against the strains tested. The combination also showed an enhanced killing activity in milk, as compared to either agent alone. CONCLUSIONS: The data show the occurrence of synergistic interactions between cefalexin and kanamycin, resulting in a faster and enhanced bactericidal activity against major mastitis pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrated that the combination exhibited a larger and faster rate of kill of S. aureus, S. uberis and E. coli compared to either cefalexin or kanamycin alone, while using a lower total amount of antibiotic. Synergistic and additive effects were also observed when milk was used as a medium. The results support the use of this combination of narrow spectrum antibiotics to treat clinical mastitis via the intramammary route and provide data on its killing kinetics. PMID- 19302332 TI - Structural and functional responses of sewage microbial communities used for the treatment of a complex mixture of volatile organic compounds (VOCs). AB - AIMS: The aim of this work was to assess the impact of the applied mass loading on the selection of an efficient microbial community able to degrade a complex mixture of volatile organic compounds (VOCs). METHODS AND RESULTS: Two reactors were used and were supplied with a gaseous effluent containing 11 VOCs with different concentrations. The response of the microflora was monitored as a function of time: biodegradation activity, bacterial density and diversity. The results showed that the applied mass loading seems to have an impact on the functioning and the genetic structure of the bacterial community. CONCLUSIONS: A high mass loading seems to induce a low efficient functioning in terms of elimination efficiency and a simplification of the genetic structure of the total bacterial community with the apparition of a dominant microflora. A low mass loading seems to favour a better functioning and allows to keep a healthier bacterial diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: In the treatment processes of gaseous effluents, it would be judicious to define the functioning parameters of the process to keep the diversity of important functional bacterial groups. These results provide also useful information about changes in microbial communities following natural or anthropogenic alterations in different ecosystems. PMID- 19302333 TI - Flexible exportation mechanisms of arthrofactin in Pseudomonas sp. MIS38. AB - AIMS: To obtain further insights into transportation mechanisms of a most effective biosurfactant, arthrofactin in Pseudomonas sp. MIS38. METHODS AND RESULTS: A cluster genes arfA/B/C encodes an arthrofactin synthetase complex (ArfA/B/C). Downstream of the arfA/B/C lie genes encoding a putative periplasmic protein (ArfD, 362 aa) and a putative ATP-binding cassette transporter (ArfE, 651 aa), namely arfD and arfE, respectively. The arfA/B/C, arfD, and arfE form an operon suggesting their functional connection. Gene knockout mutants ArfD:Km, ArfE:Km, ArfD:Tc/ArfE:Km, and gene overexpression strains MIS38(pME6032_arfD/E) and ArfE:Km(pME6032_arfD/E) were prepared and analysed for arthrofactin production profiles. It was found that the production levels of arthrofactin were temporally reduced in the mutants or increased in the gene overexpression strains, but they eventually became similar level to that of MIS38. Addition of ABC transporter inhibitors, glibenclamide and sodium ortho-vanadate dramatically reduced the production levels of arthrofactin. This excludes a possibility that arthrofactin is exported by diffusion with the aid of its own high surfactant activity. CONCLUSIONS: ArfD/E is not an exclusive but a primary exporter of arthrofactin during early growth stage. Reduction in the arthrofactin productivity of arfD and arfE knockout mutants was eventually rescued by another ABC transporter system. Effects of arfD and arfE overexpression were evident only for 1-day cultivation. Multiple ATP dependent active transporter systems are responsible for the production of arthrofactin. SIGNIFICANCE AND IMPACT OF THE STUDY: Pseudomonas bacteria are characterized to be endued with multiple exporter and efflux systems for secondary metabolites including antibiotics, plant toxins, and biosurfactants. The present work demonstrates exceptionally flexible and highly controlled transportation mechanisms of a most effective lipopeptide biosurfactant, arthrofactin in Pseudomonas sp. MIS38. Because lipopeptide biosurfactants are known to enhance efficacy of bioactive compounds and arfA/B/C/D/E orthologous genes are also found in plant pathogenic P. fluorescens and P. syringae strains, the knowledge would also contribute to develop a technology controlling plant diseases. PMID- 19302334 TI - Detection of infectious rotavirus in naturally contaminated source waters for drinking water production. AB - AIMS: To assess public health risks of rotavirus via drinking water consumption, a cell culture-PCR assay was developed and optimized for the detection of infectious environmental rotavirus strains in naturally contaminated source waters for drinking water production. METHODS AND RESULTS: Infectious rotavirus concentrations were estimated by an optimized cell culture-PCR assay as most probable numbers by using the presence or absence of replicated virus in different sample volumes. Infectious rotavirus was detected in 11 of 12 source water samples in concentrations varying from 0.19 (0.01-0.87) to 8.3 (1.8-34.0) infectious PCR detectable units per litre (IPDU/l), which was not significantly different from the concentrations of infectious enterovirus in these samples. CONCLUSIONS: In 55% of the samples, rotavirus genomes were 1000 to 10 000 times (3 log(10)-4 log(10)) more abundantly present than infectious rotavirus particles, whereas in the remaining 45% of the samples, rotavirus genomes were less than 1000 times (<3 log(10)) more abundantly present. SIGNIFICANCE AND IMPACT OF THE STUDY: The broad variation observed in the ratios of rotavirus RNA and infectious particles demonstrates the importance of detecting infectious viruses instead of viral RNA for the purposes involving estimations of public health risks. PMID- 19302335 TI - Hepatitis B-related hepatocellular carcinoma: epidemiological characteristics and disease burden. AB - Worldwide, 350 million people are chronically infected with hepatitis B virus (HBV) who are at greater risk of hepatocellular carcinoma (HCC) compared with uninfected people. The relative risks of HCC among people infected with HBV ranges from 5 to 49 in case-control studies and from 7 to 98 in cohort studies. More than 50% of HCC cases worldwide and 70-80% of HCC cases in highly HBV endemic regions are attributable to HBV. Incidence of HCC (per 100,000 person/year) among people with chronic HBV infection ranges from 400 to 800 in male and from 120 to 180 in female. Factors associated with increased risk of HCC include demographic characteristics (male sex and older age), lifestyles (heavy alcohol consumption and smoking), viral factors (genotype C, D F, high level of HBV DNA, core/precore mutation) and clinical factors (cirrhosis, elevated alpha fetoprotein (AFP) and alanine aminotransferase (ALT)). HBV-related HCC has extremely poor prognosis with median survival less than 16 months. Survival rates of HBV-related HCC ranged from 36% to 67% after 1 year and from 15% to 26% after 5 year of diagnosis. Older age, liver function impairment, vascular invasion, tumour aggressiveness and elevated AFP are associated with HCC survival. Global burden of HBV-related liver disease is still a major challenge for public health in the 21st century. While decreases in incidence of HBV infection have been observed in birth cohorts following the introduction of universal infant HBV vaccination programme, HBV-related HCC incidence in is projected to increase for at least two decades because of the high prevalence of chronic HBV infection and prolonged latency to HCC development. To reduce HBV-related HCC continued expansion of universal infant HBV vaccination is required along with antiviral therapy targeted to those individuals at highest risk of HCC. Broad public health strategies should include routine testing to identify chronic HBV infection, improved health infrastructures including human resource to provide diagnosis and treatment assessment. PMID- 19302336 TI - Mutational analysis of the hepatitis C virus E1 glycoprotein in retroviral pseudoparticles and cell-culture-derived H77/JFH1 chimeric infectious virus particles. AB - Cell entry by enveloped viruses is mediated by viral glycoproteins, and generally involves a short hydrophobic peptide (fusion peptide) that inserts into the cellular membrane. An internal hydrophobic domain within E1 (aa262-290) of hepatitis C virus (HCV) may function as a fusion peptide. Retrovirus-based HCV pseudotyped viruses (HCVpp; genotype 1a) containing Ala or Pro substitutions at conserved amino acid positions within this putative fusion peptide were generated. Mutation of conserved residues significantly reduced efficiency of HCVpp entry into Huh-7 cells. The majority of amino acid substitutions appeared to disrupt necessary interactions between E1 and E2. For some mutants, reductions in HCVpp-associated E1 were associated with the incorporation of a high molecular weight, hyperglycosylated E2 that displayed decreased CD81-binding. Other entry deficient mutants displayed normal E1E2 incorporation into pseudoparticles and normal CD81-binding, and therefore might affect viral fusion. One mutant (S283P) consistently displayed two- to threefold higher infectivity than did wild-type. Three mutations that decreased HCVpp infectivity also reduced levels of HCVcc infectious virus production. However, the S283P mutation had a different effect in the two systems as it did not increase production of infectious HCVcc. This comprehensive mutational analysis of the putative HCV fusion peptide provides insight into the role of E1 in its interaction with E2 and in HCV entry. PMID- 19302337 TI - Low interleukin-10 production by monocytes of patients with a self-limiting hepatitis C virus infection. AB - Host factors seem to be crucial for the spontaneous clearance of hepatitis C virus (HCV). Monocytes play a pivotal role in innate immunity and help regulate adaptive responses. This study assesses the characteristics of monocytes from patients with self-limiting HCV infections. We studied 35 consecutive patients [11 with a self-limiting HCV infection, 16 chronically infected with HCV and sustained virological responders (SVR) following antiviral therapy, and eight chronically infected HCV but untreated] and eight healthy donors (HD). The production of interleukin (IL)-10, tumour necrosis factor-alpha (TNF-alpha) and IL-12p40 by monocytes stimulated with lipopolysaccharides(LPS) or HCV Core protein was measured by enzyme-linked immunoassay. Monocyte surface markers were analysed by flow cytometry. LPS and Core protein triggered IL-10 and TNF-alpha production, but monocytes from self-limiting infection patients produced significantly less IL-10 and TNF-alpha than those of SVR, chronically infected or HD (P < 0.05), while IL-12p40 production was unchanged. This cytokine production profile did not appear to be due to expansion of the CD14(+) CD16(+) monocyte subset or to a classical or alternative activation monocyte profile. Monocytes from self-limiting infection patients had more CCR7 than those from SVR or chronically infected patients (P < 0.05). Monocytes of self-limiting infection patients appear to produce little IL-10 and TNF-alpha in response to viral or unspecific stimulation and to have a higher CCR7 expression. This profile seems to be independent to a particular monocyte subset or activation state. Low IL-10 production may help establish an effective immune response and spontaneous HCV clearance. PMID- 19302338 TI - Patients achieving clearance of HCV with interferon therapy recover from decreased retinol-binding protein 4 levels. AB - Retinol-binding protein 4 (RBP4) is a recently identified adipokine that is elevated in the blood in several insulin-resistant states. We investigated the association between plasma RBP4 and histological and biochemical characteristics of chronic hepatitis C (CHC), as well as changes in RBP4 levels following interferon therapy. Eighty-one patients with CHC infected with genotype 1 received treatment with peginterferon plus ribavirin. Histological data were available for 41 out of 81 patients before treatment, and the degree of fibrosis, inflammation and steatosis was assessed. Plasma levels of RBP4 were determined in serial samples (before, at the end of treatment, and at 6 months post-treatment). RBP4 levels were lower in CHC patients than in control subjects (34.6 +/- 12.3 microg/mL vs 46.2 +/- 10.5 microg/mL; P 1.5) groups. RESULTS: The authors did not identify any randomized controlled trials (RCT), but included four observational studies (three retrospective registry and one prospective cohort studies), which enrolled 1,511 patients cumulatively. One study found a statistically significant difference in mortality rate, favoring high FFP:PRBC ratio (relative risk = 0.72, 95% confidence interval [CI] = 0.59 to 0.89), while three studies showed no benefits. One study reported higher rates of sepsis and single/multiorgan failure (MOF), and another study revealed a higher risk of nosocomial infections and acute respiratory distress syndrome (ARDS) in the high FFP:PRBC ratio group. CONCLUSION: Three retrospective registry reviews with suboptimal methodologies and one prospective cohort study provide inadequate evidence to support or refute the use of a high FFP:PRBC ratio in patients with severe trauma. PMID- 19302365 TI - Health beliefs toward cardiovascular risk reduction in patients admitted to chest pain observation units. AB - OBJECTIVES: Even after acute coronary syndrome (ACS) is ruled out, observational studies have suggested that many patients with nonspecific chest pain have a high burden of cardiovascular risk factors (CRFs) and are at increased long-term risk of ischemic heart disease (IHD)-related mortality. The aim of this study was to evaluate the premise that evaluation in an observation unit for symptoms of possible ACS is a "teachable moment" with regard to modification of CRFs. METHODS: The authors conducted a baseline face-to-face interview and a 3-month telephone interview of 83 adult patients with at least one modifiable CRF who presented with symptoms of possible ACS to an academic medical center. Existing questionnaires were adapted to measure Health Belief Model (HBM) constructs for IHD. Stage of change and self-reported CRF-related behaviors (diet, exercise, and smoking) were assessed using previously validated measures. The paired t-test or signed rank test was used to compare baseline and 3-month measures of health behavior within the analysis sample. RESULTS: Of the 83 study patients, 45 and 40% reported having received clinician advice regarding diet and physical activity during the observation unit encounter, respectively; 69% of current smokers received advice to quit smoking. Patients reported lower susceptibility to IHD (13.3 vs. 14.0, p = 0.06) and greater perceived benefit of healthy lifestyles (27.5 vs. 26.4, p = 0.0003) at 3-month follow-up compared to baseline. Patients also reported greater readiness to change and improved self-reported behaviors at follow-up (vs. baseline): decreased intake of saturated fat (10.1% vs. 10.5% of total calories, p = 0.005), increased fruit and vegetable intake (4.0 servings/day vs. 3.6 servings/day, p = 0.01), and fewer cigarettes (13 vs. 18, p = 0.002). CONCLUSIONS: Observed changes in IHD health beliefs and CRF related behaviors during follow-up support the idea that observation unit admission is a teachable moment. Patients with modifiable risk factors may benefit from systematic interventions to deliver CRF-related counseling during observation unit evaluation. PMID- 19302366 TI - Prehospital ultrasound diagnosis of traumatic pericardial effusion. PMID- 19302368 TI - Emergent management of anterior epistaxis. PMID- 19302369 TI - Detection of undiagnosed diabetes and prediabetic states in high-risk emergency department patients. AB - BACKGROUND: Diabetes is often not diagnosed until complications appear, and one third of those with diabetes may be undiagnosed. Prediabetes and diabetes are conditions in which early detection would be appropriate, because the duration of hyperglycemia is a predictor of adverse outcomes, and there are effective interventions to prevent disease progression and to reduce complications. OBJECTIVES: The objectives were to determine the prevalence of diabetes mellitus and prediabetes in emergency department (ED) patients with an elevated random glucose or risk factors for diabetes but without previously diagnosed diabetes and to identify which at-risk ED patients should be considered for referral for confirmatory diagnostic testing. METHODS: This two-part study was composed of a prospective 2-year cohort study, and a 1-week cross-sectional survey substudy, set in an urban ED in Los Angeles County, California. A convenience sample was enrolled of 528 ED patients without previously diagnosed diabetes with either 1) a random serum glucose > or = 140 mg/dL regardless of the time of last food intake or a random serum glucose > or = 126 mg/dL if more than 2 hours since last food intake or 2) at least two predefined diabetes risk factors. Measurements included presence of diabetes risk factors, ED glucose, cortisol, insulin and glycosylated hemoglobin (HbA(1c)), and 2-hour oral glucose tolerance test results, administered at 6-week follow-up. RESULTS: Glycemic status was confirmed at follow-up in 256 (48%) of the 528 patients. Twenty-seven (11%) were found to have diabetes, 141 (55%) had prediabetes, and 88 (34%) had normal results. Age, ED glucose, HbA(1c), cortisol, and random serum glucose > or = 140 mg/dL were associated with both diabetes and prediabetes on univariate analysis. A random serum glucose > or = 126 mg/dL after 2 hours of fasting was associated with diabetes but not prediabetes; ED cortisol, insulin, age > or = 45 years, race, and calculated body mass index (BMI) were associated with prediabetes but not diabetes. In multivariable models, among factors measurable in the ED, the only independent predictor of diabetes was ED glucose, while ED glucose, age > or = 45 years, and symptoms of polyuria and polydipsia were independent predictors of prediabetes. All at-risk subjects with a random ED blood glucose > 155 mg/dL had either prediabetes or diabetes on follow-up testing. CONCLUSIONS: A substantial fraction of this urban ED study population was at risk for undiagnosed diabetes and prediabetes, and among the at-risk patients referred for follow-up, the majority demonstrated diabetes or prediabetes. Notably, all patients with two risk factors and a random serum glucose > 155 mg/dL were later diagnosed with prediabetes or diabetes. Consideration should be given to referring ED patients with risk factors and a random glucose > 155 mg/dL for follow-up testing. PMID- 19302370 TI - Reduced expression of alpha-1,2-mannosidase I extends lifespan in Drosophila melanogaster and Caenorhabditis elegans. AB - Exposure to sub-lethal levels of stress, or hormesis, was a means to induce longevity. By screening for mutations that enhance resistance to multiple stresses, we identified multiple alleles of alpha-1,2-mannosidase I (mas1) which, in addition to promoting stress resistance, also extended longevity. Longevity enhancement is also observed when mas1 expression is reduced via RNA interference in both Drosophila melanogaster and Caenorhabditis elegans. The screen also identified Edem1 (Edm1), a gene downstream of mas1, as a modulator of lifespan. As double mutants for both mas1 and Edm1 showed no additional longevity enhancement, it appeared that both mutations function within a common pathway to extend lifespan. Molecular analysis of these mutants revealed that the expression of BiP, a putative biomarker of dietary restriction (DR), is down-regulated in response to reductions in mas1 expression. These findings suggested that mutations in mas1 may extend longevity by modulating DR. PMID- 19302371 TI - Race/ethnicity and telomere length in the Multi-Ethnic Study of Atherosclerosis. AB - Telomere length has emerged as a marker of exposure to oxidative stress and aging. Race/ethnic differences in telomere length have been infrequently investigated. Leukocyte telomere length (LTL) was assessed 981 white, black and Hispanic men and women aged 45-84 years participating in the Multi-Ethnic Study of Atherosclerosis. Direct measurement and questionnaire were used to assess covariates. Linear regression was used to estimate associations of LTL with race/ethnicity and age after adjustment for sex, income, education, smoking, physical activity, diet and body mass index. On average blacks and Hispanics had shorter telomeres than whites [adjusted mean differences (standard error) in T/S ratio compared to whites: -0.041 (0.018) for blacks and -0.044 (0.018) for Hispanics]. Blacks and Hispanics showed greater differences in telomere length associated with age than whites (adjusted mean differences in T/S ratio per 1 year increase in age -0.0018, -0.0047 and -0.0055 in whites, blacks and Hispanics respectively). Differences in age associations were more pronounced and only statistically significant in women. Race/ethnic differences in LTL may reflect the cumulative burden of differential exposure to oxidative stress (and its predictors) over the lifecourse. PMID- 19302372 TI - Autophagy and amino acid homeostasis are required for chronological longevity in Saccharomyces cerevisiae. AB - Following cessation of growth, yeast cells remain viable in a nondividing state for a period of time known as the chronological lifespan (CLS). Autophagy is a degradative process responsible for amino acid recycling in response to nitrogen starvation and amino acid limitation. We have investigated the role of autophagy during chronological aging of yeast grown in glucose minimal media containing different supplemental essential and nonessential amino acids. Deletion of ATG1 or ATG7, both of which are required for autophagy, reduced CLS, whereas deletion of ATG11, which is required for selective targeting of cellular components to the vacuole for degradation, did not reduce CLS. The nonessential amino acids isoleucine and valine, and the essential amino acid leucine, extended CLS in autophagy-deficient as well as autophagy-competent yeast. This extension was suppressed by constitutive expression of GCN4, which encodes a transcriptional regulator of general amino acid control (GAAC). Consistent with this, GCN4 expression was reduced by isoleucine and valine. Furthermore, elimination of the leucine requirement extended CLS and prevented the effects of constitutive expression of GCN4. Interestingly, deletion of LEU3, a GAAC target gene encoding a transcriptional regulator of branched side chain amino acid synthesis, dramatically increased CLS in the absence of amino acid supplements. In general, this indicates that activation of GAAC reduces CLS whereas suppression of GAAC extends CLS in minimal medium. These findings demonstrate important roles for autophagy and amino acid homeostasis in determining CLS in yeast. PMID- 19302373 TI - Effects of dietary restriction on mortality and age-related phenotypes in the short-lived fish Nothobranchius furzeri. AB - The short-lived annual fish Nothobranchius furzeri shows extremely short captive life span and accelerated expression of age markers, making it an interesting model system to investigate the effects of experimental manipulations on longevity and age-related pathologies. Here, we tested the effects of dietary restriction (DR) on mortality and age-related markers in N. furzeri. DR was induced by every other day feeding and the treatment was performed both in an inbred laboratory line and a longer-lived wild-derived line. In the inbred laboratory line, DR reduced age-related risk and prolonged maximum life span. In the wild-derived line, DR induced early mortality, did not reduce general age related risk and caused a small but significant extension of maximum life span. Analysis of age-dependent mortality revealed that DR reduced demographic rate of aging, but increased baseline mortality in the wild-derived strain. In both inbred- and wild-derived lines, DR prevented the expression of the age markers lipofuscin in the liver and Fluoro-Jade B (neurodegeneration) in the brain. DR also improved performance in a learning test based on conditioning (active avoidance in a shuttle box). Finally, DR induced a paradoxical up-regulation of glial fibrillary acidic protein in the brain. PMID- 19302374 TI - The core-aldehyde 9-oxononanoyl cholesterol increases the level of transforming growth factor beta1-specific receptors on promonocytic U937 cell membranes. AB - Among the broad variety of compounds generated via oxidative reactions in low density lipoproteins (LDL) and subsequently found in the atherosclerotic plaque are aldehydes that are still esterified to the parent lipid, termed core aldehydes. The most represented cholesterol core aldehyde in LDL is 9-oxononanoyl cholesterol (9-ONC), an oxidation product of cholesteryl linoleate. 9-ONC, at a concentration detectable in biological material, markedly up-regulates mRNA expression and protein level of both the pro-fibrogenic and pro-apoptotic cytokine transforming growth factor beta1 (TGF-beta1) and the TGF-beta receptor type I (TbetaRI) in human U937 promonocytic cells. We also observed increased membrane presentation of TGF-beta receptor type II (TbetaRII). Experiments employing the TbetaRI inhibitor SB431542, or the TGFbeta antagonist DANFc chimera, have shown that the effect on TbetaRI is directly induced by 9-ONC, while TbetaRII up-regulation seems stimulated by its specific ligand, i.e. TGFbeta1, over-secreted meanwhile by treated cells. Increased levels of the cytokine and of its specific receptors in 9-ONC-treated cells clearly occurs through stimulation of extracellular signal-regulated kinase 1 and 2 (ERK1/2), as demonstrated by ERK1/2 knockdown experiments using mitogen-activated protein kinase/extracellular signal-regulated kinase 1 and 2 (MEK1 and MEK2) siRNAs, or PD98059, a selective MEK1/2 inhibitor. 9-ONC might thus sustain further vascular remodeling due to atherosclerosis, not simply by stimulating synthesis of the pro fibrogenic cytokine TGF-beta1 in vascular cells, but also and chiefly by enhancing the TGF-beta1 autocrine loop, because of the marked up-regulation of the cytokine's specific receptors TbetaRI and TbetaRII. PMID- 19302375 TI - Nicotinamide phosphoribosyltransferase imparts human endothelial cells with extended replicative lifespan and enhanced angiogenic capacity in a high glucose environment. AB - Endothelial dysfunction is a characteristic of aging-related vascular disease and is worsened during diabetes. High glucose can impair endothelial cell (EC) function through cellular accumulation of reactive oxygen species, an insult that can also limit replicative lifespan. Nicotinamide phosphoribosyltransferase (Nampt), also known as PBEF and visfatin, is rate-limiting for NAD+ salvage from nicotinamide and confers resistance to oxidative stress via SIRT1. We therefore sought to determine if Nampt expression could resist the detrimental effects of high glucose and confer a survival advantage to human vascular EC in this pathologic environment. Human aortic EC were infected with retrovirus encoding eGFP or eGFP-Nampt, and FACS-selected to yield populations with similar, modest transgene expression. Using a chronic glucose exposure model we tracked EC populations to senescence, assessed cellular metabolism, and determined in vitro angiogenic function. Overexpression of Nampt increased proliferation and extended replicative lifespan, and did so preferentially during glucose overload. Nampt expression delayed markers of senescence and limited reactive oxygen species accumulation in high glucose through a modest increase in aerobic glycolysis. Furthermore, tube networks formed by Nampt-overexpressing EC were more extensive and glucose-resistant, in accordance with SIRT1-mediated repression of the anti angiogenic transcription factor, FoxO1. We conclude that Nampt enables proliferating human EC to resist the oxidative stress of aging and of high glucose, and to productively use excess glucose to support replicative longevity and angiogenic activity. Enhancing endothelial Nampt activity may thus be beneficial in scenarios requiring EC-based vascular repair and regeneration during aging and hyperglycemia, such as atherosclerosis and diabetes-related vascular disease. PMID- 19302376 TI - Living with major depression: experiences from families' perspectives. AB - The aim of this study was to describe the ways of living with major depression in families. Seven families with an adult member who suffered major depression participated, in all 18 participants. Data were collected from seven narrative group interviews, one with each family. A qualitative thematic content analysis was used and the text was coded according to its content and further interpreted into themes. Findings consist of five themes: 'Being forced to relinquish control of everyday life'; during depression the family members lost their energy and could not manage everyday life. 'Uncertainty and instability are affecting life'; everyday life in the families was unstable and emotions influenced the atmosphere. 'Living on the edge of the community': the families periodically lived in seclusion. 'Everyday life becomes hard'; everyday life was demanding for everyone in the family and the responsibility shifted between family members. 'Despite everything a way out can be found'; the families as a unit as well as individually had their own ways of coping and finding some kind of satisfaction within the bounds of possibility. The families' experiences were demanding. Children were aware of their parent's depression and were involved in managing everyday life and emotionally affected by the situation. This legitimates the need for nurses and other healthcare professionals to have a family's perspective on the whole situation and to include partners as well as children and take their experiences seriously when supporting and guiding a family. It is important to develop strategies which in particular include children in the planning, provision and receiving of care as well as unburden them with the responsibility. It is also important to support the family in their own coping strategies with a major depressive episode. PMID- 19302377 TI - Preventive psychosocietal support groups: parents' criteria for good quality. AB - The aim of this study is to produce concepts to describe criteria for good quality of support groups for families with children. The research questions were, what is a good support group like according to parents' experiences and what are the criteria for the good quality from the perspective of families? The study sample consisted of 13 small groups. The study participants consisted of 63 mothers and 14 fathers, who accounted for 63% of the small group attendance. Data were collected in Finland through 13 group interviews. Qualitative content analysis of manifest and latent content was used as the method of analysis. Parents appreciated the availability of information, the ability of the parents to discuss matters, the activities, interaction and good group supervision. As the parental views of the characteristics of a good support group are concrete, they can be easily taken into account with implementing family-oriented support group programmes. The concepts of the present study can be used in the future to construct instruments to evaluate quality of support groups from the perspective of families. Further studies should pay more attention to the experiences of drop out families and families that are not satisfied with group services. PMID- 19302378 TI - Sex determination from hand dimensions of North and South Indians. AB - When dismembered human remains are encountered, the decedent's sex may not be obvious. For these scenarios, techniques for sex determination may play a vital part of the medicolegal investigation. Five hundred hands (230 males, 270 females) from North and South Indian region were studied to investigate sexual dimorphism in hand dimensions (hand length, hand breadth, and palm length). The hand index (hand breadth/hand length x 100) and the palm index (hand breadth/palm length x 100) were derived. The hand dimensions show a higher accuracy in sex determination when compared to indices. Of all hand dimensions, hand breadth has the highest accuracy of sex determination in the study group. The sex differences were found to be statistically significant only for the hand index on the left side. The morphometric parameters of the hand show considerable sexual dimorphism in the Indian population while the hand and palm index remain poor sex indicators. The study thus has medicolegal implications when a dismembered hand is brought for examination. PMID- 19302379 TI - Individual camera identification using correlation of fixed pattern noise in image sensors. AB - This paper presents results of experiments related to individual video camera identification using a correlation coefficient of fixed pattern noise (FPN) in image sensors. Five color charge-coupled device (CCD) modules of the same brand were examined. Images were captured using a 12-bit monochrome video capture board and stored in a personal computer. For each module, 100 frames were captured. They were integrated to obtain FPN. The results show that a specific CCD module was distinguished among the five modules by analyzing the normalized correlation coefficient. The temporal change of the correlation coefficient during several days had only a negligible effect on identifying the modules. Furthermore, a positive relation was found between the correlation coefficient of the same modules and the number of frames that were used for image integration. Consequently, precise individual camera identification is enhanced by acquisition of as many frames as possible. PMID- 19302380 TI - The recovery of latent fingermarks from evidence exposed to ionizing radiation*. AB - Continual reports of illicit trafficking incidents involving radioactive materials have prompted authorities to consider the likelihood of forensic evidence being exposed to radiation. In this study, we investigated the ability to recover latent fingermark evidence from a variety of substrates that were exposed to ionizing radiation. Fingermarks deposited on common surfaces, including aluminum, glass, office paper, and plastic, were exposed to doses ranging from 1 to 1000 kGy, in an effort to simulate realistic situations where evidence is exposed to significant doses of radiation from sources used in a criminal act. The fingermarks were processed using routine fingermark detection techniques. With the exception of glass and aluminum substrates, radiolysis had a considerable effect on the quality of the developed fingermarks. The damage to ridge characteristics can, in part, be attributed to chemical interactions between the substrate and the components of the fingermark secretions that react with the detection reagents. PMID- 19302381 TI - Evaluation of the reliability of DNA typing in the process of identification of war victims in Croatia. AB - Aiming to estimate the frequency of various types of errors that can occur in the large-scale process of identification, we identified and compared genotypes of 911 parent-child pairs in the database of 3498 relatives of people that disappeared during the 1991/1992 war in Croatia. Genotypes of 891 pairs (97.8%) were matching, while 20 pairs did not match in one or more loci. Reanalysis of these samples revealed that out of 1822 analyzed genotypes, one genotype was completely wrong, and two genotypes had one wrong allele because of human errors. Five genotypes had a single wrong allele due to either polymerase chain reaction or electrophoresis errors. In five genotypes mutations were the cause of mismatch. Genetic inconsistencies with parentage were found in four "fathers" (4.2%) and three "mothers" (0.36%). As the majority of observed single-locus errors were caused by nonhuman errors, all databases produced with similar technology would probably have comparable level of errors. PMID- 19302382 TI - A Cannabis sativa STR genotype database for Australian seizures: forensic applications and limitations. AB - A genetic database was established with the aim of documenting the genetic diversity of Cannabis sativa in Australia for future utilization in forensic investigations. The database consisted of genotypes at 10 validated short tandem repeat loci for 510 plants representing drug seizures from across Australia and 57 fiber samples. A total of 106 alleles and 314 different genotypes were detected. All fiber samples exhibited unique genotypes while 55% of the drug samples shared a genotype with one or more samples. Shared genotypes were mostly found within seizures; however, some genotypes were found among seizures. Statistical analysis indicated that genotype sharing was a consequence of clonal propagation rather than a lack of genetic resolution. Thus, the finding of shared genotypes among seizures is likely due to either a common supplier, or direct links among seizures. Notwithstanding the potential intelligence information provided by genetic analysis of C. sativa, our database analysis also reveals some present limitations. PMID- 19302383 TI - Developmental validation of the PrepFiler Forensic DNA Extraction Kit for extraction of genomic DNA from biological samples. AB - The PrepFiler Forensic DNA Extraction Kit enables isolation of genomic DNA from a variety of biological samples. The kit facilitates reversible binding of DNA with magnetic particles resulting in high DNA recovery from samples with very low and high quantities of biological materials: 0.1 and 40 microL of human blood (donor 2) provided 14 and 2883 ng of DNA, respectively. Following the revised SWGDAM guidelines, performance of the developed method was investigated using different sample types including saliva on swabs, semen stains on cotton fabric, samples exposed to environment, samples with polymerase chain reaction (PCR) inhibitors, blood stains (on denim, cotton cloth, and FTA paper), and touch evidence-type samples. DNA yields for all samples tested were equal or better than those obtained by both phenol-chloroform extraction and commercial kits tested. DNA obtained from these samples was free of detectable PCR inhibitors. Short tandem repeat profiles were complete, conclusive, and devoid of PCR artifacts. PMID- 19302384 TI - Modification methods of blank pistols in Turkey in 2006. AB - This study describes the modification methods of blank cartridge firing pistols in Turkey. We have examined cases submitted to the Aydin Regional Criminal Laboratory of Turkey in 2006. In total, 95 modified pistols and 300 modified cartridges were examined. The blank cartridge firing pistols are guns which look similar to "real" pistols, however, there are blockages in their barrel in order to prevent the discharge of a bullet. However, as a result of simple modifications, these pistols can be easily converted into "real" firearms. Studied modification methods are removing the obstruction from the barrel, removing the partial obstruction from the barrel, sleeving a smaller diameter tube into the original barrel, using a replacement barrel, and rifling the original barrel. Special cartridges for these modified pistols are also produced. These modified pistols and cartridges were evaluated in respect of the converting methods. PMID- 19302385 TI - Vitreous fluid and/or urine glucose concentrations in 1335 civil aviation accident pilot fatalities. AB - During aviation accident investigations, vitreous fluid and urine samples from pilot fatalities are analyzed for glucose and blood for hemoglobin A(1c) (HbA(1c)) to monitor diabetic pilots and to discover other pilots with undiagnosed/unreported diabetes. The prevalence of elevated glucose concentrations in fatally injured pilots was evaluated by searching the Civil Aerospace Medical Institute's Toxicology Database for the period 1998-2005. Out of 1335 pilots involving 363 vitreous fluid, 365 urine, and 607 vitreous fluid and urine analyses, 43 pilots had elevated glucose in vitreous fluid (>125 mg/dL) and/or in urine (>100 mg/dL). Of the 20 pilots whose blood samples were analyzed, nine had >6% HbA(1c)--four were known diabetics, and five were unknown diabetics. Urinary glucose levels were elevated in all 13 known hyperglycemic pilots. A considerable number of pilots (30 of 43) had elevated glucose and HbA(1c) (5 of 20), suggesting undiagnosed/unreported diabetic conditions. PMID- 19302386 TI - The use of radio-frequency identification tags for labeling dentures--scanning properties. AB - The inclusion of radio-frequency identification (RFID)-tags within dental prostheses has been suggested as means of effectively labeling such devices and permitting rapid and reliable identification of the wearer. Previous studies have suggested that patients will accept denture labeling and recognize the need for such systems. However, they demand systems that are aesthetic, durable, and secure. One concern over the use of RFID-tags is that they could be scanned by third parties without the patient's knowledge. This study categorizes the scanning patterns of RFID-tags both in vitro and in vivo to provide data for patients for the consent process and for forensic dentists to ensure that they are scanning prostheses optimally. The results demonstrate that the RFID chips can only be read when the interrogator is in close proximity to the denture and thus should alleviate any concerns over privacy issues. However, evidence obtained from both the literature and experiments suggests that authorities must agree upon a unified standard for chip and reader specifications and protocols in order to avoid cases in which RFID-tags may fail to be read by an incompatible reader. PMID- 19302387 TI - Identification of prostate-specific antigen and spermatozoa from a mixture of semen and simulated gastric juice. AB - The detection of prostate-specific antigen (PSA) and visualization of spermatozoa from forensic-type samples containing semen exposed to simulated gastric juice was investigated as a support for forensic practice. Samples of simulated gastric juice mixed with semen were prepared and incubated for up to 4 h at 37 degrees C. Samples were deposited on cotton cloth and on ceramic plates and allowed to dry. The samples were examined for the presence of PSA using the Seratec PSA Semiquant immunochromatographic membrane test. Microscope slides were prepared, stained, and analyzed for spermatozoa. Spermatozoa were detected in all samples, and PSA was detected on neat samples and on samples from the ceramic plate after incubation for up to 4 h. PSA was not detected in the samples deposited on cotton cloth at incubation times greater than 15 min. This may serve as a support for examinations performed when vomit or vomit-stained evidence is submitted for analysis. PMID- 19302388 TI - Validation of SRY marker for forensic casework analysis. AB - Determining the gender of the source of forensic DNA evidence is based on the amelogenin test. However, at times the assay may not be indicative of gender assignment, because of deletions at the amelogenin site. Previously, we described successful coamplification of a marker residing within the SRY gene with the short tandem repeat markers from two commercially available human identification kits. The study herein addresses the validation of primers for the target SRY gene regarding specificity, sensitivity, and robustness. Among 115 unrelated male Slovenians no null allele was observed. Repeatable and reliable results were obtained from as little as 25 pg of template DNA, indicating a high sensitivity of detection for the assay. No polymerase chain reaction product was observed even at a concentration of 10 ng/microL of template female DNA. Additionally, the male specific marker could be detected in mixed male and female samples down to a ratio of 1:16. PMID- 19302389 TI - Five-year results of fixed implant-supported rehabilitations with distal cantilevers for the edentulous mandible. AB - OBJECTIVES: The purpose of this study was to evaluate the survival rate, success rate and primary complications associated with mandibular fixed implant-supported rehabilitations with distal cantilevers over 5 years of function. MATERIAL AND METHODS: In this prospective multi-center trial, 45 fully edentulous patients were treated with implant-supported mandibular hybrid prostheses with distal extension cantilevers. Data were collected at numerous time points, including but not limited to: implant placement, abutment placement, final prosthesis delivery, 3 months and 5 years post-loading. Biological, implant and prosthetic parameters defining survival and success were evaluated for each implant including: sulcus bleeding ndex (SBI) at four sites per implant, width of facial and lingual keratinized gingiva (mm), peri-implant mucosal level (mid-facial from the top of the implant collar, measured in mm), modified plaque index (MPI) at four sites per implant, mobility and peri-implant radiolucency. Survival was defined as implants or prostheses that did not need to be replaced. Success rate was defined as meeting well-established criteria that were chosen to indicate healthy peri implant mucosa osseointegration, prostheses success and complications. RESULTS: A total of 237 implants in 45 completely edentulous patients were included in the study. In each patient, four to six implants were placed to support hybrid prostheses with distal cantilevers. Cantilevers ranged in length from 6 to 21 mm, with an average length of 15.6 mm. The ages of the patients ranged from 34 to 78 with a mean age of 59.5 years. The survival rate of implants was 100% (237/237) and for prostheses 95.5% (43/45). The overall treatment success rate was calculated as 86.7% (39/45). Of the six patients that have not met the criteria for success, two patients required replacement of the entire prosthesis and four patients presented >four complications events. CONCLUSION: Fixed implant supported rehabilitation with distal cantilever resulted in a reliable treatment modality over the 5-year observation period. Although biological parameters of MPI, SBI, keratinized tissue and peri-implant mucosal levels showed statistically significant differences over time, the mean values for each patient remained within the normal limits of oral health. Complications were categorized as biological or technical. The majority of complications were technical complications (54/79) and of these most involved fracture of the acrylic teeth and base (20/54). While the survival rate was 100% for implants and 95.5% for prostheses, the application of strict criteria for treatment success resulted in an overall treatment success rate of 86.7%. PMID- 19302390 TI - Osteotome sinus floor elevation technique without grafting material: 3-year results of a prospective pilot study. AB - OBJECTIVE: In a prospective pilot study, short< or =10 mm ITI-SLA implants were placed in the resorbed posterior maxilla by means of an osteotome sinus floor elevation (OSFE) procedure without grafting material. This paper presents 3-year data assessing bone-level changes around implants. MATERIAL AND METHODS: Twenty five implants were placed in 17 patients to rehabilitate 16 molar and nine premolar sites. The mean residual bone height (RBH) was 5.4 +/- 2.3 mm. A healing period of 3-4 months was allowed before abutment tightening. Most implants (21/25) were 10 mm long, and the others were 8 and 6 mm long. At the 3-year control, endo-sinus bone gain, implant length protruding into the sinus and crestal bone loss (CBL) were measured on periapical radiographs. RESULTS: All implants fulfilled the survival criteria. Despite the absence of grafting material, implants were embedded into newly formed bone tissue. All implants gained endo-sinus bone; the mean gained bone was 3.1 +/- 1.5 mm. The residual protrusion length decreased from 4.9 +/- 2.1 to 1.8 +/- 2.1 mm. CBL was 0.9 +/- 0.8 mm. CONCLUSIONS: This study confirms that the OSFE procedure without grafting material is sufficient to create bone beyond the natural limit of the sinus. On the mid-term of 3 years, the technique was found to be predictable in the posterior maxilla when the RBH is limited. Implants gained endo-sinus bone despite the lack of grafting material. Bone gain was still improving over the first-year control. No shrinkage of the augmented area was observed. PMID- 19302391 TI - Histometric evaluation of bone around titanium implants with different surface treatments in rats exposed to cigarette smoke inhalation. AB - OBJECTIVES: There is a lack of histological information about the influence of cigarette smoke on bone around surface-treated implants. The aim of the present study was to test the influence of titanium surface treatment on osseointegration in animals that were exposed to intermittent cigarette smoke inhalation. MATERIAL AND METHODS: Twenty-two male Wistar rats were used. One tibia, chosen at random, received a machined titanium implant (MI) while the other received an aluminum oxide-blasted surface implant (ABI). The animals were randomly assigned to one of the following groups: Group 1 - control (n=11) and Group 2 - intermittent cigarette smoke inhalation (n=11). Sixty days after surgery, the animals were sacrificed. The degree of bone-to-implant contact (BIC), bone filling (BF) within the limits of the threads of the implants and bone density (proportion of mineralized bone in a 500-mum-wide zone lateral to the implant - BD) were measured in the cortical (zone A) and cancellous bone (zone B) areas. RESULTS: Data analysis showed significant differences when comparing the groups and implant surfaces in both zones for BIC (two-way ANOVA -P<0.05). The two groups presented higher BIC mean values for ABI, when compared with MI (P<0.05). In group 2, cigarette smoke inhalation negatively affected BF in both zones (P<0.05). Group 2 presented a significantly decreased BD in both zones (P<0.05). No statistically significant differences were observed between surfaces in any of the groups for BD. CONCLUSION: Within the limits of the present study, it can be concluded that the aluminum oxide blast surface treatment may increase the degree of BIC but cannot overcome the detrimental effect of tobacco smoke on bone around titanium implants. PMID- 19302392 TI - Stability change of chemically modified sandblasted/acid-etched titanium palatal implants. A randomized-controlled clinical trial. AB - AIM: The aim of this randomized-controlled clinical study was to examine stability changes of palatal implants with chemically modified sandblasted/acid etched (modSLA) titanium surface compared with a standard SLA surface, during the early stages of bone healing. MATERIALS AND METHODS: Forty adult volunteers were recruited and randomly assigned to the test group (modSLA surface) and to the control group (SLA surface). The test and control implants had the same microscopic and macroscopic topography, but differed in surface chemistry. To document implant stability changes resonance frequency analysis (RFA) was performed at implant insertion, at 7, 14, 21, 28, 35, 42, 49, 56, 70 and 84 days thereafter. RFA values were expressed as an implant stability quotient (ISQ). RESULTS: Immediately after implant installation, the ISQ values for both surfaces tested were not significantly different and yielded mean values of 73.8+/-5 for the control and 72.7+/-3.9 for the test surface. In the first 2 weeks after implant installation, both groups showed only small changes and thereafter a decreasing trend in the mean ISQ levels. In the test group, after 28 days a tendency towards increasing ISQ values was observed and 42 days after surgery the ISQ values corresponded to those after implant insertion. For the SLA-control group, the trend changed after 35 days and yielded ISQ values corresponding to the baseline after 63 days. After 12 weeks of observation, the test surface yielded significantly higher stability values of 77.8+/-1.9 compared with the control implants of 74.5+/-3.9, respectively. CONCLUSION: The results support the potential for chemical modification of the SLA surface to positively influence the biologic process of osseointegration and to decrease the healing time. PMID- 19302393 TI - Use of a new cross-linked collagen membrane for the treatment of dehiscence-type defects at titanium implants: a prospective, randomized-controlled double-blinded clinical multicenter study. AB - OBJECTIVES: The aim of the present randomized-controlled double-blinded clinical multicenter study was to assess the use of either a new cross-linked (VN) or a native collagen membrane (BG) for the treatment of dehiscence-type defects at titanium implants. MATERIAL AND METHODS: A total of n=54 patients were recruited in four German university clinics. According to a parallel-groups design, dehiscence-type defects at titanium implants were filled with a natural bone mineral and randomly assigned to either VN or BG. Submerged sites were allowed to heal for 4 months. Primary (e.g., changes in defect length - DeltaDL, quality of newly formed tissue [0-4] - TQ) and secondary parameters (e.g., membrane exposure, tissue conditions at dehisced sites) were consecutively recorded. RESULTS: Four patients were excluded due to an early wound infection (VN:3; BG:1), and one patient was lost during follow-up (VN). The mean DeltaDL was 3.0 +/- 2.5 mm in the VN, and 1.94 +/- 2.13 mm in the BG group. The assessment of TQ revealed comparable mean values in both groups (VN: 3.05 +/- 1.66, BG: 3.46 +/- 1.48). A significant correlation between membrane exposure and inflammation of the adjacent soft tissue was observed in the VN group. In both groups, the mean DL and TQ values were not significantly different at either non-exposed or exposed implant sites. CONCLUSION: The results of the present study have indicated that VN supported bone regeneration on a level non-inferior to BG. However, in case of a premature membrane exposure, cross-linking might impair soft-tissue healing or may even cause wound infections. PMID- 19302394 TI - Differential cytokine expressions affect the severity of peri-implant disease. AB - OBJECTIVE: This study assessed gene expression by quantitative polymerase chain reaction of inflammatory- [interleukin (IL)-12, tumor necrosis factor-alpha (TNF alpha), IL-4, and IL-10] and osteoclastogenesis-related factors [receptor activator of NF-kappaB ligand (RANKL) and osteoprotegerin (OPG)] in sites exhibiting different severities of peri-implant disease. MATERIAL AND METHODS: Peri-implant soft tissue biopsies (n=48) were harvested from healthy implant (HI), mucositis (MC), initial peri-implantitis (IP) and severe peri-implantitis (SP) sites. RESULTS: IL-12 and TNF-alpha mRNA levels were higher in SP, followed by IP and MC (P <0.05). IL-4 was higher in HI, followed by MC, SP and IP (P <0.05). IL-10 was the lowest in HI, while no differences were detected among the diseased groups (P>0.05). OPG mRNA levels were higher in HI, followed by IP, SP and MC, whereas RANKL was increased as the peri-implantitis severity increased (P<0.05). The highest OPG/RANKL ratio was observed in HI and the lowest in SP (P<0.01). CONCLUSION: These findings suggest that expressions of inflammatory- and osteoclastogenesis-related factors may play an important role in the onset and severity of the peri-implant diseases. PMID- 19302395 TI - Vertical bone augmentation with fluvastatin in an injectable delivery system: a rat study. AB - OBJECTIVES: HMG-CoA reductase inhibitors (statins) are widely used for hyperlipidemia. Previous studies demonstrate that statins stimulate bone morphogenetic protein-2 (BMP-2) expression and lead to bone formation. The aim of this study was to evaluate whether percutaneously injected statin with a novel statin delivery system achieved vertical bone augmentation. MATERIAL AND METHODS: As experimental groups, atelocollagen-alpha-tricalcium phosphate (alphaTCP) composites containing 3.3 mg (low dose) or 6.7 mg (high dose) of fluvastatin were injected (one shot) subcutaneously over the calvarial periosteum of rats. The animals were then sacrificed 1, 2, and 4 weeks after injection. Vertically augmented bone was assessed by histomorphometric procedures, i.e., by measuring new bone thickness (NBT) and bone density (BD). RESULTS: In control groups, no newly formed bone could be seen over the calvarial bone. In the experimental groups, in contrast, a large amount of newly formed bone could be seen over the preexisting calvarial bone. The newly formed bone was seen to be in direct contact with the preexisting bone. During the entire observation, significant NBT was observed in the experimental groups (P<0.05). At the final stage of observation (4 weeks), NBT was 66.7% (low-dose group) and 59.7% (high-dose group), while they were from 1% to 16.3% in the control groups. In the experimental groups, BD significantly increased in a time-dependent manner. CONCLUSION: Percutaneously applied fluvastatin (one shot) with a composite of alphaTCP and collagen has great potential to augment the height of the bone. PMID- 19302396 TI - Linking movement behaviour, dispersal and population processes: is individual variation a key? AB - 1. Movement behaviour has become increasingly important in dispersal ecology and dispersal is central to the development of spatially explicit population ecology. The ways in which the elements have been brought together are reviewed with particular emphasis on dispersal distance distributions and the value of mechanistic models. 2. There is a continuous range of movement behaviours and in some species, dispersal is a clearly delineated event but not in others. The biological complexities restrict conclusions to high-level generalizations but there may be principles that are common to dispersal and other movements. 3. Random walk and diffusion models when appropriately elaborated can provide an understanding of dispersal distance relationships on spatial and temporal scales relevant to dispersal. Leptokurtosis in the relationships may be the result of a combination of factors including population heterogeneity, correlation, landscape features, time integration and density dependence. The inclusion in diffusion models of individual variation appears to be a useful elaboration. The limitations of the negative exponential and other phenomenological models are discussed. 4. The dynamics of metapopulation models are sensitive to what appears to be small differences in the assumptions about dispersal. In order to represent dispersal realistically in population models, it is suggested that phenomenological models should be replaced by those based on movement behaviour incorporating individual variation. 5. The conclusions are presented as a set of candidate principles for evaluation. The main features of the principles are that uncorrelated or correlated random walk, not linear movement, is expected where the directions of habitat patches are unpredictable and more complex behaviour when organisms have the ability to orientate or navigate. Individuals within populations vary in their movement behaviour and dispersal; part of this variation is a product of random elements in movement behaviour and some of it is heritable. Local and metapopulation dynamics are influenced by population heterogeneity in dispersal characteristics and heritable changes in dispersal propensity occur on time-scales short enough to impact population dynamics. PMID- 19302397 TI - Perceived health status as measured by the SF-36 in patients with multiple sclerosis: a review. AB - This review of literature gives an overview of recent studies about perceived health status as measured by the Short-Form-36 (SF-36) Health Survey in patients with multiple sclerosis (MS). The SF-36 is one of the tools measuring health status in patients used in international research and clinical practice. It measures two main health concepts - physical and mental. The SF-36 represents a valid instrument able to detect differences in perceived health status in patients. A computer-aided search in Medline and PsycINFO resulted in 504 articles in English published from 1996 to August 2006. After the screening process on the basis of abstracts, eight articles consisting of empirical studies remained in which perceived health status was evaluated using the SF-36 Health Survey. Seven studies focused on disability and perceived health status in the SF 36. Two studies focused on the relationship between depression and perceived health status. These studies showed that MS patients with low disability and minor depression scored significantly better than patients with high disability and major depression in the SF-36 health dimensions. Gender seems to have no influence on perceived health status in MS patients. The longer the disease duration and the more severe the disease, the lower the patients scored in perceived health status. The more disabled, the more depressive and the older the patients, the poorer their perceived health status was. Health providers supporting appropriate treatment might pay more attention to more disabled and more depressive patients, with longer disease duration. Perceived health status can be a predictor of prognosis and intervention outcomes. The study shows the importance of measuring perceived health status in MS patients with implications for their quality of life and provision of care. PMID- 19302398 TI - Study on the impact of caregivers in an Italian high specialization hospital: presence, costs and nurse's perception. AB - The study analysed two key questions: (i) the prevalence of informal caregiving in medical and surgical wards of a high specialization hospital; (ii) the reasonable cost for the structure that would have to pay to replace informal caregiving? The study was conducted in June 2006 as a prevalence survey, using a questionnaire administered to informal caregivers and nurses working in medical and surgical wards of a high specialization hospital by ad hoc trained personnel. Questionnaire consisted in three sections: the first focused on patient's characteristics, the second on caregivers and the third on nurses' perception on caregiving phenomenon. One hundred and twenty-four eligible caregivers were identified. During the study patients admitted to hospital medical and surgery wards were 520. Among these 16.5% (86/520) was assisted by one or more caregivers. Caregivers' response rate was 69.4% (86/124), corresponding to 66 patients. This study yielded an average of 455.9 minutes per day (SD = 370.2; range = 120-1440) or 52.9 hours per week. Caregiver's presence was recognized in 88.9% (56/63) of patients. Despite the societal perspective, the costs and effects of informal caregiving to the informal caregiver are often ignored in economic evaluation. The costs of informal care are an important extent related to time inputs by relatives and friends of the care recipients. Our approach has been to monetize the informal activity care contribution of family members and/or caregivers. PMID- 19302399 TI - Validation of the direct analysis in real time source for use in forensic drug screening. AB - The Direct Analysis in Real Time (DART) ion source is a relatively new mass spectrometry technique that is seeing widespread use in chemical analyses world wide. DART studies include such diverse topics as analysis of flavors and fragrances, melamine in contaminated dog food, differentiation of writing inks, characterization of solid counterfeit drugs, and as a detector for planar chromatography. Validation of this new technique for the rapid screening of forensic evidence for drugs of abuse, utilizing the DART source coupled to an accurate mass time-of-flight mass spectrometer, was conducted. The study consisted of the determination of the lower limit of detection for the method, determination of selectivity and a comparison of this technique to established analytical protocols. Examples of DART spectra are included. The results of this study have allowed the Virginia Department of Forensic Science to incorporate this new technique into their analysis scheme for the screening of solid dosage forms of drugs of abuse. PMID- 19302400 TI - Trace elemental analysis of titanium dioxide pigments and automotive white paint fragments for forensic examination using high-energy synchrotron radiation x-ray fluorescence spectrometry. AB - High-energy synchrotron radiation x-ray fluorescence spectrometry (SR-XRF) utilizing 116 keV x-rays was used to characterize titanium dioxide pigments (rutile) and automotive white paint fragments for forensic examination. The technique allowed analysis of K lines of 9 trace elements in 18 titanium dioxide pigments (rutile), and 10 trace elements in finish coat layers of seven automotive white paint fragments. High-field strength elements (HFSE) were found to strongly reflect the origin of the titanium dioxide (TiO(2)) pigments, and could be used as effective parameters for discrimination and classification of the pigments and paint fragments. A pairwise comparison of the finish coat layers of seven automotive white paint fragments was performed. The trace elements in the finish coat layers detected by the high-energy SR-XRF were especially effective for identification. By introducing the trace element information of primer and electrocoat layers, all the automotive white paint fragments could be discriminated by this technique. PMID- 19302401 TI - Microscopic age estimation from the anterior cortex of the femur in Korean adults. AB - The purpose of this study was to develop age-predicting equations from the anterior cortex of the femur of Korean adults. Seventy-two femoral samples (44 male and 28 female) were obtained from Korean cadavers and used to develop the equations. The thin sections (<100-microm thick) were prepared by manual grinding; the sections were not decalcified and were stained with Villanueva bone stain reagent. Analysis of covariance showed no significant differences in age adjusted histomorphological variables between sexes. In stepwise regression analysis, osteon population density, average osteon area, and the most anterior cortical width were selected for an age-predicting equation which produced a high regression correlation (R(2) = 0.789). The average Haversian canal area was not significantly related to age for any specimen. PMID- 19302402 TI - Vicious dogs: the antisocial behaviors and psychological characteristics of owners. AB - Each year, 4.7 million people are bitten by dogs. Of those bitten each year, 386,000 are seriously injured and some killed. Consequently, many insurance companies refuse to issue homeowners insurance to owners of specific breeds of dogs considered "vicious" or high risk of causing injury. This study examined whether vicious dog owners were different on antisocial behaviors and personality dimensions. A total of 869 college students completed an anonymous online questionnaire assessing type of dog owned, criminal behaviors, attitudes towards animal abuse, psychopathy, and personality. The sample was divided into four groups: vicious dog owners, large dog owners, small dog owners, and controls. Findings revealed vicious dog owners reported significantly more criminal behaviors than other dog owners. Vicious dog owners were higher in sensation seeking and primary psychopathy. Study results suggest that vicious dog ownership may be a simple marker of broader social deviance. PMID- 19302403 TI - Mosaic manifestations of monogenic skin diseases. AB - A genetic mosaic is defined as an organism which is composed of genetically different cell lines which originate from a homogeneous zygote. Etiologically, cutaneous mosaics can be divided into two large categories, epigenetic mosaicism and genomic mosaicism. Genomic mosaics which have two or more genetically different cell populations are not inherited with the exception of para-dominant inheritance pattern. Epigenetic mosaics have a structurally homogeneous cell population but there are functional differences induced by modifying factors in the form of gene-steering retroviral elements that can be inherited. We distinguish five different manifestation patterns of mosaicism, including the Blaschko lines pattern, patchy pattern without midline separation, checkerboard pattern, phylloid pattern and lateralization pattern. All forms of epigenetic mosaicism, including the various patterns of X-inactivation, appear to be caused by the action of retrotransposons. A new concept is functional autosomal mosaicism transmittable through the action of retrotransposons PMID- 19302404 TI - Detecting methylation patterns of p16, MGMT, DAPK and E-cadherin genes in multiple myeloma patients. AB - Multiple myeloma (MM) is a B-cell neoplasia characterized by the clonal proliferation of plasma cells. Besides known genetic abnormalities, epigenetic changes are also known to effect MM pathogenesis. DNA methylation is an epigenetic mechanism that silences genes by adding methyl groups to cytosine guanine dinucleotides at the promoter regions. In this study, the methylation status of four genes; p16, O6-methyl guanine DNA methyl transferase (MGMT), death associated protein kinase (DAPK) and E-cadherin (ECAD); at the time of diagnosis was investigated using methylation-specific polymerase chain reaction (MS-PCR). In the 20 cases studied; methylation of the promoter regions of p16, MGMT, DAPK and ECAD genes was detected in 10%, 40%, 10% and 45% of the cases, respectively. In 65% (13/20) of cases, at least one of the genes studied had promoter methylation; while 35% of cases (7/20) had methylated promoters of more than one gene. There was a significant correlation between promoter hypermethylation of MGMT and the presence of extramedullary involvement; but for the other genes no correlation was found regarding disease properties like age, disease stage, clinical course and the presence of lytic bone lesions. Determining the methylation profiles of genes in MM, could lead to a new understanding of the disease pathogenesis and guide the assessment of treatment options. PMID- 19302405 TI - Modeling data with excess zeros and measurement error: application to evaluating relationships between episodically consumed foods and health outcomes. AB - Dietary assessment of episodically consumed foods gives rise to nonnegative data that have excess zeros and measurement error. Tooze et al. (2006, Journal of the American Dietetic Association 106, 1575-1587) describe a general statistical approach (National Cancer Institute method) for modeling such food intakes reported on two or more 24-hour recalls (24HRs) and demonstrate its use to estimate the distribution of the food's usual intake in the general population. In this article, we propose an extension of this method to predict individual usual intake of such foods and to evaluate the relationships of usual intakes with health outcomes. Following the regression calibration approach for measurement error correction, individual usual intake is generally predicted as the conditional mean intake given 24HR-reported intake and other covariates in the health model. One feature of the proposed method is that additional covariates potentially related to usual intake may be used to increase the precision of estimates of usual intake and of diet-health outcome associations. Applying the method to data from the Eating at America's Table Study, we quantify the increased precision obtained from including reported frequency of intake on a food frequency questionnaire (FFQ) as a covariate in the calibration model. We then demonstrate the method in evaluating the linear relationship between log blood mercury levels and fish intake in women by using data from the National Health and Nutrition Examination Survey, and show increased precision when including the FFQ information. Finally, we present simulation results evaluating the performance of the proposed method in this context. PMID- 19302406 TI - Nonparametric modeling of longitudinal covariance structure in functional mapping of quantitative trait loci. AB - Estimation of the covariance structure of longitudinal processes is a fundamental prerequisite for the practical deployment of functional mapping designed to study the genetic regulation and network of quantitative variation in dynamic complex traits. We present a nonparametric approach for estimating the covariance structure of a quantitative trait measured repeatedly at a series of time points. Specifically, we adopt Huang et al.'s (2006, Biometrika 93, 85-98) approach of invoking the modified Cholesky decomposition and converting the problem into modeling a sequence of regressions of responses. A regularized covariance estimator is obtained using a normal penalized likelihood with an L(2) penalty. This approach, embedded within a mixture likelihood framework, leads to enhanced accuracy, precision, and flexibility of functional mapping while preserving its biological relevance. Simulation studies are performed to reveal the statistical properties and advantages of the proposed method. A real example from a mouse genome project is analyzed to illustrate the utilization of the methodology. The new method will provide a useful tool for genome-wide scanning for the existence and distribution of quantitative trait loci underlying a dynamic trait important to agriculture, biology, and health sciences. PMID- 19302407 TI - Modeling age and nest-specific survival using a hierarchical Bayesian approach. AB - Recent studies have shown that grassland birds are declining more rapidly than any other group of terrestrial birds. Current methods of estimating avian age specific nest survival rates require knowing the ages of nests, assuming homogeneous nests in terms of nest survival rates, or treating the hazard function as a piecewise step function. In this article, we propose a Bayesian hierarchical model with nest-specific covariates to estimate age-specific daily survival probabilities without the above requirements. The model provides a smooth estimate of the nest survival curve and identifies the factors that are related to the nest survival. The model can handle irregular visiting schedules and it has the least restrictive assumptions compared to existing methods. Without assuming proportional hazards, we use a multinomial semiparametric logit model to specify a direct relation between age-specific nest failure probability and nest-specific covariates. An intrinsic autoregressive prior is employed for the nest age effect. This nonparametric prior provides a more flexible alternative to the parametric assumptions. The Bayesian computation is efficient because the full conditional posterior distributions either have closed forms or are log concave. We use the method to analyze a Missouri dickcissel dataset and find that (1) nest survival is not homogeneous during the nesting period, and it reaches its lowest at the transition from incubation to nestling; and (2) nest survival is related to grass cover and vegetation height in the study area. PMID- 19302408 TI - Bayesian wombling for spatial point processes. AB - In many applications involving geographically indexed data, interest focuses on identifying regions of rapid change in the spatial surface, or the related problem of the construction or testing of boundaries separating regions with markedly different observed values of the spatial variable. This process is often referred to in the literature as boundary analysis or wombling. Recent developments in hierarchical models for point-referenced (geostatistical) and areal (lattice) data have led to corresponding statistical wombling methods, but there does not appear to be any literature on the subject in the point-process case, where the locations themselves are assumed to be random and likelihood evaluation is notoriously difficult. We extend existing point-level and areal wombling tools to this case, obtaining full posterior inference for multivariate spatial random effects that, when mapped, can help suggest spatial covariates still missing from the model. In the areal case we can also construct wombled maps showing significant boundaries in the fitted intensity surface, while the point-referenced formulation permits testing the significance of a postulated boundary. In the computationally demanding point-referenced case, our algorithm combines Monte Carlo approximants to the likelihood with a predictive process step to reduce the dimension of the problem to a manageable size. We apply these techniques to an analysis of colorectal and prostate cancer data from the northern half of Minnesota, where a key substantive concern is possible similarities in their spatial patterns, and whether they are affected by each patient's distance to facilities likely to offer helpful cancer screening options. PMID- 19302409 TI - Shrinkage-based diagonal discriminant analysis and its applications in high dimensional data. AB - High-dimensional data such as microarrays have brought us new statistical challenges. For example, using a large number of genes to classify samples based on a small number of microarrays remains a difficult problem. Diagonal discriminant analysis, support vector machines, and k-nearest neighbor have been suggested as among the best methods for small sample size situations, but none was found to be superior to others. In this article, we propose an improved diagonal discriminant approach through shrinkage and regularization of the variances. The performance of our new approach along with the existing methods is studied through simulations and applications to real data. These studies show that the proposed shrinkage-based and regularization diagonal discriminant methods have lower misclassification rates than existing methods in many cases. PMID- 19302410 TI - Protective effects of exercise preconditioning on hindlimb unloading-induced atrophy of rat soleus muscle. AB - AIM: A chronic decrease in the activation and loading levels of skeletal muscles as occurs with hindlimb unloading (HU) results in a number of detrimental changes. Several proteolytic pathways are involved with an increase in myofibrillar protein degradation associated with HU. Exercise can be used to counter this increase in proteolytic activity and, thus, may be able to protect against some of the detrimental changes associated with chronic decreased use. The purpose of the present study was to determine the potential of a single bout of preconditioning endurance exercise in attenuating the effects of 2 weeks of HU on the mass, phenotype and force-related properties of the soleus muscle in adult rats. METHODS: Male Wistar rats were subjected to HU for 2 weeks. One half of the rats performed a single bout of treadmill exercise for 25 min immediately prior to the 2 weeks of HU. RESULTS: Soleus mass, maximum tetanic tension, myofibrillar protein content, fatigue resistance and percentage of type I (slow) myosin heavy chain were decreased in HU rats. In addition, markers for the cathepsin, calpain, caspase and ATP-ubiquitin-proteasome proteolytic pathways were increased. The preconditioning endurance exercise bout attenuated all of the detrimental changes associated with HU, and increased HSP72 mRNA expression and protein levels. CONCLUSION: These findings indicate that exercise preconditioning may be an effective countermeasure to the detrimental effects of chronic decreases in activation and loading levels on skeletal muscles and that an elevation in HSP72 may be one of the mechanisms associated with these responses. PMID- 19302412 TI - Riding shotgun: a dual role for the epidermal growth factor-Cripto/FRL-1/Cryptic protein Cripto in Nodal trafficking. AB - Nodal is a secreted protein of the transforming growth factor beta (TGFbeta) family that activates Smad2 and Smad3 transcription factors through complexes of type I and type II activin receptors and glycosyl-phosphatidylinositol-anchored coreceptors of the epidermal growth factor-like Cripto/FRL-1/Cryptic family. During early embryogenesis, it stimulates the proliferation of pluripotent progenitor cells and specifies, in a dosage-dependent manner, their subsequent allocation to distinct germ layers. Available evidence indicates that the signaling strength of Nodal is controlled at the level of endocytic uptake and turnover of activated receptor complexes in early endosomes, but insights into the underlying molecular mechanisms are still limited. In this review, I briefly survey literature on the trafficking of the related TGFbeta receptors, and I discuss recent findings indicating that endocytosis of Nodal is coupled to proteolytic processing of its precursor at the cell surface and that the maturation and internalization of Nodal need to be guided by Cripto to stabilize endosomal signaling platforms. PMID- 19302411 TI - Photoreceptor IFT complexes containing chaperones, guanylyl cyclase 1 and rhodopsin. AB - Intraflagellar transport (IFT) provides a mechanism for the transport of cilium specific proteins, but the mechanisms for linkage of cargo and IFT proteins have not been identified. Using the sensory outer segments (OS) of photoreceptors, which are derived from sensory cilia, we have identified IFT-cargo complexes containing IFT proteins, kinesin 2 family proteins, two photoreceptor-specific membrane proteins, guanylyl cyclase 1 (GC1, Gucy2e) and rhodopsin (RHO), and the chaperones, mammalian relative of DNAJ, DnajB6 (MRJ), and HSC70 (Hspa8). Analysis of these complexes leads to a model in which MRJ through its binding to IFT88 and GC1 plays a critical role in formation or stabilization of the IFT-cargo complexes. Consistent with the function of MRJ in the activation of HSC70 ATPase activity, Mg-ATP enhances the co-IP of GC1, RHO, and MRJ with IFT proteins. Furthermore, RNAi knockdown of MRJ in IMCD3 cells expressing GC1-green fluorescent protein (GFP) reduces cilium membrane targeting of GC1-GFP without apparent effect on cilium elongation. PMID- 19302413 TI - Women's well-being: the role of individual differences. AB - The present study examined the relationship between physical and psychological well-being, personality characteristics and demographic variables related to motherhood, work and marital status in a sample of 3,723 Dutch women. Analysis revealed several interesting relationships. First, whereas neuroticism was negatively related to both physical and psychological well-being, extraversion was positively related to well-being. Second, a clear positive relationship emerged between work participation and both types of well-being. Results are discussed in terms of the vulnerability stress model. PMID- 19302414 TI - Auditory processing in developmental dyslexia: an exploratory study of an auditory and visual matching training program with Swedish children with developmental dyslexia. AB - This study examined whether training using a nonverbal auditory-visual matching task had a remedial effect on reading skills in developmental dyslexia. The pretest/post-test design was used with Swedish children (N= 41), between the ages of 7 and 12. Training comprised twice-weekly sessions of 15 minutes, over eight weeks. There was an improvement in auditory-visual matching during the training period. There were also improvements in some reading test scores, especially in reading nonsense words and in reading speed. These improvements in tasks which are thought to rely on phonological processing suggest that such reading difficulties in dyslexia may stem in part from more basic perceptual difficulties, including those required to manage the visual and auditory components of the decoding task. The utility of the concept of auditory structuring is discussed in relation to auditory and phonological processing skills when a child learns to read. PMID- 19302415 TI - The correlation of hepatocyte nuclear factor 4 alpha and 3 beta with hepatitis B virus replication in the liver of chronic hepatitis B patients. AB - Hepatocyte nuclear factors 4 alpha (HNF4alpha) and 3 beta (HNF3beta) are members of a group of liver-enriched transcription factors (LETFs) that play important roles in regulating the replication of hepatitis B virus (HBV). Using cell culture and animal models, we showed that HNF4alpha supports HBV replication in nonhepatic cells and HNF3beta inhibits HBV replication. However, the expression of HNF4alpha and HNF3beta in the liver tissue of chronic HBV-infected patients and the relationship between the levels of HNF4alpha and HNF3beta and HBV replication are unclear. In this study, liver biopsy specimens from 86 chronic HBV-infected patients were collected. The expression levels of HNF4alpha, HNF3beta, hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) were detected by an immunohistochemical technique and the level of HBV DNA was checked by in situ hybridization with serial sections from liver biopsy tissue samples. We show here that samples with higher levels of HNF4alpha expression also have higher levels of HBsAg, HBcAg and HBV DNA. In contrast, in samples with higher levels of HNF3beta expression, levels of HBsAg, HBcAg and HBV DNA were lower. There was a positive correlation between HNF4alpha expression and HBV replication, and a negative correlation between HNF3beta expression and HBV replication, in the liver of chronic HBV-infected patients. This suggests that HNF4alpha and HNF3beta likely participate in HBV replication in patients with HBV infection, or that HBV replication may somehow influence the expression of HNF4alpha and HNF3beta in the liver. PMID- 19302416 TI - Functional characteristics of apheresis-derived platelets treated with ultraviolet light combined with either amotosalen-HCl (S-59) or riboflavin (vitamin B2) for pathogen-reduction. AB - BACKGROUND: To examine if different pathogen-reduction technologies (PRTs) induce different degrees of platelet (PLT) storage lesion. DESIGN: Twenty-seven split triple-dose apheresis PLTs were PRT treated using ultraviolet light with either riboflavin (M) or psoralen (I) or remained untreated (C). Samples taken on days (d) 0 to 8 were analysed for PLT count, blood gas (pH, pO(2) and pCO(2)), metabolism (lactate, glucose, ATP content), in vitro function [swirling, hypotonic shock response (HSR) and aggregation], activation (p-selectin expression) and cellular integrity (JC-1 signal, annexin A5 release). RESULTS: Platelet counts of all study groups remained unchanged during storage indicating that PRT treatment did not induce relevant cell lysis. Although M units demonstrated the highest values for HSR until d5, PRT treatment lowered all parameters examined with significant differences to untreated controls by d7 of storage. During final storage, M was significantly superior over I for HSR, aggregation with TRAP-6 as agonist (collagen was similar), annexin A5 release and JC-1 signal. Regarding blood gas and metabolic analysis, the most evident effect of PRT was an elevated glycolytic flux combined with higher acidity due to increased lactate accumulation. Most likely due to impaired O(2) consumption, pH and ATP decreased more rapidly in I relative to C and M. CONCLUSION: Pathogen reduction technology-treated PLTs remained comparable to untreated units throughout 7 days of storage. Mitochondria-based oxidative respiration appeared up-regulated after the riboflavin-based PRT. Compared to the psoralen-based PRT, this resulted in significantly better ATP maintenance and in vitro function during the last storage period (d7, d8). PMID- 19302417 TI - Photosynthesis in cells around veins of the C(3) plant Arabidopsis thaliana is important for both the shikimate pathway and leaf senescence as well as contributing to plant fitness. AB - Cells associated with veins of C(3) species often contain significant amounts of chlorophyll, and radiotracer analysis shows that carbon present in the transpiration stream may be used for photosynthesis in these cells. It is not clear whether CO2 is also supplied to these cells close to veins via stomata, nor whether this veinal photosynthesis supplies carbon skeletons to particular metabolic pathways. In addition, it has not been possible to determine whether photosynthesis in cells close to veins of C(3) plants is quantitatively important for growth or fitness. To investigate the role of photosynthesis in cells in and around the veins of C(3) plants, we have trans-activated a hairpin construct to the chlorophyll synthase gene (CS) using an Arabidopsis thaliana enhancer trap line specific to veins. CS is responsible for addition of the phytol chain to the tetrapyrolle head group of chlorophyll, and, as a result of cell-specific trans activation of the hairpin to CS, chlorophyll accumulation is reduced around veins. We use these plants to show that, under steady-state conditions, the extent to which CO2 is supplied to cells close to veins via stomata is limited. Fixation by minor veins of CO2 supplied to the xylem stream and the amount of specific metabolites associated with carbohydrate metabolism and the shikimate pathway were all reduced. In addition, an abundance of transcripts encoding components of pathways that generate phosphoenolpyruvate were altered. Leaf senescence, growth rate and seed size were all reduced in the lines with lower photosynthetic ability in veins and in cells close to veins. PMID- 19302418 TI - Calcium elevation-dependent and attenuated resting calcium-dependent abscisic acid induction of stomatal closure and abscisic acid-induced enhancement of calcium sensitivities of S-type anion and inward-rectifying K channels in Arabidopsis guard cells. AB - Stomatal closure in response to abscisic acid depends on mechanisms that are mediated by intracellular [Ca2+] ([Ca2+]i), and also on mechanisms that are independent of [Ca2+]i in guard cells. In this study, we addressed three important questions with respect to these two predicted pathways in Arabidopsis thaliana. (i) How large is the relative abscisic acid (ABA)-induced stomatal closure response in the [Ca2+]i-elevation-independent pathway? (ii) How do ABA insensitive mutants affect the [Ca2+]i-elevation-independent pathway? (iii) Does ABA enhance (prime) the Ca2+ sensitivity of anion and inward-rectifying K+ channel regulation? We monitored stomatal responses to ABA while experimentally inhibiting [Ca2+]i elevations and clamping [Ca2+]i to resting levels. The absence of [Ca2+]i elevations was confirmed by ratiometric [Ca2+]i imaging experiments. ABA-induced stomatal closure in the absence of [Ca2+]i elevations above the physiological resting [Ca2+]i showed only approximately 30% of the normal stomatal closure response, and was greatly slowed compared to the response in the presence of [Ca2+]i elevations. The ABA-insensitive mutants ost1-2, abi2-1 and gca2 showed partial stomatal closure responses that correlate with [Ca2+]i dependent ABA signaling. Interestingly, patch-clamp experiments showed that exposure of guard cells to ABA greatly enhances the ability of cytosolic Ca2+ to activate S-type anion channels and down-regulate inward-rectifying K+ channels, providing strong evidence for a Ca2+ sensitivity priming hypothesis. The present study demonstrates and quantifies an attenuated and slowed ABA response when [Ca2+]i elevations are directly inhibited in guard cells. A minimal model is discussed, in which ABA enhances (primes) the [Ca2+]i sensitivity of stomatal closure mechanisms. PMID- 19302419 TI - SnRK1 (SNF1-related kinase 1) has a central role in sugar and ABA signalling in Arabidopsis thaliana. AB - The proteins kinases SNF1/AMPK/SnRK1 are a subfamily of serine/threonine kinases that act as metabolite sensors to constantly adapt metabolism to the supply of, and demand for, energy. In the yeast Saccharomyces cerevisiae, the SNF1 complex is a central component of the regulatory response to glucose starvation. AMP activated protein kinase (AMPK) the mammalian homologue of SNF1, plays a central role in the regulation of energy homeostasis at the cellular as well as the whole body levels. In Arabidopsis thaliana, SnRK1.1 and SnRK1.2 have recently been described as central integrators of a transcription network for stress and energy signalling. In this study, biochemical analysis established SnRK1.1 as the major SnRK1 isoform both in isolated cells and leaves. In order to elucidate the function of SnRK1.1 in Arabidopsis thaliana, transgenic plants over-expressing SnRK1.1 were produced. Genetic, biochemical, physiological and molecular analyses of these plants revealed that SnRK1.1 is implicated in sugar and ABA signalling pathways. Modifications of the starch and soluble sugar content were observed in the 35S:SnRK1.1 transgenic lines. Our studies also revealed modifications of the activity of essential enzymes such as nitrate reductase or ADP-glucose pyrophosphorylase, and of the expression of several sugar-regulated genes, confirming the central role of the protein kinase SnRK1 in the regulation of metabolism. PMID- 19302420 TI - Lipid-induced ER stress: synergistic effects of sterols and saturated fatty acids. AB - Stress within the endoplasmic reticulum (ER) induces a coordinated response, namely the unfolded protein response (UPR), devoted to helping the ER cope with the accumulation of misfolded proteins. Failure of the UPR plays an important role in several human diseases. Recent studies report that intracellular accumulation of saturated fatty acids (SFAs) and cholesterol, seen in diseases of high incidence, such as obesity or atherosclerosis, results in ER stress. In the present study, we evaluated the effects of perturbations to lipid homeostasis on ER stress/UPR induction in the model eukaryote Saccharomyces cerevisiae. We show that SFA originating from either endogenous(preclusion of fatty acid desaturation) or exogenous (feeding with extracellular SFA) sources trigger ER stress and that ergosterol, the major sterol in yeast, acts synergistically with SFA in this process. This latter effect is connected to ergosterol accumulation within microsomal fractions from SFA-accumulating cells, which display highly saturated phospholipid content. Moreover, treating the cells with the molecular chaperone 4-phenyl butyrate abolishes UPR induction, suggesting that lipid induced ER stress leads to an overload of misfolded protein that acts, in turn, as the molecular signal for induction of the UPR. The present data are discussed in the context of human diseases that involve lipid deregulation. PMID- 19302421 TI - Labile hypertension: lessons to be learned from musical improvisation. PMID- 19302422 TI - Cost effectiveness analysis of a hypertension management program in patients with type 2 diabetes. AB - Hypertension is a costly disease; however, the investment needed for a cost neutral hypertension management program (HMP) is unknown. A Markov decision analytic model simulated the outcomes of a hypothetical HMP. Patients were between the ages of 25 and 65 years, had existing hypertension, and were newly diagnosed with diabetes. The control group received standard care. The HMP group received standard care and were enrolled in an HMP. Data regarding rates of disease states and costs were gathered from the literature. A third-party payer can invest as much as $159, $109, and $41 per person per month in an HMP for a neutral return on investment in the 5-year, 3-year, and 1-year time horizon, respectively. The HMP group achieved greater gains in quality-adjusted life-years and lower total health-related costs. As the time horizon increases, more money can be invested. HMPs can be a cost-effective and cost-neutral proposition. PMID- 19302423 TI - Therapeutic effect of an interval exercise training program in the management of erectile dysfunction in hypertensive patients. AB - The purpose of the present study was to determine the effect of an interval exercise training program on C-reactive protein and erectile dysfunction (ED) management in older male hypertensive patients. Twenty-two male hypertensive patients with ED (n=22; 62.10+/-5.23 years) were involved in interval exercise training (60%-79% heart rate max reserve) for 8 weeks for a duration of between 45 to 60 min/d, while an age-matched control hypertensive (n=21; 64.00+/-4.77 years) group remained sedentary during this period. The study revealed a significant effect of an exercise training program on erectile function of hypertensive patients with ED (P<.05). It was concluded that an exercise program is a possible effective noninvasive and nonpharmacologic management of ED in male hypertensive patients. PMID- 19302424 TI - Hypertension, insulin resistance, and aldosterone: sex-specific relationships. AB - African Americans, particularly men, have the highest morbidity and mortality rates from hypertension in the United States. The authors studied 527 African Americans in a general clinical research center to determine whether there are sex differences in the relationships between hypertension with insulin resistance (IR) and aldosterone, which are risk factors for cardiovascular disease. Measurements included ambulatory blood pressure (BP), anthropometric measures, plasma renin activity, plasma aldosterone (PA) concentration, and fasting serum lipids, glucose, and insulin. IR was estimated using the Homeostasis Model Assessment (HOMA) model. BP correlated with aldosterone in both sexes. However, both BP and PA correlated with IR in men, but not in women. Compared with men in the lower tertile of HOMA-IR, men in the upper tertile had higher mean systolic BP, a higher odds ratio of having hypertension, and higher levels of PA. The association of IR with both hypertension and PA in men, but not in women, may contribute to the high prevalence of cardiovascular disease in African American men. PMID- 19302425 TI - Awareness of hypertension and proteinuria in randomly selected patients in 11 Italian cities. A 2005 report of the National Kidney Foundation of Italy. AB - Arterial hypertension and proteinuria are risk factors for chronic kidney disease. A mobile clinic was parked in a central plaza of 11 Italian cities to check blood pressure (BP), prescribe antihypertensive drugs, assess for proteinuria, and provide awareness about hypertension. Among 3757 patients, 56% were hypertensive, 37% were not diabetic nor proteinuric with BP >or=140/90 mm Hg, 17% were diabetic or proteinuric with BP >or=130/80 mm Hg, and 11% were on treatment with BP at target. Among 1204 treated patients, 400 (33%) had controlled BP. Among all 2114 hypertensive patients, only 1344 (64%) were aware of their hypertension. Awareness was greater among treated patients at target (99%). As many as 523 (14%) patients had proteinuria >or=30 mg/dL. The authors conclude that awareness of people walking in the street about their BP and proteinuria is insufficient. Mobile screening clinics may increase public awareness and detection of hypertension and proteinuria in the general community and detect patients at risk for chronic kidney disease. PMID- 19302426 TI - The message for World Kidney Day 2009: hypertension and kidney disease: a marriage that should be prevented. AB - The prevalence of chronic kidney disease (CKD) continues to increase worldwide as does end stage renal disease. The most common, but not only, causes of CKD are hypertension and diabetes. CKD is associated with a significant increase in cardiovascular (CV) risk as most patients with CKD die of a CV cause. Moreover, CV risk increases proportionally as estimated glomerular filtration rate falls below 60 mL/min. CV causes of death in CKD are more prevalent than those from cancer; as a result, the identification and reduction of CKD is a public health priority. High blood pressure is a key pathogenic factor that contributes to the deterioration of kidney function. The presence of kidney disease is a common and underappreciated preexisting medical cause of resistant hypertension. Therefore, treatment of hypertension has become the most important intervention in the management of all forms of CKD. For this reason, World Kidney Day on March 12, 2009 will emphasize the role of hypertension. PMID- 19302427 TI - Hypertension in Asian/Pacific Island Americans. AB - Asian/Pacific Islander Americans (APIAs) are the fastest growing population in the United States by percentage. Hypertension is common and increases cardiovascular risk to a great extent in this population. The medical problems of this group are being increasingly encountered by US physicians. Many gene mutations associated with hypertension are more common in Asians. The significance of these polymorphisms in the pathogenesis of hypertension in APIAs is unclear. The percentage of APIAs who are aware, treated, and controlled is small. There may be some differences in the responses to antihypertensive medications between APIAs and whites. The results of human studies on the effect of drinking of tea on blood pressure in different groups are conflicting. Cough associated with angiotensin-converting enzyme inhibitor therapy may be more common in APIAs than in whites. There is a need for more education of APIAs regarding hypertension and for more effective treatment of hypertension by the physicians caring for this population. PMID- 19302428 TI - Commentary on the World Kidney Committee's position paper on hypertension and kidney disease. PMID- 19302429 TI - Assessing the blood pressure-lowering efficacy of antihypertensive medications: which blood pressures should we use? PMID- 19302430 TI - When the clinical trials raise more questions than answers: blood pressure controversy beyond the TRANSCEND results. PMID- 19302431 TI - Altitude-related hypertension. PMID- 19302434 TI - Hypertension associated with rhabdomyolysis may also be caused by unilateral adrenal hyperplasia. PMID- 19302435 TI - Effect of eicosapentaenoic acid on bone changes due to methylprednisolone in rats. AB - The present study was conducted to investigate the effects of eicosapentaenoic acid on glucocorticoid-induced bone changes in rats, and to compare them with those of alendronate. Thirty six male Wistar rats, 2.5 months of age, were divided into six groups (n = 6 each) and treated with 0.9% NaCl (control), methylprednisolone 7 mg/kg, once a week subcutaneously, methylprednisolone + alendronate 20 microg/kg, twice a week subcutaneously and methylprednisolone + 80 or 160 or 320 mg/kg eicosapentaenoic acid, per day orally, for 6 weeks. At the end of the experiment, serum and urinary parameters of bone metabolism determined and bone histomorphometric analyses performed on cancellous bone of femoral epiphysis and metaphysis and cortical bone of tibial diaphysis. There were no significant differences in serum and urinary parameters among groups. Decrease of epiphyseal and metaphyseal trabecular width, epiphyseal bone area/tissue area and increase of epiphyseal trabecular separation observed in the methylprednisolone group compared to control. Alendronate restored all of these parameters except metaphyseal trabecular width, which increased significantly by eicosapentaenoic acid at the doses of 80 and 160 mg/kg. Effects of alendronate and 160 mg/kg eicosapentaenoic acid on bone area/tissue area, alendronate and eicosapentaenoic acid at the doses of 80 and 160 mg/kg on trabecular separation and alendronate and eicosapentaenoic acid at doses of 160 and 320 mg/kg on epiphyseal trabecular width were statistically similar. Methylprednisolone did not significantly change cortical bone parameters including cortical width and marrow area/cortical area. Eicosapentaenoic acid, especially, at the dose of 160 mg/kg exerts beneficial effects on methylprednisolone-induced bone changes in rats; these effects are similar or sometimes even better than alendronate. PMID- 19302436 TI - Nausea and vomiting side effects with opioid analgesics during treatment of chronic pain: mechanisms, implications, and management options. AB - OBJECTIVES: Gastrointestinal (GI) side effects such as nausea and vomiting are common following opioid analgesia and represent a significant cause of patient discomfort and treatment dissatisfaction. This review examines the mechanisms that produce these side effects, their impact on treatment outcomes in chronic pain patients, and counteractive strategies. RESULTS: A number of mechanisms by which opioids produce nausea and vomiting have been identified. These involve both central and peripheral sites including the vomiting center, chemoreceptor trigger zones, cerebral cortex, and the vestibular apparatus of the brain, as well as the GI tract itself. Nausea and vomiting have a negative impact on treatment efficacy and successful patient management because they limit the effective analgesic dosage that can be achieved and are frequently reported as the reason for discontinuation of opioid pain medication or missed doses. While various strategies such as antiemetic agents or opioid switching can be employed to control these side effects, neither option is ideal because they are not always effective and incur additional costs and inconvenience. Opioid-sparing analgesic agents may provide a further alternative to avoid nausea and vomiting due to their reduced reliance on mu-opioid signalling pathways to induce analgesia. CONCLUSIONS: Nausea and vomiting side effects limit the analgesic efficiency of current opioid therapies. There is a clear need for the development of improved opioid-based analgesics that mitigate these intolerable effects. PMID- 19302437 TI - Reduction in medication costs for patients with chronic nonmalignant pain completing a pain rehabilitation program: a prospective analysis of admission, discharge, and 6-month follow-up medication costs. AB - OBJECTIVE: Chronic nonmalignant pain (CNMP) is both a prevalent and a costly health problem in our society. Pain rehabilitation programs have been shown to provide cost-effective treatment. A treatment goal for some rehabilitation programs is reduction in the use of pain-related medication. Medication costs savings from pain rehabilitation programs have not been analyzed in previous studies. DESIGN: This prospective cohort study of 186 patients with CNMP addresses the costs of medications at admission to a 3-week outpatient pain rehabilitation program, at discharge, and at 6-month follow-up. Medication use was determined through a detailed pharmacist interview with patients at admission and discharge. Patients were sent questionnaires 6 months after program completion, which obtained current medication information. RESULTS: Statistically significant medication cost savings were seen for program completers at discharge and at 6-month follow-up (P < 0.05). The mean (standard deviation) daily prescription medication cost reduction from admission to discharge was $9.31 ($12.70) using the average wholesale price of medications. From the original study cohort, 121 patients completed the 6-month follow-up survey. The mean daily prescription medication cost savings from admission to 6-month follow-up was $6.68 ($14.40). CONCLUSION: Patients benefited from significant medication cost savings at the completion of the 3-week outpatient pain rehabilitation program and maintained significant savings after 6 months. This study adds to the current literature on the economic value of comprehensive pain rehabilitation programs. PMID- 19302438 TI - Pulsed radiofrequency for chronic testicular pain-a preliminary report. AB - OBJECTIVE: To evaluate the effectiveness of pulsed radiofrequency (PRF) of spermatic cord in the treatment of chronic testicular pain. DESIGN: Ten patients with chronic testicular pain were treated with PRF stimulation of the spermatic cord. A radiofrequency probe placed percutaneously into the spermatic cord was used to deliver four 120-second cycles of 20-millisecond pulses at 2 Hz. Test stimulation was first used to confirm the precise placement of the probe. The short-form McGill Pain Questionnaire was used to assess pain before treatment and at 3 months. Patients who had experienced improvement were followed up by telephone, to determine whether pain relief was sustained. RESULTS: Ten patients were entered into the study but one was lost to follow-up. Of the nine patients evaluated, four had complete resolution of pain, while one had partial pain relief. Three patients experienced no change and one reported that his pain was worse. All patients who experienced complete and partial pain relief continued to do so at a mean long-term follow-up of 9.6 months (range 3-14 months). There were no complications observed immediately or during the follow-up period. CONCLUSION: In this pilot study, pain scores improved in five out of nine patients. PRF of spermatic cord appears to be a safe minimally invasive outpatient procedure that should be investigated further with placebo-controlled trials. PMID- 19302439 TI - Neurophysiological tools for small fiber assessment in painful diabetic neuropathy (comment letter). PMID- 19302441 TI - Stiff limb syndrome: end of spectrum or a separate entity? AB - BACKGROUND: Stiff-person syndrome is a rare disorder characterized by rigidity of axial or limb muscles with episodes of co-contraction of agonist and antagonist muscles during the spasms. In some patients axial or limb involvement may predominate and may have unusual manifestations. DESIGN: Case report. SETTING: Tertiary care teaching hospital. PATIENT: A 42-year-old farmer presented with seasonal occurrence of hiccup and vomiting during summer months for the last 3 years. He had painful lower limb spasms lasting for 2-3 minutes every 10-15 minutes for the past 20 days. His neurological examination was normal, erythrocyte sedimentation rate (ESR) was 50 mm at 1st hour, and cerebrospinal fluid protein 78 mg/dL without pleocytosis. Radiograph of chest, abdominal ultrasound, and craniospinal magnetic resonance imaging were normal. The patient improved on diazepam. CONCLUSION: Our patient is a forme fruste of stiff person syndrome with hiccups and vomiting due to diaphragmatic spasm. PMID- 19302442 TI - Hypoxia-inducible factor-dependent production of profibrotic mediators by hypoxic hepatocytes. AB - BACKGROUND/AIMS: During the development of liver fibrosis, mediators are produced that stimulate cells in the liver to differentiate into myofibroblasts and to produce collagen. Recent studies demonstrated that the transcription factor, hypoxia-inducible factor-1alpha (HIF-1alpha), is critical for upregulation of profibrotic mediators, such as platelet-derived growth factor-A (PDGF-A), PDGF-B and plasminogen activator inhibitor-1 (PAI-1) in the liver, during the development of fibrosis. What remains unknown is the cell type-specific regulation of these genes by HIF-1alpha in liver cell types. Accordingly, the hypothesis was tested that HIF-1alpha is activated in hypoxic hepatocytes and regulates the production of profibrotic mediators by these cells. METHODS: In this study, hepatocytes were isolated from the livers of control and HIF-1alpha- or HIF-1beta-deficient mice and exposed to hypoxia. RESULTS: Exposure of primary mouse hepatocytes to 1% oxygen stimulated nuclear accumulation of HIF-1alpha and upregulated PAI-1, vascular endothelial cell growth factor and the vasoactive peptides adrenomedullin-1 (ADM-1) and ADM-2. In contrast, the levels of PDGF-A and PDGF-B mRNAs were unaffected in these cells by hypoxia. Exposure of HIF 1alpha-deficient hepatocytes to 1% oxygen only partially prevented upregulation of these genes, suggesting that other hypoxia-regulated transcription factors, such as HIF-2alpha, may also regulate these genes. In support of this, HIF-2alpha was activated in hypoxic hepatocytes, and exposure of HIF-1beta-deficient hepatocytes to 1% oxygen completely prevented upregulation of PAI-1, vascular endothelial cell growth factor and ADM-1, suggesting that HIF-2alpha may also contribute to upregulation of these genes in hypoxic hepatocytes. CONCLUSIONS: Collectively, our results suggest that HIFs may be important regulators of profibrotic and vasoactive mediators by hypoxic hepatocytes. PMID- 19302443 TI - Antineoplastic effects of Magnoliae officinalis in tumours other than hepatomas. PMID- 19302444 TI - Neuropsychological assessment of hepatic encephalopathy: ISHEN practice guidelines. AB - Low-grade or minimal hepatic encephalopathy (MHE) is characterised by relatively mild neurocognitive impairments, and occurs in a substantial percentage of patients with liver disease. The presence of MHE is associated with a significant compromise of quality of life, is predictive of the onset of overt hepatic encephalopathy and is associated with a poorer prognosis for outcome. Early identification and treatment of MHE can improve quality of life and may prevent the onset of overt encephalopathy, but to date, there has been little agreement regarding the optimum method for detecting MHE. The International Society on Hepatic Encephalopathy and Nitrogen Metabolism convened a group of experts for the purpose of reviewing available data and making recommendations for a standardised approach for neuropsychological assessment of patients with liver disease who are at risk of MHE. Specific recommendations are presented, along with a proposed methodology for further refining these assessment procedures through prospective research. PMID- 19302445 TI - Time-dependent association between coagulation factor inactivation and increased elastase during experimental sepsis. PMID- 19302446 TI - Molecular mechanisms underlying hemophilia A phenotype in seven females. AB - BACKGROUND: Hemophilia A (HA) in females is a rare observation. Here we describe various genetic mechanisms that result in phenotypic expression of HA in seven females. METHODS: The F8 gene was examined in all patients and relatives by direct sequencing. Multiplex ligation-dependent probe amplification (MLPA) was performed for large deletion screening. X chromosome inactivation was studied by PCR analysis of a polymorphic CAG repeat in the first exon of the human androgen receptor (HUMARA) gene. RESULTS: In two females sequencing of the F8 gene revealed homozygous missense mutations (Arg593Cys and Tyr1680Phe) as a consequence of consanguineous marriage. The third case was due to compound heterozygosity comprising the missense mutation Leu412Phe inherited from the carrier mother, together with a de novo large deletion spanning exon 9-22, probably originating from the germ cells of the healthy father. Three further cases shared a common mechanism representing heterozygous mutations in the F8 gene (Arg1781His, Arg327His, small deletion in exon 10) combined with non-random inactivation of the X chromosome. The final case describes a coincidental inheritance of HA and Coffin-Lowry syndrome in the same family. The HA phenotype results from a heterozygous small deletion affecting the F8 gene (c.6872 del CT leading to Thr2272fs) and a complete inactivation of the maternal X chromosome, which segregates with Coffin-Lowry syndrome in the two brothers of the proposita. CONCLUSIONS: In conclusion, molecular genetic analysis represents an essentially valuable tool in elucidating the nature of the molecular mechanisms underlying the HA phenotype in females. PMID- 19302447 TI - Risk stratification and outcomes in hemodynamically stable patients with acute pulmonary embolism: a prospective, multicentre, cohort study with three months of follow-up. AB - BACKGROUND: The role of risk stratification in normotensive patients with acute pulmonary embolism (PE) is still unclear. OBJECTIVES: We evaluated, in these patients, the usefulness of six prognostic markers for predicting in-hospital adverse events related to PE and 3-month mortality. PATIENTS/METHODS: Two hundred and one consecutive patients with confirmed acute PE and normal blood pressure, who were administered conventional anticoagulation, were recruited in a multicentre prospective cohort study with 3 months of follow-up. At baseline, they received a comprehensive risk-evaluation including echocardiographic assessment of right ventricular dysfunction, determination of troponin I, brain natriuretic peptide and D-dimer, arterial blood gas analysis and a clinical score. Primary outcome of the study was PE-related in-hospital death or clinical deterioration. Secondary outcomes were in-hospital and 3-month all-cause mortality. RESULTS: The primary outcome occurred in one patient (0.5%), who died from PE during hospitalization. The in-hospital and 3-month all-cause mortality were 2% and 9%, respectively. None of the prognostic markers was predictive of the primary outcome. Clinical score, troponin I and hypoxemia predicted in hospital all-cause mortality (P = 0.02, 0.01 and < 0.01, respectively). Clinical score (HR, 4.7; 95% CI, 1.9-12.0), D-dimer (4.8; 1.4-16.3), hypoxemia (5.7; 2.1 15.1) and troponin I (7.5; 2.5-22.7) were predictors of 3-month all-cause mortality on univariate analysis. On multivariate analysis clinical score and troponin I remained independently predictive. CONCLUSIONS: We did not find prognostic markers useful as predictors of in-hospital PE-related adverse events. Clinical score, troponin I and hypoxemia predicted in-hospital all-cause mortality. Clinical score and troponin I independently predicted 3-month all cause mortality. PMID- 19302448 TI - 'How did you validate ...?' A plea for validation studies. PMID- 19302449 TI - Towards better blood transfusion--recruitment and training. AB - Recent national initiatives in blood transfusion safety in the UK have created the need for an expansion in haematologists subspecializing (wholly or in part) in transfusion medicine. In 2008, there are 62 transfusion consultants in the UK, but only 42 are full time, and only 19 have hospital sessions. Despite the need for expansion, recruitment appears difficult. The English blood transfusion service, National Health Service Blood and Transplant (NHSBT), is undergoing major reconfiguration, and the current practice of transfusion training for haematology specialists primarily at blood centres with little or no hospital training is not sustainable or desirable. Delivering a high-quality transfusion programme to haematology trainees is best achieved through an increased emphasis on hospital-based training. Improved research opportunities, joint NHSBT/hospital posts and a separate subspecialty training curriculum may stimulate interest in transfusion medicine. PMID- 19302450 TI - Drugs and blood transfusions: dogma- or evidence-based practice? AB - There is a lack of consensus on the safety of the coadministration of drugs and red blood cells (RBCs). A systematic review was undertaken to establish the evidence base for this question and assess how the evidence may be translated into present clinical day practice. Comprehensive searches of MEDLINE, EMBASE, CINAHL, the Cochrane Library and hand searching of transfusion journals, guidelines and websites identified 12 relevant papers: 11 in-vitro experiments and 1 case report. Data on incidences of haemolysis and agglutination following coadministration were extracted and analysed. Overall findings suggest that iron chelators (two papers), antimicrobials (three papers) and lower doses of opioids (three papers) are safe to coadminister with RBCs. Haemolysis was observed with higher doses of opioids (three papers). Transposition of these findings to clinical practice is limited because of the lack of clinical applicability of in vitro experiments and diversity in how, and what, clinical outcome measures were used. Further evidence from true clinical settings would be required to inform clinical practice on the efficacy and safety of the coadministration of drugs and RBCs. PMID- 19302451 TI - Chagas' disease: lessons from routine donation testing. AB - American Trypanosomiasis, or Chagas' disease, is a parasitic infection caused by the protozoan Trypanosoma cruzi. It is endemic in a large area of the American continent, extending from Mexico to Argentina. Imported Chagas' disease is now appearing as a new threat to non-endemic countries, mostly because of a steady increase of foreign residents from Latin America. Chagas' disease becomes chronic in the vast majority of infected individuals. This finding gives rise to problems for blood transfusion services. In non-endemic countries, transfusion is the most likely infection route. Strategies to reduce transmission by transfusion include blood donor selection and deferral, blood donation testing, leukoreduction, filters and pathogen inactivation systems. Policies in endemic and non-endemic areas are quite different: in endemic countries, universal screening of blood donations for T. cruzi antibody detection is mandatory. However, in non-endemic countries, there are two different approaches: one is the deferral of people at risk of Chagas' disease and the second approach is to accept the blood donation if specific laboratory assay results are negative. This second approach is being introduced in countries where there is a substantial Latin American population, such as United States, Spain and France. The assays used for the detection of T. cruzi are mostly immunologic, made with T. cruzi antigen homogenates or with recombinant antigens. Complementary assays, such as indirect immunofluorescence (IIF) or immunoblot, can help ascertain antibody specificity. PMID- 19302452 TI - The introduction of anti-HTLV testing of blood donations and the risk of transfusion-transmitted HTLV, UK: 2002-2006. AB - The objectives of the study were to describe the introduction of testing blood donations for antibodies to human T-cell lymphotropic virus (anti-HTLV) and to determine the risk of HTLV potentially infectious donations entering the UK blood supply. The rationale for testing was based on (i) evidence of transmission through transfusion in the UK, (ii) the serious nature of HTLV I-associated morbidity and (iii) evidence of infection in UK blood donors. From mid-2002, all blood donations made at UK blood centres were tested in pooled samples using Abbott-Murex HTLV I/II GE 80/81 enzyme immunoassay (EIA). Surveillance data were used to calculate the incidence and prevalence of anti-HTLV and derive estimates of risk. Between August 2002 and December 2006, 106 donations were confirmed positive for anti-HTLV (95 anti-HTLV I and 11 anti-HTLV II). Prevalence was 10 fold higher among donations from new donors than repeat (4.0 and 0.42 per 100 000 donations), and only one repeat donor had evidence of seroconversion. The risk of an HTLV I potentially infectious donation entering the UK blood supply was estimated at 0.11 per million donations (95% confidence interval 0.06 to 0.18). The current very low observed incidence and prevalence among blood donors reflect the very low estimated risk of an HTLV I-positive donation entering the UK blood supply. A change in either the epidemiology of HTLV in UK blood donors or the length of the window period of the test should prompt further review of the risk and a reassessment of anti-HTLV testing in the UK. PMID- 19302453 TI - An individualized pre-operative blood saving protocol can increase pre-operative haemoglobin levels and reduce the need for transfusion in elective total hip or knee arthroplasty. AB - We have prospectively evaluated the efficacy of an individualized pre-operative blood saving protocol in elective total hip arthroplasty (THA) or total knee arthroplasty (TKA). The primary aim was to obtain a pre-operative haemoglobin (Hb) level of > or =14 g dL(-1). A reduction in requirements for allogeneic transfusion was considered the second aim. Several strategies are available for increasing pre-operative Hb levels and reducing red blood cell (RBC) transfusions following THA or TKA, but the success of these programmes depends on selecting the most appropriate treatment for each patient. Three hundred and five patients with an indication of elective THA or TKA were individually assigned to the following strategies according to Hb and ferritin levels and medical conditions: (a) no pre-operative intervention, (b) oral iron therapy, (c) intravenous (i.v.) iron therapy, (d) recombinant human erythropoietin alpha with i.v. iron and (e) pre-operative autologous donation (PAD) plus oral iron. Eighty-two percent of the patients reached a pre-operative Hb level of > or =14 g dL(-1) compared with 62% of patients with Hb levels of > or =14 g dL(-1) at the baseline visit. Treatment with PAD showed a significant reduction in the pre-operative Hb levels. The rate of RBC transfusion was 18.8% compared with 31.5% of matched historic group (P < 0.001). In conclusion, all patients scheduled to undergo THA or TKA should be candidates for an individualized pre-operative blood salvage programme. PMID- 19302454 TI - Recombinant activated factor VII in the treatment of non-haemophilia patients: physician under-reporting of thromboembolic adverse events. AB - The objective of this study was to determine if clinically important thromboembolic adverse events (TAEs) because of recombinant activated factor VII (rFVIIa) administration are being under-reported. rFVIIa is a potent haemostatic agent with a short half-life of 2.6 h that is increasingly used in 'off-label' situations. Retrospective review of 94 patients who received rFVIIa during 1 January 2003 to 30 June 2007 was carried out at a tertiary care centre. Sixty nine patients, 32 females and 37 males, mean age 55 years (18-84 years), satisfied study criteria of off-label usage. This was a high-risk population with 33 (48%) deaths. A mean dose of 8.2 mg (2.4-19.2 mg) was administered in two average divided doses. Thirty-six potential TAEs were identified in 29 patients, and of these, 12 patients had TAEs deemed to be rFVIIa related and were identified on average 8.8 days after exposure to rFVIIa. Forty-eight (70%) physician questionnaires were completed; however, no TAEs were reported in these questionnaires or on chart review. Potential clinically significant TAEs are being under-reported by treating physicians. Until further evidence, we suggest the urgent need to develop consensus recommendations for utilization and required follow up to monitor the safety of rFVIIa and that at a minimum, all use of rFVIIa should be regulated through a gate-keeping mechanism that ensures adherence to these policies. Furthermore, prospective registries and trials are necessary to evaluate the efficacy and safety of rFVIIa in off-label settings. PMID- 19302455 TI - Management of emergency cardiac surgery in a patient with alloanti-Ge2. AB - Transfusion management of patients alloimmunized against high-prevalence erythrocyte antigens is often problematic in emergency situations. In these patients, incompatible transfusion may be less harmful for the patient than delaying surgery, especially if the antibody is not clinically significant. We report an untransfused 75-year-old Caucasian man (blood group O) with an alloantibody against the Gerbich-2 (Ge2) antigen who required emergency cardiac surgery. Because cross-match compatible blood was not available in an acceptable timeframe, we performed a 'biological cross-match' with sequential transfusion of 20 and 50 mL and then the entire unit of incompatible red blood cells (RBCs) before surgery. When there were no clinical symptoms of adverse biological effects, we transfused two further incompatible packed RBCs during surgery. Subsequently, there was neither clinical nor laboratory evidence of major intra- or extravascular haemolysis, suggesting that this anti-Ge2 antibody was not clinically significant. PMID- 19302456 TI - HPA-1 allelic frequencies in the Greek population. PMID- 19302457 TI - Distribution of the human platelet alloantigen HPA-17w allele in the Chinese Han population. PMID- 19302458 TI - Combined antifungal therapy, iron chelation and surgical resection as treatment of hepatic zygomycosis in a patient with haematological malignancy. AB - We describe the case of a 19-year-old boy with acute leukaemia who developed primary hepatic zygomycosis. The patient presented with febrile neutropenia and severe abdominal tenderness. Despite the administration of antibiotics and liposomal Amphotericin-B (L-AmB), the CT scan demonstrated an increase in the size of liver lesions. A wide surgical resection was carried out and liver specimens demonstrated a branching, filamentous fungus that was identified as Rhizomucor pusillus by both phenotypic and molecular methods. The patient was treated with L-AmB combined with posaconazole, and deferasirox was subsequently added given the potential synergistic effect of this iron chelator in combination with L-AmB. Three months after surgical intervention, an allogeneic stem-cell transplantation was successfully carried out. The present case confirms that an early surgical management combined with antifungal agents is crucial to optimise the outcome of patients with zygomycosis and the use of deferasirox is a promising alternative. PMID- 19302459 TI - Antifungal activity of Pterocaulon alopecuroides (Asteraceae) against chromoblastomycosis agents. AB - Plants of the genus Pterocaulon (Asteraceae) are popularly used in the treatment of skin diseases caused by fungi and bacteria. The aim of this work was to investigate the in vitro activity of the crude methanolic extract obtained from the aerial parts of Pterocaulon alopecuroides (Lam.) against some agents of chromoblastomycosis, a chronic fungal infection of the skin and of the subcutaneous tissue caused by traumatic inoculation of the aetiological agent. The extract was active against all the strains tested showing a minimum inhibitory concentration between 625 and 2500 microg ml(-1). The assessment of fungistatic/fungicidal activity demonstrated that the extract was fungistatic against Fonsecaea spp. and fungicidal against all the other fungi. Our results indicate that the identification of bioactive components present in the crude methanolic extract of P. alopecuroides against chromoblastomycosis agents can be an important strategy to manage this mycosis in the future. PMID- 19302460 TI - Identification of an invasive infection of R. oryzae in a haematological patient using a molecular technique. AB - We report a case of cerebral mucormycosis in a 28-year-old male who was affected by chronic myeloid leukaemia and underwent allogeneic bone marrow transplantation. Nine months post-transplantation, he was admitted to the hospital with fever, bilateral eyelid oedema and neutropenia. X-ray analysis showed numerous areas of pulmonary parenchymal thickening, and a computed tomography scan of the brain showed inflammation of the frontal, maxillary, ethmoidal and sphenoidal sinuses and diffuse swelling of the periorbital tissues. Sinus cultures were taken, and based on its characteristic rhizoid structure, we classified the isolated fungus as a member of the genus Rhizopus. The fungus was identified as an Rhizopus oryzae species, as assessed by sequencing of the internal transcribed spacer of the rRNA gene. Treatment with amphotericin B was ineffective, however, and the patient died 2 weeks after admission. This case highlights the potential severity of an invasive infection of R. oryzae, identified by molecular biology techniques. PMID- 19302461 TI - Tinea capitis among children in the Columbus area, Ohio, USA. AB - Tinea capitis is a fungal infection of the hair follicles of the scalp. In the US, the most common organisms have traditionally been Trichophyton tonsurans, and occasionally Microsporum canis. This study was designed to examine patterns of organisms causing tinea capitis and determine factors associated with infection. A retrospective database analysis was conducted to locate records of patients with tinea capitis from May 2001 to May 2006 at Nationwide Children's Hospital in Columbus, OH. Descriptive statistics, frequency analysis, chi-squared test, and Student's t-test were performed to evaluate types of causative organisms and associated patient characteristics. One hundred and eighty-nine charts of patients with a positive scalp culture for tinea capitis were located. Trichophyton tonsurans (88.9%) was the foremost causative agent followed by Trichophyton violaceum (4.2%). Tinea capitis was more prevalent among African Americans and was more common in urban areas (P < 0.05). Children of African descent inhabiting urban settings were most vulnerable to tinea capitis. The most common organism isolated in this retrospective study was T. tonsurans. Trichophyton violaceum and Trichophyton soudanense were also isolated, which are not commonly reported causes of tinea capitis in the US. PMID- 19302462 TI - In vitro antifungal activity of 2-(2'-hydroxy-5'-aminophenyl)benzoxazole in Candida spp. strains. AB - The development of azole antifungals has allowed for the treatment of several fungal infections. However, the use of these compounds is restricted because of their hepatotoxicity or because they need to be administered together with other drugs in order to prevent resistance to monotherapy. Benzoxazole derivatives are among the most thriving molecular prototypes for the development of antifungal agents. 2-(2'-hydroxyphenyl) benzoxazoles are versatile molecules that emit fluorescence and have antibacterial, antiviral and antifungal properties. 2-(2' hydroxy-5'-aminophenyl) benzoxazole (HAMBO) was tested against Candida yeast. The inhibition provided by HAMBO was lower than that of fluconazole, showing low antifungal activity against Candida spp., but equivalent to that of benzoxazoles tested in similar studies. HAMBO showed fungistatic activity against all analysed strains. This class of novel benzoxazole compounds may be used as template to produce better antifungal drugs. PMID- 19302463 TI - Flexible social organization and high incidence of drifting in the sweat bee, Halictus scabiosae. AB - The very diverse social systems of sweat bees make them interesting models to study social evolution. Here we focus on the dispersal behaviour and social organization of Halictus scabiosae, a common yet poorly known species of Europe. By combining field observations and genetic data, we show that females have multiple reproductive strategies, which generates a large diversity in the social structure of nests. A detailed microsatellite analysis of 60 nests revealed that 55% of the nests contained the offspring of a single female, whereas the rest had more complex social structures, with three clear cases of multiple females reproducing in the same nest and frequent occurrence of unrelated individuals. Drifting among nests was surprisingly common, as 16% of the 122 nests in the overall sample and 44% of the nests with complex social structure contained females that had genotypes consistent with being full-sisters of females sampled in other nests of the population. Drifters originated from nests with an above average productivity and were unrelated to their nestmates, suggesting that drifting might be a strategy to avoid competition among related females. The sex specific comparison of genetic differentiation indicated that dispersal was male biased, which would reinforce local resource competition among females. The pattern of genetic differentiation among populations was consistent with a dynamic process of patch colonization and extinction, as expected from the unstable, anthropogenic habitat of this species. Overall, our data show that H. scabiosae varies greatly in dispersal behaviour and social organization. The surprisingly high frequency of drifters echoes recent findings in wasps and bees, calling for further investigation of the adaptive basis of drifting in the social insects. PMID- 19302464 TI - Species-wide phylogeography of North American mule deer (Odocoileus hemionus): cryptic glacial refugia and postglacial recolonization. AB - Quaternary climatic oscillations greatly influenced the present-day population genetic structure of animals and plants. For species with high dispersal and reproductive potential, phylogeographic patterns resulting from historical processes can be cryptic, overshadowed by contemporary processes. Here we report a study of the phylogeography of Odocoileus hemionus, a large, vagile ungulate common throughout western North America. We examined sequence variation of mitochondrial DNA (control region and cytochrome b) within and among 70 natural populations across the entire range of the species. Among the 1766 individual animals surveyed, we recovered 496 haplotypes. Although fine-scale phylogenetic structure was weakly resolved using phylogenetic methods, network analysis clearly revealed the presence of 12 distinct haplogroups. The spatial distribution of haplogroups showed a strong genetic discontinuity between the two morphological types of O. hemionus, mule deer and black-tailed deer, east and west of the Cascade Mountains in the Pacific Northwest. Within the mule deer lineage, we identified several haplogroups that expanded before or during the Last Glacial Maximum, suggesting that mule deer persisted in multiple refugia south of the ice sheets. Patterns of genetic diversity within the black-tailed deer lineage suggest a single refugium along the Pacific Northwest coast, and refute the hypothesis that black-tailed deer persisted in one or more northern refugia. Our data suggest that black-tailed deer recolonized areas in accordance with the pattern of glacial retreat, with initial recolonization northward along a coastal route and secondary recolonization inland. PMID- 19302465 TI - Do landscape processes predict phylogeographic patterns in the wood frog? AB - Understanding factors that influence population connectivity and the spatial distribution of genetic variation is a major goal in molecular ecology. Improvements in the availability of high-resolution geographic data have made it increasingly possible to quantify the effects of landscape features on dispersal and genetic structure. However, most studies examining such landscape effects have been conducted at very fine (e.g. landscape genetics) or broad (e.g. phylogeography) spatial scales. Thus, the extent to which processes operating at fine spatial scales are linked to patterns at larger scales remains unclear. Here, we test whether factors impacting wood frog dispersal at fine spatial scales are correlated with genetic structure at regional scales. Using recently developed methods borrowed from electrical circuit theory, we generated landscape resistance matrices among wood frog populations in eastern North America based on slope, a wetness index, land cover and absolute barriers to wood frog dispersal. We then determined whether these matrices are correlated with genetic structure based on six microsatellite markers and whether such correlations outperform a landscape-free model of isolation by resistance. We observed significant genetic structure at regional spatial scales. However, topography and landscape variables associated with the intervening habitat between sites provide little explanation for patterns of genetic structure. Instead, absolute dispersal barriers appear to be the best predictor of regional genetic structure in this species. Our results suggest that landscape variables that influence dispersal, microhabitat selection and population structure at fine spatial scales do not necessarily explain patterns of genetic structure at broader scales. PMID- 19302466 TI - Perspectives and challenges in landscape genetics. AB - A recent workshop held at the University of Grenoble gathered the leading experts in the field of landscape genetics and spatial statistics. Landscape genetics was only recently defined as an independent research field. It aims to understand the processes of gene flow and local adaptation by studying the interactions between genetic and spatial or environmental variation. This workshop discussed the perspectives and challenges of combining emerging molecular, spatial and statistical tools to unravel how landscape and environmental variables affect genetic variation. PMID- 19302467 TI - Not just vicariance: phylogeography of a Sonoran Desert euphorb indicates a major role of range expansion along the Baja peninsula. AB - To examine the generality of population-level impacts of ancient vicariance identified for numerous arid-adapted animal taxa along the Baja peninsula, we tested phylogeographical hypotheses in a similarly distributed desert plant, Euphorbia lomelii (Euphorbiaceae). In light of fossil data indicating marked changes in the distributions of Baja floristic assemblages throughout the Holocene and earlier, we also examined evidence for range expansion over more recent temporal scales. Two classes of complementary analytical approaches - hypothesis-testing and hypothesis-generating - were used to exploit phylogeographical signal from chloroplast DNA sequence data and genotypic data from six codominant nuclear intron markers. Sequence data are consistent with a scenario of mid-peninsular vicariance originating c. 1 million years ago (Ma). Alternative vicariance scenarios representing earlier splitting events inferred for some animals (e.g. Isthmus of La Paz inundation, c. 3 Ma; Sea of Cortez formation, c. 5 Ma) were rejected. Nested clade phylogeographical analysis corroborated coalescent simulation-based inferences. Nuclear markers broadened the temporal spectrum over which phylogeographical scenarios could be addressed, and provided strong evidence for recent range expansions along the north-south axis of the Baja peninsula. In contrast to previous plant studies in this region, however, the expansions do not appear to have been in a strictly northward direction. These findings contribute to a growing appreciation of the complexity of organismal responses to past climatic and geological changes - even when taxa have evolved in the same landscape context. PMID- 19302468 TI - Diversity despite dispersal: colonization history and phylogeography of Hawaiian crab spiders inferred from multilocus genetic data. AB - The Hawaiian archipelago is often cited as the premier setting to study biological diversification, yet the evolution and phylogeography of much of its biota remain poorly understood. We investigated crab spiders (Thomisidae, Mecaphesa) that demonstrate contradictory tendencies: (i) dramatic ecological diversity within the Hawaiian Islands, and (ii) accompanying widespread distribution of many species across the archipelago. We used mitochondrial and nuclear genetic data sampled across six islands to generate phylogenetic hypotheses for Mecaphesa species and populations, and included penalized likelihood molecular clock analyses to estimate arrival times on the different islands. We found that 17 of 18 Hawaiian Mecaphesa species were monophyletic and most closely related to thomisids from the Marquesas and Society Islands. Our results indicate that the Hawaiian species evolved from either one or two colonization events to the archipelago. Estimated divergence dates suggested that thomisids may have colonized the Hawaiian Islands as early as ~10 million years ago, but biogeographic analyses implied that the initial diversification of this group was restricted to the younger island of Oahu, followed by back colonizations to older islands. Within the Hawaiian radiation, our data revealed several well-supported genetically distinct terminal clades corresponding to species previously delimited by morphological taxonomy. Many of these species are codistributed across multiple Hawaiian Islands and some exhibit genetic structure consistent with stepwise colonization of islands following their formation. These results indicate that dispersal has been sufficiently limited to allow extensive ecological diversification, yet frequent enough that interisland migration is more common than speciation. PMID- 19302469 TI - Social support and intellectual disabilities: a comparison between social networks of adults with intellectual disability and those with physical disability. AB - BACKGROUND: Social support has been identified as a major protective factor in preventing mental health problems and also as a major contributor to quality of life. People with intellectual disabilities (ID) have been identified as having limited social support structures. Interventions have been focused on promoting their social presence and integration. However, previous studies have shown that this does not always lead to the formation of social relationships. To date few studies have looked at how having an ID leads to impoverished social networks. This study aimed to do this by contrasting the social relationships of people with physical disabilities (PD) and people with ID. METHODS: Two groups of participants were recruited; 30 people with mild ID and 17 people with PD. Social and functional support networks were assessed, in addition to life experiences. Between and within group differences were then explored statistically. RESULTS: Adults with ID had more restricted social networks than PD, despite being involved in more activities. Social support for adults with ID was mainly provided by family and carers and few relationships with non-disabled people were identified. In contrast adults with PD had larger social networks than had been reported in the mainstream literature and had a balance of relationships with disabled and non-disabled people. CONCLUSIONS: The results suggest that there are additional processes attached to having an ID, which lead to continued impoverished lifestyles. The findings also endorse other work that suggests being physically integrated and engaged in a wide range of activities does not guarantee good social and emotional support. PMID- 19302470 TI - Water intake and risk of hyponatraemia in Prader-Willi syndrome. AB - BACKGROUND AND METHODS: Unusual water intake and drinking behaviour has occasionally been observed in individuals with Prader-Willi syndrome (PWS). The aim of this study is to explore whether this observation is a part of the PWS phenotype and what the consequences may be. The parents of 51 individuals with PWS (age range 2-40 years) were asked by questionnaire to answer on past and present water intake, drinking behaviour, fluid preferences and medical treatment in their PWS-affected and unaffected children. Questionnaires with information on 47 PWS individuals and 17 without PWS were returned for analysis. The questionnaire information was complemented with information from the individual's medical records. Siblings to PWS individuals made up the control group. The study was approved by the regional medical research ethics committee. RESULTS: During infancy, 36 (76%) individuals with PWS disliked water without any flavouring and had an extremely small daily intake of water. Seven individuals (15%) increased the daily water intake to unusually high amounts. In 45 the clinical PWS diagnosis was confirmed by molecular (genetic) testing: nine of them with a confirmed PWS diagnosis had a deletion of chromosome 15q11-13, in nine individuals no deletion was identified. The majority of individuals who increased their water consumption to extreme values belonged to the non-deletion group. Two in the non-deletion group developed hyponatraemia while receiving psychiatric medication. CONCLUSIONS: Infants with PWS seem to be predisposed to unusual drinking behaviour. They dislike and have an unusually small intake of pure water without flavouring, and most of them continue this even after infancy. Some individuals, especially those without deletion, increase their fluid intake and also accept pure water. They have an increased risk of developing water retention and severe hyponatraemia if exposed to medication known to cause side effects like the syndrome of inappropriate antidiuretic hormone secretion. Perhaps this behaviour is just secondary to overeating; perhaps it is a result of a dysfunction of the hypothalamic nuclei engaged in antidiuretic hormone production. PMID- 19302471 TI - How employment support and social integration programmes are viewed by the intellectually disabled. AB - BACKGROUND: Supported employment is intended to facilitate and promote participation and integration of intellectually disabled citizens in society. This study investigated their view of the programme. METHODS: Q-methodology was used. Eighteen respondents with a mild intellectual disability rank-ordered 22 statements representing five main aspects of supported employment. The data were factor-analysed to group respondents according to their views. Q-methodology was a feasible approach that facilitated in-depth conversations with respondents with a mild intellectual disability in a playful manner. RESULTS: Two views on the impact of supported employment on social integration were observed: 'work as participation' and 'work as structure'. The first placed greater value on participation, task variety, belonging, and feeling appreciated; the second placed greater value on working independently, clear working agreements, and friendly co-workers. The views indicate two distinct approaches to effecting a positive relationship between supported employment and social integration. CONCLUSION: From the perspective of people with an intellectual disability supported employment contributes to self-development and has a positive effect on well-being, albeit in different ways for the individual groups. PMID- 19302472 TI - A longitudinal study of motivation and competence in children with Down syndrome: early childhood to early adolescence. AB - BACKGROUND: Motivation has been identified as an area of difficulty for children with Down syndrome. Although individual differences in mastery motivation are presumed to have implications for subsequent competence, few longitudinal studies have addressed the stability of motivation and the predictive validity of early measures for later academic achievement, especially in atypical populations. METHOD: The participants were 25 children with Down syndrome. Mastery motivation, operationalized as persistence, was measured in early childhood and adolescence using tasks and parent report. At the older age, preference for challenge, another aspect of mastery motivation, was also measured and the children completed assessments of academic competence. RESULTS: There were significant concurrent correlations among measures of persistence at both ages, and early task persistence was associated with later persistence. Persistence in early childhood was related to academic competence in adolescence, even when the effects of cognitive ability at the younger age were controlled. CONCLUSIONS: For children with Down syndrome, persistence appears to be an individual characteristic that is relatively stable from early childhood to early adolescence. The finding that early mastery motivation is significant for later achievement has important implications for the focus of early interventions. PMID- 19302473 TI - Family quality of life among families with a member who has an intellectual disability: an exploratory examination of key domains and dimensions of the revised FQOL Survey. AB - BACKGROUND: The Family Quality of Life Survey (FQOLS-2006) was developed as the result of increased interest in family quality of life (FQOL) among families with a member who has an intellectual disability (ID). The instrument includes nine life domains and six dimensions reflecting the main areas and characteristics of FQOL. The aim of the current study was to provide a descriptive analysis of the domains and dimensions of the survey and to explore their relationship to one another and to global satisfaction. METHOD: A convenience sample of 35 participants with a family member who had ID completed the FQOLS-2006 in a large urban centre in Canada. The data were analysed using descriptive analyses. RESULTS: The findings showed that although participants differentiated between different domains and dimensions, as reflected in their variability, stability was also found. For example, support from others was rated lowest across most dimensions, while family relationships and health of the family were generally rated higher. CONCLUSIONS: The findings strengthen the importance of examining both the overall perception of FQOL as well as the perceptions in individual life domains. Moreover, they highlight the need to examine each life domain according to its various dimensions. In sum, the results call for further examination of the FQOLS-2006 as a useful tool for assessing FQOL and for implementing services based on it. PMID- 19302474 TI - The atypical protein kinase C in Aplysia can form a protein kinase M by cleavage. AB - In vertebrates, a brain-specific transcript from the atypical protein kinase C (PKC) zeta gene encodes protein kinase M (PKM) zeta, a constitutively active kinase implicated in the maintenance of synaptic plasticity and memory. We have cloned the atypical PKC from Aplysia, PKC Apl III. We did not find a transcript in Aplysia encoding PKMzeta, and evolutionary analysis of atypical PKCs suggests formation of this transcript is restricted to vertebrates. Instead, over expression of PKC Apl III in Aplysia sensory neurons leads to production of a PKM fragment of PKC Apl III. This cleavage was induced by calcium and blocked by calpain inhibitors. Moreover, nervous system enriched spliced forms of PKC Apl III show enhanced cleavage. PKC Apl III could also be activated through phosphorylation downstream of phosphoinositide 3-kinase. We suggest that PKM forms of atypical PKCs play a conserved role in memory formation, but the mechanism of formation of these kinases has changed over evolution. PMID- 19302475 TI - The novel NMDA receptor antagonist, 2-hydroxy-5-(2,3,5,6-tetrafluoro-4 trifluoromethyl-benzylamino)-benzoic acid, is a gating modifier in cultured mouse cortical neurons. AB - Neu2000 [NEU, 2-hydroxy-5-(2,3,5,6-tetrafluoro-4-trifluoromethyl-benzylamino) benzoic acid], a derivative of sulfasalazine, attenuates NMDA-induced neuronal toxicity. Here we investigated the effects of NEU on the NMDA receptor (NMDAR) using whole-cell patch clamp technique to determine the molecular mechanisms underlying its neuroprotective role. NEU reversibly suppressed NMDA responses in an uncompetitive manner with fast binding kinetics. Its inhibition of NMDAR activity depended on both the concentration and the use of agonist but not on the membrane potential. NEU accelerated NMDA desensitization without affecting the binding affinity of NMDAR for its agonists and stabilized the closed state of NMDAR. Therefore, NEU should effectively alleviate disorders that are a result of glutamate excitoxicity with fewer side effects because it is a low-affinity gating modifier that antagonizes NMDAR in an uncompetitive manner. Moreover, in the presence of ifenprodil (an NR2B antagonist) but not NVP-AAM077 [(R)-[(S)-1-(4 bromo-phenyl)-ethylamino]-(2,3-dioxo-1,2,3,4-tetrahydro-quinoxalin-5-yl)-methyl] phosphonic acid, an NR2A antagonist], the extent of NEU block was decreased, suggesting that NEU is an NR2B-specific antagonist. PMID- 19302476 TI - CXCL12 increases human neural progenitor cell proliferation through Akt-1/FOXO3a signaling pathway. AB - CXCL12, a ligand for the chemokine receptor CXCR4, is well known in mediating neural progenitor cell (NPC) migration during neural development. However, the effects of CXCL12 on human NPC proliferation and its associated signaling pathways remain unclear. The transcription factor, FOXO3a, a downstream target of Akt-1, is critical for cell cycle control and may also play an important role in regulating NPC proliferation. In this study, we found that CXCL12 promotes human NPC proliferation as determined by the proliferation marker Ki67 and BrdU incorporation. This CXCL12-mediated NPC proliferation was associated with an increase in Akt-1 and FOXO3a phosphorylation in a time- and dose-dependent manner. The CXCR4 antagonist (T140) or inhibitors for G proteins (Pertussis toxin) and phosphoinositide 3-kinase (PI3K) (LY294002) abolished CXCL12-mediated NPC proliferation and phosphorylation of Akt-1 and FOXO3a. The roles of Akt-1 and FOXO3a in CXCL12-mediated NPC proliferation were further investigated by using adenoviral over-expression in NPCs. Over-expression of dominant-negative Akt-1 or wild-type FOXO3a in NPC abrogated CXCL12-mediated proliferation. These data suggest that CXCL12-mediated NPC proliferation is reliant upon the phosphorylation of Akt-1 and FOXO3a and gives insight to an essential role of CXCL12 in neurogenesis. Understanding this mechanism may facilitate the development of novel therapeutic targets for NPC proliferation during neurogenesis. PMID- 19302477 TI - Use of a custom-designed adapter to manage ICD lead fracture. AB - We report the case of a 72-year-old patient with ischemic cardiomyopathy who was found to have a lead fracture in the RV coil DF-1 connector of an integrated bipolar implantable cardioverter-defibrillator (ICD) lead system at the time of upgrade to a biventricular ICD. Due to the redundant connection of the RV coil to the DF-1 connector and the ring on the IS-1 connector, a custom-designed adapter was used to exclude the site of fracture. This novel solution spared the patient the potential risks of lead extraction or abandoning the fractured ICD lead. In this way, pacing and defibrillating capabilities were fully restored, providing the patient 6 years of optimal device functioning. PMID- 19302478 TI - Using the 12-lead ECG to localize the origin of atrial and ventricular tachycardias: part 2--ventricular tachycardia. AB - Monomorphic ventricular tachycardia (VT) can arise from multiple different ventricular locations in the context of several different underlying myocardial substrates. Despite this variability, the surface 12-lead electrocardiograph (ECG) has proven to be a robust and reproducible initial mapping tool that can provide useful information in localizing the origin of both focal and reentrant forms of VT. The second part of this review series will look at the use of the ECG in mapping the various forms of VT encountered in clinical practice. PMID- 19302479 TI - Catheter ablation of idiopathic ventricular tachycardia arising from the aortic root. AB - Most idiopathic ventricular tachycardia (VT) arises from the area of the outflow tracts, and the most common left ventricular site is the aortic root, usually from the right and left sinuses of Valsalva. This site of origin is suggested by specific patterns on the electrocardiogram. Activation mapping and pace mapping are both useful strategies, and their relative benefits and limitations need to be appreciated. The mapping strategy for a VT of suspected aortic root origin is based on the consideration that multiple chambers may need to be mapped, and the temptation to ablate at suboptimal sites based on the justification that it was easy to get to should be resisted. The entire surface of each cusp needs to be sampled, as distinct activation times and pacemaps are obtained at each site. Standard radiofrequency energy is typically adequate and the precision of mapping rather than the amount of tissue ablated is tantamount to success. In my opinion, the indications for ablation of aortic root VT are similar to those for other idiopathic VT. Although I offer patients both pharmacologic and nonpharmacologic options, I feel that ablation is first line therapy for patients with sufficient symptoms to warrant therapy. PMID- 19302480 TI - Acute hemodynamic effects of atrial pacing with cardiac resynchronization therapy. AB - BACKGROUND: Chronotropic incompetence is common among patients with advanced heart failure (HF), thus atrial pacing (AP) is frequently utilized in this population. The hemodynamic effects of AP during cardiac resynchronization therapy (CRT) have not been well studied. OBJECTIVE: The purpose of this study was to compare the acute hemodynamic response during CRT of AP with that during atrial sensing (AS). METHODS: This study included 26 patients undergoing CRT. At implant, invasive left ventricular (LV) dP/dt was measured by a micromanometer catheter during biventricular pacing in AS and AP modes at 5 different atrioventricluar delays (AVD), tested in randomized order. Postimplant, echocardiography was performed to obtain aortic and mitral flow velocity integrals at baseline (no CRT) and during CRT. RESULTS: Compared with intrinsic rhythm, CRT increased LV dP/dt by 11 +/- 11% during AS (heart rate: 74 +/- 13 bpm) and by 17 +/- 11% during AP (heart rate: 86 +/- 12 bpm, P < 0.001). The AVD associated with maximal hemodynamic response (AVD(max)) during AP was 72 +/- 40 ms longer than during AS. However, aortic and mitral flow velocity integrals decreased by 15-20% during AP. The aortic and mitral flow velocities at AVD(max) for LV dP/dt(max) were highly correlated with their maximum values (r > 0.98). CONCLUSION: AP increases LV dP/dt during CRT, but requires a substantially longer AV delay. However, AP results in modest reductions of LV filling and stoke volume. Further studies are needed to assess the long-term impact of AP on HF functional status and LV remodeling. PMID- 19302481 TI - Incessant interfascicular reentrant ventricular tachycardia as a result of catheter ablation of the right bundle branch: case report and review of the literature. AB - A 72-year-old woman developed incessant interfascicular (IF) ventricular tachycardia immediately after successful right bundle branch (RBB) catheter ablation for the treatment of sustained bundle branch reentrant tachycardia. Catheter ablation of the left bundle branch and the left anterior fascicle was successful in eliminating the tachycardia (in 2 different sessions). This report discusses the direct link between the creation of an RBB block and the development of IF tachycardia, in our case, and in prior cases of IF reentry reported in the literature. PMID- 19302482 TI - Protection of midbrain dopaminergic neurons by the end-product of purine metabolism uric acid: potentiation by low-level depolarization. AB - High plasma levels of the end product of purine metabolism uric acid (UA) predict a reduced risk of developing Parkinson's disease suggesting that UA may operate as a protective factor for midbrain dopaminergic neurons. Consistent with this view, UA exerted partial but long-term protection in a culture model in which these neurons die spontaneously. The rescued neurons were functional as they accumulated dopamine, efficiently. The use of the fluorescent probe dihydrorhodamine-123 revealed that UA operated by an antioxidant mechanism. The iron chelating agent desferrioxamine, the H(2)O(2) scavenger enzyme catalase and the inhibitor of lipid peroxidation Trolox mimicked the effects of UA, suggesting that UA neutralized reactive oxygen species produced via a Fenton-type chemical reaction. UA was, however, not significantly accumulated into neurons, which indicates that the antioxidant effect occurred probably extracellularly. Structure - activity relationships among purine derivatives revealed that the antioxidant properties of UA resulted from the presence of a 8-one substituent in its chemical structure. Of interest, the stimulation of L-type Ca(2+) channels by high K(+)-induced depolarization and the ensuing activation of extracellular signal-regulated kinases 1/2 strongly improved the neuroprotective effect of UA whereas the depolarizing signal alone had no effect. In summary, our data indicate that UA may interfere directly with the disease's pathomechanism. PMID- 19302483 TI - Regulation of dendritic spine morphology by SPIN90, a novel Shank binding partner. AB - Dendritic spines are highly specialized actin-rich structures on which the majority of excitatory synapses are formed in the mammalian CNS. SPIN90 is an actin-binding protein known to be highly enriched in postsynaptic densities (PSDs), though little is known about its function there. Here, we show that SPIN90 is a novel binding partner for Shank proteins in the PSD. SPIN90 and Shank co-immunoprecipitate from brain lysates and co-localize in postsynaptic dendrites and act synergistically to mediate spine maturation and spine head enlargement. At the same time, SPIN90 causes accumulation of Shank and PSD-95 within dendritic spines. In addition, we found that the protein composition of PSDs in SPIN90 knockout mice is altered as is the actin cytoskeleton of cultured hippocampal SPIN90 knockout neurons. Taken together, these findings demonstrate that SPIN90 is a Shank1b binding partner and a key contributor to the regulation of dendritic spine morphogenesis and brain function. PMID- 19302485 TI - Abnormal production of macrophage inflammatory protein-1alpha by microglial cell lines derived from neonatal brains of Sandhoff disease model mice. AB - Sandhoff disease (SD) is a lysosomal beta-hexosaminidase deficiency involving excessive accumulation of undegraded substrates, including terminal N acetylglucosamine-oligosaccharides and GM2 ganglioside, and progressive neurodegeneration. Our previous study demonstrated remarkable induction of macrophage inflammatory factor-1alpha (MIP-1alpha) in microglia in the brains of SD model mice as a putative pathogenic factor for SD via microglia-mediated neuroinflammation. In this study, we established microglial cell lines (WT- and SD-Mg) from wild-type and SD mice, and first demonstrated the enhanced production of MIP-1alpha in SD-Mg. Inhibitors of protein kinase C (PKC) and Akt reduced the production of MIP-1alpha by SD-Mg. Elevated activation of Akt and partial translocation of PKC isozymes (alpha, betaI, betaII, and delta) from the cytoplasm to the membrane in SD-Mg were also revealed by means of immunoblotting. Furthermore, it was demonstrated that intracellular extracellular signal regulated kinase, c-Jun N-terminal kinase, and phospholipase C (PLC), but not phosphoinositide 3-kinase, should contribute to the induction of MIP-1alpha in SD Mg, and that PLC could independently regulate the activation of both PKC and Akt. We proposed here that the deregulated activation of PLC should cause the enhanced MIP-1alpha production via plural signaling pathways mediated by PKC and Akt, followed by extracellular signal-regulated kinase and c-Jun N-terminal kinase, in SD-Mg. PMID- 19302484 TI - A role for TREM2 ligands in the phagocytosis of apoptotic neuronal cells by microglia. AB - Following neuronal injury, microglia initiate repair by phagocytosing dead neurons without eliciting inflammation. Prior evidence indicates triggering receptor expressed by myeloid cells-2 (TREM2) promotes phagocytosis and retards inflammation. However, evidence that microglia and neurons directly interact through TREM2 to orchestrate microglial function is lacking. We here demonstrate that TREM2 interacts with endogenous ligands on neurons. Staining with TREM2-Fc identified TREM2 ligands (TREM2-L) on Neuro2A cells and on cultured cortical and dopamine neurons. Apoptosis greatly increased the expression of TREM2-L. Furthermore, apoptotic neurons stimulated TREM2 signaling, and an anti-TREM2 mAb blocked stimulation. To examine the interaction between TREM2 and TREM2-L in phagocytosis, we studied BV2 microglial cells and their engulfment of apoptotic Neuro2A. One of our anti-TREM2 mAb, but not others, reduced engulfment, suggesting the presence of a functional site on TREM2 interacting with neurons. Further, Chinese hamster ovary cells transfected with TREM2 conferred phagocytic activity of neuronal cells demonstrating that TREM2 is both required and sufficient for competent uptake of apoptotic neuronal cells. Finally, while TREM2 L are expressed on neurons, TREM2 is not; in the brain, it is found on microglia. TREM2 and TREM2-L form a receptor-ligand pair connecting microglia with apoptotic neurons, directing removal of damaged cells to allow repair. PMID- 19302486 TI - Transcriptional regulation of metabolic pathways, alternative respiration and enterotoxin genes in anaerobic growth of Bacillus cereus ATCC 14579. AB - AIMS: To assess genes specifically activated during anaerobic growth that are involved in metabolism and pathogenesis of the foodborne pathogen Bacillus cereus. METHODS AND RESULTS: Growth under anaerobic conditions in Brain Heart Infusion (BHI) broth revealed a reduced growth rate and lower yield as compared to growth under aerobic conditions. Subsequently, comparative transcriptome analysis showed specific genes induced under anaerobic conditions. These included novel genes identified for anaerobic growth of B. cereus, encoding metabolic pathways, such as the arginine deiminase pathway (ArcABDC), formate dehydrogenase (FdhF) and pyruvate formate lyase (Pfl), and alternative respiratory proteins, such as arsenate reductases. Notably, haemolytic enzyme encoding genes were induced during anaerobic growth, and enterotoxin genes were induced in high cell density transition and stationary phases of aerobic cultures. CONCLUSIONS: These data point to induction of stress adaptation and pathogenicity factors and rearrangements of expression of metabolic pathways in response to oxygen limitations in B. cereus. SIGNIFICANCE AND IMPACT OF THE STUDY: The reported changes in gene expression show that the foodborne pathogen B. cereus can adjust to anaerobic conditions, such as encountered in the human GI-tract. PMID- 19302487 TI - FDB2 encodes a member of the arylamine N-acetyltransferase family and is necessary for biotransformation of benzoxazolinones by Fusarium verticillioides. AB - AIMS: To clone and characterize genes from the mycotoxigenic fungus, Fusarium verticillioides, which are associated with its ability to biotransform allelopathic benzoxazolinones produced by maize, wheat, and rye. METHODS AND RESULTS: Suppression subtractive hybridization identified F. verticillioides genes up-regulated in response to 2-benzoxazolinone (BOA), including a cluster of genes along chromosome 3. One of these genes, putatively encoding an arylamine N acetyltransferase (NAT), was highly represented in the subtracted library and was of particular interest since previous analyses identified the FDB2 locus as possibly encoding transferase activity. The gene was subcloned and complemented a natural fdb2 mutant. Conversely, disruption of the gene eliminated the ability of F. verticillioides to metabolize BOA. Other genes in the cluster also were assessed using a complementation assay. Metabolic profiles of fdb2 mutants suggest that minor acylation activity occurred independently of the NAT activity encoded by FDB2. CONCLUSIONS: The previously defined FDB2 locus was functionally associated with the gene encoding putative NAT activity, and the FDB2 gene was essential for biotransformation of BOA. The flanking gene FDB3 encodes a putative Zn(II)2Cys6 transcription factor and contributes to efficient BOA biotransformation but was not essential. SIGNIFICANCE AND IMPACT OF THE STUDY: Biotransformation of benzoxazolinones by F. verticillioides may enhance its ecological fitness in maize field environments and our results provide greater understanding of the genes that modulate the biotransformation process. Additionally, this is the first homologue of the NAT gene family to be characterized in a filamentous fungus. PMID- 19302488 TI - Populations of heavy fuel oil-degrading marine microbial community in presence of oil sorbent materials. AB - AIMS: To investigate the feasibility of applying sorbent material X-Oil in marine oil spill mitigation and to survey the interactions of oil, bacteria and sorbent. METHODS AND RESULTS: In a series of microcosms, 25 different treatments including nutrient amendment, bioaugmentation with Alcanivorax borkumensis and application of sorbent were tested. Microbial community dynamics were analysed by DNA fingerprinting methods, RISA and DGGE. Results of this study showed that the microbial communities in microcosms with highly active biodegradation were strongly selected in favour of A. borkumensis. Oxygen consumption measurements in microcosms and gas chromatography of oil samples indicated the fast and intense depletion of linear alkanes as well as high oxygen consumption within 1 week followed by consequent slower degradation of branched and polyaromatic hydrocarbons. CONCLUSION: Under given conditions, A. borkumensis was an essential organism for biodegradation, dominating the biofilm microbial community formation and was the reason of emulsification. SIGNIFICANCE AND IMPACT OF THE STUDY: This study strongly emphasizes the pivotal importance of A. borkumensis as an essential organism in the initial steps of marine hydrocarbon degradation. Interaction with the sorbent material X-Oil proved to be neutral to beneficial for biodegradation and also promoted the growth of yet unknown micro-organisms. PMID- 19302489 TI - Functional, genetic and chemical characterization of biosurfactants produced by plant growth-promoting Pseudomonas putida 267. AB - AIMS: Plant growth-promoting Pseudomonas putida strain 267, originally isolated from the rhizosphere of black pepper, produces biosurfactants that cause lysis of zoospores of the oomycete pathogen Phytophthora capsici. The biosurfactants were characterized, the biosynthesis gene(s) partially identified, and their role in control of Phytophthora damping-off of cucumber evaluated. METHODS AND RESULTS: The biosurfactants were shown to lyse zoospores of Phy. capsici and inhibit growth of the fungal pathogens Botrytis cinerea and Rhizoctonia solani. In vitro assays further showed that the biosurfactants of strain 267 are essential in swarming motility and biofilm formation. In spite of the zoosporicidal activity, the biosurfactants did not play a significant role in control of Phytophthora damping-off of cucumber, since both wild type strain 267 and its biosurfactant deficient mutant were equally effective, and addition of the biosurfactants did not provide control. Genetic characterization revealed that surfactant biosynthesis in strain 267 is governed by homologues of PsoA and PsoB, two nonribosomal peptide synthetases involved in production of the cyclic lipopeptides (CLPs) putisolvin I and II. The structural relatedness of the biosurfactants of strain 267 to putisolvins I and II was supported by LC-MS and MS-MS analyses. CONCLUSIONS: The biosurfactants produced by Ps. putida 267 were identified as putisolvin-like CLPs; they are essential in swarming motility and biofilm formation, and have zoosporicidal and antifungal activities. Strain 267 provides excellent biocontrol activity against Phytophthora damping-off of cucumber, but the lipopeptide surfactants are not involved in disease suppression. SIGNIFICANCE AND IMPACT OF THE STUDY: Pseudomonas putida 267 suppresses Phy. capsici damping-off of cucumber and provides a potential supplementary strategy to control this economically important oomycete pathogen. The putisolvin-like biosurfactants exhibit zoosporicidal and antifungal activities, yet they do not contribute to biocontrol of Phy. capsici and colonization of cucumber roots by Ps. putida 267. These results suggest that Ps. putida 267 employs other, yet uncharacterized, mechanisms to suppress Phy. capsici. PMID- 19302490 TI - Investigation of bacteria with polyketide synthase genes and antimicrobial activity isolated from South China Sea sponges. AB - AIMS: To obtain bacteria with PKS (polyketide synthase) genes and antimicrobial activity from sponges. METHODS AND RESULTS: Eighteen bacteria with KS (ketosynthase) genes were identified by polymerase chain reaction (PCR) screening of 98 isolates from South China Sea sponges, Stelletta tenuis, Halichondria rugosa, Dysidea avara and Craniella australiensis. 16S rRNA gene-based Blast analysis indicated that 15 isolates belonged to the phylum Firmicutes, among which 14 isolates were closely related to genus Bacillus, and 1 to Staphylococcus lentus. Two isolates were identified as actinomycetes, and one as Alcaligenes sp. in the phylum Proteobacteria. The 18 KS domains belong to trans-AT type I PKS and match PKS of marine bacterial symbionts. The 18 bacteria exhibited broad-spectrum antimicrobial activities against fungi, gram-positive and gram-negative bacteria. A 21.8-kb PKS gene cluster fragment containing five modules was isolated from the Staphylococcus lentus isolate A75 by screening of a fosmid library. CONCLUSIONS: The PKS gene diversity and different antimicrobial spectra indicate the potential of bacteria associated with South China Sea sponges for diverse polyketide production. SIGNIFICANCE AND IMPACT OF THE STUDY: Combined with bioactivity assay the PKS gene-based approach can be applied to efficient screening of strains of pharmaceutical value and the prediction of related compounds. PMID- 19302491 TI - Integron-containing bacteria in faeces of cattle from different production systems at slaughter. AB - AIMS: To determine the prevalence and characteristics of integron-containing bacteria in faeces of cattle from grass-fed, lot-fed, or organically produced cattle. METHODS AND RESULTS: Faecal samples from grass-fed (n = 125), lot-fed (n = 125) and organic (n = 135) cattle were tested for the presence of class 1 and class 2 integrons by using PCR and colony hybridisation. The prevalence of class 1 and class 2 integrase were higher in lot-fed cattle (71% and 62%) than grass fed cattle (52% and 30%) which in turn were higher than organic cattle (25% and 11%). Isolation rates of integron-containing bacteria were reflective of PCR prevalence results. CONCLUSIONS: The antimicrobial resistance genes harboured by the integrons differed little across the three systems and were typically to antimicrobials that would rarely be used therapeutically or for growth promotion purposes. The differences in prevalence observed between the systems may be a function of the intensiveness of each system. SIGNIFICANCE AND IMPACT OF THE STUDY: Integron-containing bacteria may be present in all cattle production systems regardless of the amount of antimicrobial use and confirms that the prudent use of antimicrobials is required so that the development of integrons harbouring genes significant to human medicine is avoided. PMID- 19302492 TI - Antibacterial activity of fresh and processed red muscadine juice and the role of their polar compounds on Escherichia coli O157:H7. AB - AIMS: The objectives of this research were to show the anti-Escherichia coli O157:H7 effect of fresh (FRMJ) and processed red muscadine (Vitis rotundifolia) juice (PRMJ) and to discern the active compounds responsible for anti-E. coli O157:H7. METHODS AND RESULTS: Polar and phenolic compounds of FRMJ and PRMJ were analysed by high-performance liquid chromatography. Antibacterial activity of FRMJ, PRMJ, their polar and polyphenol fractions, individual synthetic acids and their mixture with or without sugars were investigated on E. coli O157:H7. FRMJ and PRMJ inactivated (P < or = 0.05) 5-log cocktail cells of E. coli O157:H7 within 4 h at 37 degrees C. Polar fractions that contained malic, tartaric and tannic acids showed strong antimicrobial activity (P < or = 0.05) against E. coli O157:H7. Tannic acid among the synthetic acids showed the highest antimicrobial activity against E. coli O157:H7. CONCLUSIONS: FRMJ, PRMJ and their polar compounds showed strong anti-E. coli O157:H7 activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Earlier findings have failed to show any anti-E. coli O157:H7 effect of grape juice without adding preservatives. Our findings show that red muscadine juice has natural antibacterial substances and suggest that these can be used as active antimicrobial ingredients against E. coli O157:H7 in nonalcoholic beverages. PMID- 19302493 TI - Nonisothermal heat resistance determinations with the thermoresistometer Mastia. AB - AIMS: To design and build a thermoresistometer, named Mastia, which could perform isothermal and nonisothermal experiments. METHODS AND RESULTS: In order to evaluate the thermoresistometer, the heat resistance of Escherichia coli vegetative cells and Alicyclobacillus acidoterrestris spores was explored. Isothermal heat resistance of E. coli was characterized by D(60 degrees C) = 0.38 min and z = 4.7 degrees C in pH 7 buffer. When the vegetative cells were exposed to nonisothermal conditions, their heat resistance was largely increased at slow heating and fast cooling rates. Isothermal heat resistance of A. acidoterrestris was characterized by D(95 degrees C) = 7.4 min and z = 9.5 degrees C in orange juice. Under nonisothermal conditions, inactivation was reasonably well predicted from isothermal data. CONCLUSIONS: The thermoresistometer Mastia is a very suitable instrument to get heat resistance data of micro-organisms under isothermal and nonisothermal treatments. SIGNIFICANCE AND IMPACT OF THE STUDY: The thermoresistometer Mastia can be a helpful tool for food processors in order to estimate the level of safety of the treatments they apply. PMID- 19302494 TI - Infection biology and defence responses in sorghum against Colletotrichum sublineolum. AB - AIMS: To investigate the infection biology of Colletotrichum sublineolum (isolate CP2126) and defence responses in leaves of resistant (SC146), intermediately resistant (SC326) and susceptible (BTx623) sorghum genotypes. METHODS AND RESULTS: Infection biology and defence responses were studied quantitatively by light microscopy, H(2)O(2) accumulation by DAB staining and HRGP accumulation by immunological methods. Inhibition of conidial germination and appressorium formation may represent prepenetration defence responses on the leaf surface. Inducible defence responses in the resistant genotypes included decreases in formation of appressoria as well as accumulation of H(2)O(2), HRGPs and phytoalexins. Concomitant with these inducible responses, fungal growth was stopped during or just after penetration in genotypes SC146 and SC326. High levels of H(2)O(2) accumulating at late infection stages (5 days after inoculation) in the susceptible genotype BTx623 correlated with necrosis and tissue degeneration. CONCLUSIONS: The early accumulation of H(2)O(2) and HRGPs indicates roles in defence whereas the late accumulation in genotype BTx623 correlated with successful pathogenesis. SIGNIFICANCE AND IMPACT OF THE STUDY: The fact that there is a significant correlation between induced accumulation of H(2)O(2), papilla formation and cell wall cross-linking, as evidenced by HRGP accumulation, and cessation of pathogen growth in resistant genotypes may help exploit host resistance in sorghum. PMID- 19302495 TI - Characterization and purification of a new bacteriocin with a broad inhibitory spectrum produced by Lactobacillus plantarum lp 31 strain isolated from dry fermented sausage. AB - AIMS: Characterization and purification of a new bacteriocin produced by Lactobacillus plantarum LP 31 strain, isolated from Argentinian dry-fermented sausage. METHODS AND RESULTS: Lactobacillus plantarum LP 31 strain produces an antimicrobial compound that inhibits the growth of food-borne pathogenic bacteria. It was inactivated by proteolytic enzymes, was stable to heat and catalase and exhibited maximum activity in the pH range from 5.0 to 6.0. Consequently, it was characterized as a bacteriocin. It was purified by RP (reverse-phase) solid-phase extraction, gel filtration chromatography and RP HPLC. Plantaricin produced by Lact. plantarum LP 31 is a peptide with a molecular weight of 1558.85 Da as determined by Maldi-Tof mass spectrometry and contains 14 amino acid residues. It was shown to have a bactericidal effect against Pseudomonas sp., Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes. CONCLUSIONS: The bacteriocin produced by Lact. plantarum LP 31 may be considered as a new plantaricin according to its low molecular weight and particular amino acid composition. SIGNIFICANCE AND IMPACT OF THE STUDY: In view of the interesting inhibitory spectrum of this bacteriocin and because of its good technological properties (resistance to heat and activity at acidic pH), this bacteriocin has potential applications as a biopreservative to prevent the growth of food-borne pathogens and food spoilage bacteria in certain food products. PMID- 19302496 TI - Lower urinary tract symptoms: new perspectives on prevalence, burden and comorbidities. Introduction. PMID- 19302497 TI - The burden of lower urinary tract symptoms: evaluating the effect of LUTS on health-related quality of life, anxiety and depression: EpiLUTS. AB - OBJECTIVE: To evaluate the impact of lower urinary tract symptoms (LUTS) on urinary-specific health-related quality of life (HRQL), generic health indices, depression and anxiety in a population-representative sample of men and women, as research has linked LUTS with reduced HRQL and depression, but little is known about the effects of individual LUTS on HRQL, depression and anxiety. SUBJECTS AND METHODS: A cross-sectional population-representative survey was conducted via the Internet in the USA, the UK and Sweden. Participants rated the frequency and symptom-specific bother of individual LUTS and condition-specific HRQL, generic health status, anxiety and depression. Descriptive statistics were used to evaluate outcome differences by International Continence Society LUTS subgroups; logistic regressions were used to determine associations of LUTS and perception of bladder problems, anxiety and depression. RESULTS: The overall survey response rate was 59.2%; 30 000 subjects (14 139 men and 15 861 women) participated. Men and women with LUTS in the all LUTS subgroup (storage, voiding and postmicturition) reported the lowest levels of HRQL and highest levels of anxiety and depression, with 35.9% of men and 53.3% of women meeting self-reported screening criteria for clinical anxiety (Hospital Anxiety and Depression Scale, HADS, Anxiety > or =8), and 29.8% of men and 37.6% of women meeting self-reported criteria for clinical depression (HADS Depression > or =8). In both men and women, storage symptoms were significantly associated with greater perceived bladder impact, whereas voiding symptoms were not. Significant predictors of anxiety included nocturia, urgency, stress urinary incontinence, leaking during sexual activity, weak stream and split stream in women; and nocturia, urgency, incomplete emptying and bladder pain in men. For depression, weak stream, urgency and stress urinary incontinence were significant for women, and perceived frequency and incomplete emptying were significant for men. CONCLUSION: The negative effect of LUTS is apparent across several domains of HRQL and on overall perception of bladder problems, general health status and mental health. The high level of psychiatric morbidity in patients with multiple LUTS has important implications for treatment and highlights the need for further research to pinpoint specific mechanisms underlying this association. PMID- 19302498 TI - The overlap of storage, voiding and postmicturition symptoms and implications for treatment seeking in the USA, UK and Sweden: EpiLUTS. AB - OBJECTIVE: To assess the (i) the overlap between voiding, storage, and postmicturition symptoms; and (ii) the relative effect of bother and implications for treatment seeking within these symptom groups, using data from the EpiLUTS study. SUBJECTS AND METHODS: This cross-sectional population-representative survey was conducted via the Internet in the USA, the UK and Sweden. Participants were asked to rate the frequency and symptom-specific bother of individual LUTS. Descriptive statistics were used to examine differences in International Continence Society LUTS subgroups. Logistc regressions were used with treatment seeking as the dependent variable and the bother of individual symptoms as predictors. RESULTS: The survey response rate was 59%. The sample included 30,000 participants (14,139 men and 15,861 women); 71% of men and 75% of women reported at least one LUTS, and about half reported LUTS from more than one symptom group. Rates of bother were greatest for those who reported multiple storage, voiding and postmicturition LUTS (men 83%, women 89%). Less than a third of participants with LUTS from all three groups reported seeking treatment. Consistent correlates of treatment seeking across genders included bother due to weak stream, incomplete emptying, perceived daytime frequency, nocturia and urgency. There were also significant associations for several types of incontinence, most commonly stress incontinence in women and leaking during sexual activity in men. Despite high rates of symptom overlap and symptom-specific bother, few participants sought treatment for LUTS. CONCLUSION: Common conditions such as BPH and OAB are treatable, and clinicians should proactively ask patients about urinary symptoms. Given the many types of LUTS that patients experience, it is imperative that clinicians assess all LUTS to ensure that appropriate treatments are prescribed. PMID- 19302499 TI - Risk factors and comorbid conditions associated with lower urinary tract symptoms: EpiLUTS. AB - OBJECTIVE: To explore the risk factors and comorbid conditions associated with subgroups of lower urinary tract symptoms (LUTS) in men and women aged > or =40 years in three countries, using data from the EpiLUTS study, as LUTS are common amongst men and women and increase in prevalence with age. SUBJECTS AND METHODS: This cross-sectional, population-representative survey was conducted via the Internet in the USA, the UK and Sweden. Participants were asked to rate how often they experienced individual LUTS during the past 4 weeks on a 5-point Likert scale. Eight LUTS subgroups were created. Descriptive statistics and logistic regressions within each LUTS subgroup were used to assess the data. RESULTS: The survey response rate was 59%. The final sample was 30,000 (men and women). The voiding + storage + postmicturition (VSPM) group reported the highest rates of comorbid conditions for both men and women, and the fewest were reported in the no/minimal LUTS and the postmicturition-only groups. Increasing age was associated with increasing LUTS in men, but not in women. Comorbid conditions significantly associated with the VSPM group were arthritis, asthma, chronic anxiety, depression, diabetes (men only), heart disease, irritable bowel syndrome, neurological conditions, recurrent urinary tract infection, and sleep disorders. Risk factors, such as body mass index, exercise level and smoking, played less of a role, except for childhood nocturnal enuresis, which was significantly associated with most LUTS subgroups. CONCLUSION: In this large population study, many comorbid conditions and risk factors were significantly associated with LUTS among both men and women. Further longitudinal investigations of the associations noted here would help physicians to understand the pathophysiology of LUTS and comorbid conditions, and provide clinical guidelines for patient management of comorbid conditions sharing common pathophysiological pathways. PMID- 19302500 TI - The impact of lower urinary tract symptoms on male sexual health: EpiLUTS. AB - OBJECTIVE: To evaluate the association between International Continence Society categories of lower urinary tract symptoms (LUTS; storage, voiding, and postmicturition) and individual LUTS (associated with decreased sexual activity and sexual satisfaction in men) with erectile dysfunction (ED), ejaculatory dysfunction (EjD) and premature ejaculation (PE). SUBJECTS AND METHODS: The impact of LUTS on men's sexual health was captured as part of a cross-sectional epidemiological study to assess the prevalence LUTS among men and women aged > or =40 years in the USA, the UK and Sweden. RESULTS: The analysis included 11 834 men with a mean age of 56.1 years, 71% of whom reported being currently sexually active. The primary reason for not being sexually active was no partner (35%), followed by personal health (23%) and no desire (23%). Of the men, 26% had mild to severe ED, 7% had EjD, and 16% PE. Men with multiple LUTS had more severe ED and more frequent EjD and PE. Logistic regression analysis showed that greater age, hypertension, diabetes, depression, urgency with fear of leaking, and leaking during sexual activity were significantly associated with ED. The results were similar in the logistic regression analysis for EjD, whereas being younger and the absence of prostatitis were significantly associated with PE, as were the presence of terminal dribble, incomplete emptying, and split stream. CONCLUSION: LUTS are associated with common sexual dysfunctions in men. The results of this study highlight the importance of assessing the sexual health of men presenting with LUTS. PMID- 19302501 TI - Lower urinary tract symptoms and sexual health: the role of gender, lifestyle and medical comorbidities. AB - OBJECTIVE: To investigate the association between urological symptoms and self reported measures of sexual activity, desire and function in large representative samples of men and women in the Boston Area Community Health (BACH) survey. SUBJECTS AND METHODS: The BACH survey is a racially and ethnically diverse random sample of 5503 community-dwelling residents aged 30-79 years, of Boston, MA, USA. Urological symptoms and sexual function were assessed in men and women at baseline using validated self-report measures, i.e. the International Prostate Symptom Score (IPSS), International Index of Erectile Function, and Female Sexual Function Index. Bivariate and multivariate analyses were conducted to assess the degree of association of sexual function measures with LUTS and other common urological problems in men and women in the BACH sample, controlling for the effects of age, medical comorbidities, and lifestyle factors. RESULTS: Urological symptoms were associated with a significant decrease in sexual activity and function in both men and women. Women were more likely to report low sexual desire than men. Low sexual desire was associated with depression, nocturia and prostatitis in men. Erectile dysfunction in men was significantly associated with LUTS, nocturia and prostatitis in bivariate associations, and with prostatitis in multivariate analyses, controlling for the effects of diabetes and other comorbidities. In the multivariate analysis in women, sexual dysfunction was primarily associated with depression and inversely with alcohol use. CONCLUSION: Sexual activity and function were diminished in both men and women with urological symptoms, although women reported more sexual problems overall than men, and the profile of risk factors and comorbidities was different across genders. PMID- 19302502 TI - Implications of recent epidemiology studies for the clinical management of lower urinary tract symptoms. AB - Lower urinary tract symptoms (LUTS) include storage, voiding, and postmicturition symptoms, and occur commonly in both men and women. Findings from two recent epidemiological studies, the Epidemiology of LUTS study and the Boston Area Community Health survey, further extend the understanding of the prevalence of individual LUTS, the overlap of LUTS in men and women, the associations of LUTS with other comorbid conditions, the impact of LUTS on health-related quality of life (HRQL), and the relationships between frequency and bother of LUTS and treatment-seeking behaviour. Examining the clinical implications of these findings might provide directions to physicians for managing their patients with LUTS. For example, common findings of separate patient groups spanning a spectrum from those with typically one urinary symptom of mild to moderate severity to those with multiple more severe LUTS and frequent comorbidities might further encourage the diagnosis and treatment of comorbid conditions as a standard part of the management of patients with LUTS. Likewise, understanding that the impact of LUTS on HRQL and the degree of bother, rather than the frequency of LUTS, are significant drivers for treatment seeking might aid in assisting patients to make decisions about treatment. PMID- 19302503 TI - Indoor thermal factors and symptoms in office workers: findings from the US EPA BASE study. AB - Some prior research in office buildings has associated higher indoor temperatures even within the recommended thermal comfort range with increased worker symptoms. We reexamined this relationship in data from 95 office buildings in the US Environmental Protection Agency's Building Assessment Survey and Evaluation Study. We investigated relationships between building-related symptoms and thermal metrics constructed from real-time measurements. We estimated odds ratios and 95% confidence intervals in adjusted logistic regression models with general estimating equations, overall and by season. Winter indoor temperatures spanned the recommended winter comfort range; summer temperatures were mostly colder than the recommended summer range. Increasing indoor temperatures, overall, were associated with increases in few symptoms. Higher winter indoor temperatures, however, were associated with increases in all symptoms analyzed. Higher summer temperatures, above 23 degrees C, were associated with decreases in most symptoms. Humidity ratio, a metric of absolute humidity, showed few clear associations. Thus, increased symptoms with higher temperatures within the thermal comfort range were found only in winter. In summer, buildings were overcooled, and only the higher observed temperatures were within the comfort range; these were associated with decreased symptoms. Confirmation of these findings would suggest that thermal management guidelines consider health effects as well as comfort, and that less conditioning of buildings in both winter and summer may have unexpected health benefits. PRACTICAL IMPLICATIONS: In winter, higher temperatures within the thermal comfort range are common in US office buildings and may be associated with increased symptoms. In summer, temperatures below the thermal comfort range are common and may be associated with increased symptoms. Results from this large study thus suggest that in US office buildings, less winter heating (in buildings that are in heating mode) and less summer cooling may reduce acute symptoms while providing substantial energy conservation benefits, with no expected thermal comfort penalty and, in summer, even thermal comfort benefits. If confirmed, this would be welcome news. PMID- 19302504 TI - Detection and molecular characterization of filamentous actinobacteria and thermoactinomycetes present in water-damaged building materials. AB - In this study the dominant filamentous actinobacteria occurring in water-damaged building materials were detected by culture and characterized by automated ribotyping and 16S rRNA gene sequencing. Fifty-two samples were taken from 20 water-damaged houses in four different countries. A total of 122 bacterial isolates were analyzed. Actinobacteria or thermoactinomycetes were present in 48% of the samples. The dominant genus was Streptomyces (58% of isolates), followed by Thermoactinomyces (23%), Laceyella (14%), Nocardiopsis (3%), Pseudonocardia (1%) and Saccharomonospora (1%). The most frequently detected species was the thermophilic Thermoactinomyces vulgaris (14 samples/4 countries). The most common streptomycetes were closely related to the heterogeneous species Streptomyces microflavus (7/2) or Streptomyces griseus (6/2). Automated ribotyping was a rapid tool for reliable characterization of these isolates. The spores of thermoactinomycetes and toxic substances of Nocardiopsis species and S. griseus may constitute a risk for human health. PRACTICAL IMPLICATIONS: Harmful microbes in indoor environments are a cause of public concern. To develop rapid and simple to-use molecular biological methods to detect the presence of harmful actinobacterial species in water-damaged buildings more information about their occurrence in those materials is needed, which this study provides. PMID- 19302505 TI - Regional differences in residential environments and the association of dwellings and residential factors with the sick house syndrome: a nationwide cross sectional questionnaire study in Japan. AB - This study was conducted to clarify regional differences in residential factors and the association of those factors with dwellings having sick house syndrome (SHS) problems. The survey was conducted in six areas of northern and southern Japan. In terms of regional differences, dampness was not as severe in the dwellings in Sapporo as compared with that in areas in the south. SHS was defined using five categories of nasal, throat and respiratory, skin and general symptoms, which appeared frequently or not frequently and improved upon leaving the home. The dampness index was estimated by the sum of the presence of several indicators: condensation on the window panes and/or wall, visible mold growth, moldy odor, slow-drying wet towels in the bathroom, and water leakage. The dwellings where inhabitants showed any symptoms of SHS comprised 3.7% of all surveyed dwellings. We found significant associations between SHS and dampness index, odors, and stuffiness of the air. For dampness, the adjusted odds ratio (OR) increased with increased dampness index, adjusting for the age of the house, pets indoors, stuffiness of the air, and odors. These results showed an increased risk when several dampness indicators appeared simultaneously. PRACTICAL IMPLICATIONS: To evaluate the associations of residential environments and Sick House Syndrome (SHS), this cross-sectional questionnaire study was conducted on 2297 dwellings in six areas in Japan from 2003 to 2004. The dwellings where inhabitants showed any of nasal, throat and respiratory, skin and general symptoms comprised 3.7% of all surveyed dwellings, and an increased risk for SHS was found when several dampness indicators, 'condensation', 'visible mold growth', 'moldy odor', 'slow drying wet towels in the bathroom' and 'water leakage', appeared simultaneously. PMID- 19302506 TI - Radical retropubic prostatectomy with running vesicourethral anastomosis and early catheter removal: our experience. AB - OBJECTIVES: To assess the outcomes of patients undergoing radical retropubic prostatectomy (RRP) with a running vesicourethral anastomosis and catheter removal on postoperative day 3 or 5. METHODS: From February 2006 through December 2007, 55 patients underwent RRP at our institution. All procedures were performed by a single surgeon using a running suture for the vesicourethral anastomosis. A cystogram was carried out before catheter removal in all patients. The initial 23 of 55 patients (Group 1; n = 23) had the cystogram on postoperative day 5, the other 32 patients (Group 2; n = 32) had the cystogram on postoperative day 3. Removal of the catheter was only carried out if there was no anastomotic extravasation. RESULTS: The success rate of catheter removal in group 1 and 2 was 100% and 96.9%, respectively. Overall continence rates were 83.3%, 87% and 90.7% at 24, 48 and 72 h after removal of the catheter, respectively. There was no significant difference in terms of continence rate between groups 1 and 2. None of the patients had acute urinary retention and/or anastomotic stricture after catheter removal. CONCLUSIONS: These findings suggest that an advanced running vesicourethral anastomosis during RRP is technically feasible, allowing safe early catheter removal in most patients. PMID- 19302507 TI - Type-I interferon receptor expression: its circadian rhythm and downregulation after interferon-alpha administration in peripheral blood cells from renal cancer patients. AB - OBJECTIVES: To investigate the regulation of interferon-alpha (IFN-alpha) receptor expression in metastatic renal cell carcinoma (RCC) after IFN-alpha administration. METHODS: Blood sampling was carried out in eight patients with metastatic RCC and six healthy volunteers. Flow-cytometric analysis using a monoclonal antibody against the active subunit of the type-I IFN-alpha receptor (IFNAR2) was carried out to examine the circadian rhythm of IFNAR2 expression in peripheral blood mononuclear cells (PBMC) as well as its downregulation after IFN alpha administration. RESULTS: According to its circadian rhythm IFNAR2 in PBMC had a peak expression at night. Once IFN-alpha is administered, IFNAR2 levels in PBMC showed downregulation within 48 h and recovered within another 48 h. CONCLUSIONS: Our findings might support the establishment of an optimal schedule for IFN-alpha administration. PMID- 19302509 TI - Chronic hypothyroidism only marginally affects adult-type Leydig cell regeneration after EDS administration. AB - Chronic prenatally induced dietary hypothyroidism delays adult-type Leydig cell development, but does not block this process. Using a chemical model to induce hypothyroidism, it was suggested that development of a new population of Leydig cells was completely inhibited following the addition of the cytotoxic compound ethane-1,2-dimethyl sulphonate (EDS). In this study, we used a dietary approach to induce hypothyroidism and reinvestigated the regeneration of the Leydig cell population following EDS administration. Eighty-four day old euthyroid and chronically hypothyroid rats received an injection of EDS and were killed directly before or at regular intervals up to 77 days after EDS. In some control and hypothyroid animals, the first progenitor-type Leydig cells were observed at day 12 after EDS. At day 16, Leydig cell progenitors were present in all rats. The percentage of proliferating Leydig cells peaked in the euthyroid animals at day 21 after EDS. In the hypothyroid testis such a peak was not observed, although the percentage of proliferating regenerating Leydig cells was significantly higher from days 35 to 56 compared with the controls. This suggested that the wave of Leydig cell proliferation was delayed in the hypothyroid animals as compared with the euthyroid controls. On the day of EDS injection, the Leydig/Sertoli cell ratio was 37% lower in the hypothyroid rats compared with the controls. The Leydig/Sertoli cell ratio remained lower in the EDS-treated hypothyroid animals compared with the controls at all time points investigated. At day 77 after EDS, the Leydig cell population had returned to its pre-treatment size in both groups. Plasma testosterone production was reduced to below detectable levels immediately after EDS injection, and started to increase again on day 16, reaching pre-treatment values on day 21 in both groups. Taken together, severely reduced thyroid hormone levels did not block the regeneration of the adult-type Leydig cell population following EDS, as has been suggested previously. PMID- 19302510 TI - The level of polyaromatic hydrocarbons in kajal and surma of major Indian brands. AB - Kajal and surma are eye cosmetics extensively used in Indian subcontinent. Kajal is prepared by burning of vegetable oil and butter oil while surma by grinding of the stones. High performance liquid chromatography and gas chromatography-mass spectrometry instruments were used for quantification and confirmation of 16 polyaromatic hydrocarbons (PAHs). Significant concentration of PAH was found in all the samples examined. The median concentration of PAH ranged from 0.14 (lowest, anthracene) to 31.18 microg g(-1) [dibenz(a,h)anthracene] in kajal sample and from not detectable concentration (naphthalene) to 197.47 microg g(-1) of benzo(a)pyrene in surma sample. Fifteen PAHs were detected in all the samples. Therefore the use of kajal and surma in eye should be strictly restricted. PMID- 19302511 TI - Self-preserving cosmetics. AB - Preservatives are added to products for two reasons: first, to prevent microbial spoilage and therefore to prolong the shelf life of the product; second, to protect the consumer from a potential infection. Although chemical preservatives prevent microbial growth, their safety is questioned by a growing segment of consumers. Therefore, there is a considerable interest in the development of preservative-free or self-preserving cosmetics. In these formulations traditional/chemical preservatives have been replaced by other cosmetic ingredients with antimicrobial properties that are not legislated as preservatives according to the Annex VI of the Commission Directive 76/768/EEC and the amending directives (2003/15/EC, 2007/17/EC and 2007/22/EC). 'Hurdle Technology', a technology that has been used for the control of product safety in the food industry since 1970s, has also been applied for the production of self preserving cosmetics. 'Hurdle Technology' is a term used to describe the intelligent combination of different preservation factors or hurdles to deteriorate the growth of microorganisms. Adherence to current good manufacturing practice, appropriate packaging, careful choice of the form of the emulsion, low water activity and low or high pH values are significant variables for the control of microbial growth in cosmetic formulations. This paper describes the application of the basic principles of 'Hurdle Technology' in the production of self-preserving cosmetics. Multifunctional antimicrobial ingredients and plant derived essential oils and extracts that are used as alternative or natural preservatives and are not listed in Annex VI of the Cosmetic Directive are also reported. PMID- 19302512 TI - Qualification of a precise and easy-to-handle sweat casting imprint method for the prediction and quantification of anti-perspirant efficacy. AB - A time- and cost-effective sweat casting method using the forearm as test site to assess the efficacy of several anti-perspirant formulations with a low number of test subjects has been evaluated and qualified. The imprint sweat casting method is based on a 2-component silcone-imprint technique to measure the efficacy of more than eight products in parallel with the same test subject. In studies using aluminum chlorohydrate (ACH) formulations as test anti-perspirants, a clear-cut correlation could be demonstrated between sweat gland activities measured by the imprint method and gravimetric measurement of sweat gland activities. Concentration-dependent inhibition of sweat gland activity could be observed with the imprint technique up to an ACH concentration of 15%, and all formulations containing 2% ACH or above resulted in statistically significant reduction of sweat gland activity (P < 0.001) when compared with untreated control areas. Furthermore, the SDs of individual studies using the imprint technique were in a range of +/-20% of sweat gland activity, which can be regarded rather low for in vivo measurements of a complex process like sweat secretion. A group-wise comparison between the measurements of anti-perspirant activity as determined by the imprint protocol and the Food and Drug Administration (FDA) Guideline compliant gravimetric hot-room protocol revealed that the test results for anti perspirant activity obtained with the imprint protocol are similar to those obtained with the hot-room protocol. Moreover, the data generated with the imprint protocol have a high predictive value for the outcome of a later guideline-compliant hot-room test. As the imprint casting method tends to be a little more sensitive for formulations with low anti-perspirant activity, and seems to be associated with less interassay variability than the standard gravimetric hot-room test, the imprint casting method may select products which later fail to pass the standard gravimetric hot-room test. Meanwhile the imprint sweat casting has proven to be a robust method useful to support efficacy oriented product development. Therefore, in later stages of utilization it might even evolve into an efficient claim substantiation tool. PMID- 19302513 TI - Suffering in silence: a qualitative meta-data-analysis of postpartum depression. AB - In this article, we apply a relational lens to a grounded theory meta-data analysis of qualitative studies on postpartum depression (PPD) conducted between 1999 and 2005. Women in all studies report feeling that they have failed to live up to cultural standards for a "good mother." Central to this experience is a sense that these negative feelings could not be spoken. The analysis shows how constructions of motherhood and the reactions of others combine with feelings of incompetence to precipitate isolation from others. Women survive depression through support that validates their experience and promotes eventual reconnection with others. Conclusions emphasize the need for persons trained to facilitate relational connection to develop interventions that address the interpersonal contexts of PPD. PMID- 19302514 TI - Implicit family process rules in eating-disordered and non-eating-disordered families. AB - Family environment has been shown to be one of the factors related to the presence of eating disorders among young-adult females. Clinical experience and theories about eating disorders postulate that implicit family rules are an intricate part of family process that may have a great effect on the creation and maintenance of such problems. This study compared implicit family process rules (specifically rules pertaining to kindness; expressiveness and connection; constraining thoughts, feelings, and self; inappropriate caretaking; and monitoring) in families with a young-adult female diagnosed with an eating disorder-either anorexia nervosa, bulimia nervosa, or eating disorder not otherwise specified-and families with a young-adult female without an eating disorder diagnosis. One hundred two families (51 eating disordered and 51 comparison) participated in the study. Mothers, fathers, young-adult female children, and siblings completed the Family Implicit Rules Profile (Harper, Stoll, & Larson, 2007). Results indicated that eating-disordered families are governed by a greater proportion of constraining family rules than are non-eating disordered families. Additionally, eating-disordered youth reported a lower proportion of facilitative family rules and a higher proportion of constraining family rules than did parents and siblings. Theoretical, research, and clinical implications are discussed. PMID- 19302515 TI - Psychological differences between HIV-positive mothers who disclose to all, some, or none of their biological children. AB - This study explored the psychological differences between HIV-positive women who disclosed their serostatus to all, some, or none of their biological children. Data from this project come from a larger, longitudinal investigation of the disclosure process of HIV-infected women. Data were obtained regarding the disclosure processes and the psychological adjustment of women with children (n = 90) based on measures of depression, anxiety, and stress. Results suggest that women who disclose to all, some, or none of their children are not significantly different with regard to emotional well-being. Implications for marriage and family therapists are provided. PMID- 19302516 TI - The Maudsley model of family-based treatment for anorexia nervosa: a qualitative evaluation of parent-to-parent consultation. AB - This article describes the qualitative analysis of a randomized control trial that explores the use of parent-to-parent consultations as an augmentation to the Maudsley model of family-based treatment for anorexia. Twenty families were randomized into two groups, 10 receiving standard treatment and 10 receiving an additional parent-to-parent consultation. Parents of all families were interviewed regarding their experience of treatment and transcripts were analyzed with the assistance of QSR N-Vivo. Parents described parent-to-parent consultations as an intense emotional experience that helped them to feel less alone, to feel empowered to progress, and to reflect on changes in family interactions. These results suggest that parent-to-parent consultations are seen as a useful augmentation to the Maudsley model of family-based treatment for anorexia nervosa. PMID- 19302517 TI - Clinical practice patterns of Canadian couple/marital/family therapists. AB - This clinical practice pattern survey had two unique aspects. It was a national survey of American Association for Marriage and Family Therapy (AAMFT) members in Canada that included all AAMFT membership categories, including student, affiliate, associate, clinical, and supervisor. It compared practice pattern data for clinical members from Canada and the United States. The results also showed that students, affiliates, and associates are very interested in identifying who they are as C/MFTs and that they are receiving various types of training. Clinical members in both Canada and the United States are fairly similar in terms of demographics and therapy practice, except when it comes to the models they identify as most influential. Future client satisfaction and outcome research is needed to determine how clients are specifically benefiting from therapy. PMID- 19302518 TI - Evaluating scholarship productivity in COAMFTE-accredited PhD programs. AB - Due to an increasing trend among states to cut higher education funds, many universities are relying more on private donations and federal funding to keep programs afloat. Scholarship productivity in general has become an integral factor in terms of universities granting tenure to faculty, allocating resources, and supporting program goals due to the fact that more research in a particular area tends to increase the likelihood that one will obtain funding from federal, state, and private sources. In the past, ranking systems have also been used to evaluate programs. However, most ranking systems use methodologies that do not quantify research productivity or evaluate factors that match current university trends. The purpose of this article is to explore current scholarship productivity trends among COAMFTE-accredited doctoral programs through the use of several evaluation methods. Specifically, productivity was examined in regard to the following areas: (a) family therapy journal publications; (b) family science journal publications; (c) historic journal publication trends; and (d) recent journal publication trends. PMID- 19302519 TI - A national survey of family physicians: perspectives on collaboration with marriage and family therapists. AB - Recognizing the fit between family medicine and marriage and family therapy (MFT), members of both fields have made significant advances in collaborative health research and practice. To add to this work, we surveyed a nationwide random sample of 240 family physicians (FPs) and asked about their perspectives and experiences of collaboration with MFTs. We found that FPs frequently perceive a need for their patients to receive MFT-related care, but their referral to and collaboration with MFTs were limited. Through responses to an open-ended question, we gained valuable information as to how MFTs could more effectively initiate collaboration with FPs. PMID- 19302520 TI - Resiliency in the practicing marriage and family therapist. AB - Although burnout in the helping professions is well documented, few studies have examined the phenomenon of the resilient therapist. This study used a grounded theory methodology to construct a theory of therapist resilience. The participants were eight licensed marital and family therapists: five females, three males, all Caucasian, with an average age of 58.9 and an average of 22.6 years of experience who reported feeling energized by the practice of therapy. The theory that was constructed included a central category (Integration of Self with Practice), a paradigm (Trust in Self), and two main categories (Career Development and Practice of Therapy). The process involved an initial calling, a positive agency experience, career corrections, the influence of relationships, and a move to a more flexible environment. PMID- 19302521 TI - Integrity of the marriage and family therapy research literature: perceptions and recommendations. AB - Reports of falsification, fabrication, plagiarism, and other violations of research integrity across the sciences are on the increase. Joining with other disciplines to actively protect the integrity of the marriage and family therapy (MFT) research literature is of utmost importance to both the discipline and the future of the profession. To inform the issues raised, results are presented of an informal survey among MFT clinical members on their perceptions about the literature together with their preferences for how best to protect its integrity. This article initiates an important discussion about the honesty of MFT research. PMID- 19302523 TI - Identification of novel putative virulence factors, adhesin AIDA and type VI secretion system, in atypical strains of fish pathogenic Edwardsiella tarda by genomic subtractive hybridization. AB - Edwardsiella tarda, which is known to be the causative agent of edwardsiellosis in freshwater and marine fish, has two motility phenotypes. Typical strains exhibiting motility are isolated mainly from freshwater fish and Japanese flounder. Atypical strains exhibiting non-motility are isolated mainly from marine fish, with the exception of Japanese flounder. Subtractive hybridization was performed to identify genomic differences between these two phenotypes. Two fragments which showed homology to potential virulence factors were isolated from atypical strains: the autotransporter adhesin AIDA and a component of T6SS. We analysed DNA sequences of about 5 kbp containing these fragments and identified two partial ORF, and ORF encoding for other components of T6SS. The predicted amino acid sequences showed remarkably low homology to components of T6SS reported in the typical E. tarda strain PPD130/91. Furthermore, the organization of these ORF was different from the gene cluster of the typical E. tarda strain. AIDA and T6SS may therefore be associated with different pathogenicity in typical and atypical E. tarda hosts. PMID- 19302524 TI - Analysis of cis-regulatory elements in the 5' untranslated region of murine leukemia virus controlling protein expression. AB - It has previously been reported by us that high-level expression of the Env protein of Fr-MLV clone A8 in brains is crucial for induction of spongiform neurodegeneration, and that the 0.3-kb fragment containing the R, U5, and the 5' leader sequence of A8 is responsible for neuropathogenicity. In the present study, the role of the 5' untranslated region in protein expression was investigated. Luciferase expression vectors containing the LTR (R-U3-U5) and 5' leader sequence of A8 and non-neuropathogenic 57 Fr-MLV, designated gl-A8 and gl 57, and their chimeric vectors, were constructed, and transfected into rat glial cells F10. Replacement of the region containing the 3' half of R, U5, and 5' leader sequence of gl-A8 with that of 57 showed a reduction in luciferase activities, and replacement of this region of gl-57 with that of A8 showed increased luciferase activity. These results show that the region containing the 3' half of R, U5, and 5' leader sequence of A8 more efficiently up-regulates protein expression than 57. In particular, the 3' half of 5' leader of A8 was most responsible for the up-regulation of protein expression. Of interest, after replacement of the fragments between A8 and 57, changes in the activities of vectors containing A8-U3 paralleled the amount of mRNA, but the activities of vectors containing 57-U3 did not. Furthermore, it is suggested that the region containing R, U5, and the 5' leader sequence influences transcriptional or post transcriptional steps, depending on the upstream sequence containing enhancer elements and promoter. PMID- 19302525 TI - Real-time polymerase chain reaction as a rapid and efficient alternative to estimation of picornavirus titers by tissue culture infectious dose 50% or plaque forming units. AB - Quantification of viral infectious units is traditionally measured by methods based on forming plaques in semisolid media (PFU) or endpoint dilution of a virus containing solution (TCID50), methods that are laborious, time-consuming and take on average 3-7 days to carry out. Quantitative real-time PCR is an established method to quantify nucleic acids at high accuracy and reproducibility, routinely used for virus detection and identification. In the present study, a procedure was developed using a two-step real-time PCR and the SYBR Green detection method to study whether there are correlations between TCID50/ml, PFU/ml and Ct values generated by real-time PCR enabling rapid and efficient calculation of titer equivalents when working with viruses in the research laboratory. In addition, an external standard with known concentrations was included using in vitro transcribed viral RNA, thus allowing the calculation of the amount of RNA copies needed for various applications (i.e. per plaque or TCID50).The results show that there is a correlation between the three quantification methods covering a wide range of concentration of viruses. Furthermore, a general regression line between TCID50 and Ct values was obtained for all viruses included in the study, which enabled recording titer equivalents using real-time PCR. Finally, by including an external standard, the amount of RNA genomes generating one TCID50 or PFU for each enterovirus serotype included was determined. PMID- 19302526 TI - Identification of multiple sites suitable for insertion of foreign genes in herpes simplex virus genomes. AB - Information on sites in HSV genomes at which foreign gene(s) can be inserted without disrupting viral genes or affecting properties of the parental virus are important for basic research on HSV and development of HSV-based vectors for human therapy. The intergenic region between HSV-1 UL3 and UL4 genes has been reported to satisfy the requirements for such an insertion site. The UL3 and UL4 genes are oriented toward the intergenic region and, therefore, insertion of a foreign gene(s) into the region between the UL3 and UL4 polyadenylation signals should not disrupt any viral genes or transcriptional units. HSV-1 and HSV-2 each have more than 10 additional regions structurally similar to the intergenic region between UL3 and UL4. In the studies reported here, it has been demonstrated that insertion of a reporter gene expression cassette into several of the HSV-1 and HSV-2 intergenic regions has no effect on viral growth in cell culture or virulence in mice, suggesting that these multiple intergenic regions may be suitable HSV sites for insertion of foreign genes. PMID- 19302527 TI - Engagement of Penicillium marneffei conidia with multiple pattern recognition receptors on human monocytes. AB - P. marneffei is a thermal dimorphic fungus which causes penicilliosis, an opportunistic infection in immunocompromised patients in South and Southeast Asia. Little is known about the innate immune response to P. marneffei infection. Therefore, the initial response of macrophages to P. marneffei conidia was evaluated by us. Adhesion between monocytes from healthy humans and fungal conidia was examined and found to be specifically inhibited by MAbs against PRR, such as MR, (TLR)1, TLR2, TLR4, TLR6, CD14, CD11a, CD11b, and CD18. To study the consequences of these interactions, cytokines were also examined by ELISA. Binding of P. marneffei conidia to monocytes was significantly inhibited, in a dose-dependent manner, by MAbs against MR, TLR1, TLR2, TLR4, TLR6, CD14, CD11b and CD18. When monocytes were co-cultured with the conidia, there was an increase in the amount of surface CD40 and CD86 expression, together with TNF-alpha and IL 1beta production, compared to unstimulated controls. In assays containing anti TLR4 or anti-CD14 antibody, reduction in the amount of TNF-alpha released by monocytes stimulated with P. marneffei conidia was detected. In addition, it was found that production of TNF-alpha and IL-1beta from adherent peripheral blood monocytes was partially impaired when heat-inactivated autologous serum, in place of untreated autologous serum, was added to the assay. These results demonstrate that various PRR on human monocytes participate in the initial recognition of P. marneffei conidia, and the engagement of PRR could partly initiate proinflammatory cytokine production. PMID- 19302528 TI - Immunolocalization and serum antibody responses to Brugia malayi pepsin inhibitor homolog (Bm-33). AB - cDNA coding for Brugia malayi pepsin inhibitor homolog (Bm-33) from the human filarial parasite was cloned in pRSET for large-scale expression and functional characterization. The pRSET-B cloned gene did not yield recombinant protein expression and the reason was attributed to the presence of an N-terminal signal peptide. The gene was subcloned in pRSET-A without a signal peptide and the 33 kDa histidine-tagged recombinant protein was purified by IMAC. All individuals from an endemic area generated IgG responses against Bm-33 in the order MF>CP>EN. Isotype analysis indicated an elevated IgG4 reactivity in the order MF>EN>CP. Bm 33-specific IgE levels were elevated in MF, CP and EN compared to non-endemic normals with no significant differences among the groups. Paraffin-embedded sections of Setaria digitata (cattle filarial parasite) stained with mouse anti Bm-33 antibodies exhibited the hypodermal nature of Bm-33. These findings suggest that Bm-33 is an immunodominant antigen and contributes to filarial pathogenesis. PMID- 19302529 TI - Phenotypic characteristics and pathogenic ability across distinct morphotypes of Burkholderia pseudomallei DT. AB - Burkholderia pseudomallei DT is unusual as it exhibits six distinct colony morphotypes. Types III and V show stronger motility, whereas type VI exhibits the highest levels of bacterial association with peritoneal exudate cells. Although the bacterial loads in the organs are not significantly different for infections by the six distinct morphotypes, higher mortality (100% and 89%, respectively) and larger areas of abnormal liver debris (20.6% and 22.4%, respectively) are found with types I- and III-infected mice compared to the others. These morphotypes sometimes undergo switching to a mucoid type in the body of mice, but the reverse has never been observed. PMID- 19302530 TI - Expression of c-MET, low-molecular-weight cytokeratin, matrix metalloproteinases 1 and -2 in spinal chordoma. AB - AIMS: In skull base chordoma, c-MET expression has been reported to correlate with younger patient age and favourable prognosis; however, it also contributes to tumour invasiveness, especially in recurrent lesions, suggesting variable roles for c-MET according to clinical status. The aim of this study was to investigate the significance of c-MET expression in spinal chordoma, which affects patients who are 10-20 years older than those with skull base chordoma. METHODS AND RESULTS: Using immunohistochemical techniques, the expression of c MET and its ligand, hepatocyte growth factor (HGF) was investigated in 34 primary spinal chordomas and compared with other clinicopathological parameters. Expression of c-MET and HGF was observed in 85.3 and 21.7% of lesions, respectively. c-MET expression correlated with the expression of an epithelial marker, low-molecular-weight cytokeratin (CAM5.2). Lesions with higher c-MET expression showed significantly stronger expression of proteinases, including matrix metalloproteinase (MMP)-1 and MMP-2. However, c-MET expression was not associated with patient age, proliferative ability estimated by MIB-1 labelling index, or prognosis. CONCLUSIONS: c-MET expression was observed in most spinal chordomas and correlated with the expression of CAM5.2, suggesting a relationship to an epithelial phenotype. PMID- 19302531 TI - A persistent scalp nodule caused by the human botfly: exoskeletal larval spines are a key diagnostic feature of myiasis. PMID- 19302532 TI - Re: Adenocarcinoma arising in a background of chronic atrophic pouchitis in an ileoanal pouch for ulcerative colitis. PMID- 19302533 TI - Immunohistochemical stain for cytokeratin 7, S100A1 and claudin 8 is valuable in differential diagnosis of chromophobe renal cell carcinoma from renal oncocytoma. PMID- 19302534 TI - Expression and molecular analysis of c-kit and PDGFRs in Kaposi's sarcoma of different stages and epidemiological settings. PMID- 19302535 TI - Re: Interobserver agreement in grading of colorectal cancer--findings from a nationwide web-based survey of histopathologists. PMID- 19302536 TI - Sacrococcygeal teratoma with extensive nephrogenic differentiation: a rare finding not to be misdiagnosed as yolk sac tumour. PMID- 19302537 TI - Interstitial pneumonitis caused by Pneumocystis jirovecii pneumonia (PCP) during bortezomib treatment. PMID- 19302538 TI - Emerin immunohistochemistry reveals diagnostic features of nuclear membrane arrangement in thyroid lesions. AB - AIMS: Objective appreciation of irregularities of the nuclear shape is a key parameter in the diagnosis of thyroid lesions, since foldings of the nuclear membrane (NM) featuring indentations, grooves and pseudoinclusions characterize papillary thyroid carcinomas (PTC). The aim was to focus on the immunohistochemistry of emerin, a protein of the NM whose decoration best demarcates the nuclear shape. METHODS AND RESULTS: Immunohistochemistry of the NM with emerin as well as three-dimensional reconstruction of the images (through deconvolution processing) performed on a series of 54 cases (processed following the tissue array procedure) revealed a uniform arrangement of the NM in non neoplastic lesions (thyroiditis, microfollicular goitre, follicular adenoma) and normal thyroid as well as in follicular carcinoma. In contrast, irregular folding of the membrane and presence of curling and invaginations, eventually leading to the formation of nuclear pseudoinclusions, was observed in PTC cells. CONCLUSIONS: Decoration of the NM represents an original approach to identify PTC nuclear shape, highlights new structural features and might be helpful in the differential diagnosis between so-called nuclear pseudoinclusions and artefactual 'bubbles'. PMID- 19302539 TI - Architecture of developing multicellular yeast colony: spatio-temporal expression of Ato1p ammonium exporter. AB - Yeasts, when growing on solid surfaces, form organized multicellular structures, colonies, in which cells differentiate and thus possess different functions and undergo dissimilar fate. Understanding the principles involved in the formation of these structures requires new approaches that allow the study of individual cells directly in situ without needing to remove them from the microbial community. Here we introduced a new approach to the analysis of whole yeast microcolonies either containing specific proteins labelled by fluorescent proteins or stained with specific dyes, by two-photon excitation confocal microscopy. It revealed that the colonies are covered with a thin protective skin like surface cell layer which blocks penetration of harmful compounds. The cells forming the layer are tightly connected via cell walls, the presence of which is essential for keeping of protective layer function. Viewing the colonies from different angles allowed us to reconstruct a three-dimensional profile of the cells producing ammonium exporter Ato1p within developing microcolonies growing either as individuals or within a group of microcolonies. We show that neighbouring microcolonies coordinate production of Ato1p-GFP. Ato1p itself appears synchronously in cells, which do not originate from the same ancestor, but occupy specific position within the colony. PMID- 19302540 TI - Effect of gelling agent on colony formation in solid cultivation of microbial community in lake sediment. AB - Since Robert Koch and colleagues found agar to be an effective gelling agent over a century ago, the pure culture method using agar plates has long been a standard of microbiology. Agar is undoubtedly easy to handle and useful for culture of microorganisms, but recent discovery of the ubiquity of microorganisms that cannot be cultured on agar raises a question: is agar really the best agent? In this study, we investigated the effect of two gelling agents, agar and gellan gum, on colony formation of a diverse array of microorganisms (total 108 strains) newly isolated from freshwater sediments and a representative microorganism as a slow grower on agar medium, Gemmatimonas aurantiaca, to clarify (i) whether they can grow on both agar and gellan gum plates, and (ii) the difference in time required for colony formation between the two gelling agents. Interestingly, 22 of 108 isolates showed no ability to form any visible colonies on the agar medium but did so on the gellan gum medium, and showed low 16S rRNA gene sequence similarities to their closest species. The remaining 86 isolates grew on both agar and gellan gum, but 52 of them grew much faster on gellan gum than on agar. Moreover, gellan gum also significantly stimulated the colony formation of the representative slow-growing microorganism G. aurantiaca. Our results demonstrate that the gelling agent is a crucial factor for the growth of bacteria on plate media, and that alternatives to agar will be very important for increasing the culturability of yet-to-be cultured microorganisms. PMID- 19302541 TI - Metagenomic signatures of 86 microbial and viral metagenomes. AB - Previous studies have shown that dinucleotide abundances capture the majority of variation in genome signatures and are useful for quantifying lateral gene transfer and building molecular phylogenies. Metagenomes contain a mixture of individual genomes, and might be expected to lack compositional signatures. In many metagenomic data sets the majority of sequences have no significant similarities to known sequences and are effectively excluded from subsequent analyses. To circumvent this limitation, di-, tri- and tetranucleotide abundances of 86 microbial and viral metagenomes consisting of short pyrosequencing reads were analysed to provide a method which includes all sequences that can be used in combination with other analysis to increase our knowledge about microbial and viral communities. Both principal component analysis and hierarchical clustering showed definitive groupings of metagenomes drawn from similar environments. Together these analyses showed that dinucleotide composition, as opposed to tri- and tetranucleotides, defines a metagenomic signature which can explain up to 80% of the variance between biomes, which is comparable to that obtained by functional genomics. Metagenomes with anomalous content were also identified using dinucleotide abundances. Subsequent analyses determined that these metagenomes were contaminated with exogenous DNA, suggesting that this approach is a useful metric for quality control. The predictive strength of the dinucleotide composition also opens the possibility of assigning ecological classifications to unknown fragments. Environmental selection may be responsible for this dinucleotide signature through direct selection of specific compositional signals; however, simulations suggest that the environment may select indirectly by promoting the increased abundance of a few dominant taxa. PMID- 19302542 TI - Looking "back to the future". PMID- 19302543 TI - Nurses' uncertainty in decision-making: a literature review. AB - AIM: This paper is a report of the results of a review of the literature conducted with the goal of determining how nurses' clinical uncertainty has been conceptualized in the nursing literature. BACKGROUND: Although existing research has advanced the body of knowledge regarding the concept of uncertainty in decision-making, this has been largely from physicians' viewpoints and from patients' perspectives (patients' uncertainty). Understanding how nurses' experience and act on uncertainty remains relatively unreported. METHOD: A search of Medline, CINAHL, and PubMed databases was conducted to retrieve literature published from 1990 to 2007. The question guiding the literature review was: How has nurses' clinical uncertainty been conceptualized in nursing literature? FINDINGS: Little exploration has been done of nurses' experience of uncertainty in practice. Many investigators have not theorized about the uncertainty in their studies, but have described nurses' uncertainty in the context of clinical decision-making. The findings from these studies indicated that unfamiliarity with the aspects of patient care is a source of uncertainty, and nurses tended to rely on heuristics or on the expertise of colleagues as sources of information for practice decisions. Expressing uncertainties as information needs might help guide information seeking and reduce uncertainty. However, studies indicated that nurses have difficulty recognizing or expressing uncertainties, and as a result, information needs are not recognized and information seeking is not initiated. CONCLUSIONS: A more comprehensive understanding of nurses' uncertainty could lead to the development and implementation of strategies to support nurses in their clinical decision-making and practice. Descriptions are needed about how nurses experience and respond to uncertainty in their practice, and the influence of uncertainty on their information needs and information seeking. PMID- 19302544 TI - Use of evidence-based practice in school nursing: prevalence, associated variables, and perceived needs. AB - BACKGROUND: Research on the adoption of evidence-based practice (EBP) in health care has been focused mainly on hospital settings and primary care; little is known about EBP adoption and implementation among school nurses in the United States (US). OBJECTIVES: The objectives of this study were to (1) describe the prevalence of EBP use among school nurses, (2) describe demographic, individual, and organizational factors associated with EBP use, and (3) identify resources needed to enhance EBP use. METHODS: A survey designed for this study was mailed to all school nurses in Iowa in the US, with a response rate of 56.8% (n= 386). Descriptive statistics and t-tests were used to indicate and compare the variables of interest. Multiple regression and analysis of variance (ANOVA) methods were used to identify variables explaining the variance in the use of EBP. RESULTS: Information sources, professional membership, and district size explain 22% of the variance in current use of EBP. Identified resources needed to increase the use of EBP included networking opportunities, predeveloped EBP guidelines, and education on outcome evaluation. CONCLUSIONS: This study indicates important information on variables that affect current practice in school nursing and also shows requested resources for increasing EBP use. These results can be used to develop translation strategies to increase the use of EBP in the school setting. Because only 22% of the variance was explained, further research is needed to identify additional variables. PMID- 19302545 TI - Comparisons of acute physiological parameters influencing outcome in patients with traumatic brain injury and hemorrhagic stroke. AB - BACKGROUND: Although several factors are known to influence prognosis after acute brain injury, physiologic derangements were found to be associated with, or contributing to, increased mortality and disability in these patients. AIMS: This study was conducted to identify acute physiological parameters that significantly predict outcome and compare the identified parameters in patients with traumatic brain injury (TBI) and hemorrhagic stroke. METHODS: Participants were 190 adult patients with TBI or hemorrhagic stroke who were admitted within 24 hours of injury. Details concerning potential physiological predictors were collected, that is, blood pressure (BP), heart rate, respiratory rate, body temperature, hematocrit, partial pressure of oxygen and carbon dioxide in arterial blood, serum osmolality, pH, total cholesterol, and glucose levels on admission. Mortality and functional and cognitive ability 6 months after injury were measured as outcome variables. RESULTS: Most physiological parameters, with the exceptions of blood glucose, total cholesterol, and osmolality, were significant prognostic predictors in TBI and hemorrhagic stroke. However, the physiological parameters and their predictabilities differed in patients with TBI and hemorrhagic stroke, which indicated that physiological predictors depend on chosen outcome and brain injury type. CONCLUSIONS AND IMPLICATIONS: In critical care units, the most important aims of nursing care are decreasing mortality and disability rates. In order to achieve such goals, acute physiological parameters such as BP, respiratory rate, heart rate, PaO2, PaCO2, and temperature should be monitored and managed, with the aim of reducing the risk of secondary brain injury. PMID- 19302548 TI - Diversities of coral-associated bacteria differ with location, but not species, for three acroporid corals on the Great Barrier Reef. AB - Patterns in the diversity of bacterial communities associated with three species of Acropora (Acropora millepora, Acropora tenuis and Acropora valida) were compared at two locations (Magnetic Island and Orpheus Island) on the Great Barrier Reef to better understand the nature and specificity of coral-microbial symbioses. Three culture-independent techniques demonstrated consistent bacterial communities among replicate samples of each coral species, confirming that corals associate with specific microbiota. Profiles were also conserved among all three species of Acropora within each location, suggesting that closely related corals of the same genus harbor similar bacterial types. Bacterial community profiles of A. millepora at Orpheus Island were consistent in samples collected throughout the year, indicating a stable community despite temporal changes. However, DGGE and T-RFLP profiles differed on corals from different reefs. Nonmetric multidimensional scaling of T-RFLP profiles showed that samples grouped according to location rather than coral species. Although similar sequences were retrieved from clone libraries of corals at both Magnetic and Orpheus Island, differences in the relative dominant bacterial ribotypes within the libraries drive bacterial community structure at different geographical locations. These results indicate certain bacterial groups associated specifically with corals, but the dominant bacterial genera differ between geographically-spaced corals. PMID- 19302549 TI - Combined monitoring of changes in delta13CH4 and archaeal community structure during mesophilic methanization of municipal solid waste. AB - Reconstituted municipal solid waste (MSW) with varying contents of putrescible and cellulosic waste was incubated anaerobically under mesophilic conditions. Standard physicochemical parameters were monitored, together with stable isotopic signatures of produced CH(4) and CO(2). delta(13)C values for CH(4) indicated a change of methanogenic metabolism with time. CH(4) was predominantly produced from H(2)/CO(2) at the beginning of the incubations. This period was associated with important shifts in archaeal communities monitored by automated ribosomal intergenic spacer analysis (ARISA) and FISH of oligonucleotidic probes targeting specifically 16S rRNA gene of various methanogenic groups. The onset of the active methane generation phase was characterized by an increase of CH(4)delta(13)C, indicating a progressive shift toward an aceticlastic metabolism. When the methane production levelled off, a decrease in the isotopic signature was observed toward values characteristics of hydrogenotrophic metabolism. ARISA profiles were, however, found to be stable from the beginning of the active methane generation phase until the end of the experiment. FISH observation indicated that members of the family Methanosarcinaceae were predominant in the archaeal community during this period, suggesting that these methanogens might exhibit a high metabolic versatility during methanization of waste. PMID- 19302550 TI - Comparative assessment of human and farm animal faecal microbiota using real-time quantitative PCR. AB - Pollution of the environment by human and animal faecal pollution affects the safety of shellfish, drinking water and recreational beaches. To pinpoint the origin of contaminations, it is essential to define the differences between human microbiota and that of farm animals. A strategy based on real-time quantitative PCR (qPCR) assays was therefore developed and applied to compare the composition of intestinal microbiota of these two groups. Primers were designed to quantify the 16S rRNA gene from dominant and subdominant bacterial groups. TaqMan probes were defined for the qPCR technique used for dominant microbiota. Human faecal microbiota was compared with that of farm animals using faecal samples collected from rabbits, goats, horses, pigs, sheep and cows. Three dominant bacterial groups (Bacteroides/Prevotella, Clostridium coccoides and Bifidobacterium) of the human microbiota showed differential population levels in animal species. The Clostridium leptum group showed the lowest differences among human and farm animal species. Human subdominant bacterial groups were highly variable in animal species. Partial least squares regression indicated that the human microbiota could be distinguished from all farm animals studied. This culture-independent comparative assessment of the faecal microbiota between humans and farm animals will prove useful in identifying biomarkers of human and animal faecal contaminations that can be applied to microbial source tracking methods. PMID- 19302552 TI - Effect of inhibitors of mitogen-activated protein kinase kinase on alpha(1B) adrenoceptor phosphorylation. AB - 1 Mitogen-activated protein kinases mediate hormone/neurotransmitter action on proliferation and differentiation and participate in receptor regulation. The effect of inhibitors of mitogen-activated kinase kinase (MEK) on alpha(1B) adrenoceptor phosphorylation state and function was studied using different cell lines. It was observed that at nanomolar concentrations the MEK inhibitors, PD98059 (2'-amino-3'-methoxyflavone) and UO126 [1,4-(diamino-2,3-dicyano/1,4-bis (2-aminophenylthio)-butadiene], increased alpha(1B)-adrenoceptor phosphorylation and diminished the functional response of this receptor to noradrenaline. These agents did not alter the action of lysophosphatidic acid. 2 Staurosporine (IC(50) approximately 0.8 nm) (a general protein kinase inhibitor) and bis-indolyl maleimide I (IC(50) approximately 200 nm) (a selective protein kinase C inhibitor) inhibited PD98059-induced alpha(1B)-adrenoceptor phosphorylation. In contrast, neither wortmannin (phosphoinositide 3-kinase inhibitor) nor genistein (protein tyrosine kinase inhibitor) had any effect. The data suggest the possibility that MEK might exert control on the activity of the enzymes that regulate receptor phosphorylation, such as G-protein-coupled receptor kinases, protein kinase C or serine/threonine protein phosphatases. 3 Coimmunoprecipitation studies showed a constant association of total extracellular signal-regulated kinase 2 (ERK2) with alpha(1B)-adrenoceptors. Association of phospho-ERK 1/2 to alpha(1B)-adrenoceptors increased not only in response to agonist but also in response to agents that increase alpha(1B) adrenoceptor and ERK1/2 phosphorylation [such as endothelin-1, phorbol 12 myristate-13-acetate (PMA) and epidermal growth factor (EGF)]; not surprisingly, PD98059 decreased this effect. 4 Our data show that blockade of MEK activity results in increased alpha(1B)-adrenoceptor phosphorylation, diminished adrenoceptor function and perturbation of receptor-ERK1/2 interaction. PMID- 19302551 TI - Role of 20-hydroxyeicosatetraenoic acid in altering vascular reactivity in diabetes. AB - 1 This study examined the role of 20-hydroxyeicosatetraenoic (20-HETE) in altering vascular function in streptozotocin (STZ)-induced diabetic rats. 2 The expression of CYP4A protein and the formation of 20-HETE were elevated in the kidney, but not in the renal or mesenteric vasculature, of diabetic animals. The vasoconstrictor responses to norepinephrine (NE), endothelin-1 (ET-1), and angiotensin II (Ang II) were significantly enhanced in the isolated perfused mesenteric vascular bed and renal artery segments of diabetic rats. Chronic treatment of the diabetic rats with 1-aminobenzotriazole (ABT, 50 mg kg(-1) alt( 1) diem) or N-hydroxy-N'-(4-butyl-2-methylphenyl) formamidine (HET0016, 2.5 mg kg(-1) day(-1)) attenuated the responses to these vasoconstrictors in both vascular beds. 3 The synthesis of 20-HETE in renal microsomes was reduced by >80% confirming that the doses of ABT and HET0016 were sufficient to achieve system blockade. Addition of HET0016 (1 microM) in vitro also normalized the enhanced vascular responsiveness of renal and mesenteric vessels obtained from diabetic animals to NE and inhibited the formation of 20-HETE by >90% while having no effect on the formation of epoxides. Vasodilator responses to carbachol and histamine were reduced in the mesenteric vasculature, but not in renal arteries, of diabetic rats. Treatment of the diabetic animals with HET0016 improved vasodilator responses in both vascular beds. Vascular sensitivity to exogenous 20 HETE was elevated in the mesenteric bed of diabetic animals compared to controls. 4 These results suggest that 20-HETE contributes to the elevation in vascular reactivity in diabetic animals. This effect is not due to increased vascular expression of CYP4A but may be related to either enhanced agonist-induced release of substrate (arachidonic acid) by the CaMKII/Ras-GTPase system and/or elevated vascular responsiveness to 20-HETE by the CaMKII/Ras-GTPase system and/or elevated vascular responsiveness to 20-HETE. PMID- 19302553 TI - Influence of high dietary sodium intake on the functional subtypes of alpha adrenoceptors in the renal cortical vasculature of Wistar-Kyoto rats. AB - 1 Increased renal vascular resistance is one renal functional abnormality that contributes to hypertension, and alpha(1)-adrenoceptors play a pivotal role in modulating this renal vascular resistance. This study investigates the functional contribution of alpha(1)-adrenoceptor subtypes in the renal cortical vasculature of Wistar-Kyoto rats on a normal sodium diet (WKYNNa) compared with those given saline to drink for 6 weeks (WKYHNa). 2 The renal cortical vascular responses to the adrenergic agonists noradrenaline (NA), methoxamine (ME) and phenylephrine (PE) were measured in WKYHNa and WKYNNa rats either in the absence (the control phase) or presence of chloroethylclonidine (CEC), an alpha(1B)-adrenoceptor antagonist, 5-methylurapidil (5-MeU), an alpha(1A) antagonist, or BMY7378, an alpha(1D) antagonist. 3 Results showed a greater renal cortical vascular sensitivity to NA, PE and ME in the WKYHNa compared with WKYNNa rats (P < 0.05). Moreover, 5-MeU and BMY7378 attenuated adrenergically induced renal cortical vasoconstriction in WKYHNa and WKYNNa rats; this response was largely blunted in CEC-treated WKYHNa rats (all P < 0.05) but not in CEC-treated WKYNNa rats. 4 The data suggest that irrespective of dietary sodium content, in Wistar-Kyoto rats alpha(1A)- and alpha(1D)-subtypes are the major alpha(1)-adrenoceptors in renal cortical vasculature; however, there appears to be a functional involvement of alpha(1B)-adrenoceptors in the WKYHNa rats. PMID- 19302554 TI - Cytochrome P450 metabolites of arachidonic acid play a role in the enhanced cardiac dysfunction in diabetic rats following ischaemic reperfusion injury. AB - 1 This study examined the contribution of cytochrome P450 metabolites of arachidonic acid in mediating ischaemia/reperfusion (I/R)-induced cardiac dysfunction in normal and diabetic rats. 2 We first compared the metabolism of arachidonic acid in microsomes prepared from the hearts of control rats and rats treated with streptozotocin (55 mg kg(-1)) to induce diabetes. The production of dihydroxyeicosatrienoic acids and epoxyeicosatrienoic acids (EETs) were similar in microsomes prepared from the hearts of control and diabetic rats, but the production of 20-hydroxyeicosatetraenoic acid (20-HETE) was two-fold higher in diabetic hearts than in control animals. 3 We then compared the change in left ventricular pressure (P(max)), left ventricular end-diastolic pressure, coronary flow and coronary vascular resistance in isolated perfused hearts obtained from control and diabetic animals after 40 min of global ischaemia (I) followed by 30 min of reperfusion (R). The decline in cardiac function was three- to five-fold greater in the hearts obtained from diabetic vs. control animals. 4 Pretreatment of the hearts with N-hydroxy-N'-(4-butyl-2-methyl-phenyl)-formamidine (HET0016, 1 microm), a selective inhibitor of the synthesis of 20-HETE, for 30 min before I/R resulted in significant improvement in the recovery of cardiac function in the hearts obtained from diabetic but not in control rats. Perfusion with an inhibitor of soluble epoxide hydrolase, 1-cyclohexyl-3-dodecyl urea (CDU), before I/R improved the recovery of cardiac function in hearts obtained from both control and diabetic animals. Perfusion with both HET0016 and CDU resulted in significantly better recovery of cardiac function of diabetic hearts following I/R than that seen using either drug alone. Pretreatment of the hearts with glibenclamide (1 microm), an inhibitor of ATP-sensitive potassium channels, attenuated the cardioprotective effects of both CDU and HET0016. 5 This is the first study to suggest that acute blockade of the formation of 20-HETE and/or reduced inactivation of EETs could be an important strategy to reduce cardiac dysfunction following I/R events in diabetes. PMID- 19302555 TI - Effect of central serotonin depletion on 5-HT receptor-mediated vasomotor responses in the middle meningeal artery of anaesthetized rats. AB - 1 It has been hypothesized that craniovascular 5-HT receptors mediating dilatation of cranial vessels undergo sensitization on decreased serotonergic transmission in migraine. This study analysed the effect of chemical lesion of the 5-HT system in the brain with 5,7-dihydroxytryptamine (5,7-DHT) on 5-HT receptor-mediated dilator responses to 5-carboxamidotryptamine (5-CT) in the middle meningeal artery of anaesthetized rats. 5-CT has recently been shown to elicit dilator responses in this cranial vessel via 5-HT(7) receptors and, to a much lesser extent, 5-HT(1B/1D) receptors. 2 Pretreatment with 5,7-DHT produced a drastic and selective decrease of 5-HT levels in the brain (78 +/- 6% and 94 +/- 2% in dorsal raphe and hypothalamic paraventricular nuclei, respectively) compared with controls (1% ascorbic acid). 3 Topical application of 5-CT (1-1000 microm) to exposed dura mater encephali produced concentration-dependent decreases in diastolic blood pressure and dilator responses in the middle meningeal artery that were similar in vehicle- and 5,7-DHT-pretreaed animals. 4 Hypotensive and meningeal dilator responses to 5-CT were unaltered by the 5 HT(1B/1D) receptor antagonist, GR-127935 (1 mg kg(-1), i.v.), but were strongly inhibited by the 5-HT(7) receptor antagonist, SB-269970 (1 mg kg(-1), i.v.), with similar efficacy, in both groups of animals. Treatment with GR-127935 + SB-269970 (1 mg kg(-1), i.v. each), produced a stronger inhibitory effect than individual treatments on hypotensive but not on meningeal responses to 5-CT. Meningeal 5 HT(7) receptor-mediated responses (i.e. in GR-127935-pretreated animals) were unchanged by 5,7-DHT pretreatment. 5 Results suggest that the sensitivity of craniovascular 5-HT(7) receptors mediating dilatation is unaffected by a decrease of 5-HT levels in the brain. A neuronal involvement of 5-HT in migraine seems more likely, therefore. PMID- 19302556 TI - Serum vascular endothelial growth factor in adult haematological patients with neutropenic fever: a comparison with C-reactive protein. AB - OBJECTIVES: Vascular endothelial growth factor (VEGF) is considered to be of importance in patients with sepsis. No data are available on VEGF kinetics in haematological patients with neutropenic fever. METHODS: Forty-two haematological patients were included into this prospective study. Median age was 57 yr (range 18-70). Fifteen patients received therapy for acute myeloid leukaemia and 27 patients received autologous stem cell transplantation for haematological malignancy. Laboratory samples for the determination of C-reactive protein (CRP) and VEGF were collected at the start of fever (d0) and then daily. RESULTS: The median serum VEGF concentrations were low in all study patients. In patients with severe sepsis (n = 5) the median VEGF on d0 was higher than in septic patients without signs of hypoperfusion or hypotension (n = 37) (77 pg/mL vs. 52 pg/mL, P = 0.061). Also on d1 the median VEGF concentration was higher in patients with severe sepsis (82 pg/mL vs. 56 pg/mL, P = 0.048). There were no statistically significant differences in CRP values on any day during the study period between patients with severe sepsis and those without. Time from d0 to the peak VEGF concentration (mean 1.02, SE 0.18 d) was shorter than that to the peak CRP concentration (mean 1.93, SE 0.15 d) (P = 0.002). CONCLUSION: Compared to CRP, serum VEGF was a more rapid indicator for sepsis in our haematological patients with neutropenic fever. Those with severe sepsis had higher VEGF concentrations than those without on d0 and d1 after the onset of fever. Further studies on VEGF are warranted in haematological patients. PMID- 19302557 TI - Risk factors for severe infection in patients with hairy cell leukemia: a long term study of 73 patients. AB - Although the survival of patients with hairy cell leukemia (HCL) has been improved by the therapeutic introduction of interferon alpha and purine analogs, it is still worsened by complications such as severe infections. In this long term study, we identified factors influencing patient outcomes in 73 patients with HCL. Median age at diagnosis was 53 yr and the gender ratio (M/F) was 2.3. At the time of HCL diagnosis, 60 patients (82%) were symptomatic and 22 of these had an infection. After a median follow-up of 13 yr, eight patients had died of secondary cancer (n = 2), HCL progression (n = 1) and age-related complications (n = 5). The 10-yr overall survival (OS), progression-free survival and relapse rates were 91 +/- 3%, 14 +/- 5% and 87 +/- 5%, respectively. In multivariate analyses, age >53 yr was the only factor adversely influencing OS and secondary cancer incidence, with adjusted hazard ratio (HR) of 9.30 (95%CI, 1.15-76.6; P = 0.037) and 2.80 (95%CI, 1.05-7.71; P = 0.04), respectively. Eleven patients developed severe infections. Absolute lymphocyte count (<1 x 10(9)/L) at diagnosis was the only factor influencing the occurrence of severe infections, with an adjusted HR of 4.01 (P = 0.007). Strikingly, we did not observe any significant correlation between neutrophil or monocyte counts and the incidence of infection. We confirmed long-term survival in HCL but found a high incidence of infection - even late in the course of the disease. The absolute lymphocyte count at diagnosis is a risk factor for the occurrence of severe infections. In addition to careful monitoring of infections, prompt initiation of anti-HCL treatment should be considered in patients with low lymphocyte counts. PMID- 19302558 TI - Danish CLL2-Study revisited: FISH on a cohort with a 20-yr follow-up confirms the validity of the hierarchical model of genomic aberrations in chronic lymphocytic leukaemia. PMID- 19302559 TI - Lenalidomide in combination with dexamethasone at first relapse in comparison with its use as later salvage therapy in relapsed or refractory multiple myeloma. AB - This subset analysis of data from two phase III studies in patients with relapsed or refractory multiple myeloma (MM) evaluated the benefit of initiating lenalidomide plus dexamethasone at first relapse. Multivariate analysis showed that fewer prior therapies, along with beta(2)-microglobulin (< or = 2.5 mg/L), predicted a better time to progression (TTP; study end-point) with lenalidomide plus dexamethasone treatment. Patients with one prior therapy showed a significant improvement in benefit after first relapse compared with those who received two or more therapies. Patients with one prior therapy had significantly prolonged median TTP (17.1 vs. 10.6 months; P = 0.026) and progression-free survival (14.1 vs. 9.5 months, P = 0.047) compared with patients treated in later lines. Overall response rates were higher (66.9% vs. 56.8%, P = 0.06), and the complete response plus very good partial response rate was significantly higher in first relapse (39.8% vs. 27.7%, P = 0.025). Importantly, overall survival was significantly prolonged for patients treated with lenalidomide plus dexamethasone with one prior therapy, compared with patients treated later in salvage (median of 42.0 vs. 35.8 months, P = 0.041), with no differences in toxicity, dose reductions, or discontinuations despite longer treatment. Therefore, lenalidomide plus dexamethasone is both effective and tolerable for second-line MM therapy and the data suggest that the greatest benefit occurs with earlier use. PMID- 19302560 TI - Ion-binding properties of Calnuc, Ca2+ versus Mg2+--Calnuc adopts additional and unusual Ca2+-binding sites upon interaction with G-protein. AB - Calnuc is a novel, highly modular, EF-hand containing, Ca(2+)-binding, Golgi resident protein whose functions are not clear. Using amino acid sequences, we demonstrate that Calnuc is a highly conserved protein among various organisms, from Ciona intestinalis to humans. Maximum homology among all sequences is found in the region that binds to G-proteins. In humans, it is known to be expressed in a variety of tissues, and it interacts with several important protein partners. Among other proteins, Calnuc is known to interact with heterotrimeric G-proteins, specifically with the alpha-subunit. Herein, we report the structural implications of Ca(2+) and Mg(2+) binding, and illustrate that Calnuc functions as a downstream effector for G-protein alpha-subunit. Our results show that Ca(2+) binds with an affinity of 7 mum and causes structural changes. Although Mg(2+) binds to Calnuc with very weak affinity, the structural changes that it causes are further enhanced by Ca(2+) binding. Furthermore, isothermal titration calorimetry results show that Calnuc and the G-protein bind with an affinity of 13 nm. We also predict a probable function for Calnuc, that of maintaining Ca(2+) homeostasis in the cell. Using Stains-all and terbium as Ca(2+) mimic probes, we demonstrate that the Ca(2+)-binding ability of Calnuc is governed by the activity based conformational state of the G-protein. We propose that Calnuc adopts structural sites similar to the ones seen in proteins such as annexins, c2 domains or chromogrannin A, and therefore binds more calcium ions upon binding to Gialpha. With the number of organelle-targeted G-protein-coupled receptors increasing, intracellular communication mediated by G-proteins could become a new paradigm. In this regard, we propose that Calnuc could be involved in the downstream signaling of G-proteins. PMID- 19302561 TI - Molecular composition and biological activity of commercial birch pollen allergen extracts. AB - BACKGROUND: Commercial extracts used for diagnosis and treatment of allergy are currently prepared from natural allergen sources. The aim of this study was to analyse birch pollen allergen extracts produced for in vivo diagnosis of birch pollen allergy regarding their contents of individual birch pollen allergens (Bet v 1, Bet v 2 and Bet v 4). METHODS: Protein contents were measured and the allergen composition was analysed by immunoblotting using antibody probes specific for Bet v 1, Bet v 2 and Bet v 4 in birch pollen extracts from five manufacturers of allergen extracts. The contents of the major birch pollen allergen, Bet v 1, were quantified with a specific two-site binding enzyme-linked immunosorbent assay with nanogram sensitivity for Bet v 1. The biological activities of the allergen extracts were evaluated by skin prick testing in birch pollen allergic patients and compared with their sensitization profiles. RESULTS: A more than 10-fold variation regarding total protein contents (23.1-314 microg mL(-1)) and also regarding the amounts of the major birch pollen allergen, Bet v 1 (1.62-19.6 microg mL(-1)) was found. The highly cross-reactive Bet v 4 allergen was absent in three of the five tested extracts. Furthermore, varying skin test results were obtained in birch pollen allergic patients with the allergen extracts. CONCLUSIONS: Commercial birch pollen extracts exhibit a considerable variability regarding allergen contents and hence deliver varying in vivo test results. These problems might be overcome with recombinant allergen-based preparations. PMID- 19302562 TI - Anti-inflammatory effects of 5-HT receptor antagonist, tropisetron on experimental colitis in rats. AB - BACKGROUND: There is a pressing need for research that will lead to the development of new therapeutic approaches for treating inflammatory bowel disease (IBD). The aim of this study was to investigate the effects of tropisetron, a 5 Hydroxytryptamine (5-HT)-3 receptor antagonist with anti-inflammatory properties in a model of experimental colitis in rat. MATERIALS AND METHODS: Acetic acid model of colitis in rats was used. Colitis was induced by intracolonal instillation of 4% (v/v) acetic acid. One hour after induction of colitis, intraperitoneal (IP) or intrarectal (IR) tropisetron (2 mg kg(-1), either route) or dexamethasone (1 mg kg(-1), either route) was administered. The severity of colitis was assessed 24 h later using macroscopic and microscopic changes of damaged colon, measurement of inflammatory cytokines interleukin-1beta, interleukin-6 and tumour necrosis factor-alpha levels and oxidative stress markers myeloperoxidase (MPO) and malondialdehyde (MDA) in colonic tissues. RESULTS: Tropisetron decreased colonic macroscopic and microscopic damage scores. This was associated with significant reduction in both neutrophil infiltration indicated by decreased colonic MPO activity and lipid peroxidation measured by MDA content, as well as a decreased colonic inflammatory cytokines. IR tropisetron decreased colonic damage that was associated with decreased neutrophil infiltration, lipid peroxidation and colonic inflammatory cytokines. Beneficial effects of tropisetron were lower than those of dexamethasone. No significant differences were observed between IP and IR administration with the exception of MDA level more diminished by IP tropisetron and dexamethasone. CONCLUSIONS: Tropisetron exert beneficial effects in experimental rat colitis and therefore might be useful in the treatment of IBD. PMID- 19302563 TI - Effects of hypocaloric very-low-carbohydrate diet vs. Mediterranean diet on endothelial function in obese women*. AB - Obesity is a cardiovascular risk factor associated with endothelial dysfunction, but the effect of different weight loss strategies on endothelial function is not known. The effect of diet on endothelial function in two hypocaloric diets, a very-low-carbohydrate diet (A) and a Mediterranean diet (M), was measured by brachial artery flow-mediated dilation (FMD). Design Using a longitudinal, randomized, open study design, subjects were engaged in a 2-month weight loss diet. FMD, inflammatory cytokines [interleukin-6 (IL-6) and tumour necrosis factor-alpha] and a marker of oxidative stress [8-iso-prostaglandin F2alpha (8 iso-PGF2alpha)] were measured in subjects on three occasions: before initiating the diet (T0), after 5-7 days of dieting (T5) and after 2 months of dieting (T60). The very short- and medium-term time points were established to discriminate respectively the effect of the diet itself (T5) from that of weight loss (T60). Twenty overweight/obese but otherwise healthy women (BMI: 27-34.9 kg m(-2); age 30-50 years) completed the study. Results Group A lost more weight (mean +/- SEM; -7.6 +/- 0.8 kg) than group M (-4.9 +/- 0.6 kg, P = 0.014) at T60. The FMD was not significantly different between the two groups at T0 (group A: 12.2 +/- 2.9% vs. group B: 10.3 +/- 2.3%, P = ns). In group A, FMD was significantly reduced at T5 and returned to baseline at T60; in group M, FMD increased at T5 and returned to baseline at T60 (P = 0.007 for diet x time interaction). Serum concentrations of IL-6 and 8-iso-PGF2alpha were not significantly different between the two groups at T0 and increased significantly at T5 only in group A (P < 0.001 and P < 0.005 respectively). Conclusion As endothelial dysfunction is known to be associated with acute cardiovascular events, this study suggests that the cardiovascular risk might be increased in the first days of a very-low-carbohydrate diet. PMID- 19302564 TI - Cyclosporine therapy in dermatology. PMID- 19302565 TI - Low-dose combination therapy of severe digital ulcers in diffuse progressive systemic sclerosis with the endothelin-1 receptor antagonist bosentan and the phosphodiesterase V inhibitor sildenafil. AB - Digital ulcers in progressive systemic sclerosis (PSS) are often refractory to therapy. A frequently chronic aggressive course can lead to the loss of acral limbs involved. A 73-year-old woman developed a dramatic worsening of her ulcerations despite maximum conventional therapy. Switching therapy to bosentan and sildenafil, both in low-dose regimens, resulted for the first time in ten years in a complete healing of the ulcers. If substantiated in a series of patients, the additive and perhaps synergistic clinical benefits of combining bosentan and sildenafil may be a valuable option for the treatment of acral ulcers in PSS. PMID- 19302566 TI - The WHO code of practice on the international recruitment of health personnel: we have only just begun. PMID- 19302567 TI - Global health ethics for students. AB - As a result of increased interest in global health, more and more medical students and trainees from the 'developed world' are working and studying in the 'developing world'. However, while opportunities to do this important work increase, there has been insufficient development of ethical guidelines for students. It is often assumed that ethics training in developed world situations is applicable to health experiences globally. However, fundamental differences in both clinical and research settings necessitate an alternative paradigm of analysis. This article is intended for teachers who are responsible for preparing students prior to such experiences. A review of major ethical issues is presented, how they pertain to students, and a framework is outlined to help guide students in their work. PMID- 19302568 TI - Local attitudes, moral obligation, customary obedience and other cultural practices: their influence on the process of gaining informed consent for surgery in a tertiary institution in a developing country. AB - The process of obtaining informed consent in a teaching hospital in a developing country (e.g. Nigeria) is shaped by factors which, to the Western world, may be seen to be anti-autonomous: autonomy being one of the pillars of an ideal informed consent. However, the mix of cultural bioethics and local moral obligation in the face of communal tradition ensures a mutually acceptable informed consent process. Paternalism is indeed encouraged by the patients who prefer to see the doctor as all-powerful and all-knowing, and this is buttressed by the cultural practice of customary obedience to those 'above you': either in age or social rank. The local moral obligation reassures the patients that those in authority will always look after others placed in their care without recourse to lengthy discussions or signed documentation, while the communal traditions ensure that the designated head of a family unit has the honor and sole responsibility of assenting and consenting to an operation to be carried out on a younger, or female, member of the family. Indeed it is to only a few educated patients that the informed consent process is deemed a shield against litigation by the doctors. This paper later addresses the need for physicians to update their knowledge on the process of informed consent through the attendance of biomedical ethics courses, which should highlight socio-cultural practices that may make this process different from the Western concept, but perfectly acceptable in this setting. PMID- 19302570 TI - Lupus erythematosus with exclusive involvement of the acrosyringia. AB - Cutaneous lesions of lupus erythematosus (LE) show a broad spectrum of clinicopathologic features. Histopathologically, besides typical patterns such as interface dermatitis, perivascular lymphocytic infiltrate and dermal mucin deposits, an involvement of the eccrine structures, especially the acrosyringium, may be observed. We describe the case of a 21-year-old woman with a 4-year history of systemic LE, who presented with a 'butterfly' rash over the cheeks as well as erythematous macules on the arms and decollete. Biopsy from one lesion on the arm revealed interface changes, necrotic keratinocytes and exocytosis of lymphocytes restricted only to the regions of the acrosyringia. The epidermis between affected acrosyringia was normal with no hints of interface dermatitis. The eccrine glands and coils were not affected. In the dermis there were only sparse inflammatory infiltrates. Differential diagnoses such as erythema multiforme, drug eruption and lichen planus could be ruled out because of histopathologic features and clinical presentation. This is an example of a peculiar histopathological variant of cutaneous LE, characterized by exclusive involvement of the acrosyringia. The histopathologic features represent a pitfall in the diagnosis and can be correctly interpreted only upon correlation with clinical data. PMID- 19302571 TI - Histiocytic erythema multiforme. AB - Erythema multiforme is histologically characterized by liquefactive degeneration along the dermal-epidermal junction, necrotic keratinocytes and a lymphocytic infiltrate. We report a 10-year-old boy with recurrent erythema multiforme major of undetermined etiology with unusual histologic findings. A skin biopsy taken at day 2 of his eruption revealed histologic features otherwise characteristic of erythema multiforme, but mediated instead by a CD68-positive infiltrate, resembling cutaneous Kikuchi's disease. To the best of our knowledge this is the first reported case of 'histiocytic' erythema multiforme. PMID- 19302572 TI - Tattoo-associated keratoacanthomas: a series of 8 patients with 11 keratoacanthomas. AB - BACKGROUND: Keratoacanthoma is interpreted by many dermatopathologists in the United States as a form of squamous cell carcinoma that can spontaneously involute. Rare examples arising in tattoos have been reported in the literature. MATERIALS AND METHODS: We retrospectively reviewed all cases from our institution received between 2000 and 2008 for any that reported a tumor within a tattoo. RESULTS: We identified eight patients with keratoacanthomas that arose within tattoos. One of the patients had four separate keratoacanthomas arising within two separate tattoos. Red tattoo ink was associated with 82% of the keratoacanthomas. CONCLUSIONS: Keratoacanthomas are more common than previously reported in tattoos and are easily misinterpreted. The association with red tattoo ink suggests a form of hypersensitivity-associated with adnexal hyperplasia. Tattoo-associated squamous tumors with innocuous nuclei, infundibulocystic structures, adnexal hyperplasia, and signs of regression should be reported as keratoacanthomas rather than as variants of squamous cell carcinoma. PMID- 19302573 TI - Racial/ethnic variations in associations between socioeconomic factors and tooth loss. AB - OBJECTIVES: To compare the associations between socioeconomic factors and tooth loss among White, Black, and Mexican-American people. METHODS: Analyses were conducted on 16,821 adults, using data from the National Health and Nutrition Examination Survey-III. Age- and multivariate-adjusted negative binomial regressions were used to explore the relation of socioeconomic factors, region of residence, gender, and foreign birth with the number of missing teeth. Effect modification by race/ethnicity was assessed by the inclusion of interaction terms. RESULTS: In multivariate-adjusted analyses, non-Hispanic White people with 9-12 years of education exhibited 71% higher mean number of missing teeth than those with >12 years of education [incidence rate ratio (IRR) = 1.71, 95% confidence interval (CI): 1.52-1.92]. Education was unrelated to the number of teeth among non-Hispanic Black people (IRR = 1.16; 95% CI: 1.00-1.35) or Mexican Americans (IRR = 1.10, 95% CI: 0.93-1.31). The poorest White people exhibited 39% more missing teeth, on average, than the most affluent White people, but no association between poverty and number of teeth was observed among Black or Mexican-American people. CONCLUSIONS: The associations between socioeconomic factors and tooth loss vary across race/ethnicity. This suggests that the health benefits associated with high socioeconomic status are not equally shared across racial/ethnic groups. PMID- 19302574 TI - Enamel defects and dental caries in 9-year-old children living in fluoridated and nonfluoridated areas of Auckland, New Zealand. AB - OBJECTIVES: This epidemiological study aims to investigate the developmental enamel defects and dental caries among 9-year-old children resident in fluoridated and nonfluoridated regions in Auckland, New Zealand. METHODS: A stratified, two-stage random selection design where strata were defined by fluoridation status, school size, and school decile. After informed consent was obtained, parents completed oral health questionnaires and children underwent dental examinations at school clinics. RESULTS: 612 children from 38 schools participated in the study. Overall, 175 (29%) children had lived continuously in fluoridated areas, 149 (24%) had lived continuously in nonfluoridated areas, and 288 (47%) had resided intermittently in fluoridated areas. Diffuse opacities were present in 117 (19%) children and deciduous teeth dental caries was seen in 370 (60%) children. After adjustment for covariates, a strong dose-response relationship between diffuse opacity and fluoridation status was found, with children who lived continuously in fluoridated areas being 4.17 times as likely to have diffuse opacities as children who lived continuously in nonfluoridated areas (P < 0.001). Conversely, a strong protective dose-response relationship between caries experience and fluoridation status was seen, with children who lived continuously in fluoridated areas being 0.42 times as likely to have dental caries as children who lived continuously in nonfluoridated areas (P < 0.001). CONCLUSIONS: Reticulated water fluoridation in Auckland reduces the risk of dental caries but increases the risk of diffuse opacities in 9-year-old children. Guidelines and health-promotion strategies that enable children to minimize their risk to diffuse opacities yet reduce their risk of dental caries should be reviewed. PMID- 19302575 TI - A simplified dental examination for detection of cavitated carious lesions in school setting. AB - OBJECTIVE: The objective of the study was to compare a simplified dental examination involving the use of a wooden spatula and a toothpick (TS examination) with the conventional dental mirror and probe examination (MP examination) for detection of cavitated carious lesions in schoolchildren. METHODS: The study involved three groups of caries-positive schoolchildren aged 4 5, 9-10 and 13-14 years with 179, 188 and 202 children, respectively. All children were examined by a trained and calibrated examiner using a wooden spatula and a toothpick under natural light. After a week the same examiner examined the three groups of children by a dental mirror and a Community Periodontal Index (CPI) probe. 'Teeth' were considered as units of measurement for data analysis. The WHO recommended criteria (2) for decayed, missing and filled teeth were followed for recording dental caries on a specially designed recording form. The sensitivity, specificity, positive predictive value, negative predictive value, false-positive rate and false-negative rate of TS examination were calculated using MP examination as the standard method. RESULTS: The specificity of TS examination was well above 95% in three types of dentition. Although the sensitivity of this type of examination in deciduous and mixed dentitions was almost comparable with that of MP examination, it was the lowest for permanent dentition but still within the acceptable limits. CONCLUSION: The TS examination can provide an alternative to traditional MP examination to undertake regular check-ups of schoolchildren for dental caries. PMID- 19302576 TI - Oral status and its association with general quality of life in older independent living south-Brazilians. AB - BACKGROUND: There are few studies assessing the importance of oral status, particularly tooth loss, edentulism and satisfaction with mastication, on the perception of quality of life (qol) in general. The objective of this study was to evaluate if oral status was associated with lower ratings in the WHO Quality of Life Questionnaire-Brief Version (WHOQOL-BREF) domains. METHODS: In this cross sectional study, a random sample of 872 Southern-Brazilians aged 60 years or more was evaluated using a structured questionnaire to assess sociodemographic data, qol (WHOQOL-BREF), and depressive symptoms (Geriatric Depression Scale). Oral examinations assessing the number of teeth and saliva flow were performed. Additionally, participants were asked to rate their satisfaction with the ability to chew. Correlates of poorer qol (i.e. values of the WHOQOL-BREF < or = median) were assessed by means of multivariate logistic regressions. RESULTS: Poorer qol in the physical domain was associated with increasing age, income, living in rural areas, >2 chronic conditions, intake of medications, depressive symptoms, edentulism, and reporting difficulty for chewing food. Poorer qol in the psychological domain was associated with income, schooling, >2 chronic conditions, intake of medications, depressive symptoms, and difficulty for chewing. Poorer qol in the environment domain was associated with income, living in urban areas, depressive symptoms, and difficulty for chewing. Poorer qol in the social relations domain was associated with living in urban areas, depressive symptoms, and difficulty for chewing. CONCLUSION: In the studied population, oral status was associated with the perception of quality of life in general. PMID- 19302577 TI - Intracellular parasitism with Toxoplasma gondii stimulates mammalian-target-of rapamycin-dependent host cell growth despite impaired signalling to S6K1 and 4E BP1. AB - The Ser/Thr kinase mammalian-target-of-rapamycin (mTOR) is a central regulator of anabolism, growth and proliferation. We investigated the effects of Toxoplasma gondii on host mTOR signalling. Toxoplasma invasion of multiple cell types rapidly induced sustained mTOR activation that was restricted to infected cells, as determined by rapamycin-sensitive phosphorylation of ribosomal protein S6; however, phosphorylation of the growth-associated mTOR substrates 4E-BP1 and S6K1 was not detected. Infected cells still phosphorylated S6K1 and 4E-BP1 in response to insulin, although the S6K1 response was blunted. Parasite-induced S6 phosphorylation was independent of S6K1 and did not require activation of canonical mTOR-inducing pathways mediated by phosphatidylinositol 3-kinase-Akt and ERK. Host mTOR was localized in a vesicular pattern surrounding the parasitophorous vacuole, suggesting potential activation by phosphatidic acid in the vacuolar membrane. In spite of a failure to phosphorylate 4E-BP1 and S6K1, intracellular T. gondii triggered host cell cycle progression in an mTOR dependent manner and progression of infected cells displayed increased sensitivity to rapamycin. Moreover, normal cell growth was maintained during parasite-induced cell cycle progression, as indicated by total cellular S6 levels. The Toxoplasma-infected cell provides a unique example of non-canonical mTOR activation supporting growth that is independent of signalling through either S6K1 or 4E-BP1. PMID- 19302578 TI - Stimulation of cell motility and expression of late markers of differentiation in human oral keratinocytes by Candida albicans. AB - A hallmark of the mucosa of immunocompromized hosts in oral candidiasis is a hyperkeratinized region heavily colonized with fungi at the surface of the terminally differentiated epithelium. To gain insight into the processes important for promoting mucosal invasion by fungi, we characterized the response of keratinocytes to the presence of Candida albicans. Indirect immunofluorescence and kymographic analyses revealed a multifaceted keratinocyte response of OKF6/TERT-2 cells to C. albicans that consisted of: cytoskeletal reorganization within 3 h, motility and cell expansion with formation of E-cadherin-mediated cell-cell adhesions within 6 h, increased expression of late differentiation markers and decreased expression of calprotectin. The initial expansive phase was followed by dissolution of cell-cell adhesions and a decrease in cell size accompanied by loss of E-cadherin. The keratinocyte response depended on soluble factors associated with hyphal growth as demonstrated using the efg1Delta/efg1Delta, cap1Delta/cap1Delta, als3Delta/als3Delta, hwp1Delta/hwp1Deltaand sap4-6Delta/sap4-6Delta mutants and was not observed in the presence of the non-pathogenic yeast, Saccharomyces cerevisiae. These studies show the potential for C. albicans to manipulate the stratified epithelial cells to a state of differentiation that is more permissive of fungal colonization of oral tissue, which is likely to play an important role in the pathogenesis of candidiasis. PMID- 19302579 TI - A translocated protein of Bartonella henselae interferes with endocytic uptake of individual bacteria and triggers uptake of large bacterial aggregates via the invasome. AB - Bartonella henselae enters human endothelial cells (ECs) by two alternative routes: either by endocytosis, giving rise to Bartonella-containing vacuoles or by invasome-mediated internalization. Only the latter process depends on the type IV secretion system VirB/VirD4 and involves the formation of cell surface associated bacterial aggregates, which get engulfed by EC membranes in an F-actin dependent manner, eventually resulting in their complete internalization. Here, we report that among the VirB/VirD4-translocated effector proteins BepA-BepG only BepG is required for triggering invasome-mediated internalization. Expression of BepG in the Bep-deficient DeltabepA-G mutant restored invasome-mediated internalization. Likewise, ectopic expression of BepG in ECs also restored invasome-mediated internalization of the DeltabepA-G mutant, while no discernable cytoskeletal rearrangements were triggered in uninfected cells. Rather, BepG inhibited endocytic uptake of B. henselae into Bartonella-containing vacuoles and other endocytic processes, that is, invasin-mediated uptake of Yersinia enterocolitica and uptake of inert microspheres. BepG thus triggers invasome mediated internalization primarily by inhibiting bacterial endocytosis. Bacteria accumulating on the cell surface then induce locally the F-actin rearrangements characteristic for the invasome. These cytoskeletal changes encompass both the rearrangement of pre-existing F-actin fibres and the de novo polymerization of cortical F-actin in the periphery of the invasome by Rac1/Scar1/WAVE- and Cdc42/WASP-dependent pathways that involve the recruitment of the Arp2/3 complex. PMID- 19302580 TI - Can hormones contained in mothers' milk account for the beneficial effect of breast-feeding on obesity in children? AB - Nutrition and growth during infancy are an emerging issue because of their potential link to metabolic health disorders in later life. Moreover, prolonged breast-feeding appears to be associated with a lower risk of obesity than formula feeding. Human milk is a source of various hormones and growth factors, namely adipokines (leptin and adiponectin), ghrelin, resistin and obestatin, which are involved in food intake regulation and energy balance. These compounds are either not found in commercial milk formulas or their presence is still controversial. Diet-related differences during infancy in serum levels of factors involved in energy metabolism might explain anthropometric differences and also differences in dietary habits between breast-fed (BF) and formula-fed (FF) infants later in life, and may thus have long-term health consequences. In this context, the recent finding of higher leptin levels and lower ghrelin levels in BF than in FF infants suggests that differences in hormonal values together with different protein intake could account for the differences in growth between BF and FF infants both during infancy and later in life. In this review, we examine the data related to hormones contained in mothers' milk and their potential protective effect on subsequent obesity and metabolic-related disorders. PMID- 19302581 TI - Comparison of three doses of leuprolide acetate in the treatment of central precocious puberty: preliminary results. AB - OBJECTIVE: Depot luteinizing-hormone releasing hormone (LHRH) agonist have been widely used for the treatment of central precocious puberty (CPP), but the optimal doses to obtain hormonal suppression are still unknown, especially in patients with higher weights. The goal of our study was to compare the efficacy of three leuprolide acetate (LA) preparations, suppressing gonadotropin secretion in patients with CPP. DESIGN: In an open 12-month protocol, we evaluated LA 7.5 mg/month, 11.25 and 22.5 every 3 months. PATIENTS: Fourteen girls with CPP and weights over 30 kg. MEASUREMENTS: Clinical, radiological and laboratory follow up: GnRH test plus LH, FSH 40 min post analogue was performed periodically. RESULTS: Pretreatment basal and LHRH stimulated LH levels between groups were not different. Basal and LHRH stimulated LH levels decreased significantly between baseline and from 3 up to 12 months of therapy in all groups (P = 0.001). GnRH stimulated LH peak <2 IU/l, the main efficacy criterion was met in 80, 75 and 100% of the children at 6 months in the 7.5, 11.25, 22.5 mg doses respectively. By 12 months, 100% of patients had LH suppressed to <2 IU/l. CONCLUSIONS: These results affirm that 3-month injections may be a satisfactory alternative for the therapy of children with CPP to avoid monthly injections. In addition, suppression of LH occurs sooner in the 3-month 22.5 mg LA dose compared to the 3 month 11.5 mg; therefore, adequate dosing may be important for optimal outcome. Further investigation is needed in more patients over 30 kg, with longer treatment duration, and ultimately final height consideration. PMID- 19302582 TI - The natural history of surgically treated but radiotherapy-naive nonfunctioning pituitary adenomas. AB - BACKGROUND AND OBJECTIVES: Transsphenoidal surgery is indicated for patients with nonfunctioning pituitary adenomas (NFPAs) causing compressive symptoms. Previous studies attempting to define the rate of recurrence/regrowth of surgically treated but radiation-naive NFPAs were somewhat limited by selection bias and/or small numbers and/or lack of consistency of findings between studies. A better understanding of the natural history of this condition could allow stratification of recurrence risk and inform future management. We aimed to define the natural history of a large, mainly unselected cohort with surgically treated, radiotherapy (RT)-naive NFPAs and to try to identify predictors of recurrence/regrowth. DESIGN: Case-note analysis of all patients who underwent surgery for NFPA in our hospital between 1980 and 2006 was undertaken. Median follow-up was 5.7 (range 1-25) years. PATIENTS: A total of 212 patients were identified of which 159 were suitable for analysis. 93% did not receive post operative RT. MEASUREMENT: Post-operative recurrent/regrowth was defined by any increase in tumour remnant size on serial post-operative pituitary imaging. RESULTS: Recurrence/regrowth was documented in 53 patients (33.5%). Multivariate analysis revealed size of the post-operative tumour remnant and length of follow up to be the two major determinants of recurrence/regrowth. The presence of a tumour with an extrasellar remnant was associated with the highest risk of recurrence (odds ratio 3.73 [CI: 1.97-7.09]), while no recurrence was seen in those with no residual tumour post-operatively and regrowth risk was intermediate for those with remaining intrasellar remnant. CONCLUSION: These results indicate that patients with post-operative tumour with an extrasellar remnant should be considered routinely for adjuvant RT to reduce the risk of tumour regrowth while those with no residual tumour can be safely observed. Individualized decisions should be made for patients with an intrasellar remnant. PMID- 19302583 TI - Measurement of salivary cortisol with liquid chromatography-tandem mass spectrometry in patients undergoing dynamic endocrine testing. AB - OBJECTIVE: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) eliminates cross-reactivity, which is a major limitation of immunoassays used for the measurement of salivary cortisol (SalC). We aimed to evaluate the potential of SalC measured by LC-MS/MS in patients undergoing assessment of the HPA axis. DESIGN AND PATIENTS: Cross-sectional study of 78 patients admitted for routine testing in a specialized endocrine unit. MEASUREMENTS: Matched serum and saliva samples were collected from 68 patients who had a short synacthen test (SST, 250 mcg im) and 10 patients who had an insulin tolerance test (ITT, insulin 0.15 U/kg iv). Serum cortisol (SerC) was measured with an automated immunoassay and SalC with LC-MS/MS. Adequate SerC responses were >500 nmol/l. RESULTS: In all patients with adequate responses, the relative increase in SalC was significantly higher than that in SerC [6.4(0.3-26.1) vs. 1.0(0.3-4.9), P < 0.0001)]. The SerC-SalC relationship was better explained by an exponential rather than a linear model (R(2)=0.83 vs. R(2)=0.65, both P < 0.0001). Based on 59 patients with adequate SerC responses to an SST, an adequate SalC response was defined as 8.3 nmol/l. Seven patients following an SST and three patients following an ITT showed inadequate responses in both SerC and SalC, but two patients with CBG deficiency showed a low SerC with normal SalC. CONCLUSIONS: We have shown an excellent diagnostic sensitivity and specificity of LC-MS/MS SalC in the assessment of the HPA axis and superiority over SerC when CBG levels are altered. The exponential relationship between SerC and SalC supports the concept of CBG binding capacity saturation. PMID- 19302584 TI - Unusual mid-shaft fractures during long-term bisphosphonate therapy. AB - BACKGROUND: Bisphosphonates are the most commonly prescribed medications for the treatment of osteoporosis. Although existing evidence supports a good safety profile, there is concern that chronic administration of these agents could result in severe suppression of bone turnover with increased risk of nonvertebral fractures. OBJECTIVE: The objective of this study was to report the clinical presentation, selected bone histomorphometry and X-ray images of patients who developed mid-shaft long bone fractures during bisphosphonate therapy, six of whom had bone biopsy for histomorphometery. RESULTS: Of the 13 patients who sustained atraumatic mid-shaft fractures, 10 were on alendronate and three were on risedronate therapy before the fractures. In addition to bisphosphonates, three patients were on oestrogen and two on tamoxifen concomitantly. Four patients with glucocorticoid-induced osteoporosis were on alendronate for 3-11 years along with glucocorticoid therapy. Bone histomorphometry showed severe suppression of bone turnover in five patients and low bone turnover in one patient. CONCLUSION: Long-term bisphosphonate therapy may increase the risk of unusual long bone mid-shaft fractures. This is probably due to prolonged suppression of bone turnover, which could lead to accumulation of microdamage and development of hypermineralized bone. At present, the scope of this complication in the larger context of patients receiving bisphosphonate therapy remains unknown, but appears to be small. PMID- 19302585 TI - All-trans retinoic acid-induced scrotal ulcer in a patient with acute promyelocytic leukaemia. PMID- 19302586 TI - Effective treatment of angiosarcoma on the nose by combination treatment with electron beam irradiation, recombinant interleukin-2 and docetaxel. PMID- 19302587 TI - Treatment of segmental cafe-au-lait macules using 1064-nm Q-switched Nd:YAG laser with low pulse energy. PMID- 19302588 TI - Bilateral osteochondritis dissecans-like osteochondral lesions in two patients: could isotretinoin be a cause? PMID- 19302589 TI - Amiloride derivatives induce apoptosis by depleting ER Ca(2+) stores in vascular endothelial cells. AB - BACKGROUND AND PURPOSE: Amiloride derivatives are blockers of the Na(+)/H(+) exchanger (NHE) and at micromolar concentrations have protective effects on cardiac and brain ischaemia/reperfusion injury but at higher concentrations also induce apoptosis. Here, we aimed to elucidate the mechanism related to this cytotoxic action. EXPERIMENTAL APPROACH: We quantified the expression of genes associated with endoplasmic reticulum (ER) stress and measured changes in luminal ER Ca(2+) concentration ([Ca(2+)](ER)) with a 'cameleon' indicator, D1ER. KEY RESULTS: Amiloride derivatives induced apoptosis in vascular endothelial cells, an effect that increased at alkaline extracellular pH. The potency order for cytotoxicity was 5-(N,N-hexamethylene)-amiloride (HMA) > 5-(N-methyl-N-isobutyl) amiloride > 5-(N-ethyl-N-isopropyl) amiloride (EIPA) >> amiloride. HMA dose dependently increased the transcription of the ER stress genes GADD153 and GADD34 and rapidly depleted [Ca(2+)](ER), mimicking the effects of the sarco/endoplasmic reticulum ATPase (SERCA) inhibitor thapsigargin. The NHE1-specific inhibitor HOE 694 inhibited NHE activity by 87% but did not alter [Ca(2+)](ER). The decrease in [Ca(2+)](ER) evoked by amiloride derivatives was also observed in HeLa cells and was mirrored by an increase in cytosolic Ca(2+) concentration. CONCLUSIONS AND IMPLICATIONS: Amiloride derivatives disrupt ER and cytosolic Ca(2+) homeostasis by a mechanism unrelated to NHE inhibition, most likely by interfering with the activity of SERCA. We propose that ER Ca(2+) depletion and subsequent ER stress provide a rationale framework for the apoptotic effects of amiloride derivatives. PMID- 19302590 TI - Endothelial Ca+-activated K+ channels in normal and impaired EDHF-dilator responses--relevance to cardiovascular pathologies and drug discovery. AB - The arterial endothelium critically contributes to blood pressure control by releasing vasodilating autacoids such as nitric oxide, prostacyclin and a third factor or pathway termed 'endothelium-derived hyperpolarizing factor' (EDHF). The nature of EDHF and EDHF-signalling pathways is not fully understood yet. However, endothelial hyperpolarization mediated by the Ca(2+)-activated K(+) channels (K(Ca)) has been suggested to play a critical role in initializing EDHF-dilator responses in conduit and resistance-sized arteries of many species including humans. Endothelial K(Ca) currents are mediated by the two K(Ca) subtypes, intermediate-conductance K(Ca) (KCa3.1) (also known as, a.k.a. IK(Ca)) and small conductance K(Ca) type 3 (KCa2.3) (a.k.a. SK(Ca)). In this review, we summarize current knowledge about endothelial KCa3.1 and KCa2.3 channels, their molecular and pharmacological properties and their specific roles in endothelial function and, particularly, in the EDHF-dilator response. In addition we focus on recent experimental evidences derived from KCa3.1- and/or KCa2.3-deficient mice that exhibit severe defects in EDHF signalling and elevated blood pressures, thus highlighting the importance of the KCa3.1/KCa2.3-EDHF-dilator system for blood pressure control. Moreover, we outline differential and overlapping roles of KCa3.1 and KCa2.3 for EDHF signalling as well as for nitric oxide synthesis and discuss recent evidence for a heterogeneous (sub) cellular distribution of KCa3.1 (at endothelial projections towards the smooth muscle) and KCa2.3 (at inter endothelial borders and caveolae), which may explain their distinct roles for endothelial function. Finally, we summarize the interrelations of altered KCa3.1/KCa2.3 and EDHF system impairments with cardiovascular disease states such as hypertension, diabetes, dyslipidemia and atherosclerosis and discuss the therapeutic potential of KCa3.1/KCa2.3 openers as novel types of blood pressure lowering drugs. PMID- 19302591 TI - Myoendothelial coupling in the mesenteric arterial bed; segmental differences and interplay between nitric oxide and endothelin-1. AB - BACKGROUND AND PURPOSE: We tested the hypothesis that activated arterial smooth muscle (ASM) stimulates endothelial vasomotor influences via gap junctions and that the significance of this myoendothelial coupling increases with decreasing arterial diameter. EXPERIMENTAL APPROACH: From WKY rats, first-, second-, third- and fourth-order branches of the superior mesenteric artery (MA1, MA2, MA3 and MA4 respectively) were isolated and mounted in wire-myographs to record vasomotor responses to 0.16-20 micromol x L(-1) phenylephrine. KEY RESULTS: Removal of endothelium increased the sensitivity (pEC(50)) to phenylephrine in all arteries. The nitric oxide (NO) synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) (100 micromol x L(-1)) did not modify pEC(50) to phenylephrine in all denuded arteries, and increased it in intact MA1, MA2 and MA3 to the same extent as denudation. However, in intact MA4, the effect of L-NAME was significantly larger (DeltapEC(50) 0.57 +/- 0.02) than the effect of endothelium removal (DeltapEC(50) 0.20 +/- 0.06). This endothelium-dependent effect of L-NAME in MA4 was inhibited by (i) steroidal and peptidergic uncouplers of gap junctions; (ii) a low concentration of the NO donor sodium nitroprusside; and (iii) by the endothelin-receptor antagonist bosentan. It was also observed during contractions induced by (i) calcium channel activation (BayK 8644, 0.001-1 micromol x L(-1)); (ii) depolarization (10-40 mmol x L(-1) K(+)); and (iii) sympathetic nerve stimulation (0.25-32 Hz). CONCLUSIONS AND IMPLICATIONS: These pharmacological observations indicated feedback control by endothelium of ASM reactivity involving gap junctions and a balance between endothelium-derived NO and endothelin-1. This myoendothelial coupling was most prominent in distal resistance arteries. PMID- 19302592 TI - Inhibition of anti-IgE mediated human mast cell activation by NO donors is dependent on their NO release kinetics. AB - BACKGROUND AND PURPOSE: Although the mast cell is a source of nitric oxide (NO), the effect of NO on human mast cells has not been defined. This study investigated if exogenous NO could affect human mast cell activation. EXPERIMENTAL APPROACH: Effects of different NO donors on immunoglobulin E (IgE) dependent activation of human-cultured mast cells (HCMC) derived from precursors in buffy coat were investigated by measuring histamine release. Intracellular NO in HCMC was monitored with confocal microscopy using the fluorescent NO indicator 4-amino-5-methylamino-2', 7'-difluorofluorescein. KEY RESULTS: Diethylamine NONOate (DEA/NO) and MAHMA NONOate (NOC-9), both have rapid NO release rates, only inhibited anti-IgE-induced histamine release when added to HCMC at the time of activation. NO donors with slower NO release kinetics were ineffective even after 30 min incubation. Confocal microscopy revealed that the effectiveness of NO donors was dependent on the availability of adequate NO inside HCMC during activation. The inhibitory action of DEA/NO was diminished by the NO scavenger, 2 (4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-3-oxide-1-oxyl but potentiated by the anti-oxidant, N-acetylcysteine (NAC). Furthermore, co-incubation with NAC allowed previously ineffective NO donors to suppress HCMC activation and thus suggested that NAC could increase the availability of NO from NO donors. CONCLUSIONS AND IMPLICATIONS: Our results demonstrated that NO was able to modulate human mast cell activation but only when enough NO was present at the time of cell activation. Our findings explain the controversy over the effectiveness of NO on mast cell degranulation and supports the possibility that NO donors could be beneficial for treating allergic inflammation. PMID- 19302593 TI - Anti-angiogenic and vascular disrupting effects of C9, a new microtubule depolymerizing agent. AB - BACKGROUND AND PURPOSE: The critical role of blood supply in the growth of solid tumours makes blood vessels an ideal target for anti-tumour drug discovery. The anti-angiogenic and vascular disrupting activities of C9, a newly synthesized microtubule-depolymerizing agent, were investigated with several in vitro and in vivo models. Possible mechanisms involved in its activity were also assessed. EXPERIMENTAL APPROACH: Microtubule-depolymerizing actions were assessed by surface plasmon resonance binding, competitive inhibition and cytoskeleton immunofluorescence. Anti-angiogenic and vascular disrupting activities were tested on proliferation, migration, tube formation with human umbilical vein endothelial cells, and in rat aortic ring, chick chorioallantoic membrane and Matrigel plug assays. Western blots and Rho activation assays were employed to examine the role of Raf-MEK-ERK (mitogen-activated ERK kinase, extracellular signal-regulated kinase) and Rho/Rho kinase signalling. KEY RESULTS: C9 inhibited proliferation, migration and tube formation of endothelial cells and inhibited angiogenesis in aortic ring and chick chorioallantoic membrane assays. C9 induced disassembly of microtubules in endothelial cells and down-regulated Raf-MEK-ERK signalling activated by pro-angiogenic factors. In addition, C9 disrupted capillary-like networks and newly formed vessels in vitro and rapidly decreased perfusion of neovasculature in vivo. Endothelial cell contraction and membrane blebbing induced by C9 in neovasculature was dependent on the Rho/Rho kinase pathway. CONCLUSIONS AND IMPLICATIONS: Anti-angiogenic and vascular disruption by C9 was associated with changes in morphology and function of endothelial cells, involving the Raf-MEK-ERK and Rho/Rho kinase signalling pathways. These findings strongly suggest that C9 is a new microtubule-binding agent that could effectively target tumour vasculature. PMID- 19302594 TI - Vascular calcification and secondary hyperparathyroidism of severe chronic kidney disease and its relation to serum phosphate and calcium levels. AB - BACKGROUND AND PURPOSE: Various complications consequent on disordered calcium and phosphate homeostasis occur frequently in chronic kidney disease (CKD) patients. Particularly, vascular calcification has high morbidity and mortality rates. There is a clear need for a better CKD model to examine various aspects of this disordered homeostasis. EXPERIMENTAL APPROACH: Oral dosing with adenine induced CKD in rats in only 10 days. Serum calcium, phosphate and parathyroid hormone were measured and calcification in aorta was assessed histologically. The effects of varying phosphorus content of diet or treatment with phosphate binders or active vitamin D(3) on these parameters were examined. KEY RESULTS: After adenine dosing, significant hyperphosphatemia, hypocalcemia and secondary hyperparathyroidism (2HPT) were observed during the experimental period of 15 weeks. Aortic calcification was detected in only some of the animals even at 15 weeks (approximately 40%). Treatment with vitamin D(3) for 18 days, even at a low dose (100 ng x kg(-1), 3-4 times week(-1), p.o), caused aortic calcification in all animals and increases in serum calcium levels up to the normal range. The vitamin D(3)-induced calcification was significantly inhibited by phosphate binders which lowered serum phosphate levels and the calcium x phosphate product, although serum calcium levels were elevated. CONCLUSIONS: These data suggest that rats dosed orally with adenine provide a more useful model for analysing calcium/phosphate homeostasis in severe CKD. Controlling serum calcium/phosphate levels with phosphate binders may be better than vitamin D(3) treatment in hyperphosphatemia and 2HPT, to avoid vascular calcification. PMID- 19302596 TI - Abstracts of the Thoracic Society of Australia & New Zealand and the Australian & New Zealand Society of Respiratory Science 2009 Annual Scientific Meetings. April 3-8, 2009. Darwin, Australia. PMID- 19302595 TI - Genistein aglycone reverses glucocorticoid-induced osteoporosis and increases bone breaking strength in rats: a comparative study with alendronate. AB - BACKGROUND AND PURPOSE: Glucocorticoid-induced osteoporosis (GIO) is the leading cause of secondary osteoporosis. Clinical evidence suggests a role for genistein aglycone in the treatment of post-menopausal osteopenia although proof of efficacy in comparison with currently available treatments is still lacking. To clarify this issue, we investigated the effects of genistein on bone compared with alendronate in experimental GIO. EXPERIMENTAL APPROACH: A total of 28 female Sprague-Dawley rats were used. GIO was induced by daily injections of methylprednisolone (MP; 30 mg x kg(-1) s.c.) for 60 days. Sham GIO animals (Sham MP) were injected daily with the MP vehicle. At the end of the osteoporosis development period, MP rats were randomized to receive: vehicle (n= 7), genistein aglycone (5 mg x kg(-1) s.c.; n= 7) or alendronate (0.03 mg x kg(-1) s.c.; n= 7). Treatment lasted 60 days. Sham-MP animals were treated with vehicle for an additional 60 days. At the beginning and at the end of treatments, animals were examined for bone mineral density and bone mineral content. Bone-alkaline phosphatase and carboxy-terminal collagen cross links were determined; femurs were removed and tested for breaking strength and histology. KEY RESULTS: Genistein aglycone showed a greater increase in bone mineral density, bone mineral content and in breaking strength than alendronate and significantly increased bone-alkaline phosphatase (bone formation marker), reduced carboxy terminal collagen cross links (bone resorption marker), compared with alendronate. Both treatments improved bone histology and the histological score. CONCLUSION AND IMPLICATIONS: The results strongly suggest that the genistein aglycone might be an alternative therapy for the management of secondary osteoporosis. PMID- 19302631 TI - Paradigm shift to address drug resistant tuberculosis in people living with HIV needed, and needed now. PMID- 19302632 TI - Evaluation of the Kudoh swab method for the culturing of Mycobacterium tuberculosis in rural areas. AB - OBJECTIVE: To compare the simple, swab 'Kudoh method' for culturing Myobacterium tuberculosis from sputum samples, to the standard Petroff digestion decontamination procedure. The Kudoh method, which requires no centrifugation and takes only 4-5 min per sample, was also evaluated for its performance in a rural setting. METHODS: Two hospital laboratories in Caracas, Venezuela processed 314 sputum samples, in parallel, with both methods. Separately, sputum specimens were cultured with the Kudoh swab method in a field environment with minimal laboratory facilities. RESULTS: In the hospital laboratories, the sensitivity of the Kudoh swab method was comparable to that of the standard Petroff culture procedure. The swab method also performed satisfactorily in the field, improving the diagnostic sensitivity by 21% over microscopic examination alone. CONCLUSION: The Kudoh swab method is an acceptable alternative for culturing mycobacteria that is particularly suitable for rural laboratories lacking adequate infrastructure for the Petroff method. PMID- 19302633 TI - British Consensus Guidelines on Intravenous Fluid Therapy for Adult Surgical Patients (GIFTASUP): Cassandra's view. PMID- 19302634 TI - Effects of moderate acute isovolaemic haemodilution on myocardial function in patients undergoing coronary surgery under volatile inhalational anaesthesia. AB - When myocardial oxygen demand is increased by elevated heart rate in patients undergoing coronary artery surgery under total intravenous anaesthesia, acute isovolaemic haemodilution may be associated with a deterioration of cardiac function. We investigated the effects of acute isovolaemic haemodilution during volatile inhalational anaesthesia. Forty patients undergoing coronary surgery were randomly assigned to two groups according to the rate of atrioventricular pacing (Group 70 at 70.min(-1) and Group 90 at 90.min(-1)). While paced at the fixed heart rate, acute isovolaemic haemodilution was performed before the start of cardiopulmonary bypass. In both groups mean (SD) stroke volume increased with haemodilution (from 65 (9) to 83 (10) ml.min(-1) (p < 0.01) in Group 70 and from 65 (9) to 81 (9) ml.min(-1) (p < 0.01) in Group 90) as a result of a decrease in systemic vascular resistance (from 1175 (231) to 869 (164) dynes.s.cm(-5) (p < 0.01) and from 1060 (185) to 849 (146) dynes.s.cm(-5) (p < 0.01), respectively) and an increase in end-diastolic volume (from 1049 (234) to 1405 (211) ml (p < 0.01) and from 1078 (106) to 1438 (246) ml (p < 0.01), respectively). Left ventricular pressure-derived data remained unchanged with acute isovolaemic haemodilution in both groups. PMID- 19302635 TI - The effect of esmolol on the QTc interval during induction of anaesthesia in patients with coronary artery disease. AB - The aim of this study was to evaluate whether esmolol has an effect on QT interval during induction of anaesthesia using etomidate and fentanyl in patients with known coronary artery disease. Sixty patients were prospectively randomised to either a control group or the esmolol group. Esmolol was administered as a bolus 1 mg.kg(-1), followed by a continuous infusion at 250 microg.kg(-1)min(-1). All patients received etomidate 0.3 mg.kg(-1) and fentanyl 15 microg.kg(-1). The ECG was recorded prior to induction of anaesthesia (T0), 5 min following the start of drug infusions (T1), 1 min following etomidate (T2), 3 min following vecuronium (T3), 30 s (T4), 2 min (T5) and 4 min (T6) after intubation. In the esmolol group, QTc interval was significantly shorter at T1, T2 and T4 compared to the control group (p < 0.05). In conclusion, QTc interval increased following tracheal intubation during induction of anaesthesia using etomidate and fentanyl. An infusion of Esmolol attenuated the QTc interval prolongation associated with tracheal intubation. PMID- 19302636 TI - The relationship between cerebral and somatic oxygenation and superior and inferior vena cava flow, arterial oxygenation and pressure in infants during cardiopulmonary bypass. AB - We investigated blood flow and regional oxygenation (rSO(2)) during cardiopulmonary bypass (CPB). Twenty infants (mean (SD) age 5 (3) months, weight 5.4 (1.6) kg) were prospectively studied. Total CPB and superior vena cava (SVC) flow were measured using Transonic Bypass Flowmeters, inferior vena cava (IVC) flow derived arithmetically and rSO(2) measured using Near Infra-Red Spectroscopy. Mean SVC flow was 51.3 (14.8) ml.kg(-1).min(-1) and mean IVC flow 62.5 (19.0) ml.kg(-1).min(-1). Mean cerebral rSO(2) was 71 (11)% and somatic rSO(2) 55 (13)%. Cerebral and somatic rSO(2) showed no correlation with SVC and IVC flow. Cerebral rSO(2) showed a positive correlation with P(a)co(2), mean arterial pressure (MAP) and haematocrit (p < 0.0001). Somatic rSO(2) showed a positive correlation with MAP and haematocrit (p = 0.01, p = 0.02). In conclusion, the distribution of blood flow during CPB varies. The most important factor affecting this is P(a)CO(2). Cerebral and somatic oxygenation are unaffected by flow but significantly influenced by MAP, haematocrit and P(a)CO(2). PMID- 19302637 TI - The efficacy of a smoking cessation programme in patients undergoing elective surgery: a randomised clinical trial. AB - It is known that smokers constitute an important risk group of patients undergoing surgery. It is unknown how smoking cessation intervention initiated 4 weeks prior to elective surgery affects the probability of permanent cessation. We randomly assigned 117 patients, scheduled to undergo elective orthopaedic and general surgery, to smoking cessation intervention and control group. The intervention group underwent a programme initiated, on average, 4 weeks prior to surgery with weekly meetings or telephone counselling and were provided with free nicotine replacement therapy (NRT). The control group received standard care. As a result, 20/55 (36%) patients the intervention group vs 1/62 (2%) in the control group became completely abstinent throughout the peri-operative period (p < 0.001). After 1 year, those in the intervention group was most likely to be abstinent (18/55 (33%) vs 9/62 (15%) of the controls (p = 0.03). Level of nicotine dependence and obesity seemed to be a predictor of long-term abstinence (p = 0.02). PMID- 19302638 TI - Ambulation in labour and delivery mode: a randomised controlled trial of high dose vs mobile epidural analgesia. AB - Compared to high-dose epidurals where mobility is impossible, mobile epidurals have been shown to reduce instrumental vaginal delivery rates. The mechanism for this benefit may depend on women walking or adopting upright postures during labour. We investigated maternal motor power and ambulation of 1052 primparous women randomised to high-dose epidural (Control), Combined Spinal Epidural (CSE) or Low-Dose Infusion (LDI) as a pre-specified, secondary outcome of the Comparative Obstetric Mobile Epidural Trial. Modified Bromage power scores and the level of mobility a woman actually achieved were recorded each hour after epidural placement during first and second stage, until delivery. Relative to control, significantly more women maintained normal leg power throughout labour in both mobile groups and significantly more women with CSE maintained superior leg power for longer than with LDI. Observational analysis did not demonstrate an association between the level of ambulation a woman actually achieved after epidural placement and delivery mode. PMID- 19302639 TI - Effect of epidural saline washout on regression of sensory and motor block after epidural anaesthesia with 2% lidocaine and fentanyl in elderly patients. AB - Seventy elderly males received lumbar epidural anaesthesia with 12 ml of 2% lidocaine containing fentanyl 50 mug. At the end of transurethral surgery, the washout group (n = 33) received an epidural bolus of 30 ml saline while the control group (n = 34) did not. Mean (SD) times to 1-grade (17.2 (11.9) vs 32.7 (11.3) min) and 2-grade regression (23.8 (12.2) vs 56.0 (23.9) min) of motor block, 3-dermatomal sensory regression (31.4 (11.6) vs 42.2 (14.4) min for cold and 30.8 (15.6) vs 40.6 (14.2) min for pinprick), and regression to S1 (57.7 (16.1) vs 76.2 (20.2) min for cold and 56.8 (17.3) vs 69.2 (16.2) min for pinprick) were significantly shorter in the washout group than the control group. There were no differences in postoperative pain scores and side effects between the two groups. We concluded that epidural washout facilitates regression of both motor and sensory block following epidural anaesthesia without reducing the postoperative analgesic benefit. PMID- 19302640 TI - A prospective study of the time to evacuate acute subdural and extradural haematomas. AB - We performed a prospective, single-centre study of times to treatment of patients with life-threatening, traumatic, extra- and subdural haematomas requiring surgical evacuation between May 2006 and May 2007. The mean time to surgical decompression was 5.0 h and 32% were performed within 4 h. Patients who initially presented to a district hospital and required transfer for neurosurgery were decompressed in 5.4 h vs 3.7 h for those admitted directly. The current standard of surgical evacuation of all haematomas within 4 h is not being met. Delays were identified in every stage in the management of these patients and no single step was identified as the major cause. Initial treatment in district hospitals led to delays greater than the added driving time. There may be time savings from carrying out treatment steps in parallel instead of in series. PMID- 19302641 TI - Ketamine or alfentanil administration prior to propofol anaesthesia: the effects on ProSeal laryngeal mask airway insertion conditions and haemodynamic changes in children. AB - This study was designed to compare the effects of ketamine and alfentanil administered prior to induction of anaesthesia with propofol, on the haemodynamic changes and ProSeal laryngeal mask airway (PLMA) insertion conditions in children. Eighty children, aged between 3-132 months, were randomly allocated to receive either alfentanil 20 microg.kg(-1) (alfentanil group) or ketamine 0.5 mg.kg(-1) (ketamine group) before induction of anaesthesia. Ninety seconds following the administration of propofol 4 mg.kg(-1), a PLMA was inserted. In the ketamine group, heart rate and mean arterial pressure were higher during the study period compared with the alfentanil group (p < 0.05). The time for the return of spontaneous ventilation was prolonged in the alfentanil group (p = 0.004). In conclusion, we found that the administration of ketamine 0.5 mg.kg(-1) with propofol 4 mg.kg(-1) preserved haemodynamic stability, and reduced the time to the return of spontaneous ventilation, compared with alfentanil 20 microg.kg( 1) during PLMA placement. In addition, the conditions for insertion of the PLMA with ketamine were similar to those found with alfentanil. PMID- 19302642 TI - Performance of four carbon dioxide absorbents in experimental and clinical settings. AB - To evaluate the performance of four kinds of carbon dioxide (CO(2)) absorbents (Medisorb GE Healthcare, Amsorb Plus Armstrong Medical, YabashiLime Yabashi Industries, and Sodasorb LF Grace Performance Chemicals), we measured their dust production, acceptability of colour indicator, and CO(2) absorption capacity in in vitro experimental settings and the concentration of compound A in an inspired anaesthetic circuit during in vivo clinical practice. In vitro, the order of the dust amount was Sodasorb LF > Medisorb > Amsorb Plus = YabashiLime both before and after shaking. The order of the color acceptability was similar: Sodasorb LF > Amsorb Plus = Medisorb > YabashiLime both initially and 16 h after CO(2) exhaustion. During exposure to 200 ml.min(-1) CO(2) in vitro, the period until 1 kg of fresh soda lime allowed inspired CO(2) to increase to 0.7 kPa (as a mark of utilisation of the absorbent) was longer with Medisorb (1978 min) than with the other absorbents (1270-1375 min). In vivo, compound A (1.0% inspired sevoflurane) was detected only when using Medisorb. While Medisorb has the best ability to absorb CO(2), it alone produces compound A. PMID- 19302643 TI - Assessing the performance of a consultant anaesthetist by control chart methodology. AB - The performance of trainee anaesthetists has been assessed by control chart methodology, but while assessment of consultants' performance has been suggested, published reports are rare. In order to test the value of control charts, I monitored my insertion of arterial and central venous cannulae over 3 years, documenting success or failure both for the first pass of the needle and for the intended site of cannulation. The resultant control charts clearly show episodes of changed performance. It was necessary to modify the technique used to assess trainees, specifically with relation to definitions of success, failure and 'acceptable' failure rates. The information obtained from these charts is well suited to inform an appraisal or revalidation discussion with both reflective and assessment elements. PMID- 19302644 TI - A retrospective study of anaesthetic caseload of Specialist Registrars following the introduction of new working patterns in the Wessex region. AB - The impact of implementing the European Working Time Directive (EWTD) and the consequent reduction in hours of work has led to concerns about training. This retrospective study compared total caseload, obstetric caseload, out-of-hours caseload and supervision levels of trainee anaesthetists in one region of the UK (Wessex) before and after the implementation of EWTD compliant rotas. Anaesthetic trainee logbooks submitted at the annual Record of In-Training Assessment were compared between the years 1999 and 2006. The logbook data was divided into two groups, group A (13-h shift) and group B (24-h on-call rota). There was no difference in total caseload, obstetric caseload or supervision. Out-of-hours caseload was greater in group A (p < 0.01). PMID- 19302645 TI - Remifentanil in paediatric anaesthetic practice. AB - Remifentanil is a synthetic opioid, first introduced into clinical practice in 1996. Its unique pharmacokinetic profile has resulted in a gradual increase in its popularity in paediatric anaesthesia. It is an opioid of high potency and rapid clearance, consequently lacking problems of accumulation. These characteristics give it a high degree of predictability and it has become an attractive choice for a wide variety of anaesthetic challenges, from premature neonates to the elderly. Neonates and infants have a higher clearance than older children and, as a result, remifentanil has additional benefits in this group. Remifentanil can be described as the only consistently predictable opioid in paediatric practice. PMID- 19302646 TI - Evaluation of tracheal tube introducers in simulated difficult intubation. AB - In a randomised cross-over study, 72 anaesthetists attempted to place Pro Breathe, new Portex, and Frova single-use tracheal tube introducers and an Eschmann multiple-use introducer in the trachea of a manikin set to simulate a grade 3 laryngeal view. Successful placement (proportion, 95% confidence interval) of either the Frova (78%, 67-86%) or the Eschmann introducer (64%, 52 74%) was significantly more likely (p < 0.0001) than with the Pro-Breathe (4%, 1 12%) or the new Portex introducer (13%, 7-22%). The difference between the success rates for the Frova and the Eschmann introducers (p = 0.08) was not significant. A separate experiment revealed that the peak force that could be exerted by the Pro-Breathe, new Portex and Frova single-use introducers were three to six times greater than that which could be exerted by the Eschmann introducer (p < 0.0001). The single-use introducers are more likely to cause tissue trauma during placement, particularly if held close to the tip. PMID- 19302647 TI - Optimising tracheal intubation success rate using the Airtraq laryngoscope. AB - In this study we have used a video-recording, retrospective analysis technique to evaluate the influence of the Airtraq laryngoscope manipulations and the resulting changes in position of the glottic opening and inter-arytenoids cleft, on the success rate of tracheal intubation. The video recordings of the internal views of 109 tracheal intubation attempts, in 50 anaesthetised patients were analysed. We demonstrated that successful tracheal intubation using the Airtraq laryngoscope require the glottic opening to be centred in the view, and positioning the inter-arytenoid cleft medially below the horizontal line in the centre of the view. We also demonstrated that repositioning of the Airtraq laryngoscope following a failed tracheal intubation attempt required the performance of a standard series of manoeuvres. PMID- 19302648 TI - Forces required to remove bougies from tracheal tubes. AB - Following a critical incident on our labour ward, we investigated the forces required to remove different bougies from different tracheal tubes and the effect of lubrication on ease of removal. Two studies were conducted: firstly examining the differences between six different bougies with a standard tracheal tube, and secondly examining the differences between four different tracheal tubes with a standard bougie. The forces varied amongst both the different bougies (p < 0.0001) and the different tracheal tubes (p < 0.0001). Removal was generally easier with lubrication but when corrected for multiple comparisons this did not reach statistical significance. PMID- 19302649 TI - Comparison of the Glidescope and Airtraq optical laryngoscopes in patients undergoing direct microlaryngoscopy. AB - Optical laryngoscopes have been developed to facilitate difficult airway management. The Airtraq is a single-use device and the GlideScope is reusable. In this study, the Airtraq and the Glidescope were compared in 60 ASA I-III patients with tumours of the upper airway undergoing direct endoscopic microlaryngoscopy. Patients were randomly assigned to the Airtraq or the Glidescope group and the Cormack and Lehane grade was assessed by Macintosh laryngoscopy prior to tracheal intubation. There were no differences in tracheal intubation success rates or duration of intubation attempts between both devices. The Cormack and Lehane grade was improved in 77% and 82% of cases in the Airtraq and Glidescope group, respectively. Blood traces on the device and traumatic pharyngeal lesions were found more frequently in the Airtraq group. The Airtraq and Glidescope laryngoscopes are valuable tools for the management of patients with potentially difficult airways with the Glidescope appearing to be less traumatic. PMID- 19302650 TI - Re: Modelling patient flows to aid decision making for critical care capacities and organisation. PMID- 19302651 TI - A NICE idea or a high price to pay? Local assessment of national guidelines. PMID- 19302653 TI - Alternatives to glucose-insulin-potassium infusions. PMID- 19302654 TI - Target driven analgesia for total knee arthroplasty. PMID- 19302656 TI - Trauma: we care. More for discussion. PMID- 19302658 TI - Fault in Blease Frontline 860 anaesthetic machine. PMID- 19302660 TI - Primary hypopituitarism and peri-operative steroid supplementation. PMID- 19302661 TI - Blocked scavenging causing increased Fico(2). PMID- 19302662 TI - One millilitre could make all the difference! PMID- 19302663 TI - Phenytoin toxicity masquerading as motor neurone disease. PMID- 19302664 TI - Ultrasound guided location and removal of retained arterial cannula fragment. PMID- 19302673 TI - Repeatability of corneal and ocular aberration measurements and changes in aberrations over one week. AB - BACKGROUND: Both Shack-Hartmann aberrometry (IRX3, Imagine Eyes, Orsay, France) and Scheimpflug photography (Oculus Pentacam, Oculus Inc Wetzlar, Germany) are known to provide repeatable measurements. The variability in measurements of corneal and ocular aberrations obtained with these instruments over one week had not been assessed. The aim was to study the variability in corneal and ocular aberrations in the human eye over one week and to determine the impact of age on corneal and crystalline lens aberrations and on the variability of these measurements. METHODS: Monochromatic ocular aberrations were measured with the IRX3 and monochromatic corneal aberrations were measured using Scheimpflug photography on one eye of 23 normal subjects at periods of a few seconds, one hour and one week after the initial measurement. RESULTS: No significant differences were found between the initial aberrations and aberrations measured at 59 +/- 24 seconds, 1.10 +/- 0.24 hours and 7.11+/- 0.31 days later. Analysis with Bland and Altman plots showed that measurements with both instruments were highly repeatable over the times studied. There was no relationship between age and the variance of corneal and ocular aberrations (higher order, spherical and coma aberrations). Corneal spherical aberration did not show a significant correlation with age, whereas the lens aberrations changed from being negative in the younger age groups to positive in the older age group, however, these differences failed to reach statistical significance (p > 0.05). CONCLUSION: The variability found in all the measurements was small and not clinically significant and could be attributable to instrument noise, changes in the tear film and to small fixational eye movements. PMID- 19302674 TI - Contralateral comparison of wavefront-guided LASIK surgery with iris recognition versus without iris recognition using the MEL80 Excimer laser system. AB - PURPOSE: To compare outcomes in wavefront-guided LASIK performed with iris recognition software versus without iris recognition software in different eyes of the same patient. METHODS: A randomised, prospective study of 104 myopic eyes of 52 patients undergoing LASIK surgery with the MEL80 excimer laser system was performed. Iris recognition software was used in one eye of each patient (study group) and not used in the other eye (control group). Higher order aberrations (HOAs), contrast sensitivity, uncorrected vision (UCV), visual acuity (VA) and corneal topography were measured and recorded pre-operatively and at one month and three months post-operatively for each eye. RESULTS: The mean post-operative sphere and cylinder between groups was similar, however the post-operative angles of error (AE) by refraction were significantly smaller in the study group compared to the control group both in arithmetic and absolute means (p = 0.03, p = 0.01). The mean logMAR UCV was significantly better in the study group than in the control group at one month (p = 0.01). The mean logMAR VA was significantly better in the study group than in control group at both one and three months (p = 0.01, p = 0.03). In addition, mean trefoil, total third-order aberration, total fourth-order aberration and the total scotopic root-mean-square (RMS) HOAs were significantly less in the study group than those in the control group at the third (p = 0.01, p = 0.05, p = 0.04, p = 0.02). By three months, the contrast sensitivity had recovered in both groups but the study group performed better at 2.6, 4.2 and 6.6 cpd (cycles per degree) than the control group (p = 0.01, p < 0.01, p = 0.01). CONCLUSIONS: LASIK performed with iris recognition results in better VA, lower mean higher-order aberrations, lower refractive post-operative angles of error and better contrast sensitivity at three months post-operatively than LASIK performed without iris recognition. PMID- 19302675 TI - Describing ocular aberrations with wavefront vergence maps. AB - A common optometric problem is to specify the eye's ocular aberrations in terms of Zernike coefficients and to reduce that specification to a prescription for the optimum sphero-cylindrical correcting lens. The typical approach is first to reconstruct wavefront phase errors from measurements of wavefront slopes obtained by a wavefront aberrometer. This paper applies a new method to this clinical problem that does not require wavefront reconstruction. Instead, we base our analysis of axial wavefront vergence as inferred directly from wavefront slopes. The result is a wavefront vergence map that is similar to the axial power maps in corneal topography and hence has a potential to be favoured by clinicians. We use our new set of orthogonal Zernike slope polynomials to systematically analyse details of the vergence map analogous to Zernike analysis of wavefront maps. The result is a vector of slope coefficients that describe fundamental aberration components. Three different methods for reducing slope coefficients to a spherocylindrical prescription in power vector forms are compared and contrasted. When the original wavefront contains only second order aberrations, the vergence map is a function of meridian only and the power vectors from all three methods are identical. The differences in the methods begin to appear as we include higher order aberrations, in which case the wavefront vergence map is more complicated. Finally, we discuss the advantages and limitations of vergence map representation of ocular aberrations. PMID- 19302684 TI - Some difficulties in interpreting psychophysiological research with children. PMID- 19302676 TI - Marital conflict and children's externalizing behavior: interactions between parasympathetic and sympathetic nervous system activity. AB - Toward greater specificity in the prediction of externalizing problems in the context of interparental conflict, interactions between children's parasympathetic and sympathetic nervous system (PNS and SNS) activity were examined as moderators. PNS activity was indexed by respiratory sinus arrhythmia (RSA) and RSA reactivity (RSA-R) to lab challenges. SNS activity was indexed by skin conductance level (SCL) and SCL reactivity (SCL-R) to lab challenges. Moderation hypotheses were examined in 3 multi-informant studies with children ranging in age between 7 and 9 in Studies 1 and 2 and between 6 and 12 in Study 3. Findings are robust across studies and provide the first reported evidence of interactions between PNS and SNS activity as moderators of the association between children's exposure to marital conflict and externalizing behaviors. More specifically, opposing action of the PNS and SNS (i.e., coactivation and coinhibition) operated as a vulnerability factor for externalizing behavior in the context of marital conflict. Conversely, coordinated action of the PNS and SNS (i.e., reciprocal PNS or SNS activation) operated as a protective factor. Results are supportive of the authors' proposed biopsychosocial framework in which individual differences in the coordination of the activity of the PNS and SNS can function as vulnerability or protective factors in the context of family risk. Findings extend current theory indicating the importance of multisystem investigations for clarifying inconsistencies and discrepancies in the literature linking environmental stress, physiological responses, and child adjustment. PMID- 19302685 TI - When negative is positive. PMID- 19302688 TI - Family nurse practitioner clinical requirements: is the best recommendation 500 hours? AB - PURPOSE: The purpose of the article was to stimulate further dialog around the recommended number of clinical hours required for students in family nurse practitioner (FNP) programs. DATA SOURCES: Nursing literature was reviewed for research supporting the current clinical hour requirement. Both authors are faculty in FNP programs that provided experiential data for the article. CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: There are no scientific data upon which to base the number of required clinical hour. Researchers need to identify what type and how many clinical hours are needed to prepare competent new FNP graduates. PMID- 19302689 TI - Prevalence of cardiovascular disease and risk factors in a type 2 diabetic population of the North Catalonia diabetes study. AB - PURPOSE: The purpose of the study was to evaluate the prevalence of cardiovascular disease (CVD), cardiovascular risk factors (CVRFs), and their control in patients with type 2 diabetes mellitus (T2DM) at primary care settings from the North Catalonia Diabetes Study (NCDS). DATA SOURCES: In this multicentre cross-sectional descriptive study, data were collected from a random sample of 307 patients with T2DM. The prevalence of CVD, CVRF, metabolic syndrome (MS), coronary heart disease (CHD) risk at 10 years (Framingham Point Scores), and CVRF control was evaluated. MS and lipid profiles were established according to Adult Treatment Panel III criteria. CONCLUSIONS: CVD prevalence was 22.0% (CHD: 18.9% and peripheral ischemia: 4.5%) and more frequent in men. The prevalence of selected CVRF was: hypertension: 74.5%; dyslipidemia: 77.7%; smoking: 14.9%; obesity 44.9%, and familial CVD: 38.4%. Three or more CVRFs, including T2DM, were observed in 91.3%. MS prevalence was 68.7%. Framingham score was 10.0%, higher in men than in women. CVD prevalence was related to: age, number of CVRFs, duration of diabetes, familial history of CVD, waist circumference, hypertension, lipid profile, kidney disease, and Framingham score, but not to MS by itself. Correct lipid profiles and blood pressure were only observed in 18.9% and 24.0%, respectively, whereas platelet aggregation inhibitors were only recorded in 16.1% of the patient cohort. MS presence was not an independent risk factor of CVD in our study. IMPLICATIONS FOR PRACTICE: The high prevalence of CVD and an inadequate control of CVRF, which were apparent in the NCDS population, would suggest that advanced practice nurses should consider incorporating specific cardiovascular assessment in their routine care of persons with T2DM. PMID- 19302690 TI - Retinoblastoma: an overview for advanced practice nurses. AB - PURPOSE: To provide an overview of the current literature surrounding the detection and management of retinoblastoma as well as care for the long-term survivor to serve as a guide for advanced practice nurses. DATA SOURCES: Comprehensive review of the literature on retinoblastoma including case studies, journal reviews, retrospective studies, and briefs. CONCLUSIONS: Retinoblastoma is a pediatric intraocular malignancy usually detected in the primary care setting. Typically asymptomatic, primary care providers will often identify the presence of retinoblastoma upon routine eye exam. Prompt diagnosis and commencement of treatment are paramount as delay in diagnosis leads to higher morbidity and mortality. Treatment options vary depending on the extent of disease. IMPLICATIONS FOR PRACTICE: Although current survival rates for retinoblastoma exceed 90%, individual visual outcome and survival are dependent upon early detection and prompt referral. In addition to research and education of survivors and families, advanced practice nurses play a key role in early detection and maintaining the current survival rate. PMID- 19302691 TI - Latent autoimmune diabetes of adulthood (LADA): an often misdiagnosed type of diabetes mellitus. AB - PURPOSE: The purpose of this article is to raise awareness about a frequently misdiagnosed form of diabetes, latent autoimmune diabetes of adulthood (LADA), to describe its clinical and epidemiological characteristics, and to compare them to those of the more common and widely known types of diabetes, type 1 diabetes mellitus (DM) and type 2 DM. DATA SOURCES: A review of the pertinent literature describing the features of LADA from 2000-2007 is summarized. CONCLUSIONS: LADA is a rather common and often underrecognized form of diabetes whose clinical presentation falls somewhere between that of type 1 DM and type 2 DM. From a pathophysiological perspective, it is more closely related to type 1 DM, and some have even used the term type 1.5 diabetes to refer to it; however, it is most often misdiagnosed and treated as type 2 DM. IMPLICATIONS FOR PRACTICE: Nurse practitioners (NPs) should always consider alternate diagnoses when patients with newly or previously identified adult-onset diabetes mellitus do not fit the traditional stereotype of type 2 DM (i.e., overweight with signs of insulin resistance and a significant family history of diabetes). Statistically, strong consideration must be given to the diagnosis of LADA, especially in those who are of normal weight, show little evidence of insulin resistance, and have hardly any family history of diabetes. Knowing the patient's exact diabetes type can give the NP a much greater understanding of the natural history of the patient's disease, the changes that may occur as the patient ages, and how to optimally manage their diabetes to minimize complications. Likewise, when a patient is correctly diagnosed, they can be empowered to manage their diabetes with the appropriate therapies. PMID- 19302692 TI - Parental disconnect between perceived and actual weight status of children: a metasynthesis of the current research. AB - PURPOSE: Obesity is one of the most common chronic diseases in childhood. Many studies offer a variety of explanations for the alarming increase in childhood obesity; however, none discuss why an apparent disconnect exists in parental perceptions of their child's weight status. The purpose of this article was to review the current research literature on parental perceptions about their children's weight. DATA SOURCE: The articles included in this review were retrieved through a literature search using PubMed. Key words used to obtain relevant articles include childhood obesity, childhood overweight, and parental perception. CONCLUSIONS: Several studies looked at parental perceptions of childhood obesity generated from the United Kingdom, Australia, Italy, and the United States. Universally, parents were more likely to misperceive their child's weight. This was especially true for parents who were themselves overweight. IMPLICATIONS FOR PRACTICE: If parents do not recognize their child as at risk for overweight or overweight, they cannot intervene to diminish the risk factors for pediatric obesity and its related complications. More research is needed to identify why this phenomenon occurs. Only then can effective interventions be initiated. PMID- 19302693 TI - Nurse practitioner-led multidisciplinary teams to improve chronic illness care: the unique strengths of nurse practitioners applied to shared medical appointments/group visits. AB - PURPOSE: To describe the roles of nurse practitioners (NPs) in a novel model of healthcare delivery for patients with chronic disease: shared medical appointments (SMAs)/group visits based on the chronic care model (CCM). To map the specific skills of NPs to the six elements of the CCM: self-management, decision support, delivery system design, clinical information systems, community resources, and organizational support. DATA SOURCES: Case studies of three disease-specific multidisciplinary SMAs (diabetes, heart failure, and hypertension) in which NPs played a leadership role. CONCLUSIONS: NPs have multiple roles in development, implementation, and sustainability of SMAs as quality improvement interventions. Although the specific skills of NPs map out all six elements of the CCM, in our context, they had the greatest role in self management, decision support, and delivery system design. IMPLICATIONS FOR PRACTICE: With the increasing numbers of patients with chronic illnesses, healthcare systems are increasingly challenged to provide necessary care and empower patients to participate in that care. NPs can play a key role in helping to meet these challenges. PMID- 19302694 TI - The prevalence of impaired glucose metabolism in Hispanics with two or more risk factors for metabolic syndrome in the primary care setting. AB - PURPOSE: The purposes of this observational prospective study were (a) to identify the prevalence of undiagnosed impaired glucose metabolism (IGM) including impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and type 2 diabetes mellitus (T2DM) in 55 Hispanic subjects with two or more risk factors for the metabolic syndrome, (b) to examine the association between glucose metabolism and cardiometabolic risk factors (CMRF), including metabolic syndrome components, and (c) to identify predictors of IGM. DATA SOURCES: Subjects underwent a physical examination and a 2-h 75-g oral glucose tolerance test. Data were analyzed using SAS v9.1 with p < or = .05 considered significant. Nonparametric tests were applied including Mann-Whitney-Wilcoxon test and Spearman correlation coefficient. Stepwise logistic multiple regression was used to predict IGM. CONCLUSIONS: Twenty-five patients (46%) had IGM (18% IFG, 15% IGT, and 13%T2DM). Normal fasting glucose was found in 48% of subjects who had IGM. Lipid abnormalities were present in 98% including elevated triglycerides (TG 66%), total cholesterol (48%), low-density lipoprotein (68.8%), and low high density lipoprotein (67.9%). Twenty-nine percent had body mass index (BMI) >25 kg/m(2) and 62% had BMI >30 kg/m, hypertension (24%), and elevated high sensitivity C-reactive protein (63%), and mean number of cardiometabolic risk factors (#CMRF) was 4.5. Mean values for each risk factor were no different between groups except for #CMRF (p = .0001) and TG (p = .0001). Total #CMRF was the best predictor of IGM. IMPLICATIONS FOR PRACTICE: The prevalence of IGM is extremely high in Hispanics with metabolic syndrome. Screening for IGM with fasting blood glucose alone underestimates the prevalence of IGM in this population. In subjects with multiple CMRF, screening at lower levels of BMI is warranted. PMID- 19302695 TI - The advance directive prevalence in long-term care: a comparison of relationships between a nurse practitioner healthcare model and a traditional healthcare model. AB - PURPOSE: The purpose of this study was to examine rates of completion of advance directives (ADs) among institutionalized older adults in three geographically diverse areas of the country--Arizona, Georgia, and Massachusetts. Comparisons among four variables--gender, race, education, and type of healthcare model (Evercare vs. non-Evercare), related to AD completion rates were examined. DATA SOURCES: This study was a secondary data analysis using deidentified data from 11,775 older adults enrolled in the Evercare healthcare model to 91,443 non Evercare older adults (Minimum Data Set) during the last quarter of 2004. Chi square analysis was used to examine any differences in gender, race, education, and healthcare model associated with the completion rates of ADs. CONCLUSIONS: The Evercare healthcare model that used nurse practitioners (NPs) consistently had significantly higher (p < .001) completion rates of ADs compared to the non Evercare healthcare model that did not use NPs. Black people and white people in the Evercare healthcare model had similar rates of AD completion (p > .001), which is contrary to previous findings where black people had a lower completion rate. Males and females in the Evercare healthcare model had similar rates of AD completion (p > .001), which is also contrary to previous findings where females had a higher completion rate. Finally, older adults with a high school education or less and older adults with greater than a high school education in the Evercare healthcare model had similar rates of AD completion (p > .001), which is contrary to previous findings where individuals with increased education had a higher completion rate. IMPLICATIONS FOR PRACTICE: With the increasing number of older adults in the general and the long-term care population, older adults should be encouraged to complete their ADs when discussing their medical decisions with their healthcare providers. Through the use of the Evercare healthcare model, NPs are well prepared to assist their clients and families in identifying these decisions. As a result, a significantly greater proportion of ADs have been completed by individuals enrolled in the Evercare healthcare model when compared to non-Evercare individuals living in long-term care settings. By using this model, Evercare NPs ensure that the specific medical choices of their patients are carried out. PMID- 19302696 TI - 2008 AANP National NP Compensation Survey. PMID- 19302697 TI - Sexual function among women with stress incontinence after using transobturator vaginal tape, and its correlation with patient's expectations. AB - OBJECTIVE: To evaluate changes in female sexual function after a transobturator vaginal tape (TOT) procedure for treating genuine stress urinary incontinence (SUI), and its correlation with patient's expectation. PATIENTS AND METHODS: The study included women treated with a suburethral TOT for genuine SUI, neurologically intact, heterosexual and married, aged >18 years, with no previous history of malignancy, pelvic radiotherapy and no other associated surgical or psychological diseases. Patients were interviewed before surgery and with the aid of a questionnaire including female sexual function, the Beck depression indices and their expectation of sexual function after surgery. RESULTS: Sixty-two premenopausal sexually active women were included (mean age 40.5 years). The cure rate from SUI was 92%, 89%, 87% and 84% at 6, 12, 18 and 24 months, respectively. All patients attended the visit before and the first visit after surgery, while 71%, 42% and 24% were assessed at the 12-, 18 and 24-month visits, respectively. The mean follow-up was 12 months. Fifty-two women resumed their sexual activity early within the 8 weeks after surgery and the frequency of coitus in more than two-thirds of patients was at least once per month. The number of women who expected either looseness or tightness of the vagina was more than that estimated from patient perceptions. There was a difference between the patient's sensation of vaginal length abnormalities during coitus (two women) and patient expectation (18 women). CONCLUSION: Although the TOT is effective for treating SUI, counselling the patient and her partner is important in correcting false ideas and expectations about future sexual activity. Indeed, sexual dysfunction is reported after vaginal surgery, with a physiological and psychological background. Further assessment should be used to characterize sexual dysfunction after vaginal surgery for SUI to find new solutions. PMID- 19302698 TI - Development and evaluation of a multiplex test for the detection of atypical bacterial DNA in community-acquired pneumonia during childhood. AB - An incorrect or late diagnosis can lead to an increase in the morbidity and mortality caused by pneumonia, and the availability of a rapid and accurate microbiological test to verify the aetiology is imperative. This study evaluated a molecular test for the identification of the bacterial cause of atypical community-acquired pneumonia (ACAP). Fifty-four children with pneumonia were studied using bacteriological cultures, Mycoplasma pneumoniae, Coxiella burnetii, Chlamydophila pneumoniae and Legionella spp. serology, and Streptococcus pneumoniae and Legionella antigens. Simultaneously, the presence of bacterial and fungal DNA was tested for in respiratory secretion samples using the Vircell SL kit, including multiplex PCR and amplicon detection by means of line blots. There were 14 cases of ACAP caused by M. pneumoniae, with positive kit results for 13 of them, and two cases of Q-fever, with negative kit results for Coxiella burnetii. The test was negative in the remaining 38 cases (one staphylococcal pneumonia, 20 Streptococcus pneumoniae pneumonias, and 17 probable viral pneumonias). The sensitivity of the test for the detection of M. pneumoniae was 92.8% and the specificity was 100%. The Vircell SL kit allows detection of M. pneumoniae DNA in respiratory secretion samples from children with ACAP. PMID- 19302699 TI - Calculation of NNTs in RCTs with time-to-event outcomes: a literature review. AB - BACKGROUND: The number needed to treat (NNT) is a well-known effect measure for reporting the results of clinical trials. In the case of time-to-event outcomes, the calculation of NNTs is more difficult than in the case of binary data. The frequency of using NNTs to report results of randomised controlled trials (RCT) investigating time-to-event outcomes and the adequacy of the applied calculation methods are unknown. METHODS: We searched in PubMed for RCTs with parallel group design and individual randomisation, published in four frequently cited journals between 2003 and 2005. We evaluated the type of outcome, the frequency of reporting NNTs with corresponding confidence intervals, and assessed the adequacy of the methods used to calculate NNTs in the case of time-to-event outcomes. RESULTS: The search resulted in 734 eligible RCTs. Of these, 373 RCTs investigated time-to-event outcomes and 361 analyzed binary data. In total, 62 articles reported NNTs (34 articles with time-to-event outcomes, 28 articles with binary outcomes). Of the 34 articles reporting NNTs derived from time-to-event outcomes, only 17 applied an appropriate calculation method. Of the 62 articles reporting NNTs, only 21 articles presented corresponding confidence intervals. CONCLUSION: The NNT is used as effect measure to present the results from RCTs with binary and time-to-event outcomes in the current medical literature. In the case of time-to-event data incorrect methods were frequently applied. Confidence intervals for NNTs were given in one third of the NNT reporting articles only. In summary, there is much room for improvement in the application of NNTs to present results of RCTs, especially where the outcome is time to an event. PMID- 19302700 TI - Using an expiratory resistor, arterial pulse pressure variations predict fluid responsiveness during spontaneous breathing: an experimental porcine study. AB - INTRODUCTION: Fluid responsiveness prediction is difficult in spontaneously breathing patients. Because the swings in intrathoracic pressure are minor during spontaneous breathing, dynamic parameters like pulse pressure variation (PPV) and systolic pressure variation (SPV) are usually small. We hypothesized that during spontaneous breathing, inspiratory and/or expiratory resistors could induce high arterial pressure variations at hypovolemia and low variations at normovolemia and hypervolemia. Furthermore, we hypothesized that SPV and PPV could predict fluid responsiveness under these conditions. METHODS: Eight prone, anesthetized and spontaneously breathing pigs (20 to 25 kg) were subjected to a sequence of 30% hypovolemia, normovolemia, and 20% and 40% hypervolemia. At each volemic level, the pigs breathed in a randomized order either through an inspiratory and/or an expiratory threshold resistor (7.5 cmH2O) or only through the tracheal tube without any resistor. Hemodynamic and respiratory variables were measured during the breathing modes. Fluid responsiveness was defined as a 15% increase in stroke volume (DeltaSV) following fluid loading. RESULTS: Stroke volume was significantly lower at hypovolemia compared with normovolemia, but no differences were found between normovolemia and 20% or 40% hypervolemia. Compared with breathing through no resistor, SPV was magnified by all resistors at hypovolemia whereas there were no changes at normovolemia and hypervolemia. PPV was magnified by the inspiratory resistor and the combined inspiratory and expiratory resistor. Regression analysis of SPV or PPV versus DeltaSV showed the highest R2 (0.83 for SPV and 0.52 for PPV) when the expiratory resistor was applied. The corresponding sensitivity and specificity for prediction of fluid responsiveness were 100% and 100%, respectively, for SPV and 100% and 81%, respectively, for PPV. CONCLUSIONS: Inspiratory and/or expiratory threshold resistors magnified SPV and PPV in spontaneously breathing pigs during hypovolemia. Using the expiratory resistor SPV and PPV predicted fluid responsiveness with good sensitivity and specificity. PMID- 19302701 TI - MarVis: a tool for clustering and visualization of metabolic biomarkers. AB - BACKGROUND: A central goal of experimental studies in systems biology is to identify meaningful markers that are hidden within a diffuse background of data originating from large-scale analytical intensity measurements as obtained from metabolomic experiments. Intensity-based clustering is an unsupervised approach to the identification of metabolic markers based on the grouping of similar intensity profiles. A major problem of this basic approach is that in general there is no prior information about an adequate number of biologically relevant clusters. RESULTS: We present the tool MarVis (Marker Visualization) for data mining on intensity-based profiles using one-dimensional self-organizing maps (1D SOMs). MarVis can import and export customizable CSV (Comma Separated Values) files and provides aggregation and normalization routines for preprocessing of intensity profiles that contain repeated measurements for a number of different experimental conditions. Robust clustering is then achieved by training of an 1D SOM model, which introduces a similarity-based ordering of the intensity profiles. The ordering allows a convenient visualization of the intensity variations within the data and facilitates an interactive aggregation of clusters into larger blocks. The intensity-based visualization is combined with the presentation of additional data attributes, which can further support the analysis of experimental data. CONCLUSION: MarVis is a user-friendly and interactive tool for exploration of complex pattern variation in a large set of experimental intensity profiles. The application of 1D-SOMs gives a convenient overview on relevant profiles and groups of profiles. The specialized visualization effectively supports researchers in analyzing a large number of putative clusters, even though the true number of biologically meaningful groups is unknown. Although MarVis has been developed for the analysis of metabolomic data, the tool may be applied to gene expression data as well. PMID- 19302702 TI - Identification of subpopulations in mesenchymal stem cell-like cultures from human umbilical cord. AB - BACKGROUND: A variety of cell types can be identified in the adherent fraction of bone marrow mononuclear cells including more primitive and embryonic-like stem cells, mesenchymal stem cells (MSC), lineage-committed progenitors as well as mature cells such as osteoblasts and fibroblasts. Different methods are described for the isolation of single bone marrow stem cell subpopulations - beginning from ordinary size sieving, long term cultivation under specific conditions to FACS based approaches. Besides bone marrow-derived subpopulations, also other tissues including human umbilical cord (UC) have been recently suggested to provide a potential source for MSC. Although of clinical importance, these UC-derived MSC populations remain to be characterized. It was thus the aim of the present study to identify possible subpopulations in cultures of MSC-like cells obtained from UC. We used counterflow centrifugal elutriation (CCE) as a novel strategy to successfully address this question. RESULTS: UC-derived primary cells were separated by CCE and revealed differentially-sized populations in the fractions. Thus, a subpopulation with an average diameter of about 11 mum and a small flat cell body was compared to a large sized subpopulation of about 19 mum average diameter. Flow cytometric analysis revealed the expression of certain MSC stem cell markers including CD44, CD73, CD90 and CD105, respectively, although these markers were expressed at higher levels in the small-sized population. Moreover, this small-sized subpopulation exhibited a higher proliferative capacity as compared to the total UC-derived primary cultures and the large-sized cells and demonstrated a reduced amount of aging cells. CONCLUSION: Using the CCE technique, we were the first to demonstrate a subpopulation of small-sized UC derived primary cells carrying MSC-like characteristics according to the presence of various mesenchymal stem cell markers. This is also supported by the high proliferative capacity of these MSC-like cells as compared to whole primary culture or other UC-derived subpopulations. The accumulation of a self-renewing MSC-like subpopulation by CCE with low expression levels of the aging marker senescence-associated beta-galactosidase provides a valuable tool in the regenerative medicine and an alternative to bone-marrow-derived MSC. PMID- 19302703 TI - Hypoxia inducible factor-1alpha correlates with vascular endothelial growth factor A and C indicating worse prognosis in clear cell renal cell carcinoma. AB - BACKGROUND: The role of angiogenesis in the pathogenesis of renal cell carcinoma is well recognized, however, the influence of tumor cells in this activity has not yet been fully clarified. The aim of this study was to analyze the expression of hypoxia inducible factor-1alpha (HIF-1alpha), a regulatory factor of angiogenic switch, in comparison to vascular endothelial growth factor A and C (VEGF-A and VEGF-C), recognized to be involved in blood and lymph vessel neoangiogenesis, with potential association in the prognosis of patients with renal cell carcinoma. METHODS: Ninety-four patients with diagnosis of clear cell renal cell carcinomas (CCRCC), all clinicopathological characteristics and overall survival were unrolled in this study. Immunohistochemicaly VEGF-A, VEGF C, HIF-1alpha and Ki67 were detected on tumor cells and the staining was performed on tissue microarrays (TMA). The staining was evaluated as a percentage of cytoplasmic or nuclear positive tumor cells. RESULTS: Variable expression of all three proteins was confirmed. Both angiogenic factors demonstrated perimembranous or diffuse cytoplasmic staining, with diffuse pattern positively associated (p < 0.001). Nuclear HIF-1alpha expression (nHIF-1alpha) showed inverse correlation with diffuse cytoplasmic VEGF-A (p = 0.002) and VEGF-C (p = 0.053), while cytoplasmic HIF-1alpha expression (cHIF-1alpha) showed positive correlation with diffuse staining of both angiogenic factors (p < 0.001; p < 0.001, respectively). In comparison to clinicopathological characteristics, a higher nuclear grade (p = 0.006; p < 0.001, respectively), larger tumor size (p = 0.009; p = 0.015, respectively), higher stage (p = 0.023; p = 0.027, respectively) and shorter survival (p = 0.018; p = 0.024, respectively) were associated with overexpression of cHIF-1alpha and diffuse cytoplasmic VEGF-A expression. In contrary, overexpression of nHIF-1alpha was associated with better diagnostic parameters i.e. lower nuclear grade (p = 0.006), smaller tumor size (p = 0.057), and longer survival (p = 0.005). CONCLUSION: Overexpression of VEGF-A and cHIF-1alpha in tumor cells highlights a more aggressive subtype of CCRCC that might have some clinical implications. The significance of nHIF-1alpha expression associated with better differentiated tumors should be further elucidated. PMID- 19302704 TI - Mutations in genes involved in nonsense mediated decay ameliorate the phenotype of sel-12 mutants with amber stop mutations in Caenorhabditis elegans. AB - BACKGROUND: Presenilin proteins are part of a complex of proteins that can cleave many type I transmembrane proteins, including Notch Receptors and the Amyloid Precursor Protein, in the middle of the transmembrane domain. Dominant mutations in the human presenilin genes PS1 and PS2 lead to Familial Alzheimer's disease. Mutations in the Caenorhabditis elegans sel-12 presenilin gene cause a highly penetrant egg-laying defect due to reduction of signalling through the lin 12/Notch receptor. Mutations in six spr genes (for suppressor of presenilin) are known to strongly suppress sel-12. Mutations in most strong spr genes suppress sel-12 by de-repressing the transcription of the largely functionally equivalent hop-1 presenilin gene. However, how mutations in the spr-2 gene suppress sel-12 is unknown. RESULTS: We show that spr-2 mutations increase the levels of sel-12 transcripts with Premature translation Termination Codons (PTCs) in embryos and L1 larvae. mRNA transcripts from sel-12 alleles with PTCs undergo degradation by a process known as Nonsense Mediated Decay (NMD). However, spr-2 mutations do not appear to affect NMD. Mutations in the smg genes, which are required for NMD, can restore sel-12(PTC) transcript levels and ameliorate the phenotype of sel-12 mutants with amber PTCs. However, the phenotypic suppression of sel-12 by smg genes is nowhere near as strong as the effect of previously characterized spr mutations including spr-2. Consistent with this, we have identified only two mutations in smg genes among the more than 100 spr mutations recovered in genetic screens. CONCLUSION: spr-2 mutations do not suppress sel-12 by affecting NMD of sel-12(PTC) transcripts and appear to have a novel mechanism of suppression. The fact that mutations in smg genes can ameliorate the phenotype of sel-12 alleles with amber PTCs suggests that some read-through of sel-12(amber) alleles occurs in smg backgrounds. PMID- 19302705 TI - Testing for the association of the KIAA1109/Tenr/IL2/IL21 gene region with rheumatoid arthritis in a European family-based study. AB - INTRODUCTION: A candidate gene approach, in a large case-control association study in the Dutch population, has shown that a 480 kb block on chromosome 4q27 encompassing KIAA1109/Tenr/IL2/IL21 genes is associated with rheumatoid arthritis. Compared with case-control association studies, family-based studies have the added advantage of controlling potential differences in population structure. Therefore, our aim was to test this association in populations of European origin by using a family-based approach. METHODS: A total of 1,302 West European white individuals from 434 trio families were genotyped for the rs4505848, rs11732095, rs6822844, rs4492018 and rs1398553 polymorphisms using the TaqMan Allelic discrimination assay (Applied Biosystems). The genetic association analyses for each SNP and haplotype were performed using the Transmission Disequilibrium Test and the genotype relative risk. RESULTS: We observed evidence for association of the heterozygous rs4505848-AG genotype with rheumatoid arthritis (P = 0.04); however, no significance was found after Bonferroni correction. In concordance with previous findings in the Dutch population, we observed a trend of undertransmission for the rs6822844-T allele and rs6822844-GT genotype to rheumatoid arthritis patients. We further investigated the five SNP haplotypes of the KIAA1109/Tenr/IL2/IL21 gene region. We observed, as described in the Dutch population, a nonsignificant undertransmission of the AATGG haplotype to rheumatoid arthritis patients. CONCLUSIONS: Using a family-based study, we have provided a trend for the association of the KIAA1109/Tenr/IL2/IL21 gene region with rheumatoid arthritis in populations of European descent. Nevertheless, we failed to replicate a significant association of this region in our rheumatoid arthritis family sample. Further investigation of this region, including detection and testing of all variants, is required to confirm rheumatoid arthritis association. PMID- 19302706 TI - Conflict among Iranian hospital nurses: a qualitative study. AB - BACKGROUND: This study aims to explore the experience of conflict as perceived by Iranian hospital nurses in Tehran, Islamic Republic of Iran. Although conflict control approaches have been extensively researched throughout the world, no research-based data are available on the perception of conflict and effective resolutions among hospital nurses in Iran. METHODS: A qualitative research approach was used to explore how Iranian hospital nurses perceive and resolve conflicts at work. A purposive sample of 30 hospital nurses and nurse managers was selected to obtain data by means of in-depth semi structured interviews. Data were analysed by means of the content analysis method. RESULTS: The emerging themes were: (1) the nurses' perceptions and reactions to conflict; (2) organizational structure; (3) hospital management style; (4) the nature and conditions of job assignment; (5) individual characteristics; (6) mutual understanding and interaction; and (7) the consequences of conflict. The first six themes describe the sources of the conflict as well as strategies to manage them. CONCLUSION: How nurses perceive conflict influences how they react to it. Sources of conflict are embedded in the characteristics of nurses and the nursing system, but at the same time these characteristics can be seen as strategies to resolve conflict. We found mutual understanding and interaction to be the main factor able to prevent and resolve conflict effectively. We therefore recommend that nurses and nurse managers encourage any virtues and activities that increase such understanding and interaction. Finally, as conflict can destroy individual nurses as well as the nursing system, we must act to control it effectively. PMID- 19302707 TI - Interferon beta-1a for the maintenance of remission in patients with Crohn's disease: results of a phase II dose-finding study. AB - BACKGROUND: Crohn's disease (CD) and multiple sclerosis (MS) share common pathogenic processes. Interferon (IFN) beta-1a is effective and generally well tolerated in patients with MS and has been shown to down-regulate the expression of interleukin-12, a cytokine that is thought to be involved in mucosal degeneration in CD. IFN beta-1a therefore offers promise as a treatment for CD. METHODS: In this multicentre, double-blind, placebo-controlled, phase II, dose finding study, patients with steroid-induced clinical remissions of CD were randomized 1:1:1:1 to subcutaneous IFN beta-1a: 66 mcg three times weekly (tiw), 44 mcg tiw, 44 mcg twice weekly (biw), or matching placebo tiw with steroid tapering. The primary endpoint was the proportion of patients relapse-free at Week 26. Safety was also assessed. RESULTS: This study was terminated early following a planned interim analysis at 26 weeks. Of the planned 192 patients, 67 were randomized to treatment: placebo (n = 16), or IFN beta-1a 44 mcg biw (n = 17), 44 mcg tiw (n = 16) or 66 mcg tiw (n = 18). In total, 20/67 patients (29.9%) completed 26 weeks and 7 patients (10.4%) completed 52 weeks. The proportion of patients who remained relapse-free at Week 26 did not differ significantly between the placebo group (5/16, 31%) and the IFN beta-1a 44 mcg biw (6/17, 35%; p = 0.497), 44 mcg tiw (7/16, 44%; p = 0.280) or 66 mcg tiw (2/18, 11%; p = 0.333) groups. There was little difference between treatment groups in secondary efficacy endpoints. IFN beta-1a was generally well tolerated at all doses. Adverse events (AEs) were generally mild or moderate in IFN beta-1a-treated patients, with the most common AEs (influenza-like symptoms, headache, injection site reactions) being similar to those reported with IFN beta-1a in MS. CONCLUSION: There was no difference in efficacy between patients with CD receiving IFN beta-1a or placebo. However, these results should be considered in the context of the low patient numbers and high dropout rate. Overall, IFN beta 1a was generally well tolerated, with a safety profile that was consistent with previous experience in MS. TRIAL REGISTRATION: ClinicalTrials.gov NCT00304252. PMID- 19302708 TI - Transcriptional signatures of BALB/c mouse macrophages housing multiplying Leishmania amazonensis amastigotes. AB - BACKGROUND: Mammal macrophages (MPhi) display a wide range of functions which contribute to surveying and maintaining tissue integrity. One such function is phagocytosis, a process known to be subverted by parasites like Leishmania (L). Indeed, the intracellular development of L. amazonensis amastigote relies on the biogenesis and dynamic remodelling of a phagolysosome, termed the parasitophorous vacuole, primarily within dermal MPhi. RESULTS: Using BALB/c mouse bone marrow derived MPhi loaded or not with amastigotes, we analyzed the transcriptional signatures of MPhi 24 h later, when the amastigote population was growing. Total RNA from MPhi cultures were processed and hybridized onto Affymetrix Mouse430_2 GeneChips, and some transcripts were also analyzed by Real-Time quantitative PCR (RTQPCR). A total of 1,248 probe-sets showed significant differential expression. Comparable fold-change values were obtained between the Affymetrix technology and the RTQPCR method. Ingenuity Pathway Analysis software pinpointed the up regulation of the sterol biosynthesis pathway (p-value = 1.31e-02) involving several genes (1.95 to 4.30 fold change values), and the modulation of various genes involved in polyamine synthesis and in pro/counter-inflammatory signalling. CONCLUSION: Our findings suggest that the amastigote growth relies on early coordinated gene expression of the MPhi lipid and polyamine pathways. Moreover, these MPhi hosting multiplying L. amazonensis amastigotes display a transcriptional profile biased towards parasite-and host tissue-protective processes. PMID- 19302709 TI - New scoring schema for finding motifs in DNA Sequences. AB - BACKGROUND: Pattern discovery in DNA sequences is one of the most fundamental problems in molecular biology with important applications in finding regulatory signals and transcription factor binding sites. An important task in this problem is to search (or predict) known binding sites in a new DNA sequence. For this reason, all subsequences of the given DNA sequence are scored based on an scoring function and the prediction is done by selecting the best score. By assuming no dependency between binding site base positions, most of the available tools for known binding site prediction are designed. Recently Tomovic and Oakeley investigated the statistical basis for either a claim of dependence or independence, to determine whether such a claim is generally true, and they presented a scoring function for binding site prediction based on the dependency between binding site base positions. Our primary objective is to investigate the scoring functions which can be used in known binding site prediction based on the assumption of dependency or independency in binding site base positions. RESULTS: We propose a new scoring function based on the dependency between all positions in biding site base positions. This scoring function uses joint information content and mutual information as a measure of dependency between positions in transcription factor binding site. Our method for modeling dependencies is simply an extension of position independency methods. We evaluate our new scoring function on the real data sets extracted from JASPAR and TRANSFAC data bases, and compare the obtained results with two other well known scoring functions. CONCLUSION: The results demonstrate that the new approach improves known binding site discovery and show that the joint information content and mutual information provide a better and more general criterion to investigate the relationships between positions in the TFBS. Our scoring function is formulated by simple mathematical calculations. By implementing our method on several biological data sets, it can be induced that this method performs better than methods that do not consider dependencies. PMID- 19302710 TI - Bacteriological assessment of urban water sources in Khamis Mushait Governorate, southwestern Saudi Arabia. AB - BACKGROUND: Urban water sources of Khamis Mushait Governorate, southwestern Saudi Arabia, were studied to assess their bacteriological characteristics and suitability for potable purposes. A cross-sectional epidemiological method was adopted to investigate the four main urban water sources (i.e. bottled, desalinated, surface, and well water). These were sampled and examined between February and June 2007. RESULTS: A total of 95 water samples from bottled, desalinated, surface, and well water were collected randomly from the study area using different gathering and analysing techniques. The bacteriological examination of water samples included the most probable number of presumptive coliforms, faecal coliforms, and faecal streptococci (MPN/100 ml). The results showed that the total coliform count (MPN/100 ml) was not detected in any samples taken from bottled water, while it was detected in those taken from desalinated, surface, and well water: percentages were 12.9, 80.0, and 100.0, respectively. Faecal coliforms were detected in desalinated, surface, and well water, with percentages of 3.23, 60.0 and 87.88, respectively. About 6.45% of desalinated water, 53.33% of surface water, and 57.58% of well water was found positive for faecal streptococci. Colonies of coliforms were identified in different micro organisms with various percentages. CONCLUSION: Water derived from traditional sources (wells) showed increases in most of the investigated bacteriological parameters, followed by surface water as compared to bottled or desalinated water. This may be attributed to the fact that well and surface water are at risk of contamination as indicated by the higher levels of most bacteriological parameters. Moreover, well water is exposed to point sources of pollution such as septic wells and domestic and farming effluents, as well as to soil with a high humus content. The lower bacteriological characteristics in samples from bottled water indicate that it is satisfactory for human drinking purposes. Contamination of desalinated water that is the main urban water source may occur during transportation from the desalination plant or in the house reservoir of the consumer. Improving and expanding the existing water treatment and sanitation systems is more likely to provide safe and sustainable sources of water over the long term. Strict hygienic measures should be applied to improve water quality and to avoid deleterious effects on public health, by using periodical monitoring programmes to detect sewage pollution running over local hydrological networks and valleys. PMID- 19302711 TI - Overexpression of UbcH10 alternates the cell cycle profile and accelerate the tumor proliferation in colon cancer. AB - BACKGROUND: UbcH10 participates in proper metaphase to anaphase transition, and abrogation of UbcH10 results in the premature separation of sister chromatids. To assess the potential role of UbcH10 in colon cancer progression, we analyzed the clinicopathological relevance of UbcH10 in colon cancer. METHODS: We firstly screened the expression profile of UbcH10 in various types of cancer tissues as well as cell lines. Thereafter, using the colon cancer cells line, we manipulated the expression of UbcH10 and evaluated the cell cycle profile and cellular proliferations. Furthermore, the clinicopathological significance of UbcH10 was immunohistologically evaluated in patients with colon cancer. Statistical analysis was performed using the student's t-test and Chi-square test. RESULTS: Using the colon cancer cells, depletion of UbcH10 resulted in suppression of cellular growth whereas overexpression of UbcH10 promoted the cellular growth and oncogenic cellular growth. Mitotic population was markedly alternated by the manipulation of UbcH10 expression. Immunohistochemical analysis indicated that UbcH10 was significantly higher in colon cancer tissue compared with normal colon epithelia. Furthermore, the clinicopathological evaluation revealed that UbcH10 was associated with high-grade histological tumors. CONCLUSION: The results show the clinicopathological significance of UbcH10 in the progression of colon cancer. Thus UbcH10 may act as a novel biomarker in patients with colon cancer. PMID- 19302712 TI - Sugarcane genes associated with sucrose content. AB - BACKGROUND: Sucrose content is a highly desirable trait in sugarcane as the worldwide demand for cost-effective biofuels surges. Sugarcane cultivars differ in their capacity to accumulate sucrose and breeding programs routinely perform crosses to identify genotypes able to produce more sucrose. Sucrose content in the mature internodes reach around 20% of the culms dry weight. Genotypes in the populations reflect their genetic program and may display contrasting growth, development, and physiology, all of which affect carbohydrate metabolism. Few studies have profiled gene expression related to sugarcane's sugar content. The identification of signal transduction components and transcription factors that might regulate sugar accumulation is highly desirable if we are to improve this characteristic of sugarcane plants. RESULTS: We have evaluated thirty genotypes that have different Brix (sugar) levels and identified genes differentially expressed in internodes using cDNA microarrays. These genes were compared to existing gene expression data for sugarcane plants subjected to diverse stress and hormone treatments. The comparisons revealed a strong overlap between the drought and sucrose-content datasets and a limited overlap with ABA signaling. Genes associated with sucrose content were extensively validated by qRT-PCR, which highlighted several protein kinases and transcription factors that are likely to be regulators of sucrose accumulation. The data also indicate that aquaporins, as well as lignin biosynthesis and cell wall metabolism genes, are strongly related to sucrose accumulation. Moreover, sucrose-associated genes were shown to be directly responsive to short term sucrose stimuli, confirming their role in sugar-related pathways. CONCLUSION: Gene expression analysis of sugarcane populations contrasting for sucrose content indicated a possible overlap with drought and cell wall metabolism processes and suggested signaling and transcriptional regulators to be used as molecular markers in breeding programs. Transgenic research is necessary to further clarify the role of the genes and define targets useful for sugarcane improvement programs based on transgenic plants. PMID- 19302713 TI - Crystalloids versus colloids for goal-directed fluid therapy in major surgery. AB - INTRODUCTION: Perioperative hypovolemia arises frequently and contributes to intestinal hypoperfusion and subsequent postoperative complications. Goal directed fluid therapy might reduce these complications. The aim of this study was to compare the effects of goal-directed administration of crystalloids and colloids on the distribution of systemic, hepatosplanchnic, and microcirculatory (small intestine) blood flow after major abdominal surgery in a clinically relevant pig model. METHODS: Twenty-seven pigs were anesthetized and mechanically ventilated and underwent open laparotomy. They were randomly assigned to one of three treatment groups: the restricted Ringer lactate (R-RL) group (n = 9) received 3 mL/kg per hour of RL, the goal-directed RL (GD-RL) group (n = 9) received 3 mL/kg per hour of RL and intermittent boluses of 250 mL of RL, and the goal-directed colloid (GD-C) group (n = 9) received 3 mL/kg per hour of RL and boluses of 250 mL of 6% hydroxyethyl starch (130/0.4). The latter two groups received a bolus infusion when mixed venous oxygen saturation was below 60% ('lockout' time of 30 minutes). Regional blood flow was measured in the superior mesenteric artery and the celiac trunk. In the small bowel, microcirculatory blood flow was measured using laser Doppler flowmetry. Intestinal tissue oxygen tension was measured with intramural Clark-type electrodes. RESULTS: After 4 hours of treatment, arterial blood pressure, cardiac output, mesenteric artery flow, and mixed oxygen saturation were significantly higher in the GD-C and GD-RL groups than in the R-RL group. Microcirculatory flow in the intestinal mucosa increased by 50% in the GD-C group but remained unchanged in the other two groups. Likewise, tissue oxygen tension in the intestine increased by 30% in the GD-C group but remained unchanged in the GD-RL group and decreased by 18% in the R-RL group. Mesenteric venous glucose concentrations were higher and lactate levels were lower in the GD-C group compared with the two crystalloid groups. CONCLUSIONS: Goal-directed colloid administration markedly increased microcirculatory blood flow in the small intestine and intestinal tissue oxygen tension after abdominal surgery. In contrast, goal-directed crystalloid and restricted crystalloid administrations had no such effects. Additionally, mesenteric venous glucose and lactate concentrations suggest that intestinal cellular substrate levels were higher in the colloid-treated than in the crystalloid-treated animals. These results support the notion that perioperative goal-directed therapy with colloids might be beneficial during major abdominal surgery. PMID- 19302714 TI - Editorial on hypothesis and objectives in clinical trials: superiority, equivalence and non-inferiority. PMID- 19302715 TI - Statistical and visual differentiation of subcellular imaging. AB - BACKGROUND: Automated microscopy technologies have led to a rapid growth in imaging data on a scale comparable to that of the genomic revolution. High throughput screens are now being performed to determine the localisation of all of proteins in a proteome. Closer to the bench, large image sets of proteins in treated and untreated cells are being captured on a daily basis to determine function and interactions. Hence there is a need for new methodologies and protocols to test for difference in subcellular imaging both to remove bias and enable throughput. Here we introduce a novel method of statistical testing, and supporting software, to give a rigorous test for difference in imaging. We also outline the key questions and steps in establishing an analysis pipeline. RESULTS: The methodology is tested on a high throughput set of images of 10 subcellular localisations, and it is shown that the localisations may be distinguished to a statistically significant degree with as few as 12 images of each. Further, subtle changes in a protein's distribution between nocodazole treated and control experiments are shown to be detectable. The effect of outlier images is also examined and it is shown that while the significance of the test may be reduced by outliers this may be compensated for by utilising more images. Finally, the test is compared to previous work and shown to be more sensitive in detecting difference. The methodology has been implemented within the iCluster system for visualising and clustering bio-image sets. CONCLUSION: The aim here is to establish a methodology and protocol for testing for difference in subcellular imaging, and to provide tools to do so. While iCluster is applicable to moderate (<1000) size image sets, the statistical test is simple to implement and will readily be adapted to high throughput pipelines to provide more sensitive discrimination of difference. PMID- 19302716 TI - Circulating cell death products predict clinical outcome of colorectal cancer patients. AB - BACKGROUND: Tumor cell death generates products that can be measured in the circulation of cancer patients. CK18-Asp396 (M30 antigen) is a caspase-degraded product of cytokeratin 18 (CK18), produced by apoptotic epithelial cells, and is elevated in breast and lung cancer patients. METHODS: We determined the CK18 Asp396 and total CK18 levels in plasma of 49 colorectal cancer patients, before and after surgical resection of the tumor, by ELISA. Correlations with patient and tumor characteristics were determined by Kruskal-Wallis H and Mann-Whitney U tests. Disease-free survival was determined using Kaplan-Meier methodology with Log Rank tests, and univariate and multivariate Cox proportional hazard analysis. RESULTS: Plasma CK18-Asp396 and total CK18 levels in colorectal cancer patients were related to disease stage and tumor diameter, and were predictive of disease free survival, independent of disease-stage, with hazard ratios (HR) of patients with high levels (> median) compared to those with low levels (< or = median) of 3.58 (95% CI: 1.17-11.02) and 3.58 (95% CI: 0.97-7.71), respectively. The CK18 Asp396/CK18 ratio, which decreased with tumor progression, was also predictive of disease-free survival, with a low ratio (< or = median) associated with worse disease-free survival: HR 2.78 (95% CI: 1.06-7.19). Remarkably, the plasma CK18 Asp396 and total CK18 levels after surgical removal of the tumor were also predictive of disease-free survival, with patients with high levels having a HR of 3.78 (95% CI: 0.77-18.50) and 4.12 (95% CI: 0.84-20.34), respectively, indicating that these parameters can be used also to monitor patients after surgery. CONCLUSION: CK18-Asp396 and total CK18 levels in the circulation of colorectal cancer patients are predictive of tumor progression and prognosis and might be helpful for treatment selection and monitoring of these patients. PMID- 19302717 TI - Size-partitioning of an urban aerosol to identify particle determinants involved in the proinflammatory response induced in airway epithelial cells. AB - BACKGROUND: The contribution of air particles in human cardio-respiratory diseases has been enlightened by several epidemiological studies. However the respective involvement of coarse, fine and ultrafine particles in health effects is still unclear. The aim of the present study is to determine which size fraction from a chemically characterized background aerosol has the most important short term biological effect and to decipher the determinants of such a behaviour. RESULTS: Ambient aerosols were collected at an urban background site in Paris using four 13-stage low pressure cascade impactors running in parallel (winter and summer 2005) in order to separate four size-classes (PM0.03-0.17 (defined here as ultrafine particles), PM0.17-1 (fine), PM1-2.5(intermediate) and PM2.5-10 (coarse)). Accordingly, their chemical composition and their pro inflammatory potential on human airway epithelial cells were investigated. Considering isomass exposures (same particle concentrations for each size fractions) the pro-inflammatory response characterized by Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) release was found to decrease with aerosol size with no seasonal dependency. When cells were exposed to isovolume of particle suspensions in order to respect the particle proportions observed in ambient air, the GM-CSF release was maximal with the fine fraction. In presence of a recombinant endotoxin neutralizing protein, the GM-CSF release induced by particles is reduced for all size-fractions, with exception of the ultra-fine fraction which response is not modified. The different aerosol size-fractions were found to display important chemical differences related to the various contributing primary and secondary sources and aerosol age. The GM-CSF release was correlated to the organic component of the aerosols and especially its water soluble fraction. Finally, Cytochrome P450 1A1 activity that reflects PAH bioavailability varied as a function of the season: it was maximal for the fine fraction in winter and for the ultrafine fraction in summer. CONCLUSION: In the frame of future regulations, a particular attention should thus be paid to the ultrafine/fine (here referred to as PM1) fraction due to their overwhelming anthropogenic origin and predominance in the urban aerosol and their pro inflammatory potential. PMID- 19302718 TI - Reproducibility and relative validity of dietary glycaemic index and glycaemic load assessed by the food-frequency questionnaire used in the Dutch cohorts of the European Prospective Investigation into Cancer and Nutrition. AB - Limited information is available on the reproducibility and validity of dietary glycaemic index (GI) and glycaemic load (GL) estimated by habitual diet assessment methods such as FFQ, including the FFQ used in the Dutch cohorts of the European Prospective Investigation into Cancer and Nutrition study. To examine the reproducibility and relative validity of GI and GL, we used data from 121 Dutch men and women aged 23-72 years. They completed the FFQ three times at intervals of 6 months and twelve 24-h dietary recalls (24HDR) monthly during 1991 2. GI and GL were calculated using published values. Intra-class correlation coefficients of the three repeated FFQ were 0.78 for GI and 0.74 for GL. Pearson correlation coefficients between the first FFQ and the weighted average of the 24HDR were 0.63 for both GI and GL. Weighted kappa values between the first FFQ and the average of the 24HDR (in quintiles) were 0.40 for GI and 0.41 for GL. Bland-Altman plots showed a proportional bias in GI (beta = 0.46), but not in GL (beta = 0.06). In conclusion, this FFQ can be used in epidemiological studies to investigate the relationship of GI and GL with disease risks, but the proportional bias should be taken into account when using this FFQ to assess the absolute GI values. PMID- 19302719 TI - Blackcurrant seed press residue increases tocopherol concentrations in serum and stool whilst biomarkers in stool and urine indicate increased oxidative stress in human subjects. AB - Berry seeds are a tocopherol-rich by-product of fruit processing without specific commercial value. In a human intervention study, the physiological impact of blackcurrant seed press residue (PR) was tested. Thirty-six women (aged 24 +/- 3 years; twenty non-smokers, sixteen smokers) consumed 250 g bread/d containing 8% PR for a period of 4 weeks (period 3). Comparatively, a control bread without PR (250 g/d) was tested (period 2) and baseline data were obtained (period 1). Blood, stool and 24 h urine were collected during a 5 d standardised diet within each period. Tocopherol and Fe intakes were calculated from food intake. In serum, tocopherol concentration and Fe parameters were determined. In urine, oxidative stress markers 8-oxo-2'-deoxyguanosine, 8-iso-PGF2alpha and inflammatory response marker 15-keto-dihydro-PGF2alpha were analysed. Stool tocopherol concentration, genotoxicity of faecal water (comet assay) and antioxidant capacity of stool (aromatic hydroxylation of salicylic acid) were determined. Fe and total tocopherol intake, total tocopherol concentrations in serum and stool, and genotoxicity of faecal water increased with PR bread consumption (P < 0.05). The antioxidant capacity of stool decreased between baseline and intervention, expressed by increased formation of 2,3- and 2,5 dihydroxybenzoic acid in vitro (P < 0.05). In smokers, 8-oxo-2'-deoxyguanosine increased with PR consumption (P < 0.05). Prostane concentrations were unaffected by PR bread consumption. In summary, the intake of bread containing blackcurrant PR for 4 weeks increased serum and stool total tocopherol concentrations. However, various biomarkers indicated increased oxidative stress, suggesting that consumption of ground berry seed may not be of advantage. PMID- 19302720 TI - Thin healthy women have a similar low bone mass to women with anorexia nervosa. AB - An association between anorexia nerviosa (AN) and low bone mass has been demonstrated. Bone loss associated with AN involves hormonal and nutritional impairments, though their exact contribution is not clearly established. We compared bone mass in AN patients with women of similar weight with no criteria for AN, and a third group of healthy, normal-weight, age-matched women. The study included forty-eight patients with AN, twenty-two healthy eumenorrhoeic women with low weight (LW group; BMI < 18.5 kg/m2) and twenty healthy women with BMI >18.5 kg/m2 (control group), all of similar age. We measured lean body mass, percentage fat mass, total bone mineral content (BMC) and bone mineral density in lumbar spine (BMD LS) and in total (tBMD). We measured anthropometric parameters, leptin and growth hormone. The control group had greater tBMD and BMD LS than the other groups, with no differences between the AN and LW groups. No differences were found in tBMD, BMD LS and total BMC between the restrictive (n 25) and binge purge type (n 23) in AN patients. In AN, minimum weight (P = 0.002) and percentage fat mass (P = 0.02) explained BMD LS variation (r2 0.48) and minimum weight (r2 0.42; P = 0.002) for tBMD in stepwise regression analyses. In the LW group, BMI explained BMD LS (r2 0.72; P = 0.01) and tBMD (r2 0.57; P = 0.04). We concluded that patients with AN had similar BMD to healthy thin women. Anthropometric parameters could contribute more significantly than oestrogen deficiency in the achievement of peak bone mass in AN patients. PMID- 19302721 TI - Flight take-off and walking behavior of insecticide-susceptible and - resistant strains of Sitophilus zeamais exposed to deltamethrin. AB - Insects have evolved a variety of physiological and behavioral responses to various toxins in natural and managed ecosystems. However, insect behavior is seldom considered in insecticide studies although insects are capable of changing their behavior in response to their sensory perception of insecticides, which may compromise insecticide efficacy. This is particularly serious for insect pests that are physiologically resistant to insecticides since insecticide avoidance may further compromise their management. Locomotion plays a major role determining insecticide exposure and was, therefore, considered in investigating the behavioral responses of male and female adult insects from an insecticide susceptible and two insecticide-resistant strains of the maize weevil Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), a major pest of stored cereals. Different dose-dependent behavioral responses were expected among strains with behavioral resistance less likely to occur in physiologically resistant insects since they are able to withstand higher doses of insecticide. The behavioral responses to deltamethrin-sprayed surfaces differed among the maize weevil strains. Such responses were concentration-independent for all of the strains. Stimulus-independent behavioral resistance was unrelated to physiological resistance with one resistant strain exhibiting higher rates of flight take-off and the other resistant strain exhibiting lower flight take-off. Female mobility was similar for all strains, unlike male mobility. Males of each strain exhibited a pattern of mobility following the same trend of flight take-off. Behavioral patterns of response to insecticide are, therefore, variable among strains, particularly among insecticide-resistant strains, and worth considering in resistance surveys and management programs. PMID- 19302722 TI - Trophic controls delaying foraging by termites: reasons for the ground being brown? AB - Why is the ground brown, when detritivores and decomposers have the numbers and ability to speed up the turnover of dark-coloured soil organic carbon? We consider this soil analogue to the 'green world' hypothesis measuring in the field how fast termites occupied cellulosic baits of varying quantity and quality and how predation risks by ants affect such encounters. Single baits with ants were occupied by termites later than triple baits without ants, implying that termites may spend longer searching for suitable food than feeding on it, thereby delaying decomposition rates of both chosen and neglected items. Because termites' feeding speeds up dissimilation of polymers by decomposers, such results may imply that bottom-up and top-down forces, ultimately, impair carbon processing and release from soil. We argue that the ground is brown partly because of delays imposed upon termites' use of resources by bottom-up and top down forces. PMID- 19302723 TI - Distinguishing suitable biotypes of Dactylopius tomentosus (Hemiptera: Dactylopiidae) for biological control of Cylindropuntia fulgida var. fulgida (Caryophyllales: Cactaceae) in South Africa. AB - Cylindropuntia fulgida (Engelmann) F.M. Knuth var. fulgida (Engelmann) F.M. Knuth (Cff) (Caryophyllales: Cactaceae) is native to Mexico and Arizona and was introduced into South Africa for ornamental purposes. It subsequently became highly invasive, necessitating control. The cochineal insect, Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae), was selected as a potential biological control agent based on its restricted host range among Cylindropuntia species and previous success in controlling C. imbricata (DC.) F. Knuth (Ci). Eight D. tomentosus provenances (Cholla, Cholla E, Fulgida, Mamillata, Imbricata, Tunicata U, Tunicata V and Rosea) from Cylindropuntia species in their native ranges were reared on Cff, whilst Cholla and Imbricata were also reared on Ci. Large differences were found in the development and survival of crawlers, and in the reproductive capacity of females. Three subjective categories of provenance interaction with host plants were identified based on a fitness index (FI) calculated from data relating to crawler survival, female development time and fecundity: (i) thriving (FI > or = 1) - insects had shorter developmental times, high crawler survival and highly fecund females (Cholla); (ii) surviving (FI<1 but >0) - insects had extended development times, low crawler survival and low fecundity (Imbricata, Fulgida and Mamillata); and (iii) dying (FI = 0) - insects died before or at the second instar (Rosea, Tunicata U and Tunicata V). Cholla, therefore, is highly suitable for biological control of Cff in South Africa. In addition, Cholla thrived on Cff but only survived on Ci whilst, in contrast, Imbricata thrived on Ci but only survived on Cff. This differential ability of provenances to thrive or survive on different host plants demonstrated that host adapted biotypes of D. tomentosus exist; therefore, biotypes should be taken into account when considering this species as a biological control agent of cactus weeds. PMID- 19302724 TI - Novel use of stir bar sorptive extraction (SBSE) as a tool for isolation of oviposition site attractants for gravid Culex quinquefasciatus. AB - Mosquitoes such as Culex quinquefasciatus Say (Diptera: Culicidae) are important vectors of organisms that cause disease in humans. Research into the development of effective standardized odour baits for blood-fed females (oviposition attractants), to enable entomological monitoring of vector populations, is hampered by complex protocols for extraction of physiologically active volatile chemicals from natural breeding site water samples, which have produced inconsistent results. Air entrainment and solvent extraction are technically demanding methods and are impractical for use in resource poor environments where mosquito-borne disease is most prevalent. This study reports the first use of a simple, robust extraction technique, stir bar sorptive extraction (SBSE), to extract behaviourally active small lipophilic molecules (SLMs) present in water samples collected from Cx. quinquefasciatus breeding sites in Tanzania. Extracts from a pit latrine and from a cess pool breeding site attracted more gravid Cx. quinquefasciatus in pair choice bioassays than control extracts, and coupled gas chromatography-electroantennography (GC-EAG) allowed tentative identification of 15 electrophysiologically active chemicals, including the known oviposition attractant, skatole (3-methylindole). Here, we have demonstrated, using simple pair choice bioassays in controlled laboratory conditions, that SBSE is effective for the extraction of behaviourally and electrophysiologically active semiochemicals from mosquito breeding site waters. Further research is required to confirm that SBSE is an appropriate technique for use in field surveys in the search for oviposition cues for Cx. quinquefasciatus. PMID- 19302725 TI - Genome size of Pachypsylla venusta (Hemiptera: Psyllidae) and the ploidy of its bacteriocyte, the symbiotic host cell that harbors intracellular mutualistic bacteria with the smallest cellular genome. AB - Psyllids harbor the primary symbiont, Carsonella ruddii (gamma-Proteobacteria), within the cytoplasm of specialized cells called bacteriocytes. Carsonella has the smallest known cellular genome (160 kb), lacking numerous genes that appear to be essential for bacterial life. This raises the question regarding the genetic mechanisms of the host which supports the survival of Carsonella. Our preceding analyses have indicated that some of the genes that are encoded in the psyllid genome and which are highly expressed in the bacteriocyte are of bacterial origin. This implies that psyllids acquired genes from bacteria by lateral gene transfer (LGT) and are using these genes to maintain the primary symbiont, Carsonella. To reveal the complete picture of LGT from symbiotic bacteria to the genome of psyllids, whole genome analysis of psyllids is essential. In order to assess the feasibility of whole genome analysis of the host psyllid, the genome size of the hackberry petiole gall psyllid, Pachypsylla venusta, was estimated. Feulgen image analysis densitometry and flow cytometry demonstrated that the haploid genome size of P. venusta is 0.74 pg (724 Mb), verifying the feasibility of whole genome analysis. Feulgen image analysis densitometry further revealed that bacteriocytes of P. venusta are invariably 16 ploid. This higher ploidy may be essential to facilitate the symbiotic relationship with bacteria, as it appears to be a feature common to insect bacteriocytes. These results provide a foundation for genomics-based research into host-symbiont interactions. PMID- 19302726 TI - Wing morphometry as a tool for correct identification of primary and secondary New World screwworm fly. AB - Cochliomyia hominivorax and Cochliomyia macellaria are endemic Neotropical Calliphoridae species. The former causes severe myiasis in hosts while the latter is Sarcosaprophagous, but commonly found as a second invader in wounds. Due to the morphological similarity between them and the potential losses that C. hominivorax represents for cattle breeders, the rapid and correct identification of these two species is very important. In addition to a correct identification of these species, a good knowledge of C. hominivorax biology can be helpful for designing control programs. We applied geometric morphometric methods to assess wing differences between C. hominivorax and C. macellaria and conduct a preliminary analysis of wing morphological variation in C. hominivorax populations. Canonical variate analysis, using wing shape data, correctly classified 100% of the individuals analyzed according to sex and species. This result demonstrates that wing morphometry is a simple and reliable method for identifying C. hominivorax and C. macellaria samples and can be used to monitor C. hominivorax. Both species show sexual dimorphism, but in C. hominivorax it is magnified. We suggest that this may reflect different histories of selection pressures operating on males and females. Significant differences in wing size and shape were obtained among C. hominivorax populations, with little correlation with latitude. This result suggests that wing variation is also a good morphological marker for studying population variation in C. hominivorax. PMID- 19302727 TI - Does famine have a long-term effect on cohort mortality? Evidence from the 1959 1961 great leap forward famine in China. AB - Using retrospective individual mortality records of three cohorts of newborns (1954-1958, 1959-1962 and 1963-1967) from a large national fertility survey conducted in 1988 in China, this paper examines the effect of being conceived or born during the 1959-1961 Great Leap Forward Famine on postnatal mortality. The results show strong evidence of a short-term (period) effect of the famine, caused directly by starvation or severe malnutrition during the period of the famine. After controlling for period mortality fluctuation, however, the famine born cohort does not show higher mortality than either the pre-famine or the post famine cohort. Aggregate-level cross-temporal comparisons using published cohort population counts from China's 1982 Census, 1990 Census, 1995 micro-Census, 2000 Census and 2005 micro-Census lead to the same conclusion. The relevance of these new findings for the 'fetal origins' hypothesis and the selection effect hypothesis is discussed. PMID- 19302728 TI - Male participation in pregnancy and delivery in Nigeria: a survey of antenatal attendees. AB - This was a cross-sectional study carried out on 462 pregnant women attending antenatal care in Ibadan, Nigeria. The study's aims were to assess the level of participation of Nigerian men in pregnancy and birth, the attitude of the women and likely targets for improved care delivery. Three hundred and forty-nine women (75.5%) were aware that husbands could participate in childbirth. Most women did not think it was their husbands' place to attend antenatal clinic (48.3%) or counselling sessions (56.7%). Nearly all husbands (97.4%) encouraged their wives to attend antenatal clinic - paying antenatal service bills (96.5%), paying for transport to the clinic (94.6%) and reminding them of their clinic visits (83.3%). Three hundred and thirty-five husbands (72.5%) accompanied their wives to the hospital for their last delivery, while 63.9% were present at last delivery. More-educated women were less likely to be accompanied to the antenatal clinic, while more-educated men were likely to accompany their wives. Yoruba husbands were less likely to accompany their wives, but Yoruba wives with non Yoruba husbands were 12 times more likely to be accompanied. Women in the rural centre were less likely to receive help with household chores from their husbands during pregnancy, while educated women were more likely to benefit from this. Monogamous unions and increasing level of husbands' education were associated with spousal presence at delivery. It appears that male participation is satisfactory in some aspects, but increased attendance at antenatal services and delivery would be desirable. PMID- 19302729 TI - IQ and the values of nations. AB - The origin of values and preferences is an unresolved theoretical question in behavioural and social sciences. The Savanna-IQ Interaction Hypothesis, derived from the Savanna Principle and a theory of the evolution of general intelligence, suggests that more intelligent individuals may be more likely to acquire and espouse evolutionarily novel values and preferences (such as liberalism and atheism and, for men, sexual exclusivity) than less intelligent individuals, but that general intelligence may have no effect on the acquisition and espousal of evolutionarily familiar values. Macro-level analyses show that nations with higher average intelligence are more liberal (have greater highest marginal individual tax rate and, as a result, lower income inequality), less religious (a smaller proportion of the population believes in God or considers themselves religious) and more monogamous. The average intelligence of a population appears to be the strongest predictor of its level of liberalism, atheism and monogamy. PMID- 19302730 TI - Overcoming biological barriers to in vivo efficacy of antisense oligonucleotides. AB - Antisense oligonucleotides as a therapeutic platform have been slow to progress since the approval of the first antisense drug in 1998. Recently, there have been several examples of convincing antisense interventions in animal models and promising clinical trial data. This review considers the factors determining the success of antisense oligonucleotides as therapeutic agents. In order to produce target knockdown after systemic delivery, antisense oligonucleotides must avoid nuclease degradation, reticuloendothelial-system uptake and rapid renal excretion, and extravasate to the target cell type outside the vasculature. They then must enter the target cell, and escape the endosome-lysosome pathway so as to be free to interact with the target mRNA. We consider the significance of these limiting factors based on the literature and our own experience using systemic administration of antisense oligonucleotides. PMID- 19302731 TI - Imagining intergroup contact reduces implicit prejudice. AB - Recent research has demonstrated that imagining intergroup contact can be sufficient to reduce explicit prejudice directed towards out-groups. In this research, we examined the impact of contact-related mental imagery on implicit prejudice as measured by the implicit association test. We found that, relative to a control condition, young participants who imagined talking to an elderly stranger subsequently showed more positive implicit attitudes towards elderly people in general. In a second study, we demonstrated that, relative to a control condition, non-Muslim participants who imagined talking to a Muslim stranger subsequently showed more positive implicit attitudes towards Muslims in general. We discuss the implications of these findings for furthering the application of indirect contact strategies aimed at improving intergroup relations. PMID- 19302732 TI - Effect of manipulated prestige-car ownership on both sex attractiveness ratings. AB - Previous studies have shown that male attractiveness can be enhanced by manipulation of status through, for example, the medium of costume. The present study experimentally manipulated status by seating the same target model (male and female matched for attractiveness) expressing identical facial expressions and posture in either a 'high status' (Silver Bentley Continental GT) or a 'neutral status' (Red Ford Fiesta ST) motor-car. A between-subjects design was used whereby the above photographic images were presented to male and female participants for attractiveness rating. Results showed that the male target model was rated as significantly more attractive on a rating scale of 1-10 when presented to female participants in the high compared to the neutral status context. Males were not influenced by status manipulation, as there was no significant difference between attractiveness ratings for the female seated in the high compared to the neutral condition. It would appear that despite a noticeable increase in female ownership of prestige/luxury cars over recent years males, unlike females remain oblivious to such cues in matters pertaining to opposite-sex attraction. These findings support the results of previous status enhancement of attractiveness studies especially those espousing sex differences in mate preferences are due to sex-specific adaptations. PMID- 19302733 TI - Interpersonal complementarity in responses to auditory hallucinations in psychosis. AB - OBJECTIVES: Models of interpersonal relating have proposed that people are drawn to respond to others according to a principle of complementarity, whereby perceived hostility elicits reciprocal hostility and perceived dominance elicits submission. This study examined interpersonal appraisals and responses to auditory hallucinations, to determine whether this principle predicts how patients respond to hallucinated voices. DESIGN AND METHODS: Thirty-five participants with schizophrenia or schizoaffective disorder rated their internal 'relationship' with their auditory verbal hallucinations using the Structural Analysis of Social Behaviour, in addition to completing measures of voice-related distress and depression. RESULTS: Participants were able to ascribe almost the full range of interpersonal relating behaviours to their voices reliably, with some notable exceptions for voices' 'responses' to being directly addressed. Appraisals of voice hostility very strongly predicted reciprocal hostile responses, but perceptions of voice control were only weakly associated with submission to voices. Perceived voice hostility was also associated with voice related distress and perceived voice control was associated with depression. CONCLUSIONS: Individual differences in responses to auditory hallucinations appear to reflect normal interpersonal responses to appraisals of voice hostility and affiliation. However, voice hearers do not readily submit to voice control and other factors may be of importance in determining this response. PMID- 19302734 TI - Elaborating the cry of pain model of suicidality: Testing a psychological model in a sample of first-time and repeat self-harm patients. AB - OBJECTIVES: Few studies have specifically tested the Cry of Pain model (CoP model; Williams, 2001). This model conceptualizes suicidal behaviour as a behavioural response to a stressful situation which has three components: defeat, no escape potential, and no rescue. In addition, the model specifies a mediating role for entrapment on the defeat-suicidal ideation relationship, and a moderating role for rescue factors on the entrapment-suicidal ideation relationship. This is the first study to investigate the utility of this psychological model in a sample of first-time and repeat self-harm (SH) patients. METHOD: One hundred and thirteen patients who had been admitted to hospital following an episode of SH (36 first-time, 67 repeat) and 37 hospital controls completed measures of defeat, entrapment/escape potential, rescue (social support and positive future thinking), as well as depression, anxiety, and suicidal ideation. RESULTS: Analyses highlighted differences between the three participant groups on all of the CoP variables. Hierarchical regression analysis confirmed that total entrapment and internal entrapment mediated the relationship between defeat and suicidal ideation, whilst impaired ability to think positively about the future (but not social support) moderated the relationship between total and internal entrapment and suicidal ideation. CONCLUSIONS: The findings provide further empirical support for the CoP Model. The findings are discussed in relation to theory and practice and we recommend that the findings are replicated within a prospective design. PMID- 19302735 TI - New strategy for diagnosis and treatment of gynecological cancer. PMID- 19302737 TI - Predictors and clinical significance of the positive cone margin in cervical intraepithelial neoplasia III patients. AB - BACKGROUND: Conization is being widely accepted for diagnosis and treatment of cervical intraepithelial neoplasia (CIN). There is controversy as to which factors are most predictive of a positive cone margin and the clinical significance of it. We conducted this study to identify the predictive factors and to evaluate the clinical significance of a positive cone margin in CIN III patients. METHODS: A retrospective review was conducted of 207 patients who had undergone conization due to CIN III from January 2003 to December 2005 at Peking Union Medical College Hospital. Of these, 67 had a subsequent hysterectomy. Univariate and multivariate analysis were utilized to define the predictive factors for a positive cone margin, and to compare the pathologic results of conization with subsequent hysterectomy. RESULTS: One hundred and fifty-one (72.9%) were margin free of CIN I or worse, 37 (17.9%) had CIN lesions close to the margin and 19 (9.2%) had margin involvement. A total of 56 cases (27.1%) had positive cone margins (defined as the presence of CIN at or close to the edge of a cone specimen). Univariate analysis showed that the parity, cytological grade, multi-quadrants of CIN III by punch biopsy, gland involvement, as well as the depth of conization were significant factors correlated with a positive cone margin (P < 0.05). However the age, gravidity, grade of dysplasia in punch biopsy, as well as the cone methods were not significantly correlated (P > 0.05). Multivariate analysis revealed that the cytological grade (OR = 1.92), depth of conization (OR = 2.03), parity (OR = 3.02) and multi-quadrants of CIN III (OR = 4.60) were significant predictors with increased risk for positive margin. The frequency of residual CIN I or worse in hysterectomy specimens was found to be 55.6% (20/36) in patients who were margin free, 71.4% (15/21) in patients with CIN occurring close to margin, and 80.0% (8/10) in patients with margin involvement. The frequency of residual CIN III or worse was found to be 13.9% (5/36), 23.8% (5/21) and 50.0% (5/10) respectively in different groups. CONCLUSIONS: Cytological grade, depth of conization, parity and multi-quadrants of CIN III in punch biopsy were significant factors with increased risk in predicting a positive cone margin. Margin status of conization did not mean the presence or absence of CIN, but rather the varied frequency of residual CIN in specimens of subsequent hysterectomy. In view of this fact, it is suggested that the margin status of conization be a valuable surrogate marker for clinical management of CIN III. PMID- 19302738 TI - Effect of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry on identifing biomarkers of endometriosis. AB - BACKGROUND: Endometriosis is a common gynecological disease. This study aimed to screen proteins that were expressed differently in patients with endometriosis versus normal controls using proteomic techniques, surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). METHODS: Protein chip SELDI-TOF-MS combines the advantages of microarray and mass spectrometry, and can screen latent markers in sera of patients with endometriosis. Serum samples from patients and normal volunteers were analyzed by SELDI-TOF-MS. RESULTS: After comparing the serum protein spectra of 36 patients with 24 normal controls, 24 differently expressed potential biomarkers (P < 0.01) were identified. Using Biomarker Pattern software, we established a tree model of the 60 serum protein spectra. When using the three biomarkers to classify the samples, the sensitivity for diagnosing endometriosis was 91.7%, specificity was 95.8%, and coincidence rate was 93.3%. Then we used serum samples from 12 patients and 8 normal controls to validate the tree model and report the sensitivity for diagnosing endometriosis was 91.7%, specificity was 75%, and coincidence rate was 85%. CONCLUSIONS: SELDI-TOF-MS may be a useful tool in high risk population screening for endometriosis. The identification and application of the biomarkers need to further study. PMID- 19302739 TI - Clinical assessment for three routes of hysterectomy. AB - BACKGROUND: Hysterectomy is a very common surgery in gynecology. Ideal surgery for hysterectomy is microinvasive with few complications. There are three major routes of hysterectomy that are currently used. The aim of this study was to identify the differences of peri-operative outcome among the patients who underwent the three different approaches. METHODS: One hundred and one women undergoing hysterectomy for myoma had the procedure performed by laparoscopic assisted vaginal hysterectomy (LAVH), total vaginal hysterectomy (TVH) or total abdominal hysterectomy (TAH) in a randomized study. We compared the course of peri-operative and post-operative outcome for the three different approaches. RESULTS: were evaluated by linear regression analysis, Fisher's exact test and Student's t test for independent samples. RESULTS: The operation time among the three procedures was not significantly different (P > 0.05). The amount of blood loss in the TVH group was less than in the LAVH and TAH groups (P < 0.05). The pain score 3 hours after operation in the LAVH group was significantly lower than in the TAH and TVH groups (P < 0.001). The pain scores in the LAVH and TVH groups were lower than in the TAH group at 24 and 48 hours after operation (P < 0.01). The women who underwent LAVH and TVH had a shorter hospitalization stay (P < 0.001). The highest body temperature after operation in the TAH group was higher than that in LAVH and TVH groups (P < 0.001). CONCLUSIONS: LAVH and TVH are better procedures for women requiring hysterectomy. The peri-operative and post operative courses of TVH are better than LAVH, excluding the pain score 3 hours after operation. Vaginal hysterectomy is the most cost-effective approach but the final choice for the route of hysterectomy can depend on many factors such as gynecological disease, patients' health status and experiences of the gynecologist. PMID- 19302740 TI - Effects of estrogen and tibolone on bladder histology and estrogen receptors in rats. AB - BACKGROUND: Estrogen deficiency causes atrophic changes within the urogenital tract, and is associated with urinary symptoms. The purpose of this study was to investigate the effects of estrogen and tibolone on bladder histology, and the changes of estrogen receptor alpha and beta (ERalpha and beta) protein expression in the detrusor muscle. METHODS: Forty female rats were separated into four groups of ten each. They received a sham operation (Sham), ovariectomy (Ovx), ovariectomy plus estrogen replacement (Ovx + E), or ovariectomy plus tibolone treatment (Ovx + T). After 12 weeks each rat was anesthetized and the bladders were removed. The bladders' ultra structure, collagen fiber (CF) to smooth muscle (SM) ratio and ER subtypes were studied. Statistical analyses were performed using the one-way analysis of variance test. RESULTS: Ovx resulted in significant degeneration in bladder ultra structure; however, estrogen and tibolone reversed those changes. Ovx increased the CF/SM ratio, estrogen and tibolone resulted in an increase. Two estrogen receptors (ERs) were expressed in the bladder detrusor, with ERbeta the main subtype. Ovx resulted in up-regulation of ERalpha and down regulation of ERbeta. With estrogen and tibolone treatment, ERbeta showed a significant increase but ERalpha showed no significant difference compared with Ovx. CONCLUSIONS: Estrogen deficiency deteriorates bladder ultra structure and histology. Supplementary estrogen can improve bladder function which may be due to inhibition of collagen hyperplasia and increased SM density. ERbeta has an important role in mediating estrogen function in the bladder. Tibolone has a mild estrogenic action and has an effect on bladder function and structure to some degree. PMID- 19302741 TI - Influential and prognostic factors of small for gestational age infants. AB - BACKGROUND: Small for gestational age (SGA) infants are associated with a high rate of oligohydramnios, stillbirth and cesarean delivery. Among SGA patients there is a higher risk of neonatal complications, such as polycythemia, hyperbilirubinemia, and hypothermia. Additionally, the SGA infant is prone to suffer from major neurologic sequelae, as well as cardiovascular system disease, in later life. Proper monitoring and therapy during pregnancy are, therefore, of utmost importance. The present study aimed to investigate the influential and prognostic factors of SGA infants. METHODS: From January 2001 to June 2007, a total of 55 SGA neonatal infants were included in a study group. All were born at Peking Union Medical College Hospital, with regular formal antenatal examinations. In addition, a total of 122 cases of appropriate for gestational age (AGA) infants were born at the same time and were registered into a control group. All cases were singleton pregnancies with detailed information of the maternal age, gravidity, parity, maternal height and weight, complications, uterine height and abdominal circumference, results from transabdominal ultrasonography between 32 - 38 gestational weeks, pregnancy duration, delivery manner, placenta, umbilical cord, and neonatal complications. RESULTS: Significant differences were observed in placenta weight and neonatal malformations between the study and control groups. Multivariate analysis revealed increased parity, maternal hyperthyroidism and hyperthyroidism history as risk factors. Fetal abdominal circumferences less than 30 and 32 cm at 32 - 38 gestational weeks respectively, as determined by ultrasonography, resulted in a Youden index of 0.62. CONCLUSIONS: SGA infants were associated with a greater risk of smaller placentas and infant malformations. Increased parity, maternal hyperthyroidism, and a hyperthyroid history were risk factors for SGA infants. Fetal abdominal circumference less than 30 cm at 32 gestational weeks and less than 32 cm at 38 weeks, as determined by ultrasonography, was considered an effective index for SGA. PMID- 19302742 TI - Relationship between expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4(+) T cells and spontaneous abortion in mice. AB - BACKGROUND: Previous studies have shown that local immune cells in the feto maternal interface are recruited from peripheral blood, and that chemokines and their receptors play an initial and key role in this recruitment process. In this study, we aimed to determine whether spontaneous abortion is associated with the expression of chemokine receptors CCR3, CCR5, and CXCR3 on CD4(+) T cells. METHODS: Peripheral blood, spleen, and thymus were collected from the spontaneous abortion mouse model CBA/JxDBA/2 (SA group, n = 14), the normal pregnant mouse model CBA/JxBALB/c (NP group, n = 13), and normal non-pregnant CBA/J mice (NNP group, n = 11). The number of chemokine receptors CCR3, CCR5, and CXCR3 expressed on CD4(+) T cells was measured by double-label flow cytometry (FCM) method. RESULTS: In peripheral blood, the SA group had significantly lower CCR3 expression (P < 0.01) and higher CCR5 and CXCR3 expression (P < 0.01) on CD4(+) T cells than did the NP group. But comparing these chemokines between the SA and NNP groups, there was no significant difference (P > 0.05). In spleen, the SA group expressed significantly lower CCR3 expression (P < 0.01) and higher CCR5 and CXCR3 expression (P < 0.05) on CD4(+) T cells than did the NP group. When compared with the NNP group, the SA group had significantly higher CCR3 expression (P < 0.01), but was not statistically different with regards to the other two chemokines (P > 0.05). In thymus, the SA group had significantly lower CCR3 expression (P < 0.05) and higher CXCR3 expression (P < 0.05) on CD4(+) T cells than the NP group, with no significant difference in CCR5 expression (P > 0.05). Compared with the NNP group, the SA group had higher CCR3 expression (P < 0.01), but there was no statistical difference in CXCR3 and CCR5 expression (P > 0.05) between the two groups. CONCLUSION: The abnormal expression of CCR3, CCR5 and CXCR3 on CD4(+) T cells may play an important role in the pathogenesis of spontaneous abortion. PMID- 19302743 TI - Effect of coronary bifurcation angle on clinical outcomes in Chinese patients treated with crush stenting: a subgroup analysis from DKCRUSH-1 bifurcation study. AB - BACKGROUND: Bifurcation angles may have an impact on the clinical outcomes of crush stenting. We sought to compare high (> or = 60 degrees ) with low (< 60 degrees ) bifurcation angle in patients who underwent either classical or double kissing (DK) crush stenting for bifurcation lesions from the DKCRUSH-1 data base. METHODS: There were 212 patients with 220 lesions, some with low-angle (n = 138) and some with high-angle (n = 74). Angiography was indexed at 8-month after procedure. Primary endpoint was the occurrence of major adverse cardiac events (MACEs), defined as cardiac death, myocardial infarction and target lesion revascularization (TLR). Secondary endpoint included late lumen loss, the rate of restenosis, and final kissing balloon inflation (FKBI). RESULTS: At 8 months, clinical follow-up was 100%; angiographic follow-up was 75% in the low-angle group and 83.3% in the high-angle group. There were no significant differences in the FKBI between the high-angle group (91.43%) and the low-angle group (82.39%). In the high angle group, there was a significant difference in contrast volume used (P = 0.005) but no significant difference in acute gain, minimum lumen diameter (MLD), late loss and diameter stenosis in the pre-bifurcation segment, post-bifurcation segment or side branch. When lesions were assigned into with-(n = 133) and without-FKBI (n = 42), significant side-branch late loss was seen in the group without-FKBI ((0.65 +/- 0.49) mm vs (0.47 +/- 0.62) mm, P = 0.02), with a resultant greater restenosis rate (37.68% vs 18.32%, P = 0.001). No difference was detected in the MACE free survival rate between the high and low angle groups (82.39% vs 82.36%, P = 0.84). The rate of stent thrombosis tended to be higher in the lower-angle group although there was no significant difference (P = 0.38). The TLR free survival rate was 87.2% in the with-FKBI group vs 73.5% in the without-FKBI group (P = 0.001). Cox regression analysis showed that the independent predictors for target vessel revascularization were the side branch stent MLD post stenting (hazard ratios (HR) 1.028, 95% CI 2.357 - 16.233, P = 0.002), lack of FKBI (HR 4.910, 95% CI 4.706 - 8.459, P = 0.001) and unsatisfactory kissing (HR 3.120, 95% CI 2.975 - 5.431, P = 0.001). CONCLUSIONS: Bifurcation angles do not influence the clinical outcome of crush stenting. Successful final kissing balloon inflation, regardless of bifurcation angles, can predict TLR. PMID- 19302745 TI - Mutation of plakophilin-2 gene in arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is one of the leading causes of sudden cardiac death. Recent studies have shown that ARVC, which is an inheritable genetic change, results from mutations in genes encoding desmosomal proteins. Plakophilin-2 is an important component of the desmosome. Because the full range of genetic variations related to ARVC is unknown and no related studies of the Chinese population have been reported, we aimed to investigate the genetic variation of plakophilin-2 in ARVC patients from the Southern Region of China. METHODS: Genomic DNA was isolated from peripheral blood samples of all 34 ARVC patients, who were screened through a clinical evaluation. They were used to detect variations in the sequences of the plakophilin-2 genes by polymerase chain reaction amplification in combination with direct sequencing. RESULTS: In exon-1 of the plakophilin-2 gene, a deletion mutation (c.145_148 del GACA) was found in one family pedigree. The mutation was also found in exon-2, 4, and 11 of the plakophilin-2 gene. The QT interval dispersion of the ECG was considerably longer in the mutation group than in the non-mutation group of ARVC patients, and this result was statistically significant (P < 0.05). CONCLUSION: We discovered a plakophilin-2 mutation that prolongs the QT interval dispersion in the southern Chinese ARVC population. PMID- 19302746 TI - Clinical assessment of awake endotracheal intubation using the lightwand technique alone in patients with difficult airways. AB - BACKGROUND: There is few study to determine whether the use of the lightwand technique alone could achieve effective, safe and successful awake endotracheal intubation (ETI), therefore we designed a prospective clinical study to systematically evaluate the feasibility, safety and efficacy of awake ETI using the lightwand alone in patients with difficult airways. METHODS: Seventy adult patients with difficult airways were enrolled in this study. After the desired sedation with fentanyl and midazolam, airway topical anesthesia was performed with 9 ml of 2% lidocaine, which were in order sprayed in three aliquots at 5 minutes intervals into the supraglottic (two doses) and laryngotracheal areas (one dose) using a combined unit of the lightwand and MADgic atomizer. After airway topical anesthesia, awake ETI was performed using a Lightwand. Subjective assessments by patients and operators using the visual analogue scores (VAS), and objective assessments by an independent investigator using patients' tolerance and reaction scores, coughing severity, intubating conditions and cardiovascular variables were taken as the observed parameters. RESULTS: Of 210 airway sprays, 197 (93.8%) were successfully completed on the first attempt. The total time for airway spray was (14.6 +/- 1.5) minutes. During airway topical anesthesia, the average patients' tolerance scores were 1.7 - 2.3. After airway topical anesthesia, the mean VAS for discomfort levels that the patients reported was 6.5. Also airway topical anesthesia procedure was rated as acceptable and no discomfort by 94.3% of patients. The lightwand-guided awake ETI was successfully completed on first attempt within 29 seconds in all patients. During awake ETI, patients' reaction and coughing scores were 1.9 and 1.6, respectively. All patients exhibited excellent or acceptable intubating conditions. Cardiovascular monitoring revealed that changes of systolic blood pressure and heart rate at each stage of airway manipulations were less than 20% of baseline values. The postoperative follow-up showed that 95.7% of patients had no recall or slight memories of all airway instrumentation. The incidence of postoperative mild airway complications was 38.6%. CONCLUSION: Alone use of the lightwand technique can achieve effective, safe and successful awake ETI in patients with difficult airways. PMID- 19302747 TI - GATA4 and NKX2.5 gene analysis in Chinese Uygur patients with congenital heart disease. AB - BACKGROUND: Congenital heart disease (CHD) is the most common developmental anomaly in newborns. The germline mutations in GATA4 and NKX2.5 genes have been identified as responsible for CHD. The frequency of GATA4 and NKX2.5 mutations in Chinese Uygur patients with CHD and the correlation between their genotype and CHD phenotype are unknown. METHODS: We examined the coding region of GATA4 and NKX2.5 genes in 62 Chinese Uygur patients with CHD and 117 Chinese Uygur individuals as the controls by denaturing high performance liquid chromatography (DHPLC) and sequencing. RESULTS: Two heterozygous missense mutations of c.1220C > A and c.1273G > A in GATA4 gene, which cause the amino acid residue changes of P407Q and D425N in GATA4, were found in a patient with tetralogy of Fallot and a patient with ventricular septal defect, respectively. The two patients did not have atrioventricular conduct defects or non-cardiac abnormalities. The two mutations are expected to affect the protein function. There were no reported NKX2.5 mutations in the patients. CONCLUSION: Our results provided the primary data on CHD phenotype associated with GATA4 mutation in the Chinese Uygur population. PMID- 19302748 TI - Impact of human leukocyte antigen matching and recipients' panel reactive antibodies on two-year outcome in presensitized renal allograft recipients. AB - BACKGROUND: Renal transplantation in sensitized candidates remains a highly significant challenge worldwide. The production of panel reactive antibody (PRA) against human leukocyte antigen (HLA) is a major risk factor in presensitized recipients. The aim of this study was to evaluate the impact of HLA matching and recipients' PRA on two-year outcome in presensitized renal allograft recipients. METHODS: We determined the percentage of panel reactivity and specificity of anti HLA immunoglobulin (Ig) G antibodies in 73 presensitized renal allograft recipients compared with 81 unsensitized recipients (control group). HLA genotyping of both recipients and corresponding donors was performed by PCR with sequence-specific primers (PCR-SSP). We analyzed the factors influencing the early graft outcome (two-year rejection rates and survival rates of the grafts), including HLA mismatching, class and degree of panel reactivity, and target antigen of donors. RESULTS: Presensitized recipients had a worse two-year outcome than unsensitized recipients (P = 0.019 for rejection rate, P = 0.01 for survival rate). The difference in number of HLA-mismatched alleles with either 6-antigen matching (Ag M) standard or amino acid residue matching (Res M) standard was not significant between the rejection and non-rejection groups of presensitized recipients or between the graft survival group and graft loss group. Compared with the control group, recipients with both PRA-I and PRA-II antibodies had a significantly worse two-year outcome (P = 0.001 for rejection rate, P = 0.002 for survival rate). The two-year outcomes of the peak PRA >/= 50% group and its subgroup, at-transplant PRA > or = 50% group, were significantly worse compared with the control group (P = 0.025 and P = 0.001 for rejection rate, P = 0.043 and P = 0.024 for survival rate). The rejection rates of the at-transplant target antigen positive group and its subgroup, HLA-I target antigen positive group, were significantly higher than the control group (P = 0.001 and P = 0.001), target antigen negative group (P = 0.003 and P = 0.001), and peak target antigen positive with negative at-transplant target antigen group (P = 0.024 and P = 0.002). Two-year graft survival rates of the target antigen positive group and HLA-I target antigen positive group were significantly lower than the control group (P = 0.012 and P = 0.001). The two-year outcome of target antigen unknown group was similar to that of the target antigen positive group. Presensitized recipients with pre-transplant plasmapheresis or immunoadsorption (PRA prepared group) had a better but non-significant two-year outcome than the control group. However, the PRA unprepared presensitized recipients were different to the control group (P = 0.004 for rejection rate and P = 0.005 for survival rate). Hyperacute rejection (HR) occurred in three recipients with positive HLA-I target antigen and without mismatch according to Res M and in one case with positive PRA II (for an unknown target antigen). No HR occurred in eight cases with positive HLA-II target antigens. CONCLUSIONS: Pre-transplant PRA preparations might improve the access of presensitized patients to renal donors. Avoiding antigen positive donors remains a fundamental measure in preventing HR and early rejections. PMID- 19302749 TI - Continuous epidural block of the cervical vertebrae for cervicogenic headache. AB - BACKGROUND: Cervicogenic headache (CEH) is caused by a structural abnormality in the cervical spine. Available treatments for CEH include medical therapy, local botulinum toxin injection, cervical epidural corticosteroid injection, and surgery. The objective of this study was to investigate the safety and efficacy of a continuous epidural block of the cervical vertebra. METHODS: Medical records were retrospectively analyzed for 37 patients diagnosed with CEH treated by a continuous epidural block of the cervical vertebra with lidocaine, dexamethasone, and saline (5 ml/min) for 3 - 4 weeks and triamcinolone acetonide 5 mg once weekly for 3 - 4 weeks. Pain was measured via the visual analogue scale (VAS) in combination with quality of life assessment. Outcome measures were patient reported days with mild or moderate pain, occurrence of severe pain, and the daily oral dosages of non-steroidal anti-inflammatory drug use (NSAID). RESULTS: In the 3 months immediately preceding placement of the epidural catheter, the mean number of days with mild or moderate pain was 22.0 +/- 4.3. The mean occurrence of severe pain was (3.20 +/- 0.75) times and the mean oral dosage of NSAID was (1267 +/- 325) mg. During the first 6 months after epidural administration of lidocaine and corticosteroids, the mean number of days with mild or moderate pain, the mean occurrence of severe pain, and the mean daily oral dosages of NSAIDs were significantly decreased compared to 3-month period immediately preceding treatment (P < 0.01). By 12 months post-treatment, no significant difference in these three outcome measures was noted. CONCLUSIONS: Continuous epidural block of the cervical vertebra for patients with CEH is effective for at least six months. Further research is needed to elucidate mechanisms of action and to prolong this effect. PMID- 19302750 TI - Mitochondrial DNA 4977-bp deletion correlated with reactive oxygen species production and manganese superoxidedismutase expression in gastric tumor cells. AB - BACKGROUND: Mitochondrial DNA 4977-bp deletion (DeltamtDNA(4977)) was reported in many human neoplasia. However, its biological significance remains to be evaluated and the molecular mechanism needs to be investigated. In this study, we analyzed the frequency of DeltamtDNA(4977) in gastric cancer (GC) cell lines and tissues, as well as reactive oxygen species (ROS) contents and manganese superoxide dismutase (MnSOD) expression levels in GC cell lines to explore its biological significance and molecular mechanism. METHODS: Semi-quantitative PCR and real-time PCR were used to detect the incidence of DeltamtDNA(4977) in 13 GC cell lines and 272 human gastric tissues (108 GC specimens and the respective adjacent normal tissues, and 56 normal gastric mucosa from non-cancer patients). We further identified intracellular ROS production by flow cytometry and MnSOD expression by semi-quantitative reverse transcription-PCR (RT-PCR) and Western blotting. Statistical analyses were carried out using the Logistic regression analysis and Kaplan-Meier method. RESULTS: Based on our earlier study, we optimized the PCR amplification condition by reducing the cycle number. In this study, we systematically documented the high incidence of DeltamtDNA(4977) in GC cell lines (10/13, 76.9%), GC tissues (86/108, 79.6%), matched normal tissues (73/108, 67.6%), and normal gastric mucosa of non-cancer patients (29/56, 51.8%). A significantly higher incidence of mutated DeltamtDNA(4977) was observed in GC tissues with respect to the adjacent normal tissues (79.6% vs 67.6%, P = 0.045), and they were both higher than that in normal controls (P < 0.05). Most importantly, we linked the DeltamtDNA(4977) mutations with the expression level of MnSOD and ROS contents. The cell lines containing lower expression level of MnSOD was found to have generally higher frequent DeltamtDNA(4977) and more ROS. CONCLUSION: The decreased anti-oxidative ability, which leads to increased ROS contents, is correlated with the mtDNA damage during gastric carcinogenesis. PMID- 19302751 TI - Alteration of gene expression during nasopharyngeal carcinogenesis revealed by oligonucleotide microarray after microdissection of tumor tissue and normal epithelia from nasopharynx. AB - BACKGROUND: Microarray and microdissection techniques were being used for many applications to study the carcinogenesis of some human tumors. But seldom studies had hitherto combined these two techniques to study carcinogenesis mechanism of nasopharyngeal carcinoma (NPC). To identify a set of genes involved in the carcinogenesis and development of NPC, we used the microdissected homogeneous NPC tissue cells and the pure normal epithelium pillar cells to construct the whole human genome expression profiles. METHODS: We preserved the tissue samples from nasopharynx of 18 patients (including 13 samples of NPC and 5 samples of normal or inflammatory mucous tissue samples from nasopharynx) in RNAlater Stabilization Reagent. The tissue samples were microdissected to harvest the homogeneous tissue cells, then total RNA was isolated from them. The sufficient antisense RNA (aRNA) was amplified from these total RNA. HG-U133.Plus.2.0 GeneChip was used to construct the human whole genome expression profiling of each sample. Differential patterns of expression of genes correlated with the carcinogenesis, classification and progression of NPC were identified with comparing the expression profiling data respectively in leave one out cross-validation analysis. Correlation between aRNA expression measured by the microarrays and semi-quantitative reverse transcription polymerase chain reaction (sqRT-PCR) were also ascertained, and found that hybridization results were validated in all of the 18 patients. RESULTS: Differential patterns of expression of 127 genes correlated with the carcinogenesis (A P value less than 0.001 with the 2-fold differentiated expression between case group and control group) of the NPC were filtered. The top most up-regulated and down-regulated 8 genes by the way of permutation test were also selected and listed in the paper. Expression of genes E2F6 and TSPAN-1 was identified using aRNA by sqRT-PCR and showed that there was significant difference between the average value of case groups and that of control group respectively (t = 2.170, df = 16, P = 0.045 and t = -2.946, df = 16, P = 0.009). CONCLUSIONS: We had identified some genes which could be the molecular marker during the carcinogenesis and the development of the NPC. The genes which selected from the different subgroups seemed to be implicated for the diagnosis,classification, and progression of NPC, and provided important insights into their underlying biology. PMID- 19302752 TI - Passive cigarette smoking induces inflammatory injury in human arterial walls. AB - BACKGROUND: Epidemiological studies have shown that both active and passive cigarette smoking increase the risk of atherosclerosis. But very little is known about the biological processes induced by passive cigarette smoking that contribute to atherosclerosis. We observe the expression of a few of biological and inflammatory markers in human arterial walls in vitro which were treated with the second-hand smoke solution (sidestream whole, SSW), and discuss the possible mechanism of inflammatory injury induced by second-hand smoke. METHODS: The biological markers (platelet endothelial cell adhesion molecule-1, PECAM-1; alpha smooth muscle actin, alpha-SMA; collagen IV, Col IV) and inflammatory markers (vascular cell adhesion molecule-1, VCAM-1; monocyte chemoattractant protein-1, MCP-1; interleukin-8, IL-8) of human aortal wall were tested by immunofluorescence staining. The levels of MCP-1 and IL-8 mRNA expression were detected by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: No distinct difference was observed between SSW and the control group on the expression of biological markers as assessed by the light microscope. But the inflammatory markers VCAM-1, MCP-1 and IL-8 on the subendothelial layer and smooth muscle cell layers, which are near the endothelium of arterial wall, were strongly stained in the SSW group compared with the control group. Their fluorescence intensities in the 1:40 SSW group (VCAM-1: 0.35 +/- 0.04, MCP-1: 0.34 +/- 0.05, IL-8: 0.37 +/- 0.05) and the 1:20 SSW group (VCAM-1: 0.40 +/- 0.04, MCP-1: 0.52 +/- 0.09, IL-8: 0.51 +/- 0.07) were significantly stronger than the control group (VCAM-1: 0.12 +/- 0.04, MCP-1: 0.06 +/- 0.02, IL-8: 0.24 +/- 0.03) by semi-quantitative analysis of immunofluorescence (P < 0.001 vs control). MCP-1 mRNA expression in the 1:40 SSW (0.15 +/- 0.04) and the 1:20 SSW (0.19 +/- 0.06) group was significantly higher than in the control group (0.09 +/- 0.03) (P < 0.05, P < 0.01 vs control); IL-8 mRNA expression in the 1:40 SSW (0.64 +/- 0.12) and 1:20 SSW (0.72 +/- 0.13) groups was also significantly higher than that in the control group (0.49 +/- 0.13) (P < 0.05, P < 0.01 vs control) by RT-PCR. CONCLUSIONS: It is implied that a second-hand smoke solution induces the inflammatory reaction of the arterial wall by release of inflammatory factors even though there is no distinct structural change on the arterial walls under light microscope, indicating that passive cigarette smoking is related to inflammatory injury in human arterial wall and could be closely related to the early inflammatory stage of atherosclerosis. PMID- 19302753 TI - Severity of sepsis is correlated with the elevation of serum high-mobility group box 1 in rats. AB - BACKGROUND: Sepsis is a leading cause of death in the intensive care units. The late inflammatory cytokine, high-mobility group box 1 (HMGB1), plays a critical role in sepsis. In the present study, we investigated the association between the serum HMGB1 levels and the severity of organ injury in the lipopolysaccharide induced sepsis in rats. METHODS: To produce an animal model of sepsis with different degree of organ injury, animals were treated with three different doses of lipopolysaccharide (4, 8 and 16 mg/kg), and the animals in control group were treated with the same volume of the vehicle (saline). The levels of serum HMGB1 were measured at 0, 2, 4, 8, 16, 24, 32 and 48 hours after lipopolysaccharide (LPS) or vehicle injection, meanwhile the biochemical and histopathological indicators for the severity of organ injury were assessed. RESULTS: The level of HMGB1 had a positive, high correlation with the abnormal changes of serum cardiac troponin I, alanine aminotransferase, aspartate aminotransferase, creatinine and blood urea nitrogen, as well as the pathologic scores of heart, lung, liver and kidney. CONCLUSIONS: The level of serum HMGB1 is highly correlated with the severity of sepsis in rats, suggesting that HMGB1 could serve as a valuable adjunct in the diagnosis and management of sepsis. PMID- 19302754 TI - Effects of ketamine and midazolam on morphology of dendritic spines in hippocampal CA1 region of neonatal mice. AB - BACKGROUND: It is a common phenomenon that children experience multiple general anesthesias in clinical practice, which raises the question whether repeated exposure to general anesthetics would interfere with the development of the central nervous system of children. The present study was designed to evaluate the effects of repeated treatment with ketamine or midazolam on postnatal dendrite development by examining the morphology of the dendritic spines of the pyramidal neurons in the hippocampal CA1 region in mice. METHODS: The transgenic green fluorescent protein-M line (GFP-M) mice were used in this study. Ketamine (100 mg/kg), midazolam (50 mg/kg) or saline (10 ml/kg) was administered intraperitoneally once a day on consecutive days from postnatal day 8 (P8) to postnatal day 12 (P12). At postnatal day 13 (P13) and postnatal day 30 (P30), the density and length of the apical dendritic spines of the pyramidal neurons in the hippocampal CA1 region were examined under a confocal microscope. RESULTS: At P13, for both the ketamine group and the midazolam group, the dendritic spines were found with a comparatively lower density and longer average length than in the control group. At P30, no significant difference in the density or average length of dendritic spines was found between the anesthetic group and control group. CONCLUSIONS: This study indicated that repeated exposure to ketamine or midazolam in neonatal mice impaired dendritic spine maturation immediately afterwards, but this influence seemed to disappear during further postnatal development. PMID- 19302755 TI - Effect of a conserved peptide derived from Kunitz domain of hepatitis B virus x protein on the cell cycle and apoptosis of HepG2 cells via the proteasome pathway. AB - BACKGROUND: Hepatitis B virus (HBV) x protein (HBx) in HepG2 cells causes a moderate decrease in proteolysis activity of the proteasome. A highly conserved Kunitz-type serine protease inhibitor domain within 154 amino acid residues of HBx has been identified. In this study, a peptide chain derived from the Kunitz domain (PKD) was used to study its effect on the cell cycle and apoptosis of HepG2 cells, and investigated the function of PKD on the activities of proteasomes and AAA-ATPase p97, which involves in the ubiquitin-proteasome protein degradation pathway. METHODS: The PKD peptide (Phe-Val-Leu-Gly-Gly-Cys Arg-His-Lys) was chemically synthesized. MTT assays were used to determine the effects of PKD on HepG2 cell growth. Mouse anti-p97 antibody was developed for Western blotting to detect the expression of p97. ATPase activity of proteasomes was measured using a colorimetric assay. Peptidase activities of proteasomes were analyzed with various peptidase-specific fluorogenic peptide substrates. Flow cytometry was used to determinate cell cycle phase and apoptosis. RESULTS: Viability of HepG2 cells decreased in a PKD-dose-dependent manner. Cells exhibited significant cytotoxicity in the presence of 15 mmol/L of PKD. Western blotting analysis showed that expression of p97 was suppressed in HepG2 cells treated with PKD compared to untreated cells. The ATPase activity of proteasomes from immunoprecipitates of HepG2 cells pretreated with PKD was apparently decreased. Chymotryptic activity of proteasomes in HepG2 cells was significantly inhibited by 10 mmol/L PKD; tryptic activity and peptidylglutamyl peptide hydrolase activity of proteasomes were less inhibited by PKD than chymotryptic activity. The cell cycle phase of HepG2 cells treated with PKD for 36 hours was blocked largely at the G(0)-G(1) phase, while untreated control cells were mainly in S phase. PKD also significantly induced apoptosis. CONCLUSIONS: The peptide derived from Kunitz domain of HBx protein induces HepG2 cell growth arrest and apoptosis, which may result from down-regulation of p97 expression, and decrease of both the ATPase and chymotryptic activities of proteasomes. PMID- 19302756 TI - Issues on the selection of non-inferiority margin in clinical trials. AB - OBJECTIVE: The determination of non-inferiority margin is an important and confusing issue which directly influences the acceptability of a new medication. We reviewed the published literature, International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) Guidelines and Committee for Proprietary Medicinal Products (CPMP) papers on the selection of non-inferiority margin and the corresponding statistical tests in clinical trials, in order to provide practical recommendations and suggestions for establishing reference criteria for the non-inferiority margin in China. Data sources The literature on the selection of a non-inferiority margin and statistical tests was mainly extracted from relevant English articles on non inferior clinical trials published from 1990 to 2007. The starting point (1990) was chosen due to lack of such papers published prior to 1990. This literature was searched via PubMed, Medline and Chinese Knowledge Information (CNKI). ICH guidelines and CPMP papers were downloaded from their official websites. The keywords "clinical trial", "non-inferiority" and "non-inferiority margin" were used. Study selection Forty-three original articles and critical reviews, ICH E10 guideline and CPMP papers were selected. RESULTS: The non-inferiority testing with treatment difference and ratio are commonly used, where the non-inferiority margin is determined with and without historical data. Traditionally, this margin is treated as a fixed value, while developed methods take the variation into account in the determination of this margin, on which the test depends is more convincing. The mixed margin consisting of a margin based on treatment difference and a margin based on treatment ratio can exactly control the type I error at the desirable level and obtain a better power. In this review, we also provide some recommendations and suggestions for the selection of the non-inferiority margin in the western countries and China. CONCLUSIONS: Besides the magnitude and variability of the standard control and placebo effects, the determination of the margin depends on other factors, such as, the safety profiles of the two comparative therapies, method of administration and product cost. Hence the margin should be made specifically for a certain case. Reference criteria for different medications should be given to avoid the arbitrary conclusion. PMID- 19302757 TI - Construction and expression of a bicistronic vector containing human bone morphogenetic protein 2 and vascular endothelial growth factor-165 genes in vitro. PMID- 19302758 TI - Leiomyomatosis peritonealis disseminata associated with endocervicosis. PMID- 19302759 TI - High-frequency jet ventilation for right upper pulmonary lobe sleeve resection. PMID- 19302760 TI - Value of transcranial motor evoked potentials during spinal operations. PMID- 19302761 TI - Olanzapine as treatment for children and adolescents with Tourette's syndrome. PMID- 19302762 TI - Psychosis in children with velocardiofacial syndrome (22q11.2 deletion syndrome). AB - Velocardiofacial syndrome, now known as 22q11.2 deletion syndrome (22qDS), is estimated to affect more than 700 children born in the United States each year. Some clinical studies have found increased rates of schizophrenia in adults with 22qDS. However, these studies have been limited by small sample size and possible ascertainment bias. The psychiatric disorders most commonly reported in children and adolescents with 22qDS have been attention-deficit/hyperactivity disorder, oppositional defiant disorder, anxiety disorders, and major depression. Psychotic symptoms have been observed in 14% to 28% of children with 22qDS, but their clinical significance remains uncertain. A 5-year follow-up study of 22qDS children who reported psychotic symptoms at baseline found they had an increased risk for a subsequent psychotic disorder. Thus, a broad differential diagnosis should be considered when 22qDS children present with psychotic symptoms. Longitudinal studies are needed to better understand the full extent of the psychopathology associated with 22qDS. PMID- 19302763 TI - Assessment of depression in children and adolescents. AB - Depression assessment instruments are valuable tools in the treatment of children and adolescents. Available instruments include diagnostic interviews, self administered rating scales, and observer-rated scales. To select an appropriate instrument, the user must define the goal of the assessment and then identify instruments with the properties that match this goal. This article discusses how to choose an assessment instrument and gives an overview of currently available depression assessment instruments. Important considerations include how and by whom an instrument is administered, what kind of data are obtained by the instrument, and the validity and reliability of the instrument. Standardized instruments can greatly improve the assessment process, but the user must not overinterpret or misinterpret the results. PMID- 19302764 TI - Telemedicine in children and adolescents. AB - Psychiatric care for children and adolescents is limited in remote and underserved areas because of the shortage of child and adolescent psychiatrists. Telepsychiatry has the potential to alleviate this problem. This article reviews the procedures used to develop telepsychiatry, equipment needed for videoconferencing in telepsychiatry, benefits and limitations of telepsychiatry, and confidentiality issues in telepsychiatry. Many questions regarding confidentiality, legality, reimbursement, cost-effectiveness, and technology still need to be resolved. However, telepsychiatry has the potential to be a useful treatment alternative. PMID- 19302765 TI - Gender differences in adolescent substance abuse. AB - Gender differences in the epidemiology, comorbidities, and treatment responses of substance abuse have been described in adults. However, a growing body of data suggests that gender differences also exist in adolescents with substance abuse. Unfortunately, research is still limited in this age group. This article reviews gender differences in the diagnosis, presentation, course of illness, and treatment response of substance abuse in adults and adolescents. Adolescent substance abuse treatments that take into account these gender differences are also discussed. PMID- 19302766 TI - The behavioral organization, temporal characteristics, and diagnostic concomitants of rage outbursts in child psychiatric inpatients. AB - Angry outbursts, sometimes called rages, are a major impetus for the psychiatric hospitalization of children. In hospitals, such outbursts are a management problem and a diagnostic puzzle. Among 130 4- to 12-year-olds successively admitted to a child psychiatry unit, those having in-hospital outbursts were likely to be younger, have been in special education, have had a preadmission history of outbursts, and to have a longer hospital stay. Three subsets of behaviors, coded as they occurred in 109 outbursts, expressed increasing levels of anger; two other subsets expressed increasing levels of distress. Factor structure, temporal organization, and age trends indicated that outbursts are exacerbations of ordinary childhood tantrums. Diagnostically, children with outbursts were more likely to have language difficulty and a trend toward attention-deficit/hyperactivity disorder. Outbursts of children with anxiety diagnoses showed significantly more distress relative to anger. Outbursts were not especially associated with our small sample of bipolar diagnoses. PMID- 19302767 TI - Molecular genetics of autism. AB - The increasing autism incidence estimates are generating strong interest in identifying its salient risk factors. Recognition of the importance of genes in this and other disorders has promulgated the development of valuable research tools. As this review indicates, application of these tools paints a portrait of a disorder that is more complex than anticipated. PMID- 19302768 TI - Advances in genetic studies of attention-deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) is among the most common childhood-onset psychiatric disorders. Although family, twin, and adoption studies demonstrate that ADHD is a highly heritable condition, studies also suggest that genetic architecture is complex, prompting the use of more advanced methodologies such as genome-wide linkage and association studies. Although such studies are theoretically compelling, replication of these results has been inconsistent. Meta-analyses have produced more reliable results, but the associations identified to date account for only a small percentage of the genetic component of ADHD. Approaches such as neuroimaging genetics and epigenetic studies are being explored to probe further the etiologic complexity of this disorder. PMID- 19302769 TI - The genetics of reading disability. AB - Genetic factors contribute substantially to the development of reading disability (RD). Family linkage studies have implicated many chromosomal regions containing RD susceptibility genes, of which putative loci at 1p34-p36 (DYX8), 2p (DYX3), 6p21.3 (DYX2), and 15q21 (DYX1) have been frequently replicated, whereas those at 3p12-q12 (DYX5), 6q13-q16 (DYX4), 11p15 (DYX7), 18p11 (DYX6), and Xq27 (DYX9) have less evidence. Association studies of positional candidate genes have implicated DCDC2 and KIAA0319 in DYX2, as well as C2ORF3 and MRPL19 (DYX3), whereas DYX1C1/EKN1 (DYX1) and ROBO1 (DYX5) were found to be disrupted by rare translocation breakpoints in reading-disabled individuals. Four of the candidate genes (DYX1C1, KIAA0319, DCDC2, and ROBO1) appear to function in neuronal migration and guidance, suggesting the importance of early neurodevelopmental processes in RD. Future studies to help us understand the function of these and other RD candidate genes promise to yield enormous insight into the neurobiologic mechanisms underlying the pathophysiology of this disorder. PMID- 19302770 TI - The genetics of childhood-onset schizophrenia: when madness strikes the prepubescent. AB - Stratification by age at onset has been useful for genetic studies across all of medicine. For the past 20 years, the National Institute of Mental Health has been systematically recruiting patients with onset of schizophrenia before age 13 years. Examination of familial transmission of known candidate risk genes was carried out, and a 10% rate of cytogenetic abnormalities was found. Most recently, high-density, array-based scans for submicroscopic rare copy number variations (CNVs) have suggested that this kind of genetic variation occurs more frequently than expected by chance in childhood-onset schizophrenia (COS) and at a higher rate than observed in adult-onset disorder. Several CNVs and cytogenetic abnormalities associated with COS are also seen in autism and mental retardation. Populations with COS may have more salient genetic influence than adult-onset cases. The relationship of rare CNVs to prepsychotic development is being studied further. PMID- 19302771 TI - The genetics of obsessive-compulsive disorder and Tourette's syndrome: what are the common factors? AB - Genetic discovery in obsessive-compulsive disorder and Tourette's syndrome has made significant progress in the past decade. The two disorders are phenomenologically, epidemiologically, and probably pathophysiologically related; however, as with most neuropsychiatric disorders, gene discovery has been challenging. Genetic epidemiology studies support the existence of susceptibility genes in both disorders, and more extensive genome-wide studies are under way. Gene pathways involving neurotransmitter (serotonin, dopamine, glutamate) and neurodevelopment (synaptic, homeobox) domains have been examined, but more complex genetic mechanisms remain largely unexplored. This review addresses the current state of genetic research in obsessive-compulsive disorder and Tourette's syndrome, emphasizing commonalities between the disorders. Questions on common genetic substrates, the use of endophenotypes, and the utility of genetic data to inform pharmacologic treatment are also addressed. PMID- 19302772 TI - The genetics of depression in childhood and adolescence. AB - This article reviews family, twin, and adoption studies of childhood and adolescent depression. Results from several family and twin studies suggest that an etiologic heterogeneity exists in depression in childhood and adolescence. Twin studies show that genetic influences on depression in young people may be indirect and work via effects on environmental risk exposure (gene-environment correlation) or genetic sensitivity to environmental risks (gene-environment interaction). Recent research on gene-environment interaction has examined the effect of specific functional genetic polymorphisms in conjunction with environmental stressors. Future research needs to work toward identifying which environmental and genetic risk factors are crucial to the development of depression in youth, as well as mechanisms involved in the familial transmission of depression. This will not only improve understanding of the etiology of childhood and adolescent depression but also inform the development of therapeutic and preventive interventions. PMID- 19302773 TI - Attention Deficit Hyperactivity Disorder (ADHD) among Male Primary School Children in Dammam, Saudi Arabia: Prevalence and Associated Factors. AB - Attention Deficit Hyperactivity Disorder (ADHD) is one of the most common mental disorders that develop in children and becomes apparent in the preschool and early school years. Several family and maternal-related factors were shown to be associated with ADHD. The aim of the present study was to determine the prevalence of ADHD and associated family and psychosocial factors among male primary school children in Dammam city, Saudi Arabia. A sample size of 1287 students aged 6-13 years in 67 government and 10 private primary schools was selected by multistage systematic random sampling. Data was collected using two types of questionnaires: the modified Arabic version of the Attention Deficit Disorders Evaluation Scale (ADDES) school version, and Parents' questionnaire to diagnose the three main subtypes of ADHD namely: inattention, hyperactivity impulsivity, and combined ADHD. The majority of the boys were from government schools (83.0%), were of age 6-<9 years (40.5%) and of Saudi nationality (80.7%). The overall prevalence of combined ADHD was 16.4%, with a prevalence of 12.4% for hyperactivity-impulsivity and 16.3% for inattention disorders respectively. The study also revealed a variety of family factors to be significantly associated with the development of ADHD. The prevalence of each subtype of ADHD was higher if the child was the 6th one in the family. The prevalence of hyperactivity impulsivity disorder was significantly higher among children living with single parents than those living with both parents (25.0% vs. 12.2% respectively; r =0.016). Inattention was significantly higher among those who had bottle feeding than breastfeeding (21.2% vs. 14.4% respectively; r=0.03). CONCLUSION AND RECOMMENDATIONS: several approaches directed to the child, family, the primary health care services, the school, and the community should be implemented to reduce the prevalence and incidence of ADHD. PMID- 19302774 TI - Prevalence and Determinants of Prehypertension and Hypertension among Preparatory and Secondary School Teachers in Jeddah. AB - Hypertension (HTN) is most likely the most common disease on Earth. It represents the single greatest preventable cause of death in humans and one of the most important modifiable risk factors for cardiovascular diseases. The purpose of this study was to investigate the prevalence and associated risk factors of HTN and pre-hypertension among preparatory and secondary school teachers in Jeddah, KSA, during the educational year 2006 /2007. METHODS: A cross-sectional study was conducted and a multistage stratified random sampling technique was used. HTN (using JNC VII criteria) was defined as Systolic Blood Pressure (SBP) >/= 140 mmHg and / or Diastolic Blood Pressure (DBP) >/= 90 or concurrent use of antihypertensive agents. RESULTS: The prevalence of HTN & pre-hypertension were 25.2 % & 43.0 %, respectively, among the sample of 1476 teachers. Only 30.4 % of hypertensive teachers were aware of their condition. Linear regression indicated that for every 4 Kg increase of weight, there is increase of one mm Hg of SBP and 0.87 mm Hg of DBP. Logistic Regression illustrated that age >/= 40 years (aOR= 4.31, CI: 2.77, 6.73), Body Mass Index (BMI) >/= 25 (aOR= 3.03, CI: 1.77-5.19), males, and diabetes were significantly associated with HTN. Predictors of pre hypertension were male gender (aOR = 3.22, CI: 2.49- 4.16), age >/= 40, and BMI >/= 25. CONCLUSION: The prevalence of pre-hypertension and HTN was high among teachers in Jeddah & BMI was the strongest modifiable risk factor. Lifestyle modification and implementation of screening programs for obesity, diabetes, pre hypertension and HTN are recommended. PMID- 19302775 TI - Teacher's Knowledge, Attitudes and Management Practices about Diabetes Care in Riyadh's Schools. AB - The objective of this study was to assess diabetes-related knowledge, attitudes and management practices among school teachers in order to determine their diabetes training needs and preparedness to provide adequate care for students with diabetes. A cross sectional descriptive study was carried out among 177 school teachers in Boys and Girls primary and intermediate school compounds in Riyadh City. Data was collected using self-administered questionnaires during the period February-March 2007. The results showed that most of the school teachers had fair diabetes knowledge (78%), and unfavorable attitudes toward taking responsibility of diabetes education and care in schools. Recognizing normal, low and high blood sugar levels was the least known. The most frequent sources of information were booklets, brochures, mass media and own experience. A negative significant relationship was found between knowledge and attitude scores. Only 18.6% of teachers had got good total score of diabetes management practices for their diabetic students. The most frequent practices mentioned were trying to have competency in using glucometer, and allowing students to use restroom as needed. Developing an emergency action plan, and observing diabetic students all the school day were the least mentioned practices. Good diabetes managers were more knowledgeable and more expressing unfavorable attitudes. This study highlighted the need of diabetes education training courses especially designed to school teachers to promote adequate care and management of diabetes emergencies in schools. PMID- 19302776 TI - Developing and validating a tool to assess nurse stress. AB - Nursing is generally perceived as a demanding profession. There is a rising necessity for healthcare administrators to tackle the aspects leading to nurse stress and work burnout as a means towards maintaining a stable and continuous workforce at healthcare institutions.The study aimed at the development of a reliable and valid tool for measuring nursing staff stress and burnout at the University Hospital of King Faisal University, Saudi Arabia. A cross-sectional descriptive study was designed. All registered high nurses working at the hospital were included in the study. The total number accounted for 260 nurses. The study revealed a valid and reliable scale. Such results were indicated through face validity, content validity, and principal component analysis using the varimax rotation for the fifteen statements included in the questionnaire. The PCA explained 56.4% of the variance and concluded 3 main components under which the statements would be categorized. Dimensions were given suitable headings as follows; work aspects, working conditions, and workload. Reliability was assessed and revealed an internal consistency (Cronbach alpha=0.8) and a split-half Spearman-Brown coefficient for unequal length r= 0.79. In conclusion, the study demonstrated a short valid and reliable scale to assess the stressful areas for nurses. The scale is convenient for use by healthcare managers at different medical situations. Further studies are recommended for the use of the tool on representative samples of Saudi nurses. PMID- 19302777 TI - Detection of Heterozygous Carriers of PKU in Egypt: Successful Application of a Simple Biochemical Method. AB - The absence of a convenient, direct enzymatic assay for detecting phenylketonuria (PKU) heterozygotes together with the difficulty of the molecular testing due to the large number of mutations in the phenylalanine hydroxylase (PAH) gene has resulted in continued effort to develop an accurate procedure to discriminate the heterozygous individuals from the homozygous normal population. Aiming to find out a method that is simple and reliable for PKU carrier screening, we compared the biochemical data of 20 known PKU obligate heterozygotes with those of 45 presumed normal homozygous controls. Fasting blood samples from all subjects were analyzed for plasma phenylalanine and tyrosine using an amino-acid analyzer. Micromolar plasma concentrations of phenylalanine and tyrosine in addition to Phe/Tyr and Phe2/Tyr ratios were determined and statistical analysis of the difference between the two groups was done using the student's t test. Mean values for phenylalanine concentrations, Phe/Tyr and Phe2/Tyr ratios were significantly higher in PKU heterozygotes than in control subjects. In addition, ROC curve analysis was performed for the same four biochemical variables. The value for the area under the curve (ROCAUC) was obtained for each parameter with the Phe2/Tyr ratio having an area of 1, which means that it had perfect discrimination. When the ratio Phe/Tyr was plotted against the Phe2/Tyr, all the studied control subjects and none of the PKU carriers fell below the values 1.2 and 80, respectively. Finally, by applying the same graphic plot for 20 at-risk PKU family members asking for premarital carrier testing, nine PKU heterozygotes were detected. The same results were successfully reproduced using the values obtained from the ROC curve analysis, indicating a high degree of accuracy for this screening method. In conclusion, the used biochemical method is simple and reliable and it can be useful in the widespread screening for PKU carriers. PMID- 19302778 TI - Cytogenetic studies of couples with reproductive failure in alexandria, egypt. AB - Chromosome abnormalities (CA) are the major contributors to the genetic causes of reproductive disorders. The aim of this study was to assess the frequency and nature of chromosomal aberrations that contribute to the occurrence of reproductive failure in Alexandria, Egypt. The study included 376 couples referred to the outpatient clinic of the Human Genetics Department, Medical Research Institute, University of Alexandria. Chromosome analysis by GTG banding was performed on cultures of peripheral blood lymphocytes. C-banding, and Silver nucleolar organizer region (Ag-NOR) staining were also done when required. The 376 couples with reproductive failure included in this study were divided into groups having: (1) a normal and/or malformed child (NC/MC), (2) two or more spontaneous abortions (>/=2SAB) (3) intrauterine fetal deaths & stillbirths (IUFD &stillbirths) and (4) infertility. Chromosome abnormality was found in 34 couples (9.04%), 26 females (6.91%) and 8 males (2.13%) partners. The structural chromosome rearrangements were more frequent than numerical chromosome aberrations (6.65 and 1.86% of cases, respectively). Chromosomal breaks were found in 0.53% of cases (2/376). The most common structural abnormality was balanced translocations either Robertsonian or reciprocal. 47.XXY karyotype and low mosaicism in X and Y chromosomes were the most common numerical abnormalities. The current study demonstrated the presence of the chromosome abnormality and its influence on reproductive failure. The identification of chromosomal abnormality as the etiology facilitated the counseling and appropriate management. Karyotype analysis should be an integral part of diagnostic work up in couples with reproductive problems especially those undergoing assisted reproductive procedures. PMID- 19302779 TI - Fragile x-syndrome: clinical and molecular studies. AB - Fragile X-syndrome (FXS) is the most common inherited form of mental disability world-wide. The unique mutation that results in FXS consists of expansion of the CGG trinucleotide repeats (>200) in the 3'untranslated region of the Fragile X mental retardation 1 (FMRI) gene at Xq27.3. The aim of the present study was to evaluate the usefulness of the clinical selection prior to undertaking molecular tests for the diagnosis of patients with FXS. Two hundred males, with unknown cause for their mental retardation were scored on eight Fragile X-features. They were selected from the Human Genetics Clinic, Medical Research Institute, Alexandria University, Egypt. Their ages ranged from 2-20 years. All patients were subjected to clinical genetic examination with special emphasis on Fragile X features. RNA analysis was performed to detect cases with full FMRI gene mutation. Amplification of FMRI gene by RT-PCR revealed that 34 cases (17%) had full mutation (i.e. more than 200 repeats). Nineteen Fra X cases (56%) were diagnosed after the age of 10 years. A wide range of Fra X-features ranging from 2-7 features were detected in FXS cases. Significant clinical features in FXS diagnosis included family history of MR, large prominent ears, hyperactivity and macro-orchidism (in post pubertal males). One hundred and twenty seven (76.5%) Fra X -ve MR males had one or no Fra X-features. The eight scoring list, used in this study, allows exclusion from further RNA testing in 63.5% of the retarded males. Clinical preselection for RNA testing for FXS in MR males is feasible using a simple scoring list. This will make RNA analysis more cost effective and will facilitate early detection and management of patients with FXS and proper genetic counseling of their families. PMID- 19302780 TI - Level of lead and cadmium in infant formulae. AB - The concentration of heavy metals, lead (Pb) and cadmium (Cd), were estimated in formula (powdered milk) and cow milk. Eleven samples of infant formulae which represent most kinds of formulae used for feeding infants from birth up to 2 years were taken from pharmacies. Sixteen samples of cow's milk were collected two times during the year from 8 dairy shops in 4 districts in Alexandria. All samples were digested by trichloroacetic acid extraction method and analyzed using atomic absorption. The mean level of Pb and Cd 0.1431+/-0.0402microg/ml and 0.0018+/-0.0006microg/ml, respectively in cow's milk, while it was 0.1563+/ 0.0624microg/ml and 0.0012+/-0.0011microg/ml, respectively in infant formulae. The Estimated Weekly Intake (EWI) for Pb in cow's milk varied from 2.83 5.74microg/kg BW/week. This intake was lower than Provisional Tolerable Weekly Intake (PTWI) of 25 microg/kg BW/week established by WHO expert group. EWI of Cd in cow's milk was also below the PTWI in all samples. In infant formulae EWI of Pb varied between 8.77-24.08 microg/kg BW/week for newborn, 7.08-18.146microg/kg BW/week for six months old and 6.54-22.32 microg/kg Bw /week for one year old. This intake was also lower than PTWI for all different ages, but the highest intake on body weight basis' which is likely to be near to PTWI, was found in infant formulae 2-a and 11-c. PMID- 19302781 TI - Patient skin preparation for surgery. AB - Skin preparation of the patient begins before arriving to the operating room; its purpose is to reduce the risk of post operative wound infection. The objective of this study was to assess the skin preparation for surgical procedures in 3 different hospitals of the General Organization for Teaching Hospitals & Institutes (GOTHI) and to determine their conformity to the existing protocol and make comparisons between them in their performance. A prospective study using the audit was done for 1 month, nearly for the same period in each of the 3 hospitals (A,B,C) to observe the skin preparation practices considering showering, hair removal and the 5 steps of patient skin preparation on table in the operating room. The global results revealed that the compliance rate in the three hospitals together as regards performing the 5 steps of skin preparation in the operating room was 36.2%. It was 50.6% in hospital A, 43.3% in hospital B and 18.7% in hospital C. As regards showering before surgery, it was 55%, removing hair appropriately (clipping) was 23.7% and removing hair by means was 44.6%. PMID- 19302782 TI - Indicators of the screening program for congenital hypothyroidism in alexandria. AB - The screening of newborn children for metabolic and endocrinological disorders is one of the unsung successes of the health service. This study aimed to evaluate the congenital hypothyroidism (CH) screening program in Alexandria in terms of coverage, effectiveness of detecting and managing the cases. A record review was conducted at the Information and Decision Making Support Center of Alexandria Health Directorate during the period from October 2000 to December 2003 to evaluate the coverage of the program and to detect the number of true positives and false positives used to calculate the positive predictive value of the screening test. A follow- up study was conducted at the pediatric clinic of the Student's Sporting Hospital for all positive cases detected by the (CH) screening program attending the clinic for diagnosis, confirmation and treatment during the year 2002 (30 cases were followed-up) . The percentage of program coverage was 49.4%, 63.3% and 77.6% in the years 2001, 2002 and 2003 respectively. The total positive cases in the three years were 26, 30, and 14 with positive predictive value of 65%, 57%, and 64% respectively. About three-quarters (73.3%) of all positive cases (30 cases) were compliant to therapy. Compliance was on fixed times in more than half of cases (56.7%), and irregular in 40%. There was a positive trend of increase in program coverage and effectiveness in detecting and managing the cases through the three years of the study. PMID- 19302783 TI - The long and the short of it: RNA-directed chromatin asymmetry in mammalian X chromosome inactivation. AB - Mammalian X-chromosome inactivation is controlled by a multilayered silencing pathway involving both short and long non-coding RNAs, which differentially recruit the epigenetic machinery to establish chromatin asymmetries. In response to developmentally regulated small RNAs, dicer, a key effector of RNA interference, locally silences Xist on the active X-chromosome and establishes the heterochromatin conformation along the silent X-chromosome. The 1.6 kb RepA RNA initiates silencing by targeting the PRC2 polycomb complex to the inactive X chromosome. In addition, the nuclear microenvironment is implicated in the initiation and maintenance of X-chromosome asymmetries. Here we review new findings involving these various RNA species in terms of understanding Xist gene regulation and the establishment of X-chromosome inactivation. PMID- 19302784 TI - Molecular shape and prominent role of beta-strand swapping in organization of dUTPase oligomers. AB - Most dUTP pyrophosphatases (dUTPases) are homotrimers with interfaces formed between subunit surfaces, in the central channel, and by C-terminal beta-strand swapping. Analysis of intersubunit interactions reveals an important cohesive role for the C-terminus. This is reflected in the crystal structure of fruitfly dUTPase displaying a dimeric organization in crystals grown in alcohol solution, where only beta-strand swapping interactions between subunits are retained from the usual trimer structure. Mutations of a suggested hinge proline destabilize human and Escherichia coli dUTPases without preventing trimeric organization. Trimer formation was, however, prevented in the human enzyme by truncating the C terminus before the swapping arm. The molecular shape of full-length enzymes in solution reveals the localization and variation in flexibility of N- and C terminal segments. PMID- 19302785 TI - Ab initio protein modelling reveals novel human MIT domains. AB - Database searches can fail to detect all truly homologous sequences, particularly when dealing with short, highly sequence diverse protein families. Here, using microtubule interacting and transport (MIT) domains as an example, we have applied an approach of profile-profile matching followed by ab initio structure modelling to the detection of true homologues in the borderline significant zone of database searches. Novel MIT domains were confidently identified in USP54, containing an apparently inactive ubiquitin carboxyl-terminal hydrolase domain, a katanin-like ATPase KATNAL1, and an uncharacterized protein containing a VPS9 domain. As a proof of principle, we have confirmed the novel MIT annotation for USP54 by in vitro profiling of binding to CHMP proteins. PMID- 19302786 TI - Physical association of the catalytic and helper modules of a family-9 glycoside hydrolase is essential for activity. AB - Clostridium thermocellum cellulase 9I (Cel9I) is a non-cellulosomal tri-modular enzyme, consisting of a family-9 glycoside hydrolase (GH9) catalytic module and two family-3 carbohydrate-binding modules (CBM3c and CBM3b). The presence of CBM3c was previously shown to be essential for activity, however the mechanism by which it functions is unclear. We expressed the three recombinant modules independently in Escherichia coli and examined their interactions. Non-denaturing gel electrophoresis, isothermal titration calorimetry, and affinity purification of the GH9-CBM3c complex revealed a specific non-covalent binding interaction between the GH9 module and CBM3c. Their physical association was shown to recover 60-70% of the intact Cel9I endoglucanase activity. PMID- 19302787 TI - Up-regulation of CAR expression through Elk-1 in HepG2 and SW480 cells by serum starvation stress. AB - Constitutive androstane receptor (CAR) is a transcription factor regulating the expression of several genes related to drug metabolism. CAR expression was elevated in human HepG2 and SW480 cells by serum starvation. From reporter gene assays, mutagenesis, RNA interference, and chromatin immunoprecipitation assays, we identified the serum response element at -142/-139 in the CAR gene transactivated by Elk-1. Whereas treatment with U0126 (ERK inhibitor) enhanced CAR expression, SP600125 (stress-activated protein kinase inhibitor, SAPK) suppressed the phosphorylation of Elk-1 caused by serum-starvation stress and the elevation of CAR mRNA, suggesting that CAR expression may be mediated by phosphorylated Elk-1 via the SAPK signaling pathway. PMID- 19302788 TI - Biophysical characterization of a new SCN5A mutation S1333Y in a SIDS infant linked to long QT syndrome. AB - Various entities and genetic etiologies, including inherited long QT syndrome type 3 (LQT3), contribute to sudden infant death syndrome (SIDS). The goal of our research was to biophysically characterize a new SCN5A mutation (S1333Y) in a SIDS infant. S1333Y channels showed the gain of Na(+) channel function characteristic of LQT3, including a persistent inward Na(+) current and an enhanced window current that was generated by a -8 mV shift in activation and a +7 mV shift in inactivation. The correlation between the biophysical data and arrhythmia susceptibility suggested that the SIDS was secondary to the LQT3 associated S1333Y mutation. PMID- 19302789 TI - Release of extracellular membrane vesicles from microvilli of epithelial cells is enhanced by depleting membrane cholesterol. AB - We previously reported on the occurrence of prominin-1-carrying membrane vesicles that are released into body fluids from microvilli of epithelial cells. This release has been implicated in cell differentiation. Here we have characterized these vesicles released from the differentiated Caco-2 cells. We find that in these vesicles, prominin-1 directly interacts with membrane cholesterol and is associated with a membrane microdomain. The cholesterol depletion using methyl beta-cyclodextrin resulted in a marked increase in their release, and a dramatic change in the microvillar ultrastructure from a tubular shape to a "pearling" state, with multiple membrane constrictions, suggesting a role of membrane cholesterol in vesicle release from microvilli. PMID- 19302790 TI - ATM blocks tunicamycin-induced endoplasmic reticulum stress. AB - Endoplasmic reticulum stress (ER-stress) is associated with ataxia telangiectasia mutated (ATM) gene. We present here conclusive data showing that ATM blocks ER stress induced by tunicamycin or ionizing radiation (IR). X-box protein-1 (XBP-1) splicing, GRP78 expression and caspase-12 activation were increased by tunicamycin or IR in Atm-deficient AT5BIVA fibroblasts. Activation of caspase-12 and caspase-3 by tunicamycin was significantly reduced in cells transfected with wild-type Atm (AT5BIVA/wtATM). Atm knockdown by siRNA, however, noticeably elevated ER-stress and chemosensitivity to tunicamycin. In summary, we present substantial data demonstrating that ATM blocks the ER stress signaling associated with cancer cell proliferation. PMID- 19302791 TI - The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction. AB - We have studied the naturally split alpha subunit of the DNA polymerase III (DnaE) intein from Nostoc punctiforme PCC73102 (Npu) using purified proteins and determined an apparent first-order rate constant of (1.1+/-0.2)x10(-2) s(-1) at 37 degrees C. This represents the highest rate reported for the protein trans splicing reaction so far (t(1/2) of approximately 60s). Furthermore, the reaction was very robust and high-yielding with respect to different extein sequences, temperatures from 6 to 37 degrees C, and the presence of up to 6 M urea. Given these outstanding properties, the Npu DnaE intein appears to be the intein of choice for many applications in protein and cellular chemistry. PMID- 19302792 TI - Matrix metalloproteinase-1 expression induced by IL-1beta requires acid sphingomyelinase. AB - Matrix metalloproteinase-1 (MMP-1) is increased in inflammatory conditions leading to destruction of extracellular matrix. Many inflammatory stimuli activate sphingomyelinases (SMases), which generate ceramide. We aimed to define the relevance and type of SMase responsible for the regulation of MMP-1. Acid sphingomyelinase (ASM)-deficient human fibroblasts failed to phosphorylate extracellular signal-regulated kinase (ERK), or upregulate MMP-1 mRNA and protein expression upon stimulation with interleukin-1 beta (IL-1beta), whereas phosphorylation of p38 mitogen-activated protein kinase and IL-8 production remained unaffected. Transfection of ASM restored MMP-1 production. Addition of exogenous SMase was sufficient to restore activation of ERK and increase MMP-1 mRNA. Inhibition of ASM with imipramine completely abrogated MMP-1 induction. The results suggest that IL-1beta-induced expression of MMP-1 is dependent on ASM. PMID- 19302793 TI - SV channels dominate the vacuolar Ca2+ release during intracellular signaling. AB - Vacuoles have long been suggested to mediate a rise in the cytosolic free Ca(2+) during environmental signal transduction. This study addresses the issue of the control of vacuolar calcium release by some of the known signaling molecules such as IP(3), cADPR, ABA, ATP, cAMP, cGMP, H(2)O(2) and CaM. Over 30 concentrations and/or combinations of these signaling compounds were studied in a series of electrophysiological experiments involving non-invasive ion flux measurements (the MIFE) and patch-clamp techniques. Our results suggest that calcium, calmodulin and nucleotides cause calcium release via SV channels. PMID- 19302794 TI - Robustness and topology of the yeast cell cycle Boolean network. AB - Yeast cell cycle Boolean network was used as a case study of robustness to protein noise. Robustness was interpreted as involving stability of G1 steady state and sequence of gene expression from cell cycle START to stationary G1. A robustness measure to evaluate robustness strength of a network was proposed. Robust putative networks corresponding to the same steady state and sequence of gene expression of wild-type network were sampled. Architecture of wild-type yeast cell cycle network can be revealed by average topology profile of sampled robust putative networks. PMID- 19302795 TI - Macrophage inhibitory cytokine MIC-1 is upregulated by short-wavelength light in cultured normal human dermal fibroblasts. AB - To better understand dermal response to visible light, we used DNA microarray analysis to search genes induced by blue or near-UV light in normal human dermal fibroblasts. Of about 12800 transcripts analyzed, near-UV light most prominently upregulated the transcript level of Mic-1, a gene encoding a TGF-beta superfamily protein. Quantitative RT-PCR and immunoblot analyses revealed that mRNA and protein levels of Mic-1 were upregulated by both short-wavelength light but not by green or red light. These results suggest that the human dermis is a site for macrophage inhibitory cytokine-1 (MIC-1) production and that visible light activates a dermal transcription cascade. Considering the role of MIC-1 in immune regulation and appetite control, photic MIC-1 regulation is of physiological importance. PMID- 19302796 TI - Structural and functional analysis of SoPIP2;1 mutants adds insight into plant aquaporin gating. AB - Plant plasma membrane aquaporins facilitate water flux into and out of plant cells, thus coupling their cellular function to basic aspects of plant physiology. Posttranslational modifications of conserved phosphorylation sites, changes in cytoplasmic pH and the binding of Ca(2+) can regulate water transport activity by gating the plasma membrane aquaporins. A structural mechanism unifying these diverse biochemical signals has emerged for the spinach aquaporin SoPIP2;1, although several questions concerning the opening mechanism remain. Here, we describe the X-ray structures of the S115E and S274E single SoPIP2;1 mutants and the corresponding double mutant. Phosphorylation of these serines is believed to increase water transport activity of SoPIP2;1 by opening the channel. However, all mutants crystallised in a closed conformation, as confirmed by water transport assays, implying that neither substitution fully mimics the phosphorylated state. Nevertheless, a half-turn extension of transmembrane helix 1 occurs upon the substitution of Ser115, which draws the C(alpha) atom of Glu31 10 A away from its wild-type conformation, thereby disrupting the divalent cation binding site involved in the gating mechanism. Mutation of Ser274 disorders the C terminus but no other significant conformational changes are observed. Inspection of the hydrogen-bond interactions within loop D suggested that the phosphorylation of Ser188 may also produce an open channel, and this was supported by an increased water transport activity for the S188E mutant and molecular dynamics simulations. These findings add additional insight into the general mechanism of plant aquaporin gating. PMID- 19302797 TI - Viral membrane protein topology is dictated by multiple determinants in its sequence. AB - The targeting, insertion, and topology of membrane proteins have been extensively studied in both prokaryotes and eukaryotes. However, the mechanisms used by viral membrane proteins to generate the correct topology within cellular membranes are less well understood. Here, the effect of flanking charges and the hydrophobicity of the N-terminal hydrophobic segment on viral membrane protein topogenesis are examined systematically. Experimental data reveal that the classical topological determinants have only a minor effect on the overall topology of p9, a plant viral movement protein. Since only a few individual sequence alterations cause an inversion of p9 topology, its topological stability is robust. This result further indicates that the protein has multiple, and perhaps redundant, structural features that ensure that it always adopts the same topology. These critical topogenic sequences appear to be recognized and acted upon from the initial stages of protein biosynthesis, even before the ribosome ends protein translation. PMID- 19302799 TI - Application of methyl-TROSY NMR to test allosteric models describing effects of nucleotide binding to aspartate transcarbamoylase. AB - Aspartate transcarbamoylase has emerged as a textbook example of an allosteric enzyme whose binding of active-site substrates can be explained on the basis of the classical Monod-Wyman-Changeux (MWC) model of allostery. There is still debate, however, regarding the mode of action of ATP and cytidine triphosphate (CTP)--allosteric effectors that bind at regulatory sites 60 A away from the nearest active site. A large body of data for nucleotide binding is consistent with the MWC model, including a previous NMR study showing a shift in the allosteric equilibrium between R and T states that is predicted by this scheme. The possibility of binding-promoted changes to the structures of the active sites, while not within the framework of the MWC model, cannot be excluded, however. Here, the effects of binding of nucleotides are monitored in a series of (1)H-(13)C methyl transverse relaxation optimized spectroscopy spectra recorded on the 300-kDa aspartate transcarbamoylase holoenzyme in both the absence and the presence of saturating amounts of ATP or CTP. No changes in shifts of methyl probes of the catalytic chains (c-chains) that include the active sites are observed, consistent with a lack of structural changes. In addition, methyl (1)H (13)C residual dipolar couplings are measured that are exquisitely sensitive to methyl axis orientations, and correlations between couplings measured on samples with and without nucleotide show no changes in structure of the c-chains. These results indicate that the mechanism of action of ATP and CTP can be explained fully by the MWC model and that any scheme invoking structural changes of the c chains is not correct. PMID- 19302798 TI - The calcium-dependent and calcium-independent membrane binding of synaptotagmin 1: two modes of C2B binding. AB - The Ca2+-independent membrane interactions of the soluble C2 domains from synaptotagmin 1 (syt1) were characterized using a combination of site-directed spin labeling and vesicle sedimentation. The second C2 domain of syt1, C2B, binds to membranes containing phosphatidylserine and phosphatidylcholine in a Ca2+ independent manner with a lipid partition coefficient of approximately 3.0 x 10(2) M(-1). A soluble fragment containing the first and second C2 domains of syt1, C2A and C2B, has a similar affinity, but C2A alone has no detectable affinity to phosphatidylcholine/phosphatidylserine bilayers in the absence of Ca2+. Although the Ca2+-independent membrane affinity of C2B is modest, it indicates that this domain will never be free in solution within the cell. Site directed spin labeling was used to obtain bilayer depth restraints, and a simulated annealing routine was used to generate a model for the membrane docking of C2B in the absence of Ca2+. In this model, the polybasic strand of C2B forms the membrane binding surface for the domain; however, this face of C2B does not penetrate the bilayer but is localized within the aqueous double layer when C2B is bound. This double-layer location indicates that C2B interacts in a purely electrostatic manner with the bilayer interface. In the presence of Ca2+, the membrane affinity of C2B is increased approximately 20-fold, and the domain rotates so that the Ca2+-binding loops of C2B insert into the bilayer. This Ca2+ triggered conformational change may act as a switch to modulate the accessibility of the polybasic face of C2B and control interactions of syt1 with other components of the fusion machinery. PMID- 19302800 TI - Chromium attenuates hepatic damage in a rat model of chronic cholestasis. AB - AIMS: Oxidative stress is involved in cholestasis-induced hepatic damage. Therefore, antioxidant therapy is a recommended therapeutic strategy. Studies have illustrated that chromium can enhance antioxidative capacity leading to a resolution of oxidative stress. The aim of this study was to assess whether chromium has protective effects against cholestasis-related liver damage. MAIN METHODS: Cholestasis was produced by bile duct ligation (BDL) in male Sprague Dawley rats for 3 weeks. Rats were randomly divided into four groups. Control and BDL groups were subjected to sham and BDL operation, respectively, and were supplemented with placebo for 3 weeks. The BDL-post Cr group was supplemented with chromium chloride for 3 weeks after BDL operation. The BDL-pre Cr group was supplemented with chromium chloride for 6 weeks starting from 3 weeks before BDL operation. KEY FINDINGS: In comparison with the control group, the BDL group showed hepatic damage as evidenced by elevation in serum biochemicals, ductular reaction, and fibrosis. These pathophysiological changes were attenuated in the BDL-Pre Cr and BDL-Post Cr groups. However, there was no significant difference between these two groups. The anti-fibrotic effect of chromium was accompanied by reductions in alpha-smooth muscle actin-positive matrix-producing cells and Smad 2/3 activity critical to the fibrogenic potential of transforming growth factor beta 1 (TGF-beta1). In addition, chromium effectively attenuated BDL-induced hepatic oxidative stress. SIGNIFICANCE: The data indicate that chromium attenuates BDL-induced cholestatic liver injury, bile duct proliferation, and fibrosis. The hepatoprotective effect of chromium is associated with antioxidative potential. PMID- 19302801 TI - Differential behavioral and neurochemical effects of exercise, reboxetine and citalopram with the forced swim test. AB - AIMS: In this study, we investigated whether short-term exercise, known to promote hippocampal brain-derived neurotrophic factor (BDNF) expression, would also enhance activity in the Porsolt forced swim test (FST), a model for assessing antidepressant efficacy. We also wished to determine whether exercise combined with antidepressants would be more effective at modifying behavior in the FST than either intervention alone. In parallel with this, we also expected that these interventions would preserve post-stress levels of BDNF, and that antidepressants designed to selectively enhance noradrenergic or serotonergic neurotransmission (reboxetine or citalopram, respectively) would have differential effects on behavior and BDNF expression. MAIN METHODS: Male Sprague Dawley rats were treated with exercise (voluntary wheel running), reboxetine, citalopram, or the combination of exercise and each antidepressant, for 1 week. At the end of this period, a subset of animals from each treatment group underwent the FST. Post-stress levels of hippocampal BDNF mRNA were then quantified via in situ hybridization. KEY FINDINGS: Our results indicate that while both exercise and antidepressant treatment preserved post-stress levels of hippocampal BDNF mRNA, each intervention led to a unique behavioral profile in the FST. We found that antidepressant treatment increased swimming time in the FST, but that exercise decreased swimming time. While the combination of reboxetine-plus-exercise led to an increase in climbing and diving, citalopram plus-exercise reduced these behaviors. SIGNIFICANCE: It is possible that active behaviors during the FST, though specific to antidepressant medications, may not reflect increased hippocampal BDNF expression or other survival- associated benefits. PMID- 19302802 TI - Treatment with a p38 MAPK inhibitor attenuates cisplatin nephrotoxicity starting after the beginning of renal damage. AB - AIMS: Cisplatin (CP) promotes increased production of reactive oxygen species, which can activate p38 mitogen activated protein kinases (p38 MAPKs) leading to apoptosis and increased expression of proinflammatory mediators that intensify the cytotoxic effects of CP. We investigated the effect of the treatment with SB203580, a p38 MAPKs inhibitor, on oxidative stress, on the oxidation-associated signal, p38 MAPK and on apoptosis in CP-injected rats, starting after the beginning of the renal damage. MAIN METHODS: Rats (n=21) were injected with CP (5 mg/kg, i.p.) and 3 and 4 days after some of them (n=8) were treated with SB203580 (0.5 mg/kg, i.p.). Controls (n=6) received saline (i.p.). Two or five days after saline or CP injections, plasma creatinine, urinary volume, sodium and potassium fractional excretions, blood urea nitrogen and urinary lipid peroxidation were measured. The kidneys were removed for histological, apoptosis, immunohistochemical and Western blot studies. KEY FINDINGS: CP caused abnormalities in kidney functions and structure associated with raised urinary peroxidation levels and higher number of apoptotic cells in the outer medulla. The immunostaining studies showed increased numbers of macrophages/monocytes and p-p38 MAPKs positive cells in the renal outer medulla. The increase of p-p38 MAPKs expression was confirmed by Western blot analysis. All of these alterations were attenuated by treatment with SB203580. SIGNIFICANCE: These data suggest that the beneficial effect of SB203580 on CP-induced renal damage might be related, in part, to the blockade of p38 MAPK activation with reduction of the inflammatory process, oxidative stress and apoptotic cell death. PMID- 19302803 TI - Gene expression analysis of human promyelocytic leukemia HL-60 cell differentiation and cytotoxicity induced by natural and synthetic retinoids. AB - AIMS: This study analyzed gene expression profiles of human promyelocytic leukemia HL-60 cells treated with natural and synthetic retinoids (ATRA, RII and R9158), in an attempt to investigate the structure-function relationship of the retinoids in inducing cell differentiation and cytotoxicity. MAIN METHODS: Flow cytometry was used to determine cell cycle changes in HL-60 cells following treatment (1.0 muM) with natural and synthetic retinoids (ATRA, RII and R9158), and cDNA microarrays were used to monitor the gene expression profiles of HL-60 cells treated with the various retinoids. KEY FINDINGS: Consistent with retinoid induced cell differentiation, treatment with these three retinoids correlated with an increase in the percentage of cells arrested in the G1/G0 phase of the cell cycle. Microarray analysis showed upregulation of known differentiation genes, adhesion molecules, and the oxidase activation pathway following retinoid treatment. Differential expression of several genes was observed in HL-60 cells treated with the three retinoids. For example, tissue remodeling protein genes, ubiquitin genes, and signal transduction genes were highly expressed in ATRA- and R9158-treated HL-60 cells, but remained unchanged in HL-60 cells treated with RII. SIGNIFICANCE: The above findings suggest that the differentiation of HL-60 cells induced by the three retinoids occurs through similar pathways, and that there exists a structure-function relationship regarding retinoids and the induction of cell differentiation and cytotoxicity. PMID- 19302805 TI - Placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells support hematopoietic recovery of X-irradiated human CD34+ cells. AB - AIMS: The potential of human mesenchymal stem cell-like stroma prepared from placental/umbilical cord blood for hematopoietic regeneration by X-irradiated hematopoietic stem cells is herein assessed. MAIN METHODS: Placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells were applied to a regenerative ex vivo expansion of X-irradiated human CD34(+) cells in a serum free liquid culture supplemented with a combination of interleukine-3 plus stem cell factor plus thrombopoietin. KEY FINDINGS: The total number of cells and of lineage-committed myeloid hematopoietic progenitor cells generated in the co culture of both non-irradiated and X-irradiated cells with stromal cells was significantly higher than those in the stroma-free culture. In addition, the number of CD34(+) cells and CD34(+)/CD38(-) cells, immature hematopoietic stem/progenitor cells also increased more than the stroma-free culture. The stromal cells produced various types of cytokines, although there was little difference between the co-cultures of non-irradiated and X-irradiated cells with stromal cells. Furthermore, when X-irradiated cells came in contact with stromal cells for 16 h before cytokine stimulation, a similar degree of hematopoiesis was observed, thus suggesting the critical role of cell-to-cell interaction. SIGNIFICANCE: The present results showed the potential efficacy of human mesenchymal stem cell-like stroma for hematopoietic regeneration from irradiated hematopoietic stem/progenitor cells. PMID- 19302804 TI - Metabolic interactions of AGE inhibitor pyridoxamine and antioxidant alpha-lipoic acid following 22 weeks of treatment in obese Zucker rats. AB - AIMS: The advanced glycation end product inhibitor pyridoxamine (PYR) and the antioxidant alpha-lipoic acid (LA) interact to ameliorate insulin resistance in obese Zucker rats following short-term (6-week) treatment. This study was designed to ascertain whether these unique interactive effects of PYR and LA remain manifest following longer-term (22-week) treatment. MAIN METHODS: Female obese Zucker rats received vehicle (OV), PYR (OP, 60 mg/kg body wt), racemic LA (rac-LA; OM, 92 mg/kg), the R-(+)-enantiomer of LA (R-LA; OR, 92 mg/kg), or combined treatments with PYR and rac-LA (OPM) or PYR and R-LA (OPR), daily for 22 weeks. KEY FINDINGS: Individual and combined treatments with PYR, rac-LA, and R LA significantly (p<0.05) inhibited skeletal muscle protein carbonyls (28-36%), a marker of oxidative damage, and triglyceride levels (21-51%). Plasma free fatty acids were reduced in OM (9%), OR (11%), and OPM (16%), with the greatest decrease (26%) elicited in OPR. HOMA-IR, an index of fasting insulin resistance, was decreased in OP (14%) and OPM (17%) groups, with the greatest inhibition (22%) in OPR. Insulin resistance (glucose-insulin index) was lowered (20%) only in OPR. Insulin-mediated glucose transport in isolated skeletal muscle was improved in OM (34%), OR (33%), OPM (48%) and OPR (31%) groups. SIGNIFICANCE: Important interactions between PYR and LA for improvements in glucose and lipid metabolism in the female obese Zucker rat are manifest following a 22-week treatment regimen, providing further evidence for targeting oxidative stress as a strategy for reducing insulin resistance. PMID- 19302806 TI - Acute toxicity of long-circulating and pH-sensitive liposomes containing cisplatin in mice after intraperitoneal administration. AB - AIMS: The objective of this work was to evaluate the acute toxicity of long circulating and pH-sensitive liposomes containing cisplatin (SpHL-CDDP), after their intraperitoneal administration in male and female mice. MAIN METHODS: After single administration of free CDDP (5,10,and 20 mg/kg) or SpHL-CDDP (7,12,30,45 and 80 mg/kg), the body weight was recorded and the LD(50) was calculated. Blood samples were collected for biochemical and hematological analysis. Kidneys, liver, spleen and bone marrow were removed to histopathological examination. KEY FINDINGS: Mice treated with high doses of free CDDP showed a greater loss of body weight and more delayed recovery time than those treated with SpHL-CDDP. The LD(50) values for SpHL-CDDP treatment for male and female mice groups were 2.7 and 3.2 fold higher, respectively, than that obtained for free CDDP. The red and white blood cells counts and quantification of hemoglobin and hematocrit presented no change upon administration of SpHL-CDDP treatment. Free CDDP treatment, however, did lead to an appearance of mild anemia and a reduction in total white blood cell counts. As regards nephrotoxicity, it was observed that free CDDP treatment caused pronounced alterations in the blood urea and creatinine levels of mice. In contrast, these parameters were slightly altered only after SpHL-CDDP treatment at a dose of 30 mg/kg. Microscopic analysis of kidneys from mice treated with SpHL-CDDP showed no morphological alteration. Concerning hepatotoxicity, no histopathological alteration was observed after both treatments. SIGNIFICANCE: These findings reveal that SpHL-CDDP can eliminate CDDP-induced toxicity and is thus a promising candidate for intraperitoneal chemotherapy. PMID- 19302807 TI - Increased effect of fucoidan on lipoprotein lipase secretion in adipocytes. AB - AIMS: Fucoidan, a sulfated polysaccharide extracted from brown seaweed (F. vesiculosus) is recognized as an effective anticoagulant but its anti-lipidemic potency has not been well defined. We investigated the effect of fucoidan on lipoprotein lipase (LPL) secretion by human adipocytes. MAIN METHODS: LPL mRNA and protein expressions were measured using semi-quantitative RT-PCR, ELISA and immunohistochemistry in cultured adipocytes with or without fucoidan treatment. LPL enzyme activity was determined by a fluorometric assay. KEY FINDINGS: In cultured adipocytes, fucoidan induced LPL secretion in a dose- and time-dependent manner. An initial increase in LPL was maintained at a significant level but much slower than that in heparin-treated cells. Fucoidan also dose-dependently induced a cofactor of LPL, the apolipoprotein C-II (ApoC-II) secretion. In fucoidan treated cells, LPL mRNA was time-dependently increased and LPL protein expression was also inceased. Treatment with both heparin and fucoidan showed no further increase in media LPL activity compared to heparin alone. In the conditioned medium from fucoidan-treated cells followed for 4 h, LPL activity decayed exponentially with half-life of about 180 min. In addition, the extracellular LPL mass in cycloheximide (a protein synthesis inhibitor) and fucoidan-treated cells did not change markedly, but LPL shifted significantly from active to inactive form. SIGNIFICANCE: These results suggest that fucoidan acts like heparin by releasing LPL in addition to increasing the intracellular transport and decreasing the degradation of LPL in the medium. Furthermore, LPL and ApoC-II secretion induced by fucoidan may be involved in regulating plasma triglyceride lowering clearance. PMID- 19302808 TI - Procognitive 5-HT6 antagonists in the rat forced swimming test: potential therapeutic utility in mood disorders associated with Alzheimer's disease. AB - AIMS: 5-HT(6) receptor subtype is predominantly expressed in the brain, and preclinical evidence suggests its potential role in the cognitive function. Brain microdialysis studies demonstrated that 5-HT(6) antagonists enhance not only cholinergic but also monoaminergic neurotransmission, a property that may differentiate from acetylcholine esterase (AChE) inhibitors such as donepezil. In this study we compared the antidepressant-like effects of 5-HT(6) antagonists with donepezil to determine whether their different effects on monoamines are behaviorally relevant. MAIN METHODS: Selective 5-HT(6) antagonists (SB-399885 and SB-271046) and donepezil were evaluated in the rat forced swimming test since this is known to identify drugs such as antidepressants which can increase brain monoamine levels. Binding assay was undertaken by using [(125)I]SB-258585 to measure brain 5-HT(6) receptor occupancy. KEY FINDINGS: Systemic administration of SB-399885 (3 and 10 mg/kg, i.p.) and SB-271046 (10 and 30 mg/kg, i.p.) produced a significant reduction of immobility time in the rat forced swimming test with a similar profile in terms of 5-HT(6) receptor occupancy (62 and 96% for 3 and 10 mg/kg SB-399885 respectively; 56 and 84% for 10 and 30 mg/kg SB 271046 respectively). In contrast, donepezil (0.5 and 1 mg/kg i.p.) did not show any effects in this model. SIGNIFICANCE: These data suggest that 5-HT(6) antagonists, at doses corresponding to those occupy central 5-HT(6) receptors, could have an antidepressive effect in humans. This may differentiate 5-HT(6) antagonists from AChE inhibitors with respect to the mood control in the symptomatic treatment of Alzheimer's disease. PMID- 19302809 TI - Pre-ischemic treadmill training affects glutamate and gamma aminobutyric acid levels in the striatal dialysate of a rat model of cerebral ischemia. AB - AIMS: Treadmill training has been shown to improve function in animal models and patients with cerebral ischemia. However, the neurochemical effects of this intervention on the ischemic brain have not been well studied. This study was designed to evaluate the effects of pre-ischemic treadmill training on the release of glutamate and gamma-aminobutyric acid (GABA) from the striatum in a rat middle cerebral artery occlusion (MCAO) model. MAIN METHODS: Rats were divided into five groups: sham control without MCAO, and 0, 1, 2 and 4 weeks pre ischemic treadmill training. After training, cerebral ischemia was induced by MCAO for 120 min, followed by reperfusion. Microdialysis was used to collect dialysates from the striatum immediately before ischemia, and at 40, 80 and 120 min after ischemia, as well as at 40, 80, 120, 160, 200 and 240 min after reperfusion. KEY FINDINGS: Pre-ischemic treadmill training decreased glutamate release and increased GABA release during the acute phase of cerebral ischemia/reperfusion. Treadmill training for at least 2 weeks produced statistically significant changes in GABA/glutamate release. SIGNIFICANCE: The present study suggests that treadmill training inhibits the excessive release of glutamate, by stimulating GABA release during the acute phase of cerebral ischemia. This may be one of the important mechanisms to protect the striatal neurons from ischemic damage. PMID- 19302810 TI - Specific induction of a 72-kDa heat shock protein protects esophageal mucosa from reflux esophagitis. AB - AIMS: The aim of this study is to investigate the expression and cytoprotective function of a 72-kDa heat shock protein (HSP72) using a reflux esophagitis model in rats. MAIN METHODS: Expression of HSP60, HSP72, and HSP90 in rat esophageal mucosa was evaluated by Western blot analysis before and after hyperthermia (42.5 degrees C, 20 min). Rats received the operation to produce reflux esophagitis with or without pretreatment with hyperthermia to induce HSPs. The esophageal mucosal damage was evaluated 12 h after the operation. KEY FINDINGS: Expression of HSP72 was significantly increased by hyperthermia in rat esophageal mucosa. Reflux esophagitis was dramatically prevented when HSP72 was preinduced by hyperthermia. Furthermore, activation of TNF-alpha and IL-1beta in esophageal mucosa was also suppressed. SIGNIFICANCE: These results suggested that hyperthermia protects the esophageal mucosa in reflux esophagitis model by inducing HSP72 and suppressing proinflammatory cytokine activation. These findings might suggest that HSP-inducing therapy could be a novel and unique therapy for reflux esophagitis. PMID- 19302811 TI - Apoptotic signaling induced by H2O2-mediated oxidative stress in differentiated C2C12 myotubes. AB - AIMS: Apoptotic signaling proteins were evaluated in postmitotic skeletal myotubes to test the hypothesis that oxidative stress induced by H(2)O(2) activates both caspase-dependent and caspase-independent apoptotic proteins in differentiated C2C12 myotubes. We hypothesized that oxidative stress would decrease anti-apoptotic protein levels in C2C12 myotubes. MAIN METHODS: Apoptotic regulatory factors and apoptosis-associated proteins including Bcl-2, Bax, Apaf 1, XIAP, ARC, cleaved PARP, p53, p21(Cip1/Waf1), c-Myc, HSP70, CuZnSOD, and MnSOD protein content were measured by immunoblots. KEY FINDINGS: H(2)O(2) induced apoptosis in myotubes as shown by DNA laddering and an elevation of apoptotic DNA fragmentation. Cell death ELISA showed increase in the extent of apoptotic DNA fragmentation following treatment with H(2)O(2). Treatment with 4 mM of H(2)O(2) for 24 or 96 h caused increase in Bax (56%, 227%), cytochrome c (282%, 701%), Smac/DIABLO (155%, 260%), caspase-3 protease activity (51%, 141%), and nuclear and cytosolic p53 (719%, 1581%) levels in the myotubes. As an estimate of the mitochondrial AIF release to the cytosol, AIF protein content measured in the mitochondria-free cytosolic fraction was elevated by 65% after 96 h treatment with 4 mM of H(2)O(2). AIF measured in the nuclear protein fraction increased by 74% and 352% following treatment with 4 mM of H(2)O(2) for 24 and 96 h, respectively. Bcl-2 declined in myotubes by 61% and 69% after 24 or 96 h of treatment in 4 mM H(2)O(2), respectively. SIGNIFICANCE: These findings indicate that both caspase-dependent and caspase-independent mechanisms are involved in coordinating the activation of apoptosis induced by H(2)O(2) in differentiated myotubes. PMID- 19302812 TI - TNF-alpha and IL-1beta inhibit RUNX2 and collagen expression but increase alkaline phosphatase activity and mineralization in human mesenchymal stem cells. AB - AIMS: Joint inflammation leads to bone erosion in rheumatoid arthritis (RA), whereas it induces new bone formation in spondyloarthropathies (SpAs). Our aims were to clarify the effects of tumour necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta) on osteoblast differentiation and mineralization in human mesenchymal stem cells (MSCs). MAIN METHODS: In MSCs, expression of osteoblast markers was assessed by real-time PCR and ELISA. Activity of tissue nonspecific alkaline phosphatase (TNAP) and mineralization were determined by the method of Lowry and alizarin red staining respectively. Involvement of RUNX2 in cytokine effects was investigated in osteoblast-like cells transfected with a dominant negative construct. KEY FINDINGS: TNF-alpha (from 0.1 to 10 ng/ml) and IL-1beta (from 0.1 to 1 ng/ml) stimulated TNAP activity and mineralization in MSCs. Addition of 50 ng/ml of IL-1 receptor antagonist in TNF-alpha-treated cultures did not reverse TNF-alpha effects, indicating that IL-1 was not involved in TNF-alpha-stimulated TNAP activity. Both TNF-alpha and IL-1beta decreased RUNX2 expression and osteocalcin secretion, suggesting that RUNX2 was not involved in mineralization. This hypothesis was confirmed in osteoblast-like cells expressing a dominant negative RUNX2, in which TNAP expression and activity were not reduced. Finally, since mineralization may merely rely on increased TNAP activity in a collagen-rich tissue, we investigated cytokine effects on collagen expression, and observed that cytokines decreased collagen expression in osteoblasts from MSC cultures. SIGNIFICANCE: The different effects of cytokines on TNAP activity and collagen expression may therefore help explain why inflammation decreases bone formation in RA whereas it induces ectopic ossification from collagen-rich entheses during SpAs. PMID- 19302813 TI - Subchronic stress-induced depressive behavior in ovariectomized mice. AB - AIMS: Mood disorders including depression are more common in women than men, particularly in times of lower estradiol levels. In this study, we investigated the effect of estrogen on emotional behavior in mice in a stress environment. MAIN METHODS: Female mice were divided into four groups: two groups were ovariectomized (OVX) and two were sham-operated. One group each of OVX and sham mice was kept in a normal environment and the other groups were assigned to a daily stress (1 h/day) for 7 days from 5 days after operation. On the 14th day after operation, subjects were measured to assess behavioral specificity, locomotor activity, elevated plus-maze (EPM) behavior, passive avoidance (PA) behavior and forced swimming behavior. KEY FINDINGS: The OVX plus stress (OVX+S) group showed a significant prolongation of immobility compared with the other groups. In all the groups there were no changes in locomotor activity, EPM behavior or PA behavior. We further examined the effect of estrogen against depressive behavior in the OVX+S group. The vehicle or 17beta-estradiol (E2) was administered s.c. to OVX+S mice for 4 days beginning on post-operative day 11. Subchronic E2 treatment decreased the stress response and improved depressive behavior relative to the vehicle group. SIGNIFICANCE: These data have important implications regarding the prevention of depression in postmenopausal women undergoing estrogen therapy. PMID- 19302814 TI - Monohydroxylated polycyclic aromatic hydrocarbons inhibit both osteoclastic and osteoblastic activities in teleost scales. AB - AIMS: We previously demonstrated that monohydroxylated polycyclic aromatic hydrocarbons (OHPAHs) bound to a human estrogen receptor (ER) by a yeast two hybrid assay, but polycyclic aromatic hydrocarbons did not have a binding activity. Therefore, the direct effect of 3-hydroxybenz[a]anthracene (3-OHBaA) and 4-hydroxybenz[a]anthracene (4-OHBaA) on osteoclasts and osteoblasts in teleosts was examined. As a negative control, 1-hydroxypyrene (1-OHPy), which has no binding activity to human ER, was used. MAIN METHODS: The effect of OHPAHs on osteoclasts and osteoblasts was examined by an assay system using teleost scale as each marker: tartrate-resistant acid phosphatase for osteoclasts and alkaline phosphatase for osteoblasts. Changes in cathepsin K (an osteoclastic marker) and insulin-like growth factor-I (IGF-I) (an osteoblastic marker) mRNA expressions in 4-OHBaA-treated goldfish scales were examined by using a reverse transcription polymerase chain reaction. KEY FINDINGS: In both goldfish (a freshwater teleost) and wrasse (a marine teleost), the osteoclastic activity in the scales was significantly suppressed by 3-OHBaA and 4-OHBaA, although 1-OHPy did not affect the osteoclastic activity. In reference to osteoblasts, the osteoblastic activity decreased with both 3-OHBaA and 4-OHBaA and did not change with the 1-OHPy treatment. However, 17beta-estradiol (E(2)) significantly increased both the osteoclastic and osteoblastic activities in the scales of both goldfish and wrasse. The mRNA expressions of both cathepsin K and IGF-I decreased in the 4 OHBaA-treated scales but increased in the E(2)-treated scales. SIGNIFICANCE: The current data are the first to demonstrate that 3-OHBaA and 4-OHBaA inhibited both osteoclasts and osteoblasts and disrupted the bone metabolism in teleosts. PMID- 19302815 TI - The diagonal band of Broca is involved in the pressor pathway activated by noradrenaline microinjected into the periaqueductal gray area of rats. AB - AIMS: The dorsal periaqueductal gray area (dPAG) is involved in cardiovascular modulation. Previously, we reported that noradrenaline (NA) microinjection into the dPAG caused a pressor response that was mediated by vasopressin release into the circulation. However, the neuronal pathway that mediates this response is as yet unknown. There is evidence that chemical stimulation of the diagonal band of Broca (dbB) also causes a pressor response mediated by systemic vasopressin release. In the present study, we evaluated the participation of the dbB in the pressor response caused by NA microinjection into the dPAG as well as the existence of neural connections between these areas. MAIN METHODS: With the above goal, we verified the effect of the pharmacological ablation of the dbB on the cardiovascular response to NA microinjection into the dPAG of unanesthetized rats. In addition, we microinjected the neuronal tracer biotinylated-dextran amine (BDA) into the dPAG and looked for efferent projections from the dPAG to the dbB. KEY FINDINGS: The pharmacologically reversible ablation of the dbB with local microinjection of CoCl(2) significantly reduced the pressor response caused by NA microinjection (15 nmol/50 nL) into the dPAG. In addition, BDA microinjection into the dPAG labeled axons in the dbB, pointing to the existence of direct connections between these areas. SIGNIFICANCE: The present results indicate that synapses within the dbB are involved in the pressor pathway activated by NA microinjection into the dPAG and direct neural projection from the dPAG to the dbB may constitute the neuroanatomic substrate for this pressor pathway. PMID- 19302816 TI - Nm23-H1 regulates the proliferation and differentiation of the human chronic myeloid leukemia K562 cell line: a functional proteomics study. AB - AIMS: Nm23-H1 is a suppressor of metastasis that has been implicated in the regulation of proliferation and differentiation of hematopoietic cells, although specific mechanisms for Nm23-H1 have not been well-characterized. Our study is designed to further elucidate the role of Nm23-H1 in the human chronic myeloid leukemia K562 cell line. MAIN METHODS: In this study we generated and selected two cell clone pools of human chronic myeloid leukemia K562 cells with up regulated and down-regulated Nm23-H1 expression. KEY FINDINGS: Our data show that knockdown of Nm23-H1 decreased proliferation and increased the percentage of cells arrested in the G0/G1 phase of the cell cycle. Correspondingly, K562 cells overexpressing Nm23-H1 were more proliferative. After treatment of these two cell types with phorbol 12-myristate 13-acetate (PMA) for 48 h, cells with reduced Nm23-H1 expression had a higher percentage of 8N ploidy and higher expression of CD41 than K562 cells overexpressing Nm23-H1. A functional proteomics analysis identified ten proteins, including ANP32A, Cdc42GAP, and the isoform 2 of SET, whose expression levels were significantly altered by down-regulation of Nm23-H1. In addition, cells with decreased levels of Nm23-H1 had significantly reduced expression of Cdc42 independent of treatment with PMA. The interaction of the endogenous Nm23-H1 and Cdc42 proteins has been further validated by reciprocal immunoprecipitations. SIGNIFICANCE: We provide data that complement functional studies of Nm23-H1 in regulating hematopoietic cells, and address action mechanisms of Nm23-H1 that have not previously been reported. PMID- 19302817 TI - TNFalpha- and NF-kappaB-dependent induction of the chemokine CCL1 in human macrophages exposed to the atherogenic lipoprotein(a). AB - AIMS: CCL1 is a chemokine thought to contribute to cardiovascular diseases and recently reported to be regulated by the pro-atherogenic lipoprotein(a) (Lp(a)) and the ligand-activated aryl hydrocarbon receptor (AhR). The present study was designed to investigate molecular regulatory pathways involved in Lp(a)-mediated induction of CCL1. MAIN METHODS: CCL1 regulation was studied in Lp(a)-exposed human primary macrophages using mainly quantitative reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay and electrophoretic mobility shift assay (EMSA). KEY FINDINGS: Using the AhR antagonist alpha napthtoflavone, the translational inhibitor cycloheximide and anti-tumor necrosis factor alpha (TNFalpha) neutralizing antibodies, we demonstrated that Lp(a) mediated mRNA induction of CCL1 occurs in an AhR-independent manner and requires de novo protein synthesis of TNFalpha. Involvement of this cytokine was further underlined by the fact that it increased expression and secretion of CCL1 by itself in macrophages. DNA binding activity of NF-kappaB, a well-known molecular effector of TNFalpha, was moreover activated by Lp(a) in a TNFalpha-dependent manner and the use of the NF-kappaB inhibitor Bay 11-7082 blocked Lp(a)-triggered CCL1 induction. In addition, Lp(a) induced binding of NF-kappaB to a NF-kappaB consensus element on CCL1 promoter as assessed by EMSA. Co-exposure to Lp(a) and the AhR ligand benzo(a)pyrene was finally shown to superinduce CCL1 expression in human macrophages, supporting the conclusion that Lp(a) and AhR ligands act on CCL1 through independent ways. SIGNIFICANCE: These data suggest that Lp(a) triggered induction of CCL1 expression is mediated by TNFalpha and subsequent activation of NF-kappaB, without AhR involvement. PMID- 19302818 TI - Evidence of O-linked N-acetylglucosamine in diabetic nephropathy. AB - AIMS: There is increasing evidence that O-linked N-acetylglucosamine (O-GlcNAc) plays an important role in cell signaling pathways. It has also been reported that increases in O-GlcNAc contribute to the development of diabetes and diabetic complications; however, little is known about O-GlcNAc levels in diabetic nephropathy (DNP). Therefore the goal of this study was to determine whether O GlcNAc could be detected in human kidney biopsy specimens, and if so to examine whether O-GlcNAc levels were increased in the kidneys of patients with DNP compared to the non-diabetic individuals. MAIN METHODS: Kidney biopsy specimens were obtained from type-2 diabetic patients (n=6) and patients diagnosed with thin basement membrane nephropathy (n=7) were used as non-diabetic controls. O GlcNAc levels were assessed by immunohistochemistry using the anti-O-GlcNAc antibody CTD110.6. KEY FINDINGS: We show that O-GlcNAc modification of proteins can be detected in the human kidney biopsy specimens. Furthermore, in diabetic patients, we found significantly increased numbers of O-GlcNAc positive cells in the glomeruli and significantly elevated staining in the tubuli (both in the nucleus and in the cytosol). In addition we also observed an intense, granular O GlcNAc staining specifically in diabetic tubuli. SIGNIFICANCE: In light of the increase in O-GlcNAc staining in the diabetic patients, we propose that increased O-GlcNAc levels might contribute to the development of diabetic nephropathy. PMID- 19302819 TI - Effect of preischemic beta-adrenoceptor stimulation on postischemic contractile dysfunction. AB - AIMS: Short periods of preischemic beta-adrenoceptor stimulation protect hearts against postischemic left ventricular dysfunction. It was the aim of this study to decide whether this procedure mimics ischemic preconditioning by the generation of preischemic hemodynamic and energetic stress or whether it represents an endogenous phenomenon and to investigate the influence of age and hypertension. MAIN METHODS: Isolated rat hearts were investigated ex vivo by Langendorff perfusion and exposed to an established ischemia/reperfusion protocol (45 min no-flow ischemia and 90 min reperfusion). Left ventricular developed pressure (LVDP), rate pressure product, and +/-dP/dt were analyzed. KEY FINDINGS: Isoprenaline concentration dependently increased LVDP up to 40+/-15 mm Hg (approximately EC(50) of 9.9+/-0.5 nM). Isoprenaline given prior to ischemia attenuated the subsequent postischemic ventricular dysfunction (approximately EC(50) of 1.4+/-0.2 pM). However, concentrations high enough to improve LVDP in normoxic hearts did not improve postischemic recovery albeit a significant reduction of hypercontraction-induced cell damage. The effect on functional recovery was attenuated by atenolol, H89, and wortmannin suggesting that beta adrenoceptor stimulation, protein kinase A, and PI 3-kinase activation are involved. The effect was conserved in hearts from 13 month old rats but lost in age-matched spontaneously hypertensive rats. SIGNIFICANCE: The study identifies preischemic beta-adrenoceptor stimulation as a pharmacological preconditioning protocol that does not simply mimic classical ischemic preconditioning by induction of hemodynamic or energetic stress prior to a prolonged ischemic period. The observed loss of effectiveness in hypertensives may contribute to the reduced ischemic tolerance of hypertensives. PMID- 19302820 TI - Pharmacological preconditioning with erythropoietin reduces ischemia-reperfusion injury in the small intestine of rats. AB - AIMS: Considering the implications that arose from several recent experimental studies using recombinant human erythropoietin in rodents, erythropoietin has been regarded as a pharmacological preconditioning agent. The purpose of the present study was to evaluate whether erythropoietin has a preconditioning effect against ischemia and reperfusion injury in the small intestine of the rat. MAIN METHODS: Intestinal ischemia was induced in male Wistar rats by clamping the superior mesenteric artery for 30 min, followed by reperfusion for 180 min. Recombinant human erythropoietin (1000 or 3000 U/kg) or vehicle was administered intraperitoneally 24 h prior to ischemia. After collection of ileal tissue, evaluation of damage was based on measurements of the accumulation of polymorphonuclear neutrophils by technetium-99m-labeled leukocyte uptake, content of malondialdehyde, reduced glutathione, contractile responses to agonists, and an evaluation of histopathological features in intestinal tissue. KEY FINDINGS: Treatment with erythropoietin 24 h before ischemia significantly reduced the tissue content of malondialdehyde and increased that of reduced glutathione. Pretreatment also significantly suppressed leukocyte infiltration into the postischemic tissue, as evidenced by the lower content of myeloperoxidase and technetium-99m-labeled leukocytes. Physiological and histopathological improvements were also significant with the rHuEpo treatment. SIGNIFICANCE: Results of the present study indicate that rHuEpo is an effective preconditioning agent in ischemic injury of the small intestine. Protection provided by recombinant human erythropoietin is closely related to the inhibition of oxidative stress and leukocyte infiltration, which might be among the possible protective mechanisms of erythropoietin in intestinal ischemia and reperfusion. PMID- 19302821 TI - Antioxidative effect of the herbal remedy Qin Huo Yi Hao and its active component tetramethylpyrazine on high glucose-treated endothelial cells. AB - AIMS: This study was designed to gain insights into the antioxidant mechanism of a Chinese herbal remedy, Qing Huo Yi Hao (QHYH), and its active components against oxidative stress induced by high glucose in endothelial cells. MAIN METHODS: Effects of QHYH on reactive oxygen species (ROS) production and nitric oxide (NO) generation were measured with the fluorescent markers H(2)DCF-DA and DAF-FM DA, respectively. Phosphorylation of Akt (protein kinase B)/eNOS (endothelial nitric oxide synthase) and uncoupling protein 2 (UCP2) expression were studied by Western blot techniques. Influences of QHYH and one of the active components (tetramethylpyrazine, TMP) on UCP2 expression were subsequently evaluated by quantitative real-time reverse transcription-polymerase chain reaction. Using RNA interference techniques, the involvement of UCP2 in high glucose-induced ROS production in mouse brain microvascular (bEnd.3) cells and its correlation with the antioxidant effect of QHYH were further assessed. KEY FINDINGS: Our results showed that QHYH could protect endothelial cells from high glucose-induced damages, such as ROS production, down-regulation of Akt/eNOS phosphorylation and reduction of NO generation. The protective properties of QHYH were partially attributed to UCP2 mRNA/protein expression, because silence of UCP2 gene by siRNAs (small interfering RNAs) abolished such effects. A total of 28 extracts and 11 active components isolated from QHYH were functionally analyzed. Of which, TMP displayed comparable antioxidant and endothelial protective effects as QHYH. SIGNIFICANCE: All of the data, taken together, point to some therapeutic potential of QHYH and TMP for vascular complications of diabetes. PMID- 19302822 TI - Involvement of Sp1 binding sequences in basal transcription of the rat fibroblast growth factor-2 gene in neonatal cardiomyocytes. AB - AIMS: The identification of transcription factors that regulate the transcription of the fibroblast growth factor-2 (FGF-2) gene has facilitated the understanding of the etiology of cardiovascular diseases. The purpose of this study was to determine the molecular mechanism underlying the activation of FGF-2 gene transcription in cardiomyocytes from neonatal rats. MAIN METHODS: To identify the factors involved in cardiac expression of FGF-2, we used transient transfections in neonatal rat cardiomyocytes coupled with electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) analyses. KEY FINDINGS: Deletion analyses showed that the region between -16 and +59 was essential for maximal FGF 2 promoter activity. Three putative stimulating protein 1 (Sp1) regulatory sites located at positions -3, +14, and +27 were predicted within this region by computer analysis. EMSA showed the existence of two atypical G-rich Sp1-binding elements located at positions -3 and +14. Mutation of these two sites resulted in a significant decline in FGF-2 promoter activity compared to wild type promoter activity. Combinatorial mutation of these sites reduced the promoter activity to background levels. Mutation of the Sp1 motif at +27 did not affect promoter activity. Lastly, ChIP analyses revealed that Sp1 binds to the FGF-2 promoter region in vivo. SIGNIFICANCE: These results indicate that expression of FGF-2 in neonatal rat cardiomyocytes is associated with Sp1 binding to the FGF-2 promoter. PMID- 19302823 TI - Modulation of proliferation and differentiation of C2C12 skeletal muscle cells by fatty acids. AB - AIMS: This study was performed to elucidate whether mitogen-activated protein kinases (MAPKs) are involved in the modulation of the proliferation and differentiation of skeletal muscle cells by fatty acids. MAIN METHODS: C2C12 myoblasts were cultured in differentiation medium containing 2% horse serum for 3 days, and treated with each fatty acid. Phosphorylation levels of MAPKs were examined by immunoblot analysis. KEY FINDINGS: The mono-unsaturated fatty acids (MUFAs), oleic acid (OA) and n-6 polyunsaturated fatty acids (n-6 PUFAs), linoleic acid (LA), gamma-linoleic acid (GLA), and arachidonic acid (AA) increased the proliferation of C2C12 cells. On the other hand, n-3 polyunsaturated fatty acids (n-3 PUFAs) and saturated fatty acids (SFs) did not affect the proliferation of C2C12 cells. In addition, the treatment of cis-9, trans-11 conjugated linoleic acid (c9,t11 CLA) showed an increased cell proliferation. However, trans-10, cis-12 conjugated linoleic acid (t10,c12 CLA) significantly inhibited cell proliferation. Treatment of C2C12 cells with LA, OA, and c9,t11 CLA increased phosphorylation levels of ERK1/2 and JNK during proliferation. During cell differentiation, OA, LA, and c9,t11 CLA stimulated differentiation of C2C12 cells, whereas t10,c12 CLA inhibited differentiation. We also found that OA, LA, and c9, t11 CLA increased phosphorylation level of ERK1/2, but not JNK during differentiation. SIGNIFICANCE: These results suggest that fatty acids are able to modulate the proliferation and differentiation of skeletal muscle and MAPKs may be involved in the modulation of the proliferation and differentiation of skeletal muscle cells by fatty acids. PMID- 19302825 TI - Aberrant intracellular IGF-1R beta-subunit makes receptor knockout cells (IGF1R-/ ) susceptible to oncogenic transformation. AB - Insulin-like growth factor 1 receptor (IGF-1R) is important for transformation of cells with cellular and viral oncogenes. This knowledge is mainly based on experiments on IGF-1R knockout mouse fibroblasts, which mostly are unable to transform after introduction of various oncogenes. Recently, we observed two variants of R- cells, one of which (R-s) surprisingly expresses the beta-subunit of IGF-1R whereas the other one (R-r) does not. Here we show that the beta subunit is localized intracellularly and forms perinuclear aggregates. It expresses tyrosine kinase activity and appears to be crucial for cell survival since knockdown of it kills the R-s cells. H-RasV12 and/or polyoma middle T antigen fail to transform R-r, whereas R- cells expressing the beta-subunit were transformed as assessed by formation of colonies in soft agar. The oncogenic transformation of R-s cells was, however, abrogated when the aberrant beta subunit was knockdown by siRNA. The occurrence of intracellular IGF-1R, especially in tumor cells, has been widely reported but its function has not been understood. Our study provides evidence that it may be important for cell survival and transformation. PMID- 19302826 TI - Organizing action of prenatally administered testosterone propionate on the tissues mediating mating behavior in the female guinea pig. AB - Our "Organizing Action" paper published in 1959 put forward the concept that prenatal exposure to testosterone masculinized the behavior of genetic female guinea pigs. Specifically, we proposed that testosterone or some metabolite acted on the central nervous tissues in which patterns of sexual behavior are organized. We later went on to demonstrate similar effects in rhesus monkeys by showing that play behavior by female monkeys prenatally treated with testosterone was masculinized as well. These findings support the organizing actions of androgens as a general process of sexual differentiation. PMID- 19302827 TI - Novel signaling pathways promote a paracrine wave of prostacyclin-induced vascular smooth muscle differentiation. AB - The important athero-protective role of prostacyclin is becoming increasingly evident as recent studies have revealed adverse cardiovascular effects in mice lacking the prostacyclin receptor, in patients taking selective COX-2 inhibitors, and in patients in the presence of a dysfunctional prostacyclin receptor genetic variant. We have recently reported that this protective mechanism includes the promotion of a quiescent differentiated phenotype in human vascular smooth muscle cells (VSMC). Herein, we address the intriguing question of how localized endothelial release of the very unstable eicosanoid, prostacyclin, exerts a profound effect on the vascular media, often 30 cell layers thick. We report a novel PKA-, Akt-1- and ERK1/2-dependent prostacyclin-induced prostacyclin release that appears to play an important role in propagation of the quiescent, differentiated phenotype through adjacent arterial smooth muscle cells in the vascular media. Treating VSMC with the prostacyclin analog iloprost induced differentiation (contractile protein expression and contractile morphology), and also up-regulated COX-2 expression, leading to prostacyclin release by VSMC. This paracrine prostacyclin release, in turn, promoted differentiation and COX-2 induction in neighboring VSMC that were not exposed to iloprost. Using siRNA and pharmacologic inhibitors, we report that this positive feedback mechanism, prostacyclin-induced prostacyclin release, is mediated by cAMP/PKA signaling, ERK1/2 activation, and a novel prostacyclin receptor signaling pathway, inhibition of Akt-1. Furthermore, these pathways appear to be regulated by the prostacyclin receptor independently of one another. We conclude that prevention of de-differentiation and proliferation through a paracrine positive feedback mechanism is a major cardioprotective function of prostacyclin. PMID- 19302828 TI - Antidepressant-like effects of curcumin on serotonergic receptor-coupled AC-cAMP pathway in chronic unpredictable mild stress of rats. AB - Serotonergic receptors take their physiologic effects by affecting adenylyl cyclase (AC) catalytic activity and cyclic adenosine monophosphate (cAMP) concentration. AC-cAMP second messenger pathway has been recently suggested to play an important role in depression. Therefore, the compound that regulates the signal pathway may have potential as antidepressant. Curcumin is the main component of Curcuma longa L, a well-known indigenous herb with comprehensive bioactivities. In the present study, we investigated the effects of chronic unpredictable mild stress (CUMS) and curcumin on behaviours and serotonergic receptor-coupled AC-cAMP signal pathway in rats. Curcumin produced beneficial effects on the stressed rats by effectively improving CUMS-induced low sucrose consumption and reducing serum corticosterone levels in rats. Moreover, curcumin enhanced AC activity and cAMP levels in platelet and various brain regions, and up-regulated mRNA expressions of AC subtypes AC 2, AC 8 and cAMP response element binding protein (CREB) in the hippocampus, cortex and hypothalamus of the CUMS rats. Curcumin also attenuated CUMS-induced reductions of 5-hydroxytryptamine (5 HT) levels and high expressions of central 5-HT(1A/1B/7) receptors in rats. These results suggested that the potent antidepressant property of curcumin might be attributed to its improvement of AC-cAMP pathway as well as CREB via suppressing central 5-HT(1A/1B/7) receptors in the CUMS rats. Our findings provided a basis for examining the interaction of serotonergic receptors and AC-cAMP pathway in depression and curcumin treatment. PMID- 19302829 TI - Effects of the DRD3 Ser9Gly polymorphism on aripiprazole efficacy in schizophrenic patients as modified by clinical factors. AB - Aripiprazole, a novel antipsychotic agent, has a unique pharmacological action (partial agonist) on the dopamine neurotransmission system. Aripiprazole has high affinity for dopamine D2 and D3 receptors (DRD2 and DRD3). We investigated whether the efficacy of aripiprazole can be predicted by a functional DRD3 gene polymorphism Ser9Gly (rs6280) as modified by clinical factors in Han Chinese hospitalized patients with acutely exacerbated schizophrenia. After hospitalization, the patients (n=128) were given aripiprazole for up to four weeks. Patients were genotyped for DRD3 Ser9Gly polymorphism by Restriction Fragment Length Polymorphism (RFLP) method. Clinical factors such as gender, age, duration of illness, education level, diagnostic subtype and medication dosage were recorded. Psychopathology was measured biweekly with the Positive and Negative Syndrome Scale (PANSS). The effects of genetic and clinical factors on PANSS performance after aripiprazole treatment were analyzed by a mixed model regression approach (SAS Proc MIXED). We found that, although the Ser carriers have numerically larger score reductions when compared with non-carriers in almost all PANSS dimensions, the difference of their effects are statically not significant. However, the clinical factors, including dosage of aripiprazole, age, duration of illness, and diagnostic subtype could influence PANSS performance after aripiprazole treatment. This study suggests that DRD3 Ser9Gly polymorphism may not contribute significantly to inter-individual differences in therapeutic efficacy of aripiprazole, but some clinical factors may predict treatment efficacy. PMID- 19302830 TI - Resolution of risperidone-induced tardive dyskinesia with a switch to aripiprazole monotherapy. PMID- 19302831 TI - Prediction of relapse in melancholic depressive patients in a 2-year follow-up study with corticotropin releasing factor test. AB - PURPOSE: To study the power of CRF stimulation test to predict relapse in a sample of melancholic depressive patients in depressed phase, followed-up over a two-year period from the moment they achieved complete remission of depressive symptoms. METHODS: Fifty-one outpatients diagnosed with unipolar depressive disorder with melancholic features according to DSM-IV were assessed with the CRF test. The Structured Clinical Interview for DSM-IV (SCID-IV) was used for diagnosis. Monthly follow-up visits were held over a two-year period after remission; relapse was established using HDRS according to Frank's criteria [Frank E, Prien RF, Jarret RB, Keller MB, Kupfer DJ, Lavori PW, et al. Conceptualization and rationale for consensus definitions of terms in major depressive disorder: remission, recovery, relapse, and recurrence. Arch Gen Psychiatry 1991;48:851-5]. Forty-three patients completed the study. Non controlled antidepressant treatment protocols were used. Predictive statistical analysis was performed through logistic regression. FINDINGS: The final predictive model included three variables: net area under cortisol curve (NAUCC), previous suicide attempt, and stress during follow-up. Sensitivity was of 89%, and specificity was of 92%. NAUCC has shown a predictive power of 80%, with an optimal cut-off point of 251.24 microg/ml/min. CONCLUSIONS: Cortisol is the hormone of the HPA axis which shows the highest power to predict relapse. NAUCC is the most relevant variable. The complete predictive model is a complex combination of biological, clinical and psychoenvironmental variables (NAUCC, previous suicide attempts, and stress during follow-up). Further studies with better control of the psychoenvironmental variables are required to obtain more precise neuroendocrine findings. PMID- 19302832 TI - Activity-dependent plasticity of developing climbing fiber-Purkinje cell synapses. AB - Elimination of redundant synapses and strengthening of the surviving ones are crucial steps in the development of the nervous system. Both processes can be readily followed at the climbing fiber to Purkinje cell synapse in the cerebellum. Shortly after birth, around five equally strong climbing fiber synapses are established. Subsequently, one of these five synaptic connections starts to grow in size and synaptic strength, while the others degenerate and eventually disappear. Both the elimination of the redundant climbing fiber synapses and the strengthening of the surviving one depend on a combination of a genetically coded blueprint and synaptic activity. Recently, it has been shown that synaptic activity affects the synaptic strength of developing climbing fibers. Remarkably, the same pattern of paired activity of the presynaptic climbing fiber and the postsynaptic Purkinje cell resulted in strengthening of already "large" climbing fibers and weakening of already "weak" climbing fibers. In this review, we will integrate the current knowledge of synaptic plasticity of climbing fibers with that of other processes affecting climbing fiber development. PMID- 19302833 TI - Nitric oxide synthase inhibition attenuates L-DOPA-induced dyskinesias in a rodent model of Parkinson's disease. AB - Chronic L-DOPA pharmacotherapy in Parkinson's disease is often accompanied by the development of abnormal and excessive movements known as L-DOPA-induced dyskinesia. Rats with 6-hydroxydopamine lesion of dopaminergic neurons chronically treated with L-DOPA develop a rodent analog of this dyskinesia characterized by severe axial, limb, locomotor and orofacial abnormal involuntary movements. While the mechanisms by which these effects occur are not clear, they may involve the nitric oxide system. In the present study we investigate if nitric oxide synthase inhibitors can prevent dyskinesias induced by repeated administration of L-DOPA in rats with unilateral 6-hydroxydopamine lesion. Chronic L-DOPA (high fixed dose, 100 mg/kg; low escalating dose, 10-30 mg/kg) treatment induced progressive dyskinesia changes. Two nitric oxide synthase inhibitors, 7-nitroindazole (1-30 mg/kg) and NG-nitro-L-arginine (50 mg/kg), given 30 min before L-DOPA, attenuate dyskinesia. 7-Nitroindazolee also improved motor performance of these animals in the rota-rod test. These results suggest the possibility that nitric oxide synthase inhibitors may be useful to treat L DOPA-induced dyskinesia. PMID- 19302834 TI - Effects of benfluorex-vitamin C supplementation on cutaneous capillaries of diabetic rats. AB - The ultrastructural changes on capillaries of the dermis in diabetic and benfluorex-vitamin C treated diabetic rats have been investigated. Three groups of 21 Wistar albino rats were used in the examination: control, diabetes, and benfluorex-vitamin C treated diabetic rats. Diabetes was induced by injection of streptozotocin. The streptozotocin-induced group was treated for 21 days with vitamin C and benfluorex, of which antidiabetic and antihyperlipidemic effects were experimentally proved. Samples taken from the skin of rats' legs were examined under transmission electron microscopy. Swollen endothelial cells, narrowed capillary lumens, a thickened basement membrane, and fusion of mitochondrial cristae in the capillaries of diabetic rat dermis were seen. In the benfluorex-vitamin C treated group, contrary to the diabetic group, neither signs of degeneration in endothelial cells nor a significant difference with the control group with regard to capillary structure were observed. Amelioration in capillaries appears to be due to benfluorex and vitamin C treatment in diabetes. PMID- 19302835 TI - Rapid adaptation of molecular resources from zebrafish and medaka to develop an estuarine/marine model. AB - Many estuary and coastal waters are highly threatened by heavy anthropogenic pollutants. Oryzias melastigma, also called O. dancena, a marine medaka that showed sensitive response to hypoxia and estrogenic endocrine disruptors in previous studies, is becoming a sentinel species for marine ecotoxicology studies. However, the lack of strong molecular foundation and knowledge of early developmental stages hampers its practical applications. Combining our research strength on zebrafish embryos, this study revealed both morphological and molecular (at mRNA and protein levels) development of embryos of this emergent model. Whole mount immunostaining technique specific for O. melastigma was successfully developed based on zebrafish standard protocols. We demonstrated that 17 out of 61 primary antibodies, which were previously tested in zebrafish, showed specific immunoreactivity with O. melastigma. These antibodies clearly illustrated the embryonic development of target tissues (principally neurons) in this medaka. Additionally, partial cDNA fragments of 11 organ-specific marker genes were isolated according to genomic resources of zebrafish, Japanese medaka and other fishes. Of the 11 genes, 8 are widely used as organ markers and their expression patterns were remarkably similar to their homologues in zebrafish and Japanese medaka. The expression profiles of the remaining 3 genes in fish are reported for the first time. These molecular markers (17 antibodies and 11 mRNA probes) can be used as responsive indicators in environmental toxicity evaluation. Moreover, this study brought forward and demonstrated the advantage of transferring techniques and resources from one model to another to hasten the research of interest. PMID- 19302836 TI - Thrombotic cutaneous gangrene with autoamputation of the penis: a rare extracolonic manifestation of ulcerative colitis in a child. AB - We present a pediatric patient with ulcerative colitis who developed thrombotic cutaneous gangrene involving skin of the lower chest, abdomen, back, bilateral buttocks, bilateral thighs, perineum, and genitalia, ultimately resulting in autoamputation of the glans penis. After an extensive review of the literature, we describe the diagnosis and management of this devastating condition. PMID- 19302837 TI - Intraabdominal pulmonary sequestration presenting with elevated urinary normetanephrine levels. AB - Carrying a prenatal diagnosis of a left-sided intraabdominal mass, a term female newborn underwent postnatal imaging that confirmed a left suprarenal mass. Urinary normetanephrine levels were elevated. Given a preoperative diagnosis of neuroblastoma, the baby underwent an uneventful resection of the mass en bloc with the left adrenal gland. The pathologic examination returned pulmonary sequestration and a normal adrenal gland. Postoperative urinary catecholamines were normal. To the best of our knowledge, this is the first description of a newborn with an intraabdominal pulmonary sequestration presenting with elevated urinary catecholamines. PMID- 19302838 TI - Repair of huge incisional hernias intentionally made during infantile living donor liver transplantation. AB - BACKGROUND AND PURPOSE: In some small infants who are transplanted with a large for-size graft by living donor liver transplantation, an incisional hernia is intentionally made to decrease the tension on the graft. The procedure and timing for repair of this type of hernia were retrospectively evaluated. PATIENTS AND METHODS: Repair was carried out in 3 patients at 4 to 11 years after living donor liver transplantation. The preoperative, perioperative, and postoperative statuses were analyzed in each patient. RESULT: Fascial closure was possible in all 3 patients. In 2 patients, separation of a component of the rectus sheath or a lower part of the major pectoral muscle was required for approximation of the fascia. One recipient had transient bile leakage that was treated successfully. CONCLUSION: An intentionally made hernia should be as small as possible to facilitate easy primary closure at a later date. A procedure resembling the "clam shell opening" method, which used a partly separated and extended sheet of the fascia, was feasible to avoid the requirement for an artificial mesh. Preschool ages may be suitable for easier approximation of the fascia. PMID- 19302839 TI - Vasoactive intestinal polypeptide-producing ganglioneuromatosis involving the entire colon and rectum. AB - Vasoactive intestinal polypeptide (VIP) is known to cause the watery diarrhea, hypokalemia, and achlorhydria syndrome. A 14-year-old girl was admitted with a 4 year history of persistent uncontrollable diarrhea and hypokalemia. Computed tomographic evaluation of the neck, chest, and abdomen were normal. Numerous polyps covering the entire colon and rectum were noted on colonoscopy. The serum VIP level was 143 pg/mL. The patient underwent a total proctocolectomy with an ileal-J-pouch. The pathologic examination revealed ganglioneuromatosis. Postoperatively, the symptoms resolved, and the serum VIP level fell to lower than 5 pg/mL. This is an unusual case of the watery diarrhea, hypokalemia, and achlorhydria syndrome caused by ganglioneuromatosis of the entire colon and rectum. PMID- 19302840 TI - Association of gastroschisis and choledochal cyst. AB - Two infants are described who presented in the neonatal period with a direct hyperbilirubinemia. This was initially presumed to be because of the diagnosis of gastroschisis and the prolonged use of parenteral nutrition. However, both infants were eventually found to have an associated choledochal cyst. The cases are a novel association not previously reported and should heighten the awareness that anatomical causes of a direct hyperbilirubinemia need to be ruled out in all neonates. PMID- 19302841 TI - External-internal carotid artery transposition for repair of multiple pseudoaneurysms from penetrating injury in a pediatric patient. AB - A 13-year-old boy was transferred to our trauma center after sustaining a shotgun wound to his neck and head. Workup revealed an injury to his tonsillar fossa, a pseudoaneurysm less than 4 mm in his internal carotid artery, and diffuse cerebral edema. After management of his intracranial hypertension, follow-up angiogram revealed 4 pseudoaneurysms in his internal carotid artery. In the operating room, the affected segment was resected, and a transposition of the external carotid artery to the internal carotid artery was performed. Workup of penetrating neck trauma and management options for internal carotid artery pseudoaneurysms in a pediatric patient are discussed. PMID- 19302842 TI - Manifestation of alveolar rhabdomyosarcoma as primary cutaneous lesions in a neonate with Beckwith-Wiedemann syndrome. AB - We report a rare case of neonatal Beckwith-Wiedemann syndrome (BWS) associated with alveolar rhabdomyosarcoma (RMS). Alveolar RMS was diagnosed on the basis of excisional biopsy. Chemotherapy was initiated and followed by bone marrow transplantation. The patient, who is now 3 years and 11 months of age, is alive 46 months after the initial diagnosis, albeit with disease. We could not detect the PAX3-FKHR or PAX7-FKHR transcripts; however, we could observe hypomethylation of the differentially methylated region of the long QT intronic transcript 1. Thus, neonatal alveolar RMS with BWS may result from an alternate molecular pathway. PMID- 19302843 TI - Treatment with OK-432 for persistent congenital chylothorax in newborn infants resistant to octreotide. AB - Chylothorax is a relatively uncommon condition defined as an abnormal collection of lymphatic fluid within the pleural space. We are reporting the use of OK-432 for treatment of prolonged idiopathic congenital chylothorax in 2 newborn infants who failed to respond to conservative medical therapy, including octreotide injection. PMID- 19302844 TI - Adolescent paraplegia, morbid obesity, and pickwickian syndrome: outcome of gastric bypass surgery. AB - Loss of mobility, such as what occurs as a consequence of spinal cord injury or malformation, is a risk factor for excess weight gain and can confound weight management efforts. Despite well-documented outcomes of bariatric surgery in ambulatory patients, little information is available regarding weight loss surgery in adult or adolescent paraplegic patients. A 15-year-old adolescent boy with a body mass index of 60 kg/m(2) and complete paraplegia caused by spina bifida developed metabolic dysfunction, severe obstructive sleep apnea, and hypoxemia syndrome. In an effort to avoid a tracheostomy for worsening pickwickian syndrome, he was referred for weight loss surgery. Laparoscopic Roux en-Y gastric bypass surgery was safely performed and resulted in loss of 55% of body weight (83.8% excess weight loss) for 2 years. Risk factors for cardiovascular disease markedly improved, and polysomnography demonstrated complete reversal of sleep apnea with substantial subjective improvement in daytime breathlessness and quality of life. Body composition analysis demonstrated preferential reduction in body fat mass compared with lean mass, without detrimental effect on bone mineral density. This case illustrates that paraplegia does not necessarily impair either weight loss efficacy or comorbidity resolution after Roux-en-Y gastric bypass surgery. PMID- 19302845 TI - Pediatric blunt vertebral artery injury: case report and treatment plan. AB - Aggressive screening for blunt cerebrovascular injuries in patients with trauma has led to the identification and successful treatment of these injuries. We report the case of an 8-year-old boy who sustained a vertebral artery injury after a motor vehicle collision. Computed tomography angiogram showed an 8-mm thrombosed segment of the vertebral artery. The patient was initially anticoagulated with a heparin drip and transitioned over to treatment with enoxaparin sodium (Lovenox). With few reports in the literature of blunt cerebrovascular injuries in the pediatric population, a review of the appropriate screening parameters, treatment plans, and follow-up is helpful for the practicing physician. PMID- 19302846 TI - Distance to care and relative supply among pediatric surgical subspecialties. AB - BACKGROUND/PURPOSE: The aim of this study is to describe geographic proximity to and quantify relative supply of 7 pediatric surgical specialties in the United States. METHODS: Data from the 2005 American Medical Association Physician Masterfile and the Claritas Pop-Facts Database were used to calculate subspecialty-specific, population-weighted, straight-line distances between each zip code centroid and the nearest provider. These same data sources were used to calculate the percentage of hospital referral regions with a provider, the percentage of the younger than 18 years population living within selected distances of providers, and provider-to-population ratios for each of the pediatric surgical subspecialties. Further, we calculated the correlation between practice locations and children's hospitals offering pediatric surgical services. RESULTS: Across pediatric surgical specialties, average distances to the nearest provider ranged from 27.1 miles for pediatric surgery to 100.9 miles for pediatric cardiothoracic surgery. The average population-weighted distance to a provider was less than 30 miles for pediatric surgery and pediatric ophthalmology only. For 5 of the 7 pediatric surgical specialties studied, approximately one quarter of the younger than 18 years population lives more than 1-hour drive from a provider. Provider-to-younger than 18 years population ratios range across hospital referral region from 0.04 per 100,000 for pediatric cardiothoracic surgery to 0.97 per 100,000 for pediatric surgery. The correlation between pediatric surgeons and children's hospitals offering services was 0.72. CONCLUSIONS: Although the practice locations of pediatric surgical subspecialties parallel the geographic distribution of children in the United States, large percentages of the younger than 18 years population must travel long distance to receive care from these providers. Large coefficients of variation reveal substantial maldistribution. These findings lay the groundwork for workforce assessments of the pediatric surgical subspecialties and underscore the need for future studies that assess access barriers for children in need of surgical care. PMID- 19302847 TI - Abnormal intrinsic esophageal innervation in congenital diaphragmatic hernia: a likely cause of motor dysfunction. AB - PURPOSE: Patients with congenital diaphragmatic hernia (CDH) often have dilated esophagus and gastroesophageal reflux. Sparse intrinsic esophageal innervation has been described in rats with CDH, but this issue has not been investigated in patients with CDH. The present study tests the hypothesis that innervatory anomalies could account for motor dysfunction in human CDH. METHODS: The esophagi of CDH (n = 6) and control babies dead of other causes (n = 6) were included in paraffin, transversally sectioned, and immunostained with antineurofilament and anti-S-100 antibodies. The proportion of the section surface occupied by neural structures, the ganglionar surface, and the number of neurons per ganglion were measured in 2 to 5 low-power fields from the proximal and distal esophagus with the assistance of image analysis software. Mann-Whitney tests were used for comparing the results using a threshold of significance of P < .05. RESULTS: The percentage of neural/muscle surface was similar in the upper esophagus in both groups, but it was significantly decreased in the lower esophagus of patients with CDH in comparison with controls. There was a relative scarcity of neural tissue in the intermuscular plexus of the lower esophagus. On the other hand, the ganglionar surface and the number of neurons per ganglion were identical in both groups. These results were similar with both immunostainings. CONCLUSION: Intrinsic innervation of the lower esophagus in CDH is abnormal in terms of decreased density of neural structures in the intermuscular plexus. These neural crest-derived anomalies could explain in part the esophageal dysfunction in survivors of CDH. PMID- 19302848 TI - Cholangiocyte secretion of chemokines in experimental biliary atresia. AB - Biliary atresia (BA) is a disease of the newborn that results in obstruction of the biliary tree. The cause of BA remains unknown; however, recent studies using the murine model of biliary atresia have found that rotavirus infection of the biliary epithelial cell (cholangiocyte) triggers an inflammatory response. We hypothesized that rotavirus infection of cholangiocytes results in the release of chemokines, important mediators of the host immune response. METHODS: In vivo, Balb/c pups were injected with rhesus rotavirus (RRV) or saline, and, their extrahepatic bile ducts were microdissected 2, 5, 7, and 14 days after injection. Next, an immortalized cholangiocyte cell line (mCl) was incubated with RRV or serum-free media. Qualitative and quantitative chemokine assessment was performed using enzyme-linked immunosorbent assay, polymerase chain reaction, and immunohistochemistry. RESULTS: In vivo, increased levels of the chemokines macrophage inflammatory protein 2, monocyte chemotactic protein 1, KC and Regulated upon Activation, Normal T Expressed and Secreted were found in RRV infected murine bile ducts. In vitro, infected mCl cells produced increasing amounts of these same chemokines in relation to dose and time. CONCLUSION: These novel results suggest that chemokine expression by RRV-infected cholangiocytes may trigger a host inflammatory process that causes bile duct obstruction. Understanding how viral infection initiates this response may shed light on the pathogenesis of biliary atresia. PMID- 19302849 TI - Laparoscopic excision of choledochal cyst and Roux-en-Y hepaticojejunostomy in symptomatic neonates. AB - PURPOSE: Choledochal cysts require surgical excision, preferably before the onset of cholangitis. Recently, it has become feasible to accomplish the excision laparoscopically in adults and older children. Yet, whether laparoscopic excision of choledochal cyst can be performed safely in symptomatic neonates with choledochal cyst is unclear. We herewith reviewed our experience of laparoscopic excision of choledochal cysts in neonates. METHODS: We managed 9 neonates with choledochal cysts between April 2003 and February 2007. The choledochal cysts were excised laparoscopically. The Roux-en-Y hepaticojejunostomy was fashioned extracorporeally by exteriorizing the jejunum through the extended umbilical port site. End-to-side anastomosis between the common hepatic duct stump and Roux loop was carried out intracorporeally. The patients were followed up for an average of 26 months. RESULTS: The patients presented with jaundice, pale stool, and deranged liver function tests. The diagnosis was confirmed with ultrasonography postnatally. The median operation time was 3.6 hours. There was no operative complication and no conversion. The blood loss was minimal. The recovery was uneventful, and the median hospital stay was 6 days. The liver function tests normalized 3 to 16 weeks postoperatively. No complication was detected at the follow-up visits. CONCLUSIONS: Our preliminary results show that laparoscopic excision of choledochal cyst and Roux-en-Y hepaticojejunostomy in neonates is both feasible and safe. It curtails further complication of the cysts and reverses the derangement of liver function. In addition, the laparoscopic approach minimizes surgical trauma. PMID- 19302850 TI - Congenital stenosis of the hepatic duct at the porta hepatis in children with choledochal cyst. AB - BACKGROUND: The intrahepatic gallstone is known as one of the complications of choledochal cyst. Stenosis of the hepatic duct may contribute to the formation of the stone. This report describes concurrent congenital stenosis of the hepatic duct identified during the primary operation of choledochal cyst. METHODS: Fourteen children were operated on for choledochal cyst. To identify the concurrent stenosis, cholangiography and inspection of the hepatic duct were performed during the operation. In those patients with identified membranous stenosis, the membrane was resected. In those with nonmembranous stenosis, the wide hilar hepaticojejunostomy was performed after hepatic duct plasty. RESULTS: Eleven sites of hepatic duct stenosis were identified, 8 membranous stenoses and 3 relative stenoses without membrane. The membranous stenoses were classified into 4 subtypes as follows: a small central opening, a marginal opening, 2 openings, and a bridgelike structure. At the primary surgery on choledochal cyst, those stenoses were released, and the formation of the gallstone has not been observed in 4 to 11 years in all cases. PMID- 19302851 TI - A new surgical approach to improve gene transfer in liver using lentiviral vectors. AB - PURPOSE: Metabolic inherited liver diseases are attractive targets for gene therapy. Recombinant lentiviruses are very powerful viral vectors able to infect nonmitotic cells. We wanted to develop a new surgical approach to improve gene transfer in adult liver using low viral doses. MATERIALS AND METHODS: Adult rats were injected with 2.108 infectious particles of lentiviral vectors encoding the green fluorescent protein marker gene under control of a liver-specific promoter transthyretin. In the control group (n = 5), gene delivery was performed by inflow intraportal injection. In the surgical group (n = 5), liver was completely excluded from systemic circulation before viral injection in infrahepatic vena cava with high pressure. RESULTS: At day 9, transduction efficiency was 14.35% in the surgical group 3 and 0.39% in the control group (P = .016). At month 2, the number of transduced hepatocytes decreased in the most part of rats, except in half of rats in the surgical group. Antibodies against green fluorescent protein were detected in all rats at month 2, except one in the surgical group. CONCLUSIONS: We developed a new surgical approach allowing an efficient transduction of hepatocytes in adult rats using lentivirus at low viral doses. We have now to control the immune response to permit long-term expression of transgene. PMID- 19302852 TI - Glutamine decreases inflammation in infant rat endotoxemia. AB - Glutamine may have benefits during neonatal sepsis, but its effects on systemic inflammation are unknown. Our aim was to determine whether glutamine affects inflammation in neonatal endotoxemia. Eleven-day rat pups were given intraperitoneal injections of saline (control; C), endotoxin (300 microg/g Escherichia coli lipopolysaccharide) (E), saline with glutamine (2 mmol/g; G), or endotoxin with glutamine (EG). Animals were killed after 2 or 6 hours. Plasma glutamine (mmol/L) was measured enzymatically, and both tumor necrosis factor alpha (pg/mL) and interleukin 10 (IL-10) were measured by enzyme-linked immunosorbent assay. Results, expressed as mean +/- SEM, were analyzed by analysis of variance. Endotoxemia caused a rapid significant decrease in plasma glutamine at 2 hours (C, 0.73 +/- 0.06; E, 0.32 +/- 0.07; mean difference, 0.41 [95% confidence interval {CI, 0.17-0.64}]; P < .001), which was prevented by intraperitoneal glutamine (EG, 0.59 +/- 0.04; mean difference vs E, 0.27 mmol/L [95% CI, 0.03-0.50]; P < .05), indicating glutamine absorption, whereas CG animals had a plasma glutamine of 0.82 +/- 0.07. Tumor necrosis factor alpha was greatly increased by 2-hour endotoxemia (C, 27 +/- 7; E, 2247 +/- 43; mean difference, 2220 pg/mL [95% CI, 2012-2429]; P < .001), and this increase was partly prevented by glutamine (EG, 1991 +/- 91; P < .05 vs E; mean difference, 256; 95% CI, 47-465; P < .05). The effect of glutamine was more pronounced at 6 hours (C, 32 +/- 27; E, 799 +/- 193; EG, 219 +/- 75, C vs E mean difference, 767; 95% CI, 346-1188; P < .001; E vs EG mean difference, 580; 95% CI, 159-1001; P < .01). The IL-10 levels were also greatly increased by 2-hour endotoxemia (C = 55 +/- 21, E = 2429 +/- 58, EG = 1989 +/- 177; C vs E mean difference, 2374; 95% CI, 2740-2008; P < .001; E vs EG mean difference, 440; 95% CI, 74-807; P < .05). Glutamine administration partially prevents the sepsis-induced fall in plasma glutamine levels and reduces the concentration of both proinflammatory and antiinflammatory cytokines. PMID- 19302853 TI - Probiotic acidified formula in an animal model reduces pulmonary and gastric bacterial load. AB - BACKGROUND/PURPOSE: We previously reported that a diet acidified with citric acid effectively reinforces gastric acid protection against bacterial colonization and translocation. In this study, our objective was to examine a biologically acidified formula hypothesized to be more physiologic than formula acidified with free acid. This study was Institutional Animal Care and Use Committee (IACUC) approved and designed to determine whether this diet is better tolerated and equally effective to acidification with citric acid against gut colonization and subsequent bacterial translocation in a premature infant rabbit model. METHODS: A total of 89 rabbit pups born via cesarean delivery 1 day preterm were randomly assigned to 3 feeding groups: Pelargon Nestle at pH 4.55; NAN Nestle, a control diet at pH 7.0 with similar composition; and NAN Nestle acidified in the laboratory with citric acid at pH 4.55. Pups were gavage fed every 12 hours with Enterobacter cloacae challenges of 10 colony-forming units per milliliter of diet per feed and killed on day 3 of life. Lungs, liver, spleen, mesenteric lymph nodes, stomach, and cecum were cultured and quantitatively analyzed for target organism growth and statistically analyzed using chi(2) and Kruskal-Wallis tests. RESULTS: Pelargon, compared to acidified NAN and NAN, significantly reduced the incidence of gastric colonization (15/33 [45%], 21/27 [78%], and 25/29 [86%], respectively; P < .01) and pulmonary colonization (10/33 [30%], 19/27 [70%], 21/29 [72%]; P < .01). Comparing the bacterial logs of colonized groups, the same benefit is observed in the lungs (0.77 +/- 1.22, 1.89 +/- 1.41, 2.12 +/- 1.47; P < .01). Gut colonization and bacterial translocation were equivalent between treatment groups (mesenteric lymph nodes: 10/33 [30%], 11/27 [40%], 8/29 [27%]; spleen: 10/33 [30%], 7/27 [26%], 8/29 [27%]; liver: 10/33 [30%], 6/27 [22%], 9/29 [31%]; cecum: 33/33 [100%], 27/27 [100%], 29/29 [100%]). CONCLUSION: Biologically acidified formula demonstrated superior protection against pulmonary and gastric colonization compared to normal pH and diets acidified with free acid. Its effects may potentially reduce clinical pulmonary infection. PMID- 19302854 TI - Hyperbaric oxygen therapy reduces the severity of necrotizing enterocolitis in a neonatal rat model. AB - INTRODUCTION: Hyperbaric oxygen (HBO) therapy is known to increase oxygen concentration in tissues leading to induction of an adaptive increase in antioxidants, stimulation of angiogenesis, improvement of white blood cell action, and regulation of inflammatory process. Therefore, we tested the potential beneficial effect of HBO in neonatal rat model of necrotizing enterocolitis (NEC). MATERIALS AND METHODS: Thirty newborn Sprague-Dawley rats, provided by the Experimental Research Council, Gulhane Military Medical Academy, Ankara,Turkey, were randomly divided into 3 groups as follows: NEC, NEC + HBO, and control. Necrotizing enterocolitis was induced by enteral formula feeding and exposure to hypoxia after cold stress at 4 degrees C and oxygen. The NEC + HBO group received HBO at 2.8 atmosphere absolute (ATA) for 90 minutes daily for 3 days. The pups were killed on the fourth day, and their intestinal tissues were harvested for biochemical and histopathologic analysis. Blood samples were also obtained from the pups. RESULTS: The mortality rate was highest in the NEC group (3 pups in the NEC group vs 1 pup in the NEC + HBO group). Malondialdehyde and protein carbonyl content were significantly increased, whereas superoxide dismutase and glutathione peroxidase were significantly decreased in the NEC group. All these changes were similar to control levels in the NEC group by HBO treatment. Nitrate plus nitrite (NO(x)) levels and serum tumor necrosis factor alpha were increased in the NEC group and histopathologic injury score and apoptosis index in the NEC group were significantly higher than in the NEC + HBO group. CONCLUSION: Hyperbaric oxygen significantly reduced the severity of NEC in our study. PMID- 19302855 TI - Bombesin can minimize impairments of interstitial cells of Cajal induced by FK506 in small bowel transplantation. AB - PURPOSE: Interstitial cells of Cajal (ICC) are known as intestinal pacemaker cells and express c-kit on their membrane. Previously, we reported that FK506 had neurotoxicity to enteric ganglia, and bombesin (BBS) preserved them against FK506. The aim of this study was to investigate whether ICC was impaired by FK506 and whether ICC was preserved by BBS against FK506. METHODS: Twelve rats underwent allogeneic SBTx heterotopically and were divided into 2 groups as follows: group A underwent SBTx with FK506 and group B with FK506/BBS. All rats were administered FK506 daily. Either BBS or normal saline was infused continuously from day 14 to 28. Analysis of ICC was performed immunohistochemically with c-kit. Interstitial cells of Cajal were evaluated by counting the number of c-kit-positive clusters in each graft. RESULTS: The expression of c-kit accumulated around 60% of PGP9.5-positive enteric ganglia. The number of c-kit-positive clusters in group A was 22.3 +/- 5.5 clusters per cross section (C/CS) and that in group B was 36.3 +/- 5.1 C/CS. Interstitial cells of Cajal were well preserved in group B. There was a significant difference between groups A and B (P <.001). CONCLUSION: Interstitial cells of Cajal were impaired by FK506 in allografts, and BBS could minimize the impairment of ICC against FK506. PMID- 19302856 TI - Massive chylothorax in small babies. AB - PURPOSE: Perceptions on the role and timing of surgical intervention for high output or massive chylothorax in small and premature babies remain varied. We reviewed our experience with this challenging group of patients to help refine our thinking and devise a more consistent strategy for management. METHODS: We conducted a retrospective analysis of all patients in our institutions' neonatal populations diagnosed with chylothorax from June 2000 to April 2008. RESULTS: Of a total of 23 patients (birth weight, 0.54-4.5 kg; gestational age, 23-41 weeks), 17 were treated conservatively, and 6 with massive chylothorax (>50 mL/kg per day) were treated surgically. Surgical treatment varied, including en masse thoracic duct ligation, mechanical pleurodesis, and application of fibrin glue. Survival in the surgically treated group was 83% vs 59% in the conservatively treated group. Median duration of chest tube drainage was 5 days (postoperative, range, 4-16) in the surgically treated group vs 14 days (range, 1-68) in the conservatively treated group. CONCLUSION: Surgery has a definitive role in the care of small babies with massive chylothorax. Daily output exceeding 50 mL/kg per day with no or minimal response to 3 days of maximal medical therapy may indicate a potential therapeutic benefit of surgery. PMID- 19302857 TI - Rising incidence of gastroschisis and exomphalos in New Zealand. AB - BACKGROUND: An apparent increase in the incidence of gastroschisis and exomphalos has been reported from several parts of the world. The exact mechanism of this trend is unknown. The aim of this study was to determine the regional and national trends in the incidence of gastroschisis and exomphalos in New Zealand. MATERIAL AND METHODS: This retrospective multicenter study involved collection of data from all 4 tertiary care pediatric surgical centers in New Zealand. The incidence was calculated per 10,000 live births. Data were analyzed to determine the regional and national trends. The statistical analysis was done using linear regression model and Poisson distribution. RESULTS: The incidence of gastroschisis has increased from 2.96 per 10,000 live births to 5.16 per 10,000 live births between 1996 and 2004. During the same period, the incidence of exomphalos has increased from 0.69 per 10,000 live births to 3.27 per 10,000 live births. Gastroschisis was observed more in younger mothers, whereas exomphalos was associated with older mothers. CONCLUSION: The incidence of gastroschisis and exomphalos is increasing in New Zealand, which is consistent with worldwide trends in showing the increasing incidence of anterior abdominal wall defects. PMID- 19302858 TI - The gonads of 111 South African patients with ovotesticular disorder of sex differentiation. AB - PURPOSE: The aims of this study were to describe the gonadal tissue found in the Southern African true hermaphrodite and establish if there was a correlation between the clinical and histopathologic findings and if these findings were similar to patients with this condition elsewhere. MATERIALS AND METHODS: A retrospective study at the University of KwaZulu-Natal, Durban, South Africa, looked at all patients diagnosed with true hermaphroditism seen between 1984 and 2006. For this 23-year period, 111 consecutive true hermaphrodite patients were diagnosed on clinical findings, internal genital assessments, and the histologic examination of 217 gonadal biopsy specimens. All gonadal tissue taken from these patients was sent for histopathologic evaluation. The results were correlated to the clinical and internal genital evaluations of the patients. RESULTS: Five patients only had a single gonad. Analysis of the gonadal biopsy specimens showed that there were 118 (54%) ovotestes together with 59 ovaries and 40 testes. The ovotestes were divisible on gross appearance into 11% bipolar and 89% mixed types. Histologically, the mixed-type ovotestes have an outer mantle consisting of ovarian tissue, which encapsulated an inner core of 2 distinct types. The first is an admixed ovotestis (constituting 44% of the mixed ovotestes), the central core consisted of gonadal stroma, with scattered foci of separate ovarian and testicular tissue. The second type was the compartmentalized ovotestis (constituting 56% of the mixed ovotestes); here, the outer mantle was thickened in the upper pole and encapsulated a large core of testicular tissue in the lower pole of the gonad. The bipolar ovotestis had a strictly polar distribution of ovarian and testicular tissue, which had an irregularly interdigitating junction between the 2 types of tissue. Statistical analysis showed that no correlation could be found between the type of gonadal tissue and any of the clinical or genital features. CONCLUSION: Three distinct ovotesticular types are identified in the Southern African true hermaphrodite, which have not been described previously. The structure of these gonads has bearing on the type of biopsy done and the subsequent management of the ovotestes. PMID- 19302859 TI - Experimental use of labial mucosa free graft urinary conduit in the Mitrofanoff principle. AB - PURPOSE: One method of treating urinary incontinence is to create a catheterizable and continent vesicostomy by the Mitrofanoff principle, based on the use of a conduit, which in the original method is the appendix. The authors present an experimental technique in which a continent vesicostomy is created by the Mitrofanoff principle, using a conduit made of a labial mucosa free graft. METHODS: Six 30-day-old pigs underwent surgery to create a continent vesicostomy by the Mitrofanoff principle. A tube, made of labial mucosa free graft from the lower lip, was used. The labial mucosal conduit was anastomosed to the bladder mucosa. The tunnel was then prepared, part of which passed through the fibers of the right rectus muscle. The conduit was taken through the tunnel and anastomosed to the skin in the right iliac fossa. RESULTS: Fifty days after the operation, the vesicostomy was continent, pervious, and easily catheterizable in 5 animals. In the sixth pig, the cystostomic tube did not graft successfully because it was infected. CONCLUSIONS: A number of investigators have extrapolated Mitrofanoff method using the ileum, the ureter, or others. The use of a tube made by labial mucosa free graft is an easily performed technique, and as with the Mitrofanoff method, continence and easy catheterization are successfully achieved. PMID- 19302860 TI - The effect of vitamin A deficiency in maternal rats on tumor formation in filial rats. AB - PURPOSE: We established a vitamin A-deficient (VAD) model of pregnant rats to evaluate the effect of vitamin A deficiency in maternal rats on tumor formation in filial rats. METHODS: Ten pregnant Wistar rats were divided into 2 groups: (1) VAD group, 6 rats were given nonvitamin A diet from 2 weeks before mating till delivery and (2) normal diet (ND) group, 4 rats were given normal diet. Twenty random neonatal rats from each group were killed on the next day of delivery. The rest neonates were given normal diet for 1 year until killed. Serum levels of vitamin A, morphology of the kidney, incidence of tumor formation, and retinoid X receptor (RXR) alpha messenger RNA (mRNA) expression in renal tissue were assessed for the filial rats. RESULTS: Fifty-six and 49 neonatal rats were born for VAD group and ND group, respectively. The detection rate of nephrogenic rests (NRs) from neonates in VAD group (50%) was significantly higher than that in ND group (20%; P < .05). The incidence of nephroblastoma was 13.9% in filial rats of VAD group and 0% for ND group. The detection rate of NRs for filial rats of VAD group (30.6%) was significantly higher than that of ND group (6.9%; P < .01). The expression of RXRalpha mRNA in tumor tissue of the filial rats of VAD group (3.17 +/- 0.15) was significantly lower than that in kidney tissue of ND group (3.58 +/ 0.20; P < .01). CONCLUSION: Deficiency in vitamin A for pregnant rats resulted in renal dysplasia, increased NRs, and higher incidence of nephroblastoma. PMID- 19302861 TI - Cranial unifocal Langerhans cell histiocytosis in children. AB - PURPOSE: The present study presents 22 cases of pediatric patients harboring an eosinophilic granuloma (EG) of the skull. METHODS: Twenty-two patients (13 males, 9 females; mean age, 7.5 years; range, 3-14 years) with a suspected diagnosis of a cranial EG were enrolled in the study. They all had been preoperatively evaluated by skull x-ray and computed tomography, whereas 10 patients additionally underwent magnetic resonance imaging. To rule out a multifocal disease, scintigraphy was performed in all cases preoperatively. Surgical excision of the lesions was performed, and EG was proven histopathologically. RESULTS: There was a male predominance. Frontal bone was the most common affected bone. One patient had a multifocal disease. Total excision of the lesion was performed in 19 of 22 patients. No patient received postoperative radiotherapy. In the remaining 3 cases because of the periorbital localization of the EG and the subsequent risk of disfigurement, only a biopsy was performed. These patients were treated with sulfamethoxazole and trimethoprim for 6 months. All 3 lesions were resolved. The follow-up examinations ranged from 6 months to 17 years, with a mean follow-up of 6.2 years, and no tumor recurrence was noted. CONCLUSIONS: We conclude that EG is a benign disease. At the time of diagnosis, a bone scan should be performed to rule out a multifocal disease. Surgical resection is the treatment of choice. Nevertheless, for lesions in which excision can cause cosmetic defects, administration of sulfamethoxazole and trimethoprim after biopsy appeared to be an effective alternative treatment. PMID- 19302862 TI - The positive effect of negative pressure: vacuum-assisted fixation of Integra artificial skin for reconstructive surgery. AB - BACKGROUND: Integra artificial skin (Integra) (Integra Life Sciences Corporation, Plainsboro, NJ) is increasingly used as a skin substitute in reconstructive surgery. However, reliable fixation to the wound bed, a factor of paramount importance for successful application, is often hard to achieve. The vacuum assisted closure system (VAC; KCI, Switzerland), a well-established subatmospheric pressure device, might be of interest to overcome these problems because of its ability to conform to almost any surface. The goal of this study was to test whether negative pressure application yields reliable fixation of Integra in children undergoing reconstructive surgery. METHODS: Between 2001 and 2004, VAC was applied in 18 children (n = 18) aged 7 months to 16.5 years. All required reconstructive surgery with implantation of Integra covering 1% to 12% of the total body surface area. After Integra implantation, VAC was installed for 13 to 30 days. RESULTS: The VAC fixation of Integra was successful in 17 patients (94.5%). The only failure (5.5%) occurred in a patient in whom negative pressure could not be maintained because of a lesion site susceptible to both dislodgement and infection (perianal region). Consequently, infection occurred, and Integra had to be removed. CONCLUSIONS: These results demonstrate that VAC is a valid tool for reliable fixation of Integra in children undergoing even demanding reconstructive surgery. PMID- 19302863 TI - Is the use of cyanoacrylate in intestinal anastomosis a good and reliable alternative? AB - PURPOSE: The present study aims to compare strength, healing, and operation time of experimental intestinal anastomoses performed by polyglactin 910 (Vicryl; Ethicon, Edinburgh, United Kingdom) sutures with ethyl-2-cyanoacrylate glue (Pattex; Henkel, Dusseldorf, Germany). MATERIAL AND METHODS: Ninety-six Sprague Dawley rats were divided into 2 (groups E and L). Each group was further subdivided into 6 subgroups (EA1, EA2, EA3, EB1, EB2, EB3, LA1, LA2, LA3, LB1, LB2, LB3), each containing 8 rats. Intestinal anastomosis was performed by polyglactin 910 sutures in A subgroups and with ethyl-2-cyanoacrylate in B subgroups. The anastomosis was end to end in A1 and B1, side to side in A2 and B2, and end to side in A3 and B3. Time for anastomosis performance (AT) was recorded. In group E, bursting pressures and hydroxyproline levels were determined on the second postoperative day, whereas in group L, the same measurements were made on the sixth postoperative day. One-way analysis of variance was used for analyses of variance in the groups. Quantitative data were analyzed with Student's t test. P value was considered significant at less than .05. RESULTS: There was no significant difference between bursting pressures of subgroup pairs on both postoperative days 2 and 6. Hydroxyproline levels and AT were significantly better in B subgroups. CONCLUSION: Better healing, shorter AT, and equal strength were achieved with ethyl-2-cyanoacrylate compared with polyglactin 910 sutures in intestinal anastomosis in the experimental setting. PMID- 19302864 TI - Outcomes of Hirschsprung's disease associated with Mowat-Wilson syndrome. AB - PURPOSE: Mowat-Wilson syndrome (MWS) is a developmental disorder presenting with mental retardation, delayed motor development, and a wide spectrum of clinical features. Hirschsprung's disease (HD) is associated in almost 50% of cases. This report aims to analyze the course of HD and to evaluate the clinical outcomes of these patients. PATIENTS AND METHODS: Between 1997 and 2007, 110 patients presenting with HD were diagnosed and managed in our institution. Five of them presented the association of HD and MWS. Their records were reviewed retrospectively. RESULTS: All of the 5 patients have a genetic disorder specific of MWS (nonsense mutation or deletion on SIP1 gene, locus 2q22). Two patients underwent transanal endorectal pull-through procedure for classic rectosigmoid HD. Three patients were operated on for total colonic aganglionosis using Duhamel procedure. The median follow-up was 4 (range, 0.3-7) years. Only one patient is doing well (rectosigmoid HD). Two patients have a stoma diversion for severe motility disorders. Of the 3 total colonic aganglionosis, one still has repeated episodes of obstruction requiring total parenteral nutrition (TPN). The 2 others still have repeated episodes of enterocolitis. All patients required a prolonged TPN (32.5 months in average). CONCLUSION: Hirschsprung's disease associated with MWS is a severe condition. Even in case of short segment HD, patients can present motility disorder requiring a prolonged TPN. Physician and surgeon should be aware about the evolution of this rare condition. PMID- 19302865 TI - Expression of EphB2 in the development of anorectal malformations in fetal rats. AB - PURPOSE: The receptor tyrosine kinase of the Eph family is a large group of highly conserved molecules that function in diverse intercellular recognition events. It has been reported that EphB2 is related to caudal remodeling events. The aim of this study is to investigate EphB2 expression in anorectal development in normal and rat embryos with anorectal malformations (ARMs) and attempt to define its role in anorectal morphogenesis. METHODS: The ethylenethiourea (ETU) rat model of the ARMs was used in this study. Immunohistochemical analyses and real time quantitative polymerase chain reaction (PCR) were carried out to investigate EphB2 protein localizations and messenger RNA (mRNA) expression levels. (1) Rat embryos with ARMs were obtained by treating pregnant rats (n = 24) with administration of ETU on gestation day (Gd) 10. Normal rat embryos (n = 111) and embryos treated by ETU without ARMs (n = 90) were the control groups, and embryos with ARMs (n = 108) from Gd13 to Gd16 were divided according to the sections taken from specimens. (2) Embryos were sequentially sectioned in the sagittal and transversal planes before staining with a specific antibody to EphB2. Spatiotemporal study was carried out on EphB2 expression. (3) Individual frozen sections were used to manually microdissect the cloaca and anorectal specimens for total RNA extraction. EphB2 expression was evaluated by real time quantitative PCR. RESULTS: On the immunologic labeling study, EphB2 expression was confined to the cloaca in control groups, whereas EphB2 expression was mainly located at the urorectal septum (URS) and cloacal membrane on Gd13 and Gd14. The increased positive expression was observed in the fused tissue of the URS and cloacal membrane on Gd15. On Gd16, the anal membrane broke down, and the rectum was able to be in contact with the anus, and EphB2 expression was then noted in mucous membrane of rectum. EphB2 expression was seen in the cloacal and anorectal tissues of embryos with ARMs. By integrated optical density (IOD) measurement, IOD value of EphB2 protein was significantly lower in the ARM group than that in the control groups on Gd13 to Gd16 (P < .05), respectively. As shown by real time quantitative PCR, EphB2 expression was detected in 3 groups. EphB2 mRNA level increased on Gd13 to Gd16 but gradually decreased after Gd16. The expression level of EphB2 mRNA in the ARM embryos was lower on Gd13 to Gd16 than that in control groups (P < .05). CONCLUSIONS: EphB2 expression decreased in the ARM embryos and was confined to URS and cloaca, whereas it was higher in control group. Our data thus indicated that EphB2 molecules possibly contributed to the anorectal morphogenesis and the decreased expression of EphB2 might be related to the development of ARMs. PMID- 19302866 TI - Congenital pouch colon associated with anorectal malformation-histopathologic evaluation. AB - BACKGROUND: Although enough literature is available on the descriptive and the management aspects of congenital pouch colon (CPC) associated with anorectal malformation (ARM), there is paucity of its detailed histopathologic studies. The present article details the histopathologic observations in the excised specimens of CPC in the neonates operated on for ARM. MATERIAL AND METHODS: Formalin-fixed specimens of CPC were evaluated both grossly and microscopically. They were stained by H&E stain and Masson trichrome stain. RESULTS: Haustrations and tenia coli were uniformly absent in all the CPC specimens. Mucosa showed focal erosion, hemorrhage, and disruption in the muscularis mucosae. Submucosa showed congested blood vessels, hemorrhage, and thrombosis. Both the circular and the longitudinal muscle layers showed hypertrophic and atrophic appearance. Both the muscle layers also showed disruption. There was presence of fibrosis in all the muscular layers on Masson trichrome stain. There was presence of unique constriction bands in the muscularis propria. These were seen as focal dipping and narrowing of the muscular layers at intermittent intervals. They were more prominent in the circular muscle layers. CONCLUSION: Congenital pouch colon seen in association with ARM is an abnormally developed tissue and needs to be resected for better functional outcome of the remaining gut. However, further physiologic and immunohistochemical studies are warranted to observe the contractile and innervation pattern of CPC. PMID- 19302867 TI - The lung primordium an outpouching from the foregut! Evidence-based dogma or myth? AB - BACKGROUND/PURPOSE: This study examines and challenges the "evidence-based legitimacy" of the theory, "the lung primordium is an outpouching from the foregut." METHOD: A literature review was undertaken using computer database, journals, and relevant anatomical and embryological texts. RESULTS: The independent path of development taken by the tracheobronchial system and the oesophagus once identified as separate entities; the lack of morphologic, molecular, biological, and genetic supportive evidence for the "common-origin" theory; the distinct longitudinal line of demarcation between the nonsegmented muscles of the esophagus and the highly segmented cartilaginous structure of the tracheobronchial tree; the absence of a tracheoesophageal septum in the process of separation; the differences in epithelial lining; and the diametrically opposed mucociliary cascade of the upper airway vs the mucociliary escalator of the tracheobronchial tree all seriously challenge the authenticity of a common origin to these 2 entities. CONCLUSION: To the extent that the foregut is seen as consisting of 2 separate semitubular splanchnopleuric entities ventrodorsally juxtaposed, it is true that the lung primordium as an outpouching of, and not from, the foregut. This must never be confused with the notion that the esophagus and tracheobronchial tree have a common origin. In fact, they develop from 2 completely separate segments of the trilaminar germ disk, but because of head fold development are brought together to create a common tracheoesophageal chamber that is later separated, facilitated by the prochordal membrane diverticulum. PMID- 19302868 TI - Chronic inflammation in congenital cystic adenomatoid malformations. An underestimated risk factor? AB - PURPOSE: Congenital cystic malformations of the lung are more frequently diagnosed before birth, but guidelines for surgical management of asymptomatic cases are lacking. The aim of this article is to review our 10-year results with antenatally diagnosed congenital cystic adenomatoid malformations (CCAMs) to debate indications for early postnatal surgical management in asymptomatic patients. METHOD: Twenty-four cases were reviewed; of these, 18 were operated on before 15 days of life for respiratory distress or mediastinal shift, whereas 6 were submitted to elective surgery at 3 months of age. RESULTS: Twenty lobectomies and 4 atypical resections were performed. Two of the latter required a second surgery for incomplete primary perinatal resection. No postsurgical complications were reported. Nineteen (19/24) of the resected specimens showed signs of chronic inflammation. In the perinatal period, 100% (8 cases) of CCAM type II and 50% (8 cases) of CCAM type I resulted to be inflamed. Of the asymptomatic cases, 50% (3/6) were also found to be affected. No infections were detected at bacteriologic culture and bacterial debris was stained in 3 specimens. CONCLUSION: In this series, a 79% incidence of pulmonary inflammation was detected. The CCAM type II resulted to be always involved in this process of inflammation. This was an unexpected finding, particularly in cases without mediastinal shift or respiratory distress. In light of these results, early postnatal treatment, at around 3 to 6 months of age, could be considered even in asymptomatic patients. PMID- 19302869 TI - Myectomy-myotomy for long segment Hirschsprung's disease in a patient with Haddad syndrome. AB - Hirschsprung's disease is a rare entity with an incidence of 1 in 5000 live born infants. Long segment Hirschsprung's disease occurs in approximately 5% to 10% of this patient population and is defined as a transition zone proximal to the sigmoid colon (Bodian M, Carter CO, Ward BC. Hirschsprung's disease. Lancet. 1951;1:302-309). The association of congenital central hypoventilation syndrome (also known as Ondine's curse) and Hirschsprung's disease is termed Haddad syndrome. PMID- 19302870 TI - From a branchial fistula to a branchiootorenal syndrome: a case report and review of the literature. AB - Branchial abnormalities constitute 32% to 45% of all neck pathologies in children. They may be a part of branchiootorenal (BOR) syndrome, which is characterized by branchial arch anomalies, preauricular pits, hearing impairment, and renal malformations. Typically, the management of a branchial fistula does not necessarily require an extensive diagnostic workup. However, in patients with a branchial fistula associated with external ear anomalies on physical examination and/or a history of hearing loss and a similar history and findings in other family members, an additional workup should be performed to eliminate the possibility of BOR syndrome. The aim of this report is to make pediatric surgeons aware of the BOR syndrome in patients presenting with branchial arch anomalies. PMID- 19302871 TI - An unusual presentation of a fourth pharyngeal arch (branchial cleft) sinus. AB - Fourth pharyngeal arch (branchial cleft) sinuses and fistulas are a rare cause of recurrent neck abscesses presenting in the first 2 decades of life. Because they are uncommon, the diagnosis and subsequent treatment can be delayed, even with proper radiographic imaging and clinical evaluation. This case illustrates a life threatening presentation of a ruptured fourth branchial cleft sinus with extravasation into the mediastinum, chest, and abdomen. PMID- 19302872 TI - Congenital biliary atresia: liver injury begins at birth. AB - BACKGROUND: The timing of onset of liver injury in biliary atresia (BA) is not known, although in approximately 10% of cases, biliary pathologic condition associated with the biliary atresia splenic malformation syndrome must begin well before birth. METHODS: The study involved retrospective case-note review for infants with definite BA who underwent laparotomy within first week of life. RESULTS: Three infants were identified who had occlusive BA evident on the first day of life. In all cases, their liver was grossly normal, and histologic changes were trivial. CONCLUSION: This suggests that the detrimental cholestatic liver injury, later characteristic of BA, only begins from the time of birth despite a prenatal occlusive biliary pathology. It may be that tissue injury only occurs with the onset of the perinatal bile surge initiating periductal bile leakage and the triggering of an inflammatory and ultimately fibrotic response. PMID- 19302873 TI - Duodenal perforation because of swallowed ballpoint pen and its laparoscopic management: report of a case. AB - Accidental ingestion of foreign objects by teenagers or young adults without mental impairment is a rare event. We present a case of a patient who unintentionally swallowed a ballpoint pen several days before seeking medical attention. She concealed the event until abdominal radiographs revealed the foreign object. Because swallowed foreign objects can be potentially harmful, they should be removed endoscopically as soon as possible to prevent development of complications. At times, they need to be removed operatively. This report describes the laparoscopic removal of a ballpoint pen that perforated the duodenum. PMID- 19302874 TI - Fibromatosis presenting as a sacrococcygeal mass. AB - Most children older than a year who present with a sacrococcygeal mass have a malignant tumor. We describe an 8-year-old child with of a sacrococcygeal soft tissue mass that was completely resected with histologic findings consistent with fibromatosis. Fibromatosis is a deep-seated, musculoaponeurotic, borderline tumor that is rare in childhood and has a high incidence of local recurrence even after margin-free resection. We review and discuss management strategies for treatment of fibromatosis in children. This case represents the first report of fibromatosis presenting as a sacrococcygeal mass. PMID- 19302875 TI - Long segment congenital tracheal stenosis in twins successfully treated by slide tracheoplasty. AB - Long segment congenital tracheal stenosis (LSCTS), associated with complete tracheal rings, is a rare condition, difficult to manage and historically associated with high mortality rate. We report two pairs of identical twins all affected by LSCTS successfully treated by sliding tracheoplasty. All had severe respiratory distress. Three infants had left pulmonary artery (LPA) sling and one intra-cardiac malformation. Slide tracheoplasty was done under cardiopulmonary bypass, and cardiovascular malformations were corrected at the same time. One child needed plication of paralyzed right hemi-diaphragm and another distal tracheal Palmaz stent insertion due severe tracheobronchomalacia. All children are doing well during 6 months follow-up. Slide tracheoplasty seems to produce the same good early results in twins as for singletons with LSCTS. PMID- 19302876 TI - Severe injuries from coin cell battery ingestions: 2 case reports. AB - Although coin cell battery ingestion is usually not associated with adverse consequences, case reports have been published that describe serious morbidity and occasional mortality. This report describes 2 young children developing serious complications from unwitnessed Lithium coin cell ingestion. A 19-month old developed an aortoesophageal fistula from the proximal descending aorta, whereas the other developed bilateral vocal cord paralysis. Massive bleeding from the aortoesophageal fistula occurred 10 days post battery removal, resulting in a fatal outcome despite maximal surgical efforts. The patient with bilateral vocal cord paralysis required tracheostomy for airway stridor but is recovering function. This report adds to the evidence that primary prevention efforts are needed to caution parents about the dangers associated with coin cell batteries, and secondary prevention can be improved by raising awareness among clinicians as to the various clinical presentations and therapeutic options for this condition. PMID- 19302877 TI - Tracheobronchial injury diagnosed by computed tomography scan: a case report. AB - Tracheobronchial injuries are uncommon after pediatric blunt trauma. Because they are easily missed and potentially life-threatening, surgeons caring for pediatric patients with trauma should be aware of this spectrum of injuries. We present a case of tracheobronchial injury in a 10-year-old girl caused by blunt trauma sustained during an all-terrain vehicle collision. This injury was diagnosed in a timely fashion with computed tomography scan, and operative repair was accomplished. A discussion of the management of tracheobronchial injury with brief literature review follows. PMID- 19302878 TI - Gastroschisis: effect of mode of delivery. PMID- 19302881 TI - Stem cells as a potential treatment of necrotizing enterocolitis. PMID- 19302882 TI - Epidemiological puzzle pieces. PMID- 19302883 TI - The relationship between abdominal fat, obesity, and common mental disorders: results from the HUNT study. AB - OBJECTIVE: While recent studies have suggested an association between obesity and common mental disorders, findings across different studies have varied and the nature of any relationship remains uncertain. The aim of this study was to examine the relationship between depression, anxiety, and two different measures of obesity in a large community-based sample. METHODS: The HUNT-2 study was used to conduct a cross-sectional study of 65,648 adults between 20 and 89 years of age. During a clinical examination, trained nurses took various anthropometric measurements allowing participants' body mass index (BMI) and waist-hip ratio (WHR) to be calculated. Anxiety and depressive symptoms were ascertained using the Hospital Anxiety and Depression Scale. Information regarding physical activity, level of social support, and medical comorbidity was also obtained. RESULTS: Elevated WHR was associated with increased prevalence of both anxiety and depression. After adjustment for BMI, physical activity, social isolation, and somatic diseases, WHR remained independently associated with depression in both males and females and with anxiety among males. Obesity, as defined by BMI, was associated with depression; however, this association was strongly attenuated by WHR. There was a negative association between BMI and anxiety in both genders. CONCLUSION: Abdominal fat distribution (as measured by WHR) appears to be the key mediator in the relationship between obesity and depression. Increased BMI was not independently associated with depression and may provide some protection against anxiety. These findings are consistent with a hypothesis that links obesity and depression via metabolic disturbances involving the hypothalamic pituitary-adrenocortical axis. PMID- 19302884 TI - Associations of obesity with psychiatric disorders and suicidal behaviors in a nationally representative sample. AB - OBJECTIVE: To determine whether obesity is associated with a variety of psychiatric outcomes after taking into account physical health conditions. METHODS: Data came from the public use dataset of the Canadian Community Health Survey Cycle 1.2 (age 15 years and older, N=36,984). Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition psychiatric diagnoses of major depressive disorder, mania, panic attacks, panic disorder, social phobia, agoraphobia, alcohol dependence, and drug dependence were examined, as was suicidal behavior (ideation or attempts). Multiple logistic regression was utilized to examine the association between obesity (defined as body mass index >or=30) and mental health outcomes. Covariates in the regressions included sociodemographic factors and a measure of physical illness burden (the Charlson Comorbidity Index). RESULTS: In adjusted models, obesity was positively related to several lifetime psychiatric disorders (depression, mania, panic attacks, social phobia, agoraphobia without panic disorder), any lifetime mood or anxiety disorder, suicidal ideation, and suicide attempts [adjusted odds ratio (AOR) range: 1.22-1.58]. Obesity was similarly positively associated with past-year depression, mania, panic attacks, social phobia, any anxiety disorder, and suicidal ideation (AOR range: 1.24-1.52), and negatively associated with past year drug dependence (AOR=0.53, 95% CI 0.31-0.89). Most of these associations were found to be specific to women, while some were also present in men. CONCLUSION: Independent of physical health conditions, obesity was associated with psychiatric disorders and suicidal behavior in the Canadian population. Possible mechanisms and clinical implications of these findings are considered. PMID- 19302885 TI - A consultation with genetic information about obesity decreases self-blame about eating and leads to realistic weight loss goals in obese individuals. AB - OBJECTIVE: This study tested the effects of a consultation using genetic information about obesity on attitudes relating to weight loss goals, self-blame about eating, and weight-related coping in obese individuals. Furthermore, the study sought to explore possible predictors for weight gain/loss. METHOD: A total of 411 obese individuals were randomly assigned to two standardized consultations, with and without genetic information about obesity, and a control group without any intervention. After a 6-month follow-up, 253 obese individuals of the intervention groups and 98 individuals of the control group had a complete dataset. Data were analyzed regarding the independent variables assessment time, treatment group, and the familial predisposition (at least one obese parent or sibling). As dependent measures, attitudes about weight loss goals, weight related self-blame, coping, and body shame were assessed via questionnaire or interview. RESULTS: Individuals with and without a familial predisposition profited in different ways from a consultation using genetic information about obesity: at follow-up, individuals with a familial predisposition reported mainly a relieving effect (less self-blame about eating). Both groups reported an adjustment to more realistic weight loss goals and a greater satisfaction with a 5% weight loss. Furthermore, the more negative obese individuals felt about their current weight at baseline, the higher the risk that these individuals had gained weight at follow-up. CONCLUSION: A consultation focusing on genetic factors might be helpful for obese individuals regardless of their familial predisposition, but only predisposed individuals showed a decrease in self-blame about eating. Negative thoughts and feelings about current weight might predict future weight gain. PMID- 19302886 TI - Body image dissatisfaction as an important contributor to suicidal ideation in Korean adolescents: gender difference and mediation of parent and peer relationships. AB - OBJECTIVES: This study was to examine not only the contribution of body image dissatisfaction to suicidal ideation, according to gender, but the mediating role of parent and peer relationships on the association between body image dissatisfaction and suicidal ideation. METHODS: Data from the 2004 Korean Education and Employment Panel, a school-based survey administered to a nationally representative sample of Korean adolescents, are used in the study. With the exception of participants with missing data on important questions (suicidal ideation, self-evaluated body image, self-reported weight/height, risk factors related to suicide), information from 5426 students (2896 males and 2530 females) in grades 9 and 12 was separately analyzed along gender lines using multivariate logistic regression. RESULTS: In both genders, body image dissatisfaction contributed to suicidal ideation, after controlling for covariates (i.e., school type, perceived family economic hardship, parent-related negative life events, delinquent behaviors, unhealthy behaviors, and self efficacy). Such association existed throughout all body mass index ranges, and underweight males and normal females were most vulnerable to suicidal ideation if they are dissatisfied with their bodies. Parent and peer relationships partially mediated the association between body image dissatisfaction and suicidal ideation in both genders, but these effects were very small. CONCLUSIONS: How adolescents evaluate their body image may be an important indicator of suicidal ideation. Future research should further test the link between body dissatisfaction and suicidal ideation, even after controlling for other important forms of psychopathology known to be associated with these issues. PMID- 19302887 TI - Adolescent depressive symptoms and smoking behavior: the gender-specific role of weight concern and dieting. AB - OBJECTIVE: Increased weight concern and dieting are associated with both depression and smoking among adolescents, particularly girls. This cross sectional study examined the gender-specific role of weight concerns and dieting on the adolescent depression-smoking association. METHODS: Participants were 2051 adolescents (1056 boys and 995 girls) from seven high schools in the Netherlands who filled out questionnaires on smoking, depression, and weight constructs and of whom weight and height were measured. RESULTS: Logistic regression analyses showed that for girls, but not for boys, the depression-smoking association disappeared after controlling for weight concerns and dieting. CONCLUSION: Our findings indicate that weight concerns and dieting are important factors to consider in the depression-smoking initiation relation for girls, but not for boys. This suggests that smoking prevention programs for depressed girls might be enhanced by challenging the idea of smoking as a diet strategy and incorporating a focus on healthy weight regulation. However, before setting up a prevention trial, future prospective studies should first examine why and how weight concerns and dieting are important factors in the depression-smoking initiation relation among girls, also in conjunction with other established risk factors such as parental and peer smoking. PMID- 19302888 TI - The reciprocal relationship between parent-child connectedness and adolescent emotional functioning over 5 years. AB - PURPOSE: The purpose of this study was to evaluate the reciprocal relationship between parent-child connectedness and depressive symptoms, self-esteem, and body satisfaction over 5 years in a diverse sample of 2516 male and female adolescents. METHODS: Youth completed Project Eating Among Teens surveys at Time 1 (1998-1999) and Time 2 (2003-2004). Multiple linear regression analysis was conducted to evaluate relationships between parent-child connectedness and adolescents' emotional functioning, controlling for baseline parent-child connectedness. The reciprocal relationship was also evaluated using the same methods. RESULTS: Parent-child connectedness was associated with increased body satisfaction for females, increased self-esteem for males, and decreased depressive symptoms for both males and females. The reciprocal relationship results showed that, among females, self-esteem was associated with increased parent-child connectedness while depressive symptoms predicted decreased parent child connectedness. In males, body satisfaction was associated with increased parent-child connectedness. CONCLUSIONS: Parent-child connectedness and youth emotional functioning reciprocally influenced each other over the 5-year period of this study. Interventions aimed at strengthening the parent-child relationship throughout adolescence may protect emotional health and prevent longer-term emotional consequences in young adults. PMID- 19302889 TI - Alcohol problems and all-cause mortality in men and women: predictive capacity of a clinical screening tool in a 21-year follow-up of a large, UK-wide, general population-based survey. AB - OBJECTIVE: While the relation between alcohol consumption and mortality has been well explored, little is known about the link between alcohol problems and mortality in general population-based studies, particularly among women. This was the objective of the present study METHODS: In this prospective cohort study, 5333 non-abstaining individuals (2539 women) from the UK-wide Health and Lifestyle Survey (aged 42.9 years at study induction) completed the CAGE questionnaire of alcohol problems and participated in a medical examination in 1984/1985; they were then followed up for mortality experience until 2005. RESULTS: Alcohol problems at baseline were less common in women (2.4%) than in men (7.8%). A total of 21 years of follow-up gave rise to 1201 deaths. Elevated rates of mortality were evident in persons reporting symptoms of alcohol problems in comparison to those who did not. In gender-stratified analyses, alcohol problems were more strongly associated with mortality risk in women (age-adjusted hazards ratio: 2.25; 95% confidence interval: 1.22-4.12) than in men (1.49; 1.12 1.99), although this effect modification was not statistically significant (P value for interaction=0.125). Controlling for a range of covariates--including socioeconomic position, co-morbidity (somatic and psychiatric), and alcohol intake--had essentially no impact on these associations. CONCLUSION: The CAGE questionnaire may have some utility in routine health assessments in the general population. PMID- 19302890 TI - Gender and the nocebo response following conditioning and expectancy. AB - OBJECTIVE: To investigate the role of Pavlovian conditioning and expectancy and of gender on the nocebo effects. METHODS: Conditioning experiment: Forty-eight healthy male and female volunteers were investigated for 3 days using a standard rotation procedure. Subjects in the experimental group received a salient oral stimulus prior to rotation; subjects in the control group received the stimulus 12 h after rotations on Days 1 and 2; on Day 3, all subjects received the stimulus prior to rotation. Expectancy experiment: Another 48 healthy subjects were rotated 5 x 1 min once only. All subjects received the same oral stimulus immediately prior to rotation; subjects in the experimental group were told that the symptoms might worsen with the stimulus; controls did not receive additional information. In both experiments, symptom rating (SR) and rotation tolerance (RT) were determined. RESULTS: Conditioning significantly reduced RT (P=.015) and increased SR (P=.024). For both RT and SR, a significant "day x group x gender" effect was found (P=.044; SR: P=.011) indicating that conditioning was more effective in women. Expectancies lowered RT (P=.085) without affecting SR. There was a significant "rotation x gender" interaction on RT (P=.005) indicating that the expectancy was more effective in men. CONCLUSION: Women responded stronger to conditioning while men responded to expectancies, but to a lesser degree. It needs to be determined whether this is restricted to nausea-specific conditions or can be generalized across clinical and experimental conditions. PMID- 19302891 TI - Three job stress models/concepts and oxidative DNA damage in a sample of workers in Japan. AB - OBJECTIVE: Three job stress models/concepts (the job demands-control [DC] model, the effort-reward imbalance [ERI] model, and organizational justice) have been linked to coronary heart disease (CHD) at work. In recent years, oxidative DNA damage has been identified as a new risk factor for CHD. However, evidence for the association between these job stressors and oxidative DNA damage is limited. The present cross-sectional study investigated the association between these job stress models/concepts and oxidative DNA damage as a possible mediator of the adverse health effects of job stress. METHODS: A total of 166 male and 51 female workers of a manufacturing factory in Japan were surveyed using a mailed questionnaire regarding job stressors and demographic, occupational, and lifestyle variables. Urinary concentrations of 8-hydroxy-2'-deoxyguanosine (8 OHdG), a biomarker of oxidative DNA damage, were also measured. RESULTS: In male subjects, the urinary concentrations of 8-OHdG were significantly higher among the group with lower interactional justice, one of the two components of organizational justice; however, no association was observed with the DC model or the ERI model. In female subjects, high job demands/control ratio was significantly and positively associated with the urinary concentrations of 8 OHdG. CONCLUSION: Interactional justice among male workers and the DC model-based strain among female workers may be associated with increased urinary concentrations of 8-OHdG which possibly reflects oxidative DNA damage. PMID- 19302892 TI - Recognition and treatment of depression in primary care: effect of patients' presentation and frequency of consultation. AB - OBJECTIVE: Primary care physicians (PCPs) are expected to recognize depression and appropriately prescribe antidepressants. This article investigated the single and combined effects of different patient presentations and frequency of visits on detection and antidepressant use. METHODS: Data came from an Italian nationwide survey on depressive disorders in primary care, involving 191 PCPs and 1910 attenders. Two hundred fifty patients suffering from major or subthreshold depression were compared in relation to their presentation (psychological, physical, and pain) and frequency of visits (low and high). RESULTS: Recognition of depression significantly varied according to both presentation and frequency of visits. When compared to patients with psychological complaints, the odds ratios for nonrecognition of depression were higher for patients presenting with physical symptoms [2.3; 95% confidence interval (CI)=1.1-5.3] and with pain (4.1; 95% CI=1.6-9.9). Subjects who rarely attended the practice were 2.3 times less likely to receive a diagnosis of depression, compared with those having a high frequency of visits (95% CI=1.2-4.6). Similarly, patients presenting with physical symptoms or with pain and those with a low frequency of visits were rarely treated with antidepressants. The combination of physical or pain presentation with low frequency of visits further increased the risk for nonrecognition, which was sixfold that of the reference category. CONCLUSIONS: Some subgroups of depressed patients still run a high risk of having their depression unrecognized by the PCP. Screening for depression among patients presenting with pain might be useful in order to improve recognition and management. PMID- 19302893 TI - Association of psychosocial factors and bullying at individual and department levels among naval military personnel. AB - OBJECTIVE: We studied the associations between bullying at work and psychosocial factors both at the individual and at the department levels. METHODS: The data were collected in a cross-sectional study from a questionnaire sent by mail in 2002 to all military personnel in the Royal Norwegian Navy as part of a general work and health study. Self-experienced and observed bullying as well as scores for psychosocial scales using the General Nordic Questionnaire for Psychological and Social Factors at Work were calculated both for the individuals (n=1604) and as a mean for each Navy department (n=97). RESULTS: Low scores on the fair leadership, innovative climate, and inequality scales were associated with high occurrence of bullying at the individual level in a backward stepwise multivariate logistic regression, which means that lack of fair leadership and innovative climate and unequal treatment were associated with high occurrence of bullying. At the department level, the three scales were similarly significantly associated with percentage observed bullying when analyzed separately. A low score on the fair leadership scale and lower departmental mean age were associated with high percentage observed bullying in a stepwise multiple linear regression analysis. Repeating the analyses excluding those being bullied did not change the estimates markedly. CONCLUSION: Our findings indicate that the work environment as perceived by the individual and at the department level is related to the occurrence of bullying. PMID- 19302894 TI - Adaptation of clinical practice guidelines on major depressive disorders in the general hospital: an update. PMID- 19302895 TI - [A 77 years-old woman with a leg paraesthesia for one year]. PMID- 19302896 TI - Gabapentin for the treatment of hot flashes in women with natural or tamoxifen induced menopause: a systematic review and meta-analysis. AB - BACKGROUND: Various nonhormonal agents have been used for the treatment of hot flashes in women with natural or tamoxifen-induced menopause. Some studies have reported that gabapentin appears to be an effective and well-tolerated treatment modality. OBJECTIVE: To investigate the efficacy and tolerability of gabapentin for the treatment of menopausal hot flashes, we performed a systematic review of all trials reporting on the efficacy and tolerability of gabapentin in women with hot flashes and a meta-analysis of the randomized controlled trials (RCTs) conducted in this patient population. METHODS: For the systematic review, a literature search was conducted through MEDLINE, EMBASE, and Cochrane Central Register of Controlled Trials for articles published in English from inception of the databases through November 2008. The reference sections of retrieved articles were searched, and a manual search of key journals and abstracts from major meetings in clinical pharmacology was conducted. To be included in the meta analysis, RCTs had to compare gabapentin with placebo in the treatment of hot flashes in women with natural or tamoxifen-induced menopause, regardless of the sample size, dosage used, duration of treatment, or frequency of the episodes. Uncontrolled and openlabel trials were reviewed but excluded from the meta analysis. The percent reduction in hot flash frequency (relative to baseline) and the composite score (summation of the number of hot flashes in each severity category multiplied by the severity score) were used as primary outcome measures. Dropout rates and the incidences of frequently reported adverse events (eg, dizziness/unsteadiness, fatigue/somnolence) were also investigated. RESULTS: The systematic review included 7 trials conducted in 901 patients between 2002 and 2008. Study sizes ranged from 22 to 420 patients, total daily doses of gabapentin ranged from 900 to 2400 mg, and titration periods lasted 3 to 12 days. All of the trials were conducted in North America (6 in the United States and 1 in Canada); 4 of the trials enrolled subjects with a history of breast cancer, whereas the remaining 3 trials only enrolled postmenopausal women. Four RCTs were included in the meta-analysis. Data were expressed as weighted mean difference (WMD) or relative risk (RR), with the associated 95% CI. Women assigned to gabapentin reported a significantly greater percent reduction in both the frequency of hot flashes (WMD = 23.72 [95% CI, 16.46-30.97]; P < 0.001) and the composite score (WMD = 27.26 [95% CI, 21.24-33.29]; P < 0.001), with significant between-study heterogeneity (I(2) = 97.8% and 95.6%, respectively). Dropouts due to adverse events were more frequent in women randomized to gabapentin than in controls (RR = 2.09 [95% CI, 1.13-3.85]; P = 0.02; I(2) = 0%). The risk of symptom clustering also was significantly higher in the treatment group than in the controls (dizziness/unsteadiness: RR = 6.94 [95% CI, 3.19-15.13]; P < 0.001; I(2) = 63.1%; and fatigue/somnolence: RR = 4.78 [95% CI, 2.23-10.25]; P < 0.001; I(2) = 0%). CONCLUSIONS: Comparisons of gabapentin and placebo revealed reductions of 20% to 30% in the frequency and severity of hot flashes with gabapentin, although data across the studies were too heterogeneous to provide a reliable summary effect. Clusterings of dizziness/unsteadiness and fatigue/somnolence were the most frequently reported adverse events associated with gabapentin and resulted in a higher dropout rate due to adverse events in the gabapentin-treated patients than in the controls. More studies are needed to consolidate the outcomes and elucidate useful details regarding this treatment. PMID- 19302897 TI - The relationship between reduction in low-density lipoprotein cholesterol by statins and reduction in risk of cardiovascular outcomes: an updated meta analysis. AB - BACKGROUND: In 2005, the Cholesterol Treatment Trialists' Collaboration (CTTC) quantified the relationship between reduction in low-density lipoprotein cholesterol (LDL-C) achieved by statin treatment and reduction in cardiovascular risk. Since this publication, several large statin trials have been reported. OBJECTIVE: The objective of our analysis was to extend the CTTC results by including active-controlled trials and other trials published since 2005. METHODS: A literature search in English (1966-December 2008) was undertaken of MEDLINE, EMBASE, Derwent drug file databases, and the Cochrane library using standard MESH terms (cardiovascular disease, death, fatal outcome, pravastatin, simvastatin, atorvastatin, rosuvastatin, fluvastatin, lovastatin, and hydroxymethylglutaryl coenzyme A reductase inhibitors) to identify randomized trials of statins (placebo controlled, active controlled, or usual care) that reported clinical outcomes, enrolled >1000 subjects, and followed them up for > or =1 year. Random effects meta-regression was used to analyze the relationship between absolute changes in LDL-C and risk for cardiovascular events. RESULTS: Twenty-five trials were included in a primary analysis involving 155,613 subjects, 6321 vascular deaths, 23,791 major vascular events, 11,357 major coronary events, and 4717 strokes. For every 25-mg/dL (0.65-mmol/L) reduction in LDL-C, the relative risk (95% CI) for various cardiovascular outcomes was as follows: vascular mortality, 0.89 (0.87-0.92); major vascular events, 0.86 (0.84 0.88); major coronary events, 0.84 (0.82-0.86); and stroke, 0.90 (0.86-0.94). CONCLUSIONS: Based on meta-regression analysis of these trials, there was a significant positive relationship between reduction in LDL-C and reduction in the risk for major cardiovascular events. These results support and extend the findings of the CTTC. PMID- 19302898 TI - Colesevelam hydrochloride for the treatment of type 2 diabetes mellitus. AB - BACKGROUND: Colesevelam hydrochloride is a bile acid sequestrant approved in January 2008 by the US Food and Drug Administration (FDA) for the treatment of adult patients with type 2 diabetes mellitus (DM) in combination with a sulfonylurea, metformin, and/or insulin therapy. OBJECTIVE: The purpose of this article was to review the pharmacology, pharmacokinetics, efficacy, adverse effects and tolerability, drug-drug interactions, contraindications/precautions, dosage and administration, pharmacoeconomics, and the overall role of colesevelam in the management of adult patients with type 2 DM. METHODS: A literature search using MEDLINE (1966-October 27, 2008), PubMed (1950-October 27, 2008), Science Direct (1994-October 27, 2008), Web of Science (1980-October 27, 2008), American Diabetes Association Scientific Abstracts (2004-2008), and International Pharmaceutical Abstracts (1970-October 27, 2008) was performed using the term colesevelam. English-language, original research and review articles were examined, and citations from these articles were assessed. Manufacturer prescribing information and the FDA review of the new drug application for colesevelam were also examined. RESULTS: Colesevelam is a hydrophilic, water insoluble polymer, with negligible absorption and systemic distribution, that is excreted primarily in the feces. Through a mechanism still under investigation, colesevelam effectively lowers glycosylated hemoglobin (HbA(1c)) when used in combination with a sulfonylurea, metformin, and/or insulin therapy. Three completed, published Phase III clinical trials investigating colesevelam for the treatment of type 2 DM were evaluated for information, data, and conclusions. At dosing of 1.875 g BID or 3.75 g once daily in combination with one of the aforementioned agents versus placebo, reductions in HbA(1c) in all 3 Phase III clinical trials of colesevelam ranged from 0.5% to 0.7% (P < 0.02). In clinical trials, colesevelam was well tolerated, with hypoglycemia occurring in approximately 3% of studied patients. CONCLUSIONS: When used in combination with a sulfonylurea, metformin, and/or insulin therapy, colesevelam has been reported to significantly reduce HbA(1c) in adult patients with type 2 DM. Colesevelam's role in the management of type 2 DM remains undefined, however; further investigation into its mechanism of action and long-term efficacy and safety should be performed. PMID- 19302899 TI - Efficacy and tolerability of tapentadol immediate release and oxycodone HCl immediate release in patients awaiting primary joint replacement surgery for end stage joint disease: a 10-day, phase III, randomized, double-blind, active- and placebo-controlled study. AB - OBJECTIVES: The primary objective of this study was to assess the efficacy and tolerability of tapentadol immediate release (IR) in patients who were candidates for joint replacement surgery due to end-stage joint disease. A secondary objective was to compare tapentadol IR with oxycodone HCl IR with respect to efficacy and prespecified tolerability end points. METHODS: This 10-day, Phase III, randomized, double-blind, active- and placebo-controlled study compared the efficacy and tolerability of tapentadol IR, oxycodone HCl IR, and placebo in patients with uncontrolled osteoarthritis pain who were candidates for primary replacement of the hip or knee as a result of end-stage degenerative joint disease. Patients received tapentadol IR 50 mg, tapentadol IR 75 mg, oxycodone HCl IR 10 mg, or placebo every 4 to 6 hours during waking hours. The primary end point was the sum of pain intensity difference (SPID) over 5 days. Secondary efficacy end points included 2- and 10-day SPID; 2-, 5-, and 10-day total pain relief (TOTPAR); and the sum of total pain relief and pain intensity difference (SPRID). Prespecified noninferiority comparisons with oxycodone HCl IR were performed with respect to efficacy (based on 5-day SPID) and tolerability (based on incidence of the reported adverse events (AEs) of nausea and/or vomiting and constipation). RESULTS: Of 666 patients originally enrolled, 659 were included in the efficacy analysis (51% male; 91% white; mean age, 61.2 years; mean weight, 97 kg). Five-day SPID was significantly lower in those treated with tapentadol IR (tapentadol IR 50 mg: least squares mean difference [LSMD] = 101.2 [95% CI, 54.58 147.89]; tapentadol IR 75 mg: LSMD = 97.5 [95% CI, 51.81-143.26]) or oxycodone HCl IR (LSMD = 111.9 [95% CI, 66.49-157.38]) (all, P < 0.001). Tapentadol IR 50 and 75 mg and oxycodone HCl IR 10 mg were associated with significant reductions in pain intensity compared with placebo, based on 2- and 10-day SPID and 2-, 5-, and 10-day TOTPAR and SPRID (all, P < 0.001). The efficacy of tapentadol IR 50 and 75 mg was noninferior to that of oxycodone HCl IR 10 mg; however, the incidence of selected gastrointestinal AEs (nausea, vomiting, and constipation) was significantly lower for both doses of tapentadol IR compared with oxycodone HCl IR 10 mg (nominal P < 0.001). The odds ratios for nausea and/or vomiting for tapentadol IR 50 and 75 mg relative to oxycodone HCl IR 10 mg were 0.21 (95% CI, 0.128-0.339) and 0.32 (95% CI, 0.204-0.501), respectively; for constipation, the corresponding odds ratios were 0.13 (95% CI, 0.057-0.302) and 0.20 (95% CI, 0.098 0.398). Rates of treatment discontinuation were 18% (28/157) in the tapentadol IR 50-mg group, 26% (43/168) in the tapentadol IR 75-mg group, 35% (60/172) in the oxycodone HCl IR 10-mg group, and 10% (17/169) in the placebo group. In a post hoc analysis, tapentadol IR 50 mg was associated with a significantly lower incidence of treatment discontinuation than was oxycodone HCl IR 10 mg (P < 0.001). CONCLUSIONS: In these patients with uncontrolled osteoarthritis pain who were awaiting joint replacement surgery, tapentadol IR 50 and 75 mg were associated with analgesia that was noninferior to that provided by oxycodone HCl IR 10 mg. Tapentadol treatment was associated with improved gastrointestinal tolerability. PMID- 19302900 TI - Comparison of the effects of once-monthly versus once-daily risedronate in postmenopausal osteoporosis: a phase II, 6-month, multicenter, randomized, double blind, active-controlled, dose-ranging study. AB - BACKGROUND: Risedronate 5 mg/d is approved by the US Food and Drug Administration for the treatment and prevention of postmenopausal osteoporosis. Once-monthly dosing options might increase treatment compliance and persistence. OBJECTIVE: The aim of this study was to compare the tolerability and efficacy of 3 once monthly risedronate dosing regimens with those of risedronate 5 mg/d. METHODS: This Phase II, 6-month, randomized, double-blind, active-controlled, dose-ranging study was conducted at 13 clinical research centers and hospitals in Croatia, Poland, Canada, and the United States between April 2004 and June 2005. Post menopausal women aged 50 to 85 years with a lumbar spine T-score <-2.0 were randomly assigned to 1 of 4 treatment groups: risedronate 100, 150, or 200 mg/mo or 5 mg/d (active control), administered PO for 6 months. Evaluation of tolerability, the primary study objective, was based on adverse-event (AE) profiles and clinical laboratory values. Efficacy evaluation, a secondary objective, was a noninferiority comparison of the changes from baseline in bone mineral density (BMD) and bone turnover markers (BTMs). RESULTS: Of 370 patients randomized (91, 88, 88, and 103 patients in the risedronate 100-, 150-, and 200 mg/mo and 5-mg/d groups, respectively), 57% were > or =65 years of age and 99% were white; 316 patients (85.4%) completed the study. Completion rates were not significantly different across treatment groups, nor were reasons for discontinuation. Between-group differences in the incidences of treatment emergent AEs, serious AEs, and upper gastrointestinal (GI) AEs were nonsignificant. Overall, 6 (7%), 14 (16%), 6 (7%), and 9 patients (9%) withdrew because of AEs in the 100-, 150-, and 200-mg/mo and 5 mg/d groups, respectively. GI disorders were the AEs that most frequently led to study withdrawal (5 [5.5%], 7 [8.0%], 4 [4.5%], and 6 [5.8%]). No trends were observed in the nature or frequency of other AEs causing withdrawal. All serious AEs were considered unrelated to treatment, with the exception of erosive esophagitis in 1 patient (1%) who received the 5-mg/d dose. Mean percentage increases in BMD were 2.10%, 2.99%, and 3.38% with risedronate 100, 150, and 200 mg/mo, respectively, versus 3.05% with 5 mg/d. At the 2 higher monthly doses, the changes from baseline in BMD were not significantly different from those in the 5-mg/d group. Mean BTM values were decreased significantly from baseline in all 4 treatment groups, and the changes from baseline at 6 months at the 2 higher monthly doses were not significantly different from those at 5 mg/d. CONCLUSIONS: Overall, in this study, the safety profiles of risedronate 100, 150, and 200 mg/mo were not different from that of risedronate 5 mg/d. Changes in efficacy measures in the monthly treatment groups were considered dose related and were not significantly different between the 5-mg/d group and the 150- and 200-mg/mo groups; similarity was greatest with 150 mg/mo. PMID- 19302901 TI - Effects of oral posaconazole on the pharmacokinetic properties of oral and intravenous midazolam: a phase I, randomized, open-label, crossover study in healthy volunteers. AB - BACKGROUND: Like itraconazole and ketoconazole, posaconazole, a broad-spectrum oral triazole antifungal, inhibits the activity of the cytochrome P450 (CYP) isozyme 3A4. Midazolam, a short-acting benzodiazepine, is metabolized by CYP3A4. Potential drug interactions can be expected in patients who are concurrently receiving inhibitors and substrates of CYP3A4 (eg, ketoconazole, posaconazole) and benzodiazepines (eg, midazolam). Because of the potential for drug interactions, it is important to determine the effects of posaconazole on the pharmacokinetic properties of midazolam. OBJECTIVE: The aim of this study was to compare the effects of oral administration of posaconazole versus ketoconazole on the pharmacokinetic properties of orally and intravenously administered midazolam. METHODS: This Phase I, randomized, open-label, crossover study was conducted at Swiss Pharma Contract Ltd., Allschwil, Switzerland. Healthy volunteers were randomly assigned to 1 of 2 treatment arms. Arm 1 received posaconazole 200 mg BID for 7 days, posaconazole 400 mg BID for 7 days, no drugs during a 28-day washout, and ketoconazole 400 mg once daily for 7 days. Arm 2 received posaconazole and ketoconazole in the reverse order, with a 28-day washout between treatments. An oral/IV midazolam sequence (oral midazolam 2 mg and IV midazolam 0.4 mg) was administered on days -2/-1, 6/7, 13/14 (arm 1), 36/17 (arm 2), 43/44, and 50/51 in both treatment arms. Blood samples were collected up to 24 hours after midazolam administration. Pharmacokinetic parameters, including C(max), C(min) (before azole administration), terminal phase t(1/2) (t(1/2z)), and AUC to final measurable sampling time (AUC(tf)), were calculated using noncompartmental methods, and drug interactions were evaluated using analysis of variance. Adverse events were collected using physical examination, including vital sign measurements; clinical laboratory analysis; electrocardiography; and direct questioning at predefined time points throughout the study to assess tolerability. RESULTS: A total of 12 subjects were enrolled (11 men, 1 woman; all white; mean age, 42.8 years [range, 28-53 years]; mean weight, 80.6 kg; and mean body mass index, 25.6 kg/m(2)). All of the subjects completed the study. Based on point estimates of logarithm-transformed data, posaconazole 200 and 400 mg BID were associated with significant increases in midazolam C(max) (up to 1.3- and 2.4-fold) and AUC(tf) values (up to 4.6- and 6.2 fold), respectively. Ketoconazole 400 mg once daily was associated with significantly increased midazolam C(max) and AUC(tf) (up to 2.8- and 8.2-fold, respectively). When midazolam was concurrently administered with either azole, t(1/2z) was prolonged. Seven of 12 (58%) subjects reported > or =1 adverse event during the study (5 with posaconazole alone and 4 with posaconazole + midazolam). The most common adverse events were diarrhea (3 subjects [25%] with posaconazole alone, 2 [17%] with ketoconazole alone, and 1 [8%] with posaconazole + midazolam) and flatulence (1 [8%] with posaconazole alone and 1 [8%] with midazolam alone). CONCLUSIONS: The results from this study in a small, all-white population of healthy volunteers suggest that posaconazole was a potent inhibitor of CYP3A4, but to a lesser extent than was ketoconazole. Monitoring patients for adverse events, the need for dose adjustments, or both during coadministration with posaconazole may be warranted in patients being treated with benzodiazepines that are predominantly metabolized through CYP3A4 (eg, midazolam). PMID- 19302902 TI - Effects of an alternative cefepime dosing strategy in pulmonary and bloodstream infections caused by Enterobacter spp, Citrobacter freundii, and Pseudomonas aeruginosa: a single-center, open-label, prospective, observational study. AB - BACKGROUND: Various dosing strategies for cefepime have been developed in an effort to maximize pharmacodynamic exposure of this agent against gram-negative infections. An assessment of cefepime dosing strategies is warranted given recent reports of poorer treatment outcomes associated with cefepime compared with other antibiotics, particularly in patients infected with gram-negative organisms with elevated MICs. OBJECTIVES: The aims of this study were to compare the efficacy of cefepime IV at a dose of 1 g q8h (adjusted based on renal function) with those of other appropriate antimicrobials in the treatment of gramnegative pulmonary and bloodstream infections and to identify risk factors for treatment failure. METHODS: This single-center, open-label, prospective, observational study was conducted at a tertiary care center (Barnes-Jewish Hospital, St. Louis, Missouri). Isolates from infections in adult patients with bacteremia or pulmonary infection caused by Pseudomonas aeruginosa, Enterobacter aerogenes, Enterobacter cloacae, or Citrobacter freundii were assessed in a noninterventional manner. Infections were identified using an electronic notification system. Patients receiving appropriate monotherapy against the studied isolate within 24 hours of culture attainment were stratified into 1 of 3 cohorts according to treatment outcome, as follows: treatment success (resolution of initial fever or elevated white blood cell count to normal values plus the presence of repeat negative cultures from the initial site or below the quantitative definition for infection), improvement (treatment success without repeat negative cultures), or treatment failure (persistent or repeat positive cultures for the original organism at the infected site despite appropriate and adequate antimicrobial therapy, lack of resolution in fever or leukocytosis, switch to an alternative antibiotic, or the addition of another antibiotic with gram-negative coverage after > or =3 days of the initial regimen, relapse of infection within 14 days, or mortality attributable to the index infection). Multivariate regression analysis was used to examine risk factors associated with treatment failure. RESULTS: Data from 120 patients (56.7% male; mean age, 62.2 years) were analyzed. Treatment failure occurred in 48.6% (36/74) of patients who received cefepime versus 32.6% (15/46) of those who received other antibiotics; this difference was not statistically significant. The proportion of patients with markers of increased severity of illness (intensive care unit [P = 0.005] and mechanical ventilation [P = 0.002]) was significantly greater in the cefepime group compared with the group that received other antibiotics. Multivariate logistic regression identified infection with Pseudomonas aeruginosa (adjusted odds ratio [AOR], 1.40 [95% CI, 1.01-2.00]) and mechanical ventilation (AOR, 7.08 [95% CI, 1.80-31.3]) as being associated with treatment failure in patients who received cefepime. Mechanical ventilation (AOR, 3.97 [95% CI, 1.47-11.1]) and neutropenia (AOR, 5.26 [95% CI, 1.28-20.0]) were independent predictors of treatment failure among all patients studied. CONCLUSIONS: Based on these results in this small cohort, the efficacy of this cefepime dosing strategy (1 g q8h) appeared to be similar to that of other antimicrobials. PMID- 19302903 TI - Pharmacokinetic properties of S-adenosylmethionine after oral and intravenous administration of its tosylate disulfate salt: a multiple-dose, open-label, parallel-group study in healthy Chinese volunteers. AB - BACKGROUND: S-adenosylmethionine (SAMe) is an endogenous molecule that plays an important role in cellular metabolism. Despite being widely used as a dietary supplement with claimed benefits for numerous conditions, there is little information about the pharmacokinetic properties of exogenous SAMe. OBJECTIVES: One aim of this study was to characterize the pharmacokinetic properties of SAMe after administration of single and multiple doses of orally and intravenously administered SAMe tosylate disulfate (STD) in healthy male and female Chinese volunteers. Because men have higher erythrocyte levels of endogenous SAMe than do women, we also assessed the effects of sex on the disposition of SAMe. METHODS: A simple and sensitive assay for SAMe based on liquid chromatography-mass spectrometry using selected-ion monitoring of analyte and acyclovir as internal standard was developed and validated. The assay was used to study the pharmacokinetic properties of SAMe. STD was administered as single and multiple doses of enteric-coated tablets and IV infusion of STD to groups of healthy native Chinese volunteers. After an overnight fast, male and female Chinese volunteers were assigned to receive STD 1000 mg for 5 days, either in enteric coated tablet formulation or as a 250-mL IV infusion. Blood samples were collected 24 hours after the first and last dose and used for determining plasma SAMe concentrations and pharmacokinetic parameters. For the oral formulation, SAMe concentrations were corrected for concentrations of endogenous SAMe. Pharmacokinetic parameters were calculated for men and women separately and for the total group of volunteers. Adverse events were monitored using a physician during blood collection and by spontaneous reporting. RESULTS: Twenty healthy volunteers were enrolled (oral formulation: 5 men, 5 women; mean [SD] age, 24.1 [4.7] years [range, 21-37 years]; mean [SD] weight, 59.9 [4.8] kg [range, 54-70 kg]; IV formulation: 5 men, 5 women; mean [SD] age, 22.6 [1.8] years [range, 21 27 years]; mean [SD] weight, 59.5 [5.4] kg [range, 53-67 kg]). None of the between-sex differences in SAMe pharmacokinetic properties were significant. The (mean [SD]) pharmacokinetic properties of singledose oral SAMe in men and women, respectively, were as follows: C(max), 2.37 (1.58) and 2.50 (1.83) micromol/L; T(max), 5.40 (1.14) and 5.20 (1.48) hours; AUC(0-24), 8.56 (5.16) and 10.3 (8.0) micromol/L/h; and t(1/2beta), 6.06 (1.80) and 6.28 (2.60) hours. Corresponding values with the single-dose IV formulation were: C(max), 127 (49) and 211 (94) micromol/L; T(max), 1.90 (0.22) and 1.60 (0.22) hours; AUC(0-24), 329 (84) and 480 (176) micromol/L/h; and t(1/2beta), 4.34 (0.57) and 3.83 (0.78) hours. The single-dose oral:IV ratios of AUC(0-24) in men and women, respectively, were 2.60% and 2.14% (degrees of fluctuation: 4.96 [1.77] and 9.49 [0.91]). The pharmacokinetic properties of multiple-dose oral and IV SAMe were not significantly different from those with single-dose administration. None of the volunteers reported any adverse events during the study. CONCLUSIONS: In this small study in healthy Chinese volunteers, there were no significant differences in the pharmacokinetic parameters of SAMe between men and women or between single and multiple-dose administration of STD 1000 mg administered orally or intravenously. No evidence of accumulation of SAMe in plasma was found on multiple dosing. Both enteric-coated tablets and the IV infusion were well tolerated in these volunteers. PMID- 19302904 TI - Evaluation of a compliance device in a subgroup of adult patients receiving specific immunotherapy with grass allergen tablets (GRAZAX) in a randomized, open label, controlled study: an a priori subgroup analysis. AB - OBJECTIVES: This a priori subgroup analysis was conducted to assess patients' experience with a compliance device for the administration of sublingual specific immunotherapy for grass pollen-induced rhinoconjunctivitis. METHODS: The present paper reports the results of a subgroup analysis of a multicenter, randomized, controlled, open-label European study in which adults with grass pollen-induced rhinoconjunctivitis received once-daily treatment with grass allergen tablets with or without a device to aid compliance. Treatment was given approximately 6 to 12 weeks before the grass pollen season, during the season (approximately 8-10 weeks), and for up to 2 weeks after the end of the season. In the subgroup analysis, patients from Germany and the Netherlands who had been randomized to receive the compliance device were asked to complete a brief questionnaire at the final study visit. The questionnaire included 6 items concerning patients' use of the device and whether it helped them remember to take their tablets. RESULTS: Of the 91 patients from Germany and the Netherlands who were randomized to use the compliance device along with grass allergen tablets, 71 returned the questionnaire. Among responders, 58 of 71 (82%) reported using the device sometimes or always, 50 of 63 (79%) found the device easy to use, 32 of 69 (46%) found that the device made it easier to remember to take tablets, and 43 of 71 (61%) indicated that they would consider using the device again. CONCLUSION: Most patients in this subgroup analysis used the compliance device as a medication reminder and rated it easy to use. PMID- 19302905 TI - Pharmacokinetics of 14C-radioactivity after oral intake of a single dose of 14C labeled fampridine (4-aminopyridine) in healthy volunteers. AB - BACKGROUND: Fampridine (4-aminopyridine) is a potassium channel blocker that has been evaluated as a treatment for patients with spinal cord injury and multiple sclerosis. OBJECTIVE: The purpose of this study was to determine the pharmacokinetics of a single dose of an orally administered solution of (14)C labeled fampridine in healthy volunteers. METHODS: In this open-label, single dose study conducted in an inpatient setting, healthy adult men were administered an oral solution containing 15 mg of (14)C-labeled fampridine (100 microCi) in a fasted state. In addition to blood sampling for analysis of plasma (14)C radioactivity at prescribed intervals over 7 days, all urine and feces were collected for analysis of drug recovery and disposition. Urine samples were also analyzed for metabolic profiling. Plasma pharmacokinetic parameters of the (14)C radiolabeled drug were determined using standard liquid-scintillation techniques. Recovery was calculated to provide the total amount of radioactivity excreted as a proportion of the original dose. Nonhydrolyzed and hydrolyzed urine extracts were analyzed for radioactivity and metabolites using reverse-phase, isocratic high-performance liquid chromatography with spectrophotometric and radioactive detection. Tolerability was assessed through evaluation of vital signs, hematologic and other laboratory parameters, and electrocardiography. RESULTS: The 4 white male subjects had a mean (SD) age of 21 (2) years. No clinically significant abnormalities in vital signs, clinical chemistry, hematology, urinalysis, or electrocardiography were observed either before or during the study. Peak plasma radioactivity was reached at 1 hour after dosing, with a median concentration of 72.9 ng x mL(-1). There was complete disappearance of radioactivity by 24 hours (limit of quantitation, 400 disintegrations/min per peak), and the calculated median t(1/2) was 3.14 hours. Total cumulative recovery of (14)C-radioactivity was 96.36%, with only 0.51% of drug recovered in feces. On chromatography, 2 metabolites accounted for a low proportion of total urinary radioactivity (3% and 6% of total radioactivity in the interval from 0 to 4 hours after dosing; 17% and 9% in the interval from 8 to 12 hours after dosing). Three subjects reported mild and transient dizziness occurring 1 half-hour after dosing; this was considered possibly related to the study drug. CONCLUSION: Fampridine administered as an oral solution was rapidly absorbed and was nearly completely and rapidly eliminated as unchanged drug via urinary excretion, suggesting that it is unlikely to undergo substantial metabolic transformation. PMID- 19302906 TI - Impact of illegal trade on the quality of epoetin alfa in Thailand. AB - BACKGROUND: Reports from the World Health Organization have suggested that counterfeit medicines pose a serious problem in developing countries. An investigation of anti-erythropoietin antibody-mediated pure red cell aplasia in Thailand found evidence of drug smuggling, which may have serious safety implications. OBJECTIVE: This study assessed the authenticity and quality of epoetin alfa samples in Thailand. METHODS: Samples of epoetin alfa-prefilled syringes were collected from the pharmacies at 2 major hospitals (62 samples), 8 retail pharmacies (41 samples), and Thai authorities (30 samples confiscated from smugglers at 2 airports, and 6 samples from a condominium used by smugglers). These samples were tested against the European Union Pharmacopeia specifications for aggregate content in epoetins of <2%. The integrity of epoetin alfa distribution channels, coldchain processes (maintenance at 2 degrees C-8 degrees C), primary and secondary packaging components (eg, batch number, expiration date, appearance, letter size), and company's confidential features (eg, nature of the ink, type and quality of the paper, other covered features) were also investigated. The main outcome measures were protein aggregate content, determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting; and isoform distribution, assessed by isoelectric focusing and Western blotting. RESULTS: Epoetin alfa samples obtained from the company's cold chain and authorized distribution channels met all quality standards, as did all epoetin alfa samples obtained from the hospital pharmacies. However, evidence showed that some samples were being smuggled or sold illegally through certain unauthorized retail pharmacies. The epoetin alfa samples obtained from both airports and the condominium were stored improperly at room temperature. Aggregate levels exceeded the specification of <2% in 11 samples from 2 of the retail pharmacies (range, 1.2%-3.1%), 15 samples from the Dongmuang Airport (range, 2.2%-17.0%), and all 6 samples from the condominium (range, 10.5%-19.8%). All samples from the 2 hospitals, 8 retail pharmacies, and Suvarnabhumi Airport had the authentic 6 isoform bands. Samples from Dongmuang Airport and the condominium appeared to have the 6 characteristic bands, but positive confirmation was difficult because of band smearing caused by a high level of aggregates. All features of primary and secondary packaging were found to be authentic. CONCLUSIONS: This investigation found evidence that some epoetin alfa samples were smuggled into Thailand without proper cold chain, contained high levels of protein aggregates, and were sold illegally through certain retail pharmacies. The Thai authorities have intervened to stop such unauthorized products from reaching patients. Strenuous efforts must be made to prevent illegal cross-border smuggling of biopharmaceuticals without proper cold chain because of the serious safety implications for patients in developing countries. PMID- 19302907 TI - Oxymorphone extended release for the treatment of chronic low back pain: a retrospective pooled analysis of enriched-enrollment clinical trial data stratified according to age, sex, and prior opioid use. AB - OBJECTIVE: This study assessed the potential effects of age, sex, and prior opioid use on the response to oxymorphone extended release (ER) in patients with moderate to severe chronic low back pain. METHODS: Combined data from 2 placebo controlled clinical trials with an enriched-enrollment, randomized-withdrawal design were analyzed. In patients aged > or =18 years with chronic low back pain, the dose of oxymorphone ER was titrated to a stable, tolerable, effective dose. Patients who completed titration were randomly assigned to a 12-week double-blind study period with oxymorphone ER or placebo. Oxymorphone immediate release 5 mg was permitted q4-6h, as needed for rescue medication or withdrawal symptoms, for 4 days after randomization and restricted to 10 mg/d thereafter. Pain intensity (100-mm visual analog scale [VAS]; 0 = no pain to 100 = worst pain imaginable) and time to study discontinuation due to lack of efficacy were compared with stratification by age (<65 vs > or =65 years), sex, and prior opioid use. Adverse events were categorized by severity and relation to study medication. RESULTS: Of 575 patients, 348 completed titration and 347 entered the double-blind study phase. There were no significant between-group differences in demographic variables, except that the mean age in the oxymorphone ER group was significantly higher (P = 0.04), and the proportion of men was significantly lower (P = 0.01). There was no significant age difference between the oxymorphone ER and placebo groups stratified by age (<65 vs > or =65 years). Fewer patients aged > or =65 years versus <65 years completed titration (45.0% [36/80] vs 63.0% [312/495]; P = 0.002). The least-squares mean (SEM) differences in VAS pain scores between the oxymorphone ER (n = 174) and placebo (n = 169) groups were significant at each postbaseline assessment (P < 0.001) and at study completion (12.3 [2.8] mm; P < 0.001) and was not significantly affected by age, sex, or prior opioid use. Age and sex had no significant influence on adverse events or discontinuations due to lack of efficacy. More discontinuations due to lack of efficacy occurred among patients in the placebo group (hazard ratio, 5.01; P < 0.001) and among opioid experienced patients. The latter effect was limited to opioid-experienced patients who received placebo. The rates of discontinuation due to lack of efficacy were similar between oxymorphone ER-treated opioid-naive and opioid experienced patients (11.4% vs 11.6%). The proportion of patients who experienced opioid-related adverse events was significantly greater in the oxymorphone ER group compared with the placebo group (25.7% vs 16.3%; P = 0.03). The most frequent treatment-emergent adverse events in the oxymorphone ER group were nausea (8.0%), constipation (6.3%), vomiting (4.6%), and diarrhea (4.0%); in the placebo group were nausea (5.8%), diarrhea (4.7%), and increased sweating (2.3%). CONCLUSION: In the enriched population of patients who successfully titrated to oxymorphone ER, oxymorphone ER was effective and generally well tolerated, independent of patients' age, sex, or previous opioid use. PMID- 19302908 TI - Evolution of hepatitis B virus polymerase mutations in a patient with HBeAg positive chronic hepatitis B virus treated with sequential monotherapy and add-on nucleoside/nucleotide analogues. AB - BACKGROUND: Nucleoside/nucleotide analogues are a fundamental tool for the treatment of chronic hepatitis B virus (HBV). Sequential anti-HBV treatment might lead to the selection of mutations. OBJECTIVE: This report aimed to analyze the genetic evolution of the reverse-transcriptase (RT) gene of viral quasispecies in a patient with hepatitis B e antigen (HBeAg)-positive chronic HBV who received, sequentially, lamivudine (LAM), adefovir dipivoxil (ADV), and ADV + telbivudine (LDT) combination treatment over a total of 108 weeks. METHODS: A 20-year-old Chinese man presented to Huashan Hospital, Fudan University, Shanghai, People's Republic of China, with hepatitis B surface antigen-positive and HBeAg-positive chronic HBV and was sequentially treated with LAM 100 mg/d for 18weeks,ADV 10mg/d for 68weeks, and ADV 10mg/d + LDT 600 mg/d combination treatment for 22 weeks. Compliance was monitored every 4 weeks using a pill count. For genotypic analysis, the RT region of the polymerase gene from the serum of this patient was amplified, cloned, and sequenced. Fifty clones with HBV insert were selected for sequencing at weeks 0 (baseline), 18, 22, 60, 70, 86, and 108. RESULTS: The rtM204V/L LAM-resistance mutation was detected in 4.4% (2/45) of clones prior to LAM treatment. At week 18 during LAM treatment, the rtM204I mutation became predominant, being present in 79.5% (35/44) of clones. The rtM204I mutation was associated with compensatory mutations (rtL180M and rtT184L). A total of 9.1% (4/44) of the clones harbored the rtL180M + rtT184L + rtM204I mutations. Two new mutations, rtL229V and rtV191I, were detected in 75.0% (33/44) and 11.4% (5/44) of clones, respectively. At week 22 during ADV treatment, LAM-resistance mutations (rtL180M, rtT184L, rtM204I, rtV191I, and rtL229V) were not detected. At week 86 during ADV therapy, the rtN236T ADV-resistance mutation was detected in 58.8% (20/34) of clones. A total of 20.6% (7/34) of the clones harbored the rtK212T + rtM250L mutation, and rtA181V was found in 2.9% (1/34) of the clones. At week 108, after the patient had been receiving ADV + LDT combination therapy for 22 weeks, rtS202G and rtI269T had emerged, representing 28.9% (13/45) and 8.9% (4/45), respectively, of the viral population during ADV + LDT combination treatment. We also detected several polymorphic sites,including rtF221Y, rtS223A, rtI224V, rtN238H, rtL267Q, and rtQ271M, during the sequential treatment. After 22 weeks of combination treatment, HBV DNA count was decreased to less than the lower limit of quantitation (<200 copies/mL). CONCLUSIONS: This report identified HBV mutations that escaped the antiviral pressure of LAM, ADV, and ADV + LDT in this patient and provided insight into the process of mutation selection through genotypic analysis during antiviral treatment. Mutations selected under sequential treatments of LAM, ADV, and ADV + LDT can lead to a series of compensatory mutations, which partially restore the level of viral replication. ADV administered in combination with LDT appeared to be effective in this selected case with clinical or virologic resistance to sequential treatment with LAM and ADV. PMID- 19302909 TI - Requirements for publication of bioequivalence pharmacokinetic studies in Clinical Therapeutics. PMID- 19302910 TI - Pharmacokinetic, pharmacodynamic, efficacy, and safety data from two randomized, double-blind studies in patients with asthma and an in vitro study comparing two dry-powder inhalers delivering a combination of salmeterol 50 microg and fluticasone propionate 250 microg: implications for establishing bioequivalence of inhaled products. AB - BACKGROUND: The use of dry-powder inhalers (DPIs) to administer respiratory medicines is increasing, and new DPIs are likely to be developed because of expiring patents. However, there is considerable debate concerning the extent to which DPIs are interchangeable without altering disease control or the safety profile of the treatment. OBJECTIVE: This study was designed to compare the pharmacokinetic (PK), pharmacodynamic (PD), efficacy, and safety data for 2 DPIs delivering a combination of salmeterol 50 microg plus fluticasone propionate (FP) 250 microg (SFC 50/250) to investigate assumptions of bioequivalence. METHODS: Three studies compared SFC 50/250 delivery using a reservoir powder inhalation device (RPID) and a Diskus multiple-dose inhaler: an in vitro assessment of fine particle-mass (FPM) profiles of the emitted doses; a PK/PD study of SFC 50/250 administered in two 14-day crossover treatment periods to 22 adults with moderate, persistent asthma to determine the equivalence of the RPID and Diskus inhaler in terms of drug delivery and systemic exposure; and a 12-week clinical efficacy and safety study of SFC 50/250 in 270 patients > or =12 years of age with moderate, persistent asthma to assess the equivalence of the RPID and Diskus inhaler based on peak expiratory flow (PEF) rates. FPM was summed from the quantity of active pharmaceutical ingredient deposited on stages 1 to 5 of a cascade impactor, representing an aerodynamic particle size range of 0.8 to 6.2 microm. Systemic exposure to SFC 50/250 was declared no greater with RPID than with the Diskus inhaler if the upper limit of the 90% CI for the ratio of FP AUC for the 2 devices was below the upper limit of the equivalence range (ie, <1.25). Adverse events, clinical laboratory test results, and vital signs were recorded throughout the 2 clinical studies. RESULTS: In vitro, mean FPM values for the RPID and Diskus inhaler, respectively, were 13.1 and 12.8 microg/dose for salmeterol (P = NS) and 66.8 and 66.2 microg/dose for FP (P = NS). The only notable differences were mean FP for particle sizes 2.3 to 3.2 microm (21.4 microg/dose for RPID, 25.6 microg/dose for Diskus) and for sizes 4.0 to 6.2 microm (17.3 microg/dose for RPID, 11.7 microg/dose for Diskus). In the PK/PD study, there were 22 patients (16 men and 6 women), most (86%) of whom were white. Mean (SD) age was 26.0 (5.0) years (range, 19-35 years), and mean (SD) weight was 67.3 (8.9) kg. The 2 inhalers did not meet the criteria for declaring bioequivalence: estimated ratios (RPID:Diskus) were 2.00 (90% CI, 1.56 to 2.55) for FP AUC up to the time point of next dosing and 1.92 (90% CI, 1.64 to 2.25) for salmeterol maximum observed plasma concentration at the end of the dosing interval (at steady state). Urine cortisol (0-24 hours) was significantly lower for the RPID than for the Diskus inhaler (ratio, 0.74 [95% CI, 0.57 to 0.96]; P = 0.026); no significant difference in plasma cortisol was noted between the 2 inhalers (ratio, 0.85 [95% CI, 0.7 to 1.04]). A small but statistically significant increase in maximum heart rate (5 beats/min) was noted in the RPID group (ratio, 1.05 [95% CI, 1.01 to 1.10]; P = 0.029). No notable differences in other PD end points were observed. Drug-related adverse events occurred in both groups (2 [dysphagia and tremor] in the RPID group and 3 [2 cases of dysphonia, 1 case of mucous-membrane irritation] in the Diskus group). There were 270 patients (136 females, 134 males) in the clinical efficacy and safety study, most (94%) of whom were white; mean (SD) age was 37.2 (17.0) years (range, 11-77 years) in the RPID group and 35.4 (17.2) years (range, 12-77 years) in the Diskus group. The RPID and the Diskus inhaler met the predefined equivalence criteria (+/-15 L/min) in terms of mean change in morning PEF from baseline: 3.9 L/min (95% CI, -3.1 to 11.0). The 2 SFC 50/250 inhalers were well tolerated; the most frequently reported adverse event was bronchitis, reported by 12% of the patients in the RPID group and 9% of those in the Diskus group. The only serious adverse event, which occurred in the RPID group and was related to bronchial infection, was considered unrelated to treatment. CONCLUSIONS: In vitro particle size distribution data were potentially superimposable for the RPID and the Diskus inhaler. The 2 devices were considered to be clinically equivalent in terms of mean morning PEF but were not considered equivalent in terms of PK systemic exposure. The 2 SFC 50/250 inhalers were well tolerated and had comparable safety profiles; no serious adverse events were attributed to the study product. PMID- 19302911 TI - Pharmacokinetics, bioequivalence, tolerability, and effects on platelet counts of two formulations of anagrelide in healthy volunteers and patients with thrombocythemia associated with chronic myeloproliferation. AB - BACKGROUND: Anagrelide hydrochloride is an anti-thrombotic agent indicated for the treatment of essential thrombocythemia (ET). In various previously published clinical trials of 2 branded formulations of anagrelide in patients with ET at high risk for thrombohemorrhagic events, the rates of adverse events and discontinuation were strikingly divergent between brands. Because the formulations and manufacturers differed, the differences in tolerability, as well as platelet counts, might have been related to differences in pharmacokinetic properties between the 2 formulations. OBJECTIVES: The present series of investigations (1) determined the pharmacokinetic profile of anagrelide and its metabolites; (2) compared the pharmacokinetic profiles of the test and reference formulations of anagrelide; (3) investigated the in vitro release of anagrelide as a marker of intragastric anagrelide release of the test and reference formulations; and (4) compared the platelet-reducing effects of the test and reference formulations in patients with thrombocythemia in 2 longitudinal studies over 4 weeks. METHODS: A series of 4 in vivo studies and 1 in vitro study were conducted. In a pilot, prospective, singledose study in healthy volunteers, the pharmacokinetic properties (C(max), T(max), and AUC(0-infinity)) of a test formulation of anagrelide were assessed using high-performance liquid chromatography analysis of plasma samples. Based on the results from that study, a single-dose, randomized, double-blind, 2-period crossover study in healthy volunteers was conducted to determine bioequivalence of 2 formulations of anagrelide 2 mg/d (taken as 4 capsules). In vitro dissolution properties of the test or reference formulation containing 0.5 mg anagrelide as the active ingredient were studied in an assay mimicking gastrointestinal release. To test for effects on platelet counts of switching from the reference formulation (previous treatment on stable dose for 3 months) to the test formulation, two 4 week longitudinal trials were conducted: one in patients with ET (in Germany), and one in patients with thrombocythemia associated with chronic myeloproliferative disorders (CMPDs) (in Austria). RESULTS: The pilot pharmacokinetic study of the test formulation in 16 volunteers (10 women, 6 men; mean [SD] age, 20.5 [1.5] years; weight, 69.0 [10.0 kg) suggested that anagrelide was metabolized to 3-hydroxyanagrelide (AUC(0-infinity) 50% compared with anagrelide) and the inactive metabolite 2-amino-5,6-dichloro-,4 dihydroquinazolone. The subsequent bioequivalence study in 24 volunteers (14 women, 10 men; mean [SD] age, 23 [4] years; white, 100%; weight, 67.5 [10.2] kg) found that the test formulation was associated with a significantly lower C(max) (point estimation [PE], 66%; 90% CI, 58%-76%; P < 0.001) and AUC(0-infinity) (PE, 77%; 90% CI, 68%-86%; P = 0.001). T(max) values for anagrelide and 3 hydroxyanagrelide were 1 hour longer with the test formulation compared with the reference formulation. The total number of adverse events with the reference formulation was 46; the test formulation, 29 (P = 0.05). In vitro, anagrelide from the reference formulation was immediately released (89.1% at 5 minutes), whereas there was a delayed release (93.6% at 30 minutes) from the test formulation (P < 0.05). In the last 2 studies, 2 cohorts of white patients (cohort 1, 15 patients with ET; 10 women, 5 men; mean [SD] age, 49.0 [10.7] years [range, 31-66 years]; weight, 73.2 [12.6] kg; cohort 2, 19 patients with thrombocythemia associated with CMPD; 12 women, 7 men; age, 62.6 [12.4] years [range, 38-80 years]; weight, 66.1 [13.3] kg) who had received treatment for > or =3 months with the reference formulation were switched to the same dose of the test formulation and maintained on this dose for 4 weeks. Platelet counts did not change significantly from baseline over 4 weeks and stayed within a predefined margin of 150 x 10(3) cells/microL. CONCLUSIONS: The pharmacokinetic properties, adverse event rates, and in vitro dissolution profile differed between the test and reference anagrelide formulations in these healthy volunteers. In patients with ET or thrombocythemia associated with CMPD, platelet counts did not differ significantly from baseline at 4 weeks when subjects were switched from the reference to the test anagrelide formulation. PMID- 19302912 TI - Bioavailability of two oral-tablet and two oral-suspension formulations of naproxen sodium/paracetamol (acetaminophen): single-dose, randomized, open-label, two-period crossover comparisons in healthy Mexican adult subjects. AB - BACKGROUND: Naproxen sodium/paracetamol (acetaminophen) is a combination for the treatment of symptomatic pain and fever marketed both as a prescription and an over-the-counter product in Mexico. OBJECTIVE: The aim of these 2 studies was to compare the bioavailability and to determine the bioequivalence of 2 test formulations (an oral-tablet formulation containing the combination of naproxen sodium/paracetamol 275/300 mg and an oral-suspension formulation containing the combination of naproxen sodium/paracetamol 375/300 mg per 15 mL) with their corresponding listed reference-drug formulations in Mexico (a list issued by Mexican health authorities). METHODS: Two separate, single-dose, randomized, open label, 2-period crossover, postmarketing studies were conducted. For each study, a different set of eligible subjects was selected comprising healthy Mexican adults of either sex, and subjects were randomly assigned to receive 1 test formulation of the combination of naproxen sodium/paracetamol followed by the corresponding reference-drug formulation, or vice versa, with a 1-week washout period between doses. After a 12-hour overnight fast, subjects received a single dose of naproxen sodium/paracetamol 275/300-mg tablet or naproxen sodium/paracetamol 375/300 mg per 15 mL suspension, depending on the study. For the analysis of pharmacokinetic parameters, including C(max), AUC from time 0 (baseline) to 48 hours (AUC(0-48)), and AUC from baseline to infinity (AUC(0 infinity)), blood samples were drawn at baseline and at 0.16, 0.33, 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 4, 6, 8, 12, 24, and 48 hours after administration. The formulations were considered bioequivalent if the geometric mean ratios (test/reference) of the C(max) and AUC were within the predetermined range of 80% to 125%. Tolerability was determined by clinical assessment, monitoring vital signs, laboratory analysis results, and subject interviews regarding adverse events. RESULTS: A total of 26 subjects (15 men, 11 women; mean [SD] age, 29 [8] years [range, 20-50 years]; weight, 63.1 [9] kg [range, 51.4-84.4 kg]; height, 164 [9] cm [range, 149-179 cm]; and body mass index [BMI], 23.53 [2.18] kg/m(2) [range, 18.54-26.82 kg/m(2)]) were enrolled to receive the suspension-dosage formulation; 13 subjects received the suspension-test formulation first. A total of 26 subjects (13 men, 13 women; mean [SD] age, 29 [8] years [range, 18-43 years]; weight, 64.3 [7.7] kg [range, 50.6-80.7 kg]; height, 165 [9] cm [range, 151-181 cm]; and BMI, 23.64 [2.43] kg/m(2) [range, 18.02-26.42 kg/m(2)]) were enrolled to receive the tablet-dosage formulation; 13 subjects received the tablet-test formulation first. No significant period or sequence effects were detected based on analysis of variance. For the suspension-dosage formulation, the 90% CIs for naproxen C(max), AUC(0-48), and AUC(0-infinity) were 93.06% to 104.00%, 93.50% to 98.44%, and 92.14% to 98.99%, respectively, and were 90.09% to 105.90%, 88.58% to 99.34%, and 91.43% to 101.55%, respectively, for paracetamol. For the tablet-dosage formulation, the 90% CIs for naproxen C(max), AUC(0-48), and AUC(0-infinity) were 102.83% to 117.15%, 96.59% to 104.26%, and 96.01% to 102.90%, respectively, and were 94.04% to 121.09%, 95.48% to 105.64%, and 96.64% to 105.42%, respectively, for paracetamol. CONCLUSIONS: In these 2 small studies in healthy Mexican adult subjects, a single dose of naproxen sodium/paracetamol 275/300 mg of the test formulation of the tablet-dosage formulation or a single dose of naproxen sodium/paracetamol 375/300 mg per 15 mL of the test formulation of the suspension-dosage formulation was found to be bioequivalent to the corresponding reference formulations according to the regulatory definition of bioequivalence based on the rate and extent of absorption. All formulations were generally well tolerated. PMID- 19302913 TI - Bioequivalence of single 100-mg doses of two oral formulations of topiramate: an open-label, randomized-sequence, two-period crossover study in healthy adult male Mexican volunteers. AB - BACKGROUND: The proprietary form of topiramate is indicated in Mexico as an antiepileptic agent and in the prophylaxis of migraine headaches. However, before generic topiramate is placed on the market, pharmacokinetic studies investigating the bioequivalence of generic and branded formulations are needed. OBJECTIVE: The aim of this study was to compare the bioequivalence and tolerability of a generic (test) and a branded (reference) formulation of topiramate 100 mg in healthy Mexican volunteers. METHODS: This open-label, randomized-sequence, 2-period crossover study was conducted at Ipharma SA de CV, Monterrey, Mexico. Eligible subjects were healthy male Mexican volunteers aged 18 to 45 years. Participants were randomly assigned to receive 100 mg of the test or reference formulation, followed by a 3-week washout period and administration of the alternate formulation. Doses were administered after a 12-hour overnight fast. For analysis of pharmacokinetic properties, including C(max), AUC(0-t), and AUC(0-infinity), blood samples were obtained over a 144-hour period after dosing. The formulations were to be considered bioequivalent if calculations of a 90% CI for the ratio of the means of the measures for the test and reference formulations fell within bioequivalence limits, 80% to 125%, for logarithmic (log) transformation of C(max) and AUC, and if two 1-sided t tests showed P < 0.05. Tolerability was assessed using vital sign measurement (blood pressure, body temperature, heart rate, and respiratory rate), laboratory analysis (hematology, blood biochemistry, hepatic function, and urinalysis), and subject interview. RESULTS: Twenty-eight men (mean age, 22.21 years [range, 18-28 years]; mean weight, 75.04 kg [range, 62 96 kg]; mean height, 177 cm [range, 163-192 cm]) were enrolled in this study, and 28 (14 each randomized to receive the test or reference formulation first) completed it. No period or sequence effects were observed. The 90% CIs for the log-transformed C(max), AUC(0-t) and AUC(0-infinity) were 94.70 to 112.05, 98.88 to 105.16, and 98.80 to 105.28, respectively (all, P < 0.05). No adverse events were reported by the volunteers or found on clinical laboratory testing during the study. CONCLUSIONS: This study did not find any statistically significant differences in C(max) or AUC values between the test and reference formulations of oral topiramate 100 mg in this population of healthy adult male Mexican volunteers. On that basis, and according to both the rate and extent of absorption, the test and reference formulations met the regulatory criteria for bioequivalence. Both formulations were well tolerated. PMID- 19302914 TI - Pharmaceutical use and outcomes in children. PMID- 19302915 TI - A framework for planning and critiquing medication compliance and persistence research using prospective study designs. AB - BACKGROUND: Medication compliance and persistence are important determinants of clinical outcomes. With the application of evidence-based therapy, it is increasingly important to ensure that studies that use compliance or persistence as a primary or secondary outcome are designed suitably and employ appropriate analyses to support the inferences made. OBJECTIVE: The aim of this work was to describe the designs of medication compliance/persistence studies and provide guidance on appropriate analyses, with the ultimate goal of helping health providers and payers of health care understand the impact of compliance and persistence on health outcomes. METHODS: MEDLINE, CINAHL, EMBASE, and all EBM Reviews databases were searched to locate key research articles about prospective medication compliance and persistence studies. Articles published between 1978 and 2008 were included in the search. Inclusion criteria included a focus on medication compliance and persistence, and prospective research designs. Articles that largely focused on retrospective study designs or were based on opinion rather than evidence were excluded. RESULTS: A systematic framework was developed that comprised a prospective checklist and a quantitative tool to assess the quality of studies. The key elements of the checklist included the following: title and abstract, introduction or background, objectives, methods and study design, statistical analysis and results, discussion, conclusions, and disclosure of conflicts of interest. For each element, examples are provided to help readers make an informed decision about the design, value, and quality of a particular prospective study. CONCLUSIONS: The checklist and quantitative tool can be used to provide objective validation of the rigor of prospective research designs. It is anticipated that future research will follow a uniform approach to presentation and evaluation of data, thereby facilitating a clear understanding of the impact of compliance and persistence on health outcomes. PMID- 19302916 TI - Estimates of pediatric medication use in the United States: current abilities and limitations. AB - BACKGROUND: Resources are available for measuring adult medication use, but similar resources have not been fully developed for measuring pediatric use. Policy decisions require an understanding of the population affected, the number of children, their ages, sex, geographic distribution, race and ethnicity, and insurance status, as well as trends over time. OBJECTIVE: In this article, databases providing information about prescription drugs used in the United States are reviewed with respect to pediatric populations. METHODS: A series of searches were conducted in MEDLINE using these terms: frequency, prevalence, drug utilization, children, pediatric, drug usage, medications, and prescriptions. Authors of selected articles were interviewed to identify salient issues in the measurement of pediatric medication use. Preliminary analysis of several databases followed within the context of government implementation of the Best Pharmaceuticals for Children Act. This was followed by further MEDLINE searches and synthesis of the literature. RESULTS: Databases with information about pediatric population medication use included 7 with outpatient data and 4 with inpatient data. Outpatient data were available from government and private sources, but inpatient data were available from private sources only. Three of the government and 1 of the private databases with outpatient data had sample sizes of several thousand, too small to allow analysis of frequency trends in pediatric populations or subpopulations, in which many drugs are used by fewer than 0.01% of patients. CONCLUSIONS: Sample size needs are greater when measuring pediatric medication use because the overall level of use is lower among children than adults. Databases resulting from hospital quality efforts, conglomeration of pharmacy benefit records, and standardization of state Medicaid records offer opportunities to describe prescription medication use in samples of several hundred thousand to several million children but will require dedicated resources. PMID- 19302917 TI - Silencing of genes required for glycosylphosphatidylinositol anchor biosynthesis in Burkitt lymphoma. AB - OBJECTIVE: To investigate the mechanism of glycosylphosphatidylinositol (GPI) anchor deficiency in Burkitt lymphoma cell lines. METHODS: We identified a large GPI anchor protein deficient population in three different Burkitt lymphoma cell lines through proaerolysin treatment of the cells and flow cytometry analysis using a proaerolysin variant (FLAER). The mechanism of GPI anchor protein deficiency was studied by DNA gene sequencing, a cell-free assay to investigate the GPI anchor biosynthetic pathway, microarray analysis, and quantitative real time polymerase chain reaction. RESULTS: Burkitt lymphoma cell lines harbor large populations of FLAER(neg) cells, which are resistant to proaerolysin. In all three cell lines, silencing of a gene involved in an early step in GPI-anchor biosynthesis was responsible for the lack of GPI-anchored proteins on the cell surface. Quantitative polymerase chain reaction and microarray analysis demonstrate that the level of mRNA for PIGL and PIGY is lower in the FLAER(neg) Ramos cells and that mRNA levels of PIGY are reduced in the Akata and Daudi cells. Hypermethylation of these genes was associated with the low levels of mRNA and treatment of the cells with 5-aza-2' deoxycytidine restored cell surface GPI anchored proteins to the FLAER(neg) cells. CONCLUSION: GPI-anchored protein deficiency in Burkitt lymphoma cells is not due to a genetic mutation (e.g., PIGA); rather, the lack of GPI-anchored proteins results from transcriptional silencing of PIGL and PIGY. PMID- 19302918 TI - Stem cells of GATA1-related leukemia undergo pernicious changes after 5 fluorouracil treatment. AB - OBJECTIVE: Transcription factor GATA1 plays a critical role in erythropoiesis through the integrated regulation of cell proliferation, differentiation, and apoptosis. In Gata1.05 gene knockdown mice, Gata1 expression deteriorates to 5% of wild-type allelic expression, a level insufficient for supporting normal erythropoiesis and one that leads to accumulation of erythroid progenitors that are readily transformed into erythroblastic leukemia. Serial engraftment of leukemic cells into primary or subsequent nude mice reconstituted complete leukemic phenotype in recipient. To delineate characteristics of leukemic stem cells (LSCs), we analyzed LSCs of Gata1.05 leukemia, which have a potential to reestablish leukemia in mice. MATERIALS AND METHODS: Leukemic cells isolated from the first recipient mice of Gata1.05 leukemia cells were divided into two fractions using Hoechst dye. Fractionated cells were transplanted into second recipient, or analyzed gene expression profiles and cell-cycle status. Consequences of 5-fluorouracil (5-FU) treatment on leukemic cells in vivo were studied. RESULTS: LSCs were enriched in the Hoechst dye-excluded side population (SP), and leukemic cells in the SP population (LSP cells) were morphologically and immunophenotypically indistinguishable from other leukemic cells. However, expression of hematopoietic stem cell (HSC)-related genes was upregulated in the LSP cells. In cell-cycle analyses, LSP cells were quiescent like HSCs, but reentry into the cell cycle was stimulated by 5-FU treatment. Nonetheless, 5-FU treatment established a point of newly adjusted equilibrium in the LSP cells and the cells never recovered to their previous quiescent state. CONCLUSION: Based on this observation, distinct self-renewal regulatory mechanisms in LSCs may be considered as one of the causes of worsening of the features of leukemia after injury and relapse. PMID- 19302919 TI - Combined inhibition of integrin linked kinase and FMS-like tyrosine kinase 3 is cytotoxic to acute myeloid leukemia progenitor cells. AB - OBJECTIVE: Dysregulation of signaling pathways leading to enhanced cell proliferation and resistance to apoptosis is frequent in acute myeloid leukemia (AML). The effectiveness of inhibiting two such pathways, the phosphatidylinosityl-3-kinase pathway via the intermediate integrin-linked kinase (ILK), and FMS-like tyrosine kinase-3 (FLT-3) signaling pathway in killing AML cells was studied. MATERIALS AND METHODS: AML colony-forming cell (CFC) assays were used to determine the effects of a small molecule inhibitor of both ILK and FLT-3 (QLT0267) on poor prognosis primary AML sample viability. Kinase assays and Western blots were used to analyze effects of the compound on target molecules. RESULTS: In 31/36 AML blast samples p-Akt was detected indicating phosphatidylinosityl-3-kinase activation. ILK was ubiquitously and FLT-3 abundantly expressed. Downregulation of ILK in the AML cell line TF-1 using small interfering RNA caused >or= 50% CFC death, suggesting ILK inhibition might also be toxic to primary AML cells. In vitro kinase assays on three AML samples showed inhibition of both ILK and FLT-3 by QLT0267. Treatment of AML patient blast cells (n=27) with QLT0267, caused a dose- and time-dependent downregulation of p-Akt and kill of AML-CFC with AML samples containing FLT-3 mutations being more sensitive to QLT0267 than those without. AML samples were more sensitive to QLT0267 killing than normal bone marrow (IC(50)=3 microM, vs 10 microM for AML CFC and normal CFC, respectively, n=5). CONCLUSION: Combined inhibition of ILK and FLT-3 with a small molecule kinase inhibitor can achieve selective targeting of AML rather than normal hematopoietic progenitors. PMID- 19302920 TI - Effects of imatinib mesylate in osteoblastogenesis. AB - Imatinib mesylate (IM), a tyrosine kinase inhibitor currently used in chronic myeloid leukemia (CML), may also affect the growth of other cellular systems besides CML cells. Because it has been reported that IM may affect bone tissue remodeling, we evaluated the effects of IM on osteoblastic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs). After 21 days of culture, hBM-MSCs treated with IM (1 microM) alone or osteogenic medium (OM) + IM showed changes in morphology with evidence of extracellular mineralization and increased mRNA expression of osteogenic markers, such as RUNX2, osteocalcin (OCN), and bone morphogenetic protein (BMP-2). We also observed that levels of OCN and the osteoprotegerin (OPG)/receptor activator of nuclear factor-kappa B ligand (RANKL) ratio (OPG/RANKL ratio) were increased in the surnatant of the 21-day culture with IM or OM + IM compared to controls (p<0.005). In addition, we found that in 46 serum samples collected from CML patients treated with IM for 3 to 24 months, the OPG/RANKL ratio increased after 3 and 6 months (p<0.004) returning back to the basal level after 24 months of IM treatment. In these patients, OCN levels were low at diagnosis but they increased throughout the IM treatment, approaching normal levels at 24 months of IM therapy. In summary, our data show that IM increases mRNA expression of osteogenic markers in hBM-MSCs and increases the OPG/RANKL ratio and the OCN levels both in surnatant of hBM-MSCs cultured with IM and in serum of patients treated with IM, thus indicating that IM potentially favors osteoblastogenesis. PMID- 19302921 TI - Establishment of an animal model for Waldenstrom's macroglobulinemia. AB - OBJECTIVE: Waldenstrom's macroglobulinemia (WM) is a low-grade lymphoplasmacytoid lymphoma characterized by production of monoclonal immunoglobulin M (IgM). The present study was undertaken with the aim of developing a novel nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse model of WM. MATERIALS AND METHODS: Pairs of bone particles derived from adult humans were successfully implanted intramuscularly in NOD/SCID mice. Each mouse was implanted with a bone fragment taken from a neoplastic disease-free individual in the one hind limb and with a different biopsy taken from a WM patient in the other. RESULTS: All mice implanted with the bone marrow core biopsies had increased levels of serum IgM 1 month following the implantation onward. Histopathologic and immunohistochemical analysis showed that in approximately half of the mice WM cells metastasized from the WM bone implant to the distantly implanted non-WM bone. Serum IgM value records of all mice correlated with histopathological observations and immunohistochemical analysis for neoplastic cell density and metastatic growth. CONCLUSION: Results obtained in the present study suggest that IgM-producing WM cells not only retained viability in the bone marrow of the WM bone biopsy, but also metastasized to the normal bone marrow of the distant bone implant. The mouse model reported here improves on existing models of WM by recapitulating the adult human bone marrow microenvironment of abnormal WM neoplastic cells. PMID- 19302922 TI - Compound heterozygous c-Mpl mutations in a child with congenital amegakaryocytic thrombocytopenia: functional characterization and a review of the literature. AB - OBJECTIVE: To genetically and functionally characterize mutations of c-Mpl that lead to thrombocytopenia in a child with congenital amegakaryocytic thrombocytopenia. MATERIALS AND METHODS: We identified two c-Mpl mutations in a child with clinical features of congenital amegakaryocytic thrombocytopenia, one a previously described mutation in the extracellular domain (R102P) and the other a novel mutation leading to truncation of the receptor after the box 1 homology domain (541Stop). Cell line models were created to examine the ability of the mutant receptors to signal in response to thrombopoietin and thrombopoietin-like agonists. RESULTS: Data from cell-line models indicate that c-Mpl R102P does not support significant signaling in response to thrombopoietin due to impaired trafficking of the mutant receptor to the cell surface. Alternative thrombopoietic agents do not circumvent this block to signaling, likely due to the inaccessibility of the receptor. In addition, previous data indicate that c Mpl 541Stop does not support intracellular signaling due to the loss of critical intracellular domains. CONCLUSIONS: This case demonstrates two different mechanisms by which c-Mpl mutations can impair thrombopoietin signaling, and suggests that mutations in the extracellular domain will not be rescued by c-Mpl agonists if they interfere with normal receptor expression. PMID- 19302923 TI - Upregulation of plasma CCL8 in mouse model of graft-vs-host disease. AB - OBJECTIVE: Using a proteomic approach, we recently identified plasma CCL8 as a potential biomarker for diagnosis of graft-vs-host-disease (GVHD) in mice as well as humans. Because mass spectrometric analysis is only semi-quantitative, a quantitative method of measuring plasma CCL8 levels in mice is needed. MATERIALS AND METHODS: We established an enzyme-linked immunosorbent assay for the quantitative measurement of CCL8 concentrations in mouse plasma. RESULTS: Our newly established enzyme-linked immunosorbent assay revealed that the plasma CCL8 concentrations (mean +/- standard error; n=12) were 1287+/-55.7 ng/mL and 1604+/ 110.8 ng/mL on days 7 and 14 after allogeneic bone marrow transplantation (BMT), respectively, while the plasma concentrations was 316.6+/-16.3 ng/mL on day 7 after syngeneic BMT. A Western blotting analysis also showed a difference in the plasma CCL8 levels between the allogeneic and syngeneic BMT groups, as did clinical GVHD scores. Neither lipopolysaccharide nor poly(I:C) elevated the plasma CCL8 concentrations, although a dramatic increase in interleukin-6 was detected after both treatments. CONCLUSION: An elevated plasma CCL8 concentration may be a promising plasma marker for GVHD in mouse models. PMID- 19302924 TI - A comparison of the fungicidal activity of amphotericin B and posaconazole against Zygomycetes in vitro. AB - Zygomycoses is a rapidly progressive infection associated with high mortality. Although amphotericin B (AMB) has been the only treatment option for years, recent studies have demonstrated that posaconazole (PCZ) has good activity against Zygomycetes. Because rapid onset of antifungal activity is crucial in the management and never studied before, we compared the time for maximum fungicidal activity of AMB versus PCZ using time-kill curves. The MIC of AMB and PCZ against clinical isolates of Mucor spp. and Rhizopus spp. was determined by the Clinical and Laboratory Standards Institute M38-A2 method, and the fungicidal activity was examined by time-kill studies. AMB was rapidly fungicidal, with 95% killing noted as early as 6 h and 99.9% killing at 24 h; PCZ showed <70% killing at 6 h and 99.9% killing at 48 h. In vivo animal studies using AMB in the early phase, followed by a switch to high-dose PCZ later, could provide data that may have clinical implications because there are only a few drugs currently available for the management of zygomycoses. PMID- 19302925 TI - A rare case of silicone mammary implant infection by Streptomyces spp. in a patient with breast reconstruction after mastectomy: taxonomic characterization using molecular techniques. AB - A Streptomyces sp. isolated from a patient who had had breast reconstruction after a mastectomy was identified at the species level by comparative sequence analysis of 16S ribosomal DNA (rDNA) and the hypervariable alpha-region of the 16S rDNA. PMID- 19302926 TI - Investigation of carbapenem-resistant Acinetobacter baumannii isolates in a district hospital in Taiwan. AB - A total of 34 Acinetobacter baumannii isolates from a district hospital in Taiwan were identified with carbapenem-hydrolyzing oxacillinase OXA-66/OXA-51-like. In addition, 26 of 28 carbapenem-resistant isolates harbored plasmid-encoded bla(OXA 23)-like genes. Twenty of 28 carbapenem-resistant isolates mapped to the major genotype cluster A of carbapenemase producer by pulsed-field gel electrophoresis. PMID- 19302927 TI - In vitro potency of doripenem tested against an international collection of rarely isolated bacterial pathogens. AB - Doripenem, a new 1beta-methyl parenteral carbapenem, has very broad-spectrum activity against Gram-positive and Gram-negative aerobic bacteria. As noted here, the spectrum and potency extended to many rarely isolated species sampled by the Doripenem Global Surveillance Program. Among the species or species groups with 0.25 microg/mL for all tested species except Lactococcus garvieae, Listeria monocytogenes, and Micrococcus spp. In conclusion, doripenem exhibited a very wide spectrum but variable potencies against uncommonly cultured aerobic bacterial pathogens isolated in 2003 to 2007. These results confirm the potential use of this new carbapenem for broad-spectrum empiric or directed antimicrobial therapy. PMID- 19302928 TI - Antimicrobial activity of doripenem tested against prevalent Gram-positive pathogens: results from a global surveillance study (2003-2007). AB - Doripenem is a broad-spectrum parenteral carbapenem recently approved in the United States for treatment of complicated urinary tract and intra-abdominal infections. Although pronounced doripenem antimicrobial activity against various Gram-negative pathogens, including anaerobes, has been confirmed, limited information has been forthcoming on the activity of this agent against leading Gram-positive species. We evaluated the activity of doripenem using reference broth microdilution procedures against a large collection of staphylococci, enterococci, and streptococci collected as part of a global (North America, 43.0%; Latin America, 11.7%; Europe, 31.3%; and Asia-Pacific, 14.0%) Doripenem Surveillance Program for the years 2003 to 2007. Doripenem was confirmed to be highly active against oxacillin-susceptible Staphylococcus aureus (22 389 isolates) and coagulase-negative staphylococci (2444 isolates; MIC(90) values, 90%) of macrofollicles with a surface area that was up to 5 times larger than the surface area of normal follicles, as calculated with an image analysis system. Capsular invasion was detected in 2 separate foci. The tumor was classified as a minimally invasive follicular carcinoma, macrofollicular variant. This case is detailed to highlight the potential pitfall that may arise from an incomplete histological analysis of a macrofollicular lesion, with particular attention paid to the differential diagnoses. PMID- 19302959 TI - Grossing biopsies: an introduction to general principles and techniques. AB - Representation of grossing biopsies methodology is minimal in the surgical pathology literature. The present review is an attempt to provide an update on general principles and common techniques for grossing small biopsies. Biopsy triage is the first step of grossing that facilitates optimal handling of the specimen. It includes determination of the specimen's category, priority for processing, and adequacy of the specimen's preservation. New requirements of modern biopsy processing, such as the numerous modern ancillary studies and increased expectations for rapid turnaround time, make this often underestimated preanalytical phase more significant. This article discusses general requirements for biopsy grossing techniques that include completeness of submission, selecting for microscopy the most diagnostically valuable parts of the biopsy specimen, and specific instructions for further histology processing. PMID- 19302960 TI - Osteoclastlike giant cell tumor of the salivary gland. AB - Giant cell tumor of the salivary gland is extremely rare, with only 15 cases published in the English literature. The tumor characteristically contains a mixture of multinucleated giant cells, resembling osteoclasts of bone, and neoplastic mononuclear cells. In about half of the reported cases, there is an associated carcinomatous component. We are reporting an additional case of giant cell tumor of the parotid gland that was initially misinterpreted as an extraosseous osteosarcoma in the biopsy specimen. The histologic and immunohistochemical findings as well as a review of the literature with discussion of the histogenesis of this unusual neoplasm are presented. PMID- 19302961 TI - Clinical features and differential diagnoses of solitary extramedullary plasmacytoma of the thyroid: a case report. AB - A 40-year-old woman presented with a rapidly enlarging palpable thyroid mass. The patient underwent a total thyroidectomy. The tumor fulfilled the criteria of primary solitary extramedullary plasmacytoma (SEP), including cellular expression of the CD138 and lambda light chain antibodies. Solitary extramedullary plasmacytoma of the thyroid occurs most commonly in patients with Hashimoto thyroiditis and must be distinguished from involvement of thyroid in multiple myeloma, inflammatory pseudotumor plasma cell variant, mucosa-associated lymphoid tissue lymphoma, and medullary carcinoma. The distinction is determined on the basis of histologic findings, immunohistochemical analysis, and other laboratory tests. Currently, no standard treatment exists for this entity. In this report, we discuss the differential diagnosis of SEP of the thyroid and the clinical features observed in this case. PMID- 19302962 TI - Paraneoplastic sensory neuropathy associated with small cell carcinoma of the gallbladder. AB - A small number of patients with carcinoma of the gallbladder can present with a variety of paraneoplastic syndromes, including the Cushing syndrome, hypercalcemia, acanthosis nigricans, bullous pemphigoid, dermatomyositis, and the Leser-Trelat sign. We report on what appears to be the first case of a patient, a 48-year-old woman, with anti-Hu paraneoplastic sensory neuropathy and small cell carcinoma of the gallbladder. The patient's neurologic symptoms preceded the diagnosis of small cell carcinoma by 11 months. These symptoms improved after surgical removal of the tumor and chemotherapy. The small cell carcinoma was relatively small and was not associated with gallstones. In spite of the small size of the tumor, it metastasized to a celiac lymph node and probably to the liver. Anti-Hu paraneoplastic sensory neuropathy should be added to the list of paraneoplastic syndromes associated with small cell carcinoma of the gallbladder. PMID- 19302963 TI - Sclerosing extramedullary hematopoietic tumor: emphasis on diagnosis by renal biopsy. AB - Sclerosing extramedullary hematopoietic tumor (SEMHT) is a rare lesion that typically arises in patients with chronic myeloproliferative disorders. Morphologically, it exhibits atypical megakaryocytes, granulocytic precursors, and erythroid precursors set in a background of dense collagen sclerosis. Sclerosing extramedullary hematopoietic tumor may be easily mistaken for other neoplasms, such as sarcoma, carcinoma, or Hodgkin lymphoma, particularly if pertinent clinical history is not provided. Misdiagnosis may occur because of the difficulty in recognizing the megakaryocytic lineage of the atypical cells and because of the paucity of other hematopoietic elements. We report a case of a 72 year-old man with proteinuria and renal insufficiency who underwent renal biopsy to determine the etiology of the proteinuria. The kidney biopsy demonstrated an unusual tumor in which the initial morphological impression that of sclerosing liposarcoma. However, upon learning of the patient's previous history of chronic idiopathic myelofibrosis and with the aid of immunohistochemistry, the correct diagnosis of sclerosing extramedullary hematopoietic tumor was made. Sclerosing extramedullary hematopoietic tumor should be considered in the differential diagnosis when percutaneous renal biopsy or other intra-abdominal biopsy reveals a sclerotic lesion with interspersed large atypical cells, especially in a patient with a history of chronic myeloproliferative disorder. PMID- 19302964 TI - Pseudomembranous disease (ligneous inflammation) of the female genital tract, peritoneum, gingiva, and paranasal sinuses associated with plasminogen deficiency. AB - Pseudomembranous disease (or ligneous inflammation) is a rare condition characterized by accumulations of fibrin-rich eosinophilic material. Recent investigations have linked the etiology of this condition to plasminogen deficiency (hypoplasminogenemia). Although much of the literature concerning this disease has focused upon the often clinically striking ocular manifestations, it is important to note that pathologic changes may develop in a variety of anatomic locations, including the oral cavity, upper and lower respiratory tract, female genital tract, kidneys, and gastrointestinal tract. Here, we report an unusual case of a 33-year-old woman who initially presented with gingival inflammation. In subsequent years, she developed additional signs and symptoms related to sinonasal and genital tract involvement. Despite numerous clinical evaluations, biopsies, and laboratory tests, the patient's diagnosis remained elusive for 7 years. Ultimately, it was the distinctive appearance of the gingiva that led to a diagnosis of plasminogen deficiency. Unfortunately, the complicated clinical course and elapsed time between initial presentation and diagnosis illustrated by the present case are not uncommon among patients with this condition. Greater familiarity with the clinical and histopathologic features of this condition among pathologists and treating clinicians is essential for timely diagnosis and management. PMID- 19302965 TI - Molecular biology: an early detector of oral cancers. AB - Oral cancers have been one of the leading causes of deaths particularly in the developing countries. Prime reason for this high mortality and morbidity is attributed to the delay in diagnosis and prompt treatment. Relentless research in the field of oncology has led to advent of novel procedures for the early detection of oral cancers. Molecular biology is highly promising in this regard. It is a procedure that detects alterations at a molecular level much before they are seen under a microscope and much before clinical changes occur. Molecular studies serve as basis by which we will eventually be able not only to augment clinical assessment and classification of oral lesions but also predict malignant potential of oral lesions, thus reducing incidence and increasing the scope for early diagnosis and treatment of oral cancers. However, making such sophisticated tools available for the common man in developing countries is one of the most important challenges faced today. PMID- 19302966 TI - Journal of Pediatric Urology. Editorial. PMID- 19302967 TI - Pelvi-ureteric junction obstruction with crossing renal vessels: a case report of failed laparoscopic vascular hitch. AB - INTRODUCTION: Dismembered pyeloplasty has been the traditional technique in the management of pelvi-ureteric junction obstruction (PUJO) secondary to crossing renal vessels in children. Laparoscopic transposition of lower pole vessels for PUJO has been described in children as well as adults. CASE REPORT: We report a child with PUJO secondary to lower pole renal vessels who underwent laparoscopic transposition of vessels. The child had persistent PUJO, which was later treated with laparoscopic dismembered pyeloplasty. CONCLUSION: Laparoscopic transposition of renal vessels is a simple technique, and requires less operating time. It involves no anastomosis or insertion of temporary DJ stent. However, as of now there are no imaging techniques or intraoperative procedures available to confirm that the crossing renal vessels are the sole etiology for PUJO. This option should therefore be used with caution. PMID- 19302968 TI - Number matters: control of mammalian mitochondrial DNA copy number. AB - Regulation of mitochondrial biogenesis is essential for proper cellular functioning. Mitochondrial DNA (mtDNA) depletion and the resulting mitochondrial malfunction have been implicated in cancer, neurodegeneration, diabetes, aging, and many other human diseases. Although it is known that the dynamics of the mammalian mitochondrial genome are not linked with that of the nuclear genome, very little is known about the mechanism of mtDNA propagation. Nevertheless, our understanding of the mode of mtDNA replication has advanced in recent years, though not without some controversies. This review summarizes our current knowledge of mtDNA copy number control in mammalian cells, while focusing on both mtDNA replication and turnover. Although mtDNA copy number is seemingly in excess, we reason that mtDNA copy number control is an important aspect of mitochondrial genetics and biogenesis and is essential for normal cellular function. PMID- 19302969 TI - Differentiation potential of bone marrow mesenchymal stem cells in duck. AB - The bone marrow mesenchymal stem cells (MSCs) are multipotent stem cells which can differentiate into mesenchymal cells in vitro. In this study, MSCs in duck were isolated from bone marrow by density gradient centrifuge separation, purified and expanded in the medium. The primary MSCs were expanded for passages. The different-passage MSCs were induced to differentiate into osteoblasts and neuron-like cells. Karyotype analysis indicated that MSCs kept diploid condition and the hereditary feature was stable. The different-passage MSCs expressed CD44, ICAM- and SSEA-4, but not CD34, CD45 and SSEA-when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 2, 6 and 8 (P > 0.05), but a significant difference existed among those of passages 2, 6, 8 and 11 (P < 0.05). After the osteogenic inducement was added, the induced different-passage MSCs expressed high-level alkaline phosphatase (ALP), and are positive for tetracycline staining, Alizarin Red staining and Von Kossa staining. After the neural inducement was added, about 70% cells exhibited typical neuron-like phenotype, the induced different-passage MSCs expressed Nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) when detected by immunofluorescence staining. There was no significant difference among the positive rates of passages 3, 4 and 6 (P>0.05), but a significant difference existed among those of passages 3, 4, 6 and 8 (P<0.05). These results suggest that MSCs in duck were capable of differentiating into osteoblasts and neuron-like cells in vitro. PMID- 19302970 TI - Cotton GhPOX1 encoding plant class III peroxidase may be responsible for the high level of reactive oxygen species production that is related to cotton fiber elongation. AB - The accumulation of reactive oxygen species (ROS) is involved in plant cell development. In plant, class III peroxidases are heme-containing enzymes encoded by a large multi-gene family participated in the release or consumption of ROS. The specific function of each member of the family is still elusive. Here, we showed that ROS was significantly generated during cotton fiber initiation and elongation, whereas, application of NADPH oxidase inhibitor diphenyleneiodonium (DPI) and peroxidase inhibitor salicylhydroxamic acid (SHAM) to the wild-type cotton ovule culture significantly suppressed fiber growth, respectively. Their inhibitory effects were caused by the reduction of superoxide radical (O(2)(-)). Ten GhPOX genes (cDNAs) encoding cotton class III peroxidases were isolated, among them eight GhPOX genes were reported for the first time. Microarray analyses indicated that GhPOX1 was the mostly predominantly expressed in fast elongating cotton fiber cells. Real-time quantitative PCR analysis revealed the transcript level of GhPOX1 was over 400-fold higher in growing fiber cells than in ovules, flowers, roots, stems and leaves. To reveal the role of GhPOX1 in plant development, its Arabidopsis orthologue atpox13 mutant was demonstrated to be defective in branch root development. Taken together, the data suggest that GhPOX1 plays an important role during fiber cell elongation possibly by mediating production of reactive oxygen species. PMID- 19302971 TI - Revaluation of deuterostome phylogeny and evolutionary relationships among chordate subphyla using mitogenome data. AB - The traditional knowledge in textbooks indicated that cephalochordates were the closest relatives to vertebrates among all extant organisms. However, this opinion was challenged by several recent phylogenetic studies using hundreds of nuclear genes. The researchers suggested that urochordates, but not cephalochordates, should be the closest living relatives to vertebrates. In the present study, by using data generated from hundreds of mtDNA sequences, we revalue the deuterostome phylogeny in terms of whole mitochondrial genomes (mitogenomes). Our results firmly demonstrate that each of extant deuterostome phyla and chordate subphyla is monophyletic. But the results present several alternative phylogenetic trees depending on different sequence datasets used in the analysis. Although no clear phylogenetic relationships are obtained, those trees indicate that the ancient common ancestor diversified rapidly soon after their appearance in the early Cambrian and generated all major deuterostome lineages during a short historical period, which is consistent with "Cambrian explosion" revealed by paleontologists. It was the 520-million-year's evolution that obscured the phylogenetic relationships of extant deuterostomes. Thus, we conclude that an integrative analysis approach rather than simply using more DNA sequences should be employed to address the distant evolutionary relationship. PMID- 19302972 TI - Molecular evolution and functional divergence of HAK potassium transporter gene family in rice (Oryza sativa L.). AB - The high-affinity K(+) (HAK) transporter gene family is the largest family in plant that functions as potassium transporter and is important for various aspects of plant life. In the present study, we identified 27 members of this family in rice genome. The phylogenetic tree divided the land plant HAK transporter proteins into 6 distinct groups. Although the main characteristic of this family was established before the origin of seed plants, they also showed some differences between the members of non-seed and seed plants. The HAK genes in rice were found to have expanded in lineage-specific manner after the split of monocots and dicots, and both segmental duplication events and tandem duplication events contributed to the expansion of this family. Functional divergence analysis for this family provided statistical evidence for shifted evolutionary rate after gene duplication. Further analysis indicated that both point mutant with positive selection and gene conversion events contributed to the evolution of this family in rice. PMID- 19302973 TI - Root and shoot traits responses to phosphorus deficiency and QTL analysis at seedling stage using introgression lines of rice. AB - Phosphorous (P) deficiency is a major restraint factor for crop production and plants have developed several mechanisms to adapt to low P stress. In this study, a set of 271 introgression lines (ILs) were used to characterize the responses of seedlings to low P availability and to identify QTLs for root traits, biomass, and plant height under P-deficiency and P-sufficiency conditions. Plant height, total dry weight, shoot dry weight, and root number were inhibited under P deficiency, whereas maximum root length (MRL) and root-shoot ratio (RS) were induced by P-deficiency stress. Relative MRL (RMRL, the ratio of MRL under P deficiency to MRL under P-sufficiency condition) and relative RS (RRS) were used to evaluate P-deficiency tolerance at the seedling stage. A total of 24 additive QTLs and 29 pairs of epistatic QTLs were detected, but only qRN4 was detected in both conditions. This suggested that different mechanisms may exist in both P supply levels. QTLs for adaptive traits (RMRL, RRS, RRV, and RRDW) and qRN4 consistently expressed to increase trait stability may contribute to P-deficiency tolerance. Twelve intervals were cluster regions of QTLs for P-deficiency tolerance, and one QTL (qRRS8) showed pleiotropic effects on P-deficiency tolerance and drought tolerance. These interesting QTLs can be used in marker assisted breeding through the target ILs. PMID- 19302974 TI - Total RNA isolation from recalcitrant yeast cells. AB - Conventional methods of RNA isolation are not suitable for yeast cells from stationary phase and fermentation broth. Methods specially reported for such cells are cumbersome and do not lend themselves for use with large number of samples. Here we report a facile method of RNA isolation from such recalcitrant yeast cells. The entire procedure is performed in microcentrifuge tubes and, thus, is ideal for faster processing of multiple samples. The method consistently gives high quality and quantity of RNA, which was found to be suitable for downstream applications such as quantitative real-time polymerase chain reaction. Besides Saccharomyces cerevisiae, the method was found to work equally well with other yeast species; thus, it is likely to have wider applicability. PMID- 19302975 TI - CXCR4/SDF-1 pathway is crucial for TLR9 agonist enhanced metastasis of human lung cancer cell. AB - Accumulating data suggested that CXCR4/SDF-1 pathway may play an important role in the metastasis of tumor. We previously demonstrated that CpG ODN could enhance the metastasis of human lung cancer cell via TLR9. Here we further evaluated the possible role of CXCR4/SDF-1 pathway in the enhanced metastasis of human lung cancer 95D cells induced by CpG ODN. Our data showed down-regulation of CXCR4 expression using siRNA against CXCR4 could significantly reduce the enhanced metastasis of 95D cells induced by CpG ODN both in vitro and in vivo. These results suggested that TLR9 agonist might promote the metastasis of human lung cancer cells via CXCR4/SDF-1 pathway. PMID- 19302976 TI - Regulation of tyrosine phosphatases in the adventitia during vascular remodelling. AB - Protein tyrosine phosphatases (PTPs) are regulators of growth factor signalling in vascular remodelling. The aim of this study was to evaluate PTP expression in the context of PDGF-signalling in the adventitia after angioplasty. Utilising a rat carotid artery model, the adventitial layers of injured and non-injured vessels were laser microdissected. The mRNA expression of the PDGF beta-receptor, the ligands PDGF-A/B/C/D and the receptor-antagonising PTPs (DEP-1, TC-PTP, SHP 2, PTP1B) were determined and correlated to vascular morphometrics, proliferation markers and PDGF beta-receptor phosphorylation. The levels of the PDGF beta receptor, PDGF-C and PDGF-D were upregulated concurrently with the antagonising PTPs DEP-1 and TC-PTP at day 8, and normalised at day 14 after vessel injury. Although the proliferation parameters were time-dependently altered in the adventitial layer, the phosphorylation of the PDGF beta-receptor remained unchanged. The expression dynamics of specific PTPs indicate a regulatory role of PDGF-signalling also in the adventitia during vascular remodelling. PMID- 19302977 TI - MicroRNA-21 directly targets MARCKS and promotes apoptosis resistance and invasion in prostate cancer cells. AB - Prostate cancer is one of the most common malignant cancers in men. Recent studies have shown that microRNA-21 (miR-21) is overexpressed in various types of cancers including prostate cancer. Studies on glioma, colon cancer cells, hepatocellular cancer cells and breast cancer cells have indicated that miR-21 is involved in tumor growth, invasion and metastasis. However, the roles of miR-21 in prostate cancer are poorly understood. In this study, the effects of miR-21 on prostate cancer cell proliferation, apoptosis, and invasion were examined. In addition, the targets of miR-21 were identified by a reported RISC coimmunoprecipitation-based biochemical method. Inactivation of miR-21 by antisense oligonucleotides in androgen-independent prostate cancer cell lines DU145 and PC-3 resulted in sensitivity to apoptosis and inhibition of cell motility and invasion, whereas cell proliferation were not affected. We identified myristoylated alanine-rich protein kinase c substrate (MARCKS), which plays key roles in cell motility, as a new target in prostate cancer cells. Our data suggested that miR-21 could promote apoptosis resistance, motility, and invasion in prostate cancer cells and these effects of miR-21 may be partly due to its regulation of PDCD4, TPM1, and MARCKS. Gene therapy using miR-21 inhibition strategy may therefore be useful as a prostate cancer therapy. PMID- 19302978 TI - CpG islands: algorithms and applications in methylation studies. AB - Methylation occurs frequently at 5'-cytosine of the CpG dinucleotides in vertebrate genomes; however, this epigenetic feature is rarely observed in CpG islands (CGIs) or CpG clusters in the promoter regions of genes. Aberrant methylation of the promoter-associated CGIs might influence gene expression and cause carcinogenesis. Because of the functional importance, multiple algorithms have been available for identifying CGIs in a genome or a sequence. They can be categorized into the traditional algorithms (e.g., Gardiner-Garden and Frommer (1987), Takai and Jones (2002), and CpGPRoD (2002)) or statistical property based algorithms (CpGcluster (2006) and CG cluster (2007)). We reviewed the features of these algorithms and evaluated their performance on identifying functional CGIs using genome-wide methylation data. Moreover, identification of CGIs is an initial step in many recent studies for predicting methylation status as well as in the design of methylation detection platforms. We reviewed the benchmarks and features used in these studies. PMID- 19302979 TI - O-GlcNAcylation of Sp1 interrupts Sp1 interaction with NF-Y. AB - O-linked N-acetylglucosamine (O-GlcNAc), a monosaccharide N-acetylglucosamine addition on nucleocytoplasmic proteins, is abundant in transcription regulators and has been implicated in gene regulation. Sp1 transcription factor is multiply modified by O-GlcNAc within its serine/threonine-rich region and glutamine-rich transactivation domain. In the present study, we show that O-GlcNAc of Sp1 serine/threonine-rich region interrupts a physical interaction between Sp1 and NF YA, thus inhibiting Sp1-NF-Y cooperative activation of gene transcription. Our results strengthen the notion that O-GlcNAc regulates gene transcription by modulating the protein-protein interaction network among transcription regulatory proteins. PMID- 19302980 TI - The radial glia antibody RC2 recognizes a protein encoded by Nestin. AB - The RC2 antibody is widely used to label mouse radial glial cells in the developing central nervous system. While the antibody is known to recognize a 295 kDa intermediate filament proximal protein, the gene encoding the RC2 antigen remains to be identified. Here, we present evidences clearly demonstrating that Nestin encodes the RC2 antigen. First, the RC2 antigen and nestin have the same molecular weight and very similar tissue distribution. Second, genetic manipulations altering nestin expression also exert the same effect on the expression of the RC2 antigen. In particular, Nestin null mutation completely abolishes the RC2 immunoreactivity. Third, the expression of a truncated mouse nestin in Nestin-/- cells produces a small RC2 antigen whose size is the same to that of the truncated nestin. Furthermore, our data suggest that the RC2 antibody recognizes the C-terminal domain of nestin with unidentified posttranslational modification(s). PMID- 19302981 TI - Neuroplasticity and neuroprotection in enteric neurons: role of epithelial cells. AB - Neurons of enteric nervous system (ENS) regulate intestinal epithelial cells (IEC) functions but whether IEC can impact upon the neurochemical coding and survival of enteric neurons remain unknown. Neuro-epithelial interactions were studied using a coculture model composed of IEC lines and primary culture of rat ENS or human neuroblastoma cells (SH-SY5Y). Neurochemical coding of enteric neurons was analysed by immunohistochemistry and quantitative PCR. Neuroprotective effects of IEC were tested by measuring neuron specific enolase (NSE) release or cell permeability to 7-amino-actinomycin D (7-AAD). Following coculture with IEC, the percentage of VIP-immunoreactive (IR) neurons but not NOS IR and VIP mRNA expression were significantly increased. IEC significantly reduced dopamine-induced NSE release and 7-AAD permeability in culture of ENS and SH-SY5Y, respectively. Finally, we showed that NGF had neuroprotective effects but reduced VIP expression in enteric neurons. In conclusion, our study identified a novel role for IEC in the regulation of enteric neuronal properties. PMID- 19302982 TI - HBx protein induces EMT through c-Src activation in SMMC-7721 hepatoma cell line. AB - The relationships between epithelial-to-mesenchymal transition (EMT), hepatitis B virus X protein (HBx), and the non-receptor tyrosine kinase c-Src were investigated. The HBx gene transfected SMMC-7721 cells underwent morphological changes from a classic epithelial morphology to a spindle-like shape. The HBx transfection increased the invasive potential of these cells. When the transfected cells were exposed to the c-Src kinase inhibitor PP2, the cells recovered their original epithelial morphology and the mRNA and protein expression of epithelial and mesenchymal markers returned to the parental cell levels. Our data suggest that activated c-Src played a critical role in the HBx induced EMT of SMMC-7721 cells. PMID- 19302983 TI - Refined anatomical isolation of functional sleep circuits exhibits distinctive regional and circadian gene transcriptional profiles. AB - Powerful new approaches to study molecular variation in distinct neuronal populations have recently been developed enabling a more precise investigation of the control of neural circuits involved in complex behaviors such as wake and sleep. We applied laser capture microdissection (LCM) to isolate precise brain nuclei from rat CNS at opposing circadian time points associated with wake and sleep. Discrete anatomical and temporal analysis was performed to examine the extent of variation in the transcriptional control associated with both identifiable anatomical nuclei and with light/dark cycle. Precise isolation of specific brain nuclei regulating sleep and arousal, including the LC, SCN, TMN, VTA, and VLPO, demonstrated robust changes in gene expression. Many of these differences were not observed in previous studies where whole brain lysates or gross dissections were used to probe for changes in gene expression. The robust and differential profiles of genomic data obtained from the approaches used herein underscore the requirement for careful anatomical refinement in CNS gene expression studies designed to understand genomic control within behaviorally linked, but functionally isolated brain nuclei. PMID- 19302984 TI - Peripheral sensory deafferentation affects olfactory bulb neurogenesis in zebrafish. AB - The potential effects of the removal of olfactory input on adult neurogenesis in the olfactory bulb were examined. Olfactory organs of adult zebrafish were permanently and completely ablated by cautery and animals were exposed to bromodeoxyuridine then examined following short (4-hour) or long (3-week) survival periods. Short survival times allowed analysis of cell proliferation in the olfactory bulb. Long survival times permitted investigation of survival of adult-formed cells. Deafferentation did not immediately affect the dividing cells in the bulb but did affect the number of adult-formed cells, some of which expressed a neuronal marker, present in the bulb 3 weeks later. Thus, afferent removal influenced the fate of newly formed cells by impacting subsequent divisions, maturation, or survival of those cells. One week of deafferentation altered the pattern of cell genesis, with a significant increase in the number of dividing cells located in the olfactory bulb and also in the ventral telencephalic proliferation zone. Sham surgery did not impact either proliferation or survival of adult-formed cells in the olfactory bulb, suggesting that the deafferentation effect is specific. Thus, afferent innervation is necessary for normal cell proliferation and maintenance of the olfactory bulb in adult zebrafish. PMID- 19302985 TI - Stimulus pattern dependence of the Alzheimer's disease amyloid-beta 42 peptide's inhibition of long term potentiation in mouse hippocampal slices. AB - Increasing evidence has pointed to inhibition of Long Term Potentiation (LTP) by soluble A beta 42 oligomers as central in the etiology of the learning and memory deficits that are hallmarks of Alzheimer Disease. These effects are thought to occur by an interaction between A beta 42 and certain cellular effectors that induce LTP, however, the precise identity of the A beta 42-interactive signaling molecules is unknown. Identification of such effectors is made more difficult because LTP induced by different stimulation protocols can be expressed through heterogeneous signaling pathways. The aim of this study was to compare differences in the A beta 42-dependent levels of inhibition of LTPs that were induced using high frequency stimulation (HFS), versus theta burst stimulation (TBS). Our results show that untreated control brain slices tetanized with either HFS or TBS gave similar levels of LTP and post tetanic stimulation (PTP), suggesting that the response induced by either protocol was comparable. However, A beta 42 peptide significantly blocked LTP and PTP induced by HFS, but not when TBS was used. NMDA receptor antagonists, D-AP5 and ifenprodil, both blocked LTPs that were induced by HFS or TBS. We propose that unknown signaling effectors, other than the NMDA receptor, which are differentially involved in the induction of LTP by TBS, as compared to HFS, may be responsible for this resistance of TBS induced LTP to A beta 42 dependent inhibition. PMID- 19302987 TI - Morphological definition of CD71 positive reticulocytes by various staining techniques and electron microscopy compared to reticulocytes detected by an automated hematology analyzer. AB - BACKGROUND: The enumeration of peripheral blood reticulocytes plays an important part in clinical hematology. Although reticulocyte enumeration is currently performed with visible dyes such as New Methylene Blue (NMB), fluorescent dyes, or anti-CD71 (transferrin receptor) antibody, it has not been demonstrated whether the reticulocytes detected in each method are the same or not. METHODS: We prepared the reticulocyte rich fraction with density gradient centrifugation, stained with both anti-CD71 and Sysmex's fluorescent stain RET SEARCH (II), and detected the cells by both confocal laser scanning microscopy and flow cytometry. We also stained the reticulocyte rich fraction and the CD71+ reticulocytes with NMB and compared them by microscopy. We also observed the CD71+ reticulocytes by electron microscopy. RESULTS: Almost all CD71+ reticulocytes were intensely stained with both NMB and RET SEARCH (II). These cells were therefore classified as highly immature reticulocytes. During the stages of reticulocyte maturation, the expression of CD71 antigen decreased prior to the reduction of the reticular structures. The electron microscopic observation showed that CD71+ reticulocytes had some typical morphological characteristics found in highly immature reticulocytes. CONCLUSIONS: The CD71+ reticulocytes consisted of highly immature reticulocytes and were not equal to the reticulocytes defined with NMB or RET SEARCH (II). PMID- 19302986 TI - Neuroprotective effect of adenoviral catalase gene transfer in cortical neuronal cultures. AB - Reduced availability of reactive oxygen species is a key component of neuroprotection against various toxic stimuli. Recently we showed that the hydrogen peroxide scavenger catalase plays a central role in delayed preconditioning induced by the mitochondrial ATP-sensitive potassium channel opener BMS-191095. The purpose of the experiments discussed here was to investigate the neuroprotective effect of catalase in vitro using a recombinant adenoviral catalase gene transfer protocol. To induce catalase overexpression, cultured rat cortical neurons were infected with the adenoviral vector Ad5CMVcatalase and control cells were incubated with Ad5CMVntLacZ for 24 h. Gene transfer effectively increased catalase protein levels and activity, but did not influence other antioxidants tested. Ad5CMVcatalase, with up to 10 plaque forming units (pfu) per neuron, did not affect cell viability under control conditions and did not protect against glutamate excitotoxicity or oxygen-glucose deprivation. In contrast, catalase overexpression conferred a dose-dependent protection against exposure to hydrogen peroxide (viability: control, 33.02+/ 1.09%; LacZ 10 pfu/cell, 32.85+/-1.51%; catalase 1 pfu/cell, 62.09+/-4.17%*; catalase 2 pfu/cell, 98.71+/-3.35%*; catalase 10 pfu/cell, 99.68+/-1.99%*; *p<0.05 vs. control; mean+/-SEM). Finally, the protection could be antagonized using the catalase inhibitor 3-aminotriazole. Our results support the view that enhancing cellular antioxidant capacity may play a crucial role in neuroprotective strategies. PMID- 19302988 TI - Pre-analytical workstations: a tool for reducing laboratory errors. AB - Laboratory testing, a highly complex process commonly called the total testing process (TTP), is usually subdivided into three traditional (pre-, intra-, and post-) analytical phases. The majority of errors in TTP originate in the pre analytical phase, being due to individual or system design defects. In order to reduce errors in TTP, the pre-analytical phase should therefore be prioritized. In addition to developing procedures, providing training, improving interdepartmental cooperation, information technology and robotics may be a tool to reduce errors in specimen collection and pre-analytical sample handling. It has been estimated that >2000 clinical laboratories worldwide use total or subtotal automation supporting pre-analytic activities, with a high rate of increase compared to 2007; the need to reduce errors seems to be the catalyst for increasing the use of robotics. Automated systems to prevent medical personnel from drawing blood from the wrong patient were introduced commercially in the early 1990s. Correct patient identification and test tube labelling before phlebotomy are of extreme importance for patient safety in TTP, but currently few laboratories are interested in such products. At San Bassiano hospital, the implementation of advanced information technology and robotics in the pre analytical phase (specimen collection and pre-analytical sample handling) have improved accuracy, and clinical efficiency of the laboratory process and created a TTP that minimizes errors. PMID- 19302989 TI - Errors in medicine. AB - Modern awareness of the problem of medical injury--complications of treatment- can be fairly dated to the publication in 1991 of the results of the Harvard Medical Practice Study, but it was not until the publication of the 2000 Institute of Medicine (IOM) report, To Err is Human that patient safety really came to medical and public attention. Medical injury is a serious problem, affecting, as multiple studies have now shown, approximately 10% of hospitalized patients, and causing hundreds of thousands of preventable deaths each year. The organizing principle is that the cause is not bad people, it is bad systems. This concept is transforming; it replaces the previous exclusive focus on individual error with a focus on defective systems. Although the major focus on patient safety has been on implementing safe practices, it has become increasingly apparent that achieving a high level of safety in our health care organizations requires much more: several streams have emerged. One of these is the recognition of the importance of engaging patients more fully in their care. Another is the need for transparency. In the current health care organizational environment in most hospitals, at least six major changes are required to begin the journey to a culture of safety: 1. We need to move from looking at errors as individual failures to realizing they are caused by system failures; 2. We must move from a punitive environment to a just culture; 3. We move from secrecy to transparency; 4. Care changes from being provider (doctors) centered to being patient-centered; 5. We move our models of care from reliance on independent, individual performance excellence to interdependent, collaborative, interprofessional teamwork; 6. Accountability is universal and reciprocal, not top-down. PMID- 19302990 TI - Proteome analysis of gelatin-bound salivary proteins in patients with primary Sjogren's syndrome: identification of matrix metalloproteinase-9. PMID- 19302991 TI - Helicase dependent OnChip-amplification and its use in multiplex pathogen detection. AB - BACKGROUND: The need for fast, specific and sensitive multiparametric detection methods is an ever growing demand in molecular diagnostics. Here we report on a newly developed method, the helicase dependent OnChip amplification (OnChip-HDA). This approach integrates the analysis and detection in one single reaction thus leading to time and cost savings in multiparametric analysis. METHODS: HDA is an isothermal amplification method that is not depending on thermocycling as known from PCR due to the helicases' ability to unwind DNA double-strands. We have combined the HDA with microarray based detection, making it suitable for multiplex detection. As an example we used the OnChip HDA in single and multiplex amplifications for the detection of the two pathogens N. gonorrhoeae and S. aureus directly on surface bound primers. RESULTS: We have successfully shown the OnChip-HDA and applied it for single- and duplex-detection of the pathogens N. gonorrhoeae and S. aureus. CONCLUSION: We have developed a new method, the OnChip HDA for the multiplex detection of pathogens. Its simplicity in reaction setup and potential for miniaturization and multiparametric analysis is advantageous for the integration in miniaturized Lab on Chip systems, e.g. needed in point of care diagnostics. PMID- 19302992 TI - Laboratory safety and the WHO World Alliance for Patient Safety. AB - Laboratory medicine has been a pioneer in the field of patient safety; indeed, the College of American Pathology first called attention to the issue in 1946. Delivering reliable laboratory results has long been considered a priority, as the data produced in laboratory medicine have the potential to critically influence individual patients' diagnosis and management. Until recently, most attention on laboratory safety has focused on the analytic stage of laboratory medicine. Addressing this stage has led to significant and impressive improvements in the areas over which laboratories have direct control. However, recent data demonstrate that pre- and post-analytical phases are at least as vulnerable to errors; to further improve patient safety in laboratory medicine, attention must now be focused on the pre- and post-analytic phases, and the concept of patient safety as a multi-disciplinary, multi-stage and multi-system concept better understood. The World Alliance for Patient Safety (WAPS) supports improvement of patient safety globally and provides a potential framework for considering the total testing process. PMID- 19302993 TI - Governance of preanalytical variability: travelling the right path to the bright side of the moon? AB - Medical errors can be traditionally clustered into 4 categories, which include errors of diagnosis, errors of treatment, errors of prevention, and an 'other miscellaneous' category. Owing to the volume and complexity of testing, and considering that laboratory error is defined as any defect from ordering tests to reporting results and appropriately interpreting and reacting on these, it is not surprising that mistakes in the total testing process occur with frequency, have connections to all four types of medical errors and represent a serious hazard for patient health. Throughout the laboratory diagnostics, preanalytical problems prevail. Moreover, the positive trends towards reduction of laboratory errors over the past decade, particularly those in the analytical phase, has little involved the preanalytical phase, which actually represents the most critical area to target. In particular, the high frequency of errors still attributable to processes external to the laboratory requires additional efforts for the governance of this mistreated phase of the total testing process, so that we can finally find the right path to progress from the dark to the bright side of the moon. As for any other type of medical errors, the most effective path to improvement is the implementation of a total quality management system, encompassing a multifaceted strategy for process and risk analysis, based on error prevention, detection, and management. PMID- 19302994 TI - Prevalence of serum antibodies to TORCH among women before pregnancy or in the early period of pregnancy in Beijing. AB - BACKGROUND: The primary infection of Toxoplasma gondii (TOX), rubella virus (RV), cytomegalovirus (CMV), or herpes simplex virus (HSV), abbreviated as TORCH, remains a major problem in the pregnant women in China today. METHODS: We analyzed the performance of a novel chemiluminesent immunoassay (CLIA) for anti TORCH IgM and IgG detection then analyzed the prevalence of TORCH infection among 4692 women at childbearing age in Beijing, China. RESULTS: The functional sensitivity of detecting anti-TOX IgM, anti-RV IgM, anti-CMV IgM and anti-HSV (types 1 and 2) IgM were 1.4, 3.2, 1.0 AU/ml and 0.5 index, respectively. The within-assay and the total CVs were both <10%, and recoveries of these assays ranged from 90-110%. High concentration of hemoglobin, lipids and bilirubin in sample did not affect the results. The infective rate of TORCH was 17.2%, with the highest positive rate of anti-HSV IgM. Within anti-TORCH IgG, anti-CMV IgG had the highest infective rate, 92.5%. CONCLUSIONS: The infective rate of TORCH was fairly high among women before pregnancy or in the early period of pregnancy in Beijing. Entirely new approaches to prevention and treatment of congenital TORCH infection are necessary. PMID- 19302995 TI - Exploring the iceberg of errors in laboratory medicine. AB - The last few decades have seen a significant decrease in the rates of analytical errors in clinical laboratories, and currently available evidence demonstrates that the pre- and post-analytical steps of the total testing process (TTP) are more error-prone than the analytical phase. In particular, most errors are identified in pre-pre-analytic and post-post analytic steps outside the walls of the laboratory, and beyond its control. However, in a patient-centered approach to the delivery of health care services, there is the need to investigate any possible defect in the total testing process that may have a negative impact on the patient. In fact, in the interests of patients, any direct or indirect negative consequence related to a laboratory test must be considered, irrespective of which step is involved and whether the error is caused by a laboratory professional (e.g., calibration or testing error) or by a non laboratory operator (e.g., inappropriate test request, error in patient identification and/or blood collection). Data on diagnostic errors in primary care and in the emergency department setting demonstrate that inappropriate test requesting and incorrect interpretation account for a large percentage of total errors whatever the discipline involved, be it radiology, pathology or laboratory medicine. Patient misidentification and problems in communicating results, which affect the delivery of all diagnostic services, are widely recognized as the main goals for quality improvement. Therefore, some common problems affect diagnostic errors, although specific faults characterising errors in laboratory medicine should lead to preventive and corrective actions if evidence-based quality indicators are developed, implemented and monitored. The lesson we have learned is that each practice must examine its own total testing process to discover its weaknesses and identify appropriate remedies. PMID- 19302996 TI - The reporting, classification and grading of quality failures in the medical laboratory. AB - Laboratory errors have a reported frequency of 0.012 to 0.6% of all test results and given the central role of the laboratory in diagnosis, may have a major adverse impact on patient care. Laboratories have been at the forefront of efforts to enhance patient safety through a range of improvements such as increased automation of manual processes and analytical quality control programmes. It is important that all laboratories have active systems in place to identify and monitor quality failures. This will be facilitated by a systems based approach to error which seeks to identify and correct weaknesses in policies/procedures rather than to apportion blame. Quality failures may be classified by cause [i.e., step in the testing pathway where the problem occurred] and graded by severity on a 5 point scale. The severity grading score measures both the Actual impact ['A' score] on patient outcome and the Potential impact ['P' score] i.e., the worst case possible outcome that might have resulted. 'A' scores tend to be skewed towards low adverse patient impact while 'P' scores are skewed towards high adverse impact. This confirms the need for ongoing vigilance by laboratories. This classification/grading system is easy to implement and represents a valuable additional tool for monitoring performance. PMID- 19302997 TI - Serum mesothelin has a higher diagnostic utility than hyaluronic acid in malignant mesothelioma. AB - We assessed comparatively the diagnostic value of two potential malignant pleural mesothelioma (MPM) markers: hyaluronic acid (HA) and soluble mesothelin. MATERIALS AND METHOD: We measured serum and pleural fluid values of mesothelin and hyaluronic acid in 76 patients with MPM, 33 patients with pleural metastases of carcinomas (Mets group) and 27 patients with benign pleural effusion related to asbestos exposure (BPLAE). RESULTS: Using a serum HA cut-off of 100 microg/L, 8 patients/33 (24.2%) were positive in the Mets group versus 20/76 (26.3%) in the MPM group and only 1/27 BPLAE patients. The area under ROC curve for serum HA in MPM versus Mets or BPLAE groups was only 0.617 while it was 0.755 for mesothelin. In pleural fluid, both markers had similar diagnostic values. CONCLUSIONS: Serum mesothelin is more sensitive than hyaluronic acid in diagnosing MPM and there is no benefit in combining both markers. PMID- 19302998 TI - On the importance of the use of proper approaches for comparison of analytical methods for serum nitrate and for evaluation of reference concentrations. PMID- 19302999 TI - Common genetic variants of the vitamin D binding protein (DBP) predict differences in response of serum 25-hydroxyvitamin D [25(OH)D] to vitamin D supplementation. AB - BACKGROUND: To determine the effect of vitamin D binding protein (DBP) genotypes on 25-hydroxyvitamin D [25(OH)D] changes with vitamin D supplements, we studied 98 adults receiving 600 or 4000 IU/d vitamin D(3) for one year. METHODS: The DBP functional variant, T436K, was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Mean 25(OH)D increases were 97% for TT (n=48), 151% for TK (n=31) and 307% (n=6) for KK genotypes (p=.004). CONCLUSIONS: As with baseline 25(OH)D, T436K genotype predicts 25(OH)D changes after long-term vitamin D supplementation. PMID- 19303000 TI - Serum retinol binding protein 4 in patients with familial partial lipodystrophy. AB - OBJECTIVE: To determine Retinol Binding Protein 4 (RBP4) levels in patients with Familial Partial Lipodystrophy (FPLD). METHODS: Ten patients with FPLD and a control group (9 patients) were selected to participate in the study. RESULTS: RBP4-log levels were lower in patients with FPLD in comparison to control group (1.52 +/- 0.32 vs 1.84+/-0.25, p=0.029). A statistical trend was observed between Waist-to-Hip Ratio and RBP4-log (r=-0.44, p=0.054). CONCLUSION: RBP4 levels are decreased in FPLD. PMID- 19303001 TI - Bidis--hand-rolled, Indian cigarettes: induced biochemical changes in plasma and red cell membranes of human male volunteers. AB - OBJECTIVE: To investigate the effect of bidi smoking on erythrocyte antioxidant status, membrane fluidity. DESIGN AND METHODS: Thirty experimental and control subjects (mean age 35+/-5) were selected for the study. Experimental subjects smoke 22+/-4 bidis per day for 8-10 years. RESULTS: Increase in plasma total cholesterol, LDL-cholesterol, triglycerides, lipid peroxidation, protein carbonyls with a decrease in HDL-cholesterol, thiol groups as well as increased erythrocyte catalase (CAT), superoxide dismutase (SOD), decreased glutathione peroxidase (GPx) activity and reduced glutathione (GSH) content was observed in bidi smokers. Increase in the erythrocyte membrane lipid peroxidation, cholesterol phospholipids (C/P) ratio as well as decrease in protein and Na(+)/K(+)-ATPase activity was observed. Increase in nitrite/nitrate (NOx) levels of plasma, red cell lysate was positively correlated with C/P ratio (r=0.614) and Na(+)/K(+)-ATPase (r=0.435) in bidi smokers. CONCLUSIONS: Bidi smoke alters antioxidant status, red cell membrane fluidity and increases atherogenicity. PMID- 19303002 TI - Syndecan-1 regulates BMP signaling and dorso-ventral patterning of the ectoderm during early Xenopus development. AB - Extracellular regulation of growth factor signaling is a key event for embryonic patterning. Heparan sulfate proteoglycans (HSPG) are among the molecules that regulate this signaling during embryonic development. Here we study the function of syndecan1 (Syn1), a cell-surface HSPG expressed in the non-neural ectoderm during early development of Xenopus embryos. Overexpression of Xenopus Syn1 (xSyn1) mRNA is sufficient to reduce BMP signaling, induce chordin expression and rescue dorso-ventral patterning in ventralized embryos. Experiments using chordin morpholinos established that xSyn1 mRNA can inhibit BMP signaling in the absence of chordin. Knockdown of xSyn1 resulted in a reduction of BMP signaling and expansion of the neural plate with the concomitant reduction of the non-neural ectoderm. Overexpression of xSyn1 mRNA in xSyn1 morphant embryos resulted in a biphasic effect, with BMP being inhibited at high concentrations and activated at low concentrations of xSyn1. Interestingly, the function of xSyn1 on dorso ventral patterning and BMP signaling is specific for this HSPG. In summary, we report that xSyn1 regulates dorso-ventral patterning of the ectoderm through modulation of BMP signaling. PMID- 19303003 TI - NAT10, a nucleolar protein, localizes to the midbody and regulates cytokinesis and acetylation of microtubules. AB - The midbody is a structural organelle formed in late phase mitosis which is responsible for completion of cytokinesis. Although various kinds of proteins have been found to distribute or immigrate to this organelle, their functions have still not been completely worked out. In this study, we demonstrated that NAT10 (N-acetyltransferase 10, NAT10) is not only predominantly distributed in the nucleolus in interphase, but is also concentrated in the mitotic midbody during telophase. The domain in N-terminal residues 549-834 of NAT10 specifically mediated its subcellular localization. Treatment with genotoxic agents or irradiation increased concentration of NAT10 in both the nucleolus and midbody. Moreover, DNA damage induced increase of NAT10 in the midbody apparently accompanied by in situ elevation of the level of acetylated alpha-tubulin, suggesting that it plays a role in maintaining or enhancing stability of alpha tubulin. The depletion of NAT10 induced defects in nucleolar assembly, cytokinesis and decreased acetylated alpha-tubulin, leading to G2/M cell cycle arrest or delay of mitotic exit. In addition, over-expression of NAT10 was found in a variety of soft tissue sarcomas, and correlated with tumor histological grading. These results indicate that NAT10 may play an important role in cell division through facilitating reformation of the nucleolus and midbody in the late phase of cell mitosis, and stabilization of microtubules. PMID- 19303004 TI - Clinical and genetic features of TGFBI-linked corneal dystrophies in Mexican population: description of novel mutations and novel genotype-phenotype correlations. AB - Corneal dystrophies (CDS) are inherited disorders characterized by an altered corneal transparency and refractive index which may be caused by a progressive accumulation of deposits within the different corneal layers. Most CDs are inherited in an autosomal dominant fashion and mutations in the TGFBI gene at chromosome 5q31 cause the majority of CDs affecting the stromal layer. A genotype phenotype correlation has been identified in most analyzed populations as specific amino acid changes in TGFBI protein cause specific stromal phenotypes. However, analysis of additional populations will help to broaden the mutational spectrum ultimately allowing a better clinical-molecular classification of patients with this group of diseases. In this work, eighteen unrelated Mexican probands suffering from stromal CDs were clinically assessed and their TGFBI gene status investigated. Complete ophthalmologic evaluation, including biomicroscopic inspection and dilated fundus examination, was performed. In addition, detailed genealogical analyses as well as automated DNA sequencing of the entire TGFBI gene were done in all probands. Mutation-carrying exons were examined in 50 first and second degree relatives. Phenotypic analysis disclosed the occurrence of 6 cases of lattice CD, 6 of granular CD, 2 of granular type 2 (Avellino CD), 2 of polymorphic corneal amyloidosis, 1 of Reis-Bucklers CD, and 1 of an unclassifiable phenotype. TGFBI mutations were identified in all 18 probands. A total of six different mutations were observed: p.V113I, p.M502V, p.A546D, p.L550P, p.R555W, and p.H626R. Of these, mutations p.L550P (originated by the change c.1649T>C at exon 12), p.M502V (c.1504A>G, at exon 11), and p.V113I (c.337G>A, at exon 4), are novel TGFBI mutations. All subjects with lattice CD in our sample carried the p.H626R mutation. No instances of defects at codon 124, one of the two most frequently mutated sites in TGFBI-linked CDs, were detected. A distinct TGFBI mutational pattern was identified in Mexican patients with stromal CDs. Novel TGFBI mutations and new genotype-phenotype correlations were also recognized. This study stresses the importance of performing TGFBI genetic analysis in distinct CD populations. PMID- 19303005 TI - Impaired mitochondrial dynamics and function in the pathogenesis of Parkinson's disease. AB - Parkinson's disease (PD), the most frequent movement disorder, is caused by the progressive loss of the dopamine neurons within the substantia nigra pars compacta (SNc) and the associated deficiency of the neurotransmitter dopamine in the striatum. Most cases of PD occur sporadically with unknown cause, but mutations in several genes have been linked to genetic forms of PD (alpha synuclein, Parkin, DJ-1, PINK1, and LRRK2). These genes have provided exciting new avenues to study PD pathogenesis and the mechanisms underlying the selective dopaminergic neuron death in PD. Epidemiological studies in humans, as well as molecular studies in toxin-induced and genetic animal models of PD show that mitochondrial dysfunction is a defect occurring early in the pathogenesis of both sporadic and familial PD. Mitochondrial dynamics (fission, fusion, migration) is important for neurotransmission, synaptic maintenance and neuronal survival. Recent studies have shown that PINK1 and Parkin play crucial roles in the regulation of mitochondrial dynamics and function. Mutations in DJ-1 and Parkin render animals more susceptible to oxidative stress and mitochondrial toxins implicated in sporadic PD, lending support to the hypothesis that some PD cases may be caused by gene-environmental factor interactions. A small proportion of alpha-synuclein is imported into mitochondria, where it accumulates in the brains of PD patients and may impair respiratory complex I activity. Accumulation of clonal, somatic mitochondrial DNA deletions has been observed in the substantia nigra during aging and in PD, suggesting that mitochondrial DNA mutations in some instances may pre-dispose to dopamine neuron death by impairing respiration. Besides compromising cellular energy production, mitochondrial dysfunction is associated with the generation of oxidative stress, and dysfunctional mitochondria more readily mediate the induction of apoptosis, especially in the face of cellular stress. Collectively, the studies examined and summarized here reveal an important causal role for mitochondrial dysfunction in PD pathogenesis, and suggest that drugs and genetic approaches with the ability to modulate mitochondrial dynamics, function and biogenesis may have important clinical applications in the future treatment of PD. PMID- 19303006 TI - Partial inhibition of proteasome activity enhances remyelination after cuprizone induced demyelination. AB - We have previously demonstrated that addition of low concentrations of lactacystin (a specific inhibitor of the proteasome) to oligodendroglial cell cultures containing a high percentage of precursor cells induces their exit from the cell cycle and their differentiation. On the other hand, we have recently shown that the mechanism of cuprizone toxicity on oligodendroglial cells involves the recruitment of microglia and their secretion of pro-inflammatory cytokines and in the increased production of oxidant species, which results in a decrease in the activities of the mitochondrial respiratory chain. In the present paper we investigated the effect of a decrease in proteasome activity induced by the injection of lactacystin in the corpus callosum in the remyelination process that normally occurs after cuprizone-induced demyelination. This treatment markedly improves the remyelination process that normally occurs in cuprizone-induced demyelination. It also attenuates the activation of NFkappaB and the recruitment of microglia and astrocytes, thus helping in the recovery of the mitochondrial respiratory chain activities that are affected by cuprizone treatment. PMID- 19303007 TI - Pharmacological inhibition of mitochondrial membrane permeabilization for neuroprotection. AB - Recent data have provided important clues about the molecular mechanisms underlying certain neurodegenerative diseases. Most cell death in vertebrates proceeds via the mitochondrial pathway of apoptosis. Mitochondria contain proapoptotic factors such as cytochrome c and AIF in their intermembrane space. Furthermore, mitochondrial membrane permeabilization (MMP) is a critical event during apoptosis, representing the "point of no return" of the lethal process. Modern medicine is developing an increasing number of drugs for neurodegenerative disease, but no neuroprotective treatment has yet been established. While current treatments temporarily alleviate symptoms, they do not halt disease progression. This paper briefly reviews the pharmacological inhibition of mitochondrial membrane permeabilization for neuroprotection. PMID- 19303008 TI - Trypanosoma rangeli: a possible role for ecto-phosphatase activity on cell proliferation. AB - Here we demonstrate for the first time that growth of Trypanosoma rangeli, a protozoa parasite, is strongly dependent on the presence of inorganic phosphate (Pi) in the culture medium and that the replacement of the inorganic phosphate in the culture medium by beta-glycerophosphate, a substrate for phosphatases lead the cells to achieve its maximal growth. The ecto-phosphatase activity present on the external surface of T. rangeli decreased during the growth phase of the parasite, suggesting that this enzyme could be important for the development. Accordingly, the inhibition of this ecto-phosphatase activity by sodium orthovanadate also inhibited the proliferation of T. rangeli. Parasites maintained in a Pi-starved culture medium (2 mM Pi) had 4-fold more ecto phosphatase activity as compared to parasites maintained in a Pi-supplemented culture medium (50 mM Pi). Altogether, these results presented here suggest that this ecto-phosphatase activity leads to hydrolysis of phosphorylated compounds present in the extracellular medium, which could contribute to the acquisition of inorganic phosphate during the development of T. rangeli epimastigotes. PMID- 19303009 TI - Taxonomy and species delimitation in Cryptosporidium. AB - Amphibians, reptiles, birds and mammals serve as hosts for 19 species of Cryptosporidium. All 19 species have been confirmed by morphological, biological, and molecular data. Fish serve as hosts for three additional species, all of which lack supporting molecular data. In addition to the named species, gene sequence data from more than 40 isolates from various vertebrate hosts are reported in the scientific literature or are listed in GenBank. These isolates lack taxonomic status and are referred to as genotypes based on the host of origin. Undoubtedly, some will eventually be recognized as species. For them to receive taxonomic status sufficient morphological, biological, and molecular data are required and names must comply with the rules of the International Code for Zoological Nomenclature (ICZN). Because the ICZN rules may be interpreted differently by persons proposing names, original names might be improperly assigned, original literature might be overlooked, or new scientific methods might be applicable to determining taxonomic status, the names of species and higher taxa are not immutable. The rapidly evolving taxonomic status of Cryptosporidium sp. reflects these considerations. PMID- 19303010 TI - Prodigiosin is not a determinant factor in lysis of Leishmania (Viannia) braziliensis after interaction with Serratia marcescens D-mannose sensitive fimbriae. AB - In this paper, the lytic activity of two variants of Serratia marcescens against promastigotes of Leishmania braziliensis was studied. In vitro assays showed that S. marcescens variant SM365 lyses L. braziliensis promastigotes, while the variant DB11 did not. Scanning electron microscopy (SEM) revealed that S. marcescens SM365 adheres to all cellular body and flagellum of the parasite. Several filamentous structures were formed and identified as biofilms. After 120min incubation, they connect the protozoan to the developing bacterial clusters. SEM also demonstrated that bacteria, adhered onto L. braziliensis promastigote surface, formed small filamentous structures which apparently penetrates into the parasite membrane. d-mannose protects L. braziliensis against the S. marcescens SM365 lytic effect in a dose dependent manner. SM365 variant pre cultivated at 37 degrees C did not synthesize prodigiosin although the adherence and lysis of L. braziliensis were similar to the effect observed with bacteria cultivated at 28 degrees C, which produce high concentrations of prodigiosin. Thus, we suggest that prodigiosin is not involved in the lysis of promastigotes and that adherence promoted by bacterial mannose-sensitive (MS) fimbriae is a determinant factor in the lysis of L. braziliensis by S. marcescens SM365. PMID- 19303011 TI - The structural basis of allosteric regulation in proteins. AB - Allosteric regulation of protein function occurs when the regulatory trigger, such as the binding of a small-molecule effector or inhibitor, takes place some distance from the protein's, or protein complex's, active site. This distance can be a few A, or tens of A. Many proteins are regulated in this way and exhibit a variety of allosteric mechanisms. Here we review how analyses of experimentally determined models of protein 3D structures, using either X-ray crystallography or NMR spectroscopy, have revealed some of the mechanisms involved. PMID- 19303012 TI - Budesonide is effective in treating lymphocytic colitis: a randomized double blind placebo-controlled study. AB - BACKGROUND & AIMS: Budesonide is effective in treating collagenous colitis, but no treatment is established for lymphocytic colitis. We performed a randomized, double-blind, placebo-controlled study to evaluate the effects of budesonide in patients with lymphocytic colitis. METHODS: Forty-two patients (median age, 61 years) with lymphocytic colitis and chronic diarrhea were randomly assigned to groups that were given oral doses of budesonide (9 mg/d) or placebo for 6 weeks. Nonresponders at week 6 were given open-label budesonide (9 mg/d) for 6 additional weeks. A complete colonoscopy and histologic and quality-of-life analyses were performed at baseline and at week 6. The primary end point was clinical remission at 6 weeks, with last observation carried forward (LOCF). All patients who left the study in clinical remission were followed for relapse. RESULTS: At week 6, 86% of patients given budesonide were in clinical remission (with LOCF) compared with 48% of patients given placebo (P = .010). Furthermore, open-label budesonide therapy induced clinical remission in 7 of 8 patients given placebo. Histologic remission was observed in 73% of patients given budesonide compared with 31% given placebo (P = .030). Only 1 patient discontinued budesonide therapy prematurely. During a mean follow-up period of 14 months, 15 patients (44.1%) experienced a clinical relapse (after a mean of 2 months); 8 of the relapsing patients were retreated with and responded again to budesonide. CONCLUSIONS: Budesonide effectively induces clinical remission in patients with lymphocytic colitis and significantly improves histology results after 6 weeks. Clinical relapses occur but can be treated again with budesonide. PMID- 19303013 TI - Selection pressure from neutralizing antibodies drives sequence evolution during acute infection with hepatitis C virus. AB - BACKGROUND & AIMS: Despite recent characterization of hepatitis C virus-specific neutralizing antibodies, it is not clear to what extent immune pressure from neutralizing antibodies drives viral sequence evolution in vivo. This lack of understanding is particularly evident in acute infection, the phase when elimination or persistence of viral replication is determined and during which the importance of the humoral immune response has been largely discounted. METHODS: We analyzed envelope glycoprotein sequence evolution and neutralization of sequential autologous hepatitis C virus pseudoparticles in 8 individuals throughout acute infection. RESULTS: Amino acid substitutions occurred throughout the envelope genes, primarily within the hypervariable region 1 of E2. When individualized pseudoparticles expressing sequential envelope sequences were used to measure neutralization by autologous sera, antibodies neutralizing earlier sequence variants were detected at earlier time points than antibodies neutralizing later variants, indicating clearance and evolution of viral variants in response to pressure from neutralizing antibodies. To demonstrate the effects of amino acid substitution on neutralization, site-directed mutagenesis of a pseudoparticle envelope sequence revealed amino acid substitutions in hypervariable region 1 that were responsible for a dramatic decrease in neutralization sensitivity over time. In addition, high-titer neutralizing antibodies peaked at the time of viral clearance in all spontaneous resolvers, whereas chronically evolving subjects displayed low-titer or absent neutralizing antibodies throughout early acute infection. CONCLUSIONS: These findings indicate that, during acute hepatitis C virus infection in vivo, virus-specific neutralizing antibodies drive sequence evolution and, in some individuals, play a role in determining the outcome of infection. PMID- 19303014 TI - The role of the visceral mesoderm in the development of the gastrointestinal tract. AB - The gastrointestinal (GI) tract forms from the endoderm (which gives rise to the epithelium) and the mesoderm (which develops into the smooth muscle layer, the mesenchyme, and numerous other cell types). Much of what is known of GI development has been learned from studies of the endoderm and its derivatives, because of the importance of epithelial biology in understanding and treating human diseases. Although the necessity of epithelial-mesenchymal cross talk for GI development is uncontested, the role of the mesoderm remains comparatively less well understood. The transformation of the visceral mesoderm during development is remarkable; it differentiates from a very thin layer of cells into a complex tissue comprising smooth muscle cells, myofibroblasts, neurons, immune cells, endothelial cells, lymphatics, and extracellular matrix molecules, all contributing to the form and function of the digestive system. Understanding the molecular processes that govern the development of these cell types and elucidating their respective contribution to GI patterning could offer insight into the mechanisms that regulate cell fate decisions in the intestine, which has the unique property of rapid cell renewal for the maintenance of epithelial integrity. In reviewing evidence from both mammalian and nonmammalian models, we reveal the important role of the visceral mesoderm in the ontogeny of the GI tract. PMID- 19303015 TI - Angiotensin II activates I kappaB kinase phosphorylation of RelA at Ser 536 to promote myofibroblast survival and liver fibrosis. AB - BACKGROUND & AIMS: The transcription factor nuclear factor-kappaB (NF)-kappaB promotes survival of hepatic myofibroblasts and fibrogenesis through poorly defined mechanisms. We investigated the activities of angiotensin II and I kappaB kinase (IKK) in regulation of NF-kappaB activity and the role of these proteins in liver fibrosis in rodents and humans. METHODS: Phosphorylation of the NF kappaB subunit RelA at serine 536 (P-Ser(536)-RelA) was detected by immunoblot and immunohistochemical analyses. P-Ser(536)-RelA function was assessed using vectors that expressed mutant forms of RelA, cell-permeable blocking peptides, and assays for RelA nuclear transport and apoptosis. Levels of P-Ser(536)-RelA were compared with degree of fibrosis in liver sections from chronically injured rats and patients with hepatitis C virus-mediated fibrosis who had been treated with the AT1 antagonist losartan. RESULTS: Constitutive P-Ser(536)-RelA is a feature of human hepatic myofibroblasts, both in vitro and in situ in diseased livers. Autocrine angiotensin II stimulated IKK-mediated phosphorylation of RelA at Ser(536), which was required for nuclear transport and transcriptional activity of NF-kappaB. Inhibition of angiotensin II, the angiotensin II receptor type 1 (AT1), or IKK blocked Ser(536) phosphorylation and stimulated myofibroblast apoptosis. Treatment of fibrotic rodent liver with the angiotensin converting enzyme (ACE) inhibitor captopril or the IKK inhibitor sulphasalazine resulted in loss of P-Ser(536)-RelA-positive myofibroblasts and fibrosis regression. In human liver samples, increased numbers of P-Ser(536)-RelA-positive cells were associated with fibrosis that regressed following exposure to losartan. CONCLUSIONS: An autocrine pathway that includes angiotensin II, IKK, and P-Ser(536)-RelA regulates myofibroblast survival and can be targeted to stimulate therapeutic regression of liver fibrosis. PMID- 19303016 TI - P38 MAPK inhibitor semapimod reduces postoperative ileus via peripheral and central mechanisms. PMID- 19303017 TI - Hepatic stellate cells promote hepatocyte engraftment in rat liver after prostaglandin-endoperoxide synthase inhibition. AB - BACKGROUND & AIMS: Hepatic inflammation occurs immediately after cells are transplanted to the liver, but the mechanisms that underlie this process are not fully defined. We examined cyclooxygenase pathways that mediate hepatic inflammation through synthesis of prostaglandins, prostacyclins, thromboxanes, and other prostanoids following transplantation of hepatocytes. METHODS: We transplanted F344 rat hepatocytes into syngeneic dipeptidyl peptidase IV deficient F344 rats. Changes in cyclooxygenase pathways were analyzed, and specific pathways were blocked pharmacologically; the effects on cell engraftment and native liver cells were determined. RESULTS: Transplantation of hepatocytes induced hepatic expression of prostaglandin-endoperoxide synthases 1 and 2, which catalyze production of prostaglandin H2, as well as the downstream factor thromboxane synthase, which produces thromboxane A2 (a regulator of vascular and platelet responses in inflammation). Transplanted hepatocytes were in proximity with liver cells that expressed prostaglandin-endoperoxide synthases. The number of engrafted hepatocytes increased in rats given naproxen or celecoxib before transplantation but not in rats given furegrelate (an inhibitor of thromboxane synthase) or clopodigrel (an antiplatelet drug). Naproxen and celecoxib did not prevent hepatic ischemia or activation of neutrophils, Kupffer cells, or inflammatory cytokines, but they did induce hepatic stellate cells to express cytoprotective genes, vascular endothelial growth factor and hepatocyte growth factor, and matrix-type metalloproteinases and tissue inhibitor of metalloproteinase-1, which regulate hepatic remodeling. CONCLUSIONS: Activation of cyclooxygenase pathways interferes with engraftment of transplanted hepatocytes in the liver. Pharmacologic blockade of prostaglandin-endoperoxide synthases stimulated hepatic stellate cells and improved cell engraftment. PMID- 19303018 TI - Common cause of chronic diarrhea not to be forgotten. PMID- 19303019 TI - Frequent inactivation of axon guidance molecule RGMA in human colon cancer through genetic and epigenetic mechanisms. AB - BACKGROUND & AIMS: Repulsive guidance molecule member A (RGMA) is a glycosylphosphatidylinositol-anchored glycoprotein and axon guidance molecule that signals through its receptor, neogenin (NEO1), a homologue of the deleted-in colorectal cancer (DCC) gene. RGMA also functions as a bone morphogenetic protein (BMP) coreceptor. We studied the potential roles of RGMA and NEO1 in colorectal cancer (CRC) pathogenesis. METHODS: We analyzed expression of RGMA and NEO1, as well as their epigenetic and genetic changes, in a large series of CRC samples, normal colon tissues, adenomas, and cell lines. These studies were accompanied by in vitro functional assay. RESULTS: RGMA and NEO1 expression were significantly down-regulated in most CRCs, adenomas, and cell lines. RGMA was frequently silenced by promoter methylation in CRCs (86.7%), adenomas (90.9%), and CRC cell lines (92.3%) but not in normal colon tissues; allelic imbalance of RGMA and NEO1 was observed in 40% and 49% of CRCs, respectively. In CRC samples, reduced RGMA levels were significantly associated with mismatch repair deficiency or mutations in KRAS or BRAF. Exposure to 5-aza-2'-deoxycytidine restored RGMA expression in CRC cell lines. Transfection of RGMA into CRC cells suppressed cell proliferation, migration, and invasion and also increased apoptosis in response to DNA-damaging agent. CONCLUSIONS: The frequent genetic and epigenetic inactivation of RGMA in CRCs and adenomas along with its in vitro function collectively support its role as a tumor suppressor in colon cells. These findings add to the expanding list of axon guidance molecules with disrupted function during colon carcinogenesis and create new opportunities for early detection and drug development. PMID- 19303020 TI - Colony stimulating factor-1 dependence of paneth cell development in the mouse small intestine. AB - BACKGROUND & AIMS: Paneth cells (PCs) secrete defensins and antimicrobial enzymes that contribute to innate immunity against pathogen infections within the mucosa of the small intestine. We examined the role of colony stimulating factor-1 (CSF 1) in PC development. METHODS: CSF-1-deficient and CSF-1 receptor (CSF-1R) deficient mice and administration of neutralizing anti-CSF-1R antibody were used to study the requirement of CSF-1 for the development of epithelial cells of the small intestine. CSF-1 transgenic reporter mice and mice that express only the membrane-spanning, cell-surface CSF-1 isoform were used to investigate regulation by systemic versus local CSF-1. RESULTS: Mice deficient in CSF-1 or CSF-1R had greatly reduced numbers of mature PCs. PCs express the CSF-1R, and administration of anti-CSF-1R antibody to neonatal mice significantly reduced the number of PCs. Analysis of transgenic CSF-1 reporter mice showed that CSF-1-expressing cells are in close proximity to PCs. CSF-1/CSF-1R-deficient mice also had reduced numbers of the proliferating epithelial cell progenitors and lamina propria macrophages. Expression of the membrane-spanning, cell-surface CSF-1 isoform in CSF-1 deficient mice completely rescued the deficiencies of PCs, proliferating progenitors, and lamina propria macrophages. CONCLUSIONS: These results indicate local regulation by CSF-1 of PC development, either directly, in a juxtacrine/paracrine manner, or indirectly, by lamina propria macrophages. Therefore, CSF-1R hyperstimulation could be involved in hyperproliferative disorders of the small intestine, such as Crohn's disease and ulcerative colitis. PMID- 19303021 TI - Colitis-associated variant of TLR2 causes impaired mucosal repair because of TFF3 deficiency. AB - BACKGROUND & AIMS: Goblet cells (GC) facilitate mucosal protection and epithelial barrier repair, yet the innate immune mechanisms that selectively drive GC functions have not been defined. The aim of this study was to determine whether Toll-like receptor (TLR) 2 and modulation of GC-derived trefoil factor (TFF) 3 are functionally linked in the intestine. METHODS: GC modulation was assessed using quantitative real-time polymerase chain reaction analysis (qRT-PCR), Western blotting, and confocal microscopy. Dextran sulfate sodium (DSS) colitis was induced in wild-type, TFF3(-/-), and TLR2(-/-) mice. Recombinant TLR2 ligand or TFF3 peptide were orally administered after DSS termination. Caco-2 cells overexpressing full-length TLR2 or mutant TLR2-R753Q were tested for TFF3 synthesis and functional-related effects in a wounding assay. RESULTS: Data from in vitro (Ls174T) and ex vivo models of murine and human GC reveal that TLR2 activation selectively induces synthesis of TFF3. In vivo studies using TFF3(-/-) or TLR2(-/-) mice demonstrate the ability for oral treatment with a TLR2 agonist to confer antiapoptotic protection of the intestinal mucosa against inflammatory stress-induced damage through TFF3. Recombinant TFF3 rescues TLR2-deficient mice from increased morbidity and mortality during acute colonic injury. Severe ulcerative colitis (UC) has recently been found to be associated with the R753Q polymorphism of the TLR2 gene. The relevance of the observed functional effect of TLR2 in regulating GC is confirmed by the finding that the UC-associated TLR2 R753Q variant is functionally deficient in the ability to induce TFF3 synthesis, thus leading to impaired wound healing. CONCLUSIONS: These data demonstrate a novel function of TLR2 in intestinal GC that links products of commensal bacteria to innate immune protection of the host via TFF3. PMID- 19303022 TI - Clinical challenges and images in GI. Esophageal mucosa expulsion after high-dose chemotherapy. PMID- 19303023 TI - HBsAg seroclearance: the more and earlier, the better. PMID- 19303024 TI - PARP-1 expression in the mouse is controlled by an autoregulatory loop: PARP-1 binding to an upstream S/MAR element and to a novel recognition motif in its promoter suppresses transcription. AB - This work identifies central components of a feedback mechanism for the expression of mouse poly(ADP-ribose) polymerase-1 (PARP-1). Using the stress induced duplex destabilization algorithm, multiple base-unpairing regions (BURs) could be localized in the 5' region of the mouse PARP-1 gene (muPARP-1). Some of these could be identified as scaffold/matrix-attachment regions (S/MARs), suggesting an S/MAR-mediated transcriptional regulation. PARP-1 binding to the most proximal element, S/MAR 1, and to three consensus motifs, AGGCC, in its own promoter (basepairs -956 to +100), could be traced by electrophoretic mobility shift assay. The AGGCC-complementary GGCCT motif was detected by cis-diammine dichloro platinum cross-linking and functionally characterized by the effects of site-directed mutagenesis on its performance in wild type (PARP-1(+/+)) and PARP 1 knockout cells (PARP-1(-/-)). Mutation of the central AGGCC tract at basepairs 554 to -550 prevented PARP-1/promoter interactions, whereby muPARP-1 expression became up-regulated. Transfection of a series of reporter gene constructs with or without S/MAR 1 (basepairs -1523 to -1007) and the more distant S/MAR 2 (basepairs -8373 to -6880), into PARP-1(+/+) as well as PARP-1(-/-) cells, revealed an additional, major level of muPARP-1 promoter down-regulation, triggered by PARP-1 binding to S/MAR 1. We conclude that S/MAR 1 represents an upstream control element that acts in conjunction with the muPARP-1 promoter. These interactions are part of a negative autoregulatory loop. PMID- 19303025 TI - Caffeic acid phenethyl ester accumulates beta-catenin through GSK-3beta and participates in proliferation through mTOR in C2C12 cells. AB - AIM: The aim of this study is to characterize the roles of caffeic acid phenethyl ester (CAPE) in the skeletal muscle cells. MAIN METHODS: We performed immunoblotting assay using various phosphorylation specific antibodies. KEY FINDINGS: We found that CAPE induces rapid and transient phosphorylation of glycogen synthase kinase (GSK)-3beta in a phosphoinositide 3-kinase (PI3K) dependent manner. CAPE also decreases phosphorylation of beta-catenin, ultimately leading to beta-catenin accumulation. In addition, we demonstrated that CAPE activated the mammalian target of rapamycin (mTOR)-p70 S6 ribosomal kinase (S6K) and also stimulated extracellular signal-regulated kinase (ERK). The inhibition of mTOR blocked CAPE-induced ERK phosphorylation. SIGNIFICANCE: Our results suggest that CAPE may act through beta-catenin accumulation via stimulation of GSK-3beta and may also participate in cellular proliferation through the mTOR-ERK pathway. PMID- 19303026 TI - A radiotracer method to study efflux transport of iodide liberated from thyroid hormones via deiodination metabolism in the brain. AB - AIMS: Thyroid hormones (TH) play an important role in the development and functional maintenance of the central nervous system. The purpose of this study was to develop a radiotracer method for studying the in vivo efflux transport of iodide liberated by the TH metabolism in the brain. The rationale of our method is as follows: a radioiodinated compound can enter the brain and rapidly release iodide in situ; the iodide efflux rate can be estimated from the clearance of brain radioactivity after disappearance of the iodinated compound. MAIN METHODS: 6-[(125)I]Iodo-9-pentylpurine ([(125)I]9Pe6IP) was designed to enter the brain and release (125)I(-) by the reaction with glutathione and synthesized from the corresponding bromo derivative in a Br/(125)I exchange reaction. The brain kinetics of radioactivity and radioactive metabolites were investigated after intravenous injection of [(125)I]9Pe6IP into mice. The iodide efflux rate was estimated in mice pretreated with perchlorate, an inhibitor of iodide transport from the brain. KEY FINDINGS: High brain uptake (5.3% injected dose/g) was observed at 1 min, and almost complete conversion of [(125)I]9Pe6IP to (125)I(-) occurred 10 min after injection. The (125)I(-) uptake from the blood was negligible. (125)I(-) was eliminated from the brain along a single-exponential curve with a half-life of 6.0 min. Furthermore, dose-dependent inhibition of (125)I(-) efflux was observed in mice pretreated with perchlorate. SIGNIFICANCE: We conclude that 9Pe6IP labeled with (124)I (positron emitter) or (123)I (single photon emitter) may be useful for studying the in vivo efflux transport of iodide in the brain using nuclear medicine imaging devices. PMID- 19303027 TI - X-ray crystallography of chemical compounds. AB - AIMS: Accurate knowledge of molecular structure is a prerequisite for rational drug design. This review examines the role of X-ray crystallography in providing the required structural information and advances in the field of X-ray crystallography that enhance or expand its role. MAIN METHODS: X-ray crystallography of new drugs candidates and intermediates can provide valuable information of new syntheses and parameters for quantitative structure activity relationships (QSAR). KEY FINDINGS: Crystallographic studies play a vital role in many disciplines including materials science, chemistry, pharmacology, and molecular biology. X-ray crystallography is the most comprehensive technique available to determine molecular structure. A requirement for the high accuracy of crystallographic structures is that a 'good crystal' must be found, and this is often the rate-limiting step. In the past three decades developments in detectors, increases in computer power, and powerful graphics capabilities have contributed to a dramatic increase in the number of materials characterized by X ray crystallography. More recently the advent of high-throughput crystallization techniques has enhanced our ability to produce that one good crystal required for crystallographic analysis. SIGNIFICANCE: Continuing advances in all phases of a crystallographic study have expanded the ranges of samples which can be analyzes by X-ray crystallography to include larger molecules, smaller or weakly diffracting crystals, and twinned crystals. PMID- 19303028 TI - Development of radioligands with optimized imaging properties for quantification of nicotinic acetylcholine receptors by positron emission tomography. AB - AIMS: There is an urgent need for positron emission tomography (PET) imaging of the nicotinic acetylcholine receptors (nAChR) to study the role of the nicotinic system in Alzheimer's and Parkinson's diseases, schizophrenia, drug dependence and many other disorders. Greater understanding of the underlying mechanisms of the nicotinic system could direct the development of medications to treat these disorders. Central nAChRs also contribute to a variety of brain functions, including cognition, behavior and memory. MAIN METHODS: Currently, only two radiotracers, (S)-3-(azetidin-2-ylmethoxy)-2-[(18)F]fluoropyridine (2-[(18)F]FA) and (S)-5-(azetidin-2-ylmethoxy)-2-[(18)F]fluoropyridine (6-[(18)F]FA), are available for studying nAChRs in human brain using PET. However, the "slow" brain kinetics of these radiotracers hamper mathematical modeling and reliable measurement of kinetic parameters since it takes 4-7 h of PET scanning for the tracers to reach steady state. The imaging drawbacks of the presently available nAChR radioligands have initiated the development of radioligands with faster brain kinetics by several research groups. KEY FINDINGS: This minireview attempts to survey the important achievements of several research groups in the discovery of PET nicotinic radioligands reached recently. Specifically, this article reviews papers published from 2006 through 2008 describing the development of fifteen new nAChR (11)C-and (18)F-ligands that show improved imaging properties over 2-[(18)F]FA. SIGNIFICANCE: The continuous efforts of radiomedicinal chemists led to the development of several interesting PET radioligands for imaging of nAChR including [(18)F]AZAN, a potentially superior alternative to 2-[(18)F]FA. PMID- 19303029 TI - Impairment of novelty detection in mice targeted for the Chl1 gene. AB - A deficit in cell adhesion molecules including the human Chl1 (close homologue of the L1 cell adhesion molecule) gene may cause impairment of cognitive processes. Aberrant connectivity in the CA3 region of the hippocampus has been reported in mice lacking the CHL1 protein after Chl1 gene targeting. Previous studies have observed a deficit in the processing of novel information by CHL1-deficient mice. We investigated deficits in spatial discrimination and object discrimination in three groups of mice--Chl1(+/+), Chl1(+/-) and Chl1(-/-)--performing spatial and object novelty tasks. The results indicated that wild-type mice easily recognized objects that were either "displaced" or "substituted". Chl1(-/-) mice showed severe impairment of the capacity to react to both spatial and non-spatial novelty. Chl1(+/-) mice were severely restricted in their ability to detect spatial changes, but succeeded in novel object discrimination. A dose-dependent sensitivity of the organization of the CA3 layer to the CHL1 protein may explain this result. However, the observations suggest that a dysfunction of parts of the brain other than the hippocampus may be involved in the impairment. PMID- 19303030 TI - Flavor-nutrient learning is less rapid with fat than with carbohydrate in rats. AB - Flavor-nutrient learning occurs when the post-ingestive consequences of a food are associated with its flavor. As a signal of the food's energy density, flavor nutrient associations have the potential to contribute to the regulation of meal size. While all calorie sources (fat, carbohydrate, protein, ethanol) can support flavor-nutrient learning, prior research has found that flavor-nutrient associations based on fat may require higher nutrient concentrations and more rigorous experimental protocols than are required to train carbohydrate (cho) based associations. To further explore potential macronutrient-specific differences in flavor-nutrient learning, the present study compared the time course of acquisition of cho- and fat-based associations. Rats were trained to associate distinctive flavors with high-density (3.2 kcal/mL) and low-density (0.2 kcal/mL) orally-consumed solutions, either fat (corn oil emulsion) or carbohydrate (sucrose). For each nutrient, both within- and between-group designs were used to assess (via two-bottle preference testing) whether flavor-nutrient learning had occurred after 2, 4, or 6 training trial pairs. Rats trained with carbohydrate demonstrated preferential intake of the low-density paired flavor after only 2 training pairs; in contrast, rats trained with fat required 6 training pairs. These findings demonstrate differential rapidity of acquisition flavor-nutrient associations. The longer time course of acquisition of fat-based flavor-nutrient associations may be yet another mechanism by which high-fat foods promote overeating. PMID- 19303031 TI - Individual housing of mice--impact on behaviour and stress responses. AB - The replicability of results derived from studies in rodents might be influenced by stress caused by inappropriate housing conditions. Here we compared the experimental behaviour and stress response (circulating corticosterone level and adrenal tyrosine hydroxylase activity) of individually-housed male and female inbred mice with that of animals housed in social groups. All mice were behaviourally tested in the modified hole board test (mHB). Male C57BL/6, BALB/c and A mice housed in groups of 3 were compared with individually-housed mice. In a subsequent experiment female C57BL/6 and BALB/c mice were housed under similar conditions. To exclude the possible effects of within-cage order of testing, only one individual per group was behaviourally tested. Neither male nor female mice housed individually showed stronger signs of stress than their socially-housed counterparts. However, we observed a within-cage order effect on the hormonal stress response (corticosterone) in socially-housed female C57BL/6 mice. No effects of individual housing on behaviour in the mHB were found. PMID- 19303032 TI - The limitation of species range: a consequence of searching along resource gradients. AB - Ecological modelers have long puzzled over the spatial distribution of species. The random walk or diffusive approach to dispersal has yielded important results for biology and mathematics, yet it has been inadequate in explaining all phenomenological features. Ranges can terminate non-smoothly absent a complementary shift in the characteristics of the environment. Also unexplained is the absence of a species from nearby areas of adequate, or even abundant, resources. In this paper, I show how local searching behavior-keyed to a density dependent fitness-can limit the speed and extent of a species' spread. In contrast to standard diffusive processes, pseudo-rational movement facilitates the clustering of populations. It also can be used to estimate the speed of an expanding population range, explain expansion stall, and provides a mechanism by which a population can colonize seemingly removed regions - biogeographic islands in a continental framework. Finally, I discuss the effect of resource degradation and different resource impact/utilization curves on the model. PMID- 19303033 TI - Experimental pathology of local tissue damage induced by Bothrops asper snake venom. AB - Envenomations by Bothrops asper are often associated with complex and severe local pathological manifestations, including edema, blistering, dermonecrosis, myonecrosis and hemorrhage. The pathogenesis of these alterations has been investigated at the experimental level. These effects are mostly the consequence of the direct action of zinc-dependent metalloproteinases (SVMPs) and myotoxic phospholipases A(2) (PLA(2)s). SVMPs induce hemorrhage, blistering, dermonecrosis and general extracellular matrix degradation, whereas PLA(2)s induce myonecrosis and also affect lymphatic vessels. In addition, the prominent vascular alterations leading to hemorrhage and edema may contribute to ischemia and further tissue necrosis. The mechanisms of action of SVMPs and PLA(2)s are discussed in detail in this review. Venom-induced tissue damage plays also a role in promoting bacterial infection. A prominent inflammatory reaction develops as a consequence of these local pathological alterations, with the synthesis and release of abundant mediators, resulting in edema and pain. However, whether inflammatory cells and mediators contribute to further tissue damage is not clear at present. Muscle tissue regeneration after venom-induced pathological effects is often impaired, thus resulting in permanent tissue loss and dysfunction. SVMP induced microvessel damage is likely to be responsible of this poor regenerative outcome. Antivenoms are only partially effective in the neutralization of B. asper-induced local effects, and the search for novel toxin inhibitors represents a potential avenue for improving the treatment of this serious aspect of snakebite envenomation. PMID- 19303034 TI - Experimental pathophysiology of systemic alterations induced by Bothrops asper snake venom. AB - Moderate and severe envenomations by the snake Bothrops asper provoke systemic alterations, such as systemic bleeding, coagulopathy, hypovolemia, hemodynamic instability and shock, and acute renal failure. Systemic hemorrhage is a typical finding of these envenomations, and is primarily caused by the action of P-III snake venom metalloproteinases (SVMPs). This venom also contains a thrombin-like serine proteinase and a prothrombin-activating P-III SVMP, both of which cause defibrin(ogen)ation. Thrombocytopenia, predominantly induced by a C-type lectin like protein, and platelet hypoaggregation, caused by the two defibrin(ogen)ating enzymes, also contribute to hemostatic disturbances, which potentiate the systemic bleeding induced by hemorrhagic SVMPs. Cardiovascular disturbances leading to shock are due to the combined effects of hemorrhagic toxins, other venom components that increase vascular permeability, the action of hypotensive agents in the venom and of endogenous mediators, and the potential cardiotoxic effect of some toxins. Renal alterations are likely to be caused by direct cytotoxicity of venom components in the kidney, and by renal ischemia resultant from hypovolemia and hypoperfusion. Lethality induced by B. asper venom is the consequence of several combined effects among which the action of P-III SVMPs is especially relevant. PMID- 19303035 TI - The supra-additive hyperactivity caused by an amphetamine-chlordiazepoxide mixture exhibits an inverted-U dose response: negative implications for the use of a model in screening for mood stabilizers. AB - One of the few preclinical models used to identify mood stabilizers is an assay in which amphetamine-induced hyperactivity (AMPH) is potentiated by the benzodiazepine chlordiazepoxide (CDP), an effect purportedly blocked by mood stabilizers. Our data here challenge this standard interpretation of the AMPH-CDP model. We show that the potentiating effects of AMPH-CDP are not explained by a pharmacokinetic interaction as both drugs have similar brain and plasma exposures whether administered alone or in combination. Of concern, however, we find that combining CDP (1-12 mg/kg) with AMPH (3 mg/kg) results in an inverted-U dose response in outbred CD-1 as well as inbred C57Bl/6N and 129S6 mice (peak hyperactivity at 3 mg/kg CDP+3 mg/kg AMPH). Such an inverted-U dose response complicates interpreting whether a reduction in hyperactivity produced by a mood stabilizer reflects a "blockade" or a "potentiation" of the mixture. In fact, we show that the prototypical mood stabilizer valproic acid augments the effects of CDP on hypolocomotion and anxiolytic-like behavior (increases punished crossings by Swiss-Webster mice in the four-plate test). We argue that these data, in addition to other practical and theoretical concerns surrounding the model, limit the utility of the AMPH-CDP mixture model in drug discovery. PMID- 19303036 TI - Utility of CSP typing to sub-type clinical Aspergillus fumigatus isolates and proposal for a new CSP type nomenclature. AB - CSP typing is a newly developed sub-typing strategy that employs comparative DNA sequence analysis of the 12-mer tandem repeat region of the AFUA_3G08890 gene. In order to allow standardization of analysis and exchange of results between laboratories, we propose a new nomenclature for individual CSP repeats as well as for CSP types. A collection of 209 clinical isolates of Aspergillus fumigatus recovered from various hospitals throughout The Netherlands was analyzed by using CSP typing and this newly proposed nomenclature. Eighteen different CSP types were recognized, positioning the CSP gene as a typing target between the relatively low discriminatory MLST loci and the highly discriminatory microsatellite markers. CSP typing may be a welcome addition to the existing molecular methods to study the diversity of A. fumigatus at the sub-population level. The results also show the presence of lineages of closely related CSP types within the A. fumigatus population, adding unique and valuable information about the population structure of A. fumigatus. PMID- 19303037 TI - An improved isolation method for attached-living Planctomycetes of the genus Rhodopirellula. AB - Rhodopirellula baltica, an attached-living marine bacterium, was so far isolated as aerobic, heterotrophic bacterium forming pink-to-red colonies on ampicillin containing plates. But many bacteria grow in the presence of ampicillin: marine samples from European Seas contained on average 10,365 colony forming bacteria per ml water sample or sediment. Therefore we developed an improved enrichment protocol to isolate Rhodopirellula strains. To include attached-living bacteria, particles, plankton catches or sediment grains were spread on the plate surface. This was a necessity to reliably obtain new strains. Cycloserine or streptomycin served as additional selective agents. The number of colony forming cells resistant to ampicillin and cycloserine or streptomycin was low, 262 and 107 cfu/ml sample, respectively. Rhodopirellula colonies were identified with a newly developed specific PCR reaction detecting a part of the 16S rRNA gene. The phylogenetic tree of over 60 isolates was established with new primers outside of the 16S rRNA gene and revealed a diversity on the species level in European Seas. PMID- 19303038 TI - A designer biomimetic vector with a chimeric architecture for targeted gene transfer. AB - Designer biomimetic vectors are genetically engineered biomacromolecules that are designed to mimic viral characteristics in order to overcome the cellular barriers associated with the targeted gene transfer. The vector in this study was genetically engineered to contain at precise locations: a) four tandem repeating units of N-terminal domain of histone H2A to condense DNA into stable nanosize particles suitable for cellular uptake, b) a model targeting motif to target HER2 and enhance internalization of nanoparticles, and c) a pH-responsive synthetic fusogenic peptide to disrupt endosome membranes and promote escape of the nanoparticles into the cytosol. The results demonstrate that a fully functional, multi-domain, designer vector can be engineered to target cells with high specificity, overcome the biological barriers associated with targeted gene transfer, and mediate efficient gene transfer. PMID- 19303039 TI - Chitosan based oligoamine polymers: synthesis, characterization, and gene delivery. AB - A series of chitosan-based oligoamine polymers was synthesized from N-maleated chitosan (NMC) via Michael addition with diethylenetriamine (DETA), triethylenetetramine (TETA), tetraethylenepentamine (TEPA) and linear polyethylenimine (M(n) 423), respectively. The resulted polymers exhibited well binding ability to condense plasmid DNA to form complexes with size ranging from 200 to 600 nm when the polymer/DNA weight ratio was above 7. The polymer/DNA complexes observed by scanning electron microscopy (SEM) exhibited a compact and spherical morphology. The cytotoxicity assay showed that the synthesized polymers were less toxic than that of PEI(25 K). The gene transfection effect of resulted polymers was evaluated in 293T and HeLa cells, and the results showed that the gene transfection efficiency of these polymers was better than that of chitosan. Moreover, the transfection efficiency was dependent on the length of the oligoamine side chains and the molecular weight of the chitosan derivatives. PMID- 19303040 TI - A 35-day gavage safety assessment of ginger in rats. AB - Ginger (Zingiber officinale Roscoe, Zingiberacae) is one of the most commonly used spices around the world and a traditional medicinal plant that has been widely used in Chinese, Ayurvedic and Unani-Tibb medicines for several thousand years. However, there was still lack of systemic safety evaluation. We conducted a 35-day toxicity study on ginger in rats. Both male and female rats were daily treated with ginger powder at the dosages of 500, 1000 and 2000 mg/kg body weight by a gavage method for 35 days. The results demonstrated that this chronic administration of ginger was not associated with any mortalities and abnormalities in general conditions, behavior, growth, and food and water consumption. Except for dose-related decrease in serum lactate dehydrogenase activity in males, ginger treatment induced similar hematological and blood biochemical parameters to those of controlled animals. In general, ginger treatment caused no overt organ abnormality. Only at a very high dose (2000 mg/kg), ginger led to slightly reduced absolute and relative weights of testes (by 14.4% and 11.5%, respectively). This study provides a new understanding of the toxicological properties of ginger. PMID- 19303041 TI - Drug-induced liver injury following positive drug rechallenge. AB - Drug rechallenge (or reinitiation), following an event of drug-induced liver injury, can lead to serious or fatal liver injury. A retrospective review of a large pharmaceutical safety database was conducted to assess clinical outcomes of positive drug rechallenge following possible drug-induced liver injury. Positive rechallenge with suspect drug was reported in 770 of 36,795 hepatic adverse events. A total of 88 cases met inclusion criteria for analysis. Mean age was 44 years (range 0.5-83) and 56% were male. A broad spectrum of suspect drugs were identified. Many patients exhibited hepatitis symptoms or jaundice on the initial and rechallenge liver event. Twelve patients (14%) exhibited clinically worrisome severe hepatocellular injury and jaundice on either initial or rechallenge event and two died, reflecting a 2.3% fatality rate in those with positive rechallenge. The two fatalities developed severe hepatocellular injury with jaundice only upon rechallenge. Liver injury recurred in most rechallenges. Improved identification and communication of possible drug-induced liver injury is needed to avoid potentially serious and/or fatal drug rechallenges. Clinicians should generally avoid such rechallenges. PMID- 19303042 TI - A case of psychosis associated with left thalamic lacunar infarcts. PMID- 19303043 TI - Antioxidant and antidermatophytic activities of essential oil and extracts of Metasequoia glyptostroboides Miki ex Hu. AB - This study was undertaken to assess the antioxidant and antidermatophytic potential of the essential oil and extracts (hexane, chloroform, ethyl acetate and methanol) of Metasequoia glyptostroboides Miki ex Hu. Antioxidant activity was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The free radical scavenging activities of the oil and ethyl acetate extract were found to be superior (IC(50)=9.1 and 14.24 microg/ml, respectively) as compared to butylatedhydroxyanisole (BHA), (IC(50)=18.27 microg/ml). Also the ethyl acetate extract revealed the highest phenolic contents (93.26 mg/g of dry wt) as compared to the other extracts. Further, oil (1250 microg/disc) and extracts (1750 microg/disc) revealed 35.33-67.66 and 18.0-53.3% antidermatophytic effect, respectively, along with their respective MIC values (62.5-500 and 250-4000 microg/ml) against Trichophyton rubrum KCTC 6345, T. rubrum KCTC 6375, T. rubrum KCTC 6352, T. mentagrophytes KCTC 6085, T. mentagrophytes KCTC 6077, T. mentagrophytes KCTC 6316, Microsporum canis KCTC 6591, M. canis KCTC 6348 and M. canis KCTC 6349. The oil also had a strong detrimental effect on spore germination as well as concentration and time-dependent kinetic inhibition of M. canis KCTC 6591. PMID- 19303044 TI - The essential role of lipids in Alzheimer's disease. AB - In the absence of efficient diagnostic and therapeutic tools, Alzheimer's disease (AD) is a major public health concern due to longer life expectancy in the Western countries. Although the precise cause of AD is still unknown, soluble beta-amyloid (Abeta) oligomers are considered the proximate effectors of the synaptic injury and neuronal death occurring in the early stages of AD. Abeta oligomers may directly interact with the synaptic membrane, leading to impairment of synaptic functions and subsequent signalling pathways triggering neurodegeneration. Therefore, membrane structure and lipid status should be considered determinant factors in Abeta-oligomer-induced synaptic and cell injuries, and therefore AD progression. Numerous epidemiological studies have highlighted close relationships between AD incidence and dietary patterns. Among the nutritional factors involved, lipids significantly influence AD pathogenesis. It is likely that maintenance of adequate membrane lipid content could prevent the production of Abeta peptide as well as its deleterious effects upon its interaction with synaptic membrane, thereby protecting neurons from Abeta-induced neurodegeneration. As major constituents of neuronal lipids, n-3 polyunsaturated fatty acids are of particular interest in the prevention of AD valuable diet ingredients whose neuroprotective properties could be essential for designing preventive nutrition-based strategies. In this review, we discuss the functional relevance of neuronal membrane features with respect to susceptibility to Abeta oligomers and AD pathogenesis, as well as the prospective capacities of lipids to prevent or to delay the disease. PMID- 19303045 TI - Splice variants of the human ZC3H14 gene generate multiple isoforms of a zinc finger polyadenosine RNA binding protein. AB - The human ZC3H14 gene encodes an evolutionarily conserved Cys(3)His zinc finger protein that binds specifically to polyadenosine RNA and is thus postulated to modulate post-transcriptional gene expression. Expressed sequence tag (EST) data predicts multiple splice variants of both human and mouse ZC3H14. Analysis of ZC3H14 expression in both human cell lines and mouse tissues confirms the presence of multiple alternatively spliced transcripts. Although all of these transcripts encode protein isoforms that contain the conserved C-terminal zinc finger domain, suggesting that they could all bind to polyadenosine RNA, they differ in other functionally important domains. Most of the alternative transcripts encode closely related proteins (termed isoforms 1, 2, 3, and 3 short) that differ primarily in the inclusion of three small exons, 9, 10, and 11, resulting in predicted protein isoforms ranging from 82 to 64 kDa. Each of these closely related isoforms contains predicted classical nuclear localization signals (cNLS) within exons 7 and 11. Consistent with the presence of these putative nuclear targeting signals, these ZC3H14 isoforms are all localized to the nucleus. In contrast, an additional transcript encodes a smaller protein (34 kDa) with an alternative first exon (isoform 4). Consistent with the absence of the predicted cNLS motifs located in exons 7 and 11, ZC3H14 isoform 4 is localized to the cytoplasm. Both EST data and experimental data suggest that this variant is enriched in testes and brain. Using an antibody that detects endogenous ZC3H14 isoforms 1-3 reveals localization of these isoforms to nuclear speckles. These speckles co-localize with the splicing factor, SC35, suggesting a role for nuclear ZC3H14 in mRNA processing. Taken together, these results demonstrate that multiple transcripts encoding several ZC3H14 isoforms exist in vivo. Both nuclear and cytoplasmic ZC3H14 isoforms could have distinct effects on gene expression mediated by the common Cys(3)His zinc finger polyadenosine RNA binding domain. PMID- 19303046 TI - A cotton gene encodes a tonoplast aquaporin that is involved in cell tolerance to cold stress. AB - To enhance the survival probability in cold stress, plant cells often increase their cold- and freezing-tolerance in response to low, nonfreezing temperatures by expressing some cold-related genes. In present study, a cotton gene encoding tonoplast intrinsic protein (TIP) was isolated from a cotton seedling cDNA library, and designated as GhTIP1;1. GFP fluorescent microscopy indicated that GhTIP1;1 protein was localized to the vacuolar membrane. Assay on GhTIP1;1 expression in Xenopus laevis oocytes demonstrated that GhTIP1;1 protein displayed water channel activity and facilitated water transport to the cells. At normal conditions, GhTIP1;1 transcripts were predominantly accumulated in roots and hypocotyls, but less abundance in other tissues of cotton. The GhTIP1;1 expression was dramatically up-regulated in cotyledons, but down-regulated in roots within a few hours after cotton seedlings were cold-treated. Overexpression of GhTIP1;1 in yeast (Schizosaccharomyces pombe) significantly enhanced the cell survival probability, suggesting that the GhTIP1;1 protein is involved in cell freezing-tolerance. PMID- 19303047 TI - Isolation of active regulatory elements from eukaryotic chromatin using FAIRE (Formaldehyde Assisted Isolation of Regulatory Elements). AB - The binding of sequence-specific regulatory factors and the recruitment of chromatin remodeling activities cause nucleosomes to be evicted from chromatin in eukaryotic cells. Traditionally, these active sites have been identified experimentally through their sensitivity to nucleases. Here we describe the details of a simple procedure for the genome-wide isolation of nucleosome depleted DNA from human chromatin, termed FAIRE (Formaldehyde Assisted Isolation of Regulatory Elements). We also provide protocols for different methods of detecting FAIRE-enriched DNA, including use of PCR, DNA microarrays, and next generation sequencing. FAIRE works on all eukaryotic chromatin tested to date. To perform FAIRE, chromatin is crosslinked with formaldehyde, sheared by sonication, and phenol-chloroform extracted. Most genomic DNA is crosslinked to nucleosomes and is sequestered to the interphase, whereas DNA recovered in the aqueous phase corresponds to nucleosome-depleted regions of the genome. The isolated regions are largely coincident with the location of DNaseI hypersensitive sites, transcriptional start sites, enhancers, insulators, and active promoters. Given its speed and simplicity, FAIRE has utility in establishing chromatin profiles of diverse cell types in health and disease, isolating DNA regulatory elements en masse for further characterization, and as a screening assay for the effects of small molecules on chromatin organization. PMID- 19303048 TI - Immunomodulatory consequences of ODN CpG-polycation complexes. AB - Immunostimulatory ODN CpGs have extensively been tested as adjuvants and immunotherapeutics and hold a lot of promise for human use. In our studies we took advantage of their negative charge to study their biological activities after being complexed with carbon nanotubes, a novel vector for vaccine delivery and Tat protein of HIV, a target protein for therapeutic or prophylactic intervention. In the case of carbon nanotubes, ODN CpGs were able to form stable complexes based on charge interaction and exert increased immunostimulatory activity in vitro. With regard to the Tat protein, ODN CpGs were shown to bind effectively through the basic domain of the protein representing residues 44-61. Moreover, using surface Plasmon Resonance Technology and an in vitro cellular system, ODN CpGs were shown to inhibit the interaction of Tat protein with the transactivation responsive element, a bulged RNA hairpin structure. However, when ODN CpGs were complexed with Tat they readily increased the apoptotic properties of this protein as studied in CD3-stimulated Jurkat cells. Overall, our findings together with published data support the view that for harnessing the beneficial effects of ODN CpGs a careful consideration has to be given depending on the target intervention. PMID- 19303049 TI - Detection and interpretation of expression quantitative trait loci (eQTL). AB - Analysis of expression quantitative trait loci (eQTL) provides a means for detecting transcriptional regulatory relationships at a genome-wide scale. Here we explain the eQTL analysis pipeline, we introduce publicly available tools for the statistical analysis, and we discuss issues that might complicate the eQTL mapping process. The detection and interpretation of eQTL requires careful consideration of a range of potentially confounding effects. Particularly population substructure and batch effects may lead to the detection of many false positive eQTL if not accounted for. Traditionally, most eQTL mapping methods only check for the correlation of single loci with gene expression. In order to detect (epistatic) interactions between distant genetic loci one has to take into account several loci simultaneously. Here, we present the Random Forest regression method as a way of accounting for interacting loci. Next, we introduce analysis methods aiding the biological interpretation of detected eQTL. For example, the notion of local (cis) and distant (trans) eQTL has been very useful for interpreting the causes and implications of eQTL in many studies. In addition, Bayesian networks have been used extensively to infer causal relationships among eQTL and between eQTL and other genetic associations (e.g. disease associated loci). Also, the integration of eQTL with complementary information such as physical protein interaction data may significantly improve statistical power and provide insight into possible molecular mechanisms linking the regulator to its target gene. The eQTL approach is potentially very powerful for the analysis of regulatory pathways affecting disease susceptibility and other relevant traits. However, careful analysis is required to unleash its full potential. PMID- 19303050 TI - Abolition of reflex bradycardia by cardiac vagotomy has no effect on the regulation of oxygen uptake by Atlantic cod in progressive hypoxia. AB - The functional significance of chemoreflexive hypoxic bradycardia was explored in Atlantic cod Gadus morhua L. (mean mass approximately 800 g, acclimated to a seawater temperature of 11 degrees C) by investigating responses to progressive hypoxia following section of the cardiac branches of cranial nerve X. Cardiac denervation had no effect on oxygen uptake rate (MO(2)), gill ventilation rate (f(G)) or opercular pressure amplitude (P(OP)) under normoxic conditions, but caused a significant increase in heart rate (f(H)), to 50+/-1 beats min(-1) by comparison to 40+/-2 beats min(-1) in sham-operated cod (mean+/-s.e.m., n=9). Sham-operated cod exhibited transient profound bradycardia following oxygen chemoreceptor stimulation by bolus injection of sodium cyanide into the buccal cavity (2 mg in 2 ml seawater), but this cardiac chemoreflex was abolished in denervated cod. Both groups, however, exhibited similar marked transient chemoreflexive hyperventilation following NaCN. When exposed from normoxia (PO(2) approximately 18 kPa) to progressive hypoxia at nominal water PO(2)'s of 8, 6, 5, 4 and 3 kPa, both groups exhibited the same pattern of homeostatic regulation of MO(2), with no significant difference in their mean critical PO(2) (P(crit)) values, which were 7.40+/-0.81 kPa and 8.73+/-0.71 kPa, respectively (n=9). Both groups exhibited significant bradycardia during progressive hypoxia, although denervated fish always had higher mean f(H). The incipient threshold for bradycardia coincided with P(crit) in sham-operated cod whereas, in denervates, the threshold was below their P(crit) and bradycardia presumably reflected direct effects of hypoxia on the myocardium. The sham-operated group displayed a significantly more pronounced ventilatory response than denervates in hypoxia, in particular for P(OP). In sham-operated cod, peak ventilatory responses occurred in deep hypoxia below P(crit) whereas, in denervates, more modest peak responses coincided with P(crit) and, in deep hypoxia, they exhibited a significant decline in f(G) below their normoxic rate. Only a minority of shams lost equilibrium in hypoxia whereas a majority of denervates did, some of which failed to recover. The results indicate that chemoreflexive bradycardia plays no role in the homeostatic regulation of oxygen uptake by cod in hypoxia, but does contribute to maintenance of overall functional integrity below P(crit). PMID- 19303051 TI - First evidence for the salt-dependent folding and activity of an esterase from the halophilic archaea Haloarcula marismortui. AB - A gene encoding an esterase from Haloarcula marismortui, a halophilic archaea from the Dead Sea, was cloned, expressed in Escherichia coli, and the recombinant protein (Hm EST) was biochemically characterized. The enzymatic activity of Hm EST was shown to exhibit salt dependence through salt-dependent folding. Hm EST exhibits a preference for short chain fatty acids and monoesters. It is inhibited by phenylmethylsulfonyl fluoride, diethyl-p-nitrophenyl phosphate, and 5-methoxy 3-(4-phenoxyphenyl)-3H-[1,3,4]oxadiazol-2-one, confirming the conclusion from sequence alignments that Hm EST is a serine carboxylesterase belonging to the hormone-sensitive lipase family. The activity of Hm EST is optimum in the presence of 3 M KCl and no activity was detected in the absence of salts. Far-UV circular dichroism showed that Hm EST is totally unfolded in salt-free medium and secondary structure appears in the presence of 0.25-0.5 M KCl. After salt depletion, the protein was able to recover 60% of its initial activity when 2 M KCl was added. A 3D model of Hm EST was built and its surface properties were analyzed, pointing to an enrichment in acidic residues paralleled by a depletion in basic residues. This peculiar charge repartition at the protein surface supports a better stability of the protein in a high salt environment. PMID- 19303052 TI - Structure of the Malpighian tubule cells and annual changes in the structure and chemical composition of their spherites in the cave cricket Troglophilus neglectus Krauss, 1878 (Rhaphidophoridae, Saltatoria). AB - Periodical changes in the structure of spherites in the Malpighian tubule cells of the cave cricket Troglophilus neglectus were studied to elucidate their role during the cricket's life cycle in natural circumstances. Special interest was given to the dormant overwintering period when we hypothesized that the primary role of spherites is to supply minerals for basic vital processes. The investigation was carried out by light and transmission electron microscopy, energy dispersive X-ray spectroscopy, electron energy-loss spectroscopy and energy-filtering TEM. Spherites are present only in the middle Malpighian tubule segment, consisting of Type 1 cells, characterized, among other features, by a round, apically placed nucleus and numerous spherites, and a few Type 2 cells with an elongated nucleus in the centre and sparse spherites. At the beginning of dormancy in November juveniles, minerals are accumulated in spherites and then decline until March. In one-year-old May larvae, spherites are commonly rich in minerals, and from July onwards they are progressively exploited in the adults. Spherite destruction starts with apoptosis in senile October individuals. The findings suggest that the mineral supply of spherites in Malpighian tubules is crucial to supporting vital processes throughout the life cycle of T. neglectus. PMID- 19303055 TI - Best clinical and research practice in epilepsy of older people: Focus on antiepileptic drug adherence. AB - Few studies have examined the issues that are specific to the older person with epilepsy, a population of increasing prominence in epilepsy management. Our understanding of the impact of epilepsy in the older person is based predominantly on what is inferred from studies of younger adults. Consequently, there is relatively little documented about the impact of epilepsy on the everyday lives of older people. In this article, we focus on adherence and its consequences for the physical, social, and psychological well-being of the older person. A number of strategies are proposed to improve adherence, including patient education through better communication between physician and patient; simplification of the medical regime; and use of extended-release formulations. This issue highlights that to ameliorate the impact of epilepsy for the older person with epilepsy, a greater understanding is required so that appropriate interventions can be tailored. PMID- 19303056 TI - Cytotoxicity and hemolytic activity of jellyfish Nemopilema nomurai (Scyphozoa: Rhizostomeae) venom. AB - The recent bloom of a giant jellyfish Nemopilema nomurai has caused a danger to sea bathers and fishery damages in the waters of China, Korea, and Japan. The present study investigated the cytotoxic and hemolytic activities of crude venom extract of N. nomurai using a number of in vitro assays. The jellyfish venom showed a much higher cytotoxic activity in H9C2 heart myoblast than in C2C12 skeletal myoblast (LC(50)=2 microg/mL vs. 12 microg/mL, respectively), suggesting its possible in vivo selective toxicity on cardiac tissue. This result is consistent with our previous finding that cardiovascular function is a target of the venom. In order to determine the stability of N. nomurai venom, its cytotoxicity was examined under the various temperature and pH conditions. The activity was relatively well retained at low environmental temperature (or=60 degrees C). In pH stability test, the venom has abruptly lost its activity at low pH environment (pH or =6.10 mmol/l and 2hPG<7.80 mmol/l), isolated post-load hyperglycaemia (IPH, FPG<6.10 mmol/l and 2hPG> or =7.80 mmol/l) and combined fasting and post-load hyperglycaemia (CH, FPG> or =6.10 mmol/l and 2hPG> or =7.80 mmol/l). Multivariate-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) for CVD mortality were estimated using Cox proportional hazard analysis. RESULTS: Multivariate-adjusted HRs (95% CIs) for high-density lipoprotein cholesterol (HDL-C) were 0.84 (0.75-0.94), 0.66 (0.48 0.92), 1.03 (0.84-1.27) and 0.67 (0.51-0.89) in individuals with normoglycaemia, IFH, IPH and CH, respectively. For total cholesterol (TC) to HDL-C ratio they were 1.14 (1.03-1.27), 1.44 (1.13-1.84), 0.94 (0.77-1.15) and 1.26 (1.05-1.50), respectively. HRs for TC and triglycerides (TG) were not significant in most of the glucose categories except for TG in those with CH [HR 1.12 (1.00-1.27)]. CONCLUSIONS: Low HDL-C and high TC/HDL-C increase CVD mortality in either diabetic or non-diabetic individuals defined based on the fasting glucose criteria, but not the 2-h criteria. TG is a significant CVD risk predictor only in the presence of combined hyperglycaemia or diabetes. The difference between fasting and post-load hyperglycaemia with regard to the lipid-CVD relation may suggest a different pathophysiology underlying these two prediabetic states. PMID- 19303073 TI - Muscle-induced accelerations at maximum activation to assess individual muscle capacity during movement. AB - Analyses of muscle-induced accelerations provide insight into how individual muscles contribute to motion. In previous studies, investigators have calculated muscle-induced accelerations on a per unit force basis to assess the potential of individual muscles to contribute to motion. However, because muscle force is a function of muscle activation, length, and shortening velocity, examining induced accelerations per unit force does not take into account how the capacity of individual muscles to produce force changes during movement. Alternatively, calculating a muscle's induced accelerations at maximum activation considers the extent to which the muscle can produce force during movement, as well as the potential of the muscle to accelerate the joints at each instant due to its moment arm(s) and the dynamics of the system. We computed both quantities for the major lower extremity muscles active during the stance phase of normal gait. We found that analyzing the induced accelerations at maximum activation in some cases led to a different interpretation of the muscles' potential actions than analyzing the induced accelerations per unit force. For example, per unit force, gluteus maximus has a very large potential to accelerate the knee during single limb stance, but only a small potential to accelerate the knee at maximum activation due to this muscle operating in suboptimal regions of its force-length velocity curve during the majority of stance. This new analysis technique will be useful in studying abnormal movement, when altered kinematics may influence the capacity of muscles to accelerate joints due to altered lengths and shortening velocities. PMID- 19303074 TI - Assessment of specific migration to aqueous simulants of a new active food packaging containing essential oils by means of an automatic multiple dynamic hollow fibre liquid phase microextraction system. AB - The determination of specific migration in the three aqueous food simulants (water, 3% acetic acid and 10% ethanol) from experimental active packaging polypropylene-based films containing natural essential oils as active agents has been carried out for the first time by a two-phase hollow fibre liquid phase microextraction (HFLPME). Due to the high number of variables involved, an experimental design has been applied. High throughput, with six samples running simultaneously in a highly automated system working in dynamic extraction mode, has been achieved. The main analytical characteristics are detection limits as low as 0.01 microg kg(-1), linearity higher than 0.99 for almost 5 magnitude orders, average precision below 16% as RSD and concentration factors ranging from 4 to 189. Migration of 43 compounds including terpenes, alkanes, plastic additives and degradation compounds is reported. According to the results obtained and European legislation, the packaging prototypes tested could be safely marketed. PMID- 19303075 TI - Gas chromatography/mass spectrometry analysis of components of pyridine temperature-programmed desorption spectra from surface of copper-supported catalysts. AB - The method of pyridine temperature-programmed desorption (TPD) was applied for the measurement of acid properties of in situ reduced copper catalysts on silicate support. A thermal-conductivity detector (TCD) was used for the detection of TPD spectra of pyridine. The combination of flame-ionization detector and thermal conductivity detector shows that the region of TPD spectrum with the peak maxima T(MAX1)=350 degrees C is a superposition of the TCD response on spectra of desorbed pyridine, water and carbon dioxide, desorbing simultaneously from the catalyst surface. The method for the elimination of H(2)O and CO(2) on the layer of NaOH was tested and the pure TPD spectrum of pyridine was obtained. The exact determination of pyridine concentration allows to estimate the amount of weak and medium acid centers of the catalyst. The gas chromatography with the mass spectroscopy (GC-MS) analyses was used for the interpretation of high temperature region of the pyridine TPD spectra (T(MAX2)=620 degrees C). It was found that pyridine bonded on the strong acid centers is decomposed to N(2) and CO under very high temperature. The available chromatographic method for the separation of components present in pyridine TPD spectrum in the high-temperature region was suggested. The method for the quantification of strong acidity of copper-supported catalyst was found. PMID- 19303076 TI - Improvement of proteome coverage using hydrophobic monolithic columns in shotgun proteome analysis. AB - Monolithic columns are widely used in shotgun proteome analysis. However, it is difficult to increase the separation capability and proteome coverage by using conventionally organic polymer-based monolithic column due to the difficulty of controlling homogeneity of the overall pore structure (both pores and microglobules), which leads to relatively low column efficiency. Therefore, we studied the effect of constitute and percentage of porogenic solvent, functional monomer, column length, and separation gradient on the peak capacity and proteome coverage by methacrylate-based reversed phase monolithic columns. It was demonstrated that the porous property of the hydrophobic monolith, which was mainly determined by the porogenic solvent, was crucial to the proteome coverage when similar methacrylate monomer was utilized and a ternary porogenic solvent was adopted to prepare C12 monolithic column with relatively homogeneous overall pore structure. It was also shown that high proteome coverage could be reliably obtained with online multidimensional separation using totally monolithic columns system with the length of analytical column at 85 cm and reversed phase separation gradient at 210 min. PMID- 19303077 TI - Negative-pressure cavitation extraction for the determination of flavonoids in pigeon pea leaves by liquid chromatography-tandem mass spectrometry. AB - A new method, namely negative-pressure cavitation extraction (NPCE), followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) is presented for the extraction and quantification of flavonoids in pigeon pea leaves. This method combines the high efficiency of NPCE and the sensitivity and accuracy of MS/MS. The influential parameters of the NPCE procedure including liquid/solid ratio, extraction time, nitrogen flow and number of extraction cycles were optimized. Under optimized conditions, the efficiency of NPCE for extracting five flavonoids was compared to microwave-assisted extraction (MAE), ultrasonic extraction (USE) and heating reflux extraction (HRE). Additionally, structural disruption to pigeon pea leaves samples with different extraction methods was investigated by scanning electron microscopy. The relative recovery with NPCE was equivalent to or higher than that with USE and obviously higher than those with MAE and HRE which are usually conducted in higher temperatures. Furthermore, because NPCE was performed with nitrogen at room temperature, the degradation and oxidation of analytes were avoided. In addition, the NPCE method was validated in terms of repeatability and reproducibility, relative standard deviation for relative recovery was lower than 5.84 and 8.83%, respectively. The method was also successfully applied for the quantification of five flavonoids in pigeon pea leaves. All these results suggest that the developed NPCE-LC-MS/MS method represents an excellent alternative for the extraction and quantification of flavonoids in other plant materials. PMID- 19303078 TI - Metal complexes stability constant determination by hyphenation of capillary electrophoresis with inductively coupled plasma mass spectrometry: the case of 1:1 metal-to-ligand stoichiometry. AB - Nuclear energy development has raised new issues like radionuclides biogeochemistry. The modelling of their biochemical properties involves the accurate determination of thermodynamical data, like stability constants. This can be achieved by using hyphenated capillary electrophoresis (CE)-ICPMS and the method was applied successfully on 1:1 lanthanum-oxalate and uranyl-oxalate complexes. Several significant steps are discussed: choice of analytical conditions, electrophoretic mobility calculation, mathematical treatment of experimental data by using linear regressions, ligand concentration and ionic strength corrections. The following values were obtained with a good precision for lanthanum-oxalate and uranyl-oxalate complexes: log(K degrees (LaOxa(+)))=6.10+/-0.10 and log(K degrees (UO(2)Oxa))=6.40+/-0.30, respectively, at infinite dilution. These values are consistent with the literature data, showing CE-ICPMS potential for metal complexes stability constants determination. PMID- 19303079 TI - Histopathological examination of the pancreas of the Koala (Phascolarctos cinereus). AB - This retrospective study, which was based on koala pancreatic specimens taken 2, 24, 48 and 72h after death, showed that the degree of autolysis did not necessarily exclude histopathological examination. Disorders not previously reported in the pancreas of koalas included the following: inflammation and necrosis; atrophy and fibrosis of exocrine pancreatic tissue; lymphosarcoma; pancreatic heterotopy; and ductal adenocarcinoma. PMID- 19303080 TI - Nicardipine use in cerebrovascular disease: a review of controlled clinical studies. AB - Nicardipine is a dihydropyridine-type Ca(2+) channel blocker (CCB) with strong antihypertensive activity and with a peculiar cerebrovascular profile. This paper has reviewed the main controlled clinical studies on nicardipine in pathologies associated with cerebrovascular impairment. Subarachnoid haemorrhage (SAH) is managed with CCBs to prevent vasospasm and improve clinical outcomes. Nimodipine is the CCB licensed for this indication. Former studies did not demonstrate an advantage of nicardipine versus nimodipine in SAH. A more recent approach administering the drug intra-arterially or using implants of nicardipine prolonged-release showed a decreased incidence of vasospasm, delayed ischemic deficits and improved clinical outcome after severe SAH. Nicardipine is recommended for elevated blood pressure after acute ischemic stroke or intracerebral haemorrhage and is effective in prevention of stroke. More recent investigations were focused on the treatment of cognitive deterioration of vascular origin. In this setting nicardipine has been investigated in more than 6000 patients, with improvement of cognitive deterioration in more than 60% of patients treated. The anti-hypertensive activity of nicardipine, its safety and effectiveness in cognitive domain, suggests re-considering this drug in the treatment of cognitive impairment of vascular origin and for reducing the risk of recurrent stroke in patients at high risk of it. PMID- 19303081 TI - The relationship between subjective reports of fatigue and executive control in multiple sclerosis. AB - Previous studies failed to show a relationship between fatigue and cognitive performance. We used a theory-based Delayed Item Recognition (DIR) paradigm to examine the hypothesis that subjective reports of fatigue and executive control processes were related in MS. Participants were 20 individuals diagnosed with definite diagnosis of MS with Relapsing-Remitting course and 20 controls case matched for age, sex, education and IQ. The DIR paradigm manipulated executive demands in three conditions: Alone, Partial Interference (PI), and Complete Interference (CI). Fatigue was assessed using the Fatigue Severity Scale (FSS). RESULTS: ANOVA Repeated measures analyses showed that DIR performance was slower and less accurate as a function of MS and increased executive demands across the three task conditions. Separate linear regressions revealed that fatigue was related to DIR reaction time and accuracy performance only in the CI condition where executive demands are maximized, and only in the MS group. The present study provided first behavioral evidence that fatigue and executive control are uniquely related in MS. PMID- 19303082 TI - Cross-cultural adaptation and validation of the Portuguese version of the Knee injury and Osteoarthritis Outcome Score (KOOS). AB - OBJECTIVE: The objective of this study was to translate and culturally adapt the Knee injury and Osteoarthritis Outcome Score (KOOS) to the Portuguese language and to test its reliability, validity, floor/ceiling effects and responsiveness. METHODS: This new version was obtained with forward/backward translations, consensus panels and a pre-test. The Portuguese KOOS and Medical Outcomes Study - 36 item Short Form (SF-36) questionnaires, visual analogue scales (VAS) of pain, disability and discomfort, and a form for the characteristics of the patients were applied to 223 subjects with knee osteoarthritis (OA). RESULTS: Reliability was acceptable with Cronbach's alpha coefficients between 0.77 and 0.95, and intraclass correlation coefficients (ICC) ranging from 0.82 to 0.94 for the KOOS subscales. Construct validity was supported by the confirmation of six of the seven predefined hypotheses involving expected correlations between KOOS subscales, SF-36 subscales and VAS. An additional predefined hypothesis was also confirmed with the subjects that need walking aids obtaining lower scores in all five KOOS subscales (P< or =0.001). Floor/ceiling effects were considered to be not present, except for the subscale function in sport and recreation (33.6% of the subjects reported worst possible score). Responsiveness to 4 weeks of physical therapy was demonstrated with standardized effect size between 0.78 and 1.08, and standardized response mean ranging from 0.83 to 1.37 for the KOOS subscales. CONCLUSION: The Portuguese KOOS evidenced acceptable psychometric characteristics. PMID- 19303083 TI - Re: Artificial urinary sphincter in children-voiding or emptying? An evaluation of functional results in 44 patients M. Catti, S. Lortat-Jacob, M. Morineau and H. Lottmann J Urol 2008; 180: 690-693. PMID- 19303084 TI - Re: The performance of prostate specific antigen for predicting prostate cancer is maintained after a prior negative prostate biopsy I. M. Thompson, C. M. Tangen, D. P. Ankerst, C. Chi, M. S. Lucia, P. Goodman, H. Parnes and C. A. Coltman, Jr. J Urol 2008; 180: 544-547. PMID- 19303085 TI - Re: The malone antegrade continence enema: single institutional review A. H. Bani Hani, M. P. Cain, M. Kaefer, K. K. Meldrum, S. King, C. S. Johnson and R. C. Rink J Urol 2008; 180: 1106-1110. PMID- 19303086 TI - Re: Best practice statement on cryosurgery for the treatment of localized prostate cancer R. J. Babaian, B. Donnelly, D. Bahn, J. G. Baust, M. Dineen, D. Ellis, A. Katz, L. Pisters, D. Rukstalis, K. Shinohara and J. B. Thrasher J Urol 2008; 180: 1993-2004. PMID- 19303087 TI - Body mass index as a predictive value for complications associated with reconstructive surgery in patients with myelodysplasia. AB - PURPOSE: Patients with myelodysplasia often have urological pathology, with 25% to 40% requiring reconstructive procedures to achieve urinary and/or fecal continence. Complication rates from these major reconstructive procedures range between 10% and 50%. Additionally many of these patients have significant comorbidities, including a nonambulatory status that leads to an increased body mass index. It is currently unknown whether a high body mass index is associated with increased surgical complications. In this study we compare body mass index and postoperative complications. MATERIALS AND METHODS: We retrospectively reviewed the charts of all patients with myelodysplasia undergoing urinary or fecal reconstructive procedures. We analyzed data for body mass index and any documented complication occurring during hospitalization or at any time during followup. Patients were categorized based on body mass index as normal weight (less than 85th percentile), overweight (85th to 95th percentile) or obese (greater than 95th percentile). Statistical analyses using chi-square and Fisher's exact tests were then performed. RESULTS: Reconstructive procedures were carried out in 66 patients with myelodysplasia between 1997 and 2005. A total of 48 bladder augmentations were performed with a total of 101 stomas created. Mean followup was 39 months. Height and weight were available for body mass index calculation in 60 patients. Obesity was common in our patients with myelodysplasia, affecting 33% of the population (20 of 60 patients). We found a total of 53 complications in 31 patients (52%). There was a significant association between presence of complications and weight category, with complications occurring in 40% of normal weight, 40% of overweight and 75% of obese patients (p = 0.0380). An association between stomal stenosis and weight category was also found (p = 0.0373). In addition, multiple complications were more prevalent in obese patients. Of the 15 patients (25%) with 2 or more complications 10 (67%) were obese (p = 0.0066). CONCLUSIONS: Patients with myelodysplasia have a high incidence of obesity. Since obesity is associated with a higher complication rate, weight loss programs are highly recommended for obese patients with myelodysplasia before and after any reconstructive surgery. PMID- 19303089 TI - Hemostatic role of a folkloric medicinal plant extract in a rat partial nephrectomy model: controlled experimental trial. AB - PURPOSE: Ankaferd BloodStopper is a mixture of 5 plants used in traditional Turkish medicine as hemostatic agent for external traumatic, postoperative and dental bleeding. We investigated the hemostatic efficacy of Ankaferd BloodStopper for partial nephrectomy. MATERIALS AND METHODS: A total of 24 Wistar rats were divided into 4 groups of 6 each. Group 1 underwent partial nephrectomy with hilar control as the conventional technique. Group 2 underwent the conventional technique with hemostatic agent application. Group 3 underwent hemostatic agent application to the renal parenchyma. In Group 4 partial nephrectomy was performed and the hemostatic agent was used without hilar control. Warm ischemia and partial nephrectomy times, and the number of hemostatic agent applications were recorded. Histopathological evaluations were completed. The Fisher, Kruskal Wallis and Mann-Whitney U tests were used for statistical analysis. RESULTS: Mean kidney size was 2 x 2.5 x 0.05 cm. Mean partial nephrectomy time was 3.7, 2.7, 1.8 and 3.2 minutes in groups 1 to 4, respectively, which was significantly different between groups 1 and 3 (p = 0.007). Warm ischemia time in group 3 was less than in group 1 (p = 0.011). The number of hemostatic agent applications was higher in groups 3 and 4 compared to those in group 2 (p = 0.003). Glomerular necrosis was detected at a higher rate in group 1 compared to that in groups 3 and 4 (p = 0.015). Calcification formed significantly more in group 1 than in groups 2 to 4 (p <0.05). Erythrocyte aggregation was greater in groups 2 to 4 than in group 1 (p = 0.015). Giant cell reaction, fibrosis, inflammation and microvascular proliferation were not statistically different among the groups (p >0.05). CONCLUSIONS: Ankaferd BloodStopper decreases partial nephrectomy and warm ischemia times, and provides hemostasis. Erythrocyte aggregation confirmed the hemostatic action of the agent, while the absence of glomerular necrosis and calcification may have positive relevance. PMID- 19303090 TI - Increased urinary neutrophil gelatinase associated lipocalin levels in a rat model of upper urinary tract infection. AB - PURPOSE: Recurrent upper urinary tract infection is a common complication of vesicoureteral reflux that often leads to irreversible renal scarring. In our previous study of a rat model of renal bacterial infection we performed global gene expression profiling of the kidney during the onset of renal scarring. We have further investigated the product of an up-regulated gene product, NGAL, in this animal model to evaluate its potential usefulness as a biomarker of renal scarring. MATERIALS AND METHODS: Renal NGAL mRNA and protein levels were examined by real-time polymerase chain reaction, Western blot and immunohistochemistry. Urinary NGAL levels were monitored by direct enzyme-linked immunosorbent assay. RESULTS: Rat renal NGAL mRNA and protein levels were found to be increased soon after bacterial injection. They then decreased rapidly but subsequently persisted at high levels until the 6-week time point after injection. On histological analysis we found that NGAL protein was overproduced in macrophages and renal tubular cells 2 weeks after injection. However, renal tubular cells continued to produce NGAL 6 weeks after injection, whereas this expression was lost in infiltrating cells. Rat urinary NGAL levels were also markedly increased at the early stages of infection and they persisted at high levels throughout the latter stages of the experiment. CONCLUSIONS: Urinary NGAL may be a potential noninvasive diagnostic biomarker of renal scarring. PMID- 19303091 TI - A ruptured calix: unusual presentation of ureteropelvic junction obstruction in an adolescent male. PMID- 19303092 TI - Hypospadias rates in new york state are not increasing. AB - PURPOSE: The testicular dysgenesis syndrome describes urogenital abnormalities associated with exposure to environmental endocrine disruptors such as phthalates, specifically decreased semen quality, and increased rates of testis cancer and hypospadias. Recently there has been concern that these abnormalities described in animal studies may also be present in humans. To determine if hypospadias rates are increasing, we retrospectively reviewed the total prevalence of hypospadias in New York State from 1992 to 2005, categorized by maternal age younger than 35 years and 35 years or older. MATERIALS AND METHODS: Hypospadias rates were obtained from the New York State Congenital Malformations Registry from 1992 to 2005. An analysis was also performed on the rates of children with hypospadias who had mothers younger than 35 years and mothers 35 years or older. This investigation was approved by the Columbia University internal review board. RESULTS: There was no statistical change in hypospadias rates in New York State from 1992 to 2005 (r = 0.127, p = 0.6). Overall the mean +/- SE prevalence rate was 34.9 +/- 0.36 per 10,000 live births. However, mean +/ SE hypospadias rates in children of mothers 35 years old or older (38.7 +/- 0.7) were significantly greater than those in children of mothers younger than 35 years (34.1 +/- 0.386, t test p <0.01). CONCLUSIONS: Hypospadias rates have not changed in New York State from 1992 to 2005. Additionally advanced maternal age continues to be a risk factor for hypospadias. Combined with previous studies that demonstrate sperm counts are not declining, these data suggest that the testicular dysgenesis syndrome described in animal models may not be evident in humans. PMID- 19303093 TI - Management of erectile function by penile purinergic p2 receptors in the diabetic rat. AB - PURPOSE: We determined the role of purine and pyrimidine nucleotides in erectile function in diabetic rats. MATERIALS AND METHODS: A total of 60 adult male rats were divided into 2 groups, including 30 controls and 30 treated with streptozotocin (60 mg/kg) for 8 weeks to induce hyperglycemia. Changes in intracavernous pressure after intracrural injections of adenosine 5'triphosphate and adenosine 5'triphosphate analogues in control and diabetic rats, and the relaxant response to electrical field stimulation of precontracted corpus cavernosum smooth muscle in organ baths were investigated. The localization of P2X1, P2Y1 and P2Y2 receptors was assessed in penile tissue via an immunohistochemical approach. RESULTS: Corpus cavernosum smooth muscle relaxation in vivo and by electrical field stimulation in vitro was significantly decreased in diabetic rats. Adenosine 5'triphosphate (P2X, P2Y), 2-methylthioadenosine 5'triphosphate (P2Y1) and uridine 5'-triphosphate (P2Y2) agonists but not alpha,beta-methylene adenosine 5'triphosphate (a P2X1 agonist) significantly improved the erectile response to electrical field stimulation in diabetic rat corpus cavernosum smooth muscle. Although intracavernous pressure/mean arterial pressure values in the rats were not restored in the presence of the P2X1 antagonist PPADS, the relaxation response to electrical field stimulation in isolated corpus cavernosum smooth muscle from diabetic rats was improved. Abundant immunoreactivity for PX1 and P2Y2 receptors was observed in penile tissues from diabetic rats compared to that from control rats. CONCLUSIONS: These results demonstrate 1) heterogeneous effects of purinergic agonists on corporeal function in diabetic rats, and 2) the activation of P2Y1 and P2Y2 receptor relaxation of corpus cavernosum smooth muscle to induce erection in rats and perhaps improve erectile function in men with diabetes. PMID- 19303094 TI - Role of K+ channels in regulating spontaneous activity in detrusor smooth muscle in situ in the mouse bladder. AB - PURPOSE: We investigated the functional role of K(+) channels for regulating spontaneous activity in mouse bladder detrusor smooth muscle. MATERIALS AND METHODS: The effects of different K(+) channels blockers on spontaneous changes in membrane potential and intracellular Ca(2+) dynamics were examined using intracellular recording techniques and Ca(2+) imaging with fluo-4 fluorescence, respectively. RESULTS: Detrusor smooth muscle generated spontaneous action potentials and Ca(2+) transients. Iberiotoxin (0.1 microM), charybdotoxin (0.1 microM) or tetraethylammonium (1 mM) increased the amplitude of action potentials and prolonged their repolarizing phase without inhibiting their after hyperpolarization. Tetraethylammonium (10 mM) but not stromatoxin (0.1 microM) suppressed after-hyperpolarization and further increased the amplitude and half duration of action potentials. Apamin (0.1 microM) increased the frequency of action potentials but had no effect on their configuration. Spontaneous Ca(2+) transients were generated in individual detrusor smooth muscle cells and occasionally propagated to neighboring cells to form intercellular Ca(2+) waves. Transmural nerve stimulations invariably initiated synchronous Ca(2+) transients within and across muscle bundles. Charybdotoxin (0.1 microM) increased the amplitude of spontaneous Ca(2+) transients, while the subsequent application of tetraethylammonium (10 mM) increased their half duration. In addition, tetraethylammonium increased the synchronicity of Ca(2+) transients in muscle bundles. CONCLUSIONS: These results suggest that large and intermediate conductance Ca(2+) activated K(+) channels contribute to action potential repolarization and restrict the excitability of detrusor smooth muscle in the mouse bladder. In addition, the activation of voltage dependent K(+) channels is involved in repolarization and after-hyperpolarization, and it has a fundamental role in stabilizing detrusor smooth muscle excitability. PMID- 19303095 TI - Differential expression of KAI1 metastasis suppressor protein in renal cell tumor histological subtypes. AB - PURPOSE: The similar appearance of renal tumor histological subtypes can complicate differential diagnoses. This problem is most notable for the chromophobe subtype of renal cell carcinoma, which can be histologically indistinguishable from oncocytoma with investigational molecular markers failing to provide reliable differentiation. KAI1 is a metastasis suppressor gene whose expression correlates inversely with the metastatic potential of most solid tumor cancer types. We tested the hypothesis that KAI1 is differentially expressed among renal tumor histological subtypes. MATERIALS AND METHODS: Immunohistochemical staining for KAI1 protein was performed in 152 nephrectomy specimens, including 48 clear cell, 35 papillary and 31 chromophobe renal cell carcinoma samples, 28 oncocytomas and 10 tumor-free kidneys. Staining was scored as none/minimal, low, moderate or high. KAI1 mRNA levels were compared by quantitative reverse transcriptase-polymerase chain reaction in an additional 22 chromophobe renal cell carcinoma and oncocytoma samples. RESULTS: In all 10 tumor free kidneys KAI1 protein was detected exclusively in distal tubule cell membranes. Of the tumor specimens KAI1 protein was absent in all papillary renal cell carcinoma specimens. It was present in only 1 of 48 clear cell renal cell carcinomas (2%) and 2 of 28 oncocytomas (7%) but only at low levels. In contrast, 27 of 31 chromophobe renal cell carcinoma specimens (87%) expressed KAI1 protein, most at moderate or high levels. The diagnostic accuracy of KAI1 immunostaining for discerning chromophobe renal cell carcinoma from oncocytoma was 90% with similar results observed at the RNA level. CONCLUSIONS: KAI1 is an accurate biomarker for chromophobe renal cell carcinoma that may aid in the diagnostic differentiation of chromophobe renal cell carcinoma from oncocytoma. It remains to be determined whether KAI1 expression contributes to the low metastatic potential of chromophobe renal cell carcinoma. PMID- 19303096 TI - Tamm-horsfall protein protects against urinary tract infection by proteus mirabilis. AB - PURPOSE: Proteus mirabilis is a common cause of urinary tract infection. We determined the role of Tamm-Horsfall protein as a host defense factor against the cystitis and pyelonephritis caused by P. mirabilis. MATERIALS AND METHODS: We generated Tamm-Horsfall protein gene knockout mice using homologous recombination. We introduced P. mirabilis transurethrally into the bladder of Tamm-Horsfall protein deficient (THP(-/-)) and genetically similar WT (THP(+/+)) mice. We cultured urine to quantitate the degree of bacteriuria. We examined bladders and kidneys grossly and histomorphometrically to determine the intensity of inflammation. RESULTS: THP(-/-) mice had more severe bacteriuria and cystitis than THP(+/+) mice. THP(-/-) mice had more pyelonephritic abscesses than THP(+/+) mice. The severity of histological pyelonephritis on semiquantitative histomorphometric analysis appeared to be greater in THP(-/-) mice. The difference between the 2 groups approached but did not attain statistical significance (p = 0.053). CONCLUSION: Tamm-Horsfall protein acts as a host defense factor against P. mirabilis induced urinary tract infection. PMID- 19303097 TI - Activation of RAS family genes in urothelial carcinoma. AB - PURPOSE: Bladder cancer is the fifth most common malignancy in men in Western society. We determined RAS codon 12 and 13 point mutations and evaluated mRNA expression levels in transitional cell carcinoma cases. MATERIALS AND METHODS: Samples from 30 human bladder cancers and 30 normal tissues were analyzed by polymerase chain reaction/restriction fragment length polymorphism and direct sequencing to determine the occurrence of mutations in codons 12 and 13 of RAS family genes. Moreover, we used real-time reverse transcriptase-polymerase chain reaction to evaluate the expression profile of RAS genes in bladder cancer specimens compared to that in adjacent normal tissues. RESULTS: Overall H-RAS mutations in codon 12 were observed in 9 tumor samples (30%). Two of the 9 patients (22%) had invasive bladder cancer and 7 (77%) had noninvasive bladder cancer. One H-RAS mutation (11%) was homozygous and the remaining 89% were heterozygous. All samples were WT for K and N-RAS oncogenes. Moreover, 23 of 30 samples (77%) showed over expression in at least 1 RAS family gene compared to adjacent normal tissue. K and N-RAS had the highest levels of over expression in bladder cancer specimens (50%), whereas 27% of transitional cell carcinomas demonstrated H-RAS over expression relative to paired normal tissues. CONCLUSIONS: Our results underline the importance of H-RAS activation in human bladder cancer by codon 12 mutations. Moreover, they provide evidence that increased expression of all 3 RAS genes is a common event in bladder cancer that is associated with disease development. PMID- 19303098 TI - Re: Renal mass biopsy--a renaissance? B. R. Lane, M. K. Samplaski, B. R. Herts, M. Zhou, A. C. Novick and S. C. Campbell J Urol 2008; 179: 20-27. PMID- 19303099 TI - Longitudinal development of renal damage and renal function in infants with high grade vesicoureteral reflux. AB - PURPOSE: We sought to study renal abnormality and renal function through time in infants with high grade vesicoureteral reflux. MATERIALS AND METHODS: This prospective observational study included 115 infants (80 boys and 35 girls) younger than 1 year with grade III to V vesicoureteral reflux. The diagnosis was made after prenatal ultrasound in 26% of the patients and after urinary tract infection in 71%. Patients were followed by renal scintigraphy, 51chromium edetic acid clearance and video cystometry. Median followup was 62 months. RESULTS: Renal abnormality, which was found in 90% of the children at followup, was generalized in 71% and focal in 29%. The abnormality was bilateral in 28% of the affected patients. Total glomerular filtration rate was less than 80% of expected in 30% of the patients. Single kidney function was less than 40% of expected total glomerular filtration rate in 71% of the patients. Renal status (parenchymal abnormality and function) remained unchanged through time in 84 of 108 available cases (78%), improved in 5 (5%) and deteriorated in 19 (18%). Predictive factors for deterioration were recurrent febrile urinary tract infection, bilateral abnormality and reduced total glomerular filtration rate. Deteriorated renal status was more common in cases diagnosed prenatally than in those detected after urinary tract infection. CONCLUSIONS: Among these infants with high grade vesicoureteral reflux renal abnormality was frequent and was associated with subnormal filtration of one of the kidneys. Decreased total glomerular filtration rate was seen in about a third of the patients. Overall deterioration of renal status was seen in only a fifth of the patients. Infection control seems to be an important factor to minimize the risk. PMID- 19303101 TI - Primary ablation of posterior urethral valves in low birth weight neonates by a visually guided fogarty embolectomy catheter. AB - PURPOSE: In low birth weight neonates primary ablation of posterior urethral valves represents a particular difficulty. The tiny caliber of the urethra presents a challenge for even the smallest commercially available pediatric resectoscope. Transurethral ablation by wired electrodes is difficult due to the narrow field of neonatal scopes with slow irrigation and restricted maneuverability. A Fogarty embolectomy catheter under fluoroscopic control does not allow direct visual assessment, and can be a lengthy procedure with numerous manipulations at the radiology suite. We describe a hybrid technique for primary valve ablation in neonates using a Fogarty catheter working retrograde under direct visual guidance of a neonatal cystoscope. MATERIALS AND METHODS: A total of 17 low birth weight newborns (median 2,100 gm, range 1,760 to 2,690) underwent primary valve ablation using a 2Fr Fogarty catheter working through a 7.5Fr neonatal cystoscope with an offset lens. Both components were withdrawn as a single unit to avulse the valve leaflets under vision. Vesicoureteral reflux was present in 13 patients (76%) involving 24 renal units. A voiding cystourethrogram was performed 8 to 10 weeks postoperatively to delineate adequate decompression of the posterior urethra and mark the end point of the study in evaluating the efficiency of the technique. RESULTS: There were no immediate perioperative complications. Three patients died of causes unrelated to the procedure, leaving 14 boys (21 refluxing units) for evaluation. Voiding cystourethrogram revealed effective decompression of the posterior urethra with adequate drainage in all but 1 remaining patient (93%) due to a persistent left leaflet. Of 21 refluxing units reflux resolved in 6 (29%), was downgraded in 10 (48%) and remained unchanged in 5 (24%). CONCLUSIONS: In low birth weight neonates primary valve ablation by a visually guided Fogarty catheter ensures effective disruption of the valvular obstructive mechanism. The technique maintains the transurethral visual standard and avoids the difficult manipulation of wired electrodes in a restricted field. It is devoid of radiation exposure and its simplicity enables safe performance at the neonatal intensive care unit, where the support needs of this fragile subset of patients are optimally provided for. PMID- 19303104 TI - Sesquiterpene lactone parthenolide ameliorates bladder inflammation and bladder overactivity in cyclophosphamide induced rat cystitis model by inhibiting nuclear factor-kappaB phosphorylation. AB - PURPOSE: Cyclophosphamide (Sigma) is associated with urological complications, including irritative voiding symptoms and hemorrhagic cystitis. Evidence suggests that tumor necrosis factor-alpha (R&D Systems), interleukin-1beta and cyclooxygenase-2 are directly involved in the pathogenesis of cyclophosphamide induced cystitis and these molecules depend on transcription factor NF-kappaB for maximal secretion. Additionally, sesquiterpene lactone parthenolide has been shown to be a potent nuclear factor-kappaB inhibitor. We hypothesized that enhanced nuclear factor-kappaB activity contributes to cyclophosphamide induced cystitis and, therefore, it may be an attractive target for preventing cyclophosphamide cystitis. We determined whether parthenolide could be used as a preventive agent for hemorrhagic cystitis and bladder overactivity. Moreover, we determined the molecular mechanisms of parthenolide on the inhibitory action of nuclear factor-kappaB in inflammatory human benign urothelial cells. MATERIALS AND METHODS: Rats were pretreated with parthenolide or vehicle solution and administered cyclophosphamide. Histological analysis and cystometry were performed 24 hours after cyclophosphamide administration. Human urothelial cells were pretreated with parthenolide and stimulated with tumor necrosis factor alpha. Western blotting and immunofluorescence were performed to determine activation of the cyclooxygenase-2 and nuclear factor-kappaB pathway. RESULTS: Parthenolide pretreatment inhibited bladder inflammation as well as bladder overactivity and it was also associated with nuclear factor-kappaB activation in the bladder. Parthenolide dose dependently suppressed tumor necrosis factor-alpha induced cyclooxygenase-2 expression and prevented nuclear factor-kappaB phosphorylation as well as nuclear factor-kappaB nuclear translocation and IkappaBalpha phosphorylation/degradation. CONCLUSIONS: Nuclear factor-kappaB may have a crucial role in the pathogenesis of cyclophosphamide induced cystitis models. Parthenolide ameliorates bladder inflammation and bladder overactivity, and it might be a promising agent for preventing cyclophosphamide induced complications. PMID- 19303105 TI - Leydig cell tumor of the testis. PMID- 19303106 TI - Genetic polymorphisms of matrix metalloproteinases: susceptibility and prognostic implications for prostate cancer. AB - PURPOSE: Prostate cancer is the most common tumor in males in Brazil. Single nucleotide polymorphisms have been demonstrated to exist in the promoter regions of matrix metalloproteinase genes and they are associated with the development and progression of some cancers. We investigated the correlation between MMP1, 2, 7 and 9 polymorphisms with susceptibility to prostate cancer, and classic prognostic parameters of prostate cancer. MATERIALS AND METHODS: Genomic DNA was extracted using conventional protocols. The DNA sequence containing the polymorphic site was amplified by real-time polymerase chain reaction using TaqMan(R) fluorescent probes. RESULTS: For the MMP1 gene the polymorphic allele was more common in the control group than in the prostate cancer group (p <0.001). For the MMP9 gene the incidence of the polymorphic homozygote genotype was higher in the prostate cancer group (p <0.001). For higher stage tumors (pT3) a polymorphic allele in the MMP2 gene was more common (p = 0.026). When considering Gleason score, the polymorphic homozygote genotype of MMP9 was more common in Gleason 6 or less tumors (p = 0.003), while a polymorphic allele in the MMP2 gene was more common in Gleason 7 or greater tumors (p = 0.042). CONCLUSIONS: MMP1 and MMP2 may protect against prostate cancer development and MMP9 may be related to higher risk. In contrast, MMP9 polymorphism was associated with a lower Gleason score and MMP2 polymorphism was associated with nonorgan confined disease. PMID- 19303107 TI - Unraveling the genetic landscape of bladder development in mice. AB - PURPOSE: To better understand the pathobiology of human congenital bladder abnormalities and disorders associated with dedifferentiation, such as bladder cancer, we must first unravel the biology of normal bladder development. Therefore, we performed microarray analysis focusing on determining the gene expression profile at the initiation of bladder development. MATERIALS AND METHODS: RNA was extracted from embryonic day 13 and 18 mouse bladders (anatomically equivalent to 7 and 13 weeks of human gestation) and gene expression was evaluated using microarrays. Alterations in select genes of biological interest were confirmed using real-time quantitative polymerase chain reaction and localization was determined by immunohistochemistry. RESULTS: The genetic profile in the initiating mouse bladder at embryonic day 13 was dominated by transcription factors, retinoic acid signaling genes, Eph/ephrin bidirectional signaling molecules and genes associated with regulating cell cycle and differentiation. Later in development at embryonic day 18 genes associated with smooth muscle, innervation and epithelial differentiation were up-regulated. In addition, we examined the functional role of midkine, which was highly expressed at embryonic day 13, using organ culture and to our knowledge we provide the first evidence that this growth factor up-regulates molecules associated with bladder smooth muscle differentiation. CONCLUSIONS: These data provide novel insights into molecules that orchestrate bladder development and highlight genes that may be involved in diseases associated with abnormal differentiation. PMID- 19303111 TI - The amphipods Caprella penantis and Hyale schmidtii as biomonitors of trace metal contamination in intertidal ecosystems of Algeciras Bay, Southern Spain. PMID- 19303112 TI - Geographical variability of the incidence of Type 1 diabetes in subjects younger than 30 years in Catalonia, Spain. AB - BACKGROUND AND OBJECTIVE: We decided to assess the geographical variability of the incidence of Type 1 diabetes in Catalonia (Spain) in subjects younger than 30 years at onset during the period 1989-1998. The effect of sex, age at onset, periods of years, and population density was also analyzed. MATERIAL AND METHODS: Data were obtained from the prospective Catalan Registry of Diabetes Mellitus. Generalized linear mixed models were used to determine the effects of the risk factors and to find out the geographical distribution. The best model was selected by the AKAIKE information criterion. RESULTS: The crude incidence of type 1 diabetes in subjects younger than 30 years was 11.8/100,000/year (95% CI 11.4-12.3). The incidence was similar between males and females in the 0-14 age group. However, there was a male preponderance in young adults. The incidence did not vary annually and was not associated with population density. The incidence did not present a spatial pattern around Catalonia. There was an unstructured geographical variability. CONCLUSIONS: Some regions of Catalonia displayed values of type I diabetes higher or lower than the expected incidence. Counties with extreme values of incidence were specific for each demographic group and in no case did these counties make up clusters, suggesting that there are explanatory factors with patterns of geographic distribution. The incidence of diabetes in young male adults in some counties was similar to that of European countries with a high incidence. PMID- 19303113 TI - [Imaging anatomy of cranial nerves]. AB - Knowledge of the anatomy of the cranial nerves is mandatory for optimal radiological exploration and interpretation of the images in normal and pathological conditions. CT is the method of choice for the study of the skull base and its foramina. MRI explores the cranial nerves and their vascular relationships precisely. Because of their small size, it is essential to obtain images with high spatial resolution. The MRI sequences optimize contrast between nerves and surrounding structures (cerebrospinal fluid, fat, bone structures and vessels). This chapter discusses the radiological anatomy of the cranial nerves. PMID- 19303114 TI - [Treatment of trigeminal neuralgia with thermorhizotomy]. AB - Percutaneous radiofrequency (RF) thermorhizotomy of the trigeminal nerve is an effective treatment for trigeminal neuralgia. Long-term efficacy is proportional to the degree of postoperative hypoesthesia. The advantage is the topographic selectivity of the thermolesion, provided the electrode tip is placed accurately. This requires precise x-ray guidance and neurophysiological testing. In addition to the indication in idiopathic trigeminal neuralgia, especially for elderly patients with precarious conditions, thermorhizotomy is particularly useful for treating patients with trigeminal neuralgia due to multiple sclerosis. It can also be applied to symptomatic neuralgias, but only when the main components are of the paroxysmal and/or the allodynic types ; the thermorhizotomy method could aggravate permanent components, especially when burning pain predominates; it could also increase preexisting trophic disturbances, particularly keratitis. PMID- 19303115 TI - [Vertigo treatment according to their mechanisms]. AB - Vertigo is an illusion of rotatory or linear movement that demonstrates a functional or lesional disturbance of the vestibular system, from periphery to central connections. According to the ANAES report (1997), benign paroxysmal positional vertical vertigo, vestibular neuronitis and Meniere's disease account for 40-50% of all mixed vertigo etiologies. Central etiologies may account for 20 40% of causes and 10-40% remain more difficult to classify, and are usually classified under the term of "peripheral vestibulopathy." These include vertigo due to neurovascular compression syndrome of the VIIIth nerve. Clinical manifestations, differential diagnosis, and treatment of the main etiologies of vertigo will be developed in this chapter. A specific section will discuss the subject of neurovascular compression syndrome of the VIIIth nerve. Even though some publications should be challenged, it appears that neurovascular compression syndrome of the VIIIth nerve might explain some cases of vertigo or chronic instability, with or without cochlear signs. The diagnosis is difficult and must be established on multiple clinical, electrophysiological and radiological arguments. A therapeutic test with antiepileptic drugs is helpful. The treatment includes these drugs as a first option but may require a neurosurgical approach if medical treatment fails. PMID- 19303116 TI - [Treatment of trigeminal neuralgia with glycerol injection at the gasserian ganglion]. AB - Gasserian ganglion neurolysis with glycerol injected percutaneously through the foramen ovale continues to be widely used. Its long-term efficacy on pain is proportional to the degree of postoperative hypoesthesia. The advantage is low cost. The disadvantage is essentially the difficulty in controlling diffusion outside the Meckel cavity. Subsequently its effects are somewhat random and potential complications are difficult to prevent reliably. PMID- 19303117 TI - [Functional anatomy of the trigeminal nerve]. AB - The cranial nerve (CN) V is a mixed nerve that consists primarily of sensory neurons. It exits the brain on the lateral surface of the pons, entering the trigeminal ganglion within a few millimeters. Three major branches emerge from the trigeminal ganglion. The first division (V1, the ophthalmic nerve) exits the cranium through the superior orbital fissure, entering the orbit to innervate the globe and skin in the area above the eye and forehead. The second division (V2, the maxillary nerve) exits through a round hole, the foramen rotundum, into a space posterior to the orbit, the pterygopalatine fossa. It then re-enters a canal running inferior to the orbit, the infraorbital canal, and exits through a small hole, the infraorbital foramen, to innervate the skin below the eye and above the mouth. The third division (V3, the mandibular nerve) exits the cranium through an oval hole, the foramen ovale. The third division also has an additional motor component, which may run in a separate fascial compartment. Most fibers travel directly to their target tissues. Sensory axons innervate skin on the lateral side of the head, the tongue, and the mucosal wall of the oral cavity. Motor fibers innervate the muscles that are attached to the mandible. Some sensory axons enter in the mandible to innervate the teeth and emerge from the mental foramen to innervate the skin of the lower jaw. PMID- 19303118 TI - [Surgery for vertigo]. AB - Spontaneous recovery or central compensation makes surgical procedures rare in patients with vertigo. Surgery for vertigo proposed after pharmacological or physical therapy fails to eliminate Meniere's disease and some very rare cases of paroxystic positional vertigo. The main target in treating Meniere's disease is to promote vestibular compensation, which is possible only with a nonprogressive and stable deficit leading to readjustment of vestibular reflexes. Surgical procedures can be classified as nondestructive (endolymphatic sac decompression, vestibular nerve decompression, patching of perilymphatic fistulas), selectively destructive (middle fossa or retrosigmoid vestibular neurotomy, lateral semi circular plugging) and destructive (labyrinthectomy). Surgical indications essentially concern incapacitating vertigo and depend mainly on hearing status. In Meniere's disease, vestibular neurotomy can be regarded as the gold standard considering its good results on vertiginous episodes; however, scoring with functional and quality-of-life scales bring out residual deficiency in some cases. PMID- 19303119 TI - English health profiles--did they do what was expected? An evaluation of Health Profiles 2006. AB - OBJECTIVES: To assess the impact of Health Profiles 2006 by English local authorities, and to determine what changes need to be made to the profiles to have an impact on their target audience. STUDY DESIGN: A telephone- and web-based survey of a sample of the health profiles' target audience was conducted, along with an analysis of web statistics. METHODS: In total, 285 telephone interviews were undertaken. Fifty-three percent of the respondents were National Health Service employees. An evaluation form was also available through the Health Profiles website, which elicited 117 responses (19 scrutiny officers and 83 members of the public). RESULTS: There was a positive response to the content and format of Health Profiles 2006. The majority of respondents felt that the profiles provided a unique summary of local-authority-based health and health inequality information that was both accessible and understandable. CONCLUSIONS: A number of recommendations are made to improve health profile production as a tool for information for health improvement. These include simplifying graphics, greater local input into commentary, and a more interactive website so that the data can be accessed and explored. However, the recommendations for change must be viewed alongside the number of comments specifically relating to the English health profiles that nothing should be changed other than updating the data and filling in the gaps. PMID- 19303120 TI - Use of starting condition score to estimate changes in body weight and composition during weight loss in obese dogs. AB - Prior to starting a weight loss programme, target weight (TW) is often estimated, using starting body condition score (BCS). The current study assessed how well such estimates perform in clinical practice. Information on body weight, BCS and body composition was assessed before and after weight loss in 28 obese, client owned dogs. Median decrease in starting weight per BCS unit was 10% (5-15%), with no significant difference between dogs losing moderate (1-2 BCS points) or marked (3-4 BCS points) amounts of weight (P=0.627). Mean decrease in body fat per BCS unit change was 5% (3-9%). A model based on a change of 10% of starting weight per unit of BCS above ideal (5/9) most closely estimated actual TW, but marked variability was seen. Therefore, although such calculations may provide a guide to final TW in obese dogs, they can either over- or under-estimate the appropriate end point of weight loss. PMID- 19303121 TI - Quantification of water buffalo (Bubalus bubalis) cytokine expression in response to inactivated foot-and-mouth disease (FMD) vaccine. AB - This study describes the quantification of cytokine expression of vaccinated water buffaloes with FMD inactivated vaccine. Using real-time PCR quantification assay, expression of Th1 (IL-2, IL-12p40, IFNgamma); Th2 (IL-4, IL-10) and inflammatory (IL-6, TNFalpha) cytokines were quantified weekly for the entire three-week duration of the experiment. It was noted that IFNgamma, IL-10 and TNFalpha had peaked on week three post-vaccination while the remaining cytokines peaked on the second week and decreased by the third week. The counteraction between IFNgamma and IL-4 was noted as well as the possible suppressive action of IL-10 to that of IL-2 and IL-12, which is a common phenomenon between Th1 and Th2 cytokines. Synergy between TNFa and IL-6 was also observed. These findings suggest that within the immune system of water buffalo there is a dynamic cell mediated and humoral interaction in response to immunogen. This assessment of the cytokine expressions is vital for the study of water buffalo disease progression and concurring protective immune responses. PMID- 19303122 TI - Behavioural effects of ovariohysterectomy and oral administration of meloxicam in laboratory housed rabbits. AB - This study aimed to develop a behaviour-based pain assessment system for rabbits following ovariohysterectomy. Behaviour was analysed to assess the severity and duration of pain induced and determine the effects of administration of meloxicam. The results suggest that pain associated with ovariohysterectomy induced changes in the frequency and duration of a number of behaviours. The most indicative was inactive pain behaviour, which was observed very infrequently prior to surgery compared to very frequently immediately following surgery. This strongly suggests that this increase is a direct response to the surgical pain and/or stress. The frequency of inactive pain behaviour also decreased over the four days post-surgery suggesting that pain is decreasing during this time. High dose meloxicam (initial 1mg/kg followed 0.5mg/kg/day) induced some degree of analgesia. However, higher doses of meloxicam or in combination with an opioid may be required to provide consistent analgesia in rabbits following soft-tissue surgery. PMID- 19303123 TI - Intestinal mucosal immune response against virulent duck enteritis virus infection in ducklings. AB - Duck virus enteritis (DVE) is an acute and contagious herpes virus infection of duck, geese and swans with high morbidity and mortality. The development of specific mucosal immune system against duck enteritis virus (DEV) infection for ducks has been hindered by a lack of knowledge concerning the purification of immunoglobulin A (IgA) of duck. In the present work, the method for purification of duck immunoglobulin A was developed, and the induction of intestinal mucosal immune responses against DEV was studied by orally infected ducklings with virulent DEV. The results showed that a continuous increased DEV DNA levels were observed in blood and various organs examined of orally infected ducklings throughout the infection, which was accompanied by the development of infection in ducklings from mild progressed to severe pathological lesions. Furthermore, a marked increased level of DEV-specific IgA and IgG antibodies in bile, serum and the intestinal tract, as well as the density of IgA(+) cells in intestine were detected between 1 and 12days p.i., followed by a drastic reduction of the antibody levels and the density of IgA(+) cells at 15days p.i. The results indicate that the DVE infection can stimulate both IgA-dominated antibody immune responses in the intestinal tract, and IgG-dominated antibody systemic immunity in the serum of ducklings orally inoculated with virulent DEV. The severe lesions of the villus epithelial cells and the lymphoid organs can suppress the intestinal mucosal immune responses. PMID- 19303124 TI - Molecular markers for Phlebotomus papatasi (Diptera: Psychodidae) and their usefulness for population genetic analysis. AB - Three molecular typing tools: multilocus microsatellite typing, cytochrome b sequence analysis and internal transcribed spacer 2 (ITS2) sequence analysis, were evaluated for their usefulness in inferring the population structure of Phlebotomus papatasi sand flies. ITS2 sequence analysis did not prove suitable for inferring phylogenetic and population genetic relationships across P. papatasi sand flies. Microsatellite markers showed high resolution in differentiating globally distributed P. papatasi populations, whereas cytochrome b sequence analysis provided insight into the relationships between closely related populations from the Mediterranean. Population structure, differentiation and demographic history among P. papatasi are important for understanding patterns of dispersal in this species and for planning appropriate control measures. PMID- 19303125 TI - Insecticide resistance and its association with target-site mutations in natural populations of Anopheles gambiae from eastern Uganda. AB - Insecticide resistance in Anopheles gambiae threatens the success of malaria vector control programmes in sub-Saharan Africa. In order to manage insecticide resistance successfully, it is essential to assess continuously the target mosquito population. Here, we collected baseline information on the distribution and prevalence of insecticide resistance and its association with target-site mutations in eastern Uganda. Anopheles gambiae s.l. adults were raised from wild caught larvae sampled from two ecologically distinct breeding sites and exposed to WHO discriminating concentrations of DDT, permethrin, deltamethrin, bendiocarb and malathion. Survival rates to DDT were as high as 85.4%, alongside significant resistance levels to permethrin (38.5%), reduced susceptibility to deltamethrin, but full susceptibility to bendiocarb and malathion. Using molecular diagnostics, susceptible and resistant specimens were further tested for the presence of knockdown resistance (kdr) and acetylcholinesterase 1 resistance (ace-1(R)) alleles. While ace-1(R) and kdrL1014F ('kdr west') alleles were absent, the kdr L1014S ('kdr east') allele was present in both populations. In A. gambiae s.s., L1014S was closely associated with DDT and, to a lesser degree, with permethrin resistance. Intriguingly, the association between DDT resistance and the presence of L1014S is consistent with a co-dominant effect, with heterozygous individuals showing an intermediate phenotype. PMID- 19303126 TI - Microbiological surveillance of private water supplies in England: the impact of environmental and climate factors on water quality. AB - A passive surveillance system captured information on 34,904 microbiological samples from 11,233 private drinking water supplies within England as well as the associated constructional, climatic and environmental variables. Escherichia coli was detected in 6588 (18.87%) of samples and at least one positive sample was detected from 3638 (32.39%) of sites. However, this estimate of supplies failing to meet the European drinking water E. coli standard was probably an underestimate as the more samples taken per supply, the more likely the supply was to fail. A multivariable model of private water supplies data showed a strong seasonal impact, with samples between January and May being significantly less contaminated with E. coli than samples between June and December. Samples from springs (OR 2.5, CI 2.0-3.1) or surface waters (OR 2.4, CI 0.8-7.0) were more likely to fail than groundwater sources, as were supplies with no effective treatment (OR1.8, CI 1.5-2.3). Commercial supplies were less likely to fail than domestic supplies (OR 0.63, CI 0.48-0.83) and the probability of failure was linearly associated with the density of sheep in the area and rainfall on the previous day. A Monte Carlo modelling approach was used to estimate that, had sufficient samples been taken, 54% (95% confidence intervals 49-59%) of all private water supplies in England were likely to be unsatisfactory. These findings will be able to inform risk assessments of private water supplies prior to microbiological results being available. PMID- 19303127 TI - Rejection of pharmaceutically active compounds and endocrine disrupting compounds by clean and fouled nanofiltration membranes. AB - Rejections of 9 pharmaceuticals and 5 endocrine disruptors by clean and fouled nanofiltration membranes were investigated in this study. Waters containing a cocktail of compounds were filtered by clean and pre-fouled membranes. The rejection of hydrophilic neutral compounds by the clean NF-200 membrane varied from 35 to 70% under steady state conditions while that of NF-90 membrane was in the range of 62-96%. The clean NF-90 membrane rejected nearly all of the hydrophobic neutral compounds (95-98%) predominantly due to size exclusion. Nevertheless, electrostatic repulsion was the main mechanism of rejection of ionic compounds by both membranes (71-94% by NF-200 and 99% by NF-90). Fouling with sodium alginate deteriorated the performance of the NF-200 membrane in rejecting hydrophilic neutral compounds as well as hydrophilic and hydrophobic ionic compounds. In contrast, rejections of hydrophobic neutral compounds by the fouled NF-200 membrane increased by 5-38%. This may be attributed to the incipient interaction of the solutes with the membrane foulant layer resulting in less partitioning and diffusion across the membrane surface. On the other hand, rejections of hydrophobic compounds by NF-90 were not observed to be affected by fouling; however, hydrophilic neutral compounds showed increased rejections by 7 30%. PMID- 19303128 TI - Assessment of annual pollutant loads in combined sewers from continuous turbidity measurements: sensitivity to calibration data. AB - This article presents a methodology for assessing annual wet weather Suspended Solids (SS) and Chemical Oxygen Demand (COD) loads in combined sewers, along with the associated uncertainties from continuous turbidity measurements. The proposed method is applied to data from various urban catchments in the cities of Paris and Nantes. The focus here concerns the impact of the number of rain events sampled for calibration (i.e. through establishing linear SS/turbidity or COD/turbidity relationships) on the uncertainty of annual pollutant load assessments. Two calculation methods are investigated, both of which rely on Monte Carlo simulations: random assignment of event-specific calibration relationships to each individual rain event, and the use of an overall relationship built from the entire available data set. Since results indicate a fairly low inter-event variability for calibration relationship parameters, an accurate assessment of pollutant loads can be derived, even when fewer than 10 events are sampled for calibration purposes. For operational applications, these results suggest that turbidity could provide a more precise evaluation of pollutant loads at lower cost than typical sampling methods. PMID- 19303129 TI - Occurrence of dissolved and particle-bound taste and odor compounds in Swiss lake waters. AB - The occurrence of algal taste and odor (T&O) compounds was investigated in three Swiss lakes which exhibit different nutrient levels from eutrophic to oligotrophic (Lake Greifensee, Lake Zurich and Lake Lucerne). Apart from dissolved T&O compounds, the study also encompassed particle-bound compounds, i.e., compounds that can be released from damaged algal cells during drinking water treatment. A combined instrumental (SPME-GC-MS) and sensory method was applied that allowed to detect and quantify T&O compounds in natural waters in the sub ppt to low ppt-range. In addition to the prominent T&O compounds geosmin and 2-methyl-isoborneol (MIB), four other T&O compounds could be detected in the lake waters, though all at relatively low concentrations (maximum concentrations of geosmin 19 ng L(-1), MIB 3 ng L(-1), beta-ionone 27 ng L(-1), beta-cyclocitral 7 ng L(-1), 2-isobutyl-3-methoxypyrazine 2 ng L(-1), 2-isopropyl-3 methoxypyrazine 16 ng L(-1)). The concentration peaks typically occurred in the epilimnion during summer concurrent with a high phytoplankton biomass. Consistently, the concentration levels for most of the compounds varied substantially between the three lakes and generally decreased in the order eutrophic Lake Greifensee>mesotrophic Lake Zurich>oligotrophic Lake Lucerne. Furthermore, our data revealed that the occurrence of beta-ionone was largely influenced by Planktothrix rubescens. This is the first time that a correlation between beta-ionone and this cyanobacterium has been reported for natural waters. PMID- 19303130 TI - Electrochemical degradation of perchloroethylene in aqueous media: an approach to different strategies. AB - An approaching study to the electrochemical degradation of perchloroethylene (PCE) in water has been carried out using controlled current density degradation electrolyses. The different electrochemical strategies to degrade perchloroethylene in aqueous media (i.e. cathodic, anodic and dual treatments) have been checked using divided and undivided configurations. The influence of the initial concentration, pH and current density on the general behavior of the system has been studied, and special attention was paid to the nature of the byproducts formed and to the analysis of the closed mass balance at the end of the reaction. Results from several analytical techniques have been compared. Undivided configuration provides the best results in these experimental conditions, with degradation percentages higher than 50% and with only 6% of the initial perchloroethylene concentration remaining in the system. PMID- 19303131 TI - Controversies about the occurrence of chloral hydrate in drinking water. AB - Besides trihalomethanes (THMs) and haloacetic acids (HAAs), chloral hydrate (CH) is the next most prevalent disinfection by-product (DBP) in drinking water, formed as a result of the reaction between chlorine and natural organic matter (NOM). Chloral hydrate (trichloroacetaldehyde) should be limited in drinking water because of its adverse health effect. The controversies concerning the appearance of CH in disinfected water found in literature are discussed in the present paper. According to some authors the CH yield during chlorination of water depends only on TOC. However, there are other data available that do not confirm this relationship. Another fact requiring clarification is the dependence of CH formation on pH. In the present study, CH formation is analysed in different types of water disinfected with different doses of chlorine. Formation of CH is correlated with the dose of Cl(2) and the contact time. The formation of chloral hydrate takes place as long as chlorine is available in the water. Total organic carbon (TOC) is not considered the main factor influencing the production of chloral hydrate in water treated with Cl(2) as the production depends also on the nature of NOM. Higher levels of CH are observed at alkaline conditions (pH>7). A significant correlation (R(2)>0.9) between the concentrations of chloral hydrate and chloroform has been observed. The preozonation increases significantly the chloral hydrate formation potential in the water treated. Biofiltration process does not remove all of CH precursors and its efficiency depends strongly on the contact time. Chloral hydrate was analyzed by gas chromatography with electron capture detector with the detection limit 0.1 microg L(-1). PMID- 19303132 TI - Photoinitiated oxidation of geosmin and 2-methylisoborneol by irradiation with 254 nm and 185 nm UV light. AB - The degradation of geosmin and 2-methylisoborneol (2-MIB) by UV irradiation at different wavelengths was investigated under varying boundary conditions. The results showed that conventional UV radiation (254 nm) is ineffective in removing these compounds from water. In contrast to the usual UV radiation UV/VUV radiation (254+185 nm) was more effective in the removal of the taste and odour compounds. The degradation could be described by a simple pseudo first-order rate law with rate constants of about 1.2 x 10(-3) m(2)J(-1) for geosmin and 2-MIB in ultrapure water. In natural water used for drinking water abstraction the rate constants decreased to 2.7 x 10(-4) m(2)J(-1) for geosmin and 2.5 x 10(-4) m(2)J( 1) for 2-MIB due to the presence of NOM. Additionally, the formation of the by product nitrite was studied. In the UV/VUV irradiation process up to 0.6 mg L(-1) nitrite was formed during the complete photoinitiated oxidation of the odour compounds. However, the addition of low ozone doses could prevent the formation of nitrite in the UV/VUV irradiation experiments. PMID- 19303133 TI - Effects of ultrasound on suspended particles in municipal wastewater. AB - The objective of this research is to explore the fundamental characteristics of how particles in wastewater respond to ultrasound, with an aim to improve wastewater disinfection. Particles of a predetermined size fraction and concentration were treated with varying doses of ultrasound at 20.3 kHz. Ultrasonic power transfer to the fluid was measured using calorimetry or acoustical measurements. Image analysis particle counting was used to measure the size distribution of particles before and after ultrasound treatment. The influence of three parameters: particle origin (raw wastewater or from the aeration basin of the activated sludge process), particle concentration, and particle size on the percentage of particle breakage after ultrasound treatment was compared. It was found that raw wastewater and aeration basin particles of the same size fraction (90-106 microm) responded to ultrasound in a similar way. Particle breakage was not affected by changes in particle concentration from 100 to 400 particles per mL. Larger wastewater particles (90-250 microm) were more susceptible to breakage than smaller ones (38-63 microm diameter). The percentage of particle breakage increased linearly with a logarithmic increase in the ultrasound energy density, that is the ultrasound energy delivered per unit volume of the sample (R(2)=0.48-0.91). An expression that predicts the percent of particles broken as a function of ultrasound energy density is provided. PMID- 19303134 TI - Levels of six estrogens in water and sediment from three rivers in Tianjin area, China. AB - The occurrence of estrogens in the aquatic environment attracts increasing attention because of their strong endocrine disrupting potency. In the present work, concentrations of six estrogens including diethylstilbestrol (DES), estrone (E1), beta-estradiol (E2), estriol (E3), 17alpha-ethynylestradiol (EE2) and beta estradiol 17-valerate (EV) in surface water and sediment sampled from three rivers in Tianjin area, northern China, were determined by gas chromatography mass spectrometry (GC-MS). The concentrations of all six estrogens (Sigma6ES) ranged from 0.64 to 174 ng L(-1) in waters and from 0.98 to 51.6 ng g(-1) dry weight (dw) in sediments, and varied for each river. Among these estrogens, E1 was the most abundant and could be detected in all samples. DES and EV could be detected either in river water or in sediment, but in the concentration below 10 ng L(-1) and 10 ng g(-1) for water and sediment, respectively. The relationships between concentration of estrogens and organic carbon content in sediments and the relationships between sediment-water partition coefficient (logK(oc)) and octanol-water partition coefficient (logK(ow)) were examined. The results showed that the contents of the Sigma6ES correlated significantly with the contents of organic carbon (OC%). It indicated that sediments with high organic carbon were more likely to retain estrogens than those with lower organic carbon levels in the natural aquatic environment. Furthermore, the linear correlations between logK(ow) and logK(oc) were obtained for each river, which indicated that sediment water partitioning of estrogens in three rivers could be predicted by their hydrophobic properties. PMID- 19303135 TI - Influence of soil acidification in southern Norway on the 137Cs exposure of moose? AB - The mean (137)Cs activity concentration in 278 liver samples of moose (Alces alces) from 16 municipalities located in different parts of Norway varied within the range 43-752 Bq kg(-1) among the municipalities. In general the geographical variation corresponded to the fallout pattern produced by the Chernobyl accident. In three communities in the southernmost part of the country however the transfer factor, defined as the activity in moose liver divided by the corresponding level in surface soil, was 6.5 times higher on average than elsewhere in Norway. Possible reasons for this highly significant difference are discussed, and it is hypothesized that the apparently much higher plant uptake in the south may be related to extensive soil acidification in this area from transboundary pollution. PMID- 19303136 TI - Polycomb proteins remain bound to chromatin and DNA during DNA replication in vitro. AB - The transcriptional status of a gene can be maintained through multiple rounds of cell division during development. This epigenetic effect is believed to reflect heritable changes in chromatin folding and histone modifications or variants at target genes, but little is known about how these chromatin features are inherited through cell division. A particular challenge for maintaining transcription states is DNA replication, which disrupts or dilutes chromatin associated proteins and histone modifications. PRC1-class Polycomb group protein complexes are essential for development and are thought to heritably silence transcription by altering chromatin folding and histone modifications. It is not known whether these complexes and their effects are maintained during DNA replication or subsequently re-established. We find that when PRC1-class Polycomb complex-bound chromatin or DNA is replicated in vitro, Polycomb complexes remain bound to replicated templates. Retention of Polycomb proteins through DNA replication may contribute to maintenance of transcriptional silencing through cell division. PMID- 19303137 TI - Adjuvant imatinib mesylate after resection of localised, primary gastrointestinal stromal tumour: a randomised, double-blind, placebo-controlled trial. AB - BACKGROUND: Gastrointestinal stromal tumour is the most common sarcoma of the intestinal tract. Imatinib mesylate is a small molecule that inhibits activation of the KIT and platelet-derived growth factor receptor alpha proteins, and is effective in first-line treatment of metastatic gastrointestinal stromal tumour. We postulated that adjuvant treatment with imatinib would improve recurrence-free survival compared with placebo after resection of localised, primary gastrointestinal stromal tumour. METHODS: We undertook a randomised phase III, double-blind, placebo-controlled, multicentre trial. Eligible patients had complete gross resection of a primary gastrointestinal stromal tumour at least 3 cm in size and positive for the KIT protein by immunohistochemistry. Patients were randomly assigned, by a stratified biased coin design, to imatinib 400 mg (n=359) or to placebo (n=354) daily for 1 year after surgical resection. Patients and investigators were blinded to the treatment group. Patients assigned to placebo were eligible to crossover to imatinib treatment in the event of tumour recurrence. The primary endpoint was recurrence-free survival, and analysis was by intention to treat. Accrual was stopped early because the trial results crossed the interim analysis efficacy boundary for recurrence-free survival. This study is registered with ClinicalTrials.gov, number NCT00041197. FINDINGS: All randomised patients were included in the analysis. At median follow-up of 19.7 months (minimum-maximum 0-56.4), 30 (8%) patients in the imatinib group and 70 (20%) in the placebo group had had tumour recurrence or had died. Imatinib significantly improved recurrence-free survival compared with placebo (98% [95% CI 96-100] vs 83% [78-88] at 1 year; hazard ratio [HR] 0.35 [0.22-0.53]; one sided p<0.0001). Adjuvant imatinib was well tolerated, with the most common serious events being dermatitis (11 [3%] vs 0), abdominal pain (12 [3%] vs six [1%]), and diarrhoea (ten [2%] vs five [1%]) in the imatinib group and hyperglycaemia (two [<1%] vs seven [2%]) in the placebo group. INTERPRETATION: Adjuvant imatinib therapy is safe and seems to improve recurrence-free survival compared with placebo after the resection of primary gastrointestinal stromal tumour. FUNDING: US National Institutes of Health and Novartis Pharmaceuticals. PMID- 19303138 TI - Adjuvant imatinib in GIST: a self-fulfilling prophecy, or more? PMID- 19303139 TI - N-acetyl cysteine (NAC)-mediated detoxification and functionalization of poly(methyl methacrylate) bone cement. AB - Currently used poly(methyl methacrylate) (PMMA)-based bone cement lacks osteoconductivity and induces osteolysis and implant loosening due to its cellular and tissue-toxicity. A high percentage of revision surgery following the use of bone cement has become a significant universal problem. This study determined whether incorporation of the amino acid derivative N-acetyl cysteine (NAC) in bone cement reduces its cytotoxicity and adds osteoconductivity to the material. Biocompatibility and bioactivity of PMMA-based bone cement with or without 25mm NAC incorporation was examined using rat bone marrow-derived osteoblastic cells. Osteoconductive potential of NAC-incorporated bone cement was determined by microCT bone morphometry and implant biomechanical test in the rat model. Generation of free radicals within the polymerizing bone cement was examined using electron spin resonance spectroscopy. Severely compromised viability and completely suppressed phenotypes of osteoblasts on untreated bone cement were restored to the normal level by NAC incorporation. Bone volume formed around 25mm NAC-incorporated bone cement was threefold greater than that around control bone cement. The strength of bone-bone cement integration was 2.2 times greater for NAC-incorporated bone cement. For NAC-incorporated bone cement, the spike of free radical generation ended within 12h, whereas for control bone cement, a peak level lasted for 6 days and a level greater than half the level of the peak was sustained for 20 days. NAC also increased the level of antioxidant glutathione in osteoblasts. These results suggest that incorporation of NAC in PMMA bone cement detoxifies the material by immediate and effective in situ scavenging of free radicals and increasing intracellular antioxidant reserves, and consequently adds osteoconductivity to the material. PMID- 19303140 TI - Human placental adenosine receptor expression is elevated in preeclampsia and hypoxia increases expression of the A2A receptor. AB - Placental hypoxia as a result of impaired trophoblast invasion is suggested to be involved in the pathophysiology of preeclampsia. Hypoxia is a potent stimulus for the release of adenosine, and the actions of adenosine are mediated through four adenosine receptors, A(1), A(2A), A(2B) and A(3). We investigated the presence, distribution and expression of adenosine receptor subtypes in the human placenta, the expression of the adenosine receptors in placentas from pregnancies complicated by preeclampsia, small for gestational age (SGA) infants and uncomplicated pregnancies, and the effect of hypoxia on placental adenosine receptor expression. Immunofluorescent microscopy localized A(1), A(2A), A(2B) and A(3) adenosine receptors to the syncytiotrophoblast, endothelial cells and myofibroblasts within the human placenta. Adenosine receptor protein and message expression levels were significantly higher in placentas from preeclamptic pregnancies with or without SGA infants, but not different in pregnancies with SGA infants alone. In vitro exposure of placental villous explants to hypoxia (2% oxygen) increased the expression of A(2A) adenosine receptor 50%. These data indicate that all four known adenosine receptors are expressed in the human placenta and adenosine receptor expression is significantly higher in pregnancies complicated by preeclampsia. These data are consistent with the hypothesis that differences in placental adenosine receptors may contribute to alterations in placental function in preeclampsia. PMID- 19303141 TI - Children's adjustment problems in families characterized by men's severe violence toward women: does other family violence matter? AB - OBJECTIVE: This research examined whether additional forms of family violence (partner-child aggression, mother-child aggression, and women's intimate partner violence [IPV]) contribute to children's adjustment problems in families characterized by men's severe violence toward women. METHODS: Participants were 258 children and their mothers recruited from domestic violence shelters. Mothers and children completed measures of men's IPV, women's IPV, partner-child aggression, and mother-child aggression. Mothers provided reports of children's internalizing and externalizing behavior problems; children provided reports of their appraisals of threat in relation to interparent conflict. RESULTS: After controlling for sociodemographics and men's IPV: (1) each of the additional forms of family violence (partner-child aggression, mother-child aggression, and women's IPV) was associated with children's externalizing problems; (2) partner child aggression was associated with internalizing problems; and (3) partner child aggression was associated with children's threat appraisals. The relation of mother-child aggression to externalizing problems was stronger for boys than for girls; gender differences were not observed for internalizing problems or threat appraisals. CONCLUSIONS: Men's severe IPV seldom occurs in the absence of other forms of family violence, and these other forms appear to contribute to children's adjustment problems. Parent-child aggression, and partner-child aggression in particular, are especially important. Systematic efforts to identify shelter children who are victims of parental violence seem warranted. PRACTICE IMPLICATIONS: Men's severe IPV seldom occurs in the absence of other forms of family violence (partner-child aggression, mother-child aggression, and women's IPV), and these different forms of family violence all contribute to children's adjustment problems. Treatment programs for children who come to domestic violence shelters should address these different forms of family violence, especially parent-child aggression. PMID- 19303142 TI - A yeast assay based on the gilthead sea bream (teleost fish) estrogen receptor beta for monitoring estrogen mimics. AB - A yeast (Saccharomyces cerevisiae)-based assay was developed and tested with steroids and chemicals (mostly pesticides). The induction of beta-galactosidase activity was strictly dependent on the presence of seabream (Sparus aurata) betaa estrogen receptor (sbERbetaa) and substances known to have estrogenic activity. 17beta-Estradiol (E(2)) and diethylstilbestrol (DES), both agonists, were most active and the antagonist tamoxifen (TAM) was 14-fold less active than E(2). Among the chemicals tested bisphenol-A was most active, followed by pentachlorophenol and naphthalene. Ligand-binding assays with recombinant sbERbetaa and sbERalpha revealed that sbERbetaa binds E(2) with 6.5-fold higher affinity than sbERalpha, confirming the selection of a high sensitive receptor for the yeast assay. DES, ICI 182,780, estrone and TAM had higher relative binding affinity to E2 in sbERalpha than sbERbetaa, although there was no difference in IC50 for these steroids between the two receptors. These results reveal the usefulness of using the yeast-based receptor assay for detecting chemical interaction with steroid receptors from contaminated samples. PMID- 19303143 TI - Cellular accumulation and DNA platination of two new platinum(II) anticancer compounds based on anthracene derivatives as carrier ligands. AB - The anticancer properties of two new fluorescent platinum(II) compounds, cis [Pt(A9opy)Cl(2)] and cis-[Pt(A9pyp)(dmso)Cl(2)] are described. These compounds are highly active against several human tumor cell lines, including human ovarian carcinoma sensitive and cisplatin-resistant cell lines (A2780 and A2780R). To study the cellular processing of these new compounds, a series of in vitro studies have been performed, including the investigation of intracellular platinum accumulation and DNA-platination experiments in A2780 and A2780R cells. Compared to cisplatin, both compounds are accumulated highly in both sensitive and resistant cell lines, and more platinum has been found to bind to the nuclear DNA. Interestingly, cis-[Pt(A9opy)Cl(2)] shows high accumulation and DNA adduct formation in the resistant cell line A2780R, as compared to the sensitive counterpart A2780 cell line. This suggests that cis-[Pt(A9opy)Cl(2)] is able to overcome some of the well-known resistance mechanisms in this cell line, such as decreased cellular uptake and increased DNA repair. PMID- 19303144 TI - Synthesis, characterization and electrocatalysis of diiron propanediselenolate derivatives as the active site models of [FeFe]-hydrogenases. AB - As an extension of our study on the H-cluster model compounds, a series of diiron propanediselenolate (PDS)-type models have been successfully synthesized. Reaction of diselenol HSe(CH(2))(3)SeH with Fe(3)(CO)(12) in THF (tetrahydrofuran) at reflux gave the parent model compound [micro-Se(CH(2))(3)Se micro]Fe(2)(CO)(6) (1) in 48% yield. Further reaction of 1 with PPh(3) or PPh(2)H in the presence of Me(3)NO in MeCN at room temperature afforded the phosphine monosubstituted model compounds [micro-Se(CH(2))(3)Se-micro]Fe(2)(CO)(5)(L) (2, L=PPh(3); 3, L=PPh(2)H) in 76% and 68% yields, respectively. Similarly, the N heterocyclic carbene I(Mes)-monosubstituted model compound [micro-Se(CH(2))(3)Se micro]Fe(2)(CO)(5)(I(Mes)) (4) could be prepared in 46% yield by reaction of imidazolium salt I(Mes).HCl with n-BuLi followed by treatment of the resulting I(Mes) ligand with 1 in THF at room temperature. Compounds 1-4 were fully characterized by elemental analysis and various spectroscopic methods. While the structures of 1-4 were further confirmed by X-ray crystallography, the comparative study of 1 and its analog [micro-S(CH(2))(3)S-micro]Fe(2)(CO)(6) demonstrates that 1 is a better catalyst for TsOH proton reduction to hydrogen under electrochemical conditions. PMID- 19303145 TI - Difference in prevalence of common mental disorder as measured using four questionnaire delivery methods among young people in rural Zimbabwe. AB - BACKGROUND: Previous studies have suggested that interviewer-administered questionnaires can under-estimate the prevalence of depression and suicidal ideation when compared with self-administered ones. We report here on differences in prevalence of reporting mental health between four questionnaire delivery modes (QDM). METHODS: Mental health was assessed using the Shona Symptom Questionnaire (SSQ), a locally validated 14-item indigenous measure for common mental affective disorders. A representative sample of 1495 rural Zimbabwean adolescents (median age 18) was randomly allocated to one of four questionnaire delivery modes: self-administered questionnaire (SAQ), SAQ with audio (AASI), interviewer-administered questionnaire (IAQ), and audio computer-assisted survey instrument (ACASI). RESULTS: Prevalence of common affective disorders varied between QDM (52.3%, 48.6%, 41.5%, and 63.6% for SAQ, AASI, IAQ, and ACASI respectively (P<0.001)). Fewer participants failed to complete SSQ using IAQ and ACASI than other methods (1.6% vs. 12.3%; P<0.001). Qualitative data suggested that respondents found it difficult answering questions honestly in front of an interviewer. LIMITATIONS: Direction of accuracy cannot be ascertained due to lack of objective or clinical assessments of affective disorders. CONCLUSIONS: Estimates of prevalence of psychosomatic symptoms and suicidal ideation varied according to mode of interview. As each mode's direction of accuracy remains unresolved evaluations of interventions continue to be hampered. PMID- 19303146 TI - Middle ear adenoma with neuroendocrine differentiation (MEA-ND) in the pediatric population. AB - Middle ear adenoma with neuroendocrine differentiation is an uncommon tumor of the tympanic cavity. The terminology of this entity has suffered due to a poor understanding of the differentiation of these neoplasms, and has included both "adenoma" as well as "carcinoid tumor." Immunohistochemical techniques have helped clarify that these tumors all share elements of neuroendocrine differentiation. Occurrences of this tumor in the pediatric population are extremely rare. In the world literature, only three pediatric cases of middle ear adenoma with neuroendocrine differentiation have been described. We report the youngest case to date of middle ear adenoma with neuroendocrine differentiation, in a 13-year-old boy. We discuss the differential diagnosis, surgical management, and pathology of this entity. PMID- 19303147 TI - Differential expression of toll-like receptors 2 and 4 in rat middle ear. AB - OBJECTIVE: The epithelial cells of the middle ear and Eustachian tube must maintain an adequate mucosal defense system against various antigenic stimuli. Since toll-like receptors (TLRs) are known to play a critical role in mucosal defense, we investigated their expression in the mucosa of the tubotympanum, nasopharynx, and oral cavity of the rat. METHOD: The expression of TLR2 and TLR4 was examined in the mucosa of the tubotympanum, nasopharynx and oral cavity of the rat using real time RT-PCR and Western blot analysis. RESULTS: Transcripts for TLR2 and TLR4 were detected in the mucosa of the tubotympanum, nasopharynx, and oral cavity of the rat. The expression of TLR2 and TLR4 in the middle ear was increased more than in the other anatomical areas. Differential expression of these molecules at the protein level was confirmed by Western blot analysis. CONCLUSIONS: Diverse expression of TLR2 and TLR4 in different parts of the tubotympanum and upper aerodigestive tract suggests region-specific functional modulation of the innate immune system. Differential expression of subtypes of the TLR in the normal physiology of the tubotympanum and upper aerodigestive tract also suggests that they may play a role in the pathophysiology of otitis media. PMID- 19303148 TI - Bilateral cochlear implantation in children with Noonan syndrome. AB - Noonan syndrome is a mostly autosomal dominant inherited disorder, which can be accompanied by hearing disorders or deafness, coagulation disorders, combined heart defects and developmental disorders. We are reporting on two children with an established Noonan syndrome with a severe bilateral hearing loss of respectively 95 and 100 dB and proper findings in the CT/MRI of the petrous bone. After complete otologic and radiologic diagnostics, both children underwent bilateral cochlear implantation successfully. According to the authors' knowledge, this is the first time that cochlear implant therapy is discussed in patients with Noonan syndrome. PMID- 19303149 TI - Acute otitis media due to Haemophilus influenzae with antibiotic resistance: experience of a tertiary hospital in Tokyo city. AB - OBJECTIVE: The objective of the study was to clarify the associated factors of pediatric acute otitis media (AOM) due to Haemophilus influenzae with antibiotic resistance. METHODS: A retrospective chart review was conducted for all AOM cases due to H.influenzae in children younger than 2 years of age that were presented at the Department of Otolaryngology, Japanese Red Cross Medical Hospital (JRCMC), Tokyo, from April 2005 to March 2007. RESULTS: Seventy-six children were enrolled in this study. Multivariate analysis showed that residing at the JRCMC infant home (OR, 3.7; [95% CI, 1.1-12.8]; P=0.039) and a history of consuming macrolide in the past month (OR, 4.7; [95% CI, 1.1-21.5]; P=0.041) were the associated factors for AOM due to beta-lactamase positive, ampicillin-resistant H. influenzae. CONCLUSIONS: A history of consuming macrolide in the past month should be noted on antibiotic prescriptions for children with AOM who are younger than 2 years of age. PMID- 19303150 TI - Preparation, characterization of NIPAM and NIPAM/BAM copolymer nanoparticles and their acute toxicity testing using an aquatic test battery. AB - Poly N-isopropylacrylamide and N-isopropylacrylamide/N-tert-butylacrylamide copolymer nanoparticles of 50-70 nm were prepared by free radical polymerisation. The particle sizes of the copolymer nanoparticles were measured in the test media Milli-Q water, Algae Media, Daphnia Media and Microtox Diluent as a function of temperature. Whereas in Milli-Q water the particle size was seen to decrease above the lower critical solution temperature of the thermoresponsive polymer, in the test media it was seen to increase significantly, indicative of aggregation. At the temperatures employed for the ecotoxicological studies all particles, with the exception of the 50:50 copolymer existed as nanoparticles. The zeta potential of Poly N-isopropylacrylamide and N-isopropylacrylamide/N-tert-butylacrylamide copolymer particles measured in the different media was seen to correlate well with the ratio of N-tert-butylacrylamide monomer and therefore the hydrophobicity of the particles. Ecotoxicological studies of the copolymer nanoparticles was performed using four test species Vibrio fischeri, Pseudokirchneriella subcapitata, Daphnia magna, Thamnocephalus platyurus and the cytotoxicity of the 100% Poly N-isopropylacrylamide and 85:15 N-isopropylacrylamide/N-tert butylacrylamide copolymer nanoparticles was evaluated using a salmonid cell line. Although no significant cytotoxicological response was recorded, significant ecotoxicological response was observed at particle concentrations of up to 1000 mg l(-1). The ecotoxicological response was seen to correlate well with the ratio of N-tert-butylacrylamide monomer and therefore with the zeta potential of the nanoparticles. The toxic response in Daphnia magna was seen to further correlate with the reduction in zeta potential pointing towards a contribution of secondary effects due to modification of the medium. No correlation with particle size was observed. The sensitivity of the test species was seen to vary depending on copolymer composition. The relevance of the derived structure-activity relationships is discussed. PMID- 19303151 TI - Description and repeatability of a newly developed spinal cord injury scale for dogs. AB - The objectives of this study were to describe a new spinal cord injury scale for dogs, evaluate repeatability through determining inter-rater variability of scores, compare these scores to another established system (a modified Frankel scale), and determine if the modified Frankel scale and the newly developed scale were useful as prognostic indicators for return to ambulation. A group of client owned dogs with spinal cord injury were examined by 2 independent observers who applied the new Texas Spinal Cord Injury Score (TSCIS) and a modified Frankel scale that has been used previously. The newly developed scale was designed to describe gait, postural reactions and nociception in each limb. Weighted kappa statistics were utilized to determine inter-rater variability for the modified Frankel scale and individual components of the TSCIS. Comparisons were made between raters for the overall TSCIS score and between scales using Spearman's rho. An additional group of dogs with surgically treated thoracolumbar disk herniation was enrolled to look at correlation of both scores with spinal cord signal characteristics on magnetic resonance imaging (MRI) and ambulatory outcome at discharge. The actual agreement between raters for the modified Frankel scale was 88%, with a weighted kappa value of 0.93. The TSCIS had weighted kappa scores for gait, proprioceptive positioning and nociception components that ranged from 0.72 to 0.94. Correlation between raters for the overall TSCIS score was Spearman's rho=0.99 (P<0.001). Comparison of the overall TSCIS score to the modified Frankel score resulted in a Spearman's rho value of 0.90 (P<0.001). The modified Frankel score was weakly correlated with the length of hyperintensity of the spinal cord: L2 vertebral body length ratio on mid-sagittal T2-weighted MRI (Spearman's rho=-0.45, P=0.042) as was the overall TSCIS score (Spearman's rho= 0.47, P=0.037). There was also a significant difference in admitting modified Frankel scores (P=0.029) and admitting overall TSCIS scores (P=0.02) between dogs that were ambulatory at discharge and those that were not. Results from this study suggest that the TSCIS is an easy to administer scale for evaluating canine spinal cord injury based on the standard neurological exam and correlates well with a previously described modified Frankel scale. PMID- 19303152 TI - Danish dairy farmers' perceptions and attitudes related to calf-management in situations of high versus no calf mortality. AB - Calf mortality has remained at a high (risk approximately 7%) in Denmark, despite much knowledge disseminated to farmers. We conducted a qualitative interview to explore calf managers' perception of their own management and working situation, and how this related to calf mortality problems in their herds. Twenty-eight calf managers from herds with either no (NO) or high (HI; 17-31%) calf mortality in 2004 were included in the study. We used our judgement to create a model of the inter-relationships among the attitudes that seemed most different between the two groups of farmers. 'Flexible time,' (for activities such as management of crisis or unexpected events) was crucial; it allowed the farmer to intervene immediately at first sign of a crisis. Our interpretation suggests that focus on the farmers time planning and structure of everyday activities is, therefore, a key to understanding and solving problems related to calf disease and mortality. An important distinction between the groups was whether the calf manager had a basic belief that calf mortality is a permanent crisis that has to be expected to be present on a dairy farm. Previous experience in solving disease and mortality problems in calves had a strong positive influence on this belief. We suggest that the issues of empowerment and flexible time management be explored further to assist dairy-calf raisers. PMID- 19303153 TI - Evidence from the UK Zoonoses Action Plan in favour of localised anomalies of Salmonella infection on United Kingdom pig farms. AB - Salmonella spp. are important food-borne pathogens. Abattoir studies demonstrated that almost a quarter of British finisher pigs might carry Salmonella, which led to the introduction by the British Pig Executive of their Zoonoses Action Plan (ZAP) to monitor the Salmonella status of United Kingdom pig farms by testing meat juice samples using an ELISA system. We used the K-function and approaches from the field of geostatistics to study routine data from ZAP. We demonstrated that there is statistical evidence that geographically localized anomalies of Salmonella infection were present in one of three regions studied. The physical mechanisms underlying this structure remain unclear: spatial structure might be present as a result of shared spatially structured (second-order) or non spatially structured (first-order) risk factors, transmission processes, or a combination of both. We have demonstrated a way to use routinely collected surveillance data to enhance the knowledge of spatial disease epidemiology. PMID- 19303154 TI - Veterinarian injuries associated with bovine TB testing livestock in Michigan, 2001. AB - Testing all the cattle in an entire state with a uniform procedure for each animal affords an opportunity to relate human injury data to a known number of animals handled while carrying out a standardized procedure. Our objective was to describe the type and incidence density of injuries associated with TB-testing a large number of cattle herds, and to delineate the various factors associated with the risk of injury. A survey was mailed to all veterinarians (N = 259) who had completed at least five official bovine TB (bTB) herd tests in Michigan in 2001. We collected data regarding basic demographics and health status, work experience, veterinary specialty, and practice information. Each veterinarian was also requested to complete a separate injury questionnaire for each injury received while TB testing livestock in 2001. Accurate addresses were found for 247 eligible veterinarians, 175 (71%) of whom returned the survey. Thirty-six veterinarians reported a total of 53 injuries (10 major, 12 minor and 31 self treated). Hands (29%) and legs (21%) were the anatomic locations most frequently injured, with sprains/strains (30%) and abrasion/contusion (30%) the most common types of injuries sustained. The overall incidence density of injuries was 1.9 per 10,000 animals tested. Female gender (RR = 3.3), being employed by the government (RR = 4.5), and smoking (RR = 6.0) were significantly associated with a higher rate of injury. Significant colliniearities were found between some risk factors associated with an increased rate of injury and participants thought 81% of their injuries could have been prevented. These results are explained by the administrative structure of the bTB testing program in Michigan, and the changing demographics of the veterinary workforce. PMID- 19303155 TI - An object-oriented Bayesian network modeling the causes of leg disorders in finisher herds. AB - The implementation of an effective control strategy against disease in a finisher herd requires knowledge regarding the disease level in the herd. A Bayesian network was constructed that can estimate risk indexes for three cause-categories of leg disorders in a finisher herd. The cause-categories of leg disorders were divided into infectious causes (arthritis caused by infectious pathogens), physical causes (e.g. fracture and claw lesions), and inherited causes (osteochondrosis). Information about the herd (e.g. the herd size, floor type and number of suppliers) and information about individual pigs (e.g. results from diagnostic tests) were used to estimate the most likely cause of leg disorders at herd level. As information to the model originated from two different levels, we used an object-oriented structure in order to ease the specification of the Bayesian network. Hence, a Herd class and a Pig class comprised the basic components of the object-oriented structure. The causal structure of the model was based on evidence from published literature. The conditional probabilities used in the model were elicited from experts within the field and from the published literature. To illustrate the behaviour of the model, we investigated the value of different levels of evidence in two fictitious herds with different herd characteristics related to the risk of leg disorders (e.g. purchase policy, production type and the stocking density in pens). The model enabled us to demonstrate the value of performing systematic collection of additional information (i.e. clinical, pathological and bacteriological examination) when identifying causes of leg disorders at herd level. PMID- 19303156 TI - Hepatitis C virus NS5A and core proteins induce oxidative stress-mediated calcium signalling alterations in hepatocytes. AB - BACKGROUND/AIMS: The hepatitis C virus (HCV) structural core and non-structural NS5A proteins induce in liver cells a series of intracellular events, including elevation of reactive oxygen and nitrogen species (ROS/RNS). Since oxidative stress is associated to altered intracellular Ca(2+) homeostasis, we aimed to investigate the effect of these proteins on Ca(2+) mobilization in human hepatocyte-derived transfected cells, and the protective effect of quercetin treatment. METHODS: Ca(2+) mobilization and actin reorganization were determined by spectrofluorimetry. Production of ROS/RNS was determined by flow cytometry. RESULTS: Cells transfected with NS5A and core proteins showed enhanced ROS/RNS production and resting cytosolic Ca(2+) concentration, and reduced Ca(2+) concentration into the stores. Phenylephrine-evoked Ca(2+) release, Ca(2+) entry and extrusion by the plasma membrane Ca(2+)-ATPase were significantly reduced in transfected cells. Similar effects were observed in cytokine-activated cells. Phenylephrine-evoked actin reorganization was reduced in the presence of core and NS5A proteins. These effects were significantly prevented by quercetin. Altered Ca(2+) mobilization and increased calpain activation were observed in replicon containing cells. CONCLUSIONS: NS5A and core proteins induce oxidative stress mediated Ca(2+) homeostasis alterations in human hepatocyte-derived cells, which might underlie the effects of both proteins in the pathogenesis of liver disorders associated to HCV infection. PMID- 19303157 TI - Variable activation of phosphoinositide 3-kinase influences the response of liver grafts to ischemic preconditioning. AB - BACKGROUND/AIMS: The efficacy of ischemic preconditioning (IPC) in preventing reperfusion injury in human liver transplants is still questioned. Phosphoinositide-3-kinase (PI3K) is essential for IPC development in rodent livers. This work investigates whether PI3K-dependent signals might account for the inconsistent responses to IPC of transplanted human livers. METHODS: Forty livers from deceased donors were randomized to receive or not IPC before recovery. PI3K activation was evaluated in biopsies obtained immediately before IPC and 2 h after reperfusion by measuring the phosphorylation of the PI3K downstream kinase PKB/Akt and the levels of the PI3K antagonist phosphatase tensin-homologue deleted from chromosome 10 (PTEN). RESULTS: IPC increased PKB/Akt phosphorylation (p = 0.01) and decreased PTEN levels (p = 0.03) in grafts, but did not significantly ameliorate post-transplant reperfusion injury. By calculating T(2h)/T(0) PKB/Akt phosphorylation ratios, 10/19 (53%) of the preconditioned grafts had ratios above the control threshold (IPC-responsive), while the remaining nine grafts showed ratios comparable to controls (IPC-non responsive). T(2h)/T(0) PTEN ratios were also decreased (p < or = 0.03) only in IPC-responsive grafts. The patients receiving IPC-responsive organs had ameliorated (p < or = 0.05) post-transplant aminotransferase and bilirubin levels, while prothrombin activity was unchanged. CONCLUSIONS: Impaired PI3K signaling might account for the variability in the responses to IPC of human grafts from deceased donors. PMID- 19303158 TI - HCV replication suppresses cellular glucose uptake through down-regulation of cell surface expression of glucose transporters. AB - BACKGROUND/AIMS: Persistent infection with hepatitis C virus (HCV) causes extrahepatic diseases, including diabetes. We investigated the possible effect(s) of HCV replication on cellular glucose uptake and expression of the facilitative glucose transporter (GLUT) 2 and 1. METHODS: We used Huh-7.5 cells harboring either an HCV subgenomic RNA replicon (SGR) or an HCV full-genomic RNA replicon (FGR), HCV-infected cells, and the respective cells treated with interferon (IFN). We also used liver tissue samples obtained from patients with or without HCV infection. RESULTS: Glucose uptake and surface expression of GLUT2 and GLUT1 were suppressed in SGR, FGR and HCV-infected cells compared to the control cells. Expression levels of GLUT2 mRNA, but not GLUT1 mRNA, were lower in SGR, FGR and HCV-infected cells than in the control. Luciferase reporter assay demonstrated decreased GLUT2 promoter activities in SGR, FGR and HCV-infected cells. IFN treatment restored glucose uptake, GLUT2 surface expression, GLUT2 mRNA expression and GLUT2 promoter activities. Also, GLUT2 expression was reduced in hepatocytes of liver tissues obtained from HCV-infected patients. CONCLUSIONS: HCV replication down-regulates cell surface expression of GLUT2 partly at the transcriptional level, and possibly at the intracellular trafficking level as suggested for GLUT1, thereby lowering glucose uptake by hepatocytes. PMID- 19303159 TI - Elevated expression of DKK1 is associated with cytoplasmic/nuclear beta-catenin accumulation and poor prognosis in hepatocellular carcinomas. AB - BACKGROUND/AIMS: To assess the value of Dickkopf-1 (DKK1) for predicting clinical outcome of human hepatocellular carcinoma (HCC) in patients with HCC. METHODS: Expression of DKK1 and beta-catenin was investigated in HCC cell lines using qRT PCR, Western blotting and immunofluorescence. Tissue microarrays representing 314 HCC patients were used to determine the expression patterns of DKK1 and beta catenin by immunohistochemistry, and prognostic significance was assessed by using Kaplan-Meier survival estimates and log-rank tests. RESULTS: The expression level of DKK1 was associated with the staining pattern of beta-catenin in HCC cell lines, and DKK1 overexpression correlated with beta-catenin cytoplasmic/nuclear accumulation in clinical HCC samples (P=0.011, correlation coefficient=0.144). High DKK1 expression predicted unfavorable prognosis in HCC patients, especially in early stage patients and those with normal AFP levels. In multivariate analyses, DKK1 was an independent predictor for overall survival (OS) (P=0.002) and disease-free survival (DFS) (P=0.002) of HCC patients. Furthermore, the HCC patients with high DKK1 expression and cytoplasmic/nuclear beta-catenin accumulation had very poor prognosis. CONCLUSIONS: Elevated expression of DKK1 is a critical event in patients with HCC that indicates poor clinical outcome. DKK1, alone or combined with beta-catenin, is a novel prognostic predictor for HCC patients. PMID- 19303160 TI - Heparanase inhibitor PI-88 as adjuvant therapy for hepatocellular carcinoma after curative resection: a randomized phase II trial for safety and optimal dosage. AB - BACKGROUND/AIMS: Hepatocellular carcinoma recurrence after curative treatment adversely influences clinical outcome. It is important to explore adjuvant therapies. This phase II/stage 1 multi-center, randomized trial investigated the safety, optimal dosage and preliminary efficacy of PI-88, a novel heparanase inhibitor, in the setting of post-operative recurrence of HCC according to a Simon's 2-stage design. METHODS: Three groups were included: one untreated arm (Group A) and two PI-88 arms (Group B: 160 mg/day; Group C: 250 mg/day). Treatment groups received PI-88 over nine 4-week treatment cycles, followed by a 12-week treatment-free period. Safety and optimal dosage were assessed. RESULTS: Overall, 172 patients were randomized and 168 were included in the intention-to treat (ITT) population. Treatment-related adverse effects included cytopenia, injection site hemorrhage, PT prolongation, etc. Four serious adverse events were possibly related to PI-88 treatment. One (1.8%) group B patients and six (10.5%) group C had hepatotoxicity-related withdrawals. Among the ITT population, 29 patients (50%) in Group A, 35 (63%) in Group B, and 22 (41%) in Group C remained recurrence-free at completion. Calculated T(1) value suggested 160 mg/day treatment satisfied the criteria for the next stage of the trial. CONCLUSIONS: PI 88 at 160 mg/day is optimal and safe, and shows preliminary efficacy as an adjunct therapy for post-operative HCC. PMID- 19303161 TI - Genetic factors contribute to variation in serum alanine aminotransferase activity independent of obesity and alcohol: a study in monozygotic and dizygotic twins. AB - BACKGROUND/AIMS: This study aimed to determine the heritability of serum alanine aminotransferase (S-ALT) and fasting serum insulin (fS-insulin) concentration as well as determine the association of these measures with liver fat content in young adult monozygotic (MZ) and dizygotic (DZ) twins. METHODS: Three hundred and thirteen individual twins were recruited from a population-based cohort (n = 4929). The study subjects represented a wide range of body mass indexes (BMI), were free of any diseases or regular medications and had an intake of less than two drinks of alcohol/day. To verify that S-ALT is a marker of liver fat, it was measured by proton magnetic resonance spectroscopy ((1)H MRS) in 66 subjects. Heritability estimations were performed using BMI- and gender-adjusted values. RESULTS: Intra-pair correlations were significantly higher in the MZ twins than the DZ twins for both S-ALT (0.65 for MZ and 0.04 for DZ) and fS-insulin (0.58 and 0.34, respectively). Heritability of S-ALT was 55% and that of fS-insulin 61%. In the 66 subjects S-ALT (r = 0.70 for women and r = 0.50 for men, p < or = 0.01 for both) and fS-insulin (r = 0.58 and r = 0.59, respectively, p < or = 0.01 for both) concentrations correlated significantly with liver fat content. CONCLUSIONS: These twin data suggest that approximately 60% of the variation in S ALT, a marker of liver fat content, is genetically determined. PMID- 19303162 TI - Prevention of hepatocellular carcinoma: the holy grail of hepatitis B treatment. PMID- 19303163 TI - Hepatic venous pressure gradient predicts development of hepatocellular carcinoma independently of severity of cirrhosis. AB - BACKGROUND/AIMS: A total of 213 patients with compensated cirrhosis, portal hypertension and no varices were included in a trial evaluating beta-blockers in preventing varices. Predictors of the development of hepatocellular carcinoma (HCC), including hepatic venous pressure gradient (HVPG) were analyzed. METHODS: Baseline laboratory tests, ultrasound and HVPG measurements were performed. Patients were followed prospectively every three months until development of varices or variceal bleeding or end of the study in 09/02. The endpoint was HCC development according to standard diagnostic criteria. Univariate and multivariate Cox regression models were developed to identify predictors of HCC. RESULTS: In a median follow-up of 58 months 26/213 (12.2%) patients developed HCC. Eight patients were transplanted and 28 patients died without HCC. Twenty one (84%) HCC developed in patients with HCV. On multivariate analysis HVPG (HR 1.18; 95%CI 1.08-1.29), albumin (HR 0.34; 95%CI 0.14-0.83) and viral etiology (HR 4.59; 95%CI 1.51-13.92) were independent predictors of HCC development. ROC curves identified 10 mmHg of HVPG as the best cut-off; those who had an HVPG above this value had a 6-fold increase in the HCC incidence. CONCLUSIONS: Portal hypertension is an independent predictor of HCC development. An HVPG >10 mmHg is associated with a 6-fold increase of HCC risk. PMID- 19303164 TI - Prospects for retinal gene replacement therapy. AB - Inherited retinal degeneration, which includes conditions such as retinitis pigmentosa and Leber congenital amaurosis (LCA), affects approximately 1/3000 of the population in the Western world. It is characterized by loss of vision and results from mutations in any one of >100 different genes. There are currently no effective treatments, but many of the genes have now been identified and their functions elucidated, providing a major impetus to develop gene-based treatments. Preliminary results from three clinical trials indicate that the treatment of a form of LCA by gene therapy can be safe and effective. Here, we discuss the potential for treating other forms of retinal degeneration by gene therapy, focusing on the gene defects that are likely to be the most amenable to treatment. PMID- 19303165 TI - Plastid signalling to the nucleus: messengers still lost in the mists? AB - The concept of plastid signalling posits that signals originating from chloroplasts modulate nuclear gene expression (NGE). Put simply, it claims that signalling factors are exported from the chloroplast, traverse the cytosol, and act in the nucleus. Pertinent signals are thought to derive from various sources, including the tetrapyrrole pathway, protein synthesis, reactive oxygen species, or the redox state of the organelle. Recent studies have cast doubt on the most popular candidate signalling molecule, the tetrapyrrole pathway intermediate Mg protoporphyrin IX, indicating that chloroplast activity might control NGE indirectly by affecting cytosolic metabolite levels or redox states (metabolic signalling). Here, we focus on recent developments and confusions in the field of plastid signalling research and highlight alternative scenarios of plastid nucleus signal transduction. Future analyses of chloroplast-nucleus communication should focus on providing an integrated view of plastid signalling under physiologically relevant conditions. PMID- 19303166 TI - Human olfaction: from genomic variation to phenotypic diversity. AB - The sense of smell is a complex molecular device, encompassing several hundred olfactory receptor proteins (ORs). These receptors, encoded by the largest human gene superfamily, integrate odorant signals into an accurate 'odor image' in the brain. Widespread phenotypic diversity in human olfaction is, in part, attributable to prevalent genetic variation in OR genes, owing to copy number variation, deletion alleles and deleterious single nucleotide polymorphisms. The development of new genomic tools, including next generation sequencing and CNV assays, provides opportunities to characterize the genetic variations of this system. The advent of large-scale functional screens of expressed ORs, combined with genetic association studies, has the potential to link variations in ORs to human chemosensory phenotypes. This promises to provide a genome-wide view of human olfaction, resulting in a deeper understanding of personalized odor coding, with the potential to decipher flavor and fragrance preferences. PMID- 19303167 TI - A statistical metadata model for clinical trials' data management. AB - We introduce a statistical, process-oriented metadata model to describe the process of medical research data collection, management, results analysis and dissemination. Our approach explicitly provides a structure for pieces of information used in Clinical Study Data Management Systems, enabling a more active role for any associated metadata. Using the object-oriented paradigm, we describe the classes of our model that participate during the design of a clinical trial and the subsequent collection and management of the relevant data. The advantage of our approach is that we focus on presenting the structural inter relation of these classes when used during datasets manipulation by proposing certain transformations that model the simultaneous processing of both data and metadata. Our solution reduces the possibility of human errors and allows for the tracking of all changes made during datasets lifecycle. The explicit modeling of processing steps improves data quality and assists in the problem of handling data collected in different clinical trials. The case study illustrates the applicability of the proposed framework demonstrating conceptually the simultaneous handling of datasets collected during two randomized clinical studies. Finally, we provide the main considerations for implementing the proposed framework into a modern Metadata-enabled Information System. PMID- 19303168 TI - A pilot study examining undesirable events among emergency department-boarded patients awaiting inpatient beds. AB - STUDY OBJECTIVE: We describe the frequency of undesirable events among patients boarding at a single, urban, tertiary, teaching emergency department (ED) through retrospective chart abstraction. METHODS: This was a chart review of all patients admitted during 3 randomly selected days in 2003 (n=162) to track the frequency of undesirable events such as missed relevant home medications, missed laboratory test results, arrhythmias, or other adverse events. RESULTS: One hundred fifty one charts were abstracted (93.2%); 27.8% had an undesirable event, 17.9% missed a relevant home medication, and 3.3% had a preventable adverse event. There was a higher frequency of undesirable events among older patients (35.9%, aged >50 years; 7.3%, aged 20 to 49 years; 28.6%, aged 0 to 19 years) and those with more comorbidities (44.4% among Charlson score >or=3; 30.8% score 2; 36.1% score 1; 14.5% score 0). CONCLUSION: A substantial frequency of undesirable events occurs while patients board in the ED. These events are more frequent in older patients or those with more comorbidities. Future studies need to compare the rates of undesirable events among patients boarding in the ED versus inpatient units. PMID- 19303169 TI - [Three years with "Tobacco-control law": cleaner air but not clean enough]. PMID- 19303170 TI - [Reference values of the Spanish version of the SF-12v2 for the diabetic population]. AB - OBJECTIVE: Perceived health status questionnaires provide important information for health outcomes research. Reference measures are required to interpret the health-related quality of life questionnaires belonging to the short form (SF) health survey family. Previous studies have provided these reference measures for the Spanish general population, but not for specific disease conditions. The aim of the present study was to obtain diabetic population-based norms for the Spanish version of the 12-item short form health survey version II (SF-12v2) in the region of Murcia (Spain). METHODS: We performed a cross-sectional telephone survey in 1,500 non-institutionalized patients with type 1 or 2 diabetes, aged 18 years or older and representative of the region of Murcia. The central position, dispersion statistics and percentiles were calculated for each of the eight SF 12v2 scales and summary components (physical and mental). RESULTS: The mental component summary (MCS) (mean: 50.5+/-12.8) was higher than the physical component summary (PCS) (42.5+/-11.8). By sex, men had more favorable MCS (53.6+/ 11.6) and PCS (44.9+/-11.2) scores than women (MCS: 47.7+/-13.2; PCS: 40.3+/ 11.9). The dimension with the highest score was vitality (men: 57.2+/-11.1; women: 51.9+/-12.1), while that with the lowest score was general health (men: 39.0+/-9.7; women: 35.8+/-10.0). When the groups were stratified by age, scores continued to be higher in men than in women. CONCLUSIONS: The results of the present study should be taken as the diabetic population-based norms for the Spanish version of the SF-12v2 in the region of Murcia. These results may be useful to establish therapeutics targets, as well as for comparisons with the general, healthy and ill populations. PMID- 19303171 TI - Novel ureas and thioureas of 15-membered azalides with antibacterial activity against key respiratory pathogens. AB - The new ureas and thioureas of 15-membered azalides, N''-substituted 9a-(N' carbamoyl-gamma-aminopropyl) (4), 9a-(N'-thiocarbamoyl-gamma-aminopropyl) (6), 9a [N'-(beta-cyanoethyl)-N'-(carbamoyl-gamma-aminopropyl)] (8) and 9a-[N'-(beta cyanoethyl)-N'-(thiocarbamoyl-gamma-aminopropyl)] (10) of 9-deoxo-9-dihydro-9a aza-9a-homoerythromycin A (2), were synthesized and structurally characterized by NMR and IR spectroscopic methods and mass spectrometry. The new compounds were evaluated in vitro against a panel of erythromycin susceptible and erythromycin resistant gram-positive and gram-negative bacterial strains. These compounds displayed an excellent overall antibacterial in vitro activity against erythromycin sensitive gram-positive strains, Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, and good against negative strains, Moraxella catarrhalis and Haemophilus influenzae. In addition, several ureas with naphthyl substituents (4f, 4g, 4h) showed better activity in comparison to azithromycin against inducible resistant S. pyogenes. Ureas with naphthyl substituents 4g, 4h and thiourea 8h displayed moderate activity against constitutively resistant S. pneumoniae. PMID- 19303172 TI - 2D- and 3D-QSAR studies on 54 anti-tumor Rubiaceae-type cyclopeptides. AB - RA-VII, a bicyclic hexapeptide isolated from the roots of Rubia cordifolia, Rubia akane belongs to Rubiaceae-type cyclopeptides (RAs) and has attracted much attention for its potent anti-tumor activity and its bicyclic structure incorporating the isodityrosine moiety. In this work, hologram quantitative structure-activity relationship (HQSAR), comparative molecular field analysis (CoMFA), and comparative molecular similarity indices analysis (CoMSIA) methods were employed to develop 2D- and 3D-QSAR models for 54 anti-tumor RAs. The LOO cross-validated q(2) values of HQSAR, CoMFA and CoMSIA models are 0.701, 0.510 and 0.613, respectively. The predictive ability of these models was validated by the test set including 7 RAs, and the predicted IC(50) values were in good agreement with the experimental IC(50) values. HQSAR result showed that chirality descriptor plays an important role in anti-tumor activity of RAs and OMe at R(1) and R(2) is necessary for increasing their activity. CoMFA and CoMSIA results demonstrated that small bulky and electropositive side chains at R(3) position and hydrophobic groups at R(7) and R(8) positions will increase their activity, and intra-molecular hydrogen bonds between residues 1 and 4 are necessary to maintain the pharmacophoric conformation of RAs. These results may be helpful in designing novel and potential anti-tumor RAs. PMID- 19303173 TI - A QSAR study on relationship between structure of sulfonamides and their carbonic anhydrase inhibitory activity using the eigenvalue (EVA) method. AB - In this study, we present an application of EVA descriptors for a QSAR model of inhibition of carbonic anhydrase isozyme CA II by an heterogeneous set of 66 sulfonamide compounds. For each of the compounds, geometry optimization and frequency calculations have been performed using the DFT/B3LYP level of the theory in conjugated with the 6-31G* basis set. Different numbers of EVA descriptors for each structure were produced by applying various values of Gaussian kernel of a fixed standard deviation, sigma (cm(-1)) and sampled at fixed increments of L (cm(-1)) during the evaluation of the descriptors based on their vibrational frequencies. The set of compounds was divided into two subsets. The first subset contained the 22 compounds that were used as the test compounds. The remaining 44 compounds were used as the training set. Several QSAR models have been developed using these calculated EVA descriptors and the carbonic anhydrase isozyme CA II inhibitory data (K(i)) of the compounds. Among the QSAR models evaluated, the one that produced the best statistical results had the parameters sigma and L both equal to 5 cm(-1). This model produced correlation values (R(2)) of 0.777 and 0.616 for the training and test sets, respectively. The results of this study showed that EVA descriptors perform well as explanatory and predictive tools for modeling the inhibition activity of carbonic anhydrase by a set of sulfonamide compounds. PMID- 19303174 TI - Synthesis and biochemical evaluation of a cyclic RGD oxorhenium complex as new ligand of alphaVbeta3 integrin. AB - We report the design of a new ligand of integrins that might be used for the molecular imaging of tumor neoangiogenesis. For this purpose, we designed a modified RGD tripeptide bearing a N-terminal N-bis(thioethyl)glycinate (NS(2)) motif and a thioethyl moiety at the C-terminus. Simultaneous coordination of an oxorhenium core by the NS(2) and thioethyl moieties led to peptide cyclization and gave the corresponding monomers 13a and b (major isomer) resulting from the syn/anti-isomerism, along with dimers' species 16a and b. Cyclometallated peptide 13b showed the most promising activity with an IC(50) of 86 nM for integrin alpha(V)beta(3) whereas it binds integrin alpha(IIb)beta(3) with an affinity lower by an order of magnitude. Labeling with [(99m)Tc]oxotechnetium gluconate led exclusively to complex 17, the equivalent of compound 13b, which displayed satisfactory stabilities in mice plasma and towards glutathione. PMID- 19303175 TI - [Cat scratch disease with bone involvement: a case report and literature review]. AB - INTRODUCTION: Cat scratch disease is an infectious disease caused by Bartonella henselae. Most of the patients present with a lymphadenopathy associated with a local infection at the site of the cat scratch. Disseminated infection is uncommon. CASE REPORT: We report an immunocompetent 61-year-old woman who presented with a systemic cat scratch disease including a multifocal osteomyelitis. Diagnosis was confirmed by PCR on the adenopathy. A literature review identified 51 other cases of osteomyelitis associated with cat scratch disease, 14 of those confirmed by PCR. CONCLUSION: Bone involvement in cat scratch disease is rare, especially in adults. The diagnosis should be suspected on the basis of patient questioning. The antibiotherapy and the place of surgery are discussed. PMID- 19303176 TI - [An unusual cause of bone pain in a young woman]. PMID- 19303177 TI - [Paroxysmal nocturnal hemoglobinuria]. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is a rare acquired disorder of hematopoietic stem cells. PNH is related to a somatic mutation in the phosphatidylinositol glycan class A (PIG-A), X-linked gene, responsible for a deficiency in glycosylphosphatidylinositol-anchored proteins (GPI-AP). The lack of one of the GPI-AP complement regulatory proteins (CD59) leads to haemolysis. The disease is diagnosed with haemolytic anemia, marrow failure or episodes of venous thrombosis. The diagnosis is based on flow cytometry, which allowed direct quantification of the GPI-AP-deficient cells. From earlier descriptions, the clinical polymorphism of PNH has been recognized by two presentations; one form, predominantly haemolytic without overt marrow failure, referred to classic PNH and the other one, with marrow failure, was often described as the aplastic anemia PNH syndrome (AA-PNH). Thromboses remain a major life threatening complication affecting outcomes in both disease subcategories. Thrombotic events are characterized by involvement of unusual sites (hepatic, mesenteric, cerebral, dermal veins). In classic PNH, recent studies have focused on inhibiting the complement cascade with encouraging clinical results using eculizumab, a C5 inhibitor humanized monoclonal antibody. Concerning the AA-PNH syndrome, bone marrow transplantation (BMT) is the reference treatment in young patients with a sibling donor. Immunosuppressive therapy remains an important treatment modality in this subcategory for patients without a donor or ineligible for BMT. Recurrent thrombotic events remains even now associated with bad prognosis, whatever the form of the disease. PMID- 19303178 TI - Tarsal strip technique for correction of malposition of the lower eyelid after treatment of orbital trauma. AB - PURPOSE: The transconjunctival, subciliary, subtarsal, and subpalpebral approaches for accessing the infraorbital rim and orbital floor have both advantages and disadvantages. The most common complications include rounding of the lateral canthal angle, lower eyelid retraction with inferior scleral show, and frank ectropion. MATERIALS AND PATIENTS: From 2000 to 2007, we treated 29 patients with lower eyelid malposition after surgery to manage the floor and infraorbital trauma (22 subciliary approaches, five transconjunctival approaches and lateral canthotomies, and two transconjunctival approaches). To correct lower eyelid malposition, we applied the tarsal strip technique in all patients. RESULTS: Twenty-five patients had scleral show and four patients had ectropion: three were previous treated using transconjunctival access and one using subciliary access. Twenty-six patients obtained satisfactory correction of eyelid malposition in a single-step surgical procedure, while three patients required a second surgical step to correct the remaining scleral show. Good aesthetic and functional results were achieved in all cases. CONCLUSIONS: All approaches to the infraorbital rim or orbital floor have the potential for postoperative sequelae. The tarsal strip technique is a relatively simple technique that oral and maxillofacial surgeons can use to manage lower lid malposition, such as scleral show and ectropion. PMID- 19303179 TI - Co-activation differences in lower limb muscles between asymptomatic controls and those with varying degrees of knee osteoarthritis during walking. AB - BACKGROUND: Increased muscle co-activation during gait has been identified as a neuromuscular alteration associated with knee osteoarthritis, however levels of co-activation among different osteoarthritis severity have not been established. The purpose of this study was to determine if differences in co-activation could be detected among asymptomatic controls, those with moderate and those with severe osteoarthritis using a co-activation index and a pattern recognition technique. METHODS: Surface electromyograms from vastus lateralis and medialis, lateral and medial hamstring and gastrocnemius pairs were recorded from 63 asymptomatic, 59 moderate and 48 severe osteoarthritic subjects during self selected walking. A co-activation index was calculated over the initial stance for four medial and lateral muscle pairs. The four co-activation indices were tested among groups using a one factor ANOVA (alpha=0.05). Gait waveform pattern recognition procedures were applied to yield a principal pattern, scored for each muscle site and subject. A mixed model ANOVA (group-muscle) tested for principal pattern score differences. FINDINGS: A significant group effect was found (P<0.05) for all four co-activation indices. Principal pattern one captured the amplitude and general shape of activity throughout the entire stance phase. ANOVA revealed a significant (P<0.05) group by muscle interaction for the principal pattern scores. Significant differences were found among all three groups and between the two osteoarthritic groups for both measures. INTERPRETATION: The co activation indices and principal patterns identified that lateral site differences occurred among all three groups with medial site differences between the two osteoarthritic groups. These findings suggest that measures of muscle co activity provide additional information related to knee osteoarthritis severity. PMID- 19303180 TI - The efficacy of a removable vacuum-cushioned cast replacement system in reducing plantar forefoot pressures in diabetic patients. AB - BACKGROUND: The purpose of this study was to determine the plantar forefoot offloading efficacy of a new prefabricated vacuum-cushioned cast replacement system designed for foot ulcer treatment in neuropathic diabetic patients. METHODS: Fifteen diabetic subjects with peripheral neuropathy underwent in-shoe plantar pressure assessment while walking in five different footwear types: a standard vacuum-cushioned system with instantly moldable vacuum cushion and roller outsole, two modified vacuum-cushioned systems, one with flat surface vacuum cushion and one with flat outsole, a forefoot offloading shoe, and a control shoe. Regional peak pressures, pressure-time integrals, and inter regional load transfers were calculated to determine the mechanical action of the footwear. Perceived walking comfort was tested using a 10-point visual analogue scale. FINDINGS: Forefoot peak pressures and pressure-time integrals were significantly lower (by 41-56%) in the vacuum-cushioned system compared to control. Compared to the forefoot offloading shoe, the vacuum-cushioned system showed significantly higher metatarsal head peak pressures, similar metatarsal head pressure-time integrals, and significantly lower hallux peak pressures and pressure-time integrals. A major transfer of forefoot load to the midfoot explained the offloading efficacy of the vacuum-cushioned system. Few significant differences were present between the modified and standard vacuum-cushioned systems. Perceived walking comfort was significantly higher in the vacuum cushioned system (score 6.6) than in the forefoot offloading shoe (score 3.4). INTERPRETATION: The results showed that the vacuum-cushioned cast replacement system was effective in offloading the plantar forefoot of diabetic patients at risk for ulceration, although the contribution of the instantly moldable vacuum cushion and roller outsole were relatively small. The combined peak pressure, pressure-time integral and walking comfort results indicate that the vacuum cushioned system may be a useful alternative for the forefoot offloading shoe in offloading the plantar diabetic forefoot. PMID- 19303181 TI - Mechanical comparison of biodegradable femoral fixation devices for hamstring tendon graft--a biomechanical study in a porcine model. AB - BACKGROUND: Initial fixation strength is critical for the early post-operative rehabilitation of patients with anterior cruciate ligament reconstructions. However, even the best femoral fixation devices remain controversial. We compared the biomechanical characteristics of tendon grafts fixed by different biodegradable femoral fixation devices following anterior cruciate ligament reconstruction. METHODS: The Bio-TransFix, Rigidfix, Bioscrew with EndoPearl augmentation and Bioscrew devices were used to fix porcine flexor digitorum profundus tendon grafts in 32 porcine femora. Displacement of each tendon graft was evaluated after cyclic loading testing. Stiffness, ultimate failure load and failure mode of these fixation devices were measured with load-to-failure testing. FINDINGS: The displacement of the femur-graft-cement complex in response to cyclic loading was lower (P<0.05) for the Bio-TransFix than the Rigidfix, Bioscrew with EndoPearl augmentation, and Bioscrew groups. The fixation stiffness values of the Rigidfix and the Bioscrew were significantly greater (P<0.05) than that of the Bio-TransFix. The ultimate failure load was significantly greater for the Bio-TransFix and the Rigidfix than the Bioscrew with EndoPearl augmentation or the Bioscrew (P<0.05). INTERPRETATION: The Bio-TransFix provided the least graft displacement under cyclic loading. However, this device gave less stability. The Rigidfix device provided better stability and stiffness of the tendon graft among those fixation devices that showed no significant differences in graft displacement under cyclic loading. However, no single fixation device provided less displacement along with a larger failure load and stiffness in this study. PMID- 19303182 TI - Negative responses of Collembola in a forest soil (Alptal, Switzerland) under experimentally increased N deposition. AB - The response of specific groups of organisms, like Collembola to atmospheric nitrogen (N) deposition is still scarcely known. We investigated the Collembola community in a subalpine forest (Alptal, Switzerland) as subjected for 12 years to an experimentally increased N deposition (+25 on top of ambient 12 kg N ha(-1) year(-1)). In the 0-5 cm soil layer, there was a tendency of total Collembola densities to be lower in N-treated than in control plots. The density of Isotomiella minor, the most abundant species, was significantly reduced by the N addition. A tendency of lower Collembola group richness was observed in N-treated plots. The Density-Group index (dDG) showed a significant reduction of community diversity, but the Shannon-Wiener index (H') was not significantly affected by the N addition. The Collembola community can be considered as a bioindicator of N inputs exceeding the biological needs, namely, soil N saturation. PMID- 19303183 TI - Posttraumatic stress disorder and Operations Enduring Freedom and Iraqi Freedom: progress in a time of controversy. PMID- 19303184 TI - Ethnicity and nativity status as determinants of perceived social support: testing the concept of familism. AB - Research has demonstrated a protective effect of social support on health. Social support is most often treated as an independent variable. However, as with disease risk factors, which are not randomly distributed, health-promoting resources such as social support are also systematically patterned. For example, in the USA, family support is thought to be high among Latinos, Mexican Americans in particular. Using data from the Project on Human Development in Chicago Neighborhoods, we explored the relationships between ethnicity/nativity status, socioeconomic status (SES) and perceived social support from family and friends. We also assessed the role of retention of culture-measured as primary language spoken at home-on social support. Finally, we tested whether SES moderated the relationship between ethnicity/nativity status and social support. Foreign and US born Latinos, most notably, foreign-born Mexicans, reported higher family support compared to non-Latino whites. Primary language spoken at home seems to account for the relationship between ethnicity/nativity and familial social support. Mexican-born and US-born Latino immigrants reported lower social support from family at higher levels of SES. Each ethnic minority group reported lower perception of friend support compared to non-Latino whites. There was a strong SES gradient in subjective support from friends with higher support reported among those with higher SES. This study provides evidence for the notion that Latinos in the USA, specifically foreign-born Mexicans, may rely on family ties for support more than do non-Latino whites. Findings also help identify ethnicity/nativity status, primary language spoken and SES as determinants of social support. Specifically, the higher familial social support found among Latino immigrants may be due to retention of culture. Effect modification by SES suggests that Latinos of lower and higher SES may differ with regard to the traditionally-held value of familism. PMID- 19303185 TI - Between- and within-ear congruency and laterality effects in an auditory semantic/emotional prosody conflict task. AB - The present study investigated the influence of within- and between-ear congruency on interference and laterality effects in an auditory semantic/prosodic conflict task. Participants were presented dichotically with words (e.g., mad, sad, glad) pronounced in either congruent or incongruent emotional tones (e.g., angry, happy, or sad) and identified a target word or emotion under one of two conditions. In the within-ear condition, the congruent or incongruent dimensions were bound within a single stimulus and therefore, presented to the same ear. In the between-ear condition, the two dimensions were split between two stimuli and, therefore, presented in separate ears. Findings indicated interference in both conditions. However, the expected right ear advantage (EA) for words and left EA for emotions were obtained only in the between-ear condition. Factors involved in producing interference and laterality effects in dichotic listening tasks are discussed. PMID- 19303186 TI - Polyphenols, oral health and disease: A review. AB - OBJECTIVE: Polyphenols (PPs) are reactive metabolites abundant in plant-derived foods, particularly fruits, seeds and leaves. In the tissues of the digestive tract, particularly the oral mucosa, active PPs reach the highest concentration. Although excessively high and monotonic intake may be toxic (such an effect is countered by salivary proline-rich proteins), PPs exert preventive activity against infectious and degenerative diseases and may also help prevent oral diseases, via mechanisms like antioxidant activity and neutralisation/modulation of human/bacterial/viral proteins/enzymes. This review seeks to investigate the preventive PP activity against oral diseases. METHODS: Studies published during the last 15 years were searched using MEDLINE, PUBMED, EMBASE. The quality of evidence was rated using the three-point scale used by the US Preventive Services Task Force to assess the merits of preventive measures. RESULTS: Consistent, well designed, epidemiological, animal and in vitro studies directly and indirectly support the preventive PP effect against oral cancer with good evidence. Consistent studies showing that PPs inactivate periodontal pathogens and increase antioxidant capacity of oral fluids, suggest a preventive effect against periodontal disease with fair evidence. Animal and in vitro studies showing that PPs decrease mutans streptococci virulence and dental plaque accumulation, suggest a preventive effect against dental caries with fair evidence. Meta analysing clinical trials on subjects refraining from oral hygiene, PP-containing mouthrinses decrease mutans streptococci level by almost 50% and Plaque Index score. CONCLUSIONS: Regular and frequent PP intake may help protect against oral cancer, while human studies are needed to confirm the promising preventive activity against periodontal disease and caries. PMID- 19303187 TI - Comment on "The application of ozone in dentistry: A systematic review of the literature". PMID- 19303188 TI - Intraosseous infusion systems in the prehospital setting. PMID- 19303189 TI - A case of the heart ruling the head: acute myocardial infarction presenting with thunderclap headache. PMID- 19303190 TI - Extraction, quantification and characterization of uterine magnetomyographic activity--a proof of concept case study. AB - OBJECTIVE: The objective was to extract, quantify and characterize the uterine magnetomyographic (MMG) signals that correspond to the electrophysiological activity of the uterus. METHODS: Transabdominal MMG recordings with high spatial temporal resolution were performed with the use of the 151 non-invasive magnetic sensor system. The extraction, quantification and characterization procedures were developed and applied to representative MMG signals that were recorded from a pregnant woman at regular intervals starting at 37 weeks of gestation until the subject reached active labor. RESULTS: Multiple MMG recordings were successfully performed on the subject before she went into active labor. The extracted MMG burst activity showed a statistically significant correlation (r=0.2; p<0.001) with the contractile events perceived by mothers. The time-frequency analysis of the burst activity showed a power shift towards higher-frequency at 48 h before the subject went into active labor as compared to earlier recordings. Further there was a gradual increase in the synchrony in the higher-frequency band as the subject reached close to active labor. CONCLUSIONS: The non-invasive recording of the magnetic signals of pregnant uterus with high spatial-temporal resolution can provide an insight into the preparatory phase of labor and has the potential of predicting term and preterm labor. PMID- 19303191 TI - Biomechanics of the fetal membrane prior to mechanical failure: review and implications. AB - Annually, premature birth is a major public health problem accounting for over 13,000 deaths and 30,000 surviving infants with life-long morbidity. Preterm premature rupture of the membranes is the initiating event leading to preterm birth of 40% of these premature infants. Fetal membrane (FM) rupture is a catastrophic tissue failure, a unique event in normal physiology; other tissue failures (bone breaks, aneurism ruptures) are pathological processes. The mechanisms which cause FM failure and thereby rupture are not understood. A full understanding of FM failure process requires a complete characterization of structural and biomechanical behavior at near/full term under sub-failure (forces well below that which induce rupture) and failure conditions as well as elucidating the biological factors which modulate its failure. The relatively, highly loaded state of the FM in vivo may also facilitate its susceptibility to enzymatic degradation, which was shown to be augmented with increased load in collagenous tissues. Indeed, this last observation may help to provide the link between biomechanical degradation and premature mechanical failure in the FM. This integrated approach will further the understanding of this unique physiological event and thereby provide insight into how to anticipate and when appropriate, intervene to prevent preterm FM rupture. PMID- 19303192 TI - A short review of twin pregnancy and how oxytocin receptor expression may differ in multiple pregnancy. AB - During a multiple pregnancy, the mother and her fetuses are exposed to a variety of risks during both pregnancy and labour. The most notable of these risks is that of pre-term labour and its associated sequelae. Whilst much research has been directed towards understanding the mechanisms of uterine contractility, very little research has focussed on how contractility in multiple pregnancy differs from contractility in the singleton pregnancy. The aim of this paper is to review the changing prevalence and risks of a twin pregnancy, as well as reviewing what is known about myometrium from multiple pregnancies. The paper ends by discussing how oxytocin receptor expression may differ in twin pregnancy, based on the evidence of animal models, as well as presenting our own evidence of how oxytocin affects myometrium from twin pregnancies. We highlight the lack of the basic information needed to characterize human myometrium in twin pregnancies. Of particular note is the lack of supporting data for the hypothesis that stretch is responsible for earlier activation of the uterus in multiple pregnancy. New hypotheses based on increased experimental work are called for. Such information may throw light on specific mechanisms leading to the increased incidence of pre term delivery in twins. PMID- 19303193 TI - The use of an atmospheric dispersion model to determine influence regions in the Prince George, B.C. airshed from the burning of open wood waste piles. AB - A means of determining air emission source regions adversely influencing the city of Prince George, British Columbia, Canada from potential burning of isolated piles of mountain pine beetle-killed lodge pole pine is presented. The analysis uses the CALPUFF atmospheric dispersion model to identify safe burning regions based on atmospheric stability and wind direction. Model results show that the location and extent of influence regions is sensitive to wind speed, wind direction, atmospheric stability and a threshold used to quantify excessive concentrations. A concentration threshold based on the Canada Wide PM(2.5) Standard is used to delineate the influence regions while Environment Canada's (EC) daily ventilation index (VI) is used to quantify local atmospheric stability. Results from the analysis, to be used by air quality meteorologists in assessing daily requests for burning permits, are presented as a series of maps delineating acceptable burning locations for sources placed at various distances from the city center and under different ventilation conditions. The results show that no burning should be allowed within 10 km of the city center; under poor ventilation conditions, no burning should be allowed within 20 km of the city center; under good ventilation conditions, burning can be allowed within 10-15 km of the city center; under good to fair ventilation conditions, burning can be allowed beyond 15 km of the city center; and if the wind direction can be reliably forecast, burning can be allowed between 5 and 10 km downwind of the city center under good ventilation conditions. PMID- 19303194 TI - Public support for river restoration. A mixed-method study into local residents' support for and framing of river management and ecological restoration in the Dutch floodplains. AB - In many European countries, accommodating water has become the dominant paradigm in river management. In the Netherlands, extensive river restoration projects are being implemented, many of which draw serious opposition from the public. To investigate the causes of such opposition, a comprehensive study of public attitudes towards river restoration was conducted in three floodplains, both before and after river restoration. The study combined quantitative questionnaires (N=562) with open interviews (N=29). This paper describes how local residents perceive the effects of river restoration on landscape quality and how residents and protest groups use landscape quality in combination with other arguments to strategically frame river management policies. Results show that measurement of the perceived outcomes of nature restoration needs to be complemented by a more dynamic type of research, focusing on the social processes of the framing of restoration plans. Theoretically, the paper aims to contribute to the development of a rigorous research strategy to study framing processes in environmental management, using a mixed-methods approach. In general, local residents are supportive of river restoration projects. Although restoration may diminish feelings of attachment to an area, for most people this negative effect is compensated by the positive effects on scenic beauty and perceived protection from flooding. However, these positive effects may become contested because of the active framing of river restoration by protest groups. Residents use three distinct frames to give meaning to river restoration projects: (i) an attachment frame, focusing on cultural heritage and place attachment (ii) an attractive nature frame, focusing on nature as attractive living space and the intrinsic value of nature (iii) a rurality frame, focusing on rural values, agriculture and cultural heritage. Resistance to river restoration plans stems from the attachment and rurality frames. People using these frames challenge safety arguments for river restoration and highlight potential threats to sense of place and to agriculture. In the areas surveyed, the project initiator's focus on biodiversity and safety did not resonate very well among the local community, because of their diverging views on nature. Practical implications of the study include the need to incorporate public perception into river restoration projects and the potential for project initiators to form strategic alliances with local residents to promote ecological restoration in combination with river restoration. PMID- 19303195 TI - Bioleaching of heavy metals from sewage sludge: a review. AB - During the treatment of sewage, a huge volume of sludge is generated, which is disposed of on land as soil fertilizer/conditioner due to the presence of nitrogen, phosphorus, potassium and other nutrients. However, the presence of toxic heavy metals and other toxic compounds in the sludge restricts its use as a fertilizer. Over the years, bioleaching has been developed as an environmentally friendly and cost-effective technology for the removal of heavy metals from the sludge. The present paper gives an overview of the various bioleaching studies carried out in different modes of operation. The various important aspects such as pathogen destruction, odor reduction and metal recovery from acidic leachate also have been discussed. Further, a detailed discussion was made on the various technical problems associated with the bioleaching process, which need to be addressed while developing the process on a larger scale. PMID- 19303196 TI - Editorial comment on: Prognostic parameters, complications, and oncologic and functional outcome of salvage radical prostatectomy for locally recurrent prostate cancer after 21st-century radiotherapy. PMID- 19303197 TI - Prognostic parameters, complications, and oncologic and functional outcome of salvage radical prostatectomy for locally recurrent prostate cancer after 21st century radiotherapy. AB - BACKGROUND: Salvage radical prostatectomy (SRP) for radiorecurrent prostate cancer (PCa) is a second local treatment with curative intent in patients with true organ-confined recurrent PCa. OBJECTIVE: We evaluated preoperative prognostic risk factors to predict organ-confined, locally recurrent PCa after primary radiotherapy (RT). DESIGN, SETTING, AND PARTICIPANTS: Fifty-five men with biopsy-proven, locally recurrent PCa underwent SRP and extended pelvic lymph node dissection (ePLND) after external-beam radiotherapy (EBRT) or low- or high-dose brachytherapy. MEASUREMENTS: Prostate-specific antigen (PSA), clinical stage, biopsy Gleason score prior to RT and SRP, PSA nadir, time to recurrence, PSA doubling time (PSA DT), PSA prior to surgery, and pathohistology of the SRP specimen were analysed to predict organ-confined recurrent disease. Uni- and multivariate statistical analysis was performed. RESULTS AND LIMITATIONS: Forty (72.7%) and 15 (27.3%) patients demonstrated organ-confined and locally advanced PCa, respectively. Eleven patients (20%) and seven patients (12.7%) had lymph node metastases and positive surgical margins (PSM), respectively. On multivariate analysis, biopsy Gleason score prior to SRP (p=0.02), <50% positive biopsy cores (p=0.001), PSA DT >12 mo (p=0.001), and low-dose brachytherapy (p=0.001) were significant predictors of organ-confined PCa with negative surgical margins (NSM). Limitations of the study are its retrospective nature and the relatively low number of patients. CONCLUSIONS: SRP is a surgically challenging but effective secondary local treatment of radiorecurrent PCa with curative intent. The identified predictive parameters will help to select patients most suitable for SRP with long-term cure and good functional outcome. PMID- 19303198 TI - Editorial comment on: Prognostic parameters, complications, and oncologic and functional outcome of salvage radical prostatectomy for locally recurrent prostate cancer after 21st-century radiotherapy. PMID- 19303199 TI - Editorial comment on: erectile implants in female-to-male transsexuals: our experience in 129 patients. PMID- 19303200 TI - Erectile implants in female-to-male transsexuals: our experience in 129 patients. AB - BACKGROUND: The combination of a neourethra and erection prosthesis in a single neophallus in the female-to-male transsexual remains a challenge. No good data are available on this subject. OBJECTIVE: To report the outcome in 129 female-to male transsexuals with a neophallus after the implantation of an erectile prosthesis. DESIGN, SETTING, AND PARTICIPANTS: From March 1996 until October 2007, 129 female-to-male transsexuals with a neophallus underwent the implantation of an erectile prosthesis. The mean follow-up was 30.2 mo (range: 0 132 mo). INTERVENTION: A Dynaflex prosthesis was implanted initially in 9 patients, a three-piece hydraulic device (AMS CX or AMS CXM) in 50 patients, and a CX Inhibizone, Ambicor, and Coloplast/Mentor prosthesis in 17, 47, and 6 patients, respectively. MEASUREMENTS: Data on outcome in these patients were retrospectively evaluated. RESULTS AND LIMITATIONS: Of 129 patients, 76 patients (58.9%) still have their original implant in place. Fifty-three patients (41.1%) needed to undergo either removal or revision of the prosthesis due to infection, erosion, dysfunction, or leak. Forty-one patients underwent a replacement of the prosthesis, nine needed a second revision, five needed a third revision, and one patient needed a fourth revision of prosthesis. Malposition of prosthesis was corrected by surgical repositioning so that removal or revision could be avoided. Of 185 prostheses used in 129 patients, 108 (58.4%) still remain in place, with a total infection rate of 11.9%, a total protrusion rate of 8.1%, a total prosthesis leak rate of 9.2%, a total dysfunction rate of 13%, and a total malposition rate of 14.6%. The period of follow-up in the more recent types of prostheses (Ambicor, Coloplast/Mentor) is much shorter; therefore, comparison with earlier types is difficult to make. CONCLUSIONS: Despite high complication rates, implantation of a hydraulic erectile prosthesis remains the best option for achieving the possibility of sexual intercourse in female-to-male transsexuals. PMID- 19303201 TI - Intermittent androgen deprivation therapy: conventional wisdom versus evidence. PMID- 19303202 TI - Re: Ofer Yossepowitch, Anders Bjartell, James A. Eastham, et al. Positive surgical margins in radical prostatectomy: outlining the problem and its long term consequences. Eur Urol 2009;55:87-99. PMID- 19303203 TI - Why doesn't high-dose chemotherapy improve survival, as was expected, in advanced or relapsed germ cell tumours? PMID- 19303204 TI - Editorial comment on: erectile implants in female-to-male transsexuals: our experience in 129 patients. PMID- 19303205 TI - CSF levels of cytokines in neuro-Behcet's disease. AB - BACKGROUND: Neurological manifestations of Behcet's disease (neuro-Behcet's disease) present in 5-30% of patients. Although cytokines play a pivotal role in pathogenesis of Behcet's disease, published studies about the cerebrospinal fluid (CSF) levels of cytokines in neuro-Behcet's disease are scanty. METHODS: Nine patients with active parenchymal, one patient with non-parenchymal neuro-Behcet's disease, six patients with headache attributed to Behcet's disease, 13 patients with viral meningitis, and 19 healthy controls were recruited. Interleukin 6, 8, 10, tumor necrotic factor-alpha, and interferon-gamma were measured in the CSF using enzyme-linked immunosorbent assay method. RESULTS: Patients with viral meningitis had significantly higher levels of all investigated cytokines except for interferon-gamma in comparison with the patients with parenchymal neuro Behcet's disease, headache attributed to Behcet's disease and controls (P values <0.05). CSF interleukin 6 was significantly higher in patients with parenchymal neuro-Behcet's disease in comparison with the controls (P=0.025). CSF levels of investigated cytokines had no significant difference between patients with headache attributed to Behcet's disease and controls (P values >0.05). Patients with headache attributed to BD and patients with parenchymal NBD had no significant difference in measured cytokines (P values >0.05). CONCLUSION: In contrast to some previous studies, our investigation showed loss of analogy between CSF cytokine profiles of patients with parenchymal neuro-Behcet's disease and viral meningitis. Also we postulated a crucial role for interleukin 6 in immunopathogenesis of neuro-Behcet's disease. PMID- 19303206 TI - Cyclin D1 negatively regulates the expression of differentiation genes in HT-29 M6 mucus-secreting colon cancer cells. AB - HT-29 M6 colon cancer cells differentiate to a mucus-secreting phenotype in culture. We found that the pattern of cyclin D1 expression in HT-29 M6 cells did not correlate with instances of cell proliferation but was specifically induced during a dedifferentiation process following disaggregation of epithelial cell layers, even under conditions that did not allow cell cycle reentrance. Interestingly, ectopic expression of cyclin D1 in differentiated cells led to the inhibition of the transcriptional activity of differentiation gene promoters, such as the mucin MUC1. We thus propose that the overexpression of cyclin D1 found in colon cancer favours tumour dedifferentiation as one mechanism of tumour progression. PMID- 19303207 TI - Cell biology of the movement of breast cancer cells: intracellular signalling and the actin cytoskeleton. AB - Cell motility is a critical step in cancer invasion and metastasis that must be unravelled to gain an appropriate understanding of the behaviour of cancer cells. A broad spectrum of motility mechanisms that facilitate invasion of extramammary tissues and metastasis exists in breast cancer cells (e.g. reorganization of the actin cytoskeleton, regulation of focal adhesion, changes in response to a different microenvironment, epithelial mesenchymal transition, and control of membrane proteins through endocytosis). These cellular responses are tightly regulated by intracellular signalling pathways evoked by humoral factors that include growth factors, chemokines, and cytokines. Learning more about the cellular and molecular basis of these different motility programmes will aid in the development of treatments for breast cancer invasion and metastasis. This review of recent literature focuses on aspects of cell biology related to motility and metastasis, and suggests some directions for future breast cancer research. PMID- 19303208 TI - HDAC inhibitors: magic bullets, dirty drugs or just another targeted therapy. PMID- 19303209 TI - Synthesis of nesquehonite by reaction of gaseous CO2 with Mg chloride solution: its potential role in the sequestration of carbon dioxide. AB - In this paper is reported a novel method to synthesize nesquehonite, MgCO(3) x 3H(2)O, via reaction of a flux of CO(2) with Mg chloride solution at 20+/-2 degrees C. The reaction rate is rapid, with carbonate deposition almost complete in about 10 min. The full characterization of the product of synthesis has been performed to investigate its potential role as a "CO(2)-sequestering medium" and a means of disposing Mg-rich wastewater. We investigated the nesquehonite synthesized using SEM, XRD, FTIR and thermal analysis. The thermodynamic and chemical stability of this low-temperature hydrated carbonate of Mg and its possible transformation products make our method a promising complementary solution to other methods of CO(2) sequestration. Carbonation via magnesium chloride aqueous solutions at standard conditions represents a simple and permanent method of trapping CO(2). It could be applied at point sources of CO(2) emission and could involve rejected brine from desalination plants and other saline aqueous wastes (i.e., "produced water"). The likelihood of using the resulting nesquehonite and the by-products of the process in a large number of applications makes our method an even more attractive solution. PMID- 19303210 TI - Removal of azo and anthraquinone reactive dyes from industrial wastewaters using MgO nanoparticles. AB - In the present investigation, a porous MgO powder was synthesized and tested for the removal of dyes from aqueous solution. The size of the MgO particles was in the range of 38-44 nm, with an average specific surface area of 153.7 m(2)/g. Adsorption of reactive blue 19 and reactive red 198 was conducted to model azo and anthraquinone dyes at various MgO dosages, dye concentrations, solution pHs and contact times in a batch reactor. Experimental results indicate that the prepared MgO powder can remove more than 98% of both dyes under optimum operational conditions of a dosage of 0.2g, pH 8 and a contact time of 5 min for initial dye concentrations of 50-300 mg/L. The isotherm evaluations revealed that the Langmuir model attained better fits to the experimental equilibrium data than the Freundlich model. The maximum predicted adsorption capacities were 166.7 and 123.5mg of dye per gram of adsorbent for RB 19 and RR 198, respectively. In addition, adsorption kinetic data followed a pseudo-second-order rate for both tested dyes. PMID- 19303211 TI - Simultaneous removal of coexistent heavy metals from simulated urban stormwater using four sorbents: a porous iron sorbent and its mixtures with zeolite and crystal gravel. AB - The selectivity sequence and removal of coexistent heavy metals (namely As, Cd, Cr, Cu, Ni and Zn) in synthetic urban stormwater runoff were investigated by adsorption onto a porous iron sorbent (namely P1) and its mixtures with zeolite and crystal gravel, respectively (namely P2, P3, and P4). A batch method was employed to simulate the sorption processes. The geochemical model PHREEQC was used to calculate the metals' species and saturation data for elucidating the sorption data. The equilibrium data demonstrated a good fit with the Freundlich model and showed affinity in the orders: Cd>Zn>Ni>Cu>As>Cr (sorbents P1, P3 and P4) and Cd>Zn>Ni>As>Cu>Cr (sorbent P2). In addition to this, Calculated Distribution Coefficient (K(d)) values were used to compare the overall heavy metal removal efficiencies of the sorbents, which, in decreasing order, was found to be P4>P1>P2>P3. In comparing these four commercial sorbents, sorbent P4 represents a promising material for treatment of urban stormwater runoff containing mixed heavy metals. PMID- 19303212 TI - Aerobic degradation of nitrobenzene by immobilization of Rhodotorula mucilaginosa in polyurethane foam. AB - Rhodotorula mucilaginosa Z1 capable of degrading nitrobenzene was immobilized in polyurethane foam. The nitrobenzene-degrading capacity of immobilized cells was compared to free cells in batches in shaken culture. Effects of pH and temperature on the nitrobenzene degradation showed that polyurethane-immobilized Z1 had higher tolerances toward acid, alkali, and heat than those of free cells. Kinetic studies revealed that higher concentrations of nitrobenzene were better tolerated and more quickly degraded by polyurethane-immobilized Z1 than by free cells. Moreover, the ability of polyurethane-immobilized Z1 to resist nitrobenzene shock load was enhanced. Experiments on the nitrobenzene degradation in different concentrations of NaCl and in the presence of phenol or aniline demonstrated that polyurethane-immobilized Z1 exhibited higher tolerance toward salinity and toxic chemicals than those of free cells. Immobilization therefore could be a promising method for treating nitrobenzene industrial wastewater. This is the first report on the degradation of nitrobenzene by a polyurethane immobilized yeast strain. PMID- 19303213 TI - Gastrointestinal parasites in dogs from the Island of St. Pierre off the south coast of Newfoundland. AB - The present work was performed to survey the gastrointestinal parasites of domestic dogs (Canis lupus familiaris) and also to determine if any were infected with Angiostrongylus vasorum (French Heartworm), on the French Island of St. Pierre off the south coast of Newfoundland. A total of 57 fecal samples were collected and examined for intestinal parasites. The overall prevalence of parasitism was 57.9% and the six species found were: Uncinaria stenocephala/Ancylostoma caninum (47.4%), Toxocara canis (22.8%), Isospora canis (8.8%), Trichuris vulpis (7.0%), and Alaria canis (1.8%). There was no significant difference in overall prevalence between genders, except for T. canis, which was more common in female dogs than male dogs (p<0.05). PMID- 19303214 TI - Genetic variability among Schistosoma japonicum isolates from different endemic regions in China revealed by sequences of three mitochondrial DNA genes. AB - The present study examined sequence variation in three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 3 (cox3), NADH dehydrogenase subunits 4 and 5 (nad4 and nad5), among Schistosoma japonicum isolates from different endemic regions in China, and their phylogenetic relationships were re constructed. A portion of the cox3 gene (pcox3), a portion of the nad4 and nad5 genes (pnad4 and pnad5) were amplified separately from individual trematodes by polymerase chain reaction (PCR) and the amplicons were subjected to direct sequencing. In the mountainous areas, sequence variations between parasites from Yunnan and those from Sichuan were 0.3% for pcox3, 0.0-0.1% for pnad4, and 0.0 0.2% for pnad5. In the lake/marshland areas, sequence variations between male and female parasites among different geographical locations were 0.0-0.3% for pcox3, 0.0-0.7% for pnad4, and 0.0-1.6% for pnad5. Sequence variations between S. japonicum from mountainous areas and those from lake/marshland areas were 0.0 0.5% for pcox3, 0.0-0.7% for pnad4, and 0.0-1.6% for pnad5. Phylogenetic analyses based on the combined sequences of pcox3, pnad4 and pnad5 revealed that S. japonicum isolates from mountainous areas (Yunnan and Sichuan provinces) clustered together. For isolates from the lake/marshland areas, isolates from Anhui and Jiangsu provinces clustered together and was sister to samples from Jiangxi province, while isolates from Hubei and Zhejiang province clustered together. However, isolates from different geographical locations in Hunan province were in different clades. These findings demonstrated the usefulness and attributes of the three mtDNA sequences for population genetic studies of S. japonicum, and have implications for studying population biology, molecular epidemiology, and genetic structure of S. japonicum, as well as for the effective control of schistosomiasis. PMID- 19303215 TI - Crab-eating fox (Cerdocyon thous), a South American canid, as a definitive host for Hammondia heydorni. AB - Hammondia heydorni is a cyst forming coccidia closely related to other apicomplexans, such as Toxoplasma gondii, Neospora caninum and Hammondia hammondi with a two-host life cycle. Dogs and other canids as red foxes (Vulpes vulpes) and coyotes (Canis latrans) may serve as definitive hosts for H. heydorni. Sporulated oocysts are infective for cattle, sheep and goats, which may serve as intermediate hosts. Herein, we describe the ability of crab-eating fox (Cerdocyon thous), a wild carnivore that is commonly found from northern Argentina to northern South America, to serve as definitive host of H. heydorni. The whole masseter muscle and brain from two 2-year-old bovines were collected, minced and pooled together for the fox infection. The bovine pooled tissues were equally administered to four foxes, in two consecutive days. Two foxes shed subspherical unsporulated oocysts measuring 10-15microm, after 8 and 9 days post-infection, respectively. One of the foxes eliminated oocysts for 5 days, while the other fox shed oocysts for 9 days. A DNA sample of oocysts detected at each day of oocyst elimination was tested by two PCRs, one of them carried out employing primers directed to the common toxoplasmatiid 18S and 5.8S ribosomal RNA coding genes (PCR-ITS1) and the other based on heat-shock protein 70kDa coding gene (PCR HSP70). These samples were also submitted to a N. caninum specific nested-PCR protocol based on a N. caninum specific gene (Nc5-nPCR). All of them were positive by PCR-ITS1 and PCR-HSP70 but negative by Nc5-nPCR. The PCR-ITS1 and PCR HSP70 nucleotide sequences amplified from the oocysts shed by the foxes revealed 100% identity with homologous sequences of H. heydorni. In conclusion, it is clear that H. heydorni also uses the crab-eating fox as a definitive host. The crab-eating fox is usually reported to live in close contact with livestock in several regions of Brazil. Therefore, it is reasonable to infer that such carnivores may play an important role in the sylvatic and domestic cycles of H. heydorni infection. PMID- 19303216 TI - Paralytic ileus and subsequent death caused by enteric parasite, Strongyloides papillosus, in Mongolian gerbils. AB - Adult worms of Strongyloides papillosus were surgically implanted into the duodenum and successfully established in Mongolian gerbils (Meriones unguiculatus). These worms persisted in the small intestine for at least four days after implantation. Following decreased fecal output, however, increased death rate and decreased survival time were observed. The increase in death rate and the decrease in survival time correlated with the increase in the number of implanted adult worms. Animals were then intraperitoneally inoculated with extracts from adult S. papillosus, and the pathogenetic effects of gastrointestinal (GI) motility were assessed by contrast radiography after oral administration of barium sulfate. Paralytic ileus was observed in the GI tracts of Mongolian gerbils and these symptoms intensified with increasing inoculation of adult worm extract. The results suggest that paralytic ileus underlies the subsequent death observed in Mongolian gerbils after implantation of adult S. papillosus. Furthermore, experimental infection with S. papillosus in Mongolian gerbils will provide a good model for laboratory investigations into GI motor disturbances in animals and humans caused by parasites. PMID- 19303217 TI - Interleukin-8 is elevated in cerebrospinal fluid following high-voltage electrical injury with late-onset paraplegia suggesting neuronal damage at the microlevel as causative factor. PMID- 19303218 TI - Pentoxifylline modifies three-dimensional collagen lattice model contraction and expression of collagen types I and III by human fibroblasts derived from post burn hypertrophic scars and from normal skin. AB - Fibroblasts are thought to be partially responsible for the persisting contractile forces that result in burn contractures. Using a monolayer cell culture and fibroblast populated collagen lattice (FPCL) three-dimensional model we subjected hypertrophic scar and non-cicatricial fibroblasts to the antifibrogenic agent pentoxifylline (PTF - 1mg/mL) in order to reduce proliferation, collagen types I and III synthesis and model contraction. Fibroblasts were isolated from post-burn hypertrophic scars (HSHF) and non scarred skin (NHF). Cells were grown in monolayers or incorporated into FPCL's and exposed to PTF. In monolayer, cell number proliferation was reduced (46.35% in HSHF group and 37.73% in NHF group, p<0.0001). PTF selectively inhibited collagen III synthesis in the HSHF group while inhibition was more evident to type I collagen synthesis in the NHF group. PTF also reduced contraction in both (HSHF and NHF) FPCL. PMID- 19303219 TI - Client and service characteristics associated with addiction treatment completion of clients with co-occurring disorders. AB - The study examines client and service characteristics of addiction treatment completers and non-completers with co-occurring disorders (COD). On demographic variables, completers were more likely to be male and homeless. In the psychiatric domain, a greater proportion of completers received diagnoses of depression and generalized anxiety disorder, whereas non-completers were more often diagnosed with bipolar disorder and posttraumatic stress disorder. No group differences were found in client-reported psychiatric symptom severity; however, non-completers were rated by clinicians as having more severe symptoms in the areas of interpersonal sensitivity, depression, and hostility. In the area of substance use patterns, no differences were found in primary substance of abuse but completers reported more days of use during the month prior to treatment. Completers also had a greater history of both prior detox and non-detox treatment. At discharge, completers achieved higher rates of past month abstinence and AA attendance, but no differences were found in length of stay in treatment. Examination of recovery support services utilization revealed that completers more often received peer mentoring services. Greater proportions of the non-completer group received educational support, clothing, medical care, and employment assistance. These results suggest that future studies are needed in examining possible differential treatment response by diagnostic category and the potential role of peer mentoring in enhancing addiction treatment completion of COD clients. PMID- 19303220 TI - An association between geomagnetic activity and dream bizarreness. AB - Daily disturbances of the earth's magnetic field produce variations in geomagnetic activity (GMA) that are reportedly associated with widespread effects on human health and behaviour. Some of these effects could be mediated by an established influence of GMA on the secretion of melatonin. There is evidence from unrelated research that melatonin influences dream bizarreness, and it is hypothesised here that there is an association between GMA and dream bizarreness. Also reported is a preliminary test of this hypothesis, a case study in which the dreams recorded over 6.5 years by a young adult male were analysed. Reports of dreams from the second of two consecutive days of either low or high GMA (K index sum < or =6 or > or = 28) were self-rated for bizarreness on a 1-5 scale. Dreams from low GMA periods (n=69, median bizarreness=4) were found to be significantly more bizarre than dreams from high GMA periods (n=85, median bizarreness=3; p=0.006), supporting the hypothesised association between GMA and dream bizarreness. Studies with larger samples are needed to verify this association, and to determine the extent to which melatonin may be involved. Establishing that there is an association between GMA and dream bizarreness would have relevance for neurophysiological theories of dreaming, and for models of psychotic symptoms resembling bizarre dream events. PMID- 19303221 TI - Quantitative assessment of red meat or processed meat consumption and kidney cancer. AB - OBJECTIVE: To conduct a quantitative assessment of red meat or processed meat consumption and kidney cancer. METHODS: We extracted data from 12 case-control studies, three cohort studies, and the Pooling Project of Diet and Cancer publication for which 13 international cohorts were evaluated. Random effects meta-analysis models were used to calculate summary relative risk estimates (SRRE) based on high vs. low intake values. Sensitivity and influence analyses were conducted, including assessments of heterogeneity. RESULTS: The SRRE for all studies that reported results for red meat (included variables labeled 'red meat' or single red meat items, such as beef, pork, or liver) was 1.12 (95% CI: 0.98 1.29; p-value for heterogeneity=0.015), and the SRRE using only data from prospective cohorts was 1.02 (95% CI: 0.91-1.15) with minimal heterogeneity (p=0.741). Similarly, in a meta-analysis of the five studies that simultaneously adjusted for smoking, BMI, and total energy intake, the SRRE for red meat was 1.02 (95% CI: 0.91-1.15). No significant association was observed in the meta analysis of processed meat consumption (SRRE=1.07; 95% CI: 0.94-1.23), although a significant association was observed when only data from cohort studies were analyzed (SRRE=1.19; 95% CI: 1.03-1.37). CONCLUSIONS: Although many of the summary results were positive, all were weak in magnitude, most were not statistically significant, and associations were attenuated among studies that adjusted for key potential confounding factors. In summary, the findings of this meta-analysis are not supportive of an independent relation between red or processed meat intake and kidney cancer. PMID- 19303222 TI - [Abdominal migration of a levonorgestrel-releasing intrauterine device. Case report and review of the literature]. AB - We report the case of a 27-year-old woman in amenorrhea after the installation of a levonorgestrel releasing intra-uterine device, 3 years previously. In front of pelvic continuing pains, paraclinic explorations diagnosed the DIU-LNG in intra abdominal situation. An exploratory laparoscopy was realized and allowed its ablation. The authors discuss the systemic distribution of the levonorgestrel and the impact of the DIU-LNG in intra-abdominal situation by a review of the literature. PMID- 19303223 TI - [A rare case of placental chorioangioma associated with neonatal disseminated hemangiomatosis]. AB - Placental chorioangioma is a benign vascular tumor. Lesions larger than 4 cm may cause fetal and maternal complications. Its association with disseminated neonatal hemangiomatosis is rarely described. We report a case of a large chorioangioma associated with an hydrops foetalis and disseminated neonatal hemangiomatosis. The relationship between placental chorioangioma and hemangioma is briefly discussed. PMID- 19303224 TI - [Hygiene rules of gynecologic consultation]. AB - Nosocomial infections occur not only to hospitalized patients but also to outpatients treated in private practice. Infections are secondary to methicillin resistant staphylococcus aureus (MRSA), Prions, and Blood transmission agents. Simple hygiene measures such as hand wash, device sterilization, or disposible devices are the best preventive solutions. PMID- 19303225 TI - [Bilateral gestational macromastia: case report]. AB - PURPOSE: We report a case of bilateral gestationnal macromastia in order to discuss the pathogeny, the diagnosis and the treatment. METHODOLOGY: It was a case of a 33-year-old woman, admitted for a bilateral massive hypertrophy of the breast occurring on pregnancy and with progressive evolution. She had three pregnancies and one born-infant. Biological exams have shown a hyperprolactinemia. Pathological exam of the mammary biopsy had shown a benign hyperplasia. RESULTS: Medical treatment of our patient by bromocriptin was inefficient. She has had a bilateral mastectomy. She is waiting for mammary plastic surgery. CONCLUSION: Gravidic macromastia is a rare pathology whose etiology and treatment are much debated. Frequent recurrence after mammary reduction justify the mastectomy followed by prothesis. PMID- 19303226 TI - [Major alpha-thalassemia: antenatal diagnosis, case report and literature review]. AB - Homozygous alpha-thalassaemia or Bart's hydrops fetalis is a genetic disease with autosomal recessive transmission. The condition is lethal for the fetus because of hypoxia and anemia. For the mother there is an increased risk of the severe forms of preeclampsia and its complications. The diagnosis can be suspected in presence of suggestive ultrasonographic anomalies, where both parents come from South-East Asia or China. Confirmation is based on the identification of the typical deletions or mutation of the alpha globin gene by molecular genetics. We report a rare clinical case of Bart's hydrops fetalis diagnosed because of fetal growth retardation, fetal cardiomegaly and increased size of placenta on the 26 weeks fetal echography. This case underscores the need to include the alpha thalassemias in medical and midwifery education in countries where they were almost inexistent a generation ago. PMID- 19303227 TI - Comments to "Comparison of antibody responses after vaccination with two inactivated rabies vaccines" [Minke, J.M., et al., 2009. Vet. Microbiol. 133, 283 286]. PMID- 19303228 TI - Blood-positive illicit-drug findings: implications for cause-of-death certification, classification and coding. AB - National cause-of-death data are important for national health administration, international comparisons and epidemiological research. The process of compiling mortality statistics starts with determination of causes of death, continues with medical death certification and concludes in coding of causes and selection of the underlying cause of death at statistical office. This study assesses how unequivocally and specifically the toxicologically verified intoxication of cannabinoids, opiates, amphetamines and cocaine is represented in diagnostic entries on death certificates and the national cause-of-death database and, ultimately, in the cause-of-death statistics. Drug-positive deaths, i.e. deaths with blood-positive drug finding(s), the corresponding death certificates and the information entered in Statistics Finland's cause-of-death database were reviewed for the entire years of 2000, 2002 and 2004. Drug influence at the time of death may or may not be related to death. A drug-related condition was reported as the cause of death in 52% of cannabinoid-positive, in 81% of amphetamine-positive, in 98% of opiate-positive and in 100% of cocaine-positive deaths, calculated from the combined three-year material. At Statistics Finland, after validation of the reported information, the distribution was practically the same. From the cause of-death database, the specific drug-related diagnosis could be identified in 21% of cannabinoid-positive, in 89% of opiate-positive and in 57% of amphetamine positive deaths. The corresponding proportions of specific drug-related underlying causes in the cause-of-death statistics were even smaller for cannabinoids and amphetamines, 10% and 39%, respectively. In multiple-drug cases, identification was possible only if each drug had been assigned an additional drug-specific code from "T categories" of ICD-10 Chapter XIX. What is noteworthy, however, is that a third of cannabinoid-related and a quarter amphetamine-related cause-of-death diagnoses were assigned unspecific categories of ICD-10 in the multiple-cause database and, more notably, in cause-of-death statistics based on selected underlying causes. For the better specification of drug-related causes of death, we propose that the next ICD revision provide each drug with the code of its own, e.g. one comparable to ATC (Anatomical Therapeutic Chemical Classification Index) codes, to be used for its specification in all positions and combinations, or at least provide compatibility with the ATC's coding system. For classification and describing the trends of drug-related deaths, equal and specific definitions for drug-related deaths would also be needed. PMID- 19303229 TI - The neuropathology of infant subdural haemorrhage. AB - Subdural haemorrhage (SDH) in the infant has a different pattern from that seen in the older child and adult. It is usually a widespread, bilateral, thin film, unlike the thick, space-occupying and often unilateral clot seen in older children and adults after trauma. Whether both arise by the same mechanism is unknown, but it seems unlikely. Most SDH is said to be due to trauma but in infants there are other, atraumatic causes. Birth is also important; recent MRI studies show an incidence of almost 50% in asymptomatic neonates. Traumatic SDH is said to result from rupture of bridging veins but new insights into the anatomy of infant dura suggest a dural origin for thin film subdural bleeding in young babies. Acute SDH usually rapidly resolves, but sometimes develops into a chronic fluid collection. Healing of SDH is by formation of a granulating membrane which may confer vulnerability to rebleeding, either spontaneously or after an otherwise innocuous event. SDH has a particular significance as one of the features of the triad (together with retinal haemorrhage and encephalopathy) associated with non-accidental injury. As the possibility of non-accidental injury is often first raised by a radiologic report of subdural bleeding, it becomes critically important in the interpretation of the scan appearances to understand the unique physiology and anatomy of the infant dura. PMID- 19303230 TI - Sudden death associated with intravenous injection of toad extract. AB - A 24-year-old male died suddenly following the intravenous injection of what was believed to be the ring-derivate amphetamine 'ecstasy' (MDMA). Toxicological analyses of the victim's blood and the injected material, however, failed to reveal MDMA, but showed instead low levels of bufotenine, a tryptamine derivative alkaloid found in the secretions of various toads. In addition, resibufogenin, cinobufagin and bufalin, bufadienolides that are also found in toad venom, were identified in the injected material. While these substances also occur in certain South American plants, the finding of paracetamol, promethazine and diclofenac would be in keeping with ingredients found in the traditional Chinese herbal product Chan Su that derives from the skin glands and secretions of toads and that is often adulterated with standard pharmaceutical drugs. This case demonstrates the problems that users and sellers may encounter from the unknown composition of street drugs and herbal medicines, and the danger that may be incurred from the injection of such materials. It also shows the difficulties that may be associated with attempting to identify low levels of organic toxins in postmortem specimens necessitating a targeted screening approach guided by information obtained at the death scene. PMID- 19303231 TI - Prevalence of acute otitis media among children with pyrexia in a Nigerian hospital. AB - OBJECTIVE: This was to determine the prevalence of acute otitis media in children with pyrexia in an area with malaria endemicity and also the relevance of socio economic factors on AOM. METHODS: This prospective study was conducted between January 2004 and June 2005 at the emergency paediatric unit of the hospital among children aged 0-15 years presenting with pyrexia. All the children had full ENT examinations in addition to the evaluation by the emergency paediatricians. Diagnosis of AOM was based on history, examinations and otoscopic findings. RESULTS: Two-hundred children with pyrexia were seen and the age range was 3 months to 15 years (mean 4.73). The male/female ratio was 1.7:1.03 years; 32 had features of AOM. Two third of the children were from the low socio-economic class. CONCLUSION: AOM with late presentation in the suppurative stage of the disease is a common cause of pyrexia in children with a male preponderance and two third of the children were from the low socio-economic status. There is the need for otoscopic examinations of all pyretic children as the resultant hearing loss is related to difficulties in language acquisition in children below 2 years of age with effects on literacy and school achievement. PMID- 19303232 TI - [General practitioners' perception of the impact of corticosteroid-induced adverse events]. AB - BACKGROUND: General practitioners' perception of the discomfort their patients experience because of corticosteroid-induced adverse events is unknown. METHODS: An observational epidemiological study was conducted in September 2007. Eight hundred and sixty general practitioners belonging to the reseau Sentinelles were asked to complete an electronical questionnaire. The questionnaire aimed to assess their perception of discomfort induced by adverse events induced by a long term (i.e.,>or=3 months) corticosteroid therapy among their patients. Results were compared with the declaration made by 115 long-term corticosteroid treated patients followed in an internal medicine department. RESULTS: Two hundred and ninety-three general practitioners responded to the questionnaire (response rate: 34%). They were predominantly male (87%). Forty-eight percent of them reported 400 to 600 monthly visits. The mean length of corticosteroid therapy for patients was 44+/-38 months and the mean daily dosage was 15+/-14 mg. They suffered mainly from lupus erythematosus (33%) or giant cell arteritis (15%). The adverse events considered to be the most disturbing by patients were lipodystrophy (25%), followed by weight gain (18%) and neuropsychiatric complaints (16%). Physicians widely overestimated the discomfort caused by weight gain cited as the most disturbing adverse event by 59% of them and underestimated that induced by mood disorders cited as the most disturbing by only 3% of them. CONCLUSION: The discomfort caused by corticosteroid-induced neuropsychiatric adverse events are underestimated by general practitioners. PMID- 19303233 TI - Radiogenomics: creating a link between molecular diagnostics and diagnostic imaging. AB - Studies employing high-throughput biological techniques have recently contributed to an improved characterization of human cancers, allowing for novel sub classification, better diagnostic accuracy, and more precise prognostication. However, requirement of surgical procurement of tissue among other things limits the clinical application of such methods in everyday patient care. Radiographic imaging is routine in clinical practice but is currently histopathology based. The use of routine radiographic imaging provides a potential platform for linking specific imaging traits with specific gene expression patterns that inform the underlying cellular pathophysiology; imaging features could then serve as molecular surrogates that contribute to the diagnosis, prognosis, and likely gene expression-associated treatment response of various forms of human cancer. This review focuses on high-throughput methods such as microarray analysis of gene expression, their role in cancer research, and in particular, on novel methods of associating gene expression patterns with radiographic imaging phenotypes, known as "radiogenomics." These findings underline a potential future role of both diagnostic and interventional radiologists in genetic assessment of cancer patients with radiographic imaging studies. PMID- 19303234 TI - Achiral liquid chromatography with circular dichroism detection for the determination of carnitine enantiomers in dietary supplements and pharmaceutical formulations. AB - A simple and enantioselective method for the separation and determination of carnitine enantiomers in dietary supplements and pharmaceutical formulation samples is proposed. This method is based on achiral liquid chromatographic separation of carnitine enantiomers from interferences and direct circular dichroism (CD) detection. The calibration curve of the anisotropy factor (g) versus the enantiomeric excess was linear, with a correlation coefficient (R(2)) of 0.996. The precision evaluated by UV peak area and CD peak area was suitable (RSD <5% in all cases). The usefulness of the proposed method was demonstrated by analysing natural dietary supplements and pharmaceutical formulation samples. This method has the advantages of being rapid and precise, without using an expensive chiral column. The method was suitable for the simultaneous determination of both enantiomers and for assessing the chemical purity of carnitine. PMID- 19303235 TI - Quantitative analysis of valiolamine through pre-column derivatization with phenylisocyanate using high-performance liquid chromatography with UV detection: selection of reagent, identification of derivative and optimization of derivatization conditions. AB - This report describes the improved quantitative determination of valiolamine in a medium for microbial culture using high-performance liquid chromatography with UV detection. Valiolamine aqueous solution was dried, dissolved in dimethyl sulfoxide and derivatization performances of phenylisocyanate (PHI), 1-fluoro-2,4 dinitrobenznene and 1-naphthylisothiocyanate were compared in the presence of triethylamine. The PHI was chosen as the most suitable derivatization reagent and the valiolamine-PHI derivative was identified by thin-layer chromatography and electrospray ionization mass spectrometry. The derivative eluted at 10.5 min on a reverse-phase column using a mobile phase composed of 10% acetonitrile in water containing 0.5 mM sodium octyl sulfate (pH 3.0), at a column flow rate of 1.0 mL/min with UV detection at 240 nm. The optimum conditions for derivatization were a reaction temperature of 30 degrees C, reaction time of 30 min, and PHI concentration higher than 33.6 mM. Calibration curves were linear in the range of 0.99-19.95 microg/mL for the standard solutions and 24.9-99.7 microg/mL for the spiked sample. The proposed method was validated and proven to be selective, accurate and precise and suitable for the quantitative analysis of valiolamine in medium for microbial cultures. PMID- 19303236 TI - One foley, two foley, red foley, blue foley. PMID- 19303237 TI - Evaluation and management of moderate to severe pediatric head trauma. AB - A case of pediatric head trauma is presented with a detailed discussion of current concepts in evaluation and treatment. Management of the moderate to severe head-injured child is reviewed, and best practices for emergency department treatment are discussed. BACKGROUND: Pediatric head trauma is a common and potentially devastating injury. Thorough knowledge of the clinical evaluation and treatment will assist the emergency physician in providing optimal care. DISCUSSION: Using a case-based scenario, the initial management strategies along with rationale evidence-based treatments are reviewed. CONCLUSIONS: Computed tomography scan is the diagnostic test of choice for the moderate to severe head injured pediatric patient. Several unique scales to describe and prognosticate the head injury are discussed, although currently, the Glasgow Coma Scale is still the most commonly accepted one. Similar to the adult patient, avoidance of hypotension and hypoxia are key to decreasing mortality. Etomidate and succinylcholine remain the choice of medications for intubation. Hyperventilation should be avoided. PMID- 19303238 TI - Ultrasound-guided peripheral venous access vs. the external jugular vein as the initial approach to the patient with difficult vascular access. AB - BACKGROUND: Traditionally, Emergency Physicians (EPs) have used the external jugular (EJ) vein to gain vascular access in patients who have failed nursing attempts at peripheral access. Recently, some EPs have used ultrasound (USIV) to gain peripheral access. STUDY OBJECTIVE: This study seeks to determine which initial approach by EPs would lead to greater success. METHODS: This was a prospective, randomized study of all adult patients who presented to the Emergency Department (ED) between June and December 2007. Inclusion criteria were failed nursing attempts at peripheral access (at least three). EPs were 2(nd)- or 3(rd)-year residents who had previously performed more than five each of EJs and USIVs. Patients were randomized into either an initial EJ or USIV approach. RESULTS: Sixty patients were enrolled, 32 in the ultrasound group, 28 in the EJ group. Fifteen different EPs performed access. Initial Success: USIV 84% (95% confidence interval [CI] 68-93%) vs. EJ 50% (95% CI 33-67%), p = 0.006. Success if EJ visible: USIV 84% vs. EJ 66% (p = 0.18). Overall success, including data from the crossover pathway: a total of 41 lines were successfully placed by ultrasound out of 46 attempts (89%) vs. 18 out of 33 for EJ (55%), p = 0.001. In total, 59/60 patients (98%) had a peripheral i.v. successfully placed. The percentage of functioning lines when the patient left the ED was: USIV 89% (95% CI 72-96%) vs. EJ 93% (95% CI 68-98%), p = 0.88. CONCLUSION: As an initial approach to all patients with difficult venous access, ultrasound-guided peripheral lines are superior to the EJ approach. However if the EJ was visible, there was no difference in success among the initial approaches. Both techniques, when used together, could achieve peripheral vascular access in 98% of difficult access patients. PMID- 19303239 TI - Unified hands technique for mandibular dislocation. AB - BACKGROUND: The conventional approach to mandibular dislocation involves intraoral bilateral inferior-posterior downward force on the patient's molar occlusal surfaces. DISCUSSION/TECHNIQUE: The unified hands technique is a new method for reducing a mandibular dislocation. The technique involves intraoral unilateral application of both hands on the side of the dislocated mandible. This maneuver was successful in reducing a mandibular dislocation refractory to the traditional method of intraoral bilateral inferior-posterior downward force on the patient's molar occlusal surface. CONCLUSION: The technique described may be useful for the emergent reduction of mandibular dislocations when traditional methods fail. PMID- 19303240 TI - Hypothermia: an unusual indication for gastric lavage. AB - BACKGROUND: Previous reports suggest that gastric lavage holds many risks and is not routinely indicated for decontamination of the overdose patient. OBJECTIVE: To present a case of overdose with concurrent accidental hypothermia where gastric decontamination was utilized. CASE REPORT: A 50-year-old hypothermic, comatose patient was transported to the Emergency Department with a concurrent, massive medication ingestion diagnosed incidentally on a routine abdominal computed tomography scan. Both active and passive rewarming measures, in conjunction with gastric lavage and retrieval of multiple pill fragments, were performed, and the patient survived to hospital discharge without sequelae. Interestingly, the patient admitted to an intentional ingestion of both labetalol and lorazepam. CONCLUSION: Due to hypothermia-mediated changes in metabolism, including gastric atony and decreased hepatic metabolism, gastric lavage may provide additional benefit in the management of severely hypothermic patients with potentially lethal, massive pill ingestions. PMID- 19303241 TI - Ultrasound-guided peripheral intravenous access in the emergency department using a modified Seldinger technique. AB - BACKGROUND: The utility of ultrasound-guided peripheral intravenous access (USGPIV) has been well described. However, few studies have investigated USGPIV techniques. OBJECTIVES: To describe a modified Seldinger technique for USGPIV. METHODS: Emergency Department patients with difficult i.v. access (three or more failed landmark attempts) were prospectively enrolled. USGPIV was attempted using modified Seldinger technique. A 20-gauge, 3.81-cm catheter with integral wire was used for all procedures. The basilic vein was identified using a high-frequency linear probe (5-10 MHz). The needle was inserted into the vein with dynamic guidance in short axis, and the cannula was advanced over a wire. Time from skin puncture to catheter insertion, number of needle sticks, and overall procedure time were recorded. RESULTS: Twenty-five patients were enrolled and underwent USGPIV; success rate was 96% (24/25). The mean number of needle sticks was 1.32 (95% confidence interval 1.12-1.52). Median time from skin to catheter insertion was 68 s (+/- SD 70.5 s). Median total procedure time was 7 min (420 s) (+/- SD 5.23 min). CONCLUSIONS: Modified Seldinger technique is an effective method of USGPIV and is worthy of a prospective comparison with non-Seldinger technique. PMID- 19303242 TI - Maternal smoking, alcohol, and coffee use during pregnancy and son's risk of testicular cancer. AB - It has been suggested that increased risk for testicular cancer occurring worldwide may be due to exposures during fetal development. Lifestyle or environmental exposures may be the most important predictors of risk. However, few studies have directly examined these exposures prospectively. The Child Health and Development Studies is a 40-year follow-up of 20,530 pregnancies occurring between 1959 and 1967. There were 20 cases of testicular cancer diagnosed through 2003 among sons with a maternal interview in early pregnancy. Cases were matched to three controls on birth year and race. Odds ratios and 95% confidence intervals were calculated with exact conditional logistic regression. Compared to controls, mothers of testicular cancer cases were more likely to drink alcohol (unadjusted odds ratio, 3.2; 95% confidence interval, 0.83-15.48 for above vs. below the median for controls) and less likely to drink coffee (unadjusted odds ratio, 0.19; 95% confidence interval, 0.02-1.02 for above vs. below the median). Case mothers were neither more nor less likely to smoke. Although low power may limit interpretation of negative results, the prospective design minimizes bias. In this cohort, maternal serum testosterone in pregnancy was previously reported to be lower in women who drank alcohol. Because populations with high testicular cancer risk also have lower maternal testosterone, we suggest that testosterone could play a role in explaining the higher risk of son's testicular cancer among mothers who drank alcohol during pregnancy. PMID- 19303243 TI - Anastomotic leakage as a risk factor for the long-term outcome after curative resection of colon cancer. AB - AIMS: Long-term outcome for curative colon cancer surgery may be impaired by anastomotic leakage, but most studies regard colon and rectal cancer patients as one group. The aim of this study was to determine whether anastomotic leakage following potentially curative resection for colon cancer is a risk factor for postoperative mortality and for long-term survival. PATIENTS AND METHODS: Medical records of a cohort of 440 consecutive patients undergoing 445 curative resections for explicit colon cancer with primary anastomosis above the peritoneal reflection were reviewed. Therefore patients with rectal cancer were not included. Diagnosis of leakage was made by clinical features or abdominal CT scans. RESULTS: The study population consisted of 266 men and the mean age was 68.6 years. Median follow-up time was 66.5 months. Anastomotic leakage occurred in 12 patients. Four of these died within 30 days after surgery compared to 15 of the remaining 428 patients without leakage (p<0.001). The 5-year overall survival rate was 25% in patients with anastomotic leakage compared to 61.2% in those without leakage (p<0.001). Excluding 30-day mortality, respective values were 33.3 and 63.7% (p=0.02). CONCLUSION: Although anastomotic failure after colon cancer surgery is rare, it is a very severe complication that not only impairs the perioperative morbidity and mortality but also significantly influences the long-term outcome negatively. PMID- 19303244 TI - [Malmignatte (black widow spider) envenomation in France: latrodectism]. PMID- 19303245 TI - [Nerve blocks of the trunk: indications, techniques, advantages and complications]. PMID- 19303246 TI - [Cellular metabolism, temperature and brain injury]. AB - Brain temperature is strongly linked to brain metabolic rate. In the brain, energy metabolism is mainly oxidative. The oxidative metabolism and heat production are therefore strongly related. In normal conditions, heat production consecutive to brain energy metabolism is counterbalanced by heat loss, by using a complex heat exchange system. After major cerebral injuries as subarachnoid haemorrhage or traumatic brain injury, cerebral temperature can often exceed systemic temperature. Moreover, brain temperature can vary independently to systemic temperature, making difficult the prediction of brain temperature from other central temperatures. Mitochondrial dysfunction is probably the corner stone of these post-injury perturbations of brain temperature. Understanding of this phenomenon remains however not complete. PMID- 19303247 TI - [Acute hepatic failure in sickle cell vaso-occlusive crisis]. PMID- 19303248 TI - [Proceedings of the 15th Meeting of the ORL Anesthesia and Resuscitation Club]. PMID- 19303249 TI - [Failure to recover after anaesthesia for surgery of a liver hydatic cyst assigned to hypernatraemia]. PMID- 19303250 TI - Modified trochanteric slide for complex hip arthroplasty: clinical outcomes and complication rates. AB - The sliding trochanteric osteotomy preserves vastus lateralis continuity with the osteotomized greater trochanter (GT) and the abductors. The modified trochanteric sliding osteotomy (MTSO) also preserves the posterior capsule and external rotators to reduce the risk of dislocations. The purpose of this study was to evaluate our clinical and radiographic results of the MTSO exposure. Eighty-three MTSOs were reviewed. Follow-up range was 12 to 126 months. Seventy osteotomies (84.4%) healed with bony union, 9 (10.8%) had fibrous union, and 4 (4.8%) had nonunion. There was no correlation between the width of the osteotomy, intraoperative fragmentation of the GT, or the type of femoral component and the rate of union. Six (7.2%) patients developed a new abductor lurch. Patients with union of the GT had 2.8% of a lurch, and patients with either fibrous union or nonunion had a 30.7% of a lurch (P < .05). There were 4 (4.8%) postoperative dislocations. The benefits of MTSO have been well described, and this study provides evidence of an acceptably low complication rate. PMID- 19303251 TI - Cementless acetabular fixation with and without screws: analysis of stability and migration. AB - The purpose of this study was to compare initial stability and late migration of 775 cementless acetabular components with and without screw fixation. Screw fixation was used in 509 cups and no screws in 266 cups. Average follow-up in the screw fixation group was 6.32 years (range, 2-10 years) and 6.9 years (range, 2 10 years) in the no-screw group. One component (0.2%, osteolysis) in the screw group and one (0.4%, loss of fixation) in the no-screw group required revision. Osteolytic lesions more than 4 cm(2) were noted in 8 (1.6%) screw fixation cups and 2 (0.75%) no-screw fixation cups. No cups in either cohort had radiographic evidence of migration. Screw fixation did not have a favorable or adverse effect on the outcome of acetabular reconstruction. PMID- 19303252 TI - Early nontraumatic fracture of the polyethylene tibial post in a NexGen LPS-Flex posterior stabilized knee prosthesis. AB - Fracture of a polyethylene tibial post in a knee prosthesis after a total knee arthroplasty (TKA) is an uncommon but severe complication of polyethylene tibial post wear. A fractured tibial post results in an unstable and malfunctioning knee. Revision knee surgery is necessary to restore function of the affected knee. We report a case of a nontraumatic fracture of a tibial polyethylene post that occurred in a 63-year-old man with left-sided hemiparesis 3 years after he had undergone left TKA with a NexGen LPS-Flex total knee prosthesis. To our knowledge, this is the first report of a nontraumatic fracture to the tibial post with anterolateral to posterior breakage after a NexGen LPS-Flex TKA. PMID- 19303253 TI - Hepatocellular carcinomas: correlation between time to peak hepatocellular carcinomas enhancement and time to peak aortic enhancement. AB - PURPOSE: To prospectively assess the relationship between the time to peak enhancement of hepatocellular carcinomas (HCC) and that of the aorta at 64 detector computed tomography (CT). MATERIALS AND METHODS: The study prospectively included 43 patients with known HCC. All underwent abdominal CT imaging by using BodyPerfusion CT model. The CT data acquisition was initiated with a delay of 8 15s from the beginning of the contrast material administered. The time-density curves (TDC) of the HCC and the aorta were drawn. The times to peak enhancement of the HCC and the aorta were recorded and the correlation between the time to peak enhancement of the HCC and that of the aorta was analyzed. RESULTS: There were three tendencies of TDC of the HCC enhancement, only 23.3% of them were similar to that of the aorta. The mean time to peak enhancement of the aorta and the HCC (86.1%) was 23.38 s and 30.04 s, respectively. The time to peak enhancement of most HCC was positively and linearly correlated with the time to peak aortic enhancement (r=0.662, P<0.05). CONCLUSION: The result may potentially allow scan delay optimization at contrast material-enhanced CT image in the detection of HCC according to interindividual variability. PMID- 19303255 TI - Pigmented villonodular synovitis of the temporomandibular joint with intracranial extension: a case and literature review. AB - Pigmented villonodular synovitis (PVNS) is a benign, yet locally aggressive proliferative lesion most commonly found in joints of the long bones; it rarely presents in the temporomandibular joint (TMJ). The authors report a case of PVNS involving the TMJ and review similar cases reported in the English literature. This is the first case of PVNS with long-term follow-up of 11 years to include imaging studies. A 36-year-old male with symptoms suggestive of a temporomandibular disorder (TMD) presented with a progressive preauricular/parotid swelling and restricted mandibular range of motion. Imaging suggested a lesion with an aggressive course due to tissue displacement and destruction proximal to the lesion. A multidisciplinary team performed complete excision of the lesion with immediate reconstruction. PVNS often presents with similar symptoms to a TMD, but must be distinguished from symptomatic TMD as it can be highly destructive. Owing to its aggressive nature and potential for recurrence, complete and early extirpation and long-term follow-up with advanced imaging is indicated. PMID- 19303254 TI - MicroRNA in autoimmunity and autoimmune diseases. AB - MicroRNAs (miRNAs) are small conserved non-coding RNA molecules that post transcriptionally regulate gene expression by targeting the 3' untranslated region (UTR) of specific messenger RNAs (mRNAs) for degradation or translational repression. miRNA-mediated gene regulation is critical for normal cellular functions such as the cell cycle, differentiation, and apoptosis, and as much as one-third of human mRNAs may be miRNA targets. Emerging evidence has demonstrated that miRNAs play a vital role in the regulation of immunological functions and the prevention of autoimmunity. Here we review the many newly discovered roles of miRNA regulation in immune functions and in the development of autoimmunity and autoimmune disease. Specifically, we discuss the involvement of miRNA regulation in innate and adaptive immune responses, immune cell development, T regulatory cell stability and function, and differential miRNA expression in rheumatoid arthritis and systemic lupus erythematosus. PMID- 19303256 TI - The Guyuron retroauricular island flap for eyelid and eye socket reconstruction in children. AB - A complete loss of palpebral tissue can occur following a congenital malformation, tumour resection or traumatic injury. This article presents the authors' clinical experience with upper eyelid reconstruction in children using the Guyuron retroauricular island flap. Five cases of severe eyelid defects in children aged between 5 days and 10 years of age (three patients following enucleation and two presenting upper eyelid coloboma of approximately two-thirds of the upper eyelid surface) were treated using this technique. In all cases an optimal closure of the eyelid fissure was achieved and corneal exposure clinically improved. On average, 15% of the initial flap surface was lost. Only one major complication (40% flap necrosis) was reported in the postoperative period. This reconstructive technique can provide complete eyelid reconstruction leaving an inconspicuous scar and causing limited morbidity at the donor zone. PMID- 19303257 TI - Audiovisual integration of speech is disturbed in schizophrenia: an fMRI study. AB - Speech perception is an essential part of social interaction. Visual information (lip movements, facial expression) may supplement auditory information in particular under inadvertent listening situations. Schizophrenia patients have been shown to have a deficit in integrating articulatory motions with the auditory speech input. The goal of this study was to investigate the neural basis of this deficit in audiovisual speech processing in schizophrenia patients by using fMRI. Disyllabic nouns were presented in congruent (audio matches visual information) and incongruent conditions in a slow event related fMRI design. Schizophrenia patients (n=15) were compared to age and gender matched control participants. The statistical examination was conducted by analysis of variance with main factors: audiovisual congruency and group membership. The patients' brain activity differed from the control group as evidenced by congruency by group interaction effects. The pertinent brain sites were located predominantly in the right hemisphere and comprised the pars opercularis, middle frontal sulcus, and superior temporal gyrus. In addition, we observed interactions bilaterally in the fusiform gyrus and the nucleus accumbens. We suggest that schizophrenia patients' deficits in audiovisual integration during speech perception are due to a dysfunction of the speech motor system in the right hemisphere. Furthermore the results can be also seen as a reflection of reduced lateralization of language functions to the left hemisphere in schizophrenia. PMID- 19303258 TI - Decreased tooth size in schizophrenia. PMID- 19303259 TI - Anti-psoriatic effects of indigo naturalis on the proliferation and differentiation of keratinocytes with indirubin as the active component. AB - BACKGROUND: Indigo naturalis has shown efficacy in treating psoriasis in our previous clinical studies. OBJECTIVES: To investigate the potential effect of indigo naturalis on regulating keratinocyte proliferation and differentiation. METHODS: Skin samples from six patients were analyzed for proliferating cell nuclear antigen (PCNA) and involucrin expression by immunohistochemical staining. In addition, indigo naturalis extracts from 10 to 500 microg/ml were added to cultured keratinocytes and cell viability determined. Real-time RT-PCR, Western blotting analysis and indirect immunofluorescent labeling were used to investigate the messenger (m)RNA and protein expressions of PCNA and involucrin. Finally, high-performance liquid chromatography (HPLC) was used to identify major components of indigo naturalis and their in vitro effects compared. RESULTS: Immunohistochemical results demonstrated decreased PCNA and increased involucrin in psoriatic lesions after indigo naturalis treatment. Cultured keratinocytes decreased after indigo naturalis treatment, while G(0)/G(1) arrest was observed to dose-dependently increase. Staining revealed decreased PCNA-stained nuclei and increased cytosolic involucrin in treated keratinocytes. Decreased PCNA and increased involucrin at both the mRNA and protein levels were confirmed. Both major components, indirubin and indigo, could cause G(0)/G(1) phase arrest; however, only indirubin modulated the expressions of PCNA and involucrin similar to indigo naturalis. CONCLUSIONS: Together, these findings indicate that the anti psoriatic effects of indigo naturalis are mediated, at least in part, by modulating the proliferation and differentiation of keratinocytes, with indirubin as the major active component. PMID- 19303260 TI - Proton magnetic resonance spectroscopy of late-life major depressive disorder. AB - The primary goal of this study was to examine the biochemical abnormalities of late-life major depression by using 3-tesla (3-T) proton magnetic resonance spectroscopy ((1)H-MRS). The antidepressant effects on the biochemical abnormalities were investigated as well. Study participants were 27 elderly patients with major depressive disorders (among which 9 were on antidepressant medication) and 19 comparison elderly subjects. (1)H-MRS spectra were acquired from voxels that were placed in the left frontal white matter, left periventricular white matter, and left basal ganglia. Ratios of N-acetylaspartate (NAA), choline (Cho) and myo-inositol to creatine were calculated. Patients with late-life major depressive disorder had a significantly lower NAA/creatine ratio in the left frontal white matter, and higher Cho/creatine and myo inositol/creatine ratios in the left basal ganglia when compared with the control subjects. The myo-inositol correlated with global cognitive function among the patients. The biochemical abnormalities in late-life major depressive disorder were found on the left side of the frontal white matter and the basal ganglia. Neuron degeneration in the frontal white matter and second messenger system dysfunction or glial dysfunction in the basal ganglia are suggested to be associated with late-life depression. PMID- 19303261 TI - Synthesis of aligned hematite nanoparticles on chitosan-alginate films. AB - Iron oxide nanoparticles are being viewed with interest owing to the great potential they have in the biomedical applications like MRI contrast enhancement, targeted drug delivery, hyperthermia and recently in magnetic separation of cancer cells from the body. Templated synthesis has been considered ideal for synthesis of iron oxide nanoparticles as particles are attracted magnetically, in addition to usual flocculation through van der Waals attraction. Biological templates are attractive owing to their biocompatibility and the attractive porosity and surface chemistry that nature provides. Polysaccharides like chitosan and alginate have been employed in the synthesis of a polyion complex, which provided the active-binding sites for iron(II) ions in solution to bind. The natural organization of chitosan and alginate into a porous film has been exploited to synthesize spherical iron oxide nanoparticles through careful calcination of the iron(II) conjugate film. Our experiments indicate that the formed nanoparticles are highly crystalline, confirm to the hematite structure and have a superparamagnetic response with a low coercivity of 116Oe. Particles thus synthesized were highly monodisperse with hydrodynamic diameter of 1.8 nm. The symmetric porosity of the film translates into the synthesis of well-aligned nanoparticles of iron oxide. Compared to synthesis in solution, the film-assisted synthesis offered a greater degree of control over the particle size distribution pattern, with the chitosan-alginate template providing the needed spatial separation to prevent the aggregation due to magnetostatic coupling. Such hematite nanoparticles can either be used directly or converted to paramagnetic magnetite by reduction. Zeta potential measurements indicate highly stable nanoparticles, which can therefore be conjugated to cationic liposomes carrying drugs and magnetically guided to target sites. PMID- 19303262 TI - Cell outer membrane mimetic modification of a cross-linked chitosan surface to improve its hemocompatibility. AB - A novel strategy has been developed to improve the hemocompatibility of chitosan surface by cell outer membrane mimetic structure able to reduce protein adsorption and cell adhesion. Phosphorylcholine dichloride was synthesized and grafted onto a glutaraldehyde-cross-linked chitosan (CS-GA) film surface to prepare phosphorylcholine-coated CS-GA film (CS-GA-PC) through a heterogeneous reaction process. The spectroscopic and contact angle characterization show that a cell outer membrane mimetic structure was formed on the cross-linked chitosan surface, and the significantly improved hemocompatibility of the modified surface was shown by a suppression of 94% on platelet adhesion, a suppression of 60-70% for bovine plasma fibrinogen and bovine serum albumin adsorptions. These results demonstrated that this cell outer membrane mimetic surface modification with phosphorylcholine dichloride is a promising strategy to improve the hemocompatibility of chitosan. PMID- 19303263 TI - Possible mechanisms of Cyclosporin A ameliorated the ischemic microenvironment and inhibited mitochondria stress in tree shrews' hippocampus. AB - OBJECTIVE: The ischemic brain damage is always accompanied by the significant accumulation of glutamate and calcium ions (Ca(2+)). Our objectives were to observe the effects of glutamate and Ca(2+) overloading in tree shrew's hippocampal microenvironment on mitochondrial stress resulting in cytochrome C release and caspase apoptotic gene activation, and to explore the possible mechanism of Cyclosporin A (CsA) inhibiting mitochondrial stress. METHODS: The thrombotic focal cerebral ischemia was induced by photochemical reaction in tree shrews. The extracellular contents of amino acidic neurotransmitters and Ca(2+) were determined, respectively, with high performance liquid chromatography (HPLC) and atomic absorption spectrophotometry at 4, 24 and 72h after cerebral ischemia. The glutamate-calcium chloride solutions were microperfused into hippocampus by a kind of single-pumped push-pull perfusion (SPPP) system under three-dimensional orientation instrument in tree shrews. At 24h, the expression of cytochrome C was observed in perfused lateral hippocampus by immunochemistry. Also, the hippocampus was removed, then mitochondria and cytoplasmic fragment were divided by low temperature centrifugation and the distribution of cytochrome C was assessed through Western blot. Real time fluorescence polymerase chain reaction was used to evaluate the relative amounts of caspase-3 and caspase-9 mRNA. In the treated group, CsA (40mg/kg) was intravenously injected at 6h after the microperfuse or cerebral ischemia. The glutamate-calcium solutions were perfused into the hippocampus and inspected the above-mentioned items at 24h. Data were compared between the two groups (ischemia group vs. sham group, or ischemia group vs. CsA group). RESULTS: Thrombotic cerebral ischemia led to significant increase in extracellular glutamate and Ca(2+) level of hippocampus (P<0.01). The cerebral ischemia group and the microperfusion group, which cytochrome C immunoreactivity increased and Western blot analysis demonstrated that the cytochrome C content in the mitochondria of hippocampal cells decreased (P<0.01), but the cytochrome C in the cytosol increased (P<0.01). When CsA was intravenously injected at 6h after the microperfusion or cerebral ischemia, the cytochrome C expression weakened and its release was diminished to a lesser extent. By real time PCR, in relation to the control group, the caspase-3 and caspase-9 mRNA was higher in the glutamate calcium chloride solution perfused group. CsA treatment cut down the contents of caspase-3 mRNA and caspase-9 mRNA (P<0.01). CONCLUSIONS: It is a primary factor that glutamate and Ca(2+) accumulate in hippocampal microenvironment, which results in proapoptotic protein cytochrome C release from mitochondria into cytoplasm and caspase cascade activation, and finally mitochondria stress and neuronal secondary injury appear. The neuroprotection of CsA is in relation to inhibiting glutamate receptor overactivation and reducing the Ca(2+) influx, which can decrease cytochrome C release and caspase mRNA transition. PMID- 19303264 TI - [Gastroesophageal reflux in infants: myths and realities]. AB - Gastroesophageal reflux (GER) is a common problem in infants but the distinction between GER and GER disease remains difficult. Clinical manifestations such as vomiting, poor weight gain, respiratory disorders, and apneas do always not correlate with the demonstration of reflux episodes. Premature infants frequently suffer from reflux but correlations with apneas are also poor. Esophagitis is a complication suggested in infants experiencing pain but reflux by itself can induce pain as well. The "gold" diagnosis test is pH recording; however, overlap between normal and abnormal indices is obvious. Impedance measurement demonstrates more reflux episodes but non-acid reflux harm is not established. GER disease is probably self-limited in most infants, although it is impossible to predict whether some of them continue to have GER in adult life. The treatment raises doubts concerning indications and efficacy. Overprescription is frequent in infants with regurgitations. Nonpharmacological treatment - small-volume thickened milk and correct positioning - should be the first-line treatment. Prokinetic drugs have not proved their efficacy. Among anti-acid drugs, proton pump inhibitors are the best choice, but their indications are not very clearly established for infants. On the other hand, considerable variations of their metabolism due to the patients' age and genetic factors can explain variations in therapeutic effects. PMID- 19303265 TI - [Gastritis caused by Helicobacter heilmannii probably transmitted from dog to child]. AB - Gastric infection with Helicobacter heilmannii is rare but is known to be associated with chronic active gastritis, peptic ulcer, and low-grade mucosa associated lymphoid tissue lymphoma in humans. In contrast to H. pylori, various H. heilmannii species colonize the stomachs of domestic animals, which might be a reservoir for transmission to humans. We report the case of a 12-year-old boy presenting with chronic gastritis caused by H. heilmannii. Endoscopic examination reveals H. heilmannii-like infection on biopsy samples in his two pet dogs. Sequencing of the 16S and 23S ribosomal DNA by PCR was used to compare the H. heilmannii-like bacterial samples isolated in the boy and his dogs. These DNA amplification methods suggest that the boy was infected by his pet dogs (zoonosis). Our patient was cured by treatment with proton-pump inhibitor and antibiotics. Endoscopic follow-up of the boy showed a complete cure of gastritis and eradication of the bacterium. PMID- 19303266 TI - [A growth delay in an 8-year-old child]. PMID- 19303267 TI - Dietary patterns, cardiovascular risk factors and C-reactive protein in a healthy Italian population. AB - BACKGROUND AND AIMS: Dietary habits have been associated with cardiovascular disease (CVD) risk factors. This study aimed at evaluating the association of non predefined dietary patterns with CVD risk profile and C-reactive protein (CRP). METHODS AND RESULTS: We analyzed 7646 healthy subjects from the Moli-sani project, an on-going cross-sectional cohort study of men and women aged >or=35, randomly recruited from a general Italian population. The Italian EPIC food frequency questionnaire was used. Food patterns were generated using principal factor analysis (PFA) and reduced rank regression (RRR). Three dietary patterns were identified by PFA. The "Olive Oil and Vegetables" pattern, characterized by high intake of olive oil, vegetables, legumes, soups, fruits and fish, was associated with relatively lower values of glucose, lipids, CRP, blood pressure and individual global CVD risk score. The "Pasta and Meat" pattern, characterized by high intake of pasta, tomato sauce, red meat, animal fats and alcohol, was positively associated with glucose, lipids, CRP and CVD risk score. The "Eggs and Sweets" pattern, characterized by positive loadings of eggs, processed meat, margarines, butter, sugar and sweets, was associated with high values of CRP. The first RRR pattern was similar to the "Pasta and Meat" pattern both in composition and association with CVD risk profile. CONCLUSIONS: In a large healthy Italian population, non-predefined dietary patterns including foods considered to be rather unhealthy, were associated with higher levels of cardiovascular risk factors, CRP and individual global CVD risk, whereas a "prudent-healthy" pattern was associated with lower levels. PMID- 19303268 TI - Impact of diabetes on treatment-induced changes in left ventricular structure and function in hypertensive patients with left ventricular hypertrophy. The LIFE study. AB - BACKGROUND AND AIM: Diabetes is associated with left ventricular hypertrophy (LVH) and impaired systolic function in hypertensive patients, but less is known about its impact on LVH regression and functional improvement during antihypertensive treatment. METHODS AND RESULTS: We performed annual echocardiography in 730 non-diabetic and 93 diabetic patients (aged 55-80 years) with hypertension and electrocardiographic LVH during 4.8-year losartan- or atenolol-based treatment in the Losartan Intervention For Endpoint reduction in hypertension (LIFE) study. Baseline mean blood pressure (BP) and LV mass did not differ between groups. Diabetic patients had higher body mass index and pulse pressure, and lower LV ejection fraction, midwall shortening, stress-corrected midwall shortening, and estimated glomerular filtration rate (all p<0.05), and were more likely to have albuminuria. Despite comparable BP reduction in diabetic and non-diabetic groups during treatment (33/18 vs. 28/16mmHg (ns)), diabetes was associated with higher prevalence of persistent LVH (47 vs. 39%, p<0.05). In multivariate analyses, diabetes independently predicted less LV mass reduction and less improvement in stress-corrected LV midwall shortening (both p<0.01). CONCLUSION: Among hypertensive patients with LVH, diabetes is associated with more residual LVH and less improvement in systolic LV function by echocardiography over 4.8 years of antihypertensive treatment. PMID- 19303269 TI - Morphological evaluation of temporomandibular joint after open and closed treatment of type B diacapsular condylar fractures in sheep. AB - The incidence of diacapsular fracture of the mandibular condyle (DFMC) has increased in recent years. However, the specific modality of treatment which would provide maximum benefit and good results for adult patients of DFMC has been controversial. To evaluate and compare morphological changes of temporomandibular joint (TMJ) after open and closed treatments for DFMC, a sheep model of a type B DFMC was created by osteotomy. Eight sheep models of type B DFMC were randomly divided into 2 groups to receive closed treatment (Group 1) and open reduction and internal fixation (ORIF, Group 2), respectively, and morphological changes of TMJ were scored according to the radiological and anatomical criteria of scoring and compared between the 2 groups. Compared with Group 1 animals treated by the closed method, Group 2 animals treated with ORIF received significantly lower scores for morphological changes 12 weeks after treatment. We conclude that ORIF is more efficient than the closed functional treatment in restoring morphology of TMJ to treat type B DFMC. PMID- 19303270 TI - Purification, characterization, and gene cloning of an alkaline serine protease from a highly virulent strain of the nematode-endoparasitic fungus Hirsutella rhossiliensis. AB - Hirsutella rhossiliensis OWVT-1 has substantial potential as a biocontrol agent against plant-parasitic nematodes. Serine proteases have emerged as a potentially useful factor in the nematode-fungus interactions. When grown in liquid culture with the nematode Panagrellus redivivus as the sole nitrogen source, an extracellular alkaline protease (Hasp) was produced by the OWVT-1. The purified Hasp killed the juveniles of the soybean-cyst nematode (Heterodera glycines) and degraded proteins of the nematode cuticle. The molecular mass of Hasp was estimated to be 33 kDa. The optimum pH and temperature for enzyme activity were pH 9 and 75 degrees C. The amino acid sequence obtained by the N-terminal sequence analysis was applied for the primer design to isolate the Hasp cDNA gene, which consists of 1170 bp open reading frame. Analysis of the cDNA and corresponding genomic sequence revealed that Hasp included four exons (279, 186, 513, and 192 bp) divided by three introns (65, 99, and 93 bp). Southern blotting showed that Hasp was a single-copy gene in the genome. The deduced amino acid sequence was very similar to other serine proteases of endoparasitic and egg parasitic fungi of nematodes and of entomopathogenic fungi but was less similar to the serine proteases of nematode-trapping fungi. In a phylogenetic analysis of the amino acid sequences of serine proteases, the serine protease of H. rhossiliensis OWVT-1 clustered with the serine proteases of parasites of nematode eggs rather than with those of the trapping fungi. PMID- 19303271 TI - Effects of rhinacanthins from Rhinacanthus nasutus on nitric oxide, prostaglandin E2 and tumor necrosis factor-alpha releases using RAW264.7 macrophage cells. AB - Three naphthoquinone derivatives, rhinacanthin-C (1), -D (2) and -N (3) were isolated from the leaves of Rhinacanthus nasutus extract and were tested for anti inflammatory activity. The result indicated that all three compounds possessed very potent anti-inflammatory activity against lipopolysaccharide (LPS)-induced nitric oxide release with IC(50) values of 1.8, 6.2 and 3.0 microM, respectively. In addition, the effects of rhinacanthin-C, -D and -N on LPS induced release of prostaglandin E(2) (PGE(2)) and tumor necrosis factor (TNF-alpha) were also examined. It was found that rhinacanthin-C exhibited the most potent on PGE(2) release with an IC(50) value of 10.4 microM, followed by rhinacanthin-D (IC(50)=14.4 microM) and rhinacanthin-N (IC(50)=52.1 microM), whereas those for TNF-alpha were inactive (IC(50)>100 microM). The mechanisms in transcriptional level of rhinacanthin-C were found to inhibit iNOS and COX-2 gene expressions in LPS-induced NO and PGE(2) releases from RAW264.7 cells in concentration-dependent manners. Regarding active constituents for anti-inflammatory activity of R. nasutus, rhinacanthins are responsible for this effect through the inhibition of NO and PGE(2) releases. The finding may support the traditional use of R. nasutus leaves for treatment of the inflammatory-related diseases. PMID- 19303272 TI - Hypoglycemic and hypolipidemic effects of processed Aloe vera gel in a mouse model of non-insulin-dependent diabetes mellitus. AB - The effects of processed Aloe vera gel (PAG) on the course of established diet induced non-insulin-dependent diabetes mellitus (NIDDM) were studied in C57BL/6J mice. NIDDM was induced in C57BL/6J mice by feeding them a high-fat diet. Mice exhibiting diet-induced obesity (DIO) with blood glucose levels above 180mg/dl were selected to examine the antidiabetic effects of PAG. Oral administration of PAG for 8 weeks reduced circulating blood glucose concentrations to a normal level in these DIO mice. In addition, the administration of PAG significantly decreased plasma insulin. The antidiabetic effects of PAG were also confirmed by intraperitoneal glucose tolerance testing. PAG appeared to lower blood glucose levels by decreasing insulin resistance. The administration of PAG also lowered triacylglyceride levels in liver and plasma. Histological examinations of periepididymal fat pad showed that PAG reduced the average size of adipocytes. These results demonstrate that the oral administration of PAG prevents the progression of NIDDM-related symptoms in high-fat diet-fed mice, and suggest that PAG could be useful for treating NIDDM. PMID- 19303273 TI - A randomized controlled trial to assess the safety and efficacy of silymarin on symptoms, signs and biomarkers of acute hepatitis. AB - PURPOSE: Milk thistle or its purified extract, silymarin (Silybum marianum), is widely used in treating acute or chronic hepatitis. Although silymarin is hepatoprotective in animal experiments and some human hepatotoxic exposures, its efficacy in ameliorating the symptoms of acute clinical hepatitis remains inconclusive. In this study, our purpose was to determine whether silymarin improves symptoms, signs and laboratory test results in patients with acute clinical hepatitis, regardless of etiology. METHODS: This is a randomized, placebo-controlled trial in which participants, treating physicians and data management staff were blinded to treatment group. The study was conducted at two fever hospitals in Tanta and Banha, Egypt where patients with symptoms compatible with acute clinical hepatitis and serum alanine aminotransferase (ALT) levels >2.5 times the upper limit of normal were enrolled. The intervention consisted of three times daily ingestion of either a standard recommended dose of 140 mg of silymarin (Legalon, MADAUS GmbH, Cologne, Germany), or a vitamin placebo for four weeks with an additional four-week follow-up. The primary outcomes were symptoms and signs of acute hepatitis and results of liver function tests on days 2, 4 and 7 and weeks 2, 4, and 8. Side-effects and adverse events were ascertained by self report. RESULTS: From July 2003 through October 2005, 105 eligible patients were enrolled after providing informed consent. No adverse events were noted and both silymarin and placebo were well tolerated. Patients randomized to the silymarin group had quicker resolution of symptoms related to biliary retention: dark urine (p=0.013), jaundice (p=0.02) and scleral icterus (p=0.043). There was a reduction in indirect bilirubin among those assigned to silymarin (p=0.012), but other variables including direct bilirubin, ALT and aspartate aminotransferase (AST) were not significantly reduced. CONCLUSIONS: Patients receiving silymarin had earlier improvement in subjective and clinical markers of biliary excretion. Despite a modest sample size and multiple etiologies for acute clinical hepatitis, our results suggest that standard recommended doses of silymarin are safe and may be potentially effective in improving symptoms of acute clinical hepatitis despite lack of a detectable effect on biomarkers of the underlying hepatocellular inflammatory process. PMID- 19303274 TI - Rhodomyrtone: a new candidate as natural antibacterial drug from Rhodomyrtus tomentosa. AB - Rhodomyrtone [6,8-dihydroxy-2,2,4,4-tetramethyl-7-(3-methyl-1-oxobutyl)-9-(2 methylpropyl)-4,9-dihydro-1H-xanthene-1,3(2H)-di-one] from Rhodomyrtus tomentosa (Aiton) Hassk. displayed significant antibacterial activities against gram positive bacteria including Bacillus cereus, Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Staphylococcus epidermidis, Streptococcus gordonii, Streptococcus mutans, Streptococcus pneumoniae, Streptococcus pyogenes, and Streptococcus salivarius. Especially noteworthy was the activity against MRSA with a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) ranging from 0.39 to 0.78 microg/ml. As shown for S. pyogenes, no surviving cells were detected within 5 and 6h after treatment with the compound at 8MBC and 4MBC concentrations, respectively. Rhodomyrtone displays no bacteriolytic activity, as determined by measurement of the optical density at 620 nm. A rhodomyrtone killing test with S. mutans using phase contrast microscopy showed that this compound caused a few morphological changes as the treated cells were slightly changed in color and bigger than the control when they were killed. Taken together, the results support the view that rhodomyrtone has a strong bactericidal activity on gram-positive bacteria, including major pathogens. PMID- 19303275 TI - 4-Methoxycinnamaldehyde inhibited human respiratory syncytial virus in a human larynx carcinoma cell line. AB - 4-Methoxycinnamaldehyde, an active constituent of Agastache rugosa, was examined for its cytoprotective activity against RSV by XTT method in human larynx carcinoma cell line. 4-Methoxycinnamaldehyde could effectively inhibit cytopathic effect of RSV (p<0.0001) with an estimated IC(50) of 0.055microg/ml and a selectivity index (SI) of 898.2. 4-Methoxycinnamaldehyde (0.03microg/ml) could inhibit viral entrance by interfering viral attachment (IC(50) of 0.06microg/ml; p<0.0001) and internalization (IC(50) of 0.01microg/ml; p<0.0001). 4 Methoxycinnamaldehyde significantly increased the basal production of IFN (p=0.0015), but not the virus-induced IFN production. Therefore, its cytoprotective activity against RSV was not mediated by interferon. In conclusion, 4-methoxycinnamaldehyde might be helpful to manage the disease induced by RSV infection. PMID- 19303276 TI - Kaempferol from the leaves of Apocynum venetum possesses anxiolytic activities in the elevated plus maze test in mice. AB - The present work evaluated the anxiolytic activity of an aqueous extract of Apocynum venetum L. (Apocynaceae) and bioguided its fractionation using the elevated plus maze (EPM) in mice as a model of anxiety. A single treatment of AV extract markedly increased the percentage time spent on the open arms of the EPM in two distinct concentration ranges of 22.5-30 and 100-125 mg/kg p.o., respectively, indicating a putative anxiolytic-like activity. Fractions showing anxiolytic effects in concentrations equal to 30 or 125 mg/kg of whole extract were antagonized using the benzodiazepine antagonist flumazenil (3 mg/kg i.p.) or the 5-HT(1A) receptor antagonist WAY-100635 (0.5 mg/kg i.p.). All active fractions in a concentration equal to 125 mg/kg were effectively blocked by the benzodiazepine antagonist flumazenil, while the anxiolytic activities of fractions in the lower dose equivalent to 30 mg/kg of whole extract were inhibited by the 5-HT(1A) receptor antagonist WAY-100635. Through further separation of AV fractions it was possible to isolate and characterize the flavonol kaempferol which showed an anxiolytic-like activity in concentrations from 0.02 to 1.0 mg/kg p.o. The anxiolytic activity of kaempferol was partially antagonized by concomitant administration of flumazenil, but not by WAY-100635. In conclusion, our study clearly demonstrates that AV extract possesses anxiolytic-like activity and that at least one of its flavonoids, kaempferol, can elicit the same kind of neuropharmacological activity. PMID- 19303277 TI - Fetal learning and memory: weak associations with the early essential polyunsaturated fatty acid status. AB - To study the potential associations between fetal brain functions and the early essential polyunsaturated fatty acid (ePUFA) status, fetal learning and memory were assessed by repeated habituation rate measurements (HR) in fetuses of 30, 32, 34 or 36 weeks gestational age (GA). HR tests were repeated 10 min later. Both measurements were replicated in a second session at GA 38. Fetal short-term memory (STM) and long-term memory (LTM) were calculated from these habituation rates and related to concentrations of ePUFAs and their status markers, measured in umbilical artery wall phospholipids. The only relevant associations observed were positive trends (0.01095% removal) combined with a low effluent sBOD(5) concentration (<5mg/L) was consistently achieved. Moreover, phosphate concentrations in the effluent were maintained below 7.5 mg P/L (>81% reduction). This study demonstrates the potential to combine a digester and a denitrification filter in a single unit to efficiently remove nitrate and phosphate from hydroponic wastewater in a single unit. PMID- 19303288 TI - Discovery of potent non-urea inhibitors of soluble epoxide hydrolase. AB - Soluble epoxide hydrolase (sEH) is a novel target for the treatment of hypertension and vascular inflammation. A new class of potent non-urea sEH inhibitors was identified via high throughput screening (HTS) and chemical modification. IC(50)s of the most potent compounds range from micromolar to low nanomolar. PMID- 19303289 TI - Discovery of oxazolo[4,5-b]pyridines and related heterocyclic analogs as novel SIRT1 activators. AB - SIRT1 is an NAD(+)-dependent protein deacetylase that appears to produce beneficial effects on metabolic parameters such as glucose and insulin homeostasis. Activation of SIRT1 by resveratrol (1) has been shown to modulate insulin resistance, increase mitochondrial content and prolong survival in lower organisms and in mice on a high fat diet. Herein, we describe the identification and SAR of a series of oxazolo[4,5-b]pyridines as novel small molecule activators of SIRT1 which are structurally unrelated to and more potent than resveratrol. PMID- 19303290 TI - Discovery and structure-activity study of a novel benzoxaborole anti-inflammatory agent (AN2728) for the potential topical treatment of psoriasis and atopic dermatitis. AB - A series of phenoxy benzoxaboroles were synthesized and screened for their inhibitory activity against PDE4 and cytokine release. 5-(4-Cyanophenoxy)-2,3 dihydro-1-hydroxy-2,1-benzoxaborole (AN2728) showed potent activity both in vitro and in vivo. This compound is now in clinical development for the topical treatment of psoriasis and being pursued for the topical treatment of atopic dermatitis. PMID- 19303291 TI - 5,6-Diarylanthranilo-1,3-dinitriles as a new class of antihyperglycemic agents. AB - Various functionalized mono- and diarylanthranilo-1,3-dinitriles were synthesized and evaluated for their in vitro antihyperglycemic activity against the PTP-1B, glucose-6-phosphatase, glycogen phosphorylase and alpha-glucosidase enzymes. Among various screened compounds, 5,6-diaryl substituted anthranilo-1,3 dinitriles 3a, 3b, and 3d showed good inhibitory activity against PTP-1B with IC(50) values of 58-72 microM. Three of the test compounds showed significant (25 37%) lowering of plasma glucose level at 24h in sucrose-challenged streptozotocin induced diabetic Sprague-Dawley rat model. PMID- 19303292 TI - Efficient synthesis of oligonucleotide conjugates on solid-support using an (aminoethoxycarbonyl)aminohexyl group for 5'-terminal modification. AB - Solid-support conjugation at the 5'-terminal primary amine of oligonucleotides is a convenient and powerful method for introducing various functional groups. However, conventional aliphatic amines do not necessarily provide conjugates with sufficient yields. To improve the modification efficacy, we used the amino-linker (aminoethoxycarbonyl)aminohexyl group (ssH-linker), for solid-support conjugation. In the ssH-linker terminal modification, reactive free amino group could be easily presented onto a solid-support due to rapid removal of the amino protecting group, and activated amino acids or cholesterol molecules could be covalently connected more efficiently than to typical 6-aminohexyl-linkers. Based on these results, the ssH-linker can be a useful terminal modification for the solid-support conjugation of functional molecules. PMID- 19303293 TI - Dendrimers ended by non-symmetrical azadiphosphonate groups: synthesis and immunological properties. AB - The synthesis and characterization of new series of phosphorus-containing dendrimers ended by non-symmetrical azamonophosphonates, or azadiphosphonates, or azadiphosphonic acid salts are reported. The sodium salts of the non-symmetrical azadiphosphonic dendrimers are soluble in water. Their influence towards human immune blood cells is assayed ex vivo. PMID- 19303294 TI - Late-onset axial myopathy with cores due to a novel heterozygous dominant mutation in the skeletal muscle ryanodine receptor (RYR1) gene. AB - Mutations in the skeletal muscle ryanodine receptor (RYR1) gene have been associated with a wide range of phenotypes including the malignant hyperthermia (MH) susceptibility trait, Central Core Disease (CCD) and other congenital myopathies characterized by early onset and predominant proximal weakness. We report a patient presenting at 77 years with a predominant axial myopathy associated with prominent involvement of spine extensors, confirmed on MRI and muscle biopsy, compatible with a core myopathy. RYR1 mutational analysis revealed a novel heterozygous missense mutation (c.119G>T; p.Gly40Val) affecting the RYR1 N-terminus, previously predominantly associated with MH susceptibility. This case expands the spectrum of RYR1-related phenotypes and suggests that MH-related RYR1 mutations may give rise to overt neuromuscular symptoms later in life, with clinical features not typically found in CCD due to C-terminal hotspot mutations. Late-onset congenital myopathies may be under-recognised and diagnosis requires a high degree of clinical suspicion. PMID- 19303295 TI - Use of SNP array analysis to identify a novel TRIM32 mutation in limb-girdle muscular dystrophy type 2H. AB - Molecular diagnosis of monogenic diseases with high genetic heterogeneity is usually challenging. In the case of limb-girdle muscular dystrophy, multiplex Western blot analysis is a very useful initial step, but that often fails to identify the primarily affected protein. We report how homozygosity analysis using a genome-wide SNP array allowed us to solve the diagnostic enigma in a patient with a moderate form of LGMD, born from consanguineous parents. The genome-wide scan performed on the patient's DNA revealed several regions of homozygosity, that were compared to the location of known LGMD genes. One such region indeed contained the TRIM32 gene. This gene was previously found mutated in families with limb-girdle muscular dystrophy type 2H (LGMD2H), a mild autosomal recessive myopathy described in Hutterite populations and in 4 patients with a diagnosis of sarcotubular myopathy. A single missense mutation was found in all these patients, located in a conserved domain of the C-terminal part of the protein. Another missense mutation affecting the N-terminal part of TRIM32, observed in a single consanguineous Bedouin family, was reported to cause the phenotypically unrelated and genetically heterogeneous Bardet-Biedl syndrome, defining the BBS11 locus. Sequencing of TRIM32 in our patient revealed a distal frameshift mutation, c.1753_1766dup14 (p.Ile590Leu fsX38). Together with two recently reported mutations, this novel mutation confirms that integrity of the C terminal domain of TRIM32 is necessary for muscle maintenance. PMID- 19303296 TI - Recent region-wide declines in Caribbean reef fish abundance. AB - Profound ecological changes are occurring on coral reefs throughout the tropics, with marked coral cover losses and concomitant algal increases, particularly in the Caribbean region. Historical declines in the abundance of large Caribbean reef fishes likely reflect centuries of overexploitation. However, effects of drastic recent degradation of reef habitats on reef fish assemblages have yet to be established. By using meta-analysis, we analyzed time series of reef fish density obtained from 48 studies that include 318 reefs across the Caribbean and span the time period 1955-2007. Our analyses show that overall reef fish density has been declining significantly for more than a decade, at rates that are consistent across all subregions of the Caribbean basin (2.7% to 6.0% loss per year) and in three of six trophic groups. Changes in fish density over the past half-century are modest relative to concurrent changes in benthic cover on Caribbean reefs. However, the recent significant decline in overall fish abundance and its consistency across several trophic groups and among both fished and nonfished species indicate that Caribbean fishes have begun to respond negatively to habitat degradation. PMID- 19303297 TI - A hypervariable invertebrate allodeterminant. AB - Colonial marine invertebrates, such as sponges, corals, bryozoans, and ascidians, often live in densely populated communities where they encounter other members of their species as they grow over their substratum. Such encounters typically lead to a natural histocompatibility response in which colonies either fuse to become a single, chimeric colony or reject and aggressively compete for space. These allorecognition phenomena mediate intraspecific competition, support allotypic diversity, control the level at which selection acts, and resemble allogeneic interactions in pregnancy and transplantation. Despite the ubiquity of allorecognition in colonial phyla, however, its molecular basis has not been identified beyond what is currently known about histocompatibility in vertebrates and protochordates. We positionally cloned an allorecognition gene by using inbred strains of the cnidarian, Hydractinia symbiolongicarpus, which is a model system for the study of invertebrate allorecognition. The gene identified encodes a putative transmembrane receptor expressed in all tissues capable of allorecognition that is highly polymorphic and predicts allorecognition responses in laboratory and field-derived strains. This study reveals that a previously undescribed hypervariable molecule bearing three extracellular domains with greatest sequence similarity to the immunoglobulin superfamily is an allodeterminant in a lower metazoan. PMID- 19303298 TI - Cdk1 negatively regulates midzone localization of the mitotic kinesin Mklp2 and the chromosomal passenger complex. AB - The survival of eukaryotes depends on the accurate coordination of mitosis with cytokinesis. Key for the coordination of both processes is the chromosomal passenger complex (CPC) comprising Aurora-B, INCENP, survivin, and borealin. The translocation of the CPC from centromeres to the spindle midzone, a structure composed of antiparallel microtubules, at anaphase onset is critical for the completion of cytokinesis. In mammalian cells, the mitotic kinesin Mklp2 is essential for recruitment of the CPC to the spindle midzone. However, the mechanism regulating the binding of Mklp2 to microtubules has remained unknown. Here, we demonstrate that Mklp2 and the CPC mutually depend on each other for midzone localization; i.e., Mklp2 is mislocalized in INCENP-RNAi cells and vice versa. Remarkably, INCENP is required for localization of Mklp2 to the ends of stable microtubules in cells with low Cdk1 activity. In vitro assays revealed that the association between the CPC and Mklp2 is negatively regulated by Cdk1. Collectively, our data suggest that anaphase onset triggers the association between the CPC and Mklp2 and that this association targets the CPC-Mklp2 complex to the ends of stable microtubules in the spindle midzone. PMID- 19303299 TI - Peripheral visual circuits functionally segregate motion and phototaxis behaviors in the fly. AB - Like the mammalian visual cortex, the fly visual system is organized into retinotopic columns. A widely accepted biophysical model for computing visual motion, the elementary motion detector proposed nearly 50 years ago posits a temporal correlation of spatially separated visual inputs implemented across neighboring retinotopic visual columns. Whereas the inputs are defined, the neural substrate for motion computation remains enigmatic. Indeed, it is not known where in the visual processing hierarchy the computation occurs. Here, we combine genetic manipulations with a novel high-throughput dynamic behavioral analysis system to dissect visual circuits required for directional optomotor responses. An enhancer trap screen of synapse-inactivated neural circuits revealed one particularly striking phenotype, which is completely insensitive to motion yet displays fully intact fast phototaxis, indicating that these animals are generally capable of seeing and walking but are unable to respond to motion stimuli. The enhancer circuit is localized within the first optic relay and strongly labels the only columnar interneuron known to interact with neighboring columns both in the lamina and medulla, spatial synaptic interactions that correspond with the two dominant axes of elementary motion detectors on the retinal lattice. PMID- 19303300 TI - Universal recognition of three basic emotions in music. AB - It has long been debated which aspects of music perception are universal and which are developed only after exposure to a specific musical culture. Here, we report a crosscultural study with participants from a native African population (Mafa) and Western participants, with both groups being naive to the music of the other respective culture. Experiment 1 investigated the ability to recognize three basic emotions (happy, sad, scared/fearful) expressed in Western music. Results show that the Mafas recognized happy, sad, and scared/fearful Western music excerpts above chance, indicating that the expression of these basic emotions in Western music can be recognized universally. Experiment 2 examined how a spectral manipulation of original, naturalistic music affects the perceived pleasantness of music in Western as well as in Mafa listeners. The spectral manipulation modified, among other factors, the sensory dissonance of the music. The data show that both groups preferred original Western music and also original Mafa music over their spectrally manipulated versions. It is likely that the sensory dissonance produced by the spectral manipulation was at least partly responsible for this effect, suggesting that consonance and permanent sensory dissonance universally influence the perceived pleasantness of music. PMID- 19303301 TI - Local cues and asymmetric cell divisions underpin body plan transitions in the moss Physcomitrella patens. AB - BACKGROUND: Land plants evolved from aquatic algae more than 450 million years ago. Algal sisters of land plants grow through the activity of apical initial cells that cleave either in one plane to generate filaments or in two planes to generate mats. Acquisition of the capacity for cell cleavage in three planes facilitated the formation of upright bushy body plans and enabled the invasion of land. Evolutionary transitions between filamentous, planar, and bushy growth are mimicked within moss life cycles. RESULTS: We have developed lineage analysis techniques to assess how transitions between growth forms occur in the moss Physcomitrella patens. We show that initial cells giving rise either to new filaments or bushy shoots are frequently juxtaposed on a single parent filament, suggesting a role for short-range cues in specifying differences in cell fate. Shoot initials cleave four times to establish a tetrahedral shape and subsequently cleave in three planes, generating bushy growth. Asymmetric and self replacing divisions from the tetrahedral initial generate leaf initials that divide asymmetrically to self-replace and to produce daughter cells with restricted fate. The cessation of division in the leaf is distributed unevenly and contributes to final leaf shape. CONCLUSIONS: In contrast to flowering plants, changes in body plan in P. patens are regulated by cues acting at the level of single cells and are mediated through asymmetric divisions. Genetic mechanisms regulating shoot and leaf development in P. patens are therefore likely to differ substantially from mechanisms operating in plants with more recent evolutionary origins. PMID- 19303302 TI - Microsurgical treatment and functional outcomes of multi-segment intramedullary spinal cord tumors. AB - We aimed to prospectively analyze correlations between clinical features and histological classification of multi-segment intramedullary spinal cord tumors (MSICTs), and the extent of microsurgical resection and functional outcomes. Fifty-six patients with MSICTs underwent microsurgery for tumor removal using a posterior approach. The tumor was exposed through a dorsal myelotomy. Pre operative and post-operative nervous function was scored using the Improved Japanese Orthopaedic Association (IJOA) grading system. Correlation analyses were performed between functional outcome (IJOA score) and histological features, age, tumor location, and the longitudinal extent of spinal cord involvement. The most frequently involved levels were the medullo cervical and the cervicothoracic regions (51.8%, 29/56) followed by the conus terminalis (26.8%, 15/56) and the thoracic region (14.3%, 8/56). Ependymoma was the most frequent MSICT type, seen in 22 of 56 patients (39%), followed by low grade astrocytoma (17 patients, 30%) and glioblastoma multiforme (3 patients, 5%). Gross total tumor removal was achieved in 33 cases (58%), subtotal resection in 4 (7%), and partial resection in 16 (28%). The histological classification of the tumor was the most important factor influencing the extent of surgical removal (chi2=22.17, p=0.00). The overall difference between pre-operative and post-operative neurological state was not significant (chi2=5.44, p=0.61). Thus, MSICTs were most commonly seen in the medullo cervical and cervicothoracic regions, with ependymoma and low grade astrocytoma the most common tumour types. We stress the importance of early microsurgical treatment for MSICTs while the patients do not have severe dysfunction. PMID- 19303303 TI - Assessment of the corona radiata sensory tract using awake surgery and tractography. AB - Anatomical localization of brain function can be achieved by functional changes during awake surgery combined with tractography constructed by diffusion tensor imaging studies. We aimed to use these techniques to characterize the sensory tract in the corona radiata in patients with closely associated brain tumors. Of nine patients who had brain tumors in the primary sensory area (S1) and who underwent awake surgery between October 2004 and July 2007, two patients showed deterioration in deep sensation during and after awake surgery. Both of these patients also developed involuntary movements (for patient 1 this was myoclonus of the left hand, while patient 2 experienced unintentional lifting of the arm). In these two patients, tumors were located just beside the sensory tract in the corona radiata of the right hemisphere. In patient 2, Wallerian degeneration of the sensory tract and concomitant deterioration of superficial and deep sensation were observed at 6 months after awake surgery. These results suggest that damage to a closely associated sensory tract in the corona radiata is critical to the development of sensory deficits and involuntary movements. For patients who undergo surgical resection of S1 brain tumors, pre-operative tractography to detect the sensory tract in the corona radiata may allow protection of the sensory tract during awake surgery, thereby preventing post-operative sensory deficits. PMID- 19303304 TI - Voxel-based morphometry in the detection of dysplasia and neoplasia in childhood epilepsy: limitations of grey matter analysis. AB - The purpose of this exploratory investigation was to evaluate voxel-based morphometry (VBM) in detecting lesions underlying childhood epilepsy, and to establish the optimal image processing and statistical parameters in this context. The patients were 16 children (10 boys) aged 5.9 to 15.2 years (mean 11.3 years) with epilepsy and focal cortical dysplasia (FCD) or neoplasia. The control group comprised 24 normal children (12 boys), age matched to the patients. MRI volumes were spatially normalised to a custom template and segmented into grey matter (GM) and white matter. Using statistical parametric mapping, the GM segment from each patient was then contrasted with the mean GM segment of the control group utilising different VBM post-processing methods. Maps showing increased/decreased areas of GM concentration or volume were generated and compared with visually identified lesions. The results indicated that conservative VBM parameters of linear normalisation with no modulation produced the highest rates of lesion detection, which were identical for FCD and neoplasia at 5/8 lesions. These preliminary data suggest that VBM analysis of GM using conservative parameters can usually detect FCD and neoplasia in the MRI of children with epilepsy, but sensitivity may be inadequate for routine clinical application. Further refinement of the technique may be necessary. PMID- 19303305 TI - Co-evolution of primordial membranes and membrane proteins. AB - Studies of the past several decades have provided major insights into the structural organization of biological membranes and mechanisms of many membrane molecular machines. However, the origin(s) of the membrane(s) and membrane proteins remains enigmatic. We discuss different concepts of the origin and early evolution of membranes with a focus on the evolution of the (im)permeability to charged molecules such as proteins, nucleic acids and small ions. Reconstruction of the evolution of F-type and A/V-type membrane ATPases (ATP synthases), which are either proton- or sodium-dependent, might help us to understand not only the origin of membrane bioenergetics but also of membranes themselves. We argue that evolution of biological membranes occurred as a process of co-evolution of lipid bilayers, membrane proteins and membrane bioenergetics. PMID- 19303306 TI - 12-Substituted 2,3-dimethoxy-8,9-methylenedioxybenzo[i]phenanthridines as novel topoisomerase I-targeting antitumor agents. AB - 2,3-Dimethoxy-8,9-methylenedioxybenzo[i]phenanthridine and a few of its 12 substituted analogs are active as TOP1-targeting agents. Studies were performed to further evaluate the potential of this series of non-camptothecin TOP1 targeting agents. The influence of a hydroxymethyl, formyl, N,N dimethylaminomethyl, 2-(N,N-dimethylamino)ethyl, 3-(N,N-dimethylamino)propyl), and 4-(N,N-dimethylamino)butyl substituent at the 12-position on TOP1-targeting activity and tumor cell growth was evaluated. In addition, the relative pharmacologic activities of the 12-carboxamide analog, as well as its N-methyl and N,N-dimethyl derivatives were assessed. PMID- 19303307 TI - Modified low molecular weight cyclic peptides as mimetics of BDNF with improved potency, proteolytic stability and transmembrane passage in vitro. AB - We recently reported the development of the BDNF mimetic peptide cyclo-[dPAKKR] 1 which promotes the survival of cultured sensory neurons via a trkB independent mechanism [Fletcher, J. M.; Morton, C. M.; Zwar, R. A.; Murray, S. S.; O'Leary, P. D.; Hughes, R. A. J. Biol. Chem.2008, 283, 33375]. In the present study we prepared a series of hydrophobically-modified analogues of 1 with an eye to improving its pharmacokinetic properties. Select members of this second generation of compounds showed improved biological activity, stability in plasma, and an ability to cross model biological membranes. PMID- 19303308 TI - Molecular modeling studies and in vitro bioactivity evaluation of a set of novel 5-nitro-heterocyclic derivatives as anti-T. cruzi agents. AB - In this study, in vitro anti-T. cruzi activity assays of nifuroxazide (NX) analogues, such as 5-nitro-2-furfuryliden and 5-nitro-2-theniliden derivatives, were performed. A molecular modeling approach was also carried out to relate the lipophilicity potential (LP) property and biological activity data. The majority of the NX derivatives showed increased anti-T. cruzi activity in comparison to the reference drug, benznidazole (BZN). Additionally, the 5-nitro-2-furfuryliden derivatives presented better pharmacological profile than the 5-nitro-2 theniliden analogues. The LP maps and corresponding ClogP values indicate that there is an optimum lipophilicity value, which must be observed in the design of new potential anti-T. cruzi agents. PMID- 19303309 TI - The identification of novel PLC-gamma inhibitors using virtual high throughput screening. AB - Phospholipase C-gamma (PLC-gamma) has been identified as a possible biological target for anticancer drug therapy but suitable inhibitors are lacking. Therefore, in order to identify active compounds (hits) virtual high throughput screening was performed. The crystal structure of the PLC-delta isoform was used as a model docking scaffold since no crystallographic data are available on its gamma counterpart. A pilot screen was performed using approximately 9.2x10(4) compounds, where the robustness of the methodology was tested. This was followed by the main screening effort where approximately 4.4x10(5) compounds were used. In both cases, plausible compounds were identified (virtual hits) and a selection of these was experimentally tested. The most potent compounds were in the single digit micro-molar range as determined from the biochemical (Flashplate) assay. This translated into approximately 15 microM in a functional assay in cells. About 30% of the virtual hits showed activity against PLC-gamma (IC(50)<50 microM). PMID- 19303310 TI - The use of a neutron backscatter technique for in-situ water measurement in paper recycling industry. AB - A bulk of used paper supplied to recycling industry may contain water in their internal voids. This is because the price of the used paper is currently based on their weight and it has a huge potential of suppliers to add with water in order to increase the price. Currently used methods for detecting moisture content in a paper are restricted to a sheet of paper only. This paper presents a non intrusive method for quick and in-situ measurement of water content in a bulk of used paper. The proposed method extends the capability of common paper moisture gauge, by using a neutron device. A fast neutron source (Am-Be 241) and a portable backscattering neutron detector are used for water measurement. It theoretically indicates that the slow neutron counts can be correlated to the hydrogen or water level in a paper. The method has the potential of being used by the paper-recycling industry for rapid and non-destructive measurement of water in a bulk of used paper. PMID- 19303311 TI - Optimized therapeutic neutron beam for accelerator-based BNCT by analyzing the neutron angular distribution from (7)Li(p,n)(7)Be reaction. AB - Perpendicular neutrons (i.e., solid angle bin of 50-150 degrees ) among ones generated from (7)Li(p,n)(7)Be reaction were used to produce an optimized therapeutic neutron beam for accelerator-based BNCT. A new beam port assembly was also designed to shape the fast neutrons into epithermal ones and to reduce unnecessary radiation including gammas. As a result of a simulation, it is found that a tumor at a depth of 60mm from the head skin could be treated within 5 minutes, if a typical tumor is assumed to be taken about 20RBEGy for therapeutic treatment. It is, thus, expected that the neutrons emitted into the solid angle bin of 50-150 degrees from (7)Li(p,n)(7)Be reaction are very effective in producing epithermal neutron beams for BNCT. PMID- 19303312 TI - Sorption of U(VI) on goethite: effects of pH, ionic strength, phosphate, carbonate and fulvic acid. AB - U(VI) sorption on goethite was investigated as functions of pH, solid-to-liquid ratio (m/V), ionic strength and U(VI) concentration by a batch experimental method. Effects of phosphate, carbonate and fulvic acid (FA) on U(VI) sorption were examined. It was found that the sorption of U(VI) increases from 0% to 100% over the pH range of 2.5-4.5 and keeps constant in the high pH range. The sorption of U(VI) on goethite is insensitive to ionic strength. Different surface complexes in the framework of double-layer model were examined for fitting the sorption of U(VI) on goethite. A model with two mononuclear inner-sphere surface complexes, identical with triple bond SOUO(2)(+) and identical with triple bond SOUO(2)OH, was found capable of reproducing the pH sorption edges, the sorption isotherms and the sorption data with variable m/V in this study. The proposed model can also interpret the pH sorption edge collected at P(CO2)=10(-3.58) atm without considering any ternary surface complexes of carbonate. Moreover, it was found that the presence of phosphate at relatively high concentration (6 x 10(-4) mol/L) promotes U(VI) sorption. The presence of FA of 20 mg/L has little effect on the sorption of U(VI) on goethite. PMID- 19303313 TI - Production of Sn-117m in the BR2 high-flux reactor. AB - The BR2 reactor is a 100MW(th) high-flux 'materials testing reactor', which produces a wide range of radioisotopes for various applications in nuclear medicine and industry. Tin-117m ((117m)Sn), a promising radionuclide for therapeutic applications, and its production have been validated in the BR2 reactor. In contrast to therapeutic beta emitters, (117m)Sn decays via isomeric transition with the emission of monoenergetic conversion electrons which are effective for metastatic bone pain palliation and radiosynovectomy with lesser damage to the bone marrow and the healthy tissues. Furthermore, the emitted gamma photons are ideal for imaging and dosimetry. PMID- 19303314 TI - Development of a beta gauge system for a fabric density measurement. AB - A beta gauge system was designed and fabricated for a fabric density measurement with high sensitivity in real time. An ionization chamber and 3.7GBq (85)Kr was chosen as a beta radiation sensor and a beta source. An ionization chamber, which had a 0.5microm-thick aluminized mylar film as an incident window, was constructed with a guard electrode structure in consideration of minimizing leakage currents. Xe gas was chosen as a filling gas to increase sensitivity of an ionization chamber. Linearity of the ionization currents against the densities was 0.9894 in the selected measurement range. And the deviation of the ionization currents at a standard density was below 0.02%. The ranges of the density measurement and the active area were from 20 to 999g/cm(2) with +/-1% accuracy and 40mm diameters. The fabricated beta gauge system can be applied to fields such as in the paper, film, and plastic production industries. PMID- 19303315 TI - Underivatized polyamine analysis in plant samples by ion pair LC coupled with electrospray tandem mass spectrometry. AB - Polyamines are key regulators of cell development and many plant responses to environmental challenges, however, their functions still remain unclear in complex interactions with other hormones and in biotic or abiotic stress. This lack of knowledge derives from the difficulties on measuring natural polyamines in plants. Here, we present a fast multiresidue method for putrescine (Put), 1,3 diaminopropane (DAP), l-ornithine, spermidine (Spd) and spermine (Spn) measurements in plant samples. Polyamine determination is based on a perchloric acid extraction followed by a simple filtration procedure without previous derivatization. Polyamines are resolved by HPLC in a C18 common column and quantified by electrospray ionization tandem mass spectrometry. (13)C(4) putrescine and 1,7-diaminoheptane standards were added prior to sample extraction to achieve an accurate quantification in a single run. Chromatography of polyamines presents poor retention when reverse phase C18 common columns are used, because they are very polar compounds and contain several positive charges. To circumvent this problem ionic pairing technique has been used successfully with heptafluorobutyric acid (HFBA) at 1mM in the aqueous phase and 25mM in the sample. Improvement of the signal depleted by HFBA has been achieved by adding 1% of propionic acid to the aqueous and organic eluents. All together, gives a method accurate enough to determine polyamines in plants. To demonstrate the usefulness of the method it has been validated in Arabidopsis thaliana samples and polyamines have been determined in several genotypes that over express (35S::ADC2 line 3.6) or are disrupted (adc2) in the Arginine Decarboxylase2 (ADC2) gene. PMID- 19303316 TI - Prophylactic aortic arch debranching during type A aortic dissection repair. PMID- 19303317 TI - Variability in photobleaching yields and their related impacts on optical conditions in subtropical lakes. AB - Changes in the concentration and spectral absorption of chromophoric dissolved organic matter (CDOM) may strongly condition the optical properties of tropical and subtropical water bodies. We examined the spatial distribution of CDOM related absorption, spectral slope and vertical attenuation of solar radiation in two shallow lakes in the Esteros del Ibera wetland system. In situ measurements were made to examine spatial variations in photobleaching yields in natural lake conditions. The results showed that "fresh" allochthonous CDOM is more susceptible to phototransformations than either "aged" allochthonous organic matter or autochthonous sources, if the distances from sources are considered as proxies for residence time. Based on measured changes in absorption spectral slope in relation to solar ultraviolet irradiance, a model was developed which used CDOM as a non-conservative tracer of water masses. Spatial changes in CDOM absorption within the lake were then used to compare photo related transformations to those associated with conservative mixing. PMID- 19303318 TI - Adult neuroectodermal tumors of posterior fossa (medulloblastoma) and of supratentorial sites (stPNET). AB - Medulloblastoma and supratentorial primitive neuroectodermal tumors are rare diseases in adults. Due to this rarity, few prospective clinical trials have been conducted on medulloblastoma in adults, investigations being based exclusively on retrospective studies; the populations considered in literature are small, and the different treatments given span decades, during which diagnostic procedures, neurosurgical skills and radiotherapy techniques have changed. Unlike pediatric patients, adult medulloblastoma patients have been treated according to risk adapted therapeutic strategies in only a few series and despite risk-tailored treatments, 20-30% of patients experience recurrence. Although patients could respond to second line treatments, the prognosis of relapsed patients remains dismal. An important challenge for the future will be the biological characterization of medulloblastoma, with the identification of specific genetic patterns of patients with a better or a worse prognosis. PMID- 19303319 TI - HO-1 and VEGF gene expressions are time dependant during exposure to welding fumes. AB - Hemeoxygenase-1 (HO-1) is a defensive enzyme against oxidative stress. Vascular epithelial growth factor (VEGF) is a potent cytokine which promotes angiogenesis. We used induced sputum (IS) technology to study HO-1 and VEGF expressions in neutrophilic inflammation in asymptomatic welders. Aircraft plant employees were divided into three groups: Welders 1 (n=30) had short-term exposure to aluminum/iron, Welders 2 (n=16) had long-term exposure to cadmium/chromium/iron/nickel, and controls (n=27 non-exposed individuals). Participants underwent pulmonary function tests (PFTs), IS, differential cell counts, and determination of particle size distribution in IS samples. HO-1 and VEGF gene expressions were analyzed by real-time polymerase chain reaction, and protein levels were measured by bilirubin reductase-dependant reaction and ELISA, respectively. All subjects had normal PFTs. Welders 2 had neutrophilic inflammation and higher percentages of particles between 2-5 micron than the other groups. HO-1 inversely correlated with VEGF gene expression: HO-1 was significantly higher and VEGF was significantly lower in the Welders 1 group than in the other groups. There was a correlation between HO-1 expression and protein activity (r=0.33, P=0.05). Particulate matters significantly influenced HO-1 and VEGF gene expressions, caused neutrophilic inflammation and promoted oxidative stress in welders with long-term exposure. PMID- 19303320 TI - Th1 and Th17 balance in inflammatory myopathies: interaction with dendritic cells and possible link with response to high-dose immunoglobulins. AB - To clarify the interactions between dendritic cells (DCs) and Th1 and Th17 T cell subsets and the mode of action of IVIG in inflammatory myopathies, Expression of CD4(+) and CD8(+) T cells, immature (CD1a) and mature (DC-LAMP) DCs, interleukin 17 (IL-17) and interferon-gamma (IFN-gamma), was quantified by immunohistochemistry in muscle biopsies from 13 patients (11 with polymyositis (PM) and 2 dermatomyositis (DM)) obtained before treatment with IVIG. The Th1/Th17 cytokine and the immature/mature DC ratio were studied according to the response to IVIG. Immature DCs were rarely detected compared to mature DCs, observed in all samples except one PM. IFN-gamma-producing cell count was higher than IL-17 count. Neither the expression of IFN-gamma nor IL-17 was correlated with that of DC subsets. Seven of the 13 patients (6 PM and 1 DM) responded to IVIG. T cells and DC subsets were not differentially expressed between responders and non-responders. The frequency of IFN-gamma-producing cells was significantly higher in non-responders with an increased IFN-gamma/IL-17-producing-cell ratio. In conclusion, mature rather than immature DC and IFN-gamma-rather than IL-17 producing cells accumulate in inflamed muscle. Increased IFN-gamma-producing cell count and IFN-gamma/IL-17-ratio were found in IVIG non-responders, suggesting a role for the Th17 mediated pathway in the response to IVIG. PMID- 19303321 TI - Extracellular ATP and P2 receptors are required for IL-8 to induce neutrophil migration. AB - The chemokine interleukin 8 (IL-8) is a major chemoattractant for human neutrophils. Here, we demonstrate novel evidence that IL-8-induced neutrophil chemotaxis requires a concurrent activation of P2 receptors, most likely the P2Y(2) which is dominantly expressed in these cells. Indeed, the migration of human neutrophils towards IL-8 was significantly inhibited by the P2Y receptor antagonists, suramin and reactive blue 2 (RB-2) and potentiated by a P2Y(2) ligand, ATP, but insensitive to specific antagonists of P2Y(1), P2Y(6) and P2Y(11) receptors. Adenosine had no effect on neutrophil migration towards IL-8 which contrasted with the stimulatory effect of this molecule on neutrophil chemotaxis caused by formyl-Met-Leu-Phe (fMLP or fMLF). Taken together, these data suggest that extracellular ATP is necessary for IL-8 to exert its chemotactic effect on neutrophils. PMID- 19303322 TI - Determining the time needed for the vortex method for preparing solvent-free MALDI samples of low molecular mass polymers. AB - Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is an important technique to characterize many different materials, including synthetic polymers. MALDI mass spectral data is used to determine the polymer average molecular weights, repeat units, and end groups. The development of the vortex method of solvent-free sample preparation showed that remarkably short mixing times could prepare samples that yielded high quality MALDI mass spectra. In this paper, we use microscopy images and MALDI mass spectra to evaluate the mixing time required by the vortex method to produce mass spectra for low molecular mass polymer samples. Our results show that mixing times of as little as 10 s can generate homogeneous thin films that produce high quality mass spectra with S/N approximately 100. In addition, ultrashort mixing times of only 2 s still produce samples with mostly smooth morphology and mass spectra with S/N approximately 10. PMID- 19303323 TI - Resistance training alters the sensorimotor control of vasti muscles. AB - The present study examined and compared two modes of weight training (bodybuilding and power-lifting) on the surface EMG of vasti muscles, knee joint position sense and isometric knee extension force in 48 able-bodied subjects. Subjects were randomly allocated into either a moderate loading and repetitions (bodybuilding) training or a high loading and low repetitions (power-lifting) training, or a no training control group. Training was conducted on alternate days with individual supervision. After 8 weeks of training, subjects from both training groups showed significantly earlier EMG onset timing and higher amplitude of vastus medialis obliquus relative to vastus lateralis (p=0.005 or <0.001), and improved knee joint position sense (p<0.001), but no such changes were found in the control group. However, the changes were not significantly different (p>0.05) between the two training groups. The findings suggested that the neuromotor control of the vasti muscles could be altered by regular weight training. PMID- 19303324 TI - Age-related changes of the stretch reflex excitability in human ankle muscles. AB - The purpose of this study was to characterize the effects of aging on the stretch reflex in the ankle muscles, and in particular to compare the effects on the ankle dorsi-flexor (tibialis anterior: TA) and the plantar-flexor (soleus: SOL). Stretch reflex responses were elicited in the TA and SOL at rest and during weak voluntary contractions in 20 elderly and 23 young volunteers. The results indicated that, in the TA muscle, the elderly group had a remarkably larger long latency reflex (LLR), whereas no aging effect was found in the short latency reflex (SLR). These results were very different from those in the SOL muscle, which showed significant aging effects in the SLR and medium latency reflex (MLR), but not in the LLR. Given the fact that the LLR of the TA stretch reflex includes the cortical pathway, it is probable that the effects of aging on the TA stretch reflex involve alterations not only at the spinal level but also at the cortical level. The present results indicate that the stretch reflexes of each of the ankle antagonistic muscles are affected differently by aging, which might have relevance to the neural properties of each muscle. PMID- 19303325 TI - Evaluation and comparison of early hemodynamic changes after elective mitral valve replacement in patients with severe and mild pulmonary arterial hypertension. AB - OBJECTIVE: To evaluate and compare early hemodynamic changes after elective mitral valve replacement (MVR) in patients with severe and mild pulmonary arterial hypertension (PAH). DESIGN: A prospective observational study. SETTING: University-affiliated hospital. PARTICIPANTS: Sixty patients undergoing elective MVR. INTERVENTIONS: The patients were divided into 2 equal groups based on the presence (group A) or absence (group B) of severe PAH defined as systolic pulmonary artery pressure (PAP) > or = 50 mmHg on preinduction pulmonary artery catheterization. Thiopental, fentanyl, midazolam, isoflurane, and rocuronium (or vecuronium if the heart rate >100 beats/min) were used for the induction and maintenance of anesthesia. MVR was performed using standard cardiopulmonary bypass (CPB) techniques. The therapy for PAH was electively instituted in all patients with a nitroglycerin infusion (0.5-1 microg/kg/min), deliberate hypocarbia (arterial carbon dioxide tension < or = 35 mmHg), fractional inspired oxygen concentration = 1.0, and elective ventilation for at least 12 hours in the postoperative period. Hemodynamic and arterial blood gas parameters were serially measured before induction; after intubation; after termination of CPB; after extubation; and at 6, 24, and 48 hours after surgery. Differences in these parameters were analyzed within and among the groups using appropriate statistical tests. MEASUREMENTS AND MAIN RESULTS: The mean CPB and aortic cross clamp times were similar in the 2 groups (78 +/- 33 and 50 +/- 21 minutes in group A and 63 +/- 32 and 41 +/- 23 minutes in group B). The mean PAP, pulmonary capillary wedge pressure, and pulmonary vascular resistance decreased significantly soon after CPB in both groups (p < 0.001), but the decrease was significantly lower in group A (p < 0.001). The mean PAP approached near-normal values in group A (23 +/- 8 mmHg) and normal values in group B (16 +/- 6 mmHg) immediately postoperatively. There was an increase in cardiac index (p < 0.01) after CPB in group A. A relative improvement in oxygenation occurred after MVR in group A compared with group B (p < 0.001). Patients in group A were ventilated for a longer duration (25.9 +/- 18.8 v 17.3 +/- 7.9 hours, p < 0.05). There was no significant difference in the inotropic requirement between the 2 groups. There was no mortality in either group. CONCLUSIONS: PAP returns to near-normal values in patients with severe preoperative PAH and to normal values in patients with mild preoperative PAH immediately after MVR. The outcome after surgery in patients with severe PAH is comparable to those with mild PAH. PMID- 19303326 TI - Transesophageal echocardiography simulator: a new learning tool. PMID- 19303327 TI - Volatile anesthetics reduce mortality in cardiac surgery. AB - OBJECTIVES: A recent meta-analysis suggested that volatile anesthetics reduce postoperative mortality after cardiac surgery. Nonetheless, whether volatile anesthetics improve the outcome of cardiac surgical patients is still a matter of debate. The authors investigated whether the use of volatile anesthetics reduces mortality in cardiac surgery. DESIGN, SETTING, AND INTERVENTIONS: A longitudinal study of 34,310 coronary artery bypass graft interventions performed in Italy estimated the risk-adjusted mortality ratio for each center. A survey was conducted among these centers to investigate whether the use of volatile anesthetics showed a correlation with mortality. MEASUREMENTS AND MAIN RESULTS: All 64 eligible centers provided the required data. The median unadjusted 30-day mortality among participating centers was 2.2% (0.3-8.8), whereas the median risk adjusted 30-day mortality was 1.8% (0.1-7.2). Risk-adjusted analysis showed that the use of volatile anesthetics was associated with a significantly lower rate of risk-adjusted 30-day mortality (beta = -1.172 [-2.259, -0.085], R(2) = 0.070, p = 0.035). Dichotomization into centers using volatile anesthetics in at least 25% of their cases or in less than 25% yielded even more statistically significant results (p = 0.003). Furthermore, a longer use of volatile anesthetics was associated with a significantly lower death rate (p = 0.022); and exploring the impact of the specific volatile anesthetic agent, the use of isoflurane was associated with significant reductions in risk-adjusted mortality rates (p = 0.039). CONCLUSIONS: This survey among 64 Italian centers shows that risk adjusted mortality may be reduced by the use of volatile agents in patients undergoing coronary artery bypass graft surgery. PMID- 19303328 TI - Perioperative beta-blockade in patients undergoing noncardiac surgery: a review of the major randomized clinical trials. PMID- 19303329 TI - Desflurane-induced preconditioning has a threshold that is lowered by repetitive application and is mediated by beta 2-adrenergic receptors. AB - OBJECTIVE: An optimal administration protocol to induce a maximal effect of anesthetic preconditioning has not been evaluated to date. In this study, desflurane preconditioning was characterized with respect to its threshold, dose dependency, and continuous versus repetitive application. Furthermore, the role of beta(2)-adrenergic receptors in anesthetic preconditioning was tested. DESIGN: A randomized controlled study. SETTING: Laboratory study in a University hospital. SUBJECTS: New Zealand white rabbits in vivo. INTERVENTIONS: Systemic hemodynamics were continuously measured. Rabbits were subjected to 30 minutes of coronary artery occlusion and 3 hours of reperfusion. Animals received desflurane continuously for 30 minutes at 0.5, 1.0, or 1.5; desflurane for 90 minutes at 0.5 or 1.5 MAC; or repetitively for three 10-minute periods at 0.5, 1.0, or 1.5 MAC before coronary occlusion. The beta(2)-adrenergic receptor blocker ICI 118,551 (0.2 mg/kg) or saline placebo was given in the absence or presence of 1.0 MAC desflurane. Myocardial infarct size was measured with triphenyltetrazolium staining. MEASUREMENTS AND MAIN RESULTS: Myocardial infarct size was 61% +/- 5% in control experiments. Desflurane, administered continuously at 0.5 MAC for 30 minutes (52% +/- 4%) or 90 minutes (56% +/- 8%) had no effect, whereas 0.5 MAC of desflurane given repetitively reduced infarct size to 36% +/- 7%. Desflurane administered continuously for 30 minutes at 1.0 or 1.5 MAC reduced infarct size to 35% +/- 5% and 39% +/- 4%, respectively. Repetitive application at 1.0 MAC (37% +/- 6%) or 1.5 MAC (29% +/- 4%) and continuous administration of 1.5 MAC for 90 minutes (32% +/- 6%) did not result in further infarct size reduction. ICI 118,551 did not affect infarct size (53% +/- 2%) but abolished desflurane preconditioning (51% +/- 5%). CONCLUSION: beta(2)-Adrenergic receptors mediate desflurane-induced preconditioning. Desflurane-induced preconditioning has a threshold that can be lowered by repetitive administration. PMID- 19303330 TI - Focused cardiovascular ultrasound performed by anesthesiologists in the perioperative period: feasible and alters patient management. AB - OBJECTIVES: The aim of this study was to assess the feasibility and effects on perioperative management of a focused transthoracic echocardiogram performed by anesthesiologists. DESIGN: A prospective observational study of all patients having a focused cardiovascular ultrasound (FoCUS). SETTING: A single tertiary referral university teaching hospital. PARTICIPANTS: Fifty consecutive perioperative patients who had a clinical indication for a FoCUS. INTERVENTIONS: After performing a FoCUS, relevant clinical information was communicated to the anesthesiologist in charge of the case, who then decided on the appropriate management of the patient including the choice of anesthesia, invasive monitoring, fluids, vasoactive drugs, and postoperative care. If indicated, patients were referred for a formal cardiology-based transthoracic echocardiogram. MEASUREMENTS AND MAIN RESULTS: Anesthesiologists were able to obtain diagnostic-quality images during a FoCUS in 98% of patients. The most common indication for a FoCUS was an undifferentiated ejection systolic murmur in 50% of cases, with 38% of all patients having aortic stenosis. In 84% of patients, some change in their perioperative care occurred as a result of the FoCUS study. Major findings correlated with a formal cardiology-based transthoracic echocardiogram in 87% of cases. CONCLUSION: Anesthesiologists with a cardiac and echocardiography background can successfully perform a FoCUS in almost all patients when indicated, which provides valuable new diagnostic information guiding changes in perioperative management in the majority of patients. PMID- 19303331 TI - Erythropoietin response to anemia and its association with autonomic neuropathy in type 2 diabetic patients without advanced renal failure. AB - AIM: We aim to investigate erythropoietin (EPO) response to anemia and its association with autonomic neuropathy in type 2 diabetic patients without advanced renal failure. METHODS: A cross-sectional study was conducted on 211 type 2 diabetes mellitus patients without advanced renal failure [estimated glomerular filtration rate (eGFR) >40 ml/min/1.73 m(2)]. The response of EPO to anemia of type 2 diabetic patients without advanced renal failure was compared with those of nondiabetic control subjects. Autonomic nerve function was assessed using three cardiovascular tests (deep breathing, the Valsalva maneuver, and lying-to-standing). The results of each test were scored as 0 if normal, 1 if borderline, and 2 if abnormal. Autonomic neuropathy was diagnosed when a total score of the tests was 2 or more. RESULTS: Fifty-eight patients were anemic; compared with nonanemic patients, they had a longer duration of diabetes (16.69+/ 10.11 vs. 10.67+/-8.41 years, P<.001), lower eGFR (66.43+/-16.30 vs. 81.74+/ 19.49 ml/min/1.73 m(2), P<.001), and higher cardiovascular autonomic neuropathy score (3.17+/-1.95 vs. 1.79+/-1.72, P<.001). Serum EPO level was weakly correlated with hemoglobin (Hb) level (r=-.085, P<.001). However, the slopes of regression lines between EPO and Hb levels differed significantly between type 2 diabetic patients and nondiabetic control subjects (-0.0085 vs. -0.255, P=.008). Multiple linear regression analysis revealed that cardiovascular autonomic neuropathy score was independently related to Hb (P<.001) or EPO level (P=.052). CONCLUSIONS: Autonomic neuropathy is associated with a blunted EPO response to anemia in type 2 diabetic patients without advanced renal failure. PMID- 19303332 TI - Association of the functional A118G polymorphism of OPRM1 in diabetic patients with foot ulcer pain. AB - BACKGROUND: Diabetic foot ulcer (DFU) patients may experience moderate or severe pain. A single-nucleotide polymorphism, at nucleotide 118 for opioid receptor mu 1 (OPRM1), has been reported to alter the opioid effects to relieve acute or chronic pain. The purpose of this study was to elucidate the correlation between nucleotide 118 variants and foot ulcer pain in DFU patients. METHODS: Sixty-five DFU patients with Grade 2-5 Wagner-Meggitt classification were enrolled. The occurrence of pain in activities was categorized into five grades. Patients were allocated either into the painless DFU group, with a visual analog scale (VAS) pain score or=4 and Grades 3-5 of occurrence of pain in daily activities. DNA was extracted from blood samples of analyzed patients. Using the polymerase chain reaction-single strand conformation polymorphism analysis and DNA sequencing of nucleotide 118, we identified the genotype distribution and allelic frequencies in DFU patients. The sequences of the forward and the reverse primer are designed as follows: 5' TAATACGACTCACTATAGGG-3' and 5'-ACGCACACGATGGAGTAGAG-3', respectively. RESULTS: Fifteen patients were classified into the painful DFU group and 50 patients were classified into the painless DFU group. The amplified DNA fragments showed 26 homozygous (AA), 34 heterozygous (AG), and 5 mutant homozygous (GG) genotypes, with overall A and G allelic frequencies of 66.2% and 33.8%, respectively. The painful DFU group included 10 AA subjects, 4 AG subjects, and 1 GG subject, while the painless DFU group had 16 AA, 30 AG, and 4 GG subjects (P=.038). CONCLUSION: The A118G polymorphism of mu-opioid receptor may be closely associated with DFU pain in 34 out of 50 patients in the painless group and in 5 out of 15 patients in the painful group. This indicates that the nucleotide 118 variant patients may suffer less DFU pain. PMID- 19303333 TI - Prevalence of diabetes, metabolic syndrome, and cardiovascular risk factors in US Asian Indians: results from a national study. AB - BACKGROUND: Although studies of immigrant Asian Indians in other countries show high rates of diabetes (DM), metabolic syndrome (MetS), and cardiovascular disease (CVD), no randomized, population-based studies of this rapidly growing ethnic group exist in the US. METHODS: The sample comprised 1038 randomly selected Asian Indian immigrants, aged 18 years and older at seven US sites. Prevalence of diabetes and MetS (age-adjusted and sex-adjusted means) was estimated and ANOVA was used to calculate gender and group differences (normoglycemia/impaired fasting glucose/diabetes) for CVD risk factors. RESULTS: The mean age was 48.2 years. The majority of respondents were male, married, educated, and with some form of health insurance. Prevalence of diabetes was 17.4%, and 33% of the respondents had prediabetes. Cardiovascular risk factors, especially high levels of triglycerides, total cholesterol, LDL cholesterol, homocysteine, and C-reactive protein, and low levels of HDL cholesterol, were also prevalent; elevated lipoprotein(a) was not observed. The age-adjusted prevalence of MetS was 26.9% by the original NCEP/ATP III criteria, 32.7% by the modified NCEP/ATP III criteria, and 38.2% by the IDF criteria. The MetS rates for women, but not for men, increased with age using all three criteria. There was a progressive worsening of all metabolic parameters as individuals progressed from normal to IFG to diabetes. CONCLUSION: The prevalence rates of diabetes and MetS among US Asian Indians are higher than reported in earlier, nonrandomized, smaller surveys. These data provide a firm basis for future mechanistic and interventional studies. PMID- 19303334 TI - Initial clonal expansion of germinal center B cells takes place at the perimeter of follicles. AB - Current models of the germinal center (GC) response propose that after stimulation at the edges of T cell zones, pre-GC B cells directly migrate to the center of follicles and proliferate to form GCs. We followed the interrelationship of proliferation, differentiation, and microenvironmental locale in populations of pre-GC B cells responding to antigen. In contrast to the predictions of current models, after accumulation at the T-B interface, these cells appeared at the perimeter of follicles adjacent to the marginal zone. There, they rapidly proliferated for several days but underwent no V gene hypermutation and little heavy-chain class switching. Their chemokine receptor expression pattern indicated that these cells were sessile, yet they had begun to acquire many phenotypic characteristics of GC B cells. The expanded clones were subsequently observed in the center of follicles, suggesting that GCs are created by coalescence of B cells from this follicular perimeter response. PMID- 19303335 TI - Observation of molecular migration in porous media using 2D exchange spectroscopy in the inhomogeneous magnetic field. AB - We present a new method for observing fluid diffusion in a porous medium. The method employs 2D exchange spectroscopy for molecules diffusing in the presence of local magnetic field inhomogeneities, in our case distilled water in various sized glass bead packs. Our experiment involves an acquisition and evolution time domain with the two Fourier domains corresponding to the spectral distribution of local fields. We show that exchange in the internal magnetic field can be seen in a 2D spectrum with a characteristic time on the order of that required to diffuse 0.15 sphere diameters with similar behavior found for computer simulations. The method is potentially useful for studying the internal migrations in more complicated systems such as sandstones or other porous media. PMID- 19303336 TI - Stable isotope-enhanced two- and three-dimensional diffusion ordered 13C NMR spectroscopy (SIE-DOSY 13C NMR). AB - The feasibility of obtaining high quality homonuclear or heteronuclear diffusion ordered (13)C NMR data is shown to be greatly improved by using (13)C isotopically-enriched samples. Stable isotope-enhanced diffusion ordered (SIE DOSY) (13)C NMR has been applied to (13)C-enriched carbohydrates, and has been used to determine diffusion coefficients for pentose and hexose monosaccharides, and a disaccharide and trisaccharide. These 2D spectra were obtained with as little as 8 min of acquisition time. Fully resolved 3D DOSY-HMQC NMR spectra of [U-(13)C]xylose, [U-(13)C]glucose, and [1-(13)C(gal)]lactose were obtained in 5h. Sample derivatization with [carbonyl-(13)C]acetate (peracetylation) extends the usefulness of the technique to included non-labeled sugars; the (13)C-carbonyl - carbohydrate ring proton (1)H-(13)C correlations also provide additional structural information, as shown for the 3-D DOSY-HMQC analysis of a mixture of maltotriose and lactose per-[carbonyl-(13)C]acetates. PMID- 19303337 TI - Population pharmacokinetic model for human growth hormone in adult patients in chronic dialysis compared with healthy subjects. AB - OBJECTIVE: To develop a population pharmacokinetic (PK) model of recombinant human growth hormone (rhGH) treatment in patients with end-stage renal disease (ESRD) and healthy volunteers (HVs), to support future study design. DESIGN: This was an open, non-randomized, single-centre parallel-group study lasting 8-9 days. Various compartment models with first-order and Michaelis-Menten absorption and elimination were explored. Eleven adult ESRD patients and 10 matched HVs received 50 microg/kg/day rhGH (subcutaneous (s.c.) injection) for 8 or 7 days, respectively. Blood samples were drawn every 30 min for 24h following dosing on Days 0, 7 and 8 (ESRD patients). Influence of the covariates subject group (ESRD/HV), gender, weight, and dialysis flow-rate on model parameters was examined. RESULTS: The final model was one-compartmental with Michaelis-Menten absorption and elimination. The following estimates were obtained: maximum absorption rate (VMA) - 11.3 microg/kg/h (both groups); amount of drug corresponding to half-maximum absorption rate (KMA) - 1.06 and 18.8 microg/kg (ESRD patients and HVs, respectively; P<0.001); maximum elimination rate (VM) - 9.37 and 13.0 microg/kg/h (ESRD patients and HVs, respectively; P<0.001); amount of drug corresponding to half-maximum elimination rate - 18.9 microg/kg (both groups). Significant differences in KMA and VM between HVs and ESRD patients corresponded to higher absorption and lower elimination rates in ESRD, but all GH profiles were back to baseline by 20-22h and no overall accumulation occurred. Simplified posterior predictive checks indicated that the model satisfactorily captured PK. All non-compartmental estimates for AUC(0-24h) and C(max) lay within 95% confidence limits of the simulated distributions. CONCLUSIONS: A population PK model was established, which showed acceptable performance for trial simulation purposes. PMID- 19303338 TI - How do patients with type 2 diabetes perceive their disease? Insights from the French DIABASIS survey. AB - AIM: The main purpose of this survey was to describe type 2 diabetes (T2DM) from the patient's standpoint in a representative French panel in 2008. METHODS: Fourteen thousand two hundred and one individuals from the general population aged 45 or older completed a self-questionnaire exploring knowledge about diabetes; 1092 replies were from patients with T2DM. RESULTS: The prevalence of T2DM in this population was 7.7%, with demographics as follows: 60% men; mean age: 66 years; mean age at diagnosis: 55 years; mean BMI: 29 kg/m(2). Eighty-five percent of T2DM patients reported that they wanted more information about at least one aspect of the disease at diagnosis; they reported feeling anxious (30%), frightened (13%), angry (4%) or that the disease was unfair (12%). Half of the patients had modified their dietary habits but 71% found it difficult to engage in regular physical activity. Most patients (90%) were treated with drugs: 81% with oral antidiabetic drugs (OAD) (44% in monotherapy) while 19% received insulin (alone or in combination with OAD). Twenty-three percent complained of weight gain since start of current therapy (average gain of 7.3 kg). Insulin initiation represented a turning point for patients who became more aware of the disease severity, more willing to follow advice and to take greater control over their disease management. The mean time from diagnosis to insulin initiation was 13.8 years. Half of the patients perceived their disease as severe especially women, patients who initially reacted with anxiety, insulin-treated patients and those actively involved in their disease management. Some gender differences emerged: women took the disease more seriously, were more engaged in self management, and reported a higher impact on daily life. CONCLUSIONS: DIABASIS provides important information for diabetes care by highlighting patients' views of the disease, such as distress at diagnosis, lack of adequate information to cope with this distress and the important supportive role played by the family. A deeper understanding of patients' perception of the disease would help optimize customized care. PMID- 19303339 TI - [Impact of radiotherapy modalities on local control and survival in adjuvant breast cancer treatment]. AB - Conventional radiotherapy (RT) in breast cancer treatment includes a total dose of 50Gy with a 16Gy-boost in a smaller volume. After mastectomy, radiotherapy of all node-positive patients seems to be applied whatever the positive node number. Modulation of RT technique uses the fraction dose, namely through accelerated hypofractionation regimen in a constant volume. A group of patient with a low risk of recurrence may benefit from this technique. In a smaller irradiated volume, accelerated hypofractionation still belongs to intensive clinical research evaluating intraoperative RT, brachytherapy, Mammosite, and external partial breast irradiation. Intensity-modulated radiotherapy decreases the risk of moist desquamation. The development of the gating offers greater protection of certain critical organs including lung, heart, and chest/ribs. Finally, predictive tests identifying patients at high risk of developing late effects led us to adapt RT technique to some of them and therefore increase long term therapeutic ratio. PMID- 19303340 TI - Unexpected remission of renal AA amyloidosis associated with rheumatoid arthritis. PMID- 19303341 TI - Marked hypertriglyceridemia upon treatment with etanercept. PMID- 19303342 TI - Late Mycobacterium bovis spondylitis after intravesical BCG therapy. AB - A 72-year-old man presented with a 6-month history of low back and leg pain. Past medical history revealed transurethral resection of bladder cancer followed by multiple intravesical BCG instillation 12 years ago. Imaging studies of the thoracolumbar spine showed osteolysis of the L3 and L4 vertebrae and the associated intervertebral disc space, and a large soft tissue mass with signal abnormalities suggesting of an abscess. CT-guided needle biopsy showed Mycobacterium bovis infection. A triple anti-tuberculous chemotherapy regimen including isoniazid, rifampicin, and ethambutol was administered for 12 months. Surgical treatment included drainage of the abscess and L2-L5 spinal instrumentation and fusion. Intravesical BCG therapy may be complicated by late disseminated disease to the bone even many years after initial BCG therapy. Patients having BCG therapy should be closely evaluated thereafter for the possibility of hematogenous spread of mycobacteria to distant sites. PMID- 19303343 TI - Compared imaging of the rheumatoid cervical spine: prevalence study and associated factors. AB - INTRODUCTION: Cervical spine involvement is common and potentially severe in patients with rheumatoid arthritis (RA). The objectives of this study were to compare the prevalences of cervical spine abnormalities detected by standard radiography, computed tomography (CT), and magnetic resonance imaging (MRI) in patients with RA; and to identify factors associated with cervical spine involvement. METHODS: We studied 40 patients who met American College of Rheumatology criteria for RA and had disease durations of 2 years or more. Each patient underwent a physical examination, laboratory tests, standard radiographs (anteroposterior, lateral, open-mouth, flexion, and extension views), MRI with dynamic maneuvers in (if not contraindicated), and CT. RESULTS: Cervical spine involvement was found by at least one imaging technique in 29 (72.5%) patients (standard radiography, 47.5%; CT, 28.2%; and MRI, 70%) and was asymptomatic in 5 (17.2%) patients. C1-C2 pannus was the most common lesion (62.5% of cases), followed by atlantoaxial subluxation (AAS, 45%). The most common AAS pattern was anterior subluxation (25%), followed by lateral subluxation (15%) then by vertical, rotatory, and subaxial subluxations (10% each). Erosions of the dens were seen in 67.5% of patients by MRI, 41% by CT, and 12.5% by standard radiography. Of the 10 cases of anterior AAS by any modality, 9 were detected by standard radiography and 7 by MRI. CT was the best technique for visualizing atypical rotatory or lateral AAS. MRI was best for assessing the C1-C2 pannus, dens erosions, and neurologic impact of the rheumatoid lesions. The comparison of patients with and without cervical spine lesions suggested that higher modified Sharp score and C-reactive protein values predicted cervical spine involvement (P=0.002 and P=0.004, respectively). CONCLUSION: Cervical spine involvement is common and may be asymptomatic, indicating that routine cervical spine imaging is indicated in patients with RA. Standard radiography including dynamic views constitutes the first-line imaging method of choice. Sensitivity and comprehensiveness of the assessment are greatest with MRI. MRI and CT are often reserved for selected patients. Cervical spine involvement is associated with disease activity and with rapidly progressive joint destruction. PMID- 19303344 TI - Chin-sternum-heart syndrome type of injury observed in a pedestrian victim of car traffic accident. AB - We report an autopsy case of a pedestrian victim of car traffic accident with the chin-sternum-heart syndrome type of injury. A drunken man who lay on the road was run over by a car. He died immediately at the scene. The autopsy findings were as follows: large scalp lacerations, abrasions in the chin and the sternal region, a transverse fracture of the sternum, ruptures of the heart, ruptures of the ascending and descending aortae, rupture and hemorrhage of the nuchal muscle, ring fracture of the base of the skull, subarachnoid hemorrhage at the base of the brain, multiple rib fractures, anterior compression fracture of the 11th thoracic vertebra, and small lacerations of the liver. Blood ethanol level was 2.92 mg/g. These findings indicate that there was hyperflexion of the neck and then the victim's heart was strongly compressed and ruptured by the collision of the chin with the sternal region of the chest. PMID- 19303345 TI - How rapidly does cerebral swelling follow trauma? Observations using an animal model and possible implications in infancy. AB - To study the speed of development of cerebral edema in an animal model, five 2 year-old male anesthetized Merino sheep were impacted in the left temporal region by a humane stunner. Following the induction of blunt craniocerebral trauma a highly significant increase in intracranial pressure (ICP) values occurred relative to control animals (ANOVA, p<0.001). An immediate increase in ICP to values over 20 mmHg occurred within the first 30 min, and by 60 min, mean ICP was over 25 mmHg (p<0.05 versus controls). ICP continued to increase with time such that by 4h after injury, values were consistently greater than 30 mmHg (p<0.001 versus controls). The mean brain tissue oxygenation (P(bt)O(2)) in control animals over the 4h monitoring period was 52+/-8 mmHg with a highly significant (ANOVA, p<0.001) and early decrease in P(bt)O(2) recorded in impacted animals following craniocerebral trauma. By 30 min after injury, P(bt)O(2) was approximately 35 mmHg, while by 60 min, it had decreased to 20+/-2 mmHg (p<0.001 versus controls). The P(bt)O(2) continued to decline with time such that by 4h, the value was 14+/-2 mmHg (p<0.01 versus controls), or 27% of the mean control values. These data demonstrate that vasoreactive changes with increased blood volume and interstitial transfer of fluid may occur rapidly within brain tissue after blunt trauma. Detection of cerebral swelling at autopsy may not, therefore, imply prolonged survival. PMID- 19303346 TI - Forensic implication of muscle level of nicotine in an adipocere body found in the sea. AB - We reported previously that blood levels of nicotine in suicidal smokers tend to be significantly higher than those in non-suicidal smokers, and blood level of nicotine seems to be a useful criterion for discriminating suicide from other types of death. In this paper, we report nicotine and cotinine levels in various tissues of an adipocere body found in the sea. The cause of death was drowning, and the postmortem time interval was approximately 5 months at autopsy. His driver's license was concealed in his sock, which seemed to suggest that he committed to suicide. In toxicological analysis by gas chromatography, nicotine and cotinine in the femoral muscle were detected at concentrations of 213 and 488 ng/g, respectively, and these substances were also detected in the brain, liver and kidney. For evaluating the tissue levels of nicotine and cotinine in the adipocere body, we analyzed these levels in blood and various tissues of 13 autopsy cases of smokers. Nicotine and cotinine levels in blood were the most similar to those in skeletal muscle. Although the postmortem time interval, the formation process of adipocere and the environmental condition in water may affect nicotine and cotinine levels in the femoral muscle, the high muscle level of nicotine in the present case seem to implicate suicidal death. PMID- 19303347 TI - 128-channel somatosensory evoked potentials in the differential diagnosis of parkinsonian disorders. AB - PURPOSE: The differential diagnosis of parkinsonian disorders can be very difficult, especially at an early stage. In this study, we investigated whether SEP amplitude recorded by 128-channel EEG is useful for diagnosis of parkinsonian disorders, and in particular whether SEP asymmetry can differentiate corticobasal degeneration (CBGD) from other parkinsonian disorders. METHODS: We recorded median nerve SEPs in 47 patients suspected of CBGD, supranuclear palsy or definite Parkinson's disease at an early stage. We compared SEP asymmetry and parietal peak amplitudes of the patients after grouping them based on their clinical diagnosis after 1-5 years of follow-up. In nine subjects the diagnosis remained unclear. RESULTS: Three of 13 patients with a clinical diagnosis of CBGD had an abnormal SEP asymmetry. Furthermore, we found extremely high N20 amplitudes in three other patients with CBGD. However, similar asymmetry abnormalities were found in patients with other Parkinsonian disorders. CONCLUSION: Despite the use of 128-channel SEP recordings and analysis techniques, which are more accurate than conventional techniques, sensitivity and specificity of cortical median nerve SEP asymmetry and parietal amplitude for differentiating CBGD from other parkinsonian disorders were low at an early stage of the disease. A possible reason for this may be that the hand area of the primary somatosensory cortex was not yet affected in most CBGD patients. PMID- 19303348 TI - Singlet oxygen in plants: production, detoxification and signaling. AB - Singlet oxygen ((1)O(2)) is a singular reactive oxygen species (ROS) that is produced constitutively in plant leaves in light via chlorophylls that act as photosensitizers. This (1)O(2) production is spatially resolved within thylakoid membranes and is enhanced under light stress conditions. (1)O(2) can also be produced by phytotoxins during plant-pathogen interactions. (1)O(2) is highly reactive, can be toxic to cells and can be involved in the signaling of programmed cell death or acclimation processes. Here, we summarize current knowledge on (1)O(2) management in plants and on the biological effects of this peculiar ROS. Compared with other ROS, (1)O(2) has received relatively little attention, but recent developments indicate that it has a crucial role in the responses of plants to light. PMID- 19303349 TI - Janus-faced charge recombinations in photosystem II photoinhibition. AB - Light-induced damage of the photosynthetic apparatus in plants is an important phenomenon that primarily affects the photosystem II complex. Here, we propose a new model of photoinhibition in which charge recombination processes have a double-faced role: first, photodamage is induced by singlet oxygen, which is produced via interaction with the triplet reaction center chlorophyll ((3)P(680)) arising from the recombination of the charge-separated state between P(680) and the pheophytin electron acceptor ((3)[P(680)(+)Phe(-)]). Second, photoprotection is provided by competition between (3)[P(680)(+)Phe(-)] formation and direct recombination of the (1)[P(680)(+)Phe(-)] and P(680)(+)Q(A)(-) states. The efficiency of these two pathways is under control of the redox potential of the Phe and Q(A) electron acceptors, which is utilized during adaptation to high light conditions. PMID- 19303350 TI - The many functions of ERECTA. AB - The Arabidopsis thaliana accession Landsberg erecta contains an induced mutation in the leucine-rich repeat receptor-like Ser/Thr kinase gene ERECTA. Landsberg erecta is commonly used as a genetic background in mutant screens and in natural variation studies. Therefore, the erecta mutation is present in many loss-of function mutants and recombinant inbred lines. Information on how the absence of functional ERECTA affects the interpretation of obtained phenotypic results is scattered. In this report we inventoried ERECTA functions and highlight ERECTA as a pleiotropic regulator of developmental and physiological processes, as well as a modulator of responses to environmental stimuli. PMID- 19303351 TI - Congenital aortic disease: 4D magnetic resonance segmentation and quantitative analysis. AB - Automated and accurate segmentation of the aorta in 4D (3D+time) cardiovascular magnetic resonance (MR) image data is important for early detection of congenital aortic disease leading to aortic aneurysms and dissections. A computer-aided diagnosis (CAD) method is reported that allows one to objectively identify subjects with connective tissue disorders from 16-phase 4D aortic MR images. Starting with a step of multi-view image registration, our automated segmentation method combines level-set and optimal surface segmentation algorithms in a single optimization process so that the final aortic surfaces in all 16 cardiac phases are determined. The resulting aortic lumen surface is registered with an aortic model followed by calculation of modal indices of aortic shape and motion. The modal indices reflect the differences of any individual aortic shape and motion from an average aortic behavior. A Support Vector Machine (SVM) classifier is used for the discrimination between normal and connective tissue disorder subjects. 4D MR image data sets acquired from 104 normal volunteers and connective tissue disorder patients MR datasets were used for development and performance evaluation of our method. The automated 4D segmentation resulted in accurate aortic surfaces in all 16 cardiac phases, covering the aorta from the aortic annulus to the diaphragm, yielding subvoxel accuracy with signed surface positioning errors of -0.07+/-1.16 voxel (-0.10+/-2.05mm). The computer-aided diagnosis method distinguished between normal and connective tissue disorder subjects with a classification correctness of 90.4%. PMID- 19303352 TI - Synthesis and spectroscopic studies of some cadmium(II) and mercury(II) complexes of an asymmetrical bidentate Schiff base ligand. AB - Synthesis and spectroscopic studies on four-coordinate complexes of cadmium(II) and mercury(II) halides with a new asymmetrical bidentate Schiff base ligand of N,N'-bis[alpha-methylcinamaldehydene]propane-1,2-diamine(L) are described. The ligand and its complexes were characterized by elemental analysis, molar conductance, UV-visible spectra, FT-IR spectra, MS, (1)H NMR and (13)C NMR spectra. The complexes are non-electrolytes in DMF. The electronic spectra of the complexes were recorded in DMF solution. (1)H and (13) C NMR spectra been studied in CDCl(3). The molar conductance as well as spectral properties indicated the complexes do not dissociate in DMF and retain their coordination. FT-IR and NMR spectra of the complexes exhibit downfield as well as upfield shifts of the free ligand resonances that show change in geometry during the coordination. The suggested structure of the complexes is pseudo-tetrahedral. Molecular structures of the complexes have been optimized by MM+ calculations that supported pseudo tetrahedral geometry around the metal (II) ions. PMID- 19303353 TI - FTIR and Raman spectra, DFT and normal coordinate computations of 2,4,5- and 2,4,6-tri-fluoroanilines. AB - FTIR and Raman spectra of the 2,4,5- and 2,4,6-tri-fluoroaniline molecules have been reported. Density functional method has been employed to calculate the optimized geometrical parameters, atomic charges and vibrational frequencies. In addition, SQM force field method has also been employed to calculate potential energy distribution matrix. The observed and calculated IR and Raman spectra have been simulated. Each normal mode has been assigned using observed and calculated vibrational frequencies, IR and Raman intensities, depolarization ratios for the Raman lines, vector displacements and potential energy distributions. Optimized geometrical parameters, atomic charges and vibrational frequencies have been compared for the title molecules. The influences of presence of fluorine atoms to the geometries and normal modes of the aniline molecule have also been discussed. PMID- 19303354 TI - Spectroscopic, structural and theoretical studies of novel, potentially cytotoxic 4-acridonecarboxamide imines. AB - Ten novel, potentially intercalating 4-acridonecarboxamide azomethines and ketimines have been prepared by the condensation reaction of 9-oxo-9,10 dihydroacridine-4-carboxylic acid hydrazide with various aldehydes and ketones. The structures of the compounds were characterized spectroscopically by NMR ((1)H, (13)C, (15)N nuclei and 2D experiments), UV-vis, IR and fluorescence methods and by quantum chemical calculations using DFT at the B3LYP/6-311+G(d,p) level of theory and semiempirical ZINDO and AM1 methods. NMR chemical shift variations for C-4' were assessed due to changes in the polarizability of the imine C(4')=N(3') bond rather than direct inductive effects arising from the C-4' substituents. In concert with this was the reversed order observed for the N-3' chemical shifts with DFT-calculated atomic charges confirming the bond polarization. Both intra- and intermolecular hydrogen bonds between the acridone NH hydrogen and the amidic carbonyl oxygen were found to exist by FT-IR spectroscopy. Quantum chemical calculations were used to evaluate the configurational, tautomeric, conformational and hydrogen bonding states of the molecules as well as predict the NMR and IR data. The hypsochromic shifts observed in the UV-vis spectra upon changing from m-cresol to DMA, DMF or methanol were evaluated in terms of solvent polarity (giving rise to solvated excited state destabilization) and solvent aromaticity (giving rise to solvated excited state stabilization). The fluorescence of the compounds were modest, except for the 2,6-dichloro derivative, with respect to 9-isothiocyanatoacridine. PMID- 19303355 TI - Anticipatory postural control associated with bilateral arm flexion and event related potential in a Kanji Stroop-like task. AB - OBJECTIVE: We investigated anticipatory postural control during arm flexion and event-related potential (ERP) in a Kanji Stroop-like task. METHODS: The Kanji for RED or BLUE colored in red, blue, or black was randomly presented on a screen for 100 ms. Subjects flexed their arms in response to target stimuli colored black (control condition) or which were congruent or incongruent with the meaning of Kanji characters. P300 of ERPs elicited by the target stimuli and onset timing of electromyographic activity in arm and postural muscles were measured. RESULTS: Reaction time of the anterior deltoid (AD) was shorter and P300 amplitude larger in the order of control, congruent, and incongruent conditions. A distinct P515 component was recognized in ERPs under congruent and incongruent conditions. No significant difference among conditions was observed in onset timing of erector spinae, biceps femoris, or gastrocnemius activity with respect to AD. CONCLUSIONS: Changes in cognitive processing and allocation of attentional resources according to task demands influence the onset timing of arm movement, but not that of anticipatory postural control, suggesting the presence of fixed synergy of arm movement and postural control. SIGNIFICANCE: These findings are important for understanding the effects of cognitive function on anticipatory postural control. PMID- 19303356 TI - Traditional needle acupuncture treatment for insomnia: a systematic review of randomized controlled trials. AB - OBJECTIVES: Previous reviews regarding traditional needle acupuncture (TNA) treatment for insomnia were limited to English scientific literature. A comprehensive review including Chinese and English literature has therefore been conducted to examine the efficacy of TNA for insomnia. METHODS: We performed systematic review of randomized controlled trials (RCTs) of TNA as intervention for insomnia against placebo, Western medication, and non-treated controls. The methodological quality of the studies was assessed by the modified Jadad score and the acupuncture procedure was appraised by the STRICTA criteria. RESULTS: Twenty RCTs were identified for detailed analysis. Majority of the RCTs concluded that TNA was significantly more effective than benzodiazepines for treating insomnia, with mean effective rates for acupuncture and benzodiazepines being 91% and 75%, respectively. In two more appropriately conducted trials, TNA appeared to be more efficacious in improving sleep than sleep hygiene counseling and sham acupuncture. Standardized and individualized acupuncture had similar effective rates. Despite these positive outcomes, there were methodological shortcomings in the studies reviewed, including imprecise diagnostic procedure, problems with randomization, blinding issues, and insufficient safety data. Hence, the superior efficacy of TNA over other treatments could not be ascertained. CONCLUSION: Since the majority of evidence regarding TNA for insomnia is based on studies with poor quality research designs, the data, while somewhat promising, do not allow a clear conclusion on the benefits of TNA for insomnia. Moreover, the results support the need for large scale placebo-controlled double-blinded trials. PMID- 19303357 TI - Disturbed sleep and musculoskeletal pain in the bed partner of patients with obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) has been recognized as a distressing experience to the female partner, but a causal association remains controversial. Furthermore, a growing body of evidence supports a relationship of distress and sleep disruptions with musculoskeletal pain, also prevalent in middle-aged women. To test the hypothesis that sharing a bed with an OSA man may contribute to manifestations of distress and impaired sleep, we conducted a case-control study of 17 OSA wives and 17 wives of healthy sleepers. METHODS: Clinical outcome variables were scores of the Fibromyalgia Impact Questionnaire (FIQ) and visual analog scales (VAS), tender point (TP) count and algometric index. Sleep outcome parameters were VAS scores for sleep quality and polysomnographic (PSG) parameters. Secondary outcomes were VAS scores for distress and marital relationship and coping strategies adopted by OSA wives. Clinical assessment and PSG scoring were conducted blindly. RESULTS: After controlling for age and menopausal status, OSA wives exhibited lower sleep quality and higher distress scores than controls (p<0.05, all). Increase in pain threshold (TP count and algometric index) and in FIQ score were also observed, and in their PSG, there was an increase in awaken period and stage 1 amount during sleep, as well as in alpha power during slow wave sleep (p<0.05, all). These sleep parameters had substantial correlation with tiredness and poor sleep quality and were moderately correlated to pain assessments and distress scores. CONCLUSION: Thus, independently of age and menopausal status of the group, wives of OSA patients exhibited an increase in pain threshold, distress and impaired sleep in comparison to controls. PMID- 19303358 TI - Balancing AID and DNA repair during somatic hypermutation. AB - Somatic hypermutation (SHM) of Ig genes in B cells is crucial for antibody affinity maturation. The reaction is initiated by cytosine deamination of Ig loci by activation induced deaminase (AID) and is completed by error-prone DNA repair enzyme processing of AID-generated uracils. The mechanisms that target SHM specifically to Ig loci are poorly understood. Recently, it has been demonstrated that although AID preferentially targets Ig loci, it acts surprisingly widely on non-Ig loci, many of which are protected from mutation accumulation by high fidelity DNA repair. We propose that breakdown of this high fidelity repair process helps explain oncogene mutations observed in B-cell tumors, and further, that many oncogenes are vulnerable to AID-mediated DNA breaks and translocations in normal activated B cells. PMID- 19303359 TI - Endothelial progenitor cell-based neovascularization: implications for therapy. AB - Ischemic cardiovascular events are a major cause of death globally. Endothelial progenitor cell (EPC)-based approaches can result in improvement of vascular perfusion and might offer clinical benefit. However, although functional improvement is observed, the lack of long-term engraftment of EPCs into neovessels has raised controversy regarding their mechanism of action. We and others have hypothesized that after ischemic injury, EPCs induce neovascularization through the secretion of cytokines and growth factors, which act in a paracrine fashion and induce sprouting angiogenesis by the surrounding endothelium. In this concise review, we discuss the (patho)physiology of EPC induced neovascularization and focus on the paracrine signals secreted by EPCs and the effects they elicit. In future therapies, clinical administration of these paracrine modulators using slow-release depots might induce neovascularization and might therefore hold promise for vascular regenerative medicine. PMID- 19303360 TI - Mitogen-activated protein kinases in male reproductive function. AB - Recent studies have shown that male reproductive function is modulated via the mitogen-activated protein kinase (MAPK) cascade. The MAPK cascade is involved in numerous male reproductive processes, including spermatogenesis, sperm maturation and activation, capacitation and acrosome reaction, before fertilization of the oocyte. In this review, we discuss the latest findings in this rapidly developing field regarding the role of MAPK in male reproduction in animal models and in human spermatozoa in vitro. This research will facilitate the design of future studies in humans, although much work is needed before this information can be used to manage male infertility and environmental toxicant-induced testicular injury in men, such as blood-testis-barrier disruption. PMID- 19303361 TI - Bronchopulmonary dysplasia and emphysema: in search of common therapeutic targets. AB - Bronchopulmonary dysplasia of the premature neonate and emphysema of the adult lung are common diseases that are characterized by increased airspace size and respiratory insufficiency and that presently lack efficient treatment. Although the former leads to impaired alveolar development and the latter to alveolar destruction, they have striking similarities in their pathophysiology, including the precipitating effect of oxidative stress, sustained inflammation, enhanced apoptosis, protease-antiprotease imbalance, elastic fiber deterioration and altered microvascularization. This review aims to comparatively analyze their molecular mechanisms to try identify common therapeutic targets. The recent discovery that alveolar developmental and maintenance programs share the same signal molecules and pathways, together with considerable increase in their understanding, have facilitated the development of common innovative strategies that have started to be tested in experimental models and pilot clinical studies. PMID- 19303362 TI - Degenerative diseases, oxidative stress and cytochrome c oxidase function. AB - Aging and degenerative diseases are associated with increased levels of reactive oxygen species (ROS). ROS are mostly produced in mitochondria, and their levels increase with higher mitochondrial membrane potential. Cellular respiratory control is based on inhibition of respiration by high membrane potentials. However, we have described a second mechanism of respiratory control based on allosteric inhibition of cytochrome c oxidase (CcO), the terminal enzyme of the respiratory chain, at high ATP:ADP ratios. The mechanism is independent of membrane potential. We have proposed that feedback inhibition of CcO by ATP keeps the membrane potential and ROS production at low levels. Various forms of stress switch off allosteric ATP-inhibition via reversible dephosphorylation of CcO, resulting in increased membrane potential and cellular ROS levels. This mechanism is proposed to represent a missing molecular link between stress and degenerative diseases. PMID- 19303363 TI - The coronary artery bypass graft surgery trajectory: Gender differences revisited. AB - Over the past several decades there has been substantial research interest in gender differences within the coronary artery bypass graft (CABG) surgery trajectory. However, the debate persists regarding the reasons why women may have less favorable outcomes. As part of a larger study, we explored gender differences in the physiological and psychosocial dimensions of pre-operative status, and post-operative morbidity and quality of life outcomes in CABG surgery patients. A purposive sample of patients on the waiting list for CABG surgery (N=195; 157 males; 38 females) was followed for 6 months post-surgery. The results reflected consistent evidence of a male advantage across the CABG surgery trajectory. Though gender differences in age were non-significant, females had significantly more post-operative respiratory complications (p=0.005), a longer hospital stay (p=0.003), more symptoms at 2 weeks post-discharge, and a lower quality of life at 6 weeks and 6 months post-discharge. Our findings provide important insights for improving CABG surgery outcomes for both men and women. In particular, implementing creative strategies to improve physical functioning pre operatively, may improve post-operative quality of life outcomes in this population. PMID- 19303364 TI - Improving uptake of cardiac rehabilitation: using theoretical modelling to design an intervention. AB - BACKGROUND: Attendance rates at cardiac rehabilitation remain low with typically fewer than 35% of eligible patients attending. Much of the poor attendance stems from invited patients failing to attend. AIM: To design a theoretically based intervention to improve attendance at cardiac rehabilitation. METHODS: Our methods followed recommendations that have been developed from the Medical Research Council (MRC) framework for the design of complex interventions. We conducted three processes that progressed simultaneously: 1) literature review for evidence on epidemiology, behavioural theory, and efficacy of interventions; 2) expert meetings on behavioural theory and to select target points for intervention; and 3) development and theoretical modelling of the intervention. RESULT: Our final interventions were a theoretically worded invitation letter and leaflet based on the Theory of Planned Behaviour and the Common Sense Model of Illness, designed to: a) motivate patients through professional recommendation; b) provide simple information on the contents of cardiac rehabilitation emphasising ease for participants; c) reassure participants that the programme is tailored to their personal needs in a safe supervised environment; and d) reinforce the benefits of attending cardiac rehabilitation. CONCLUSION: A theoretically worded letter and leaflet could be an inexpensive intervention to improve attendance at cardiac rehabilitation. The letters and leaflets will now be evaluated in a randomised trial. PMID- 19303365 TI - Pediatric hemorrhagic cystitis. AB - PURPOSE: To review the current literature as it pertains to hemorrhagic cystitis (HC) in the pediatric bone-marrow transplant (BMT) population. By reviewing the pathophysiology of the disease, preventive methods, and therapeutic options, urologists may be better equipped to manage this challenging clinical scenario. MATERIALS AND METHODS: The HC literature was reviewed using a MEDLINE/PubMed literature search, specifically focusing on the pediatric BMT population as it pertains to the incidence, pathophysiology, prevention, and treatment of HC. RESULTS: Conservative estimates of HC incidence in recent retrospective studies of pediatric BMT populations still approach 10-20%. Several high-volume pediatric BMT centers have reported contemporary data on their experience with HC providing increased insight into incidence and pathophysiology. Accumulating evidence linking BK virus to HC is a significant development warranting further investigation. Other contributing agents/risk factors need identification in the likely multifactorial etiology of HC. Preventive and therapeutic strategies have made modest advances, but certainly need further validation with prospective randomized studies. CONCLUSIONS: Pediatric BMT patients are susceptible for HC development despite preventive measures and improved insight into the pathophysiology. Unfortunately, there are no evidence-based treatment guidelines for this difficult clinical issue that frequently requires prolonged care and multiple treatment modalities necessitating judicious patience in the application of more aggressive interventions. PMID- 19303366 TI - Dynamics of T cell activation accompanying CD4 recovery in antiretroviral treated HIV-infected Ugandan children. AB - Africans have elevated T cell activation compared to residents of Europe or the USA. Levels of T cell activation also correlate with disease progression in HIV infected individuals. We sought to determine if treatment with antiretroviral therapy (ART) would reduce levels of T cell activation (CD38 and HLADR co expression) in HIV-infected Ugandan children. The median CD8+ T cell activation level among 199 ART-treated children (30%) was lower than in 57 ART-naive children (45%, p<0.001), but remained higher than in 30 HIV-uninfected children (18%, p<0.001). Among ART-treated children, CD4% was inversely correlated with both CD8- (rho=-0.61, p<0.001) and CD8+ (rho=-0.38, p<0.001) T cell activation. Prospectively, CD4 recovery correlated with post-treatment CD8+ T cell activation level (p=0.008). Our data suggest that significant decreases in T cell activation accompany CD4 recovery in ART-treated HIV-infected African children, to levels that approach but do not reach those of uninfected children. PMID- 19303367 TI - Characterising pelvic masses using ultrasound. AB - The evaluation of a pelvic mass, detected at routine ultrasound or in the context of having acute symptoms, requires certain methodical steps. It is necessary to diagnose the primary site or origin of the lesion; understand whether the lesion is an intra- or an extra-peritoneal lesion; distinguish whether the structure of interest is 'functional' and transitory or if we are dealing with a persistent mass; attempt to discriminate between the benign or malignant nature of the lesion; formulate a specific diagnosis; and furthermore, in the case of a possible malignant mass, define the stage of the disease process. Transvaginal ultrasonography is a dynamic and interactive examination and besides an analysis of the echostructure, and 'elasticity' of a pelvic mass, it also permits an assessment of site-specific pain in different pelvic areas, and an evaluation of the movement of the mass in relation to adjacent structures. All these 'dynamic' features, together with morphological and vascular parameters, are essential for making a correct diagnosis. The features of acute ovarian pathology including torsion are not discussed in detail here. This article will concentrate on the characterisation of pelvic masses that may be encountered by any examiner in the course of an assessment of the female pelvis. PMID- 19303368 TI - Maternal near miss--towards a standard tool for monitoring quality of maternal health care. AB - Maternal mortality is still among the worst performing health indicators in resource-poor settings. For deaths occurring in health facilities, it is crucial to understand the processes of obstetric care in order to address any identified weakness or failure within the system and take corrective action. However, although a significant public health problem, maternal deaths are rare in absolute numbers especially within an individual facility. Studying cases of women who nearly died but survived a complication during pregnancy, childbirth or postpartum (maternal near miss or severe acute maternal morbidity) are increasingly recognized as useful means to examine quality of obstetric care. Nevertheless, routine implementation and wider application of this concept in reviewing clinical care has been limited due to the lack of a standard definition and uniform case-identification criteria. WHO has initiated a process in agreeing on a definition and developing a uniform set of identification criteria for maternal near miss cases aiming to facilitate the reviews of these cases for monitoring and improving quality of obstetric care. A list of identification criteria was proposed together with one single definition. This article presents the proposed definition and the identification criteria of maternal near miss cases. It also suggests procedures to make maternal near miss audits operational in monitoring/evaluating quality of obstetric care. The practical implementation of maternal near miss concept should provide an important contribution to improving quality of obstetric care to reduce maternal deaths and improve maternal health. PMID- 19303369 TI - Early termination of spiral wave reentry by combined blockade of Na+ and L-type Ca2+ currents in a perfused two-dimensional epicardial layer of rabbit ventricular myocardium. AB - BACKGROUND: Modification of spiral wave (SW) reentry by antiarrhythmic drugs is a central issue to be challenged for better understanding of their benefits and risks. OBJECTIVE: We investigated the effects of pilsicainide and/or verapamil, which block sodium and L-type calcium currents (I(Na) and I(Ca,L)), respectively, on SW reentry. METHODS: A two-dimensional epicardial ventricular muscle layer was created in rabbit hearts by cryoablation (n = 32), and action potential signals were analyzed by high-resolution optical mapping. RESULTS: During constant stimulation, pilsicainide (3-5 microM) caused a frequency-dependent decrease of conduction velocity (CV; by 20%-54% at 5 Hz) without affecting action potential duration (APD). Verapamil (3 microM) caused APD shortening (by 16% at 5 Hz) without affecting CV. Ventricular tachycardias (VTs) that were induced were more sustained in the presence of either pilsicainide or verapamil. The incidence of sustained VTs (>30 s)/all VTs per heart was 58% +/- 9% for 5 microM pilsicainide vs. 22% +/- 9% for controls and 62% +/- 10% for 3 microM verapamil vs. 22% +/- 8% for controls. The SWs with pilsicainide were characterized by slower rotation around longer functional block lines (FBLs), whereas those with verapamil were characterized by faster rotation around shorter FBLs. Combined application of 3 microM pilsicainide and 3 microM verapamil resulted in early termination of VTs (sustained VTs/all VTs per heart: 2% +/- 2% vs. 29% +/- 9% for controls); SWs showed extensive drift and decremental conduction, leading to their spontaneous annihilation. CONCLUSION: Blockade of either I(Na) or I(Ca,L) stabilizes SWs in a two-dimensional epicardial layer of rabbit ventricular myocardium to help their persistence, whereas blockade of both currents destabilizes SWs to facilitate their termination. PMID- 19303370 TI - Inappropriate implantable cardioverter-defibrillator therapy during exercise: what is the mechanism? PMID- 19303371 TI - Prevalence of long and short QT in a young population of 41,767 predominantly male Swiss conscripts. AB - BACKGROUND: Abnormally long and short QT intervals are recognized to be associated with an increased risk for life-threatening ventricular arrhythmias. It is therefore important to define the upper and lower border of the normal QT. OBJECTIVE: The aim of this study was to describe the normal distribution of the QT interval in a contemporary population of young conscripts and to define long and short limits of the QT interval. METHODS: In Switzerland, all young male citizens must undergo compulsory conscription for the Swiss Army at the age of 18 to 19 years. In every conscript, an electrocardiogram (ECG) is performed. Retrospectively, 41,767 consecutive ECGs of Swiss citizens who underwent conscription for the army between March 1, 2004, and July 31, 2006, were analyzed. RESULTS: The mean QTc Bazett interval was 394 +/- 22 ms. One percent of the conscripts had a Bazett QTc shorter than 347 ms, and one percent had a Bazett QTc longer than 445 ms, respectively. None of the subjects presented a QTc Bazett < 300 ms; the prevalence of a QTc Bazett < 320 ms was 0.02%. CONCLUSION: The present study shows the distribution of QT intervals in an unselected young population. Because none of the subjects presented a QTc < 300 ms, it may be concluded that the short QT syndrome is a very rare entity in the population of young male adults. PMID- 19303372 TI - Activation of 5-HT receptors in the periaqueductal gray attenuates the tachycardia evoked from dorsomedial hypothalamus. AB - Studies have shown that the dorsomedial hypothalamus (DMH) is a key region in the descending pathways mediating the cardiovascular response to emotional stress. We have recently demonstrated that the lateral/dorsolateral periaqueductal gray (l/dlPAG) is an important synaptic relay in mediating the tachycardic effect produced by activation of DMH neurons. This synaptic relay is mediated via NMDA excitatory amino acid receptors. In this study, our aim was to investigate, in conscious rats, whether activation of 5-Hydroxytriptamine 1A (5-HT(1A)) receptors in the l/dlPAG can attenuate the increases in heart rate and arterial pressure evoked by a) chemical activation of the DMH, b) air jet stress paradigm and c) chemical activation of l/dlPAG. Microinjections of the 5-HT(1A) receptor agonist, 8-OH-DPAT (1 nmol/100 nl), into the l/dlPAG reduced (by 62%) the increases in heart rate evoked by chemical activation of DMH neurons with the GABA(A) antagonist bicuculline methiodide (10 pmol/100 nl). The tachycardic and pressor responses evoked by air jet stress paradigm were also attenuated after treatment with 8-OH-DPAT in the l/dlPAG. The increases in heart rate and arterial pressure produced by microinjection of the excitatory amino acid receptor agonist, NMDA, into the l/dlPAG were largely reduced (by 94% and 73%, respectively) after treatment in the same region with 8-OH-DPAT. Taken together, our findings indicate that 5-HT(1A) receptors at the lateral dorsolateral PAG play a significant role in modulating the descending cardiovascular pathways from the dorsomedial hypothalamus and consequently the cardiovascular response to emotional stress. PMID- 19303373 TI - Synergistic decrease of DNA single-strand break repair rates in mouse neural cells lacking both Tdp1 and aprataxin. AB - Ataxia oculomotor apraxia-1 (AOA1) is an autosomal recessive neurodegenerative disease that results from mutations of aprataxin (APTX). APTX associates with the DNA single- and double-strand break repair machinery and is able to remove AMP from 5'-termini at DNA strand breaks in vitro. However, attempts to establish a DNA strand break repair defect in APTX-defective cells have proved conflicting and unclear. We reasoned that this may reflect that DNA strand breaks with 5'-AMP represent only a minor subset of breaks induced in cells, and/or the availability of alternative mechanisms for removing AMP from 5'-termini. Here, we have attempted to increase the dependency of chromosomal single- and double-strand break repair on aprataxin activity by slowing the rate of repair of 3'-termini in aprataxin-defective neural cells, thereby increasing the likelihood that the 5' termini at such breaks become adenylated and/or block alternative repair mechanisms. To do this, we generated a mouse model in which APTX is deleted together with tyrosyl DNA phosphodiesterase (TDP1), an enzyme that repairs 3' termini at a subset of single-strand breaks (SSBs), including those with 3' topoisomerase-1 (Top1) peptide. Notably, the global rate of repair of oxidative and alkylation-induced SSBs was significantly slower in Tdp1(-/-)/Aptx(-/-) double knockout quiescent mouse astrocytes compared with Tdp1(-/-) or Aptx(-/-) single knockouts. In contrast, camptothecin-induced Top1-SSBs accumulated to similar levels in Tdp1(-/-) and Tdp1(-/-)/Aptx(-/-) double knockout astrocytes. Finally, we failed to identify a measurable defect in double-strand break repair in Tdp1(-/-), Aptx(-/-) or Tdp1(-/-)/Aptx(-/-) astrocytes. These data provide direct evidence for a requirement for aprataxin during chromosomal single-strand break repair in primary neural cells lacking Tdp1. PMID- 19303374 TI - Co-morbidity of cystic fibrosis and celiac disease in Scandinavian cystic fibrosis patients. AB - BACKGROUND: The co-morbidity of cystic fibrosis (CF) and celiac disease (CD) has been reported sporadically since the 1960s. To our knowledge, this is the first time a systematic screening is performed in a large cohort of CF patients. METHODS: Transglutaminase-IgA (TGA), endomysium-IgA (EMA) and total IgA in serum were measured in 790 CF patients (48% females, 86% with pancreatic insufficiency). Six patients were diagnosed with CD prior to the study, all receiving a gluten-free diet. Patients with elevated TGA (>50 Units/mL) and a positive EMA test were offered a gastroscopy obtaining mucosal biopsies from the duodenum. RESULTS: Four new cases of CD were diagnosed. Two additional patients had positive serological tests, but normal biopsies. In total, 10 cases of CD (1.2%, 1:83) indicate a prevalence rate about three times higher than the general prevalence of CD in Norway and Sweden. No CD patients were detected in the Danish CF cohort. Patients diagnosed with untreated CD reported symptoms typical of both CF and CD (poor weight gain, loose and/or fatty stools, fatigue, irritability, abdominal pain). They improved after introduction of a gluten-free diet. CONCLUSIONS: Systematic screening for CD in a Scandinavian cohort of CF patients revealed a higher prevalence of CD than in the general population. Clinical signs of CD are difficult to differentiate from CF with malabsorption, and patients may go undiagnosed for a long time. In a population where CD is common we recommend screening for CD in patients with CF. PMID- 19303375 TI - Hyperkeratosis of nipple skin during sorafenib treatment. PMID- 19303376 TI - Predicting liver failure following major hepatectomy. AB - Pre-operative determination of the risk of liver dysfunction has come under criticism with regards to its usefulness in clinical practice. Opinion is split between centres which use such tests uniformly on all patients and those where clinical judgment alone is used. Published data would not suggest any difference in mortality, morbidity or liver failure rates between these groups. This review outlines and presents the evidence for pre-operative quantification of functional liver remnant volume. PMID- 19303377 TI - Role of surface charge and wettability on early stage mineralization and bone cell-materials interactions of polarized hydroxyapatite. AB - Our objective was to determine the role of surface charge and wettability on early stage mineralization as well as bone cell adhesion and proliferation on polarized HAp surface. To estimate the surface wettability, contact angles were measured in water, simulated body fluid (SBF) and Dulbecco's modified Eagle's medium/nutrient mixture F-12 Ham (DMEM). Experimental results show that HAp surface wettability and surface energy can be tailored by inducing surface charge without introducing any volumetric effects in the material. Increasing the surface charge increased the wettability and also the energy of HAp surfaces in all tested media. A maximum surface energy of 49.47+/-3.76mJ/m(2) was estimated for positively charged HAp surfaces polarized at 400(o)C. The in vitro bioactivity of polarized HAp samples was evaluated by soaking in SBF and DMEM (cell media). Cell-materials interaction was studied by culturing with human fetal osteoblast cells (hFOB). In vitro results show that tailoring the combined effect of wettability and charge polarity on the HAp surface enable differential binding of inorganic ions (e.g., Ca(2+), Cl(-), Na(+), HCO(3)(-) etc) and organic cell adhesive proteins (e.g., fibronectin, vitronectin etc) with different surface properties, which results in accelerated or decelerated mineralization as well as cell adhesion and proliferation on polarized HAp surface. PMID- 19303378 TI - Development of photocrosslinked methylcellulose hydrogels for soft tissue reconstruction. AB - A variety of materials have been used as fillers for soft tissue augmentation. In this study, methylcellulose (MC), a water-soluble derivative of the polysaccharide cellulose, was modified with functional methacrylate groups and photocrosslinked to produce hydrogels for potential application in plastic and reconstructive surgery. Purified macromer (5% theoretical modification, 2.3% actual) was resuspended in 0.05wt.% of the photoinitiator, 2-methyl-1-[4 (hydroxyethoxy)phenyl]-2-methyl-1-propanone, cast into custom molds, and exposed to long-wavelength UV light for 10min to form gels. Photocrosslinked MC hydrogels at varying weight/volume percentages displayed equilibrium weight swelling ratios (wet weight/dry weight) and elastic moduli of 30+/-3 to 17+/-2 and 8.48+/-0.25kPa to 23.21+/-1.55kPa, respectively, demonstrating the formation of stable gels with tunable properties. Human dermal fibroblasts grown in the presence of MC hydrogels in vitro exhibited no significant changes in cell viability after 5days of co-culture, indicating that the materials are non-cytotoxic. Higher weight percentage MC hydrogels (6%) implanted subcutaneously in CD-1 mice maintained their integrity and original dimensions after 80days in vivo, eliciting a mild inflammatory response with no observed inflammatory exudate, minimal vascular infiltration and thin translucent fibrous capsule formation of approximately 50microm in thickness. Taken together, the material and biological properties of photocrosslinked MC hydrogels suggest that they may be of use in soft tissue reconstruction. PMID- 19303379 TI - Damage control surgery. AB - The principles of trauma surgery have evolved during the past 20 years; from initial aggressive, definitive management of all surgical injuries in the traumatised patient to an abbreviated laparotomy, secondary correction of abnormal physiological parameters and then planned definitive re-exploration; the damage control sequence. PMID- 19303380 TI - Enterovirus infections in neonates. AB - Enteroviruses, which include echoviruses, coxsackie A and B viruses, polioviruses and the 'numbered' enteroviruses, are among the most common viruses causing disease in humans. A large proportion of enteroviral infections occur in neonates and infants. There is a wide spectrum of clinical manifestations that can be caused by enterovirus infection with varying degrees of severity. In the neonatal age group, enteroviral infections are associated with significant morbidity and mortality, particularly when infection occurs antenatally. This review provides a detailed overview of the epidemiology and clinical features of enterovirus infections in the neonatal period. In addition, laboratory features and diagnostic investigations are discussed. A review of the currently available data for prophylactic and therapeutic interventions, including antiviral therapy, is also presented. PMID- 19303381 TI - Samosa dressing in hand surgery. PMID- 19303382 TI - Home schooling of NK cells. AB - In this issue of Immunity, Chalifour et al. (2009) show that for natural killer (NK) cells to achieve their full effector potential, NK inhibitory receptors must developmentally interact with MHC-I ligands expressed in cis. PMID- 19303383 TI - Millipede-like lymphocyte crawling: feeling the way with filopodia. AB - In this issue of Immunity, Shulman et al. (2009) describe shear-facilitated chemokine-induced adhesive filopodia on crawling T lymphocytes that scan the endothelial surface for potential sites of transendothelial migration. These filopodia facilitate a novel millipede-like mode of lymphocyte locomotion over endothelial cells prior to extravasation. PMID- 19303384 TI - B cell multitasking is required to control nematode infection. AB - In this issue of Immunity, Wojciechowski et al. (2009) demonstrate that in addition to producing antibodies, B cells play pivotal roles in specific-antigen presentation and cytokine production for optimal T helper 2 responses required for protection against Heligmosomoides polygyrus. PMID- 19303385 TI - Live from the liver: hepatocyte IL-7. AB - Interleukin 7 (IL-7), which is required for T cell survival, was previously found in lymphoid tissues. In this issue of Immunity, Sawa et al. (2009) have identified the liver as a new source of IL-7. PMID- 19303386 TI - Arming Treg cells at the inflammatory site. AB - In this issue of Immunity, Zhang et al. (2009) propose that Treg cells, in order to efficiently control alloimmune response, need to be educated first in the target tissue before entering the draining lymph node. PMID- 19303388 TI - T cell-mediated autoimmune disease due to low-affinity crossreactivity to common microbial peptides. AB - Environmental factors account for 75% of the risk of developing multiple sclerosis (MS). Numerous infections have been suspected as environmental disease triggers, but none of them has consistently been incriminated, and it is unclear how so many different infections may play a role. We show that a microbial peptide, common to several major classes of bacteria, can induce MS-like disease in humanized mice by crossreacting with a T cell receptor (TCR) that also recognizes a peptide from myelin basic protein, a candidate MS autoantigen. Structural analysis demonstrates this crossreactivity is due to structural mimicry of a binding hotspot shared by self and microbial antigens, rather than to degenerate TCR recognition. Biophysical studies reveal that the autoreactive TCR binding affinity is markedly lower for the microbial (mimicry) peptide than for the autoantigenic peptide. Thus, these data suggest a possible explanation for the difficulty in incriminating individual infections in the development of MS. PMID- 19303387 TI - Follicular helper T cells: lineage and location. AB - Follicular helper T (Tfh) cells are the class of effector T helper cells that regulates the step-wise development of antigen-specific B cell immunity in vivo. Deployment of CXCR5+ Tfh cells to B cell zones of lymphoid tissues and stable cognate interactions with B cells are central to the delivery of antigen-specific Tfh cell function. Here, we review recent advances that have helped to unravel distinctive elements of developmental programming for Tfh cells and unique effector Tfh cell functions focused on antigen-primed B cells. Understanding the regulatory functions of Tfh cells in the germinal center and the subsequent regulation of memory B cell responses to antigen recall represent the frontiers of this research area with the potential to alter fundamentally the design of future vaccines. PMID- 19303389 TI - The lymphotoxin LTalpha(1)beta(2) controls postnatal and adult spleen marginal sinus vascular structure and function. AB - The lymphotoxin LTalpha(1)beta(2) supports the development and maintenance of several aspects of spleen structure, but its significance for marginal sinus (MS) vascular organization is unclear. We showed here that, in early postnatal lymphotoxin-deficient mice, the developing Flk-1+ white pulp vessels failed to organize or upregulate MAdCAM-1, leading to altered spatial rearrangement of both the white pulp endothelial cells and the smooth muscle actin-expressing cells. In vitro, MAdCAM-1 directed the reorganization of LTbeta receptor+ endothelial cells grown on Matrigel. LTalpha(1)beta(2) also regulated the maintenance of both MAdCAM-1 expression and mature MS structure in adult mice, contributing importantly to normal trafficking of CD11b+ cells in response to bacterial antigens. Together, our studies demonstrate that LTalpha(1)beta(2) and LTbeta receptor signals control proper development and maintenance of the mature MS structure and implicate MAdCAM-1 in the structuring of the MS endothelial cells that is important for the movement of immune cells within the spleen. PMID- 19303390 TI - Regulatory T cells sequentially migrate from inflamed tissues to draining lymph nodes to suppress the alloimmune response. AB - To determine the site and mechanism of suppression by regulatory T (Treg) cells, we investigated their migration and function in an islet allograft model. Treg cells first migrated from blood to the inflamed allograft where they were essential for the suppression of alloimmunity. This process was dependent on the chemokine receptors CCR2, CCR4, and CCR5 and P- and E-selectin ligands. In the allograft, Treg cells were activated and subsequently migrated to the draining lymph nodes (dLNs) in a CCR2, CCR5, and CCR7 fashion; this movement was essential for optimal suppression. Treg cells inhibited dendritic cell migration in a TGF beta and IL-10 dependent fashion and suppressed antigen-specific T effector cell migration, accumulation, and proliferation in dLNs and allografts. These results showed that sequential migration from blood to the target tissue and to dLNs is required for Treg cells to differentiate and execute fully their suppressive function. PMID- 19303391 TI - Comparative removal of solvent and detergent viral inactivating agents from human intravenous immunoglobulin G preparations using SDR HyperD and C18 sorbents. AB - The capacity of hydrophobic octadecyl (C18) and SDR HyperD materials to remove the combination of 1% (v/v) solvent (tri-n-butyl phosphate, TnBP) with 1% (v/v) nonionic detergents (Triton X-100 and Triton X-45) used for viral inactivation of plasma-derived polyvalent intravenous immunoglobulin G (IVIG) preparation has been evaluated. Efficient removal of TnBP (<10 ppm in IVIG preparation) was found at ratios of 0.5 g of C18/7 ml of IVIG and 0.22 g of dry SDR HyperD/7 ml of IVIG. Binding capacities of TnBP were greater than 140 mg/g of C18 and greater than 318 mg/g of dry SDR HyperD. Complete removal of Triton X-45 (<2 ppm) was obtained at ratios of 1g of C18/7 ml of IVIG and 0.44 g of dry SDR HyperD/7 ml of IVIG or above, corresponding to binding capacities in excess of 70 mg/g of C18 and in excess of 159 mg/g of dry SDR HyperD. Residual Triton X-100 was less than 30 ppm at a ratio of 4 g/14 ml of immunoglobulin G (IgG) for the C18 sorbent. Triton X 100 was less than 10 ppm when using SDR HyperD at a ratio of 0.66 g/7 ml of IgG, corresponding to a binding capacity of approximately 106 mg of Triton X-100/g of dry SDR HyperD. Good recoveries of IVIG were achieved in the effluent from both sorbents. PMID- 19303392 TI - A solution for stripping antibodies from polyvinylidene fluoride immunoblots for multiple reprobing. AB - Available protocols for stripping antibodies from immunoblots involve the use of sodium dodecyl sulfate (SDS) or low-pH buffers. SDS was shown to remove transferred proteins from membranes, and low-pH buffer was shown to inefficiently strip off antibodies. A solution containing 6M guanidine hydrochloride, 0.2% nondenaturing detergent, and a reducing agent can rapidly strip off tightly bound antibodies from aged polyvinylidene fluoride (PVDF) immunoblots at room temperature without removing significant amounts of transferred protein. PMID- 19303393 TI - pTOM, a vector for expressing two proteins in mammalian cells. AB - We developed a pTOM construct that can express two proteins in a cell. Using pTOM ensures that two polypeptide-encoding nucleotide sequences are simultaneously transfected into the same cell. The ability to simultaneously express two separate polypeptide-encoding nucleotide sequences from the same vector in the same cell allows the user to determine the relationship between the two proteins. Another advantage is that one of the proteins can be used as a transfection marker/selector. The vector contains multiple cloning sites for insertions of polypeptide-encoding nucleotide sequences. Positive clones can be easily selected when performing cloning using ampicillin. Overall, this vector provides a convenient way to express dual proteins in a single mammalian cell. PMID- 19303394 TI - Bone morphogenetic protein-7 and Gremlin: New emerging therapeutic targets for diabetic nephropathy. AB - Specific therapies of diabetic nephropathy (DN) are not available, and current treatment strategies are limited to management of blood glucose levels and control of hypertension. The re-activation of developmental programs in DN suggests new potential therapeutic targets. Bone morphogenetic protein-7 (BMP-7) and its antagonist, Gremlin is revealed to be involved in renal development and diabetic nephropathy. This article reviews the changes of BMP-7 and Gremlin in diabetic kidney, the protective effects on diabetic nephropathy when targeting BMP-7 and Gremlin, and the possible mechanism. The reorganization of the re activation of Gremlin and BMP-7 in diabetic kidney had shed light on the identification of novel therapeutic targets for DN. PMID- 19303395 TI - Identification and characterization of flavonoids as sialyltransferase inhibitors. AB - Sialyltransferases biosynthesize sialyl-glycoconjugates involved in many biological and pathological processes. We investigated and characterized synthetic flavonoid derivatives as sialyltransferase inhibitors. We first examined 54 compounds by solid-phase enzyme assay using beta-galactoside alpha2,6 sialyltransferase 1 (ST6Gal I) and beta-galactoside alpha2,3-sialyltransferase. Several compounds inhibited sialyltransferase enzyme activity regardless of sialyl-linkage reactions. Among them, two compounds showed inhibitory activity against ST6Gal I with IC(50) values less than 10 microM. Three characteristic features of flavonoids were determined by structure-inhibitory activity relationships. First, a double bond between C2-C3 linkages is required for the activity. Second, increasing hydrophilic properties on the B-ring markedly augmented the inhibitory effect. Third, a hydrophobic functional group introduced on the hydroxyl groups of the A-ring enhanced the inhibitory activity. Kinetic analysis using human ST6Gal I indicated a mixed inhibition mechanism of the compounds. In conclusion, the flavonoids identified could be applied for control of cellular expression of sialic acid. PMID- 19303396 TI - NKG2D and CD94 bind to multimeric alpha2,3-linked N-acetylneuraminic acid. AB - Killer lectin-like receptors on natural killer cells mediate cytotoxicity through glycans on target cells including the sialyl Lewis X antigen (sLeX). We investigated whether NK group 2D (NKG2D) and CD94 can bind to sialylated N-linked glycans, using recombinant glutathione S-transferase-fused extracellular lectin like domains of NKG2D (rNKG2Dlec) and CD94 (rCD94lec). Both rNKG2Dlec and rCD94lec bound to plates coated with high-sLeX-expressing transferrin secreted by HepG2 cells (HepTF). The binding of rNKG2Dlec and rCD94lec to HepTF was markedly suppressed by treatment of HepTF with neuraminidase and in the presence of N acetylneuraminic acid. Moreover, rNKG2Dlec and rCD94lec bound to alpha2,3 sialylated human alpha(1)-acid glycoprotein (AGP) but not to alpha2,6-sialylated AGP. Mutagenesis revealed that (152)Y of NKG2D and (144)F and (160)N of CD94 were critical for HepTF binding. This is the first report that NKG2D and CD94 bind to alpha2,3-sialylated but not to alpha2,6-sialylated multi-antennary N-glycans. PMID- 19303397 TI - Different catalytic properties of two highly homologous triosephosphate isomerase monomers. AB - It is assumed that amino acid sequence differences in highly homologous enzymes would be found at the peripheral level, subtle changes that would not necessarily affect catalysis. Here, we demonstrate that, using the same set of mutations at the level of the interface loop 3, the activity of a triosephosphate isomerase monomeric enzyme is ten times higher than that of a homologous enzyme with 74% identity and 86% similarity, whereas the activity of the native, dimeric enzymes is essentially the same. This is an example of how the dimeric biological unit evolved to compensate for the intrinsic differences found at the monomeric species level. Biophysical techniques of size exclusion chromatography, dynamic light scattering, X-ray crystallography, fluorescence and circular dichroism, as well as denaturation/renaturation assays with guanidinium hydrochloride and ANS binding, allowed us to fully characterize the properties of the new monomer. PMID- 19303398 TI - BetaPIX and GIT1 regulate HGF-induced lamellipodia formation and WAVE2 transport. AB - Formation of lamellipodia is the first step during cell migration, and involves actin reassembly at the leading edge of migrating cells through the membrane transport of WAVE2. However, the factors that regulate WAVE2 transport to the cell periphery for initiating lamellipodia formation have not been elucidated. We report here that in human breast cancer MDA-MB-231 cells, the hepatocyte growth factor (HGF) induced the association between the constitutive complex of betaPIX and GIT1 with WAVE2, which was concomitant with the induction of lamellipodia formation and WAVE2 transport. Although depletion of betaPIX by RNA interference abrogated the HGF-induced WAVE2 transport and lamellipodia formation, GIT1 depletion caused HGF-independent WAVE2 transport and lamellipodia formation. Collectively, we suggest that betaPIX releases cells from the GIT1-mediated suppression of HGF-independent responses and recruits GIT1 to WAVE2, thereby facilitating HGF-induced WAVE2 transport and lamellipodia formation. PMID- 19303399 TI - An anti-apoptotic peptide improves survival in lethal total body irradiation. AB - Cell penetrating peptides (CPPs) have been used to deliver the anti-apoptotic Bcl xL-derived BH4 peptide to prevent injury-induced apoptosis both in vitro and in vivo. Here we demonstrate that the nuclear localization sequence (NLS) from the SV40 large T antigen has favorable properties for BH4 domain delivery to lymphocytes compared to sequences based on the HIV-1 TAT sequence. While both TAT BH4 and NLS-BH4 protected primary human mononuclear cells from radiation-induced apoptotic cell death, TAT-BH4 caused persistent membrane damage and even cell death at the highest concentrations tested (5-10 microM) and correlated with in vivo toxicity as intravenous administration of TAT-BH4 caused rapid death. The NLS-BH4 peptide has significantly attenuated toxicity compared to TAT-BH4 and we established a dosing regimen of NLS-BH4 that conferred a significant survival advantage in a post-exposure treatment model of LD90 total body irradiation. PMID- 19303400 TI - FAK mediates the inhibition of glioma cell migration by truncated 24 kDa FGF-2. AB - A truncated form of 24kDa FGF-2 consisting of 86 NH(2)-terminal amino acids (ATE+31) inhibits cell migration in vitro and tumor development and angiogenesis in vivo. Focal adhesion kinase (FAK) is phosphorylated on tyrosine and serine sites after cell stimulation by growth factors. This study examined the effect of ATE+31 on FAK phosphorylation in human glioma cells. FAK and Pyk phosphorylation were evaluated at serines known to be involved with cell migration. We demonstrated that ATE+31 at 3 x 10(-11)M decreases phosphorylation levels of Tyr(407)-FAK and Ser(732)-FAK in the presence of platelet-derived growth factor (PDGF), that ATE+31 in the presence of PDGF alters the distribution of FAK and other phosphotyrosine proteins in the adhesion contacts, and that ATE+31 in the presence of PDGF has no effect on the activation of Pyk2. These data suggest that the inhibition of cell migration by ATE+31 occurs via Tyr(407)-FAK and Ser(732) FAK. PMID- 19303401 TI - Characterization of arginase 1 gene polymorphisms in the Algerian population and association with blood pressure. AB - OBJECTIVES: (i) To characterize the polymorphism of arginase 1 (ARG1), a new candidate gene in coronary heart disease (CHD), in the Algerian population; (ii) To evaluate the effect of common ARG1 single nucleotide polymorphisms (SNPs) on blood pressure (BP) values; and (iii) To compare the data with those previously obtained in French populations. METHODS: Eleven ARG1 SNPs selected from databases were characterized in a representative sample of 117 Algerian and 92 French males free of CHD. Relevant SNPs for association studies with BP were selected on the basis of their allele frequencies and pairwise linkage disequilibrium. RESULTS: ARG1 allele frequencies and haplotype distribution significantly differed between Algerian and French subjects. The rs2781667C/T polymorphism was associated with decreased systolic BP in Algerian subjects. This association contrasted with previous data we reported in the French population. The discrepancy would be explained by the difference in haplotype distribution between Algerian and French subjects. CONCLUSION: These data support the role of ARG1 in vascular pathophysiology, but the functional mutations remain to be identified. PMID- 19303402 TI - Performance characteristics of a no-pretreatment, random access sirolimus assay for the Dimension RxL clinical chemistry system. AB - OBJECTIVES: Current therapeutic drug monitoring methods for sirolimus require a manual pre-treatment step and batch analysis. We describe and validate a no pretreatment, random access sirolimus assay for the Dimension RxL clinical chemistry system from Siemens Healthcare Diagnostics Inc. DESIGN AND METHODS: Whole blood samples from renal transplant patients prescribed sirolimus were analyzed by the LC-MS/MS reference method, Abbott IMx and Dimension RxL methods in accordance with CLSI recommendations. RESULTS: The Dimension sirolimus assay had a functional sensitivity of 2.0 ng/mL and repeatability and within-laboratory imprecision less than 12.6% at 3 ng/mL and less than 5% at 11-12 ng/mL. Least squares linear regression demonstrated the following relationships: RxL=1.20(LC MS/MS) - 0.70, r=0.95 and RxL=1.33(IMx) - 0.75, r=0.96. CONCLUSIONS: The Dimension sirolimus assay correlates well with the LC-MS/MS reference and IMx immunoassay methods and has the advantage of random access analysis without a manual pre-treatment step. PMID- 19303403 TI - High-throughput single cell arrays as a novel tool in biopreservation. AB - Microwell array cytometry is a novel high-throughput experimental technique that makes it possible to correlate pre-stress cell phenotypes and post-stress outcomes with single cell resolution. Because the cells are seeded in a high density grid of cell-sized microwells, thousands of individual cells can be tracked and imaged through manipulations as extreme as freezing or drying. Unlike flow cytometry, measurements can be made at multiple time points for the same set of cells. Unlike conventional image cytometry, image analysis is greatly simplified by arranging the cells in a spatially defined pattern and physically separating them from one another. To demonstrate the utility of microwell array cytometry in the field of biopreservation, we have used it to investigate the role of mitochondrial membrane potential in the cryopreservation of primary hepatocytes. Even with optimized cryopreservation protocols, the stress of freezing almost always leads to dysfunction or death in part of the cell population. To a large extent, cell fate is dominated by the stochastic nature of ice crystal nucleation, membrane rupture, and other biophysical processes, but natural variation in the initial cell population almost certainly plays an important and under-studied role. Understanding why some cells in a population are more likely to survive preservation will be invaluable for the development of new approaches to improve preservation yields. For this paper, primary hepatocytes were seeded in microwell array devices, imaged using the mitochondrial dyes Rh123 or JC-1, cryopreserved for up to a week, rapidly thawed, and checked for viability after a short recovery period. Cells with a high mitochondrial membrane potential before freezing were significantly less likely to survive the freezing process, though the difference in short term viability was fairly small. The results demonstrate that intrinsic cell factors do play an important role in cryopreservation survival, even in the short term where extrinsic biophysical factors would be expected to dominate. We believe that microwell array cytometry will be an important tool for a wide range of studies in biopreservation and stress biology. PMID- 19303404 TI - Phospholipase C mediated Ca2+ signals in murine urinary bladder smooth muscle. AB - Muscarinic stimulation of urinary bladder induces contraction via an increase in intracellular Ca(2+) concentration that results from Ca(2+) influx through Ca(2+) channels and/or IP(3)-mediated Ca(2+) release controlled by phospholipase C (PLC) signalling. The significance of PLC/IP(3) signalling in this cascade has recently been questioned because PLC inhibitors were without effect on carbachol-induced contractions in detrusor muscle strips. However, PLC/IP(3)-mediated Ca(2+) release was clearly observed in recordings of Ca(2+) signals in isolated myocytes. Therefore, we investigated the presence of PLC/IP(3)-dependent Ca(2+) release by directly monitoring Ca(2+) signals in intact detrusor muscle strips. Concomitant Ca(2+) signals from Ca(2+) channel activity were eliminated by the Ca(2+) channel antagonist isradipine (3 microM) or by the use of muscles from Ca(v)1.2 channel-deficient (SMACKO) mice. In absence of Ca(2+) channel activity, carbachol elicited contractions and Ca(2+) signals in muscles from wild type and SMACKO mice that were inhibited by the PLC inhibitor U73122 (10 microM). The results show that PLC/IP(3)-dependent Ca(2+) release is activated by stimulation with carbachol in urinary bladder smooth muscle but has a minor contribution to overall carbachol-induced Ca(2+) signals. PMID- 19303405 TI - In vivo normobaric hyperoxia preconditioning induces different degrees of antioxidant enzymes activities in rat brain tissue. AB - Recent studies suggest that intermittent and prolonged normobaric hyperoxia results in ischemic tolerance to reduce ischemia brain injury. In this research attempts were made to see the changes in antioxidant enzyme activities following prolonged and intermittent normobaric hyperoxia preconditioning. Rats were divided into four experimental groups, each of 21 animals. The first two were exposed to 95% inspired normobaric hyperoxia for 4 h/day for 6 consecutive days (intermittent normobaric hyperoxia) or for 24 h continuous (prolonged normobaric hyperoxia). The second two groups acted as controls, and were exposed to 21% oxygen in the same chamber. Each main group was subdivided to middle cerebral artery occlusion-operated, sham-operated (without middle cerebral artery occlusion), and intact (without any surgery) subgroups. After 24 h, middle cerebral artery occlusion-operated subgroups were subjected to 60 min of right middle cerebral artery occlusion. After 24 h reperfusion, neurologic deficit score, infarct volume were measured in middle cerebral artery occlusion-operated subgroups. Antioxidant enzyme activities were assessed in sham-operated and intact subgroups. Preconditioning with prolonged and intermittent normobaric hyperoxia decreased neurologic deficit score and infarct volume, and increased antioxidant enzyme activities (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) significantly. Although further studies are needed to clarify the mechanisms of ischemic tolerance, the intermittent and prolonged normobaric hyperoxia seems to partly exert their effects via increase antioxidant enzymes activities. PMID- 19303406 TI - Resveratrol prevents memory deficits and the increase in acetylcholinesterase activity in streptozotocin-induced diabetic rats. AB - The objective of the present study was to investigate the effect of the administration of resveratrol (RV) on memory and on acetylcholinesterase (AChE) activity in the cerebral cortex, hippocampus, striatum, hypothalamus, cerebellum and blood in streptozotocin-induced diabetic rats. The animals were divided into six groups (n=6-13): Control/saline; Control/RV 10 mg/kg; Control/RV 20 mg/kg; Diabetic/saline; Diabetic/RV 10 mg/kg; Diabetic/RV 20 mg/kg. One day after 30 days of treatment with resveratrol the animals were submitted to behavioral tests and then submitted to euthanasia and the brain structures and blood were collected. The results showed a decrease in step-down latency in diabetic/saline group. Resveratrol (10 and 20 mg/kg) prevented the impairment of memory induced by diabetes. In the open field test, no significant differences were observed between the groups. In relation to AChE activity, a significant increase in diabetic/saline group (P<0.05) was observed in all brain structures compared to control/saline group. However, AChE activity decreased significantly in control/RV10 and control/RV20 (P<0.05) groups in cerebral cortex, hippocampus and striatum, while no significant differences were observed in diabetic/RV10 and diabetic/RV20 groups in all brain structures compared to control/saline group. Blood AChE activity increased significantly in diabetic/saline group (P<0.05) decreased in control/RV10, control/RV20 and diabetic/RV20 groups (P<0.05) compared to control/saline group. In conclusion, the present findings showed that treatment with resveratrol prevents the increase in AChE activity and consequently memory impairment in diabetic rats, demonstrating that this compound can modulate cholinergic neurotransmission and consequently improve cognition. PMID- 19303407 TI - Relaxation of tracheal smooth muscle independent on functional epithelium cells induced by lidocaine, bupivacaine and isomers in rats. AB - Lidocaine is a local anesthetic which has been used to protect spasm reaction during tracheal intubation and bronchoscopy. We compared the potency of lidocaine, bupivacaine (RS(+/-)-bupivacaine) and isomers (S(-)-bupivacaine and R(+)-bupivacaine) to promote relaxation of tracheal smooth muscle. Relaxation of airways smooth muscle can be dependent on the release of relaxing factors by epithelium such as prostanoids and nitric oxide (NO). Possible mechanisms involved in the tracheal smooth muscle relaxation induced by these local anesthetics were evaluated in preparation in which the epithelium layer was intact or denuded. Bupivacaine and its isomers were approximately six to eleven fold more potent than lidocaine to promote relaxation on acetylcholine-induced contraction in tracheal rings. The concentration of lidocaine, RS(+/-) bupivacaine, S(-)-bupivacaine and R(+)-bupivacaine necessary to produce a 50% reduction of maximal contraction to acetylcholine (IC(50)) in tracheal rings with intact epithelium was 1.25+/-0.01, 0.11+/-0.01, 0.15+/-0.01, 0.19+/-0.01 mM, respectively. Removal of epithelium or exposure to N(G)-nitro-L-arginine methyl ester, indomethacin did not alter the IC(50). However, calcium influx of depolarized tracheal smooth muscle was inhibited by lidocaine, bupivacaine and isomers. S(-)-bupivacaine reduced by 78.8+/-7.4% the calcium influx followed by RS(+/-)-bupivacaine (41.8+/-6.7%) and R(+)-bupivacaine (25.6+/-9.5%). In conclusion, local anesthetic action was stereoselective and partially dependent on blockade of Ca(2+) influx to muscular cells. The isomer S(-)-bupivacaine is more potent and less toxic which could represent a valuable clinical advantage to use as broncholitic agent. PMID- 19303408 TI - Pathogenesis gene families in the common minimal genome of Staphylococcus aureus are hypervariable. AB - Staphylococcus aureus is a versatile pathogen that shows high levels of inter strain genetic variability and positive evolution in certain pathogenesis-related genes. Apart from gene content differences, variability in shared genes may affect pathogenicity. Studying such variability requires that the common minimal genome (CMG) be identified. In this study, we have surveyed the CMG of S. aureus with respect to variability amongst orthologous family members, and determined that genes involved in pathogenesis preferentially accumulate variations. A negative correlation between variability of genes and their evolution was found, suggesting a preservation of host-specific function while exhibiting sequence diversity. Variation in key pathogenesis genes in S. aureus might predispose them to functional modulation, thereby playing an important role in evasion of host immunity. PMID- 19303409 TI - Micellar lipid composition profoundly affects LXR-dependent cholesterol transport across CaCo2 cells. AB - Intraluminal phospholipids affect micellar solubilization and absorption of cholesterol. We here study cholesterol transport from taurocholate-phospholipid cholesterol micelles to CaCo2 cells, and associated effects on ABC-A1 mediated cholesterol efflux. Micellar incorporation of egg-yolk-phosphatidylcholine markedly increased apical retention of the sterol with decreased expression of ABC-A1, an effect that is prevented by synthetic liver X receptor (LXR) or retinoid X receptor (RXR) agonists. On the other hand, incorporation of lyso phosphatidylcholine (LysoPC) increased ABC-A1-HDL-dependent basolateral cholesterol efflux, an effect that is abated when LXR is silenced. Thus, the modulation of cholesterol metabolism via intraluminal phospholipids is related to the activity of the oxysterol nuclear receptor LXR. PMID- 19303410 TI - Interaction between Sox proteins of two physiologically distinct bacteria and a new protein involved in thiosulfate oxidation. AB - Organisms using the thiosulfate-oxidizing Sox enzyme system fall into two groups: group 1 forms sulfur globules as intermediates (Allochromatium vinosum), group 2 does not (Paracoccus pantotrophus). While several components of their Sox systems are quite similar, i.e. the proteins SoxXA, SoxYZ and SoxB, they differ by Sox(CD)(2) which is absent in sulfur globule-forming organisms. Still, the respective enzymes are partly exchangeable in vitro: P. pantotrophus Sox enzymes work productively with A. vinosum SoxYZ whereas A. vinosum SoxB does not cooperate with the P. pantotrophus enzymes. Furthermore, A. vinosum SoxL, a rhodanese-like protein encoded immediately downstream of soxXAK, appears to play an important role in recycling SoxYZ as it increases thiosulfate depletion velocity in vitro without increasing the electron yield. PMID- 19303411 TI - The N-terminal fragment of Reelin is generated after endocytosis and released through the pathway regulated by Rab11. AB - Reelin is a large secreted glycoprotein essential for brain formation, but its trafficking and function at the molecular level remain incompletely understood. After binding to its receptor, Reelin is internalized by endocytosis. Here we show that internalized Reelin is subject to specific proteolysis within the cell and its N-terminal fragment is re-secreted. This re-secretion is inhibited by bafilomycin A(1) or by expression of a mutant of Rab11, a regulator of the recycling pathway. As the N-terminal fragment does not bind to Reelin receptor but has homology to F-spondin, its recycling may be involved in the regulation of extracellular matrix. PMID- 19303412 TI - Nm23-M5 mediates round and elongated spermatid survival by regulating GPX-5 levels. AB - Nucleoside diphosphate (NDP) kinases are involved in numerous regulatory processes associated with proliferation, development, and differentiation. Previously, we cloned a new member of the NDPK family from mouse, Nm23-M5, which encodes a 211-amino acid protein and has 86% identity to the human Nm23-H5 [Hwang, K.C., Ok, D.W., Hong, J.C., Kim, M.O. and Kim, J.H. (2003) Cloning, sequencing, and characterization of the murine Nm23-M5 gene during mouse spermatogenesis and spermiogenesis. Biochem. Biophys. Res. Commun. 306, 198-207]. To better understand Nm23-M5 function, we generated transgenic mice with reduced Nm23-M5 levels in vivo using a short hairpin RNA (shRNA) knock-down system. Nm23 M5 expression was markedly reduced, as indicated by Northern and Western blot analysis. Nm23-M5 shRNA transgenic mice exhibited reduced numbers of haploid cells. Furthermore, the antioxidant enzyme glutathione peroxidase 5 (GPX-5) is regulated by Nm23-M5 at the level of both expression and activity. These results reveal that expression of Nm23-M5 plays a critical role in spermiogenesis by increasing the cellular levels of GPX-5 to eliminate reactive oxygen species. PMID- 19303413 TI - Assembly of a chimeric respiratory chain from bovine heart submitochondrial particles and cytochrome bd terminal oxidase of Escherichia coli. AB - Cytochrome bd is a terminal quinol oxidase in Escherichia coli. Mitochondrial respiration is inhibited at cytochrome bc(1) (complex III) by myxothiazol. Mixing purified cytochrome bd oxidase with myxothiazol-inhibited bovine heart submitochondrial particles (SMP) restores up to 50% of the original rotenone sensitive NADH oxidase and succinate oxidase activities in the absence of exogenous ubiquinone analogs. Complex III bypassed respiration and is saturated at amounts of added cytochrome bd similar to that of other natural respiratory components in SMP. The cytochrome bd tightly binds to the mitochondrial membrane and operates as an intrinsic component of the chimeric respiratory chain. PMID- 19303414 TI - Sustained response of hepatitis B e antigen-negative patients 3 years after treatment with peginterferon alpha-2a. AB - BACKGROUND & AIMS: Patients with hepatitis B e antigen (HBeAg)-negative chronic hepatitis B treated with peginterferon alpha-2a with or without lamivudine achieve significantly higher 6-month posttreatment rates of response compared with those treated with lamivudine alone. The durability of 51 years old) had a statistically significant effect on any parameter (p> or =0.05). DISCUSSION: Although measurements of CSF flow dynamics parameter are sensitive to various factors (temporal and spatial resolutions of the MR sequence, circadian rhythms, age, sex, and MR equipment manufacturer), an optimized post-processing method enables reliable and reproducible values and ranges for normal subjects to be established. PMID- 19303480 TI - [MRI quantification of myocardial function, perfusion, and enhancement in patients with left-ventricular noncompaction]. AB - OBJECTIVE: Left ventricular noncompaction is a congenital malformation characterized by a myocardium organized into two layers, one compacted and one noncompacted. We aimed to quantify myocardial function, perfusion, and delayed enhancement using MRI in patients with left-ventricular noncompaction and to compare these results with those of normal patients. MATERIAL AND METHODS: We included 12 patients with a myocardial noncompaction / compaction ratio>2.3 at end-diastole in at least one segment apart from the apex and 12 healthy subjects matched for age and sex. We calculated the end-diastolic and end-systolic volumes, stroke volume, ejection fraction, cardiac output, myocardial volume and mass, end-diastolic thickness, and left-ventricular wall thickening and motion. From the delayed enhancement images, we obtained the volume and percentage of hyperenhanced myocardium. Student's t test was used to compare groups. RESULTS: We observed a statistically significant increase in end-diastolic and end systolic volumes in patients with left-ventricular noncompaction, as well as decreased ejection fraction, wall motion, and relative maximum upslope in segments 4, 9, and 10. No significant differences were found in delayed hyperenhancement. CONCLUSION: MRI quantification revealed decreased systolic cardiac function and decreased perfusion (lower relative maximum upslope) in the lower segments in patients with noncompaction. PMID- 19303481 TI - [Treating iatrogenic pneumothorax with small-bore chest tube]. AB - OBJECTIVE: To evaluate the feasibility of small-bore chest tube placement to treat pneumothorax caused by percutaneous procedures. MATERIAL AND METHODS: Between November 2004 and July 2006 we performed 151 interventional chest procedures (127 biopsies and 24 radiofrequency ablations) in 131 patients (25 women and 106 men; mean age, 63 years, range, 36-83 years). Fine needles (21-25 G) were used in 70 procedures and large core needles (14-20 G) were used in the remaining 81. Pneumothorax occurred in 16 diagnostic procedures (13 of these were treated with small-bore catheter placement) and in two radiofrequency procedures (both cases were treated with small-bore catheter placement). Chest tubes were placed immediately after pneumothorax occurred in all cases because the pneumothorax was greater than 20% or caused symptoms or occurred in patients with emphysema. RESULTS: Chest tubes were successfully placed without incidents in all cases and a Heimlich valve evacuated the air completely. Mean hospital stay was 43 hours (range, 24-72 hours). It was not necessary to place a larger-bore chest tube in any case. CONCLUSION: Small-bore chest tube placement is the treatment of choice for iatrogenic pneumothorax greater than 20% or less than 20% when symptomatic or occurring in patients with emphysema. The procedure is easy, effective, and well tolerated; furthermore, it shortens the hospital stay. The effectiveness of the procedure makes it possible to safely perform percutaneous procedures on patients with emphysema or difficult lesions and to finish an interventional procedure when pneumothorax occurs. PMID- 19303482 TI - [Unusual imaging findings of Ewing sarcoma in the childhood]. AB - OBJECTIVES: To review the cases of Ewing's sarcoma (ES) diagnosed in children at our hospital from 1995 to 2005 and to analyze uncommon imaging signs at diagnosis or during the course of the disease. MATERIAL AND METHODS: We reviewed the cases of 21 patients with ES (12 boys and 9 girls) diagnosed between the ages of 2 and 14 years. We analyzed the types of presentation, the imaging findings (plain films, CT, and MRI), and the patients' evolution. All cases were confirmed at histological study. RESULTS: The following uncommon manifestations were found in 11 patients: 2 vertebral, one cervical, and one dorsal ES that initially manifested as spinal cord compression and as a mediastinal mass, respectively; 2 sacral ES with ascending epidural masses; 1 mandibular ES; 1 costal ES with hemorrhagic cerebral metastases; 3 extraskeletal (thigh, buttocks, and pelvis minor) ES with bone metastases at diagnosis: the first two had spinal cord and nerve compression due to vertebral metastases; 2 ES had benign-appearing radiological findings (cortical lesion of the radius with osseous remodeling of the ulna and an expanding multilocular lesion of the fibula). CONCLUSIONS: Although ES is the second most common malignant bone tumor in children, its unusual imaging findings are less well known. More than half (52.4 %) of our last 21 patients presented some of these manifestations at diagnosis or during follow up, so radiologists should be aware of them and include ES in the prebiopsy diagnostic possibilities. PMID- 19303483 TI - [Treatment of an abdominal aorta aneurysms with endovascular stent grafts and embolizing materials: an experimental study]. AB - OBJECTIVE: To create an animal model to stabilize the aneurysmal sac and prevent type II leaks in the endovascular treatment of aneurysms of the abdominal aorta (AAA) by placing an endoprosthesis (stent-graft) and filling the aneurysmal sac. MATERIAL AND METHODS: AAA were experimentally induced in pigs and sheep; AAA were subsequently treated with endovascular stent grafts and stabilized in the same procedure by introducing ethibloc and onyx through a vascular catheter placed within the aneurysmal sac. All animals underwent aortography, and MR angiography and abdominal CT were performed on randomly selected animals. Histological studies included both macroscopic and microscopic examination. Two sheep were analyzed one year after the procedure. We evaluated the characteristics of the materials employed, their diffusion in the collateral arteries, and their efficacy in stabilizing the aneurysmal sac. RESULTS: The procedure was successfully carried out in all animals. Survival in the pigs was 100 %, whereas greater instability under anesthesia in the sheep led to 50 % perioperative mortality. Both ethibloc and onyx were considered easy to use and stable over time. CONCLUSION: The sac of an AAA treated with an endovascular stent graft and its collateral vessels can be treated in the same procedure with materials introduced through a catheter. PMID- 19303484 TI - [Percutaneous fixation of unstable pelvic ring disruption under CT guidance]. AB - CT-guided percutaneous osteosynthesis of posterior disruption of an unstable pelvic ring is an alternative to the fluoroscopically guided procedure or to open reduction and internal fixation (ORIF), which is the technique of choice for unstable pelvic fracture with disruption of the sacroiliac joint or fracture of the sacral wing. However, ORIF has a high rate of morbidity and mortality, and the CT-guided procedure involves less blood loss and has a lower rate of infectious and postoperative complications. CT guidance of the percutaneous technique also has significant advantages over fluoroscopic guidance. The critical point is the placement of the guidewire inside the sacral body through the external surface of the iliac wing and of the fracture line or sacroiliac diastasis. We describe and illustrate the procedure through three cases, two sacroiliac articular disruptions and a fracture of the sacral wing. PMID- 19303485 TI - [Embolotherapy of pulmonary arteriovenous malformations: long-term clinical and radiological follow-up]. AB - Pulmonary arteriovenous malformations (PAVM) are abnormal communications between the pulmonary arteries and pulmonary veins. The large majority are congenital and are commonly associated with Rendu-Osler-Weber disease or hereditary haemorrhagic telangiectasia. The PAVM may remain asymptomatic or manifest with hypoxemia, hemoptysis, hemothorax and/or neurologic symptoms. Despite its low incidence, the PAVM represent a serious entity that must be taken into account since untreated patients are in risk and present a poor prognosis. Transcatheter embolization with coils or detachable balloons is currently accepted as the treatment of choice. We present 6 patients (5 women and one man) who underwent embolotherapy of PAVM showing the long-term clinical and radiological follow-up. PMID- 19303486 TI - [Septal perforation in cocaine abusers: utility of the computed tomography]. AB - OBJECTIVE: Utility of computed tomography (CT) in the evaluation of the lesions of the sinonasal tract in chronic cocaine abusers. MATERIAL AND METHODS: Eleven young cocaine abusers were included consecutively. A CT of the paranasal sinuses was made by otorhinolaryngological symptoms. RESULTS: CT showed septal perforation in all of them. There was one nasal cavity and findings of sinusitis. Only a patient had naso-oral fistula. CONCLUSION: CT is useful in septal perforation in cocaine addicts because it delineates the extension of destructive lesions. PMID- 19303487 TI - [Case 1. Limbic encephalitis]. PMID- 19303488 TI - [Case 2. Congenital defect of the posterior arch of the atlas]. PMID- 19303489 TI - [Case 3. Congenital bronchial atresia]. PMID- 19303490 TI - [Case 4. Occipital sinus pericranii]. PMID- 19303493 TI - [Solution to case 3. Primary malignant melanoma of the rectum]. PMID- 19303494 TI - [Hysterosalpingography in a pregnant woman]. PMID- 19303495 TI - Venous thromboembolism: From bench to bedside. PMID- 19303496 TI - Effects of race and ethnicity on the incidence of venous thromboembolism. AB - Although there is strong evidence that the prevalence of venous thrombo-embolism (VTE) varies significantly among different ethnic/racial groups, the genetic, physiologic and/or clinical basis for these differences remain largely undefined. African-American patients have a significantly higher rate of incident VTE, particularly following exposure to a provoking risk factor such as surgery, medical illness, trauma, etc. In addition, African-Americans are more likely to be diagnosed with pulmonary embolism (PE) than deep-vein thrombosis (DVT) compared to Caucasian and other racial groups. On the other extreme, Asians/Pacific Islanders have a 70% lower prevalence of VTE and this is true for both idiopathic VTE and provoked, or 'secondary', VTE. Hispanics have a significantly lower prevalence of VTE compared to Caucasians, but higher than Asians/Pacific Islanders. The incidence of recurrent VTE varies depending on gender, type of thromboembolic event and race. Further research is needed in order to determine the fundamental differences between racial/ethnic groups that explain the observed differences in the prevalence of VTE. Race/ethnicity should be considered an important factor in the risk-stratification of patients with suspected VTE or patients at some risk for developing VTE. PMID- 19303497 TI - Cancer-associated thrombosis: risk factors, candidate biomarkers and a risk model. AB - Cancer and its treatments are well-recognized risk factors for venous thrombo embolism (VTE). Although solid tumors have historically been associated with VTE, more recent data suggest similar risk in patients with hematologic malignancies as well. The risk of VTE is not equal for all cancer patients or even in the same patient over time. In addition to well-known risk factors such as tumor site, stage, chemotherapy and comorbidities, candidate biomarkers have recently been identified including platelet and leukocyte counts, tissue factor and P-selectin. A validated risk model incorporating some of these risk factors and biomarkers has been shown to be predictive of VTE in cancer. VTE is associated with mortality, morbidity, potential delay in treatments for cancer and consumption of scarce health-care resources. Hence, reducing VTE with targeted prophylaxis could improve outcomes for cancer patients. PMID- 19303498 TI - Unanswered questions in venous thromboembolism. AB - We have made great strides in the diagnosis, treatment and prevention of venous thromboembolism (VTE). Despite these advances, however, questions remain. Perhaps the most important unmet need is the development and implementation of strategies to increase the uptake of guidelines for thromboprophylaxis. VTE is largely preventable with appropriate prophylaxis. New oral anticoagulants have the potential to further streamline VTE prevention and treatment. Although heparin, low molecular weight heparins or fondaparinux are frequently used for thromboprophylaxis in hospitalized medical or surgical patients, these agents are not ideal for out-of-hospital use. There is now good evidence that patients undergoing major orthopaedic surgery require extended thromboprophylaxis. Medical patients may also benefit from extended prophylaxis. The new oral anticoagulants will be more convenient than existing agents for this purpose and will help physicians and patients to adhere to optimal preventive strategies. VTE treatment also may be simplified with the new oral anticoagulants. With a rapid onset of action, these drugs may obviate the need for a parenteral anticoagulant for initial therapy. In addition, the new agents have the potential to streamline extended VTE therapy because, unlike vitamin K antagonists, they can be given in fixed doses without the need for coagulation monitoring. PMID- 19303499 TI - How to identify new genetic risk factors for VTE? AB - Venous thrombosis is a common and complex disease in which genetic risk factors play a major role. At present 6 strong and about 20 weak genetic risk factors are known. Family and twin studies indicate that additional genetic risk factors remain to be identified in order to explain the full extent of the known heritability. This short narrative review discusses several of the approaches which can be taken to identify new risk factors that predispose to venous thrombosis. PMID- 19303500 TI - Discovering novel risk factors for venous thrombosis: a candidate-gene approach. AB - The candidate-gene approach can be used to locate and identify genetic variations that are associated with a particular phenotype. This gene-centric approach assumes that there exists important genetic variation within genes that can influence health. Identifying known genes which are candidates for the phenotype of interest can be accomplished using existing knowledge about biology or using findings from genome-wide association studies. Genetic variation can be characterized locally by single nucleotide polymorphisms (SNPs) or insertion deletions, or it can be characterized more broadly in terms of haplotypes and diplotypes, which usually need to be inferred statistically. As an example, we present a candidate-gene approach to identify novel associations between variation in 24 clotting genes and the risk of incident venous thrombosis. PMID- 19303501 TI - Pathophysiology of venous thrombosis. AB - Using Virchow's triad as framework, it is clear that alterations in any of its components (blood composition, the vessel wall, and blood flow) can influence the propensity for the development of venous thromboembolism. Each can also influence the others in ways that enhance or reduce thrombotic propensity. While past work has concentrated on blood components that influence thrombogenicity, their influences generally are manifest only after the thrombotic process has begun, and it is initiation of thrombosis on the vessel wall that is the least well understood. This brief review attempts to link risk factors such as stasis and cancer to the mechanisms that initiate thrombosis. PMID- 19303502 TI - Role of selectins and fibrinolysis in VTE. AB - Venous thromboembolism (VTE) accounts for an estimated 900,000 cases of deep venous thrombosis (DVT) and pulmonary embolism (PE) yearly, resulting in approximately 300,000 deaths [1]. For the past 150 years, Virchow's triad has encompassed the elements of venous thrombogenesis, including stasis, changes in the vessel wall, and thrombogenic changes in the blood. However, in the early 1970s, through the work of Gwendylen Stewart, a relationship between thrombosis and inflammation was suggested. In this review, we will address the role of selectins and fibrinolysis in the process of venous thrombogenesis. PMID- 19303503 TI - The role of procoagulants and anticoagulants in the development of venous thromboembolism. AB - Procoagulant and anticoagulant reactions play an important role in the regulation of thrombin formation during secondary hemostasis. Three phases can be recognized in the kinetics of thrombin formation: an initiation phase, a propagation phase and a termination phase. Dysregulation of thrombin formation during each of these phases by (hereditary) changes in the plasma concentration of pro- and anticoagulants contributes to the development of venous thrombosis. Most important seems the defective down-regulation of the prothrombinase activity during the termination phase. Procoagulant and anticoagulant proteins have important roles in the regulation of fibrin formation during secondary hemostasis. Under normal physiological conditions there is a delicate balance between the procoagulant and anticoagulant reactions. After damage to the vessel wall sufficient fibrin is formed to arrest bleeding and allow repair of the lesion without obstructing blood circulation. Venous thrombosis can be considered as a hemostatic process getting out of control, where massive fibrin formation has resulted in the formation of an obstructive thrombus. Such thrombus formation is believed to be facilitated by changes in the vessel wall, blood flow and the composition of the blood. During the past 50 years substantial progress has been made in our understanding of the enzymatic reactions involved in the hemostatic process. At the same time information has been obtained on particular changes in the composition of the blood which contribute to the development of venous thrombosis. Most of these changes concern the procoagulant and anticoagulant systems. In this paper I will briefly discuss how fibrin formation is regulated by procoagulant and anticoagulant reactions and how certain changes in these pathways contribute to the development of venous thrombosis. PMID- 19303504 TI - Role of fibrinolysis in obesity and thrombosis. AB - Obesity is a common disorder and a known risk factor for thrombotic complications. Development of obesity is associated with extensive modifications in adipose tissue involving adipogenesis, angiogenesis and extracellular matrix proteolysis. The fibrinolytic (plasminogen/plasmin) system plays an important role in these processes. Studies using a nutritionally induced obesity model in transgenic mice support a role of the fibrinolytic system in adipogenesis and obesity. Studies using venous or arterial thrombosis models in obese mice, with impaired fibrinolytic activity, confirm a prothrombotic risk associated with obesity. PMID- 19303505 TI - Novel anticoagulants and the future of anticoagulation. AB - Since its discovery during the first half of the 20th century by biochemists at the University of Wisconsin, warfarin (along with other vitamin K antagonists) has remained the only oral anticoagulant available to patients at risk for thromboembolism. After nearly 6 decades in clinical practice, we have learned much about warfarin. Although it is highly effective for most patients, warfarin has a number of undesirable attributes: significant inter- and intra-patient variability in dose-response, a narrow therapeutic index, a slow pharmacodynamic response, and numerous interactions with both diet as well as other medications. The negative characteristics associated with warfarin have inspired many clinicians, patients, and researchers to wonder if a better alternative can be discovered. To that end, at least three novel anticoagulant compounds are in the late stages of development and several others are progressing through earlier phases of investigation. This review will summarize the latest clinical trial data pertinent to several newer antithrombotic agents and discuss recent developments that impact the safety and challenges associated with warfarin and other vitamin K antagonists (VKA). PMID- 19303506 TI - Thrombolytic therapy for deep vein thrombosis: potential application of plasmin. AB - Plasmin is the prototype of a distinct class of "direct-acting" fibrinolytic agents, with biochemical and physiological attributes that are favorable for catheter-delivered thrombolytic therapy. Our studies indicate that plasmin is superior to plasminogen activators for hemostatic safety and thrombolytic efficacy in experimental models, and that plasmin has potential to avoid the bleeding risk that accompanies therapy of deep vein thrombosis with currently used thrombolytic agents. PMID- 19303507 TI - Valvular dysfunction and venous obstruction in the post-thrombotic syndrome. AB - Medical thrombosis physicians have tended to view deep vein thrombosis (DVT) as essentially a problem of biology and hematology. This "field condition" concept holds that in susceptible individuals, DVT risk factors lead to a global hypercoagulable state which results in venous thrombosis. However, less attention has been paid to the key problems of venous anatomy and physiology that lead to the post-thrombotic syndrome (PTS) and its major health consequences. The primary objectives of this article are: (1) to review the macroscopic pathophysiology of PTS; (2) to discuss the roles of valvular reflux and venous obstruction in the development of PTS; and (3) to explore new endovascular PTS prevention and treatment strategies that feature the prevention or elimination of valvular reflux and late venous obstruction to improve DVT patient outcomes. PMID- 19303508 TI - Dermal pathology, cellular biology, and inflammation in chronic venous disease. AB - The pathophysiology of venous dermal pathology in chronic venous disease (CVD) is reflective of a complex interplay that involves sustained venous hypertension, inflammation, cytokine and matrix metalloproteinase (MMP) activation, and altered cellular function. Endothelial expression of specific adhesion molecules recruits leukocytes, and diapedesis of these cells into the dermal microvasculature promotes an inflammatory response with activation of cytokines and proteinases. Altered cell function enhances a state of vulnerability in the surrounding tissues initiating specific changes associated with venous disease. Ultimately, the persistent inflammatory-proteinase activity leads to advanced chronic venous insufficiency (CVI) and ulcer formation. PMID- 19303509 TI - Thrombus resolution and vein wall injury: dependence on chemokines and leukocytes. AB - Chronic venous insufficiency resulting in post-thrombotic syndrome occurs commonly after acute deep vein thrombosis, and is a prevalent cause of vascular disease morbidity in the community. Therefore, a better understanding of the pathophysiologic mechanisms that promote the development of chronic venous insufficiency could lead to novel approaches to interrupt the natural history and prevent post-thrombotic syndrome. In this paper, we will review the evidence that venous thrombus resolution is an inflammatory process that is dependent on chemokines and leukocytes. PMID- 19303510 TI - Images in emergency medicine. Guttate psoriasis. PMID- 19303511 TI - Images in emergency medicine. Open joint injury. PMID- 19303512 TI - Commentary: advocating for safety--the motorcycle helmet debate. PMID- 19303513 TI - The golden stethoscope. PMID- 19303514 TI - Annals of Emergency Medicine Journal Club. Empiric antibiotic therapy for sepsis patients: monotherapy with beta-lactam or beta-lactam plus an aminoglycoside? Answers to the November 2008 Journal Club questions. PMID- 19303515 TI - Images in emergency medicine. Traumatic tension pneumocephalus. PMID- 19303516 TI - Images in emergency medicine. Hair artifact. PMID- 19303517 TI - Levamisole found in patients using cocaine. PMID- 19303518 TI - The Sacrifice Festival: who is the victim? PMID- 19303519 TI - In response to "The unexpected challenges of accurate medication reconciliation". PMID- 19303520 TI - New form of tendinopathy discovered at Scientific Assembly. PMID- 19303521 TI - Emergency ultrasound guidelines. PMID- 19303522 TI - [SEPAR: work published in 2008]. PMID- 19303523 TI - [Innovations in the epidemiology, natural history, diagnosis and treatment of sleep apnea-hypopnea syndrome]. AB - The following review summarises the most important articles published on sleep apnea-hypopnea syndrome (SAHS) during the current year. The analysis of the many factors implicated in the risk of cardiovascular diseases associated with SAHS is currently of great interest to the scientific community. There are many studies on this subject that demonstrate the role of inflammatory and immunological mediators, their relationship with endothelial damage and their influence in the genesis of cardiovascular disease in patients with SAHS. The role of CPAP in preventing this cardiovascular risk has had varied results. Although there is no evidence of benefit or harm in its use in heart failure, in cerebrovascular accidents SAHS has been confirmed as a predisposing factor and the reported increase in mortality would justify the intention to treat SAHS in these patients. Likewise, the reduction in blood pressure found with CPAP treatment could reduce the risk of cardio-cerebrovascular disease. The recent knowledge that there is expression of multiple phenotypes of SAHS gives a glimpse in the future of a disease based on different specific phenotypes, where the traditional symptomatology that defined the syndrome does not limit its treatment. To obtain a reliable and cost-effective diagnostic method that responds to the demands of the public health problem that is SAHS, particularly in sectors of the population that remain under-diagnosed and less well known, such as children, women and the elderly population is another one of the challenges reflected in published studies. In short, the growing knowledge on the biology of SAHS, its cardiovascular implications and its effect on the morbidity and mortality of the population will enable us to understand the true dimension of this disease in the next few years. PMID- 19303524 TI - [Respiratory infections research: a perspective from the tuberculosis and respiratory infections area (TIR)]. AB - The scientific production of the TIR Area of SEPAR during 2008 is reviewed. In pneumonias, studies on C-reactive protein, procalcitonin and the cytokines as predictive markers of treatment failure are noteworthy, as well as research into the genetic predisposition of the host (polymorphisms of mannose binding lectin) in the prognosis. Among the different activities on tuberculosis in the SEPAR <> year, was the publication of the new SEPAR guidelines for the <>. The studies into tuberculosis have been on, the tuberculosis infection, the new in vitro techniques for detecting interferon gamma, new non-bacillary tuberculosis diagnostic committees, and treatment schemes without rifampicin and isoniazid. In COPD,we have highlighted new aspects in the indications for antibiotic treatment in the Consensus Document for the antibiotic treatment of acute exacerbations of COPD, and in the SEPAR-ALAT Clinical Guidelines. In the field of cystic fibrosis (CF), we highlight 3 studies: a) association between colonising- Pseudomonas aeruginosa induced chronic infection and bronchial hyperreactivity; b) serum immunoglobulins response to Aspergillus fumigatus and Candida albicans in the colonising of the lower respiratory tract and its clinical significance; and c) prevalence of environmental mycobacteria in these patients. In the chapter on bronchiectasis, a study on the relationship between systemic inflammation and severity parameters is highlighted, and finally, the main contributions of the new SEPAR guidelines on the diagnosis and treatment of bronchiectasis. PMID- 19303525 TI - [Present and future of the SEPAR Smoking and Health Area]. AB - Smoking is the primary avoidable cause of premature death and morbidity in our country. According to data from the WHO, tobacco costs more than 5 million lives per year, a figure that is assumed to continue increasing and will be 10 million by the year 2030. As health professionals we are obliged to be actively involved in this serious health problem, by correctly identifying, diagnosing and treating all smokers to help them in the process of giving up and help them achieve total abstinence. There is scientific evidence that smoking is the direct cause and reason for the worsening of many respiratory diseases, which due to their nature are basically the responsibility of the pneumologist, who officially, according to the Health Service Administration, is the specialist and expert in this area. The Smoking and Health Area of SEPAR was started in 1995. Its primary objectives were to increase the awareness of the smoking problem among the members of our Scientific Society, improve their scientific knowledge of this diseases, to help in diagnostic and treatment interventions in smokers, and lastly, to promote interventions for the prevention and treatment of smoking by the health and political Administrations. Nowadays it is one of the most dynamic areas of SEPAR and has more than 400 members. There are many unresolved challenges in our area, but the fundamental one which continues to occupy a particular place for the Administration, for our patients, for other health professionals of other specialties and scientific societies, and for all SEPAR professionals, is a key aspect of which there is no doubt: to defend the health and well-being of our population against the main cause of avoidable disease and death, tobacco. PMID- 19303526 TI - [Update in respiratory disease and environmental exposure: an invisible relationship]. AB - Significant contributions have been made in the past year on different aspects of occupational/environmental respiratory disease. In the case of neoplastic diseases associated with asbestos inhalation, the areas of most interest have been in the search for tumour markers, the importance of the determination of asbestos fibre deposits in biological samples, and new therapeutic schemes in malignant pleural mesothelioma. A consensus article has been published on occupational asthma, in which some clinical evidenced-based recommendations are established, directed at the diagnosis and management of work-related asthma. As regards hypersensitivity-induced pneumonitis, the clinical and evolutionary aspects of this disease have been described in a large series of 86 patients with pigeon-fancier lung. There have also been interesting studies published this year that emphasise the need to take an occupational history in patients with respiratory symptoms in order to look for a causal or synergic relationship with smoking. Finally, the results of studies have been published which were directed at elucidating the role of urban contamination, mainly caused by road traffic, in the deterioration of lung function. A recent study showed that it would be possible to achieve a significant reduction in urban mortality attributed to urban contamination by reducing the levels of PM 2.5. They conclude that more restrictive standards need to be adopted in Europe to protect the health of the population, which coincides with the proposal by the World Health Organisation. PMID- 19303527 TI - [Asthma clinical practice guidelines: advantages and pitfalls]. AB - The Clinical Practice Guidelines on asthma have contributed towards unifying concepts and reaching a consensus on performances between different professional groups. However, they have failed in the overall improvement in the management of asthma, the final objective that they are meant to achieve. Today, almost 20 years after they appeared, the majority of asthmatic patients are still inadequately controlled, partly due to lack of follow up by doctors and the rest of health care staff who have to look after them. This lack of follow up of these recommendations is probably associated with a lack of well structured planning in their circulation and implementation. Also, although the recommendations of these guidelines agree in what is essential, they differ in other aspects, which in turn could be determining factors in clinical practice. The purpose of this article has been to establish the main differences in the recommendations that the principal clinical practice guidelines on the disease propose on the diagnosis, classification and treatment of asthma. To do this we have compared, The British Guideline on the Management of Asthma 2007, The Global Strategy for Asthma Management and Prevention/Global Initiative for Asthma 2006 (GINA), the National Prevention program for Education on Asthma (Programa Nacional de Prevencion para la Educacion del Asma) (NAEPP), the Spanish Guide for the Management of Asthma (Guia Espanola para el Manejo del Asma 2003) (GEMA) and the ALAT y SEPAR guides, Latin-America and Spain. Recommendations for the Prevention and Treatment of Asthma Exacerbation (America Latina y Espana. Recomendaciones para la Prevencion y el Tratamiento de la Exacerbacion Asmatica 2008) (ALERTA). PMID- 19303528 TI - [Annual Review of COPD]. AB - There have been significant advances in the knowledge in the thoracic and extrathoracic aspects of chronic obstructive pulmonary disease (COPD) in the past few years. COPD is associated with numerous comorbidities, the prevalences of which have recently been evaluated. Dyspnea has been shown to be associated with neuropsychiatric disturbances, such as anxiety. Muscular dysfunction has been associated with inflammation and oxidative stress, in which respiratory muscle satellite cells play an important role in repair. Respiratory rehabilitation and physiotherapy must form an important part of individualised patient treatment analogous to the pharmacological treatment. As regards acute exacerbations, infection is the cause of 75% of them, sputum characteristics and the suspicion of Pseudomonad being key factors in the antibiotic treatment. Questions, such as markers which can detect the origin of the infection, prognostic factors, or the role of short stay pneumology units, are of particular importance. The variability in COPD treatments and the lack of suitable international clinical guidelines, continue to be subjects of debate. To the poor use of the treatment schemes in the guidelines, can be added the irregular uses of inhaled medication, the insufficient use of medical advice or the low intervention in cigarette smoking in all age groups. PMID- 19303529 TI - [Thoracic oncology in Archivos de Bronconeumologia 2008]. AB - The articles on thoracic oncology published in this journal during the year 2008 are briefly commented on. As regards the epidemiology, it is noted that the standardised incidence rate of lung cancer in males is starting to decline, and there is another study that analysed links between two common diseases, COPD and lung cancer. Other works have focused on aspects such as diagnosis, staging or prognosis, and analysing the value of positron emission tomography in the assessment of a solitary pulmonary nodule, the effectiveness of aspiration transbronchial needle aspiration in mediastinal staging, the prognostic significance of the over-expression and amplification of c-erbB-2 in patients with small cell carcinoma. As regards treatment, other authors analysed the survival of patients with N2 lung cancer detected during or after lung resection surgery. The new therapeutic technique of ablation of lung tumours by radiofrequency in the early stages is the subject of two publications that describe its basis, indications, contraindications and first results. Lastly, the communication skills needed to inform patients, surgery of lung metastases, and the presentation of an unusual case of carcinoid tumour were the subjects of other articles. PMID- 19303530 TI - [Annual review of Archivos de Neumologia in interventional pneumology, interstitial diseases and lung transplantation]. AB - A review has been made of original articles on invasive pneumology techniques, interstitial diseases and lung transplantation, published in the Archivos de Bronconeumologia during the year 2008. We have selected the publication by Martinez-Olondrins et al on the mediastinal staging of bronchogenic carcinoma by <> transbronchial needle aspiration to highlight the role of this simple, safe and cost-effective technique at a time when aspiration by ultrasound-guided bronchoscopy is profiled as an alternative to staging by mediastinoscopy. Besides its usefulness in the study of lymph nodes, transbronchial needle aspiration increases the overall performance of bronchoscopy by 20%, which means that it should be considered as a basic tool in the study of lung cancer. We also comment on the work by Galvis-Caravajal et al, who describe percutaneous radiofrequency as an alternative to radiotherapy in small lung or metastasic tumours. In diffuse interstitial disease, Morell et al analysed the diagnostic methods in 500 patients with this clinical-radiological presentation in which a definitive diagnosis was achieved in 85%, with 25% of them being obtained by non-invasively. Baloira et al analysed the characteristics of 19 patients with desquamative interstitial pneumonia and respiratory bronchiolitis associated-interstitial lung disease obtained from the National register of Interstitial Diseases. PMID- 19303531 TI - [Pulmonary arterial hypertension: a voyage around the year 2008]. AB - There have been spectacular developments in pulmonary arterial hypertension (PAH), both in its treatment and knowledge of its pathogenesis. Several studies have been published throughout 2008 that have contributed to improve these two aspects a little. As regards the pathogenesis, mutations in BMPR2 continue gaining points as fundamental factors in the development of the disease. It has been shown that patients who carry any of them have a more rapid and severe clinical course. There is a relationship between the BMPR2 pathway and inflammation of the pulmonary vascular tree. A new anti-endothelin drug, ambrisentan, has also appeared on the scene this year. With an efficacy comparable to other drugs of its group, the secondary effects appear to be a lot less. An important work has been the demonstration of an improvement in several parameters in functional class II in patients with PAH with bosentan. Results using new combinations, such as sildenafil and epoprostenol, have also been presented. A common type of PAH is that which seems to be associated with thromboembolic disease. Treatment with sildenafil and in some selected cases, percutaneous angioplasty, has obtained favourable responses. Finally, in 2008, two new consensus documents have emerged, one Spanish and the other British, which in the light of current knowledge, give a clearer insight into the management of this serious disease. PMID- 19303532 TI - [Horizons in respiratory care]. AB - As nurses and physiotherapists, we truly wish that our work could be rewarded with sufficient time, support, training and recognition so that it could be reflected in studies, publications and research. It does not seem right and it is unjust that the care profession framed within a multidisciplinary need and reality, with the largest number of professionals and hours of health care, does not have the recognition of its space. Too much blame. We need that space to continue growing and in particular to improve the quality of life or our population. Stop looking back and consolidate ourselves in the new possibilities that are being proposed to us without being dragged down by the autocracy of the market. Perhaps it is time to review and give each thing its label. Lit is our situation and to many of us it does not motivate us, or allow us to flow, but something is changing and change is the only thing that is permanent. PMID- 19303533 TI - [Irritable bowel syndrome: current aspects]. PMID- 19303534 TI - [Somatic comorbidities in irritable bowel syndrome: fibromyalgia, chronic fatigue syndrome, and interstitial cystitis]. AB - Fibromyalgia, chronic fatigue syndrome, and interstitial cystitis frequently overlap with irritable bowel syndrome (IBS). There is a positive correlation between the incidence of these comorbidities and increased health care seeking, reduction in quality of life, and higher levels of mood disorders, which raises the question of a common underlying pathophysiology. A possible central hypersensitization disorder seems to be particularly involved in the dysfunction of bidirectional neural pathways and viscerovisceral cross-interactions within the CNS, thus explaining these many extraintestinal manifestations in IBS. PMID- 19303535 TI - [How can an effective drug to treat irritable bowel syndrome be successfully developed?]. AB - BACKGROUND: Irritable bowel syndrome (IBS) is a prevalent disorder, and although the pharmaceutical industry knows the potential fallout of a successful drug launch in this area, effective drug treatments are rare. AIM: To give an overview of the main factors interfering with the development of IBS drugs and to provide pertinent methodological indications to improve their investigation in clinical trials. RESULTS: Developing IBS drugs remains a major challenge, as numerous factors, related or unrelated to the nature of the disease itself, interfere with the demonstration of efficacy : the multiplicity of physiopathological mechanisms, wide variation in symptoms across patients and over time, associated psychological traits and environmental aspects, and a very significant placebo effect. There can be no question of developing drugs to target a single receptor in the hope of thereby impacting the whole range of factors involved in the genesis of IBS symptoms. Drug safety is, moreover, a prime consideration, given that this pathology, while certainly disabling, is not life-threatening. If a significant difference between a new treatment and placebo is to be demonstrated on a clinical trial, inclusion and efficacy criteria and study treatment duration must be predefined very precisely. The primary endpoint is abdominal pain, but the assessment of relief of the patient's symptoms has been also recommended, even if there is as yet no consensus as to its definition. The impact of a new IBS drug on patient's quality of life is an important secondary endpoint. CONCLUSION: In IBS more, perhaps, than in other pathologies, study design needs very careful consideration if new IBS drug trials are to be conclusive. However, some critical methodological issues (e.g., definite primary endpoint, interpretation of results, and definition of responders) are still unresolved. PMID- 19303536 TI - [Irritable bowel syndrome: epidemiology/economic burden]. AB - The prevalence of irritable bowel syndrome (IBS) is high wherever it is assessed. The definition is still controversial, but the IBS patient's characteristics are quite similar in industrialized countries. The lack of a definite biological marker results in the prescription of many complementary examinations. The diagnosis of IBS remains an elimination diagnosis, despite the publication of the Rome criteria, which has attempted to advance toward a positive diagnosis. Abdominal pain or discomfort are the main symptoms and are the major reasons for consulting. The quality of life is impaired in IBS and is clearly correlated with symptom severity. All these points at least partially explain the economic burden of IBS on health care resources. The cost of IBS is difficult to assess because of its different components: the direct costs such as the number of medical consultations, drug consumption, and the number and quality of complementary investigations are easier to evaluate than the indirect costs such as work absenteeism or altered presenteeism, which can have a major impact on the cost of IBS. PMID- 19303537 TI - [Intestinal motility disorders and visceral hypersensitivity in irritable bowel syndrome]. AB - The pathophysiology of irritable bowel syndrome is complex, secondary to a dysregulation of the visceral neuromuscular system. Motor disorders and digestive hypersensitivity have been studied most: phasic and tonic abnormalities and visceral hypersensitivity associated with different types of receptors have been identified in patients with irritable bowel syndrome. For other patients, the combination of these disorders can explain abnormal physiological responses. Some records of various explorations are correlated with patient symptoms, such as pain, bloating, or bowel movement disorders. These sensorimotor disorders, probably not limited to the gut, are reviewed in this article. PMID- 19303538 TI - [Alterations of intestinal epithelial barrier and flora in the irritable bowel syndrome]. AB - The irritable bowel syndrome (IBS) is a common bowel disorder that markedly impairs quality of life of patients. The causes of IBS symptoms are not well known. Motility and sensibility disorders, a genetic susceptibility, stress, previous GI infections and food intolerance have been involved in the pathogenesis of IBS. Recent investigations suggest that alterations of the intestinal epithelial barrier integrity, in particular increased permeability, could modify the cross-talk between bacterial microflora and the host, thus leading to persitent "low-grade" inflammation and alterated GI motility and sensitivity. PMID- 19303539 TI - [Brain-gut axis dysfunction]. AB - There is a bidirectional relation between the central nervous system and the digestive tract, i.e., the brain-gut axis. Numerous data argue for a dysfunction of the brain-gut axis in the pathophysiology of irritable bowel syndrome (IBS). Visceral hypersensitivity is a marker of IBS as well as of an abnormality of the brain-gut axis. This visceral hypersensitivity is peripheral and/or central in origin and may be the consequence of digestive inflammation or an anomaly of the nociceptive message treatment at the spinal and/or supraspinal level. Stress is involved in the genesis and maintenance of IBS. Disturbances of the autonomic nervous system are observed in IBS as a consequence of brain-gut axis dysfunction. The contribution of the neurosciences, in particular brain imaging techniques, has contributed to the better understanding of IBS physiopathology. The better knowledge of brain-gut axis dysfunction has therapeutic implications, either through drugs and/or cognitive and behavioral therapies. PMID- 19303540 TI - [Therapeutic targets]. AB - Based on better recent knowledge of the factors involved in triggering visceral hyperalgesia, the therapeutic approach to irritable bowel syndrome (IBS) treatment is changing. The classical approach targeting first bowel movement alterations or motility disorders using spasmolytic agents has to be replaced by visceral antinociceptive drugs. Several mediators and receptors involved in gut hyperalgesia have already been identified. Serotonin (5-HT), tachykinins, CCK, NGF, and other mediators are involved in experimental models of gut hyperalgesia, and related receptor antagonists have already been introduced in clinical trials. However, IBS is associated with mucosal immune stimulation, considered a microinflammatory state associated with increased density of immunocytes and mast cells, offering new targets. Altered mucosal barrier permeability with increased entry of toxins and bacteria is considered to be responsible for the mucosal microinflammation. Endogenous but predominantly luminal factors have been identified as factors responsible for such altered permeability. These clinical data have opened the door to promising future drugs able to prevent or blunt such permeability alteration, which therefore may constitute a pathophysiological treatment for IBS. PMID- 19303541 TI - [Irritable bowel syndrome: dietary and pharmacological therapeutic options]. AB - In irritable bowel syndrome, the main objectives of the treatment are the relief of abdominal pain then the improvement of bowel disturbances. Spasmolytic agents, or clays remain routinely the first line pharmacological options. The efficacy of dietary recommendations is not validated in most of the cases while dietary fibers, mainly insoluble fibers, may even worsen abdominal discomfort. In C-IBS, osmotic laxatives or macrogol are effective to improve colonic transit while loperamide and also colestyramine can be prescribed to reduce the number of stools of D-IBS patients. When the first line treatment fails to improve symptoms, antidepressants (tricyclic rather than SSRs) can be prescribed at lower doses than that recommended for depression. In meta-analysis, the odds ratio for pain relief varies from 2 to 4 and strongly depends on the patient's compliance to the treatment. Probiotics, pregabalin and even antibiotics (i.e neomycin, metronidazole or rifaximin), are possible new therapeutic options. Few clinical trials suggest that ramosetron (a new 5HT3 antagonist), octreotide, melatonin, or lidocain could be also discussed in the future. A non pharmacological therapeutic approach has to be considered, particularly in patients with severe symptoms, in combination with pharmacological treatment. PMID- 19303542 TI - [Alternative treatments in irritable bowel syndrome]. AB - Managing irritable bowel syndrome (IBS) is difficult and often a source of dissatisfaction for the patient, explaining the increasingly frequent recourse to alternative treatments. These highly varied treatments are often associated. They can be classed into four categories: reflexology methods, interventions on the psyche, biological therapies, and treatments using certain forms of energy. Although some studies show interesting results, currently there are not sufficient scientific arguments to recommend one or another of these alternative treatments. Multicenter controlled studies are needed to better evaluate the strategies that appear to be cost-effective. PMID- 19303543 TI - [Irritable bowel syndrome: outcome aspects]. AB - Irritable bowel syndrome is a chronic relapsing disorder. In most patients, symptom severity and clinical characteristics change frequently. Nonetheless, broad progressive trends can be defined. Over the very short term, symptoms occur over short sequences appearing sequentially and are interrupted by short remission periods. Over the medium term (3 months), early recovery from symptoms is observed in 20%-40% of patients. No good clinical predictors of IBS progression seem to exist even if clinical improvement at 3 months seems related to a better prognosis after a 1-year follow-up. Over the long term (several years), symptoms are still present in 60%-70% of patients. Symptom turnover is high in these patients from either transitioning into other irritable bowel syndrome subgroups or overlapping with dyspeptic symptoms. Psychiatric comorbidities appear to be of primary importance in the clinical course of patients with irritable bowel syndrome. PMID- 19303544 TI - [Diagnosis of irritable bowel syndrome]. AB - The diagnosis of irritable bowel syndrome (IBS) is a clinical diagnosis, based on the Rome III criteria. The identification of alarm features (red flags), associated with an organic digestive pathology is a key point during the initial screening. Age over 50 years, blood in feces, and diarrhea seem to be the three most specific red flags. Biological and morphological examinations must be performed cautiously; their profitability is low in patients with Rome criteria. In the majority of clinical situations encountered in daily clinical practice, a positive diagnosis of IBS can be made. PMID- 19303546 TI - Preface. Nutritional topics update. PMID- 19303547 TI - Clinical assessment of nutritional status and feeding programs in horses. AB - Veterinarians are a primary source of nutritional information and advice for horse owners. This article reviews methods for clinical assessment of nutritional status and feeding programs that can be applied to an individual horse or group of horses. Physical examination, including measurement of body weight and evaluation of body condition score, estimation of nutrient requirements and the nutrient content of the horse's diet, and evaluation of the feeding method are important components of the assessment. Ongoing clinical assessment of health and body condition will gauge the need for reassessment of the feeding plan. Obvious indications for prompt reevaluation of diet and feeding include changes in health status (eg, body condition), life stage or physiologic state (eg, pregnancy), or performance status. PMID- 19303548 TI - Primer on dietary carbohydrates and utility of the glycemic index in equine nutrition. AB - The feeding of "low carbohydrate" or "low glycemic" diets has been recommended for management of horses with metabolic and endocrine disorders in which insulin resistance is a component. A "low carbohydrate" diet is a misnomer, however, because horses require that a significant proportion of their daily ration comprises structural carbohydrates (fiber/forage) to maintain gut health and mental wellbeing. This article provides a detailed description of the different carbohydrates in equine feeds. It also reviews the terminology used to describe glucose and insulin responses to the ingestion of carbohydrates, in particular the concept of the glycemic index. Some of the factors that influence glycemic index in humans and the glycemic response to a meal in horses are also discussed. PMID- 19303549 TI - Pasture-associated laminitis. AB - Laminitis is a painful and debilitating condition of horses and ponies that has major economic and welfare implications. Anecdotal observations and the results of survey studies have indicated that most laminitis cases occur in horses and ponies kept at pasture (hence, the term pasture-associated laminitis). Risk for development of pasture-associated laminitis represents a dynamic interaction between animal predisposing factors (an insulin-resistant phenotype commonly termed equine metabolic syndrome) and environmental conditions, particularly the nonstructural carbohydrate (simple sugars, starches, and fructans) content of pasture forage. Countermeasures for avoidance of pasture-associated laminitis involve (1) mitigation of metabolic predisposition (insulin resistance and obesity) in high-risk horses and ponies and (2) dietary and pasture grazing management strategies that minimize exposure to the dietary conditions known to trigger laminitis in susceptible animals. PMID- 19303550 TI - Dietary management of obesity and insulin resistance: countering risk for laminitis. AB - Insulin resistance (IR) and hyperinsulinemia increase risk for development of laminitis in horses and ponies. Obesity also has been associated with heightened risk for laminitis, likely by means of development of IR. Dietary factors, particularly the nonstructural carbohydrate (NSC) load, modulate risk for laminitis in these animals by means of exacerbation of IR or gastrointestinal disturbances that trigger the condition. Specific dietary management strategies to lessen risk for laminitis include caloric restriction to promote weight loss and improve insulin sensitivity in obese animals and strict control of dietary NSCs, with elimination of grains and sweet feeds from the ration and restricted access to pastures that may be rich in NSCs. Medical treatment with levothyroxine or metformin may be indicated in animals that do not respond to conservative dietary management. PMID- 19303551 TI - The role of nutrition in colic. AB - Nutritional intolerances manifesting as colic in the horse may be largely explained by divergence from the diet and ingestive behaviors to which the feral ancestors of modern domesticated equids had become accustomed and adapted. High starch diets and abrupt dietary changes are probably foremost in the risk factors for diet-associated colic in the horse and have their basis in disruption of the stability of microbial populations resident within the equine hindgut. Although some general associations between colic and diet may be inferred from several epidemiologic studies, data derived from studies of single and specific disease processes associated with colic allow more effective practical application of corrective dietary management strategies in situations where colic risk is judged to be increased. PMID- 19303552 TI - Nutrition and dietary management of equine gastric ulcer syndrome. AB - Equine gastric ulcer syndrome (EGUS) is common in horses. Diagnosis is based on history, clinical signs, gastroscopic examination, and response to treatment. Effective pharmacologic agents are available to treat EGUS, but more comprehensive measures of environmental and dietary management are needed to decrease ulcer severity and recurrence. This article provides an understanding of dietary components and how feeds interact with stomach mucosal barrier function to cause EGUS. In addition, a secondary goal is to provide information on how diet and environmental management can reduce ulcer severity and prevent recurrence in horses with EGUS. PMID- 19303553 TI - Nutrition of critically ill horses. AB - Nutritional supplementation is becoming the standard of practice in equine medicine, although there are minimal data on nutritional support in critically ill horses and its association or effect on morbidity and mortality or length of hospital stay. Horses can be fed orally and when that is not possible, intravenously or parenterally. Enteral feeding is less expensive, more physiologic, improves immunity, and is easier and safer. This article reviews available information on the development of a nutritional plan for critically ill horses, and describes methods for and complications of enteral and parenteral feeding. PMID- 19303554 TI - Feeding management of sick neonatal foals. AB - Nutritional support of the foal can be challenging because of the constant changes in nutritional requirements and dietary composition during the transition from neonate to weanling. Additional complexity arises because of dilemmas regarding the means and route of delivery of nutrition to the foal, and the possibility that metabolic dysfunction may impair the ability of the foal to use nutrients appropriately. This article provides practical information on enteral and parenteral nutritional support of sick neonatal foals. The potential benefits of a conservative, hypocaloric feeding strategy, particularly in the very sick patient, are also discussed. PMID- 19303555 TI - Optimal diet of horses with chronic exertional myopathies. AB - Chronic exertional rhabdomyolysis represents a syndrome of recurrent exercise associated muscle damage in horses that arises from a variety of etiologies. Major advances have been made in the understanding of the pathophysiology of this disease, and causative genetic defects have been recently identified for two conditions-polysaccharide storage myopathy of quarter horses, paints, warm bloods, and draft breeds. Dietary management in combination with a regular exercise regimen comprises the most effective means for control of clinical signs. PMID- 19303556 TI - Feeding management of elite endurance horses. AB - This article reviews the principles of feeding management for endurance horses. The amount and type of dietary energy (calories) are key considerations in dietary management, because (1) there is evidence that the body condition score, an indicator of overall energy balance, influences endurance exercise performance, and (2) the source of dietary energy (ie, carbohydrate versus fat calories) impacts health, metabolism, and athletic performance. Optimal performance is also dependent on provision of adequate feed, water, and electrolytes on race day. PMID- 19303557 TI - Nutrition of the aged horse. AB - This article reviews current thoughts on nutrition of the older horse in health and disease. Common causes of weight loss and poor body condition in old horses include dental or oral cavity abnormalities, pituitary pars intermedia dysfunction, and reduced feed intake attributable to competition from herd mates or pain associated with osteoarthritis. Feed intake and body condition may improve after institution of management changes. Thin but otherwise healthy old horses can benefit from a diet that provides 12% to 16% crude protein and includes highly digestible feedstuffs. In horses with severe irreversible dental problems, long-stem fiber (hay) should be replaced by soaked hay cubes, short chopped hay, or heavily soaked sugar beet pulp. Evidence of chronic endocrine, hepatic, or renal disease dictates dietary modifications. PMID- 19303558 TI - Role of diet and feeding in normal and stereotypic behaviors in horses. AB - This article reviews the effects of diet on equine feeding behavior and feeding patterns, before considering the evidence that diet affects reactivity in horses. A growing body of work suggests that fat- and fiber-based diets may result in calmer patterns of behavior, and possible mechanisms that may underpin these effects are discussed. In contrast, there is little evidence that herbal- or tryptophan-containing supplements influence equine behavior in any measurable way. The role of diet in the development of abnormal oral behaviors, particularly the oral stereotypy crib-biting, is also reviewed, and suggestions for future work are presented. PMID- 19303559 TI - Comment on "Carotid chemodectomas: long-term results of subadventitial resection with deliberate external carotid resection". PMID- 19303560 TI - Using common iliac artery as an inflow for creation of lower extremity vascular access: a new approach. PMID- 19303561 TI - New views on Clostridium difficile infections: Introduction. PMID- 19303562 TI - Characterisation of Clostridium difficile isolates by slpA and tcdC gene sequencing. AB - The genotyping of Clostridium difficile is generally performed by the analysis of fragment- or amplification-length polymorphism by pulsed field gel electrophoresis (PFGE) or polymerase chain reaction (PCR) ribotyping. However, sequence-based methods allow typing technique standardisation and data comparison. In the present study 100 C. difficile isolates, obtained from various institutions in the state of Saarland, Germany, were prospectively analyzed by surface layer protein A single locus sequence typing (slpAST). A high proportion (52%) of isolates attributable to ribotype 027 (RT027) was found indicating that the new outbreak strain has become endemic, at least in parts of Germany. RT027 strains displayed characteristic mutations of the potential toxin repressor gene tcdC and antibiotic resistance to macrolides and fluoroquinolones. C. difficile isolates attributable to ribotypes RT001 (27%), RT014/066 (5%), RT078 (4%), to the smz genotype (3%), and to more sporadic genotypes were also identified. Overall, the prevalence of strains with resistance to macrolides or fluoroquinolones was >80%. slpAST allows the comprehensive identification of C. difficile strains by global data comparison, exemplified here by our identification of smz strains previously identified by slpAST of a Japanese outbreak. In conclusion, slpAST appears to be a powerful discriminative tool for the straightforward, standardised genotyping of C. difficile isolates. PMID- 19303563 TI - Surveillance of Clostridium difficile-associated diarrhoea with the German nosocomial infection surveillance system KISS (CDAD-KISS). AB - A study of 2856 Clostridium difficile -associated diarrhoea (CDAD) patients from 34 hospitals in Germany was carried out over a period of 12 months in 2007. The overall incidence of CDAD cases was 46.5 per 10,000 admitted patients, or 6.6 cases per 10,000 patient-days. Seventy-three percent of cases were considered to be nosocomial and 8.4% were classified as severe. There was a wide range in the incidence of CDAD between the hospitals (interquartile range [IQR] 3.2-9.2 cases per 10,000 patient-days), and the incidence of nosocomial cases (IQR 1.7-6.7 per 10,000 patient-days). The differences between the hospitals were in line with the differences in local diagnostic and infection control procedures. The overall incidence in the hospitals participating in the surveillance system was much higher than that in the discharge data from the whole of Germany in 2006. This may be due to more diligent ICD9 coding by the clinicians during an active surveillance strategy. However, the group of hospitals participating in the study was small and may not be representative of the situation in the country as a whole. PMID- 19303564 TI - Clostridium difficile 30 years on: what has, or has not, changed and why? AB - The report of clindamycin-associated colitis in 1974 by Tedesco et al. [Ann Intern Med 81: 429-33] stimulated an intense search for the cause of this severe complication of antibiotic use. The search culminated in early 1978 in the publication of a series of papers within 3 months that identified the causative agent as Clostridium difficile and its accompanying toxins. Thirty years later we are in the midst of a resurgence of C. difficile infection (CDI) in North America and Europe that is greater than ever previously reported and for which morbidity and mortality appear to be higher than ever seen in the past. The purpose of this review is to highlight the discoveries of the past 30 years that, in my view, have brought us to our current level of understanding of the pathogenesis, prevention and treatment of CDI, and to suggest why a disease thought to be managed so well 30 years ago could now be causing more morbidity and mortality than ever before. In the 21st century the focus should be on better understanding the relationship between the C. difficile organism and the host at the mucosal level, so that biotherapeutic and vaccine strategies for the prevention of CDI can be developed. PMID- 19303565 TI - New trends in Clostridium difficile virulence and pathogenesis. AB - The disease spectrum caused by Clostridium difficile infection ranges from antibiotic-associated diarrhoea to life-threatening clinical manifestations such as pseudomembranous colitis. C. difficile infection is precipitated by antimicrobial therapy that causes a disruption of the normal colonic microbiota, predisposing to C. difficile intestinal colonisation. The pathogenicity of C. difficile is mediated by two exotoxins, TcdA and TcdB, both of which damage the human colonic mucosa and are potent cytotoxic enzymes. C. difficile must first be implanted in the gut and attach to epithelial cells, which are protected by a layer of dense mucus. Confirmed and putative accessory virulence factors that could play a role in adherence and intestinal colonisation have been identified and include proteolytic enzymes and adhesins. Recently, the epidemiology of C. difficile infection has radically changed and an increased incidence is associated with outbreaks in North America and Europe. Several reports suggest that disease severity is increasing to include sepsis syndrome and toxin megacolon. Elderly, debilitated patients in hospitals and nursing homes are particularly vulnerable. A hypervirulent, epidemic strain has been associated with the changing epidemiology and severity of disease. Here, we review the characteristics of the epidemic NAP1, PCR ribotype 027 C. difficile strain that could explain its hypervirulence and epidemic spread. PMID- 19303566 TI - Clostridium difficile and fluoroquinolones: is there a link? AB - Clostridium difficile infection has become one of the most significant threats to hospitalised patients and represents an increasingly important issue in terms of morbidity and mortality within our institutions. The arrival of a new, more virulent strain coincided with the introduction of several new respiratory fluoroquinolones, and several studies have reported a potential link between the use of fluoroquinolones and the occurrence of C. difficile infection. After reviewing the available published data on this topic the evidence for a link seems to be suggestive but not conclusive. Infection-control issues seem to play a more important role than fluoroquinolone use. PMID- 19303567 TI - Recurrent Clostridium difficile infection: causality and therapeutic approaches. AB - Recurrent diarrhoea after successful treatment of the primary episode is a common complication of Clostridium difficile infection (CDI) and may be increasing in frequency. Evidence suggests that persistent alterations in the indigenous bowel flora and failure to mount an effective antibody response to C. difficile toxins are the main mechanisms whereby recurrent CDIs occur. Treatment of the first recurrence with the same agent used for the treatment of the primary episode is reasonable, but a different approach is needed for patients with multiple CDI recurrences. Prolonged, repeated courses of metronidazole are discouraged, but a practical first option in these patients is to use tapered-dose oral vancomycin followed by a pulsed-dose regimen. Other potential options include probiotic approaches, restoration of the normal flora, immunological approaches, toxin binding approaches, and serial therapy with vancomycin followed by rifaximin. Randomised studies including a sufficient numbers of patients have not yet been conducted for the treatment of recurrent CDI and are needed to help guide the formulation of effective recommendations. PMID- 19303568 TI - Nosocomial and antibiotic-associated diarrhoea caused by organisms other than Clostridium difficile. AB - Most cases of nosocomial and antibiotic-associated diarrhoea are caused by alteration of the physiological gut microflora. This alteration leads to reduced microbial metabolism of carbohydrates and primary bile acids, resulting in osmotic or secretory forms of diarrhoea. Moreover, facultative enteropathogens may experience a growth advantage due to the antibiotic-induced microflora alteration that, in turn, can harm the gut mucosa by the toxins they produce. Clostridium difficile is the major infectious agent leading to pseudomembranous colitis. However, there is increasing evidence that certain other pathogens such as enterotoxin-producing Clostridium perfringens, Staphylococcus aureus and Klebsiella oxytoca can induce mucosal deterioration and diarrhoea after antibiotic use. But, as with C. difficile, these facultative enteropathogens can also be found in the healthy population. Their contribution to disease is, therefore, controversial and their presence in the stools of antibiotic associated diarrhoea patients is often claimed to be mere colonisation. In this respect, the causal relationship of each suspected pathogen with the development of intestinal disease has to be proved clinically and experimentally. PMID- 19303569 TI - Clostridium difficile is no longer just a nosocomial infection or an infection of adults. AB - Clostridium difficile is the main cause of nosocomial gastrointestinal disorders. Historically, C. difficile has usually affected older patients, hospital inpatients, and long-term care facility residents. Recent reports suggest that the occurrence and severity of C. difficile-associated disease (CDAD) is increasing in populations previously considered to be at low risk of the infection, and increasing numbers of community-acquired cases of CDAD are being reported. Risk factors for CDAD in paediatric patients include disruption of the normal microflora of the gastrointestinal tract (antibiotic-associated and non antibiotic-associated), age, immune status, diet, underlying conditions, concurrent infections, and cancer. CDAD in populations previously thought to be at low risk is an emerging problem. PMID- 19303570 TI - New approach to the management of Clostridium difficile infection: colonisation with non-toxigenic C. difficile during daily ampicillin or ceftriaxone administration. AB - Non-toxigenic strains of Clostridium difficile are highly effective in preventing toxigenic C. difficile infection in hamsters when given following a single dose of an antimicrobial agent. The goal of this study was to determine the ability of non-toxigenic C. difficile to colonise hamsters during administration of an antibiotic to which the organisms are resistant - ceftriaxone - and an antibiotic to which they are susceptible - ampicillin - and to determine if non-toxigenic colonisation is protective against toxigenic strain challenge. Groups of four or five hamsters were administered daily ceftriaxone 60 mg/kg/d intraperitoneally or ampicillin 60 mg/kg/d orally for 5 days. Three non-toxigenic strains of C. difficile, M3, M23, and T7 (MICs 96-128 mg/L) were each given orally at a dose of 1 x 10(6) spores to groups of five animals 3h after the first dose of ceftriaxone. All animals were colonised successfully by day 3 of the study and when challenged with 1 x 10(6) spores of toxigenic strain J9 (MIC >256 mg/L) on day 3 all animals survived, whereas the control animal given ceftriaxone, but not non-toxigenic C. difficile, died within 48h of challenge. When groups of four hamsters were given ampicillin, administration of non-toxigenic strain M3 (MIC 2 mg/L) or toxigenic strain J9 (MIC 0.75 mg/L) at 1 x 10(6) spores did not result in any colonisation or infection of the animals until day 8, 3 days after the last ampicillin dose. A protection study was designed by giving M3 spores to groups of five animals daily for 5 days beginning on day 1, 3, or 5 of ampicillin. Toxigenic challenge was given with J9 spores on day 3 of each M3 regimen. M3 colonised all animals by day 8 and none became infected with J9. Colonisation by non-toxigenic C. difficile is an effective prevention strategy during antibiotic administration of ceftriaxone or ampicillin, but multiple-day administration is required for ampicillin and colonisation does not occur until several days after the drug is discontinued. PMID- 19303571 TI - Alternative strategies for Clostridium difficile infection. AB - Although antibiotics are generally effective in achieving symptomatic recovery from Clostridium difficile infection, the disease frequently relapses, partly because antibiotics not only kill C. difficile, but also disrupt colonisation resistance of the gut microflora. Non-antibiotic strategies for the prevention and treatment of the infection include probiotics, deliberate colonisation by non toxigenic C. difficile strains, toxin-binding agents, active immunisation, passive immunotherapy with intravenous immunoglobulin, monoclonal antibodies or bovine anti-C. difficile whey concentrate, and faecal transplantation. None of these alternative therapies has proven benefit in therapy or prevention, and prospective randomised trials are urgently needed. PMID- 19303572 TI - What is the role of antimicrobial resistance in the new epidemic of Clostridium difficile? AB - The epidemiology of Clostridium difficile infection (CDI) has changed in recent years, with the occurrence of large outbreaks of infection associated with the emergence of 'hypervirulent' strains, particularly PCR ribotype 027. There has been much speculation as to the nature of the factors responsible for driving the new epidemic of CDI, and various hypotheses have been proposed. These include increases in the size of the population at risk of CDI and/or their susceptibility to infection, increased exposure to the organism, and changes in the virulence/ transmissibility of the pathogen. Resistance to a range of antimicrobial agents, including the fluoroquinolones, is a common feature of these newly emerged strains. This article considers the part antimicrobial resistance may play as a driver for the observed changes in the epidemiology of CDI and presents a model that would support such a role. However, the paucity of definitive evidence for a causal role and the complexity of the epidemiology of CDI are acknowledged. It may be more accurate to view antimicrobial resistance as one of a range of factors which interact to alter the parameters of the 'CDI equation', and thereby determine the size of the CDI problem associated with any given emerging subtype of C. difficile. PMID- 19303573 TI - Should angiotensin-converting enzyme inhibitors be used in all patients with coronary artery disease or restricted to those with a history of myocardial infarction or myocardial revascularization? PMID- 19303574 TI - Cardiac rehabilitation provides favourable long-term dietary changes. Why not extend the educational programme? PMID- 19303575 TI - Angiotensin-converting enzyme inhibition with perindopril in patients with prior myocardial infarction and/or revascularization: a subgroup analysis of the EUROPA trial. AB - BACKGROUND: The European trial on Reduction Of cardiac events with Perindopril in patients with stable coronary Artery disease (EUROPA) demonstrated the benefits of perindopril with respect to secondary prevention of cardiovascular risk in patients with stable coronary artery disease. AIMS: To describe the clinical effects of perindopril in a subpopulation of patients from EUROPA with a history of myocardial infarction and/or revascularization. PATIENTS AND METHODS: Of the 12,218 patients in the EUROPA study, 10,962 had a history of myocardial infarction and/or revascularization. In this EUROPA subpopulation, 7910 patients had a history of myocardial infarction and 6709 had a history of revascularization. Patients were randomized to treatment with perindopril 8mg/day or placebo. The primary endpoint was a composite of cardiovascular mortality, myocardial infarction and resuscitated cardiac arrest. RESULTS: After a mean follow-up of 4.2 years, treatment with perindopril 8mg/day was associated with a 22.4% reduction in the primary endpoint compared with placebo (p<0.001) in patients with a history of myocardial infarction. Patients with a history of myocardial revascularization showed a 17.3% reduction in the primary endpoint with perindopril versus placebo (p<0.05). In the combined population of patients with a history of myocardial infarction and/or revascularization, treatment with perindopril produced a 22.4% reduction in the primary endpoint compared with placebo (p<0.001). CONCLUSIONS: This study confirms the benefits of a high dose of angiotensin-converting enzyme inhibitor for the secondary prevention of cardiovascular risk among patients with a history of myocardial infarction and/or revascularization. PMID- 19303576 TI - Sustained positive impact of a coronary rehabilitation programme on adherence to dietary recommendations. AB - BACKGROUND: Nutrition has a major influence after coronary events but long-term adherence to dietary advice is poorly evaluated. AIM: To evaluate if a cardiovascular rehabilitation programme including dietary counselling has a positive impact on adherence to dietary recommendations. METHODS: Two groups of coronary patients were compared in an observational non-randomized study. Group I included 32 patients at the acute phase of a coronary syndrome and group II included 104 patients between six months and three years after completing a cardiovascular rehabilitation programme. The evaluation was performed with (1) a validated 14-item food frequency questionnaire (FFQ), which gives scores for the consumption of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), Omega-6 and Omega-3 polyunsaturated fatty acids (PUFA), fruits and vegetables, and a global cardiovascular protective dietary score; and (2) biological markers. RESULTS: SFA score was higher in group I vs II (7.4+/-2.8 vs 4.4+/-2.1, p<0.001) whereas Omega-3 PUFA (2.2+/-2.0 vs 4.7+/-2.1, p<0.001), fruit and vegetables score (3.3+/-1.4 vs 4.3+/-1.7, p=0.001) and global dietary score (-1.1+/-4.5 to 7.0+/-4.9, p<0.001) were higher in group II. The Omega-6:Omega-3 PUFA ratio was higher in group I (14.2+/-12.7 vs 6.3+/-5.4, p<0.001). Biological markers showed higher plasma contents of Omega-3 PUFA (4.05+/-1.70% vs 2.80+/-1.07%, p<0.001), folate (19.7+/-12.2 nmol/L vs 13.0+/-5.0 nmol/L, p<0.001) and vitamin C (7.60+/ 3.99 mg/L vs 4.18+/-3.46 mg/L, p<0.001), and a higher erythrocyte membrane Omega 3 PUFA content (6.60+/-2.19% vs 5.38+/-2.17%, p=0.016) in group II vs I. CONCLUSION: Using a short FFQ, this study showed sustained improvement in dietary habits in patients with coronary heart disease who receive nutritional education during a cardiovascular rehabilitation programme. PMID- 19303577 TI - Conotruncal defects associated with anomalous pulmonary venous connections. AB - BACKGROUND: Conotruncal defects constitute one of the major categories of congenital heart disease. Our understanding of how these defects develop has been derived from knowledge of the role of neural crest cells in heart development. However, recent studies have revealed a role for the myocardium in the formation of both the arterial and venous poles of the heart. AIM: To identify congenital heart defects that associate anomalies of the arterial and venous poles. METHODS: From a database spanning 27 years, we identified those patients with conotruncal defects associated with an anomalous pulmonary venous connection (APVC; total or partial). Patients with atria isomerism or atrioventricular septal defects were excluded. Patient files were reviewed for clinical presentation, family history, diagnostic and surgical procedures, and outcome. RESULTS: We identified 23 patients with conotruncal defects and APVC. Conotruncal defects were as follows : common arterial trunk, n=7; tetralogy of Fallot, n=5; discordant ventriculoarterial connections, n=4; interrupted aortic arch, n=2; subarterial ventricular septal defect, n=2; double outlet right ventricle, n=2; and right pulmonary artery from ascending aorta, n=1. Nine patients had total APVC and 14 patients had partial APVC. Recurrence of the cardiac defects in siblings was observed in three families. CONCLUSION: Our findings suggest that congenital heart defects that associate anomalies of the arterial and venous poles may have a common embryology, which results from a myocardial defect. PMID- 19303578 TI - Development of catheter-based treatment of patent ductus arteriosus: a medium sized centre experience. AB - BACKGROUND: Despite the availability of effective devices, percutaneous closure of patent ductus arteriosus (PDA) can be challenging in some situations. AIM: To describe our initial experience of percutaneous PDA closure. METHODS: Between 2001 and 2007, 73 consecutive patients aged 3 months to 70 years underwent transcatheter PDA closure. An Amplatzer duct occluder (ADO) was chosen for ducts greater than 2 mm (n=50) and a Detachable coil (DC) for smaller ducts (n=23). RESULTS: The diameter of the ducts ranged from 1 to 7.2 (mean 2.9+/-1.3) mm. The prostheses were implanted successfully in all patients. The complete closure rate reached 98% in the ADO group and 100% in the DC group at 12 months. Four (5.4%) patients showed asymptomatic device protrusion: three patients (5, 6 and 10 kg) into the aortic isthmus and one patient (7 kg) into the pulmonary artery (PA). One patient (7 kg) experienced transient severe bradycardia due to pulmonary air embolism. Another patient (3.3 kg) had a permanent asymptomatic occlusion of the femoral artery. In a third patient (17 kg), the ADO migrated asymptomatically into the descending aorta and was discovered 12 months later. CONCLUSION: Even during the learning curve, percutaneous PDA closure can give excellent results. Strict adherence to protocols and careful follow-up assessments are mandatory. In small infants, the use of the ADO may lead to obstruction in the aorta or PA, or to device migration. Cautious surveillance for untoward events is essential, especially in small infants with large ducts. PMID- 19303579 TI - Risk factors for mortality in patients with mediastinitis after cardiac surgery. AB - BACKGROUND: Patients with mediastinitis after cardiac surgery have higher morbidity and mortality. AIMS: Describe the characteristics of patients with mediastinitis, determine the mortality within one month, and assess the risk factors associated with mortality. METHODS: Retrospective cohort study including all adult patients with mediastinitis during the 2002-2006 period at the Nantes University Hospital. Multivariate analysis by logistic regression and Kaplan Meier curve of survey were done. RESULTS: Nearly 5574 patients were operated during the study period, with a mediastinitis incidence rate of 0.7%, 28 patients (72%) had coronary artery bypass graft. The mortality rate increased from de 12.8% during hospital stay to 20.5% within one year. Only two deaths were associated with mediastinitis. The occurrence of a co-infection was the only independent risk factor associated with mortality (OR 13, P<0.04). The instantaneous risk of death was increased by 7 in patient with co-infection, particularly mechanical ventilator-associated pneumonia (CR 1,97). CONCLUSION: Mortality varied according to the duration of surveillance, and mediastinitis was not the major cause of death. Mechanical ventilator-associated pneumonia after mediastinitis increases the mortality and needs specific prevention. PMID- 19303580 TI - Changes in blood pressure in a large cohort of elderly individuals: Study 3C. AB - OBJECTIVE: Analysis of changes in blood pressure with a two-year interval, and of factors associated with this change, in a large cohort of elderly individuals. METHODS: Follow-up of a cohort of 9294 individuals aged 65 years and over recruited from the general population for Study 3C. Changes in blood pressure are defined as the difference in its averages between the inclusion visit and the follow-up visit at 2 years. The factors associated with changes in systolic blood pressure were identified by univariate and multivariate analyses. RESULTS: Systolic and diastolic blood pressure decreased on average by 7.60 mmHg and 4.45 mmHg respectively in 7659 individuals included in the study between the initial measurement and the follow-up at 2 years. The analyses revealed that the initial high blood pressure level was the main factor for this decrease that would be explained by a phenomenon of regression towards the mean. CONCLUSION: These results confirm the importance of repeating blood pressure measurements during several examinations for a good estimate of individual blood pressure values in this age range. It is also important to consider this phenomenon in studies including specific blood pressure estimates only. PMID- 19303581 TI - Right ventricular apex pacing: is it obsolete? AB - Clinical trials in patients with pacemakers for sinus node dysfunction or atrioventricular block have highlighted the fact that desynchronization of ventricular contraction induced by right ventricular apical pacing is associated with long-term morbidity and mortality. These clinical data confirm pathophysiological results indicating that right ventricular apical pacing causes abnormal ventricular contraction, reduces pump function and leads to myocardial hypertrophy and ultrastructural abnormalities. In this manuscript, we discuss the clinical evidence for the adverse and beneficial effects of various right ventricular pacing sites, left ventricular pacing sites and biventricular pacing. We also propose a decisional algorithm for pacing modalities, based on atrioventricular conduction, left ventricular function and expected lifespan. PMID- 19303582 TI - Intravascular ultrasound-guided percutaneous coronary interventions in contemporary practice. AB - Intravascular ultrasound imaging has been pivotal in the understanding of coronary artery disease and the development of percutaneous coronary intervention. The ability to analyse vessel walls and measure atherosclerotic lesions more accurately has enabled the field of invasive cardiology to overcome the limits of angiography. In fact, intravascular ultrasound measurements correlate with functional measurement of coronary blood flow, as a result interest in their use for the diagnosis of lesion severity in ambiguous lesions and for left main trunk analysis has grown. On the interventional side, intravascular ultrasound is used to determine the major predictors of restenosis and stent thrombosis, which are the main pitfalls of percutaneous coronary intervention. In the bare-metal stent era, intravascular ultrasound-guided percutaneous coronary intervention was associated with a reduction in restenosis rates because it enabled identification and treatment of the risk factors for complications. Although drug-eluting stents have provided a great technological advance in percutaneous coronary intervention, further reducing the rate of in stent restenosis, they have not abolished restenosis completely; intravascular ultrasound has also been used in this setting to identify the mechanisms responsible for drug-eluting stent restenosis. As in the bare-metal stent era, identification of the predictors of restenosis and stent thrombosis and their subsequent treatment may offer the promise of improved outcome in the drug eluting stent era. This review focuses on the potential benefit of intravascular ultrasound-guided percutaneous coronary intervention with regard to restenosis and stent thrombosis in the bare-metal stent and drug-eluting stent eras. PMID- 19303583 TI - Aorto-right ventricular fistula presenting 10 years after aortic surgery as an acute coronary syndrome. PMID- 19303584 TI - Unusual pacemaker location due to pocket infection. PMID- 19303585 TI - Submitral left ventricular aneurysm presenting as a "strange" mitral valve prolapse. PMID- 19303586 TI - Physiological responses to workload change. A test/retest examination. AB - The purpose of this study is to examine the test/retest consistency of physiological responses induced by mental tasks. Fifteen healthy male university students were recruited as participants. They were instructed to perform a 5-min Multi-Attribute Task Battery (MATB) trial three times successively. The task difficulty level of the tracking task of the second trial was set as medium (M). The first one was set as more difficult (H) and the last trial was easiest (L). The difficulty levels of the other two tasks (resource management and system monitoring) of the MATB were identical for all three trials. The participants repeated this procedure on three different days separated by at least a 1-day interval. The order of the tasks was the same for all repeated trials, i.e., H-M L. Tissue blood volume from the tip of the nose using a laser Doppler blood flow meter, skin potential level (SPL), ECG from three leads on the chest, systolic time intervals (pre-ejection period, left ventricular ejection time), and hemodynamic parameters (stroke volume, cardiac output) were recorded during the task trials and before and after 5-min resting periods. The participants reported their subjective workload via NASA-TLX after each task trial. Autonomic nervous system parameters derived from the above-mentioned signals, subjective workload scores, and performance indices of MATB were analyzed, and test/retest reliability was investigated. The results showed that a significant test/retest correlation was obtained for SPL for more participants than in the other parameters, although there were large individual differences. PMID- 19303587 TI - Visual false memories in post-traumatic stress disorder (PTSD). AB - There is an ongoing debate whether or not patients with post-traumatic stress disorder (PTSD) are more prone to produce false memories. The present study investigated this question using a visual variant of the Deese-Roediger-McDermott (DRM) paradigm, additionally addressing underlying mechanisms of false memory production (e.g., depression, dissociation, emotional valence, arousal). The visual paradigm was administered to 48 traumatized individuals with (n=20) and without PTSD (n=28) and 28 non-traumatized controls. Groups did not differ with regard to memory performance and memory confidence. False memories were correlated with depression. We recommend that future studies employ trauma related material to further explore memory aberrations in PTSD. PMID- 19303588 TI - [Mortality risk factors in surgical patients in a tertiary hospital: a study of patient records in the period 2004-2006]. AB - OBJECTIVE: To determine mortality risk factors in surgical patients. MATERIAL AND METHOD: A cross-sectional study was carried out on all surgical patients who died while in hospital, over a period of three years (2004-2006). Pre, intra and postoperative variables were analysed. Comparisons were made between patients operated on as emergencies and elective surgery patients. Multivariate analysis was performed on the pre, intra and postoperative variables, using chi(2) of Pearson correlation with a confidence interval of 95%. RESULTS: Surgery was performed on a total of 38 815 patients, of which 6 326 were emergency procedures and 32 489 as elective. There were 479 deaths registered: 36 occurred in the operating theatre and 443 died after the operation. Arterial hypertension, diabetes mellitus and cancer were significant causes of death. Intraoperative complications were associated with mortality during the surgical procedure. Emergency surgery was an independent risk factor (mortality, 5.5% vs. 0.4% for elective surgery). Sepsis, cardiac and respiratory related deaths were the main risk factors for postoperative death. CONCLUSIONS: Prevention and adequate treatment of perioperative risk factors should significantly reduce morbidity and mortality rates, mainly in those patient operated as emergencies. PMID- 19303589 TI - [Follicular dendritic cell tumour in the anterior mediastinum]. PMID- 19303590 TI - [Anal intraepithelial neoplasia: application of a diagnostic protocol in risk patients using anal cytology]. AB - INTRODUCTION: Anal intraepithelial neoplasia is a precursor condition of squamous anal carcinoma. The groups at risk of this lesion are patients with anogenital condylomata, cervical dysplasia, human immunodeficiency virus infection and, in general, patients with HPV infection. The aim of this study was to analyse the results of a diagnostics protocol of Anal Intraepithelial Neoplasia in high risk population using anal cytology. PATIENTS AND METHOD: The protocol is based on a visit in the outpatient department, clinical interview, physical examination and anal cytology evaluated by Bethesda criteria. The cross-sectional observational study was designed to study the anal smear results and their relationship with risk factors. RESULTS: A total of 64 patients were included from January 2005 to December 2006. In the overall series, 25 patients have been diagnosed with abnormal anal cytology: 9 atypical squamous cells of undetermined significance (ASCUS), 15 low-grade and 1 high-grade squamous intraepithelial lesions. There were no significant associations between abnormal cytology results and the presence of anal condyloma (p = 0.22). Neither were there statistical associations found with high risk-HPV infection (p = 0.84), HIV infection (p = 0.98) or tobacco use (p = 0.14). CONCLUSIONS: Our diagnostic protocol of anal intraepithelial neoplasia revealed 25% of patients with pre-invasive lesions of squamous anal cancer. PMID- 19303591 TI - Twenty four-month-old infants' interpretations of novel verbs and nouns in dynamic scenes. AB - The current experiments address several concerns, both empirical and theoretical in nature, that have surfaced within the verb learning literature. They begin to reconcile what, until now, has been a large and largely unexplained gap between infants' well-documented ability to acquire verbs in the natural course of their lives and their rather surprising failures to do so in many laboratory-based tasks. We presented 24-month-old infants with dynamic scenes (e.g., a man waving a balloon), and asked (a) whether infants could construe these scenes flexibly, noticing the consistent action (e.g., waving) as well as the consistent object (e.g., the balloon) and (b) whether their construals of the scenes were influenced by the grammatical form of a novel word used to describe them (verb or noun). We document that 24-month-olds' representations of novel words are sufficiently precise to permit them to map novel verbs to event categories (e.g., waving events) and novel nouns to object categories (e.g., balloons). We also document the time-course underlying infants' mapping of the novel words. These results beckon us to move beyond asking whether or not infants can represent verb meanings, and to consider instead the conditions that support successful verb learning in infants and young children. PMID- 19303592 TI - The role of the anterior temporal lobes in the comprehension of concrete and abstract words: rTMS evidence. AB - Conceptual knowledge allows us to bring meaning to our world. Studies of semantic dementia (SD) patients and some functional neuroimaging studies indicate that the anterior temporal lobes, bilaterally, are a core neural substrate for the formation of conceptual representations. The majority of SD patients (who have circumscribed atrophy of the anterior temporal lobes) have better comprehension of concrete than abstract words. However, this finding remains controversial, as some individual SD patients have exhibited reverse imageability effects, i.e., relative preservation of abstract knowledge. This would imply that the anterior temporal lobes are particularly crucial for processing sensory aspects of semantic knowledge, which are an important part of concrete but not abstract concepts. To adjudicate on this debate, we used offline, low-frequency, repetitive transcranial magnetic stimulation to disrupt neural processing temporarily in the left or right temporal poles (TPs). We examined this effect using a synonym judgement task, comprising high, medium and low imageability items, which we have previously employed with a case-series of SD patients. The time required to make semantic decisions was slowed considerably, particularly for low imageability items, consistent with the pattern we observed in SD. These results confirm that both TPs make a critical contribution to semantic processing, even for abstract concepts that do not have strong sensory representations. PMID- 19303593 TI - The impact of executive functions on verb production in patients with Parkinson's disease. AB - A growing number of studies suggest that language problems in Parkinson's disease (PD) are a result of executive dysfunction. To test this hypothesis we compared Dutch verb production in sentence context in a group of 28 PD patients with a control group consisting of 28 healthy participants matched for age, gender and education. All subjects were assessed on both verb production in sentence context as well as on cognitive functions relevant for sentence processing. PD patients scored lower than healthy controls on the verb production ability-scale and showed a response pattern in which performance was worse (1) in base than in derived position; (2) in present than in past tense; (3) for intransitive than in transitive verbs. For the PD group the score on the verb production ability-scale correlated significantly with set-switching and working memory. These results provide support for previous research suggesting that executive dysfunctions underlie the performance of the PD patients on verb production. It is furthermore suggested that because of failing automaticity, PD patients rely more on the cortically represented executive functions. Unfortunately, due to the disturbed intimate relation between the basal ganglia and the frontal cortex, these executive functions are also dysfunctional. PMID- 19303594 TI - fMRI studies on drawing revealed two new neural correlates that coincide with the language network. PMID- 19303595 TI - Negative effects of serum p,p'-DDE on sperm parameters and modification by genetic polymorphisms. AB - OBJECTIVE: Effects of ambient exposure to DDT and its metabolites (DDE-DDT) on human sperm parameters and the role of genetic polymorphisms in modifying the association were investigated. METHODS: Demographics, medical history data, blood and semen samples were obtained from the first 336 male partners of couples presenting to 2 infertility clinics. Serum was analyzed for organochlorines (OC) and DNA for polymorphisms in GSTM1, GSTT1, GSTP1 and CYP1A1. Men with each sperm parameter considered low by WHO criteria (concentration <20million/mL, motility <50%, morphology <4%) were compared to men with all normal sperm parameters in logistic regression models, controlling for sum of other OC pesticides. RESULTS: High DDE-DDT level was associated with significantly increased odds for all 3 low sperm parameters. The risk of low motility with high DDE-DDT exposure was increased in men with the GSTT1 null genotype compared to those with GSTT1 intact (odds ratio (OR)=4.19, 95% confidence interval (CI) 1.05-16.78 and OR=3.57, 1.43 8.93, respectively). Risk for low morphology in men with high DDE-DDT and one or both CYP1A1*2A alleles was lower compared to men with the common CYP1A1 alleles (OR=2.18, 0.78-6.07 vs. OR=3.45, 1.32-9.03, respectively). Similar results were obtained for men with low DDE-DDT exposure. Effects of high DDE-DDT on low sperm concentration (OR=2.53, 1.0-6.31) was unaffected by the presence of the polymorphisms. CONCLUSION: High DDE-DDT exposure adversely affected all 3 sperm parameters and its effects were exacerbated by the GSTT1 null polymorphism and by the CYP1A1 common alleles. PMID- 19303597 TI - Experiences of male patients and wife-caregivers in the first year post-discharge following minor stroke: a descriptive qualitative study. AB - BACKGROUND: Most patients with minor stroke are discharged directly home from acute care, under the assumption that little will be required in the way of adaptation and adjustment because informal caregivers will manage the stroke recovery process. We explored male patients with minor stroke and their wife caregivers' perceptions of factors affecting quality of life and caregiver strain encountered during the first year post-discharge. METHODS: Data were obtained from responses to two open-ended questions, part of quality of life and caregiver strain scales administered to participants in a larger descriptive study. Conventional content analysis was used to assess narrative accounts of living with minor stroke provided by 26 male patients and their wife-caregivers over a period of 1-year post-discharge. RESULTS: Two major themes that emerged from these data were 'being vulnerable' and 'realization'. Subthemes that arose within the vulnerability theme included changes to patients' masculine image and wife caregivers' assumption of a hyper-vigilance role. In terms of 'realization' patients and their wife-caregivers shared 'loss' as well as 'changing self and relationships'. Patients in this study focused primarily on their physical recovery and their perceptions of necessary changes. Wife-caregivers were actively involved in managing the day-to-day demands that stroke placed on individual, family and social roles. CONCLUSIONS: We conclude that patients and wife-caregivers expend considerable time and energy reestablishing control of their lives following minor stroke in an attempt to incorporate changes to self and their relationship into the fabric of their lives. PMID- 19303598 TI - WITHDRAWN: Erratum to "Impaired renal vascular endothelial function in vitro in experimental hypercholesterolemia" [ATH 154 (2001) 195-201]. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19303599 TI - Improving the sensitivity of liquid chromatography-tandem mass spectrometry analysis of hexabromocyclododecanes by chlorine adduct generation. AB - It is well documented and experimentally confirmed that hexabromocyclododecanes (HBCDs) tend to associate with several anions forming different adducts that can affect the sensitivity and the accuracy of the determinations. In the present work, two different approaches for HBCD determination have been optimised and characterised based on their repeatability and intermediate precision, linear calibration ranges, sensitivity, limits of detection and quantification and application to commercial food samples. Both methods involve the use of a triple quadrupole mass spectrometer coupled to a liquid chromatograph and the addition of different ammonium salts to the mobile phase, i.e. ammonium chloride or ammonium acetate, in order to encourage (Cl method) or try to inhibit (Ac method), respectively, the formation of the chlorine adducts of the molecular ion. Precision of the two methods investigated was similar and both approaches presented a comparable behaviour for the analysis of food samples. However, the Cl method showed higher sensitivity and the limits of detection (0.23-0.41 pg on column) and quantification (0.77-1.35 pg on column) were up to 14 times lower than those obtained applying the Ac method. All these facts make the Cl method the best choice for the quantification of HBCDs in food samples with low concentration levels. PMID- 19303600 TI - (L)- or (D)-valine tert-butylamide grafted on permethylated beta-cyclodextrin derivatives as new mixed binary chiral selectors: versatile tools for capillary gas chromatographic enantioseparation. AB - This work deals with the synthesis of two mixed binary chiral selectors prepared by grafting (L)- or (D)-valine tert-butylamide on permethylated cyclodextrin macrocycle. The enantioselective properties of the new chiral selectors diluted in OV11 polysiloxane (35% phenyl- and 65% methylsiloxane) were investigated by means of injections of 117 racemic mixtures. The mixed chiral selectors with (L) valine and, to a lesser extent with (D)-valine, were found to have an improved enantioselectivity toward amino acid derivatives by comparison to permethylated cyclodextrin. The enantioseparation capability of these new chiral selectors has proven to be slightly less efficient than Chirasil-L-Val (Alltech) for amino acid derivatives, but it has been extended to include terpenes, lactones, esters, aliphatic compounds and aryl alcohols. PMID- 19303601 TI - Method development for liquid chromatographic/triple quadrupole mass spectrometric analysis of trace level perfluorocarboxylic acids in articles of commerce. AB - An analytical method to identify and quantify trace levels of C5-C12 perfluorocarboxylic acids (PFCAs) in articles of commerce (AOCs) was developed and rigorously validated. Solid samples were extracted in methanol, and liquid samples were diluted with a solvent consisting of 60:40 (v/v) methanol and 2 mM ammonium acetate (NH(4)Ac) aqueous solution. In both cases, the samples were spiked with an isotopically labeled recovery check standard. The samples were concentrated in a nitrogen atmosphere (solid samples only), filtered, and then analyzed by HPLC coupled with a tandem mass spectrometer. Method evaluation included selection of the extraction solvent and the sample preparation solvent used to facilitate sample injection into the analytical system, method comparison for extraction and sample concentration, determination of extraction efficiency, instrument and method detection limits, and determination of potential sample loss during filtration and sample storage. Results of consecutive extractions demonstrated that a single extraction step accounts for 70-100% of the "total" PFCAs in the AOCs with the exception of cookware. The instrument's detection limit was < or = 0.05 ng/mL, and the method detection limit were 1.0-3.9 ng/g for solid AOCs and 1.1-6.8 ng/g for liquid AOCs. The method has been used to determine the PFCA content in a wide range of AOCs containing or treated with fluoropolymers and fluorotelomers. PMID- 19303602 TI - Hydration sequence of swelling clays: evolutions of specific surface area and hydration energy. AB - In order to identify the key steps and the driving force for the hydration process of swelling clays, the water adsorption isotherms and enthalpies were measured on monoionic montmorillonite samples saturated with alkali or calcium ions, and on bi-ionic samples saturated with a sodium-calcium mixture. The specific surface area evolution along the hydration process was determined using a recent interpretation of the experimental adsorption isotherms of swelling solids. Results are interpreted in structural terms. Compared with additional data from sample-controlled thermal analysis (SCTA), the results confirm experimentally that the hydration of Li- and Na-montmorillonite is mainly a cation-controlled process, in contrast with the hydration of Cs samples in which the cation contribution to hydration is negligible, as we have already demonstrated using electrostatic calculations or conductivity measurements. PMID- 19303603 TI - Constant vapor pressure emulsions evaporation: Linalool/water stabilized by Laureth 4. AB - Evaporation paths were calculated in the title system using the relevant part of its published phase diagram and the assumption of equilibrium between vapor and liquids. The vapor pressure of linalool was reduced while the emulsion resided in the two-phase region but remained constant within the three-phase range. The duration of the time for constant vapor pressure depended on the relative humidity (RH) to a surprising degree. PMID- 19303604 TI - Antibodies against oxidized LDL and apolipoprotein E polymorphism in demented patients. AB - In serum of 114 patients with dementia and of 102 controls the level of IG class immunoglobulins directed against oxidized LDL and lipids were determined. In isolated DNA apolipoprotein E gene (APOE) polymorphism was identified. In some individuals very high levels of the antibodies were observed. exceeding the 90 percentile in the investigated group. The prevalence of very high anti-ox LDL antibodies level was significantly more frequent in the carriers of epsilon2 allele and less frequent in the carriers of epsilon4 allele. PMID- 19303605 TI - Differences in retinal nerve fiber layer atrophy between multiple sclerosis subtypes. AB - OBJECTIVES: To determine whether retinal nerve fiber layer (RNFL) atrophy in the afferent visual pathway may complement existing tools used to describe and characterize differences across MS subtypes. METHODS: Optical coherence tomography-measured RNFL values were compared over two years in 35 patients (70 eyes) with optic neuritis (ON) as a clinically isolated syndrome (CIS); 39 patients (78 eyes) with relapsing remitting MS (RRMS); and 7 patients (14 eyes) with secondary progressive MS (SPMS). RESULTS: RNFL comparisons involving eyes without ON yielded greater differences between MS subtypes than ON-affected eyes. Overall RNFL values in non-affected eyes were reduced in SPMS patients (83.4 microm), relative to CIS (101.2 microm) (p=0.0009), and RRMS patients (103.7 microm) (p=0.001); and temporal RNFL atrophy was greater in RRMS (64.4 microm) eyes as compared to CIS eyes (73.2 microm) (p=0.02). In ON-affected eyes, RNFL atrophy was greater in SPMS patients (39.5 microm) than CIS patients (58.1 microm) (p=0.03), and in RRMS patients (48.2 microm) relative to CIS patients (p=0.05). RNFL values did not change significantly for any MS subtype during the two-year duration of the study. INTERPRETATION: RNFL thickness may represent a structural marker, which can help distinguish MS subtypes, because the extent of atrophy is commensurate with disease progression. RNFL comparisons between non affected eyes revealed greater differences between CIS, RRMS, and SPMS patients relative to ON-affected eyes, because the impact of prior ON may supplant the effects of disease subtype. PMID- 19303606 TI - Zoobotryon verticillatum Della Chiaje, 1822 (Bryozoa), a new occurrence in the archipelago of the Azores (North-Eastern Atlantic). AB - The new occurrence of the bryozoan Zoobotryon verticillatum Della Chiaje, 1822 is herein recorded in multiple places throughout Azores archipelago. Where introduced, this species has caused important ecological and economical damage and therefore is regarded as invasive. In the Azores, no detrimental effects have yet been noted. The species is so far restricted to marinas (Horta, Faial Island; Vila Franca do Campo, Sao Miguel Island) and a natural coastal pool located near a secondary harbour (Lajes do Pico, Pico Island). A total of 29 colonies were counted during a specific survey conducted in August 2008 in the marina of Horta. The distribution of the species throughout the eastern and central island groups denotes a wide dispersion area and offers control or eradication measures a low probability of success together with the lack of harbour management practices that could effectively prevent the arrival, settlement and dispersal of non native species transported by human-assisted means. PMID- 19303607 TI - Agricultural lands are hot-spots for annual runoff polluting the southern Great Barrier Reef lagoon. AB - The world's largest coral reef ecosystem, the Great Barrier Reef (GBR), continues to be degraded from land-based pollution. Information about the source of pollutants is critical for catchment management to improve GBR water quality. We report here on an 11-year source to sea study of pollutant delivery in runoff from the Fitzroy River Basin (FRB), the largest GBR catchment. An innovative technique that relates land use to pollutant generation is presented. Study results indicate that maximum pollutant concentrations at basin and sub-catchment scales are closely related to the percentage area of croplands receiving heavy rain. However, grazing lands contribute the majority of the long-term average annual load of most common pollutants. Findings suggest improved land management targets, rather than water quality targets should be implemented to reduce GBR pollution. This study provides a substantial contribution to the knowledge base for the targeted management of pollution 'hot-spots' to improve GBR water quality. PMID- 19303608 TI - Impacts of blending on dilution of negatively buoyant brine discharge in a shallow tidal sea. AB - A fine-resolution three-dimensional hydrodynamic model is applied to study the dilution of desalination brine discharged into a tidal sea. Based on given inflow rate and salinity excess of discharge brine, this study explores variations in mid-field dilutions when other low-salinity wastewater is added to the discharge. Findings reveal that this blending leads to a decrease in dilution in the mixing zone and therefore to higher levels of pollutants in this zone, while, on the other hand, the mixing zone occupies a smaller area. The reason is that the discharge of brine creates a density-driven flow that operates to partially remove effluent from the discharge location. This removal is less efficient for the decrease in density excess of the discharge. Hence, in an ambient sea of moderate mixing, blending can be expected to increase the risk of marine pollution in the mixing zone. PMID- 19303609 TI - Recovery of interior brackish marshes seven years after the chalk point oil spill. AB - Seven years after the April 2000 spill of 140,000 gallons of a mixture of No. 6 and No. 2 fuel oils in the Patuxent River, Maryland, heavily oiled brackish marshes showed continuing effects. Stem density and stem height were significantly lower in oiled versus unoiled sites for Spartina alterniflora but not Spartina cynosuroides habitats. In contrast, belowground biomass was significantly lower in S. cynosuroides habitats but not S. alterniflora habitats. Total PAH concentrations were up to 453 mg/kg in surficial soils (0-10 cm) and 2921 mg/kg with depth (10-20 cm). The oil had lost 22-76% of its initial PAH content after seven years, although the oil in marsh soils has undergone little to no additional weathering since Fall 2000. Based on amphipod acute toxicity tests and sediment quality guidelines, 25% of the soils in the marsh are expected to be toxic (ESB-TU(FCV) values>3.0; P(Max)>0.65). PMID- 19303610 TI - Molecular characterizations of chloramphenicol- and oxytetracycline-resistant bacteria and resistance genes in mariculture waters of China. AB - In order to gain an understanding of the diversity and distribution of antimicrobial-resistant bacteria and their resistance genes in maricultural environments, multidrug-resistant bacteria were screened for the rearing waters from a mariculture farm of China. Both abalone Haliotis discushannai and turbot Scophthalmus maximus rearing waters were populated with abundant chloramphenicol resistant bacteria. These bacteria were also multidrug resistant, with Vibriosplendidus and Vibriotasmaniensis being the most predominant species. The chloramphenicol-resistance gene cat II, cat IV or floR could be detected in most of the multidrug-resistant isolates, and the oxytetracycline-resistance gene tet(B), tet(D), tet(E) or tet(M) could also be detected for most of the isolates. Coexistence of chloramphenicol- and oxytetracycline-resistance genes partially explains the molecular mechanism of multidrug resistance in the studied maricultural environments. Comparative studies with different antimicrobial agents as the starting isolation reagents may help detect a wider diversity of the antimicrobial-resistant bacteria and their resistance genes. PMID- 19303611 TI - Stable isotopes and metal contamination in caged marine mussel Mytilus galloprovincialis. AB - Metal concentrations and isotopic composition were measured in different tissues of the mussel Mytilus galloprovincialis in waters of the Balearic Islands (Western Mediterranean) in order to assess pollution levels. The isotopic composition was correlated with lead, cadmium, selenium and nickel obtained from the digestive gland and foot of the mussels. Significant negative correlations were found between cadmium, selenium and zinc and the mussel foot, mainly for (13)C. Significant correlations were also found between lead and cadmium and the digestive gland. Pearson correlations indicated that the (13)C isotopic signal in foot is a good proxy for the concentration of metals such as lead, cadmium, selenium and zinc. Similarly, (15)N isotopic signatures in the digestive gland reflected the lead and cadmium concentration. PMID- 19303612 TI - [Etiology and clinical management of anemia in heart failure. GESAIC Study]. AB - BACKGROUND AND OBJECTIVE: Studies about anemia in heart failure (HF) tend to link the anemia to a cardio-renal dysfunction, and its syndromic value is seldom evaluated. Our objective was to assess the etiology and clinical management of anemia in HF patients in a hospital setting. PATIENTS AND METHOD: Initial cross sectional analysis of a multi-center and prospective cohort of patients with HF and anemia. Anemia was defined according to the WHO criteria; the Modification of Diet in Renal Disease equation was used to assess glomerular filtration and the etiology of anemia was defined according to common criteria. RESULTS: We evaluated 228 patients, with a median age of 79.1 years and 59.65% women. Iron deficiency anemia was present in 36,8% of patients and anemia of chronic disease in 30.3%. Of note, 12.7% cases did not meet any etiological criteria. The main factor associated with iron deficiency was anti-platelet therapy (OR=1.99; 95% CI, 1.16-1.68) and the main factors associated with anemia of chronic disease were the use of angiotensin converting enzyme inhibitors (ACEI) or angiotensin II receptor antagonists (ARA-II) (OR=3.29; 95% CI, 1.36-7.94). The main factor associated with undefined anemia was initial heart failure (OR=5.41; 95% CI, 1.65 17.65). On the other hand, 8.1% of patients required transfusion, 6% were treated with erythropoietin and 25.3% were treated with iron. Both age (OR=1.04; 95% CI, 1-1.08) and hemoglobin level at admission (OR=1.81; 95% CI, 1.46-2.25) were associated with active treatment for anemia. CONCLUSIONS: A clinical study of anemia in patients with HF can establish an etiological diagnosis in 70% of cases, resulting in a more effective treatment. PMID- 19303613 TI - [Tinnitus treatment: neurosurgical management]. AB - Tinnitus is a very frequent symptom affecting 10% of the general population. It corresponds to the perception of an internal noise that can severely impair the quality of life. Tinnitus management requires a multidisciplinary approach in which neuromodulation and neurosurgery tend to play major roles. Classification of tinnitus separates objective tinnitus (i.e., tinnitus that can be heard or recorded) from the more frequent subjective tinnitus (i.e., tinnitus only perceived by the patient). Objective tinnitus is either pulsatile synchronous with heartbeat or asynchronous. In the former, appropriate radiological testing should search for a vascular abnormality as well as other neurological diseases (intracranial hypertension, Arnold-Chiari malformation, vascular loops, etc.). Asynchronous objective tinnitus generally corresponds to muscular contractions that require specific management. The pathophysiology of subjective tinnitus is more complex, showing strong analogies with postamputation pain syndromes. After peripheral middle ear or inner ear damage, auditory deafferentation could result in hyperactivity and/or functional reorganization within central auditory and nonauditory structures. This could explain the persistence of tinnitus after total hearing amputation (e.g., translabyrinthine approach for vestibular schwannoma) and associated symptoms such as hyperacusis or anxiety and depression. This central model finds strong support in animal experiments and in functional neuroimagery (PET, fMRI, MEG). Since no etiologically based therapies are currently available, severe subjective tinnitus management only targets tinnitus tolerance with sound enrichment or cognitive behavior therapy. However, in the near future better knowledge of tinnitus pathophysiology and innovative therapeutic tools could emerge from neuromodulation techniques such as repeated transcranial magnetic or epidural electric stimulation. PMID- 19303614 TI - Pregnane glycosides from Hoodia gordonii. AB - Hoodia gordonii is a 'weight loss' herb, which has gained popularity in the western countries as an appetite suppressant dietary supplement. Phytochemical study of its aerial parts led to isolation of seven pregnane glycosides (hoodigosides W-Z, hoodistanalosides A-B). Their structures were elucidated by chemical degradation studies and spectroscopic methods, including 1D and 2D NMR and CD spectroscopic methods. PMID- 19303615 TI - A preliminary study on the suitability of Cervidil to induce cervical dilation for artificial insemination in ewes. AB - The aim of this study was to evaluate the efficacy of Cervidil, a prostaglandin E(2) (PgE(2))-releasing vaginal insert used for induction of cervical ripening and labour in women, to enhance the ease of transcervical artificial insemination (TCAI) in anestrous ewes (June). It was hypothesized that the use of Cervidil prior to AI would cause dilation of the cervix, and thus alleviate the difficulty associated with traversing the cervix for semen deposition in sheep. Cervidil was inserted 12h before insemination in six Rideau-Arcott ewes; six ewes served as untreated controls. Semen was deposited into the uterus of all six treated ewes but TCAI was possible only in four of the six control ewes. It can be concluded that Cervidil facilitated transcervical semen deposition in anestrous ewes. The treatment with Cervidil has the promise of a technique to improve transcervical AI and to enable non-invasive embryo transfer procedures in sheep. PMID- 19303616 TI - WITHDRAWN: Ultrasonic characterization of formation damage. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19303617 TI - WITHDRAWN: Cavitation induced Becquerel effect. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19303618 TI - WITHDRAWN: Transient unidirectional acoustic streaming in annular resonators. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19303619 TI - Maternal neutralizing antibodies against a CRF01_AE primary isolate are associated with a low rate of intrapartum HIV-1 transmission. AB - Mother-to-child transmission (MTCT) of HIV-1 provides a model for studying the role of passively acquired antibodies in preventing HIV infection. We determined the titers of neutralizing antibodies (NAbs) against six primary isolates of clades B and CRF01_AE in sera from 45 transmitting and 45 nontransmitting mothers matched for the main independent factors associated with MTCT in Thailand. A lower risk of MTCT, particularly for intrapartum transmission, was associated only with higher NAb titers against the CRF01_AE strain, MBA. The envelope glycoprotein of this strain showed an unusually long V2 domain of 63 amino acids, encoding six potential N-linked glycosylation sites. We provided experimental data indicating that the extended V2 domain contributed to the higher level of resistance to neutralization by mothers' sera in this strain. Taken together the data suggest that some primary isolates with specific properties may be useful indicators for identifying protective antibodies. PMID- 19303620 TI - Inefficient entry of vicriviroc-resistant HIV-1 via the inhibitor-CCR5 complex at low cell surface CCR5 densities. AB - HIV-1 variants resistant to small molecule CCR5 inhibitors such as vicriviroc (VVC) have modified Env complexes that can use both the inhibitor-bound and -free forms of the CCR5 co-receptor to enter target cells. However, entry via the inhibitor-CCR5 complex is inefficient in some, but not all, cell types, particularly cell lines engineered to express CCR5. We investigated the effect of increasing CCR5 expression, and hence the density of the inhibitor-CCR5 complex when a saturating inhibitor (VVC) concentration was present, by using 293 Affinofile cells, in which CCR5 expression is up-regulated by the transcriptional activator, ponasterone. When CCR5 expression was low, the resistant virus entered the target cells to a lesser extent when VVC was present than absent. However, at a higher CCR5 level, there was much less entry inhibition at a constant, saturating VVC concentration. We conclude that the relative decrease in entry of a VVC-resistant virus in some cell types results from its less efficient use of the VVC-CCR5 complex, and that increasing the CCR5 expression level can compensate for this inefficiency. PMID- 19303621 TI - A single bioavailability model can accurately predict Ni toxicity to green microalgae in soft and hard surface waters. AB - The major research questions addressed in this study were (i) whether green microalgae living in soft water (operationally defined water hardness<10mg CaCO(3)/L) are intrinsically more sensitive to Ni than green microalgae living in hard water (operationally defined water hardness >25mg CaCO(3)/L), and (ii) whether a single bioavailability model can be used to predict the effect of water hardness on the toxicity of Ni to green microalgae in both soft and hard water. Algal growth inhibition tests were conducted with clones of 10 different species collected in soft and hard water lakes in Sweden. Soft water algae were tested in a 'soft' and a 'moderately hard' test medium (nominal water hardness=6.25 and 16.3mg CaCO(3)/L, respectively), whereas hard water algae were tested in a 'moderately hard' and a 'hard' test medium (nominal water hardness=16.3 and 43.4 mg CaCO(3)/L, respectively). The results from the growth inhibition tests in the 'moderately hard' test medium revealed no significant sensitivity differences between the soft and the hard water algae used in this study. Increasing water hardness significantly reduced Ni toxicity to both soft and hard water algae. Because it has previously been demonstrated that Ca does not significantly protect the unicellular green alga Pseudokirchneriella subcapitata against Ni toxicity, it was assumed that the protective effect of water hardness can be ascribed to Mg alone. The logK(MgBL) (=5.5) was calculated to be identical for the soft and the hard water algae used in this study. A single bioavailability model can therefore be used to predict Ni toxicity to green microalgae in soft and hard surface waters as a function of water hardness. PMID- 19303622 TI - An experimental study of low concentration sludge settling velocity under turbulent condition. AB - Particle Image Velocimetry (PIV) was used to study the settling of activated sludge flocs under turbulent flow conditions. Experimental results showed that a larger particle diameter led to a higher settling velocity while the higher turbulence intensity led to lower settling velocity. Based on the measurements a mathematical relation has been derived which correlates the settling velocity for individual sludge flocs under turbulent conditions through a modified Vesilind equation. Settling velocity shows a power-type relation to sludge particle diameter and an exponential-type relation with turbulence intensity and sludge concentration. PMID- 19303623 TI - Evaluating the usefulness of dynamic pollutant fate models for implementing the EU Water Framework Directive. AB - The European Water Framework Directive (WFD) aims at achieving a good ecological and chemical status of surface waters in river basins by 2015. The chemical status is considered good if the Environmental Quality Standards (EQSs) are met for all substances listed on the priority list and eight additional specific emerging substances. To check compliance with these standards, the WFD requires the establishment of monitoring programmes. The minimum measuring frequency for priority substances is currently set at once per month. This can result in non representative sampling and increased probability of misinterpretation of the surface water quality status. To assist in the classification of the water body, the combined use of monitoring data and pollutant fate models is recommended. More specifically, dynamic models are suggested, as possible exceedance of the quality standards can be predicted by such models. In the presented work, four realistic scenarios are designed and discussed to illustrate the usefulness of dynamic pollutant fate models for implementing the WFD. They comprise a combination of two priority substances and two rivers, representative for Western Europe. PMID- 19303624 TI - Early Eocene primates from Gujarat, India. AB - The oldest euprimates known from India come from the Early Eocene Cambay Formation at Vastan Mine in Gujarat. An Ypresian (early Cuisian) age of approximately 53Ma (based on foraminifera) indicates that these primates were roughly contemporary with, or perhaps predated, the India-Asia collision. Here we present new euprimate fossils from Vastan Mine, including teeth, jaws, and referred postcrania of the adapoids Marcgodinotius indicus and Asiadapis cambayensis. They are placed in the new subfamily Asiadapinae (family Notharctidae), which is most similar to primitive European Cercamoniinae such as Donrussellia and Protoadapis. Asiadapines were small primates in the size range of extant smaller bushbabies. Despite their generally very plesiomorphic morphology, asiadapines also share a few derived dental traits with sivaladapids, suggesting a possible relationship to these endemic Asian adapoids. In addition to the adapoids, a new species of the omomyid Vastanomys is described. Euprimate postcrania described include humeri, radii, femora, calcanei, and tali, most of which show typical notharctid features and are probably attributable to asiadapines. Anatomical features of the limb elements indicate that they represent active arboreal quadrupedal primates. At least one calcaneus is proximally shorter and distally longer than the others, resembling eosimiids in this regard, a relationship that, if confirmed, would also suggest an Asian or southeast Asian faunal connection. Isolated teeth from Vastan Mine recently attributed to a new eosimiid, Anthrasimias gujaratensis, appear to provide that confirmation. However, their attribution to Eosimiidae is equivocal. They are similar to teeth here tentatively referred to Marcgodinotius, hence A. gujaratensis may be a junior synonym of M. indicus. Corroboration of eosimiids at Vastan requires more compelling evidence. Although definitive conclusions are premature, available evidence suggests that the Vastan adapoids, at least, were derived from western European stock that reached India near the Paleocene-Eocene boundary. PMID- 19303625 TI - Bioactivity of POPs and their effects in mosquitofish in Sydney Olympic Park, Australia. AB - The site of the 2000 Olympic Games (Sydney Olympic Park (SOP), Sydney, Australia) was contaminated by persistent organic pollutants (POPs) prior to remediation in the 1990s. This study investigates the bioactivity of POPs in the sediment and water of wetlands across SOP by in vitro 2,3,7,8-TCDD equivalence (TCDDeq) measurement (H4IIE cell line bioassay). Further, it examines whether disturbance of these sediments is likely to mobilise ligands for this receptor into the water column. Exposure to aryl hydrocarbon receptor (AhR) ligands was measured in vivo using hepatic cytochrome P4501A (CYP1A) induction (EROD) in the mosquitofish (Gambusia holbrooki). Aqueous TCDDeq ranged from 0.013 to 0.057 pM in SOP wetlands which was significantly (p<0.05) less that in urban reference sites. These concentrations were not correlated to physical or chemical characteristics of the wetlands. In the sediments, TCDDeq ranged from 0.0016 to 7.06 microg/kg and these were not significantly (p>or=0.05) different to that measured in urban reference sites. Simulated disturbance of small quantities of sediment in water samples significantly (p<0.05) increased the levels of TCDDeq measured in the water. Sediment TCDDeq was correlated to sediment SigmaPAH concentration in 2006 and sediment SigmaPCB, SigmaDDT concentrations and fine sediment grain size in 2005. While fish at one SOP wetland had hepatic EROD activity elevated above the estimated basal level for this species, these were at the lower end of the range measured in urban impacted, non-remediated wetlands. EROD activity was positively correlated with both the sediment SigmaPCB load and aqueous TCDDeq. Increased catchment size was correlated with increased EROD activity suggesting an even spread of POPs throughout the residential areas of the Sydney metropolitan area. The concentration of bioactive POPs in the wetlands of SOP is therefore low relative to urban reference sites demonstrating the ongoing success of the remediation program. PMID- 19303626 TI - Health effects of exposure to carbon nanofibers: systematic review, critical appraisal, meta analysis and research to practice perspectives. AB - BACKGROUND: Literature reviews examining the relationship between exposure to carbon nanofibers (CNFs) and health consequences are qualitative in nature and do not employ an evidence-based assessment. OBJECTIVE: This research deals with a systematic review, critical appraisal, and meta-analysis designed to examine the potential health effects associated with exposure to CNFs. The utilization of research findings into practice is also explored. METHODS: Published articles were obtained from a search of electronic databases and bibliographies of identified articles. A critical appraisal was conducted using an 'Experimental Appraisal Instrument' developed in this study. The meta-analysis was established using statistical techniques with/without the incorporation of overall study quality. The likelihood of utilizing research findings into practice (i.e., from research to practice) was computed using a four-step algorithm based on the criteria of: strength of association, consistency among studies, temporality, biological gradient, type of experimental unit, type of CNF (single- and multi wall nanotubes), CNF grade (commercial or altered), exposure dose, exposure duration, and support by analogy from the published literature. RESULTS: Twenty one experimental studies satisfied the inclusion criteria and were performed on human cells, experimental animal models and animal cells as experimental units. The methodological qualities of published studies ranged from 'very poor' to 'excellent', with 'overall study description' scoring 'good' and 'study execution' equal to 'moderate'. The random-effects model was applied in the meta analysis calculations as heterogeneity was significant at the 10% for all outcomes reported. The mean standardized meta-estimates for the experimental groups were significantly lower than those for the control groups for cell viability and cell death, respectively. Incorporating the effect of overall study quality score widened the gap between the experimental and control groups. Assessment of research findings on the basis of the four-step algorithm revealed that the likelihood of the results to occur in practice is 'somewhat possible' at this time. That is, if exposure conditions to CNF in the reported studies are similar to those in nano-manufacturing plants, it is somewhat possible that CNFs alter the function of human cells resulting in loss of cell viability and cell death. CONCLUSIONS: Our findings suggest that it is 'somewhat possible' for the CNF to penetrate the human cells in the targeted organs and to cause cellular damage. Although the weight of evidence is not sufficient, it is advisable that actions be taken to ensure the protection of workers exposed to CNFs, that is, (a) engineering controls should be established to contain exposure to CNF, and (b) simultaneously rigorous personnel protective equipment should be planned to further minimize the risk of CNF exposure. PMID- 19303627 TI - Anti-prothrombin antibodies are associated with thrombosis in children. AB - INTRODUCTION: This investigation aimed to evaluate thrombotic risk factors in children, with special reference to autoantibodies against prothrombin and protein S. MATERIALS AND METHODS: We studied 57 consecutive Swedish children and adolescents referred with a radiologically confirmed acute thrombotic event. Clinical data were collected and a thrombophilia investigation was performed, including analysis of autoantibodies against protein S (anti-PS) and prothrombin (anti-PT). The anti-PS and anti-PT autoantibodies were also investigated in sera from 47 healthy controls. Detection of autoantibodies was performed by quantitative enzyme-linked immunosorbent assays. RESULTS: Results for anti-PT antibodies were positive in 21% (12/57) of the patients and 2.1% (1/47) of the controls (OR 12.0, 95% CI 1.7-534; p=0.005). Seven percent (4/57) of the patients and 2.1% (1/47) of the controls were positive for anti-PS antibodies (OR 3.4, 95% CI 0.3-174; p>0.30). The FV G1691A mutation was found in 25% (14/57), and 44% (25/57) had 2 or more prothrombotic risk factors. Sixty percent (34/57) of the thrombosis patients were female. Peaks in frequency of thromboembolic events were found in the neonatal and the adolescent periods. Fifty-three percent (30/57) had thrombosis in the lower venous system. Associated clinical conditions occurred in 91% (52/57): systemic illness in 31% (18/57), infections in 26% (15/57), and oral contraceptive use in 25% (14/57). Four percent (2/57) had no apparent clinical or prothrombotic risk factors. CONCLUSIONS: This study suggests that anti-PT autoantibodies may be common risk factors for thrombosis in children, and it confirms the multifactorial nature of pediatric thrombosis. PMID- 19303628 TI - An immortalized goat mammary epithelial cell line induced with human telomerase reverse transcriptase (hTERT) gene transfer. AB - Although mammary epithelial cell lines can provide a rapid and reliable indicator of gene expression efficiency of transgenic animals, their short lifespan greatly limits this application. To provide stable and long lifespan cells, goat mammary epithelial cells (GMECs) were transduced with pLNCX2-hTERT by retrovirus-mediated gene transfer. Transduced GMECs were evaluated by reverse transcriptase polymerase chain reaction (RT-PCR), proliferation assays, karyotype analysis, telomerase activity assay, western blotting, soft agar assay, and injection into nude mice. Non-transduced GMECs were used as a control. The hTERT-GMECs had higher telomerase activity and extended proliferative lifespan compared to non transfected GMECs; even after Passage 50, hTERT-GMECs had a near diploid complement of chromosomes. Furthermore, they did not gain the anchorage independent growth property and were not associated with a malignant phenotype in vitro or in vivo. PMID- 19303629 TI - Elemental ion release from four different fixed prosthodontic materials. AB - OBJECTIVE: Elemental release is important because it plays a significant role in material biocompatibility. The aim of this study was to investigate the release of metal ions from four fixed prosthodontic materials. METHODS: Specimens were prepared using the conventional lost wax technique for gold and nickel-chromium (Ni-Cr) alloys, and by cutting blocks and bar for CAD-CAM ceramic and stainless steel (St-St) alloy, respectively. All specimens were polished (600grit SiC paper), and ultrasonically cleaned with ethanol for 5min. After they were immersed in 0.9% sodium chloride (NaCl) and 1% lactic acid, and were kept at 37 degrees C for 7 days, the elemental release (mug/cm(2)) from each material was analyzed by using inductively coupled plasma mass spectroscopy. The rate (n=5) was statistically analyzed by ANOVA/Tukey test (p<0.05). Two immersion blank solutions were used as a negative control group. RESULTS: Higher elemental release (mean+/-S.D.) of all elements from all materials was evident into the lactic acid solution except for Ag. In the gold alloy, there was significant difference (p<0.05) between Zn and other released elements in the NaCl solution, and it also revealed significant difference between Pd or Ag and Cu which detection value was more than Zn (but no statistical difference) into lactic acid solution. The Ni was significantly more released from Ni-Cr alloy than the other elements into both NaCl and lactic acid solutions. The same was observed for Fe released from St-St alloy. There was more significant release of K than Al from CAD-CAM ceramic in only NaCl solution. SIGNIFICANCE: Transient exposure of tested materials to an acidic environment is likely to significantly increase the elemental release from them. The significant higher release of Ni from Ni-Cr alloy, and Zn, Cu from gold alloy was evident. PMID- 19303630 TI - AIDS: lessons learnt and myths dispelled. PMID- 19303631 TI - PCI or CABG in coronary artery disease? PMID- 19303632 TI - Immunisation timing: the protective layer in vaccine coverage. PMID- 19303633 TI - Timing of children's vaccinations in 45 low-income and middle-income countries: an analysis of survey data. AB - BACKGROUND: Vaccinations are often delayed until well after the recommended ages, leaving many children exposed for longer than they should be. We estimated vaccination coverage at different ages, and delays in administration, in 45 low income and middle-income countries. METHODS: We used data for 217 706 children from Demographic and Health Surveys between 1996 and 2005 (median 2002), which provided data for vaccination of children on the basis of events recorded on vaccination cards and interviews with mothers, with imputation of missing values and survival analysis. We devised an index combining coverage and delay. FINDINGS: For vaccinated children, the median of the median delays in the 45 countries was 2.3 weeks (IQR 1.4-4.6) for bacille Calmette-Guerin (BCG); 2.4 weeks (1.2-3.3) for diphtheria, tetanus, and pertussis (DTP1); 2.7 weeks (1.7 3.1) for measles-containing vaccine (MCV1); and 6.2 weeks (3.5-8.5) for DTP3. However, in the 12 countries with the longest delays for each vaccination, at least 25% of the children vaccinated were more than 10 weeks late for BCG, 8 weeks for DTP1, 11 weeks for MCV1, and 19 weeks for DTP3. Variation within countries was substantial: the median of the IQRs in the 45 countries for delay in DTP3 was 10.9 weeks, 7.9 weeks for MCV1, 5.4 weeks for BCG, and 5.3 weeks for DTP1. The median of the national coverage rates for DTP1 increased from 57% in children aged 12 weeks to 88% at 12 months, and for DTP3 from 65% at 12 months to 76% at 3 years. INTERPRETATION: The timeliness of children's vaccination varies widely between and particularly within countries, and published yearly estimates of national coverage do not capture these variations. Delayed vaccination could have important implications for the effect of new and established vaccines on the burden of disease. FUNDING: WHO's Initiative for Vaccine Research. PMID- 19303634 TI - Coronary artery bypass surgery compared with percutaneous coronary interventions for multivessel disease: a collaborative analysis of individual patient data from ten randomised trials. AB - BACKGROUND: Coronary artery bypass graft (CABG) and percutaneous coronary intervention (PCI) are alternative treatments for multivessel coronary disease. Although the procedures have been compared in several randomised trials, their long-term effects on mortality in key clinical subgroups are uncertain. We undertook a collaborative analysis of data from randomised trials to assess whether the effects of the procedures on mortality are modified by patient characteristics. METHODS: We pooled individual patient data from ten randomised trials to compare the effectiveness of CABG with PCI according to patients' baseline clinical characteristics. We used stratified, random effects Cox proportional hazards models to test the effect on all-cause mortality of randomised treatment assignment and its interaction with clinical characteristics. All analyses were by intention to treat. FINDINGS: Ten participating trials provided data on 7812 patients. PCI was done with balloon angioplasty in six trials and with bare-metal stents in four trials. Over a median follow-up of 5.9 years (IQR 5.0-10.0), 575 (15%) of 3889 patients assigned to CABG died compared with 628 (16%) of 3923 patients assigned to PCI (hazard ratio [HR] 0.91, 95% CI 0.82-1.02; p=0.12). In patients with diabetes (CABG, n=615; PCI, n=618), mortality was substantially lower in the CABG group than in the PCI group (HR 0.70, 0.56-0.87); however, mortality was similar between groups in patients without diabetes (HR 0.98, 0.86-1.12; p=0.014 for interaction). Patient age modified the effect of treatment on mortality, with hazard ratios of 1.25 (0.94-1.66) in patients younger than 55 years, 0.90 (0.75-1.09) in patients aged 55-64 years, and 0.82 (0.70-0.97) in patients 65 years and older (p=0.002 for interaction). Treatment effect was not modified by the number of diseased vessels or other baseline characteristics. INTERPRETATION: Long-term mortality is similar after CABG and PCI in most patient subgroups with multivessel coronary artery disease, so choice of treatment should depend on patient preferences for other outcomes. CABG might be a better option for patients with diabetes and patients aged 65 years or older because we found mortality to be lower in these subgroups. PMID- 19303635 TI - Myelodysplasia or myelodysplastic syndrome? PMID- 19303636 TI - Profiles of school adaptation: social, behavioral and academic functioning in sexually abused girls. AB - OBJECTIVES: The short-term outcomes of child sexual abuse (CSA) on academic, behavioral and social adaptation at school were examined in order to: (1) document the proportion of sexually abused (SA) girls struggling in school and define the nature of their difficulties, (2) explore whether different profiles of school adaptation could be identified, and (3) examine the different constellations of contextual, personal, and familial correlates of abuse for each profile. METHOD: The sample consisted of 100 French-speaking mother-daughter dyads. Girls (aged 7-12, M= 8.64) were administered cognitive functioning tests and completed self-report measures to assess symptomatology, perception of abuse, and support provided. Mothers and teachers completed questionnaires to assess child's emotional, social, behavioral, and academic functioning in school. RESULTS: Descriptive analyses showed that a substantial number of girls (54%) presented clinical school adaptation difficulties in at least one of the three domains evaluated. Cluster analysis revealed four distinctive school functioning profiles: (1) academic-specific, (2) acting out/withdrawn, (3) polyclinical, and (4) resilient. These were distinguished by degree of school functioning difficulties and number of domains affected. The profiles were further discriminated by elements of the CSA experience and contextual, familial, and personal factors. The number of spheres of functioning affected was proportional to the number of risk factors to which girls were exposed. DISCUSSION: Results suggest the need for a systematic and thorough assessment of SA child functioning at school and may guide mental health professionals in providing differential treatment on the basis of the emerged typology. PRACTICE IMPLICATIONS: Sexually abused children demonstrate a wide variability of responses, rendering the elaboration of a "one size fits all" treatment rather difficult. The emerged typology underscores the complex heterogeneity of school adjustment profiles and suggest that secondary effects of abuse on child's level of school functioning require specific assessment and close monitoring. This study raises practical questions as to whether such secondary effects could best be addressed as a primary treatment objective, or in addition to a trauma-focused approach, or as the sole treatment objective. PMID- 19303637 TI - When is a capitally charged defendant incompetent to stand trial? AB - Competence to stand trial is a functional test rather than a bright line test, which therefore requires a case and fact specific assessment of a client's abilities in context. This article discusses competence in the context of capital trial cases. There are serious potential pitfalls for the client when raising incompetence and the decision to do so must be based on the specific ways in which the client's mental illness interferes with specific abilities to communicate with counsel and understand the proceedings. This article addresses counsel's duties in the context of assessing competence, but focuses on the little addressed issue of what abilities a client must have and what tasks a client must participate in so as to be engaged in a competent manner. It also discusses the types of conditions which may interfere with competence to stand trial. PMID- 19303638 TI - Prevalence and demographic correlates of intimate partner violence in Asian Americans. AB - This study provides the first national estimates of the prevalence and correlates of intimate partner violence (IPV) among Asian Americans. Population estimates are based on data from 1470 Asian Americans interviewed for the National Latino and Asian American Study. Interviews were conducted in English, Chinese, Tagalog, or Vietnamese. Results suggest that rates of IPV among Asian Americans are low compared to the general U.S. population. Minor violence victimization by a current intimate partner was reported by 10.2% of women and 12.0% of Asian American men. Notably, a greater proportion of participants admitted having perpetrated IPV than having been a victim. Predictors of IPV included younger age, higher SES, alcohol- and substance-use disorders, depression, ethnicity, and being U.S.-born. Results suggest the need for additional research to examine the interactions between gender, ethnicity, and acculturation to develop group specific models of IPV risk and resilience within diverse Asian American groups. PMID- 19303639 TI - Mental disorder as the cause of a crime. AB - An offender's punishment can be reduced when a court decides that his mental disorder reduces his responsibility for what he did. Courts have sought to establish whether a mentally disordered offender's responsibility is reduced by asking whether his disorder caused the crime. Acceptance of this "causation by mental disorder" criterion has fluctuated, however. This may be because causal explanations are not the types of explanations we are accustomed to offering for the kinds of acts that bring defendants, and psychiatric witnesses, to court. More often, we offer what philosophers have called "possibility" explanations for these acts. The application of psychiatry to possibility explanations has not been widely explored. It offers the potential for the improved use of psychiatric evidence in criminal proceedings. PMID- 19303640 TI - Patients' rights to complain in Finnish psychiatric care: an overview. AB - Cuts in resources for Finnish psychiatric care may jeopardize the realization of patients' rights in mental health settings. The right to complain is a basic right of all patients in Finland, and is especially important to patients treated involuntarily and also to those who have experienced coercive treatment methods during their hospitalizations. In Finland, a patient's right to complain is guaranteed by law, both in legislation and in national quality recommendations. The complaint process in Finland is very complex, and there are several ways to make a complaint that are not always familiar to patients with severe illnesses. Psychiatric patients may have cognitive impairments that make the formulation of a complaint difficult. Despite help from the patient ombudsman, unbalanced power structures in psychiatric hospitals, insufficient information and long evaluation of appeals makes the complaint process very demanding for psychiatric patients. PMID- 19303641 TI - Concentrations of chlorinated and brominated contaminants and their metabolites in serum of harbour seals and harbour porpoises. AB - Harbour seals (Phoca vitulina) and harbour porpoises (Phocoena phocoena) are top predators in the North Sea and consequently accumulate a variety of pollutants in their tissues. Concentrations of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and their hydroxylated metabolites (HO-PCBs and HO-PBDEs) were measured in serum of wild harbour seals (n=47) and captive harbour porpoises (n=21). Both species exhibit long life spans and do not have extreme situations, such as complete fasting during periods of lactation, in their annual cycles. For PCBs, concentrations in adult males were slightly higher than in juveniles and lowest in juvenile females. For PBDEs, juveniles have higher levels than adult males and females, probably as a consequence of lactational transfer. However, differences between these age-gender groups were not statistical significant, indicating that individual variation was limited within each species, even without knowing the feeding status of the animals. Body condition, particularly emaciation, has a major influence on the levels of chlorinated and brominated contaminants in serum. Profiles of PCBs were CB 153>CB 138>CB 187>CB 180 and CB 153>CB 138>CB 149>CB 187>CB 180 for harbour seals and porpoises respectively. For PBDEs, BDE 47 was the predominant congener followed by BDE 100 and 99 in both species. In harbour seals, concentrations of sum PCBs (median: 39,200 pg/ml) were more than 200 times higher than levels of sum PBDEs (median: 130 pg/ml) and almost 10 times higher than concentrations of sum HO-PCBs (4350 pg/ml). In harbour porpoises, concentrations of sum PCBs (median: 24,300 pg/ml) were about 20 times higher than concentrations of PBDEs (median: 1300 pg/ml). HO-PCBs were detected in only 4 harbour porpoises and this at very low concentrations. Naturally-produced MeO-PBDEs were only found in harbour porpoises at concentrations ranging from 120 to 810 pg/ml. HO-PBDEs were not found in any species. In general, harbour seals accumulate less compounds and have mostly lower concentrations than harbour porpoises possibly as a result of a better developed metabolism. PMID- 19303642 TI - The immunoglobulin-binding Eib proteins from Escherichia coli are receptors for IgG Fc. AB - The immunoglobulin-binding proteins from Escherichia coli (Eibs) comprise a family of six proteins homologous to the Yersinia adhesin YadA. These proteins are postulated to bind to the Fc portion of immunoglobulin G (IgG) in a non immune manner. However, a recent study [Ghumra, A., Pleass, R.J., 2007. Escherichia coli do not express Fc-receptors for human immunoglobulin G (IgG). Mol. Immunol. 44, 2144-2146] appeared to show that these proteins do not bind Fc and suggested that the binding seen in earlier studies is due to the polyclonal preparations used in the assays containing antibodies specific to epitopes in the Eib proteins. To resolve this matter, we produced purified, recombinant Eibs for the first time and investigated their binding to intact antibodies and Fc fragments by immunoblot and ELISA techniques. We were able to purify four members of the family, EibA, -C, -D and -F, and show conclusively that these bind IgG Fc. We were also able to block the binding of full-length antibody with IgG Fc, but not with IgG Fab. Binding to IgG Fab was not detectable by surface plasmon resonance, whereas the affinities of Eibs to IgG and IgG Fc were in the range of 50-200 nM. We further demonstrate that deglycosylating IgG Fc does not affect Eib binding. Our results show that the Eib proteins do indeed bind human IgG Fc and that IgG Fc receptors are present in E. coli. PMID- 19303643 TI - WITHDRAWN: Publisher's note. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19303645 TI - Effects of in vitro lactoferricin and lactoferrin on the head kidney cells of European sea bass (Dicentrarchus labrax, L.). AB - Antimicrobial, anti-inflammatory and immunomodulating properties of lactoferrin have been demonstrated in mammals and in fish. However, in vivo, lactoferrin is digested by gastric pepsin treatment into the N-terminal derived peptide named lactoferricin. This has been so far overlooked in fish in vitro studies. The aim of the present study was to assess in vitro the effects of both lactoferricin and lactoferrin on the head kidney cells of European sea bass (Dicentrarchus labrax, L.) in order to determine their potential as dietary additives and to get some insight into their mode of action. In vitro lactoferricin decreased significantly the chemiluminescent response of head kidney cells but did not affect the zymosan triggered chemiluminescence activity. On the other hand, a high concentration of lactoferrin directly stimulated chemiluminescence but reduced the zymosan triggered chemiluminescence. The bactericidal activity of head kidney cells was also significantly diminished by pre-incubation with lactoferrin in a dose dependent manner. Although no significant effect of lactoferricin or lactoferrin was evidenced on head kidney cellular viability, absent or negative effect on the priming of respiratory burst activity suggested that care should be taken when using lactoferrin in the diet of sea bass and high doses should be avoided. Hypotheses about the mechanisms of action of lactoferricin and lactoferrin are presented. PMID- 19303646 TI - Posterior ankyloglossia: a case report. AB - Ankyloglossia, or tongue-tie, refers to an abnormally short lingual frenulum. Ankyloglossia is a recognized but poorly defined condition and has been reported to cause feeding difficulties, dysarthria, dyspnea, and social or mechanical problems. In infants, the most concerning symptoms are feeding difficulties and inability to breastfeed. While a recent trend toward breastfeeding has brought frenulectomy back into favor, the literature regarding treatment remains inconclusive. We report a case of posterior ankyloglossia with anterior mucosal hooding and a simple, safe, and effective way to treat it to improve breastfeeding. PMID- 19303647 TI - Measuring bone speed of sound in patients with hip fractures: a comparison with pathological examination. AB - The aim of this multicenter prospective study is to compare and find a relation between bone strength evaluated by an ultrasound examination (bone speed of sound) and pathological quantitative bone mass measurements, in patients requiring hip replacement due to femoral neck fractures. The study included 93 patients who underwent a bone ultrasound examination to assess bone strength, as well as a pathological quantitative bone mass evaluation of the excised femoral head. Statistically significant correlations were found between the ultrasound results performed at three different sites, between the patient's age and the ultrasound examination results, and between the patient's age and the pathological examination results of the bone strength. We found no statistically significant correlation between the ultrasound examination results in any of the three locations and the pathological results. Therefore, conclusions cannot be made with respect to the efficacy and reliability of the ultrasound examination in predicting bone strength. However, the study can be used as a learning tool for future studies, by trying to overcome the obstacles found in the present study that have most likely lead to the negative results. PMID- 19303648 TI - Gender differences influence the outcome of geriatric rehabilitation following hip fracture. AB - Hip fracture represents the most dramatic expression of the disease, in terms of morbidity, medical cost and mortality. The incidence of hip fracture increases substantially with age. The purpose of this study was to evaluate the association between gender and geriatric rehabilitation outcome after traumatic hip fracture. Ninety-nine community-dwelling elderly patients (older than 65 years), 64 women and 35 men, who were admitted to geriatric rehabilitation after operated hip fracture were studied. We assessed the patients' clinical and demographic data, preoperative risk (ASA), type of fracture and orthopedic repair, pain intensity (VAS), cognitive (MMSE), mood (Zung IDS), and functional status (FIM) on admission and at the end of geriatric rehabilitation. Men had higher mean number of comorbid conditions at the time of the fracture. Men recovered more from depressed mood in comparison with women during the rehabilitation. Significant improvement in FIM motor subscore on discharge was found in both groups. The FIM motor subscore gain was higher in men (24.47) in comparison with women (19.22, p=0.036). Those differences were demonstrated in mean subscores of transfers (p=0.004), and locomotion (p=0.019). Women were more functionally dependent in locomotion, transfers and sphincter control. There were no differences between the groups by duration on rehabilitation stay. Recovery after hip fracture depends in large part on the pre-fracture health and functional ability of the patient. Gender differences in functional recovery may affect therapeutic and rehabilitative decision making. Functional recovery after traumatic hip fracture was better in men in comparison with women. PMID- 19303649 TI - The effects of G-CSF-induced mobilization of progenitor cells are limited by ADMA. AB - OBJECTIVE: Progenitor cells (PC) are thought to induce angiogenesis, and thereby, PC may help to improve ventricular performance in patients with ischemic heart disease (IHD). However, mobilization of progenitor cells by application of G-CSF gives inconsistent clinical effects. The aim of the present study was to assess pathophysiologic effects of progenitor cell mobilization. METHODS AND RESULTS: PC levels (CD34+/CD117+) were counted in 8 patients with severe coronary heart disease and angina pectoris symptoms refractory to conventional therapy during G CSF treatment (5 MUg/kg/d) on days 2, 5, 8, at the end of hospitalization (day 10 12) and after 142+/-33 days of follow-up. Levels of asymmetric dimethylarginine (ADMA; inhibitor of eNOS) and symmetric dimethylarginine (SDMA) were determined at each occasion and correlated with leukocyte count, systemic nitrite levels, myeloperoxidase (MPO) expression in leukocytes, and urine levels of 8-iso prostaglandin F2alpha. Isolated CD34+ cells and endothelial cell cultures were used for functional experiments. G-CSF therapy induced leukocytosis and a rise in CD34+ cell levels. Amounts of MPO positive leukocytes and ADMA levels increased significantly during the treatment phase. ADMA levels correlated to MPO activity (r=0.78; p=0.001) and were inversely related to nitrate levels. In contrast, 8 iso-prostaglandin F2alpha and amounts of SDMA did not change. Culturing endothelial cells in the presence of myeloperoxidase caused an increase in endothelial ADMA synthesis, which was prevented by application of the antioxidant trolox. CONCLUSIONS: Leukocytosis associated with increased MPO activity during G CSF therapy appears to be responsible for the systemic release of ADMA, which impairs eNOS activity. Thus, increased MPO and ADMA levels seem to counteract the potential beneficial effects of PC mobilization. PMID- 19303650 TI - Peripartum cardiomyopathy in type II autoimmune polyendocrine syndrome. AB - Autoimmune polyglandular syndromes (APS) consist of the failure of two or more endocrinal glands together requiring hormone replacement. Type II APS, first described by Schmidt in 1926, is the commonest type of APS. In majority of patients, type II APS consists of Addison's disease in combination with autoimmune thyroid disease and type 1 diabetes. We present a case of APS II in a young lady who developed peripartum cardiomyopathy after an uneventful pregnancy. PMID- 19303651 TI - Let's think in alkaline phosphatase at heart function. AB - In their recent paper, Cheung et al [B.M. Cheung, K.L. Ong, L.Y. Wong, Elevated serum alkaline phosphatase and peripheral arterial disease in the United States National Health and Nutrition Examination Survey 1999-2004. Int J Cardiol 2008 (Electronic publication ahead of print)] described a significant association between serum alkaline phosphatase levels and low ankle-brachial blood pressure index, a risk factor for cardiovascular pathology. We had verified that alkaline phosphatase is present at the rat heart, showing a distribution compatible with cardiomyocyte sarcoplasmic reticulum. Moreover, several drugs with cardiac effect were shown to interfere with heart alkaline phosphatase activity. We therefore propose that alkaline phosphatase may be a local regulator at heart function and a putative target for therapeutic interventions. PMID- 19303653 TI - Bacterial loads and microbial composition in high pressure treated oysters during storage. AB - Analysis of bacterial communities present in high-pressure (HP)-treated, quick frozen (QF), and raw oysters was carried out during three different seasons. Bacterial numbers and species diversity in each sample were determined at 0, 7, 14, and 21 days of storage. Results showed that numbers of total aerobic bacterial counts (TABC) in treated oysters were significantly lower than in untreated oysters at day 0 by 10 to 10(5) colony forming units per gram of oyster meat (CFU/g) in all samplings. However, an increase in TABC in HP-treated oysters was observed at days 7, 14, and 21 indicating that some bacteria survived the treatment and were able to proliferate during refrigeration conditions. Surprisingly, TABC in HP-treated oysters reached 10(8) CFU/g at 14 days of storage in all samplings (higher than TABC from raw oysters in two of the samplings performed). Analysis of the bacterial flora by 16S rDNA sequencing, revealed six different classes within the bacterial communities. The majority were Gram-negative bacteria, with the Gammaproteobacteria class representing between 56% and 92%. The most common bacterial genera found in this study were Shewanella, Vibrio and Psychrobacter. Four species of human pathogenic bacteria were also identified: V. vulnificus, V. parahaemolyticus, V. alginolyticus, and A. hydrophila although V. vulnificus was detected only in raw oysters. PMID- 19303654 TI - Substituted imidazopyridines as potent inhibitors of HCV replication. AB - BACKGROUND/AIMS: Following lead optimization, a set of substituted imidazopyridines was identified as potent and selective inhibitors of in vitro HCV replication. The particular characteristics of one of the most potent compounds in this series (5-[[3-(4-chlorophenyl)-5-isoxazolyl]methyl]-2-(2,3 difluorophenyl)-5H-imidazo[4,5-c]pyridine or GS-327073), were studied. METHODS: Antiviral activity of GS-327073 was evaluated in HCV subgenomic replicons (genotypes 1b, 1a and 2a), in the JFH1 (genotype 2a) infectious system and against replicons resistant to various selective HCV inhibitors. Combination studies of GS-327073 with other selective HCV inhibitors were performed. RESULTS: Fifty percent effective concentrations for inhibition of HCV subgenomic 1b replicon replication ranged between 2 and 50 nM and were 100-fold higher for HCV genotype 2a virus. The 50% cytostatic concentrations were > or = 17 microM, thus resulting in selectivity indices of > or = 340. GS-327073 retained wild-type activity against HCV replicons that were resistant to either HCV protease inhibitors or several polymerase inhibitors. GS-327073, when combined with either interferon alpha, ribavirin, a nucleoside polymerase or a protease inhibitor resulted in overall additive antiviral activity. Combinations containing GS 327073 proved highly effective in clearing hepatoma cells from HCV. CONCLUSIONS: GS-327073 is a potent in vitro inhibitor of HCV replication either alone or in combination with other selective HCV inhibitors. PMID- 19303655 TI - Gilbert's syndrome and hyperbilirubinemia in protease inhibitor therapy--an extended haplotype of genetic variants increases risk in indinavir treatment. AB - BACKGROUND/AIMS: Gilbert's syndrome is a frequent genetic conjugation abnormality associated with adverse drug effects. Genetic UDP glucuronosyltransferase (UGT)1A gene variants can influence gene transcription, inducibility and glucuronidation activity. Protease inhibitors used in human immunodeficiency virus (HIV) infection and chronic viral hepatitis can inhibit UGTs. Indinavir (IDV) can lead to hyperbilirubinemia in Gilbert's syndrome (UGT1A1*28), which does not explain interindividual severity differences and may thus involve additional UGT1A variants. METHODS: One hundred and twenty-five HIV patients receiving IDV and 427 healthy blood donors were genotyped for the presence of UGT1A1*28, UGT1A3 -66T/C, UGT1A7 -57T/G, UGT1A7(N129K/R131K) using Taqman 5' nuclease assays. RESULTS: Hyperbilirubinemia was observed in 42%. UGT1A1*28 frequencies did not differ between HIV patients and controls but were significantly higher in hyperbilirubinemic patients. The frequency of homozygous carriers of the 4 UGT1A marker haplotype increased with hyperbilirubinemia affecting all patients with bilirubin levels >85 micromol/l. CONCLUSIONS: In IDV treatment the risk of severe hyperbilirubinemia is associated with genetic variants of the UGT1A3 and UGT1A7 genes in addition to Gilbert's syndrome (UGT1A1*28). This haplotype is a useful predictor of protease inhibitor-induced side effects. PMID- 19303656 TI - HDMCP uncouples yeast mitochondrial respiration and alleviates steatosis in L02 and hepG2 cells by decreasing ATP and H2O2 levels: a novel mechanism for NAFLD. AB - BACKGROUND/AIMS: To explore the uncoupling activity of hepatocelluar downregulated mitochondrial carrier protein (HDMCP) in a yeast expression system and its function in non-alcoholic fatty liver disease (NAFLD). METHODS: Molecular cloning and RT-PCR were used for yeast protein expression and uncoupling activity was assessed. Western blot analysis was used to determine HDMCP level in rat NAFLD and steatotic L02 and hepG2 cell models where their presence was confirmed by pathologic (Nile red and H-E staining) and biochemical changes. RNA interference was used to knock down HDMCP level and mitochondrial ATP and hydroperoxide levels were measured for potential mechanism exploration. RESULTS: We found a significant GDP insensitive uncoupling activity of HDMCP in yeast mitochondria and its increased expression in animal and cell models. HDMCP was significantly increased with culture time and steatosis was aggravated when HDMCP level was knocked down. Furthermore, we found that HDMCP might function through promoting ATP depletion and decreasing H(2)O(2) production. CONCLUSION: This study adds supportive data to the hypothesis that HDMCP might be a long postulated liver-specific uncoupling protein and broadens our understanding of the pathogenesis of NAFLD. More importantly, HDMCP might become a novel drug target for its ability in alleviating hepatic steatosis. PMID- 19303657 TI - Antiviral therapy for hepatitis B-related liver cancer prevention is more cost effective than cancer screening. AB - BACKGROUND/AIMS: In Australia, Asian-born populations are 6-12 times more likely to develop hepatocellular cancer (HCC) than Australian-born individuals. We therefore, modelled the consequences of different management strategies for chronic hepatitis B (CHB) in Asian-born adults aged > or = 35 years. METHODS: A Markov model compared (1) enhanced surveillance for HCC alone (HCC surveillance), or (2) enhanced HCC surveillance coupled with CHB treatment (HCC prevention) to the current practice, of low CHB treatment uptake. Patients were stratified and managed according to risk categories, based upon hepatitis B virus (HBV) viral load and alanine aminotransferase (ALT) levels. We measured costs, health outcomes [cases of HCC and deaths averted, quality-adjusted life-years (QALYs) gained] and incremental cost-effectiveness ratios (ICERs). RESULTS: HCC surveillance would cost on average AU$8479 per person, compared to AU$2632 with current clinical practice and result in a gain of 0.014 QALYs (AU$401,516/QALY gained). A HCC prevention strategy would cost on average AU$14,600 per person, result in 0.923 QALYs gained (AU$12,956/QALY gained), reduce cases of cirrhosis by 52%, HCC diagnoses by 47% and CHB-related deaths by 56%, compared to current practice. CONCLUSIONS: HCC prevention appears to be a cost-effective public health strategy in at-risk populations in Australia and is preferable to HCC surveillance as a cancer control strategy. PMID- 19303658 TI - Receptor for advanced glycation end product (RAGE)-dependent modulation of early growth response-1 in hepatic ischemia/reperfusion injury. AB - BACKGROUND/AIMS: We previously showed that blockade of RAGE significantly attenuates hepatic ischemia/reperfusion (I/R) injury in mice. Here, we identify that early growth response-1 (Egr-1) is a downstream target of RAGE in hepatic I/R injury. METHODS: Hepatic I/R was induced in male mice. Liver remnants were analyzed for induction of Egr-1 and cytokines, as well as regulation of apoptotic pathways after reperfusion. RESULTS: Egr-1 was upregulated in the liver remnants after hepatic I/R injury and was suppressed by administration of soluble RAGE or deletion of the RAGE gene. RAGE-mediated increased expression of Egr-1 upregulates a central downstream gene, MIP2. In contrast, RAGE-stimulated Egr-1 independent pathways regulate TNF-alpha production and apoptosis in response to I/R. Consistent with these findings, phospho-p44/42 and phospho-JNK MAPK and c Jun were strikingly suppressed in RAGE(-/-) versus WT mice, but not in Egr-1(-/-) mice. RAGE ligand HMGB1 was upregulated after I/R in the liver remnants. In vitro, incubation of RAGE-expressing liver dendritic cells (DCs) with recombinant HMGB-1 resulted in increased Egr-1 transcripts, in a manner suppressed by RAGE gene deletion, soluble RAGE and inhibitors of p44/p42 or JNK MAP kinase. CONCLUSIONS: Suppression of Egr-1 may contribute to the protective mechanisms underlying the beneficial impact of RAGE blockade or deletion. PMID- 19303659 TI - Validation of a three-dimensional facial scanning system based on structured light techniques. AB - The aim of this study was to validate a newly developed three-dimensional (3D) structured light scanning system in recording the facial morphology. The validation was performed in three aspects including accuracy, precision and reliability. The accuracy and precision were investigated using a plaster model with 19 marked landmarks. The accuracy was determined by comparing the coordinates from the 3D images and from the coordinates measure machine (CMM). The precision was quantified through the repeated landmarks location on 3D images. The reliability was investigated in 10 adult volunteers. Each was scanned five times in 3 weeks. The 3D images acquired at different times were compared with each other to measure the reliability. We found that the accuracy was 0.93 mm, the precision was 0.79 mm, the reliability was 0.2mm. These findings suggested that the structured light scanning system was accurate, precise and reliable to record the facial morphology for both clinic and research purposes. PMID- 19303660 TI - HANOIPC3: a computer program to evaluate executive functions. AB - This article describes a computer program (HANOIPC3) based on the Tower of Hanoi game that, by analyzing a series of parameters during execution, allows a fast and accurate evaluation of data related to certain executive functions, especially planning, organizing and problem-solving. This computerized version has only one level of difficulty based on the use of 3 disks, but it stipulates an additional rule: only one disk may be moved at a time, and only to an adjacent peg (i.e., no peg can be skipped over). In the original version--without this stipulation--the minimum number of movements required to complete the task is 7, but under the conditions of this computerized version this increases to 26. HANOIPC3 has three important advantages: (1) it allows a researcher or clinician to modify the rules by adding or removing certain conditions, thus augmenting the utility and flexibility in test execution and the interpretation of results; (2) it allows to provide on-line feedback to subjects about their execution; and, (3) it creates a specific file to store the scores that correspond to the parameters obtained during trials. The parameters that can be measured include: latencies (time taken for each movement, measured in seconds), total test time, total number of movements, and the number of correct and incorrect movements. The efficacy and adaptability of this program has been confirmed. PMID- 19303661 TI - Endogenous bone morphogenetic proteins in human bone marrow-derived multipotent mesenchymal stromal cells. AB - Primary human multipotent mesenchymal stromal cells (MSCs) are capable of self renewal or differentiation into several different lineages, including osteoblasts, chondrocytes and adipocytes. However, upon prolonged in vitro culture, MSCs tend to undergo spontaneous osteogenic differentiation. Here, we address the possible role of endogenous osteogenic bone morphogenetic proteins (BMPs) in in situ osteoblastic differentiation of human MSCs. Human MSCs consistently express biologically active BMP-2, BMP-4 and BMP-6 in addition to all BMP-activated receptors, which are functional as shown by the induction of alkaline phosphatase (ALP) activity and up-regulation of osteogenic genes (ALP, BSP1, collagen I and Runx2) following BMP-2 exposure. Since glycosaminoglycans (GAGs) have been implicated in the modulation of the osteogenic bioactivity of BMPs, we reduced sulphated cell surface GAGs by NaClO(3) treatment and found significantly reduced osteogenic gene expression and ALP activity, suggesting that this was partly due to the reduced biological activity of endogenous BMPs. Antagonising osteogenic BMP activity led to a significant reduction in the ALP activity and down-regulation of the transcription factor Runx2 associated with osteogenic development. Blocking BMP receptor type I kinase function with dorsomorphin demonstrated that endogenous osteogenesis was independent of Smad activation but was dependent on phosphatidylinositol 3-kinase (PI-3K). Inclusion of the PI-3K kinase inhibitor Ly294002 significantly reduced osteogenic gene expression and ALP activity. Spontaneous mineralisation was also abrogated following PI-3K inhibition. Thus, endogenous BMPs could contribute to spontaneous osteogenesis through Smad-independent PI-3K-dependent signalling. PMID- 19303662 TI - Candida colonisation as a source for candidaemia. AB - Candida spp. are important healthcare-associated pathogens. Identifying the source of infection is important for prevention and control strategies. The objective of this study was to evaluate candida colonisation sites as potential sources for candidaemia. Sixty-three consecutive patients with a positive blood culture for candida were included. Surveillance cultures were collected from urine, rectum, oropharynx, skin, intravascular catheter tip and skin around catheter. Molecular typing was performed when the same species of candida was isolated from blood and surveillance sites of a patient. C. albicans was associated with 42% of candidaemias, C. parapsilosis 33%, C. tropicalis 16% and C. guilliermondii, C. krusei, C. glabrata, C. holmii and C. metapsilosis were all 2% each. Six of 10 C. parapsilosis catheter tip isolates were indistinguishable from corresponding blood isolates (all in neonates). C. albicans isolates from blood were indistinguishable from corresponding gastrointestinal tract isolates in 13 of 26 patients and from catheter tip isolates in two patients. In conclusion, the results suggest that gastrointestinal colonisation is the probable source of C. albicans candidaemia and C. parapsilosis is exogenous. PMID- 19303663 TI - Identification of nigral dopaminergic neuron-enriched genes in adult rats. AB - Dopaminergic (DA) neurons in the substantia nigra play crucial roles in movement control and other physiological activities. Degeneration of these neurons is closely associated with Parkinson's disease. However, the molecular identity of nigral DA neurons is not fully understood. To identify nigral DA neuron-enriched genes, we used microarrays to compare the genome-wide gene expression profiles in 6-hydroxydopamine-lesioned, and control, substantia nigra. We identified a total of 88 unique differentially expressed gene transcripts. The spatial expression patterns of a set of these genes, including Slc10a4, Rit2, F2r, Snx10 and Slc24a3, were validated by in situ hybridization. It was revealed that their expression was highly specific in the substantia nigra. Thus we identified a set of genes that are highly expressed in nigral DA neurons, and may be involved in the maintenance and survival of nigral DA neurons in the adult rat brain. Our study also provides a general approach for profiling cell type-specific gene expression in the mature mammalian brain. PMID- 19303664 TI - [Use of psychoactive drugs and family dysfunction]. AB - OBJECTIVE: To find out the relationship between the use of psychoactive drugs in women and the results of the APGAR and stressful life events (SLE) questionnaires. DESIGN: Cross-sectional, descriptive study. SETTING: General Fanjul Health Centre in Area 7 of the Community of Madrid. PARTICIPANTS: All the women between 25-65 years who were seen in Primary Care clinics in one week in March, 2005. METHOD: Validated surveys validated of family dysfunction (Apgar) and stressful life events (additive scale of evaluation of social readjustment) were carried on women between 25-65 years who took psychoactive drugs. RESULTS: Of the 625 women who were seen in the clinics, 121 (19.36%), with a mean age of 46.8+/-12.7 years, were interviewed. Of these, 90% were Spanish, 55.4% were married, 67% had children, 3.3% had not received schooling, 75.2% had a primary school education, 21.5% with university studies, and 57.9% work outside the home. Antidepressants were used by 24% of the women, with 30.6% taking benzodiazepines. The use of benzodiazepines increases with the age (p<0.001). On having look at the relationship between the use of benzodiazepines or antidepressants and the existence of major family dysfunction or major stressful life events (SLE) we verify that statistically significant differences do not exist. CONCLUSIONS: Although it was intuitively possible to believe that SLE and family dysfunction lead to a higher use of psychoactive drugs, we could not find a statistically significant relationship. We have shown that there is a higher use of benzodiazepines in older women. PMID- 19303665 TI - [How do we manage information in the clinic to guarantee quality health care]. PMID- 19303666 TI - [Cooperation between primary care and mental health services]. AB - PURPOSE: To find out the current coordination/cooperation level of primary care (PC) with the adult mental health centres (AMHC), the addiction treatment centres (ATC) and children and adolescent mental health centres (CAMHC) of Catalonia (Spain). Differences in coordination between urban and non-urban areas were also evaluated. DESIGN: Cross-sectional descriptive study. SETTING: Eighty PC centres representing the seven health regions of Catalonia. PARTICIPANTS: Selection was by means of a multi-staged and stratified sampling method. A total of 356 of the 618 PC physicians who agreed to participate completed a survey evaluating different aspects of coordination with the specialised mental health services. RESULTS: The PC physicians indicated that the availability of AMHC was worse than that provided by ACT and CAMHC. However, the reports on the state of the referred patients and the training courses in mental health offered by the AMHC were more frequent. There were no significant differences in availability or training in mental health between urban and non-urban areas. CONCLUSIONS: The cooperation between the PC and the specialised mental health services of Catalonia is optimal in some aspects, but patient waiting time needs to be reduced for first visits to the AMHC and training activities offered by the CAMHC and the ACT need to be increased. PMID- 19303667 TI - [Coordination with mental health services. A pending subject, manifestly improvable]. PMID- 19303668 TI - [Aminoglycosides and polymyxins]. AB - The emergence of multidrug-resistant strains of gram-negative bacilli during the last decade has generated renewed interest in older antimicrobials that had been relegated to a second line because of a poorer safety profile, but that are still active against these microorganisms. Once-daily administration of aminoglycosides has limited the toxicity of these agents and enabled their reintroduction into clinical practice. Recent studies have shown no additional benefits of concomitant administration of aminoglycosides with current ss-lactams, and the available evidence does not support the use of once-daily administration for all indications. The new formulations and dosages of polymyxins have also reduced the toxicity rates attributed to these agents in the past. Although more extensive studies are required to properly define their pharmacokinetics and effectiveness, the available data have shown favorable outcomes for patients with infection due to multiresistant gram-negative bacilli treated with colistin, either alone or combined with other antimicrobial agents. PMID- 19303669 TI - [Case-crossover design: Basic essentials and applications]. AB - Case-crossover analysis is an observational epidemiological design that was proposed by Maclure in 1991 to assess whether a given intermittent or unusual exposure may have triggered an immediate short-term, acute event. The present article outlines the basics of case-crossover designs, as well as their applications and limitations. The case-crossover design is based on exclusively selecting case subjects. To calculate relative risk, exposure during the period of time prior to the event (case period) is compared against the same subject's exposure during one or more control periods. This method is only appropriate when the exposures are transient in time and have acute short-term effects. For exposures in which there is no trend, a unidirectional approach is the most frequent and consists of selecting one or more control periods prior to the case period. When the exposure displays a time trend (e.g., air pollution), a unidirectional approach will yield biased estimates, and therefore bidirectional case-crossover designs are used, which select control time intervals preceding and subsequent to that of the event. The case-crossover design is being increasingly used across a wide range of fields, including factors triggering traffic, occupational and domestic accidents and acute myocardial infarction, and those involved in air pollution and health and pharmacoepidemiology, among others. Insofar as data-analysis is concerned, case-crossover designs can generally be regarded as matched case-control studies and consequently conditional logistic regression can be applied. Lastly, this study analyzes practical examples of distinct applications of the case-crossover design. PMID- 19303670 TI - Identification of putative steroid-binding sites in human ABCB1 and ABCG2. AB - Homology modelling was used to generate three-dimensional structures of the nucleotide-binding domains (NBDs) of human ABCB1 and ABCG2. Interactions between a series of steroidal ligands and transporter NBDs were investigated using an in silico docking approach. C-terminal ABCB1 NBD (ABCB1 NBD2) was predicted to bind steroids within a cavity formed partly by the P-Loop, Tyr1044 and Ile1050. The P Loop within ABCG2 NBD was also predicted to be involved in steroid binding. No overlap between ATP- and RU-486-binding sites was predicted in either NBD, though overlaps between ATP- and steroid-binding sites were predicted in the vicinity of the P-Loop in both nucleotide-binding domains. PMID- 19303672 TI - [A cardiorespiratory decompensation in the course of rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) may include rare extra-articular manifestations, and particularly cardiopulmonary involvement. We report a 83-year-old woman, with RA, who presented with both pulmonary and heart failures that were related to the RA, resulting from the presence of mediastinal and axillary lymph nodes, and pleural and pericardial effusions. This was confirmed by the dramatic improvement after a specific treatment of RA. PMID- 19303673 TI - [Osteonecrosis of the jaw and bisphosphonates: current data and perspectives]. PMID- 19303674 TI - Can interprofessional education DVD simulations provide an alternative method for clinical placements in nursing? AB - PURPOSE: The study aimed to evaluate the usability of DVD simulations, the impact on student learning, clinical placement orientation, and the potential for using DVD simulations to reduce the clinical placement burden on the health care system with nursing students. METHOD: A total of 11 DVD simulations were developed by Monash University academics. Second year students (N=191) from the Bachelor of Nursing course at Monash University, viewed a range of DVDs. Students' perceptions and attitudes about the clinical relevance of the simulations were assessed by having them complete a 7-point Likert self-report scale. Qualitative data was also collected from two focus groups (N=7). RESULTS: Overall, nursing students perceived the DVD simulations positively in relation to learning attention (M=4.93, SD=1.02, CI 4.25-4.54), learning potential (M=4.45, SD= 1.30, CI 5.13-5.50), clinical relevance to practice (M=5.32, SD=0.65, CI 4.36-4.55), and information processing quality (M=5.62, SD= 1.02, CI 5.47-5.76). The following themes emerged from the focus groups: provided familiarisation for clinical placements, learning wastage occurs in varying amounts, simulations could replace some clinical placement rotations, supportive of multidisciplinary approach and integration, and simulations should have pedagogical integration into weekly clinical cases. CONCLUSION: Nursing students reported that the simulations were educationally, professionally, and clinically relevant. The cost benefit of using DVD simulations as an alternative and potential replacement to elements of nursing clinical placements should be investigated further. PMID- 19303675 TI - Short- and mid-term effects of a setting based prevention program to reduce obesity risk factors in children: a cluster-randomized trial. AB - BACKGROUND & AIMS: To assess the effects of a low cost behavioral prevention program in a preschool setting. METHODS: 64 Kindergartens in 4 Bavarian regions were randomly assigned as intervention or controls in a 2:1 ratio. Samples of 1318 and 1340 children examined in the school entrance health examination at 5.7+/-2.6 and 17.6+/-2.3 months (mean+/-standard deviation for first and second sample) after the start of the program were analysed. MEASUREMENTS: Main outcome measures were the prevalence of high fruit and vegetable consumption, low consumption of high caloric drinks assessed in parental questionnaires, overweight and obesity, and secondary, further dietary habits and results of motoric testing. RESULTS: The program led to an increased proportion of children with high fruit and vegetable consumption already after 6 months, which was sustainable with adjusted odds ratios of 1.59 (1.26: 2.01) and 1.48 (1.08: 2.03) after 18 months. Subgroup analyses by gender, overweight and parental education, performed in order to assess consistency of effects, showed similar results. Prevalence of overweight and obesity as well as motoric testing results were not statistically different between intervention and control groups. CONCLUSION: This low cost setting based behavioral intervention achieved sustainable effects on fruit and vegetable consumption in young children 18 months after the start of the intervention and showed effects also in the high risk groups of children from families with lower education levels, and children already overweight. PMID- 19303676 TI - The changing face of oral and maxillofacial surgery. PMID- 19303677 TI - Mothers' views of caseload midwifery and the value of continuity of care at an Australian regional hospital. AB - OBJECTIVE: to evaluate mothers' satisfaction with a caseload-midwifery scheme, and whether this varied according to the extent of continuity of care provided. DESIGN: mothers' satisfaction with care was assessed using a postal survey, which was linked with their clinical outcomes data. SETTING: the Wollongong Hospital, the major public hospital in the Illawarra region, New South Wales, Australia. PARTICIPANTS: 174 women gave birth during the evaluation period of a pilot midwifery group practice (MGP) programme at the Wollongong Hospital between 5 July 2004 and 30 April 2005. This group included 87 primiparous and 87 multiparous women, all of whom met the Australian College of Midwives' criteria for low-risk pregnancies. INTERVENTION: the MGP was staffed by six midwives working in two groups of three. Each midwife took on a primary caseload of 40 women per year, and provided support as a secondary midwife to women cared for by colleagues. MEASUREMENTS: mothers' satisfaction with care and adjustment to motherhood were assessed with self-completed questionnaires. Survey responses were linked with clinical data, allowing examination of the relationship between maternal satisfaction and continuity of care. FINDINGS: the MGP achieved high levels of continuity of care, both objectively (based on birth records) and from mothers' perspectives. Overall, mothers' evaluations of their care were very positive. Women indicated that their relationships with their midwives were genuinely caring and a valued source of reassurance and comfort during pregnancy, labour and early motherhood. Although continuity of care did not predict summary scores for maternal satisfaction, it was related to some individual items on the satisfaction scales. Satisfaction with control and communication was predicted by parity and the level of intervention during labour and birth. KEY CONCLUSIONS: continuous care appears to facilitate the development of supportive relationships between women and their midwives. Women's perceptions about continuous and respectful treatment were related to objectively measured continuity of care. The qualitative data confirm the importance of less tangible benefits, such as the quality of relationships between women and their caregivers. IMPLICATIONS FOR PRACTICE: the viability of caseload-midwifery-led care for low-risk pregnancies depends, in part, on the model's acceptability to consumers. This study demonstrated that the caseload model is associated with high levels of maternal satisfaction. Supportive relationships with midwives in a caseload scheme are highly valued by women. PMID- 19303678 TI - Association between micronuclei frequency in pollen mother cells of Tradescantia and mortality due to cancer and cardiovascular diseases: a preliminary study in Sao Jose dos Campos, Brazil. AB - The present study was designed to explore the correlation between the frequency of micronuclei in Trad-MN, measured across 28 biomonitoring stations during the period comprised between 11 of May and 2 of October, 2006, and adjusted mortality rates due to cardiovascular, respiratory diseases and cancer in Sao Jose dos Campos, Brazil, an area with different sources of air pollution. For controlling purposes, mortality rate due to gastrointestinal diseases (an event less prone to be affected by air pollution) was also considered in the analysis. Spatial distribution of micronuclei frequency was determined using average interpolation. The association between health estimators and micronuclei frequency was determined by measures of Pearson's correlation. Higher frequencies of micronuclei were detected in areas with high traffic and close to a petrochemical pole. Significant associations were detected between micronuclei frequency and adjusted mortality rate due to cardiovascular diseases (r=0.841, p=0.036) and cancer (r=0.890, p=0.018). The association between mortality due to chronic obstructive pulmonary diseases was positive but did not reach statistical significance (r=0.640, p=0.172), probably because of the small number of events. Gastrointestinal mortality did not exhibit significant association with micronuclei frequency. Because the small number of observations and the nature of an ecological study, the present findings must be considered with caution and considered as preliminary. Further studies, performed in different conditions of contamination and climate should be done before considering Trad-MN in the evaluation of human health risk imposed by air pollutants. PMID- 19303679 TI - Rapamycin for focal segmental glomerulosclerosis: a report of 3 cases. AB - Corticosteroids and/or cyclosporine constitute the present therapeutic approach for patients with focal segmental glomerulosclerosis (FSGS). The high incidence of side effects for the former and risk of nephrotoxicity combined with the high relapse rate after discontinuation for the latter render their use problematic. Results concerning the role of rapamycin in the treatment of patients with FSGS are conflicting. We describe results for 3 patients treated with a combination of low-dose steroids and rapamycin for FSGS, focusing on the importance of maintaining low drug (rapamycin) levels by using a twice-daily regimen. PMID- 19303680 TI - Critical care nephrology: Core Curriculum 2009. PMID- 19303681 TI - Lethal cystic kidney disease in Amish neonates associated with homozygous nonsense mutation of NPHP3. AB - BACKGROUND: Nephronophthisis is a group of genetically heterogeneous autosomal recessive cystic kidney disorders with a wide spectrum of severity and age of onset. We present a clinical and genetic study of a lethal form of nephronophthisis in neonates. STUDY DESIGN: Clinical and genetic investigations of a case series. SETTING & PARTICIPANTS: 12 affected offspring born to consanguineous parents from the Old Order Amish community. OUTCOMES: In this extended pedigree, the disorder is particularly severe; affected individuals survive only hours or days, with the cause of death invariably respiratory distress. RESULTS: Cystic kidneys were confirmed in 11 infants and suspected in an additional individual who had 2 affected siblings. Although the renal aspect of the phenotype was a consistent feature in all affected individuals, additional pulmonary, cardiac, and urinary tract abnormalities are variable parts of this syndrome. Physical mapping of the causative mutation in this extended Amish pedigree highlighted a 475-kilobase candidate region on chromosome 3 that contains the NPHP3 gene. Sequence analysis of this gene showed a cytosine to thymine substitution in exon 15 (c.2104C-->T) that cosegregated with the disease status. This substitution is predicted to lead to premature termination at position 702 of the protein product (p.Arg702X). LIMITATIONS: Because of the severe nature of this disease, few affected infants underwent full clinical evaluation. CONCLUSION: The presence of congenital malformations in the case series confirms the crucial role of NPHP3 in early embryonic development of the kidneys and urinary tract. The study also highlights the subtle variations in phenotypic expression in a cohort of patients with the same mutation in NPHP3. PMID- 19303682 TI - Therapeutic implications of coexisting severe pulmonary hemorrhage and pulmonary emboli in a case of Wegener granulomatosis. AB - Wegener granulomatosis classically involves the renal, respiratory, and ear, nose, and throat systems. Pulmonary hemorrhage is recognized as a severe respiratory complication. Untreated, the mortality rate approaches 90% at 2 years. We describe a case of Wegener granulomatosis with coexistent severe lung hemorrhage and pulmonary and deep vein thromboses. A 31-year-old man presented with features of vasculitis, including epistaxis, fever, and acute kidney injury with an increased serum creatinine level (3.27 mg/dL). Kidney biopsy confirmed pauci-immune crescentic glomerulonephritis, and antineutrophil cytoplasmic antibody showing a cytoplasmic staining pattern was strongly positive. Standard immunosuppression therapy (prednisolone and cyclophosphamide) was started. Eleven days later, the patient developed sudden dyspnea. A computed tomographic pulmonary angiogram showed pulmonary emboli, and ultrasound of the limbs showed ileofemoral thrombi bilaterally. Subcutaneous enoxaparin and warfarin therapy was started, but 8 days later, the patient had a massive pulmonary hemorrhage. Anticoagulation therapy was stopped, and plasma exchange was started to prevent further life-threatening hemorrhage. An inferior vena cava filter was inserted to prevent further pulmonary emboli during the period when anticoagulation was withheld. Kidney function improved, and pulmonary hemorrhage resolved after 5 plasma exchanges. Reintroduction of intravenous heparin and subsequently warfarin caused no further bleeding. We discuss the difficult management dilemma this combination of disease manifestations presents and review the current literature. PMID- 19303684 TI - Long-term outcomes after acute kidney injury: where we stand and how we can move forward. PMID- 19303683 TI - Uric acid and long-term outcomes in CKD. AB - BACKGROUND: Hyperuricemia is prevalent in patients with chronic kidney disease (CKD); however, data are limited about the relationship of uric acid levels with long-term outcomes in this patient population. STUDY DESIGN: Cohort study. SETTING & PARTICIPANTS: The Modification of Diet in Renal Disease (MDRD) Study was a randomized controlled trial (N = 840) conducted from 1989 to 1993 to examine the effects of strict blood pressure control and dietary protein restriction on progression of stages 3 to 4 CKD. This analysis included 838 patients. PREDICTOR: Uric acid level. OUTCOMES & MEASUREMENTS: The study evaluated the association of baseline uric acid levels with all-cause mortality, cardiovascular disease (CVD) mortality, and kidney failure. RESULTS: Mean age was 52 +/- 12 (SD) years, glomerular filtration rate was 33 +/- 12 mL/min/1.73 m(2), and uric acid level was 7.63 +/- 1.66 mg/dL. During a median follow-up of 10 years, 208 (25%) participants died of any cause, 127 (15%) died of CVD, and 553 (66%) reached kidney failure. In multivariate models, the highest tertile of uric acid was associated with increased risk of all-cause mortality (hazard ratio [HR], 1.57; 95% confidence interval [CI], 1.07 to 2.32), a trend toward CVD mortality (HR, 1.47; 95% CI, 0.90 to 2.39), and no association with kidney failure (HR, 1.20; 95% CI, 0.95 to 1.51) compared with the lowest tertile. In continuous analyses, a 1-mg/dL greater uric acid level was associated with 17% increased risk of all-cause mortality (HR, 1.17; 95% CI, 1.05 to 1.30) and 16% increased risk of CVD mortality (HR, 1.16; 95% CI, 1.01 to 1.33), but was not associated with kidney failure (HR, 1.02; 95% CI, 0.97 to 1.07). LIMITATIONS: Primary analyses were based on a single measurement of uric acid. Results are generalizable primarily to relatively young white patients with predominantly nondiabetic CKD. CONCLUSIONS: In patients with stages 3 to 4 CKD, hyperuricemia appears to be an independent risk factor for all-cause and CVD mortality, but not kidney failure. PMID- 19303685 TI - Thrombocytosis in EPO-treated dialysis patients may be mediated by EPO rather than iron deficiency. PMID- 19303686 TI - Cryoglobulinemia and glomerular rhomboid inclusions in a child with acute kidney injury. AB - Cryoglobulinemia is rarely reported in children, and kidney failure secondary to cryoglobulinemia is even more uncommon. We report the case of a 7-year-old boy with cryoglobulins and a systemic illness, including persistent fever, arthralgias, rash, hypocomplementemia, and acute kidney injury associated with nephritic urine sediment. An extensive workup showed no infectious, neoplastic, or rheumatological cause of his kidney injury. The kidney biopsy specimen showed membranoproliferative glomerulonephritis type 1 with electron microscopic evidence of rhomboid crystalloid inclusions. These inclusions have rarely been reported in adult patients with cryoglobulinemia. The patient underwent spontaneous remission, including full recovery of kidney function, and required no immune suppression. The patient's course is consistent with cryoglobulinemia associated kidney injury, which supports the inclusion of essential cryoglobulinemia in the differential diagnosis of pediatric patients with hypocomplementemic glomerulonephritis. PMID- 19303687 TI - The independent contribution of individual-, neighbourhood-, and country-level socioeconomic position on attitudes towards intimate partner violence against women in sub-Saharan Africa: a multilevel model of direct and moderating effects. AB - We examined associations between country, neighbourhood, and individual socioeconomic position (SEP) and attitudes towards intimate partner violence against women (IPVAW). We applied multivariable multilevel logistic regression analysis on Demographic and Health Survey data for 165,983 women and 68,501 men nested within 7465 communities from 17 countries in sub-Saharan Africa collected between 2003 and 2007. Contrary to expectation women were 34% more likely to justify IPVAW than men. We found that sex moderates associations of individual-, neighbourhood-, and country-level SEP with attitudes towards IPVAW. There was a significant positive interaction effect between sex and education attainment; women with no education were more likely to justify IPVAW than men with no education. Negative sex interaction with household wealth status indicates that differences in attitude are less pronounced among women. Unemployed men were more likely to justify IPVAW. Interaction effects indicate that the association of neighbourhood socioeconomic disadvantage with attitudes was more pronounced among women than among men. The association of country-level SEP with attitudes towards IPVWA was inconclusive. There was some evidence that neighbourhood modified the association between individual SEP and attitudes towards IPV. Also, there was cross-level interaction between country and neighbourhood SEP. Neighbourhood and individual SEP were independently associated with attitudes towards IPVAW. The relationship with country-level SEP was inconclusive. The findings underscore the need to implement public health prevention/intervention strategies not only at the level of individual SEP but also at the neighbourhood level. PMID- 19303688 TI - Is education the link between orphanhood and HIV/HSV-2 risk among female adolescents in urban Zimbabwe? AB - As the population of orphans grows in AIDS-affected settings, recent studies describe a heightened risk of HIV and sexual risk behaviours among adolescent orphans compared to their non-orphaned peers. This study explores the role of education in explaining the excess sexual risk previously documented among unmarried female orphans in urban Zimbabwe. School attendance and attainment were assessed by type of orphanhood, and for their association with markers of sexual risk (HIV and/or HSV-2 infection) among 743 participants drawn from a random sample of 15-19-year-old girls identified in a cross-sectional survey in Highfield, Harare, in 2004. Multivariable logistic regression was used to assess the role of educational status in explaining the higher prevalence of adverse sexual outcomes among unmarried orphans compared to non-orphans, adjusting for possible confounders. Double orphans had significantly lower educational attendance and attainment than non-orphans. Maternal orphans had higher odds of school drop-out, although this association disappeared when adjusted for recent mobility. Educational status was strongly associated with HIV/HSV-2 risk, but explained only a small part of double orphans' sexual risk and did not explain the HIV/HSV-2 risk of maternal and paternal orphans. High overall levels of secondary school participation and school fee assistance provided to vulnerable families may have reduced the schooling disparities between orphans and non orphans in Highfield. However, further efforts are needed to rectify the schooling inequities that persist, while additional research is needed to identify other socio-economic and emotional factors driving orphans' sexual risk so that prevention and support programmes can meet the needs of this growing population. PMID- 19303689 TI - How Cuba's Latin American School of Medicine challenges the ethics of physician migration. AB - This paper demonstrates a working alternative to the accepted ethics of physician migration. A dominant cosmopolitan ethics encourages upward mobility of physicians in a globalized labour force, and this ultimately advances the position of individuals rather than improving public health-care service for vulnerable communities in the global South. Cuba's Escuela Latinoamericana de Medicina (ELAM) challenges this trend as its institutional ethics furnishes graduates with appropriate skills, knowledge and service ethics to deliver quality care in marginalized areas. This paper provides an analysis of how ELAM trains physicians in community-oriented service for marginalized areas in the global South. The principle finding of this analysis is that ELAM exhibits a working alternative to the accepted ethics of physician migration, as it encourages graduates to practice in marginalized communities rather than feed the migration pipeline into the North. Arguably, ELAM serves as an important case study in how a medical school's ethics can work to bring graduates closer to the communities that are in desperate need of their skills and of their compassion. PMID- 19303690 TI - Dynamics of female pelvic floor function using urodynamics, ultrasound and Magnetic Resonance Imaging (MRI). AB - In this review the diagnostic potential of evaluating female pelvic floor muscle (PFM) function using magnetic and ultrasound imaging in the context of urodynamic observations is considered in terms of determining the mechanisms of urinary continence. A new approach is used to consider the dynamics of PFM activity by introducing new parameters derived from imaging. Novel image-processing techniques are applied to illustrate the static anatomy and dynamics of PFM function of stress incontinent women pre- and post-operatively as compared to asymptomatic subjects. Function was evaluated from the dynamics of organ displacement produced during voluntary and reflex activation. Technical innovations include the use of ultrasound analysis for movement of structures during maneuvers that are associated with external stimuli. Enabling this approach is the development of criteria and fresh and unique parameters that define the kinematics of PFM function. Principal among these parameters, are displacement, velocity, acceleration and the trajectory of pelvic floor landmarks. To accomplish this objective, movement detection, including motion tracking algorithms and segmentation algorithms were developed to derive new parameters of trajectory, displacement, velocity and acceleration, and strain of pelvic structures during different maneuvers. Results highlight the importance of timing the movement and deformation to fast and stressful maneuvers, which are important for understanding the neuromuscular control and function of PFM. Furthermore, observations suggest that timing of responses is a significant factor separating the continent from the incontinent subjects. PMID- 19303691 TI - The role of ultrasound in assessment of male fertility. AB - Ultrasonography (US) is a widely used and well tolerated imaging modality for evaluation of pathologic conditions of the testes. Recent technical advances of US applications and post processing developments have enabled new aspects in the structural and functional analysis of testicular tissue and therefore male fertility. This review covers the most relevant approaches due to recent technical advances. Testicular volume measured by B-mode US correlated significantly with testicular function. Increased resistive index (RI) and pulsatility index (PI) of capsular branches of testicular arteries on unenhanced color Doppler US examination may be an indicator of impaired testicular microcirculation in patients with clinical varicocele. FSH was inversely correlated with testicular volume and directly correlated with testicular vascularization, suggesting that ultrasonographic and color Doppler scanning of the testes may be used, if a sperm count is not available, to indirectly assess the gonadal function. Perfusion mapping, performed with the use of color Doppler ultrasound, has shown for the first time that in patients suffering from azoospermia, sperm quality and quantity depend on tissue perfusion within the testicle. Testicular arterial blood flow was found to be significantly decreased in men with varicocele. This may be a reflection of the impaired microcirculation. Following decreased testicular arterial blood flow, impaired spermatogenesis may result from defective energy metabolism in the microcirculatory bed. Contrast-enhanced ultrasound (CEUS) imaging is potentially applicable to the investigation of vascular disorders of the testis. Pulse inversion (PI) US data can correctly determine relative testicular perfusion based on nonlinear curve fitting of the US backscatter intensity as a function of time and spectral analysis of the intensity time trace. PI imaging, compared with conventional Doppler US methods, provides superior assessment of perfusion in the setting of acute testicular ischemia. New contrast-enhanced US techniques like microvessel imaging (proposed by Philips) and CPS (proposed by Siemens-Acuson) allow for a better determination of tissue perfusion based on time intensity curves and an illustration of vessel distribution inside the testis. First results show a lower vessel density in atrophic testes and a difference in contrast dynamics in testes with impaired function. Real-time elastography, a method for illustration of tissue stiffness under real-time conditions, demonstrates different elasticity values dependent on testicular volume and function. Further studies will prove if these techniques can evolve into clinical practice. PMID- 19303692 TI - Cervical ripening and insufficiency: from biochemical and molecular studies to in vivo clinical examination. AB - To understand cervical ripening and especially the pathophysiology of cervical insufficiency, it is important to know the cervical composition: the cervix is dominated by fibrous connective tissue, consisting predominantly of Type I collagen (70%). Despite many studies of the cervix, we still rely upon relatively crude methods for clinical evaluation of the cervix. If the amount of cervical collagen plays a role in cervical insufficiency and in success of or length of induction of labor, then measurements of cervical collagen may provide an objective means of establishing the diagnosis or prognosis. We have established and reported a non-invasive means, called Collascope, to measure collagen cross linking using light-induced fluorescence (LIF), and which is specifically designed to assess cervical ripening, and functions by measuring the natural fluorescence of non-soluble collagen in the cervix. Studies conducted in animals and humans in a variety of settings indicate that cervical function can be successfully monitored using the Collascope during pregnancy: LIF correlates negatively with gestational age and positively with time-to-delivery interval, and is predictive of delivery within 24h. Additionally LIF is significantly lower in women with cervical insufficiency. We suggest that the Collascope might be useful to better define management in cases of spontaneous preterm or induced term cervical ripening. From our studies and others, it is clear that in forecasting (pre-)term cervical ripening, the capability of the technologies and bioassays that have been generally accepted into clinical practice are limited. Any devices shown to be superior to the clinically accepted tests currently used should be quite useful for clinicians. The Collascope offers an objective measurement of both the function and state of the cervix, by directly measuring collagen cross-linking using LIF. PMID- 19303693 TI - Changes in the biochemical constituents and morphologic appearance of the human cervical stroma during pregnancy. AB - OBJECTIVE: The cervix is the lower portion of the uterus. It is composed of fibrous tissue and its mechanical integrity is crucial for maintaining a healthy gestation. During normal pregnancy, the cervical extracellular matrix (ECM) remodels in preparation for labor. The objective of this study was to investigate the biochemical and morphological changes in cervical stroma associated with physiological remodeling during normal pregnancy. STUDY DESIGN: Using human cervical tissue obtained from pregnant and non-pregnant patients, the ECM was analyzed for its biochemical constituents and histologic morphology. The ECM was assayed for hydration, collagen concentration, collagen solubility, total sulfated glycosaminoglycan concentration, and individual disaccharide concentration. The ECM morphology was visualized using conventional histological techniques (Masson's trichrome stain, polarized light microscopy) as well as second harmonic generation (SHG) imaging. RESULTS: When comparing pregnant to non pregnant tissue, significant increases were measured for total sulfated glycosaminoglycans, hyaluronic acid, and collagen solubility. The microscopy studies confirmed that the collagenous network of the cervical stroma was anisotropic and pregnancy was associated with a discernable decrease in collagen organization. CONCLUSION: Significant changes were seen in the concentration and organization of cervical ECM constituents during normal pregnancy. PMID- 19303694 TI - Transmission line models to simulate the impedance of the uterine vasculature during the ovarian cycle and pregnancy. AB - OBJECTIVES: Changes in uterine vascular impedance may yield diagnostic insight into physiological and pathological changes in uterine vascular resistance and compliance during the ovarian cycle and pregnancy. Herein, our objectives were to develop models to simulate uterine vascular impedance in order to gain insight into the vascular size and stiffness changes that occur during ovarian cycling and pregnancy. STUDY DESIGN: Two electrical analogue transmission line models were developed and evaluated based on goodness-of-fit to experimental impedance measurements, which were obtained in nonpregnant luteal and follicular phase (NP L and NP-F) and pregnant (P) ewes (n=4-8 per group). First, an anatomically based, multi-segment, symmetric, branching transmission line model was developed. Parameter values were calculated based on experimental measurements of size and stiffness in the first three generations of the uterine arterial tree for NP-L, NP-F and P ewes. Then, a single segment transmission line model was developed and effective parameter values were optimized to best-fit the measured impedances. RESULTS: The anatomically based multi-segment model did not yield the expected good agreement with the experimental data (R(2)<0.5 for all groups). In contrast, the impedance spectra predicted by the single segment model agreed very well with experimental data (R(2)=0.93, 0.82, and 0.84 for NP-L, NP-F and P, respectively; p<0.0001, all groups). Furthermore, the changes in the best-fit model parameters for NP-F and P compared to the NP-L were consistent with the prior literature on the effects of the ovarian cycle and pregnancy on vascular resistance and compliance. In particular, compared to NP-L, NP-F had decreased longitudinal and terminal resistance with a modest increase in compliance whereas pregnancy caused more dramatic drops in longitudinal and terminal resistance and a significant increase in compliance. CONCLUSIONS: The single segment transmission line model is a useful tool to examine changes in vascular structure and function that occur during the ovarian cycle and pregnancy. PMID- 19303695 TI - Development of a new method for monitoring prostate-specific antigen changes in men with localised prostate cancer: a comparison of observational cohorts. AB - BACKGROUND: Prostate-specific antigen (PSA) measurements are increasingly used to monitor men with localised prostate cancer (PCa), but there is little consensus about the method to use. OBJECTIVE: To apply age-specific predictions of PSA level (developed in men without cancer) to one cohort of men with clinically identified PCa and one cohort of men with PSA-detected PCa. We hypothesise that among men with clinically identified cancer, the annual increase in PSA level would be steeper than in men with PSA-detected cancer. DESIGN, SETTING, AND PARTICIPANTS: The Scandinavian Prostate Cancer Group 4 (SPCG-4) cohort consisted of 321 men assigned to the watchful waiting arm of the SPCG-4 trial. The UK cohort consisted of 320 men with PSA-detected PCa in the Prostate testing for cancer and Treatment (ProtecT) study who opted for monitoring. Multilevel models describing changes in PSA level were fitted to the two cohorts, and average PSA level at age 50, change in PSA level with age, and predicted PSA values were derived. MEASUREMENTS: PSA level. RESULTS AND LIMITATIONS: In the SPCG-4 cohort, mean PSA at age 50 was similar to the cancer-free cohort but with a steeper yearly increase in PSA level (16.4% vs 4.0%). In the UK cohort, mean PSA level was higher than that in the cancer-free cohort (due to a PSA biopsy threshold of 3.0 ng/ml) but with a similar yearly increase in PSA level (4.1%). Predictions were less accurate for the SPCG-4 cohort (median difference between observed and predicted PSA level: -2.0 ng/ml; interquartile range [IQR]: -7.6-0.7 ng/ml) than for the UK cohort (median difference between observed and predicted PSA level: 0.8 ng/ml; IQR: -2.1-0.1 ng/ml). CONCLUSIONS: In PSA-detected men, yearly change in PSA was similar to that in cancer-free men, whereas in men with symptomatic PCa, the yearly change in PSA level was considerably higher. Our method needs further evaluation but has promise for refining active monitoring protocols. PMID- 19303696 TI - Editorial comment on: Development of a new method for monitoring prostate specific antigen changes in men with localised prostate cancer: a comparison of observational cohorts. PMID- 19303697 TI - High-dose chemotherapy plus stem cell transplantation in advanced germ cell cancer: a review. AB - CONTEXT: High-dose chemotherapy (HDCT) with stem cell transplantation (SCT) has been investigated as a treatment strategy for advanced germ cell cancer (GCC) for >2 decades. In an effort to improve on the overall cure rates of 80% achievable with conventional chemotherapy, researchers have investigated this therapeutic option as a first-line therapy for those with poor-prognosis disease and as a salvage therapy for those with relapsed or refractory disease. OBJECTIVE: The primary objective of this review is to define the role of HDCT plus SCT in advanced GCC. Prognostic indicators for this group of patients are also presented. EVIDENCE ACQUISITION: A Medline search of English-language literature was performed to identify studies published in the last 20 yr relating to the use of HDCT plus SCT in advanced GCC. Phase 1, phase 2, and phase 3 trials were included, as were retrospective reviews and meta-analyses. EVIDENCE SYNTHESIS: Phase 2 trials investigating HDCT plus SCT as a therapeutic option for advanced germ cell cancer have indicated a survival advantage over conventional chemotherapy. This has not been confirmed in the phase 3 setting. Alternative chemotherapeutic strategies and options following failure of HDCT plus SCT are discussed. CONCLUSIONS: Studies to date have not indicated a survival advantage for the use of HDCT plus SCT in advanced germ cell cancer. Many questions, however, remain unanswered, and further research is required to identify whether optimising the strategy of HDCT plus SCT will improve outcome in this predominantly young group of patients. PMID- 19303699 TI - Selective cytostatic and cytotoxic anticancer effects of bisfunctional agents: A strategy for the design of DNA binding agents. AB - Various agents have been synthesized and proved useful for the National Cancer Institute's anticancer testing as potential new drugs, but most agents suffer side effects from their limited selectivity against cancer cells over healthy ones. Therefore, this paper attempts to describe drugs in terms of the level of tumor cell selectivity which they possess to define the features of molecules that are essential for useful cytotoxicity. Selected cyclic amidinothymine analogues (NSC 697864, NSC 697865, and NSC 697869) have nanomolar inhibitory activities against leukemia cell lines: CCRF-CEM, HL-60(TB), while bisfunctional cancer fighters NSC 702408 and NSC 702409, showing larger numbers of cytostatic and cytotoxic effects, in an extended conformation would probably adopt a similar to NSC 715653 conformation leaving both opposite H-bond donor groups at the same distance to interact with DNA in a similar way. Such specific interactions (cell line selectivity to unique mutated patterns) lower considerably the observed dose response concentrations. This in vitro selectivity is shown to translate into in vivo efficacy indicated by the inflection in the cumulative testing curve. PMID- 19303700 TI - NF-kappaB inhibition in human hepatocellular carcinoma and its potential as adjunct to sorafenib based therapy. AB - Nuclear factor-kappaB (NF-kappaB) has been shown to play an important role in the development and progression of cancer. In this study, we systematically examined NF-kappaBp65 signaling pathway in both human hepatocellular carcinoma (HCC) tissue and HCC cell lines. NF-kappaBp65 signaling pathway is aberrantly expressed and activated in both human HCC tissue and HCC Hep3B cells. Inhibition of NF kappaB activity significantly reduced proliferation and invasion of Hep3B cells as well as down-regulated the expression of invasion-related molecules including matrix metalloproteinase (MMP)-2, MMP-9, membrane type-1 MMP (MT1-MMP), urokinase plasminogen activator (uPA) and vascular endothelial growth factor (VEGF). Hep3B cells exhibited a dose-dependent increase in apoptosis after receiving sorafenib treatment. Inhibition of NF-kappaB activity strongly sensitized Hep3B cells to sorafenib-induced cell death. Mechanistically, combined treatment of sorafenib and NF-kappaB inhibition enhanced inhibition of MAPK signaling and down regulation of anti-apoptotic protein Mcl-1 expression. These observations indicate that inhibition of NF-kappaB may be a potential antineoplastic therapy for HCC, especially the combination of NF-kappaB inhibition and sorafenib provides a novel therapeutic strategy for patients with advanced-stage HCC. PMID- 19303701 TI - Adsorption mechanism and property of a novel adsorption material PAM/SiO2 towards 2,4,6-trinitrotoluene. AB - 2,4,6-Trinitrotoluene (TNT) is toxic and mutagenic to many living organisms, so more and more rigorous limits on the letting amount of TNT have been established. In this paper, functional monomer acrylamide (AM) was grafted step by step on the surface of silica gel particles, and the grafted particle PAM/SiO(2) with strong adsorption ability for TNT was formed. The adsorption mechanism and properties of PAM/SiO(2) for TNT were researched through static and dynamic methods. The experimental results showed that PAM/SiO(2) possesses strong adsorption ability for TNT with interaction of three kinds of hydrogen bonds including peculiar NHcdots, three dots, centeredpi hydrogen bond (aromatic hydrogen bond) and CHcdots, three dots, centeredOC pi hydrogen bond. The saturated adsorption amount could reach to 0.873 mg g(-1). The empirical Freundlich isotherm was found to describe well the equilibrium adsorption data. In addition, the pH and temperature were found to have great influence on the adsorption amount. Mixture solution of HCl and ethanol is used as eluent, and the adsorbed TNT is eluted easily from PAM/SiO(2). Finally, PAM/SiO(2) was found to have excellent reusability. PMID- 19303702 TI - Recycling of non-metallic fractions from waste printed circuit boards: a review. AB - The major economic driving force for recycling of waste printed circuit boards (PCBs) is the value of the metallic fractions (MFs) of PCBs. The non-metallic fractions (NMFs), which take up almost 70wt% of waste PCBs, were treated by combustion or land filling in the past. However, combustion of the NMFs will cause the formation of highly toxic polybrominated dibenzodioxins and dibenzofurans (PBDD/Fs) while land filling of the NMFs will lead to secondary pollution caused by heavy metals and brominated flame retardants (BFRs) leaching to the groundwater. Therefore, recycling of the NMFs from waste PCBs is drawing more and more attention from the public and the governments. Currently, how to recycle the NMFs environmental soundly has become a significant topic in recycling of waste PCBs. In order to fulfill the better resource utilization of the NMFs, the compositions and characteristics of the NMFs, methods and outcomes of recycling the NMFs from waste PCBs and analysis and treatment for the hazardous substances contained in the NMFs were reviewed in this paper. Thermosetting resin matrix composites, thermoplastic matrix composites, concrete and viscoelastic materials are main applications for physical recycling of the NMFs. Chemical recycling methods consisting of pyrolysis, gasification, supercritical fluids depolymerization and hydrogenolytic degradation can be used to convert the NMFs to chemical feedstocks and fuels. The toxicity characteristic leaching procedure (TCLP) and synthetic precipitation leaching procedure (SPLP) can be used to determine the toxicity characteristic (TC) of the NMFs and to evaluate the environmental safety of products made from the recycled NMFs. It is believed that physical recycling of the NMFs has been a promising recycling method. Much more work should be done to develop comprehensive and industrialized usage of the NMFs recycled by physical methods. Chemical recycling methods have the advantages in eliminating hazardous substances in the NMFs. The trend in chemical recycling of the NMFs is to make the best of advantages over physical recycling of the NMFs to compensate its higher cost. Removing and treating the hazardous substances in the NMFs is an ultimate method to eliminate the pollution. PMID- 19303703 TI - Characterization of organic matter and disinfection by-products in membrane backwash water from drinking water treatment. AB - Two pilot-scale membrane plants were set up to produce drinking water, and membrane backwash water was discharged during the production process. This work studied the characteristics of dissolved organic matter (DOM) in membrane backwash water from submerged microfiltration (MBWS) and pressurized ultrafiltration (MBWP) both of which are coupled with the pre-coagulation process. The results showed that the two waters had similar molecular weight (MW) distributions. Dissolved organic carbon (DOC) and trihalomethane formation potential (THMFP) in MBWS and MBWP were both mainly distributed in MW>30 kDa and MW<1 kDa, and UV(254) was mainly in MW<1 kDa. For Luan River water (LRW, the raw water for the two pilot-scale membrane plants in this study), organic matter enriched in membrane backwash water was mainly in sizes of MW>30 kDa. In addition, organic matter with MW>10 kDa was higher in MBWP than in MBWS. The quality of membrane backwash water was influenced by the changes in LRW quality during different periods. The quality of membrane backwash water was worse in alga-laden period than in normal period and organic matter concentrations in MW<1 kDa increased significantly in this period. The small size DOM in membrane backwash water was more reactive to form trihalomethanes (THMs) in the disinfection process. The variability of specific UV absorbance and THMFP/DOC was consistent in membrane backwash water. PMID- 19303704 TI - Material and structural characterization of alkali activated low-calcium brown coal fly ash. AB - The waste low-calcium Czech brown coal fly ash represents a considerable environmental burden due to the quantities produced and the potentially high content of leachable heavy metals. The heterogeneous microstucture of the geopolymer M(n) [-(Si-O)(z)-Al-O](n).wH(2)O, that forms during the alkaline activation, was examined by means of microcalorimetry, XRD, TGA, DSC, MIP, FTIR, NMR MAS ((29)Si, (27)Al, (23)Na), ESEM, EDS, and EBSD. The leaching of heavy metals and the evolution of compressive strength were also monitored. The analysis of raw fly ash identified a number of different morphologies, unequal distribution of elements, Fe-rich rim, high internal porosity, and minor crystalline phases of mullite and quartz. Microcalorimetry revealed exothermic reactions with dependence on the activator alkalinity. The activation energy of the geopolymerization process was determined as 86.2kJ/mol. The X-ray diffraction analysis revealed no additional crystalline phases associated with geopolymer formation. Over several weeks, the (29)Si NMR spectrum testified a high degree of polymerization and Al penetration into the SiO(4) tetrahedra. The (23)Na NMR MAS spectrum hypothesized that sodium is bound in the form of Na(H(2)O)(n) rather than Na(+), thus causing efflorescence in a moisture-gradient environment. As and Cr(6+) are weakly bonded in the geopolymer matrix, while excellent immobilization of Zn(2+), Cu(2+), Cd(2+), and Cr(3+) are reported. PMID- 19303705 TI - Biosorption of uranium(VI) from aqueous solution using calcium alginate beads. AB - In this paper, sorption potentials of uranium ions were studied using alginate polymer beads in diluted aqueous solutions. The ability of alginate beads to adsorb uranium(VI) from aqueous solution has been studied at different optimized conditions of pH, U(VI) concentration, contact time, biomass dosage and temperature. In order to determine the adsorption characteristics, Langmuir, Freundlich, and Dubinin-Radushkevich adsorption isotherms were applied to the adsorption data. The thermodynamic parameters such as variations of enthalpy DeltaH, entropy DeltaS and variation of Gibbs free energy DeltaG were calculated from the slope and intercept of lnK(d) vs. 1/T plots. The results suggested that alginate beads could be suitable as a sorbent material for adsorption and removal of uranium ions from dilute aqueous solutions. PMID- 19303706 TI - Formation of PCDD and PCDF in the thermal treatment of footwear leather wastes. AB - The leather waste generated by the footwear industry is considered dangerous due to the presence of trivalent chromium, derived from the salt utilized to tan hides. In Brazil, the majority of this waste is disposed on landfills and only about 3% are recycled. The thermal treatment is an alternative method for purification of such residues. By using this technique it is possible to generate energy and recover the chromium present in the ash for the production of basic chromium sulfate (tanning industry), high carbon ferrochromium or carbon-free ferrochromium (steel industry). In the last 10 years, the gasification and combustion of footwear leather waste have been intensively studied at the Federal University of Rio Grande do Sul. The research experiment for characterization of the emissions of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) were carried out in a semi-pilot unit (350 kW(th)). From new investments the thermal capacity of the unit will increase to 600 kW(th). The unit will produce power from the heat generated in the combustion. The experimental results indicated that during the thermal treatment of footwear leather wastes, the formation mechanism of PCDD/F is the de novo synthesis. Most of PCDD/F were found in the particulate phase (>95%). A kinetic model was used for discussion of the achieved experimental results. The model is based in the carbon gasification, PCDD/F formation, desorption and degradation. From the conclusions obtained in this work will be possible minimize the PCDD/F formation in process of combustion of footwear leather wastes. PMID- 19303707 TI - Selectivity of cesium from fission radionuclides using resorcinol-formaldehyde and zirconyl-molybdopyrophosphate as ion-exchangers. AB - Two ion-exchangers, the first is organic in nature and the other is inorganic named as resorcinol-formaldehyde (R-F) and zirconyl-molybdopyrophosphate (ZMPP) were synthesized and applied to decontaminate some fission radionuclides from radioactive waste streams. The distribution coefficients of the studied radionuclides for the two ion-exchanges at varying pH conditions are evaluated and the data showed that the selectivity sequence of resorcinol-formaldehyde for the studied cations is Cs(+)>Co(2+)>Eu(3+)>Zn(2+) which is the same sequence of zirconyl-molybdopyrophosphate while as the X-ray diffraction characterization showed that resorcinol-formaldehyde resin is an amorphous material but zirconyl molybdopyrophosphate is a semicrystalline one and both of resorcinol-formaldehyde and zirconyl-molybdopyrophosphate are considered as promising materials specially for the decontamination of cesium from radioactive waste solutions. PMID- 19303708 TI - The effect of EDDS addition on the phytoextraction efficiency from Pb contaminated soil by Sedum alfredii Hance. AB - Present study reports the results of three pot experiments, conducted to investigate the chelate-assisted phytoextraction of Pb contaminated soils. The optimum phytoextraction was observed when 2.5mM ethylene diamine disuccinic acid (EDDS) was added in single dosage for 14 days to low Pb soil (treated with 400 mg kg(-1)soil). On the contrary, for high Pb soil (treated with 1200 mg kg(-1)soil), 5mM EDDS concentration in single dosage for 10 days produced better results. Post harvest effects of EDDS on the concentrations of available Pb and dissolved organic carbon (DOC) were significantly higher as compared with check (CK i.e. without EDDS addition), and consequently decreased with the passage of time. Our results suggested that chelate-assisted phytoextraction was more suitable for slightly contaminated soils. PMID- 19303709 TI - Treatment of Terasil Red R dye wastewater using H2O2/pyridine/Cu(II) system. AB - The H(2)O(2)/pyridine/Cu(II) advanced oxidation system was used to assess the efficiency of the treatment of a 1 g L(-1) Terasil Red R dye solution. This system was found to be capable in reducing the concentration of chemical oxygen demand (COD) of the dye solution up to 90%, and achieving 99% in decolorization at the optimal concentration of 5.5mM H(2)O(2), 38 mM pyridine and 1.68 mM Cu(II). The final concentration of COD was recorded at 117 mg L(-1) and color point at 320 PtCo. Full 2(4) factorial design and the response surface methodology using central composite design (CCD) were utilized in the screening and optimization of this study. Treatment efficiency was found to be pH independent. The amount of sludge generation was in the range of 100-175 mg L(-1) and the sludge produced at the optimal concentration was 170 mg L(-1). PMID- 19303710 TI - Near-grain-boundary characterization by atomic force microscopy. AB - Characterization of near-grain boundary is carried out by atomic force microscopy (AFM). It has been observed to be the most suitable technique owing to its capability to investigate the surface at high resolution. Commercial purity-grade nickel processed under different conditions, viz., (i) cold-rolled and annealed and (ii) thermally etched condition without cold rolling, is considered in the present study. AFM crystallographic data match well with the standard data. Hence, it establishes two grain-boundary relations viz., plane matching and coincidence site lattice (CSL Sigma=9) relation for the two different sample conditions. PMID- 19303711 TI - Detection of a Hepatozoon and spotted fever group Rickettsia species in the common marsupial tick (Ixodes tasmani) collected from wild Tasmanian devils (Sarcophilus harrisii), Tasmania. AB - Tasmanian devils are the largest extant marsupial carnivores, confined to the Australian island state of Tasmania. The iconic marsupial has dramatically declined in number since the discovery of devil facial tumor disease in 1996 and efforts are being made to uncover vital information to assist in the long-term survival of the species. Ticks are the main vectors of arthropod-borne disease in animals, raising the question of whether Tasmanian devils may be host to arthropods capable of harboring infectious agents. Partially engorged ticks were collected from 35 wild Tasmanian devils and tested for the presence of a range of tick-borne genera. A spotted fever group Rickettsia was detected in 45.5% of samples of the tick Ixodes tasmani (n=44), from all trapping locations, sharing close sequence identity to members of the Rickettsia massiliae group. A Hepatozoon species was also identified in 34.1% of the same sample set, sharing sequence similarities to Hepatozoon felis, a known pathogen of felids. Dual detection was identified in 13.6% of tick samples, where prevalence of the two genera overlapped. The existence of two previously undetected species of genera known for containing pathogens identifies additional potential risks to the health of the devil population. PMID- 19303712 TI - Sprayed cultured autologous keratinocytes used alone or in combination with meshed autografts to accelerate wound closure in difficult-to-heal burns patients. PMID- 19303713 TI - Preventable irreversible side effects of critical socioeconomic decisions on burn. PMID- 19303714 TI - Evaluation of the spontaneous breathing trial in burn intensive care patients. AB - BACKGROUND: The extubation failure rate in our burn patients is 30%. OBJECTIVE: To evaluate the influence of the 30 min spontaneous breathing trial on extubation outcome in burn patients. METHODS: A prospective, observational study in a burn intensive care unit. All adult patients requiring mechanical ventilation for >24h and meeting the inclusion criteria underwent a 30 min spontaneous breathing trial (SBT). Extubation was undertaken after a successful SBT. RESULTS: Of 49 planned extubations, 9 failed (18%), much lower than the 30% extubation failure rate identified prior to the implementation of the SBT. The duration of ventilation was significantly shorter (p=0.04) in the patients who passed a SBT and those who failed extubation were significantly older (p=0.003). The logistic regression analysis identified that age independently predicted extubation outcome. Patients who failed extubation, after a successful SBT, had a significantly longer duration of ventilation (p=0.0001) and ITU length of stay (p=0.001). CONCLUSIONS: The incidence of extubation failure was much lower and the duration of ventilation significantly shorter in patients who were extubated after a successful SBT. These findings support the use of the SBT in burn patients. Age independently predicts extubation outcome in burn patients who have passed a SBT. PMID- 19303715 TI - A new diagnostic tool for the classification of patients with self-inflicted burns (SIB): the SIB-Typology and its implications for clinical practice. AB - Although self-inflicted burns comprise a rather small percentage of patients treated in burn units, they represent a challenging subgroup for the specialist team. According to the literature, these patients, often with a past psychiatric history, remain in the hospital longer due to delayed wound healing, increased number of operative sessions and reduced compliance. The aim of this study was to analyse the incidence of psychiatric disorders and the overall outcome in patients with self-inflicted burns admitted to the Vienna Burn Centre in the past 11 years. We therefore reviewed the following parameters: past psychiatric history, burn extent, burn depth, mechanism of burn, treatment, length of hospital stay, and mortality. Based on our evaluation patients can be grouped into three distinct categories based on a simple two-axis diagram, the Self Inflicted-Burns-Typology (SIB-T), including the parameters "psychiatric disorder" and "suicide attempt": "typical", "delirious" and "reactive". These three groups seem to differ in treatment outcome and later course of rehabilitation. PMID- 19303716 TI - Effect of ablated bronchial blood flow on survival rate and pulmonary function after burn and smoke inhalation in sheep. AB - The bronchial circulation plays a significant role in the pathophysiological changes of burn and smoke-inhalation injury. Bronchial blood flow markedly increases immediately after inhalational injury. This study examines whether the ablation of the bronchial artery attenuates pathophysiological changes and improves survival after burn and smoke-inhalational injury in an ovine model. Acute lung injury was induced by 40% total body surface-area third-degree cutaneous burn and cotton smoke inhalation (48 breaths of cotton smoke, <40 degrees C) under deep anaesthesia. Twelve adult female sheep were divided into two groups: (1) sham (injured, non-ablated bronchial artery, n=6); (2) ablation (injured, ablated bronchial artery, n=6). Ablation of the bronchial artery was performed 72 h before the injury. The experiment was continued for 96 h. Burn and smoke-inhalation injury significantly increased regional blood flow in the bronchi. Ablation of the bronchial artery significantly reduced acute regional blood flow increases in the proximal and distal bronchi. All animals in the ablation group survived to 96 h. Four of these were successfully weaned off the ventilator. Three animals of the sham group met standardised euthanasia criteria at 60 h, while another met the criteria at 78 h. The lung wet-to-dry weight ratio, histology score and myeloperoxidase (MPO) activity were significantly increased by the insult, but ablation of the bronchial artery attenuated these changes. Burn and smoke-inhalation injury induced a significant increase in bronchial blood flow and accelerated airway obstruction, pulmonary vascular changes, pulmonary oedema and pulmonary dysfunction. Ablated bronchial circulation attenuated these pathophysiological changes. PMID- 19303717 TI - Extramammary Paget's disease arising from a burn scar of right forearm: An uncommon case report. PMID- 19303718 TI - Accuracy of references in burns journals. AB - AIMS: To study the incidence and risk factors for citation and quotation errors in two major burns surgery journals. METHODS: 120 references were randomly selected from original articles published in the following two journals - January to December 2006 issues of Burns and Journal of Burn Care & Research. For each reference, the ease of retrieval on PubMed and the presence of citation errors were noted. Two independent observers analysed each reference for quotation errors. The characteristics of the root article, that is, type of study, author numbers, number of references and article word count were noted. RESULTS: Of the 120 selected references, 117 referred to articles from indexed medical journals published in English. Among these, 4 articles could not be retrieved due to fatal citation errors (3.3%). A further 12 citation errors were noted giving a total citation error rate of 13.3% (95% CI: 6.74-19.93%). Of the 117 references analysed, the quotation error rate was 13.7% (95% CI: 8.6-19.5%) half of which were major errors. There was no significant association between the combined error rate per article and the journal (Kruskal-Wallis test; p=0.861, type of study (Kruskal-Wallis test; p=0.717), author numbers (Spearman's rho=0.197, p=0.423), article length (Spearman's rho=0.118, p=0.705) or references per article (Spearman's rho=0.229, p=0.189). CONCLUSION: Significant numbers of citation and quotation errors still appear in current burns literature. Incorrect spelling of author names and partial omissions of article titles were the two most common errors. No observable underlying factors were identified in this study. The present results serve as a reminder to authors, editors and peer reviewers for more care of citation accuracy when striving for their common goal of scientific excellence. PMID- 19303719 TI - An interesting reason for expander failure: leakage from the base of the port. PMID- 19303721 TI - Erectile function durability following permanent prostate brachytherapy. AB - PURPOSE: To evaluate long-term changes in erectile function following prostate brachytherapy. METHODS AND MATERIALS: This study included 226 patients with prostate cancer and preimplant erectile function assessed by the International Index of Erectile Function-6 (IIEF-6) who underwent brachytherapy in two prospective randomized trials between February 2001 and January 2003. Median follow-up was 6.4 years. Pre- and postbrachytherapy potency was defined as IIEF-6 > or = 13 without pharmacologic or mechanical support. The relationship among clinical, treatment, and dosimetric parameters and erectile function was examined. RESULTS: The 7-year actuarial rate of potency preservation was 55.6% with median postimplant IIEF of 22 in potent patients. Potent patients were statistically younger (p = 0.014), had a higher preimplant IIEF (p < 0.001), were less likely to be diabetic (p = 0.002), and were more likely to report nocturnal erections (p = 0.008). Potency preservation in men with baseline IIEF scores of 29-30, 24-28, 18-23, and 13-17 were 75.5% vs. 73.6%, 51.7% vs. 44.8%, 48.0% vs. 40.0%, and 23.5% vs. 23.5% in 2004 vs. 2008. In multivariate Cox regression analysis, preimplant IIEF, hypertension, diabetes, prostate size, and brachytherapy dose to proximal penis strongly predicted for potency preservation. Impact of proximal penile dose was most pronounced for men with IIEF of 18-23 and aged 60-69. A significant minority of men who developed postimplant impotence ultimately regained erectile function. CONCLUSION: Potency preservation and median IIEF scores following brachytherapy are durable. Thoughtful dose sparing of proximal penile structures and early penile rehabilitation may further improve these results. PMID- 19303720 TI - Neonatal overfeeding alters adult anxiety and stress responsiveness. AB - The neonatal nutritional environment is critical for programming the systems regulating body weight, and an inappropriate nutritional environment in early life can lead to overweight or obesity later on. In this study we demonstrate that changes to the neonatal nutritional environment, induced by changing the litter size in which the animal is raised, can alter not only body weight into adulthood, but also anxiety behaviours and stress responsiveness. These effects appear to be somewhat sex-dependent, affecting female rats more strongly than males. Thus, Wistar rats raised in small litters, where they have greater access to their mothers' milk, weigh more in adulthood than those raised in large litters. Females from these small litters show enhanced exploratory behaviour and reduced anxiety in the elevated plus maze, entering the open arms more often. They also display enhanced central responses to restraint stress including greater activation of the paraventricular nucleus of the hypothalamus and paraventricular nucleus of the thalamus, possibly indicating that the changes we see are related to enhanced arousal. Thus, while neonatal litter size affects long-term body weight regulation, it can also alter long-term activity, anxiety and stress responsiveness, and the degree to which it does so appears to be sex dependent. PMID- 19303722 TI - Association of NAT2, GSTM1, GSTT1, CYP2A6, and CYP2A13 gene polymorphisms with susceptibility and clinicopathologic characteristics of bladder cancer in Central China. AB - OBJECTIVE: To explore the association of polymorphisms in N-acetyltransferase 2 (NAT2), glutathione S-transferase (GST), cytochrome P450 (CYP) 2A6, and CYP 2A13 genes with susceptibility and clinicopathologic characteristics of bladder cancer in a Chinese population. METHODS: In a hospital-based case-control study of 208 cases and 212 controls matched on age and gender, genotypes were determined by PCR-based methods. Risks were evaluated by unconditional logistic regression analysis. RESULTS: It was found that significant associations of the NAT2 slow acetylator genotype (odds ratio, OR: 2.42; 95% confidence interval, CI: 1.47 3.99), GSTM1 null genotype (OR: 1.64; 95% CI: 1.11-2.42) and GSTM1/GSTT1-double null genotype (OR: 1.72; 95% CI: 1.00-2.95) with increased risk of bladder cancer. Conversely, carriers with at least one CYP2A6*4 allele showed lower risk than the non-carriers (OR: 0.47; 95% CI: 0.28-0.79). The adjusted ORs (95% CI) for smokers with NAT2 slow-acetylator, GSTM1 null, GSTM1/GSTT1-double null genotype, and variant CYP2A6 genotypes were 2.99 (1.44-6.25), 1.98 (1.13-3.48), 2.66 (1.22-5.81) and 0.41 (0.20-0.86), respectively. Furthermore, NAT2 slow acetylator, GSTM1 null, and GSTM1/GSTT1-double null genotypes were associated with higher tumor grade (P=0.001, 0.022, and 0.036, respectively), and only NAT2 slow-acetylator genotype was associated with higher tumor stage (P=0.007). CYP2A13 was not associated with risk or tumor characteristics. CONCLUSION: It is suggested that NAT2 slow-acetylator, GSTM1 null, GSTM1/GSTT1-double null, and variant CYP2A6 genotypes may play important roles in the development of bladder cancer in Henan area, China. PMID- 19303723 TI - Cardiovascular and subjective effects of repeated smoked cocaine administration in experienced cocaine users. AB - Studies using rodents have shown that behavioral responses to a stimulant are enhanced when the stimulant is given within the same context as previous stimulant administrations; this increase in effect related to context is often referred to as sensitization. We examined the role of environmental stimuli in modulating the subjective and cardiovascular effects of cocaine in humans (1) within a daily "binge" and (2) after cocaine abstinence. Ten non-treatment seeking users of smoked cocaine were admitted to the hospital for 17 consecutive days. Participants smoked cocaine (25mg/dose) under two counterbalanced conditions: paired stimuli (same stimuli presented each session) and unpaired stimuli (varied stimuli presented each session). Under each stimulus condition, participants had cocaine test sessions for three consecutive days, no sessions for the next 3 days, then another cocaine test session on the following day, for a total of eight test days. Stimulus condition had no effect on cardiovascular or subjective effects so data were analyzed as a function of repeated cocaine administration over 2 weeks. Maximal ratings on "good drug" and "drug rating" subjective effects clusters decreased over days of repeated cocaine exposure. In contrast, baseline and peak heart rate and systolic pressure increased over days of repeated cocaine administration. Thus, repeated administration of smoked cocaine to experienced cocaine users resulted in increases in baseline blood pressure and heart rate and modest decreases in positive subjective effects. These data indicate modest tolerance rather than sensitization to the positive subjective effects of cocaine with repeated exposure. PMID- 19303724 TI - Multiple typing for the epidemiological study of contamination of broilers with thermotolerant Campylobacter. AB - This study aims to investigate the genetic diversity of thermotolerant Campylobacter in commercial broiler flocks and in the environment of broiler farms in Belgium. Seven out of 18 investigated flocks became colonized during rearing. Fluorescent amplified fragment length polymorphism (FAFLP), pulsed field gel electrophoresis (PFGE), restriction fragment length polymorphism of the flagellin A gene (flaA-RFLP) and antimicrobial resistance profile (ARP) were used for typing of the isolates. By the combination of FAFLP and PFGE, 22 Campylobacter genotypes could be distinguished. Colonization was almost exclusively with Campylobacter jejuni and unique genotypes were found in each flock. Multiple genotypes were detected in the broilers of 3 flocks, either simultaneously or successively. In 5 flocks, strains that were resistant to at least one antibiotic (mostly tetracycline) were found. The presence of other broiler houses on the farm did not result in a higher probability of colonization. The nipple water was contaminated with the same genotype as the broilers, illustrating its importance for transmission of Campylobacter. The same genotype was detected in a water puddle and in the broiler flock during rearing in 3 flocks. Once, the same genotype was isolated from the ditch water shortly before it was detected in the broilers. PMID- 19303725 TI - Genetic organization and preferential distribution of putative pilus gene clusters in Streptococcus suis. AB - Recent analyses of Streptococcus suis isolates using multilocus sequence typing (MLST) suggested the importance of sequence type (ST) 1 and ST27 complexes for animal hygiene and public health. In this study, to investigate whether pilus associated genes in S. suis can be used as novel genetic markers for important clonal groups, we examined the correlation between STs and putative pilus associated gene profiles in S. suis. Genomic searches using sequenced genomes and sequence data determined in several isolates revealed the presence of at least four distinct putative pilus gene clusters in S. suis (srtBCD, srtE, srtF, and srtG clusters). On the basis of the presence or absence of genes in the four clusters, 108 S. suis isolates from various origins were classified into 12 genotypes (genotypes A-L). Genotypes A and B, which possessed srtBCD plus srtF clusters and srtF plus srtG clusters, respectively, were the most common in isolates from diseased pigs and humans, and 29.9% and 59.8% of the isolates belonged to genotypes A and B, respectively. In contrast, only 4.8% and 28.6% of isolates from healthy carriers were genotypes A and B, respectively. MLST analysis showed the associations of genotypes A and B with ST1 and ST27 complexes, respectively. In addition, srtBCD and srtG clusters were preferentially distributed to ST1 and ST27 complex members, respectively. These results suggest that profiling of selected pilus-associated genes could be used as an easy screening method to monitor isolates important for S. suis infection. PMID- 19303726 TI - Application of environmental magnetism on crime detection in a highway traffic accident from Yangzhou to Guazhou, Jiangsu Province, China. AB - Both elemental composition analysis and mineral magnetic measurements were used to infer the source of the earth dumped on a highway in China, which resulted in human life loss and injury in 2002. The results indicate that the earth and the samples from the potential two sources are very similar in geochemical compositions. However, magnetic properties show stronger difference among the samples. A plot of magnetic susceptibility (chi) vs. Anhysteretic Remanent Magnetization (ARM) clearly show that the earth at the accident site (sample No. 1) is closely matched with one sample (No. 4) from the source site B. Such a difference in geochemical and magnetic signatures among the samples is ascribed to the nature of the earth, which is derived from the Xiashu Loess in southern China. Our results indicate that environmental magnetic method can help to crime investigation by aiding in the provenance tracing of soils/sediments evidences in a simple, economic and non-destructive way. PMID- 19303727 TI - An autopsy case presenting repetitive hypoglycemia and unique cortical dysplasia. AB - In epileptic patients, focal cortical dysplasia (FCD) is pathologically characterized by irregular cortical lamination, blurring of the grey and white matter border and the occurrence of dysplastic cells in the cerebral cortex. Here, we report the case of a 42-year-old male showing developmental delay, transient repetition of hypoglycemic attack and cortical dysplasia, partly mimicking FCD. He had no family history of neurological disorders. He had never been able to stand independently and had always been unable to speak. He developed generalized convulsion in infancy, and then, in the absence of predisposing factors, suffered from repetitive hypoglycemic attacks between the ages of 27 and 38. Various endocrine tests, abdominal CT and brain MRI failed to demonstrate abnormalities. He died of peritonitis. At autopsy, no changes were observed in the pancreas, liver, kidneys, endocrine organs or hypothalamus. In the insular and frontal cortices, many large bizarre cells in the deep layer were observed and perivascular oligodendrocyte satellitosis was present in the adjacent white matter. Unlike FCD, the cortical lamination and the grey-white matter interface were preserved. A well-demarcated pilocytic astrocytoma was present in the brainstem. The cortical dysplasia, consisting of the diffuse occurrence of bizarre cells and the preservation of cortical lamination, is unique and has not been previously reported. Repetition of hypoglycemic attacks within a certain period is also noteworthy, although the relationship of this with the cortical dysplasia is unknown. PMID- 19303728 TI - Factors influencing influenza-vaccination in adults under immunosuppressive therapy for a systemic inflammatory disease. AB - OBJECTIVE: The aim of this study was to evaluate flu vaccination rates and influencing factors in patients with systemic inflammatory diseases. METHODS: All patients presenting with a systemic inflammatory disease and taking immunosuppressants, who were hospitalized or had consulted in our internal medicine department between January 2 and 31, 2006, were included in the study. The information concerning flu vaccination was collected with a standardized form. RESULTS: One hundred and thirty-seven patients (mean age 53.1+/-17.6years; 40 [29%] male patients) were included: 39 (28%) had received flu vaccination in 2005 including 14 (16.7%) of the 84 patients with no other indication for flu vaccination than IS-induced immunodepression and 25 (47.2%) of the 53 patients with other flu vaccination indication(s) (p<0.001). The most frequent reasons for non-vaccination were: absence of physician recommendation (58%), fear of adverse effects (35%) and concern on vaccine clinical effectiveness (5%). The vaccination rate was significantly higher (49%) among patients who remembered having received a voucher from the French National Health Insurance Agency versus 18% among those who did not (OR=4.2 [95%CI, 1.92-9.19] p<0.05). This correlation remained significant after adjustment for confounding factors in a logistic regression model. CONCLUSION: Influenza-vaccination coverage is low in patients receiving immunosuppressive therapy for systemic inflammatory diseases. We have to increase the influenza-vaccination coverage in this population. PMID- 19303729 TI - [Therapeutic EUS for the management of pancreatic and biliary diseases]. PMID- 19303730 TI - From protein-protein interaction to therapy response: molecular imaging of heat shock proteins. AB - HSP70 promoter-driven gene therapy and inhibition of HSP90 activity with small molecule inhibitors are two shining points in a newly developed cohort of cancer treatment. For HSP70 promoters, high efficiency and heat inducibility within a localized region make it very attractive to clinical translation. The HSP90 inhibitors exhibit a broad spectrum of anticancer activities due to the downstream effects of HSP90 inhibition, which interfere with a wide range of signaling processes that are crucial for the malignant properties of cancer cells. In this review article, we summarize exciting applications of newly emerged molecular imaging techniques as they relate to HSP, including protein protein interactions of HSP90 complexes, therapeutic response of tumors to HSP90 inhibitors, and HSP70 promoters-controlled gene therapy. In the HSPs context, molecular imaging is expected to play a vital role in promoting drug development and advancing individualized medicine. PMID- 19303731 TI - Molecular diversity of Bacteroides spp. in human fecal microbiota as determined by group-specific 16S rRNA gene clone library analysis. AB - Bacteroides spp. represent a prominent bacterial group in human intestinal microbiota with roles in symbiosis and pathogenicity; however, the detailed composition of this group in human feces has yet to be comprehensively characterized. In this study, the molecular diversity of Bacteroides spp. in human fecal microbiota was analyzed from a seven-member, four-generation Chinese family using Bacteroides spp. group-specific 16S rRNA gene clone library analysis. A total of 549 partial 16S rRNA sequences amplified by Bacteroides spp. specific primers were classified into 52 operational taxonomic units (OTUs) with a 99% sequence identity cut-off. Twenty-three OTUs, representing 83% of all clones, were related to 11 validly described Bacteroides species, dominated by Bacteroides coprocola, B. uniformis, and B. vulgatus. Most of the OTUs did not correspond to known species and represented hitherto uncharacterized bacteria. Relative to 16S rRNA gene universal libraries, the diversity of Bacteroides spp. detected by the group-specific libraries was much higher than previously described. Remarkable inter-individual differences were also observed in the composition of Bacteroides spp. in this family cohort. The comprehensive observation of molecular diversity of Bacteroides spp. provides new insights into potential contributions of various species in this group to human health and disease. PMID- 19303732 TI - Rapid resolution liquid chromatography-mass spectrometry determination of SAR97276 in monkey matrices. Pharmacokinetics in rhesus monkey infected by Plasmodium cynomolgi. AB - Since several years, we developed a new class of antimalarial drugs targeting the phospholipid metabolism of the Plasmodium falciparum malaria parasite. The bis thiazolium compound, SAR97276, is the lead compound and is now in clinical development. In this paper, we applied the fast rapid resolution liquid chromatography-mass spectrometry technique to the analysis of SAR97276 in monkey matrices. The sample pre-treatment procedure involved an acidic precipitation of proteins followed by solid-phase extraction. The monocationic compound, T2, was used as internal standard. A good separation was achieved on a Zorbax eclipse XDB C8 column (1.8 microm, 50 mm x 4.6mm) with a mobile phase consisting of acetonitrile-trimethylamine-formate buffer (pH 3) gradient elution. The total run time was 8 min. Inter-assay precisions were <10% in plasma, and 85% in plasma, and >75% in blood. The lower limits of quantitation were 3.3 microg/l in plasma and 3.3 microg/kg in blood. No matrix effect was observed. This newly developed method is sensitive, selective, reproducible, and stability indicating. It was used to analyse samples taken during a pharmacokinetic/pharmacodynamic study carried out in infected Rhesus monkey by Plasmodium cynomolgi as part of the ongoing development of SAR97276. PMID- 19303733 TI - Alcohol management strategies of college students with diabetes. AB - OBJECTIVE: Alcohol management strategies of college undergraduates with diabetes were studied in order to identify which strategies were associated with decreased alcohol consumption and consequences. METHODS: Data came from 450 undergraduates with diabetes who were among 90,000 students from 123 postsecondary institutions who completed the Spring, 2006 National College Health Assessment. RESULTS: A majority of respondents (68.0%) reported alcohol use in the previous month, with 41.8% consuming five or more drinks in one sitting during the previous 2 weeks. High alcohol management strategy use in the past year was associated with fewer heavy drinking episodes and fewer alcohol-related consequences. Two strategies, avoiding drinking games and pacing one's drinking, were especially strong predictors of reduced consumption and consequences, and were more common among older students. CONCLUSION: Developing effective strategies to manage alcohol intake is especially important for students with diabetes due to short- and long term hazards of excessive drinking. PRACTICAL IMPLICATIONS: Alcohol use is a common aspect of college culture and may hold perceived social benefits for students. Healthcare clinicians should help students with diabetes to identify alcohol management strategies that are perceived as effective and feasible. Avoiding drinking games and pacing drinks appear to be strategies especially worthy of promotion. PMID- 19303735 TI - WITHDRAWN: Effect of cilostazol in alleviating cardiovascular complications through regulation of type 1 plasminogen activator inhibitor and transforming growth factor-beta1 overexpression in experimental rats. AB - This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. PMID- 19303734 TI - [Venous thromboprophylaxis postcaesarean: survey of practices in Lorraine]. PMID- 19303736 TI - [Renal aging: risk factors and nephroprotection]. AB - Renal failure in the elderly is currently underestimated and presents a real challenge for the public health system. Kidney function must be routinely assessed by creatinine clearance, estimated with either the Cockcroft and Gault formula or the simplified MDRD formula, which appears especially appropriate for the elderly. Normal kidney aging is related to tissue and functional changes that make older patients very vulnerable to environmental modifications. Numerous factors can accelerate the impairment of rental function during aging. Some of them cannot be modified: sex, genome, and initial kidney disease. Most of them can be managed or treated: hypertension, diabetes mellitus, obesity, smoking, dyslipidemia, proteinuria, and the presence of oxidation and glycation products. Chronic renal failure in the elderly must be managed early with strict treatment targets to avoid the development of end-stage renal disease. Inhibitors of the renin-angiotensin-aldosterone system play an essential role in optimizing nephroprotection: control of hypertension, diabetes complications, and proteinuria. They should be prescribed very carefully in older patients. Age is not a prerequisite for consultations with nephrologists, which should take place early so that nephroprotection can still be useful. PMID- 19303737 TI - [Biotherapy of auto-immune diseases : past, present and future perspectives]. AB - During the last decade, new biotherapies have been developed for the treatment of systemic autoimmune diseases. These new approaches are based on a better understanding of the auto-immune response. Targets of these new treatments are the main steps of the immune response. These new treatments include: B lymphocytes inhibitors, inhibitors of the costimulation between antigen presenting cells and T lymphocytes, T lymphocytes inhibitors, B and T lymphocytes inhibitors, modulators of the cytokine network, complement system inhibitors, and chemokines and integrins inhibitors. Advances in molecular biology, genetics and immunology have accelerated our understanding of auto-immune diseases, and allow identification of new targets. Development of new biotechnological tools with longer half-lives and increased affinity is an important next step. In spite of these exciting new perspectives, a systematic risk/benefit evaluation of these new treatments appears mandatory because immunity plays a key role in the control of infectious and tumor diseases. Further research is also needed to evaluate the cost/effectiveness ratio of these new therapies. PMID- 19303738 TI - Iliopsoas tendonitis due to the protrusion of an acetabular component fixation screw after total hip arthroplasty. AB - Postoperative pain after total hip arthroplasty can have a wide range of underlying causes. Iliopsoas tendonitis secondary to the impingement of this tendon is a relatively rare cause of pain after arthroplasty. This condition is characterized by pain on active flexion and an absence of signs or symptoms of loosening or infection. In this report, we describe the case of a patient who had signs and symptoms of iliopsoas tendonitis secondary to the protrusion of an acetabular fixation screw through the ilium after primary total hip arthroplasty. Nonoperative treatment was ineffective, and the patient ultimately underwent surgical removal of the screw. The severity of the patient's symptoms decreased significantly after the operation. PMID- 19303739 TI - Normative voice range profiles in vocally trained and untrained children aged between 7 and 10 years. AB - Only limited data on normal vocal constitution and vocal capabilities in school aged children are available. To take better care of children's voices, it might be helpful to know voice ranges and limits of not only vocally trained but also vocally untrained children. Goal of this study was the evaluation of singing voice capabilities of vocally healthy children with different social and vocal/musical backgrounds using voice range profile measurements (VRP). VRP percentiles that reflect constitutional aspects were suggested. In this cross sectional study, 186 children (aged between seven and 10 years), attending five schools, were included. VRP measurements were performed under field conditions. Interviews and questionnaires regarding vocal strain and vocal training were applied; the answers were used for classification of singing activity and vocal training (KLASAK). All children reached a mean singing voice range of at least two octaves. By using the answers of interviews and questionnaires, the children could be classified according to vocal strain and vocal training. The groups showed no significant differences regarding VRP measurements. In the following step, percentiles were calculated. Twenty-five percent of all children (P25) reached a minimum voice range of almost two octaves, namely, 22 semitones (ST) from 220 to 784 Hz with soft and loud singing. Half of the children (P50) had a voice range of 24 ST (2 octaves), while soft singing and a larger voice range of 26 ST while loud singing. The measurements of third quartile (P75) revealed that 25% of children have even a larger voice range than 29 dB (from 196 Hz/g to 1047 Hz/c3) and can sing at most frequencies louder than 90 dB. P90 demonstrated that 10% of the children can sing even lower or higher than the frequency range between 196 Hz/g and 1319 Hz/e3 analyzed. The voice range seems not to be constrained by social but by voice/musical background: children of vocally/musically encouraged schools had wider voice ranges. This underlines the necessity of regular singing lessons already in primary schools. The percentile VRP introduced might help to evaluate the vocal constitution and vocal capabilities of a child. PMID- 19303740 TI - Reliability and clinical validity of the Italian Reflux Symptom Index. AB - Currently, there is no Italian version of the Reflux Symptom Index (RSI). The aim of this study was to develop an Italian RSI and to evaluate its internal consistency, reliability, and clinical validity. The study design used was a cross-sectional survey study. Eighty patients with a Reflux Finding Score (RFS) >7, and 193 asymptomatic subjects were included in the study. For the RSI reliability analysis, the appositely developed Italian RSI was filled twice, with a week interval, by the 80 patients and 80 control subjects. The test-retest reliability was assessed through the Pearson correlation test, whereas the Cronbach's alpha coefficient was used for internal consistency analysis. For the clinical validity assessment, the scores obtained in the pathological group were compared with the data from the asymptomatic individuals through the Student's t test. Finally, the correlation between RSI and RFS in the 80 patients was assessed. All of the patients filled in the entire questionnaire autonomously. The test-retest reliability in the patients, as well as in the control group, was very high (r>0.90); the internal consistency also showed very high values (alpha=0.99). The mean RSI score in the patients was 21.1+/-6.6, whereas in the control group it was 6.3+/-5.6; the difference was statistically significant (P=0.0001). The mean RFS score in the 80 patients was 9.2+/-2.7 and the correlation between RFS score and RSI score was rather high (r=0.89). The Italian RSI is easily administered, highly reproducible, and exhibits excellent clinical validity. PMID- 19303741 TI - Gradation of stiffness of the mucosa inferior to the vocal fold. AB - During phonation, energy is transferred from the subglottal airflow through the air/mucosa interface that results in the propagation of the mucosal wave in the vocal fold. The vocal fold is soft, and the subglottal mucosa is stiff. We hypothesize that it is highly improbable that there is a rigid boundary between the tissue structures, with a sudden drop in stiffness; and that a gradual change would be more likely to support the efficient transfer of energy from the airflow to the mucosal wave. Our objective was to test this hypothesis by quantifying the change in mucosa stiffness with respect to anatomical position. In this initial study, using five pig larynges, a series of point-specific measurements of mucosa stiffness were taken in a line from the midpoint of the vocal fold toward the trachea. A modified linear skin rheometer adapted for laryngeal elasticity measurement applied shear stress to a series of seven positions at 2-mm intervals starting from the midmembranous vocal fold medial surface. A sinusoidal shear force of 1g was applied at each point, and resultant displacement curve logged. Using a regression algorithm, the stiffness of the tissue was derived in units of grams force per millimeter displacement. Five readings were taken at each position. The results indicate that there is a linear increase in stiffness with respect to position, increasing as the measurements are taken further from the vocal fold. There is a gradual change in stiffness of the subglottal mucosa of a pig larynx. PMID- 19303742 TI - The fate of titanium miniplates and screws used in maxillofacial surgery: a 10 year retrospective study. AB - The objective of this 10 year, retrospective study is to evaluate the indications for the removal of titanium miniplates following osteosynthesis in maxillofacial trauma and orthognathic surgery. All patients who had miniplates placed in a Regional Oral and Maxillofacial Department between January 1998 and October 2007 were included. The following variables were recorded: patient gender and age, number of plates inserted, indications for plate placement, location of plates, number and location of plates removed, indications for plate removal, time between insertion and removal, medical co-morbidities, and the follow-up period. During the 10 years of the study, 1247 titanium miniplates were placed in 535 patients. A total of 32 (3%) plates were removed from 30 patients. Superficial infection accounted for 41% of all plates removed. All complications were minor and most plates were removed within the first year of insertion. A low removal rate of 3% suggests that the routine removal of asymptomatic titanium miniplates is not indicated. PMID- 19303743 TI - Addition of verapamil in the treatment of severe myoclonic epilepsy in infancy. AB - We report on the use of the voltage-gated calcium channel blocker (Vg-CCB), verapamil, as an add-on anticonvulsant medication in two girls, 4 and 14 years of age, who were affected by severe myoclonic epilepsy in infancy (SMEI) or Dravet syndrome, a channelopathy caused by abnormalities in the voltage-gated sodium channel neuronal type alpha1 subunit (SCN1A) gene at 2q24. Both girls had pharmacoresistant epilepsy and developmental delay. Mutation analysis for the SCN1A gene revealed a missense mutation in exon 2 in the 4-year-old girl. Verapamil was co-administered in both children with a prompt response in controlling status epilepticus, myoclonic jerks, and partial and generalized seizures. The therapeutic effect lasted 13 months in the 14-year-old girl, while it is still present after a 20-month follow-up period in the 4-year-old girl who, in addition, has experienced improvement in motor and language development. The verapamil vVg-CCB, which crosses the blood-brain barrier (BBB): (a) inhibits the P-glycoprotein, an active efflux transporter protein expressed in normal tissue, including the brain, which is believed to contribute to the in situ phenomenon of multidrug resistance; and (b) may regulate membrane depolarization induced by abnormal sodium channels functions by modulating the abnormal Ca++ influxes into neurons with subsequent cell resting. This is the first report on long-lasting verapamil therapy in SMEI. The functional consequences of such in vivo modulating effects on Ca++ channels could contribute to rational targeting for future molecular therapeutic approaches in pharmacoresistant epileptic channelopathies. PMID- 19303744 TI - Epidemiological trends in psychosis-related Emergency Department visits in the United States, 1992-2001. AB - Mental health visits represented an increasing fraction of all Emergency Department (ED) visits in the U.S. between 1992 and 2001. This study used the National Hospital Ambulatory Medical Care Survey, a 4-staged probability sample of ED visits from geographically diverse hospitals around the U.S., to assess the contribution of all psychosis-related visits to this overall trend. Unlike other mental-health-related ED visits, the rate of psychosis-related visits did not increase. This lack of change is notable in the context of dramatic changes in both healthcare financing and antipsychotic prescribing practices during this period. There was an unexpected decrease in Medicare-funded psychosis-related ED visits at a time of increasing Medicare enrollment overall. An important demographic trend over this decade was the increasing urbanization of psychosis related ED visits coincident with a relative decrement in such visits within rural areas. PMID- 19303745 TI - Topical calcineurin inhibitors in atopic dermatitis: a systematic review and meta analysis. AB - OBJECTIVES: To build a critical appraisal of the available literature to evaluate the effectiveness of topical calcineurin inhibitors in treatment of atopic dermatitis (AD), in comparison to topical corticosteroids (TCs) and/or placebo. REVIEW METHODS: DESIGN: systematic review and meta-analysis. DATA SOURCES: electronic search of MEDLINE Pubmed along the last 10 years (1997-2006). STUDY SELECTION: randomized control trials of TCIs reporting efficacy outcomes, in comparison to TCs or vehicle (placebo) or both. DATA SYNTHESIS: of 210 articles, 19 studies were included, 10 for tacrolimus and 9 for pimecrolimus, involving 7378 patients of whom 2771 applied tacrolimus, 1783 applied pimecrolimus, and 2824 were controls. Both drugs were significantly more effective than a vehicle. However, two long-term trials comparing demonstrated the value of pimecrolimus in reduction of flares and steroid-sparing effect after 6 months. Compared to TCs, both 0.1% and 0.03% tacrolimus ointments were as effective as moderate potency TCs, and more effective than a combined steroid regimen. Tacrolimus was more effective than mild TCs. CONCLUSIONS: TCIs in AD are more effective than placebo. Although less effective than TCs, pimecrolimus has its value in long-term maintenance and as a steroid-sparing agent in AD, whenever used early enough, at first appearance of erythema and/or itching. In treatment of moderate to severe AD, topical tacrolimus is as effective as moderately potent TCs, and more effective than mild preparations. Chronic AD lesions of the face and flexures are the most justified indication for topical calcineurin inhibitors. PMID- 19303746 TI - Blood, adipose tissue and brain levels of the cannabinoid ligands WIN-55,212 and SR-141716A after their intraperitoneal injection in mice: compound-specific and area-specific distribution within the brain. AB - Cannabinoid ligands have wide ranging neural and behavioral effects; therefore, they are of substantial therapeutic interest. The levels of cannabinoids are tightly controlled in brain infusion and in vitro methodologies, although the studied dose-ranges are extremely wide (e.g. 0.4-470 nmol in brain infusion studies). The brain levels reached after systemic administration are virtually unknown. To investigate this issue, we injected intraperitoneally (3)H-labeled WIN-55,212 and SR141716A (0.3, 1 and 3 mg/kg) and estimated their accumulation in the blood, adipose tissue and brain. Accumulation was dose-dependent. The largest amounts were found in the adipose tissue, while the levels seen in the blood and brain were approximately similar. The accumulation of SR141716A was markedly more pronounced than that of WIN-55,212 in all three tissues. The brain distribution of WIN-55,212 showed large regional differences. Such differences were significant but much smaller with SR141716A. The largest brain levels noticed after intraperitoneal injections did not exceed 2.5 nmol/g. This is larger than the brain level of the endocannabinoid anandamide but smaller than that of 2 arachidonoyl glycerol. Yet, the CB1 receptor affinity of WIN-55,212 and SR 141716A is two orders of magnitude larger than that of 2-arachidonoyl glycerol, suggesting that the exogenously administered compounds were functionally more active. Our findings also suggest that brain infusion and in vitro techniques employing considerably larger doses than 2.5 nmol should be dealt with caution. It appears that measuring brain levels after systemic injections increases our understanding of cannabinoid effects, and provides important clues for the comparison of results obtained with different methodologies. PMID- 19303747 TI - The effect of surface microtopography of poly(dimethylsiloxane) on protein adsorption, platelet and cell adhesion. AB - Chemical homogeneous poly(dimethylsiloxane) (PDMS) surface with dot-like protrusion pattern was used to investigate the individual effect of surface microtopography on protein adsorption and subsequent biological responses. Fibrinogen (Fg) and fibronectin (Fn) were chosen as model proteins due to their effect on platelet and cell adhesion, respectively. Fg labeled with (125)I and fluorescein isothiocyanate (FITC) was used to study its adsorption on flat and patterned surfaces. Patterned surface has a 46% increase in the adsorption of Fg when compared with flat surface. However, the surface area of the patterned surface was only 8% larger than that of the flat surface. Therefore, the increase in the surface area was not the only factor responsible for the increase in protein adsorption. Clear fluorescent pattern was visualized on patterned surface, indicating that adsorbed Fg regularly distributed and adsorbed most on the flanks and valleys of the protrusions. Such distribution and local enrichment of Fg presumably caused the specific location of platelets adhered from platelet rich plasma (PRP) and flowing whole blood (FWB) on patterned surface. Furthermore, the different combination of surface topography and pre-adsorbed Fn could influence the adhesion of L929 cells. The flat surface with pre-adsorbed Fn was the optimum substrate while the virgin patterned surface was the poor substrate in terms of L929 cells spread. PMID- 19303748 TI - The morphology and post-hatching development of the skull of Bolitoglossa nicefori (Caudata: Plethodontidae): developmental implications of recapitulation and repatterning. AB - The cranial morphology of the direct-developing salamander Bolitoglossa nicefori and its post-hatching development are described and compared with that of other urodeles. Four stages of cranial development are defined on the basis of conspicuous events that occur during post-hatching ontogeny. The adult skull morphology of B. nicefori is similar to that of other plethodontids; however, some regions show interspecific variation. The post-hatching ontogeny of the skull and the stage of ossification observed in the hatchlings of B. nicefori show two important ontogenetic features: (1) a mosaic of early larval, metamorphic and post-metamorphic skull features in hatchlings, and (2) absence of characteristic larval elements in skull and hyoid apparatus. The distinctive stage of ossification in the hatchlings of B. nicefori could be caused by heterochronic changes in the ossification sequence, compared to the ontogeny of metamorphic salamanders. The possible heterochronic changes and the absence of larval traits are perhaps due to ontogenetic repatterning, yet without an obvious impact on the adult skull morphology (absence of morphological novelties). This might indicate a compartmentalized development. Further studies should be performed in order to establish the possible occurrence of recapitulatory patterns or ontogenetic repatterning in the skull morphogenesis of B. nicefori during its embryonic development. PMID- 19303749 TI - Silymarin BIO-C, an extract from Silybum marianum fruits, induces hyperprolactinemia in intact female rats. AB - Breastfeeding is widely acknowledged to have important health benefits for infants and mothers. Milk thistle (Silybum marianum fruits) has been recently proposed to be used by nursing mothers for stimulating milk production; however, the mode of action of this herbal drug is still unknown. In this paper, we have evaluated the effect of a micronized standardized extract of S. marianum (Silymarin BIO-C=Piulatte) on the serum levels of prolactin in female rats. A 14 day treatment with Silymarin BIO-C (25-200mg/kg, given orally) increased, in a dose dependent manner, the serum prolactin levels. Moreover, after a 66-day discontinuation of Silymarin BIO-C treatment, prolactin levels were still significantly elevated although we observed a trend to decrease that was counteracted by a further 7-day treatment with Silymarin BIO-C. Bromocriptine, a dopamine D(2) receptor agonist, (1-10mg/kg, os) significantly and in a dose dependent manner, reduced the serum prolactin levels; bromocriptine, at the dose of 1mg/kg, significantly reduced the high serum prolactin levels induced by Silymarin BIO-C. In conclusion, we have shown that an extract from S. marianum fruits significantly increases circulating prolactin levels in female rats; this effect seems to involve, at least in part, dopamine D(2) receptors. PMID- 19303750 TI - In vitro immunostimulatory properties of Abrus lectins derived peptides in tumor bearing mice. AB - In vitro immunostimulatory effect of Abrus lectins derived peptide fractions (AGP and ABP) was investigated in DL bearing mice. Both AGP and ABP were found to activate splenocytes and induced production of cytokines like IL-2, IFN-gamma and TNF-alpha indicating a Th1 type of immune response. Analysis of in vitro treated splenocytes by flow cytometry revealed an increase in percentage of T and B cell with high expression of activation markers (CD25(+) and CD71(+)). At the same time, expression of co-stimulatory markers was significantly high compared to tumor control. The tumor associated macrophages were able to stimulate NO production, IL-1 secretion, increased phagocytosis and decreased expression of mannose receptor. It was also observed that NK cell was activated by AGP and ABP. These results suggest that both AGP and ABP act as immunostimulants in vitro in DL bearing mice. PMID- 19303752 TI - Nuphar lutea: in vitro anti-leishmanial activity against Leishmania major promastigotes and amastigotes. AB - Several anti-leishmanial drugs of choice are of plant origin. Many of the available drugs against the disease are toxic and in certain cases parasite drug resistance is developed. The development of new compounds is urgently required. AIMS OF THE STUDY: To determine the leishmanicidal activity of the Nuphar lutea plant extract against Leishmania major in vitro. MATERIALS AND METHODS: The leishmanicidal activity of methanolic plant extract against L. major free living promastigotes and intracellular amastigotes was evaluated, using microscopic examinations and the enzymatic XTT assay. RESULTS: Methanolic extract of N. lutea was highly effective against both Leishmania promastigotes and L. amastigotes (IC(50)=2+/-0.12 microg/ml; ID(50)=0.65+/-0.02 3 microg/ml; LD(50)=2.1+/-0.096 microg/ml, STI=3.23). The extract at 1.25 microg/ml totally eliminated the intracellular parasites within 3 days of treatment. Also, a synergistic anti leishmanial activity was demonstrated with N. lutea extract combined with the anti-leishmanial drug--paromomycin. The partially purified N. lutea active component was found to be a thermo-stable alkaloid(s) with no electrical charge and is resistant to boiling and to methanol, dichloromethane and xylene treatment. CONCLUSIONS: The present study suggests that N. lutea might be a potential source of anti-leishmanial compounds. PMID- 19303751 TI - Hypoglycemic activity of a novel anthocyanin-rich formulation from lowbush blueberry, Vaccinium angustifolium Aiton. AB - Blueberry fruits are known as a rich source of anthocyanin components. In this study we demonstrate that anthocyanins from blueberry have the potency to alleviate symptoms of hyperglycemia in diabetic C57b1/6J mice. The anti-diabetic activity of different anthocyanin-related extracts was evaluated using the pharmaceutically acceptable self-microemulsifying drug delivery system: Labrasol. Treatment by gavage (500 mg/kg body wt) with a phenolic-rich extract and an anthocyanin-enriched fraction formulated with Labrasol lowered elevated blood glucose levels by 33 and 51%, respectively. The hypoglycemic activities of these formulae were comparable to that of the known anti-diabetic drug metformin (27% at 300 mg/kg). The extracts were not significantly hypoglycemic when administered without Labrasol, demonstrating its bio-enhancing effect, most likely due to increasing the bioavailability of the administered preparations. The phenolic rich extract contained 287.0+/-9.7 mg/g anthocyanins, while the anthocyanin enriched fraction contained 595+/-20.0 mg/g (cyanidin-3-glucoside equivalents), as measured by HPLC and pH differential analysis methods. The greater hypoglycemic activity of the anthocyanin-enriched fraction compared to the initial phenolic-rich extract suggested that the activity was due to the anthocyanin components. Treatment by gavage (300 mg/kg) with the pure anthocyanins, delphinidin-3-O-glucoside and malvidin-3-O-glucoside, formulated with Labrasol, showed that malvidin-3-O-glucoside was significantly hypoglycemic while delphinidin-3-O-glucoside was not. PMID- 19303753 TI - Berberine inhibits human tongue squamous carcinoma cancer tumor growth in a murine xenograft model. AB - Our primary studies showed that berberine induced apoptosis in human tongue cancer SCC-4 cells in vitro. But there is no report to show berberine inhibited SCC-4 cancer cells in vivo on a murine xenograft animal model. SCC-4 tumor cells were implanted into mice and groups of mice were treated with vehicle, berberine (10mg/kg of body weight) and doxorubicin (4mg/kg of body weight). The tested agents were injected once per four days intraperitoneally (i.p.), with treatment starting 4 weeks prior to cells inoculation. Treatment with 4mg/kg of doxorubicin or with 10mg/kg of berberine resulted in a reduction in tumor incidence. Tumor size in xenograft mice treated with 10mg/kg berberine was significantly smaller than that in the control group. Our findings indicated that berbeirne inhibits tumor growth in a xenograft animal model. Therefore, berberine may represent a tongue cancer preventive agent and can be used in clinic. PMID- 19303754 TI - Modulation of in vitro murine B-lymphocyte response by curcumin. AB - Curcumin is a phenolic natural product isolated from the rhizome of Curcuma longa (tumeric). It was previously described that curcumin had a potent anti inflammatory effect and inhibited the proliferation of a variety of tumor cells. In the present study, we investigated the inhibitory effects of curcumin on the response of normal murine splenic B cells. Curcumin inhibited the proliferative response of purified splenic B cells from BALB/c mice stimulated with the Toll like receptor ligands LPS and CpG oligodeoxynucleotides. LPS-induced IgM secretion was also inhibited by curcumin. The proliferative response induced by either the T-independent type 2 stimuli anti-delta-dextran or anti-IgM antibodies was relatively resistant to the effect of curcumin. We investigated the intracellular signaling events involved in the inhibitory effects of curcumin on murine B cells. Curcumin did not inhibit the increase in calcium levels induced by anti-IgM antibody. Western blotting analysis showed that curcumin inhibited TLR ligands and anti-IgM-induced phosphorylation of ERK, IkappaB and p38. Curcumin also decreased the nuclear levels of NFkappaB. Our results suggested that curcumin is an important inhibitor of signaling pathways activated upon B cell stimulation by TLR ligands. These data indicate that curcumin could be a potent pharmacological inhibitor of B cell activation. PMID- 19303755 TI - Nitric oxide inhibitory principles from Derris trifoliata stems. AB - Nine rotenoids were isolated from the hexane and dichloromethane extracts of Derris trifoliata stems and were tested for nitric oxide (NO) inhibitory activity using RAW264.7 cells. The result indicated that 12a-hydroxyrotenone (7) possessed very potent NO inhibitory activity with an IC(50) value of 0.002 microM, followed by 1 (deguelin, IC(50)=0.008 microM), 9 (12a-hydroxyelliptone, IC(50)=0.010 microM) and 2 (alpha-toxicarol, IC(50)=0.013 microM), respectively. In addition, the DPPH scavenging activity of rotenoids was also investigated. It was found that 6a,12a-dehydrodeguelin (5) possessed the highest activity against DPPH with an IC(50) value of 7.4 microM, followed by deguelin (1, IC(50)=27.4 microM). All compounds did not show any cytotoxicity at their IC(50) values for NO inhibitory activity. Structure-activity relationships (SARs) of these rotenoids against NO release are as follows: (1) hydroxylation at C12a dramatically increased activity, (2) prenylation at furan ring increased activity markedly and (3) hydrogenation of a double bond at C6a-C12a conferred higher activity. For the DPPH radical scavenging effect, it was found that (1) introduction of a double bond at C6a-C12a increased activity and (2) hydroxylation of C11 at the D-ring decreased activity. As regards active compounds of Derris trifoliata stems, the isolated compounds are responsible for the NO inhibitory effect, especially 7, 1, 9 and 2, whereas 5 and 1 are those for the DPPH scavenging activity. PMID- 19303756 TI - Alcohol extract of Echinacea pallida reverses stress-delayed wound healing in mice. AB - Healing of open skin wounds begins with an inflammatory response. Restraint stress has been well documented to delay wound closure, partially via glucocorticoid (GC)-mediated immunosuppression of inflammation. Echinacea, a popular herbal immunomodulator, is purported to be beneficial for wound healing. To test the hypothesis, an alcohol extract of E. pallida was administrated orally to mice for 3 days prior to, and 4 days post wounding with a dermal biopsy on the dorsum. Concomitantly, mice were exposed to 3 cycles of daily restraint stress prior to, and 4 cycles post wounding. Echinacea accelerated wound closure in the stressed mice, but had no apparent wound healing effect for the non-stressed mice when compared to their respective controls. To test if the positive healing effect is through modulation of GC release, plasma corticosterone concentrations were measured in unwounded mice treated with restraint stress and the herbal extract for 4 days. Plasma GC in restraint stressed mice gavaged with Echinacea was not different from mice treated with restraint only, but was increased compared to the vehicle control. This data suggests that the improved wound healing effect of Echinacea in stressed mice is not mediated through modulation of GC signaling. PMID- 19303757 TI - Rat electropharmacograms of the flavonoids rutin and quercetin in comparison to those of moclobemide and clinically used reference drugs suggest antidepressive and/or neuroprotective action. AB - In order to be able to test single constituents of herbal plant extracts with respect to possible clinical usefulness, the model of local field potential analysis leading to the so-called electropharmacogram has been successfully used in rats to classify the effects of theanine and theogallin in the past. The present investigation aims at the prediction of efficacy and possible mechanisms of action of rutin and quercetin. Adult rats (day-night converted) were instrumented with four bipolar concentric electrodes into the frontal cortex, hippocampus, striatum and reticular formation. Field potentials were recorded during a pre-drug reference period of 45 min followed by oral administration of the particular test compound and 4h recording thereafter. Data were transmitted wirelessly to the computer for spectral frequency analysis. Rutin (5-80 mg/kg) as well as quercetin (5-40 mg/kg orally) produced similar electropharmacograms with dose dependent decreases of spectral alpha2 and beta1 frequencies within all brain areas. Peak effects were reached 4h after administration. The pattern of changes approached that obtained after 2.5mg/kg of moclobemide during the first hour as revealed by discriminant analysis in comparison to a large matrix of other drugs with known clinical indications. Data suggest antidepressant capabilities for rutin and quercetin with inhibition of monoamino oxidase at least as part of the mechanism of action. Both compounds should be tested clinically in patients with symptoms of depression. PMID- 19303759 TI - RNA silencing in germlines--exquisite collaboration of Argonaute proteins with small RNAs for germline survival. AB - As the proper development of germlines is vital for species preservation, elaborative, regulatory systems for gene expression must operate in germlines. One such system is RNA silencing, sequence-specific gene silencing mechanisms mediated by small RNAs of 20-30 nucleotides long. Indeed, recent studies have revealed that various types of small RNAs are expressed germline-specifically. To preserve the germlines, they collaborate with Argonaute proteins, the catalytic engines in RNA silencing, to inhibit injurious, parasitic genes, transcriptionally or post-transcriptionally. This chapter summarizes the exquisite collaboration of Argonaute proteins with small RNAs in the RNA silencing mechanisms necessary for germline survival in Drosophila and mice. PMID- 19303758 TI - PIKKing on PKB: regulation of PKB activity by phosphorylation. AB - Ser/Thr protein kinase PKB/Akt is a key regulator of a wide range of cellular processes including growth, proliferation and survival. PKB is clearly a crucial signaling molecule and extensive research efforts aim to understand its regulation and action. Recent studies of the regulation of PKB activity by hydrophobic motif phosphorylation have yielded several exciting findings about members of the PI3-kinase-like family of kinases (PIKKs) acting as PKB regulators. Mammalian target of rapamycin complex 2 (mTORC2) and DNA-dependent protein kinase (DNA-PK) can both phosphorylate Ser473 and activate PKB. This present review concerns PKB regulation by mTORC2 and DNA-PK in a stimulus dependent and context-dependent manner and the possible implications of this for PKB activity, substrate specificity and therapeutic intervention. PMID- 19303760 TI - The legacy of 'normalisation': the role of classical and contemporary criminological theory in understanding young people's drug use. AB - Since it began in the mid-1990s, the debate surrounding the normalisation of adolescent recreational drug use has attracted considerable attention and has tended to polarise opinion within the field. In this article two of the main protagonists in the debate come together to discuss its legacy. Focusing on the twin themes of continuity and change the authors begin by considering the relevance of early developments in the sociology of drug use, noting that this earlier work anticipated much that has recently been written on the subject, including the emphasis on hedonism and consumption in leisure lifestyles. From here they go on to critically reflect on the role that structure and agency have played in the normalisation debate, suggesting that the original thesis underplayed the role of structural influences in favour of a rational action model of adolescent drug use. In their more recent work, both authors have come to emphasise how drug use is shaped by an interplay between social structure and human agency. While some areas of disagreement remain, they agree that normalisation is best understood as a contingent process negotiated by distinct social groups operating in bounded situations. PMID- 19303761 TI - Cross-national comparison of adolescent drinking and cannabis use in the United States, Canada, and the Netherlands. AB - BACKGROUND: This research examined the prevalence of drinking and cannabis use among adolescents in the United States, Canada, and the Netherlands, countries with substantially different laws and policies relating to these substances. METHODS: Laws regarding drinking and cannabis use were rated for each country. Substance use prevalence data among 10th graders from the Health Behaviour in School-Aged Children Survey conducted in each country in 2005-2006 were examined. RESULTS: Laws regarding alcohol and cannabis were found to be strictest in the United States, somewhat less strict in Canada, and least strict in the Netherlands. On most measures of drinking, rates were lower in the United States than in Canada or the Netherlands. With United States as the referent, relative risks (RR) for monthly drinking were 1.30 (1.11-1.53) for Canadian boys and 1.55 (1.31-1.83) for girls, and 2.0 (1.73-2.31) for Dutch boys and 1.92 (1.62-2.27) for Dutch girls. Drunkenness was also higher among Canadian boys and girls and Dutch boys. However, rates of cannabis use did not differ between the countries, except that Dutch girls were less likely to use cannabis in the past year (RR=.67; .46-.96). CONCLUSIONS: The lower prevalence of adolescent drinking and drunkenness (except among Dutch girls) in the United States is consistent with the contention that strict drinking policies may limit drinking among 10th graders. However, the finding that cannabis use rates did not differ across countries is not consistent with the contention that prohibition-oriented policies deter use or that liberal cannabis policies are associated with elevated adolescent use. Based on these findings, the case for strict laws and policies is considerably weaker for cannabis than for alcohol. PMID- 19303762 TI - Seasonal characterization of municipal solid waste (MSW) in the city of Chihuahua, Mexico. AB - Management of municipal solid waste (MSW) has become a significant environmental problem, especially in fast-growing cities. The amount of waste generated increases each year and this makes it difficult to create solutions which due to the increase in waste generation year after year and having to identify a solution that will have minimum impact on the environment. To determine the most sustainable waste management strategy for Chihuahua, it is first necessary to identify the nature and composition of the city's urban waste. The MSW composition varied considerably depending on many factors, the time of year is one of them. Therefore, as part of our attempt to implement an integral waste management system in the city of Chihuahua, we conducted a study of the characteristics of MSW composition for the different seasons. This paper analyzes and compares the findings of the study of the characterization and the generation of solid waste from households at three different socio-economic levels in the city over three periods (April and August, 2006 and January, 2007). The average weight of waste generated in Chihuahua, taking into account all three seasons, was 0.592 kg capita(-1) day(-1). Our results show that the lowest income groups generated the least amount of waste. We also found that less waste was generated during the winter season. The breakdown for the composition of the waste shows that organic waste accounts for the largest proportion (45%), followed by paper (17%) and others (16%). PMID- 19303763 TI - Comparing microbial die-off in separately collected faeces with ash and sawdust additives. AB - In a urine diversion dry toilet (UDDT), the urine and faeces are collected separately in order to recycle their nutrient content unmixed. In a UDDT, an additive e.g. lime, wood ash, dry soil or sawdust, depending on which one is easily accessed by the users, is usually sprinkled to the faeces after each defecation. The purpose of the additive is primarily to keep away the flies and odours and to contribute to primary treatment of the faeces. In this paper, ash and sawdust were applied separately to source-separated faeces during the collection phase, and then the die-off of indicators and pathogens in the mixtures was studied. The die-off of E. coli in the faeces/ash mixture was faster initially (first 7 days) compared to that achieved in the faeces/sawdust mixture even though the die-off achieved after 30-50 days was nearly similar for both mixtures. E. coli was not detected in faeces/ash after about 2 months, but was detected after 2 months in the faeces/sawdust mixture. Enterococcus spp. did not decrease below detection in faeces/ash or faeces/sawdust mixture but higher numbers (difference of about 2 logs) were detected at all times in faeces/sawdust than in faeces/ash mixture. The difference in the die-off in the mixtures of faeces/ash and faeces/sawdust was attributed to the differences in the characteristics of the additives, namely, high alkaline mineral content (giving high pH) and lower moisture content of ash compared to sawdust. It is recommended to increase use of ash as additive over sawdust in urine diversion dry toilets. PMID- 19303764 TI - Perspectives for genetic engineering for the phytoremediation of arsenic contaminated environments: from imagination to reality? AB - Phytoremediation to clean up arsenic-contaminated environments has been widely hailed as environmentally friendly and cost effective, and genetic engineering is believed to improve the efficiency and versatility of phytoremediation. Successful genetic engineering requires the thorough understanding of the mechanisms involved in arsenic tolerance and accumulation by natural plant species. Key mechanisms include arsenate reduction, arsenic sequestration in vacuoles of root or shoot, arsenic loading to the xylem, and volatilization through the leaves. Key advances include the identification of arsenic (As) translocation from root to shoot in the As hyperaccumulator, Pteris vittata, and the characterization of related key genes from hyperaccumulator and nonaccumulators. In this paper we have proposed three pathways for genetic engineering: arsenic sequestration in the root, hyperaccumulation of arsenic in aboveground tissues, and phytovolatilization. PMID- 19303765 TI - The nuclear envelope--a scaffold for silencing? AB - An increasing number of studies indicate that chromosomes are spatially organized in the interphase nucleus and that some genes tend to occupy characteristic zones of the nuclear volume. FISH studies in mammalian cells suggest a differential localization of active and inactive loci, with inactive heterochromatin being largely perinuclear. Recent genome-wide mapping techniques confirm that the nuclear lamina, which lies beneath the nuclear envelope, interacts preferentially with silent genes. To address the functional significance of spatial compartmentation, gain-of-function assays in which chromatin is targeted to the nuclear periphery have now been carried out. Such experiments yielded coherent models in yeast; however, conflicting results in mammalian cells leave it unclear whether these concepts apply to higher organisms. Nevertheless, the recent discovery that evolutionarily conserved inner nuclear membrane proteins support the peripheral anchoring of yeast heterochromatin suggests that certain principles of nuclear organization may hold true from yeast to man. PMID- 19303766 TI - Spatial representation across species: geometry, language, and maps. AB - We review growing evidence that the reorientation system-shared by both humans and nonhuman species-privileges geometric representations of space and exhibits many of the characteristic features of modular systems. We also review evidence showing that humans can move beyond the limits of nonhuman species by using two cultural constructions, language and explicit maps. We argue that, although both of these constructions are uniquely human means of enriching the spatial system we share with other species, their representational formats, functions, and developmental trajectories are quite different, yielding distinctly different tools for empowering human spatial cognition.The capacity to reorient using geometry is present in humans by the age of 18 months. PMID- 19303768 TI - Treatment of advanced hepatocellular carcinoma with long-acting octreotide: a phase III multicentre, randomised, double blind placebo-controlled study. AB - BACKGROUND: A previous study reported a significant survival benefit for octreotide compared with no treatment in patients with advanced hepatocellular carcinoma (HCC). This was investigated further in this multicentre study. PATIENTS AND METHODS: Two hundred and seventy two patients with HCC who were ineligible for curative treatments or had relapsed following potentially curative therapies were randomised to receive long-acting octreotide, 30 mg as an intramuscular injection once every 4 weeks for up to 2 years, or placebo. RESULTS: At the time of the final analysis, median overall survival (OS) was 6.53 months (95% confidence interval [CI], 4.8-8.3) for octreotide versus 7.03 months (95% CI, 5.43-8.53) for placebo (p=0.34). Progression-free survival (p=0.26) also did not differ significantly between the two treatment groups. No objective responses were achieved in the octreotide group but 33% of patients achieved disease stabilisation for a mean time of 5.5 months (95% CI, 1.1-9.9). The median time until definitive global health score deterioration (according to QLQ-C30) was 2.3 months (95% CI, 1.4-3.7) in the octreotide and 4 months (95% CI, 2.2-5.7) in the placebo group (p=0.09). There were four objective responses in the placebo group. Octreotide was well tolerated; seven patients reported severe adverse events possibly related to octreotide and there were no cases of haematoma or cholecystitis. CONCLUSIONS: In patients with advanced HCC, octreotide has a favourable safety profile but does not improve OS and could have a negative impact on quality of life. PMID- 19303767 TI - The long and the short of riboswitches. AB - Regulatory mRNA elements or riboswitches specifically control the expression of a large number of genes in response to various cellular metabolites. The basis for selectivity of regulation is programmed in the evolutionarily conserved metabolite-sensing regions of riboswitches, which display a plethora of sequence and structural variants. Recent X-ray structures of two distinct SAM riboswitches and the sensing domains of the Mg(2+), lysine, and FMN riboswitches have uncovered novel recognition principles and provided molecular details underlying the exquisite specificity of metabolite binding by RNA. These and earlier structures constitute the majority of widespread riboswitch classes and, together with riboswitch folding studies, improve our understanding of the mechanistic principles involved in riboswitch-mediated gene expression control. PMID- 19303769 TI - Fed-batch fermentation of Tuber melanosporum for the hyperproduction of mycelia and bioactive Tuber polysaccharides. AB - For the first time, a fed-batch fermentation process of Tuber melanosporum was developed for the efficient production of bioactive mycelia and Tuber polysaccharides. Each 1.67 g/L of peptone and 8.33 g/L of yeast extract were added on day 3, 6, and 9, respectively, and sucrose was fed to maintain its concentration around 35-5 g/L when its residual level decreased to 10-5 g/L. Then, the maximal biomass, the production of extracellular polysaccharides (EPS) and intracellular polysaccharides (IPS) reached 53.72+/-2.57 g DW/L, 7.09+/-0.62 and 4.43+/-0.21 g/L, respectively. Compared with the batch culture conducted in the enriched medium, the biomass, the production of EPS and IPS were enhanced by 55.8%, 222.3% and 103.2%, respectively. Not only the cell density but also the production of EPS and IPS were the highest ever reported in truffle fermentation, and the biomass was also the highest as ever reported in mushroom fermentation. PMID- 19303770 TI - Fractionation of proteins and carbohydrates of extracellular polymeric substances in a membrane bioreactor system. AB - The major operational problem associated with membrane bioreactors (MBR) is membrane fouling, for which extracellular polymeric substances (EPS) are primarily responsible. In this work both the soluble and bound EPS (i.e. SMP and EPS) produced in an MBR system operating under sludge retention times (SRT) of 10, 15, 20 and 33 days were fractionized by means of membranes having variable molecular weight cutoffs (300 kDa, 100 kDa, 10 kDa & 1 kDa). The results show that increasing the SRT leads to a reduction of SMP and EPS and that these reductions are more pronounced for the SRTs in the range 10-20 days. This reduction is more significant for carbohydrates than for proteins. The decrease of SMP and EPS with increasing SRT from 10 to 20 days led to a significant decrease of the level of fouling. The further increase of SRT to 33 days did not significantly impact on the level of fouling as the SMP and EPS concentrations did not change much. Under the examined operating conditions, EPS were found to be composed mainly of large macromolecules having a size of 0.45 microm-300 kDa and to a lower extent of very small molecules (<1 kDa) that are not easily decomposed by the biomass activity. The majority of SMP is composed of very small molecules (<1 kDa), while some macromolecules in the range of 0.45 microm-300 kDa are present. Consequently, both EPS and SMP were found to have a bimodal character. PMID- 19303771 TI - Comparative study of lignins isolated from Alfa grass (Stipa tenacissima L.). AB - Soda lignin, dioxane lignin and milled lignin were isolated from Alfa grass (Stipatenacissima L.). The physico-chemical characterization of three different lignins: one industrial lignin precipitated from soda spent liquor and two lignin preparations isolated under laboratory conditions from Alfa grass (also know as Esparto grass) was performed. The structures of lignins were studied by three non destructive (FT-IR, solid state (13)C NMR and UV/visible spectroscopy) and two destructive (nitrobenzene oxidation and thermogravimetric analysis) methods. Elemental analysis and the methoxyl content determination were performed in order to determine the C(9) formulae for the studied lignins. The total antioxidant capacity of the studied lignins has been determined and compared to commercial antioxidants commonly used in thermoplastic industry. PMID- 19303772 TI - Changes in the bacterial community structure in two-stage constructed wetlands with different plants for industrial wastewater treatment. AB - This study focused on the diversity of bacterial communities from two series of two-stage constructed wetlands (CWs) treating tannery wastewater, under different hydraulic conditions. Series were separately planted with Typha latifolia and Phragmites australis in expanded clay aggregates and operated for 31 months. The effect of plant species, hydraulic loading and unit stage on bacterial communities was addressed through bacterial enumeration and denaturating gradient gel electrophoresis (DGGE). Diverse and distinct bacterial communities were found in each system unit, which was related in part to the type of plant and stage position (first or second unit in the series). Numerical analysis of DGGE profiles showed high diversity in each unit with an even distribution of species. No clear relation was established between the sample collection time, hydraulic loading applied and the bacterial diversity. Isolates retrieved from plant roots and substrates of CWs were affiliated with gamma-Proteobacteria, Firmicutes, alpha-Proteobacteria, Sphingobacteria, Actinobacteria and Bacteroidetes. Both series were effective in removing organic matter from the inlet wastewater, however, based on batch degradation experiments it seems that biodegradation was limited by the recalcitrant properties of the wastewater. PMID- 19303773 TI - Enzymatic activation of hydrophobic self-immolative dendrimers: the effect of reporters with ionizable functional groups. AB - Self-immolative dendrimers are uniquely structured molecules that release multiple tail units through a chain fragmentation initiated by a single cleavage at the dendrimer's core. Although bioactivation of self-immolative dendritic molecules with only two reporter groups was demonstrated, enzymatic activation failed for self-immolative dendrimers with more reporters. These large and hydrophobic dendrimers aggregated under aqueous conditions and enzyme did not efficiently trigger chain fragmentation. Here we demonstrate a simple solution to the problem of enzymatic activation of hydrophobic self-immolative dendrimers. The reporter units on the dendritic platform were equipped with ionizable functional group. Polar interactions with water significantly decreased hydrophobicity of the dendrimers and prevented aggregate formation. Consequently, hydrophobic self-immolative dendrons were effectively activated. PMID- 19303774 TI - 1-Aryl-3,4-dihydroisoquinoline inhibitors of JNK3. AB - A series of 1-aryl-3,4-dihydroisoquinoline inhibitors of JNK3 are described. Compounds 20 and 24 are the most potent inhibitors (pIC50 7.3 and 6.9, respectively in a radiometric filter binding assay), with 10- and 1000-fold selectivity over JNK2 and JNK1, respectively, and selectivity within the wider mitogen-activated protein kinase (MAPK) family against p38alpha and ERK2. X-ray crystallography of 16 reveals a highly unusual binding mode where an H-bond acceptor interaction with the hinge region is made by a chloro substituent. PMID- 19303776 TI - A new radiological approach to cyclic mastalgia: venous doppler ultrasound. AB - OBJECTIVE: To evaluate the role of the venous system in cyclical mastalgia. MATERIALS AND METHODS: 25 healthy women and 39 women with cyclic mastalgia underwent venous Doppler examination in both pre- and postmenstrual period. Upper outer quadrant veins of breast were evaluated. The impedance indices of both groups were compared in both the premenstrual and postmenstrual period. RESULTS: During the premenstrual period, venous impedance indices were significantly higher in the patients with cyclic mastalgia (p=0.004) than in the control group. After the menstrual period, there was no difference in venous impedance indices between the groups. CONCLUSION: Excessive tissue oedema might be responsible for both a tension-mediated pain and a compression of the venous system thereby showing higher impedance than that in control group. PMID- 19303775 TI - Late-onset seropositive Isaacs' syndrome after Guillain-Barre syndrome. AB - Acquired neuromyotonia, or Isaacs' syndrome, has been described in combination with a variety of other autoimmune disorders; however there has never been a report of seropositive Isaacs' syndrome in a patient with a history of Guillain Barre syndrome (GBS). Both conditions involve antibody-mediated autoimmune effects on the peripheral nervous system, although the clinical manifestations are quite different. We present a man who experienced an episode of GBS at the age of 21 and subsequently developed Isaacs' syndrome at the age of 24 which was positive for anti-voltage-gated potassium channel (VGKC) antibodies. When treated with intravenous immunoglobulins (IVIg) he developed an eczematous rash that differed markedly in pattern and duration from the usual presentation for this IVIg reaction. PMID- 19303777 TI - A novel operative technique to manage a symptomatic synovial cyst associated with an os odontoideum. AB - Synovial cysts in association with an os odontoideum are rare. All previous cases have been treated via a posterior approach with good results. The authors present a patient with a synovial cyst in association with an os odontoideum and discuss a planned two-stage procedure to excise the cyst via a neuronavigation-assisted transoral-transpharyngeal approach, followed by a posterior C1/2 fixation. PMID- 19303778 TI - Posterior projecting carotid bifurcation aneurysms. AB - Posteriorly projecting carotid bifurcation aneurysms are rare. Proximal anterior cerebral artery aneurysms are even more uncommon. Distinguishing between true posteriorly projecting carotid bifurcation aneurysms and carotid A1 aneurysms can be difficult but it is important because small posteriorly projecting carotid bifurcation aneurysms may need a contralateral approach for adequate exposure. We report our experience with a recent case that illustrated this problem. PMID- 19303779 TI - Long-term use of temozolomide: could you use temozolomide safely for life in gliomas? AB - Temozolomide (TMZ) is an alkylating agent used in the management of gliomas. Although TMZ is generally safe and acute toxicity is well documented, there are limited data on long-term toxicities. We present three patients with glioma; all patients started on TMZ after having progressed following primary treatment. These patients have continued TMZ for 5 years, 7 years and 8 years respectively. So far they have had no serious side effects. We discuss these patients while raising the question of prolonged TMZ use. PMID- 19303780 TI - Hearing preservation after stereotactic radiosurgery for vestibular schwannoma: a systematic review. AB - Radiosurgery has evolved into an effective alternative to microsurgical resection in the treatment of patients with vestibular schwannoma. We performed a systematic analysis of the literature in English on the radiosurgical treatment of vestibular schwannoma patients. A total of 254 published studies reported assessable and quantifiable outcome data of patients undergoing radiosurgery for vestibular schwannomas. American Association of Otolaryngology-Head and Neck Surgery (AAO-HNS) class A or B and Gardner-Robertson (GR) classification I or II were defined as having preserved hearing. A total of 5825 patients (74 articles) met our inclusion criteria. Practitioners who delivered an average dose of 12.5 Gy as the marginal dose reported having a higher hearing preservation rate (12.5 Gy=59% vs. >12.5 Gy=53%, p=0.0285). Age of the patient was not a significant prognostic factor for hearing preservation rates (<65 years=58% vs. >65 years=62%; p=0.4317). The average overall follow-up was 41.2 months. Our data suggest that an overall hearing preservation rate of about 57% can be expected after radiosurgical treatment, and patients treated with 12.5 Gy were more likely to have preserved hearing. PMID- 19303781 TI - Modification of the butenyl-spinosyns utilizing cross-metathesis. AB - The discovery of a strain of Saccharopolyspora sp. that produced a number of spinosyn analogs that had not before been seen gave an ideal opportunity for extending our knowledge of that SAR of these highly efficacious insecticides. In particular, these compounds contained a butenyl group connected to C-21 which in the regular spinosyns was substituted with a simple ethyl group. The double bond therefore gave us a handle to further modify this position allowing us to substitute different groups there. In this paper we show one of our approaches to this modification using olefin cross-metathesis. Even though the spinosyns were not highly efficient substrates for metathesis reactions, we were nevertheless successful in extending their chemistry accordingly. PMID- 19303782 TI - Phenolic compounds with radical scavenging and cyclooxygenase-2 (COX-2) inhibitory activities from Dioscorea opposita. AB - Phytochemical studies of the chloroform soluble fraction of Dioscorea opposita resulted in the isolation of four new compounds, 3,5-dihydroxy-4-methoxybibenzyl (1), 3,3',5-trihydroxy-2'-methoxybibenzyl (2), 10,11-dihydro-dibenz[b,f]oxepin 2,4-diol (3), and 10,11-dihydro-4-methoxy-dibenz[b,f]oxepin-2-ol (4), together with an additional fifteen known compounds. The structures of 1-4 were elucidated by spectroscopic methods including 2D NMR. All of the nineteen isolated compounds were tested in the DPPH, superoxide anion radical scavenging assays and cyclooxygenases (COXs) inhibition assay. Of those, compounds 7, 9, 11, 12, 13, 15 and 18 exhibited radical scavenging activities and compounds 2, 3, 8, 13, 15 and 16 showed selective inhibitory activities against COX-2. PMID- 19303783 TI - Experimental and theoretical determination of the low-loss electron energy loss spectroscopy of LiMn2O4. AB - The dielectric properties of cubic spinel-type LiMn(2)O(4), used as cathode material in lithium ion secondary batteries, are studied by analyzing the low loss region of the electron energy loss spectroscopy (EELS) spectrum in a transmission electron microscope. A comparison of experimental EELS spectra and ab initio density-functional theory calculations (WIEN2k code) within the generalized gradient approximation (GGA) is presented. The origins of interband transitions are identified in the electronic band structure, by calculating the partial imaginary part of the dielectric function and the partial density of states of Li, Mn and O. Good agreement with experimental spectra is observed which allowed interpreting main features of the EELS spectrum. PMID- 19303784 TI - Evaluation of coronary endothelial dysfunction in healthy young smokers: Cold pressor test using [(15)O]H(2)O PET. AB - The purpose of this study was to investigate coronary endothelial dysfunction in young healthy smokers by measuring myocardial blood flow (MBF) using [(15)O]H(2)O PET. The study population was 18 young male volunteers consisted of 9 smokers (age: 23.8+/-1.1yr) and 9 non-smokers (age: 25.0+/-2.5yr). The smokers had been smoking cigarettes for 6.6+/-2.5 pack years. Myocardial [(15)O]H(2)O-PET was performed at rest, during cold (5 degrees C) pressor stimulation and during adenosine infusion. Left ventricular (LV) input function and tissue time-activity curves were obtained by drawing region of interest (ROI) on the LV blood pool and myocardium images obtained by non-negative matrix factorization (NMF) of dynamic [(15)O]H(2)O-PET data, and MBF was calculated using these time-activity curves and single compartmental model. There were no significant difference in resting MBF between two groups (smokers: 1.43+/-0.41 and non-smokers: 1.37+/ 0.41ml/g/min; P=NS). However, during cold pressor stimulation, MBF in smokers was significantly lower than that in non-smokers (1.25+/-0.33 vs. 1.59+/ 0.29ml/g/min; P=0.019). MBF changed to 90+/-24% of resting MBF in smokers and 122+/-28% in non-smokers. The difference in the ratio of cold pressor MBF to basal MBF between two groups was also significant (P=0.024). During adenosine infusion, however, hyperemic MBF did not differ significantly between smokers and non-smokers (5.81+/-1.99 vs. 5.03+/-1.27ml/g/min; P=NS). This study shows that [(15)O]H(2)O PET analysis can reveal that endothelial dysfunction occurs in even young smokers of about 6 pack years. PMID- 19303785 TI - Three-dimensional dosimetry in brachytherapy: A MAGAT study. AB - This study is to evaluate the influence of using different matrix size of smoothing filter for image post-processing and various slice thickness during MR imaging on dose estimation in Ir-192 HDR brachytherapy via normoxic polymer gel dosimeter. Our results show its sensitive nature in gel dosimeter while changing these parameters, among which the combination of 2mm slice thickness of MR images and [5x5] smoothing filter are considered the optimal parameters to provide accurate dose estimations and isodose curves. PMID- 19303786 TI - New concept of CZT-based hand-held radioisotope identifier. AB - Radiation dose can be determined with the radiation dosimeter. Recently, hand held radioisotope identifier is being developed by many researchers to measure the radiation dose and to determine the type of the radioisotope. New design concept of the radioisotope identifier is introduced in this work. In this design, one can measure the radiation dose and the type of the radioisotope, and one can also determine the direction where the radiation comes. The hand-held radioisotope identifier is based on cadmium zinc telluride (CZT) detector. The detection principle is from the change of the energy spectrum as the direction of the incident radiation is varied. EGSnrc Monte Carlo simulation is done to calculate the energy spectrum of the CZT detector, and the result is compared with the measured one. The energy spectra are calculated when the gamma-ray is incident on the detection at different directions. The simulation results show the change of the spectrum shape as the incident radiation comes from different directions. The calculation is also done with different radiation energies and detector sizes. A prototype radioisotope identifier is being developed at Korea Atomic Energy Research Institute (KAERI). This work would be helpful to develop next generation radiation dosimeter. PMID- 19303787 TI - Development of scaling factors for the activated concrete of the KRR-2. AB - The biological shielding concrete of KRR-2 was activated by a thermal neutron reaction during the operation of the reactor, thus a variety of radionuclides were generated in the concrete. In order to verify the radioactivity for the final disposal of waste and to achieve a more efficient cutting of the concrete, the radioactivity inventories and distributions of the activated concrete were evaluated. The activity of gamma-emitting radionuclides was measured by using an HPGe detector. The beta-emitting radionuclides were measured by an oxidation/combustion method for (3)H and (14)C and a combined method of an extraction chromatography and a liquid scintillation for (55)Fe and (63)Ni. The dominant radioactive nuclides in the activated concrete were (3)H, (14)C, (55)Fe and (60)Co, and the maximum gamma activity was 105Bq/g at the surface around the thermal column. The specific activities of all the nuclides were found to decrease almost linearly on a logarithmic scale along the depth from the inner surface of the concrete. Equations for scaling factors were obtained by a linear regression of logarithms from the radioactivity data of (3)H/(60)Co, (14)C/(60)Co and (55)Fe/(60)Co nuclide pairs of the activated concrete. The scaling factors can be utilized for the estimation of beta radioactivity without the time consuming separation processes of the nuclides. PMID- 19303788 TI - Determination of dispersion coefficients using radioisotope data in river environment. AB - A field tracer experiment using radioisotope (82)Br was performed to estimate the dispersion characteristics of pollutants in river environment. The dispersion coefficients in the longitudinal and transverse directions were determined by using the measured concentration of a radioisotope. Numerical models were applied to calculate the flow and concentration fields at the experimental site. Several numerical simulations were performed to investigate the effects of the numerical results according to variations of the dispersion coefficients. The calculated results for several runs were compared with the measured ones by using statistical methods. The calculated concentrations agreed well with the measured ones. PMID- 19303789 TI - Differential expression of alpha-l-arabinofuranosidase and alpha-l arabinofuranosidase/beta-d-xylosidase genes during peach growth and ripening. AB - Arabinose is the major neutral sugar in peach (Prunus persica (L.) Batsch) cell walls and substantial changes in arabinose content take place not only during peach melting, when a rapid-softening-related depolymerizing activity may be expected, but also at the onset of peach ripening. A full-length cDNA clone sequence referred to as PpARF1 (GenBank accession no. DQ486870) was obtained and determined by bioinformatics' analysis to be a peach alpha-l-arabinofuranosidase homologue. The deduced PpARF1 translation product is 677 amino acids in length while the mature protein has a predicted molecular mass of 71.6 kD and a theoretical pI of 4.94. Semi-quantitative RT-PCR reactions were conducted to evaluate the expression of both PpARF1 and PpARF/XYL (GenBank accession no. AB264280), the latter encoding a putative bifunctional protein displaying both alpha-l-arabinofuranosidase and beta-d-xylosidase activities. In peach fruit, the PpARF1 gene expression was detected at every developmental stage with a maximum during S2 (lag phase of development) and a subsequent decrease towards S4 (maximal fruit size). In contrast, PpARF/XYL transcript levels were relatively high at the end of S1 (fruit set) and at S3-E (beginning of the cell expansion). Substantial increases in PpARF1 mRNA levels were found at the beginning and end of the climacteric rise and also in melting fruit. In contrast, PpARF/XYL transcripts reached a maximum when fruit firmness was 22-26 N, with a slight decline during the melting stage. PpARF/XYL and PpARF1 were expressed differently in three fruit tissue types as well as in other plant tissues. Ethylene is regarded as the main regulator of peach ripening and the accumulation of PpARF/XYL and PpARF1 transcripts is coincident with the autocatalytic ethylene production during ripening. On the hand, other factors may also play a role in PpARF1 and PpARF/XYL expression, since transcripts accumulate at different developmental times and organs even when ethylene biosynthesis is barely detectable. PMID- 19303790 TI - Acquired von Willebrand syndrome after exchange of the HeartMate XVE to the HeartMate II ventricular assist device. AB - Instead of pulsatile ventricular assist devices an increasing number of nonpulsatile ventricular assist devices are introduced to clinical practice. The different flow characteristics of this new technique lead to alteration in shear stress on blood components, which may affect the coagulation system. Repeated von Willebrand factor analyses were performed in a patient who first was implanted with a pulsatile ventricular assist device (Thoratec HeartMate XVE), which had to be replaced after 405 days with an axial flow device (HeartMate II). During support with the pulsatile ventricular assist device there was no sign of any coagulation disorder. However, on the axial flow device acquired von Willebrand syndrome Type 2 developed. Inhibition of platelet function was also observed, which may be in part due to the von Willebrand syndrome. The HeartMate II axial flow device may induce von Willebrand syndrome, which was not observed in HeartMate XVE pulsatile ventricular assist device. Patients put on continuous flow devices should be screened for acquired von Willebrand syndrome. PMID- 19303791 TI - Seventeen years of adult congenital heart surgery: a single centre experience. AB - OBJECTIVE: With a growing number of children with congenital heart disease (CHD) reaching adulthood, an extensive experience with cardiac surgery in adults with CHD is accumulating. To increase insight in this patient category we report our 17-year single centre experience including predictors for adverse outcome and EuroSCORE performance. METHODS: Patients and operative characteristics of all consecutive adult CHD patients operated upon between January 1990 and January 2007 were collected. Categorisation was done according to the EACTS/STS congenital database. Early and late morbidity and mortality were assessed with follow-up extending up to 17 years. EuroSCORE performance was assessed. RESULTS: Nine hundred and sixty-three procedures were performed in 830 patients (mean age 39.3 years, 50.3% male). A total of 49% were re-do procedures, frequent procedures were for left heart lesions (37%), right heart lesions (31%) and septal defects (8%). The 51% primary procedures largely consisted of less complex procedures but also included 1.4% of tetralogy of Fallot repairs, 4.1% of aortic coarctation repairs and 2.7% of Ebstein's disease repairs. Thirty-day mortality was 1.5% (n=14); predicted mortality by logistic EuroSCORE was 4.6%. c-index was 0.61 (95% CI 0.46-0.75). Major complications such as tamponade requiring intervention occurred in 3.2%, postoperative bleeding requiring re-exploration in 7.1% and renal insufficiency requiring dialysis in 4 (0.4%). Pulmonary hypertension was a strong predictor for short-term mortality; impaired ventricular function and cyanosis for long-term mortality. Overall 17-year survival was 71% (95% CI 61%-82%). Eighty percent of patients were in NYHA class I at last follow-up, 17% in II, 3% in III, 0% in IV. CONCLUSIONS: Surgery in adult CHD patients can be performed with low operative mortality and good clinical outcome. EuroSCORE is not a good model for risk assessment in this group of patients. PMID- 19303792 TI - Pulmonary trunk perforation during transapical minimal invasive aortic valve replacement. AB - In this case report we illustrate our experience with a perforation of the pulmonary trunk during a transapical aortic valve implantation in a single case. The patient suffered from an aortic valve stenosis and was accepted for a minimally invasive procedure because of multiple comorbidities. After unproblematic transapical placement of the aortic valve, a venous bleeding from the anterolateral wound was observed. Median sternotomy showed a bleeding out of the pulmonary trunk that could be stopped with purse-string sutures. The further course of the patient was uneventful. PMID- 19303793 TI - Thyroid function during coronary surgery with and without cardiopulmonary bypass. AB - OBJECTIVE: Cardiopulmonary bypass (CPB) is associated with thyroid hormone changes consistent with euthyroid sick syndrome. Similar changes have been observed after general surgical operations. Thyroid hormone changes and their association with global oxygen consumption were studied in low-risk patients undergoing coronary artery bypass grafting (CABG) with and without CPB. METHODS: Fifty-two patients undergoing primary CABG by the same surgeon were randomised into either on-pump (ONCAB, n=26) or off-pump (OPCAB, n=26) groups. Thyroid stimulating hormone (TSH), free thyroxine (fT4) and free triiodothyronine (fT3) levels were measured at sequential time-points using chemiluminescence assays. Global oxygen consumption was measured at sequential time-points using a continuous cardiac output Swan-Ganz catheter. RESULTS: In both groups TSH and fT4 remained within normal range throughout the study. There was a similar and progressive decline in fT3 levels with no significant difference between the groups over time (p=0.42). Mean fT3 levels at 24h were below the normal range and significantly lower than baseline values (ONCAB, 3.3+/-0.69 pmol/L vs 5.1+/-0.41 pmol/L, p<0.001; OPCAB, 3.3+/-0.51 pmol/L vs 5.0+/-0.46 pmol/L, p<0.001). There was a significant inverse relationship between fT3 levels and global oxygen consumption. CONCLUSIONS: Off-pump surgery is associated with thyroid hormone changes similar to conventional surgical revascularisation. The data suggest that further studies into T3 administration during OPCAB may be warranted. PMID- 19303794 TI - Therapeutic decisions for patients with symptomatic severe aortic stenosis: room for improvement? AB - OBJECTIVE: Symptomatic severe aortic stenosis is an indication for aortic valve replacement. Some patients are denied intervention. This study provides insight into the proportion of conservatively treated patients and into the reasons why conservative treatment is chosen. METHODS: Of a patient cohort presenting with severe aortic stenosis between 2004 and 2007, medical records were retrospectively analyzed. Only symptomatic patients (n=179) were included. We studied their characteristics, treatment decisions, and survival. RESULTS: Mean age was 71 years, 50% were male. During follow-up (mean 17 months, 99% complete) 76 (42%) patients were scheduled for surgical treatment (63 conventional valve replacement, 10 transcatheter, 1 heart transplantation, 2 waiting list) versus 101 (56%) who received medical treatment. Reasons for medical treatment were: perceived high operative risk (34%), symptoms regarded mild (19%), stenosis perceived non-severe (14%), and patient preference (9%). In 5% the decision was pending at the time of the analysis and in 20% the reason was other/unclear. Mean age of the surgical group was 68 years versus 73 years for medically treated patients (p=0.004). Predicted mortality (EuroSCORE) was 7.8% versus 11.3% (p=0.006). During follow-up 12 patients died in the surgical group (no 30-day operative mortality), versus 28 in the medical group. Two-year survival was 90% versus 69%. CONCLUSIONS: A large proportion (56%) of symptomatic patients does not undergo aortic valve replacement. Often operative risk is estimated (too) high or hemodynamic severity and symptomatic status are misclassified. Interdisciplinary team discussions between cardiologists and surgeons should be encouraged to optimize patient selection for surgery. PMID- 19303795 TI - Methyl group migration during the fragmentation of singly charged ions of trimethyllysine-containing peptides: precaution of using MS/MS of singly charged ions for interrogating peptide methylation. AB - Core histones are susceptible to a range of post-translational modifications (PTMs), including acetylation, phosphorylation, methylation, and ubiquitination, which play important roles in the epigenetic control of gene expression. Here, we observed an unusual discrepancy between MALDI-MS/MS and ESI-MS/MS on the methylation of trimethyllysine-containing peptides with residues 9-17 from human histone H3 and residues 73-83 from yeast histone H3. It turned out that the discrepancy could be attributed to an unusual methyl group migration from the side chain of trimethyllysine to the C-terminal arginine residue during peptide fragmentation, and this methyl group transfer only occurred for singly charged ions, but not for doubly charged ions. The methyl group transfer argument received its support from the results on the studies of the fragmentation of the ESI- or MALDI-produced singly charged ions of several synthetic trimethyllysine bearing peptides. The results presented in this study highlighted that caution should be exerted while MS/MS of singly charged ions is employed to interrogate the PTMs of trimethyllysine-containing peptides. PMID- 19303796 TI - The influence of increasing steady-state walking speed on muscle activity in below-knee amputees. AB - The goal of this study was to identify changes in muscle activity in below-knee amputees in response to increasing steady-state walking speeds. Bilateral electromyographic (EMG) data were collected from 14 amputee and 10 non-amputee subjects during four overground walking speeds from eight intact leg and five residual leg muscles. Using integrated EMG measures, we tested three hypotheses for each muscle: (1) there would be no difference in muscle activity between the residual and intact legs, (2) there would be no difference in muscle activity between the intact leg and non-amputee legs, and (3) muscle activity in the residual and intact legs would increase with speed. Most amputee EMG patterns were similar between legs and increased in magnitude with speed. Differences occurred in the residual leg biceps femoris long head, vastus lateralis and rectus femoris, which increased in magnitude during braking compared to the intact leg. These adaptations were consistent with the need for additional body support and forward propulsion in the absence of the plantar flexors. With the exception of the intact leg gluteus medius, all intact leg muscles exhibited similar EMG patterns compared to the control leg. Finally, the residual, intact and control leg EMG all had a significant speed effect that increased with speed with the exception of the gluteus medius. PMID- 19303797 TI - Reducing the NMR sample volume using a single organic liquid: increased sensitivity for mass-limited samples with standard NMR probes. AB - A simple inexpensive protocol for confining an aqueous sample to the active region of a standard NMR probe is examined for high-resolution NMR. The aqueous sample is sandwiched between an inert perfluorinated organic liquid that has been exploited in the design of micro-coil NMR probes. The procedure is demonstrated with 3mm NMR tubes at ambient and elevated temperatures but should be equally applicable to smaller diameter tubes. It is shown that confinement has minimal effects on line shape and provides at least a two fold increase in sensitivity over a conventional sample, for the same mass of solute. PMID- 19303798 TI - Double quantum transition as the origin of the central dip in the z-spectrum of HDO in variably stretched gel. AB - The 2H NMR spectrum of HDO in gelatin gel, that is stretched inside silicone rubber tubing, displays a well resolved doublet. Spectra were obtained with a range of offset frequencies of partially saturating radio-frequency (RF) radiation. The resulting steady-state irradiation envelope (also referred to as a 'z-spectrum') has the peculiar feature that maximal suppression of the doublet occurs when the irradiation is applied exactly at the centre frequency, between the two HDO peaks. We present a quantum mechanical explanation for this phenomenon. It is shown that the phenomenon is the result of double quantum transitions. The analysis is extendable to more complex quadrupolar and dipolar coupled systems of other nuclides. It has implications for enhancement of contrast in magnetic resonance imaging of heterogeneous systems using dipolar and quadrupolar interactions. PMID- 19303799 TI - Projection of a quantum state on eigenstates of average Hamiltonian. AB - A scheme for projecting an arbitrary quantum state on eigenstates of average Hamiltonian is described. As an experimental example, projection on entangled Bell states, which are eigenstates of specially constructed average Hamiltonian, is demonstrated for a system of two dipolar-coupled nuclear spins. The results of a direct and time-reversed evolution are added to average out the coherences between different eigenstates and accomplish the projection. PMID- 19303800 TI - The First International Standard For Insulin-like Growth Factor-1 (IGF-1) for immunoassay: preparation and calibration in an international collaborative study. AB - The World Health Organization (WHO) Expert Committee on Biological Standardization (ECBS) has recognized the need for an International Standard for Insulin-like Growth Factor-1 (IGF-1) for the calibration of immunoassays and for the monitoring of the content of therapeutic products. The objective of the study reported here was the characterization of a candidate standard for IGF-1 in an international collaborative study carried out by 18 laboratories in nine countries, by comparison with (i) a primary calibrant characterized by amino acid analysis and UV spectroscopy, and (ii) the existing International Reference Reagent coded 87/518 by HPLC, immunoassay and bioassay. The study was designed as follows: Phase I involved the establishment of a primary calibrant of rhIGF-1, containing approximately 1.0mg rhIGF-1 per vial. A defined value was assigned to the primary calibrant by amino acid analysis (AAA) and UV spectroscopy. Phase II involved calibration of the candidate standard in terms of the primary calibrant by HPLC, with confirmatory data from immunoassay and bioassay. Results from Phase I confirmed the primary calibrant as containing 1.045 mg per vial. Although there was some variability among laboratory estimates of IGF-1 in the proposed standard using the different methods in Phase II, the estimates by the various methods were in broad agreement. On the basis of the results reported here, the World Health Organization (WHO) has established the preparation coded 02/254 as the First International Standard for Insulin-like Growth Factor-1, human, recombinant, for immunoassay with an assigned content of 8.50 microg per ampoule. Details of how to order the standard can be found at www.nibsc.ac.uk. PMID- 19303801 TI - On the effect of thalidomide on Mycobacterium avium subspecies paratuberculosis in culture. AB - BACKGROUND: Without known mechanisms of action, thalidomide is used to treat a variety of non-malignant 'idiopathic' diseases. There is increasing concern that Mycobacterium avium subspecies paratuberculosis (MAP) may be zoonotic. Recently, methotrexate, azathioprine, 6-mercaptopurine (6-MP), 5-aminosalicylic acid (5 ASA), cyclosporine A, rapamycin, and tacrolimus have been shown to inhibit MAP growth in culture, indicating that, unknowingly, MAP infections may have been treated for decades. We herein test the hypothesis that thalidomide may inhibit MAP growth. METHODS: Using the radiometric 14CO2 (Bactec) system we quantified growth kinetics of thalidomide (+/-), (+), and (-) and two components for thalidomide, phthalimide and 1-hydroxypiperidine-2,6-dione (HPD). We studied four MAP strains (three human isolates, 'Ben', 'Dominic', and UCF-4, and a bovine MAP isolate 19698) and three mycobacterial controls (Mycobacterium avium and bacillus Calmette-Guerin (BCG)). Growth was quantified as growth index (GI) and inhibition as percent decrease in cumulative GI (%-DeltacGI). RESULTS: Phthalimide had no dose-dependent inhibition on any strain. Neither thalidomide nor HPD inhibited M. avium or BCG. MAP inhibition varied; at 64 microg/ml, amongst human isolates, Dominic was most susceptible: thalidomide (+)=58%-DeltacGI and HPD=46%-DeltacGI. UCF-4 was next: thalidomide (-)=37%-DeltacGI and HPD=40%-DeltacGI. Ben was least susceptible: HPD=24%-DeltacGI. CONCLUSIONS: We have shown, in culture, the heretofore-undescribed inhibition of MAP growth by thalidomide and its enantiomers. Phthalimide was found to have no anti-MAP activity, whereas HPD was found to inhibit MAP growth. These data are compatible with the hypothesis that thalidomide, like other 'anti-inflammatories' and 'immunomodulators' may act, in part, as an anti-MAP antibiotic. PMID- 19303802 TI - Prevalence of subclinical amyloidosis in Tunisian patients with rheumatoid arthritis. AB - INTRODUCTION: Secondary amyloidosis is a serious complication of rheumatoid arthritis (RA). Symptoms are late to occur, so that screening is in order, most notably in patients with long-standing RA. The objectives of our study were to determine the prevalence of subclinical amyloidosis in RA patients by abdominal fat aspiration biopsy (AFAB) and minor salivary gland biopsy (MSGB) and to identify factors associated with subclinical amyloidosis. METHODS: We prospectively studied 107 consecutive patients with RA (94 women and 13 men) recruited between March 2005 and January 2006. Clinical and laboratory findings, imaging study results, and treatment were recorded for each patient. AFAB and MSGB were performed routinely. Amyloid deposits were identified by polarized light microscopy after Congo red staining. RESULTS: The prevalence of subclinical amyloidosis was 21.5% by AFAB and 3.7% by MSGB. Factors associated with subclinical amyloidosis were a longer time to diagnosis (P=0.03), extraarticular manifestations (P=0.019), proteinuria >0.3 g/24 h (P=0.024), and absence of methotrexate therapy (P=0.046). Subclinical amyloidosis was not associated with age, sex, RA duration, joint deformities, DAS28 score, Health Assessment Questionnaire score, Steinbrocker radiological stage, rheumatoid factor, erythrocyte sedimentation rate, C-reactive protein, creatinine, or hemoglobin. CONCLUSION: The prevalence of subclinical amyloidosis by AFAB is high (21.5%). AFAB is more sensitive than MSGB for detecting subclinical amyloidosis. A simple screening tool such as AFAB should be used, particularly in patients with risk factors. Subclinical amyloidosis requires close monitoring to ensure the early detection and treatment of symptomatic amyloidosis. PMID- 19303803 TI - Use of short-term bio-impedance for gastric motility assessment. AB - Gastric motility (GM) has been assessed using bio-impedance (BI) techniques in the time domain using short term recordings and analyzing mainly the median of the area under the fast Fourier transform (FFT) spectra. The BI technique has been applied to the gastric system mainly for gastric emptying (GE) studies. However, gastric motility evaluation using BI has not been fully implemented. In this study, we propose the use of the BI technique for the evaluation of gastric motility considering global features of the fast FFT spectra. The study was performed in eleven healthy subjects who were evaluated in fasting and postprandial conditions. The results indicate that the use of the median of the area under the FFT spectra is equivalent to the use of the main peak of the spectra to determine the changes in gastric motility from the fasting to the postprandial state. This demonstrates that BI is a valid technique for gastric motility evaluation in short time recordings. PMID- 19303804 TI - Peripheral myoclonus and muscle hypertrophy of quadriceps revealing a spinal dysraphism. PMID- 19303805 TI - Towards a teleoperated needle driver robot with haptic feedback for RFA of breast tumors under continuous MRI. AB - OBJECTIVE: The purpose of this paper is to explore the feasibility of developing a MRI-compatible needle driver system for radiofrequency ablation (RFA) of breast tumors under continuous MRI imaging while being teleoperated by a haptic feedback device from outside the scanning room. The developed needle driver prototype was designed and tested for both tumor targeting capability as well as RFA. METHODS: The single degree-of-freedom (DOF) prototype was interfaced with a PHANToM haptic device controlled from outside the scanning room. Experiments were performed to demonstrate MRI-compatibility and position control accuracy with hydraulic actuation, along with an experiment to determine the PHANToM's ability to guide the RFA tool to a tumor nodule within a phantom breast tissue model while continuously imaging within the MRI and receiving force feedback from the RFA tool. RESULTS: Hydraulic actuation is shown to be a feasible actuation technique for operation in an MRI environment. The design is MRI-compatible in all aspects except for force sensing in the directions perpendicular to the direction of motion. Experiments confirm that the user is able to detect healthy vs. cancerous tissue in a phantom model when provided with both visual (imaging) feedback and haptic feedback. CONCLUSION: The teleoperated 1-DOF needle driver system presented in this paper demonstrates the feasibility of implementing a MRI compatible robot for RFA of breast tumors with haptic feedback capability. PMID- 19303806 TI - Daptomycin: mechanisms of action and resistance, and biosynthetic engineering. AB - Daptomycin is a lipopeptide antibiotic used clinically to treat infections caused by Gram-positive bacteria. Laboratory studies have shown that Staphylococcus aureus resistance to daptomycin occurs stepwise and slowly. Mutations associated with decreased susceptibility were mapped in mprF, yycG, rpoB, and rpoC, each giving about twofold increases in the minimal inhibitory concentration (MIC) and combinations giving higher MICs. The mprF gene encodes a dual functional enzyme that couples lysine to phosphatidylglycerol (PG) and transfers the lysyl-PG (LPG) to the outer leaflet of the membrane. LPG is less acidic than PG, and thus reduces the binding of Ca(++)-bound daptomycin to bacterial membranes. The mprF mutants have higher LPG/PG ratios in the membrane outer leaflet and bind less daptomycin than the wild-type strain. YycG is a sensor histidine kinase of a two component signal transduction system required for viability in many low G+C Gram positive bacteria. The observation of DapR mutations in yycG suggests that YycG may be a target for daptomycin antibacterial activity. Daptomycin inserts into PG rich membrane at the cell division septum, but also inserts into lung surfactant, explaining why it failed to meet non-inferiority criteria in clinical trials for community acquired pneumonia (CAP). Recent advances in biosynthetic engineering have provided new tools to generate novel lipopeptides with modifications in the core peptide: several were very potent antibiotics against Gram-positive pathogens, and some were active in the presence of surfactant. PMID- 19303807 TI - Synthesis of 1-phenyl-3-biphenyl-5-(N-ethylcarbazole-3-yl)-2-pyrazoline and its use as DNA probe. AB - A novel pyrazoline derivative, named 1-phenyl-3-biphenyl-5-(N-ethylcarbazole-3 yl)-2-pyrazoline, was synthesized, and its structure was confirmed by means of IR, (1)H NMR, and elementary analysis. The compound emits strong yellow fluorescence. Decrease of fluorescence intensity of the pyrazoline derivative in the presence of calf thymus DNA (ct DNA) is observed, and the quenching obey Stern-Volmer equation. There is a single quenching mechanism for the complex, which belongs to static quenching. KI quenching study shows that the magnitude of K(SV) of the bound pyrazoline is lower than that of the free one. It is also found that ionic strength could affect the interaction. Binding constants for pyrazoline with ct DNA and salmon sperm DNA (ss DNA) are in the same order of 10(4) molL(-1), and binding site size are about 1 per base pairs. Experimental results indicate that the new compound might insert into DNA base pairs by intercalative binding mode. PMID- 19303808 TI - Detection of Epstein-Barr virus DNA in peripheral blood is associated with the development of bronchiolitis obliterans syndrome after lung transplantation. AB - BACKGROUND: The long-term success of lung transplantation is limited by the development of bronchiolitis obliterans syndrome (BOS). Virus infections may be involved in the development of BOS. OBJECTIVES: The study intended to investigate whether there is an association of Epstein-Barr virus (EBV), cytomegalovirus (CMV) and human adenovirus (HAdV) with the development of BOS and to identify risk factors for EBV detection in blood. STUDY DESIGN: A prospective cohort study was conducted in lung and heart-lung transplant recipients (LTR) who are followed in our outpatient clinic. 385 LTR were monitored for CMV pp65 antigen, EBV and HAdV DNA in blood at follow-up visits for 6 months. The development of BOS was recorded for a median of 21 months. RESULTS: EBV DNA, HAdV DNA and CMV pp65 antigen were detected at least once in, respectively, 202/385 LTR (52.5%), 10/382 LTR (2.6%) and 19/385 LTR (4.9%). Repeated EBV DNA detection and acute rejection were associated with the development of BOS. Variables associated with EBV DNA detection in blood were the diagnosis of BOS before study entry, retransplantation and immunosuppressive therapy with sirolimus or everolimus. CONCLUSIONS: EBV reactivation is frequent in LTR. The variables found associated with EBV reactivation probably reflect increased immunosuppression. Repeated EBV DNA detection in blood, possibly reflecting chronic EBV replication, is associated with the development of BOS. The elucidation of whether and how EBV DNAemia triggers the development of BOS could improve long-term survival of LTR. PMID- 19303809 TI - Laser evoked potential recording from intracerebral deep electrodes. AB - OBJECTIVE: To investigate whether recording from deep intracerebral (IC) electrodes can disclose laser evoked potential (LEP) components generated under the cerebral cortex. METHODS: LEPs were recorded to hand and/or perioral region stimulation from 7 patients suffering from Parkinson's disease, who underwent implant of IC electrodes in the globus pallidum pars interna (GPi), in the subthalamic nucleus (STN) and in the pedunculopontine nucleus (PPN). LEPs were obtained from the IC electrode contacts and from the Cz vertex, referred to the nose. RESULTS: The scalp traces showed a triphasic response (P1-N2-P2). The IC electrodes recorded two main components (ICP2 and ICN2), showing the same latencies as the scalp N2 and P2 potentials, respectively. The ICP2-ICN2 complex was sometimes preceded by a ICP1 wave at the same latency of the scalp P1 response. CONCLUSIONS: The LEP components recorded from the IC electrodes mirrored the ones picked up from the Cz lead, thus suggesting that they are probably generated by the opposite pole of the same cortical sources producing the scalp responses. SIGNIFICANCE: In the IC traces, there was no evidence of earlier potentials possibly generated within the thalamus or of subcortical far field responses. This means that the nociceptive signal amplification occurring within the cerebral cortex is necessary to produce identifiable LEP components. PMID- 19303810 TI - Selective modulation of intracortical inhibition by low-intensity Theta Burst Stimulation. AB - OBJECTIVE: Theta Burst Stimulation (TBS) is a repetitive transcranial magnetic stimulation paradigm which has effects on both excitatory and inhibitory intracortical pathways when applied at an intensity of 80% of active motor threshold. As intracortical inhibitory pathways have a lower threshold for activation than excitatory pathways, we sought to determine whether it was possible to selectively target cortical inhibitory circuitry by reducing the intensity of TBS to 70% of active motor threshold. METHODS: Motor evoked potentials (MEPs), short latency intracortical facilitation (SICF), intracortical facilitation (ICF) and short interval intracortical inhibition (SICI) were measured at baseline, 5-20 and 20-35 min following continuous (cTBS) and intermittent (iTBS) low-intensity TBS in nine healthy subjects. RESULTS: Low intensity cTBS significantly reduced SICI 5-20 min following stimulation, whilst having no effect on MEPs, SICF or ICF. Low-intensity iTBS had no effect on SICI, MEPs, SICF or ICF. CONCLUSIONS: It is possible to selectively target intracortical inhibitory networks for modulation by low-intensity TBS, however, responses may critically depend upon the particular paradigm chosen. SIGNIFICANCE: These findings have important implications for the treatment of neurological disorders where abnormal levels of intracortical inhibition are present, such as Parkinson's disease and focal hand dystonia and requires further investigation. PMID- 19303811 TI - Combined adaptive servo-ventilation and automatic positive airway pressure (anticyclic modulated ventilation) in co-existing obstructive and central sleep apnea syndrome and periodic breathing. AB - BACKGROUND: The co-existence of obstructive and central sleep apnea/hypopnea syndrome (OSAS) and periodic breathing is common in patients with and without underlying heart diseases. While automatic continuous positive airway pressure (APAP) has proven to effectively treat OSAS, the adaptive servo-ventilation (ASV) sufficiently improves periodic breathing. This is the first trial on a device which combines both treatment modes. METHODS: Pilot study on a two-week treatment in patients with co-existing obstructive and central and periodic breathing disturbances during sleep. Twelve consecutive patients (9 male, 3 female, age 56.9+/-10.6 years, BMI 32.4+/-5.5 kg/m(2)) were treated with a new algorithm which combines APAP and ASV (also called anticyclic modulated ventilation (ACMV), SOMNOventCR, Weinmann, Hamburg, Germany). Seven suffered from arterial hypertension, coronary heart disease and mitral regurgitation, none from congestive heart failure. RESULTS: The total apnea-hypopnea index (AHI) improved from 43.8+/-24.0/h to 2.1+/-2.4 (p<0.01), the obstructive AHI from 12.8+/-14.3/h to 0.3+/-0.6/h (p<0.01) and the central AHI from 31.0+/-17.5/h to 1.7+/-2.0/h (p<0.01). Moreover, there was a significant improvement in the total number of arousals, respiratory induced arousals, oxygen saturation and sleep profile. CONCLUSION: The algorithm combining automatic continuous positive airway pressure (CPAP) and ASV normalizes all types of co-existing obstructive and central apnea/hypopnea and periodic breathing. PMID- 19303812 TI - Does the UK really have an effective cancer plan? Revisited. PMID- 19303813 TI - Population-based cancer survival trends in England and Wales up to 2007: an assessment of the NHS cancer plan for England. AB - BACKGROUND: The National Health Service (NHS) cancer plan for England was published in 2000, with the aim of improving the survival of patients with cancer. By contrast, a formal cancer strategy was not implemented in Wales until late 2006. National data on cancer patient survival in England and Wales up to 2007 thus offer the opportunity for a first formal assessment of the cancer plan in England, by comparing survival trends in England with those in Wales before, during, and after the implementation of the plan. METHODS: We analysed population based survival in 2.2 million adults diagnosed with one of 21 common cancers in England and Wales during 1996-2006 and followed up to Dec 31, 2007. We defined three calendar periods: 1996-2000 (before the cancer plan), 2001-03 (initialisation), and 2004-06 (implementation). We estimated year-on-year trends in 1-year relative survival for patients diagnosed during each period, and changes in those trends between successive periods in England and separately in Wales. Changes between successive periods in mean survival up to 5 years after diagnosis were analysed by country and by government office region of England. Life tables for single year of age, sex, calendar year, deprivation category, and government office region were used to control for background mortality in all analyses. FINDINGS: 1-year survival in England and Wales improved for most cancers in men and women diagnosed during 1996-2006 and followed until 2007, although not all trends were significant. Annual trends were generally higher in Wales than in England during 1996-2000 and 2001-03, but higher in England than in Wales during 2004-06. 1-year survival for patients diagnosed in 2006 was over 60% for 12 of 17 cancers in men and 13 of 18 cancers in women. Differences in 3-year survival trends between England and Wales were less marked than the differences in 1-year survival. North-South differences in survival trends for the four most common cancers were not striking, but the North West region and Wales showed the smallest improvements during 2001-03 and 2004-06. INTERPRETATION: The findings indicate slightly faster improvement in 1-year survival in England than in Wales during 2004-06, whereas the opposite was true during 2001-03. This reversal of survival trends in 2001-03 and 2004-06 between England and Wales is much less obvious for 3-year survival. These different patterns of survival suggest some beneficial effect of the NHS cancer plan for England, although the data do not so far provide a definitive assessment of the effectiveness of the plan. PMID- 19303814 TI - Assessment of the NHS cancer plan in England. PMID- 19303815 TI - Was the NHS cancer plan worth the effort? PMID- 19303816 TI - Haemorrhagic cystitis and polyomavirus JC infection in ataxia telangiectasia. AB - We report the case of a young adolescent with ataxia telangiectasia (AT) and life threatening haemorrhage from the bladder due to a combination of bladder wall telangiectasis, immunosuppressive therapy and an infection with polyomavirus JC. BK and JC are both members of the polyomavirus family. BK virus is a known cause of haemorrhagic cystitis in bone-marrow and nephropathy in kidney transplant patients, whereas JC virus is mainly associated with progressive multifocal leukoencephalopathy and only rarely found in haemorrhagic cystitis. Although opportunistic infections are uncommon in AT and virus replication was described as being down-regulated in ATM (AT mutated protein)-deficient cells, clinicians should be aware that severe haematuria in a patient with AT and undergoing immunosuppressive therapy is suggestive for polyomavirus JC-induced haemorrhagic cystitis. PMID- 19303817 TI - Treatment of clitoromegaly of culturally diverse patients. AB - OBJECTIVE: Clitoroplasty is a procedure usually performed in young children, but its long-term psychosexual importance is controversial. We present two adult women from different cultural backgrounds who desired surgery for clitoromegaly. METHODS: A similar clitoroplasty procedure with preservation of the neurovascular bundle was performed on both patients. Preoperative and postoperative interviews were conducted by an independent observer to learn about the effects of the condition and the surgery. RESULTS: The Icelandic patient had bilateral ovarian dysgerminomas and a gonadoblastoma. The Bolivian patient had a 17beta hydroxysteroid oxidoreductase deficiency. Both patients reported normal libido but sexual inactivity because they felt 'embarrassed'. Postoperatively, our patients reported normal clitoral sensation and that they were sexually active. They differed in their opinion regarding the optimal timing of clitoroplasty - one suggesting surgery soon after birth and the other recommending deferral until informed consent by the patient is possible. CONCLUSIONS: Regardless of cultural background patients are affected by clitoromegaly. This is demonstrated in these cases by the reported feelings of discomfort, distress, and sexual inactivity due to embarrassment. The long-term significance of clitoromegaly and the value of clitoroplasty for young patients with disorders of sexual differentiation remain controversial. PMID- 19303818 TI - Early viraemia clearance during antiviral therapy of chronic hepatitis C improves dendritic cell functions. AB - Plasma and cellular HCV RNA and core antigen were tested in monocyte-derived DC (MDDC) from chronic hepatitis C patients undergoing treatment with peg-interferon alpha2b/ribavirin. DC allostimulatory capacity, HCV-specific T-cell reactivity and IL-12 production were measured at baseline and treatment week (TW)12. Using DC and autologous CD4(+)T-cells, obtained at baseline and TW12, we performed cross-over experiments to determine the relative role of DC and/or T-cells for impaired immune reactivity to HCV. HCV RNA and HCV core plasma levels had an impact on DC phenotype and allostimulatory capacity. In contrast, HCV genome/core protein, although detectable in DC from some patients had no effect on DC function. Antiviral immunity at TW12 was not improved in patients who remained HCV RNA positive, while early viraemia clearance (TW12) improved antiviral responses. The cross-over experiment revealed that changes in DC, rather than CD4(+)T cells have a major role for enhanced anti-HCV responses. PMID- 19303819 TI - The management of miscarriage. AB - Women diagnosed with incomplete and delayed miscarriage are faced with three options for their subsequent management: expectant, medical or surgical. Health care practitioners must empower patients to make educated decisions about their own management by providing them with sufficient information in a readily understandable format. This can be difficult both for the patient and the staff in what is often an understandably, highly emotional situation. Detailed counselling is an essential part of the process as psychological outcomes have been shown to be improved when women feel in control of the decision-making process. In this article, we discuss each of the treatment options in detail, and explore how the type of miscarriage influences their relative success rates. We also consider the comparative risks of bleeding, infection, side effects of drugs, pain scores and quality-of-life scores associated with each method through evidence derived from systematic reviews, meta-analyses and randomised controlled trials. PMID- 19303820 TI - Pigmented villonodular synovitis of a lumbar intervertebral facet joint. AB - BACKGROUND CONTEXT: Pigmented villonodular synovitis (PVNS) is a slowly progressive lesion of uncertain etiology that involves the synovial membrane of joints or tendon sheaths. Only rarely does PVNS affect the axial skeleton, where it arises from the vertebral articular facet joint. Its treatment and prognosis remains limited. PURPOSE: To describe our management in a patient with PVNS and to review previously published cases. STUDY DESIGN: Case report. METHODS: This is a case report of a 59-year-old woman who presented left sciatica. Computed tomography (CT) imaging revealed a mixed sclerotic and lucent lesion affecting the left L4-L5 facet joint. Magnetic resonance imaging (MRI) demonstrated a diffusely infiltrative process that originated from the left inferior articular process of L4 vertebra with extension into the spinal canal. A total synovectomy with left L4 hemilaminectomy was performed. Left L5 root was decompressed with total microscopic tumor removal. Decompression of spinal canal and absence of the tumor was shown by MRI and CT scan after the operation. RESULTS: Complete resolution of the patient's complaints was achieved. Histopathological analysis was consistent with a diagnosis of PVNS. CONCLUSIONS: The principle of surgical management of spinal lesions causing neurologic deficit is early surgical decompression. It is also important to totally remove the synovium, the origin of PVNS, to prevent the recurrence. PMID- 19303821 TI - Transient suppression of heart rate complexity in concussed athletes. AB - Heart rate variability (HRV) and complexity (HRC) were calculated at rest and during an isometric hand grip test (IHGT) within 48-hours (48 h) and two weeks (Week Two) of a concussion in athletes (CG) and control subjects. No differences were present at rest or in HRV during IGHT. HRC was significantly lower in the CG compared to controls at 48 h during IHGT. In CG at Week Two during IHGT, HRC was significantly greater than 48 h observations and not significantly different than controls. The findings suggest that HRC may have utility in detecting efferent cardiac autonomic anomalies within two weeks of concussion. PMID- 19303822 TI - Synthesis of phthalocyanine stabilized rhodium nanoparticles and their application in biosensing of cytochrome c. AB - A single step synthesis route is described for the preparation of rhodium nanoparticles using a cobalt aminophthalocyanine macrocyclic complex as a stabilizer. The results of nanoparticles characterization using electronic absorption, Raman and X-ray spectroscopes as well as transmission electron microscopy are reported. Rhodium nanoparticle modified electrode behavior as examined by cyclic and differential pulse voltammetry is also provided. The nanoparticles were found to be well dispersed and stabilized throughout the macromolecular matrix. TEM studies showed that they have an average diameter of 3 to 5 nm with spherical shape. The colloidal rhodium was then used for electrochemical sensing of cytochrome c using glassy carbon electrode. The results showed that the colloidal rhodium nanoparticles enhanced the electron transfer process between cytochrome c and the electrode. Differential pulse voltammetric measurements of cytochrome c at the colloidal rhodium nanoparticles modified glassy carbon electrode showed a linear relationship with the oxidation peak currents in the concentration range of 100 nM to 3 microM of cytochrome c. PMID- 19303824 TI - Racial differences in body mass indices of men imprisoned in 19th Century Texas. AB - A limited amount of research has been done on the body mass index values of 19th century Americans. This paper uses Texas prison records to demonstrate that, in contrast to today's distributions, most BMI values were in the normal range. Only 21.5% and 1.2% of the population was overweight or obese, while today comparable figures are 36% and 23%. There was also little change in BMI values between 1876 and 1919. Farmers were consistently heavier than non-farmers, while Southwestern men had lower BMI values than their counterparts from other regions of the US. BMI values indicate that 19th century African-Americans, and whites populations were well fed in spite of large expenditures on energy. PMID- 19303823 TI - Discovering novel brain lipids by liquid chromatography/tandem mass spectrometry. AB - Discovery and structural elucidation of novel brain lipids hold great promise in revealing new lipid functions in the brain and in understanding the biochemical mechanisms underlying brain physiology and pathology. The revived interests in searching for novel brain lipids have been stimulated by the expanding knowledge of the roles of lipids in brain functions, lipids acting as signaling molecules, and the advent of lipidomics enabled by the advances in mass spectrometry (MS) and liquid chromatography (LC). The identification and characterization of two classes of novel lipids from the brain are reviewed here: N-acyl phosphatidylserine (N-acyl-PS) and dolichoic acid (Dol-CA). The identification of these lipids benefited from the use of efficient lipid fractionation and separation techniques and highly sensitive, high-resolution tandem MS. PMID- 19303825 TI - Secretin-stimulated multi-detector CT versus mangafodipir trisodium-enhanced MR imaging plus MRCP in characterization of non-metastatic solid pancreatic lesions. AB - BACKGROUND AND AIM: Our study was aimed to compare multiphasic multi-detector computed tomography after secretin stimulation and mangafodipir trisodium enhanced magnetic resonance imaging plus MR cholangiopancreatography in the characterization of solid pancreatic lesions. PATIENTS AND METHODS: Forty patients with ultrasound diagnosis of solid pancreatic lesion prospectively underwent both multi-detector computed tomography and magnetic resonance imaging. Three minutes after intravenous administration of secretin, post-contrast computed tomography scans were performed 40, 80, and 180 s after contrast medium injection. MR protocol included axial/coronal, thin/thick-slab, single-shot T2 w sequences and axial/coronal T1 w breath-hold spoiled gradient-echo images before and 30-40 min after intravenous infusion of manganese dipyri-doxal diphosphate. Different observers blindly evaluated the ability of computed tomography and magnetic resonance imaging to characterize focal pancreatic lesions. Surgery, biopsy, and/or follow-up were considered as our diagnostic gold standard. RESULTS: Thirty-five focal pancreatic lesions (adenocarcinoma, n=18; focal chronic pancreatitis, n=4; endocrine tumor, n=6; metastasis, n=1; cystic tumor, n=3; indeterminate cystic lesions, n=3) were present in 34 patients since the remaining 6 subjects showed no pathological finding. Both multi-detector computed tomography and magnetic resonance imaging showed a statistically significant correlation with the gold standard and between themselves in the characterization of 29 solid lesions of the pancreas (p<0.05). CONCLUSION: Both imaging techniques well correlate to final diagnosis of non-metastatic solid pancreatic lesions and particularly of adenocarcinomas with a slight advantage for mangafodipir trisodium-enhanced magnetic resonance imaging plus MR cholangiopancreatography. PMID- 19303826 TI - [Association between late-onset sepsis and breast-milk colonization by Streptococcus agalactiae]. PMID- 19303827 TI - [Severe dyspnoea as a reason for a medical visit. Is it just a lung problem?]. PMID- 19303828 TI - [Adolescent drug use. Ethical dilemma in the diagnostic-therapeutic approach]. AB - Illegal drug use among adolescents has increased in recent years in Spain, as well as has the risk behaviours and problems typical of adolescence. The results of studies on drug use during this stage of life reveal the serious and wide ranging consequences that can arise. Emergency services are often the first to receive and deal with these and its professionals must face situations that pose contradictions between two of the basic ethical principles, the principle of autonomy and the principle of beneficence; an ethical dilemma that is addressed in this work. PMID- 19303829 TI - [Infant metabolic alkalosis of dietetic origin]. AB - Cases of metabolic alkalosis are divided into susceptible or resistant to treatment with sodium chloride, depending on the response to it. The resistant cases present with high urinary excretion of chloride, and are secondary to tubular disease or use of diuretics. Included among the sensitive cases are, vomiting, cystic fibrosis and low intake. Two infants were fed with "almond milk" and showed clinical symptoms of dehydration and failure to thrive. Hypochloraemic and hypokalaemic metabolic alkasosis was seen in both cases, which responded satisfactorily to water and electrolyte replacement. After ruling out vomiting, ingestion of drugs, tubular disease, and cystic fibrosis, the diagnosis was low intake, due to poor contribution of Na+ and Cl(-) provided by the "almond milk". This deficit induces an increase in proximal tubular reabsorption of H(-)CO3 and in parts of the distal nephron, an increase in reabsorption of Na+ and Cl(-) which are exchanged with K+ and H+, which can give rise to a hypochloraemic alkalosis and hypokalaemia. Secondary hypothyroidism was found in one case, apparently due to the lack of iodine in the almond milk, and disorders of the myelination characterized by optic neuritis and hearing loss. These disorders were resolved when feeding with human formula was established. PMID- 19303830 TI - Tuberculosis and tuberculosis/HIV co-infection in pregnancy. AB - Both tuberculosis (TB) and human immunodeficiency virus (HIV) affect women aged 15-29 years. This is the prime childbearing age group with an increasing mortality due to HIV. The key to the prevention of neonatal TB among these women is early recognition of TB, based primarily on maternal history and relevant investigations of the mother and newborn. There are World Health Organization (WHO) guidelines for maternal prophylaxis and therapy and prophylaxis to the newborn on the stage of the maternal disease. In HIV-infected women, CD4 counts should be monitored urgently as a guide to antiretroviral (ARV) prophylaxis. When the CD4 count is <200 cells/mm(3), WHO recommends that the mother should be treated with combination antiretroviral therapy (cART). In resource-rich settings most guidelines recommend treatment with cART when the CD4 count is <350 cells/mm(3). The combination of ARVs and anti-TB therapy poses difficulties which can be resolved by combination of different drugs. In both conditions, evidence suggests that in resource-limited settings exclusive breastfeeding is recommended, with the addition of flash heating of the mothers' milk for HIV infected women. PMID- 19303831 TI - Salvage reconstruction of the oesophagus: a retrospective study of 15 cases. AB - Salvage reconstruction of the oesophagus is still considered a challenging procedure for all head and neck surgeons. The risk of postoperative infection and delayed wound healing is high because of thick scar formation and persistent inflammation. Furthermore, recipient vessels for free tissue transfer or vascular supercharge are not always available. Alimentary tract reconstruction with skin or musculocutaneous flap may be necessary, but this method is susceptible to fistula formation.[Nakatsuka T, Harii K, Asato H, et al. Comparative evaluation in pharyngo-oesophageal reconstruction: radial forearm flap compared with jejunal flap. A 10-year experience. Scand J Plast Reconstr Surg Hand Surg 1998; 32: 307 10] In the past 10 years, we have experienced 15 cases of salvage reconstruction of the oesophagus after prior cancer treatment or aorto-oesophageal fistula; the cervical oesophagus was reconstructed in five cases and the cervico-thoracic oesophagus in 10. In four cases of cervical oesophagus and six of cervico thoracic oesophagus we performed free jejunal transfer including two long segment transfers with double vascular pedicle. The cervico-thoracic oesophagus was also reconstructed with pedicled alimentary tract transfer (colon interposition or jejunal pull-up) with vascular supercharge in four cases. In one case, cervical oesophageal defect was reconstructed with a latissimus dorsi musculocutaneous flap. We also used a deltopectoral flap to cover the skin defect in three cases. In three cases, a second salvage operation was necessary because of flap necrosis that was caused by unreliable recipient vessels resulting from scar formation and persistent inflammation. Successful restoration of the oesophagus and oral alimentation was achieved in 11 cases. From this study, we concluded that free jejunal transfer is a useful procedure for salvage reconstruction of the oesophagus, particularly for cervical oesophagus or short oesophageal defects. Nonetheless, surgeons should know the indications and limitations of this procedure thoroughly and always be ready to choose other reconstructive options if necessary. PMID- 19303832 TI - The pedicled descending branch muscle-sparing latissimus dorsi flap for trunk and upper extremity reconstruction. AB - BACKGROUND: The major blood supply of the latissimus dorsi muscle flap is based on the descending and tranverse branches of the thoracodorsal artery. This segmental blood supply allows the muscle to be split and harvested based solely on vascularization from the descending branch, thus sparing the latissimus dorsi muscle function. This article reports the use of the descending branch muscle sparing latissimus dorsi myocutaneous flap in reconstructing defects on the trunk and upper extremities. METHODS: Five patients with defects on the trunk or upper extremities had soft tissue reconstruction with a pedicled descending branch muscle-sparing latissimus dorsi myocutaneous flap. A transverse skin paddle design was used in all cases. All flaps were performed by the senior author. Complications were recorded, and range of motion analysis was performed comparing operated and non-operated sides during follow-up appointments. RESULTS: The descending branch muscle-sparing latissimus dorsi flap was used for reconstruction of: the chest wall (2), axilla (2) and upper extremity (1). The skin paddles harvested ranged from 15 x 7cm to 24 x 9cm. All donor sites were closed primarily. There was one case of minor wound dehiscence on the donor site and one case of wound infection (reconstruction was for chronic, severe axillary hidradenitis suppuritiva). There were no incidences of seroma. In all cases, there was no difference in strength or range of motion around the shoulder joint when comparing the operated to the non-operated side. CONCLUSION: The pedicled descending branch muscle-sparing latissimus dorsi myocutaneous flap with a transversely orientated skin paddle results in minimal functional deficit of the donor site, absence of seroma, low rate of flap complications and an aesthetically acceptable scar. PMID- 19303833 TI - Discussion of "Effects of hyperbaric oxygen on the replanted extremity subjected to prolonged warm ischemia". PMID- 19303834 TI - Current scales for assessing human scarring: a review. AB - Patients can have wide-ranging problems related to scars, in terms of cosmesis, function, symptoms, psychological problems and overall quality of life issues. A range of treatments have been recommended for problematic scarring, however it has been acknowledged that the evidence base for most of the recommendations for scar therapy is limited, with few studies using validated measures of scar assessment in generating data. This review critically evaluates the subjective scar assessment scales developed to date and provides an insight into developments required in this area for the future. The principles of psychometric theory are discussed as a means of developing reliable and valid outcome measures and these are also applicable for measuring outcomes in other fields of plastic surgery research. PMID- 19303835 TI - Use of Aquamid as a filler for facial rejuvenation in orientals. AB - We used Aquamid as a filler for facial augmentation and rejuvenation in Orientals. This article introduces the injection techniques, effects, adequate dosage and complications of this filler, especially about rejuvenation of nasolabial fold and nasojugal groove. From December 2002 to June 2007, 5676 patients were treated in our clinic group. Complications were relatively minimal (0.082%) in comparison to other fillers and long-term effects were revealed. This is the first report concerning Aquamid use in facial rejuvenation of the Orientals. PMID- 19303836 TI - Transsupraspinatus arthrotomy through an enlarged transacromial approach for total shoulder replacement. AB - The authors describe a step-by-step technique for anatomic total shoulder arthroplasty using transsupraspinatus arthrotomy via the enlarged transacromial approach. This technique seems ideal to ensure adequate postoperative tensional balance of the infraspinatus and the subscapularis, which is critical for the rotator cuffs to function properly and to achieve optimal arthroplasty stability. Reviewing these different steps helps understanding each rotator cuff individual component's contribution to achieve optimal arthroplasty stability. PMID- 19303837 TI - The anabolic action of intermittent PTH in combination with cathepsin K inhibitor or alendronate differs depending on the remodeling status in bone in ovariectomized mice. AB - We hypothesized that the anabolic action of parathyroid hormone (PTH) with the anti-catabolic agents cathepsin K inhibitor and alendronate differs depending on the remodeling status in the bone. C57/BL/6J mice, 8 weeks of age, were subjected to ovariectomized (OVX) or sham surgery. At 6 weeks after surgery, the mice were treated with cathepsin K inhibitor, alendronate, or a vehicle (daily, for 8 weeks), with or without PTH (1-34) (5 times/week, for the last 4 weeks). We assessed the bone chemical markers of the serum and urine, bone mineral density (BMD), histomorphomery in the primary and secondary spongiosa of the proximal tibia after fluorescence labeling, primary cell culture, and mRNA expressions in bone marrow cells. Cathepsin K inhibitor and alendronate significantly increased the BMD and the bone volume of the primary and secondary spongiosa, with a reduction of the urinary C-telopeptide of type I collagen that was increased by OVX, respectively. Cathepsin K inhibitor augmented the anabolic action of PTH on the BMD and bone volume at both the primary and secondary spongiosa, while alendronate had the same effect on the BMD and bone volume only at the primary spongiosa. Cathepsin K inhibitor did not decrease serum osteocalcin with or without PTH, while alendronate did decrease it. Cathepsin K inhibitor did not decrease the values of osteoclast number or bone formation rate with or without PTH, while alendronate decreased those values and increased osteoclast apoptosis. The combination of PTH and cathepsin K inhibitor increased alkaline phosphatase positive CFU-f formation and c-fos, osterix, and osteocalcin mRNA expressions of bone marrow cells as well as PTH alone, while the combination of PTH and alendronate decreased those values. This study demonstrated that alendronate enhances the anabolic action of PTH at the primary spongiosa, but blunts it in the remodeling trabecular bone, while cathepsin K inhibitor enhances the action at both sites in OVX mice. In conclusion, the anabolic action of intermittent PTH in combination with cathepsin K inhibitor or alendronate differs depending on the remodeling status of bone in OVX mice. PMID- 19303838 TI - Building public-private research partnerships in troubled times. AB - The Indian government is launching a series of new initiatives to boost public private research partnerships and to jumpstart science-driven economic growth. But will the current global economic downturn hamper these efforts? PMID- 19303839 TI - DISC1 partners with GSK3beta in neurogenesis. AB - The protein DISC1, encoded by a gene implicated in schizophrenia susceptibility, regulates the development of postmitotic neurons. Mao et al. (2009) now report that DISC1 also regulates the proliferation of embryonic and adult neural progenitor cells through the GSK3beta/beta-catenin pathway, providing new insights into how susceptibility genes may contribute to the etiology of psychiatric disorders. PMID- 19303840 TI - Polycomb repression under the skin. AB - Polycomb group proteins control a hierarchy of gene expression during the differentiation of stem cells. In this issue, Ezhkova et al. (2009) show that repression mediated by Polycomb proteins controls the timing of differentiation of precursor cells of the epidermal lineage. PMID- 19303841 TI - A MAPK scaffold lends a helping hand. AB - The scaffold proteins of signaling pathways are thought to act as passive tethering devices bringing together catalytic components of signaling cascades. Good et al. (2009) now reveal that in the budding yeast the scaffold protein Ste5 acts as an allosteric activator of the mitogen-activated protein kinase Fus3, rendering it competent to be a kinase substrate for signal transmission. PMID- 19303842 TI - A gate keeper for axonal transport. AB - The axon and dendritic arbor of neurons require different sets of membrane proteins to carry out their functions. In this issue, Song et al. (2009) describe how a cytoplasmic diffusion barrier in the axon initial segment of rat hippocampal neurons ensures that only axonal (and not dendritic) membrane proteins enter the axon. PMID- 19303843 TI - Extreme genome repair. AB - Slade et al. (2009) describe in this issue how the genome of the bacterium Deinococcus radiodurans gets reassembled after being shattered by high-dose radiation. In contrast to the extreme nature of the damage, the steps of repair appear surprisingly ordinary. So, why can't all organisms carry out extreme genome repair? PMID- 19303844 TI - Rethinking ALS: the FUS about TDP-43. AB - Mutations in TDP-43, a DNA/RNA-binding protein, cause an inherited form of the neurodegenerative disease amyotrophic lateral sclerosis (ALS). Two recent studies (Kwiatkowski et al., 2009; Vance et al., 2009) now report that mutations in FUS/TLS, another DNA/RNA-binding protein, also trigger premature degeneration of motor neurons. TDP-43 and FUS/TLS have striking structural and functional similarities, implicating alterations in RNA processing as a key event in ALS pathogenesis. PMID- 19303845 TI - Auxin: a trigger for change in plant development. AB - The dynamic, differential distribution of the hormone auxin within plant tissues controls an impressive variety of developmental processes, which tailor plant growth and morphology to environmental conditions. Various environmental and endogenous signals can be integrated into changes in auxin distribution through their effects on local auxin biosynthesis and intercellular auxin transport. Individual cells interpret auxin largely by a nuclear signaling pathway that involves the F box protein TIR1 acting as an auxin receptor. Auxin-dependent TIR1 activity leads to ubiquitination-based degradation of transcriptional repressors and complex transcriptional reprogramming. Thus, auxin appears to be a versatile trigger of preprogrammed developmental changes in plant cells. PMID- 19303846 TI - Disrupted in schizophrenia 1 regulates neuronal progenitor proliferation via modulation of GSK3beta/beta-catenin signaling. AB - The Disrupted in Schizophrenia 1 (DISC1) gene is disrupted by a balanced chromosomal translocation (1; 11) (q42; q14.3) in a Scottish family with a high incidence of major depression, schizophrenia, and bipolar disorder. Subsequent studies provided indications that DISC1 plays a role in brain development. Here, we demonstrate that suppression of DISC1 expression reduces neural progenitor proliferation, leading to premature cell cycle exit and differentiation. Several lines of evidence suggest that DISC1 mediates this function by regulating GSK3beta. First, DISC1 inhibits GSK3beta activity through direct physical interaction, which reduces beta-catenin phosphorylation and stabilizes beta catenin. Importantly, expression of stabilized beta-catenin overrides the impairment of progenitor proliferation caused by DISC1 loss of function. Furthermore, GSK3 inhibitors normalize progenitor proliferation and behavioral defects caused by DISC1 loss of function. Together, these results implicate DISC1 in GSK3beta/beta-catenin signaling pathways and provide a framework for understanding how alterations in this pathway may contribute to the etiology of psychiatric disorders. PMID- 19303847 TI - The BRC repeats of BRCA2 modulate the DNA-binding selectivity of RAD51. AB - The breast cancer susceptibility protein, BRCA2, is essential for recombinational DNA repair. BRCA2 delivers RAD51 to double-stranded DNA (dsDNA) breaks through interaction with eight conserved, approximately 35 amino acid motifs, the BRC repeats. Here we show that the solitary BRC4 promotes assembly of RAD51 onto single-stranded DNA (ssDNA), but not dsDNA, to stimulate DNA strand exchange. BRC4 acts by blocking ATP hydrolysis and thereby maintaining the active ATP-bound form of the RAD51-ssDNA filament. Single-molecule visualization shows that BRC4 does not disassemble RAD51-dsDNA filaments but rather blocks nucleation of RAD51 onto dsDNA. Furthermore, this behavior is manifested by a domain of BRCA2 comprising all eight BRC repeats. These results establish that the BRC repeats modulate RAD51-DNA interaction in two opposing but functionally reinforcing ways: targeting active RAD51 to ssDNA and prohibiting RAD51 nucleation onto dsDNA. Thus, BRCA2 recruits RAD51 to DNA breaks and, we propose, the BRC repeats regulate DNA-binding selectivity. PMID- 19303848 TI - Recombination and replication in DNA repair of heavily irradiated Deinococcus radiodurans. AB - Deinococcus radiodurans' extreme resistance to ionizing radiation, desiccation, and DNA-damaging chemicals involves a robust DNA repair that reassembles its shattered genome. The repair process requires diploidy and commences with an extensive exonucleolytic erosion of DNA fragments. Liberated single-stranded overhangs prime strand elongation on overlapping fragments and the elongated complementary strands reestablish chromosomal contiguity by annealing. We explored the interdependence of the DNA recombination and replication processes in the reconstitution of the D. radiodurans genome disintegrated by ionizing radiation. The priming of extensive DNA repair synthesis involves RecA and RadA proteins. DNA polymerase III is essential for the initiation of repair synthesis, whereas efficient elongation requires DNA polymerases I and III. Inactivation of both polymerases leads to degradation of DNA fragments and rapid cell death. The present in vivo characterization of key recombination and replication processes dissects the mechanism of DNA repair in heavily irradiated D. radiodurans. PMID- 19303849 TI - A role of DNA-PK for the metabolic gene regulation in response to insulin. AB - Fatty acid synthase (FAS) is a central enzyme in lipogenesis and transcriptionally activated in response to feeding and insulin signaling. The transcription factor USF is required for the activation of FAS transcription, and we show here that USF phosphorylation by DNA-PK, which is dephosphorylated by PP1 in response to feeding, triggers a switch-like mechanism. Under fasting conditions, USF-1 is deacetylated by HDAC9, causing promoter inactivation. In contrast, feeding induces the recruitment of DNA-PK to USF-1 and its phosphorylation, which then allows recruitment of P/CAF, resulting in USF-1 acetylation and FAS promoter activation. DNA break/repair components associated with USF induce transient DNA breaks during FAS activation. In DNA-PK-deficient SCID mice, feeding-induced USF-1 phosphorylation/acetylation, DNA breaks, and FAS activation leading to lipogenesis are impaired, resulting in decreased triglyceride levels. Our study demonstrates that a kinase central to the DNA damage response mediates metabolic gene activation. PMID- 19303850 TI - Protein acetylation microarray reveals that NuA4 controls key metabolic target regulating gluconeogenesis. AB - Histone acetyltransferases (HATs) and histone deacetylases (HDACs) conduct many critical functions through nonhistone substrates in metazoans, but only chromatin associated nonhistone substrates are known in Saccharomyces cerevisiae. Using yeast proteome microarrays, we identified and validated many nonchromatin substrates of the essential nucleosome acetyltransferase of H4 (NuA4) complex. Among these, acetylation sites (Lys19 and 514) of phosphoenolpyruvate carboxykinase (Pck1p) were determined by tandem mass spectrometry. Acetylation at Lys514 was crucial for enzymatic activity and the ability of yeast cells to grow on nonfermentable carbon sources. Furthermore, Sir2p deacetylated Pck1p both in vitro and in vivo. Loss of Pck1p activity blocked the extension of yeast chronological life span caused by water starvation. In human hepatocellular carcinoma (HepG2) cells, human Pck1 acetylation and glucose production were dependent on TIP60, the human homolog of ESA1. Our findings demonstrate a regulatory function for the NuA4 complex in glucose metabolism and life span by acetylating a critical metabolic enzyme. PMID- 19303851 TI - The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. AB - The scaffold protein Ste5 is required to properly direct signaling through the yeast mating pathway to the mitogen-activated protein kinase (MAPK), Fus3. Scaffolds are thought to function by tethering kinase and substrate in proximity. We find, however, that the previously identified Fus3-binding site on Ste5 is not required for signaling, suggesting an alternative mechanism controls Fus3's activation by the MAPKK Ste7. Reconstituting MAPK signaling in vitro, we find that Fus3 is an intrinsically poor substrate for Ste7, although the related filamentation MAPK, Kss1, is an excellent substrate. We identify and structurally characterize a domain in Ste5 that catalytically unlocks Fus3 for phosphorylation by Ste7. This domain selectively increases the k(cat) of Ste7-->Fus3 phosphorylation but has no effect on Ste7-->Kss1 phosphorylation. The dual requirement for both Ste7 and this Ste5 domain in Fus3 activation explains why Fus3 is selectively activated by the mating pathway and not by other pathways that also utilize Ste7. PMID- 19303852 TI - Specific recognition of linear ubiquitin chains by NEMO is important for NF kappaB activation. AB - Activation of nuclear factor-kappaB (NF-kappaB), a key mediator of inducible transcription in immunity, requires binding of NF-kappaB essential modulator (NEMO) to ubiquitinated substrates. Here, we report that the UBAN (ubiquitin binding in ABIN and NEMO) motif of NEMO selectively binds linear (head-to-tail) ubiquitin chains. Crystal structures of the UBAN motif revealed a parallel coiled coil dimer that formed a heterotetrameric complex with two linear diubiquitin molecules. The UBAN dimer contacted all four ubiquitin moieties, and the integrity of each binding site was required for efficient NF-kappaB activation. Binding occurred via a surface on the proximal ubiquitin moiety and the canonical Ile44 surface on the distal one, thereby providing specificity for linear chain recognition. Residues of NEMO involved in binding linear ubiquitin chains are required for NF-kappaB activation by TNF-alpha and other agonists, providing an explanation for the detrimental effect of NEMO mutations in patients suffering from X-linked ectodermal dysplasia and immunodeficiency. PMID- 19303853 TI - A phosphoinositide switch controls the maturation and signaling properties of APPL endosomes. AB - The recent identification of several novel endocytic compartments has challenged our current understanding of the topological and functional organization of the endocytic pathway. Using quantitative single vesicle imaging and acute manipulation of phosphoinositides we show that APPL endosomes, which participate in growth factor receptor trafficking and signaling, represent an early endocytic intermediate common to a subset of clathrin derived endocytic vesicles and macropinosomes. Most APPL endosomes are precursors of classical PI3P positive endosomes, and PI3P plays a critical role in promoting this conversion. Depletion of PI3P causes a striking reversion of Rab5 positive endosomes to the APPL stage, and results in enhanced growth factor signaling. These findings reveal a surprising plasticity of the early endocytic pathway. Importantly, PI3P functions as a switch to dynamically regulate maturation and signaling of APPL endosomes. PMID- 19303854 TI - Ezh2 orchestrates gene expression for the stepwise differentiation of tissue specific stem cells. AB - Although in vitro studies of embryonic stem cells have identified polycomb repressor complexes (PRCs) as key regulators of differentiation, it remains unclear as to how PRC-mediated mechanisms control fates of multipotent progenitors in developing tissues. Here, we show that an essential PRC component, Ezh2, is expressed in epidermal progenitors but diminishes concomitant with embryonic differentiation and with postnatal decline in proliferative activity. We show that Ezh2 controls proliferative potential of basal progenitors by repressing the Ink4A-Ink4B locus and tempers the developmental rate of differentiation by preventing premature recruitment of AP1 transcriptional activator to the structural genes that are required for epidermal differentiation. Together, our studies reveal that PRCs control epigenetic modifications temporally and spatially in tissue-restricted stem cells. They maintain their proliferative potential and globally repressing undesirable differentiation programs while selectively establishing a specific terminal differentiation program in a stepwise fashion. PMID- 19303855 TI - Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration. AB - Interactions between developmental signaling pathways govern the formation and function of stem cells. Prostaglandin (PG) E2 regulates vertebrate hematopoietic stem cells (HSC). Similarly, the Wnt signaling pathway controls HSC self-renewal and bone marrow repopulation. Here, we show that wnt reporter activity in zebrafish HSCs is responsive to PGE2 modulation, demonstrating a direct interaction in vivo. Inhibition of PGE2 synthesis blocked wnt-induced alterations in HSC formation. PGE2 modified the wnt signaling cascade at the level of beta catenin degradation through cAMP/PKA-mediated stabilizing phosphorylation events. The PGE2/Wnt interaction regulated murine stem and progenitor populations in vitro in hematopoietic ES cell assays and in vivo following transplantation. The relationship between PGE2 and Wnt was also conserved during regeneration of other organ systems. Our work provides in vivo evidence that Wnt activation in stem cells requires PGE2, and suggests the PGE2/Wnt interaction is a master regulator of vertebrate regeneration and recovery. PMID- 19303856 TI - Coincident pre- and postsynaptic activation induces dendritic filopodia via neurotrypsin-dependent agrin cleavage. AB - The synaptic serine protease neurotrypsin is essential for cognitive function, as its deficiency in humans results in severe mental retardation. Recently, we demonstrated the activity-dependent release of neurotrypsin from presynaptic terminals and proteolytical cleavage of agrin at the synapse. Here we show that the activity-dependent formation of dendritic filopodia is abolished in hippocampal neurons from neurotrypsin-deficient mice. Administration of the neurotrypsin-dependent 22 kDa fragment of agrin rescues the filopodial response. Detailed analyses indicated that presynaptic action potential firing is necessary for the release of neurotrypsin, whereas postsynaptic NMDA receptor activation is necessary for the neurotrypsin-dependent cleavage of agrin. This contingency characterizes the neurotrypsin-agrin system as a coincidence detector of pre- and postsynaptic activation. As the resulting dendritic filopodia are thought to represent precursors of synapses, the neurotrypsin-dependent cleavage of agrin at the synapse may be instrumental for a Hebbian organization and remodeling of synaptic circuits in the CNS. PMID- 19303857 TI - SnapShot: Auxin signaling and transport. PMID- 19303858 TI - Ultrasensitive electrochemical immunosensor for ochratoxin A using gold colloid mediated hapten immobilization. AB - A convenient, specific, and highly sensitive electrochemical immunosensor based on an indirect competitive assay format was developed for the determination of ochratoxin A (OTA), a common toxic contaminant in various kinds of agricultural products. The sensing substrate was prepared using a gold electrode modified with a self-assembled monolayer of 1,6-hexanedithiol that mediated the assembly of a gold colloid layer, which could enhance the surface loading of OTA-ovalbumin conjugate and improve the sensitivity in electrochemical readouts. After competition of the limited anti-OTA mouse monoclonal antibody between immobilized hapten and OTA analyte in sample solution, alkaline phosphatase (ALP)-labeled horse anti-mouse immunoglobulin G (IgG) antibody was selectively bound onto the surface of the electrode, affording an indicator for OTA concentration in the sample. Electrochemical response arising from the oxidation of enzymatic product of 1-naphthyl phosphate was observed to be inversely proportional to OTA concentration in the range from 10 pg/ml to 100 ng/ml with a detection limit as low as 8.2 pg/ml. Furthermore, a negligible matrix effect and good recoveries were obtained in the determination of corn samples, evidencing the feasibility of the proposed method for accurate determination of OTA in corn samples. PMID- 19303859 TI - Characterization of a fluorophore binding RNA aptamer by fluorescence correlation spectroscopy and small angle X-ray scattering. AB - Using fluorescence correlation spectroscopy (FCS), we have established an in vitro assay to study RNA dynamics by analyzing fluorophore binding RNA aptamers at the single molecule level. The RNA aptamer SRB2m, a minimized variant of the initially selected aptamer SRB-2, has a high affinity to the disulfonated triphenylmethane dye sulforhodamine B. A mobility shift of sulforhodamine B after binding to SRB2m was measured. In contrast, patent blue V (PBV) is visible only if complexed with SRB2m due to increased molecular brightness and minimal background. With small angle X-ray scattering (SAXS), the three-dimensional structure of the RNA aptamer was characterized at low resolution to analyze the effect of fluorophore binding. The aptamer and sulforhodamine B-aptamer complex was found to be predominantly dimeric in solution. Interaction of PBV with SRB2m led to a dissociation of SRB2m dimers into monomers. Radii of gyration and hydrodynamic radii, gained from dynamic light scattering, FCS, and fluorescence cross-correlation experiments, led to comparable conclusions. Our study demonstrates how RNA-aptamer fluorophore complexes can be simultaneously structurally and photophysically characterized by FCS. Furthermore, fluorophore binding RNA aptamers provide a tool for visualizing single RNA molecules. PMID- 19303860 TI - Determination of CYP2D6 gene copy number by multiplex polymerase chain reaction analysis. AB - Cytochrome P450 2D6 (CYP2D6) copy number variation (CNV) influences the metabolism of 15-25% of clinical drugs. Here we describe a novel multiplex polymerase chain reaction (PCR) analysis method that accurately detects CYP2D6 CNV and CYP2D6*9 allele. It includes the amplification of 2 CYP2D6 and 7 control (AQP1, CYP3A4, MDR1, and SDHB) fluorescent PCR products that are separated on a capillary sequencer and normalized using reference samples. The technique was validated using 27 PCR-restriction fragment length polymorphism (RFLP) pregenotyped samples and further tested in 75 Caucasian samples. The method assigns the correct CYP2D6 copy number, independent of already characterized CYP2D6 single nucleotide polymorphisms (SNPs), and could easily be applied to clinical samples. PMID- 19303861 TI - Gold immunochromatographic assay for simultaneous detection of carbofuran and triazophos in water samples. AB - Using a simple test for rapid identification and quantification of pesticide multiresidues in food and environmental samples is a long-cherished approach for practical monitoring purposes. Here two gold-based lateral-flow strips (strip A and strip B) were investigated for simultaneous detection of carbofuran and triazophos. For the strip A format, a bispecific monoclonal antibody (BsMcAb) against both carbofuran and triazophos was employed to prepare the immunogold probe. For the strip B format, anti-carbofuran monoclonal antibody (McAb) and anti-triazophos McAb separately labeled with colloidal gold were combined as detector reagents. By comparison of visual results from pesticide standard tests between the two formats, the strip B assay manifested higher sensitivities for both pesticides. Analysis of spiked water samples by the preferable strip indicated that the detection limits for carbofuran and triazophos were 32 and 4 microg/L, respectively. The strength of the portable one-step strip assay was in the simultaneous screening for two pesticides within a short time (8-10 min) without any equipment. PMID- 19303862 TI - Transcriptional regulation of the human Na+/H+ exchanger NHE3 by serotonin in intestinal epithelial cells. AB - Serotonin (5-HT) decreases NHE2 and NHE3 activities under acute conditions in human intestinal epithelial cells. Here, we have investigated the effects of 5-HT on expression of the human NHE3 gene and the mechanisms underlying its transcriptional regulation in differentiated C2BBe1 cells. Treatment of the human intestinal epithelial cell line, C2BBe1, with 5-HT (20 microM) resulted in a significant decrease in NHE3 mRNA and protein expression. In transient transfection studies, 5-HT repressed the NHE3 promoter activity by approximately 55%. The repression of the NHE3 promoter activity in response to 5-HT was accompanied by reduced DNA-binding activity of transcription factors Sp1 and Sp3 to the NHE3 promoter without alteration in their nuclear levels. Pharmacological inhibitors of protein kinase C reversed the inhibitory effect of 5-HT on the promoter activity. Our data indicate that 5-HT suppresses the transcriptional activity of the NHE3 promoter and this effect may be mediated by PKCalpha and modulation of DNA-binding affinities of Sp1 and Sp3. PMID- 19303863 TI - PI3 kinase/Akt signaling mediates epithelial-mesenchymal transition in hypoxic hepatocellular carcinoma cells. AB - Hypoxia activates genetic programs that facilitate cell survival; however, in cancer, it may promote invasion and metastasis. Although the exact mechanisms driving hypoxia-induced invasion and metastasis remain elusive, we hypothesized that epithelial-mesenchymal transition (EMT) may play a major role. We investigated this in vitro by treating hepatocellular carcinoma cells under 1.0% O(2). After the hypoxia treatment, the cells exhibited some morphological changes including cell elongation, cytoskeletal rearrangement, and junctional disruption. Moreover, expression of the epithelia-specific marker E-cadherin was decreased and expression of the myofibroblast-specific marker vimentin was detected in the treated cells. Cell migration and ECM gel invasion were increased. These findings were consistent with events observed during EMT. Hypoxia-induced EMT is accompanied by increased phosphorylation, activation of Akt and the downstream signaling. Hypoxia-induced EMT was blocked by PI3K inhibitor LY294002. The results suggest that the PI3K/Akt-dependent signaling pathways serve to regulate hypoxia-induced EMT of hepatocellular carcinoma cells. PMID- 19303864 TI - Retinal light input is required to sustain plasma melatonin rhythms in Nile tilapia Oreochromis niloticus niloticus. AB - The aim of this work was to confirm previous findings suggesting that the eyes are required for night-time melatonin production in Nile tilapia and further characterise this divergent circadian organisation. To do so, melatonin levels were firstly measured in eyecups and plasma to determine circadian patterns of melatonin production. Secondly, the effect of partial ophthalmectomy on the suppression of melatonin production was determined in vivo as well as ex vivo pineal light/dark sensitivity. Finally, to investigate whether such findings could be related to post-surgery stress, melatonin analyses were performed in the subsequent 24 h and 7 days post-ophthalmectomy with cortisol levels assessed as an indicator of stress. Our results showed an inverse pattern of melatonin production in the eye cups of tilapia compared to blood circulating levels, suggesting different roles played by melatonin in these two tissues. Results then demonstrated that total or partial ophthalmectomy resulted in the suppression of night-time melatonin production. Furthermore, although pineals in culture were shown to be photosensitive, night-time melatonin levels were much lower than seen in other species. Finally, when performing sampling immediately or one week post surgery, no difference in the melatonin profiles were observed. It is therefore unlikely that post-surgery stress would explain such suppression in melatonin production although all fish displayed high cortisol levels most probably due to social and handling stress. Taken together, these results provide further evidence of a new type of circadian organisation in a teleost species where the eyes are required to sustain night-time melatonin levels. PMID- 19303865 TI - Electrophysiological assessment of the brain function in term SGA infants. AB - Small for gestational age (SGA) infants are defined as babies having a birth weight below the 10th percentile for gestational age. A great number of studies have shown that children with SGA have an increased risk of impaired neurodevelopment. Electroencephalography (EEG) is an excellent method for measuring brain maturation in newborns. In this study, the effect of SGA on the maturation of cerebrocortical electrographic activity was investigated by the EEG and also analyzed with power spectral analysis. Serial EEGs were performed in 40 term SGAs, and 20 term appropriate for gestational age (AGA) infants in 1st week, 1st and 3rd month. Power spectral analysis was performed quantitatively in five channels (Fp1-C3, C3-O1, Fp2-C4, C4-O2, and Cz-C4 channels). Amplitude levels of the SGA group were significantly lower than the AGA group in all records. Delta frequency was the major frequency component in the groups. Delta frequency activities in the midline vertex region were decreased in the AGA group with increasing postconceptual age while the activities of the SGA group were increased. Contrarily, beta frequency activities in the midline vertex region were increased in the AGA group with increasing postconceptual age while these activities of the SGA group were decreased. Theta frequency activities in the fronto-central regions were lower in the SGA group. In terms of the vertex, k complex, and sleep spindle, there was no difference between the two groups. We conclude that cerebrocortical electrophysiological maturation has been delayed in term SGA infants during the first three months of postnatal life. PMID- 19303866 TI - A solid-phase method for preparing human DNA from urine for diagnostic purposes. AB - OBJECTIVES: To develop a simple method using paramagnetic beads for isolation of human DNA from small volumes of urine. The method should be amendable for automation. The purified DNA is intended to be used in downstream diagnostics and screening studies using nucleic acid amplification techniques. DESIGN AND METHODS: Unspecific capture of cells present in urine to magnetic particles, lysis and subsequent binding of the DNA to the same bead surface. RESULTS: DNA isolated using the method could be used as template for sensitive real-time PCR and end-point PCR using primers targeted to the GAPDH, K-ras, DD3 and p53 genes. Compared to silica spin column-based extraction, the method showed equal or higher DNA yields. The method performed reliably when automated using a liquid handling robot equipped with a magnetic workstation. CONCLUSIONS: The method generates purified DNA free from inhibitors, applicable for sensitive applications such as real-time PCR, genotyping, and for sequence variant analysis. The use of magnetic beads allows for automation, reducing hands-on time and creating a high throughput and reproducible protocol for the purpose of large scale screening and diagnostics. PMID- 19303867 TI - The effects of sildenafil on the functional and structural changes of ileum induced by intestinal ischemia-reperfusion in rats. AB - There is evidence demonstrating the protective effect of cGMP-specific phosphodiesterase type 5 (PDE5) inhibitors against ischemic injury in certain tissues. In this study, sildenafil, a potent inhibitor of PDE5, was tested for its beneficial effects in the prevention of disrupted ileal contractility and damage to tissue caused by intestinal ischemia-reperfusion in rats. Male Sprague Dawley rats were divided into four groups: sham-operated; sham-operated with sildenafil pretreatment; ischemia-reperfusion with vehicle pretreatment; and ischemia-reperfusion with sildenafil pretreatment. The superior mesenteric artery was occluded for 45 min to induce ischemia. The clamp was then removed for a 60 min period of reperfusion. Sildenafil (1 mg/kg, i.v.) or saline was administered prior to the surgical procedure in the ischemia-reperfusion and sham-operated groups. Isometric contractions of the ileal segments in response to acetylcholine or electrical field stimulation (120 V, 2 ms pulse for 5 s, 1-20 Hz) were recorded. Additionally, levels of thiobarbituric acid reactive substances and myeloperoxidase activity were measured in addition to a histopathological examination of the ileal tissue. The contractions induced by both acetylcholine and electrical field stimulations were markedly inhibited after ischemia reperfusion. Sildenafil pretreatment (1 mg/kg, i.v.) abolished the inhibition of responses to acetylcholine. The increased levels of thiobarbituric acid reactive substances and myeloperoxidase activity caused by ischemia-reperfusion were reversed to control levels with sildenafil pretreatment. Intestinal ischemia reperfusion caused severe ischemic injury in rat ileum, which was prevented by sildenafil. These results suggest that sildenafil pretreatment has a protective effect against ileal dysfunction and damage induced by intestinal ischemia reperfusion in the rat. PMID- 19303868 TI - Structure-function relationship studies of PTH(1-11) analogues containing D-amino acids. AB - Parathyroid hormone (PTH) is an 84-amino acid peptide hormone. Produced in the parathyroid glands, it acts primarily on bone and kidney to maintain extracellular calcium levels within normal limits. It has been shown that the 1 34 amino acid fragment of PTH is sufficient to bind and activate the PTH type-I receptor. Recent investigations focusing on the interaction of N-terminal fragments of PTH with PTH type-I receptor showed that certain modifications can increase signalling potency in peptides as short as 11 amino acids. To understand the role of the side chains of all the amino acid residues in PTH(1-11), we synthesized all-D PTH, three retro-inverso analogues of the most active modified PTH(1-11), H-Aib-Val-Aib-Glu-Ile-Gln-Leu-Nle-His-Gln-Har-NH(2), and we substituted every L-AA of the latter with the corresponding D-AA, obtaining a library of PTH(1-11) analogues that were tested as agonists. The library was synthesized by SPPS, employing the Fmoc protocol. The biological tests showed that the activity of the D-Har11 analogue is of the same order of magnitude of that of the most active modified PTH(1-11). This behaviour is paralleled by an increase of the helical content on going from the D-Val(2) to the D-Har(11) analogue. This is in agreement with previous work where a correlation between activity and helical content has been demonstrated. The importance of a positively charged group in the C-terminal position is shown to be independent of the configuration of the C(alpha)-carbon. PMID- 19303869 TI - Honokiol protects rats against eccentric exercise-induced skeletal muscle damage by inhibiting NF-kappaB induced oxidative stress and inflammation. AB - Honokiol, a bioactive component isolated from the Chinese herb Magnolia officinalis, is known for its potent antioxidative and anti-inflammatory effects. To study whether honokiol can protect skeletal muscle from sports injuries, we set up an eccentric exercise bout protocol for rats consisting of downhill running on a treadmill and examined the effect of oral administration of honokiol at 1 h before eccentric exercise at a dose of 5 mg/kg on day 1 (HK5 x 1) or 1 mg/kg/day for 5 consecutive days (HK1 x 5). Eccentric exercise was implemented for 3-5 consecutive days, and induced remarkable tissue damage. This damage was associated with an increase in serum creatine levels, increase in protein nitrotyrosylation, poly-ADP-ribose-polymerase (PARP) upregulation, lipid peroxidation, and leukocyte infiltration. The degree of muscle damage also paralleled dramatic gene expression for cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and inflammation-associated cytokines (interleukin (IL)-1beta, IL-6, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1), possibly through activation of nuclear factor kappa-B (NF-kappaB), a crucial proinflammatory transcription factor. Both honokiol treatments (HK5 x 1 and HK1 x 5) significantly ameliorated eccentric exercise-induced muscle damage as revealed by suppression of cell fragmentation, protein nitrotyrosylation and PARP upregulation, as well as reductions in lipid peroxidation and leukocyte infiltration, possibly through downregulating gene expression for COX-2, iNOS, and the proinflammatory cytokines by modulation of NF-kappaB activation. In conclusion, the present study demonstrates for the first time that honokiol exhibits protective effects against eccentric exercise-induced skeletal muscle damage in rats, probably by modulating inflammation-mediated damage to muscle cells. PMID- 19303870 TI - Icariin enhances neuronal survival after oxygen and glucose deprivation by increasing SIRT1. AB - It has been reported that icariin protects neurons against ischemia/reperfusion injury. In this study, we found that icariin could enhance neuronal viability and suppress neuronal death after oxygen and glucose deprivation (OGD). Further study showed that neuroprotection by icariin was through the induction of Sirtuin type 1 (SIRT1), an effect that was reversed by SIRT1 inhibitor III and P38 inhibitor SB203580. SIRT1 is an endogenous gene of longevity, which increased neuronal viability and could be activated by stimulating the mitogen-activated protein kinase (MAPK) pathway. However, this study found that icariin activated the MAPK/P38 pathway, not the extracellular signal-regulated kinase (MAPK/ERK) or c Jun N-terminal protein kinase (MAPK/JNK) to regulate SIRT1 expression. The results suggest that icariin may be developed into a neuroprotectant for ischemia related brain injury. PMID- 19303871 TI - Leishmania (Viannia) panamensis: an in vitro assay using the expression of GFP for screening of antileishmanial drug. AB - Promastigotes of Leishmania (Viannia) panamensis were successfully transfected with p6.5-egfp to express green fluorescent protein. The transfectants remained infective to macrophages, providing an in vitro model for screening antileishmanial drugs. This was demonstrated by flow cytometry of macrophage associated GFP after exposure of infected cultures to known antileishmanial drugs, i.e. amphotericin B and glucantime. Fluorescence of GFP diminished progressively from infected cells with increasing drug concentrations used in both cases. The availability of this fluorescent assay for infection of macrophages by L. (V.) panamensis facilitates drug discovery program for the Viannia species, which differ significantly from those of the Leishmania subgenus. PMID- 19303872 TI - Fasciola gigantica: immunolocalization of 28.5 kDa antigen in the tegument of metacercaria and juvenile fluke. AB - Specific monoclonal antibody (MoAb) to 28.5 kDa tegumental antigen (TA) was used to localize this antigen in the tissues of metacercariae, newly excysted juvenile (NEJ), 1, 3, 5, and 7-week-old juveniles of Fasciola gigantica by using indirect immunofluorescence, immunoperoxidase and immunogold techniques. Both indirect immunofluorescence and immunoperoxidase detections showed that this antigen was concentrated in the tegument particularly in its outer rim, tegumental cells and their processes as well as epithelial linings of the oral sucker. Unlike adult F. gigantica, it was not detected in spermatogenic cells in the testes, cells of Mehlis'gland, oocytes within the ovary, and ovum within the egg of parasites. At the ultrastructural level, the immunogold labeling showed deposit of gold particles specifically in G2 tegumental granules and on the surface membrane. Thus, this 28.5 kDa antigen is expressed in the tegument and associated structures of juvenile parasites, and it could be a major component of the G2 granules which are shown to fuse with the surface membrane and contribute material to replace the casted-off membrane. This process is the replenishment and turnover of the surface membrane to prevent the attachment of the host immune effector cells. PMID- 19303873 TI - Cardiac stem/progenitor cells, secreted proteins, and proteomics. AB - Stem cell-based therapy is emerging as a novel approach for myocardial repair over conventional cardiovascular therapies. In addition to embryonic stem cells and adult stem cells from noncardiac sources, there is a small population of resident stem cells in the heart from which new cardiac cells (myocytes, vascular endothelial cells and smooth muscle cells) can be derived and used for cardiac repair in case of heart injury. It has been proposed that the clinical benefit of stem cells may arise from secreted proteins that mediate regeneration in a paracrine/autocrine manner. To be able to track the regulatory pathway on a molecular basis, utilization of proteomics in stem cell research is essential. Proteomics offers a tool that can address questions regarding stem cell response to disease/injury. PMID- 19303874 TI - Induction of gene encoding FABP4 in Pten-null keratinocytes. AB - Keratinocyte-specific Pten-null mice revealed distinct phenotypes, including epidermal and sebaceous gland hyperplasia. To determine the candidate genes that contribute to their phenotypes, we analyzed a comprehensive gene expression of Pten-null keratinocytes using microarray technology. Consequently, it was demonstrated that the most induced gene was adipocyte-specific fatty acid-binding protein (FABP4). Collectively, it is conceivable that the FABP4 pathway mediates the sebaceous gland hyperplasia in keratinocyte-specific Pten-null mice. PMID- 19303875 TI - Venoms, venomics, antivenomics. AB - Venoms comprise mixtures of peptides and proteins tailored by Natural Selection to act on vital systems of the prey or victim. Here we review our proteomic protocols for uncoiling the composition, immunological profile, and evolution of snake venoms. Our long-term goal is to gain a deep insight of all viperid venom proteomes. Knowledge of the inter- and intraspecies ontogenetic, individual, and geographic venom variability has applied importance for the design of immunization protocols aimed at producing more effective polyspecific antivenoms. A practical consequence of assessing the cross-reactivity of heterologous antivenoms is the possibility of circumventing the restricted availability of species-specific antivenoms in some regions. Further, the high degree of target specificity makes toxins valuable scaffolds for drug development. PMID- 19303876 TI - The Hypocrea jecorina (syn. Trichoderma reesei) lxr1 gene encodes a D-mannitol dehydrogenase and is not involved in L-arabinose catabolism. AB - The Hypocrea jecorina LXR1 was described as the first fungal L-xylulose reductase responsible for NADPH dependent reduction of L-xylulose to xylitol in L-arabinose catabolism. Phylogenetic analysis now reveals that LXR1 forms a clade with fungal D-mannitol 2-dehydrogenases. Lxr1 and the orthologous Aspergillus niger mtdA are not induced by L-arabinose but expressed at low levels during growth on different carbon sources. Deletion of lxr1 does not affect growth on L-arabinose and L xylulose reductase activity remains unaltered whereas D-mannitol 2-dehydrogenase activities are reduced. We conclude that LXR1 is a D-mannitol 2-dehydrogenase and that a true LXR1 is still awaiting discovery. PMID- 19303877 TI - Functional genomics and proteomics approaches to study the ERBB network in cancer. AB - Substantial progress in functional genomic and proteomic technologies has opened new perspectives in biomedical research. The sequence of the human genome has been mostly determined and opened new visions on its complexity and regulation. New technologies, like RNAi and protein arrays, allow gathering knowledge beyond single gene analysis. Increasingly, biological processes are studied with systems biological approaches, where qualitative and quantitative data of the components are utilized to model the respective processes, to predict effects of perturbations, and to then refine these models after experimental testing. Here, we describe the potential of applying functional genomics and proteomics, taking the ERBB family of growth-factor receptors as an example to study the signaling network and its impact on cancer. PMID- 19303878 TI - Hypoxia-inducible factor 1 alpha activates and is inhibited by unoccupied estrogen receptor beta. AB - Previously, we showed that hypoxia induces ligand-independent estrogen receptor (ER)alpha activation. In this study, we found that hypoxia activated the ER beta mediated transcriptional response in HEK293 cells in the absence of estrogen. ER beta transactivation was induced by the expression of the hypoxia-inducible factor 1 alpha (HIF-1 alpha) under normoxia. ER beta interacted with HIF-1 alpha, and SRC1 and CBP potentiated the effect of HIF-1 alpha on ER beta-mediated transcription. We then examined the effect of ER beta on HIF1-alpha transactivation. Surprisingly, ER beta attenuated the transcriptional activity of HIF-1 alpha, as measured by HRE-driven reporter gene expression and hypoxic induction of VEGF mRNA in HEK293 cells. Taken together, these data show that HIF 1 alpha activates ER beta-mediated transcription in the absence of a ligand, and ER beta inhibits HIF-1 alpha-mediated transcription. PMID- 19303879 TI - New hypotheses on the function of the avian shell gland derived from microarray analysis comparing tissue from juvenile and sexually mature hens. AB - Activation of the shell gland region of the avian oviduct is mediated by ovarian steroids. To understand more extensively how shell glands are maintained and function, we have compared gene expression in the shell glands from juvenile and laying hens using a chicken cDNA microarray. Average expression profiles of juvenile and sexually mature shell glands were compared resulting in the identification of 266 differentially regulated genes. Reverse transcription quantitative polymerase chain reaction confirmed expression differences. The differentially expressed genes included several with known involvement in shell gland function, including ion transport and shell matrix proteins. There were also many unpredicted differentially expressed genes, and for some we propose hypotheses for their functions. These include those encoding (a) osteoprotegerin, a decoy death receptor for receptor activator of nuclear factor NFkB ligand (RANKL) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), that in the shell gland, may prevent apoptosis and/or may have an endocrine effect by preventing RANKL's action on bone osteoclasts that mobilize stored calcium; (b) prostatic acid phosphatase (ACPP) and prostate stem cell antigen (PSCA) that could play a role in sperm physiology within the shell gland; (c) urea transporter (SLC14A2) that could provide a novel anti-microbial defence; (d) bactericidal/permeability-increasing protein-like 2 (BPIL2), and other potential anti-microbials that have not previously been documented in the chicken. These new hypotheses, if borne out experimentally, will lead to a greater understanding of shell gland function including the processes involved in eggshell formation and anti-microbial activity. PMID- 19303880 TI - Cloning, distribution and effects of season and nutritional status on the expression of neuropeptide Y (NPY), cocaine and amphetamine regulated transcript (CART) and cholecystokinin (CCK) in winter flounder (Pseudopleuronectes americanus). AB - cDNAs encoding for neuropeptide Y (NPY), cocaine and amphetamine regulated transcript (CART) and cholecystokinin (CCK) were cloned in winter flounder, a species that undergoes a period of natural fasting during the winter. Tissue distribution studies show that these peptides are present in several peripheral tissues, including gut and gonads, as well as within the brain. We assessed the effects of season and fasting on the expression of these peptides. Our results show that NPY and CCK, but not CART, show seasonal differences in expression with higher hypothalamic NPY and lower gut CCK expression levels in the winter. In the summer, fasting induced an increase in hypothalamic NPY expression levels and a decrease in gut CCK levels, but did not affect hypothalamic CART expression levels. None of the peptides examined was affected by fasting in the winter. Our results suggest that NPY and CCK, but maybe not CART, might have a major role in the regulation of feeding in winter flounder and might contribute to the seasonal fluctuations in appetite in this species. PMID- 19303881 TI - Attractive men induce testosterone and cortisol release in women. AB - Recently, Roney et al. (Roney, J.R., Lukaszewski, A.W., Simmons, Z.L., 2007. Rapid endocrine responses of young men to social interactions with young women. Horm. Behav. 52, 326-33; Roney, J.R., Mahler, S.V., Maestripieri, D., 2003. Behavioral and hormonal responses of men to brief interactions with women. Evol. Hum. Behav. 24, 365-375) demonstrated that men release testosterone and cortisol in response to brief social interactions with young women. The current experiment examined whether women show a similar endocrine response to physically and behaviorally attractive men. 120 women (70 naturally-cycling and 50 using hormonal contraceptives) were shown one of four 20-minute video montages extracted from popular films, depicting the following scenarios: 1) an attractive man courting a young woman (experimental stimulus), 2) a nature documentary (video clip control), 3) an unattractive older man courting a woman (male control), and 4) an attractive woman with no men present (female control). Saliva samples were taken before and after presentation of the stimulus, and were later analyzed for testosterone and cortisol content via enzyme immunoassay. Naturally cycling women experienced a significant increase in both testosterone and cortisol in response to the experimental stimulus but to none of the control stimuli. Participants taking hormonal contraceptives also showed a significant cortisol response to the attractive man. Women may release adrenal steroid hormones to facilitate courtship interactions with high mate-value men. PMID- 19303883 TI - Role of Mg(2+) block of the inward rectifier K(+) current in cardiac repolarization reserve: A quantitative simulation. AB - Different K(+) currents serve as "repolarization reserve" or a redundant repolarizing mechanism that protects against excessive prolongation of the cardiac action potential and therefore arrhythmia. Impairment of the inward rectifier K(+) current (I(K1)) has been implicated in the pathogenesis of cardiac arrhythmias. The characteristics of I(K1) reflect the kinetics of channel block by intracellular cations, primarily spermine (a polyamine) and Mg(2+), whose cellular levels may vary under various pathological conditions. However, the relevance of endogenous I(K1) blockers to the repolarization reserve is still not fully understood in detail. Here we used a mathematical model of a cardiac ventricular myocyte which quantitatively reproduces the dynamics of I(K1) block to examine the effects of the intracellular spermine and Mg(2+) concentrations, through modifying I(K1), on the action potential repolarization. Our simulation indicated that an I(K1) transient caused by relief of Mg(2+) block flows during early phase 3. Increases in the intracellular spermine/Mg(2+) concentration, or decreases in the intracellular Mg(2+) concentration, to levels outside their normal ranges prolonged action potential duration by decreasing the I(K1) transient. Moreover, reducing both the rapidly activating delayed rectifier current (I(Kr)) and the I(K1) transient caused a marked retardation of repolarization and early afterdepolarization because they overlap in the voltage range at which they flow. Our results indicate that the I(K1) transient caused by relief of Mg(2+) block is an important repolarizing current, especially when I(Kr) is reduced, and that abnormal intracellular free spermine/Mg(2+) concentrations may be a missing risk factor for malignant arrhythmias in I(Kr) related acquired (drug-induced) and congenital long QT syndromes. PMID- 19303882 TI - Donepezil treatment restores the ability of estradiol to enhance cognitive performance in aged rats: evidence for the cholinergic basis of the critical period hypothesis. AB - Recent studies suggest that the ability of estradiol to enhance cognitive performance diminishes with age and/or time following loss of ovarian function. We hypothesize that this is due, in part, to a decrease in basal forebrain cholinergic function. This study tested whether donepezil, a cholinesterase inhibitor, could restore estradiol effects on cognitive performance in aged rats that had been ovariectomized as young adults. Rats were ovariectomized at 3 months of age, and then trained on a delayed matching to position (DMP) T-maze task, followed by a configural association (CA) operant condition task, beginning at 12-17 or 22-27 months of age. Three weeks prior to testing, rats started to receive either donepezil or vehicle. After one week, half of each group also began receiving estradiol. Acclimation and testing began seven days later and treatment continued throughout testing. Estradiol alone significantly enhanced DMP acquisition in middle-aged rats, but not in aged rats. Donepezil alone had no effect on DMP acquisition in either age group; however, donepezil treatment restored the ability of estradiol to enhance DMP acquisition in aged rats. This effect was due largely to a reduction in the predisposition to adopt a persistent turn strategy during acquisition. These same treatments did not affect acquisition of the CA task in middle-aged rats, but did have small but significant effects on response time in aged rats. The data are consistent with the idea that estrogen effects on cognitive performance are task specific, and that deficits in basal forebrain cholinergic function are responsible for the loss of estradiol effect on DMP acquisition in aged ovariectomized rats. In addition, the data suggest that enhancing cholinergic function pharmacologically can restore the ability of estradiol to enhance acquisition of the DMP task in very old rats following long periods of hormone deprivation. Whether donepezil has similar restorative effects on other estrogen-sensitive tasks needs to be explored. PMID- 19303884 TI - Structural bases for 16 S rRNA methylation catalyzed by ArmA and RmtB methyltransferases. AB - Aminoglycosides are used extensively for the treatment of severe infections due to Gram-negative bacteria. However, certain species have become highly resistant after acquisition of genes for methyltransferases which catalyze post transcriptional methylation of N7-G1405 in 16 S rRNA of 30 S ribosomal subunits. Inactivation of this enzymatic activity is therefore an important challenge for development of an effective therapy. The present work describes the crystallographic structures of methyltransferases RmtB and ArmA from clinical isolates. Together with biochemical experiments, the 3D structures indicate that the N-terminal domain specific for this family of methyltransferases is required for enzymatic activity. Site-directed mutagenesis has enabled important residues for catalysis and RNA binding to be identified. These high-resolution structures should underpin the design of potential inhibitors of these enzymes, which could be used to restore the activity of aminoglycosides against resistant pathogens. PMID- 19303885 TI - SMAR1 forms a ternary complex with p53-MDM2 and negatively regulates p53-mediated transcription. AB - The intra-cellular level of tumor suppressor protein p53 is tightly controlled by an autoregulatory feedback loop between the protein and its negative regulator MDM2. The role of MDM2 in down-regulating the p53 response in unstressed conditions and in the post-stress recovery phase is well documented. However, interplay between the N-terminal phosphorylations and C-terminal acetylations of p53 in this context remains unclear. Here, we show that an MAR binding protein SMAR1 interacts with MDM2 and the Ser15 phosphorylated form of p53, forming a ternary complex in the post stress-recovery phase. This triple complex formation between p53, MDM2 and SMAR1 results in recruitment of HDAC1 to deacetylate p53. The deacetylated p53 binds poorly to the target promoter (p21), which results in switching off the p53 response, essential for re-entry into the cell cycle. Interestingly, the knock-down of SMAR1 using siRNA leads to a prolonged cell cycle arrest in the post stress recovery phase due to ablation of p53-MDM2-HDAC1 interaction. Thus, the results presented here for the first time highlight the role of SMAR1 in masking the active phosphorylation site of p53, enabling the deacetylation of p53 by HDAC1-MDM2 complex, thereby regulating the p53 transcriptional response during stress rescue. PMID- 19303886 TI - Desynchronized vasomotion and desynchronized fiber activation pattern enhance oxygenation in a model of skeletal muscle. AB - Although the full physiological significance of vasomotion is still debated, it is generally thought to have a role in optimizing tissue oxygenation parameters. We study the effect of vasomotion rhythm in skeletal muscle on oxygen transport using a computational model. The model is used: (i) to test a novel hypothesis that "vasomotors" form a chemical network in which the rhythm adapts to meet oxygen demand in skeletal muscle and (ii) to study the contribution of desynchronized/chaotic vasomotion in optimizing oxygen delivery to skeletal muscle. We formulate a 2D grid model of skeletal muscle consisting of an interleaved arrangement of vessels and muscle fibers fired by a motor neuronal network. The vasomotors too form a network interacting by chemical means. When positive (negative) synapses dominate, the neuronal network exhibits synchronized (desynchronized) activity. Similarly, when positive (negative) chemical interactions dominate, the vessels exhibit synchronized (desynchronized) activity. Optimal oxygenation is observed when both neuronal network and vasomotor network exhibit desynchronous activity. Muscle oxygenation is thought to result by interactions between the fiber/neuron network and the vessel network; optimal oxygenation depends on precise rhythm-related conditions on the two networks. The model provides interesting insights into the phenomenon of muscle fatigue. PMID- 19303887 TI - The relationship between dietary energy density and energy intake. AB - Much of the research in ingestive behavior has focused on the macronutrient composition of foods; however, these studies are incomplete, or could be misleading, if they do not consider the energy density (ED) of the diet under investigation. Lowering the ED (kcal/g) by increasing the volume of preloads without changing macronutrient content can enhance satiety and reduce subsequent energy intake at a meal. Ad libitum intake or satiation has also been shown to be influenced by ED when the proportions of macronutrients are constant. Since people tend to eat a consistent weight of food, when the ED of the available foods is reduced, energy intake is reduced. The effects of ED have been seen in adults of different weight status, sex, and behavioral characteristics, as well as in 3- to 5-year-old children. The mechanisms underlying the response to variations in ED are not yet well understood and data from controlled studies lasting more than several days are limited. However, both population-based studies and long-term clinical trials indicate that the effects of dietary ED can be persistent. Several clinical trials have shown that reducing the ED of the diet by the addition of water-rich foods such as fruits and vegetables was associated with substantial weight loss even when patients were not told to restrict calories. Since lowering dietary energy density could provide effective strategies for the prevention and treatment of obesity, there is a need for more studies of mechanisms underlying the effect and ways to apply these findings. PMID- 19303888 TI - Rapid acquisition of conditioned flavor preferences in rats. AB - Rats learn to prefer flavors paired with the post-oral effects of glucose. The present study examined how rapidly they acquire this preference. In Experiment 1, food-restricted rats were given repeated three-session training/testing cycles: one 30-min session with a CS+ flavor paired with intragastric (IG) infusion of 16% glucose, another session with a CS- flavor paired with IG water, and a third session with a choice between the flavors with their infusates. The rats preferred the CS+ (69%) in the first choice session, and preference increased across the six cycles to 86%. These data demonstrate that the post-oral reinforcing action of glucose is potent enough to support one-trial learning. In Experiment 2, two groups of rats were trained in the same way, with the CS+ flavor paired with IG infusion of 16% glucose or 7.1% corn oil emulsion, but tests were conducted under extinction conditions, with both CS+ and CS- flavors paired with IG water. Significant preference for the CS+ was acquired more rapidly with glucose (71% CS+ in test 1) than with oil (69% CS+ in test 4). Consistent with previous work, the post-oral stimulation by glucose was more potent than that of isocaloric oil emulsion in conditioning preferences. The last experiment examined the acquisition rate for a flavor-taste conditioned preference. Rats were trained with a CS+ flavor mixed into an 8% fructose + 0.2% saccharin solution and a CS- flavor in 0.2% saccharin. The same three-session training/testing cycles were used, and in the tests the flavors were presented in saccharin. A significant 74% preference for the CS+ flavor was apparent by the second test. Together these studies show that the acquisition of flavor preferences, whether based on flavor-taste or flavor-nutrient associations, can be quite rapid. PMID- 19303889 TI - Metabolic surgery and gut hormones - a review of bariatric entero-humoral modulation. AB - The global pandemic of obesity is increasing. Inappropriate food intake relative to energy expenditure results in increased adiposity. These factors are partly regulated by signals through the gut-brain and adipose-brain axes. Metabolic operations (otherwise known as Bariatric surgery) offer the most effective results for sustained metabolic improvement and weight loss. They modulate a number of gut hormones that constitute the gut-brain axis. This review summarizes the literature to-date reporting the gut hormone changes associated with these operations and their subsequent effects on appetite. Understanding the anatomical differences between each operation and how these can differentially regulate gut hormonal release can provide new treatments and targets for obesity, appetite and metabolic disorders. PMID- 19303890 TI - Bitter-responsive brainstem neurons: characteristics and functions. AB - The sensation that humans describe as "bitter" is evoked by a large group of chemically diverse ligands. Bitter stimuli are avoided by a range of species and elicit reflex rejection, behaviors considered adaptations to the toxicity of many of these compounds. We review novel evidence for neurons that are narrowly tuned to bitter ligands at the initial stages of central processing. These "B-best" neurons in the nucleus of the solitary tract (NST) and parabrachial nucleus (PBN) respond to multiple types of bitter stimuli and exhibit average responses to bitter tastants that are 6-8 times larger than to moderate concentrations of compounds representing other qualities. However, in the PBN B-best units are appreciably activated by intense salt and acid. Neurons broadly sensitive to salts and acids ("AN" neurons) also responded to bitter stimuli. This sensitivity appeared restricted to stronger intensities of ionic bitters, as cycloheximide remained ineffective across concentrations. In addition to chemosensitive profile, B-best neurons were also distinctive with regard to their posterior receptive fields, long latencies, slow firing rates and projection status. Compared to B-best NST cells, those in the PBN received increased convergence from anterior and posterior receptive fields and responded to a greater number of bitter stimuli. We conclude that B-best neurons likely contribute to pathways underlying gaping, aversive hedonic quality and taste coding. The differential responsiveness of B-best and AN neurons to ionic and nonionic bitter ligands also suggests a potential substrate for discrimination within this quality. PMID- 19303891 TI - Trigeminal perception is necessary to localize odors. AB - The human ability to localize odorants has been examined in a number of studies, but the findings are contradictory. In the present study we investigated the human sensitivity and ability to localize hydrogen sulphide (H(2)S), which in low concentrations stimulates the olfactory system selectively, the olfactory trigeminal substance isoamyl acetate (IAA), and the trigeminal substance carbon dioxide (CO(2)). A general requirement for testing of localization was the conscious perception of the applied stimuli by the participants. Using Signal Detection Theory, we determined the human sensitivity in response to stimulation with these substances. Then the subjects' ability to localize the three different substances was tested. We found that humans can detect H(2)S in low concentration (2 ppm) with moderate sensitivity, and possess a high sensitivity in response to stimulation with 8 ppm H(2)S, 17.5% IAA, 50% v/v CO(2). In the localization experiment, subjects could localize neither the low nor the high concentration of H(2)S. In contrast, subjects possessed the ability to localize IAA and CO(2) stimuli. These results clearly demonstrate that humans, in spite of the aware perception, are not able to localize substances which only activate the olfactory system independent of their concentration, but they possess an ability to localize odorants that additionally excite the trigeminal system. PMID- 19303892 TI - Sex-specific spatio-temporal variability in reproductive success promotes the evolution of sex-biased dispersal. AB - Inbreeding depression, asymmetries in costs or benefits of dispersal, and the mating system have been identified as potential factors underlying the evolution of sex-biased dispersal. We use individual-based simulations to explore how the mating system and demographic stochasticity influence the evolution of sex specific dispersal in a metapopulation with females competing over breeding sites, and males over mating opportunities. Comparison of simulation results for random mating with those for a harem system (locally, a single male sires all offspring) reveal that even extreme variance in local male reproductive success (extreme male competition) does not induce male-biased dispersal. The latter evolves if the between-patch variance in reproductive success is larger for males than females. This can emerge due to demographic stochasticity if the habitat patches are small. More generally, members of a group of individuals experiencing higher spatio-temporal variance in fitness expectations may evolve to disperse with greater probability than others. PMID- 19303893 TI - Transcription factor Nrf2 mediates an adaptive response to sulforaphane that protects fibroblasts in vitro against the cytotoxic effects of electrophiles, peroxides and redox-cycling agents. AB - Sulforaphane can stimulate cellular adaptation to redox stressors through transcription factor Nrf2. Using mouse embryonic fibroblasts (MEFs) as a model, we show herein that the normal homeostatic level of glutathione in Nrf2(-/-) MEFs was only 20% of that in their wild-type counterparts. Furthermore, the rate of glutathione synthesis following its acute depletion upon treatment with 3 micromol/l sulforaphane was very substantially lower in Nrf2(-/-) MEFs than in wild-type cells, and the rebound leading to a approximately 1.9-fold increase in glutathione that occurred 12-24 h after Nrf2(+/+) MEFs were treated with sulforaphane was not observed in Nrf2(-/-) fibroblasts. Wild-type MEFs that had been pre-treated for 24 h with 3 micromol/l sulforaphane exhibited between 1.4- and 3.2-fold resistance against thiol-reactive electrophiles, including isothiocyanates, alpha,beta-unsaturated carbonyl compounds (e.g. acrolein), aryl halides and alkene epoxides. Pre-treatment of Nrf2(+/+) MEFs with sulforaphane also protected against hydroperoxides (e.g. cumene hydroperoxide, CuOOH), free radical-generating compounds (e.g. menadione), and genotoxic electrophiles (e.g. chlorambucil). By contrast, Nrf2(-/-) MEFs were typically approximately 50% less tolerant of these agents than wild-type fibroblasts, and sulforaphane pre treatment did not protect the mutant cells against xenobiotics. To test whether Nrf2-mediated up-regulation of glutathione represents the major cytoprotective mechanism stimulated by sulforaphane, 5 micromol/l buthionine sulfoximine (BSO) was used to inhibit glutathione synthesis. In Nrf2(+/+) MEFs pre-treated with sulforaphane, BSO diminished intrinsic resistance and abolished inducible resistance to acrolein, CuOOH and chlorambucil, but not menadione. Thus Nrf2 dependent up-regulation of GSH is the principal mechanism by which sulforaphane pre-treatment induced resistance to acrolein, CuOOH and chlorambucil, but not menadione. PMID- 19303894 TI - Maternal exposure to anti-androgenic compounds, vinclozolin, flutamide and procymidone, has no effects on spermatogenesis and DNA methylation in male rats of subsequent generations. AB - To verify whether anti-androgens cause transgenerational effects on spermatogenesis and DNA methylation in rats, gravid Crl:CD(SD) female rats (4 or 5/group, gestational day (GD) 0=day sperm detected) were intraperitoneally treated with anti-androgenic compounds, such as vinclozolin (100 mg/kg/day), procymidone (100 mg/kg/day), or flutamide (10 mg/kg/day), from GD 8 to GD 15. Testes were collected from F1 male pups at postnatal day (PND) 6 for DNA methylation analysis of the region (210 bp including 7 CpG sites) within the lysophospholipase gene by bisulfite DNA sequencing method. F0 and F1 males underwent the sperm analysis (count, motility and morphology), followed by DNA methylation analysis of the sperm. Remaining F1 males were cohabited with untreated-females to obtain F2 male pups for subsequent DNA methylation analysis of the testes at PND 6. These analyses showed no effects on spermatogenesis and fertility in F1 males of any treatment group. DNA methylation status in testes (F1 and F2 pups at PND 6) or sperms (F1 males at 13 weeks old) of the treatment groups were comparable to the control at all observation points, although DNA methylation rates in testes were slightly lower than those in sperm. In F0 males, no abnormalities in the spermatogenesis, fertility and DNA methylation status of sperm were observed. No transgenerational abnormalities of spermatogenesis and DNA methylation status caused by anti-androgenic compounds were observed. PMID- 19303895 TI - Rapid extraction combined with LC-tandem mass spectrometry (CREM-LC/MS/MS) for the determination of ciguatoxins in ciguateric fish flesh. AB - Ciguatera is a significant food borne disease caused by potent polyether toxins known as ciguatoxins, which accumulate in the flesh of ciguateric fish at risk levels above 0.1 ppb. The management of ciguatera has been hindered by the lack of analytical methods to detect and quantify clinically relevant levels of ciguatoxin in easily prepared crude extracts of fish. Here we report a ciguatoxin rapid extraction method (CREM) that allows the rapid preparation of fish flesh extracts for the detection and quantification of ciguatoxin by gradient reversed phase liquid chromatography-tandem mass spectrometry (LC/MS/MS). CREM-LC/MS/MS delivers a linear response to P-CTX-1 spiked into fish prior to extraction. A similar response was obtained for P-CTX-1 spiked after extraction, indicating >95% extraction efficiency was achieved overall and 85% at the limit of quantification (0.1 ppb). Using this approach, levels >or=0.1 ppb P-CTX-1 could be detected and quantified from an extract of 2g fish flesh, making it suitable as a confirmatory assay for suspect ciguateric carnivorous fish in the Pacific Ocean. The approach is designed to simplify the extraction and analysis of multiple samples per day. PMID- 19303896 TI - First report on toxicity assessment of the Lessepsian migrant pufferfish Lagocephalus sceleratus (Gmelin, 1789) from European waters (Aegean Sea, Greece). AB - According to the current European Union legislative requirements (Regulation 853/2004/EC; Regulation 854/2004/EC, poisonous fish of the family Tetraodontidae and products derived from them must not be placed on the European markets. Following the increased publicity regarding the presence of the pufferfish species Lagocephalus sceleratus in Greek waters, this study was undertaken in order to confirm its toxicity and assess the risk of poisoning in case of accidental consumption. Acidic extracts from tissues of L. sceleratus specimens of different sizes were examined by means of the official mouse bioassay for tetrodotoxin, while some of the extracts were also tested for the presence of Paralytic Shellfish Poisoning (PSP) toxins with a commercial ELISA kit. Toxicity in mice, with symptomatology indicative of tetrodotoxin, was confirmed in a number of samples and indicated a correlation with fish size. Toxicity of certain tissues (liver, gonads, gastrointestinal tract) in larger individuals, expressed as microg/g tetrodotoxin equivalents, was largely above levels required to cause death in human adults. On the other hand, all tested extracts provided a positive reaction in the ELISA test for PSP toxins. This constitutes the first report for presence of toxicity in L. sceleratus caught in European coastal waters. PMID- 19303897 TI - Phenomenology and neurophysiological correlations: two approaches to perception research. AB - This article argues that phenomenological description and neurophysiological correlation complement each other in perception research. Whilst phenomena constitute the material, neuronal mechanisms are indispensable for their explanation. Numerous examples of neurophysiological correlates show that the correlation of phenomenology and neurophysiology is fruitful. Phenomena for which neuronal mechanism have been found include: (in area V1) filling-in of real and artificial scotomata, contour integration, figure-ground segregation by orientation contrast, amodal completion, and motion transparency; (in V2) modal completion, border ownership, surface transparency, and cyclopean perception; (in V3) alignment in dotted contours, and filling-in with dynamic texture; (in V4) colour constancy; (in MT) shape by accretion/deletion, grouping by coherent motion, apparent motion in motion quartets, motion in apertures, and biological motion. Results suggest that in monkey visual cortex, occlusion cues, including stereo depth, are predominantly processed in lower areas, whereas mechanisms for grouping and motion are primarily represented in higher areas. More correlations are likely to emerge as neuroscientists strive for a better understanding of visual perception. The paper concludes with a review of major achievements in visual neuroscience pertinent to the study of the phenomena under consideration. PMID- 19303898 TI - Employees' sleep duration and body mass index: potential confounders. AB - OBJECTIVE: Productivity losses are associated with both employees' sleep and weight problems. Addressing these issues independently may be complicated by a potential link between sleep duration and weight. The mixed results of prior studies, both supporting and refuting an association between sleep duration and weight, may have been subject to missing variable bias. To clarify future strategies for workplace health promotion, possible confounders to the sleep duration/weight relationship were investigated. METHOD: Multivariate models were used to explore the relationship between self-reported average sleep duration and body mass index (BMI) by sequentially adding blocks of demographic, health behavior, work status, physical health, and emotional status variables. Cross sectional data from the 2007 EADS/Augsburg (Germany) cohort follow-up study (n=1163) were used in the analysis. RESULTS: The relationship between average sleep duration and BMI was significant (beta(St)=-0.06, p=0.04) when demographic, health behavior, and work status variables were included. When physical health and emotional status variables were added, the relationship between sleep duration and BMI did not persist. CONCLUSION: The relationship between employees' sleep duration and weight, if present, involves several pathways and potential confounders that should be taken into account when designing workplace health promotion programs. PMID- 19303899 TI - Complete sequence and analysis of the stability functions of pPSX, a vector that allows stable cloning and expression of Streptomycete genes in Escherichia coli K12. AB - The broad host range, cloning and expression vector pPSX has been completely sequenced and analysed. pPSX is 14.7kb in length and contains the fusion of two continuous segments of the parental 34kb, IncW plasmid pR388. pPSX appears to have retained at least three sets of gene/s which contribute in different ways to plasmid stability. The first of these parB, is a known participant in the partitioning of low-copy number plasmids. While the adjoining gene, orf35, has high homology with kfrA, a putative plasmid nucleoid organiser that is often associated with the ParAB family of proteins. The second set of genes; orfs18, 19, 20, whose exact functions are not clear, have homology to the stability operons of both IncW and IncN plasmids. The third is the resolvase, resP, which may resolve plasmid multimers that can lead to plasmid instability. pPSX is a small, stable cloning vector good for cloning and expression of a wide range of genes, including those from streptomycetes. PMID- 19303900 TI - GABAB receptors as drug targets to treat gastroesophageal reflux disease. AB - For many years, acid-suppressive therapy has been at the forefront of treating gastroesophageal reflux disease (GERD), yet despite the advent of the proton pump inhibitors (PPIs) some patients continue to experience persistent GERD symptoms. Therapeutic (non-surgical) options for such patients are currently limited. To tackle this clinical issue, research efforts have begun to focus on 'reflux inhibition' as a potential therapeutic target - i.e. inhibition of transient lower esophageal relaxations (TLESRs), the predominant mechanism of gastroesophageal reflux. Preclinical research has identified a number of drug targets through which TLESRs can be modulated, and the gamma-aminobutyric acid (GABA) type B (GABA(B)) receptor has emerged as one of the most promising. Studies with baclofen, a well-known agonist of this receptor, have demonstrated that reflux inhibition is a valid concept in the clinical setting in that reducing the incidence of TLESRs improves GERD symptoms. But baclofen is associated with significant central nervous system (CNS) side effects, rendering it undesirable for use as a treatment for GERD. Further development work has yielded a number of novel GABA(B) receptor agonists with reduced CNS side effect profiles, and clinical trials are currently being performed with several agents. Compounds that target TLESRs may therefore present a new add-on treatment for patients with persistent GERD symptoms despite PPI therapy. PMID- 19303901 TI - Glycosphingolipids and insulin resistance. AB - Obesity is associated with an increased risk for insulin resistance, a state characterized by impaired responsiveness of liver, muscle and adipose tissue to insulin. One class of lipids involved in the development of insulin resistance are the (glyco)sphingolipids. Ceramide, the most simple sphingolipid, directly inhibits phosphorylation of the insulin signaling mediator Akt/Protein Kinase B. More complex glycosphingolipids, so-called gangliosides, block phosphorylation of the insulin receptor and down-stream signaling, possibly by exclusion of the insulin receptor from specific membrane domains. Pharmacological inhibition of glycosphingolipid synthesis is found to markedly improve insulin sensitivity in rodent models of insulin resistance. Partial glycosphingolipid reduction is well tolerated and may thus offer an attractive new treatment modality for obesity induced insulin resistance and type II diabetes. PMID- 19303902 TI - Connecting lipid droplet biology and the metabolic syndrome. AB - In the recent years, new advances in the biology of lipid droplets led these structures specialized for lipid storage to be considered as new universal intracellular organelles playing active roles in cell physiology. Concomitantly, studies on the pathogenesis of metabolic diseases such as type 2 diabetes or atherosclerosis, associated with ongoing epidemic obesity, have pointed out the importance of lipotoxic effects in metabolic dysfunction, generated by ectopic lipid storage in non-adipose tissues. The purpose of this paper is to establish connections between recent discoveries in lipid droplet biology and novel views in the pathology of the metabolic syndrome. Bringing together the new concepts produced in these two separated fields might show the way towards the definition of innovative strategies to treat metabolic diseases. Particular attention is given to the role of adipocyte-specific proteins that interact with lipid droplets and confer unique functions to adipocyte lipid storage by limiting the spill-over of fatty acids and their lipotoxic effects. PMID- 19303903 TI - Key amino acid residues in the melanocortin-4 receptor for nonpeptide THIQ specific binding and signaling. AB - Melanocortin 4 receptor (MC4R) plays an important role in the regulation of food intake and glucose homeostasis. Synthetic nonpeptide compound N- (3R)-1 4 tetrahydroisoquinolinium-3-ylcarbonyl-(1R)-1-(4-chlorobenzyl)-2-4-cyclohexyl-4 (1H-1,2,4-triazol-1-ylmethyl)piperidin-1-yl-2-oxoethylamine (THIQ) is a potent agonist at MC4R but not at hMC2R. In this study, we utilized two approaches (chimeric receptor and site-directed mutagenesis) to narrow down the key amino acid residues of MC4R responsible for THIQ binding and signaling. Cassette substitutions of the second, third, fourth, fifth, and sixth transmembrane regions (TMs) of the human MC4R (hMC4R) with the homologous regions of hMC2R were constructed. Our results indicate that the cassette substitutions of these TMs of the hMC4R with homologous regions of the hMC2R did not significantly alter THIQ binding affinity and potency except the substitution of the hMC4R TM3, suggesting that the conserved amino acid residues in these TMs of the hMC4R are main potential candidates for THIQ binding and signaling while non conserved residues in TM3 of MC4R may also be involved. Nineteen MC4R mutants were then created, including 13 conserved amino acid residues and 6 non-conserved amino acid residues. Our results indicate that seven conserved residue [E100 (TM2), D122 (TM3), D126 (TM3), F254 (TM6), W258 (TM6), F261 (TM6), H264 (TM6)] are important for THIQ binding and three non-conserved residues [N123 (TM3), I129 (TM3) and S131 (TM3)] are involved in THIQ selectivity. In conclusion, our results suggest that THIQ utilize both conserved and non-conserved amino acid residues for binding and signaling at hMC4R and non conserved residues may be responsible for MC4R selectivity. PMID- 19303905 TI - Autophagy in intracellular bacterial infection. AB - Numerous pathogens have developed the capacity to invade host cells to be protected from components of the systemic immune system. However, once in the host cells they utilize sophisticated strategies to avoid the powerful machinery built by the cells to kill invading pathogens. In the last few years cumulative evidence indicates that autophagy is one of the most remarkable tools of the intracellular host cell defense machinery that bacteria must confront upon cell invasion. However, several pathogens subvert the autophagic pathway and, manipulate this process at the molecular level, as a strategy to establish a persistent infection. In this review we have summarized the interaction between autophagy and different bacterial pathogens including those that take advantage of the host cell autophagy, allowing successful colonization, as well as those microorganisms which are controlled by autophagy as part of the innate surveillance mechanism. PMID- 19303904 TI - Prolonged exposure of cardiac cells to renin plus angiotensinogen reduces intracellular renin in the failing heart. On the role of angiotensin II-AT1 complex internalization. AB - To investigate the influence of prolonged exposure of cardiac cells to renin plus angiotensinogen (Ao) on intracellular renin levels, myocytes were isolated from the ventricle of cardiomyopathic hamsters(TO-2) and incubated in Krebs solution containing renin(128 pmol Ang ml/min) plus Ao (110 pmol Ang I generated by renin to exhaustion) for a period of 24 h. Membrane-bound and intracellular AT1 receptors levels as well as intracellular renin were studied using immunological methods and quantified by flow cytometry. The results indicated: a) intracellular renin levels were higher in the failing heart at an advanced stage of the disease (8 months) than in age-matched controls; b) the intracellular renin levels were significantly reduced in cells exposed to renin (128 pmol Ang I.ml/min) plus angiotensinogen (Ao)(110 pmol Ang I generated by renin to exhaustion) for a period of 24 h; c) incubation of the cardiomyocytes with renin (128 pmol Ang I.ml/min) alone did not reduced the intracellular renin levels; d) the fall of the intracellular renin level was related to the formation of angiotensin II (Ang II) at the surface cell membrane and internalization of the Ang II-AT1 complex because losartan (10(-7) M) added to the incubation medium containing renin plus Ao, blocked the internalization of AT1 and suppressed the decline of the intracellular renin levels; e) no internalization of renin or renin secretion was found in these experiments. IN CONCLUSION: prolonged exposure of cardiac cells to renin plus Ao (24 h) reduced intracellular renin levels through the internalization of Ang II-AT1 complex and inhibition of renin expression. PMID- 19303906 TI - Safety evaluation of the double mutant 5-enol pyruvylshikimate-3-phosphate synthase (2mEPSPS) from maize that confers tolerance to glyphosate herbicide in transgenic plants. AB - Glyphosate tolerance can be conferred by decreasing the herbicide's ability to inhibit the enzyme 5-enol pyruvylshikimate-3-phosphate synthase, which is essential for the biosynthesis of aromatic amino acids in all plants, fungi, and bacteria. Glyphosate tolerance is based upon the expression of the double mutant 5-enol pyruvylshikimate-3-phosphate synthase (2mEPSPS) protein. The 2mEPSPS protein, with a lower binding affinity for glyphosate, is highly resistant to the inhibition by glyphosate and thus allows sufficient enzyme activity for the plants to grow in the presence of herbicides that contain glyphosate. Based on both a review of published literature and experimental studies, the potential safety concerns related to the transgenic 2mEPSPS protein were assessed. The safety evaluation supports that the expressed protein is innocuous. The 2mEPSPS enzyme does not possess any of the properties associated with known toxins or allergens, including a lack of amino acid sequence similarity to known toxins and allergens, a rapid degradation in simulated gastric and intestinal fluids, and no adverse effects in mice after intravenous or oral administration (at 10 or 2000 mg/kg body weight, respectively). In conclusion, there is a reasonable certainty of no harm resulting from the inclusion of the 2mEPSPS protein in human food or in animal feed. PMID- 19303907 TI - An integrated probabilistic framework for cumulative risk assessment of common mechanism chemicals in food: an example with organophosphorus pesticides. AB - This paper presents a framework for integrated probabilistic risk assessment of chemicals in the diet which accounts for the possibility of cumulative exposure to chemicals with a common mechanism of action. Variability between individuals in the population with respect to food consumption, concentrations of chemicals in the consumed foods, food processing habits and sensitivity towards the chemicals is addressed by Monte Carlo simulations. A large number of individuals are simulated, for which the individual exposure (iEXP), the individual critical effect dose (iCED) and the ratio between these values (the individual margin of exposure, iMoE) are calculated by drawing random values for all variable parameters from databases or specified distributions. This results in a population distribution of the iMoE, and the fraction of this distribution below 1 indicates the fraction of the population that may be at risk. Uncertainty in the assessment is treated as a separate dimension by repeating the Monte Carlo simulations many times, each time drawing random values for all uncertain parameters. In this framework, the cumulative exposure to common mechanism chemicals is addressed by incorporation of the relative potency factor (RPF) approach. The framework is demonstrated by the cumulative risk assessment of organophosphorus pesticides (OPs). By going through this example, the various choices and assumptions underlying the cumulative risk assessment are made explicit. The problems faced and the solutions chosen may be more generic than the present example with OPs. This demonstration may help to familiarize risk assessors and risk managers with the somewhat more complex output of probabilistic risk assessment. PMID- 19303908 TI - Correlates between neurological soft signs and saccadic parameters in schizophrenia. AB - OBJECTIVE: Neurological Soft Signs (NSS) and impairments in oculomotor saccadic paradigms are both frequent in patients with schizophrenia but their correlation has never been explored. METHODS: 78 patients with DSM-IV schizophrenia (including 43 non-treated) and 41 matched healthy controls were tested for NSS, and on three saccadic tasks: prosaccades, predictive saccades and memory-guided saccades) using infrared oculometry. We analyzed correlations between NSS scores and latencies in all three tasks, rate of errors in memory-guided saccades, and rate of anticipated predictive saccades. RESULTS: No correlations were found in healthy controls. In the patient group, the NSS total and motor coordination scores were positively correlated with three saccadic variables: the latency of prosaccades (r=0.36, p<0.01 and r=0.36, p<0.01 respectively), of memory-guided saccades (r=0.35, p<0.01 and r=0.32, p<0.05 respectively) and, negative correlations were found, with the rate of anticipated predictive saccades (r= 0.33, p<0.01; r=-0.35, p<0.01 respectively). NSS total, motor coordination and sensory integration scores were correlated to the latency of non-anticipated predictive saccades (r=0.34, p<0.01; r=0.24, p<0.05 and r=0.40, p<0.001 respectively). The NSS total, motor integration and sensory integration scores were correlated with the rate of errors in memory-guided saccades (r=0.38, p<0.01; r=0.37, p<0.01 and r=0.34, p<0.01 respectively). CONCLUSIONS: These results support a common pathological mechanism with partial overlapping neural substrates between NSS and saccades in schizophrenia. PMID- 19303909 TI - Intensity of opiate withdrawal in relation to the 825C>T polymorphism of the G protein beta 3 subunit gene. AB - OBJECTIVES: The intensity of withdrawal in opiate dependence shows a high inter individual variability. The 825C>T polymorphism (rs5443) of the G-protein beta 3 (GNB3) subunit gene has a strong influence on clinical signs of sympathetic activity in cardiac research. This study was carried out in order to test the hypothesis that carriers of the T allele have an increased sympathetic activity in opiate withdrawal. METHODS: Thirty-nine monovalent opiate addicted patients consecutively admitted to a detoxification ward were investigated. The main parameter for sympathetic activity was the pulse rate in the first 3 days after the regular end of gradual methadone reduction. RESULTS: Thirty-three out of 39 patients achieved a drug-free state: 22 carried a T allele (TT, CT), 11 belonged to the CC genotype group. The pulse rate was significantly (p<0.05) raised in the T allele group compared to the CC genotype group on the first 2 days after stopping methadone administration. In addition, about a third of the T allele carriers needed clonidine treatment on the respective days, but only one patient among the 11 CC homozygotes. There was no significant difference between groups in systolic and diastolic blood pressures as well as in subjective withdrawal ratings. CONCLUSION: A group difference regarding pulse rate could be observed in a small sample and despite a higher degree of concomitant clonidine medication in T allele carriers. The failure to detect group differences in blood pressure and self-rated withdrawal symptoms may be attributed to the more complex regulation of blood pressure and the known weak correlation between objective and subjective withdrawal symptoms. PMID- 19303910 TI - Dose-related hyperprolactinemia induced by venlafaxine. PMID- 19303912 TI - Asymmetric spike-timing dependent plasticity of striatal nitric oxide-synthase interneurons. AB - Corticostriatal projections constitute the major inputs to basal ganglia, an ensemble of sub-cortical nuclei involved in the learning of cognitive-motor sequences in response to environmental stimuli. Besides striatal output neurons (medium-sized spiny neurons, MSNs) in charge of the detection of cortical activity, three main classes of interneurons (GABAergic, cholinergic and nitric oxide (NO)-synthase interneurons) tightly regulate the corticostriatal information transfer. Despite the crucial role of NO on neuronal signaling and synaptic plasticity, little is known about corticostriatal synaptic transmission and plasticity at the level of striatal neuronal nitric oxide synthase (nNOS) interneurons. Using a corticostriatal rat brain slice preserving the connections between the somatosensory cortex and the striatal cells, we have explored the synaptic transmission between the cerebral cortex and striatal nNOS interneurons and their capability to develop activity-dependent long-term plasticity based on the quasi-coincident cortical and striatal activities (spike-timing dependent plasticity, STDP). We have observed that cortical pyramidal cells activate monosynaptically and very efficiently the striatal nNOS interneurons. In addition, nNOS interneurons are able to develop strong bidirectional long-term plasticity, following STDP protocols. Indeed, the strength of cortically-evoked response at nNOS interneurons varied as a function of time interval between pre- and postsynaptic activations (Deltat=t(post)-t(pre)). For Deltat<0, excitatory post-synaptic currents (EPSCs) were depressed, peaking at a delay of -25 ms. For Deltat>0, EPSCs depressed for 00 and long-term potentiation (LTP) induced by "late" Deltat>0. PMID- 19303911 TI - Marine pharmacology in 2005-6: Marine compounds with anthelmintic, antibacterial, anticoagulant, antifungal, anti-inflammatory, antimalarial, antiprotozoal, antituberculosis, and antiviral activities; affecting the cardiovascular, immune and nervous systems, and other miscellaneous mechanisms of action. AB - BACKGROUND: The review presents the 2005-2006 peer-reviewed marine pharmacology literature, and follows a similar format to the authors' 1998-2004 reviews. The preclinical pharmacology of chemically characterized marine compounds isolated from marine animals, algae, fungi and bacteria is systematically presented. RESULTS: Anthelmintic, antibacterial, anticoagulant, antifungal, antimalarial, antiprotozoal, antituberculosis and antiviral activities were reported for 78 marine chemicals. Additionally 47 marine compounds were reported to affect the cardiovascular, immune and nervous system as well as possess anti-inflammatory effects. Finally, 58 marine compounds were shown to bind to a variety of molecular targets, and thus could potentially contribute to several pharmacological classes. CONCLUSIONS: Marine pharmacology research during 2005 2006 was truly global in nature, involving investigators from 32 countries, and the United States, and contributed 183 marine chemical leads to the research pipeline aimed at the discovery of novel therapeutic agents. GENERAL SIGNIFICANCE: Continued preclinical and clinical research with marine natural products demonstrating a broad spectrum of pharmacological activity will probably result in novel therapeutic agents for the treatment of multiple disease categories. PMID- 19303913 TI - Phosphorylation of FADD (Fas-associated death domain protein) at serine 194 is increased in the prefrontal cortex of opiate abusers: relation to mitogen activated protein kinase, phosphoprotein enriched in astrocytes of 15 kDa, and Akt signaling pathways involved in neuroplasticity. AB - Fas-associated protein with death domain (FADD) is a multifunctional protein that can induce both apoptotic and non-apoptotic actions. Recently, FADD was found downregulated in the prefrontal cortex of opiate abusers, which suggested an attenuation of Fas death signals in human addicts. Phosphorylation of FADD (Ser194) has been reported to regulate its non-apoptotic activity, which might include the induction of neuroplastic effects in the brain. This postmortem brain study examined the status of phosphorylated (p)-Ser194 FADD and signaling pathways involved in neuroplasticity in the prefrontal cortex (BA 9) of short term (ST) and long-term (LT) heroin or methadone abusers. In these subjects, the content of monomeric p-FADD was significantly increased when compared with that in age-, gender-, and postmortem delay-matched controls (all addicts: 65%, n=26; ST abuse: 51%; n=11; LT abuse: 75%, n=15). Oligomeric p-FADD forms were modestly increased (11%-23%). At the subcellular level, opiate addiction upregulated the expression of monomeric p-FADD in the nucleus (110%) and that of p-oligomers in the cytosol (66%). In LT opiate addicts (but not ST abusers), a pronounced downregulation of p-extracellular signal-regulated kinase (ERK)1/2 (52%) and p-c Jun NH(2)-terminal protein kinase (JNK)1/2 (51%), but not p-p38 mitogen-activated protein kinase (MAPK), was quantified in the prefrontal cortex (total homogenate and subcellular compartments). Similarly, the signaling pathway mediated by p phosphoprotein enriched in astrocytes of 15 kDa (PEA-15) protein and its phosphorylating kinase p-Akt1 was also downregulated in cortical homogenate (43% and 41%, respectively) and cytosolic preparations of chronic opiate addicts. The results indicate that opiate addiction in humans is associated with an altered balance between p-Ser194 FADD (increased) and total FADD (decreased) in brain, which may favor its neuroplastic actions. The interaction between p-FADD (upregulated) and neuronal pathways (downregulated) could play a relevant role in mediating specific forms of structural and behavioral neuroplasticity. PMID- 19303914 TI - Covert word reading induces a late response in the hand motor system of the language dominant hemisphere. AB - Recent work has demonstrated that overt reading influences the excitability of the language-dominant hand motor cortex. However, this effect was related to speech output, whereas results on silent reading have been inconsistent, and have not allowed for systematic investigation of the different stages of word recognition. To investigate a possible modulation of the cortical excitability mediating hand movements through different stages of covert reading, motor evoked potentials (MEP) from hand muscles in right-handed subjects were recorded. We showed a significant increase of the excitability of the hand motor cortex of the dominant hemisphere during late stages of covert word reading, whereas processing of abstract shapes had no effect and covert articulation induced a decrease in hand motor cortex excitability. There was no significant change of MEP amplitudes during earlier stages of covert reading in the dominant hemisphere or in the non dominant hemisphere. Our results demonstrate a functional connection between cortical networks mediating linguistic processing and hand movements without concurrent activation of the motor cortex through overt articulation at late stages of word reading, which have been shown to involve converging activation of classic left frontal language regions. We speculate that the effect reported here is related to a cortical network mediating gestures which are a part of verbal communication. This supports recent theories on language evolution which postulate that language emerged through manual gestures. PMID- 19303915 TI - Predictability of investment behavior from brain information measured by functional near-infrared spectroscopy: a bayesian neural network model. AB - In line with previous studies using fMRI and as is apparent from experimental results, cerebral blood flow (oxygenated hemoglobin (oxyHb) concentration) in the medial prefrontal cortex (MPFC) and orbital cortex (OFC) as is observed with fNIRS (functional near-infrared spectroscopy) is presumed to be closely related to reward prediction and risk prediction as part of decision-making under risk. Results of analysis using a predictive model with a three-layer perceptron revealed that changes in the oxyHb concentration in cerebral blood as indicated by fNIRS observation include information to effectively predict investment behavior. This paper indicates that adding oxyHb concentration at the aforementioned sites in the brain as a predictive factor allows prediction of subjects' investment behavior with a considerable degree of precision. This fact indicates that information provided by fNIRS allows valid analysis of investment behavior and it also suggests a wide-ranging practical applicability for this information like investment assistance using fNIRS. PMID- 19303916 TI - Developmental changes in point-light walker processing during childhood and adolescence: an event-related potential study. AB - To investigate developmental changes in the neural responses to a biological motion stimulus, we measured event-related potentials (ERPs) in 50 children aged from 7 to 14 years, and 10 adults. Two kinds of visual stimuli were presented: a point-light walker (PLW) stimulus and a scrambled point-light walker (sPLW) stimulus as a control. The sPLW stimulus had the same number of point-lights and the same velocity vector of point-lights as the PLW stimulus, but the initial starting positions were randomized. Consistent with previous ERP studies, one positive peak (P1) and two negative peaks (N1 and N2) were observed at around 130, 200 and 330 ms, respectively, in bilateral occipitotemporal regions, in all age groups. The latency of the P1 component was significantly shorter for the PLW than sPLW stimulus in all age groups, whereas the amplitude was significantly larger for the PLW than sPLW stimulus only for the 7-year-old group. The P1 amplitude and N1 latency were linearly decreased with age. The negative amplitudes of both N1 and N2 components of the PLW stimulus were significantly larger than those of the sPLW stimulus in all age groups. P1-N1 amplitude was changed by development, but not N2 amplitude. These results suggest that the intensity (P1) and timing (N1) of early visual processing for the PLW stimulus changed linearly throughout childhood and P1-N1 amplitude at occipitotemporal electrodes and N1 latency in 10-year-olds, but not 11-year-olds, was significantly larger than that in adults. For the amplitudes of the N2 component in response to PLW and sPLW stimuli in 7-8-year-old subjects were not statistically different from those in adults at occipitotemporal electrodes. These results suggest that the neural response to the PLW stimulus has developed by 10 years of age at the occipitotemporal electrode. PMID- 19303917 TI - Neurobiology of migraine. AB - Migraine is a complex disorder of the brain whose mechanisms are only now being unraveled. It is common, disabling and economically costly. The pain suggests an important role of the nociceptive activation, or the perception of activation, of trigeminal cranial, particularly intracranial afferents. Moreover, the involvement of a multi-sensory disturbance that includes light, sound and smells, as well as nausea, suggests the problem may involve central modulation of afferent traffic more broadly. Brain imaging studies in migraine point to the importance of sub-cortical structures in the underlying pathophysiology of the disorder. Migraine may thus be considered an inherited dysfunction of sensory modulatory networks with the dominant disturbance affecting abnormal processing of essentially normal neural traffic. PMID- 19303918 TI - Impaired baroreflex control of renal sympathetic nerve activity in type 1 diabetic mice (OVE26). AB - To investigate the effects of chronic diabetes on baroreflex control of renal sympathetic nerve activity (RSNA), OVE26 diabetic (transgenic mouse line which develops hyperglycemia within the first 3 weeks after birth) and FVB control mice 5-6 months old were studied. Under anesthesia, RSNA in response to sodium nitroprusside (SNP)- and phenylephrine (PE)-induced mean arterial pressure changes (DeltaMAP) were measured. Baroreflex-induced inhibition of RSNA during PE infusion was characterized using the sigmoid logistic function curve. Baroreflex induced excitation of RSNA during SNP infusion was characterized by the RSNA vs. DeltaMAP relationship. Mean arterial pressure (MAP) responses to the left aortic depressor nerve (ADN) stimulation were evaluated. Compared to FVB control, we found in OVE26 mice that (1) RSNA in response to MAP increase during PE infusion was dramatically reduced, as characterized by the maximal gain of the RSNA sigmoid logistic function curve (FVB: -20.0+/-5.1; OVE26: -7.6+/-0.8%/mm Hg, P<0.05); (2) RSNA in response to MAP decrease during SNP infusion was also attenuated (P<0.05); (3) MAP responses to ADN stimulation were reduced (P<0.05). We concluded that chronic diabetes impairs baroreflex control of RSNA in OVE26 diabetic mice. The use of the transgenic OVE26 diabetic mouse model may underlie a foundation for the further understanding of diabetes-induced autonomic neuropathy. PMID- 19303919 TI - Seizure activity involved in the up-regulation of BDNF mRNA expression by activation of central mu opioid receptors. AB - Chemical-induced seizures up-regulated brain-derived neurotrophic factor (BDNF) mRNA expression. Intracerebroventricular (i.c.v.) administration of endogenous opioids preferentially activating mu opioid receptor (MOR) could also increase BDNF mRNA expression. The aim of this study was to determine to what extent i.c.v. administration of synthetic MOR-selective agonists in rats can modulate both seizure activity and up-regulation of BDNF mRNA expression. Effects and potencies of i.c.v. administration of morphine and [D-Ala(2), N-Me-Phe(4), Gly(5) ol]-enkephalin (DAMGO), were directly investigated by scoring behavioral seizures and measuring BDNF mRNA expression. In addition, effects of the opioid receptor antagonist naloxone and antiepileptic drugs, diazepam, phenobarbital, and valproate, on i.c.v. MOR agonist-induced behavioral seizures and up-regulation of BDNF mRNA expression were determined. A single i.c.v. administration of morphine (10-100 microg) or DAMGO (0.15-1.5 microg) dose-dependently elicited behavioral seizures and increased BDNF mRNA expression in the widespread brain regions. However, s.c. administration of MOR agonists neither produced behavioral seizures nor increased BDNF mRNA expression. Pretreatment with naloxone 1 mg/kg significantly reduced behavioral seizure scores and the up-regulation of BDNF mRNA expression elicited by i.c.v. morphine or DAMGO. Similarly, diazepam 10 mg/kg and phenobarbital 40 mg/kg significantly blocked i.c.v. MOR agonist-induced actions. Pretreatment with valproate 300 mg/kg only attenuated behavioral seizures, but it did not affect morphine-induced increase of BDNF mRNA expression. This study provides supporting evidence that seizure activity plays an important role in the up-regulation of BDNF mRNA expression elicited by central MOR activation and that decreased inhibitory action of GABAergic system through the modulation on GABA receptor synaptic function by central MOR activation is involved in its regulation of BDNF mRNA expression. PMID- 19303921 TI - Forward models of inertial loads in weightlessness. AB - In this experiment, we investigated whether the CNS uses internal forward models of inertial loads to maintain the stability of a precision grip when manipulating objects in the absence of gravity. The micro-gravity condition causes profound changes in the profile of tangential constraints at the finger-object interface. In order to assess the ability to predict the micro-gravity-specific variation of inertial loads, we analyzed the grip force adjustments that occurred when naive subjects held an object in a precision grip and performed point-to-point movements under the weightless condition induced by parabolic flight. Such movements typically presented static and dynamic phases, which permitted distinction between a static component of the grip force (measured before the movement) and a dynamic component of the grip force (measured during the movement). The static component tended to gradually decrease across the parabolas, whereas the dynamic component was rapidly modulated with the micro gravity-specific inertial loads. In addition, the amplitude of the modulation significantly correlated with the amplitude of the tangential constraints for the dynamic component. These results strongly support the hypothesis that the internal representation of arm and object dynamics adapts to new gravitational contexts. In addition, the difference in time scales of adaptation of static and dynamic components suggests that they can be processed independently. The prediction of self-induced variation of inertial loads permits fine modulation of grip force, which ensures a stable grip during manipulation of an object in a new environment. PMID- 19303920 TI - Zic1 levels regulate mossy fiber neuron position and axon laterality choice in the ventral brain stem. AB - Pontine gray neurons of the brain stem are a major source of mossy fiber (MF) afferents to granule cells of the cerebellum. Achieving this connectivity involves an early regionalization of pontine gray neuron cell bodies within the brainstem pontine nuclei, as well as establishing the proper ratio of crossed versus uncrossed MF projections to contralateral versus ipsilateral cerebellar territories. Here, we report expression of the transcription factor Zic1 in newly postmitotic pontine gray neurons and present functional experiments in embryonic and postnatal mice that implicate Zic1 levels as a key determinant of pontine neuron cell body position within the pons and axon laterality. Reducing Zic1 levels embryonically via in utero electroporation of short hairpin RNA interference (shRNAi) vectors shifted the postnatal distribution of pontine neurons from caudolateral to rostromedial territories; by contrast, increasing Zic1 levels resulted in the reciprocal shift, with electroporated cells redistributing caudolaterally. Associated with the latter was a change in axon laterality, with a greater proportion of marked projections now targeting the ipsilateral instead of contralateral cerebellum. Zic1 levels in pontine gray neurons, therefore, play an important role in the development of pontocerebellar circuitry. PMID- 19303922 TI - Olfactory detectability of homologous n-alkylbenzenes as reflected by concentration-detection functions in humans. AB - As part of our systematic exploration of chemical determinants for the olfactory potency of vapors towards humans, we measured concentration-detection functions for the odor of the homologous n-alkylbenzenes toluene, ethylbenzene, butylbenzene, hexylbenzene, and octylbenzene. A vapor delivery device based on dynamic olfactometry and calibrated by gas chromatography, served to test groups of 16 to 17 participants. Subjects were young adults from both genders, normosmics, and nonsmokers. Odor functions were tightly modeled by a sigmoid (logistic) function, both at the group and the individual level. Odor detection thresholds (ODTs), defined as the concentration producing a detectability halfway between chance and perfect detection, decreased with alkyl chain length from toluene (79 ppb) to butylbenzene (2.5 ppb), and then increased form butyl to octylbenzene (89 ppb). The "U"-shaped trend of ODTs as a function of alkyl chain length indicated a loss of odor potency beyond a certain molecular size, a phenomenon recently described for chemosensory irritation (chemesthesis) and that will need consideration in structure-activity models of chemosensory potency. Interindividual ODTs' variability for any single odorant amounted to one order of magnitude, in agreement with recent studies of other homologous series but quite smaller than commonly depicted. PMID- 19303923 TI - Modulation of cocaine-induced activity by intracerebral administration of CXCL12. AB - The role of chemokines in immune function is clearly established. Recent evidence suggests that these molecules also play an important role in the central nervous system as modulators of neuronal activity. The chemokine CXCL12 has been identified in several regions of the adult rat brain including the substantia nigra, ventral tegmental area and caudate putamen. CXCR4, a receptor activated by CXCL12, is expressed by dopaminergic neurons in the substantia nigra. The present study tested the effects of intracranial injections of CXCL12 on cocaine-induced locomotion and stereotypic activity in adult male Sprague-Dawley rats. Results demonstrate that intracerebral ventricular administration of CXCL12 (25 ng/4 microl) 15 min prior to cocaine (20 mg/kg intraperitoneal (i.p.)) produced a significant potentiation of both ambulatory and stereotypic activity as compared to cocaine alone. The effects of CXCL12 were blocked by administration of the selective CXCR4 antagonist, AMD 3100. Administration of CXCL12 into specific brain regions was performed to further understand the site of action of CXCL12. Bilateral administration of CXCL12 (25 ng/0.5 microl) into the ventral tegmental area 15 min prior to cocaine (20 mg/kg i.p.) significantly potentiated cocaine induced ambulatory activity, whereas microinjections of CXCL12 into the caudate putamen selectively increased stereotypy. Conversely, administration of CXCL12 into the lateral shell of the nucleus accumbens resulted in an inhibition of cocaine-stimulated ambulatory activity. No alterations in ambulatory or stereotypic activity were observed following CXCL12 administration into the core of the nucleus accumbens. These results demonstrate that CXCL12 can modulate the behavioral effects produced by cocaine in a brain region-specific manner. PMID- 19303924 TI - Transcriptional regulation of human fibroblast growth factor receptor 1 by E2F-1. AB - Overexpression of fibroblast growth factor receptors (FGFRs) has been observed in many types of human tumors; however, the regulatory mechanism of human FGFR expression is still largely unknown. In the present study, we first identified the transcriptional initiation site in the human FGFR 1 gene by 5'-RACE. Furthermore, we show that the expression of human FGFR 1 is regulated by E2F-1. Characterization of the human FGFR 1 promoter demonstrated that two non-consensus E2F binding sequences at positions +4 to +22 and +25 to +43 relative to our identified transcriptional initiation site in the human FGFR 1 gene were critical for E2F-1-mediated transactivation of human FGFR 1 promoter. Mutations of these sites completely abolished the response of human FGFR 1 promoter to E2F-1 as well as E2F-1 binding in electrophoretic mobility-shift assays. Furthermore, chromatin immunoprecipitation assay showed that E2F-1 was able to bind in vivo to the human FGFR 1 promoter. Moreover, human FGFR 1 protein expression was up-regulated by the overexpression of E2F-1, but down-regulated by the overexpression of pRB in situ, suggesting that the expression of human FGFR 1 is regulated by the pRB/E2F pathway. Because disruption of the pRB/E2F pathway is frequently observed in tumor cells, our findings provide valuable information for studying the role of FGFR 1 in tumor progression. PMID- 19303925 TI - Promoter cloning and characterization of the rabbit BK channel beta1 subunit gene. AB - The beta1 subunit of the voltage-dependent and Ca(2+)-activated large-conductance K(+) channel (BK) in mammalian smooth muscle cells (SMCs) plays an important role in regulating smooth muscle tone and is closely linked with a series of smooth muscle tone associated diseases. However, knowledge of the transcriptional regulation of the BK beta1 is still largely unclear. For the first time, we cloned and characterized the full-length genomic sequence of the rabbit BK beta1 containing a 5'-flanking region of 2021 bp. The full-reading frame of the BK beta1 spans ~7.7 kb and is organized into 4 exons and 3 introns. All of the exon/intron junction sequences contain the GT/AG consensus junction sequence. The transcription initiation site (+1G) is located at 447 bp upstream of the translation initiation codon. Bioinformatics analysis indicated that, without any canonical TATA-box, the 5'-flanking region possesses a high GC content and contains a number of putative transcription factor binding sites. 5'-deletion analysis demonstrated that the region of -93/+30 potentially functions as a core promoter region. A gel mobility shift assay and chromatin immunoprecipitation assay revealed that Sp1 specifically interacts with a putative Sp1-binding site ( 91/-85) in vitro and in vivo. Mutation of this site significantly diminished the promoter activities. Over-expression of Sp1 in smooth muscle cells of rabbit sphincter of Oddi enhanced the promoter activities of the BK beta1 in a dose dependent manner. Thus, we suggest that the Sp1-binding site (-91/-85) is essential to the basal transcription of the rabbit BK beta1. Our studies provide a basic knowledge of the transcription regulation of the rabbit BK beta1. PMID- 19303926 TI - Auditory influences on non-auditory cortices. AB - Although responses to auditory stimuli have been extensively examined in the well known regions of auditory cortex, there are numerous reports of acoustic sensitivity in cortical areas that are dominated by other sensory modalities. Whether in 'polysensory' cortex or in visual or somatosensory regions, auditory responses in non-auditory cortex have been described largely in terms of auditory processing. This review takes a different perspective that auditory responses in non-auditory cortex, either through multisensory subthreshold or bimodal processing, provide subtle but consistent expansion of the range of activity of the dominant modality within a given area. Thus, the features of these acoustic responses may have more to do with the subtle adjustment of response gain within a given non-auditory region than the encoding of their tonal properties. PMID- 19303927 TI - The inheritance of pathogenic mitochondrial DNA mutations. AB - Mitochondrial DNA mutations cause disease in >1 in 5000 of the population, and approximately 1 in 200 of the population are asymptomatic carriers of a pathogenic mtDNA mutation. Many patients with these pathogenic mtDNA mutations present with a progressive, disabling neurological syndrome that leads to major disability and premature death. There is currently no effective treatment for mitochondrial disorders, placing great emphasis on preventing the transmission of these diseases. An empiric approach can be used to guide genetic counseling for common mtDNA mutations, but many families transmit rare or unique molecular defects. There is therefore a pressing need to develop techniques to prevent transmission based on a solid understanding of the biological mechanisms. Several recent studies have cast new light on the genetics and cell biology of mtDNA inheritance, but these studies have also raised new controversies. Here we compare and contrast these findings and discuss their relevance for the transmission of human mtDNA diseases. PMID- 19303928 TI - Dexamethasone-containing biodegradable superparamagnetic microparticles for intra articular administration: physicochemical and magnetic properties, in vitro and in vivo drug release. AB - Compared with traditional drug solutions or suspensions, polymeric microparticles represent a valuable means to achieve controlled and prolonged drug delivery into joints, but still suffer from the drawback of limited retention duration in the articular cavity. In this study, our aim was to prepare and characterize magnetic biodegradable microparticles containing dexamethasone acetate (DXM) for intra articular administration. The superparamagnetic properties, which result from the encapsulation of superparamagnetic iron oxide nanoparticles (SPIONs), allow for microparticle retention with an external magnetic field, thus possibly reducing their clearance from the joint. Two molecular weights of poly(lactic-co-glycolic acid) (PLGA) were used, 12 and 19 kDa. The prepared batches were similar in size (around 10 microm), inner morphology, surface morphology, charge (neutral) and superparamagnetic behaviour. The SPION distribution in the microparticles assessed by TEM indicates a homogeneous distribution and the absence of aggregation, an important factor for preserving superparamagnetic properties. DXM release profiles were shown to be quite similar in vitro (ca. 6 days) and in vivo, using a mouse dorsal air pouch model (ca. 5 days). PMID- 19303929 TI - Improved bioavailability of darunavir by use of kappa-carrageenan versus microcrystalline cellulose as pelletisation aid. AB - The aim of this study was to produce pellet formulations containing a high drug load (80%) of the poorly soluble HIV-protease inhibitor darunavir, using wet extrusion/spheronization with kappa-carrageenan or microcrystalline cellulose (MCC) as pelletization aid. Drug release was assessed in vitro by a standardized paddle-dissolution test and in vivo by a single-dose pharmacokinetic study in dogs. Mean dissolution time (MDT) was 78.2+/-3.5 h from MCC pellets (1301+/-301 microm) and 6.1+/-0.7 min from kappa-carrageenan pellets (966+/-136 microm). In contrast to kappa-carrageenan pellets, MCC pellets did not disintegrate and showed a diffusion-controlled drug release. In line with the in vitro findings, the darunavir peak plasma levels and exposure after the administration of a 300 mg dose were more than 60-fold higher when formulated with kappa-carrageenan pellets when compared with MCC pellets, and 10-fold higher after co administration with 10mg/kg of ritonavir. The relative bioavailability of darunavir versus the reference tablet (F(rel)) was 155% with kappa-carrageenan pellets and 2% with MCC pellets without ritonavir, while 78% and 9%, respectively, in presence of ritonavir. In conclusion, when compared with MCC pellets, the bioavailability of darunavir was substantially improved in kappa carrageenan pellets, likely due to their better disintegration behavior. PMID- 19303930 TI - Cerebrospinal fluid beta-glucocerebrosidase activity is reduced in Dementia with Lewy Bodies. AB - The autophagy-lysosomal degradation pathway plays a role in the onset and progression of neurodegenerative diseases. Clinical and genetic studies indicate that mutations of beta-glucocerebrosidase represent genetic risk factors for synucleinopathies, including Parkinson's Disease (PD) and Dementia with Lewy Bodies (DLB). We recently found a decreased activity of lysosomal hydrolases, namely beta-glucocerebrosidase, in cerebrospinal fluid of PD patients. We have thus measured the activity of these enzymes - alpha-mannosidase (EC 3.2.1.24), beta-mannosidase (EC 3.2.1.25), beta-glucocerebrosidase (EC 3.2.1.45), beta galactosidase (EC 3.2.1.23) and beta-hexosaminidase (EC 3.2.1.52) - in cerebrospinal fluid of patients suffering from DLB, Alzheimer's Disease (AD), Fronto-Temporal Dementia (FTD) and controls. Alpha-mannosidase activity showed a marked decrease across all the pathological groups as compared to controls. Conversely, beta-glucocerebrosidase activity was selectively reduced in DLB, further suggesting that this enzyme might specifically be impaired in synucleinopathies. PMID- 19303931 TI - Expression of HBsAg and preS2-S protein in different yeast based system: a comparative analysis. AB - We have expressed both S and preS2-S genes coding for the hepatitis B small (S) and medium (M) proteins, respectively, in different yeast based expression systems and compared the production level of the recombinant proteins. In Saccharomyces cerevisiae, viral genes were expressed under the inducible Gal10/cyc1 and the constitutive PGK promoters using 2mu replicating vectors. We showed that the yield of S protein was higher than M protein under both inducible (14.27 vs 10.9 mg/l) and constitutive (9.18 vs 6.39 mg/l) conditions, respectively. In the methylotrophic yeast Pichia pastoris, the viral genes were expressed in GS115 (Mut(+): Methanol Utilizing) and KM71 (Mut(S): Methanol Utilizing Slow) under the control of the alcohol oxidase promoter (AOX1). In Mut(S) background, both S and preS2-S genes were expressed at higher levels than in Mut(+). In attempt to increase the yield of recombinant viral proteins in S. cerevisiae, we have co-expressed both inducible and constitutive vectors harboring the S or preS2-S genes leading to recombinant strains called UTS (containing pDP/S+pYePIT/S) and UTP (containing pDP/preS2-S+pYePIT/preS2-S). We showed that the recombinant S and preS2-S proteins were successfully detected and the production level reached 18.31 mg/l for the S and 13.22 mg/l for the M proteins. Our comparative study provides evidence that in small scale, S. cerevisiae is more suitable for HBsAg and preS2-S proteins production than P. pastoris under inducible rather than constitutive condition. PMID- 19303932 TI - Utility of surface-supported bilayers in studies of transmembrane helix dimerization. AB - This review focuses on the methods that are available to study transmembrane (TM) helix dimerization in membrane-like environments (either bacterial membranes or lipid bilayers, as mimics of the eukaryotic cellular membrane), with an emphasis on the utility of surface-supported bilayers in such studies. PMID- 19303933 TI - Composite S-layer lipid structures. AB - Designing and utilization of biomimetic membrane systems generated by bottom-up processes is a rapidly growing scientific and engineering field. Elucidation of the supramolecular construction principle of archaeal cell envelopes composed of S-layer stabilized lipid membranes led to new strategies for generating highly stable functional lipid membranes at meso- and macroscopic scale. In this review, we provide a state of the art survey how S-layer proteins, lipids, and polysaccharides may be used as basic building blocks for the assembly of S-layer supported lipid membranes. These biomimetic membrane systems are distinguished by a nanopatterned fluidity, enhanced stability and longevity and thus, provide a dedicated reconstitution matrix for membrane-active peptides and transmembrane proteins. Exciting areas for application of composite S-layer membrane systems concern sensor systems involving specific membrane functions. PMID- 19303934 TI - Interaural temporal and coherence cues jointly contribute to successful sound movement perception and activation of parietal cortex. AB - The perception of movement in the auditory modality requires dynamic changes in the input that reaches the two ears (e.g. sequential changes of interaural time differences; dynamic ITDs). However, it is still unclear as to what extent these temporal cues interact with other interaural cues to determine successful movement perception, and which brain regions are involved in sound movement processing. Here, we presented trains of white-noise bursts containing either static or dynamic ITDs, and we varied parametrically the level of binaural coherence (BC) of both types of stimuli. Behaviorally, we found that movement discrimination sensitivity decreased with decreasing levels of BC. fMRI analyses highlighted a network of temporal, frontal and parietal regions where activity decreased with decreasing BC. Critically, in the intra-parietal sulcus and the supra-marginal gyrus brain activity decreased with decreasing BC, but only for dynamic-ITD sounds (BC by ITD interaction). Thus, these regions activated selectively when the sounds contained both dynamic ITDs and high levels of BC; i.e. when subjects perceived sound movement. We conclude that sound movement perception requires both dynamic changes of the auditory input and effective sound-source localization, and that parietal cortex utilizes interaural temporal and coherence cues for the successful perception of sound movement. PMID- 19303935 TI - Data-driven intensity normalization of PET group comparison studies is superior to global mean normalization. AB - BACKGROUND: Global mean (GM) normalization is one of the most commonly used methods of normalization in PET and SPECT group comparison studies of neurodegenerative disorders. It requires that no between-group GM difference is present, which may be strongly violated in neurodegenerative disorders. Importantly, such GM differences often elude detection due to the large intrinsic variance in absolute values of cerebral blood flow or glucose consumption. Alternative methods of normalization are needed for this type of data. MATERIALS AND METHODS: Two types of simulation were performed using CBF images from 49 controls. Two homogeneous groups of 20 subjects were sampled repeatedly. In one group, cortical CBF was artificially decreased moderately (simulation I) or slightly (simulation II). The other group served as controls. Ratio normalization was performed using five reference regions: (1) Global mean; (2) An unbiased VOI; (3) Data-driven region extraction (Andersson); (4-5) Reference cluster methods (Yakushev et al.). Using voxel-based statistics, it was determined how much of the original signal was detected following each type of normalization. RESULTS: For both simulations, global mean normalization performed poorly, with only a few percent of the original signal recovered. Global mean normalization moreover created artificial increases. In contrast, the data-driven reference cluster method detected 65-95% of the original signal. CONCLUSION: In the present simulation, the reference cluster method was superior to GM normalization. We conclude that the reference cluster method will likely yield more accurate results in the study of patients with early to moderate stage neurodegenerative disorders. PMID- 19303936 TI - When doing nothing is an option: the neural correlates of deciding whether to act or not. AB - The neural basis of intentionally deciding between different response alternatives has been extensively investigated and associated with the rostral cingulate zone (RCZ). However, from daily experience we know that the decision whether to do something is often prior to the decision what to do. This raises the fundamental question whether the decision to act and the decision not to act can be considered as functionally equivalent. Interestingly, in the legal domain such an equivalence is implicitly assumed by punishing crimes of omission. The aim of the current study was to explicitly test this assumption by comparing the neural representation of intentional actions with intentional non-actions. Our results suggest, that weighing whether to act or not involves similar areas of the brain, namely RCZ and dorsolateral prefrontal cortex, independent of the outcome of this decision. This finding strongly supports the assumption that intentionally not acting can be considered as a mode of action. PMID- 19303937 TI - Parietal regions processing visual 3D shape extracted from disparity. AB - Three-dimensional (3D) shape is important for the visual control of grasping and manipulation. We used fMRI to study the processing of 3D shape extracted from disparity in human parietal cortex. Subjects stereoscopically viewed random-line stimuli portraying a 3D structure, a 2D structure in multiple depth planes or a 2D structure in the fixation plane. Subtracting the second from the first condition yields depth-structure sensitive regions and subtracting the third from the second position-in-depth sensitive regions. Two anterior intraparietal sulcus (IPS) regions, the dorsal IPS medial (DIPSM) and the dorsal IPS anterior (DIPSA) regions, were sensitive to depth structure and not to position in depth, while a posterior IPS region, the ventral IPS (VIPS) region, had a mixed sensitivity. All three IPS regions were also sensitive to 2D shape, indicating that they carry full 3D shape information. Finally DIPSM, but not DIPSA was sensitive to a saccade-related task. These results underscore the importance of anterior IPS regions in the processing of 3D shape, in agreement with their proximity to grasping-related regions. Moreover, comparison with the results of Durand, J.B., Nelissen, K., Joly, O., Wardak, C., Todd, J.T., Norman, J.F., Janssen, P., Vanduffel, W., Orban, G.A., 2007. Anterior Regions of Monkey Parietal Cortex Process Visual 3D Shape. Neuron 55, 493-505 obtained in the monkey indicates that DIPSA and DIPSM may represent human homologues for the posterior part of AIP and the adjoining part of LIP respectively. PMID- 19303938 TI - DNA-damaged polyploid cancer cells can reverse to diploidy: an ordered, but little understood, process of genomic reduction [with reference to the previous comments of Forer (2008) and Wheatley (2008a and b)]. PMID- 19303939 TI - Effect of chronic nNOS inhibition on blood pressure, vasoactivity, and arterial wall structure in Wistar rats. AB - While the unequivocal pattern of endothelial nitric oxide (NO) synthase (eNOS) inhibition in cardiovascular control has been recognised, the role of NO produced by neuronal NOS (nNOS) remains unclear. The purpose of the present study was to describe the cardiovascular effects of NO production interference by inhibition of nNOS with 7-nitroindazole (7-NI). Wistar rats (10 weeks old) were used: control and experimental rats were administered 7-NI 10 mg/kg b.w./day in drinking water for 6 weeks. Systolic blood pressure (BP) was measured by the tail cuff plethysmographic method. Isolated thoracic aortas (TAs) were used to study vasomotor activity of the conduit artery in vitro. The BP response of anaesthetised animals was used to follow the cardiovascular-integrated response in vivo. Geometry of the TA was measured after perfusion fixation (120 mm Hg) by light microscopy. Expression of eNOS was measured in the TA by immunoblot analysis. Although 6 weeks of nNOS inhibition did not alter systolic BP, the heart/body weight ratio was decreased. Relaxation of the TA in response to acetylcholine (10(-9)-10(-5)mol/L) was moderately inhibited. However, no difference in the BP hypotensive response after acetylcholine (0.1, 1, 10 microg) was observed. The contraction of TA in response to noradrenaline (10(-10)-10( 5)mol/L), and the BP pressor response to noradrenaline (0.1, 1 microg) was attenuated. The inner diameter of the TA was increased, and the wall thickness, wall cross-sectional area, and wall thickness/inner diameter ratio were decreased. The expression of eNOS in the TA was increased. In summary, cardiac and TA wall hypotrophy, underlined by decreased contractile efficiency, were observed. The results suggested that two constitutive forms of NOS (nNOS, eNOS) likely participate in regulation of cardiovascular tone by different mechanisms. PMID- 19303940 TI - Plasma pre beta-HDL formation is decreased by atorvastatin treatment in type 2 diabetes mellitus: Role of phospholipid transfer protein. AB - Atorvastatin lowers plasma phospholipid transfer protein (PLTP) activity, which stimulates pre-beta-HDL generation in vitro. We determined the effect of atorvastatin on pre-beta-HDL formation and its relation with PLTP activity in type 2 diabetes. METHODS: Plasma pre-beta-HDL formation as well as plasma apo A I, LpA, LpAI:AII, cholesteryl ester transfer protein (CETP) and PLTP activity were measured before and after 30 weeks treatment in 40 patients randomized to atorvastatin 80 mg daily and 41 placebo receiving patients. Pre-beta HDL formation was measured by crossed immunoelectrophoresis under conditions of lecithin:cholesterol acyltransferase (LCAT) inhibition. RESULTS: Plasma pre-beta HDL formation, triglycerides, LDL cholesterol, PLTP activity, and CETP decreased after statin treatment (all P<0.001 vs placebo), whereas HDL cholesterol increased (P<0.005). Plasma apo A-I, LpAI and LpAI:AII remained unchanged compared to placebo. In all patients combined, the changes in pre-beta-HDL formation were independently related to the decrease in plasma triglycerides (beta=0.31; P=0.006) and PLTP activity (beta=0.23; P=0.038), without a contribution of CETP. In the atorvastatin treated patients, the decrease in pre beta-HDL formation tended to be related to the decrease in PLTP activity (beta=0.30, P=0.061) after controlling for decreases in triglycerides (beta=0.22, P=0.22). CONCLUSION: High dose atorvastatin decreases the capacity of plasma to generate pre-beta-HDL particles in type 2 diabetic patients, probably via lowering of plasma PLTP activity and triglycerides. This could contribute to an improvement in the atherogenic lipoprotein profile. PMID- 19303941 TI - The psychosocial impact of epilepsy in older people. AB - Given a prevalence of 0.5-1%, the epilepsies belong to the most common of neuropsychiatric diseases. Many clinicians think of the epilepsies as an entity beginning in early in life. However, the peak incidence of epilepsy in the developed world is now in the age group beyond 60 years. This population has thus an increasing importance; and in this paper we review and summarize the specific aspects of the clinical presentation, etiology, diagnosis, and treatment of older age patients with epilepsy. In particular, we focus on the specific psychosocial impact that epilepsy has on the life of older people and their carers. PMID- 19303942 TI - The contribution of seizures to psychosocial ill-health. AB - Persons with a chronic health condition may be disadvantaged compared to others, though the precise pattern of disadvantage will vary from one condition to another. Persons with epilepsy have been shown to be at increased risk of both psychological morbidity and social disadvantage. Various clinical characteristics of epilepsy have been linked to these psychosocial risks, primary among which is seizure frequency: studies linking seizure frequency to psychosocial ill-health are reviewed here. Given the apparently powerful influence of seizure frequency, it is unsurprising that psychosocial health trajectories in epilepsy are very closely linked to its clinical course-but the relationship is not a completely linear one. Recent research has begun to unravel factors other than seizure frequency which appear promoting or protective of psychosocial ill-health. The need for a more nuanced approach to understanding the causes of psychosocial ill health is highlighted, as is an important distinction between epilepsy as biomedically defined disease and as socially defined illness. PMID- 19303943 TI - The psychosocial impact of epilepsy in adults with an intellectual disability. AB - Epilepsy has a pervasive impact on the quality of life, and thus the psychosocial well-being, of adults with an intellectual disability. Social and economical well being appears to be affected by an increase in restrictions on activities and thus social contact. The population has very high rates of challenging behaviors and of mental illness. It is likely that these have a significant impact on epilepsy management. When populations with and without epilepsy are controlled for level of ability, no difference in prevalence of behavior or mental illness is seen between the populations. Current knowledge is limited on crucial issues such as the long-term effect of seizure disorder on mental health, quality of life, and cognition. PMID- 19303944 TI - Best clinical and research practice in adults with an intellectual disability. AB - Though the prevalence of epilepsies is substantially higher in people with intellectual disability (ID) compared with the general population, little is known of the psychosocial burden of epilepsy and adjustment and their respective determinants in this important population. In most modern societies, adult life with an ID is characterized by diminished self-determination, autonomous function, and life choices. These factors alone are known to be critically linked to the individual's overall quality of life. The task is to identify additional burdens attributable to coexisting epilepsy. This article addresses the significance, for people with ID who have epilepsy, of the many different factors, including seizure or epilepsy-related parameters, antiepileptic medication, coexisting motor and sensory impairments, and psychopathological and behavioral disorders, that can impact their quality of life. Discussion also covers the methodological difficulties in published studies, and, finally, proposals are outlined for future research in this field. PMID- 19303945 TI - Gender-specific psychosocial impact of living with epilepsy. AB - Although many psychosocial issues affect all people living with epilepsy, certain issues either are specific to one gender or have a different prevalence or significance between men and women with epilepsy. Most studies suggest that the incidence of epilepsy is slightly higher in males with epilepsy. Sexual dysfunction is common among men and women with epilepsy and has been related to epilepsy type and treatment. Women living with epilepsy are often prone to increased seizure frequency at certain phases of their menstrual cycles. Hormone replacement therapy in postmenopausal women may worsen seizures. Treatment during pregnancy is often a precarious balancing act between the teratogenic risks of AEDs and the maintenance of maternal seizure control. However, pregnancy registries and other prospective studies have given us invaluable information on how to optimize treatment regimens as well as information about safety of breastfeeding. These gender-specific factors should be a key consideration when counseling and treating patients with epilepsy. PMID- 19303946 TI - Best clinical and research practice in pediatric neurology. AB - Many children with epilepsy experience it as part of a broader disability. The cause is often a developmental, genetic, or early acquired abnormality associated with learning difficulties and impairments in other systems. In addition, it has become increasingly recognized in recent years that even children with so-called idiopathic generalized epilepsies may have specific language or cognitive impairments. The combination of seizures and these factors can form a significant barrier to the child and family in social and emotional terms as well. A child's brain must mature and learn new skills; and a large proportion of childhood epilepsies directly impact on this. The degree of control of the epilepsy can affect cognitive progress; but for many children the underlying cause of the impairment is the significant factor. For all children, access to appropriate clinical, educational, and social services for assessment and therapy is paramount. Specific initiatives in the North West of the United Kingdom are attempting to address service and organizational issues. Current research projects are investigating the impairments children with epilepsy have in addition to having seizures. PMID- 19303947 TI - Epilepsy treatment as a predeterminant of psychosocial ill health. AB - Epilepsy is a chronic disorder with complex effects on social, vocational, physical, and psychological well-being. Patient-oriented research has demonstrated that recurrent seizures have a strong adverse effect on health related quality of life, but also that seizure rate in persons with pharmacoresistant epilepsy has only a modest correlation with quality of life. Although treatment side effects have received limited attention in epilepsy research, available evidence indicates that adverse medication effects may explain more variance in quality of life than any other clinical variable in persons with pharmacoresistant epilepsy. Furthermore, systematic screening for adverse effects has been shown to be associated with significant reduction in subjective toxicity and improvement in quality of life. There has been only limited research on the relative contribution of specific adverse effects to impaired health-related quality of life. The relative importance of reduction of medication burden after resective epilepsy surgery or vagal nerve stimulation has similarly received sparse attention. Existing deficiencies in the available published research present challenges and opportunities to perform further investigations to define and improve best clinical practices in epilepsy care. PMID- 19303948 TI - Expression of nucleosomal protein HMGN1 in the cycling mouse hair follicle. AB - Here we examine the expression pattern of HMGN1, a nucleosome binding protein that affects chromatin structure and activity, in the hair follicle and test whether loss of HMGN1 affects the development or cycling of the follicle. We find that at the onset of hair follicle development, HMGN1 protein is expressed in the epidermal placode and in aggregated dermal fibroblasts. In the adult hair follicle, HMGN1 is specifically expressed in the basal layer of epidermis, in the outer root sheath, in the hair bulb, but not in the inner root sheath and hair shaft. The expression pattern of HMGN1 is very similar to p63, suggesting a role for HMGN1 in the transiently amplifying cells. We also find HMGN1 expression in some, but not all hair follicle stem cells as detected by its colocalization with Nestin and with BrdU label-retaining cells. The appearance of the skin and hair follicle of Hmgn1(-/-) mice was indistinguishable from that of their Hmgn1(+/+) littermates. We found that in the hair follicle the expression of HMGN2 is very similar to HMGN1 suggesting functional redundancy between these closely related HMGN variants. PMID- 19303949 TI - Are fentanyl and remifentanil safe opioids for rat brain mitochondrial bioenergetics? AB - Fentanyl and remifentanil are potent opioid widely used in routine anesthesia procedures. This study evaluates and compares the effects of fentanyl/remifentanil in isolated brain mitochondria bioenergetic status. Fentanyl and remifentanil in clinical concentrations does not interfere with rat brain isolated mitochondria. Do not withstand, fentanyl concentrations >4 microg/mL, induces an impairment of the respiratory chain characterized by a decrease in respiratory control ratio, state 3 and uncoupled respiration. Additionally, membrane potential collapses and ADP/O were reduced. Remifentanil follows the same profile but with effects at higher concentrations (>10 microg/mL). High concentrations of fentanyl and remifentanil interfere with mitochondrial electron chain (complexes III, IV) and on mitochondrial phosphorylation unit (complex V). Mitochondrial permeability transition pore was not induced by both fentanyl and remifentanil in tested concentrations. These data provide the first indication that fentanyl and remifentanil (microg/mL range) alters mitochondrial metabolism. Fentanyl showed a stronger inhibitory effect on mitochondrial bioenergetics. PMID- 19303950 TI - Mitochondrial disorder with OPA1 mutation lacking optic atrophy. AB - OPA1 is highly expressed in retina and optic nerve. OPA1 mutations were first identified in patients with non-syndromic autosomal dominant optic atrophy. Recently, OPA1 mutations were detected in a multisystemic disorder which has optic atrophy as the core clinical feature and multiple mitochondrial DNA (mtDNA) deletions in muscle. We report a patient with a multisystemic disorder and multiple muscle mtDNA deletions, carrying an in-frame deletion in OPA1 in the absence of optic atrophy. This patient provides evidence that optic atrophy is not the main clinical manifestation of OPA1-related disorders. OPA1 analysis should be considered in mitochondrial disorders despite the lack of optic atrophy. PMID- 19303951 TI - Characterization of a novel acylaminoacyl peptidase with hexameric structure and endopeptidase activity. AB - We have overexpressed in E. coli, purified and investigated the kinetic, thermodynamic and biophysical properties of an acylaminoacyl peptidase (AAP), from the thermophile Pyrococcus horikoshii (PhAAP). It was shown that the electrostatic environment of the catalytic site of PhAAP substantially influenced the pH dependence of the specificity rate constant (k(cat)/K(m)). However, 0.3 M NaCl, which depressed the electrostatic effects, simplified the complex pH-rate profile. The rate of formation of the enzyme-substrate complex (k(1)) was obtained from a non-linear Arrhenius plot. The lack of substrate leaving group effects indicated that k(1) is the rate determining step in the catalysis. DSC and CD measurements demonstrated that PhAAP displayed a stable structure in the catalytically competent pH range. It was shown that PhAAP is not just an acylaminoacyl peptidase, but it also has an endopeptidase activity and so differs from the mammalian AAPs. Size exclusion chromatography with PhAAP revealed a hexameric structure, which is unique among the known members of the prolyl oligopeptidase family that includes AAPs and suggests that its cellular function may be different from that of the dimeric AAP also found in the same organism. PMID- 19303952 TI - Plant proteomics. PMID- 19303953 TI - Toll-like receptor 5- and lymphotoxin beta receptor-dependent epithelial Ccl20 expression involves the same NF-kappaB binding site but distinct NF-kappaB pathways and dynamics. AB - Canonical and alternative NF-kappaB pathways depend on distinct NF-kappaB members and regulate expression of different gene subset in inflammatory and steady state conditions, respectively. In intestinal epithelial cells, both pathways control the transcription of the gene coding the CCL20 chemokine. Lymphotoxin beta receptor (LTbetaR) mediates long lasting CCL20 expression whereas Toll-like receptor 5 (TLR5) signals promote inducible and transient activation. Here, we investigated whether the regulation of ccl20 expression involves different promoter sites and NF-kappaB molecules in response to TLR5 and LTbetaR stimulation. In epithelial cells, both stimulation required the same promoter regions, especially the NF-kappaB binding site but involved different NF-kappaB isoforms: p65/p50 and p52/RelB, for TLR5 and LTbetaR-dependent activation, respectively. The dynamic of activation and interaction with CCL20-specific NF kappaB site correlated with gene transcription. Similar Ccl20 expression and NF kappaB activation was found in the small intestine of mice stimulated with TLR5 and LTbetaR agonists. In summary, different NF-kappaB pathways modulate CCL20 transcription by operating on the same NF-kappaB binding site in the same cell type. PMID- 19303954 TI - Recent progress in understanding dialysis-related amyloidosis. AB - Dialysis-Related Amyloidosis (DRA) is a general amyloidosis, which is specifically found in CKD stage 5 patients. DRA causes various osteoarticular lesions in dialysis patients, and therefore it is not practical to regard this condition separately from chronic kidney disease-mineral and bone disorder (CKD MBD), at least from the viewpoint of daily clinical practice. However, it is still controversial whether this disease condition should be included in CKD-MBD. Recently, a better understanding of the pathogenesis of DRA has been obtained by examination of beta(2)-microglobulin-related amyloid fibril formation, extension, and depolymerization in vitro. Apoliprotein E, proteoglycans, and glycosaminoglycans stabilize the amyloid fibrils. In addition, some lysophospholipids and non-esterified fatty acids accelerate amyloid fibril formation and extension under physiological conditions in vitro. Those molecules may enhance the amyloid deposition in vivo. The frequency and severity of osteoarticular disorders that may be associated with DRA accelerate with the duration of dialysis therapy. We have shown that patients undergoing dialysis therapy for 30 years or more survive with serious complications from osteoarticular disorders. DRA is one of the most harmful osteoarticular complications with regard to the maintenance of daily activities and quality of life in patients undergoing long-term dialysis therapy, in addition to the classical complications of CKD-MBD. PMID- 19303955 TI - The relation of femoral osteon geometry to age, sex, height and weight. AB - As computational modeling becomes an increasingly common tool for probing the regulation of bone remodeling, the need for experimental data to refine and validate such models also grows. For example, van Oers et al. (R.F. van Oers, R. Ruimerman, B. van Rietbergen, P.A. Hilbers, R. Huiskes, Relating osteon diameter to strain. Bone 2008;43: 476-482.) recently described a mechanism by which osteon size may be regulated (inversely) by strain. Empirical data supporting this relation, particularly in humans, are sparse. Therefore, we sought to determine if there is a link between body weight (the only measure related to loading available for a cadaveric population) and osteon geometry in human bone. We hypothesized that after controlling for age, sex and height, weight would be inversely related to femoral osteon size (area, On.Ar; diameter, On.Dm). Secondarily we sought to describe the relation between osteon circularity (On.Cr) and these parameters. Osteons (n=12,690) were mapped within microradiographs of femoral mid-diaphyseal specimens (n=88; 45 male, 43 female; 17-97 yrs). Univariate analysis of covariance was conducted (n=87; 1 outlier) with sex as a fixed factor and height, weight and log-transformed age as covariates. Weight was negatively related to On.Ar and On.Dm (p=0.006 and p=0.004, respectively). Age was significantly related to osteon and, it was also significantly related to circularity (all p<0.001). This relation was negative for On.Ar and On.Dm and positive for On.Cr (increasing circularity with age). On.Ar and On.Dm were found to be significantly different between the sexes (p=0.021 and p=0.019, respectively), with females having smaller osteons. No relation between sex and On.Cr was detected (p=0.449). Height was not significantly related to any of the geometric parameters. Partial eta-squared values revealed that age accounted for the largest proportion (On.Ar: 28%, On.Dm: 18%, On.Cr: 30%), weight accounted for the second largest (On.Ar: 9%, On.Dm: 10%) and sex accounted for the smallest proportion (On.Ar: 6%, On.Dm: 7%) of the variance in geometry. While previous studies have reported relations between osteon size and sex/age, we believe that our findings are the first to demonstrate a link with weight. We believe that this negative relation with weight is most probably mechanical in nature; however, alternative (endocrine) links between bone and adipose tissue cannot be ruled out by our design. PMID- 19303956 TI - The effects of anti-resorptive therapies and estrogen withdrawal in adult scoliosis measured by sub-segmental vertebral BMD analysis. AB - The sub-segmental analysis of dual energy X-ray absorptiometry (DXA) scans from scoliotic vertebrae has established that there are differences in bone mass between the concave and convex sides of the vertebrae. Furthermore, these differences persisted in patients with low bone mass and were related to the geometry and applied loads, suggesting that this is a good model of bone adaptation in response to external stimuli. The goal of this study was to characterize the response of the human scoliotic spine to anti-resorptive treatments and estrogen withdrawal on the concave and convex sides of the spine. A total of 576 vertebrae (199 no treatment, 214 bisphosphonate, 69 estrogen and 94 estrogen withdrawal) were analyzed from 167 postmenopausal, Caucasian women. An analysis of variance (ANOVA) was used in conjunction with post-hoc Tukey tests to examine the effects of concavity, treatment group, and age. We found that the average change in BMD per year was greater than zero on the concave and convex sides with the exception of the estrogen withdrawal group. Discontinuing estrogen therapy caused patients to maintain bone mass on the concave side, but lose substantial bone density on the convex side. A differential response was also observed with respect to age. Patients younger than 60 exhibited a decrease in total BMD per year concomitant with a small degree of straightening, while those who were 60 or over demonstrated an increase in bone mass and a slight increase in the deformity. Based on these data, it is clear that the differences in BMD between the concave and convex sides of the vertebrae are not simply a result of the deformity, but more likely due to bone accretion. Further study is needed to elucidate the relationship between biomechanical forces and the adaptive response in the spine as a function of time. PMID- 19303957 TI - Effects of calcimimetic on vascular calcification and atherosclerosis in uremic mice. AB - Recent in vitro and in vivo studies suggest that the calcium-sensing receptor (CaR) plays a role in the process of vascular calcification. Whether it is also involved in the process of atherosclerosis remains an open issue. It is of interest to note that CaR expression is reduced in the arteries of uremic patients, compared with that of non-uremic subjects, and that the progression of vascular calcification in uremic patients is much faster than in general population. It is therefore important to identify treatments which allow us to slow this rapid progression. In this context, it was tempting to examine possible vascular effects of a calcimimetic in the setting of chronic kidney disease (CKD), all the more since cinacalcet, an allosteric modulator of the CaR, has been recently approved for the treatment of hyperparathyroidism secondary to CKD. We have therefore tested the effects of the calcimimetic R-568 in the experimental model of the uremic apoE(-/-) mouse. We have been able to demonstrate that this calcimimetic did not only delay the progression of aortic calcification, but also that of atherosclerosis. This beneficial effect might have occurred through systemic as well as direct local effects, probably via an activation of the CaR in vascular endothelial and smooth muscle cells. The present review is therefore devoted to the effects of calcimimetics on uremia induced vascular disease. PMID- 19303961 TI - Influence of obesity on vitamin D-binding protein and 25-hydroxy vitamin D levels in African American and white women. AB - 25-Hydroxy vitamin D (25OHD) is lipophilic and highly bound to vitamin D-binding protein (VDBP) in plasma. In the present study, we examined VDBP and 25OHD levels by race and body mass index (BMI) in young adult women to determine whether circulating VDBP plays a role in the low levels of 25OHD with obesity and among African Americans. In agreement with previous studies, mean 25OHD levels were lower in African American women than in whites (P < .01). In a hierarchical multiple regression model, BMI was associated with 25OHD after adjustment for age in white women (P = .02, R(2) = .10) but not in African American women. The VDBP levels, by contrast, were similar in African Americans and whites, and were unrelated to BMI in either racial group. Furthermore, VDBP was unrelated to the plasma level of 25OHD. These data confirm an interaction between race and obesity in vitamin D metabolism, and imply that the carrier protein is not an important determinant of circulating 25OHD in women, nor is it affected by race or adiposity. PMID- 19303962 TI - Proinflammatory cytokines in response to insulin-induced hypoglycemic stress in healthy subjects. AB - Hyperglycemic crises of diabetic ketoacidosis and nonketotic hyperglycemia are associated with elevation of counterregulatory hormones and proinflammatory cytokines, markers of lipid peroxidation, and oxidative stress. To investigate if other conditions besides hyperglycemia could evoke such a prompt increase in cytokine levels, lipid peroxidation, and oxidative stress markers, we induced hypoglycemic stress by standard insulin tolerance test and measured proinflammatory cytokines, markers of lipid peroxidation, reactive oxygen species (ROS), and counterregulatory hormones. Insulin tolerance test was performed in 13 healthy male subjects with no history of infection, cardiovascular risk factors, or abnormal glucose. At baseline and at 30, 45, 60, 120, and 240 minutes after insulin injection, the following parameters were measured: glucose, cortisol, corticotropin, epinephrine (EP), norepinephrine (NE), growth hormone, tumor necrosis factor (TNF)-alpha, interleukin (IL) 1beta, IL-6, IL-8, free fatty acids, white blood cells, lipid peroxidation markers by thiobarbituric acid assay, and ROS by dichlorofluorescein method. The peak value of white blood cell count at 120 minutes was significantly associated with the peak values of NE at 30 minutes and cortisol at 60 minutes. By comparing the area under the curve of measured parameters, EP emerged as significant predictor of TNF-alpha (P = .05) and IL-8 (P = .027). Cortisol emerged as predictor of IL-1beta significantly (P = .05). Corticotropin predicted area under the curve of IL-6 with borderline significance (P = .06). In the present study, insulin-induced hypoglycemia in nondiabetic male subjects is associated with increased proinflammatory cytokines (TNF-alpha, IL-1beta, IL-6, and IL-8), markers of lipid peroxidation, ROS, and leukocytosis. Elevations of NE, EP, corticotropin, and cortisol in hypoglycaemia are associated with the elevation of the proinflammatory cytokines and leukocytosis. PMID- 19303963 TI - Preoperative glucocorticoid administration attenuates the systemic stress response and hyperglycemia after surgical trauma in the rat. AB - The stress response to surgery is characterized by activation of the hypothalamic pituitary-adrenal axis and the sympathetic nervous system, and by an inflammatory response and hyperglycemia. The aim of the present study was to investigate if preoperative corticosterone could reduce the postoperative systemic stress response, without aggravating hyperglycemia or interfering with activation of the hypothalamic-pituitary-adrenal axis, in a standardized rat model of surgical trauma. We used a standardized experimental model of intestinal resection in the rat. Exogenous corticosterone (8 mg/kg body weight) or vehicle was administered 2 hours before surgery; and postoperative plasma concentrations of interleukin-6, interleukin-10, adrenaline, noradrenaline, glucose, and insulin were determined. Exogenous corticosterone decreased preoperative plasma adrenaline but did not change plasma glucose or insulin levels. Moreover, corticosterone reduced postoperative plasma interleukin-6, catecholamines, and glucose (all P < .001 .05) without any effect on the plasma corticosterone concentration compared with vehicle-treated controls. A preoperative 2-hour exposure of physiologic poststress corticosterone concentrations not only suppressed plasma IL-6 levels but also inhibited surgery-induced adrenaline release and suppressed plasma glucose levels. We hypothesize that glucocorticoids attenuated the inflammatory response in injured tissues that reduced afferent input into brain areas regulating the neuroendocrine response. PMID- 19303964 TI - Middle-aged Japanese women are resistant to obesity-related metabolic abnormalities. AB - We attempted to determine sex differences in obesity-related metabolic abnormalities in a relatively large middle-aged Japanese population. The study population consisted of 2935 men and 1622 women who were 35 to 59 years old. Metabolic abnormalities were determined using the Japanese criteria for metabolic syndrome, and we evaluated the number of metabolic abnormalities discriminated by waist circumference. In men, the mean number of metabolic abnormalities increased as the waist circumference increased. In women, although the mean number of metabolic abnormalities increased as the waist circumference increased, the mean number was less than 1 even in those with a waist circumference of at least 95 cm. According to the receiver operating characteristic curve, the cutoff levels yielding the maximal sensitivity plus specificity for predicting the prevalence of one or more obesity-related metabolic abnormalities were 80 cm in men and 73 cm in women. However, the positive predictive value was as low as 28.8% in men and 7.1% in women, which may not be suitable for a screening test, especially in women. Middle-aged Japanese women seem to be resistant to obesity-induced metabolic abnormalities, and waist circumference would not effectively predict the existence of metabolic syndrome. In setting the cutoff points in guidelines, a greater emphasis should be placed on the absolute risk of having abnormalities or diseases. PMID- 19303965 TI - Fruit and vegetable consumption and risk factors for cardiovascular disease. AB - The international guidelines issued by the World Health Organization recommend reduction in dietary saturated fat and cholesterol intakes as means to prevent hypercholesterolemia and cardiovascular disease (CVD); however, only limited data are available on the benefits of fruit and vegetable consumption on CVD risk factors in a community-based population. The aim of this study was to examine whether, and to what extent, intake of fruits and vegetables is inversely associated with CVD risk factors in adults. In this population-based cross sectional study, a representative sample of 840 Tehranian adults (male and female) aged 18 to 74 years was randomly selected in 1998. Multivariate logistic regression adjusted for lifestyle and nutritional confounders was used in 2 models. After adjusting for confounders, dietary fruit and vegetable were found to be significantly and inversely associated with CVD risk factors. Adjusted odds ratio for high low-density lipoprotein concentrations were 1.00, 0.88, 0.81, and 0.75 (P for trend < .01) in the first model, which was adjusted for age, sex, keys score, body mass index, energy intake, smoking status, dietary cholesterol, and history of diabetes mellitus and coronary artery disease, a trend which was not appreciably altered by additional adjustment for education, physical activity, and saturated, polyunsaturated, and total fat intakes. This association was observed across categories of smoking status, physical activity, and tertiles of the Keys score. Exclusion of subjects with prevalent diabetes mellitus or coronary artery disease did not alter these results significantly. Consumption of fruits and vegetables is associated with lower concentrations of total and low density lipoprotein cholesterol and with the risk of CVD per se in a dose response manner. PMID- 19303966 TI - The response of gamma vitamin E to varying dosages of alpha vitamin E plus vitamin C. AB - Vitamin E has been studied extensively in the prevention of atherosclerosis. Cross-sectional population studies as well as randomized controlled intervention trials have demonstrated conflicting results. A recent meta-analysis of these trials has emphasized the ineffectiveness of vitamin E in atherosclerosis prevention, with a possibility of harm at higher dosages. However, vitamin E has several isomers, with the alpha form being available via dietary supplements and the gamma form being available via dietary foodstuffs. The gamma form of vitamin E demonstrates several superior properties (such as trapping reactive nitrogen species and detoxifying nitrogen dioxide) compared with alpha vitamin E. All clinical trials have used the alpha isomer, with little concern that this isomer of vitamin E may actually suppress the gamma isomer of vitamin E. We undertook a dose-response study in volunteers with type 2 diabetes mellitus to include all the dosages of alpha vitamin E that have been used in cardiovascular prevention trials to determine the effect of alpha vitamin E on gamma vitamin E. We also assessed the effect of alpha vitamin E on several traditional markers of atherosclerotic risk. We added vitamin C to the vitamin E because several clinical trials included this vitamin to enhance the antioxidant effects of alpha vitamin E. Volunteers received, in randomized order for a 2-week period, one of the following vitamin dosage arms: (1) no vitamins, (2) low-dose supplemental vitamins E plus C, (3) medium-dose supplemental vitamins E plus C, and (4) high dose supplemental vitamins E plus C. Blood levels of both alpha and gamma vitamin E were measured as well as surrogate markers of oxidative stress, hypercoagulation, and inflammation during a high-fat atherogenic meal (to increase the ambient oxidative stress level during the study). The results demonstrate that alpha vitamin E levels increased in proportion to the dose administered. However, at every dose of alpha vitamin E, gamma vitamin E concentration was significantly suppressed. No beneficial changes in surrogate markers of atherosclerosis were observed, consistent with the negative results of prospective clinical trials using alpha vitamin E. Our results suggest that all prospective cardiovascular clinical trials that used vitamin E supplementation actually suppressed the beneficial antioxidant gamma isomer of vitamin E. No beneficial effects on several potential cardiovascular risk factors were observed, even when the vitamin E was supplemented with vitamin C. If a standardized preparation of gamma vitamin E (without the alpha isomer) becomes available, the effects of gamma vitamin E on atherosclerotic risk will warrant additional studies. PMID- 19303967 TI - Retinol-binding protein levels are increased in association with gonadotropin levels in healthy women. AB - Recent studies have demonstrated an association between retinol-binding protein (RBP4) and insulin resistance. Retinol-binding protein is decreased in women and elevated in polycystic ovary syndrome. However, prior studies have not investigated the relationship between RBP4, gonadal steroids, and gonadotropins in healthy women. The aim of this study was to determine the RBP4 levels in a cohort of healthy women with a range of body mass indices and glucose tolerances to investigate the relationship between RBP4, gonadotropin levels, and menopausal status. Serum RBP4 levels were measured by enzyme-linked immunosorbent assay and quantitative Western blot in 88 healthy women (aged 24-59 years) from the general community in a cross-sectional study. Retinol-binding protein was higher in postmenopausal compared with premenopausal women (26.1 +/- 2.1 vs 19.3 +/- 0.5 mug/mL, P = .001). In univariate analysis, RBP4 was associated with follicle stimulating hormone (r = 0.37, P = .0004), luteinizing hormone (r = 0.3, P = .005), and sex hormone-binding globulin (r = -0.24, P = .03) and trended to significance with estradiol (P = .09) but not with free testosterone or dehydroepiandrosterone sulfate. Retinol-binding protein was also associated with insulin at 2 hours during an oral glucose tolerance test (r = 0.24, P = .03) and the area under the curve for insulin during the oral glucose tolerance test (r = 0.26, P = .02). In multivariate regression modeling, both follicle-stimulating hormone (P = .03) and luteinizing hormone (P = .04) remained significantly associated with RBP4 after controlling for estradiol, sex hormone-binding globulin, insulin area under the curve, cholesterol, triglycerides, waist-to-hip ratio, and tumor necrosis factor alpha. Retinol-binding protein was not associated with inflammatory markers or with carotid intima-media thickness. Therefore, RBP4 is higher in postmenopausal women and is associated with gonadotropin concentrations in healthy women. PMID- 19303968 TI - Utilization of an abbreviated diabetes impact management scale to assess change in subjective disability during a trial of pulsatile insulin delivery demonstrates benefit. AB - A prospective interventional study of pulsatile intravenous insulin infusion therapy has demonstrated reduction of left ventricular mass and blunting of progression of diabetic nephropathy. We anticipated that improvements in objective parameters would be associated with similar improvement measurable by the self-administered Diabetes Impact Management Scale (DIMS). The DIMS was administered at baseline and 12 months for 19 participants randomized to receive either standard insulin treatment of 3 to 4 injections of insulin daily or insulin treatment plus an additional day per week of 3 intravenous pulses over an 8-hour period. For standard vs pulsed intravenous insulin therapy, mean baseline scores were similar for the 12 total questions as well as the groups of 7 questions with emotional content and 5 with physical (neurologic) content. Mean study group scores at 1 year and changes over 1 year were not significantly different for the 7 questions with emotional content (P = .3143, .7574). Score results for the 5 questions related to neurologic status at 1 year and changes over 1 year were significantly different between patients with standard and with pulsed insulin therapy (P = .0144, 0.0004). Pulsatile intravenous insulin, when added to standard multiple-dose insulin therapy, was demonstrated to improve subjective perception of neurologic disability on repeated use of an abbreviated form of the DIMS. PMID- 19303969 TI - A pilot study to assess utility of changes in elements of the Diabetes Impact Management Scale in evaluating diabetic patients for progressive nephropathy. AB - A prospective study involving the use of the Diabetes Impact Management Scale (DIMS) in individuals with diabetic nephropathy as part of an interventional study of pulsatile intravenous insulin infusion therapy is used to define the utility of repeated subjective DIMS testing. We hypothesized that repeated use of such an evaluation would correlate well with other objective end points. The DIMS was administered at baseline and 12 months for 19 participants randomized to receive either standard insulin treatment of 3 to 4 injections of insulin daily or standard insulin treatment plus an additional day per week of 3 intravenous pulses over an 8-hour period. Measures of glycemic control, renal function, hemostatic factors, hemodynamics, left ventricular mass, and function were assessed at baseline and 12 months. Of 44 questions on impact of diabetes management, only 12 (5 reflecting physical and 7 reflecting emotional status) showed significant change from baseline to 1 year. Changes in the 5 physical questions related to neurologic status correlated with stable creatinine (P = .0001), stable creatinine clearance (P = .0001), and decrease in left ventricular hypertrophy (P =.0117). Repeated use of an abbreviated, standardized subjective instrument uncovered changes in quality of life that correlated with differences in renal function and left ventricular mass over 12 months. Further use of such an instrument may help us focus treatment for maximum impact. PMID- 19303970 TI - Association between leptin and transaminases: 1-year follow-up study in 180 overweight children. AB - Leptin and insulin resistance are being discussed to be involved in the pathogenesis of nonalcoholic fatty liver disease, which is frequently characterized by moderately elevated transaminases. However, longitudinal studies proving an association between leptin, insulin resistance, and transaminases are scarce. We examined weight status, aspartate aminotransferase (AST), alanine aminotransferase (ALT), leptin, glucose, and insulin in 180 overweight children at baseline and 1 year later. Relationships between these parameters at baseline and their changes in the course of 1 year were determined by multiple regression analysis adjusted for age, sex, pubertal stage, and body mass index (BMI). Leptin but not homeostasis model assessment of insulin resistance index correlated significantly to transaminases in both cross-sectional and longitudinal analyses. The same findings were observed in 30 children with suspected nonalcoholic fatty liver disease by ultrasound. The 130 children who participated in a 1-year lifestyle intervention reduced their overweight (standard deviation score [SDS] BMI, -0.37 +/- 0.11). In the course of 1 year, their changes of transaminases depended on change of weight status (SDS-BMI decrease >0.5: ALT 12 [10-15] --> 9 [8-13] U/L, AST 11 [9-12] --> 9 [8-12] U/L; SDS-BMI decrease >0 but 16 [12-26] U/L, AST 10 [8-14] --> 10 [8-24] U/L; no SDS-BMI decrease: ALT 13 [11-20] --> 20[13-33] U/L, AST 11 [9-21] --> 15 [9-24] U/L; data as median and interquartile range). The 50 children without intervention increased their SDS-BMI (+0.02 +/- 0.18) and transaminases (ALT 14 [11-18] --> 19 [15-25] U/L, AST 10 [8-15] --> 16 [10-25] U/L). These findings suggest that leptin may be involved in the pathogenesis of liver diseases. However, to test this hypothesis, careful histologic assessments in correlation to leptin levels are needed. PMID- 19303971 TI - Diabetic nephropathy and risk factors for peripheral artery disease in Chinese with type 2 diabetes mellitus. AB - The risk for peripheral arterial disease (PAD) is increased in patients with chronic kidney disease. We investigated the effects of renal function on PAD in Chinese with type 2 diabetes mellitus. This study enrolled a total of 2983 (1342 men and 1641 women) Chinese adults with diabetes. The mean age was 63.2 +/- 11.9 years. Peripheral arterial disease was diagnosed by an ankle-brachial index less than 0.9. Renal function was evaluated by serum creatinine (SCr), estimated glomerular filtration rate, and urinary albumin-creatinine ratio (ACR). Risk factors for PAD were evaluated using multiple logistic regression analysis. Age, cholesterol, and high-density lipoprotein cholesterol (HDL-C) (inverse association) were significant risk factors in men, whereas age, body mass index (inverse association), low-density lipoprotein cholesterol, and HDL-C (inverse association) were significant risk factors for diabetic women. After adjustment for age, body mass index, blood pressure, glycosylated hemoglobin, cholesterol, HDL-C, low-density lipoprotein cholesterol, and triglyceride levels, we found that SCr levels greater than 1.5 mg/dL, estimated glomerular filtration rate less than 60 mL/min, and urinary ACR greater than 30 mg/g were independent risk factors for PAD in diabetic men and that SCr levels greater than 1.4 mg/dL and urinary ACR greater than 30 mg/g were independently associated with PAD in diabetic women. The risk factors for PAD are somewhat different between men and women with diabetes in Chinese population in Taiwan. Diabetic nephropathy is significantly associated with PAD in this patient population. PMID- 19303972 TI - Beneficial effects of designed dietary fatty acid compositions on lipids in triacylglycerol-rich lipoproteins among Chinese patients with type 2 diabetes mellitus. AB - Elevated levels of postprandial triacylglycerol-rich lipoproteins (ppTRLs) are atherogenic. Patients with type 2 diabetes mellitus (T2DM) have exaggerated postprandial lipemia associated with elevation or prolonged residence of ppTRL remnants. We examined whether dietary fatty acid compositions (DFACs) decrease atherogenic lipid profiles in ppTRL subfractions in T2DM Chinese patients. A single-blind randomized controlled trial was conducted among 28 T2DM patients. Patients consumed 1 of 3 standardized DFAC-specific fat meals: equidominant (1:1:1), polyunsaturated fatty acid (PUFA)-dominant (PUFA-D, 1:1.7:2.3), or monounsaturated fatty acid (MUFA)-dominant (MUFA-D, 1:1.7:1.2) meals. Numbers in parenthesis, respectively, represent the ratio of saturated fatty acids, MUFA, and PUFA to saturated fatty acids. The MUFA-D meal was the control. Triacylglycerol and cholesterol levels were measured in Svedberg flotation rate (S(f)) greater than 400, S(f) 60 to 400, S(f) 20 to 60, and S(f) 12 to 20 ppTRL subfractions at fasting (0 hour) and 2, 4, and 6 hours after the consumption of the fat meals. Effects of DFACs on mean concentrations of triacylglycerols and cholesterol averaged over 0, 2, 4, and 6 hours in ppTRL subfractions were assessed using linear mixed models. Stability and robustness were validated with 1000 bootstrap replicates. Contrasted to the control, equidominant meal reduced 6 hour average triacylglycerol levels in S(f) greater than 400 (P = .002, bootstrap P < .05) and S(f) 20 to 60 (P = .02, bootstrap P < .05) subfractions, and decreased average S(f) 20 to 60 cholesterol (P = .04, bootstrap P < .05); PUFA-D decreased S(f) greater than 400 average triacylglycerol levels (P = .09, bootstrap P < .05). Bootstrap samples suggested that PUFA-D decreased average S(f) 20 to 60 cholesterol levels (bootstrap P < .05). Therefore, modifying DFACs attenuates the atherogenic lipid profile of ppTRLs in T2DM patients; but increasing PUFA ratio may be more feasible. PMID- 19303973 TI - RARRES2, encoding the novel adipokine chemerin, is a genetic determinant of disproportionate regional body fat distribution: a comparative magnetic resonance imaging study. AB - Visceral fat mass is a strong and independent predictor of obesity-related disorders. To date, little is known about the genetic determinants of regional body fat distribution in humans. As candidates of regional fat distribution, we investigated the fat mass- and obesity-associated gene, the peroxisome proliferator-activated receptor-delta gene, and the retinoic acid receptor responder 2 (RARRES2) gene. We studied whether genetic variation within these genes contributes to the development of disproportionate visceral obesity and obesity-related traits, such as insulin resistance and beta-cell dysfunction. We genotyped 337 subjects with an increased risk for type 2 diabetes mellitus for tagging single nucleotide polymorphisms (SNPs) in the 3 genes and performed association analyses with anthropometric data and parameters of insulin sensitivity and beta-cell function. All subjects underwent an oral glucose tolerance test; a subset was additionally characterized by a hyperinsulinemic euglycemic clamp. Body fat distribution was assessed by nuclear magnetic resonance imaging. The fat mass- and obesity-associated gene SNP rs8050136 was nominally associated with body mass index (P = .0130), but not with body fat distribution, after appropriate adjustment. Magnetic resonance imaging-quantified visceral fat mass was significantly associated with RARRES2 SNP rs17173608 and nominally associated with RARRES2 SNP rs10278590 in nonobese subjects (P = .0002 and P = .0423, respectively), with carriers of the minor alleles displaying lower visceral adipose tissue mass. Besides, the minor allele of SNP rs17173608 was nominally associated with a lower waist-to-hip ratio (P = .0295). In obese subjects, these associations were not detected. No associations were found between the peroxisome proliferator-activated receptor-delta gene and measures of whole-body adiposity and of body fat distribution. All SNPs were associated neither with insulin sensitivity nor with insulin secretion. Common genetic variation within RARRES2 is associated with increased visceral fat mass in nonobese subjects. In generalized obesity, this genetic effect may be masked by the close association between whole-body obesity and visceral fat mass. PMID- 19303974 TI - Metformin suppresses high glucose-induced poly(adenosine diphosphate-ribose) polymerase overactivation in aortic endothelial cells. AB - Overactivation of poly(adenosine diphosphate-ribose) polymerase (PARP), an enzyme involved in cellular response to DNA injury resulting from oxidative and nitrosative stress, is considered to play a key role in the pathogenesis of diabetes complications by promoting numerous vascular dysfunctions. In this study, we examined the ability of metformin, which was reported to possess intrinsic vasculoprotective properties independently of its antihyperglycemic effects, to inhibit PARP activation induced by high glucose concentrations in bovine aortic endothelial cells; and we investigated the potential mechanisms involved in this inhibition. The PARP activity was measured by cellular enzyme linked immuno-specific assay (CELISA) method; cell poly(ribosyl)ated protein polymer accumulation was evaluated by immunofluorescence. Peroxynitrite anion productions were determined using dihydrorhodamine 123 fluoroprobe; and expression of p47phox subunit of nicotinamide adenine dinucleotide phosphate (NAD(P)H) oxidase was analyzed by Western blot in the absence and presence of protein kinase C and NAD(P)H oxidase inhibitors (calphostin and diphenyleneiodonium chloride, respectively). Our data showed that a therapeutically relevant concentration of metformin (5.10(-5) mol/L) was able to abolish PARP activation, to reduce poly(ribosyl)ated protein polymer accumulation, to decrease intracellular peroxynitrite anion level, and to reverse the overexpression of p47phox in bovine aortic endothelial cells stimulated by 25 mmol/L glucose in a similar manner to that of calphostin or diphenyleneiodonium chloride. Taken together, these results suggest that metformin could inhibit glucose-induced PARP activation through blockade of a protein kinase C-dependent NAD(P)H oxidase activation pathway. We propose that some of the beneficial effects of metformin on vascular endothelial cell functions in diabetes may be related to its inhibitory effect on PARP overactivation and its deleterious consequences. PMID- 19303975 TI - Red blood cell membrane alpha-linolenic acid and the risk of sudden cardiac arrest. AB - Higher levels of long-chain n-3 polyunsaturated fatty acids in red blood cell membranes are associated with lower risk of sudden cardiac arrest. Whether membrane levels of alpha-linolenic acid, a medium-chain n-3 polyunsaturated fatty acid, show a similar association is unclear. We investigated the association of red blood cell membrane alpha-linolenic acid with sudden cardiac arrest risk in a population-based case-control study. Cases, aged 25 to 74 years, were out-of hospital sudden cardiac arrest patients attended by paramedics in Seattle, WA (n = 265). Controls, matched to cases by age, sex, and calendar year, were randomly identified from the community (n = 415). All participants were free of prior clinically diagnosed heart disease. Blood was obtained at the time of cardiac arrest (cases) or at the time of an interview (controls). Higher membrane alpha linolenic acid was associated with a higher risk of sudden cardiac arrest: after adjustment for matching factors and smoking, diabetes, hypertension, education, physical activity, weight, height, and total fat intake, the odds ratios corresponding to increasing quartiles of alpha-linolenic acid were 1.7 (95% confidence interval [CI], 1.0-3.0), 1.9 (95% CI, 1.1-3.3), and 2.5 (95% CI, 1.3 4.8) compared with the lowest quartile. The association was independent of red blood cell levels of long-chain n-3 fatty acids, trans-fatty acids, and linoleic acid. Higher membrane levels of alpha-linolenic acid are associated with higher risk of sudden cardiac arrest. PMID- 19303976 TI - The Pro12Ala polymorphism of the PPARgamma2 gene is associated with hepatic glucose uptake during hyperinsulinemia in subjects with type 2 diabetes mellitus. AB - The Ala12 allele of the peroxisome proliferator-activated receptor gamma gene (PPARG2) has been associated with reduced risk of type 2 diabetes mellitus (T2DM) and increased whole-body and skeletal muscle insulin sensitivity in nondiabetic subjects. The effect of the Pro12Ala polymorphism on tissue specific insulin sensitivity in subjects with T2DM has not been previously investigated. We studied the effect of the Pro12Ala polymorphism on the rates of whole-body, skeletal muscle, and subcutaneous adipose tissue glucose uptake (GU) in T2DM subjects, and the rates of hepatic GU in nondiabetic and T2DM subjects during hyperinsulinemia. Our study included 105 T2DM subjects whose whole-body, skeletal muscle, subcutaneous adipose tissue, and hepatic GUs were measured using (18)F fluorodeoxyglucose and positron emission tomography during the hyperinsulinemic euglycemic clamp. Hepatic GU was also measured in 68 nondiabetic subjects. In obese (body mass index >or=27 kg/m(2)) subjects with T2DM, the rate of hepatic GU was 28% lower in subjects with the Pro12Pro genotype than in carriers of the Ala12 allele (P = .001); and a similar trend was observed in nondiabetic obese subjects (P = .137). No effect of the Pro12Ala polymorphism on the rates of whole body, skeletal muscle, or subcutaneous adipose tissue GU was observed in T2DM subjects. We conclude that the Ala12 allele of PPARG2 is associated with higher hepatic GU in obese subjects with T2DM. PMID- 19303977 TI - Serum levels of angiopoietin-related growth factor in diabetes mellitus and chronic hemodialysis. AB - Angiopoietin-related growth factor (AGF) was recently introduced as a novel liver derived protein that antagonizes obesity and insulin resistance. In the current study, we investigated circulating AGF levels in relation to renal function and type 2 diabetes mellitus (T2DM). Angiopoietin-related growth factor was determined by enzyme-linked immunosorbent assay in subjects with a glomerular filtration rate greater than 50 mL/min (n = 60, 30 diabetic and 30 nondiabetic) and in patients on chronic hemodialysis (CD; n = 60, 32 diabetic and 28 nondiabetic). Furthermore, AGF was correlated to clinical and biochemical measures of renal function, glucose and lipid metabolism, as well as inflammation. Median serum AGF levels were significantly lower in CD patients (125.9 +/- 96.3 microg/L) as compared with subjects with a glomerular filtration rate greater than 50 mL/min (164.0 +/- 95.4 microg/L) (P < .05). Furthermore, AGF serum levels were significantly increased in diabetic patients (161.7 +/- 114.2 microg/L) as compared with nondiabetic subjects (123.0 +/- 88.2 microg/L) (P < .01). Moreover, CD negatively and T2DM positively predicted AGF concentrations in multiple regression analysis. In addition, fasting serum glucose was independently and positively correlated with circulating AGF in all patients and controls. Our results suggest that renal dysfunction is negatively and T2DM is positively associated with AGF serum levels. Further studies are needed to better elucidate the physiologic significance of circulating AGF in human disease. PMID- 19303978 TI - Cold exposure increases adiponectin levels in men. AB - Sympathetic nerve activation is recognized at the adipose tissue level during cold exposure. Adiponectin is a key protein produced by adipose tissue, but its acute modulation remains unknown in humans exposed to cold. The aim of this study were (1) to examine the acute effects of cold exposure on circulating adiponectin and (2) to determine whether the changes are modulated by (a) an acute glucose ingestion as well as (b) a short-term modulation in carbohydrate (CHO) availability. Using a random crossover design, 6 healthy men were exposed to cold for 120 minutes with ingestion of beverages containing low (Control, 0.04 g/min) or high (High, 0.8 g/min) amounts of glucose during the course of the experiment (study 1). In study 2, 6 healthy men were exposed twice to cold for 120 minutes after equicaloric low-CHO diet and exercise and high-CHO diet without exercise. Plasma adiponectin concentrations were quantified before and during cold exposure. In study 1, adiponectin levels did not change during High, whereas a 20% rise was observed during Control (condition x time interaction, P = .06). In study 2, adiponectin levels increased by approximately 70% during cold exposure after both low- and high-CHO diets (effect of time, P < .05). A 120-minute period of cold exposure is accompanied by a significant increase in adiponectin levels in young healthy men. The rise in adiponectin levels observed during shivering is inhibited with glucose ingestion but not after diets varying in CHO content. PMID- 19303979 TI - Overexpression of apolipoprotein A5 in mice is not protective against body weight gain and aberrant glucose homeostasis. AB - Apolipoprotein A5 (APOA5) is expressed primarily in the liver and modulates plasma triglyceride levels in mice and humans. Mice overexpressing APOA5 exhibit reduced plasma triglyceride levels. Because there is a tight association between plasma triglyceride concentration and traits of the metabolic syndrome, we used transgenic mice overexpressing human APOA5 to test the concept that these mice would be protected from diet-induced obesity and insulin resistance. Male and female transgenic and wild-type mice on the FVB/N genetic background were fed standard rodent chow or a diet rich in fat and sucrose for 18 weeks, during which time clinical phenotypes associated with obesity and glucose homeostasis were measured. We found that APOA5 transgenic (A5tg) mice were resistant to diet induced changes in plasma triglyceride but not total cholesterol levels. Body weights were similar between the genotypes for females and males, although male A5tg mice showed a modest but significant increase in the relative size of inguinal fat pads. Although male A5tg mice showed a significantly increased ratio of plasma glucose to insulin, profiles of glucose clearance as evaluated after injections of glucose or insulin failed to reveal any differences between genotypes. Overall, our data showed that there was no advantage to responses to diet-induced obesity with chronic reduction of plasma triglyceride levels as mediated by overexpression of APOA5. PMID- 19303980 TI - Adenoviral expression of human lecithin-cholesterol acyltransferase in nonhuman primates leads to an antiatherogenic lipoprotein phenotype by increasing high density lipoprotein and lowering low-density lipoprotein. AB - Lecithin-cholesterol acyltransferase (LCAT), a key enzyme in high-density lipoprotein (HDL) metabolism, has been proposed to have atheroprotective properties by promoting reverse cholesterol transport. Overexpression of LCAT in various animal models, however, has led to conflicting results on its overall effect on lipoproteins and atherosclerosis. In this study, the effect of overexpression of LCAT in nonhuman primates on lipoprotein metabolism is examined. Human LCAT was expressed with adenovirus in squirrel monkeys (n = 8), resulting on day 4 in a 22-fold increase of LCAT activity (257 +/- 23 vs 5618 +/- 799 nmol mL(-1) h(-1), P < .0001). At its peak, LCAT was found to nearly double the level of HDL cholesterol from baseline (113 +/- 7 vs 260 +/- 24 mg/dL, P < .01). High-density lipoprotein formed after treatment with the adenovirus was larger in size, as assessed by fast protein liquid chromatography (FPLC) analysis. By kinetic studies, it was determined that there was a decrease in apolipoprotein (Apo) A-I resident time (0.373 +/- 0.027 vs 0.685 +/- 0.045 d(-1), P < .0001) and almost a doubling in the ApoA-I synthetic rate (22 +/- 2 vs 41 +/- 3 mg kg(-1) d(-1), P < .0001), but no overall change in ApoA-I levels. In addition, increased expression of LCAT was associated with a 37% reduction of ApoB levels (12 +/- 1 vs 19 +/- 1 mg/dL, P < .05) due to increased low-density lipoprotein catabolism (fractional catabolic rate = 1.7 +/- 0.1 d(-1) in controls vs 4.2 +/- 0.3 d(-1) in LCAT-treated group, P < .05). In summary, overexpression of LCAT in nonhuman primates leads to an antiatherogenic lipoprotein profile by increasing HDL cholesterol and lowering ApoB, thus making LCAT a potential drug target for reducing atherosclerosis. PMID- 19303981 TI - Elevated coagulation and inflammatory markers in adolescents with a history of premature adrenarche. AB - Females with a history of premature adrenarche are at high risk of developing polycystic ovary syndrome (PCOS) and features of the metabolic syndrome later in life. Coagulation disorders, subclinical inflammation, and oxidative stress have been reported in patients with PCOS and metabolic syndrome. These factors were studied in a group of adolescents with a history of premature adrenarche. This is a cross-sectional study that determined the biochemical-hormonal profile and indices of inflammation, coagulation, and oxidative stress in 45 adolescent girls with a history of premature adrenarche and 19 age- and body mass index-matched controls. Girls with premature adrenarche had hyperandrogenism and higher indices of insulin resistance than controls. They also had significantly higher C reactive protein (0.76 +/- 0.65 vs 0.41 +/- 0.31 mg/L, P = .0001) and plasminogen activator inhibitor 1 (37.6 +/- 24.7 vs 24.47 +/- 4.6 ng/mL, P = .034), and lower tissue plasminogen activator values in comparison with controls (3.5 +/- 1.5 vs 5.2 +/- 2.12 ng/mL, P = .0019). Both C-reactive protein(r = 0.545, P = .0001) and plasminogen activator inhibitor 1 (r = 0.36, P = .04) were positively correlated with oxidative stress, whereas tissue plasminogen activator was positively correlated (r = 0.37, P = .02) with total antioxidant status. None of these factors was correlated with androgens or indices of insulin resistance. Adolescent girls with a history of premature adrenarche display metabolic deviations usually encountered in subjects with PCOS and metabolic syndrome, such as subclinical inflammation and fibrinolytic abnormalities. PMID- 19303982 TI - Suppressing the suppressor: Role of immunosuppressive regulatory T cells in cancer surgery. PMID- 19303983 TI - Effects of a nonsurgical hospitalist service on trauma patient outcomes. AB - BACKGROUND: The American College of Surgeons criteria for Level I trauma centers calls for >90% of trauma patients to be admitted directly by a trauma surgeon or surgical subspecialist; however, the efficiency of the trauma system may be increased if patients presenting with comorbid conditions and minor injuries are treated by a hospitalist team (nonsurgical Trauma MEDical [TMED] service). We hypothesized outcomes would be equivalent for patients treated under TMED versus a surgical service. METHODS: This retrospective review compared mortality, hospital length of stay (LOS), Emergency Department (ED) LOS, placement to rehabilitation facilities, and complication rates for patients who could have been treated by TMED as identified by an algorithm. The study population for 2003 (pre-TMED) was compared with the study population for 2006 (post-TMED). Univariate analyses and multivariate logistic and linear regression were used to identify outcomes that were different for patients treated in 2003 versus 2006. Sensitivity, specificity, and percent kappa agreement were calculated for patients who were treated by the TMED team in 2006 versus patients in 2006 who were identified using the algorithm. RESULTS: The algorithm had reasonable sensitivity (78%) and specificity (90%); the kappa agreement was excellent (0.88). No differences were found in mortality (P = .31), rate of complications (P = .08), ED LOS (P = .77), or placement to rehabilitation facilities (P = .29) for patients identified in 2003 versus 2006. Hospital LOS was increased in 2006 (3.7 vs 4.1 days; P = .02). CONCLUSION: These data support admission of trauma patients with nonsevere, single-system injuries to a nonsurgical hospitalist service. We hypothesize that overall system efficiency may be improved by applying this alternative model in other trauma centers. PMID- 19303984 TI - Risk factors for chemotherapy-associated liver injuries: A multivariate analysis of a group of 146 patients with colorectal metastases. AB - BACKGROUND: Chemotherapy for colorectal liver metastases (CLM) may be associated with pathologic changes to the liver parenchyma, possibly increasing the risk of surgery. The aim of this study was to determine the risk factors for chemotherapy associated liver injuries (CALI). METHODS: From 1998 to 2006, 146 patients underwent a liver resection for CLM within 3 months of preoperative chemotherapy. CALI were defined as follows: steatosis > or =30%, or steatohepatitis, or moderate or severe sinusoidal lesions. Univariate and multivariate analysis were used to examine the relationship between CALI and 36 other preoperative factors. RESULTS: Pathologic examination showed CALI in 74/146 patients (51%), including sinusoidal lesions (n = 50), steatosis > or = 30% (n = 32), and steatohepatitis (n = 15). Twenty-three patients had overlapping CALI. In multivariate analysis, BMI > 27 (P = .002), hyperglycemia > 7 mmol/l (P = .006), and an interval between chemotherapy and surgery < 4 weeks (P = .011) were independent risk factors for CALI, whereas aspirin intake was associated with reduced risk of CALI (P = .002). In multivariate analysis, synchronous metastases (P = .015), unresectable liver metastases at diagnosis (P = .034), GGT plasma level > 1.5N (P = .028), and oxaliplatin-based chemotherapy (P < .01) were risk factors for sinusoidal lesions. Aspirin intake was associated with reduced risk for sinusoidal lesions (P = .03). In multivariate analysis, BMI > 27 (P = .026) was the only risk factor for steatosis or steatohepatitis. CONCLUSION: In patients with CLM, there are few preoperative risk factors for CALI. In patients treated by oxaliplatin-based chemotherapy, aspirin intake appears to be associated with a reduced risk of sinusoidal lesions and should be tested in a randomized phase II study. PMID- 19303985 TI - Erythropoietin reduces necrosis in critically ischemic myocutaneous tissue by protecting nutritive perfusion in a dose-dependent manner. AB - BACKGROUND: Erythropoietin (Epo), the primary regulator of erythropoiesis, has recently been shown to exert antiinflammatory and antiapoptotic properties in neuronal and myocardial tissue. We herein studied whether Epo pretreatment can reduce cell death and ischemic necrosis in a chronic in vivo model. METHODS: C57BL/6 mice were treated daily for 3 consecutive days with either 500 IU EPO/kg body weight (bw) (group Epo 500, n = 8) or 5000 IU EPO/kg bw (group Epo 5000, n = 8) administered intraperitoneally 24 hours before surgery. Thereafter, a random pattern myocutaneous flap subjected to acute persistent ischemia was elevated and fixed into a dorsal skinfold chamber. Flap elevation in animals receiving the water-soluble vitamin E analog Trolox (6-hydroxy-2, 5, 7, 8-tetramethylchroman-2 carboxylic acid) served as a nonspecific antiinflammatory agent control group (Tro); untreated control animals (Con) received saline only. Capillary perfusion, leukocyte-endothelial cell interaction, apoptotic cell death, and tissue necrosis were determined over a 10-day observation period using intravital multifluorescence microscopy. RESULTS: Epo 5000 (44 +/- 26 cm/cm(2)) but, more noticeably, Epo 500 (116 +/- 32 cm/cm(2)) improved capillary perfusion compared with the two control groups, particularly the Con group (9 +/- 7 cm/cm(2); P < .05). The ischemia-associated leukocytic inflammation was found drastically attenuated in both Epo-pretreatment groups. Epo 500 further decreased apoptotic cell death and was effective in significantly reducing tissue necrosis (16% +/- 4% vs Tro: 48% +/- 7% and Con: 52% +/- 4%; P < .001). No angiogenic blood vessel formation could be observed in either of the Epo groups. Of interest, Epo 5000 but not Epo 500-increased systemic hematocrit. CONCLUSION: Despite the lack of neovascularization, Epo pretreatment was capable of reducing ischemic tissue necrosis by protecting capillary perfusion, ie, nutrition of the tissue. Low-dose pretreatment was more effective, a result that was most likely due to the better perfusion conditions without an increase of the hematocrit values. Thus, low-dose Epo pretreatment might represent a promising strategy to protect critically perfused ischemic tissue. PMID- 19303986 TI - L-selectin shedding in sepsis limits leukocyte mediated microvascular injury at remote sites. AB - BACKGROUND: Increased soluble L-selectin levels have been shown to attenuate local inflammation-mediated microvascular leakage, and failure to generate high levels has been associated with increased risk of acute respiratory distress syndrome in septic patients. We hypothesized that failure to shed L-selectin in systemic inflammation would result in increased local inflammation-induced leukocyte adherence and microvascular leakage. METHODS: Using intraperitoneal lipopolysaccharide (LPS) or control bicarbonate buffered saline (BBS) and intrascrotal TNFalpha or BBS, mice were randomized to systemic inflammation (LPSip + BBSis), local inflammation (BBSip + TNFis), both (LPSip + TNFis), or control (BBSip+BBSis). Furthermore, mice received intraperitoneal L-selectin Sheddase inhibitor (Ro31-9790) or control vector. With intravital microscopy on cremaster muscle, we measured leukocyte-endothelial cell interactions and microvascular leakage (permeability index). Surface L-selectin was measured by flow cytometry (MCF). RESULTS: Without Ro31-9790, systemic inflammation attenuated increases induced by local inflammation in leukocyte adherence and vascular leakage. Ro31-9790 significantly increased adherence and leakage in systemic and systemic + local inflammation. L-selectin was shed progressively by increasing degrees of inflammation. Ro31-9790 limited this shedding of L selectin. CONCLUSION: In systemic inflammation, L-selectin shedding is required to limit local inflammation-mediated leukocyte adherence and microvascular leakage. Failure to shed L-selectin may increase leukocyte-mediated end-organ injury in septic patients. PMID- 19303987 TI - Thymectomy is more effective than conservative treatment for myasthenia gravis regarding outcome and clinical improvement. AB - BACKGROUND: Myasthenia gravis (MG) is an autoimmune disease with a tremendous impact on the quality of life. Controversies over which patients should be operated on because they may benefit most from thymectomy are still ongoing. The aim of this study was to report our long-term results of patients with MG with comparison of thymectomy and conservative treatment. METHODS: We report a series of 252 patients with MG. Survival data were generated. Patients were seen in the outpatient clinic, where a modified Osserman score and quality of life score were evaluated at the end of the follow-up period for all surviving patients. RESULTS: A total of 172 patients with MG were followed after thymectomy or with conservative treatment for a median time of 9.8 years. Patients who underwent thymectomy had significantly greater rates of remission and improvement compared with conservative treatment. Furthermore, they had a significantly greater survival. CONCLUSION: Currently, different effective modalities of treatment are available in patients with MG. In our long-term follow-up, thymectomy was superior to conservative treatment regarding overall survival, clinical improvement, and remission rate. Therefore, thymectomy should be considered strongly for all patients with generalized MG. PMID- 19303988 TI - Major hepatic resection for hepatocellular carcinoma with or without portal vein embolization: Perioperative outcome and survival. AB - BACKGROUND: Preoperative portal vein embolization (PVE) is performed to minimize perioperative risks of major hepatic resection for hepatocellular carcinoma (HCC), but its effects on tumor growth are ill defined. Perioperative outcome and survival after major hepatic resection for HCC, with and without PVE, were investigated. METHODS: Patients that underwent major hepatic resection (> or =3 segments) for HCC between January 1998 and May 2007 were analyzed retrospectively. Preoperative PVE was performed when the remnant liver volume was predicted to be insufficient. RESULTS: A total of 54 patients underwent major hepatic resection for HCC: 21 patients with PVE before resection (PVE group) and 33 patients without PVE (non-PVE group). PVE and non-PVE groups had similar rates of fibrosis or cirrhosis, hepatitis C virus, hepatitis B virus, American Joint Committee on Cancer stage, preoperative transarterial chemoembolization, overall postoperative complications, and positive margin (P = nonsignificant for all rates). There were no perioperative deaths in the PVE group and 6 (18%) deaths in the non-PVE group (P = .038). Median follow-up was 21 months. Excluding perioperative deaths, overall survival rates at 1, 3, and 5 years were 94%, 82%, and 72%, respectively, in the PVE group and 93%, 63%, and 54%, respectively, in the non-PVE group (P = .35). Similarly, disease-free survival (DFS) rates were not significantly different between the groups, with 1-, 3-, and 5-year DFS rates of 84%, 56%, and 56%, respectively, in the PVE group and 66%, 49%, and 49%, respectively, in the non-PVE group (P = .38). CONCLUSION: PVE before major hepatic resection for HCC is associated with improved perioperative outcome. Excluding perioperative mortality, overall survival and DFS rates were similar between patients with and without preoperative PVE. PMID- 19303989 TI - Recanalized umbilical vein conduit for meso-Rex bypass in extrahepatic portal vein obstruction. AB - BACKGROUND: Meso-Rex bypass is used to treat patients with clinically important extrahepatic portal vein obstruction (EHPVO). Usually, an autologous left internal jugular vein graft is used to bypass the portal blood circulation from the superior mesenteric vein to the left portal vein. Other vascular conduits have included the autogenous saphenous vein, splenic vein, right gastroepiploic vein, and inferior mesenteric vein. METHODS: A total of 20 umbilical veins with attached livers were harvested from 20 deceased liver donors. Umbilical veins were dilated mechanically and checked for patency and communication with the left portal vein. Vein length and diameter after dilatation were recorded. Cross sections of 15 recanalized umbilical veins were processed by routine histologic examination and stained with hematoxylin and eosin, as well as processed by immunohistochemistry for CD31 and factor VIII antigens. Subsequently, 3 children with EHPVO underwent this modified meso-Rex bypass using the umbilical vein as a vascular conduit. RESULTS: The mean length of harvested umbilical veins was 15 cm (range, 7-21); the mean length of recanalized and usable umbilical veins was 10 cm (range, 5-15). Recanalization was successful in 16 (80%) of the 20 donor umbilical veins. The mean diameter of the umbilical veins after serial dilatation and recanalization was 1.2 cm (range, 1-2). In 11 (73%) of the 15 recanalized vein specimens, the lumen was lined by endothelial cells. In 2 children, the vascular conduit was constructed entirely with native umbilical vein. In the remaining child, 3 cm of umbilical vein was preserved and anastomosed to a mobilized inferior mesenteric vein due to inadequate length. All 3 children had patent bypass and resolution of clinical manifestations of portal hypertension at a mean follow-up of 21 months. CONCLUSION: Meso-Rex bypass may prove to be a definitive treatment for patients with EHPVO. The use of native umbilical vein as a vein conduit achieved decompression of the splanchnic venous system and should be considered a natural alternative to other interposition vein grafts. PMID- 19303990 TI - Therapeutic value of lymph node dissection during hepatectomy in patients with intrahepatic cholangiocellular carcinoma with negative lymph node involvement. AB - BACKGROUND: Routine and radical lymph node dissection is a clinical concern for improving the surgical outcome in patients with intrahepatic cholangiocarcinoma (ICC). The therapeutic value of the procedure during hepatectomy has, however, not been evaluated. METHODS: Between January 1990 and December 2004, 104 patients with ICC undergoing macroscopic curative resections were investigated retrospectively with special reference to lymph node status. The role of lymph node dissection was evaluated according to macroscopic type: mass-forming (MF) type (n = 68) and MF plus periductal infiltration (PI) type (n = 36) of ICC. RESULTS: Lymph node involvement and intrahepatic metastases were an independent, unfavorable prognostic factor in the MF type of ICC. Negative lymph node involvement provided a favorable survival rate in the 41 patients without intrahepatic metastases (P < .0001). Among the 29 patients without lymph node involvement and intrahepatic metastases, there was no difference according to the use of lymph node dissection (P = .8071). Also, no difference was seen with lymph node involvement in the 24 patients with the MF plus PI type of ICC who had no intrahepatic metastases (P = .6620). CONCLUSION: For purpose of diagnostic staging and exclusion of positive regional lymph nodes, lymph node dissections might be useful in patients with the MF type and the MF plus PI type of ICC; however, routine use of lymph node dissection in patients with the MF type of ICC is not recommended, because no difference in survival was observed in the patients with negative lymph node metastases, irrespective of the use of lymph node dissection. PMID- 19303991 TI - Portal or superior mesenteric vein resection for pancreatic head adenocarcinoma: prognostic value of the length of venous resection. AB - BACKGROUND: We investigated retrospectively the operative outcomes of portal vein resection (PVR) for portal vein (PV) and/or superior mesenteric vein (SMV) involvement and clarified the validity of PVR. METHODS: Between 1993 and 2006, 84 patients with invasive pancreatic head adenocarcinoma were resected by pancreatoduodenectomy with macroscopically curative resection. Forty-two patients underwent PVR by means of segmental resection with end-to-end anastomosis in 27 patients and autologous vein graft using an external iliac vein in 15 patients because of macroscopic venous involvement. Venous involvement was classified macroscopically as unilateral involvement (< or =180 degrees ; n = 27) or circumferential involvement (n = 15) and as short (the length of PVR < 3 cm; n = 15) or long (> or =3 cm; n = 27). Histopathologic parameters and survival were analyzed to confirm prognostic factors. RESULTS: Morbidity and mortality were not different based on PVR status. Median and 5-year survivals were 26 months and 32%, respectively, when there was no PVR (n = 42) and 12 months and 17% when there was PVR (n = 42); these values of median and 5-year survivals differed (P < .04 each) between the groups without and with PVR. Limiting the analysis to R0 (histologically curative) resections, median and 5-year survivals were 26 months and 34% when there was no PVR (n = 39) and 20 months and 23% when there was PVR (n = 32); these survivals were not significantly different between groups. In patients with PVR, there were no statistical differences in survival between those resected with or without a venous allograft and those with unilateral or circumferential involvement; however, short PVR showed better 5-year survival than long PVR (39% vs 4%; P = .017) despite similar positive rates of histologic venous invasion. CONCLUSION: PVR has comparable survival compared with no PVR only in patients undergoing an R0 resection. The short PV/SMV invasion that requires PVR <3 cm in length can result in respectable survival rates. PMID- 19303992 TI - Revascularization to an isolated ("blind") popliteal artery segment: a viable procedure for critical limb ischemia. AB - BACKGROUND: The purpose of this study was to analyze our experience of bypass procedures to an isolated ("blind") popliteal artery segment (IPAS) to revascularize the perigeniculate arteries in patients with critical limb ischemia (CLI), to establish whether such revascularizations could yield acceptable results in terms of patency and limb salvage (LS) rates. METHODS: Over a decade, 347 above-knee arterial revascularizations were performed in 293 patients and in 51 (14.7%) of these the outflow vessels were the perigeniculate arteries arising from an IPAS, through a reversed saphenous vein or spliced veins (n = 30, 58.8%; group I) or polytetrafluoroethylene (n = 21, 41.2%; group II) prosthetic grafts. Patency, LS, and survival rates were assessed using Kaplan-Meier life-table analysis. A complete follow-up (range, 0.1-10.4 years; mean, 5.6 years) was obtained in 49 patients. RESULTS: The IPAS was chosen as the last resort in 39 patients (76.5%) because no other infrapopliteal artery was identified as being available at angiography; in 12 patients (23.5%) it was chosen because of an invasive foot infection or ischemic necrosis overlying the dorsalis pedis or the posterior tibial arteries. The study series was mainly male, with significantly more younger patients in group I (72 +/- 1 years vs 74 +/- 5 years, P = .037). Group I had a statistically higher incidence of diabetes mellitus (76.6% vs 47.6%, P = .033), insulin dependence (56.7% vs 28.6%, P = .047) and history of smoking (80% vs 47.6%, P = .016) than group II. None of the patients died in the perioperative period. There were 3 early graft failures (2 in group I), prompting 3 major amputations. Kaplan-Meier analysis identified 5-year patency and LS rates of 51.4 +/- 9.6% and 90 +/- 4.3%, respectively, in the series as a whole, and the 2 groups had comparable 5-year patency, LS and survival rates. CONCLUSION: Revascularizations to an IPAS can be performed with acceptable results in terms of patency and LS rates, even when there is no infrapopliteal runoff vessel. Finding perigeniculate arteries arising from an IPAS with no tibio-peroneal vessel reconstitution at arteriography does not justify a pessimistic attitude to the performance of such revascularizations for LS. PMID- 19303993 TI - Bacterial population moves toward a colon-like community in the pouch after total proctocolectomy. AB - BACKGROUND: Colonic transformation is defined by phenotypic alterations in the ileum after total proctocolectomy. Changes in microbiota of the ileal pouch and the roles of these microbes in colonic transformation, however, have not been addressed. METHODS: A total of 151 stool samples were collected from patients with ulcerative colitis patients and an ileostomy, those with an ileal pouch, and healthy control volunteers. Bacterial DNA was extracted from stool, and the diversity of complex bacteria was assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis, a novel DNA-based approach that enables us to investigate the presence of nonculturable bacteria. To determine whether ileal pouch bacterial communities shift to a more colon-like distribution, the relative abundance of terminal restriction fragments that could be classified as "colonic," "ileal," or "common" was investigated. RESULTS: Cluster analysis demonstrated that most of the ileostomy samples were categorized into Cluster I or II and that less than 10% of ileostomy samples were classified into Cluster IV. In contrast, more than 90% of control samples were grouped in Cluster IV. In further analyses, the median lifetimes of pouches in Clusters I, II, III, and IV were significantly different at 11, 56, 265, and 310 days, respectively. T-RFLP patterns of the ileal pouch were characterized by a time-dependent decrease in "ileal" and increase in a part of "colonic" fragments, which represented mainly nonculturable bacteria such as the Clostridium coccoides group. CONCLUSION: T RFLP analysis demonstrated that a time-dependent shift to a "colon-like" bacterial community, including nonculturable bacteria, in the ileal pouch after total proctocolectomy. PMID- 19303994 TI - Torsion and infarction of accessory liver lobe in young man. PMID- 19303995 TI - Persistent sciatic artery. PMID- 19303996 TI - Benefit of laparoscopy for rectal resection in patients operated simultaneously for synchronous liver metastases: preliminary experience. PMID- 19303997 TI - Advantages of the first-step totally laparoscopic approach in 2-staged hepatectomy for colorectal synchronous liver metastasis. PMID- 19303998 TI - Dear Gordon: can we discuss the G20? PMID- 19303999 TI - Medical education and professionalism. PMID- 19304000 TI - Early colorectal cancer screening for African Americans. PMID- 19304001 TI - Engaging the health community in global economic reform. PMID- 19304002 TI - The French salt industry in court. PMID- 19304003 TI - Protocols, probity, and publication. PMID- 19304004 TI - Geoff Foster: paediatrician remaining hopeful in Zimbabwe. PMID- 19304005 TI - Migraine and telcagepant [corrected]. PMID- 19304007 TI - The Lancet and advertorials. PMID- 19304009 TI - Mechanisms of thrombogenesis in atrial fibrillation. PMID- 19304011 TI - Mechanisms of thrombogenesis in atrial fibrillation. PMID- 19304014 TI - Atypical antipsychotic drugs have their merits. PMID- 19304015 TI - Measles elimination: lack of progress in the Western Pacific Region. PMID- 19304016 TI - Monoclonal antibodies for cancer immunotherapy. AB - Monoclonal antibodies are effective treatments for many malignant diseases. However, the ability of antibodies to initiate tumour-antigen-specific immune responses has received less attention than have other mechanisms of antibody action. We describe the rationale and evidence for the development of antibodies that can stimulate host tumour-antigen-specific immune responses. Such responses can be induced through the induction of antibody-dependent cellular cytotoxicity, promotion of antibody-targeted cross-presentation of tumour antigens, or by triggering of the idiotypic network. Future treatment modifications or combinations might be able to prolong, amplify, and shape these immune responses to increase the clinical benefits of antibody therapy for human cancer. PMID- 19304017 TI - Opening up public health: a strategy of information and communication technology to support population health. PMID- 19304018 TI - Loss of sight and sound. Could it be the hip? PMID- 19304025 TI - "Sully" Sullenberger and the miracle on the Hudson: a lesson in heroism for oral and maxillofacial surgeons. PMID- 19304026 TI - Outcomes of placing short dental implants in the posterior mandible: a retrospective study of 124 cases. AB - PURPOSE: The purpose of this retrospective study was to determine the overall success of short dental implants (8 mm in length) placed in the partially or completely edentulous posterior mandible restored with fixed and removable prostheses. PATIENTS AND METHODS: A total of 124 patients had 335 8-mm-long implants placed from May 2005 until June 2007. Of the 124 total patients, 35 were men and 89 were women, with a median age of 56 years and an age range of 18 to 80 years at the time of implant surgery. There were 112 patients who were partially edentulous and 12 who were completely edentulous. Of the patients, 32 had a single implant placed whereas the other 92 had multiple implants placed. One patient had the implants immediately provisionally loaded. All of the implants were restored by use of fixed prostheses. Of these fixed prostheses, 245 were splinted together whereas 75 were restored individually. RESULTS: A total of 335 short dental implants were placed in 124 patients. Of the 335 implants placed, 331 integrated successfully. In the 2 cases that failed, the sites were grafted with porous hydroxyapatite and platelet-rich plasma. The implants were replaced at 5 months after the initial failure in the first patient and at 7 months in the second patient. These replacement implants integrated and have been restored and in function for more than 16 months. There was 1 fracture of an implant with a restoration. The implant had been restored with an individually fabricated fixed restoration, with the fracture occurring at the head of the implant, requiring removal. The implant and restoration had been in function for 10 months before fracture.There were no other fractures of implants or restorative hardware noted in this study. The survival rate for 8-mm implants placed in the mandible was 99% from stage I surgery to a functional prosthesis for up to 2 years. CONCLUSIONS: Placement of short dental implants is a predictable treatment method for patients with decreased posterior mandibular bone height. PMID- 19304027 TI - Metastatic tumors to the oral and maxillofacial region: a retrospective study of 19 cases in West China and review of the Chinese and English literature. AB - PURPOSE: The aim of this article was to obtain an overview of metastatic tumors to the oral and maxillofacial (OMF) region, especially the differences in the constituent ratios of primary cancers between the United States and China. PATIENTS AND METHODS: Clinical findings of 19 cases encountered in West China Hospital of Stomatology were summarized and the English and Chinese literature were reviewed and analyzed. The main clinical features of OMF metastases were summarized, with an emphasis on primary cancers' constituents. RESULTS: The lung, breast, kidney, liver, and prostate were the top 5 common primary sites of cancer. However, there was a significant difference in the primary cancers' constituents between United States and China (P < .001). The breast, kidney, prostate cancers, and melanoma of skin were more frequent primary cancers in United States than in China, whereas that of the lung, thyroid, liver, esophagus, and the stomach were more common in China than in United States. The proportions of the OMF metastatic lesions originating in the lung, kidney, liver, thyroid, and esophagus in all OMF metastatic tumors were higher than the corresponding primary cancers' prevalent proportions. CONCLUSIONS: The frequency of developing OMF metastasis is not always consistent with primary cancers' prevalence, which suggests that different cancers have different potentiality to develop OMF metastasis. Cancers of the kidney, liver, lung, thyroid, and esophagus were more likely to spread to the OMF region. In general screening of primary cancer, it would be helpful to take into account the metastatic potentiality of different cancers and primary cancers' prevalence in different countries in the case of occult primary. PMID- 19304028 TI - Comparative study of the mechanical resistance of 2 separate plates and 2 overlaid plates used in the fixation of the mandibular condyle: an in vitro study. AB - PURPOSE: The objective of this study was to carry out a comparative evaluation of the mechanical resistance of 2 rigid internal fixation techniques for fractures of the mandibular condyle using miniplates. MATERIALS AND METHODS: Forty polyurethane resin replicas of human hemimandibles were used. The hemimandibles were sectioned to simulate a high subcondylar fracture and then stabilized with 2 fixing techniques using 2.0-mm system plates and screws. The fixation techniques were 2 separate 4-hole plates with 8 screws, and 2 overlaid 4-hole plates with 4 screws. Each system was submitted to load tests, with the application of the load in mediolateral and anteroposterior directions in an Instron 4411 universal assay machine (Instron, Norwood, MA). RESULTS: Load values and peak displacement were measured. Means and standard deviations were evaluated by analysis of variance (P < .05) and Tukey tests, in which it was verified that the antero-posterior peak load value was affected by the arrangement of the plates on the models, although no differences were observed between the groups for the mediolateral peak load. The arrangement of the plates did not have any influence on peak displacement. Similarly, the final value of the mediolateral load was not affected by the arrangement of the plates on the model. CONCLUSION: The experimental model with 2 separate plates was statistically superior to the model with 2 overlaid plates only in relation to anteroposterior peak load. Despite showing superiority in mediolateral peak load and peak displacement, there was no statistical difference between the groups for these parameters. PMID- 19304029 TI - Measurement of anterior loop length for the mandibular canal and diameter of the mandibular incisive canal to avoid nerve damage when installing endosseous implants in the interforaminal region: a second attempt introducing cone beam computed tomography. AB - PURPOSE: To measure and compare the anterior loop length (ALL) for the mandibular canal and the mandibular incisive canal diameter (ICD) at its origin in cadavers using anatomy and cone beam computed tomography (CBCT) to safely install endosseous implants in the most distal area of the interforaminal region. MATERIALS AND METHODS: The ALL and ICD were measured using CBCT in 4 cadavers, and using anatomy in 71 cadavers. RESULTS: The ranges and mean +/- SD for the anatomic measurements were: ALL, 0.0 to 9.0 mm and 1.9 +/- 1.7 mm; ICD, 1.0 to 6.6 mm and 2.8 +/- 1.0 mm. The average discrepancies between CBCT and anatomic measurements were 0.06 mm or less for both the ALL and the ICD, which were less than the resolution of CBCT. CONCLUSIONS: Because large variations in measurements were observed, both for ALL and ICD, no fixed distance mesially from the mental foramen should be considered safe. The ALL and the ICD can be estimated from the CBCT measurement. The preoperative CBCT measurement yields important information for each case. PMID- 19304030 TI - Accuracy of computer programs in predicting orthognathic surgery soft tissue response. AB - PURPOSE: The aim of the present systematic review is to investigate the accuracy of computer programs in predicting soft tissue response subsequent to skeletal changes after orthognathic surgery. MATERIALS AND METHODS: A systematic computerized database search was conducted using Ovid Medline, Ovid Medline in process and other nonindexed citations, all EBM reviews (including Cochrane DSR, ACP Journal, DARE, CCTR), Embase Experta Medica, Pascal, and Thomsen's ISI Web of Science databases. The terms orthognathic surgery, orthodontic treatment, computer prediction, computer simulation, growth prediction, accuracy, precision, forecast, and human studies were used for the searches. The reference lists of all the selected articles were also searched for any potential articles that might have been missed in the electronic search of the database, and additional information not available through the articles was obtained directly from the source (by contacting the author). RESULTS: Out of the 40 initially identified articles only 7 articles fulfilled the final selection criteria. All 7 studies in general showed accurate prediction outcomes (less than 2 mm) when compared with actual results in both horizontal and vertical directions. From all the available computer prediction programs Quick Ceph, DentoFacial Planner (DFP), and Computer Assisted Simulation System for Orthognathic Surgery (CASSOS) were the most frequently used. CONCLUSIONS: The most significant area of error in prediction through the available computer prediction programs was the lower lip area. Although the individual errors are almost always minimal (less than 2 mm), the composite addition of these minimal errors could have clinical implications. PMID- 19304031 TI - Surgical repositioning of the premaxilla with bone graft in 50 bilateral cleft lip and palate patients. AB - PURPOSE: The aim of this study was to evaluate a modified surgical technique for premaxilla repositioning with concomitant autogenous bone grafting in bilateral trans-foramen cleft lip and palate patients. PATIENTS AND METHODS: The study included 50 bilateral trans-foramen cleft lip and palate patients. Bone graft was harvested from the mandibular symphysis in 24 patients. Whenever more grafting was necessary, the iliac crest bone was used as the donor site (26 patients). The premaxilla was displaced by rupturing the bone and the palatine mucosa, and repositioned in a more adequate position using a surgical guide. The premaxilla and the grafts were fixed with miniplates and screws or screws only. The surgical guide was kept in place for 2 months, whereas the miniplates and screws were removed after 6 months, together with the complete bilateral lip and nose repair. Follow-up examinations were performed at 3, 6, and 12 months by means of periapical and occlusal radiographs, and by clinical examination. Thereafter, the patients were referred for completion of the orthodontic treatment. RESULTS: Overall, in 48 cases (96%) the treatment achieved total graft integration, with complete closure of the bucconasal and palatal fistulas, and premaxilla stability (either at first surgery or after reoperation). In the remaining 2 patients (4%), the treatment failed, due to necrosis of the premaxilla. CONCLUSIONS: The procedure is complex and involves risk. However, the patient's social inclusion, especially at the addressed age group, is the best benefit achieved. PMID- 19304032 TI - Result of maxillomandibular fixation using intraoral cortical bone screws for condylar fractures of the mandible. AB - PURPOSE: Treatment of mandibular condylar fractures is not standardized. The maxillomandibular cortical bone screw fixation technique carries many advantages. The aim of this work was to evaluate this technique for routine method. MATERIALS AND METHODS: Fifty patients treated by maxillomandibular fixation (MMF) by use of cortical bone screws from 2004 to 2006 were retrospectively analyzed. In our maxillofacial surgery unit in Bordeaux, France, our indication is to treat extra articulated fractures without severe displacement by MMF. RESULTS: The mean time required for MMF was 13 minutes, and fixation occurred after a mean of 16 days. Screw removal was performed after a mean of 26 days, and this required local anesthesia. Of the patients, 48 had good occlusion. Two patients had persistent lateral cross bites. Two patients had mandible deviation when they opened their mouths, and mouth opening was limited in one patient. Two patients had temporomandibular joint pain. CONCLUSIONS: MMF screws have more advantages and fewer disadvantages than arch bars when closed treatment has been selected as the treatment of choice. PMID- 19304033 TI - Assessment of difficulty in third molar surgery--a systematic review. AB - PURPOSE: The aim of this review was to identify the most important variables that determine surgical difficulty of impacted third molar extractions by their consistent showing in previous studies. MATERIALS AND METHODS: Electronic library search for current evidence in the world literature was conducted, and relevant articles were selected, scrutinized, and the findings were compared. RESULTS: Seven articles were most relevant, and the results of the comparison of the selected articles showed that demographic variable, age; operative variables: surgeon procedure type and number of teeth extracted; and ratiographic variable, depth angulation; and root morphology, are the most consistent determinants of difficulty. CONCLUSION: Current evidence is in support of 3 categoreis of variables strongly associated with surgical diffulty of impacted third molars. The most important variables in each of these categories have been identified. PMID- 19304034 TI - Influence of different types of guns, projectiles, and propellants on patterns of injury to the viscerocranium. AB - PURPOSE: Gunshot injuries to the oral and craniomaxillofacial region vary with the type of gun used. Computed tomography (CT) is the standard diagnostic tool for assessing tissue damage in patients with gunshot injuries. Cone-beam computed tomography (CBCT) is a new imaging technique that has recently become available for clinical diagnosis. The objective of this study was to characterize injury patterns with a focus on gun types and identify the imaging modality that is best suited to assessing injuries caused by different gun types. PATIENTS AND METHODS: We present the cases of 14 patients who sustained gunshot injuries to the viscerocranium during the past 10 years. The injuries were caused by 8 basic combinations of handguns or long guns with soft lead core projectiles, partial or full metal-jacketed bullets, and different propellants. Diagnosis was based on clinical and radiological findings (including CT and CBCT). RESULTS: We found a direct correlation between the gun/projectile combination on the one hand and the diameter of the wound track, tissue contamination, and tissue destruction on the other. Entrance and exit wounds are indicative of certain gun/projectile combinations. High-density projectiles cause severe artifacts in CT, unlike CBCT, making it difficult to evaluate anatomic structures in close proximity to the projectile. CONCLUSIONS: Every gun/projectile combination is associated with a typical pattern of injury. Even in the absence of the offending projectile, it is thus possible to narrow down the likely gun and/or projectile. In the diagnostic imaging of injuries caused by high-density projectiles, CBCT is more suitable than CT. PMID- 19304035 TI - Mechanical strength and stiffness of the biodegradable SonicWeld Rx osteofixation system. AB - PURPOSE: To determine the mechanical strength and stiffness of the new 2.1 mm biodegradable ultrasound-activated SonicWeld Rx (Gebruder Martin GmbH & Co, Tuttlingen, Germany) osteofixation system in comparison with the conventional 2.1 mm biodegradable Resorb X (Gebruder Martin GmbH & Co) osteofixation system. MATERIALS AND METHODS: Plates and screws were fixed to 2 polymethylmethacrylate blocks to simulate bone segments and were subjected to tensile, side bending, and torsion tests. During testing, force and displacement were recorded and graphically presented in force-displacement diagrams. For the tensile tests, the strength of the osteofixation system was measured. The stiffness was calculated for the tensile, side bending, and torsion tests. RESULTS: The tensile strength and stiffness as well as the side bending stiffness of the SonicWeld Rx system presented up to 11.5 times higher mean values than the conventional Resorb X system. The torsion stiffness of both systems presents similar mean values and standard deviations. CONCLUSIONS: The SonicWeld Rx system is an improvement in the search for a mechanically strong and stiff as well as a biodegradable osteofixation system. Future research should be done to find out whether the promising in vitro results can be transferred to the in situ clinical situation. PMID- 19304036 TI - A quantitative evaluation of inflammatory cells in human temporomandibular joint tissues from patients with and without implants. AB - PURPOSE: Previous studies made only qualitative assessments of immune cell responses to temporomandibular joint (TMJ) implant wear debris. The aim of this study was to perform a quantitative comparison of inflammatory cell types in TMJ tissues with implant wear debris, TMJ tissues with a history of disc pathology without implant debris, and TMJ tissues from normal control subjects. MATERIALS AND METHODS: TMJ tissues were collected from the following 3 groups of subjects: 1) individuals with failed TMJ implants (implant group, n = 10), 2) patients with TMJ disc pathology but no history of implant placement (nonimplant surgery group, n = 10), and normal cadaveric tissues with no history of surgery (control group, n = 10). Tissue sections (5 microm) from all subjects were stained with hematoxylin-eosin, after which cell counts were done for 2 types of inflammatory cells: multinucleated giant cells and lymphocytes. Mean inflammatory cell counts from the 3 groups were compared by use of a 1-way analysis of variance procedure and Bonferroni adjustment to maintain an overall type I error rate of .05. RESULTS: Implant group tissues contained significantly more inflammatory cells than tissues from the nonimplant surgery and control groups (P < .0001). Multinucleated giant cells were only present in implant group tissues. Although the high number of multinucleated giant cells present in the implant group obscured a total count of lymphocytes for that group, lymphocyte cell counts were still significantly greater (P < .005) in implant group tissues than in tissues from the other 2 groups. CONCLUSIONS: Our data provide quantitative confirmation that the presence of Proplast-Teflon implant (Vitek, Houston, TX) wear debris is associated with a significant increase in the number of local multinucleated giant cells and lymphocytes. PMID- 19304037 TI - The chronologic prevalence of temporomandibular joint disorders associated with bilateral intraoral vertical ramus osteotomy. AB - PURPOSE: The purpose of this study was to evaluate the long-term stability of the improvement of symptoms associated with temporomandibular joint (TMJ) disorders after intraoral vertical ramus osteotomy for the treatment of mandibular prognathism. MATERIALS AND METHODS: A total of 217 patients who had undergone bilateral intraoral vertical ramus osteotomy (BIVRO) from 1998 to 2005 were evaluated preoperatively and 1, 3, 6, 12, 18, and 24 months postoperatively regarding mouth opening, clicking, and pain of the TMJ. A retrospective study was conducted based on the results. RESULTS: Remarkable improvement of TMJ symptoms after BIVRO was reliable. Preoperative TMJ sounds disappeared after BIVRO in 94.3% of joints, and most of the joints that were sound free preoperatively remained without TMJ sounds postoperatively (98.2%). However, 19 joints exhibited recurrence, and transient TMJ sounds were observed in 35 joints. Preoperative TMJ pain had improved in 97.9% of joints at 1 month postoperatively, and TMJ pain was not observed in any joints at 18 months after surgery. Joints that were pain free before surgery remained without pain, although there were 20 joints that exhibited transient TMJ pain. The mean mouth opening was 50.0 mm before surgery, which decreased to 34.92 mm at 1 month postoperatively. This was followed by an increase to 44.44 mm to 48.75 mm at 6 months postoperatively, and thereafter mouth opening showed 94.72% to 97.5% recovery compared with the preoperative state. CONCLUSIONS: BIVRO can be used as a method of choice for relieving undesirable TMJ symptoms such as sound and pain, as well as for repositioning the condyle head to its physiologic position. Such favorable effects of BIVRO on the TMJ were not remarkably affected with time. PMID- 19304038 TI - Infection rate in mandibular angle fractures treated with a 2.0-mm 8-hole curved strut plate. AB - PURPOSE: The aim of this study was to determine the rate of postoperative infection and the efficacy of removing teeth in the line of mandibular angle fractures treated with 2.0-mm 8-hole titanium curved strut plates. Our understanding is that this method of repair is currently being used only in a few centers in the United States. MATERIALS AND METHODS: A retrospective review of mandibular angle fractures treated with a 2.0-mm 8-hole strut plate during a 4 year period. Postoperative antibiotics were given for 1 week. Follow-up appointments were 4 weeks or longer. A nonchewing diet was instructed for 6 weeks. Data for all selected patients include the information such as age, gender, etiology of injuries, medical history, concurrent injuries, nerve deficits, pre- and postoperative antibiotic administration, postop infection, a presence or absence of teeth in the line of fractures, and whether these teeth were removed. RESULTS: Four patients (4 of 49 or 8.2%) developed infections. Two of those patients had a tooth in the line of a fracture that was retained (2 of 14 or 14%). The third had a tooth in the line of a fracture that was extracted (1 of 18 or 5.6%). The fourth patient was 1 of the 17 patients who did not have teeth in the line of fracture and developed infection (1 of 17 or 5.9%). None of the patients developed failed hardware, malunion, nonunion, malocclusion, or iatrogenic nerve injury. CONCLUSIONS: The use of a 2.0-mm 8-hole strut plate is associated with a low infection rate (8.2%). The infection rate for those mandibular angle fractures with teeth in the line of fracture retained was 14% compared with 5.6% for those fractures with the teeth in the line of fracture extracted. PMID- 19304039 TI - An in vitro evaluation of rigid internal fixation techniques for sagittal split ramus osteotomies: advancement surgery. AB - PURPOSE: This in vitro investigation was developed with the purpose of comparing the biomechanical features of 3 different methods of rigid internal fixation for sagittal split ramus osteotomies for mandibular advancement. MATERIALS AND METHODS: Rigid internal fixation techniques included a 4-hole plate and 4 monocortical screws (miniplate group), a 4-hole plate and 4 monocortical screws with 1 additional bicortical positional screw (hybrid group), and 3 bicortical positional screws in a traditional inverted-L pattern (inverted-L group). Screws and miniplates were made of titanium and from a 2.0-mm system (MDT, Rio Claro, SP, Brazil). Sixty polyurethane replicas of human hemimandibles (Nacional, Jau, SP, Brazil) were used as substrates, simulating a 5-mm advancement surgery by a sagittal split ramus osteotomy. They were adapted to a test support, and were submitted to lateral torsional forces on the buccal molar surface and vertical cantilever loading on the incisal edge by an Instron 4411 mechanical testing unit (Instron, Norwood, MA) for recording peak loading at 1 mm, 3 mm, 5 mm, and 10 mm of displacement. Each group was formed by 10 replicas, subjected to a linear noncyclical testing only once. Means and standard deviation were analyzed using analysis of variance and Tukey tests, with a 5% level of significance. Testing failures were also recorded. RESULTS: The miniplate group showed lowest load peak scores (P < .01) when compared with the other fixation techniques, irrespective of the direction of force. The inverted-L group showed higher resistance (P < .01) than the hybrid group when vertical forces were applied. For molar load, the hybrid and inverted-L groups showed no significant difference. CONCLUSIONS: For mandibular advancement surgery of 5 mm, it was concluded that the rigid internal fixation technique for sagittal split ramus osteotomies based on 3 bicortical screws in the inverted-L pattern was the most stable in a laboratory environment. Furthermore, the results suggested that installation of a bicortical positional screw in the retromolar region may significantly optimize the resistance of the miniplate and monocortical screw fixation. PMID- 19304040 TI - Continuous versus discontinuous distraction: evaluation of bone regenerate following various rhythms of distraction. AB - PURPOSE: To investigate continuous distraction osteogenesis (DO) of the nasal bones in a rabbit model, and to compare data from this continuous DO study with data from a previously conducted discontinuous DO study. In addition, radiographic and ultrasonographic bone-fill scores were determined to investigate whether these scores provided reliable predictive value for the amount of new bone formation in the distraction area. MATERIALS AND METHODS: Skeletally mature female New Zealand White rabbits were subjected to distraction of the nasal bones. A custom-made continuous distractor was used to perform automatic non-stop distraction. Bone data were obtained from radiography, ultrasonography, and microcomputed tomography. Data from this experiment were compared with data from a previous study on discontinuous distraction rhythms. RESULTS: Ultrasonographic bone-fill scores correlated significantly to actual bone volume in contrast to radiographic bone-fill scores. Bone volume was significantly higher in the continuous DO group compared with the discontinuous DO groups. CONCLUSION: Continuous distraction resulted in accelerated osteogenesis compared with discontinuous distraction. Furthermore, bone-fill scores based on ultrasonography showed a significant correlation with actual bone volumes. PMID- 19304041 TI - Infection rates following perioperative prophylactic antibiotics versus postoperative extended regimen prophylactic antibiotics in surgical management of mandibular fractures. AB - PURPOSE: To determine whether or not an extended regimen of prophylactic antibiotics following either open or closed reduction of mandibular fractures is beneficial in lowering the rate of infection in postoperative patients. PATIENTS AND METHODS: This study is a retrospective chart review of 150 patients treated operatively for both complicated and uncomplicated mandibular fractures at University of New Mexico Health Sciences Center in Albuquerque, NM, between January 1, 2000 and June 12, 2007. Treatment modalities used were closed reduction with maxillomandibular fixation or open reduction and internal fixation with either wire osteosynthesis or rigid internal fixation. Patients fell into 1 of 2 groups: the first group received antibiotics perioperatively, which consisted of no more than 24 hours of antibiotics in the postoperative period; the second group received extended regimen antibiotics, which consisted of anywhere from 24 hours to 10 days of antibiotics in the postoperative period. Seventy-five patients were included in each group. The type of antibiotic prescribed was at the discretion of the operating surgeon. This study was conducted using an intention-to-treat analysis. Postoperative infection was diagnosed either subjectively or objectively by the clinician evaluating the patient at follow-up appointments. RESULTS: In the extended antibiotic group, 8 out of 75 subjects (10.67%) developed infection. In the perioperative antibiotic group, 10 out of 75 subjects (13.33%) developed infection. Statistical analysis using chi(2) distribution showed that this difference in proportions was not significant (chi(2) = 0.06, P = 0.8). CONCLUSION: This study found that the use of postoperative prophylactic antibiotics does not have a statistically significant effect on postoperative infection rates in surgical management of complicated or uncomplicated mandibular fractures. PMID- 19304042 TI - Assessment of pterygomaxillary separation in Le Fort I Osteotomy in class III patients. AB - PURPOSE: To examine the separation of the pterygomaxillary region at the posterior nasal spine level after Le Fort I osteotomy in Class III patients. PATIENTS AND METHODS: The study group consisted of 37 Japanese patients with mandibular prognathism and asymmetry, with maxillary retrognathism or asymmetry. A total of 74 sides were examined. Le Fort I osteotomy was performed without a pterygoid osteotome, with an ultrasonic curette used to remove interference at the pterygomaxillary region. Postoperative computed tomography (CT) was analyzed for all patients. The separation of the pterygomaxillary region and the location of the descending palatine artery were assessed. RESULTS: Although acceptable separation between the maxilla and pterygoid plates was achieved in all patients, an exact separation of the pterygomaxillary junction at the posterior nasal spine level was found in only 18 of 74 sides (24%). In 29 of 74 sides (39.2%), the separation occurred anterior to the descending palatine artery. In 29 of 74 sides (39.2%), complete separation between the maxilla and lateral and/or medial pterygoid plate was not achieved, but lower level separation of the maxilla and pterygoid plate was always complete. The maxillary segments could be moved to the postoperative ideal position in all cases. CONCLUSION: Le Fort I osteotomy without an osteotome does not always induce an exact separation at the pterygomaxillary junction at the posterior nasal spine level, but the ultrasonic bone curette can remove the interference between maxillary segment and pterygoid plates more safely. PMID- 19304043 TI - Surgical treatment of lip cancer: our experience with 106 cases. AB - PURPOSE: To report our experience with 106 cases of lip cancer. PATIENTS AND METHODS: We treated 106 patients with stages T1, T2, or T3 lip cancer (76, 22, and 8 cases, respectively). For the 34 T1 lesions up to 1 cm in diameter, we used a V or W excision. In the 42 T1 lesions greater than 1 cm and the 20 T2 lesions, we used the staircase technique. In 2 T2 cases, the carcinoma was located on the labial commissure and was treated with the Fries technique. For the 8 T3 cases, we used the Bernard-Freeman-Fries technique. In 28 patients, a lip shave was performed and tumor was removed. The 7 patients who were N+ at diagnosis underwent modified radical neck dissection and radiotherapy. RESULTS: Ten patients died during the follow-up period of 11 to 65 months: 8 of unrelated causes and 2 of new upper aerodigestive tract carcinoma. None of the patients died of their lip cancer. CONCLUSIONS: Lip cancer is a frequent disease of the oral cavity. Although general agreement has been reached concerning stage T and N+ surgical treatment, unresolved questions remain with regard to N0 treatment. We present our experience and suggestions. PMID- 19304044 TI - Radiographic morphometric study of 37 periapical lesions in 30 patients: validation of success criteria. AB - PURPOSE: The purpose of this study was to establish the midterm prognosis of periapical surgery by use of the ultrasound technique, comparing a scale obtained by image analysis with the success rates according to other prognostic scales. MATERIALS AND METHODS: A clinical follow-up study was performed between September 2004 and September 2005. Thirty-six patients with chronic periapical lesions were treated with periapical surgery via the ultrasound technique and root-end filling with silver amalgam. Evolution was analyzed by use of the different currently accepted assessment scales at 6 and 12 months after intervention with a scale obtained by radiographic image analysis. RESULTS: Thirty patients with a mean age of 40.5 years were studied. The mean follow-up period was 14.2 months. Thirty seven lesions were analyzed. A positive correlation at 6 months was observed between the radiographic scale proposed in this study and the scales of Rud and Andreasen and von Arx et al. At 12 months, the proposed radiographic scale only correlated with the functional tooth concept established by Friedman. CONCLUSIONS: There was no statistically significant relation between the radiographic scale from the image analyzer and the criteria of Rud and Andreasen and von Arx et al. PMID- 19304045 TI - Bisphosphonate-related osteonecrosis of the jaws: a single-center study of 101 patients. AB - PURPOSE: Osteonecrosis of the jaw (ONJ) is a devastating side effect of long-term bisphosphonate (BP) use. We present the largest case series from a single department. MATERIALS AND METHODS: This case series included 101 ONJ patients. Data on demographics, medical background, type and duration of BP use, possible triggering events, mode of therapy, and outcome were recorded. RESULTS: ONJ was associated with intravenous BPs in 85 patients and with oral BPs in 16 patients. It was diagnosed after 48, 27, and 67 months of pamidronate, zoledronic acid, and alendronate use, respectively. Long-term antibiotics and minimal surgical procedures resulted in complete or partial healing in 18% and 52% of the patients, respectively; 30% had no response. There was no association between ONJ and diabetes, steroid and antiangiogenic treatment, or underlying periodontal disease. Diagnostic biopsies aggravated lesions without being informative about pathogenesis. A conservative regimen is our treatment of choice. CONCLUSION: Solutions for decreasing morbidity and poor outcome of ONJ remain elusive. PMID- 19304046 TI - Mastication and late mandibular fracture after surgery of impacted third molars associated with no gross pathology. AB - PURPOSE: This study was undertaken with the null hypothesis that in patients, fully denate or with 1 or 2 teeth missing and older than 25 years, mastication does not affect late mandibular fracture after surgical removal of impacted third molars (M3s) associated with no gross pathology. MATERIALS AND METHODS: Five hundred sixty patients, fully dentate or with 1 or 2 teeth missing and older than 25 years who had no gross pathology associated with their impacted lower M3s, were recruited in this study. They were operated on under local anesthesia using a standard technique and randomly assigned into 2 groups for nonroutine (NR group) and routine (R group) postoperative instructions. In the NR group, patients were postoperatively educated in the possibility of mandibular fracture and were given an emphasis on the necessity of limiting mastication to a soft diet for 4 weeks. In the R group, patients were given no such education or emphasis. Patients were followed up for 2 months, and data concerning patients' age and gender; tooth position, angulation, and depth; date and site of surgery; and occurrence of late mandibular fracture were recorded and statistically analyzed. A value of P less than .05 was considered statistically significant. RESULTS: In no patient group was there a late mandibular fracture recorded. All patients completed the follow-up period, and most of the R group patients had normal eating habits 10 to 14 days after surgery. In no patient group was there a statistically significant difference in relation to gender (P = .735), site of surgery (P = .552), class horizontal space available (P = .427), class highest portion of the M3 crown (P = .424), angulations of the teeth (P = .925), and severity of impaction (P = .445). CONCLUSIONS: In patients, fully dentate or with 1 or 2 teeth missing and older than 25 years who have no jawbone atrophy and no systemic problems that may impair bone strength, mastication seems not to affect late mandibular fracture after surgical removal of impacted M3s associated with no gross pathology. The remote possible risk of the late fracture shown in our patients indicates the need for no special precautions. PMID- 19304047 TI - Radiolucent lesion of the posterior mandible. PMID- 19304048 TI - Methods to treat the edentulous posterior maxilla: implants with sinus grafting. AB - Prosthetic rehabilitation of the edentulous posterior maxilla with implant supported prostheses frequently presents a challenge for the oral surgeon because of the lack of bone due to alveolar ridge resorption or maxillary sinus pneumatization. To overcome these problems, different solutions were proposed over the years, such as the use of short implants or tilted implants (including zygoma implants), with the aim of avoiding maxillary sinus floor elevation. Both of these techniques have advantages and disadvantages that should be evaluated carefully to choose the most appropriate treatment. Zygoma implants or short/tilted implants are not a panacea for the treatment of patients with inadequate posterior maxillary bone stock. Instead, treatment should be based on the characterization of resorption patterns of the posterior maxilla, and may include the need for sinus grafting or other grafting procedures to reestablish not only adequate bone volume for implant placement, but also a favorable intermaxillary relationship, to optimize the functional and esthetic outcome of the final prosthetic rehabilitation. The authors discuss the indications, advantages, and disadvantages of sinus-grafting procedures in association with or without other reconstructive procedures. PMID- 19304049 TI - Nongrafting implant options for restoration of the edentulous maxilla. AB - Patients who need implant-supported prostheses in the maxilla might not have sufficient bone height in the posterior region for implants. Treatment alternatives in such cases include either bone graft augmentation followed by implants or nongrafting options. This report discusses 2 nongrafting options for prosthetic rehabilitation in these patients. An extensive summary of the published data has been performed to provide evidence for these recommendations. PMID- 19304050 TI - Management of atrophic edentulous mandibular fractures: the case for primary reconstruction with immediate bone grafting. PMID- 19304051 TI - Osteoid osteoma of subcondylar region: case report and review of the literature. PMID- 19304052 TI - Synovial chondromatosis of the temporomandibular joint: a case report with bilateral occurrence. PMID- 19304053 TI - Massive hemorrhage following bilateral sagittal split ramus osteotomy: a case report. PMID- 19304054 TI - Foreign body in the hard palate of children and risk of misdiagnosis: report of 3 cases. PMID- 19304055 TI - Orthodontic consequences of ritual dental mutilations in northern Tchad. PMID- 19304056 TI - Suppurative thrombophlebitis of the internal jugular vein: a rare complication of the mandible fracture. PMID- 19304057 TI - Clear cell sarcoma of the temporal region: case report, review of the literature, and genetic analysis. PMID- 19304058 TI - Synovial sarcoma of the tongue: case report and review of the literature. PMID- 19304059 TI - Myositis ossificans: a case report of multiple recurrences following third molar extractions and review of the literature. PMID- 19304060 TI - Median palatal cyst: case report and review of literature. PMID- 19304061 TI - Use of a sputum trap to facilitate pus specimen collection from an abscess cavity. PMID- 19304062 TI - Re: Tibial articular cartilage wear in varus osteoarthritic knees: correlation with cruciate ligament integrity and severe deformity (Mullaji et al.). PMID- 19304063 TI - Re: "Patellar tendon length following knee arthroplasty with and without preservation of the infrapatellar fat pad "(Lemon et al.). PMID- 19304064 TI - Post-cataract endophthalmitis: can we do better? PMID- 19304065 TI - Safety limits of online optical coherence pachymetry. PMID- 19304068 TI - Corneal power measurements after keratorefractive surgery. PMID- 19304069 TI - Retention of an anterior chamber IOL versus IOL exchange in endothelial keratoplasty. PMID- 19304070 TI - Cost-benefit analysis of multifocal IOLs versus monovision pseudophakia. PMID- 19304072 TI - Consultation Section. Cataract surgical problem. PMID- 19304080 TI - Collagen crosslinking with ultraviolet-A and hypoosmolar riboflavin solution in thin corneas. AB - Corneal collagen crosslinking (CXL) with riboflavin and ultraviolet-A light is a method for treating progressive keratectasia. The currently accepted treatment parameters induce collagen crosslinking in the anterior 250 to 350 microm of corneal stroma. To protect the endothelium, CXL inclusion criteria require a minimum corneal thickness of 400 microm after removal of the epithelium. In advanced keratoconus, however, progressive corneal thinning often leads to a remaining stromal thickness of less than 400 microm. We have therefore modified the current treatment protocol by preoperatively swelling thin corneas to a stromal thickness of at least 400 microm using hypoosmolar riboflavin solution. This treatment protocol was performed in a case series of 20 patients, and no complications were observed. Preoperative swelling of the cornea safely broadens the spectrum of CXL indications to thin corneas that would otherwise not be eligible for treatment. PMID- 19304081 TI - Simple technique for graft insertion in Descemet-stripping (automated) endothelial keratoplasty using a 30-gauge needle. AB - We describe a needle insertion technique for graft insertion in Descemet stripping (automated) endothelial keratoplasty (DSEK/DSAEK). A folded donor posterior corneal disk is inserted through a 5.0 mm scleral tunnel incision over a plastic glide using a 30-gauge needle. The technique enables safe and easy graft insertion without vertical or horizontal compression of the donor tissue, causing minimal trauma to the donor endothelium and/or host structures. PMID- 19304082 TI - Incidence of post-cataract endophthalmitis at Aravind Eye Hospital: outcomes of more than 42,000 consecutive cases using standardized sterilization and prophylaxis protocols. AB - PURPOSE: To report the incidence of postoperative endophthalmitis at a high volume eye hospital in southern India using a modified cost-effective sterilization protocol. SETTING: Aravind Eye Hospital and Post Graduate Institute of Ophthalmology, Pondicherry, India. METHODS: In this retrospective observational series at a single eye hospital, records of patients who had cataract surgery using a modified sterilization protocol from January 2007 through August 2008 and developed postoperative endophthalmitis within the first 3 postoperative months were drawn from a computerized database. The patient's socioeconomic status, the surgeon's experience, and the type of cataract procedure performed were analyzed as possible risk factors using the chi-square test/Fischer exact test. RESULTS: During the study period, 42426 cataract surgeries were performed. From these, 38 cases of presumed postoperative endophthalmitis were identified (incidence 0.09%). Thirty-five of the 38 cases were in the manual large- and small-incision extracapsular cataract extraction (ECCE) group, which had a statistically higher rate than the phacoemulsification group (P = .016). There was no statistical difference in the endophthalmitis rates between private patients and charity patients for either surgical method (manual ECCE or phacoemulsification). CONCLUSIONS: The modified sterilization and asepsis protocol adopted to facilitate high-volume cataract surgery in a clinical setting appeared to be safe and effective in preventing postsurgical endophthalmitis. Despite a 3:1 ratio of manual ECCE to phacoemulsification and the elimination of certain traditional sterilization practices, the rate of endophthalmitis in this generally underserved patient population with multiple risk factors for infection was comparable to that reported in other modern settings. PMID- 19304083 TI - Prophylactic intracameral cefazolin after cataract surgery: endophthalmitis risk reduction and safety results in a 6-year study. AB - PURPOSE: To assess the use of intracameral cefazolin in preventing endophthalmitis in cataract surgery. SETTING: Ophthalmology Department, L'Hospitalet de Llobregat, Barcelona, Spain. METHODS: This study was of phacoemulsification procedures performed from January 2002 to December 2007. In January 2004, intracameral cefazolin given at the end of the surgery was added to the prophylaxis protocol of cataract surgery. The cumulative incidence of postoperative endophthalmitis before and after the addition of intracameral cefazolin was compared. RESULTS: During the study period, 18579 phacoemulsification procedures were performed. In the 2-year period before introduction of intracameral cefazolin prophylaxis, 25 cases of endophthalmitis were diagnosed in 5930 surgeries, leading to a cumulative incidence of 0.422% (95% confidence interval [CI], 0.279%-0.613%). After the introduction of cefazolin, 6 cases of endophthalmitis were diagnosed in 12649 surgeries, an incidence of 0.047% (95% CI, 0.019%-0.099%). When only microbiologically proven cases were considered, the cumulative endophthalmitis incidence was 0.388% (95% CI, 0.252%-0.572%) in the first study period and 0.032% (95% CI, 0.010%-0.076%) in the second study period (P<.0000001). The relative risk for presenting with endophthalmitis in the first study period compared with the second period was 8.89 (95% CI, 3.65-21.65). CONCLUSIONS: A 2.5 mg/0.1 mL intracameral bolus of cefazolin provided excellent prophylactic effectiveness, with a reduction in the incidence of endophthalmitis from 0.422% to 0.047%, corresponding to a relative risk reduction of 88.7% (95% CI, 72.6%-95.4%). Cefazolin fulfills international recommendations on antimicrobial prophylaxis for surgical site infections and is easier to obtain in developing countries. PMID- 19304084 TI - Acute-onset nosocomial endophthalmitis after cataract surgery: incidence, clinical features, causative organisms, and visual outcomes. AB - PURPOSE: To estimate the incidence of acute-onset endophthalmitis after cataract surgery and to report its clinical features, microbiology, and final visual outcomes. SETTING: King Khaled Eye Specialist Hospital, Riyadh, Saudi Arabia. METHODS: This was a retrospective observational case series. RESULTS: The 10-year incidence of acute-onset endophthalmitis after cataract surgery was 0.068%: 0.049% for extracapsular cataract extraction (ECCE) and 0.085% for clear corneal phacoemulsification (P = .268). The incidence of endophthalmitis was 0.053% during the first 5 years of the study and 0.08% during the next 5 years. The most common presenting features were pain and poor red reflex. Of the 20 cases, 18 were culture positive. The culture isolates showed Staphylococcus species in 7 eyes (35%), Streptococcus species in 7 eyes (35%), polymicrobial or mixed infections in 3 eyes (15%), and Propionibacterium acnes in 1 eye (5%). Two patients (10%) achieved a final visual acuity of 20/40 or better, 5 (25%) had worse than 20/40 to better than 20/200, and 8 (40%) had worse than 20/200; 5 eyes (25%) were eviscerated. Visual outcomes were good in endophthalmitis cases after phacoemulsification and in cases caused by Staphylococcus epidermidis and worse in cases that had Streptococcus species endophthalmitis. CONCLUSIONS: Clear corneal phacoemulsification had a 1.73-fold higher risk for acute endophthalmitis than ECCE. Although the incidence of endophthalmitis after ECCE was lower than after phacoemulsification, the outcome was much worse in post-ECCE endophthalmitis cases. Poor visual outcomes were associated with more virulent organisms and delayed presentation. PMID- 19304085 TI - Comparison of ray-tracing method and thin-lens formula in intraocular lens power calculations. AB - PURPOSE: To compare the accuracy of the thin-lens and ray-tracing methods in intraocular lens (IOL) power calculations in normal eyes and eyes after corneal refractive surgery. SETTING: International Vision Correction Research Centre, University of Heidelberg, Heidelberg, Germany. METHODS: Pseudophakic eye models were constructed using Zemax optical software, importing corneal radii (normal ray tracing) and corneal surface elevation data (individual ray tracing) measured by Pentacam Scheimpflug photography. Algorithms to predict IOL position (effective lens position [ELP]) or postoperative anterior chamber depth [ACD(post)]) (Haigis, Hoffer Q, Norrby, Olsen 2) were used in the thin-lens and ray-tracing methods. Intraocular lens power was calculated in 25 eyes after corneal refractive surgery using normal and double-K modified thin-lens and ray tracing methods. RESULTS: Back-calculation of ELP and ACD(post) were well correlated. Using algorithms of Haigis, Hoffer Q, Norrby, and Olsen 2 to predict IOL position, mean absolute prediction errors (MAEs) of the thin-lens formula were 0.64 diopters (D) +/- 0.52 (SD), 0.57 +/- 0.46 D, 0.59 +/- 0.42 D, and 0.61 +/- 0.47 D, respectively; MAEs of normal ray-tracing method were 0.64 +/- 0.50 D, 0.58 +/- 0.44 D, 0.59 +/- 0.41 D, and 0.62 +/- 0.45 D, respectively; MAEs of individual ray-tracing method were 0.66 +/- 0.52 D, 0.59 +/- 0.45 D, 0.59 +/- 0.43 D, and 0.62 +/- 0.50 D, respectively. No statistical differences were found between the thin-lens and ray-tracing methods. CONCLUSION: Theoretical thin-lens formulas were as accurate as the ray-tracing method in IOL power calculations in normal eyes and eyes after refractive surgery. PMID- 19304086 TI - Wavefront aberrations, depth of focus, and contrast sensitivity with aspheric and spherical intraocular lenses: fellow-eye study. AB - PURPOSE: To compare wavefront aberration, depth of focus, contrast sensitivity, and in vivo modulation transfer function (MTF) after fellow-eye implantation of aspheric and spherical intraocular lenses (IOLs). SETTINGS: Department of Ophthalmology, St. Thomas' Hospital, London, United Kingdom. METHODS: This prospective randomized controlled study comprised patients with bilateral cataract who received an aspheric AcrySof SN60WF IOL or a spherical AcrySof SN60AT IOL in the first eye and the other IOL in the second eye. Assessments at 3 and 6 months included 100% and 9% logMAR best corrected visual acuity (BCVA) and photopic and mesopic functional acuity contrast testing. Total internal and corneal aberrations and depth of focus were computed. Distance-corrected near logMAR acuity was available at 12 months. RESULTS: At 3 and 6 months, there was no significant difference in 100% and 9% BCVA or photopic contrast sensitivity. Mesopic contrast sensitivity was better and total and internal spherical aberrations were significantly less with the aspheric IOL. Total and internal eye vertical coma was reduced with aspheric IOL. Total MTF was not significantly different between groups. The aspheric IOL group had 0.46 diopter less depth of focus than the spherical IOL group at 6 months (P<.05). Distance-corrected near acuity was significantly better with the spherical IOL. CONCLUSIONS: Aspheric IOLs significantly reduced spherical aberration, improving mesopic contrast sensitivity. Vertical coma was reduced with aspheric IOLs. Reduction of aberrations may be responsible for reduced depth of focus with aspheric IOLs. This may be disadvantageous for near vision and reading ability. PMID- 19304087 TI - Visual acuity comparison of 2 models of bifocal aspheric intraocular lenses. AB - PURPOSE: To compare visual acuity after bilateral implantation of 2 models of multifocal aspheric intraocular lenses (IOLs). SETTING: Fernandez-Vega Ophthalmological Institute, Oviedo, and University of Valencia, Valencia, Spain. METHODS: Prospective study of patients who had bilateral implantation of an AcrySof ReSTOR SN6AD3 IOL (Group 1) or an Acri.LISA 366D IOL (Group 2). Six months postoperatively, binocular uncorrected and best corrected distance visual acuity, uncorrected-distance and best distance-corrected near visual acuity, best corrected intermediate visual acuity, and the defocus curve were measured in both IOL groups. RESULTS: Group 1 comprised 36 eyes (18 patients) and Group 2, 40 eyes (20 patients). The mean binocular values in Group 1 and Group 2, respectively, were as follows: best corrected distance acuity, -0.05 +/- 0.09 logMAR and -0.08 +/- 0.08 logMAR (both approximately 20/20); best distance-corrected near acuity, 0.01 +/- 0.16 logMAR and -0.05 +/- 0.07 logMAR; best corrected intermediate acuity at 80 cm, 0.20 +/- 0.18 logMAR (approximately 20/32) and 0.16 +/- 0.13 logMAR (approximately 20/25) and at 60 cm, 0.16 +/- 0.16 logMAR (approximately 20/25) and 0.18 +/- 0.13 logMAR (approximately 20/25). There were no statistically significant between-group differences in visual acuity at any distance (P>.3). Defocus curves were similar between groups (2.00 to -5.00 diopters) (P>.26). CONCLUSIONS: The 2 multifocal aspheric IOL models gave similar and good high-contrast visual acuity at distance and near. Intermediate visual acuity, also comparable between IOL models, was better than published results of a spherical IOL model. PMID- 19304088 TI - Objective assessment of inflammation after cataract surgery: comparison of 3 similar intraocular lens models. AB - PURPOSE: To compare postoperative inflammation in patients receiving 1 of 3 AcrySof intraocular lenses (IOLs): MA60AC (Group 1), SA60AT (Group 2), or SN60AT (Group 3). SETTING: Service d'Ophtalmologie, Universite Paris Descartes Hopital Cochin, Paris, France. METHODS: This prospective randomized 3-month study included eyes that received 1 of the 3 IOL models with standard surgery and postoperative care. Anterior chamber cells were assessed at the slitlamp and anterior chamber flare values, with a Kowa 500 flare meter. RESULTS: Fifty-nine patients (59 eyes) with a mean age of 72.7 years were evaluated. Group 1 and Group 2 comprised 20 eyes each and Group 3, 19 eyes. No eye had anterior chamber cells at baseline. In all 3 groups, the presence of anterior chamber cells was highest 1 week postoperatively and generally decreased at subsequent visits, with no statistically significant differences between IOL groups at 1 week (P = .2655), 1 month (P = .073), or 3 months (P = .5766). A similar proportion of eyes in each IOL group had residual cells in the anterior chamber at 3 months; the cells were not clinically significant. In all groups, the mean flare values were low (<11 photons/ms) at baseline (P = .4522) and statistically similar between groups at each subsequent visit (P>or=.2801). There were no adverse events. CONCLUSION: The 3 IOLs models, including the blue light-filtering model, had similar anterior chamber cells and flare values over a 3-month period, showing the lack of difference in inflammation induced by cataract surgery with implantation of the 3 similar IOL models. PMID- 19304089 TI - Mitomycin-C in hyperopic photorefractive keratectomy. AB - PURPOSE: To evaluate the effects of topical mitomycin-C (MMC) after hyperopic photorefractive keratectomy (PRK). SETTING: Private practice, Siena, Italy. METHODS: In this prospective study, a sponge with 0.2 mg/mL MMC or balanced salt solution (controls) was placed on the stroma for 45 seconds after PRK performed using a Technolas 217z laser. After epithelialization, fluorometholone 0.1% eyedrops were used for 4 months in both groups. RESULTS: The MMC group comprised 88 eyes (mean spherical equivalent [SE] +3.51 diopters [D] +/- 1.04) and the control group, 91 eyes (mean SE +3.50 +/- 1.03 D). At 18 months, the mean SE was 0.10 +/- 0.37 D and 0.22 +/- 0.70 D, respectively, and the mean defocus equivalent, 0.34 +/- 0.32 D and 0.69 +/- 0.74 D, respectively (both P<.05). The mean best spectacle-corrected visual acuity (BSCVA) was 0.06 +/- 0.08 logMAR in the MMC group and 0.08 +/- 0.10 logMAR in the control group; 2 eyes and 11 eyes, respectively, lost more than 0.1 logMAR of BSCVA. The mean uncorrected visual acuity (UCVA) was 0.13 +/- 0.11 logMAR in the MMC group and 0.21 +/- 0.20 logMAR in the control group (P<.05). The UCVA was better than 20/40 in 94% of eyes and 80% of eyes, respectively. The efficacy index was 0.87 and 0.67, respectively. Haze at 18 months was 0.05 +/- 0.11 in the MMC group and 0.23 +/- 0.46 in the control group (P<.05). No endothelial damage was observed in either group. CONCLUSION: Mitomycin-C prevented haze formation and improved predictability and efficacy. No adverse effects occurred. PMID- 19304090 TI - Effect of anterior and posterior corneal surface irregularity on vision after Descemet-stripping endothelial keratoplasty. AB - PURPOSE: To evaluate irregularity of the anterior and posterior cornea before and after Descemet-stripping endothelial keratoplasty (DSEK) and its effect on visual acuity. SETTING: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan. METHODS: Corneal data were acquired using a rotating Scheimpflug camera before and 1 month and 3 months after DSEK. Anterior and posterior corneal elevation data were decomposed into a set of Zernike polynomials up to the 8th order within a 4.0 mm diameter region. Total higher-order root mean square (HO RMS) and RMS from the 3rd to 8th order were calculated. The effects of anterior and posterior surface irregularity on visual acuity were evaluated. RESULTS: This clinical study comprised 13 consecutive eyes of 12 bullous keratopathy patients. The mean best corrected visual acuity (BCVA) was 1.11 logMAR +/- 0.5 (SD) preoperatively, 0.49 +/- 0.49 logMAR 1 month postoperatively, and 0.27 +/- 0.32 logMAR at 3 months. The HO-RMS of the anterior surface 1 month (P = .040) and 3 months (P = .048) postoperatively was significantly lower than preoperatively. There were no significant differences in posterior surface HO-RMS between preoperatively and 1 month (P = .45) and 3 months (P = .054). The postoperative BCVA was significantly correlated with HO-RMS (P<.001), but not with posterior surface HO-RMS, at 3 months (P = .354). CONCLUSION: Postoperative BCVA correlated with irregularity of the anterior surface but not the posterior surface. In addition to corneal transparency, regularity of the anterior surface is an important factor in visual acuity after DSEK. PMID- 19304091 TI - Wavefront aberrations in eyes with decentered ablations. AB - PURPOSE: To compare the refractive and functional outcomes and wavefront profiles in eyes with decentered ablations and eyes with well-centered ablations. SETTING: Medical Research Foundation, Chennai, Tamil Nadu, India. METHODS: This retrospective analysis comprised eyes with topographically diagnosed decentered ablations after laser in situ keratomileusis (LASIK). Refraction, contrast sensitivity, and ocular wavefront aberrations were measured preoperatively and 1 month postoperatively. The induced aberrations in these eyes were compared with those in eyes with well-centered ablations. RESULTS: Forty-six eyes (38 patients) had decentered ablations and 60 eyes (32 patients), well-centered ablations. The mean decentration in the study group was 0.86 mm +/- 0.29 (SD) (range 0.35 to 1.61 mm). There was no significant correlation between decentration and attempted refractive correction. There was, however, a statistically significant (P<.05) linear correlation between the distance of decentration and the magnitude of induced tilt (r = -0.31), coma (r = -0.41), and secondary astigmatism (r = 0.36). The induced changes in tilt, oblique astigmatism, vertical coma, and spherical aberration were statistically significantly higher in eyes with decentered ablations than in eyes with well-centered ablations. A statistically significantly higher percentage of eyes (87%) with well-centered ablations than eyes with decentered ablations (70%) had a postoperative uncorrected visual acuity (UCVA) of 20/20 or better. There was no significant difference in contrast sensitivity between groups. CONCLUSION: Eyes with decentered ablations had a significantly higher magnitude of induced aberrations and lower UCVA than eyes with well-centered ablations. PMID- 19304092 TI - Comparison of early postoperative clinical outcomes of photorefractive keratectomy and lamellar epithelial debridement. AB - PURPOSE: To compare early postoperative clinical outcomes of photorefractive keratectomy (PRK) and lamellar epithelial debridement (LED). SETTING: Department of Ophthalmology, Kangbuk Samsung Hospital, Seoul, Korea. METHODS: This prospective study was of patients randomly assigned to have PRK or LED. In the LED group, an epithelial flap was created using an Amadeus II epikeratome. Postoperative follow-up was at 1, 3, and 7 days and 1, 3, and 6 months. The outcome parameters were uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), higher-order aberrations (HOAs), epithelial healing time, and corneal haze. RESULTS: The study comprised 39 patients (76 eyes). The mean preoperative spherical equivalent (SE) was -3.96 diopters (D) +/- 1.24 (SD) in the PRK group and -4.06 +/- 1.39 D in the LED group. Postoperative UCVA was significantly better in the LED group 1 day postoperatively. The UCVA was 20/20 or better in 14.6% in the PRK group and 42.9% in the LED group (P = .006); 20/25 or better in 41.5% and 82.9%, respectively (P = .000); and 20/40 or better in 80.5% and 100%, respectively (P = .006). On subsequent follow-up visits, the UCVA was comparable between groups. No eye lost lines of BCVA at 3 months. There was no difference between groups in postoperative SE refraction, HOAs, or corneal haze. CONCLUSIONS: Lamellar epithelial debridement and PRK had comparable safety and efficacy in the surgical correction of low to moderate myopia. The UCVA was significantly better after LED than after PRK 1 day postoperatively and equivalent thereafter. PMID- 19304093 TI - Assessment of visual performance in pseudophakic monovision. AB - PURPOSE: To assess the visual performance and acceptability of pseudophakic monovision and examine its relationship to age. SETTING: Department of Ophthalmology, Kitasato University Hospital, Kanagawa, Japan. METHODS: This retrospective study examined patients with pseudophakic monovision using monofocal intraocular lenses. Refractive errors, visual acuity at various distances, contrast sensitivity, and near stereopsis were measured. Patient satisfaction with monovision was evaluated using a questionnaire. RESULTS: Eighty two patients (age 49 to 87 years) were evaluated. The mean difference in spherical equivalent refractive error between each patient's eyes was 2.27 diopters. Most patients had a binocular uncorrected visual acuity of 0.10 logMAR or better at all distances. For contrast sensitivity, binocular summation was observed at 1.5 to 6.0 cycles per degree. Near stereopsis was in the normal range, which was up to 100 seconds of arc. Questionnaire responses showed that 81% of patients (64% <60 years; 87% between 60 years and 70 years; 94% older than 70 years) were satisfied with the results. CONCLUSION: Pseudophakic monovision was an effective approach for managing loss of accommodation after cataract surgery in patients older than 60 years; however, a careful selection process is required. PMID- 19304094 TI - Effect of diabetes mellitus on biomechanical parameters of the cornea. AB - PURPOSE: To compare parameters of biomechanical response of the human cornea measured as corneal hysteresis (CH) and corneal resistance factor (CRF) in patients with diabetes mellitus and healthy control subjects. SETTING: Department of Ophthalmology, Assaf Harofeh Medical Center, Zerifin, Israel. METHODS: In the right eye of each participant, the CH, CRF, Goldmann-correlated intraocular pressure (IOPg), and corneal-compensated intraocular pressure (IOPcc) were measured with the Ocular Response Analyzer. Central corneal thickness (CCT) was measured by ultrasonic pachymetry and intraocular pressure by Goldmann applanation tonometry (IOP GAT). Findings were compared between the 2 groups (control and diabetic). RESULTS: Forty diabetic patients (17 women, 23 men) and 40 healthy subjects (19 women, 21 men) were prospectively recruited. The mean CH was 9.3 mm Hg +/- 1.4 (SD) and 10.7 +/- 1.6 mm Hg and the mean CRF was 9.6 +/- 1.6 mm Hg and 10.9 +/- 1.7 mm Hg in the control group and diabetic group, respectively (both P < .0001). Diabetic corneas were significantly thicker (P = .019); the mean CCT was 530.3 +/- 35.9 microm in the control group and 548.7 +/- 33.0 microm in the diabetic group. The CH and CRF remained significantly different in multivariate analysis that included CCT. There was no statistically significant difference between the 2 groups in IOPcc, IOPg, or IOP GAT measurements. CONCLUSIONS: Diabetes mellitus affected biomechanical parameters of the human corneas, including increased CH, CRF, and CCT. Whether this observation has implications in the clinical management and understanding of corneal ectasia and glaucoma requires further study. PMID- 19304095 TI - Management of congenital cataract in children younger than 1 year using a 25 gauge vitrectomy system. AB - PURPOSE: To evaluate the safety and efficacy of a 25-gauge vitrectomy system for the management of congenital cataract in children younger than 1 year. SETTING: Princess Margaret Hospital for Children, Western Australia, Australia. METHODS: Children in a tertiary pediatric hospital and private practice had cataract extraction using a 25-gauge vitrectomy system between January 2005 and June 2008. Each eye had anterior vitrectorhexis, lens aspiration, posterior vitrectorhexis, and anterior vitrectomy through two 25-gauge limbal side ports. The ports were created with a 25-gauge trocar or a 0.6 mm paracentesis knife. The eyes were left aphakic. RESULTS: Nineteen (95%) of the 20 eyes (14 children) had a successful surgical outcome with the 25-gauge vitrectomy system. One eye was converted to the 20-gauge system because of an unusually tough and fibrous lens capsule. The limbal side ports in 16 of 17 eyes created with the 25-gauge trocar required suturing to seal the ports; none of the 3 eyes with side ports created with the paracentesis knife required suturing. There were no significant intraoperative or postoperative complications other than ocular hypertension in 1 eye. CONCLUSIONS: The 25-gauge vitrectomy system appears safe and effective for the management of infantile cataract. Advantages include more precise manipulations with smaller instruments in infant eyes, a more stable anterior chamber, and less postoperative astigmatism. PMID- 19304096 TI - Outcomes of cataract surgery in children with chronic uveitis. AB - PURPOSE: To evaluate the outcomes of cataract surgery in children with chronic uveitis. SETTING: Massachusetts Eye Research and Surgery Institution, Boston, Massachusetts, USA. METHODS: This retrospective chart review was of patients younger than 17 years with a history of uveitis who had cataract surgery before June 2004. RESULTS: Thirty-four children (41 eyes) were identified. The mean age of the 10 boys and 24 girls was 9.8 years (range 4 to 17 years) and the mean total follow-up, 4.1 years (range 0.3 to 15.7 years). Twenty-one children had juvenile idiopathic arthritis-associated uveitis, 7 had pars planitis, and 6 had other conditions. Sixteen patients had concomitant posterior segment pathology, 25 received perioperative immunomodulatory therapy, and 13 had intraocular lens (IOL) implantation. The postoperative best corrected visual acuity improved in 35 of 41 eyes; 31 eyes had an improvement of 3.6 lines at 1 year. Most patients (92%) improved after IOL implantation. Most patients (88%) who received immunomodulatory therapy attained better vision, but this was not statistically significant compared with those who did not (P = .47). Similarly, there was no statistically significant difference between those with posterior pathology and those without. At the end of the analysis (1 year), the cumulative probability of improvement in visual acuity in 41 eyes reached 0.91. CONCLUSION: In most cases, and with optimum control of intraocular inflammation, cataract surgery improved the visual outcome in children with chronic uveitis. Intraocular lens implantation was well tolerated in most cases, which may result in optimal vision. PMID- 19304097 TI - Effect of hydrodynamic parameters on corneal endothelial cell loss after phacoemulsification. AB - PURPOSE: To evaluate the effect of power, vacuum, and flow rate on endothelial cell loss after phacoemulsification. SETTING: Labbafinejad Medical Center Medical Center, Tehran, Iran. METHODS: In a prospective randomized clinical trial, phacoemulsification was performed in 2 groups (high vacuum and low vacuum) with 3+ nuclear sclerosis. The stop-and-chop technique was used with the Sovereign machine. Machine parameters during the chop stage were vacuum 400 mm Hg in the high-vacuum group and 200 mm Hg in the low-vacuum group and flow rate, 40 cc/min and 20 cc/min, respectively. Endothelial cell density preoperatively before and 1, 6, and 12 weeks postoperatively were compared. RESULTS: Each group comprised 30 eyes. The mean US power was 9.2% +/- 4.3% (SD) in the low-vacuum group and 13.1% +/- 4.6% in the high-vacuum group (P = .001) and the mean phaco time, 1.28 +/- 1.0 minutes and 0.88 +/- 0.6 minutes, respectively (P = .04). Total US energy and total fluid consumed were similar between groups. After 12 weeks, the mean endothelial cell loss was 9.0% +/- 4.0% in the low-vacuum group and 9.6% +/- 4.6% in the high-vacuum group (P = .6). There was a relationship between total US energy and endothelial loss (P<.001); however, total fluid volume was not a significant predictor (P = .19). CONCLUSIONS: Vacuum level did not have a significant effect on total US energy or total fluid consumed during phacoemulsification. There was a strong relationship between total US energy and endothelial cell loss but not between total infused fluid and endothelial cell loss. PMID- 19304098 TI - Effect of time sequences in scanning algorithms on the surface temperature during corneal laser surgery with high-repetition-rate excimer laser. AB - PURPOSE: To investigate the influence of temporal and spatial spot sequences on the ocular surface temperature increase during corneal laser surgery with a high repetition-rate excimer laser. SETTING: Institute for Refractive and Ophthalmic Surgery, Zurich, Switzerland, and WaveLight AG, Erlangen, Germany. METHODS: An argon-fluoride excimer laser system working at a repetition rate of 1050 Hz was used to photoablate bovine corneas with various myopic, hyperopic, and phototherapeutic ablation profiles. The temporal distribution of ablation profiles was modified by 4 spot sequences: line, circumferential, random, and an optimized scan algorithm. The increase in ocular surface temperature was measured using an infrared camera. RESULTS: The maximum and mean ocular surface temperature increases depended primarily on the spatial and temporal distribution of the spots during photoablation and the amount of refractive correction. The highest temperature increases were with the line and circumferential scan sequences. Significant lower temperature increases were found with the optimized and random scan algorithms. CONCLUSIONS: High-repetition-rate excimer laser systems require spot sequences with optimized temporal and spatial spot distribution to minimize the increase in ocular surface temperature. An ocular surface temperature increase will always occur depending on the amount of refractive correction, the type of ablation profile, the radiant exposure, and the repetition rate of the laser system. PMID- 19304099 TI - Modulation transfer function: rigid versus foldable phakic intraocular lenses. AB - PURPOSE: To study in a standard eye model the changes in modulation transfer function (MTF) of a monofocal intraocular lens (IOL) when a phakic IOL (pIOL) is placed in the anterior chamber, compare the MTFs of the rigid Artisan pIOL and foldable Artiflex pIOL, and evaluate the temporal evolution of the MTF of the foldable pIOL after the mechanical stress the pIOL undergoes when injected. SETTING: Fundacion Oftalmologica del Mediterraneo, Valencia, Spain. METHODS: The MTF values of the IOLs were calculated from the cross-line spread function recorded with the Opal Vector System. The measurements were taken using an eye model following the British and EN-ISO standards with 2.0 mm, 3.0 mm, 4.0 mm, and 5.0 mm pupils. A 28.00 diopter (D) Ophtec monofocal IOL was used as the crystalline lens. The 2 pIOLs were -9.00 D. RESULTS: The MTF of the rigid pIOL was slightly better than the MTF of the foldable pIOL with all pupil sizes. Both pIOLs provided good optics quality when compared with the monofocal IOL. The injection effect of the foldable IOL disappeared after 2 hours. CONCLUSIONS: The MTF of the monofocal IOL was slightly reduced with implantation of a negative pIOL in the anterior chamber. The rigid pIOL provided better optical performance than the foldable pIOL with all pupil sizes, as shown by the MTF values. The decrease in MTF caused by the mechanical stress on the foldable pIOL was nullified after 2 hours with no effect on optical quality. PMID- 19304100 TI - Induction of apoptosis of rabbit corneal endothelial cells by preservative-free lidocaine hydrochloride 2%, ropivacaine 1%, or levobupivacaine 0.75%. AB - PURPOSE: To determine and compare the amount of apoptosis and changes in rabbit corneal endothelial cell morphology after intracameral administration of different anesthetic agents. SETTING: Department of Ophthalmology, Baskent University Medical Faculty, Ankara, Turkey. METHODS: Right eyes of 64 Vienna white rabbits were injected intracamerally with preservative-free lidocaine hydrochloride 2%, ropivacaine 1%, levobupivacaine 0.75%, or fortified balanced salt solution (BSS Plus) (control). Animals were humanely killed 1 day or 7 days later. Terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling was used to detect apoptosis. Corneal endothelial cells and apoptotic cells were counted by light microscopy. The morphologic appearance was determined by transmission electron microscopy (TEM). RESULTS: Apoptotic cell density was high in the anesthetic groups on day 1 (P<.01); there was no significant difference between groups at 7 days. Apoptotic cell density declined significantly between 1 day and 7 days in the anesthetic groups (P<.05) but not in the control group. There was no difference in endothelial cell density between the 4 groups at 1 or 7 days. All anesthetic groups showed degenerative changes on TEM, with the least change in the preservative-free lidocaine hydrochloride 2% group. CONCLUSIONS: Intracameral injections of preservative-free lidocaine, ropivacaine, and levobupivacaine induced significantly more apoptotic endothelial cell loss than BSS Plus and led to morphologic changes in the corneal endothelial cells in the early period. This effect was temporary, with recovery by 7 days. Considering the limited proliferative capacity in human eyes, the induced apoptosis might result in the permanent cell loss and enlargement in human corneal endothelium. PMID- 19304101 TI - Intraocular lens power calculation after previous laser refractive surgery. AB - Methods to attempt more accurate prediction of intraocular lens power in refractive surgery eyes are many, and none has proved to be the most accurate. Until one is identified, a spreadsheet tool is available and can be used. It automatically calculates all the methods for which data are available on a single sheet for the patient's chart. The various methods and how they work are described. PMID- 19304102 TI - Endophthalmitis rates after implantation of the intraocular Collamer lens: survey of users between 1998 and 2006. AB - An anonymous on-line survey was sent to 234 intraocular Collamer lens (ICL) (Staar Surgical) surgeons in 21 countries to determine how many of their ICL cases had been complicated by endophthalmitis between January 1998 and December 2006. A second questionnaire about the infection details and treatment outcome was sent to those who reported cases of endophthalmitis. Ninety-five (40%) surgeons responded to the survey. They had implanted 17954 ICLs during the study period. Three surgeons reported 1 case of endophthalmitis each, a rate of 0.0167% (95%) confidence interval 0 to 0.036%) or approximately 1 case of endophthalmitis per 6000 ICL implantations. Follow-up details were available in 2 cases. Staphylococcus epidermidis was cultured in both cases, and both were treated without loss of vision. Endophthalmitis may be less common after ICL implantation than after cataract surgery. Further studies are required to define the rate and prognosis for endophthalmitis after ICL implantation to assist in accurate preoperative patient counseling. PMID- 19304103 TI - United Kingdom survey of antibiotic prophylaxis practice after publication of the ESCRS Endophthalmitis Study. AB - The European Society of Cataract & Refractive Surgeons Endophthalmitis Study published preliminary results in 2006 showing a near 5-fold decrease in the rates of postoperative endophthalmitis with the use of intracameral cefuroxime. The study findings have generated considerable controversy, and 1 year later its recommendations had been heeded by only 6% of American Society of Cataract and Refractive Surgery (ASCRS) members. This poll sought to gauge the uptake among surgeons in the United Kingdom and survey their response to its findings. Fifty five percent of respondents were using intracameral cefuroxime; 48% had switched after publication of the ESCRS study. Of those remaining, 68% reported their main concern was the risk for dilution errors in the absence of a commercially preformulated preparation, with 67% stating they would switch if such a product became available. Sixty-eight percent considered the lack of a subconjunctival cefuroxime treatment arm within the study a major methodology flaw. Sixty-seven percent said they do not consider penicillin allergy a contraindication to intracameral cefuroxime use, with the remaining 33% opting for alternative antibiotic prophylaxis. The majority of United Kingdom and Ireland Society of Cataract and Refractive Surgeons members have switched to intracameral cefuroxime, despite the absence of previously preferred practice in the study design. The major barrier to its further use appears to be the lack of a commercially available preformulated preparation. PMID- 19304104 TI - Case of late-onset corneal decompensation after iris-fixated phakic intraocular lens implantation. AB - A 48-year-old myopic patient with bilateral anterior chamber depth of 3.1 mm and endothelial cell density (ECD) of 2525 cells/mm(2) and 2638 cells/mm(2) preoperatively had bilateral implantation of an Artisan iris-fixated phakic intraocular lens (pIOL). Five years postoperatively, unilateral corneal stromal edema was seen in a circumscribed area overlying the temporal ridge of the pIOL in the right eye; the ECD was 1631 cells/mm(2) and the pachymetry, 586 microm. Explantation of the pIOL was refused by the patient. Seven years postoperatively, the ECD was 413 cells/mm(2) in the right eye and corneal decompensation occurred. The progressive unilateral endothelial loss was explained by excessive rubbing of the eyes because of chronic itching and an anterior shift of the pIOL over the 7 years as demonstrated by anterior optical coherence tomography. PMID- 19304105 TI - Spontaneous bilateral late-onset Descemet membrane detachment after successful cataract surgery. AB - We report the case of a 68-year-old man who developed bilateral Descemet membrane detachment (DMD) 4 weeks after successful cataract surgery and discuss the possible role of an underlying predisposition to DMD. Surgical intervention with gas injection in the anterior chamber resulted in excellent visual acuity restoration in the patient. To our knowledge, this is the first report of spontaneous bilateral DMD in the late postoperative period after cataract extraction. PMID- 19304106 TI - Anterior segment optical coherence tomography-aided diagnosis and primary posterior chamber intraocular lens implantation with fibrin glue in traumatic phacocele with scleral perforation. AB - We describe the case of a middle-aged woman who presented to us after injury from a clenched fist 3 days previously. The diagnosis was occult scleral perforation, severe conjunctival chemosis, and traumatic aphakia. However, the lens could not be localized during posterior segment examination. An anterior segment optical coherence tomography (AS-OCT) examination showed scleral discontinuity and a heterogeneous reflection in the subconjunctival area, suggesting a possible phacocele. Surgical exploration confirmed these findings. Aphakia was managed using the "glued intraocular lens" technique in the same sitting. This case highlights the use of AS-OCT in noncontact exploration of the traumatized anterior segment and in diagnosis of a possible phacocele along with an occult scleral perforation with uveal prolapse. To our knowledge, this is the first report of successful implantation of a glued IOL as a primary procedure combined with scleral perforation repair. PMID- 19304107 TI - Bilateral Salzmann-like nodular corneal degeneration after laser in situ keratomileusis imaged with anterior segment optical coherence tomography and high frequency ultrasound biomicroscopy. AB - Examination of a 34-year-old patient 7 years after laser in situ keratomileusis (LASIK) showed Salzmann-like nodules in both eyes, causing irregular astigmatism and decreased visual acuity. Two new imaging modalities were used to demonstrate the characteristics of this complication. Anterior segment optical coherence tomography (OCT) (Visante) scans showed hyperreflective nodules that varied in thickness from 0.35 mm to 0.56 mm. High-frequency ultrasound biomicroscopy illustrated the destruction of Bowman layer underlying these lesions. Both modalities demonstrated nodules as hyperreflective material covered by an abnormally thin layer of epithelium. Salzmann-like nodular degeneration is a rare but possible cause of decreased visual acuity and irregular astigmatism after LASIK surgery. Ultrasound biomicroscopy and OCT can demonstrate many of the established histopathologic findings in vivo. PMID- 19304108 TI - Acanthamoeba keratitis with perforation after corneal crosslinking and bandage contact lens use. AB - A 32-year-old man with keratoconus developed corneal melting 5 days after riboflavin/ultraviolet-A corneal collagen crosslinking (CXL). Corneal scraping was positive for Acanthamoeba. The patient was unaware that he was wearing a bandage contact lens and repeatedly rinsed his face and eyelids with tap water. Because of corneal perforation, a large therapeutic keratoplasty a chaud was performed. Although CXL is considered a safe procedure, this case emphasizes the potential risks. We discuss the potential effects of deepithelialization, contact lens placement, instillation of topical nonsteroidal antiinflammatory drugs and anesthetic agents, and the possible role of apoptosis when performing CXL treatment for keratoconus. PMID- 19304109 TI - Stiles-Crawford effect revisited: considerations for refractive surgery. PMID- 19304110 TI - Pilot study and functional results following implantation of the M-flex 630F multifocal intraocular lens. PMID- 19304111 TI - Scientific background on the risk engendered by reducing the lifetime blood donation deferral period for men who have sex with men. AB - The lifetime deferral for men who have sex with men (MSM) has not been harmonized with the 12-month deferral for similar-risk activities through heterosexual contacts. This occurs primarily because of fears of increased transfusion transmission of known sexually and transfusion-transmitted viruses for which donor blood is (eg, HIV) or is not (eg, human herpesvirus 8 [HHV-8]) tested and also of fears of novel agents that may share the epidemiology and long asymptomatic phase of HIV. A 12-month MSM deferral could result in release of 1 HIV-infectious donation every 11 years in the United States. This risk is smaller than the risk from allowing the continued use of pooled whole blood-derived platelets (release of 1 infectious platelet dose every 1.67 years), a risk that is considered "tolerable." Provided that measures to reduce the number of allogeneic-donor exposures to novel pathogens (which may be vector- or food-borne rather than sexually transmitted) are implemented, and the deferral for similar risk activities through heterosexual contacts is extended to 5 years, a 5-year MSM deferral could be justified because of the interval between emergence of a novel pathogen and introduction of measures to protect the blood supply. Also, provided that measures to protect the blood supply from HHV-8 are implemented, a lifetime MSM deferral could be justified because of the uncertainty about the clinical consequences of transfusion transmission of HHV-8. Because such alternate measures, which would have had a greater impact on safety than the MSM deferral, have not been implemented to demonstrate a consistent approach to safety, maintenance of the current MSM deferral appears to be selectively precautionary and cannot be justified. PMID- 19304112 TI - Blood iron homeostasis: newly discovered proteins and iron imbalance. AB - In biological systems, iron exerts 2 contrasting effects. The chemical reactivity of iron is essential for the biological activities of proteins such as hemoglobin, ribonucleotide reductase, the cytochromes, and aconitases. However, free iron in a cell has the propensity to generate free radicals which can damage cellular components containing proteins, lipids, and nucleic acids. To maintain the balance between iron as an essential nutrient and iron as a potential cytotoxin, a number of biological protective mechanisms have evolved. As shown in the thalassemias, iron imbalance can have devastating effects on human health. Recently, several new proteins have been described that play critical roles in iron regulation including the master regulator of iron metabolism (hepcidin). In this review, we discuss the new knowledge that has arisen from studies in yeast and in humans, and we show how these studies are shedding new light on some well known human disorders. PMID- 19304113 TI - Properties of pathogen-inactivated plasma components. AB - There are now 4 licensed technologies available for the pathogen inactivation of fresh frozen plasma in Europe. None of these are currently available in the United States, whereas in other geographic regions some are licensed others are not. This review addresses the different technologies available for the pathogen inactivation of plasma, their impact on the potency of the product, their efficacy in addressing microbiological contamination, as well as summarizing potential immunologic and toxicologic concerns. Published evidence of clinical efficacy is also reviewed as are various approaches to determining cost effectiveness. PMID- 19304114 TI - Passenger lymphocyte syndrome with or without immune hemolytic anemia in all Rh positive recipients of lungs from rhesus alloimmunized donors: three new cases and a review of the literature. AB - The passenger lymphocyte syndrome (PLS) is an unusual complication of solid organ transplantation, in which donor lymphocytes produce antibodies reactive with host red blood cell (RBC) antigens. Risks for PLS include highly lymphoid grafts, past sensitization of the donor against relevant RBC antigens, and donor lymphocyte escape of host immune clearance. For a 1-year period at our center, we observed an uncommonly high frequency of post-lung transplant Rhesus PLS, occurring once in every 31 cases. Passenger lymphocyte syndrome resulted from 2 alloimmunized cadaveric donors, in 3 of 3 D+, ABO-identical but HLA-unmatched recipients who initially had nonreactive RBC antibody screens. In case 1, the right lung of a group A, D-negative donor, with antibodies against D, C, and E antigens, was transplanted into a group A, R1R1 recipient. The recipient developed severe hemolytic anemia, direct antiglobulin test (DAT)-positive, on postoperative day (POD) 17. Anti-D and anti-C were identified on both the indirect antiglobulin test (IAT) and the RBC eluate. She required 10 U of RBCs in 40 days as well as plasmapheresis (POD 36-40). When transfusion dependence ceased, anti-D +/- C remained detectable on DAT and IAT for another 6.5 months. In case 2, the group A, R1r recipient of the same donor's left lung exhibited anti-D for the first time at posttransplant month 4 on both IAT and DAT. This activity persisted until a rejection episode 5 months later, without ever causing any evidence of hemolysis. In case 3, the group O, R1R1 recipient of both lungs of a group O, D negative donor, with antibodies against D, C, and V antigens, developed a nonhemolytic DAT and IAT with anti-D +/- C at postoperative month 2, which remained positive at last follow-up (6 months posttransplant). In conclusion, this report suggests a high incidence of Rhesus antibody PLS after lung transplantation, with wide variations in the timing of antibody onset, persistence, and severity. A review of the phenomenon and its implications are discussed. PMID- 19304115 TI - Rapid and accurate in vitro assays for detection of West Nile virus in blood and tissues. AB - West Nile virus (WNV) is a mosquito-borne single-stranded RNA virus, which has relatively recently emerged as a blood transfusion and organ transplantation transmissible pathogen. Low levels of WNV (viremia) are found in asymptomatic blood transfusion or cell/tissue donors with an infection, which poses a health threat to recipients. Since the introduction of nucleic acid testing (NAT) in 2003, many changes have occurred in the field of WNV detection and diagnosis. This review will focus on the recent progress in the in vitro assays for rapid and accurate detection of WNV in blood and tissues. PMID- 19304116 TI - Severe hemolytic anemia post-renal transplantation produced by donor anti-D passenger lymphocytes: case report and literature review. AB - The passenger lymphocyte syndrome (PLS), often associated with immune-mediated hemolytic anemia after solid organ and hematopoietic stem cell transplantation, is the result of concomitant transplantation of donor lymphocytes along with the donor allograft. Antibodies directed against recipient red blood cells (RBCs) are frequently found in ABO-mismatched solid organ transplants; however, passenger lymphocyte-mediated hemolysis due to Rh-incompatible antibodies has only rarely been reported. In this report, we present a case of severe hemolytic anemia related to the PLS in an ABO-matched renal allograft recipient. The recipient's blood type was A Rh(D) positive; and the donor, who had been previously alloimmunized, was A Rh(D) negative. The renal allograft recipient's hemoglobin abruptly decreased on postoperative day 12 in the setting of a newly positive direct antiglobulin test and anti-D antibodies in the plasma. The patient required intermittent RBC transfusions for ongoing hemolysis during the first 6 months post-renal transplant. Of all reported cases of anti-D-mediated PLS, our patient would seem to have been one of the most severe, as indicated by a nadir hemoglobin of 41 g/L and the need for 23 U of transfused RBCs. A hemolytic anemia occurring after organ transplantation should raise the possibility of donor derived antibodies directed against the recipient RBCs. Passenger lymphocyte syndrome-associated hemolysis is occasionally severe as in our case, but can be effectively treated with compatible RBC transfusions. PMID- 19304117 TI - Autologous adipose tissue as a new source of progenitor cells for therapeutic angiogenesis. AB - Therapeutic angiogenesis is an important means for salvaging tissues from severe ischemic diseases in patients with no option for other vascular interventions. A number of recent studies examined the possibilities of cell transplantation mediated angiogenesis using autologous bone marrow mononuclear cells, CD34(+) cells, peripheral mononuclear cells, etc. Subcutaneous adipose tissue can be harvested by relatively easy technology. Recent studies indicate that adipose tissue contains progenitor cells that can give rise to several mesenchymal lineages. Moreover, these progenitor cells can release multiple angiogenic growth factors including vascular endothelial growth factor, hepatocyte growth factor, and chemokine stromal cell-derived factor. The combination of these biological properties of adipose-derived regenerative cells indicates that autologous adipose tissue will be a useful cell source for therapeutic angiogenesis. PMID- 19304118 TI - Peak time of acute coronary syndrome in patients with sleep disordered breathing. AB - BACKGROUND: Recently, sleep disordered breathing (SDB) has gained attention in the field of cardiology. Until now, no study describing the relationship between acute coronary syndrome (ACS) and SDB has been carried out in Japan. METHODS: Among ACS patients admitted to our hospital, 44 patients (mean age 60.6+/-13.5 years) who received a portable polysomnography to measure apnea hypopnea index (AHI) were selected for this study. The circadian pattern of ACS onset was studied in 6-h intervals. In addition, all subjects were divided into three groups according to AHI severity (AHI < 5, 5 < or = AHI < 15, and 15 < or = AHI). Then, a comparative study between peak time of ACS and AHI severity was conducted for each group. RESULTS: In the AHI < 5 group, 66.0% patients suffered from ACS between 12:00 h and 18:00 h and 17.0% between 18:00 h and 24:00 h, and a total of 83.0% patients had ACS between 12:00 h and 24:00 h. In the 5 < or = AHI < 15 group, 49.9% patients had ACS between 24:00 h and 06:00 h, 16.7% patients between 06:00 h and 12:00 h. 12:00-18:00 h and 18:00-24:00 h showed no significant difference. All 22 patients in the 15 < or = AHI group suffered from ACS between 24:00 h and 12:00 h. CONCLUSION: The results of this study suggest a possible relationship between SDB and the onset of ACS between midnight to morning. PMID- 19304119 TI - Augmented sympathoinhibitory effect of valsartan when added to angiotensin converting enzyme inhibitor in patients with left ventricular dysfunction. AB - OBJECTIVE: Angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs) effectively interfere with the sympathetic nerve activity in patients with left ventricular (LV) dysfunction. The aim of this study was to examine the effect of ARBs on sympathetic nerve activity and baroreflex function in patients with LV dysfunction already receiving ACE inhibitors. METHODS: Twenty patients with LV dysfunction already treated with ACE inhibitor (enalapril 5 mg/day) were randomly divided into two groups: treatment with 10 mg/day enalapril (control group) or 5 mg/day enalapril plus 80 mg/day valsartan (combination group). In both groups, resting muscle sympathetic nerve activity (MSNA; microneurography), arterial baroreflex sensitivity, and cardiopulmonary baroreflex sensitivity were measured at baseline and 4 weeks after the treatment. Arterial baroreflexes were perturbed by phenylephrine method, and cardiopulmonary baroreflexes were perturbed by lower body negative pressure (-10 mmHg). RESULTS: Baseline characteristics in both groups were similar. Resting MSNA decreased significantly from 35.4+/-10.8 to 26.4+/-5.1 burst/min (p<0.05), while arterial baroreflex sensitivity improved significantly from 6.0+/-2.0 to 10.1+/-2.6 ms/mmHg in the combination group. Moreover, cardiopulmonary baroreflex control of MSNA improved significantly from 15.8+/ 12.2 to 42.0+/-26.7% (p<0.05) in the combination group. However, there were no significant changes in arterial baroreflex sensitivity and cardiopulmonary baroreflex of MSNA in the control group. CONCLUSION: Addition of ARB to ACE inhibitor treatment reduced sympathetic nerve activity and augmented arterial and cardiopulmonary baroreflex sensitivity in patients with LV dysfunction. PMID- 19304120 TI - Combination of conventional biomarkers for risk stratification in chronic heart failure. AB - BACKGROUND: Although there is substantial interest in the use of newer biomarkers to identify patients with chronic heart failure (CHF), recently few investigations have evaluated the incremental usefulness of multiple conventional biomarkers. Combination of several biomarkers simultaneously could enhance risk stratification in CHF. METHODS AND RESULTS: We analyzed 7 biomarkers (brain natriuretic peptide, uric acid, sodium, hemoglobin, creatinine, creatinine clearance, high-sensitivity C-reactive protein), which were known as established prognostic markers for CHF, in 154 consecutive CHF patients, and patients were prospectively followed with endpoints of cardiac death or re-hospitalization. When there was an abnormal value, we scored it for one point to calculate multimarker score. Patients were categorized into 3 strata according to multimarker score. There were 83 cardiac events during the follow-up period. A Cox proportional hazard model showed that patients in the high stratum were associated with the highest risk of cardiac events among the 3 strata. Kaplan Meier analysis revealed that patients in the high stratum had a significantly higher cardiac event rate compared with lower strata. CONCLUSION: The combination of conventional biomarkers could potentially improve the risk stratification of CHF patients for the prediction of cardiac events with low cost and wide availability. PMID- 19304121 TI - Accuracy of measuring mitral annular velocity by 2D speckle tracking imaging. AB - BACKGROUND: Recent developments in 2D speckle tracking imaging allow not only measurements of regional myocardial strain, but also velocities of the mitral annulus. The aim of this study was to determine the accuracy of speckle tracking derived mitral annulus velocity compared with conventional pulsed wave Doppler measurements. METHODS: 2D speckle tracking was acquired from the apical 4-chamber view (QLab, Speckle SQ, Philips, Andover, MA) in 169 subjects. While using texture tracking, two small regions of interest (ROIs) were placed in the septal (IVS) and lateral corners (LAT) of the mitral annulus. The software automatically tracked the ROIs frame-by-frame, yielding regional mean velocity curves of the mitral annulus throughout the cardiac cycle (synthetic pulsed wave Doppler; SPW). From these curves, peak systolic, early- and late-diastolic velocities of the mitral annulus (S'-SPW, E'-SPW, and A'-SPW) were measured. Peak systolic, early- and late-diastolic velocity in the mitral annulus (S', E', and A') by conventional pulsed wave tissue Doppler tracing were also obtained. RESULTS: Adequate ROI tracking was observed in 150/169 subjects in IVS and 139/169 subjects in LAT of the mitral annulus. All annular velocities derived from SPW were significantly lower than corresponding velocities obtained from conventional method in both IVS and LAT regions of the mitral annulus. However, significant correlation between S' (E', A')-SPW and S' (E', A') was observed. In particular, a good correlation between E'-SPW and E' was noted in both IVS (r=0.89, P<0.001) and LAT (r=0.85, P<0.001) regions of the mitral annulus. If we defined E/E'-SPW in IVS>26 for predicting E/E' in IVS>15, sensitivity, specificity, and accuracy were 83%, 97%, and 94%, respectively. E/E'-SPW in LAT>13 had a 94% sensitivity, 92% specificity, and 93% accuracy for predicting E/E' in LAT>10. CONCLUSIONS: The values of annular velocities by SPW were significantly lower compared to those assessed by traditional tissue velocities. However, new cut-off values of E/E' SPW for predicting elevated LV filling pressure highly correlated with traditional parameters. 2D speckle tracking imaging provides another strategy for evaluating LV filling pressures. PMID- 19304122 TI - Levels of serum deoxyribonuclease I activity on admission in patients with acute myocardial infarction can be useful in predicting left ventricular enlargement due to remodeling. AB - OBJECTIVES: Serum deoxyribonuclease I (DNase I) activity has recently been highlighted as a potential diagnostic marker for the early detection of an acute myocardial infarction (AMI). We evaluated whether the serum DNase I activity was associated with the parameters of the left ventricular (LV) remodeling after an AMI. METHODS: We measured the serum DNase I activity in 45 patients with an AMI who were admitted to our hospital within approximately 4 h of the onset of their chest pain. We also evaluated the LV ejection fraction (LVEF), LV end-diastolic volume (LVEDV), and LV end-systolic volume (LVESV) of each patient by echocardiography at the time of admission and at 6 months after the onset of the AMI. RESULTS: The serum DNase I activity peaked at 3.5+/-2.0 h after the onset of the symptoms in the patients with an AMI, thereafter exhibiting a time-dependent decline within 12 h, and a return to the basal level within 24 h in almost all cases. Neither the LVEF, LVEDV, nor LVESV in each patient on admission exhibited a significant correlation to the peak levels of the serum DNase I activity. Although there was no correlation between the peak DNase I activity and LVEF at 6 months after the onset, a significant positive correlation of the peak DNase I activity with LVEDV and LVESV (r=0.48, p<0.001 and r=0.34, p=0.02, respectively) was found. Furthermore, the LVEDV at 6 months after the onset in the high DNase I activity group (> 17.9 U/L) were significantly higher than those in the low DNase I activity group (< or = 17.9 U/L) (118.0+/-28.2 ml vs 89.3+/-25.4 ml, p=0.026). CONCLUSIONS: The serum DNase I activity level may predict LV enlargement associated with remodeling after an AMI. PMID- 19304123 TI - Increased mean platelet volume in rheumatic mitral stenosis: a possible factor for thromboembolic events. AB - BACKGROUNDS: Systemic embolism is an important complication in patients with rheumatic mitral stenosis (RMS). The mean platelet volume (MPV) is considered a marker and determinant of platelet function since larger platelets are hemostatically more reactive than platelets of normal size, increasing the propensity to thrombosis. The aim of this study was to investigate MPV in patients with RMS and healthy control subjects. METHODS: We selected 30 consecutive patients with RMS and 31 consecutive healthy age- and sex-matched control subjects. All subjects were in sinus rhythm. We measured MPV in a blood sample collected in EDTA. RESULTS: Demographic data of the RMS (mean age: 39.5+/ 9.9 years, 22 [71%] female) and control groups (mean age: 39.2+/-9.3 years, 20 [67%] female) were similar. The MPV was significantly higher in patients with RMS 8.8 (8.2-11.3) fl than control subjects 8.1 (7.1-9.3) (P<0.001). CONCLUSION: Elevated MPV might be considered as a marker of increased thromboembolic risk in patients with RMS. We suggest that patients with high MPV values might benefit from antiplatelet therapy. PMID- 19304124 TI - Coil occlusion of PDA in patients younger than 1 year: risk factors for adverse events. AB - BACKGROUND: Transcatheter occlusion of infantile patent ductus arteriosus (PDA) challenges the interventionist. PURPOSE: To analyze the risk factors for adverse events from this procedure in patients younger than 12 months. SUBJECTS: We retrospectively analyzed data on 32 patients younger than 12 months in whom transcatheter coil occlusion of a PDA was attempted. Ages ranged from 1 to 11 (median, 7) months and body weight from 1.2 to 10 (6.0) kg. The minimum ductal diameter ranged from 1.0 to 4.6 (3.3) mm and pulmonary to systemic flow ratio from 0.7 to 12.5 (2.2). Major adverse events were defined as those requiring surgery, while minor adverse events included transient hemolysis not needing treatment, coil migration with successful transcatheter retrieval, and mild left pulmonary artery (PA) stenosis. We determined whether any factors such as age, body weight, minimum PDA diameter, angiographic type, operator, and use of a 0.052-in. Gianturco coil related to the occurrence of adverse events. RESULTS: In two patients, coils could not be deployed in the appropriate position. They underwent surgery following transcatheter retrieval of coils. Coils were successfully deployed in the other 30 patients, however, one patient developed persistent hemolysis that required surgical retrieval of the coils and closure. PDA was completely closed in the other 29 patients (clinical success rate, 91%). Thus, there were three major adverse events, while minor adverse events occurred in five patients. Univariate analysis did not identify any single factor that contributed to either major, minor, or total adverse events. However, all major adverse events occurred in patients under 6 months and less than 6 kg body weight with a minimum duct diameter of more than 3.5 mm. CONCLUSION: Transcatheter coil occlusion of PDA is feasible in selected patients younger than 1 year. A minimum diameter more than 3.5 mm in patients under 6 kg may be a risk factor for major adverse events. PMID- 19304125 TI - Waon therapy improves the prognosis of patients with chronic heart failure. AB - BACKGROUND: We developed a Waon therapy (soothing warm therapy) and have previously reported that repeated Waon therapy improves hemodynamics, peripheral vascular function, arrhythmias, and clinical symptoms in patients with chronic heart failure (CHF). The aim of this study was to investigate the effect of Waon therapy on the prognosis of CHF patients. PATIENTS AND METHODS: We studied 129 patients with CHF in NYHA functional class III or IV who were admitted to our hospital between January 1999 and March 2001. In the Waon therapy group, 64 patients were treated with a far infrared-ray dry sauna at 60 degrees C for 15 min and then kept on bed rest with a blanket for 30 min. The patients were treated daily for 5 days during admission, and then at least twice a week after discharge. In the control group, 65 patients, matched for age, gender, and NYHA functional class, were treated with traditional CHF therapy. The follow-up time was scheduled for 5 years. RESULTS: Recent, complete follow-up data on each patient were obtained. The overall survival rate was 84.5% (Kaplan-Meier estimate). Twelve patients died in the control group and 8 patients died in the Waon therapy group at 60 months of follow-up. Cardiac events due to heart failure or cardiac death occurred in 68.7% of the control group but only 31.3% of the Waon therapy group (P<0.01) at 60 months of follow-up. CONCLUSION: Waon therapy reduced cardiac events in patients with CHF. This therapy is a promising non pharmacological treatment for CHF. PMID- 19304126 TI - Smoking cessation is associated with increased plasma adiponectin levels in men. AB - OBJECTIVES: Low levels of adiponectin, an adipocytokine with anti-diabetic and anti-atherogenic properties, are associated with increased risk of future myocardial infarction in men. Previous studies have demonstrated that cigarette smoking is involved in the development of insulin resistance, and current smokers have been shown to have reduced plasma adiponectin levels. However, the influence of smoking cessation on adiponectin levels remains unknown. We sought to assess whether smoking cessation is associated with increased plasma adiponectin levels in men. METHODS: The study includes 72 men (47 non-smokers and 25 current smokers at baseline) with stable angina pectoris who underwent percutaneous coronary intervention and follow-up coronary angiography 6 months later. During the 6 month follow-up period, all 47 non-smokers remained non-smokers, while 15 men of the 25 baseline current smokers successfully quit smoking. We evaluated plasma adiponectin levels at coronary intervention and 6 months later. RESULTS: Plasma adiponectin levels at coronary intervention were comparable to those after 6 months in non-smokers (4.22 [3.15-6.43] vs. 4.58 [3.03-6.26] microg/mL, P=0.124) and in persistent smokers (4.77 [4.25-10.53] vs. 5.16 [4.11-8.10] microg/mL, P=0.721). Meanwhile, an increase in adiponectin level was observed in patients who quit smoking for 6 months (4.24 [3.30-5.70] vs. 5.50 [4.03-8.00] microg/mL, P=0.002). Univariate analysis revealed that the percent increase in adiponectin levels correlated positively with smoking cessation (P=0.003) and negatively with additional use of beta-blockers (P=0.049). In addition, increases in adiponectin levels were closely associated with increase in high-density lipoprotein cholesterol (P=0.148), decrease in triglycerides (P=0.140), and additional use of renin-angiotensin system inhibitors (P=0.069). Multivariate analysis demonstrated that smoking cessation was an independent determinant of the increase in adiponectin (P=0.036). CONCLUSIONS: Smoking cessation is associated with increased plasma adiponectin levels in men with stable angina, suggesting that the significance of smoking cessation may be partly explained by the increase in adiponectin level. PMID- 19304127 TI - Endothelial dysfunction in a cohort of North Indian children with Kawasaki disease without overt coronary artery involvement. AB - OBJECTIVE: Kawasaki disease (KD) is a diffuse necrotizing vasculitis with predominant involvement of coronary arteries. Endothelial dysfunction has been implicated as an important event in the pathogenesis of arteriosclerosis, coronary vasoconstriction, hypertension, and myocardial ischemia. We examined the presence of endothelial dysfunction in North Indian children (of Caucasoid ethnicity) with KD without overt coronary artery involvement. METHODS: Twenty children (mean age 8.4+/-2.3 years; range 4.5-12.1 years) in the convalescent phase of KD were studied. All had received intravenous immunoglobulin during the acute phase of the disease. The interval between acute episode and enrolment ranged from 3 to 78 months (mean 25.3+/-20.1 months). High-resolution ultrasonography was used to analyze brachial artery responses to reactive hyperemia (with increased flow causing endothelium-dependent dilatation) and sublingual nitroglycerine (causing endothelium-independent dilatation). Flow mediated dilatation was also studied in an equal number of healthy age- and sex matched controls. Carotid artery stiffness index (SI) was calculated and compared in all subjects using previously published equations. RESULTS: Significant differences were observed between the percent flow mediated dilatation in children with KD (5.7+/-9.2%) compared with controls (12.2+/-8.9%, p=0.017). Sublingual nitroglycerine-mediated dilatation in children with KD was 28.5+/ 12.3%. Carotid artery SI was higher in children with KD (2.81+/-0.77 U) when compared to controls (2.32+/-0.80 U), but it failed to meet statistical significance (p=0.058). CONCLUSION: Brachial arteries of children with KD show evidence of endothelial dysfunction. This may point towards possibility of preclinical arteriosclerosis in children with KD even in the absence of coronary artery abnormalities. PMID- 19304128 TI - Estimated glomerular filtration rate as a predictor of secondary outcomes in Japanese patients with coronary artery disease. AB - BACKGROUND: Coronary artery disease (CAD) is highly prevalent and strongly associated with adverse outcomes in patients with chronic kidney disease (CKD). Recent data demonstrate that estimated glomerular filtration rate (eGFR) is more useful than serum creatinine as a predictor of outcomes. METHODS AND RESULTS: We investigated the clinical significance of eGFR-defined CKD in Japanese patients with CAD. In 702 consecutive patients with suspected CAD who underwent coronary angiography, CKD (eGFR <60 ml/min/1.73 m(2)) was present in 345 patients (49%). The eGFR value was lower in patients with multi-vessel coronary artery disease compared to patients with no significant stenosis (59+/-24 ml/min/1.73 m(2) vs 66+/-21 ml/min/1.73 m(2), p<0.01). During a follow-up period of 36 months, secondary events that included all-cause death and cardiovascular events requiring hospitalization occurred in 114 (16%) patients. Multivariate analysis using a Cox proportional hazards model showed that CKD [relative risk (RR) 1.707, 95%CI, 1.170-2.489; p=0.004] along with diabetes (RR, 1.684, 95%CI, 1.262-2.386; p=0.008) were independent predictors of secondary events. CONCLUSIONS: eGFR defined CKD is an important predictor of secondary outcomes in Japanese patients with CAD. Anti-atherosclerotic therapies under eGFR monitoring to consider renoprotection would be an important strategy to improve long-term prognosis in Japanese CAD patients. PMID- 19304129 TI - Abnormal Tei index predicts poor left ventricular mass regression and survival after AVR in aortic stenosis patients. AB - BACKGROUND: A Tei index is known to reflect overall cardiac performance including systolic and diastolic function in a variety of heart disease. We investigated the relationship between preoperative Tei index and postoperative left ventricular (LV) mass regression and survival after aortic valve replacement (AVR) for aortic valve stenosis (AS). METHODS: One hundred fifty-four patients with AS were classified into a group with abnormal (Abn) LV function (n=47, 0.45 < or = Tei index) and a group with normal (Nor) LV function (n=107, Tei index < 0.45). The pre- and postoperative echocardiographic variables including LV dimension, LV wall thickness, and LV mass regression as well as 6-year survival were compared between the two groups. RESULTS: There was a significant difference in both absolute and relative LV mass index (LVMI) regression (P=0.004 and 0.0007). Multiple linear regression analysis revealed that the preoperative LVMI, Tei index, and follow-up period were independent predictors of LVMI regression after AVR. Thirteen patients died (valve-related death in 5). Although the overall survival rate in the Nor-LV group (92.8%) was significantly better than that in the Abn-LV group (71.6%), there was no significant difference in survival free from valve-related death. CONCLUSIONS: Preoperative Tei index can be one of the significant predictors of LVMI regression and overall survival after AVR. PMID- 19304130 TI - Heart rate recovery after exercise in chronic heart failure: role of vital exhaustion and type D personality. AB - OBJECTIVE: Vital exhaustion and type D personality previously predicted mortality and cardiac events in patients with chronic heart failure (CHF). Reduced heart rate recovery (HRR) also predicts morbidity and mortality in CHF. We hypothesized that elevated levels of vital exhaustion and type D personality are both associated with decreased HRR. METHODS: Fifty-one patients with CHF (mean age 58+/-12 years, 82% men) and left ventricular ejection fraction (LVEF) < or = 40% underwent standard exercise testing before receiving outpatient cardiac rehabilitation. They completed the 9-item short form of the Maastricht Vital Exhaustion Questionnaire and the 14-item type D questionnaire asking about negative affectivity and social inhibition. HRR was calculated as the difference between heart rate at the end of exercise and 1 min after abrupt cessation of exercise (HRR-1). Regression analyses were adjusted for gender, age, LVEF, and maximum exercise capacity. RESULTS: Vital exhaustion explained 8.4% of the variance in continuous HRR-1 (p=0.045). For each point increase on the vital exhaustion score (range 0-18) there was a mean+/-SEM decrease of 0.54+/-0.26 bpm in HRR-1. Type D personality showed a trend toward statistical significance for being associated with lower levels of HRR-1 explaining 6.5% of the variance (p<0.08). The likelihood of having HRR-1 < or = 18 bpm was significantly higher in patients with type D personality than in those without (odds ratio=7.62, 95% CI 1.50-38.80). CONCLUSIONS: Elevated levels of vital exhaustion and type D personality were both independently associated with reduced HRR-1. The findings provide a hitherto not explored psychobiological explanation for poor cardiac outcome in patients with CHF. PMID- 19304131 TI - Comparison of determinations of left atrial volume by the biplane area-length and Simpson's methods using 64-slice computed tomography. AB - OBJECTIVES: There is increasing evidence that left atrial (LA) size is an important predictor of adverse cardiovascular outcomes such as atrial fibrillation, stroke, and congestive heart failure. The aim of this study was to determine whether there is a difference in results of quantification of LA volume by the area-length and Simpson's methods using multislice computed tomography (MSCT). METHODS AND RESULTS: The study population consisted of 51 patients with sinus rhythm (sinus group) and 20 patients with atrial fibrillation (af group) clinically indicated for MSCT angiography for evaluation of coronary arteries. Maximum LA volume, obtained at end-systole from the phase immediately preceding mitral valve opening, was measured using the area-length and Simpson's methods. In the sinus group, the mean LA volumes, indexed to body surface area, were 48.4+/-17.9 ml/m(2) with the area-length method and 48.3+/-17.0 ml/m(2) with the Simpson's method. In the af group, the mean indexed LA volumes with the area length method and the Simposon's method were 91.5+/-47.5 ml/m(2) and 90.3+/-45.9 ml/m(2), respectively. LA volumes calculated by the area-length method exhibited a strong linear relationship and agreement with those calculated using Simpson's method in both the groups (sinus group: r=0.99, P<0.0001, af group: r=0.99, P<0.0001). CONCLUSIONS: The area-length method is a simple and reproducible means of assessment of LA volume. Standardization of LA volume assessment using MSCT is important for serial follow-up and meaningful communication of results of testing among institutions and physicians. PMID- 19304132 TI - Practical assessment of myocardial viability with a positron coincidence gamma camera using 18F-fluorodeoxyglucose in acute myocardial infarction: comparison with dedicated positron emission tomography and 201Tl single photon emission computed tomography. AB - OBJECTIVE: 2-[(18)F] fluoro-2-deoxy-D-glucose ((18)F-FDG) began to be supplied commercially to our hospital, which does not have a cyclotron, in autumn of 2005. The purpose of this study was to compare the utility of a dual-head positron coincidence detection (PCD) gamma camera in the detection of myocardial viability using (18)F-FDG with that of dedicated positron emission tomography (PET) and with that of thallium-201 ((201)Tl) single photon emission computed tomography (SPECT). METHODS: A total of 15 patients (14 men and 1 woman, mean age: 60+/-7 years, range: 46-73) with a large acute myocardial infarction (AMI) underwent (18)F-FDG PET, (18)F-FDG PCD imaging after oral glucose loading (75 g) and (201)Tl SPECT imaging. We divided the SPECT and PET images into a total of 20 segments, and semiquantitative visual analysis was performed by assessing regional tracer activities on a 4-point scoring system (DS): 0=normal uptake, 1=mildly reduced uptake, 2=severely reduced uptake, and 3=no uptake. We summed the DS in each patient as the total DS (TDS). RESULTS: The TDS of the (18)F-FDG PET image was 14.4+/-7.7. The TDS of the (18)F-FDG PCD image was 18.7+/-7.7. The TDS of the (201)Tl SPECT image was 24.1+/-11.5. The TDS of the (18)F-FDG PET image was significantly smaller than that of the (18)F-FDG PCD image. The TDS of the (18)F-FDG PET image was significantly smaller than that of the (201)Tl SPECT image. The TDS of the (18)F-FDG PCD image was significantly smaller than that of the (201)Tl SPECT image. CONCLUSION: The findings of the project suggest that (18)F-FDG PCD is a good modality based on its accuracy, convenience, and cost performance for detecting myocardial viability in hospitals that do not have a PET system. PMID- 19304133 TI - Impact of acute hyperglycemia during primary stent implantation in patients with ST-elevation myocardial infarction. AB - BACKGROUND: Acute hyperglycemia is associated with increased mortality rates in patients with acute coronary syndrome. OBJECTIVE: This study aimed to evaluate the relationship between the glucose level and clinical variables during primary intervention in patients with ST-elevation acute myocardial infarction (STEMI). METHODS AND RESULTS: Of consecutive 94 patients with STEMI treated by primary stent implantation, acute hyperglycemia (plasma glucose level on admission>198 mg/dl) was recognized in 29 patients. There were no significant differences in baseline characteristics, except for the presence of diabetes and HbA(1c) level, between patients with and without acute hyperglycemia. In patients with acute hyperglycemia, corrected TIMI frame counts were significantly higher compared with those in patients without acute hyperglycemia (46.3+/-30.3 vs. 34.0+/-17.9, p=0.02). And corrected TIMI frame count was independently associated with plasma glucose level (p=0.006). Maximum level of creatine kinase (CK) and CK-MB were significantly higher in patients with acute hyperglycemia (CK, 4840.0+/-4690.3 vs. 2410.7+/-2302.9 IU, p=0.001; CK-MB, 315.3+/-257.7 vs. 195.9+/-191.1, p=0.01). CONCLUSION: The presence of acute hyperglycemia was associated with the impairment of epicardial coronary flow after primary stent implantation. This mechanism might be responsible for the increased infarct size. PMID- 19304134 TI - Positive remodeling is associated with more plaque vulnerability and higher frequency of plaque prolapse accompanied with post-procedural cardiac enzyme elevation compared with intermediate/negative remodeling in patients with acute myocardial infarction. AB - BACKGROUND: We assessed the impact of remodeling patterns on pre- and post procedural intravascular ultrasound (IVUS) findings and cardiac enzyme elevation after stenting in 310 acute myocardial infarction (AMI) patients. METHODS: The positive remodeling (PR) (PR group, n=113) was defined as remodeling index (lesion/reference external elastic membrane cross-sectional area) >1.05, intermediate remodeling (IR) as between 0.95 and 1.05, and negative remodeling (NR) as<0.95 (IR/NR group, n=197). IVUS findings included ruptured plaque (a cavity that communicated with the lumen with an overlying residual fibrous cap fragment), multiple ruptured plaques (different plaque ruptures separated by a >5 mm length of artery containing smooth lumen contours), thrombus (discrete intraluminal filling defects), and plaque prolapse (tissue extrusion through the stent strut at post-stenting). We compared pre- and post-procedural IVUS findings and cardiac-specific troponin I (cTnI) elevation after stenting according to the remodeling pattern. RESULTS: The plaque rupture (60% vs. 42%, p=0.004), multiple plaque ruptures (22% vs. 14%, p=0.014), and IVUS-detected thrombus (42% vs. 28%, p=0.012) were more common in the PR group compared with the IR/NR group. Post stenting plaque prolapse was observed more frequently (36% vs. 22%, p=0.008), and cTnI was elevated more significantly after stenting in the PR group compared with the IR/NR group (DeltacTnI; +7.8+/-51.1 ng/ml vs. +0.9+/-41.1 ng/ml, p=0.008). Multivariate analysis showed that PR [odds ratio (OR)=1.92; 95% CI 1.04-2.98, p=0.028], plaque rupture (OR 1.98; 95% CI 1.16-3.45, p=0.025), IVUS-detected thrombus (OR 2.30; 95% CI 1.22-3.98, p=0.008), and plaque prolapse (OR 8.40; 95% CI 4.19-16.84, p<0.001) were independently associated with post-stenting cTnI elevation. CONCLUSIONS: AMI patients with PR have more plaque vulnerability and higher frequency of plaque prolapse accompanied by post-procedural cardiac enzyme elevation compared with AMI patients with IR/NR. PMID- 19304135 TI - Coronary arterial spasm during adenosine myocardial perfusion imaging. AB - Adenosine is widely used as a pharmacologic agent for stress myocardial perfusion imaging. Vasospasm as a side effect of adenosine has been reported a few times in other countries, but it has not been reported in Japan. A 65-year-old woman was admitted to our hospital because of chest pain at rest and during exercise. She underwent myocardial scintigraphy, to rule out myocardial ischemia. After adenosine infusion, she felt chest pain and the electrocardiogram (ECG) showed ST elevation in inferior leads. Adenosine infusion was stopped immediately. Her chest pain resolved, and the ECG reverted to baseline. Perfusion image presented reverse redistribution in inferior segments, and coronary angiography revealed insignificant lesions. Transient ST elevation during adenosine infusion is thought to be due to coronary vasospasm, judging from scintigraphic and angiographic findings. PMID- 19304136 TI - A case of giant coronary artery aneurysm and literature review. AB - A 40-year-old man was referred to our hospital because of an abnormal shadow on the left cardiac border on the chest roentgenogram at the regular medical health examination without any symptoms. A giant coronary artery aneurysm of left anterior descending artery with a maximum diameter of approximately 50 mm was detected with computed tomography and coronary angiography. The patient was treated and followed up medically. Four years later, the size of the coronary artery aneurysm became larger. Then resection of the coronary artery aneurysm and coronary artery bypass grafting were successfully performed. Coronary artery aneurysms are rare in adults and are usually found in association with Kawasaki disease, coronary atherosclerosis, and so on. We also review the literature of giant coronary artery aneurysms exceeding 50 mm in diameter. PMID- 19304137 TI - Experience of appendicular thermal therapy applied to a patient with a left ventricular assist device awaiting heart transplantation. AB - Thermal therapy for heart failure is recognized to improve clinical symptoms. We describe our experience with appendicular thermal therapy applied to a patient fitted with an extracorporeal left ventricular assist device (LVAD) who was wait listed for a heart transplant. A 21-year-old male with end-stage heart failure due to dilated cardiomyopathy was fitted with a LVAD. His general condition stabilized after LVAD placement and the status of his heart failure has remained at NYHA class II for the past 13 months. However, his cardiac function did not sufficiently recover to discontinue LVAD support. We conducted appendicular thermal therapy using a steam foot bath and heated gloves for 2 weeks. Immediately after thermal therapy, his average sublingual temperature increased from 36.3 to 37.0 degrees C and the grade of mitral regurgitation, as well as LV ejection fraction and endothelial function improved. Furthermore, levels of oxidative and anti-oxidative stress markers decreased and increased, respectively, after 2 weeks of therapy. No complications developed. We conclude that appendicular thermal therapy was safe in this patient waiting for a heart transplant and who had an extracorporeal LVAD, and that the procedure might be beneficial for others with end-stage heart failure. PMID- 19304138 TI - Takotsubo cardiomyopathy associated with autoimmune polyendocrine syndrome II. AB - Takotsubo cardiomyopathy (TCM) is a poorly understood condition in which patients with chest pain have a transient ampulla-shaped abnormality of the left ventriculogram, and intact coronary arteries. We report TCM in combination with autoimmune polyendocrine syndrome type II (APS II), which raises new questions about the pathogenesis of TCM. PMID- 19304139 TI - Novel neointimal formation over sirolimus-eluting stents identified by coronary angioscopy and optical coherence tomography. AB - Neointimal proliferation after sirolimus-eluting stent (SES) implantation is generally inhibited by the pharmacological effects of sirolimus in comparison to bare metal stent (BMS). Neointimal hyperplasia after BMS implantation is mainly composed of vascular smooth muscle cells, and is usually observed as a white mass by angioscopy and as a layer of uniform signal intensity without attenuation on optical coherence tomography (OCT). In this case, angioscopic color of the neointima covering the SES was obviously yellow and OCT signal patterns of the neointima showed rapid attenuation similar to lipid tissues in atherosclerotic lesions. These findings suggest that neointima within the SES is quite different from that of the BMS and may contain atherosclerotic components. PMID- 19304140 TI - Delayed gadolinium enhancement and elevated plasma brain natriuretic peptide are useful in differentiating hypertrophic cardiomyopathy from athlete's heart. PMID- 19304142 TI - Abstracts of the 2009 European College of Neuropsychopharmacology (ECNP) Workshop on Neuropsychopharmacology for Young Scientists in Europe. March 5-8, 2009. Nice, France. PMID- 19304250 TI - Nutrition educators--providing practical support for families. PMID- 19304251 TI - Our invisible pillars. PMID- 19304252 TI - Are family meal patterns associated with overall diet quality during the transition from early to middle adolescence? AB - OBJECTIVE: To examine longitudinal associations of participation in regular family meals (>or= 5 meals/week) with eating habits and dietary intake during adolescence. DESIGN: Population-based, longitudinal study (Project EAT: Eating Among Teens). Surveys were completed in Minnesota classrooms at Time 1 (1998 1999) and by mail at Time 2 (2003-2004). SETTING: Baseline surveys were completed in Minneapolis/St. Paul, Minnesota, schools and by mail at follow-up. PARTICIPANTS: 677 adolescents (303 males and 374 females) who were in middle school at Time 1 (mean age = 12.8 +/- 0.74 years) and high school at Time 2 (mean age = 17.2 +/- 0.59 years). MAIN OUTCOME MEASURES: Dietary intake, frequency of meals, and fast-food intake patterns. ANALYSIS: Generalized linear modeling stratified by gender and adjusted for race/ethnicity, socioeconomic status, and the Time 1 outcome. RESULTS: Regular family meals were positively associated with Time 2 frequency of breakfast, lunch, and dinner meals for males and breakfast and dinner meals for females. Among males, regular family meals were negatively associated with Time 2 fast-food intake. Regular family meals were also positively associated with Time 2 mean daily intakes of vegetables, calcium-rich food, fiber, calcium, magnesium, potassium, iron, zinc, folate, and vitamins A and B(6) among both genders. CONCLUSIONS AND IMPLICATIONS: Regular family meals during early adolescence may contribute to the formation of healthful eating habits 5 years later. Parents should be made aware of the importance of shared mealtime experiences. PMID- 19304253 TI - Low-income, African American adolescent mothers and their toddlers exhibit similar dietary variety patterns. AB - OBJECTIVE: To examine the relationship between maternal and toddler dietary variety. DESIGN: Longitudinal; maternal and toddler dietary data were collected at 13 months; anthropometry was collected at 13 and 24 months. SETTING: Data were collected in homes. PARTICIPANTS: 109 primiparous, low-income, African American adolescent mothers and toddlers. MAIN OUTCOME MEASURES: Maternal and toddler dietary variety and toddler obesity at 24 months. ANALYSIS: Correlations were computed to estimate associations between maternal and toddler dietary variety at 13 months; multiple logistic regression analyses were conducted to estimate associations between maternal and toddler diet and toddler growth. RESULTS: Maternal and toddler fruit, vegetable, snack, meat, dairy, and soda variety were significantly correlated. There was no association between maternal and toddler dietary variety and obesity at 24 months. Adolescent mothers who purchased groceries consumed more fruits and vegetables and provided more variety for their toddlers than those who relied on others to purchase groceries. CONCLUSIONS AND IMPLICATIONS: Adolescent mothers and toddlers exhibited similar dietary patterns; consuming more sweets and less fruits and vegetables than recommended. Toddlerhood is an optimal time to address healthful dietary patterns and to help adolescent mothers influence grocery purchasing decisions. Goals are to establish healthful dietary patterns and reduce pediatric obesity. PMID- 19304254 TI - Diet- and body size-related attitudes and behaviors associated with vitamin supplement use in a representative sample of fourth-grade students in Texas. AB - OBJECTIVE: To examine diet- and body size-related attitudes and behaviors associated with supplement use in a representative sample of fourth-grade students in Texas. DESIGN: Cross-sectional data from the School Physical Activity and Nutrition study, a probability-based sample of schoolchildren. Children completed a questionnaire that assessed supplement use, food choices, diet related attitudes, and physical activity; height and weight were measured. SETTING: School classrooms. PARTICIPANTS: Representative sample of fourth-grade students in Texas (n = 5967; mean age = 9.7 years standard error of the mean [SEM] = .03 years, 46% Hispanic, 11% African-American). MAIN OUTCOME MEASURES: Previous day vitamin supplement consumption, diet- and body size-related attitudes, food choices, demographic factors, and physical activity. ANALYSIS: Multivariable logistic regression models, P < .05. RESULTS: The prevalence of supplement use was 29%. Supplement intake was associated with physical activity. Girls who used supplements were more likely to report positive body image and greater interest in trying new food. Relative to nonusers, supplement users were less likely to perceive that they always ate healthful food, although supplement use was associated with more healthful food choices in boys and girls (P < .001). CONCLUSIONS AND IMPLICATIONS: The widespread use of supplements and clustering of supplement use with healthful diet and greater physical activity in fourth graders suggest that supplement use be closely investigated in studies of diet disease precursor relations and lifestyle factors in children. PMID- 19304255 TI - Effect of nutrition intervention using a general nutrition course for promoting fruit and vegetable consumption among college students. AB - OBJECTIVE: To evaluate the effectiveness of implementing nutrition intervention using a general nutrition class to promote consumption of fruits and vegetables in college students. DESIGN: 3-day food records were collected, verified, and analyzed before and after the intervention. SETTING: A midwestern university. PARTICIPANTS: 80 college students, ages 18 to 24, participated in the study. INTERVENTION: The intervention focused on nutrition knowledge related to prevention of chronic diseases, healthful dietary choices increasing fruit and vegetable consumption, dietary feedback, and interactive hands-on activities. MAIN OUTCOME MEASURES: Consumption of: total vegetable, fresh vegetable, starchy vegetable, french fries, vegetable juice, total fruit, fresh fruit, canned fruit, and fruit juice. ANALYSIS: Dependent t test was used to analyze the differences in pre- and posttest. Analysis of variance was used to determine differences in dietary changes between groups. RESULTS: Participants significantly increased consumption of not only total fruits and vegetables (P < .005), but also fresh fruits and vegetables (P < .005). Intake of french fries decreased significantly (P < .05). Females responded better to the intervention than males in increasing vegetable consumption (P < .05). CONCLUSIONS AND IMPLICATIONS: Class-based nutrition intervention focusing on prevention of chronic diseases is a cost effective approach to increasing fruit and vegetable consumption among college students. PMID- 19304256 TI - Assessment of stage of change, decisional balance, self-efficacy, and use of processes of change of low-income parents for increasing servings of fruits and vegetables to preschool-aged children. AB - OBJECTIVE: Use the Transtheoretical Model of Behavior Change (TTM) to determine the proportionate stage of change of low-income parents and primary caregivers (PPC) for increasing accessibility, measured as servings served, of fruits and vegetables (FV) to their preschool-aged children and evaluate response differences for theoretical constructs. DESIGN: Cross-sectional, quantitative survey design consisting of staging algorithm, construct scales, and food frequency questionnaire. SETTING: Rural and urban communities in a southwestern state of the United States. PARTICIPANTS: 238 low-income PPC enrolled in federal nutrition education programs were recruited from group nutrition education sessions. MAIN OUTCOME MEASURES: Stage of change using a staging algorithm, TTM constructs of processes of change, decisional balance, and self-efficacy measured by multiple-item scales using Likert response, and fruit and vegetable servings served using a food frequency questionnaire. ANALYSIS: Descriptive analysis, Pearson's chi-square, analyses of variance with Tukey's Honestly Significant Difference post hoc test, and principal component function analysis. RESULTS: Of the surveyed PPC, 43% were in precontemplation/contemplation stages, and 29% were in the preparation stage for increasing FV accessibility (measured by servings served) to their preschool-aged children. PPC in the action/maintenance stages evidenced greater use of behavioral processes and had higher self-efficacy scores compared to PPC in precontemplation/contemplation and preparation stages. CONCLUSIONS AND IMPLICATIONS: Interventions aimed at increasing FV accessibility for preschool-aged children should be tailored to meet PPCs' stage of change. Interventions targeting PPC in precontemplation/contemplation stages should use methods to share ideas for planning meals and snacks to include FV. Interventions for PPC in the preparation stage should aim to build skills in quick preparation of economical FV, address parental role modeling of FV consumption, and encourage goal setting. Learning formats providing social support may prove effective in prevention of behavior relapse for PPC in action/maintenance stages. PMID- 19304257 TI - Applying precede-proceed to develop an intuitive eating nondieting approach to weight management pilot program. AB - OBJECTIVE: To describe the use of a consolidated version of the PRECEDE-PROCEED participatory program planning model to collaboratively design an intuitive eating program with Fort Drum military spouses tailored to their readiness to reject the dieting mentality and make healthful lifestyle modifications. DESIGN: A consolidated version of PRECEDE-PROCEED guided demographic, epidemiological, behavioral, organizational, and administrative diagnosis through survey research. Focus groups composed of planning/steering committee members diagnosed environmental, organizational, administrative. and policy considerations. Objectives were set for each phase to assist with program tailoring. SETTING: Recruitment at Fort Drum Army Installation, NY, summer 2004. PARTICIPANTS: Ninety one military health beneficiaries aged 20-65 years of age completed the pilot tested survey packet. PHENOMENON OF INTEREST: The survey assessed quality of life issues, Diet Mentality, Healthy Eating Index, and Intuitive Eating Stages of Change scores, and desired program mix. ANALYSIS: Mean and mode of survey responses and scores. RESULTS: A 10-week "My Body Knows When" intuitive eating program was tailored to increase attendance, reduce barriers, and increase successful rejection of a dieting mentality. CONCLUSIONS AND IMPLICATIONS: A consolidated version of PRECEDE-PROCEED efficiently guided participatory planning to tailor this program. PMID- 19304258 TI - Pilot study: EatFit impacts sixth graders' academic performance on achievement of mathematics and english education standards. AB - OBJECTIVE: Investigate the impact of a nutrition education program on student academic performance as measured by achievement of education standards. DESIGN: Quasi-experimental crossover-controlled study. SETTING: California Central Valley suburban elementary school (58% qualified for free or reduced-priced lunch). PARTICIPANTS: All sixth-grade students (n = 84) in the elementary school clustered in 3 classrooms. INTERVENTION: 9-lesson intervention with an emphasis on guided goal setting and driven by the Social Cognitive Theory. MAIN OUTCOME MEASURE: Multiple-choice survey assessing 5 education standards for sixth-grade mathematics and English at 3 time points: baseline (T1), 5 weeks (T2), and 10 weeks (T3). ANALYSIS: Repeated measures, paired t test, and analysis of covariance. RESULTS: Changes in total scores were statistically different (P < .05), with treatment scores (T3 - T2) generating more gains. The change scores for 1 English (P < .01) and 2 mathematics standards (P < .05; P < .001) were statistically greater for the treatment period (T3 - T2) compared to the control period (T2 - T1). CONCLUSION AND IMPLICATIONS: Using standardized tests, results of this pilot study suggest that EatFit can improve academic performance measured by achievement of specific mathematics and English education standards. Nutrition educators can show school administrators and wellness committee members that this program can positively impact academic performance, concomitant to its primary objective of promoting healthful eating and physical activity. PMID- 19304259 TI - Comparison of fast-food and non-fast-food children's menu items. AB - OBJECTIVE: Compare the macronutrient content of children's meals sold by fast food restaurants (FFR) and non-fast-food restaurants (NFF). DESIGN: All restaurants within the designated city limits were surveyed. Non-fast-food children's meals were purchased, weighed, and analyzed using nutrition software. All fast-food children's meals were recorded, with nutrient content information obtained from their respective commercial Web sites. SETTING: Community in southeastern United States. PARTICIPANTS: 33 restaurants, 10 FFR and 23 NFF. MAIN OUTCOME MEASURE(S): Total weight, total calories, fat, saturated fat, protein, carbohydrates, and fiber. ANALYSIS: Paired t tests, 1-way analysis of variance, and general linear model analyses. RESULTS: Overall mean percentage of calories from fat for children's meals at FFR was 37.5%, and 40.5% at NFF. Significant differences (P < .01) existed between FFR and NFF, with FFR providing smaller servings, fewer calories, and less total fat, saturated fat, protein, carbohydrates, and fiber. When analyzing only with the highest calorie option side, controlling for portion size, no differences remained except saturated fat. CONCLUSIONS AND IMPLICATIONS: Fast-food restaurants offered smaller children's portion sizes and lower-fat options. These results have implications for restaurants, parents, children, and nutrition educators. PMID- 19304260 TI - Development of core competencies for paraprofessional nutrition educators who deliver food stamp nutrition education. AB - The purpose of this project was to describe the process used for the development of core competencies for paraprofessional nutrition educators in Food Stamp Nutrition Education (FSNE). The development process included the efforts of an expert panel of state and multicounty FSNE leaders to draft the core competencies and the validation of those competencies by FSNE paraprofessionals. The result of the project was a comprehensive list of 10 core competency areas with specific competencies for each. The core competencies will be useful with FSNE for state and local program planning, implementation, evaluation, and decision making, with possible implications for other community-based education programs. PMID- 19304261 TI - Teaching the "health at every size" paradigm benefits future fitness and health professionals. PMID- 19304262 TI - Cultural experience project: expanding college students' worldview. PMID- 19304263 TI - [Strong genetic differentiation of the gilthead sea bream Sparus aurata (L., 1758) between the two western banks of the Mediterranean]. AB - We characterised four samples of gilthead sea bream from the two western Mediterranean banks with three microsatellite loci and two RAPDs systems. Contrarily to what could be predicted for a highly mobile species with a planktonic larval dispersal phase, we observed a strong and significant genetic differentiation at all loci between the two banks (F(st)=0.069(***)), whereas two samples from the Gulf of Lions were almost identical (F(st)=0.003 ns) while the two from the Gulf of Annaba displayed varied levels of differentiation according to the molecular marker considered. RAPDs showed a similar trend as microsatellites. The reasons for this surprisingly strong genetic differentiation, as compared to what has been observed in other species over comparable geographical distance, may be sought for either in a smaller as suspected larval dispersal, or in the non-neutrality of the loci studied. PMID- 19304264 TI - ISSCOR: Intragenic, Stochastic Synonymous Codon Occurrence Replacement--a new method for an alignment-free genome sequence analysis. AB - Synonymous codons do not occur at equal frequencies. Codon usage and codon bias have been extensively studied. However, the sequential order in which synonymous codons appear within a gene has not been studied until now. Here we describe an in silico method, which is the first attempt to tackle this problem: to what extent this sequential order is unique, and to what extent the succession of synonymous codons is important. This method, which we called Intragenic, Stochastic Synonymous Codon Occurrence Replacement (ISSCOR), generates, by a Monte Carlo approach, a set of genes which code for the same amino acid sequence, and display the same codon usage, but have random permutations of the synonymous codons, and therefore different sequential codon orders from the original gene. We analyze the complete genome of the bacterium Helicobacter pylori (containing 1574 protein coding genes), and show by various, alignment-free computational methods (e.g., frequency distribution of codon-pairs, as well as that of nucleotide bigrams in codon-pairs), that: (i) not only the succession of adjacent synonymous codons is far from random, but also, which is totally unexpected, the occurrences of non-adjacent synonymous codon-pairs are highly constrained, at strikingly long distances of dozens of nucleotides; (ii) the statistical deviations from the random synonymous codon order are overwhelming; and (iii) the pattern of nucleotide bigrams in codon-pairs can be used in a novel way for characterizing and comparing genes and genomes. Our results demonstrate that the sequential order of synonymous codons within a gene must be under a strong selective pressure, which is superimposed on the classical codon usage. This new dimension can be measured by the ISSCOR method, which is simple, robust, and should be useful for comparative and functional genomics. PMID- 19304265 TI - The dynamic changing of Ca2+ cellular localization in maize leaflets under drought stress. AB - Maize cultivar zhengdan958 was selected as materials. The sub-cellular distribution of soluble calcium at different phases was shown by the potassium pyroantinonate-precipitation method and transmission electron microscopy. The results showed that the deposits of calcium antimonate as the indicator for Ca(2+) localization were mainly concentrated within the vacuoles and intercellular spaces without PEG treatment. Firstly, when the leaf was treated with PEG, the Ca(2+) level increased remarkably in the cytoplasm, but considerably decreased in vacuoles and intercellular gaps. Meanwhile, the level of Ca(2+) also increased in chloroplast and nucleus. When the treatment continued, the level of Ca(2+) in chloroplasts and nucleus continued to increase and some cells and chloroplasts finally disintegrated, showing that there is a relationship between the distribution of Ca(2+) and the super-microstructure of cells. Ca(2+) plays a role in the plant drought resistance. The changes of cytosolic Ca(2+) localization in cells treated by ABA, EGTA, Verapamil and TFP were investigated too. The increase of cytosolic calcium induced by ABA was mainly caused by calcium influx. Calmodulin participated in ABA signal transduction, which was indicated by the variation of cytosolic Ca(2+)/CaM concentration change induced by ABA. The above results provided a direct evidence for calcium ion as an important signal at the experimental cellular level. PMID- 19304266 TI - Physiological behaviour of four rapeseed cultivar (Brassica napus L.) submitted to metal stress. AB - Eliminating heavy metals in the environment by phytoremediation is a method that uses, generally, plants with a low biomass yielded and feeble depth of root system. For the purpose of improving this technique, we have tested four varieties of productive specie with high yields, the rapeseed (Brassica napus L.). In particular, we have studied metal stress effect on biomass, growth, and endogenous Zn and Cd content. Metal treatment caused significant dry weight differences between metal-treated and control plants. A significant genotypic difference has been noticed between the four cv. For two varieties, Jumbo and Drakkar, the accumulation is more important in the stems and petioles, whereas, this accumulation is at a maximum level in the root system for the two varieties, Cossair and Pactol. Chlorophyll and carotenoid content, as well as lipid peroxidation, known as stress markers, were also evaluated. Metal treatment led to an increase in the amount of malondialdehyde (MDA) in the leaves. However, the increase of Zn and Cd levels in the tissue culture was followed by a decrease in the photosynthetic pigments. PMID- 19304267 TI - [Analgesic activity of the aqueous extract from Ximenia Americana]. AB - Pharmacological studies were conducted with the aqueous extract of the bark of the stem of Ximenia americana Linne (Olacaceae) on experimental animals, evaluating the analgesic activities. In the analgesic test, the aqueous extract elicited an inhibitory intensity on the acetic acid-induced writhing response and on the late phase of the formalin test, but possessed only a weak effect on the tail-flick response and on the early phase of the formalin test. PMID- 19304268 TI - [Activities in retired people and the risk of dementia]. AB - It is necessary to develop the prevention of Alzheimer's disease, because of the increase in the number of cases and unavailability of a curative treatment. From the data of the cohort PAQUID, we studied the risk of dementia according to leisure activities and the age of cessation of professional activity. The practice of a sport and reading decreases by 25% the risk of dementia during 15 years. The age of cessation of professional activity is not associated with the risk of dementia. An active life seems to be a possible way to prevent dementia. PMID- 19304269 TI - Study of nitrate leaching and nitrogen fate under intensive vegetable production pattern in northern China. AB - Because of intensive vegetable production in plastic greenhouses in northern China, the potential risk of nitrate leaching to groundwater is increasingly apparent, threatening ecosystem services and the sustainability of food production. In the present work, nine drainable lysimeters were installed into vegetable fields, with in-situ loamy soils, in Shouguang City of the north China vegetable base. The experiments were conducted to quantify the magnitude and variability of nitrate leaching to groundwater and to access the fate of total fertilizer-N inputs in the area. The results obtained indicated that: under local conventional agronomic practices, there is a high discrepancy in leaching nitrate N concentration (ranging from 17 to 457 mg L(-1)), and nitrate losses (152-347 kg N ha(-1)) were observed from 1-m soil profiles in the field. Meanwhile, high fertilizer N application resulted in low N efficiency, with only (33.0+/-13)% (mean+/-S.D.) of input N absorbed by the crops, while additionally nearly half of the total inputs of N were unaccounted in a partial N balance sheet. It is concluded that groundwater pollution associated with greenhouse-based vegetable production had been confirmed in Shouguang, adversely affecting water quality and leading to serial agro-ecological problems. PMID- 19304270 TI - Genetic diversity analysis of wild close relatives of barley from Tibet and the Middle East by ISSR and SSR markers. AB - The genetic diversity analysis of 90 barley samples, including 45 wild close relatives of barley from the Tibet region of China and 45 wild accessions from different countries throughout the Middle East, were carried out using ISSR and SSR markers. The results showed that Tibetan wild close relatives of barley had a higher genetic diversity than those from the Middle East. Ten ISSR primers amplified 91 allelic variants, of which 79 were polymorphic (86.81%), in the Tibetan genotypes and 82 allelic variants, of which 66 were polymorphic (80.49%), in the Middle East genotypes. Eleven primer pairs of SSR markers amplified 100 allelic variants among the Tibetan genotypes with 100 polymorphic bands (100%). Among the Middle East genotypes, 78 allelic variants were produced containing 77 polymorphic bands (98.72%). Moreover, the total gene diversity analysis (HT) values of Tibetan barley (0.227 for ISSRs and 0.126 for SSRs) were higher than those of Middle East (0.212 for ISSRs and 0.102 for SSRs). Cluster analysis of the ISSR and SSR results by the UPGMA method revealed two distinct groups correlated with the geographic origin of sampling. These results offer a new evidence for the theory of the origin of Hordeum vulgare L. PMID- 19304271 TI - The electrosensorial pore system of the cephalofoil in the four most common species of hammerhead shark (Elasmobranchii: Sphyrnidae) from the Southwestern Atlantic. AB - The laterally expanded head is the principal character distinguishing hammerhead sharks, and its morphology is important for interpreting their ontogeny and species diversity. Because their head shape changes during its ontogeny, it is vital to evaluate it in order to establish other taxonomical characteristics to correctly identify Sphyrna species. This study examines the distribution of electrosensorial pore regions on the ventral surface of the cephalofoil (VSC) in Sphyrna lewini, S. tiburo, S. tudes and S. zygaena from the Southwestern Atlantic Ocean. The pore distribution patterns in the VSC can distinguish these species. Use of those patterns, with the head shape, confirms the identification of the four most common species of hammerhead sharks in the Southwestern Atlantic. PMID- 19304272 TI - No need to open the jar: a comparative study of Magnetic Resonance Imaging results on fresh and alcohol preserved common carps (Cyprinus carpio (L. 1758), Cyprinidae, Teleostei). AB - Magnetic Resonance Imaging (MRI) examinations have been conducted both on fresh and alcohol-preserved common carps (Cyprinus carpio). The images have been compared to those of a sagittally-cut frozen animal of the same species. This work shows that the images obtained are globally similar, and that the MRI technique can be applied to investigate the gross anatomy of alcohol-preserved specimens without destroying them. Unfortunately, this kind of study does not provide precise enough anatomical data for small specimens (less than 10 cm in total length) with a 1 Tesla magnetic field. Nevertheless, leaving the specimen in the jar during MRI examinations does not affect the quality of the final images. PMID- 19304273 TI - Asbestos-induced lung diseases: an update. AB - Asbestos causes asbestosis (pulmonary fibrosis caused by asbestos inhalation) and malignancies (bronchogenic carcinoma and mesothelioma) by mechanisms that are not fully elucidated. Despite a dramatic reduction in asbestos use worldwide, asbestos-induced lung diseases remain a substantial health concern primarily because of the vast amounts of fibers that have been mined, processed, and used during the 20th century combined with the long latency period of up to 40 years between exposure and disease presentation. This review summarizes the important new epidemiologic and pathogenic information that has emerged over the past several years. Whereas the development of asbestosis is directly associated with the magnitude and duration of asbestos exposure, the development of a malignant clone of cells can occur in the setting of low-level asbestos exposure. Emphasis is placed on the recent epidemiologic investigations that explore the malignancy risk that occurs from nonoccupational, environmental asbestos exposure. Accumulating studies are shedding light on novel mechanistic pathways by which asbestos damages the lung. Attention is focused on the importance of alveolar epithelial cell (AEC) injury and repair, the role of iron-derived reactive oxygen species (ROS), and apoptosis by the p53- and mitochondria-regulated death pathways. Furthermore, recent evidence underscores crucial roles for specific cellular signaling pathways that regulate the production of cytokines and growth factors. An evolving role for epithelial-mesenchymal transition (EMT) is also reviewed. The translational significance of these studies is evident in providing the molecular basis for developing novel therapeutic strategies for asbestos related lung diseases and, importantly, other pulmonary diseases, such as interstitial pulmonary fibrosis and lung cancer. PMID- 19304274 TI - Bronchiolitis obliterans in lung transplantation: the good, the bad, and the future. AB - Lung transplantation remains the hope for many incurable pulmonary diseases, such as cystic fibrosis, pulmonary fibrosis, and chronic obstructive pulmonary disease. Remarkable progress has been made in improving outcomes, although the incidence of acute rejection remains more than 50% in the 1st year, and the 5 year graft survival is still less than 50% primarily because of the development of chronic rejection and graft dysfunction. Chronic rejection is characterized by the development of obliterative bronchiolitis in allografts and manifests as bronchiolitis obliterans syndrome in humans with no effective treatment. Previous studies support a role for alloreactive T cells in the development of bronchiolitis obliterans syndrome, but the specific mechanisms are unknown. One major stumbling block to research in the field of lung transplantation has been the lack of physiologic models to study the disease in the laboratory. We will review the current understanding of the immunology of the pathogenesis of obliterative bronchiolitis and will discuss exciting new advances from the laboratory as well as the implications for future research in lung transplantation. PMID- 19304276 TI - Cell and gene: from transduction to translation. PMID- 19304275 TI - Oxidized phospholipids in control of inflammation and endothelial barrier. AB - The levels of circulating oxidized phospholipids (OxPLs) become increased in chronic and acute pathologic conditions such as hyperlipidemia, atherosclerosis, increased intimamedia thickness in the patients with systemic Lupus erythematosus, vascular balloon injury, acute lung injury (ALI), and acute respiratory distress syndrome (ARDS). These pathologies are associated with inflammation and activation of endothelial cells. Depending on the biological context and the specific group of phospholipid oxidation products, OxPL may exhibit both proinflammatory and anti-inflammatory effects. This review will summarize the data showing a dual role of OxPL in modulation of chronic and acute inflammation as well as OxPL effects on pulmonary endothelial permeability. Recent reports show protective effects of OxPL in the models of endotoxin and ventilator-induced ALI and suggest a potential for using OxPL-derived cyclopenthenone-containing compounds with barrier-protective properties for drug design. These compounds may represent a new group of therapeutic agents for the treatment of lung syndromes associated with acute inflammation and lung vascular leak. PMID- 19304277 TI - Blood outgrowth endothelial cell migration and trapping in vivo: a window into gene therapy. AB - Human blood outgrowth endothelial cells (hBOECs) may be useful delivery cells for gene therapy. hBOECs have high expansion capacity and a stable phenotype. If incorporated into blood vessels, hBOECs could release therapeutic agents directly into the bloodstream. However, little is known about the lodging and homing of hBOECs in vivo. We examined the homing patterns of hBOECs in mice and explored extending cell-based factor VIII (FVIII) gene therapy from mice to larger animals. hBOECs were injected into NOD/SCID mice to determine where they localize, how localization changes over time, and if there were toxic effects on host organs. The presence of hBOECs in mouse organs was determined by quantitative polymerase chain reaction (qPCR) and immunofluorescence microscopy. hBOECs lodged most notably in mouse lungs at 3 h, but by 24 h, no differences were observed among 9 organs. The longevity of hBOECs was assessed up to 7 months in vivo. hBOECs expanded well and then reached a plateau in vivo. hBOECs from older cultures expanded equally well in vivo as younger hBOECs. hBOECs caused no noticeable organ toxicity up to 3 days after injection. When mice were pretreated with antibodies to E-selectin, P-selectin, or anti-alpha4 integrin prior to hBOEC injection, the number of hBOECs in lungs at 3 h was decreased. Preliminary studies that infused hemophilic dogs with autologous canine BOECs that overexpressed FVIII (B-domain deleted) showed improvement in whole blood clotting times (WBCTs). In conclusion, the survivability, expandability, and lack of toxicity of BOECs in vivo indicate that they may be valuable host cells for gene therapy. PMID- 19304278 TI - Atorvastatin effect on the distribution of high-density lipoprotein subfractions and human paraoxonase activity. AB - Human serum paraoxonase-1 (PON1) protects lipoproteins against oxidation by hydrolyzing lipid peroxides in oxidized low-density lipoprotein (LDL); therefore, it may protect against atherosclerosis. Changes in the ratio of high-density lipoprotein (HDL) subfractions may alter the stability and the antioxidant capacity of PON1. The aim of the study was to examine the effect of atorvastatin treatment on the distribution of HDL subfractions, LDL size, cholesteryl ester transfer protein (CETP), lecithin-cholesterol acyltransferase (LCAT), and PON1 activity. In all, 33 patients with type IIa and IIb hypercholesterolemia were involved in the study. LDL sizes and HDL subfractions were determined by gradient gel electrophoresis. CETP, LCAT, and PON1 activities were measured spectrophotometrically. Three months of treatment with atorvastatin 20 mg daily significantly increased the HDL3 (+8.13%) and decreased the HDL2a and HDL2b subfractions (-1.57% and -6.55%, respectively). The mean LDL size was significantly increased (+3.29%). The level of oxidized LDL was significantly decreased (-46.0%). The PON1 activity was augmented by the atorvastatin treatment (+5.0%). The CETP activity positively correlated with the HDL2b level and negatively correlated with the HDL3 and HDL2a levels. Atorvastatin alters the HDL subfractions, which may improve its antiatherogenic effect via enhancement of the PON1 activity. PMID- 19304279 TI - High-molecular-weight adiponectin does not predict cardiovascular events in patients with type 2 diabetes. AB - Low circulating high-molecular-weight (HMW) adiponectin might be associated with increased cardiovascular risk. This study aimed to investigate the relationship between HMW adiponectin and cardiovascular events in patients with type 2 diabetes mellitus (T2DM) with an adverse cardiovascular risk profile. The investigation took place in a specialized outpatient clinic for metabolic diseases and included 147 patients with T2DM following a cross-sectional and a prospective study protocol. Ninety patients had macrovascular disease at baseline defined as preexisting coronary artery disease, previous stroke, or peripheral artery disease. HMW adiponectin measured by enzyme-linked immunosorbent assay (Fujirebio, Tokyo, Japan) and routine clinical parameters were determined in all patients at baseline. The occurrence of new cardiovascular events (myocardial infarction, stroke, and all-cause mortality) during the follow-up period was evaluated. No significant correlations between traditional cardiovascular risk markers and HMW adiponectin could be detected. HMW adiponectin did not differ between subjects with and without macrovascular disease at baseline (3.5 [interquartile range [IQR]: 2.2-5.7] mg/L vs 4.0 [IQR: 2.5-7.1] mg/L). During a follow-up of 19.3 (IQR: 16-25) months, 61 endpoints (41 myocardial infarctions, 10 strokes, and 10 deaths) were observed. A 1-standard-deviation increment of log transformed HMW adiponectin was not significantly associated with the occurrence of cardiovascular events (Adjusted hazard ratio [HR]: 0.95; 95% confidence interval [CI]: 0.58-1.54; P = 0.835). In conclusion, HMW adiponectin was not related to present macrovascular disease and is not associated with future cardiovascular events in high-risk patients with T2DM. It is unlikely that HMW adiponectin has significant vasoprotective effects in these patients. PMID- 19304280 TI - Adrenal tumours and hormone excess. Editorial. PMID- 19304281 TI - [Malignant disease of the parotid gland]. PMID- 19304282 TI - [Laparoscopic liver surgery. A mature option?]. PMID- 19304283 TI - Induction of IL-6 and inhibition of IL-8 secretion in the human airway cell line Calu-3 by urban particulate matter collected with a modified method of PM sampling. AB - Exposure to particulate matter (PM) induces inflammatory cytokines. In the present study, we evaluated the secretion of IL-6 and IL-8 by an airway cell line exposed to PM with a mean aerodynamic size equal to or less than 10 or 2.5 microm (PM10 and PM2.5, respectively) collected in Mexico City, using a modified high volume sampling method avoiding the use of solvents or introducing membrane components into the samples. PM was collected on cellulose-nitrate (CN) membranes modified for collection on high-volume samplers. Composition of the particles was evaluated by particle-induced X-ray emission (PIXE) and scanning electron microscopy. The particles (10-160 microg/cm2) were tested on Calu-3 cells. Control cultures were exposed to LPS (10 ng/mL to 100 microg/mL) or silica (10 160 microg/cm2). IL-6 and IL-8 secretions were evaluated by ELISA. An average of 10 mg of PM was recovered form each cellulose-nitrate filter. No evidence of contamination from the filter was found. Cells exposed to PM10 presented an increase in the secretion of IL-6 (up to 400%), while IL-8 decreased (from 40% to levels below the detection limit). A similar but weaker effect was observed with PM2.5. In conclusion, our modified sampling method provides a large amount of urban PM free of membrane contamination. The urban particles induce a decrease in IL-8 secretion that contrasts with the LPS and silica effects. These results suggest that the regulation of IL-8 expression is different for urban particles (complex mixture containing combustion-related particles, soil and biologic components) than for biogenic compounds or pure mineral particles. PMID- 19304285 TI - Increased cell proliferation in experimentally induced endometriosis in rabbits. AB - OBJECTIVE: To characterize the pattern of cell proliferation and apoptosis of eutopic and ectopic endometrium in rabbits after endometrium implantation for the experimental induction of endometriosis. DESIGN: Animal experimental study. SETTING: Sector of experimental surgery. ANIMAL(S): Twenty-female New Zealand rabbits. INTERVENTION(S): All animals underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium, and fixation of tissue segment to the pelvic peritoneum. Two groups of 10 animals were sacrificed 4 and 8 weeks after endometriosis induction. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma determination. MAIN OUTCOME MEASURE(S): Cell proliferation and apoptosis were determined in the eutopic and ectopic endometrium, and the cell proliferation index (CPI) and apoptotic index (AI) were calculated as the number of labeled cells per 1,000 cells. The tissue homeostasis index was the CPI/AI ratio. Glands and stroma were analyzed separately. RESULT(S): The CPI for ectopic tissue was increased compared with eutopic tissue, but there was no difference in the ectopic lesions between 4 and 8 weeks of induction. Considering only the AI, eutopic and ectopic endometrium did not differ after 4 weeks, but differed significantly in glandular tissue after 8 weeks. The tissue homeostasis index revealed cell proliferation in these tissues, with a CPI/AI more than 1. CONCLUSION(S): Ectopic lesions seem to have a higher CPI than eutopic endometrium, with uncontrolled tissue growth occurring in induced endometriotic lesions. PMID- 19304286 TI - Uterine conservation despite severe sepsis in a case of placenta accreta first treated conservatively: 3-month delayed successful removal of the placenta. AB - OBJECTIVE: To report a case of successful delayed removal of a placenta accreta first treated conservatively. Secondary infectious complications can occur after conservative management in cases of placenta accreta, most often leading to hysterectomy. DESIGN: Case report. SETTING: A French teaching hospital. PATIENT(S): A 33-year-old woman. INTERVENTION(S): A healthy 33-year-old woman underwent Cesarean section for her first pregnancy. Diagnosis of placenta accreta was made at ultrasound scanning for her second pregnancy. She was first treated conservatively. Hysterectomy was planned 3 months after conservative treatment because of sepsis attributed to uterine retention. (Hysterotomy was first realized.)-?? MAIN OUTCOME MEASURE(S): Uterine conservation. RESULT(S): The placenta was easily and successfully removed with no subsequent bleeding. The uterus was sutured and conserved. CONCLUSION(S): In cases of delayed sepsis because of uterine retention after conservative treatment for placenta accreta, when medical treatment remains unsuccessful, manual removal of the placenta should be attempted. This approach might allow improved uterine conservation rates in women with placenta accreta treated conservatively. PMID- 19304287 TI - Management of Salmonella ovarian abscess in a patient with systemic lupus erythematosus. AB - OBJECTIVE: To report a patient with systemic lupus erythematosus (SLE) with persistent ovarian abscess infected with Salmonella. DESIGN: Case report. SETTING: Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital. PATIENT(S): A 35-year-old woman with a 10-year history of SLE had intermittent fever with chills that was refractory to empirical antibiotics. INTERVENTION(S): Ceftriaxone was administered on the basis of blood culture showing Salmonella group B infection and was followed by image-guided drainage. MAIN OUTCOME MEASURE(S): Removal of pelvic abscess. RESULT(S): Right ovarian abscess persisted despite percutaneous drainage. Laparoscopic removal of the encapsulated abscess was performed, and she remained well during her 6-month follow-up. CONCLUSION(S): Appropriate administration of antibiotics is the mainstay treatment for Salmonella infection complicated with SLE. Any localized abscess after prolonged antibiotics should be treated with surgical debridement to ensure complete eradication of infection. Conservative surgery with use of laparoscopy is an option in skilled personnel. PMID- 19304288 TI - Effects of metoclopramide-induced hyperprolactinemia on the prolactin receptor of murine endometrium. AB - OBJECTIVE: To evaluate the effects of metoclopramide-induced hyperprolactinemia on the prolactin receptor of murine endometrium. DESIGN: Experimental study using the RNA extraction to detect tissue prolactin receptor isoforms by reverse transcriptase polymerase chain reaction (RT-PCR). SETTING: University-based laboratory. ANIMAL(S): Seventy-two female swiss albino mice (Mus musculus), approximately 100 days old, were divided into six 12-animal groups: (GI) nonoophorectomized mice given vehicle; (GII) nonoophorectomized mice treated with metoclopramide; (GIII) oophorectomized mice treated with metoclopramide; (GIV) oophorectomized mice treated with metoclopramide and 17beta-estradiol; (GV) oophorectomized mice treated with metoclopramide and micronized progesterone; (GVI) oophorectomized mice treated with metoclopramide and a solution of 17beta estradiol and micronized progesterone. INTERVENTION(S): Drugs were administered for 50 days. Following euthanasia, the middle portions of the uterine horns were removed, sectioned, and immediately frozen for RT-PCR procedures. Blood was collected for the dosage of prolactin and serum estrogen and progesterone using radioimmune assay. MAIN OUTCOME MEASURE(S): Identification of uterine prolactin receptor isoforms. RESULT(S): The PRL receptor and its isoform L were identified only in GI (control group) and GII (metoclopramide), the two groups with nonoophorectomized animals. The amount of PRL receptor mRNA and that of its isoform L from GII were the largest. No other isoforms of the prolactin receptor were identified in any of the groups. CONCLUSION(S): Our results suggest that replacement of estrogen and progestin may not increase the mRNA of endometrial PRL receptor in metoclopromide-induced hyperprolactinemia in rats after castration. PMID- 19304289 TI - Controversies in the management of nonobstructive azoospermia. AB - The fertility potential of patients with nonobstructive azoospermia (NOA) depends on sperm extraction from the tissue sample and then in vitro fertilization with intracytoplasmic sperm injection (IVF/ICSI). Unfortunately, there is no consensus regarding predictors that can identify nonobstructive azoospermic men with a potentially high yield at the time of sperm extraction. This article analyzes two competing approaches to these patients: noninvasive and invasive. The noninvasive approach, based on clinical, laboratory, and ultrasonographic investigations, excludes from IVF/ICSI a significant number of patients owing to errors in predicting the presence of sufficient intratesticular spermatozoa. The invasive approach, with available percutaneous or surgical testicular biopsy techniques followed by morphologic examination and or sperm recovery, permits many patients with NOA to receive a favorable prognosis and therapeutic trial. However, the available testicular biopsy techniques are so variable that their performance parameters cannot be adequately compared. As a result, any progress in optimizing these techniques must involve delineation of specific selection criteria for each NOA patient. PMID- 19304290 TI - Feasibility of using gravimetry to measure excess adsorption effects of a binary solvent system in a reversed-phase high-performance liquid chromatography column. AB - A modified method for weighing HPLC columns filled with solvent is described. The method prevents the loss of traces of solvent from within the threads of the column. The method was tested by obtaining the weights of a C18 column filled with 10 different organic solvents, showing a standard deviation on the order of 0.1%. A plot of gross column weight versus solvent density showed excellent linearity. The method was then used to weigh a column filled with several acetonitrile-water mixtures. The gross column weights were lower than would have been predicted from the density of the acetonitrile-water mixtures. A likely explanation is the existence of an adsorbed acetonitrile-rich liquid on the surface of the C18 adsorbent, which caused the lower than expected weights due to the lower density of pure acetonitrile relative to the bulk mixtures. The volume of pure acetonitrile required for the observed weight discrepancy was calculated. Based on the surface area of the column adsorbent, values of micromoles acetonitrile per square meter of surface area were determined. The values showed reasonable agreement with values obtained from published adsorption isotherm studies. This suggests that pycnometry may be a useful technique for adsorption studies. The limitations of the technique are discussed. PMID- 19304291 TI - A new gas chromatography/mass spectrometry method for the simultaneous analysis of target and non-target organic contaminants in waters. AB - In this study we developed a GC-MS method for the analysis of priority pollutants, personal care products (PCPs) and other emerging contaminants in waters using large volume injection with backflushing. Analyses are performed in the SIM/scan mode, so that in addition to the targeted organic contaminants, this method allows the simultaneous screening of non-target compounds. The scan data are analysed using Deconvolution Reporting Software (DRS) which screens the results for 934 organic contaminants. Deconvolution helps identify contaminants that are buried in the chromatogram by co-extracted materials and significantly reduces chromatographic resolution requirements, allowing shorter analysis times. All compounds have locked retention times and we can continually update and extend the mass spectral library including new compounds. Linearity and limits of detection in SIM and full-scan mode were studied. Method detection limits (MDLs) in effluent wastewater ranged in most of the cases from 1 to 36 ng/L in SIM mode and from 4 to 66 ng/L in full-scan mode; while in river water from 0.4 to 14 and 2-29 ng/L in SIM and full-scan mode, respectively. We obtained a linearity of the calibration curves over two orders of magnitude. The method has been applied to the screening of a large number of organic contaminants--not only to a subset of targets--in urban wastewaters from different wastewater treatment plants and also in river waters. Most of the target compounds were detected at concentration levels ranging from 11 to 8697 ng/L and from 7 to 1861 ng/L in effluent wastewater and river waters, respectively. Additionally, a group of 12 new compounds were automatically identified using the AMDIS and NIST libraries. Other compounds, such as the 4-amino musk xylene, a synthetic fragrance metabolite, which was not included in the databases, but has been manually searched in the full-scan chromatograms. PMID- 19304292 TI - Characterization and evaluation of a macroporous adsorbent for possible use in the expanded bed adsorption of flavonoids from Ginkgo biloba L. AB - The suitability of the use of macroporous adsorbent Amberlite XAD7HP in expanded bed adsorption processes for the isolation of flavonoids from crude extracts of Ginkgo biloba L. has been assessed. The expansion and hydrodynamic properties of expanded beds were investigated and analyzed. The bed expansion as a function of operational fluid velocity was measured and correlated with the Richardson-Zaki equation. Theoretical predictions of the correlation parameters (the terminal settling velocity u(t) and exponent n) were improved by modifying equations in the literature. Residence time distributions (RTDs) were studied using acetone as a tracer. Three measures of liquid phase dispersion (the height equivalent of theoretical plate, Bodenstein number and axial distribution coefficient) were investigated and compared to values previously obtained with commercial EBA adsorbents developed for protein purification. A suitable bed expansion ratio was found to be 1.25 times the settled bed height, which occurred at a corresponding flow velocity of 183 cm/h. For an initial settled bed height of 42 cm, the mean residence time of liquid in the expanded bed was around 28 min. Under these flow conditions, the axial mixing coefficient D(ax) was 7.54 x 10(-6) m(2)/s and the Bodenstein number was 28; the number of theoretical plates (N) was 19 and the height equivalent of a theoretical plate (HETP) was 2.77 cm. Rutin trihydrate was used as a model flavonoid for the characterization of the adsorption properties of Amberlite XAD7HP. Adsorption was observed to reach equilibrium within 3 h with 70% of the adsorption capacity being achieved within 30 min. The estimated maximum equilibrium adsorption capacity for rutin was estimated to be 43.0 mg/(gresin) when the results were fitted to Langmuir isotherms. The adsorption performance was not seriously impaired by the physical presence of G. biloba leaf powders. Assessment of the kinetics of the adsorption of rutin revealed that the rate constant for adsorption was only reduced by 15% in the presence of leaf powders at a concentration of 50 mg/mL. The results demonstrated that Amberlite XAD7HP should be suitable for expanded bed adsorption of flavonoids from crude extracts of G. biloba L. PMID- 19304293 TI - The effects of self-generated synchronous and asynchronous visual speech feedback on overt stuttering frequency. AB - PURPOSE: Relatively recent research documents that visual choral speech, which represents an externally generated form of synchronous visual speech feedback, significantly enhanced fluency in those who stutter. As a consequence, it was hypothesized that self-generated synchronous and asynchronous visual speech feedback would likewise enhance fluency. Therefore, the purpose of this study was to investigate the effects of self-generated visual feedback (i.e., synchronous speech feedback with a mirror and asynchronous speech feedback via delayed visual feedback) on overt stuttering frequency in those who stutter. METHOD: Eight people who stutter (4 males, 4 females), ranging from 18 to 42 years of age participated in this study. Due to the nature of visual speech feedback, the speaking task required that participants recite memorized phrases in control and experimental speaking conditions so that visual attention could be focused on the speech feedback, rather than a written passage. During experimental conditions, participants recited memorized phrases while simultaneously focusing on the movement of their lips, mouth, and jaw within their own synchronous (i.e., mirror) and asynchronous (i.e., delayed video signal) visual speech feedback. RESULTS: Results indicated that the self-generated visual feedback speaking conditions significantly decreased stuttering frequency (Greenhouse-Geisser p=.000); post hoc orthogonal comparisons revealed no significant differences in stuttering frequency reduction between the synchronous and asynchronous visual feedback speaking conditions (p=.2554). CONCLUSIONS: These data suggest that synchronous and asynchronous self-generated visual speech feedback is associated with significant reductions in overt stuttering frequency. Study results were discussed relative to existing theoretical models of fluency-enhancement via speech feedback, such as the engagement of mirror neuron networks, the EXPLAN model, and the Dual Premotor System Hypothesis. Further research in the area of self-generated visual speech feedback, as well as theoretical constructs accounting for how exposure to a multi-sensory speech feedback enhances fluency, is warranted. LEARNING OUTCOMES: : Readers will be able to (1) discuss the multi sensory nature of fluency-enhancing speech feedback, (2) compare and contrast synchronous and asynchronous self-generated and externally generated visual speech feedback, and (3) compare and contrast self-generated and externally generated visual speech feedback. PMID- 19304294 TI - Paired associate learning in Chinese children with dyslexia. AB - A total of 82 Chinese 11- and 12-year-olds with and without dyslexia were tested on four paired associate learning (PAL) tasks, phonological awareness, morphological awareness, rapid naming, and verbal short-term memory in three different experiments. Experiment 1 demonstrated that children with dyslexia were significantly poorer in visual-verbal PAL than nondyslexic children but that these groups did not differ in visual-visual PAL performance. In Experiment 2, children with dyslexia had more difficulties in transferring rules to new stimuli in a rule-based visual-verbal PAL task as compared with children without dyslexia. Long-term retention of PAL was not impaired in dyslexic children across either experiment. In Experiment 3, rates of visual-verbal PAL deficits among children with dyslexia were all at or above 39%, the highest among all cognitive deficits tested. Moreover, rule-based visual-verbal PAL, in addition to morphological awareness and rapid naming ability, uniquely distinguished children with and without dyslexia even with other metalinguistic skills statistically controlled. Results underscore the importance of visual-verbal PAL for understanding reading impairment in Chinese children. PMID- 19304295 TI - Maternal inexperience as a risk factor of innate fear and PTSD-like symptoms in mice. AB - In laboratory rats and mice, differences in maternal care during the first week of life have been shown to exert long-lasting consequences on cognitive functioning and stress processing of the offspring. Such epigenetic programming is also assumed to play an important role in the transgenerational transmission of PTSD in humans. Here we studied whether even subtle within-subject differences in maternal care - caused by increasing mothering experience from the first to the second litter - can determine subsequent vulnerability for PTSD-like behaviour. To assess the influence of maternal experience on different components of fear, we analysed the adult male offspring of two subsequent litters (offspring 1, 2) from the same parental C57BL/6NCrl (B6N) and C57BL/6JOla (B6JOla) mice for (i) their innate anxiety behaviour on a modified hole board and (ii) their vulnerability to develop long-lasting PTSD-like fear symptoms ("hyperarousal", contextually conditioned fear) following perception of an inescapable foot shock. Increasing maternal experience reduced the animals' innate fear on the modified hole board (more exploration, less inhibition), the acute stress reaction to the shock and - one month after trauma - the levels of hyperarousal-like behaviour in the PTSD-prone B6N strain. In contrast, both acquisition and extinction of contextually conditioned fear were increased in the second offspring, representing cognitive flexibility. A factor analysis showed that innate fear, "hyperarousal" and conditioned fear represent independent behavioural dimensions. In conclusion, the present study identifies maternal inexperience as a risk factor for the development of PTSD-like symptoms. This effect - occurring in inbred mice on an almost identical genetic background - emphasizes the impact of epigenetic factors in PTSD-like behaviour. PMID- 19304296 TI - How much does hypertension affect cognition?: explained variance in cross sectional analysis of non-demented community-dwelling individuals in the SEARCH study. AB - Vascular pathology impairs cognition and impaired cognition increases the risk of dementia. Hypertension is arguably the vascular risk factor that can be reverted best. Here we estimated the effect magnitude of hypertension by determining the variance in cognition explained by systolic blood pressure (sBP) in non-demented community-dwelling individuals. We recruited 525 individuals (mean age 65, range 40-85) selected from the city registry of Muenster, Germany, measured cognitive performance with a comprehensive test battery and assessed vascular risk based on glycosylated hemoglobin, serum cholesterol, high sensitive C-reactive protein, body mass index, smoking pack years, and blood pressure. Including gender and education as well as the vascular risk factors, multiple linear regression analysis for different age groups showed that in midlife age groups systolic blood pressure explained up to 11% of the variance in cognitive performance. These findings suggest that in non-demented community-dwelling individuals hypertension may account for one tenth of cognitive impairment and thus for an increased risk for dementia. PMID- 19304297 TI - Anthrax meningoencephalitis--declining trends in an uncommon but catastrophic CNS infection in rural Tamil Nadu, South India. AB - INTRODUCTION: Anthrax, a cattle-born zoonosis has been a serious infectious disease and its meningoencephalitic form remains a rapidly fatal illness even now. AIM: The aim of this paper is to evaluate the incidence and clinical profile of anthrax meningoencephalitis admitted to a teaching hospital predominantly serving a rural population in South India. MATERIALS AND METHODS: We made a systematic study of the case records of patients with microbiologically confirmed diagnosis of anthrax meningoencephalitis admitted over a 20-year period. We searched the internet and office records for the anthrax outbreaks and the preventive strategies in place in India. RESULTS: The admissions occurred in two clusters, four during 1992-1994 and six in 1998-2000; with no further detection unto August 2008. All patients were adult males with agriculture related occupation. Three of the 10 patients had no evidence of primary focus of infection. Majority were in coma at admission and had documented evidence of septicemia. CSF was haemorrhagic and death was the uniform outcome despite high dose intravenous penicillin G; maximum duration of hospital survival being 48 h. COMMENT: Anthrax is a rare, but catastrophic cause of meningoencephalitis. Improvement in education and life styles as well as livestock vaccination in rural areas appear to have effectively decreased the incidence of this dreaded acute zoonosis in the South Indian states of Tamilnadu and Puducherry. PMID- 19304298 TI - Prediction of long-term outcome by percent improvement after the first day of thrombolytic treatment in stroke patients. AB - BACKGROUND: We investigated a method for assessing early improvement and predictive factors of early and late outcomes in patients receiving thrombolytic therapy. METHODS: A total of 160 consecutive patients who received thrombolytic therapy were included in the study. Using National Institutes of Health Stroke Scale (NIHSS) scores, percent improvement [(baseline NIHSS score-24-hour NIHSS score)/baseline NIHSS score x 100] was calculated and compared with delta (baseline NIHSS score-24-hour NIHSS score) and with major neurological improvement (MNI, NIHSS score of 0-1 or >or=8 point improvement at 24 h) by receiver operating characteristic (ROC) curve analysis. Finally, we investigated the independent predictors of improvement at 24 h after the thrombolytic therapy and of favorable 3-month outcome (modified Rankin scale score 0-2). RESULTS: By pairwise comparison of ROC curves, percent improvement was stronger than delta (p=0.004) and MNI (p<0.001) in predicting long-term outcome. First day improvement (FDI), defined as greater than 20% improvement, was a strong predictor of favorable 3-month outcome (OR 12.55, 95% CI 5.41-29.10). Recanalization (OR 3.30, 95% CI 1.28-8.45), absence of carotid T occlusion (OR 0.09, 95% CI 0.02-0.42) and hemorrhagic transformation (OR 0.25, 95% CI 0.09 0.73) were independent predictors of FDI. Independent predictors of favorable 3 month outcome were FDI, current smoking, absence of carotid T occlusion and hemorrhagic transformation. CONCLUSIONS: Percent improvement at 24 h after thrombolytic therapy is a useful surrogate marker for predicting the long-term outcome. Our findings highlight the importance of early stroke management. PMID- 19304299 TI - Neuroprotective effect of treatment with galantamine and choline alphoscerate on brain microanatomy in spontaneously hypertensive rats. AB - The present study was designed to assess if treatment with acetylcholinesterase inhibitor galantamine and the cholinergic precursor choline alphoscerate (alpha glyceryl-phosphoryl-choline) alone or in association has any protective effect on brain microanatomy in spontaneously hypertensive rats (SHR) used as an animal model of vascular dementia (VaD). Thirty-two-week-old SHR and age-matched normotensive Wistar Kyoto (WKY) rats were left untreated or treated for 4 weeks with an oral dose of 3 mg/kg/day of galantamine, of 100 mg/kg/day of choline alphoscerate or their association. The number of neurons and of glial fibrillary acidic protein (GFAP) immunoreactive astrocytes, phosphorylated neurofilament, and microtubule associated protein-2 (MAP-2) and aquaporin-4 (AQP-4) was assessed by quantitative microanatomical and immunohistochemical techniques. In SHR, the number of neurons of frontal cortex, of the CA1 subfield of hippocampus and of dentate gyrus was decreased compared to WKY rats. Astrogliosis, breakdown of phosphorylated neurofilament, unchanged MAP-2 and altered AQP-4 expression were found as well. Both galantamine and choline alphoscerate countered nerve cell loss. Choline alphoscerate but not galantamine decreased astrogliosis and restored expression of AQ-4. Galantamine countered to a greater extent than choline alphoscerate phosphorylated neurofilament breakdown. The two drugs in association displayed a more remarkable effect. This study confirms a neuroprotective effect of galantamine in SHR and indicates a neuroprotective role of choline alphoscerate in the same model. A wider neuroprotective effect of the cholinergic inhibitor/precursor association was observed. These findings suggest to assess the activity of this cholinergic association in clinical trials. PMID- 19304300 TI - [Functional anatomy of the cochlear nerve and the central auditory system]. AB - The auditory pathways are a system of afferent fibers (through the cochlear nerve) and efferent fibers (through the vestibular nerve), which are not limited to a simple information transmitting system but create a veritable integration of the sound stimulus at the different levels, by analyzing its three fundamental elements: frequency (pitch), intensity, and spatial localization of the sound source. From the cochlea to the primary auditory cortex, the auditory fibers are organized anatomically in relation to the characteristic frequency of the sound signal that they transmit (tonotopy). Coding the intensity of the sound signal is based on temporal recruitment (the number of action potentials) and spatial recruitment (the number of inner hair cells recruited near the cell of the frequency that is characteristic of the stimulus). Because of binaural hearing, commissural pathways at each level of the auditory system and integration of the phase shift and the difference in intensity between signals coming from both ears, spatial localization of the sound source is possible. Finally, through the efferent fibers in the vestibular nerve, higher centers exercise control over the activity of the cochlea and adjust the peripheral hearing organ to external sound conditions, thus protecting the auditory system or increasing sensitivity by the attention given to the signal. PMID- 19304301 TI - [Functional anatomy of the glossopharyngeal, vagus, accessory and hypoglossal cranial nerves]. AB - The glossopharyngeal nerve and the vagus nerve are mixed nerves containing sensory, gustatory, motor and autonomous fibers (parasympathetic). The glossopharyngeal nerve has mainly visceral afferent fibers from the tongue and pharynx, gustatory fibers from the posterior third of the tongue, parasympathetic afferent fibers from carotid sinus and carotid glomus, parasympathetic efferent fibers for the parotid gland and motor fibers for the muscles of the pharynx. The vagus nerve contains mostly visceral afferent fibers from laryngeal, intrathoracic and abdominal organs, parasympathetic efferent fibers for these intrathoracic and abdominal organs and motor fibers to the pharyngeal and laryngeal muscles. The accessory nerve is divided into two branches, different in their origins and their functions: the cranial portion joins the motor fibers of the vagus nerve, to form the recurrent laryngeal nerve, whereas the spinal portion innervates the sternocleidomastoid muscle and the trapezius muscle. Finally, the hypoglossal nerve is the main motor nerve of the tongue. PMID- 19304302 TI - Driving force of acoustic streaming caused by aperiodic sound beam in unbounded volumes. AB - Instantaneous driving force of acoustic streaming in the thermoviscous medium is the subject of investigation. Dynamic equation of the Eulerian streaming velocity is a result of splitting the hydrodynamic equations into acoustic and non acoustic parts. The acoustic force represents a sum of three parts, one is the classic one, which being averaged over the sound period coincides with the well known expression. The second one is connected to the periodicity of the sound, it becomes exact zero after averaging for the strictly periodic sound but is not zero for other acoustic wave. The last term originates from the sound divergence. All terms are nonlinear and proportional to the overall attenuation. The consistent comparative analysis of both formula for quasi-periodic and modulated sound is proceeded. PMID- 19304303 TI - Deep sequencing analysis of RNAs from a grapevine showing Syrah decline symptoms reveals a multiple virus infection that includes a novel virus. AB - In a search for viruses associated with decline symptoms of Syrah grapevines, we have undertaken an analysis of total plant RNA sequences using Life Sciences 454 high-throughput sequencing. 67.5 megabases of sequence data were derived from reverse-transcribed cDNA fragments, and screened for sequences of viral or viroid origin. The data revealed that a vine showing decline symptoms supported a mixed infection that included seven different RNA genomes. Fragments identified as derived from viruses or viroids spanned a approximately ten thousand fold range in relative prevalence, from 48,278 fragments derived from Rupestris stem pitting associated virus to 4 fragments from Australian grapevine viroid. 1527 fragments were identified as derived from an unknown marafivirus. Its complete genome was sequenced and characterized, and an RT-PCR test was developed to analyze its field distribution and to demonstrate its presence in leafhoppers (vector for marafiviruses) collected from diseased vines. Initial surveys detected a limited presence of the virus in grape-growing regions of California. PMID- 19304304 TI - Restriction of HIV-1 by APOBEC3G is cytidine deaminase-dependent. AB - Cytidine deamination is the primary mechanism by which APOBEC3G restricts HIV-1; however, several studies have reported that APOBEC3G also inhibits virus replication via a mechanism that is independent of deamination. Using active site APOBEC3G mutants, we have re-evaluated the biological relevance of deaminase independent APOBEC3G-mediated restriction of HIV-1. APOBEC3G proteins with Glu- >Ala mutations in AS1, AS2 or AS1 and AS2 were stably expressed at physiological levels in CEM-SS T cells and 293T cells and the ability of the cells to support Deltavif HIV-1 replication was then tested. The AS2 and AS1/AS2 mutants were packaged efficiently into virions but in single-cycle or multi-cycle HIV-1 replication assays, were found to lack antiviral activity. The AS1 mutant, which retained deaminase activity, maintained near wild-type antiviral function. To determine the potency of APOBEC3G antiviral activity, cell lines were established that that expressed low levels of wild-type APOBEC3G and generated virions that contained as few as 1-2 APOBEC3G molecules. Even at very low copy number, APOBEC3G caused a significant reduction in infectivity, suggesting that a single molecule of packaged APOBEC3G inactivates the virus. The high potency of APOBEC3G is consistent with a catalytic mechanism of restriction in which a single molecule can induce a string of mutations but difficult to reconcile with a deaminase-independent, non-catalytic mechanism. Analysis of the reverse transcript sequences showed that the G-->A mutations were clustered, likely reflecting the action of single APOBEC3G molecules acting processively. We conclude that cytidine deamination is the mechanism by which APOBEC3G restricts HIV-1. PMID- 19304305 TI - Induction of T helper 3 regulatory cells by dendritic cells infected with porcine reproductive and respiratory syndrome virus. AB - Delayed development of virus-specific immune response has been observed in pigs infected with the porcine reproductive and respiratory syndrome virus (PRRSV). Several studies support the hypothesis that the PRRSV is capable of modulating porcine immune system, but the mechanisms involved are yet to be defined. In this study, we evaluated the induction of T regulatory cells by PRRSV-infected dendritic cells (DCs). Our results showed that PRRSV-infected DCs significantly increased Foxp3(+)CD25(+) T cells, an effect that was reversible by IFN-alpha treatment, and this outcome was reproducible using two distinct PRRSV strains. Analysis of the expressed cytokines suggested that the induction of Foxp3(+)CD25(+) T cells is dependent on TGF-beta but not IL-10. In addition, a significant up-regulation of Foxp3 mRNA, but not TBX21 or GATA3, was detected. Importantly, our results showed that the induced Foxp3(+)CD25(+) T cells were able to suppress the proliferation of PHA-stimulated PBMCs. The T cells induced by the PRRSV-infected DCs fit the Foxp3(+)CD25(+) T helper 3 (Th3) regulatory cell phenotype described in the literature. The induction of this cell phenotype depended, at least in part, on PRRSV viability because IFN-alpha treatment or virus inactivation reversed these effects. In conclusion, this data supports the hypothesis that the PRRSV succeeds to establish and replicate in porcine cells early post-infection, in part, by inducing Th3 regulatory cells as a mechanism of modulating the porcine immune system. PMID- 19304306 TI - The 8ab protein of SARS-CoV is a luminal ER membrane-associated protein and induces the activation of ATF6. AB - The 8ab protein of SARS-CoV is a group-specific accessory protein, which is lost when the virus was transmitted from animals to humans due to a 29-nucleotide deletion in the ORF8ab region. Here we found that 8ab protein is associated with ER membrane at luminal surface. 8ab protein was found to up-regulate the synthesis of endogenous ER-resident chaperons involved in protein folding through the activation of the transcription factor ATF6, while it showed no effect on the CHOP induction and XBP1 splicing associated with the unfolded protein response (UPR). When ectopically expressed in mammalian cells, 8ab induced the proteolysis of ATF6 and the translocation of its cleaved DNA-binding and transcription activation domains from the ER to the nucleus. Finally, we showed that 8ab binds to the luminal domain of ATF6. These findings suggest that 8ab could modulate the UPR by activating ATF6 to facilitate protein folding and processing. Thus, the loss of 8ab in SARS-CoV through viral evolution in animals may play a role in its pathogenicity. PMID- 19304307 TI - Single-particle cryo-electron microscopy of Rift Valley fever virus. AB - Rift Valley fever virus (RVFV; Bunyaviridae; Phlebovirus) is an emerging human and veterinary pathogen causing acute hepatitis in ruminants and has the potential to cause hemorrhagic fever in humans. We report a three-dimensional reconstruction of RVFV vaccine strain MP-12 (RVFV MP-12) by cryo-electron microcopy using icosahedral symmetry of individual virions. Although the genomic core of RVFV MP-12 is apparently poorly ordered, the glycoproteins on the virus surface are highly symmetric and arranged on a T=12 icosahedral lattice. Our RVFV MP-12 structure allowed clear identification of inter-capsomer contacts and definition of possible glycoprotein arrangements within capsomers. This structure provides a detailed model for phleboviruses, opens new avenues for high resolution structural studies of the bunyavirus family, and aids the design of antiviral diagnostics and effective subunit vaccines. PMID- 19304308 TI - Identification of a lipid kinase as a host factor involved in hepatitis C virus RNA replication. AB - A functional screen of an adenovirus-delivered shRNA library that targets approximately 4500 host genes was performed to identify cellular factors that regulate hepatitis C virus (HCV) sub-genomic RNA replication. Seventy-three hits were further examined by siRNA oligonucleotide-directed knockdown, and silencing of the PI4KA gene was demonstrated to have a significant effect on the replication of a HCV genotype 1b replicon. Using transient siRNA oligonucleotide transfections and stable shRNA knockdown clones in HuH-7 cells, the PI4KA gene was shown to be essential for the replication of all HCV genotypes tested (1a, 1b and 2a) but not required for bovine viral diarrhea virus (BVDV) RNA replication. PMID- 19304309 TI - Chlorination disinfection by-products, public health risk tradeoffs and me. AB - Since 1974 when trihalomethanes (THMs) were first reported as disinfection by products (DBPs) in drinking water, there has been an enormous research effort directed at understanding how DBPs are formed in the chlorination or chloramination of drinking water, how these chlorination DBPs can be minimized and whether they pose a public health risk, mainly in the form of cancer or adverse reproductive outcomes. Driven by continuing analytical advances, the original DBPs, the THMs, have been expanded to include over 600 DBPs that have now been reported in drinking water. The historical risk assessment context which presumed cancer could be mainly attributed to exposure to environmental carcinogens played a major role in defining regulatory responses to chlorination DBPs which, in turn, strongly influenced the DBP research agenda. There are now more than 30 years of drinking water quality, treatment and health effects research, including more than 60 epidemiology studies on human populations, directed at the chlorination DBP issue. These provide considerable scope to reflect on what we know now, how our understanding has changed, what those changes mean for public health risk management overall and where we should look to better understand and manage this issue in the future. PMID- 19304310 TI - A simple dermal absorption model: derivation and application. AB - This paper deals with the derivation of a QSAR for the estimation of: the skin permeation coefficient from aqueous solutions in cm h(-1), the stratum corneum/water partition coefficient. These QSARs enable the estimation of: the aqueous permeation coefficient in cm h(-1), the maximum dermal absorption in mg cm(-2) h(-1) from a saturated aqueous solution at steady state, the lag time in hours (h), the diffusivity of a substance in the stratum corneum in cm2 h(-1). By using the independent variables: the log(octanol/water partition coefficient), the molecular weight,the water solubility. The estimated maximum dermal absorption and the lag time were compared with some recent measured data of substances, which were not used for developing the QSARs. The estimates were generally in the same order of magnitude as the measured absorption and lag time. These QSARs are recommended for risk assessment of chemicals in the scope of the European REACH legislation. PMID- 19304311 TI - Emissions of PCDD/Fs from municipal solid waste incinerators in China. AB - Gas emission of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) from 19 commercial municipal solid waste (MSW) incinerators in China are investigated. The emission concentrations of PCDD/Fs were 0.042-2.461 ng TEQ Nm(-3) with an average value of 0.423 ng TEQ Nm(-3). The emissions of PCDD/Fs from 16 MSW incinerators were below the MEP regulation level (1.0 ng I-TEQ Nm(-3)), while only six systems have the dioxin emission levels below the limit established by the European Union Directive of 0.1 ng I-TEQ Nm( 3). The emission factors of PCDD/Fs from 19 MSW incinerators were calculated to be 0.169-10.72 microg TEQ ton(-1) MSW with an average value of 1.728 microg I-TEQ ton(-1) MSW. The total amount of PCDD/Fs emitted from MSW incinerators to the atmosphere in China was estimated to 19.64 g TEQ year(-1) in 2006. PMID- 19304312 TI - Semi-empirical estimation of organic compound fugacity ratios at environmentally relevant system temperatures. AB - Fugacity ratios of organic compounds are used to calculate (subcooled) liquid properties, such as solubility or vapour pressure, from solid properties and vice versa. They can be calculated from the entropy of fusion, the melting temperature, and heat capacity data for the solid and the liquid. For many organic compounds, values for the fusion entropy are lacking. Heat capacity data are even scarcer. In the present study, semi-empirical compound class specific equations were derived to estimate fugacity ratios from molecular weight and melting temperature for polycyclic aromatic hydrocarbons and polychlorinated benzenes, biphenyls, dibenzo[p]dioxins and dibenzofurans. These equations estimate fugacity ratios with an average standard error of about 0.05 log units. In addition, for compounds with known fusion entropy values, a general semi empirical correction equation based on molecular weight and melting temperature was derived for estimation of the contribution of heat capacity differences to the fugacity ratio. This equation estimates the heat capacity contribution correction factor with an average standard error of 0.02 log units for polycyclic aromatic hydrocarbons, polychlorinated benzenes, biphenyls, dibenzo[p]dioxins and dibenzofurans. PMID- 19304313 TI - A distance-decay variable selection strategy for land use regression modeling of ambient air pollution exposures. AB - Land use regression (LUR) has emerged as an effective and economical means of estimating air pollution exposures for epidemiological studies. To date, no systematic method has been developed for optimizing the variable selection process. Traditionally, a limited number of buffer distances assumed having the highest correlations with measured pollutant concentrations are used in the manual stepwise selection process or a model transferred from another urban area. In this paper we propose a novel and systematic way of modeling long-term average air pollutant concentrations through "A Distance Decay REgression Selection Strategy" (ADDRESS). The selection process includes multiple steps and, at each step, a full spectrum of correlation coefficients and buffer distance decay curves are used to select a spatial covariate of the highest correlation (compared to other variables) at its optimized buffer distance. At the first step, the series of distance decay curves is constructed using the measured concentrations against the chosen spatial covariates. A variable with the highest correlation to pollutant levels at its optimized buffer distance is chosen as the first predictor of the LUR model from all the distance decay curves. Starting from the second step, the prediction residuals are used to construct new series of distance decay curves and the variable of the highest correlation at its optimized buffer distance is chosen to be added to the model. This process continues until a variable being added does not contribute significantly (p>0.10) to the model performance. The distance decay curve yields a visualization of change and trend of correlation between the spatial covariates and air pollution concentrations or their prediction residuals, providing a transparent and efficient means of selecting optimized buffer distances. Empirical comparisons suggested that the ADDRESS method produced better results than a manual stepwise selection process of limited buffer distances. The method also enables researchers to understand the likely scale of variables that influence pollution levels, which has potentially important ramifications for planning and epidemiological studies. PMID- 19304314 TI - Swirling flow created in a glass tube suppressed platelet adhesion to the surface of the tube: its implication in the design of small-caliber arterial grafts. AB - To test the hypothesis that intentionally inducing swirling blood flow in a small caliber arterial graft can suppress acute thrombus formation by affecting the adhesion of platelets to the internal surface of the graft, an experimental comparative study was designed to investigate the effect of swirling flow on the adhesion and activation of platelets in a straight glass tube coated with calf skin type I collagen. The experimental results showed that when compared with the normal flow, the swirling flow generated in the test tube significantly reduced the platelet adhesion to the surface of the test tube. Different from normal flow condition under which platelet adhesion increased simply with decreasing wall shear stress, the platelet adhesion density under swirling flow condition remained almost unchanged along the first 5 mm section of the tube in which the flow had relatively high rotation strength, even though the wall shear stress in this section of the tube dropped drastically. This suggests that when the swirling flow in the tube was strong enough, platelet adhesion was dominantly affected by the swirling flow itself, wall shear stress was secondary. The results also showed that there was no significant difference in the activation of platelets between the spiral flow group and the normal flow group. The present study therefore suggests that intentionally introducing swirling flow in small caliber arterial grafts has no adverse effect on platelet activation and may indeed be a solution to improving the patency of the grafts by suppressing acute thrombus formation. PMID- 19304315 TI - Optimization of the concentration of photo-initiator in a one-step self-etch adhesive. AB - OBJECTIVES: The objective of this study was to determine the optimal concentration of photo-initiator (camphorquinone) in an experimental one-step self-etch adhesive and to investigate the role of the photo-initiator. METHODS: Seven experimental one-step adhesives with a varying amount of camphorquinone ranging from 0 to 5.25wt% were prepared. Their micro-tensile bond strength to enamel and dentin was determined. In addition, the bond strength was also determined when the adhesive was not light-cured prior to the application of the composite. SEM and TEM were used for further evaluation of the resultant interfacial ultrastructure. RESULTS: The bond strength to enamel was not influenced by the amount of photo-initiator, whereas the bond strength to dentin dropped significantly when concentrations below 0.35wt% camphorquinone were used. Besides phase-separation droplets, electron microscopy revealed the presence of many small droplets at the bottom of the adhesive layer when the adhesive contained no or only a low concentration of initiator, or when the adhesive was not light-cured. SIGNIFICANCE: Since polymerization is severely hampered by oxygen inhibition in thin layers, one-step self-etch adhesives depend greatly on the polymerization of the first layer of lining composite to achieve their ultimate mechanical strength. Consequently, the bond strength to enamel is not influenced by the amount of photo-initiator, but on dentin, bond strength is compromised by droplets, probably due to water absorption, and additionally by the negative effect of water on polymerization and by continuing demineralization of unpolymerized acidic monomers. Overall, it was found that minimally 0.7wt% camphorquinone was needed. PMID- 19304316 TI - Developmental changes and gender differences in adolescents' perceptions of friendships. AB - This five-wave study aims to investigate the development of adolescents' perceptions of support, negative interaction, and power in best friendships from ages 12 to 20 years. Furthermore, gender differences and linkages between the three dimensions are explored. A total of 593 early adolescents (53.6% boys) and 337 middle adolescents (43.3% boys) participated. A multigroup multivariate accelerated growth curve showed an increase of support for both boys and girls. Negative interaction was found to temporarily increase and then decrease for boys, while remaining stable for girls. Power temporarily increased for boys and decreased for girls. Results indicated that (1) friendships become more supportive during adolescence, (2) power issues are more prominent in friendships of boys and more powerful peers are perceived as more supportive by boys but not by girls, and (3) friendships of boys show a lagged development towards more equality. PMID- 19304317 TI - Hydroxyapatite nano and microparticles: correlation of particle properties with cytotoxicity and biostability. AB - Synthetic colloid and gel hydroxyapatite (HA) nanoparticles (NPs) were spray dried to form microparticles (MPs). These are intended for use as slow release vaccine vectors. The physico-chemical properties of gel and colloid NPs and MPs were compared to those of HA obtained commercially. Their cytotoxicity to human monocytes'-derived macrophages (HMMs) was assessed in vitro using a range of techniques. These included the MTT assay, LDH leakage and a confocal based live dead cell assay. Cytotoxicity differed significantly between preparations, with the suspended gel preparation being the most toxic (31-500 microg/ml). Other preparations were also toxic but only at higher concentrations (>250 microg/ml). Transmission electron microscopy (TEM) and stereology showed variable cellular uptake and subsequent dissolution of the various forms of HA. We have demonstrated that HA particle toxicity varied considerably and that it was related to their physico-chemical properties. Cell death correlated strongly with particle load. The intracellular dissolution of particles as a function of time in HMM suggests that increased cytoplasmic calcium load is likely to be the cause of cell death. Some HA NPs eluded the phagocytic pathway and a few were even seen to enter the nuclei through nuclear pores. PMID- 19304318 TI - Calcium phosphate nanoparticles as efficient carriers for photodynamic therapy against cells and bacteria. AB - Calcium phosphate nanoparticles were surface-functionalized with different polymers, and photosensitizers were incorporated into this layer. The charge was adjusted by choosing the appropriate polymer. Methylene blue and 5,10,15,20 tetrakis(3-hydroxyphenyl)porphyrin (mTHPP) were used as photosensitizers. The particles showed a good performance with HIG-82 synoviocytes. For J774A.1 macrophages, they were toxic also in the dark, probably due to a lethal uptake of calcium. For HT29 epithelial cells, a moderate activity was observed. A good photoxicity was observed against the bacterial strain Staphylococcus aureus (Gram positive), both with positively and negatively charged nanoparticles loaded with mTHPP. Against Pseudomonas aeruginosa (Gram-negative), good photoxicity was observed only with positively charged nanoparticles loaded with mTHPP. At higher concentrations, methylene blue-loaded nanoparticles were active against S. aureus. Thus, it is possible to prepare a water-dispersable system of dye-loaded calcium phosphate nanoparticles, but the efficiency depends on a number of parameters, e.g. particle charge, kind of polymer, and cell culture medium (e.g. the presence of proteins). PMID- 19304319 TI - The augmentation of intracellular delivery of peptide therapeutics by artificial protein transduction domains. AB - Protein transduction domains (PTDs), such as HIV-derived Tat, have been successfully used as functional biomaterials for intracellular delivery of anti cancer macromolecular drugs (protein, peptides, and oligonucleotides). Although there were therefore great expectations regarding the therapeutic potential of PTDs for the development of anti-cancer therapeutics, their clinical application so far has been extremely limited because of the relatively high concentrations required to mediate any effects on cancer cells in vitro or in vivo. In this context, improving the transduction efficiency of PTDs using phage display-based molecular evolution techniques may be useful for creating artificial PTDs with high efficiency and safety. Here, we report an evaluation of transduction efficiency and toxicity of such artificial PTDs (designated mT02 and mT03) compared with Tat. The internalization of mT02 was the most rapid and efficient by a mechanism different from the usual macropinocytosis. Furthermore, we found that artificial PTDs fused with survivin antagonistic peptide potentiate tumor cell-cytostatic activity. Thus, the results of this work provide new insights for designing new-generation peptide therapeutics for a wide variety of cancers as well as those expressing survivin. PMID- 19304320 TI - Simultaneous delivery of doxorubicin and gemcitabine to tumors in vivo using prototypic polymeric drug carriers. AB - Copolymers of N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic and well characterized polymeric drug carriers that have been broadly implemented in the delivery of anticancer therapeutics. To demonstrate that polymers, as liposomes, can be used for simultaneously delivering multiple chemotherapeutic agents to tumors in vivo, we have synthesized and evaluated an HPMA-based polymer-drug conjugate carrying 6.4wt% of gemcitabine, 5.7wt% of doxorubicin and 1.0mol% of tyrosinamide (to allow for radiolabeling). The resulting construct, i.e. poly(HPMA-co-MA-GFLG-gemcitabine-co-MA-GFLG-doxorubicin-co-MA-TyrNH(2)), was termed P-Gem-Dox, and was shown to effectively kill cancer cells in vitro, to circulate for prolonged period of time, to localize to tumors relatively selectively, and to inhibit tumor growth. As compared to control regimens, P-Gem Dox increased the efficacy of the combination of gemcitabine and doxorubicin without increasing its toxicity, and it more strongly inhibited angiogenesis and induced apoptosis. These findings demonstrate that passively tumor-targeted polymeric drug carriers can be used for delivering two different chemotherapeutic agents to tumors simultaneously, and they thereby set the stage for more elaborate analyses on the potential of polymer-based multi-drug targeting. PMID- 19304321 TI - Microfluidic system for formation of PC-3 prostate cancer co-culture spheroids. AB - The niche microenvironment in which cancer cells reside plays a prominent role in the growth of cancer. It is therefore imperative to mimic the in vivo tumor niche in vitro to better understand cancer and enhance development of therapeutics. Here, we engineer a 3D metastatic prostate cancer model that includes the types of surrounding cells in the bone microenvironment that the metastatic prostate cancer cells reside in. Specifically, we used a two-layer microfluidic system to culture 3D multi-cell type spheroids of fluorescently labeled metastatic prostate cancer cells (PC-3 cell line), osteoblasts and endothelial cells. This method ensures uniform incorporation of all co-culture cell types into each spheroid and keeps the spheroids stationary for easy tracking of individual spheroids and the PC-3's residing inside them over the course of at least a week. This culture system greatly decreased the proliferation rate of PC-3 cells without reducing viability and may more faithfully recapitulate the in vivo growth behavior of malignant cancer cells within the bone metastatic prostate cancer microenvironment. PMID- 19304322 TI - The mechanical properties and bioactivity of poly(methyl methacrylate)/SiO(2)-CaO nanocomposite. AB - The mechanical properties and bioactivity of poly(methyl methacrylate)/SiO(2)-CaO nanocomposite were investigated using dimethyldiethoxysilane (DMDES) and tetraethoxysilane (TEOS), which could produce two and four siloxane linkages, respectively, after a sol-gel reaction. Methyl methacrylate was co-polymerized with 3-(trimethoxysilyl)propyl methacrylate and then co-condensed with DMDES (specimen D) and TEOS (specimen T), respectively, with calcium nitrate tetrahydrate under acidic conditions. The fracture toughness of specimen D was much improved compared to that of specimen T, whereas its fracture strength, hardness, and apatite-forming ability in simulated body fluid (SBF) were slightly decreased. The improved fracture toughness of specimen D without losing apatite forming ability was explained by the decrease of siloxane linkage numbers and the introduction of alkyl groups in silica structure because covalently bonded siloxane linkages produce hard and brittle fracture behavior in the nanocomposite while the alkyl groups help to make the silica as linear chain structure. The practical implication of these results is that this new nanocomposite can be applied to the filler materials for bone cement and dental composite resin because of its good bioactivity and improved mechanical properties. PMID- 19304323 TI - GATA-2 L359 V mutation is exclusively associated with CML progression but not other hematological malignancies and GATA-2 P250A is a novel single nucleotide polymorphism. AB - Chronic myeloid leukemia (CML) progression is characterized by occurrence of new cytogenetic and molecular abnormalities. In the previous study, we have shown the important role of GATA-2 L359 V mutation in CML progression. To further ascertain the truth of transcription factor GATA-2 in hematological malignancies, we expanded our study to GATA-2 full length by directly sequencing and applied MassARRAY assay into GATA-2 L359 V mutation analysis. Finally, no GATA-2 L359 V mutation was found in 270 acute myeloid leukemia, 30 myelodysplastic syndrome, 50 acute lymphoblastic leukemia, 12 chronic lymphocytic leukemia, 40 CML chronic phase and 286 BCR/ABL negative myeloproliferative disorders except CML blast crisis. A new variation of GATA-2 resulted in P250A change was identified, which was not found to have statistical difference between patients with hematological malignancies and healthy control. Hence, we concluded GATA-2 L359 V is exclusively associated with CML progression but not other hematological malignancies and P250A is a new single nucleotide polymorphism. PMID- 19304324 TI - Lenalidomide-induced durable hematological and cytogenetic remission in del(5q) associated de novo acute myeloid leukemia. PMID- 19304325 TI - 2-(1-Hydroxethyl)-4,8-dihydrobenzo[1,2-b:5,4-b']dithiophene-4,8-dione (BTP-11) enhances the ATRA-induced differentiation in human leukemia HL-60 cells. AB - 2-(1-Hydroxethyl)-4,8-dihydrobenzo[1,2-b:5,4-b']dithiophene-4,8-dione (BTP-11) is a potent enhancer for all-trans retinoic acid (ATRA)-induced differentiation in HL-60 cells. Combination of BTP-11 and ATRA cut down the concentration of ATRA significantly, and that BTP-11 promoted the progression of ATRA-induced into the terminal granulocytic differentiation. Further, Western blot analysis revealed that combination of BTP-11 and ATRA decreased cyclin D/CDK4 and increased C/EBPvarepsilon protein expression to arrest the cells into G0/G1 phase leading to granulocytic maturation. These results confirmed that BTP-11 is a potent enhancer for ATRA-induced differentiation of HL-60 cells, and the great developmental potential of BTP-11 will be expected. PMID- 19304326 TI - A measure of community members' perceptions of the impacts of research partnerships in health and social services. AB - Currently, there are no psychometrically sound outcome measures by which to assess the impacts of research partnerships. This article describes the development of a 33-item, survey questionnaire measuring community members' perceptions of the impact of research partnerships addressing health or social issues. The Community Impacts of Research Oriented Partnerships (CIROP) was developed using information from the literatures on health promotion, community development, research utilization, and community-based participatory research, and from focus groups involving 29 key informants. Data from 174 community members were used to determine the factor structure, internal consistency, and test-retest reliability of the four CIROP scales, and to provide evidence of construct validity. The CIROP informs research partnerships about the extent of their impact in the areas of Personal Knowledge Development, Personal Research Skill Development, Organizational/Group Access To and Use of Information, and Community and Organizational Development, allowing them to demonstrate accountability to funding bodies. As well, the CIROP can be used as a research tool to assess the effectiveness of knowledge sharing approaches, determine the most influential activities of research partnerships, and determine structural characteristics of partnerships associated with various types of impact. The CIROP provides a better understanding of community members' perspectives and expectations of research partnerships, with important implications for knowledge transfer and uptake. PMID- 19304327 TI - Immunohistochemical detection and localization of new type gosling viral enteritis virus in paraformaldehyde-fixed paraffin-embedded tissue. AB - To determine the distribution and localization of new type gosling viral enteritis virus (NGVEV) in paraformaldehyde-fixed paraffin-embedded tissues of experimentally infected goslings, for the first time, an immunohistochemical (IHC) staining method was reported. Anti-NGVEV polyclonal serum was obtained from the rabbits immunized with purified NGVEV antigen, which was extracted by caprylic-ammonium sulphate method and purified through High-Q columns anion exchange chromatography. Three-day-old NGVEV-free goslings were orally inoculated with NGVEV-CN strain suspension as infection group and phosphate buffered saline solution (PBS) as control group, respectively. The tissues were collected at sequential time points between 0.5 and 720h post inoculation (PI), and prepared for IHC staining and ultra-structural observation. The positive immunoreactivity could be readily detected in the lymphoid and gastrointestinal organs of infected goslings as early as 48 h PI, in the liver, kidney, pancreas and myocardium from 72 h, and in the cerebrum and cerebellum from 96 h, while it was hardly detected in the respiratory organs at any time. The positive staining reaction could be detected in NGVEV-infected goslings until 600 h PI, and no positive staining cell could be observed in the controls. The highest levels of viral antigen were found in the bursa of Fabricius (BF), thymus, proventriculus, gizzard and intestine tract, moreover, the liver, kidney, spleen, myocardium and pancreas were intensively and widely stained. The target cells had a ubiquitous distribution, especially included the epithelial cells, endothelial cells, superficial and crypt mucosal cells, glandular cells, fibrocytes, macrophages and lymphocytes, which served as the principal sites for antigen localization. The ultra structural observation by transmission electron microscope (TEM) further indicated that NGVEV particles could be widely detected in the lymphoid and digestive organs of infected goslings from 72 h PI onwards. This work may be useful not only for offering a possibility of routine diagnosis of NGVE, but also for better understanding of the pathogenesis of the disease. PMID- 19304328 TI - KIR2DL4 (CD158d) polymorphisms and susceptibility to multiple sclerosis. AB - Several lines of evidence implicate CD56(bright) NK cells in the pathogenesis of multiple sclerosis (MS). This proposed immunoregulatory pathway involves already established susceptibility genes such as interleukin-2 receptor alpha (IL2RA) and interleukin-7 receptor (IL7R). We therefore investigated the CD56(bright) NK cell effector molecule KIR2DL4 for its involvement in genetic susceptibility to MS in a study population of 763 cases and 967 controls. Whereas 26% of the study population has a genotype corresponding to a lack of any functional membrane bound form of the molecule, no association of the KIR2DL4 transmembrane alleles with susceptibility to MS was observed. PMID- 19304329 TI - Omics in algae: paving the way for a systems biological understanding of algal stress phenomena? AB - The last decade, the biological and biomedical scientific landscape has seen the increase in use and applications of "omics" technologies. These technologies provide methods that allow for a comprehensive description of nearly all components within the cell. Microalgae not only play an important ecological role, but are also of commercial importance and therefore call for an in depth knowledge of basic biological functions. Knowledge of separate algal subsystems has gradually become available, but the challenge remains to integrate data obtained from these subsystems and from different levels of biological organization. Systems biology is a discipline that aims at this integration. In this paper, the current status of "omics" in algae is reviewed. At the lowest level, genome studies and the use of microarrays seem to have found widespread acceptance in algal research. At higher level such as the proteome and metabolome, however, very few omic studies have been carried out in algae so far. Moreover, the need arises for the construction of computer databases to store obtained information in a systematic way. To illustrate the use and especially the future needs of algal "omics" in a systems biological context, a case study is presented in which a freshwater alga was subjected to heavy metal stress and toxicity endpoints were monitored on different levels of biological organization. PMID- 19304330 TI - Assessment of oxidative stress and histopathology in juvenile northern pike (Esox lucius) inhabiting lakes downstream of a uranium mill. AB - Lakes receiving effluent from the Key Lake uranium mill in northern Saskatchewan contain elevated trace metals, some of which are associated with increased reactive oxygen species (ROS) in cells and tissues causing oxidative stress. The potential for oxidative stress was assessed in juvenile (age 1+) northern pike (Esox lucius) collected from two exposure (high and low) and one reference lake near the Key Lake operation. The concentrations of total, reduced and oxidized glutathione and the ratio of oxidized to reduced glutathione in liver and kidney did not differ significantly among pike collected from exposure and reference lakes, with the exception of low exposure pike kidney that had significantly greater oxidized glutathione and ratio of oxidized to reduced glutathione. The concentrations of by-products of lipid peroxidation (malondialdehyde and 4 hydroxyalkenal) were significantly greater in kidney of pike collected from the reference lake compared to both exposure lakes. The activity of the antioxidant enzyme glutathione peroxidase in liver was greater in pike collected from the high exposure lake compared to the reference lake. Histopathological evaluations revealed greater pathology in reference lake pike as indicated by a greater number of pyknotic and fragmented nuclei and dilated tubules as well as a thickening of Bowman's capsule in kidney, and as a thickening of the primary filament epithelial padding in gills. In liver, hepatocyte morphology, including transsectional area and degree of vacuolation, differed among lakes without any clear signs of pathology. Trace metal analyses of muscle showed that eight elements (arsenic, cobalt, copper, iron, molybdenum, selenium, thallium, and uranium) were significantly elevated in pike collected from both exposure lakes compared to reference. These results provide only limited evidence of oxidative stress in exposure pike tissues and no evidence of histopathology despite indications that trace metals, most notably arsenic and selenium, were bioaccumulating in tissue. PMID- 19304331 TI - Elevated adiponectin levels in patients with chronic heart failure: an independent predictor of mortality or a marker of cardiac cachexia? AB - Adiponectin may influence the development of chronic heart failure (CHF), but the epidemiological data are scarce. This is due in part to the fact that while higher BMIs are a risk factor for CHF, obesity is a predictor of improved prognoses in patients with CHF since wasting is strongly associated with the increased risk of death in the final stages of CHF. From that standpoint of view, high adiponectin levels are a predictor of mortality in patients with CHF. That paradoxical relationship may exist since high BMI, hence low adiponectin, favors survival in endstage heart failure. We strongly believe that further large-scale clinical studies are warranted to analyze independent prognostic significance of adiponectin levels in patients with CHF. PMID- 19304332 TI - Factors associated with return to estrus in first service swine females. AB - Reproductive failure is a major factor contributing to the culling rate of swine females. As return to estrus is the most frequent reproductive failure, the study of factors associated with its occurrence can help to identify females with a predisposition to this failure. This study was run in a farm with a housing capacity of 1500 Agroceres PIC females in southern Brazil. Logistic regression models were run to determine the relative contribution of some factors to the probability of a sow returning to estrus. Return to estrus occurred in 27.8% (10/36) and 10.7% (50/469) of females remated after a previous reproductive failure and first service females, respectively. Females with a previous reproductive failure had 3.2-times higher odds (P=0.003) of returning to estrus than first service females. In first service females (n=469), higher odds (P<0.05) of returning to estrus were observed in parity order (PO) 0 (14.3%; 3.1 times), PO1 (20.6%; 4.9-times) and PO2 (12.7%; 2.7-times) compared to PO>2 (5.0%; reference class) females. Weaned females with ovarian cysts, detected during estrus, had 7.6-times higher (37.5%; 3/8) odds (P=0.012) of returning to estrus than females without cysts (8.3%; 28/338). In the model run with 330 weaned sows, the return to estrus rate was not affected (P>0.05) by the number of weaned piglets (7-9; 10; 11 and 12 piglets) but sows with a lactation length (LL) of 15 19 days had 3.5-times higher odds (P<0.05) of a return to estrus than sows with LL of 20-21 days. Return to estrus in weaned sows was also affected by the interaction between PO and body condition score (BCS) loss during lactation. Loss of more than 0.5 point in BCS resulted in higher odds (P<0.05) of returning to estrus in PO1 and PO2 but not in PO>2 sows. Accurate detection of estrus in gilts and adequate feed intake during lactation of first and second parity sows are suggested to reduce return to estrus rate in younger females. To avoid the detrimental effect of short lactations on reproductive performance options such as increasing the lactation length, split-weaning or skip-mating may be considered. PMID- 19304333 TI - Early changes of hepatitis B virus quasispecies during lamivudine treatment and the correlation with antiviral efficacy. AB - BACKGROUND/AIMS: To investigate dynamic changes of hepatitis B virus (HBV) quasispecies within the reverse transcriptase (RT) region during the early stage of lamivudine treatment and the correlation with antiviral efficacy. METHODS: Twenty-five chronic hepatitis B patients received lamivudine treatment for 48 weeks. Fourteen patients responded to lamivudine, while eleven patients were non responders. HBV DNA was extracted from serum samples at baseline and week 4. The RT region of HBV was amplified, then cloned and sequenced. Quasispecies complexity and diversity within the RT region were analyzed at baseline and week 4, and viral nucleotide substitution rates during the first 4 weeks were calculated. RESULTS: The quasispecies complexity and diversity were not different between responders and non-responders at baseline (p>0.05). However, the quasispecies complexity and diversity of responders were significantly lower than those of non-responders at week 4 (p<0.01). Furthermore, the viral nucleotide substitution rate of responders was significantly higher than that of non responders (p<0.05). CONCLUSIONS: The dynamic changes of HBV quasispecies within the RT region showed distinct patterns between responders and non-responders during early stage of lamivudine treatment. The dynamic changes of quasispecies complexity and diversity during the first 4 weeks were correlated with lamivudine antiviral efficacy and antiviral resistance. PMID- 19304334 TI - Selective ablation of Notch3 in HCC enhances doxorubicin's death promoting effect by a p53 dependent mechanism. AB - BACKGROUND/AIMS: The functional roles of endogenous Notch3 and Notch1 for protecting human hepatocellular carcinoma (HCC) lines against doxorubicin-induced death have been investigated. We previously reported aberrant Notch3 and Notch4 up-regulation in HCC and we have extended these observations to include Notch1. METHODS: Notch1 expression was assessed by immunohistochemistry and immunoblotting. Notch3 and Notch1 expression were ablated in multiple HCC lines by stable retroviral transduction of short hairpin RNAs (shRNAs). Effects on doxorubicin sensitivity were evaluated with respect to cell growth, expression of specific cell cycle effectors and multiple apoptotic parameters. RESULTS: Notch3 depletion increased p53 expression, doxorubicin uptake, DNA damage, the apoptosis inducing effects of doxorubicin and also impeded the cell cycle progression of HCC cells. Ablating p53 expression in Notch3 knockdown (KD) cells largely abolished their enhanced doxorubicin sensitivity; and Notch3 KD in p53(-/-) Hep3B cells failed to influence their response to doxorubicin. Although up-regulated in most HCC, Notch1 (unlike Notch3) did not contribute to the doxorubicin resistance of HCC lines. CONCLUSIONS: Our in vitro results represent the first evidence that Notch3 silencing in combination with chemotherapeutics could conceivably provide a novel strategy for HCC treatment that deserves further exploration. PMID- 19304335 TI - Low serum concentration of apolipoprotein A-I is an indicator of poor prognosis in cirrhotic patients with severe sepsis. AB - BACKGROUND/AIMS: Severe sepsis is frequently associated with hypocholesterolemia which is also a common finding in cirrhotic patients. Lipoprotein is capable of binding endotoxin to which cirrhotic patients exhibit an excessive pro inflammatory response. METHODS: We evaluated the relationship between lipid levels, inflammatory cytokines and clinical outcomes in 103 cirrhotic patients with severe sepsis. RESULTS: The non-survivors had significantly lower concentrations of total cholesterol, high-density lipoprotein (HDL), and apolipoprotein A-I (APO A-I). HDL and APO A-I levels were inversely correlated with interleukin-6, tumor necrosis factor-alpha, and various disease severity scores. Serum creatinine, mean arterial pressure and low level of APO A-I (<47.5mg/dl) were independent factors to predict 90-day mortality. The cumulative survival rates at 90 days were 63.8% and 8.9% for the high APO A-I and low APO A I groups (p<0.0001). Low APO A-I was also associated with lower mean arterial pressure, higher rate of vasopressor dependency, and greater plasma renin activity. CONCLUSIONS: Serum levels of HDL and APO A-I are inversely correlated with liver reserve and disease severity in cirrhotic patients with severe sepsis. Low level of APO A-I is associated with a marked impairment of effective arterial volume, multiple organ dysfunction and a poor prognosis. PMID- 19304336 TI - High levels of factor VIII and risk of extra-hepatic portal vein obstruction. AB - BACKGROUND/AIMS: High levels of coagulation factor VIII are a risk factor for lower-limb deep vein thrombosis (DVT). Their role in extra-hepatic portal vein obstruction (EHPVO) is not established. METHODS: Factor VIII was measured in 85 patients with EHPVO (primary in 58 and complicating liver cirrhosis in 27), in 200 with lower-limb DVT, in 108 with liver cirrhosis without thrombosis and in 200 healthy controls. RESULTS: Factor VIII levels were significantly higher in patients with primary EHPVO (138 IU/dL, range 86-366), EHPVO and cirrhosis (147 IU/dL, 95-242), lower-limb DVT (140 IU/dL, 64-400) and cirrhosis alone (160 IU/dL, 43-446) than in controls (112 IU/dL, 62-250, p<0.001). When factor VIII exceeded 129 IU/dL (66th percentile), the odds ratios were 10.5 (95%CI 3.3-33.4) for primary EHPVO, 6.0 (1.2-30.7) for EHPVO and cirrhosis, 5.0 (2.6-9.4) for lower-limb DVT. After exclusion of the effect of the acute phase reaction, the odds ratio for primary EHPVO was 4.2 (0.8-22.7), and was 8.7 (0.9-80.5) after exclusion also of patients with chronic myeloproliferative disorders. CONCLUSIONS: High factor VIII levels are independently associated with an increased risk for EHPVO. The risk of EHPVO increased with increasing factor VIII levels and was only partially dependent on the acute phase reaction. PMID- 19304337 TI - Capacity to control noncommunicable diseases in the countries of South-East Asia. AB - OBJECTIVE: To assess the status of national capacity for prevention the and control of noncommunicable diseases (NCDs) in the Member States of the South-East Asia (SEA) Region of the World Health Organization (WHO). METHODS: A survey was conducted in 2006-2007 in all 11 Member States of the SEA Region of WHO using a regional adaptation of the global WHO questionnaire. Information was collected on the existence of national legislation, policies, strategies, and programmes, implementation status of the WHO Framework Convention on Tobacco Control, and the Global Strategy on Diet, Physical Activity and Health, and the availability of services and financial resources for the prevention and control of NCDs. The results of this survey were compared with those of a similar survey conducted in the region in 2001. RESULTS: The comparison of two surveys showed progress made in the Region between 2001 and 2006 in strengthening commitment and capacity to prevent and control NCDs. Major gaps remain in the creation of appropriate legislative and regulatory environment, policy formulation for the promotion of healthy nutrition and physical activity, and generation of guidelines for the management of NCDs and their risk factors. CONCLUSION: Member States of the SEA Region need to enhance the capacity to address critical gaps in their national polices and programmes for the prevention and control of NCDs. This requires technical support of WHO and other developmental partners. PMID- 19304338 TI - FOXP2 as a molecular window into speech and language. AB - Rare mutations of the FOXP2 transcription factor gene cause a monogenic syndrome characterized by impaired speech development and linguistic deficits. Recent genomic investigations indicate that its downstream neural targets make broader impacts on common language impairments, bridging clinically distinct disorders. Moreover, the striking conservation of both FoxP2 sequence and neural expression in different vertebrates facilitates the use of animal models to study ancestral pathways that have been recruited towards human speech and language. Intriguingly, reduced FoxP2 dosage yields abnormal synaptic plasticity and impaired motor-skill learning in mice, and disrupts vocal learning in songbirds. Converging data indicate that Foxp2 is important for modulating the plasticity of relevant neural circuits. This body of research represents the first functional genetic forays into neural mechanisms contributing to human spoken language. PMID- 19304340 TI - Polymorphisms of the ribonucleotide reductase M1 gene and sensitivity to platin based chemotherapy in non-small cell lung cancer. AB - Ribonucleotide reductase catalyzes the rate limiting step of deoxyribonucleotide formation, a crucially important step in DNA synthesis and repair. The regulatory subunit M1 of ribonucleotide reductase (RRM1) is the necessary part of the RR function and controls substrate specificity and global on/off enzyme activity. Despite recent research progress, the role of RRM1 in lung cancer sensitivity to chemotherapeutics remains to be elucidated. This study was to investigate the relationship between polymorphisms of the RRM1 gene and sensitivity to platinum based chemotherapy in non-small cell lung cancer (NSCLC). Genomic DNA samples from 214 NSCLC patients treated with platinum-based chemotherapy were used to determine the RRM1 promoter allelotypes. The RR37CC-RR524TT was the most frequent allelotype (38.50%), followed by RR37AC-RR524CT (26.76%) and RR37CC-RR524CT (14.95%). The average response rate for chemotherapy was 44.4%. The response rates to the treatment regimens in the RR37CC-RR524TT, RR37AC-RR524CT and RR37CC RR524CT allelotypes were 43.9%, 52.6%, and 51.6%, respectively. The response rates to therapy among patients with RRM1 (-)524 allelotypes were significantly different (p=0.046), whereas that among patients with RRM1 (-)37 allelotypes were not significant. Further analysis showed that the response rate in the patients with RR524CT allelotype (52.3%) was the highest, compared with that with RR37CC RR524TT allelotype (43.9%, p=0.28), or the Others (RR524CC and RR37AC-RR524TT, 30.2%, p=0.02). Our results suggest that the RR524CT allelotype may be associated with an increased sensitivity to platinum-based chemotherapy in NSCLC. Further research on determining RR524CT as a clinical marker for predicting response to platinum-based therapy in NSCLC patients is warranted. PMID- 19304339 TI - Impact and interactions between smoking and traditional prognostic factors in lung cancer progression. AB - BACKGROUND: Cigarette smoking is a well-known risk factor of lung carcinogenesis. The clinical impact of smoking on lung cancer metastases and survival remains unclear. We sought to investigate the effect of smoking intensity on lung cancer treatment failure (represented by overall survival), and the interactions between smoking and clinicopathological factors in lung cancer progression. METHODS: Clinical information was obtained from four non-small cell lung cancer patient cohorts (n=347). Twenty patients were excluded from the analysis because their smoking history was not available. The distribution of smoking intensity on patient age (> or =60 years or <60 years), gender, tumor differentiation (poor, moderate and well differentiated), and clinical stage (1, 2, or 3) was assessed with Kruskal-Wallis rank sum tests. The effect of smoking on cause-specific lung cancer mortality was estimated by using Cox proportional hazard models and Kaplan Meier analysis. The interactions between clinicopathological factors and smoking intensity with regard to lung cancer overall survival were evaluated with analysis of variance (ANOVA) for Cox modeling. RESULTS: Greater smoking intensity at diagnosis was found in older patients (> or =60 years; p=0.022), male (p=1.35e 7), poorly differentiated tumors (p=8.51e-5), patients with tumor stage 2 (p=0.031), and squamous cell lung cancer patients (p=2.2e-16). Patients who smoked more than 61 packs/year had an increased risk for lung cancer recurrence (hazard ratio=1.41, 95% CI: [1.03, 1.94], log-rank p=0.032) and shorter overall survival period (log-rank p=0.033, Kaplan-Meier analysis) than those who smoked less than 61 packs/year. ANOVA analysis showed that smoking intensity (p=0.03) and tumor stage (p=1.2e-6) are the only significant prognostic factors of lung cancer, whereas patient age, gender, and tumor differentiation were not significant in lung cancer prognostication. There were significant interactions between smoking and clinical stage (p=0.02) as well as patient age and tumor differentiation (p=0.03) in lung cancer progression. CONCLUSION: Smoking intensity at diagnosis is an independent, significant prognostic factor of non small cell lung cancer. This factor could be used in patient selection for chemoprevention of tumor metastases and relapse. Additionally, the information may be used for clinically relevant tobacco prevention and intervention messages. PMID- 19304341 TI - Protein 4.1 and its interaction with other cytoskeletal proteins in Xenopus laevis oogenesis. AB - In human red blood cells, protein 4.1 (4.1R) is an 80-kDa polypeptide that stabilizes the spectrin-actin network and anchors it to the plasma membrane. In non-erythroid cells there is a great variety of 4.1R isoforms, mainly generated by alternative pre-mRNA splicing, which localize at various intracellular sites, including the nucleus. We studied protein 4.1R distribution in relation to beta spectrin, actin and cytokeratin during Xenopus oogenesis. Immunoprecipitation experiments indicate that at least two isoforms of protein 4.1R are present in Xenopus laevis oocytes: a 56-kDa form in the cytoplasm and a 37-kDa form in the germinal vesicle (GV). Antibodies to beta-spectrin reveal two bands of 239 and 100 kDa in the cytoplasm. Coimmunoprecipitation experiments indicate that both the 37- and 56-kDa isoforms of protein 4.1R associate with the 100-kDa isoform of beta-spectrin. Moreover, the 56-kDa form coimmunoprecipitates with a cytokeratin of the same molecular weight. Confocal immunolocalization shows that protein 4.1R distribution is in the peripheral cytoplasm, in the mitochondrial cloud (MC) and in the GV of previtellogenic oocytes. In the cytoplasm of vitellogenic oocytes, a loose network of fibers stained by the anti-protein 4.1R antibody spreads across the cytoplasm. beta-Spectrin has a similar distribution. Protein 4.1R was found to colocalize with actin in the cortex of oocytes in the form of fluorescent dots. Double immunolocalization of protein 4.1R and cytokeratin depicts two separate networks that overlap throughout the whole cytoplasm. Protein 4.1R filaments partially colocalize with cytokeratin in both the animal and vegetal hemispheres. We hypothesize that protein 4.1R could function as a linker protein between cytokeratin and the actin-based cytoskeleton. PMID- 19304342 TI - Zn stress-induced inhibition of bean PvSR2-GUS fusion gene splicing is gene specific in transgenic tobacco. AB - The stress-related gene no. 2 of Phaseolus vulgaris (PvSR2) is metal inducible and contains a single intron. Here, we report that Zn stress inhibited the splicing of the PvSR2-beta-glucuronidase (GUS) fusion gene in a concentration- and time-dependent manner in tobacco seedlings. The inhibition appears to be specific for the PvSR2-GUS transgene: splicing of four endogenous tobacco genes was unaffected by Zn stress. Our results provide in vivo evidence that Zn stress dependent intron retention is transgene specific in plants. PMID- 19304343 TI - Evidence-based emergency medicine/editorial. Contrasting agendas: science for the bedside or water in the wind? PMID- 19304344 TI - Transport of patients after out-of-hospital cardiac arrest: closest facility or most appropriate facility? PMID- 19304345 TI - [Heart failure and geriatrics]. PMID- 19304346 TI - [semFYC therapeutic guidelines in primary care. Essential Arterial Hypertension Section: Updated? Evidence based?]. PMID- 19304347 TI - [Clinical competence of family medicine medical residents in the most common skin diseases]. PMID- 19304348 TI - [Prevention of colorectal cancer]. PMID- 19304349 TI - [Detection and management of Trypanosoma cruzi in primary care in non-endemic areas]. PMID- 19304350 TI - [Quinolone-induced rupture of the long extensor tendon of the thumb]. PMID- 19304351 TI - [semFYC and ezetimib]. PMID- 19304352 TI - [Telemedicine and primary care]. PMID- 19304353 TI - [Policies to diminish the inequality in health in Spain: a few precisions]. PMID- 19304354 TI - Potential of mean force for Syrian hamster prion epitope protein--monoclonal fab 3f4 antibody interaction studies. AB - Simulating antigen-antibody interactions are crucial for understanding antigen antibody associations in immunology. To shed further light on this question, we study a dissociation of the Syrian hamster prion epitope protein-fab 3f4 antibody complex structure. The stretching, that is, the distance between the center of mass of the prion epitope protein and the fab 3f4 antibody, has been studied using potential of mean force (PMF) calculations based on molecular dynamics (MD) and the implicit water model. For the complex structure, there are four important intermediates, U-shaped groove on the antibodies, and two inter-protein molecular hydrogen bonds in the stretching process. Use of our simulations may help in understanding the binding mechanics of the complex structure, and thus of significance in the design of antibodies against prion disease. PMID- 19304355 TI - [Abnormal coronarography]. PMID- 19304356 TI - [Hypoplasia of the hepatic segment of the inferior vena cava]. AB - Congenital anomalies of the inferior vena cava are not uncommon. In the past, the diagnosis was made by angiography and retrograde right heart studies which were being commonly performed for the assessment of congenital malformation of the heart. The advent of ultrasound and three-dimensional imaging such as computed tomography and magnetic resonance made it easier to obtain the diagnosis. We report here a case of inferior vena cava hypoplasia in a 19-year-old male, who presented with progressive ascitis and edema. PMID- 19304357 TI - [Photosensitivity in lupus erythematosus]. AB - Photosensitivity is one of the ARA diagnostic criteria of systemic lupus erythematosus. Sun exposure can also induce extracutaneous manifestations of the disease. Photosensitivity may be difficult to prove by history taking in lupus patients, as the delay between sun exposure and the onset of specific skin lesions is rather long. Photo-induction of lupus can occur by ultraviolet A (UVA) radiation in the shadow or behind window glass, so that the relationship between radiation exposure and exacerbation of the disease may not seem obvious to the patient. Phototesting procedures for lupus erythematosus have been described, but they are not used in routine practice. Both UVB and UVA play a role in the pathogenesis of lupus erythematosus: in the epidermis they induce DNA damage, they expose nuclear antigens and photo-induced neo-antigens at the cell surface, they lead to an accumulation of apoptotic material, and they induce several pro inflammatory cytokines. In the dermis, UV radiation triggers skin infiltration by inflammatory cells by modulation of microvascular flow rates and by upregulation of white blood cell migration from dermal capillaries to the skin. Photodistribution of skin lesions and a delay of their onset of more than 48 hours after sun exposure are clinical hallmarks of cutaneous lupus erythematosus that are usually completed by histological confirmation. Photoprotection is essential in the treatment of lupus patients: it comprises sun avoidance suitable for both UVB and UVA radiation, protective clothing, and topical broad-spectrum filters. PMID- 19304358 TI - [Neurogenic osteoarthropathy of the knee associated with spina bifida: a diagnosis not to be missed]. AB - Neurogenic arthropathy is a severe complication of chronic sensitive deficits that occurred commonly in diabetic neuropathies. It is a destructive and painless osteoarthritis associated with a loss of the deep sensitivity and a defect of protective reactions against chronic articular microtraumatisms. We report a 55 year-old woman with neuroarthropathy of the knee resulting from a spina bifida. Bisphosphonate use is an effective but non-consensual treatment. PMID- 19304359 TI - Sources and pathways of 90Sr in the North Atlantic-Arctic region: present day and global warming. AB - The spatial and temporal distributions of the anthropogenic radionuclides (137)Cs and (90)Sr, originating from nuclear bomb testing, the Sellafield reprocessing plant in the Irish Sea (UK), and from the Ob and Yenisey river discharges to the Arctic Ocean, have been simulated using the global version of the Miami Isopycnic Coordinate Ocean Model (MICOM). The physical model is forced with daily atmospheric re-analysis fields for the period of 1948-1999. Comparison of the temporal evolution of the observed and the simulated concentrations of (90)Sr has been performed in the Kara Sea. The relative contributions of the different sources on the temporal and spatial distributions of the surface (90)Sr are quantified over the simulated period. It follows that the Ob river discharge dominated the surface (90)Sr over most of the Arctic Ocean and along the eastern and western coasts of Greenland before 1960. During the period of 1980-1990, the atmospheric fallout and the Ob river discharge were equally important for the (90)Sr distribution in the Arctic Ocean. Furthermore, an attempt has been made to explore the possible dispersion of accidental released (90)Sr from the Ob and Yenisey rivers under a global warming scenario (2 x CO(2)). The difference between the present-day and the global warming scenario runs indicates that more of the released (90)Sr from the Ob and Yenisey rivers is confined to the Arctic Ocean in the global warming run, particularly in the near coastal, non-European part of the Arctic Ocean. PMID- 19304360 TI - Mercury contamination in vicinity of secondary copper smelters in Fuyang, Zhejiang Province, China: levels and contamination in topsoils. AB - In the present study, we aim to investigate the extent of soil contamination by Hg, particularly by anthropogenic Hg, and tentatively estimate the total Hg (Hg(T)) accumulation in topsoils (0-15 cm) in Fuyang, Zhejiang Province--a secondary Cu smelter of China. The results show that the levels of soil Hg in the vicinity of the smelters have been substantially elevated following local smelting activities. The spatial distribution of soil Hg in this area reveals a rapid decrease as the distance from the smelter reaches 1.5 km, which is probably due to the quick deposition process of particulate Hg and reactive gaseous Hg emitted from the smelters. The total accumulation of Hg(T) in the topsoils of the study area of 10.9 km(2) is approximately 365-561 kg and of which 346-543 kg might be contributed by anthropogenic emission alone with an annual emission of 17.3-27.2 kg Hg to the topsoils. PMID- 19304361 TI - Testing the influenza-tuberculosis selective mortality hypothesis with Union Army data. AB - Using Cox regression, this paper shows a weak association between having tuberculosis and dying from influenza among Union Army veterans in late nineteenth-century America. It has been suggested elsewhere [Noymer, A. and M. Garenne (2000). The 1918 influenza epidemic's effects on sex differentials in mortality in the United States. Population and Development Review 26(3), 565 581.] that the 1918 influenza pandemic accelerated the decline of tuberculosis, by killing many people with tuberculosis. The question remains whether individuals with tuberculosis were at greater risk of influenza death, or if the 1918/post-1918 phenomenon arose from the sheer number of deaths in the influenza pandemic. The present findings, from microdata, cautiously point toward an explanation of Noymer and Garenne's selection effect in terms of age-overlap of the 1918 pandemic mortality and tuberculosis morbidity, a phenomenon I term "passive selection". Another way to think of this is selection at the cohort, as opposed to individual, level. PMID- 19304362 TI - [Health-care quality standards in chronic obstructive pulmonary disease]. AB - Although clinical practice guidelines have contributed to improving the quality of health care offered to patients with chronic obstructive pulmonary disease (COPD), the level of adherence to recommendations continues to be inadequate and variable. Standards of care in COPD are written after applying an evidence-based approach, with the aim of unifying health-care criteria, establishing levels of acceptable adherence, and providing a way to assess quality; the ultimate goal is to improve patient care. In this statement we propose a series of health-care quality criteria and related indicators that will facilitate the quantitative evaluation of adherence to recommendations. The level of adherence that should be required is stipulated. This statement is not intended to provide a detailed description of how COPD should be managed. The aim is rather to set out quality assurance criteria that will contribute to the improvement of health-care access and equity, guaranteeing application of the highest levels of scientific and technical quality possible within the constraints of available resources, with the final purpose of satisfying the patient with COPD. The quality criteria have been grouped in 3 categories: a) so-called key criteria, to which adherence is essential; b) a set of conventional quality standards; and c) health-care administrative standards. Finally, we propose a framework on which to base the eventual accreditation of health-care quality for COPD patients. PMID- 19304363 TI - [Usefulness of the N-terminal fraction of brain natriuretic peptide for deciding when to refer patients with sleep apnea-hypopnea syndrome to the cardiologist]. AB - INTRODUCTION AND OBJECTIVE: When sleep apnea-hypopnea syndrome (SAHS) and cardiovascular disease occur concurrently, prognosis is affected. Echocardiography can detect structural cardiac abnormalities but using this technique in all patients would place a heavy burden on resources. The objective of this study was to investigate whether the N-terminal fraction of brain natriuretic peptide (NT-proBNP) can be used as a marker for silent heart disease. PATIENTS AND METHODS: NT-proBNP concentration was measured in the 114 consecutive patients with SAHS who underwent echocardiography before starting treatment. Left and right ventricular systolic and diastolic function, as well as structural abnormalities, were studied. Correlations between NT-proBNP concentration and the abnormalities detected were investigated. A receiver operating characteristics (ROC) curve was plotted for NT-proBNP concentration and cardiac abnormalities. RESULTS: Data for 98 patients were finally analyzed. NT-proBNP concentration was significantly correlated with ventricular septal thickness (r=0.63), posterior wall thickness (r=0.45), and left ventricular end-diastolic diameter (r=0.51) (P<.0001 for all correlations). The area under the ROC curve was significant (0.870; 95% confidence interval, 0.801-0.939; P<.0001). Assuming that specificity would be more useful for clinical practice, we calculated that NT-proBNP concentrations below 100 and 200 pg/mL could rule out structural abnormalities with a reliability of 90% and 100%, respectively. CONCLUSIONS: NT-proBNP concentration was strongly correlated with echocardiographic abnormalities and so could be a useful tool for identifying patients who should be referred to the cardiologist. PMID- 19304364 TI - Postpartum toxic shock syndrome due to methicillin-resistant Staphylococcus aureus epidemic in community. PMID- 19304365 TI - Presealing of the chorioamniotic membranes prior to fetoscopic surgery: preliminary study with unfertilized chicken egg models. AB - BACKGROUND: Fetoscopic surgical techniques continue to develop. However, progress has been hindered by premature rupture of membranes (PROM), which complicates 5 30% of fetoscopic procedures. Several membrane closure techniques have been devised but none proven reliable. OBJECTIVE: We propose a new approach that of presealing the chorioamniotic membrane prior to membrane disruption-a so called Amnioseal. A set of pilot experiments were designed using unfertilized chicken egg models to test our proposal. STUDY DESIGN: Two novel unfertilized chicken egg models were developed. Model 1 simulated the chorioamniotic membrane and amniotic cavity. Model 2 simulated the uterine muscle/chorioamniotic membrane interface and amniotic cavity. Four sealants (100% petroleum jelly, FloSeal Hemostatic Matrix, CoSeal Hemostatic Matrix and BioGlue Surgical Adhesive) were tested against untreated controls. The sealants were applied directly to the egg membranes followed by biopsy needle puncture and needle membrane manipulation. RESULTS: BioGlue adhered strongly to the membrane correlating with the smallest defect size, greatest resistance to rupture, lowest degree of leakage and formed a water tight seal around the needle during membrane manipulation. In comparison, the weak adherence of FloSeal correlated with a larger defect size and higher degree of leakage. 100% petroleum jelly was non-adhesive, provided no membrane support and resulted in membrane rupture. CONCLUSION: Adhesive sealants confer mechanical support to the membrane and form a water tight seal. Experiments show that Sealant properties greatly affect outcomes. As such the Amnioseal's success will be determined by the properties of the sealant used. Specifically designed sealants are being developed along side a delivery device and will be tested using an in vitro human chorioamniotic membrane model. PMID- 19304366 TI - Fetal size and growth velocity in the prediction of shoulder dystocia in a glucose impaired population. PMID- 19304367 TI - Bioengineering aspects of the umbilical cord. AB - The umbilical cord and its constituent tissues: an outer layer of amnion, porous Wharton's jelly, two umbilical arteries, and one umbilical vein, are designed to protect blood flow to the fetus during a term pregnancy. The outer amnion layer may regulate fluid pressure within the umbilical cord. The porous, fluid filled Wharton's jelly likely acts to prevent compression of the vessels. Blood flow is regulated by smooth muscle surrounding the arteries that is intermingled with a collagen based extracellular matrix (ECM). Doppler ultrasound measurements of blood flow within the umbilical cord, and at specific sites within the developing fetus, provide evidence of impaired blood flow in conditions such as preeclampsia. Mechanosensory communication between cells and the extracellular matrix (ECM) may likely result in cords possessing abnormal physical dimensions, impaired hemodynamics, and altered composition within the umbilical cord tissues. Few studies have explored the biomechanics of the intact umbilical cord, with its constituent tissues, from normal pregnancies or abnormal pregnancies, maternal or fetal complications. Here, alterations in the umbilical cord are reviewed concerning anatomical abnormalities, disease, or chromosomal alterations using sonography, Doppler ultrasound, histology, and biomolecular and biochemical analyses. This paper considers how current knowledge of the umbilical cord and its constituent tissues can be used to infer biomechanical function. In addition, the mechanical consequences of structural abnormalities and altered tissue structure or composition are discussed with a specific focus on preeclampsia. PMID- 19304368 TI - Synthesis of calcium phosphate hydrogel from waste incineration fly ash and its application to fuel cell. AB - Waste incineration fly ash was successfully recycled to calcium phosphate hydrogel, a type of fast proton conductor. The crystallized hydrogel from incineration fly ash had a lower electric conductivity and a lower crystallinity than that from calcium carbonate reagent. However, the difference in electric conductivity between these crystallized hydrogels decreases with temperature. This was due to the presence of potassium in the incineration fly ash. The fuel cell with a membrane electrode assembly (MEA) using the calcium phosphate hydrogel membrane prepared from incineration fly ash was observed to generate electricity. The performance of this fuel cell was almost equal to that of a mixture of K(2)CO(3) and CaCO(3) reagents; further, the performance of the former was superior to the fuel cell with a perfluorosulfonic polymer membrane at temperatures greater than approximately 85 degrees C. PMID- 19304369 TI - Editorial comment on: Tension-free vaginal tape in the management of recurrent urodynamic stress incontinence after previous failed midurethral tape. PMID- 19304370 TI - Tension-free vaginal tape in the management of recurrent urodynamic stress incontinence after previous failed midurethral tape. AB - BACKGROUND: Data about the use of tension-free vaginal tape (TVT) in the management of recurrent urodynamic stress incontinence (RUSI) after previous failed midurethral sling procedure (MUSP) are limited. OBJECTIVE: Assessment of the efficacy and the indications of the TVT procedure in the management of patients with RUSI after failed previous MUSP. DESIGN, SETTING, AND PARTICIPANTS: Thirty-one patients with RUSI after previous failed MUSP were prospectively enrolled at a single tertiary academic center. MEASUREMENTS: Preoperatively and postoperatively, patients were assessed with physical examination, urinalysis, urine culture, bladder diary for 2-3 d, Q-tip test, uroflow, filling and voiding cystometry, urethral profilometry, and 1-h pad test. Mean follow-up was at 18.6 mo (range: 12-28 mo). RESULTS AND LIMITATIONS: Overall, the objective cure rate based on the pad test findings was 74%, the improvement rate was 6.5%, and the failure rate was 19.5%. The objective cure rate based on cough stress test during filling cystometry was 77.4%, and the subjective cure rate based on patients' answers was 71%. The study could have some limitations. The relatively small number of patients enrolled could affect the findings of study to some degree. Additionally, because urethral pressure profiles show a significant degree of directional dependence when side-hole microtip transducers are used, as in the present study, the orientation of the transducer could affect the values measured. CONCLUSIONS: The TVT procedure as a second operation could provide an overall cure rate of 74% with a low complication rate in female patients with RUSI after previous failed midurethral tape procedures. PMID- 19304371 TI - Editorial comment on: Tension-free vaginal tape in the management of recurrent urodynamic stress incontinence after previous failed midurethral tape. PMID- 19304372 TI - Prostate cancer gene 3 (PCA3): development and internal validation of a novel biopsy nomogram. AB - BACKGROUND: Urinary prostate cancer gene 3 (PCA3) represents a promising novel marker of prostate cancer detection. OBJECTIVE: To test whether urinary PCA3 assay improves prostate cancer (PCa) risk assessment and to construct a decision making aid in a multi-institutional cohort with pre-prostate biopsy data. DESIGN, SETTING, AND PARTICIPANTS: PCA3 assay cut-off threshold analyses were followed by logistic regression models which used established predictors to assess PCa-risk at biopsy in a large multi-institutional data set of 809 men at risk of harboring PCa. MEASUREMENTS: Regression coefficients were used to construct four sets of nomograms. Predictive accuracy (PA) estimates of biopsy outcome predictions were quantified using the area under the curve of the receiver operator characteristic analysis in models with and without PCA3. Bootstrap resamples were used for internal validation and to reduce overfit bias. The extent of overestimation or underestimation of the observed PCa rate at biopsy was explored graphically using nonparametric loss-calibration plots. Differences in PA were tested using the Mantel-Haenszel test. Finally, nomogram-derived probability cut-offs were tested to assess the ability to identify patients with or without PCa. RESULTS AND LIMITATIONS: PCA3 was identified as a statistically independent risk factor of PCa at biopsy. Addition of a PCA3 assay improved bootstrap-corrected multivariate PA of the base model between 2% and 5%. The highest increment in PA resulted from a PCA3 assay cut-off threshold of 17, where a 5% gain in PA (from 0.68 to 0.73, p=0.04) was recorded. Nomogram probability-derived risk cut-off analyses further corroborate the superiority of the PCA3 nomogram over the base model. CONCLUSIONS: PCA3 fulfills the criteria for a novel marker capable of increasing PA of multivariate biopsy models. This novel PCA3-based nomogram better identifies men at risk of harboring PCa and assists in deciding whether further evaluation is necessary. PMID- 19304373 TI - Editorial comment on: Tension-free vaginal tape in the management of recurrent urodynamic stress incontinence after previous failed midurethral tape. PMID- 19304374 TI - Editorial comment on: Prostate cancer gene 3 (PCA3): development and internal validation of a novel biopsy nomogram. PMID- 19304375 TI - Identification of tumor-associated antigens by using SEREX in hepatocellular carcinoma. AB - AIM: To identify biomarkers for diagnosis and prognosis of hepatocellular carcinoma (HCC). METHODS: Screening the HCC cDNA library with HCC patients sera. Isolated proteins were used as antigens to detect antibodies from patients with HCC and control sera. RESULTS: Eighty-one positive clones were identified. The frequencies of autoantibody against five HCC-associated antigens were higher in HCC than that in chronic hepatitis and normal human sera. The sensitivity and specificity of KRT23, AHSG and FTL antigens combination tests up to 98.2% in joint test and 90.0% in series test separately. CONCLUSIONS: HCC associate antigens identified from this study supply candidate markers of diagnosis, combined detection and immunotherapy of HCC. PMID- 19304376 TI - Adsorption characteristics of N-nitrosodimethylamine from aqueous solution on surface-modified activated carbons. AB - This study investigated the removal of N-nitrosodimethylamine (NDMA) by an adsorption mechanism using commercially available activated carbons and surface modified activated carbons. The effects of the modification on the properties of the activated carbon were studied by N(2) adsorption/desorption, Diffuse Reflectance Infrared Fourier Transmission (DRIFT) analysis and X-Ray Photoelectron Spectroscopy (XPS). Adsorption experiments revealed that the activated carbons demonstrated a greater capacity for NDMA adsorption capacity than can be achieved using zeolite. The equilibrium data was fitted to the Freundlich equation and it was found that the adsorption capacity was significantly influenced by the micropore size, relative pore volume and surface characteristics. Adsorption experiments were conducted using unmodified and modified activated carbons. The results indicated that the adsorption capacity of NDMA can be significantly improved by heat treatment and doping of TiO(2) particles. This was because the surface treatments yielded more hydrophobic sites and fewer oxygen-containing surface functional groups, and consequently an increased capacity for NDMA adsorption. PMID- 19304377 TI - Removal of ammonia nitrogen in wastewater by microwave radiation: a pilot-scale study. AB - A large removal of ammonia nitrogen in wastewater has been achieved by microwave (MW) radiation in our previous bench-scale study. This study developed a continuous pilot-scale MW system to remove ammonia nitrogen in real wastewater. A typical high concentration of ammonia nitrogen contaminated wastewater, the coke plant wastewater from a Coke company, was treated. The output power of the microwave reactor was 4.8 kW and the handling capacity of the reactor was about 5m(3) per day. The ammonia removal efficiencies under four operating conditions, including ambient temperature, wastewater flow rate, aeration conditions and initial concentration were evaluated in the pilot-scale experiments. The ammonia removal could reach about 80% for the real coke-plant wastewater with ammonia nitrogen concentrations of 2400-11000 mg/L. The running cost of the MW technique was a little lower than the conventional steam-stripping method. The continuous microwave system showed the potential as an effective method for ammonia nitrogen removal in coke-plant water treatment. It is proposed that this process is suitable for the treatment of toxic wastewater containing high concentrations of ammonia nitrogen. PMID- 19304378 TI - Effects of organoclays on soil eubacterial community assessed by molecular approaches. AB - The aim of this study was to evaluate the effects of the commercial organoclays, CLOISITE 30B, NANOFIL 804 and DELLITE 26C on soil eubacterial community. An enrichment test was carried out on Nutrient Broth containing the organoclay and the microorganisms previously isolated from soil. Four transfers were made, each after 7 days incubation. The molecular analyses on the eubacterial community were performed before treatment and 7 days after each transfer. DNA was extracted, amplified with eubacterial primers, finally analysed by amplified ribosomal DNA restriction analysis (ARDRA) and denaturing gradient gel electrophoresis (DGGE). The profiles of the samples treated with each organoclay showed the absence, the appearance and an increase in the intensity of some bands. These bands were excised from the gels, and the related microorganisms were identified by DNA sequencing, as Pseudomonas putida, Alcaligenes xylosoxidans, Pseudomonas monteilii and Pseudomonas aeruginosa. NAN804 treatment did not have any influence on soil eubacterial community, CLO30B had a slight toxic effect only on P. putida, instead the DEL26C treatment had a stronger toxic effect on P. putida and a slight toxic effect on P. monteilii. Finally, all the tested organoclays stimulated the growth of both A. xylosoxidans and P. aeruginosa. PMID- 19304379 TI - Accumulation of arsenic and nutrients by castor bean plants grown on an As enriched nutrient solution. AB - Phytoextraction is a remediation technique that consists in using plants to remove contaminants from soils and water. This study evaluated arsenic (As) accumulation in Castor bean (Ricinus communis cv. Guarany) grown in nutrient solution in order to assess its phytoextraction ability. Castor bean plants were grown under greenhouse conditions in pots containing a nutrient solution amended with increasing doses of As (0, 10, 50, 100, 250, 500 and 5000 microg L(-1)) in a completely randomized design with four replications. Shoot and roots dry matter production as well as arsenic and nutrient tissue concentrations were measured at the end of the experiment. The results showed that increasing As concentration in nutrient solution caused a decrease in shoot and root biomass but did not result in severe toxicity symptoms in castor bean growing under a range of As concentration from 0 to 5000 microg L(-1). The As doses tested did not affect the accumulation of nutrients by castor bean. Although castor bean did not pose characteristics of a plant suitable for commercial phytoextraction, it could be useful for revegetation of As-contaminated areas while providing an additional income by oil production. PMID- 19304380 TI - Kinetics of pulp mill effluent treatment by ozone-based processes. AB - The wastewaters generated from wood pulping and paper production processes are traditionally treated by biological and physicochemical processes. In order to reduce chemical oxygen demand (COD) and color to meet increasingly strict discharge standards, advanced oxidation processes (AOPs) are being adapted as polishing treatment units. Various ozone-based processes were used in this study to treat simulated wastewaters prepared from black liquor from a hardwood Kraft pulp mill in Taiwan. The experimental results showed that the COD and color were primarily removed by direct ozone oxidation and activated carbon adsorption. While the addition of activated carbon could enhance the COD and color removal during ozonation, the addition of hydrogen peroxide improved the color removal only. For the various ozone-based treatment processes, kinetic models were developed to satisfactorily predict the COD and color removal rates. According to the kinetic parameters obtained from the various ozone-based processes, the enhanced COD and color removal of ozonation in the presence of activated carbon was attributed to the regeneration of the activated carbon by ozonation. These kinetic models can be used for reactor design and process design to treat pulping wastewater using ozone-based processes. PMID- 19304381 TI - Sorption of p-nitrophenol by anion-cation modified palygorskite. AB - A series of anion-cation modified palygorskites were prepared by microwave irradiation, and characterized with FT-IR. Compared with natural palygorskite, the p-nitrophenol sorption capacity with anion-cation modified palygorskite was significantly improved. The maximum sorption capacity (137.74 mg g(-1)) was obtained with the mass ratio 1:4 and 3:10 of hexadecyltrimethylammonium bromide to sodium dodecyl sulfate and the surfactant to palygorskite of at 313 K. The sorption equilibrium was in accordance with the Langmuir isothermal sorption equations. Thermodynamic function calculated showed that the sorption was a spontaneous and exothermic process. Free energy of the sorption increased with the increase of the temperature. The sorption of p-nitrophenol followed the pseudo-second-order process (sorption rate constant: 0.00453 mg(-1)min(-1)). PMID- 19304382 TI - Asbestos pollution in an inactive mine: determination of asbestos fibers in the deposit tailings and water. AB - An inactive asbestos mine in Northern Greece, known as MABE, had been operational for 18 years, showing an annual chrysotile production of approximately 100,000 tons. It is estimated that a total of 68 million tons of the mineral serpentine were excavated from the mine, of which 881,000 tons of chrysotile asbestos were produced. The mine deposits are located very near to the river Aliakmonas. The water of the river is extensively used as drinking water, as well as for irrigation. This study estimated the amount of asbestos currently present in the deposits, to at least 1.33 million tons. This is a 10-fold increase since the start of mine operation in 1982. Water samples obtained throughout the river had high chrysotile concentrations, in most cases far exceeding EPA's standard value (7 x 10(6)f/l). Therefore, the mine and the deposits urgently require remediation works, such as removal of large contaminated objects from the mine buildings and re-vegetation of the deposit areas, in order to reduce the asbestos levels in the river water. PMID- 19304383 TI - Selective and efficient tripodal receptors for competitive solvent extraction and bulk liquid membrane transport of Hg2+. AB - A series of tripodal receptors that are different from each other in their end groups were synthesized. The extraction ability and the transport rate of these receptors were evaluated for transition metal ions (Fe(3+), Ni(2+), Cu(2+), Zn(2+) and Hg(2+)). The receptor bearing pyridyl nitrogens extracted Hg(2+) from a buffered aqueous medium containing a mixture of metal ions and transported it into an organic phase with a complete selectivity over the other metal ions. This receptor was effectively used for transporting Hg(2+) from aqueous buffered source phase to aqueous receiving phase through a chloroform membrane. PMID- 19304384 TI - Suppressive effect of magnesium oxide materials on cadmium accumulation in winter wheat grain cultivated in a cadmium-contaminated paddy field under annual rice wheat rotational cultivation. AB - The effectiveness of two kinds of magnesium oxide (MgO) materials, commercial MgO (2250 kg ha(-1)) and a material derived from MgO and magnesium silicate minerals named 'MgO-SH-A' (2250 and 4500 kg ha(-1)1), in suppression of uptake and accumulation of cadmium (Cd) into grain of winter wheat (Triticum aestivum L. cv. Ayahikari) was examined in a Cd-contaminated alluvial paddy field under annual rice-wheat rotational system. The MgO materials were mixed into the plough-layer soil only once prior to the preceding rice cultivation. Cadmium concentration in wheat grain produced from the non-amendment control exceeded the maximum limit of Cd in wheat grain adopted by FAO/WHO (0.2 mg kg(-1)). All of the treatments with the MgO materials significantly lowered plant available Cd fraction in the plough layer soil. However, only the treatment with the commercial MgO at 2250 kg ha(-1) produced wheat grain whose Cd concentration was not only significantly lower than that from the control but also less than 0.2 mg kg(-1). It is suggested that the significant suppressive effect of the commercial MgO on Cd accumulation in wheat grain would be mainly attributed to its high soil neutralizing capacity as compared to that of MgO-SH-A. PMID- 19304385 TI - Effects of some organic pollutants on the exopolysaccharides (EPSs) produced by some Pseudomonas spp. strains. AB - In this study, isolation and characterization of exopolysaccharides produced by Pseudomonas aeruginosa B1, P. fluorescens B5, P. stutzeri B11 and P. putida B15 which had been seen to produce exopolymers of potential interest in biotechnological applications were examined. To initiate the observation of the organic pollutants-polymer interactions, the yield and properties of their extracellular polysaccharide were researched. The exopolysaccharide production by these strains during growth in nutrient broth medium (control) was 41-75 mg L( 1). Also, P. aeruginosa B1, P. fluorescens B5, P. stutzeri B11 and P. putida B15 had exhibited high production of EPSs in presence of various organic pollutants (2,4-D, benzene, BTX and gasoline, respectively) in mineral salt medium (MSM) as a sole carbon source. EPS production by the 4 strains ranged from 40 mg L(-1) to 8 mg L(-1). Monosaccharide composition of EPS produced by these cultures were analyzed by HPLC. Results indicated that EPSs of strains contained neutral sugars and acetylated amino sugars. The neutral sugars in the EPS were mainly composed of glucose, arabinose, glycerol, ribose. The presence of galactronic acid, N acetyl-D-galactosamin and N-acetyl-D-glucosamine indicated the acidic nature of the polysaccharide. Glycerol was the basic structural unit of EPS produced by the strains except P. stutzeri B11 (MSM with 1% BTX). Strain B1 (in NB medium) was found to be composed of neutral sugars (100%) while strain B1 [in MSM medium with 0.2% (v/v) 2.4-D] contained neutral sugars (70.0%), acetylated amino sugars (30.0%). Also, EPS content of strain B5 (in the NB medium) was neutral sugars (99.8%), acetylated amino sugars (0.2%) while the strain B5 [in MSM medium containing the 1% (v/v) benzene] was found to contain neutral sugars (99.9%), acetylated amino sugars (0.1%). However, EPS monomer composition by strain B11 was detected as neutral sugars (99.77%), acetylated amino sugars (0.23%) in NB medium while the strain B11 [in MSM medium with 1% (v/v) BTX] contained neutral sugars (98.2%) and acetylated amino sugars (1.8%). Lastly, in NB medium by strain B15 was found to contain neutral sugars (99.9%) and acetylated amino sugars (0.1%) while in MSM medium in the presence of 1% (v/v) gasoline it was found to contain neutral sugars (83.6%), acetylated amino sugars (16.4%). Monomer composition of control EPSs changed to different structures in the presence of various organic pollutants. Diversities of organic compounds as carbon source affected the monomer composition of EPS produced by some Pseudomonas spp. cultures. PMID- 19304386 TI - Removal of Basic Red 46 dye from aqueous solution by adsorption onto Moroccan clay. AB - In this study, Moroccan crude clay of Safi, which was characterized by X-ray diffraction, is used as adsorbent for the investigation of the adsorption kinetics, isotherms and thermodynamic parameters of the Basic Red 46 (BR46) in aqueous solutions at various dye concentrations, adsorbent masses and pH values. The results showed that the adsorption capacity of the dye increased by initial dye concentration and pH values. Two kinetic models (the pseudo-first-order and the pseudo-second-order) were used to calculate the adsorption rate constants. The adsorption kinetics of the basic dye followed pseudo-second-order model. The experimental data isotherms were analyzed using the Langmuir, Freundlich and Dubinin-Radushkevish equations. The monolayer adsorption capacity for BR46 dye is 54 mg/g of crude clay. Nearly 20 min of contact time was found to be sufficient for the dye adsorption to reach equilibrium. Thermodynamical parameters were also evaluated for the dye-adsorbent system and revealed that the adsorption process is exothermic in nature. PMID- 19304387 TI - Assessment of heavy metal pollution in water using multivariate statistical techniques in an industrial area: a case study from Patancheru, Medak District, Andhra Pradesh, India. AB - Application of different multivariate statistical approaches for the interpretation of data obtained during a monitoring programme of surface and groundwater in Patancheru industrial town near Hyderabad (India) is presented in this study. A number of chemical and pharmaceutical industries have been established since past three decades. Effluents from these industries are reportedly being directly discharged onto surrounding land, irrigation fields and surface water bodies forming point and non-point sources of contamination for groundwater in the study area. Thirteen parameters including trace elements (B, Cr, Mn, Fe, Co, Ni, Zn, As, Sr, Ba and Pb) have been monitored on 53 sampling points from a hydrogeochemical survey conducted in surface and groundwater. Data set thus obtained was treated using R-mode factor analysis (FA) and principal component analysis (PCA). FA identified four factors responsible for data structure explaining 75% of total variance in surface water and two factors in groundwater explaining 85%, and allowed to group selected parameters according to common features. Sr, Ba, Co, Ni and Cr were associated and controlled by mixed origin with similar contribution from anthropogenic and geogenic sources whereas Fe, Mn, As, Pb, Zn, B and Co were derived from anthropogenic activities. This study indicates the necessity and usefulness of multivariate statistical techniques for evaluation and interpretation of the data with a view to get better information about the water quality and design some remedial techniques to prevent the pollution caused by hazardous toxic elements in future. PMID- 19304388 TI - Performance evaluation of low cost adsorbents in reduction of COD in sugar industrial effluent. AB - Studies on reduction of chemical oxygen demand (COD) in effluent from sugar industry have been carried out by employing different absorbents optimizing various parameters, such as initial concentration of adsorbate, pH, adsorbent dosage and contact time. Experimental studies were carried out in batches using metakaolin, tamarind nut carbon and dates nut carbon as adsorbents by keeping initial adsorbent dosage at 1 g l(-1), agitation time over a range of 30-240 min, adsorbent dosage at 100-800 mg l(-1) by varying the pH range from 4 to 10. Characterization of there adsorbents were done using techniques such as Fourier transforms infra red spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscope (SEM). The experimental adsorption data fitted well to Langmuir and Freundlich adsorption isotherms. The isotherms of the adsorbents indicate appreciable adsorption capacity. Higher COD removal was observed at neutral pH conditions. Studies reveal that maximum reduction efficiency of COD takes place using metakaolin as an absorbent at a dosage of 500 mg l(-1) in a contact time of 180 min at pH 7 and it could be used as an efficient absorbent for treating sugar industrial effluent. PMID- 19304389 TI - Pain demands attention from others: the approach/avoidance paradox. PMID- 19304390 TI - Relief of postoperative pain by local anaesthetic infiltration: efficacy for major abdominal and orthopedic surgery. PMID- 19304391 TI - Adverse events in childhood and chronic widespread pain in adult life: Results from the 1958 British Birth Cohort Study. AB - Chronic widespread pain (CWP) is a common and frequently disabling condition. Several studies have shown that early life adversity is associated with CWP in later life; however, the majority are retrospective and suffer from potential recall bias. Using data from the 1958 British Birth Cohort Study, the aim of the current study was to examine, prospectively, the relationship between childhood physical and psychological adversity and CWP in adulthood. At 7 yrs data were collected, by parental report, on physically traumatic events (hospitalisation following a road traffic accident, or for surgery); and factors indicating poor social and psychological environment (periods in local authority care, death of a parent; or parental divorce, alcoholism, or financial hardship). CWP was assessed at 45 yrs using self-completion questionnaires. The relationship between childhood events and CWP was examined using Poisson regression. 7571 individuals provided pain data at 45 yrs (71.5%). There was no association between childhood surgery and CWP in adulthood (relative risk: 1.0; 95%CI: 0.9-1.1). However, children who had been hospitalised following a road traffic accident experienced a significant increase in the risk of future CWP (1.5; 1.05-2.1). Children who had resided in institutional care also experienced an increase in the risk of CWP (1.7; 1.3-2.4) as did those who experienced maternal death (2.0; 1.08-3.7) and familial financial hardship (1.6; 1.3-1.9). Further these associations were not explained by adult psychological distress or social class. To prevent long-term consequences of adverse childhood events, future research should study the mechanisms, in particular the biological mechanisms, underlying these relationships. PMID- 19304392 TI - Psychological determinants of problematic outcomes following Total Knee Arthroplasty. AB - The primary objective of the present study was to examine the role of pain related psychological factors in predicting pain and disability following Total Knee Arthroplasty (TKA). The study sample consisted of 75 (46 women, 29 men) individuals with osteoarthritis of the knee who were scheduled for TKA. Measures of pain severity, pain catastrophizing, depression, and pain-related fears of movement were completed prior to surgery. Participants completed measures of pain severity and self-reported disability 6 weeks following surgery. Consistent with previous research, cross-sectional analyses revealed significant correlations among measures of pre-surgical pain severity, pain catastrophizing, depression and pain-related fears of movement. Prospective analyses revealed that pre surgical pain severity and pain catastrophizing were unique predictors of post surgical pain severity (6-week follow-up). Pain-related fears of movement were predictors of post-surgical functional difficulties in univariate analyses, but not when controlling for pre-surgical co-morbidities (e.g. back pain). The results of this study add to a growing literature highlighting the prognostic value of psychological variables in the prediction of post-surgical health outcomes. The results support the view that the psychological determinants of post-surgical pain severity differ from the psychological determinants of post surgical disability. The results suggest that interventions designed to specifically target pain-related psychological risk factors might improve post surgical outcomes. PMID- 19304393 TI - Two novel SCN9A mutations causing insensitivity to pain. AB - The sensation of pain is important and there may be serious consequences if it is missing. Recently, the genetic basis for a channelopathy characterised by a congenital inability to experience pain has been described and channelopathy associated insensitivity to pain has been proposed as a suitable name for this condition. Different mutations in the SCN9A gene causing loss of function of the voltage-gated sodium channel Nav1.7 have been reported in patients with this rare disease. Here we describe a woman with insensitivity to pain with two novel mutations in the SCN9A gene, coding for the Nav1.7 channel. We also discuss the finding of anosmia which apparently is a common feature in these patients. PMID- 19304394 TI - Behaviour of TEM metal grids during in-situ heating experiments. AB - The stability of Ni, Cu, Mo and Au transmission electron microscope (TEM) grids coated with ultra-thin amorphous carbon (alpha-C) or silicon monoxide film is examined by in-situ heating up to a temperature in the range 500-850 degrees C in a transmission electron microscope. It is demonstrated that some grids can generate nano-particles either due to the surface diffusion of metal atoms on amorphous film or due to the metal evaporation/redeposition. The emergence of nano-particles can complicate experimental observations, particularly in in-situ heating studies of dynamic behaviours of nano-materials in TEM. The most widely used Cu grid covered with amorphous carbon is unstable, and numerous Cu nano particles start to form once the heating temperature reaches 600 degrees C. In the case of Ni grid covered with alpha-C film, a large number of Ni nano-crystals occur immediately when the temperature approaches 600 degrees C, accompanied by the graphitization of amorphous carbon. In contrast, both Mo and Au grids covered with alpha-C film exhibit good stability at elevated temperature, for instance, up to 680 and 850 degrees C for Mo and Au, respectively, and any other metal nano particles are detected. Cu grid covered Si monoxide thin film is stable up to 550 degrees C, but Si nano-crystals appear under intensive electron beam. The generated nano-particles are well characterized by spectroscopic techniques (EDXS/EELS) and high-resolution TEM. The mechanism of nano-particle formation is addressed based on the interactions between the metal grid and the amorphous carbon film and on the sublimation of metal. PMID- 19304395 TI - Existence of Angiostrongylus cantonensis in rats (Rattus norvegicus) in Grenada, West Indies. AB - The zoonotic rat lung worm, Angiostrongylus cantonensis is endemic to Asia, North America, Africa and Australia. The parasite is expanding geographically and has recently been recorded in some of the Greater Antilles in the northern part of the Caribbean. In this study A. cantonensis is reported for the first time in the Lesser Antilles in one of the southernmost islands, Grenada. Between September 2005 and September 2006, 192 rats (Rattus norvegicus) were trapped throughout the island. The rats were anesthetized, exsanguinated, necropsied and the lungs were fixed whole in 10% buffered formalin, trimmed, processed, cut at 3microm, stained with hematoxylin and eosin and examined microscopically. A total of 45 (23.4%) of the 192 rats examined were found to be infected with A. cantonensis and adult worms were found in the cardiopulmonary system of one of the rats. Microscopically, pulmonic lesions, consisting of pulmonary thrombosis, hypertrophy of pulmonary arteries and granulomatous pneumonia were associated with intralesional adults, larvae and embryonated eggs of A. cantonensis. An incidental finding of variably sized (2-7mm) solitary to multiple cysts containing larvae of Taenia taeniaformis were seen in the livers of 57 rats. This report of A. cantonensis in Grenada provides evidence of the further global expansion of this important zoonotic parasite and the public health implications of this discovery is discussed. PMID- 19304396 TI - Autochthonous Angiostrongylus vasorum infection in dogs in The Netherlands. AB - Until recently, autochthonous infections with Angiostrongylus vasorum were not thought to occur in The Netherlands. However, in 2007 four dogs were diagnosed with angiostrongylosis. Three of them had never left the country. Subsequently, faecal samples from 485 dogs living in the same region where the positive dogs had been diagnosed in 2007 were collected through veterinarians, hunting associations and kennels between January and April 2008. A. vasorum larvae were found in faecal samples of four dogs. Three of those had never been outside the country. Though only few samples were found positive, A. vasorum is now considered autochthonous in The Netherlands. PMID- 19304397 TI - The direct hospitalisation costs of paediatric scalds: 2-year results of a prospective case series. AB - OBJECTIVE: To reveal the characteristic and distribution of length of hospital stay (LOS) and direct hospitalisation costs of paediatric scald. METHODS: A prospective case series observation was performed from January 2005 to December 2006 at the Burn Center, Changhai Hospital, Shanghai, China. The information, such as demographics, clinical diagnosis and treatments since admission, of the paediatric scald patients included in the series was recorded. The direct cost of a treatment event was recorded into the price system when it was incurred. All cost data were summarised on completion of the study. The distribution of LOS and the hospitalisation costs were recorded by gender, age, total burn area, depth of burn, blood transfusion and patterns of treatment. Mann-Whitney signed-rank test was used to assess the differences between continuous, non-normally distributed variables, and multiple linear regression was used to model LOS and direct hospitalisation costs. Statistical analyses were undertaken with SPSS 15.0 statistical software. RESULTS: Patients aged 3 years or less accounted for more than half of the total LOS and hospitalisation costs, patients with burn area less than 10%TBSA (total burn surface area) accounted for more than 70% of the total LOS and more than half of the hospitalisation costs and patients with second-degree burn accounted for more than 78% of the total LOS and hospitalisation costs. Depth of burn, area of burn, patterns of treatment and blood transfusion were independent predictors of LOS; whereas LOS, area of burn and blood transfusion were independent predictors of hospitalisation costs. CONCLUSION: Paediatric scalds have particular characteristics in terms of distribution of LOS and direct hospitalisation costs and the factors influencing them. The data presented in this study should assist burn care practitioners and hospital epidemiologists estimate and compare the economic burden of paediatric burns at other institutions; it may also be useful in resource allocation and cost-effectiveness analysis of treatment versus prevention strategies. PMID- 19304398 TI - Burned by a battery-coin short circuit: old concept for a new burn. PMID- 19304399 TI - Iatrogenic burns. PMID- 19304400 TI - Absence of a normal cortisol awakening response (CAR) in adolescent males with Asperger syndrome (AS). AB - In addition to abnormalities in social and communication development, a 'need for sameness' and 'resistance to change' are features of autistic spectrum disorders first identified by Kanner in 1943. Our ability to react to change is modulated by the hypothalamic-pituitary-adrenal (HPA) axis, a feature of which is a dramatic increase in cortisol upon waking, the Cortisol Awakening Response (CAR). This study examined whether the CAR was evident in 20 adolescent males with Asperger Syndrome (AS) and 18 age-matched typically developing (TD) controls (aged 11-16). Whilst a significant CAR was evidenced in the TD control group, this was not the case for those with AS. A normal diurnal decrease in cortisol, however, was evident in both groups. The implication that individuals with AS may have an impaired response to change in their environment due to a refractory HPA axis is discussed. PMID- 19304401 TI - Proton pump inhibitors as a treatment method for type II diabetes. AB - Recent reports have hypothesized a role for exogenously administered gastrin in regulating beta cell function or activity. We surmised that a class of agents, proton pump inhibitors (omeprazole, esomeprazole, pantoprazole, rabeprazole, lansoprazole), known to increase serum gastrin levels could have such an effect, and that data might be available in our database which could address such an effect. We examined our electronic database to obtain glycohemoglobin (HgbA1c) levels measured in the past two years from all type II diabetics and extracted from those diabetics who were concurrently taking a proton pump inhibitor. A comparison of these groups showed an average HgbA1c of 7.6% for type II diabetics not taking a proton pump inhibitor (n=282) and an average HgbA1c of 7.0% for type II diabetics concurrently taking a proton pump inhibitor (n=65), T=-3.61, p=0.002. These data support the hypothesis that proton pump inhibitors can be used to treat type II diabetes. PMID- 19304402 TI - Customized computed tomography-based boost volumes in breast-conserving therapy: use of three-dimensional histologic information for clinical target volume margins. AB - PURPOSE: To determine the difference in size between computed tomography (CT) based irradiated boost volumes and simulator-based irradiated volumes in patients treated with breast-conserving therapy and to analyze whether the use of anisotropic three-dimensional clinical target volume (CTV) margins using the histologically determined free resection margins allows for a significant reduction of the CT-based boost volumes. PATIENTS AND METHODS: The CT data from 49 patients were used to delineate a planning target volume (PTV) with isotropic CTV margins and to delineate a PTV(sim) that mimicked the PTV as delineated in the era of conventional simulation. For 17 patients, a PTV with anisotropic CTV margins was defined by applying customized three-dimensional CTV margins, according to the free excision margins in six directions. Boost treatment plans consisted of conformal portals for the CT-based PTVs and rectangular fields for the PTV(sim). RESULTS: The irradiated volume (volume receiving > or =95% of the prescribed dose [V(95)]) for the PTV with isotropic CTV margins was 1.6 times greater than that for the PTV(sim): 228 cm(3) vs. 147 cm(3) (p < .001). For the 17 patients with a PTV with anisotropic CTV margins, the V(95) was similar to the V(95) for the PTV(sim) (190 cm(3) vs. 162 cm(3); p = NS). The main determinant for the irradiated volume was the size of the excision cavity (p < .001), which was mainly related to the interval between surgery and the planning CT scan (p = .029). CONCLUSION: CT-based PTVs with isotropic margins for the CTV yield much greater irradiated volumes than fluoroscopically based PTVs. Applying individualized anisotropic CTV margins allowed for a significant reduction of the irradiated boost volume. PMID- 19304403 TI - Molecular genetic changes associated with colorectal carcinogenesis are not prognostic for tumor regression following preoperative chemoradiation of rectal carcinoma. AB - PURPOSE: Preoperative chemotherapy and radiation has become the standard of care for many patients with rectal cancer. The therapy may have toxicity and delays definitive surgery. It would therefore be desirable to identify those cancers that will not regress with preoperative therapy. We assessed a series of rectal cancers for the molecular changes of loss of heterozygosity of the APC and DCC genes, K-ras mutations, and microsatellite instability, changes that have clearly been associated with rectal carcinogenesis. METHODS AND MATERIALS: Diagnostic colonoscopic biopsies from 53 patients who received preoperative chemotherapy and radiation were assayed using polymerase chain reaction techniques followed by single-stranded conformation polymorphism and DNA sequencing. Regression of the primary tumor was evaluated using the surgically removed specimen. RESULTS: Twenty-three lesions (45%) were found to have a high degree of regression. None of the molecular changes were useful as indicators of regression. CONCLUSIONS: Recognized molecular changes critical for rectal carcinogenesis including APC and DCC loss of heterozygosity, K-ras mutations, and microsatellite instability are not useful as indicators of tumor regression following chemoradiation for rectal carcinoma. PMID- 19304404 TI - Effect of interval to definitive breast surgery on clinical presentation and survival in early-stage invasive breast cancer. AB - PURPOSE: To examine the effect of clinical presentation and interval to breast surgery on local recurrence and survival in early-stage breast cancer. METHODS AND MATERIALS: The data from 397 patients with Stage T1-T2N0 breast carcinoma treated with conservative surgery and breast radiotherapy between 1985 and 1992 were reviewed at the London Regional Cancer Program. The clinical presentation consisted of a mammogram finding or a palpable lump. The intervals from clinical presentation to definitive breast surgery used for analysis were 0-4, >4-12, and >12 weeks. The Kaplan-Meier estimates of the time to local recurrence, disease free survival, and cause-specific survival were determined for the three groups. Cox regression analysis was used to evaluate the effect of clinical presentation and interval to definitive surgery on survival. RESULTS: The median follow-up was 11.2 years. No statistically significant difference was found in local recurrence as a function of the interval to definitive surgery (p = .424). A significant difference was noted in disease-free survival (p = .040) and cause-specific survival (p = .006) with an interval of >12 weeks to definitive breast surgery. However, the interval to definitive surgery was dependent on the presentation for cause-specific survival, with a substantial effect for patients with a mammographic presentation and a negligible effect for patients with a lump presentation (interaction p = .041). CONCLUSION: The results of this study suggest that an interval of >12 weeks to breast surgery might be associated with decreased survival for patients with a mammographic presentation, but it appeared to have no effect on survival for patients presenting with a palpable breast lump. PMID- 19304405 TI - Individual positioning: a comparative study of adjuvant breast radiotherapy in the prone versus supine position. AB - PURPOSE: To study breast radiotherapy in the prone vs. supine positions through dosimetry and clinical implementation. METHODS AND MATERIALS: Conformal radiotherapy plans in 61 patients requiring only breast irradiation were developed for both the prone and supine positions. After evaluation of the of the first 20 plan pairs, the patients were irradiated in the prone or supine position in a randomized fashion. These cases were analyzed for repositioning accuracy and skin reactions related to treatment position and patient characteristics. RESULTS: The planning target volume covered with 47.5-53.5 Gy in the prone vs. the supine position was 85.1% +/- 4.2% vs. 89.2 +/- 2.2%, respectively (p < 0.0001). Radiation exposure of the ipsilateral lung, expressed in terms of the mean lung dose and the V(20Gy), was dramatically lower in the prone vs. supine position (p < 0.0001), but the doses to the heart did not differ. There was no difference in the need to correct positioning during radiotherapy, but the extent of displacement was significantly higher in the prone vs. supine position (p = 0.021). The repositioning accuracy in the prone position exhibited an improvement over time and did not depend on any patient-related parameters. Significantly more radiodermatitis of Grade 1-2 developed following prone vs. supine irradiation (p = 0.025). CONCLUSIONS: Conformal breast radiotherapy is feasible in the prone position. Its primary advantage is the substantially lower radiation dose to the ipsilateral lung. The higher dose inhomogeneity and increased rate of Grade 1-2 skin toxicity, however, may be of concern. PMID- 19304406 TI - Mucositis versus tumor control: the therapeutic index of adding chemotherapy to irradiation of head and neck cancer. AB - PURPOSE: To determine whether the addition of concurrent chemotherapy to radiation for head and neck cancer (HNSCC) improves the therapeutic ratio regarding tumor control vs. mucositis. METHODS AND MATERIALS: Data were taken from 14 randomized trials of radiation with or without concurrent chemotherapy for HNSCC. Mucositis-bioequivalent dose (mBED) was computed for each study using mBED = D [1 + d/(alpha/beta)] - 0.693(T - Tk)/Tp. An "S-value," relating the increase in the rate of Grade 3 (confluent) mucositis to the increase in mBED with radiation alone, was determined using data from trials of radiation alone with altered fractionation. We then determined the difference in the rate of mucositis and used the S-value to estimate the apparent difference in mBED in the chemoradiation and radiation alone arms for each trial. After accounting for differences in the radiation schedules, we estimated the mBED attributable to adding chemotherapy and compared it with previously published estimates of increases in tumor BED. RESULTS: Computed S-values ranged from 0.4 to 1.7. For S = 1, the mean increase in mBED attributable to chemotherapy was 8.3 Gy(10) (SD = 6.4). The average difference between tumor-BED and mBED was 2.8 Gy(10) (SD = 6.0). Increasing the S-value decreases the estimated increase in mBED due to chemotherapy. CONCLUSIONS: Concurrent chemotherapy improves the therapeutic index for radiation of HNSCC. Further refinements are needed in quantifying the therapeutic gain attributable to specific radiosensitizing agents in clinical trials, notably better and more consistent reporting of treatment sequelae. PMID- 19304407 TI - Neoadjuvant treatment does not influence perioperative outcome in rectal cancer surgery. AB - PURPOSE: To identify the risk factors for perioperative morbidity in patients undergoing resection of primary rectal cancer, with a specific focus on the effect of neoadjuvant therapy. METHODS AND MATERIALS: This exploratory analysis of prospectively collected data included all patients who underwent anterior resection/low anterior resection or abdominoperineal resection for primary rectal cancer between October 2001 and October 2006. The study endpoints were perioperative surgical and medical morbidity. Univariate and multivariate analyses of potential risk factors were performed. RESULTS: A total of 485 patients were included in this study; 425 patients (88%) underwent a sphincter saving anterior resection/low anterior resection, 47 (10%) abdominoperineal resection, and 13 (2%) multivisceral resection. Neoadjuvant chemoradiotherapy was performed in 100 patients (21%), and 168 (35%) underwent neoadjuvant short-term radiotherapy (5 x 5 Gy). Patient age and operative time were independently associated with perioperative morbidity, and operative time, body mass index >27 kg/m(2) (overweight), and resection type were associated with surgical morbidity. Age and a history of smoking were confirmed as independent prognostic risk factors for medical complications. Neoadjuvant therapy was not associated with a worse outcome. CONCLUSION: The results of this prospective study have identified several risk factors associated with an adverse perioperative outcome after rectal cancer surgery. In addition, neoadjuvant therapy was not associated with increased perioperative complications. PMID- 19304408 TI - A prospective study of hypofractionated proton beam therapy for patients with hepatocellular carcinoma. AB - PURPOSE: To evaluate the efficacy and safety of hypofractionated proton beam therapy for patients with hepatocellular carcinoma (HCC). METHODS AND MATERIALS: Between September 2001 and August 2004, 51 patients with HCC greater than 2 cm away from the porta hepatis or gastrointestinal tract were treated with proton beam therapy to 66 Gy-equivalents (GyE) in 10 fractions. RESULTS: Overall survival rates were 49.2 and 38.7% at 3 and 5 years after treatment. Local control rates were 94.5 and 87.8% at 3 and 5 years after treatment. Posttreatment serum alpha-fetoprotein values were significantly reduced when compared with pretreatment values (p < 0.0001). Patients experienced only minor acute reactions of Grade 1 or less, and 3 patients experienced late sequelae of Grade 2 or higher. However, there were no treatment-related deaths. CONCLUSIONS: Hypofractionated proton beam therapy is safe and well-tolerated by patients with HCC located greater than 2 cm away from the porta hepatis or gastrointestinal tract and may be effective alternative to other modalities. PMID- 19304410 TI - [Symptoms of HELLP syndrome due to vitamin B12 deficiency: report of seven cases]. AB - INTRODUCTION: HELLP syndrome is characterized by generalised thrombotic microangiopathy predominant in the liver. Many investigators consider HELLP syndrome to be a variant of severe preeclampsia. Several other conditions have similar laboratory findings in common with HELLP syndrome and should be eliminated before pregnancy termination. PATIENTS AND METHODS: The authors report seven observations of patients with biological criteria that mimic HELLP syndrome secondary to severe vitamin B12 deficiency. CONCLUSION: Vitamin B12 or B9 deficiency developed during pregnancy have laboratory findings similar to those of HELLP syndrome. PMID- 19304409 TI - Salvage treatment improved survival of patients with relapsed extranodal natural killer/t-cell lymphoma, nasal type. AB - PURPOSE: To evaluate the clinical outcome of salvage treatment for patients with relapsed natural killer (NK)/T-cell lymphoma, nasal type. METHODS AND MATERIALS: Forty-four patients who had achieved complete response during initial treatment and experienced histologically proven relapse were reviewed. Twenty-nine of them received salvage treatment with radiotherapy (RT) alone (n = 7), chemotherapy (CT) alone (n = 10), or both RT and CT (n = 12); the other 15 patients received best supportive care alone. RESULTS: The estimated 5-year overall survival (OS) rate for patients with or without salvage treatment was 37.8% vs. 0 (p < 0.0001), respectively. Salvage CT did not improve survival of relapsed Stage IE and IIE patients. Among relapsed Stage IIIE and IVE patients who received salvage treatment, RT developed significantly better survival when compared with that of non-RT (1-year OS, 62.5% vs. 0, p = 0.006). Relapsed Ann Arbor stage and receiving salvage treatment were found to be significant factors influencing OS at both univariate and multivariate levels. CONCLUSIONS: Salvage treatment improved survival in patients with relapsed NK/T-cell lymphoma, nasal type. Salvage RT may play an important role in salvage treatment of relapsed extranodal NK/T-cell lymphoma. PMID- 19304411 TI - [Ovarian metastasis of breast cancer: a case report. Role of cytoreductive surgery]. AB - The ovaries are a common metastatic site for breast cancer. The diagnosis and treatment of ovarian masses from a metastatic breast cancer are difficult. The complete resection of these metastatic masses seems to give a benefit in terms of global survival. This benefit depends on the residual tumoral volume and on the free interval between initial breast cancer diagnosis and apparition of the metastatic ovarian masses. We discuss the treatment of a patient with ovarian metastasis as first sign of a metastatic breast cancer. PMID- 19304412 TI - The 1.8kb DNA fragment formerly confirmed as Mycoplasma suis (M. suis) specific was originated from the porcine genome. PMID- 19304413 TI - Endocrine changes during pregnancy, parturition and post-partum in guanacos (Lama guanicoe). AB - Plasma concentrations of progesterone (P4), estradiol-17beta (E2), estrone (E1) and estrone sulfate (E1S) were measured during gestation in eight guanacos kept in captivity. Gestational length was 346.1+/-9.8 days. P4 plasma concentrations increased after ovulation and remained elevated until parturition. However, during the last 4 weeks of gestation, a gradual decrease from 4.17x1.17(+/ 1)nmol/L to 2.02x1.95(+/-1)nmol/L on day 5 before parturition was observed, followed by a more abrupt final decline to baseline concentrations which were reached on the day after parturition. Mean E2 plasma concentrations started to increase during the eighth month of gestation, and were significantly elevated up to maximum concentrations of 484.7x1.21(+/-1)pmol/L during the last 2 months of pregnancy. Concentrations returned to baseline during the last 2 days of gestation. An increase of E1S concentrations (p<0.01) was observed in the eleventh month of gestation. Mean E1S concentrations remained rather constant during the last 3 weeks of gestation between 4 to 8nmol/L until parturition, when a steep precipitous decline was observed. E1 concentrations were slightly elevated during the last 4 weeks of gestation, however, maximum concentrations did not exceed 1.5nmol/L. The results show distinct species specific features of gestational steroid hormone profiles in the guanaco in comparison to domestic South American camelids, such as a more pronounced gradual prepartal decrease of P4 concentrations prior to the final decline to baseline, and clearly lesser E1S concentrations during the last 4 weeks of gestation, which lack a continuous prepartal increase. PMID- 19304414 TI - The effect of trehalose on post-thaw viability and fertility of European brown hare (Lepus europaeus Pallas, 1778) spermatozoa. AB - The aim of the study was to assess the effect of a different trehalose concentration on the post-thaw viability and fertility of European brown hare spermatozoa. The semen was collected under general anaesthesia with electroejaculation method from 4 males. Immediately after collection, the semen was diluted with an extender of the following composition: Tris 250mM, citric acid 80mM, glucose 70mM, DMSO 1.0M, egg yolk (17%, v/v), and kanamycin (80mg/l) Protocol I. In Protocols II and III, respectively, 50mM and 100mM of trehalose were added to the extender. Immediately after thawing and after 3h incubation at 37 degrees C, motility characteristics of frozen/thawed semen were assessed with computer-assisted semen analysis system, and a percentage of viable, acrosome intact spermatozoa was evaluated using flow cytometry. Immediately after thawing spermatozoa motility (MOT), average path velocity (VAP), straight velocity (VSL) and curvilinear velocity (VCL) were the highest in the semen frozen without addition of trehalose (P<0.01). After 3h of incubation, MOT and spermatozoa with progressive motility (PMOT) were the lowest in the semen frozen with supplementation of 100mM of trehalose (P<0.01) and VAP, VSL, VCL and amplitude of lateral head displacement (ALH) were significantly lower in the semen frozen with supplementation of 50mM and 100mM of trehalose compared to the semen frozen without the addition of trehalose (P<0.01), which indicates an unfavourable effect of trehalose on the motility characteristics of European brown hare spermatozoa. However, the effect of trehalose on a percentage of viable, acrosome intact spermatozoa was not observed. As a result of artificial insemination, 54.55% females became pregnant after insemination with the semen frozen according to Protocol I, 72.73% and 50% females became pregnant after insemination with the semen supplemented with 50mM and 100mM of trehalose, respectively. The number of young born was 1.67+/-0.52, 1.75+/-1.04 and 1.60+/-0.55, in each group, respectively. There were not any significant differences in the results of artificial insemination between groups. Summing up, it should be stated that the addition of trehalose to the extender did not have a favourable effect on post thaw viability of European brown hare spermatozoa and an influence of trehalose on the results of artificial insemination was not found, either. PMID- 19304415 TI - Proliferation and apoptosis of male germ cells in captive Atlantic bluefin tuna (Thunnus thynnus L.) treated with gonadotropin-releasing hormone agonist (GnRHa). AB - The effects of administration of gonadotropin-releasing hormone agonist (GnRHa) on proliferation and apoptosis of male germ cells were evaluated on Atlantic bluefin tuna (Thunnus thynnus L.) reared in captivity. Fish (n=19) were treated with a sustained-release delivery system loaded with GnRHa during the natural spawning season of 2004 and 2005 (June-July). Untreated Control fish (n=17) and adult wild spawners were used for comparison. Fish were sacrificed 2-8 d after GnRHa implantation and body weight and gonad weight were recorded, and gonads and blood were taken. Germ cell proliferation and apoptosis were evaluated through the immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and the terminal deoxynucleotidyl transferase-mediated d'UTP nick end labelling (TUNEL) method, respectively. Plasma 11 ketotestosterone (11-KT) levels were measured using an ELISA method. Mean gonado-somatic index and seminiferous lobule diameter did not differ between GnRHa-treated and Control fish, and were significantly lower in captive-reared individuals than in wild spawners. Significant increases in 11-KT plasma levels and spermatogonial mitosis, along with a reduction of germ cell apoptosis were demonstrated in GnRHa-treated fish compared to Controls. The results suggest that GnRHa administration was effective in enhancing germ cell proliferation and reducing apoptosis in captive males through the stimulation of luteinizing hormone (LH) release and testicular 11-KT production. PMID- 19304416 TI - Nutritional status influences reproductive seasonality in Creole goats: 1. Ovarian activity during seasonal reproductive transitions. AB - The objective was to determine the effect of body energy stores, evaluated by a body mass index (BMI), and food intake (FI), on the length of the anovulatory period and ovarian activity during the seasonal reproductive transitions in Creole goats. Non-pregnant, non-lactating Creole goats (n=28) were fed to induce two different BMI conditions: Greater (GBMI; n=15), and Lesser (LBMI; n=13). Each BMI group was divided into two sub-groups, which were either feed restricted (FR) or non-feed restricted (NFR). Goats in the NFR groups received a diet containing 100% of the daily maintenance requirements (basal diet), while restricted goats were subjected to alternated periods, receiving 100% (11d) and 60% (10d) of the basal diet, during the entire experimental period. The experiment started after does were treated to synchronize time of estrus. Serum progesterone was determined in samples obtained twice a week, and used as a criterion for determining ovulations. During the transition to the anovulatory period three transrectal ovarian ultrasonographic scans were performed in a sub-group of 12 goats (n=3 for each treatment combination). The diameter of the largest follicle (LFD) and the total number of antral follicles >or=2mm (TAF) were recorded. Ultrasonographic ovarian scans were performed at 21, 42 and 63 days after the beginning of the experiment, concurrently with the end of each feed restriction period. The variables of response associated with ovulation were not influenced by BMI or BMIxFI interaction. However, FI influenced length of anovulatory season, as the anovulatory period was 30d longer (P<0.05) in the FR group as compared with the NFR group. Independently of treatments, TAF and LFD decreased from the first to the third ultrasonographic ovarian scan (13.2, 10.8 and 4.4 follicles; 3.7, 2.7 and 2.3mm). Nevertheless, in PER 1 the number of TAF was greater (P<0.05) in the FR as compared with NFR group and the GBMI group had a larger LFD (P<0.05) as compared to the LBMI group. It is concluded, that temporal restriction in feed intake could affect the time of cessation and initiation of ovulations during the periods of transition to seasonal anestrus and return to estrous activity, and increase the length of the anovulatory period. In addition, ovarian follicular development during transition into the anovulatory period is differentially influenced by food intake and the status of body energy stores. PMID- 19304417 TI - Characterization of seasonal reproduction patterns in female pichis Zaedyus pichiy (Xenarthra: Dasypodidae) estimated by fecal sex steroid metabolites and ovarian histology. AB - Reproductive strategies vary considerably among species, but most studies have focused on a very limited number of mammalian species. Knowledge of the reproductive cycle and behavior is essential for developing and implementing in situ and ex situ conservation strategies for threatened and endangered species. This study aimed at characterizing the seasonal reproductive pattern of female pichis Zaedyus pichiy, a threatened small armadillo native to arid regions of Argentina and Chile, through direct observations, histological studies, and by measuring fecal immunoreactive estrogens, progestagens and glucocorticoids in 10 wild-born, captive pichis and in free-ranging individuals. Results suggest that pichis are seasonal breeders that give birth to one yearly litter of 1-2 offspring, which do not leave the burrow until they are weaned at approximately 37 days. Ovarian follicular growth seems to occur throughout the year. Fecal progestagen, estrogen and glucocorticoid concentrations were minimal during the first half of pregnancy, increased to peak concentrations of up to 3500, 200 and 200ng/g dry feces, respectively, and decreased before parturition. Postpartum progestagen concentrations were greater in lactating females than females that aborted or did not raise their offspring (p<0.0001), which is probably related to an elevated corticosteroid synthesis that contributes to maintain lactation, given that fecal glucocorticoid concentrations were of similar pattern. Observations of a second pregnancy after late abortion or death of the newborn litter and sustained follicular growth during pregnancy and lactation suggest that female pichis can become receptive briefly after having lost their litter. Fecal estrogen and progestagen concentrations of non-pregnant, non-lactating females did not have a well-defined hormonal cyclic pattern, and corpora lutea were only observed in pregnant females. PMID- 19304418 TI - Heroin-associated myocardial damages--conventional and immunohistochemical investigations. AB - Well-known complications related to drug abuse are myocardial insufficiency, myocardial infarction, endocarditis, myocarditis, aortic dissection, neurologic damages, ischemic colitis, thrombotic phenomenons, renal infarction and acute liver failure. Furthermore, microfocal fibrosis of the myocardium is found in stimulant abuse. The origin of myocardial fibrosis associated with opiate abuse (endocarditis, myocarditis, embolism) is still unclear. This question shall be investigated using immunohistochemical staining for early diagnosis of myocarditis. A quantification of myocardial interstitial leucocytic infiltrates was accomplished in 21 chronic drug abusers who died of heroin/morphine intoxication and compared to 15 normal subjects who died suddenly due to non cardiac causes of death without intoxication (e.g. traffic accidents, head trauma). Toxicological investigations were performed and in addition, blood samples were checked to clarify the status of HIV, hepatitis A, B and C in both groups. To verify signs of inflammation, myocardial specimen from different locations were investigated with conventional histological stainings and immunohistochemical techniques for characterization and quantification of interstitial myocardial leucocytes, T-lymphocytes and macrophages. The number of cells were found up to fivefold increased in heroin addicts compared to the control group without reaching the cut-off values for immunohistochemically based diagnosis of myocarditis. PMID- 19304419 TI - Velopharyngeal stenosis, a late complication of radiotherapy. AB - Choanal stenosis has recently been recognized as a late complication of radiation therapy for nasopharyngeal carcinoma. The management of velopharyngeal stenosis is challenging with high risk of restenosis. We report a case of velopharyngeal stenosis post-radiotherapy and illustrated the use of mitomycin-C to prevent restenosis. Mitomycin-C application has being shown useful adjunct to surgical technique in managing nasopharyngeal stenosis for surgeons. PMID- 19304420 TI - Efficacy and tolerability of adjunctive therapy with zonisamide in childhood intractable epilepsy. AB - PURPOSE: This study investigated the efficacy and safety of zonisamide (ZNS) adjunctive therapy in children with intractable epilepsy to existing antiepileptic drugs (AEDs). METHODS: A clinical retrospective study was performed from 2003 to 2005 at two tertiary epilepsy centers. We reviewed the data from 163 children (107 boys and 56 girls) who experienced more than four seizures per month, whose seizures were intractable to an initial 2 or more AEDs, and could be followed up for at least 6months after ZNS adjunctive therapy initiation. Efficacy was estimated by seizure reduction rate according to seizure types including infantile spasms, and adverse events were also measured. RESULTS: Seventy-nine patients (48.5%) out of 163 patients experienced a reduction in seizure frequency of more than 50%, and 25 patients (15.3%) became seizure-free. The rate of seizure reduction greater than 50% in children with partial seizures was 40.5% (17/42) and in children with generalized seizures was 51.2% (62/121). Of 36 patients who manifested mainly myoclonic seizures, 20 patients (55.6%) showed a seizure reduction of more than 50% and 9 patients (25.0%) were seizure free. Mean maintenance dosage of drug was 8.2mg/kg/day (range 5.0-16.0mg/kg/day). Adverse events were documented in 15 children (9.2%), including somnolence (8 patients), fatigue, and anorexia, but all were transient and successfully managed. One patient discontinued ZNS therapy due to acute pancreatitis. CONCLUSION: ZNS adjunctive therapy is an effective and safe treatment in various childhood intractable epilepsy. PMID- 19304421 TI - The expanding phenotype of GLUT1-deficiency syndrome. AB - Transport of glucose from the bloodstream across the blood-brain barrier to the central nervous system is facilitated by glucose transport protein type 1 (GLUT1), the first member of the solute carrier family 2 (SLC2). Heterozygous mutations in the GLUT1/SLC2A1 gene, occurring de novo or inherited as an autosomal dominant trait, result in cerebral energy failure and a clinical condition termed GLUT1-deficiency syndrome (GLUT1-DS). Clinical features usually comprise motor and mental developmental delay, seizures with infantile onset, deceleration of head growth often resulting in acquired microcephaly, and a movement disorder with ataxia, dystonia, and spasticity. Subsequent to the delineation of this classic phenotype the variability of signs and symptoms in GLUT1-DS is being recognized. Patients with (i) carbohydrate-responsive symptoms, with (ii) predominant ataxia or dystonia, but without seizures, and with (iii) paroxysmal exertion-induced dyskinesia and seizures have been reported. Common laboratory hallmark in all phenotypes is the reduced glucose level in cerebrospinal fluid with lowered CSF-to-blood glucose ratio. Treatment with a ketogenic diet results in marked improvement of seizures and movement disorders. PMID- 19304422 TI - [Contraceptive use and incidence of pregnancy among women after HIV testing in Abidjan, Ivory Coast]. AB - BACKGROUND: Within the framework of programs for the prevention of mother-to child HIV transmission, women who discover their HIV-infection during their pregnancy receive perinatal interventions in order to reduce the risk of HIV transmission to the child. They also receive family planning counselling and free contraceptives in order to avoid a new pregnancy. In this study, we compared contraceptive use and pregnancy incidence between HIV-positive and HIV-negative women who were offered HIV counselling and testing during a program of prevention of mother-to-child HIV transmission. METHODS: In the Ditrame Plus program in Abidjan, 546 HIV-positive and 393 HIV-negative women were HIV-tested prenatally and followed up 2 years after delivery. At each post-partum visit, proportions of contraceptive use were noted, by method. The pregnancy incidence was calculated as the number of pregnancies for 100 women-years at risk. Factors related to the arrival of a new pregnancy were analyzed by Cox model. RESULTS: Between 6 and 24 months post-partum, proportions of women using modern contraception varied from 52 to 65% among HIV-positive women, and from 65 to 75% among HIV-negative women. Pregnancy incidence for 100 women-years at risk was 5.70 (95%CI: 4.17-7.23) and 4.37 (95%CI : 2.83-5.91) (p=0.237) and unwanted pregnancy incidence was 1.07 (95%CI: 0.41-1.73) and 2.39 (95%CI: 1.25-3.53) (p=0.023), respectively among HIV positive and HIV-negative women. The end of post-partum abstinence, the death of the index child and the end of breastfeeding were positively linked to the arrival of a new pregnancy in the post-partum period. CONCLUSION: Among these women prenatally HIV-tested, family planning counselling and regular follow-up was accompanied by a high rate of contraceptive use after delivery, and consecutively to a low pregnancy incidence irrespective of serostatus. In particular, HIV-positive women had fewer unwanted pregnancies than HIV-negative women. Integration of adequate family planning services in the post-partum follow up in prevention programs plays an important role in reducing the risk of mother to-child transmission, by reducing pregnancies among HIV-positive women. PMID- 19304423 TI - Cost-effectiveness of linezolid versus vancomycin for hospitalized patients with complicated skin and soft-tissue infections in France. AB - Studies have shown similar clinical cure rates and shorter length of hospitalization when using linezolid compared to vancomycin in patients with complicated skin and soft-tissue infections due to suspected or proven methicillin-resistant Staphylococcus aureus (MRSA). OBJECTIVE: This study had for aim to compare the cost-effectiveness of linezolid versus vancomycin in French healthcare settings. METHOD: A decision-analytic model followed an average patient from the initiation of an empiric treatment until cure, death or second line treatment failure. A clinical data probability was obtained from clinical trials, resource utilization data (including treatment duration and length of hospitalization) and prevalence of MRSA was obtained from a Delphi panel, and costs from published sources. RESULTS: First-line cure rate for linezolid-treated patients was 90.7% versus 85.5% for vancomycin; the total cure rates after two lines of treatment were 98.5% and 98.0%, respectively. The average total cost was 7,778euro for linezolid versus 8,777euro for vancomycin. The mean estimated length of hospitalization after two lines of treatment was 10.7 days for linezolid versus 13.3 days for vancomycin. The increased effectiveness and reduced cost lead to more frequent prescription. This did not change after one way sensitivity analyses. CONCLUSION: Linezolid may be considered as a cost effective treatment for patients with complicated skin and soft-tissue infections suspected to be MRSA related in France. PMID- 19304424 TI - [Spinal cord toxoplasmosis in HIV infection]. AB - We report the case of an atypical localization of a spinal cord "toxoplasmic abscess". The 46-year-old patient, HIV-1 positive, was admitted for acute urine retention and gait disorders. MRI revealed a T12-L1 medullary lesion suggesting a tumoral, inflammatory and infectious pathology. The radiological aspect and immunosuppression lead to the initiation of a treatment against Toxoplasma gondii, following the same treatment principles as for cerebral toxoplasmosis. The diagnosis can only be proved by data from autopsy or surgical biopsy, but toxoplasmosis PCR on CSF seems to be an interesting alternative to confirm the diagnosis. According to the literature, PCR is not sensitive enough as a diagnostic tool. Improvement after treatment supported the diagnosis confirmed by PCR. PMID- 19304425 TI - [Diagnosis of celiac disease in a young woman]. PMID- 19304426 TI - [Celiac disease]. PMID- 19304427 TI - [Clinical case: should esophageal cancers still be operated on?]. PMID- 19304429 TI - The influence of liquid crystal display monitors on observer performance for the detection of interstitial lung markings on both storage phosphor and flat-panel detector chest radiography. AB - PURPOSE: To compare observer performance with a flat-panel liquid crystal display (LCD) monitor and with a high-resolution gray-scale cathode ray tube (CRT) monitor in the detection of interstitial lung markings using a silicon flat-panel detector direct radiography (DR) and storage phosphor computed radiography (CR) in a clinical setting. MATERIALS AND METHODS: We displayed 39 sets of posteroanterior chest radiographs from the patients who were suspected of interstitial lung disease. Each sets consisted of DR, CR and thin-section CT as the reference standard. Image identities were masked, randomly sorted, and displayed on both five mega pixel (2048x2560x8 bits) LCD and CRT monitors. Ten radiologists independently rated their confidence in detection for the presence of linear opacities in the four fields of the lungs; right upper, left upper, right lower, and left lower quadrant. Performance of a total 6240 (39 setsx2 detector systemsx2 monitor systemx4 fieldsx10 observers) observations was analyzed by multi-reader multi-case receiver operating characteristic (ROC) analysis. Differences between monitor systems in combinations of detector systems were compared using ANOVA and paired-samples t-test. RESULTS: Area under curves (AUC) for the presence of linear opacities measured by ROC analysis was higher on the LCDs than CRTs without statistical significance (p=0.082). AUC was significantly higher on the DR systems than CR systems (p=0.006). AUC was significantly higher on the LCDs than CRTs for DR systems (p=0.039) but not different for CR systems (p=0.301). CONCLUSION: In clinical conditions, performance of the LCD monitor appears to be better for detecting interstitial lung markings when interfaced with DR systems. PMID- 19304428 TI - Molecular imaging of vessels in mouse models of disease. AB - Vascular imaging of angiogenesis in mouse models of disease requires multi modal imaging hardware capable of targeting both structure and function at different physical scales. The three dimensional (3D) structure and function vascular information allows for accurate differentiation between biological processes. For example, image analysis of vessel development in angiogenesis vs. arteriogenesis enables more accurate detection of biological variation between subjects and more robust and reliable diagnosis of disease. In the recent years a number of micro imaging modalities have emerged in the field as preferred means for this purpose. They provide 3D volumetric data suitable for analysis, quantification, validation, and visualization of results in animal models. This review highlights the capabilities of microCT, ultrasound and microPET for multimodal imaging of angiogenesis and molecular vascular targets in a mouse model of tumor angiogenesis. The basic principles of the imaging modalities are described and experimental results are presented. PMID- 19304430 TI - Determination of osthol and its metabolites in a phase I reaction system and the Caco-2 cell model by HPLC-UV and LC-MS/MS. AB - A straightforward and sensitive reversed-phase high-performance liquid chromatography (HPLC) assay was developed and validated for the analysis of osthol and its phase I metabolites (internal standard: umbelliferone). The method was validated for the determination of osthol with respect to selectivity, precision, linearity, limit of detection, recovery, and stability. The linear response range was 0.47-60 microM, and the average recoveries ranged from 98 to 101%. The inter-day and intra-day relative standard deviations were both less than 5%. Using this method, we showed that more than 80% of osthol was metabolized in 20 min in a phase I metabolic reaction system. Transport experiments in the Caco-2 cell culture model indicated that osthol was easily absorbed with high absorptive permeability (>10 x 10(-6)cm/s). The permeability did not display concentration-dependence or vectorial-dependence and is mildly temperature sensitive (activation energy less than 10 kcal/mol), indicating passive mechanism of transport. When analyzed by LC-MS/MS, five metabolites were detected in a phase I reaction system and in the receiver side of a modified Caco 2 cell model, which was supplemented with the phase I reaction system. The major metabolites appeared to be desmethyl-osthol and multiple isomers of dehydro osthol. In conclusion, a likely cause of poor osthol bioavailability is rapid phase I metabolism via the cytochrome P450 pathways. PMID- 19304431 TI - Development and validation of a liquid chromatography-tandem mass spectrometry assay for the simultaneous quantitation of prednisolone and dipyridamole in human plasma and its application in a pharmacokinetic study. AB - We have developed and validated an accurate, sensitive, and robust LC-MS/MS method that determines the concentration of CRx-102 (the combination of prednisolone and dipyridamole) in human plasma. In this method, prednisolone, dipyridamole, and the combined internal standards (IS) prednisolone-d(6) (IS for prednisolone) and dipyridamole-d(20) (IS for dipyridamole) were extracted from 100 microL human EDTA plasma using methylbutyl ether. Calibration curves were linear over a concentration range of 0.4-200 ng/mL for prednisolone and 5-3000 ng/mL for dipyridamole. The analytes were quantitatively determined using tandem mass spectrometry operated in positive electrospray ionization in a multiple reaction monitoring (MRM) mode. This validated method has been used successfully in clinical pharmacokinetic studies of CRx-102 in healthy volunteers. PMID- 19304432 TI - Analysis of conjugated steroid androgens: deconjugation, derivatisation and associated issues. AB - Gas chromatography/mass spectrometry (GC/MS) is the preferred technique for the detection of urinary steroid androgens for drug testing in athletics. Excreted in either the glucuronide or sulfated conjugated form, steroids must first undergo deconjugation followed by derivatisation to render them suitable for GC analysis. Discussed herein are the deconjugation and the derivatisation preparative options. The analytical challenges surrounding these preparatory approaches, in particular the inability to cleave the sulfate moiety have led to a focus on testing protocols that reply on glucuronide conjugates. Other approaches which alleviate the need for deconjugation and derivatisation are also highlighted. PMID- 19304433 TI - Effective on-line extraction of drugs from plasma using a restricted-access media column in column-switching HPLC equipped with a dilution system: application to the simultaneous determination of ER-118585 and its metabolites in canine plasma. AB - The present paper describes the on-line extraction of drugs in plasma using a restricted-access media (RAM) column in a column-switching high performance liquid chromatography (HPLC) apparatus that was equipped with an on-line dilution system. The use of a six- to eightfold on-line dilution ratio for plasma samples resulted in almost 100% recovery of both acidic and basic drugs from plasma. It was found that the relationship between the on-line dilution times and drug recovery efficiencies from plasma was explained in terms of the binding constant between the drug and albumin. The applicability of column-switching HPLC with an on-line dilution system and the effectiveness of the extraction procedure were confirmed by a simultaneous determination of the basic compound, ER-118585, and its metabolites in canine plasma. PMID- 19304434 TI - Hepatitis B reactivation in a hepatitis B surface antigen-negative patient after allogeneic bone marrow transplant: successful treatment with lamivudine and seroconversion. AB - Hepatitis B reactivation in hepatitis B surface antigen (HBsAg)-negative and anti HBsAg antibodies-positive patients is an infrequent complication of chemotherapy, usually with fatal evolution. Here we report an HBsAg-negative patient with a myelodysplastic syndrome, who developed hepatitis B reactivation after chemotherapy and evolved favorably after lamivudine treatment, allowing seroconversion. PMID- 19304435 TI - Strains of Burkholderia cenocepacia genomovar IIIA possessing the cblA gene that are distinct from ET12. AB - Three strains of Burkholderia cenocepacia genomovar IIIA that were polymerase chain reaction positive for cblA, bcrA, and the epidemic strain marker, but were distinct from representatives of ET12 by pulsed-field gel electrophoresis, are described. One of these strains was shown to express cable pili by electron microscopy. PMID- 19304436 TI - Histoplasma capsulatum var. duboisii infection in a patient with AIDS: rapid diagnosis using polymerase chain reaction-sequencing. AB - We describe an original case of disseminated infection with Histoplasma capsulatum (Hc) var. duboisii in an African patient with AIDS who migrated to Switzerland. The diagnosis of histoplasmosis was suggested using direct examination of tissues and confirmed in 24 h with a panfungal polymerase chain reaction assay. The variety duboisii of Hc was established using DNA sequencing of the polymorphic genomic region OLE. Molecular tools allow diagnosis of histoplasmosis in 24 h, which is drastically shorter than culture procedures. PMID- 19304437 TI - Killing of Escherichia coli by beta-lactams at different inocula. AB - Escherichia coli is a common pathogen implicated in intra-abdominal infections; a heavy bacterial burden is often encountered, and the clinical utility of beta lactams may be limited by the inoculum effect. We examined the impact of a high inoculum on the bactericidal activity of various beta-lactams against E. coli. Two wild-type, an extended-spectrum beta-lactamase-producing, and a plasmid mediated AmpC-producing strains, were used. Clinically achievable concentrations of piperacillin/tazobactam, ceftriaxone, and ertapenem were investigated. Viable bacterial burden was serially determined for 24 h by quantitative culture. All 3 beta-lactams demonstrated significant killing against the standard inoculum (10(5) CFU/mL) of susceptible strains. However, the activity of piperacillin/tazobactam was drastically reduced with 10(8) CFU/mL of bacteria. Ertapenem was the least affected by the inoculum effect in all strains. Our results suggest that different beta-lactam subclasses have a distinct killing profile against a dense E. coli population. Comparative in vivo/clinical investigations are warranted to validate our findings. PMID- 19304438 TI - Corynebacterium bovis shoulder prosthetic joint infection: the first reported case. AB - We report the first case of Corynebacterium bovis shoulder prosthetic joint infection. The organism was isolated from intraoperative tissue culture and from the removed prosthesis using sonication. A 2-stage exchange and 3 months of antibiotic therapy were performed. C. bovis may cause implant-associated infections, which can manifest as low-grade infection. PMID- 19304439 TI - Impact of liver cirrhosis on mortality in patients with community-acquired bacteremia. AB - Few studies have analyzed the impact of liver cirrhosis, a clinically significant comorbid medical condition, on the mortality of patients with community-acquired bacteremia. We conducted an observational study of 839 consecutive community acquired bacteremia patients who were hospitalized through the emergency department (ED). We compared the 30-day mortality of bacteremia patients with and without liver cirrhosis using Cox proportional hazards regression. The probability of survival at day 30 was significantly different for the cirrhotic and noncirrhotic groups (53% versus 82%, respectively; P < 0.001 by the log-rank test). Multivariate analysis indicated that liver cirrhosis was associated with an increased risk of short-term mortality (hazard ratio, 2.0; 95% confidence interval, 1.1-3.5), as well as age, higher comorbidity index, and markers obtained from clinical presentation at ED. In conclusion, in addition to the effects from other prognostic factors, liver cirrhosis has a significant impact on the mortality of patients with community-acquired bacteremia. PMID- 19304440 TI - HER-family gene amplification and expression in resected pancreatic cancer. AB - AIMS: Despite surgical resection, pancreatic cancer carries a poor prognosis. In search for new molecular therapeutic targets, we investigated the expression of the HER-family and gene amplification of HER-2 in pancreatic adenocarcinomas of different stages. METHODS: Tissue of 45 resected patients was analyzed for all HER-family 1-4 expression by immunohistochemistry and HER-2 gene amplification was assessed by multiplex ligation-dependent probe amplification and chromogenic in situ hybridization. The type of surgery, location, stage and grade of the tumor, as well as involvement of the resection margins were correlated with HER expressions and univariate and multivariate survival analysis performed. RESULTS: Normal pancreatic tissue lacked HER1-2 expression, but did show HER3-4 expression. In cancers, no membranous overexpression of HER-1 and HER-2 was seen nor gene amplification of HER-2 found. HER-3, HER-4 is physiologically expressed in the normal pancreas and loss of cytoplasmic HER-3 and HER-4 expression was seen in 33/45 (73%) and 8/45 (18%) of pancreatic cancers. Cytoplasmic HER-3 expression decreased from early to late stage (p=0.05). HER-4 expression was not associated with survival, stage or tumor grade. There were no statistically significant differences in HER1-4 expression between the papilla of Vater (n=13) and non-papilla cancers (n=32). Multivariate survival analysis showed only stage to be of independent prognostic value (p=0.015). CONCLUSIONS: HER-1 and HER-2 are not overexpressed in pancreatic cancers. HER-3 and HER-4 are expressed in the normal pancreas but expression is lost in pancreatic cancer. HER-targeted therapy in pancreatic cancer is not supported by HER-expression of the tumor. PMID- 19304441 TI - [Accidental intravenous injection of potassium chloride: analysis of contributing factors and barriers to risk reduction]. AB - Errors linked to injectable potassium chloride (KCl) have been the cause of deaths which have occurred for many years. Following an accidental direct intravenous injection of KCl of no clinical consequence for the patient, we have analyzed the contributive factors, established an action plan to prevent this risk and finally assessed its impact. Among the causes leading to medication errors, we have identified those linked to the handling of the drugs by nurses, the team, the work conditions, the organization, the institutional context and finally to the drug itself. The risk reduction procedure involved a withdrawal of injectable KCI ampoules from wards, possible in 52% of the care units, a reorganization of storage for the others. The subsequent monitoring of floorstocks revealed that these measures were insufficient and that the risks prevailed due to the presence of KCI ampoules in drawers assigned to other ionic solutions. A study carried out among the medical and nursing personnel revealed that 61.2% of the doctors thought that the risk existed in their ward and 68% of the nurses considered themselves to be exposed to the risk of a medication error. The drug supply chain of our institution, as in numerous others, is not safe. Hospitals are not yet organized adequately to prevent the occurrence of such an error. The comparison with foreign organizations of drug dispensation allows us to think that the improvement and professionalization of the drug supply chain will both be assets in the prevention of such medication errors. PMID- 19304442 TI - [Secondary hyperkalaemic acute flaccid tetraplegia]. AB - A 57-year-old man with chronic renal failure and multiple vascular diseases presented an acute flaccid tetraplegia. Blood tests revealed renal failure with serum potassium of 9.69 mmol/l. The medullar and brain MRI were normal. The correction of the hyperkalaemia led to a fast and complete neurological recovery. This fact confirmed the diagnosis of secondary hyperkalaemic tetraplegia. PMID- 19304443 TI - [Pulmonary aspiration: epidemiology and risk factors]. AB - OBJECTIVE: To determine the epidemiology and risk factors for gastric aspiration in a French university hospital. METHODS: Files were prospectively selected from a computer database of rare and severe complications between January 2002 et April 2007. Medical files were then analyzed according to a predetermined list of specific items. RESULTS: Forty patients suffered from gastric aspiration among 117 033 anaesthesias (4 medical files lost) (31/100,000 global incidence). All occurred during general anaesthesia, 83% at induction, 8% in the recovery room, half of the cases for emergent cases (15% of emergency). In emergency, the incidence of gastric aspiration increased by 4.5. Only 14 patients (39%) had a "full stomach", 17 (47%) other risk factors and five (14%) none. A rapid induction-intubation sequence with a Sellick manoeuvre was performed in only 50% of patients with a full stomach and in 23.6% of those with other risk factors. Gastric aspiration occurred in three patients with a laryngeal mask. Aspiration was associated with clinical symptoms in 21 patients. Eleven patients developed an ARDS (5 deaths). DISCUSSION: The incidence of gastric aspiration was slightly higher in this series than in other published series. Rapid induction-intubation sequence with Sellick manoeuvre is not always used in patients with a full stomach. Restraining this technique only to these latter patients does not guarantee practitioners to avoid the risk of gastric aspiration. There is a large need to define which patients are at higher risk of gastric aspiration by national recommendations of clinical practice. PMID- 19304444 TI - [Acetaminophene-induced hypotension in intensive care unit: a prospective study]. AB - OBJECTIVES: The aim of this study was to evaluate the incidence of paracetamol induced hypotension in intensive care unit (ICU). The secondary end-point was the description of pathophysiologic phenomenon during this hypotension and risk factors. STUDY DESIGN: An observational study in three ICU of a French teaching hospital. PATIENTS AND METHODS: All consecutives patients whom benefit from intravenous paracetamol administration were included in the study. When a 20% droop in arterial blood pressure occurred, plasma samples were obtained and tryptases were measured at 6 and 48 hours. Clinical, biological characteristics and paracetamol administration duration were prospectively monitored. RESULTS: During a 2-months period, 127 ICU patients were included in the study with 1507 paracetamol administration. Twenty droops in arterial blood pressure were recorded in ICU. The incidence rate was 1.33%. Administration duration was 32+/-9 min. No respiratory nor cutaneous manifestations occurred during hypotensions. A specific treatment was administrated in half of the patients. Hypotension incidence was higher (3.9%) in patients with brain injury. Eighty percent of patients with hypotension have a severe sepsis or a septic shock. CONCLUSION: In this cohort of ICU patients, hypotension incidence was higher than reported in drug legal mentions. Immunoallergic phenomenon was excluded. Brain injury and sepsis seems to be risk factors. PMID- 19304445 TI - [What are the benefits and risk of non-opioid analgesics combined with postoperative opioids?]. PMID- 19304446 TI - [National survey on out-of-hospital difficult intubation management]. AB - OBJECTIVES: Difficult intubation rate is higher in the prehospital setting than in the operating room. Goal of this survey was to assess compliance of the French prehospital mobile emergency unit (Smur) to the recent French guidelines for the difficult airway management. STUDY DESIGN: National phone survey. METHODS: A phone questionnaire was proposed to one senior emergency physician of all 380 French Smur. Seven questions were asked about intubation devices used, availability of a written difficult intubation algorithm and intubation training of the Smur's physicians. RESULTS: Guidelines of the recent French consensus conference on difficult intubation are only partly followed by the Smur. Only 60% of the Smur perform systematic rapid intubation sequence, plastic laryngoscope blades are used by more than 50% of the Smur and less than 50% of the Smur have a written difficult intubation management algorithm available. The Gum elastic Bougie is available in 58% of the Smur and the intubating laryngeal mask airway in 71%, whereas initial formation for difficult intubation devices used is provided to the emergency physicians in only 58% of the Smur. CONCLUSION: This survey shows that the French guidelines for the difficult airway management are only partially followed by the French Smur. An effort should be made for a larger diffusion of these guidelines towards the emergency physicians working in the Smur. PMID- 19304447 TI - [Impact of a quality assurance program on the use of neuromuscular monitoring and reversal of muscle relaxants]. AB - INTRODUCTION: As part of a quality assurance in the anaesthesia department, this study was designed to enhance the rate of neuromuscular blockade monitoring for patients receiving muscle relaxant during anaesthesia. METHODS: After approval of our local ethical committee, we assessed 200 computerized anaesthesia records in which neuromuscular relaxants were used. The following data were collected: demographic characteristics, durations of anaesthesia and surgery, use of neuromuscular monitoring, reversal agents and the quality of neuromuscular monitoring. The results were discussed with all anaesthesia providers of the department and an internal guideline was elaborated with the endpoint that all patients having muscle relaxants should have quantitative neuromuscular monitoring. Six months later, another assessment of 200 consecutive records collected the same data to check the efficiency of the elaborated guideline. RESULTS: The monitoring rate was of 67% at the first assessment and increased to 94% (p<0.05). The reversal rate was at 48% in the first assessment and was stable at the second assessment (50%). The rate of patients not monitored and not reversed decreased from 5 to 2% (p<0.05). DISCUSSION: This study shows that as part of a quality assurance program systematic quantitative monitoring of neuromuscular blockade can be significantly increased. PMID- 19304448 TI - [Cardiac biomarkers for diagnosis of myocardial infarction]. AB - Diagnosis of postoperative myocardial infarction is often difficult, based on tools with a low sensitivity (clinical symptoms, EKG), or with a low specifity (old biomarkers, echocardiographic abnormalities) or inadequate for clinical practice (scintigraphy). Since 1995, clinicians may use more cardiospecific markers (troponin) allowing to modify strategy for postoperative myocardial infarction diagnosis. The aim of this review is to resume such an attitude. PMID- 19304449 TI - [Management of transdermal therapeutics in perioperative care]. AB - OBJECTIVE: To provide guidance on perioperative management of most frequently encountered transdermal therapeutics in anaesthesiology: nicotine, fentanyl, nitroglycerin, scopolamine and estradiol. DATA SOURCES: A review of the last decade literature was carried out on the Pubmed database using the following keywords (transcutaneous or percutaneous or transdermal or transdermic or skin or dermal) and (drug delivery systems or therapeutic systems or drug administration) grouped under the Mesh terms cutaneous administration, perioperative care, surgery, pharmacokinetics, nicotine, fentanyl, nitroglycerin, scopolamine, estradiol. STUDY SELECTION: Original articles, general articles reviews, guidelines, letters to the editor and case reports have been selected. DATA EXTRACTION: Articles were analyzed for each transdermal treatment in terms of pharmacokinetics as well as anaesthetics and surgical interactions. DATA SYNTHESIS: Transdermal nicotine must be removed before anaesthesia of patients with coronary disease or with high risk of inhalation and in case of reconstructive surgery. Transdermal fentanyl must be maintained during the perioperative period and associated with preventive treatments of hyperalgesia. Transdermal administration of fentanyl by iontophoresis is a promising system for postoperative analgesia. Transdermal nitroglycerin must be maintained before scheduled surgery of a coronary patient. Transdermal scopolamine must be removed the day before surgery because of its side effects. It could have an interest in the prevention of postoperative nausea and vomiting, but its therapeutic method remain to be defined. Transdermal estradiol can be maintained during the perioperative period. CONCLUSION: The management of transdermal therapeutics in peri operative care can be adapted for each treatment and for each patient by knowing pharmacokinetics as well as anaesthetics and surgical interactions. In emergency situations, the actions to be taken do not generally differ, but one must be aware that the effects of trandermal treatments do not disappear immediately when removed, due to their pharmacokinetics properties. PMID- 19304450 TI - Inhibition of renin-angiotensin system attenuates periadventitial inflammation and reduces atherosclerotic lesion formation. AB - Recent evidence indicates that renin-angiotensin system (RAS) plays an important role in the pathogenesis of atherosclerosis. It was reported that inhibition of RAS with angiotensin II type 1 receptor blockers (ARBs) or angiotensin converting enzyme inhibitors (ACEIs) is effective in prevention of atherosclerosis. Here, we investigated the effects of an ARB or/and an ACEI on atherosclerosis development and periadventitial inflammation in apolipoprotein E (ApoE)-deficient mice. RT PCR revealed that major RAS components were expressed in periaortic tissue. Ang II infusion significantly increased accumulation of bone marrow derived cells into both neointima (p<0.05) and periaortic tissue (p<0.01). Male ApoE- deficient mice were treated with either vehicle, TA606A (10mg/kg/day, ARB), imidapril (3mg/kg/day, ACEI) or TA606A plus imidapril (TA606A 10mg/kg/day+imidapril 3mg/kg/day, ARB+ACEI) for 24 weeks starting at 12 weeks of age. ARB, ACEI, and ARB+ACEI significantly reduced atherosclerotic lesion formation in aorta compared with vehicle (p<0.05), with reduced expression of monocyte chemoattractant protein-1 in periaortic tissues (p<0.01). Neither blood pressure nor heart rate was changed by the treatments at these lower doses. Imidapril significantly reduced lipid deposition in atheroma and plasminogen activator inhibitor-1 expression in periadventitial tissue (p<0.05, respectively). Imidapril and combination therapy significantly attenuated macrophage infiltration into the atherosclerotic plaque (p<0.05, respectively). All treatments reduced macrophage accumulation in the periadventitial tissue 12 weeks after treatment (p<0.05, respectively). These results suggest that inhibition of renin-angiotensin system attenuates periadventitial inflammation and reduces atherosclerotic lesion formation. PMID- 19304451 TI - Relationships between social anxiety, depressive symptoms, and antisocial behaviors: evidence from a prospective study of adolescent boys. AB - Little is understood about generalized and non-generalized social anxiety disorder (SAD) and their associations with co-occurring internalizing and externalizing problems in adolescence. In the present study, we investigated adolescent boys with SAD symptoms and considered depressive symptoms as well as antisocial behaviors when looking for patterns during two developmental time periods; junior high and high school. Participants in the analyses were part of a longitudinal study. No patterns were found linking antisocial problems and non generalized SAD in either junior high or high school. Furthermore, it was uncommon for youths in the non-generalized SAD subgroup to develop comorbidity over time. The generalized SAD subgroup of boys, however, was likely to develop comorbidity either with depressive symptoms only or with depressive symptoms and antisocial tendencies. Our findings suggest that developmental pathways for SAD subgroups may differ. PMID- 19304452 TI - Maternal support for autonomy: relationships with persistence for children with Down syndrome and typically developing children. AB - Maternal behaviors and child mastery behaviors were examined in 25 children with Down syndrome and 43 typically developing children matched for mental age (24-36 months). During a shared problem-solving task, there were no group differences in maternal directiveness or support for autonomy, and mothers in the two groups used similar verbal strategies when helping their child. There were also no group differences in child mastery behaviors, measured as persistence with two optimally challenging tasks. However, the two groups differed in the relationships of maternal style with child persistence. Children with Down syndrome whose mothers were more supportive of their autonomy in the shared task displayed greater persistence when working independently on a challenging puzzle, while children of highly directive mothers displayed lower levels of persistence. For typically developing children, persistence was unrelated to maternal style, suggesting that mother behaviors may have different causes or consequences in the two groups. PMID- 19304453 TI - Ultrasound image texture analysis of the intima and media layers of the common carotid artery and its correlation with age and gender. AB - The intima-media thickness (IMT) of the common carotid artery (CCA) is widely used as an early indicator of cardiovascular disease (CVD). It was proposed but not thoroughly investigated that the composition and texture of the media layer (ML) can be used as an indicator for the risk of stroke. In this study, we investigate the application of texture analysis of the ML of the CCA and how texture is affected by age and gender. The study was performed on 100 longitudinal-section ultrasound images acquired from asymptomatic subjects at risk of atherosclerosis. The images were separated into three different age groups, namely below 50, 50-60, and above 60 years old. Furthermore, the images were separated according to gender. A total of 61 different texture features were extracted from the intima layer (IL), the ML, and the intima-media complex (IMC). The ML and the IMC were segmented manually by a neurovascular expert and also automatically by a snakes segmentation system. We have found that male patients tended to have larger media layer thickness (MLT) values as compared to the MLT of female patients of the same age. We have found significant differences among texture features extracted from the IL, ML and IMC from different age groups. Furthermore, for some texture features, we found that they follow trends that correlate with a patient's age. For example, the gray-scale median GSM of the ML falls linearly with increasing MLT and with increasing age. Our findings suggest that ultrasound image texture analysis of the media layer has potential as an assessment biomarker for the risk of stroke. PMID- 19304454 TI - An automatic method for colon segmentation in CT colonography. AB - An automatic method for the segmentation of the colonic wall is proposed for abdominal computed tomography (CT) of the cleansed and air-inflated colon. This multistage approach uses an adaptive 3D region-growing algorithm, with a self adjusting growing condition depending on local variations of the intensity at the air-tissue boundary. The method was evaluated using retrospectively collected CT scans based on visual segmentation of the colon by expert radiologists. This evaluation showed that the procedure identifies 97% of the colon segments, representing 99.8% of the colon surface, and accurately replicates the anatomical profile of the colonic wall. The parameter settings and performance of the method are relatively independent of the scanner and acquisition conditions. The method is intended for application to the computer-aided detection of polyps in CT colonography. PMID- 19304455 TI - Plant sterol intake and education level in the Spanish EPIC cohort. AB - OBJECTIVE: A cross-sectional study was conducted in the Spanish European Prospective Investigation into Cancer and Nutrition (EPIC) cohort to investigate the association among education level, as a measurement of socioeconomic position, gender, and dietary nutrient intake, focusing on plant sterols, in a Mediterranean population. METHODS: A sample of 25 615 women and 15 552 men (29-69 y old) from the Spanish EPIC cohort was recruited in 1992-1996. Nutrient and plant sterol intakes were estimated using a validated dietary history questionnaire and Spanish food composition tables. RESULTS AND CONCLUSION: Few differences in nutrient or plant sterol consumption existed between men and women with different education levels. Age and energy-adjusted linear regression models of plant sterol intake showed a small increase in subjects with lower education and higher consumption in men than in women. Homogeneity of healthy dietary habits across different socioeconomic groups in this population reflects a wide availability of characteristic Mediterranean foods at the time of recruitment. However, current changes in food supply and the increasing cost of healthy foods may lead to socioeconomic inequalities in Spain parallel to those taking place in other European populations. PMID- 19304456 TI - Where words fail, music speaks: isolated memory processes in a musical patient with schizophrenia. PMID- 19304457 TI - Family history of affective illness in schizophrenia patients: symptoms and cognition. AB - This study examined the relationship between having a family history of affective disorder and neuropsychological functioning and PANSS symptoms in schizophrenia patients falling into four exclusive family history groups (affective spectrum disorders, schizophrenia spectrum disorders, both, or neither). Schizophrenia patients with a family history of affective illness had the best performance on IQ tests and executive function measures. Symptoms showed fewer family history group differences. Schizophrenia patients with a family history of affective disorder may be a distinct subtype in the group of schizophrenias and may be biologically more similar to patients with serious affective disorder. PMID- 19304458 TI - Altered hemispheric balance of temporal cortex dopamine D(2/3) receptor binding in major depressive disorder. PMID- 19304459 TI - The Disrupted-in-Schizophrenia-1 Ser704Cys polymorphism and brain morphology in schizophrenia. AB - The Disrupted-in-Schizophrenia-1 (DISC1) polymorphism is a strong candidate for a schizophrenia-susceptibility gene as it is widely expressed in cortical and limbic regions, but the effect of its genotype variation on brain morphology in schizophrenia is not well known. This study examined the association between the DISC1 Ser704Cys polymorphism and volumetric measurements for a broad range of fronto-parietal, temporal, and limbic-paralimbic regions using magnetic resonance imaging in a Japanese sample of 33 schizophrenia patients and 29 healthy comparison subjects. The Cys carriers had significantly larger volumes of the medial superior frontal gyrus and short insular cortex than the Ser homozygotes only for healthy comparison subjects. The Cys carriers tended to have a smaller supramarginal gyrus than the Ser homozygotes in schizophrenia patients, but not in healthy comparison subjects. The right medial superior frontal gyrus volume was significantly correlated with daily dosage of antipsychotic medication in Ser homozygote schizophrenia patients. These different genotype effects of the DISC1 Ser704Cys polymorphism on the brain morphology in schizophrenia patients and healthy comparison subjects suggest that variation in the DISC1 gene might be, at least partly, involved in the neurobiology of schizophrenia. Our findings also suggest that the DISC1 genotype variation might have some relevance to the medication effect on brain morphology in schizophrenia. PMID- 19304460 TI - (17)O DOR and other solid-state NMR studies concerning the basic properties of zeolites LSX. AB - We demonstrate complementary (1)H, (17)O, (27)Al and (29)Si measurements for basic low-silica-X zeolites, which were unloaded and pyrrole and formic acid loaded. It was found that the acid-base-system is not stabile, if the loading exceeds one pyrrole molecule or two formic acid molecules per supercage.(17)O DOR NMR spectra exhibit at least four lines, which are broadened by a distribution of chemical shifts in a similar extend as the (29)Si MAS NMR spectra are broadened by distribution of Si-O-Al angles. A strong cation influence upon (17)O shifts was observed. But there was no strong influence of the acid molecules on the mean value of the (17)O shift of the spectra. PMID- 19304461 TI - Nanocomposite based on modified TiO2-BSA for functional applications. AB - A novel nanocomposite based on TiO(2)-protein for functional applications was prepared and characterized. The composite was made by covalent immobilisation of bovine serum albumin (BSA) on to the TiO(2) particles. BSA was attached to the TiO(2) powder through a three-step process. This involves the functionalization of the TiO(2) particles with silane and the subsequent coupling by glutaraldehyde to free NH(2) groups of the protein. Formation of Schiff's base was confirmed by IR spectroscopy and the optimum loading of the BSA was found to be 73.61%. Surface morphology of the composite was studied by SEM and TEM. Thermal analysis of TiO(2)-BSA composite was carried out by TGA and DSC. Structural variation of the BSA after immobilisation was studied by CD spectra. PMID- 19304462 TI - Surface and bulk characterisation of electrospun membranes: problems and improvements. AB - Electrospun membranes are used in a variety of applications, including filtration systems and sensors for chemical detection, and have attracted increased interest in the field of tissue engineering and regenerative medicine. Successful integration of these materials into a specific technology will require understanding of the fibres' surface, bulk and architectural properties. Detailed characterisation of these properties is frequently overlooked, particularly in specialised interdisciplinary fields such as tissue engineering. In this article we have reviewed the current status of the characterisation of electrospun membranes, while recommending improvements in using these techniques to better understand these very interesting nanostructured materials. PMID- 19304463 TI - [Congenital tuberculosis in preterm neonate: a case report]. AB - Congenital tuberculosis is a rare but severe disease. Diagnosis is often delayed, especially in preterm neonates. We report a premature infant born after 27 weeks of gestation and in vitro fertilization. Tuberculosis was suspected after 112 days of life in view of sepsis, respiratory distress, and the discovery of maternal tuberculosis. Mycobacterium tuberculosis was isolated in endotracheal aspirates, gastric aspirates, and stools. The infant initially received four antitubercular antibiotics over 3 months, then two antibiotics over 9 months. A wide screening for a possible nosocomial transmission from this index case was set up. At the chronological age of 2 years, the baby is healthy without after effects and no secondary cases were diagnosed. This article recalls the difficulty diagnosing congenital tuberculosis, particularly in preterm neonates. It also underlines the need to raise and eliminate the diagnosis of tuberculosis in an infertile woman. PMID- 19304464 TI - [BCG vaccine coverage in private medical practice: First data in children below two years old, seven months after the end of compulsory vaccination in France]. AB - OBJECTIVES: In July 2007, compulsory BCG vaccination for all children was replaced by a strong recommendation to vaccinate children at high risk of tuberculosis (children who live in Ile-de-France [IDF] or Guyana regions, who were born or whose parents were born in tuberculosis endemic countries, with a family history of tuberculosis or living in conditions defined as at risk by the doctor). In the absence of tools to detect an early decrease in vaccine coverage (VC) in this specific group, we conducted a survey with the main objective of measuring BCG VC in high risk children for which BCG is now recommended and who were born after the change in BCG vaccine policy. METHODS: Cross-sectional survey performed amongst physicians registered at "Infovac-France", a network of general practitioners and paediatricians particularly aware of recent changes in the field of vaccinations. Each doctor was asked to recruit, during his medical consultation, between six and 12 children aged 2-7 months (born after the end of compulsory BCG vaccination in July 2007) and 8-23 months (born after the withdrawal from the market of the multipuncture form of BCG [Monovax] in January 2006 and before the end of compulsory BCG vaccination in July 2007). Doctors were asked to fill in a structured online questionnaire. Data were standardized and analysed with Stata 9.2. RESULTS: A total of 2536 children, recruited by 279 general practitioners and paediatricians (6.5% of all contacted doctors), were included. VC in the target group of high risk children for who BCG is still recommended and who were seen by doctors working in a private medical practice was: overall 68%; 58% in children born after the end of compulsory BCG vaccination (68% in IDF, 48% outside IDF); 77% in those born after the withdrawal of Monovax from the market and before the end of compulsory BCG vaccination; 90% in children living in IDF born after the end of compulsory vaccination and considered as particularly at risk of tuberculosis (presence of vaccination criteria other than residing in IDF) and 60% in the same category of children whose sole criteria for vaccination was residing in IDF. Of doctors who worked in a private medical practice: 75% used to perform the BCG vaccination themselves and 58% had recommended or suggested vaccination to children at risk who were not yet vaccinated. Seventy-six percent of parents of children at risk of tuberculosis not yet vaccinated accepted BCG vaccination when recommended by their doctor. CONCLUSION: Our survey showed, on the one hand, insufficient VC in children seen in a private medical practice and born after the end of compulsory vaccination for whom BCG is still recommended. This should encourage the Ministry of Health to reinforce its communication concerning this new policy. On the other hand, the survey showed encouraging results concerning both the coverage of children at particularly high risk in IDF and the adherence of doctors and families to the new vaccine policy. These results should be interpreted with caution, taking into account the methodological limitations of this survey. PMID- 19304465 TI - [Cybergreedy, no more, or cyberaddict ? When teens waste their time in using computer]. AB - This paper is about the computer use by teenagers, often estimated abusive by their family circle. In most cases, it is a "simple abundant use" more than a true addiction or a passion. The author describes the psychosocial mechanisms of such behaviours and the characteristics of each one. Finally, he describes how the adults, teachers and nursing staff can face up their responsibilities to contribute towards a safe use of multimedia in young people. PMID- 19304466 TI - [Febrile seizures: current understanding of pathophysiological mechanisms]. AB - Febrile seizures (FS) are the most common etiology of seizure in infants and children. The pathophysiological mechanisms are not clearly understood. We review the current factors that are thought to be involved in FS occurrence. Genetic factors, a fever component (immune response to infection and rise in body temperature), and neurotopic viruses are discussed. There are currently no data supporting the notion that a rapid rise in temperature provokes FS. Regarding the available data, it is obvious that several factors act together to induce FS. PMID- 19304467 TI - [Intestinal fistula: an unusual complication of necrotizing enterocolitis in the preterm infant]. AB - Intestinal fistula is a rare complication in necrotizing enterocolitis (NEC) and is typically associated with a colonic stricture. We report the case of a preterm infant with severe NEC, who developed an ileocolic fistula followed by the appearance of a colonic stricture after surgical treatment. This report shows how complex the outcome of NEC can be: a contrast enema should be done in NEC when the clinical or biological outcome is not favorable, in order not to delay the diagnosis of intestinal fistula. PMID- 19304468 TI - A case of capecitabine-induced hyperpigmentation and radiation recall phenomenon. PMID- 19304469 TI - A clinical surrogate model for proof of efficacy of external phytomedicines for muscle pain. Results of a randomized, placebo-controlled clinical study in volunteers with muscle pain due to physical training. AB - PURPOSE: Surrogate models have been postulated for (re-)registration of external remedies for pain, whose active substances are in accordance to monographs. In a new human model, we investigated an ointment consisting of high dosed herbal ingredients. METHODS: We conducted a clinical study in 32 healthy volunteers. Four muscle regions were treated with the ointments (verum on the one side and placebo on the other) in a randomized and doubleblinded manner immediately after a standardized physical exercise with individual intensity, as well as after 1, 2 and 24 h. Acute muscle pain and muscle tension for each region was documented repeatedly during the following two days by visual analogue scale (VAS). Primary outcome parameter was the difference of pain during the follow up given as area under the curve (AUC) of VAS for corresponding right and left regions, treated with verum or placebo. Also the difference of muscle tension was documented and evaluated in an analogous way. RESULTS: 30 out of 32 included patients finished the study, but developed only moderate muscle pain, with highest pain scores for extension muscles of the arm. There was less pain in the course for the verum in 3 of the 4 regions, the mean difference of individual AUCs was at highest for the extension (triceps) muscles of the arm, but between-group differences failed significance. Feeling of muscle tension was higher than that of pain, with smaller mean AUCs of verum in all 4 regions; the differences were significant in total (p<0.02) and in 2 of 4 single regions. CONCLUSIONS: Physical exercise was not intensive enough to exert clear symptoms. Our volunteers with sportive background seem not to show severe symptoms of muscle pain and muscle tension even after an intensive training. Despite low levels of symptoms, verum showed better courses of muscle tension and muscle pain. For future studies it seems better not to use volunteers with sportive background but totally untrained persons in order to achieve pronounced symptoms. The model is feasible, sensitive, inexpensive and is much more clinically relevant than those, focusing on perfusion parameters of skin. PMID- 19304470 TI - Adoptive transfer of virus-specific and tumor-specific T cell immunity. AB - The adoptive transfer of T cells isolated or engineered to have specificity for diseased cells represents an ideal approach for the targeted therapy of human viral and malignant diseases. The therapeutic potential of adoptive T cell therapy for infections and cancer was demonstrated in rodent models long ago, but the task of translating this approach into an effective clinical therapy has not been easy. Carefully designed clinical trials have evaluated the transfer of antigen-specific T cells in humans, and provided insight into the barriers to efficacy and strategies to improve T cell therapy. The importance of altering the host environment to facilitate persistence and function of transferred T cells and intrinsic properties of T cells that are selected or engineered for therapy in determining their fate in vivo are key issues that have recently emerged and are informing the design of the next generation of clinical trials. PMID- 19304471 TI - Adoptive cell therapy for the treatment of patients with metastatic melanoma. AB - Adoptive cell therapy (ACT) is the best available treatment for patients with metastatic melanoma. In a recent series of three consecutive clinical trials using increasing lymphodepletion before infusion of autologous tumor infiltrating lymphocytes (TIL), objective response rates between 49% and 72% were seen. Persistence of infused cells in the circulation at one month was highly correlated with anti-tumor response as was the mean telomere length of the cells infused and the number of CD8+ CD27+ cells infused. Responses occur at all sites and appear to be durable with many patients in ongoing response beyond three years. In the most recent trial of 25 patients receiving maximum lymphodepletion, seven of the 25 patients (28%) achieved a complete response. Of the 12 patients in the three trials who achieved a complete response all but one are ongoing between 18 and 75 months. We recently demonstrated that ACT using autologous lymphocytes genetically modified to express anti-tumor T cell receptors can mediate tumor regression and this approach is now being applied to patients with common epithelial cancers. PMID- 19304472 TI - Endothelin receptor blockade in the management of pulmonary arterial hypertension: selective and dual antagonism. AB - Various treatments approved by the United States Food and Drug Administration for the management of pulmonary arterial hypertension (PAH) target three of the many pathways implicated in the development of PAH: prostacyclin-, endothelin-1 (ET-1) , and nitric oxide-mediated pathways. The objectives of this manuscript are to provide background information on the role of ET-1 in the pathogenesis of PAH, to provide theoretical considerations for the advantages and disadvantages of dual vs single endothelin receptor antagonists (ERAs) for the management of PAH, and to describe the clinical study results from randomized, double-blind, placebo controlled trials for the various ERAs. ET receptors (ET(A) and ET(B)) have different densities and distributions throughout the body and are dynamically regulated, such that blockade of ET(A) and ET(B) receptors may have different results in normal vs pathological conditions. Although differences in biological effects can be found in studies of isolated cells, blood vessels and animal models, clinical treatment studies have not identified clear differences in efficacy among the various ERAs. The main differences appear to be in safety profiles, with a greater frequency of serum liver function abnormalities occurring with the available dual ET(A)/ET(B) antagonist, and possibly higher rates of peripheral edema noted with selective ET(A) agents. Head-to-head studies will be necessary to resolve the question of whether single vs dual blockade produces better clinical results with fewer side effects in patients with PAH. PMID- 19304473 TI - Clinical predictors of the efficacy of a pulmonary rehabilitation programme in patients with COPD. AB - OBJECTIVES: To identify predictors of success for an 8week pulmonary rehabilitation programme (PRP) in patients with chronic obstructive pulmonary disease (COPD). METHODS: Sixty patients were stratified in subgroups according to baseline findings: airway obstruction (FEV(1) >/= or <50% pred), pulmonary hyperinflation (TLC > or or /= or <60mmHg). Outcome measurements of PRP were: >54m increase in 6min walking test (6MWT), or >4points reduction in total score of S. George Respiratory Questionnaire (SGRQ). Logistic regression analysis was used. RESULTS: After PRP there was a significant improvement in exercise tolerance and quality of life, which correlated with baseline FEV(1)/VC, PaO(2), SpO(2), 6MWT and SGRQ. SGRQ significantly decreased and 6MWT significantly increased after PRP in all subgroups, except for patients with CV comorbidities. Both univariate and multivariate logistic regression analyses showed that BMI>25 and resting PaO(2)<60mmHg were independent predictors of PRP efficacy in terms of improvement of 6MWT, but not of SGRQ scores. CONCLUSIONS: Clinical and functional baseline findings do not predict the response to PRP in COPD. The greater efficacy in patients with BMI>25 or with PaO(2)<60mmHg may be due to a greater deconditioning in overweight patients, and to a larger room for improvement in hypoxemic patients. PMID- 19304474 TI - Correlates of osteoporosis in chronic obstructive pulmonary disease: An underestimated systemic component. AB - RATIONALE: Chronic obstructive pulmonary disease (COPD) patients are at increased risk of osteoporosis. Osteoporosis is under diagnosed and under treated in these patients and the underlying mechanisms remain unclear. To date, screening recommendations for osteoporosis in COPD patients are not available. OBJECTIVES: To examine the prevalence of drug treatment of bone abnormalities as well as the clinical determinants of osteoporosis in COPD. METHODS: COPD patients (n=554) consecutively entering pulmonary rehabilitation were included in this cross sectional study. Medical history, current medication use, smoking status, lung function, bone mineral density, body composition and other clinical characteristics were assessed before entering pulmonary rehabilitation. Univariate- and multivariate multinomial logistic regression analyses were used to determine correlates of osteoporosis. MAIN RESULTS: Twenty-one percent of patients had osteoporosis and 41% had osteopenia. Osteoporosis was pharmacologically under treated (82% of osteoporotic patients were not receiving bone medication). Independent predictors of osteoporosis were cachexia (OR: 12.1; 95%CI: 4.5-32.7; p<0.001), age between 55 and 65 years (OR: 6.0; 95%CI: 2.2-16.3; p<0.001) and over 65 years (OR: 11.7; 95%CI: 4.1-33.1; p=<0.001). Overweight (OR: 0.1; 95%CI: 0.05-0.4; p=0.001) and obesity (OR: 0.78; 95%CI: 0.02-0.4; p=0.002) showed a substantial protective effect. CONCLUSIONS: The majority of COPD patients with osteoporosis entering pulmonary rehabilitation did not receive pharmacological treatment for osteoporosis. Cachectic COPD patients should be screened for osteoporosis, especially when over 55 years of age. PMID- 19304475 TI - Ascertainment of collagen vascular disease in patients presenting with interstitial lung disease. AB - INTRODUCTION: Previous studies of interstitial lung disease (ILD) suggest that prognosis and therapeutic response are influenced by the presence of underlying collagen vascular disease (CVD). Yet, what proportion of patients presenting with ILD have CVD is largely unknown. We sought to determine the frequency of a new CVD diagnosis in an ILD referral population. MATERIALS/PATIENTS AND METHODS: We retrospectively studied 114 consecutive patients evaluated at the Johns Hopkins Interstitial Lung Disease Clinic for the development of CVD. RESULTS: In this retrospective cohort, nearly one-third of the 114 patients with confirmed ILD satisfied published criteria for a CVD diagnosis. Seventeen (15%) patients were diagnosed with a new CVD as a direct consequence of their ILD evaluation. Patients with new CVD diagnosis were younger than those without new CVD diagnosis: 51.4years (95% CI 45-58years) and 60years (95% CI 57-63), respectively (p=0.01). Moreover, an ANA>or=1:640 (p=0.03) and elevated levels of creatine phosphokinase (CPK) or aldolase (p<0.001) were associated with a new CVD diagnosis. CONCLUSIONS: Unrecognized collagen vascular disease may be more common than previously appreciated among patients referred with ILD. High titer ANA and an elevated CPK or aldolase are associated with a CVD diagnosis in this referral population. PMID- 19304476 TI - Stochastic population model for electrical stimulation of the auditory nerve. AB - We have developed a biophysical model of a population of electrically stimulated auditory nerve fibers. It can be used to interpret results from physiological and behavioral experiments with cochlear implants and propose novel stimulation strategies. Our model consists of myelinated internodes described by a passive resistor-capacitor network, membrane capacitance, and leakage current at the nodes of Ranvier, as well as stochastic representations of nodal voltage dependent channels. To approximate physiological properties measured in the auditory nerve (AN) of an acutely deafened cat, electrical parameters of the model fiber were chosen based on literature-reported values. Using our model, we have replicated the following properties within 10 % of the reported feline single-fiber measurements: relative spread (5.8 %), spike latency (630 micros), jitter (93 micros), chronaxie (238 micros), relative refractory period (4.6 ms), and conduction velocity (14 m/s). Moreover, we have successfully matched response characteristics of a population of fibers with the same number of diameter distributed model fibers, enabling us to simulate responses of the entire AN. To demonstrate the performance of our model, we compare responses of a population of ANs stimulated with two speech encoding strategies, Continuous Interleaved Sampling and Compressed Analog. PMID- 19304477 TI - Characterizing response to elemental unit of acoustic imaging noise: an FMRI study. AB - Acoustic imaging noise produced during functional magnetic resonance imaging (fMRI) studies can hinder auditory fMRI research analysis by altering the properties of the acquired time-series data. Acoustic imaging noise can be especially confounding when estimating the time course of the hemodynamic response (HDR) in auditory event-related fMRI (fMRI) experiments. This study is motivated by the desire to establish a baseline function that can serve not only as a comparison to other quantities of acoustic imaging noise for determining how detrimental is one's experimental noise, but also as a foundation for a model that compensates for the response to acoustic imaging noise. Therefore, the amplitude and spatial extent of the HDR to the elemental unit of acoustic imaging noise (i.e., a single ping) associated with echoplanar acquisition were characterized and modeled. Results from this fMRI study at 1.5 T indicate that the group-averaged HDR in left and right auditory cortex to acoustic imaging noise (duration of 46 ms) has an estimated peak magnitude of 0.29% (right) to 0.48% (left) signal change from baseline, peaks between 3 and 5 s after stimulus presentation, and returns to baseline and remains within the noise range approximately 8 s after stimulus presentation. PMID- 19304478 TI - Coupling poroelasticity and CFD for cerebrospinal fluid hydrodynamics. AB - This research uses a novel coupling of poroelastic theory and computational fluid dynamics to investigate acute hydrocephalus resulting from stenosis of the cerebral aqueduct. By coupling poroelastic theory with a multidimensional simulation of the cerebral aqueduct we are able to investigate, for the first time, the impact of physically relevant stenosis patterns on ventricular enlargement, accounting for the nonintuitive long time history responses of the ventricular system. Preliminary findings demonstrate clearly the importance that the fluidic-poroelastic coupling plays: ventricular enlargement is significantly smaller with local stenosis patterns and almost all of the observable pressure drop occurs across the stenosis. Short timescale effects [O(heartbeat)] are explored and their contribution to the long timescales interrogated. PMID- 19304479 TI - Metabolic prosthesis for oxygenation of ischemic tissue. AB - This communication discloses new ideas and preliminary results on the development of a metabolic prosthesis for local oxygenation of ischemic tissue under physiologically neutral conditions. We report for the first time selective electrolysis of physiological saline by repetitively pulsed, charge-limited electrolysis for the production of oxygen and suppression of free chlorine. Using 800-microA amplitude current pulses and < 200 micros pulse duration, we demonstrate prompt oxygen production and delayed chlorine production at the surface of a fused 0.85-mm diameter spherical platinum electrode. The data, interpreted in terms of the ionic structure of the electric double layer, suggest a strategy for in situ production of metabolic oxygen via a new class of "smart" prosthetic implants for ischemic disease such as diabetic retinopathy. We also present data indicating that collateral pH drift, if any, can be held constant using a feedback-controlled three-electrode electrolysis system that chooses an anode and cathode pair based on pH data provided by a local sensor. PMID- 19304480 TI - A novel chaotic neural network with the ability to characterize local features and its application. AB - To provide an ability to characterize local features for the chaotic neural network (CNN), Gauss wavelet is used for the self-feedback of the CNN with the dilation parameter acting as the bifurcation parameter. The exponentially decaying dilation parameter and the chaotically varying translation parameter not only govern the wavelet self-feedback transform but also enable the CNN to generate complex dynamics behavior preventing the network from being trapped in the local minima. Analysis of the energy function of the CNN indicates that the local characterization ability of the proposed CNN is effectively provided by the wavelet self-feedback in the manner of inverse wavelet transform and that the proposed CNN can achieve asymptotical stability. The experimental results on traveling salesman problem (TSP) suggest that the proposed CNN has a higher average success rate for obtaining globally optimal or near-optimal solutions. PMID- 19304481 TI - Trace ratio problem revisited. AB - Dimensionality reduction is an important issue in many machine learning and pattern recognition applications, and the trace ratio (TR) problem is an optimization problem involved in many dimensionality reduction algorithms. Conventionally, the solution is approximated via generalized eigenvalue decomposition due to the difficulty of the original problem. However, prior works have indicated that it is more reasonable to solve it directly than via the conventional way. In this brief, we propose a theoretical overview of the global optimum solution to the TR problem via the equivalent trace difference problem. Eigenvalue perturbation theory is introduced to derive an efficient algorithm based on the Newton-Raphson method. Theoretical issues on the convergence and efficiency of our algorithm compared with prior literature are proposed, and are further supported by extensive empirical results. PMID- 19304482 TI - A novel blood glucose regulation using TSK0-FCMAC: a fuzzy CMAC based on the zero ordered TSK fuzzy inference scheme. AB - This paper presents a novel blood glucose regulation for type I (insulin dependent) diabetes mellitus patients using biologically inspired TSK0-FCMAC, a fuzzy cerebellar model articulation controller (CMAC) based on the zero-ordered Takagi-Sugeno-Kang (TSK) fuzzy inference scheme. TSK0 -FCMAC is capable of performing localized online training with an effective fuzzy inference scheme that could respond swiftly to changing environment such as human's endocrine system. Without prior knowledge of disturbance (e.g., food intake), the proposed fuzzy CMAC is able to capture the glucose-insulin dynamics of individuals under different dietary profiles. Preliminary simulations show that the blood glucose level is kept within the state of euglycemia. The design of the proposed system follows closely to what is available in real life and is suitable for animal and clinical pilot testing in the near future. PMID- 19304483 TI - An entropy interpretation of the logarithmic image processing model with application to contrast enhancement. AB - The logarithmic image processing (LIP) model is a mathematical theory that provides new operations for image processing. The contrast definition has been shown to be consistent with some important physical laws and characteristics of human visual system. In this paper, we establish an information-theoretic interpretation of the contrast definition. We show that it can be expressed as a combination of the relative entropy and Shannon's information content. Based on this new interpretation, we propose an adaptive algorithm for enhancing the contrast and sharpness of noisy images. PMID- 19304484 TI - Phase-adaptive superresolution of mammographic images using complex wavelets. AB - This correspondence describes a new superresolution approach for enhancing the resolution of mammographic images using complex wavelet frequency information. This method allows regions of interest of a mammographic image to be viewed in enhanced resolution while reducing the patient exposure to radiation. The proposed method exploits the structural characteristics of breast tissues being imaged and produces higher resolution mammographic images with sufficient visual fidelity that fine image details can be discriminated more easily. In our approach, the superresolution problem is formulated as a constrained optimization problem using a third-order Markov prior model and adapts the priors based on the phase variations of the low-resolution mammographic images. Experimental results indicate the proposed method is more effective at preserving the visual information when compared with existing resolution enhancement methods. PMID- 19304485 TI - Radon representation-based feature descriptor for texture classification. AB - In this paper, we aim to handle the intraclass variation resulting from the geometric transformation and the illumination change for more robust texture classification. To this end, we propose a novel feature descriptor called Radon representation-based feature descriptor (RRFD). RRFD converts the original pixel represented images into Radon-pixel images by using the Radon transform. The new Radon-pixel representation is more informative in geometry and has a much lower dimension. Subsequently, RRFD efficiently achieves affine invariance by projecting an image (or an image patch) from the space of Radon-pixel pairs onto an invariant feature space by using a ratiogram, i.e., the histogram of ratios between the areas of triangle pairs. The illumination invariance is also achieved by defining an illumination invariant distance metric on the invariant feature space. Comparing to the existing Radon transform-based texture features, which only achieve rotation and/or scaling invariance, RRFD achieves affine invariance. The experimental results on CUReT show that RRFD is a powerful feature descriptor that is suitable for texture classification. PMID- 19304486 TI - Epileptic seizure detection in EEGs using time-frequency analysis. AB - The detection of recorded epileptic seizure activity in EEG segments is crucial for the localization and classification of epileptic seizures. However, since seizure evolution is typically a dynamic and nonstationary process and the signals are composed of multiple frequencies, visual and conventional frequency based methods have limited application. In this paper, we demonstrate the suitability of the time-frequency (t-f) analysis to classify EEG segments for epileptic seizures, and we compare several methods for t-f analysis of EEGs. Short-time Fourier transform and several t-f distributions are used to calculate the power spectrum density (PSD) of each segment. The analysis is performed in three stages: 1) t-f analysis and calculation of the PSD of each EEG segment; 2) feature extraction, measuring the signal segment fractional energy on specific t f windows; and 3) classification of the EEG segment (existence of epileptic seizure or not), using artificial neural networks. The methods are evaluated using three classification problems obtained from a benchmark EEG dataset, and qualitative and quantitative results are presented. PMID- 19304487 TI - Overview of advanced computer vision systems for skin lesions characterization. AB - During the last years, computer-vision-based diagnosis systems have been used in several hospitals and dermatology clinics, aiming mostly at the early detection of skin cancer, and more specifically, the recognition of malignant melanoma tumour. In this paper, we review the state of the art in such systems by first presenting the installation, the visual features used for skin lesion classification, and the methods for defining them. Then, we describe how to extract these features through digital image processing methods, i.e., segmentation, border detection, and color and texture processing, and we present the most prominent techniques for skin lesion classification. The paper reports the statistics and the results of the most important implementations that exist in the literature, while it compares the performance of several classifiers on the specific skin lesion diagnostic problem and discusses the corresponding findings. PMID- 19304488 TI - Automatic detection of bioabsorbable coronary stents in IVUS images using a cascade of classifiers. AB - Bioabsorbable drug-eluting coronary stents present a very promising improvement to the common metallic ones solving some of the most important problems of stent implantation: the late restenosis. These stents made of poly-L-lactic acid cause a very subtle acoustic shadow (compared to the metallic ones) making difficult the automatic detection and measurements in images. In this paper, we propose a novel approach based on a cascade of GentleBoost classifiers to detect the stent struts using structural features to code the information of the different subregions of the struts. A stochastic gradient descent method is applied to optimize the overall performance of the detector. Validation results of struts detection are very encouraging with an average F-measure of 81%. PMID- 19304489 TI - Finding multiple coherent biclusters in microarray data using variable string length multiobjective genetic algorithm. AB - Microarray technology enables the simultaneous monitoring of the expression pattern of a huge number of genes across different experimental conditions. Biclustering in microarray data is an important technique that discovers a group of genes that are coregulated in a subset of conditions. Biclustering algorithms require to identify coherent and nontrivial biclusters, i.e., the biclusters should have low mean squared residue and high row variance. A multiobjective genetic biclustering technique is proposed here that optimizes these objectives simultaneously. A novel encoding scheme that uses variable chromosome length is developed. Moreover, a new quantitative measure to evaluate the goodness of the biclusters is proposed. The performance of the proposed algorithm has been evaluated on both simulated and real-life gene expression datasets, and compared with some other well-known biclustering techniques. PMID- 19304490 TI - Rough sets and near sets in medical imaging: a review. AB - This paper presents a review of the current literature on rough-set- and near-set based approaches to solving various problems in medical imaging such as medical image segmentation, object extraction, and image classification. Rough set frameworks hybridized with other computational intelligence technologies that include neural networks, particle swarm optimization, support vector machines, and fuzzy sets are also presented. In addition, a brief introduction to near sets and near images with an application to MRI images is given. Near sets offer a generalization of traditional rough set theory and a promising approach to solving the medical image correspondence problem as well as an approach to classifying perceptual objects by means of features in solving medical imaging problems. Other generalizations of rough sets such as neighborhood systems, shadowed sets, and tolerance spaces are also briefly considered in solving a variety of medical imaging problems. Challenges to be addressed and future directions of research are identified and an extensive bibliography is also included. PMID- 19304491 TI - Dimensionality reduction oriented toward the feature visualization for ischemia detection. AB - An effective data representation methodology on high-dimension feature spaces is presented, which allows a better interpretation of subjacent physiological phenomena (namely, cardiac behavior related to cardiovascular diseases), and is based on search criteria over a feature set resulting in an increase in the detection capability of ischemic pathologies, but also connecting these features with the physiologic representation of the ECG. The proposed dimension reduction scheme consists of three levels: projection, interpretation, and visualization. First, a hybrid algorithm is described that projects the multidimensional data to a lower dimension space, gathering the features that contribute similarly in the meaning of the covariance reconstruction in order to find information of clinical relevance over the initial training space. Next, an algorithm of variable selection is provided that further reduces the dimension, taking into account only the variables that offer greater class separability, and finally, the selected feature set is projected to a 2-D space in order to verify the performance of the suggested dimension reduction algorithm in terms of the discrimination capability for ischemia detection. The ECG recordings used in this study are from the European ST-T database and from the Universidad Nacional de Colombia database. In both cases, over 99% feature reduction was obtained, and classification precision was over 99% using a five-nearest-neighbor classifier (5 NN). PMID- 19304492 TI - An interoperability test framework for HL7-based systems. AB - Health Level Seven (HL7) is a prominent messaging standard in the eHealth domain, and with HL7 v2, it addresses only the messaging layer. However, HL7 implementations also deal with the other layers of interoperability, namely the business process layer and the communication layer. This need is addressed in HL7 v3 by providing a number of normative transport specification profiles. Furthermore, there are storyboards describing HL7 v3 message choreographies between specific roles in specific events. Having alternative transport protocols and descriptive message choreographies introduces great flexibility in implementing HL7 standards, yet, this brings in the need for test frameworks that can accommodate different protocols and permit the dynamic definition of test scenarios. In this paper, we describe a complete test execution framework for HL7 based systems that provides high-level constructs allowing dynamic set up of test scenarios involving all the layers in the interoperability stack. The computer interpretable test description language developed offers a configurable system with pluggable adaptors. The Web-based GUIs make it possible to test systems over the Web anytime, anywhere, and with any party willing to do so. PMID- 19304493 TI - A novel finite-element-based algorithm for fluorescence molecular tomography of heterogeneous media. AB - The knowledge of optical properties distribution of heterogeneous media has significant impact on the reconstructed fluorescence image quality in fluorescence molecular tomography (FMT). In this study, a novel finite-element based algorithm for FMT of heterogeneous media is proposed. In the algorithm, optical properties are reconstructed using the conjugate gradient method. A modified method based on reverse differential scheme is deduced for calculating the gradient when the detector points are not restricted on boundary nodes. With the recovered optical properties, linear relationship between known surface measurements of emission light and unknown fluorescence yield is then obtained. FMT reconstruction is implemented by combining algebraic reconstruction technique (ART) and Landweber iteration method. With initial value provided by ART, Landweber iteration method improves the quantification smoothly with small step length between neighboring iterations. The algorithm was evaluated using phantoms of different heterogeneity configurations. Results show that the reconstructed fluorescence yield is insensitive to various degrees of heterogeneity for the proposed algorithm. In contrast, when assuming homogeneous optical properties, it shows that more underestimation of optical properties results in more underestimation of the reconstructed fluorescence yield. Fast computation speed of the proposed algorithm is also demonstrated in this study. PMID- 19304494 TI - Neck and face surface electromyography for prosthetic voice control after total laryngectomy. AB - The electrolarynx (EL) is a common rehabilitative speech aid for individuals who have undergone total laryngectomy, but they typically lack pitch control and require the exclusive use of one hand. The viability of using neck and face surface electromyography (sEMG) to control the onset, offset, and pitch of an EMG controlled EL (EMG-EL) was studied. Eight individuals who had undergone total laryngectomy produced serial and running speech using a typical handheld EL and the EMG-EL while attending to real-time visual sEMG biofeedback. Running speech tokens produced with the EMG-EL were examined for naturalness by 10 listeners relative to those produced with a typical EL using a visual analog scale. Serial speech performance was assessed as the percentage of words that were fully voiced and pauses that were successfully produced. Results of the visual analog scale assessment indicated that individuals were able to use the EMG-EL without training to produce running speech perceived as natural as that produced with a typical handheld EL. All participants were able to produce running and serial speech with the EMG-EL controlled by sEMG from multiple recording locations, with the superior ventral neck or submental surface locations providing at least one of the two best control locations. PMID- 19304495 TI - Semiconductor quantum dots for biosensing and in vivo imaging. AB - Semiconductor quantum dots (QDs) have captivated researchers in the biomedical field over the last decade. Compared to organic dyes and fluorescent proteins, QDs have unique optical properties such as tunable emission spectra, improved brightness, superior photostability, and simultaneous excitation of multiple fluorescence colors. Since the first successful reports on the biological use of QDs a decade ago, QDs and their bioconjugates have been successfully applied to various imaging applications including fixed cell labeling, live-cell imaging, in situ tissue profiling, fluorescence detection and sensing, and in vivo animal imaging. In this review, we will briefly survey the optical properties of QDs, the biofunctionalization strategies, and focus on their biosensing and in vivo imaging applications. We conclude with a discussion on the issues and perspectives on QDs as biosensing probes and in vivo imaging agents. PMID- 19304496 TI - Chloroform- and water-soluble sol-gel derived Eu+++/Y2O3 (red) and Tb+++/Y2O3 (green) nanophosphors: synthesis, characterization, and surface modification. AB - Eu+++ and Tb+++ ions have been incorporated into nanodimensional yttrium oxide host matrices via a sol-gel process using Y5O(OPr(i))13 as precursor (OPr(i) = isopropoxy). The as-synthesized white powders have been annealed at different temperatures. Photoluminescence (PL) spectroscopy and X-ray diffraction (XRD) have been used as tools for documenting the characteristics of these powders. For Eu+++-doped powders, a comparison of the Eu+++, 5D0-->7F1, and 5D0-->7F2 peak intensities in the emission spectra reveals that the dopant ions are occupying unsymmetrical sites in the host yttrium oxide in all the samples. For Tb+++-doped powders, the characteristic terbium 5D3-->7Fn and 5D-->7Fn (n = 2-6) transitions were visible only in the samples that had been annealed above 500 degrees C. Samples of the doped particle powders were suspended in chloroform by fragmenting the powder with and without sonification under the presence of trioctylphosphine oxide, or a mixture of oleic acid and dioctyl ether. The resulting clear colorless (for Eu+++) and light green translucent (for Tb+++) solutions of the suspended particles showed red and green luminescence upon UV excitation, respectively. In addition, suspension in water has been achieved by fragmenting the powder in the presence of dichloroacetic acid. Transmission electron micrograph investigation of the soluble particles shows single dispersed particles along with agglomerates. The changes in the luminescence due to fragmentation of the particle powder and due the influence of the surfactant of the suspended colloidal particles are discussed. PMID- 19304497 TI - Toxicity of CdTe quantum dots in bacterial strains. AB - Contradictory results on quantum dot cytotoxicity exist for many types of biological systems, especially microorganisms. In this study, we compare the cytotoxicity of CdTe quantum dots (QDs) to four very different environmental bacterial strains, giving quantitative models of the growth curves for exposed organisms. The mechanisms of toxicity are explored by measuring reactive oxygen species generation by the QDs alone and investigating the oxidative damage to mutant bacteria especially sensitive to ROS. Electron microscopic examination also reveals factors that may contribute to resistance to nanoparticles in some strains. PMID- 19304498 TI - Fabrication and characterization of silk-fibroin-coated quantum dots. AB - We report a novel technique of directly coating colloidal CdSe/ZnS core/shell quantum dots (QDs) with silk fibroin (SF), a protein derived from the Bombyx mori silk worm. The approach results in protein-modified QDs with little or no particle aggregation, and mitigates the issue of biocompatibility. QDs have desirable optical properties, such as narrow-band emission, broadband absorption, high quantum yield, and high resistance to photobleaching. SF is a fibrous protein polymer with a biomimetic peptide sequence, water and oxygen permeability, low inflammatory response, no thrombogenecity, and cellular biocompatibility, which are desirable properties for in vivo delivery. Combining the unique properties of QDs with the biocompatibility profile of SF, the approach produces particles representing a powerful tool for numerous in vivo and in vitro applications. The design and preparation of these protein-modified QDs conjugates is reported along with functional characterization using luminescence, transmission electron microscope (TEM), and atomic force microscope (AFM). Additionally, we report results obtained using the QDs conjugates as a fluorescent label for bioimaging HEYA8 ovarian cancer cells. PMID- 19304499 TI - Magnetically responsive nanoparticles for drug delivery applications using low magnetic field strengths. AB - The purpose of this study is to investigate the potential of magnetic nanoparticles for enhancing drug delivery using a low oscillating magnetic field (OMF) strength. We investigated the ability of magnetic nanoparticles to cause disruption of a viscous biopolymer barrier to drug delivery and the potential to induce triggered release of drug conjugated to the surfaces of these particles. Various magnetic nanoparticles were screened for thermal response under a 295-kHz OMF with an amplitude of 3.1 kA/m. Based on thermal activity of particles screened, we selected the nanoparticles that displayed desired characteristics for evaluation in a simplified model of an extracellular barrier to drug delivery, using lambda DNA/HindIII. Results indicate that nanoparticles could be used to induce DNA breakage to enhance local diffusion of drugs, despite low temperatures of heating. Additional studies showed increased diffusion of quantum dots in this model by single-particle tracking methods. Bimane was conjugated to the surface of magnetic nanoparticles. Fluorescence and transmission electron microscope images of the conjugated nanoparticles indicated little change in the overall appearance of the nanoparticles. A release study showed greater drug release using OMF, while maintaining low bulk heating of the samples (T = 30 degrees C). This study indicates that lower magnetic field strengths may be successfully utilized for drug delivery applications as a method for drug delivery transport enhancement and drug release switches. PMID- 19304500 TI - Nanoscintillators for microscopic diagnostics of biological and medical objects and medical therapy. AB - The main focus of this paper is the description of qualitatively new facilities for diagnostics of biological and medical objects and medical therapy obtained by applications of nanocrystalline scintillators. These facilities are based on abilities of nanoscintillators to selective conjugation with various biomolecular objects and noticeable variations of their atomic structures, X-ray diffraction (XRD) patterns, and light-emission characteristics induced by modifications of conditions on their external surfaces. Experimental results presented in this paper provide development of detection in vivo just inside a living organism of various viruses, cancer cells, and other pathological macromolecules by means of scanning X-ray diffractometry of nanoparticles introduced into the body. These data are produced by selective adsorption of pathological bioobjects by these nanoparticles and subsequent modifications of their XRD patterns. Application of narrow collimated X-ray beams and new types of X-ray detector matrices providing microscopic spatial resolution due to usage of nanoscintillators enables determination of the regions where these pathologies are localized with high accuracy. The procedure of detection of pathological organelles by this method improves possibilities for effective destruction of these pathologies by low-dose X-ray irradiation of the places of their localization. High effectiveness of this X-ray destruction is provided by concentrated absorption of X-ray quanta by the nanoscintillators and direct transfer of the absorbed energy to the pathological objects that are attached to the absorbing particles. Constructions of 3-D radiation detector matrices providing necessary microscopic spatial and angular resolutions of X-ray imaging are described on the basis of nanoscintillators, fiber light guides, and microcapillary matrices. PMID- 19304501 TI - Immune response induced by fluorescent nanocrystal quantum dots in vitro and in vivo. AB - Fluorescent nanocrystal quantum dots (QDs) are widely used as novel tools in various biological fields including cellular biology, molecular biology, and even in basic and clinical medical fields, due to their far brighter photoemission and photostability. Although many amounts of biological studies, including in vivo experiments, were circumstantially investigated, there is no informative report that investigates whether the QDs affect the mammalian immune system. This study investigated the immune response and biological behavior of QDs in vitro and in vivo. The immune response to QDs by both lymphocytes and kinds of macrophages in vitro and in vivo was investigated. Co-culture of QDs with immune cells showed that apparently normal production of cytokines and chemokines in both mouse CD4+ lymphocytes and peritoneal F4/80+ macrophages (PM phi). In addition, the bionanocomplex of QDs with enhanced-green-fluorescent-protein (eGFP)-encoding nucleotides successfully induced the expression of eGFP protein by PM phi. However, direct injection of QD+nucleotides bionanocomplex aqueous solution into the peritoneal cavity of mice resulted in the inflammation with the infiltration of inflammatory cells into the peritoneal cavity. Furthermore, QD+nucleotides bionanocomplex (but not QD bionanocomplex without nucleotides), induced the production of both proinflammatory cytokines and chemokines by PM phi in vitro. These results indicated that QDs covered with nucleotides caused the peritoneal inflammation in vivo via activation of PM phi and probably nonimmune cells. Taken together, these data indicated that QDs affect the proliferation of immune cells, but not in immune response including cytokine production. We propose here that all nanotechnology researchers should confirm the biological responses of their nanoscale products, because the biological response against nanoscale products can be occurred by not only in immune cells but also other nonimmune cells. PMID- 19304502 TI - Synthesis and characterization of double-layer quantum-dots-tagged microspheres. AB - Quantum-dots-tagged poly (styrene-acrylamide-acrylic acid) microspheres (QDsAAMs) were synthesized and modified with hydrazine hydrate through hydrazinolysis. Azidocarbonyl groups, which can be rapidly coupled with proteins under mild conditions, were introduced onto the surface of QDsAAM using azido reaction. Bovine serum albumin (BSA) was selected as model protein to be covalently immobilized on the azidocarbonyl QDsAAM. Instruments such as fluorescence microscope, optical microscope, confocal laser scanning microscope, UV-visible spectrometer, Fourier transform infrared spectrometer, size analyzer, and fluorescence spectrophotometer were used to characterize QDsAAM. Results showed that QDsAAM had a regular double-layer spherical shape and an average diameter of 11.2 microm. It also displayed high fluorescence intensity (lambda(ex)/lambda(em) = 250 nm/370 nm), which showed linearity with concentrations ranging from 3.0 x10(-3) to 90.0 x10(-3) g.L(-1). In addition, external factors such as pH and ionic strength exerted little influence on fluorescent characteristic. BSA immobilization indicated that QDsAAM with azidocarbonyl groups could be covalently coupled with BSA at the rate of 40 x10(-3) g/g (BSA/QDsAAM), while fluorescence linearity correlation was also found. This functional azidocarbonyl QDsAAM with sensitive fluorescence and active azidocarbonyl groups could be used as a promising fluorescent probe for quantitative detection, protein immobilization, and early rapid clinical diagnostics. PMID- 19304503 TI - Tumor-targeted quantum dots can help surgeons find tumor boundaries. AB - Despite surgical advances and recent progress in adjuvant therapies, the prognosis for patients with malignant brain tumors such as glioblastoma multiforme has remained poor, and the neurological deterioration suffered by most patients as a consequence of tumor progression is dramatic and severe. In addition, malignant brain tumors have >>95% recurrence close to the primary site of initial resection. Unfortunately, standard imaging techniques do not permit the intraoperative identification of individual or small clusters of residual tumor cells, precluding their selective removal while sparing the surrounding normal brain tissue. In this report, we show that quantum dots (QDs) coupled to epidermal growth factor (EGF) or anti-EGF receptor receptor (EGFR, Her1) specifically and sensitively label glial tumor cells in cell culture, glioma mouse models, and human brain-tumor biopsies. A clear demarcation between brain and tumor tissue at the macroscopic as well as the cellular level is provided by the fluorescence emission of the QDs. PMID- 19304504 TI - Electrophysiological characterisation of myoclonic-atonic seizures in symptomatic continuous spike-waves during slow sleep syndrome. AB - Sudden epileptic falls are frequently reported in continuous spike-waves during slow sleep (CSWS) syndrome. Inhibitory seizures are usually considered as the underlying mechanism. However, published polygraphic recordings are rare. We report the case of a 22 month-old boy suffering from a symptomatic CSWS syndrome associated with a perinatal stroke involving the right middle cerebral artery territory. He presented with psychomotor regression and daily multiple falls related to myoclonic-atonic seizures. Neurophysiological examination showed secondary generalized myoclonus systematically correlated with a bilateral spike spreading from the right central area. This confirms that positive myoclonus, in addition to negative myoclonus, may be responsible for epileptic falls in CSWS syndrome. [Published with video sequences]. PMID- 19304505 TI - [Single nucleotide polymorphisms of DAT and DRD(2) genes in Han Chinese population and their association with stuttering]. AB - OBJECTIVE: To explore the correlations of dopamine transporter gene (DAT) and dopamine D(2) receptor gene (DRD2) to stuttering. METHODS: To examine the correlations of the 5 single nucleotide polymorphisms (SNPs) in dopaminergic gene (C252T, C1804T, and C1820T in DAT gene, and T1054C and C1072T in DRD(2) gene) to stuttering in Han Chinese individuals, a case-control study involving 112 patients with stuttering and 112 gender-matched controls was carried out. Genotyping was performed by a combined approach using polymerase chain reaction (PCR) and pyrosequencing. RESULTS: C1804T showed no polymorphism in either the patients or the control subjects and was therefore excluded from the following analysis. The C allele frequency at C1072T site was significantly higher, but T allele frequency significantly lower in the stuttering group than in the control group. The patients had significantly higher CC and lower CT genotype frequencies than the control group. There were no significant differences in the allelic frequencies of C252T, C1820T and T1054C between the patients and the controls, suggesting a Hardy-Weinberg equilibrium at these 3 loci. CONCLUSION: The presence of the C allele at C1072T in DRD(2) gene is associated with increased susceptibility to stuttering in Han Chinese, whereas the T allele provides protection against the onset of stuttering. PMID- 19304506 TI - [Prokaryotic expression of S2 extracellular domain of SARS coronavirus spike protein and its fusion with Hela cell membrane]. AB - OBJECTIVE: To construct the expression plasmid of S2 extracellular domain (S2ED) of SARS-coronavirus (SARS- Cov) spike protein (S protein) and enhanced green fluorescent protein (EGFP) to obtain the fusion protein expressed in prokaryotic cells. METHODS: S2ED based on bioinformatics prediction and EGFP sequence were amplified by PCR and inserted into pET-14b plasmid. The recombinant protein His S2ED-EGFP was expressed in E. coli by IPTG induction. After purification by Ni NTA agarose beads, the soluble fractions of the fusion protein were collected and identified by SDS-PAGE and Western blotting. The fusion of S2ED with Hela cell membranes was observed with fluorescent microscope. RESULTS: The pET-14b-S2ED EGFP plasmid was correctly constructed and highly expressed in BL21 (DE3). When incubated with Hela cells, the purified protein could not internalize through membrane fusion. CONCLUSIONS: The expression plasmid containing S2ED of SARS-Cov S protein and EGFP sequence is constructed successfully. Although the recombinant protein obtained has not shown the expected fusion effect with Hela cell membrane, this work may enrich the understanding of the process of membrane fusion mediated by S2 protein and lay the foundation for future study of targeting cell transport system based on cell-specific binding peptide. PMID- 19304507 TI - [Full-length of human papillomavirus type 18L1 gene optimized using the plant preferred codons and synthesized by overlapping PCR]. AB - OBJECTIVE: The Human papillomavirus type 18L1 (HPV18L1) gene was synthesized by overlapping PCR after optimization using plant preferred codons. METHODS: The gene sequences of HPV18L1 were obtained from GenBank and analyzed using DNAMAN, Lasergene, Vector NTI and BLAST. The target sequence was selected and modified using plant preferred codons by the Synthetic Gene Designer and JCat (Java Codon Adaptation Tool) with the addition of a His-tag to the C-terminus to construct the full-length modified HPV18L1 (mHPV18L1). mHPV18L1 was divided into 5 large segments, namely LS1 to LS5, with sizes ranging from 204 to 477 bp. Forty-three small oligonucleotide fragments with sizes of 57-59 bp and 6 pairs of primers were designed and synthesized. mHPV18L1 was amplified by overlapping PCR and subcloned into pMD18-T vector. The recombinant plasmid was identified by restriction enzymes digestion and sequencing. RESULTS: mHPV18L1 was successfully assembled using overlapping PCR. The results of digestion with restriction enzymes and PCR amplification confirmed that the recombinant vector pMD18T- mHPV18L1 contained the inserts with expected size of 1749 bp. mHPV18L1 sequence was confirmed by sequencing. CONCLUSION: mHPV18L1 with plant preferred codons and the recombinant vector pMD18T- mHPV18L1 have been obtained. PMID- 19304508 TI - [Excretory/secretory antigens from Clonorchis sinensis induces hepatic fibrosis in rats]. AB - OBJECTIVE: To investigate the role of excretory/secretory antigens from Clonorchis sinensis (CsESAs) in hepatic fibrosis induced by C. sinensis infection in rats and explore the possible mechanism. METHODS: CsESAs was collected from adult C. sinensis cultured in sterile condition for 12 h and injected intraperitoneally in Wistar rats. Masson staining was used to observe the changes in the hepatic collagen fiber after the injection. HE staining and immunofluorescence staining were performed to detect the expression of alpha smooth muscle actin (alpha-SMA) to examine the proliferation and the activity of hepatic stellate cells. The specific antibody titer of CsESAs was determined using enzyme-linked immunosorbent assay to investigate the role of the antigen antibody complex in the development of hepatic fibrosis. RESULTS: After intraperitoneal injection of CsESAs, obvious hepatic fibrosis and hepatic stellate cell proliferation and activation were observed in the rat livers. The severity of the hepatic fibrosis was associated with the dose of CsESAs injected, whereas the titer of the specific antibody against CsESAs showed no direct relation to the hepatic fibrosis. CONCLUSION: Intraperitoneal injection of CsESAs can cause hepatic stellate cell activation and hepatic fibrosis in rats, but the antigen-antibody complex does not seem to play the key role in the activation of the hepatic stellate cells. PMID- 19304509 TI - [Preparation and bioactivity evaluation of streptavidin-tagged human interleukin 15 fusion protein]. AB - OBJECTIVE: To obtain streptavidin-tagged human interleukin-15 (SA/hIL15) fusion protein and evaluate its bioactivity. METHODS: pET24a-6His-SA-hIL-15 and pET32a hIL-15-SA-6His plasmids were constructed and expressed in BL 21(DE3) host bacteria to generate the fusion protein. The recombinant fusion protein IL-15/SA was purified using Ni-NTA affinity chromatography and refolded, and the efficiency of surface modification of the fusion protein on biotinylated cells was examined by fluorescence-activated cell sorting. CCK-8 method was used to evaluate the effect of IL-15/SA fusion protein in inducing the proliferation of human peripheral-blood lymphocyte (PBL) cells stimulated by PHA. RESULTS: The recombinant SA-hIL-15 and hIL15-SA fusion proteins were highly expressed in BL21(DE3) at about 20% of the total bacterial proteins. The purified hIL15-SA fusion protein exhibited a bifunctionality by promoting the proliferation of PBL cells activated by PHA and high-affinity binding to biotinylated cell surface mediated by SA, with a cell surface modification efficiency exceeding 95%. SA-hIL 15 showed a 4-fold higher hIL15 bioactivity than hIL15-SA. CONCLUSION: SA/hIL-15 bifunctional fusion protein has been successfully obtained to facilitate the future development of hIL-15-surface-modified cancer cell vaccine. PMID- 19304510 TI - [Effect of angiotensin II type 1 receptor and angiotensin-converting enzyme gene silencing on nuclear factor-kappaB activity in hepatic stellate cells]. AB - OBJECTIVE: To investigate the effect of angiotensin II (AngII) type 1 (AT-1) receptor and angiotensin-converting enzyme (ACE) gene silencing on nuclear factor kappaB (NF-kappaB) activity in hepatic stellate cells (HSCs). METHODS: pSilencer/AT-1 alpha receptor siRNA and pSilencer/ACE siRNA plasmids were transfected into cultured HSC-T6 cells, which were subsequently stimulated by 10( 6) mol/L AngII or ACE inhibitor (ACEI). The DNA binding activity of NF-kappaB in the transfected cells was analyzed using electrophoretic gel mobility shift assay (EMSA). RESULTS: s Gel shift studies showed that stimulation of the HSCs by AngII markedly increased the DNA-binding activity of NF-kappaB, which was inhibited by the transfection with pSilencer/ AT-1 alpha receptor siRNA plasmid or pSilencer/ACE siRNA plasmid. CONCLUSION: AT-1 alpha receptor and ACE gene silencing result in inhibition of NF-kappaB activity in HSCs in vitro. PMID- 19304511 TI - [p35 and p25 expressions and Cdk5 kinase activity in primary cultured rat hippocampal neurons with X-ray exposure]. AB - OBJECTIVE: To study the expressions of p35 and p25 and Cdk5 kinase activity in cultured rats hippocampal neurons following X-ray exposure to provide experimental evidence for prevention and treatment of radiation encephalopathy. METHODS: The hippocampal neurons cultured for 12 days were subjected to a single dose X-ray exposure of 30 Gy. Western blotting was used to detect the p35 and p25 protein levels, and the effect of pretreatment with roscovitine, a Cdk5 inhibitor, on the apoptosis of the hippocampal neurons following the exposure was examined with 4',6-diamidino-2-phenylindole (DAPI) staining. RESULTS: The protein level of p35 increased significantly 3.5 and 4 h after the irradiation by 1.51 /+0.13 and 1.45-/+0.14 folds in comparison with the control level, respectively (P<0.01), and the p25 level increased significantly 6 h after irradiation by 1.62 /+0.28 folds (P<0.05). Nuclear condensation occurred in (24.8-/+3.97)% of the neurons 24 h after 30 Gy X-ray exposure, a rate significantly higher than that in the nonexposed cells [(1.82-/+1.08)%, P<0.01) and that in roscovitine-pretreated neurons [(7.74-/+2.27)%, P<0.01). CONCLUSION: X-ray exposure activates Cdk5 by increasing the p35 and p25 expressions in rat hippocampal neurons, and inhibition of Cdk5 activity with roscovitine can significantly protect the neurons from apoptosis. PMID- 19304512 TI - [Construction of a eukaryotic expression vector for alpha-1-antitrypsin and the localization of the expression product in NIH 3T3 cells]. AB - OBJECTIVE: To construct a eukaryotic expression vector for alpha-1-antitrypsin (AAT) and detect its expression and localization in NIH 3T3 cells. METHODS: The total RNA was extracted from the liver tissue of BALB/c mice, and the corresponding coding sequences for mouse AAT (GenBank accession No. NM_009243) were amplified by RT-PCR and cloned into hemagglutinin (HA)-tagged vector pcDNA3 HA. The construct was then transfected into NIH 3T3 cells, which were observed under fluorescence microscope. RESULTS: The recombinant plasmid was verified by PCR, enzyme digestion and sequence analysis, and the fusion protein was highly expressed in NIH 3T3 cells. Under fluorescence microscope, the fusion protein was found to distribute mainly in the cytoplasm. CONCLUSION: The expression vector for AAT-HA fusion protein has been successfully constructed and effectively expressed in mammalian cells to allow future functional study of AAT in mammalian cells. PMID- 19304513 TI - [Expression, purification and refolding of streptavidin-tagged human tumor necrosis factor-alpha fusion protein]. AB - OBJECTIVE: To study the purification, refolding and bioactivity of streptavidin tagged human tumor necrosis factor-alpha (SA-TNF-alpha) bi-functional fusion protein. METHODS: SA-TNF-alpha fusion protein was expressed in BL21(DE3) host bacteria, purified using Ni-NTA affinity chromatography and refolded by dilution and dialysis followed by identification using Western blotting. The effect of SA TNF-alpha fusion protein against L929 cells was evaluated by MTT assay. Flow cytometry was used to analyze the surface modification of biotinylated MB49 tumor cells by SA-TNF-alpha fusion protein. RESULTS: Recombinant SA- TNF-alpha fusion protein was expressed in BL21(DE3) at about 30% of total bacterial protein, with a purity of about 95% after purification. The SA-TNF-alpha fusion protein existed as dimmers, tetramers and higher order structures after refolding. The fusion protein exhibited a bi-functionality by inhibiting L929 cells and SA-mediated high-affinity binding to biotinylated cell surfaces, with an anchor modification rate of above 90%. CONCLUSION: The dimmers, tetramers and higher order structures of the obtained SA-TNF-alpha fusion protein all exhibit a bi-functionality, and may serve as a potential candidate therapeutic agent for tumors. PMID- 19304514 TI - [PI3K p85alpha expression and its role in the progression of colorectal cancer]. AB - OBJECTIVE: To investigate the expression of PI3K p85alpha in normal colorectal tissue, colorectal adenoma and primary colorectal carcinoma and explore its significance in the progression of colorectal cancer. METHODS: The expression of PI3K p85alpha was detected in 116 normal colorectal tissue, colorectal adenoma and primary colorectal carcinoma specimens using immunohistochemical staining, and the relationship between the expression of PI3K p85alpha protein and the clinicopathological factors was analyzed. RESULTS: The positivity rates of the expression of PI3K p85alpha protein increased gradually in the progression of colorectal cancer and showed significant differences between the tissues (P<0.05). A significant difference was also noted in the positivity rates of the PI3K p85alpha expression in colorectal carcinoma tissues at different Dukes' stages (P<0.05). No obvious correlation was found between PI3K p85alpha expression and the degree of the tumor differentiation. CONCLUSIONS: Abnormal PI3K p85alpha expression occurs in the progression of colorectal cancer in close relation to the clinical stage, and the PI3K/AKT pathway plays an important role in the progression of colorectal cancer. PMID- 19304515 TI - [Isolation, culture and chondrogenic differentiation of goat bone marrow mesenchymal stem cells]. AB - OBJECTIVE: To explore the isolation, in vitro culture and chondrogenic differentiation of goat bone marrow mesenchymal stem cells (BMSCs). METHODS: Bone marrow was harvested from a 10-month-old Chinese goat for adherent culture of the BMSCs in vitro. Flow cytometry was performed to detect the cell surface markers of the BMSCs of the fourth generation. The induction medium (containing 10% fetal bovine serum, high-glucose DMEM, 6.25 microg/ml insulin, 6.25 microg/ml transferrin, 50 microg/ml vitamin C, 100 nmol/L DXM and 10 ng/ml transforming growth factor-beta1) was then applied for chondrogenic differentiation. Cytochemical staining, RT-PCR and Western blotting were performed to detect the expressions of type II collagen and aggrecan in the cells at the time points of 0, 1, 2 and 4 weeks. RESULTS: The goat BMSCs grow well in vitro with a high purity in the fourth generation. The expression of chondrocyte phenotypes were observed at 1, 2 and 4 weeks, which became more obvious as the culture prolonged. The mRNA and protein expression of type II collagen and aggrecan in the BMSCs increased obvious after the induction and had reached a satisfactory level by 2 weeks. CONCLUSION: Goat BMSCs have the potential to differentiate into chondrocytes in vitro, and the results of this study provide the experimental basis for application of goat BMSCs in bone and cartilage tissue engineering in vivo. PMID- 19304516 TI - [Effect of valproic acid on the expression of P27(Kip1) and P170 and drug resistance of HL-60/HT cells]. AB - OBJECTIVE: To investigate the effect of valproic acid on the expression of P27(Kip1) and P170 and drug resistance of leukemia HL60/HT cell line and explore its possible mechanisms. METHODS: HL-60/HT cells were derived from HL-60 cells induced by harringtonine (HT) in gradient concentrations. The inhibitory effect of valproic acid on the proliferation of HL-60 and HL-60/HT cells was evaluated by MTT assay, and the P27(Kip1) expression, P170 expression and cell cycle of the cells were analyzed with flow cytometry. RESULTS: The multidrug-resistant HL 60/HT was acquired, which showed a stable drug-resistant index with increased IC(50) of HT, VCR, DNR and Ara-c by 9.30, 5.20, 4.91 and 3.65 folds, respectively, as compared with those of HL60 cells. The expression of P27(Kip1) in HL-60/HT cells was significantly lower but P170 expression significantly higher than that of HL-60 cells and normal mononuclear cells (P<0.05). The expressions of P27(Kip1) and P170 showed no significant difference between normal mononuclear cells and HL-60 cells. The growth inhibition rate of VPA combined with Ara-C was significantly higher than that of valproic acid or Ara-C alone in HL-60/HT cells and HL-60 cells (q=1.37 and 1.51, respectively). HL-60/HT and HL 60 cells cultured in the presence of VPA resulted in a significant increase in the expression of P27(Kip1) and the G(1)-phase cells (P<0.05), but the expression of P170 underwent no significant changes (P>0.05). CONCLUSION: HL-60/HT cells have lower P27(Kip1) expression compared with HL-60 cells. Valproic acid can inhibit the growth of HL-60/HT cells and enhance their Ara-C sensitivity possibly by increasing P27(Kip1) expression and causing cell cycle arrest in G(1) phase. PMID- 19304517 TI - [Artemin and GFRalpha3 expressions and their relevance to perineural invasiveness and metastasis of pancreatic carcinoma]. AB - OBJECTIVE: To investigate the association of artemin and GFRalpha3 expressions with perineural invasion and metastasis of pancreatic carcinoma. METHODS: Semi quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression of artemin and GFRalpha3 in pancreatic carcinoma tissues, adjacent tissues and normal pancreas tissues, and the relevance of artemin and GFRalpha3 expressions to the perineural invasion and metastasis of pancreatic carcinoma were analyzed. RESULTS: The positivity rates of artemin and GFRalpha3 expressions were 72.09% and 67.44% in pancreatic carcinoma, respectively, significantly higher than those in the adjacent tissue (18.19% and 22.73%). The positivity rates of artemin and GFRalpha3 expressions were significantly higher in patients with perineural invasion than in those without perineural invasion (chi(2)=11.11 and 11.78, respectively, P<0.01). Significantly higher expression of artemin mRNA was noted in pancreatic carcinoma (0.741-/+0.014) than in the normal pancreas tissue (0.101-/+0.031, P<0.05), and patients with perineural invasion showed significantly higher positivity rates of artemin mRNA expression (0.843-/+0.012) than those without perineural invasion (0.512-/+0.017, P<0.05). CONCLUSION: Artemin and GFRalpha3 expressions may play an important role in perineural invasion of pancreatic carcinoma and can be used a useful indicators for evaluating the biological behavior of pancreatic carcinomas. PMID- 19304518 TI - [Construction of the eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 and green fluorescent protein]. AB - OBJECTIVE: To construct an eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and enhanced green fluorescent protein (EGFP) controlled by cytomegalovirus promoter using pIRES EGFP vector. METHODS: The mtHSP70 gene fragment was amplified by PCR from pVAX mtHSP70-HSV2gD using specific primers. The PCR product was cloned into the vector pMD 18-T vector, and the correct clone was selected according to DNA sequence analysis. The interested mtHSP70 gene fragment was subcloned into pCMV-IRES-EGFP vector with XhoI and EcoR I digestion. The recombinant plasmid was transfected into mouse melanoma B16 cell line, and the green fluorescent cells were detected by fluorescence microscopy and mtHSP70 expression was detected by Western blotting. RESULTS: The recombinant plasmid obtained was confirmed by enzyme digestion. The transfected mouse melanoma B16 cells exhibited green fluorescence under fluorescence microscopy and expressed mtHSP70 protein as demonstrated by Western blotting. CONCLUSION: The eukaryotic coexpression vector PCMV-mtHSP70 IRES-EGFP has been established to allow further investigation of the role of mtHSP70 gene in tumor immunotherapy. PMID- 19304519 TI - [Effect and safety of the therapies for acute myocardial infarction patients with failed thrombolytic therapy: a systematic review]. AB - OBJECTIVE: To assess the effect and safety of therapies in common use for acute myocardial infarction (AMI) patients with failed thrombolytic therapy. METHODS: We searched the Cochrane Central Register of Controlled Trials (CENTRAL, Issue 2, 2006), MEDLINE (1966 to July 2006), EMBASE (1984 to July 2006), China National Knowledge Infrastructure (CNKI, 1994 to July 2006), China Biomedicine Database disc (CBMdisc, 1980 to July 2006). We also searched several key Chinese journals in the field of cardiovascular diseases. The language was limited to Chinese and English. We included all the randomized controlled trials (RCTs) for acute myocardial infarction patients with failed thrombolytic therapy. Two authors independently assessed the methodological quality of the included studies, the data were analyzed by RevMan 4.2.8 from the Cochrane Collaboration. RESULTS: Nine RCTs met the inclusion criteria. A significant difference was found between the rescue percutaneous coronary intervention (PCI) group and conventional treatment group in the mortality rate at the end of the follow-up [RR=0.64, 95%CI (0.41, 0.98)]. Thromboembolic stroke and bleeding in rescue PCI group were significantly higher than that in conventional treatment group [RR=4.39, 95%CI (1.14, 16.87), and RR=2.79, 95%CI (1.55, 5.02), respectively]. Compared with conventional therapy, rescue thrombolytic treatment was associated with a significantly higher reperfusion rate [RR=2.92, 95%CI (1.75, 4.85)]. Comparison between rescue PCI with rescue thrombolytic treatment revealed that the revascularization rate in rescue PCI group was significantly lower than that in rescue thrombolytic group [RR=0.57, 95%CI (0.34, 0.95)], and the incidence of bleeding was significantly higher in rescue PCI group [RR=2.15, 95%CI (1.27, 3.63)]. Comparison of glycoprotein (GP)IIb/IIIa receptor antagonists with standard treatment showed no significant difference between them in the mortality rate and bleeding rate at the end of the follow-up. CONCLUSION: Current evidence does not confirm the effect or safety of the therapies for AMI patients with failed thrombolytic therapy, nor support the routine use of these therapies in clinical practice except for rescue PCI that reduces mortality compared with traditional treatment. Further high-quality randomized controlled trials are needed to provide reliable evidence for the treatments of AMI patients with failed thrombolytic therapy. PMID- 19304520 TI - [Time course of let-7a expression in the cell cycle of human cervical carcinoma HeLa cells]. AB - OBJECTIVE: To investigate the time course of let-7a microRNA expression in the cell cycle of HeLa cells. METHODS: HeLa cells were synchronized in G(1), S and G(2)/M phases using double-thymidine block, and the cell cycle phases were defined by flow cytometry. Real-time quantitative RT-PCR was used to examine the expression of let-7a in HeLa cells in different cell cycle phases. RESULTS: The synchronization rates of G(1), S and G(2)/M phases were 84.81%, 83.65% and 77.69%, respectively. Let-7a was constitutively expressed throughout the cell cycle in HeLa cells, but the expression levels in G(1) and S phases were lower than those in G(2)/M phase. CONCLUSIONS: Cell cycle can significantly influence the expression level of let-7a, which may provide new clues to the understanding of the cell cycle control mechanisms. PMID- 19304521 TI - [Construction of eukaryotic expression vectors of basic fibroblast growth factor and transfection in rabbit bone marrow stromal cells]. AB - OBJECTIVE: To investigate approach and possibility of transferring basic fibroblast growth factor (bFGF) gene into rabbit bone marrow stromal cells (BMSCs). METHODS: The eukaryotic expression vectors harboring bFGF cDNA were constructed and transfected into rabbit BMSCs mediated by liposome. The transcription and expression of bFGF gene in the transfected BMSCs were detected by means of morphological observation, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and RT-PCR. The changes in the biological characteristics of the transfected MSCs were also observed. RESULTS: Stable overexpression of bFGF protein was detected in the transfected BMSCs, which showed differentiation towards chondrocyte lineage. CONCLUSION: Stable expression of bFGF gene in transfected BMSCs can induce cell differentiation into chondrocyte lineage. PMID- 19304522 TI - [Magnetic resonance myocardial perfusion imaging for evaluating myocardial viability after myocardial infarction]. AB - OBJECTIVE: To assess the value of magnetic resonance (MR) myocardial perfusion imaging (MRMPI) in evaluating the myocardial viability in patients with myocardial infarction. METHODS: MRMPI was performed in 51 patients with myocardial infarction using a 1.5 T MR scanner. All the patients were examined using IR-turbo FLASH sequence during the first-pass and delayed phase 5-30 min after injection of 0.1 mmol/kg Gd-DTPA at the rate of 4 ml/s. The short axis images were acquired during the first-pass, and both the short axis and long axis images were obtained during the delayed phase. The left ventricular wall on the short-axis slice was divided into 8 segments. A correlative study of the results of the rest and stress (99m)Tc single photon emission computed tomography (SPECT) was carried out in 21 patients. RESULTS: In the 51 patients with myocardial infarction, 42(82.3%) showed hypoperfusion during the first-pass imaging and 50(98%) had delayed hyperenhancement. In the 21 patients receiving SPECT, 48 nonviable segments was detected among the 168 segments scanned by (99m)TcSPECT, and MRMPI showed delayed hyperenhancement in all the infracted areas. Of the 120 viable segments detected by rest and stress (99m)Tc SPECT, 97 segments (80.8%) were found to be free of delayed hyperenhancement by MRMPI. With the rest and stress (99m)Tc SPECT as the reference, the sensitivity and the specificity of MRMPI were 100.0% and 80.8%, respectively. CONCLUSION: MRMPI allows effective identification of the myocardial viability and nonviability as well as the severity and extent of the myocardial infraction. PMID- 19304523 TI - [Effect of resveratrol-induced FasL up-regulation on the apoptosis of pancreatic acinar cells in rats with severe acute pancreatitis]. AB - OBJECTIVE: To investigate the effect of resveratrol on the apoptosis of pancreatic acinar cells in rats with severe acute pancreatitis (SAP) and explore the mechanism of such effect. METHOD: SD rats with 3.5% sodium taurocholate induced SAP were treated with resveratrol, and the serum amylase was detected with automatic biochemistry analyzer. The apoptosis of the pancreatic acinar cells in the rats was detected by TUNEL assay, and the expression of Fas and FasL genes was determined by RT-PCR and Western blotting. The pathological changes of the pancreas were observed under optical microscope. RESULTS: Compared with SAP group, the resveratrol-treated rats showed obviously decreased serum amylase and scores for pancreatic histopathological lesions. Resveratrol treatment significantly increased the apoptotic indices of pancreatic acinar cells and the levels of FasL mRNA and protein in rats with SAP. CONCLUSION: Resveratrol produces important therapeutic effect on SAP in rats by inducing pancreatic acinar cell apoptosis possibly as a result of up-regulated FasL gene expression. PMID- 19304524 TI - [Immunomodulatory effects of Fomes fomentarius polysaccharides: an experimental study in mice]. AB - OBJECTIVE: To investigate the immunomodulatory effects of Fomes fomentarius polysaccharides (FFP) in mice. METHODS: MTT assay was employed to evaluate the in vitro metabolic activity of the mouse splenocytes treated with FFP at different concentrations, and the secretion of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and interleukin 2 (IL-2) from the cells were measured by enzyme-linked immunosorbent assay. The changes in the phagocytotic activity of mouse macrophage in response to FFP treatment were evaluated by phagocytosis percentage of chicken red blood cells (CRBCs). The effect of FFP on the humoral immunity was assessed in mice immunized with sheep red blood cells (SRBCs) by measuring the serum levels of specific antibody (hemolysin) against SRBCs. RESULTS: FFP at the concentrations of 25, 50, and 100 microg/ml all significantly enhanced the metabolic activity of mouse splenocytes in vitro and increased the production of TNF-alpha, IFN-gamma and IL-2. FFP treatment also markedly enhanced the metabolic activity of mouse peritoneal exudate cells and TNF-alpha production by the cells. At the doses of 25, 50, and 100 mg/kg, FFP significantly increased serum hemolysin level in mice immunized with SRBCs, and FFP at 50 and 100 mg/kg obviously increased the capacity of mouse peritoneal macrophages in vivo for CRBC phagocytosis. CONCLUSION: FFP can promote the secretion of TNF-alpha, IFN-gamma and IL-2 by mouse immunocytes and enhance mouse humoral immune response and the phagocytotic activity of the macrophages. PMID- 19304525 TI - [Isolation and characterization of human rheumatoid arthritis fibroblast-like synoviocytes]. AB - OBJECTIVE: To isolate and characterize human rheumatoid arthritis (RA) fibroblast like synoviocytes (FLSs). METHODS: The synovial membrane tissues were obtained from 4 RA patients, 1 chondroma patient and 1 healthy subject and FLS were isolated by means of tissue culture. The cell morphology was observed by phase contrast microscope and the cell surface markers were detected by flow cytometry. RESULTS: The FLSs were successfully cultured from the synovial membrane tissues with good cell homogeneity after the third passage. The FLSs of the 3rd to 7th passages were stable and proliferated actively, followed by slow proliferation and aging since the 8th passage. Flow cytometry showed that the 4th-passage FLSs from the RA patients contained 99.04% CD90(+) cells, 2.73% CD3(+) cells, 0.29% CD3(-)CD19(+) cells, 2.81% CD3(-)CD16(+)CD56(+) cells, 5.89% CD14(+) cells, and 54.17% CD55(+) cells. The presence of interleukin-1 receptor type I (IL-1RI, 158.63-/+20.32 pg/ml) and IL-1beta (4.67-/+0.82 pg/ml) were detected in the cell culture supernatant of the 4th-passage FLSs from the RA patients by enzyme-linked immunosorbent assay ELISA. CONCLUSION: FLSs from RA patients can be effectively culture by means of tissue culture, and the cultured FLSs show high expressions of CD90, IL-1RI and IL-1beta. PMID- 19304526 TI - [Effects of celecoxib on PGE2 synthesis and COX-2 and VEGF-C mRNA expression in Tca8113 cell lines]. AB - OBJECTIVE: To investigate the effect of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib on prostaglandin E(2) (PGE2) release and vascular endothelial growth factor C (VEGF-C) and COX-2 mRNA expression in Tca8113 cell lines. METHODS: MTT assay was used to analyze the proliferation of Tca8113 cells. The PGE2 level was detected with enzyme-linked immunosorbent assay (ELISA), and the expressions of COX-2 and VEGF-C mRNA were examined with RT-PCR. RESULTS: Celecoxib could induce inhibitory effects on the growth and PGE2 release in Tca8113 cells. The RT-PCR results showed that celecoxib significantly down regulated the expression of VEGF-C mRNA, but produced a weak effect on COX-2 mRNA expression. CONCLUSION: The inhibitory effect of celecoxib on Tca8113 cell growth and the expressions of VEGF-C and COX-2 may be related to the release of PGE2. PMID- 19304527 TI - [Estimation of the position of right-sided double-lumen endobronchial tubes with spirometry in elderly patients]. AB - OBJECTIVE: To study the accuracy of position estimation of right-sided double lumen endobronchial tubes (DLTs) without carinal hook in elderly patients according to the changes of inspiratory peak airway pressure (Ppeak), lung compliance (Cdyn), pressure-volume loop (P-V loop), and flow-volume loop (F-V loop). METHODS: Ninety-six elderly patients undergoing thoracic surgery were intubated with right-sided Mallinckrodt DLTs, the depth of which was determined based on the regressive equation. After 15 min of two lung ventilation (TLV), the Ppeak showed a more than 50% increase from the baseline in 22 patients when switching to one lung ventilation (OLV) with Cdyn less than 50% of the baseline and Ppeak in excess of 22 cmH(2)O. In these 22 cases, the position of the DLT was determined by fiberoptic bronchoscope (FOB) and adjusted to the precise position at 15 min of OLV. RESULTS: DLTs were inserted into the right middle bronchi in 19 cases and the right lower lobe bronchi in 3 cases. At 124-/+39 s after OLV, the SpO(2) began to decrease, and the Ppeak of OLV increased by 91.0% and Cdyn decreased by 62.7%. The slope of P-V curve was reduced and the P-V loop extended, with reduced increment of the expiratory limb and the area of the F-V loop, so that the tips of the DLT were withdrawn by 0.5 to 1.9 cm. The Ppeak of OLV increased only by 43.4% when DLT was in correct position, and was Cdyn decreased by 33.6% ( P<0.01). CONCLUSION: When changes of Ppeak and Cdyn in excess of above guidelines occurs after switching from TLV to OLV in the elderly patients, FOB should be applied to determine the tip position of DLT before SpO(2) reduction takes place. PMID- 19304528 TI - [Comparison of indirect immunofluorescence assay and ELISA for detecting antinuclear antibodies and anti-double-stranded DNA antibodies]. AB - OBJECTIVE: To compare indirect immunofluorescence assay (IIFA) and enzyme-linked immunosorbent assay (ELISA) for detecting antinuclear antibodies (ANA) and anti double-stranded DNA antibodies (anti-dsDNA). METHODS: A total of 125 serum samples were obtained from patients with established or suspected autoimmune disease, and 82 samples were used for ANA detection and 57 for anti-dsDNA detection using both IIFA and ELISA. Fourteen samples were examined for both ANA and anti-dsDNA. In cases where discrepancy occurred in the results by the two methods, extractable nuclear antigens were detected using immunoblotting. RESULTS: The positivity rate of ANA detected by IIFA and ELISA was significantly different (87.8% and 73.17%, respectively, P<0.01), but the positivity rate of anti-dsDNA was similar between IIFA and ELISA (77.19% and 71.93%, respectively, P>0.05). The percent agreement between the two testing methods with different cutoff values of ANA and anti-dsDNA showed significant differences (P<0.01), and for some uncommon patterns, the percent agreement of the two methods was lowered in ANA detection but remained unchanged for anti-dsDNA with different ANA patterns. High percent agreements of the two methods were obtained with the cutoff ANA titer of 1:100 and the cutoff anti-dsDNA value of weak positivity, but they demonstrated a significant difference in testing low-titer ANA and anti dsDNA. CONCLUSION: IIFA is more sensitive than ELISA in detecting the total ANA and anti-dsDNA. ELISA prescreening combined with IIFA can obtain the information of the nuclear pattern and allow the observation of the titer alterations. The combination of two or more testing methods can greatly enhance the accuracy of the results. PMID- 19304529 TI - [Creativity and personality analysis in 302 medical students using personality questionnaires]. AB - OBJECTIVE: To evaluate the creativity of medical students and study their personality characteristics using personality questionnaires. METHODS: This investigation was conducted among 310 medical students using 16 Personality Factor Questionnaire (16PF) and Eysenck Personality Questionnaire (EPQ). RESULTS: A total of 302 students completed the questionnaires. The male and female students showed significant differences in two sub-factors with similar creativity. Four subfactors were identified to positively or inversely correlate to the creativity. The number of male students without either high or low scores was greater than that of female students. The incidences of abnormal scores in 16PF showed significant difference between students with typical EPQ scores and those with atypical scores for introversion-extroversion. CONCLUSIONS: These medical students do not show high creativity in this study. Compared with the male medical students, the female students are more likely to have extroverted personality, and their scores for 16 basic personality factors easily exceed the normal ranges, suggesting the necessity of mental health education. The students with at least 5 abnormal scores in 16PF may show a typical introversion extroversion personality, and individual psychological counseling can be considered when necessary. PMID- 19304530 TI - [Isolation and identification of stem cells derived from human exfoliated deciduous teeth]. AB - OBJECTIVE: To isolate and identify stem cells from human exfoliated deciduous teeth (SHED). METHODS: Human pulp tissue were dissected and digested to obtain the single cell suspension. The cell morphology was observed and the clonality of the obtained cells was assessed. The phenotype of the cells was detected by immunohistochemistry and flow cytometry (FCM), and the cell cycle was analyzed. The in vitro differentiation of the cells into adipose tissue and formation of mineralization nodules were evaluated. RESULTS: Clonogenic assay showed the formation of 16-18 clones in every 10(3) plated cells derived from human exfoliated deciduous teeth. These cells were found to express the markers of mesenchymal stem cells with a multipotent differentiation potential. CONCLUSION: The cells isolated from human dental pulp are clonogenic and have multipotent differentiation potential, suggesting their identity of SHED. PMID- 19304531 TI - [Expression of ERK5 in multidrug-resistant hepatocellular carcinoma cell line]. AB - OBJECTIVE: To detect the expression of extracellular-signal regulated protein kinase 5 (ERK5) in multidrug-resistant hepatocellular carcinoma cell lines hepG2/ADM and BEL-7402/5-FU and in the parental cell lines (hepG2 and BEL-7402), and study the role of ERK5 in multidrug resistance of hepatocellular carcinoma. METHODS: MTT assay was used to determine the multidrug resistance (MDR) of hepG2/ADM and BEL-7402/5-FU cells. Real-time PCR and Western blotting were used to detect the expression of ERK5 at mRNA and protein levels in the 4 cell lines. RESULTS: The resistance indices to ADM, 5-FU, and CDDP was 12.34, 5.74, and 3.81 in hepG2/ADM cells, and the resistance indices to 5-FU, VCR, OHP, MTX, and ADM was 15.32, 10.08, 5. 85, 6.74 and 3.26 in BEL-7402 cells, respectively. Compared with those in hepG2 and BEL-7402 cells, the ERK5 gene was up-regulated at both mRNA and protein levels in HepG2/ADM cells, but down-regulated at the mRNA level and up-regulated at the protein level in BEL-7402/5-FU cells. CONCLUSIONS: ERK5 is related to multidrug resistance in HepG2/ADM and BEL-7402/5-FU cells, and may provide new clues for reversing multidrug resistance of hepatocellular carcinoma cells. PMID- 19304532 TI - [Effects of Ciwujia in inducing marrow stromal cell differentiation into neuron like cells in vitro]. AB - OBJECTIVE: To study the effect of the traditional Chinese herbal drug Ciwujia in inducing the differentiation of marrow stromal cells (MSCs) into neuron-like cells. METHODS: Rat MSCs isolated from the whole bone marrow were amplified by adherent culture in vitro and induced to differentiate into neuron-like cells using serum-free DMEM/F12 containing Ciwujia. The protein and mRNA expressions of nestin, beta-Tubulin III and glial fibrillary acidic protein (GFAP) in the differentiated cells were detected by indirect immunofluorescence method, Western blotting and RT-PCR. RESULTS: The third-passage MSCs showed positive expression rates for CD44 and CD54 beyond 90% with decreased CD14 expression rate to 2.37%. Induction by Ciwujia of the MSCs resulted in cell body shrinkage and protrusion of the cell processes resembling those of neurons. The differentiated cells were positive for nestin and beta-Tubulin III expression and negative for GFAP as shown by immunofluorescence assay, Western blotting and RT-PCR. CONCLUSION: Ciwujia can induce the differentiation of rat MSCs into neuron-like cells in vitro. PMID- 19304533 TI - [Clinical application of expansive unilateral open-door laminoplasty of the cervical spine with OsteoMed M3 plate and screws]. AB - OBJECTIVE: To introduce a new technique using OsteoMed M3 titanium plate and screws for fixation of the posterior elements in the open position after expansive unilateral open-door laminoplasty and evaluate its clinical effect. METHODS: Sixteen patients with multilevel cervical disc herniation and canal stenosis were treated with an expansive unilateral open-door laminoplasty using OsteoMed M3 plate and screws, and the follow-up period lasted for over 6 months. RESULTS: Most of the patients had marked neurological improvement after the surgery. The mean Japanese Orthopaedic Association (JOA) score of the patients increased significantly from 9.06-/+2.380 (range 5 to 13) before surgery to 13.63 /+1.408 (range 11 to 16) at the final follow-up (P<0.005), with a mean recovery rate of 57.5%. One patient without postoperative neurological improvement underwent an additional anterior multilevel corpectomy. Bone fusion of the surgical lamina was achieved in all the cases without canal stenosis. CONCLUSION: Unilateral open-door laminoplasty with OsteoMed M3 titanium plate and screw fixation can effectively maintain the expansion of the spinal canal and resist closure while preserving the spinal alignment and stability. This modified technique is easy to perform and is associated with a low complication rate. PMID- 19304534 TI - [Association of ATP-binding cassette transporter A1 R219K polymorphism with atrial fibrillation]. AB - OBJECTIVE: To investigate the association of ATP binding cassette transporter A1 (ABCA1) gene R219K polymorphisms with atrial fibrillation (AF) in Chinese population. METHODS: A total of 250 patients with AF and 250 control subjects were selected. Polymerase chain reaction-restricted fragments length polymorphism (PCR-RFLP) was used to determine the ABCA1 genotype, and the serum concentration of C-reactive protein (CRP) and high-density lipoprotein cholesterol (HDL-C) were measured in all the subjects. RESULTS: The frequency of the RR , RK , KK , allele R , allele K genotype of ABCA1 in AF group and control group was 42.0%, 42.8%, 15.2%, 34.0%, 43.2% and 22.8%, 63.4%, 36.6%, 55.6%, 44.4%, respectively. The frequency of the KK genotype was significantly higher in the control group than in the case group (P=0.03), and the frequency of the allele K genotype was significantly different between the two groups (P=0.012). The serum CRP concentrations was significantly higher in AF group than in the control group (P=0.004), but serum HDL-C level showed no difference between the two groups. The serum CRP concentrations were significantly higher in patients with RR genotype than in those with KK genotype (P=0.013), and patients with RR genotype had significantly lower HDL-C level than those with RK and KK genotypes (P=0.009 and 0.027, respectively). CONCLUSION: Patients with AF have elevated serum CRP level in comparison with healthy individuals, and the K allele of R219K polymorphism is an independent protective factor against AF. PMID- 19304535 TI - [Relationship between cellular immune response and apoptosis of the brain neurons after brain trauma in rats]. AB - OBJECTIVE: To investigate the relation between CD4(+) and CD8(+) T lymphocyte infiltration and apoptosis of the neurons in the local traumatic brain tissue after brain trauma in rats. METHODS: In rat models of brain trauma, the changes in the number of CD4(+) and CD8(+) T lymphocytes and the apoptosis of neurons in the local traumatic brain tissue were observed by immunohistochemistry at different time points after brain trauma. RESULTS: Twenty-four hours after brain trauma, a significant increase in the number of CD4(+) and CD8(+) T lymphocytes occurred in the injured brain tissue, both reaching the highest levels on day 10, at the point of which the number of CD4(+) cells increased by about 15 folds and that of CD8(+) cells by about 20 folds compared with the control groups. The CD4(+) and CD8(+) T lymphocytes both began to decrease 30 days after the injury. A similar pattern of alterations was found in the apoptosis of neurons in the local brain tissue. Correlation analysis demonstrated a close positive correlation between the changes in CD4(+) and CD8(+) lymphocyte numbers and the number apoptotic neurons in the injured brain tissue. CONCLUSIONS: Brain trauma induces obvious increases in CD4(+) and CD8(+) T lymphocytes and enhanced cellular immune response in the injured brain tissue to mediate neuronal apoptosis and further exacerbate the brain tissue injuries. PMID- 19304536 TI - [Comparison of the surgical approaches for nephrectomy in living related donors]. AB - OBJECTIVE: To compare the clinical effects and graft outcomes of 4 surgical approaches for nephrectomy in living related kidney donors. METHODS: Between June, 2004 and June, 2007, 119 living related kidney donors underwent nephrectomy via different surgical approaches, and their clinical data were retrospectively analyzed. Of these donors, 22 received retroperitoneal open nephrectomy, 21 had retroperitoneoscopic nephrectomy, 13 had hand-assisted laparoscopic nephrectomy, and 63 underwent transperitoneal open nephrectomy. The operating time, warm ischemia time of the graft, renal graft artery and vein lengths, reduction rate of recipient serum creatinine in the first 3 days after renal transplantation, mean hospital stay and complications of the donors were compared between the 4 surgical approaches. RESULTS: Open surgeries were associated with significantly shorter operating time (P=0.0033) and warm ischemia time of the graft (P=0.0001), longer hospital stay (P=0.0000), higher hospital expenses (P=0.0000), faster postoperative reduction of recipient serum creatinine (P=0.0001), and longer renal artery and vein lengths (P=0.0000 on the left and P=0.0001 on the right) than laparoscopic surgeries. In the laparoscopic surgery group, subcutaneous emphysema occurred in 1 case, DGF in 2 cases, and lumbar vein hemorrhage in 2 cases for which open surgery was performed. In the open surgery group, only one case required reoperation due to adrenal gland hemorrhage. All the kidney grafts were successfully harvested without other complications observed in the donors. CONCLUSIONS: Both open and laparoscopic surgeries are safe for nephrectomy in living related kidney donors, and the selection of the surgical approaches depends on the kidney and donor conditions and the surgical proficiency of the surgeons. PMID- 19304537 TI - [Laparoscopic placement of total peritoneum intraperitoneal onlay mesh in patients with inguinal hernia]. AB - OBJECTIVE: To study the efficacy of laparoscopic placement of total peritoneum intraperitoneal onlay mesh (TPIPOM) for treatment of inguinal hernia. METHODS: TPIPOM was placed laparoscopically in 125 cases of inguinal hernia, and the clinical outcomes of the patients were observed and compared with 64 patients receiving transabdominal preperitoneal laparoscopic mesh repair (TAPP) and 53 with total extraperitoneal laparoscopic hernioplasty (TEP). RESULTS: The laparoscopic operations were successfully performed in all the patients. In TPIPOM, TAPP and TEP groups, the operating time was 30.8-/+10.3 min, 68.4-/+22.4 min and 69.5-/+23.4 min (P<0.05), the mean hospital stay was 3.8-/+1.3 days, 4.3 /+1.5 days and 4.5-/+1.6 days (P<0.05), the average time to ambulation was 1.2 /+0.5 days, 1.8-/+0.7 days and 2.2-/+0.8 days (P<0.05), the duration of pain was 1.0-/+0.5 days, 1.6-/+0.9 days and 1.9-/+0.8 days (P<0.05), and the cost was 5000.8-/+800.5 yuan, 8000.5-/+950.6 yuan and 8900.2-/+750.3 yuan (P<0.05), respectively. No scrotum edema occurred in these patients. The patients were followed up for 59.9-/+6.5 months and recurrence was found. CONCLUSION: TPIPOM is safe and effective for management of inguinal hernia with such advantages as minimal invasion, simple procedures, shorter operation time, reduced relapse and quick recovery. PMID- 19304538 TI - [Sensitivity of Jatropha curcas seeds to (60)Co-gamma radiation and their medial lethal doses in radiation breeding]. AB - OBJECTIVE: To study the sensitivity of Jatropha curcas seeds from three different locations to (60)Co-gamma radiation and to determine the medial lethal doses (LD50) of (60)Co-gamma radiation for these seeds. METHODS: Six different radiation doses (0, 100, 150, 200, 250 and 300 Gy) were used. Based on the germination rate 50%, LD50 doses of (60)Co-gamma radiation for the seeds were calculated using linear regression equation. RESULTS: LD50 doses of (60)Co-gamma radiation for these seeds were 178 Gy (seeds from Guangdong), 132 Gy (seeds from Hainan) and 198 Gy (seeds from India) respectively. Increasing radiation doses caused more significant changes in leaf shape of the M1 seedlings. CONCLUSION: The results provides an important experimental basis for the radiation breeding of the important herbal and energy plant J. curcas. PMID- 19304539 TI - [Causal analysis of initial misdiagnosis of pulmonary embolism]. AB - OBJECTIVE: To analyze the causes of initial erroneous diagnosis of pulmonary embolism (PE) to improve the diagnostic efficiency. METHODS: The clinical data of 63 patients with a definite diagnosis of PE were retrospectively analyzed. According to the initial diagnosis, the patients were divided into definite diagnosis group (Group A, 23 cases) and misdiagnosis group (group B, 40 cases). The risk factors, initial symptoms, time of definite diagnosis, Wells scores, revised Geneva scores, and findings in chest X-ray and ECGs after onset and before the definite diagnosis were compared between the two groups. RESULTS: In group A, recent operations, malignancy, long-term bedridden state, PE history and deep vein thrombosis (DVT) symptom were more commonly seen than in group B, and the patients in group B were more likely to have hypertension, smoking, diabetes mellitus and lower limb varicose veins. The patients in group B had significantly lower Wells scores and revised Geneva scores than those in group A [2.50 (5.00) vs 6.00 (6.00), u=-3.296, P<0.001; 5.50 (4.75) vs 12.00 (9.00), u=-3.187, P<0.001, respectively]. In group B, chest examination in 22 of the 40 cases (55%) reported pulmonary infection, and among them, 15 were misdiagnosed as pneumonia. In groups A and B, SIQIIITIII/QIIITIII in ECG was found in 5 (21.7%) and 0 cases (0%), and normal ECG in 2 (8.7%) and 18 (45.0%) cases, respectively, showing significant difference between the two groups (P=0.010 and 0.003, respectively). CONCLUSION: The initial misdiagnosis of PE results mainly from the low awareness of some of the PE risk factors on the part of the physicians, atypical clinical manifestations and excessive dependence on chest films and ECGs. PMID- 19304540 TI - [Clinical characteristics and outcomes of 59 patients with acute lymphoblastic leukemia positive for BCR/ABL]. AB - OBJECTIVE: To study the clinical characteristics and outcomes of BCR/ABL-positive acute lymphoblastic leukemia (BCR/ABL360888725-ALL) and screen the prognostic factors for BCR/ABL360888725-ALL. METHODS: From January 2001 to May 2008, 59 patients (median age of 32 years ranging from 3 to 69 years) with the diagnosis of BCR/ABL360888725-ALL by fluorescence in situ hybridization received induction chemotherapy with VDLP-/+Ara-C regimen. The patients who failed to respond to the chemotherapy received subsequent consolidation chemotherapy with imatinib (400 800 mg/day) (17 cases) or allogeneic hematopoietic stem cell transplantation (allo-HSCT) (16 cases). RESULTS: Of the 59 patients, 32 (58.3%) achieved complete remission (CR) after the first induction cycle. In patients with peripheral white blood cell (WBC) count <30=10(9)/L, 30-99.9(9)/L and > or =100(9)/L, the CR rates were 75.0% (18/24), 56.3% (9/15) and 26.3% (5/19) (P=0.006), and the overall survival probability of 2 years ( OSs of 2-yrs) was 24.7%, 22.5% and 21.1%, respectively (P=0.180). According to the FAB classification, 56 cases were divided into L1, L2 and biphenotypic acute leukemia (BAL) subgroups, and their CR rates were 66.7% (6/9), 63.2% (24/38) and 22.2% (2/9) (P=0.029), with OSs of 2 yrs of 22.2%, 27.0% and 22.0%, respectively (P=0.623). In terms of immunophenotype grouping by EGIL, the patients with ALL, myeloid antigen-positive ALL and BAL had CR rates of 61.1% (11/18), 60.6% (20/33) and 12.5% (1/8) (P=0.039), and the OSs of 2-yrs of 22.7%, 21.0% and 18.8%, respectively (P=0.643). In 55 patients with known karyotype, the CR rates were 71.4%(5/7), 70.8% (17/24) and 37.5% (9/24) in normal, sole t(9;22) abnormality, t(9;22) with additional abnormalities groups (P=0.046), with the OSs of 2-yrs of 42.9%, 34.0% and 7.3%, respectively (P=0.000). The patients complicated by septicemia had significantly lower OSs of 2-yrs than those without septicemia (0% vs 38.8%, P=0.005). The OSs of 2-yrs were significantly higher in patients with consolidation chemotherapy with imatinib than those without (48.0% vs 11.2%, P=0.001), and allo-HSCT was associated with significantly higher OSs of 2-yrs than exclusive chemotherapy (54.2% and 8.5%, P=0.000). CONCLUSION: BCR/ABL360888725-ALL with WBC> or =100 x 10(9)/L, presence of BAL diagnosed by FAB or FACM, t(9;22) with additional chromosome abnormalities all adversely affect the treatment results, and additional chromosome abnormalities and septicemia are associated with lower OSs of 2-yrs. Imatinib treatment and allo HSCT can both improve the OSs of 2-yrs of the patients with BCR/ABL(+)-ALL. PMID- 19304541 TI - [Correlation of heart rate turbulence to coronary lesions and its changes following percutaneous transluminal coronary angioplasty]. AB - OBJECTIVE: To investigate the correlation between heart rate turbulence (HRT) and coronary lesion and the effects of percutaneous transluminal coronary angioplasty (PTCA) on HRT. METHODS: This study involved 150 patients undergoing 24-hour ambulatory electrocardiography (AECG) and elective coronary angiography (CAG). AECG was monitored on the first day and 7 days after PTCA in 108 patients with positive CAG findings, and the variation of HRT and cardiac functions were observed. The turbulence onset (TO), turbulence slope (TS) and turbulence timing (TT) of each section of HRT were calculated, analyzed and compared. RESULTS: The values of TO and TT were significantly higher and TS significantly lower in CAG positive group than in CAG-negative group (P<0.05 or 0.001). Significant difference was found in TO, TS and TT between patients with single and multiple coronary lesions (P<0.05 and 0.001). The values of TO, TS and TT on the first day after PTCA improved significantly in comparison with the those before PTCA in patients with single and multiple coronary lesions (P<0.001). Postoperative follow-up of the patients revealed obviously attenuated HRT in patients with left cardiac insufficiency compared with the patients with normal cardiac function (P<0.05). CONCLUSION: HRT is correlated to the severity of the coronary lesions and shows significant improvement after PTCA. Cardiac function insufficiency is an important factor affecting the HRT attenuation. PMID- 19304542 TI - [Expression of transient receptor potential subfamily mRNAs in rat testes]. AB - OBJECTIVE: To study the expression of the mRNAs of transient receptor potential (TRP) gene subfamily TRPV and TRPM in rat testes. METHODS: Normal SD rat testes were collected and the expression of TRPV and TRPM mRNAs were detected by routine RT-PCR. RESULTS: The TRPV4, TRPV5, TRPV6, TRPM3, TRPM4 and TRPM8 mRNAs were detected in the rat testes, but the other members of TRPV and TRPM family were not detected. CONCLUSIONS: TRPV4, TRPV5, TRPV6, TRPM3, TRPM4 and TRPM8 are expressed in rat testes. This finding provides the basis for exploring the functions of TRPV and TRPM in the testes and the relation between testis diseases and the TRP family. PMID- 19304543 TI - [Value of C-reactive protein in the diagnosis of arteriosclerosis in patients with essential hypertension]. AB - OBJECTIVE: To evaluate the value of C-reactive protein (CRP) in the diagnosis of arteriosclerosis in patients with essential hypertension. METHODS: This study included 771 patients with essential hypertension and 243 healthy individuals, and all the subjects were measured for carotid-femoral pulse wave velocity (cfPWV) and serum CRP level using enzyme-linked immunosorbent assay (ELISA). The value of CRP in the diagnosis of arteriosclerosis in patients with essential hypertension was evaluated according to the receiver operating characteristics (ROC) curve, and the diagnostic sensitivity and specificity was evaluated with cfPWV> or =9 m/s as the golden diagnostic standard. RESULTS: The hypertensive patients had significantly higher cfPWV and serum CRP concentration than the healthy individuals (16.51-/+1.6 vs 9.81-/+1.1, P<0.001; 4.96-/+1.15 vs 3.52 /+0.33, P<0.001, respectively). CRP showed significant positive correlations to systolic blood pressure (SBP) and pulse pressure (PP) (r=0.584, P<0.001; r=0.624, P<0.001), and when controlled for age, SBP and PP, CRP was found in close correlation to cfPWV (r=0.746, P<0.001). The AUCROC of CRP was 0.907, and the peak point of the ROC curve was 3.85 mg/L, at which point CRP showed a diagnostic sensitivity of 83.9% and specificity of 86.8% with a misdiagnosis rate of 13.2% for arteriosclerosis. CONCLUSIONS: Arteriosclerosis and nonspecific inflammation are prevalent in patients with essential hypertension, and CRP with the cutoff value of 3.85 mg/L may serve as a sensitive indicator for arteriosclerosis diagnosis in these patients. PMID- 19304544 TI - [Effects of Poly I:C in inducing growth inhibition and apoptosis of human hepatocellular carcinoma cells]. AB - OBJECTIVE: To explore the effect and mechanism of Poly I:C in inducing growth inhibition and apoptosis of human hepatocellular carcinoma SMMC-7721 cells. METHODS: SMMC-7721 cells were treated with different doses of Poly I:C for 24, 48, and 72 h, and the cell growth inhibition rate was analyzed with CCK-8 assay. The cell cycle and the apoptosis were analyzed using flow cytometry with Annexin V and PI staining, and quantitative RT-PCR analysis were used to detect the expression of TLR3, TRIF, and IFN-beta mRNA in cells. RESULTS: In the cells exposed to Poly I:C at low, moderate, and high doses, the inhibitory rates was the highest in high-dose Poly I:C group, and at a given Poly I:C dose, prolonged exposure resulted in significantly increased cell growth inhibition rate (P<0.05). Flow cytometry showed that Poly I:C induced cell apoptosis in a time- and dose-dependent manner and significantly increased the percentage of G1-phase cells as compared with that in the control group. The mRNA level of TLR3, TRIF, and IFN-beta were also increased following Poly I:C treatment in comparison with the control group. CONCLUSION: Poly I:C can induce significant growth inhibition and apoptosis of SMMC-7721 cells in a dose- and time-dependent manner possibly by causing cell cycle arrest and TLR3 signaling pathway activation that leads to IFN beta production and cell apoptosis. PMID- 19304545 TI - [Role of sodium cromoglycate in brain protection and its effects on tumor necrosis factor-alpha and interleukin-1beta after global cerebral ischemia reperfusion injury in gerbils]. AB - OBJECTIVE: To investigate the role of sodium cromoglycate in brain protection and its effects on brain tumor necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta) expressions after global cerebral ischemia-reperfusion (IR) injury in gerbils. METHODS: Twenty-four healthy male gerbils were randomized into 3 equal groups, namely the sham-operated group with isolation of the bilateral carotid arteries but without occlusion, IR injury model group with bilateral carotid artery occlusion, and sodium cromoglycate treatment group with bilateral carotid artery occlusion and sodium cromoglycate administration at 25 mg/kg via the lingual vein as soon as the reperfusion start with another dose 1 h later. The animals were then sacrificed and the thalamus were removed, fixed in 10% formaldehyde and sliced for observation under light microscope with HE staining. The rest brain tissues were prepared into homogenate to determine the content of TNF-alpha and IL-1beta. The right hemispheres of the gerbils were measured for wet weight and dry weight to calculate the water content in the brain. RESULTS: The water content in the brain of the gerbils in the model group was the highest among the groups, and that in sodium cromoglycate treatment group was significantly less than that of the model group (P<0.05). Microscopic examination showed the most severe brain tissue damage in the model group with also the highest TNF-alpha and IL-1beta levels in the brain. The brain TNF-alpha and IL 1beta levels in sodium cromoglycate group were significantly decreased as compared with those in the model group (P<0.05). CONCLUSION: Sodium cromoglycate can alleviate brain IR injury possibly by lowering the TNF-alpha and IL-1beta levels in the brain tissues. PMID- 19304546 TI - [Effects of losartan on left ventricular hypertrophy and plasma transforming growth factor-beta1 in elderly patients with hypertension]. AB - OBJECTIVE: To investigate the effects of losartan on left ventricular hypertrophy (LVH) and plasma transforming growth factor-beta1 (TGF-beta1) in elderly patients with essential hypertension (EH). METHODS: The elderly patients with EH were divided into two groups, namely EH+LVH group and EH group according to the data of echocardiogram. The systolic and diastolic blood pressures of the patients were monitored. Plasma TGF-beta1 was measured before and after 6 months' treatment with losartan, and the relationship between TGF-beta1 and other index were analyzed. RESULTS: After 6 months' treatment, the blood pressure of EH+LVH group and EH group were significantly lowered (P<0.01). Significant improvement of IVSTd, LVPWd, E/A, and LVMI (P<0.01) and obvious reduction of plasma TGF-beta1 (P<0.01) occurred in EH+LVH group after 6 months' treatment. Correlation analyses indicated that the plasma TGF-beta1 level was positively correlated to LVMI (P<0.01). CONCLUSION: Losartan can reversed LVH in elderly patients with EH partially by lowering plasma TGF-beta1 level. PMID- 19304547 TI - [Expression of special AT-rich sequence-binding protein mRNA and its clinicopathological significance in non-small cell lung cancer]. AB - OBJECTIVE: To detect the expression of AT-rich sequence-binding protein (SATB1) mRNA in non-small cell lung cancer (NSCLC) and explore the role of SATB1 in the development of NSCLC. METHODS: The total RNA was extracted from NSCLC tissues and normal lung tissues and reverse transcribed into cDNA. Real-time fluorescence quantitative RT-PCR was performed for detecting the expression of SATB1 mRNA these tissues. RESULTS: The expression of SATB1 mRNA was 13-fold higher in NSCLC tissues than in normal lung tissues (P<0.001), and in metastatic and nonmetastatic NSCLC, the expression was 23.63 and 5.57 folds that in normal lung tissues, respectively. CONCLUSION: SATB1 mRNA expression might be associated with the development and lymph node metastasis of NSCLC and may potentially used as an indicator for predicting the prognosis of NSCLC. PMID- 19304548 TI - [Effect of targeted argon-helium cryoablation on the portal region in canine livers]. AB - OBJECTIVE: To observe the effect of targeted argon-helium cryoablation on portal region of the liver in dogs by observing the pathological changes in the first order branches of the Glisson ductal system. METHODS: Twelve healthy dogs underwent percutaneous targeted argon-helium cryoablation of the liver and sacrificed at 3 and 28 days after the cryoablation to observe the pathological changes in target area for cryoablation and the first-order branches of the Glisson ductal system. RESULTS: No obvious damage was not found in the vascular wall of the portal vein by gross or microscopic observation, but the liver tissue in the vicinity of the blood vessels showed total necrosis. In spite of the injuries of different degrees in the first-order bile duct system after argon helium cryoablation, no severe damages such as perforation or full-thickness necrosis occurred in bile duct wall, and most of the injuries were temporary and reversible. The size of the ablated area on day 28 was significantly reduced as compared with that on day 3 following the cryoablation (P<0.05). In the acute stage after the cryoablation (1-3 days), ALT and AST levels increased significantly in (P<0.05) but recovered 1-4 weeks later (P>0.05). The cryoablated area was basically consistent with the pathological area that underwent necrosis (P>0.05). CONCLUSION: Targeted argon-helium cryoablation can cause total destruction of the liver tissue around the blood vessel without damaging the vascular walls of the portal vein. Argon-helium cryoablation induces relatively minor injuries to the bile duct of hepatic portal section and does not obviously damage the liver function, and the scope of tissue necrosis can be estimated according to the size of frozen area observed. Argon-helium cryoablation is a safe and minimally invasive operation with reliable therapeutic effect. PMID- 19304549 TI - [Expression and significance of interferon-inducible transmembrane protein-1 gene in Peutz-Jeghers syndrome]. AB - OBJECTIVE: To detect the mRNA and protein expression of interferon-inducible transmembrane protein-1 (IFITM1) in Peutz-Jeghers syndrome (PJS) and investigate the role of IFITM1 in the occurrence, development and carcinogenesis of PJS polyps. METHODS: Reverse transcription-PCR was employed to detect the mRNA expression of IFITM1 in 16 PJS polyp samples, adenomatous polyp tissues, colon adenocarcinoma samples, and normal intestinal mucosal tissues. The protein expression and localization of IFITM1 in these tissues (32 cases for each) were detected with immunohistochemical (IHC) staining. RESULTS: The IFITM1 mRNA expression was detected in all these tissues, and the expression intensity increased in the order of normal intestinal mucosa, PJS polyp, adenomatous polyp, and colon adenocarcinoma (F=92.704, P=0.000). IHC revealed that IFITM1 protein was localized mainly on the cell membrane and in the cytoplasm, with increased expression intensity in the same order as its mRNA and showing significant differences between the tissues by several rank-sum test (Kruskal-Wallis H, chi(2)=37.036, p=0.000). CONCLUSION: The expression level of IFITM1 is associated with the progression of the carcinogenetic process in PJS polyp, and can be used as a sensitive biomarker for diagnosis and prognostic evaluation of PJS. PMID- 19304550 TI - [Analysis of the factors contributing to diabetes insipidus after surgeries for craniopharyngiomas]. AB - OBJECTIVE: To analyze the factors contributing to the occurrence of diabetes insipidus after operations for craniopharyngiomas. METHODS: A total of 121 cases of diabetes insipidus following surgeries for craniopharyngiomas were retrospectively analyzed and the factors associated with postoperative diabetes insipidus were analyzed. RESULTS: The incidence of diabetes insipidus was 27.3% (33/121 cases) before the operation, 89.9% (107/1119) early after the operation and 39.8%(37/93) in later stages after the operation. The occurrence of early postoperative diabetes insipidus showed a significant relation to the classification and calcification of the craniopharyngioma. Patients with supradiaphragmatic and extraventricular tumors had the lowest incidence of postoperative diabetes insipidus. Late postoperative diabetes insipidus was closely correlated to such factors as age, classification of craniopharyngioma, and intraoperative treatment of the pituitary stalk, but not to the scope of tumor resection or tumor calcification. Late diabetes insipidus was more frequent in children and patients with severed pituitary stalk. The incidence of late postoperative diabetes insipidus was significantly higher in patients with supradiaphragmatic and extra-intraventricular tumors than in those with tumors beneath the diaphragma sellae and extraventricular tumors. CONCLUSIONS: Postoperative diabetes insipidus following surgeries for craniopharyngiomas is closely related to the tumor classification, calcification and pituitary stalk protection. PMID- 19304551 TI - [Establishment and assessment of Caco-2 cell in vitro absorption model]. AB - OBJECTIVE: To establish and assess the Caco-2 cell in vitro absorption model. METHODS: Caco-2 cells were cultured on the millipore filters fixed in Snapwell transport chamber. The cell morphology, transepithelial electrical resistance, mannitol efflux rate and alkaline phosphatase activities were monitored during culture. RESULTS: After 21 days of in vitro culture, formation of tight junction was observed between the cells. The transepithelial electrical resistance reach a relatively stable value of 620-/+47 Omega.cm(2), the mannitol efflux rate was lower than 0.3%.h(-1).cm(-2), and the alkaline phosphatase activity in the apical side was significantly higher than that in the basolateral side. CONCLUSION: The established Caco-2 cell model shows similar morphology to intestinal epithelial cells with formation of polarity, and can be used as an in vitro model for absorption studies. PMID- 19304552 TI - [Determination of ferulic acid in Fuan oral liquid by high-performance capillary electrophoresis]. AB - OBJECTIVE: To establish a quality control method of Fuan oral liquid. METHODS: High-performance capillary electrophoresis (HPCE) was used to determine the content of feruild acid in Herba taraxaci, and thin-layer chromatography (TLC) was performed to identify Herba taraxaci and Bupleurum chinense. The condition of HPCE was optimized with a fuse silica capillary tube (70 microm x 60 cm) and 20 mmol/L sodium tetraborate buffer (pH9.18) at a constant voltage of 12 kV and temperature at 25 degrees celsius;, with the detection wavelength at 313 nm. RESULTS: Clear spots were displayed on TLC. The calibration curve was linear in the range of 8-40 microg/ml for ferulic acid (Y=360.5-207.4, r=0.9997, n=5). The average recovery rate exceeded 95% with a RSD<3% (n=3). CONCLUSION: This method is simple and specific with a good reproducibility for quality control of Fuan oral liquid. PMID- 19304553 TI - [Audiological assessment in 88 patients with otosclerosis]. AB - OBJECTIVE: To analyze the clinical data of the hearing in otosclerosis patients and explore their association with the clinical manifestations. METHOD: A retrospective analysis of 88 otosclerosis cases (162 ears) was performed. Pure tone audiometry was performed in all the cases 1 to 3 days before the operation for speech analysis at the frequencies of 500, 1 k, 2 kHz and to determined the average pure-tone hearing threshold (PTA) and gas Bone gap (ABG). The incidence of carhart notch was observed in patients with simple conductive hearing loss and mixed hearing loss, and in the early, middle and late stage of otosclerosis. RESULTS: Carhart notch occurred at significant higher incidence in patients with simple conductive hearing loss and in early otosclerosis (P<0.05). CONCLUSION: For common otosclerosis, the lesion invades initially the vestibular window, round ligament and stapes floor, followed by the base of the cochlea, and therefore observation of the carhart notch may help determine the disease course. PMID- 19304554 TI - Formulation development of oral controlled release tablets of hydralazine: optimization of drug release and bioadhesive characteristics. AB - The current study involves development of oral bioadhesive hydrophilic matrices of hydralazine hydrochloride, and optimization of their in vitro drug release profile and ex vivo bioadhesion against porcine gastric mucosa. A 32 central composite design was employed to systematically optimize the drug delivery formulations containing two polymers, viz., carbomer and hydroxypropyl methyl cellulose. Response surface plots were drawn and optimum formulations were selected by brute force searches. Validation of the formulation optimization study indicated a very high degree of prognostic ability. The study successfully undertook the development of an optimized once-a-day formulation of hydralazine with excellent bioadhesive and controlled release characteristics. PMID- 19304555 TI - Development and biopharmaceutical evaluation of extended release formulation of tramadol hydrochloride based on osmotic technology. AB - Extended release formulation of tramadol hydrochloride (TRH) based on osmotic technology was developed and evaluated. Target release profile was selected and different variables were optimized to achieve it. Formulation variables such as the level of swellable polymer, plasticizer and the coat thickness of semipermeable membrane (SPM) were found to markedly affect drug release. TRH release was directly proportional to the levels of plasticizer but inversely proportional to the levels of swellable polymer and coat thickness of SPM. Drug release from developed formulations was independent of pH and agitation intensity but dependent on osmotic pressure of the release media. In vivo study was also performed on six healthy human volunteers and various pharmacokinetic parameters (cmax, tmax, AUC0-24, MRT) and relative bioavailability were calculated. The in vitro and in vivo results were compared with the performance of two commercial TRH tablets. The developed formulation provided more prolonged and controlled TRH release compared to the marketed formulation. In vitro-in vivo correlation (IVIVC) was analyzed according to the Wagner-Nelson method. The optimized formulation (batch IVB) exhibited good IVIV correlation (R = 0.9750). The manufacturing procedure was found to be reproducible and formulations were stable over 6 months of accelerated stability testing. PMID- 19304556 TI - Investigation of the structural requirement for inhibiting HIV integrase: QSAR study. AB - HIV integrase has emerged as a promising target for discovery of agents against the acquired immunodeficiency syndrome (AIDS) pandemic. With the purpose of designing new chemotypes with enhanced potencies against the HIV integrase enzyme, the QSAR study carried out on 37 novel phthalimide derivatives is presented. The developed QSAR model was validated by standard statistical parameters and through a detailed structural study of how it reproduces and explains the quantitative differences seen in experimentally known pharmacological data. The model showed a good correlative and predictive ability having a cross-validated correlation coefficient (r2 cv) of 0.709 and a conventional correlation coefficient (r2) of 0.949. The predictive correlation coefficient (r2 pred) was found to be 0.512. The study revealed that the antiretroviral activity is predominantly explained by the substituent size, shape and polarity and provided insights into how modulation of the steric bulkiness and polarities of the substituents could be made to optimize the integrase inhibitor interaction chemistry. A detailed investigation was made of the structural basis for the antiretroviral activity and the findings from the study could be usefully employed to design antagonists with a much more enhanced potency and selectivity. PMID- 19304557 TI - Preclinical studies of [61Cu]ATSM as a PET radiopharmaceutical for fibrosarcoma imaging. AB - [61Cu]diacetyl-bis(N4-methylthiosemicarbazone) ([61Cu] ATSM) was prepared using in house-made diacetyl-bis(N4-methylthiosemicarbazone) (ATSM) ligand and [61Cu]CuCl2 produced via the natZn(p, x)61Cu (180 muA proton irradiation, 22 MeV, 3.2 h) and purified by a ion chromatography method. [61Cu]ATSM radiochemical purity was >98 %, as shown by HPLC and RTLC methods. [61Cu]ATSM was administered into normal and tumor bearing rodents for up to 210 minutes, followed by biodistribution and co-incidence imaging studies. Significant tumor/non-tumor accumulation was observed either by animal sacrification or imaging. [61Cu]ATSM is a positron emission tomography (PET) radiotracer for tumor hypoxia imaging. PMID- 19304558 TI - Preparation and characterization of gliclazide-polyethylene glycol 4000 solid dispersions. AB - The objective of the present investigation was to study the effect of polyethylene glycol 4000 (PEG 4000) on in vitro dissolution of gliclazide from solid dispersions. Initial studies were carried out using physical mixtures of the drug and carrier. Solid dispersions were prepared by the melting or fusion method.Phase and saturation solubility study, in vitro dissolution of pure drug, physical mixtures and solid dispersions were carried out. PEG was found to be effective in increasing the dissolution of gliclazide in solid dispersions when compared to pure drug. FT-IR spectroscopy, differential scanning calorimetry and X-ray diffractometry studies were carried out in order to characterize the drug in the physical mixtures and solid dispersions. Dissolution enhancement was attributed to decreased crystallinity of the drug and to the wetting and solubilizing effect of the carrier from the solid dispersions of gliclazide. In conclusion, dissolution of gliclazide can be enhanced by the use of hydrophilic carrier. PMID- 19304559 TI - Potential prophylactic role of aminoguanidine in diabetic retinopathy and nephropathy in experimental animals. AB - This study sought to determine the potential prophylactic and antioxidant effects of aminoguanidine in experimentally induced diabetes. Four groups of Wistar rats, each composed of ten rats, were used. Two groups served as control. In group 3, diabetes was induced by a single intraperitoneal injection of streptozotocin (65 mg kg-1). In group 4, diabetes was induced and treated with aminoguanidine (100 mg kg-1 daily) orally for 3 months. Levels of serum glucose, glutathione peroxidase, glutathione reductase and erythrocytes catalase were analyzed on day 90 of the experiment. Retinal and kidney specimens were examined histopathologically after sacrifice of the animals. A significant antioxidant effect of aminoguanidine and its prophylactic role in diabetic retinopathy and nephropathy were observed in experimental animals. PMID- 19304560 TI - Synthesis and pharmacological evaluation of 3-cyclohexyl-2-substituted hydrazino 3H-quinazolin-4-ones as analgesic and anti-inflammatory agents. AB - A series of novel 3-cyclohexyl-2-substituted hydrazino-quinazolin-4(3H)-ones were synthesized by reacting the amino group of 3-cyclohexyl-2-hydrazino quinazolin 4(3H)-one with a variety of aldehydes and ketones. The starting material, 3 cyclohexyl-2-hydrazino quinazolin-4(3H)-one, was synthesized from cyclohexyl amine. Title compounds were investigated for analgesic, anti-inflammatory and ulcerogenic behavior. The compound 3-cyclohexyl-2-(1-methylbutylidene-hydrazino) 3H-quinazolin-4-one (4c) emerged as the most active compound of the series and is moderately more potent in its analgesic and anti-inflammatory activities compared to the reference standard diclofenac sodium. Interestingly, test compounds showed only mild ulcerogenic potential when compared to acetylsalicylic acid. PMID- 19304561 TI - Hepatoprotective effects of Calotropis gigantea extract against carbon tetrachloride induced liver injury in rats. AB - Ethanolic extract (50 %) of stems of Calotropis gigantea R. Br. (Asclepiadaceae) at doses of 250 and 500 mg kg-1 were studied for hepatoprotective activity in male Wistar rats with liver damage induced using carbon tetrachloride, 2 mL kg-1 twice a week. The protective effect of C. gigantea extract was compared with the standard drug silymarin. Various biochemical parameters such as aspartate amino transferase (AST), alanine amino transferase (ALT), glutathione (GSH), lipid peroxide (LPO), superoxidedismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were evaluated. The results revealed that the C. gigantea extract significantly decreased AST, ALT (p < 0.001) and lipid peroxide (p < 0.01) levels. The antioxidant parameters GSH, GPx, SOD and catalase levels were increased considerably compared to their levels in groups not treated with C. gigantea extract. PMID- 19304562 TI - Synthesis and pharmacological investigation of novel 4-(3-ethylphenyl)-1 substituted-4H-[1,2,4]triazolo[4,3-a] quinazolin-5-ones as a new class of H1 antihistaminic agents. AB - A series of novel 4-(3-ethylphenyl)-1-substituted-4H-[1,2,4] triazolo[4,3 a]quinazolin-5-ones (4a-j) were synthesized by the cyclization of 3-(3 ethylphenyl)-2-hydrazino-3H-quinazolin-4-one (3) with various one-carbon donors. The starting material, compound 3, was synthesized from 3-ethyl aniline by a new innovative route with improved yield. When tested for their in vivo H1 antihistaminic activity on conscious guinea pigs, all test compounds protected the animals from histamine induced bronchospasm significantly. Compound 4-(3 ethylphenyl)-1-methyl-4H - [1,2,4]triazolo[4,3-a]quinazolin-5-one (4b) emerged as the most active compound of the series and it is more potent (74.6 % protection) compared to the reference standard chlorpheniramine maleate (71 % protection). Compound 4b shows negligible sedation (10 %) compared to chlorpheniramine maleate (30 %). Therefore compound 4b can serve as the leading compound for further development of a new class of H1-antihistamines. PMID- 19304563 TI - Macromolecular prodrugs. XII. Primaquine conjugates: synthesis and preliminary antimalarial evaluation. AB - New primaquine conjugates 5-7 with glucosamine and two polymers of polyaspartamide type, poly[alpha,beta-(N-2-hydroxyethyl-DL-aspartamide)] (PHEA) and poly[alpha,beta-(N-3-hydroxypropyl-DL-aspartamide)] (PHPA), were synthesized, characterized and screened for their antimalarial activity. The conjugates differed in the type of covalent bonding, length of the spacer between the polymeric carrier and drug, molecular mass and drug-loading. Blood-schizontocidal activity of the prepared conjugates was tested against Plasmodium berghei infection in Swiss mice. Polymeric conjugates showed better antimalarial activity than the glucosamine conjugate. PMID- 19304564 TI - An unusual case of progressive shock and highly elevated procalcitonin level. AB - A 21-year-old man with signs and symptoms of rapidly progressive shock was admitted to the intensive care unit for treatment of suspected sepsis. Levels of inflammatory markers (including procalcitonin) were highly elevated, but no obvious focus of infection was apparent. Initial sepsis therapy included administration of broad-spectrum antibiotics, vasoconstrictors, and drotrecogin alfa. Cultures of blood, sputum, and urine showed no growth, and no viruses were detected. The random (no stimulation with corticotropin) cortisol level at admission was less than 25 nmol/L. Assays for autoantibodies to the adrenal cortex were strongly positive and confirmed the diagnosis of adrenal failure caused by Addison disease. After initiation of steroid therapy, the patient fully recovered. Although increased procalcitonin levels are considered a reliable and specific indicator of severe generalized infections and bacterial sepsis, elevated procalcitonin levels cannot be relied on when trying to differentiate between addisonian crisis and septic shock. PMID- 19304565 TI - Effect of mechanical chest percussion on intracranial pressure: a pilot study. AB - BACKGROUND: Treatment of brain injury is often focused on minimizing intracranial pressure, which, when elevated, can lead to secondary brain injury. Chest percussion is a common practice used to treat and prevent pneumonia. Conflicting and limited anecdotal evidence indicates that physical stimulation increases intracranial pressure and should be avoided in patients at risk of intracranial hypertension. OBJECTIVES: To explore the safety of performing chest percussion for patients at high risk for intracranial hypertension. METHODS: A total of 28 patients with at least 1 documented episode of intracranial hypertension who were having intracranial pressure monitored were studied in a prospective randomized control trial. Patients were randomly assigned to either the control group (no chest percussion) or the intervention group (10 minutes of chest percussion at noon). Intracranial pressure was recorded once a minute before, during, and after the intervention. RESULTS: Mean intracranial pressures for the control group before, during, and after the study period (14.4, 15.0, and 15.9 mm Hg, respectively) did not differ significantly from pressures in the intervention group (13.6, 13.7, and 14.2 mm Hg, respectively). CONCLUSIONS: Mechanical chest percussion may be a safe intervention for nurses to use on neurologically injured patients who are at risk for intracranial hypertension. PMID- 19304566 TI - Acute respiratory distress syndrome after zinc chloride inhalation: survival after extracorporeal life support and corticosteroid treatment. AB - No standard protocol exists for the treatment of acute respiratory distress syndrome induced by inhalation of smoke from a smoke bomb. In this case, a 23 year-old man was exposed to smoke from a smoke grenade for approximately 10 to 15 minutes without protective breathing apparatus. Acute respiratory distress syndrome developed subsequently, complicated by bilateral pneumothorax and pneumomediastinum 48 hours after inhalation. Despite mechanical ventilation and bilateral tube thoracostomy, the patient was severely hypoxemic 4 days after hospitalization. His condition improved upon treatment with high-dose corticosteroids, an additional 500-mg dose of methylprednisolone, and the initiation of extracorporeal life support. Arterial oxygenation decreased gradually after abrupt tapering of the corticosteroid dose and discontinuation of the life support. On day 16 of hospitalization, the patient experienced progressive deterioration of arterial oxygenation despite the intensive treatment. The initial treatment regimen (ie, corticosteroids and extracorporeal life support) was resumed, and the patient's arterial oxygenation improved. The patient survived. PMID- 19304567 TI - Single, intra-articular treatment with 6 ml hylan G-F 20 in patients with symptomatic primary osteoarthritis of the knee: a randomised, multicentre, double blind, placebo controlled trial. AB - OBJECTIVES: The primary objective was to compare a single, 6 ml, intra-articular injection of hylan G-F 20 with placebo in patients with symptomatic knee osteoarthritis. The safety of a repeat injection of hylan G-F 20 was also assessed. METHODS: Patients with primary osteoarthritis knee pain were randomly assigned to arthrocentesis plus a 6 ml intra-articular injection of either hylan G-F 20 or placebo in a prospective, double-blind (one injector/one blinded observer) study. RESULTS: were evaluated at 4, 8, 12, 18 and 26 weeks post injection. The primary outcome criterion was change from baseline over 26 weeks in Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index A pain. Secondary outcome measures included WOMAC A1 and C, patient global assessment (PGA) and clinical observer global assessment (COGA) and Outcome Measures in Rheumatology, Osteoarthritis Research Society International responder rates. A 4 week, open, repeat treatment phase evaluated safety only. Results: A total of 253 patients (Kellgren-Lawrence grade II or III) was randomly assigned. Patients receiving hylan G-F 20 experienced statistically significantly greater improvements in WOMAC A pain scores (-0.15, SE 0.076, p = 0.047), and several of the secondary outcome measures (WOMAC A1, PGA and COGA), than patients receiving placebo. There was no difference between the safety results of the two groups. No increased risk of local adverse events was observed in the open, repeat treatment phase. CONCLUSIONS: This placebo-controlled study demonstrated that, in patients with knee osteoarthritis, a single 6 ml intra-articular injection of hylan G-F 20 is safe and effective in providing statistically significant, clinically relevant pain relief over 26 weeks, with a modest difference versus placebo. PMID- 19304568 TI - Blockade of Dickkopf (DKK)-1 induces fusion of sacroiliac joints. AB - OBJECTIVE: To study whether Dickkopf (DKK)-1, an inhibitor of wingless (Wnt) signalling, is involved in the fusion of sacroiliac joints. METHODS: Mice transgenic for tumour necrosis factor (TNFtg mice), which develop bilateral sacroiliitis, were treated with vehicle, anti-TNF antibody or anti-DKK1 antibody. Sacroiliac joints were analysed for histological signs of inflammation, bone erosion, osteoclast formation and ankylosis. Moreover, expression of collagen type X, beta-catenin and DKK-1 was assessed by immunohistochemistry. RESULTS: There were no signs of spontaneous ankylosis of the sacroiliac joints in TNFtg mice. TNF blockade effectively reduced inflammation, bone erosion and osteoclast numbers in the sacroiliac joints, but did not lead to ankylosis. Blockade of DKK1 had no effect on inflammatory signs of sacroiliitis, but significantly reduced bone erosions and osteoclast counts. Moreover, DKK1 blockade promoted expression of collagen type X, the formation of hypertrophic chondrocytes and ankylosis of sacroiliac joints. CONCLUSION: DKK1 influences inflammatory remodelling of sacroiliac joints by prevention of joint ankylosis. This may indicate an important role of the Wnt signalling pathway in the structural bone changes of axial joint disease. Although this model does not reflect the entire spectrum of ankylosing spondylitis in humans, it helps to explain the pathophysiological processes of sacroiliac joint ankylosis, which is a hallmark of the spondyloarthritides. PMID- 19304569 TI - Mutations in the urocanase gene UROC1 are associated with urocanic aciduria. AB - Urocanase is an enzyme in the histidine pathway encoded by the UROC1 gene. This report describes the first putative mutations, p.L70P and p.R450C, in the coding region of the UROC1 gene in a girl with urocanic aciduria presenting with mental retardation and intermittent ataxia. Computed (in silico) predictions, protein expression studies and enzyme activity assays suggest that none of the mutations can produce a fully functional enzyme. The p.L70P substitution, which probably implies the disruption of an alpha-helix in the N-terminus, would alter its properties and therefore, its function. The p.R450C change would render impossible any interaction between urocanase and its substrate and would loss its enzyme activity. Consequently, these studies suggest that both mutations could alter the correct activity of urocanase, which would explain the clinical and biochemical findings described in this patient. PMID- 19304570 TI - Augmented blood pressure response to exercise is associated with improved long term survival in older people. AB - OBJECTIVE: Studies on the prognostic importance of the systolic blood pressure (SBP) response during exercise report ambiguous results. Most research focuses on younger and middle-aged selected patient groups and rarely includes women. We investigated the prognostic value of SBP response during exercise testing in 75 year-olds. DESIGN: Prospective observational cohort study. SETTING: A community based random sample of 75-year-old men and women (n = 382). MAIN OUTCOME MEASURES: The prognostic value of SBP change from rest to peak exercise during a symptom-limited cycle test was evaluated for the endpoints all-cause mortality and cardiovascular mortality during long-term follow-up. RESULTS: After a median follow-up of 10.6 years, 140 (37%) of the participants had died, 64 (17%) from cardiovascular causes. The all-cause mortalities for exercise SBP changes of < or =30 mm Hg, 31-55 mm Hg and >55 mm Hg were 5.1, 4.2 and 2.6 per 100 person-years, respectively (logrank 9.6; p = 0.008). For every 10 mm Hg increase in SBP during exercise the relative hazard for all-cause mortality was reduced by 13% (p = 0.030) and for cardiovascular mortality by 26% (p = 0.004) after adjustment for sex, smoking, waist circumference, total/HDL cholesterol ratio, prevalent ischaemic heart disease, hypertension, diabetes, cardiovascular medication, pre exercise SBP, exercise capacity, resting left ventricular ejection fraction and left ventricular mass index. CONCLUSIONS: Our findings suggest that an augmented SBP response during exercise is associated with an improved long-term survival among community-living 75-year-old individuals. PMID- 19304571 TI - Socio-economic status and early outcome from coronary artery bypass grafting. AB - OBJECTIVE: To determine the effects of socio-economic status (SES) on the outcome of coronary artery bypass grafting (CABG). DESIGN: Prospective cohort study. SETTING: Regional cardiac surgical unit. PATIENTS: 1994 consecutive patients undergoing non-emergency CABG. MEASURES: SES was determined from the patient's postcode using Carstairs tables. The primary end-point was all-cause mortality at 30 days. RESULTS: There were 50 deaths (2.5%) within 30 days of surgery. A higher Carstairs score demonstrated a trend towards increased 30-day mortality (odds ratio (OR) 1.09 per unit, 95% CI 1.00 to 1.20, p = 0.06). In a backward conditional model, including other predictors of early mortality, Carstairs scores were independently predictive (OR 1.12 per unit, 95% CI 1.01 to 1.24, p = 0.02). In a model including only Carstairs scores and the EuroSCORE, both were independent predictors of this outcome (OR for Carstairs score 1.11 per unit, 95% CI 1.00 to 1.22, p = 0.04). The 30-day mortality increases in each quartile of Carstairs scores, with patients in quartile 4 (most deprived) at significantly higher risk compared with quartile 1 (uncorrected OR 2.53 per unit, 95% CI 1.04 to 6.15; OR corrected for EuroSCORE, 2.56 per unit, 95% CI 1.03 to 6.34, p = 0.04 for both). Similarly, patients in the least affluent quartile were twice as likely to suffer a serious complication as those in the most affluent quartile (OR 2.14 per unit, 95% CI 1.32 to 3.46, p = 0.002). This increased risk was also independent of the EuroSCORE. CONCLUSIONS: Lower SES is associated with a poorer early outcome following CABG and is independent of other recognised risk factors. PMID- 19304572 TI - Socio-economic deprivation and outcome in patients with coronary artery disease. PMID- 19304573 TI - Chylomicronemia with a mutant GPIHBP1 (Q115P) that cannot bind lipoprotein lipase. AB - OBJECTIVE: GPIHBP1 is an endothelial cell protein that binds lipoprotein lipase (LPL) and chylomicrons. Because GPIHBP1 deficiency causes chylomicronemia in mice, we sought to determine whether some cases of chylomicronemia in humans could be attributable to defective GPIHBP1 proteins. METHODS AND RESULTS: Patients with severe hypertriglyceridemia (n=60, with plasma triglycerides above the 95th percentile for age and gender) were screened for mutations in GPIHBP1. A homozygous GPIHBP1 mutation (c.344A>C) that changed a highly conserved glutamine at residue 115 to a proline (p.Q115P) was identified in a 33-year-old male with lifelong chylomicronemia. The patient had failure-to-thrive as a child but had no history of pancreatitis. He had no mutations in LPL, APOA5, or APOC2. The Q115P substitution did not affect the ability of GPIHBP1 to reach the cell surface. However, unlike wild-type GPIHBP1, GPIHBP1-Q115P lacked the ability to bind LPL or chylomicrons (d < 1.006 g/mL lipoproteins from Gpihbp1(-/-) mice). Mouse GPIHBP1 with the corresponding mutation (Q114P) also could not bind LPL. CONCLUSIONS: A homozygous missense mutation in GPIHBP1 (Q115P) was identified in a patient with chylomicronemia. The mutation eliminated the ability of GPIHBP1 to bind LPL and chylomicrons, strongly suggesting that it caused the patient's chylomicronemia. PMID- 19304574 TI - SR-BI selective lipid uptake: subsequent metabolism of acute phase HDL. AB - OBJECTIVE: The purpose of this study was to investigate the interaction of SAA and SR-BI in remodeling of acute phase HDL (AP HDL). METHODS AND RESULTS: We used SAA and SR-BI adenoviral vector expression models to study the interaction between these entities. SR-BI processing of mouse AP HDL generated progressively smaller discreet HDL particles with distinct apolipoprotein compositions. SR-BI actions segregated apolipoproteins with the smallest particles containing only apoA-I. Larger remnants contained apoA-I, apoA-II, and SAA. Small apoA-I only particles failed to associate with preformed HDL, whereas larger remnants readily did. The presence of SAA on SR-BI-processed HDL particles propelled apoA-I to a small lipid-poor form and accelerated apoA-I catabolism. CONCLUSIONS: Data indicate that after core and surface HDL lipid perturbation by SR-BI, SAA propels apoA-I to a small lipid-poor form while accelerating HDL metabolism. PMID- 19304575 TI - Calcification by valve interstitial cells is regulated by the stiffness of the extracellular matrix. AB - OBJECTIVE: Extensive remodeling of the valve ECM in calcific aortic valve sclerosis alters its mechanical properties, but little is known about the impact of matrix mechanics on the cells within the valve interstitium. In this study, the influence of matrix stiffness in modulating calcification by valve interstitial cells (VICs), and their differentiation to pathological phenotypes was assessed. METHODS AND RESULTS: Primary porcine aortic VICs were cultured in standard media or calcifying media on constrained type I fibrillar collagen gels. Matrix stiffness was altered by changing only the thickness of the gels. Calcification did not occur in standard media, regardless of matrix stiffness. However, when VICs were grown in calcifying media on relatively compliant matrices with stiffness similar to that of normal tissue, they readily formed calcified aggregates of viable cells that expressed osteoblast-related transcripts and proteins. In contrast, VICs cultured in calcifying media on stiffer matrices (similar to stenotic tissue) differentiated to myofibroblasts and formed calcified aggregates that contained apoptotic cells. Actin depolymerization reduced aggregation on stiff, but not compliant, matrices. TGF beta1 potentiated aggregate formation on stiff matrices by enhancing alpha-smooth muscle actin expression and cellular contractility, but not on compliant matrices attributable to downregulation of TGF-beta receptor I. Cell contraction by VICs inhibited Akt activation and enhanced apoptosis-dependent calcification on stiff matrices. CONCLUSIONS: Differentiation of VICs to pathological phenotypes in response to biochemical cues is modulated by matrix stiffness. Although osteogenic or myofibrogenic differentiation of VICs can result in calcification, the processes are distinct. PMID- 19304576 TI - Rab8 regulates ABCA1 cell surface expression and facilitates cholesterol efflux in primary human macrophages. AB - OBJECTIVE: ATP-binding cassette transporter A1 (ABCA1) is thought to lipidate apolipoprotein A-I (apoA-I) at the plasma membrane, with endosomal cholesterol contributing as substrate. The mechanisms of ABCA1 surface delivery are not well understood. We have shown that Rab8 regulates endosomal cholesterol removal to apoA-I in human fibroblasts. Here, we investigated whether Rab8 plays a role in ABCA1 plasma membrane expression and cholesterol removal in primary human macrophages. METHODS AND RESULTS: We found that Rab8 was abundantly expressed in human atherosclerotic lesional macrophages and upregulated on lipid loading of macrophages in vitro. Adenoviral overexpression of Rab8 increased ABCA1 protein levels and reduced cholesterol deposition in macrophage foam cells incubated with apoA-I. Depletion of Rab8 decreased the fraction of ABCA1 at the plasma membrane and inhibited the efflux of lipoprotein-derived endosomal cholesterol to apoA-I. In Rab8-depleted cells, ABCA1-GFP localized in beta1 integrin and transferrin receptor containing recycling organelles. CONCLUSIONS: Rab8 reduces foam cell formation by facilitating ABCA1 surface expression and stimulating endosomal cholesterol efflux to apoA-I in primary human macrophages. PMID- 19304577 TI - Larval therapy for leg ulcers (VenUS II): randomised controlled trial. AB - OBJECTIVE: To compare the clinical effectiveness of larval therapy with a standard debridement technique (hydrogel) for sloughy or necrotic leg ulcers. DESIGN: Pragmatic, three armed randomised controlled trial. SETTING: Community nurse led services, hospital wards, and hospital outpatient leg ulcer clinics in urban and rural settings, United Kingdom. PARTICIPANTS: 267 patients with at least one venous or mixed venous and arterial ulcer with at least 25% coverage of slough or necrotic tissue, and an ankle brachial pressure index of 0.6 or more. INTERVENTIONS: Loose larvae, bagged larvae, and hydrogel. MAIN OUTCOME MEASURES: The primary outcome was time to healing of the largest eligible ulcer. Secondary outcomes were time to debridement, health related quality of life (SF-12), bacterial load, presence of meticillin resistant Staphylococcus aureus, adverse events, and ulcer related pain (visual analogue scale, from 0 mm for no pain to 150 mm for worst pain imaginable). RESULTS: Time to healing was not significantly different between the loose or bagged larvae group and the hydrogel group (hazard ratio for healing using larvae v hydrogel 1.13, 95% confidence interval 0.76 to 1.68; P=0.54). Larval therapy significantly reduced the time to debridement (2.31, 1.65 to 3.2; P<0.001). Health related quality of life and change in bacterial load over time were not significantly different between the groups. 6.7% of participants had MRSA at baseline. No difference was found between larval therapy and hydrogel in their ability to eradicate MRSA by the end of the debridement phase (75% (9/12) v 50% (3/6); P=0.34), although this comparison was underpowered. Mean ulcer related pain scores were higher in either larvae group compared with hydrogel (mean difference in pain score: loose larvae v hydrogel 46.74 (95% confidence interval 32.44 to 61.04), P<0.001; bagged larvae v hydrogel 38.58 (23.46 to 53.70), P<0.001). CONCLUSIONS: Larval therapy did not improve the rate of healing of sloughy or necrotic leg ulcers or reduce bacterial load compared with hydrogel but did significantly reduce the time to debridement and increase ulcer pain. TRIAL REGISTRATION: Current Controlled Trials ISRCTN55114812 and National Research Register N0484123692. PMID- 19304581 TI - The variation in transparency of amniotic membrane used in ocular surface regeneration. AB - BACKGROUND/AIMS: Scant consideration has been given to the variation in structure of the human amniotic membrane (AM) at source or to the significance such differences might have on its clinical transparency. Therefore, we applied our experience of quantifying corneal transparency to AM. METHODS: Following elective caesarean, AM from areas of the fetal sac distal and proximal (ie, adjacent) to the placenta was compared with freeze-dried AM. The transmission of light through the AM samples was quantified spectrophotometrically; also, tissue thickness was measured by light microscopy and refractive index by refractometry. RESULTS: Freeze-dried and freeze-thawed AM samples distal and proximal to the placenta differed significantly in thickness, percentage transmission of visible light and refractive index. The thinnest tissue (freeze-dried AM) had the highest transmission spectra. The thickest tissue (freeze-thawed AM proximal to the placenta) had the highest refractive index. Using the direct summation of fields method to predict transparency from an equivalent thickness of corneal tissue, AM was found to be up to 85% as transparent as human cornea. CONCLUSION: When preparing AM for ocular surface reconstruction within the visual field, consideration should be given to its original location from within the fetal sac and its method of preservation, as either can influence corneal transparency. PMID- 19304578 TI - Cost effectiveness analysis of larval therapy for leg ulcers. AB - OBJECTIVE: To assess the cost effectiveness of larval therapy compared with hydrogel in the management of leg ulcers. DESIGN: Cost effectiveness and cost utility analyses carried out alongside a pragmatic multicentre, randomised, open trial with equal randomisation. Population Intention to treat population comprising 267 patients with a venous or mixed venous and arterial ulcers with at least 25% coverage of slough or necrotic tissue. INTERVENTIONS: Patients were randomly allocated to debridement with bagged larvae, loose larvae, or hydrogel. MAIN OUTCOME MEASURE: The time horizon was 12 months and costs were estimated from the UK National Health Service perspective. Cost effectiveness outcomes are expressed in terms of incremental costs per ulcer-free day (cost effectiveness analysis) and incremental costs per quality adjusted life years (cost utility analysis). RESULTS: The larvae arms were pooled for the main analysis. Treatment with larval therapy cost, on average, pound96.70 (euro109.61; $140.57) more per participant per year (95% confidence interval - pound491.9 to pound685.8) than treatment with hydrogel. Participants treated with larval therapy healed, on average, 2.42 days before those in the hydrogel arm (95% confidence interval 0.95 to 31.91 days) and had a slightly better health related quality of life, as the annual difference in QALYs was 0.011 (95% confidence interval -0.067 to 0.071). However, none of these differences was statistically significant. The incremental cost effectiveness ratio for the base case analysis was estimated at pound8826 per QALY gained and pound40 per ulcer-free day. Considerable uncertainty surrounds the outcome estimates. CONCLUSIONS: Debridement of sloughy or necrotic leg ulcers with larval therapy is likely to produce similar health benefits and have similar costs to treatment with hydrogel. TRIAL REGISTRATION: Current Controlled Trials ISRCTN55114812 and National Research Register N0484123692. PMID- 19304582 TI - Comparison of clinical and photographic assessment of trachoma. AB - AIMS: The aim of the study was to determine the rates of trachoma in Aboriginal communities and to compare clinical assessment with photographic assessment for the presence of signs of trachoma. METHODS: Five Aboriginal communities in the Katherine region of the Northern Territory, Australia, were assessed for the presence of trachoma. Trachoma was diagnosed by clinical eye examination using a fine grading based on the World Health Organization (WHO) simplified grading system. Photographs were taken of the left eye of every person and graded using the fine grading system. The clinical assessment was compared with the photographic assessment for each person using the fine grading system. RESULTS: A total of 1316 people out of 1545 (85.2%) were screened for trachoma from five communities, with 1254 photographs being compared with clinical assessment scores. The overall prevalence of active trachoma was greater than 10% across the five communities, and greater than 20% in two communities. CONCLUSION: Active trachoma in young people and scarring in older people remain as problems in Aboriginal communities. Photographic assessment is a useful technique, but in comparison with clinical assessment it can result in overestimation of scoring for trachoma for inflammation. PMID- 19304583 TI - Subclinical keratoconus and inflammatory molecules from tears. AB - BACKGROUND/AIMS: Tissue degradation in corneal thinning disorders, such as keratoconus (KC), involves the expression of inflammatory mediators. The purpose of this study was to determine the levels of proinflammatory cytokines and matrix metalloproteinase 9 (MMP-9) in tears from both eyes of unilateral keratoconus (KC) patients. METHODS: Thirty patients diagnosed as having asymmetrical KC (30 KC eyes, and 30 subclinical KC eyes) and 20 normal control subjects (one eye) were studied in a prospective, cross-sectional study. Keratoconus screening programmes were performed on these participants. Ten microlitres of tears was collected from each eye. The concentrations of cytokines (interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-alpha)) and MMP-9 were measured by ELISA. RESULTS: Mean values for IL-6 levels were similar in KC and subclinical KC samples (5.5 (4.9 to 6.9) vs 5.7 (4.5 to 6.2) pg/ml, p = 0.131), but significantly higher in relation to the control group (2.2 (1.0 to 4.1) pg/ml, p<0.0001). Significant differences were found in TNF-alpha levels between KC and subclinical KC eyes (5.4 (4.1 to 6.8) vs 4.8 (4.2 to 6.0) pg/ml, p = 0.032) and control group (1.8 (1.5 to 2.3) pg/ml, p<0.0001). Increased values of MMP-9 were found in KC (59.4 (50.6 to 66.1) ng/ml) vs subclinical KC eye (7.0 (4.8 to 8.6) ng/ml) (p<0.0001). MMP-9 levels in the control group (6.1 (3.9 to 8.3) ng/ml) and subclinical KC were similar (p = 0.203). CONCLUSIONS: IL-6 and TNF-alpha are overexpressed in the tears of subclinical and KC eyes. Increased MMP-9 levels were found only in the KC eye. These results indicate that the pathogenesis of KC may involve chronic inflammatory events. PMID- 19304584 TI - Polypoidal choroidal vasculopathy and late geographic hyperfluorescence on indocyanine green angiography. AB - BACKGROUND/AIMS: To report the clinical significance of late geographic hyperfluorescence (LGH) on indocyanine green angiography (ICGA) in cases of polypoidal choroidal vasculopathy (PCV). METHODS: The medical records of 43 eyes with PCV, all of which had undergone at least 12 months of follow-up, 40 eyes with exudative age-related macular degeneration (AMD) and 20 eyes of age-matched normal subjects were retrospectively analysed. The incidence of LGH, defined as a well-demarcated geographic hyperfluorescent lesion on late phase ICGA, was compared in each respective group. The natural course of the LGH and its changes after photodynamic therapy (PDT) were analysed. RESULTS: LGH was noted in all of the eyes with PCV, whereas LGH was noted in three eyes (7.5%) of the eyes with exudative AMD and was not noted in any of the normal subjects (p<0.01). Of the 27 eyes (62.8%) with PCV, LGH was matched to the total area of the branching vascular network and polyps. The extent of LGH was enlarged over time in approximately one-half of the cases. As compared with the eyes demonstrating persistent LGH after PDT, the eyes with fading or disappearing LGH evidenced a lower recurrence of active PCV (p<0.05). CONCLUSION: LGH is a highly sensitive and specific ICGA finding for the diagnosis of PCV. Increased surveillance should be implemented in eyes in which LGH persists after PDT. PMID- 19304585 TI - Differences in retinal neovascular tissue and vitreous humour in patients with type 1 and type 2 diabetes. AB - AIMS: The aim of the study was to evaluate the histopathology of neovascular tufts and vitreous samples collected from patients with diabetes. METHODS: Vitreous samples and neovascular tufts were collected from patients with type 1 (n = 13) and (n = 17) type 2 diabetes with proliferative retinopathy, and from controls with a macular hole (n = 5). Neovessels were analysed using immunohistochemistry and vitreous samples with an enzyme-linked immunosorbent assay (ELISA). The main outcome measure was to examine differences in the levels of growth factors in patients with type 1 and type 2 diabetes with proliferative retinopathy. RESULTS: Vascular endothelial growth factor (VEGF)-A was most strongly present in the samples from patients with type 1 diabetes. In type 2 diabetes, VEGF-D was more abundantly present than in type 1 diabetes. Angiopoietin (ANG)-2 was also abundantly present. Macrophages and nuclear factor kappa B (NFkappaB) were found, indicating the presence of an inflammatory process in the neovascular tissues. CONCLUSIONS: VEGF-A and ANG-2 are equally important in the neovascular process in both type 1 and type 2 diabetes. VEGF-D is abundantly present in type 2 diabetes. In order to achieve better control of diabetic retinopathy, it might be beneficial to develop treatments that prevent the actions of ANG-2 and VEGF-D. PMID- 19304586 TI - Agreement between spectral-domain and time-domain OCT for measuring RNFL thickness. AB - BACKGROUND/AIMS: To evaluate spectral-domain (SD) optical coherence tomography (OCT) reproducibility and assess the agreement between SD-OCT and Time-Domain (TD) OCT retinal nerve fibre layer (RNFL) measurements. METHODS: Three Cirrus-SD OCT scans and one Stratus-TD-OCT scan were obtained from Diagnostic Innovations in Glaucoma Study (DIGS) healthy participants and glaucoma patients on the same day. Repeatability was evaluated using Sw (within-subject standard deviation), CV (coefficient of variation) and ICC (intraclass correlation coefficient). Agreement was assessed using correlation and Bland-Altman plots. RESULTS: 16 healthy participants (32 eyes) and 39 patients (78 eyes) were included. SD-OCT reproducibility was excellent in both groups. The CV and ICC for Average RNFL thickness were 1.5% and 0.96, respectively, in healthy eyes and 1.6% and 0.98, respectively, in patient eyes. Correlations between RNFL parameters were strong, particularly for average RNFL thickness (R(2) = 0.92 in patient eyes). Bland Altman plots showed good agreement between instruments, with better agreement for average RNFL thickness than for sectoral RNFL parameters (for example, at 90 microm average RNFL thickness, 95% limits of agreement were -13.1 to 0.9 for healthy eyes and -16.2 to -0.3 microm for patient eyes). CONCLUSIONS: SD-OCT measurements were highly repeatable in healthy and patient eyes. Although the agreement between instruments was good, TD-OCT provided thicker RNFL measurements than SD-OCT. Measurements with these instruments should not be considered interchangeable. PMID- 19304587 TI - Expression of reverse cholesterol transport proteins ATP-binding cassette A1 (ABCA1) and scavenger receptor BI (SR-BI) in the retina and retinal pigment epithelium. AB - AIMS: Excessive lipid accumulation in Bruch's membrane (BrM) is a hallmark of ageing, the major risk factor for age-related macular degeneration (AMD). Retinal pigment epithelial (RPE) cells may utilise reverse cholesterol transport (RCT) activity to move lipid into BrM, mediated through ATP-binding cassette A1 (ABCA1) and scavenger receptor BI (SR-BI). METHODS: ABCA1 expression was assessed by reverse transcription polymerase chain reaction (RT-PCR) and western blotting of human RPE cell extracts. Lipid transport assays were performed using radiolabelled photoreceptor outer segments (POS). ABCA1 and SR-BI expression was examined in normal mouse eyes by immunofluorescence staining. BrMs of ABCA1 and SR-BI heterozygous mice were examined microscopically. RESULTS: Human RPE cells expressed ABCA1 mRNA and protein. The ABCA1 and SR-BI inhibitor glyburide (also known as glibenclamide) abolished basal transport of POS-derived lipids in RPE cells in the presence of high-density lipoprotein. Mouse retina and RPE expressed ABCA1 and SR-BI. SR-BI was highly expressed in RPE. BrMs were significantly thickened in SR-BI heterozygous mice, but not in ABCA1 heterozygous mice. CONCLUSION: RPE cells express ABCA1 and SR-BI. This implies a significant role for SR-BI and ABCA1 in lipid transport and RCT in the retina and RPE. PMID- 19304588 TI - Three-dimensional reconstruction of sentinel lymph nodes with metastatic breast cancer indicates three distinct patterns of tumour growth. AB - AIM: A better understanding of the biology of nodal metastatic disease is of indisputable value. Three-dimensional (3D) serial section alignment and reconstruction techniques can be used for visualisation of nodal metastasis and could provide better understanding of disease growth patterns. METHODS: 19 tumour involved sentinel nodes (SLNs) from breast cancer patients were serially sectioned, immunohistochemically stained, and digitally scanned. Digital image alignment and voxel-based rendering was used to construct informative 3D visual representations of metastatic tumour distribution within involved nodes. RESULTS: The 3D reconstruction technique was successful and informative. The reconstructions of all 19 SLNs enabled the metastatic tumour cells to be viewed infiltrating normal SLN tissue from all angles. Metastases were present at the afferent lymphatic pole in 17/19 cases, confined to the afferent pole only in 7 cases, located at the efferent pole in 12/19 cases, and efferent pole only in just 2 cases. Finally, this study made the novel observation that metastatic growth occurs in three distinct patterns: sinusoidal, nodular and diffuse. CONCLUSIONS: This methodology provides improved understanding of metastatic disease development and potentially could be used to develop strategies to improve techniques for its routine detection. Further studies are required in order to evaluate the prognostic and biological significance of the growth patterns identified. PMID- 19304589 TI - Patterns of intraspecific DNA variation in the Daphnia nuclear genome. AB - Understanding nucleotide variation in natural populations has been a subject of great interest for decades. However, many taxonomic groups, especially those with atypical life history attributes remain unstudied, and Drosophila is the only arthropod genus for which DNA polymorphism data are presently abundant. As a result of the recent release of the complete genome sequence and a wide variety of new genomic resources, the Daphnia system is quickly becoming a promising new avenue for expanding our knowledge of nucleotide variation in natural populations. Here, we examine nucleotide variation in six protein-coding loci for Daphnia pulex and its congeners with particular emphasis on D. pulicaria, the closest extant relative of D. pulex. Levels of synonymous intraspecific variation, pi(s), averaged 0.0136 for species in the Daphnia genus, and are slightly lower than most prior estimates in invertebrates. Tests of neutrality indicated that segregating variation conforms to neutral model expectations for the loci that we examined in most species, while K(a)/K(s) ratios revealed strong purifying selection. Using a full maximum-likelihood coalescent-based method, the ratio of the recombination rate to the mutation rate (c/u), averaged 0.5255 for species of the Daphnia genus. Lastly, a divergence population-genetics approach was used to investigate gene flow and divergence between D. pulex and D. pulicaria. PMID- 19304592 TI - Sensory noise explains auditory frequency discrimination learning induced by training with identical stimuli. AB - Thresholds in various visual and auditory perception tasks have been found to improve markedly with practice at intermediate levels of task difficulty. Recently, however, there have been reports that training with identical stimuli, which, by definition, were impossible to discriminate correctly beyond chance, could induce as much discrimination learning as could training with different stimuli. These surprising findings have been interpreted as evidence that discrimination learning can occur in the absence of perceived differences between stimuli and need not involve the fine-tuning of a discrimination mechanism. Here, we show that these counterintuitive findings of discrimination learning without discrimination can be understood simply by considering the effect of internal noise on sensory representations. Because of such noise, physically identical stimuli are unlikely to be perceived as being strictly identical. We show that, given empirically derived levels of sensory noise, perceived differences evoked by identical stimuli are actually not much smaller than those induced by the physical differences typically used in discrimination-learning experiments. We suggest that findings of discrimination learning with identical stimuli can be explained without implicating any fundamentally new learning mechanism. PMID- 19304593 TI - Symmetrical interaction of sex and expression in face classification tasks. AB - Classification of faces as to their sex or their expression--with sex and expression varying orthogonally--was studied in three experiments. In Experiment 1, expression classification was influenced by sex, with angry male faces being classified faster than angry female faces. Complementarily, sex classification was faster for happy than for angry female faces. In Experiment 2, mutual interaction of sex and expression was also found when the participants were asked to classify top and bottom face segments. In Experiment 3, a face inversion effect was found for both sex and expression classification of whole faces. However, a symmetrical interaction between sex and expression was again found. The results are discussed in terms of configural versus feature processing in the perception of face sex and expression and of their relevance to face perception models that postulate independent processing of different facial features. PMID- 19304594 TI - The effects of implicit attentional learning and habituation on inhibition of return. AB - In two experiments, the effects of implicit attentional learning and habituation on inhibition of return (IOR) were assessed. Experiment 1 consisted of an informative variant of the spatial cuing paradigm (Posner, 1980), with the target appearing most often at the location next to the cue. Short-term as well as long term learning of the regularities could be observed. This learning took place even though the participants were not able to verbalize the learned information, suggesting that the learning was implicit. By spatially separating the effects of implicit learning and IOR, we demonstrated that the magnitude of IOR slightly decreased because of long-term implicit orienting. In Experiment 2, a noninformative variant of the spatial cuing paradigm, IOR was still found to decrease over the course of the experiment. This suggests that habituation due to the mere repeated initiation of IOR also affects the magnitude of the IOR. This theory is supported by the finding that IOR recovered after short breaks in Experiment 2. However, habituation presumably only played a secondary role in Experiment 1. In sum, IOR was abolished by neither implicit attentional learning nor habituation. Here, the finding that IOR is not easily replaced by implicitly learned orienting clearly indicates that IOR is a powerful, low-level mechanism. We discuss implications of these results for the debate on the function of IOR. PMID- 19304595 TI - Spatial updating in superimposed real and virtual environments. AB - Wang (2004) showed that people do not always simultaneously update their relationships to real and imagined environments in a dual-environment situation. Employing the same paradigm, we examined whether spatial updating operates on virtual reality as it does on a real or fictitious environment. Participants learned target locations in a real room and a virtual kitchen. Then they turned to face targets either in the room or in the kitchen, while blindfolded, and pointed to the targets before and after turning. Participants kept track of their orientation in both environments equally efficiently, regardless of explicit instructions. In contrast, when the real environment was described verbally but not directly perceived, participants automatically updated the virtual kitchen but not the room. These results suggest that people automatically update a virtual environment as they do a real one when the two environments are superimposed. The automaticity of spatial updating is discussed. PMID- 19304596 TI - Relations between uniform connectedness, luminance, and shape similarity as perceptual organizational cues in infancy. AB - Although several studies have examined infants' sensitivity to perceptual organizational cues, few have examined the functional relations among these cues. We examined how uniform connectedness (UC) functions in relation to shape and luminance similarity. UC has been characterized as the entry-level mechanism of perceptual organization and would therefore be predicted to be more salient than the other two cues. We found that UC was more salient than shape similarity organization was, to the point that 6- to 7-month-old infants failed to even organize on the basis of shape in the presence of UC. Luminance similarity, however, was more salient than UC, even though UC was detected by infants in the presence of luminance cues. We conclude that UC is not necessarily the most salient mechanism of perceptual organization in infancy. Moreover, the luminance UC-shape salience hierarchy exhibited by 6- to 7-month-olds in the present study is consistent with the order of development of sensitivity to these organizational cues. PMID- 19304597 TI - Individual differences in learning to perceive length by dynamic touch: evidence for variation in perceptual learning capacities. AB - Recent studies of perceptual learning have explored and commented on variation in learning trajectories. Although several factors have been suggested to account for this variation, thus far the idea that humans vary in their perceptual learning capacities has received scant attention. In the present experiment, we aimed at providing a detailed picture of the variation in this capacity by investigating the perceptual learning trajectories of a considerable number of participants. The learning process was studied using the paradigm of length perception by dynamic touch. The results showed that there are substantial individual differences in the way perceivers respond to feedback. Indeed, after feedback, the participants' perceptual performances diverged. We conclude that humans vary in their perceptual learning capacities. The implications of this finding for recent discussions on variation in perception are explored. PMID- 19304598 TI - The role of peripheral vision in implicit contextual cuing. AB - Implicit contextual cuing refers to the ability to learn the association between contextual information of our environment and a specific target, which can be used to guide attention during visual search. It was recently suggested that the storage of a snapshot image of the local context of a target underlies implicit contextual cuing. To make such a snapshot, it is necessary to use peripheral vision. In order to test whether peripheral vision can underlie implicit contextual cuing, we used a covert visual search task, in which participants were required to indicate the orientation of a target stimulus while foveating a fixation cross. The response times were shorter when the configuration of the stimuli was repeated than when the configuration was new. Importantly, this effect was still found after 10 days, indicating that peripherally perceived spatial context information can be stored in memory for long periods of time. These results indicate that peripheral vision can be used to make a snapshot of the local context of a target. PMID- 19304599 TI - Comparison of perceptual and motor latencies via anticipatory and reactive response times. AB - To compare the timing of perceptual and motor decisions, distinct tasks have been designed, all of which have yielded systematic differences between these two moments. These observations have been taken as evidence of a sensorimotor dissociation. Inasmuch as the distinction between perceptual and motor decision moments is conceptually warranted, this conclusion remains debatable, since the observed differences may reflect the dissimilarity between the stimulations/tasks used to assess them. Here, we minimize such dissimilarities by comparing response time (RT) and anticipatory RT (ART), an alternative technique with which to infer the relative perceptual decision moments. Observers pressed a key either in synchrony with the third of a sequence of three stimuli appearing at a constant pace (ART) or in response to the onset of this third stimulus presented at a random interval after the second (RT). Hence, the two stimulation sequences were virtually identical. Both the mean and the variance of RT were affected by stimulus intensity about 1.5 times more than were the mean and the variance of ART. Within the framework of two simple integration-to-bound models, these findings are compatible with the hypothesis that perceptual and motor decisions operate on the same internal signal but are based on distinct criteria, with the perceptual criterion lower than the motor one. PMID- 19304600 TI - Unconscious processing of color and form in metacontrast masking. AB - Three experiments employed a metacontrast masking procedure to examine the extent and nature of priming effects from visual stimuli not consciously perceived. The results showed effects of unconscious stimuli on subsequent target responses that (1) were more consistent, reliable, and not subject to strategic control, as compared with consciously perceived stimuli (Experiment 1); (2) produced both facilitation and interference of subsequent processing (Experiment 2); and (3) did not influence indirect response-related levels of processing (Experiment 3). These results demonstrate that color and form attributes of unconscious stimuli are sufficiently registered within the visual system to influence behavior, and that some of these unconscious effects occur at early levels of stimulus encoding, prior to higher level perceptual and response-related processes. PMID- 19304601 TI - Perceived range, perceived velocity, and perceived duration of the body rotating in the frontal plane. AB - We investigated perceived range, perceived velocity, and perceived duration of the body rotating in the frontal plane (in roll). Specifically, to examine how shear to the otoliths in the inner ears and tactile pressure to the trunk affect judgments of range and velocity, in two experiments, we manipulated rotating range (30 degrees-160 degrees), rotating velocity (1.8 degrees/sec to 9.6 degrees/sec), mean tilt of the body (-60 degrees, 0 degrees, and 60 degrees), and exposure to the visual vertical. Thirty-three normal or blindfolded participants made verbal judgments of range, velocity, and duration for each combination of these factors. The exponents of the power functions fitted to these judgments were, as a first approximation, .94, .61, and .84 for range, velocity, and duration, respectively, and perceived velocity was proportional to the ratio of perceived range to perceived duration (r = .91). These results suggest that the vestibular and somatosensory inputs are effective on judgments of range, but less so on judgments of velocity, and that perceived velocity may be determined as a ratio of perceived range to perceived duration. In addition, we found that (1) when the range the body has traveled is constant, the perceived range increases as the objective velocity decreases (proprioceptive tau effect); (2) self-motion through the tilted roll sometimes enlarges perceived range and perceived duration but reduces perceived velocity; and (3) the exposure to the visual vertical reduces variability of judgments for range and velocity and also reduces perceived range and perceived velocity of self-motion within a small range through the vertical roll. PMID- 19304602 TI - Aging and the perception of slant from optical texture, motion parallax, and binocular disparity. AB - The ability of younger and older observers to perceive surface slant was investigated in four experiments. The surfaces possessed slants of 20 degrees, 35 degrees, 50 degrees, and 65 degrees, relative to the frontoparallel plane. The observers judged the slants using either a palm board (Experiments 1, 3, and 4) or magnitude estimation (Experiment 2). In Experiments 1-3, physically slanted surfaces were used (the surfaces possessed marble, granite, pebble, and circle textures), whereas computer-generated 3-D surfaces (defined by motion parallax and binocular disparity) were utilized in Experiment 4. The results showed that the younger and older observers' performance was essentially identical with regard to accuracy. The younger and older age groups, however, differed in terms of precision in Experiments 1 and 2: The judgments of the older observers were more variable across repeated trials. When taken as a whole, the results demonstrate that older observers (at least through the age of 83 years) can effectively extract information about slant in depth from optical patterns containing texture, motion parallax, or binocular disparity. PMID- 19304603 TI - The horizon line, linear perspective, interposition, and background brightness as determinants of the magnitude of the pictorial moon illusion. AB - A total of 110 undergraduate students participated in a series of three experiments that explored the magnitude of the moon illusion in pictures. Experiment 1 examined the role of the number and salience of depth cues and background brightness. Experiment 2 examined the role of the horizon line, linear perspective, interposition, and background brightness. In Experiment 3, comparative distance judgments of the moon as a function of linear perspective, interposition, and the size of the standard moon were obtained. The magnitude of the moon illusion increased as a function of the number and salience of depth cues and changes in background brightness. Experiment 2 failed to support the role of the horizon line in affecting the illusion. Experiment 3 provided additional support for the illusory distance component of the moon illusion. PMID- 19304604 TI - The role of parity, physical size, and magnitude in numerical cognition: the SNARC effect revisited. AB - People indicate the physical size or the parity status of small numbers faster by a left-hand key and those of larger numbers by a right-hand key. Because magnitude information is not required for successful performance in these tasks, the presence of a number-space association (the SNARC effect) has been taken to indicate the automatic activation of numerical magnitude in all tasks with numerals. In order to test this account, in a series of five experiments, we derived two consensual markers of automatic activation of irrelevant numerical magnitude, the size congruity effect (for judgments of physical size), and the Garner effect (for judgments of parity). Both markers were found independent of the SNARC effect. Consequently, we question the traditional explanation of the SNARC effect and offer an alternative account in terms of a highly overlearned stimulus-response loop. PMID- 19304605 TI - Spatial acuity and summation on the hand: the role of thermal cues in material discrimination. AB - The spatial characteristics of thermal perception were studied in two experiments that examined how thermal stimuli are processed within the hands. A thermal display that simulates cues associated with making contact with different materials was used in these studies. In the first experiment, participants indicated which of two simulated materials that were presented to the index fingertip was cooler. The results indicated that participants were unable to resolve the two areas of thermal stimulation. In the second experiment, the effects of concurrent thermal stimulation on the ability to discriminate between simulated materials were evaluated. Thermal cues were presented to the middle fingers of both hands and to two adjacent fingers on one hand. Thermal spatial summation was evident across the fingers, which enhanced the ability to discriminate between materials when the cooler stimulus was presented to three fingers. When the same stimulus was presented to the two hands, the stimulation of adjacent fingers altered the perceived thermal response. PMID- 19304606 TI - Speech intelligibility reduces over distance from an attended location: evidence for an auditory spatial gradient of attention. AB - Speech reception thresholds (SRTs) were measured at a central focus of attention and at 20 degrees, 40 degrees, and 60 degrees locations distant in azimuth. Measurements were taken with one target collocated with two maskers, or with maskers flanking the target by +/-20 degrees. For 80% of trials, the target was played from the attended location ("expected"), and 20% came from another ("unexpected") location. For collocated stimuli, SRTs worsened with increasing distance from the expected location by 2.1 dB over the 60 degrees azimuth and by 5.1 dB for spatially separated target and maskers. In spatially separated conditions, a 2.9-dB change was still found when gaze was away from the attended location. Spatial attention appears to increase speech intelligibility against interferers, with gain decreasing with distance from the focus of attention. Spatial release from masking (RFM) was only found for the attended location. Further experiments suggest that target location uncertainty prevented RFM at unattended locations. PMID- 19304607 TI - Conditions required for binocular rivalry suppression. AB - When the two eyes are presented with incompatible stimuli, the two monocular stimuli are seen alternately in a never-ending cycle. It is now widely accepted that the neural processes underlying this phenomenon, binocular rivalry, are distributed across a number of cortical stages. It is not clear, however, where binocular rivalry is initiated. We performed two experiments whose aim was to clarify this issue. In the first experiment, rivalry was induced, and brief test stimuli were delivered to an eye while its inducing stimulus was either dominant or suppressed. Sensitivity to a test stimulus with features similar to those of the suppressed inducing stimulus was reduced only when the test was presented to the eye whose inducing stimulus was suppressed. This indicates that suppression of a monocular channel is a prerequisite for binocular rivalry suppression. The second experiment showed that to induce rivalry, local interocular stimulus incompatibilities were necessary and that conflicting global percepts were not sufficient. These results suggest that low-level visual processes are required for the initiation of binocular rivalry. PMID- 19304608 TI - The effect of the observer vantage point on perceived distortions in linear perspective images. AB - Some features of linear perspective images may look distorted. Such distortions appear in two drawings by Jan Vredeman de Vries involving perceived elliptical, instead of circular, pillars and tilted, instead of upright, columns. Distortions may be due to factors intrinsic to the images, such as violations of the so called Perkins's laws, or factors extrinsic to them, such as observing the images from positions different from their center of projection. When the correct projection centers for the two drawings were reconstructed, it was found that they were very close to the images and, therefore, practically unattainable in normal observation. In two experiments, enlarged versions of images were used as stimuli, making the positions of the projection centers attainable for observers. When observed from the correct positions, the perceived distortions disappeared or were greatly diminished. Distortions perceived from other positions were smaller than would be predicted by geometrical analyses, possibly due to flatness cues in the images. The results are relevant for the practical purposes of creating faithful impressions of 3-D spaces using 2-D images. PMID- 19304609 TI - The development of depth perception from motion parallax in infancy. AB - Little is known about infants' perception of depth from motion parallax, even though it is known that infants are sensitive both to motion and to depth-from motion cues at an early age. The present experiment assesses whether infants are sensitive to the unambiguous depth specified by motion parallax and, if so, when this sensitivity first develops. Eleven infants were followed longitudinally from 8 to 29 weeks. Infants monocularly viewed a translating Rogers and Graham (1979) random-dot stimulus, which appears as a corrugated surface to adult observers. Using the infant-control habituation paradigm, looking time was recorded for each 10-sec trial until habituation, followed by two test trials: one using a depth reversed and one using a flat stimulus. Dishabituation results indicate that infants may be sensitive to unambiguous depth from motion parallax by 16 weeks of age. Implications for the developmental sequence of depth from motion, stereopsis, and eye movements are discussed. PMID- 19304610 TI - Shape memory for intrinsic versus accidental holes. AB - Two experiments were performed to investigate the circumstances under which the shape of a visual hole is perceived and remembered. A distinction is made between an intrinsic hole, a surrounded region that is bordered on all sides by a single surface at a continuous depth, and an accidental hole, a surrounded region with borders belonging to multiple surfaces at discontinuous depths. In the first experiment, we demonstrated a marked memory advantage for intrinsic holes over both accidental holes and accidental parts when the accidental hole and part were nested within the intrinsic hole. Indeed, the accidental holes and parts appeared not to be remembered at all. In Experiment 2, we used nonnested display sets that ruled out the possibility that the results from the first experiments were due to any image-based differences and included solid objects, as well as intrinsic and accidental holes. The results showed that the shapes of intrinsic holes were still remembered better than those of accidental holes, but not as well as those of solid objects. The results are discussed in a framework of ecological validity, arguing that observers are likely to encode shapes that are invariant features of the environmental scene and not those that arise only from the coincidence of particular viewing conditions. PMID- 19304611 TI - The effect of pictorial depth information on projected size judgments. AB - When full depth cues are available, size judgments are dominated by physical size. However, with reduced depth cues, size judgments are influenced less by physical size and more by projected size. By manipulating monocularly presented pictorial depth cues only, in this study we reduced depth cues further than had previous size judgment studies. Participants were presented monocularly with two shapes against a background of zero (control), one, two, or three pictorial depth cues. Each cue was added progressively in the following order: height in the visual field, linear perspective, and texture gradient. Participants made a same/different judgment regarding the projected size of the two shapes (i.e., ignoring any depth cues). As was expected, accuracy increased and response times decreased as the ratio between the projected size of the two shapes increased (range of projected size ratios, 1:1-1:5). In addition, with the exception of the larger size ratios (1:4 and 1:5), detection of projected size difference grew poorer as depth cues were added. One- and two-cue conditions had the most weighting in this performance decrement, with little weighting from the three-cue condition. We conclude that even minimal depth information is difficult to inhibit, which indicates that depth perception requires little focused attention. PMID- 19304613 TI - Foreshortened tiles in paths converging on an observer viewing a picture: elevation and visual angle ratio determine perceived size. AB - Observers were shown wide-angle pictures of tiles on a ground plane and were asked about the aspect ratios of the tiles. The observers viewed the pictures from a fixed center of projection. Some of the tiles were in a path coming straight toward the observer. In one picture, the path came from the center of the picture, and in two others the path came from the left side of the picture (one from 30 degrees and one from 45 degrees to the left of the center, from the observer's point of view). The apparent aspect ratios were a function of the elevations of the tiles and the ratios of visual angles of the sides of the tiles. Judgments were identical for all three paths. The local slant of the picture surface was not a significant factor. PMID- 19304614 TI - Understanding the underlying dimensions in perfumers' odor perception space as a basis for developing meaningful odor maps. AB - Various low-dimensional perceptual maps of fragrances have been proposed in the literature, as well as sensory maps for the odor descriptors most frequently applied in perfumery. To reach a consensus, however, seems difficult, if at all possible. In the present study, we applied principal components analysis to two databases. The first contains numeric odor profiles of 309 compounds based on 30 descriptors. The loading plot corresponding to the relevant components was strikingly similar to the odor effects diagram proposed by P. Jellinek (1951), primarily on the basis of his long experience as a perfumer. We obtained similar results in our analysis of the second database, which comprises 66 descriptors and contains the semantic descriptions of 119 perfume materials. On the basis of the results of both analyses, a commercial map of fragrances is discussed. Our findings suggest that it is possible to develop standard sensory maps of perfumery odor descriptors, if a consensus is first reached regarding which odorants best represent particular odor qualities. PMID- 19304615 TI - Logical recoding of S-R rules can reverse the effects of spatial S-R correspondence. AB - Two experiments investigated competing explanations for the reversal of spatial stimulus-response (S-R) correspondence effects (i.e., Simon effects) with an incompatible S-R mapping on the relevant, nonspatial dimension. Competing explanations were based on generalized S-R rules (logical-recoding account) or referred to display-control arrangement correspondence or to S-S congruity. In Experiment 1, compatible responses to finger-name stimuli presented at left/right locations produced normal Simon effects, whereas incompatible responses to finger name stimuli produced an inverted Simon effect. This finding supports the logical recoding account. In Experiment 2, spatial S-R correspondence and color S-R correspondence were varied independently, and main effects of these variables were observed. The lack of an interaction between these variables, however, disconfirms a prediction of the display-control arrangement correspondence account. Together, the results provide converging evidence for the logical recoding account. This account claims that participants derive generalized response selection rules (e.g., the identity or reversal rule) from specific S-R rules and inadvertently apply the generalized rules to the irrelevant (spatial) S R dimension when selecting their response. PMID- 19304616 TI - A feature-weighting account of priming in conjunction search. AB - Previous research on the priming effect in conjunction search has shown that repeating the target and distractor features across displays speeds mean response times but does not improve search efficiency: Repetitions do not reduce the set size effect-that is, the effect of the number of distractor items-but only modulate the intercept of the search function. In the present study, we investigated whether priming modulates search efficiency when a conjunctively defined target randomly changes between red and green. The results from an eyetracking experiment show that repeating the target across trials reduced the set size effect and, thus, did enhance search efficiency. Moreover, the probability of selecting the target as the first item in the display was higher when the target-distractor displays were repeated across trials than when they changed. Finally, red distractors were selected more frequently than green distractors when the previous target had been red (and vice versa). Taken together, these results indicate that priming in conjunction search modulates processes concerned with guiding attention to the target, by assigning more attentional weight to features sharing the previous target's color. PMID- 19304617 TI - Spatial attention does improve temporal discrimination. AB - It has recently been stated that exogenous attention impairs temporal-resolution tasks (Hein, Rolke, & Ulrich, 2006; Rolke, Dinkelbach, Hein, & Ulrich, 2008; Yeshurun, 2004; Yeshurun & Levy, 2003). In comparisons of performance on spatially cued trials versus neutral cued trials, the results have suggested that spatial attention decreases temporal resolution. However, when performance on cued and uncued trials has been compared in order to equate for cue salience, typically speed-accuracy trade-offs (SATs) have been observed, making the interpretation of the results difficult. In the present experiments, we aimed at studying the effect of spatial attention in temporal resolution while using a procedure to control for SATs. We controlled reaction times (RTs) by constraining the time to respond, so that response decisions would be made within comparable time windows. The results revealed that when RT was controlled, performance was impaired for cued trials as compared with neutral trials, replicating previous findings. However, when cued and uncued trials were compared, performance was actually improved for cued trials as compared with uncued trials. These results suggest that SAT effects may have played an important role in the previous studies, because when they were controlled and measured, the results reversed, revealing that exogenous attention does improve performance on temporal resolution tasks. PMID- 19304618 TI - A multistream model of visual word recognition. AB - Four experiments are reported that test a multistream model of visual word recognition, which associates letter-level and word-level processing channels with three known visual processing streams isolated in macaque monkeys: the magno dominated (MD) stream, the interblob-dominated (ID) stream, and the blob dominated (BD) stream (Van Essen & Anderson, 1995). We show that mixing the color of adjacent letters of words does not result in facilitation of response times or error rates when the spatial-frequency pattern of a whole word is familiar. However, facilitation does occur when the spatial-frequency pattern of a whole word is not familiar. This pattern of results is not due to different luminance levels across the different-colored stimuli and the background because isoluminant displays were used. Also, the mixed-case, mixed-hue facilitation occurred when different display distances were used (Experiments 2 and 3), so this suggests that image normalization can adjust independently of object size differences. Finally, we show that this effect persists in both spaced and unspaced conditions (Experiment 4)--suggesting that inappropriate letter grouping by hue cannot account for these results. These data support a model of visual word recognition in which lower spatial frequencies are processed first in the more rapid MD stream. The slower ID and BD streams may process some lower spatial frequency information in addition to processing higher spatial frequency information, but these channels tend to lose the processing race to recognition unless the letter string is unfamiliar to the MD stream--as with mixed-case presentation. PMID- 19304619 TI - Anchoring effects in the judgment of confidence: semantic or numeric priming? AB - Over the last decade, researchers have debated whether anchoring effects are the result of semantic or numeric priming. The present study tested both hypotheses. In four experiments involving a sensory detection task, participants first made a relative confidence judgment by deciding whether they were more or less confident than an anchor value in the correctness of their decision. Subsequently, they expressed an absolute level of confidence. In two of these experiments, the relative confidence anchor values represented the midpoints between the absolute confidence scale values, which were either explicitly numeric or semantic, nonnumeric representations of magnitude. In two other experiments, the anchor values were drawn from a scale modally different from that used to express the absolute confidence (i.e., nonnumeric and numeric, respectively, or vice versa). Regardless of the nature of the anchors, the mean confidence ratings revealed anchoring effects only when the relative and absolute confidence values were drawn from identical scales. Together, the results of these four experiments limit the conditions under which both numeric and semantic priming would be expected to lead to anchoring effects. PMID- 19304620 TI - Additivity of abrupt onset effects supports nonspatial distraction, not the capture of spatial attention. AB - In a recent article, Schreij, Owens, and Theeuwes (2008) reported that abruptly onsetting distractors produce costs in performance even when spatial-cuing effects confirm the presence of a top-down set for color. The authors argued that these results show that abruptly onsetting new objects capture attention independent of a top-down set and, thus, provide conclusive evidence against the theory that attentional capture is contingent on top-down attentional control settings (Folk, Remington, & Johnston, 1992). In the following article, we argue that, contrary to the conclusion drawn by Schreij et al., their own data (1) disconfirm the claim that their abrupt onsets captured spatial attention and (2) are consistent with nonspatial interference accounts of singleton-distractor effects. In support of the nonspatial account, we show that in a paradigm similar to Schreij et al.'s, distractors that do not capture attention can nonetheless influence responses to a target. We conclude that the results of Schreij et al. do not represent a challenge to contingent capture theory. PMID- 19304621 TI - Look away! Eyes and arrows engage oculomotor responses automatically. AB - The present study investigates how people's voluntary saccades are influenced by where another person is looking, even when this is counterpredictive of the intended saccade direction. The color of a fixation point instructed participants to make saccades either to the left or right. These saccade directions were either congruent or incongruent with the eye gaze of a centrally presented schematic face. Participants were asked to ignore the eyes, which were congruent only 20% of the time. At short gaze-fixation-cue stimulus onset asynchronies (SOAs; 0 and 100 msec), participants made more directional errors on incongruent than on congruent trials. At a longer SOA (900 msec), the pattern tended to reverse. We demonstrate that a perceived eye gaze results in an automatic saccade following the gaze and that the gaze cue cannot be ignored, even when attending to it is detrimental to the task. Similar results were found for centrally presented arrow cues, suggesting that this interference is not unique to gazes. PMID- 19304622 TI - Semisupervised category learning: the impact of feedback in learning the information-integration task. AB - In a standard supervised classification paradigm, stimuli are presented sequentially, participants make a classification, and feedback follows immediately. In this article, we use a semisupervised classification paradigm, in which feedback is given after a prespecified percentage of trials only. In Experiment 1, feedback was given in 100%, 0%, 25%, and 50% of the trials. Previous research reported by Ashby, Queller, and Berretty (1999) indicated that in an information-integration task, perfect accuracy was obtained supervised (100%) but not unsupervised (0%). Our results show that in both the 100% and 50% conditions, participants were able to achieve maximum accuracy. However, in the 0% and the 25% conditions, participants failed to learn. To discover the influence of the no-feedback trials on the learning process, the 50% condition was replicated in Experiment 2, substituting unrelated filler trials for the no feedback trials. The results indicated that accuracy rates were similar, suggesting no impact of the no-feedback trials on the learning process. The possibility of ever learning in a 25% setting was also researched in Experiment 2. Using twice as many trials, the results showed that all but 2 participants succeeded, suggesting that only the total number of feedback trials is important. The impact of the semisupervised learning results for ALCOVE, COVIS, and SPEED models is discussed. PMID- 19304623 TI - Implicit learning modulates selective attention at sensory levels of perceptual processing. AB - Electrophysiological evidence suggests that attention can be modulated as early as 100 msec after stimulus presentation. However, it is not clear whether these changes are based primarily on stimulus properties such as perceptual load (i.e., the level of perceptual difficulty), or other properties, such as general attentional set or learned expectations concerning perceptual load. Using event related potentials, this study examined how implicit learning of perceptual load conditions modulates selective attention at sensory levels of perceptual analysis. The results show significant differences in P1 amplitude recorded over occipital areas of the brain as a function of learned expectations of perceptual load, only when perceptual load could be reliably predicted by the preceding stimuli. Moreover, differences in processing were found when both low and high perceptual load conditions could be predicted. These findings suggest that implicit learning modulates the allocation of attention at early stages of perceptual processing. PMID- 19304624 TI - Auditory event files: integrating auditory perception and action planning. AB - The features of perceived objects are processed in distinct neural pathways, which call for mechanisms that integrate the distributed information into coherent representations (the binding problem). Recent studies of sequential effects have demonstrated feature binding not only in perception, but also across (visual) perception and action planning. We investigated whether comparable effects can be obtained in and across auditory perception and action. The results from two experiments revealed effects indicative of spontaneous integration of auditory features (pitch and loudness, pitch and location), as well as evidence for audio-manual stimulus-response integration. Even though integration takes place spontaneously, features related to task-relevant stimulus or response dimensions are more likely to be integrated. Moreover, integration seems to follow a temporal overlap principle, with features coded close in time being more likely to be bound together. Taken altogether, the findings are consistent with the idea of episodic event files integrating perception and action plans. PMID- 19304625 TI - Symbolic control of visual attention: semantic constraints on the spatial distribution of attention. AB - Humans routinely use spatial language to control the spatial distribution of attention. In so doing, spatial information may be communicated from one individual to another across opposing frames of reference, which in turn can lead to inconsistent mappings between symbols and directions (or locations). These inconsistencies may have important implications for the symbolic control of attention because they can be translated into differences in cue validity, a manipulation that is known to influence the focus of attention. This differential validity hypothesis was tested in Experiment 1 by comparing spatial word cues that were predicted to have high learned spatial validity ("above/below") and low learned spatial validity ("left/right"). Consistent with this prediction, when two measures of selective attention were used, the results indicated that attention was less focused in response to "left/right" cues than in response to "above/below" cues, even when the actual validity of each of the cues was equal. In addition, Experiment 2 predicted that spatial words such as "left/right" would have lower spatial validity than would other directional symbols that specify direction along the horizontal axis, such as "<--/-->" cues. The results were also consistent with this hypothesis. Altogether, the present findings demonstrate important semantic-based constraints on the spatial distribution of attention. PMID- 19304626 TI - Computation of mean size is based on perceived size. AB - The present study investigated whether computation of mean object size was based on perceived or physical size. The Ebbinghaus illusion was used to make the perceived size of a circle different from its physical size. Four Ebbinghaus configurations were presented either simultaneously (Experiment 1) or sequentially (Experiment 2) to each visual field, and participants were instructed to attend only to the central circles of each configuration. Participants' judgments of mean central circle size were influenced by the Ebbinghaus illusion. In addition, the Ebbinghaus illusion influenced the coding of individual size rather than the averaging. These results suggest that perceived rather than physical size was used in computing the mean size. PMID- 19304627 TI - Symbolic control of attention: tracking its temporal dynamics. AB - Three experiments examined the temporal dynamics of the impact of symbols with task-irrelevant spatial meanings on attentional control. In Experiments 1 and 2, participants were color-cued to report the first letter they saw in the left or right of two parallel letter streams. The cue appeared in the shape of an arrow pointing to the target stream (compatible) or to the other stream (incompatible). Incompatible arrows delayed letter selection; that is, participants reported later-appearing letters and made more errors. In Experiment 3, the target stream was cued in advance, and yet, incompatible symbols delayed target selection. These findings suggest that the irrelevant meaning of symbolic stimuli can still penetrate and bias attentional top-down control, an observation that challenges available control theories. PMID- 19304628 TI - The impact of multiple irrelevant visual events at the same spatial location on inhibition. AB - If an irrelevant visual event, such as a nonpredictive cue, is presented prior to a target, performance is impaired when the target appears at the cued location relative to when it is presented at an uncued location. This phenomenon, referred to as inhibition of return, can be found at multiple spatial locations when each is cued in succession. The present study examined the effect of successively cuing the same spatial location. Results suggested that additional inhibition occurred when more than one cue appeared at a single location at longer intercue intervals, but not at shorter intervals. These findings suggest that total inhibition to respond to targets at a spatial location reflects a summation of facilitatory and inhibitory factors generated by the presentation of each cue. PMID- 19304629 TI - Interocular suppression differentially affects achromatic and chromatic mechanisms. AB - Results from a series of psychophysical experiments show that interocular suppression produced by continuous flash suppression (CFS) differentially affects visual features of a target viewed by the other eye. When CFS stimuli are defined by luminance contrast, target color can be reliably identified but percent correct discrimination of target orientation is near chance. When the colored target is moving, color identification deteriorates with motion speed but direction of motion discrimination improves with target speed. Color's immunity to suppression is also weakened when interocular suppression is induced by equiluminant CFS stimuli that presumably stimulate the chromatic pathway. These results are discussed in terms of functional segregation of achromatic and chromatic processing in the visual system. PMID- 19304630 TI - The perception of subjective contours and neon color spreading figures in young infants. AB - The goal of the present habituation-dishabituation study was to explore sensitivity to subjective contours and neon color spreading patterns in infants. The first experiment was a replication of earlier investigations that showed evidence that even young infants are capable of perceiving subjective contours. Participants 4 months of age were habituated to a subjective Kanizsa square and were tested afterward for their ability to differentiate between the subjective square and a nonsubjective pattern that was constructed by rotating some of the inducing elements. Data analysis indicated a significant preference for the nonsubjective pattern. A control condition ensured that this result was not generated by the difference in figural symmetry or by the local differences between the test displays. In the second experiment, infant perception of a neon color spreading display was analyzed. Again, 4-month-old infants could discriminate between the illusory figure and a nonillusory pattern. Furthermore, infants in a control group did not respond to the difference in symmetry and the local differences between two nonillusory targets. Overall, the results show that young infants respond to illusory figures that are generated by either implicit T junctions (Experiment 1) or implicit X-junctions (Experiment 2). The findings are interpreted against the background of the neurophysiological model proposed by Grossberg and Mingolla (1985). PMID- 19304631 TI - Salient features in 3-D haptic shape perception. AB - Shape is an important cue for recognizing an object by touch. Several features, such as edges, curvature, surface area, and aspect ratio, are associated with 3-D shape. To investigate the saliency of 3-D shape features, we developed a haptic search task. The target and distractor items consisted of shapes (cube, sphere, tetrahedron, cylinder, and ellipsoid) that differed in several of these features. Exploratory movements were left as unconstrained as possible. Our results show that this type of haptic search task can be performed very efficiently (25 msec/item) and that edges and vertices are the most salient features. Furthermore, very salient local features, such as edges, can also be perceived through enclosure, an exploratory procedure usually associated with global shape. Since the subjects had to answer as quickly as possible, this suggests that speed may be a factor in selecting the appropriate exploratory procedure. PMID- 19304634 TI - The history of dipper functions. AB - Dipper-shaped curves often accurately depict the relationship between a baseline, or "pedestal," magnitude and a just noticeable difference in it. This tutorial traces the 45-year history of the dipper function in auditory and visual psychophysics, focusing on when they happen and why. Popular theories of both positive and negative masking (i.e., the "handle" and "dip," respectively) are described. Sometimes, but not always, negative masking disappears with an appropriate redescription of stimulus magnitude. PMID- 19304635 TI - Stimulus exposure and gaze bias: a further test of the gaze cascade model. AB - We tested predictions derived from the gaze cascade model of preference decision making (Shimojo, Simion, Shimojo, & Scheier, 2003; Simion & Shimojo, 2006, 2007). In each trial, participants' eye movements were monitored while they performed an eight-alternative decision task in which four of the items in the array were preexposed prior to the trial. Replicating previous findings, we found a gaze bias toward the chosen item prior to the response. However, contrary to the prediction of the gaze cascade model, preexposure of stimuli decreased, rather than increased, the magnitude of the gaze bias in preference decisions. Furthermore, unlike the prediction of the model, preexposure did not affect the likelihood of an item being chosen, and the pattern of looking behavior in preference decisions and on a non preference control task was remarkably similar. Implications of the present findings in multistage models of decision making are discussed. PMID- 19304636 TI - Knowledge alters visual contrast sensitivity. AB - Research has shown that the visual system's sensitivity to variations in luminance (visual contrast) within a particular area of the retina is affected in a bottom-up fashion by the ambient contrast levels in nearby regions. Specifically, changes in the ambient contrast in areas surrounding the target area alter the sensitivity to visual contrast within the target area. More recent research has shown that paying attention to the target or target area modulates contrast sensitivity, suggesting a top-down influence over contrast sensitivity that is mediated by attention. Here we report another form of top-down influence over contrast sensitivity that is unlikely to be mediated by attention. In particular, we show that knowledge and/or expectations about the levels of visual contrast that may appear in upcoming targets also affect how sensitive the observer is to the contrast in the target. This sort of knowledge-driven, top down contrast sensitivity control could be used to preset the visual system's contrast sensitivity to maximize discriminability and to protect contrast sensitive processes from a contrast overload. Overall, our results suggest that existing models of contrast sensitivity might benefit from the inclusion of top down control mechanisms. PMID- 19304637 TI - Lightness constancy: ratio invariance and luminance profile. AB - The term simultaneous lightness constancy describes the capacity of the visual system to perceive equal reflecting surfaces as having the same lightness despite lying in different illumination fields. In some cases, however, a lightness constancy failure occurs; that is, equal reflecting surfaces appear different in lightness when differently illuminated. An open question is whether the luminance profile of the illumination edges affects simultaneous lightness constancy even when the ratio invariance property of the illumination edges is preserved. To explore this issue, we ran two experiments by using bipartite illumination displays. Both the luminance profile of an illumination edge and the luminance ratio amplitude between the illumination fields were manipulated. Results revealed that the simultaneous lightness constancy increases when the luminance profile of the illumination edge is gradual (rather than sharp) and homogeneous (rather than inhomogeneous), whereas it decreases when the luminance ratio between the illumination fields is enlarged. The results are interpreted according to the layer decomposition schema, stating that the visual system splits the luminance into perceived lightness and apparent illumination components. We suggest that illumination edges having gradual and homogeneous luminance profiles facilitate the luminance decomposition process, whereas wide luminance ratios impede it. PMID- 19304638 TI - "2 x 3" primes naming "6": evidence from masked priming. AB - It is a common assumption for multiplication-solving models that single-digit multiplications are automatically retrieved. However, the experimental evidence for this is based on paradigms under suspicion. In this research, we employed a new procedure with the aim of assessing the automatic retrieval of multiplication more directly. In two experiments, multiplication automatism was studied using briefly presented primes (stimulus onset asynchrony = 48 msec) in a number-naming task. In Experiment 1, in the congruent conditions, the target and the prime were the same numbers (e.g., prime, 6; target, 6) or the target was the solution to the multiplication prime (e.g., prime, 2 x 3 = ; target, 6). In the incongruent conditions, no relationship existed between the primes and the targets (e.g., prime, 32; target, 6; or prime, 4 x 8 = ; target, 6). Experiment 2 explored the relevance of the equal sign for the multiplication-priming effect. Data showed that naming was faster when the solution of the multiplication prime matched the target, as compared with the incongruent condition (multiplication-priming effect), and that these effects were found irrespective of the presence of the equal sign. The fact that this priming effect was found even though the participants were unaware of the presentation of the primes supports the automatic character of single-digit multiplication. We conclude by arguing that this procedure is highly valuable for exploring the mechanisms involved in simple arithmetic solving. PMID- 19304639 TI - Tactile perception of thermal diffusivity. AB - The thermal diffusivity of an object is a parameter that controls the rate at which heat is extracted from the hand when it touches that object. It is an important feature for distinguishing materials by means of touch. In order to quantitatively describe the ability of human observers to discriminate between materials on the basis of heat extraction rate, we conducted an experiment in which this heat extraction was performed in a controlled way. In different conditions, subjects were repeatedly asked to select from two stimuli the one that cooled faster. The discrimination threshold was around 43% of the extraction rate. A rate that was twice as slow also yielded twice the absolute threshold. When we halved the temperature difference between the beginning and end of the stimulus, the threshold did not change as much. In separate experiments, we investigated the different cues that were available in the stimulus: initial cooling rate and end temperature. Both cues were used for discrimination, but cooling rate seemed to be the most important. PMID- 19304640 TI - Integration of multiple views of scenes. AB - In two experiments, memory was tested for changes in viewpoints in naturalistic scenes. In the key study condition, participants viewed two images of the same scene from viewpoints 40 degrees apart. There were two other study conditions: The two study images were identical or were of different scenes. A test image followed immediately, and participants judged whether it was identical to either of the study images. The scene in the test image was always the same as in a study image and was at least 20 degrees from any study image on different trials. Two models were tested: (1) views stored and retrieved independently and (2) views combined at retrieval. The crucial test of these hypotheses involved a comparison (in the key study condition) of the interpolation condition (the test image was presented between the two study images and 20 degrees from both) and the extrapolation condition (it was 20 degrees from one study image and 60 degrees from the other). Performance in the interpolation condition was far worse than what was predicted by the first model, whereas the second model fit the data quite well. The latter model is parsimonious in that it integrates previous experiences without requiring the integration of the views in memory. We review some of this model's broader implications. PMID- 19304641 TI - On the minimization of task switch costs following long-term training. AB - Flexible, context-dependent linkages between stimulus and response are fundamental to adaptive behavior. In the present article, we evaluate the limits of this flexibility by exploring the asymptotic efficiency of people's ability to switch between two different sensorimotor mappings. Two stimulus-response (S-R) mappings were learned, either both on the same hand (unimanual condition) or one mapping per hand (intermanual condition). The S-R mappings presented bivalent stimuli and employed the same response keys. A novel training regimen successfully reduced task-switch costs to approximately 20 msec, suggesting that residual switch costs cannot be eliminated. These costs cannot be entirely attributed to the cognitive control process of task-set reconfiguration, because they are observed over cued switch intervals of several seconds. Two additional issues in motor learning were addressed: the single or dual loci of manual motor control and the coordinate system of task representation. First, the results favored the notion of independent controllers for each hand instead of a single dominant controller, since intermanual performance was superior to unimanual performance. Second, a transfer task tested internal (egocentric) and external (allocentric) coordinate systems. Transfer was more effective using the external coordinate system, suggesting that the S-R mappings reflected the association between the bivalent stimuli and external goals (i.e., the individual keys) rather than the concrete pattern of muscle contractions (i.e., the finger pressing the key). Finally, retention tests revealed that these learned S-R associations were remarkably durable, since no decrements in fluent task switching performance were observed after 10 months without practice. PMID- 19304642 TI - Optic variables used to judge future ball arrival position in expert and novice soccer players. AB - Although many studies have looked at the perceptual-cognitive strategies used to make anticipatory judgments in sport, few have examined the informational invariants that our visual system may be attuned to. Using immersive interactive virtual reality to simulate the aerodynamics of the trajectory of a ball with and without sidespin, the present study examined the ability of expert and novice soccer players to make judgments about the ball's future arrival position. An analysis of their judgment responses showed how participants were strongly influenced by the ball's trajectory. The changes in trajectory caused by sidespin led to erroneous predictions about the ball's future arrival position. An analysis of potential informational variables that could explain these results points to the use of a first-order compound variable combining optical expansion and optical displacement. PMID- 19304643 TI - Amelioration of axis-aligned motion bias for active versus stationary judgments of bilaterally symmetric moving shapes' final destinations. AB - Recent research confirms that observers' judgments of projected final destinations of axis-trajectory misaligned moving figures are biased in the direction of primary axis deviation from trajectory, a phenomenon we named the axis-aligned motion (AAM) bias. The present study tests whether this bias occurs in a large, immersive mixed-reality environment that enables active (mobile) responses in making judgments of shapes' destinations. Like Morikawa (1999), we found that accuracy depended on axis-trajectory alignment and that there was a correspondence between final destination judgment error and the direction of axial deviation from the trajectory. Extending prior work, we found that comobile judgments were significantly more accurate than stationary ones for symmetric moving shapes, regardless of axial deviation, but only marginally so for asymmetric shapes. We conclude that our findings are ecologically consistent and that AAM is a natural regularity for which people have acquired a complementary perceptual-cognitive attunement: the AAM bias. PMID- 19304644 TI - Context influences holistic processing of nonface objects in the composite task. AB - We explored whether holistic-like effects can be observed for nonface objects in novices as a result of the task context. We measured contextually induced congruency effects for novel objects (Greebles) in a sequential matching selective attention task (composite task). When format at study was blocked, congruency effects were observed for study-misaligned, but not study-aligned, conditions (Experiment 1). However, congruency effects were observed in all conditions when study formats were randomized (Experiment 2), revealing that the presence of certain trial types (study-misaligned) in an experiment can induce congruency effects. In a dual task, a congruency effect for Greebles was induced in trials in which a face was first encoded, but only when it was aligned (Experiment 3). Thus, congruency effects can be induced by context that operates at the scale of the entire experiment or within a single trial. Implications for using the composite task to measure holistic processing are discussed. PMID- 19304645 TI - Even in correctable search, some types of rare targets are frequently missed. AB - Socially important visual search tasks, such as airport baggage screening and tumor detection, place observers in situations where the targets are rare and the consequences of failed detection are substantial. Recent laboratory studies have demonstrated that low target prevalence yields substantially higher miss errors than do high-prevalence conditions, in which the same targets appear frequently (Wolfe, Horowitz, & Kenner, 2005; Wolfe et al., 2007). Under some circumstances, this "prevalence effect" can be eliminated simply by allowing observers to correct their last response (Fleck & Mitroff, 2007). However, in three experiments involving search of realistic X-ray luggage images, we found that the prevalence effect is eliminated neither by giving observers the choice to correct a previous response nor by requiring observers to confirm their responses. This prevalence effect, obtained when no trial-by-trial feedback was given, was smaller than the effect obtained when observers searched through the same stimuli but were given trial-by-trial feedback about accuracy. We suggest that low prevalence puts pressure on observers in any search task, and that the diverse symptoms of that pressure manifest themselves differently in different situations. In some relatively simple search tasks, misses may result from motor or response errors. In other, more complex tasks, shifts in decision criteria appear to be an important contributor. PMID- 19304646 TI - A new look at novelty effects: guiding search away from old distractors. AB - We examined whether search is guided to novel distractors. In Experiment 1, subjects searched for a target among one new and a variable number of old distractors. Search displays in Experiment 2 consisted of an equal number of new, old, and familiar distractors (the latter repeated occasionally). We found that eye movements were preferentially directed to a new distractor on target-absent trials and that subjects tended to immediately fixate a new distractor after leaving the target on target-present trials. In both cases, first fixations on old distractors were consistently less frequent than could be explained by chance. We interpret these patterns as evidence for negative guidance: Subjects learn the visual features associated with the set of old distractors and then guide their search away from these features, ultimately resulting in the preferential fixation of novel distractors. PMID- 19304647 TI - Focused spatial attention is independent of rapid resumption of an interrupted search. AB - In three experiments, we examined possible relationships between the spatial focus of attention and the rapid resumption of a visual search following a brief interruption. In Experiment 1, we tested the role of involuntary (exogenous) spatial orienting to one region (quadrant) of a search display; in Experiment 2, we tested the role of voluntary (endogenous) spatial orienting to the same region; and in Experiment 3, we tested the role of voluntary orienting to the specific location in which the target item appeared. All three experiments indicated that spatial orienting speeds correct responding and greatly increases the probability of search success in the look immediately following the presentation of a spatial cue. However, these benefits of spatial cues were also shown to be completely independent of the rapid resumption effect, which depends on observers' forming a perceptual hypothesis about a target in one look, but being unable to confirm that hypothesis until a second one (Lleras, Rensink, & Enns, 2005). PMID- 19304648 TI - Illusory temporal order for stimuli at different depth positions. AB - We used four experiments to examine how the perceived temporal order of two visual stimuli depends on the depth position of the stimuli specified by a binocular disparity cue. When two stimuli were presented simultaneously at different depth positions in front of or around a fixation point, the observer perceived the more distant stimulus before the nearer stimulus (Experiments 1 and 2). This illusory temporal order was found only for sudden stimulus presentation (Experiment 3). These results suggest that a common processing, which is triggered by sudden luminance change, underlies this illusion. The strength of the illusion increased with the disparity gradient and the disparity size (Experiment 4). We propose that this illusion has a basis in the processing of motion in depth, which would alert the observer to a potential collision with an object that suddenly emerges in front of the observer. PMID- 19304649 TI - Processing of natural images is feedforward: a simple behavioral test. AB - Natural images can be classified so rapidly that it has been suggested that their analysis is based on a first single pass of processing activity through the visuomotor system. We tested this theory in a visuomotor priming task in which speeded pointing responses were performed toward one of two target images containing a prespecified stimulus (e.g., animal vs. non animal, ellipse vs. rectangle). Target pictures were preceded by prime pictures of the same or an opposite category, linked to either the same or an opposite pointing response. We found that pointing trajectories were initially controlled by the primes alone, but independently of information in the actual targets. Our data indicate that prime and target signals remained strictly sequential throughout all processing stages, meeting unprecedentedly stringent behavioral criteria for feedforward processing (rapid-chase criteria). Our findings suggest that visuomotor priming effects capture the output of the very first pass of information through the visuomotor system, before output is affected by recurrent information. PMID- 19304650 TI - The importance of semantics in auditory representations. AB - The purpose of the present study was to examine the nature of auditory representations by manipulating the semantic and physical relationships between auditory objects. On each trial, listeners heard a group of four simultaneous sounds for 1 sec, followed by 350 msec of noise, and then either the same sounds or three of the same plus a new one. Listeners completed a change-detection task and an object-encoding task. For change detection, listeners made a same different judgment for the two groups of sounds. Object encoding was measured by presenting probe sounds that either were or were not present in the two groups. In Experiments 1 and 3, changing the target to an object that was acoustically different from but semantically the same as the original target resulted in more errors on both tasks than when the target changed to an acoustically and semantically different object. In Experiment 2, comparison of semantic and acoustic effects demonstrated that acoustics provide a weaker cue than semantics for both change detection and object encoding. The results suggest that listeners rely more on semantic information than on physical detail.). PMID- 19304651 TI - Inhibition of saccadic eye movements to locations in spatial working memory. AB - Inhibition of return (IOR) refers to a bias against overt and covert attentional orienting toward previously attended locations. According to the reorienting hypothesis, IOR is generated when attention is withdrawn from the attended location and is prevented from "returning" to it. The present study investigated whether maintenance of attention at the cued location could affect the inhibition of oculomotor orienting to it. To preclude disengagement of attention, we asked participants to maintain the cued location in working memory. Maintenance of visuospatial information in memory has been shown to be accomplished through a sustained shift of spatial attention to a memorized location. Our results show that oculomotor IOR occurs at a particular location even when that location is kept in working memory (Experiment 1). Furthermore, we demonstrate that the mere act of maintenance of a location in working memory produces oculomotor inhibition similar to IOR (Experiments 2 and 3). We conclude that the oculomotor system is used for coding and maintaining locations in spatial working memory. In addition, we demonstrate that endogenous attention associated with maintenance of a location in working memory can be dissociated from the attention needed for execution of a saccadic eye movement. PMID- 19304652 TI - Color appearance: properties of the uniform appearance diagram derived from hue and saturation scaling. AB - Color appearance can be specified by a procedure of direct hue and saturation scaling. The scaling data can be represented on a 2-D color space termed a uniform appearance diagram (UAD). The orthogonal and bipolar axes of the UAD are based on the four unique hue sensations, which are blue-yellow and green-red. We have previously shown that the technique is reliable and rapid. We now show that the UAD is sufficiently uniform metrically that it can be used to derive, from a single set of scaling data, a wide range of color functions, such as the spectral loci of the unique hues, wavelength discrimination, and similarities among very different colored stimuli. The advantage of deriving a UAD is that it requires only a modest amount of participant time to generate the relevant data, which can be re-generated quickly to meet changing viewing conditions. PMID- 19304653 TI - Development of size constancy in children: a test of the metacognitive theory. AB - Two studies investigated children's abilities to estimate the sizes of distant objects. Each included a size estimation task and a size-distance knowledge test, which assessed children's understanding of the effects of distance on objects' image sizes and perceived sizes. In Study 1 (N = 79, age range = 5-10 years), high-knowledge children (those with above-median size-distance knowledge scores) made nearly accurate size estimates from a distance of 61 m and often reported using deliberate strategies to estimate size, whereas low-knowledge children underestimated size at this distance and typically reported no strategy use. In Study 2 (N = 60, age range = 6-11 years), high-knowledge children made nearly accurate size estimates from 61 m when given objective-size instructions and underestimated size when given apparent-size instructions. Low-knowledge children underestimated size in response to both instruction sets. The results suggest that age-related changes in size estimation accuracy result from the development of cognitive abilities necessary for using deliberate strategies to supplement perception. PMID- 19304654 TI - The danger of trial-by-trial knowledge of results in perceptual averaging studies. AB - Revived interest in "intuitive statistics" (Peterson & Beach, 1967) is evident in recent studies concerning the ability of observers to estimate mean size for ensembles of lines or of circles. To put the recent studies in context, and to highlight a potential danger in providing trial-by-trial knowledge of results (KOR), brief contact with previous research is made and a new experiment is presented demonstrating the malleability of responding to KOR. Together, these suggest two perils of KOR. First, given that the nature and even the existence of a molar size-averaging operator are both controversial, the proper criterion average for determining KOR is unclear. Second, whatever the operation observers use in this task, its scope and algorithm need to emerge from data unbiased by KOR. A corollary scaling concern is noted for averaging of areal stimuli (perceived as a compression function of physical area) as an instance of the more general concern that perceptual scaling factors must be taken into account when psychological averaging processes are studied. PMID- 19304655 TI - Small margin excision of periocular basal cell carcinomas. AB - AIMS: To analyse the outcome of small margin (up to 2 mm) excision of primary clinically well-defined periocular basal cell carcinomas (BCCs). METHODS: Retrospective evaluation of 90 well-demarcated primary BCCs having a minimum follow-up of 36 months. All patients underwent excision of the tumour with maximum margins of 2 mm. Resulting defects were, if possible, closed directly. Reconstruction of defects requiring flaps or grafts was delayed until receipt of the histological report which was obtained in all cases. RESULTS: One-stage excision and direct closure was performed in 67 patients (74.4%). In 23 patients (25.6%) reconstruction was delayed by 4 days to enable receipt of the histopathology report. Histological assessment confirmed complete excision after the first excision in 78 (86.7%) rising to 83 (92.2%) after two excisions. The mean follow-up was 47.5 (SD 9.1) months. Of the 12 cases with incompletely excised lesions, seven of the patients chose not to have any more surgery, and only one of these recurred. There were two other recurrences, and in both of them the lesion had initially been reported as completely excised. Overall the recurrence rate in our study was 3.3% (n = 3). CONCLUSIONS: Our recurrence rate compares well with published results following conventional excision of BCCs. In the absence of availability of Mohs surgery, well-demarcated nodular basal cell carcinomas can be safely excised using smaller margins than conventionally practised. PMID- 19304656 TI - Analysis of assembly and trafficking of native P2X4 and P2X7 receptor complexes in rodent immune cells. AB - P2X4 and P2X7 are the predominant P2X receptor subtypes expressed in immune cells. Having previously shown a structural and functional interaction between the two recombinant receptors, our aims here were to identify the preferred assembly pathway of the endogenous receptors in macrophage-like cells and to investigate the trafficking of these receptors between the plasma membrane and intracellular sites. We exploited the difference in size between the two subunits, and we used a combination of cross-linkers and blue native-PAGE analysis to investigate the subunit composition of complexes present in primary cultures of rat microglia and macrophages from wild type and P2X7(-/-) mice. Our results indicate that the preferred assembly pathway for both receptors is the formation of homotrimers. Homotrimers of P2X7 were able to co-immunoprecipitate with P2X4, suggesting that an interaction occurs between rather than within receptor complexes. In both macrophages and microglia, P2X7 receptors were predominantly at the cell surface, whereas P2X4 receptors were predominantly intracellular. There were clear cell type-dependent differences in the extent to which P2X4 receptors trafficked to and from the surface; trafficking was much more dynamic in microglia than in the macrophages, and further activation of cultured microglia with relatively short (3-h) incubations with lipopolysaccharide caused an approximately 4-fold increase in the fraction of receptors at the surface with only a 1.2-fold increase in total expression. The redistribution of intracellular receptors is thus an efficient means of enhancing the functional expression of P2X4 at the plasma membrane of microglia. PMID- 19304657 TI - The armadillo repeat-containing protein, ARMCX3, physically and functionally interacts with the developmental regulatory factor Sox10. AB - Sox10 is a member of the group E Sox transcription factor family and plays key roles in neural crest development and subsequent cellular differentiation. Sox10 binds to regulatory sequences in target genes via its conserved high mobility group domain. In most cases, Sox10 exerts its transcriptional effects in concert with other DNA-binding factors, adaptor proteins, and nuclear import proteins. These interactions can lead to synergistic gene activation and can be cell type specific. In earlier work, we demonstrated that Sox10 transactivates the nicotinic acetylcholine receptor alpha3 and beta4 subunit genes and does so only in neuronal-like cell lines, raising the possibility that Sox10 mediates its effects via interactions with co-regulatory factors. Here we describe the identification of the armadillo repeat-containing protein, ARMCX3, as a Sox10 interacting protein. Biochemical analyses indicate that ARMCX3 is an integral membrane protein of the mitochondrial outer membrane. Others have shown that Sox10 is a nucleocytoplasmic shuttling protein. We extend this observation and demonstrate that, in the cytoplasm, Sox10 is peripherally associated with the mitochondrial outer membrane. Both Sox10 and ARMCX3 are expressed in mouse brain and spinal cord as well as several cell lines. Overexpression of ARMCX3 increased the amount of mitochondrially associated Sox10. In addition, although ARMCX3 does not possess intrinsic transcriptional activity, it does enhance transactivation of the nicotinic acetylcholine receptor alpha3 and beta4 subunit gene promoters by Sox10. These results suggest that Sox10 is a membrane-associated factor whose transcriptional function is increased by direct interactions with ARMCX3 and raise the possibility of a signal transduction cascade between the nucleus and mitochondria through Sox10/ARMCX3 interactions. PMID- 19304658 TI - Involvement of all-trans-retinal in acute light-induced retinopathy of mice. AB - Exposure to bright light can cause visual dysfunction and retinal photoreceptor damage in humans and experimental animals, but the mechanism(s) remain unclear. We investigated whether the retinoid cycle (i.e. the series of biochemical reactions required for vision through continuous generation of 11-cis-retinal and clearance of all-trans-retinal, respectively) might be involved. Previously, we reported that mice lacking two enzymes responsible for clearing all-trans retinal, namely photoreceptor-specific ABCA4 (ATP-binding cassette transporter 4) and RDH8 (retinol dehydrogenase 8), manifested retinal abnormalities exacerbated by light and associated with accumulation of diretinoid-pyridinium-ethanolamine (A2E), a condensation product of all-trans-retinal and a surrogate marker for toxic retinoids. Now we show that these mice develop an acute, light-induced retinopathy. However, cross-breeding these animals with lecithin:retinol acyltransferase knock-out mice lacking retinoids within the eye produced progeny that did not exhibit such light-induced retinopathy until gavaged with the artificial chromophore, 9-cis-retinal. No significant ocular accumulation of A2E occurred under these conditions. These results indicate that this acute light induced retinopathy requires the presence of free all-trans-retinal and not, as generally believed, A2E or other retinoid condensation products. Evidence is presented that the mechanism of toxicity may include plasma membrane permeability and mitochondrial poisoning that lead to caspase activation and mitochondria associated cell death. These findings further understanding of the mechanisms involved in light-induced retinal degeneration. PMID- 19304659 TI - Mechanism of sustained activation of ribosomal S6 kinase (RSK) and ERK by kaposi sarcoma-associated herpesvirus ORF45: multiprotein complexes retain active phosphorylated ERK AND RSK and protect them from dephosphorylation. AB - As obligate intracellular parasites, viruses exploit diverse cellular signaling machineries, including the mitogen-activated protein-kinase pathway, during their infections. We have demonstrated previously that the open reading frame 45 (ORF45) of Kaposi sarcoma-associated herpesvirus interacts with p90 ribosomal S6 kinases (RSKs) and strongly stimulates their kinase activities ( Kuang, E., Tang, Q., Maul, G. G., and Zhu, F. (2008) J. Virol. 82, 1838-1850 ). Here, we define the mechanism by which ORF45 activates RSKs. We demonstrated that binding of ORF45 to RSK increases the association of extracellular signal-regulated kinase (ERK) with RSK, such that ORF45, RSK, and ERK formed high molecular mass protein complexes. We further demonstrated that the complexes shielded active pERK and pRSK from dephosphorylation. As a result, the complex-associated RSK and ERK were activated and sustained at high levels. Finally, we provide evidence that this mechanism contributes to the sustained activation of ERK and RSK in Kaposi sarcoma-associated herpesvirus lytic replication. PMID- 19304660 TI - The activity of the epithelial sodium channels is regulated by caveolin-1 via a Nedd4-2-dependent mechanism. AB - It has recently been shown that the epithelial Na(+) channel (ENaC) is compartmentalized in caveolin-rich lipid rafts and that pharmacological depletion of membrane cholesterol, which disrupts lipid raft formation, decreases the activity of ENaC. Here we show, for the first time, that a signature protein of caveolae, caveolin-1 (Cav-1), down-regulates the activity and membrane surface expression of ENaC. Physical interaction between ENaC and Cav-1 was also confirmed in a coimmunoprecipitation assay. We found that the effect of Cav-1 on ENaC requires the activity of Nedd4-2, a ubiquitin protein ligase of the Nedd4 family, which is known to induce ubiquitination and internalization of ENaC. The effect of Cav-1 on ENaC requires the proline-rich motifs at the C termini of the beta- and gamma-subunits of ENaC, the binding motifs that mediate interaction with Nedd4-2. Taken together, our data suggest that Cav-1 inhibits the activity of ENaC by decreasing expression of ENaC at the cell membrane via a mechanism that involves the promotion of Nedd4-2-dependent internalization of the channel. PMID- 19304661 TI - The carboxyl-terminal domain of atypical protein kinase Czeta binds to ceramide and regulates junction formation in epithelial cells. AB - Atypical protein kinase Cs (PKCs) (aPKCzeta and lambda/iota) have emerged as important binding partners for ceramide, a membrane-resident cell signaling lipid that is involved in the regulation of apoptosis as well as cell polarity. Using ceramide overlay assays with proteolytic fragments of PKCzeta and vesicle binding assays with ectopically expressed protein, we show that a protein fragment comprising the carboxyl-terminal 20-kDa sequence of PKCzeta (C20zeta, amino acids 405-592) bound to C16:0 ceramide. This sequence is not identical to the C1 domain (amino acids 131-180), which has been suggested to serve as a potential ceramide binding domain. Using immunocytochemistry, we found that a C20zeta protein fragment ectopically expressed in two epithelial cell types (neural progenitors and Madin-Darby canine kidney cells) co-distributed with ceramide. Stable expression of C20zeta-EGFP in Madin-Darby canine kidney cells disrupted the formation of adherens and tight junctions and impaired the epithelium integrity by reducing transepithelial electrical resistance. Disruption of cell adhesion and loss of transepithelial electrical resistance was prevented by incubation with C16:0 ceramide. Our results show, for the first time, that there is a novel ceramide binding domain (C20zeta) in the carboxyl terminus of aPKC. Our results also show that the interaction of ceramide with this binding domain is essential for cell-to-cell contacts in epithelia. Therefore, ceramide interaction with the C20zeta binding domain is a potential mechanism by which ceramide and aPKC regulate the formation of junctional complexes in epithelial cells. PMID- 19304662 TI - Sexual stage adhesion proteins form multi-protein complexes in the malaria parasite Plasmodium falciparum. AB - The sexual phase of the malaria parasite Plasmodium falciparum is accompanied by the coordinated expression of stage-specific adhesive proteins. Among these are six secreted proteins with multiple adhesion domains, termed P. falciparum LCCL domain-containing protein (PfCCp) proteins, which are expressed in the parasitophorous vacuole of the differentiating gametocytes and which are later associated with macrogametes. Although the majority of the PfCCp proteins are implicated in parasite development in the mosquito vector, their functions remain unknown. In the present study we investigated the molecular interactions between the PfCCp proteins during gametocyte development and emergence. Using five different gene-disruptant parasite lines, we show that the lack of one PfCCp protein leads to the loss of other PfCCp family members. Co-immunoprecipitation assays on gametocyte lysates revealed formation of complexes involving all PfCCp proteins, and affinity chromatography co-elution binding assays with recombinant PfCCp domains further indicated direct binding between distinct adhesion domains. PfCCp-coated latex beads bind to newly formed macrogametes but not to gametocytes or older macrogametes 6 or 24 h post-activation. In view of these data, we propose that the PfCCp proteins form multi-protein complexes that are exposed during gametogenesis, thereby mediating cell contacts of macrogametes. PMID- 19304663 TI - Spectroscopic insights into the oxygen-tolerant membrane-associated [NiFe] hydrogenase of Ralstonia eutropha H16. AB - This study provides the first spectroscopic characterization of the membrane bound oxygen-tolerant [NiFe] hydrogenase (MBH) from Ralstonia eutropha H16 in its natural environment, the cytoplasmic membrane. The H2-converting MBH is composed of a large subunit, harboring the [NiFe] active site, and a small subunit, capable in coordinating one [3Fe4S] and two [4Fe4S] clusters. The hydrogenase dimer is electronically connected to a membrane-integral cytochrome b. EPR and Fourier transform infrared spectroscopy revealed a strong similarity of the MBH active site with known [NiFe] centers from strictly anaerobic hydrogenases. Most redox states characteristic for anaerobic [NiFe] hydrogenases were identified except for one remarkable difference. The formation of the oxygen-inhibited Niu-A state was never observed. Furthermore, EPR data showed the presence of an additional paramagnetic center at high redox potential (+290 mV), which couples magnetically to the [3Fe4S] center and indicates a structural and/or redox modification at or near the proximal [4Fe4S] cluster. Additionally, significant differences regarding the magnetic coupling between the Nia-C state and [4Fe4S] clusters were observed in the reduced form of the MBH. The spectroscopic properties are discussed with regard to the unusual oxygen tolerance of this hydrogenase and in comparison with those of the solubilized, dimeric form of the MBH. PMID- 19304664 TI - Familial FTDP-17 missense mutations inhibit microtubule assembly-promoting activity of tau by increasing phosphorylation at Ser202 in vitro. AB - In Alzheimer disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) and other tauopathies, tau accumulates and forms paired helical filaments (PHFs) in the brain. Tau isolated from PHFs is phosphorylated at a number of sites, migrates as approximately 60-, 64-, and 68-kDa bands on SDS gel, and does not promote microtubule assembly. Upon dephosphorylation, the PHF tau migrates as approximately 50-60-kDa bands on SDS-gels in a manner similar to tau that is isolated from normal brain and promotes microtubule assembly. The site(s) that inhibits microtubule assembly-promoting activity when phosphorylated in the diseased brain is not known. In this study, when tau was phosphorylated by Cdk5 in vitro, its mobility shifted from approximately 60-kDa bands to approximately 64- and 68-kDa bands in a time-dependent manner. This mobility shift correlated with phosphorylation at Ser(202), and Ser(202) phosphorylation inhibited tau microtubule-assembly promoting activity. When several tau point mutants were analyzed, G272V, P301L, V337M, and R406W mutations associated with FTDP-17, but not nonspecific mutations S214A and S262A, promoted Ser(202) phosphorylation and mobility shift to a approximately 68-kDa band. Furthermore, Ser(202) phosphorylation inhibited the microtubule assembly-promoting activity of FTDP-17 mutants more than of WT. Our data indicate that FTDP-17 missense mutations, by promoting phosphorylation at Ser(202), inhibit the microtubule assembly-promoting activity of tau in vitro, suggesting that Ser(202) phosphorylation plays a major role in the development of NFT pathology in AD and related tauopathies. PMID- 19304665 TI - Heterologous stacking of prion protein peptides reveals structural details of fibrils and facilitates complete inhibition of fibril growth. AB - Fibrils play an important role in the pathogenesis of amyloidosis; however, the underlying mechanisms of the growth process and the structural details of fibrils are poorly understood. Crucial in the fibril formation of prion proteins is the stacking of PrP monomers. We previously proposed that the structure of the prion protein fibril may be similar as a parallel left-handed beta-helix. The beta helix is composed of spiraling rungs of parallel beta-strands, and in the PrP model residues 105-143 of each PrP monomer can contribute two beta-helical rungs to the growing fibril. Here we report data to support this model. We show that two cyclized human PrP peptides corresponding to residues 105-124 and 125-143, based on two single rungs of the left-handed beta-helical core of the human PrP(Sc) fibril, show spontaneous cooperative fibril growth in vitro by heterologous stacking. Because the structural model must have predictive value, peptides were designed based on the structure rules of the left-handed beta helical fold that could stack with prion protein peptides to stimulate or to block fibril growth. The stimulator peptide was designed as an optimal left handed beta-helical fold that can serve as a template for fibril growth initiation. The inhibiting peptide was designed to bind to the exposed rung but frustrate the propagation of the fibril growth. The single inhibitory peptide hardly shows inhibition, but the combination of the inhibitory with the stimulatory peptide showed complete inhibition of the fibril growth of peptide huPrP-(106-126). Moreover, the unique strategy based on stimulatory and inhibitory peptides seems a powerful new approach to study amyloidogenic fibril structures in general and could prove useful for the development of therapeutics. PMID- 19304666 TI - Specific expression and regulation of hepassocin in the liver and down-regulation of the correlation of HNF1alpha with decreased levels of hepassocin in human hepatocellular carcinoma. AB - Hepassocin (HPS), is a liver-specific gene with mitogenic activity on isolated hepatocytes. It is up-regulated following partial hepatectomy and down-regulated frequently in heptocellular carcinoma (HCC). However, very little is known about the HPS transcription regulation mechanism. In this study, we identified HNF1alpha (hepatocyte nuclear factor-1alpha) as an important liver-specific cis acting element for HPS using in vivo luciferase assays. Deletion of the HNF1 binding site not only led to a complete loss of HPS promoter activity in vivo but also abolished the induction of the HPS promoter by HNF1alpha. An electrophoretic mobility shift assay demonstrated that HNF1alpha interacted with the HPS gene promoter in vitro. Chromatin immunoprecipitation showed that HNF1alpha interacted with HMGB1 and CREB-binding protein, and all of them were recruited to the HPS promoter in vivo. Moreover, HNF1alpha expression was lower in HCC cell lines and tissues and correlated significantly with the down-regulation of HPS expression. Re-expression of HNF1alpha in human hepatoma HepG2 cells reinduced HPS expression. In contrast, knockdown of endogenous HNF1alpha expression by small interfering RNA resulted in a significant reduction of HPS expression. Furthermore, we found that partial hepatectomy and IL-6 significantly induced promoter activity of HPS, depending on STAT3 and HNF1 binding sites in the HPS promoter. These results demonstrate that the HNF1 binding site and HNF1alpha are critical to liver-specific expression of HPS, and down-regulation or loss of HNF1alpha causes, at least in part, the transcriptional down-regulation of HPS in HCC. PMID- 19304667 TI - Relationships between alumina and bauxite dust exposure and cancer, respiratory and circulatory disease. AB - OBJECTIVES: To examine the associations between alumina and bauxite dust exposure and cancer incidence and circulatory and respiratory disease mortality among bauxite miners and alumina refinery workers. METHODS: This cohort of 5770 males has previously been linked to national mortality and national and state cancer incidence registries (1983-2002). In this paper, Poisson regression was used to undertake internal comparisons within the cohort based on subgroups of cumulative exposure to inhalable bauxite and alumina dust. Exposure was estimated using job histories and historical air monitoring data. RESULTS: There was no association between ever bauxite exposure and any of the outcomes. There was a borderline significant association between ever alumina exposure and cerebrovascular disease mortality (10 deaths, RR 3.8, 95% CI 1.1 to 13). There was some evidence of an exposure-response relationship between cumulative bauxite exposure and non malignant respiratory disease mortality (seven deaths, trend p value: 0.01) and between cumulative alumina exposure and cerebrovascular disease mortality (trend p value: 0.04). These associations were based on very few cases and for non malignant respiratory disease the deaths represented a heterogeneous mixture of causes. There was no evidence of an excess risk for any cancer type with bauxite or alumina exposure. CONCLUSIONS: These preliminary findings, based on very few cases, suggest that cumulative inhalable bauxite exposure may be associated with an excess risk of death from non-malignant respiratory disease and that cumulative inhalable alumina dust exposure may be associated with an excess risk of death from cerebrovascular disease. Neither exposure appears to increase the risk of incident cancers. PMID- 19304668 TI - Effect of cardiac rehabilitation on angiogenic cytokines in postinfarction patients. AB - OBJECTIVE: To determine whether cardiac rehabilitation influences plasma levels of angiogenic cytokines and their correlation with myocardial blood flow (MBF). DESIGN: Randomised controlled study. SETTING: Tertiary cardiac centre. PATIENTS: 39 postinfarction patients randomised to either a 3-month training group (n = 20) or a non-training group (n = 19), and 19 normal controls. INTERVENTIONS: Cardiac rehabilitation. MAIN OUTCOME MEASURES: MBF by cardiac magnetic resonance imaging, and plasma levels of stem cell factor (SCF), stromal-derived factor-1 (SDF-1), and vascular endothelial growth factor (VEGF) measured at enrolment and at 3 months after randomisation. RESULTS: At baseline, when compared with the healthy subjects, postinfarction patients had a lower MBF in the infarcted myocardium during dipyridamole-induced stress (1.65 (0.58) vs 2.77 (0.78) ml/min/g, p<0.001) but higher plasma levels of VEGF (3.65 (0.75) vs 2.77 (0.59) pg/ml, p<0.001 expressed as the natural logarithm) and SDF-1 (2113 (345) vs 1869 (309) pg/ml, p = 0.009). Only SDF-1 was inversely associated with stress MBF in both remote (r = -0.39, p = 0.03) and infarcted myocardium (r = -0.62, p<0.001). After 3 months, the training group's stress MBF had increased by 33% in the remote (p<0.001) and 28% in infarcted myocardium (p = 0.02), while VEGF decreased by 9% (p = 0.01), and SDF-1 decreased by 11% (p = 0.02). The change in SDF-1 was inversely correlated with the change in stress MBF in both remote (r = -0.40, p = 0.01) and infarcted myocardium (r = -0.50, p = 0.001). In the non-training group, MBF and cytokines were unchanged. CONCLUSION: Cardiac rehabilitation improves stress MBF in postinfarction patients, with an inverse decrease in circulating angiogenic cytokines. PMID- 19304669 TI - The prognosis of patients with atrial fibrillation is improved when sinus rhythm is restored: report from the Stockholm Cohort of Atrial Fibrillation (SCAF). AB - BACKGROUND: Clinical trials have indicated that an active rhythm control strategy aiming at restoration of sinus rhythm in patients with atrial fibrillation (AF) is no better than a rate-control strategy in terms of mortality and morbidity. To what extent restoration and maintenance of sinus rhythm per se affect long-term prognosis in AF patients is less clear. AIM: To investigate if there are differences in mortality and morbidity between direct current (DC)-cardioverted AF patients who remain in sinus rhythm after cardioversion and those who relapse early. METHOD: 361 cardioverted patients from the Stockholm Cohort Study on Atrial Fibrillation were studied by means of medical records and national registers. Patients were followed for a mean of 4.2 years from DC cardioversion regarding all-cause mortality and for a mean of 3.2 years for a composite endpoint of death, ischaemic stroke, myocardial infarction or hospitalisation for heart failure. RESULTS: All-cause mortality tended to be lower in patients who had been successfully cardioverted and had had no known relapse of AF within the first 3 months after cardioversion (hazard ratio (HR) 0.57, 95% CI 0.30 to 1.06, p = 0.076). They also had a significantly lower incidence of the composite endpoint than those who relapsed early (HR 0.51, 95% CI 0.32 to 0.82, p = 0.0058). CONCLUSION: Restoration and 3 months maintenance of sinus rhythm was associated with improved long-term prognosis. The results imply that an active DC cardioversion approach is justified. PMID- 19304670 TI - Prenatal diagnosis of left atrial isomerism. AB - OBJECTIVE: To describe the cardiac anomalies and outcome in the fetus with left atrial isomerism. METHODS: All fetuses with a diagnosis of left atrial isomerism between 1998 and 2008 were identified. Gestational age at diagnosis, the nuchal translucency, the karyotype, the cardiac findings and outcome were noted. A literature search from 1990 identified four publications reporting 10 or more cases of fetal left atrial isomerism. The same data, where available, were collected from these papers for comparison. RESULTS: There were 41 fetuses with this diagnosis seen in our centre. All cases had an interrupted inferior vena cava with azygous continuation. Associated cardiac defects were similar in our series and in the 129 cases reported in the literature and are therefore grouped together. They included complete atrioventricular septal defect (68%), complete heart block (38%), viscerocardiac heterotaxy (54%), double outlet right ventricle (23%), right ventricular outflow tract obstruction (35%), left ventricular outflow tract obstruction (21%) and total anomalous pulmonary vein drainage (5%). In our series, there were 22 pregnancy terminations, seven intrauterine deaths, one neonatal death, one infant death and one was lost to follow-up. Of the continuing pregnancies only 50% in our series and 60% in the reported series survived. CONCLUSION: Left atrial isomerism presents a varied spectrum of cardiac malformations when it is detected prenatally. Complete heart block, complex cardiac abnormalities and fetal hydrops are poor prognostic features. Those with only minor cardiac malformations are at risk postnatally for biliary atresia and for bowel obstruction due to malrotation. PMID- 19304671 TI - Sirolimus-eluting stents, bare metal stents or coronary artery bypass grafting for patients with multivessel disease including involvement of the proximal left anterior descending artery: analysis of the Arterial Revascularization Therapies study part 2 (ARTS-II). AB - OBJECTIVE: The The Arterial Revascularization Therapies Study (ARTS)-II trial found no differences in survival or overall adverse events between sirolimus eluting stents (SES) and the surgical arm of ARTS-I. Nevertheless, existing data suggest that patients with disease of the proximal left anterior descending artery (LAD) may derive particular benefit from coronary artery bypass grafting (CABG). We therefore analysed the clinical outcome of patients in ARTS-I and ARTS II with proximal LAD involvement. DESIGN: Multicentre observational study. SETTING: Forty-five European academic hospitals. PATIENTS: Patients with multivessel coronary artery disease. INTERVENTIONS: Patients in ARTS-II with proximal LAD disease treated with SES (289/607, 48%) were compared with 187/600 (31%) bare metal stent patients (ARTS-I BMS) and 206/605 (34%) surgical patients (ARTS-I CABG) with proximal LAD involvement from ARTS-I. MAIN OUTCOME MEASURES: Major adverse cardiac and cerebrovascular events after 3 years. RESULTS: The Arterial Revascularization Therapies study part 2 (ARTS-II) subgroup had better survival than both ARTS-I groups (ARTS-II 98.6% vs ARTS-I BMS 95.7%, p = 0.05 and vs ARTS-I CABG 94.7%, p = 0.01) and lower rates of the hard clinical composite endpoint of death or non-fatal myocardial infarction (ARTS-II 3.1% vs ARTS-I BMS 9.6%, p = 0.002 and vs ARTS-I CABG 9.7%, p = 0.002). Although the ARTS-I CABG patients had a lower need for repeat revascularisation than ARTS-II (5.3% vs 13.1%, p = 0.002), the overall composite adverse event rates (death, myocardial infarction, stroke or any repeat revascularisation) were not significantly different between the ARTS-I CABG and ARTS-II patients (15.0% vs 18.0%, p = 0.4). CONCLUSIONS: SES are not inferior to CABG or bare metal stents for the treatment of patients with multivessel coronary disease including involvement of the proximal LAD. PMID- 19304672 TI - Genetic influences on adult body mass index followed over 29 years and their effects on late-life mobility: a study of twin sisters. AB - BACKGROUND: The rise in body mass index (BMI) during adulthood increases the risk for metabolic disorders, functional limitations and disability in old age. This twin study examined prospectively whether genetic and environmental influences on women's BMI also account for mobility 29 years later. METHODS: The sample consisted of 103 monozygotic and 114 dizygotic pairs of twin sisters reared together. Body mass index was initially evaluated in 1975, when the women were aged 42.6+/-3.4 years, and was followed-up in 1981, 1990, 2001 and 2004. Mobility was evaluated using the standardised 6-minute walking test in 2001, when the women were aged 68.6+/-3.2 years, and followed-up 3 years later. An investigation was made into how genetic and environmental influences on adult BMI accounted for mobility in old age using a genetic latent growth modelling approach. RESULTS: During the follow-up period, BMI increased by 17%. Midlife BMI was a significant predictor of mobility 29 years later. Genetic influences on BMI level and its rate-of-change accounted for 37% and 25% respectively, of the genetic influences on mobility later in life. The corresponding environmental influences on BMI level and its rate-of-change were 35% and 22%. CONCLUSION: Genes predisposing to higher BMI across middle age increase the risk for poorer mobility in old age. Identifying those genes could lead to interventions targeted at preventing obesity and mobility loss later in life. However, modification of environmental factors, eg exercise and nutrition, remain the most feasible ways of influencing BMI and mobility across the life span. PMID- 19304673 TI - Inflammation imaging in atherosclerosis. AB - Inflammation is important at many stages of atherosclerotic plaque development. We highlight several imaging modalities that can quantify the degree of plaque inflammation noninvasively. Imaging of this type might allow testing of novel antiatherosclerosis drugs, identification of patients at risk of plaque rupture, and deeper insight into the biology of the disease. The imaging modalities are discussed in relation to their potential use in these areas. PMID- 19304674 TI - The usefulness of virtual MR arthroscopy as an adjunct to conventional MR arthrography in detecting anterior labral lesions of the shoulder. AB - OBJECTIVE: The objective of this study was to investigate the usefulness of virtual MR arthroscopy as an adjunct to conventional MR arthrography in detecting various anteroinferior glenoid labral lesions. MATERIALS AND METHODS: The study group was composed of 28 patients (22 men and six women; mean age, 25 years) who underwent direct MR arthrography of the shoulder as well as arthroscopy due to recurrent shoulder dislocations. MR arthrography examinations were reviewed retrospectively with radiologists blinded to the arthroscopic diagnoses. Virtual MR arthroscopy was produced from fat-suppressed 3D gradient-recalled echo MR arthrography using volume-rendering software. The sensitivity, specificity, and accuracy for diagnosis were calculated. The diagnostic performances were validated by receiver operating characteristic curve analysis. RESULTS: The structure of the glenoid labrum was well visualized in a 3D perspective, and characterization of the injured anterior glenoid labrum was aided by adjunct use of virtual MR arthroscopy. The accuracy for detecting classic Bankart, anterior labroligamentous periosteal sleeve avulsion, and Perthes lesions improved to 85.7%, 85.7%, and 92.9%, respectively, with adjunct virtual MR arthroscopy with MR arthrography versus 78.6%, 82.1%, and 89.3%, respectively, with MR arthrography alone. A statistically significant improvement of the diagnostic capability of MR arthrography with adjunct virtual MR arthroscopy was proven for the detection of a classic Bankart lesion (p < 0.05). CONCLUSION: Having a 3D perspective to validate chronic recurrent glenoid labral injuries of the shoulder facilitated understanding of the anatomic relationships and helped diagnosis. Adjunct virtual MR arthroscopy is potentially useful for detecting and characterizing chronic recurrent glenoid labral lesions to improve diagnosis. PMID- 19304675 TI - Prognostic value of 18F-FDG PET in patients with head and neck squamous cell cancer. AB - OBJECTIVE: This study was designed to assess whether tumor uptake of (18)F-FDG (FDG) expressed as the standardized uptake value (SUV) can be used to predict survival in patients with head and neck cancer. Furthermore, a prognostic maximum SUV was determined with univariate and bivariate analyses. CONCLUSION: Low SUVs ( 7.0). In the Cox proportional hazards model, tumor SUV was a significant and independent predictor of local control (p = 0.022) and disease-free survival (p = 0.019). In addition, in the group of high SUV, high T stage was more associated with poorer outcome than low T stage (p = 0.0502). Therefore, patients with higher tumor FDG uptake should be considered for a more aggressive treatment approach. PMID- 19304676 TI - Freehand real-time MRI-guided lumbar spinal injection procedures at 1.5 T: feasibility, accuracy, and safety. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that freehand real-time MRI-guided lumbar spinal injection procedures are feasible, accurate, and safe when performed with a clinical open-bore 1.5-T MRI system. MATERIALS AND METHODS: A retrospective analysis was made of spinal injection procedures performed over an 18-month period. Forty-nine procedures were performed on 37 patients (23 women, 14 men; mean age, 36 years; range, 18-48 years). A rapid FLASH 2D MRI sequence (TR/TE, 9.3/3.5; slice thickness, 5 mm; acquisition time, 1 second) was used for real-time needle placement with freehand technique. Data assessed were type of procedure, qualitative and quantitative image quality, dimensions of needle artifact, rate of successful drug delivery, rate of vascular uptake, time requirements, and occurrence of complications. RESULTS: Among the 49 procedures, 22 (45%) were nerve root injections, 18 (37%) were facet joint injections, and nine (18%) were epidural injections. The quality of real-time FLASH 2D MR images was sufficient in all cases. Contrast-to-noise ratios were sufficiently high for good delineation of relevant structures. The needle artifact made was remarkably constant with an average overestimation of length of 1.0 +/- 0.2 [SD] mm. Drug delivery was successful in all selective nerve root injections and epidural injections. The rate of successful drug delivery was 89% (16 of 18) for facet joint injections. No complete intravascular injections occurred. The mean table time was 36 minutes (range, 23-75 minutes). The mean real-time MRI time was 38 seconds (range, 12-185 seconds). No major complications occurred. CONCLUSION: We accept the hypothesis that freehand real-time MRI-guided lumbar spinal injection procedures are feasible, accurate, and safe when performed with a clinical open-bore 1.5-T MRI system. We note that real-time MRI guidance has the additional advantage of a complete absence of patient and operator exposure to ionizing radiation. PMID- 19304677 TI - A novel interventional radiology technique for arterial infusion chemotherapy against advanced pancreatic cancer. AB - OBJECTIVE: Arterial infusion chemotherapy for unresectable pancreatic cancer may be an attractive strategy. We are currently developing a new interventional radiology technique to unify the pancreatic blood supply for chemoinfusion. In this article, we evaluated the feasibility, therapeutic potential, and limitations of this technique. MATERIALS AND METHODS: Twenty-eight patients with advanced pancreatic cancer were retrospectively reviewed. After the superior mesenteric artery was embolized, the drug distribution to the tumor was evaluated by a unified CT angiography system. RESULTS: In all patients, tumors had dual arterial blood supplies from both the celiac and superior mesenteric arteries. Unification of pancreatic blood supply was successful in 14 patients (50%). The whole pancreatic tumor was exclusively supplied by the celiac artery in these patients. In the remaining 14 patients, a part of the tumor was still supplied by the superior mesenteric artery even after embolization. In most failed cases, the tumor had invaded to the root of the small-bowel mesentery or transverse mesocolon. By contrast, unification of the pancreatic blood supply could be achieved in cases invading to the stomach and duodenum. The whole liver was also successfully supplied by arterial infusion from the celiac artery after embolization. CONCLUSION: Our novel technique for arterial infusion chemotherapy is simple, feasible, and well tolerated. Furthermore, arterial chemoinfusion may be a breakthrough in selected patients with unresectable pancreatic cancer. PMID- 19304678 TI - Endoleaks after endovascular abdominal aortic aneurysm repair: management strategies according to CT findings. AB - OBJECTIVE: With increasing use of endovascular techniques for repair of abdominal aortic aneurysms, the prevalence of leakage into excluded aneurysm sacs (endoleaks) as a complication has risen. We will describe and illustrate the imaging findings for endoleaks involving abdominal aortic aneurysms. We will also discuss which types of endoleaks require urgent catheter-based evaluation. CONCLUSION: Radiologists should be familiar with the classification scheme for endoleaks and understand which types of endoleaks require urgent catheter-based evaluation. PMID- 19304679 TI - Probably benign breast masses diagnosed by sonography: is there a difference in the cancer rate according to palpability? AB - OBJECTIVE: The study was designed to assess retrospectively whether there is a difference in the cancer rates between palpable and nonpalpable probably benign breast nodules detected by sonography. We further investigated the clinicopathologic results of sonographically false-negative cases. MATERIALS AND METHODS: This study included 352 women who had undergone sonographically guided core biopsies for 374 BI-RADS category 3 masses between March 2004 and February 2005. No masses were diagnosed with definite malignant findings on mammography. The cancer rates of nonpalpable and palpable masses were compared using Fisher's exact test. The clinicopathologic results of sonographically false-negative cases were investigated. RESULTS: Among the 374 masses, 86 masses (23%) that were lost to follow-up were excluded. Of the 288 masses with follow-up or excision, the cancer rate was 2.4% (7/288). The cancer rate of the nonpalpable masses was 2.1% (4/194) (95% CI, 0.6-5.2%), and the cancer rate of the palpable masses was 3.2% (3/94) (95% CI, 0.6-9.0%), with no statistically significant difference (p = 0.6864). Of the seven cancers, five were diagnosed by a sonographically guided core biopsy and two were diagnosed by surgical excision after a benign biopsy. The seven sonographically false-negative masses in seven patients were identified as three invasive ductal carcinomas, two ductal carcinomas in situ, one mucinous carcinoma, and one papillary carcinoma on the basis of pathology results. CONCLUSION: There is no statistically significant difference between the cancer rates of palpable and nonpalpable BI-RADS category 3 masses seen on sonography. PMID- 19304680 TI - Administration of gadolinium-based contrast agents in MR angiography. PMID- 19304681 TI - Gadolinium and nephrogenic systemic fibrosis. PMID- 19304682 TI - Primary and postprimary or reactivation tuberculosis: time to revise confusing terminology? PMID- 19304683 TI - Communicating imaging results to patients: OnSite results. PMID- 19304684 TI - Medical physics--meeting the needs of radiologists: the role of the American Journal of Roentgenology. PMID- 19304685 TI - Teaching physics to radiology residents. AB - OBJECTIVE: The complexity of diagnostic imaging has expanded dramatically over the past two decades. Over the same period, the time and effort devoted to teaching physics (the science and technology of the discipline) have diminished. This paradox compromises the ability of future radiologists to master imaging technologies so that they are used in an efficient, safe, and cost-effective manner. This article addresses these issues. CONCLUSION: Efforts involving many professional organizations are under way to resolve the paradox of the expanding complexity of medical imaging contrasted with the declining emphasis on physics in radiology residency programs. These efforts should help to reestablish physics education as a core value in radiology residency programs. PMID- 19304686 TI - Teaching medical students about imaging techniques. AB - OBJECTIVE: This article reviews major themes about imaging techniques with which all educators should be familiar. CONCLUSION: A high-quality introduction to radiologic techniques should provide a basic framework to make good use of radiology in caring for patients. Tomorrow's physicians will need to make effective and efficient use of new imaging techniques. The responsibility for preparing them to do so rests squarely on the shoulders of today's radiology educators. PMID- 19304687 TI - Misoperation of CT automatic tube current modulation systems with inappropriate patient centering: phantom studies. AB - OBJECTIVE: Inappropriate patient centering on the gantry changes the size of the localizer radiographs used for CT examinations, influencing the operation of CT automatic tube current modulation because tube current is controlled with information from localizer radiographs. The purpose of this study was to examine the influence of inappropriate patient centering on the gantry isocenter on automatic tube current modulation. MATERIALS AND METHODS: An elliptical phantom was scanned with four automatic tube current modulation techniques after acquisition of localizer radiographs in the horizontal and vertical directions with the phantom center shifted from the gantry isocenter in the vertical direction. After scanning, the magnification rate of the frontal localizer radiographs, tube current-time product, and image noise were examined. RESULTS: On phantom studies, the magnification rate of localizer radiographs showed a linear relation to the vertical deviation of the phantom from the gantry isocenter. From 50 mm above to 50 mm below the gantry isocenter, tube current time products ranged from 75% to 141% compared with those at the gantry isocenter. In addition, increases and decreases in the amount of image noise related to changes in tube current-time product were confirmed. CONCLUSION: Inappropriate patient centering causes misoperation of automatic tube current modulation systems, in which tube current is controlled with information from localizer radiographs, and thus causes increases in tube current or image noise. PMID- 19304688 TI - Radiation dose in a "triple rule-out" coronary CT angiography protocol of emergency department patients using 64-MDCT: the impact of ECG-based tube current modulation on age, sex, and body mass index. AB - OBJECTIVE: "Triple rule-out" coronary CT angiography (CTA) using 64-MDCT technology is a new approach for evaluating emergency department patients presenting with symptoms suggestive of acute coronary syndrome (ACS). Our objective was to evaluate the reduction in effective radiation dose through the use of tube current modulation in patients who underwent a triple rule-out coronary CTA evaluation and to document how effective radiation dose was impacted by patient age, sex, and body mass index (BMI). MATERIALS AND METHODS: A retrospective analysis of triple rule-out coronary CTA examinations performed on a 64-MDCT scanner was ordered on a prospective cohort of 267 consecutive low- to moderate-risk emergency department patients with suspected ACS from a single university hospital between October 2006 and March 2008. Tube current modulation was generally used in patients with heart rates below 65 beats per minute during the second half of the study period as a way to reduce radiation exposure. We calculated effective radiation exposure using actual patient coronary CTA scanning parameters by age, sex, and BMI. RESULTS: Among the 172 patients evaluated without tube current modulation, effective dose averaged (+/- SD) 18.0 +/- 5.6 mSv (range, 9.9-31.3 mSv). Of the 95 patients who underwent CTA examination with tube current modulation, effective dose was significantly lower at 8.75 +/- 2.64 mSv (range, 5.4-16.6 mSv; p < 0.0001) and image quality was better (p < 0.0001) as compared with examinations without tube current modulation. There were no significant radiation differences by patient age, but tube current modulation decreased radiation exposure by at least half. Among the studies in which tube current modulation was not used, women received less radiation than men (17.0 vs 19.5 mSv, respectively; p < 0.001). For the studies with tube current modulation, there were no radiation differences by sex. Obese patients received significantly more radiation than overweight and normal-weight patients in the non-tube current modulation groups (20.9 mSv vs 15.0 and 14.9 mSv, respectively; p < 0.0001) and in the tube current modulation groups (10.3 mSv vs 7.6 and 7.1 mSv, p < 0.0001). CONCLUSION: The overall effective radiation dose for triple rule-out coronary CTA was reduced by more than 50% with ECG-based tube current modulation without loss of image quality. Tube current modulation should be used for triple rule-out coronary CTA examinations whenever possible. PMID- 19304689 TI - MRI of the spine: image quality and normal-neoplastic bone marrow contrast at 3 T versus 1.5 T. AB - OBJECTIVE: The objectives of our study were to compare image quality of the spine and visualization of spine abnormalities at 3 T and 1.5 T as well as to evaluate differences in quantitative assessment of normal and neoplastic vertebral bone marrow. MATERIALS AND METHODS: One hundred nine MR examinations of the spine were performed at 1.5 T and 3 T in the same patients within a time interval of less than 3 months. Visualization of anatomic and pathologic structures was analyzed by two radiologists. Normal and pathologic bone marrow was assessed on T1 weighted fast spin-echo (FSE) sequences. The signal intensity contrast of neoplastic bone marrow versus normal vertebral bone marrow was measured at 1.5 T versus 3 T. Sensitivity, specificity, and accuracy with 95% CIs were computed to assess the performance of muscle and disk as standards to differentiate between neoplastic and normal bone marrow on T1-weighted sequences at 1.5 T and 3 T. RESULTS: For all anatomic structures evaluated, image quality was rated significantly higher at 3 T than at 1.5 T, with 71.6% of the studies overall being superior at 3 T. The contrast between normal and pathologic bone marrow was significantly larger at 3 T (mean +/- SD, 0.33 +/- 0.13) than at 1.5 T (0.27 +/- 0.11). The highest accuracy was found using muscle signal at 3 T to differentiate between normal and pathologic bone marrow. CONCLUSION: The use of 3-T MRI improves visualization of anatomic structures in the spine over 1.5-T MRI. As an internal standard on T1-weighted FSE images, skeletal muscle can be used to differentiate between infiltrative and normal bone marrow with higher accuracy at 3 T than at 1.5 T. PMID- 19304690 TI - Assessment of patient and occupational dose in established and new applications of MDCT fluoroscopy. AB - OBJECTIVE: This study aimed to assess patient dose and occupational dose in established and new applications of MDCT fluoroscopy. MATERIALS AND METHODS: Electronic personal dosimeters were used to measure occupational dose equivalent. Effective patient dose was derived from the recorded dose-length product. Acquisition parameters that were observed during CT fluoroscopy (CTF) provided the basis for the estimation of an entrance skin dose profile. Two hundred ten CT guided interventional procedures were included in the study. RESULTS: The median effective patient dose was 10 mSv (range, 0.1-235 mSv; 107 procedures). The median peak entrance skin dose was 0.4 Sv (0.1-2.1 Sv; 27 procedures). From 547 measurements of occupational dose equivalent, a median occupational effective dose of 3 muSv per procedure was derived for the interventional radiologists and 0.4 muSv per procedure for the assisting radiologists and radiology technologists. The estimated maximum occupational effective dose reached 0.4 mSv. CONCLUSION: The study revealed high effective patient doses, up to 235 mSv, mainly for relatively new applications such as CTF-guided radiofrequency ablations using MDCT, vertebroplasty, and percutaneous ethanol injections of tumors. Entrance doses were occasionally in the range of the warning level for deterministic skin effects but were always below the threshold for serious deterministic effects. The complexity of the procedure, expected benefits of the treatment, and general health state of the patient contribute to the justification of observed high effective patient doses. PMID- 19304691 TI - Cumulative radiation exposure and cancer risk estimates in emergency department patients undergoing repeat or multiple CT. AB - OBJECTIVE: The purpose of our study was to define a conservative estimate of the number of patients undergoing repeat or multiple emergency department CT studies and to quantify their cumulative CT radiation doses and lifetime attributable risk of developing cancer. MATERIALS AND METHODS: We identified all patients at a tertiary care adult academic medical center with at least three emergency department visits within a 1-year period that included CT of the neck, chest, abdomen, or pelvis. For this cohort, we identified all diagnostic CT studies over the previous 7.7 years. We calculated cumulative radiation doses by summing typical effective doses of the anatomic regions scanned, and we calculated lifetime attributable risk using the population-averaged dose-to-risk conversion factor of one cancer per 1,000 patients receiving a 10-mSv dose, in accordance with the seventh Biologic Effects of Ionizing Radiation (BEIR VII) report. RESULTS: One hundred thirty emergency department patients met the inclusion criteria. Over the 7.7-year period, median, mean, and maximum values for the study count were 10, 13, and 70 with cumulative CT doses of 91, 122, and 579 mSv and lifetime attributable risk of one in 110, one in 82, and one in 17, respectively. Emergency department studies comprised 55% of those captured. Repeat imaging of the same study type represented at least half of the imaging for 72% of the cohort and all of the imaging for 12%. CONCLUSION: A small proportion (1.9%) of emergency department patients undergoing CT of the neck, chest, abdomen, or pelvis have high cumulative rates of multiple or repeat imaging. Collectively, this patient subgroup may have a heightened risk of developing cancer from cumulative CT radiation exposure. PMID- 19304692 TI - MDCT for suspected appendicitis: effect of reconstruction section thickness on diagnostic accuracy, rate of appendiceal visualization, and reader confidence using axial images. AB - OBJECTIVE: The purpose of this study was to evaluate interpretative performance with different MDCT reconstruction parameters in adult patients with suspected appendicitis. MATERIALS AND METHODS: MDCT scans of 212 adult patients obtained in an emergency department with a 64-MDCT scanner were prospectively collected. Acquisition technique included 24 x 1.2 mm detector configuration and IV contrast administration with or without oral contrast administration. Data sets were reconstructed with three techniques: 5 x 5 mm, 3 x 3 mm, and 2 x 1 mm (section thickness x interval). Each of the 212 sets of images (grouped by reconstruction technique) was reviewed retrospectively using axial sections by two independent readers blinded to diagnosis. Medical record review was conducted to identify patients with appendicitis. Visualization of the appendix, confidence in visualization, confidence for presence or absence of specific CT findings, diagnostic accuracy, and diagnostic confidence were compared across reconstruction techniques. Data were analyzed with simple and ordinal logistic regression with adjustment for multiple observations derived from each patient and for reader differences. RESULTS: Progressively thinner reconstruction section thickness was associated with a significant increase in the rate of visualization of the appendix (p < 0.001 for 5 x 5 vs 3 x 3; p = 0.03 for 3 x 3 vs 2 x 1), visualization confidence (p < 0.001 for 5 x 5 vs 3 x 3 and 3 x 3 vs 2 x 1), and confidence for presence or absence of findings. Seventeen subjects (8%) had appendicitis. Correctness of diagnosis was not significantly associated with reconstruction method. However, for correctly diagnosed cases interpreted as normal, impression confidence increased with progressively thinner section thickness (p < 0.001 for 5 x 5 vs 3 x 3 and 3 x 3 vs 2 x 1). CONCLUSION: In this investigation of contrast-enhanced MDCT of the appendix, visual ization of the appendix and confidence in interpretation of axial images progressively improved with use of thinner reconstruction sections. PMID- 19304693 TI - Helical CT evaluation of the preoperative staging of gastric cancer in the remnant stomach. AB - OBJECTIVE: The objective of our study was to evaluate the diagnostic performance of helical CT for evaluating the preoperative staging of gastric cancer in the remnant stomach. MATERIALS AND METHODS: Preoperative helical CT images of 67 patients with gastric cancer in the remnant stomach were independently analyzed regarding staging of the tumor by two radiologists who were blinded to histopathologic and surgical results. The differences in their assessments were resolved by consensus including the opinion of a third radiologist. The radiologists were asked to determine the depth of tumor invasion of the gastric wall (T stage), classifying it as 5%. The area under the receiver operating characteristic curve was 99.2% (95% CI, 98-100%). The optimal hepatorenal sonographic index cutoff point for the prediction of steatosis > 5% was 1.49, with sensitivity of 100% and specificity of 91%. The optimal hepatorenal sonographic index cutoff point for the prediction of steatosis >/= 25% was 1.86, with sensitivity of 90% and specificity of 90%. The optimal hepatorenal sonographic index cutoff point for the prediction of steatosis >/= 60% was 2.23, with sensitivity of 90% and specificity of 93%. CONCLUSION: The hepatorenal sonographic index is a sensitive noninvasive method for steatosis quantification. It can diagnose small amounts of liver fat that would be missed by conventional sonography. It is reproducible and operator independent and can serve as an efficient tool to follow patients with steatosis and evaluate the efficacy of new treatment techniques. PMID- 19304695 TI - Respiratory-triggered versus breath-hold diffusion-weighted MRI of liver lesions: comparison of image quality and apparent diffusion coefficient values. AB - OBJECTIVE: The purpose of our study was to compare respiratory-triggered and breath-hold diffusion-weighted liver MRI and to assess the agreement in the apparent diffusion co-efficient (ADC) values between the two sequences. MATERIALS AND METHODS: Forty-eight patients (27 men, 21 women; mean age, 45.2 years) with focal liver lesions underwent respiratory-triggered and breath-hold diffusion weighted MRI (DWI) in addition to routine MRI. Both sequences had identical imaging parameters except for signal averages, which were 6 in respiratory triggered and 2 in breath-hold sequences. A total of 92 lesions (maximum of three lesions per patient; 37 benign, 55 malignant) were evaluated. Lesions were confirmed by typical imaging appearance, histopathology, or follow-up. Signal-to noise ratio (SNR) of the liver, contrast-to-noise ratio (CNR), and relative contrast ratio of the lesions were measured in each DWI sequence and were statistically compared using the Mann-Whitney U test. The ADC values of normal liver and each category of liver lesions in the two sequences were compared for agreement using Pearson's coefficient and reliability analysis scale. RESULTS: The SNR of the normal liver was significantly better on respiratory-triggered DWI than on breath-hold DWI. The mean CNR of metastases, hepatocellular carcinomas, and abscesses was significantly better in the respiratory-triggered DWI than in the breath-hold DWI sequences. The ADC values of liver and focal lesions measured by the two techniques showed good agreement. The SDs of the ADC values of normal liver were similar in the two sequences. CONCLUSION: Respiratory-triggered DWI should be preferred over breath-hold DWI for the evaluation of focal liver lesions because it provides better image quality and SNR without any compromise in the calculated ADC values. PMID- 19304696 TI - The bloody pancreas: MDCT and MRI features of hypervascular and hemorrhagic pancreatic conditions. AB - OBJECTIVE: The purpose of this study was to illustrate the varied MDCT and MRI appearances of hypervascular and hemorrhagic pancreatic conditions and their mimics. CONCLUSION: Pancreatic hypervascular conditions are easily detected at multiphasic contrast-enhanced MDCT and MRI. Hemorrhagic pancreatic abnormalities are best depicted with unenhanced CT and, especially, fat-suppressed T1-weighted MRI. Familiarity with the spectrum of possible underlying causes and the imaging features and conditions that can act as mimics assists radiologists in making an accurate presumptive diagnosis. PMID- 19304697 TI - Scirrhous metastases to the gastrointestinal tract at CT: the malignant target sign. AB - OBJECTIVE: Our objective was to study the presence of a characteristic appearance of metastatic disease to the gastrointestinal tract on contrast-enhanced CT in patients with known malignancies and to investigate its clinical implications. CONCLUSION: Twenty-five patients with scirrhous metastases had a malignant CT target sign. Careful observation and correlation with clinical history are required to differentiate this unique sign from a benign target sign. PMID- 19304698 TI - Spatial resolution and radiation dose of a 64-MDCT scanner compared with published CT urography protocols. AB - OBJECTIVE: The objective of our study was to compare the spatial resolution and effective dose from 64-MDCT with several published CT urography protocols. MATERIALS AND METHODS: A phantom containing 1-, 2-, or 4-mm cylindric channels to simulate ureters with 0.25- to 3-mm plugs to simulate ureteral filling defects or ureteral diverticula was imaged using eight helical CT urography protocols. Computed radiography (CR) was also performed. Coronal maximum-intensity projection images were created and, with the CR image, were evaluated independently by two genitourinary radiologists. Spatial resolution was evaluated by scoring each abnormality as present, visible; or as absent, not visible. Effective dose estimates for 11 CT urography protocols, including the radiographs obtained in the CT urography protocol, were calculated using published Monte Carlo organ dose coefficients. RESULTS: All ureteral abnormalities detected on CR were detected on the highest-spatial-resolution reconstruction using the evaluated 64-MDCT system. The smallest filling defect identified by both was 0.25 mm. Three 0.25-mm filling defects were not detected using the evaluated 16-MDCT system. The 4-MDCT system protocols showed the poorest performance. The range of effective doses for the evaluated CT urography protocols was 20.1-66.3 mSv. The number of phases, anatomic coverage per phase, and scanning parameters all contributed to this variation in dose. CONCLUSION: The evaluated 64-MDCT system showed detection accuracy identical to that of CR. Limiting anatomic coverage for specific phases and combining phases can reduce dose for multiphase protocols by up to a factor of 2 relative to early (circa 2000) 4-MDCT. PMID- 19304699 TI - Value of dynamic contrast-enhanced MRI and correlation with tumor angiogenesis in bladder cancer. AB - OBJECTIVE: The aim of this study was to investigate dynamic contrast-enhanced MRI (DCE-MRI) for the noninvasive measurement of bladder cancer angiogenesis by correlation with microvessel density, histologic grade, and tumor staging, and to predict the outcome of local recurrence. MATERIALS AND METHODS: Twenty-four patients with bladder cancer were examined using DCE-MRI. Hemodynamic parameters obtained by DCE-MRI included peak time enhancement in the first minute (E(max/1)) after contrast administration, second minute (E(max/2)), third minute (E(max/3)), fourth minute (E(max/4)), and fifth minute (E(max/5)), and the steepest slope. Microvessel density was identified by immunostaining of endothelial cells using FVIII-related antigen. The Mann-Whitney U test, multivariate discriminant analysis, Spearman's correlation coefficient, and analysis of variance were used for statistical analysis. RESULTS: Correlation was seen between DCE-MRI parameters (E(max/1) and steepest slope) and microvessel density (p < 0.05). E(max/1) and steepest slope were found to have a statistically significant correlation with histologic grade (p < 0.05 and p < 0.01, respectively). A significant difference was seen between groups of patients with and without local recurrence with regard to two of the DCE-MRI parameters (p < 0.05 for E(max/1) and E(max/2)). CONCLUSION: The contrast enhancement patterns on DCE-MRI are influenced by tumor angiogenesis, as reflected by elevated microvessel density expression. Therefore, they are valuable indicators for assessing tumor angiogenic activity and tumor neovascularization in bladder cancers. PMID- 19304700 TI - PET/CT for the characterization of adrenal masses in patients with cancer: qualitative versus quantitative accuracy in 150 consecutive patients. AB - OBJECTIVE: The objective of our study was to evaluate a large cohort of patients with PET/CT to determine whether qualitative (visual) assessment, quantitative standardized uptake value (SUV), or standardized uptake ratio (SUR) techniques should be used when attempting to characterize adrenal masses in patients with cancer. MATERIALS AND METHODS: The study group was composed of 150 consecutive patients (78 men, 72 women; mean age, 60 years; range, 24-88 years) with documented adrenal lesions. All patients were known to have an underlying primary malignancy and were referred for PET/CT to evaluate the underlying primary and metastatic tumor burden. Definitive lesion characterization was determined by evaluating all histologic adrenal specimens and all relevant prior and follow-up CT scans, including unenhanced, contrast-enhanced, and delayed contrast-enhanced washout studies. RESULTS: Of the 139 benign lesions, 109 were considered benign by CT densitometry measurements and 135 by qualitative PET data. Qualitative PET characterized 28 of 30 benign lesions that were considered indeterminate by unenhanced CT. All 26 malignant lesions were characterized by PET: All showed qualitative and quantitative signal intensity greater than the liver. By combining unenhanced and qualitative CT data with the retrospective PET data, the analysis yielded a sensitivity of 100% for the detection of malignancy, a specificity of 99%, a positive predictive value (PPV) of 93%, a negative predictive value (NPV) of 100%, and an accuracy of 99% (Table 1). Conversely, for the detection of benignity, the sensitivity, specificity, PPV, NPV, and accuracy were 99%, 100%, 100%, 93%, and 99%, respectively. CONCLUSION: PET/CT is a highly accurate method for differentiating benign from malignant adrenal masses particularly when using qualitative, rather than quantitative, PET data. The routine use of quantitative mean or maximal SUV or SUR data may be unnecessary. Occasional benign lesions do show mild to moderate increased FDG uptake compared with that of the liver and may mimic some malignant lesions. Without evidence that these lesions are benign by unenhanced CT densitometry or adrenal mass stability or growth from previous CT scans, we recommend that these lesions be characterized using contrast-enhanced washout tests and that if those tests are inconclusive, using percutaneous biopsy if early lesion characterization is mandatory. PMID- 19304701 TI - CT features of adnexal involvement in patients with diverticulitis. AB - OBJECTIVE: The purpose of this study was to evaluate the CT findings in patients with colosalpingeal fistula complicating diverticulitis. MATERIALS AND METHODS: Using a medical records database and radiology information system, we identified the cases of 14 women who underwent colonic resection for diverticulitis and simultaneous salpingo-oophorectomy with or without hysterectomy over a 10-year period. The patients were separated into two groups: group 1 included women with a pathologically proven colosalpingeal fistula, and group 2 included those who had contiguous periadnexal inflammation without a fistula. Preoperative CT scans of these patients were retrospectively evaluated in consensus by two radiologists blinded to the pathologic findings. The presence of adnexal gas and fluid collections was recorded. The findings were compared with those in the operative and pathology reports. RESULTS: Among 14 patients who had sigmoid (n = 13) or cecal (n = 1) diverticulitis, eight patients were in group 1 and six were in group 2. On CT scans, an adnexal collection of gas, either alone or in combination with fluid, was found in seven patients (88%) in group 1 and in no patients in group 2, resulting in 88% sensitivity and 100% specificity in the detection of colosalpingeal fistula. An adnexal collection of fluid alone was found in one patient (13%) in group 1 and one patient (17%) in group 2, resulting in sensitivity and specificity of 13% and 83%. CONCLUSION: In patients with diverticulitis, a collection of gas, either alone or in combination with fluid, within the adnexum is a sensitive and specific predictor of the presence of a fistula between the colon and an adnexum. PMID- 19304702 TI - Ligaments of the posterior and lateral talar processes: MRI and MR arthrography of the ankle and posterior subtalar joint with anatomic and histologic correlation. AB - OBJECTIVE: MRI and combined ankle and posterior subtalar MR arthrography in cadavers were used to evaluate the ligaments of the posterior and lateral talar processes. Subsequent anatomic and histologic correlation was performed. MATERIALS AND METHODS: Ten cadaveric ankles were used. Routine radiography and MRI were initially performed. Ankle and posterior subtalar MR arthrography, followed by anatomic and histologic analysis, was then performed to allow better assessment of the ligaments of the lateral and posterior talar process. RESULTS: In all subjects, MR arthrography provided superior delineation of the articular and periarticular structures, as well as the ligaments. The lateral talocalcaneal and medial talocalcaneal ligaments were best seen in the axial and coronal planes, respectively. The axial plane was best for visualizing the fibulotalocalcaneal ligament, and the sagittal plane was best for evaluating the posterior talocalcaneal ligament. The anterior and posterior talofibular ligaments and the posterior tibiotalar ligament (superficial and deep portions) were best seen in the axial plane. Histologic analysis was correlated to anatomic sectioning and showed the attachment sites of these ligaments. CONCLUSION: Combined ankle and posterior subtalar MR arthrography enhances visualization of the ligaments attaching to the posterior and lateral talar processes, including the posterior, lateral, and medial talocalcaneal and fibulotalocalcaneal ligaments. PMID- 19304703 TI - Intervertebral disk degeneration related to reduced vertebral marrow perfusion at dynamic contrast-enhanced MRI. AB - OBJECTIVE: The purpose of this study was to use dynamic contrast-enhanced MRI to ascertain the relation between intervertebral disk degeneration and lumbar vertebral marrow blood perfusion. SUBJECTS AND METHODS: We recruited 25 patients (50 vertebral bodies) who underwent dynamic contrast-enhanced MRI of the lumbar spine. The peak signal enhancement of each vertebral body was calculated from the time signal after curve fitting of a pharmacokinetic model. We controlled for other variables that might have affected blood perfusion by assessing two vertebral bodies in each patient. The 25 patients were divided into three groups. In group 1, one of the vertebral bodies (L1 or L3) evaluated was between two adjacent normal disks and the other was between two adjacent degenerated disks. In group 2, each of the two vertebral bodies evaluated was between two normal disks. In group 3 each of the two vertebral bodies evaluated was between two degenerated disks. RESULTS: Without normalization by minimization of other variables, there were no statistically significant differences in original peak enhancement values among groups 1, 2, and 3 (p = 0.179). After normalization, the peak enhancement in group 1 (0.846 +/- 0.060) was significantly lower than that in group 2 (0.988 +/- 0.047) (p = 0.003) or group 3 (0.973 +/- 0.081) (p = 0.008). CONCLUSION: After normalization, lumbar vertebral marrow perfusion correlated well with intervertebral disk degeneration evaluated with dynamic contrast-enhanced MRI. Blood perfusion was 14% less in the vertebral body marrow between two degenerated disks than in vertebral marrow between two normal disks. PMID- 19304704 TI - Whole-Body MRI versus PET in assessment of multiple myeloma disease activity. AB - OBJECTIVE: The purpose of this study was to compare FDG PET; whole-body MRI; and the reference standard, bone marrow aspiration and biopsy, to determine the best imaging technique for assessment of disease activity in multiple myeloma. SUBJECTS AND METHODS: Twenty-four patients (13 women, 11 men; mean age, 67.1 years; range, 44-83 years) with bone marrow biopsy-proven multiple myeloma were included in the study. All patients underwent PET/CT and whole-body MRI within 10 days of each other. Eight patients underwent more than one follow-up PET/CT and whole-body MRI examination, for a total of 34 pairs of images. Activity was defined as lesions with a maximum standardized uptake value greater than 2.5 at PET and as evidence of marrow packing at whole-body MRI. PET and whole-body MRI results were correlated with findings at bone marrow aspiration biopsy. RESULTS: PET had a sensitivity of 59%, specificity of 75%, positive predictive value of 81%, and negative predictive value of 50% (p = 0.08). Whole-body MRI had a sensitivity of 68%, specificity of 83%, positive predictive value of 88%, and negative predictive value of 59% (p = 0.01). In 62% of cases, PET and whole-body MRI findings were concordant. When PET and whole-body MRI findings were concordant and positive, no false-positive results were found, yielding a specificity and a positive predictive value of 100% (p = 0.04). CONCLUSION: Whole body MRI performed better than PET in the assessment of disease activity, having a higher sensitivity and specificity. The positive predictive value of whole-body MRI in the assessment of active disease was high at 88%. When used in combination and with concordant findings, PET and whole-body MRI were found to have a specificity and positive predictive value of 100%, which may be of value to clinicians assessing the effectiveness of aggressive and expensive treatment regimens. PMID- 19304705 TI - Fractures of the ankylosed spine: MDCT and MRI with emphasis on individual anatomic spinal structures. AB - OBJECTIVE: The purpose of this article is to illustrate the spectrum of MDCT and MRI appearances of spinal fractures in ankylosing spondylitis and diffuse idiopathic skeletal hyperostosis. CONCLUSION: Spinal fractures associated with ankylosing spondylitis and diffuse idiopathic skeletal hyperostosis usually involve the three columns of the spine, and injury to the posterior osteoligamentous component is the hallmark of these fractures. Osseous and ligamentous injuries can be accurately visualized and analyzed with MDCT with multiplanar reformation and with MRI. PMID- 19304706 TI - Initial experience with imaging-guided intramuscular botulinum toxin injection in patients with idiopathic cervical dystonia. AB - OBJECTIVE: The objective of our study was to present our initial experiences of imaging-guided intramuscular botulinum toxin (BTX) injection in patients with idiopathic cervical dystonia. CONCLUSION: Imaging-guided BTX injection is a useful treatment technique in patients with idiopathic cervical dystonia when target muscles are located deeply or an injection must be delivered to a focal muscle area. PMID- 19304707 TI - CNS complications of hematopoietic stem cell transplantation. AB - OBJECTIVE: With the worldwide increase in the use of hematopoietic stem cell transplantation (HSCT), a high level of diligence is required for radiologists to understand HSCT-related complications in the CNS. This article describes the clinical background of HSCT and complications that occur in a time-dependent manner through the course of HSCT and addresses pivotal issues in diagnostic imaging. CONCLUSION: Acknowledging the realm of imaging manifestations and the underlying mechanism of HSCT will enhance diagnostic accuracy and optimize treatment decisions. PMID- 19304708 TI - Whole-body MRI and FDG PET fused images for evaluation of patients with cancer. AB - OBJECTIVE: The purpose of this article is to illustrate the feasibility of fused whole-body MRI and PET for the evaluation of patients with cancer. CONCLUSION: MRI has proven to be superior to CT for studying several regions of the body, and PET FDG has become an established diagnostic tool in oncology. Because FDG accumulates avidly in tumor tissue, fusing FDG PET to whole-body MRI allows good anatomy-based evaluation of disease. PMID- 19304709 TI - Molecular imaging and quantitative measurement of epidermal growth factor receptor expression in live cancer cells using immunolabeled gold nanoparticles. AB - OBJECTIVE: The goal of this study was to assess whether immunolabeled nanoparticle biomarkers are comparable to fluorescent marker imaging in measuring epidermal growth factor receptor (EGFR) expression. MATERIALS AND METHODS: EGFR expression was quantified using both imaging methods in four cell lines: A431 human epidermoid carcinoma cells, which are known to have high EGFR expression; two cell lines with lower EGFR expression (270-GBM human glioblastoma xenograft cells and H2224 human glioblastoma xenograft cells); and MDA-MB-453 breast carcinoma cells, which do not express EGFR. To enhance contrast of the nanoparticle biomarkers, a darkfield microspectroscopy system was used that includes a custom epi-illumination light train. RESULTS: Nanoparticle-bound cells were clearly distinguished from control cells not bound to nanoparticles in that they showed a significant increase in detected intensity under darkfield illumination due to nanoparticle scattering. The average nanoparticle-scattering intensity for A431 cells was 41.5 counts per cell compared with 24.7 for 270-GBM cells, 8.77 for H2224 cells, and 0.44 for MDA-MB-453 cells. The average fluorescence intensity for A431 cells was 35.3 counts per cell compared with 28.7 for 270-GBM cells, 5.91 for H2224 cells, and 2.07 for MDA-MB-453 cells. A plot of fluorescence intensity versus nanoparticle-scattering intensity for all four cell lines showed that the data agree with a linear relationship given by the following equation: NP = 1.0691 x FL - 0.3873, where NP is the nanoparticle scattering intensity and FL is the fluorescence intensity. The covariance of the data with the trend line was R(2) = 0.9409. The average peak wavelength of nanoparticle scattering was 570.93 nm for A431 cells, 565.26 nm for 270-GBM cells, and 562.70 nm for H2224 cells (with no clear peaks observed for MDA-MB-453 cells). This spectral trend shows that nanoparticle scattering may reveal additional information about their nanoenvironment via refractive index sensitivity. CONCLUSION: Immunolabeled nanoparticles can quantify receptor expression with performance comparable to fluorescence markers and show promise to better characterize receptor expression via their refractive index sensitivity. PMID- 19304710 TI - Spectrum of focal benign musculoskeletal 18F-FDG uptake at PET/CT of the shoulder and pelvis. AB - OBJECTIVE: The purpose of this article is to illustrate the spectrum of common benign intraarticular and extraarticular disorders associated with focal (18)F FDG uptake in the shoulder and pelvic areas in oncology patients referred for PET/CT. CONCLUSION: A wide spectrum of benign musculoskeletal disorders associated with focal FDG uptake may be detected in cancer patients. This incidental uptake usually does not seem to be a clinically significant finding, but it can affect quality of life. PMID- 19304711 TI - Automated algorithm for quantifying the extent of cystic change on volumetric chest CT: initial results in Lymphangioleiomyomatosis. AB - OBJECTIVE: The purpose of our study was to develop a new method for quantifying the severity of cystic lung disease using chest CT and to evaluate this method in patients with lymphangioleiomyomatosis (LAM). SUBJECTS AND METHODS: Eighteen patients with LAM (all women; mean age, 43.6 years) underwent chest CT and pulmonary function testing including diffusing capacity for carbon monoxide (DLCO). All patients were at their clinical baseline on the day of imaging. Standard quantitative CT metrics including the percentage of the lung volume < 910 HU and the 15th percentile of Hounsfield units were computed from the histogram of lung voxels. A new histogram analysis method was developed to compute the cyst volume and the volume of the remaining lung by segmenting the entire lung attenuation histogram into two underlying distributions, one from the cysts and the other from the remaining lung tissue. RESULTS: The mean +/- SD for quantitative lung metrics was 21% +/- 16% for percentage < -910 HU, -915 +/- 47 HU for 15th percentile of Hounsfield units, and 19% +/- 13% for cyst volume. The correlation between pulmonary function tests and CT metrics was strongest for the percentage of cyst volume for all pulmonary function testing indexes, with correlations between forced expiratory volume in 1 second (FEV(1)) percentage predicted and the CT metrics of r = -0.52, r = 0.50, and r = -0.86 for the percentage of lung < -910 HU, the 15th percentile of Hounsfield units, and the percentage of cyst volume, respectively. CONCLUSION: A new method for quantifying cyst volume as a percentage of total lung volume using chest CT correlates with pulmonary function parameters in patients with LAM and may have utility in the assessment of disease severity and progression of cystic lung diseases. PMID- 19304712 TI - Image quality in a low radiation exposure protocol for retrospectively ECG-gated coronary CT angiography. AB - OBJECTIVE: The purpose of our study was to systematically compare the image quality of dual-source CT coronary angiography using 100 kV instead of 120 kV. SUBJECTS AND METHODS: One hundred patients with a body weight /= 50% stenosis or occlusion on MDCT angiography was calculated using DSA as the standard of reference. Wilcoxon's signed rank test and Mann-Whitney U test were used to compare differences in image quality between MDCT angiography and DSA and between MDCT angiography with the patient's arm stretched overhead or alongside the body, respectively. RESULTS: Interobserver agreement for detecting stenosis was excellent for both DSA (kappa = 0.86; 95% CI, 0.81-0.91) and MDCT angiography (kappa = 0.82; 95% CI, 0.77-0.87). Accuracy, sensitivity, specificity, PPV, and NPV of MDCT angiography for detecting >/= 50% stenosis or occlusion was 92.0% (95% CI, 86.8-95.3%), 90.2% (77.8-96.3%), 92.8% (85.9-96.6%), 85.2% (72.3-92.9%), and 95.4% (89.0-98.3%), respectively. No significant difference in image quality was seen between MDCT angiography and DSA (p = 0.3008) or between MDCT angiography with the patient's arm stretched overhead or alongside the body (p = 0.2912). CONCLUSION: MDCT angiography is a reproducible and reliable imaging technique for detection of >/= 50% stenosis or occlusion in dysfunctional hemodialysis fistulas. PMID- 19304718 TI - Fluoroscopic and sonographic guidance to place peritoneal catheters: how we do it. AB - OBJECTIVE: The purpose of this article is to detail a percutaneous approach to placing peritoneal catheters using sonographic and fluoroscopic guidance. Fluoroscopic-assisted placement of these catheters has been previously described in the literature. We emphasize the use of additional sonographic guidance, including color Doppler sonography, to determine the safest puncture site and to guide the initial needle puncture to avoid bowel perforation and injury to the epigastric artery. CONCLUSION: Imaging-guided-that is, sonography plus fluoroscopy-percutaneous placement of peritoneal catheters is a safe, minimally invasive, and effective alternative to blind or open surgical placement. The use of sonographic guidance together with fluoroscopic assistance makes placement of peritoneal catheters a safer and effective alternative to blind or open surgical placement. PMID- 19304719 TI - Percutaneous implantation of fiducial markers for imaging-guided radiation therapy. AB - OBJECTIVE: The use of imaging-guided radiation therapy (IGRT) to treat thoracic and abdominal tumors is increasing. In this article, we review the process of IGRT and describe techniques to implant fiducial markers in the optimal geometry. CONCLUSION: Implantation of fiducial markers can be challenging. A better understanding of the physics of IGRT can help optimize fiducial marker placement for precise tumor targeting. PMID- 19304720 TI - Mesenteric arterial variations detected at MDCT angiography of abdominal aorta. AB - OBJECTIVE: The purposes of this essay are to outline MDCT angiographic techniques for the evaluation of the mesenteric arterial vasculature and to review anatomic variants depicted on MDCT angiograms. CONCLUSION: MDCT angiography has distinct advantages over conventional angiography in imaging of the mesenteric arterial vasculature. PMID- 19304721 TI - Treatment of ureteropelvic junction obstruction using a detachable inflatable stent: initial experience. AB - OBJECTIVE: We describe a new method for treating ureteropelvic junction (UPJ) obstruction using a detachable inflatable stent positioned via a cystoscopic transvesicular approach. CONCLUSION: Eleven patients with UPJ obstruction were treated using a detachable inflatable stent, 64% of whom experienced complete pain relief. In 82% of patients, no obstruction was seen on renograms obtained after the procedure. The initial results achieved in the treatment of UPJ obstruction with a detachable inflatable balloon are promising, but further research in a larger patient population is required. PMID- 19304722 TI - Adenomyosis: sonohysterography with MRI correlation. AB - OBJECTIVE: The purpose of this study was to describe the sonohysterographic features of adenomyosis with MRI correlation. CONCLUSION: In this study, when the sonohysterographic findings suggested adenomyosis, MRI findings confirmed the diagnosis in 96% of cases. Myometrial cracks are, to our knowledge, a previously undescribed sonohysterographic sign of adenomyosis. PMID- 19304723 TI - A logistic regression model based on the national mammography database format to aid breast cancer diagnosis. AB - OBJECTIVE: The purpose of our study was to create a breast cancer risk estimation model based on the descriptors of the National Mammography Database using logistic regression that can aid in decision making for the early detection of breast cancer. MATERIALS AND METHODS: We created two logistic regression models based on the mammography features and demographic data for 62,219 consecutive mammography records from 48,744 studies in 18,269 [corrected] patients reported using the Breast Imaging Reporting and Data System (BI-RADS) lexicon and the National Mammography Database format between April 5, 1999 and February 9, 2004. State cancer registry outcomes matched with our data served as the reference standard. The probability of cancer was the outcome in both models. Model 2 was built using all variables in Model 1 plus radiologists' BI-RADS assessment categories. We used 10-fold cross-validation to train and test the model and to calculate the area under the receiver operating characteristic curves (A(z)) to measure the performance. Both models were compared with the radiologists' BI-RADS assessments. RESULTS: Radiologists achieved an A(z) value of 0.939 +/- 0.011. The A(z) was 0.927 +/- 0.015 for Model 1 and 0.963 +/- 0.009 for Model 2. At 90% specificity, the sensitivity of Model 2 (90%) was significantly better (p < 0.001) than that of radiologists (82%) and Model 1 (83%). At 85% sensitivity, the specificity of Model 2 (96%) was significantly better (p < 0.001) than that of radiologists (88%) and Model 1 (87%). CONCLUSION: Our logistic regression model can effectively discriminate between benign and malignant breast disease and can identify the most important features associated with breast cancer. PMID- 19304724 TI - Utility of targeted sonography for breast lesions that were suspicious on MRI. AB - OBJECTIVE: Data remain sparse to guide the use of targeted sonography for suspicious breast lesions found with MRI. The purpose of our study was to determine the usefulness of sonography in this setting. MATERIALS AND METHODS: Retrospective review of our breast MRI database was performed to identify all nonpalpable, mammographically occult MRI-detected suspicious lesions from January 1, 2003, to December 30, 2004, that underwent targeted sonography followed by imaging-guided biopsy. Frequency of sonography detection was determined, and MRI features (BI-RADS lesion type and size), histopathology outcomes, and positive biopsy rates of sonographically detected and occult findings were compared using chi-square analysis. RESULTS: Targeted sonography was performed for 167 of 201 MRI lesions. Outcomes were benign in 120 (72%) and malignant in 47 (28%) findings. Lesions assessed on sonography were 84 (50%) masses, 53 (32%) nonmasslike enhancements, and 30 (18%) foci, with mean size of 16 mm. Overall, 76 of 167 (46%) lesions were detected with sonography. Depiction was significantly more frequent for masses (58%) than for nonmasslike enhancement (30%) or foci (37%) (p = 0.003). Lesion size did not affect the frequency of detection. Sonographically depicted findings had a significantly higher positive biopsy rate (36%) than sonographically occult lesions (22%) (p = 0.05). CONCLUSION: Targeted sonography detected nearly half (46%) of MRI lesions evaluated, with depiction being most frequent for masses. Sonographically occult lesions had a probability of malignancy (22%), which warrants biopsy despite lack of sonographic detection. PMID- 19304725 TI - Atypical ductal hyperplasia diagnosed at sonographically guided 14-gauge core needle biopsy of breast mass. AB - OBJECTIVE: The purpose of this study was to evaluate the surgical outcome of atypical ductal hyperplasia (ADH) diagnosed at sonographically guided 14-gauge core needle biopsy of breast masses and to determine whether the clinical, procedural, and radiologic features of this lesion can be used to predict upgrade to malignancy. MATERIALS AND METHODS: We retrospectively reviewed the pathologic results of sonographically guided 14-gauge core needle biopsy of solid breast masses. A total of 21 ADH lesions diagnosed with this procedure and surgically excised were included in the study. For each lesion, medical records and images were reviewed, and clinical, procedural, and radiologic variables were coded. The mammographic and sonographic features were determined, and the BI-RADS assessment category was noted. We compared underestimation rates among the collected variables. RESULTS: The results of surgical excision of 21 ADH lesions were malignancy in 13 cases (62% rate of underestimation of ADH). In comparison of rates of underestimation of malignancy among clinical and procedural variables, no statistically significant differences were found. There also was no significant difference among lesion characteristics and BI-RADS categories at mammography and sonography. CONCLUSION: ADH diagnosed at sonographically guided 14-gauge core needle biopsy has a high underestimation rate with respect to the results of surgical excision. Surgical excision should be recommended when ADH is diagnosed at sonographically guided 14-gauge core needle biopsy of breast masses. PMID- 19304726 TI - Increasing accuracy of detection of breast cancer with 3-T MRI. AB - OBJECTIVE: The purposes of this study were to assess the sensitivity and specificity of 3-T MRI compared with those of mammography and sonography in the evaluation of breast cancer and to evaluate the results in light of previously published data obtained with 1-T and 1.5-T systems. MATERIALS AND METHODS: A retrospective review was conducted with data on 434 women (mean age, 53 years) who underwent MRI evaluation of the breasts (n = 868). Results obtained from 3-T MRI, mammographic, and sonographic examinations of all patients were compared. Sensitivity, specificity, and predictive values were calculated, and statistics were analyzed with the chi-square test. RESULTS: The sensitivities of MRI, mammography, and sonography in the detection of malignancy were 100%, 81.8%, and 86.4%, and the specificities were 93.9%, 99%, and 98.1%. The specificity of MRI increased from 92.8% to 94.5% over the course of the study (2006-2007). Compared with mammography (p < 0.001) and sonography (p = 0.001), MRI depicted a significantly higher number of malignant tumors of the breast. There was no significant difference between mammography and sonography (p = 0.095). Results were compared with those in earlier reports in the literature. CONCLUSION: MRI at 3 T is more sensitive than mammography and sonography in the detection of breast cancer and the characterization of small lesions (reaching 4 mm) but has lower specificity. Compared with the results of 1-T and 1.5-T MRI in the literature, 3 T MRI has higher sensitivity in the detection of breast cancer with no significant difference in specificity. PMID- 19304727 TI - Patients' perceptions of breast MRI: a single-center study. AB - OBJECTIVE: The high sensitivity and lower specificity of breast MRI for detecting breast cancer are well documented in the literature. As the use of breast MRI continues to grow, medical controversy persists over the appropriate use of this test. However, patients' perceptions of breast MRI are unknown. The aim of this study was to determine patients' perception of the impact of breast MRI on their care and to compare patients' perception with objective assessments. MATERIALS AND METHODS: Patients undergoing breast MRI between January 2004 and December 2005 for cancer staging or high-risk surveillance were surveyed regarding comfort during, satisfaction with, and perception of MRI's impact on their clinical care. The respondents' perceptions of the impact of MRI on their care were compared with objective assessments based on retrospective chart review for breast cancer patients. RESULTS: Of 300 patients, the survey was completed by 227 (76%). Ninety one percent (203/224) reported mild or no discomfort during the examination, and 91% (206/226) perceived the breast MRI examination as having a positive impact on their care. Breast cancer patients were significantly more likely to perceive the impact of MRI as beneficial in comparison with the objective assessment (91% vs 35%, respectively; p < 0.001). CONCLUSION: Most women undergoing breast MRI for cancer staging and screening found it to be a comfortable test and perceived it to have a positive impact on their care. Breast cancer patients perceived the clinical impact of their breast MRI examination to be significantly more positive than when assessed objectively. PMID- 19304728 TI - Normal responses to restraint stress in mice lacking the gene for neuronal nitric oxide synthase. AB - The hormonal changes associated with immobilization stress (IMO) include a swift increase in corticosterone (CORT) concentration and a decrease in circulating testosterone (T) levels. There is evidence that the production of the short-lived neuromodulator nitric oxide (NO) is increased during stress in various tissues, including the brain. NO also suppresses the biosynthesis of T. Both the inducible and the neuronal isoforms of NO synthase (iNOS and nNOS, respectively) have been implicated in this suppression, but the evidence has not been conclusive. We used adult wild-type (WT) and nNOS knockout male mice (nNOS-/-) to assess the respective roles of CORT and nNOS-derived NO in stress mediated inhibition of T production. Animals were assigned to either basal control or 3-hour IMO groups. No difference in basal plasma and testicular T levels were observed between WT and nNOS-/-, although testicular weights of mutant mice were slightly lower compared to WT animals. The plasma contents of luteinizing hormone (LH) and CORT in unstressed mice of both genotypes were similar. Exposure to 3 hours of IMO increased plasma CORT and decreased T concentrations in mice of both genotypes. However, comparable levels of plasma LH and testicular nitrite and nitrate (NOx), NO stable metabolites, were detected in control and stressed WT and nNOS-/- mice. Adrenal concentrations of NOx declined after IMO, but the reduction was not statistically significant. These findings implicate CORT rather than NO generated by nNOS in the rapid stress-induced suppression of circulating T. PMID- 19304729 TI - Impact of health reforms on child health services in Europe: the case of Bulgaria. AB - BACKGROUND: In the last two decades, all countries in Europe have embarked on substantial health reforms, introducing new models of financing and provision of health services. Using Bulgaria as a case study, this article examines the impact of the reforms on child health services. METHODS: This is the first of a series of papers drawing on a broader research on inequalities in access to child health services, using Bulgaria as a case study. Multiple methods and data sources were used, including a review of the literature and existing epidemiological data, 50 qualitative in-depth interviews and an analysis of regulatory documents. This article presents the findings of the documentary analysis. RESULTS: Primary health services for children are now provided by general practitioners. Children are exempted from health insurance contributions and user fees and are formally entitled to free health care. During the first years of the reform general practitioners still had insufficient training in child health. Restrictions on the number of referrals to paediatricians and discontinuation of community services at a time when general practice was not well established, undermined access to quality care. CONCLUSION: While many of these issues have been subsequently addressed, the reform process was far from linear. Challenges remain in ensuring access to quality child health services to the rural population and marginalized groups, such as the Roma minority and children with disabilities. Throughout Europe, health reforms need to be based on solid evidence of what works best for improving quality of and access to child health services. PMID- 19304730 TI - The gap between suicide characteristics in the print media and in the population. AB - BACKGROUND: Programmes to educate media professionals about suicide are increasingly established, but information about which suicide cases are most likely to be reported in the mass media is sparse. METHODS: We applied binomial tests to compare frequencies of social characteristics of all domestic suicides in the 13 largest Austrian print media in 2005 with frequencies of suicide characteristics in the population. Additionally, each reported suicide case was linked to its respective entry in the suicide database. We performed a logistic regression analysis, with presence of an article as outcome, and sex of the suicide case, age, religious affiliation, family status, conduction of an autopsy and location of the suicide as explaining variables. Time of the year and federal state where the suicide happened was controlled for. RESULTS: Binomial tests showed that suicides involving murder or murder attempt were over-represented in the media. Reporting on mental disorders was under-represented. In the regression analysis, the likelihood of a report was negatively associated with the age of suicide cases. Foreign citizenship was a further predictor of a suicide report. The methods of drowning, jumping, shooting and rare methods were more likely to be reported than hanging, which is the most frequent suicide method in Austria. CONCLUSIONS: Suicide characteristics in the media are not representative of the population. The identified discrepancies provide a basis for tailor-made education of mass media professionals. PMID- 19304731 TI - Self-reporting weight and height: misclassification effect on the risk estimates for acute myocardial infarction. AB - BACKGROUND: The accuracy of self-reported weight and height to measure obesity has been evaluated, but no information is available on the possible error effects of self-reporting when estimating the association between body mass index (BMI) categories and the occurrence of acute myocardial infarction (AMI). We aim to evaluate if two different sources of information on height and weight (reported vs. measured) result in different risk estimates for non-fatal AMI events. METHODS: A population-based case-control study was conducted with 732 cases of first AMI and 1914 community controls, recruited from the same catchment area of hospitals. As part of an interview, participants self-reported weight and height immediately before having it measured. Data were analysed separately by sex and age strata (< or =45; >45 years). RESULTS: Women under-reported their weight and over-reported their height, and the mean differences between measured and self reported data were significantly larger in controls. Male controls also under reported their weight, but cases over-reported it. After adjustment, in younger women the use of self-reported data underestimated the AMI risk according to BMI categories, but in older ones the self-reporting overestimated AMI risk, although with no statistical significance. In younger men, the association between AMI and self-reported obesity (BMI > or = 30 kg/m(2)) was overestimated in approximately 50% (measured: OR = 2.05, 95% CI 1.08-3.87; self-reported: OR = 3.06, 95% CI 1.56 6.00). In older participants, a significant association was only found for overweight men when using self-reported data. CONCLUSIONS: Self-reporting of height and weight produced a differential misclassification and biased risks for AMI according to BMI, affecting not only the magnitude, but also the estimates direction. PMID- 19304732 TI - Psychosomatic health problems among adolescents in Sweden--are the time trends gender related? AB - BACKGROUND: Since the economic recession in Sweden in the 1990s alarming reports about deteriorating mental and psychosomatic health among young people have repeatedly been published but reliable survey data are rare. The purpose of the study is to describe the trends in psychosomatic health problems among adolescents, focusing on gender differences. METHODS: The analysis is based on repeated cross-sectional data collected 1988-2005 among about 15,000 adolescents (15- to 16-years old) within a county in Sweden. The data were collected in schools using a questionnaire that was completed anonymously. A composite measure of psychosomatic health problems based on eight items was used. RESULTS: Psychosomatic health complaints among boys increased mainly during the in recession time period, while the health problems among girls increased only slightly during the crisis, but increased dramatically in the post-recession time period. Only among boys did the variance in psychosomatic health increase successively across years of investigation, implying that the psychosomatic health among boys on average did not change over time. CONCLUSIONS: In showing different trend patterns across genders, the present study nuances and qualifies previous reports on deteriorating mental and psychosomatic health among adolescents. Whether the gender-related trend patterns are due to differences in the relative influence of economic and social stress factors or if they reflect other factors such as changes in the educational systems should be addressed in future studies. PMID- 19304733 TI - Daptomycin pharmacodynamics against Staphylococcus aureus hemB mutants displaying the small colony variant phenotype. AB - OBJECTIVES: Staphylococcus aureus small colony variants (SCVs) are slow-growing morphological variants associated with persistent infections. While vancomycin activity has been shown to be attenuated against SCVs of S. aureus, few data exist regarding daptomycin. The objective was to evaluate the pharmacodynamics of daptomycin against defined S. aureus mutants displaying the SCV phenotype. METHODS: Two S. aureus hemB mutants (Ia48 and III33) displaying the SCV phenotype and their parental strains (COL and Newman) were evaluated. Time-kill experiments were performed using a starting inoculum of 10(6) cfu/mL at 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32 and 64 times the MIC. Samples were obtained at 0, 1, 2, 4, 6, 8 and 24 h, plated and incubated to determine colony counts. A Hill-type pharmacodynamic mathematical model was fitted to the data to characterize the effect. RESULTS: Bactericidal activity for daptomycin was achieved and occurred in a concentration-dependent manner against both hemB mutants and their parental strains. Against strains with normal phenotype, bactericidal activity was achieved rapidly, within 2 h at concentrations > or =16 times the MIC, while against SCVs, bactericidal activity was achieved within 6 h at concentrations > or =16 times the MIC. Against both hemB mutants, daptomycin maintained bactericidal activity at 24 h, with similar profiles of killing activity when compared with their parental strains. CONCLUSIONS: Daptomycin achieved bactericidal activity against S. aureus hemB mutants and parenteral isolates. Daptomycin represents a potential therapeutic option for infections caused by S. aureus strains displaying the SCV phenotype and additional studies are warranted. PMID- 19304735 TI - Wellness appraisal among adolescents in Jordan: a model from a developing country: a cross-sectional questionnaire survey. AB - The demographic and economic transition that many developing countries, including Jordan, are undergoing is producing important changes in diet and lifestyle that greatly impact the development of chronic illness. The health behavior of adolescents in developing countries constitutes one of the most serious global challenges we face. The purpose of this study was to explore the wellness appraisal of Jordanian adolescents. It specifically describes the (i) self-care and health history pattern, (ii) to assess physical activity and nutrition appraisal, (iii) quality of life appraisal and (iv) school and outside activities appraisal. A self-administrated questionnaire collected the data from adolescent groups (boys and girls.). A multi-stage stratified random sample was obtained from six public schools by first selecting the educational directorate located in the city of Irbid, which is located in the north of Jordan. A random sub sample of six individual schools was then selected. Individual classes were then selected. A total of 269 boys and 261 girls (12-17-year old) were included in the analysis. The results showed that the students engaged in risky health behaviors which could lead to short- and long-term health problems. The most alarming finding of this study was students' nutritional habits, including less than the daily requirements of fruits, vegetables, milk and meat, while the intake of fast food, soft drinks and sweets were higher than recommended. School-based health promotion and wellness programs should be established in Jordan to influence the health behaviors of adolescents and parents and to avoid further deterioration of their health. Jordanian school curriculum needs to integrate more precise health education programs about diet, exercise, self care and other life style behaviors. More detailed studies are needed with more elaborate instruments about food habits, physical activities and psycho social life. PMID- 19304734 TI - K(ATP) activation prevents progression of cardiac hypertrophy to failure induced by pressure overload via protecting endothelial function. AB - AIMS: We investigated the effects of iptakalim, a new ATP-sensitive potassium channel (K(ATP)) opener providing endothelial protection, on the progression of cardiac hypertrophy to failure in a rat model of pressure overloading caused by abdominal aortic banding (AAB). Endothelial dysfunction is central to cardiac hypertrophy and failure induced by pressure overload. It would be useful to clarify whether iptakalim could prevent this. METHODS AND RESULTS: The effects of pressure overload were assessed in male Sprague-Dawley rats 6 weeks after AAB using progression of cardiac hypertrophy to heart failure as the endpoint. The AAB-treated rats had significantly elevated blood pressure, systolic and diastolic cardiac dysfunction, evidence of left ventricular hypertrophy (LVH), and transition to heart failure. LVH was characterized by increases in the ratios of heart and left ventricular weights to body weight, increased myocyte cross sectional areas, myocardial and perivascular fibrosis, and elevated cardiac hydroxyproline. These could be prevented by treatment with iptakalim at daily oral doses of 1, 3, and 9 mg/kg for 6 weeks. Progression to cardiac failure, demonstrated by increases in relative lung and right ventricular weights, cardiac function disorders and overexpression of atrial and B-type natriuretic peptide mRNA, could also be prevented. The downregulated nitric oxide signalling system was enhanced, whereas the upregulated endothelin signalling system was inhibited, resulting in normalization of the balance between these two systems. CONCLUSION: Iptakalim protected the endothelium and prevented progression of cardiac hypertrophy to failure induced by a pressure overload. PMID- 19304736 TI - A comparative study on resilience level between WHO health promoting schools and other schools among a Chinese population. AB - The WHO health promoting school (HPS) approach covers key areas including school based programmes improving students' psychological health, but there have been few studies evaluating the resilience performance of these schools. This study compared the resilience scores between schools within the healthy school award (HSA) scheme (HPS group) and those not (non-HPS group). We conducted a cross sectional survey of grade-one students (aged 12), all teachers and parents of mainstream secondary schools recruited by stratified random sampling in one large Territory of Hong Kong using validated resilience questionnaires during November December 2005. Four non-HPS and four HPS secondary schools were recruited, respectively, involving 1408 students, 891 parents and 91 teachers, with similar baseline characteristics. The HPS students were found to have better scores than non-HPS students (average age 12.4 year-old in both groups) in all dimensions with significantly higher scores in 'Peer Support' (p = 0.013), 'Making a Difference' (p = 0.011), 'About Me' (p = 0.027) and 'Generally Happy' (p = 0.011). There was no difference in the scores between non-HPS and HPS parents. The HPS teachers reported significantly higher scores in 'Health Policies' (p = 0.023), 'Social Environment' (p = 0.049), 'School Community Relations' (p = 0.048), 'Personal Skills Building' (p = 0.008) and 'Partnership & Health Services' (p = 0.047). The secondary HPS students and teachers reported significantly higher resilience scores than those of non-HPS. This study shows that the HSA scheme under WHO has the potential to exert positive changes in students and teachers and the concept of HPS is effective in building resilience among major school stakeholders. PMID- 19304737 TI - The historical origins of the basic concepts of health promotion and education: the role of ancient Greek philosophy and medicine. AB - Although it is commonly accepted that the basic concepts of 'Health Promotion' have been developed in the last two decades, they have their roots in ancient civilizations and in particular in Greek antiquity. As evident from medical and philosophical documents of the sixth to fourth centuries B.C., the ancient Greeks were the first to break with the supernatural conceptions of health and disease that had so far dominated human societies. The ancient Greeks developed the physiocratic school of thought, realizing that maintaining good health and fighting illness depend on natural causes and that health and disease cannot be dissociated from particular physical and social environments nor from human behavior. In this context, they defined health as a state of dynamic equilibrium between the internal and the external environment, they took under consideration the physical and social determinants of health, they empowered individuals and communities through new democratic and participatory institutions, they gave emphasis in health education and skill development, they recognized the importance of supportive environments and of healthy public policy and they re oriented medicine toward a more naturalistic and humanistic perspective. The aim of the present study is to highlight such core concepts from these early times that helped establishing the foundations for health promotion and education in the modern era according to the Ottawa Charter. PMID- 19304738 TI - Responses to desiccation stress in lichens are different from those in their photobionts. AB - In order to clarify the role of symbiotic association in desiccation tolerance of photosynthetic partners in lichens, responses to air-drying and hypertonic treatments in a green-algal lichen (a chlorolichen, Ramalina yasudae Rasanen) and its green algal photobiont (freshly released and cultured Trebouxia sp.) were studied. Responses to dehydration in the isolated Trebouxia sp. were different from those in the lichen, R. yasudae, i.e. (i) the PSII reaction was totally inhibited in R. yasudae when photosynthesis was completely inhibited by desiccation, but it remained partially active in isolated Trebouxia sp; (ii) dehydration-induced quenching of PSII fluorescence was less in the isolated Trebouxia sp. compared with that in R. yasudae, suggesting that a substance(s) or a mechanism(s) to dissipate absorbed light energy to heat was lost by the isolation of the photobiont; and (iii) the air-dried isolated Trebouxia sp. showed a higher sensitivity to photoinhibition than R. yasudae. These results support the idea that association of the photobionts with the mycobionts increases tolerance to photoinhibition under drying conditions. PMID- 19304739 TI - Enhanced defense responses in Arabidopsis induced by the cell wall protein fractions from Pythium oligandrum require SGT1, RAR1, NPR1 and JAR1. AB - The cell wall protein fraction (CWP) is purified from the non-pathogenic biocontrol agent Pythium oligandrum and is composed of two glycoproteins (POD-1 and POD-2), which are structurally similar to class III elicitins. In tomato plants treated with CWP, jasmonic acid (JA)- and ethylene (ET)-dependent signaling pathways are activated, and resistance to Ralstonia solanaceraum is enhanced. To dissect CWP-induced defense mechanisms, we investigated defense gene expression and resistance to bacterial pathogens in Arabidopsis thaliana ecotype Col-0 treated with CWP. When the leaves of Col-0 were infiltrated with CWP, neither visible necrosis nor salicylic acid (SA)-responsive gene (PR-1 and PR-5) expression was induced. In contrast, JA-responsive gene (PDF1.2 and JR2) expression was up-regulated and the resistance to R. solanaceraum and Pseudomonas syringae pv. tomato DC3000 was enhanced in response to CWP. Such CWP-induced defense responses were completely compromised in CWP-treated coi1-1 and jar1-1 mutants with an impaired JA signaling pathway. The induction of defense-related gene expression after CWP treatment was partially compromised in ET-insensitive ein2-1 mutants, but not in SA signaling mutants or nahG transgenic plants. Global gene expression analysis using cDNA array also suggested that several other JA- and ET-responsive genes, but not SA-responsive genes, were up-regulated in response to CWP. Further analysis of CWP-induced defense responses using another eight mutants with impaired defense signaling pathways indicated that, interestingly, the induction of JA-responsive gene expression and enhanced resistance to two bacterial pathogens in response to CWP were completely compromised in rar1-1, rar1-21, sgt1a-1, sgt1b (edm1) and npr1-1 mutants. Thus, the CWP-induced defense system appears to be regulated by JA-mediated and SGT1-, RAR1- and NPR1-dependent signaling pathways. PMID- 19304740 TI - The candidate gene XIRP2 at a quantitative gene locus on equine chromosome 18 associated with osteochondrosis in fetlock and hock joints of South German Coldblood horses. AB - A whole-genome scan for radiological signs of osteochondrosis (OC) and osteochondrosis dissecans (OCD) in South German Coldblood (SGC) horses using 250 microsatellite markers identified a genome-wide significant quantitative trait locus (QTL) for fetlock OCD and a chromosome-wide QTL for hock OC on Equus caballus chromosome (ECA) 18 at a relative position of 45.9-78.2 cM. The aim of this study was to analyze associations of single-nucleotide polymorphisms (SNPs) in candidate genes for OC in this QTL region using 96 SGC horses. The OC-QTL on ECA18 could be confirmed and narrowed down to an interval of 13 Mb between GALNT13 and Xin actin-binding repeat containing 2 (XIRP2). SNPs in the XIRP2 gene were significantly associated with fetlock OC, fetlock OCD, and hock OC. The significant associations of SNPs in XIRP2 could be confirmed in linear animal models controlling for systematic environmental and residual quantitative genetic effects. The significant additive genetic effects of the intronic SNPs (AJ885515:g.159A>G, AJ885515:g.445T>C) in XIRP2 were 0.15 (P = 0.01) for fetlock OC, 0.27 (P = 0.01) for fetlock OCD, and 0.15-0.16 (P = 0.01-0.02) for hock OC. Homozygous (A/A or T/T) and heterozygous horses were at a 1.3- to 2.4-fold higher risk for fetlock and hock OC. These results suggest that dominant variants of XIRP2 may be involved in pathogenesis of equine OC. PMID- 19304741 TI - Species delineation and evolutionary history of the globally distributed spotted eagle ray (Aetobatus narinari). AB - The spotted eagle ray (Aetobatus narinari), a large coral reef-associated batoid of conservation concern, is currently described as a single, circumglobally distributed species. However, geographic differences in its morphology and parasite diversity have raised unconfirmed suspicions that A. narinari may constitute a species complex. We used 1570 bp of mitochondrial and nuclear sequence data (cytochrome b, cytochrome c oxidase subunit I, and internal transcribed spacer 2) to assess the validity of A. narinari as a single cosmopolitan species and infer its evolutionary history. Specimens from 4 major geographic regions were examined: the Central Atlantic, Eastern Pacific, Western Pacific, and Central Pacific. Phylogenies described 3 distinct, reciprocally monophyletic lineages with no genetic exchange among regions. Based on combined genealogical concordance and genetic distance criteria, we recommend that the Western/Central Pacific lineage be recognized as a distinct species from lineages in the Central Atlantic and Eastern Pacific. The latter 2 lineages, separated by the Isthmus of Panama, are proposed as subspecies. A basal position in phylogenetic analyses and statistical parsimony results support an Indo-West Pacific origin for the A. narinari species complex, with subsequent westerly dispersal around the southern tip of Africa into the Atlantic and then into the Eastern Pacific. PMID- 19304742 TI - Impact of left ventricular volume/mass ratio on diastolic function. AB - AIMS: To assess the impact of left ventricular (LV) volume/mass ratio on diastolic function parameters in subjects with dilated cardiomyopathy (DCM) or hypertrophic cardiomyopathy (HCM) and healthy controls. METHODS AND RESULTS: We performed echocardiography in 44 healthy controls, 35 HCM subjects, 29 DCM subjects with narrow QRS complex (DCM-n), and 27 DCM subjects with wide QRS complex (DCM-w). Mitral annulus velocity (E(a)) and transmitral E-wave velocity were used to estimate time constant of isovolumic pressure decay (tau). LV flow propagation velocity (V(p)) and early intraventricular pressure gradient (IVPG) were derived from colour M-mode of LV inflow. We calculated LV twist and peak untwisting rate (UntwR) by speckle tracking. Mean LV volume/mass ratio was 0.34 +/- 0.09 mL/g in healthy controls, 0.15 +/- 0.06 mL/g in HCM, 0.6 +/- 0.2 mL/g in DCM-n, and 0.8 +/- 0.3 mL/g in DCM-w patients (P < 0.001 for all groups). Resting LV ejection fractions were 63 +/- 7, 64 +/- 8, 31 +/- 8, and 26 +/- 8%, respectively (P < 0.01 vs. controls for DCM groups). In a multivariate analysis, LV volume/mass ratio remained a strong independent predictor of V(p) (P < 0.001), IVPG (P = 0.009), and UntwR (P < 0.001) but not for E(a) (P = 0.25). CONCLUSION: LV volume/mass ratio had influences on diastolic function parameters independent of intrinsic diastolic function and filling pressures. It should be considered when assessing patients suspected of LV diastolic dysfunction. PMID- 19304743 TI - p53 as a specific prognostic factor in triple-negative breast cancer. AB - OBJECTIVE: A recent suggestion is that the predictive value of a single biomarker may rely on the genetic background on the tumor and that different breast cancer subgroups may have different predictive markers of response to chemotherapy. The prognostic value of p53 in the outcome of adjuvant anthracycline-containing chemotherapy was evaluated according to molecular subclasses defined using the expression of estrogen receptor, progesterone receptor and human epidermal growth factor receptor 2. METHODS: Subjects were patients (n = 135) with invasive ductal carcinoma treated with adjuvant anthracycline-based chemotherapy between 1994 and 2000 in our hospital. Clinico-pathological features were reviewed by retrospective examination of medical records. RESULTS: Overall survival rate was not independently predictive by p53 status (P = 0.182). However, in triple negative cases, there was statistically significant survival difference (P = 0.034) and no statistically significant difference (P = 0.783) in non-triple negative cases by p53 status. In the Cox proportional hazard analysis, p53 was also strongly predictive for relapse-free survival (P = 0.013) and overall survival (P = 0.049) in triple-negative patients. CONCLUSIONS: p53 status could be a specific prognostic factor in triple-negative breast cancer patients treated by adjuvant anthracycline-based regimen. When p53 is positive in triple-negative breast cancer, we could expect poor survival, prompting aggressive or alternative treatment. PMID- 19304744 TI - A role for hydrophobicity in a Diels-Alder reaction catalyzed by pyridyl-modified RNA. AB - New classes of RNA enzymes or ribozymes have been obtained by in vitro evolution and selection of RNA molecules. Incorporation of modified nucleotides into the RNA sequence has been proposed to enhance function. DA22 is a modified RNA containing 5-(4-pyridylmethyl) carboxamide uridines, which has been selected for its ability to promote a Diels-Alder cycloaddition reaction. Here, we show that DA_TR96, the most active member of the DA22 RNA sequence family, which was selected with pyridyl-modified nucleotides, accelerates a cycloaddition reaction between anthracene and maleimide derivatives with high turnover. These widely used reactants were not used in the original selection for DA22 and yet here they provide the first demonstration of DA_TR96 as a true multiple-turnover catalyst. In addition, the absence of a structural or essential kinetic role for Cu(2+), as initially postulated, and nonsequence-specific hydrophobic interactions with the anthracene substrate have led to a reevaluation of the pyridine modification's role. These findings broaden the catalytic repertoire of the DA22 family of pyridyl-modified RNAs and suggest a key role for the hydrophobic effect in the catalytic mechanism. PMID- 19304745 TI - Structural insights into the cooperative binding of SeqA to a tandem GATC repeat. AB - SeqA is a negative regulator of DNA replication in Escherichia coli and related bacteria that functions by sequestering the origin of replication and facilitating its resetting after every initiation event. Inactivation of the seqA gene leads to unsynchronized rounds of replication, abnormal localization of nucleoids and increased negative superhelicity. Excess SeqA also disrupts replication synchrony and affects cell division. SeqA exerts its functions by binding clusters of transiently hemimethylated GATC sequences generated during replication. However, the molecular mechanisms that trigger formation and disassembly of such complex are unclear. We present here the crystal structure of a dimeric mutant of SeqA [SeqADelta(41-59)-A25R] bound to tandem hemimethylated GATC sites. The structure delineates how SeqA forms a high-affinity complex with DNA and it suggests why SeqA only recognizes GATC sites at certain spacings. The SeqA-DNA complex also unveils additional protein-protein interaction surfaces that mediate the formation of higher ordered complexes upon binding to newly replicated DNA. Based on this data, we propose a model describing how SeqA interacts with newly replicated DNA within the origin of replication and at the replication forks. PMID- 19304746 TI - Structural analysis and DNA binding of the HMG domains of the human mitochondrial transcription factor A. AB - The mitochondrial transcription factor A (mtTFA) is central to assembly and initiation of the mitochondrial transcription complex. Human mtTFA (h-mtTFA) is a dual high mobility group box (HMGB) protein that binds site-specifically to the mitochondrial genome and demarcates the promoters for recruitment of h-mtTFB1, h mtTFB2 and the mitochondrial RNA polymerase. The stoichiometry of h-mtTFA was found to be a monomer in the absence of DNA, whereas it formed a dimer in the complex with the light strand promoter (LSP) DNA. Each of the HMG boxes and the C terminal tail were evaluated for their ability to bind to the LSP DNA. Removal of the C-terminal tail only slightly decreased nonsequence specific DNA binding, and box A, but not box B, was capable of binding to the LSP DNA. The X-ray crystal structure of h-mtTFA box B, at 1.35 A resolution, revealed the features of a noncanonical HMG box. Interactions of box B with other regions of h-mtTFA were observed. Together, these results provide an explanation for the unusual DNA binding properties of box B and suggest possible roles for this domain in transcription complex assembly. PMID- 19304747 TI - Lesion-induced DNA weak structural changes detected by pulsed EPR spectroscopy combined with site-directed spin labelling. AB - Double electron-electron resonance (DEER) was applied to determine nanometre spin spin distances on DNA duplexes that contain selected structural alterations. The present approach to evaluate the structural features of DNA damages is thus related to the interspin distance changes, as well as to the flexibility of the overall structure deduced from the distance distribution. A set of site-directed nitroxide-labelled double-stranded DNA fragments containing defined lesions, namely an 8-oxoguanine, an abasic site or abasic site analogues, a nick, a gap and a bulge structure were prepared and then analysed by the DEER spectroscopic technique. New insights into the application of 4-pulse DEER sequence are also provided, in particular with respect to the spin probes' positions and the rigidity of selected systems. The lesion-induced conformational changes observed, which were supported by molecular dynamics studies, confirm the results obtained by other, more conventional, spectroscopic techniques. Thus, the experimental approaches described herein provide an efficient method for probing lesion induced structural changes of nucleic acids. PMID- 19304748 TI - Flow cytometry for enrichment and titration in massively parallel DNA sequencing. AB - Massively parallel DNA sequencing is revolutionizing genomics research throughout the life sciences. However, the reagent costs and labor requirements in current sequencing protocols are still substantial, although improvements are continuously being made. Here, we demonstrate an effective alternative to existing sample titration protocols for the Roche/454 system using Fluorescence Activated Cell Sorting (FACS) technology to determine the optimal DNA-to-bead ratio prior to large-scale sequencing. Our method, which eliminates the need for the costly pilot sequencing of samples during titration is capable of rapidly providing accurate DNA-to-bead ratios that are not biased by the quantification and sedimentation steps included in current protocols. Moreover, we demonstrate that FACS sorting can be readily used to highly enrich fractions of beads carrying template DNA, with near total elimination of empty beads and no downstream sacrifice of DNA sequencing quality. Automated enrichment by FACS is a simple approach to obtain pure samples for bead-based sequencing systems, and offers an efficient, low-cost alternative to current enrichment protocols. PMID- 19304749 TI - Gene structures and processing of Arabidopsis thaliana HYL1-dependent pri-miRNAs. AB - Arabidopsis thaliana HYL1 is a nuclear double-stranded RNA-binding protein involved in the maturation of pri-miRNAs. A quantitative real-time PCR platform for parallel quantification of 176 pri-miRNAs was used to reveal strong accumulation of 57 miRNA precursors in the hyl1 mutant that completely lacks HYL1 protein. This approach enabled us for the first time to pinpoint particular members of MIRNA family genes that require HYL1 activity for efficient maturation of their precursors. Moreover, the accumulation of miRNA precursors in the hyl1 mutant gave us the opportunity to carry out 3' and 5' RACE experiments which revealed that some of these precursors are of unexpected length. The alignment of HYL1-dependent miRNA precursors to A. thaliana genomic sequences indicated the presence of introns in 12 out of 20 genes studied. Some of the characterized intron-containing pri-miRNAs undergo alternative splicing such as exon skipping or usage of alternative 5' splice sites suggesting that this process plays a role in the regulation of miRNA biogenesis. In the hyl1 mutant intron-containing pri miRNAs accumulate alongside spliced pri-miRNAs suggesting the recruitment of HYL1 into the miRNA precursor maturation pathway before their splicing occurs. PMID- 19304750 TI - The proapoptotic dp5 gene is a direct target of the MLK-JNK-c-Jun pathway in sympathetic neurons. AB - The death of sympathetic neurons after nerve growth factor (NGF) withdrawal requires de novo gene expression. Dp5 was one of the first NGF withdrawal-induced genes to be identified and it encodes a proapoptotic BH3-only member of the Bcl-2 family. To study how dp5 transcription is regulated by NGF withdrawal we cloned the regulatory regions of the rat dp5 gene and constructed a series of dp5 luciferase reporter plasmids. In microinjection experiments with sympathetic neurons we found that three regions of dp5 contribute to its induction after NGF withdrawal: the promoter, a conserved region in the single intron, and sequences in the 3' untranslated region of the dp5 mRNA. A construct containing all three regions is efficiently activated by NGF withdrawal and, like the endogenous dp5, its induction requires mixed-lineage kinase (MLK) and c-Jun N-terminal kinase (JNK) activity. JNKs phosphorylate the AP-1 transcription factor c-Jun, and thereby increase its activity. We identified a conserved ATF site in the dp5 promoter that binds c-Jun and ATF2, which is critical for dp5 promoter induction after NGF withdrawal. These results suggest that part of the mechanism by which the MLK-JNK-c-Jun pathway promotes neuronal apoptosis is by activating the transcription of the dp5 gene. PMID- 19304751 TI - Selection of aptamers for a protein target in cell lysate and their application to protein purification. AB - Functional genomics requires structural and functional studies of a large number of proteins. While the production of proteins through over-expression in cultured cells is a relatively routine procedure, the subsequent protein purification from the cell lysate often represents a significant challenge. The most direct way of protein purification from a cell lysate is affinity purification using an affinity probe to the target protein. It is extremely difficult to develop antibodies, classical affinity probes, for a protein in the cell lysate; their development requires a pure protein. Thus, isolating the protein from the cell lysate requires antibodies, while developing antibodies requires a pure protein. Here we resolve this loop problem. We introduce AptaPIC, Aptamer-facilitated Protein Isolation from Cells, a technology that integrates (i) the development of aptamers for a protein in cell lysate and (ii) the utilization of the developed aptamers for protein isolation from the cell lysate. Using MutS protein as a target, we demonstrate that this technology is applicable to the target protein being at an expression level as low as 0.8% of the total protein in the lysate. AptaPIC has the potential to considerably speed up the purification of proteins and, thus, accelerate their structural and functional studies. PMID- 19304752 TI - From damaged genome to cell surface: transcriptome changes during bacterial cell death triggered by loss of a restriction-modification gene complex. AB - Genetically programmed cell deaths play important roles in unicellular prokaryotes. In postsegregational killing, loss of a gene complex from a cell leads to its descendants' deaths. With type II restriction-modification gene complexes, such death is triggered by restriction endonuclease's attacks on under methylated chromosomes. Here, we examined how the Escherichia coli transcriptome changes after loss of PaeR7I gene complex. At earlier time points, activation of SOS genes and sigma(E)-regulon was noticeable. With time, more SOS genes, stress response genes (including sigma(S)-regulon, osmotic-, oxidative- and periplasmic stress genes), biofilm-related genes, and many hitherto uncharacterized genes were induced, and genes for energy metabolism, motility and outer membrane biogenesis were repressed. As expected from the activation of sigma(E)-regulon, the death was accompanied by cell lysis and release of cellular proteins. Expression of several sigma(E)-regulon genes indeed led to cell lysis. We hypothesize that some signal was transduced, among multiple genes involved, from the damaged genome to the cell surface and led to its disintegration. These results are discussed in comparison with other forms of programmed deaths in bacteria and eukaryotes. PMID- 19304753 TI - Promoter targeted small RNAs induce long-term transcriptional gene silencing in human cells. AB - Small RNAs targeted to gene promoters in human cells can mediate transcriptional gene silencing (TGS) by directing silent state epigenetic modifications to targeted loci. Many mechanistic details of this process remain poorly defined, and the ability to stably modulate gene expression in this manner has not been explored. Here we describe the mechanisms of establishment and maintenance of long-term transcriptional silencing of the human ubiquitin C gene (UbC). Sustained targeting of the UbC promoter with a small RNA for a minimum of 3 days resulted in long-term silencing which correlated with an early increase in histone methylation and a later increase in DNA methylation at the targeted locus. Transcriptional silencing of UbC required the presence of a promoter associated RNA. The establishment and maintenance of the TGS were shown to require distinct protein factors. Argonaute 1 (Ago1), DNA methyltransferase 3a (DNMT3a) and histone deacetylase 1 (HDAC1) were required for the initiation of silencing, and DNA methyltransferase 1 (DNMT1) was necessary for maintenance. Taken together the data presented here highlight the cellular pathway with which noncoding RNAs interact to epigenetically regulate gene expression in human cells. PMID- 19304754 TI - The transcriptional coactivator MAML1 regulates p300 autoacetylation and HAT activity. AB - MAML1 is a transcriptional coregulator originally identified as a Notch coactivator. MAML1 is also reported to interact with other coregulator proteins, such as CDK8 and p300, to modulate the activity of Notch. We, and others, previously showed that MAML1 recruits p300 to Notch-regulated genes through direct interactions with the DNA-CSL-Notch complex and p300. MAML1 interacts with the C/H3 domain of p300, and the p300-MAML1 complex specifically acetylates lysines of histone H3 and H4 tails in chromatin in vitro. In this report, we show that MAML1 potentiates p300 autoacetylation and p300 transcriptional activation. MAML1 directly enhances p300 HAT activity, and this coincides with the translocation of MAML1, p300 and acetylated histones to nuclear bodies. PMID- 19304755 TI - Creation of the two isoforms of rodent NKG2D was driven by a B1 retrotransposon insertion. AB - The mouse gene for the natural killer (NK) cell-activating receptor Nkg2d produces two protein isoforms, NKG2D-S and NKG2D-L, which differ by 13 amino acids at the N-terminus and have different signalling capabilities. These two isoforms are produced through differential splicing, but their regulation has not been investigated. In this study, we show that rat Nkg2d has the same splicing pattern as that of the mouse, and we mapped transcriptional start sites in both species. We found that the splice forms arise from alternative promoters and that the NKG2D-L promoter is derived from a rodent B1 retrotransposon that inserted before mouse-rat divergence. This B1 insertion is associated with loss of a nearby splice acceptor site that subsequently allowed creation of the short NKG2D isoform found in mouse but not human. Transient reporter assays indicate that the B1 element is a strong promoter with no inherent lymphoid tissue-specificity. We have also identified different binding sites for the ETS family member GABP within both the mouse and rat B1 elements that are necessary for high-promoter activity and for full Nkg2d-L expression. These findings demonstrate that a retroelement insertion has led to gene-regulatory change and functional diversification of rodent NKG2D. PMID- 19304756 TI - Dazap2 modulates transcription driven by the Wnt effector TCF-4. AB - A major outcome of the canonical Wnt/beta-catenin-signalling pathway is the transcriptional activation of a specific set of target genes. A typical feature of the transcriptional response induced by Wnt signalling is the involvement of Tcf/Lef factors that function in the nucleus as the principal mediators of signalling. Vertebrate Tcf/Lef proteins perform two well-characterized functions: in association with beta-catenin they activate gene expression, and in the absence of Wnt ligands they bind TLE/Groucho proteins to act as transcriptional repressors. Although the general characteristics of Tcf/Lef factors are well understood, the mechanisms that control their specific roles in various cellular backgrounds are much less defined. In this report we reveal that the evolutionary conserved Dazap2 protein functions as a TCF-4 interacting partner. We demonstrate that a short region proximal to the TCF-4 HMG box mediates the interaction and that all Tcf/Lef family members associate with Dazap2. Interestingly, knockdown of Dazap2 not only reduced the activity of Wnt signalling as measured by Tcf/beta catenin reporters but additionally altered the expression of Wnt-signalling target genes. Finally, chromatin immunoprecipitation studies indicate that Dazap2 modulates the affinity of TCF-4 for its DNA-recognition motif. PMID- 19304757 TI - Creation of a type IIS restriction endonuclease with a long recognition sequence. AB - Type IIS restriction endonucleases cleave DNA outside their recognition sequences, and are therefore particularly useful in the assembly of DNA from smaller fragments. A limitation of type IIS restriction endonucleases in assembly of long DNA sequences is the relative abundance of their target sites. To facilitate ligation-based assembly of extremely long pieces of DNA, we have engineered a new type IIS restriction endonuclease that combines the specificity of the homing endonuclease I-SceI with the type IIS cleavage pattern of FokI. We linked a non-cleaving mutant of I-SceI, which conveys to the chimeric enzyme its specificity for an 18-bp DNA sequence, to the catalytic domain of FokI, which cuts DNA at a defined site outside the target site. Whereas previously described chimeric endonucleases do not produce type IIS-like precise DNA overhangs suitable for ligation, our chimeric endonuclease cleaves double-stranded DNA exactly 2 and 6 nt from the target site to generate homogeneous, 5', four-base overhangs, which can be ligated with 90% fidelity. We anticipate that these enzymes will be particularly useful in manipulation of DNA fragments larger than a thousand bases, which are very likely to contain target sites for all natural type IIS restriction endonucleases. PMID- 19304758 TI - Structural and functional multiplicity of the kisspeptin/GPR54 system in goldfish (Carassius auratus). AB - To ascertain the neuroendocrine function of the kisspeptin/GPR54 system in non mammalian species, full-length cDNAs encoding for Kiss1 and Kiss2 as well as their putative cognate receptors GPR54a and GPR54b, were isolated from goldfish (Carassius auratus). The deduced protein sequences between Kiss1 and Kiss2 in goldfish share very low similarity, but their putative mature peptides (kisspeptin-10) are relatively conserved. RT-PCR analysis demonstrated that the goldfish kiss1 gene (gfkiss1) is highly expressed in the optic tectum-thalamus, intestine, kidney, and testis, while the goldfish kiss2 gene (gfkiss2) is mainly detected in the hypothalamus, telencephalon, optic tectum thalamus, adipose tissue, kidney, heart, and gonads. The two receptor genes (gfgpr54a and gfgpr54b) are highly expressed in the brain regions including telencephalon, optic tectum thalamus, and hypothalamus. Both mature goldfish kisspeptin-10 peptides (gfKiss1 10 and gfKiss2-10) are biologically active as they could functionally interact with the two goldfish receptors expressed in cultured eukaryotic cells to trigger the downstream signaling pathways with different potencies. The actions of gfKiss1-10 and gfKiss2-10 on LH secretion were further investigated in vitro and in vivo. Intraperitoneal administration of gfKiss1-10 to sexually mature female goldfish could increase the serum LH levels. However, this peptide does not significantly influence LH release from goldfish pituitary cells in primary culture, indicating that the peptide does not exert its actions at the pituitary level. On the other hand, gfKiss2-10 appears to be a much less potent peptide as it exhibits no significant in vivo bioactivity and is also inactive on the primary pituitary cells. PMID- 19304759 TI - Shedding light on the intricate puzzle of ghrelin's effects on appetite regulation. AB - Ghrelin, a hormone primarily produced by the stomach, has a wide range of metabolic and non-metabolic effects. It also stimulates food intake through activation of various hypothalamic and brain stem neurons. A series of recent studies have explored the intracellular mechanisms of the appetite-inducing effect of ghrelin in the hypothalamus, shedding light on the intricate mechanisms of appetite regulation. AMP-activated protein kinase (AMPK) is a key metabolic enzyme involved in appetite regulation. Calmodulin kinase kinase 2 (CaMKK2) has been identified as an upstream kinase of AMPK and a key mediator in the effect of ghrelin on AMPK activity. The fatty acid pathway, hypothalamic mitochondrial respiration, and uncoupling protein 2 have been outlined as downstream targets of AMPK and mediators of ghrelin's appetite stimulating effect. This short overview summarises the present data in this field. PMID- 19304760 TI - Dentoalveolar development in subjects with normal occlusion. A longitudinal study between the ages of 5 and 31 years. AB - The aim of the present study was to examine dentoalveolar development in subjects with an 'ideal' (normal) occlusion. The material comprised 436 study casts of 189 male and 247 female subjects of Swedish origin between the ages of 5 and 31 years with no history of orthodontic treatment. Tooth width, and arch length, width, and depth, as well as palatal height were measured. The data were analysed with a Student's t-test. The results verified that continuous changes of the dental arches occur from the primary until the adult period, with individual variations. This change could be interpreted as a biological migration of the dentition, resulting in anterior crowding especially in the mandible, even in subjects with congenitally missing third molars. The occlusion should be regarded as a dynamic rather than a stable interrelationship between facial structures. This natural development has to be considered in orthodontic treatment planning as well as in assessment of stability following orthodontic treatment. A continuous increase of palatal height up to adulthood seems to be an effect of a slow continuous eruption of the teeth. This finding is also of significance in explaining the infraposition of implant-supported crowns. PMID- 19304761 TI - Analysis of supra- and subgingival long-term biofilm formation on orthodontic bands. AB - Insertion of fixed orthodontic appliances induces increased biofilm formation caused by a higher number of plaque-retentive sites. The purpose of the study was to perform a quantitative analysis of supra- and subgingival long-term biofilm formation on orthodontic bands. Ten patients (five females and five males, aged 18.3+/-5.4 years) who had received therapy with fixed orthodontic appliances for 24+/-9 months were enrolled in the study. Biofilm formation on 28 orthodontic bands was analyzed quantitatively with the Rutherford backscattering detection method, a scanning electron microscopy technique. The biofilm formation for the supra- and subgingival surfaces was calculated from the grey values. Statistical analysis was performed with a mixed model with the patient as the random factor. A P-value <0.05 was considered significant. A biofilm was found on 16.1+/-9.2 per cent of supragingival surfaces and on 3.6+/-4.4 per cent of subgingival surfaces. Differences in biofilm formation in supra- and subgingival surfaces were statistically significant (P<0.05) and formed a distinct demarcation line. Despite the presence of supragingival biofilm, no mature subgingival biofilm was found on the tested orthodontic bands. PMID- 19304763 TI - Role of plasma membrane calcium ATPases in calcium clearance from olfactory sensory neurons. AB - Odorants cause Ca(2+) to rise in olfactory sensory neurons (OSNs) first within the ciliary compartment, then in the dendritic knob, and finally in the cell body. Ca(2+) not only excites but also produces negative feedback on the transduction pathway. To relieve this Ca(2+)-dependent adaptation, Ca(2+) must be cleared from the cilia and dendritic knob by mechanisms that are not well understood. This work focuses on the roles of plasma membrane calcium pumps (PMCAs) through the use of inhibitors and mice missing 1 of the 4 PMCA isoforms (PMCA2). We demonstrate a significant contribution of PMCAs in addition to contributions of the Na(+)/Ca(2+) exchanger and endoplasmic reticulum (ER) calcium pump to the rate of calcium clearance after OSN stimulation. PMCAs in neurons can shape the Ca(2+) signal. We discuss the contributions of the specific PMCA isoforms to the shape of the Ca(2+) transient that controls signaling and adaptation in OSNs. PMID- 19304764 TI - Morphological and physiological characteristics of the serotonin-immunoreactive neuron in the antennal lobe of the male oriental tobacco budworm, Helicoverpa assulta. AB - We have characterized, by intracellular recording and staining combined with immunocytochemistry, a serotonin-immunoreactive neuron in the central olfactory pathway of the male moth Helicoverpa assulta. The neuron joins the unique category of so-called SI antennal-lobe neurons, previously described in several insect species. In similarity with that originally discovered in the sphinx moth Manduca sexta, the neuron identified here has a large soma located posteriorly in the lateral cell cluster of the antennal lobe and an unbranched neurite projecting into the ipsilateral protocerebrum via the inner antennocerebral tract. After bypassing the central body, the axon crosses the midline and extends through the corresponding antennocerebral tract to the contralateral antennal lobe where it innervates the entire assembly of glomeruli including the male specific macroglomerular complex. The neuron arborizes into several fine branches in bilateral protocerebral regions anterior to the calyces of the mushroom bodies, particularly on the contralateral side. The physiology of the neuron revealed 2 distinctly different spiking amplitudes, 1 small showing a relatively high spontaneous activity and 1 large showing low activity. The small-amplitude spikes displayed increased frequency when pheromones and plant odors were blown over the antenna. The large-amplitude spikes, which had an unusually long duration, showed no observable responses. PMID- 19304765 TI - Collaboration on children's health has some way to go, experts warn. PMID- 19304766 TI - Chronic constipation in adults. PMID- 19304767 TI - Scientists call for more funding for research into Alzheimer's disease. PMID- 19304768 TI - Reducing harm from alcohol. PMID- 19304769 TI - Spain could be 25,000 specialists short by 2025. PMID- 19304770 TI - Validation of a Monte Carlo efficiency calibration procedure for a partial body counter system with a voxel model of the LLNL torso phantom. AB - Virtual models of real phantoms used with Monte Carlo methods facilitate the calibration and other studies associated with whole-body and partial-body counting systems. In this investigation, a voxel model of an LLNL torso phantom, available physically in the in vivo laboratory at KIT, was created from computed tomography scans. Series of measurements with a high-purity germanium detector and the real torso phantom, loaded with different radioactive organs, have been carried out. Computer simulations of these measurement setups were performed with the aid of MCNPX, using a coarsened voxel phantom and a validated model of the germanium detector. The results of simulations were compared with data from the measurements and an agreement within the uncertainties was found. The voxel model could therefore be validated. The results of the simulations were then used to quantify the activity of (241)Am impurities detected in the liver loaded with (239)Pu. PMID- 19304771 TI - Evidence for allosteric interactions of antagonist binding to the smoothened receptor. AB - The Smoothened receptor (Smo) mediates hedgehog (Hh) signaling critical for development, cell growth, and migration, as well as stem cell maintenance. Aberrant Hh signaling pathway activation has been implicated in a variety of cancers, and small-molecule antagonists of Smo have entered human clinical trials for the treatment of cancer. Here, we report the biochemical characterization of allosteric interactions of agonists and antagonists for Smo. Binding of two radioligands, [(3)H]3-chloro-N-[trans-4-(methylamino)cyclohexyl]-N-{[3-(4 pyridinyl)-phenyl]methyl}-1-benzothiophene-2-carboxamide (SAG-1.3) (agonist) and [(3)H]cyclopamine (antagonist), was characterized using human Smo expressed in human embryonic kidney 293F membranes. We observed full displacement of [(3)H]cyclopamine by all Smo agonist and antagonist ligands examined. N-[(1E) (3,5-Dimethyl-1-phenyl-1H-pyrazol-4-yl)methylidene]-4-(phenylmethyl)-1 piperazinamine (SANT-1), an antagonist, did not fully inhibit the binding of [(3)H]SAG-1.3. In a functional cell-based beta-lactamase reporter gene assay, SANT-1 and N-[3-(1H-benzimidazol-2-yl)-4-chlorophenyl]-3,4,5-tris(ethyloxy) benzamide (SANT-2) fully inhibited 3-chloro-4,7-difluoro-N-[trans-4 (methylamino)cyclohexyl]-N-{[3-(4-pyridinyl)phenyl]methyl}-1-benzothiophene-2 carboxamide (SAG-1.5)-induced Hh pathway activation. Detailed "Schild-type" radioligand binding analysis with [(3)H]SAG-1.3 revealed that two structurally distinct Smoothened receptor antagonists, SANT-1 and SANT-2, bound in a manner consistent with that of allosteric modulation. Our mechanism of action characterization of radioligand binding to Smo combined with functional data provides a better understanding of small-molecule interactions with Smo and their influence on the Hh pathway. PMID- 19304772 TI - Prolonged survival with left atrial spindle cell sarcoma. AB - Primary spindle cell sarcoma of the left atrium is a rare tumour. Optimal treatment is to obtain complete surgical clearance of the tumour. The anatomic location of the tumour, infiltration into vital structures and difficult access provides a surgical challenge for resection of the lesion and reconstruction of the defect. The prognosis of patients with a primary cardiac sarcoma is very poor because of their resistance to treatment with chemotherapy and radiotherapy. Metastases and local recurrences are common despite optimal multimodality treatment. This report describes a 48-year-old gentleman who underwent multiple surgeries to achieve an 11-year survival since the diagnosis. The operative techniques have been described. PMID- 19304773 TI - Apolipoprotein E alleles and bicuspid aortic valve stenosis in monozygotic twins. AB - Aortic valve disease is a common valvular heart disease but the underlying pathology that leads to the severe dysfunction of the aortic valve is unclear. There is increasing interest in the role of hypercholesterolaemia in the causation and progression of aortic valve disease. Apolipoprotein E is an essential component of cholesterol and previous studies have reported conflicting results on the association between various apolipoprotein E alleles and aortic valve disease. We report two interesting cases of severe dysfunction of bicuspid aortic valves in twin brothers who presented to us at the same time. PMID- 19304774 TI - eComment: Plasmapheresis for near-fatal heparin-induced thrombocytopenia. PMID- 19304775 TI - eComment: Diagnostic and surgical considerations in Swyer-James-MacLeod syndrome. PMID- 19304776 TI - eComment: Swyer-James-MacLeod syndrome: addendum to the clinical appearance. PMID- 19304777 TI - eComment: Endovascular repair of iliac aneurysms. PMID- 19304779 TI - Brief report: Adherence to fluid recommendations in children receiving treatment for retentive encopresis. AB - OBJECTIVE: Limited data are available regarding whether children being treated for retentive encopresis are adherent to recommendations to increase their daily fluid intake. The purpose of this study was to examine fluid adherence in children who received treatment for retentive encopresis. METHODS: A retrospective chart review was performed using diet diary data for 26 children (ages 3-12) who completed a group behavioral intervention for retentive encopresis. RESULTS: Mean daily intake of clear fluid increased significantly during treatment and children relied primarily on water and juice to make this dietary change. However, adherence rates to clear fluid goals were <50%. CONCLUSIONS: Children's increased clear fluid intake did not equate to high fluid adherence. Children's high juice consumption is concerning as it could place them at risk for other negative health consequences. Future research should examine whether enhanced fluid education and use of behavior change strategies yield higher fluid adherence. PMID- 19304780 TI - Genome-wide scan identifies CDH13 as a novel susceptibility locus contributing to blood pressure determination in two European populations. AB - Hypertension is a complex disease that affects a large proportion of adult population. Although approximately half of the inter-individual variance in blood pressure (BP) level is heritable, identification of genes responsible for its regulation has remained challenging. Genome-wide association study (GWAS) is a novel approach to search for genetic variants contributing to complex diseases. We conducted GWAS for three BP traits [systolic and diastolic blood pressure (SBP and DBP); hypertension (HYP)] in the Kooperative Gesundheitsforschung in der Region Augsburg (KORA) S3 cohort (n = 1644) recruited from general population in Southern Germany. GWAS with 395,912 single nucleotide polymorphisms (SNPs) identified an association between BP traits and a common variant rs11646213 (T/A) upstream of the CDH13 gene at 16q23.3. The initial associations with HYP and DBP were confirmed in two other European population-based cohorts: KORA S4 (Germans) and HYPEST (Estonians). The associations between rs11646213 and three BP traits were replicated in combined analyses (dominant model: DBP, P = 5.55 x 10(-5), effect -1.40 mmHg; SBP, P = 0.007, effect -1.56 mmHg; HYP, P = 5.30 x 10(-8), OR = 0.67). Carriers of the minor allele A had a decreased risk of hypertension. A non-significant trend for association was also detected with severe family based hypertension in the BRIGHT sample (British). The novel susceptibility locus, CDH13, encodes for an adhesion glycoprotein T-cadherin, a regulator of vascular wall remodeling and angiogenesis. Its function is compatible with the BP biology and may improve the understanding of the pathogenesis of hypertension. PMID- 19304781 TI - Loss of the imprinted snoRNA mbii-52 leads to increased 5htr2c pre-RNA editing and altered 5HT2CR-mediated behaviour. AB - The Prader-Willi syndrome (PWS) genetic interval contains several brain-expressed small nucleolar (sno)RNA species that are subject to genomic imprinting. In vitro studies have shown that one of these snoRNA molecules, h/mbii-52, negatively regulates editing and alternative splicing of the serotonin 2C receptor (5htr2c) pre-RNA. However, the functional consequences of loss of h/mbii-52 and subsequent increased post-transcriptional modification of 5htr2c are unknown. 5HT2CRs are important in controlling aspects of cognition and the cessation of feeding, and disruption of their function may underlie some of the psychiatric and feeding abnormalities seen in PWS. In a mouse model for PWS lacking expression of mbii-52 (PWS-IC+/-), we show an increase in editing, but not alternative splicing, of the 5htr2c pre-RNA. This change in post-transcriptional modification is associated with alterations in a number of 5HT2CR-related behaviours, including impulsive responding, locomotor activity and reactivity to palatable foodstuffs. In a non 5HT2CR-related behaviour, marble burying, loss of mbii-52 was without effect. The specificity of the behavioural effects to changes in 5HT2CR function was further confirmed using drug challenges. These data illustrate, for the first time, the physiological consequences of altered RNA editing of 5htr2c linked to mbii-52 loss that may underlie specific aspects of the complex PWS phenotype and point to an important functional role for this imprinted snoRNA. PMID- 19304782 TI - Admixture mapping of quantitative trait loci for blood lipids in African Americans. AB - Blood lipid levels, including low-density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and triglycerides (TG), are highly heritable traits and major risk factors for atherosclerotic cardiovascular disease (CVD). Using individual ancestry estimates at marker locations across the genome, we present a novel quantitative admixture mapping analysis of all three lipid traits in a large sample of African-Americans from the Family Blood Pressure Program. Regression analysis was performed with both total and marker location-specific European ancestry as explanatory variables, along with demographic covariates. Robust permutation analysis was used to assess statistical significance. Overall European ancestry was significantly correlated with HDL-C (negatively) and TG (positively), but not with LDL-C. We found strong evidence for a novel locus underlying HDL-C on chromosome 8q, which correlated negatively with European ancestry (P = .0014); the same location also showed positive correlation of European ancestry with TG levels. A region on chromosome 14q also showed significant negative correlation between HDL-C levels and European ancestry. On chromosome 15q, a suggestive negative correlation of European ancestry with TG and positive correlation with HDL-C was observed. Results with LDL-C were less significant overall. We also found significant evidence for genome-wide ancestry effects underlying the joint distribution of HDL-C and TG, not fully explained by the locus on chromosome 8. Our results are consistent with a genetic contribution to and may explain the healthier HDL-C and TG profiles found in Blacks versus Whites. The identified regions provide locations for follow-up studies of genetic variants underlying lipid variation in African-Americans and possibly other populations. PMID- 19304783 TI - Astrocytic protection of spinal motor neurons but not cortical neurons against loss of Als2/alsin function. AB - Three neurodegenerative diseases affecting upper and/or lower motor neurons have been associated with loss of ALS2/Alsin function: juvenile amyotrophic lateral sclerosis, primary lateral sclerosis and infantile-onset ascending hereditary spastic paralysis. The distinct neuronal vulnerability and the role of glia in these diseases remains, however, unclear. We here demonstrate that alsin-depleted spinal motor neurons can be rescued from defective survival and axon growth by co cultured astrocytes. The astrocytic rescue is mediated by a soluble protective factor rather than by cellular contact. Cortical neurons are intrinsically as vulnerable to alsin depletion as spinal motor neurons but cannot be rescued by co cultured astrocytes. To our knowledge, these data provide the first example of non-cell-autonomous glial effects in a recessive form of motor neuron disease and a potential rationale for the higher vulnerability of upper versus lower motor neurons in ALS2/Alsin-linked disorders. PMID- 19304785 TI - Bypassing primary care facilities for childbirth: a population-based study in rural Tanzania. AB - In an effort to reduce maternal mortality, developing countries have been investing in village-level primary care facilities to bring skilled delivery services closer to women. We explored the extent to which women in rural western Tanzania bypass their nearest primary care facilities to deliver at more distant health facilities, using a population-representative survey of households (N = 1204). Using a standardized instrument, we asked women who had a delivery within 5 years about the place of their most recent delivery. Information on all functioning health facilities in the area were obtained from the district health office. Women who delivered in a health facility that was not the nearest available facility were considered bypassers. Forty-four per cent (186/423) of women who delivered in a health facility bypassed their nearest facility. In adjusted analysis, women who bypassed were more likely than women who did not bypass to be 35 or older (OR 2.5, P 3)-beta-D-Galp-(1[alpha-D-Glcp-(1-->6)]-->4)-beta-D ManpNAc-(1-->] repeating units that is linked by an adaptor of -[GroA-2-->OPO2- >4-beta-D-ManpNAc-(1-->4)]-->3)-alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->3)-alpha-L Rhap-(1-->3)-beta-D-Galp-(1--> to specific tyrosine residues of the S-layer protein. For elucidation of the mechanism governing S-layer glycan biosynthesis, a gene knockout system using bacterial mobile group II intron-mediated gene disruption was developed. The system is further based on the sgsE S-layer gene promoter of Geobacillus stearothermophilus NRS 2004/3a and on the Geobacillus Bacillus-Escherichia coli shuttle vector pNW33N. As a target gene, wsfP, encoding a putative UDP-Gal:phosphoryl-polyprenol Gal-1-phosphate transferase, representing the predicted initiation enzyme of S-layer glycan biosynthesis, was disrupted. S-layer protein glycosylation was completely abolished in the insertional P. alvei CCM 2051T wsfP mutant, according to sodium dodecyl sulfate polyacrylamide gel electrophoresis evidence and carbohydrate analysis. Glycosylation was fully restored by plasmid-based expression of wsfP in the glycan-deficient P. alvei mutant, confirming that WsfP initiates S-layer protein glycosylation. This is the first report on the successful genetic manipulation of bacterial S-layer protein glycosylation in vivo, including transformation of and heterologous gene expression and gene disruption in the model organism P. alvei CCM 2051T. PMID- 19304821 TI - Empirical leucine-to-carbon conversion factors for estimating heterotrophic bacterial production: seasonality and predictability in a temperate coastal ecosystem. AB - Leucine-to-carbon conversion factors (CFs) are needed for converting substrate incorporation into biomass production of heterotrophic bacteria. During 2006 we performed 20 dilution experiments for determining the spatiotemporal variability of empirical CFs in temperate Atlantic coastal waters. Values (0.49 to 1.92 kg C mol Leu(-1)) showed maxima in autumn to early winter and minima in summer. Spatially averaged CFs were significantly negatively correlated with in situ leucine incorporation rates (r = -0.91) and positively correlated with phosphate concentrations (r = 0.76). These relationships, together with a strong positive covariation between cell-specific leucine incorporation rates and carbon contents (r = 0.85), were interpreted as a strategy to maximize survival through protein synthesis and low growth rates under nutrient limitation (low CFs) until favorable conditions stimulate cell division relative to protein synthesis (high CFs). A multiple regression with in situ leucine incorporation rates and cellular carbon contents explained 96% of CF variance in our ecosystem, suggesting their potential prediction from more easily measurable routine variables. The use of the theoretical CF of 1.55 kg C mol Leu(-1) would have resulted in a serious overestimation (73%) of annual bacterial production rates. Our results emphasize the need for considering the temporal scale in CFs for bacterial production studies. PMID- 19304820 TI - Effect of lake trophic status and rooted macrophytes on community composition and abundance of ammonia-oxidizing prokaryotes in freshwater sediments. AB - Communities of ammonia-oxidizing archaea (AOA) and bacteria (AOB) in freshwater sediments and those in association with the root system of the macrophyte species Littorella uniflora, Juncus bulbosus, and Myriophyllum alterniflorum were compared for seven oligotrophic to mesotrophic softwater lakes and acidic heathland pools. Archaeal and bacterial ammonia monooxygenase alpha-subunit (amoA) gene diversity increased from oligotrophic to mesotrophic sites; the number of detected operational taxonomic units was positively correlated to ammonia availability and pH and negatively correlated to sediment C/N ratios. AOA communities could be grouped according to lake trophic status and pH; plant species-specific communities were not detected, and no grouping was apparent for AOB communities. Relative abundance, determined by quantitative PCR targeting amoA, was always low for AOB (<0.05% of all prokaryotes) and slightly higher for AOA in unvegetated sediment and AOA in association with M. alterniflorum (0.01 to 2%), while AOA accounted for up to 5% in the rhizospheres of L. uniflora and J. bulbosus. These results indicate that (i) AOA are at least as numerous as AOB in freshwater sediments, (ii) aquatic macrophytes with substantial release of oxygen and organic carbon into their rhizospheres, like L. uniflora and J. bulbosus, increase AOA abundance; and (iii) AOA community composition is generally determined by lake trophy, not by plant species-specific interactions. PMID- 19304822 TI - Influence of stress on single-cell lag time and growth probability for Listeria monocytogenes in half Fraser broth. AB - The impacts of 12 common food industry stresses on the single-cell growth probability and single-cell lag time distribution of Listeria monocytogenes were determined in half Fraser broth, the primary enrichment broth of the International Organization for Standardization detection method. First, it was determined that the ability of a cell to multiply in half Fraser broth is conditioned by its history (the probability for a cell to multiply can be decreased to 0.05), meaning that, depending on the stress in question, the risk of false-negative samples can be very high. Second, it was established that when cells are injured, the single-cell lag times increase in mean and in variability and that this increase represents a true risk of not reaching the detection threshold of the method in the enrichment broth. No relationship was observed between the impact on single-cell lag times and that on growth probabilities. These results emphasize the importance of taking into account the physiological state of the cells when evaluating the performance of methods to detect pathogens in food. PMID- 19304823 TI - Exopolyphosphatases PPX1 and PPX2 from Corynebacterium glutamicum. AB - Corynebacterium glutamicum accumulates up to 300 mM of inorganic polyphosphate (PolyP) in the cytosol or in granules. The gene products of cg0488 (ppx1) and cg1115 (ppx2) were shown to be active as exopolyphosphatases (PPX), as overexpression of either gene resulted in higher exopolyphosphatase activities in crude extracts and deletion of either gene with lower activities than those of the wild-type strain. PPX1 and PPX2 from C. glutamicum share only 25% identical amino acids and belong to different protein groups, which are distinct from enterobacterial, archaeal, and yeast exopolyphosphatases. In comparison to that in the wild type, more intracellular PolyP accumulated in the Deltappx1 and Deltappx2 deletion mutations but less when either ppx1 or ppx2 was overexpressed. When C. glutamicum was shifted from phosphate-rich to phosphate-limiting conditions, a growth advantage of the deletion mutants and a growth disadvantage of the overexpression strains compared to the wild type were observed. Growth experiments, exopolyphosphatase activities, and intracellular PolyP concentrations revealed PPX2 as being a major exopolyphosphatase from C. glutamicum. PPX2(His) was purified to homogeneity and shown to be active as a monomer. The enzyme required Mg2+ or Mn2+ cations but was inhibited by millimolar concentrations of Mg2+, Mn2+, and Ca2+. PPX2 from C. glutamicum was active with short-chain polyphosphates, even accepting pyrophosphate, and was inhibited by nucleoside triphosphates. PMID- 19304824 TI - Failure to detect Helicobacter pylori DNA in drinking and environmental water in Dhaka, Bangladesh, using highly sensitive real-time PCR assays. AB - The main transmission pathway of Helicobacter pylori has not been determined, but several reports have described detection of H. pylori DNA in drinking and environmental water, suggesting that H. pylori may be waterborne. To address this possibility, we developed, tested, and optimized two complementary H. pylori specific real-time PCR assays for quantification of H. pylori DNA in water. The minimum detection level of the assays including collection procedures and DNA extraction was shown to be approximately 250 H. pylori genomes per water sample. Using our assays, we then analyzed samples of drinking and environmental water (n = 75) and natural water biofilms (n = 21) from a high-endemicity area in Bangladesh. We could not identify H. pylori DNA in any of the samples, even though other pathogenic bacteria have been found previously in the same water samples by using the same methodology. A series of control experiments were performed to ensure that the negative results were not falsely caused by PCR inhibition, nonspecific assays, degradation of template DNA, or low detection sensitivity. Our results suggest that it is unlikely that the predominant transmission route of H. pylori in this area is waterborne. PMID- 19304825 TI - Light-dependent transcriptional regulation of genes of biogeochemical interest in the diploid and haploid life cycle stages of Emiliania huxleyi. AB - The expression of genes of biogeochemical interest in calcifying and noncalcifying life stages of the coccolithophore Emiliania huxleyi was investigated. Transcripts potentially involved in calcification were tested through a light-dark cycle. These transcripts were more abundant in calcifying cells and were upregulated in the light. Their application as potential candidates for in situ biogeochemical proxies is also suggested. PMID- 19304826 TI - Identification of natural target proteins indicates functions of a serralysin type metalloprotease, PrtA, in anti-immune mechanisms. AB - Serralysins are generally thought to function as pathogenicity factors of bacteria, but so far no hard evidence of this (e.g., specific substrate proteins that are sensitive to the cleavage by these proteases) has been found. We have looked for substrate proteins to a serralysin-type proteinase, PrtA, in a natural host-pathogen molecular interaction system involving Manduca sexta and Photorhabdus luminescens. The exposure in vitro of hemolymph to PrtA digestion resulted in selective cleavage of 16 proteins, provisionally termed PAT (PrtA target) proteins. We could obtain sequence information for nine of these PrtA sensitive proteins, and by searching databases, we could identify six of them. Each has immune-related function involving every aspect of the immune defense: beta-1,3 glucan recognition protein 2 (immune recognition), hemocyte aggregation inhibitor protein (HAIP), serine proteinase homolog 3, six serpin-1 variants, including serpin-1I (immune signaling and regulation), and scolexins A and B (coagulation cascade effector function). The functions of the identified PrtA substrate proteins shed new light on a possible participation of a serralysin in the virulence mechanism of a pathogen. Provided these proteins are targets of PrtA in vivo, this might represent, among others, a complex suppressive role on the innate immune response via interference with both the recognition and the elimination of the pathogen during the first, infective stage of the host pathogen interaction. Our results also raise the possibility that the natural substrate proteins of serralysins of vertebrate pathogens might be found among the components of the innate immune system. PMID- 19304827 TI - Differential responses of nitrate reducer community size, structure, and activity to tillage systems. AB - The main objective of this study was to determine how the size, structure, and activity of the nitrate reducer community were affected by adoption of a conservative tillage system as an alternative to conventional tillage. The experimental field, established in Madagascar in 1991, consists of plots subjected to conventional tillage or direct-seeding mulch-based cropping systems (DM), both amended with three different fertilization regimes. Comparisons of size, structure, and activity of the nitrate reducer community in samples collected from the top layer in 2005 and 2006 revealed that all characteristics of this functional community were affected by the tillage system, with increased nitrate reduction activity and numbers of nitrate reducers under DM. Nitrate reduction activity was also stimulated by combined organic and mineral fertilization but not by organic fertilization alone. In contrast, both negative and positive effects of combined organic and mineral fertilization on the size of the nitrate reducer community were observed. The size of the nitrate reducer community was a significant predictor of the nitrate reduction rates except in one treatment, which highlighted the inherent complexities in understanding the relationships the between size, diversity, and structure of functional microbial communities along environmental gradients. PMID- 19304828 TI - Development of cpn60-based real-time quantitative PCR assays for the detection of 14 Campylobacter species and application to screening of canine fecal samples. AB - Campylobacter species are important organisms in both human and animal health. The identification of Campylobacter currently requires the growth of organisms from complex samples and biochemical identification. In many cases, the condition of the sample being tested and/or the fastidious nature of many Campylobacter species has limited the detection of campylobacters in a laboratory setting. To address this, we have designed a set of real-time quantitative PCR (qPCR) assays to detect and quantify 14 Campylobacter species, C. coli, C. concisus, C. curvus, C. fetus, C. gracilis, C. helveticus, C. hyointestinalis, C. jejuni, C. lari, C. mucosalis, C. rectus, C. showae, C. sputorum, and C. upsaliensis, directly from DNA extracted from feces. By use of a region of the cpn60 (also known as hsp60 or groEL) gene, which encodes the universally conserved 60-kDa chaperonin, species specific assays were designed and validated. These assays were then employed to determine the prevalence of Campylobacter species in fecal samples from dogs. Fecal samples were found to contain detectable and quantifiable levels of C. fetus, C. gracilis, C. helveticus, C. jejuni, C. showae, and C. upsaliensis, with the majority of samples containing multiple Campylobacter species. This study represents the first report of C. fetus, C. gracilis, C. mucosalis, and C. showae detection in dogs and implicates dogs as a reservoir for these species. The qPCR assays described offer investigators a new tool to study many Campylobacter species in a culture-independent manner. PMID- 19304829 TI - Diversity of bacteria in the marine sponge Aplysina fulva in Brazilian coastal waters. AB - Microorganisms can account for up to 60% of the fresh weight of marine sponges. Marine sponges have been hypothesized to serve as accumulation spots of particular microbial communities, but it is unknown to what extent these communities are directed by the organism or the site or occur randomly. To address this question, we assessed the composition of specific bacterial communities associated with Aplysina fulva, one of the prevalent sponge species inhabiting Brazilian waters. Specimens of A. fulva and surrounding seawater were collected in triplicate in shallow water at two sites, Caboclo Island and Tartaruga beach, Buzios, Brazil. Total community DNA was extracted from the samples using "direct" and "indirect" approaches. 16S rRNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses of the total bacterial community and of specific bacterial groups--Pseudomonas and Actinobacteria--revealed that the structure of these assemblages in A. fulva differed drastically from that observed in seawater. The DNA extraction methodology and sampling site were determinative for the composition of actinobacterial communities in A. fulva. However, no such effects could be gleaned from total bacterial and Pseudomonas PCR-DGGE profiles. Bacterial 16S rRNA gene clone libraries constructed from directly and indirectly extracted DNA did not differ significantly with respect to diversity and composition. Altogether, the libraries encompassed 15 bacterial phyla and the candidate division TM7. Clone sequences affiliated with the Cyanobacteria, Chloroflexi, Gamma- and Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were, in this order, most abundant. The bacterial communities associated with the A. fulva specimens were distinct and differed from those described in studies of sponge-associated microbiota performed with other sponge species. PMID- 19304830 TI - Genetic diversity and fitness of Fusarium graminearum populations from rice in Korea. AB - Fusarium graminearum is an important fungal pathogen of cereal crops and produces mycotoxins, such as the trichothecenes nivalenol and deoxynivalenol. This species may be subdivided into a series of genetic lineages or phylogenetic species. We identified strains of F. graminearum from the Republic of Korea to lineage, tested their ability to produce nivalenol and deoxynivalenol, and determined the genetic composition and structure of the populations from which they were recovered. Based on amplified fragment length polymorphism (AFLP), PCR genotyping, and chemical analyses of trichothecenes, all 249 isolates from southern provinces belonged to lineage 6, with 241 having the nivalenol genotype and 8 having the deoxynivalenol genotype. In the eastern Korea province, we recovered 84 lineage 6 isolates with the nivalenol genotype and 23 lineage 7 isolates with the deoxynivalenol genotype. Among 333 lineage 6 isolates, 36% of the AFLP bands were polymorphic, and there were 270 multilocus haplotypes. Genetic identity among populations was high (>0.972), and genotype diversity was low (30 to 58%). To test the adaptation of lineage 6 to rice, conidial mixtures of strains from lineages 3, 6, and 7 were inoculated onto rice plants and then recovered from the rice grains produced. Strains representing lineages 6 and 7 were recovered from inoculated spikelets at similar frequencies that were much higher than those for the strain representing lineage 3. Abundant perithecia were produced on rice straw, and 247 single-ascospore isolates were recovered from 247 perithecia. Perithecia representing lineage 6 (87%) were the most common, followed by those representing lineage 7 (13%), with perithecia representing lineage 3 not detected. These results suggest that F. graminearum lineage 6 may have a host preference for rice and that it may be more fit in a rice agroecosystem than are the other lineages present in Korea. PMID- 19304831 TI - Cloning and in situ expression studies of the Hydrogenobaculum arsenite oxidase genes. AB - Novel arsenite [As(III)] oxidase structural genes (aoxAB) were cloned from Hydrogenobaculum bacteria isolated from an acidic geothermal spring. Reverse transcriptase PCR demonstrated expression throughout the outflow channel, and the aoxB cDNA clones exhibited distribution patterns relative to the physicochemical gradients in the spring. Microelectrode analyses provided evidence of quantitative As(III) transformation within the microbial mat. PMID- 19304832 TI - Application of a newly identified and characterized 18-o-acyltransferase in chemoenzymatic synthesis of selected natural and nonnatural bioactive derivatives of phoslactomycins. AB - Phoslactomycins (PLMs) and related leustroducsins (LSNs) have been isolated from a variety of bacteria based on antifungal, anticancer, and other biological assays. Streptomyces sp. strain HK 803 produces five PLM analogs (PLM A and PLMs C to F) in which the C-18 hydroxyl substituent is esterified with a range of branched, short-alkyl-chain carboxylic acids. The proposed pathway intermediate, PLM G, in which the hydroxyl residue is not esterified has not been observed at any significant level in fermentation, and the only route to this potentially useful intermediate has been an enzymatic deacylation of other PLMs and LSNs. We report that deletion of plmS(3) from the PLM biosynthetic cluster gives rise to a mutant which accumulates the PLM G intermediate. The 921-bp plmS(3) open reading frame was cloned and expressed as an N-terminally polyhistidine-tagged protein in Escherichia coli and shown to be an 18-O acyltransferase, catalyzing conversion of PLM G to PLM A, PLM C, and PLM E using isobutyryl coenzyme A (CoA), 3 methylbutyryl-CoA, and cyclohexylcarbonyl-CoA, respectively. The efficiency of this process (k(cat) of 28 +/- 3 min(-1) and K(m) of 88 +/- 16 microM) represents a one-step chemoenzymatic alternative to a multistep synthetic process for selective chemical esterification of the C-18 hydroxy residue of PLM G. PlmS(3) was shown to catalyze esterification of PLM G with CoA and N-acetylcysteamine thioesters of various saturated, unsaturated, and aromatic carboxylic acids and thus also to provide an efficient chemoenzymatic route to new PLM analogs. PMID- 19304833 TI - Negative correlation between individual-insect-level virulence and colony-level virulence of Paenibacillus larvae, the etiological agent of American foulbrood of honeybees. AB - Paenibacillus larvae is the etiological agent of American foulbrood (AFB) in honeybees. Recently, different genotypes of P. larvae (ERIC I to ERIC IV) were defined, and it was shown that these genotypes differ inter alia in their virulence on the larval level. On the colony level, bees mitigate AFB through the hygienic behavior of nurse bees. Therefore, we investigated how the hygienic behavior shapes P. larvae virulence on the colony level. Our results indicate that P. larvae virulence on the larval level and that on the colony level are negatively correlated. PMID- 19304834 TI - Changes in benthic denitrification, nitrate ammonification, and anammox process rates and nitrate and nitrite reductase gene abundances along an estuarine nutrient gradient (the Colne estuary, United Kingdom). AB - Estuarine sediments are the location for significant bacterial removal of anthropogenically derived inorganic nitrogen, in particular nitrate, from the aquatic environment. In this study, rates of benthic denitrification (DN), dissimilatory nitrate reduction to ammonium (DNRA), and anammox (AN) at three sites along a nitrate concentration gradient in the Colne estuary, United Kingdom, were determined, and the numbers of functional genes (narG, napA, nirS, and nrfA) and corresponding transcripts encoding enzymes mediating nitrate reduction were determined by reverse transcription-quantitative PCR. In situ rates of DN and DNRA decreased toward the estuary mouth, with the findings from slurry experiments suggesting that the potential for DNRA increased while the DN potential decreased as nitrate concentrations declined. AN was detected only at the estuary head, accounting for approximately 30% of N2 formation, with 16S rRNA genes from anammox-related bacteria also detected only at this site. Numbers of narG genes declined along the estuary, while napA gene numbers were stable, suggesting that NAP-mediated nitrate reduction remained important at low nitrate concentrations. nirS gene numbers (as indicators of DN) also decreased along the estuary, whereas nrfA (an indicator for DNRA) was detected only at the two uppermost sites. Similarly, nitrate and nitrite reductase gene transcripts were detected only at the top two sites. A regression analysis of log(n + 1) process rate data and log(n + 1) mean gene abundances showed significant relationships between DN and nirS and between DNRA and nrfA. Although these log-log relationships indicate an underlying relationship between the genetic potential for nitrate reduction and the corresponding process activity, fine-scale environmentally induced changes in rates of nitrate reduction are likely to be controlled at cellular and protein levels. PMID- 19304835 TI - Inactivation of the SauI type I restriction-modification system is not sufficient to generate Staphylococcus aureus strains capable of efficiently accepting foreign DNA. AB - Genetic manipulation of Staphylococcus aureus is limited by the availability of only a single strain, RN4220, that is capable of easily accepting foreign DNA. Inactivation of the hsdR gene of the SauI type I restriction-modification system was shown previously to be responsible for the high transformation efficiency of RN4220 (D. E. Waldron and J. A. Lindsay, J Bacteriol. 188:5578-5585, 2006). However, deletion of this gene in three different S. aureus strains was not sufficient to make them readily transformable, which would be remarkably useful for genetic studies of this pathogenic organism. These results indicate that another unknown factor(s) is required for the transformable phenotype in S. aureus. PMID- 19304836 TI - Stereospecific biotransformation of dihydrodaidzein into (3S)-equol by the human intestinal bacterium Eggerthella strain Julong 732. AB - Stereochemical course of isoflavanone dihydrodaidzein (DHD) reduction into the isoflavan (3S)-equol via tetrahydrodaidzein (THD) by the human intestinal anaerobic bacterium Eggerthella strain Julong 732 was studied. THD was synthesized by catalytic hydrogenation, and each stereoisomer was separated by chiral high-performance liquid chromatography. Circular dichroism spectroscopy was used to elucidate the absolute configurations of four synthetic THD stereoisomers. Rapid racemization of DHD catalyzed by Julong 732 prevented the substrate stereospecificity in the conversion of DHD into THD from being confirmed. The absolute configuration of THD, prepared by reduction of DHD in the cell-free incubation, was assigned as (3R,4S) via comparison of the retention time to that of the authentic THD by chiral chromatography. Dehydroequol (DE) was unable to produce the (3S)-equol both in the cell-free reaction and in the bacterial transformation, negating the possible intermediacy of DE. Finally, the intermediate (3R,4S)-THD was reduced into (3S)-equol by the whole cell, indicating the inversion of stereochemistry at C-3 during the reduction. A possible mechanism accounting for the racemization of DHD and the inversion of configuration of THD during reduction into (3S)-equol is proposed. PMID- 19304837 TI - Distribution, diversity, and potential mobility of extrachromosomal elements related to the Bacillus anthracis pXO1 and pXO2 virulence plasmids. AB - The presence of a pXO1- and/or pXO2-like plasmid(s) in clinical isolates of Bacillus cereus sensu stricto and in strains of the biopesticide Bacillus thuringiensis has been reported recently, and the pXO2-like plasmid pBT9727 and another pXO2-like plasmid, pAW63, were found to be conjugative. In this study, a total of 1,000 B. cereus group isolates were analyzed for the presence of pXO1- and pXO2-like replicons and for the presence of pXO2-related conjugative modules. pXO1- and pXO2-like replicons were present in ca. 6.6% and 7.7% of random environmental samples, respectively, and ca. 1.54% of the strains were positive for pXO2-like transfer module genes. Only the strains harboring a pXO2-like replicon also contained the corresponding transfer genes. For the strains which contained a pXO1- and/or pXO2-like replicon(s), a large plasmid(s) whose size was similar to that of pXO1-like and/or pXO2-like plasmids was also observed, but none of these isolates were found to carry the Bacillus anthracis toxin or capsule virulence genes. Furthermore, 17 of 22 pXO2-like plasmids containing the transfer modules were able to self-transfer and to mobilize small plasmids. No pXO1- or pXO2-like plasmid lacking the cognate transfer modules has been found to have transfer potential. In the strains possessing the putative pXO2-like conjugative apparatus, variations in the presence of the group II introns B.th.I.1 and B.th.I.2 were observed, suggesting that there is important flexibility in the conjugation modules and their regulation. There was no consistent correlation between a pXO2-like repA dendrogram and the presence of the tra region or between a virB4 dendrogram and transfer ability. Discrepancies between pXO2-like repA and virB4 dendrograms were also observed, indicating that the evolution of pXO2 is an active process. PMID- 19304838 TI - Bile-inducible efflux transporter from Bifidobacterium longum NCC2705, conferring bile resistance. AB - Bifidobacteria are normal inhabitants of the human gut. Some strains of this genus are considered health promoting or probiotic, being included in numerous food products. In order to exert their health benefits, these bacteria must overcome biological barriers, including bile salts, to colonize and survive in specific parts of the intestinal tract. The role of multidrug resistance (MDR) transporters in bile resistance of probiotic bacteria and the effect of bile on probiotic gene expression are not fully understood. In the present study, the effect of subinhibitory concentrations of bile on the expression levels of predicted MDR genes from three different bifidobacterial strains, belonging to Bifidobacterium longum subsp. longum, Bifidobacterium breve, and Bifidobacterium animalis subsp. lactis, was tested. In this way, two putative MDR genes whose expression was induced by bile, BL0920 from B. longum and its homolog, Bbr0838, from B. breve, were identified. The expression of the BL0920 gene in Escherichia coli was shown to confer resistance to bile, likely to be mediated by active efflux from the cells. To the best of our knowledge, this represents the first identified bifidobacterial bile efflux pump whose expression is induced by bile. PMID- 19304839 TI - Cultivation of fastidious bacteria by viability staining and micromanipulation in a soil substrate membrane system. AB - Soil substrate membrane systems allow for microcultivation of fastidious soil bacteria as mixed microbial communities. We isolated established microcolonies from these membranes by using fluorescence viability staining and micromanipulation. This approach facilitated the recovery of diverse, novel isolates, including the recalcitrant bacterium Leifsonia xyli, a plant pathogen that has never been isolated outside the host. PMID- 19304840 TI - Microbial diversity of septic tank effluent and a soil biomat. AB - Microbial diversity of septic tank effluent (STE) and the biomat that is formed as a result of STE infiltration on soil were characterized by 16S rRNA gene sequence analysis. Results indicate that microbial communities are different within control soil, STE, and the biomat and that microbes found in STE are not found in the biomat. The development of a stable soil biomat appears to provide the best on-site water treatment or protection for subsequent groundwater interactions of STE. PMID- 19304841 TI - Development and application of a upp-based counterselective gene replacement system for the study of the S-layer protein SlpX of Lactobacillus acidophilus NCFM. AB - In silico genome analysis of Lactobacillus acidophilus NCFM coupled with gene expression studies have identified putative genes and regulatory networks that are potentially important to this organism's survival, persistence, and activities in the gastrointestinal tract. Correlation of key genotypes to phenotypes requires an efficient gene replacement system. In this study, use of the upp-encoded uracil phosphoribosyltransferase (UPRTase) of L. acidophilus NCFM was explored as a counterselection marker to positively select for recombinants that have resolved from chromosomal integration of pORI-based plasmids. An isogenic mutant carrying a upp gene deletion was constructed and was resistant to 5-fluorouracil (5-FU), a toxic uracil analog that is also a substrate for UPRTase. A 3.0-kb pORI-based counterselectable integration vector bearing a upp expression cassette, pTRK935, was constructed and introduced into the Deltaupp host harboring the pTRK669 helper plasmid. Extrachromosomal replication of pTRK935 complemented the mutated chromosomal upp allele and restored sensitivity to 5-FU. This host background provides a platform for a two-step plasmid integration and excision strategy that can select for plasmid-free recombinants with either the wild-type or mutated allele of the targeted gene in the presence of 5-FU. The efficacy of the system was demonstrated by in-frame deletion of the slpX gene (LBA0512) encoding a novel 51-kDa secreted protein associated with the S-layer complex of L. acidophilus. The resulting DeltaslpX mutant exhibited lower growth rates, increased sensitivity to sodium dodecyl sulfate, and greater resistance to bile. Overall, this improved gene replacement system represents a valuable tool for investigating the mechanisms underlying the probiotic functionality of L. acidophilus. PMID- 19304842 TI - Social power and approach-related neural activity. AB - It has been argued that power activates a general tendency to approach whereas powerlessness activates a tendency to inhibit. The assumption is that elevated power involves reward-rich environments, freedom and, as a consequence, triggers an approach-related motivational orientation and attention to rewards. In contrast, reduced power is associated with increased threat, punishment and social constraint and thereby activates inhibition-related motivation. Moreover, approach motivation has been found to be associated with increased relative left sided frontal brain activity, while withdrawal motivation has been associated with increased right sided activations. We measured EEG activity while subjects engaged in a task priming either high or low social power. Results show that high social power is indeed associated with greater left-frontal brain activity compared to low social power, providing the first neural evidence for the theory that high power is associated with approach-related motivation. We propose a framework accounting for differences in both approach motivation and goal directed behaviour associated with different levels of power. PMID- 19304843 TI - What motivates repayment? Neural correlates of reciprocity in the Trust Game. AB - Reciprocity of trust is important for social interaction and depends on individual differences in social value orientation (SVO). Here, we examined the neural correlates of reciprocity by manipulating two factors that influence reciprocal behavior: (1) the risk that the trustor took when trusting and (2) the benefit for the trustee when being trusted. FMRI results showed that anterior Medial Prefrontal Frontal Cortex (aMPFC) was more active when participants defected relative to when participants reciprocated, but was not sensitive to manipulations of risk and benefit or individual differences in SVO. However, activation in the temporal-parietal-junction (rTPJ), bilateral anterior insula and anterior cingulate cortex (ACC) was modulated by individual differences in SVO. In addition, these regions were differentially sensitive to manipulations of risk for the trustor when reciprocating. In contrast, the ACC and the right dorsolateral prefrontal cortex were sensitive to the benefit for the trustee when reciprocating. Together, the results of this study provide more insight in how several brain regions work together when individuals reciprocate trust, by showing how these regions are differentially sensitive to reciprocity motives and perspective-taking. PMID- 19304844 TI - Biochemical analysis of a beta-D-xylosidase and a bifunctional xylanase-ferulic acid esterase from a xylanolytic gene cluster in Prevotella ruminicola 23. AB - Prevotella ruminicola 23 is an obligate anaerobic bacterium in the phylum Bacteroidetes that contributes to hemicellulose utilization within the bovine rumen. To gain insight into the cellular machinery that this organism elaborates to degrade the hemicellulosic polymer xylan, we identified and cloned a gene predicted to encode a bifunctional xylanase-ferulic acid esterase (xyn10D-fae1A) and expressed the recombinant protein in Escherichia coli. Biochemical analysis of purified Xyn10D-Fae1A revealed that this protein possesses both endo-beta-1,4 xylanase and ferulic acid esterase activities. A putative glycoside hydrolase (GH) family 3 beta-D-glucosidase gene, with a novel PA14-like insertion sequence, was identified two genes downstream of xyn10D-fae1A. Biochemical analyses of the purified recombinant protein revealed that the putative beta-D-glucosidase has activity for pNP-beta-D-xylopyranoside, pNP-alpha-L-arabinofuranoside, and xylo oligosaccharides; thus, the gene was designated xyl3A. When incubated in combination with Xyn10D-Fae1A, Xyl3A improved the release of xylose monomers from a hemicellulosic xylan substrate, suggesting that these two enzymes function synergistically to depolymerize xylan. Directed mutagenesis studies of Xyn10D Fae1A mapped the catalytic sites for the two enzymatic functionalities to distinct regions within the polypeptide sequence. When a mutation was introduced into the putative catalytic site for the xylanase domain (E280S), the ferulic acid esterase activity increased threefold, which suggests that the two catalytic domains for Xyn10D-Fae1A are functionally coupled. Directed mutagenesis of conserved residues for Xyl3A resulted in attenuation of activity, which supports the assignment of Xyl3A as a GH family 3 beta-D-xylosidase. PMID- 19304845 TI - The biosynthetic pathway for myxol-2' fucoside (myxoxanthophyll) in the cyanobacterium Synechococcus sp. strain PCC 7002. AB - Synechococcus sp. strain PCC 7002 produces a variety of carotenoids, which comprise predominantly dicylic beta-carotene and two dicyclic xanthophylls, zeaxanthin and synechoxanthin. However, this cyanobacterium also produces a monocyclic myxoxanthophyll, which was identified as myxol-2' fucoside. Compared to the carotenoid glycosides produced by diverse microorganisms, cyanobacterial myxoxanthophyll and closely related compounds are unusual because they are glycosylated on the 2'-OH rather than on the 1'-OH position of the psi end of the molecule. In this study, the genes encoding two enzymes that modify the psi end of myxoxanthophyll in Synechococcus sp. strain PCC 7002 were identified. Mutational and biochemical studies showed that open reading frame SynPCC7002_A2032, renamed cruF, encodes a 1',2'-hydroxylase [corrected] and that open reading frame SynPCC7002_A2031, renamed cruG, encodes a 2'-O glycosyltransferase. The enzymatic activity of CruF was verified by chemical characterization of the carotenoid products synthesized when cruF was expressed in a lycopene-producing strain of Escherichia coli. Database searches showed that homologs of cruF and cruG occur in the genomes of all sequenced cyanobacterial strains that are known to produce myxol or the acylic xanthophyll oscillaxanthin. The genomes of many other bacteria that produce hydroxylated carotenoids but do not contain crtC homologs also contain cruF orthologs. Based upon observable intermediates, a complete biosynthetic pathway for myxoxanthophyll is proposed. This study expands the suite of enzymes available for metabolic engineering of carotenoid biosynthetic pathways for biotechnological applications. PMID- 19304846 TI - Redundancy of enzymes for formaldehyde detoxification in Pseudomonas putida. AB - Pseudomonas putida KT2440 exhibits redundant formaldehyde dehydrogenases and formate dehydrogenases that contribute to the detoxification of formaldehyde, a highly toxic compound. Physical and transcriptional analyses showed that the open reading frame (ORF) PP0328, encoding one of the formaldehyde dehydrogenases, is self-sufficient, whereas the other functional formaldehyde dehydrogenase gene (ORF PP3970) forms an operon with another gene of unknown function. Two formate dehydrogenase gene clusters (PP0489 to PP0492 and PP2183 to PP2186) were identified, and genes in these clusters were found to form operons. All four transcriptional promoters were mapped by primer extension and revealed the presence of noncanonical promoters expressed at basal level in the exponential growth phase and at a higher level in the stationary phase regardless of the presence of extracellular formaldehyde or formate. These promoters were characterized by a 5'-AG-CCA-C/A-CT-3' conserved region between -7 and -16. To determine the contribution of the different gene products to formaldehyde and formate mineralization, mutants with single and double mutations of formaldehyde dehydrogenases were generated, and the effect of the mutations on formaldehyde catabolism was tested by measuring (14)CO(2) evolution from (14)C-labeled formaldehyde. The results showed that both enzymes contributed to formaldehyde catabolism. A double mutant lacking these two enzymes still evolved CO(2) from formaldehyde, suggesting the presence of one or more still-unidentified formaldehyde dehydrogenases. Mutants with single and double mutations in the clusters for formate dehydrogenases were also generated, and all of them were able to metabolize [(14)C]formate to (14)CO(2), suggesting a redundancy of functions that was not limited to only the annotated genes. Single and double mutants deficient in formaldehyde dehydrogenases and formate dehydrogenases exhibited longer lag phases than did the parental strain when confronted with concentrations of formaldehyde close to the MICs. This suggests a role for the detoxification system in tolerance to sublethal concentrations of formaldehyde. PMID- 19304847 TI - Transsexuality in the rhizosphere: quorum sensing reversibly converts Agrobacterium tumefaciens from phenotypically female to male. AB - Conjugative plasmids generally encode proteins that block the conjugative entry of identical or similar plasmids into the host cell, a phenomenon known as entry exclusion. Here, we demonstrate that two Ti plasmids of Agrobacterium tumefaciens encode robust entry exclusion functions. Two proteins, TrbJ and TrbK, can each mediate entry exclusion and act synergistically. The trbJ and trbK genes are included within the trb operon, which is tightly regulated by the quorum-sensing regulator TraR and the cognate acylhomoserine lactone. In the absence of quorum sensing signals, these proteins are not significantly expressed, and cells lacking TrbJ and TrbK are efficient Ti plasmid recipients. In the presence of these signals, these strains block the entry of Ti plasmids and instead become efficient conjugal donors. PMID- 19304848 TI - Identification of a polymyxin synthetase gene cluster of Paenibacillus polymyxa and heterologous expression of the gene in Bacillus subtilis. AB - Polymyxin, a long-known peptide antibiotic, has recently been reintroduced in clinical practice because it is sometimes the only available antibiotic for the treatment of multidrug-resistant gram-negative pathogenic bacteria. Lack of information on the biosynthetic genes of polymyxin, however, has limited the study of structure-function relationships and the development of improved polymyxins. During whole genome sequencing of Paenibacillus polymyxa E681, a plant growth-promoting rhizobacterium, we identified a gene cluster encoding polymyxin synthetase. Here, we report the complete sequence of the gene cluster and its function in polymyxin biosynthesis. The gene cluster spanning the 40.6-kb region consists of five open reading frames, designated pmxA, pmxB, pmxC, pmxD, and pmxE. The pmxC and pmxD genes are similar to genes that encode transport proteins, while pmxA, pmxB, and pmxE encode polymyxin synthetases. The insertional disruption of pmxE led to a loss of the ability to produce polymyxin. Introduction of the pmx gene cluster into the amyE locus of the Bacillus subtilis chromosome resulted in the production of polymyxin in the presence of extracellularly added L-2,4-diaminobutyric acid. Taken together, our findings demonstrate that the pmx gene cluster is responsible for polymyxin biosynthesis. PMID- 19304849 TI - Surface location of individual residues of SlpA provides insight into the Lactobacillus brevis S-layer. AB - Bacterial surface layer (S-layer) proteins are excellent candidates for in vivo and in vitro nanobiotechnological applications because of their ability to self assemble into two-dimensional lattices that form the outermost layer of many Eubacteria and most Archaea species. Despite this potential, knowledge about their molecular architecture is limited. In this study, we investigated SlpA, the S-layer protein of the potentially probiotic bacterium Lactobacillus brevis ATCC 8287 by cysteine-scanning mutagenesis and chemical modification. We generated a series of 46 mutant proteins by replacing single amino acids with cysteine, which is not present in the wild-type protein. Most of the replaced amino acids were located in the self-assembly domain (residues 179 to 435) that likely faces the outer surface of the lattice. As revealed by electron microscopy, all the mutant proteins were able to form self-assembly products identical to that of the wild type, proving that this replacement does not dramatically alter the protein conformation. The surface accessibility of the sulfhydryl groups introduced was studied with two maleimide-containing marker molecules, TMM(PEG)(12) (molecular weight [MW], 2,360) and AlexaFluor488-maleimide (MW = 720), using both monomeric proteins in solution and proteins allowed to self-assemble on cell wall fragments. Using the acquired data and available domain information, we assigned the mutated residues into four groups according to their location in the protein monomer and lattice structure: outer surface of the lattice (9 residues), inner surface of the lattice (9), protein interior (12), and protein-protein interface/pore regions (16). This information is essential, e.g., in the development of therapeutic and other health-related applications of Lactobacillus S-layers. PMID- 19304850 TI - Characterization of a novel thermostable carboxylesterase from Geobacillus kaustophilus HTA426 shows the existence of a new carboxylesterase family. AB - The gene GK3045 (741 bp) from Geobacillus kaustophilus HTA426 was cloned, sequenced, and overexpressed into Escherichia coli Rosetta (DE3). The deduced protein was a 30-kDa monomeric esterase with high homology to carboxylesterases from Geobacillus thermoleovorans NY (99% identity) and Geobacillus stearothermophilus (97% identity). This protein suffered a proteolytic cut in E. coli, and the problem was overcome by introducing a mutation in the gene (K212R) without affecting the activity. The resulting Est30 showed remarkable thermostability at 65 degrees C, above the optimum growth temperature of G. kaustophilus HTA426. The optimum pH of the enzyme was 8.0. In addition, the purified enzyme exhibited stability against denaturing agents, like organic solvents, detergents, and urea. The protein catalyzed the hydrolysis of p nitrophenyl esters of different acyl chain lengths, confirming the esterase activity. The sequence analysis showed that the protein contains a catalytic triad formed by Ser93, Asp192, and His222, and the Ser of the active site is located in the conserved motif Gly91-X-Ser93-X-Gly95 included in most esterases and lipases. However, this carboxylesterase showed no more than 17% sequence identity with the closest members in the eight families of microbial carboxylesterases. The three-dimensional structure was modeled by sequence alignment and compared with others carboxylesterases. The topological differences suggested the classification of this enzyme and other Geobacillus-related carboxylesterases in a new alpha/beta hydrolase family different from IV and VI. PMID- 19304851 TI - Role of class A penicillin-binding proteins in the expression of beta-lactam resistance in Enterococcus faecium. AB - Peptidoglycan is polymerized by monofunctional d,d-transpeptidases belonging to class B penicillin-binding proteins (PBPs) and monofunctional glycosyltransferases and by bifunctional enzymes that combine both activities (class A PBPs). Three genes encoding putative class A PBPs (pbpF, pbpZ, and ponA) were deleted from the chromosome of Enterococcus faecium D344R in all possible combinations in order to identify the glycosyltransferases that cooperate with low-affinity class B Pbp5 for synthesis of peptidoglycan in the presence of beta lactam antibiotics. The viability of the triple mutant indicated that glycan strands can be polymerized independently from class A PBPs by an unknown glycosyltranferase. The susceptibility of the DeltapbpF DeltaponA mutant and triple mutants to extended spectrum cephalosporins (ceftriaxone and cefepime) identified either PbpF or PonA as essential partners of Pbp5 for peptidoglycan polymerization in the presence of the drugs. Mass spectrometry analysis of peptidoglycan structure showed that loss of PonA and PbpF activity led to a minor decrease in the extent of peptidoglycan cross-linking by the remaining PBPs without any detectable compensatory increase in the participation of the L,D transpeptidase in peptidoglycan synthesis. Optical density measurements and electron microscopy analyses showed that the DeltapbpF DeltaponA mutant underwent increased stationary-phase autolysis compared to the parental strain. Unexpectedly, deletion of the class A pbp genes revealed dissociation between the expression of resistance to cephalosporins and penicillins, although the production of Pbp5 was required for resistance to both classes of drugs. Thus, susceptibility of Pbp5-mediated peptidoglycan cross-linking to different beta lactam antibiotics differed as a function of its partner glycosyltransferase. PMID- 19304852 TI - An orthologue of Bacteroides fragilis NanH is the principal sialidase in Tannerella forsythia. AB - Sialidase activity is a putative virulence factor of the anaerobic periodontal pathogen Tannerella forsythia, but it is uncertain which genes encode this activity. Characterization of a putative sialidase, SiaHI, by others, indicated that this protein alone may not be responsible for all of the sialidase activity. We describe a second sialidase in T. forsythia (TF0035), an orthologue of Bacteroides fragilis NanH, and its expression in Escherichia coli. Sialidase activity of the expressed NanH was confirmed by using 2'-(4-methylumbelliferyl) alpha-D-N-acetylneuraminic acid as a substrate. Biochemical characterization of the recombinant T. forsythia NanH indicated that it was active over a broad pH range, with optimum activity at pH 5.5. This enzyme has high affinity for 2'-(4 methylumbelliferyl)-alpha-D-N-acetylneuraminic acid (K(m) of 32.9 +/- 10.3 microM) and rapidly releases 4-methylumbelliferone (V(max) of 170.8 +/- 11.8 nmol of 4-methylumbelliferone min(-1) mg of protein(-1)). E. coli lysates containing recombinant T. forsythia NanH cleave sialic acid from a range of substrates, with a preference for alpha2-3 glycosidic linkages. The genes adjacent to nanH encode proteins apparently involved in the metabolism of sialic acid, indicating that the NanH sialidase is likely to be involved in nutrient acquisition. PMID- 19304853 TI - Sialic acid (N-acetyl neuraminic acid) utilization by Bacteroides fragilis requires a novel N-acetyl mannosamine epimerase. AB - We characterized the nanLET operon in Bacteroides fragilis, whose products are required for the utilization of the sialic acid N-acetyl neuraminic acid (NANA) as a carbon and energy source. The first gene of the operon is nanL, which codes for an aldolase that cleaves NANA into N-acetyl mannosamine (manNAc) and pyruvate. The next gene, nanE, codes for a manNAc/N-acetylglucosamine (NAG) epimerase, which, intriguingly, possesses more similarity to eukaryotic renin binding proteins than to other bacterial NanE epimerase proteins. Unphosphorylated manNAc is the substrate of NanE, while ATP is a cofactor in the epimerase reaction. The third gene of the operon is nanT, which shows similarity to the major transporter facilitator superfamily and is most likely to be a NANA transporter. Deletion of any of these genes eliminates the ability of B. fragilis to grow on NANA. Although B. fragilis does not normally grow with manNAc as the sole carbon source, we isolated a B. fragilis mutant strain that can grow on this substrate, likely due to a mutation in a NAG transporter; both manNAc transport and NAG transport are affected in this strain. Deletion of the nanE epimerase gene or the rokA hexokinase gene, whose product phosphorylates NAG, in the manNAc enabled strain abolishes growth on manNAc. Thus, B. fragilis possesses a new pathway of NANA utilization, which we show is also found in other Bacteroides species. PMID- 19304854 TI - The glutathione/glutaredoxin system is essential for arsenate reduction in Synechocystis sp. strain PCC 6803. AB - Arsenic resistance in Synechocystis sp. strain PCC 6803 is mediated by an operon of three genes in which arsC codes for an arsenate reductase with unique characteristics. Here we describe the identification of two additional and nearly identical genes coding for arsenate reductases in Synechocystis sp. strain PCC 6803, which we have designed arsI1 and arsI2, and the biochemical characterization of both ArsC (arsenate reductase) and ArsI. Functional analysis of single, double, and triple mutants shows that both ArsI enzymes are active arsenate reductases but that their roles in arsenate resistance are essential only in the absence of ArsC. Based on its biochemical properties, ArsC belongs to a family that, though related to thioredoxin-dependent arsenate reductases, uses the glutathione/glutaredoxin system for reduction, whereas ArsI belongs to the previously known glutaredoxin-dependent family. We have also analyzed the role in arsenate resistance of the three glutaredoxins present in Synechocystis sp. strain PCC 6803 both in vitro and in vivo. Only the dithiolic glutaredoxins, GrxA (glutaredoxin A) and GrxB (glutaredoxin B), are able to donate electrons to both types of reductases in vitro, while GrxC (glutaredoxin C), a monothiolic glutaredoxin, is unable to donate electrons to either type. Analysis of glutaredoxin mutant strains revealed that only those lacking the grxA gene have impaired arsenic resistance. PMID- 19304855 TI - Genetic mapping of secretion and functional determinants of the Vibrio cholerae TcpF colonization factor. AB - Colonization of the human small intestine by Vibrio cholerae requires the type IV toxin-coregulated pilus (TCP). TcpF, which is encoded within the tcp operon, is secreted from the bacterial cell by the TCP apparatus and is also essential for colonization. Bacteria lacking tcpF are deficient in colonization, and anti-TcpF antibodies are protective in the infant mouse cholera model. In order to elucidate the regions of the protein that are required for secretion through the TCP apparatus and for its function in colonization, random mutagenesis of tcpF was performed. Analysis of these mutants suggests that multiple regions throughout the protein influence extracellular secretion and that determinants near the C terminus are important for the function of TcpF in colonization. The TcpF proteins of certain environmental V. cholerae isolates with 31% to 66% identity to pathogenic V. cholerae TcpF showed higher similarity in regions identified as secretion determinants but diverged in regions found to be important for colonization. These environmental TcpF proteins are secreted from the pathogenic strain; however, they do not mediate colonization in the infant mouse model. Here we provide genetic evidence pointing toward regions of TcpF that influence secretion, as well as regions that play an important role in in vivo colonization. PMID- 19304856 TI - Structure and complexity of a bacterial transcriptome. AB - Although gene expression has been studied in bacteria for decades, many aspects of the bacterial transcriptome remain poorly understood. Transcript structure, operon linkages, and information on absolute abundance all provide valuable insights into gene function and regulation, but none has ever been determined on a genome-wide scale for any bacterium. Indeed, these aspects of the prokaryotic transcriptome have been explored on a large scale in only a few instances, and consequently little is known about the absolute composition of the mRNA population within a bacterial cell. Here we report the use of a high-throughput sequencing-based approach in assembling the first comprehensive, single nucleotide resolution view of a bacterial transcriptome. We sampled the Bacillus anthracis transcriptome under a variety of growth conditions and showed that the data provide an accurate and high-resolution map of transcript start sites and operon structure throughout the genome. Further, the sequence data identified previously nonannotated regions with significant transcriptional activity and enhanced the accuracy of existing genome annotations. Finally, our data provide estimates of absolute transcript abundance and suggest that there is significant transcriptional heterogeneity within a clonal, synchronized bacterial population. Overall, our results offer an unprecedented view of gene expression and regulation in a bacterial cell. PMID- 19304857 TI - Searching for protein-protein interactions within the Bacillus subtilis spore coat. AB - The capability of endospores of Bacillus subtilis to withstand extreme environmental conditions is secured by several attributes. One of them, the protein shell that encases the spore and is known as the coat, provides the spore with its characteristic resistance to toxic chemicals, lytic enzymes, and predation by unicellular and multicellular eukaryotes. Despite most of the components of the spore coat having been identified, we have only a vague understanding of how such a complex structure is assembled. Using the yeast two hybrid system, we attempted to identify direct contacts among the proteins allocated to the insoluble fraction of the spore coat: CotV, CotW, CotX, CotY, and CotZ. We also examined whether they could interact with CotE, one of the most crucial morphogenetic proteins governing outer coat formation and also present in the insoluble fraction. Out of all 21 possible interactions we tested, 4 were found to be positive. Among these interactions, we confirmed the previous observation that CotE forms homo-oligomers. In addition, we observed homotypic interactions of CotY, strong interactions between CotZ and CotY, and relatively weak, yet significant, interactions between CotV and CotW. The results of this yeast two-hybrid analysis were confirmed by size exclusion chromatography of recombinant coat proteins and a pull-down assay. PMID- 19304858 TI - Clinical alertness to valproic acid-induced hyperammonemia--two case reports. PMID- 19304859 TI - Neuronal correlates of visual and auditory alertness in the DMT and ketamine model of psychosis. AB - Deficits in attentional functions belong to the core cognitive symptoms in schizophrenic patients. Alertness is a nonselective attention component that refers to a state of general readiness that improves stimulus processing and response initiation. The main goal of the present study was to investigate cerebral correlates of alertness in the human 5HT(2A) agonist and N-methyl-D aspartic acid (NMDA) antagonist model of psychosis. Fourteen healthy volunteers participated in a randomized double-blind, cross-over event-related functional magnetic resonance imaging (fMRI) study with dimethyltryptamine (DMT) and S ketamine. A target detection task with cued and uncued trials in both the visual and the auditory modality was used. Administration of DMT led to decreased blood oxygenation level-dependent response during performance of an alertness task, particularly in extrastriate regions during visual alerting and in temporal regions during auditory alerting. In general, the effects for the visual modality were more pronounced. In contrast, administration of S-ketamine led to increased cortical activation in the left insula and precentral gyrus in the auditory modality. The results of the present study might deliver more insight into potential differences and overlapping pathomechanisms in schizophrenia. These conclusions must remain preliminary and should be explored by further fMRI studies with schizophrenic patients performing modality-specific alertness tasks. PMID- 19304860 TI - Acute tryptophan depletion selectively attenuates cardiac slowing in an Eriksen flanker task. AB - In the present study, the effects of transiently lowering central serotonin levels by means of acute tryptophan depletion on measures of cognitive flexibility were examined. Flexible behaviour was measured in an Eriksen flanker task, and cardiac and electro-cortical responses to errors and congruent and incongruent stimuli were measured. The depletion was successful in lowering tryptophan levels and, as expected, it did not affect subjective mood. Depletion did not affect performance and electro-cortical measures and selectively affected cardiac measures. Depletion attenuated cardiac slowing to incongruent flanker stimuli but did not affect cardiac responses to congruent stimuli and errors. The selective effect on cardiac responses as compared to performance and electro cortical measures was in accordance with earlier findings, as well as the attenuation of cardiac slowing. The selective effect on the cardiac response to incongruent stimuli was unexpected. Detailed analyses showed a close connection to the earlier reported attenuation of the cardiac response to negative feedback, and the effect is explained in terms of reduced anticipation of the feedback stimulus due to enhanced punishment prediction. PMID- 19304861 TI - Timing of stroke in elderly people exposed to typical and atypical antipsychotics: a replication cohort study after the paper of Kleijer, et al. PMID- 19304862 TI - A single high dose of escitalopram increases mismatch negativity without affecting processing negativity or P300 amplitude in healthy volunteers. AB - Information processing deficits are commonly found in psychiatric illnesses, while at the biochemical level serotonin seems to play a role in nearly all psychiatric disorders. Processing negativity (PN), mismatch negativity (MMN) and P300 amplitude are electrophysiological measures of information processing. The present study was designed to replicate and further extent the results of our initial study on the effects of a low dose of escitalopram (10 mg) on MMN, PN and P300 amplitude. In a randomised, double-blind, cross-over experiment, 20 healthy male volunteers received either a single, orally administered dose of 15 mg escitalopram (a highly selective serotonin reuptake inhibitor (SSRI)) or placebo, after which their PN, MMN and P300 amplitude were assessed. Similar to our initial study with 10 mg escitalopram, 15 mg escitalopram significantly increased MMN, while it did not affect P300 amplitude. In contrast to our initial study, however, the currently higher dose of escitalopram did not increase PN. Results support the view that a broad range of increased serotonergic activity enhances MMN, while the relationship between serotonin and PN seems more complex. The current study does not support a serotonergic involvement in P300 amplitude. PMID- 19304863 TI - mCPP: an undesired addition to the ecstasy market. AB - A new ecstasy-like substance, meta-chlorophenylpiperazine (mCPP), has been detected in street drugs in the Netherlands. Theoretically, mCPP possesses the potential to become a non-neurotoxic alternative for methylenedioxymethamphetamine (MDMA), the regular psychoactive substance of ecstasy. Since its introduction on the Dutch market of synthetic drugs, the percentage of mCPP-containing tablets has increased, including both tablets that contain only mCPP and tablets containing a combination of mCPP and MDMA. These tablets occur in many different colours, shapes and sizes and with various logos, making it impossible to distinguish mCPP-containing tablets from regular MDMA tablets. In addition, the reports of users concerning the effects of mCPP are predominantly negative. All these aspects together lead to the conclusion that mCPP is an undesired addition to the ecstasy market from the user's perspective. PMID- 19304864 TI - An investigation of bupropion substitution for the interoceptive stimulus effects of nicotine. AB - Although the exact mechanism that makes bupropion hydrochloride (Zyban) effective as a smoking cessation aid has not been fully elucidated, studies have found that bupropion and nicotine share behavioural and neurophysiological properties suggesting that bupropion might serve as a substitute for nicotine. In fact, bupropion prompts nicotine-appropriate responding in operant and Pavlovian drug discrimination studies with rats. A majority of the literature examining this substitution pattern has been done with an operant paradigm. The present research extended this literature by further characterising the behavioural and neuropharmacological properties underlying the substitution for a nicotine conditioned stimulus (CS). Examination of the dose-effect function and temporal dynamics of this substitution pattern showed that bupropion (20 mg/kg) produced conditioned responding similar to nicotine (0.4 mg base/kg) (ED(50) = 9.9 mg/kg) at 15 and 30 min after injection and partially substituted 5 and 60 min post injection. Bupropion produced a pattern of conditioned responding similar to nicotine during a 60-min extinction test. Additionally, it has been hypothesised that bupropion and nicotine have an overlapping dopaminergic mechanism. We tested the effects of bupropion pretreatment, the nicotine dose-effect function and the ability of dopamine antagonist to block the substitution of bupropion for nicotine. Pretreatment with doses of bupropion that did not substitute for the nicotine stimulus (5 and 10 mg/kg) did not affect nicotine-conditioned responding; pretreatment with 20 mg/kg attenuated nicotine-evoked responding. Pretreatment with the dopamine antagonists SCH-23390 and eticlopride blocked the substitution. Finally, S,S-hydroxybupropion, the major metabolite of bupropion in humans, did not substitute for the nicotine CS. PMID- 19304865 TI - Are depot antipsychotics more coercive than tablets? The patient's perspective. AB - Some clinicians consider depot antipsychotics to be stigmatising, coercive and unacceptable to patients. This cross-sectional study investigated patients' perspectives of coercion for depot and oral antipsychotics. In all, 72 participants with chronic mental illness on voluntary maintenance antipsychotic medication were interviewed for their views on oral and depot medication and experiences of coercion. The MacArthur Admission Experience (short form) was adapted to explore coercion regarding medication. Mean total coercion levels were higher for those on depot (depot: mean 4.39; oral: 2.80, P = 0.027), as were perceived coercion (2.52 vs 1.73, P = 0.041) and negative pressures subscales (1.17 vs 0.33, P = 0.009). No significant differences were found for the 'voice' subscale and affective reactions. Specifically, more participants on depot felt that people try to force them to take medication (30% vs 2%, P< 0.001). Depots were perceived as more coercive than oral antipsychotics. Greater perceived coercion may explain why some consider depots to be a more stigmatising form of treatment. Although forced medication is sometimes required, the experience of coercion should be minimised by giving patients a fair say in treatment decisions, regardless of formulation. PMID- 19304866 TI - MDMA (ecstasy) use is associated with reduced BOLD signal change during semantic recognition in abstinent human polydrug users: a preliminary fMRI study. AB - 3,4-methylenedioxymethamphetamine (MDMA) users have impaired verbal memory, and voxel-based morphometry has shown decreased grey matter in Brodmann area (BA) 18, 21 and 45. Because these regions play a role in verbal memory, we hypothesized that MDMA users would show altered brain activation in these areas during performance of a functional magnetic resonance imaging (fMRI) task that probed semantic verbal memory. Polysubstance users enriched for MDMA exposure participated in a semantic memory encoding and recognition fMRI task that activated left BA 9, 18, 21/22 and 45. Primary outcomes were percent blood oxygen level-dependent signal change in left BA 9, 18, 21/22 and 45, accuracy and response time. During semantic recognition, lifetime MDMA use was associated with decreased activation in left BA 9, 18 and 21/22 but not 45. This was partly influenced by contributions from cannabis and cocaine use. MDMA exposure was not associated with accuracy or response time during the semantic recognition task. During semantic recognition, MDMA exposure was associated with reduced regional brain activation in regions mediating verbal memory. These findings partially overlap with previous structural evidence for reduced grey matter in MDMA users and may, in part, explain the consistent verbal memory impairments observed in other studies of MDMA users. PMID- 19304867 TI - Activation of the JAK-STAT pathway by olanzapine is necessary for desensitization of serotonin2A receptor-stimulated phospholipase C signaling in rat frontal cortex but not serotonin2A receptor-stimulated hormone release. AB - Chronic treatment with olanzapine causes desensitization of serotonin 2A receptor signaling. The purpose of the current study was to further understand the mechanisms underlying this desensitization response of serotonin 2A receptor signaling in vivo. We report that desensitization of serotonin 2A receptor stimulated-phospholipase C activity in rat frontal cortex induced by olanzapine is dependent on the activation of the JAK-STAT pathway. Olanzapine treatment for 7 days significantly increased the levels of the regulator of G protein signaling (RGS7) protein, RGS7 mRNA levels, and activation of JAK2 in rat frontal cortex. Pre-treatment with a JAK2 inhibitor AG490, significantly attenuated the olanzapine-induced reductions in serotonin 2A receptor-stimulated phospholipase C activity and prevented the olanzapine-induced increases in RGS7 mRNA and protein levels. In contrast, inhibition of the JAK-STAT pathway with AG490 did not reverse the olanzapine-induced desensitization of the serotonin 2A receptor pathway in the hypothalamic paraventricular nucleus mediating increases in plasma hormone levels. AG490 dose-dependently inhibited serotonin 2A receptor-stimulated oxytocin and corticosterone release. These results suggest that the olanzapine induced increase in RGS7 expression is mediated by the activation of JAK-STAT and is necessary for olanzapine-induced desensitization of serotonin 2A receptor stimulated phospholipase C activity in the frontal cortex but not serotonin 2A receptor-stimulated hormone release. PMID- 19304868 TI - The prevalence of undiagnosed metabolic abnormalities in people with serious mental illness. AB - The prevalence of metabolic syndrome is increased 2-3-fold in people with serious mental illness (SMI). Monitoring of physical health in these individuals is poor, despite clear guidance from the National Institute of Health and Clinical Excellence. The aim of this study was to assess the proportion of people with SMI who had been screened for metabolic abnormalities within the previous year and in a further study to assess the prevalence of undiagnosed metabolic abnormalities in people who had not been screened. The notes and computer records of 100 patients with SMI from community and in-patient settings were evaluated. In a subsequent study, the prevalence of metabolic syndrome was assessed in 71 previously unscreened patients. The study was carried out at the psychiatric in patient and out-patient units in Southampton and Winchester. The frequency of screening and prevalence of the metabolic syndrome as defined by the International Diabetes Federation (IDF) were assessed. There was documented evidence that the following cardiovascular risk factors had been measured in the previous year: blood pressure (32%), glucose (16%), lipids (9%) and weight (2%). In the metabolic abnormalities study, 41 of 71 (58%) patients were found to fulfil the IDF criteria for the metabolic syndrome. Two had previously undiagnosed diabetes. Twelve percent of patients had a greater than 20% risk of a cardiovascular event within the next 10 years. Despite clear guidance and a high prevalence of undiagnosed metabolic syndrome, screening rates for metabolic abnormalities in people with SMI remain low. Improved screening of metabolic complications should lead to better identification and treatment of this clinical problem. PMID- 19304869 TI - Association of the IGF1/pregnancy-associated plasma protein-A system and adipocytokine levels with the presence and the morphology of carotid plaques in type 2 diabetes mellitus patients with stable glycaemic control. AB - OBJECTIVE: Pregnancy-associated plasma protein-A (PAPP-A) has been implicated in the atherosclerotic process through regulation of local expression of IGF1. In type 2 diabetes mellitus, glycaemic control has been involved in PAPP-A expression. We compared PAPP-A, IGF1, inflammatory markers and adiponectin concentrations in type 2 diabetic patients with and without carotid plaques and evaluated the relationship between these serum parameters and ultrasound carotid markers of atherosclerosis. METHODS: We studied 125 consecutive type 2 diabetic patients. Clinical data, metabolic variables, hemostatic factors (plasma type-1 plasminogen activator inhibitor, fibrinogen), high-ultrasensitive C reactive protein (hsCRP), tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, adiponectin, IGF1 and PAPP-A were determined. Patients were classified into two groups according to the presence of carotid plaques on ultrasound. Carotid intima media thickness (IMT) and morphology of carotid plaques were evaluated. RESULTS: The mean age was 61.5+/-7.3 years and the mean glycated hemoglobin of 6.8+/-0.9%. A total of 60% presented carotid plaques. Both groups were homogeneous in anthropometric data, biochemical determinations and hemostatic factors. Adiponectin, hsCRP, TNF-alpha and IL-6 were similar in both groups. No differences were observed in serum PAPP-A (0.46 (0.22-0.86) vs 0.38 (0.18-0.66) mIU/l and in SDS IGF1 (-0.34+/-1.38 vs -0.67+/-1.35)) in patients with and without carotid plaques respectively. PAPP-A and IGF1 were not correlated with IMT. CONCLUSIONS: Serum PAPP-A and IGF1 do not appear to be useful serum biomarkers for carotid atherosclerosis in type 2 diabetic patients with stable glycemic control, despite scientific evidence of their local role in atherosclerosis. PMID- 19304870 TI - The role of estrogens for male bone health. AB - Sex steroids are important for the growth and maintenance of both the female and the male skeleton. However, the relative contribution of androgens versus estrogens in the regulation of the male skeleton is unclear. Experiments using mice with inactivated sex steroid receptors demonstrated that both activation of the estrogen receptor (ER)alpha and activation of the androgen receptor result in a stimulatory effect on both the cortical and trabecular bone mass in males. ERbeta is of no importance for the skeleton in male mice while it modulates the ERalpha-action on bone in female mice. Previous in vitro studies suggest that the membrane G protein-coupled receptor GPR30 also might be a functional ER. Our in vivo analyses of GPR30-inactivated mice revealed no function of GPR30 for estrogen-mediated effects on bone mass but it is required for normal regulation of the growth plate and estrogen-mediated insulin-secretion. Recent clinical evidence suggests that a threshold exists for estrogen effects on bone in men: rates of bone loss and fracture risk seem to be the highest in men with estradiol levels below this threshold. Taken together, even though these findings do not exclude an important role for testosterone in male skeletal homeostasis, it is now well-established that estrogens are important regulators of bone health in men. PMID- 19304871 TI - Semantic Web for Health Care and Life Sciences: a review of the state of the art. PMID- 19304872 TI - Scaling the walls of discovery: using semantic metadata for integrative problem solving. AB - Current data integration approaches by bioinformaticians frequently involve extracting data from a wide variety of public and private data repositories, each with a unique vocabulary and schema, via scripts. These separate data sets must then be normalized through the tedious and lengthy process of resolving naming differences and collecting information into a single view. Attempts to consolidate such diverse data using data warehouses or federated queries add significant complexity and have shown limitations in flexibility. The alternative of complete semantic integration of data requires a massive, sustained effort in mapping data types and maintaining ontologies. We focused instead on creating a data architecture that leverages semantic mapping of experimental metadata, to support the rapid prototyping of scientific discovery applications with the twin goals of reducing architectural complexity while still leveraging semantic technologies to provide flexibility, efficiency and more fully characterized data relationships. A metadata ontology was developed to describe our discovery process. A metadata repository was then created by mapping metadata from existing data sources into this ontology, generating RDF triples to describe the entities. Finally an interface to the repository was designed which provided not only search and browse capabilities but complex query templates that aggregate data from both RDF and RDBMS sources. We describe how this approach (i) allows scientists to discover and link relevant data across diverse data sources and (ii) provides a platform for development of integrative informatics applications. PMID- 19304873 TI - Semantic web for integrated network analysis in biomedicine. AB - The Semantic Web technology enables integration of heterogeneous data on the World Wide Web by making the semantics of data explicit through formal ontologies. In this article, we survey the feasibility and state of the art of utilizing the Semantic Web technology to represent, integrate and analyze the knowledge in various biomedical networks. We introduce a new conceptual framework, semantic graph mining, to enable researchers to integrate graph mining with ontology reasoning in network data analysis. Through four case studies, we demonstrate how semantic graph mining can be applied to the analysis of disease causal genes, Gene Ontology category cross-talks, drug efficacy analysis and herb drug interactions analysis. PMID- 19304874 TI - Automated diagnosis of LC-MS/MS performance. AB - SUMMARY: We report a software scheme for automated diagnosis of liquid chromatography tandem mass spectrometry (LC-MS/MS) system performance. The proposed software scheme provides a robust framework for establishing automated diagnosis of LC-MS/MS system performance for a variety of instruments and experiments. This schematic consists of four main software components: (i) data conversion, (ii) peptide identification, (iii) LC retention time analysis and (iv) system performance evaluation. The implementation of a standard approach for assessing LC-MS/MS system performance enables researchers to apply reliable metrics to assess their workflows performance over different batch experiments. Furthermore, the results from system diagnosis can provide feedback to the workflow to stop batch analysis if system performance falls below prescribed thresholds. A basic implementation of the approach based on the MassMatrix database search and LC retention time analysis programs is presented. AVAILABILITY: An open source implementation of the LC-MS/MS system diagnosis software based on the MassMatrix database search program is freely available to non-commercial users and can be downloaded at www.massmatrix.net. PMID- 19304875 TI - Local RNA structure alignment with incomplete sequence. AB - MOTIVATION: Accuracy of automated structural RNA alignment is improved by using models that consider not only primary sequence but also secondary structure information. However, current RNA structural alignment approaches tend to perform poorly on incomplete sequence fragments, such as single reads from metagenomic environmental surveys, because nucleotides that are expected to be base paired are missing. RESULTS: We present a local RNA structural alignment algorithm, trCYK, for aligning and scoring incomplete sequences under a model using primary sequence conservation and secondary structure information when possible. The trCYK algorithm improves alignment accuracy and coverage of sequence fragments of structural RNAs in simulated metagenomic shotgun datasets. AVAILABILITY: The source code for Infernal 1.0, which includes trCYK, is available at http://infernal.janelia.org. PMID- 19304876 TI - The calibrated population resistance tool: standardized genotypic estimation of transmitted HIV-1 drug resistance. AB - The calibrated population resistance (CPR) tool is a web-accessible program for performing standardized genotypic estimation of transmitted HIV-1 drug resistance. The program is linked to the Stanford HIV drug resistance database and can additionally perform viral genotyping and algorithmic estimation of resistance to specific antiretroviral drugs. AVAILABILITY: http://cpr.stanford.edu/cpr/index.html. PMID- 19304877 TI - GNU MCSim: Bayesian statistical inference for SBML-coded systems biology models. AB - SUMMARY: Statistical inference about the parameter values of complex models, such as the ones routinely developed in systems biology, is efficiently performed through Bayesian numerical techniques. In that framework, prior information and multiple levels of uncertainty can be seamlessly integrated. GNU MCSim was precisely developed to achieve those aims, in a general non-linear differential context. Starting with version 5.3.0, GNU MCSim reads in and simulates Systems Biology Markup Language models. Markov chain Monte Carlo simulations can be used to generate samples from the joint posterior distribution of the model parameters, given a dataset and prior distributions. Hierarchical statistical models can be used. Optimal design of experiments can also be investigated. AVAILABILITY AND IMPLEMENTATION: The GNU GPL source is available at (http://savannah.gnu.org/projects/mcsim). A distribution package is at (http://www.gnu.org/software/mcsim). GNU MCSim is written in standard C and runs on any platform supporting a C compiler. Supplementary Material is available online at (http://www.gnu.org/software/mcsim). PMID- 19304878 TI - Biopython: freely available Python tools for computational molecular biology and bioinformatics. AB - SUMMARY: The Biopython project is a mature open source international collaboration of volunteer developers, providing Python libraries for a wide range of bioinformatics problems. Biopython includes modules for reading and writing different sequence file formats and multiple sequence alignments, dealing with 3D macro molecular structures, interacting with common tools such as BLAST, ClustalW and EMBOSS, accessing key online databases, as well as providing numerical methods for statistical learning. AVAILABILITY: Biopython is freely available, with documentation and source code at (www.biopython.org) under the Biopython license. PMID- 19304879 TI - Cisplatin preparation error; patient management and morbidity. AB - INTRODUCTION: Antineoplastic drug therapy errors represent a high iatrogenic potential due to antineoplastic drugs narrow therapeutic ranges and the complexity of chemotherapy regimens that may increase the risk of morbidity and mortality for oncology patients. SETTING: We report a 57-year-old man with head and neck cancer who mistakenly received 180 mg/ m(2) of cisplatin overdose despite the safety measures and validations carried out during preparation. The patient developed moderate nausea and vomiting, acute renal failure, hearing difficulty (tinnitus), and severe myelodepression. PATIENT MANAGEMENT: Prophylactic and symptomatic treatments were applied in order to prevent and correct toxicity during the 9 days stay at hospital. RESULT: He recovered with mild tinnitus and mild renal impairment as the only sequelae. This case presents a hospital stay and treatment quite different to others used to reverse all cisplatin overdose toxicity and it shows the benefits of prompt management. PMID- 19304880 TI - Pharmacokinetics of a granisetron transdermal system for the treatment of chemotherapy-induced nausea and vomiting. AB - OBJECTIVE: To determine the pharmacokinetic (PK) profile of granisetron transdermal formulation and examine its possible relationship with age, gender, and renal function. METHODS: This article describes a Phase I PK study and a post hoc pooled population PK analysis. The Phase I study was a randomized, cross-over study that assessed PK parameters of three granisetron patch sizes and oral granisetron. The pooled population PK analysis included data from three trials in healthy subjects (n = 48) and from Phase II and III studies in patients with cancer (n = 793). The population PK model was used to investigate granisetron exposure and its possible relationship with age, gender, and renal function. RESULTS: Following oral dosing, plasma granisetron concentration was quantifiable at 1 h, and maximal mean concentration (4.7 ng/mL) was reached 2 h after administration. With transdermal application, maximal concentration was reached 48 h post-application; t(1/2) was 36 h. With oral dosing, overall exposure after 5 days was 306 ng/mL.h, and C(avg) 2.6 ng/mL. This corresponded to an AUC(0 infinity) for the 52 cm(2) patch of 420 ng/mL.h and C(avg) 2.2 ng/mL over 6 days. Clearance was not affected by age, gender, weight, or renal function. CONCLUSION: The 52 cm( 2) granisetron patch achieves a similar exposure to that of a 2 mg oral dose and provides continuous delivery of granisetron over 6 days. The patch may have utility in treating chemotherapy-induced nausea and vomiting where prolonged drug delivery is advantageous. No dose adjustments would be needed based on age or renal function. PMID- 19304881 TI - Influence of plasma exchange on the disposition of the fourth generation cephalosporin cefepime. AB - Cefepime, a fourth generation cephalosporin, is widely used in hematology and oncology patients. These patients may require plasma exchange (PE) for indications such as chemotherapy- or cancer-induced thromobotic thrombocytopenic purpura to name a few. To date, no pharmacokinetic evaluation has been conducted assessing cefepime's disposition during PE. A 2 g IV cefepime single dose was given to patients undergoing therapeutic PE. Two hours from cefepime dose administration, plasma concentration was measured. PE was then instituted and cefepime plasmapheresate concentration was measured at the completion of the PE session. Cefepime levels were measured using HPLC. The percentage removed by PE was calculated as: amount removed/2 g dose. Ten adult patients were analyzed: median age (range): 52 years (33-67) and median weight (range); 82.85 kg (47 120). PE indications were: myasthenia gravis (n = 3), transverse myelitis (n = 2), multiple sclerosis (n = 1), chronic inflammatory demyelinating polyneuropathy (n = 1), idiopathic thrombocytopenic purpura (n = 1), thrombotic thrombocytopenic purpura (n = 1), and humoral rejection post cadaveric renal allograft (n = 1). All patients except one had a creatinine clearance >60 mL/min. One patient was excluded from the pharmacokinetic analysis owing to loss of venous access during PE. For the remaining nine patients, total plasma volume removed was 3.5 L (range: 2.5-3.5) and duration of PE was 120 min (range: 94-209). The cefepime removed by PE was 3.7% (range: 2.1-6.7). A strong correlation was found between cefepime plasma concentration prior to PE and the amount of drug removed (r = 0.96, r(2) = 0.92; p<0.05). The above results suggest that, under the studied conditions, cefepime removal by PE is clinically insignificant (approximately 4% of total 2 g dose). PMID- 19304882 TI - Cytotoxic chemotherapy for pregnancy-associated breast cancer: single institution case series. AB - BACKGROUND: Pregnancy-associated (PA) breast cancer is a rare disease state that poses unique management challenges, specifically controlling the cancer and maximizing the survival of the expectant mother balanced with the health and safety of the developing fetus. As more women delay pregnancy into their 30s and 40s it is expected that this may become a more important clinical problem in the future. Existing data on PA-breast cancer comes from case series using older chemotherapy drugs. A review of practice was carried out to assess current experience with PA-breast cancer, particularly relating to current cytotoxic drugs and targeted agents. METHODS: The St James's Hospital breast cancer registry, a prospectively maintained database, was used to identify cases of PA breast cancer over a 6.5-year period and a chart review carried out. Chemotherapy administered during pregnancy, breast cancer specific outcomes, and fetal outcomes were assessed. RESULTS: Five patients were identified with PA-breast cancer; median age 34 years (range 28-35). The median gestation at presentation was 18 weeks (range 14-29). Four women received chemotherapy during pregnancy; three received doxorubicin and cyclophosphamide (AC) and one paclitaxel. These agents were generally well tolerated. At median gestation of 36 weeks (range 35 40 weeks) four elective caesareans and one spontaneous delivery occurred. There were no fetal abnormalities. CONCLUSIONS: Common cytotoxics can safely be delivered in pregnancy. Further research on newer therapies such as trastuzumab is needed. PMID- 19304883 TI - Interpretation of medical information acts by UK occupational physicians. AB - BACKGROUND: Difficulties arise in applying the Data Protection Act 1998 and the Access to Medical Reports Act 1988 in occupational health practice. There is no guidance on detailed aspects of applying these Acts in practice and consistent advice has proved difficult to obtain. AIMS: To audit the understanding and practice of UK occupational physicians to see if a consensus view existed. METHODS: A postal questionnaire sent to all UK-based Society of Occupational Medicine (SOM) members between December 2005 and June 2006. Responses were analysed using the SPSS 13.0 software. RESULTS: Responses were received from 726 SOM members, a response rate of 48%. The study revealed wide variation and a limited consensus in practice. Significant differences existed between doctors with a Diploma in Occupational Medicine and those with higher Faculty qualifications, between part-time and full-time practitioners and between doctors who qualified pre- and post-1974. CONCLUSIONS: The audit revealed wide variation in responding to clinical scenarios in relation to both the Access to Medical Reports and the Data Protection Acts. The findings have implications for clinical practice, policy and research. The majority of respondents reported that national guidance is needed. PMID- 19304884 TI - Factors associated with farmers joining occupational health services. AB - BACKGROUND: Agriculture is one of the most hazardous occupations in the world. The aim of an occupational health service (OHS) is to reduce the risk of work related diseases, occupational diseases and injuries and to promote workers' health. In Finland, they are part of the public health care system, but for farmers OHS is voluntary. AIMS: To explore factors associated with farmers joining farmers' occupational health services (FOHS). This knowledge is important for improving the coverage of FOHS and to motivate farmers to join. METHODS: In 2004 and 2005, we interviewed a total of 1182 full-time farmers aged 18-64, chosen randomly from the register of the Information Centre of the Ministry of Agricultural and Forestry in Finland (Tike). A stepwise multivariate logistic regression model was used to calculate odds ratios and their 95% confidence intervals for factors associated with their joining FOHS. RESULTS: Among both genders, chronic illnesses, farm size and opinion on whether membership of FOHS should be obligatory were predictors of farmers joining FOHS. Among male farmers, the production sector, the existence of a quality system on the farm and vocational education were associated with interest in joining. Among female farmers, interest was associated with physical activity. CONCLUSIONS: Emphasizing FOHS constitutes a preventive approach. All farmers should be motivated by FOHS to become aware of issues concerning safety and health at work. The potential channels for increasing its awareness are vocational education, quality systems and support from farmers who are already members. PMID- 19304885 TI - Chemokine signaling in embryonic cell migration: a fisheye view. AB - Chemokines and their receptors were discovered about twenty years ago as mediators of leukocyte traffic. Over the past decade, functional studies of these molecules have revealed their importance for cell migration processes during embryogenesis, which, in addition to providing mechanistic insights into embryonic development, could complement information about chemokine function in the immune system. Here, we review the roles of the chemokine stromal cell derived factor 1 (SDF-1/CXCL12) and its receptor CXCR4 during zebrafish and mouse embryonic development, and discuss their function in regulating the interactions of cells with their extracellular environment, in directing their migration, and in maintaining their location. PMID- 19304886 TI - Neuronal activity and Wnt signaling act through Gsk3-beta to regulate axonal integrity in mature Drosophila olfactory sensory neurons. AB - The roles played by signaling pathways and neural activity during the development of circuits have been studied in several different contexts. However, the mechanisms involved in maintaining neuronal integrity once circuits are established are less well understood, despite their potential relevance to neurodegeneration. We demonstrate that maintenance of adult Drosophila olfactory sensory neurons requires cell-autonomous neuronal activity. When activity is silenced, development occurs normally, but neurons degenerate in adulthood. These detrimental effects can be compensated by downregulating Glycogen synthase kinase 3beta (Gsk-3beta). Conversely, ectopic expression of activated Gsk-3beta or downregulation of Wnt effectors also affect neuron stability, demonstrating a role for Wnt signaling in neuroprotection. This is supported by our observation that activated adult neurons are capable of increased Wingless release, and its targeted expression can protect neurons against degeneration. The role of Wnt signaling in this process is non-transcriptional, and may act on cellular mechanisms that regulate axonal or synaptic stability. Together, we provide evidence that Gsk-3beta is a key sensor involved in neural circuit integrity, maintaining axon stability through neural activity and the Wnt pathway. PMID- 19304887 TI - Recruitment and maintenance of tendon progenitors by TGFbeta signaling are essential for tendon formation. AB - Tendons and ligaments mediate the attachment of muscle to bone and of bone to bone to provide connectivity and structural integrity in the musculoskeletal system. We show that TGFbeta signaling plays a major role in the formation of these tissues. TGFbeta signaling is a potent inducer of the tendon progenitor (TNP) marker scleraxis both in organ culture and in cultured cells, and disruption of TGFbeta signaling in Tgfb2(-/-);Tgfb3(-/-) double mutant embryos or through inactivation of the type II TGFbeta receptor (TGFBR2; also known as TbetaRII) results in the loss of most tendons and ligaments in the limbs, trunk, tail and head. The induction of scleraxis-expressing TNPs is not affected in mutant embryos and the tendon phenotype is first manifested at E12.5, a developmental stage in which TNPs are positioned between the differentiating muscles and cartilage, and in which Tgfb2 or Tgfb3 is expressed both in TNPs and in the differentiating muscles and cartilage. TGFbeta signaling is thus essential for maintenance of TNPs, and we propose that it also mediates the recruitment of new tendon cells by differentiating muscles and cartilage to establish the connections between tendon primordia and their respective musculoskeletal counterparts, leading to the formation of an interconnected and functionally integrated musculoskeletal system. PMID- 19304888 TI - Map2k1 and Map2k2 genes contribute to the normal development of syncytiotrophoblasts during placentation. AB - The mammalian genome contains two ERK/MAP kinase kinase genes, Map2k1 and Map2k2, which encode dual-specificity kinases responsible for ERK/MAP kinase activation. In the mouse, loss of Map2k1 function causes embryonic lethality, whereas Map2k2 mutants survive with a normal lifespan, suggesting that Map2k1 masks the phenotype due to the Map2k2 mutation. To uncover the specific function of MAP2K2 and the threshold requirement of MAP2K proteins during embryo formation, we have successively ablated the Map2k gene functions. We report here that Map2k2 haploinsufficiency affects the normal development of placenta in the absence of one Map2k1 allele. Most Map2k1(+/-)Map2k2(+/-) embryos die during gestation because of placenta defects restricted to extra-embryonic tissues. The impaired viability of Map2k1(+/-)Map2k2(+/-) embryos can be rescued when the Map2k1 deletion is restricted to the embryonic tissues. The severity of the placenta phenotype is dependent on the number of Map2k mutant alleles, the deletion of the Map2k1 allele being more deleterious. Moreover, the deletion of one or both Map2k2 alleles in the context of one null Map2k1 allele leads to the formation of multinucleated trophoblast giant (MTG) cells. Genetic experiments indicate that these structures are derived from Gcm1-expressing syncytiotrophoblasts (SynT), which are affected in their ability to form the uniform SynT layer II lining the maternal sinuses. Thus, even though Map2k1 plays a predominant role, these results enlighten the function of Map2k2 in placenta development. PMID- 19304889 TI - LIM homeobox transcription factors integrate signaling events that control three dimensional limb patterning and growth. AB - Vertebrate limb development is controlled by three signaling centers that regulate limb patterning and growth along the proximodistal (PD), anteroposterior (AP) and dorsoventral (DV) limb axes. Coordination of limb development along these three axes is achieved by interactions and feedback loops involving the secreted signaling molecules that mediate the activities of these signaling centers. However, it is unknown how these signaling interactions are processed in the responding cells. We have found that distinct LIM homeodomain transcription factors, encoded by the LIM homeobox (LIM-HD) genes Lhx2, Lhx9 and Lmx1b integrate the signaling events that link limb patterning and outgrowth along all three axes. Simultaneous loss of Lhx2 and Lhx9 function resulted in patterning and growth defects along the AP and the PD limb axes. Similar, but more severe, phenotypes were observed when the activities of all three factors, Lmx1b, Lhx2 and Lhx9, were significantly reduced by removing their obligatory co-factor Ldb1. This reveals that the dorsal limb-specific factor Lmx1b can partially compensate for the function of Lhx2 and Lhx9 in regulating AP and PD limb patterning and outgrowth. We further showed that Lhx2 and Lhx9 can fully substitute for each other, and that Lmx1b is partially redundant, in controlling the production of output signals in mesenchymal cells in response to Fgf8 and Shh signaling. Our results indicate that several distinct LIM-HD transcription factors in conjunction with their Ldb1 co-factor serve as common central integrators of distinct signaling interactions and feedback loops to coordinate limb patterning and outgrowth along the PD, AP and DV axes after limb bud formation. PMID- 19304890 TI - Sonic hedgehog signaling regulates reciprocal epithelial-mesenchymal interactions controlling palatal outgrowth. AB - The mammalian secondary palate arises by outgrowth from the oral side of the paired maxillary processes flanking the primitive oral cavity. Palatal growth depends on reciprocal interactions between the oral ectoderm and the underlying neural-crest-derived mesenchyme. Previous studies have implicated sonic hedgehog (Shh) as an important epithelial signal for regulating palatal growth. However, the cellular and molecular mechanisms through which Shh regulates palatal development in vivo have not been directly analyzed, due in part to early embryonic lethality of mice lacking Shh or other essential components of the Shh signaling pathway. Using Cre/loxP-mediated tissue-specific inactivation of the smoothened (Smo) gene in the developing palatal mesenchyme, we show that the epithelially expressed Shh signals directly to the palatal mesenchyme to regulate palatal mesenchyme cell proliferation through maintenance of cyclin D1 (Ccnd1) and Ccnd2 expression. Moreover, we show that Shh-Smo signaling specifically regulates the expression of the transcription factors Foxf1a, Foxf2 and Osr2 in the developing palatal mesenchyme. Furthermore, we show that Shh signaling regulates Bmp2, Bmp4 and Fgf10 expression in the developing palatal mesenchyme and that specific inactivation of Smo in the palatal mesenchyme indirectly affects palatal epithelial cell proliferation. Together with previous reports that the mesenchymally expressed Fgf10 signals to the palatal epithelium to regulate Shh mRNA expression and cell proliferation, these data demonstrate that Shh signaling plays a central role in coordinating the reciprocal epithelial mesenchymal interactions controlling palatal outgrowth. PMID- 19304891 TI - An oral human drug absorption study to assess the impact of site of delivery on the bioavailability of bevirimat. AB - Bevirimat is a first in class, orally active, potent and selective inhibitor of HIV-1. It may have utility for the treatment of HIV-1-infected patients who are failing current regimens because of the development of drug resistance. In earlier studies in HIV-1-infected patients, an immediate-release tablet formulation exhibited a relative bioavailability (F(rel)) of 50% or greater and a higher intersubject variability than an oral solution. The purpose of this study was to determine whether this pharmacokinetic profile is attributable to a narrow permeability-dependent absorption window within the gastrointestinal tract. Three groups of subjects completed an open-label, 2-way crossover, randomized pharmacoscintigraphic study. Subjects received a 25-mg bevirimat oral immediate release solution plus 25 mg bevirimat solution delivered to the proximal small bowel, distal small bowel, or colon via the Enterion capsule. The results indicate that the permeability of bevirimat throughout the small intestine was excellent and suggest that the variability observed for the immediate release tablet was not related to the presence of an absorption window in the small intestine. PMID- 19304892 TI - Storage or retrieval deficit: the yin and yang of amnesia. AB - To this day, it remains unresolved whether experimental amnesia reflects failed memory storage or the inability to retrieve otherwise intact memory. Methodological as well as conceptual reasons prevented deciding between these two alternatives: The absence of recovery from amnesia is typically taken as supporting storage impairment interpretations; however, this absence of recovery does not positively demonstrate nonexistence of memory, allowing for alternative interpretations of amnesia as impairment of memory retrieval. To address this shortcoming, we present a novel approach to study the nature of amnesia that makes positive, i.e., falsifiable, predictions for the absence of memory. Applying this paradigm, we demonstrate here that infusing anisomycin into the dorsal hippocampus induces amnesia by impairing memory storage, not retrieval. PMID- 19304893 TI - The hippocampus supports encoding of between-domain associations within working memory. AB - It has been established that the medial temporal lobe, including the hippocampus, is crucial for associative memory. The aim of the current functional magnetic resonance imaging (fMRI) study was to investigate whether the hippocampus is differentially activated for associations between items processed in the same neocortical region (within-domain) as compared with associations between items processed in different neocortical regions (between-domain). Here, we show that the hippocampus is significantly more active for between-domain associations compared with within-domain associations. Thus, the hippocampus is important for associative encoding, and furthermore, shows greater activation when the stimuli to be associated come from different stimulus categories. PMID- 19304894 TI - The basolateral amygdala is necessary for the encoding and the expression of odor memory. AB - Conditioned odor avoidance (COA) results from the association between a novel odor and a delayed visceral illness. The present experiments investigated the role of the basolateral amygdala (BLA) in acquisition and retrieval of COA memory. To address this, we used the GABA(A) agonist muscimol to temporarily inactivate the BLA during COA acquisition or expression. BLA inactivation before odor-malaise pairing greatly impaired COA tested 3 d later. In contrast, muscimol microinfusion between odor and malaise spared retention. Moreover, inactivation of the BLA before pre-exposure to the odor prevented latent inhibition of COA. This suggests that neural activity in the BLA is essential for the formation of odor representation. BLA inactivation before the retrieval test also blocked COA memory expression when performed either 3 d (recent memory) or 28 d (remote memory) after acquisition. This effect was transitory as muscimol-treated animals were not different from controls during the subsequent extinction tests. Moreover, muscimol infusion in the BLA neither affected olfactory perception nor avoidance behavior, and it did not induce a state-dependent learning. Altogether, these findings suggest that neural activity in the BLA is required for the encoding and the retrieval of odor memory. Moreover, the BLA seems to play a permanent role in the expression of COA. PMID- 19304895 TI - Interferon-lambdas: the modulators of antivirus, antitumor, and immune responses. AB - IFN-lambdas, including IFN-lambda1, IFN-lambda2, and IFN-lambda3, also known as IL-29, IL-28A, or IL-28B, are a newly described group of cytokines distantly related to the type I IFNs and IL-10 family members. The IFN-lambdaR complex consists of a unique ligand-binding chain, IFN-lambdaR1 (also designated IL 28Ralpha), and an accessory chain, IL-10R2, which is shared with receptors for IL 10-related cytokines. IFN-lambdas signal through the IFN-lambdaR and activate pathways of JAK-STATs and MAPKs to induce antiviral, antiproliferative, antitumor, and immune responses. In this review, we summarize recent findings about the biology of IFN-lambdas and their pathophysiological roles in viral infection, cancer, and immune responses of the innate and adaptive arms. PMID- 19304896 TI - Myeloid blasts are the mouse bone marrow cells prone to differentiate into osteoclasts. AB - Cells of the myeloid lineage at various stages of maturity can differentiate into multinucleated osteoclasts. Yet, it is unclear which developmental stages of this lineage are more prone to become osteoclasts than others. We investigated the osteoclastogenic potential of three successive stages of myeloid development isolated from mouse bone marrow. Early blasts (CD31hi/Ly-6C-), myeloid blasts (CD31+/Ly-6C+), and monocytes (CD31-/Ly-6Chi), as well as unfractionated marrow cells, were cultured in the presence of M-CSF and receptor activator of NF-B ligand (RANKL), and the differentiation toward multinucleated cells and their capacity to resorb bone was assessed. Myeloid blasts developed rapidly into multinucleated cells; in only 4 days, maximal numbers were reached, whereas the other fractions required 8 days to reach maximal numbers. Bone resorption was observed after 6 (myeloid blasts and monocyte-derived osteoclasts) and 8 (early blast-derived osteoclasts) days. This difference in kinetics in osteoclast forming capacity was confirmed by the analysis of osteoclast-related genes. In addition, the myeloid blast fraction proved to be most sensitive to M-CSF and RANKL, as assessed with a colony-forming assay. Our results show that osteoclasts can develop from all stages of myeloid differentiation, but myeloid blasts are equipped to do so within a short period of time. PMID- 19304897 TI - Diagnosis and management of otalgia in children. PMID- 19304898 TI - Paediatric trauma: injury pattern and mortality in the UK. AB - OBJECTIVE: Trauma accounts for a large proportion of childhood deaths. No data exist about injury patterns within paediatric trauma in the UK. Identification of specific high-risk injury patterns may lead to improved care and outcome. METHODS: Data from 24 218 paediatric trauma cases recorded by the Trauma Audit and Research Network (TARN) from 1990 to 2005 were analysed. Main injury, injury patterns and outcome were analysed. Mortality at 93 days' post-injury was the major outcome measure. RESULTS: Limb injuries occurred in 65.0% of patients. In infants 81.4% of head injuries were isolated, compared with 46.5% in 11-15-year old children. Thoracic injuries were associated with other injuries in 68.4%. The overall mortality rate was 3.7% (n = 893). Mortality decreased from 4.2% to 3.1%; this was most evident in non-isolated head injuries. It was low in isolated injuries: 1.5% (n = 293). In children aged 1-15 years the highest mortalities occurred in multiple injuries including head/thoracic (47.7%) and head/abdominal injuries (49.9%). Having a Glasgow Coma Scale of <15 on presentation to hospital was associated with a mortality of 16%. CONCLUSIONS: Differences in injury patterns and mortality exist between different age groups and high-risk injury patterns can be identified. With increasing age, a decline in the proportion of children with head injury and an increase in the proportion with limb injury were observed. This information is useful for directing ongoing care of severely injured children. Future analyses of the TARN database may help to evaluate the management of high-risk children and to identify the most effective care. PMID- 19304899 TI - Age no barrier to diagnosis. PMID- 19304900 TI - Guideline review: management of invasive meningococcal disease, SIGN. PMID- 19304901 TI - The many faces of cellulitis. PMID- 19304902 TI - Prescribing in paediatrics. PMID- 19304904 TI - Group interpersonal therapy reduces depression in adolescent survivors of war. PMID- 19304905 TI - Review: Psychotherapy for adolescents with depression: initial but no sustainable benefits. PMID- 19304906 TI - Surfactant protein C-deficient mice are susceptible to respiratory syncytial virus infection. AB - Patients with mutations in the pulmonary surfactant protein C (SP-C) gene develop interstitial lung disease and pulmonary exacerbations associated with viral infections including respiratory syncytial virus (RSV). Pulmonary infection with RSV caused more severe interstitial thickening, air space consolidation, and goblet cell hyperplasia in SP-C-deficient (Sftpc(-/-)) mice compared with SP-C replete mice. The RSV-induced pathology resolved more slowly in Sftpc(-/-) mice with lung inflammation persistent up to 30 days postinfection. Polymorphonuclear leukocyte and macrophage counts were increased in the bronchoalveolar lavage (BAL) fluid of Sftpc(-/-) mice. Viral titers and viral F and G protein mRNA were significantly increased in both Sftpc(-/-) and heterozygous Sftpc(+/-) mice compared with controls. Expression of Toll-like receptor 3 (TLR3) mRNA was increased in the lungs of Sftpc(-/-) mice relative to Sftpc(+/+) mice before and after RSV infection. Consistent with the increased TLR3 expression, BAL inflammatory cells were increased in the Sftpc(-/-) mice after exposure to a TLR3 specific ligand, poly(I:C). Preparations of purified SP-C and synthetic phospholipids blocked poly(I:C)-induced TLR3 signaling in vitro. SP-C deficiency increases the severity of RSV-induced pulmonary inflammation through regulation of TLR3 signaling. PMID- 19304907 TI - Lung environment determines unique phenotype of alveolar macrophages. AB - Alveolar macrophages (AM) are the most abundant antigen-presenting cells in the lungs, and they play a critical role in regulating pulmonary immune responses to inhaled pathogens and to allergens. However, compared with macrophages in other body sites, AM have an unusual phenotype that, in many respects, resembles the phenotype of dendritic cells (DC). Therefore, to more fully define the unique nature of AM, we compared the phenotype and function of AM with the phenotype and function of resident peritoneal lavage-derived macrophages (PLM). We found striking phenotypic differences between AM and PLM, particularly with regard to CD11c expression, and we also observed that AM had a significantly better antigen presenting capability than PLM. Therefore, we investigated the role of the local airway environment in generation of the unusual phenotype of AM. We carried out cell transfer experiments to compare macrophage differentiation in the airways with that in the peritoneal cavity. We observed significant upregulation of CD11c expression on bone marrow macrophages and peritoneal macrophages when they were adoptively transferred into the airways. In contrast, CD11c expression was not upregulated after cell transfer into the peritoneal cavity, whereas CD11b expression was significantly increased. In vitro, culture of bone marrow-adherent cells with surfactant protein D (SP-D) or granulocyte/macrophage colony stimulating factor (GM-CSF) induced significant upregulation of CD11c expression, and in vivo GM-CSF concentrations were significantly higher in bronchoalveolar than in peritoneal lavage fluid. Finally, GM-CSF(-/-) mice failed to develop CD11c(+) AM, but CD11c(+) AM were present in SP-D(-/-) mice. However, macrophages from GM-CSF(-/-) bone marrow could upregulate CD11c expression when transferred to the airways of wild-type mice. These results suggest that the airway environment promotes development of macrophages with unique DC-like characteristics and that this unusual phenotype is determined, to a large degree, by locally high concentrations of GM-CSF and, possibly, SP-D. PMID- 19304908 TI - Isolation of murine lung endothelial cells. AB - Several protocols for the isolation of endothelial cells (ECs) from murine lung have been described in the literature. We, however, encountered a number of problems while using these procedures that prevented us from consistently or reliably obtaining pure populations of ECs from the lungs of mice. By incorporating specific elements from previously published protocols, as well as adding some novel features, we developed a new strategy for isolating ECs from murine lung. In this approach, a suspension of lung cells is initially prepared from the lungs of 7- to 14-day-old mouse pups using procedures that prevent intravascular clotting and leukocyte activation, minimize mechanical trauma to the lung tissue, and limit exposure to the digesting enzymes. The resulting cell suspension is cultured for 2-3 days, trypsinized to produce a suspension of single cells, and then subjected to fluorescence-activated cell sorting using an anti-ICAM-2 antibody. The sorted cells are then plated and split 1:2 at each passage to maintain a high density of the cells. Using this approach, we have been able to isolate pure populations of ECs that were sustainable for extended periods in culture without the emergence of fibroblast overgrowth or the development of senescence. We believe the success of this approach will provide opportunities to take advantage of the large and growing number of knockout and transgenic mouse lines to investigate the endothelial-specific roles of targeted molecules in the pulmonary vasculature. PMID- 19304909 TI - Paracrine purinergic signaling determines lung endothelial nitric oxide production. AB - Although the vascular bed is a major source of nitric oxide (NO) production, factors regulating the production remain unclear. We considered the role played by paracrine signaling. Determinations by fluorescence microscopy in isolated, blood-perfused rat and mouse lungs revealed that a brief lung expansion enhanced cytosolic Ca(2+) (Ca(2+)cyt) oscillations in alveolar epithelial (AEC) and endothelial (EC) cells, and NO production in EC. Furthermore, as assessed by a novel microlavage assay, alveolar ATP production increased. Intra-alveolar microinfusion of the purinergic receptor antagonist, PPADS, and the nucleotide hydrolyzing enzyme, apyrase, each completely blocked the Ca(2+)cyt and NO responses in EC. Lung expansion induced Ca(2+)cyt oscillations in mice lacking the P2Y1, but not the P2Y2, purinergic receptors, which were located in the perivascular interstitium basolateral to AEC. Prolonged lung expansion instituted by mechanical ventilation at high tidal volume increased EC expression of nitrotyrosine, indicating development of nitrosative stress in lung microvessels. These findings reveal a novel mechanism in which mechanically induced purinergic signaling couples cross-compartmental Ca(2+)cyt oscillations to microvascular NO production. PMID- 19304910 TI - Mass spectrometric analysis of biomarkers and dilution markers in exhaled breath condensate reveals elevated purines in asthma and cystic fibrosis. AB - Exhaled breath condensate (EBC) analyses promise simple and noninvasive methods to measure airway biomarkers but pose considerable methodological challenges. We utilized mass spectrometry to measure EBC purine biomarkers adenosine and AMP plus urea to control for dilutional variability in two studies: 1) a cross sectional analysis of 28 healthy, 40 cystic fibrosis (CF), and 11 asthmatic children; and 2) a longitudinal analysis of 26 CF children before and after treatment of a pulmonary exacerbation. EBC adenosine, AMP, and urea were readily detected and quantified by mass spectrometry, and analysis suggested significant dilutional variability. Using biomarker-to-urea ratios to control for dilution, the EBC AMP-to-urea ratio was elevated in CF [median 1.3, interquartile range (IQR) 0.7-2.3] vs. control (median 0.75, IQR 0.3-1.4; P < 0.05), and the adenosine-to-urea ratio was elevated in asthma (median 1.5, IQR 0.9-2.9) vs. control (median 0.4, IQR 0.2-1.6; P < 0.05). Changes in EBC purine-to-urea ratios correlated with changes in percent predicted forced expiratory volume in 1 s (FEV(1)) (r = -0.53 AMP/urea, r = -0.55 adenosine/urea; P < 0.01 for both) after CF exacerbation treatment. Similar results were observed using dilution factors calculated from serum-to-EBC urea ratios or EBC electrolytes, and the comparable ratios of EBC electrolytes to urea in CF and control (median 3.2, IQR 1.6-6.0 CF; median 5.5, IQR 1.4-7.7 control) validated use of airway urea as an EBC dilution marker. These results show that mass spectrometric analyses can be applied to measurement of purines in EBC and demonstrate that EBC adenosine-to-urea and AMP to-urea ratios are potential noninvasive biomarkers of airways disease. PMID- 19304911 TI - Prolyl hydroxylase 2 deficiency limits proliferation of vascular smooth muscle cells by hypoxia-inducible factor-1{alpha}-dependent mechanisms. AB - Arterial O(2) levels are thought to modulate vascular smooth muscle cell (VSMC) proliferation and vascular remodeling, but the mechanisms involved are poorly understood. Here, we tested the hypothesis that PHD2, a prolyl hydroxylase domain (PHD)-containing O(2) sensor, modulates growth factor-induced proliferative responses of human pulmonary artery SMC (HPASMC). We found that both PHD1 and PHD2 were robustly expressed by HPASMC, and inhibiting prolyl hydroxylase activity pharmacologically by using the nonselective dioxygenase inhibitor dimethyloxalylglycine (DMOG) inhibited proliferation and cyclin A expression induced by PDGF-AB or FGF-2. Specific knockdown of PHD2 using small interfering RNAs had similar effects. The inhibitory effects of DMOG and PHD2 knockdown on proliferation and cyclin A expression were seen under both normoxic (20% O(2)) and moderately hypoxic (5% O(2)) conditions, and PHD2 expression was not affected by O(2) level nor by stimulation with PDGF or FGF-2, indicating that the proproliferative influence of PHD2 does not involve alterations of its expression. Knockdown of PHD2 increased hypoxia-inducible factor (HIF)-1alpha expression, as expected, but we also found that HIF-1alpha knockdown abolished the inhibitory effect of PHD2 knockdown on PDGF-induced cyclin A expression. Therefore, we conclude that PHD2 promotes growth factor-induced responses of human VSMC, acting by HIF-1alpha-dependent mechanisms. Given the role of PHD2 as an oxygen sensor in mammalian cells, these results raise the possibility that PHD2 links VSMC proliferation to O(2) availability. PMID- 19304912 TI - Hyperoxia-induced neonatal rat lung injury involves activation of TGF-{beta} and Wnt signaling and is protected by rosiglitazone. AB - Despite tremendous technological and therapeutic advances, bronchopulmonary dysplasia (BPD) remains a leading cause of respiratory morbidity in very low birth weight infants, and there are no effective preventive and/or therapeutic options. We have previously reported that hyperoxia-induced neonatal rat lung injury might be prevented by rosiglitazone (RGZ). Here, we characterize 1) perturbations in wingless/Int (Wnt) and transforming growth factor (TGF)-beta signaling, and 2) structural aberrations in lung morphology following 7-day continuous in vivo hyperoxia exposure to neonatal rats. We also tested whether treatment of neonatal pups with RGZ, concomitant to hyperoxia, could prevent such aberrations. Our study revealed that hyperoxia caused significant upregulation of Wnt signaling protein markers lymphoid enhancer factor 1 (Lef-1) and beta-catenin and TGF-beta pathway transducers phosphorylated Smad3 and Smad7 proteins in whole rat lung extracts. These changes were also accompanied by upregulation of myogenic marker proteins alpha-smooth muscle actin (alpha-SMA) and calponin but significant downregulation of the lipogenic marker peroxisome proliferator activated receptor-gamma (PPARgamma) expression. These molecular perturbations were associated with reduction in alveolar septal thickness, radial alveolar count, and larger alveoli in the hyperoxia-exposed lung. These hyperoxia-induced molecular and morphological changes were prevented by systemic administration of RGZ, with lung sections appearing near normal. This is the first evidence that in vivo hyperoxia induces activation of both Wnt and TGF-beta signal transduction pathways in lung and of its near complete prevention by RGZ. Hyperoxia-induced arrest in alveolar development, a hallmark of BPD, along with these molecular changes strongly implicates these proteins in hyperoxia-induced lung injury. Administration of PPARgamma agonists may thus be a potential strategy to attenuate hyperoxia-induced lung injury and subsequent BPD. PMID- 19304913 TI - PDE4 inhibitors roflumilast and rolipram augment PGE2 inhibition of TGF-{beta}1 stimulated fibroblasts. AB - Fibrotic diseases are characterized by the accumulation of extracellular matrix together with distortion and disruption of tissue architecture. Phosphodiesterase (PDE)4 inhibitors, by preventing the breakdown of cAMP, can inhibit fibroblast functions and may be able to mitigate tissue remodeling. Transforming growth factor (TGF)-beta1, a mediator of fibrosis, can potentially modulate cAMP by altering PGE(2) metabolism. The present study assessed whether PDE4 inhibitors functionally antagonize the profibrotic activity of fibroblasts stimulated by TGF beta1. The PDE4 inhibitors roflumilast and rolipram both inhibited fibroblast mediated contraction of three-dimensional collagen gels and fibroblast chemotaxis toward fibronectin in the widely studied human fetal lung fibroblast strain HFL-1 and several strains of fibroblasts from adult human lung. Roflumilast was approximately 10-fold more potent than rolipram. There was a trend for PDE4 inhibitors to inhibit more in the presence of TGF-beta1 (0.05 < P < 0.08). The effect of the PDE4 inhibitors was mediated through cAMP-stimulated protein kinase A (PKA), although a PKA-independent effect on gel contraction was also observed. The effect of PDE4 inhibitors depended on fibroblast production of PGE(2) and TGF beta1-induced PGE(2) production. PDE4 inhibitors together with TGF-beta1 resulted in augmented PGE(2) production together with increased expression of COX mRNA and protein. The present study supports the concept that PDE4 inhibitors may attenuate fibroblast activities that can lead to fibrosis and that PDE4 inhibitors may be particularly effective in the presence of TGF-beta1-induced fibroblast stimulation. PMID- 19304914 TI - Not what you expect: experience but not expectancy predicts conditioned responses in human visual and supplementary cortex. AB - When paired with aversive events, visual conditioned stimuli (CS) provoke increased activations in visual cortex. It is unclear however whether these changes reflect cognitive processes such as expectancy of the aversive unconditioned stimulus (US), or implicit associative learning of the contingencies outside awareness. Here, we used the "gambler's fallacy" phenomenon to parametrically and inversely manipulate the expectancy of an US and the number of conditioning trials: Increasing the number of CS-US pairings was associated with participants expecting the US to be less likely and vice versa. Magnetocortical activity evoked by the CS in occipital and supplementary motor areas was linearly related to the associative strength (number of CS-US pairings), but decreased as a function of expectancy. These results suggest that the cortical facilitation of fear cue processing is determined by associative strength and previous exposure to learning contingencies rather than by the cognitive anticipation for the US. PMID- 19304915 TI - Bladder cancer: diagnosis with diffusion-weighted MR imaging in patients with gross hematuria. AB - PURPOSE: To prospectively evaluate the usefulness of diffusion-weighted (DW) magnetic resonance (MR) imaging for the detection of bladder neoplasms in patients with gross hematuria of lower urinary tract origin. MATERIALS AND METHODS: The study protocol received institutional ethical committee approval, and informed consent was obtained. Between April 2007 and March 2008, 130 consecutive patients with gross hematuria whose upper urinary tract had a normal appearance at ultrasonographic examination were prospectively enrolled. Mean age was 59.4 years (range, 45-75 years). All patients were evaluated by using T2 weighted high-spatial-resolution MR imaging of the urinary bladder, followed by DW MR imaging. Two radiologists independently interpreted the T2-weighted and DW images, and discrepancies were resolved by consensus. Agreement was evaluated by using the kappa statistic. All patients underwent conventional cystoscopy. With cystoscopy and the final histopathologic findings as the reference standards, a comparison with imaging findings was performed by using the McNemar test. RESULTS: The consensus diagnostic performance of DW MR imaging for identification of bladder tumors was: sensitivity, 98.1% (104 of 106); specificity, 92.3% (24 of 26); PPV, 100% (104 of 104); negative predictive value, 92.3% (24 of 26); and accuracy, 97.0% (128 of 132). Two cases were falsely negative on T2-weighted MR images but were correctly diagnosed by using DW MR images. The agreement between DW MR imaging results and cystoscopic findings was excellent (kappa = 0.94) for identification of bladder neoplasm. DW MR imaging had a sensitivity and PPV of 98.5% (128 of 130) and 100% (128 of 128), respectively, for determining the cause of hematuria. CONCLUSION: DW MR imaging is a highly reliable imaging approach for identification of bladder tumors in patients with gross hematuria. PMID- 19304916 TI - Body tumor CT perfusion protocols: optimization of acquisition scan parameters in a rat tumor model. AB - PURPOSE: To evaluate the effects of total scanning time (TST), interscan delay (ISD), inclusion of image at peak vascular enhancement (IPVE), and selection of the input function vessel on the accuracy of tumor blood flow (BF) calculation with computed tomography (CT) in an animal model. MATERIALS AND METHODS: All animal protocols and experiments were approved by the institutional animal care and use committee prior to study initiation. After injection of 0.2 or 0.4 mL of iodinated contrast material, six rats with mammary adenocarcinoma (three tumors each) were scanned in the axial mode for 5 minutes with 1-second ISD (reference scan), 2.5-mm section thickness, 2.5-mm interval, pitch of 1.3, 120 kV, 240 mA, and 0.5-second rotation time. A total of 126 dynamic data sets were created with commercial software by varying TST and ISD, including or excluding the IPVE, and using the aorta or inferior vena cava (IVC) as the input function. Comparative analyses were used to test for significant differences (t test, Wilcoxon signed rank test). Regression analysis was performed to assess the relationship between attenuation of the input function vessel and BF. RESULTS: No significant difference was observed (P > .05) when TST was as short as 30 seconds (range, 20 23 mL/100 g). In sequences performed with an ISD longer than 8 seconds, BF was significantly elevated (P < .01). Inclusion of the IPVE eliminated this difference (P > .10). Use of the IVC as the input function resulted in significantly higher BF (P < .02), with a correlation between peak attenuation and BF (R(2) = 0.43). CONCLUSION: To reduce radiation dose in tumor perfusion with CT, TST can be reduced without causing significant changes in BF calculation in an animal model. Scanning the aortic reference with peak contrast enhancement reduces variability sufficiently to allow for longer ISDs. PMID- 19304917 TI - Chondroblastoma: radiofrequency ablation--alternative to surgical resection in selected cases. AB - PURPOSE: To demonstrate that radiofrequency (RF) ablation can be used safely and effectively to treat selected cases of chondroblastoma. MATERIALS AND METHODS: Approval was obtained from institutional review boards, research was in compliance with HIPAA protocol. The need to obtain informed consent was waived for retrospective review of patient records. The records of patients with biopsy proved chondroblastoma who were treated with RF ablation at two academic centers from July 1995 to July 2007 were reviewed. RF ablation was performed with a single-tip electrode by using computed tomography for guidance. Lesion characteristics were determined from imaging studies obtained at the time of the procedure. Symptoms were assessed before and 1 day after the procedure. Longer term follow-up was obtained from medical records. RESULTS: Thirteen male and four female patients were treated (mean age, 17.3 years). The lesions were located in the proximal humerus (n = 7), proximal tibia (n = 4), proximal femur (n = 3), and distal femur (n = 3). The mean volume of the lesions was 2.46 mL. All patients reported relief of symptoms on postprocedure day 1. Three patients were lost to follow-up. Of the 14 patients for whom longer-term (mean, 41.3 months; range, 4 134 months) follow-up was available, 12 had complete relief of symptoms with no need for medications and full return to all activities. The patient who had the largest lesion of the study required surgical intervention because of collapse of the articular surface in the treatment area. Residual viable tumor was found at surgery. Another patient experienced mechanical problems that were thought to be unrelated to the RF ablation and was rendered pain-free after subsequent surgical treatment. CONCLUSION: Percutaneous RF ablation is an alternative to surgery for treatment of selected chondroblastomas. Larger lesions beneath weight-bearing surfaces should be approached with caution due to an increased risk of articular collapse and recurrence. PMID- 19304918 TI - Small breast cancers: in vivo percutaneous US-guided radiofrequency ablation with dedicated cool-tip radiofrequency system. AB - PURPOSE: To evaluate in vivo the efficacy of a newly developed breast radiofrequency (RF) ablation system in human small invasive breast carcinomas in terms of induction of complete tumor necrosis, reproducibility of ablation lesion size and shape, and cosmetic outcome. MATERIALS AND METHODS: This study had institutional review board approval, and written informed consent was obtained. Thirty-four postmenopausal women (mean age, 53 years +/- 5 [standard deviation]; range, 49-62 years) with small (< or = 2 cm) biopsy-proved invasive ductal breast carcinomas were enrolled. RF energy was delivered through a 25-mm 15-gauge monopolar cool-tip needle electrode by using the temperature-controlled mode. Patients were divided into three groups according to their breast pattern as assessed at mammography. The volumetric size and geometry of the coagulation zone, together with ablation time, were determined. Histopathologic data were compared with postprocedural 3.0-T contrast material-enhanced magnetic resonance (MR) images. Cosmesis after RF ablation was assessed. Four weeks after RF ablation, patients underwent definitive surgery. RESULTS: All ablation procedures were performed successfully. For 97% of the procedures, nicotinamide adenine dinucleotide in its reduced form-diaphorase staining showed no evidence of viable cells. The mean induced ablation volume, as assessed with histologic analysis, was 12.50 cm(3) +/- 0.8. Tumor ablation volume on the postablation MR images showed good correlation with results of histopathologic analysis (r = 0.823, P < .005). No differences were observed in terms of duration of the procedure or ablation volume with respect to the glandular pattern of the breast (P > .05 for both). The general shape of the induced necrosis was close to a sphere in all cases. Cosmesis was excellent in 28 patients. CONCLUSION: A dedicated breast cool tip RF ablation system can induce complete tumor necrosis and reproducible ablation volumes independently of breast glandular pattern, providing excellent cosmesis. Postablation MR images are a reliable tool in predicting histologic findings. PMID- 19304919 TI - Pediatric 99mTc-DMSA SPECT performed by using iterative reconstruction with isotropic resolution recovery: improved image quality and reduced radiopharmaceutical activity. AB - PURPOSE: To compare two methods of reconstructing technetium 99m (99mTc) dimercaptosuccinic acid (DMSA) renal single photon emission computed tomographic (SPECT) data--ordered subset expectation maximization with three-dimensional resolution recovery (OSEM-3D) and filtered back projection (FBP)--in children in terms of improving image quality and reducing the radiopharmaceutical activity and radiation dose. MATERIALS AND METHODS: The institutional review board approved this retrospective study and waived the requirement for informed patient consent. Fifty sequential pediatric patient 99mTc-DMSA SPECT studies of 98 kidneys were retrospectively analyzed by using a dual-detector gamma camera. FBP reconstruction with data from both detectors was compared with OSEM-3D reconstruction with half the gamma photon counts (ie, data from only one detector). Two nuclear medicine physicians blinded to the patients' medical histories and reconstruction techniques evaluated the studies. Scores for image quality, renal size, and relative function were compared by using paired t tests. Total scores for renal cortical defects were compared by using the Wilcoxon signed rank test. The kappa coefficient was calculated as an indicator of the concordance between the OSEM-3D and FBP reconstruction methods. RESULTS: Image quality was significantly enhanced with OSEM-3D (P < .001, paired t test). Cortical defects were identified better on OSEM-3D images than on FBP images. Of the 98 kidney SPECT studies analyzed, 19 showed identical cortical defects and 75 showed none at both OSEM-3D and FBP. In four kidneys, OSEM-3D depicted cortical defects that were not seen with FBP. No significant difference in relative renal function between the two methods was observed (P = .973). CONCLUSION: Compared with FBP, OSEM-3D yielded superior image quality in the evaluation of 99mTc-DMSA renal SPECT data, with the potential for markedly reduced radiation doses and/or shorter scanning times for patients. PMID- 19304920 TI - Correlation of carotid plaque neovascularization detected by using contrast enhanced US with clinical symptoms. AB - PURPOSE: To determine the correlation between the degree of plaque enhancement with contrast agent microbubbles and clinical symptoms in patients with carotid atherosclerotic plaque. MATERIALS AND METHODS: The study was approved by the hospital ethical committee, and informed consent was obtained from all patients. One hundred four patients (83 men: mean age, 64 years +/- 9 [standard deviation]; 21 women: mean age, 61 years +/- 10) with carotid plaques were studied with standard and contrast material-enhanced ultrasonography (US). Contrast enhancement in the plaque was evaluated with visual interpretation and quantitative analysis. RESULTS: Among the 104 patients, 35 (34%) had transient ischemic attack and/or cerebrovascular ischemic stroke. Plaque enhancement was found in 28 (80%) of 35 symptomatic patients and in 21 (30%) of 69 asymptomatic patients (P < .001). Enhanced intensity in the plaque (13.9 dB +/- 6.4) and the ratio of enhanced intensity in the plaque to that in the lumen of the carotid artery (0.54 +/- 0.23) in symptomatic patients were significantly greater than those in asymptomatic patients (8.8 dB +/- 5.2 [P < .001] and 0.33 +/- 0.19 [P < .001], respectively). Sensitivity and specificity were 74% and 62%, respectively, for enhanced intensity in the plaque (cutoff value, 10.0 dB) and 74% and 75%, respectively, for ratio of enhanced intensity in the plaque to that in the lumen of the carotid artery (cutoff value, 0.46). CONCLUSION: Symptomatic patients had more intense contrast agent enhancement in the plaque than asymptomatic patients, suggesting that contrast-enhanced carotid US may be used for plaque risk stratification. PMID- 19304921 TI - Malignant liver tumors: treatment with percutaneous microwave ablation- complications among cohort of 1136 patients. AB - PURPOSE: To report the complications for percutaneous microwave (MW) ablation for the treatment of malignant liver tumors and the possible risk factors for complications in a large series of patients. MATERIALS AND METHODS: The study was approved by the institutional review board; informed consent was waived because of the retrospective design. Over a 13-year period, 1136 patients with 1928 malignant liver tumors underwent ultrasonographically guided percutaneous MW ablation (583 with a noncooled-shaft antenna and 553 with a cooled-shaft antenna). A total of 3697 MW ablation sessions (average, 1.8 sessions per patient) were performed. Mortality and treatment-related major and minor complications were documented. Data were subsequently analyzed to determine whether the major complication rate was related to antenna type, tumor size, tumor location, or number of MW sessions. RESULTS: Two deaths not directly attributable to MW ablation were encountered. Major complications occurred in 30 (2.6%) patients and included liver abscess and empyema (n = 5), bile duct injury (n = 2), perforation of the colon (n = 2), tumor seeding (n = 5), pleural effusion requiring thoracentesis (n = 12), hemorrhage requiring arterial embolization (n = 1), and skin burn requiring resection (n = 3). Minor complications included fever; pain; asymptomatic pleural effusion, gallbladder wall thickening, and arterioportal shunt; small stricture of the bile duct; and skin burn requiring no treatment. Use of noncooled-shaft antenna and an increased number of MW ablation sessions were associated with a higher rate of major complications (P < .05). CONCLUSION: MW ablation is a well-tolerated technique with an acceptably low rate of major complications for treatment of malignant liver tumors. Use of a cooled-shaft antenna, as well as fewer MW sessions, may help minimize major complications. PMID- 19304922 TI - Small-bowel neoplasms: prospective evaluation of MR enteroclysis. AB - PURPOSE: To prospectively evaluate the accuracy of magnetic resonance (MR) enteroclysis in the detection of small-bowel neoplasms in symptomatic patients, with conventional endoscopy, tissue specimen, capsule endoscopy, conventional enteroclysis, and follow-up findings as reference standards. MATERIALS AND METHODS: The study protocol was approved by the human research committee, and all patients gave written informed consent. One hundred fifty patients (83 male, 67 female; mean age, 42.6 years; age range, 17-84 years) who were clinically suspected of having small-bowel neoplasm and whose previous upper and lower gastrointestinal endoscopy findings were normal underwent MR enteroclysis. The MR enteroclysis findings were prospectively evaluated for the presence of focal bowel wall thickening, small-bowel masses, and small-bowel stenosis. Positive MR enteroclysis findings were compared with histopathologic examination results obtained after surgical (n = 19) or endoscopic (n = 2) procedures. Negative MR enteroclysis results were compared with the results of enteroscopy (n = 5), capsule endoscopy (n = 53), or conventional enteroclysis with subsequent clinical follow-up (n = 71). The diagnostic performance of MR enteroclysis was analyzed on a per-patient basis. RESULTS: MR enteroclysis was successfully completed in all 150 patients and enabled correct detection of 19 small-bowel neoplasms, which were confirmed at histopathologic examination: three carcinoid neoplasms, two adenocarcinomas, two stromal tumors, five lymphomas, one angiomatous mass, three small-bowel metastases, one leiomyoma, one adenoma, and one lipoma. Overall sensitivity, specificity, and accuracy in identifying patients with small-bowel lesions were 86% (19 of 22), 98% (126 of 128), and 97% (145 of 150), respectively. Two MR enteroclysis examinations yielded false-positive findings, and three yielded false-negative findings. CONCLUSION: MR enteroclysis is an accurate modality for detecting small-bowel neoplasms in symptomatic patients. PMID- 19304923 TI - Expanded hammerhead ribozymes containing addressable three-way junctions. AB - Recently, hammerhead ribozyme (HHR) motifs have been utilized as powerful tools for gene regulation. Here we present a novel design of expanded full-length HHRs that allows attaching additional functionalities to the ribozyme. These features allowed us to construct a very efficient artificial riboswitch in bacteria. Following the design of naturally occurring three-way junctions we attached an additional helix (IV) to stem I of the HHR while maintaining very fast cleavage rates. We found that the cleavage activity strongly depends on the exact design of the junction site. Incorporation of the novel ribozyme scaffold into a bacterial mRNA allowed the control of gene expression mediated by autocatalytic cleavage of the ribozyme. Appending an aptamer to the newly introduced stem enabled the identification of very powerful theophylline-inducible RNA switches by in vivo screening. Further investigations revealed a cascading system operating beyond the ribozyme-dependent mechanism. In conclusion, we extended the hammerhead toolbox for synthetic biology applications by providing an additional position for the attachment of regulatory modules for in vivo control of gene expression. PMID- 19304924 TI - Multiple independent domains of dGW182 function in miRNA-mediated repression in Drosophila. AB - miRNA-mediated repression affects a wide range of biological processes including development and human pathologies. The GW182 protein is a key component of miRNA repression complex, recruited by Argonaute and functioning downstream to repress translation and accelerate mRNA degradation, but little is known about how GW182 proteins act. Using both tethered function and complementation assays, we identify three independent domains of the Drosophila GW182 protein (also termed Gawky) that are sufficient to repress mRNA. Each of these domains also functions independently of poly(A) tails. These results indicate that miRNA-mediated repression is facilitated by multiple domains of GW182. PMID- 19304925 TI - Importance of the C-terminal domain of the human GW182 protein TNRC6C for translational repression. AB - Proteins of the GW182 family play an important role in the execution of microRNA repression in metazoa. They interact directly with Argonaute proteins, components of microRNPs, and also form part of P-bodies, structures implicated in translational repression and mRNA degradation. Recent results demonstrated that Drosophila GW182 has the potential to both repress translation and accelerate mRNA deadenylation and decay. In contrast to a single GW182 protein in Drosophila, the three GW182 paralogs TNRC6A, TNRC6B, and TNRC6C are encoded in mammalian genomes. In this study, we provide evidence that TNRC6C, like TNRC6A and TNRC6B, is important for efficient miRNA repression. We further demonstrate that tethering of each of the human TNRC6 proteins to a reporter mRNA has a dramatic inhibitory effect on protein synthesis. The repression is due to a combination of effects on the mRNA level and mRNA translation. Through deletion and mutagenesis, we identified the C-terminal part of TNRC6C encompassing the RRM RNA-binding motif as a key effector domain mediating protein synthesis repression by TNRC6C. PMID- 19304926 TI - Polyadenylation releases mRNA from RNA polymerase II in a process that is licensed by splicing. AB - When transcription is coupled to pre-mRNA processing in HeLa nuclear extracts nascent transcripts become attached to RNA polymerase II during assembly of the cleavage/polyadenylation apparatus (CPA), and are not released even after cleavage at the poly(A) site. Here we show that these cleaved transcripts are anchored to the polymerase at their 3' ends by the CPA or, when introns are present, by the larger 3'-terminal exon definition complex (EDC), which consists of splicing factors complexed with the CPA. Poly(A) addition releases the RNA from the polymerase when the RNA is anchored only by the CPA. When anchored by the EDC, poly(A) addition remains a requirement, but it triggers release only after being licensed by splicing. The process by which RNA must first be attached to the polymerase by the EDC, and then can only be released following dual inputs from splicing and polyadenylation, provides an obvious opportunity for surveillance as the RNA enters the transport pathway. PMID- 19304927 TI - Which patient-related factors determine self-perceived patient adherence to prescribed dyspepsia medication? AB - Patient adherence to medical treatment for dyspepsia is disappointing, whereas adherence is crucial for a proper evaluation of treatment. This prospective study used elements of the Integrated Change Model and Weiner's Attribution Theory to describe patients' important cognitions and their interrelationships regarding self-perceived adherence to short-term medical treatment for dyspepsia. Patient questionnaires measured the predictors before treatment and self-perceived adherence after treatment. Approximately one-quarter of the patients indicated that they were non-adherent (n = 347). Univariate and multiple linear regression analyses revealed several significant predictors that explained 44% of the variance in self-perceived adherence. Patients with a low educational level, patients who claimed to regularly forget their medication in general, patients with a low self-efficacy or a low intention were less likely to be adherent. These results may indicate targets for interventions designed to improve adherence to medical treatment for dyspepsia. For instance, asking about expected difficulties in taking acid suppressants (e.g. forgetfulness or medication use at weekends) and making action plans to overcome these difficulties (e.g. using reminders) may result in improved adherence rates. Such an approach may reach a substantial number of patients since one in five patients in our study experienced some difficulties in taking medication. PMID- 19304928 TI - Process evaluation of a school-based weight gain prevention program: the Dutch Obesity Intervention in Teenagers (DOiT). AB - Health promotion programs benefit from an accompanying process evaluation since it can provide more insight in the strengths and weaknesses of a program. A process evaluation was conducted to assess the reach, implementation, satisfaction and maintenance of a school-based program aimed at the prevention of excessive weight gain among Dutch adolescents [Dutch Obesity Intervention in Teenagers (DOiT)]. Our process evaluation involved data collections by means of questionnaires among students, teachers, school board and site staff. The results indicated immense difficulties in the recruitment phase and therefore a low reach at school level. However, among adolescents of the schools that participated, the reach was high (84%). Furthermore, the classroom intervention was implemented successfully based on the number of lessons taught. Most teachers rated the DOiT intervention positively; students rated the intervention 6.6 on a scale of 1-10. The majority of the teachers planned to implement the DOiT-intervention program in the future, as they perceived DOiT feasible for pre-vocational education students. PMID- 19304929 TI - The analysis of Arabidopsis nicotianamine synthase mutants reveals functions for nicotianamine in seed iron loading and iron deficiency responses. AB - Nicotianamine chelates and transports micronutrient metal ions in plants. It has been speculated that nicotianamine is involved in seed loading with micronutrients. A tomato (Solanum lycopersicum) mutant (chloronerva) and a tobacco (Nicotiana tabacum) transgenic line have been utilized to analyze the effects of nicotianamine loss. These mutants showed early leaf chlorosis and had sterile flowers. Arabidopsis (Arabidopsis thaliana) has four NICOTIANAMINE SYNTHASE (NAS) genes. We constructed two quadruple nas mutants: one had full loss of NAS function, was sterile, and showed a chloronerva-like phenotype (nas4x-2); another mutant, with intermediate phenotype (nas4x-1), developed chlorotic leaves, which became severe upon transition from the vegetative to the reproductive phase and upon iron (Fe) deficiency. Residual nicotianamine levels were sufficient to sustain the life cycle. Therefore, the nas4x-1 mutant enabled us to study late nicotianamine functions. This mutant had no detectable nicotianamine in rosette leaves of the reproductive stage but low nicotianamine levels in vegetative rosette leaves and seeds. Fe accumulated in the rosette leaves, while less Fe was present in flowers and seeds. Leaves, roots, and flowers showed symptoms of Fe deficiency, whereas leaves also showed signs of sufficient Fe supply, as revealed by molecular-physiological analysis. The mutant was not able to fully mobilize Fe to sustain Fe supply of flowers and seeds in the normal way. Thus, nicotianamine is needed for correct supply of seeds with Fe. These results are fundamental for plant manipulation approaches to modify Fe homeostasis regulation through alterations of NAS genes. PMID- 19304930 TI - The synthetic elicitor 3,5-dichloroanthranilic acid induces NPR1-dependent and NPR1-independent mechanisms of disease resistance in Arabidopsis. AB - Immune responses of Arabidopsis (Arabidopsis thaliana) are at least partially mediated by coordinated transcriptional up-regulation of plant defense genes, such as the Late/sustained Up-regulation in Response to Hyaloperonospora parasitica (LURP) cluster. We found a defined region in the promoter of the LURP member CaBP22 to be important for this response. Using a CaBP22 promoter-reporter fusion, we have established a robust and specific high-throughput screening system for synthetic defense elicitors that can be used to trigger defined subsets of plant immune responses. Screening a collection of 42,000 diversity oriented molecules, we identified 114 candidate LURP inducers. One representative, 3,5-dichloroanthranilic acid (DCA), efficiently induced defense reactions to the phytopathogens H. parasitica and Pseudomonas syringae. In contrast to known salicylic acid analogs, such as 2,6-dichloroisonicotinic acid (INA), which exhibit a long-lasting defense-inducing activity and are fully dependent on the transcriptional cofactor NPR1 (for Nonexpresser of Pathogenesis Related genes1), DCA acts transiently and is only partially dependent on NPR1. Microarray analyses revealed a cluster of 142 DCA- and INA-responsive genes that show a pattern of differential expression coinciding with the kinetics of DCA mediated disease resistance. These ACID genes (for Associated with Chemically Induced Defense) constitute a core gene set associated with chemically induced disease resistance, many of which appear to encode components of the natural immune system of Arabidopsis. PMID- 19304931 TI - Phospholipase D activation is an early component of the salicylic acid signaling pathway in Arabidopsis cell suspensions. AB - Salicylic acid (SA) plays a central role in defense against pathogen attack, as well as in germination, flowering, senescence, and the acquisition of thermotolerance. In this report we investigate the involvement of phospholipase D (PLD) in the SA signaling pathway. In presence of exogenous primary alcohols, the production of phosphatidic acid by PLD is diverted toward the formation of phosphatidylalcohols through a reaction called transphosphatidylation. By in vivo metabolic phospholipid labeling with (33)P(i), PLD activity was found to be induced 45 min after addition of SA. We show that incubation of Arabidopsis (Arabidopsis thaliana) cell suspensions with primary alcohols inhibited the induction of two SA-responsive genes, PATHOGENESIS-RELATED1 and WRKY38, in a dose dependent manner. This inhibitory effect was more pronounced when the primary alcohols were more hydrophobic. Secondary or tertiary alcohols had no inhibitory effect. These results provide compelling arguments for PLD activity being upstream of the induction of these genes by SA. A subsequent study of n-butanol effects on the SA-responsive transcriptome identified 1,327 genes differentially expressed upon SA treatment. Strikingly, the SA response of 380 of these genes was inhibited by n-butanol but not by tert-butanol. A detailed analysis of the regulation of these genes showed that PLD could act both positively and negatively, either on gene induction or gene repression. The overlap with the previously described phosphatidylinositol-4-kinase pathway is discussed. PMID- 19304932 TI - A new chlorophyll degradation pathway. PMID- 19304933 TI - Disruption of adenosine-5'-phosphosulfate kinase in Arabidopsis reduces levels of sulfated secondary metabolites. AB - Plants can metabolize sulfate by two pathways, which branch at the level of adenosine 5'-phosphosulfate (APS). APS can be reduced to sulfide and incorporated into Cys in the primary sulfate assimilation pathway or phosphorylated by APS kinase to 3'-phosphoadenosine 5'-phosphosulfate, which is the activated sulfate form for sulfation reactions. To assess to what extent APS kinase regulates accumulation of sulfated compounds, we analyzed the corresponding gene family in Arabidopsis thaliana. Analysis of T-DNA insertion knockout lines for each of the four isoforms did not reveal any phenotypical alterations. However, when all six combinations of double mutants were compared, the apk1 apk2 plants were significantly smaller than wild-type plants. The levels of glucosinolates, a major class of sulfated secondary metabolites, and the sulfated 12 hydroxyjasmonate were reduced approximately fivefold in apk1 apk2 plants. Although auxin levels were increased in the apk1 apk2 mutants, as is the case for most plants with compromised glucosinolate synthesis, typical high auxin phenotypes were not observed. The reduction in glucosinolates resulted in increased transcript levels for genes involved in glucosinolate biosynthesis and accumulation of desulfated precursors. It also led to great alterations in sulfur metabolism: the levels of sulfate and thiols increased in the apk1 apk2 plants. The data indicate that the APK1 and APK2 isoforms of APS kinase play a major role in the synthesis of secondary sulfated metabolites and are required for normal growth rates. PMID- 19304934 TI - The ESCRT-related CHMP1A and B proteins mediate multivesicular body sorting of auxin carriers in Arabidopsis and are required for plant development. AB - Plasma membrane proteins internalized by endocytosis and targeted for degradation are sorted into lumenal vesicles of multivesicular bodies (MVBs) by the endosomal sorting complexes required for transport (ESCRT) machinery. Here, we show that the Arabidopsis thaliana ESCRT-related CHARGED MULTIVESICULAR BODY PROTEIN/CHROMATIN MODIFYING PROTEIN1A (CHMP1A) and CHMP1B proteins are essential for embryo and seedling development. Double homozygous chmp1a chmp1b mutant embryos showed limited polar differentiation and failed to establish bilateral symmetry. Mutant seedlings show disorganized apical meristems and rudimentary true leaves with clustered stomata and abnormal vein patterns. Mutant embryos failed to establish normal auxin gradients. Three proteins involved in auxin transport, PINFORMED1 (PIN1), PIN2, and AUXIN-RESISTANT1 (AUX1) mislocalized to the vacuolar membrane of the mutant. PIN1 was detected in MVB lumenal vesicles of control cells but remained in the limiting membrane of chmp1a chmp1b MVBs. The chmp1a chmp1b mutant forms significantly fewer MVB lumenal vesicles than the wild type. Furthermore, CHMP1A interacts in vitro with the ESCRT-related proteins At SKD1 and At LIP5. Thus, Arabidopsis CHMP1A and B are ESCRT-related proteins with conserved endosomal functions, and the auxin carriers PIN1, PIN2, and AUX1 are ESCRT cargo proteins in the MVB sorting pathway. PMID- 19304935 TI - Characterization of Solanum tuberosum multicystatin and its structural comparison with other cystatins. AB - Potato (Solanum tuberosum) multicystatin (PMC) is a crystalline Cys protease inhibitor present in the subphellogen layer of potato tubers. It consists of eight tandem domains of similar size and sequence. Our in vitro results showed that the pH/PO(4)(-)-dependent oligomeric behavior of PMC was due to its multidomain nature and was not a characteristic of the individual domains. Using a single domain of PMC, which still maintains inhibitor activity, we identified a target protein of PMC, a putative Cys protease. In addition, our crystal structure of a representative repeating unit of PMC, PMC-2, showed structural similarity to both type I and type II cystatins. The N-terminal trunk, alpha helix, and L2 region of PMC-2 were most similar to those of type I cystatins, while the conformation of L1 more closely resembled that of type II cystatins. The structure of PMC-2 was most similar to the intensely sweet protein monellin from Dioscorephyllum cumminisii (serendipity berry), despite a low level of sequence similarity. We present a model for the possible molecular organization of the eight inhibitory domains in crystalline PMC. The unique molecular properties of the oligomeric PMC crystal are discussed in relation to its potential function in regulating the activity of proteases in potato tubers. PMID- 19304936 TI - Pheophytin pheophorbide hydrolase (pheophytinase) is involved in chlorophyll breakdown during leaf senescence in Arabidopsis. AB - During leaf senescence, chlorophyll is removed from thylakoid membranes and converted in a multistep pathway to colorless breakdown products that are stored in vacuoles. Dephytylation, an early step of this pathway, increases water solubility of the breakdown products. It is widely accepted that chlorophyll is converted into pheophorbide via chlorophyllide. However, chlorophyllase, which converts chlorophyll to chlorophyllide, was found not to be essential for dephytylation in Arabidopsis thaliana. Here, we identify pheophytinase (PPH), a chloroplast-located and senescence-induced hydrolase widely distributed in algae and land plants. In vitro, Arabidopsis PPH specifically dephytylates the Mg-free chlorophyll pigment, pheophytin (phein), yielding pheophorbide. An Arabidopsis mutant deficient in PPH (pph-1) is unable to degrade chlorophyll during senescence and therefore exhibits a stay-green phenotype. Furthermore, pph-1 accumulates phein during senescence. Therefore, PPH is an important component of the chlorophyll breakdown machinery of senescent leaves, and we propose that the sequence of early chlorophyll catabolic reactions be revised. Removal of Mg most likely precedes dephytylation, resulting in the following order of early breakdown intermediates: chlorophyll --> pheophytin --> pheophorbide. Chlorophyllide, the last precursor of chlorophyll biosynthesis, is most likely not an intermediate of breakdown. Thus, chlorophyll anabolic and catabolic reactions are metabolically separated. PMID- 19304937 TI - A single vegetative actin isovariant overexpressed under the control of multiple regulatory sequences is sufficient for normal Arabidopsis development. AB - The relative significance of gene regulation and protein isovariant differences remains unexplored for most gene families, particularly those participating in multicellular development. Arabidopsis thaliana encodes three vegetative actins, ACT2, ACT7, and ACT8, in two ancient and highly divergent subclasses. Mutations in any of these differentially expressed actins revealed only mild phenotypes. However, double mutants were extremely dwarfed, with altered cell and organ morphology and an aberrant F-actin cytoskeleton (e.g., act2-1 act7-4 and act8-2 act7-4) or totally root-hairless (e.g., act2-1 act8-2). Our studies suggest that the three vegetative actin genes and protein isovariants play distinct subclass specific roles during plant morphogenesis. For example, during root development, ACT7 was involved in root growth, epidermal cell specification, cell division, and root architecture, and ACT2 and ACT8 were essential for root hair tip growth. Also, genetic complementation revealed that the ACT2 and ACT8 isovariants, but not ACT7, fully rescued the root hair growth defects of single and double mutants. Moreover, we synthesized fully normal plants overexpressing the ACT8 isovariant from multiple actin regulatory sequences as the only vegetative actin in the act2-1 act7-4 background. In summary, it is evident that differences in vegetative actin gene regulation and the diversity in actin isovariant sequences are essential for normal plant development. PMID- 19304938 TI - SHALLOT-LIKE1 is a KANADI transcription factor that modulates rice leaf rolling by regulating leaf abaxial cell development. AB - As an important agronomic trait, rice (Oryza sativa L.) leaf rolling has attracted much attention from plant biologists and breeders. Moderate leaf rolling increases the photosynthesis of cultivars and hence raises grain yield. However, the relevant molecular mechanism remains unclear. Here, we show the isolation and functional characterization of SHALLOT-LIKE1 (SLL1), a key gene controlling rice leaf rolling. sll1 mutant plants have extremely incurved leaves due to the defective development of sclerenchymatous cells on the abaxial side. Defective development can be functionally rescued by expression of SLL1. SLL1 is transcribed in various tissues and accumulates in the abaxial epidermis throughout leaf development. SLL1 encodes a SHAQKYF class MYB family transcription factor belonging to the KANADI family. SLL1 deficiency leads to defective programmed cell death of abaxial mesophyll cells and suppresses the development of abaxial features. By contrast, enhanced SLL1 expression stimulates phloem development on the abaxial side and suppresses bulliform cell and sclerenchyma development on the adaxial side. Additionally, SLL1 deficiency results in increased chlorophyll and photosynthesis. Our findings identify the role of SLL1 in the modulation of leaf abaxial cell development and in sustaining abaxial characteristics during leaf development. These results should facilitate attempts to use molecular breeding to increase the photosynthetic capacity of rice, as well as other crops, by modulating leaf development and rolling. PMID- 19304940 TI - Insulin sensitivity as a key mediator of growth hormone actions on longevity. AB - Reduced insulin sensitivity and glucose intolerance have been long suspected of having important involvement in aging. Here we report that in studies of calorie restriction (CR) effects in mutant (Prop1(df) and growth hormone receptor knockout [GHRKO]) and normal mice, insulin sensitivity was strongly associated with longevity. Of particular interest was enhancement of the already increased insulin sensitivity in CR df/df mice in which longevity was also further extended and the lack of changes in insulin sensitivity in calorically restricted GHRKO mice in which there was no further increase in average life span. We suggest that enhanced insulin sensitivity, in conjunction with reduced insulin levels, may represent an important (although almost certainly not exclusive) mechanism of increased longevity in hypopituitary, growth hormone (GH)-resistant, and calorie restricted animals. We also report that the effects of GH treatment on insulin sensitivity may be limited to the period of GH administration. PMID- 19304939 TI - Exercise training modulates the nitric oxide synthase profile in skeletal muscle from old rats. AB - The effects of exercise training on the nitric oxide synthase (NOS) isoform profile in aging fast-twitch (white gastrocnemius [WG]) and slow-twitch (soleus [SOL]) muscle have not been investigated. Six-month and 27-month male Fischer-344 rats were divided into the following groups: young sedentary (YS), young treadmill exercise trained for 12 weeks, old sedentary (OS), and old exercise trained (OE). Inducible NOS (iNOS) protein expression and activity were significantly higher in OS compared with YS, whereas exercise training significantly reduced iNOS protein and activity levels in the WG. Neuronal NOS protein expression decreased with aging in WG but was upregulated significantly with exercise training in OE for both WG and SOL. Endothelial NOS (eNOS) protein levels were depressed in WG of old rats but were higher in OE than in OS. eNOS was unaffected by aging or exercise in the SOL. Our results indicate that endurance exercise training attenuates age-induced alterations of NOS isoforms with a greater response in fast-twitch compared with slow-twitch muscle. PMID- 19304941 TI - Acute stroke intervention. PMID- 19304942 TI - Apelin prevents aortic aneurysm formation by inhibiting macrophage inflammation. AB - Apelin is a potent inodilator with recently described antiatherogenic properties. We hypothesized that apelin might also attenuate abdominal aortic aneurysm (AAA) formation by limiting disease-related vascular wall inflammation. C57BL/6 mice implanted with osmotic pumps filled with apelin or saline were treated with pancreatic elastase to create infrarenal AAAs. Mice were euthanized for aortic PCR analysis or followed ultrasonographically and then euthanized for histological analysis. The cellular expression of inflammatory cytokines and chemokines in response to apelin was also assessed in cultured macrophages, smooth muscle cells, and fibroblasts. Apelin treatment resulted in diminished AAA formation, with a 47% reduction in maximal cross-sectional area (0.74 vs. 1.39 mm(2), P < 0.03) and a 57% reduction in macrophage infiltrate (113 vs. 261.3 cells/high-power field, P < 0.0001) relative to the saline-treated group. Apelin infusion was also associated with significantly reduced aortic macrophage colony stimulating factor expression and decreased monocyte chemattractant protein (MCP) 1, macrophage inflammatory protein (MIP)-1alpha, interleukin (IL)-6, and tumor necrosis factor (TNF)-alpha mean mRNA levels. Apelin stimulation of cultured macrophages significantly reduced MCP-1 and TNF-alpha mRNA levels relative to baseline (2.03- and 1.89-fold reduction, P < 0.03, respectively) but did not affect intimal adhesion molecule expression or medial or adventitial cell cytokine production. Apelin significantly reduces aneurysm formation in the elastase model of human AAA disease. The mechanism appears to be decreased macrophage burden, perhaps related to an apelin-mediated decrease in proinflammatory cytokine and chemokine activation. PMID- 19304943 TI - Activation of NF-kappaB is a critical element in the antiapoptotic effect of anesthetic preconditioning. AB - Anesthetic preconditioning (APC), defined as brief exposure to inhalational anesthetics before cardiac ischemia-reperfusion (I/R), limits injury in both animal models and in humans. APC can result in the production of reactive oxygen species (ROS), and prior work has shown that APC can modify activation of NF kappaB during I/R, with consequent reduction in the expression of inflammatory mediators. However, the role of NF-kappaB activation before I/R is unknown. Therefore, these experiments tested the hypothesis that APC-induced ROS results in activation of NF-kappaB before I/R, with consequent increased expression of antiapoptotic proteins such as Bcl-2 and decreased apoptosis. Experiments utilized an established perfused heart rat model of sevoflurane APC and I/R. The role of NF-kappaB was defined by a novel method of transient inhibition of the regulatory kinase IKK using the reversible inhibitor SC-514. In addition to functional measures of left ventricular developed and end-diastolic pressure, phosphorylation of IkappaBalpha and activation of NF-kappaB were measured along with cytosolic protein content of Bcl-2, release of cytochrome c, and degradation of caspase-3. APC resulted in ROS-dependent phosphorylation of IkappaBalpha and activation of NF-kappaB before I/R. APC also increased the expression of Bcl-2 before I/R. In addition to functional protection following I/R, APC resulted in lower release of cytochrome c and caspase-3 degradation. These protective effects of APC were abolished by transient inhibition of IkappaBalpha phosphorylation and NF-kappaB activation by SC-514 followed by washout. ROS-dependent activation of NF-kappaB by APC before I/R is a critical element in the protective effect of APC. APC reduces apoptosis and functional impairment by increasing Bcl-2 expression before I/R. Interventions that increase NF-kappaB activation before I/R should protect hearts from I/R injury. PMID- 19304944 TI - Thrombospondin-1 modulates VEGF-A-mediated Akt signaling and capillary survival in the developing retina. AB - Microvascular development is often perceived to result from a balance of positive and negative factors that impact signaling for proliferation and survival. The survival signaling that results from hypoxia-induced VEGF-A has been well established, but the factors that antagonize this signaling have been poorly studied. As endogenous inhibitors of angiogenesis, thrombospondins (TSPs) are likely candidates to affect survival signaling. Here we report that TSP1 antagonized microvascular survival to retinal hyperoxia, and Akt signaling in both the retina and in cultured endothelial cells. TSP1 expression is correlated with the association of the CD36 receptor with Src versus Fyn. In the presence of TSP1, CD36 is coprecipitated with Fyn as previously shown by others. However, in the absence of TSP1, there is a preferential association with Src. We now demonstrate that these Src family kinases play an important role in modulating microvascular survival in response to TSP1 by crossing tsp1(-/-) mice to the src( /-) and fyn(-/-) mice and testing the survival of retinal blood vessels in hyperoxia. We find that tsp1(-/-), fyn(-/-), and double-mutant tsp1(-/-)/fyn(-/-) mice have a similar enhancement of capillary survival in oxygen, whereas in a tsp(-/-) background, the loss of only one allele of src restores the balance in survival and apoptosis to that of wild-type mice. Taken together, we hypothesize that TSP1 antagonizes VEGF-driven Akt survival signaling in part through the recruitment of Fyn to membrane domains containing CD36, but when TSP1 is absent, an opposing Src recruitment contributes to VEGF-driven Akt phosphorylation and capillary survival. PMID- 19304945 TI - Microarray analysis of gene expression in mouse aorta reveals role of the calcium signaling pathway in control of atherosclerosis susceptibility. AB - Inbred mouse strains C57BL/6J (B6) and C3H/HeJ (C3H) exhibit a marked difference in atherosclerotic lesion formation when deficient in apolipoprotein E (apoE(-/ )), and the arterial wall has been identified as a source of the difference in atherosclerosis susceptibility. In the present study, differences in gene expression in aortic walls of the two strains were analyzed by microarrays. Total RNA was extracted from the aorta of 6-wk-old female B6 and C3H apoE(-/-) mice fed a chow or Western diet. There were 1,514 genes in chow fed mice and 590 genes in Western fed mice that were found to be differentially expressed between the two strains. Pathway analysis of differentially expressed genes suggested a role for the calcium signaling pathway in regulating atherosclerosis susceptibility. Oxidized LDL (oxLDL) induced a dose-dependent rise in cytosolic calcium levels in B6 endothelial cells. oxLDL-induced monocyte chemoattractant protein-1 production was inhibited by pretreatment with calcium chelator EGTA or intracellular calcium trapping compound BAPTA, indicating that calcium ions mediate the effect of oxLDL on monocyte chemoattractant protein-1 induction. The present findings demonstrate involvement of the calcium signaling pathway in the inflammatory process of atherogenesis. PMID- 19304946 TI - Epoxyeicosanoids as mediators of neurogenic vasodilation in cerebral vessels. AB - Epoxyeicosatrienoic acids (EETs) are potent vasodilators produced from arachidonic acid by cytochrome P-450 (CYP) epoxygenases and metabolized to vicinal diols by soluble epoxide hydrolase (sEH). In the brain, EETs are produced by astrocytes and the vascular endothelium and are involved in the control of cerebral blood flow (CBF). Recent evidence, however, suggests that epoxygenases and sEH are present in perivascular vasodilator nerve fibers innervating the cerebral surface vasculature. In the present study, we tested the hypothesis that EETs are nerve-derived relaxing factors in the cerebral circulation. We first traced these fibers by retrograde labeling in the rat to trigeminal ganglia (TG) and sphenopalatine ganglia (SPG). We then examined the expression of CYP epoxygenases and sEH in these ganglia. RT-PCR and Western blot analysis identified CYP2J3 and CYP2J4 epoxygenase isoforms and sEH in both TG and SPG, and immunofluorescence double labeling identified CYP2J and sEH immunoreactivity in neuronal cell bodies of both ganglia. To evaluate the functional role of EETs in neurogenic vasodilation, we elicited cortical hyperemia by electrically stimulating efferent cerebral perivascular nerve fibers and by chemically stimulating oral trigeminal fibers with capsaicin. Cortical blood flow responses were monitored by laser-Doppler flowmetry. Local administration to the cortical surface of the putative EET antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (30 mumol/l) attenuated CBF responses to electrical and chemical stimulation. These results suggest that EETs are produced by perivascular nerves and play a role in neurogenic vasodilation of the cerebral vasculature. The findings have important implications to such clinical conditions as migraine, vasospasm after subarachnoid hemorrhage, and stroke. PMID- 19304947 TI - Hypoxia during pregnancy in rats leads to early morphological changes of atherosclerosis in adult offspring. AB - Exposure to an adverse intrauterine environment increases the risk of cardiovascular disease later in adult life. However, the time course relationship between prenatal hypoxia and the onset of atherosclerosis in offspring remains unknown. The purpose of this study is to evaluate the role of reduced fetal oxygen supply on early development of atherogenesis in the adult offspring and further assess its susceptibility to sex-, hyperlipidemia-, and postnatal hypoxemia-related differences. Based on a 4 x 2 full factorial design consisting of four factors of maternal hypoxia, sex, hyperlipidemia, and postnatal hypoxemia, characteristics of growth were determined, and histopathological observation and morphometric analysis of the thoracic aortas were performed in Sprague-Dawley rat offspring. Intrauterine growth restriction, altered body shape at birth, and accelerated postnatal weight gain occurred in the maternal hypoxia group but did not occur in the control group. In 16-mo-old maternal hypoxia offspring, the thoracic aortas exhibited lesions similar to early events in atherosclerosis that involved impaired endothelial cells, thickening and fibration of intimas, infiltration of inflammatory cells to the subendothelial space, and migration and proliferation of vascular smooth muscle cells to the intima. In contrast, no detectable pathological changes were observed in the offspring without maternal hypoxia exposure. Morphometric analysis further demonstrated that prenatal hypoxia caused a significant thickening of intima (P < 0.001) with a main effect of 5.5 mum, an approximately twofold increase compared with controls. In addition, there was a positive additive relationship between prenatal hypoxia and hyperlipidemia on the intimal thickness (P < 0.05). There were no other main effects or interaction among these four factors. In summary, our results indicate that maternal hypoxia during pregnancy leads to early pathological appearances of atherogenesis in adult offspring. This effect was enhanced with hyperlipemia but was unaffected by postnatal hypoxia or sex. PMID- 19304948 TI - Cardiovascular prevention by dietary nitrate and nitrite. PMID- 19304949 TI - Targeted ablation of cardiac sympathetic neurons reduces resting, reflex and exercise-induced sympathetic activation in conscious rats. AB - Cholera toxin B subunit conjugated to saporin (SAP, a ribosomal inactivating protein that binds to and inactivates ribosomes) was injected in both stellate ganglia to evaluate the physiological response to targeted ablation of cardiac sympathetic neurons. Resting cardiac sympathetic activity (cardiac sympathetic tonus), exercise-induced sympathetic activity (heart rate responses to graded exercise), and reflex sympathetic activity (heart rate responses to graded doses of sodium nitroprusside, SNP) were determined in 18 adult conscious Sprague Dawley male rats. Rats were randomly divided into the following three groups (n = 6/group): 1) control (no injection), 2) bilateral stellate ganglia injection of unconjugated cholera toxin B (CTB), and 3) bilateral stellate ganglia injection of cholera toxin B conjugated to SAP (CTB-SAP). CTB-SAP rats, compared with control and CTB rats, had reduced cardiac sympathetic tonus and reduced heart rate responses to graded exercise and graded doses of SNP. Furthermore, the number of stained neurons in the stellate ganglia and spinal cord (segments T(1) T(4)) was reduced in CTB-SAP rats. Thus CTB-SAP retrogradely transported from the stellate ganglia is effective at ablating cardiac sympathetic neurons and reducing resting, exercise, and reflex sympathetic activity. Additional studies are required to further characterize the physiological responses to this procedure as well as determine if this new approach is safe and efficacious for the treatment of conditions associated with excess sympathetic activity (e.g., autonomic dysreflexia, hypertension, heart failure, and ventricular arrhythmias). PMID- 19304950 TI - Aspergillus alabamensis, a new clinically relevant species in the section Terrei. AB - Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins enolase (enoA), beta-tubulin (benA), and calmodulin (calM) of a large number of isolates within the section Terrei, genus Aspergillus, revealed the presence of a new cryptic species within this section, Aspergillus alabamensis. Most members of this new cryptic species were recovered as colonizing isolates from immunocompetent patient populations, had decreased in vitro susceptibilities to the antifungal drug amphotericin B, and were morphologically similar to but genetically distinct from Aspergillus terreus isolates. PMID- 19304951 TI - RNA interference-mediated silencing of ornithine decarboxylase and spermidine synthase genes in Trypanosoma brucei provides insight into regulation of polyamine biosynthesis. AB - Polyamine biosynthesis is a drug target for the treatment of African sleeping sickness; however, mechanisms regulating the pathway in Trypanosoma brucei are not well understood. Recently, we showed that RNA interference (RNAi)-mediated gene silencing or the inhibition of S-adenosylmethionine decarboxylase (AdoMetDC) led to the upregulation of the AdoMetDC activator, prozyme, and ornithine decarboxylase (ODC) proteins. To determine if this regulatory response is specific to AdoMetDC, we studied the effects of the RNAi-induced silencing of the spermidine synthase (SpdSyn) and ODC genes in bloodstream form T. brucei. The knockdown of either gene product led to the depletion of the polyamine and trypanothione pools and to cell death. Decarboxylated AdoMet levels were elevated, while AdoMet was not affected. There was no significant effect on the protein levels of other polyamine pathway enzymes. The treatment of parasites with the ODC inhibitor alpha-difluoromethylornithine gave similar results to those observed for ODC knockdown. Thus, the cellular response to the loss of AdoMetDC activity is distinctive, suggesting that AdoMetDC activity controls the expression levels of the other spermidine biosynthetic enzymes. RNAi-mediated cell death occurred more rapidly for ODC than for SpdSyn. Further, the ODC RNAi cells were rescued by putrescine, but not spermidine, suggesting that the depletion of both putrescine and spermidine is more detrimental than the depletion of spermidine alone. This finding may contribute to the effectiveness of ODC as a target for the treatment of African sleeping sickness, thus providing important insight into the mechanism of action of a key antitrypanosomal agent. PMID- 19304952 TI - Oxidative stress function of the Saccharomyces cerevisiae Skn7 receiver domain. AB - The bifunctional Saccharomyces cerevisiae Skn7 transcription factor regulates osmotic stress response genes as well as oxidative stress response genes; however, the mechanisms involved in these two types of regulation differ. Skn7 osmotic stress activity depends on the phosphorylation of the receiver domain aspartate, D427, by the Sln1 histidine kinase. In contrast, D427 and the SLN1 SKN7 phosphorelay are dispensable for the oxidative stress response, although the receiver domain is required. The majority of oxidative stress response genes regulated by Skn7 also are regulated by the redox-responsive transcription factor Yap1. It is therefore possible that the nuclearly localized Skn7 does not itself respond to the oxidant but simply cooperates with Yap1 when it translocates to the nucleus. We report here that oxidative stress leads to a phosphatase sensitive, slow-mobility Skn7 variant. This suggests that Skn7 undergoes a posttranslational modification by phosphorylation following exposure to oxidant. Oxidant-dependent Skn7 phosphorylation was eliminated in strains lacking the Yap1 transcription factor. This suggests that the phosphorylation of Skn7 is regulated by Yap1. Mutations in the receiver domain of Skn7 were identified that affect its oxidative stress function. These mutations were found to compromise the association of Yap1 and Skn7 at oxidative stress response gene promoters. A working model is proposed in which the association of Yap1 with Skn7 in the nucleus is a prerequisite for Skn7 phosphorylation and the activation of oxidative stress response genes. PMID- 19304953 TI - Overexpression of astrocyte elevated gene-1 (AEG-1) is associated with esophageal squamous cell carcinoma (ESCC) progression and pathogenesis. AB - Astrocyte elevated gene-1 (AEG-1), upregulated in various types of human cancers, has been reported to be associated with the carcinogenesis of human cancer. However, the functional significance of AEG-1 in human esophageal squamous cell carcinoma (ESCC) remains unknown. In the present study, we showed the expression of AEG-1 was markedly upregulated in esophageal cancer cell lines and surgical ESCC specimens at both transcriptional and translational levels. Immunohistochemical analysis revealed that 80 of 168 (47.6%) paraffin-embedded archival ESCC specimens exhibited high levels of AEG-1 expression. Statistical analysis suggested the upregulation of AEG-1 was significantly correlated with the clinical staging of the ESCC patients (P = 0.001), T classification (P = 0.002), N classification (P = 0.034), M classification (P = 0.021) and histological differentiation (P = 0.035) and those patients with high AEG-1 levels exhibited shorter survival time (P < 0.001). Multivariate analysis indicated that AEG-1 expression might be an independent prognostic indicator of the survival of patients with ESCC. Furthermore, we found that ectopic expression of AEG-1 in ESCC cells could significantly enhance cell proliferation and anchorage-independent growth ability. Conversely, silencing AEG-1 by short hairpin RNAi caused an inhibition of cell growth and anchorage-independent growth ability on soft agar. Moreover, we demonstrated that the upregulation of AEG-1 could reduce the expression of p27(Kip1) and induce the expression of cyclin D1 through the AKT/FOXO3a pathway. Our findings suggest that the AEG-1 protein is a valuable marker of ESCC progression and that the upregulation of AEG-1 plays an important role in the development and pathogenesis of human ESCC. PMID- 19304954 TI - Zebrafish mutants in the von Hippel-Lindau tumor suppressor display a hypoxic response and recapitulate key aspects of Chuvash polycythemia. AB - We have generated 2 zebrafish lines carrying inactivating germline mutations in the von Hippel-Lindau (VHL) tumor suppressor gene ortholog vhl. Mutant embryos display a general systemic hypoxic response, including the up-regulation of hypoxia-induced genes by 1 day after fertilization and a severe hyperventilation and cardiophysiologic response. The vhl mutants develop polycythemia with concomitantly increased epo/epor mRNA levels and erythropoietin signaling. In situ hybridizations reveal global up-regulation of both red and white hematopoietic lineages. Hematopoietic tissues are highly proliferative, with enlarged populations of c-myb(+) hematopoietic stem cells and circulating erythroid precursors. Chemical activation of hypoxia-inducible factor signaling recapitulated aspects of the vhl(-/-) phenotype. Furthermore, microarray expression analysis confirms the hypoxic response and hematopoietic phenotype observed in vhl(-/-) embryos. We conclude that VHL participates in regulating hematopoiesis and erythroid differentiation. Injections with human VHLp30 and R200W mutant mRNA demonstrate functional conservation of VHL between mammals and zebrafish at the amino acid level, indicating that vhl mutants are a powerful new tool to study genotype-phenotype correlations in human disease. Zebrafish vhl mutants are the first congenital embryonic viable systemic vertebrate animal model for VHL, representing the most accurate model for VHL-associated polycythemia to date. They will contribute to our understanding of hypoxic signaling, hematopoiesis, and VHL-associated disease progression. PMID- 19304955 TI - Comparative ability of IL-12 and IL-28B to regulate Treg populations and enhance adaptive cellular immunity. AB - Improving the potency of immune responses is paramount among issues concerning vaccines against deadly pathogens. IL-28B belongs to the newly described interferon lambda (IFNlambda) family of cytokines, and has not yet been assessed for its potential ability to influence adaptive immune responses or act as a vaccine adjuvant. We compared the ability of plasmid-encoded IL-28B to boost immune responses to a multiclade consensus HIV Gag plasmid during DNA vaccination with that of IL-12. We show here that IL-28B, like IL-12, is capable of robustly enhancing adaptive immunity. Moreover, we describe for the first time how IL-28B reduces regulatory T-cell populations during DNA vaccination, whereas IL-12 increases this cellular subset. We also show that IL-28B, unlike IL-12, is able to increase the percentage of splenic CD8(+) T cells in vaccinated animals, and that these cells are more granular and have higher antigen-specific cytolytic degranulation compared with cells taken from animals that received IL-12 as an adjuvant. Lastly, we report that IL-28B can induce 100% protection from mortality after a lethal influenza challenge. These data suggest that IL-28B is a strong candidate for further studies of vaccine or immunotherapy protocols. PMID- 19304956 TI - KIR acquisition probabilities are independent of self-HLA class I ligands and increase with cellular KIR expression. AB - Inhibitory killer cell immunoglobulin-like receptors (KIRs) preserve tolerance to self and shape the functional response of human natural killer (NK) cells. Here, we have evaluated the influence of selection processes in the formation of inhibitory KIR repertoires in a cohort of 44 donors homozygous for the group A KIR haplotype. Coexpression of multiple KIRs was more frequent than expected by the product rule that describes random association of independent events. In line with this observation, the probability of KIR acquisition increased with the cellular expression of KIRs. Three types of KIR repertoires were distinguished that differed in frequencies of KIR- and NKG2A-positive cells but showed no dependency on the number of self-HLA class I ligands. Furthermore, the distribution of self- and nonself-KIRs at the cell surface reflected a random combination of receptors rather than a selection process conferred by cognate HLA class I molecules. Finally, NKG2A was found to buffer overall functional responses in KIR repertoires characterized by low-KIR expression frequencies. The results provide new insights into the formation of inhibitory KIR repertoires on human NK cells and support a model in which variegated KIR repertoires are generated through sequential and random acquisition of KIRs in the absence of selection. PMID- 19304957 TI - Prevalence and prognostic implications of CEBPA mutations in pediatric acute myeloid leukemia (AML): a report from the Children's Oncology Group. AB - CEBPA mutations have been associated with improved outcome in adult acute myeloid leukemia (AML). We evaluated the prevalence and prognostic significance of CEBPA mutations in 847 children with AML treated on 3 consecutive pediatric trials. Two types of CEBPA mutations-N-terminal truncating mutations and in-frame bZip-domain mutations-were detected in 38 (4.5%) of 847 patients tested; 31 (82%) of 38 patients with mutations harbored both mutation types. Mutation status was correlated with laboratory and clinical characteristics and clinical outcome. CEBPA mutations were significantly more common in older patients, patients with FAB M1 or M2, and patients with normal karyotype. Mutations did not occur in patients with either favorable or unfavorable cytogenetics. Actuarial event-free survival at 5 years was 70% versus 38% (P = .015) with a cumulative incidence of relapse from complete remission of 13% versus 44% (P = .007) for those with and without CEBPA mutations. The presence of CEBPA mutations was an independent prognostic factor for improved outcome (HR = 0.24, P = .047). As CEBPA mutations are associated with lower relapse rate and improved survival, CEBPA mutation analysis needs to be incorporated into initial screening for risk identification and therapy allocation at diagnosis. PMID- 19304958 TI - Characterization of novel and complex genomic aberrations in glioblastoma using a 32K BAC array. AB - Glioblastomas (GBs) are malignant CNS tumors often associated with devastating symptoms. Patients with GB have a very poor prognosis, and despite treatment, most of them die within 12 months from diagnosis. Several pathways, such as the RAS, tumor protein 53 (TP53), and phosphoinositide kinase 3 (PIK3) pathways, as well as the cell cycle control pathway, have been identified to be disrupted in this tumor. However, emerging data suggest that these aberrations represent only a fraction of the genetic changes involved in gliomagenesis. In this study, we have applied a 32K clone-based genomic array, covering 99% of the current assembly of the human genome, to the detailed genetic profiling of a set of 78 GBs. Complex patterns of aberrations, including high and narrow copy number amplicons, as well as a number of homozygously deleted loci, were identified. Amplicons that varied both in number (three on average) and in size (1.4 Mb on average) were frequently detected (81% of the samples). The loci encompassed not only previously reported oncogenes (EGFR, PDGFRA, MDM2, and CDK4) but also numerous novel oncogenes as GRB10, MKLN1, PPARGC1A, HGF, NAV3, CNTN1, SYT1, and ADAMTSL3. BNC2, PTPLAD2, and PTPRE, on the other hand, represent novel candidate tumor suppressor genes encompassed within homozygously deleted loci. Many of these genes are already linked to several forms of cancer; others represent new candidate genes that may serve as prognostic markers or even as therapeutic targets in the future. The large individual variation observed between the samples demonstrates the underlying complexity of the disease and strengthens the demand for an individualized therapy based on the genetic profile of the patient. PMID- 19304959 TI - PTPmu suppresses glioma cell migration and dispersal. AB - The cell-surface receptor protein tyrosine phosphatase mu (PTPmu) is a homophilic cell adhesion molecule expressed in CNS neurons and glia. Glioblastomas (GBMs) are the highest grade of primary brain tumors with astrocytic similarity and are characterized by marked dispersal of tumor cells. PTPmu expression was examined in human GBM, low-grade astrocytoma, and normal brain tissue. These studies revealed a striking loss of PTPmu protein expression in highly dispersive GBMs compared to less dispersive low-grade astrocytomas and normal brain. We hypothesized that PTPmu contributes to contact inhibition of glial cell migration by transducing signals in response to cell adhesion. Therefore, loss of PTPmu may contribute to the extensive dispersal of GBMs. The migration of brain tumor cells was assessed in vitro using a scratch wound assay. Parental U-87 MG cells express PTPmu and exhibited limited migration. However, short-hairpin RNA (shRNA) mediated knockdown of PTPmu induced a morphological change and increased migration. Next, a brain slice assay replicating the three-dimensional environment of the brain was used. To assess migration, labeled U-87 MG glioma cells were injected into adult rat brain slices, and their movement was followed over time. Parental U-87 MG cells demonstrated limited dispersal in this assay. However, PTPmu shRNA induced migration and dispersal of U-87 MG cells in the brain slice. Finally, in a mouse xenograft model of intracranially injected U-87 MG cells, PTPmu shRNA induced morphological heterogeneity in these xenografts. Together, these data suggest that loss of PTPmu in human GBMs contributes to tumor cell migration and dispersal, implicating loss of PTPmu in glioma progression. PMID- 19304960 TI - Pervasive, genome-wide positive selection leading to functional divergence in the bacterial genus Campylobacter. AB - An open question in bacterial genomics is the role that adaptive evolution of the core genome plays in diversification and adaptation of bacterial species, and how this might differ between groups of bacteria occupying different environmental circumstances. The genus Campylobacter encompasses several important human and animal enteric pathogens, with genome sequence data available for eight species. We estimate the Campylobacter core genome at 647 genes, with 92.5% of the nonrecombinant core genome loci under positive selection in at least one lineage and the same gene frequently under positive selection in multiple lineages. Tests are provided that reject recombination, saturation, and variation in codon usage bias as factors contributing to this high level of selection. We suggest this genome-wide adaptive evolution may result from a Red Queen macroevolutionary dynamic, in which species are involved in competition for resources within the mammalian and/or vertebrate gastrointestinal tract. Much reduced levels of positive selection evident in Streptococcus, as reported by the authors in an earlier work, may be a consequence of these taxa inhabiting less species-rich habitats, and more unique niches. Despite many common loci under positive selection in multiple Campylobacter lineages, we found no evidence for molecular adaptive convergence at the level of the same or adjacent codons, or even protein domains. Taken collectively, these results describe the diversification of a bacterial genus that involves pervasive natural selection pressure across virtually the entire genome, with this adaptation occurring in different ways in different lineages, despite the species tendency toward a common gastrointestinal habitat. PMID- 19304961 TI - Angiocentric glioma-induced seizures in a 2-year-old child. AB - A 2-year-old child presented with medically refractory seizures and was found to have a right frontoparietal parasagittal angiocentric glioma. Depth electrodes were used to document ictal onset from within the tumor rather than from the surrounding tissues. Ictal activity then spread to a wide area on the cortical surface, including the region around the tumor and hand motor cortex. Lesionectomy permitted sparing of adjacent areas of eloquent cortex, and the child is now seizure-free on monotherapy. PMID- 19304962 TI - Immune reconstitution inflammatory syndrome. AB - Antiretroviral therapy (ART) inhibits HIV replication, allowing recovery of CD4+ T cell numbers and the restoration of immune function; its introduction has led to improved outcomes for individuals with HIV infection. However, it has been observed that some individuals responding to ART experience a clinical deterioration with symptoms and signs of an inflammatory illness. Immune reconstitution inflammatory syndrome (IRIS) results from pathological immune responses occurring during immune reconstitution. IRIS is best considered a group of disorders with a wide range of clinical manifestations, incorporating disease resulting from pathological inflammation to pathogens, immune-mediated inflammatory disease and autoimmune disease. Clinical effects range from a mild, self-limiting illness to severe morbidity and mortality. Clinicians working in the field of HIV medicine can expect to encounter individuals with IRIS. In this review, we discuss definitions, describe clinical presentations, summarize research relating to pathogenesis and identify risk factors, preventive and management strategies. PMID- 19304963 TI - Suboptimal therapy and clinical management of gonorrhoea in an area with high level antimicrobial resistance. AB - The choice of antimicrobial agents for the treatment of gonorrhoea is critical in areas where the prevalence of drug resistance is high. This study aimed to evaluate the antibiotic treatment of gonorrhoea in endemic areas. During 1999 2004, all Neisseria gonorrhoeae infections (n = 90) were evaluated. Patients' medical records and antibiotic treatment regimens were retrospectively reviewed if their isolates were viable (n = 65). In vitro antimicrobial susceptibility of N. gonorrhoeae isolates was performed. Urethritis (89%) and pelvic inflammatory disease (42%) were the most common presentations among men (n = 53) and women (n = 12), respectively. Of 54 patients with uncomplicated N. gonorrhoeae infection, 32 of them received appropriate antibiotics, including cefuroxime (n = 20), ceftriaxone (n = 10), ciprofloxacin (n = 1) and azithromycin (n = 1) during follow-ups. Among 65 patients, 53.8% were notified to the health authority. Check ups of other sexually transmitted diseases were carried out in only 46% of patients. Not all isolates were susceptible to penicillin, 96.9% were resistant to tetracycline and 86% were resistant to ciprofloxacin. Ceftriaxone, cefixime, spectinomycin and azithromycin were active in vitro against all isolates. In conclusion, It is crucial to develop treatment guidelines according to regional antimicrobial resistances and educational programmes to improve clinical care for genital gonococcal diseases. PMID- 19304964 TI - Should syphilis be treated differently in HIV-positive and HIV-negative individuals? Treatment outcomes at a university hospital, Brighton, UK. AB - There has been much debate regarding the optimum treatment for syphilis in HIV positive patients. There has been a shift in expert opinion in the UK towards using two doses of benzathine penicillin G one week apart regardless of HIV status. We report our experience using a 17-day course of daily procaine penicillin plus probenecid in HIV-positive individuals and two doses of benzathine in HIV-negative patients. Of 350 cases of early syphilis, 37% were in HIV-positive individuals. Ninety-eight percent of HIV-positive patients completing the treatment were followed up for at least six months and met the criteria for treatment success. The treatment response was equally good (98%) for HIV-negative patients using these different schedules. It is currently unclear which patients require an extended course of treatment for syphilis. We have demonstrated that patients adhere well to this regimen, and significantly we have shown comparable treatment success rates in HIV-positive and -negative individuals. PMID- 19304965 TI - The Internet as a diagnostic aid: the patients' perspective. AB - A questionnaire-based cross-sectional study was conducted in order to determine how frequently patients attending a genitourinary (GU) medicine clinic use the Internet to diagnose their own symptoms, and to assess the accuracy of their diagnosis. Out of 223 symptomatic patients attending a GU clinic, 101 (45.3%) looked up their symptoms on the Internet. The age difference between those who looked up their symptoms and those who did not was not statistically significant (P = 0.77). Twenty (19.8%) out of 101 patients diagnosed their own symptoms, and 14 (13.9%) patients made the correct diagnosis. Ninety-one (90.1%) patients used the Google((R)) search engine as the starting point for their search. Although the Internet plays an important role in providing health information, the variable quality of health information available limits its use as a diagnostic tool by patients. PMID- 19304966 TI - Association of Mycoplasma genitalium with acute non-gonococcal urethritis in Russian men: a comparison with gonococcal and chlamydial urethritis. AB - Urethral specimens from 172 men who attended sexually transmitted disease clinics in the Moscow Oblast were examined for Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium by nucleic acid amplification tests. N. gonorrhoeae was detected in the urethra of 41 (24%) of the 172 men and C. trachomatis in 57 (33%). The latter occurred in 15 (36%) of the 41 men who were infected by N. gonorrhoeae and in 42 (32%) of 131 uninfected by gonococci. Of the 42 men uninfected by gonococci but chlamydia infected, 39 (93%) had symptoms and/or signs of urethritis. M. genitalium was detected in 45 (26%) of the 172 men, in nine (22%) of the 41 men infected with N. gonorrhoeae and in 12 (21%) infected with C. trachomatis. M. genitalium was detected alone in 25 (28%) of the 89 men uninfected by either gonococci or C. trachomatis. Of these 25 men, 24 (96%) had urethral symptoms and signs of inflammation, a proportion significantly more than experienced by the 64 men uninfected by any of the microorganisms. Of the 31 men who apparently had no symptoms or signs of urethritis, only three (10%) were infected by M. genitalium. The data provide evidence for the pathogenicity and frequent occurrence of M. genitalium in men in Moscow and presumably elsewhere in Russia. PMID- 19304967 TI - Haemophilus ducreyi detection by polymerase chain reaction in oesophageal lesions of HIV patients. AB - HIV patients frequently have opportunistic oesophageal infections. We report Haemophilus ducreyi genetic material detected by polymerase chain reaction in biopsies of oesophageal lesions in three HIV-1-infected patients. This finding may be an indication of its aetiopathological role in oesophageal lesions of HIV patients. PMID- 19304968 TI - Predictors for casual sex and/or infection among sexually transmitted disease clinic attendees in China. AB - To assess the risk factors for casual sex and infections among the sexually transmitted disease (STD) clinic attendees in two disparate Chinese cities, an STD clinic-based cross-sectional study was conducted to provide demographic and sexual behaviour information. Participants were recruited from nine STD clinics selected by mapping strategy. STD prevalence was 69.4% (68.6% of men and 65.2% of women). The most common diagnoses were non-gonococcal urethritis (22.2%), genital warts (13.2%), syphilis (11.6%), gonorrhoea (8.4%), chlamydia (6.3%) and herpes simplex virus type 2 (HSV-2) (5.8%). Of 536 participants, 22.5% reported having casual sex in the last three months, younger age, less education, unawareness of transmission routes and having had casual sex in the last three months were independent risk factors for acquisition of an STD. Single or separated marital status, non-local residency and STD diagnoses were independently associated with having had casual sex. After decades of exceedingly low STD rates in China, a full panoply of STD diagnoses are now evident. Both for reproductive health concerns and for stemming the expansion of HIV spread, STD control and prevention must be revitalized as a priority for China's public health and medical institutions. Effective training is a priority, given the dearth of STD experienced health-care workers. PMID- 19304969 TI - Resistance mutations in HIV-1 infected pregnant women and their infants receiving antiretrovirals to prevent HIV-1 vertical transmission in China. AB - The objective of this study was to assess the resistance baseline in HIV-1 infected pregnant women in China and the emergence of drug resistance (DR) among them and their infants after receiving single-dose nevirapine (sdNVP) or zidovudine plus sdNVP (ZDV-sdNVP) for the prevention of mother-to-child transmission (PMTCT). The prospective study followed 62 drug-naive HIV-1-infected mothers and their infants who received sdNVP for PMTCT and 18 who received ZDV sdNVP. Primary DR was detected in 8.8% (7/80) of pregnant women at baseline. Two (2.5%) women had mutations associated with nucleoside reverse transcriptase inhibitors (NRTI). Six (7.5%) women harboured mutations associated with non nucleoside reverse transcriptase inhibitors (NNRTI). No protease inhibitor (PI) DR was detected. The DR rates in infected mothers selected by sdNVP and ZDV-sdNVP regimen were 16.3% and 0%, respectively. Women with the emergence of DR after sdNVP had lower CD4 cell counts at delivery than women without DR after sdNVP (178 vs.364 cells/mm(3), P < 0.05). The vertical transmission rate in women with baseline resistance was similar to that in women without baseline resistance (1/7 vs. 3/73, P > 0.05). The study indicates that NVP resistance after sdNVP was associated with CD4 cell count at delivery. ZDV-sdNVP regimen was of more significance in the prevention of the emergence of NNRTI-related DR than sdNVP. PMID- 19304970 TI - Loneliness, social support and family function of people living with HIV/AIDS in Anhui rural area, China. AB - The main purpose of this study was to investigate how loneliness was associated with social support and family function among people living with HIV/AIDS (PLWHA) in an HIV-stricken area of China. Two hundred and nineteen PLWHA were surveyed using the UCLA (University of California at Los Angeles)-Loneliness Scale, the Social Support Rate Scale and the Family APGAR (Adaptability, Partnership, Growth, Affection and Resolve) Index. The results indicated that the majority (84.5%) of PLWHA had moderate to high levels of loneliness, with a mean score of 47.62 and a standardized score of 59.53. The level of loneliness was significantly different in subjects' occupations, but not in other sociodemographic variables. Social support, family function and all dimensions except utilizations of support were significantly negatively associated with loneliness. Multiple regression revealed that less social support and poor family function were associated with more loneliness. In conclusion, loneliness prevails among PLWHA. It may limit PLWHA's ability or access to social relationship. These findings support the hypothesis that if PLWHA are better supported and cared for, their negative psychosocial consequences might be prevented or at least reduced. PMID- 19304971 TI - Identification of oral candidosis, hairy leukoplakia and recurrent oral ulcers as distinct cases of immune reconstitution inflammatory syndrome. AB - Oral lesions such as candidosis, hairy leukoplakia (HL) and oral ulcers are strikingly absent in the numerous reports of immune reconstitution inflammatory syndrome (IRIS). To document oral manifestations attributable to immune reconstitution, we conducted a longitudinal follow-up of a cohort of HIV+ individuals starting highly active antiretroviral therapy (HAART) and completing oral pathology follow-up up to 12 weeks after treatment initiation. HIV-infected patients had oral examinations, CD4+ T-cell count and viral load determinations performed at baseline, and at weeks 4, 8 and 12 after HAART initiation. Among individuals with satisfactory viral response and recovery of > or =50 CD4+ T cell/microL, eight patients complied with strict IRIS criteria: two developed clinical signs of oral candidosis (OC), two oral ulcers, three HL and one Kaposi's sarcoma. CD4+ T-cell counts at symptom onset suggested no remaining immune suppression. Our findings show that cases of OC, HL and recurrent ulcers can be instances of IRIS. PMID- 19304972 TI - Effect of HIV viral load, CD4 cell count and antiretroviral therapy on human papillomavirus prevalence in urine samples of HIV-infected men. AB - HIV-infected patients are at increased risk for persistent human papillomavirus (HPV) infection, the major cause of anogenital cancer. The present study describes the HPV prevalence in urine samples of 243 HIV-infected men and a control group of 231 men. HPV DNA was amplified by the SPF10 polymerase chain reaction primer set. The overall HPV prevalence in HIV-infected men was 27.5% compared with 12.6% in controls (P < 0.01). Infections with high-risk and multiple HPV genotypes were present in both groups. Differences were not statistically significant. A multivariate logistic regression model showed a decreased HPV prevalence associated with use of a nucleoside and a non-nucleoside reverse transcriptase inhibitor combination (P = 0.03). A trend was observed towards a higher HPV prevalence and a lower CD4 cell count. Further prospective studies are needed to determine the role of HPV DNA testing in urine in future screening programmes for anal cancer in men. PMID- 19304973 TI - Time taken to undetectable viral load, following the initiation of HAART. AB - The notes of all HIV patients in Sheffield, registered at the Department of Genitourinary Medicine at the Royal Hallamshire Hospital, who had initiated HAART in 2005 and 2006, were audited. The aim was to determine what percentage of these patients acquired an undetectable viral load within 24 weeks and whether this was greater than 75% in accordance with the BHIVA guidelines. Twenty-nine (78.4%) of the 37 patients who were initiating treatment for the first time had an undetectable viral load after 24 weeks. PMID- 19304974 TI - How many cases of chlamydial infection would we miss by not testing partners for infection? AB - Quality Improvement Scotland standards for sexual health services require that 0.64 contacts per case should be verified as having attended within 90 days of the first partner notification interview. Partner delivered therapy results in more patients being treated than when patients are given information for partners but removes the possibility of further cases of Chlamydia trachomatis being diagnosed through the tracing of secondary contacts. An audit was performed to estimate the impact of removing secondary contacts on the number of chlamydial infections identified in our service. Patients who were not known to be contacts of chlamydial infection were included. One hundred and twenty-seven index cases generated 189 contacts, of which 100 were confirmed as tested and treated. Sixty four were C. trachomatis positive, who in turn generated 36 new contacts. Fourteen of these were positive. Secondary contact tracing identified 22-28% more cases of chlamydial infection in our service than if all partners were treated without testing. PMID- 19304975 TI - Documentation of advice regarding safer sexual practices and partner notification within an HIV clinic. AB - Guidelines for the sexual health care of our HIV patients and provision of post exposure prophylaxis following sexual exposure have been produced by both British Association of Sexual Health & HIV and British HIV Association (BHIVA). In the light of recent criminal prosecutions, BHIVA produced guidance for HIV clinical teams regarding discussions with patients about HIV transmission and reducing the risk to sexual partners. This audit examined the advice given to HIV patients with regard to safer sexual practices, sexually transmitted infection screening and partner notification and found that, on the whole, the standards set by national guidelines were adhered to, although areas for improvement were identified. We hope that the introduction of proformas with specific prompts for these subjects for HIV clinic visits will improve clinical standards in this area. PMID- 19304976 TI - Unusual late nodular presentation of secondary syphilis. AB - A 20-year-old man presented with a five-week history of an eruption of papules and nodules disseminated over his body and face. We propose that this patient has a late form of secondary syphilis with a nodular, granulomatous inflammation in urgent need of treatment. Otherwise late irreversible sequelae could develop and unwanted possible further sexual transmission could take place. PMID- 19304977 TI - Recurrence of cryptococcal meningitis in HIV-infected patients following immune reconstitution. AB - Two HIV-infected patients had recurrent cryptococcal meningitis (CM) despite treatment with fluconazole and immune reconstitution with combination antiretroviral therapy (CART). Following treatment of CM with fluconazole, lumbar puncture should be performed either after completion of induction treatment for CM or before starting CART, in order to confirm cerebrospinal fluid sterility. PMID- 19304978 TI - Abacavir-induced reaction with fever and severe skin rash in a patient tested human leukocyte antigen-B*5701 negative. AB - The most serious adverse event caused by abacavir is the hypersensitivity reaction, which is usually associated with the presence of the human leukocyte antigen (HLA) subtype B*5701, as shown in recent studies. We describe the case of a 41-year-old Caucasian female patient, who tested HLA-B*5701 negative and developed fever and severe skin rash 10 weeks after the start of abacavir therapy. Similar reports suggest that not all severe abacavir-induced adverse events occur as a result of classic hypersensitivity reactions, and can present also in HLA-B*5701-negative patients. PMID- 19304980 TI - Chylous ascites: a late complication of intra-abdominal Mycobacterium avium complex immune reconstitution syndrome in HIV-infected patients. AB - Chylous ascites related to Mycobacterium avium complex (MAC) in HIV-infected patients is rare, with only six cases reported in the English literature. We report a series of six cases from a single institution. During the past six years, chylous ascites was diagnosed in six (35%) of 17 AIDS patients, all of whom had previously been diagnosed with intra-abdominal MAC immune reconstitution syndrome (MAC-IRS). A review of medical records identified no other cases of chylous ascites among HIV-positive patients over the past 13 years (1994-2007), and the incidence was estimated at one in 2248 HIV-positive admissions. The ascitic fluid had a milky appearance and a median triglyceride level of 4.07 mmol/L (range 3.19-29.6 mmol/L) (360 mg/dL, range 282-2620 mg/dL). After a median follow-up of 20 months, five (83%) of six patients survived. Chylous ascites is a late complication of intra-abdominal MAC-IRS, and is usually associated with a favourable prognosis. PMID- 19304979 TI - Syphilitic hepatitis among HIV-infected patients. AB - Syphilis is an important public health issue and continues to occur at high rates among HIV-infected patients. Although abnormal liver function tests are common among HIV-infected persons, the incidence of syphilitic hepatitis in this population is currently unknown. We present two cases of syphilitic hepatitis and performed a retrospective study to determine the incidence of hepatitis during early syphilis infections among HIV-infected persons. Our study showed that syphilitic hepatitis is common, occurring in 38% (12/32) of HIV-positive patients with early stages of syphilis infection. Most cases occurred during secondary syphilis, with the most common finding being a maculopapular rash. Syphilis should be included in the differential diagnosis of HIV patients presenting with liver test abnormalities, rash and/or sexual risk factors. PMID- 19304981 TI - A delayed hypersensitivity reaction to enfuvirtide after rechallenge. AB - Enfuvirtide is beneficial in patients with limited treatment options. We report this case to highlight the possibility of a delayed hypersensitivity reaction as an important potential side-effect of enfuvirtide treatment. A highly antiretroviral treatment-experienced man was commenced on a new regimen containing enfuvirtide. Prophylaxis for Pneumocystis jirovecii pneumonia was started using trimethoprim/sulphamethoxazole (TMP-STX) simultaneously. Ten days later, he developed a maculopapular rash on the chest and abdomen without any systemic features. Both enfuvirtide and TMP-STX were discontinued. Re introduction of enfuvirtide occurred in a hospital setting. Before re-challenge, haemodynamic observations were stable. The rash re-appeared involving the whole body 5 hours post-dose and was associated with fever (temperature 38.4), nausea and a presyncopal episode. Hypersensitivity to this drug occurred immediately post-dose in phase III trials. Enfuvirtide is a useful drug in those with reduced drug options. The possibility of delayed hypersensitivity has not been reported previously. PMID- 19304982 TI - The HPV vaccine for men. PMID- 19304983 TI - Microscopy in asymptomatic men. PMID- 19304984 TI - Atypical presentation of secondary syphilis in a psoriatic HIV patient. PMID- 19304985 TI - Quality enhancements of laboratory diagnosis of sexually transmitted infections in Russia. PMID- 19304986 TI - The neuromediator glutamate, through specific substrate interactions, enhances mitochondrial ATP production and reactive oxygen species generation in nonsynaptic brain mitochondria. AB - The finding that upon neuronal activation glutamate is transported postsynaptically from synaptic clefts and increased lactate availability for neurons suggest that brain mitochondria (BM) utilize a mixture of substrates, namely pyruvate, glutamate, and the tricarboxylic acid cycle metabolites. We studied how glutamate affected oxidative phosphorylation and reactive oxygen species (ROS) production in rat BM oxidizing pyruvate + malate or succinate. Simultaneous oxidation of glutamate + pyruvate + malate increased state 3 and uncoupled respiration by 52 and 71%, respectively. The state 4 ROS generation increased 100% over BM oxidizing pyruvate + malate and 900% over that of BM oxidizing glutamate + malate. Up to 70% of ROS generation was associated with reverse electron transport. These effects of pyruvate + glutamate + malate were observed only with BM and not with liver or heart mitochondria. The effects of glutamate + pyruvate on succinate-supported respiration and ROS generation were not organ-specific and depended only on whether mitochondria were isolated with or without bovine serum albumin. With the non-bovine serum albumin brain and heart mitochondria oxidizing succinate, the addition of pyruvate and glutamate abrogated inhibition of Complex II by oxaloacetate. We conclude that (i) during neuronal activation, simultaneous oxidation of glutamate + pyruvate temporarily enhances neuronal mitochondrial ATP production, and (ii) intrinsic inhibition of Complex II by oxaloacetate is an inherent mechanism that protects against ROS generation during reverse electron transport. PMID- 19304987 TI - Hepatic triacylglycerol hydrolysis regulates peroxisome proliferator-activated receptor alpha activity. AB - Recent evidence suggests that fatty acids generated from intracellular triacylglycerol (TAG) hydrolysis may have important roles in intracellular signaling. This study was conducted to determine if fatty acids liberated from TAG hydrolysis regulate peroxisome proliferator-activated receptor alpha (PPARalpha). Primary rat hepatocyte cultures were treated with adenoviruses overexpressing adipose differentiation-related protein (ADRP) or adipose triacylglycerol lipase (ATGL) or treated with short interfering RNA (siRNA) targeted against ADRP. Subsequent effects on TAG metabolism and PPARalpha activity and target gene expression were determined. Overexpressing ADRP attenuated TAG hydrolysis, whereas siRNA-mediated knockdown of ADRP or ATGL overexpression resulted in enhanced TAG hydrolysis. Results from PPARalpha reporter activity assays demonstrated that decreasing TAG hydrolysis by ADRP overexpression resulted in a 35-60% reduction in reporter activity under basal conditions or in the presence of fatty acids. As expected, PPARalpha target genes were also decreased in response to ADRP overexpression. However, the PPARalpha ligand, WY-14643, was able to restore PPARalpha activity following ADRP overexpression. Despite its effects on PPARalpha, overexpressing ADRP did not affect PPARgamma activity. Enhancing TAG hydrolysis through ADRP knockdown or ATGL overexpression increased PPARalpha activity. These results indicate that TAG hydrolysis and the consequential release of fatty acids regulate PPARalpha activity. PMID- 19304988 TI - Homelessness as an independent risk factor for mortality: results from a retrospective cohort study. AB - BACKGROUND: Homelessness is associated with increased risks of mortality but it has not previously been possible to distinguish whether this is typical of other socio-economically deprived populations, the result of a higher prevalence of morbidity or an independent risk of homelessness itself. The aim of this study was to describe mortality among a cohort of homeless adults and adjust for the effects of morbidity and socio-economic deprivation. METHODS: Retrospective 5 year study of two fixed cohorts, homeless adults and an age- and sex-matched random sample of the local non-homeless population in Greater Glasgow National Health Service Board area for comparison. RESULTS: Over 5 years of observation, 1.7% (209/12 451) of the general population and 7.2% (457/6323) of the homeless cohort died. The hazard ratio of all-cause mortality in homeless compared with non-homeless cohorts was 4.4 (95% CI: 3.8-5.2). After adjustment for age, sex and previous hospitalization, homelessness was associated with an all-cause mortality hazard ratio of 1.6 (95% CI: 1.3-1.9). Homelessness had differential effects on cause-specific mortality. Among patients who had been hospitalized for drug related conditions, the homeless cohort experienced a 7-fold increase in risk of death from drugs compared with the general population. CONCLUSIONS: Homelessness is an independent risk factor for deaths from specific causes. Preventive programmes might be most effectively targeted at the homeless with these conditions. PMID- 19304989 TI - An innocent bystander in the coronary tree. PMID- 19304990 TI - Big men and atrial fibrillation: effects of body size and weight gain on risk of atrial fibrillation in men. AB - AIMS: Obesity is a recognized risk factor for atrial fibrillation (AF), partly because of the association between body mass index (BMI) and atrial volume. We aimed to determine whether other factors relating to body size were related to AF. METHODS AND RESULTS: Data were derived from a random population sample of 6903 men (mean age 51.5 years) who underwent a single midlife evaluation as part of the multifactor Swedish Primary Prevention Study. A total of 1253 men (18.2%) had a subsequent hospital discharge diagnosis (principal or secondary) of AF during a maximum follow-up of 34.3 years. Body surface area (BSA) at age 20 (calculated from recalled weight and measured height) was strongly related to subsequent AF (P < 0.0001), as were midlife BMI and weight gain from age 20 to midlife (P < 0.0001). In a Cox regression model which adjusted for midlife BMI, weight gain and other risk factors, hazard ratios (HR) [95% confidence intervals (CI)] for AF for the second, third, and fourth quartile of BSA at age 20, compared with the lowest quartile, were 1.47 (95% CI, 1.22-1.76), 1.66 (95% CI, 1.38-2.00), and 2.22 (95% CI, 1.82-2.70) (P for trend <0.0001). CONCLUSION: Large body size in youth, in an era when obesity was rare, as well as weight gain from age 20 to midlife, were both independently related to the development of AF. Given the current trends not only for obesity but also for height, a substantial increase in the incidence of AF is likely. PMID- 19304991 TI - Understanding suicidality and correlates among Chinese secondary school students in Hong Kong. AB - Suicide has become a leading cause of mortality and morbidity for adolescents in Hong Kong. This study investigated the factors associated with suicidal ideation and attempt among the secondary school students in Hong Kong by studying a representative sample of 3383 students with a self-administered questionnaire and analysed by multiple logistic regressions analysis. Youth risk behaviours, such as heavy smoking, episodic heavy alcohol drinking, early sexual experience, and feeling hopeless, were found to be associated with both suicidal thoughts and attempts, with 'misuse of drugs' as discriminating factor that solely related to attempt and 'involvement in physical fight' solely related to suicidal thoughts. Addition of suicidal ideation is a significant explanatory variable of suicidal attempt over and above health risk behaviours. Stratified analysis of upper and lower secondary students would give better understanding of significance of various risk factor for different age groups. The disturbing prevalence of suicidal behaviour and its coexistence with other high-risk behaviour in secondary school students have implications for teachers, youth workers and public health practitioners to develop and evaluate programmes for suicide prevention. PMID- 19304992 TI - Sexual behavior and drinking style among teenagers: a population-based study in Finland. AB - In this large-scale study, we examined the relationship between an adolescent's sexual behavior and drinking style. Three aspects of sexual risk-taking were included: early activity, unprotected sexual intercourse and having sex with multiple partners. A distinction was made between different drinking styles, i.e. alcohol drinking and drunkenness-related drinking. Cross-sectional school survey data from the School Health Promotion Study was collected in Finland in 2002 2003. The national sample consisted of adolescents from the eighth and ninth grades (n = 100,790). The mean ages were 14.8 and 15.8 years. Using logistic regression analysis, we investigated the association between sexual behavior and drinking style among teenagers. The likelihood of engaging in sexual intercourse increased with the frequency of alcohol use. In particular, frequent drunkenness related drinking increased the probability that the teenager had experienced sexual intercourse. The likelihood of engaging in unprotected sex and/or having multiple sexual partners was many-fold for adolescents drinking frequently until they were in a state of drunkenness. Particularly for girls, weekly drunkenness related drinking was associated with multiple partners. The vast majority of sexually experienced under-aged adolescents drink alcohol, many of them until they are drunk. Thus, it could be effective to combine both alcohol education and sex education, including contraceptive counseling, in early adolescence. PMID- 19304993 TI - Evaluation of male fertility potential by Toluidine Blue test for sperm chromatin structure assessment. AB - BACKGROUND: We have previously suggested that the Toluidine Blue (TB) test can be used for sperm chromatin structure assessment. In this study, we wished to evaluate the clinical applicability of the TB test in assessing male fertility potential using well-defined groups of fertile and infertile men. METHODS: Sixty three fertile and 79 infertile men were tested. Infertility thresholds for the proportion of sperm with abnormal [TB dark cells (TBDCs)] and normal [TB light cells (TBLCs)] chromatin structure were set by the ROC curve analysis. RESULTS: Thresholds of 45% TBDC and 20% TBLC were highly predictive for infertility (specificity of the test: 92 and 90%, respectively), but they were poor predictors of the fertility (sensitivity of the test: 42 and 32%, respectively). Odds ratio for infertility was 7.5 [95% confidence interval (CI): 2.7-20.8] when the 45% TBDC threshold was used and 4.4 (95% CI: 1.7-11.6) when the 20% TBLC threshold was used. CONCLUSIONS: The TB test can be suggested for clinical use as a complementary test for standard semen analysis to diagnose male infertility. PMID- 19304994 TI - Validation of data and indicators in the Danish Cholecystectomy Database. AB - OBJECTIVE: In The Danish Cholecystectomy Database (DCD), quality indicators are derived from clinical data in combination with administrative data from the National Patient Registry. The indicators 'Length of postoperative stay < or =1 day and no readmission', 'Length of stay (LOS) >3 days and/or readmission', 'Additional procedures within 30 days', 'Reconstructive bile duct surgery', 'Other surgery of the bile duct' and 'Death within 30 days' are all derived from administrative data. This study investigates the validity of the administrative data and evaluates the association between these indicators and postoperative complications. RESEARCH DESIGN AND SUBJECTS: Data from 1360 medical records of patients undergoing cholecystectomy were compared with the relevant administrative data from the National Patient Registry. The medical records served as the 'gold standard'. The association between the individual indicators and the occurrence of a postoperative complication was assessed. MEASURES: Validation of administrative data against the gold standard was done by the calculation of per cent agreement (including kappa-values) sensitivity/specificity and predictive values. The association between indicators and complications was analysed with crude event rates and odds ratios. RESULTS: The validity of the administrative data was excellent (97.1-100% agreement, kappa = 0.73-1.00). All of the indicators except 'Other bile duct surgery' were significantly associated with postoperative complications. A subdivision of some indicators strengthened the associations. CONCLUSIONS: The DCD is a valid method for monitoring the quality of cholecystectomy in Denmark. PMID- 19304995 TI - Priming and re-drying improve the survival of mature seeds of Digitalis purpurea during storage. AB - BACKGROUND AND AIMS: Most priming studies have been conducted on commercial seed lots of unspecified uniformity and maturity, and subsequent seed longevity has been reported to both increase and decrease. Here a seed lot of Digitalis purpurea L. with relatively uniform maturity and known history was used to analyse the effects of priming on seed longevity in air-dry storage. METHODS: Seeds collected close to natural dispersal and dried at 15% relative humidity (RH), 15 degrees C, were placed into experimental storage (60% RH, 45 degrees C) for 14 or 28 d, primed for 48 h at 0, -1, -2, -5, -10 or -15 MPa, re-equilibrated (47% RH, 20 degrees C) and then returned to storage. Further seed samples were primed for 2 or 48 h at -1 MPa and either dried at 15% RH, 15 degrees C or immediately re-equilibrated for experimental storage. Finally, some seeds were given up to three cycles of experimental storage and priming (48 h at -1 MPa). KEY RESULTS: Priming at -1 MPa had a variable effect on subsequent survival during experimental storage. The shortest lived seeds in the control population showed slightly increased life spans; the longer lived seeds showed reduced life spans. In contrast, seeds first stored for 14 or 28 d before priming had substantially increased life spans. The increase tended to be greatest in the shortest lived fraction of the seed population. Both the period of rehydration and the subsequent drying conditions had significant effects on longevity. Interrupting air-dry storage with additional cycles of priming also increased longevity. CONCLUSIONS: The extent of prior deterioration and the post-priming desiccation environment affect the benefits of priming to the subsequent survival of mature seeds. Rehydration-dehydration treatments may have potential as an adjunct or alternative to the regeneration of seed accessions maintained in gene banks for plant biodiversity conservation or plant breeding. PMID- 19304996 TI - Uniting pattern and process in plant-animal mutualistic networks: a review. AB - BACKGROUND: Ecologists and evolutionary biologists are becoming increasingly interested in networks as a framework to study plant-animal mutualisms within their ecological context. Although such focus on networks has brought about important insights into the structure of these interactions, relatively little is still known about the mechanisms behind these patterns. SCOPE: The aim in this paper is to offer an overview of the mechanisms influencing the structure of plant-animal mutualistic networks. A brief summary is presented of the salient network patterns, the potential mechanisms are discussed and the studies that have evaluated them are reviewed. This review shows that researchers of plant animal mutualisms have made substantial progress in the understanding of the processes behind the patterns observed in mutualistic networks. At the same time, we are still far from a thorough, integrative mechanistic understanding. We close with specific suggestions for directions of future research, which include developing methods to evaluate the relative importance of mechanisms influencing network patterns and focusing research efforts on selected representative study systems throughout the world. PMID- 19304997 TI - The molecular biology of seasonal flowering-responses in Arabidopsis and the cereals. AB - BACKGROUND: In arabidopsis (Arabidopsis thaliana), FLOWERING LOCUS T (FT) and FLOWERING LOCUS C (FLC) play key roles in regulating seasonal flowering-responses to synchronize flowering with optimal conditions. FT is a promoter of flowering activated by long days and by warm conditions. FLC represses FT to delay flowering until plants experience winter. SCOPE: The identification of genes controlling flowering in cereals allows comparison of the molecular pathways controlling seasonal flowering-responses in cereals with those of arabidopsis. The role of FT has been conserved between arabidopsis and cereals; FT-like genes trigger flowering in response to short days in rice or long days in temperate cereals, such as wheat (Triticum aestivum) and barley (Hordeum vulgare). Many varieties of wheat and barley require vernalization to flower but FLC-like genes have not been identified in cereals. Instead, VERNALIZATION2 (VRN2) inhibits long day induction of FT-like1 (FT1) prior to winter. VERNALIZATION1 (VRN1) is activated by low-temperatures during winter to repress VRN2 and to allow the long day response to occur in spring. In rice (Oryza sativa) a VRN2-like gene Ghd7, which influences grain number, plant height and heading date, represses the FT like gene Heading date 3a (Hd3a) in long days, suggesting a broader role for VRN2 like genes in regulating day-length responses in cereals. Other genes, including Early heading date (Ehd1), Oryza sativa MADS51 (OsMADS51) and INDETERMINATE1 (OsID1) up-regulate Hd3a in short days. These genes might account for the different day-length response of rice compared with the temperate cereals. No genes homologous to VRN2, Ehd1, Ehd2 or OsMADS51 occur in arabidopsis. CONCLUSIONS: It seems that different genes regulate FT orthologues to elicit seasonal flowering-responses in arabidopsis and the cereals. This highlights the need for more detailed study into the molecular basis of seasonal flowering responses in cereal crops or in closely related model plants such as Brachypodium distachyon. PMID- 19304998 TI - A conserved 3' extension in unusual group II introns is important for efficient second-step splicing. AB - The B.c.I4 group II intron from Bacillus cereus ATCC 10987 harbors an unusual 3' extension. Here, we report the discovery of four additional group II introns with a similar 3' extension in Bacillus thuringiensis kurstaki 4D1 that splice at analogous positions 53/56 nt downstream of domain VI in vivo. Phylogenetic analyses revealed that the introns are only 47-61% identical to each other. Strikingly, they do not form a single evolutionary lineage even though they belong to the same Bacterial B class. The extension of these introns is predicted to form a conserved two-stem-loop structure. Mutational analysis in vitro showed that the smaller stem S1 is not critical for self-splicing, whereas the larger stem S2 is important for efficient exon ligation and lariat release in presence of the extension. This study clearly demonstrates that previously reported B.c.I4 is not a single example of a specialized intron, but forms a new functional class with an unusual mode that ensures proper positioning of the 3' splice site. PMID- 19304999 TI - Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells. AB - The envelope glycoprotein of human immunodeficiency virus (HIV) consists of an exterior glycoprotein (gp120) and a trans-membrane domain (gp41) and has an important role in viral entry into cells. HIV-1 entry has been validated as a clinically relevant anti-viral strategy for drug discovery. In the present work, several 2'-F substituted RNA aptamers that bind to the HIV-1(BaL) gp120 protein with nanomole affinity were isolated from a RNA library by the SELEX (Systematic Evolution of Ligands by EXponential enrichment) procedure. From two of these aptamers we created a series of new dual inhibitory function anti-gp120 aptamer siRNA chimeras. The aptamers and aptamer-siRNA chimeras specifically bind to and are internalized into cells expressing HIV gp160. The Dicer-substrate siRNA delivered by the aptamers is functionally processed by Dicer, resulting in specific inhibition of HIV-1 replication and infectivity in cultured CEM T-cells and primary blood mononuclear cells (PBMCs). Moreover, we have introduced a 'sticky' sequence onto a chemically synthesized aptamer which facilitates attachment of the Dicer substrate siRNAs for potential multiplexing. Our results provide a set of novel inhibitory agents for blocking HIV replication and further validate the use of aptamers for delivery of Dicer substrate siRNAs. PMID- 19305000 TI - Genome sequence comparison of Col and Ler lines reveals the dynamic nature of Arabidopsis chromosomes. AB - Large differences in plant genome sizes are mainly due to numerous events of insertions or deletions (indels). The balance between these events determines the evolutionary direction of genome changes. To address the question of what phenomena trigger these alterations, we compared the genomic sequences of two Arabidopsis thaliana lines, Columbia (Col) and Landsberg erecta (Ler). Based on the resulting alignments large indels (>100 bp) within these two genomes were analysed. There are approximately 8500 large indels accounting for the differences between the two genomes. The genetic basis of their origin was distinguished as three main categories: unequal recombination (Urec)-derived, illegitimate recombination (Illrec)-derived and transposable elements (TE) derived. A detailed study of their distribution and size variation along chromosomes, together with a correlation analyses, allowed us to demonstrate the impact of particular recombination-based mechanisms on the plant genome evolution. The results show that unequal recombination is not efficient in the removal of TEs within the pericentromeric regions. Moreover, we discovered an unexpectedly high influence of large indels on gene evolution pointing out significant differences between the various gene families. For the first time, we present convincing evidence that somatic events do play an important role in plant genome evolution. PMID- 19305001 TI - XRCC1 interacts with the p58 subunit of DNA Pol alpha-primase and may coordinate DNA repair and replication during S phase. AB - Repair of single-stranded DNA breaks before DNA replication is critical in maintaining genomic stability; however, how cells deal with these lesions during S phase is not clear. Using combined approaches of proteomics and in vitro and in vivo protein-protein interaction, we identified the p58 subunit of DNA Pol alpha primase as a new binding partner of XRCC1, a key protein of the single strand break repair (SSBR) complex. In vitro experiments reveal that the binding of poly(ADP-ribose) to p58 inhibits primase activity by competition with its DNA binding property. Overexpression of the XRCC1-BRCT1 domain in HeLa cells induces poly(ADP-ribose) synthesis, PARP-1 and XRCC1-BRCT1 poly(ADP-ribosyl)ation and a strong S phase delay in the presence of DNA damage. Addition of recombinant XRCC1 BRCT1 to Xenopus egg extracts slows down DNA synthesis and inhibits the binding of PCNA, but not MCM2 to alkylated chromatin, thus indicating interference with the assembly of functional replication forks. Altogether these results suggest a critical role for XRCC1 in connecting the SSBR machinery with the replication fork to halt DNA synthesis in response to DNA damage. PMID- 19305005 TI - International disease surveillance. International disease monitoring, October to December 2008. PMID- 19305006 TI - Analgesia for cats after ovariohysterectomy with either buprenorphine or carprofen alone or in combination. AB - Eighty-four female cats undergoing ovariohysterectomy in a blinded, randomised, prospective clinical study were assigned to one of three groups of 28 to receive either 0.01 mg/kg buprenorphine (group B), 4 mg/kg carprofen (group C), or the same doses of both drugs (group BC). A dynamic and interactive visual analogue scale (DIVAS) from 0 to 100 mm, and a simple descriptive scale (SDS) from 0 to 4 were used to evaluate the cats' degree of analgesia and sedation for 24 hours postoperatively. There was no significant difference in the cats' sedation scores by SDS or DIVAS, and no difference in their pain scores by DIVAS. By SDS, the cats in group BC had significantly lower pain scores than the cats in group C (P<0.001) and group B (P<0.05). Nine of the cats in group B, nine in group C and five in group BC required rescue analgesia, and the cats in group C required rescue earlier than those in group B (P<0.05). PMID- 19305007 TI - Pilot study of the effect of individualised homeopathy on the pruritus associated with atopic dermatitis in dogs. AB - Twenty dogs with confirmed atopic dermatitis were treated with homeopathy. In the first phase of this pilot study, all of the dogs were treated by a veterinary homeopath with individualised remedies prescribed on the basis of the dog's cutaneous signs and constitutional characteristics. The response to treatment was assessed by scoring the severity of pruritus from 0 to 10 on a validated scale. The dogs were evaluated at monthly intervals for at least two months. In 15 cases, the owners reported no improvement following homeopathic treatment. In the other five cases, the owners believed that the homeopathic treatment was associated with a substantial improvement, and reported reductions in pruritus scores ranging from 64 to 100 per cent. These five dogs were selected for the second phase of the study, in which homeopathic remedies were tested against placebos in a randomised and blinded trial. In one of these dogs, atopic dermatitis resolved completely and so this dog could not participate in phase 2; another dog was euthanased because of status epilepticus before phase 2 could be started. In the remaining three cases, the owners correctly distinguished between the placebo and homeopathic remedies, and reported reductions in the pruritus score of 0, 0.2 and 0.8 following placebo treatment and 4.3, 2.4 and 3.0, respectively, following the remedy. PMID- 19305008 TI - Preliminary epidemiological analysis of BSE cases born after 2000 in Great Britain. PMID- 19305009 TI - Metastasising liposarcoma of bone in a young dog. PMID- 19305010 TI - Diagnosis by PCR of Coxiella burnetii in aborted fetuses of domestic ruminants in Portugal. PMID- 19305012 TI - Register of premises supplying veterinary medicines. PMID- 19305013 TI - What is veterinary forensics? PMID- 19305014 TI - Retinal pigment epithelial dystrophy in the Briard. PMID- 19305015 TI - Spironucleosis in gamebirds. PMID- 19305016 TI - Changes to package labelling of Propalin. PMID- 19305017 TI - Supply of Deccox 25 Individual Treatment Packs. PMID- 19305018 TI - Septic pneumonia and pyothorax in dogs. PMID- 19305019 TI - Separate elements within a single IQ-like motif in adenylyl cyclase type 8 impart ca2+/calmodulin binding and autoinhibition. AB - The ubiquitous Ca(2+)-sensing protein calmodulin (CaM) fulfills its numerous signaling functions through a wide range of modular binding and activation mechanisms. By activating adenylyl cyclases (ACs) 1 and 8, Ca(2+) acting via calmodulin impacts on the signaling of the other major cellular second messenger cAMP. In possessing two CaM-binding domains, a 1-5-8-14 motif at the N terminus and an IQ-like motif (IQlm) at the C terminus, AC8 offers particularly sophisticated regulatory possibilities. The IQlm has remained unexplored beyond the suggestion that it bound CaM, and the larger C2b region of which it is part was involved in the relief of autoinhibition of AC8. Here we attempt to distinguish the function of individual residues of the IQlm. From a complementary approach of in vitro and cell population AC activity assays, as well as CaM binding, we propose that the IQlm alone, and not the majority of the C2b, imparts CaM binding and autoinhibitory functions. Moreover, this duality of function is spatially separated and depends on amino acid side-chain character. Accordingly, residues critical for CaM binding are positively charged and clustered toward the C terminus, and those essential for the maintenance of autoinhibition are hydrophobic and more N-terminal. Secondary structure prediction of the IQlm supports this separation, with an ideally placed break in the alpha-helical nature of the sequence. We additionally find that the N and C termini of AC8 interact, which is an association specifically abrogated by fully Ca(2+)-bound, but not Ca(2+)-free, CaM. These data support a sophisticated activation mechanism of AC8 by CaM, in which the duality of the IQlm function is critical. PMID- 19305021 TI - Importance of mesophyll diffusion conductance in estimation of plant photosynthesis in the field. AB - Mesophyll diffusion conductance to CO(2) (g(m)) is an important leaf characteristic determining the drawdown of CO(2) from substomatal cavities (C(i)) to chloroplasts (C(C)). Finite g(m) results in modifications in the shape of the net assimilation (A) versus C(i) response curves, with the final outcome of reduced maximal carboxylase activity of Rubisco (V(cmax)), and a greater ratio of the capacity for photosynthetic electron transport to V(cmax) (J(max)/V(cmax)) and alterations in mitochondrial respiration rate (R(d)) when estimated from A/C(i) responses without considering g(m). The influence of different Farquhar et al. model parameterizations on daily photosynthesis under non-stressed (C(i) kept constant throughout the day) and stressed conditions (mid-day reduction in C(i)) was compared. The model was parameterized on the basis of A/C(C) curves and A/C(i) curves using both the conventional fitting procedure (V(cmax) and R(d) fitted separately to the linear part of the response curve and J(max) to the saturating part) and a procedure that fitted all parameters simultaneously. The analyses demonstrated that A/C(i) parameterizations overestimated daily assimilation by 6-8% for high g(m) values, while they underestimated if by up to 70% for low g(m) values. Qualitative differences between the A/C(i) and A/C(C) parameterizations were observed under stressed conditions, when underestimated V(cmax) and overestimated R(d) of A/C(i) parameterizations resulted in excessive mid-day depression of photosynthesis. Comparison with measured diurnal assimilation rates in the Mediterranean sclerophyll species Quercus ilex under drought further supported this bias of A/C(i) parameterizations. While A/C(i) parameterization predicted negative carbon balance at mid-day, actual measurements and simulations with the A/C(C) approach yielded positive carbon gain under these conditions. In addition, overall variation captured by the best A/C(i) parameterization was poor compared with the A/C(C) approach. This analysis strongly suggests that for correct parameterization of daily time-courses of photosynthesis under realistic field conditions, g(m) must be included in photosynthesis models. PMID- 19305020 TI - Effects of ezetimibe and simvastatin on apolipoprotein B metabolism in males with mixed hyperlipidemia. AB - Sixteen hyperlipidemic men were enrolled in a randomized, placebo-controlled, double-blind, cross-over study to evaluate the effect of ezetimibe 10 mg and simvastatin 40 mg, coadministered and alone, on the in vivo kinetics of apolipoprotein (apo) B-48 and B-100 in humans. Subjects underwent a primed constant infusion of a stable isotope in the fed state. The coadministration of simvastatin and ezetimibe significantly reduced plasma concentrations of cholesterol (-43.0%), LDL-C (-53.6%), and triglycerides (-44.0%). Triglyceride rich lipoproteins (TRL) apoB-48 pool size (PS) was significantly decreased ( 48.9%) following combination therapy mainly through a significant reduction in TRL apoB-48 production rate (PR) (-38.0%). The fractional catabolic rate (FCR) of VLDL and LDL apoB-100 were significantly increased with all treatment modalities compared with placebo, leading to a significant reduction in the PS of these fractions. We also observed a positive correlation between changes in TRL apoB-48 PS and changes in TRL apoB-48 PR (r = 0.85; P < 0.0001) with combination therapy. Our results indicate that treatment with simvastatin plus ezetimibe is effective in reducing plasma TRL apoB-48 levels and that this effect is most likely mediated by a reduction in the intestinal secretion of TRL apoB-48. Our study also indicated that the reduction in LDL-C concentration following combination therapy is mainly driven by an increase in FCR of apoB-100 containing lipoproteins. PMID- 19305022 TI - Expression of the SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE1 (SERK1) gene is associated with developmental change in the life cycle of the model legume Medicago truncatula. AB - SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) genes have been demonstrated to play a role in somatic embryogenesis in several plant species. As more is learnt about these genes, the view of their role in plant development has broadened. The Medicago truncatula MtSERK1 gene has been associated with somatic embryogenesis and in vitro root formation. In order to study the role of MtSERK1 in development further, the MtSERK1 promoter sequence has been isolated and cloned into a promoter-GUS analysis vector. SERK1 promoter-driven GUS expression was studied in A. tumefaciens-transformed cultures and regenerated plants, in A. rhizogenes transformed root clones, and in nodulation. In embryogenic cultures, GUS staining is detected after 2 d of culture at the edge of the explant and around vascular tissue. Expression at the explant edge intensifies over subsequent days and then is lost from the edge as callus formation moves inward. MtSERK1 expression appears to be associated with new callus formation. When somatic embryos form, GUS staining occurs throughout embryo development. Zygotic embryos show expression until the heart stage. The in planta studies reveal a number of interesting expression patterns. There appear to be three types. (i) Expression associated with the primary meristems of the root and shoot and the newly formed meristems of the lateral roots and nodule. (ii) Expression at the junction between one type of tissue or organ and another. (iii) Expression associated with the vascular tissue procambial cells. The data led us to conclude that MtSERK1 expression is associated with developmental change, possibly reflecting cellular reprogramming. PMID- 19305023 TI - [Genetic background of isolated forms of congenital malformations of the hand]. AB - Congenital hand malformations comprise a clinically and etiologically heterogeneous group of developmental anomalies. They may occur as an isolated trait or be a part of a syndrome. In recent years, enormous progress in revealing the molecular background of congenital hand anomalies has been made. It proved helpful for better understanding of function of the genes responsible for embroynic limb development in humans. This review presents a classification of hand malformations, which takes into account not only differences of anatomic and morphological origin, but also distinct genetic background. Since there is an abundance of syndromes with congenital hand anomalies, the paper is focused mainly on the non-syndromic hand malformations and their genetic etiology. PMID- 19305024 TI - [Genetically determined human susceptibility to selected infectious diseases]. AB - As predictions show infectious diseases were, are and will be, responsible for a significant percentage (more than 12% in the year 2030) of deaths worldwide. Infectious diseases are, according to J.B.S. Haldane's theory, the major agent determining natural selection, as they lead to elimination of more susceptible people and only leave to survive these, who are more resistant. It has been revealed that susceptibility to pathogens varies among ethnic groups. Explanation of this phenomenon can be found in the human genome. Standard genetic analysis led to identification of several gene variants which modulate susceptibility to particular infectious disease as well as its progression. HLA genes encoding major histocompatibility complex are one of the most interesting ones as they are reported to influence the susceptibility to a wide range of pathogens. It is also proved that in several cases many other genes take part in modulation of clinical outcome of the diseases. Alleles conferring partial or total protection against disease development have already been identified. This review presents results of selected research concerning genetically determined susceptibility to malaria, cholera, leprosy and HIV. PMID- 19305025 TI - [Variable clinical expression of familial Incontinentia Pigmenti syndrome - presentation of three cases]. AB - Incontinentia Pigmenti (IP, Bloch-Sulzberger syndrome, OMIM 308300) is a rare X linked dominant genodermatosis, usually lethal in males in the prenatal period. Wide spectrum of clinical expression consists of skin hyperpigmented lines and swirling patterns, dysplastic teeth and nails, and in 30% central nervous system abnormalities including seizures, microcephaly and intellectual disability (10% of cases). In 80% of IP cases, the disease is caused by a large-scale deletion of exons 4 to 10 of the NEMO gene. Three cases of variable expression of Incontinentia Pigmenti are presented. In a one-year-old girl, her mother and grandmother molecular analysis revealed the same typical deletion of the NEMO gene. In the proband, characteristic skin lesions were detected located over the trunk and lower limbs. Characteristic evolution of the changes was observed. In the mother, expression of the disease was much milder, whereas in the grandmother lesions were restricted to the fingernails. Clinical characteristics and pedigree data are described. PMID- 19305026 TI - [Parenteral symptoms and intestinal complications in children with inflammatory bowel diseases in relation to card15 mutation]. AB - THE AIM OF THE STUDY: Was evaluation of the incidence of parenteral symptoms and complications in children with inflammatory bowel disease and their analysis in relation to the examined mutations of CARD15 gene. PATIENTS AND METHODS: The study involved 38 children with Crohn's disease, aged from 5 to18 years (median14) and 40 children with ulcerative colitis, aged from 6 to18 years (median14). The control group included 23 children, aged from 4 to 18 years (median15), with functional disorders of the alimentary tract resulting from lactose intolerance. In all the examined patients as well as in the control group, mutations R702W, G908R and L1007fs of the CARD15 gene were determined, according to the protocol described by Tukel et al. RESULTS: Parenteral symptoms, in the group of children with Crohn's disease, manifested as arthritis and erythema nodosum, were observed in 7 patients (18.4%), whereas in the group with ulcerative colitis they presented - in 4 children (10%). Intestinal complications in the form of stenosis, fistula, abscess and gastrointestinal bleeding were the most frequently observed changes in children with Crohn's disease (n=15; 39,5%). Parenteral symptoms were statistically significantly more frequent in children with Crohn's disease and with at least one mutation of CARD15 gene. Intestinal complications statistically appeared more often in children with Crohn's disease and mutation L1007fs. CONCLUSIONS: 1. Parenteral symptoms and intestinal complications occurred more frequently in the group of children with Crohn's disease, in comparison with the children with ulcerative colitis. 2. We observed a relation between parenteral symptoms and at least one mutation of CARD15 gene and a relation between intestinal complications and L1007fs mutation. PMID- 19305027 TI - [Evaluation of selected endocrine complications in patients treated with auto- and allo-haematopoietic stem cell transplantation]. AB - AIM: The aim of this study was to evaluate the endocrine complications, in particular disorders of growth and thyroid function and glucose metabolism dysfunctions in patients treated with allo- and auto-haematopoietic stem cell transplantation (HSCT). MATERIAL AND METHODS: The investigated group consisted of: I. 16 patients after auto-HSCT (6 girls, 10 boys) aged 3-20 years (average 10,8+/-) because of acute myelogenous leukaemia (n=5), non Hodgkin lymphoma (n=3), neuroblastoma (n=3), embryonal cancer (n=2), medulloblastoma (n=1), Ewing's sarcoma/PNET (n=1), hyper eosinophilic syndrome (n=1). High dose chemiotherapy (HDC/T) included: BU/MEL (busulfan/melfalan) (n=7), BEAM (carmustine, eteposide, cytosine arabinose, melfalan) (n=3). II. 30 patients after allo-HSCT (20 girls, 10 boys) aged 3-17 years (average 9,56). Indication for HSCT was acute lymphoblastic leukaemia (n=11), acute myelogenous leukaemia (n=5), chronic myeloid leukaemia-CML (n=6), myelodysplastic syndromes (n=2), non Hodgkin lymphoma (n=1), juvenile myelomonocytic leukemia (n=1), severe aplastic anaemia (n=1), Blackfan-Diamond anaemia (n=1), severe combined immune deficiency (n=1), rhabdomyosarcoma (n=1). The patients underwent the following types of transplantation: HSCT of matched sibling donor (n=13), HSCT of matched unrelated donor (n=11) and HLA-mismatched related donor (n=6). The preparative regimens consisted of HDC/T usually BU/MEL (n=3); BU/CY/VP (busulfan, cyclophosphamide, etoposide) (6); BU/CY/ATG (anti-thymocyte globulin) (n=5), VP/ATG/TBI (total body irradiation) (n=3). 19 children received CI (cranial irradiation) prior to grafting: auto-HSCT (n=6) and allo-HSCT (n=13) and 6 patients underwent TBI. 18 children received high steroid doses at least 28 days before transplant, 4 patients in the auto-HSCT group, and in the allo-HSCT group 14 patients before and 20 after HSCT procedure. The analysis of thyroid-stimulating hormone (TSH), free triiodothyronine (fT3), free thyroxine (fT4), insulin-like growth factor 1 (IGF-1), insulin-like growth factor binding protein 3 (IGFBP-3), hemoglobin A1c (HbA1c), prolactine (PRL), oral glucose tolerance test, growth hormone (GH test) and thyrotropin releasing hormone (TRH) test was performed in each case. RESULTS: Hypothyroidism was found in 5 patients (3 after allo-HSCT, 2 after auto-HSCT). Thyroid hormone substitution was applied. No case of hyperthyroidism was diagnosed. Growth deficit was found in 8 patients (6 girls, 2 boys) between 13 to 70 months after allo-transplantation (average 36 months). Three children from the above group received CI. Growth hormone substitution was applied in 1 girl (ALL, HLA MM REL, CI). An impaired excretion of GH after stimulation was diagnosed in 14 pts (10 after allo-HSCT, 4 after auto-HSCT). The growth process should still be observed in this subgroup. Glucose intolerance was found in 7 patients: in 4 treated with auto-HSCT and in 3 after allo-HSCT. Diabetes mellitus was diagnosed in none of them. An impaired glucose tolerance curve with increased excretion of insulin was diagnosed in 12 children. CONCLUSIONS: Early endocrinological care is necessary in patients treated both with auto-HSCT and allo-HSCT due to high risk of hormonal disorders. PMID- 19305028 TI - [Neutropenia in infancy - sometimes chronic and benign - own experiences]. AB - Chronic neutropenia is a decrease in circulating neutrophils in the peripheral blood lasting over 6 months. Values need to be refered with the age and race. In children aged 2 weeks to 12 months reffered values are above 1000/03BCL. There are congenital and aquired reasons of neutropenia in infancy. The most common type of chronic neutropenia in infants is chronic, benign neutropenia (AIN). Authors present ten infants between three and six months with chronic, benign neutropenia. The reason of ordering laboratory tests at outpatient clinic were benign upper respiratory tract infections (four cases), pallor (four cases) and on parental demand (one case). In one infant neutropenia was observed during treatment of pneumonia at a district hospital. PMID- 19305029 TI - [Risk factors of surgical site infection in newborn infants]. AB - AIM OF THE STUDY: Was to estimate the risk factors for surgical site infections (SSJ) in the newborn infants. MATERIALS AND METHODS: We retrospectively analyzed medical records of 381 operated newborn infants in order to estimate the rate of SSI and risk factors. All types of operations were divided according to contamination of surgical wound /CDC classification: clean, clean-contaminated, contaminated and dirty infected/. Additionally we evaluated risk factors such as: birth weight, gestational age, congenital infection, additional surgery, other congenital defect and invasive procedures like: mechanical ventilation and presence of central venous line. Descriptive statistics and odds ratios with 95% confidence intervals were applied in univariate statistical analysis. RESULTS: The mean incidence of SSI was 37%. According to CDC classification SSI rate were 27%, 35%, 46%, 71% respectively. The most important risk factors of SSI were: mechanical ventilation (IS:10.80), central line (IS:8.20), birth weight below 1500 g (IS:5.03) and congenital infection (IS:4.74). The risk of SSI depended on the type of surgery. The incidence of infections was significantly higher for contaminated and dirty-infected wounds than for clean and clean-contaminated. Risk factors for SSI were similar for clean and clean-contaminated wounds as for all study group. CONCLUSIONS: The premature newborns who underwent surgery, were mechanically ventilated, with venous access had the highest risk of SSI. The significance of risk factors was the same for the total study group for every type of surgery wound. PMID- 19305030 TI - [Transposition of great arteries in a newborn with congenital diaphragmatic hernia - case report]. AB - A female newborn was delivered by Caesarean section in 36/37 week of pregnancy, due to prenatally diagnosed congenital diaphragmatic hernia. She was admitted to the intensive care unit in the first minutes of life. It was the first uneventful pregnancy of a 25 year old woman. The newborn's Apgar score was 0, at both 1(st) and 5(th) minute of life. She was immediately intubated, given adrenaline and 4.2% NaHCO(3). High frequency oscillatory ventilation, with high ventilation parameters was initiated. Right-sided pneumothorax was found by auscultation and pleural drainage was installed. On X-ray a left-sided congenital diaphragmatic hernia was noted with presence of the stomach, spleen, small intestine and left lobe of the liver in the thorax. Because of deteriorating clinical condition fluid resuscitation and continuous noradrenaline infusion were used. Despite implemented treatment, severe bradycardia and hypotension developed. Patient died 1.5 hours after birth. On autopsy left-sided diaphragmatic hernia, bilateral pneumothorax, pneumoperitoneum, severe lung hypoplasia and transposition of the great arteries were found. Prenatal diagnosis of cardiac defects by routine ultrasonography may be difficult in a neonate with diaphragmatic hernia, due to altered anatomical conditions. Postnatal echocardiographic examination of the heart is suggested in such infants to exclude the presence of cardiovascular anomalies. PMID- 19305031 TI - [Tolerance of two lipid emulsions used in parenterally-fed premature infants - a comparative study]. AB - INTRODUCTION: Parenteral nutrition is essential for premature infants during their first days of extrauterine life, when enteral feeding is not tolerated. Lipid emulsions are an integral part of parenteral nutrition. The aim of the study was to compare the effects of two lipid emulsions, based on olive-oil and soybean oil, used in parenteral nutrition of premature infants on: plasma lipid concentrations and hyperbilirubinemia based on plasma bilirubin levels and phototherapy times. MATERIAL AND METHODS: Forty-four premature infants aged less than 32 weeks of gestation and with birth weight below 1500 g were randomized within first 48 hours of life, to receive in double-blind manner olive oil based (ClinOleic) or soybean oil based (Ivelip) emulsions. Plasma lipid concentration and bilirubin level were determined at 7(th) day of life. RESULTS: Thirty-eight infants completed the study, 18 in the olive oil group (27+/-2 GW, 936+/-218 g) and 20 in the soybean oil group (27+/-2 GW, 924+/-221 g). Both lipid emulsions were well tolerated and plasma lipid concentration were within a normal range, however in olive-oil group plasma cholesterol concentration (159+/-19.89 mg% vs. 128+/-8.85 mg%, p<0.0126) and plasma LDL concentration (103.36+/-8.85 mg% vs. 78.87+/-7.59 mg%, p<0.0342) were significantly higher. Plasma bilirubin levels and phototherapy times did not differ among the groups. CONCLUSIONS: Both lipid emulsions were well tolerated, maintained normal plasma lipid concentration and did not increase plasma bilirubin level. PMID- 19305032 TI - [Etiological agents of urinary tract infections in children treated at the Institute of Mother and Child]. AB - BACKGROUND: Urinary tract is a common site of bacterial infections in children. The main etiological agents of infection are Gram negative bacteria from the Enterobacteriaceae in particular of intestinal origin. The uropathogenic strains may indicate resistance for many antibacterial drugs. AIM: The aim of this study was an analysis of urine samples from outpatient or hospitalized children treated at the Institute of Mother and Child in Warsaw, in the years 2006-2007. The frequency of individual bacterial species and antibiotic susceptibility were investigated. MATERIALS AND METHODS: In all, 2096 urine samples from patients treated at the Institute of Mother and Child were investigated. Bacteriuria was examined and antibiotics susceptibility was estimated. Biochemical tests were used for the identification of uropathogenic strains. Antibiotic susceptibility was examined using standard diffusion discs methods. RESULTS: During 2006-2007, in all 2096 urine samples were taken, of which 313 were positive (14.9%). E. coli was the most often isolated bacteria (54%) and enterococci dominated in Gram positive bacteria. CONCLUSIONS: Enterobacteriaceae were the most frequently isolated bacteria. Most of the antibiotics used in the treatment of urinary tract infections had high antibacterial activity except for ampicillin. PMID- 19305033 TI - [Congenital cystic adenomatoid lung malformation - diagnosis and treatment - based on literature and own experiences]. AB - Congenital cystic adenomatoid malformation (CCAM) is an uncommon congenital abnormality of the lung which has a wide spectrum of potential outcomes, ranging from hydrops and severe respiratory distress with pulmonary hypoplasia, to resolution of the lesion either antenatally or postnatally. Most of the babies are asymtomaptic. It is caused by arrest of normal foetal pulmonary maturation. The prognosis is generally good. In cases where the lesion persists, surgery is recommended. CCAM is an important diagnosis and can be suspected on routine antenatal ultrasound. It has implication for both the ongoing pregnancy, at delivery and later in life. The authors present a male newborn with CCAM diagnosed during the neonatal period. The diagnosis was based on CT scan and histopathological examination. Characteristic plural cystic defects in the lungs were found. The patient was referred for surgical treatment. PMID- 19305034 TI - [Stevens-Johnson syndrome in the literature and authors' own studies]. AB - Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis (Lyell Syndrome) are severe, multisystem disease caused mainly by reaction to drugs. The clinical features include: changes on the skin and mucosa as well as lesions in the internal organs. There are no standards of treatment in this diesease group. There are ongoing trials in various groups of drugs. Apart from providing the appriopriate conditions and symptomatic treatment, immunoglobulins, cytostatics and plasmapheresis are used. The authors present the clinical course of a patient with Stevens-Johnsons Syndrome, probably caused by klaritromycin. The 13 y.o. boy was admitted to a Department of Paediatric Surgery specializing in treatment of burns. Immunoglobulin, cytostatics, antibacterial and antiviral drugs were used as well as topical medicines routinely applied in burns. The method proved to give good results. The treatment of Stevens-Johnsons Syndrome is difficult and expensive. Due to lack of standards, the treatment depends on one's own experience and up to date literature. PMID- 19305036 TI - The risk of developing a second cancer after receiving craniospinal proton irradiation. AB - The purpose of this work was to compare the risk of developing a second cancer after craniospinal irradiation using photon versus proton radiotherapy by means of simulation studies designed to account for the effects of neutron exposures. Craniospinal irradiation of a male phantom was calculated for passively-scattered and scanned-beam proton treatment units. Organ doses were estimated from treatment plans; for the proton treatments, the amount of stray radiation was calculated separately using the Monte Carlo method. The organ doses were converted to risk of cancer incidence using a standard formalism developed for radiation protection purposes. The total lifetime risk of second cancer due exclusively to stray radiation was 1.5% for the passively scattered treatment versus 0.8% for the scanned proton beam treatment. Taking into account the therapeutic and stray radiation fields, the risk of second cancer from intensity modulated radiation therapy and conventional radiotherapy photon treatments were 7 and 12 times higher than the risk associated with scanned-beam proton therapy, respectively, and 6 and 11 times higher than with passively scattered proton therapy, respectively. Simulations revealed that both passively scattered and scanned-beam proton therapies confer significantly lower risks of second cancers than 6 MV conventional and intensity-modulated photon therapies. PMID- 19305037 TI - The calibration of parallel-plate electron ionization chambers at NPL for use with the IPEM 2003 code of practice: summary data. AB - The most recent electron dosimetry code of practice for radiotherapy written by the Institute of Physics and Engineering in Medicine was published in 2003 and is based on the NPL electron absorbed dose to water calibration service. NPL has calibrated many Scanditronix type NACP-02 and PTW Roos type 34001 parallel plate ionization chambers in terms of absorbed dose to water, for use with the code of practice. The results of the calibrations of these chamber types summarized here include the absorbed dose to water sensitivity, where the mean calibration factor standard deviations are 5.8% for NACP-02 chambers and 1.1% for PTW Roos chambers. The correction for the polarity effect is shown to be small (less than 0.2% for all beam qualities) but with a discernible beam quality dependence. The correction for recombination is shown to be consistent and reproducible, and an analysis of these results suggests that the plate separation of the NACP-02 chambers is more variable from chamber to chamber than with the PTW Roos chambers. The calibration of these chambers is shown to be repeatable within +/ 0.2% over 2-3 years. It is also shown that check source measurements can be repeated within +/-0.3% over several years. The results justify the use of NACP 02 and PTW 34001 chambers as secondary standards, but also indicate that the PTW 34001 chambers show less variation from chamber to chamber. PMID- 19305039 TI - Letter to the Editor on 'Single-Arc IMRT?'. AB - In the note 'Single Arc IMRT?' (Bortfeld and Webb 2009 Phys. Med. Biol. 54 N9 20), Bortfeld and Webb present a theoretical investigation of static gantry IMRT (S-IMRT), single-arc IMRT and tomotherapy. Based on their assumptions they conclude that single-arc IMRT is inherently limited in treating complex cases without compromising delivery efficiency. Here we present an expansion of their work based on the capabilities of the Varian RapidArc single-arc IMRT system. Using the same theoretical framework we derive clinically deliverable single-arc IMRT plans based on these specific capabilities. In particular, we consider the range of leaf motion, the ability to rapidly and continuously vary the dose rate and the choice of collimator angle used for delivery. In contrast to the results of Bortfeld and Webb, our results show that single-arc IMRT plans can be generated that closely match the theoretical optimum. The disparity in the results of each investigation emphasizes that the capabilities of the delivery system, along with the ability of the optimization algorithm to exploit those capabilities, are of particular importance in single-arc IMRT. We conclude that, given the capabilities available with the RapidArc system, single-arc IMRT can produce complex treatment plans that are delivered efficiently (in approximately 2 min). PMID- 19305040 TI - The validation of tomotherapy dose calculations in low-density lung media. AB - The dose-calculation accuracy of the tomotherapy Hi-Art II(R) (Tomotherapy, Inc., Madison, WI) treatment planning system (TPS) in the presence of low-density lung media was investigated. In this evaluation, a custom-designed heterogeneous phantom mimicking the mediastinum geometry was used. Gammex LN300 and balsa wood were selected as two lung-equivalent materials with different densities. Film analysis and ionization chamber measurements were performed. Treatment plans for esophageal cancers were used in the evaluation. The agreement between the dose calculated by the TPS and the dose measured via ionization chambers was, in most cases, within 0.8%. Gamma analysis using 3% and 3 mm criteria for radiochromic film dosimetry showed that 98% and 95% of the measured dose distribution had passing gamma values < or =1 for LN300 and balsa wood, respectively. For a homogeneous water-equivalent phantom, 95% of the points passed the gamma test. It was found that for the interface between the low-density medium and water equivalent medium, the TPS calculated the dose distribution within acceptable limits. The phantom developed for this work enabled detailed quality-assurance testing under realistic conditions with heterogeneous media. PMID- 19305042 TI - Comments on 'Single-Arc IMRT?'. AB - We read with interest the article titled 'Single-Arc IMRT?' (Bortfeld and Webb 2009 Phys. Med. Biol. 54 N9-20) and feel it imperative to draw the attention of your readers to comments suggesting that the authors may not be fully aware of current developments in this field. As their paper was first submitted on 19th of August 2008, it could not have taken into account data presented at the AAPM, ESTRO and ASTRO meetings in 2008. In this letter, we would like to clarify some relevant aspects of RapidArc (Varian Medical Systems) as a modality for delivering single-arc treatment. PMID- 19305043 TI - Treatment planning and dose analysis for interstitial photodynamic therapy of prostate cancer. AB - With the development of new photosensitizers that are activated by light at longer wavelengths, interstitial photodynamic therapy (PDT) is emerging as a feasible alternative for the treatment of larger volumes of tissue. Described here is the application of PDT treatment planning software developed by our group to ensure complete coverage of larger, geometrically complex target volumes such as the prostate. In a phase II clinical trial of TOOKAD vascular targeted photodynamic therapy (VTP) for prostate cancer in patients who failed prior radiotherapy, the software was used to generate patient-specific treatment prescriptions for the number of treatment fibres, their lengths, their positions and the energy each delivered. The core of the software is a finite element solution to the light diffusion equation. Validation against in vivo light measurements indicated that the software could predict the location of an iso fluence contour to within approximately +/-2 mm. The same software was used to reconstruct the treatments that were actually delivered, thereby providing an analysis of the threshold light dose required for TOOKAD-VTP of the post irradiated prostate. The threshold light dose for VTP-induced prostate damage, as measured one week post-treatment using contrast-enhanced MRI, was found to be highly heterogeneous, both within and between patients. The minimum light dose received by 90% of the prostate, D(90), was determined from each patient's dose volume histogram and compared to six-month sextant biopsy results. No patient with a D(90) less than 23 J cm(-2) had complete biopsy response, while 8/13 (62%) of patients with a D(90) greater than 23 J cm(-2) had negative biopsies at six months. The doses received by the urethra and the rectal wall were also investigated. PMID- 19305044 TI - Initial evaluation of acoustic reflectors for the preservation of sensitive abdominal skin areas during MRgFUS treatment. AB - During MR-guided focused ultrasound (MRgFUS) treatments of uterine fibroids using ExAblate(R)2000 (InSightec, Haifa, Israel), individual tissue ablations are performed extracorporeally through the patient's abdomen using an annular array FUS transducer embedded within the MR table. Ultrasound intensities in the near field are below therapeutic levels and, under normal conditions, heating of the patient skin is minimal. However, increased absorption of ultrasound energy within sensitive skin areas or areas with differing acoustic properties, such as scars, may lead to skin burns and therefore these areas must be kept outside the near field of the FUS beam. Depending on their location and size the sensitive areas may either obstruct parts of the fibroid from being treated or prevent the entire MRgFUS treatment altogether. The purpose of this work is to evaluate acoustic reflector materials that can be applied to protect skin and the underlying sensitive areas. Reflection coefficients of cork (0.88) and foam (0.91) based materials were evaluated with a hydrophone. An ExAblate 2000 MRgFUS system was used to simulate clinical treatment with discs of reflector materials placed in a near field underneath a gel phantom. MR thermometry was used to monitor temperature elevations as well as the integrity of the focal spot. The phantom measurements showed acoustic shadow zones behind the reflectors with zone depths changing between 7 and 27 mm, for reflector disc diameters increasing from 10 to 30 mm (40 mm diameter discs completely blocked the FUS beam at the depth evaluated). The effects on thermal lesions due to the presence of the reflectors in the FUS beam were found to diminish with decreasing disc diameter and increasing sonication depth. For a 20 mm diameter disc and beyond 50 mm sonication depth, thermal lesions were minimally affected by the presence of the disc. No heating was observed on the skin side of the foam reflectors, as confirmed by measurements performed with adhesive temperature labels. We present these data and discuss possible applications to clinical MRgFUS treatments. PMID- 19305046 TI - Burden of invasive disease caused by Haemophilus influenzae type b in Asia. AB - Globally, Haemophilus influenzae type b (Hib) is one of the leading causes of childhood meningitis and pneumonia. Vaccines against Hib have been developed and introduced into routine immunization programs in most industrialized nations, in which the burden of Hib disease has been clearly demonstrated. In Asia, the burden of Hib disease has been perceived as relatively low compared with other parts of the world. However, the results of several recent studies have allowed for a more cautious assessment of the disease burden in different populations and settings throughout Asia. These studies suggest that the true burden of Hib disease may have been underestimated, and point to several key factors that may have contributed to this problem, such as antibiotic use prior to the analysis of blood and cerebrospinal fluid in infants and children suspected of having invasive Hib disease. Trials evaluating Hib vaccines in Asian populations have shown that they are effective and safe. These results may aid in the decision about whether to implement routine Hib vaccination in Asia and help to track the impact of Hib vaccination in the future. PMID- 19305045 TI - Stray radiation dose and second cancer risk for a pediatric patient receiving craniospinal irradiation with proton beams. AB - Proton beam radiotherapy unavoidably exposes healthy tissue to stray radiation emanating from the treatment unit and secondary radiation produced within the patient. These exposures provide no known benefit and may increase a patient's risk of developing a radiogenic cancer. The aims of this study were to calculate doses to major organs and tissues and to estimate second cancer risk from stray radiation following craniospinal irradiation (CSI) with proton therapy. This was accomplished using detailed Monte Carlo simulations of a passive-scattering proton treatment unit and a voxelized phantom to represent the patient. Equivalent doses, effective dose and corresponding risk for developing a fatal second cancer were calculated for a 10-year-old boy who received proton therapy. The proton treatment comprised CSI at 30.6 Gy plus a boost of 23.4 Gy to the clinical target volume. The predicted effective dose from stray radiation was 418 mSv, of which 344 mSv was from neutrons originating outside the patient; the remaining 74 mSv was caused by neutrons originating within the patient. This effective dose corresponds to an attributable lifetime risk of a fatal second cancer of 3.4%. The equivalent doses that predominated the effective dose from stray radiation were in the lungs, stomach and colon. These results establish a baseline estimate of the stray radiation dose and corresponding risk for a pediatric patient undergoing proton CSI and support the suitability of passively scattered proton beams for the treatment of central nervous system tumors in pediatric patients. PMID- 19305047 TI - Serodiversity and antimicrobial resistance pattern of Shigella isolates at Gondar University teaching hospital, Northwest Ethiopia. AB - The aims of this study were to determine the distribution of serogroups and serotypes as well as the antimicrobial resistance pattern of Shigella isolates from Gondar patients with acute diarrhea. A cross-sectional study was conducted from August 2006 to February 2008. Stool specimens were received from study subjects and cultured. Isolates were confirmed by biochemical and serological tests. The isolates were tested for antimicrobial susceptibility by the disc diffusion method. Of the 1,200 stool specimens, 90 (7.5%) yielded Shigella isolates with the serogroups of Shigella flexneri (72.2%), S. dysenteriae (10.0%), S. boydii (8.9%), and S. sonnei (8.9%). S. flexneri was the predominant serogroup. S. dysenteriae type 1 was absent, and S. sonnei was present. Eighty five (94.5%) of the isolates showed resistance to one or more drugs, and 79% of those were multi-resistant. S. flexneri showed the highest resistance (91.2%). S. flexneri serotype 1, resistant to ciprofloxacin and norfloxacin, was observed. All isolates were sensitive to nalidixic acid and ceftriaxone. Only 5 (5.6%) of the isolates were sensitive to all antibiotics tested. Based on these findings, we recommend ciprofloxacin as the drug of choice for treatment of shigellosis in Gondar, with frequent monitoring of drug susceptibility testing. PMID- 19305048 TI - Simulation model of pandemic influenza in the whole of Japan. AB - An influenza pandemic--an outbreak of a novel influenza virus that spreads around the world--is a real threat. To predict the spread of an influenza pandemic across the whole of Japan, this study applies a real individual-based model (ribm) to the whole of Japan and simulates how a single case of influenza could spark pandemic influenza throughout Japan. We used Person-Trip (PT) data for nine regions (the Tokyo metropolitan area, Kansai, Chukyo, Fukuoka, Sendai, Sapporo, Miyazaki, Okinawa, and northern Ibaragi). PT data were collected from randomly chosen persons and contain information on their locations and their uses of all transportation modes including trains, cars, buses, bicycles, and walking. In total, the nine regions have a population of about 72 million, of whom more than 2.20 million individuals participated in the PT surveys. Estimates on the probability of movement among the nine PT regions are based on the results of the Third National Survey for Movement in 2003. Disease transmission in each region or on a train is assumed to occur within a 1-m radius. The approximate numbers of new cases arising on day 14 after the arrival of the first infected person are estimated to be 322,000 in Tokyo, 25,000 in Kansai, 4,800 in Chukyo, 3,600 in Sapporo, 2,600 in Fukuoka, 600 in Sendai, 17 in Okinawa, and 300 in Miyazaki. The disease seems to spread more slowly in smaller cities such as Miyazaki and Okinawa than in big cities such as Tokyo and Kansai. Area quarantines may help to contain outbreaks in smaller cities, but probably not in larger cities. Because this study demonstrates that ribm is useful for simulating the spread of pandemic influenza in Japan, it is hoped that this modeling technique will be used in the creation of preparedness plans. PMID- 19305049 TI - Febrile illnesses of different etiology among outpatients in four health centers in Northwestern Ethiopia. AB - Fever of different etiology is common in tropical and subtropical countries of the world. Etiological agents of febrile illnesses were assessed in 653 acute febrile patients aged 3 to 17 years who attended the outpatient departments of Dembecha Health Center, Jiga Health Center, Quarit Health Center, and Finoteselam Hospital in western Gojjam zone, northwestern Ethiopia. Malaria was the most prevalent illness, infecting 62% of all cases. Its prevalence varied significantly from 52% (Dembecha) to 72.7% (Quarit) (chi(2)=15.02, P=0.000). Plasmodium falciparum was the first cause of malaria (47.3%) followed by P. vivax (23%). Mixed infection of both P. falciparum and P. vivax was found in 7.2% of the cases. The other febrile infections were pneumonia (7%), typhoid (5.8%), typhus (5.1%), and brucellosis (2.6%). The availability of diagnostic facilities and the awareness of the community regarding the prevalence of non-malaria febrile illnesses are very low, and these illnesses are diagnosed clinically. As these illnesses are nonspecific, especially during the early stages of onset, misdiagnosis and mistreatment can occur. Therefore, it is recommended that the necessary diagnostic materials and awareness should be in place for prompt treatment of febrile cases in these districts. PMID- 19305050 TI - HIV/AIDS acquisition and transmission in Bangladesh: turning to the concentrated epidemic. AB - A seventh round behavioral and serological surveillance found that the HIV epidemic had remarkably increased to 7% among intravenous drug users (IDU) in Central Bangladesh, indicating the urgent need to increase prevention. The main purposes of this study were to find out, by collecting data and the necessary information from sero-surveillances, published reports, and articles, what the prevalence of HIV/AIDS is, and what the acquisition and transmission routes are. In addition, trends in HIV-related risk behaviors among recognized high risk groups were observed, and estimations and projections of HIV transmission up to the year 2020 presented. The Estimation and Projection Package was used to estimate and project HIV transmission. The study results reveal that Bangladesh is a low prevalence country which is turning into one with a concentrated epidemic due to the high HIV prevalence rate of IDU (7%) among the most-at-risk groups. Within this at-risk population, IDU have the highest prevalence rate of HIV transmission, followed by female sex workers, clients of sex workers, and men who have sex with men. If the transmission rate continues to increase, the situation will be uncontrolled. Therefore, there is an urgent need for a comprehensive prevention program to control the spread of HIV. PMID- 19305051 TI - The factors associated with asymptomatic carriage of Helicobacter pylori in children and their mothers living in three socio-economic settings. AB - The number of children infected by Helicobacter pylori is increasing worldwide. The aim of this study is to identify demographic and maternal risk factors affecting H. pylori positivity in asymptomatic children. One hundred sixty-five asymptomatic children, 75 (45%) females, and 90 (55%) males, between 2 and 12 years of age (mean 6.8 -/+ 3.0 years) were tested for the presence of H. pylori stool antigen. The ages, genders, weights, heights, and breastfeeding histories of the children were reported. Information concerning the age and education levels of the mothers, number of siblings, and family incomes was also taken. H. pylori stool antigen positivity was 30.9% (n=51) of the children and 30.4% (n=48) of their mothers. H. pylori positivity was detected in 70.6% (36) of children whose mothers were positive (P<0.001, r=0.64). Lower education level of mothers, lower family income, poor living conditions, and higher numbers of siblings were correlated with higher H. pylori positivity in children. The children living in the worst conditions and having less well educated mothers were at higher risk for H. pylori infection. The best way to decrease the prevalence of H. pylori infection in children is to educate women about how to protect themselves and their offspring from H. pylori infection. PMID- 19305052 TI - Spatial analysis and remote sensing for monitoring systems of Oncomelania nosophora following the eradication of schistosomiasis japonica in Yamanashi Prefecture, Japan. AB - In order to develop an inexpensive, simple, and accurate method of monitoring for the reemergence of schistosomiasis japonica in Yamanashi Prefecture, Japan, the distribution and habitation density of the intermediate host, Oncomelania nosophora, were spatially analyzed using geographic information systems. The 1967 1968 density distribution maps prepared by Yamanashi Prefecture and Nihei were digitized and geocoded. The habitats and population density of O. nosophora were estimated by referring to the data compiled by the Yamanashi Association for Schistosomiasis Control (1977). These earlier findings were compared with average population densities between 1996 and 2000 previously recorded (Nihei, N., Kajihara, N., Kirinoki, M., et al., Parasitol. Int., 52, 395-401, 2003 and Nihei, N., Kajihara, N., Kirinoki, M., et al., Parasitol. Int., 53, 199-205, 2004). A variance map was created to compare the spatial distribution maps of population density from each of the two periods of interest. The changes in distribution were remarkable and the map was found to be effective for future control. The most appropriate monitoring sites were chosen on the basis of the spatial population density maps and the variance map. Moreover, the paddy fields at risk were extracted using the normalized difference vegetation index value based on Advanced Land Observation Satellite images. The combination of this method with the global positioning system provides an inexpensive means of monitoring modern schistosomiasis endemic areas in Japan and also in China, the Philippines, and other countries as well, where the intermediate snail grows in paddy fields and marshlands under consistently wet conditions. PMID- 19305053 TI - Lemierre's syndrome followed by acute respiratory distress syndrome successfully rescued by antibiotics and hemoperfusion with polymyxin B-immobilized fiber. AB - Lemierre's syndrome is characterized by a primary oropharyngeal infection in a young healthy person who subsequently develops septic thrombophlebitis of the internal jugular vein and metastatic abscesses. We here report an uncommonly severe case of Lemierre's syndrome with acute respiratory distress syndrome (ARDS), in which polymyxin B-immobilized fiber (PMX) was used as supportive therapy. A 30-year-old, previously healthy man presented with sore throat, fever, rigor, and dyspnea. Chest computed tomography scan revealed multiple bilateral peripheral pulmonary nodules with small bilateral pleural effusions. The patient's condition rapidly deteriorated into ARDS after admission. Intubation followed by mechanical ventilation was required, and hemoperfusion with PMX was useful in alleviating the patient's condition. Isolation of Fusobacterium necrophorum from the blood culture and the contrast-enhanced scan revealed thrombosis and thrombophlebitis in the left internal jugular vein. The patient was diagnosed with Lemierre's syndrome, and an alternative treatment regimen with prolonged administration of ampicillin, clindamycin, and metronidazole resulted in improvement of the patient's respiratory function and general condition. Our case indicated that PMX might be an effective supportive therapy in severe cases of Lemierre's syndrome with ARDS that possessed no indication of surgical interventions. PMID- 19305054 TI - Influenza virus infections in Lebanese children in the 2007-2008 season. AB - We conducted the first epidemiological study of influenza in Lebanon, a temperate country in the Middle East. Between January to May 2008, 39 patients with influenza-like illness were tested. Of these, 51% contracted influenza in January alone, while no influenza cases were detected in May. Among the 39 patients, 11 influenza A and 4 influenza B cases were detected by rapid kit in addition to 10 respiratory syncytial virus cases by real-time PCR. The influenza viruses were genetically divergent from the 2007/2008 season's vaccine strains, but resembled strains circulating in other countries during the same season. PMID- 19305055 TI - First case of treatment failure of artemether-lumefantrine in a Japanese traveler with imported falciparum malaria. AB - Artemether-lumefantrine, a tablet formulation of these respective antimalarial compounds, has been developed for the treatment of patients with drug-resistant malaria worldwide. Many studies have shown that it is most effective of the antimalarial compounds in shortening the fever and parasite clearance times. However, several treatment failures have been reported. These failures are believed to be a consequence of poor bioavailability of the lumefantrine component when ingested without fatty food. This paper reports the first case of such treatment failure of imported malaria in Japan in a 58-year-old Japanese man who showed recrudescence of Plasmodium falciparum after treatment with artemether lumefantrine. The drug was administered to the patient in 6 doses, each time without fatty food and on a seemingly empty stomach. It is believed that treatment failure was due to poor absorption and a subsequent low plasma concentration of lumefantrine. Although artemether-lumefantrine has not been approved for use in Japan and is thus not commonly used there at present, it is thought to be the most promising drug of choice for the treatment of drug resistant malaria. Taking an appropriate dosage and providing patients with proper instructions on taking the drug concurrently with fatty food are required for effective treatment with artemether-lumefantrine. PMID- 19305056 TI - Characterization of p44/msp2 multigene family of Anaplasma phagocytophilum from two different tick species, Ixodes persulcatus and Ixodes ovatus, in Japan. AB - Anaplasma phagocytophilum, which belongs to the order Rickettsiales, is an obligate intracellular bacterium and causes an emerging, tickborne, and febrile infectious disease, anaplasmosis, in humans and other mammals. This bacterium expresses a variety of 44-kDa immunodominant proteins encoded by the p44/msp2 multigene family on the surface for the purpose of avoiding the host immune defense due to the antigenic variation. In Japan, little is known about the molecular and biological features of A. phagocytophilum. In this study, we tried to characterize in detail the p44/msp2 multigene family of A. phagocytophilum from two tick species, Ixodes persulcatus and I. ovatus in Japan. A total of 174 amino acid sequences from the recombinant p44/msp2 clones after TA cloning of the amplicons obtained from the ticks were phylogenetically analyzed. The results showed that most of the clone sequences from I. ovatus were very similar to each other, but the sequences from I. persulcatus were diverse, and the sequences from the ticks were distinct from those from a wild deer that was previously reported. These findings suggest that Ixodes ticks are probably responsible for the transmission of certain genetic variants of A. phagocytophilum and that additional organism selection might occur in I. ovatus. PMID- 19305057 TI - Fungemia and cutaneous zygomycosis due to Mucor circinelloides in an intensive care unit patient: case report and review of literature. AB - Mucor spp. are rarely pathogenic in healthy adults, but can cause fatal infections in patients with immuosuppression and diabetes mellitus. Documented mucor fungemia is a very rare condition in the literature. We described a fungemia and cutaneous mucormycosis case due to Mucor circinelloides in an 83 year-old woman with diabetes mellitus who developed acute left frontoparietal infarctus while hospitalized in a neurological intensive care unit. The diagnosis was made based on the growth of fungi in the blood, skin biopsy cultures, and a histopathologic examination of the skin biopsy. The isolates were identified as M. circinelloides by molecular methods. This case is important in that it shows a case of cutaneous mucormycosis which developed after fungemia and provides a contribution to the literature regarding Mucor fungemia. PMID- 19305058 TI - The evaluation of the Quantiferon-TB Gold test in pulmonary and extrapulmonary tuberculosis. AB - In our study we evaluated the Quantiferon-TB Gold (QFT-G) test and compared it with tuberculin skin test (TST) positivity in 44 pulmonary and 21 extrapulmonary tuberculosis cases in whom active tuberculosis infection was suspected on clinical, microbiologic and radiologic grounds. QFT-G positivity was 75% in pulmonary tuberculosis and 76.2% in extrapulmonary tuberculosis. TST positivity was 68.2 and 62%, respectively. Although QFT-G positivity was higher than TST positivity, the difference was statistically insignificant (P=0.77). Similar to TST, QFT-G is one of the supplementary diagnostic tests for active tuberculosis disease and latent tuberculosis infection. Its advantage over TST is that it gives a result on the same day and it is unaffected by prior BCG and previous exposure to atypical mycobacteria. In conclusion, we think that, as with TST, a positive QFT-G result can be an adjunct to diagnosis in patients having clinical and radiological data compatible with tuberculosis. PMID- 19305059 TI - Carbapenem-resistant Acinetobacter baumannii producing OXA-23 in Thailand. AB - We investigated the resistance determinant of 13 clinical isolates of carbapenem resistant Acinetobacter baumannii collected from a regional hospital in the north of Thailand. All isolates were multidrug resistant and produced the OXA-23 carbapenemase. The bla(OXA-23) gene was found adjacent to ISAba1. Furthermore, two isolates carried the metallo beta-lactamase gene, bla(IMP). The bla(OXA-23) and bla(IMP) genes were plasmid-mediated according to the transformation assays. This is the first description of OXA-23-producing A. baumannii from Thailand. PMID- 19305060 TI - Colonization of a medical center in Southern Taiwan by epidemic strains of carbapenem- and multidrug-resistant Acinetobacter baumannii and the genetic organization of their integrons. AB - A total of 46 carbapenem- and multidrug-resistant (CR- and MDR-)Acinetobacter baumannii bacteremic isolates from a Taiwanese medical center were investigated over the period 2000 to 2006 using randomly amplified polymorphic DNA (RAPD) profiling and by analysing the genetic organization of their integrons. The results of RAPD patterns revealed that before 2003 each CR- and MDR-A. baumannii bacteremic isolate was independent, but after 2003 the isolates appeared to belong in four epidemic strains and persisted in the hospital. All the CR- and MDR-A. baumannii strains harbored class I integron (intI1) genes. PCR amplification and nucleotide sequencing showed that the cassette genes of intI1 were found to form four different antibiotic-resistant gene alignments in those strains. The bla(IMP-1) gene in the cassette genes of intI1 was identified in a clone, which raised great concern that clonal spread of this strain or of an integron-mediated horizontal gene may have occurred. PMID- 19305061 TI - High-level aminoglycoside resistance and beta-lactamase production in enterococci at a tertiary care hospital in India. AB - Enterococci, a family of important opportunistic pathogens, exhibits intrinsic resistance to a number of antimicrobial agents in addition to acquired multidrug resistance. The present study was conducted to determine whether enterococci at a tertiary care hospital in India exhibit high-level aminoglycoside resistance and beta-lactamase production. Enterococci were isolated from various clinical specimens and identified phenotypically. High-level resistance (HLR) to gentamicin, kanamycin, and streptomycin was determined by disc diffusion tests. Beta-lactamase production was detected using three methods: iodometric, acidometric, and chromogenic beta-lactamase assays. Among the 86 enterococci isolated, 34 were found to have HLR to one or more aminoglycosides; HLR to kanamycin was most common. Vancomycin resistance was present in four of the isolates. Only one enterococcus produced beta-lactamase, and it was sensitive to ampicillin on routine disc diffusion testing. Beta-lactamase production among enterococci, though not very common, may be missed on routine susceptibility testing. Frequent occurrence of HLR to kanamycin makes amikacin a poor choice for inclusion in combination therapy with cell wall-active agents. PMID- 19305062 TI - Genotyping of Brucella melitensis by rpoB gene analysis and re-evaluation of conventional serotyping method. AB - In Turkey, where brucellosis is endemic, a comparison of conventional and molecular genotyping methods has not been published to date. In this study, we investigated the efficacy of single nucleotide polymorphism (SNP) analysis of rpoB gene in the genotyping of Brucella melitensis strains by sequencing. In light of the molecular genotyping method available now in Turkey, the adequacy of serological typing alone should be re-evaluated as a tool for epidemiologic studies of B. melitensis. PMID- 19305063 TI - Direct colony PCR of several medically important fungi using Ampdirect plus. AB - Direct colony polymerase chain reaction (DCPCR) is a useful molecular biological technique for application in the field of mycology. In this study, all of the 63 fungal strains examined, including those of the genera Candida and Aspergillus, were amenable to DNA amplification using an Ampdirect(R) Plus kit, which allows direct PCR amplification with no requirement for DNA extraction, following 1 h of rapid fungal lysis. Moreover, we compared DCPCR of 35 strains, representing 20 species, using Ampdirect PCR and standard PCR with no lysis buffer. Thirty-four of these strains (97.14%) yielded positive results on Ampdirect PCR, while only 11 (including Aspergillus fumigatus TIMM1776) of the 35 strains (31.43%) showed PCR products when standard PCR reagents were used. Ampdirect DCPCR was also applicable to DNA amplification for spore and hyphal cells. This approach reduces DNA template preparation time before PCR from fungal colonies, and also reduces the cost of PCR. PMID- 19305064 TI - T serotyping of Streptococcus pyogenes in Aichi Prefecture, Japan isolated between 2003 and 2007. PMID- 19305065 TI - Isolation of Helicobacter cinaedi from a sepsis patient with cellulitis. PMID- 19305066 TI - Toxigenic Corynebacterium ulcerans isolated from the domestic dog for the first time in Japan. PMID- 19305067 TI - Blister aneurysms. PMID- 19305068 TI - Epilepsy surgery in India. AB - This review traces the evolution of epilepsy surgery in India from the beginning to the present state. During the last one and half decades, surgical treatment of epilepsies has made resurgence in this country and at present a few centers have very active and sustained epilepsy surgery programs. Within a 14-year period, the R. Madhavan Nayar Center for Comprehensive Epilepsy Care, Trivandrum, has undertaken over 1000 epilepsy surgeries. However, in the whole country, annually, not more than 200 epilepsy surgeries are currently being performed. This number is a miniscule when compared to the number of potential surgical candidates among the vast population of India. The enormous surgical treatment gap can only be minimized by developing many more epilepsy surgery centers in different parts of our country. PMID- 19305069 TI - Neurology of endemic skeletal fluorosis. AB - Endemic skeletal fluorosis is widely prevalent in India and is a major public health problem. The first ever report of endemic skeletal fluorosis and neurological manifestation was from Prakasam district in Andhra Pradesh in the year 1937. Epidemiological and experimental studies in the endemic areas suggest the role of temperate climate, hard physical labor, nutritional status, presence of abnormal concentrations of trace elements like strontium, uranium, silica in water supplies, high fluoride levels in foods and presence of kidney disease in the development of skeletal fluorosis. Neurological complications of endemic skeletal fluorosis, namely radiculopathy, myelopathy or both are mechanical in nature and till date the evidence for direct neurotoxicity of fluoride is lacking. Prevention of the disease should be the aim, knowing the pathogenesis of fluorosis. Surgery has a limited role in alleviating the neurological disability and should be tailored to the individual based on the imaging findings. PMID- 19305070 TI - Curability of cancer by radiotherapy and chemotherapy, including in neuraxial neoplasms. AB - In the October of 1996, Lance Armstrong, celebrated cyclist and one of the greatest athletes the world has ever seen, at the age of 24, was diagnosed with metastatic testicular cancer with disease having already spread to his abdomen, lungs and brain. Lance underwent four cycles of chemotherapy, actually the pretty standard one, pioneered at the Indiana University and not only did he get completely cured of his cancer, he remains extremely well till date, 12 years later. He sure did have a few adverse effects during those cycles of chemotherapy in the form of nausea, vomiting, weakness and fall in blood count but he knew and experienced them only for a short transient time and emerged triumphant and strong. In fact, he went on to win six awe-inspiring and incredible successive Tours de France victories from 1999-2005, one of the most grueling sporting events testing the endurance of the very fittest. After his retirement, he has been so inspired that he has completely devoted himself to educate people about the common myths about cancer, and promised to raise awareness and generate money for furthering research into surgery, radiation therapy and chemotherapy for cancer through his foundation. He says "I am indebted to the doctors, nurses and medicine and would want to pay them back for all their energy and caring." In his successful journey of overcoming cancer, he captures the essence of its treatment so well by declaring "Pain is temporary, it may last a minute, or an hour, or a day, or a year, but eventually, it will subside and something else will take its place. If I quit, however, it will last forever". PMID- 19305071 TI - Re-canalization in acute ischemic stroke: the strategies. AB - Thrombolytic stroke therapy is based on the "re-canalization hypothesis," i.e., that reopening of occluded vessels improves clinical outcome in acute ischemic stroke through regional reperfusion and salvage of threatened tissues. Re canalization is an important predictor of stroke outcome in all the modalities of thrombolysis. Thrombolysis in acute ischemic stroke evolved from clinical trials with intravenous (IV) tissue plasminogen activator (tPA) to combination treatments with Intra-arterial (IA)/mechanical reperfusion techniques. The combined approach reduces time to initiation of treatment and may increase re canalization and chances of a good clinical outcome. In IV thrombolysis overall re-canalization rate is 46.2% during the first 6-24 hrs. Intra-arterial thrombolysis has higher early re-canalization rate, 63.2%. The highest re canalization rate is seen with mechanical thrombolysis, 83.6%. Other factors in addition to re-canalization: stroke severity, older age, systolic hypertension, location of arterial occlusion, collateral blood supply, and time from stroke onset to treatment play an important role in determining the clinical outcome following thrombolysis with tPA. Future trials should look at the interplay of various factors like re-canalization, reperfusion, and collateral flow and clot burden in the evaluation of reperfusion therapies in acute ischemic stroke. PMID- 19305072 TI - Expressions of Fas/DcR3 and RGD-FasL mediated apoptosis in pituitary adenomas. AB - PURPOSE: To detect the expressions of Fas/DcR3 and to investigate the cytotoxic effects of RGD-FasL on pituitary adenoma cells. MATERIALS AND METHODS: Fas/DcR3 mRNAs were detected by Reverse transcription polymerase chain reaction (RT-PCR) and their surface expressions were measured by flow cytometry. Cytotoxicities exerted by FasL and newly-constructed RGD-FasL on tumor cells were measured with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The apoptotic cells were examined by electron microscopy and the induced apoptosis was determined by agarose gel electrophoresis. The cell cycle was assessed by flow cytometry with ANNEXIN V FITC/PI. The expressions of caspases, Bcl-2, RANKL and JNK2 were detected by Western blotting. RESULTS: Fas/DcR3 was expressed in GH3/MMQ/AtT20 cells. The cytotoxic effects of RGD-FasL on tumor cells were seen in a dose-dependent manner. These cells showed the same sensitivity to RGD-FasL as to FasL. RGD-FasL induced apoptosis and G1/G0 arrest. The expressions of caspase-8/9/3, RANKL, JNK2 were increased while that of Bcl-2 was decreased with treatment of RGD-FasL. CONCLUSIONS: Fas can be a novel target for the treatment of pituitary adenomas. RGD-FasL induces apoptosis of pituitary adenoma cells through caspase activation. PMID- 19305073 TI - C1 and C2 nerve sheath tumors: analysis of 32 cases. AB - BACKGROUND: C1 and C2 nerve sheath tumors are to be considered as a separate clinical entity because of their unique clinical presentation, relation to the adjacent structures and surgical approaches when compared to their counterparts in other regions. AIM: To present the clinical characteristics, radiological findings, operative approaches, and outcomes. SETTING AND DESIGN: Of the 32 patients, the study was retrospective in 22 patients and in 10 it was prospective. MATERIALS AND METHODS: Patients' case records were analyzed. Diagnosis was established by magnetic resonance imaging of craniovertebral junction. Patients were evaluated by Yasargil grading and operative procedure was done accordingly. RESULTS: In this series the tumors were multiple and had extradural extension and hourglass expansion. Surgical results were excellent in most patients when procedure was done early in the course of the disease. Best results in terms of complete removal and good neurological outcome were achieved with posterior or posterolaterlly located lesion. Of the 20 patients who had surgery by posterior approach, 19 (95%) patients had improvement postoperatively, while one (5%) patient remained unchanged. While of the 8 patients who had surgery by posterolateral approach, 6 (75%) had improvement postoperatively, one had no change and the other had neurological deterioration. CONCLUSION: C 1 and C 2 nerve sheath tumors are to be considered as a separate clinical entity. Operative results and recovery of patients are excellent when tumor is located posterior or posterolaterally and surgical procedure is done early in the disease course. PMID- 19305074 TI - Epstein-Barr virus as a possible etiologic agent in primary central nervous system lymphoma in immunocompetent individuals. AB - BACKGROUND: Primary central nervous system lymphoma (PCNSL) occurs in both immunocompetent and immunosuppressed individuals. The role of Epstein-Barr virus (EBV) has been implicated in immunosuppressed individuals but its role is not established in immunocompetent individuals. AIMS: To study the possible role of EBV in PCNSL in immunocompetent individuals. SETTING AND DESIGN: Retrospective study. MATERIALS AND METHODS: Thirty patients with PCNSL were studied immunohistochemically with antibodies to CD45, CD20, CD3 and EBV latent membrane protein-1 (EBV LMP-1). In situ hybridization was done in 19 patients where enough tissue was available using a specific oligonucleotide probe for EBV-Early RNA (EBER). RESULTS: All the patients were immunocompetent and mean age was 41.6 years. Histologically they were diffuse large cell lymphoma: 25 (83.3%) were B cell, 1(3.3%) was T cell and 4 (13.3%) were unclassified. EBV LMP-1 showed variable membrane and cytoplasmic positivity in 24 (80%) patients. In situ hybridization for EBER was negative in all the 19 patients studied. CONCLUSION: In this region of the world probably EBV has no etiologic role in PCNSL in immunocompetent individuals. PMID- 19305075 TI - Ullrich congenital muscular dystrophy: report of nine cases from India. AB - BACKGROUND: Ullrich congenital muscular dystrophy (UCMD) is a unique congenital disorder characterized clinically by generalized muscle weakness, contractures of the proximal joints and hyperextensibility of the distal joints and begins from birth or early infancy. MATERIALS AND METHODS: We prospectively evaluated nine cases of classical UCMD and recorded the clinical phenotypic characteristics and the histopathological findings. RESULTS: There were eight boys and one girl child with classical features of severe muscle weakness, prominent proximal contractures, distal hyperlaxity and prominent calcanei. Immunohistochemistry for Collagen VI A1 done on seven cases showed total absence of labeling in six while sarcolemmal-specific deficiency was noted in one case confirming the diagnosis of UCMD. Interestingly, all our patients were noted to have near total absence of major palmar and plantar creases, and instead there were fine mesh-like lines in addition to the soft velvety skin on the palms and soles suggestive of altered collagen arrangements in the skin. Hitherto, this clinical finding has not been described in UCMD in the English literature. PMID- 19305076 TI - Variation in the origin of the posterior cerebral artery in adult Sri Lankans. AB - BACKGROUND: The degree of contribution from the vertebro-basilar and carotid systems to the origin of the posterior cerebral artery (PCA) is of clinico anatomical importance. Three configurations in the bifurcation of the posterior communicating artery (PcomA) have been described, foetal, transitional and adult. OBJECTIVE: To examine the extent of contribution from the vertebro-basilar and carotid systems to the origin of the PCA in adult autopsy brains in subjects who had died of causes unrelated to the brain. MATERIALS AND METHODS: The external diameter of the PcomA, pre communicating part (P1) and the post communicating part (P2) of PCA in 225 normal Sri Lankan adult cadaver brains was measured using a calibrated grid. RESULTS: The adult configuration was present in 220 (93.3%); (bilaterally in 200 (88.8%), and unilaterally in 20 (8.8%) ten on each side. Foetal configuration was seen in 17 (4.4%); bilaterally in 3 (1.3%), and unilaterally in 14 (6%) 8 on the left and 6 on the right. Transitional configuration was seen in 8 (2.2%); bilaterally in 2 (0.9%), and unilaterally in 6 (2.5%) 2 on the left and 4 on the right. CONCLUSION: This study reveals that the internal carotid artery (ICA) provided the major supply to the PCA in 4.4% of foetal and 2.2% of transitional configurations. The adult configuration was present in 220 (93.3%), the highest recorded in the literature. This finding may be of relevance to vertebrobasilar ischaemia and infarcts in the territory of the PCA. PMID- 19305077 TI - D-dimer is useful in the diagnosis of cortical venous sinus thrombosis. AB - BACKGROUND: Estimations of D-dimer correlate with deep vein thrombosis and pulmonary thromboembolism and may serve as a marker of cortical venous sinus thrombosis (CVST). AIM: To study the usefulness of D-dimer in the diagnosis of CVST. MATERIALS AND METHODS: A total of 26 patients with CVST were subjected to a detailed medical history and neurological evaluation. A cranial MRI was carried out on a 1.5T scanner using T1, T2, and DWI sequences and two-dimensional time of flight MR venography. D-dimer was estimated using a rapid latex agglutination slide test using monoclonal antibodies. RESULTS: The age of patients ranged between 16 and 70 years old (median 31 years old); 8 were males and were examined after a mean duration of 22 days of symptoms. Cortical venous thrombosis was attributed to puerperium in 4 patients, infection in 5 patients, and pregnancy, dehydration, insect bite, and migraine in 1 patient each. Superior sagittal sinus was involved in 15 patients, transverse sinus in 16 patients, straight sinus in 3 patients, sigmoid sinus in 7 patients, and cavernous and deep system in 1 patient each. A total of 12 patients had more than one sinus involvement. D-dimer was positive in 20 patients and correlated with the duration of symptoms but not with the extent of sinus thrombosis and the outcome. CONCLUSION: D-dimer is useful in patients suspected of CVST and patients with positive test results should be urgently sent for MR imaging. PMID- 19305078 TI - Remission of concomitant Henoch-Schoenlein purpura and Sydenham chorea after intravenous corticosteroids. AB - We report a young girl who developed Henoch-Schoenlein purpura at the age of 11 years. Two weeks later she developed severe asymmetric choreic movements and behavioral disturbances. Sydenham s chorea was diagnosed based on the laboratory evidence and she was given intravenous methylprednisolone for five consecutive days. Both behavioral and movement disorder rapidly resolved. She was asymptomatic at three years of follow-up. The rapid resolution of choreic movements and behavioral disturbances in our patients suggests, intravenous corticosteroids may be an option in the treatment of Sydenham's chorea, more so when the movements are disabling. PMID- 19305079 TI - Prosopagnosia: a rare presenting manifestation of frontotemporal lobar degeneration. AB - Frontotemporal dementia is an important neurodegenerative disorder accounting for a significant proportion of dementia cases with onset before 60 years of age. Apart from the well recognized behavioral changes the disease has many other distinctive features like predominant language involvement alone or associated features of motor neuron disease or parkinsonism etc. which at times may be the presenting manifestation itself. In the following article we describe a rare presenting manifestation; prosopagnosia, in the setting of frontotemporal degeneration. PMID- 19305080 TI - Uncommon presentation of giant cell arteritis: report of two cases with scalp necrosis. AB - Giant cell arteritis (GCA), the most common form of systemic granulomatous vasculitis in adults, preferentially involves large and medium-sized arteries. Scalp necrosis is a rare complication and can be the presenting feature. This report presents two patients of GCA with severe scalp necrosis as the presenting symptoms. Both the patients were treated successfully with selective debridement, local wound care, antimicrobial and systemic steroid treatment. Skin necrosis is a potentially treatable complication of GCA and needs a multidisciplinary approach. PMID- 19305081 TI - Reversible cerebral vasoconstriction syndrome. AB - Reversible cerebral vasoconstriction syndromes (RCVS) are a group of disorders that have in common an acute presentation with headache, reversible vasoconstriction of cerebral arteries, with or without neurological signs and symptoms. In contrast to primary central nervous system vasculitis, they have a relatively benign course. We describe here a patient who was diagnosed with RCVS. PMID- 19305082 TI - Balo's concentric sclerosis presenting with benign clinical course and multiple sclerosis-like lesions on magnetic resonance images. AB - Balo's concentric sclerosis (BCS) is a rare primary demyelinating disease of central nervous system (CNS) and is considered to be a variant of multiple sclerosis (MS). It is characterized by a severe, rapidly evolving course with CNS lesions consisting of concentric rings of demyelination alternating with myelination in the white matter. We report a patient with BCS from mainland China diagnosed on magnetic resonance imaging (MRI) findings. In addition to two contrast enhanced open ring lesions. MRI also showed multiple small white matter lesions in both the hemispheres. The patient had a good response to corticosteroids and had a benign prognosis during a follow-up period of two years. PMID- 19305083 TI - The influence of hypoglycemia on cerebral blood flow in cases of hypoglycemic coma. AB - The authors report two cases of hypoglycemic coma in which both diffusion weighted imaging (DWI) and single photon emission computed tomography (SPECT) were used. A 74-year-old woman (case 1) presented with deep coma associated with hypoglycemia (blood glucose level: 20 mg/dl), and DWI on admission revealed extensive hyperintensity lesions. SPECT obtained on the second hospital day revealed diffuse hyperperfusion and a follow-up SPECT on the eighth hospital day showed relative hypoperfusion. An 89- year- old woman (case 2) presented with coma associated with hypoglycemia (blood glucose level: 45 mg/dl), and DWI showed diffuse hyperintensity lesions in both hemispheres. SPECT obtained on the ninth hospital day showed relative hypoperfusion compared to the first scan obtained on the third hospital day. Although the blood glucose level was rapidly corrected, the level of consciousness remained unchanged in both cases. Our results suggest that cerebral blood flow may increase even after glucose correction and that relative hypoperfusion could occur in the subacute stage of hypoglycemic coma. PMID- 19305084 TI - Brain abscess in a non-penetrating traumatic intracerebral hematoma: case report and review of literature. AB - We report a 57-year-old man who presented one month after sustaining a traumatic right temporal intracerebral hematoma with history of headache, left hemiparesis and altered sensorium of two days duration. A diagnosis of right temporal resolving hematoma was made on computed tomography scan. However, his sensorium progressively deteriorated and he underwent craniotomy and partial excision of an abscess. He was treated with appropriate antibiotics for six weeks despite of which he did not improve and died nine months later. We conclude that there should be a high index of suspicion for brain abscess in patients with traumatic intracerebral hemorrhage if the clinical and radiological picture is different from the expected course of a resolving hematoma. PMID- 19305085 TI - Olfactory ensheathing cell tumor. AB - Olfactory ensheathing cells (OECs) are found in the olfactory bulb and olfactory nasal mucosa. They resemble Schwann cells on light and electron microscopy, however, immunohistochemical staining can distinguish between the two. There are less than 30 cases of olfactory groove schwannomas reported in the literature while there is only one reported case of OEC tumor. We report an OEC tumor in a 42-year-old male and discuss the pathology and origin of this rare tumor. PMID- 19305086 TI - Persistent primitive hypoglossal artery associated with lower basilar artery aneurysm: diagnosis and clinical implications. AB - An elderly male was evaluated for subarachnoid hemorrhage and found to have a persistent primitive hypoglossal artery with its associated abnormal posterior fossa circulation and a lower basilar artery aneurysm. Angiography findings are described and are corroborated with computed tomography and magnetic resonance imaging. Its surgical and clinical relevance are discussed. PMID- 19305087 TI - Metastatic adenocarcinoma of bilateral cavernous sinus and optic nerve with unknown primary mimicking orbital pseudotumor. AB - Bilateral cavernous sinus and optic nerve metastases is extremely rare. We report an extremely rare case of metastatic adenocarcinoma of bilateral cavernous sinus and optic nerve with unknown primary presenting as orbital pseudotumor. She underwent bilateral optic nerve decompression and the diagnosis was established by biopsy. PMID- 19305088 TI - Ruptured and unruptured aneurysms of the accessory anterior cerebral artery combined with a blood blister-like aneurysm of the anterior communicating artery. AB - Anterior communicating artery (Acom) complex and anterior cerebral artery (ACA) are the frequent sites of arterial anomalies. Aneurysms associated with triplicate ACA are rare. Blood blister-like aneurysm (BBA) of the Acom is also very rare. This report presents these two rare types of aneurysms in a 63-year old man who presented with subarchnoid hemorrhage secondary to accessory ACA aneurysmal rupture. During surgery, a BBA of the Acom was also found. Both the aneurysms were treated simultaneously. The BBA bled intraprocedurally but was repaired successfully by neck reconstruction and clipping with the preservation of the parent and the branching arteries. The ruptured accessory ACA aneurysm was clipped together with the artery which had multiple aneurysms distally. For the treatment of a complex aneurysm like this, accurate understanding of the anatomic variations, proper imaging study and extremely careful technique are essential. PMID- 19305089 TI - Spinal epidural cavernous hemangioma with myelopathy: a rare lesion. AB - A case of pure epidural cavernous angioma involving the dorsal region of the spine is presented. The clinical picture was of a slowly progressive compressive myelopathy. Imaging studies only showed a dorsally located epidural mass lesion. Histology confirmed the diagnosis of this rare type of lesion. The patient improved after excision of the lesion. PMID- 19305090 TI - Posterior circulation stroke in a cannabis abuser. PMID- 19305091 TI - Cerebral venous thrombosis due to homozygous factor V Leiden mutation. PMID- 19305092 TI - Isolated brain metastasis from malignant melanoma of choroid seven years following enucleation. PMID- 19305093 TI - Tardive dyskinesia following risperidone treatment in Tourette's syndrome. PMID- 19305094 TI - Primary stenting in acute carotid dissection. PMID- 19305095 TI - Multiple extracranial metastases from intradiploic meningioma. PMID- 19305096 TI - Ruptured intracranial dermoid cyst. PMID- 19305097 TI - Extradural thoracic spinal meningioma. PMID- 19305098 TI - Giant hemispheric multiloculated brain abscess. PMID- 19305099 TI - Cavernous sinus thrombophlebitis causing reversible narrowing of internal carotid artery. PMID- 19305100 TI - The hot cross bun sign. PMID- 19305101 TI - Simultaneous single cell stable expression of 2-4 cDNAs in HeLaS3 using psiC31 integrase system. AB - An important consideration in the design of multigene delivery technology is the availability of suitable vectors to introduce multiple genes stably and stoichiometrically into living cells and co-express these genes efficiently. As a promising system for this purpose, we developed multi-cDNA expression constructs harboring two to three tandemly situated cDNAs in a single plasmid. The utility of this vector system is amplified by combining it with the psiC31 recombinase system which mediates site-specific integration of the genes into naturally occurring chromosomal sequences. By analyzing 55 psiC31-mediated integration events with five different constructs, each carrying one, two or three tandem cDNA expression cassettes, we identified 39 pseudo attP sites in the HeLaS3 chromosomes. All these sites share a common motif containing an inverted repeat and showing a similarity to the native psiC31 attP. The 36 integration events represented 27 different pseudo attP sites, suggesting the possibility of duplicate integration of the multigene expression plasmids into different genomic loci in a single cell. We demonstrated successive introduction of two different multi-cDNA expression plasmids into definite chromosomal pseudo attP sites, attaining integration of four cDNAs of known genomic constitution at precise genomic loci of a single HeLaS3 cell. The expression levels of these several transgenes were enhanced and made equally stable and robust by inserting the cHS4 insulator between genes. PMID- 19305102 TI - Noninvasive diagnosis of culprit lesions with the use of single-photon emission computed tomography/computed tomography fusion images in patients with multivessel coronary artery disease. PMID- 19305103 TI - Drug-induced interstitial lung diseases associated with molecular-targeted anticancer agents. AB - Little was known about drug-induced interstitial lung disease (ILD) when acute ILD-type events developed in several Japanese patients treated with gefitinib. A better understanding of drug-induced ILD is required, including more reliable data about the incidence of events associated with different treatments and identification of the risk factors for this type of ILD. Recent advances in imaging, molecular examination, and pathology have been used in postmarketing surveillance studies designed and conducted by an independent academic team to define the risk and to increase the amount of evidence about ILD related to various molecularly targeted anticancer agents. These studies may shed light on the underlying mechanisms of drug-induced ILD and appropriate evidence-based strategies that can be used to prevent or manage these events. PMID- 19305104 TI - Effect of lidocaine tape on pain during intralesional injection of triamcinolone acetonide for the treatment of keloid. AB - BACKGROUND: Because intralesional injection of triamcinolone acetonide (TA), a widely used for the treatment of keloid, is painful, many patients discontinue treatment. We evaluated the effects of pretreatment with topical 60% lidocaine tape on the pain and tolerability of intralesional TA treatment in patients with keloid. METHODS: The subjects were 42 patients with keloid who had been treated with intralesional injection of TA but had discontinued treatment owing to intolerable pain. All patients were pretreated with 60% lidocaine tape placed on the keloids for more than 120 minutes before intralesional injection of TA. Patients assessed pain with a 100-mm visual analog scale (VAS) with 0 mm for "no pain" and 100 mm for "worst possible pain." Pain was assessed with the VAS immediately after TA injection. Finally, the patients assessed the tolerability of this treatment. RESULTS: The mean VAS score during intralesional TA injection therapy without pretreatment with lidocaine tape was 82.6 +/- 14.4 mm. In contrast, the mean VAS score during intralesional TA injection therapy in the same patients after pretreatment with lidocaine tape was 18.9 +/- 11.3 mm, which was significantly lower (P<.0.05), and 30 (71.4%) of the patients tolerated this therapy well. CONCLUSION: Pretreatment with 60% lidocaine tape significantly reduces the pain associated with intralesional injection of TA. This approach increases patient comfort and should enable patients to continue the treatment. PMID- 19305105 TI - A case of an intramesosigmoid hernia. AB - We report an extremely rare case of an intramesosigmoid hernia with small bowel herniation in a defect on the right (medial) leaf of the mesosigmoid. A 46-year old man was admitted to the hospital complaining of lower abdominal pain, nausea, and vomiting for 6 days. He had undergone an operation for a right inguinal hernia and an appendectomy during childhood. An abdominal X-ray film obtained at admission showed small bowel gas with niveau formation which was diagnosed as small-bowel obstruction. A decompression tube was immediately inserted, and the symptoms subsided. Enterography revealed two strictures separated by approximately 10 cm. However, the contrast medium flowed smoothly through the anal side of the strictures. After the decompression tube was removed, small bowel obstruction recurred, and laparotomy was performed on the 18th day after admission. During the operation, small bowel herniation with a 4 x 3-cm defect was found on the right leaf of the mesosigmoid, and intramesosigmoid hernia was finally determined to be the cause of the small-bowel obstruction. The resection of the incarcerated part was necessary because a large amount of scar tissue was present on the surface. The postoperative course was uneventful, and no recurrence was observed after discharge. A review of this case indicated that the diagnosis might have been successfully obtained with enterography. Although we did not choose laparoscopic surgery, this surgical modality may also be an appropriate treatment for this disease. PMID- 19305106 TI - Frontal musculocutaneous V-Y island flap for coverage of forehead defect with a dural exposure after craniotomy. AB - We have performed forehead reconstruction with a frontal musculocutaneous V-Y island flap to establish skin and soft-tissue coverage for a cranial bone defect with dural exposure. A 56-year-old woman who had previously undergone craniotomy for aneurysm clipping had a severe infection of the bone flap and subsequently underwent partial resection. The skin defect and the underlying dead space on the dura was successfully covered with a frontal musculocutaneous V-Y island flap without complications. Because this flap shows technical feasibility in harvesting, stable blood supply, functional preservation of frontal muscle, and good texture and color match, it may be an ideal flap for forehead and frontal reconstruction of defects of small or moderate size when primary closure, skin grafting, or transfer of local pedicled flaps or free flaps is impossible. PMID- 19305107 TI - Identification of 6 sentinel lymph nodes in a case of malignant melanoma on the ungual region of the finger. AB - We describe a case of malignant melanoma on the ungual region of the index finger with which 6 sentinel lymph nodes were detected in the axillary basin. Of these nodes, 1 of 3 brachial nodes and 2 of 3 central axillary nodes showed micrometastasis. Additional lymph nodes obtained at the subsequent complete axillary lymph node dissection showed no metastasis. Identification of as many as 6 sentinel lymph nodes was ascribed to a single lymph trunk that would branch off to afferent lymphatics draining to each nodal group; however, a gamma probe may have detected radioactive tracer passing to the sequential nodes as well as that remaining in true sentinel nodes. PMID- 19305108 TI - A case of Takotsubo cardiomyopathy during 5-fluorouracil treatment for rectal adenocarcinoma. AB - A case of acute heart failure due to Takotsubo cardiomyopathy induced by 5 fluorouracil is described. Acute heart failure developed during the administration of 5-fluorouracil (5-FU) and levofolinate calcium in a 62-year-old woman who had underwent a Miles operation for rectal adenocarcinoma. Electrocardiography upon admission showed slight ST elevation in leads V1-3, and follow-up electrocardiography on the third hospital day evealed QT interval prolongation and giant negative T waves in leads II, III, aVF, and V1-6. Echocardiography and myocardial scintigraphy showed left ventricular apical ballooning in the acute phase of heart failure, but left ventricular contraction was normal during the recovery phase. Coronary angiography demonstrated normal coronary arteries, and multi-vessel coronary artery vasospasms including microcirculation disorders could be provoked by intracoronary acetylcholine infusion during, but not before, the intravenous administration of levofolinate calcium and 5-FU. The cause of heart failure in this patient, Takotsubo cardiomyopathy induced by multivessel coronary vasospasm including microcirculation disorders only during 5-FU administration, is notable. PMID- 19305109 TI - Incisional bladder hernia with temporary bowel incarceration: report of a case. AB - Abdominal hernias are not rare in women, but incisional bladder hernias are rare. The incisional hernia is a condition caused by protrusion of the abdominal viscera through the abdominal fascia. The presenting symptoms in the cases reported included suprapubic discomfort, irritative voiding symptoms, and urinary incontinence. We present a case of bladder herniation with temporary bowel incarceration through a lower midline incision, which followed operative intervention. The temporary bowel herniation was managed conservatively because the impairment of the blood supply was not severe. PMID- 19305110 TI - Heterogeneous pulmonary vein myocardial cell repolarization and heterogeneous sympathetic hyperinnervation as the substrates for atrial fibrillation. PMID- 19305111 TI - Care of the victims of the Akihabara massacre. PMID- 19305112 TI - Study of MicroRNA expression profiles of esophageal cancer. PMID- 19305113 TI - The anticancer effect of histone deacetylase inhibitors and combination with the cytotoxic agents in lung cancer cells: biological analyses for future clinical application. PMID- 19305114 TI - Prevention of postoperative infection and micrometastasis by preoperative administration of a PPARgamma agonist following colorectal cancer surgery. PMID- 19305115 TI - Gender-related differences in circulating levels of adipocytokine associated with postoperative complication following major surgery. PMID- 19305116 TI - Physician job satisfaction and working conditions in Japan. AB - OBJECTIVES: The aim of this study was to determine factors of working conditions associated with job satisfaction among physicians in Japan. METHODS: We sent a questionnaire to all the physicians who graduated from a medical school in Japan. Physicians who were satisfied with their job were determined as those who selected "very satisfied" and "satisfied" in response to the question: "Overall, are you satisfied with your job?" Working conditions were determined from 10 different aspects: income fairness, hospital resources, career satisfaction, difficulty in patient care, lack of personal time, administrative work, workload, and relationships with physician colleagues, staff and patients. Logistic regression analysis was used to explore the association between working conditions and job satisfaction. RESULTS: Among the respondents, 209 (55.4%) men and 62 (61.4%) women were determined to be satisfied with their job. Job satisfaction was associated with income fairness for both men (corrected odds ratio 1.31, 95% confidence interval 1.09 to 1.47) and women (1.35, 1.05 to 1.53). For men, job satisfaction was associated with good hospital resources (1.45, 1.29 to 1.57), high career satisfaction (1.41, 1.23 to 1.57), good relationships with physician colleagues (1.33, 1.12 to 1.49), and good relationships with hospital staff (1.28, 1.07 to 1.45). For women, job satisfaction was associated with good relationships with patients (1.41, 1.07 to 1.56). CONCLUSIONS: Certain working conditions were important factors for job satisfaction among physicians. These factors should be discussed for improving working conditions. PMID- 19305117 TI - Limitations on work and attendance rates after employees with cancer returned to work at a single manufacturing company in Japan. AB - OBJECTIVES: The purpose of this study was to evaluate the work limitations and attendance rates after employees diagnosed with cancer returned to work from sick leave, and to identify the related factors for the limitations and attendance rates at a single manufacturing company in Japan. METHODS: This study retrospectively analyzed 129 men and 4 women, employed in a single manufacturing industry, who returned to work after sick leave due to newly diagnosis of cancer. Limitations on work after the return to work were enforced in the workplace based on an industrial physician's evaluation. All the employees who needed measures for work were examined by the industrial physicians every 1-6 months until the termination of such work limitations. RESULTS: Limitations on work after the return to work were enforced for 79 (59%) employees (36 employees with alteration of work, 31 with prohibition of shift work and 55 with prohibition of overtime work). A higher degree of work limitations was significantly correlated with work related factors before sick leave (i.e. shift work, production line) as well as disease/treatment-related factors (i.e. chemotherapy, recurrence/metastasis), while the attendance rates after the return to work were not correlated with adverse work-related factors before sick leave. CONCLUSION: The enforcement of work limitations for employees with cancer was relatively common and was based on both disease/treatment- and work-related factors, and this phenomenon may play an important role in the return to work as well as the successful continuation of work after cancer survivors return to work. PMID- 19305118 TI - Assessment of genotoxic damage in nurses occupationally exposed to anaesthetic gases or antineoplastic drugs by the comet assay. PMID- 19305119 TI - Employment contracts: cross-sectional and longitudinal relations with quality of working life, health and well-being. AB - OBJECTIVES: The aim of this study was to enhance (i) insight in the relationship between different types of employment contract and the quality of working life, health and well-being, and (ii) our causal understanding of these relationships by comparing employees whose contract type changes across time. METHODS: Analyses were based on a two-year prospective cohort study. Cross-sectional analyses were based upon a sample of 2,454 Dutch employees (2004). Longitudinal data were available for 1,865 respondents (2004-2006). We distinguished among 5 contract types, and subgroups of 'Upward' (i.e., towards permanent employment) and 'Downward' (towards temporary employment) movers across time. Data were analysed with analysis of variance and cross table analysis. RESULTS: Cross-sectionally, we found differences between contract types in quality of working life: generally permanent employees had better jobs, whereas temporary agency workers and on call workers had more 'bad work characteristics'. We also found a difference in health behaviour (smoking) and that psychological health was worst among temporary agency workers. In longitudinal analyses we found some evidence that a positive change in employment contract was associated with a better quality of working life and better psychological health, whereas the opposite was true for a negative contract change. CONCLUSIONS: The quality of working life, health and well-being are unequally distributed over employment contract groups. Temporary agency workers and on-call workers deserve special attention in terms of job design and human resource management. PMID- 19305120 TI - Protective effects of pioglitazone against global cerebral ischemic-reperfusion injury in gerbils. AB - Despite of the huge socio-economic burden, stroke still represents an unmet therapeutic need. Researchers failed to reproduce preclinical efficacy in subsequent clinical development. To bridge this translation failure, the Stroke Therapy Academic Industry Round Table (STAIR) has suggested a rigorous, robust, and detailed preclinical evaluation in at least 2 species and multiple cerebral ischemia models to avoid the clinical failure. Considering these recommendations, in the present study, we have investigated the effects of pioglitazone in global model cerebral ischemic-reperfusion (IR) injury in gerbils. Global cerebral IR injury, produced by bilateral carotid artery occlusion for 5 min, was characterized by neurological deficits, hyperlocomotion, and neurodegeneration in the hippocampal CA1 region. Global ischemia was also associated with oxidative stress and DNA fragmentation as evident from increased malondialdehyde (MDA) levels and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling)-positive cells. Global cerebral IR injury associated neurological damage was significantly attenuated by pioglitazone pretreatment as evident from reduction in neurological symptoms, hyperlocomotion, and CA1 hippocampal neuronal damage in IR-challenged gerbils. Pioglitazone pretreatment also attenuated the oxidative stress and DNA fragmentation after cerebral IR injury. Pioglitazone post-treatment has also significantly reduced the CA1 hippocampal neuronal damage and DNA fragmentation after cerebral IR injury in IR-challenged gerbils. This study demonstrates the neuroprotective activity of pioglitazone in global cerebral IR injury and its neuroprotective effects may be attributed to reduction in oxidative stress and DNA fragmentation. PMID- 19305121 TI - Panax notoginseng Attenuates the Infarct Volume in Rat Ischemic Brain and the Inflammatory Response of Microglia. AB - The roots of Panax notoginseng (PN) are commonly used as a therapeutic agent to stop hemorrhage and as a tonic to promote health in traditional Korean medicine. Stroke triggers an inflammatory response that not only plays a central role in the pathogenesis of cerebral ischemia, but also induces secondary damage. This study was designed to investigate the neuroprotective effects of the methanol extract of PN on the infarct volume induced by middle cerebral artery occlusion (MCAO) (90-min occlusion and 24-h reperfusion) in rat brains. The PN extract (50 mg/kg, i.p.) was administered 2 h after the onset of MCAO. The PN-treated groups had a reduction in infarct volume by 23.82 +/- 8.9%. In the PN extract-treated groups, the microglial density was significantly decreased in the peri-infarct region; the underlying mechanism was inhibition of inflammatory mediators, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, via blocking of the NF-kappaB pathway. Furthermore, in vitro studies showed that the PN extract significantly reduced the production of iNOS-derived NO and COX-2 derived prostaglandin E(2) through the regulation of gene transcription levels in primary microglia and BV-2 cells. These results suggest that anti-inflammatory and microglial activation inhibitory effects of the PN extract may contribute to its neuroprotective effects in brain ischemia. PMID- 19305122 TI - Cordycepin attenuates neointimal formation by inhibiting reactive oxygen species mediated responses in vascular smooth muscle cells in rats. AB - We determined the action mechanism of cordycepin, a major bioactive component of Cordyceps militaris, on responses of rat aortic smooth muscle cells (RASMCs) and on vascular disorders, especially neointimal formation. Cordycepin inhibited platelet-derived growth factor-BB (PDGF-BB)-induced RASMCs migration and proliferation in a dose-dependent manner. However, pre-treatment with N(omega) nitro-L-arginine methyl ester, a nitric oxide synthase (NOS) inhibitor, and 1,3 dipropyl-8-sulphophenylxanthine (DPSPX), an A(1)/A(2) adenosine-receptor antagonist, abolished the inhibitory role of cordycepin. Cordycepin suppressed the phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) and heat shock protein 27 (Hsp27), but not that of extracellular signal-regulated kinase (ERK) 1/2 in RASMCs stimulated by PDGF-BB. The production of reactive oxygen species (ROS), O(2)(-) and H(2)O(2), induced by PDGF-BB was abolished by the treatment of cordycepin. Moreover, the sprout outgrowth of aortic rings by PDGF BB was inhibited by cordycepin. In vivo neointimal formation evoked by balloon injury was significantly attenuated by the administration of cordycepin. These results demonstrate that cordycepin may exert inhibitory effects on PDGF-BB induced migration and proliferation via interfering with adenosine receptor mediated NOS pathways, thus resulting in the attenuation of neointima formation. In conclusion, cordycepin may be a potent, promising anti-atherosclerosis agent. PMID- 19305123 TI - Characteristic effects of anti-dementia drugs on rat sleep patterns. AB - The present study was undertaken to clarify the effects of anti-dementia drugs on sleep pattern in rats. Electrodes were chronically implanted into the frontal cortex and the dorsal neck muscle of rats for the electroencephalogram (EEG) and electromyogram (EMG), respectively. EEG and EMG were recorded with an electroencephalograph. SleepSigh ver. 2.0 was used for analysis of the sleep-wake state. Total times of waking, non-rapid eye movement (non-REM) sleep, and rapid eye movement (REM) sleep were measured from 10:30 to 16:30. Galantamine had no significant influence on the sleep pattern. On the other hand, donepezil and memantine showed significant increases in sleep latency and total waking time and a decrease in total non-REM sleep time. Furthermore, memantine decreased total REM sleep time. To investigate the characteristics of non-REM sleep in detail, non-REM sleep was classified as stage 1, 2, or 3 according to the depth of sleep. Different from donepezil and galantamine, memantine significantly decreased stage 1 and increased stage 3 in non-REM sleep. From these findings, it can be concluded that galantamine caused no sleep disturbance, different from donepezil and memantine. PMID- 19305124 TI - Propyl gallate, a strong antioxidant, increases the Ca2+ sensitivity of cardiac myofilament. AB - Ca(2+) sensitizers are cardiotonic agents that directly increase the Ca(2+) sensitivity of cardiac myofilament. To find a novel Ca(2+) sensitizer, we have screened a group of phenolic compounds by examining their effects on the Ca(2+) dependent force generation in cardiac muscle fibers. We found that propyl gallate, a strong antioxidant, increased the Ca(2+) sensitivity of cardiac myofilament in a dose-dependent and reversible manner. The present study indicates that propyl gallate is a novel type of Ca(2+) sensitizer with antioxidant activity, which might be more beneficial for the treatment of congestive heart failure associated with oxidative stress than existing Ca(2+) sensitizers. PMID- 19305125 TI - Long-term, six-dimensional live-cell imaging for the mouse preimplantation embryo that does not affect full-term development. AB - Mammalian preimplantation embryonic development is achieved by tightly coordinated regulation of a great variety of temporal and spatial changes. Therefore, it would be valuable to analyze these events three-dimensionally and dynamically. We have previously developed a live-cell imaging method based on the expression of fluorescent proteins, using mRNA injection and time-lapse florescence microscopy. However, with conventional fluorescent microscopy, three dimensional images could not be obtained due to the thickness of the embryos and the optical problem in which ;out-of focus blur' cannot be eliminated. Moreover, as the repeated exposure of intense excitation light to the cell yields phototoxicity, long-term observation was detrimental to embryonic development. Here, we improved our imaging system to enable six-dimensional live-cell imaging of mouse preimplantation embryos (x, y and z axes, time-lapse, multicolor and multisample). Importantly, by improving the imaging devices and optimizing the conditions for imaging, such as intensity of excitation and time intervals for image acquisition, the procedure itself was not detrimental to full-term development, although it is a prolonged imaging process. For example, live pups were obtained from embryos to which two different wavelengths of excitation (488 and 561 nm) were applied at 7.5-min intervals for about 70 h, and 51 images were acquired in the z axis at each time point; thus, a total of 56,814 fluorescent images were taken. All the pups were healthy, reproductively normal and not transgenic. Thus, this live-cell imaging technology is safe for full-term mouse development. This offers a novel approach for developmental and reproductive research in that it enables both retrospective and prospective analyses of development. It might also be applicable to assessment of embryo quality in fields such as human reproductive technology and production animal research. PMID- 19305126 TI - Mouse androgenetic embryonic stem cells differentiated to multiple cell lineages in three embryonic germ layers in vitro. AB - The embryos of some rodents and primates can precede early development without the process of fertilization; however, they cease to develop after implantation because of restricted expressions of imprinting genes. Asexually developed embryos are classified into parthenote/gynogenote and androgenote by their genomic origins. Embryonic stem cells (ESCs) derived from asexual origins have also been reported. To date, ESCs derived from parthenogenetic embryos (PgESCs) have been established in some species, including humans, and the possibility to be alternative sources for autologous cell transplantation in regenerative medicine has been proposed. However, some developmental characteristics, which might be important for therapeutic applications, such as multiple differentiation capacity and transplantability of the ESCs of androgenetic origin (AgESCs) are uncertain. Here, we induced differentiation of mouse AgESCs and observed derivation of neural cells, cardiomyocytes and hepatocytes in vitro. Following differentiated embryoid body (EB) transplantation in various mouse strains including the strain of origin, we found that the EBs could engraft in theoretically MHC-matched strains. Our results indicate that AgESCs possess at least two important characteristics, multiple differentiation properties in vitro and transplantability after differentiation, and suggest that they can also serve as a source of histocompatible tissues for transplantation. PMID- 19305127 TI - Genital blood flow and endometrial gene expression during the preovulatory period after Prostaglandin F(2alpha)-induced luteolysis in different luteal phases in cows. AB - The aim of this study was to examine the hypothesis that follicular and uterine perfusion as well as endometrial gene expression during the ovulatory period differs after induction of luteolysis during the 1(st) follicular wave compared with the 2(nd) wave or in intact cycle. Nine healthy non-lactating Holstein Friesian cows were examined during three estrous cycles. A prostaglandin F(2alpha) analogue (PGF) was administered randomly either on Day 7 (1(st) wave cycle) or Day 11 (2(nd) wave cycle) after detection of ovulation (=Day 1). No hormonal treatment was used in the intact cycle with spontaneous ovulation. Transrectal Doppler sonography was conducted daily after PGF injection and in the intact cycle beginning on Day 18 of the estrous cycle until ovulation. Follicular blood flow (FBF) was determined by measuring the maximum area of colour pixels on digitalized images of the follicles. Uterine blood flow was quantified by the time-averaged maximum velocity (UTAMV) and pulsatility index (PI) in both uterine arteries. Blood flow measurements were carried out on Days -1 and 0. Endometrial biopsy specimens were taken on Day 1 and analyzed for the gene expressions of estrogen, progesterone, oxytocin and VEGF receptors and eNOS and iNOS using RT PCR. The interval from PGF injection to ovulation was shorter (P<0.05) in 1(st) wave cycles than in 2(nd) wave cycles. On Days 0 and -1, FBF was greater (P<0.05) in 1(st) wave cycles than in intact and 2(nd) wave cycles. On Day -1, UTAMV was greater (P<0.05) in 1(st) wave cycles than in intact and 2(nd) wave cycles. There were no differences (P>0.05) in FBF and UTAMV values between 2(nd) wave and intact cycles. No differences (P>0.05) were detected in the gene expressions of endometrial receptors and enzymes between intact, 1(st) and 2(nd) wave cycles. The results show that follicular and uterine perfusion during the ovulatory phase are higher after induction of luteolysis during the 1(st) follicular wave compared with the 2(nd) wave or intact cycle. There were no effects on endometrial gene expression. PMID- 19305128 TI - Effects of N, N-dimethylglycine on the development of in vitro produced bovine embryos. AB - This study investigated the effects of N, N-Dimethylglycine (DMG) on the development of in vitro produced (IVP) bovine embryos. IVP embryos were obtained by in vitro fertilization of in vitro matured oocytes for 6 h. In Experiment 1, IVP embryos were cultured in mSOFaa supplemented with bovine serum albumin but without glucose (SOF1) for 4 days, transferred to mSOFaa (with 5% fetal bovine serum and 1.5 mM glucose; SOF2) supplemented with 0 (control), 0.1,1 or 10 microM DMG and cultured for an additional 7 days (11 days in total) to assess their development in vitro. When cultured in the medium with 0.1 microM DMG, a significantly higher number of IVP embryos developed to the blastocyst and hatched blastocyst stages (40.3 and 40.8%, respectively) compared with the other groups (18.7-31.0% and 15.0-28.7%, respectively; P<0.05, analysis of variance). In Experiment 2, IVP embryos were cultured in SOF1 with or without 0.1 microM DMG for 4 days, transferred to SOF2 with or without 0.1 microM DMG and further cultured as in Experiment 1; DMG was added to either SOF1 or SOF2 and to both of them to assess its exposure effects on embryo development. When cultured continuously with DMG for 11 days, significantly higher rates of IVP embryos developed into blastocyst and hatched blastocyst stages (39.0 and 47.7%, respectively) compared with the other groups (31.0-32.2% and 29.5-31.0%, respectively; P<0.05). In Experiment 3, we examined developmental speed of IVP embryos cultured with or without addition of 0.1 microM DMG to IVC medium after 7 days of IVC. When DMG was added to IVC medium, the ratio of embryos developed to advanced developmental stages (No. of embryos developed to the blastocyst and expanded blastocyst stages/No. of embryos developed to the morula stage) was 28.7% (86/3) and 7 times higher than that of those cultured without DMG, 4.0% (52/13). These results suggest that addition of 0.1 microM DMG to mSOFaa during IVC of IVP bovine embryos has a promoting effect on their development. PMID- 19305129 TI - The centrosomal FOP protein is required for cell cycle progression and survival. AB - FOP is a centrosomal protein originally discovered as a fusion partner of FGFR1 in patients with a rare stem cell myeloproliferative disorder. In DT40 chicken lymphocytes, we show that the normal FOP protein localizes at the centrosome throughout the cell cycle and preferentially accumulates at the distal end of the mother centriole. We used homologous recombination in DT40 cells to generate an inducible null mutant for FOP. Loss of FOP induces apoptosis in the G(1) phase of the cell cycle with accumulation of a 32 kDa P53 tumor suppressor isoform and NOXA and FAS transcripts. However, centrosome integrity and microtubule organization are conserved without FOP and mitotic division and cytokinesis are as efficient as in control cells. Our results suggest that FOP is involved in G(1) to S signaling and thus in proliferation/death fate. Several reports show that centrosome alteration can lead to an arrest in G(1) and, possibly, to senescence in a fraction of cells. The phenotype we observed is more severe in FOP null cells. This could be dependent on the cell context or on the efficiency of a knock out that allows the complete disappearance of the target protein and prevents any de novo synthesis. This is an important observation in regard to the current discussion of what consequence centrosome perturbation could have on a cell and shows that a centrosomal protein can be necessary for cell cycle progression and survival. PMID- 19305131 TI - Camptothecin releases P-TEFb from the inactive 7SK snRNP complex. AB - An immediate effect of DNA Topoisomerase I inhibitors camptothecin (CPT) and its derivates is the inhibition of transcription. These fast-acting drugs are believed to inhibit transcription by blocking topoisomerase-mediated relief of DNA supercoiling that occurs during transcription elongation. The CPT effects are commonly considered to be due to a collision between the drug-trapped enzyme on the DNA template and the elongating RNAPII. Here we present evidences that CPT treatment induces an early affect on the positive elongation factor b (P-TEFb). The P-TEFb activity is tightly and dynamically regulated, and a reservoir of P TEFb is kept in an inactive state in the multisubunit 7SK snRNP. We found that, shortly after treatment, CPT disrupts the large inactive P-TEFB complex, and such effect is reversible and independent from DNA replication. Thus, CPT modulates P TEFb equilibrium in a manner similar to Flavopiridol (FP), a pan-Cdk inhibitor proposed as chemotherapeutic agents against cancers. We determined that while FP inhibits Cdk9 leading to hypo-phosphorylation of RNA polymerase II, CPT-mediated release of free P-TEFb correlates with a concomitant hyper-phosphorylation of RNAPII, which in turn alters the levels and distribution of the RNAPII along transcribed genes. The findings that CPT affects P-TEFb activity provide a direct evidence of the mechanism of this drug to inhibit transcription. PMID- 19305132 TI - A novel role for the Drosophila epsin (lqf): involvement in autophagy. AB - Screening P-element-induced mutant collections, 52 lines were selected as potentially defected ones in endocytosis or autophagy. After excluding those which were rescued by 20-hydroxyecdysone treatment, the exact position of the inserted P-element was determined in the remaining lines. In the case of l(3)S011027 stock, the liquid facets (lqf) gene was affected which codes an epsin homolog protein in Drosophila. We reveal that Lqf is essential to the receptor mediated endocytosis of larval serum proteins (LSPs) in the larval fat body cells of Drosophila. In l(3)S011027 line, lack of Lqf fails the formation of autophagosomes thus leading to the arrest of destroying of trophocytes. Transgenic larvae carrying Lqf-RNAi construct were unable to generate endocytic and autophagic vacuoles and led to a prolonged larval stage. On the other hand, Lqf protein showed an exclusive colocalization with the LysoTracker Red- or GFP Atg8a labeled autophagosomes. By using the antiserum generated against the fifth exon of lqf, we demonstrated that prior to the onset of developmental autophagy the Lqf protein was present in the nucleus of fat body cell, but thereafter the protein was localized in the territory of endocytic and autophagic vacuoles. The fact that the inhibition of the target of rapamycin (TOR) did not restore the autophagic process and the normal development in the case of lqf mutant larvae points to that the Lqf is downstream to the TOR, the central kinase of the autophagy pathway. PMID- 19305133 TI - A molecular mechanism of chronological aging in yeast. AB - The molecular mechanisms that cause organismal aging are a topic of intense scrutiny and debate. Dietary restriction extends the life span of many organisms, including yeast, and efforts are underway to understand the biochemical and genetic pathways that regulate this life span extension in model organisms. Here we describe the mechanism by which dietary restriction extends yeast chronological life span, defined as the length of time stationary yeast cells remain viable in a quiescent state. We find that aging under standard culture conditions is the result of a cell-extrinsic component that is linked to the pH of the culture medium. We identify acetic acid as a cell-extrinsic mediator of cell death during chronological aging, and demonstrate that dietary restriction, growth in a non-fermentable carbon source, or transferring cells to water increases chronological life span by reducing or eliminating extracellular acetic acid. Other life span extending environmental and genetic interventions, such as growth in high osmolarity media, deletion of SCH9 or RAS2, increase cellular resistance to acetic acid. We conclude that acetic acid induced mortality is the primary mechanism of chronological aging in yeast under standard conditions. PMID- 19305134 TI - Regulation of MEF2s by chaperone-mediated autophagy. PMID- 19305135 TI - RNA polymerase plays both sides: vivid and bidirectional transcription around and upstream of active promoters. PMID- 19305136 TI - G1 to S transition and pluripotency: two sides of the same coin? PMID- 19305137 TI - High affinity interaction of the p53 peptide-analogue with human Mdm2 and Mdmx. AB - The Mdm2 and Mdmx proteins are the principal negative regulators of the p53 tumor suppressor. Reactivation of p53 activity by disrupting the Mdm2/Mdmx-p53 interactions offers new possibilities for anticancer therapeutics. Here, we present crystal structures of two complexes, a p53-like mutant peptide with the N terminal domains of Mdm2 and Mdmx, respectively. The structures reveal that the p53 mutant peptide (amino acid sequence: LTFEHYWAQLTS) assumes virtually identical conformations in both complexes despite the different shapes of the p53 binding pockets in these two proteins, has a more extended helical nature compared to the Mdm2-bound wild-type p53 peptide, and does not disturb the native folds of Mdm2 or Mdmx. The extension of the helical structure in the mutant p53 peptide greatly improves its binding to Mdm2 and Mdmx. The fluorescence polarization assay that we have developed using this peptide indicates the affinities towards Mdm2 of 3.6 nM and for Mdmx of 6.1 nM, compared to the low micromolar binding of a similar length wild-type p53 peptide to Mdm2/Mdmx. Our assay does not require expensive non-native amino acids, and allows measurements of the interaction with both Mdm2 and Mdmx in identical conditions-without modification of experimental conditions or setups between the two proteins. The structural information presented here, coupled with the robust fluorescence polarization assay, should enable development of a simple pharmacophore model of cross-selective Mdm2-Mdmx/p53 inhibitors. PMID- 19305138 TI - Sgt1 plays polo during cell division. PMID- 19305139 TI - Autophagy, an Achilles' heel AKTing against cancer? AB - Akt has emerged as an attractive cancer therapeutic target with a central role in cell survival, growth, proliferation and metabolism.A key to the clinical success of Akt inhibitors is the maximal possible antitumor efficacy achievable without intolerable side effects. In our recent work, we show that although Akt inhibition does not always induce a clear apoptotic response, autophagy is a more readily detectable response to pan-Akt knockdown or selective small molecule inhibitors of the PI3K/Akt pathway. Autophagy isa catabolic process of bulk lysosomal degradation and recycling of cytoplasmic material and organelles, which can provide a temporary survival mechanism for cells under stress conditions, but can also make cells vulnerable to several forms of cell death under specific circumstances. We hypothesize that autophagy induced by Akt inhibition may sensitize tumor cells to agents targeting the later steps of this lysosomal degradation process. Indeed, agents that interfere with the lysosomal degradation function could precipitate cell death when combined with Akt inhibition and promote complete tumor remissions in preclinical models. These findings suggest that manipulating the autophagic response may be a promising strategy to increase the therapeutic efficacy of Akt inhibitors. PMID- 19305140 TI - Hybrids of aneuploid human cancer cells permit complementation of simple and complex cancer defects. AB - Causes for the complex phenotypes of cancers, such as altered differentiation, invasion and metastasis, are not known, and multigenic defects are likely. In contrast, well-defined deficiencies, such as those affecting DNA-repair mechanisms and enzymatic pathways, are simple, typically caused by one or a few gene mutations. Complementation by introducing defined genetic elements is used to study simple cancer phenotypes, while complementation by the fusion of whole cells is employed occasionally for complex ones. Hybrids formed solely from the common lines (aneuploid due to chromosomal instability, CIN) are rarely reported. We created stable hybrids of two CIN lines, producing a nearly complete genetic sum of the parental karyotypes. Complementation of a simple cancer phenotype, a Fanconi anemia pathway defective in both parental lines, occurred in all hybrids, restoring the normal drug-resistance phenotype. The grossly defective mitotic spindle checkpoint present in both parental lines was partially corrected in some hybrids, supporting a multigenic origin rather than a single gene defect. Using Affymetrix 100K SNP chips, we mapped chromosomal loci differing among the phenotypically distinct hybrid clones. Fusing CIN cell lines to form mapped hybrids offers new tools for positional cloning or classification of simple and complex cancer phenotypes, including mechanical defects and altered drug responses. PMID- 19305141 TI - Grape seed extract protects IEC-6 cells from chemotherapy-induced cytotoxicity and improves parameters of small intestinal mucositis in rats with experimentally induced mucositis. AB - Mucositis is a common side-effect of high-dose chemotherapy regimens. Grape seed extract (GSE) represents a rich source of proanthocyanidins with the potential to decrease oxidative damage and inflammation within the gastrointestinal tract. We evaluated GSE for its capacity to decrease the severity of chemotherapy-induced mucositis in vitro and in vivo. In vitro: GSE was administered to IEC-6 intestinal epithelial cells prior to damage induced by 5-Fluorouracil (5-FU). Cell viability was determined by neutral red assay. In vivo: Female Dark Agouti rats (130-180 g) were gavaged with 1 ml GSE (400 mg/kg) daily (day 3-11) and received 5-FU (150 mg/kg) by intraperitoneal (i.p.) injection on day nine to induce mucositis. Rats were sacrificed at day 12 and intestinal tissues collected for myeloperoxidase and sucrase activity assays and histological analyses. Statistical analysis was performed by one-way ANOVA. GSE prevented the decrease in IEC-6 cell viability induced by 5-FU (p < 0.01). Compared with 5-FU controls, GSE significantly reduced myeloperoxidase activity by 86% and 27% in the proximal jejunum (p < 0.001) and distal ileum (p < 0.05) respectively; decreased qualitative histological scores of damage (p < 0.05) in the proximal jejunum; increased villus height in the proximal jejunum (17%; p < 0.05) and distal ileum (50%; p < 0.01), and attenuated the 5-FU-induced reduction of mucosal thickness by 16% in the jejunum (p < 0.05) and 45% in the ileum (p < 0.01). GSE partially protected IEC-6 cells from 5-FU-induced cytotoxicity and ameliorated intestinal damage induced by 5-FU in rats. GSE may represent a promising prophylactic adjunct to conventional chemotherapy for preventing intestinal mucositis. PMID- 19305142 TI - Prevention of ischemic brain injury by treatment with the membrane penetrating apoptosis inhibitor, TAT-BH4. AB - In acute thromboembolic stroke, neurological damage is due to ischemia-induced apoptotic death of neuronal cells and the surrounding vascular network. Here, we demonstrate that the BH4 domain of the anti-apoptotic protein, Bcl-x(L), attached to the membrane transport peptide, TAT, reduces stroke injury after intracerebroventricular infusion into immature rats subjected to carotid artery ligation and additional exposure to hypoxia. The injected TAT-BH4 entered neuron bodies, maintained brain architecture, protected neuronal and endothelial cells from apoptosis and promoted neuronal stem cell recruitment. In vitro, TAT-BH4 enhanced the survival of endothelial cells exposed to H(2)O(2), increased neuronal differentiation, and induced axonal remodelling of adult neuronal stem cells. These findings indicate that TAT-BH4 administration protects against acute hypoxia/ischemia injury in the brain by preventing endothelial and neuron cell apoptosis and by inducing neuronal plasticity. PMID- 19305143 TI - Stem cells by the shore: Meeting summary of the 2008 MDI Stem Cell Symposium. PMID- 19305144 TI - Upregulation of the CDC25A phosphatase down-stream of the NPM/ALK oncogene participates to anaplastic large cell lymphoma enhanced proliferation. AB - Here, we demonstrate that the expression of the dual specificity phosphatase CDC25A, a key regulator of cell cycle progression, is deregulated in Ba/F3 cells expressing the oncogenic protein NPM/ALK and in human cell lines derived from NPM/ALK-positive anaplastic large cell lymphomas (ALCL). Both transcriptional and post-translational mechanisms account for the constitutive expression of the protein, and the PI3K/Akt pathway is essential for this process. Importantly, pharmacological inhibition of CDC25 dramatically inhibits the proliferation of NPM/ALK-expressing cells, while moderately affecting the proliferation of control Ba/F3 cells. RNA interference-mediated downregulation of CDC25A confirmed that NPM/ALK-expressing cells are highly dependent on this protein for their proliferation. Moreover, similar PI3K/AKt-mediated constitutive expression of CDC25A takes place down-stream of other hematological oncogenes, including BCR/ABL in Chronic Myeloid Leukemia and FLT3-ITD in Acute Myeloid Leukemia. Altogether, our data point to the functional link between hematopoietic oncogenic tyrosine kinases and the G(1) cell cycle regulator CDC25A, and we propose that this protein may be a potential therapeutic target in ALCL and other hematological malignancies. PMID- 19305145 TI - Safety of a refrigerator-stable varicella vaccine (VARIVAX) in healthy 12- to 15 month-old children: A randomized, double-blind, cross-over study. AB - The safety of a single injection of the refrigerator-stable formulation of varicella vaccine VARIVAX was assessed in a blind, randomized, cross-over trial. Five hundred seven healthy children aged 12 to 15 months received subcutaneous injections of VARIVAX on day 0 and the measles, mumps and rubella vaccine (M-M-R II) on day 42 or M-M-R II on day 0 and VARIVAX on day 42. To maintain blinding, injections were given by a study nurse not involved in safety assessments. M-M-R II acted as a reference to validate the safety assessment, as its safety profile is well known in this age range. Parents or legal guardians recorded adverse events for 42 days following each injection. Solicited injection-site reactions (erythema, swelling, pain) were recorded on days 0 to 4. Other injection-site reactions, daily temperature, rashes and systemic adverse events were recorded on days 0 to 42, and serious adverse events until the final study visit. The safety profile of M-M-R II was consistent with previous reports. Following VARIVAX administration, 47.7% of children had at least one vaccine-related adverse event. Solicited injection-site reactions were reported in 13.0% of children, and 17.2% had at least one other injection-site reaction between days 0 and 42. Most reactions were small ( 0.05) in the occurrence of bacteraemia were evident across all groups. Live TH-4 partially normalised mitotic count and histological severity score in 5-FU treated rats. The inhibitory effect of live TH-4 and TH-4 supernatant on crypt fission suggests therapeutic utility in the prevention of disorders characterised by increased crypt fission, such as colorectal carcinoma. PMID- 19305161 TI - Low molecular weight cyclin E is specific in breast cancer and is associated with mechanisms of tumor progression. AB - Low molecular weight (LMW) isoforms of cyclin E are post-translationally generated in breast cancer cells and are associated with aggressive disease and poor prognosis. In this study, the specificity of LMW cyclin E to cancer cells was determined by measuring cyclin E expression in tumor and non-tumor tissue from 340 breast cancer patients. Our results reveal the LMW isoforms were detected significantly more frequently in breast tumor tissue than in adjacent non-tumor breast tissues (p < 0.0001). The biologic consequences of the LMW isoforms were studied using a non-tumorigenic mammary epithelial cell line transfected with the cyclin E isoforms and resulted in increased clonogenicity, the inability to enter quiescence in response to growth factor deprivation and genomic instability compared to the full-length cyclin E. Biochemical differences between the full-length and the LMW isoforms were also evident. Biacore analyses show that the LMW isoforms have more efficient binding to CDK2 compared to full length cyclin E, which could account for the unique biologic consequences observed with the expression of LMW cyclin E. The LMW isoforms of cyclin E are tumor specific, and are biochemically and biologically distinct from the full length cyclin E which could provide a novel role in breast cancer progression. PMID- 19305162 TI - p53 mediates mitochondria dysfunction-triggered autophagy activation and cell death in rat striatum. AB - In vivo administration of the mitochondrial inhibitor 3-nitropropionic acid (3 NP) produces striatal pathology mimicking Huntington disease (HD). However, the mechanisms of cell death induced by metabolic impairment are not fully understood. The present study investigated contributions of p53 signaling pathway to autophagy activation and cell death induced by 3-NP. Rat striatum was intoxicated with 3-NP by stereotaxic injection. Morphological and biochemical analyses demonstrated activation of autophagy in striatal cells as evidenced by increased formation of autophagosomes, the expression of active lysosomal cathepsin B and D, microtubule associate protein light chain 3 (LC3) and conversion of LC3-I to LC3-II. 3-NP upregulated the expression of tumor suppressor protein 53 (p53) and its target genes including Bax, p53-upregulated modulator of apoptosis (PUMA) and damage-regulated autophagy modulator (DRAM). 3 NP-induced elevations in pro-apoptotic proteins Bax and PUMA, autophagic proteins LC3-II and DRAM were significantly reduced by the p53 specific inhibitor pifithrin-alpha (PFT). PFT also significantly inhibited 3-NP-induced striatal damage. Similarly, 3-NP-induced DNA fragmentation and striatal cell death were robustly attenuated by the autophagy inhibitor 3-methyladenine (3-MA) and bafilomycin A1 (BFA). These results suggest that p53 plays roles in signaling both autophagy and apoptosis. Autophagy, at least partially, contributes to neurodegeneration induced by mitochondria dysfunction. PMID- 19305163 TI - Potent antitumor efficacy of interleukin-18 delivered by conditionally replicative adenovirus vector in renal cell carcinoma-bearing nude mice via inhibition of angiogenesis. AB - It has been demonstrated that interleukin 18 (IL-18) exerts antitumor activity. In this study, we investigated whether oncolytic adenovirus-mediated gene transfer of IL-18 could induce strong antitumor activity. A tumor-selective replicating adenovirus expressing IL-18 (ZD55-IL-18) was constructed by insertion of an IL-18 expression cassette into the ZD55 vector, which is based on deletion of the adenoviral E1B 55-kDa gene. ZD55-IL-18 could express substantially more IL 18 than Ad-IL-18 because of replication of the vector. It has been shown that ZD55-IL-18 exerted a strong cytopathic effect and significant apoptosis in renal cell carcinoma. ZD55-IL-18 significantly decreased VEGF and CD34 expression in the tumor cells. Treatment of established tumors with ZD55-IL-18 showed much stronger antitumor activity than that induced by ZD55-EGFP or Ad-IL-18. These data indicated that oncolytic adenovirus expressing IL-18 could exert potential antitumor activity via inhibition of angiogenesis and offer a novel approach to cancer therapy. PMID- 19305164 TI - Non-coding RNAs, epigenetic memory and the passage of information to progeny. AB - Over the last few years it has become increasingly apparent that RNA is involved in various forms of gene regulation. While much emphasis has been placed on the role of small non-coding RNAs in post-transcriptional modes of gene regulation it has become apparent that a far more complex scenario exists. Recent observations insinuate a paradigm whereby non-coding RNAs are operative effector molecules in the transcriptional regulation of endogenous gene expression. These observations support a route for how epigenetic gene silencing is directed, maintained and passed on as epigenetic memory in human cells. This perspective will highlight the endogenous effector RNAs and mechanism of action whereby non-coding RNAs transcriptionally regulate gene expression in human cells and discuss these recent observations in the context of human evolution. PMID- 19305165 TI - MEK and EGFR inhibition demonstrate synergistic activity in EGFR-dependent NSCLC. AB - Epidermal growth factor receptor (EGFR) inhibitors are highly effective in treating non-small cell lung cancers (NSCLC) expressing activated EGFR, particularly those harboring EGFR mutations. However, most patients who benefit from EGFR inhibitors achieve only partial responses or stable disease, facilitating the emergence of resistance. Thus, progression-free survival advantages in responding patients are modest. Combination therapy, preferably using agents with synergistic activity, could both improve responses and reduce acquired resistance rates. We hypothesized that combining MEK inhibitors with EGFR inhibitors could result in such a benefit. The MAPK pathway lies downstream of EGFR and transduces both proliferative and survival signals in a variety of cancer types. Inhibitors of this pathway are currently in clinical trials, but little evidence exists supporting the use of these agents as monotherapy in EGFR dependent non-small cell lung cancer. In this study, we find EGFR-dependent NSCLC cell lines are moderately sensitive to loss of ERK1/2 activity, either by small molecule inhibition or by siRNA knockdown. The consequence of inhibition is dependent upon the trophic content of the culture media, primarily anti proliferative in serum-rich conditions and pro-apoptotic in serum-poor conditions. However, when ERK inhibition combined with EGFR inhibitors, cytotoxic synergy was observed for all EGFR-dependent cell lines tested in serum-containing media. Enhanced cytotoxicity is demonstrated in cell lines with and without EGFR mutations, including those harboring the T790M escape mutation. These findings support future clinical studies that combine EGFR- and MEK1/2-targeted agents to investigate whether improved outcomes can be achieved in clinically screened EGFR dependent NSCLC. PMID- 19305166 TI - Cdc14p regulates condensin binding to rDNA. PMID- 19305167 TI - Cyclin E as a prognostic factor: what is the question? PMID- 19305185 TI - Reliability and reproducibility of clinic and home blood pressure measurements in hypertensive women according to age and ethnicity. AB - BACKGROUND: The purposes of this study were to establish the reproducibility and reliability of clinic and home blood pressure readings and to determine whether correlations differed according to age and ethnicity. METHODS: Blood pressure readings taken in a clinical setting and at home from 161 hypertensive women who were either younger or older (including 91 White American and 61 African American) were compared with 24-h ambulatory blood pressure monitoring (ABPM) readings (considered the gold standard of blood pressure measurement). RESULTS: Bland-Altman statistical method showed good levels of agreement between clinic blood pressures measured 30 days apart, and blood pressures measured at home in the morning over a 30-day program, when compared with mean 24-h ABPM readings. On examining individual Bland-Altman plots for younger and older women, White American and African-American women's blood pressures were well correlated for home measures and 24-h ABPM readings. The correlation between daytime systolic home blood pressure readings and systolic 24-h ABPM readings was much stronger for White American women (r=0.75) than for African-American women (r=0.57). There were also correlation differences in mean systolic blood pressure between home blood pressure readings and 24-h ABPM readings according to age (r=0.66 for younger and r=0.72 for older). CONCLUSION: These results support current research findings that home blood pressure measurements are reliable when compared with 24 h ABPM readings both in African-American and White women. PMID- 19305186 TI - Improvement of aortic reflection wave responses 6 months after stopping smoking: a prospective study. AB - BACKGROUND: Cigarette smoking is one important preventable cause of cardiovascular illness that has been associated with increased stiffness of large arteries and wave reflection, which are independent predictors of cardiovascular disease. METHODS: We investigated the effect of a 6-month quitting smoking programme on aortic stiffness, central pressure and wave reflections in chronic heavy smokers. Seventy-one (83% normotensives) long-term heavy smokers [>20 cigarettes/day and exhaled carboxy haemoglobin (COHb) (> or =4% and CO ppm > or =21)] completed a 6-month psychological-based stop-smoking program. Patients were divided into two groups. Thirty-one patients aged 45+/-2 years, 71% male, fully quit smoking for 6 months (COHb <2% and CO ppm <10 at 1, 3 and 6 months)--group I, whereas 40 patients aged 45+/-1 years, 73% male, did not change their smoking habits--group II. We measured between baseline and changes after 6 months in aortic stiffness assessed as pulse wave velocity (Complior), central-peripheral pulse pressure (PP) amplification ratio (PPAr), wave reflection (augmentation index corrected for heart rate), augmentation pressure and transit time (Sphygmocor). Ambulatory 24 h blood pressure (ABP) data were obtained at baseline and after 6 months in 36 patients (n = 19) of group I and 17 of group II. RESULTS: In group I, but not in group II, there were significant reductions of peripheral systolic BP (baseline: 131+/-4 to after 6 months: 127+/-4 mmHg, P<0.04) and of systolic BP (baseline: 121+/-4 to after 6 months: 114+/-4 mmHg, P<0.03). In group I from baseline to after 6 months, PPAr increased by 13.1+/ 4.1% and transit time (ms) by 8.7+/-2.9%; augmentation index was reduced by 9.2+/ 1.6%, augmentation pressure by 5.7+/-1.9 mmHg (all P<0.03 vs. baseline and vs. group II); and pulse wave velocity by 2.3+/-2.8% (P = 0.062). In group I, only daytime ABP was reduced from baseline to after 6 months (129/82+/-2/2 to 123/78+/ 2/2 mmHg, P<0.03), whereas no change in 24h-ABP was found in group II. CONCLUSION: Six months of smoking cessation is associated with clear improvement of reflected waves, central pressure and aortic stiffness, and with a reduction of daytime BP. This may contribute to the improvement of cardiovascular prognosis attributed to smoking cessation. PMID- 19305187 TI - Digital recording system of sphygmomanometry. AB - OBJECTIVES: Validation of blood pressure measurement devices is always difficult because the gold standard method depends on the observer's manual assessment. To improve the assessment algorithm in automatic blood pressure measurement devices, knowing the reference blood pressure values and analyzing various data are always necessary. A digital system to record Korotkoff sound and cuff pressure is suggested. METHODS: The recording system consists of cuff, microphone, amplifier, and all the components of an oscillometric measurement device, which enable the collection of various data from patients. After collecting the data, the browsing software can playback the recorded sound and pressure variation on a computer. RESULTS: The system records the Korotkoff sound faithfully. The sound can be played with sound card on a personal computer. Cuff pressure can also be displayed with Korotkoff sound at the same time. With this playback software, the observer can assess systolic blood pressure and diastolic blood pressure . The observer can listen again and discuss with others when the decision is not clear. CONCLUSION: The digital recording system of sphygmomanometry can be used for validation of blood pressure measurement devices, collecting of oscillometric data for research, and educating the students who learn blood pressure measurement. PMID- 19305188 TI - The Omron Elite 7300W home blood pressure monitor passes the European Society of Hypertension International Validation Protocol for women and men. AB - OBJECTIVE: To test the accuracy of the Omron Elite 7300W home blood pressure monitor using the European Society of Hypertension (ESH) Protocol. METHODS: The device was tested using the European Society of Hypertension International Protocol in 33 women and then in 10 men to satisfy the ESH for both men and women. RESULTS: The average error in all participants was 1.2+/-5.2 (SD) [(range 10 to 17)/3.29+/-5.5 (-6 to 17)] mmHg. This is the first device to have been tested using the ESH protocol for the special population of women only. The additional testing of men in the standard ESH protocol enables the validation of this device specifically in women but also in men. However, as with all automatic devices, it is important to validate its accuracy in each individual patient who uses it and to monitor its accuracy intermittently in each individual patient. CONCLUSION: The Omron Elite 7300W home blood pressure monitor passed the ESH validation protocol and can be recommended for patient use. PMID- 19305189 TI - Accuracy of automated oscillometric blood pressure monitors in the detection of cardiac arrhythmias. PMID- 19305190 TI - Oculoauriculovertebral spectrum: report of nine familial cases with evidence of autosomal dominant inheritance and review of the literature. AB - Oculoauriculovertebral spectrum (OAVS; OMIM 164210) is a complex condition characterized by defects of aural, oral, mandibular and vertebral development. The aetiology of this condition is likely to be heterogeneous; most cases are sporadic, however, familial cases suggesting autosomal recessive and autosomal dominant inheritance have been reported. In this study, we describe the clinical aspects of nine familial cases with evidence of autosomal dominant inheritance and compare them with reports in the literature. Interfamilial and intrafamilial clinical variabilities were observed in this study (reinforcing the necessity of careful examination of familial members). We suggest that oculoauriculovertebral spectrum with autosomal dominant inheritance is characterized mainly by bilateral auricular involvement and rarely presents extracranial anomalies. PMID- 19305191 TI - Hypertrichosis, hyperkeratosis and mental retardation syndrome: further delineation of phenotype. PMID- 19305192 TI - Milder form of pachydermoperiostosis: a report of four cases. AB - Pachydermoperiostosis is known to occur in three distinct forms: complete, incomplete, and form fruste. We report four patients with clubbing and a cylindrical appearance of lower legs without pachydermia and periostosis. We believe that this represents a milder, but distinct subgroup of pachydermoperiostosis. One family showed probable autosomal recessive inheritance. PMID- 19305193 TI - Oromandibular limb hypogenesis syndrome with no oromandibular features, or Moebius syndrome without facial palsy? A diagnostic conundrum. PMID- 19305194 TI - Nitric oxide and the stress response--two 2-edged swords swung together. PMID- 19305195 TI - Beyond the bout--new perspectives on exercise and fat oxidation. PMID- 19305196 TI - Potential benefits of warm-up for neuromuscular performance of older athletes. AB - Demographic trends project increasing numbers of older people to engage in exercise programs and sports. Sustained participation depends on both perceived health outcomes and avoidance of debilitating injuries. This review explores the potential benefits of physiologically based warm-up strategies to alleviate some key age-related decreases in the biomechanical capacity for skilled sport movements. PMID- 19305197 TI - Plasticity of the human tendon to short- and long-term mechanical loading. AB - This review examines the effects of short- and long-term static and cyclic mechanical loading on the mechanical properties of tendons. Tendons do not alter their mechanical properties after static and cycling loading that occurs during fatiguing contractions. Adaptations can occur after the application of long-term loading, but the strain magnitude must exceed a given threshold. PMID- 19305198 TI - Exercise regulates heat shock proteins and nitric oxide. AB - Heat shock proteins (HSP) produced during exercise have important roles in modulating the effects of oxidative stresses to preserve cellular function. The ability of HSP to regulate nitric oxide, which has both protective and detrimental effects on the airways and circulation, may have implications for exercise-induced asthma. Exercise regulates HSP and nitric oxide interactions and protects against organ dysfunction and oxidative stresses. PMID- 19305199 TI - Blood flow restricted exercise and skeletal muscle health. AB - For nearly half a century, high mechanical loading and mechanotransduction pathways have guided exercise recommendations for inducing muscle hypertrophy. However, emerging research on low-intensity exercise with blood flow restriction challenges this paradigm. This article will describe the BFR exercise model and discuss its efficacy, potential mechanisms, and clinical viability. PMID- 19305200 TI - Basal and exercise-induced regulation of skeletal muscle capillarization. AB - Regulation of skeletal muscle capillarization involves distinct signaling pathways and growth factors including nitric oxide and vascular endothelial growth factor. Our understanding of this complex regulation continues to expand with the identification of new angiogenic growth factors. Future work needs to increase the use of advanced molecular techniques to expand our knowledge of the regulation of basal and exercise-induced capillarization. PMID- 19305201 TI - Exercise improves fat metabolism in muscle but does not increase 24-h fat oxidation. AB - Despite decades of research into the effects of exercise on fat metabolism, there is still no clear understanding of how exercise helps to regulate fat mass. Although exercise improves the capacity of muscle to oxidize fat, our studies suggest that moderate duration exercise (< or =1 h) has little impact on 24-h fat oxidation. PMID- 19305202 TI - The quadrupedal nature of human bipedal locomotion. AB - During rhythmic movement, arm activity contributes to the neural excitation of leg muscles. These observations are consistent with the emergence of human bipedalism and nonhuman primate arboreal quadrupedal walking. These neural and biomechanical linkages could be exploited in rehabilitation after neurotrauma to allow the arms to give the legs a helping hand during gait rehabilitation. PMID- 19305203 TI - How should objectively measured physical activity data be used analytically? PMID- 19305205 TI - Unintentional injuries. PMID- 19305206 TI - Framing the problem: injuries and public health. AB - OBJECTIVES: To introduce the field of injury control and public health approaches to injury prevention. METHODS: A review of injury epidemiology, definitions, intervention approaches, and the importance of injury as a public health problem. RESULTS: Injuries are a large national and international problem affecting families and communities. Injuries are predictable and preventable. Behavioral, environmental, and technological solutions will be necessary to reduce or eliminate injuries. CONCLUSIONS: Reductions in injury and their costs to families and communities are possible but will need support, collaboration, and partnering at the local level. PMID- 19305207 TI - Outdoor recreational injuries: a partnership in prevention. PMID- 19305208 TI - Partnering with mutual aid associations to provide home injury prevention. AB - Injury is the leading cause of death and a leading cause of morbidity among children in the US. Poor, minority children, especially those with language and cultural differences, are disproportionately affected. The traditional childhood injury prevention approaches (eg, physician office counseling, public service announcements) may not be effective when significant language and cultural differences exist between provider and client. Partnering with mutual aid associations to create low literacy education tools for their home visitors to use to teach immigrant families about home safety may be one method to provide injury prevention information. PMID- 19305209 TI - Accountability and assessment of emergency drill performance at schools. AB - Many schools throughout the United States are mandated to hold drills, or operational exercises, to prepare for fires, earthquakes, violence, and other emergencies. However, drills have not been assessed for their effectiveness in improving preparedness at schools. This mixed-methods study measures the quantity and the quality of drills in an urban school district in Los Angeles. Compliance with California mandates was fair; most schools barely met requirements. Drills were not used as opportunities to improve procedures. Sites neither conducted any self-assessments nor made changes to procedures on the basis of performance. Suggestions include developing realistic simulated exercises, debriefing, and better school accountability for drills. PMID- 19305210 TI - Injury prevention counseling for adults: have we made progress? AB - Injuries are a leading cause of morbidity and mortality in the United States. Physicians are a trusted source that can disseminate information about the prevention of injury and violence. This study used the second Injury Control and Risk Survey to report the national prevalence of healthcare provider injury prevention counseling to adults. Results indicate that overall 1 in 5 adults who visited a healthcare provider received some counseling on injury prevention. Counseling prevalence varied by injury topic and patient demographic characteristics. Many people who could benefit from counseling are not receiving it, even among those who had visited a healthcare provider. PMID- 19305211 TI - Parent/Caregiver supervision and child injury: a systematic review of critical dimensions for understanding this relationship. AB - This study reviewed the relationship between recognized dimensions of supervision and children's injuries based on Saluja et al's (Injury Control and Safety Promotion. 2004;11:17-22) hierarchal model of supervision strategies. A systematic review of peer-review studies was done with the earliest records available to 2007. There were 112 potentially relevant articles identified; 31 studies met all inclusion criteria. Reported studies were categorized according to the dimensions of supervision they addressed. Studies were not evenly distributed across the dimensions. There was evidence from the study that directly linking dimensions of supervision to child injury risk and outcomes is scarce. Future studies should consider attention, proximity, and continuity of supervision to provide a holistic understanding of the relationship between supervision and injury. PMID- 19305212 TI - Maternal rules, compliance, and injuries to preschool children. AB - This study examined the relationships among maternal rules, child compliance, and the injury experience of preschool children. To do so, 278 mothers of 3-year-old children were interviewed, mothers and children were videotaped, and medical records were reviewed. Results indicated that mothers' observed rule enforcement and children's committed compliance were inversely related to injury experience. The authors concluded that it is not the number or self-reported insistence on rules that are associated with injuries. Rather, it is only the mother's follow through even in situations in which there was no perceived risk of injury that has an impact. Rules must be enforced and compliance encouraged in a manner that supports the development of autonomously motivated behavior. PMID- 19305213 TI - Parental safety-related knowledge and practices associated with visiting a mobile safety center in a low-income urban population. AB - A mobile safety center (MSC) provided education and reduced-cost safety products to low-income urban families. We evaluated uptake of this service under 3 different conditions, and safety-related knowledge and behavior associated with visiting the MSC among 210 families. Utilization varied widely under the 3 different conditions. At follow-up, MSC visitors scored slightly higher on a knowledge test than nonvisitors and improved more in reported car safety seat use, but did not differ in observed safety product use. This study provides very modest evidence of a positive impact of the MSC when its services are provided at a community health center. PMID- 19305214 TI - Associations between childhood bicycle helmet use, current use, and family and community factors among college students. AB - Bicycle helmet use from elementary school to high school was examined using retrospective reports collected from 166 students at a university in northern Colorado. Bicycle helmet use in elementary school was a common practice, especially among students born after 1980. However, a rapid decline in bicycle helmet use in junior high school and high school was reported. Students with higher perceived norm of family bicycle helmet use wore helmets more often in junior high school and high school than students with a low family norm. Growing up in the bicycle-friendly community was positively associated with consistent bicycle helmet use. PMID- 19305215 TI - Alcohol-impaired driving and children in the household. AB - More children in the United States are killed in motor vehicle crashes annually than by any other cause; nearly a quarter of these deaths involve alcohol. This study examines the national prevalence of alcohol-impaired driving and riding with an alcohol-impaired driver and the association of these behaviors to having at least 1 child in the household. An estimated 2.5 million adult drivers with children living in their households reported that they had been a recent alcohol impaired driver. Evidence-based approaches, including mass media campaigns and sobriety checkpoints, continue to be critically important public health activities. PMID- 19305216 TI - Efficacy of a brief group parent-teen intervention in driver education to reduce teenage driver injury risk: a pilot study. AB - The purpose of this study was to test the efficacy of an adapted Checkpoints Program designed to increase parental limits on novice teen independent driving under high-risk conditions. Twenty-seven class sessions with a minimum of 5 dyads each were delivered in driver education to 231 parent-teen dyads. Entire driving school classes were randomized to Checkpoints Program or comparison group sessions, both led by a trained health educator. At licensure, compared with parents in the comparison group, treatment parents had increased awareness of teen driving risk and were more likely to have completed a parent-teen driving agreement and met Checkpoints recommendations for restrictions on teen driving in inclement weather and road types. They were also marginally more likely to have met Checkpoints restrictions on driving with teen passengers. This study indicates that it is feasible to implement the Checkpoints Program in driver education with positive effects on parent management practices. PMID- 19305217 TI - CDC Childhood Injury Report: Patterns of Unintentional Injuries Among 0- to 19 Year Olds in the United States, 2000-2006. PMID- 19305219 TI - Preface. PMID- 19305220 TI - Incremental health system reform policy: Ecuador's law for the provision of free maternity and child care. AB - This study assessed the impact that the Ley de Maternidad Gratuita y Atencion a la Infancia (LMGAI) [Law for the Provision of Free Maternity and Child Care] in Ecuador has had on health services utilization and infant mortality. These outcomes were also examined by socioeconomic status. This retrospective study used demographic and health surveys, ENDEMAIN 1999 and 2004, with multivariate logistic regression to assess the impact post-LMGAI, controlling for mother's socioeconomic status, maternal and birth history, and demographic characteristics. Primary healthcare services utilization outcomes significantly improved post-LMGAI. Neonatal mortality decreased post-LMGAI. Further evaluation is needed as implementation continues to understand the expansion of primary healthcare services in future health system reforms. PMID- 19305221 TI - Governance and the effectiveness of the Buenos Aires public health insurance implementation process. AB - A multiple-case study was conducted to explain the relationship between the governance of the Buenos Aires Public Health Insurance (PHI) implementation process and its effectiveness in 8 municipalities. The heterogeneity in the effectiveness of PHI implementation was explained by the characteristics of the governance implementation process. The local health authorities' position on the PHI (favorable reception, formal acceptance, or rejection) influenced the nature of the articulation of the PHI within the municipal health system. Support for local management teams on the part of the municipal health and social development authorities affected the possibilities of implementing strategies to overcome unfavorable context-related conditions. PMID- 19305222 TI - A multimodal strategy based on pay-per-performance to improve quality of care of family practitioners in Argentina. AB - Pay-for-performance has become increasingly common to complement physician reimbursement. We designed a quality framework to measure family physicians' performance in a managed care setting in Buenos Aires. We aimed to assess the effectiveness of a multimodal intervention based on pay-for-performance, teamwork, continuous education, and audit and feedback to improve quality. After 2 years, a significant improvement was observed in most of the indicators measuring clinical effectiveness and some improvements were observed in other domains. Despite these results, a better performance matrix is needed to capture not only specific conditions but also other aspects like integrating, prioritizing, and personalizing care. PMID- 19305223 TI - Primary care and avoidable hospitalizations: evidence from Brazil. AB - This article provides evidence of the effectiveness of family-based, community oriented primary healthcare programs on the reduction of ambulatory care sensitive hospitalizations in Brazil. Between 1998 and 2002, expansions of the Family Health Program were associated with reductions in hospitalizations for diabetes mellitus and respiratory problems and Community Health Agents Program expansions were associated with reductions in circulatory conditions hospitalizations. Results were significant for only the female population only, suggesting that these programs were more effective in reaching women than men. Program coverage may have contributed to an estimated 126 000 fewer hospitalizations between 1999 and 2002, corresponding to potential savings of 63 million US dollars. PMID- 19305224 TI - Assessing quality across healthcare subsystems in Mexico. AB - Recent healthcare reform efforts in Mexico have focused on the need to improve the efficiency and equity of a fragmented healthcare system. In light of these reform initiatives, there is a need to assess whether healthcare subsystems are effective at providing high-quality healthcare to all Mexicans. Nationally representative household survey data from the 2006 Encuesta Nacional de Salud y Nutricion (National Health and Nutrition Survey) were used to assess perceived healthcare quality across different subsystems. Using a sample of 7234 survey respondents, we found evidence of substantial heterogeneity in healthcare quality assessments across healthcare subsystems favoring private providers over social security institutions. These differences across subsystems remained even after adjusting for socioeconomic, demographic, and health factors. Our analysis suggests that improvements in efficiency and equity can be achieved by assessing the factors that contribute to heterogeneity in quality across subsystems. PMID- 19305225 TI - Local health governance in central Brazil. AB - The study evaluates the performance of local health councils and their capacity to promote accountability and improve primary healthcare in central Brazil. An exploratory descriptive study was carried out with the application of questionnaires to analyze their organization and functioning. Results show that they approve health plans and management of public funds but have little autonomy to define health policies. They function mostly bureaucratically with minimum intersectorial articulation. Their capacity to improve delivery of healthcare is limited, but accountability has improved. Thus, it is necessary to enhance the autonomy of these councils, provide technical qualification, and enhance community participation. PMID- 19305226 TI - A conversation on health in Canada: revisiting universality and the centrality of primary healthcare. AB - In 2006, British Columbia launched a public consultation on how to strengthen the health system. We report on the processes and the inputs and views submitted and examine the perceived importance of universality and primary healthcare (PHC). Public response revealed strong support for the Canada Health Act, which upholds 5 principles: public administration, comprehensiveness, universality, portability, and accessibility, and also a need for the system to be more open to innovation. It recognized that keys to improving population health and efficiency gains within the health system lie within the scope of PHC and that prevention, demand management, and self-management are all part of PHC. PMID- 19305227 TI - The impact of primary healthcare on population health in low- and middle-income countries. AB - This article assesses 36 peer-reviewed studies of the impact of primary healthcare (PHC) on health outcomes in low- and middle-income countries. Studies were abstracted and assessed according to where they took place, the research design used, target population, primary care measures, and overall conclusions. Results indicate that the bulk of evidence for PHC effectiveness is focused on infant and child health, but there is also evidence of the positive role PHC has on population health over time. Although the peer-reviewed literature is lacking in rigorous experimental studies, a small number of relatively well-designed observational studies and the consistency of findings generally support the contention that an integrated approach to primary care can improve health. A few large-scale experiences also help identify elements of good practice. The review concludes with several recommendations for future studies, including a focus on better conceptualizing and measuring PHC, further investigation into the advantages of comprehensive over selective PHC, need for experimental or quasi experimental research designs that allow testing of the independent effect of primary care on outcomes over time, and a more detailed conceptual framework guiding overall evaluation design that places limits on the parameters under consideration and describes relationships among different levels and types of data likely to be collected in the evaluation process. PMID- 19305228 TI - TROT Line: Live and Direct From the Republic of Texas. PMID- 19305229 TI - What if ...: you're starting an IV and the catheter breaks off in the vein? PMID- 19305230 TI - Resistance training in the treatment of diabetes and obesity: mechanisms and outcomes. AB - Resistance exercise training (RET) is gaining broad acceptance as a complement to endurance exercise in the treatment of cardiovascular risk factors. This article reviews the most current and reliable literature regarding the biological mechanisms and potential clinical effectiveness of RET in the treatment of 2 major cardiovascular risk factors, diabetes and obesity, obtained from human subject studies found by querying MEDLINE Plus/Ovid literature search system for the years 1950-2008. RET appears to enhance insulin sensitivity and improve glucose tolerance in a wide range of study groups. In addition, studies have shown that improved glucose uptake is not a mere consequence of the typical increase in fat-free mass associated with RET but is likely a result of qualitative changes in resistance-trained muscle. There is also substantial evidence that regular RET can effectively alter body composition in both men and women. It has been shown to increase total fat-free mass, muscular strength, and resting metabolic rate, and preferentially mobilize the visceral and subcutaneous adipose tissue in the abdominal region. The studies presented in this review demonstrate that RET should remain an important focus of translational research, where clinical trials of RET encourage the performance of mechanistic studies and where mechanistic studies lead to further clinical trials. PMID- 19305231 TI - Clinical manifestations and consequences of obstructive sleep apnea. AB - Obstructive sleep apnea is a common respiratory disorder that is underdiagnosed and associated with a variety of adverse health and safety consequences. Treatment is effective in improving quality of life and reducing morbidity. This underscores the importance of considering the diagnosis in suitable patients, verifying the diagnosis, and initiating prompt, effective therapy. In this review, the risk factors, symptoms and signs, diagnosis, clinical consequences, and treatment of obstructive sleep apnea are discussed. PMID- 19305232 TI - Achievement of heart health characteristics through participation in an intensive lifestyle change program (Coronary Artery Disease Reversal Study). AB - PURPOSE: Lifestyle habits and cardiovascular disease (CVD) risk factors are closely linked. Unfortunately, few individuals meet the goals for cardiovascular health that are recommended in public health initiatives. The purpose of this study was to determine the effect of an intensive lifestyle intervention program on the achievement of a group of recognized heart health characteristics as well as on the reduction of individual CVD risk factors. METHODS: Of 200 military healthcare beneficiaries with coronary artery disease or CVD risk factors (mean age = 61 years) who entered a 1-year, prospective, cohort, multicomponent lifestyle intervention study (lacto-ovo vegetarian diet, exercise, stress management, group support), 186 subjects enrolled and 144 participated for 1 year. RESULTS: At 3 months and 1 year compared with baseline, the proportion of subjects meeting 5 recognized heart health characteristics improved (P < .001): fiber intake >25 g/d (94% and 72% vs 35%); exercise > or =150 min/wk (79% and 58% vs 31%); low-density lipoprotein cholesterol <100 mg/dL (75% and 63% vs 46%); body mass index <25 kg/m (34% and 38% vs 23%); and blood pressure <140/90 mm Hg (84% and 83% vs 69%). At 1 year, more subjects (72% vs 32% at baseline), especially those with intervention adherence above (94%) versus below (58%) the study population median (P < .0005), achieved 3 or more of these characteristics. CONCLUSION: An intensive lifestyle intervention promotes achievement of important heart health characteristics that, if maintained, may substantially reduce CVD events. PMID- 19305234 TI - Cardiac rehabilitation after stroke-need and opportunity. AB - PURPOSE: In North America, established long-term rehabilitation programs are commonly available for individuals following a cardiac event but are largely unavailable for stroke survivors. The purpose of this study was to determine (1) the availability of cardiac rehabilitation (CR) for individuals poststroke (survey of all programs in Ontario, Canada) and (2) the effects of CR, comparing individuals with primary and secondary diagnoses of stroke versus those with cardiac diagnoses only (retrospective review of a large outpatient North American program). METHODS: An Ontario-wide survey was disseminated to CR programs to determine barriers to enrollment of stroke participants. Additionally, a retrospective analysis of data from 9,173 participants in 1 CR program in Toronto, Ontario, compared 3 subgroups (n = 19 each): (1) primary diagnosis of stroke or transient ischemic attack (TIA), (2) primary cardiac diagnosis and occurrence of stroke or TIA, and (3) cardiac diagnoses only. RESULTS: Twenty-four of 40 (60%) programs surveyed included stroke participants, although the proportion was small (<5% of total enrollment). Barriers to enrollment included issues around primary diagnosis and degree of stroke-related disability. While those with a history of stroke or TIA had a lower baseline peak oxygen uptake, all 3 groups showed comparable postprogram improvements in peak oxygen uptake and anaerobic threshold (time effect, P < .001). There were no group-time interaction effects. CONCLUSIONS: Despite the common cardiovascular etiology of stroke and heart disease, individuals with stroke are not routinely included in CR in Ontario. However, individuals with stroke demonstrated similar training-related improvements in exercise capacity compared with nonstroke participants. PMID- 19305235 TI - Effects of a pedometer-based intervention on physical activity levels after cardiac rehabilitation: a randomized controlled trial. AB - PURPOSE: In Australia, patient adherence to physical activity after a cardiac rehabilitation program (CRP) is poor. This study evaluated the efficacy of a pedometer-based intervention to increase physical activity after CRP. METHODS: Patients (n = 110) who had attended a CRP were randomized into an intervention or a control group. The 6-week intervention included self-monitored physical activity using a pedometer and step calendar and 2 behavioral counseling and goal setting sessions. Self-reported physical activity and psychosocial status were collected at baseline, 6 weeks, and 6 months. Participant exercise capacity was measured using a gas exchange analysis system. RESULTS: Study groups were not significantly different at baseline, except for occupation. At 6 weeks and 6 months, improvements in total physical activity sessions (P = .002 and P = .016, respectively), walking minutes (P = .013, 6 weeks only), and walking sessions (P < .001 and P = .035) in the intervention group were significantly greater than those in the control group after adjusting for baseline differences. At 6 months, total physical activity minutes in the intervention group also increased significantly more than those in the control group (P = .044). These self reported behavioral changes were corroborated by improvements in cardiorespiratory fitness at 6 months in the intervention group (P = .01). There were also significant improvements in psychosocial health at 6 weeks and 6 months in the intervention group. CONCLUSION: The pedometer-based intervention was successful in increasing physical activity in cardiac patients after a CRP. This intervention could be given to patients to promote adherence to physical activity guidelines after a CRP, particularly in centers where maintenance programs are not available. PMID- 19305236 TI - Effects of attending phase II cardiac rehabilitation on patient versus spouse (proxy) quality-of-life perceptions. AB - PURPOSE: Little has been published about the differences or similarities in patient and spouse views of the patient's health-related quality of life (HRQL) related to cardiac rehabilitation (CR) participation. This study described patient versus spouse (proxy) ratings of patient HRQL prior to (T1) and after 6 weeks (T2) of phase II CR. METHODS: A nonprobability sample of patients referred to phase II CR and their spouses were recruited (42 pairs). The Short Form 36 v.2 (SF-36) instrument was used to measure HRQL. Differences between group means were tested with paired-samples t tests (P < .05). RESULTS: Improvements in patient self-report of HRQL from T1 to T2 were noted in physical functioning, role physical, bodily pain, vitality, social functioning, role-emotional (all P <.001), mental health (P < .01), and both physical and mental component summary (P < .001). Spouse reports of patient HRQL improved in physical functioning, role physical, bodily pain, vitality, social functioning, mental health (all P < .001), role-emotional (P < .01), and the physical (P < .001) and mental component summary (P < .01). Differences between patient and spouse were found at T1 in vitality and mental health (both P < .01). At T2, differences between patient and spouse were found in vitality (P < .001), physical functioning, mental health (both P < .05), and the mental component summary (P < .05). CONCLUSIONS: Improvement of patient HRQL is mirrored by spouse viewpoint of patient HRQL. This unique finding has not previously been reported in the literature. PMID- 19305237 TI - The role of emotional imagery and somatosensory amplification in atypical chest pain in patients with angina pectoris: a single-case experimental design. AB - BACKGROUND: Atypical chest pain is a common condition that has poor symptomatic, functional, and psychological outcomes. Individuals with abnormal coronary anatomy who experience chest pain present a particular challenge to medical services as they frequently report other chest sensations that are atypical of their condition. Cognitive-behavioral therapy has been found to be effective in the treatment of atypical chest pain, suggesting that anxiety may have a significant etiological role. Normal or minor physical sensations may be interpreted as evidence of serious medical problems, resulting in a vicious circle of anxiety, physiological arousal, and hypervigilance. METHODS: The present study evaluated the role of emotional imagery and somatosensory amplification in the experience of atypical chest pain in a patient with diagnosed angina pectoris. Using an experimental ABACA reversal design, the patient was assisted to engage in 3 types of imagery via the presentation of 3 prerecorded scripts: (A) neutral, (B) angina pectoris, and (C) idiosyncratic emotional imagery. Subjective measures of atypical chest sensation intensity were collected at 5 points during each script presentation. RESULTS: It was found that the patient experienced more intense chest sensations while engaged in the first angina pectoris imagery condition compared to the second presentation of the neutral script. This difference was found to be statistically significant, using an interrupted time series analysis. CONCLUSIONS: The findings of this study support the view that anxiety processes can exacerbate and intensify the experience of atypical chest sensations in patients who have abnormal coronary anatomy. PMID- 19305238 TI - Interval training versus continuous training in patients with chronic obstructive pulmonary disease. AB - PURPOSE: The purpose of this study was to compare the effects of interval training (IT) and continuous steady-pace training (CT) in patients with COPD. METHODS: Patients (n = 21) (mean forced expiratory volume in 1 second ([FEV1] = 44.6% +/- 13.9%) were randomized to IT, and 20 patients (mean FEV1 = 41.7% +/- 12.6%) to CT. Outcome measures included 6-minute walk distance, maximal work capacity, endurance exercise time during constant workload exercise at 60% to 70% of maximal work capacity, and quality of life including fatigue and dyspnea. Participants exercised 3 times per week for 8 weeks, and total work was the same for both training regimens. RESULTS: Significant improvement in mean score was observed in each variable within each of the 2 groups: 6-minute walk distance (IT = 158 +/- 178 ft, CT = 106 +/- 165 ft); maximal work capacity (IT = 10.0 +/- 13.0 W, CT = 11.5 +/- 13.1 W); endurance exercise time (IT = 15.0 +/- 12.5 minutes, CT = 18.7 +/- 10.6 minutes); and quality of life domains, fatigue (IT = 3.1 +/- 3.0, CT = 2.8 +/- 4.7), and dyspnea (IT = 4.4 +/- 5.3, CT = 5.4 +/- 5.1). There was no significant difference in the extent of improvement between the 2 training regimens for any of the outcome variables. CONCLUSION: Compared with CT, IT was well tolerated and produced similar improvements in exercise performance and quality of life. PMID- 19305239 TI - Efficacy and tolerability of yoga breathing in patients with chronic obstructive pulmonary disease: a pilot study. AB - PURPOSE: Yoga-derived breathing has been reported to improve gas exchange in patients with chronic heart failure and in participants exposed to high-altitude hypoxia. We investigated the tolerability and effect of yoga breathing on ventilatory pattern and oxygenation in patients with chronic obstructive pulmonary disease (COPD). METHODS: Patients with COPD (N = 11, 3 women) without previous yoga practice and taking only short-acting beta2-adrenergic blocking drugs were enrolled. Ventilatory pattern and oxygen saturation were monitored by means of inductive plethysmography during 30-minute spontaneous breathing at rest (sb) and during a 30-minute yoga lesson (y). During the yoga lesson, the patients were requested to mobilize in sequence the diaphragm, lower chest, and upper chest adopting a slower and deeper breathing. We evaluated oxygen saturation (SaO2%), tidal volume (VT), minute ventilation (E), respiratory rate (i>f), inspiratory time, total breath time, fractional inspiratory time, an index of thoracoabdominal coordination, and an index of rapid shallow breathing. Changes in dyspnea during the yoga lesson were assessed with the Borg scale. RESULTS: During the yoga lesson, data showed the adoption of a deeper and slower breathing pattern (VTsb L 0.54[0.04], VTy L 0.74[0.08], P = .01; i>fsb 20.8[1.3], i>fy 13.8[0.2], P = .001) and a significant improvement in SaO2% with no change in E (SaO2%sb 91.5%[1.13], SaO2%y 93.5%[0.99], P = .02; Esb L/min 11.2[1.1], Ey L/min 10.2[0.9]). All the participants reported to be comfortable during the yoga lesson, with no increase in dyspnea index. CONCLUSION: We conclude that short term training in yoga is well tolerated and induces favorable respiratory changes in patients with COPD. PMID- 19305240 TI - Infometrics, technometrics, and the power of the digital written word. PMID- 19305242 TI - Craniofacial autologous fat transfer. PMID- 19305243 TI - Perspectives in cell-based skeletal tissue. PMID- 19305244 TI - Computational fluid dynamic study on obstructive sleep apnea syndrome treated with maxillomandibular advancement. AB - Maxillomandibular advancement is one of the treatments available for obstructive sleep apnea. The influence of this surgery on the upper airway and its mechanism are not fully understood. The present research simulates the flow fields of narrowed upper airways of 2 patients with obstructive sleep apnea treated with maxillomandibular advancement. The geometry of the upper airway was reconstructed from computed tomographic images taken before and after surgery. The consequent three-dimensional surface model was rendered for measurement and computational fluid dynamics simulation. Patients showed clinical improvement 6 months after surgery. The cross-sectional area of the narrowest part of the upper airway was increased in all dimensions. The simulated results showed a less constricted upper airway, with less velocity change and a decreased pressure gradient across the whole conduit during passage of air. Less breathing effort is therefore expected to achieve equivalent ventilation with the postoperative airway. This study demonstrates the possibility of computational fluid dynamics in providing information for understanding the pathogenesis of OSA and the effects of its treatment. PMID- 19305245 TI - Magnesium-based bone cement and bone void filler: preliminary experimental studies. AB - Bone cement has great potential in craniofacial surgery in the repair of osseous defects secondary to surgery or trauma. This includes the use of bone cement as a bone void filler for full-thickness cranial defects and as augmentation of deficient bones. Ideally, this material should be easily available, biocompatible, resorbable, bone inductive, and have adhesive qualities to bone. Calcium-based bone cements have some of these qualities but have a higher than desirable failure rate. OsteoCrete, a new magnesium-based bone cement and bone void filler, was compared to Norian in critical-sized skull defects and cementing bone flaps in rabbits. Both materials were successful; however, OsteoCrete had a faster resorption and replacement by bone rate than Norian. Bone flap position and apparent stability were also superior with OsteoCrete. There were no adverse reactions to either cement. A magnesium-based bone cement presents with advantages when compared with a comparator calcium-based cement in craniofacial surgery. PMID- 19305246 TI - Crown morphologic abnormalities in the permanent dentition of patients with cleft lip and palate. AB - The purpose of this study was to investigate crown morphologic abnormalities in the permanent dentition of subjects with clefts and unaffected controls. This is a prospective study of 100 patients and 60 control subjects. The study was carried out at the Cleft Lip and Palate Center at the King Abdullah University Hospital and at the Maxillofacial Unit at Jordan University of Science and Technology during the period November 2003 to September 2005. Eleven kinds of crown morphology abnormalities commonly described in the permanent dentition were recorded using the Arizona State University Dental Anthropology System. There were 1525 teeth examined in 100 subjects with clefts, and 203 crown morphologic abnormalities were recorded, resulting in a mean of 2 abnormalities per individual. No significant difference between the unilateral and bilateral cleft lip and palate groups was found; however, the difference between the cleft group and the control group was highly significant. Crown morphologic abnormalities were more frequent in the maxillary cleft teeth than in the mandibular teeth and in the anterior teeth than in posterior teeth, with a statistically significant difference. The most frequently found abnormalities in subjects with clefts were peg-shaped maxillary incisors and missing hypocone, followed by excess mammelons. In conclusion, there was a significant increase in the frequency of crown morphologic abnormalities in individuals with clefts when compared with subjects without clefts. Crown morphologic abnormalities occurred throughout the entire dentition and did not follow any predictable pattern. PMID- 19305247 TI - Photoelastic analysis of stress distribution in oral rehabilitation. AB - The purpose of this study was to present a literature review about photoelasticity, a laboratory method for evaluation of implants prosthesis behavior. Fixed or removable prostheses function as levers on supporting teeth, allowing forces to cause tooth movement if not carefully planned. Hence, during treatment planning, the dentist must be aware of the biomechanics involved and prevent movement of supporting teeth, decreasing lever-type forces generated by these prosthesis. Photoelastic analysis has great applicability in restorative dentistry as it allows prediction and minimization of biomechanical critical points through modifications in treatment planning. PMID- 19305248 TI - A new 3-phase therapy protocol in temporomandibular joint ankylosis: our experience. AB - Limitations in oral opening may be due either to intra-articular or extra articular ankylosis (pseudoankylosis). The principal means of therapy usually consist of 2 steps: surgical removal of the ankylotic block and immediate functional rehabilitation. In the postoperative period, however, immediate and adequate functional rehabilitation is not always possible because of pain and swelling of the temporomandibular joint, resulting in a very high risk of relapse. To prevent this, the authors introduce a third phase between the 2 already mentioned (surgery and functional therapy) in which the oral opening obtained with surgery is held in place while clinical conditions (pain and swelling) are allowed to stabilize before initiating intensive physiotherapy. Through the discussion of 18 clinical cases (8 of which were treated in the traditional way), the authors report on the intermediate treatment phase introduced by them between the surgical stage and the rehabilitation. Immediately after the operation, a handmade wedge, that is, a bite block of suitable size, is placed between the dental arches for a period of about 3 weeks, while appropriate pharmacological therapy was administered. The results showed an improvement of the mandibular movements both in opening and laterally, on the right and left sides, in patients treated with the new protocol. The proposed protocol holds the opening obtained with surgery until the clinical conditions make it possible to initiate intensive physiotherapy. In this way, the authors obtained better results in both recovery and in mandibular movements. PMID- 19305249 TI - The radix nasi island flap: a versatile musculocutaneous flap for defects of the eyelids, nose, and malar region. AB - A versatile musculocutan flap from the radix nasi region, the radix nasi island flap, is described. The flap has an axial blood supply derived from the dorsal nasal branch of the ophthalmic artery which is anastomosed to the terminal branch of the facial artery. The flap includes the skin, subcutaneous tissue, and procerus muscle. Ten patients, aged 50 to 86 years, have been reconstructed with this flap for defects in the nose (in 4 cases), midface (in 4 cases) and lower eyelids (in 2 cases). The mean flap size was 17 x 23 mm (range: 15 x 20 to 20 x 27 mm). All flaps fully survived. Additional complications and morbidity were not observed. The donor sites were closed a primarily closure in all cases. Follow-up ranged from 3 to 12 months (mean: 8.2 months). The radix nasi flap is a safe flap, has minimal donor site morbidity, and is especially suited for nasal and midface reconstruction in terms of attaining a suitable color and thickness. PMID- 19305250 TI - New velopharyngeal measurements at rest and during speech: implications and applications. AB - OBJECTIVES: To make proper management decisions, it is important to understand the cause of velopharyngeal inadequacy (VPI) in individual patients. Previous studies suggest that abnormal velar and pharyngeal structures as well as their ratios may contribute to VPI after primary palatal repair in some patients. The aim of the current study was to develop a new protocol to evaluate the velopharyngeal (VP) mechanism and the levator veli palatini muscles at rest and during speech production. A secondary goal was to investigate the correlations among VP measurements and craniofacial structures that may predict velopharyngeal motions during speech production. MATERIALS AND METHODS: Seventeen healthy adults were scanned with magnetic resonance imaging (MRI) techniques. Multiple MRI scans covered the craniofacial region, the VP mechanism, and the levator veli palatini muscles at rest and during sustained phonations of /a/, /i/, /z/, /m/, and /f/. Measurements of craniofacial and VP structures at rest and during speech productions were obtained using image analysis software. RESULTS: The maximal effective VP ratio, maximal pharyngeal constriction ratio, maximal velar stretch ratio, maximal velar height, and maximal levator shortening ratio were selected to represent the functional properties of the VP mechanism. The following predictors were found to partially explain the variances of corresponding measurements: rest effective VP ratio for the maximal effective VP ratio, maximal pharyngeal constriction for the maximal pharyngeal constriction ratio, and rest and maximal effective VP ratios for the maximal velar stretch. These predicting effects had implications in VP surgery and in understanding VP physiology. CONCLUSIONS: This study developed a practical imaging protocol to measure the VP mechanism at rest and during speech productions. New measurements such as the effective VP ratio, pharyngeal constriction ratio, velar stretch ratio, and levator shortening ratio may help to identify the cause of persistent VPI after palatal repair. Implications on the VP surgery were discussed. PMID- 19305251 TI - Orbital restoration surgery in the zygomaticotemporal and zygomaticofacial nerves and important anatomic landmarks. AB - A variety of etiologies may result in functional and aesthetic deficiencies requiring orbital reconstruction. Fractures of the zygomaticomaxillary complex in the acute stage are frequently accompanied by sensory disturbances of the zygomatic nerve (ZN). The purpose of the current study was to describe the anatomic and topographic landmarks of the ZN in 18 adult human cadavers regarding the localization and dimensions in the orbit. The zygomaticotemporal (ZTN) and zygomaticofacial nerves (ZFN) along the lateral wall of the orbit passed through the zygomaticotemporal and zygomaticofacial foramens, respectively. The angle between the ZTN and the ZFN within the orbit was approximately 42.21 degrees. The mean (SD) distance between the orbital opening of the ZTN and the meeting point of the ZTN was measured as 9.21 (5.18) mm. The mean (SD) distance between the orbital opening of the ZFN and the meeting point of the ZTN was calculated as 11.22 (4.25) mm. The mean (SD) distance between the orbital opening of the ZFN and the infraorbital margin of the orbit was 13.04. (3.21) mm. A detailed knowledge of the ZN's passage in the orbit is necessary for a surgeon while performing maxillofacial surgery. If these measurements are taken into account, there will be little surgical risk, and this will be helpful in identifying the extent of the operative field. PMID- 19305252 TI - Skull indices in a population collected from computed tomographic scans of patients with head trauma. AB - BACKGROUND: It is important to find out the skull's anatomic indices with computed tomography (CT), especially the skull's dimensions according to the patients age and sex, because it can give important results in neurosurgery. We will also describe the skull's parameters among the Kerman population. METHOD: Of the 2000 CT scans taken after head trauma injury in Kerman province from June 2002 to September 2006, we analyzed 1034 good quality films. The most frequent skull dimensions were measured on each film, and the data were computed and analyzed to assess the cranial indices in various ages and sexes. Two types of classifications were carried out: classic method, which uses cranial indices, and posterior fossa angle dimensions as an alternative method. Skull dimensions were statistically analyzed between groups, and any difference greater than 0.05 was considered significant. RESULTS: Of the 1034 films, 871 films belonged to the men and 184 to the women at age older than 5 years and younger than 70 years. Most of the CT films belonged to the patients with age older than 20 years and younger than 40 years in both sexes. Although the rates of various skull types were different according to the method of classification, the most frequent skull type was dolichocephalic followed by mesocephalic and brachycephalic in both sexes. In addition, in both sexes, the frequency of dolichocephaly was greater in children than adults, but brachycephaly was more frequent in adults than children. CONCLUSIONS: With CT scans as an accurate and suitable source of anthropometric evaluation of body dimensions, we found that, in our population, the most frequent skull type is dolichocephalic followed by mesocephalic, which seems to be closer to the Anglo-Saxon population considering the rates of dolichocephaly and mesocephaly. PMID- 19305253 TI - Clinical implications of the topography and distribution of the posterior superior alveolar artery. AB - The aim of this study was to clarify the arrangement of the anatomic courses and distribution of the intraosseous branch (IObr) of posterior superior alveolar artery. The anatomic variations in the topographic relationships were described to provide beneficial data to minimize injury to the IObr during surgical procedure of the buccal wall of the maxillary sinus. The IObrs in 42 hemifaces of embalmed Korean cadavers were examined. The courses of the IObr of the posterior superior alveolar artery were classified into 2 categories: the straight (type 1) and the U-shaped (type 2). The type 1 was the most common (78.1%), and the type 2 was observed in 21.9% of the specimens. The minimum mean height from the cervix to the IObr was 21.1 mm in the first molar region. The IObr ran at the lowest level from the maxillary sinus floor at the first premolar region. These anatomic findings in the current study could represent useful information for the various surgical procedures of the maxilla. PMID- 19305254 TI - Fibrolipoma of the tip of the nose. AB - Fibrolipoma is a rare benign tumor that usually presents as an asymptomatic, slowly growing mass with a firm or soft consistency clinically. Histopathologic findings show the proliferation of normal mature fat and fibrous tissue. We report a first case of the fibrolipoma of the nasal tip in a 24-year-old man. The lesion was totally excised and showed no evidence of recurrence after 6 months of follow-up. PMID- 19305255 TI - Carotid-cavernous fistula after functional endoscopic sinus surgery. AB - Carotid-cavernous fistulas (CCFs) are anomalous communications between the carotid arterial system and the venous cavernous sinus. They can arise because of spontaneous or trauma causes. Most caroticocavernous fistulas are of spontaneous origin and unknown etiology. Spontaneous CCF may also be associated with cavernous sinus pathology such as arteriosclerotic changes of the arterial wall, fibromuscular dysplasia, or Ehler-Danlos syndrome. Traumatic CCFs may occur after either blunt or penetrating head trauma. Their clinical presentation is related to their size and to the type of venous drainage, which can lead to a variety of symptoms, such as visual loss, proptosis, bruit, chemosis, cranial nerve impairment, intracranial hemorrhage (rare), and so on. Treatment by endovascular transarterial embolization with electrolytically detachable coils is a very effective method for CCF with good outcomes. Carotid-cavernous fistulas have been rarely reported after craniofacial surgery and are uncommon pathologies in otolaryngology practice. In this study, we report a 40-year-old woman with CCF secondary to blunt trauma of functional endoscopic sinus surgery. PMID- 19305256 TI - Aspiration of two permanent teeth during maxillofacial injuries. AB - Aspiration of a tooth in maxillofacial injuries is a known complication necessitating prompt recognition and early treatment to minimize potentially serious and, sometimes, even fatal consequences. Here, we report a rare and unusual case in its presentation, the patient having aspirated 2 permanent teeth after maxillofacial, cervical vertebrae, and thoracic region crush injuries due to a motor vehicle accident. The diagnosis of teeth aspiration was delayed until 2 weeks after the event. An aspirated anterior tooth was expectorated by the patient himself, and the other aspirated anterior tooth was removed by flexible bronchoscopy. The paper also emphasizes that clinicians must be aware of dental injury resulting from maxillofacial injuries and account for all teeth as part of their evaluation, keeping an open mind as to where a missing tooth might be located. A rapid diagnosis depends on high clinical suspicion, clinical signs, and radiologic findings. PMID- 19305257 TI - Methods used for assessing stresses in buccomaxillary prostheses: photoelasticity, finite element technique, and extensometry. AB - The authors describe a literature revision on assessing stresses in buccomaxillary prostheses photoelasticity, finite element technique, and extensometry. They describe the techniques and the importance for use of each method in buccomaxillary prostheses with implants and the need of accomplishing more studies in this scarce literary area. PMID- 19305258 TI - Correction of depressed forehead with BoneSource in cleidocranial dysplasia. AB - Cleidocranial dysplasia is a well-documented rare congenital disorder of the bone characterized by abnormalities of the skull, clavicle, and dentition. Despite numerous observations, there are still comparatively few reports regarding patients with cleidocranial dysplasia that focus on the impact of reconstruction of these defects on facial aesthetics. We report a 19-year-old woman with opened metopic and sagittal sutures and delayed closure of the anterior fontanelle. Through bicoronal incision and pericranial elevation, the defect was fully exposed. BoneSource was used to fill the cranial defect and was contoured to the desired cosmetic outcome. During a 24-month follow-up period, no absorption or recurrence was seen, and the patient was satisfied. PMID- 19305259 TI - A rare complication of nasotracheal intubation: accidental middle turbinectomy. AB - In this paper, we are presenting a rare case of accidental middle turbinectomy, a complication of nasotracheal intubation. We have reviewed the literature and addressed important parameters on nasotracheal intubation to avoid damage to the turbinates and its possible serious complications. PMID- 19305260 TI - Craniofacial gunshot injury resulting in pseudoaneurysm of the left internal maxillary artery and Collet-Sicard syndrome. AB - Facial gunshot is a challenging medico-surgical emergency because of the complex neurovascular structure in the area. We present a patient who sustained a facial gunshot injury, with massive nasal and oral bleeding that resulted in shock. We review the management of gunshot injury to the internal maxillary artery, both in acute and chronic stages. In addition, we describe the clinical course of the rarely occurring Collet-Sicard syndrome, which involves injury to the 9th, 10th, 11th, and 12th cranial nerves associated with gunshot injury. PMID- 19305261 TI - Otalgia due to the pleomorphic adenoma of uvula. AB - We report an unusual presentation of a salivary pleomorphic adenoma in the uvula. The adenoma caused otalgia and hypernasal speech in a patient without additional symptoms. The tumor was surgically excised with safe margins of the healthy tissue. After the excision of the mass, the new uvula was reconstructed through modified radiofrequency-assisted uvulopalatoplasty technique. After the treatment, the patient has remained disease-free for 1 year. PMID- 19305262 TI - Reconstruction of a congenital nasal deformity using skin tags as a chondrocutaneous composite graft. AB - Nasal reconstruction has key functional aims of achieving an optimal esthetic result while maintaining a patent airway. For the distal third of the nose, these goals are achieved by replacement of lining, osseocartilaginous support, and soft tissue coverage. The use of skin tags for reconstruction of the ear have been well documented, yet to our knowledge, there are no reported cases of their use in reconstruction of the nose. Skin tags, also referred to as acrochordon, fibroepithelial polyps, and pedunculated fibromas are benign soft, fleshy, sessile, or pedunculated lesions ranging from 1 to 5 mm in diameter. We report a novel approach to distal nasal reconstruction that arose opportunistically as a result of coincident pathologies. PMID- 19305263 TI - Third molar extraction with massive hemorrhage treated by embolization. AB - Impaction of the third molar is relatively frequent in oral and maxillofacial surgery, and its removal is a usual operation with mostly unremarkable outcome. We report a case of bleeding that occurred after a left upper third molar extraction, which necessitated in emergency an angiography with embolization. PMID- 19305267 TI - Reconstruction of cranial bone defects using a quick-setting hydroxyapatite cement and absorbable plates. PMID- 19305268 TI - Variations in autologous blood transfusion systems. PMID- 19305269 TI - Transtrochanteric anterior rotational osteotomy for osteonecrosis of the femoral head in patients 20 years or younger. AB - BACKGROUND: For young patients with osteonecrosis of the femoral head, especially those younger than 20 years, a joint-preserving operation may be one of the surgical treatment options to be considered. We investigated the clinical and radiological results of transtrochanteric anterior rotational osteotomy for the treatment of osteonecrosis of the femoral head in patients 20 years or younger. METHODS: Between 1976 and 2001, a transtrochanteric anterior rotational osteotomy was performed in 28 hips of 23 patients aged 20 years or younger with osteonecrosis of the femoral head. Among them, 27 hips of 22 patients with a minimum follow-up duration of 2 years were investigated (follow-up rate, 96.4%). They included 7 males and 15 females with a mean age of 16 years at the time of surgery. The mean follow-up period was 14.7 years (range, 2.0-31.7 years). The clinical assessment was made based on the Merle d'Aubigne-Postel score. The postoperative intact ratio, joint-space narrowing, and progression of collapse were all investigated radiographically. RESULTS: Two (7.4%) of the 27 hips required total hip arthroplasty because of restrictions in the range of motion, whereas the original hip joints in 25 (92.6%) of the 27 hips were preserved at the time of final follow-up. The mean preoperative Merle d'Aubigne-Postel score was 10.4 points, which improved to 15.9 points at the final follow-up (P < 0.0001). The mean postoperative intact ratio was 49.8% (range, 30-100). Five (18.5%) of the 27 hips had a progressive joint-space narrowing, but progression of the collapse was not observed in anyhips. CONCLUSIONS: Transtrochanteric anterior rotational osteotomy appears to be a useful joint-preserving operation for patients younger than 20 years. LEVEL OF EVIDENCE: Level 4 (Therapeutic Study). PMID- 19305270 TI - Difficult-to-treat Ortolani-positive hip: improved success with new treatment protocol. AB - BACKGROUND: The aim of this paper was to review the outcomes of treatment of dislocated but reducible (Ortolani-positive) hips. METHODS: A comparative retrospective review was conducted of all patients aged younger than 6 months who presented for treatment of developmental dysplasia of the hip for two 10-year periods. The inclusion criteria consisted of a clinical evaluation documenting Ortolani-positive examination, no previous treatment, and an initial treatment with Pavlik harness only. Patients with insufficient follow-up, neuromuscular disease, provocative dislocatable hips, or dislocated but not reducible hips were excluded. In group 1 (1984-1994), treatment involved initiation of Pavlik harness and parental education at the time of initial diagnosis. Group 2 (1997-2007) was treated the same as group 1 with the addition of serial orthopaedic office-based ultrasound examinations and use of a hip abduction orthosis in hips remaining unstable after 3 weeks in the Pavlik harness. Records were assessed for successful achievement and maintenance of hip reduction, need for and type of further treatment, and development of avascular necrosis. RESULTS: In group 1, the mean time of follow-up was 28 months. Forty-four (85%) of 52 hips were treated successfully. Eight patients required further operative treatment. In group 2, the mean follow-up to the final evaluation was 34 months. Forty-one (93%) of 44 hips were treated successfully. Three hips required further treatment. There were no documented cases of avascular necrosis in patients treated for orthoses only in either group. CONCLUSIONS: In 1994, we presented a series of 52 Ortolani-positive hips (group 1) with a success rate of reduction of 85%. Now, our protocol for treating Ortolani-positive hips has evolved to include serial orthopaedic office-based ultrasound in all patients and use of a hip abduction orthosis in hips remaining unstable after 3 weeks in a Pavlik harness. Three previous studies at major centers reported successful reduction in only 63% to 71%. In group 2, our current success rate of 93% exceeds that previously reported by us and is significantly greater than these 3 previous studies (all P < 0.006). Furthermore, our current treatment module has decreased our rate of open reduction from 10% in group 1 to 5% in group 2. LEVEL OF RELEVANCE: Therapeutic study, level 3. PMID- 19305271 TI - Hospitalized cases of child abuse in America: who, what, when, and where. AB - BACKGROUND: Orthopaedic surgeons need to recognize features of child abuse. The purpose of this study was to identify common characteristics of child abuse and to delineate features of physical injury cases that would raise concern for child abuse. METHODS: A nationally representative sample of all pediatric inpatient discharges for calendar years 1997, 2000, and 2003 was queried for physical injury diagnoses. Cases were identified as those with a diagnosis of child abuse and controls were those without the diagnosis of child abuse. Incidence of coded child abuse and a comparison of rates across time, demographics, and injury pattern were determined. Cases were compared with controls, and relative risks for coded child abuse were computed using the generalized linear model specifying the Poisson distribution and a log link. RESULTS: There were 665,195 physical injury cases identified. Of these, 11,554 (1.74%) had a diagnosis of child abuse. Fracture requiring orthopaedic management was present in 28% of the child abuse cases. The 3 strongest demographic predictors of coded child abuse were age younger than 1 year, age 1 to younger than 2 years, and Medicaid as primary payer with adjusted relative risks of 11.46, 3.07, and 1.99, respectively. Winter and weekday presentation were significantly higher for coded child abuse. Fractures with the greatest adjusted relative risks for coded child abuse were rib or sternum (5.34) and scapula (3.22). Pelvic fracture was the only fracture significant for lowered adjusted relative risk of coded child abuse. CONCLUSIONS: This study shows that young age continues to be a strong predictor of child abuse in the setting of physical injury. Supporting features include Medicaid as primary payer and winter or weekday presentation. Orthopaedists should be particularly aware of child abuse as 28% of cases had a fracture requiring orthopaedic management. LEVEL OF EVIDENCE: Prognostic Study, Level III (case control study). PMID- 19305272 TI - Proximal humeral fractures in children and adolescents. AB - BACKGROUND: The purpose of the study was to investigate possible reasons for the failure of closed reduction of proximal humerus fractures in children and adolescents. We assessed the rate of soft tissue entrapment, and we also investigated the long-term clinical and radiological results after an age- and deformity-focused treatment regimen according to national guidelines. METHODS: Forty-three patients were included in the study. Ten (mean age, 12.4 years; range, 6-16 years) of the patients were treated conservatively. The remaining 33 patients (mean age, 14 years; range, 6-18 years) were treated surgically (n = 2 Neer grade 2, n = 16 Neer grade 3, and n = 15 Neer grade 4) with either closed (n = 16) or open reduction with internal fixation. In 17 fractures, closed anatomical reduction of the fracture under general anesthesia was not possible. Subsequent open reduction and Kirschner wire or screw fixation (n = 12) or plate fixation (n = 5) was necessary. In 9 of these 17 fractures (5 fractures were totally displaced fractures), closed reduction was impossible because of the entrapment of periost (n = 2) or the biceps tendon with parts of the periost (n = 7). At follow-up, the clinical assessment included a structured interview, a detailed physical examination, and the assessment of overall shoulder function with the Constant score. RESULTS: Operative and postoperative complications did not occur. All surgically treated fractures anatomically reduced and healed without loss of reduction. At a mean follow-up of 39 months (range, 12-118 months), all patients who were evaluated had excellent results according to the Constant score and had a normal range of motion and excellent strength of the shoulder joint. CONCLUSIONS: A failed closed reduction should be interpreted as a possible soft tissue entrapment most likely because of the long biceps tendon. Those cases should be addressed with open reduction and removal of the entrapped structures. If anatomical reduction is achieved and maintained until fracture healing, excellent functional and radiological results can be expected from an age- and deformity-focused treatment regimen for children and adolescents with proximal humeral fractures. LEVEL OF EVIDENCE: Level 4 (Therapeutic study). PMID- 19305273 TI - A new closed reduction technique for the treatment of radial neck fractures in children. AB - Displaced radial neck fractures in the skeletally immature patient are uncommon and can be difficult to reduce. We describe a new technique for the closed reduction of radial neck fractures in children. One of the authors (R.M.) has used this technique in treating 6 children with displaced radial neck fractures. This was a retrospective review carried out at a tertiary care hospital in Winnipeg, Manitoba, Canada. All charts and radiographs were reviewed. One patient underwent an ultrasound assessment to assist reduction (case 4). Average age of the patients was 9 years and 6 months. Average follow-up was 4.3 months. Closed reduction was successful in all patients. No patient went on to an open reduction. The results in this series using a newly described technique are encouraging, and there is consideration for its continued use. PMID- 19305274 TI - Head-shaft angle measurement in children with cerebral palsy. AB - BACKGROUND: It has been observed in children with cerebral palsy (CP) that the femoral head rests in a valgus position as compared to the proximal neck and/or shaft. The purpose of this study was to compare the head-shaft angle (HSA) as a measure of femoral head valgus in children with CP (group 1) with a subset of children with CP who have more significant hip subluxation (group 2, as demonstrated by the need for surgery to correct the deformity), with age-matched (group 3) and historical controls. METHODS: There were a total of 39 patients (70 hips), 15 patients in group 1, 10 in group 2, and 14 in group 3. The HSA was measured as described by Southwick (Southwick WO. Osteotomy of the lesser trochanter for slipped capital femoral epiphysis. J Bone Joint Surg Am. 1967;49A:803-835). Interrater and intrarater reliabilities and the variation of these measures with rotation were assessed. RESULTS: The mean HSA in group 1 was 160.8 degrees; group 2, 170.3 degrees; group 3, 152.5 degrees; and historical control, 146.7 degrees. The difference was statistically significant between groups 1 and 2 and between both CP groups and the control groups. Intrarater and interrater reliabilities were found to be low, and variation with rotation was found to be minimal. CONCLUSIONS: This study demonstrates that HSA is greater in children with CP than in typically developing children and that this is more pronounced in children with CP who are at risk for eventual subluxation. Evaluation of the HSA may be prudent in children with CP. PMID- 19305275 TI - Iliopsoas tenotomy at the lesser trochanter versus at the pelvic brim in ambulatory children with cerebral palsy. AB - BACKGROUND: Progressive hip flexion deformity is a common problem in ambulatory children with spastic cerebral palsy, causing static and dynamic deformity. The iliopsoas muscle is recognized as a major deforming force in the development of this problem. Many clinicians address this problem by lengthening the iliopsoas, either in an intramuscular location at the pelvic brim or by complete tenotomy at the lesser trochanter. The goal of this study was to compare the outcomes of patients with ambulatory cerebral palsy who had intramuscular lengthening at the pelvic brim to those who underwent complete release of the iliopsoas tendon at the level of the lesser trochanter. METHODS: Twenty patients were included in the study, 11 of whom had iliopsoas release at the lesser trochanter (group 1) and 9 of whom had intramuscular lengthening at the pelvic brim (group 2). All patients had physical examinations, plus kinematic and kinetic analyses in our gait laboratory before and 1 year after surgery. RESULTS: Hip flexion contracture was decreased significantly only in group 1, although there was a trend of decrease in group 2. There was a significant increase in maximum hip extension in terminal stance and a reciprocal decrease in maximum swing phase hip flexion in group 1, with a similar trend that did not reach significance in group 2. Stride length increased significantly in both groups. There was no significant change in power generation of hip flexion during the swing phase in either group. CONCLUSIONS: We found improved static and dynamic parameters of hip extension after iliopsoas lengthening and did not detect any adverse kinematic or kinetic change in hip function after surgery. The improvement was more robust in the group who underwent release at the lesser trochanter. Because there are no adverse effects of iliopsoas release from the lesser trochanter and the improvement in hip extension is greater, this approach should be considered in ambulatory patients with spastic diplegia when a hip flexor weakening procedure is considered. LEVEL OF EVIDENCE: Comparative cohort study, level III, case-control study. PMID- 19305276 TI - Prevalence and risk factors in postoperative pancreatitis after spine fusion in patients with cerebral palsy. AB - BACKGROUND: Postoperative pancreatitis is a recognized complication after spine fusion surgery in scoliosis management. There are established risk factors for the development of postoperative pancreatitis for patients with scoliosis in general, but no such factors have been established in patients with cerebral palsy (CP) who have scoliosis. The aim of this study was to assess the association between preoperative comorbidities and other parameters in the development of pancreatitis after spinal fusion surgery in a sample of children with CP. METHODS: A retrospective cohort design was used to assess the association between postoperative pancreatitis and preoperative comorbidities and other covariates that might predispose to pancreatitis. The study participants included 355 patients, with a mean age of 13.8 years (range, 5.6-21 years) with CP, who received spine fusion with rod instrumentation. We used the chi statistic, mean, and SD to describe study variables that were categorical and continuous, respectively. To test the null hypothesis of no association between our outcome variable (postoperative pancreatitis) and comorbidities and other covariates as predictors, we used a binomial regression model. To simultaneously adjust for confounding effects in the relationship between our outcome and the independent or predictor variables, we used a multivariable binomial regression model. RESULTS: Of the 355 patients who underwent spine fusion, 109 developed postoperative pancreatitis (prevalence, 30.1%). Patients who developed pancreatitis had prolonged number of fasting days (until oral or G-tube feeding initiated; 7.9 vs 5.2 days) and longer hospital stay (23.1 vs 15.6 days). In the univariable binomial regression model, patients with CP who had gastroesophageal reflux disease (GERD) and feeding difficulty were more likely to develop pancreatitis than those without this condition (risk ratio [RR], 1.57; 95% confidence interval [CI], 1.10-2.28, respectively). Likewise, in this model, gastrointestinal tube and reactive airway disease (RAD) were statistically significantly associated with postoperative pancreatitis. Patients with CP who had gastrointestinal tube were 61% more likely to develop postoperative pancreatitis, whereas those with RAD were 54% (RR, 1.61; 95% CI, 1.01-2.55 and RR, 1.54; 95% CI, 1.13-2.10, respectively). However, there was a clinically relevant but nonstatistically significant association between seizure and postoperative pancreatitis (RR, 1.72; 95% CI, 0.96-3.06). After adjustment for the confounding variables in the multivariable model, GERD with feeding difficulties persisted as a single most significant and potent predictor of postoperative pancreatitis (adjusted RR, 1.52; 95% CI, 1.01-2.29). Consequently, patients with CP who had GERD were 52% more likely to develop postoperative pancreatitis. Likewise, there was a statistically significant 49% increase in the risk of postoperative pancreatitis in patients with CP who had RAD (adjusted RR, 1.49; 95% CI, 1.10-2.04). CONCLUSIONS: Pancreatitis is a major cause of morbidity after spinal fusion surgery in patients with CP. Patients with preoperative GERD with feeding difficulties and RAD had a higher risk of developing postoperative pancreatitis. Postoperative pancreatitis causes delays in feeding and increases the duration of hospitalization. Clinicians should be aware of the roles of GERD and RAD, as well as seizure and/or antiepileptics in the development of postoperative pancreatitis in patients with CP undergoing spinal fusion. LEVEL OF EVIDENCE: Level III. PMID- 19305277 TI - Acute pediatric upper extremity compartment syndrome in the absence of fracture. AB - BACKGROUND: To determine the etiologies and outcomes associated with acute pediatric upper extremity compartment syndrome in the absence of fracture. METHODS: A retrospective review was performed looking at children treated for acute upper extremity compartment syndrome in the absence of fracture at a major teaching hospital. Reason for admission, age, etiology, sensorium, time to fasciotomy, involved compartments, secondary procedures, and functional outcome were recorded. RESULTS: A total of 14 extremities in 13 children with acute compartment syndrome in the absence of fracture were identified over a 22-year period at this single institution. There were 8 boys and 5 girls, with an average age of 7.2 years. Average follow-up was 22 months. Ten patients were being managed in the intensive care unit and had an obtunded sensorium. The cause was iatrogenic in 8 patients, and 2 of these resulted in loss of the involved limb. Six patients required 9 secondary procedures, including 4 amputations, 3 contracture releases, and 2 skin grafts. Of the 3 patients who required a total of 4 amputations, 2 of the patients were in the intensive care unit, and all were younger than 3 years. Only 7 patients had normal hand function. Upon comparing patients with a normal outcome versus those with an abnormal outcome, there was a statistically significant difference if surgery was performed in shorter than 6 hours (P = 0.033). CONCLUSIONS: This problem is often iatrogenic in etiology and can be diagnosed late in this population. An increased level of vigilance should be adopted for this entity because the final outcome can be catastrophic for both the patient and the hospital and early fasciotomy is associated with improved results. LEVEL OF EVIDENCE: Level IV Case Series. PMID- 19305278 TI - Congenital elevation of the scapula: surgical treatment with Mears technique. AB - BACKGROUND: Several procedures have been described for the treatment of congenital elevation of the scapula. The method proposed by Mears in 2001 seems to provide highly favorable functional and cosmetic results. To date, there are no studies in literature that supported the initial results obtained by this author. The aim of our study is to evaluate the appearance and range of motion obtained with Mears technique. METHODS: We evaluated prospectively 21 consecutive patients treated from June 2001 to June 2006. Fourteen of them (5 males and 9 females) with Cavendish grade III and IV were treated with said technique. Mean age was 6.7 years (range, 4-10 years). Mean follow-up was 45 months (range, 12-74 months). The operative results were evaluated on cosmetic and functional criteria. Functional evaluation includes preoperative and postoperative range of motion and strength. Appearance evaluation was performed according to Cavendish Scale. RESULTS: Range of motion improved significantly. On average, flexion improved from 83.9 degrees (range, 50-120 degrees) to 152.1 degrees (range, 110 180 degrees) (P < 0.001), and abduction improved from 81 degrees (range, 50-120 degrees) to 145 degrees (range, 100-180 degrees) (P < 0.001). Appearance improved by a mean of 2 levels on the Cavendish Scale. In 2 patients, a second operation was performed to remove a residual exostosis. Two patients developed keloid scars. All patients expressed satisfaction with operative results. CONCLUSIONS: Mears technique provides significant improvement in range of motion and a considerable appearance improvement with a low morbidity. Further clinical experience will be needed to confirm these outcomes. PMID- 19305279 TI - Minimizing tourniquet pressure in pediatric anterior cruciate ligament reconstructive surgery: a blinded, prospective randomized controlled trial. AB - BACKGROUND: Tourniquet cuff pressures in pediatric patients are commonly set at standard pressures. Recent evidence on adult subjects has shown that safer and more effective cuff pressures can be achieved by measuring limb occlusion pressure (LOP) and using a wide contour cuff. There is little evidence validating these techniques in children. The primary objective of this study was to evaluate if a difference in tourniquet cuff pressure can be achieved in a pediatric population using a wide contour cuff in conjunction with measured LOP when compared with a standard cuff and pressure. METHODS: Subjects aged 10 to 17 years that underwent anterior cruciate ligament repair were included and randomized into either the control group or the experimental LOP group using variable block randomization. The tourniquet cuff was inflated to 300 mm Hg in the control group or to the recommended tourniquet pressure based on LOP measurement in the LOP group. The surgeon was blinded to cuff selection, application, and pressure throughout the surgical procedure. Immediately after the surgical procedure, the surgeon rated the quality of the bloodless field on a visual analog scale. This study was powered as an effectiveness trial, and intention to treat analysis was used. RESULTS: After a planned interim analysis at midpoint, complete data were recorded for 11 (control group) and 10 (LOP group) patients. The quality of the surgical field was not different between the groups (P = 0.053). There was a statistically significant difference in the mean cuff pressure between the control (300 mm Hg) and the LOP (151 mm Hg) groups (P < 0.001). We ran the same analysis comparing the LOP data with the hypothetical control data of 250 mm Hg, and our results remained statistically significant (P < 0.001). CONCLUSIONS: The use of an automatic LOP measurement with the use of wide contour cuffs can significantly reduce mean tourniquet cuff pressures in pediatric patients compared with the typical practice of 300 or 250 mm Hg without compromising the quality of the surgical field. LEVEL OF EVIDENCE: Level 1, prospective randomized controlled trial. PMID- 19305280 TI - Interobserver and intraobserver reliability in the evaluation of mechanical axis deviation. AB - BACKGROUND: The lateral distal femoral angle (LDFA), the medial proximal tibial angle (MPTA), and the mechanical axis deviation (MAD) are commonly used in the evaluation of lower extremity deformities. The interobserver and intraobserver reliabilities of these measurements have not been evaluated. METHODS: Three groups of observers with different levels of experience (5 attending pediatric orthopaedic surgeons, 5 orthopaedic chief residents, and 5 interns) measured the LDFA, MPTA, and MAD on 35 full-length standing anteroposterior teleroentgenograms (56extremities). Relatively equal numbers of extremities with varus, valgus, and normal alignments were chosen to represent a spectrum of lower extremities that might be encountered clinically. Measurements were performed in random order by each observer on 2 separate occasions separated by at least 2 weeks. RESULTS: The interobserver and intraobserver reliabilities for each of the measurements regardless of the level of experience was greater than or equal to 0.90. The interobserver differences were slightly greater than the intraobserver differences for all measurements. The overall mean interobserver differences for angular and MAD measurements were within 1.6 degrees and 3.1 mm, respectively. The overall mean intraobserver differences for angular and MAD measurements were within 1.4 degrees and 1.9 mm, respectively. Reliability and mean differences were consistent regardless of the level of experience. CONCLUSIONS: Measurement of the LDFA, MPTA, and MAD demonstrated excellent intraobserver and interobserver reliabilities regardless of the experience of the observer. PMID- 19305281 TI - Hemiepiphyseodesis for late-onset tibia vara. AB - BACKGROUND: The amount of correction obtained with tibial and/or femoral hemiepiphyseodesis in late-onset tibia vara is quite variable. The purpose of this study is to identify preoperative factors which might help to predict the amount of correction that is obtainable. METHODS: Fifty-three patients (67 knees) have been treated with hemiepiphyseodesis and followed until skeletal maturity or a secondary realignment procedure before maturity. The amount of correction obtained was correlated with multiple preoperative factors including age, body weight, and degree of deformity. RESULTS: In 65 limbs treated with tibial hemiepiphyseodesis, the average correction in the tibia was 9 degrees, ranging widely from 33 degrees of correction to 6 degrees worsening of the deformity. In 22 limbs treated with femoral hemiepiphyseodesis, the average correction in the femur was 8 degrees (range, 0-19 degrees). Ultimately, 19 of the 67 limbs have been treated with an osteotomy and 10 more were left with greater than 10 degrees of varus deformity at maturity. In addition, 14 of the 67 limbs overcorrected into excessive valgus and/or required medial hemiepiphyseodesis procedures to prevent overcorrection. Using multiple regression analysis it was found that younger age and lesser deformity were weakly predictive of greater correction with tibial hemiepiphyseodesis. No factors were statistically significant predictors of femoral correction. CONCLUSIONS: Variable amounts of correction are obtained with hemiepiphyseodesis in patients with late-onset tibia vara. Even very large amounts of correction may be obtainable in some cases, but unfortunately preoperative factors such as age, weight, and degree of deformity cannot be relied upon in clinical practice to predict outcome. Surgical decision making must weigh the safety and simplicity of this procedure against the much more extensive but much more predicable realignment obtained with osteotomy procedures. LEVEL OF EVIDENCE: Level IV; retrospective and prospective longitudinal evaluation of a case series. PMID- 19305282 TI - Fetoscopic release of extremity amniotic bands with risk of amputation. AB - With the advent of less-invasive fetal surgery techniques, nonlethal disorders are considered amenable to intrauterine treatment. Extremity amniotic band syndrome fulfils the criteria of intrauterine disorders conformable with fetal treatment: capability of prenatal diagnosis and severity. We report 2 cases of extremity amniotic bands with risk of limb amputation released fetoscopically with YAG laser. PMID- 19305283 TI - Osteotomies of the foot for cavus deformities in children. AB - BACKGROUND: The cavovarus foot has been defined as plantar flexion of the first ray. The usual cause is due to a muscle imbalance. The purpose of this study was to report our experience with selective, joint-sparing osteotomies of the foot that address each deformity in the cavovarus foot in a stepwise fashion. Most bony procedures for correction of cavus feet have centered on osteotomies across multiple joints or fusions. METHODS: We report on stepwise osteotomies: (1) closing wedge to the first metatarsal, (2) opening plantar wedge of the medial cuneiform, (3) cuboid closing wedge, (4) and as needed second and third metatarsal osteotomies, calcaneal sliding osteotomies, and plantar fasciotomy and peroneus longus-to-brevis transfer. We measured all feet radiographically and clinically. RESULTS: We studied 20 feet in 13 patients with multiple etiologies. Nearly all feet were graded good to excellent on our outcome scale. Correction in Meary and Hibb angles was observed. There were no significant complications. CONCLUSIONS: By performing a double osteotomy on the first ray (cuneiform and metatarsal), the cavus can be nearly fully corrected. The cuboid osteotomy provides increased mobility of the forefoot. The sliding calcaneal osteotomy should be used to improve any residual hindfoot varus. We recommend transferring the peroneus longus to brevis to balance the paralytic foot. The cavus foot needs to be addressed at the apex, while sparing the midtarsal joints and avoiding fusion. This sequence of osteotomies addresses all of the components of a cavus foot. LEVEL OF EVIDENCE: Therapeutic study-level IV. PMID- 19305284 TI - Perioperative blood transfusion requirements in pediatric scoliosis surgery: the efficacy of tranexamic acid. AB - BACKGROUND: Previous studies have noted that the use of antifibrinolytic medications can help reduce blood loss and transfusion requirements during cardiac, total joint arthroplasty, and spine surgery. Tranexamic acid (TXA) has been investigated in these patient groups but consensus with respect to the dosing regimen has not been achieved, especially in the pediatric scoliosis literature. The purpose of this study was to compare the effects of 2 TXA dosing regimens on reducing transfusion requirements. METHODS: A retrospective chart review was performed on all idiopathic scoliosis patients undergoing posterior spinal instrumentation and fusion from 2005 to 2006 to determine total perioperative transfusion requirements. Transfusion requirements for those patients receiving either a low (10 mg/kg loading, 1 mg/kg/h infusion) or high (20 mg/kg loading, 10 mg/kg/h infusion) dose of TXA were compared. RESULTS: High dose TXA (n = 11) showed a trend toward a reduction in transfusion requirements compared with the low dose (n = 15) for idiopathic scoliosis patients undergoing posterior only instrumentation and fusion (687.9 +/- 778.1 mL vs 1372.6 +/- 1077.3 mL; P = 0.07; 95% confidence interval for the mean difference, -66.3 mL to 1435.7 mL). Although substantial, this difference was underpowered to show a difference. CONCLUSIONS: The use of the higher dose of TXA resulted in a 50% reduction in transfusion requirements for idiopathic scoliosis patients. Given previous studies, there appears to be a dose-response effect. A prospective dose ranging study is now required to determine the optimal dose for pediatric patients with idiopathic scoliosis. LEVEL OF EVIDENCE: III, retrospective cohort study. PMID- 19305285 TI - Effectiveness of free gracilis muscle flaps in the treatment of chronic osteomyelitis with purulent fistulas at the distal third of the tibia in children. AB - BACKGROUND: Chronic purulent infection at the distal third of the tibia resulting from of a previous hematogenous osteomyelitis in children is a severe infection which is seldom reported in the literature and remains a difficult challenge for orthopaedic surgeons. METHODS: A 6-year retrospective review from September 2001 to October 2006 (institutional review board-approved) was performed to evaluate postoperative surgical results of this type of infection using a free gracilis muscle flap. RESULTS: A total of 6 children with an average age of 9.8 years were included. The infective period of purulent fistulas ranged from 6 to 31 months (mean 17.7). Staphylococcus aureus were reported in all wounds (6/6) and were commonly combined with Escherichia coli (5/6). All of the free gracilis muscle transfers were successful (6/6 flaps). No postoperative recurrence of infection was observed with an average postoperative follow-up period of 3 years. All patients were also able to satisfactorily demonstrate walking and standing functions. CONCLUSIONS: Both the radical debridement of infected tissues and restoration of ample blood supply using free muscle flaps are the keys to success. Based on its favorable form, the free gracilis muscle flap can be considered a suitable muscle for use in reconstruction. LEVEL OF EVIDENCE: IV. PMID- 19305286 TI - Deep venous thrombosis in a child: an unusual presentation of an osteochondroma. AB - Deep vein thrombosis is very uncommon in the pediatric population. We describe an unusual case of deep vein thrombosis caused by a femoral osteochondroma in a 13 year-old adolescent girl with known multiple hereditary exostoses. We discuss the optimum investigations to determine the site of venous obstruction, which is crucial when there are several osteochondromata in the affected limb, which could be implicated in causing the obstruction to venous flow. PMID- 19305287 TI - Ragab AA, Stewart SL, Cooperman DR. Implications of Subtalar Joint Anatomic Variation in Calcaneal Lengthening Osteotomy. J Pediatr Orthop. 2003;23:79-83. PMID- 19305288 TI - Re: Orthopaedic complications of myotubular myopathy. PMID- 19305289 TI - Center of the femoral head in children. PMID- 19305291 TI - Who do you see when you reflect? PMID- 19305293 TI - Landstuhl Regional Medical Center: traumatic brain injury screening program. PMID- 19305294 TI - When West meets East: the experience of the Society of Trauma Nurses' Nursing delegation's visit to China with the People to People Citizen Ambassador Program. AB - In October 2008, 39 nurses traveled to China as part of the People to People Citizen Ambassador Program Emergency and Trauma Nursing delegation. The group compared and contrasted emergency and trauma nursing practices in the United States with those in China. Similarities include the methods of nursing education, the focus on patient and family education, and the passion of the nurses for delivering optimal care to their patients. Differences include the Chinese nurse's use of traditional Chinese medicine, the lack of modern technology, and low wages of healthcare personnel. The trip emphasized to the delegation the imperative for all nurses, regardless of their location in the world to reach out to one another to share best practices and their love of nursing. PMID- 19305295 TI - Changing physiological status predicts severe injury and need for specialized trauma center resources. AB - BACKGROUND: This study evaluated the association between changing physiological status (delta data) with severe injury (SI) or need for trauma center resources (TCR). METHODS: Prehospital and emergency department arrival weighted RTS (RTSw) were computed for patients with complete records entered into the registry from 2002 to 2004 (n = 23,753). Physiological change was classified as unchanged, deteriorated, or improved (PreRTSw vs EDRTSw). Performance of delta data was evaluated using standard epidemiological approaches and multiple logistic regression. FINDINGS: Deterioration status predicted SI (operating room [OR] = 1.38) and TCR (OR = 2.09). Improved status predicted TCR (OR = 1.27). CONCLUSIONS: Delta data independently predicted both SI and TCR. PMID- 19305296 TI - Traumatic bladder laceration in the absence of pelvic fracture. AB - Traumatic bladder laceration commonly occurs because of blunt trauma with associated pelvic fracture. The incidence of bladder laceration without pelvic fracture is less than 10%. Although high mortality (44%) existed in the past, a timely diagnosis with appropriate medical and surgical management now offers an excellent outcome. Early clinical suspicion, appropriate and reliable radiologic studies, and prompt surgical intervention, when indicated, are the keys to successful diagnosis and management. PMID- 19305297 TI - Care of the trauma patient beyond the emergency department: a patient care standard to guide bedside nurses. AB - Injuries sustained from illicit drug use or alcohol intoxication are common in emergency departments. Ongoing assessments of psychosocial issues in trauma patients are imperative, even after the patient leaves the specialized area of the emergency department. Oftentimes, bedside nurses are ill prepared to identify the subtle clues of deeper psychosocial issues in complex patients such as trauma patients. This article focuses on the rationale for the development of a patient care standard to guide the bedside staff nurse in the care of the trauma patient. An example of a multiple trauma diagnosis-related patient care standard is presented. PMID- 19305298 TI - Midlevel practitioner role evolution in an American College of Surgeons-verified trauma surgery service: the 23-year experience at Hurley Medical Center. AB - The purpose of this study was to evaluate our 23-year experience with midlevel practitioners in an American College of Surgeons-verified trauma surgery service in the absence of a surgical residency. A retrospective review of the utilization of trauma surgery midlevel practitioners was performed at a community hospital American College of Surgeons-verified trauma center from 1985 to 2007. The trauma surgery service has expanded in volume, acuity, and services offered over the 23 year period. This model has allowed us to establish effective and efficient surgical coverage while growing the trauma program, as well as general surgery and bariatric services. PMID- 19305299 TI - Temporal artery scanning falls short as a secondary, noninvasive thermometry method for trauma patients. AB - Oral and mandibular trauma pose barriers to oral thermometry. We sought to determine whether temporal artery (TA) scanning thermometry could be an accurate, noninvasive back up method of thermometry in patients with these types of traumatic injury. We compared 3 techniques of TA scanning, axillary thermometry, and oral thermometry in critical care patients. Our results indicate that TA scanning methods were, at best, comparable with axillary measurements. In addition, the performance of the TA scanners varied with body mass index, whereas axillary readings did not. PMID- 19305300 TI - Therapeutic hypothermia in the postresuscitation patient: the development and implementation of an evidence-based protocol for the emergency department. AB - Studies have shown that therapeutic hypothermia (TH) improves outcomes in patients who have experienced a cardiac arrest (; ). This article discusses TH and the process used by one emergency department to develop and implement an evidence-based protocol on TH for the postresuscitation patient. PMID- 19305302 TI - From the editor. Leadership in nursing administration. PMID- 19305303 TI - The never-ending road to excellence. PMID- 19305304 TI - Living excellence: life after Magnet designation. AB - The achievement of Magnet recognition is the beginning of a new way of being as an organization. Strategies to support innovation leadership, value-based decision making, agility, sustainability of excellence, technology advancements, and lifelong learning are discussed within the framework of the Magnet organization. Behaviors and challenges of living the expectations of the Magnet organization are presented as opportunities to assist healthcare leaders in this important work. PMID- 19305305 TI - Bumps on the road to Magnet designation: achieving organizational excellence. AB - The chief nursing officer is in a unique position to guide his or her organization to excellence by creating a compelling vision; maintaining objectivity regarding the nursing department's accomplishments; holding senior nurse leaders accountable as Magnet champions; demonstrating strategic thinking, business planning development, operational connection, and awareness of clinical aspects of care; and establishing levels of ownership and decision making within the nursing department's operational framework. The clear definition of terms including responsibility, authority, delegation, accountability, and empowerment are necessary and, coupled with specific actions, skills, and measures of success, guide individual and group processes to achieve organizational excellence and ultimately Magnet designation. PMID- 19305306 TI - Administration and union partnership: one Magnet hospital's story. AB - For each hospital that has achieved Magnet designation as an organization of excellence, a unique story exists about its experience. Our story of excellence is founded in a long partnership between administration and union leadership spanning 2 decades. We recognize that excellence is not a steady state, rather a continued commitment to improved patient, staff, and organizational outcomes. This article provides a glimpse into one hospital's story-administration and union partnership in achieving and sustaining the American Nurses Credentialing Center's Magnet designation. PMID- 19305307 TI - Managing the complexity of a systemwide electronic medical record design and implementation: lessons for nurse leaders. AB - Nurse executives are critical to the successful planning, implementation, and adoption of technology to support the workflow and documentation needs of nurses and other end users. Informed by key principles of complexity theory, the enormous task of broad-based implementation of a standardized electronic medical record can be accomplished through the thoughtful development of flexible structures and change management principles that promote intelligent decision making and adoption by key stakeholders. PMID- 19305308 TI - Effects of organizational change on work-related empowerment, employee satisfaction, and motivation. AB - This article reports the results of a longitudinal quantitative study on nurses' views on factors promoting and impeding empowerment and examines the relationship between work-related empowerment and background variables in one hospital. Data were collected using a self-administered questionnaire and analyzed statistically. Nurses gave lowest assessments of promoting factors on the second measurement occasion, a time when the organization was going through major changes. Both job satisfaction and motivation showed a positive relationship with factors promoting empowerment. Organizational changes have a direct effect on the work environment in terms of empowerment and job satisfaction. To cope successfully with changes, special attention must be paid to personnel management. It seems that factors promoting and impeding empowerment can be used to measure effects of organizational changes as well. PMID- 19305309 TI - Transition from leadership development to succession management. AB - As nurse executives approach retirement, it is imperative to conduct succession management for our replacements, along with other key nursing leadership positions. Nurse executives are professionally obligated to consciously and strategically develop leaders. Start by identifying key positions. Conduct a migration risk assessment. Establish core competencies for executive-, director-, and manager-level positions. Identify top performers and conduct individual needs assessments. Develop individualized progressive development plans and identify associated critical success factors. Prioritize, coach, and mentor top performers. This article describes the implementation of a succession management plan and provides a framework for transitioning from leadership development to succession management. PMID- 19305310 TI - Challenges in sustaining excellence over time. AB - A culture of excellence is not synonymous with a culture of perfection. Perfection is not attainable but excellence is when an infrastructure is developed that can rapidly and effectively adapt to change. Saint Joseph's Hospital in Atlanta, Georgia, is often asked about the challenges faced in developing, and sustaining, a culture of nursing excellence over time. Topics such as organizational hardiness, the emergence of a professional practice model, the development of a clinical nurse advancement program, and the implementation of the Forces of Magnetism and the Essentials of Magnetism will be discussed in relation to sustaining a culture of excellence. PMID- 19305311 TI - On the scene: St Mary's Hospital, Madison, Wisconsin. AB - In this article, we discuss Shared Governance as the foundation of our nursing professional practice model. Through the use of case examples and reflections from our management team, we demonstrate how this accountability-based practice model promotes excellence through developing, connecting, and engaging people, clarifying and communicating goals, using data to make decisions, and even shaping our organizational response to a critical incident. We close with a look to our future as our hospital embraces whole-system shared decision making. PMID- 19305312 TI - The CNO: challenges and opportunities on the journey to excellence. AB - Moving a southern rural community medical center through the multitude of regulatory, fiscal, social, cultural, and political changes required for healthcare excellence in the early 21st century has provided some particularly unique challenges. This article describes opportunities and initiatives taken by a chief nursing officer and key members of the healthcare team to meet the challenges of moving a rural and rapidly growing healthcare system toward excellence. The initiatives and perspectives of the team, the chief nursing officer, performance improvement/quality corporate director, the board chairman, and nursing leaders, are presented. PMID- 19305313 TI - Evidence and leadership. AB - The call for evidence has swept the healthcare landscape in medicine and nursing and more recently in nursing management. Nurse managers and administrators have been identified as important to support research use and evidence-based practice. The primary focus of this article is (1) to describe the concept of evidence, (2) to challenge the wisdom of basing the practice of leadership on a narrow (reductionistic) understanding of the concept of evidence, and (3) to argue for a rethinking on what should count as evidence in nursing administration. Semantic analysis and examination of dictionaries reveal that evidence as a concept has both an internal and an external dimension. To understand the concept of evidence in a profound sense as well as its significance for leadership this article describes 3 different meanings of the concept: ontological, contextual, and King's evidence. PMID- 19305314 TI - Incentives and procedures effective in increasing survey participation of professional nurses in hospitals. AB - Positive patient and nurse outcomes depend upon improved clinical nurse work environments. Improvements can be confirmed through environmental surveys that have sufficient participation to yield accurate data. Professional clinical nurses in hospitals face a unique work complexity that inhibits survey participation. What procedures/incentives are effective in increasing survey participation? Are procedures/incentives effective with non-nurses applicable to nurses? These questions were answered through a focused literature review, a case study analysis of procedures and incentives used by M.D. Anderson Cancer Center that resulted in a 93% response rate on an environmental Web-based survey, operational data from environmental surveys administered to staff nurses in 286 hospitals nationally and internationally, and an e-mail survey of nurse leaders/managers in 7 strategically selected hospital units. Incentives effective with nurse populations differed from those of non-nurse populations in several areas. Benchmarking and gap analysis-assessment, comparison to internal or external referents, introduction of needed improvements, and reassessment-are best leadership/management practices and are essential elements of the research and quality improvement efforts characteristic of Magnet hospitals. As the use of surveys for quality improvement and workforce management increases, nurse leaders need to know what procedures and incentives encourage response rates large enough to produce valid, representative data. PMID- 19305315 TI - Boards of nursing commitment to ongoing regulatory excellence. PMID- 19305317 TI - Preterm birth: an enigma and a priority. PMID- 19305318 TI - Ovarian cancer: to screen or not to screen. PMID- 19305319 TI - Results from four rounds of ovarian cancer screening in a randomized trial. AB - OBJECTIVE: To test whether annual screening with transvaginal ultrasonography and CA 125 reduces ovarian cancer mortality. METHODS: Data from the first four annual screens, denoted T0-T3, are reported. A CA 125 value at or above 35 units/mL or an abnormality on transvaginal ultrasonography was considered a positive screen. Diagnostic follow-up of positive screens was performed at the discretion of participants' physicians. Diagnostic procedures and cancers were tracked and verified through medical records. RESULTS: Among 34,261 screening arm women without prior oophorectomy, compliance with screening ranged from 83.1% (T0) to 77.6% (T3). Screen positivity rates declined slightly with transvaginal ultrasonography, from 4.6 at T0 to 2.9-3.4 at T1-T3; CA 125 positivity rates (range 1.4-1.8%) showed no time trend. Eighty-nine invasive ovarian or peritoneal cancers were diagnosed; 60 were screen detected. The positive predictive value (PPV) and cancer yield per 10,000 women screened on the combination of tests were similar across screening rounds (range 1.0-1.3% for PPV and 4.7-6.2 for yield); however, the biopsy (surgery) rate among screen positives decreased from 34% at T0 to 15-20% at T1-T3. The overall ratio of surgeries to screen-detected cancers was 19.5:1. Seventy-two percent of screen-detected cases were late stage (III/IV). CONCLUSION: Through four screening rounds, the ratio of surgeries to screen-detected cancers was high, and most cases were late stage. However, the effect of screening on mortality is as yet unknown. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00002540 LEVEL OF EVIDENCE: II. PMID- 19305320 TI - Synchronous primary ovarian and endometrial cancers: a population-based assessment of survival. AB - OBJECTIVE: To estimate the occurrence of synchronous epithelial ovarian and endometrial cancers among ovarian cancer patients and to assess survival of women with synchronous cancers. METHODS: Synchronous ovarian and endometrial cases were identified using data from the Surveillance, Epidemiology, and End Results Program from 1973 to 2005. Multivariable Cox-proportional hazards regression was used to estimate risk of death from ovarian cancer, comparing synchronous ovarian and endometrial cancers with single ovarian cancers and adjusting for demographic, prognostic, and treatment characteristics. RESULTS: Synchronous cancers represented less than 3% of the 56,986 epithelial ovarian cancer cases, regardless of the time interval between detection of both cancers. Favorable characteristics for synchronous patients included younger age at diagnosis, earlier stage of disease, and better grade of disease. Multivariable adjusted hazard ratios showed a 25% reduction in risk of death from ovarian cancer for synchronous tumors compared with single tumors (hazard ratio 0.75, 95% confidence interval [CI] 0.66-0.85). In stratified analysis by stage, the corresponding hazard ratios for localized and distant stages were 0.63 (95% CI 0.42-0.95) and 0.70 (95% CI 0.60-0.81), respectively. CONCLUSION: Women with synchronous ovarian and endometrial cancers exhibit favorable survival outcomes as compared with patients with single ovarian cancers, even after adjusting for demographic, prognostic (including stage), and treatment characteristics. The survival advantage associated with having synchronous cancers persisted after analyses were restricted to distant stage. LEVEL OF EVIDENCE: III. PMID- 19305321 TI - Secondary cancer prevention during follow-up for endometrial cancer. AB - OBJECTIVE: To conduct a population-based study of incidence and screening rates for secondary breast and colorectal cancers after endometrial cancer and to assess implications for follow-up. METHODS: This was a retrospective population based study from administrative databases. The inception cohort included all women diagnosed with endometrial cancer in Ontario, Canada from 1996 to 2000, without a previous history of breast or colorectal cancer. We ascertained 5-year recurrence and overall survival rates and practitioner type during follow-up. Primary outcomes were age-standardized incidence and screening rates of breast and colorectal cancer during follow-up compared with the general female population. RESULTS: There were 3,473 women in the cohort. The 5-year recurrence rate was 15.0% and overall survival was 79.3%. Family physicians were most often involved in follow-up care. Age-standardized incidence rates of breast and colorectal cancer were 0.5% and 0.7%, respectively, compared with 0.5% (P=.76) and 0.2% (P<.001) in the general population. Age-standardized screening rates for these cancers were 64.0% and 30.0%, respectively, compared with 31.0% (P<.008) and 15.0% (P<.001) in the general population. Women aged older than 70 years and those with the lowest income were least likely to have secondary cancer screening. CONCLUSION: Women with endometrial cancer have a comparable risk of breast cancer but higher risk of colorectal cancer compared with the general population. Follow-up after endometrial cancer should include counseling and uptake of secondary cancer prevention strategies, which will contribute to maximizing long-term survivorship for these women. LEVEL OF EVIDENCE: II. PMID- 19305322 TI - Labor induction process improvement: a patient quality-of-care initiative. AB - OBJECTIVE: To examine the effects that medical staff education and a new process for scheduling inductions had on decreasing inappropriate inductions. METHODS: At our institution in 2004, guidelines were developed and shared with the medical staff and reinforced in 2005. The guidelines for elective induction required patients to have completed 39 weeks of gestation and to have a Bishop score of at least 8 for nulliparas and 6 for multiparas. In 2006, the induction scheduling process was changed and the guidelines were strictly enforced. All scheduled inductions during the same 3-month time period (June through August) in 2004 (n=533) and 2005 (n=454) and during a 13-month period from November 2006 to December 2007 (n=1,806) were compared. Outcomes included elective inductions less than 39 weeks, cesarean birth rate for elective inductions among nulliparas, and the overall induction rate. RESULTS: From 2004-2007, the overall induction rate dropped from 24.9% to 16.6%, a 33% reduction(P<.001); the elective induction rate dropped from 9.1% to 6.4%, a 30% reduction (P<.001); the percentage of elective inductions before 39 weeks of gestation dropped from 11.8% to 4.3%, a decrease of 64% (P<.001); and the frequency of cesarean delivery among nulliparas undergoing elective induction dropped from 34.5% to 13.8%, a decrease of 60%. (P=.01). CONCLUSION: Medical staff education and the development and enforcement of induction guidelines contributed to a decrease in inappropriate inductions, a lower cesarean birth rate for electively induced nulliparas, and a lower elective and overall induction rate. LEVEL OF EVIDENCE: III. PMID- 19305323 TI - Decreasing elective deliveries before 39 weeks of gestation in an integrated health care system. AB - OBJECTIVE: The American College of Obstetricians and Gynecologists has recommended that elective deliveries not be performed before 39 weeks of gestation, to minimize prematurity-related neonatal complications. Because a worrisome number of elective deliveries were occurring before 39 weeks of gestation in our system, we developed and implemented a program to decrease the number of these early term elective deliveries. Secondary objectives were to monitor relevant clinical outcomes. METHODS: The electronic medical records of an integrated health care system involving nine labor and delivery units in Utah were queried to establish the incidence of patients admitted for elective induction of labor or planned elective cesarean delivery. These facilities have open staff models with obstetricians, family practitioners, and certified nurse midwives. Guidelines were developed and implemented to discourage early term elective deliveries. The prevalence of early term elective deliveries was tracked and reported back regularly to the obstetric leadership and obstetric departments at each facility. RESULTS: The baseline prevalence of early term elective deliveries was 28% of all elective deliveries before the initiation of the program. Within 6 months of initiating the program, the incidence of near-term elective deliveries decreased to less than 10% and after 6 years continues to be less than 3%. A reduced length of stay in labor and delivery occurred with the introduction of the program, and there were no adverse effects on secondary clinical outcomes. CONCLUSION: With institutional commitment, it is possible to substantially reduce and sustain a decline in the incidence of elective deliveries before 39 weeks of gestation. LEVEL OF EVIDENCE: III. PMID- 19305324 TI - The prognosis for spontaneous labor in women with uncomplicated term pregnancies: implications for cesarean delivery on maternal request. AB - OBJECTIVE: To assess the prognosis for vaginal delivery in women with entirely normal pregnancies who began spontaneous labor at term. METHODS: Between January 1, 1988, and October 31, 2006, a total of 278,164 women delivered newborns at our hospital. A subset of women with uncomplicated pregnancies and spontaneous labor between 37 and 41 weeks of gestation then were identified for analysis of maternal and neonatal outcomes. The outcomes we studied included admission-to delivery intervals, use of epidural analgesia, maternal perineal trauma, route of delivery, and several potential indices of neonatal condition at birth. RESULTS: There were 103,526 (37%) women who delivered at our hospital during the study period who had normal term pregnancies and entered labor spontaneously. Overall, 96% of these women had vaginal deliveries, and adverse neonatal outcomes were rare. For example, perinatal deaths occurred in 0.3 of every 1,000 women. CONCLUSION: Approximately one third of pregnant women have entirely normal pregnancies and enter spontaneous labor at term. Virtually all such women can anticipate safe vaginal deliveries for themselves and their infants. LEVEL OF EVIDENCE: III. PMID- 19305325 TI - Interpregnancy weight gain and cesarean delivery risk in women with a history of gestational diabetes. AB - OBJECTIVE: Along with the rising prevalence of obesity, rates of gestational diabetes mellitus (GDM) and associated adverse outcomes also have increased. We conducted a population-based, retrospective cohort study to assess the association of weight gain between pregnancies with cesarean delivery for the subsequent pregnancy among women with a history of GDM. METHODS: Using linked birth-certificate data for women with at least two singleton births in Washington State during the period from 1992-2005, we identified 2,753 women with GDM who delivered vaginally at the baseline pregnancy (first pregnancy on record). The interpregnancy weight change (subsequent-baseline prepregnancy weight) for each woman was calculated and assigned to one of three categories: weight loss (more than 10 lb), weight stable (+/-10 lb), or weight gain (more than 10 lb). Multiple logistic regression was used to calculate the risk (odds ratio [OR]) of cesarean delivery at the subsequent pregnancy among the weight-gain and weight-loss groups relative to the weight-stable category. RESULTS: Among 2,581 eligible women, 10.9% lost more than 10 lb between pregnancies, 54.0% were weight-stable, and 35.1% gained more than 10 lb. Women who gained more than 10 lb had an adjusted OR for subsequent cesarean delivery of 1.70 (95% confidence interval [CI] 1.16-2.49, 9.7% of women who gained weight), whereas the adjusted OR for women who lost weight was 0.55 (95% CI 0.28-1.10, 4.7% of women who lost weight). CONCLUSION: Women with a history of GDM who gained more than 10 lb between pregnancies are at increased risk of future cesarean delivery. Appropriate weight management among women with a history of GDM may result in decreased cesarean delivery rates along with decreases in associated excess risks and costs. LEVEL OF EVIDENCE: II. PMID- 19305326 TI - The tampon test for vulvodynia treatment outcomes research: reliability, construct validity, and responsiveness. AB - OBJECTIVE: A standardized tampon insertion and removal test, the Tampon Test provides an alternative to sexual intercourse pain as an outcome measure for vulvodynia research. We report upon the reliability, validity, and responsiveness to change of the Tampon Test as an outcome measure for vulvodynia clinical trials. METHODS: Outcome measures were assessed in women enrolled in the Vulvar Vestibulitis Clinical Trial, a randomized clinical trial of oral desipramine and topical lidocaine effectiveness. Reliability estimates of the Tampon Test using the Kappa statistic evaluated week-to-week measures at baseline. Tampon Test construct and discriminant validity were assessed through correlation with other outcome measures. Patients' ability to regularly perform the Tampon Test was compared with regularity of reporting intercourse pain. RESULTS: During the 2 week baseline phase, women with vulvodynia reported stable mean Tampon Test scores 4.6+/-2.6 (week -2); 4.6+/-2.7 (week -1); and 4.7+/-2.8 (week 0) with moderate week-to-week reliability (weighted Kappa 0.52). Over an 8-week phase of trial intervention, change in the Tampon Test measure significantly correlated to a number of outcome measures, including daily pain (r=0.42), intercourse pain (r=0.35), cotton swab vestibular pain (r=0.38), and the Brief Pain Inventory (r=0.49). Women with vulvodynia study participants performed the Tampon Test 96.3% of the requested time, which was twofold higher adherence than intercourse pain measurement (49.7%). CONCLUSION: The Tampon Test reflects a real life experience that is reliable, with good construct validity as shown by the breadth of correlated outcome measures. The Tampon Test is an appropriate outcome measure for vulvodynia research that can be considered for use as the primary efficacy endpoint in clinical trials of treatments for vulvodynia. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00276068 LEVEL OF EVIDENCE: II. PMID- 19305327 TI - Interest in intrauterine contraception among seekers of emergency contraception and pregnancy testing. AB - OBJECTIVE: To estimate the interest in using intrauterine contraception among women and adolescent girls seeking emergency contraception or walk-in pregnancy testing. METHODS: We surveyed 412 women and adolescent girls who requested emergency contraception or pregnancy testing at four family planning clinics in Pittsburgh, Pennsylvania. The 41-item survey assessed knowledge of, attitudes toward, and interest in using an intrauterine device (IUD). Data were analyzed using chi2 and Fisher exact tests and multivariable logistic regression methods. RESULTS: The response rate was 85%. Twelve percent (95% confidence interval [CI] 9-15) of women and adolescent girls surveyed expressed interest in same-day insertion of an IUD, and 22% (95% CI 18-26) wanted more information about IUDs. Interest in same-day IUD insertion increased with higher education level, prior unwanted pregnancy, and experience with barriers to use of contraception. CONCLUSION: Same-day IUD insertion may be a reasonable way to increase the use of highly-effective contraception among women and adolescent girls seeking emergency contraception or walk-in pregnancy testing. LEVEL OF EVIDENCE: II. PMID- 19305328 TI - Adolescent comprehension of emergency contraception in New York City. AB - OBJECTIVE: To estimate comprehension of the over-the-counter emergency contraception label among female adolescents aged 12 through 17 years, and to compare the results with a similar study that focused on adults. METHODS: Surveys were administered to female adolescents in New York City in public venues such as malls, movie theaters, and parks. Participants were asked to read the emergency contraception (levonorgestrel) label before answering survey questions. Comparisons were made in SPSS version 13.0 using chi tests of independence and Fisher exact tests for sparse data. RESULTS: One thousand eighty-five girls between the ages of 12 and 17 participated in the study. Overall, adolescents demonstrated high comprehension of the key points of emergency contraception: (1) that it is a method of preventing pregnancy 92% (confidence interval [CI] 91 94%); (2) that it has to be taken within the first 72 hours after unprotected intercourse 83% (CI 83-87%); (3) that if you are already pregnant emergency contraception will not be effective 87% (CI 85-89%); (4) that emergency contraception will not protect against human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) 95% (CI 94-96%); and (5) that emergency contraception should not be used as a method of long-term birth control 85% (CI 83-87%). CONCLUSION: After reading the emergency contraception (levonorgestrel) label, female adolescents aged 12 to 17 understood the information necessary to use emergency contraception safely and effectively as well as their adult counterparts. LEVEL OF EVIDENCE: III. PMID- 19305329 TI - Frequency and risk factors for repeat abortions after surgical compared with medical termination of pregnancy. AB - OBJECTIVE: To compare the frequency and risk factors for repeat abortions after surgical compared with medical termination of pregnancy. METHODS: Frequency of and risk factors for repeat abortions after medical (performed with mifepristone alone, or with a combination of mifepristone and misoprostol or other prostaglandins) compared with surgical (dilation and curettage, or vacuum aspiration) termination of pregnancy were studied using Finnish national health registries. The cohort consisted of 40,360 women undergoing termination of pregnancy between 2000 and 2005 (19,841 medical and 20,519 surgical abortions) with duration of gestation of 63 days or less. Univariable and multivariable association models were used in connection with various factors associated with repeat abortion. The mean (+/-standard deviation) follow-up times were 3.0 (+/ 1.5) and 4.3 (+/-1.9) years, respectively. RESULTS: Women choosing surgical and medical abortion differed subtly, but significantly in several respects. The total number of repeat terminations was 37.9 per follow-up year per 1,000 after surgical termination of pregnancy and 40.4 after medical termination of pregnancy (P=.01). However, medical termination of pregnancy was not linked to an increased risk of another abortion when compared with surgical methods (hazard ratio [HR] 0.98, 95% confidence interval [CI] 0.93-1.04). In multivariable analysis, the risk factors for repeat abortion were parity (HR 1.99, 95% CI 1.85-2.14), previous abortion(s) (HR 1.70, 95% CI 1.60-1.82), low socioeconomic status (HR 1.22, 95% CI 1.06-1.39), and being unmarried but cohabiting (HR 1.14, 95% CI 1.03 1.25) or single (HR 1.25, 95% CI 1.15-1.36). The risk of repeat termination of pregnancy decreased with age, among women living in rural areas, and when intrauterine devices or sterilization were planned for future contraception. CONCLUSION: The risk of repeat abortion is associated with various sociodemographic characteristics. The method of abortion used is not a risk factor for repeat termination of pregnancy. LEVEL OF EVIDENCE: II. PMID- 19305330 TI - Remote hemodynamics and renal function in formerly preeclamptic women. AB - OBJECTIVE: Women with a history of preeclampsia have an increased risk of developing chronic hypertension and cardiovascular disease. However, little is known about the mechanism responsible for vascular disease in formerly preeclamptic women. The aim of our study was to test whether preeclampsia predisposes to central hemodynamic and renal impairments 20 years after pregnancy. METHODS: In this cross-sectional study, 22 formerly preeclamptic women and 29 parous controls participated, matched for body mass index, age, and date of birth. All women delivered in the period of 1979-1987. Measures included automated blood pressure, Doppler echocardiography, microalbuminuria, paraaminohippurate, and creatinine clearances. Hypertension was defined as blood pressure at or above 140/90 mm Hg, using antihypertensive drugs, or both. RESULTS: Hypertension was present in 55% of the formerly preeclamptic women and 7% of the women in the control group (P<.01). Mean arterial pressure was higher in the formerly preeclamptic women compared with those in the control group (100 and 88 mm Hg, respectively, P<.01). Peripheral vascular resistance was about 20% higher, renal vascular resistance about 30% higher, and renal blood flow about 15% lower in the formerly preeclamptic women compared with those in the control group (P<.05). Similar results were observed after stratification for hypertension in both groups. CONCLUSION: Both normotensive and hypertensive middle-aged, formerly preeclamptic women showed impaired central hemodynamic and renal function compared with parous controls. LEVEL OF EVIDENCE: II. PMID- 19305331 TI - Ductus venosus Doppler at 11 to 13 weeks of gestation in the prediction of outcome in twin pregnancies. AB - OBJECTIVE: To examine the independent contribution of abnormal flow in the ductus venosus at 11 to 13 weeks of gestation in the prediction of adverse pregnancy outcome in relation to chorionicity. METHODS: This was a prospective study in 516 dichorionic and 179 monochorionic twin pregnancies in which the fetal ductus venosus flow was assessed at 11 0/7 to 13 6/7 weeks of gestation. The prevalence of reversed a-wave in the fetal ductus venosus was compared between monochorionic and dichorionic pregnancies and between those with and without pregnancy complications. Comparisons between each of the pregnancy outcomes and the normal outcome group and between monochorionic and dichorionic pregnancies were made using the Mann-Whitney U-test for continuous variables and the chi2 test and Fisher exact test for categorical variables. RESULTS: The prevalence of reversed a-wave in at least one of the fetuses was significantly higher in monochorionic than in dichorionic pregnancies (18.4% compared with 8.3%, P<.001) and in pregnancies complicated by miscarriage (28.6%, P=.005), fetal aneuploidy (70.0%, P<.001), and twin-twin transfusion syndrome (38.5%, P<.001) compared with the pregnancies with two healthy live births (7.7%). Pregnancy outcome was normal in 33 of the 43 (76.7%) dichorionic and in 14 of the 33 (42.4%) monochorionic twins with reversed a-wave in at least one of the fetuses. CONCLUSION: In twins, reversed a-wave in the ductus venosus at 11 to 13 weeks of gestation is associated with increased risk for aneuploidies, miscarriage, and development of severe twin-twin transfusion syndrome. However, in about 75% of dichorionic twins and 40% of monochorionic twins with reversed a-wave, the pregnancy outcome is normal. LEVEL OF EVIDENCE: II. PMID- 19305332 TI - Prevalence of domestic violence in a pregnant military population. AB - OBJECTIVE: To estimate the prevalence of domestic violence and the characteristics of pregnant women reporting domestic violence in a military setting. METHODS: This was a prospective observational study of patients presenting for prenatal care to a Naval hospital from January 2007 to March 2008. Participants were screened anonymously for domestic violence using the Abuse Assessment Screen. Data were summarized using medians, interquartile ranges, and frequency distributions. Univariable comparisons between groups were conducted using Mann-Whitney tests for continuous data and chi2 tests for categorical outcomes RESULTS: Of the 1,162 surveys, 14.5% screened positive for abuse (either current or past), and 1.5% of respondents reported current pregnancy abuse. Relative to married women, single women (odds ratio [OR] 1.81, 95% confidence interval [CI] 1.04-3.16, P=.036) and separated or divorced women (OR 3.45, 95% CI 1.59-7.46, P=.002) were at an overall increased risk of physical or emotional partner abuse. Compared with married women, the single women (OR 2.80, 95% CI 1.35-5.78, P=.005), but not the separated or divorced women (P=.172), were at increased risk for partner abuse in the previous 12 months. A family history of abuse also was associated with an increased risk of abuse within the previous 12 months (OR 5.99, 95% CI 2.99-11.99, P<.001). CONCLUSION: The prevalence of domestic violence in our pregnant military population was 14.5%, which is in the upper range of the prevalence reported in a nonmilitary population (0.9-23%). Unmarried status and a history of abuse may indicate a higher abuse risk. LEVEL OF EVIDENCE: III. PMID- 19305333 TI - Biomechanical analyses of the efficacy of patterns of maternal effort on second stage progress. AB - OBJECTIVE: To develop and use a biomechanical computer model to simulate the effect of varying the timing of voluntary maternal pushes during uterine contraction on second-stage labor duration. METHODS: Published initial pelvic floor geometry was imported into technical computing software to build a simplified three-dimensional biomechanical model with six representative viscoelastic levator muscle bands interconnected by a hyperelastic iliococcygeal raphe. An incompressible sphere simulated the molded fetal head. Forces from uterine contraction and voluntary expulsive efforts were summed to push the model fetal head along the curve of Carus opposed by the resistance of the pelvic floor structures to stretch. Holding uterine maximal contraction force and push strength constant, pushes were timed before ("pre"), at ("peak"), and after ("post") maximal uterine contraction force. The effect of different combinations of pushes on second stage duration and the number of pushes required for delivery were evaluated. RESULTS: Calculated second stage durations ranged from 57.5 minutes (triple or pre-peak-post pattern) to 75.8 minutes (prepush and postpush patterns). Delivery with the triple-push pattern required 59 voluntary pushes, while the peak-push pattern required 23 voluntary pushes, a 61% reduction. The corresponding reduction for the pre-and-peak-push pattern was 29%, the peak-and post push pattern was 30%, the prepush pattern was 54%, and the postpush pattern was 56%. CONCLUSION: Although the triple-push pattern resulted in a 16% shorter second stage, this came at the energetic expense of a 61% increase in the number of pushes required. LEVEL OF EVIDENCE: III. PMID- 19305334 TI - Saline-infusion sonography endometrial sampling compared with endometrial biopsy in diagnosing endometrial pathology. AB - OBJECTIVE: To evaluate prospectively whether saline-infusion sonography endometrial sampling will improve diagnosis of benign and malignant endometrial disease compared with blind endometrial biopsy in perimenopausal and postmenopausal women with abnormal uterine bleeding. METHODS: After initial assessments and endometrial biopsies, women aged 40 or older with abnormal uterine bleeding were referred for transvaginal ultrasonography. Patients with abnormal endometrial evaluations were offered sonohysterography and invited to enter the saline-infusion sonography endometrial sampling study. Saline-infusion sonography endometrial sampling was performed at the end of sonohysterography using an endometrial sampling curette at the sight of the endometrial abnormality or at a representative site in the endometrial cavity if the cavity was normal. Histopathologic diagnoses of blind endometrial biopsy and saline-infusion sonography endometrial sampling were compared with final outcomes. Kappa values for the two sampling techniques were calculated for diagnostic accuracy. RESULTS: A total of 88 saline-infusion sonography endometrial samples were obtained; final outcomes were attained in 80 cases. When comparing saline-infusion sonography endometrial sampling with final outcomes, saline-infusion sonography endometrial sampling provided a diagnosis 89% of the time (95% confidence interval [CI] 82 95%) compared with endometrial biopsy at 52% (95% CI 42-62%). Forty-five patients with endometrial biopsies had final outcomes. When comparing endometrial biopsy with final outcomes, endometrial biopsy underestimated the incidence of pathology, especially when focal lesions and malignancy were present. This difference was statistically significant (P<.005). CONCLUSION: Saline-infusion sonography endometrial sampling is superior to endometrial biopsy in diagnosing endometrial pathology in perimenopausal and postmenopausal women with abnormal uterine bleeding. LEVEL OF EVIDENCE: II. PMID- 19305335 TI - Clinical significance of cytologic atypical squamous cells of undetermined significance. AB - OBJECTIVE: To analyze the screening status, clinical characteristics, and risk of invasive cervical cancer in women with cytologic atypical squamous cells of undetermined significance (ASC-US) in general populations and to make recommendations for these women. METHODS: The 95,309 women with initial cytologic ASC-US from the national cervical cancer screening database in Taiwan were divided into unscreened and screened groups based on their previous screening history. Newly diagnosed invasive cervical cancer cases in the follow-up period were retrieved for analysis. RESULTS: There were 860 cases of invasive cervical cancer during the follow-up period of 556,758 person-years, with an overall incidence rate of 154.5 cases per 100,000 person-years. The incidence was 92.3 and 257.2 cases per 100,000 person-years in the screened and unscreened groups, respectively. Previous screening history was a strong risk predictor of invasive cervical cancer (hazard ratio 2.8, 95% confidence interval 2.4-3.1, P<.001) after adjustments for age, educational status, and hospital setting. Age was also a significant risk factor for developing invasive cervical cancer in the unscreened group but not in the screened group. CONCLUSION: Women with cytologic ASC-US, especially those without a previous Pap test or older women, were more likely to develop invasive cervical cancer and should be followed up aggressively. LEVEL OF EVIDENCE: II. PMID- 19305336 TI - Outcome and patterns of recurrence for International Federation of Gynecology and Obstetrics (FIGO) stages I and II squamous cell vulvar cancer. AB - OBJECTIVE: To study patterns of recurrence, to evaluate pathologic features correlating with recurrence, and to estimate the prognostic implications for each different pattern of recurrence in the International Federation of Gynecology and Obstetrics (FIGO) stages I and II squamous cell vulvar cancer. METHODS: This was a retrospective study of 121 cases of vulvar cancer managed at our institution from 1987 to 2005. Time to recurrence, sites of local and distant recurrence, and the type of surgery were recorded. Relapse-free and overall survival were calculated. RESULTS: There was no difference in recurrence rates, time to recurrence, or survival between patients with FIGO stages I or II disease. The 5 year actuarial survival (corrected for competing risks) for stage I disease was 97% compared with 95% for stage II (P=.83). Progression-free survival at 5 years was 86% for stage I and 94% for stage II.In this study, 95.9% of patients were treated with vulvar-conserving surgery without detriment with respect to recurrence or survival. CONCLUSION: Vulvar-conserving surgery, even for large tumors, results in excellent outcomes. Vulvar recurrences have an excellent prognosis, but primary site and remote site vulvar recurrences are biologically different. There is no justification for the FIGO differentiation of node negative cancers confined to the vulva on the basis of tumor size. LEVEL OF EVIDENCE: III. PMID- 19305337 TI - Evidence for a role of hot flushes in vascular function in recently postmenopausal women. AB - OBJECTIVE: Observational studies indicate that postmenopausal hormone therapy (HT) prevents cardiovascular disease, but randomized clinical trials have not confirmed this effect. Hot flushes were more likely to be present in women starting HT in observational studies, whereas these symptoms were mild or absent among women attending randomized clinical trials. We hypothesized that vascular function may differ in women with and without vasomotor hot flushes. METHODS: One hundred forty-three recently postmenopausal women showing a broad range of variation in hot flushes were studied with radial artery tonometry. Pulse wave analyses were assessed at baseline and after nitroglycerin and salbutamol challenges. Wilcoxon signed rank test was used for paired comparisons after challenges with nitroglycerin and salbutamol. RESULTS: Neither baseline arterial stiffness nor endothelial function differed between women without or with mild, moderate, or severe hot flushes. However, after nitroglycerin challenge, the time to the onset of the reflected wave (dependent on pulse wave velocity) was 9.5% longer (P=.014), and the time to the first systolic peak (dependent on the rapid phase of ventricular ejection) was 13.9% longer (P=.025) in women with severe hot flushes as compared with asymptomatic women. CONCLUSION: Women with severe vasomotor hot flushes show greater vascular responsiveness to nitroglycerin than women without hot flushes. This may partially explain the conflicting data between observational and randomized HT studies. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00668603 LEVEL OF EVIDENCE: II. PMID- 19305338 TI - Effects of pregnancy, parturition, and anal sphincter transection on function of the external anal sphincter in an animal model. AB - OBJECTIVE: To estimate the effects of pregnancy, parturition, and anal sphincter laceration (with repair) on external anal sphincter morphology and neurophysiology and to define the time course of these effects after injury. METHODS: Within 4 hours of vaginal delivery, 80 rats underwent either sham or anal sphincter laceration with repair. After 3 days, 3 weeks, and 3 and 6 months (n=20 for each time point), animals were killed, and the anal sphincter complexes dissected and removed for neurophysiologic studies. Twitch tension, peak tetanic force, fatigue, and maximal electrical field-stimulated force generation were determined. Sphincters were then fixed and serially sectioned (5-micrometer thickness) at 100-micrometer intervals for histologic analysis. RESULTS: Maximal electrical field-stimulated force generation, maximal tetanic contraction, and twitch tension were decreased in the external anal sphincter 3 days after anal sphincter laceration with repair compared with sham-operated parturient rats (3.3 g compared with 11.6 g, 4.5 g compared with 14.5 g, and 0.6 g compared with 2.0 g, respectively, all P<.02). Increased fatigability of the sphincter muscle was observed in all newly parturient rats-sham and anal sphincter laceration with repair; recovery occurred in the shams by 3 months. A gradual recovery occurred in all these neurophysiologic measures, with no significant differences between anal sphincter laceration with repair and shams by 6 months postpartum. CONCLUSION: Repaired anal sphincter transection in periparturient animals results in short-term severe compromise of neurophysiologic function of the external anal sphincter. Over time, however, force generation recovers and approximates that of postpartum rats with intact anal sphincters. PMID- 19305339 TI - Human papillomavirus type-distribution in vulvar and vaginal cancers and their associated precursors. AB - OBJECTIVE: Data on human papillomavirus (HPV) prevalence in vulvar and vaginal cancers are limited. These data are important to predict the potential future effect of prophylactic HPV vaccines. Our aim was to conduct a systematic review of HPV type distribution in vulvar and vaginal invasive carcinomas, vulvar intraepithelial neoplasia (VIN), and vaginal intraepithelial neoplasia. DATA SOURCES: A MEDLINE search was conducted using the terms vulvar/vaginal cancer, intraepithelial neoplasia, and HPV/human papillomavirus through September 2007 with no specified start date or language restrictions. METHODS OF STUDY SELECTION: A total of 725 abstracts (564 vulvar, 161 vaginal) were reviewed, of which 67 studies (56 vulvar, 11 vaginal) met the inclusion criteria of using polymerase chain reaction (PCR) or hybrid capture assays for HPV DNA detection and having more than one case with HPV data available. TABULATION, INTEGRATION AND RESULTS: This review identified 2,790 vulvar (1,379 invasive, 1,340 VIN2/3, 71 VIN1) and 315 vaginal cases (83 invasive, 166 vaginal intraepithelial neoplasia 2/3, 66 vaginal intraepithelial neoplasia 1). Most cases were from North America and Europe (87.2%), with few from Asia (5.5%) and South America (7.3%). Human papillomavirus prevalence in vulvar cancer, VIN2/3, and VIN1 was 40.1%, 80.4%, and 77.5%, respectively. HPV prevalence in vaginal cancer, vaginal intraepithelial neoplasia (VAIN)2/3, and VAIN1 was relatively higher at 65.5%, 92.6%, and 98.5%, respectively. HPV16 was the most common type in vulvar (29.3%) and vaginal (55.4%) cancers, VIN2/3 (71.2%) and VAIN2/3 (65.8%). CONCLUSION: Human papillomavirus prevalence was higher among vaginal than vulvar cases, and HPV16 accounted for most HPV-positive cases for both cancers. Although the potential effect of HPV vaccines on these gynecologic cancers may not be as high as for cervical cancer due to their more diverse causes, vaccinating young women against HPV16/18 may help to reduce the incidence of HPV-related cases. PMID- 19305340 TI - Surgeon General's Conference on the Prevention of Preterm Birth. AB - To address the serious and seemingly intractable problem of preterm birth, the Surgeon General's Conference on the Prevention of Preterm Birth convened many of the country's experts from the public and private sectors of research, public health, and health care delivery to discuss preventive strategies. The purpose of the conference was to increase awareness of preterm birth in the United States, review key findings and reports issued by experts in the field, and establish an agenda for activities in both the public and private sectors to mitigate the problem. The six work groups created focused on biomedical research, epidemiological research, psychosocial and behavioral factors in preterm birth, professional education and training, outreach and communication, and quality of care and health services. Several cross-cutting issues between the work groups were identified, and the conference concluded with the request to the Surgeon General to make the prevention of preterm birth a national public health priority. Reaching this goal through the implementation of the conference recommendations will require new resources to create broad-based research capacity, a vigorous national vital records system, multidisciplinary intervention programs, careful study of factors contributing to racial and ethnic disparities, reinvigorated health professional and consumer education programs, and access to high-quality preconception and perinatal healthcare for all Americans. Clinicians must be adequately informed to initiate activities to prevent this serious problem. Recommendations from this conference will inform Congress and create a national agenda to address the identification of the causes, risk factors, prevention, and treatment of preterm birth. PMID- 19305341 TI - When cultures collide: female genital cutting and U.S. obstetric practice. PMID- 19305342 TI - Adolescent gynecology. AB - Given new developments in the field of adolescent reproductive health, this review focuses on highlighting new guidelines and practice patterns in evaluation and management of adolescent gynecologic problems. First, understanding the proper techniques for the initial examination is key to establishing a long-term relationship with this age group. Reservations about the first gynecologic examination are common, and the practitioner's goal is foremost to make the patient as comfortable as possible. Preventive health in this patient population is key, and practitioners should become comfortable with providing education about topics as diverse as sexuality, eating disorders, and dating violence. Furthermore, the frequency with which teenagers report sexual activity and the high unintended pregnancy rate in this age group makes counseling regarding effective contraception essential. Additionally, practitioners are encouraged to take the opportunity to discuss the availability of the human papillomavirus (HPV) vaccine with adolescents. In 2007, adolescents were designated as a special population, given the frequency with which they acquire and clear mild HPV related cervical dysplasia. More conservative treatment in this population is generally favored. During their transition through puberty, disorders of menstruation become the most common complaint requiring the attention of the gynecologist. Most commonly, anovulation serves as the cause behind such abnormal bleeding. Polycystic ovarian syndrome can develop in early puberty and carry its consequences into adulthood. Infertility, diabetes, and hirsutism mark the most important components of the syndrome and require age-appropriate management. Finally, the consequences of endometriosis on the future fertility of adolescents have brought early intervention to light. Recognition and prompt treatment are advocated to prevent the future implications of this disease. PMID- 19305343 TI - Probability of hysterectomy after endometrial ablation. PMID- 19305345 TI - Graft use in transvaginal pelvic organ prolapse repair: a systematic review. PMID- 19305347 TI - ACOG Practice Bulletin No. 103: Hereditary breast and ovarian cancer syndrome. AB - Hereditary breast and ovarian cancer syndrome is an inherited cancer susceptibility syndrome. The hallmarks of this syndrome are multiple family members with breast cancer or ovarian cancer or both, the presence of both breast cancer and ovarian cancer in a single individual, and early age of breast cancer onset. Clinical genetic testing for gene mutations allows physicians to more precisely identify women who are at substantial risk of breast cancer and ovarian cancer. For these individuals, screening and prevention strategies can be instituted to reduce their risks. Obstetricians and gynecologists play an important role in the identification and management of women with hereditary breast and ovarian cancer syndrome. PMID- 19305348 TI - Fungal infections after lung transplantation. AB - PURPOSE OF REVIEW: Lung transplant's Achilles heel is chronic rejection. This is the reason why high immunosuppression is used, which leads to the development of infections. Fungal infections are a great obstacle in lung transplant patients' progress, not only because of their impact on patient survival, but also because fungal infections indirectly have an influence on the graft's progress. This review highlights the changing spectrum of invasive fungal infections as well as the most recent developments in diagnosis, prophylaxis, treatment and monitoring of lung transplantation. RECENT FINDINGS: Fungal infections have a bimodal presentation: early onset, in relation to difficult postsurgeries and prior colonizations, and late onset, primarily in relation to chronic rejection and terminal renal insufficiency. The clinical impact of non-Aspergillus moulds is still unknown. Recent efforts have focused on nonculture-based methods to establish a rapid diagnosis. However, multicentre studies are needed to establish the diagnostic value of galactomannan antigen assay in invasive aspergillosis in lung transplantation. In addition, studies of the sensitivity and specificity of PCR assays are required to establish their diagnostic value. Unfortunately, only some advances in the diagnosis of aspergillosis have been achieved. SUMMARY: Prophylaxis should be tailored according to the different individual patient's risk status. Combined treatments, including surgical therapy, may be useful in some patients. PMID- 19305349 TI - Diagnosis and treatment of lung infection with nontuberculous mycobacteria. AB - PURPOSE OF REVIEW: Pulmonary infections caused by nontuberculous mycobacteria (NTM) are diagnosed with increasing frequency, in part due to growing at-risk populations but also as a result of improved awareness and diagnostic facilities. This review summarizes recent literature regarding epidemiological, clinical, diagnostic and therapeutic aspects of NTM lung infections. RECENT FINDINGS: The number of species known to cause NTM infections has increased due to the extended use of molecular techniques. The number of recognized risk factors, including newly discovered inherited immunological disorders and novel types of immunomodulating drugs such as antagonists of tumor necrosis factor-alpha is also growing. Revised diagnostic criteria for NTM lung infection are available but the decision whether to treat a patient remains a matter of careful individual evaluation taking into account the NTM species, extent of disease, general condition and underlying disorders. No major breakthrough has been made with regard to treatment. Antibiotic treatment of NTM infection is complicated by the necessary long duration and the adverse toxicity profile of many of the potentially effective drugs while there is an uncertain correlation between in vitro susceptibility and in-vivo effectiveness except for two drug-NTM species combinations. The role for novel antibiotics in the treatment of NTM infection is still uncertain. SUMMARY: Much remains unknown regarding treatment of NTM lung infections. In order to provide optimal care, the recommendations provided in the 2007 American Thoracic Society/Infectious Diseases Society of America statement should be taken as a starting point and there should be a low threshold to seek expert consultation. PMID- 19305350 TI - Uterine leiomyomata in relation to insulin-like growth factor-I, insulin, and diabetes. AB - BACKGROUND: Insulin-like growth factor-I (IGF-I) and insulin stimulate cell proliferation in uterine leiomyoma (fibroid) tissue. We hypothesized that circulating levels of these proteins would be associated with increased prevalence and size of uterine fibroids. METHODS: Participants were 35-49-year old, randomly selected members of an urban health plan who were enrolled in the study in 1996-1999. Premenopausal participants were screened for fibroids with ultrasound. Fasting blood samples were collected. Associations between fibroids and diabetes, plasma IGF-I, IGF binding protein 3 (BP3), and insulin were evaluated for blacks (n = 585) and whites (n = 403) by using multiple logistic regression. RESULTS: IGF-I showed no association with fibroids in blacks, but in whites the adjusted odds ratios (aORs) for both mid and upper tertiles compared with the lowest tertile were 0.6 (95% confidence intervals [CI] = 0.3-1.0 and 0.4 1.1, respectively). Insulin and diabetes both tended to be inversely associated with fibroids in blacks. The insulin association was with large fibroids; aOR for the upper insulin tertile relative to the lowest was 0.4 (0.2-0.9). The aOR for diabetes was 0.5 (0.2-1.0). Associations of insulin and diabetes with fibroids were weak for whites. Binding protein 3 showed no association with fibroids. CONCLUSIONS: Contrary to our hypothesis, high circulating IGF-I and insulin were not related to increased fibroid prevalence. Instead, there was suggestion of the opposite. The inverse association with diabetes, although based on small numbers, is consistent with previously reported findings. Future studies might investigate vascular dysfunction as a mediator between hyperinsulinemia or diabetes and possible reduced risk of fibroids. PMID- 19305351 TI - The cost-effectiveness of implementing a new guideline for cardiovascular risk management in primary care in the Netherlands. AB - BACKGROUND: A new Dutch guideline for cardiovascular disease management substantially extends the number of individuals for whom treatment with statins and/or antihypertensive agents is recommended. We estimated the cost effectiveness of implementing the new guideline at the national level. METHODS: First, the number of currently untreated individuals who would become eligible for cholesterol-lowering or antihypertensive treatment under the new guideline was estimated using data from a recent population study. Cost-effectiveness of treating this group of patients was then assessed using a mathematical model. RESULTS: Implementing the guideline in the age category 30-69 years would lead to an additional 465,000 individuals requiring treatment. Over a period of 20 years, the cumulative incidence of acute myocardial infarction in the whole population would drop by 3.0%, that of stroke by 3.9%, and all-cause mortality would drop by 0.9%. The lifetime cost-effectiveness ratio was calculated to be 15,000 Euro per quality-adjusted life year gained. In the age categories 70-79 years and 80 years or above, an additional 600,000 and 450,000 persons, respectively, would need to be treated, resulting in corresponding reductions in cumulative incidences of 14 and 18% (acute myocardial infarction), 17 and 22% (stroke), and 1.2 and 0.6% (all cause mortality) with cost-effectiveness ratios of 20,800 and 32,300 Euro, respectively, per quality-adjusted life year. CONCLUSION: Complete implementation of the new guideline would lead to a considerable increase in the number of individuals requiring treatment. This would be cost-effective up to the age of 70 years. PMID- 19305352 TI - The metabolic syndrome is not a sensible tool for predicting the risk of coronary heart disease. AB - BACKGROUND: The metabolic syndrome (MS) is a popularly used risk marker for coronary heart disease (CHD), yet its utility is in doubt. DESIGN: Cohort study based in Glasgow, Scotland, of 1471 men and women free of cardiovascular disease, followed up for a median of 13.7 years. METHODS: MS was defined according to current criteria, requiring at least three of five dichotomous risk factors to be positive. Cox models were used to obtain hazard ratios (HRs) and discrimination was quantified by areas under receiver operating characteristic curves (AUCs) using 500 bootstrap samples. RESULTS: The HR (95% confidence interval) for CHD, MS versus no MS was 2.23 (1.67-2.97). However, the HR rose monotonically when plotted against the number of positive components, with no suggestion of a threshold effect at three positive components. Furthermore, the HR also changed monotonically as each of the five continuous variables defining the different components increased, again with no obvious threshold effects. The AUC for MS was low, at 0.5764, this being significantly (P<0.0001) lower than the AUCs for other risk prediction models, including the Framingham score, 0.7517. CONCLUSION: Although MS is related to CHD, there is no epidemiological justification for using it, rather than other criteria, as a risk predictor for CHD. PMID- 19305353 TI - Stratification for noninvasive coronary angiography: patient preselection considering atypical angina pectoris, conventional cardiovascular risk assessment, and calcium scoring. AB - BACKGROUND: Noninvasive coronary angiography (NCA) is recommended to be applied as a filter before invasive coronary angiography in patients with intermediate cardiovascular risk. DESIGN: In this prospective single-center study, we validated multislice computed tomography (MSCT) and magnetic resonance imaging (MRI)-based NCA by primarily selecting patients with atypical angina pectoris (AAP). METHODS: In 68 patients (63.6+/-11.4 years) with AAP, the Prospective Cardiovascular Muenster Study score and the Agatston score equivalent (ASE; calcium score) were initially determined for cardiovascular risk assessment. Subsequently, MSCT and MRI-based NCA were performed followed by quantitative invasive coronary angiography for validation. All tests were finally analyzed. RESULTS: CAD (prevalence 38.2%) was diagnosed in 67.7% of patients at high, 61.5% at intermediate, and 37.8% at low cardiovascular risk according to the Prospective Cardiovascular Muenster Study. CAD was diagnosed in 88.2% of patients with an ASE >or=75th percentile and in 90.9% with an ASE >or=90th percentile. With regard to NCA, patient-based analysis resulted in a sensitivity of 96.2% for MSCT versus 69.2% for MRI, in a specificity of 95.2 versus 64.3%, in a positive predictive value of 97.6 versus 77.1%, in a negative predictive value of 92.6 versus 54.5%, and a diagnostic accuracy of 95.6 versus 66.2% (P<0.05). CONCLUSION: In a patient group with a heterogeneous cardiovascular risk, primarily considering AAP allows for effective patient preselection for NCA. This may be optimized by precluding patients with an ASE >or=75th percentile. MSCT rather than MRI may then effectively be involved. PMID- 19305354 TI - Antioxidant activity and phenolic content of Paederia foetida and Syzygium aqueum. AB - The antioxidant activity of fresh and dried plant extracts of Paederia foetida and Syzygium aqueum were studied using beta-carotene bleaching and the 2,2' azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) radical cation assay. The percentage of antioxidant activity for all extract samples using both assays was between 58 and 80%. The fresh samples of both plants had higher antioxidant activity than the dried samples. The results of the beta-carotene bleaching assay were correlated (R(2) = 0.9849) with those of the ABTS assay. PMID- 19305355 TI - Flavonoids with gastroprotective activity. AB - Peptic ulcers are a common disorder of the entire gastrointestinal tract that occurs mainly in the stomach and the proximal duodenum. This disease is multifactorial and its treatment faces great difficulties due to the limited effectiveness and severe side effects of the currently available drugs. The use of natural products for the prevention and treatment of different pathologies is continuously expanding throughout the world. This is particularly true with regards to flavonoids, which represent a highly diverse class of secondary metabolites with potentially beneficial human health effects that is widely distributed in the plant kingdom and currently consumed in large amounts in the diet. They display several pharmacological properties in the gastroprotective area, acting as anti-secretory, cytoprotective and antioxidant agents. Besides their action as gastroprotectives, flavonoids also act in healing of gastric ulcers and additionally these polyphenolic compounds can be new alternatives for suppression or modulation of peptic ulcers associated with H. pylori. In this review, we have summarized the literature on ninety-five flavonoids with varying degrees of antiulcerogenic activity, confirming that flavonoids have a therapeutic potential for the more effective treatment of peptic ulcers. PMID- 19305356 TI - Brominated thiophenes as precursors in the preparation of brominated and arylated anthraquinones. AB - Brominated anthraquinones can be synthesized directly from bromothiophenes when these are reacted with 1,4-naphthoquinones in the presence of meta chloroperoxybenzoic acid. The bromoanthraquinones are versatile building blocks in the preparation of arylated anthraquinones and of extended pi-systems with interspersed anthraquinone units. PMID- 19305357 TI - Antioxidant activities of extracts and main components of Pigeonpea [Cajanus cajan (L.) Millsp.] leaves. AB - Antioxidant activities of the aqueous and ethanol extracts of pigeonpea [Cajanus cajan (L.) Millsp.] leaves, as well as petroleum ether, ethyl acetate, n-butanol and water fractions and the four main compounds separated from the ethanol extract, i.e. cajaninstilbene acid (3-hydroxy-4-prenylmethoxystilbene-2 carboxylic acid), pinostrobin, vitexin and orientin, were examined by a DPPH radical-scavenging assay and a beta-carotene-linoleic acid test. In the DPPH system, the antioxidant activity of the ethanol extracts was superior to that of the aqueous extracts, with IC(50) values were 242.01 and 404.91 microg/mL, respectively. Among the four fractions, the ethyl acetate one showed the highest scavenging activity, with an IC(50) value of 194.98 microg/mL. Cajaninstilbene acid (302.12 microg/mL) and orientin (316.21 microg/mL) showed more efficient radical-scavenging abilities than pinostrobin and vitexin. In the beta-carotene linoleic acid test, the inhibition ratio (%) of the ethyl acetate fraction (94.13%+/-3.41%) was found to be the highest, being almost equal to the inhibition capacity of the positive control BHT (93.89%+/-1.45%) at 4 mg/mL. Pinostrobin (>500 microg/mL) and vitexin (>500 microg/mL) showed insignificant antioxidant activities compared with cajaninstilbene (321.53 microg/mL) and orientin (444.61 microg/mL). In general, the ethyl acetate fraction of the ethanol extract showed greater activity than the main compounds in both systems, such results might be attributed to the synergistic effects of the components. The antioxidant activities of all the tested samples were concentration dependent. Based on the results obtained, we can conclude that the pigeonpea leaf extracts may be valuable natural antioxidant sources and are potentially applicable in both medicine and the healthy food industry. PMID- 19305358 TI - Evaluation of photoluminescence properties of some poly(ethylene glycol) supported coumarin derivatives. AB - The immobilization of some coumarin derivatives on modified poly(ethylene glycol)s is reported and the influence of the polymeric support on the photoluminescence activity of the compounds is discussed. Upon ultraviolet excitation, the derivatives showed coumarin-related emission properties whose peak position and efficiency depended on the loading of the polymer and on the mesomeric effects of the substituents. PMID- 19305359 TI - Efficient one-pot synthesis of indol-3-yl-glycines via uncatalyzed Friedel-Crafts reaction in water. AB - The three component reaction of primary aliphatic amines, glyoxalic acid and indole or N-methylindole in water at ambient temperature affords indol-3-yl or N methylindol-3-yl-glycine in almost quantitative yields. PMID- 19305360 TI - Taiwanese native plants inhibit matrix metalloproteinase-9 activity after ultraviolet B irradiation. AB - Medicinal plants have long been used as a source of therapeutic agents. They are thought to be important anti-aging ingredients in prophylactic medicines. The aim of this study was to screen extracts from Taiwanese plant materials for phenolic contents and measure the corresponding matrix metalloproteinase-9 (MMP-9) activity. We extracted biological ingredients from eight plants native to Taiwan (Alnus formosana, Diospyros discolor, Eriobotrya deflex, Machilus japonica, Pyrrosia polydactylis, Pyrus taiwanensis, Vitis adstricta, Vitis thunbergii). Total phenolic content was measured using the Folin-Ciocalteu method. MMP-9 activities were measured by gelatin zymography. The extracted yields of plants ranged from 3.7 % to 16.9 %. The total phenolic contents ranged from 25.4 to 36.8 mg GAE/g dry material. All of these extracts (except Vitis adstricta Hance) were shown to inhibit MMP-9 activity of WS-1 cell after ultraviolet B irradiation. These findings suggest that total phenolic content may influence MMP-9 activity and that some of the plants with higher phenolic content exhibited various biological activities that could serve as potent inhibitors of the ageing process in the skin. This property might be useful in the production of cosmetics. PMID- 19305361 TI - Anti-hemorrhagic activity of four Brazilian vegetable species against Bothrops jararaca venom. AB - Around 20,000 snakebites are reported annually in Brazil and 90% of them are inflicted by species of the genus Bothrops. Intravenous administration of antibothropic antivenom neutralizes the systemic actions, but it is of little effect on the reversal of local symptoms and often induces adverse reactions, a context that drives the search for complementary treatments for snakebite accidents. Vegetable extracts with a range of antiophidian activities constitute an excellent alternative. In this study, we investigated the anti-hemorrhagic effects of Mouriri pusa Gardn. (Melastomataceae), Byrsonima crassa Niedenzu (Malpighiaceae), Davilla elliptica St. Hill. (Dilleniaceae) and Strychnos pseudoquina St. Hil. (Loganiaceae) against Bothrops jararaca venom. The methanolic extracts from M. pusa (leaves), B. crassa (leaves) and D. elliptica (leaves) showed total neutralization capacity against local hemorrhages. The amenthoflavone and quercetin fractions from B. crassa and the flavonoids fractions (quercetin and myricetin) from M. pusa and D. elliptica also showed total neutralization capacity. We conclude that flavonoids derived from myricetin, quercetin and amenthoflavone play an important role in the anti hemorrhagic potential of these Brazilian vegetables species against B. jararaca venom. PMID- 19305362 TI - Structural changes of malt proteins during boiling. AB - Changes in the physicochemical properties and structure of proteins derived from two malt varieties (Baudin and Guangmai) during wort boiling were investigated by differential scanning calorimetry, SDS-PAGE, two-dimensional electrophoresis, gel filtration chromatography and circular dichroism spectroscopy. The results showed that both protein content and amino acid composition changed only slightly during boiling, and that boiling might cause a gradual unfolding of protein structures, as indicated by the decrease in surface hydrophobicity and free sulfhydryl content and enthalpy value, as well as reduced alpha-helix contents and markedly increased random coil contents. It was also found that major component of both worts was a boiling-resistant protein with a molecular mass of 40 kDa, and that according to the two-dimensional electrophoresis and SE-HPLC analyses, a small amount of soluble aggregates might be formed via hydrophobic interactions. It was thus concluded that changes of protein structure caused by boiling that might influence beer quality are largely independent of malt variety. PMID- 19305364 TI - Laser photolysis and thermolysis of organic selenides and tellurides for chemical gas-phase deposition of nanostructured materials. AB - Laser radiation-induced decomposition of gaseous organic selenides and tellurides resulting in chemical deposition of nanostructured materials on cold surfaces is reviewed with regard to the mechanism of the gas-phase decomposition and properties of the deposited materials. The laser photolysis and laser thermolysis of the Se and Te precursors leading to chalcogen deposition can also serve as a useful approach to nanostructured chalcogen composites and IVA group (Si, Ge, Sn) element chalcogenides provided that it is carried out simultaneously with laser photolysis or thermolysis of polymer and IVA group element precursor. PMID- 19305363 TI - Polyphenols with antiulcerogenic action from aqueous decoction of mango leaves (Mangifera indica L.). AB - This study was designed to determine the gastroprotective effect of a Mangifera indica leaf decoction (AD), on different experimental models in rodents. The administration of AD up to a dose of 5 g/kg (p.o.) did not produce any signs or symptoms of toxicity in the treated animals, while significantly decreasing the severity of gastric damage induced by several gastroprotective models. Oral pre treatment with AD (250, 500 or 1000 mg/kg) in mice and rats with gastric lesions induced by HCl/ethanol, absolute ethanol, non-steroidal anti-inflammatory drug (NSAID) or stress-induced gastric lesions resulted in a significant decrease of said lesions. Phytochemical analyses of AD composition demonstrated the presence of bioactive phenolic compounds that represent 57.3% of total phenolic content in this extract. Two main phenolic compounds were isolated, specifically mangiferin (C-glucopyranoside of 1,3,6,7-tetrahydroxyxanthone) and C-glucosyl-benzophenone (3-C-beta-D-glucopyranosyl-4',2,4,6-tetrahydroxybenzophenone). These findings indicate the potential gastroprotective properties of aqueous decoction from M. indica leaves. PMID- 19305365 TI - Phosphotungstic acid: an efficient, cost-effective and recyclable catalyst for the synthesis of polysubstituted quinolines. AB - Phosphotungstic acid (H(3)PW(12)O(40)) was used as an efficient and recyclable catalyst for the synthesis of polysubstituted quinolines through the Friedlander condensation of 2-aminoarylketone with carbonyl compounds, which was achieved by conventional heating under solvent-free conditions. PMID- 19305366 TI - Ring-substituted 4-hydroxy-1H-quinolin-2-ones: preparation and biological activity. AB - In the study, a series of twelve ring-substituted 4-hydroxy-1H-quinolin-2-one derivatives were prepared. The procedures for synthesis of the compounds are presented. The compounds were analyzed using RP-HPLC to determine lipophilicity and tested for their photosynthesis-inhibiting activity using spinach (Spinacia oleracea L.) chloroplasts. All the synthesized compounds were also evaluated for antifungal activity using in vitro screening with eight fungal strains. For all the compounds, the relationships between the lipophilicity and the chemical structure of the studied compounds are discussed, as well as their structure activity relationships (SAR). PMID- 19305367 TI - Dexlansoprazole (Kapidex) for GERD and erosive esophagitis. PMID- 19305368 TI - Focalin XR for ADHD. PMID- 19305369 TI - In brief: different diets. PMID- 19305370 TI - Adult immunization. AB - Although immunization programs have produced high vaccination rates in US infants and children, similar successes have not been achieved in adults. Vaccines recommended for routine use in adults are reviewed here. Vaccines for travel are reviewed separately. PMID- 19305371 TI - Hepatic stem and progenitor cells in liver diseases and hepatocarcinogenesis. AB - Because of its physiological role and its central function in metabolism and homeostasis, the liver is exposed to an environment rich in toxins. In addition, the liver has to cope with various infectious pathogens, in particular hepatotropic viruses. Therefore, the liver needs efficient and highly regulated regeneration mechanisms. Under normal circumstances the liver shows a low rate of hepatocyte renewal but in the event of liver injury, for example, acute liver damage or drug intoxications, hepatocytes display a remarkable capacity to divide and to restore the liver parenchyma. Because of their enormous capability to regenerate the liver, which is unique among differentiated cells in human organs, hepatocytes function as stem cells. However, if the proliferation of hepatocytes is impaired, as in liver cirrhosis, a progenitor cell population is activated and serves as reserve compartment for liver restoration. Hepatic progenitor cells are bipotential and are located in the canals of Hering, the most peripheral branches of the biliary system. According to the current paradigm, hepatic progenitor cells drive liver regeneration in end-stage liver diseases, where hepatocytes become senescent and may therefore be a target cell population for carcinogenesis. In this review, we revisit landmark studies, summarize the current nomenclature, and discuss recent data elucidating the characteristics and the functional role of hepatic stem and progenitor cells in liver diseases and hepatocarcinogenesis. PMID- 19305372 TI - Epidemiology, screening, diagnosis and treatment of hepatocellular carcinoma. AB - Globally, over half a million people develop hepatocellular carcinoma (HCC) each year and an almost equal number die of it. Some aspects of HCC remain disappointingly unchanged. For example, hepatitis B infection, for which an effective safe vaccine has been developed, remains responsible for a substantial proportion of cases worldwide. Further, the overall survival of patients with HCC remains very low. Nevertheless, the past few years have witnessed several important advances in our understanding of risk factors, screening, as well as treatment of HCC; these advances may change some of the current realities for HCC. In this paper, we will review the epidemiology, screening, diagnosis, and treatment of HCC with special emphasis on recent developments such the role of fatty liver disease, obesity, and coffee may play in causing HCC, the recent guidelines in screening and diagnosis, and state-of-the-art treatment algorithms. PMID- 19305373 TI - Is smoking a prognostic factor in patients with chronic hepatitis C? AB - The progression of chronic liver diseases is characterized by a common histopathological pathway comprising fibrosis formation and distortion of hepatic architecture which are the hallmark of evolution to cirrhosis. Several factors are responsible for the severity and progression of chronic hepatitis C. Here, we describe the most important data regarding the association between regular smoking and histological hepatic lesions. Some reports have shown that the proportion of patients with moderate or significant histological activity gradually increases with the daily consumption of tobacco. Moreover, fibrosis is associated with regular smoking in some studies. However, controversies result from other studies. Nicotine is mainly metabolised by the liver, and its administration in experimental animals showed development of steatosis and focal or confluent hepatic necrosis, probably linked to the oxidative stress associated with lipid peroxidation. In chronic hepatitis C patients, preliminary studies have suggested that hypoxia caused by smoking may induce expression of the cytokines vascular endothelial growth factor (VEGF) and VEGF-D and their corresponding soluble tyrosine kinase receptors fms-like tyrosine kinase receptor and kinase insert domain receptor. Since this issue is controversial and smoking is in any case unsafe, stopping is recommended for patients with liver diseases. PMID- 19305374 TI - Chronic hepatitis C in the advanced adult and elderly subjects. AB - Aging is associated with a complex remodeling of the immune system. While adaptive immune responses show impairment with aging, innate immune responses tend to improve it. Low numbers of CD3+, CD4+ and CD8 T cells have been observed in aged individuals. B lymphocytes tend to diminish as well. However, an increase in NK cells and effector T lymphocytes (CD28- CD8) can be shown. Effector T lymphocytes are characterized by: 1) expression of markers of cytotoxicity; 2) high levels of NK activity; 3) expression of the same inhibitory receptors as NK cells; 4) no cytokine production. For effector T lymphocyte-mediated cytotoxicity of virus-infected cells to occur, viral epitopes need to be exposed on the cell surface in the absence of MCH class I molecule expression, just as it has been shown with NK cells. Indeed, chronic infection with intracellular parasites is known to hinder MHC class I expression on cell surface. In elderly patients with chronic hepatitis C, infected hepatocytes can be shown to express a wide variety of HCV antigens, reflecting latency or active replication, as opposed to low or absent MHC class I expression. This favors elimination of infected hepatocytes by NK cells and effector T lymphocytes. A negative correlation has been observed between outcome of hepatitis and patients' age. Liver biopsies from elderly patients generally show chronic active hepatitis or cirrhosis, which are far less commonly observed in young patients or young adults. Overproduction of proinflammatory cytokines, namely TNF-alpha, IL-1 and IL-6, is responsible for enhanced immuno-phlogosis and underlies a more extensive damage to liver parenchyma. Since interferon-alpha has been shown to upregulate MHC class I molecule expression on infected hepatocytes, it may turn useful as a tool to inhibit NK cell- and effector T lymphocyte-mediated cytotoxicity. Thus, a rationale exists to recommend interferon-a administration in hepatitis C patients, especially in elderly patients. If the data presented here can contribute to foster research into interferon-a treatment of elderly patients with hepatitis C, our goal will be reached. PMID- 19305375 TI - Risk factors for HCV infection. Focus on ethnic and cultural characteristics. AB - A precise understanding of the source of infection and modes of transmission of hepatitis C virus (HCV) is a worldwide priority in terms of public health. This is more evident where multi-ethnic customs cohabit. Despite the knowledge on risk factors for HCV transmission, nearly 50% of infected patients do not have a history suggesting a parenteral route of acquisition. In the present paper, the authors, focusing on ethnic and cultural aspects of HCV transmission, emphasize the need for health education in order to avoid the acquisition and the diffusion of the infection. With the current globalization and large-scale migrations, only by following a preventive strategy based on disseminate information risk behaviours may be modified. PMID- 19305376 TI - Management of primary sclerosing cholangitis. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease with fibrosis surrounding the intrahepatic and/or the extrahepatic bile ducts. PSC is characterized by progressive periductal obliterating fibrosis and bile duct strictures. In individual cases, PSC can have favorable prognosis but in most cases it is a progressive disorder which leads to liver related morbidity, mortality and the need for liver transplantation. In previous early cohorts median survival free of transplantation was 12 years whereas more recent studies indicated a median transplantation free survival of 18 years. More recently, patients with small-duct PSC, who have biochemical and histological features similar to other PSC patients but with a normal cholangiography have been shown to have a better prognosis than classic large-duct PSC. PSC is complicated by cholangiocarcinoma (CCA) which develops in 10-30% of PSC patients depending on the length of follow-up. The diagnosis of an early CCA in the setting of PSC is a major challenge and no consensus on screening strategies exists. No curative therapy for PSC is available at the current time except liver transplantation. The etiopathogenesis of PSC is unknown but the underlying pathophysiology of PSC is beyond the scope of this paper. PMID- 19305377 TI - Management of autoimmune liver disease. AB - The management of autoimmune liver disease can be very challenging. The presentation and natural history of these disorders is highly variable and can be asymptomatic, acute or chronic. Diagnosis requires the skilled interpretation of serological markers and histological changes and sometimes of biliary tract imaging. The major treatment options are immunosuppressive therapy (steroids and azathioprine) in autoimmune hepatitis (AIH) and ursodeoxycholic acid in cholestatic conditions although other treatments continue to be developed and tested. The complications of these diseases, in particular hepatocellular carcinoma and, in PSC, cholangiocarcinoma, remain difficult to monitor or prevent. Liver transplantation remains the only therapeutic option for end stage of liver disease and determining the optimum time for a patient to undergo this procedure requires a sophisticated judgment of the risks and benefits of the procedure as they pertain to an individual patient. PMID- 19305378 TI - The management of hepatorenal syndrome. AB - The hepatorenal syndrome (HRS) is a common complication in advanced liver cirrhosis, and often occurs in patients with ascites and severe circulatory dysfunction. HRS is a functional renal failure which was believed to be the end result of progressive splanchnic vasodilatation. However, recent data have implicated a role of reduced cardiac output as well as endothelial dysfunction in the etiology of HRS. Type 1 HRS is associated with a poor prognosis and often occurs in conjunction with microcirculatory dysfunction in other organs, including the heart, brain and liver. The treatment of type 1 HRS has centered around vasoconstrictors and intravenous hydration, traditionally midodrine and albumin. However, new vasoconstrictors (specifically vasopressin analogues), transjugular intrahepatic portacaval shunts, and albumin dialysis have been introduced as potential therapies. This review will discuss new advances in the diagnosis and pathogenesis of HRS, with an emphasis on the management. PMID- 19305379 TI - Surveillance for violent deaths--national violent death reporting system, 16 States, 2006. AB - PROBLEM/CONDITION: An estimated 50,000 persons die annually in the United States as a result of violence-related injuries. This report summarizes data from CDC's National Violent Death Reporting System (NVDRS) regarding violent deaths from 16 U.S. states for 2006. Results are reported by sex, age group, race/ethnicity, marital status, location of injury, method of injury, circumstances of injury, and other selected characteristics. REPORTING PERIOD COVERED: 2006. DESCRIPTION OF SYSTEM: NVDRS collects data regarding violent deaths obtained from death certificates, coroner/medical examiner reports, and law enforcement reports. NVDRS began operation in 2003 with seven states (Alaska, Maryland, Massachusetts, New Jersey, Oregon, South Carolina, and Virginia) participating; six states (Colorado, Georgia, North Carolina, Oklahoma, Rhode Island, and Wisconsin) joined in 2004 and four (California, Kentucky, New Mexico, and Utah) in 2005, for a total of 17 states. This report includes data from 16 states that collected statewide data; data from California are not included in this report because NVDRS has been implemented only in a limited number of California cities and counties rather than statewide. RESULTS: For 2006, a total of 15,007 fatal incidents involving 15,395 violent deaths occurred in the 16 NVDRS states included in this report. The majority (55.9%) of deaths were suicides, followed by homicides and deaths involving legal intervention (e.g. a suspect is killed by a law enforcement officer in the line of duty)(28.2%), violent deaths of undetermined intent (15.1%), and unintentional firearm deaths (0.7%). Suicides occurred at higher rates among males, American Indians/Alaska Natives (AI/ANs), non-Hispanic whites, and persons aged 45--54 years and occurred most often in a house or apartment and involved the use of firearms. Suicides were precipitated primarily by mental-health, intimate-partner, or physical-health problems or by a crisis during the preceding 2 weeks. Homicides occurred at higher rates among males and persons aged 20--24 years; rates were highest among non-Hispanic black males. The majority of homicides involved the use of a firearm and occurred in a house or apartment or on a street/highway. Homicides were precipitated primarily by arguments and interpersonal conflicts or in conjunction with another crime. Other manners of death and special situations or populations also are highlighted in this report. INTERPRETATION: This report provides a detailed summary of data concerning violent deaths collected by NVDRS for 2006. The results indicate that violent deaths resulting from self-inflicted or interpersonal violence affected adults aged 20--54 years, males, and certain minority populations disproportionately. For many types of violent death, relationship problems, interpersonal conflicts, mental-health problems, and recent crises were among the primary precipitating factors. Because additional information might be reported subsequently as participating states update their findings, the data provided in this report are preliminary. PUBLIC HEALTH ACTION: For the occurrence of violent deaths in the United States to be better understood and ultimately prevented, accurate, timely, and comprehensive surveillance data are necessary. NVDRS data can be used to track the occurrence of violence-related fatal injuries and assist public health authorities in the development, implementation, and evaluation of programs and policies to reduce and prevent violent deaths at the national, state, and local levels. The continued development and expansion of NVDRS is essential to CDC's efforts to reduce the personal, familial, and societal costs of violence. Further efforts are needed to increase the number of states participating in NVDRS, with an ultimate goal of full national representation. PMID- 19305380 TI - Processing the loblolly pine PtGen2 cDNA microarray. AB - PtGen2 is a 26,496 feature cDNA microarray containing amplified loblolly pine ESTs. The array is produced in our laboratory for use by researchers studying gene expression in pine and other conifer species. PtGen2 was developed as a result of our gene discovery efforts in loblolly pine, and is comprised of sequences identified primarily from root tissues, but also from needle and stem. PtGen2 has been tested by hybridizing different Cy-dye labeled conifer target cDNAs, using both amplified and non-amplified indirect labeling methods, and also tested with a number of hybridization and washing conditions. This video focuses on the handling and processing of slides before and after pre-hybridization, as well as after hybridization, using some modifications to procedures developed previously. Also included, in text form only, are the protocols used for the generation, labeling and clean up of target cDNA s, as well as information on software used for downstream data processing. PtGen2 is printed with a proprietary print buffer that contains high concentrations of salt that can be difficult to remove completely. The slides are washed first in a warm SDS solution prior to pre-hybridization. After pre-hybridization, the slides are washed vigorously in several changes of water to complete removal of remaining salts. LifterSlips are then cleaned and positioned on the slides and labeled cDNA is carefully loaded onto the microarray by way of capillary action which provides for even distribution of the sample across the slide, and reduces the chance of bubble incorporation. Hybridization of targets to the array is done at 48 degrees C in high humidity conditions. After hybridization, a series of standard washes are done at 53 degrees C and room temperature for extended times. Processing PtGen2 slides using this technique reduces salt and SDS-derived artifacts often seen when the array is processed less rigorously. Hybridizing targets derived from several different conifer RNA sources, this processing protocol yielded fewer artifacts, reduced background, and provided better consistency among different experimental groups of arrays. PMID- 19305381 TI - P21 Cip1/WAF1 expression is strongly associated with HPV-positive tonsillar carcinoma and a favorable prognosis. AB - Human papillomavirus is involved in the carcinogenesis of tonsillar squamous cell carcinomas. Here, we investigated the expression and the prognostic value of key cell cycle proteins in the pRb and p53 pathways in both human papillomavirus type 16-positive and -negative tonsillar squamous cell carcinomas. Using immunohistochemistry, 77 tonsillar squamous cell carcinomas with known human papillomavirus type 16 status and clinical outcome were analyzed for expression of Ki67, p16(INK4A,) cyclin D1, pRb, p14(ARF), MDM2, p53, p21(Cip1/WAF1), and p27(KIP1). Results were correlated with each other and with clinical and demographic patient data. A total of 35% of tonsillar carcinomas harbored integrated human papillomavirus type 16 DNA and p16(INK4A) overexpression, both being considered essential features for human papillomavirus association. These tumors also showed the overexpression of p14(ARF) (P<0.0001) and p21(Cip1/WAF1) (P=0.001), and downregulation of pRb (P<0.0001) and cyclin D1 (P=0.027) compared with the human papillomavirus-negative cases. Univariate Cox regression analyses revealed a favorable survival rate for non-smokers (P=0.006), as well as for patients with T1-2 tumors (P<0.0001) or tumors showing low expression of cyclin D1 (P=0.028), presence of human papillomavirus and overexpression of p16(INK4A) (P=0.01), p14(ARF) (P=0.02) or p21(Cip1/WAF1) (P=0.004). In multivariate regression analyses, smoking and tumor size, as well as expression of cyclin D1 and p21(Cip1/WAF1), were found to be independent prognostic markers. We conclude that human papillomavirus positivity in tonsillar squamous cell carcinomas strongly correlates with p21(Cip1/WAF1) and p14(ARF) overexpression and downregulation of pRb and cyclin D1. In particular p21(Cip1/WAF1) overexpression is an excellent favorable prognosticator in tonsillar squamous cell carcinomas. PMID- 19305382 TI - Survival impact of psammoma body, stromal calcification, and bone formation in papillary thyroid carcinoma. AB - The presence of calcification is the most significant ultrasonographic finding in evaluating thyroid nodules. Calcifications are more frequently detected in papillary thyroid carcinoma than in other thyroid lesions. However, the clinical significance of calcification, including clinical correlations and impact on survival, and the molecular mechanism responsible for calcification in papillary thyroid carcinoma remain uncertain. We performed a retrospective study of patients with primary common-type papillary thyroid carcinoma to determine the clinical correlations of calcification and its impact on survival. Histologically, calcification was classified as either psammoma bodies, stromal calcification, or bone formation. They were identified in 25, 47, and 13% of all 229 cases of papillary thyroid carcinoma, respectively. The presence of psammoma bodies was significantly correlated with gross lymph node metastasis and stage grouping. Both stromal calcification and bone formation were significantly correlated with patient age. In addition, stromal calcification was associated with pT classification and gross lymph node metastasis. Papillary thyroid carcinoma with, compared to that without, psammoma bodies was associated with poorer disease-free survival. We examined the quantitative expression of BMP-1, a metalloproteinase that is reported to be involved in bone and extracellular matrix formations, and found that its expression was significantly higher in tumors with psammoma bodies or with stromal calcification (P=0.0464 and 0.0272, respectively). These results suggest that the presence of psammoma bodies is a useful predictor of outcome for patients suffering from papillary thyroid carcinoma. PMID- 19305383 TI - Dual expression of alpha-tocopherol-associated protein and estrogen receptor in normal/benign human breast luminal cells and the downregulation of alpha tocopherol-associated protein in estrogen-receptor-positive breast carcinomas. AB - The hormonal carcinogenesis of breast cancer involves hormone-driven cell proliferation and genetic alterations, including oncogene activation and suppressor gene inactivation. However, the predominant genes involved in these processes are currently unknown. Our previous studies identified a gene, namely alpha-tocopherol-associated protein, which is preferentially expressed in normal/benign breast and prostate tissue, but its expression is downregulated in breast and prostate carcinomas. To further examine its function in hormone induced carcinogenesis, we examined if there is an association between alpha tocopherol-associated protein and estrogen-receptor expression in normal/benign breast tissue and in human breast carcinomas. We found that alpha-tocopherol associated protein is coexpressed with estrogen receptor in the luminal cells of normal/benign breast tissue in a scattered manner by immunohistochemical staining of consecutive tissue sections of 20 cases, whereas alpha-tocopherol-associated protein expression is downregulated in 46% (45 of 98) of estrogen receptor/progesterone-receptor-positive, so-called luminal type A or B human breast carcinoma. This is similar to the association of alpha-tocopherol associated protein and androgen receptor expression in normal/benign prostate and prostate carcinomas. In contrast,alpha-tocopherol-associated protein expression is mostly negative in basal, Her2 and triple-negative nonbasal subtypes of high grade breast carcinomas. These findings are consistent with alpha-tocopherol associated protein acting as an antiproliferative factor in estrogen-receptor positive luminal cells in normal/benign breast tissue. alpha-Tocopherol associated protein downregulation may have triggered hormonal carcinogenesis in at least some of the breast carcinomas, providing further, albeit indirect evidence to support a role for vitamin E in breast cancer prevention. PMID- 19305384 TI - Expression of TRAIL and the death receptors DR4 and DR5 correlates with progression of degeneration in human intervertebral disks. AB - Intervertebral disks degenerate far earlier than other musculoskeletal tissues and apoptosis has been suggested to have a vital function in promoting the degeneration process that is strongly associated with back pain. However, the molecular mediators of apoptosis in the intervertebral disk are poorly understood. Fas/FasL, TRAIL/DR4, TRAIL/DR5 and TNF-alpha/TNFR1 are ligand/receptor pairs of the tumor necrosis factor/nerve growth factor family, which are able to induce apoptosis by trimerization of the receptor by its corresponding ligand. We investigated which of these molecules are expressed in intervertebral disks and whether their expression correlates to disk degeneration. Intervertebral disks from 28 donors (age 12-70 years) suffering from scoliosis, vertebrae fracture or disk degeneration were scored histologically for degeneration and analyzed for gene expression of FasL/Fas, TRAIL/DR4, TNF-alpha/TNFR1 and caspase 8. Protein expression of FasL and TRAIL was assessed by immunohistology and apoptotic cell death was quantified by poly(ADP-ribose) polymerase (PARP) p85 staining. Isolated disk cells were analyzed by flow cytometry for Fas, FasL, TRAIL, DR4 and DR5 expression. Gene expression of TRAIL (P=0.002) and caspase 8 (P=0.027) significantly correlated with degeneration. TRAIL expression further correlated with cellularity (P=0.04), muccoid matrix changes (P=0.009) and tears and cleft formation (P=0.019). FasL and TRAIL expression was confirmed by immunohistology and PARP cleavage was significantly associated with degeneration (P=0.027). Flow cytometry on isolated disk cells revealed correlations between DR4 and degeneration (P=0.014), DR4/DR5 double-positive cells and degeneration (P=0.019), as well as DR5 and changes in tissue granularity (P=0.03). This is the first study that shows that intervertebral disk cells express TRAIL, DR4 and DR5, which correlate to the degenerative state of the disk. Therefore, disk cells inherit the molecular machinery to induce and undergo cellular apoptosis, and the frequency of cytokine expression suggests that the TRAIL/DR4/DR5 axis is an important molecular mediator of apoptosis induction in disk tissue. PMID- 19305385 TI - Validation of the 4B5 rabbit monoclonal antibody in determining Her2/neu status in breast cancer. AB - HER2 overexpression in breast cancer is associated with worse clinical outcome. To select patients for anti-Her2-based therapy immunohistochemistry is commonly performed as a first step to assess Her2 status. However, interobserver and interlaboratory variability can significantly compromise adequate assessment of Her2 status. In addition, immunohistochemistry does not always result in an unambiguous test result requiring additional testing for Her2 gene amplification. This study aimed to improve the reliability of Her2 immunohistochemistry by using rabbit monoclonal antibody 4B5 as an alternative to mouse monoclonal antibody CB11 routinely used in our laboratory. Therefore, 283 breast adenocarcinomas were included in a tissue microarray. Immunohistochemistry using the 4B5 and CB11 antibodies, and fluorescence and chromogenic in situ hybridization (FISH or CISH) were performed. Immunohistochemistry was scored by two independent investigators. We found that 4B5 staining was more distinct than CB11 staining. For CB11 staining, there were 12% (BV) and 5% (JW) 2+ scores compared with 4% (BV) and 2% (JW) for 4B5. There was a strong trend towards higher interobserver agreement for 4B5 compared with CB11 (4B5: kappa 0.87, 95% CI 0.79-0.96; CB11: kappa 0.77, 95% CI 0.66-0.88). There were no significant differences in sensitivity, specificity and predictive values between CB11 and 4B5. Our results indicate that the 4B5 antibody provides more robust assessment of immunohistochemical Her2/neu status and will reduce the number of gene amplification tests compared with CB11. However, for tumours with a 2+ score additional gene amplification measurement using FISH or CISH remains necessary. PMID- 19305386 TI - The quest for a better resolution of protein-translocation processes. Conference on the Control, Co-ordination and Regulation of Protein Targeting and Translocation. PMID- 19305387 TI - How are neurons wired to form functional and plastic circuits? Meeting on Axon Guidance, Synaptogenesis & Neural Plasticity. PMID- 19305388 TI - From cancer genomes to cancer models: bridging the gaps. AB - Cancer genome projects are now being expanded in an attempt to provide complete landscapes of the mutations that exist in tumours. Although the importance of cataloguing genome variations is well recognized, there are obvious difficulties in bridging the gaps between high-throughput resequencing information and the molecular mechanisms of cancer evolution. Here, we describe the current status of the high-throughput genomic technologies, and the current limitations of the associated computational analysis and experimental validation of cancer genetic variants. We emphasize how the current cancer-evolution models will be influenced by the high-throughput approaches, in particular through efforts devoted to monitoring tumour progression, and how, in turn, the integration of data and models will be translated into mechanistic knowledge and clinical applications. PMID- 19305389 TI - Filamin B: a scaffold for interferon signalling. PMID- 19305390 TI - Revisiting the COP9 signalosome as a transcriptional regulator. AB - The COP9 signalosome (CSN) is a highly conserved protein complex that was originally described as a repressor of light-dependent growth and transcription in Arabidopsis. The most studied CSN function is the regulation of protein degradation, which occurs primarily through the removal of the ubiquitin-like modifier Nedd8 from cullin-based E3 ubiquitin ligases. This activity can regulate transcription-factor stability and, therefore, transcriptional activity. Recent data suggest that the CSN also regulates transcription on the chromatin by mechanisms that are not yet clearly understood. Furthermore, the CSN subunits CSN5 and CSN2 seem to act as transcriptional coactivators and corepressors, respectively. Here, I re-evaluate the mechanisms by which the CSN acts as a transcriptional regulator, and suggest that they could extend beyond the regulation of protein stability. PMID- 19305391 TI - RANBP2 is an allosteric activator of the conventional kinesin-1 motor protein, KIF5B, in a minimal cell-free system. AB - The association of cargoes to kinesins is thought to promote kinesin activation, yet the validation of such a model with native cargoes is lacking because none is known to activate kinesins directly in an in vitro system of purified components. The RAN-binding protein 2 (RANBP2), through its kinesin-binding domain (KBD), associates in vivo with kinesin-1, KIF5B/KIF5C. Here, we show that KBD and its flanking domains, RAN GTPase-binding domains 2 and 3 (RBD2/RBD3), activate the ATPase activity of KIF5B approximately 30-fold in the presence of microtubules and ATP. The activation kinetics of KIF5B by RANBP2 is biphasic and highly cooperative. Deletion of one of its RBDs lowers the activation of KIF5B threefold and abolishes cooperativity. Remarkably, RBD2-KBD-RBD3 induces unfolding and modest activation of KIF5B in the absence of microtubules. Hence, RANBP2 is the first native and positive allosteric activator known to jump-start and boost directly the activity of a kinesin. PMID- 19305392 TI - A massive hypergiant star as the progenitor of the supernova SN 2005gl. AB - Our understanding of the evolution of massive stars before their final explosions as supernovae is incomplete, from both an observational and a theoretical standpoint. A key missing piece in the supernova puzzle is the difficulty of identifying and studying progenitor stars. In only a single case-that of supernova SN 1987A in the Large Magellanic Cloud-has a star been detected at the supernova location before the explosion, and been subsequently shown to have vanished after the supernova event. The progenitor of SN 1987A was a blue supergiant, which required a rethink of stellar evolution models. The progenitor of supernova SN 2005gl was proposed to be an extremely luminous object, but the association was not robustly established (it was not even clear that the putative progenitor was a single luminous star). Here we report that the previously proposed object was indeed the progenitor star of SN 2005gl. This very massive star was likely a luminous blue variable that standard stellar evolution predicts should not have exploded in that state. PMID- 19305393 TI - Fused has evolved divergent roles in vertebrate Hedgehog signalling and motile ciliogenesis. AB - Hedgehog (Hh) signalling is essential for several aspects of embryogenesis. In Drosophila, Hh transduction is mediated by a cytoplasmic signalling complex that includes the putative serine-threonine kinase Fused (Fu) and the kinesin Costal 2 (Cos2, also known as Cos), yet Fu does not have a conserved role in Hh signalling in mammals. Mouse Fu (also known as Stk36) mutants are viable and seem to respond normally to Hh signalling. Here we show that mouse Fu is essential for construction of the central pair apparatus of motile, 9+2 cilia and offers a new model of human primary ciliary dyskinesia. We found that mouse Fu physically interacts with Kif27, a mammalian Cos2 orthologue, and linked Fu to known structural components of the central pair apparatus, providing evidence for the first regulatory component involved in central pair construction. We also demonstrated that zebrafish Fu is required both for Hh signalling and cilia biogenesis in Kupffer's vesicle. Mouse Fu rescued both Hh-dependent and independent defects in zebrafish. Our results delineate a new pathway for central pair apparatus assembly, identify common regulators of Hh signalling and motile ciliogenesis, and provide insights into the evolution of the Hh cascade. PMID- 19305394 TI - Autophagy enhances the presentation of endogenous viral antigens on MHC class I molecules during HSV-1 infection. AB - Viral proteins are usually processed by the 'classical' major histocompatibility complex (MHC) class I presentation pathway. Here we showed that although macrophages infected with herpes simplex virus type 1 (HSV-1) initially stimulated CD8(+) T cells by this pathway, a second pathway involving a vacuolar compartment was triggered later during infection. Morphological and functional analyses indicated that distinct forms of autophagy facilitated the presentation of HSV-1 antigens on MHC class I molecules. One form of autophagy involved a previously unknown type of autophagosome that originated from the nuclear envelope. Whereas interferon-gamma stimulated classical MHC class I presentation, fever-like hyperthermia and the pyrogenic cytokine interleukin 1beta activated autophagy and the vacuolar processing of viral peptides. Viral peptides in autophagosomes were further processed by the proteasome, which suggests a complex interaction between the vacuolar and MHC class I presentation pathways. PMID- 19305395 TI - Memory T cells in nonlymphoid tissue that provide enhanced local immunity during infection with herpes simplex virus. AB - Effective immunity is dependent on long-surviving memory T cells. Various memory subsets make distinct contributions to immune protection, especially in peripheral infection. It has been suggested that T cells in nonlymphoid tissues are important during local infection, although their relationship with populations in the circulation remains poorly defined. Here we describe a unique memory T cell subset present after acute infection with herpes simplex virus that remained resident in the skin and in latently infected sensory ganglia. These T cells were in disequilibrium with the circulating lymphocyte pool and controlled new infection with this virus. Thus, these cells represent an example of tissue resident memory T cells that can provide protective immunity at points of pathogen entry. PMID- 19305396 TI - C-C chemokine receptor 6-regulated entry of TH-17 cells into the CNS through the choroid plexus is required for the initiation of EAE. AB - Interleukin 17-producing T helper cells (T(H)-17 cells) are important in experimental autoimmune encephalomyelitis, but their route of entry into the central nervous system (CNS) and their contribution relative to that of other effector T cells remain to be determined. Here we found that mice lacking CCR6, a chemokine receptor characteristic of T(H)-17 cells, developed T(H)-17 responses but were highly resistant to the induction of experimental autoimmune encephalomyelitis. Disease susceptibility was reconstituted by transfer of wild type T cells that entered into the CNS before disease onset and triggered massive CCR6-independent recruitment of effector T cells across activated parenchymal vessels. The CCR6 ligand CCL20 was constitutively expressed in epithelial cells of choroid plexus in mice and humans. Our results identify distinct molecular requirements and ports of lymphocyte entry into uninflamed versus inflamed CNS and suggest that the CCR6-CCL20 axis in the choroid plexus controls immune surveillance of the CNS. PMID- 19305397 TI - Molecular docking and ligand specificity in fragment-based inhibitor discovery. AB - Fragment screens have successfully identified new scaffolds in drug discovery, often with relatively high hit rates (5%) using small screening libraries (1,000 10,000 compounds). This raises two questions: would other noteworthy chemotypes be found were one to screen all commercially available fragments (>300,000), and does the success rate imply low specificity of fragments? We used molecular docking to screen large libraries of fragments against CTX-M beta-lactamase. We identified ten millimolar-range inhibitors from the 69 compounds tested. The docking poses corresponded closely to the crystallographic structures subsequently determined. Notably, these initial low-affinity hits showed little specificity between CTX-M and an unrelated beta-lactamase, AmpC, which is unusual among beta-lactamase inhibitors. This is consistent with the idea that the high hit rates among fragments correlate to a low initial specificity. As the inhibitors were progressed, both specificity and affinity rose together, yielding to our knowledge the first micromolar-range noncovalent inhibitors against a class A beta-lactamase. PMID- 19305398 TI - High-energy cathode material for long-life and safe lithium batteries. AB - Layered lithium nickel-rich oxides, Li[Ni(1-x)M(x)]O(2) (M=metal), have attracted significant interest as the cathode material for rechargeable lithium batteries owing to their high capacity, excellent rate capability and low cost. However, their low thermal-abuse tolerance and poor cycle life, especially at elevated temperature, prohibit their use in practical batteries. Here, we report on a concentration-gradient cathode material for rechargeable lithium batteries based on a layered lithium nickel cobalt manganese oxide. In this material, each particle has a central bulk that is rich in Ni and a Mn-rich outer layer with decreasing Ni concentration and increasing Mn and Co concentrations as the surface is approached. The former provides high capacity, whereas the latter improves the thermal stability. A half cell using our concentration-gradient cathode material achieved a high capacity of 209 mA h g(-1) and retained 96% of this capacity after 50 charge-discharge cycles under an aggressive test profile (55 degrees C between 3.0 and 4.4 V). Our concentration-gradient material also showed superior performance in thermal-abuse tests compared with the bulk composition Li[Ni(0.8)Co(0.1)Mn(0.1)]O(2) used as reference. These results suggest that our cathode material could enable production of batteries that meet the demanding performance and safety requirements of plug-in hybrid electric vehicles. PMID- 19305399 TI - Anisotropic self-assembly of spherical polymer-grafted nanoparticles. AB - It is easy to understand the self-assembly of particles with anisotropic shapes or interactions (for example, cobalt nanoparticles or proteins) into highly extended structures. However, there is no experimentally established strategy for creating a range of anisotropic structures from common spherical nanoparticles. We demonstrate that spherical nanoparticles uniformly grafted with macromolecules ('nanoparticle amphiphiles') robustly self-assemble into a variety of anisotropic superstructures when they are dispersed in the corresponding homopolymer matrix. Theory and simulations suggest that this self-assembly reflects a balance between the energy gain when particle cores approach and the entropy of distorting the grafted polymers. The effectively directional nature of the particle interactions is thus a many-body emergent property. Our experiments demonstrate that this approach to nanoparticle self-assembly enables considerable control for the creation of polymer nanocomposites with enhanced mechanical properties. Grafted nanoparticles are thus versatile building blocks for creating tunable and functional particle superstructures with significant practical applications. PMID- 19305400 TI - Phosphodiesterase 1C is dispensable for rapid response termination of olfactory sensory neurons. AB - In the nose, odorants are detected on the cilia of olfactory sensory neurons (OSNs), where a cAMP-mediated signaling pathway transforms odor stimulation into electrical responses. Phosphodiesterase (PDE) activity in OSN cilia has long been thought to account for rapid response termination by degrading odor-induced cAMP. Two PDEs with distinct cellular localization have been found in OSNs: PDE1C in the cilia and PDE4A throughout the cell but absent from the cilia. We disrupted both of these genes in mice and carried out electro-olfactogram analysis. Unexpectedly, eliminating PDE1C did not prolong response termination. Prolonged termination occurred only in mice that lacked both PDEs, suggesting that cAMP degradation by PDE1C in cilia is not a rate-limiting factor for response termination in wild-type mice. Pde1c(-/-) OSNs instead showed reduced sensitivity and attenuated adaptation to repeated stimulation, suggesting that PDE1C may be involved in regulating sensitivity and adaptation. Our observations provide new perspectives on the regulation of olfactory transduction. PMID- 19305401 TI - Transformation of odor representations in target areas of the olfactory bulb. AB - The brain generates coherent perceptions of objects from elementary sensory inputs. To examine how higher-order representations of smells arise from the activation of discrete combinations of glomeruli, we analyzed transformations of activity patterns between the zebrafish olfactory bulb and two of its telencephalic targets, Vv and Dp. Vv is subpallial whereas Dp is the homolog of olfactory cortex. Both areas lack an obvious topographic organization but perform complementary computations. Responses to different odors and their mixtures indicate that Vv neurons pool convergent inputs, resulting in broadened tuning curves and overlapping odor representations. Neuronal circuits in Dp, in contrast, produce a mixture of excitatory and inhibitory synaptic inputs to each neuron that controls action potential firing in an odor-dependent manner. This mechanism can extract information about combinations of molecular features from ensembles of active and inactive mitral cells, suggesting that pattern processing in Dp establishes representations of odor objects. PMID- 19305402 TI - Gating multiple signals through detailed balance of excitation and inhibition in spiking networks. AB - Recent theoretical work has provided a basic understanding of signal propagation in networks of spiking neurons, but mechanisms for gating and controlling these signals have not been investigated previously. Here we introduce an idea for the gating of multiple signals in cortical networks that combines principles of signal propagation with aspects of balanced networks. Specifically, we studied networks in which incoming excitatory signals are normally cancelled by locally evoked inhibition, leaving the targeted layer unresponsive. Transmission can be gated 'on' by modulating excitatory and inhibitory gains to upset this detailed balance. We illustrate gating through detailed balance in large networks of integrate-and-fire neurons. We show successful gating of multiple signals and study failure modes that produce effects reminiscent of clinically observed pathologies. Provided that the individual signals are detectable, detailed balance has a large capacity for gating multiple signals. PMID- 19305403 TI - A sequence element that tunes Escherichia coli tRNA(Ala)(GGC) to ensure accurate decoding. AB - Mutating the rare A32-U38 nucleotide pair at the top of the anticodon loop of Escherichia coli tRNA(Ala)(GGC) to a more common U32-A38 pair results in a tRNA that performs almost normally on cognate codons but is unusually efficient in reading near-cognate codons. Pre-steady state kinetic measurements on E. coli ribosomes show that, unlike the wild-type tRNA(Ala)(GGC), the misreading mutant tRNA(Ala)(GGC) shows rapid GTP hydrolysis and no detectable proofreading on near cognate codons. Similarly, tRNA(Ala)(GGC) mutated to contain C32-G38, a pair that is found in some bacterial tRNA(Ala)(GGC) sequences, was able to decode only the cognate codons, whereas tRNA(Ala)(GGC) containing a more common C32-A38 pair was able to decode all cognate and near-cognate codons tested. We propose that many of the phylogenetically conserved sequence elements present in each tRNA have evolved to suppress translation of near-cognate codons. PMID- 19305404 TI - Bases in the anticodon loop of tRNA(Ala)(GGC) prevent misreading. AB - The bases at positions 32 and 38 in the tRNA anticodon loop are known to have a specific conservation depending upon the anticodon triplets. Here we report that evolutionarily conserved pairs of bases at positions 32 and 38 in tRNA(Ala)(GGC) prevent misreading of a near-cognate valine codon, GUC. The tRNA(Ala)(GGC) molecules with the conserved A32-U38 and C32-G38 pairs do not read GUC, whereas those with three representative nonconserved pairs, U32-U38, U32-A38 and C32-A38, direct the misincorporation of alanine at this valine codon into the peptide chain. Overexpression of the nonconserved tRNA(Ala)(GGC) in Escherichia coli is toxic and prevents cell growth. These results suggested that the bases at positions 32 and 38 in tRNA(Ala)(GGC) evolved to preserve the fidelity of the cognate codon reading. PMID- 19305405 TI - Bacterial frataxin CyaY is the gatekeeper of iron-sulfur cluster formation catalyzed by IscS. AB - Frataxin is an essential mitochondrial protein whose reduced expression causes Friedreich's ataxia (FRDA), a lethal neurodegenerative disease. It is believed that frataxin is an iron chaperone that participates in iron metabolism. We have tested this hypothesis using the bacterial frataxin ortholog, CyaY, and different biochemical and biophysical techniques. We observe that CyaY participates in iron sulfur (Fe-S) cluster assembly as an iron-dependent inhibitor of cluster formation, through binding to the desulfurase IscS. The interaction with IscS involves the iron binding surface of CyaY, which is conserved throughout the frataxin family. We propose that frataxins are iron sensors that act as regulators of Fe-S cluster formation to fine-tune the quantity of Fe-S cluster formed to the concentration of the available acceptors. Our observations provide new perspectives for understanding FRDA and a mechanistic model that rationalizes the available knowledge on frataxin. PMID- 19305406 TI - SUnSET, a nonradioactive method to monitor protein synthesis. AB - We developed a nonradioactive fluorescence-activated cell sorting-based assay, called surface sensing of translation (SUnSET), which allows the monitoring and quantification of global protein synthesis in individual mammalian cells and in heterogeneous cell populations. We demonstrate here, using mouse dendritic and T cells as a model, that SUnSET offers a technical alternative to classical radioactive labeling methods for the study of mRNA translation and cellular activation. PMID- 19305407 TI - Global mapping of protein-DNA interactions in vivo by digital genomic footprinting. AB - The orchestrated binding of transcriptional activators and repressors to specific DNA sequences in the context of chromatin defines the regulatory program of eukaryotic genomes. We developed a digital approach to assay regulatory protein occupancy on genomic DNA in vivo by dense mapping of individual DNase I cleavages from intact nuclei using massively parallel DNA sequencing. Analysis of >23 million cleavages across the Saccharomyces cerevisiae genome revealed thousands of protected regulatory protein footprints, enabling de novo derivation of factor binding motifs and the identification of hundreds of new binding sites for major regulators. We observed striking correspondence between single-nucleotide resolution DNase I cleavage patterns and protein-DNA interactions determined by crystallography. The data also yielded a detailed view of larger chromatin features including positioned nucleosomes flanking factor binding regions. Digital genomic footprinting should be a powerful approach to delineate the cis regulatory framework of any organism with an available genome sequence. PMID- 19305408 TI - Common variants at ten loci influence QT interval duration in the QTGEN Study. AB - QT interval duration, reflecting myocardial repolarization on the electrocardiogram, is a heritable risk factor for sudden cardiac death and drug induced arrhythmias. We conducted a meta-analysis of three genome-wide association studies in 13,685 individuals of European ancestry from the Framingham Heart Study, the Rotterdam Study and the Cardiovascular Health Study, as part of the QTGEN consortium. We observed associations at P < 5 x 10(-8) with variants in NOS1AP, KCNQ1, KCNE1, KCNH2 and SCN5A, known to be involved in myocardial repolarization and mendelian long-QT syndromes. Associations were found at five newly identified loci, including 16q21 near NDRG4 and GINS3, 6q22 near PLN, 1p36 near RNF207, 16p13 near LITAF and 17q12 near LIG3 and RFFL. Collectively, the 14 independent variants at these 10 loci explain 5.4-6.5% of the variation in QT interval. These results, together with an accompanying paper, offer insights into myocardial repolarization and suggest candidate genes that could predispose to sudden cardiac death and drug-induced arrhythmias. PMID- 19305409 TI - Common variants at ten loci modulate the QT interval duration in the QTSCD Study. AB - The QT interval, a measure of cardiac repolarization, predisposes to ventricular arrhythmias and sudden cardiac death (SCD) when prolonged or shortened. A common variant in NOS1AP is known to influence repolarization. We analyze genome-wide data from five population-based cohorts (ARIC, KORA, SardiNIA, GenNOVA and HNR) with a total of 15,842 individuals of European ancestry, to confirm the NOS1AP association and identify nine additional loci at P < 5 x 10(-8). Four loci map near the monogenic long-QT syndrome genes KCNQ1, KCNH2, SCN5A and KCNJ2. Two other loci include ATP1B1 and PLN, genes with established electrophysiological function, whereas three map to RNF207, near LITAF and within NDRG4-GINS3-SETD6 CNOT1, respectively, all of which have not previously been implicated in cardiac electrophysiology. These results, together with an accompanying paper from the QTGEN consortium, identify new candidate genes for ventricular arrhythmias and SCD. PMID- 19305410 TI - Natural variation at the DEP1 locus enhances grain yield in rice. AB - Grain yield is controlled by quantitative trait loci (QTLs) derived from natural variations in many crop plants. Here we report the molecular characterization of a major rice grain yield QTL that acts through the determination of panicle architecture. The dominant allele at the DEP1 locus is a gain-of-function mutation causing truncation of a phosphatidylethanolamine-binding protein-like domain protein. The effect of this allele is to enhance meristematic activity, resulting in a reduced length of the inflorescence internode, an increased number of grains per panicle and a consequent increase in grain yield. This allele is common to many Chinese high-yielding rice varieties and likely represents a relatively recent introduction into the cultivated rice gene pool. We also show that a functionally equivalent allele is present in the temperate cereals and seems to have arisen before the divergence of the wheat and barley lineages. PMID- 19305411 TI - Many X-linked microRNAs escape meiotic sex chromosome inactivation. AB - Meiotic sex chromosome inactivation (MSCI) during spermatogenesis is characterized by transcriptional silencing of genes on both the X and Y chromosomes in mid-to-late pachytene spermatocytes. MSCI is believed to result from meiotic silencing of unpaired DNA because the X and Y chromosomes remain largely unpaired throughout first meiotic prophase. However, unlike X-chromosome inactivation in female embryonic cells, where 25-30% of X-linked structural genes have been reported to escape inactivation, previous microarray- and RT-PCR-based studies of expression of >364 X-linked mRNA-encoding genes during spermatogenesis have failed to reveal any X-linked gene that escapes the silencing effects of MSCI in primary spermatocytes. Here we show that many X-linked miRNAs are transcribed and processed in pachytene spermatocytes. This unprecedented escape from MSCI by these X-linked miRNAs suggests that they may participate in a critical function at this stage of spermatogenesis, including the possibility that they contribute to the process of MSCI itself, or that they may be essential for post-transcriptional regulation of autosomal mRNAs during the late meiotic and early postmeiotic stages of spermatogenesis. PMID- 19305412 TI - Heterotypic interactions enabled by polarized neutrophil microdomains mediate thromboinflammatory injury. AB - Selectins and their ligands mediate leukocyte rolling, allowing interactions with chemokines that lead to integrin activation and arrest. Here we show that E selectin is crucial for generating a secondary wave of activating signals, transduced specifically by E-selectin ligand-1, that induces polarized, activated alpha(M)beta(2) integrin clusters at the leading edge of crawling neutrophils, allowing capture of circulating erythrocytes or platelets. In a humanized mouse model of sickle cell disease, the capture of erythrocytes by alpha(M)beta(2) microdomains leads to acute lethal vascular occlusions. In a model of transfusion related acute lung injury, polarized neutrophils capture circulating platelets, resulting in the generation of oxidative species that produce vascular damage and lung injury. Inactivation of E-selectin or alpha(M)beta(2) prevents tissue injury in both inflammatory models, suggesting broad implications of this paradigm in thromboinflammatory diseases. These results indicate that endothelial selectins can influence neutrophil behavior beyond its canonical rolling step through delayed, organ-damaging, polarized activation. PMID- 19305413 TI - Stimulation of tumor growth and angiogenesis by low concentrations of RGD-mimetic integrin inhibitors. AB - Inhibitors of alpha(v)beta(3) and alpha(v)beta(5) integrin have entered clinical trials as antiangiogenic agents for cancer treatment but generally have been unsuccessful. Here we present in vivo evidence that low (nanomolar) concentrations of RGD-mimetic alpha(v)beta(3) and alpha(v)beta(5) inhibitors can paradoxically stimulate tumor growth and tumor angiogenesis. We show that low concentrations of these inhibitors promote VEGF-mediated angiogenesis by altering alpha(v)beta(3) integrin and vascular endothelial growth factor receptor-2 trafficking, thereby promoting endothelial cell migration to VEGF. The proangiogenic effects of low concentrations of RGD-mimetic integrin inhibitors could compromise their efficacy as anticancer agents and have major implications for the use of RGD-mimetic compounds in humans. PMID- 19305414 TI - Bioluminescence imaging of myeloperoxidase activity in vivo. AB - The myeloperoxidase (MPO) system of activated phagocytes is central to normal host defense mechanisms, and dysregulated MPO contributes to the pathogenesis of inflammatory disease states ranging from atherosclerosis to cancer. Here we show that upon systemic administration, the small molecule luminol enables noninvasive bioluminescence imaging (BLI) of MPO activity in vivo. Luminol-BLI allowed quantitative longitudinal monitoring of MPO activity in animal models of acute dermatitis, mixed allergic contact hypersensitivity, focal arthritis and spontaneous large granular lymphocytic tumors. Bioluminescence colocalized with histological sites of inflammation and was totally abolished in gene-deleted Mpo( /-) mice, despite massive tissue infiltration of neutrophils and activated eosinophils, indicating that eosinophil peroxidase did not contribute to luminol BLI in vivo. Thus, luminol-BLI provides a noninvasive, specific and highly sensitive optical readout of phagocyte-mediated MPO activity in vivo and may enable new diagnostic applications in a wide range of acute and chronic inflammatory conditions. PMID- 19305415 TI - Delivering proteins for export from the cytosol. AB - Correct protein function depends on delivery to the appropriate cellular or subcellular compartment. Following the initiation of protein synthesis in the cytosol, many bacterial and eukaryotic proteins must be integrated into or transported across a membrane to reach their site of function. Whereas in the post-translational delivery pathway ATP-dependent factors bind to completed polypeptides and chaperone them until membrane translocation is initiated, a GTP dependent co-translational pathway operates to couple ongoing protein synthesis to membrane transport. These distinct pathways provide different solutions for the maintenance of proteins in a state that is competent for membrane translocation and their delivery for export from the cytosol. PMID- 19305416 TI - Polo-like kinases: conservation and divergence in their functions and regulation. AB - Polo-like kinases (Plks) are potent regulators of M phase that are conserved from yeasts to humans. Their roles in mitotic entry, spindle pole functions and cytokinesis are broadly conserved despite physical and molecular differences in these processes in disparate organisms. Plks are characterized by their Polo-box domain, which mediates protein interactions. They are additionally controlled by phosphorylation, proteolysis and transcription, depending on the biological context. Plks are now recognized to link cell division to developmental processes and to function in differentiated cells. A comparison of Plk function and regulation between organisms offers insight into the rich variations of cell division. PMID- 19305417 TI - Beta-catenin hits chromatin: regulation of Wnt target gene activation. AB - The canonical Wnt pathway has gathered much attention in recent years owing to its fundamental contribution to metazoan development, tissue homeostasis and human malignancies. Wnt target gene transcription is regulated by nuclear beta catenin, and genetic assays have revealed various collaborating protein cofactors. Their daunting number and diverse nature, however, make it difficult to arrange an orderly picture of the nuclear Wnt transduction events. Yet, these findings emphasize that beta-catenin-mediated transcription affects chromatin. How does beta-catenin cope with chromatin regulation to turn on Wnt target genes? PMID- 19305418 TI - Is the high virulence of HIV-1 an unfortunate coincidence of primate lentiviral evolution? AB - In the subset of primate lentiviruses that contain a vpu gene - HIV-1 and its simian precursors - the Nef protein has lost the ability to down-modulate CD3, block T cell activation and suppress programmed death. Vpu counteracts a host restriction factor induced by the inflammatory cytokine interferon-alpha. I propose that the acquisition of vpu may have allowed the viral lineage that gave rise to HIV-1 to evolve towards greater pathogenicity by removing the selective pressure for a protective Nef function that prevents damagingly high levels of immune activation. PMID- 19305419 TI - Association of serum sodium concentration with coronary atherosclerosis in China: follow-up study. AB - AIM: The aim of this study was to test the hypothesis that lower serum sodium may be associated with increased cardiovascular events and all-cause mortality by means of long-term follow-up of subjects with coronary atherosclerosis in a prospective, hospital-based epidemiological study in China. METHODS: A prospective, hospital-based epidemiological design was used. The study population consisted of 1069 consecutive patients who were scheduled to undergo coronary angiography for suspected or known coronary atherosclerosis. The severity of coronary atherosclerosis was defined using Gensini's score system. Age, sex adjusted hazard ratios (HR) and 95% confidence intervals (CI) for the quartiles of serum sodium concentration were estimated with Cox proportional hazard models, using quartile 1 as the reference. Cox proportional hazard models were also constructed to estimate the hazard ratios and 95% confidence intervals for all cause mortality and final end-point events by serum sodium quartile and to adjust for potentially confounding variables. Multivariate models were adjusted for the following variables: age, sex, smoking status, alcohol consumption, body mass index, blood pressure, potassium, chloride, total cholesterol, triglycerides, fasting blood glucose, urea, creatinine, uric acid, and Gensini's score. RESULTS: During the median 2.86 years (3011.66 person-years) of follow-up, 176 final end point events were documented. These events included 79 deaths and 97 readmissions for coronary heart disease. There was a statistically significant inverse association of serum sodium with all-cause mortality (P<0.001). After full adjustment comparing the highest serum sodium quartile to the lowest, there was a non-significant inverse association with all-cause mortality, with an adjusted hazard ratio (95% CI) of 0.67 (0.25-1.80). After adjustment for age and sex, the hazard ratio and 95% CI for final end-point events across increasing quartiles of serum sodium concentration were 1.00, 0.85 (0.59-1.22), 0.52 (0.34-0.82), and 0.31 (0.19-0.49). After full adjustment comparing the highest serum sodium quartile to the lowest, there was a statistically significant inverse association with final end-point events, with an adjusted hazard ratio (95% CI) of 0.46 (0.26 0.81). CONCLUSION: The serum sodium concentration showed a statistically significant negative association with coronary events and all-cause mortality in subjects with coronary atherosclerosis; the actual mechanism underlying this association needs further study. PMID- 19305420 TI - Composite glycidyl methacrylated dextran (Dex-GMA)/gelatin nanoparticles for localized protein delivery. AB - AIM: Localized delivery of growth factors has significant potential as a future therapeutic strategy in tissue engineering and regenerative medicine. A nanoparticle vehicle was created and evaluated in this study with the intent to deliver growth factors for periodontal regeneration. METHODS: Novel composite nanoparticles based on glycidyl methacrylate derivatized dextrans (Dex-GMA) and gelatin were fabricated by a facile method without using any organic solvents. The configurations of the resultant nanoparticles were evaluated by transmission electron microscopy, scanning electron microscopy, and atomic force microscope. Their surfaces were characterized by zeta-potential measurements, after which their properties including swelling, degradation, drug release, and cytotoxicity were also investigated using in vitro models. RESULTS: The particle size of Dex GMA/gelatin nanoparticles (DG-NPs) ranged from 20 to 100 nm and showed a mono disperse size distribution (mean diameter 53.7 nm) and a strongly negative surface zeta potential (-20 mV). The DG-NPs were characterized by good swelling and degradation properties in media including dextranase. The in vitro drug release studies showed that the efficient bone morphogenetic protein (BMP) release from DG-NPs was maintained for more than 12 d under degradation conditions, where more than 90% of the loaded BMP was released. No any relevant cell damage caused by DG-NPs was found in the cytotoxicity tests for a period of 24 h. CONCLUSION: These combined results demonstrate that DG-NPs fulfill the basic prerequisites for growth factor delivery. With further in vivo studies, those nanoparticles may offer a promising vehicle for the delivery of active drugs to the periodontium. PMID- 19305421 TI - Isoform-specific regulation of the Na+ -K+ pump by adenosine in guinea pig ventricular myocytes. AB - AIM: The present study investigated the effect of adenosine on Na(+)-K(+) pumps in acutely isolated guinea pig (Cavia sp.) ventricular myocytes. METHODS: The whole-cell, patch-clamp technique was used to record the Na(+)-K(+) pump current (I(p)) in acutely isolated guinea pig ventricular myocytes. RESULTS: Adenosine inhibited the high DHO-affinity pump current (I(h)) in a concentration-dependent manner, which was blocked by the selective adenosine A(1) receptor antagonist DPCPX and the general protein kinase C (PKC) antagonists staurosporine, GF 109203X or the specific delta isoform antagonist rottlerin. In addition, the inhibitory action of adenosine was mimicked by a selective A(1) receptor agonist CCPA and a specific activator peptide of PKC-delta, PP114. In contrast, the selective A(2A) receptor agonist CGS21680 and A(3) receptor agonist Cl-IB-MECA did not affect I(h). Application of the selective A(2A) receptor antagonist SCH58261 and A(3) receptor antagonist MRS1191 also failed to block the effect of adenosine. Furthermore, H89, a selective protein kinase A (PKA) antagonist, did not exert any effect on adenosine-induced I(h) inhibition. CONCLUSION: The present study provides the electrophysiological evidence that adenosine can induce significant inhibition of I(h) via adenosine A(1) receptors and the PKC delta isoform. PMID- 19305422 TI - Sinomenine influences capacity for invasion and migration in activated human monocytic THP-1 cells by inhibiting the expression of MMP-2, MMP-9, and CD147. AB - AIM: The aim of this study was to investigate the mechanism of the effects of Sinomenine (SIN) on the invasion and migration ability of activated human monocytic THP-1 cells (A-THP-1). Sinomenine is a pure alkaloid extracted from the Chinese medical plant Sinomenium acutum. METHODS: Human monocytic THP-1 cells were induced to differentiate into macrophages with phorbol 12-myristate 13 acetate (PMA). Cells were treated with different concentrations of SIN. The invasion and migration ability of cells was tested by in vitro transwell assays. The levels of CD147 and MMPs were evaluated by flow cytometric analysis and zymographic analysis, respectively. The mRNA expression of CD147, MMP-2, and MMP 9 was measured by RT-PCR. RESULTS: The invasion and migration ability of A-THP-1 cells was significantly inhibited by SIN in a concentration-dependent fashion; at the same time, the levels of CD147, MMP-2, and MMP-9 were markedly down regulated. This inhibitory effect was most notable at concentrations of 0.25 mmol/L and 1.00 mmol/L (P<0.01). CONCLUSION: A possible mechanism of the inhibitory effect of SIN on cell invasion and migration ability is repression of the expression of MMP-2 and MMP-9, which strongly correlates with the inhibition of CD147 activity. PMID- 19305423 TI - Induction of G2/M arrest by pseudolaric acid B is mediated by activation of the ATM signaling pathway. AB - AIM: The aim of this study was to investigate the mechanism of pseudolaric acid B (PLAB)-induced cell cycle arrest in human melanoma SK-28 cells. METHODS: Cell growth inhibition was detected by MTT assay, the cell cycle was analyzed by flow cytometry, and protein expression was examined by Western blot analysis. RESULTS: PLAB inhibited the growth of human melanoma cells and induced G(2)/M arrest in SK 28 cells, accompanied by an up-regulation of Cdc2 phosphorylation and a subsequent down-regulation of Cdc2 expression. Furthermore, PLAB decreased the expression of Cdc25C phosphatase and increased the expression of Wee1 kinase. Meanwhile, a reduction in Cdc2 activity was partly due to induction of the expression of p21(waf1/cip1) in a p53-dependent manner. In addition, PLAB activated the checkpoint kinase, Chk2, and increased the expression of p53, two major targets of ATM kinase. These effects were inhibited by caffeine, an ATM kinase inhibitor. We also found that PLAB significantly enhanced ATM kinase activity. CONCLUSION: Taken together, these results suggest that PLAB induced G(2)/M arrest in human melanoma cells via a mechanism involving the activation of ATM, and the effect of PLAB on Cdc2 activity was mediated via interactions with the Chk2-Cdc25C and p53 signalling pathways, two distinct downstream pathways of ATM. PLAB may be a promising chemopreventive agent for treating human melanoma. PMID- 19305424 TI - Anti-hypoxic effect of ginsenoside Rbl on neonatal rat cardiomyocytes is mediated through the specific activation of glucose transporter-4 ex vivo. AB - AIM: The aim of this study was to investigate whether Gs-Rbl relieves the CoCl(2) induced apoptosis of hypoxic neonatal rat cardiomyocytes and in which the role of glucose transporter-4 (GLUT-4). METHODS: Gs-Rbl (0, 10, 50, 100, 200, 400, and 500 micromol/L), adenine 9-beta-D-arabinofuranoside (ara A, 500 micromol/L; AMPK inhibitor) and wortmannin (0.5 micromol/L; PI3K inhibitor) only in combination with 200 micromol/L Gs-Rbl were administered in hypoxic cardiomyocytes, which were induced by 500 micromol/L CoCl(2) for 12 h. Then, the apoptotic rate (AR), 2 [(3)H]-deoxy-D-glucose (2-[(3)H]-DG) uptake, and the expression of GLUT-4 (including in plasma membrane, PM), phospho-AMPKalpha (Thr172), AMPKalpha and Akt in cells were assayed. RESULTS: Compared with simple hypoxia (0 micromol/L Gs Rbl), Gs-Rb1 greater than 10 micromol/L significantly decreased the apoptotic rate (P<0.01) and significantly increased 2-[(3)H]-DG uptake (P<0.01), GLUT-4 content in cells and PM (P<0.01), AMPK activity (P<0.01) and Akt (P<0.01) levels in a dose-dependent manner. AMPK activity was completely suppressed by ara-A, just as Akt was suppressed by wortmannin. The AR, glucose uptake and GLUT-4 levels in cells and PM were partly down-regulated by ara-A or wortmannin. CONCLUSION: Gs-Rb1 may protect neonatal rat cardiomyocytes from apoptosis induced by CoCl(2). The anti-apoptotic effect of Gs-Rb1 may occur by improving glucose uptake, in which GLUT-4 translocation and expression played a key role. Both the AMPK and the PI3K/Akt pathways may take part in the anti-hypoxic efficacy of Gs Rb1. PMID- 19305425 TI - Upregulation of PIP3-dependent Rac exchanger 1 (P-Rex1) promotes prostate cancer metastasis. AB - Excessive activation of G-protein-coupled receptor (GPCR) and receptor tyrosine kinase (RTK) pathways has been linked to prostate cancer metastasis. Rac activation by guanine nucleotide exchange factors (GEFs) plays an important role in directional cell migration, a critical step of tumor metastasis cascades. We found that the upregulation of P-Rex1, a Rac-selective GEF synergistically activated by Gbetagamma freed during GPCR signaling, and PIP3, generated during either RTK or GPCR signaling, strongly correlates with metastatic phenotypes in both prostate cancer cell lines and human prostate cancer specimens. Silencing endogenous P-Rex1 in metastatic prostate cancer PC-3 cells selectively inhibited Rac activity and reduced cell migration and invasion in response to ligands of both epidermal growth factor receptor and G-protein-coupled CXC chemokine receptor 4. Conversely, expression of recombinant P-Rex1, but not its 'GEF-dead' mutant, in non-metastatic prostate cancer cells, such as CWR22Rv1, increased cell migration and invasion through Rac-dependent lamellipodia formation. More importantly, using a mouse xenograft model, we showed that the expression of P Rex1, but not its mutant, induced lymph node metastasis of CWR22Rv1 cells without an effect on primary tumor growth. Thus, by functioning as a coincidence detector of chemotactic signals from both GPCRs and RTKs, P-Rex1-dependent activation of Rac promotes prostate cancer metastasis. PMID- 19305426 TI - The von Hippel-Lindau protein sensitizes renal carcinoma cells to apoptotic stimuli through stabilization of BIM(EL). AB - von Hippel-Lindau (VHL) disease is caused by germ-line mutations in the VHL tumor suppressor gene and is the most common cause of inherited renal cell carcinoma (RCC). Mutations in the VHL gene also occur in a large majority of sporadic cases of clear-cell RCC, which have high intrinsic resistance to chemotherapy and radiotherapy. Here we show that VHL-deficient RCC cells express lower levels of the proapoptotic Bcl-2 family protein BIM(EL) and are more resistant to etoposide and UV radiation-induced death compared to the same cells stably expressing the wild-type VHL protein (pVHL). Reintroducing pVHL into VHL-null cells increased the half-life of BIM(EL) protein without affecting its mRNA expression, and overexpressing pVHL inhibited BIM(EL) polyubiquitination. Suppressing pVHL expression with RNA interference resulted in a decrease in BIM(EL) protein and a corresponding decrease in the sensitivity of RCC cells to apoptotic stimuli. Directly inhibiting BIM(EL) expression in pVHL-expressing RCC cells caused a similar decrease in cell death. These results demonstrate that pVHL acts to promote BIM(EL) protein stability in RCC cells, and that destabilization of BIM(EL) in the absence of pVHL contributes to the increased resistance of VHL null RCC cells to certain apoptotic stimuli. PMID- 19305427 TI - Familial breast cancer screening reveals an alteration in the RAP80 UIM domain that impairs DNA damage response function. AB - Germline mutations in two major susceptibility genes, BRCA1 and BRCA2, account for nearly 20% of familial breast cancers. A majority of the remaining genetic factors involved in heritable breast cancer susceptibility are, however, unknown. Recently, a new BRCA1-interacting protein, receptor associated protein 80 (RAP80), was identified. RAP80 plays an important role in BRCA1-mediated DNA damage responses (DDRs) by recruiting BRCA1 to DNA double-strand breaks (DSBs). A comprehensive screening of DNA from affected index cases of 112 BRCA1/BRCA2 mutation-negative Finnish breast cancer families revealed altogether 10 alterations in RAP80, one of which, c.241-243delGAA, resulted in a single glutamic acid deletion at residue 81 in a highly conserved region of ubiquitin interaction motif 1. The resultant delE81 protein product displayed significantly reduced ubiquitin binding and DSB localization. Expression of the RAP80 delE81 allele impaired both BRCA1 and ABRA1 DSB recruitment, thus compromising BRCA1 mediated DDR signaling. Compared with wild-type RAP80, expression of the delE81 allele was associated with a significant increase in cytogenetically detectable chromosomal aberrations, particularly chromatid breaks. Although evidently quite rare, these results suggest that critical constitutional mutations in RAP80 abrogate DDR function and may be involved in genetic predisposition to cancer. PMID- 19305428 TI - The role of cooperativity with Src in oncogenic transformation mediated by non small cell lung cancer-associated EGF receptor mutants. AB - Non-small cell lung cancer (NSCLC)-associated epidermal growth factor receptor (EGFR) mutants are constitutively active and induce ligand-independent transformation in non-malignant cell lines. We investigated the possibility that the ability of mutant EGFRs to transform cells reflects a constitutive cooperativity with Src using a system in which the overexpression of mutant, but not wild-type, EGFR induced anchorage-independent cell growth. Src was constitutively activated and showed enhanced interaction with mutant EGFRs, suggesting that constitutive EGFR-Src cooperativity may contribute to mutant EGFR mediated oncogenesis. Indeed, the mutant EGFR-mediated cell transformation was inhibited by Src- as well as EGFR-directed inhibitors. Importantly, a tyrosine to phenylalanine mutation of the major Src phosphorylation site on EGFR, Y845, reduced the constitutive phosphorylation of NSCLC-EGFR mutants, as well as that of STAT3, Akt, Erk and Src, and reduced the mutant EGFR-Src association as well as proliferation, migration and anchorage-independent growth. Reduced anchorage independent growth and migration were also observed when dominant-negative-Src was expressed in mutant EGFR-expressing cells. Overall, our findings show that mutant EGFR-Src interaction and cooperativity play critical roles in constitutive engagement of the downstream signaling pathways that allow NSCLC-associated EGFR mutants to mediate oncogenesis, and support the rationale to target Src-dependent signaling pathways in mutant EGFR-mediated malignancies. PMID- 19305430 TI - Cardiac and Kidney Weight Indices following Dietary Salt Loading and/or Chronic Nitric Oxide Synthase Inhibition in the Hooded (Aguti) Rat. AB - OBJECTIVES: This study was designed to investigate if dietary salt-loading and/or chronic Nitric Oxide Synthase (NOS) inhibition are associated with cardiac hypertrophy and changes in renal mass in the hooded (Aguti) rat. METHODS: Hooded rats 8-10 weeks old were divided into 4 groups viz: control, salt, L-NAME (N?-L Arginine Methyl Ester) and salt + L-NAME. Control rats were given a normal rat diet and water. Hypertension was induced in hooded rats by giving the following treatments: rats in the salt group were given an 8% NaCl diet and water for 6 weeks; rats in the L-NAME group were fed the normal rat diet and given water containing L-NAME at a dose of 100mg/Kg/day for 4 weeks; rats in the salt + L NAME group were given both treatments. The rats (n = 8 per group) were anaesthetized and the hearts and kidneys excised. The cardiac weight indices and the kidney weights were measured. RESULTS: The cardiac weight, cardiac weight index, left and right ventricular weight indices and kidney weight showed no significant difference in the test groups compared to control. Kidney weight/body weight (g/100g body weight) increased significantly (P<0.05) in salt+L-NAME rats (0.35 +/- 0.015) compared to control (0.31 +/- 0.013), salt-loaded (0.29 +/- 0.013) and L-NAME rats (0.20 +/- 0.010). CONCLUSIONS: Six weeks of dietary salt loading and /or four weeks of L-NAME-loading were not associated with cardiac hypertrophy in the hooded rat. However a combination of both manoeuvres resulted in renal hypertrophy. PMID- 19305429 TI - NAC-1, a potential stem cell pluripotency factor, contributes to paclitaxel resistance in ovarian cancer through inactivating Gadd45 pathway. AB - Nucleus accumbens-1 (Nac1 or NAC-1) belongs to the BTB/POZ (Pox virus and Zinc finger/Bric-a-brac Tramtrack Broad complex) transcription factor family and is a novel protein that potentially participates in self-renewal and pluripotency in embryonic stem cells. In human cancer, NAC-1 is upregulated in several types of neoplasms, but particularly in recurrent chemoresistant ovarian carcinomas, suggesting a biological role for NAC-1 in the development of drug resistance in ovarian cancer. We have assessed this possibility and shown a correlation between NAC-1 expression and ex vivo paclitaxel resistance in ovarian serous carcinoma tissues and cell lines. We found that expression of Gadd45-gamma-interacting protein 1 (Gadd45gip1), a downstream target negatively regulated by NAC-1, was reduced in paclitaxel-resistant cells. Ectopic expression of NAC-1 or knockdown of Gadd45gip1 conferred paclitaxel resistance, whereas NAC-1 knockdown or ectopic expression of Gadd45gip1 increased paclitaxel sensitivity. Furthermore, silencing NAC-1 expression or disrupting NAC-1 homodimerization by a dominant negative NAC 1 protein that contained only the BTB/POZ domain induced the expression of Gadd45gamma, which interacted with Gadd45gip1. Reducing Gadd45gamma expression by small hairpin RNAs partially enhanced paclitaxel resistance. Thus, this study provides new evidence that NAC-1 upregulation and homodimerization contribute to tumor recurrence by equipping ovarian cancer cells with the paclitaxel-resistant phenotype through negative regulation of the Gadd45 pathway. PMID- 19305431 TI - Knowledge and practices of poultry workers on prevention of avian flu in osogbo, osun state, Nigeria. AB - INTRODUCTION: Nigeria recorded its first case of avian flu among birds early in the year 2006 and by the third quarter of the year, about sixteen states of the country had been affected. Finding out the knowledge and practices of persons in close contact with birds as regards the avian flu would help to identify areas in need of focused attention and alert the coordinating agencies of the magnitude and prevalence of practices which may encourage the spread of the disease. METHODOLOGY: This was a descriptive cross-sectional survey of poultry owners and workers in Osogbo, Osun State, southwestern Nigeria. Information was obtained from 65 of the 100 registered members of the poultry association who consented to answering questions on the pre-tested semi-structured questionnaires. RESULT: In this study, 49.1% (28) of the study sample knew of avian flu, and 46.4% (13) of these knew that the disease was present in Nigeria. Only 50% (14) of those who knew about the disease felt it could affect human beings. For those who have heard of the disease before, 85.7% (24) knew it could be transmitted from sick birds to humans. Practices which favour the spread of the virus that were engaged in by the respondents included using bird droppings as manure (71.9%), and feeding fish with intestines of killed chicken (66.7%). CONCLUSION: The study shows a low level of awareness of avian flu among the poultry workers and owners. It is recommended that massive education should be embarked upon for groups occupying strategic positions in disease epidemiology, as well as all stake holders in poultry farming. PMID- 19305432 TI - Enhancing data management skills of primary health care workers in enugu state, Nigeria. AB - OBJECTIVE: The objective of this study was to evaluate the outcome of a training scheme on data collection, recording, reporting and utilisation practices among primary health care workers in Enugu State. METHODS: This is a "before and after" intervention study conducted among 107 Primary Health Care (PHC) workers in Enugu State. A multistage sampling technique was used. A pre-tested, semi-structured questionnaire and an observation checklist were the instruments used in data collection pre and post-intervention. RESULTS: The workers in this study pre intervention recorded data using health management information system (HMIS) forms, registers and exercise books. Seven (77.8%) and 6(66.2%) of the study and control health centres respectively kept monthly health center copies of HMIS form 000. These forms and registers were neither completely and correctly filled nor were kept up-to-date. Post-intervention, the reported and observed practices improved significantly (p<0.001) in the study unlike in the control health centress. Pre-intervention, none of the health centres entered data into the register 1-2weeks prior to this study and none remitted same monthly to the monitoring and evaluation units of the LGAs. Post-intervention, early data entry, reporting and local uses of data improved significantly (p<0.001) in the study group unlike in the control group. CONCLUSION: The data management skills of PHC workers studied were generally inadequate pre-intervention. Training greatly enhanced their overall data management skills. PMID- 19305433 TI - Effect of Demographic Variables on Cytomegalovirus Antibody Seropositivity among Prospective Blood Donors in Jos, Nigeria. AB - BACKGROUND: Cytomegalovirus, otherwise called human herpes virus type 5, is a transfusion transmissible pathogen responsible for significant morbidity and mortality among various groups of individuals with damaged or impaired host immunity. Although it has a worldwide distribution, the infection is thought to be more widespread in developing countries where certain demographic factors and living conditions are thought to be responsible for the observed increased prevalence of this infection. OBJECTIVES: The aim of this study is to evaluate the effect of living conditions, using certain selected demographic parameters, on CMV seropositivity, among blood donors in Jos. PATIENTS AND METHODS: A total of 200 prospective blood donors who presented for bleeding were recruited consecutively into the study. Questionnaire was used to obtain the personal, educational, social and demographic data of donors. Screening for CMV antibodies was performed on each serum sample using ELISA kit ("DIALAB", Austria) (www.dialab.at). RESULTS: The relationship between the extent of crowding and CMV status of donors was statistically significant, p<0.05. Other studied donor demographic parameters (source of drinking water, place of abode and type of toilet facility) did not statistically influence donor CMV status, p>0.05. CONCLUSION: Our study showed that overcrowding was the most significant factor that statistically influenced donor CMV status in Jos, Nigeria. There is, therefore, an urgent need for concerted effort to handle the problem of overcrowding in developing countries and Nigeria in particular. PMID- 19305434 TI - Medical audit of diagnostic mammographic examination at the lagos university teaching hospital (luth), Nigeria. AB - PURPOSE: We studied the performance of diagnostic mammography in predicting malignancy among patients presenting with signs and symptoms of malignancy. MATERIALS AND METHODS: 310 patients who presented to the surgical out-patients of the Lagos University Teaching Hospital with signs and symptoms of breast cancer, between January 2002 and December 2003 had mammographic evaluation with a dedicated mammography unit. The mammographic appearances were prospectively classified using the BI-RADS assessment categorisation, and histological confirmation following surgical excision was correlated with mammography. RESULTS: The overall sensitivity of mammography in the study was 82.8% (CI 78.6 - 87%) while specificity, Positive predictive value (PPV) and accuracy were 90.7% (CI 87.5-93.9%), 69.97% (CI 46.6 - 74.8%), and 89% (CI 85.5-92.5%) respectively. Multivariate analysis showed that sensitivity and specificity generally declined as breast density increased. CONCLUSION: Diagnostic mammography is useful in characterising breast lesions. This study analyses the performance of mammography and shows that the resultant specificity is high, however it may have to be used in conjunction with needle biopsy to achieve improve sensitivity and avoid unnecessary benign surgical biopsies. PMID- 19305435 TI - Pattern of genital tract abnormalities on hysterosalpingography in infertile patients in ikeja, Nigeria. AB - Structural abnormalities of the female genital tract, whether congenital or acquired could result in infertility or reproductive failure. This study retrospectively analysed the structural abnormalities found in patients who had hysterosalpingography in the Lagos State University Teaching Hospital (LASUTH), Ikeja. A descriptive analysis of structural abnormality of the uterine cavity, fallopian tube and the cervical canal in two hundred and fifty patients, who were referred from the gynaecology clinic to the Radiology Department of LASUTH, from January to December 2005, is presented. The commonest abnormality seen was tubal blockage (56.8%) whether unilateral or bilateral. Filling defects (16%) were seen within or outside the uterus, causing irregularity of its wall. These could be due to fibroids, endometrial polyps or fibrous tissue causing adhesions. Cervical canal (19.8%) abnormalities were also noted. Only two cases of congenital anomalies were seen. We found that hysterosalpingography is a relatively cheap and easy mode of diagnosing structural anomalies of the genital tract such as fibroids and tubal pathologies. PMID- 19305436 TI - Anatomical subsite and diagnostic implications of colorectal cancer in zaria (Guinea savannah)-1981-2005. AB - OBJECTIVE: To study the clinicopathological characteristics of Colorectal Cancer (CRC) in the Guinea Savannah region; identify sub site; ascertain any change in the anatomical sub-site between 1981-2005; relate tumour stage/differentiation, to age young =40 years and = 41years old patients Highlight option for diagnosis in the sub region. SETTING: There were 262 patients consisting of 125 retrospective and 137 prospective cases seen in the premier tertiary health institution in Northern Nigeria. RESULTS: The Male:Female ratio = 1.37:1, mean age 39.8 years, cluster age group 3rd-6th decade( 82.1%), patients = 40years constituted 48.2% . Main clinical features present for more than 6 months were bloody mucus discharge (83.5%), palpable rectal mass (67.9%), pain (67%), tenesmus (60.1%). Ignorance of the disease was rife amongst non-specialists, spiritualists and the patients. The left colon was eleven times more commonly affected than the right colon. The rectum was the commonest sub site (74.3%). In 2 out of every 3 patients, the tumour was digitally palpable per rectum. There was no shift in the distribution sub site in (1981-1990), and (1991-2005). 97.2% the tumours were adenocarcinoma. 68.9% patients had advanced disease Duke C or D. 3 out of every 4 patients (= 40 years age group) had advanced disease. Histological prognostic grade of tumour was inversely related to =40 years age group. Routine digital rectal examination as an investigative armamentarium,and public awareness may reduce the prevalence of advanced CRC in the low resource subregion,we advocate their use. PMID- 19305437 TI - The Prevalence and Predictors of Cigarette Smoking among Secondary School Students in Nigeria. AB - OBJECTIVES: This study was carried out to determine the prevalence of smoking among secondary school students and identify factors that influence smoking amongst them. METHODS: This descriptive and explorative study was conducted among 1,183 secondary school students, selected by multistage sampling from each of the 6 geopolitical zones in Nigeria. Data was collected by using interviewer administered structured questionnaires. RESULTS: This study recorded a lifetime smoking prevalence of 26.4% and current smoking prevalence of 17.1% among secondary school students in Nigeria. Most (82%) of the students had seen warnings against smoking and most of them were aware that it is possible for cigarette smoking to damage body organs. Unfortunately, however, seeing such warnings had no significant effect on their decision to smoke or not. The students who smoke were introduced to smoking mainly by their friends (67.4 %), and the television (13.4%). Smoking habits of the respondents were influenced by parents' educational status (p<0.05), having friends who smoke (p<0.05) and living with a smoker (p<0.05). CONCLUSION: Over a quarter of secondary school students had ever smoked. Peer pressure was the main reason cited by respondents for initiating smoking. It is suggested that our smoking prevention programmes be reviewed and appropriate health education and smoking cessation programmes be developed and implemented in order to prevent and control smoking among Nigerian students. PMID- 19305438 TI - Inadequacy of Dipstick Proteinuria in Hypertensive Pregnancy: Evidence for a change to alternatives. AB - BACKGROUND: The presence of proteinuria in pregnancy induced hypertension has been shown to increase maternal and perinatal morbidity and mortality. Hence early and accurate screening for proteinuria with prompt management holds one of the keys to reducing the morbidity and mortality associated with the disease. OBJECTIVE: To examine the options for detection of proteinuria in hypertensive pregnancy available to the obstetrician in the light of current evidence in literature. METHOD OF LITERATURE SEARCH: A review of journal articles identified through a search of the following electronic databases: Medline, Cinahl, Biomed Central and Ajol. CONCLUSION: There is overwhelming evidence that urinary dipstick results correlate poorly with 24-hour urinary protein excretion values in pregnant women with hypertension. In the light of current evidence, the random urine protein:creatinine ratio quantitatively or test strips offers the best option for screening for proteinuria in hypertension in pregnancy. PMID- 19305439 TI - Management of long bone fractures and associated injuries from gunshots at a nigerian tertiary hospital. AB - BACKGROUND: Globally, management of gunshot injuries (GSI) encompasses resuscitation, evaluation and treatment. Wound debridement is mandatory, while definitive treatment of any associated fracture varies from one part of the world to another. In developed countries aggressive modalities of early operative fixation of such fractures have produced good results. In the Nigerian environment, the approach to such fractures is controversial and cautious, with secondary operative and even non-operative methods still being predominant. OBJECTIVE: To compare operative versus non-operative methods of treatment of long bone fractures and associated injuries from gunshots in our centre. METHODS: This is a prospective study of long bone fractures and associated injuries from civilian gunshots presenting at Lagos University Teaching Hospital (LUTH), Lagos, Nigeria between 1st January 2000 and 31st December 2003. LUTH, one of the tertiary hospitals in Nigeria receives most of the multi-system trauma referrals in Lagos State, Nigeria. RESULTS: Three hundred and sixty patients with GSI were studied, 322 (89.4%) were males while 38 (10.6%) were females, giving a male: female ratio of 8.5:1. One hundred and thirty-six patients (37.8%) had long bone fractures, 94 (69.1%) on the lower limbs. All wounds were debrided and 198 (55.0%) had serial debridement. Eighty-two (60.3%) of 136 patients with fractures from G.S.I had operative treatment done secondarily and 87.5% of them achieved union. Also 42 (77.8%) of 54 patients treated non-operatively achieved union. Two hundred and thirty-four (65.0%) of the gunshot wounds healed by secondary intention, 81 (22.5%) had split thickness skin grafting and 18 (5.0%) had flap cover. CONCLUSION: This study showed that secondary operative treatment of long bone fractures in GSI produces good outcome. PMID- 19305440 TI - Changing pattern of maxillofacial fractures in a sub-urban nigerian teaching hospital. AB - OBJECTIVES: To ascertain the current pattern of facial fractures presenting in the Obafemi Awolowo University Teaching Hospital (OAUTH) and compare this with two earlier reports from the same institution. PATIENTS AND METHODS: All patients presenting with facial bone fractures at the OAUTH between January 1996 and December 2004 were included in the study. Information obtained were patients' demographics, date, cause, site and type of injury. The interval before presentation in the hospital was documented. Data was analysed and compared with two earlier studies carried out in the same institution. RESULTS: 451 patients (351 males and 100 females) presented with 632 facial bone fractures between January 1996 and December 2004. This shows a greater female involvement compared with earlier studies. Similar to earlier studies, majority of the patients (39.7%) were aged 21-30 years however, more cases occurred in the first decade of life in the present study than in the previous. Road traffic crashes remained the leading cause of fractures (74.7%), followed by fight/assaults (8.0%). There was a significantly higher prevalence of motorcycle related crashes and gunshot injuries. While the incidence of mandibular fractures declined, a statistically significant increase was observed for fractures of the middle face specifically Le Fort 1 and zygomatic complex fractures. CONCLUSIONS: Facial bone fractures appear to be on the increase in the studied location. More females and younger children are involved now. Road traffic crashes accounted for most facial bone fractures but violence related causes are on the increase. PMID- 19305441 TI - Coitally Related Traumatic Injury of the Female Genital Tract in a Nigerian Urban Setting: A-5 year review. AB - BACKGROUND: Available evidence suggests that most female lower genital tract injuries are primarily of coital origin and may result in death where prompt diagnosis and treatment is not obtained. Yet there is paucity of recent reports on this clinical entity from our setting. METHODS: This was a retrospective study of the service delivery records of patients with coital trauma seen at the University of Benin Teaching Hospital over 5 years. RESULTS: The incidence of coital trauma was 0.7% of the total gynaecological patients (3,300) seen within the study period. Majority (52.2%) of the patients were nulliparous. A significant proportion (73.9%) of the patients were single and 82.2% had primary or no formal education. Coital laceration occurred in sexual intercourse with boyfriends in 39.1% of patients and casual sex partners in 30.4% of cases. Non consensual sex was reported in 47.8% of the patients and 39.1% had consensual sex. Sex was pre-marital in 65.2% of the cases. Inadequate or lack of foreplay was a significant predisposing circumstance to coital trauma (56.5%). Pain was a presenting symptom in 60.9% of cases, and same proportion of patients had laceration > 4cm. CONCLUSION: The incidence of coitally associated trauma was low. Nulliparity, low levels of education, non-consensual and premarital sex with little or no foreplay were strongly correlated with the risk of coital trauma. Severe forms of intra-peritoneal complications were not documented in this series. Management strategies were quite adequate. PMID- 19305442 TI - Ankle arthrodesis- a 5 year review. AB - BACKGROUND: Painful arthrosis is the commonest indication for ankle arthrodesis. This procedure is infrequently undertaken in our centre - National Orthopaedic Hospital, Igbobi, Lagos (NOHIL), which is the biggest orthopaedic hospital in Nigeria. In order to determine its relevance to orthopaedic practice in Nigeria, we carried out a retrospective study on the indications, complications and functional outcome of ankle arthrodesis at the NOHIL, Nigeria. METHOD: The study was a 15-year retrospective analysis of all cases of ankle arthrodesis undertaken at NOHIL. There were 58 patients that underwent ankle arthrodesis during the period 1989 to 2003. Thirty five of these with complete records and radiographic films were included in the study. Demographic data, indications, surgical techniques, complications and outcome of treatment were analysed using SPSS version 11. RESULTS: There were 37 arthrodeses, involving 18 male and 17 female patients. The mean age was 41.4years with a mean follow-up period of 17months. The main indication for arthrodesis was posttraumatic arthrosis followed by severe acute ankle injuries. AO and Charnley compression techniques were the commonest surgical procedures employed. Eighteen (48.6%) patients had a good outcome, 10 (27.0%) patients had a fair outcome and 5 (13.5%) had a poor outcome. The outcome of the remaining 4 cases could not be ascertained as they were lost to follow up at an average of 4.6 months post surgery. CONCLUSION: The commonest indication for ankle arthrodesis was post-traumatic ankle arthrosis in this study. The use of external fixation technique was the most prevalent method of achieving arthrodesis with a fusion rate of 85%. Post surgical infection was however high but a good outcome was achieved in majority of cases. PMID- 19305443 TI - Polycythaemia vera in Nigeria. AB - BACKGROUND: There is a paucity of reports on polycythaemia vera (PV) in Nigeria. The aim of this review is to present the pattern of clinical presentation, method of diagnosis, therapeutic options and treatment outcome in the face of limited facilities. MATERIALS AND METHODS: Case notes of patients with confirmed diagnosis of PV managed at the Obafemi Awolowo University Teaching Hospitals Complex (OAUTHC), Ile-Ife, Nigeria from 1997 to 2006, were reviewed for clinical and laboratory parameters. The relative proportion of PV to other cases of haematologic cancers seen within the same period was determined. RESULTS: Seven patients, 5 males and 2 females, aged 42-70 years (median, 53 years) were studied. All the patients were symptomatic at diagnosis with the majority presenting with headaches, visual disturbances, and tinnitus. Clinical signs include conjuctival suffusion in all the patients; splenomegaly, hepatomegaly and hypertension in 3 patients (42.8%). Pruritus was uncommon (14.3%). One patient (14.3%) presented with fatal cerebrovacscular accident on admission. The average follow up period was 39.9 months, and 2 patients (28.6%) were followed up for more than 7years. Therapy consisted mainly of regular phlebotomy and low dose aspirin for suppression of thromboxane synthesis and control of thrombocytosis and erythomelalgia. PV accounts for just 0.03% of all the haematologic cancers seen. CONCLUSION: PV has a low incidence in our population and affects significantly the middle age persons. The clinical presentation consisted of headaches, visual disturbance, hypertension, and organomegaly. Treatment outcome are not different from those previously reported. The need for life-long follow up must be emphasised to patients at diagnosis. PMID- 19305444 TI - Respiratory services in New Zealand: a breath of fresh air is needed. PMID- 19305445 TI - A survey of respiratory and sleep services in New Zealand undertaken by the Thoracic Society of Australia and New Zealand (TSANZ). AB - AIMS: In 2004, the NZ Branch of the TSANZ published "Standards for Adult Respiratory and Sleep Services" on the Ministry of Health's (MoH) website.1 The aim of this survey was to evaluate each of the 21 District Health Boards' (DHBs) performance against the published standards, concentrating particularly on staffing, infrastructure, clinical support services, implementation of guidelines, quality assurance activity, and basic services (sleep, lung function, and oxygen). METHODS: Postal questionnaire survey of all DHBs in late 2006. RESULTS: All 21 DHBs responded. Only 10 of 21 DHBs were complying with the minimum standards of care. Main deficiencies in care related to: inadequate medical staffing rates, lack of quality assurance measures and insufficient laboratory testing (sleep and lung function). The lack of monitoring of such basic activities as outpatient clinic attendances, oxygen and sleep services, and the non implementation of treatment guidelines were of particular concern. Seven fold variations in prescription of assisted ventilation equipment and oxygen therapy exist across the country. CONCLUSIONS: When evaluated against minimum standards of care published in 2004, major gaps in service provision exist in New Zealand. Access to services is variable. There is a lack of national leadership and insufficient regional organisation leading to large gaps in service provision of even basic respiratory services. Immediate changes to the current service provision structures are required. PMID- 19305446 TI - Live donor liver transplantation in New Zealand: a report on the first 20 cases. AB - BACKGROUND: Liver transplantation (LT) is established treatment for adults and children with acute or chronic liver failure, however there are insufficient donor organs to meet demand and 14% of New Zealand patients have died waiting or were de-listed due to deterioration whilst on the waiting list. Live donor liver transplantation (LDLT) offers an alternative graft source that enables timely transplantation, but also carries the risk of morbidity and mortality for the donor. AIM: To report the initial experience with LDLT in New Zealand. METHODS: Review of donor and recipient outcomes for the first 20 cases. RESULTS: 129 potential live liver donors were assessed for 68 recipients. Donors were evaluated according to a multi-step protocol including independent donor advocacy. Twenty LDLT were performed on 7 adults and 13 paediatric recipients using 5 right lobe, 2 extended left lobe, 2 left lobe, and 11 left lateral section grafts. Five donors (25%) experienced postoperative complications, none of which were life-threatening. Four recipients had acute liver failure and 16 had chronic liver disease including one retransplant. There was a high rate of recipient biliary complications (40%) but graft and recipient survival is 100% to date. CONCLUSION: LDLT has been successfully introduced in New Zealand with good donor and recipient outcomes. PMID- 19305447 TI - The readmission rate as an indicator of the quality of elective surgical inpatient care for the elderly in New Zealand. AB - AIM: To conduct a hypothesis-raising and descriptive study of the rate of readmission/death for patients aged over 64 years as a measure of the quality of inpatient care, for elective surgical procedures between 2001 and 2004. METHODS: Data from the New Zealand Health Information Service was used to calculate an annual rate for patients aged 65 years or over between July 2000 and July 2004 who were readmitted or deceased within 30-days of discharge following an: elective transurethral prostatectomy, laparoscopic cholecystectomy, knee arthroplasty, hip arthroplasty, or inguinal hernia repair. RESULTS: It is probable that the risk of readmission/death within 30-days of discharge ('RoD') rose 13% (95% CI of increase: 0%, 27%) from 7.5% in 2001/2002 to 8.5% in 2003/2004. The risk of RoD was greater among patients aged 80 years or over (RR 1.38, 95% CI 1.26, 1.51), males (RR 1.26, 95% CI 1.12, 1.41), and NZ Maori (RR 1.6, 95% CI 1.2, 2.3). CONCLUSION: There is evidence for a probable increase in the rate of RoD between 2001 and 2004, and its relative risk varied with gender, age, and ethnicity. However, this study was not able to control for potential confounders (length-of-stay, casemix, or comorbidities) which may affect the estimated result. Ongoing research is recommended to explore the use of RoD rate as an indicator of health services quality and consider whether this rate is increasing, despite health system quality interventions. In addition, further investigation is needed to evaluate the quality of hospital care in New Zealand with respect to ethnicity, age, and gender. PMID- 19305448 TI - Lymph node infarction and its association with lymphoma: a short series and literature review. AB - AIMS: Lymph node infarction is an extremely rare phenomenon. An infarcted lymph node can proceed, or occur simultaneously with, lymphoma. In the present study we review the literature on lymph node infarction and describe our experience of such cases. METHODS: Six cases of lymph nodes with a diagnosis of lymph node infarction were archived from the records of the Pathology Department. Clinical information of the patients was obtained from the case records. Haematoxylin andeosin stained sections were reviewed. RESULTS: A diagnosis of non-Hodgkin lymphoma was made in 5 of the 6 patients. The diagnosis was made concurrent to the lymph node infarction in 2 cases while in 3 cases the diagnosis was made on subsequent follow up biopsies. The aetiology of lymph node infarction could not be established in one case. CONCLUSION: The pathologist should be cautious when examining an infarcted lymph node. Though all patients might not develop lymphoma, they require a close follow-up and repeat biopsies to detect the subsequent development of lymphoma. PMID- 19305449 TI - Sentinel lymph node biopsy experience in Taranaki: a prospective audit in a provincial New Zealand hospital. AB - AIM: Sentinel lymph node biopsy has been rapidly incorporated into the management of early stage invasive breast cancer. The aim of this study was to review the adoption of sentinel lymph node biopsy at a provincial centre in New Zealand and compare markers of performance against established standards. METHODS: The Taranaki Breast Database was created in 2002 and prospectively records data from all breast cancer patients in the Taranaki area. Data on all patients undergoing sentinel lymph node biopsy were retrieved and the results reviewed. RESULTS: Between October 2002 and August 2007, 152 sentinel lymph node biopsies were undertaken in 151 patients. The initial 49 patients (training set) also underwent routine axillary clearance as part of an initial audit on the accuracy of sentinel lymph node biopsy. A sentinel node was identified in 97% of patients (93% including the training set) and a mean of two nodes per biopsy were removed. Metastatic nodal disease was identified in 40 of 152 (26%) of biopsies of which nine were micrometastases. In the training set there was a false negative rate for nodal spread of 5% (two of 40) and a 92% negative predictive value. CONCLUSIONS: The performance of sentinel lymph node biopsy in Taranaki is comparable to international centres. Adoption of this technique as routine may spare many Taranaki women the morbidity of axillary clearance, without jeopardising safety. PMID- 19305450 TI - Functional outcome of surgery for fractures of the ankle. AB - BACKGROUND: Ankle fractures are a common injury treated in New Zealand with variable functional outcomes. METHODS: Details of all patients undergoing operative fixation of malleolar fractures at Hutt Hospital were obtained from theatre databases for a 12-month period. These patients were then followed up after 1 year for clinical review and asked to complete a questionnaire. RESULTS: Sixty-two patients enrolled in the study: 27 men and 35 women. Average OMA score 16 months after surgery of 79.4. Weber A fractures averaged ankle function score 90.0, Weber B fractures 80.8, and Weber C fractures 76.3. Two patients (3%) reported 'poor' results, 11 (18%) reported 'fair' results, 29 (47%) gained 'good' results, and 20 (32%) attained 'excellent' results. Functional outcome was worse in the elderly. CONCLUSION: Patients who sustain ankle fractures have significant functional impairment even after 1 year. PMID- 19305451 TI - The validity of readmission rate as a marker of the quality of hospital care, and a recommendation for its definition. AB - AIM: To perform a review of relevant literature regarding the use of readmission rate as a marker of the quality of hospital care, summarise its validity, and recommend a definition for its use. METHODS: Literature search was performed on the Embase and Medline databases, with relevant articles extracted and reviewed. CONCLUSIONS: Readmission rate as a marker of the quality of hospital care has been used both internationally and nationally, although its validity has only been partially substantiated. While prone to confounding, it remains a valuable indicator due to its ease of collection and its ability to be able to be combined with other variables. Although the definition of readmission rate varies in the literature, it may be defined as 'the number of patients who experienced unintended, acute readmission or death within 30-days of discharge from the index admission, divided by the total number of patients discharged alive within the reference period'. PMID- 19305452 TI - Acute care surgery: can New Zealand afford to wait? AB - Until about two decades ago, the provision of emergency surgery was implicitly linked to all aspects of surgical care in all surgical specialties. While this remains true in the smaller surgical specialties, in the larger specialties the development of subspecialisation has eroded the comprehensive nature of acute care provision. In general surgery, the numerically largest of the nine surgical specialties, the greatest challenges in provision of acute care exist. Availability of appropriately trained general surgeons to deliver generic acute and trauma care has reached crisis point. An attempt is being made in the Western World to remedy these problems. Recognising that it is increasingly difficult to span the knowledge and skill mix necessary to manage all aspects of acute care in general surgery as well as a subspecialty practice, the concept of acute care surgery has been born. To look at the status of acute care surgery in Australia and New Zealand, we conducted a PubMed search on all articles matching the words emergency, acute care, and general surgery, and reviewed any papers relevant to Australasia. Of the 270 papers found, 4 papers were relevant to Australia and New Zealand. These studies outline the advantages of an acute care model in dealing with emergency surgery and delineate few disadvantages. Whether a new training paradigm, "acute care surgery" will benefit patients, the health services, and the surgeons in New Zealand and elsewhere remains to be seen. Allowing the current trend towards inadequate numbers and training of surgeons to deliver acute care to continue is unacceptable. PMID- 19305453 TI - Multiphasic cerebral demylination. PMID- 19305454 TI - A rare complication of wireless capsule endoscope. PMID- 19305455 TI - Medical image. Massive gastric dilatation secondary to a binge episode in bulimia nervosa. PMID- 19305456 TI - Medical image. Tietze's syndrome. PMID- 19305457 TI - Overshooting the mark: subclinical hyperthyroidism secondary to excess thyroid hormone treatment may be more prevalent than we realise. PMID- 19305458 TI - Two-dimensional cell parameters of twisted nematic liquid crystal with an amplitude-sensitive heterodyne ellipsometer. AB - To be compared with the wavelength modulation technique for measuring two dimensional (2D) cell parameters of a twisted nematic liquid crystal (TN-LC), we propose an amplitude-sensitive heterodyne ellipsometer (ASHE) of a single wavelength that is able to characterize TN-LC in 2D quantitatively. A quarter wave plate (QWP) is rotated continuously in this setup to modulate the polarization state of the incident laser beam to obtain the amplitude ratio of the S and P waves versus the rotation angle of the QWP. Thus the cell parameters, including the twisted angle Phi, untwisted phase retardation Gamma, rubbing direction angle alpha, and cell gap d, of a TN-LC cell are obtained simultaneously by best fitting the detected amplitude ratio with a prediction based on the transfer matrix of TN-LC cell. 2D distributions of (Phi,Gamma,alpha,d) are then obtained either by scanning the TN-LC cell or by using a CCD camera for high-speed measurement. In this experiment, the stability of the amplitude-ratio measurement of the proposed ASHE was 0.5%. The goal is to integrate the rotating elliptical wave plate with the TN-LC cell in a heterodyne ellipsometer to obtain cell parameters at amplitude sensitivity. This increases not only the sensitivity of the measurement but also the possibility of extending the 2D distribution of cell parameters in real time. PMID- 19305459 TI - Reduction of polarization related effects in superimposed fiber Bragg gratings. AB - Fiber Bragg gratings, and more specifically, superimposed fiber Bragg gratings (SIFBGs), are attractive commercial solutions for several multiband telecommunication applications. However, as a part of a telecommunication system, the polarization dependent properties present in SIFBGs due to the fabrication process dramatically limit their possible implementation in high bit rate optical communications. The development of techniques for the reduction of differential group delay (DGD) and the polarization dependent loss (PDL) in system components is then crucial. We present a simple method to reduce the DGD and the PDL induced during the fabrication of SIFBGs. The proposed fabrication method consists of irradiating the fiber core from different well controlled directions depending on the total number of expositions. We theoretically predict and experimentally demonstrate a reduction of the DGD and the PDL after each illumination. PMID- 19305460 TI - Optical force model based on sequential ray tracing. AB - We discuss how information available from ray-tracing techniques can be used to calculate optical forces and torques on particles. A general ray-trace computer code is augmented with the polarization and irradiance distributions of the illumination and Fresnel surface coefficients to give a reasonably accurate prediction of interaction with large particles out of the focal plane. Calculations of trapping location versus nonuniform illumination conditions are compared with an experiment. Other example calculations include trapping a hemispherical lens and a two-particle trap. PMID- 19305461 TI - Measurement of period difference in grating pair based on compensation analysis of phase difference between diffraction beams. AB - We propose an experimental method for measuring the period difference in a grating pair. The method uses the relationship between the period difference and the phase difference of diffraction beams of the grating pair when gratings are translated along the grating-vector direction. The effect of the roll angular deviation of gratings can be eliminated by phase compensation with +1st- and -1st order diffraction beams. Far-field intensity patterns of diffraction beams are monitored to adjust the relative attitude of gratings. For a pair of gratings with periods of approximately 0.674 microm, the period difference was measured to be 0.4434 nm with an expanded uncertainty (k=2) of 0.0113 nm. PMID- 19305462 TI - Effect of water impurity in CsLiB6O10 crystals on bulk laser-induced damage threshold and transmittance in the ultraviolet region. AB - We investigate the effect of water impurity in a CsLiB(6)O(10) (CLBO) crystal on the ultraviolet properties of the bulk laser-induced damage threshold (LIDT) and transmittance. The water impurity was eliminated by heating the CLBO sample with dimensions of 5 mm x 5 mm x 10 mm at 150 degrees C in an ambient atmosphere and subsequently in a dry atmosphere. The bulk LIDT of the sample after heat treatment improved by about 1.6-fold compared with that before heat treatment. PMID- 19305463 TI - Snapshot polarimeter fundus camera. AB - A snapshot imaging polarimeter utilizing Savart plates is integrated into a fundus camera for retinal imaging. Acquired retinal images can be processed to reconstruct Stokes vector images, giving insight into the polarization properties of the retina. Results for images from a normal healthy retina and retinas with pathology are examined and compared. PMID- 19305464 TI - Deep UV light generation by a fiber/bulk hybrid amplifier at 199 nm. AB - A high-pulse-repetition-frequency (PRF) pulsed light source in the deep ultraviolet region has been realized by a multiple wavelength conversion technique using a hybrid fiber/bulk amplifier system. Output of 199 nm with a power of 50 mW was achieved at 2.4 MHz PRF. The 1 microm amplifier consisted of a Yb-doped fiber amplifier and a Nd-doped YVO(4) amplifier. A 1.5 microm fiber master-oscillator power amplifier was employed as the other fundamental source. The amplifiers exhibited good amplification properties in pulse energy, polarization extinction ratio, and spectrum for nonlinear wavelength conversion. PMID- 19305465 TI - Experiments of a grating light modulator for projection display applications. AB - A micromechanical optical modulator called a grating light modulator (GLM) with electrostatic actuation, fabricated by the conventional process, is described. The GLM is operated by the interaction of a stationary reflector and a movable sliding grating. The GLM is a micromechanical phase grating. The phase difference is determined by the slide of the movable part (upper grating), allowing different diffraction patterns of reflected light. In addition, 100% modulation in the first/zero order can act as an optical switch and produce bright or dark pixels in a projection display system. Built using microelectromechanical system technology, and designed to be manufactured using mainstream integrated circuit fabrication technology, a 256-pixel GLM array is fabricated. We also describe projection display applications of a GLM as a high efficiency spatial light modulator. The optimized display system with a GLM is theoretically discussed in detail. The single-color and three-color experimental results indicate that the GLM technology has characteristics that make it suitable for projection display applications. PMID- 19305466 TI - Improvement and analysis of a micro Raman probe. AB - A micro Raman probe (MRP) with a 600 microm diameter, which we previously reported as the narrowest achieved to date, was further improved by introducing high-quality optical filters and a collecting lens at the tip. We fabricated the MRP with a high collection efficiency, a wider collection wavelength, and a high signal-to-noise ratio. We compared two types of probes: one with a lens-tipped end and one with a flat tip. We experimentally tested the performance of these MRPs to evaluate the detection properties defined by parameters such as the optical purity against inherent Raman background noise due to optical fibers, the sensitivity, and the viewing area. Finally, we demonstrated their effectiveness in measurements of standard Raman samples and applied them to measurements of plastic and human skin samples in situ. PMID- 19305467 TI - Optical phase and intensity modulation from a rotating optical flat: effect on noise in degree of polarization measurements. AB - A spectral analysis of the electrical signal from the detector in degree of polarization (DOP) measurements that use a rotating polarizer shows base band frequencies that create a noise floor. The noise floor arises from phase and intensity modulation of the optical field owing to the varying thickness and transmission of the rotating optical polarizer in the DOP apparatus. A physical model is presented for the noise floor arising from the phase and intensity modulation, and a calibration procedure including configuration guidelines is provided to minimize the effects of the unwanted modulations. PMID- 19305468 TI - Active quenching circuit for single-photon detection with Geiger mode avalanche photodiodes. AB - In this paper a novel construction of an active quenching circuit intended for single-photon detection is presented, along with a few original methods for its evaluation. The circuit has been combined with a standard avalanche photodiode C30902S to form a single-photon detector. This detector has a dead time of 39 ns, maximum random counting frequency of 14 MHz, small afterpulsing probability, an estimated peak detection efficiency of over 20%, and a dark count rate of less than 100 Hz. This simple and robust active quenching circuit can be built from off-the-shelf electronic components and is presented with the detailed schematic diagram. PMID- 19305469 TI - Annealing study of carrier concentration in gradient-doped GaAs/GaAlAs epilayers grown by molecular beam epitaxy. AB - We measured the carrier concentration distribution of gradient-doped GaAs/GaAlAs epilayers grown by molecular beam epitaxy before and after annealing at 600 degrees C, using electrochemical capacitance voltage profiling, to investigate the internal variation of transmission-mode GaAs photocathodes arising from the annealing process. The results show that the carrier concentration increased after annealing. As a result, the total band-bending energy in the gradient-doped GaAs emission layer increased by 25.24% after annealing, which improves the photoexcited electron movement toward the surface. On the other hand, the annealing process resulted in a worse carrier concentration discrepancy between the GaAs and the GaAlAs, which causes a lower back interface potential barrier, decreasing the amount of high-energy photoelectrons. PMID- 19305470 TI - Fast beam steering with a ferroelectric-liquid-crystal optical phased array. AB - We demonstrate fast, efficient beam steering using a single 1x32 analog ferroelectric liquid crystal (FLC) spatial light modulator. A high-tilt FLC material with 82 degrees optic-axis switching provides, in a reflective-mode device with a passive quarter-wave retarder between a half-wave FLC layer and a mirror, 91% of full 0-2pi phase modulation. Electronic drive based on applied charge gives 200 micros response-time analog modulation. PMID- 19305471 TI - Atomic layer deposition of Al2O3 and TiO2 multilayers for applications as bandpass filters and antireflection coatings. AB - Al(2)O(3) and TiO(2) thin films have been deposited on Si wafers, quartz, BK7 glass, and polycarbonate substrates by atomic layer deposition (ALD). The refractive indices and growth rates of the materials have been determined by spectroscopic ellipsometry and transmission electron microscopy. The influence of substrate temperature and precursor on the refractive indices has been investigated. The refractive index of TiO(2) significantly increases with temperature, whereas the Al(2)O(3) films are temperature insensitive. The films deposited using H(2)O(2) as oxygen source show a slightly higher refractive index than the films prepared with H(2)O. Multilayer narrow-bandpass filters and broadband antireflective coatings have been designed and produced by ALD. PMID- 19305472 TI - Balanced interferometric system for stability measurements. AB - We describe two different, double-sided interferometer designs for measuring material stability. Both designs are balanced interferometers where the only optical path difference is the sample and the reference beams are located within the interferometer. One interferometer is a double-pass design, whereas the other is a single-pass system. Based on a tolerancing analysis, the single-pass system is less susceptible to initial component misalignment and motions during experiments. This single-pass interferometer was tested with an 86 nm thin-film silver sample for both short-term repeatability and long-term stability. In 66 repeatability tests of 30 min each, the mean measured drift rate was less than 1 pm/h rms. In two long-term tests (>9 h), the mean drift rate was less than 1.1 pm/h, which shows good agreement between the short- and long-term measurements. In these experiments, the mean measured length change was 2 nm rms. PMID- 19305473 TI - Optical design and multiobjective optimization of miniature zoom optics with liquid lens element. AB - We propose an optical design for miniature 2.5x zoom fold optics with liquid elements. First, we reduce the volumetric size of the system. Second, this newly developed design significantly reduces the number of moving groups for this 2.5x miniature zoom optics (with only two moving groups compared with the four or five groups of the traditional zoom lens system), thanks to the assistance of liquid lens elements in particular. With regard to the extended optimization of this zoom optics, relative illuminance (RI) and the modulation transfer function (MTF) are considered because the more rays passing through the edge of the image, the lower will be the MTF, at high spatial frequencies in particular. Extended optimization employs the integration of the Taguchi method and the robust multiple criterion optimization (RMCO) approach. In this approach, a Pareto optimal robust design solution is set with the aid of a certain design of the experimental set, which uses analysis of variance results to quantify the relative dominance and significance of the design factors. It is concluded that the Taguchi method and RMCO approach is successful in optimizing the RI and MTF values of the fold 2.5x zoom lens system and yields better and more balanced performance, which is very difficult for the traditional least damping square method to achieve. PMID- 19305474 TI - Measuring the volume of a fluid in a diamond anvil cell using a confocal microscope. AB - Confocal microscopy is a potentially powerful technique for obtaining equation-of state (EOS) data for fluids in a diamond anvil cell. Unlike conventional microscopy, a confocal microscope scans the cell in three dimensions. From the intensity profile of the reflected laser light, we calculated the index of refraction and optical thickness of the sample contained in the cell. These measurements, combined with the cross-sectional area of the sample, enabled us to calculate the volume. As a test of the experimental technique and analysis, we produced a pressure-volume curve for liquid water at 300 K. The results agree with published EOS data within experimental error. PMID- 19305476 TI - Quantum random number generator using photon-number path entanglement. AB - We report a quantum random number generator based on the photon-number-path entangled state that is prepared by means of two-photon quantum interference at a beam splitter. The randomness in our scheme is truly quantum mechanical in origin since it results from the projection measurement of the entangled two-photon state. The generated bit sequences satisfy the standard randomness test. PMID- 19305477 TI - Noise effect in an improved conjugate gradient algorithm to invert particle size distribution and the algorithm amendment. AB - In general, model-independent algorithms are sensitive to noise during laser particle size measurement. An improved conjugate gradient algorithm (ICGA) that can be used to invert particle size distribution (PSD) from diffraction data is presented. By use of the ICGA to invert simulated data with multiplicative or additive noise, we determined that additive noise is the main factor that induces distorted results. Thus the ICGA is amended by introduction of an iteration step adjusting parameter and is used experimentally on simulated data and some samples. The experimental results show that the sensitivity of the ICGA to noise is reduced and the inverted results are in accord with the real PSD. PMID- 19305478 TI - Simultaneous measurement of linear and transverse displacements by laser self mixing. AB - We present a contactless optical sensor based on the laser-self-mixing effect for real-time measurement of linear and transverse displacements of a moving stage. The sensor is able to measure linear displacements of up to 400 mm along the main optical axis while simultaneously estimating straightness and flatness deviations up to 1 mm. The sensor exploits two identical coplanar nonparallel self-mixing interferometers and requires only one reference plane. The reduction in the number of optical elements allowed by the self-mixing configuration and the intrinsic stiffness of the adopted geometry result in a compact, low-cost, and easy-to-align setup. PMID- 19305479 TI - Evidence-based or evidence-biased: the need to re-appraise and re-align levels of information with stakeholder values. PMID- 19305480 TI - Neurogliamodulation. PMID- 19305481 TI - Obama health care for all Americans: practical implications. AB - Rapidly rising health care costs over the decades have prompted the application of business practices to medicine with goals of improving the efficiency, restraining expenses, and increasing quality. Average health insurance premiums and individual contributions for family coverage have increased approximately 120% from 1999 to 2008. Health care spending in the United States is stated to exceed 4 times the national defense, despite the wars in Iraq and Afghanistan. The U.S. health care system has been blamed for inefficiencies, excessive administrative expenses, inflated prices, inappropriate waste, and fraud and abuse. While many people lack health insurance, others who do have health insurance allegedly receive care ranging from superb to inexcusable. In criticism of health care in the United States and the focus on savings, methodologists, policy makers, and the public in general seem to ignore the major disadvantages of other global health care systems and the previous experiences of the United States to reform health care. Health care reform is back with the Obama administration with great expectations. It is also believed that for the first time since 1993, momentum is building for policies that would move the United States towards universal health insurance. President Obama has made health care a central part of his domestic agenda, with spending and investments in Children's Health Insurance Program (CHIP), American Recovery and Reinvestment Act of 2009, and proposed 2010 budget. It is the consensus now that since we have a fiscal emergency, Washington is willing to deal with the health care crisis. Many of the groups long opposed to reform, appear to be coming together to accept a major health care reform. Reducing costs is always at the center of any health care debate in the United States. These have been focused on waste, fraud, and abuse; administrative costs; improving the quality with health technology information dissemination; and excessive regulations on the health care industry in the United States. Down payment on health care reform, American Recovery and Reinvestment Act, and CHIP include many provisions to reach towards universal health care. PMID- 19305482 TI - Systematic review of cervical discography as a diagnostic test for chronic spinal pain. AB - BACKGROUND: Chronic neck pain represents a significant public health problem. Despite high prevalence rates, there is a lack of consensus regarding the causes or treatments for this condition. Based on controlled evaluations, the cervical intervertebral discs, facet joints, and atlantoaxial joints have all been implicated as pain generators. Cervical provocation discography, which includes disc stimulation and morphological evaluation, is often used to distinguish a painful disc from other potential sources of pain. Yet in the absence of validation and controlled outcome studies, the procedure remains mired in controversy. STUDY DESIGN: A systematic review of the cervical discography literature. OBJECTIVE: To evaluate the validity and usefulness of cervical provocation discography in managing and diagnosing discogenic pain by means of a systematic review. METHODS: Following a comprehensive search of the literature, selected studies were subjected to a modified Agency for Healthcare Research and Quality (AHRQ) diagnostic accuracy evaluation. Qualitative analysis was conducted using 5 levels of evidence, ranging from Level I to III with 3 subcategories in Level II. The rating scheme was modified to evaluate the diagnostic accuracy. RESULTS: A systematic review of the literature demonstrated that cervical discography plays a significant role in selecting surgical candidates and improving outcomes, despite concerns regarding the false-positive rate, lack of standardization, and assorted potential confounding factors. Based on the studies utilizing the International Association for the Study of Pain (IASP) criteria, the data show a prevalence rate ranging between 16% and 20%. Based on the 3 studies that utilized IASP criteria during the performance of cervical discography, the evidence derived from studies evaluating the diagnostic validity of the procedure, the indicated level of evidence is Level II-2 based on modified U.S. Preventive Services Task Force (USPSTF) criteria. LIMITATIONS: Limitations include a paucity of literature, poor methodologic quality, and very few studies performed utilizing IASP criteria. CONCLUSION: Cervical discography performed according to the IASP criteria may be a useful tool for evaluating chronic cervical pain, without disc herniation or radiculitis. Based on a modified AHRQ accuracy evaluation and USPSTF level of evidence criteria, this systematic review indicates the strength of evidence as Level II-2 for diagnostic accuracy of cervical discography. PMID- 19305483 TI - Systematic review of diagnostic utility and therapeutic effectiveness of cervical facet joint interventions. AB - BACKGROUND: Chronic, recurrent neck pain is common and is associated with high pain intensity and disability, which is seen in 14% of the adult general population. Controlled studies have supported the existence of cervical facet or zygapophysial joint pain in 36% to 67% of these patients. However, these studies also have shown false-positive results in 27% to 63% of the patients with a single diagnostic block. There is also a paucity of literature investigating therapeutic interventions of cervical facet joint pain. STUDY DESIGN: A systematic review of cervical facet joint interventions. OBJECTIVE: To evaluate the accuracy of diagnostic facet joint nerve blocks and the effectiveness of cervical facet joint interventions. METHODS: Medical databases and journals were searched to locate all relevant literature from 1966 through December 2008 in the English language. A review of the literature of the utility of facet joint interventions in diagnosing and managing facet joint pain was performed according to the Agency for Healthcare Research and Quality (AHRQ) criteria for diagnostic studies and observational studies and the Cochrane Musculoskeletal Review Group criteria as utilized for interventional techniques for randomized trials. LEVEL OF EVIDENCE: The level of evidence was defined as Level I, II, or III based on the quality of evidence developed by the U.S. Preventive Services Task Force (USPSTF). OUTCOME MEASURES: For diagnostic interventions, studies must have been performed utilizing controlled local anesthetic blocks which achieve at minimum 80% relief of pain and the ability to perform previously painful movements. For therapeutic interventions, the primary outcome measure was pain relief (short term relief up to 6 months and long-term relief greater than 6 months) with secondary outcome measures of improvement in functional status, psychological status, return to work, and reduction in opioid intake. RESULTS: Based on the utilization of controlled comparative local anesthetic blocks, the evidence for the diagnosis of cervical facet joint pain is Level I or II-1. The indicated evidence for therapeutic cervical medial branch blocks is Level II-1. The indicated evidence for radiofrequency neurotomy in the cervical spine is Level II 1 or II-2, whereas the evidence is lacking for intraarticular injections. LIMITATIONS: A systematic review of cervical facet joint interventions is hindered by the paucity of published literature and lack of literature for intraarticular cervical facet joint injections. CONCLUSIONS: The evidence for diagnosis of cervical facet joint pain with controlled comparative local anesthetic blocks is Level I or II-1. The indicated evidence for therapeutic facet joint interventions is Level II-1 for medial branch blocks, and Level II-1 or II-2 for radiofrequency neurotomy. PMID- 19305484 TI - Systematic review of intrathecal infusion systems for long-term management of chronic non-cancer pain. AB - BACKGROUND: Disability, societal, and health impact of chronic intractable pain secondary to various failed therapies is a major issue. As advanced therapy, implantable therapies, which include intrathecal devices and spinal cord stimulation systems, are frequently used in managing chronic intractable pain. Thus, continuous infusion of intrathecal medication is one of the methods used for the control of chronic, refractory, cancer, and non-cancer pain. However, despite the high costs of chronic non-cancer pain, it has been claimed that there is a lack of evidence for intrathecal infusion systems and the cost effectiveness of these systems has been questioned in improving pain and function. STUDY DESIGN: A systematic review of intrathecal infusion devices for chronic non cancer pain. OBJECTIVE: To determine the efficacy, utilization, safety, and complications associated with the use of intrathecal infusion devices for long term management of chronic non-cancer pain. METHODS: Literature search was performed through EMBASE, Medline, Cochrane databases, and systematic reviews identified from 1966 to December 2008. Studies were then reviewed and assessed using the Agency for Healthcare Research and Quality (AHRQ) criteria for observational studies and the Cochrane Musculoskeletal Review Group criteria for randomized trials. The level of evidence was determined using 5 levels of evidence, ranging from Level I to III with 3 subcategories in Level II, based on the quality of evidence developed by the U.S. Preventive Services Task Force (USPSTF). OUTCOME MEASURES: The primary outcome measure was pain relief (short term relief < or = one-year and long-term > one-year). Secondary outcome measures of improvement in functional status, psychological status, return to work, and reduction in opioid intake were also utilized. RESULTS: The level of evidence for intrathecal infusion systems indicated either Level II-3 or Level III (limited) based on U.S. Preventive Services Task Force (USPSTF) criteria. LIMITATIONS: The limitations of this study include the paucity of literature, lack of quality evidence, and lack of randomized trials. CONCLUSION: This systematic review illustrates Level II-3 or Level III (limited) evidence for intrathecal infusion systems for long-term relief in chronic non-cancer pain. PMID- 19305485 TI - Systematic review of percutaneous adhesiolysis and management of chronic low back pain in post lumbar surgery syndrome. AB - BACKGROUND: Post lumbar surgery syndrome or failed back surgery syndrome with persistent pain continues to increase over the years. The speculated causes of post lumbar laminectomy syndrome include acquired stenosis, epidural fibrosis, arachnoiditis, radiculopathy, and recurrent disc herniation. Epidural fibrosis may account for as much as 20% to 36% of all cases of failed back surgery syndrome. Percutaneous epidural adhesiolysis has been employed in interventional pain management in the treatment of chronic, refractory low back and lower extremity pain after back surgery. STUDY DESIGN: A systematic review of randomized trials and observational studies. OBJECTIVE: To evaluate the effectiveness of percutaneous adhesiolysis in managing chronic low back and lower extremity pain due to post lumbar surgery syndrome. METHODS: A comprehensive literature search was conducted utilizing electronic databases, as well as systematic reviews and cross references from 1966 through December 2008. The quality of individual articles used in this analysis was assessed by modified Cochrane review criteria for randomized trials and the Agency for Healthcare Research and Quality (AHRQ) criteria for assessment of observational studies. Clinical relevance was evaluated using 5 questions according to the criteria recommended by the Cochrane Review Back Group. Analysis was conducted using 5 levels of evidence, ranging from Level I to III, with 3 subcategories in Level II. OUTCOME PARAMETERS: The primary outcome measure was pain relief (short-term relief of at least 6 months and long-term relief of more than 6 months). Secondary outcome measures were improvement in functional status, psychological status, return to work, and change in opioid intake. RESULTS: Of the 13 studies considered for inclusion, 3 randomized trials and 4 observational studies met the inclusion criteria for methodologic quality assessment and evidence synthesis based on methodologic quality scores of 50 or more. Evidence of percutaneous adhesiolysis in the management of chronic low back pain in post-lumbar surgery syndrome is Level I to Level II-1, with evidence derived from 3 randomized trials. LIMITATIONS: There is a paucity of efficacy and pragmatic trials. No trials have been published after 2006. CONCLUSION: The indicated level of evidence for percutaneous adhesiolysis is Level I or II-1 based on the US Preventative Services Task Force (USPSTF) criteria. PMID- 19305486 TI - Spinal cord stimulation for patients with failed back surgery syndrome: a systematic review. AB - BACKGROUND: Failed back surgery syndrome is common in the United States. Management of post lumbar surgery syndrome with multiple modalities includes interventional techniques, resulting in moderate improvement, leaving a proportion of patients in intractable pain. The systematic reviews of long-term benefits and risks of spinal cord stimulation (SCS) for patients with failed back surgery syndrome showed limited to moderate evidence and cost effectiveness. However, with the exponential increase in surgery in the United States, spinal cord implants are also increasing. Thus, the discussion continues with claims of lack of evidence on one hand and escalating increases in utilization on the other hand. STUDY DESIGN: A systematic review of SCS in patients with failed back surgery syndrome. OBJECTIVES: This systematic review is undertaken to examine the evidence from randomized controlled trials (RCTs) and observational studies to evaluate the effectiveness of SCS in post lumbar surgery syndrome and to demonstrate clinical and cost effectiveness. METHODS: Review of the literature was performed according to the Cochrane Musculoskeletal Review Group Criteria as utilized for interventional techniques for randomized trials and the Agency for Healthcare Research and Quality (AHRQ) criteria for observational studies. The 5 levels of evidence were classified as Level I, II, or III with 3 subcategories in Level II based on the quality of evidence developed by the U.S. Preventive Services Task Force (USPSTF). Data sources included relevant literature of the English language identified through searches of PubMed and EMBASE from 1966 to December 2008, and manual searches of bibliographies of known primary and review articles. OUTCOME MEASURES: The primary outcome measure was pain relief (short term relief < or = one-year and long-term > one-year). Secondary outcome measures of improvement in functional status, psychological status, return to work, and reduction in opioid intake were utilized. RESULTS: The indicated evidence is Level II-1 or II-2 for long-term relief in managing patients with failed back surgery syndrome. LIMITATIONS: The limitations of this review included the paucity and heterogeneity of the literature. CONCLUSION: This systematic review evaluating the effectiveness of SCS in relieving chronic intractable pain of failed back surgery syndrome indicated the evidence to be Level II-1 or II-2 for clinical use on a long-term basis. PMID- 19305487 TI - Evaluation of sacroiliac joint interventions: a systematic appraisal of the literature. AB - BACKGROUND: The sacroiliac joint has been implicated as a source of low back and lower extremity pain. There are no definite historical, physical, or radiological features that can definitively establish a diagnosis of sacroiliac joint pain. Based on the present knowledge, an accurate diagnosis is made only by controlled sacroiliac joint diagnostic blocks. The sacroiliac joint has been shown to be a source of pain in 10% to 27% of suspected patients with chronic low back pain utilizing controlled comparative local anesthetic blocks. STUDY DESIGN: A systematic review of diagnostic and therapeutic sacroiliac joint interventions. OBJECTIVE: To evaluate the accuracy of diagnostic sacroiliac joint interventions and the utility of therapeutic sacroiliac joint interventions. METHODS: The literature search was carried out by searching the databases of PubMed, EMBASE, and Cochrane reviews. Methodologic quality assessment of included studies was performed using the Agency for Healthcare Research and Quality (AHRQ) methodologic quality criteria for diagnostic accuracy and observational studies, whereas randomized trials were evaluated utilizing the Cochrane review criteria. Only studies with scores of 50 or higher were included for assessment. Level of evidence was based on the U.S. Preventive Services Task Force (USPSTF) criteria. OUTCOME MEASURES: For diagnostic interventions, the outcome criteria included at least 50% pain relief coupled with a patient's ability to perform previously painful maneuvers with sustained relief using placebo-controlled or comparative local anesthetic blocks. For therapeutic purposes, outcomes included significant pain relief and improvement in function and other parameters. Short-term relief for therapeutic interventions was defined as 6 months or less, whereas long-term effectiveness was defined as greater than 6 months. RESULTS: The indicated level of evidence is II-2 for the diagnosis of sacroiliac joint pain utilizing comparative, controlled local anesthetic blocks. The prevalence of sacroiliac joint pain is estimated to range between 10% and 38% using a double block paradigm in the study population. The false-positive rate of single, uncontrolled, sacroiliac joint injections is 20% to 54%. The evidence for provocative testing to diagnose sacroiliac joint pain is Level II-3 or limited. For radiofrequency neurotomy the indicated evidence is limited (Level II-3) for short- and long-term relief. LIMITATIONS: The limitations of this systematic review include the paucity of literature evaluating the role of both diagnostic and therapeutic interventions and widespread methodological flaws. CONCLUSIONS: The indicated evidence for the validity of diagnostic sacroiliac joint injections is Level II-2. The evidence for the accuracy of provocative maneuvers in the diagnosing of sacroiliac joint pain is limited (Level II-3). The evidence for radiofrequency neurotomy is also limited (Level II-3). PMID- 19305488 TI - Effectiveness of spinal endoscopic adhesiolysis in post lumbar surgery syndrome: a systematic review. AB - BACKGROUND: Post lumbar surgery syndrome with persistent chronic low back and lower extremity pain is common in the United States. Epidural fibrosis may account for as much as 20% to 36% of all cases of failed back surgery syndrome (FBSS). Percutaneous adhesiolysis with a catheter or direct visualization of the spinal canal and the contents with an endoscope are techniques employed in resistant cases when patients fail to respond to conservative modalities of treatment, including fluoroscopically directed epidural injections. Some patients failing to respond to percutaneous adhesiolysis are candidates for spinal endoscopic adhesiolysis. However, literature evaluating the effectiveness of spinal endoscopic adhesiolysis is sparse and discussions continue about its effectiveness, utility, and complications. STUDY DESIGN: A systematic review of the available literature. OBJECTIVE: To evaluate the effectiveness and safety of spinal endoscopic adhesiolysis in the management of chronic low back and lower extremity pain in post surgical patients with chronic recalcitrant pain, non responsive to conservative modalities of management and fluoroscopically directed epidural injections. METHODS: A search of relevant resources (PubMed, EMBASE, and the Cochrane database) was accomplished and the resulting publications were examined based on the inclusion/exclusion criteria set forth. Randomized controlled trials and observational studies were included in the search. Two reviewers assessed the studies' methodologies and outcomes. Randomized clinical trials were assessed and scored based on the criteria established by the Cochrane methodological assessment criteria of randomized clinical trials and the observational studies were assessed and scored based on the Agency for Healthcare Research and Quality (AHRQ) criteria. Clinical relevance was evaluated utilizing Cochrane review criteria. Analysis was conducted using 5 levels of evidence, ranging from Level I to III, with 3 subcategories in Level II. OUTCOME MEASURES: The primary outcome measure was pain relief (> or = 50%) in follow-up for at least 6 months. Pain relief for longer than 6 months was considered long-term and 6 months or less was considered short-term. The secondary outcome measures were functional and psychological status, return to work, patient satisfaction, and opioid intake. RESULTS: Of the 13 studies considered for inclusion, one randomized trial and 5 observational studies met inclusion criteria for evidence synthesis based on the inclusion criteria and methodologic quality scores of 50 or more. The indicated level of evidence for endoscopic adhesiolysis is Level II 1 or II-2 evidence for short- and long-term relief based on the U.S. Preventive Services Task Force (USPSTF) criteria. LIMITATIONS: There was a paucity of literature for randomized trials. CONCLUSION: Spinal endoscopic adhesiolysis may be used as an effective treatment modality for chronic refractory low back pain and radiculopathy that is related to epidural adhesions. PMID- 19305489 TI - Systematic assessment of diagnostic accuracy and therapeutic utility of lumbar facet joint interventions. AB - BACKGROUND: Lumbar facet joints are a well recognized source of low back pain and referred pain in the lower extremity in patients with chronic low back pain. Conventional clinical features and other non-invasive diagnostic modalities are unreliable in diagnosing lumbar zygapophysial joint pain. Controlled diagnostic studies have shown the prevalence of lumbar facet joint pain in 27% to 40% of the patients with chronic low back pain without disc displacement or radiculitis, with a false-positive rate of 27% to 47% with a single diagnostic block. STUDY DESIGN: A systematic review of diagnostic and therapeutic lumbar facet joint interventions. OBJECTIVE: To determine the clinical utility of diagnostic and therapeutic lumbar facet joint interventions in managing chronic low back pain of facet joint origin. METHODS: Review of the literature for clinical studies on efficacy and utility of facet joint interventions in diagnosing and managing facet joint pain was performed according to the Agency for Healthcare Research and Quality (AHRQ) criteria for diagnostic studies and observational studies and the Cochrane Musculoskeletal Review Group criteria as utilized for interventional techniques for randomized trials. Data sources included relevant literature of the English language identified through searches of Medline and EMBASE from 1966 to December 2008 and manual searches of bibliographies of known primary and review articles. Analysis results were performed for diagnostic and therapeutic interventions separately. LEVEL OF EVIDENCE: The level of evidence was defined as Level I, II, or III with 3 subcategories in Level II based on the quality of evidence developed by the U.S. Preventive Services Task Force (USPSTF) for therapeutic interventions. OUTCOME MEASURES: For diagnostic interventions, studies must have been performed utilizing controlled local anesthetic blocks. Pain relief was categorized as at least 80% pain relief from baseline pain and ability to perform previously painful movements. For therapeutic interventions, the primary outcome measure was pain relief with secondary outcome measures of improvement in functional status, psychological status, return to work, and reduction in opioid intake. For therapeutic interventions, short-term pain relief was defined as relief lasting 6 months or less and long-term relief as longer than 6 months. RESULTS: Based on USPSTF criteria, evidence showed Level I or II-1 for diagnostic facet joint nerve blocks. Based on the review of included therapeutic studies, Level II-1 to II-2 evidence was indicated for lumbar facet joint nerve blocks with indicated level of evidence of Level II-2 to II-3 for lumbar radiofrequency neurotomy. LIMITATIONS: The shortcoming of this systematic review of lumbar facet joint interventions is the paucity of published literature. CONCLUSION: The evidence for diagnosis of lumbar facet joint pain with controlled local anesthetic blocks is Level I or II-1. The indicated level of evidence for therapeutic lumbar facet joint interventions is Level II-1 or II 2 for lumbar facet joint nerve blocks, Level II-2 or II-3 evidence for radiofrequency neurotomy, and Level III (limited) evidence for intraarticular injections. PMID- 19305490 TI - Systematic assessment of the quality of research studies of conventional and alternative treatment(s) of primary headache. AB - BACKGROUND: Diversity of treatments used for headache, and varied quality of research conduct and reporting make it difficult to accurately assess the literature and to determine the best treatment(s) for patients. OBJECTIVES: To compare the quality of available research evidence describing the effects and outcomes of conventional, and complementary and alternative medicine (CAM) approaches to treating primary (migraine, tension, and/or cluster-type) headache. STUDY DESIGN: A systematic review of quality of research studies of conventional and alternative treatment(s) of primary headache. METHODS: Randomized, controlled clinical trials (RCTs) of treatment(s) of chronic primary headache (in English between 1979 to June 2004) were searched through MEDLINE, PsycInfo, EMBASE, Cochrane Library, and the NIH databases. Studies were evaluated using standard approaches for assessing and analyzing quality indicators. RESULTS: 125 studies of conventional, and 121 CAM treatments met inclusion criteria. 80% of studies of conventional treatment(s) reported positive effects (p<0.05), versus 73% of studies of CAM approaches (chi(2) = 3.798, 1 df, p=0.051). Overall, the literature addressing the treatment of primary headache received a mean Jadad score of 2.72 out of 5 (SD 1.1). The mean Jadad score for studies of conventional therapeutics was significantly better than for those studies of CAM approaches: 3.21 +/- 0.9 vs 2.23 +/- 1.1 (t=7.72, 246 df, mean difference 0.98, p < 0.0005). CONCLUSIONS: Studies of conventional treatments scored higher on reporting quality than studies of CAM approaches. It is possible that these differences may reflect distinctions in 1) methodologic integrity, 2) therapeutic paradigm(s), and/or 3) bias(es) in the approach(es) used to evaluate certain types of therapies. Each of these possibilities -- and the implications -- is addressed and considered. PMID- 19305491 TI - Recent advances relating to the clinical application of naked monoclonal antibodies in solid tumors. AB - This review focuses on the recent advances in clinical data regarding antibody based therapy in the management of solid tumors. We also discuss perspectives on antibody-based therapy in the future. Thorough understanding of the complex interactions between components of the immunological response has led to interest in the concept of immune-mediated therapy for solid tumors. Over the last few years, several humanized and chimeric monoclonal antibodies (MAbs) targeting human epidermal receptor 2 (HER2), epidermal growth factor receptor (EGFR), and vascular endothelial growth factor (VEGF) have been employed in treating solid tumors, including breast, colorectal, lung, head and neck, and gynecologic cancers. Trastuzumab, bevacizumab, cetuximab, and panitumumab are MAbs that are most widely used in clinical practice with acceptable rates of adverse events. Combination of MAbs with small-molecule inhibitors of the same pathway could potentially increase the efficacy and specificity of antibody-based treatment. Immune-mediated effects may be further exploited with the use of bivalent molecules. PMID- 19305493 TI - Alveolar epithelial type II cells activate alveolar macrophages and mitigate P. Aeruginosa infection. AB - Although alveolar epithelial type II cells (AECII) perform substantial roles in the maintenance of alveolar integrity, the extent of their contributions to immune defense is poorly understood. Here, we demonstrate that AECII activates alveolar macrophages (AM) functions, such as phagocytosis using a conditioned medium from AECII infected by P. aeruginosa. AECII-derived chemokine MCP-1, a monocyte chemoattractant protein, was identified as a main factor in enhancing AM function. We proposed that the enhanced immune potency of AECII may play a critical role in alleviation of bacterial propagation and pneumonia. The ability of phagocytosis and superoxide release by AM was reduced by MCP-1 neutralizing antibodies. Furthermore, MCP-1(-/-) mice showed an increased bacterial burden under PAO1 and PAK infection vs. wt littermates. AM from MCP-1(-/-) mice also demonstrated less superoxide and impaired phagocytosis over the controls. In addition, AECII conditioned medium increased the host defense of airway in MCP-1( /-) mice through the activation of AM function. Mechanistically, we found that Lyn mediated NFkappaB activation led to increased gene expression and secretion of MCP-1. Consequently Lyn(-/-) mice had reduced MCP-1 secretion and resulted in a decrease in superoxide and phagocytosis by AM. Collectively, our data indicate that AECII may serve as an immune booster for fighting bacterial infections, particularly in severe immunocompromised conditions. PMID- 19305494 TI - Heparanase regulates levels of syndecan-1 in the nucleus. AB - Syndecan-1 is a transmembrane heparan sulfate-bearing proteoglycan known to regulate multiple biological functions at the cell surface and within the extracellular matrix. Its functional activity can be modulated by heparanase, an enzyme that cleaves heparan sulfate chains and whose expression has been associated with an aggressive phenotype in many cancers. In addition to remodeling syndecan-1 by cleaving its heparan sulfate chains, heparanase influences syndecan-1 location by upregulating expression of enzymes that accelerate its shedding from the cell surface. In the present study we discovered that heparanase also alters the level of nuclear syndecan-1. Upon upregulation of heparanase expression or following addition of recombinant heparanase to myeloma cells, the nuclear localization of syndecan-1 drops dramatically as revealed by confocal microscopy, western blotting and quantification by ELISA. This effect requires enzymatically active heparanase because cells expressing high levels of mutated, enzymatically inactive heparanase, failed to diminish syndecan-1 levels in the nucleus. Although heparan sulfate function within the nucleus is not well understood, there is emerging evidence that it may act to repress transcriptional activity. The resulting changes in gene expression facilitated by the loss of nuclear syndecan-1 could explain how heparanase enhances expression of MMP-9, VEGF, tissue factor and perhaps other effectors that condition the tumor microenvironment to promote an aggressive cancer phenotype. PMID- 19305495 TI - Enzymatic primer-extension with glycerol-nucleoside triphosphates on DNA templates. AB - BACKGROUND: Glycerol nucleic acid (GNA) has an acyclic phosphoglycerol backbone repeat-unit, but forms stable duplexes based on Watson-Crick base-pairing. Because of its structural simplicity, GNA is of particular interest with respect to the possibility of evolving functional polymers by in vitro selection. Template-dependent GNA synthesis is essential to any GNA-based selection system. PRINCIPAL FINDINGS: In this study, we investigated the ability of various DNA polymerases to use glycerol-nucleoside triphosphates (gNTPs) as substrates for GNA synthesis on DNA templates. Therminator DNA polymerase catalyzes quantitative primer-extension by the incorporation of two glyceronucleotides, with much less efficient extension up to five glyceronucleotides. Steady-state kinetic experiments suggested that GNA synthesis by Therminator was affected by both decreased catalytic rates and weakened substrate binding, especially for pyrimidines. In an attempt to improve pyrimidine incorporation by providing additional stacking interactions, we synthesized two new gNTP analogs with 5 propynyl substituted pyrimidine nucleobases. This led to more efficient incorporation of gC, but not gT. CONCLUSIONS: We suggest that directed evolution of Therminator might lead to mutants with improved substrate binding and catalytic efficiency. PMID- 19305496 TI - Age-related cellular copper dynamics in the fungal ageing model Podospora anserina and in ageing human fibroblasts. AB - In previous investigations an impact of cellular copper homeostasis on ageing of the ascomycete Podospora anserina has been demonstrated. Here we provide new data indicating that mitochondria play a major role in this process. Determination of copper in the cytosolic fraction using total reflection X-ray fluorescence spectroscopy analysis and eGfp reporter gene studies indicate an age-related increase of cytosolic copper levels. We show that components of the mitochondrial matrix (i.e. eGFP targeted to mitochondria) become released from the organelle during ageing. Decreasing the accessibility of mitochondrial copper in P. anserina via targeting a copper metallothionein to the mitochondrial matrix was found to result in a switch from a copper-dependent cytochrome-c oxidase to a copper-independent alternative oxidase type of respiration and results in lifespan extension. In addition, we demonstrate that increased copper concentrations in the culture medium lead to the appearance of senescence biomarkers in human diploid fibroblasts (HDFs). Significantly, expression of copper-regulated genes is induced during in vitro ageing in medium devoid of excess copper suggesting that cytosolic copper levels also increase during senescence of HDFs. These data suggest that the identified molecular pathway of age-dependent copper dynamics may not be restricted to P. anserina but may be conserved from lower eukaryotes to humans. PMID- 19305497 TI - Glucose amplifies fatty acid-induced endoplasmic reticulum stress in pancreatic beta-cells via activation of mTORC1. AB - BACKGROUND: Palmitate is a potent inducer of endoplasmic reticulum (ER) stress in beta-cells. In type 2 diabetes, glucose amplifies fatty-acid toxicity for pancreatic beta-cells, leading to beta-cell dysfunction and death. Why glucose exacerbates beta-cell lipotoxicity is largely unknown. Glucose stimulates mTORC1, an important nutrient sensor involved in the regulation of cellular stress. Our study tested the hypothesis that glucose augments lipotoxicity by stimulating mTORC1 leading to increased beta-cell ER stress. PRINCIPAL FINDINGS: We found that glucose amplifies palmitate-induced ER stress by increasing IRE1alpha protein levels and activating the JNK pathway, leading to increased beta-cell apoptosis. Moreover, glucose increased mTORC1 activity and its inhibition by rapamycin decreased beta-cell apoptosis under conditions of glucolipotoxicity. Inhibition of mTORC1 by rapamycin did not affect proinsulin and total protein synthesis in beta-cells incubated at high glucose with palmitate. However, it decreased IRE1alpha expression and signaling and inhibited JNK pathway activation. In TSC2-deficient mouse embryonic fibroblasts, in which mTORC1 is constitutively active, mTORC1 regulated the stimulation of JNK by ER stressors, but not in response to anisomycin, which activates JNK independent of ER stress. Finally, we found that JNK inhibition decreased beta-cell apoptosis under conditions of glucolipotoxicity. CONCLUSIONS/SIGNIFICANCE: Collectively, our findings suggest that mTORC1 mediates glucose amplification of lipotoxicity, acting through activation of ER stress and JNK. Thus, mTORC1 is an important transducer of ER stress in beta-cell glucolipotoxicity. Moreover, in stressed beta-cells mTORC1 inhibition decreases IRE1alpha protein expression and JNK activity without affecting ER protein load, suggesting that mTORC1 regulates the beta-cell stress response to glucose and fatty acids by modulating the synthesis and activity of specific proteins involved in the execution of the ER stress response. This novel paradigm may have important implications for understanding beta-cell failure in type 2 diabetes. PMID- 19305498 TI - The oncogenic EWS-FLI1 protein binds in vivo GGAA microsatellite sequences with potential transcriptional activation function. AB - The fusion between EWS and ETS family members is a key oncogenic event in Ewing tumors and important EWS-FLI1 target genes have been identified. However, until now, the search for EWS-FLI1 targets has been limited to promoter regions and no genome-wide comprehensive analysis of in vivo EWS-FLI1 binding sites has been undertaken. Using a ChIP-Seq approach to investigate EWS-FLI1-bound DNA sequences in two Ewing cell lines, we show that this chimeric transcription factor preferentially binds two types of sequences including consensus ETS motifs and microsatellite sequences. Most bound sites are found outside promoter regions. Microsatellites containing more than 9 GGAA repeats are very significantly enriched in EWS-FLI1 immunoprecipitates. Moreover, in reporter gene experiments, the transcription activation is highly dependent upon the number of repeats that are included in the construct. Importantly, in vivo EWS-FLI1-bound microsatellites are significantly associated with EWS-FLI1-driven gene activation. Put together, these results point out the likely contribution of microsatellite elements to long-distance transcription regulation and to oncogenesis. PMID- 19305499 TI - Ultrastructural proof of polyomavirus in Merkel cell carcinoma tumour cells and its absence in small cell carcinoma of the lung. AB - BACKGROUND: A new virus called the Merkel Cell Polyomavirus (MCPyV) has recently been found in Merkel Cell Carcinoma (MCC). MCC is a rare aggressive small cell neuroendocrine carcinoma primarily derived from the skin, morphologically indistinguishable from small cell lung carcinoma (SCLC). So far the actual presence of the virus in MCC tumour cells on a morphological level has not been demonstrated, and the presence of MCPyV in other small cell neuroendocrine carcinomas has not been studied yet. METHODOLOGY/PRINCIPAL FINDINGS: We investigated MCC tissue samples from five patients and SCLCs from ten patients for the presence of MCPyV-DNA by PCR and sequencing. Electron microscopy was used to search ultrastructurally for morphological presence of the virus in MCPyV-DNA positive samples. MCPyV was detected in two out of five primary MCCs. In one MCC patient MCPyV-DNA was detected in the primary tumour as well as in the metastasis, strongly suggesting integration of MCPyV in the cellular DNA of the tumour in this patient. In the primary MCC of another patient viral particles in tumour cell nuclei and cytoplasm were identified by electron microscopy, indicating active viral replication in the tumour cells. In none of the SCLCs MCPyV-DNA was detected. CONCLUSIONS/SIGNIFICANCE: Our results strongly suggest that MCPyV is an oncogenic polyomavirus in humans, and is potentially causally related to the development of MCC but not to the morphological similar SCLC. PMID- 19305500 TI - Mutational meltdown in primary endosymbionts: selection limits Muller's ratchet. AB - BACKGROUND: Primary bacterial endosymbionts of insects (p-endosymbionts) are thought to be undergoing the process of Muller's ratchet where they accrue slightly deleterious mutations due to genetic drift in small populations with negligible recombination rates. If this process were to go unchecked over time, theory predicts mutational meltdown and eventual extinction. Although genome degradation is common among p-endosymbionts, we do not observe widespread p endosymbiont extinction, suggesting that Muller's ratchet may be slowed or even stopped over time. For example, selection may act to slow the effects of Muller's ratchet by removing slightly deleterious mutations before they go to fixation thereby causing a decrease in nucleotide substitutions rates in older p endosymbiont lineages. METHODOLOGY/PRINCIPAL FINDINGS: To determine whether selection is slowing the effects of Muller's ratchet, we determined the age of the Candidatus Riesia/sucking louse assemblage and analyzed the nucleotide substitution rates of several p-endosymbiont lineages that differ in the length of time that they have been associated with their insect hosts. We find that Riesia is the youngest p-endosymbiont known to date, and has been associated with its louse hosts for only 13-25 My. Further, it is the fastest evolving p endosymbiont with substitution rates of 19-34% per 50 My. When comparing Riesia to other insect p-endosymbionts, we find that nucleotide substitution rates decrease dramatically as the age of endosymbiosis increases. CONCLUSIONS/SIGNIFICANCE: A decrease in nucleotide substitution rates over time suggests that selection may be limiting the effects of Muller's ratchet by removing individuals with the highest mutational loads and decreasing the rate at which new mutations become fixed. This countering effect of selection could slow the overall rate of endosymbiont extinction. PMID- 19305501 TI - Psychological typhoon eye in the 2008 Wenchuan earthquake. AB - BACKGROUND: On May 12, 2008, an earthquake measuring 8.0 on the Richter scale jolted Wenchuan, China, leading to 69,227 deaths and 374,643 injured, with 17,923 listed as missing as of Sept. 25, 2008, and shook the whole nation. We assessed the devastating effects on people's post-earthquake concern about safety and health. METHODOLOGY/PRINCIPAL FINDINGS: From June 4 to July 15, 2008, we surveyed a convenience sample of 2,262 adults on their post-earthquake concern about safety and health. Residents in non-devastated areas (Fujian and Hunan Provinces, and Beijing) and devastated areas (Sichuan and Gansu Provinces) responded to a questionnaire of 5 questions regarding safety measures, epidemic disease, medical workers, psychological workers, and medication. The ANOVAs showed a significant effect of residential devastation level on the estimated number of safety measures needed, the estimated probability of the outbreak of an epidemic, and the estimated number of medical and psychological workers needed (Ps<0.001). The post-earthquake concern decreased significantly as the level of residential devastation increased. Because of the similarity with the meteorological phenomenon of the eye of a typhoon, we dubbed these findings a "Psychological Typhoon Eye": the closer to the center of the devastated areas, the less the concern about safety and health a resident felt. CONCLUSIONS/SIGNIFICANCE: Contrary to common perception and ripple effect that the impact of an unfortunate event decays gradually as ripples spread outward from a center, a "Psychological Typhoon Eye" effect was observed where the post-earthquake concern was at its lowest level in the extremely devastated areas. The resultant findings may have implications for Chinese governmental strategies for putting "psychological comfort" into effect. PMID- 19305502 TI - Phagocytic superoxide specifically damages an extracytoplasmic target to inhibit or kill Salmonella. AB - BACKGROUND: The phagocytic oxidative burst is a primary effector of innate immunity that protects against bacterial infection. However, the mechanism by which reactive oxygen species (ROS) kill or inhibit bacteria is not known. It is often assumed that DNA is a primary target of oxidative damage, consistent with known effects of endogenously produced ROS in the bacterial cytoplasm. But most studies fail to distinguish between effects of host derived ROS versus damage caused by endogenous bacterial sources. We took advantage of both the ability of Salmonella enterica serovar Typhimurium to survive in macrophages and the genetic tractability of the system to test the hypothesis that phagocytic superoxide damages cytoplasmic targets including DNA. METHODOLOGY/PRINCIPAL FINDINGS: SodCI is a periplasmic Cu-Zn superoxide dismutase (SOD) that contributes to the survival of Salmonella Typhimurium in macrophages. Through competitive virulence assays, we asked if sodCI has a genetic interaction with various cytoplasmic systems. We found that SodCI acts independently of cytoplasmic SODs, SodA and SodB. In addition, SodCI acts independently of the base excision repair system and RuvAB, involved in DNA repair. Although sodCI did show genetic interaction with recA, this was apparently independent of recombination and is presumably due to the pleiotropic effects of a recA mutation. CONCLUSIONS/SIGNIFICANCE: Taken together, these results suggest that bacterial inhibition by phagocytic superoxide is primarily the result of damage to an extracytoplasmic target. PMID- 19305503 TI - Comparison of gene expression profile in embryonic mesencephalon and neuronal primary cultures. AB - In the mammalian central nervous system (CNS) an important contingent of dopaminergic neurons are localized in the substantia nigra and in the ventral tegmental area of the ventral midbrain. They constitute an anatomically and functionally heterogeneous group of cells involved in a variety of regulatory mechanisms, from locomotion to emotional/motivational behavior. Midbrain dopaminergic neuron (mDA) primary cultures represent a useful tool to study molecular mechanisms involved in their development and maintenance. Considerable information has been gathered on the mDA neurons development and maturation in vivo, as well as on the molecular features of mDA primary cultures. Here we investigated in detail the gene expression differences between the tissue of origin and ventral midbrain primary cultures enriched in mDA neurons, using microarray technique. We integrated the results based on different re-annotations of the microarray probes. By using knowledge-based gene network techniques and promoter sequence analysis, we also uncovered mechanisms that might regulate the expression of CNS genes involved in the definition of the identity of specific cell types in the ventral midbrain. We integrate bioinformatics and functional genomics, together with developmental neurobiology. Moreover, we propose guidelines for the computational analysis of microarray gene expression data. Our findings help to clarify some molecular aspects of the development and differentiation of DA neurons within the midbrain. PMID- 19305504 TI - A cross-study transcriptional analysis of Parkinson's disease. AB - The study of Parkinson's disease (PD), like other complex neurodegenerative disorders, is limited by access to brain tissue from patients with a confirmed diagnosis. Alternatively the study of peripheral tissues may offer some insight into the molecular basis of disease susceptibility and progression, but this approach still relies on brain tissue to benchmark relevant molecular changes against. Several studies have reported whole-genome expression profiling in post mortem brain but reported concordance between these analyses is lacking. Here we apply a standardised pathway analysis to seven independent case-control studies, and demonstrate increased concordance between data sets. Moreover data convergence increased when the analysis was limited to the five substantia nigra (SN) data sets; this highlighted the down regulation of dopamine receptor signaling and insulin-like growth factor 1 (IGF1) signaling pathways. We also show that case-control comparisons of affected post mortem brain tissue are more likely to reflect terminal cytoarchitectural differences rather than primary pathogenic mechanisms. The implementation of a correction factor for dopaminergic neuronal loss predictably resulted in the loss of significance of the dopamine signaling pathway while axon guidance pathways increased in significance. Interestingly the IGF1 signaling pathway was also over-represented when data from non-SN areas, unaffected or only terminally affected in PD, were considered. Our findings suggest that there is greater concordance in PD whole-genome expression profiling when standardised pathway membership rather than ranked gene list is used for comparison. PMID- 19305505 TI - A fast Na+/Ca2+-based action potential in a marine diatom. AB - BACKGROUND: Electrical impulses in animals play essential roles in co-ordinating an array of physiological functions including movement, secretion, environmental sensing and development. Underpinning many of these electrical signals is a fast Na+-based action potential that has been fully characterised only in cells associated with the neuromuscular systems of multicellular animals. Such rapid action potentials are thought to have evolved with the first metazoans, with cnidarians being the earliest representatives. The present study demonstrates that a unicellular protist, the marine diatom Odontella sinensis, can also generate a fast Na+/Ca2+ based action potential that has remarkably similar biophysical and pharmacological properties to invertebrates and vertebrate cardiac and skeletal muscle cells. METHODOLOGY/PRINCIPAL FINDINGS: The kinetic, ionic and pharmacological properties of the rapid diatom action potential were examined using single electrode current and voltage clamp techniques. Overall, the characteristics of the fast diatom currents most closely resemble those of vertebrate and invertebrate muscle Na+/Ca2+ currents. CONCLUSIONS/SIGNIFICANCE: This is the first demonstration of voltage-activated Na+ channels and the capacity to generate fast Na+-based action potentials in a unicellular photosynthetic organism. The biophysical and pharmacological characteristics together with the presence of a voltage activated Na+/Ca2+ channel homologue in the recently sequenced genome of the diatom Thalassiosira pseudonana, provides direct evidence supporting the hypothesis that this rapid signalling mechanism arose in ancestral unicellular eukaryotes and has been retained in at least two phylogenetically distant lineages of eukaryotes; opisthokonts and the stramenopiles. The functional role of the fast animal-like action potential in diatoms remains to be elucidated but is likely involved in rapid environmental sensing of these widespread and successful marine protists. PMID- 19305506 TI - Filling kinetic gaps: dynamic modeling of metabolism where detailed kinetic information is lacking. AB - BACKGROUND: Integrative analysis between dynamical modeling of metabolic networks and data obtained from high throughput technology represents a worthy effort toward a holistic understanding of the link among phenotype and dynamical response. Even though the theoretical foundation for modeling metabolic network has been extensively treated elsewhere, the lack of kinetic information has limited the analysis in most of the cases. To overcome this constraint, we present and illustrate a new statistical approach that has two purposes: integrate high throughput data and survey the general dynamical mechanisms emerging for a slightly perturbed metabolic network. METHODOLOGY/PRINCIPAL FINDINGS: This paper presents a statistic framework capable to study how and how fast the metabolites participating in a perturbed metabolic network reach a steady-state. Instead of requiring accurate kinetic information, this approach uses high throughput metabolome technology to define a feasible kinetic library, which constitutes the base for identifying, statistical and dynamical properties during the relaxation. For the sake of illustration we have applied this approach to the human Red blood cell metabolism (hRBC) and its capacity to predict temporal phenomena was evaluated. Remarkable, the main dynamical properties obtained from a detailed kinetic model in hRBC were recovered by our statistical approach. Furthermore, robust properties in time scale and metabolite organization were identify and one concluded that they are a consequence of the combined performance of redundancies and variability in metabolite participation. CONCLUSIONS/SIGNIFICANCE: In this work we present an approach that integrates high throughput metabolome data to define the dynamic behavior of a slightly perturbed metabolic network where kinetic information is lacking. Having information of metabolite concentrations at steady-state, this method has significant relevance due its potential scope to analyze others genome scale metabolic reconstructions. Thus, I expect this approach will significantly contribute to explore the relationship between dynamic and physiology in other metabolic reconstructions, particularly those whose kinetic information is practically nulls. For instances, I envisage that this approach can be useful in genomic medicine or pharmacogenomics, where the estimation of time scales and the identification of metabolite organization may be crucial to characterize and identify (dis)functional stages. PMID- 19305507 TI - Coordinated activation of candidate proto-oncogenes and cancer testes antigens via promoter demethylation in head and neck cancer and lung cancer. AB - BACKGROUND: Epigenetic alterations have been implicated in the pathogenesis of solid tumors, however, proto-oncogenes activated by promoter demethylation have been sporadically reported. We used an integrative method to analyze expression in primary head and neck squamous cell carcinoma (HNSCC) and pharmacologically demethylated cell lines to identify aberrantly demethylated and expressed candidate proto-oncogenes and cancer testes antigens in HNSCC. METHODOLOGY/PRINCIPAL FINDINGS: We noted coordinated promoter demethylation and simultaneous transcriptional upregulation of proto-oncogene candidates with promoter homology, and phylogenetic footprinting of these promoters demonstrated potential recognition sites for the transcription factor BORIS. Aberrant BORIS expression correlated with upregulation of candidate proto-oncogenes in multiple human malignancies including primary non-small cell lung cancers and HNSCC, induced coordinated proto-oncogene specific promoter demethylation and expression in non-tumorigenic cells, and transformed NIH3T3 cells. CONCLUSIONS/SIGNIFICANCE: Coordinated, epigenetic unmasking of multiple genes with growth promoting activity occurs in aerodigestive cancers, and BORIS is implicated in the coordinated promoter demethylation and reactivation of epigenetically silenced genes in human cancers. PMID- 19305508 TI - Reducing glycosphingolipid content in adipose tissue of obese mice restores insulin sensitivity, adipogenesis and reduces inflammation. AB - Adipose tissue is a critical mediator in obesity-induced insulin resistance. Previously we have demonstrated that pharmacological lowering of glycosphingolipids and subsequently GM3 by using the iminosugar AMP-DNM, strikingly improves glycemic control. Here we studied the effects of AMP-DNM on adipose tissue function and inflammation in detail to provide an explanation for the observed improved glucose homeostasis. Leptin-deficient obese (Lep(Ob)) mice were fed AMP-DNM and its effects on insulin signalling, adipogenesis and inflammation were monitored in fat tissue. We show that reduction of glycosphingolipid biosynthesis in adipose tissue of Lep(Ob) mice restores insulin signalling in isolated ex vivo insulin-stimulated adipocytes. We observed improved adipogenesis as the number of larger adipocytes was reduced and expression of genes like peroxisome proliferator-activated receptor (PPAR) gamma, insulin responsive glucose transporter (GLUT)-4 and adipsin increased. In addition, we found that adiponectin gene expression and protein were increased by AMP-DNM. As a consequence of this improved function of fat tissue we observed less inflammation, which was characterized by reduced numbers of adipose tissue macrophages (crown-like structures) and reduced levels of the macrophage chemo attractants monocyte-chemoattractant protein-1 (Mcp-1/Ccl2) and osteopontin (OPN). In conclusion, pharmacological lowering of glycosphingolipids by inhibition of glucosylceramide biosynthesis improves adipocyte function and as a consequence reduces inflammation in adipose tissue of obese animals. PMID- 19305509 TI - A specialized odor memory buffer in primary olfactory cortex. AB - BACKGROUND: The neural substrates of olfactory working memory are unknown. We addressed the questions of whether olfactory working memory involves a verbal representation of the odor, or a sensory image of the odor, or both, and the location of the neural substrates of these processes. METHODOLOGY/PRINCIPAL FINDINGS: We used functional magnetic resonance imaging to measure activity in the brains of subjects who were remembering either nameable or unnameable odorants. We found a double dissociation whereby remembering nameable odorants was reflected in sustained activity in prefrontal language areas, and remembering unnameable odorants was reflected in sustained activity in primary olfactory cortex. CONCLUSIONS/SIGNIFICANCE: These findings suggest a novel dedicated mechanism in primary olfactory cortex, where odor information is maintained in temporary storage to subserve ongoing tasks. PMID- 19305510 TI - Phase shifting capacity of the circadian pacemaker determined by the SCN neuronal network organization. AB - BACKGROUND: In mammals, a major circadian pacemaker that drives daily rhythms is located in the suprachiasmatic nuclei (SCN), at the base of the hypothalamus. The SCN receive direct light input via the retino-hypothalamic tract. Light during the early night induces phase delays of circadian rhythms while during the late night it leads to phase advances. The effects of light on the circadian system are strongly dependent on the photoperiod to which animals are exposed. An explanation for this phenomenon is currently lacking. METHODOLOGY AND PRINCIPAL FINDINGS: We recorded running wheel activity in C57 mice and observed large amplitude phase shifts in short photoperiods and small shifts in long photoperiods. We investigated whether these different light responses under short and long days are expressed within the SCN by electrophysiological recordings of electrical impulse frequency in SCN slices. Application of N-methyl-D-aspartate (NMDA) induced sustained increments in electrical activity that were not significantly different in the slices from long and short photoperiods. These responses led to large phase shifts in slices from short days and small phase shifts in slices from long days. An analysis of neuronal subpopulation activity revealed that in short days the amplitude of the rhythm was larger than in long days. CONCLUSIONS: The data indicate that the photoperiodic dependent phase responses are intrinsic to the SCN. In contrast to earlier predictions from limit cycle theory, we observed large phase shifting responses in high amplitude rhythms in slices from short days, and small shifts in low amplitude rhythms in slices from long days. We conclude that the photoperiodic dependent phase responses are determined by the SCN and propose that synchronization among SCN neurons enhances the phase shifting capacity of the circadian system. PMID- 19305511 TI - Sperm from hyh mice carrying a point mutation in alphaSNAP have a defect in acrosome reaction. AB - Hydrocephalus with hop gait (hyh) is a recessive inheritable disease that arose spontaneously in a mouse strain. A missense mutation in the Napa gene that results in the substitution of a methionine for isoleucine at position 105 (M105I) of alphaSNAP has been detected in these animals. alphaSNAP is a ubiquitous protein that plays a key role in membrane fusion and exocytosis. In this study, we found that male hyh mice with a mild phenotype produced morphologically normal and motile sperm, but had a strongly reduced fertility. When stimulated with progesterone or A23187 (a calcium ionophore), sperm from these animals had a defective acrosome reaction. It has been reported that the M105I mutation affects the expression but not the function of the protein. Consistent with an hypomorphic phenotype, the testes and epididymides of hyh mice had low amounts of the mutated protein. In contrast, sperm had alphaSNAP levels indistinguishable from those found in wild type cells, suggesting that the mutated protein is not fully functional for acrosomal exocytosis. Corroborating this possibility, addition of recombinant wild type alphaSNAP rescued exocytosis in streptolysin O-permeabilized sperm, while the mutant protein was ineffective. Moreover, addition of recombinant alphaSNAP. M105I inhibited acrosomal exocytosis in permeabilized human and wild type mouse sperm. We conclude that the M105I mutation affects the expression and also the function of alphaSNAP, and that a fully functional alphaSNAP is necessary for acrosomal exocytosis, a key event in fertilization. PMID- 19305512 TI - A VARIATIONAL FRAMEWORK FOR PARTIALLY OCCLUDED IMAGE SEGMENTATION USING COARSE TO FINE SHAPE ALIGNMENT AND SEMI-PARAMETRIC DENSITY APPROXIMATION. AB - In this paper, we propose a variational framework which combines top-down and bottom-up information to address the challenge of partially occluded image segmentation. The algorithm applies shape priors and divides shape learning into shape mode clustering and non-rigid transformation estimation to handle intraclass and interclass coarse to fine variations. A semi-parametric density approximation using adaptive meanshift and L(2)E robust estimation is used to model the likelihood. A set of real images is used to show the good performance of the algorithm. PMID- 19305513 TI - Analysis of Smoking Cessation Patterns Using a Stochastic Mixed-Effects Model With a Latent Cured State. AB - We develop a mixed model to capture the complex stochastic nature of tobacco abuse and dependence. This model describes transition processes among addiction and nonaddiction stages. An important innovation of our model is allowing an unobserved cure state, or permanent quitting, in contrast to transient quitting. This distinction is necessary to model data from situations where censoring prevents unambiguous determination that a person has been "cured." Moreover, the processes that describe transient and permanent quitting are likely to be different and have different policy-making implications. For example, when analyzing factors that influence smoking and can be targeted by interventions, it is more important to target those factors that are associated with permanent quitting rather than transient quitting.We apply our methodology to the Alpha Tocopherol, Beta-Carotene Cancer Prevention (ATBC) study, a large (29,133 participants) longitudinal cohort study. While ATBC was designed as a cancer prevention study, it contains unique information about the smoking status of each participant during every 4-month period of the study. These data are used to model smoking cessation patterns using a discrete-time stochastic mixed-effects model with three states: smoking, transient cessation, and permanent cessation (absorbent state). Random participant-specific transition probabilities among these states are used to account for participant-to-participant heterogeneity. Another important innovation in our article is to design computationally practical methods for dealing with the size of the dataset and complexity of the models. This is achieved using the marginal likelihood obtained by integrating over the Beta distribution of random effects. PMID- 19305514 TI - ENANTIOMERIC SYNTHESIS OF 2-C-METHYL-D-ERYTHRITOL 2, 4-CYCLODIPHOSPHATE. AB - Enantiomerically pure 2-C-methyl-D-erythritol 2,4-cyclodiphosphate 1 (ME-CPP) is synthesized from 1,2-O-isopropylidene-alpha-D-xylofuranose with facile phosphorylation in good yield. Subsequently, the synthesized enantiomerically pure 1 can be used as a substrate in IspG assays to identify inhibitors that may be developed into antibacterial drug leads. PMID- 19305515 TI - Homing in on an alga's threat-and therapeutic promise. PMID- 19305516 TI - State-Designated Special Needs, Post-Adoption Support, and State Fiscal Stress. AB - The Adoption and Safe Families Act of 2007 and its 2003 reauthorization offered bonuses to states that provide more children waiting in foster care with permanent families through adoption. Under ASFA, a majority of states increased the generosity of post-adoption financial support. Moreover, states increased the number and proportion of adopted children that received adoption assistance because the child had a special need. Administrative data indicate that states more diligently recorded the special needs of children, which was necessary to support increases in state requests for federal reimbursements and performance bonuses. PMID- 19305517 TI - HEALTH, HEALTH CARE, AND ECONOMIC DEVELOPMENT: Wealth, Health, and Health Services in Rural Rajasthan. PMID- 19305520 TI - Peroxidase Isozymes from Meloidogyne spp. and Their Origin. AB - Two peroxidase isozymes (Ef 0.43 and 0.53) were detected by electrophoretic analysis in homogenates of Meloidogyne arenaria, M. hapla, M. javanica, and M. incognita females reared on tomato. No peroxidase isozymes were detected electrophoretically in homogenates of adult males, preparasitic larvae, or eggs. Peroxidase isozymes from females reared on bean, eggplant, or tobacco differed from those from females reared on tomato. Bean and eggplant populations had a single peroxidase isozyme each, respectively Ef 0.21 and 0.28. No peroxidase isozymes were detected in tobacco populations under the conditions used, although total activity assays did reveal low levels of peroxidase activity in homogenates of tobacco populations. The peroxidase isozymes detected in females reared on tomato or bean appear similar to the peroxidase isozymes present in root-knot galls, adjacent ungalled roots, and roots from uninoculated plants of the corresponding hosts. The probability is discussed that most of the peroxittase activity associated with Meloidogyne spp. females is of host origin. PMID- 19305519 TI - Postnatal maturational properties of rat parafascicular thalamic neurons recorded in vitro. AB - Thalamic relay neurons have homogeneous, adult-like firing properties and similar morphology by 12 days postnatally (PN 12). Parafascicular (Pf) neurons have a different morphology compared with typical thalamic relay neurons, but the development of their electrophysiological properties is not well studied. Intracellular recordings in PN 12-50 Pf neurons revealed several heterogeneous firing patterns different from those in thalamic relay neurons. Two types of cells were identified: Type I cells displayed a fast afterhyperpolarization (AHP) followed by a large-amplitude, slow AHP; whereas Type II cells had only a fast AHP. These cell types had overlapping membrane properties but differences in excitability. Some properties of Pf neurons were adult-like by PN 12, but, unlike thalamic relay neurons, there were significant maturational changes thereafter, including decreased action potential (AP) duration, increased fast AHP amplitude and increased excitability. Pf neurons did not exhibit rhythmic bursting and generally lacked low-threshold spike (LTS) responses that characterize thalamic relay neurons. Pf neurons exhibited nonlinear I-V relationships, and only a third of the cells expressed the time and voltage-dependent hyperpolarization activated (Ih) current, which declined with age. These results indicate that the morphological differences between Pf neurons and typical thalamic relay neurons are paralleled by electrophysiological differences, and that Pf membrane properties change during postnatal development. PMID- 19305521 TI - Morphological Comparison of Second-Stage Juveniles of Six Populations of Meloidogyne hapla by SEM. AB - External morphology of second-stage juveniles of six populations of Meloidogyne hapla, hclonging to two cytological races (A and B), and one population each of M. arenaria, M. incognita, and M. javanica was compared by scanning electron microscopy (SEM). Race A of M. hapla included three facultatively parthenogenetic populations with haploid chromosome numbers of 15. 16, and 17; race B consisted of three mitotically parthenogenetic populations with somalic chromosome numhers of 45, 45, and 48. The mitotically parthenogenetic populations of M. arenaria, M. incognita, and M. javanica had 54, 41-43, and 44 chromosomes, respectively. Observations were made on head structures, lateral field, excretory pore, anal opening, and tail. Head morphology, including shape and proportion of labial disc and lips, expression of labial and cephalic sensilla, and markings on head region, was distinctly different for each species. M. hapla populations of race A were distinct from each other but showed much intrapopulatiou variation in head morphology. Populations of race B were different from those of race A and were very stable and quite similar in head morphology. Considerable inter- and intrapopulatiou variation made the structure of the lateral field, excretory pore, anal opening, and tail of little value in distinguishing species or populations. The results are discussed in relation to earlier SEM observations on the genus Helerodera. PMID- 19305522 TI - Gall Formation on Cirsium arvense by Ditylenchus dipsaci. AB - Ditylenchus dipsaci was found to cause gall formation on the stems of Cirsium arvense. The galls were characterized by extensive hypertrophy and hyperplasia, differentiation of nutritive tissue, nuclear modification, and a central cavity containing nematodes. These findings emphasize the importance of host response in investigations of host-parasite interactions and suggest that D. dipsaci may be evolving a host race by reproductive isolation within the confines of a plant gall. PMID- 19305523 TI - The Effects of Temperature on Pratylenchus scribneri and P. alleni Populations on Soybeans and Tomatoes. AB - In soil temperature tests, rates of Pratylenchus scribneri and P. alleni reproduction were measured at various lemperatures on 'Clark 63' and 'Cutler 71' soybeans and 'Rutgers' tomatoes. Recovered P. scribneri equaled or exceeded initial inoculum levels at temperatures of 27.5 C or higher on soybeans, and at 20 C or higher on tomatoes. Population increases were greatest at 3.5 C on both hosts. Populations increased on soybeans, but not on tomatoes, when soil temperature was raised from 25 to 35 C for either 3 or 9 days. Recovered P. alleni were less than the initial inoculum at 27.5 C but higher at 32 and 37.5 C and at 27.5 C on tomatoes, the lowest temperature tested for this nematode. In the field, soil temperatures 10 cm deep in eastern Kansas soybean growing areas reach 27.5 C only occasionally and for relatively short periods, which probably explains the relatively low and variable populations of P. scribneri and P. alleni on soybeans there. PMID- 19305524 TI - Relative Tolerance of Selected Soybean Cultivars to Hoplolaimus columbus and Possible Effects of Soil Temperature. AB - Eleven soybean [Glycine max (L.) Merr.] cultivars resistant to one or more plant parasitic nematodes, and one resistant to the Mexican bean beetle (Epilachna varivestis Muls.), were tested for susceptibility to Hoplolaimus columbus. All cultivars were parasitized, but nematode reproduction varied. 'Pickett-71' was the most susceptible host among the cultivars tested. 'Dyer' and three 'P.I. cultivars' were most tolerant when yield /plant and total yield were compared for fumigated and unfumigated plots, even though their yield potential was low. 'Hardee,' 'Coker 4504,' 'W-4,' 'D71-9257,' and 'ED-371' appeared tolerant throughout the growing season and yielded well in unfumigated soil. Infection and reproduction of H. columbus in 'Forrest' soybean roots were greater at 30 +/- 1 C than at 20 or 25 +/- 1 C. Plant height and root weight varied with the soil treatments. PMID- 19305525 TI - Effects of DCPA, EPTC, and Chlorpropham on Pathogenicity of Meloidogyne hapla to Alfalfa. AB - Treatments wilh the herbicides chlorpropham (isopropyl m-chlorocarbinilate), DCPA (dimethyl tetrachloroterephthalate), and El'TC (S-ethyl dipropylthiocarbamate), alone or in combination with Meloidogyne hapla Chilwood, significantly reduced the growth of both nemalode-resistant 'Nev Syn XX' atttt susceptible 'Ranger" alfalfa (Medicago saliva L.) seedlings. M. hapla infection of both alfalfas was reduced by all herbicides because of fewer available infective courts in the treated plants. EPTC, however, reduced resistance to M. hapla, as indicated by increased galling of 'Nev Syn XX' plants. PMID- 19305526 TI - Control of Meloidogyne javanica and M. arenaria on kenaf and roselle with genetic resistance and nematicides. AB - Kenaf (Hibiscus cannabinus) and roselle (H. sabdarifla) were evaluated in nematicide-treated and untreated field soil naturally infested with either Meloidogyne javanica or M. arenaria. Root-knot indices indicated that the kenaf breeding line j-l-113 had moderate resistance to M. javanica and low resistance to M. arenaria. Kenaf cv Everglades 71 was highly susceptible to both M. javanica and M. arenaria, and roselle breeding line A59-56 was highly resistant. Both nematode species reproduced on all plant entries, but more larvae were recovered from the soil in plots planted to Everglades 71 than in plots planted to j-l-l13 or A59-56. In untreated soil infested with M. javanica, dry-matter yields were greater (P = 0.05) for j-l-l13 and A59-56 than for Everglades 71. The percentages of live plants at harvest were: j-l-l13, 88; A59-56, 93; and Everglades 71, 9. Ethylene dibromide (1,2-dibromoethane) at 73.9 kg a.i./ha and DBCP (1,2-dibromo-3 chloropropane) at 17.6 kg a.i./ha increased dry-matter yields significantly for all entries planted in soil infested with M. arenaria. Carbofuran (2.3-dihydro 2,2-dimethyl-7-benzofuranyl methylcarbamate) at 5.9 kg a.i./ha did not increase the dry-matter yields of any entry. None of the nematicides increased the growth of any entry significantly in soil infested with M. javanica. PMID- 19305527 TI - Residues of Aldicarb and its Oxides in Beta vulgaris L. and Systemic Control of Heterodera schachtii. AB - Altlicarb residues in foliage of Beta vulgaris L. 21 days after transplanting to soil treated with 1-5 mug aldicarb/g soil were proportional to residues in storage roots, but 20 times as great. Initial concentrations of residues in roots 21 days after treatment were proportional to applied rates but declined by 56% when roots were stored 25 days at 24 C. Mean respective concentrations of aldicarb, aldicarb sulfoxide, and aldicarb sulfone were 8.7, 81.6, and 9.8% of the total residues. In separate tests, equivalent concentrations of toxic carbamates in roots resulted in similar levels of control of Heterodera schachtii. Systemic levels that completely suppressed development of females and males on sectioned roots were respectively 0.35 and 0.8 mug/g of root tissue. PMID- 19305528 TI - Meloidogyne megatyla n. sp. a Root-Knot Nematode from Loblolly Pine. AB - Meloidogyne megatyla n. sp. is described from Pinus taeda in North Carolina. Stylet knobs are distinctively high in proportion to width, giving an especially massive appearance to the knobs of larvae and males. Mean larval length is 416 mum and stylet length is 14.6 mum. The perineal pattern is composed of smooth striae, with a high arch, and is often somewhat rectangular. The relationship of M. megatyla to other Meloidogyne species is unclear, although a comparison is made with Meloidogyne incognita and Meloidogyne mali. Galling was slight; only about 50 eggs were produced per egg mass, and under greenhouse conditions a single generation may take more than 10 weeks. Meloidogyne megatyla n. sp. did not reproduce on any of the differential hosts commonly used to distinguish among Meloidogyne species. PMID- 19305529 TI - Helicotylenchus oleae n. sp. and H. neopaxilli n. sp. (Hoplolaimidae), Two New Spiral Nematodes Parasitic on Olive Trees in Italy. AB - Helicotylenchus oleae n. sp. and H. neopaxilli n. sp., from olive roots and soil in Italy, are described and illustrated. Helicotylenchus oleae can be distinguished from the related species H. canadensis and H. tunisiensis especially by the smaller styler, its distinctive tail shape, and a tail longer than one anal body width. Helicotylenchus neopaxilli differs from the close species H. paxilli by having a conical, anteriorly truncated labial region, shorter stylet, and phasmids always anterior to level of anus. Also illustrated and discussed are histopathological changes within feeder roots of olive caused by the feeding activity of the semi-endoparasitic H. oleae. PMID- 19305530 TI - The Influence of Temperature on Meloidogyne incognita on Soybean. AB - The effects of temperature and initial inoculum density of Meloidogyne incognita on soybean growth and nematode reproduction were investigated in greenhouse temperature tanks and in controlled-growth chambers. The interactions of initial inoculum density (P(i)) and soil temperature in effects on shoot growth were adequately described by multiple-regression models. At the highest temperatures (30 or 32/28 C), moderate to high inoculum killed many plants. A P(i) of 27,000 eggs/15-cm-diam pot retarded shoot growth at 26 C. Only the greatest P(i) (81,000 eggs/15-cm pot) suppressed shoot growth at 18, 22, or 20/16 C. Inoculation with 3,000 or 9,000 eggs/plant resulted in heavier root systems at all temperatures except 30 C. At that temperature, 9,000 eggs suppressed root growth. At 18 and 26 C, a Pi of 81,000 eggs was required to retard root growth. Nematode reproduction was related directly to temperature and P(i) except at a density of 81,000 eggs/15-cm pot. PMID- 19305531 TI - Proleptonchoides southindiae n. gen., n. sp., a New Leptonchoid from South India. AB - Proleptonchoides southindiae n. gen., n. sp. (Dorylaimida: Leptonchidae), is described from soil around false tobacco (Lobelia excelsa) and cardamom (Elettaria cardamomurn) in South India. P. southindiae is prodelphic, has a short constricted esophageal bulb and flanged odontophore, and is phylogenetically close to Proleptonchus. PMID- 19305532 TI - Behavioral Response of Nothanguina phyllobia to Selected Plant Species. AB - The silver-leaf nightshade nenmtode, Nothanguina phyllobia, is a promising biological control agent for its only reported host, Solanum elaeagnifolium Cav. When infective larvae of N. phyllobia and stem tissue of 39 econmnically important plant species were suspended in 0.5% water agar, nematodes aggregated about S. elaeagnifolium, Solanum carolinense L., Solanum melongena L., Solanum tuberosum L., and Prunus caroliniana (Mill.) Ait. Nematodes responded to Solanum spp. via positive chemotaxis and/or klinokinesis, but aggregated near tissue of P. caroliniana as a result of orthokinetic effects. Nematodes aggregated away from tissue of Hibiscus esculentus L., Triticum aestivum L., Santolina sp., Rosa sp., and Kochia scoparia (L.) Schrad. in the absence of orthokinetic effects. Experiments that excluded light and maintained relative humidity at 100% showed N. phyllobia to ascend the stems of 35 plant species to a height of > 9 cm within 12 h. Differences in stem ascension were not attributable to stem surface characteristics. PMID- 19305533 TI - Effect of Oryzalin and 1,1-Dimethylpiperidinium Chloride on Cotton and Tomato Roots Infected with the Root-knot Nematode, Meloidogyne incognita. AB - Oryzalin (3,5-dinitro-N4,N4-dipropyl-sulfanilamide) and BAS 083 (l,l dimethylpiperdinium chloride) reduced root-knot infection in tomato roots when respectively applied as a soil drench at 20 ppm and 10,000 ppm. Oryzalin reduced knot counts with various intervals between treatment and inoculation. BAS 083 reduced knot counts only when applied before inoculation. Oryzalin was shown not to be a contact nematicide, and BAS 083 was only a weak one. Neither compound reduced penetration by infective larvae. Postinfection reduction in knot counts by Oryzalin and BAS 083 resulted, in part, from activation of natural defense mechanisms of the host. Giant-cell development in cotton roots inoculated with nematodes was inhibited by Oryzalin. Lateral root development was inhibited by BAS 083. PMID- 19305534 TI - Effects of management practices on nematode and fungus populations and cucumber yield. AB - Three crops of cucumber were grown in succession in beds by use of trickle irrigation, plastic film mulch, and soil chemical treatments over a 17-month period, including a fallow winter season. Total yield for the three crops was highest (1208 quintals/ha) in film-mulched plots treated with MBR-CP, and next highest in film-mulched plots treated with DD-MENCS (1094 quintals/ha); total yield was only 456 quintals/ha in film-mulched control (untreated) plots. Yield in untreated film-mulched plots was 256% of that in untreated unmulched plots (178 quintals/ha). Plant growth and yields were greatest when populations of nematodes and soil-borne fungi were suppressed to very low levels. The residual control by soil treatments lasted longest on Meloidogyne incognita and Fusarium solani. PMID- 19305535 TI - The role of cuticular strata nomenclature in the systematics of nemata. AB - A system of cuticular nomenclature based on the strata observed in Enoplia is proposed. Nematode cuticle is divided into four fundamental strata: epicuticle, exocuticle, mesocuticle, and endocuticle. Application of this system allows the correlation of complementary strata throughout Nemata. The major taxonomic categories within Nemata are differentiated on the basis of their cuticular strata as compared with the Enoplia model cuticle. PMID- 19305536 TI - Techniques for Detecting Dactylella oviparasitica and Evaluating its Significiance in Field Soils. PMID- 19305538 TI - A method of distinguishing between living and dead nematodes by enzymatically induced fluorescence. PMID- 19305537 TI - Major- and Trace-Element Analyses of a Mermithid Parasite and its Mosquito Host by Proton-induced X-ray Emission. PMID- 19305539 TI - pH as a Factor in Parasitism of Mosquito Larvae by the Mermithid Romanomermis culicivorax. PMID- 19305540 TI - Equipment and techniques for establishing field microplots for the study of soilborne pathogens. PMID- 19305541 TI - New Data for Leptonchus transvaalensis from Nigeria, and Key to Leptonchus. PMID- 19305542 TI - Immune Responses of Mosquitoes against Romanomermis culicivorax (Mermithida: Nematoda). PMID- 19305543 TI - The Histopathological Reactions of Vigna sinensis to Separate and Concomitant Parasitism by Meloidogyne javanica and Rotylenchulus reniformis. AB - Cellular alterations in cowpea roots and nodules induced by single and concomitant Meloidogyne javanica and Rotylenchulus reniformis were investigated. M. javanica induced giant cells inside the vascular bundles of roots and nodules, and syncytia in cortical tissue of the nodules. In contrast, R. reniformis stimulated hypertrophy of pericycle and endodermal cells of the roots and nodules. Syncytia induced in the roots involved a sheet of pericycle cells and an endodermal cell. Cortical ceils of nodules also responded to R. reniformis infection, initiating wall gaps that led to syncytial formation. Coincidence of giant cells or syncytla of both nematodes was observed either in one vascular bundle or in separate ones. The histopathology of roots and nitrogen nodules infected by the two species remained unique even when they were feeding in close proximity. R. reniformis induced characteristic syncytia and M. javanica induced giant cells. PMID- 19305544 TI - The Relationship Between Population Density of Heterodera schachtii, Soil Temperature, and Sugarbeet Yields. AB - In two glasshouse experiments, relations between sugarbeet root dry weight (y, expressed as a percentage of the maximum dry root weight), and preplanting populations of Heterodera schachtii (P(i)) were described by the equation y = 100(Z)(P(i)-T), in which Z = a constant slightly smaller than 1, and T = the tolerance limit (the value of P(i) below which damage was not measureable). T varied with temperature; it was 65 eggs/100 g soil at 23 and 27 C and 430 eggs/100 g soil at 19 C. At 15 and 31 C there was no loss of root dry weight up to the maximum preplanting populations tested. In a field experiment in the Imperial Valley the relation between root yield (y) and P(i) was y = 100 (0.99886)(P(i) ) - 100, and the tolerance limit was 100 eggs/100 g soil. PMID- 19305545 TI - Pathogenicity of Hirschmanniella oryzae, H. spinicaudata, and H. imamuri on Rice. AB - In greenhouse experiments Hirschmanniella oryzae, H. imamuri, and H. spinicaudata depressed and delayed the tillering and flowering of rice, and suppressed root and shoot growth and grain yield. PMID- 19305546 TI - Interaction of Population Levels of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita on Cotton. AB - In autoclaved greenhouse soil without Fusarium oxysporum f. sp. vasinfectum, Meloidogyne incognita did not cause leaf or vascular discoloration of 59-day-old cotton plants. Plants had root galls with as few as 50 Meloidogyne larvae per plant. Root galling was directly proportional to the initial nematode population level. Fusarium wilt symptoms occurred without nematodes with 77,000 fungus propagules or more per gram of soil. As few as 50 Meloidogyne larvae accompanying 650 fungus propagules caused Fusarium wilt. With few exceptions, leaf symptoms appeared sooner as numbers of either or both organisms increased. In soils infested with both organisms, the extent of fungal invasion and colonization was well correlated with the extent of nematode galling and other indications of the Fusarium wilt syndrome. PMID- 19305547 TI - Chemical Control of Nematodes and Soil-borne Plant-Pathogenic Fungi on Cabbage Tranplants. AB - Six general-purpose fumigants and one fungicide were applied by different methods and evaluated for control of nematode-fungus complexes on cabbage grown for transplant production. All chemicals reduced populations of nematodes and soil borne fungi but varied greatly in effectiveness. Methyl bromide + chloropicrin (98% methyl bromide + 2% chloropicrin) (MBR-CP gas), DD + methyl isothiocyanate (DD-MENCS), methyl bromide + chloropicrin (67% methyl bromide + 31.75% cbloropicrin) (MBR-CP gel), and chloropicrin were more effective than sodium methyl dithiocarbamate (metham), pentachloronitrobenzene (PCNB), and potassium N hydroxy-methyl-N-methyldithiocarbamate (Bunema) against Meloidogyne incognita. Populations of Pythium spp. and Fusarium spp. were reduced markedly by all treatments except PCNB. Plant growth, uniformity, and yield were greater when nematodes and fungi were controlled. PMID- 19305548 TI - Granular Nematicides as Adjuncts to Fumigants for Control of Cotton Root-knot Nematodes. AB - Growth and yield of cotton were best with combinations of fumigants and organophosphate and carbamate nematicides. Organophosphates or carbamates used alone did not give season-long control of root-knot nematodes. Long-term control was poor because the temporary sublethal effects of these materials diminished soon enough lhat the nematodes could reproduce. The nematodes survived the treatments and a year of nonhost culture, and damaged a susceptible host crop 2 years after treatment. No such damage occurred in plots treated with fumigant, fumigant plus organophosphate, or fumigant plus carbamate. Treatment of seed and treatment of cotton, either in furrow at planting or sidedressing at midseason, with organophosphate and carbamate nematicides resulted in little or no yield increase, because nematode control was only minimal and temporary; or in a yield decrease, because the toxicity of the materials was manifested when nematode populations were low. PMID- 19305549 TI - Effects of nematicide placement on nematode populations and soybean yields. AB - Four methods of placement of DBCP (l,2-dibromo-3-chloropropane) and a single method of application of ethoprop (0-ethyl S,S-dipropyl phosphorodithioate) wexe compared in each of two areas for control of nematodes on soybeans. One area was a Marlboro sand infested with Hoplolaimus columbus. The other area was a Fuquay loamy sand infested with Meloidogne incognita. Soybean yields were increased and numbers of H. columbus in the row 0-20 cm deep were decreased similarly by all methods of DBCP application in Marlboro soil. All DBCP treatments increased the average soybean yields and decreased numbers of M. incognita larvae in the row 0 20 cm deep in the Fuquay soil. Average root-knot indices were reduced by all DBCP treatments except with placement 40 cm deep beneath the row. Similarly, placement of all or part of the DBCP 20 cm deep and 13 cm to either side of the row resulted in greater average yields than placement of the DBCP 40 cm deep. Apparently, control of M. incognita is more critical 0-20 cm deep than 20-40 cm deep for increasing soybean yields. DBCP did not control H. columbus as effectively as it did M. incognita. Control of H. columbus and M. incognita was not obtained at 0-20-cm and 20-40-cm depths 30 cm and 45 cm from the row regardless of the method used to apply DBCP. H. columbus and M. incognita were controlled more effectively and soybean yields were higher with DBCP at 13.6 kg a.i./ha than with ethoprop at 4.5 kg a.i./ha. PMID- 19305550 TI - Effects of Meloidogyne incognita on Nitrogen Fixation in Soybean. AB - The effects of a North Carolina population of Meloidogyne incognita on N fixation on root-knot-susceptible 'Lee 68' and moderately resistant 'Forrest' soybean were evaluated 50, 75, I00, and 135 days after inoculation with nematodes. Nematodes stimulated N fixation in Lee 68 by 50 days and in Forrest by 75 days. At all other intervals, N fixation was either depressed or unaffected by nematodes. Additional observations indicate that the susceptibility of Lee 68 is associated with greater rates of penetration by larvae and more favorable responses of host tissues to nematodes than occur in Forrest. With time, however, the histological reactions of both hosts became less favorable for nematode development. Resistant or hypersensitive responses became common in Forrest by 75 days but not in Lee 68 until 90 days after inoculation. This population of M. incognita may stimulate N fixation at a specific time interval and depress it at others; therefore, disease of susceptible soybeans caused by this nematode is probably not primarily due to a net loss of fixed nitrogen but to pathogenicity similar to that which occurs on nonlegume hosts. PMID- 19305551 TI - Growth Reduction of Apple Seedlings by Pratylenchus penetrans as Influenced by Seedling Age at Inoculation. AB - Apple seedlings of different ages (1, 3, and 5 weeks) were inoculated with 6,900 Pratylenchus penetrans per seedling in 10-cm-diam pots in a growth chamber. Rate of growth suppression and total growth suppression of seedlings by P. penetrans were inversely proportional to seedling age at time of nematode inoculation. Younger seedlings were found to contain a higher number of nematodes per gram root. PMID- 19305552 TI - Effect of Soil Water Potential on Growth of Apple Trees Infected with Pratylenchus penetrans. AB - Malling-Merton 106 apple rootstocks inoculated with Pratylenchus penetrans, or uninoculated, were grown in a growth chamber in pots of loamy sand maintained at two moisture levels, 0 to -0.4 bar or 0 to -10 bars. Either inoculation or low soil moisture suppressed shoot growth and increased root necrosis. However, the nematode-soil moisture interaction was not significant. PMID- 19305553 TI - Quantitative Aspects of the Development of Meloidogyne arenaria Larvae in Grapevine Varieties and Rootstocks. AB - The development and productivity of parasitic stages of Meloidogyne arenaria were quantitatively defined in 14 varieties or rootstocks of grapevine. Mean development to maturity was related linearly to the number of degree-hours above 10 C temperature experienced from the time of penetration in all cultivars in which nematode adulthood was achieved. Averaged across varieties, 13,142 heat units were required for development of the mean individual to maturity. The standard deviation of the developing individuals about the mean, expressed as a proportion with 1 representing adulthood, did not differ with time or among varieties after 7,000 degree-hours had elapsed. Earliest egg production was observed after 7,662 degree-hours, averaged across varieties, considerably before mean development to maturity. Varieties were also ranked relative to the number of larvae establishing infection sites, and the rate of egg production per adult female. Varieties could he grouped according to their levels of horizontal resistance. PMID- 19305554 TI - Descriptions of Meloidogyne camelliae n.sp. and M. querciana n.sp (Nematoda:Meloidogynidae), with SEM and Host-Range Observations. AB - Meloidogyne camelliae n.sp. on camellia (Camellia japonica) from Japan and M. querciana n.sp. on pin oak (Quercus palustris) from Virginia, USA, are described and illustrated. M. camelliae n.sp. is distingnishable from other species of the genus especially by its striking perineal pattern having heavy ropelike striae forming a squarish to rectangular outline with shoulders or projections, appearing sometimes ahnost starlike. M. querciana differs from other species by its characteristic perineal pattern round to oval in outline, sometimes with a low arch, and sunken vulva surrounded by a prominent obovate area devoid of striae. M. querciana shows some relationship to M. ovalis, but differs further fxom the latter by longer larvae, absence of annules on head of larvae, and rarity of males. Examination of specimens of M. camelliae n.sp. and M. querciana n.sp. with the scanning electron microscope confirmed observations made by optical microscopy and revealed diagnostic and other structures in greater detail. In greenhouse host tests, M. camelliae infected camellia heavily, showed moderate infection on oxalis, only a trace infection on tomato, and no infection on five other plants tested; and M. querciana attacked pin oak, red oak, and American chestnut heavily, but did not infect nine other test plants. In another test, pin oak seedlings did not become infected when heavily inoculated with and grown in the presence of two populations of M. incognita incognita and one of M. incognita acrita. The common names "camellia root-knot nematode" and "oak root knot nematode" are respectively proposed for M. camelliae and M. querciana. PMID- 19305555 TI - Anhydrobiotic coiling of nematodes in soil. AB - Nematodes of three genera (Acrobeloides sp., Aphelenchus avenae, and Scutellonema brachyurum) were induced to coil and enter anhydrobiosis in drying soil of two types: sandy loam and loamy sand. Coiling was studied in relationship to soil moisture characteristics. Coiling and the physiological state of anhydrobiosis occurred before the water in sandy soils reached a water potential of -15 bars. Coiling was maximum at 3-6 bars, depending on the soil type and nematode species. It appeared that induction of coiling and anhydrohiosis were determined by the physical forces exerted by the water film surrounding the nematode, which, for these three species, was 6-9 monomolecular layers of water, rather than the % moisture and relative humidity of the soil per se. PMID- 19305556 TI - Modification of the Mini-sieve and Prepolymerized Plate Techniques for Use in Electron Microscopy. PMID- 19305557 TI - Influence of Plant Age, Light Intensity, Nematode Inoculum Level, and Their Interactions on Tomato Growth and Reproduction of Meloidogyne hapla. PMID- 19305558 TI - Technique for Screening Cowpea Germplasm for Resistance to Root-knot Nematodes. PMID- 19305559 TI - A rapid method for sampling surface soil. PMID- 19305561 TI - Automated dehydration of small specimens. PMID- 19305560 TI - Chemical Control of Heterodera schachtii on Sugarbeet in California. PMID- 19305562 TI - Nonsolanaceous hosts of globodera in the andes. PMID- 19305563 TI - Historical development of nematology in Russia. AB - The development of Russian hematology is considered from the late nineteenth century to 1970. The dominant influences of I. N. Filipjev and A. A. Paramonov are discussed in the context of the persons whom they influenced and their conceptual approach to the problems posed hy nematodes. The advantages and disadvantages of the framework of Russian scientific administration are compared to those in the West. PMID- 19305564 TI - World dissemination of the cereal-cyst nematode (Heterodera avenae) and its potential as a pathogen of wheat. AB - World distribution of the cereal-cyst nematode is herein reviewed. It is suggested that Heterodera avenae originated in Europe and has been widely disseminated, largely by the activities of Man but also by wind movement of cysts. So far, it may not have spread to some major wheat-growing regions of the New World, but a non-friable soil structure limits population level and disease. Yield loss could result from the introduction of new cultivars to developing countries where H . avenae has not been detected or where existing cultivars possess tolerance. PMID- 19305565 TI - Worldwide dissemination of Radopholus similis and its importance in crop production. AB - The burrowing nematode, Radopholus similis, attacks agronomic and horticultural crops and many weeds, and is reported to reproduce on more than 250 plant species. Two races of R. similis are recognized. Although one race attacks citrus and the other race does not, they are morphologically similar. At present, the citrus race is found attacking citrus only in Florida, U.S.A., but it is known to infect more than 250 species and varieties of noncitrus plants. Although it has many hosts, R. similis is probably most widely distributed on banana and is found worldwide. Although best known as a pest of Piper nigrum, Musa spp., and Citrus spp., it also attacks many crops that are important in world commerce and in subsistence-type agriculture, a factor which makes it a significant agricultural pest. Worldwide dissemination occurs primarily when parasitized plants are moved into areas where the pest could adapt. Yield losses of 12.5 tons/ha in bananas have been reported from R. similis infection. Infections suppress orange and grapefruit yields as much as 70-80%. Because of the severity of R. similis damage (particularly to banana and citrus), extensive control programs have been developed. Prevention, cultural practices, resistant varieties, and chemical pesticides interact to reduce losses. PMID- 19305566 TI - Worldwide dissemination and importance of the root-knot nematodes, Meloidogyne spp. AB - Root-knot nematodes are widely distributed throughout the world. Their distribution and economic importance are purported to be related to biological and environmental factors favorable to Meloidogyne spp. A scheme for comparing Meloidogyne spp. with two other genera of diverse biological characteristics is presented to support this hypothesis. It is further suggested that a probability index can be developed to predict the likelihood of a given nematode species being transported, established, and becoming economically important in regions of the world where i t does not already occur. PMID- 19305567 TI - Worldwide distribution of potato-cyst nematodes and their importance in crop production. AB - The potato-cyst nematodes Heterodera rostochiensis and H. pallida are important pathogens of potatoes, a basic food crop. When soil populations of either species are high, potato yields are often less than the seed planted. Apparently, these nematodes originated in the Andean mountains of South America, the home of the potato. One or both species have spread from this region to approximately 47 countries. The two species may occur together or separately in potato-growing areas. Although these nematodes can be spread in numerous ways, contaminated soil associated with seed potatoes, farm machinery, and reusable containers is among the most important. An integrated control program used in the U.S.A. is described. PMID- 19305568 TI - Worldwide distribution of soybean-cyst nematode and its economic importance. AB - Soybean-cyst nematode (Heterodera glycines) was first reported from Japan in 1915. It has since been reported from Korea (1936), Manchuria (1938), the United States (USA) (1954), and Egypt (1968). It is of major concern to soybean producers only in Japan and the USA. Soybean was a major crop in the Orient by 1915 but it was grown very little elsewhere. Since that time its cultivation has spread, and in 1974 there were 37.6 million ha with a production of 51.7 billion kg. As soybean cultivation has spread, the soybean-cyst nematode has been more widely found. Soybean is one of the major food sources for feeding the increasing world population. Soybean-cyst nematodes have been spreading rapidly in recent years and are a major threat to this very important crop. PMID- 19305569 TI - Infectivity of Bacillus penetrans in plant-parasitic Nematodes. AB - Larvae of Meloidogyne spp. were readily infected with the endoparasite Bacillus penetrans by exposure to an aqueous suspension of spores from infected root-knot nematode females, or by passage of larvae through a shallow layer (50 cm(3)) of spore-infested soil. Infection severely reduced motility of second-stage larvae through soil. Bacillus penetrans exhibited a distinct host specificity in that only 5 of 16 nematode species tested became infected with the population used. Meloidogyne javanica, M. arenaria, and M. incognita became more heavily infected than M. hapla or Pratylenchus scribneri under similar conditions, but all of these species were also good hosts. Ultrastructural changes in the endospore within sporangia adhering to the cuticle are similar to those reported for other germinating Bacillus endospores. PMID- 19305570 TI - Control of citrus nematode Tylenchulus semipenetrans on fine-textured soil with DBCP and oxamyl. AB - Three grapefruit orchards on sour orange rootstock were treated by metering DBCP (l,2-dibromo-3-chloropropane) at 56 kg(a.i.)/ha into 15 cm of water in a flood irrigation system. In orchards with 43-49% clay in the surface foot, DBCP reduced numbers of Tylenchulus semipenetrans below control levels for 1.5-2.0 years. In the 3 orchards, DBCP treatment resulted in increases in yield or fruit size in the 2 seasons following treatment. No increases in yield, fruit size, or fruit numbers were observed the third season after treatment, but retreatment of a portion of one orchard after 2 years resulted in large increases in yield and fruit numbers the following harvest. Application of oxamyl twice annually as a foliar spray at 2.8 kg(a.i.)/ha reduced nematode populations to about 50% of control levels and resuhed in a large increase in yield in 1 of the 2 seasons tested. DBCP treatment of fine-textured soils controlled citrus nematode and increased yields, but its effect was not as long lasting as on coarser soil in other citrus areas. Foliar applications of oxamyl reduced nematode populations but were not as consistently effective as DBCP treatments. PMID- 19305571 TI - Penetration and development of Meloidogyne hapla in resistant and susceptible alfalfa under differing temperatures. AB - Studies were conducted to examine under differing temperatures (12, 16, 20, 24, 28, and 32 C) the penetration anti development of Meloidogyne hapla in resistant lines '298' and 'Nev. Syn XX', and susceptible 'Lahontan' and 'Ranger' hardy-type alfalfas. The results indicated that resistance to M. hapla was similar to that previously described for M. incognita in nonhardy alfalfa. Although initial penetration in resistant seedlings was similar to that of susceptible seedlings, nematode larvae failed to establish and develop in root tissues and nematode numbers subsequently declined. In susceptible seedlings, nematode development proceeded rapidly, and egg production began after 5 weeks. Temperature had little influence on the nematode development except to slow the response at the lower temperatures. Other studies were conducted to verify a previously reported immune (no penetration) reaction to M. hapla by the 'Vernal' selection 'M-4'. When compared to the resistant (penetration without nematode development) Vernal selection 'M-9' under differing temperatures (20, 24, 28, and 32 C), each selection was equally penetrated by M. hapla but at a lower level than in susceptible Ranger cuttings. Generally, no root galling was observed in either M 4 or M-9; however, very slight galling was found 35 days after inoculation on about 50% of these cuttings when grown at 32 C. PMID- 19305572 TI - Ultrastructure of the anterior body region of marine nematode Deontostoma californicum. AB - The ultrastructure of the anterior body region of the free-living marine nematode Deontostoma calilornicum was studied by electron microscopy. The body wall consists of a nine-layered cuticle, a cellular hypodermis containing eight nerve bundles, and a well-developed coelomyarian somatic musculature. Nerves in the dorsal, lateral, ventral, and submedian hypodermal chords anterior to the nerve ring were observed with regularity. Structure of subventral somatic setae suggests a mechanoreceptive function. The esophagus is cellular and consists of three marginal cells alternating with an equal number of radial muscle cells, three esophageal glands, and three enteric nerves. The membranes of adjacent esophageal cells are sinuous. Apices of the triradiate lumen are connected with the outer wall of each marginal cell by bands of electron-dense nonmyofibrils, whereas two types of myofilaments run radially between the apophyses of the lumen and the outer walls of radial cells. Each myofibril, which forms hemidesmosomes at both ends, is interpreted to be the morphological equivalent of one sarcomere. Synaptic junctions between the processes of muscles, gland cells, and axons of the enteric nerves are described in detail. PMID- 19305573 TI - Effects of subsoiling and nematicides on Hoplolaimus columbus populations and cotton yield. AB - Subsoiling to a depth of 35 cm under the planting row for 3 consecutive years increased annual yields of seed cotton by 50 to 200%. Annual subsoiling was essential for maximum yields. The application of a nematicide, 1,2-dibromo-3 chloropropane (DBCP) or aldicarb, reduced the population of Hoplolaimus columbus but did not increase seed-cotton yields over subsoiling alone. Subsoiling reduced H. columbus in the top 20 cm of soil since the treatment favored deeper penetration by much of the root syslem and, consequently, less root colonization of the upper soil zone. PMID- 19305574 TI - Observations on the host-parasite relationship of Pratylenchus vulnus on grapevine, Vitis vinifera. PMID- 19305575 TI - Meloidogyne incognita: a metabolic sink. PMID- 19305576 TI - Heterodera thornei n. sp. (Nematoda:Heteroderidae) and a review of related species. AB - In a taxonomic review of the species of the Heterodera "cacti group," H. thornei n. sp. is described and illustrated from Miner's lettuce in California. This new species is distinguishable especially by havpunctate egg shells and a larval stylet (27 mum) and tail (56 mum) longer than other related species. Heterodera weissi is redescribed, type specimens are designated, and new data added on the basis of original and topotype specimens. New morphometric data, along with illustrations, are given for H. cacti, H. amaranthi, and H. estonica, and certain details in the original descriptions are clarified on the latter two species. Diagnoses and a key for identification of these species are given. A common name for each species is proposed when one is lacking. Previously unrecognized tooth like structures occurring in the vulval cone top of these species and in H. schachtii are characterized and named "vulval denticles." Because of its markedly different morphology, especially in cysts, H. betulae is removed from the H. cacti group and placed alone, since it is incompatible with any of the present Heterodera "groups." Know distribution of these species is given, with the result that many new areas and countries of occurrence for some of the species are listed. PMID- 19305577 TI - The nature and role of metabolic leakage from root-knot nematode galls and infection by Rhizoctonia solani. AB - A severe root rot of tomato caused by Meloidogyne incognita and Rhizoctonia solani was associated with nutrient mobilization into gall tissue and root exudation. Root decay did not develop when root exudates were continuously removed by leaching. When leachates were collected from M. incognita-infected and control roots and applied to roots of tomatoes inoculated with R. solani alone, roots receiving leachates from M. incognita-infected roots developed a severe rot while roots receiving leachates from control roots were free of decay. During the fourth and fifth weeks following nematode infection, an increased mobilization of (1)C labelled compounds to nematode-infected roots occurred. Higher counts of water soluble nonvolatile (1)C labelled exudates leaked from nematode-infected roots than from control roots at each weekly sampling period. Higher concentrations of Ca, Mg, Na, K, Fe, and Cu were found in exudates from nematode infected roots than from the control roots. During the first 14 days following nematode infection, carbohydrates were the major organic constituents in exudates leaking from nematode-infected roots. Fourteen days after nematode infection, nitrogenous compounds become the major organic constituents leaking from roots. Shifts in C/N ratio of root exudates from nematode-infected roots were associated with parasitic development of R. solani in tomato roots. PMID- 19305578 TI - The virus-vector and damage potential, morphometrics and distribution of Paralongidorus maximus. AB - Paralongidorus maximus was tested as a vector for 20 viruses. Nine were recovered directly from P. maximus, an indication that they had ingested the virus, but there was no correlation between the ingestion and transmission of these viruses. Raspberry ringspot (English and Scottish strains), arabis mosaic, and strawberry latent ringspot viruses were recovered from the roots of bait plants grown in pots together with virus infected plants and P. rnaximus. Under the experimental conditions, these four viruses may have been transmitted by P. rnaxirnus. Sixteen of these viruses were not transmitted by P. maximus. None were transmitted when P. maximus was extracted between the infector and bait plants. The authors conclude that P. maximus is not likely to be a vector, under field conditions, of any of the presently known nematode-transmitted viruses. A new site is reported for P. maximus in a forest nursery in Angus, Scotland where it was causing considerable damage to 3- and 4-year-old Scots pine (Pinus sylvestris). Morphometrically, this population (which was used in the virus tests) is very similar to populations from Germany and other parts of Britain. Geographical distribution of this species is restricted to western Europe, where i t is widely distributed, and to only three sites in the British Isles. PMID- 19305579 TI - Host-parasite interactions of Pratylenchus scribneri on selected crop plants. AB - Greenhouse tests were conducted to determine the effects of soil temperature and texture on development of Pratylenchus scribneri and the pathogenicity and reproductive rates of this nematode on selected crop plants. In a sandy loam soil, greatest numbers of P. scribneri were found at 30 and 35 C on sudangrass and sugarbeet, respectively. In a silty clay loam, the nematode reproduced best at 35 C on sugarbeet. Higher populations of P. scribneri were found in the sandy loam than silty clay loam soil at corresponding temperatures. In a pathogenicity test, top and root growth of sudangrass and barley were suppressed by the nematode, whereas no significant growth inhibition was found on wheat and alfalfa. Tests with other vegetable and field crops indicated wide variance in nematode reproduction. PMID- 19305580 TI - Effects of management practices on nematode and fungi populations and okra yield. AB - Okra was grown in field plots of Tifton loamy sand naturally infested with the nematodes Meloidogyne incognita and Criconemoides ornalus and the pathogenic fungi Fusarium oxysporum, F. solani, F. roseum, and Pythium spp. Plots were treated with various soil pesticides and left exposed or covered with biodegradable paper film mulch under trickle irrigation. Soil was assayed for nematodes and fungi, and plant roots were examined for root-rot and insect damage. Fewer nematodes and fungi generally were recovered from soil treated with DD-MENCS (with and without film mulch) or methyl bromide-chloropicrin (2:1) (MBC) and film mulch than from nontreated soil. Funfigation with DD-MENCS or MBC suppressed populations of M. incognita, C. ornatus, F. oxysporum, F. solani, F. roseum, and Pythium spp. Ethoprop (alone or combined with other pesticides), sodium azide, and chloroneb were less effective than DD-MENCS and MBC. Plant growth anti yield were greatest when nematodes and pathogenic fungi were controlled. Yield was increased 3-fold by DD-MENCS + film mulch or MBC + film mulch in comparison with the average yield of okra produced in Georgia. The root knot nematode-Fusarium wilt complex was most severe in nonfuntigated soil. PMID- 19305581 TI - Reproductive biology and behavior of Rhabditis pellio, (Schneider) (Rhabditida: Rhabditidae). AB - Laboratory studies were conducted on the mating behavior of Rhabditis pellio males and females, which were maintained on a culture of Flavobacterium sp. bacteria isolated from earthworms. The mean time that elapsed between first contact of the sexes and their ultimate separation was 23.2 min. However, only 5.0 min were required for copulation (the interval during which male spicules were inserted into the female vagina). Three-day-old females that were permitted to mate once on their first day of adult life produced only one-third as many larvae as did females that were permitted unlimited mating. However, the longevity of females was found to decrease with an increase in the number of matings. Both males and females that were permitted to mate daily produced the greatest number of offspring when they were 4 days old. When the initiation of mating was delayed beyond their third day of life, the number of larvae produced by females decreased. In approximately one-half of the copulations, males failed to inseminate their female partners. PMID- 19305582 TI - Effects of selected carbamate and organophosphate nematicides on hatching and emergence of Heterodera schachtii. AB - Ethoprop, oxamyl, PP 156, fenamiphos, carbofuran , AC 64,475, Bunema M, CG 12223, aldicarb, aldicarbsulfoxide, and aldicarbsulfone were tested for their effects on hatching and emergence of larvae from cysts of Heterodera schachtii. The oxime carbamates and carbofuran inhibited hatching , but this response was reversed by removing the chemical treatment . Inhibition of hatching by Bunema M and all organophosphates tested was irreversible . PMID- 19305583 TI - Effects of Pratylenchus vulnus on the growth of sour orange. AB - Pratylenchus vulnus suppressed the growth of sour orange seedlings in greenhouse experiments. Growth retardation (in height, in trunk diameter, and in dry top and root weights) was observed in inoculated plants growing in two soil types. Population density, 13 months after inoculation, averaged more than 1,000 specimens/gm of fresh root. Anatomical studies showed that P. vulnus prefers to attack the cortex and causes cavities among the cortical cells. PMID- 19305584 TI - Nematicidal effects of oxamyl applied to leaves of banana seedlings. AB - Foliar applications of oxamyl prevented nematodes from invading roots of diploid bananas. One spray with 1,250 microg/ml was more effective than 1, 2, or 3 sprays with 625 microg/ml applied at 5-day intervals. After 3 sprays with 1,250 microg/ml, invasion may be prevented for up to 4 weeks and possibly longer. Washing roots after oxamyl treatments prevented nematicidal control. When applied to nematode-infected plants, three sprays of oxamyl decreased nematode populations in the roots. PMID- 19305585 TI - Comparison of five populations of Tylenchulus semipenetrans to Citrus, Poncirus, and their hybrids. AB - The infectivity of five populations of Tylenchulus semipenetrans were compared and differentiated on 10 hosts (5 Citrus spp., 1 Poncirus trifoliata, and 4 hybrids of Citrus spp. X P. trifoliata). Differences in levels of infection and development (P = 0.01) occurred between Citrus spp. and P. trifoliata cv. 'Pomeroy' and their three hybrids, C. paradisi X P. trifoliata cv. 'Swingle' citruntelo and C. sinensis, cv. 'Ruby' orange X P. trifoliata cv. 'Webber Fawcett 14-7', and '15-7'. Poncirus trifoliata cv. Pomeroy was susceptible to a California biotype 3 and highly resistant to the other citrus nematode populations. Low infection levels with California biotype 1, Arizona, and Florida populations on Swingle citrumelo, and the two Ruby orange hybrids indicated inherited resistance. Reproduction of the nematode population from Texas was greatest on the three hybrids, Swingle citrumelo, Ruby orange 14-7, and 15-7, from the California 1, Arizona, and Florida populations, but its comparable densities on P. trifoliata and Citrus spp. were not sufficiently different from these populations to consider it a separate biotype. California biotype 3 was sufficiently different from all other populations to be considered a different biotype, and it was named the "Poncirus biotype." PMID- 19305586 TI - The effects of temperature on the infectivity of Romanomermis culicivorax. AB - The survival time (ST) of the preparasitic larvae of Rornanomermis culicivorax was determined by measuring motility at 1, 6, 12, 18, 21, 27, 30, and 37 C; the ST at each of these temperatures was 2.3, 2.2, 2.0, 2.0, 1.7, 1.6, 0.9, and 0.7 days, respectively. About one-third of the preparasites infected first-instar larvae of Culex pipiens within 24 h at 27 C. The preparasites were infective at 12 to 33 C with the optimum infectivity at 21-33 C. Lower temperatures decreased the percent infectivity but increased the time that the nematodes remained infective. The time required for host infection increased as the preparasitic larvae aged at 15, 21, and 27 C. PMID- 19305587 TI - Temperature effects on the parasitic phase of Romanomermis culicivorax in Culex pipiens. AB - The developmental period for the parasitic stage of Romanomermis culicivorax in Culex pipiens was determined at constant and fluctuating temperatures. The median developmental times at 15, 18, 20, 27, and 32 C were 27.6, 17.2, 11.5, 7.1, and 5.8 days, respectively. The optimum temperature range for development of the parasitic stage in C. pipiens was 20 to 32 C. The threshold for development was calculated as 10.4 C, and the heat units required for development were 122.2 +/- 3.6 day-degrees. Development at fluctuating temperatures conformed to that predicted by the constant temperature data. PMID- 19305588 TI - A comparison of techniques for extraction and study of anhydrobiotic nematodes from dry soils. AB - Anhydrobiotic nematodes were fixed and extracted from dry Mojave desert soils with hot and cold fixatives (5% formalin and 4% gluteraldehyde). Morphologically, extracted nematodes were tightly coiled and shrunken in size. Various concentrations of KCl, ethylene glycol, and sucrose solutions were compared for their effectiveness in extracting viable nematodes still in the anhydrobiotic state. Approximately 80-95% of the anhydrobiotic nematodes extracted with 1.25 M and 1.5 M sucrose were tightly coiled and shrunken in a manner similar to those fixed and extracted in formalin. Anhydrobiotic nematodes can be maintained in this form for up to 24 days in 1.25 M sucrose solution and still be revived. All molarities of ethylene glycol and KC1 tested were ineffective in recovering and maintaining nematodes in the natural anhydrobiotic state. Straight, air-dried, and active nematodes served as controls and did not coil when placed in hot or cold fixatives or in any concentration of KCI, ethylene glycol, or sucrose. Anhydrobiosis, as represented by the coiled form of nematodes from desert soils, was not confined to any particular life stage or trophic group. PMID- 19305589 TI - Suppression of Pratylenchus penetrans and Tylenchorhynchus dubius by Trichoderma viride. PMID- 19305590 TI - A possible new approach to the chemical control of nematodes. PMID- 19305591 TI - Pectolytic enzymes in three populations of Ditylenchus dipsaci. AB - Extracts of nematodes of the Raleigh, North Carolina (RNC), Waynesville, N. C. (WNC), and onion populations of Ditylenchus dipsaci were examined for pectolytic activity. RNC nematodes contained a NaCl-stimulated endo-polymethylgalacturonase with optimal pH for activity of 6.0, whereas nematodes of the WNC and onion populations possessed a NaCl-stimulated endo-polygalacturonase with pH optimum of 4.0. Nematodes of each population also contained a CaCl-activated endo-pectin methyl-trans-eliminase with optimal pH of 9.0. Nematode extracts containing 0.5 M NaCl macerated potato discs. RNC and onion nematodes induced gall formation in Wando pea seedlings, but WNC nematodes induced a resistant, hypersensitive response. Thus pectolytic activity was not correlated with pathogenicity of D. dipsaci on Wando pea. PMID- 19305592 TI - Effects of hydroclytic enzymes on plant-parasitic nematodes. AB - Proteases, lipase, and chitinase killed Tylenchorhynchus dubius in vitro and in soil. Tylenchorhynchus dubius was more susceptible to the enzymes than Pratylenchus penetrans. Papain was the most effective protease, and other enzymes were less effective. Heating enzymes to 80 C for 10 min greatly reduced nematicidal effectiveness. Scanning electron micrographs showed that papain and chitinase produced structural changes in the cuticle of T. dubius. Lipase removed a thin outer layer. Papain removed material filling the striata, or furrow, between the horizontal bands. When added to soil, chitinase, lipase, collagenase, and proteases (papain and bromelain) decreased motility of T. dubius populations up to 75%. Bromelain was the most active in soil against T. dubius, and collagenase was the most active in soil against P. penetrans. PMID- 19305593 TI - Species differentiation in Caenorhabditis briggsae and Caenorhabditis elegans. AB - Identification of five laboratory strains (1-5) of putative Caenorhabditis briggsae was undertaken. Examination of the male bursal ray arrangement, mating tests with males of Caenorhabditis elegans, malate dehydrogenase zymograms, and SDS polyacrylamide electrophoresis demonstrated that strain 4 was C. briggsae and the others were C. elegans. PMID- 19305594 TI - Nematicidal activity of the fungicide ethazole. AB - The nematicidal activity of the fungicide ethazole was studied under greenhouse conditions in which a nematode-infested, Norfolk sandy loam was planted to cotton (var. = 'Rowden'). At planting time, the fungicide was incorporated in the soil at rates of 0-100 mg/kg soil. When assayed 4 weeks after planting, plant parasitic nematodes in soil and roots occurred in decreasing numbers with increasing concentrations of the fungicide; the degree of control obtained was from 60-100% with the four highest concentrations. Results indicate that ethazole possesses nematicidal activity at rates recommended for field application. PMID- 19305595 TI - The sex ratio of Heterodera glycines at low population densities. AB - The sex ratio of the Arkansas 1 isolate of Heterodera glycines was determined in experiments in which 'Lee' soybean was inoculated with either one or two larvae. A 3:1 male to female sex ratio was established for this isolate under the test conditions used. No influence of one nematode on the penetration and development to adult of another nematode in the same root was detected in double larval inoculations. PMID- 19305596 TI - Effects of soil moisture on control of Heterodera schachtii with Aldicarb. AB - Soil moisture and the nematode population density in aldicarb-treated soil influenced control of the sugarbeet nematode, Heterodera schachtii. Greater numbers of nematode larvae infected 14-day-old sugarbeet seedlings growing in aldicarb-treated soil at 20-30% than at 80-100% field capacity (F. C.), and plant growth was inversely related to nematode infection and the nematode population density. Compared with that of control plants, plant growth increase also was greater at 80-100% F. C. when the nematode population was above 1.8 larvae/gm soil. A nematode population of 1.8 larvae/gm soil did not significantly affect sugarbeet yields. Aldicarb gave less control when soil moisture levels dropped to 20 and 50% F. C. at nematode populations of 3.5 and 6.2 larvae/gm soil. More effective control was obtained wth soil moisture levels at or above 80% F. C. This difference was attributed to continued activity of the toxicant in the rhizosphere at the high moisture level. PMID- 19305597 TI - Effects of Macroposthonia xenoplax on the growth of Concord grape. AB - Concord grape (Vitis labrusca) plants were inoculated with Macroposthonia xenoplax at levels of 100, 1,000, and 10,000 nematodes. After 4 months, plants inoculated with 10,000 M. xenoplax were stunted, and root systems were darker and had fewer feeder roots than those in other treatments. The lower nematode inoculation levels suppressed top growth but did not affect root growth. M. xenoplax reproduced well on Concord grapes. PMID- 19305598 TI - Effects of temperature and photoperiod on the infection of two mosquito species by the mermithid Romanomermis culicivorax. AB - Successful invasion by the mermithid Romanornermis eulicivorax declined linearly from 93.6 to 1.5% in Culex tarsalis and from 73.1 to 1.6% i n Aeries dorsalis larvae exposed in the laboratory at 18, 16, 14, 12, and l 0 C. Larvae of C. tarsalis were more susceptible than those of A. dorsalis at 18 and 16 C, but this relationship was reversed at 12 C. Larval mortality during the 48-h exposure period was due primarily to nematode infection . Photoperiod had no effect on infection. PMID- 19305599 TI - Influence of Nemaguard and Lovell rootstocks and Macroposthonia xenoplax on bacterial canker of peach. AB - 'Fay Elberta' peach trees grown on either 'Lovell' or 'Nemaguard' rootstocks in sandy soil in a lathhouse were highly susceptible to bacterial canker if inoculated with the nematode Macroposthonia xenoplax and the bacterium Pseudomonas syringae. If either one of these organisms were omitted, serious bacterial canker did not develop. Cankers appeared later and remained small when nematodes were omined. Very few cankers appeared on trees not inoculated with the bacterium. Peach trees on both rootstocks were good hosts for, and were stunted by, nematodes. Larger numbers of fruit were produced on trees free of bacterial canker or nematodes. Differences in magnitude of bacterial canker symptoms produced experimentally in different years are considered. PMID- 19305600 TI - Terpenoid aldehydes in cotton roots susceptible and resistant to the root-knot nematode, Meloidogyne incognita. AB - We investigated the role of terpenoid aldehydes in the resistance of cotton (Gossypium hirsutum) to the root-knot nematode (Meloidogyne incognita). Three-day old, root-knot-resistant ('Auburn 623') and -susceptible ('Deltapine 16') seedlings were inoculated with M. incognita. Comparable portions of inoculated and noninoculated roots were harvested 2, 4, 7, and 10 days later. Terpenoid aldehydes were extracted, separated by thin-layer chromatography, eluted as their phloroglucinol derivatives, and measured colorimetrically. In noninoculated seedlings of each age, the susceptible cultivar contained more total and more of each of five specific terpenoid aldehydes (hemigossypol, methoxyhemigossypol, gossypol, lnethoxygossypol, dimethoxygossypol) than did the resistant cultivar. In both cultivars, the concentration of terpenoid aldehydes increased as seedlings aged. After inoculation, the concentration of terpenoid aldehydes was usually highest in the noninoculated, followed by the infected susceptible, infected resistant, and noninfected resistant seedlings in that order. The changes in concentration that occurred in response to infection, particularly at 7 and 10 days after inoculation, did correlate with host resistance, i.e., there was a net loss of total and each specific terpenoid aldelhyde in tlae susceptible cultivar, and a net gain in the resistant. Our data do not exclude the possibility that localized synthesis of terpenoid aldehydes is involved in resistance to root-knot nematodes. PMID- 19305601 TI - Anguina plantaginis n. sp. parasitic on Plantago aristata with a description of its developmental stages. AB - Anguina plantaginis n. sp., parasitic on Plantago aristata, is described and illustrated. This new species is most closely related to A. klebahni, A. millefolii, A. mobilis, and A. moxae and is characterized as follows: moderate body size for the genus; absence of esophageal "storage organ"; postvulval uterine sac extending about 45% of vulva-anus distance; crustaformeria of young females longer than spermatotheca or uterus proper; spicules with 2 sclerotized thickenings; long, conical tail in both sexes, narrowing at about 1/6 of its length to peg-like tip; parasitic only on P. aristata. Two nematode generations that are morphologically similar but differ in body size develop in one plant gall. The postembryogenesis, studied with respect to morphological development of the larval stages, is similar to that of Ditylenchus. The sexes can be differentiated from the second molt on. The infective larva is the third stage, which is morphologically distinct from the regularly developing third-stage larva. PMID- 19305602 TI - New records of nematodes from Korea, including Paratylenchus pandus n.sp. (Paratylenchidae nematoda). AB - Five new records of Paratylenchus, including P. pandus n.sp., are reported from Korea. An amended key to the genus is included on the basis of these findings. Macroposthonia tulagonovi is also reported with additional descriptions and illustrations. PMID- 19305603 TI - Identification of field populations of Meloidogyne spp. by chromosome number. PMID- 19305604 TI - A technique for the collection of larvae of Meloidogyne spp. and a comparison of eggs and larvae as inocula. PMID- 19305605 TI - A comparison of two methods of synchronous inoculation of cotton seedlings with Meloidogyne incognita. PMID- 19305606 TI - Critical-oxygen tension of Caenorhabdiltis elegans. PMID- 19305608 TI - Phenol levels in leaves of tomato cultivars, infected with Paratylenchus penetrans. PMID- 19305607 TI - The face view of trichodoridae. PMID- 19305609 TI - Thermal tolerance of Romanomermis culicivorax, a nematode parasite of mosquitoes. PMID- 19305610 TI - Analysis of crop losses in tomato due to Pratylenchus penetrans. AB - The effects of Pratylenchus penetrans upon yields of 'Veebrite' tomato were studied at initial soil population densities (P(i)) of 360, 2,010, 4,580, and 14,360 nematodes/kg of soil in 20-cm (i.d.) clay-tile microplots. The lowest P(i) appeared to stimulate fruit production. Higher P(i)'s suppressed fruit production (total weight of marketable tomatoes and numbers of intermediate- and large-sized fruits), in comparison to control yields, the highest P(i) resulted in 38% fewer fruits which weighed 44% less. These losses were at least partly due to a delay in fruit ripening, caused by the nematodes, which did not become apparent until the fourth week. Nematode populations in the soil increased at all but the highest P(i); final populations were around 7,000/kg of soil. Nematode populations in roots ranged from 230-590/gm of root at the completion of the experiment. Nematode control by fumigation would definitely be warranted at soil population densities of 2,000/kg or higher; with 500-2,000/kg, the decision to fumigate would depend on soil type and economic and hiological factors. PMID- 19305611 TI - Effects of population densities of Meloidogyne hapla on growth and yield of tomato. AB - Growth and yield of 'Veebrite' tomato were studied in 20-cm (i.d.) clay-tile microplots containing initially 260, 1,840, 6,120, or 27,950 Meloidogyne hapla larvae/kg of soil. Low nematode numbers stimulated, and the highest nematode population suppressed, vegetative plant growth. More tomatoes, with a higher total weight, were harvested from plants infested with 260 and 1,840 nematode larvae at planting than from those with initial densities of 6,120 and 27,950 larvae. At the two highest densities, the cumulative fruit production (weight) was suppressed by 10% and 40%, respectively. The increase in growth and yield at the lower densities appeared to be due to an increase in the size of the root systent. However, at the higher densities, yield was no longer directly related to root weight. The reproduction factor of M. hapla was negatively correlated with initial density; for the lowest and highest initial densities, it was 96X and 7X at midseason, and 354X and 3X at harvest, respectively. The equilibrium density was 63,000 larvae/kg of soil; initial densities larger than 2,000 larvae/kg of soil may require control. PMID- 19305612 TI - Population growth patterns of four species of aphelenchoides on fungi. AB - Qualitative and quantitative differences in population growth patterns of Aphelenchoides rutgersi from Florida, A. sacchari from Jamaica, A. dactylocercus from Great Britain, and A. cibolensis from New Mexico were assessed on 28 species of fungi. The patterns of population growth of A. rutgersi and A. sacchari were statistically similar although not identical, and they differed considerably from those of A. dactylocercus and A. cibolensis. It is suggested that A. rutgersi and A. sacchari, from Florida and Jamaica respectively, may be more closely related to each other than to either A. dactylocercus or A. cibolensis. PMID- 19305613 TI - Nematicidal injection: targeted control of plant-parasitic nematodes of trees and vines. AB - Pressurized injection of nematicidal solutions was effective for control of nematodes within trees and vines. Significant (P = 0.01) control of Pratylenchus vulnus on grape was attained with four nonfumigants (carbofuran, oxamyl, phenamiphos, and sulfocarb) and one fumigant nematicide (DBCP). Pratylenchus penetrans was controlled (P = 0.05 and 0.1) in apples and walnuts with sulfocarh and oxamyl. This species also was controlled in apples with carbofuran and phenamiphos. The advantages of pressure injection over traditional methods of nematicide applications are discussed. PMID- 19305614 TI - Development of Heterodera glycines pathotypes as affected by soybean cultivars. AB - The reproductive abilities of four races of Heterodera glycines were compared on soybean cultivars by using single cyst or mass inoculations. Progeny transfers were used to determine changes in reproduction of H. glycines. Reproduction of all races (1, 2, 3, 4) was best on 'Lee' and poorest on P1 88788. The size of females produced and numbers of eggs/female of the different races varied with the cultivar. Races 1 and 3 appeared to contain low populations of Races 2 or 4. Races 2 and 4 were best selected by a series of transfers on 'Pickett' soybean. PMID- 19305615 TI - The use of glass microbeads in ecological experiments with bacteriophagic nematodes. AB - A system that uses microbeads for the culture of bacteriophgic nematodes is described. Glass-bead culture was found to simulate a soil microcosm more closely than did agar culture i n terms of CO production , number of nematodes produced, and nematode size. PMID- 19305616 TI - Evaluation of a seed-treatment method with acetone for delivering systemic nematicides with wheat and rye. AB - Seeds of 'Coker 68-15' wheat and 'Maton' rye were immersed for 5 min in acetone solutions of oxamyl, carbofuran, or phenamiphos containing 0, 0.25, 0.5, 1.25, 2.5, or 5.0% (w/v) nematicide; after drying, seeds were planted in pots containing 500 gm of sandy loam naturally infested with Hoplolaimus galeatus and Tylenchorhynchus claytoni. In sterilized soil, only the 5% concentrations of all nematicides were toxic to rye, whereas both the 2.5 and 5% concentrations were damaging to wheat. Phenamiphos was generally the most phytotoxic compound. Numbers of T. claytoni in soil declined sharply in response to seed treatment with all nematicides. In soil planted with wheat, numbers were reduced 80% by the 1.25% treatment; little additional control was shown with higher concentrations. Soil with rye showed a 40-60% reduction in numbers of T. claytoni with the 1.25% solutions and little change at higher concentrations. Hoplolaimus galeatus developed only in pots with rye; root populations were suppressed (30-50%) by treatment with 1.25% or higher concentrations of all nematicides. PMID- 19305617 TI - Seed treatment as a means of preventing nematode damage to crop plants. AB - A procedure for treating crop seeds with aqueous solutions of the systemic nematicide oxamyl is described. Seedlings from treated seeds were more resistant to attack by parasitic nematodes. Leachate from treated seeds reduced the number of free-living nematodes in the surrounding soil. PMID- 19305618 TI - Japanese hollies: intolerant hosts of Meloidogyne arenaria in microplots. AB - Japanese hollies were itttolerant of Meloidogyne arenaria in field microplot experiments. Ilex crenata var. rotundifolia was relatively more tolerant than I. crenata var. convexa or I. crenata var. helleri. When M. arenaria was added at various itfitial population densities to soil containing plants of "Helleri," "Convexa," and "Rotundifolia," respectively, 91, 75, and 25% were killed by the end of the third growing season. No control plants died during the same period. Initial numbers of M. arenaria larvae and eggs were the only population densities that were correlated (negatively), regardless of cultivar, with plant growth over the three growing seasons. A linear relation was found for initial density of M. arenaria and growth of I. crenata rotundifolia. Increasing nematode density by 10 fold suppressed the growth of this cultivar by 23%. PMID- 19305619 TI - Cryofracturing for scanning electron microscope observations of internal structures of nematodes. AB - Nematodes were prepared for scanning electron microscopy by cryofracturing in ethanol and then by critical-point drying in carbon dioxide. Cross sections of Caenorhabditis briggsae and Xiphinema americanum showed the arrangement of the intestine, ovaries, muscle cells, and some layers of the cuticle. The technique is complementary to transmission electron microscopy and facilitates the interpretation of results from thin sections. PMID- 19305620 TI - Howardula dominicki n. sp. infesting the tobacco flea bettle in North Carolina. AB - Howardula dominicki n. sp. is described from specimens collected from the tobacco flea beetle, Epitrix hirtipennis (Melsheimer), at Oxford, North Carolina , and is distinguished from other members of the genus . Parasitism by H. dominicki sterilized female flea beetles and often led to the death of larvae. PMID- 19305621 TI - Effects of host size and parasite burden on sex ratio in the mosquito parasite Octomyomermis muspratti. AB - The ratio of Octomyomermis muspratti to the host mosquito at the time of exposure had little effect on the ratio of male to female parasites that resulted. However, the ratio of males to females increased as the number of parasites/host increased. Hosts with a single nematode produced fewer than 1% males in comparison with hosts with 8 parasites which produced about 40% males; hosts with 10 or more nematodes generally produced more male than female nematodes. Males of O. muspratti usually emerged before females because of the earlier death of multiply-infected mosquitoes. The size of the host at the time of invasion bad no significant influence on nematode sex ratios. Since mating is apparently necessary for reproduction in O. muspratti, the low male to female ratios that occur will be important in developing successful mass production techniques. PMID- 19305622 TI - Development of the DD-136 strain of Neoaplectana carpocapsae at constant temperatures. AB - The development of the DD-136 strain of Neoaplectana carpocapsae was studied on three food sources at 10, 15, 20, 25, 30, 33, 35, and 37 C. No growth occurred at 10 or above 33 C. At 15, 20. and 25 C, growth and reproduction occurred. The most favorable growth occurred at 25 C. At 30 C, N. carpocapsae developed to adults but did not reproduce. Key Words: temperature-growth effects, DD-136 strain. PMID- 19305623 TI - Interaction of plastic, hay and grass mulches, and metam-sodium on control of Pratylenchus penetrans in tomatoes. PMID- 19305624 TI - Reactions of sorghum-sudangrass hybrids and pearl millet to three species of Meloidogyne. PMID- 19305625 TI - Standards of excellence and open questions in cancer biomarker research: an informatics perspective. PMID- 19305626 TI - Sarah Amy Hill. PMID- 19305627 TI - Bias, randomization, and ovarian proteomic data: a reply to "producers and consumers". AB - Proteomic patterns derived from mass spectrometry have recently been put forth as potential biomarkers for the early diagnosis of cancer. This approach has generated much excitement, particularly as initial results reported on SELDI profiling of serum suggested that near perfect sensitivity and specificity could be achieved in diagnosing ovarian cancer. However, more recent reports have suggested that much of the observed structure could be due to the presence of experimental bias. A rebuttal to the findings of bias, subtitled "Producers and Consumers", lists several objections. In this paper, we attempt to address these objections. While we continue to find evidence of experimental bias, we emphasize that the problems found are associated with experimental design and processing, and can be avoided in future studies. PMID- 19305628 TI - Proteomics: analysis of spectral data. AB - The goal of disease-related proteogenomic research is a complete description of the unfolding of the disease process from its origin to its cure. With a properly selected patient cohort and correctly collected, processed, analyzed data, large scale proteomic spectra may be able to provide much of the information necessary for achieving this goal. Protein spectra, which are one way of representing protein expression, can be extremely useful clinically since they can be generated from blood rather than from diseased tissue. At the same time, the analysis of circulating proteins in blood presents unique challenges because of their heterogeneity, blood contains a large number of different abundance proteins generated by tissues throughout the body. Another challenge is that protein spectra are massively parallel information. One can choose to perform top down analysis, where the entire spectra is examined and candidate peaks are selected for further assessment. Or one can choose a bottom-up analysis, where, via hypothesis testing, individual proteins are identified in the spectra and related to the disease process. Each approach has advantages and disadvantages that must be understood if protein spectral data are to be properly analyzed. With either approach, several levels of information must be integrated into a predictive model. This model will allow us to detect disease and it will allow us to discover therapeutic interventions that reduce the risk of disease in at-risk individuals and effectively treat newly diagnosed disease. PMID- 19305629 TI - Design of early validation trials of biomarkers. AB - The design of early-phase studies of putative screening markers in clinical populations is discussed. Biological, epidemiological, statistical and computational issues all affect the design of early-phase studies of these markers, but there are frequently little or no data in hand to facilitate the design. Early-phase studies must be designed as part of a development program, considering the final use of the marker, directly informing the decision to made at the study's conclusion. Therefore, they should test for sensitivity and specificity that would be minimally acceptable to proceed to the next stage of development. Designing such trials requires explicit assumptions about prevalence and false positive and negative costs in the ultimate target population. Early discussion of these issues strengthens the development process, since enthusiasm for developing technologies is balanced by realism about the requirements of a valid population screen. Receiver operating characteristic (ROC) curves, which are useful descriptive tools, may be misleading when evaluating tests in low prevalence populations, because they emphasize the relationship between specificity and sensitivity in the range of specificity likely to be too low to be useful in mass screening applications. PMID- 19305630 TI - Characterising phase variations in MALDI-TOF data and correcting them by peak alignment. AB - The use of MALDI-TOF mass spectrometry as a means of analyzing the proteome has been evaluated extensively in recent years. One of the limitations of this technique that has impeded the development of robust data analysis algorithms is the variability in the location of protein ion signals along the x-axis. We studied technical variations of MALDI-TOF measurements in the context of proteomics profiling. By acquiring a benchmark data set with five replicates, we estimated 76% to 85% of the total variance is due to phase variation. We devised a lobster plot, so named because of the resemblance to a lobster claw, to help detect the phase variation in replicates. We also investigated a peak alignment algorithm to remove the phase variation. This operation is analogous to the normalization step in microarray data analysis. Only after this critical step can features of biological interest be clearly revealed. With the help of principal component analysis, we demonstrated that after peak alignment, the differences among replicates are reduced. We compared this approach to peak alignment with a model-based calibration approach in which there was known information about peaks in common among all spectra. Finally, we examined the potential value at each point in an analysis pipeline of having a set of methods available that includes parametric, semiparametric and nonparametric methods; among such methods are those that benefit from the use of prior information. PMID- 19305631 TI - Understanding the characteristics of mass spectrometry data through the use of simulation. AB - BACKGROUND: Mass spectrometry is actively being used to discover disease-related proteomic patterns in complex mixtures of proteins derived from tissue samples or from easily obtained biological fluids. The potential importance of these clinical applications has made the development of better methods for processing and analyzing the data an active area of research. It is, however, difficult to determine which methods are better without knowing the true biochemical composition of the samples used in the experiments. METHODS: We developed a mathematical model based on the physics of a simple MALDI-TOF mass spectrometer with time-lag focusing. Using this model, we implemented a statistical simulation of mass spectra. We used the simulation to explore some of the basicoperating characteristics of MALDI or SELDI instruments. RESULTS: The simulation reproduced several characteristics of actual instruments. We found that the relative mass error is affected by the time discretization of the detector (about 0.01%) and the spread of initial velocities (about 0.1%). The accuracy of calibration based on external standards decays rapidly outside the range spanned by the calibrants. Natural isotope distributions play a major role inbroadening peaks associated with individual proteins. The area of a peak is a more accurate measure of its size than the height. CONCLUSIONS: The model described here is capable of simulating realistic mass spectra. The simulation should become a useful tool forgenerating spectra where the true inputs are known, allowing researchers to evaluate the performance of new methods for processing and analyzing mass spectra. AVAILABILITY: http://bioinformatics.mdanderson.org/cromwell.html. PMID- 19305632 TI - Assessing the statistical significance of the achieved classification error of classifiers constructed using serum peptide profiles, and a prescription for random sampling repeated studies for massive high-throughput genomic and proteomic studies. AB - Peptide profiles generated using SELDI/MALDI time of flight mass spectrometry provide a promising source of patient-specific information with high potential impact on the early detection and classification of cancer and other diseases. The new profiling technology comes, however, with numerous challenges and concerns. Particularly important are concerns of reproducibility of classification results and their significance. In this work we describe a computational validation framework, called PACE (Permutation-Achieved Classification Error), that lets us assess, for a given classification model, the significance of the Achieved Classification Error (ACE) on the profile data. The framework compares the performance statistic of the classifier on true data samples and checks if these are consistent with the behavior of the classifier on the same data with randomly reassigned class labels. A statistically significant ACE increases our belief that a discriminative signal was found in the data. The advantage of PACE analysis is that it can be easily combined with any classification model and is relatively easy to interpret. PACE analysis does not protect researchers against confounding in the experimental design, or other sources of systematic or random error. We use PACE analysis to assess significance of classification results we have achieved on a number of published data sets. The results show that many of these datasets indeed possess a signal that leads to a statistically significant ACE. PMID- 19305633 TI - Novel approaches to smoothing and comparing SELDI TOF spectra. AB - BACKGROUND: Most published literature using SELDI-TOF has used traditional techniques in Spectral Analysis such as Fourier transforms and wavelets for denoising. Most of these publications also compare spectra using their most prominent feature, i.e, peaks or local maximums. METHODS: The maximum intensity value within each window of differentiable m/z values was used to represent the intensity level in that window. We also calculated the 'Area under the Curve' (AUC) spanned by each window. RESULTS: Keeping everything else constant, such as pre-processing of the data and the classifier used, the AUC performed much better as a metric of comparison than the peaks in two out of three data sets. In the third data set both metrics performed equivalently. CONCLUSIONS: This study shows that the feature used to compare spectra can have an impact on the results of a study attempting to identify biomarkers using SELDI TOF data. PMID- 19305634 TI - the need for review and understanding of SELDI/MALDI mass spectroscopy data prior to analysis. AB - Multiple studies have reported that surface enhanced laser desorption/ionization time of flight mass spectroscopy (SELDI-TOF-MS) is useful in the early detection of disease based on the analysis of bodily fluids. Use of any multiplex mass spectroscopy based approach as in the analysis of bodily fluids to detect disease must be analyzed with great care due to the susceptibility of multiplex and mass spectroscopy methods to biases introduced via experimental design, patient samples, and/or methodology. Specific biases include those related to experimental design, patients, samples, protein chips, chip reader and spectral analysis. Contributions to biases based on patients include demographics (e.g., age, race, ethnicity, sex), homeostasis (e.g., fasting, medications, stress, time of sampling), and site of analysis (hospital, clinic, other). Biases in samples include conditions of sampling (type of sample container, time of processing, time to storage), conditions of storage, (time and temperature of storage), and prior sample manipulation (freeze thaw cycles). Also, there are many potential biases in methodology which can be avoided by careful experimental design including ensuring that cases and controls are analyzed randomly. All the above forms of biases affect any system based on analyzing multiple analytes and especially all mass spectroscopy based methods, not just SELDI-TOF-MS. Also, all current mass spectroscopy systems have relatively low sensitivity compared with immunoassays (e.g., ELISA). There are several problems which may be unique to the SELDI-TOF-MS system marketed by Ciphergen((r)). Of these, the most important is a relatively low resolution (+/-0.2%) of the bundled mass spectrometer which may cause problems with analysis of data. Foremost, this low resolution results in difficulties in determining what constitutes a "peak" if a peak matching approach is used in analysis. Also, once peaks are selected, the peaks may represent multiple proteins. In addition, because peaks may vary slightly in location due to instrumental drift, long term identification of the same peaks may prove to be a challenge. Finally, the Ciphergen((r)) system has some "noise" of the baseline which results from the accumulation of charge in the detector system. Thus, we must be very aware of the factors that may affect the use of proteomics in the early detection of disease, in determining aggressive subsets of cancers, in risk assessment and in monitoring the effectiveness of novel therapies. PMID- 19305635 TI - Impact of freeze-thaw cycles and storage time on plasma samples used in mass spectrometry based biomarker discovery projects. AB - Mass spectrometry approaches to biomarker discovery in human fluids have received a great deal of attention in recent years. While mass spectrometry instrumentation and analysis approaches have been widely investigated, little attention has been paid to how sample handling can impact the plasma proteome and therefore influence biomarker discovery. We have investigated the effects of two main aspects of sample handling on MALDI-TOF data: repeated freeze-thaw cycles and the effects of long-term storage of plasma at -70 degrees C. Repeated freeze thaw cycles resulted in a trend towards increasing changes in peak intensity, particularly after two thaws. However, a 4-year difference in long-term storage appears to have minimal effect on protein in plasma as no differences in peak number, mass distribution, or coefficient of variation were found between samples. Therefore, limiting freeze/thaw cycles seems more important to maintaining the integrity of the plasma proteome than degradation caused by long term storage at -70 degrees C. PMID- 19305636 TI - Preliminary evaluation of the speech perception assessment and training system (SPATS) with hearing-aid and cochlear-implant users. PMID- 19305637 TI - Development of a measure of sense of community for individuals with serious mental illness residing in community settings. AB - The psychological sense of community is one of the most commonly investigated constructs in community psychology. Sense of community may be particularly important for individuals with serious mental illness (SMI) because they often face societal barriers to participation in community living, including stigma and discrimination. To date, no published studies have investigated the psychometric qualities of sense of community measures among individuals with SMI. The current study tested a series of confirmatory factor analyses using the Brief Sense of Community Index (BSCI, Long & Perkins, 2003) in a sample of 416 persons with SMI living in community settings to suggest a model of sense of community for individuals with SMI and other disabilities. The resulting scale, the Brief Sense of Community Index- Disability (BSCI-D), demonstrated good model fit and construct validity. Implications are discussed for how this scale may be used in research investigating community integration and adaptive functioning in community settings. PMID- 19305638 TI - Learning to Move. AB - Locomotion-moving the body from place to place-is one of infants' greatest achievements. In addition to conquering gravity, infants must cope with variable and novel constraints on balance and propulsion. At the same time that they are learning to move, changes in infants' bodies, skills, and environments change the biomechanical constraints on movement. Recent work highlights both flexibility and specificity in infants' responses to novel and variable situations, demonstrating that infants are learning to learn as they master locomotion. Within sitting, crawling, cruising, and walking postures, experienced infants adapt their locomotor responses to the current biomechanical constraints on movement. However, what infants have learned about coping with variability and novelty in earlier-developing postures does not transfer to later-developing postures. PMID- 19305639 TI - A genotype-dependent intermediate ECG phenotype in patients with persistent lone atrial fibrillation genotype ECG-phenotype correlation in atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) is heterogeneous at the clinical and molecular levels. Association studies have reported that common single-nucleotide polymorphisms in KCNE1 and SCN5A may predispose to AF. In this study, we tested the hypothesis that specific AF-associated genotypes confer variation on the appearance of AF assessed by analysis of fibrillatory rate of the atria. METHODS AND RESULTS: Twenty-six nonrelated patients (21 males, mean age 55+/-12 years) with persistent lone AF (median AF duration 5 weeks) not taking class I or III antiarrhythmic drugs were studied. Fibrillatory rate was obtained by spatiotemporal QRST cancellation and time-frequency analysis of the index surface ECG. Genotypes at the AF-associated loci in KCNE1 (S38G) and SCN5A (H558R) were determined by direct DNA sequencing. The atrial fibrillatory rate was 418+/-50 fibrillations per minute (range, 336 to 521) in the study cohort. Carriers of the 38GG KCNE1 genotype (n=13) had significantly lower fibrillatory rates (392+/-36 versus 443+/-49 fibrillations per minute, P0.006) compared to those with GS or SS genotype (n=13). Six patients (23%) with fibrillatory rates >450 fibrillations per minute, all had either the GS or SS genotype (Chi2 P0.008). In contrast, both the heterozygeous and homozygeous SCN5A H558R polymorphism had no effect on fibrillatory rate. There were no significant associations between fibrillatory rate and clinical (age, gender, AF duration, drug treatment) or echocardiographic (left atrial diameter, left ventricular ejection fraction) variables. In multivariable regression analysis, the KCNE1 S38G genotype (SS/GS coded 0, GG coded 1) was the only independent predictor of fibrillatory rate (beta = -0.437, P = 0.006) with a SE of the estimate of 44 fibrillations per minute. CONCLUSIONS: This study suggests that atrial fibrillatory rate obtained from the surface ECG is at least in part determined by KCNE1 (S38G) genotype, implying that this variant exerts functional effects on atrial electrophysiology. This intermediate ECG phenotype may be useful for elaborating genetic influences on AF mechanisms and identifying subsets of patients for variability in AF susceptibility or response to therapies. PMID- 19305640 TI - Low-volume, high-throughput sandwich immunoassays for profiling plasma proteins in mice: identification of early-stage systemic inflammation in a mouse model of intestinal cancer. AB - Mouse models of human cancers may provide a valuable resource for the discovery of cancer biomarkers. We have developed a practical strategy for profiling specific proteins in mouse plasma using low-volume sandwich-immunoassays. We used this method to profile the levels of 14 different cytokines, acute-phase reactants, and other cancer markers in plasma from a mouse models of intestinal tumors and their wild-type littermates, using as little as 1.5 microliters of diluted plasma per assay. Many of the proteins were significantly and consistently up-regulated in the mutant mice. The mutant mice could be distinguished nearly perfectly from the wild-type mice based on the combined levels of as few as three markers. Many of the proteins were up-regulated even in the mutant mice with few or no tumors, suggesting the presence of a systemic host response at an early stage of cancer development. These results have implications for the study of host responses in mouse models of cancers and demonstrate the value of a new low-volume, high-throughput sandwich-immunoassay method for sensitively profiling protein levels in cancer. PMID- 19305641 TI - PKC and PKA phosphorylation affect the subcellular localization of claudin-1 in melanoma cells. AB - Cytoplasmic expression of claudin-1 in metastatic melanoma cells correlates to increased migration, and increased secretion of MMP-2 in a PKC dependent manner, whereas claudin-1 nuclear expression is found in benign nevi. Melanoma cells were transfected with a vector expressing CLDN-1 fused to a nuclear localization signal (NLS). Despite significant nuclear localization of claudin-1, there was still transport of claudin-1 to the cytoplasm. Phorbol ester treatment of cells transfected with NLS-claudin-1 resulted in an exclusion of claudin-1 from the nucleus, despite the NLS. To ascertain whether PKC or PKA were involved in this translocation, we mutated the putative phosphorylation sites within the protein. We found that mutating the PKC phosphorylation sites to mimic a non phosphorylated state did not cause a shift of claudin-1 to the nucleus of the cells, but mutating the PKA sites did. Mutations of either site to mimic constitutive phosphorylation resulted in cytoplasmic claudin-1 expression. Stable claudin-1 transfectants containing non-phosphorylatable PKA sites exhibited decreased motility. These data imply that subcellular localization of claudin-1 can be controlled by phosphorylation, dicating effects on metastatic capacity. PMID- 19305642 TI - In vivo evidence of hepato- and reno-protective effect of garlic oil against sodium nitrite-induced oxidative stress. AB - Sodium nitrite (NaNO2), a food color fixative and preservative, contributes to carcinogenesis. We investigated the protective role of garlic oil against NaNO2 induced abnormalities in metabolic biochemical parameters and oxidative status in male albino rats. NaNO2 treatment for a period of three months induced a significant increase in serum levels of glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin, urea and creatinine as well as hepatic AST and ALT. However, significant decrease was recorded in liver ALP activity, glycogen content, and renal urea and creatinine levels. In parallel, a significant increase in lipid peroxidation, and a decrease in glutathione content and catalase activity were observed in the liver and the kidney. However, garlic oil supplementation showed a remarkable amelioration of these abnormalities. Our data indicate that garlic is a phytoantioxidant with powerful chemopreventive properties against chemically induced oxidative stress. PMID- 19305643 TI - The role of SRC-1 in murine prostate cancinogenesis is nonessential due to a possible compensation of SRC-3/AIB1 overexpression. AB - The androgen and androgen receptor (AR)-regulated gene expression plays important roles in normal prostate and prostate cancer development, and AR transcriptional control of genes is mediated by transcriptional coactivators, including the three members of the steroid receptor coactivator (SRC) family, SRC-1 (NCOA1), SRC-2 (TIF2/GRIP1/NCOA2) and SRC-3 (AIB1, ACTR/RAC3/NCOA3). SRC-1 and SRC-3 are overexpressed in multiple human endocrine cancers and knockdown of either one of them in prostate cancer cell lines impedes cellular proliferation. Knockout of SRC-3 in mice suppresses the progression of spontaneous prostate carcinogenesis. In this study, we investigated SRC-1 contribution to prostate cancer in vivo by deleting the SRC-1 gene in TRAMP mice, which contain the probasin promoter-driven SV40 T/t antigen transgene. In assessing tumor mass of mice at various ages, we found that initiation and progression of prostate cancer induced by SV40 T/t antigens were unaltered in SRC-1(-/-) mice versus WT mice. Primary tumor histology and metastasis to distant lymph nodes were also similar in these mice at all time points assessed. These results demonstrate that the role of SRC-1 in mouse prostate carcinogenesis is nonessential and different from the essential contribution of SRC-3 that is required for prostate cancer progression and metastasis in mice. Interestingly, we observed that during prostate tumorigenesis SRC-1 expression was relatively constant, while SRC-3 expression was significantly elevated. Therefore, the loss of SRC-1 function may be compensated by SRC-3 overexpression during prostate tumorigenesis in SRC-1(-/-) mice. PMID- 19305644 TI - Corticosterone alters AMPAR mobility and facilitates bidirectional synaptic plasticity. AB - BACKGROUND: The stress hormone corticosterone has the ability both to enhance and suppress synaptic plasticity and learning and memory processes. However, until today there is very little known about the molecular mechanism that underlies the bidirectional effects of stress and corticosteroid hormones on synaptic efficacy and learning and memory processes. In this study we investigate the relationship between corticosterone and AMPA receptors which play a critical role in activity dependent plasticity and hippocampal-dependent learning. METHODOLOGY/PRINCIPAL FINDINGS: Using immunocytochemistry and live cell imaging techniques we show that corticosterone selectively increases surface expression of the AMPAR subunit GluR2 in primary hippocampal cultures via a glucocorticoid receptor and protein synthesis dependent mechanism. In agreement, we report that corticosterone also dramatically increases the fraction of surface expressed GluR2 that undergo lateral diffusion. Furthermore, our data indicate that corticosterone facilitates NMDAR-invoked endocytosis of both synaptic and extra-synaptic GluR2 under conditions that weaken synaptic transmission. CONCLUSION/SIGNIFICANCE: Our results reveal that corticosterone increases mobile GluR2 containing AMPARs. The enhanced lateral diffusion properties can both facilitate the recruitment of AMPARs but under appropriate conditions facilitate the loss of synaptic AMPARs (LTD). These actions may underlie both the facilitating and suppressive effects of corticosteroid hormones on synaptic plasticity and learning and memory and suggest that these hormones accentuate synaptic efficacy. PMID- 19305645 TI - Is American health care uniquely inefficient? PMID- 19305647 TI - Synthesis of Amine-stabilized Aqueous Colloidal Iron Oxide Nanoparticles. AB - We demonstrate a simple one-step process for the synthesis of iron oxide nanoparticle aqueous colloids using the multifunctional molecule, dodecylamine (DDA), that electrostatically complexes with aqueous iron ions (one precursor Fe(2+) from FeCl(2)), reduces them, and subsequently caps the nanoparticles. The iron oxide particles thus synthesized are of the face-centered cubic (FCC) phase with high degree of monodispersity with appropriate concentration of amine capping molecular layer. The aqueous magnetic nanocrystalline colloids were characterized by TEM, XRD, XPS, TGA/DTA and FTIR spectroscopy techniques. The relaxivity, stability, and hydrodynamic size of the nanoparticles were investigated for potential application in magnetic resonance imaging (MRI). The magnetic properties were also studied by using a superconducting quantum interference device (SQUID) magnetometer at room temperature. We believe that such simple one-step synthesis of biocompatible aqueous nanomagnetic colloids will have viable applications in biomedical imaging, diagnostics and therapeutics. PMID- 19305648 TI - Researchers' role in policy decision-making: purveyors of evidence, purveyors of ideas? PMID- 19305646 TI - Potential Benefits of Peroxynitrite. AB - Peroxynitrite (PN) is generated by the reaction of nitric oxide (NO) and superoxide in one of the most rapid reactions in biology. Studies have reported that PN is a cytotoxic molecule that contributes to vascular injury in a number of disease states. However, it has become apparent that PN has beneficial effects including vasodilation, inhibition of platelet aggregation, inhibition of inflammatory cell adhesion, and protection against ischemia/reperfusion injury in the heart. It is our hypothesis that PN may serve to inactivate superoxide and prolong the actions of NO in the circulation. This manuscript reviews the beneficial effects of PN in the cardiovascular system. PMID- 19305649 TI - Kafka, New Orleans, the OARs and the KT boundary: reverse the flow of Kafka's fable of the Imperial Message and you have the problem of Knowledge Transfer--the message is not getting through. AB - The dying emperor in Kafka's tale has sent you a message of great importance, entrusted to his strongest herald. But the throne room, the palace, the city are so huge and so crowded with people that the message can never reach you. Reverse the tale and you have the problem of Knowledge Transfer (KT)--brutally illustrated by the fate of New Orleans. Important information, of unquestioned validity, soundly based in research and easy to understand, simply does not get through to practice. A recent evaluation of the Ottawa Ankle Rules makes the point all too clearly. Indeed, the KT problem is worse than Kafka's. Economically motivated people and organizations actively distort the messages--and try to trip the herald--while grossly oversimplified frameworks of understanding include no language in which the messages can be expressed. "More research" is not the answer. PMID- 19305650 TI - Working within and beyond the Cochrane Collaboration to make systematic reviews more useful to healthcare managers and policy makers. AB - Participants in the Cochrane Collaboration conduct and periodically update systematic reviews that address the question, "What works?" for healthcare interventions. The Cochrane Library makes available quality-appraised systematic reviews that address this question. No coordinated effort has been undertaken to conduct and periodically update systematic reviews that address the other types of questions asked by healthcare managers and policy makers, to adapt existing reviews to highlight decision-relevant information (including the factors that may affect assessments of a review's local applicability) or to facilitate their retrieval through a "one-stop shopping" portal. Researchers interested in evaluating new methodological developments, health services and policy researchers interested in conducting and adapting systematic reviews, and research funders all have a role to play in making systematic reviews more useful for healthcare managers and policy makers. PMID- 19305651 TI - Reframing evidence synthesis as rhetorical action in the policy making drama. AB - This paper presents a novel conceptualization of policy making as social drama. The selection and presentation of evidence for policy making, including the choice of which questions to ask, which evidence to compile in a synthesis and which syntheses to bring to the policy making table, should be considered as moves in a rhetorical argumentation game and not as the harvesting of objective facts to be fed into a logical decision-making sequence. Viewing policy making as argument does not mean it is beyond rationality--merely that we must redefine rationality to include not only logical inference and probabilistic reasoning, but also the consideration of plausibility by a reasonable audience. We need better evidence, but we also urgently need better awareness by policy makers of the language games on which their work depends. PMID- 19305652 TI - Informing policy making and management in healthcare: the place for synthesis. AB - Research synthesis has an important role supporting the transfer of knowledge between researchers and healthcare decision-makers. But if our goal is to make knowledge more useable and context specific, then extending the scope of systematic reviews or producing syntheses with policy makers and managers may be insufficient. Dialogues, partnerships and reinterpretations of evidence in context will help us achieve this goal. PMID- 19305653 TI - A decision-maker's perspective on Lavis and Lomas. AB - Advancement in research synthesis, so well articulated and advocated by Lomas (2005) and Lavis et al. (2005), is a necessary but not sufficient development for the systematized use of research in managerial practice. Although significant progress in the use of evidence-based approaches can be witnessed, enriching efforts need to progress within both the management and research communities. Contextualized by direct experience with harm reduction approaches to population health improvement, this commentary offers propositions concerning the nature of the researcher/decision-maker interchange, arguing for a pronounced strengthening of involvement and activity at all levels in the service delivery system. PMID- 19305654 TI - Commentary: whose views count in evidence synthesis? And when do they count? AB - Four recent pieces in Healthcare Policy reveal some disagreement on when and how to involve decision-makers in the process of evidence synthesis. This commentary proposes varying roles for researchers versus managers or policy makers at each of three different stages of synthesis and at the actual point of decision. It also raises the issue of how poorly current processes accommodate the broader conception of evidence held by most managers and policy makers. PMID- 19305655 TI - Moving forward on both systematic reviews and deliberative processes. AB - Systematic reviews are increasingly seen as helpful "knowledge support" for managers and policy makers, and deliberative processes are starting to be seen as promising, locally contextualized "decision support." Increases to the flow of systematic reviews should be complemented by efforts to facilitate the retrieval, and adapt the presentation, of the available stock of systematic reviews. Research and other evidence should be combined in transparent ways to facilitate cross-context learning. The challenge for managers and policy makers in moving forward will be to avoid the confusion that comes from the branding of both systematic reviews and deliberative processes. PMID- 19305656 TI - Bariatric surgery in Canada: obesity rates for Canadian adults are much higher today than in the past; however, rates of bariatric surgery, a treatment for high risk severely obese individuals, have not risen in parallel. AB - Obesity rates for Canadian adults are much higher today than in the past, raising questions about how to achieve healthy weights and mitigate the associated health risks. While not a solution at the population level, bariatric surgery may be a treatment option for a relatively small proportion of obese individuals. In Canada, unlike in the United States, no consistent trend was evident in the use of this surgery between 1996-97 and 2003-04 across the five provinces for which comparable data were available. In 2003-04, bariatric surgeries were performed predominantly for women (87%); the average length of stay in hospital was 5 days; and 1.4% of patients were readmitted to hospital within 7 days of their discharge after surgery. PMID- 19305657 TI - Reflections on conversations with Robert Bell and Michael Guerriere: what is relevant research? AB - Two decision-makers from the acute-care sector weigh in on the issue of relevant research. Between the two of them they look for patient-defined research, evidence to support the conclusions, information that can lead to interventions designed to improve quality and outcomes and defined control mechanisms to properly identify the practices that improved the system. Three examples are cited and discussed. The context is set by comments from one of Canada's leading researchers and the use of research from one of this decade's most lauded system turnarounds. PMID- 19305658 TI - Knowledge translation to advance the nurse practitioner role in British Columbia: researchers and decision-makers conduct policy-relevant research to guide legislative and regulatory development and the design of a nurse practitioner education program. AB - This project brought together a team of researchers and decision-makers to conduct policy-relevant research to support the introduction of advanced nursing practice roles in British Columbia. All team members, including decision-makers, were actively involved in the conceptualization, design, data collection, analysis and interpretation of the study. This level of engagement, coupled with ongoing knowledge translation (KT) activities, led to the implementation by stakeholders of a majority of the study's recommendations. The results have since been used to guide legislative and regulatory development and to design a nurse practitioner education program. PMID- 19305659 TI - Interactions: trade policy and healthcare reform after Chaoulli v. Quebec: is it time for Canada to acknowledge the fragile boundary between health and trade policies and strengthen the separation between private and public health insurance? AB - The insulation of Canada's healthcare system from trade treaty obligations is crucial to the legitimacy of Canada's trade policy. Legal analysis has suggested, however, that competitive and for-profit delivery of the kind contemplated by the Kirby Report and some provinces may make healthcare more vulnerable to challenges under NAFTA and GATS. The Government of Canada has tried to counter this interpretation by stressing the importance of public financing as the principal criterion for exemption of healthcare from trade treaties, but now the potential for private financing of essential medical services indicated by the Supreme Court's decision in Chaoulli v. Quebec has made that line of argument look risky as well. It is apparent that Canada failed to anticipate the possible interactions of domestic, international and constitutional law when it made commitments in the area of private health insurance at the WTO in 1997. Accordingly, the time has come to acknowledge the fragility of the boundary between health and trade policies, to take the risks and costs associated with trade treaty obligations fully into account when undertaking healthcare reform and to strengthen the separation between private and public health insurance. PMID- 19305660 TI - Experiencing difficulties accessing first-contact health services in Canada: Canadians without regular doctors and recent immigrants have difficulties accessing first-contact healthcare services. Reports of difficulties in accessing care vary by age, sex and region. AB - In this study, we identify the significant factors associated with having difficulties accessing first-contact healthcare services. Population-based data from two national health surveys, the Health Services Access Survey and the Canadian Community Health Survey, were used to identify respondents who required first-contact services for themselves or for a family member during 2003. Fifteen percent of Canadians reported difficulty accessing routine care, and 23% reported difficulties with immediate care. Physician/service availability was the chief reason cited for difficulties accessing routine care, while for urgent care, it was long wait times. Women, younger respondents and residents of eastern Canada and Quebec were consistently more likely to report difficulties accessing both types of these first-contact services, whereas less educated Canadians were less likely to report problems. Canadians without a regular family doctor were more than twice as likely to report difficulties accessing routine care compared to those who had a regular doctor. New immigrants were almost two and a half times more likely to report difficulties accessing immediate care than were Canadian born respondents. Household income was not associated with difficulties accessing either type of care. The relatively low level of reporting of difficulties by older and less educated Canadians may be related, in part, to more modest expectations about the healthcare system. PMID- 19305661 TI - Inclusivity and dementia: health services planning with individuals with dementia: effective inclusion requires action at multiple levels by individuals with dementia, care partners, service providers and funding organizations. AB - Historically, Alzheimer societies have identified the care partners of persons with dementia as their central clients. This focus is broadening to include the person with the disease, as well. This paper presents the results of a Canadian research study addressing organizational considerations related to effective inclusion of persons with dementia in planning and decision-making about health services and programs. Our findings suggest that effective inclusion requires action at multiple levels by individuals with dementia, care partners and friends; service organizations and providers; and funding organizations. Additional research is needed to explore the applicability of these findings to other organizations in different localities and to examine emergent themes further. Of these, one that has received little attention to date concerns the potential risks associated with effective inclusion. PMID- 19305662 TI - The cost-effectiveness of expanding intensive behavioural intervention to all autistic children in Ontario: in the past year, several court cases have been brought against provincial governments to increase funding for Intensive Behavioural Intervention (IBI). This economic evaluation examines the costs and consequences of expanding an IBI program. AB - Intensive Behavioural Intervention (IBI) describes behavioural therapies provided to autistic children to overcome intellectual and functional disabilities. The high cost of IBI has caused concern regarding access, and recently, several court cases have been brought against provincial governments to increase funding for this intervention. This economic evaluation assessed the costs and consequences of expanding an IBI program from current coverage for one-third of children to all autistic children aged two to five in Ontario, Canada. Data on the hours and costs of IBI, and costs of educational and respite services, were obtained from the government. Data on program efficacy were obtained from the literature. These data were modelled to determine the incremental cost savings and gains in dependency-free life years. Total savings from expansion of the current program were $45,133,011 in 2003 Canadian dollars. Under our model parameters, expansion of IBI to all eligible children represents a cost-saving policy whereby total costs of care for autistic individuals are lower and gains in dependency-free life years are higher. Sensitivity analyses carried out to address uncertainty and lack of good evidence for IBI efficacy and appropriate discount rates yielded mixed results: expansion was not cost saving with discount rates of 5% or higher and with lower IBI efficacy beyond a certain threshold. Further research on the efficacy of IBI is recommended. PMID- 19305663 TI - Waiting time for radiation therapy in breast cancer patients in Quebec from 1992 to 1998: a study of surgically treated breast cancer patients in Quebec documents and helps to explain increased waiting times for radiation therapy. AB - BACKGROUND: This study was conducted among surgically treated breast cancer patients in Quebec to determine waiting time between surgery and post-operative radiation therapy and factors influencing it. METHODS: Records of fee-for-service claims and hospitalizations were obtained for all women who, between 1992 and 1998, underwent an invasive procedure for non-metastatic breast cancer. Waiting time was defined as the time between either the last surgical procedure or the last cycle of chemotherapy and the initiation of radiation therapy. Hierarchical linear regression models were used to identify predictors of waiting time. RESULTS: Over seven years, 29,072 episodes of breast cancer treatments were identified, of which 17,684 included radiation therapy. The number of cases increased by 5.5% per year, but concurrent broadening of indications for radiation therapy led to an increase in the number of breast cancer patients receiving radiation therapy of 9% per year. In hierarchical linear modelling, comparing 1998 to 1992, median waiting time increased by 63% (95% confidence interval [CI] 35%-97%) in patients not receiving chemotherapy and by 35% (95% CI 3%-88%) in patients receiving chemotherapy. Other predictors of shorter waiting times were localized cancer stage, breast-conserving surgery, early consultation with a radiation oncologist, having surgery in a centre with a radiation therapy facility, living close to a radiation therapy facility and living in a higher socio-economic area. INTERPRETATION: Using administrative databases to evaluate waiting times is feasible. Explanations of the increased waiting time include increased demand, insufficient resources and changes in the indications for breast-conserving surgery and radiation therapy. PMID- 19305665 TI - The blind men, the elephant and the CT scanner. AB - The parable of the blind men and the elephant suggests that disputes among scholars arise not so much from errors of fact and argument as from differences of perspective--incomplete perceptions, each from a different angle of view, of a more complex reality. The CT scanner offers an analogy, taking two-dimensional images from multiple points of view. The heart of the device is the computational process integrating these images into a three-dimensional view consistent with each. Effective interdisciplinary research requires the institutional equivalent of the CT scanner's integrative capacity. The normal dynamics of university-based research, however, pull instead toward disciplinary solitudes. Our new journal is a step in the right direction, but there is still a distance to go. PMID- 19305666 TI - Complementary and alternative medicine: a rising healthcare issue. AB - More than half of all Canadians use some form of complementary and alternative medicine (CAM) every year. The way CAM is being used, the magnitude of its use and the lack of clarity on standards of evidence make CAM a rising healthcare issue. A recent research priority-setting exercise by the Canadian Interdisciplinary Network for CAM Research (IN-CAM) identified three research priority areas: (1) healthcare delivery and policy research, including (a) exploring if and how CAM should be regulated, (b) defining what constitutes acceptable evidence of safety and efficacy, (c) investigating the organization and delivery of integrative healthcare; (2) methodological research, including exploring how best to assess whole systems of care and how to choose patient-, practitioner- and policy-relevant outcome measures; and (3) knowledge transfer, including formal education strategies, the provision of information and dialogue with those who use information in decision-making. The high use of CAM products and therapies leads to many questions from patients, practitioners and policy makers. The research agenda presented here provides a guide to begin programs of research that will answer these questions. PMID- 19305667 TI - When leaders talk to leaders. AB - Presentations by three Canadian executives who influence healthcare policy and practice were reviewed to identify the kinds of sources these leaders use to draw their conclusions. All three speakers examined policies, practices and outcomes. Presentations were selected to cover activities in three provinces in three different calendar years, to avoid duplication or undue influence of a particular event or release of information. All three speakers drew heavily on data from government sources, especially the Canadian Institute for Health Information (CIHI). PMID- 19305668 TI - Obesity and joint replacement surgery in Canada: findings from the Canadian Joint Replacement Registry (CJRR). AB - Obesity has been strongly implicated as a risk factor for knee osteoarthritis and, in some studies, for osteoarthritis of the hip. Osteoarthritis is the most commonly reported diagnosis for joint replacement patients. In this study, we conducted analyses based on data from the Canadian Joint Replacement Registry (CJRR) to estimate the relationships between overweight and obesity and rates of joint replacement surgery in Canada. Obese persons were over three times as likely and overweight persons were one and a half times more likely to undergo joint replacement surgery, compared to those in the acceptable weight category in 2003-04. This study provides evidence of a clinically relevant association between obesity and joint replacement surgery. PMID- 19305669 TI - Knowledge translation and patient safety: the Canadian Adverse Events Study. AB - The Canadian Adverse Events Study was the first national study of adverse events in Canadian hospitals. Learning from the controversy surrounding similar studies in other countries, the team engaged in extensive knowledge translation activities throughout the life of the project. Using meetings, Web-based communication and other tools, the team successfully prepared most Canadian stakeholders for the study's release, allowing them to develop anticipatory patient safety initiatives. However, upon publication of the study, the policy spotlight quickly shifted to other issues, and the long-term commitment needed to create safer healthcare is still uncertain. PMID- 19305670 TI - Learning from other sites. AB - What should Canadian healthcare researchers pay attention to on the World Wide Web? What might be done to magnify the Canadian footprint on the Internet? This commentary discusses two very successful sites--OpenCourseWare from the Massachusetts Institute of Technology and the Epidemiology Supercourse from the University of Pittsburgh. The Mythbuster series (from the Canadian Health Services Research Foundation) and the Regional Training Centres have Web presences that might benefit from applying the techniques used by OpenCourseWare and the Supercourse. For a relatively small investment, Canadian efforts to improve dissemination and teaching in the health policy/health services research area might reach a larger national and international audience. PMID- 19305671 TI - The public endorses collection of ethnicity information in hospital: implications for routine data capture in Canadian health systems. AB - A telephone survey was conducted in Calgary, Alberta to assess public opinion on collection of ethnicity information in hospitals. Of the 2,799 respondents, 84.8% felt comfortable about recording their ethnicity in hospital charts. This rate held across respondents' age, marital status and ethnic origin. These findings suggest that Canadian health systems should explore the feasibility and ethical suitability of collecting ethnicity data, as this information could contribute to the evaluation and subsequent reduction of ethnic disparities in health and health services access. PMID- 19305672 TI - Where are nurses working? Employment patterns by sub-sector in Ontario, Canada. AB - OBJECTIVE: As care shifts to home and community, have nursing jobs followed? We examined changes in the absolute and relative size of the nursing workforce by sector/sub-sectors in Ontario, Canada. METHODS: All nurses registered with the Ontario College of Nurses over the 11 years from 1993 to 2003 were categorized as Active, Eligible or Not Eligible. Active nurses were then categorized by sector (Hospital, Community, Other) and sub-sector. The analysis was repeated by age group and for registered nurses and registered practical nurses. RESULTS: The decline in Active and Eligible nurses was particularly pronounced for younger workers. Both the absolute number and proportion of nurses working in the hospital sub-sector has dropped. In the community sector, growth was evident in the use of nurses as case managers (in the CCAC sub-sector), community agencies and community mental health (representing a shift from hospital-based workers). However, the steady growth in the number and proportion of nurses working in home care agencies was reversed in 1999, with this sub-sector shedding 19% of its nurses by 2003. CONCLUSION: Despite considerable rhetoric to the contrary, nurses still tend to work within institutions (hospitals and long-term-care facilities). However, compared to their numbers in 1993, there were fewer nurses providing direct patient care in Ontario in both the hospital and community sectors, and a higher proportion of older nurses. PMID- 19305673 TI - Turning 65 in Ontario: the impact of public drug benefit coverage on hospitalizations for acute and chronic disease. AB - Ongoing access to medication is required for the management of many chronic diseases. We sought to examine the health impact of entering the provincial drug benefit plan at age 65 years. We constructed cohorts of same-aged individuals with incident heart failure, with diabetes and from the whole population using administrative data. The hazard of hospitalization at a given age compared to one year younger for ages 63 to 67 years was determined. On turning age 65, persons were more rather than less likely to be hospitalized for diabetes and acute infection when compared to the prior year. Among residents of low-income neighbourhoods these effects were no longer significant, but the risk of hospitalization for heart failure was increased. A similar effect was not observed for the paired cohorts at the transition from age 63 to 64 years. A health benefit of obtaining drug insurance was not observed in this study; however, the relationship may have been confounded by the effects of retirement and other changes occurring at age 65. Moreover, a benefit experienced by only a small proportion of the population may not be apparent in this analysis, which included many individuals unaffected by the policy. PMID- 19305674 TI - Implementation evaluation of an integrated healthcare delivery initiative for diabetic patients. AB - The aim of this in-depth, longitudinal, qualitative case study was to understand and explain the dynamics of implementing a pilot project to deliver integrated healthcare for type 2 diabetic patients. Data gathering included in-depth, face to-face interviews with family physicians, nurses and other healthcare providers, managers and policy makers (n = 25) at various points during the research period, and focus groups (n = 3) with patients. These data were complemented by onsite observations of numerous committee meetings, and analysis of project documentation. Benson's (1975) political economy perspective provided a valuable conceptual framework for tracking the complex dynamics of implementing service integration. Manipulative strategies (continuing medical education, new information technology) did not engage physicians. Of the cooperative strategies attempted (disease management, patient education, community mobilization), only patient education appears to have succeeded: patients acknowledged that project educational activities enabled them to improve self-management of their disease. However, the project's emphasis on patient education effectively increased nurses' bargaining power within the healthcare team, to the detriment of the focus on integrated care. Integrating care is a laborious process that takes time to reach fruition. This one-year pilot project was insufficient for mobilizing health professionals away from fragmented practices toward integrated ones. New resources mostly allowed them to maintain or increase their power positions within the network of care providers. Nevertheless, this initiative raised physicians' awareness and appreciation of the care that other health professionals provided to their patients. PMID- 19305676 TI - High reliability versus high autonomy: Dryden, Murphy and patient safety. AB - Healthcare is not a high-reliability industry. The adverse event rate is on the order of 10(-2); industries such as aviation, nuclear power and railways achieve rates of 10(-5) or better. Increasing awareness of this contrast has made "patient safety" a major topic of concern. High reliability in other industries flows from a combination of "engineered safety," tight regulation ("high-level constraints") and the development of a "culture of safety" that recognizes error as a systemic rather than a personal failure. In medicine, achieving such a combination would involve abandoning deeply embedded and centuries-old traditions of individualism, clinical autonomy and personal responsibility. This will not happen. Watch instead for safety concerns to be diverted into activities that do not threaten core values. PMID- 19305677 TI - Research output of the Canadian pharmaceutical industry: where has all the R&D gone? AB - Although the Canadian pharmaceutical industry claims to be spending about $1 billion per year for research and development (R&D), there is little evidence of research performed, as measured by scientific publications and patent applications. One firm was exceptional; it compared favourably with its parent firm in regard to the number of publications and patents in relation to the R&D budget, demonstrating the feasibility of developing a productive and independent research program in Canada. The perception of low productivity is made worse by the inadequacy of the annual report on R&D prepared by the Patented Medicines Prices Review Board (PMPRB). We recommend a number of changes in the PMPRB's mandate so that its collection of R&D data and subsequent analysis will be more complete. Further financial assistance to the industry should be withheld until accountability is assured and a full assessment of the outcome of its R&D program can be made. PMID- 19305678 TI - Missed opportunity: patients who leave emergency departments without being seen. AB - The rate of patients who visit emergency departments (EDs) but leave before being evaluated and treated is an important indicator of ED performance. This study examines patient- and hospital-level characteristics that may increase the risk of patients leaving EDs before being seen. The data are from the National Ambulatory Care Reporting System, an administrative database, and represent 4.3 million patient visits made to 163 Ontario EDs between April 2003 and March 2004. Among these data, the proportion that left without being seen (LWBS) was 3.1% (136,805). The rate of LWBS was highest among patients aged 15 to 35 years, those with less acute conditions and facilities that handle the highest volume of patients. Facility rates were positively correlated with facility median ED length of stay, annual facility volume and percentage of inpatient admissions. Understanding patient and facility characteristics that increase rates of LWBS may inform the process of developing measures to ensure timely access to ED care for all who seek it. PMID- 19305679 TI - The Manitoba First Nations Centre for Aboriginal Health Research: knowledge translation with Indigenous communities. AB - Knowledge translation at the Manitoba First Nations Centre for Aboriginal Health Research emerged through several population health research initiatives conducted in partnership with Canadian Indigenous peoples. An initial collaboration to carry out a regional health survey laid the foundation to share leadership, power and decision-making, and resulted in a program to build capacity in the First Nations health planning community through familiarity with the tools and skills of health researchers. Connections between health researchers and First Nations health planners have since led to the incorporation of evidence-based decision making in many First Nations communities. PMID- 19305680 TI - Organization and dissemination of working knowledge. AB - This commentary discusses the approach taken by the Manitoba Centre for Health Policy to systematize its working knowledge. Web-based knowledge tools facilitate maintenance of the information-rich environments present in almost every province. The malleability of administrative data means that variables can be defined in many different ways. Keeping track of what has been done becomes critical to facilitate reuse. New social data sets pose particular challenges and opportunities. Some more general issues of design are highlighted by a perusal of websites associated with other research efforts (the Panel Study of Income Dynamics, the National Bureau of Economic Research and the Institute for Clinical and Evaluative Sciences). PMID- 19305681 TI - The research collective: a tool for producing timely, context-linked research syntheses. AB - This paper reports on a research collective in primary healthcare (PHC) conducted in Quebec in 2004. A lead team of investigators synthesized 30 ongoing or recently completed studies from project description forms filled out by the participating researchers. The process of the collective is examined by addressing the three main challenges met in the course of its completion, namely, (a) the need to derive an analytical framework to regroup variables in a meaningful way, (b) the assessment of strength of evidence and (c) coping with a mix of quantitative and qualitative studies. Advantages of the collective over other forms of research synthesis include timeliness, low cost relative to the total cost of the studies it comprises and the information it generates and, finally, context linkage, which enhances relevance but which could limit transferability of the findings. Overall, the research collective appears to be a promising tool for research synthesis. PMID- 19305682 TI - The research collective: a model for developing timely, contextually relevant and dynamic approaches to research synthesis? AB - In this issue, Pineault and colleagues (2006) report on a Quebec-based research collective, a process aimed at rapidly synthesizing results from ongoing or recently completed research and releasing them to decision-makers. The process outlined differs markedly from Cochrane-based formal systematic reviews, and the authors claim a number of benefits that make the results more relevant to decision-makers, including enhanced timeliness of results, increased triangulation among emerging research findings, and more in-depth understanding of the impact of contextual environments on research results. This approach may offer opportunities to both advance and enrich existing synthesis tools. The research community should learn more from this process. PMID- 19305683 TI - The research collective: bridging the chasm. AB - Decision-makers in healthcare face many challenges in obtaining, analyzing and applying research results, including multiple priorities, competition among stakeholders and the time required to synthesize evidence. The research collective described by Pineault et al. appears to expedite access to information that can help inform policy decisions. It also challenges both researchers and decision-makers to better understand each other's context. Achieving such an understanding will require us all to step outside our traditional comfort zones, but this greater trust and flexibility stand to benefit all those who work in the healthcare system and those who rely upon it for care. PMID- 19305684 TI - Performance measurement in healthcare: part I--concepts and trends from a State of the Science Review. AB - OBJECTIVE: Performance measurement is touted as an important mechanism for organizational accountability in industrialized countries. This paper describes a systematic review of business and health performance measurement literature to inform a research agenda on healthcare performance measurement. METHODS: A search of the peer-reviewed business and healthcare literature for articles about organizational performance measurement yielded 1,307 abstracts. Multi-rater relevancy ratings, citation checks, expert nominations and quality ratings resulted in 664 articles for review. Key themes were extracted from the papers, followed by multi-reader validation. Information was supplemented with grey literature. RESULTS: The performance literature was diverse and fragmented, and relevant evidence was difficult to locate. Most literature is non-empirical and originates from the United States and the United Kingdom. No agreement on definitions or concepts is evident within or across disciplines. Study quality is not high in either field. Performance measurement arose in public services and business at about the same time. The evolution of thought on performance measurement ranges from unfettered enthusiasm to sober reassessment. CONCLUSIONS: The research base on performance measurement is in its infancy, and evidence to guide practice is sparse. A coherent multidisciplinary research agenda on the topic is needed. PMID- 19305685 TI - The impact of implementing managed competition on home care workers' turnover decisions. AB - This paper addresses the question: Did the implementation of managed competition in Ontario increase turnover in home care agencies? This question is addressed through a case study analysis of the impacts of tendering on the exiting home care labour force from three non-profit home care agencies during the period 1997 to 2001 in a mid-sized city in Ontario. These agencies provided 85% of the market share in 1996. Findings showed that 52% of the nurses and personal support workers (PSWs) left their agency over the five-year period. Analysis of the turnover data showed a temporal association between the implementation of managed competition and turnover. Additional support for the argument that the implementation of managed competition increased turnover is provided through analysis of a questionnaire sent to nurses and personal support workers who had left their agency during this period. Respondents indicated dissatisfaction with their pay, hours of work, benefits, heavy workload and lack of support from their supervisors/managers (all factors affected by the marketization of the home care sector) as reasons for leaving. Of those employed, only one-quarter remained in home care; most of those remaining were working in other healthcare fields such as hospitals and long-term care institutions. However, about one-third of employed PSWs were no longer working in the healthcare field. PMID- 19305686 TI - Consuming healthcare: what's in a word? PMID- 19305687 TI - From world war to class war: the rebound of the rich. AB - Incomes in Canada, as in many other countries, are becoming increasingly unequal. In North America this process has several notable features. First, after 40 years of stability, income has since 1980 been increasingly concentrated in the hands of the top 0.01% of earners. Second, this concentration correlates with an explosion in the relative earnings of corporate CEOs, a sort of "corporate kleptocracy." Third, the top earners have appropriated most of the productivity gains over this period. The resources and political influence of the super-rich underlie the growing prominence of the "elite" agenda: lower taxes, smaller government and privatization or shrinkage of social programs. The marketing of this agenda may explain much of the nonsense that contaminates health policy debates. PMID- 19305688 TI - Under the radar: stealth development of two-tier healthcare in Canada. AB - The shocked reaction of commentators to the recent Canadian Supreme Court decision (Chaoulli v. Quebec) overturning Quebec's ban on private healthcare insurance is difficult to square with the facts and policy options realistically open to provincial governments. The problem is that rhetoric has centred on preserving a single-tier universal system that has never existed in the form its supporters imagine. Meanwhile, quasi-private agencies and healthcare entrepreneurs have been improvising private care options, either ignored or abetted by governments. Consequently, policy and practice have become increasingly divergent. Supporters of Canadian-style medicare can only hope that the Chaoulli decision will force clearer-headed policy re-appraisal. Towards that end, this paper argues that provincial governments ought to focus more on robust regulation of already existing, privately financed healthcare, including the commissioning of care by Workers' Compensation Boards. PMID- 19305689 TI - Waits for surgery following hip fracture. AB - Almost all hip fracture patients undergo surgery to repair the fracture. Recent research suggests that timely repair is important for good outcomes following surgery. Patients who had surgical repair of a hip fracture in 2003-2004 were identified using hospitalization data collected by the Canadian Institute for Health Information. Time to surgery was calculated from day of admission to day of surgery. The associations of both patient and system characteristics with waits for surgery were considered. While the majority of patients had surgery on the day of or the day following admission, 29% waited two days or longer for surgery. Wait times were related to patients' age, hospital size, day of admission and whether patients were transferred. PMID- 19305690 TI - On the rock, in a hard place: challenges in working with advocacy and care provider groups. AB - Researchers from Memorial University of Newfoundland collaborated with a patient advocacy organization and the provincial cancer care provider on a study showing substantial variation in out-of-pocket costs for patients travelling to access cancer treatment. While the partnered research phase of the study proceeded smoothly, the successful dissemination of the findings--and the resulting government and media interest--created pressure for the provincial cancer care provider. This agency distanced itself from the study and ended its role in what has proved to be an otherwise successful partnership. PMID- 19305691 TI - Disseminating and organizing. AB - How do we reach a wide variety of possible readers? This commentary notes various ways in which academic information has been organized on the Web, stressing the usefulness of different formats and approaches. The Social Science Research Network provides a way for publications to become widely known at an early stage. Such Web-based facilities for one-stop searching are needed for academics and policy makers interested in health services, health policy, public health and population health. PMID- 19305693 TI - [Developing public health--primary care interfaces in Quebec: a case study]. AB - Public health agencies (PH) have various functions. One of them is to develop initiatives that will help decrease health inequalities among different groups in the population. To reach vulnerable groups, PHs must establish interfaces with various players, including primary care clinicians (PCC). Through a qualitative analysis, this case study identifies the organizational factors that either help or hinder the establishment of a functional interface in an area of the Quebec City region. Findings indicate that PHs must first undertake specific strategies to implement a new paradigm (such as the population perspective) among clinicians before introducing structural changes for developing and formalizing PH-PCC interfaces. PMID- 19305692 TI - Performance measurement in healthcare: part II--state of the science findings by stage of the performance measurement process. AB - OBJECTIVE: This paper summarizes findings of a comprehensive, systematic review of the peer-reviewed and grey literature on performance measurement according to each stage of the performance measurement process--conceptualization, selection and development, data collection, and reporting and use. It also outlines implications for practice. METHODS: Six hundred sixty-four articles about organizational performance measurement from the health and business literature were reviewed after systematic searches of the literature, multi-rater relevancy ratings, citation checks and expert author nominations. Key themes were extracted and summarized from the most highly rated papers for each performance measurement stage. RESULTS: Despite a virtually universal consensus on the potential benefits of performance measurement, little evidence currently exists to guide practice in healthcare. Issues in conceptualizing systems include strategic alignment and scope. There are debates on the criteria for selecting measures and on the types and quality of measures. Implementation of data collection and analysis systems is complex and costly, and challenges persist in reporting results, preventing unintended effects and putting findings for improvement into action. CONCLUSION: There is a need for further development and refinement of performance measures and measurement systems, with a particular focus on strategies to ensure that performance measurement leads to healthcare improvement. PMID- 19305694 TI - Priority setting in the Provincial Health Services Authority: case study for the 2005/06 planning cycle. AB - INTRODUCTION: Building on a survey of key decision-makers within the Provincial Health Services Authority (PHSA) of British Columbia, a process for prioritizing new service options within this organization for the 2005/06 budget cycle was developed and implemented by senior managers and researchers at the University of British Columbia. METHODS: A case study approach was taken in which development and implementation of the prioritization process was documented and feedback was obtained from decision-makers to evaluate the activity. Information from the literature was also used to identify areas for improvement. RESULTS: The 13 member Executive Committee of the PHSA conducted the prioritization. Criteria were defined and weighted, and options for new funding were explicitly rated against them. Multi-attribute decision software was used to rank options based on an overall benefit score. Cost-benefit ratios were also derived and program options were further ranked according to decision-makers' "gut-feel." Ranking between the methods was comparable, and each method contributed to the final decisions by the Executive Committee regarding which programs would receive limited new funding. CONCLUSION: Through a collaborative effort between decision makers and researchers, the PHSA was able to shift from allocating new resources on the basis of politics and bargaining to an evidence-driven, transparent, defensible process. Lessons learned from the current activity will help inform future priority setting in the PHSA and should provide insight for decision makers elsewhere. PMID- 19305695 TI - Health status and healthcare use patterns of rural, northern and urban Manitobans: is Romanow right? AB - OBJECTIVE: To compare health status and healthcare services use of rural, northern and urban Manitobans. METHOD: Using anonymized administrative claims data derived from the Population Health Research Data Repository housed at the Manitoba Centre for Health Policy, four Manitoba regions were compared--Winnipeg, Brandon, Rural South and North--for 1996/97-2000/01. Indicators include mortality and morbidity, ambulatory physician visit and specialist consultation rates, prevention/screening rates, selected surgery rates (cardiac catheterization, coronary artery bypass graft surgery, hip replacement) and "discretionary" surgery rates (tonsillectomy/adenoidectomy, Caesarean section, hysterectomy). Rates were annualized, directly standardized to the 1996 provincial population, and statistically tested for differences among the regions using Hotelling's T(2) statistic. RESULTS: Mortality and morbidity are high in the North, but the Rural South is average (except for high rates of injury mortality and stroke). Rural South and North have low ambulatory physician visits and specialist consultation rates, but high hospitalization rates compared to Brandon and Winnipeg. In prevention/screening rates, Rural South is variable and the North is low. For surgery rates, Rural South is variable, North is average, Brandon has below average surgical rates but high rates of discretionary procedures, and Winnipeg has high surgical rates but low discretionary procedures. Thus, "urban" is not necessarily synonymous with good health and better access to services, nor is "rural" or "remote" synonymous with poor health and inadequate healthcare. PMID- 19305696 TI - Income-based drug coverage in British Columbia: the impact on private and public expenditures. AB - BACKGROUND AND OBJECTIVES: In May 2003, the government of British Columbia adopted income-based pharmacare, replacing an age-based program. Stated policy goals included the reduction and reallocation of government spending. It was also hoped that income-based deductibles would increase consumer price sensitivity in decision-making. This analysis measured policy impacts on private and public expenditure and on expenditure drivers. METHODS: We employed a longitudinal research design using PharmaNet records of every prescription dispensed in the province from January 1996 to December 2004. Expenditure dynamics were analyzed using non-stochastic decompositions of trends. Analyses were stratified by five age categories and five socio-economic quintiles. The effect of the policy on expenditure trends and their sources was assessed using time series analysis. Additional analyses, using equivalent methods, were conducted using market-level data to compare per capita expenditure in British Columbia to the Canadian average over the period 1998-2004. RESULTS: The BC Ministry of Health was successful in reducing the public share of drug expenditure through the introduction of seniors' co-payments in 2002 and then income-based pharmacare in 2003. The policy change did not have major effects on aggregate expenditure trends in the province. While several statistically significant changes in expenditure dynamics occurred during the period of study, only an increase in seasonal "stockpiling" of medicines by seniors can reasonably be attributed to the policy changes. DISCUSSION: The lack of large and differential policy impacts on drug expenditure and utilization rates across age and income groups suggests that changes in the BC PharmaCare Program were designed in a manner that ensured continued access to medicines for the populations previously served by the drug plan (e.g., senior citizens). It also indicates that the policy did not significantly increase access to medicines by populations that might have been better served under the new policy (e.g., non-seniors). Finally, although it was hoped that income-based pharmacare might increase consumer cost consciousness, changes in the relative cost of certain drugs purchased following the policy change appear to have stemmed from other policies directly targeting the expenditure impact of therapeutic choices. PMID- 19305697 TI - Income-based drug coverage in British Columbia: the impact on access to medicines. AB - BACKGROUND AND OBJECTIVES: In May 2003, the government of British Columbia adopted income-based pharmacare, replacing an age-based program. Stated policy goals included the maintenance or enhancement of access to necessary medicines. This study examines the policy impact on access to two widely used drugs for chronic risk factors (antihypertensives and statins). METHODS: Data on incident antihypertensive and statin prescriptions between 1997 and 2004 were extracted from PharmaNet. Incident antihypertensive users were those who filled a first prescription after residing in the province for at least two years prior to the initial prescription date. The number of patients who ceased to fill a contiguous series of prescriptions (within 120 days of one another) was used as a measure of apparent discontinuation or interruption of therapy. We used time series analysis to test for changes in incident use and discontinuation. RESULTS: Between 1997 and 2004, 530,167 BC residents initiated therapy with an antihypertensive, and 264,904 BC residents initiated therapy with a statin. The 2003 policy change had no statistically significant impact on incident use of antihypertensives or statins, when stratified by age or income. Similarly, the 2003 policy did not change the rate of apparent discontinuations of therapy across age and income groups. However, a co-payment introduced in 2002 did increase end-of-year seasonality in apparent discontinuations in seniors--a finding that deserves further research. DISCUSSION: The 2003 transition to income-based pharmacare in British Columbia did not result in significant changes in access to, or continuation of, prescriptions to treat two leading chronic risk factors. PMID- 19305698 TI - Income-based drug coverage in British Columbia: the impact on the distribution of financial burden. AB - BACKGROUND AND OBJECTIVES: In May 2003, the government of British Columbia adopted an income-based pharmacare program, replacing the previous age-based program. Stated policy goals included improving the distribution of pharmaceutical payments across incomes. This analysis assesses the policy's effect on the distribution across incomes of both private payments and public subsidy for prescription drugs. METHODS: This analysis focuses on how the 2003 policy change affected the extent to which higher-income households pay a larger share of private drug expenditures and/or receive a smaller share of available public subsidies. Demographic information and drug spending data were extracted from BC PharmaNet and the BC PharmaCare Program for the years 2001-2004. These data were then graphed to assess (using concentration curves) changes in the progressivity of private and public pharmaceutical financing. RESULTS: Overall, the move to Fair PharmaCare resulted in larger but slightly less regressive private payments and smaller but slightly more progressive public subsidies. Because total drug spending increased while the total subsidy available decreased, average private household spending as a proportion of household income increased across virtually all age and income levels. DISCUSSION: The PharmaCare Program redistributed public subsidies in a manner that was more progressive than previous programs; this reduced the regressivity of private pharmaceutical payments. However, total public subsidy decreased, and private spending increased by a commensurate amount. This makes the program's overall financial impact on BC households somewhat ambiguous. Income-based pharmacare could improve financial equity unambiguously if public shares of drug spending are expanded. PMID- 19305699 TI - British Columbia's PharmaCare Program: fair or foul? PMID- 19305700 TI - Fat zombies, Pleistocene tastes, autophilia and the "obesity epidemic". AB - Canadians are fat and getting fatter: so say surveys up to and including the series of papers last August in Health Reports. By actual measurement, nearly a quarter of us (adults) are obese. So what? Obesity is clearly hazardous to health, but reports that 60% of us are "obese or overweight" border on fear mongering. A body mass index (BMI) over 25 is not a death sentence, and obesity will not bankrupt the healthcare system. The trends, though, are worrying. So will we rebuild cities--and, especially, suburbs--to be more pedestrian-friendly, suppressing auto-induced urban sprawl? Will we take on the fast-food industry as we did tobacco? Obesity is not destiny; Vancouver's rate is only half the national average. Canada could do better. PMID- 19305701 TI - Special Access Denied: A Case Study of Health Canada's Special Access Program. AB - This paper examines the ethical probity of Health Canada's Special Access Program (SAP). The SAP is designed for patients with life-threatening conditions who require "emergency" access to drugs that are not authorized for use in Canada when conventional therapies have failed. We argue that the SAP inappropriately uses the tenets of evidence-based decision-making in situations where evidence based decision-making is unfeasible. The SAP should abandon its pretence of evidence-based decision-making and adopt a transparent process in which the values guiding decisions are explicit and corrigible. We recommend the ethical principles of autonomy, non-maleficence, beneficence and justice. PMID- 19305702 TI - Pandemic threats and the need for new emergency public health legislation in Canada. AB - The 2003 outbreak of Severe Acute Respiratory Syndrome (SARS) exposed serious limitations in Canada's ability to respond to a public health emergency. Considerable progress has been made since SARS in addressing these limitations, including the creation of the new Public Health Agency of Canada. A remaining contentious question is whether there is a need for new federal emergency public health powers. Approaches to public health problems are best handled through collaborative processes, recognizing the critical importance of the local public health response. Nevertheless, this paper argues that a legislative back-up plan must be available to the federal government in the event that collaborative relationships break down. At the minimum, legislation should give the federal government the authority to have guaranteed access to surveillance data during a public health emergency. The legislation should also consider providing the federal government with the authority to devote the nation's resources to the management of an emergency at its earliest stages. However, any legislative approach must be combined with the development of appropriate capacity at the national level to ensure that new powers can be adequately utilized and that required funding reaches public health officials at other levels of government. PMID- 19305703 TI - Waiting for care in Canada: findings from the health services access survey. AB - Waiting for care has been and continues to be a major issue for the healthcare sector in Canada. While considerable gains have been made regarding valid and reliable information on waiting times, gaps remain. Statistics Canada continues to provide information regarding patients' experiences in accessing care at the national and provincial levels, including how long individuals waited for specialized services, through the Health Services Access Survey. The survey offers several advantages, including waiting time information that is comparable across time and space, enhanced patient information and information regarding patients' experiences in waiting for care. The results for 2005 indicate that median waiting time for all specialized services was between 3 and 4 weeks and remained relatively stable between 2003 and 2005. Waiting times for specialist visits did not vary by income. In addition to being asked how long they waited, individuals were asked about their experiences in waiting for care. While the majority of individuals waiting for care indicated that their waiting time was acceptable, there continues to be a proportion of Canadians who feel they are waiting an unacceptably long time for care. Between 11% and 18% of individuals waiting for care indicated that their life was affected by waiting. PMID- 19305704 TI - Knowledge Translation through Research-Based Theatre. AB - This case documents the metamorphosis of an academic public health project on the implementation of Ontario's return-to-work policies and practices into a community theatre project, the production of a play called Easy Money. The result was a highly successful knowledge translation initiative. Injured workers, the focus of the academic study, intensely identified with the play and were given hope that their individual experiences could be broadly translated. For the researchers, the undertaking provided new perspectives on the original research problem, validated their original findings and generated numerous topics for subsequent research. PMID- 19305705 TI - A Cautionary Tale of Downloading Public Health in Ontario: What Does It Say about the Need for National Standards for More Than Doctors and Hospitals? AB - The 2003 SARS outbreak highlighted the importance of maintaining an adequate public health (PH) infrastructure, and cast doubt on the wisdom of basing the system locally without adequate provisions for higher-level oversight and coordination. Structurally, it highlighted the policy legacy of the 1998 Ontario decision to download full responsibility for funding PH services to municipal governments, forcing such services into budgetary competition with the "hard" services traditionally provided by local government. The federal role in PH has traditionally been minimal; PH was never included as a mandatory service in the Canada Health Act, while reform proposals have focused upon such admittedly important directions as pharmacare and home care rather than PH. Although PH has moved up the policy agenda, with a focus on pandemic preparedness, the Ontario events suggest a pressing need for setting national and provincial/territorial standards for PH, and developing mechanisms for enforcing them. PMID- 19305706 TI - Canadian community health survey: major depressive disorder and suicidality in adolescents. AB - BACKGROUND: Contrary to other developed countries where adolescent suicide rates have declined in the last decade, the rate in Canada has remained unchanged. Suicide is the second leading cause of death in Canadian adolescents and poses a serious public health concern. However, there is little epidemiological data examining the rates of suicidality or depression - two factors most closely associated with completed suicides. This study therefore examines the rates of depression and suicidality in adolescents aged 15-18. METHODS: Data from the Canadian Community Health Survey Cycle 1.2 on Mental Health and Well-being, a population-based survey conducted by Statistics Canada, were used to examine the rates of depression and suicidality in adolescents aged 15-18. Lifetime prevalence rates were calculated for depression and suicidality by region for males and females. Multivariate analyses were conducted to test the robustness of these results. RESULTS: The lifetime prevalence rates were 7.6% for depression and 13.5% for suicidality. There were significant gender differences for both: 4.3% of males and 11.1% of females had depression, and 8.8% of males and 18.4% of females had suicidality. After adjustment for age, sex and household income, the Maritimes had a lower rate of depression and British Columbia had a higher rate of suicidality relative to Ontario. Youth from low-income households had a higher risk of suicidality. INTERPRETATION: The findings suggest that depression and suicidality are common in adolescents and that females are more likely to be affected. The results also point to regional and socio-economic differences. Future research should examine differences that exist in mental health services provision and access. This will aid in the development of national, regional and local strategies to address the issue of depression and suicidality in Canadian adolescents. PMID- 19305707 TI - Income-based drug coverage in british columbia: towards an understanding of the policy. AB - BACKGROUND AND OBJECTIVES: In May 2003, the government of British Columbia adopted income-based pharmacare, replacing an age-based drug benefits program. Income-based pharmacare has been proposed as a national model for Canada. Prior to evaluating the policy impact of the change, we sought to understand classic elements of the policy process: the problem that prompted the policy change, the objectives to be met, the choice of policy instruments and potential measures of success or failure. METHODS: We conducted interviews with decision-makers in British Columbia shortly after the policy was introduced. We used purposive sampling to target individuals specifically involved with BC pharmacare policy, including current or former government employees, government ministers and the leader of the opposition party. Seventeen of 24 invited subjects participated. RESULTS: Participants identified primary and secondary problems that required remedy through policy change: financial pressures and equity issues. Financial pressures were framed in the context of budgetary constraints on the BC PharmaCare Program, with reduction in public spending cited as the primary policy objective. Participants also indicated that it was important to minimize potential harm from any program change and, ideally, to improve access for low income families. DISCUSSION: Income-based pharmacare in British Columbia appears to be the result of a two-stage policy-making process. Budgets were set by pan ministerial actions of the provincial government. In turn, these budgetary measures constrained policy design at the ministerial level. Income-based coverage was BC PharmaCare's choice among options that would meet budget expectations. Success or failure of the policy would be gauged by (a) meeting budget targets, (b) maintaining or increasing access to medicines and (c) improving financial equity. PMID- 19305708 TI - Income-Based Drug Coverage in British Columbia: Lessons for BC and the Rest of Canada. AB - BACKGROUND: In May 2003, the government of British Columbia adopted income-based pharmacare, replacing an age-based drug benefits program. Stated policy goals included reducing government spending, maintaining or enhancing access to medicines and improving financial equity. The province's experience on these policy dimensions can inform policy making in other jurisdictions and offers insight into priorities for Canada's National Pharmaceuticals Strategy. METHOD: The research team created an anonymized database with information about drug use, private and public expenditure and household income for all residents of British Columbia from 1996 to 2004. This database was used to evaluate the impact of the policy on trends in drug expenditures, utilization and sources of payment for seniors and non-seniors of different income levels. RESULTS: In the immediate term, Fair PharmaCare appears to have met many of its policy goals. Government spending was reduced. Access to medicines was maintained (though not enhanced). And the distributions of private and public expenditures were brought more closely in line with distribution of income. Long-run impacts depend largely on how a reduced role for government affects trends in costs, access and equity. Early indications suggest that a larger role for government may be needed to maintain performance on desired policy objectives over time. CONCLUSION: In the long run, there is reason for setting a new national standard for pharmacare that increases, not decreases, the share of publicly covered spending in every province. The federal government could play a key role by helping provinces increase public funding for prescription drugs and thereby facilitate cost control, maintain access to medicines and enhance financial equity. PMID- 19305709 TI - Healthcare use of families of injured workers before and after a workplace injury in british columbia, Canada. AB - OBJECTIVES: To examine the overall healthcare and mental healthcare services use of families of injured workers before and after a workplace injury. METHODS: We use an administrative database that links individual publicly funded healthcare data and Workers' Compensation Board (WCB) data for the entire population of British Columbia (BC), Canada. The spouses and children of all injured workers who filed a WCB claim in 1994 and missed one or more days of work due to the injury (lost time) were included. We compare their change in use of healthcare services relative to a year before the injury to families of workers who did not require time off for their injuries (no lost time) and families of individuals who were not injured (non-injured comparisons). RESULTS: Differences in healthcare services use among the three groups of spouses were marginal, and differences for increases in mental healthcare services use were non-significant. As well, all three groups of children decreased their use of physician and hospital services and increased their use of mental healthcare services, with very little difference among groups. CONCLUSION: This was a descriptive study looking at a broad group of injured workers and their families. Even modest increases in healthcare use following a workplace injury have some basis for further study. PMID- 19305710 TI - Determinants of unacceptable waiting times for specialized services in Canada. AB - BACKGROUND: Much of the current evidence regarding timely access to healthcare services focuses on the duration of the waiting time as the principal determinant of wait time acceptability. We conducted the first national-level analysis of wait time acceptability in Canada to identify the determinants of unacceptable waits for specialized healthcare services, including selected demographic and socio-economic variables. METHODS: We analyzed data reported by respondents to a national survey on access to healthcare services who accessed specialized services (i.e., specialist visits, non-emergency surgery and selected diagnostic tests) during a 12-month period. We used univariate analyses and weighted logistic regression to examine the relation between wait time acceptability and selected demographic, socio-economic and health status factors for each specialized service. RESULTS: Between 17% and 29% of patients who waited for a specialized service declared that their waiting time was unacceptable. Most individuals reported waiting less than 3 months for their services. Between 10% and 19% of those who waited indicated that waiting for care affected their lives. Results of the logistic regression analyses showed that longer waits and adverse experiences during the waiting period were significantly associated with higher odds of reporting an unacceptable waiting time for all three types of specialized services. The role of socio-economic and demographic factors on wait time acceptability was varied. Individuals with lower education were consistently less likely to consider their waiting times unacceptable. Patients less than 65 years of age were more likely to consider their waiting times unacceptable for specialist visits and diagnostic tests. INTERPRETATION: Our study shows that the primary determinants of waiting time acceptability are the length of the waiting time and the effects of waiting on the patient's life. In addition, some patient characteristics, such as age and education, may play a role, pointing to the potential role of patient expectations in determining the acceptability of waits for specialized services. PMID- 19305711 TI - Determinants of waiting time for a routine family physician consultation in southwestern ontario. AB - Waiting times are a reality in Canada's publicly financed single-payer healthcare system. While there are ample data about waiting times for specialized investigations and procedures, few data exist about waiting times to see family physicians, and determinants of this wait. We analyzed data from a survey of 731 family physicians in southwestern Ontario to understand physician- and practice level determinants of waiting time. Physician gender, usual number of patients seen per week, involvement in teaching and population served were the key determinants of physician-reported waiting time. PMID- 19305712 TI - Wait times for paediatric rehabilitation. AB - BACKGROUND: Early therapy intervention for children with disabilities may improve functional outcomes. Access to paediatric rehabilitation services can sometimes be difficult. OBJECTIVES: To describe waiting time to receive physical therapy (PT) and occupational therapy (OT) services at rehabilitation centres for young children with physical disabilities; to examine factors associated with these waiting times. DESIGN: Prospective cohort. SUBJECTS: Two hundred and six children with physical disabilities, aged 6 to 72 months, referred in 2002-2004 from the Montreal Children's Hospital and Sainte-Justine Hospital to paediatric rehabilitation centres. MEASURES: Data on date of referral, age, gender and diagnosis were obtained from the hospital databases. Data on date of first PT or OT appointments at the rehabilitation centre, family socio-demographics and disability severity (WeeFIM) were obtained during parental interviews. RESULTS: Half of the sample waited more than 7 and 11 months for PT and OT services, respectively. Shorter waiting time was significantly associated with younger child's age and referral to one particular rehabilitation centre. CONCLUSION: Children with physical disabilities experience long waiting times for PT and OT rehabilitation services. Strategies to improve timely service delivery are needed. PMID- 19305713 TI - Cracks in the Foundation: The Precarious State of Canada's Primary Care Infrastructure. PMID- 19305714 TI - Green leviathan? Thomas hobbes, joel bakan and Arnold schwarzenegger. AB - Thomas Hobbes postulates that men are driven by "a perpetual and restless desire of power after power, that ceaseth only in death." The miserable consequences of this drive for power and the competing "desire of ease and sensual delight" and "fear of death and wounds" lead them to establish and obey. Substituting "profit" for "power" yields a description of the modern corporation, but without the desires or fears of natural persons. Such "unnatural persons" lack the Hobbesian ground of obligation, yet have appropriated the privileges and protections of natural persons. They challenge or undermine the sovereign wherever it limits their profits. Governor Schwarzenegger's re-election in California, however, on a strong anti-CO(2) program, suggests a willingness by threatened natural persons to re-empower Leviathan. PMID- 19305715 TI - Data protection and the promotion of health research. AB - This paper challenges the argument that data protection legislation may harm research by unduly restricting the flow of personal health information. I unpack the assumption that privacy is an individual right that must give way to research as a social good, and explore how data protection laws facilitate the flow of information for research purposes. I conclude that researchers should embrace data protection laws because they help construct trust in research practices, mitigate the commercial imperatives that flow from the fact that research is a public-private enterprise and protect the accuracy of data. Good research design should recognize that privacy is a social value and an essential element of psychological health and social relationships. And since research databases do not exist in isolation, researchers must respect the fact that the non-consensual flow of information poses risks of harm, including the secondary use of health research databases for social control, that must be managed. PMID- 19305716 TI - Data protection and the promotion of health research: if the laws are not the problem, then what is? AB - Data protection laws offer such broad exemptions for research that research ethics boards and data custodians lack sufficient guidance as to when it may be acceptable to release data to researchers without consent. The result: idiosyncratic institutional policies that create major challenges for researchers conducting multi-centred studies. The 2005 CIHR Best Practices for Protecting Privacy in Health Research provide an important first step towards greater clarity. However, there is still a need to translate these Best Practices into harmonized policies. This should be seen as an opportunity rather than a threat. Clear rules for data protection will reinforce public trust, which is essential for continued access to personal information for research. PMID- 19305717 TI - Data protection and the promotion of health research. PMID- 19305718 TI - Connectedness within Social Contexts: The Relation to Adolescent Health. AB - International studies have shown that the more developmental assets adolescents possess, the greater their likelihood of engaging in health-enhancing practices and the lesser their likelihood of engaging in practices that put health at risk. Logistic regressions were conducted on data from the 2000-2001 National Longitudinal Survey of Children and Youth (NLSCY) for 12- to 15-year-old Canadian youth to examine which of five assets accounts for the most variance in positive health outcomes and participation in risky health behaviours. Connectedness within social contexts, particularly to family and school, was associated with several self-reported positive health outcomes and behaviours (excellent or very good health, high self-worth, and less alcohol, tobacco and marijuana use). Connectedness among peers was associated with better self-assessed health and higher self-worth but also with more use of alcohol, tobacco and marijuana. A comprehensive approach to healthy youth development that emphasizes and increases positive relationships in these contexts may facilitate the transition of Canadian youth into healthy adulthood. PMID- 19305719 TI - Wait time benchmarks, research evidence and the knowledge translation process. AB - The first set of evidence-based benchmarks for medically acceptable wait times, announced in December 2005, were developed, in part, through a novel partnership between the Provincial and Territorial Ministries of Health, the Canadian Institutes of Health Research (CIHR) and Canada's health services research community. Responding to a direct request for assistance and demanding timelines from the Provincial and Territorial Ministries of Health, CIHR mounted a rapid response funding process and supported eight Canadian teams to synthesize evidence to inform the development of the first set of benchmarks. This experience demonstrated that both the research funding process and research syntheses themselves can rapidly inform policy making in even the most heated of environments. PMID- 19305720 TI - The effect of pharmaceutical patent term length on research and development and drug expenditures in Canada. AB - While pharmaceutical patent terms have increased in Canada, increases in patented drug spending have been mitigated by price controls and retrenchment of public prescription drug subsidy programs. We estimate the net effects of these offsetting policies on domestic pharmaceutical R&D expenditures and also provide an upper-bound estimate on the effects of these policies on Canadian pharmaceutical spending over the period 1988-2002. We estimate that R&D spending increased by $4.4 billion (1997 dollars). Drug spending increased by $3.9 billion at most and, quite likely, by much less. Cutbacks to public drug subsidies and the introduction of price controls likely mitigated drug spending growth. In cost benefit terms, we suspect that the patent extension policies have been beneficial to Canada. PMID- 19305721 TI - The effect of pharmaceutical patent term length on r&d and drug expenditures in Canada. AB - The authors make a useful and original contribution to evaluating the impact of strengthened patent protection of pharmaceuticals on Canadian healthcare expenditures. This commentary argues that their second task - measuring the impact of strengthened patent protection on R&D spending in Canada - faces an insurmountable conceptual problem: Corporate decisions on the location of R&D activities bear no theoretical relationship to the location of sources of R&D funding and are guided by other factors, including political considerations. PMID- 19305722 TI - Do patent terms impact domestic r&d spending in the pharmaceutical industry? AB - Grootendorst and Di Matteo's study showed that extended patent terms in Canada significantly increased domestic R&D spending in the pharmaceutical industry. However, some of the authors' assumptions and methods, including the choice of control variables, the technique used in calculating policy impacts over time and the failure to incorporate the influence of global factors, are problematic. The overall impression is that the study highlights a correlation between extended patent terms and increased R&D expenditure in Canada but does not provide firm evidence of a causal link. PMID- 19305723 TI - Response to pazderka and schroeder. PMID- 19305724 TI - Governance, health policy implementation and the added value of regionalization. AB - OBJECTIVES: In this paper we focus on governance and the added value of regionalization in the context of health policy implementation. What are regional boards' patterns of action in the governance process?How do these patterns favour policy implementation? ANALYTICAL FRAMEWORK: To enhance our understanding of the role of regional boards in governance processes, we relied on four conceptual constructs that corresponded to models of collective action: political, technocratic, democratic and cognitive. Alongside the four models, we analyzed the impact of governance on health policy implementation using Mazmanian and Sabatier's general analytical framework, which identifies three types of variables that affect public policy implementation: (1) variables related to the complexity of the problem, (2) statutory variables that structure the implementation of the policy and (3) non-statutory variables related to the context. METHODS: We conducted a qualitative, longitudinal case study of the regional implemention of the Program to Combat Cancer in Quebec. FINDINGS: This research stresses the added value of a clinico-administrative governance of change, whereby regional boards, in synergy with clinical leaders, participate in the orientation of collective action. Analysis of the regional board's patterns of action reveals the utility of combined technocratic, democratic, political and cognitive actions. PMID- 19305725 TI - Advancing Health Promotion Priorities: Stories of Capacity Building from the Canadian Heart Health Initiative (CHHI). AB - The purpose of this paper is to explore how public health professionals built capacity to carry out health promotion despite a low level of investment and competition for financial resources with acute-oriented healthcare services. Three data sources are used in this analysis: key-informant interviews with project participants, final reports from three provincial Heart Health projects in Canada (Prince Edward Island, Ontario and Manitoba) and major provincial health policy documents prior to and during each project. We use a narrative policy analysis to identify contextual factors influencing health promotion priority and progress through capacity building. Common capacity building themes emerged from the data despite the different contexts within which the projects were situated: building community trust and support, developing a linking system that promotes provincial partnerships and assisting in sustainability efforts by coordinating resources and efforts towards a common chronic disease prevention strategy. Each of these provincial projects overcame instances of resistance to advancing a health promotion agenda by concentrating on building relationships, by making better use of existing structures and organizations and by developing new productive unions that shared a primary prevention agenda. PMID- 19305726 TI - Innovations in the Ethical Review of Health-Related Quality Improvement and Research: The Alberta Research Ethics Community Consensus Initiative (ARECCI). AB - The Alberta Research Ethics Community Consensus Initiative (ARECCI) is a unique Canadian initiative that addresses the ethical oversight of two main categories of health-related investigative projects: research and quality improvement (including quality assurance and program evaluation). ARECCI was formed as a result of discussions arising from health regions, health researchers and the Alberta Committee of Research Ethics Boards (REBs) Chairs, who all desired a clearer and more consistent approach to the ethical oversight of investigative health projects. The Alberta Heritage Foundation for Medical Research (AHFMR) established and supported ARECCI in 2003 in response to this need. ARECCI is unique in its ongoing efforts to bring together a wide-ranging group of stakeholders to develop consensus on a set of pragmatic recommendations and tools for the ethical review of research and quality improvement, and to get extensive consultation on those recommendations. This paper presents the ARECCI context and process, recommendations and tools produced by ARECCI and lessons learned from the ongoing ARECCI process. PMID- 19305727 TI - To Count Heads or to Count Services? Comparing Population-to-Physician Methods with Utilization-Based Methods for Physician Workforce Planning: A Case Study in a Remote Rural Administrative Region of British Columbia. AB - OBJECTIVES: To demonstrate the feasibility of a population-based measure of physician services utilization by type of service as a tool for physician workforce planning. SETTING: The Northern Health Region of British Columbia. DESIGN: Retrospective descriptive statistics are compiled about the regional population's physician services utilization by the specialty of the service, irrespective of the specialty or location of the provider. These are compared to norms based on provincial average utilization, adjusted for age and sex, and norms based on population-to-physician ratio recommendations. METRICS: By specialty type of service: actual utilization; age-sex expected utilization; in region, out-of-region and out-of-province utilization; full-time equivalency (FTE) values of actual and expected utilization; and FTE requirement to meet a set of recommended population-to-physician targets. Specialty substitution by general practitioners (GPs) is also quantified. RESULTS: The overall estimated deficit in physician numbers is similar between the two methods (51 versus 54), but the magnitude of surplus or deficit by specialty is greater with the population-to-physician method. CONCLUSION: The method targets an equitable distribution, rather than normative ideal physician supply. The magnitude of estimated surplus or deficit at the level of each specialty is greater with the population-to-physician ratio approach. The latter fails to consider interregional flow and specialty substitution. A population-based utilization approach is demonstrated to be a feasible, and in many ways superior, tool for physician resource planning. PMID- 19305728 TI - Involving decision-makers in producing research syntheses: the case of the research collective on primary healthcare in quebec. AB - This paper reports on a research collective on primary healthcare (PHC) conducted in Quebec in 2004. Thirty ongoing or recently completed studies were synthesized through a process involving a high degree of exchange among researchers who conducted the original studies, investigators and decision-makers. The viewpoints expressed by decision-makers who participated in the process were analyzed in terms of convergence with and divergence from the researchers' viewpoints. In four cases, there was convergence between the decision-makers' and the researchers' viewpoints, thus increasing the validity of the collective's findings. The main divergence between the two groups' viewpoints concerns the strategy adopted in Quebec to create local health and social services networks. Such divergence reflects the distinction made by Klein between scientific evidence and organizational and political evidence.Our study results illustrate that decision-makers' viewpoints can play an important interpretive and complementary role in producing research syntheses. Although integrating decision makers' viewpoints into syntheses has been regarded as a strategy for improving the use of research findings, our analysis shows that decision-makers' view points do not necessarily have to be integrated into syntheses but can, instead, be examined for convergence with or divergence from researchers' viewpoints. This deliberative process can enrich discussions and lead to enlightened decision- and policy making. PMID- 19305729 TI - Divvying up the funding pie: what share for health services and policy research? PMID- 19305730 TI - Mr. Harrington, self-rated health and the canadian chicken. AB - The simplest way to find out how healthy people are is to ask them. The question: "Would you rate your health as excellent, very good, good, fair or poor?" is quick and easy (and cheap) to administer, and correlates well with more objective indicators of health (such as subsequent death). But there are significant cross national differences in response patterns. Canadians are much less likely than Americans to provide extreme responses - excellent or poor. International comparisons yield more striking examples. Americans rate their health among the highest in the OECD, despite mortality measures that are among the worst. The Japanese, with the world's best mortality measures, rate their health near the bottom. Can self-reports be standardized for these cultural effects? PMID- 19305731 TI - The new general practice contract and reform of primary care in the United kingdom. AB - In April 2004, the United Kingdom introduced a new General Medical Services (GMS) contract that provided new governance and incentive arrangements for general practice. In particular, the new Quality and Outcomes Framework is a points-based system that sets targets for clinical, organizational and practice-related standards with financial payments for achieving set levels of performance. This paper describes the new contract arrangements and their impact on general practice - focusing on the experience in England, where wider policy changes are also having an important impact on practices - and drawing out potential lessons that will be of interest to Canadian practitioners and policy makers. PMID- 19305732 TI - Shopping for high-technology treatment in another province. AB - In order to address long healthcare waits, political and professional groups have recommended sending patients to other provinces for diagnostic procedures or treatment. We investigated the feasibility of such recommendations, specifically, whether residence in one province can impede access to MRIs in another province. We contacted all public MRI facilities in Canada and found no difference in wait times between prospective in- and out-of-province patients, although wait times were highly variable from province to province. Over one-fifth (19/86=22%) of centres imposed barriers for out-of-province patients to access care. We discuss several jurisdictional, financial and logistic considerations regarding the feasibility and appropriateness of implementing a national strategy of interprovincial patient transfer for healthcare. PMID- 19305733 TI - Home care evolution in alberta: how have palliative clients fared? AB - This study compared palliative and non-palliative home care clients, services and providers, and described changes over a decade of health system reform (1991/92 2000/01). Complete individual-anonymous data from Alberta's home care database were analyzed. Over these 10 years, 7.0% of all home care clients were classified as palliative. The proportion of home care clients who were classified as palliative varied from 2.2% to 9.6% among health regions. The number of palliative clients more than doubled, although this growth was less than that of short-term clients. Home support aides were the most common home care provider, and personal care was the most common service provided to all clients. Although the average number of care hours prior to death for palliative clients increased from 40.9 to 87.9 hours, the relatively small amount of home care provided to dying persons raises concerns about informal caregiver burden and possible overreliance on hospitals to provide end-of-life care. PMID- 19305734 TI - A community-researcher alliance to improve chronic wound care. AB - A partnership between health services researchers from Queen's University and the University of Ottawa, a community nursing agency and a home care authority in Ottawa led to major improvements in the quality of care for people with leg ulcers. The synthesis of both external and local evidence played a key role in the adoption of an evidence-based protocol and provided the critical context to support a significant reorganization of the existing service delivery model. This case demonstrates that, with a collaborative partnership approach, systematic and transparent research processes can be rapidly developed to support policy change. PMID- 19305735 TI - Why equity in financing first nations on-reserve health services matters: findings from the 2005 national evaluation of the health transfer policy. AB - BACKGROUND: This paper reports on selected findings from the 2005 National Evaluation of the Health Transfer Policy. Three hypotheses were tested, namely: (1) that inequalities in per capita financing exist between First Nations organizations, (2) that variations in per capita funding among communities cannot be explained by variations in the program responsibilities each assumed and (3) that First Nations organizations that transferred in the early 1990s now have access to fewer resources on a per capita basis than those that transferred more recently. METHODS: We compared (1) the per capita funding for 30 medium-sized communities (population = 401-3,000) that have Health Centres and the 13 similarly sized communities that have Health Stations, (2) program responsibilities and per capita funding for the same 30 communities and (3) the relationship between 2001-2002 per capita funding and the year of transfer for the same communities. We used data provided to us by the First Nations and Inuit Health Branch of Health Canada from 1989 to 2002. RESULTS: The results show that differences in per capita funding exist among and within regions. These differences cannot be explained by the responsibilities each community chose to assume. Differences are also related to the year First Nations entered into a transfer agreement. CONCLUSIONS: We recommend that formula-based financing be adopted to reduce inequalities. Such a formula should reflect needs, population growth and changes in costs of service delivery. PMID- 19305736 TI - The Relationship between Characteristics of Home Care Nursing Service Contracts under Managed Competition and Continuity of Care and Client Outcomes: Evidence from Ontario. AB - The purpose of this study was to investigate the impact of the Request for Proposal (RFP) process - specifically, the profit status of provider agencies awarded contracts, the service volume awarded and contract duration - on the quality of home care services and outcomes. A cross-sectional (contract characteristics) and repeated measures (clients) design was used to collect data on the study variables. Primary data were collected in 2002-2003 from 11 Community Care Access Centres (CCACs) and 11 nursing provider agencies in Ontario. The sample included 750 home care clients recruited consecutively from home care referrals. Follow-up assessments were completed on 498 clients. CCACs and provider agencies completed written questionnaires about profit status, contract volume, duration of contract, potential for renewal, number of visits made by a principal nurse and number of visits made by a registered nurse. Data were collected on client health outcomes either at admission to home care service for new clients or at entry to the study for long-term clients, and then at discharge from service or at the end of six weeks, whichever came first, using the eight subscales of the Medical Outcome Study SF-36. Analysis was conducted using hierarchical linear modelling.For the most part, contract characteristics were not related to the consistency of principal nurse visits or client outcomes. Where differences existed, they were small. Clients of agencies awarded longer contracts received greater consistency in principal nurse visits than clients of agencies awarded shorter contracts. Clients cared for by for-profit agencies reported slightly higher satisfaction with care and better mental health outcomes than clients cared for by not-for-profit agencies. The percentage of visits made by a registered nurse was positively associated with social function outcome for clients at follow-up. In conclusion, the study findings suggest that contract characteristic variables had a small effect on home care client outcomes. PMID- 19305737 TI - Notice of compliance with conditions: a policy in limbo. AB - Since 1998, the Therapeutic Products Directorate (TPD) has had a policy termed the Notice of Compliance with conditions (NOC/c) in order to allow earlier marketing of promising drugs for serious conditions before the drugs have definitively demonstrated clinical efficacy. Drugs approved under the NOC/c must undergo post-marketing trials to show clinical benefits. The reasons that some drugs receive a NOC/c are not always apparent, and the TPD releases only general information regarding the conditions that need to be fulfilled. Some drugs have fulfilled their conditions in under 1.4 years, but others had unfulfilled conditions after seven years. Doctors may not be aware that drugs are marketed with a NOC/c nor that some drugs have had their NOC/c withdrawn, and as a consequence may be prescribing inappropriately for their patients. Other jurisdictions have similar policies but with stricter and more transparent requirements. Adopting these provisions, along with other reforms, could help ensure that the NOC/c policy meets it objectives. PMID- 19305738 TI - Medicare financing and redistribution in british columbia, 1992 and 2002. AB - Equity in healthcare in British Columbia is defined as the provision of services based on need rather than ability to pay and a separation of contributions to financing from the use of services. Physician and hospital services in Canada are financed mainly through general tax revenues, and there is a perception that this financing is progressive. This paper uses Gini coefficients, concentration indexes and Kakwani indexes of progressivity to assess the progressivity of medicare financing in British Columbia in 1992 and 2002. It also measures the overall redistributive effect of medicare services, considering both contributions to financing and use of hospital and physician services. The conclusion is that medicare does redistribute across income groups, but this redistribution is the result solely of the positive correlation between health status and income; financing is nearly proportionate across income groups, but use is higher among lower-income groups. Informed public debate requires a better understanding of these concepts of equity. PMID- 19305739 TI - Human 5-HT(4) and 5-HT(7) receptor splice variants: are they important? AB - G-protein-coupled receptors (GPCRs), which are encoded by >300 genes in the human genome, are by far the largest class of targets for modern drugs. These macromolecules display inherent adaptability of function, which is partly due to the production of different forms of the receptor protein. These are commonly called 'isoforms' or 'splice variants' denoting the molecular process of their production/assembly. Not all GPCRs are expressed as splice variants, but certain subclasses of 5-HT receptors are for example, the 5-HT(4) and 5-HT(7) receptors. There are at least 11 human 5-HT(4) and three h5-HT(7) receptor splice variants. This review describestheir discoveries, nomenclature and structures. The discovery that particular splice variants are tissue specific (or prominent) has highlighted their potential as future drug targets. In particular, this review examines the functional relevance of different 5-HT(4) and 5-HT(7) receptor splice variants. Examples are given to illustrate that splice variants have differential modulatory influences on signalling processes. Differences in agonist potency and efficacies and also differences in desensitisation rates to 5 HT occur with both 5-HT(4) and 5-HT(7) receptor splice variants. The known and candidate signalling systems that allow for splice variant specific responses include GPCR interacting proteins (GIPs) and GPCR receptor kinases (GRKs) which are examined.Finally, the relevance of 5-HT receptor splice variants to clinical medicine and to the pharmaceutical industry is discussed. PMID- 19305740 TI - Anti-inflammatory and immune therapy for Alzheimer's disease: current status and future directions. AB - From the initial characterizations of inflammatory responses in Alzheimer's disease (AD) affected brains, namely the demonstration of activated microglia and reactive astrocytes, complement system activation, increased production of proinflammatory cytokines, and evidence for microglial-produced neurotoxins, there was hope that reducing inflammation might be a feasible treatment for this memory-robbing disease. This hope was supported by a number of epidemiology studies demonstrating that patients who took non-steroidal anti-inflammatory drugs had significantly lower risk of developing AD. However, clinical trials of anti-inflammatories have not shown effectiveness, and in recent years, the concept of immune therapy has become a treatment option as animal studies and clinical trials with Abeta vaccines have demonstrated enhanced amyloid removal through stimulation of microglial phagocytosis.This review will examine the current status of whether inhibiting inflammation is a valid therapeutic target for treating AD; what lessons have come from the clinical trials; what new pathways and classes of agents are being considered; and how this field of research can progress towards new therapeutics. We will examine a number of agents that have shown effectiveness in reducing inflammation amongst other demonstrated mechanisms of action. The major focus of much AD drug discovery has been in identifying agents that have anti-amyloid properties; however, a number of these agents were first identified for their anti-inflammatory properties. As drug development and clinical testing is a costly and lengthy endeavor, sound justification of new therapeutic targets is required. Possible future directions for AD anti-inflammatory or immune clearance therapy will be discussed based on recent experimental data. PMID- 19305741 TI - The ERK 1 and 2 pathway in the nervous system: from basic aspects to possible clinical applications in pain and visceral dysfunction. AB - The extracellular signal-regulated kinases 1 and 2 (ERK) cascade, member of the mitogen-activated protein kinases superfamily of signalling pathways, is one of the best characterized pathways as many protein interactions and phosphorylation events have been systematically studied. Traditionally, ERK are associated with the regulation of proliferation and differentiation as well as survival of various cell types. Their activity is controlled by phosphorylation on specific aminoacidic residues, which is induced by a variety of external cues, including growth-promoting factors.In the nervous system, ERK phosphorylation is induced by binding of neurotrophins to their specific tyrosine kinase receptors or by neuronal activity leading to glutamate release and binding to its ionotropic and metabotropic receptors. Some studies have provided evidence of its importance in neuroplastic events. In particular, ERK phosphorylation in the spinal cord was shown to be nociceptive-specific and its upregulation, occurring in cases of chronic inflammatory and neuropathic pain, seems to be of the utmost importance to behavioural changes observed in those conditions. In fact, experiments using specific inhibitors of ERK phosphorylation have proved that ERK directly contributes to allodynia and hyperalgesia caused by spinal cord injury or chronic pain. Additionally, spinal ERK phosphorylation regulates the micturition reflex in experimental models of bladder inflammation and chronic spinal cord transection.In this review we will address the main findings that suggest that ERK might be a future therapeutic target to treat pain and other complications arising from chronic pain or neuronal injury. PMID- 19305742 TI - Pharmacology of cell adhesion molecules of the nervous system. AB - Cell adhesion molecules (CAMs) play a pivotal role in the development and maintenance of the nervous system under normal conditions. They also are involved in numerous pathological processes such as inflammation, degenerative disorders, and cancer, making them attractive targets for drug development. The majority of CAMs are signal transducing receptors. CAM-induced intracellular signalling is triggered via homophilic (CAM-CAM) and heterophilic (CAM - other counter receptors) interactions, which both can be targeted pharmacologically. We here describe the progress in the CAM pharmacology focusing on cadherins and CAMs of the immunoglobulin (Ig) superfamily, such as NCAM and L1. Structural basis of CAM mediated cell adhesion and CAM-induced signalling are outlined. Different pharmacological approaches to study functions of CAMs are presented including the use of specific antibodies, recombinant proteins, and synthetic peptides. We also discuss how unravelling of the 3D structure of CAMs provides novel pharmacological tools for dissection of CAM-induced signalling pathways and offers therapeutic opportunities for a range of neurological disorders. PMID- 19305743 TI - Endocannabinoid signaling in midbrain dopamine neurons: more than physiology? AB - Different classes of neurons in the CNS utilize endogenous cannabinoids as retrograde messengers to shape afferent activity in a short- and long-lasting fashion. Transient suppression of excitation and inhibition as well as long-term depression or potentiation in many brain regions require endocannabinoids to be released by the postsynaptic neurons and activate presynaptic CB1 receptors. Memory consolidation and/or extinction and habit forming have been suggested as the potential behavioral consequences of endocannabinoid-mediated synaptic modulation. HOWEVER, ENDOCANNABINOIDS HAVE A DUAL ROLE: beyond a physiological modulation of synaptic functions, they have been demonstrated to participate in the mechanisms of neuronal protection under circumstances involving excessive excitatory drive, glutamate excitotoxicity, hypoxia-ischemia, which are key features of several neurodegenerative disorders. In this framework, the recent discovery that the endocannabinoid 2-arachidonoyl-glycerol is released by midbrain dopaminergic neurons, under both physiological synaptic activity to modulate afferent inputs and pathological conditions such as ischemia, is particularly interesting for the possible implication of these molecules in brain functions and dysfunctions. Since dopamine dysfunctions underlie diverse neuropsychiatric disorders including schizophrenia, psychoses, and drug addiction, the importance of better understanding the correlation between an unbalanced endocannabinoid signal and the dopamine system is even greater. Additionally, we will review the evidence of the involvement of the endocannabinoid system in the pathogenesis of Parkinson's disease, where neuroprotective actions of cannabinoid-acting compounds may prove beneficial.The modulation of the endocannabinoid system by pharmacological agents is a valuable target in protection of dopamine neurons against functional abnormalities as well as against their neurodegeneration. PMID- 19305746 TI - Could MRI and CT Scanners Be Operated More Intensively in Canada? AB - Although availability of necessary equipment could play a role in wait times for MRI and CT exams in Canada, there are other dimensions to this issue. More machines in operation do not necessarily reduce wait times. It is important also to consider the level of utilization of the existing pool of scanners.This paper analyzes utilization of MRI and CT scanners (machines) by focusing on two indicators: the number of exams per machine per year and the number of hours of operation per machine per week. These values were calculated and reported by province, followed by an assessment of the average level of utilization of MRI and CT scanners in Canada. The findings suggest that some provinces use their MRI or CT scanners less intensively than others. On average, in Canada, an additional 31% operating capacity may exist for MRI and 68% for CT without additional capital or infrastructure investments. However, supply-side as well as demand side constraints may prevent a given jurisdiction from operating at full capacity. PMID- 19305745 TI - The neuropharmacology of (-)-stepholidine and its potential applications. AB - (-)-Stepholidine (SPD), a natural product isolated from the Chinese herb Stephania, possesses dopamine (DA) D1 partial agonistic and D2 antagonistic properties in the nigrostriatal and mesocorticolimbic DAergic pathways. These unique dual effects have suggested that SPD can effectively restore previously imbalanced functional linkage between D1 and D2 receptors under schizophrenic conditions, in which, SPD improves both the negative and positive symptoms of schizophrenia. SPD also relieves the motor symptoms of Parkinson's disease (PD) when co-administered with Levodopa. Furthermore, SPD exhibits neuroprotective effects through an antioxidative mechanism and slows down the progression of neuronal degeneration in the substantia nigra (SN) of PD patients and/or animal models. Therefore, SPD is a novel, natural compound with potentially therapeutic roles in the treatment of schizophrenia and/or PD. PMID- 19305744 TI - Emerging synergisms between drugs and physiologically-patterned weak magnetic fields: implications for neuropharmacology and the human population in the twenty first century. AB - Synergisms between pharmacological agents and endogenous neurotransmitters are familiar and frequent. The present review describes the experimental evidence for interactions between neuropharmacological compounds and the classes of weak magnetic fields that might be encountered in our daily environments. Whereas drugs mediate their effects through specific spatial (molecular) structures, magnetic fields mediate their effects through specific temporal patterns. Very weak (microT range) physiologically-patterned magnetic fields synergistically interact with drugs to strongly potentiate effects that have classically involved opiate, cholinergic, dopaminergic, serotonergic, and nitric oxide pathways. The combinations of the appropriately patterned magnetic fields and specific drugs can evoke changes that are several times larger than those evoked by the drugs alone. These novel synergisms provide a challenge for a future within an electromagnetic, technological world. They may also reveal fundamental, common physical mechanisms by which magnetic fields and chemical reactions affect the organism from the level of fundamental particles to the entire living system. PMID- 19305747 TI - Influencing Drug Prices through Formulary-Based Policies: Lessons from New Zealand. AB - A national formulary has been proposed as a priority element of Canada's National Pharmaceuticals Strategy. We review a variety of formulary-based policies that might be used in conjunction with a national formulary, drawing on the policies and practices of the Pharmaceutical Management Agency of New Zealand. We consider the potential price impact of an actively managed national formulary by conducting a Canada-New Zealand price comparison for equivalent products in the four largest drug classes: statins, angiotensin-coverting enzyme (ACE) inhibitors, selective serotonin reuptake inhibitors (SSRIs) and proton pump inhibitors (PPIs). The results suggest that potential price savings for Canada in these drug classes are on the order of 21% to 79%. Such price differences would translate into billions of dollars in annual savings if applied across Canada, potentially offsetting the costs of the expansion of pharmacare coverage necessary to achieve both equity and efficiency goals in this sector. PMID- 19305748 TI - The Magnitude, Share and Determinants of Private Costs Incurred by Clients (and Their Caregivers) of In-home Publicly Financed Care. AB - Home-based health services remain one of the fastest-growing sectors in the Canadian healthcare system. While there have been studies addressing the characteristics of home care users and the determinants of utilization, the costs associated with the use of home care services, particularly private costs, have been largely neglected. To gain a comprehensive appreciation of the financing context in which ambulatory and home-based care is delivered and received, it is imperative to assess costs incurred by clients and their family/friends. Accordingly, this study examined the magnitude and determinants of the share of private costs incurred by Ontarians who received in-home publicly financed services and by their unpaid caregivers. The private share of costs was found to be 75%. Determinants of the private share included participants' gender, martial status, functioning in activities of daily living and the type and length of service received. These findings suggest that recipients of home-based health services in Ontario may bear an economic burden when care is shifted into the home setting. PMID- 19305749 TI - Integrating public health and primary care. AB - PURPOSE: Improved health and social outcomes would be possible with better coordination and collaboration between public health and primary care. The purpose of this study is to identify linkages between these health sectors with the aim of informing a forward-looking policy approach to integrate public health functions in primary care. METHODS: We searched national and international journals and the grey literature for relevant papers and reports published from January 1999 to December 2003. The final set of documents provided broad coverage of the topic, with emphasis on national and international representation and a special focus on disease surveillance, health promotion, accident and illness prevention and chronic diseases. RESULTS: Three main findings emerged from this study. First, there is a need to understand and clearly articulate the roles and functions of public health and primary care in Canada. Second, the main areas of overlap between these sectors are health surveillance, health promotion and prevention of disease and injury. Third, based on an international literature search, we identified 10 models that demonstrate how these sectors can be integrated; five of them were developed in Canada. CONCLUSIONS: National and international evidence and a variety of working models support the integration of public health functions in primary care. Canada has been a leader in developing models of integrated health systems that combine individualized approaches to influence personal health behaviour and community approaches to influence the health of the population. These integration models could be further developed through a focus on the common need of primary care and public health to address the health implications of the ever-present risk of emerging infectious diseases in Canada. PMID- 19305750 TI - Pharmaceutical use and outcomes: always a need for a sober second look. PMID- 19305751 TI - Plastic brains. AB - Memories fade, alas, and more rapidly with age, though the aging brain holds more tenaciously to the longer past. Or does it? The brain may be continually editing those seemingly clear memories. That the immature brain constructs, "sculpts" itself by configuring its neural linkages to make best use of the sensory input received in early life has been known for decades. The more recent news is that mature brains also re-arrange these linkages as sensory inputs change. So what? Well, by some estimates inadequate stimulation in early childhood leaves 25% of Canadians neurally challenged by the modern world. Countries with systematic early child development programs show better results. Are there also opportunities for exploiting the plasticity of adult brains? (Or is that already happening, all around us?). PMID- 19305752 TI - Ou sont les chercheurs? Speaking at cross-purposes or across boundaries? AB - Knowledge transfer and exchange (KTE) relates to both the translation and transfer of information, as well as the exchange of information, between researchers and decision-makers. Despite recent advances, KTE efforts may be compromised on two fronts: first, the existing reward structure for university based researchers may not be compatible with applied research; and second, there appears to be a lack of research capacity in healthcare organizations. In this short paper, we contest the first of these points, suggesting that applied research can and should be published in high-index journals, and thus the tenure and promotions process does not need reform. Regarding the second point, we suggest that partnerships be formed across healthcare organizations, universities, government agencies and research funders to support the positioning of PhD-trained researchers directly in healthcare delivery organizations. In our view, it is here, once organizational boundaries are crossed, that significant progress will be made in completing the health policy and health research cycles. PMID- 19305753 TI - Commentary: Speaking at Cross-Purposes or across Boundaries? PMID- 19305754 TI - Commentary: Complementary Perspectives on "Speaking at Cross-Purposes or across Boundaries". PMID- 19305755 TI - Same question, different data source, different answers? Data source agreement for surgical procedures on women with breast cancer. AB - This study assessed the accuracy of the Manitoba Cancer Registry (MCR) and two administrative data sources, the Manitoba Health hospital discharge file and the Manitoba Health medical claims file, for capturing surgical procedures related to the treatment of breast cancer. The study cohort included all women diagnosed in Manitoba with invasive or in situ breast cancer between 1995 and 1999. The surgical procedures of interest were mastectomy, breast conserving surgery and axillary node dissection. Analysis focused on assessing concordance between data sources following record linkage. Agreement was measured using the kappa statistic, and chart reviews of discordant information were completed to identify the more reliable data source and to validate data files. The effect of using each data set alone to calculate procedure rates was determined to identify any clinically important differences arising from the choice of data source. Results indicate that capture of breast cancer patients using administrative data sets alone can be quite good and that the population-based cancer registry is superior to other administrative data sets for capturing surgical treatment information on cancer cases. PMID- 19305756 TI - Costs of new atypical antipsychotic agents for schizophrenia: does unrestricted access reduce hospital utilization? AB - OBJECTIVES: To determine whether increased expenditures to provide unrestricted access to expensive atypical antipsychotic drugs would be associated with a reduction in hospital utilization and cost by patients with schizophrenia. Secondary objectives were to determine the factors associated with length of stay (LOS) and readmission for schizophrenia sufferers. METHODS: Retrospective chart review identified all acute hospitalizations for schizophrenia in the province of Newfoundland and Labrador. LOS and readmission rates, as well as risk factors influencing them, were measured during three time periods: (1) 1995/96, beginning of restricted access; (2) 1998, last year of restricted access; and (3) 2000, second year of open access. Average per diem costs were multiplied by LOS to determine hospital expenditures, and the provincial drug plan database provided the amount of money reimbursed for antipsychotic drugs. RESULTS: Days of hospitalization for schizophrenia totalled 15,089 in 1995/96, 16,318 in 1998 and 15,691 in 2000, resulting in per annum costs of $6,474,095, $7,080,065 and $6,615,795, respectively. There were 57 (18.2%) fewer patients hospitalized and 98 (16.7%) fewer admissions during open access (2000) when compared to a period of restricted access (1995/96). However, median LOS in 2000 was significantly longer than in 1995/96 (22.0 vs. 15.0 days, P<0.001), and was independent of other factors significantly associated with LOS (e.g., suicidal ideation on admission). No change in the number of readmissions was observed. Government expenditures for atypical agents were $217,273 in 1995/96 and $3.8 million in 2000, a 17.5-fold increase. CONCLUSIONS: The unrestricted reimbursement policy for atypical antipsychotics was associated with a large increase in drug plan expenditure, which was not offset by a decrease in hospital utilization by schizophrenia sufferers. Although a decrease in hospital admissions occurred, any associated savings were negated by an increase in LOS. PMID- 19305757 TI - Turning vision into reality: successful integration of primary healthcare in taber, Canada. AB - Primary healthcare offers significant benefits to Canadians and to the healthcare system as a whole. The Taber Integrated Primary Healthcare Project (TIPHP) was a three-year primary healthcare renewal initiative involving rural physicians and the Chinook Health Region in Taber, Alberta, Canada. The goal of the project was to improve healthcare services delivery through integration of the services provided by the physician group and the health region in one rural community. Four main enablers emerged as fundamental to the integration process: community assessment and shared planning; evidence-based, interdisciplinary care; an integrated electronic information system; and investment in processes and structures that support change.The outcome of the project has been the implementation of a new model of healthcare delivery that embraces an integrated collaborative team approach in delivering population-based, primary healthcare. Importantly, the TIPHP has influenced regional healthcare policy related to primary healthcare renewal strategies and partnerships. PMID- 19305758 TI - Education level, income level and mental health services use in Canada: associations and policy implications. AB - BACKGROUND: Investigations of socio-economic gradients in mental health services use in Canada have used different measures of socio-economic status and have shown conflicting results. We explored the relationships between education level, income level and mental health services use among people with a mental illness using data from the Canadian Community Health Survey: Cycle 1.2. METHODS: We included adults who met the criteria for an anxiety or depressive disorder in the past 12 months (n=3,101). We calculated the likelihood of seeking mental healthcare from a psychiatrist, psychologist, family physician or social worker over a period of 12 months by education level. RESULTS: For each additional level of education, individuals were 15% more likely to see a psychiatrist, 12% more likely to see a family doctor, 16% more likely to see a psychologist and 16% more likely to see a social worker. DISCUSSION/CONCLUSION: We found marked inequity in mental health services use by education level that was consistent across service types. Programs aiming to deliver targeted services to consumers who have not completed high school should be developed and evaluated. PMID- 19305759 TI - Using health technology assessment to identify research gaps: an unexploited resource for increasing the value of clinical research. AB - Health technology assessments (HTAs) are an as yet unexploited source of comprehensive, systematically generated information that could be used by research funding agencies to formulate researchable questions that are relevant to decision-makers. We describe a process that was developed for distilling evidence gaps identified in HTAs into researchable questions that a provincial research funding agency can use to inform its research agenda. The challenges of moving forward with this initiative are discussed. Using HTA results to identify research gaps will allow funding agencies to reconcile the different agendas of researchers who conduct clinical trials and healthcare decision-makers, and will likely result in more balanced funding of pragmatic and explanatory trials. This initiative may require a significant cultural shift from the current, mostly reactive, funding environment based on an application-driven, competitive approach to allocating scarce research resources to a more collaborative, contractual one that is proactive, targeted and outcomes-based. PMID- 19305760 TI - The effect of evidence-based drug coverage policies on pharmaceutical r&d: a case study from british columbia. AB - BACKGROUND: To manage public expenditures in the mid-1990s, British Columbia implemented evidence-based drug coverage policies, including "reference pricing." Industry lobbied against the province's policy, arguing that reference pricing harms patients and that it is inconsistent with federal and provincial legislation. Researchers and the courts have studied and rejected industry's claims. However, industry also threatened to halt R&D investment in British Columbia and continues to so threaten other provinces contemplating evidence based drug coverage policies. The purpose of this study is to review evidence regarding these threats. METHODS: Provincial-level R&D data for 1988-2006 were used to analyze the impact of BC PharmaCare's policies on pharmaceutical R&D in British Columbia. We used statistical analyses to determine whether the province's policies affected BC-based R&D as expressed in two ways: (1) as inflation-adjusted expenditure per capita in British Columbia and (2) as the ratio of expenditure per capita in the province to expenditure per capita in the rest of Canada. RESULTS: Evidence-based drug coverage policies had no statistically significant negative effects on BC-based pharmaceutical R&D. BC R&D was slightly above expected trends in 1997 and slightly below expected trends in 1998 and 1999 (though not statistically significantly in either case). From 2001 to 2003, BC R&D was (statistically significantly) above expected trends. CONCLUSIONS: While they are part of the politics of the pharmaceutical sector, claims and threats regarding connections between coverage policy and location of R&D investment are not borne out in British Columbia's experience. This is likely because, as suggested by business and economic literature, firms locate R&D based on the expected cost-to-firm and productivity of the R&D investment itself. Prudent policy would therefore manage pharmaceutical expenditures using evidence based policies and pursue scientific and economic development goals through direct and strategic government investment in local scientific capacity. PMID- 19305761 TI - Improving use of medicines for older people in long-term care: contrasting the policy approach of four countries. AB - The quality of nursing home care for older people, including medication use and related outcomes, has been problematic in a number of developed countries. This paper compares the policy approaches to drug prescribing and administration in nursing homes adopted by four countries. The United States has led the way in terms of regulating and inspecting nursing homes, with strict requirements for prescribing psycho-tropic medications, commonly known as "chemical restraints." These requirements have been facilitated by detailed data collection mandated by the US government. Although regulation has led to marked reductions in the prescribing of these agents, underused medications have received little attention. Despite similar problems with the use of psychotropic drugs, the United Kingdom, Australia and New Zealand have adopted a more generic approach to drug use in the nursing home setting, a situation that may reflect the different organization and ethos of healthcare systems in these countries. Developments in systematic medication data capture, greater collaboration and more educational feedback to prescribers and facilities would represent a major step forward in long-term care policy in these latter three countries, while a broader educational focus would further support improvements in the US setting. PMID- 19305763 TI - Sequestered evidence: inaccessible findings from health services and policy research. PMID- 19305762 TI - How Consumerist Do People Want to Be? Preferred Role in Decision-Making of Individuals with HIV/AIDS. AB - BACKGROUND: North American bioethics emphasizes autonomy, but do care recipients want an autonomous role in treatment decision-making? PARTICIPANTS: We surveyed people living with HIV/AIDS (PHAs) treated at 12 clinics affiliated with the HIV Ontario Observational Database (HOOD). DESIGN: The clinics distributed 809 surveys. Demographic information was merged from the HOOD database. MEASUREMENTS: The survey included questions about preferred role, satisfaction with role, trust in physicians, and use and helpfulness of information sources. RESULTS: The 431 responses (response rate 53.3%) reflected the group believed most consumerist (well-educated gay men). Most (87%) had high/moderate trust in physicians and were satisfied with their level of involvement in treatment decision-making. They sought information from many sources (mean 8.2), particularly health providers, but also the Internet (used by 42%, seen as somewhat helpful). Nonetheless, only one respondent (0.2%) was categorized as autonomous/consumerist; 20.9% were passive, while 78.9% wanted a shared role. CONCLUSIONS: Consumerist rhetoric, with its emphasis on patient autonomy, is oversimplified. Even in this knowledgeable population, respondents wanted to understand their disease and options, but not to take on the provider's role. To our respondents, the optimal doctor-patient relationship is best characterized by a shared, trusting relationship between informed patients and expert providers. PMID- 19305764 TI - Richard III, Barer-Stoddart and the Daughter of Time. AB - "Truth is the daughter of Time," said mystery writer Josephine Tey. This point, illustrated in her rehabilitation of the "villainous" King Richard III, is equally apt for a reconsideration of the 1991 Barer-Stoddart report on medical personnel. Canadian physicians have reviled these authors for "creating" a physician shortage by encouraging provincial cuts to medical school enrolment. Yet, data pre- and post-1991 are quite clear: their report did not and could not have had this effect. The physician-to-population ratio has been stable since 1989. Average physician hours of work have fallen, but per capita expenditures on physicians' services (inflation-adjusted) are rising rapidly. A flood of physicians from the major expansion of enrolments now in place threatens serious fiscal trouble over the next two decades, and is likely to pre-empt any significant system reform. PMID- 19305765 TI - Why examining the desirability of health technology matters. AB - Although technology is ubiquitous in healthcare, its impact on people's perceptions and lives is poorly understood. Fresh insights are required to meet current and future technology-related policy challenges. Keeping a population healthy requires considering not only technologies that are used in clinical settings (diagnostic, therapeutic, palliative), but also those used in the community (home care, self-care, technical aids) and those that affect health more broadly (health promotion technologies, occupational health technologies). At the policy making level, understanding the desirability of health technology may prove to be more important than simply appraising its affordability. PMID- 19305766 TI - A general method for identifying excess revisit rates: the case of hypertension. AB - OBJECTIVE: To provide a description and application of a novel methodology for comparing actual to expected visit rates at the physician level (controlling for patient characteristics) that could be employed in healthcare monitoring and management. DATA SOURCES/STUDY SETTING: Two fiscal years (1997/1998 and 1998/1999) of health utilization data extracted from linked administrative data sets on a population-based cohort of 13,688 patients (aged 25+ with hypertension) involving 157 physicians. STUDY DESIGN: We re-analyzed data from a previously published retrospective cohort study to develop and apply a new methodology for identifying higher or lower than expected physician visit rates for hypertension. DATA COLLECTION/EXTRACTION METHODS: We matched each study physician's hypertensive patients on the basis of age, sex, income and co-morbidity to an equal number of control patients drawn from the cohort. We then compared visit rates between the actual practice and the matched control practice. PRINCIPAL FINDINGS: Although the correlation between the visit rates of the two groups of practices was high (r=.87), there were notable differences in rates, suggesting substantial discretionary practice among physicians. CONCLUSIONS: The methodology outlined in this paper provides a basis for identifying variations in visit levels related to discretionary practice patterns and patient preferences. Deviation from expected visit rates provides a potentially useful measure for performance feedback and quality improvement activities. PMID- 19305767 TI - Engaging front-line staff: how a long-term care home is using evidence to build a quality improvement culture. AB - St. Peter's Residence at Chedoke in Hamilton, Ontario, a 210-bed long-term care facility, is building the capacity of front-line employees to become engaged in quality improvement. With training and tools, teams made up of front-line and other staff are becoming engaged in creating a quality improvement culture. This innovative initiative was recently featured in Promising Practices in Research Use, a series produced by the Canadian Health Services Research Foundation highlighting organizations that have invested their time, energy and resources to improve their ability to use research in the delivery of health services. Tell the Foundation your own stories and visit the Promising Practices inventory at http://www.chsrf.ca/promising/index_e.php. PMID- 19305768 TI - The effect of evidence-based drug coverage policies on pharmaceutical r&d: a case study from british columbia. AB - BACKGROUND: To manage public expenditures in the mid-1990s, British Columbia implemented evidence-based drug coverage policies, including "reference pricing." Industry lobbied against the province's policy, arguing that reference pricing harms patients and that it is inconsistent with federal and provincial legislation. Researchers and the courts have studied and rejected industry's claims. However, industry also threatened to halt R&D investment in British Columbia and continues to so threaten other provinces contemplating evidence based drug coverage policies. The purpose of this study is to review evidence regarding these threats. METHODS: Provincial-level R&D data for 1988-2006 were used to analyze the impact of BC PharmaCare's policies on pharmaceutical R&D in British Columbia. We used statistical analyses to determine whether the province's policies affected BC-based R&D as expressed in two ways: (1) as inflation-adjusted expenditure per capita in British Columbia and (2) as the ratio of expenditure per capita in the province to expenditure per capita in the rest of Canada. RESULTS: Evidence-based drug coverage policies had no statistically significant negative effects on BC-based pharmaceutical R&D. BC R&D was slightly above expected trends in 1997 and slightly below expected trends in 1998 and 1999 (though not statistically significantly in either case). From 2001 to 2003, BC R&D was (statistically significantly) above expected trends. CONCLUSIONS: While they are part of the politics of the pharmaceutical sector, claims and threats regarding connections between coverage policy and location of R&D investment are not borne out in British Columbia's experience. This is likely because, as suggested by business and economic literature, firms locate R&D based on the expected cost-to-firm and productivity of the R&D investment itself. Prudent policy would therefore manage pharmaceutical expenditures using evidence based policies and pursue scientific and economic development goals through direct and strategic government investment in local scientific capacity. PMID- 19305769 TI - Emergency Planning in Ontario's Acute Care Hospitals: A Survey of Board Chairs. AB - BACKGROUND: Effective hospital governance depends on proactive board leadership to minimize risk. STUDY AIM: To survey hospital board chairs about governance practices, particularly with respect to approval processes for oversight of management preparedness for unforeseen emergencies. METHODS: A 2004 survey of hospital managers initially suggested greater board leadership in risk management as a desired strategic priority for Ontario's acute care hospitals. Our literature review and panel process defined 34 best practices in board governance, including two practices explicitly addressing the board's role in preparing for risk. RESULTS: Our findings revealed that some boards may not be actively engaged in ensuring that adequate processes are in place to protect against risk. More than one-quarter (n=28, 26.9%) of board chairs reported that they had not approved a management plan to address emergencies. Thirty respondents (28.8%) said they had not approved a process to identify, manage and minimize risks to the hospital's sustainability. Forty-seven respondents (45.2%) said they had not approved both of these two processes. A significant association emerged between boards that had approved both risk preparation strategies and boards that had implemented six key governance practices relating to accountability for leadership and stakeholder communication. PMID- 19305770 TI - Reducing Wait Times through Operations Research: Optimizing the Use of Surge Capacity. AB - Widespread public demand for improved access, political pressure for shorter wait times, a stretched workforce, an aging population and overutilized equipment and facilities challenge healthcare leaders to adopt new management approaches. This paper highlights the significant benefits that can be achieved by applying operations research (OR) methods to healthcare management. It shows how queuing theory provides managers with insights into the causes for excessive wait times and the relationship between wait times and capacity. It provides a case study of the use of several OR methods, including Markov decision processes, linear programming and simulation, to optimize the scheduling of patients with multiple priorities. The study shows that by applying this approach, wait time targets can be attained with the judicious use of surge capacity in the form of overtime. It concludes with some policy insights. PMID- 19305771 TI - What Do People Think Is Important about Primary Healthcare? AB - The purpose of this study was to inform quality improvement and performance measurement initiatives in primary healthcare based on the perceptions of British Columbia residents. Key features of care were identified during focus group discussions on important areas in primary healthcare, particularly those that could be improved.Eleven focus groups (n=75) were held. Ninety-six per cent of participants reported that they had a regular primary healthcare provider and had been with that provider for an average of 8.5 years. We conducted a thematic content analysis using a coding scheme based on a logic model for this sector.Analysis revealed the importance of six domains: accessibility (geographic location and timeliness of appointments), continuity, responsiveness, interpersonal communication, technical quality and whole-person care. Although participants discussed accessibility most frequently, domains more often associated with satisfaction were interpersonal communication and continuity. PMID- 19305772 TI - Breaking the deadlock: public health policy coordination as the next step. AB - Recent public health crises have revealed the extent to which coordinated government activity is crucial for ensuring the efficacy of public policies aimed at protecting, maintaining and improving the health of the population. The need for coherent and effective interventions in many areas of human activity always comes up against the challenges related to the division of responsibilities, power and jurisdictions inherent in public administration. The recently initiated renewal of public health structures in Canada opens up new possibilities for public health and could foster better coordination of public health efforts. This paper shows, however, that the eventual broadening of the traditional mandate of Canadian public health to include the social (non-medical) aspects of health and the articulation of healthy public policies requires intervention at the central policy level. We offer practical observations about the need to foster better policy coordination across sectors of governments, with a view to contributing to the emergence of a comprehensive public health policy in Canada. PMID- 19305773 TI - Is There a Tension between Clinical Practice and Reimbursement Policy? The Case of Osteoarthritis Prescribing Practices in Ontario. AB - BACKGROUND: Reimbursement policies, such as those used to manage the public drug program for senior citizens in Ontario, focus on providing access to cost effective drug therapies. These policies may create a dilemma for physicians who want to prescribe a particular drug to a patient, but must factor reimbursement restrictions affecting patient-level access into the prescribing decision. METHODS: Information was collected from 102 physicians about prescriptions given to osteoarthritis patients (n=2,147) aged 65 years or older. Patients' access to prescribed drugs was determined from their insurance coverage and the reimbursement criteria set out in the formulary of the public Ontario Drug Benefit Program (ODBP). Starting from the assumption that physicians would follow published consensus guidelines respecting gastroprotection when prescribing NSAIDs in these at-risk elderly patients, three groups of physicians were identified from the record of their actual prescriptions. Group A physicians (n=14) prescribed non-selective NSAIDs alone to >60% of their patients. Group B physicians (n=26) prescribed an NSAID + gastroprotective agent or a Cox-2 selective NSAID to >70% of their patients. Group C physicians (n=62) were those that fit into neither category. An open-ended question was included in the study questionnaire to elicit physicians' own interpretation of what impact drug coverage had on their prescribing behaviour. RESULTS: No significant differences were found across groups with respect to years or type of practice, or to patient characteristics (LR=3.00, p>.2). Group C physicians were most likely to change their treatment choice in favour of restricted (limited use) drugs when patients met the criteria for reimbursement or had private insurance and therefore did not have to bear the additional cost out-of-pocket (LR=58.5; p<.0001). INTERPRETATION: Most elderly at-risk patients are prescribed NSAIDs according to the prevailing guidelines. We found, however, that 40% of physicians have prescribing behaviour that favours non-evidence-based (Group A) or evidence-based (Group B) prescribing in this clinical setting irrespective of drug coverage. The remaining 60% of physicians appeared to be more responsive in their prescribing behaviour to financial constraints on patients' access to drugs. They also self identified as most likely to change treatment if drug coverage had been different. These results have important implications for equity and quality of patient care. They also confirm that physicians' knowledge, values and self efficacy are key determinants of prescribing behaviour and require further study to better understand how medical education and third-party policies and programs that govern pharmaceutical care are integrated into physicans' decision-making. PMID- 19305774 TI - Improving the Quality and Capacity of Canada's Health Services: Primary Care Physician Perspectives. AB - OBJECTIVE: This study set out to identify the perspectives of family physicians (FP/GPs) on the quality and capacity of the services they provide and of the system in which they work, to assess their responsiveness to potential changes and to determine their suggestions for future directions to enhance primary care services. METHODS: Thematic results from prior focus groups with FP/GPs provided direction for a questionnaire sent to practitioners in the urban study area. Seventy-four questions, most using a five-point Likert scale, were grouped into 10 sections: physician issues (based on themes from the focus groups), access to specialist services, workload, scope of practice, primary care physician networks, interdisciplinary collaborative practice, complexities and challenges of family practice, future directions, comments and demographics. RESULTS: Five hundred and eighty-three FP/GPs were surveyed, and 300 responses (52%) were analyzed for frequencies and comparisons using SPSS. In addition to informative responses to the various survey sections noted above, specific physician suggestions for future directions to improve quality and capacity were identified. These included access to specialists/consultants, teamwork/collaborative practice, access to diagnostics, electronic records/technology, time and remuneration. CONCLUSIONS: The identified suggestions by FP/GPs to enhance the quality and capacity of health services contribute to a framework for policy development at national, provincial/territorial and regional levels and can be used as a reference point for the progress of primary care reform initiatives. PMID- 19305775 TI - Disparities in Healthcare Access and Use: Yackety-yack, Yackety-yack. PMID- 19305776 TI - Can a learning-disabled nation learn healthcare lessons from abroad? AB - Canada's apparent capacity to reform its health system is inversely proportionate to the volume of high-quality reports that document its need to do so. One of the principal causes of this inertia is our unusual preoccupation with the financial sustainability of the public system, despite compelling evidence that this is a fundamental misdiagnosis. The case made here is that we need to declare a moratorium on the sustainability debate, become more adept at learning which features of international systems we can and cannot easily import, and recognize that what ails our system originates in design rather than the laws of nature. PMID- 19305777 TI - Private health insurance in Germany: consequences of a dual system. AB - A variety of financial and non-financial incentives has resulted in a considerable degree of adverse selection against social health insurance in Germany. Enrollees in private health insurance are healthier, have higher incomes and have fewer dependents than enrollees in social health insurance. Adverse selection decreases average premium income and at the same time increases average healthcare expenditures in social health insurance. As a consequence, financial sustainability of the public system declines. Moreover, financial incentives for healthcare providers have led to preferential treatment for privately insured patients in outpatient care. The dual health insurance system in Germany is therefore inequitable as well as inefficient, and cannot be considered a role model for post-Chaoulli Canada. PMID- 19305778 TI - Spiralling Medical Costs: Why Canada Needs NICE Medicine. AB - Healthcare spending in Canada has grown rapidly in recent years, especially for drugs. This paper discusses the causes of the problem and makes policy proposals. Conflicts of interest (COIs) are a frequent occurrence in medical research and lead to bias. Published studies, especially in the area of clinical trials on drugs, are much more likely to produce findings favourable to the drug when funded by the manufacturer. Bias can occur by various means, including inappropriate study design (such as giving a placebo to control subjects rather than an existing drug) and selective publication of results. COIs also frequently occur with clinical practice guidelines. High-priced (particularly new) drugs are often marketed by inappropriate means. Drug costs in Canada could be greatly reduced if doctors prescribed lower-cost alternatives where appropriate (therapeutic substitution). Proposals are made for changes in the regulatory agencies responsible for the approval of drugs, drug marketing and post-marketing surveillance. In addition, a new regulatory agency is proposed that would examine the value of drugs and medical devices in terms of clinical effectiveness and cost-effectiveness. Such an agency would set the rules for therapeutic substitution and would determine which medical interventions can be used based on agreed cost-effectiveness criteria. PMID- 19305779 TI - Retention of international medical graduates following postgraduate medical training in newfoundland and labrador. AB - We linked the Memorial University of Newfoundland (MUN) postgraduate database with Scott's Medical Database to determine 2004 work locations of physicians who started residency training at MUN by 1998 to assess whether international medical graduates (IMGs) are as likely as MUN and other Canadian medical graduates (CMGs) to work in Canada and Newfoundland and Labrador (NL). In 2004, 66.8% of the residents were in Canada (87.8% MUN graduates, 47.3% IMGs, 67.3% CMGs) and 18.8% were in NL (43.2% MUN graduates, 7.9% IMGs, 4.8% CMGs). Compared to MUN medical graduates, IMGs and CMGs were less likely to work in Canada and NL. PMID- 19305780 TI - The role of evidence in public health policy: an example of linkage and exchange in the prevention of scald burns. AB - Is sound evidence sufficient to change public health practice and policy? In this paper, we describe a campaign to reduce scald burns among children based on compelling evidence of the effectiveness of an intervention to reduce hot tap water temperature. We provide an overview of the problem and the evidence to support our efforts, the context for addressing the scald problem and the lessons learned about why the relationship between evidence and change in practice is not straightforward. PMID- 19305781 TI - Turning the tide on chronic disease: how a province is using evidence to build quality improvement capacity. AB - Saskatchewan's Chronic Disease Management Collaborative is a quality improvement model that brings together healthcare providers to learn about, test and share experiences with improvement ideas in diabetes and coronary artery disease care. This innovative initiative was recently featured in Promising Practices, a monthly series produced by the Canadian Health Services Research Foundation highlighting organizations that have invested their time, energy and resources to try to improve their ability to use research in the delivery of health services. The Promising Practices inventory can be found at www.chsrf.ca/promising/index_e.php. PMID- 19305782 TI - Features of primary healthcare clinics associated with patients' utilization of emergency rooms: urban-rural differences. AB - OBJECTIVE: A 2002 survey of primary healthcare sites found that 51% of rural and 33% of urban primary care patients reported using the hospital emergency room (ER) in the last 12 months. We did a secondary analysis to identify urban-rural differences in accessibility-related organizational features that predicted ER use. METHODS: We collected information on clinic organization and physicians' practice profiles from 100 primary healthcare sites across Quebec and 2,725 of their regular patients, who reported on ER use. We used hierarchical logistic regression to identify organizational features that predict the probability of ER use by patients. RESULTS: Patient confidence in rapid access at their clinic decreases ER use (OR=0.73). Rural sites offer fewer walk-in services or on-site medical procedures and less proximity to laboratory and diagnostic services, but paradoxically, rural patients are more confident that their own physician will see them for a sudden illness. Patients from clinics offering a larger range of medical procedures on site have lower ER use (OR=0.92 per procedure). Rural physicians tend to divide their time between hospital and primary care; doing in patient care increases ER use (OR=1.64). DISCUSSION: Decreased ER use is found in patients of clinics organized to enhance responsiveness to acute needs, especially in rural areas. Although the high rates of ER use in rural areas partly reflect problems with the accessibility of primary care clinics, in a resource-scarce context rural hospital ERs may cover both primary care urgent problems and emergencies. PMID- 19305783 TI - How good is good enough? Standards in policy decisions to cover new health technologies. AB - Health technology coverage decisions require reasonable criteria, for example, the requirement that a technology be effective, efficient, legitimate in purpose, acceptable in its effects, safe and so on. The leap from such criteria to decisions requires not only evidence, but also standards. Decision-makers must specify their values, which apply in general, regarding what is "good enough" before they can judge any technology in particular. This paper will do the following: (1) describe the key analytic tasks involved in defining coverage criteria and their standards, (2) identify some of the policy applications of explicit standards to coverage decisions and (3) review the policy uses of such standards, including some challenges they pose. The problem of identifying cost effectiveness standards will be used to illustrate key issues. It is argued that a precedent-based understanding of standards is relevant in the Canadian policy context, where fairness is crucial. Studies of actual decision-making that seek standards inductively have been misguided in their focus on central tendencies to the neglect of outliers (precedents), while deductive analyses and rules of thumb have been ungrounded in prevailing values. PMID- 19305784 TI - Patient and surgeon views on maximum acceptable waiting times for joint replacement. AB - OBJECTIVE: To assess patient and surgeon views on maximum acceptable waiting times (MAWT) for hip and knee replacement, their determinants and their relationship to levels of urgency based on the Western Canada Waiting List Priority Criteria Score (PCS). METHODS: At the decision date for surgery, orthopaedic surgeons assessed consecutive patients with the PCS and MAWT. Patients were surveyed 3-12 months post-surgery for MAWT and potential determinants. RESULTS: The patient sample of 208 was 56% female, mean age 69 years (SD 11). Mean MAWT for patients was 18 weeks (SD 11) and for surgeons, 17 weeks (SD 11). Median MAWT for three levels of urgency (PCS) ranged from 13-17 weeks (patients) and 9-26 weeks (surgeons). Patient MAWT was unrelated to the surgeon-rated measures: MAWT (r=.05) and the PCS (r=-.10). Multiple regression analysis showed that males, knee vs. hip replacement, a longer waiting time and a perception of fairness in regard to waiting time were significant predictors of longer patient MAWT. Knee replacement, a better ability to walk without significant pain and less potential for progression of the disease were significant predictors of longer surgeon MAWT. CONCLUSIONS: Patient and surgeon perspectives on MAWT are important to the development of waiting time benchmarks. Benchmarks based on levels of urgency ensure a more transparent and fair process for waiting time management. Knowledge of determinants of MAWT should inform better management of waiting time and access, by understanding the basis of patient and physician views on acceptable waiting times. PMID- 19305785 TI - GABA(A) receptors in normal development and seizures: friends or foes? AB - GABA(A) receptors have an age-adapted function in the brain. During early development, they mediate excitatory effects resulting in activation of calcium sensitive signaling processes that are important for the differentiation of the brain. In more mature stages of development and in adults, GABA(A) receptors transmit inhibitory signals. The maturation of GABA(A) signaling follows sex specific patterns, which appear to also be important for the sexual differentiation of the brain. The inhibitory effects of GABA(A) receptor activation have been widely exploited in the treatment of conditions where neuronal silencing is necessary. For instance, drugs that target GABA(A) receptors are the mainstay of treatment of seizures. Recent evidence suggests however that the physiology and function of GABA(A) receptors changes in the brain of a subject that has epilepsy or status epilepticus.This review will summarize the physiology of and the developmental factors regulating the signaling and function of GABA(A) receptors; how these may change in the brain that has experienced prior seizures; what are the implications for the age and sex specific treatment of seizures and status epilepticus. Finally, the implications of these changes for the treatment of certain forms of medically refractory epilepsies and status epilepticus will be discussed. PMID- 19305786 TI - ThermoTRP channels in nociceptors: taking a lead from capsaicin receptor TRPV1. AB - Nociceptors with peripheral and central projections express temperature sensitive transient receptor potential (TRP) ion channels, also called thermoTRP's. Chemosensitivity of thermoTRP's to certain natural compounds eliciting pain or exhibiting thermal properties has proven to be a good tool in characterizing these receptors. Capsaicin, a pungent chemical in hot peppers, has assisted in the cloning of the first thermoTRP, TRPV1. This discovery initiated the search for other receptors encoding the response to a wide range of temperatures encountered by the body. Of these, TRPV1 and TRPV2 encode unique modalities of thermal pain when exposed to noxious heat. The ability of TRPA1 to encode noxious cold is presently being debated. The role of TRPV1 in peripheral inflammatory pain and central sensitization during chronic pain is well known. In addition to endogenous agonists, a wide variety of chemical agonists and antagonists have been discovered to activate and inhibit TRPV1. Efforts are underway to determine conditions under which agonist-mediated desensitization of TRPV1 or inhibition by antagonists can produce analgesia. Also, identification of specific second messenger molecules that regulate phosphorylation of TRPV1 has been the focus of intense research, to exploit a broader approach to pain treatment. The search for a role of TRPV2 in pain remains dormant due to the lack of suitable experimental models. However, progress into TRPA1's role in pain has received much attention recently. Another thermoTRP, TRPM8, encoding for the cool sensation and also expressed in nociceptors, has recently been shown to reduce pain via a central mechanism, thus opening a novel strategy for achieving analgesia. The role of other thermoTRP's (TRPV3 and TRPV4) encoding for detection of warm temperatures and expressed in nociceptors cannot be excluded. This review will discuss current knowledge on the role of nociceptor thermoTRPs in pain and therapy and describes the activator and inhibitor molecules known to interact with them and modulate their activity. PMID- 19305787 TI - Alcohol related changes in regulation of NMDA receptor functions. AB - Long-term alcohol exposure may lead to development of alcohol dependence in consequence of altered neurotransmitter functions. Accumulating evidence suggests that the N-methyl-D-aspartate (NMDA) type of glutamate receptors is a particularly important site of ethanol's action. Several studies showed that ethanol potently inhibits NMDA receptors (NMDARs) and prolonged ethanol exposition leads to a compensatory "up-regulation" of NMDAR mediated functions. Therefore, alterations in NMDAR function are supposed to contribute to the development of ethanol tolerance, dependence as well as to the acute and late signs of ethanol withdrawal.A number of publications report alterations in the expression and phosphorylation states of NMDAR subunits, in their interaction with scaffolding proteins or other receptors in consequence of chronic ethanol treatment. Our knowledge on the regulatory processes, which modulate NMDAR functions including factors altering transcription, protein expression and post translational modifications of NMDAR subunits, as well as those influencing their interactions with different regulatory proteins or other downstream signaling elements are incessantly increasing. The aim of this review is to summarize the complex chain of events supposedly playing a role in the up-regulation of NMDAR functions in consequence of chronic ethanol exposure. PMID- 19305788 TI - Pharmacodynamics of memantine: an update. AB - Memantine received marketing authorization from the European Agency for the Evaluation of Medicinal Products (EMEA) for the treatment of moderately severe to severe Alzheimer s disease (AD) in Europe on 17(th) May 2002 and shortly thereafter was also approved by the FDA for use in the same indication in the USA. Memantine is a moderate affinity, uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist with strong voltage-dependency and fast kinetics. Due to this mechanism of action (MOA), there is a wealth of other possible therapeutic indications for memantine and numerous preclinical data in animal models support this assumption. This review is intended to provide an update on preclinical studies on the pharmacodynamics of memantine, with an additional focus on animal models of diseases aside from the approved indication. For most studies prior to 1999, the reader is referred to a previous review [196].In general, since 1999, considerable additional preclinical evidence has accumulated supporting the use of memantine in AD (both symptomatic and neuroprotective). In addition, there has been further confirmation of the MOA of memantine as an uncompetitive NMDA receptor antagonist and essentially no data contradicting our understanding of the benign side effect profile of memantine. PMID- 19305789 TI - Herbal compounds and toxins modulating TRP channels. AB - Although the benefits are sometimes obvious, traditional or herbal medicine is regarded with skepticism, because the mechanism through which plant compounds exert their powers are largely elusive. Recent studies have shown however that many of these plant compounds interact with specific ion channels and thereby modulate the sensing mechanism of the human body. Especially members of the Transient Receptor Potential (TRP) channels have drawn large attention lately as the receptors for plant-derived compounds such as capsaicin and menthol. TRP channels constitute a large and diverse family of channel proteins that can serve as versatile sensors that allow individual cells and entire organisms to detect changes in their environment. For this family, a striking number of empirical views have turned into mechanism-based actions of natural compounds. In this review we will give an overview of herbal compounds and toxins, which modulate TRP channels. PMID- 19305790 TI - Clearing the brain's cobwebs: the role of autophagy in neuroprotection. AB - Protein aggregates or inclusion bodies are common hallmarks of age-related neurodegenerative disorders. Why these aggregates form remains unclear. Equally debated is whether they are toxic, protective, or simple by-products. Increasing evidence, however, supports the notion that in general aggregates confer toxicity and disturb neuronal function by hampering axonal transport, synaptic integrity, transcriptional regulation, and mitochondrial function. Thus, neuroscientists in search of effective treatments to slow neural loss during neurodegeneration have long been interested in finding new ways to clear inclusion bodies. Intriguingly, two studies using conditional neuron-specific gene ablations of autophagy regulators in mice revealed that autophagy loss elicits inclusion body formation and a neurodegenerative cascade.Such studies indicate autophagy may be a built-in defense mechanism to clear the nervous system of inclusion bodies.This new finding has implications for our understanding of aging and neurodegeneration and the development of new therapies. First, we discuss the pathways underlying autophagy and its controversial role in cell death and survival regulation.We then discuss the physiological role of autophagy in the aging process of the nervous system. In the final portion of this review, we discuss the therapeutic promise of inducing autophagy and the potential side effects of such treatments. PMID- 19305792 TI - Antipsychotics for the treatment of behavioral and psychological symptoms of dementia (BPSD). AB - Behavioral and psychological symptoms of dementia (BPSD), i.e. verbal and physical aggression, agitation, psychotic symptoms (hallucinations and delusions), sleep disturbances, oppositional behavior, and wandering, are a common and potentially severe problem complicating dementia. Their prevalence is very high and it is estimated that up to 90% of patients with Alzheimer's disease (AD) may present at least one BPSD. Beside the obvious impact on the quality of life of people with dementia, BPSD are responsible for increased risk of patient institutionalization and increased costs. Furthermore, they are associated with caregivers' stress and depression. Drugs used include antipsychotics, antidepressants, anticonvulsivants, anxiolytics, cholinesterase inhibitors and N methyl-D-aspartate receptor modulators. Among these, the most commonly used are anti-psychotics. These drugs have been used for many decades, but in the last years new compounds have been marketed with the promise of comparable efficacy but less frequent adverse effects (especially extra-pyramidal side effects). Their safety, however, has been challenged by data showing a potential increase in adverse cerebrovascular side effects and mortality. This review will summarize the pathophysiology and neuropharmacology of BPSD, it will describe the characteristics of the anti-psychotics most commonly used focusing on their efficacy and safety in BPSD. PMID- 19305791 TI - Water and ion channels: crucial in the initiation and progression of apoptosis in central nervous system? AB - Programmed cell death (PCD), is a highly regulated and sophisticated cellular mechanism that commits cell to isolated death fate. PCD has been implicated in the pathogenesis of numerous neurodegenerative disorders. Countless molecular events underlie this phenomenon, with each playing a crucial role in death commitment. A precedent event, apoptotic volume decrease (AVD), is ubiquitously observed in various forms of PCD induced by different cellular insults. Under physiological conditions, cells when subjected to osmotic fluctuations will undergo regulatory volume increase/decrease (RVI/RVD) to achieve homeostatic balance with neurons in the brain being additionally protected by the blood-brain barrier. However, during AVD following apoptotic trigger, cell undergoes anistonic shrinkage that involves the loss of water and ions, particularly monovalent ions e.g. K(+), Na(+) and Cl(-). It is worthwhile to concentrate on the molecular implications underlying the loss of these cellular components which posed to be significant and crucial in the successful propagation of the apoptotic signals. Microarray and real-time PCR analyses demonstrated several ion and water channel genes are regulated upon the onset of lactacystin (a proteosomal inhibitor)-mediated apoptosis. A time course study revealed that gene expressions of water and ion channels are being modulated just prior to apoptosis, some of which are aquaporin 4 and 9, potassium channels and chloride channels. In this review, we shall looked into the molecular protein machineries involved in the execution of AVD in the central nervous system (CNS), and focus on the significance of movements of each cellular component in affecting PCD commitment, thus provide some pharmacological advantages in the global apoptotic cell death. PMID- 19305794 TI - TRPV1: a target for next generation analgesics. AB - Transient Receptor Potential Vanilloid 1 (TRPV1) is a Ca(2+) permeant non selective cation channel expressed in a subpopulation of primary afferent neurons. TRPV1 is activated by physical and chemical stimuli. It is critical for the detection of nociceptive and thermal inflammatory pain as revealed by the deletion of the TRPV1 gene. TRPV1 is distributed in the peripheral and central terminals of the sensory neurons and plays a role in initiating action potentials at the nerve terminals and modulating neurotransmitter release at the first sensory synapse, respectively. Distribution of TRPV1 in the nerve terminals innervating blood vessels and in parts of the CNS that are not subjected to temperature range that is required to activate TRPV1 suggests a role beyond a noxious thermal sensor. Presently, TRPV1 is being considered as a target for analgesics through evaluation of different antagonists. Here, we will discuss the distribution and the functions of TRPV1, potential use of its agonists and antagonists as analgesics and highlight the functions that are not related to nociceptive transmission that might lead to adverse effects. PMID- 19305796 TI - The pros of not being competitive. PMID- 19305793 TI - The effects of maternally administered methadone, buprenorphine and naltrexone on offspring: review of human and animal data. AB - Most women using heroin are of reproductive age with major risks for their infants. We review clinical and experimental data on fetal, neonatal and postnatal complications associated with methadone, the current "gold standard", and compare these with more recent, but limited, data on developmental effects of buprenorphine, and naltrexone. Methadone is a micro-opioid receptor agonist and is commonly recommended for treatment of opioid dependence during pregnancy. However, it has undesired outcomes including neonatal abstinence syndrome (NAS). Animal studies also indicate detrimental effects on growth, behaviour, neuroanatomy and biochemistry, and increased perinatal mortality. Buprenorphine is a partial micro-opioid receptor agonist and a kappa-opioid receptor antagonist. Clinical observations suggest that buprenorphine during pregnancy is similar to methadone on developmental measures but is potentially superior in reducing the incidence and prognosis of NAS. However, small animal studies demonstrate that low doses of buprenorphine during pregnancy and lactation lead to changes in offspring behaviour, neuroanatomy and biochemistry. Naltrexone is a non-selective opioid receptor antagonist. Although data are limited, humans treated with oral or sustained-release implantable naltrexone suggest outcomes potentially superior to those with methadone or buprenorphine. However, animal studies using oral or injectable naltrexone have shown developmental changes following exposure during pregnancy and lactation, raising concerns about its use in humans. Animal studies using chronic exposure, equivalent to clinical depot formulations, are required to evaluate safety. While each treatment is likely to have maternal advantages and disadvantages, studies are urgently required to determine which is optimal for offspring in the short and long term. PMID- 19305795 TI - Neurodegenerative diseases of the retina and potential for protection and recovery. AB - Recent advances in our understanding of the mechanisms in the cascade of events resulting in retinal cell death in ocular pathologies like glaucoma, diabetic retinopathy and age-related macular degeneration led to the common descriptive term of neurodegenerative diseases of the retina. The final common pathophysiologic pathway of these diseases includes a particular form of metabolic stress, resulting in an insufficient supply of nutrients to the respective target structures (optic nerve head, retina). During metabolic stress, glutamate is released initiating the death of neurones containing ionotropic glutamate (N-methyl-D-aspartat, NMDA) receptors present on ganglion cells and a specific type of amacrine cells. Experimental studies demonstrate that several drugs reduce or prevent the death of retinal neurones deficient of nutrients. These agents generally block NMDA receptors to prevent the action of glutamate or halt the subsequent pathophysiologic cycle resulting in cell death. The major causes for cell death following activation of NMDA receptors are the influx of calcium and sodium into cells, the generation of free radicals linked to the formation of advanced glycation endproducts (AGEs) and/or advanced lipoxidation endproducts (ALEs) as well as defects in the mitochondrial respiratory chain. Substances preventing these cytotoxic events are considered to be potentially neuroprotective. PMID- 19305797 TI - Allosteric theory: taking therapeutic advantage of the malleable nature of GPCRs. AB - The description of the allosteric modification of receptors to affect changes in their function requires a model that considers the effects of the modulator on both agonist affinity and efficacy. A model is presented which describes changes in affinity in terms of the constant alpha (ratio of affinity in the presence vs the absence of modulator) and also the constant xi (ratio of intrinsic efficacy of the agonist in the presence vs absence of modulator). This allows independent effects of both affinity and efficacy and allows the modeling of any change in the dose-response curve to an agonist after treatment with modulator. Examples are given where this type of model can predict effects of modulators that reduce efficacy but actually increase affinity of agonist (i.e. ifenprodil) and also of modulators that block the action of some agonists (the CXCR4 agonist SDF-1alpha by the antagonist AMD3100) but not others for the same receptor (SDF-1alpha peptide fragments RSVM and ASLW).'All models are wrong...but some are useful...'anonymous environmental scientist. PMID- 19305798 TI - Allosteric modulation of muscarinic acetylcholine receptors. AB - Muscarinic acetylcholine receptors (mAChRs) are prototypical Family A G protein coupled-receptors. The five mAChR subtypes are widespread throughout the periphery and the central nervous system and, accordingly, are widely involved in a variety of both physiological and pathophysiological processes. There currently remains an unmet need for better therapeutic agents that can selectively target a given mAChR subtype to the relative exclusion of others. The main reason for the lack of such selective mAChR ligands is the high sequence homology within the acetylcholine-binding site (orthosteric site) across all mAChRs. However, the mAChRs possess at least one, and likely two, extracellular allosteric binding sites that can recognize small molecule allosteric modulators to regulate the binding and function of orthosteric ligands. Extensive studies of prototypical mAChR modulators, such as gallamine and alcuronium, have provided strong pharmacological evidence, and associated structure-activity relationships (SAR), for a "common" allosteric site on all five mAChRs. These studies are also supported by mutagenesis experiments implicating the second extracellular loop and the interface between the third extracellular loop and the top of transmembrane domain 7 as contributing to the common allosteric site. Other studies are also delineating the pharmacology of a second allosteric site, recognized by compounds such as staurosporine. In addition, allosteric agonists, such as McN-A-343, AC-42 and N-desmethylclozapine, have also been identified. Current challenges to the field include the ability to effectively detect and validate allosteric mechanisms, and to quantify allosteric effects on binding affinity and signaling efficacy to inform allosteric modulator SAR. PMID- 19305799 TI - Allosteric modulators of class B G-protein-coupled receptors. AB - Class B GPCR's are activated by peptide ligands, typically 30-40 amino acid residues, that are involved in major physiological functions such as glucose homeostasis (glucagon and glucagon-like peptide 1), calcium homeostasis and bone turnover (parathyroid hormone and calcitonin), and control of the stress axis (corticotropin-releasing factor). Peptide therapeutics have been developed targeting these receptors but development of nonpeptide ligands, enabling oral administration, has proved challenging. Allosteric modulation of these receptors provides a potential route to developing nonpeptide ligands that inhibit, activate, or potentiate activation of these receptors. Here the known mechanisms of allosteric modulators targeting Class B GPCR's are reviewed, particularly nonpeptide antagonists of the corticotropin-releasing factor 1 receptor and allosteric enhancers of the glucagon-like peptide-1 receptor. Also discussed is the potential for antagonist ligands to operate by competitive inhibition of one of the peptide binding sites, analogous to the Charniere mechanism. These mechanisms are then used to discuss potential strategies and management of pharmacological complexity in the future development of allosteric modulators for Class B GPCR's. PMID- 19305800 TI - Allosteric modulation of the calcium-sensing receptor. AB - The calcium (Ca(2+))-sensing receptor (CaR) belongs to family C of the G-protein coupled receptors (GPCRs). The receptor is activated by physiological levels of Ca(2+) (and Mg(2+)) and positively modulated by a range of proteinogenic L-alpha amino acids. Recently, several synthetic allosteric modulators of the receptor have been developed, which either act as positive modulators (termed calcimimetics) or negative modulators (termed calcilytics). These ligands do not activate the wild-type receptor directly, but rather shift the concentration response curves of Ca(2+) to the left or right, respectively. Like other family C GPCRs, the CaR contains a large amino-terminal domain and a 7-transmembrane domain. Whereas the endogenous ligands for the receptor, Ca(2+), Mg(2+) and the L alpha-amino acids, bind to the amino-terminal domain, most if not all of the synthetic modulators published so far bind to the 7-transmembrane domain. The most prominent physiological function of the CaR is to maintain the extracellular Ca(2+) level in a very tight range via control of secretion of parathyroid hormone (PTH). Influence on e.g. secretion of calcitonin from thyroid C-cells and direct action on the tubule of the kidney also contribute to the control of the extracellular Ca(2+) level. This control over PTH and Ca(2+) levels is partially lost in patients suffering from primary and secondary hyperparathyroidism. The perspectives in CaR as a therapeutic target have been underlined by the recent approval of the calcimimetic cinacalcet for the treatment of certain forms of primary and secondary hyperparathyroidism. Cinacalcet is the first clinically administered allosteric modulator acting on a GPCR, and thus the compound constitutes an important proof-of-concept for future development of allosteric modulators on other GPCR drug targets. PMID- 19305801 TI - Allosteric modulators for mGlu receptors. AB - The metabotropic glutamate receptor family comprises eight subtypes (mGlu1-8) of G-protein coupled receptors. mGlu receptors have a large extracellular domain which acts as recognition domain for the natural agonist glutamate. In contrast to the ionotropic glutamate receptors which mediate the fast excitatory neurotransmission, mGlu receptors have been shown to play a more modulatory role and have been proposed as alternative targets for pharmacological interventions. The potential use of mGluRs as drug targets for various nervous system pathologies such as anxiety, depression, schizophrenia, pain or Parkinson's disease has triggered an intense search for subtype selective modulators and resulted in the identification of numerous novel pharmacological agents capable to modulate the receptor activity through an interaction at an allosteric site located in the transmembrane domain. The present review presents the most recent developments in the identification and the characterization of allosteric modulators for the mGlu receptors. PMID- 19305802 TI - Allosteric modulators of GABA(B) receptors: mechanism of action and therapeutic perspective. AB - gamma-aminobutyric acid (GABA) plays important roles in the central nervous system, acting as a neurotransmitter on both ionotropic ligand-gated Cl(-) channels, and metabotropic G-protein coupled receptors (GPCRs). These two types of receptors called GABA(A) (and C) and GABA(B) are the targets of major therapeutic drugs such as the anxiolytic benzodiazepines, and antispastic drug baclofen (lioresal(R)), respectively. Although the multiplicity of GABA(A) receptors offer a number of possibilities to discover new and more selective drugs, the molecular characterization of the GABA(B) receptor revealed a unique, though complex, heterodimeric GPCR. High throughput screening strategies carried out in pharmaceutical industries, helped identifying new compounds positively modulating the activity of the GABA(B) receptor. These molecules, almost devoid of apparent activity when applied alone, greatly enhance both the potency and efficacy of GABA(B) agonists. As such, in contrast to baclofen that constantly activates the receptor everywhere in the brain, these positive allosteric modulators induce a large increase in GABA(B)-mediated responses only WHERE and WHEN physiologically needed. Such compounds are then well adapted to help GABA to activate its GABA(B) receptors, like benzodiazepines favor GABA(A) receptor activation. In this review, the way of action of these molecules will be presented in light of our actual knowledge of the activation mechanism of the GABA(B) receptor. We will then show that, as expected, these molecules have more pronounced in vivo responses and less side effects than pure agonists, offering new potential therapeutic applications for this new class of GABA(B) ligands. PMID- 19305803 TI - The nuclear transcription factor CREB: involvement in addiction, deletion models and looking forward. AB - Addiction involves complex physiological processes, and is characterised not only by broad phenotypic and behavioural traits, but also by ongoing molecular and cellular adaptations. In recent years, increasingly effective and novel techniques have been developed to unravel the molecular implications of addiction. Increasing evidence has supported a contribution of the nuclear transcription factor CREB in the development of addiction, both in contribution to phenotype and expression in brain regions critical to various aspects of drug seeking behaviour and drug reward. Abstracting from this, models have exploited these data by removing the CREB gene from the developing or developed mouse, to crucially determine its impact upon addiction-related processes. More recent models, however, hold greater promise in unveiling the contribution of CREB to disorders such as addiction. PMID- 19305805 TI - The genetics of development and behavior in Caenorhabditis elegans. AB - The current genetic research on Caenorhabditis elegans and the application of genetic techniques to the analysis of development and behavior in this animal are reviewed. Some aspects of the work are emphasized more than others and this inevitably reflects the author's own interests and prejudices. An effort was made to point out the advantages that C. elegans offers for certain types of investigations and to point out, in general terms, the relevance of this work to other areas of biological research. PMID- 19305804 TI - Effects of nicotine during pregnancy: human and experimental evidence. AB - Prenatal exposure to tobacco smoke is a major risk factor for the newborn, increasing morbidity and even mortality in the neonatal period but also beyond. As nicotine addiction is the factor preventing many women from smoking cessation during pregnancy, nicotine replacement therapy (NRT) has been suggested as a better alternative for the fetus. However, the safety of NRT has not been well documented, and animal studies have in fact pointed to nicotine per se as being responsible for a multitude of these detrimental effects. Nicotine interacts with endogenous acetylcholine receptors in the brain and lung, and exposure during development interferes with normal neurotransmitter function, thus evoking neurodevelopmental abnormalities by disrupting the timing of neurotrophic actions. As exposure to pure nicotine is quite uncommon in pregnant women, very little human data exist aside from the vast literature on prenatal exposure to tobacco smoke.The current review discusses recent findings in humans on effects on the newborn of prenatal exposure to pure nicotine and non-smoke tobacco. It also reviews the neuropharmacological properties of nicotine during gestation and findings in animal experiments that offer explanations on a cellular level for the pathogenesis of such prenatal drug exposure. It is concluded that as findings indicate that functional nAChRs are present very early in neuronal development, and that activation at this stage leads to apoptosis and mitotic abnormalities, a total abstinence from all forms of nicotine should be advised to pregnant women for the entirety of gestation. PMID- 19305806 TI - Interaction of Pratylenchus brachyurus and Gigaspora margarita on cotton. AB - An endomycorrhizal fungus, Gigaspora margarita, was more effective in stimulating the growth of cotton (Gossypium hirsutum) 'Coker 201' at a low fertility level (1.77 gm 10-10-10 N-P-K/pot) than doubling the fertility rate for nonmycorrhizal plants. Gigaspora margarita alone stimulated shoot growth (height, weight, and flower production by 96%, 553%, and 760%, respectively) and root growth (385%) over that of nonmycorrhizal controls at low fertility. Plant development was also stimulated by G. margarita at the high fertility level (3.54 gm 10-10-10 N-P K/pot), but the magnitude of the increase was not as great as that at the low fertility level. Although cotton was a suitable host for Pratylenchus brachyurus, plant development was not retarded by this nematode at either fertility level. In concomitant culture, mycorrhizal-induced plant growth, and sporulation of the endomycorrhizal fungus were not affected by P. brachyurus. Reproduction of P. brachyurus was similar on mycorrhizal and nonmycorrhizal cotton. PMID- 19305807 TI - Drilomermis leioderma n. gen., n. sp. (Mermithidae:Nematoda) parasitizing Cybister fimbriolatus (Say) (Dystiscidae-Coleoptera). AB - The nematode Drilomermis leioderma n. gen., n. sp. (Merrnithidae) is described from larvae of Cybister fimbriolatus (Say) (Dytiscidae: Coleoptera) in Louisiana. Diagnostic characters of the genus Drilomermis are: medium-sized nematodes with the cuticle appearing smooth (lacking cross fibers) under the light microscope, six cephalic papillae, without mouth papillae, six hypodermal cords at midbody, 2 extremely long spicules (longer than 10 times body width at anus) which are separate and parallel (not twisted), an S-shaped vagina, medium-sized amphids located near head papillae, and postparasitic juvenile with a tail appendage. D. leioderma possesses a ventrally displaced mouth, very long vagina, and male genital papillae arranged in 3 double rows in the vicinity of the cloacal opening. Even when containing multiple parasites, about 40% of the hosts sulwived emergence of the memithids and lived several more days. In nature, some of these hosts may be able to continue their development, which is unusual since most mermithid-parasitized hosts die soon after the nematode emerges. PMID- 19305808 TI - Population densities of nematodes under seven tillage regimes. AB - Numbers of plant-parasitic nematodes on corn growing under seven tillage regimes were monitored. Differences among treatments occurred for Helicotylenchus pseudorobustus, Pratylenchus spp., Xiphinema americanum, dorylaimids, and total numbers of nematodes. Except with members of the Tylenchinae, highest densities occurred in no-till ridge plots and lowest numbers occurred in spring- and fall plowed pots. PMID- 19305809 TI - Descriptions of females (Emended), a male, and juveniles of Paralongidorus microlaimus (Nematoda: Longidoridae). AB - The male and juveniles of Paralongidorus microlaimus Siddiqi, 1964, are described and the female description emended. A key to the nominal species of the genus is given. PMID- 19305810 TI - Peroxidase and 6-phosphogluconate dehydrogenase in resistant and susceptible cotton infected by Meloidogyne incognita. AB - Assays of specific activities and electrophoretic separations of multiple forms of 6-phosphogluconate dehydrogenase and peroxidase in cotton resistant and susceptible to Meloidogyne incognita were conducted 6 days after inoculation. Specific activities were greater in infected than in uninfected roots and also were greater in the resistant cultivar, 'Clevewilt 6-3-5,' than in the susceptible culti.var, 'M8.' In uninfected roots, peroxidase activity was greater in Clevewilt roots than in M8 roots, but activity of 6-phosphoglueonate dehydrogenase was the same. Multiple forms of peroxidase and 6-phosphogluconate dehydrogenase were separated and resolved by polyacrylamide gel electrophoresis. These experiments demonstrated the occurrence of altered metabolism upon infection and differences in enzyme activity between resistant and susceptible cultivars. PMID- 19305811 TI - Development of a soil-temperature data base on Meloidogyne arenaria for a simulation model. AB - Models for the development rate and death rate of Meloidogyne arenaria relative to temperature and time were developed to improve the predictive performance of a computer simulator. About 74% of the eggs hatched fairly rapidly, whereas the remainder hatched at a lower rate and appeared to be uninfluenced by temperature shock and root exudates. Death rates of eggs were rapid at extremes of temperature (5 and 36 C) during the first week, because of sensitivity of younger eggs, and then declined and later increased gradually with time. Death rates varied little with time at optimum temperatures for survival. PMID- 19305812 TI - Effects of cultivars, subsoiling, and fumigation on soybean yields and Meloidogyne incognita populations. AB - Application of 1,2-dibromo-3-chloropropane (DBCP) and subsoiling under the row in Tifton sandy loam heavily infested with Meloidogyne incognita increased yields of four soybean cultivars. These cultivars have levels of resistance to M. incognita as follows: 'Hutton', high; 'Essex', intermediate; and 'Davis' and 'Ransom', low. After growing these four cultivars, sub-soiling, and applying DBCP for 2 years in the same plots, the residual effects of these practices on yield of Davis cultivar and populations of M. incognita were studied. Greatest yields of Davis were obtained on plots previously planted to Hunon and Essex and on plots previously treated with DBCP for 2 years. Residual effects of subsoiling on yield were not significant. Data on nematode populations indicated that some residual effects occurred because of cultivars and nematicides. However, root-knot was suppressed only where DBCP was applied the 2 previous years. Although beneficial residual effects occurred, they were not sufficient for maximum soybean production. PMID- 19305813 TI - Anatomical response of grain sorghum roots to Meloidogyne incognita acrita. AB - The cotton root-knot nematode, Meloidogyne incognita acrita, reproduced on the roots of grain sorghum, causing syncytia in the cortex or stele of lateral roots. Giant cells developed either singly with few nuclei or in groups with many nuclei. Giant cells that developed in groups appeared the same as those which developed singly. The pericycle and endodermis were interrupted at the site of nematode invasion. Large areas of these tissues were absent for one-third of the circumference of the stele and extended 1.5 mm longitudinally along the root. In the area where pericycle and endodernris were absent, the parenchyma of the cortex extended to the vascular elements, and abnormal xylem surrounding giant cells extended into the region of the cortex. Root-knot galls appeared on sorghum roots as elongate swellings, discrete knots, or swellings with root proliferation. Galls were not observed on brace roots. PMID- 19305814 TI - Salts and the infectivity of Romanomermis culicivorax. AB - The effects of common inorganic ions found in freshwater on the infectivity of Romanomermis culicivorax and the survival of its host, Culex pipiens, were tested. In general, the median lethal concentrations found for R. culicivorax were greater than the reported median concentrations of these ions in freshwater but less than the reported maximum natural concentrations. The ion toxicity for R. culicivorax (on a molar basis) increased in the following order: sodium < potassium < calcium; aml chloride < carbonate = sulfate < nitrate < nitrite < phosphate. The larvae of C. pipiens were generally 20 to 75 times as tolerant of higher ion concentrations as were the preparasitic stages of R. culicivorax. PMID- 19305815 TI - Criconematinae habitats and Lobocriconema thornei n. sp. (criconematidae: nematoda). AB - A 16.4-ha arca at the Michigan State University Water Quality Research Site was surveyed to obtain information on the habitats and prominence of taxa of the Criconematinae. Fifteen species representing six genera (Macroposthonia, Lobocriconema, Criconema, Crossonema, Nothocriconema, and Xenocriconemella) of this subfamily were recovered from the experimental site. Species occurrence and population densities were evaluated by using prominence and importance values. The Criconematinae was one of the most prominent and important nematode subfamilies recovered from this area. The species successfully inhabited a broad range of woodlot and field vegetations, and soil management groups. Taxa of the Criconematinae were generally more prominent in woodlot than in field vegetations, although with several important exceptions, especially within the genus Macroposthonia. The second-most prominent and important species recovered was an tandescribed species of Lobocriconerna. It is described as Lobocriconema thornei n. sp., including scanning electron micrographs of females, and descriptions of several of the juvenile stages. PMID- 19305816 TI - Early stages of nematode-induced giant-cell formation in roots of Impatiens balsamina. AB - Giant cells induced in roots of Impatiens balsamina by Meloidogyne javanica and Meloidogyne incognita have been examined by light and electron microscopy. The first sign of giant-cell formation was division of cells surrounding a larva. Cell plate alignment appeared to proceed normally, but cytokinesis was unsuccessful and binucleate cells formed subsequently. No wall breakdown was evident then or later. The number of nuclei appeared to increase by repeated mitosis without separation by cytokinesis. Although no holes in walls were observed, wall stubs were found, and mechanisms for their formation are suggested. PMID- 19305817 TI - Ingestion, retention, and transmission of two strains of raspberry ring-spot virus by Longidorus macrosoma. AB - The transmission of two strains of raspberry ringspot virus (RRV) by small numbers of nematodes was compared. A strain of RRV from Scotland (RRV-S), originally found in the field associated with Longidorus elongatus, was transmitted frequently by L. elongatus but only once by L. macrosoma. A strain from England (RRV-E) associated with L. macrosoma in the field was transmitted infrequently by each species of nematode. The reasons why L. macrosoma infected only a small proportion of bait plants with virus were investigated, and it was found that most of the nematodes tested had fed on the source plants and many had ingested virus. Most nematodes exposed to RRV-E or RRV-S had fed on the roots of the bait plants and, when thin sections were examined by electron microscope, had retained particles (thought to be those of the virus) in the region of the anterior odontostyle, Thus, most nematodes seem to have had ample opportunity to transmit virus, and the low frequency of transmission may have been due to a failure of the virus particles to be released from the site of retention or to a lack of infectivity of the virus when L. macrosoma was the vector and Petunia hybrida was the host. PMID- 19305818 TI - Effects of cycloate on development of Heterodera schachtii and growth of three Beta species. AB - Greenhouse tests were set up to evaluate the effects of the herbicide, cycloate (S-ethyl cydohexylethylthiocarbamate), oil development of Heterodera schachtii and growth of three Beta species. Cycloate added to infested soil enhanced cyst development/gm root on B. vulgaris and larvae/gm of root in B. patellaris and B. procumbens at 4, 16, and 16 mug(a.i.)/gm of soil, respectively. Total numbers of nematodes/individual root system decreased because of poor root growth of seedlings in cycloate-amended soil. Penetration and larval development through stage three did occur in the wild Beta species in any treatment. Thus, resistance of B. patellaris and B. pocumbens to development of H. schachtii was not altered by cycloate. Cycloate also retarded growth (P = 0.05) of the sugarbeet cultivars and B. patellaris at 4 mug(a.i.)/gm and B. procumbens at 16 mug(a.i.)/gm of soil. Higher concentrations of nematodes/gm root in plants growing in cycloate-amended soil may be attributed to factors such as fewer roots available for penetration, possible effects of cycloate on egg hatch, greater attraction of nematodes to roots, and increased susceptibility of roots to larval penetration. Suppression of seedling growth in cycloate-amended soil may be attributed in part to higher nematode density and in part to direct root damage from cycloate. PMID- 19305819 TI - The influence of Pratylenchus penetrans on the incidence and severity of verticillium wilt of potato. AB - The influence of Pratylenchus penetrans on the incidence and severity of Verticillium wilt was examined in the potato cultivars 'Kennebec', 'Katahdin', and 'Abnaki'. Single-stem plants were grown in soil maintained at a temperature of 22 +/- 1 C. Axenically cultured nematodes were suspended in water and introduced to the soil, at a rate of ca 5,000/25.4-cm pot, through holes made around each stem. Ten days after infestation with nematodes, conidial suspensions of Verticillium albo-atrum were introduced into the soil at a rate of ca 1,000,000/pot. Among Katahdin plants, the severity of foliar symptoms was increased in the presence of both pathogens 2 and 3 weeks after soil intestation. During the remaining 5 weeks, severity of foliar symptoms was not different between plants infected by both pathogens and those infected by Verticillium alone. Within the wilt-susceptible cultivar Kennebec and the resistant eultivar Abnaki, no effects on foliar symptom severity were observed. When plant heights, shoot weights, and tuber yields were analyzed, a Pratylenchus-Verticillium interaction was not evident within any of the cultivars tested. Nematode populations in roots and rhizosphere were suppressed in Kennebec and Katahdin plants in the presence of Verticillium. PMID- 19305820 TI - Effects of aldicarb and its sulfoxide and sulfone on the biology of Tylenchulus semipenetrans. AB - In laboratory testing, egg hatch of Tylenchulus semipenetrans was stimulated at concentrations of 1 and 10 microg/ml aldicarb solution and inhibited at 50 and 100 microg/ml. Aldicarb was more inhibitory to egg hatch than the aldicarb sulfoxide and the aldicarb sulfone. Inhibition of hatch at the high concentration was associated with delays in the molting processes, lack of larval movement within the egg, and delays in embryonic development. Nematode motility was reduced at 10, 50, and 100 microg/ml of aldicarb and aldicarb sulfoxide solution, and at 50 and 100 microg/ml aldicarb sulfone. Male development was retarded at 10 microg/nrl and almost completely inhibited at 50 and 100 microg/ml of the three chemicals. In greenhouse tests, female development antl reproduction on roots of citrus seedlings were suppressed by aldicarb at rates of 2.6 microg/ml and completely inhibited at 10.6 microg/ml of soil solution during a 50-day experimental period. Under field conditions, there was little systemic movement of aldicarb into roots located outside treated areas. Aldicarb reduced the nematode larvae and the female adult population in the second year after the second treatment. There were no differences in egg hatch and sex ratio of citrus nematodes between treated and nontreated roots. PMID- 19305822 TI - Root-knot nematodes and the process of ageing in plants. AB - Infection of plants by root-knot nematodes is often accompanied by physiological changes characteristic of ageing. Ultra-low tissue luminescence of infected plants indicated oxidation of cell-membrane lipids. Cells with membranes subjected to oxidation lose some of their capacity for water retention. Treating tomato and radish with lidocaine hydrochloride, an inhibitor of lipid oxidation, retarded above-ground symptoms of root-knot nematode infection and of ageing. PMID- 19305823 TI - Oxygen and the Infectivity of Romanomermis culicivorax. AB - The effects on itffectivity by preparasitic Romanomermis culicivorax resulting from four exposure times to four concentrations of oxygen at three temperatures were studied . Low oxygen (1.8 and 3.1 mg /liter) slowed losses of infectivity. Losses of infectivity were similar under aerobic (6 mg /liter) and microaerobic (0.4 mg /liter) conditions for the first 24 h but thereafter were slower in the microaerobic group. N interactions between temperature and oxygen were found over the ranges tested (15-27 C; 0.4-6.0 mg /liter). PMID- 19305824 TI - Hybridization of races of Heterodera glycines. AB - Progeny from single females of four known races of Heterodera glycines Ichinohe were used to establish relatively uniform populations. Single females from these populations were mated with males of other races in all possible combinations to study compatibility and inheritance patterns. When race 1 or 3 was crossed with either race 2 or 4, there was a significant reduction in number of females and a greater number of eggless females than in crosses of races 1 x 3 and 2 x 4. More females matured and fewer were eggless when matings were of the same race. Parasitic capabilities of races 2 and 4 were dominant or partially dominant over those of races 1 and 3, based on parasitism of F hybrids. Segregation patterns were generally similar for reciprocal crosses between races. There appeared to be either one or two major genes segregating for parasitism of 'Pickett' soybean in the different crosses. A hybrid isolate (race 3 x 4) that differed in parasitic capability from the four known races produced as many females on the resistant soybean genotype, PI 90,763, as on the susceptible Lee cultivar. Those data indicate that isolates of H. glycines with a different parasitic capability may develop from gene recombination. PMID- 19305825 TI - Scanning electron microscopy of perineal patterns of three species of Meloidogyne. AB - The perineal pattern of Meloidogyne graminicola is oval, characterized by the anastomosing of cuticular striae converging, and elevating at the tail terminus. A deep lateral incisure forming a low arch traverses the rounded perineal pattern of Meloidogyne hapla. Punctations at the tail region of M. hapla are not observed with the scanning electron microscope. Minute crosslines invaginate with the vulvar lips of Meloidogyne incognita. Punctate indentations around the anal opening and bordering the lateral sides of the vulvar slit of M. incognita are revealed. PMID- 19305826 TI - Effects of nematicides on nematode densities in turf in Connecticut. AB - The plant-parasitic nematodes Criconemoides lobatum, Hoplolaimus tylenchiformis, and Tylenchorhynchus dubius were present in the top 7.5 cm of sod consisting of numerous stolons and fibrous roots. Phenamiphos and 1,2-dibromo-3-chloropropane (DBCP) controlled all three species, whereas ethoprop and oxamyl controlled H. tylenchiformis and T. dubius but not C. lobatum. Benomyl and carbofuran controlled H. tylenchiformis but had poor control of C. lobatum and T. dubius. The effectiveness of carbofuran varied with the type of formulation, being most effective as a quick-release formulation. C. lobatum was the most difficult to control with chemicals. No chemical treatment improved the growth of 'Astoria colonial' bentgrass (A. tenuis Sibth.) or Kentucky bluegrass (Poa pratensis L.) under the moist condtions prevalent in these tests. PMID- 19305827 TI - External morphology of the spicules of some Trichodoridae. AB - External morphology of spicules in several species of Trichodoridae was studied by scanning electron and light microscopy. The bristles on the spicules observed in the light microscope in several species were revealed as small scales forming a sheath which covers most of the spicule body. Some species have smooth spicules, whereas other species exhibited complicated structures formed by projections. In all of the species studied, either a ventro-terminal or terminal opening of the spicules was observed. PMID- 19305828 TI - Three new species of trichodoridae (nematoda: diphtherophorina) with observations on the vulva in Paratrichodorus. AB - Trichodorus intermedius n. sp., Trichodorus dilatatus n. sp., and Paratrichodorus (Atlantadorus) grandis n. sp. are described from soil around native plants of Southern California. The females of the three subgenera of Paratrichodorus are further characterized on the basis of the vulva, which is pore-like in Atlantadorus, a longitudinal slit in Paratrichodorus, and a transverse slit in Nanidorus. PMID- 19305829 TI - Systematics of the monodelphic species of trichodoridae (nematoda: diphtherophorina) with descriptions of a new genus and four new species. AB - To show the relationship of the monodelphic species of Trichodoridae with the nominal taxa, the genus Monotrichodorus is redefined, with the addition of one new species. The new genus Allotrichodorus is proposed on the basis of new species found in Brazil; and one new species is described in the genus Paratrichodorus. PMID- 19305830 TI - Intraspecific morphological variation among populations of Pratylenchus brachyurus and P. coffeae. AB - Three populations of Pratylenchus coffeae and two of P. brachyurus, each originating from a single female, were maintained on Citrus spp. or Solanum nigrum L. for several years under greenhouse conditions. Nematodes were extracted from roots, and adult female specimens were killed, fixed, and mounted in glycerine for microscopic study. Variables measured were distance between vulva and anus and lengths of the stylet, posterior uterine sac, and tail. The mean data and coefficients of variability suggest that styler length had the least variability, and length of posterior uterine sac the most. When males and distinct spermathecae are not evident in P. coffeae populations, the species can he distinguished from P. brachyurus by a shorter mean stylet length, longer mean posterior uterine sac length, and much longer distance between the vulva and anus. PMID- 19305831 TI - Com yield increases relative to nonfumigant chemical control of nematodes. AB - Corn yields were measured after application of nematicides in 16 experiments, mostly in medium-to-heavily textured soil, at 12 locations in Iowa during 1973 1976. The average maximum yield increase in plots treated with nematicides was 21% over yields in untreated plots. Yields were correlated negatively with nematode numbers or nematode biomass in nearly all comparisons. Correlations of nematode numbers in the soil with yield averaged -0.56 for Helicotylenchus pseudorobustus, -0.45 for Hoplolaimus galeatus, -0.51 for Pratylenchus spp., and 0.64 for Xiphinema americanum. Correlation coefficients for numbers of nematodes in the roots and yield averaged -0.63 for Pratylenchus spp. and -0.56 H. galeatus. Correlation coefficients for yield and total number of nematodes averaged -0.65 in roots and -0.55 in soils. Negative correlations also were greater for comparisons of yield with total parasitic-nematode biomass than with numbers of individual nematodes of a species or total numbers of parasitic nematodes. PMID- 19305832 TI - Influence of low temperature on rate of development of Meloidogyne incognita and M. hapla larvae. AB - Development of Meloidogyne incognita and M. hapla larvae in clover roots was studied at 20, 16, 12, and 8 C in growth chambers and in the field from fall through spring, in North Carolina. Larvae of both species invaded roots and developed at 20, 16, and 12 C, but not at 8 C. The time necessary to complete the larval stages at each temperature was determined. The minimal temperature for development of M. incognita larvae was 10.08 C and 8.8 C for M. hapla larvae. In the field, soil temperature at 10 cm deep was favorable for development of larvae until the end of November, and again from February on. All stages of the nematodes survived freezing temperatures in the roots. Reproduction of both species was evident in March or Apri1 after inoculation and accumulation of 8,500 to 11,250 degree-hours. PMID- 19305833 TI - Community analyses of plant-parasitic nematodes in the Kalsow Prairie, Iowa. AB - Twenty-one species of plant-parasitic nematodes were recovered from 15 sites in the Kalsow Prairie, Iowa. Nematode communities were analyzed by prominence and importance values of the nematode species and also by diversity and concentration of dominance. The use of numbers and biomass were compared in indices of diversity and concentration of dominance. Tylenchorhynchus maximns ranked first in mean density/site, prominence value, and importance value, although it was not found as frequently as many other nematodes. Xiphinema americanum and T. maximus were among the dominant nematodes in 11 of 15 sites when biomass was used in the concentration-of-dominance index, but they were dominant in only five sites when numbers were used. PMID- 19305834 TI - Influence of chilling and freezing temperatures on infectivity of Meloidogyne incognita and M. hapla. AB - Egg masses and second-stage larvae of Meloidogyne incognita and M. hapla in soil were exposed to temperatures ranging from 20 to -8 C. Temperature was lowered in 2-day intervals to 16, 12, 8, 4, 0, -4, and -8 C, and the nematodes remained at 4, 0, -4, or -8 C for 18, 14, 10, or 6 days, respectively. Unhatched larvae of both species were more resistant to low temperatures than were embryonic stages. Within the eggs of M. incognita, 7.5% of embryos and 48% of larval stages survived 14 days at 0 C, whereas 9% of embryos and 90% of larval stages in the eggs of M. hapla survived 10 days at -4 C. Second-stage larvae of both species remained infective in sol.1 at 4 or 0 C, but were injured at -4 and -8 C. Infectivily of these larvae was lower in saturated soil than in soil at 51 cm moisture tension at all temperatures. PMID- 19305835 TI - Reaction of citrus rootstocks to Meloidogyne javanica. AB - The response of Citrus spp. and related rootstocks to a population of Meloidogyne javanica was evaluated in a screenhouse experiment. Palestine and Rangpur lime, rough lemon, sour orange, Sexton and Thentriton tangelo, and Volkamer lemon were not infected by M. javanica. Galls and tip swellings were observed on the roots of Poncirus triloliata and Troyer citrange. There was no evidence of nematode development. Symptoms induced by the nematode were stelar division, syncytia formation in the vascular tissues, and necrotic cells. PMID- 19305836 TI - Iontophoretic cobalt staining of the body wall of Phocanema decipiens. AB - Iontophoretic cobalt staining of the nematode Phocanema decipiens results in the deposition of cobalt between the contractile bases of adjacent muscle cells and in a hexagonal lattice pattern in the hypodermis. The possibility of staining sarcolemmal invaginations between muscle cells which are proprioceptive or coordinate the activity of adjacent muscle cells is suggested. PMID- 19305837 TI - Ontogeny of Daucus carota infected with Meloidogyne hapla. AB - The ontogeny of carrots (Daucus carota cv. 'Spartan Premium') grown under greenhouse conditions in pots of organic soil infected with Meloidogyne hapla was influenced detrimentally as early as 4 days after seeding, as determined through analysis of plant surface area, dry weight, fresh weight, net assimilation rate, relative growth rate, and leaf-area ratio. Only 58% of the diseased carrots were suitable for fresh market, compared with 97% of those grown in nematode-free soil. Growth and development of the shoot system (height, surface area, dry weight, and fresh weight) were retarded by M. hapla as early as 12 days after seeding. During the first 12 days after seeding, root dry weight was greater for diseased plants than for controls. Root growth and development (surface area, dry weight, and fresh weight) associated with this nematode, however, were retarded as early as 16 days after seeding. M. hapla caused a delay in the occurrence of 2nd-, 4th-, and 5th-order roots, and an increase in the occurrence of 6th-order roots in infected plants. Parasitized plants had 44% fewer roots (primary through 6th-order) and 50% less total root length. PMID- 19305838 TI - Effects of temperature on the pathogenicity of Tylenchorhynchus clarus to alfalfa and observations on feeding. AB - The involvement of Tylenchorhynchus clarus in plant disease is reported. Addition of a suspension of surface-axenized nematodes reduced top and root growth of alfalfa. Reproduction of T. clarus was greater at 24 and 27 than at 21 C. The interaction of nematodes with temperature did not produce significant effects on alfalfa growth in the 4.5-mo experimental period. T. clarus fed endo- and ectoparasitically. PMID- 19305839 TI - Modification of a computer simulation model for a plant-nematode system. AB - New data on egg development and death rates, and refinements of logic concerning interaction of the nematode and host, were incorporated into a simulation model of a Meloidogyne arenaria and grapevine system. Simulations of field data improved but other areas of weakness in the model were discovered. Two peaks in the egg population curve suggested that the nematode was able to complete two life cycles before host dormancy and declining temperatures limited physiological activity. PMID- 19305840 TI - Dolichodorus aestuarius n. sp. (Nematode: Dolichodoridae). AB - Dolichodorus aestuarius n. sp. from an estuarine habitat near Cedar Key, Florida is described. This nematode has a stylet range of 62-76 microm in females and 60 72 mum in males. The stylet is shorter than those of all described species except D. brevistilus. The probable host plant is Juncus roemerianus. PMID- 19305841 TI - Phytoparasitic nematodes adjacent to established strawberry plantations. AB - Plant-nematode populations associated with uncultivated vegetation, adjacent strawberry plants, and alternate crop sites were studied at three locations in Minnesota. At one site (Forest Lake), Paratylenchus projectus, Meloidogyne hapla, and Pratylenchus tenuis were frequently associated with the roots of native vegetation. These nematode species were also present in adjacent strawberry beds. Among alternate crops observed, oats and muskmelon usually supported the fewest nematodes although moderate densities of Xiphinema americanum and P. tenuis were found at one location in plots planted to oats. Pratylenchus tenuis was also found on rye at one location. PMID- 19305842 TI - The effect of diflubenzuron on egg formation by the root-knot nematode. PMID- 19305843 TI - Influence of Metyrapone on Development of Heterodera glycines. AB - Metyrapone, an inhibitor of steroid synthesis, affected the survival and rate of development of Heterodera glycines. Metyrapone in aqueous tartaric acid solvent influenced sex ratios. The effect on sex ratios was mediated through the host, whereas the effect on survival was apparently effected directly. Tartaric acid increased larval penetration of soybean roots. PMID- 19305844 TI - Movement and Persistence of 1,2-dibromo-3-chloropropane in a Soil with a Plow pan. AB - Movement and persistence of 1,2-dibrotno-3-chloropropane (DBCP) in a Coastal Plain soil containing a sandy plow-pan were enhanced in each of 2 years by subsoiling, increased depth of application, and increased rate of application. DBCP was extracted from the soil with hexane and analyzed by gas chromatography. Subsoiling at a 35-cm depth gave the greatest increase in lateral movement and downward penetration of DBCP in 1975 (a wet year), but bad less effect in 1976 (a dry year). An increased application rate (10 kg/ha vs. 13.5) improved coverage moderately in 1975 by increasing lateral movement, but had little effect in 1976. Increased application depth (18 vs. 35 cm) improved coverage in both years though more in 1976. Deep placement extended DBCP retention time. Rainfall in 1975 probably decreased the number and size of air-filled pores, slowing loss of DBCP to the atmosphere. Because of reduced porosity, the plow-pan was impervious to the passage of DBCP unless disrupted by subsoiling. PMID- 19305845 TI - Resistant Host Responses to Ten California Populations of Meloidogyne incognita. AB - Resistant and susceptible cultivars of tomato, lima beans, cotton, and alfalfa were tested with 10 populations of Meloidogyne incognita from different California locations. Nine of the populations differed in aggressiveness on the nine cultivars tested. Two populations were especially aggressive toward resistant tomato cultivars. PMID- 19305846 TI - Influence of Soil Fumigation on the Fusarium-Root-knot Nematode Disease Complex of Cotton in California. AB - For control of the root-knot nematode, Meloidogyne incognita, and the pathogenic wilt fungus, Fusarium oxysporum, on cotton, soil fumigants were applied in the field at conventional and higher rates. Conventional rates suppressed Fusarium wilt but higher rates gave quicker early growth, better stands, less stand loss over the season, a lower percentage of plants infected with wilt, fewer plants with vascular discoloration, and fewer nematodes. The best treatment about doubled the yields of untreated controls in one experiment and quadrupled them in another. PMID- 19305847 TI - Pathogenicity of Macroposthonia xenoplax to Walnut. AB - Preplanting treatment of soil naturally infested with Macroposthonia xenoplax with 1,2-dibromoethane (ethylene dibromide) significantly increased the growth rate of Juglans hindsii seedlings. When seedlings of J. hindsii, J. regia CV "Serf" and J. regia CV Eureka were inoculated with M. xenoplax, their growth was signilicantly less than that of nematode-free controls. This retarded growth rate was accompanied hy feeder root necrosis, longitudinal cracks in the older roots, and distinct lesions in the secondary phloem. PMID- 19305848 TI - Parasitism of Meloidogyne eggs by a new fungal parasite. AB - Dactylella oviparasitica, a fungus isolated from Meloidogyne egg masses, was shown to parasitize eggs on agar and in soil, when inoculated as either myceliunt or conidia. The fungus also grew saprophytically on eggs killed ,with methyl bromide or heat. The amount of parasitism in the laboratory suggested that the fungus may be a useful biological control agent against Meloidogyne. PMID- 19305849 TI - Infectivity of Neoaplectana carpocapsae and Heterorhabditis heliothidis to pupae of the parasite Apanteles militaris. AB - The infectivity of Neoaplectana carpocapsae and Heterorhabditis heliothidis to Apanteles militaris, a gregarious parasite of the armyworm, was deterntined at 100. 1,000, 5,000, and 10,000 nematodes per petri dish. For both nematode species, the percentage of infected A. militaris within a cocoon cluster decreased as inoculum levels decreased. At the highest inoculum level, N. carpocapsae infected an average of 32% of the parasite pupae within a cocoon cluster, whereas H. heliothidis infected an average of 22%. Covariance analysis indicated, however, that N. carpocapsae had significantly greater infectivity than did H. heliothidis. Some of the dauer juveniles of N. carpocapsae on the body of the armyworm contacted the emerging parasites and eventually became enveloped within the silken cocoons. Dauer juveniles produced by N. carpocapsae in parasite pupae could not penetrate and escape from silken cocoons even when the cocoons were placed in a moist environment. PMID- 19305850 TI - Ontogeny and Identification of Life-Cycle Stages of Criconema octangulare (Cobb, 1914) Taylor, 1936. AB - The ontogeny of Criconema octangulare (Cobb, 1914) Taylor, 1936, is examined and shows ovary development and body length to be of primary importance in differentiating its life-cycle stages. Second-stage juveniles can be identified by body length and ovary length. The third stage can be distinguished from the second by longer body and ovary length, and from the fourth by a shorter ovary. The fourth stage can be identified by stylet length and ovary length. Included is a key to the life stages of C. octangulare, a description of the life-cycle stages, and light and scanning-electron microscopy illustrations. PMID- 19305851 TI - A New Ataloderinae (Nematoda: Heteroderidae), Thecavermiculatus gracililancea n. gen., n. sp. AB - Thecavermiculatus gracililancea n. gen., n . sp. is described from the roots of Festuca myuros L. ("rattail fescue") a range grass in Monterey County, California. Thecavermiculatus females have a slight terminal prominence on which are located the vulva and anus, while its closest relatives, Atalodera and Sherodera, have a large terminal prominence. Furthermore, in T. gracililancea the hatched second-stage juveniles are retained in the female body while Atalodera and Sherodera females retain embryonated eggs. PMID- 19305852 TI - Influence of Population Densities of Heterodera schachtii on Sugar Beets Grown in Microplots. AB - High initial population densities of Heterodera schachtii larvae (36 and 108/gm of soil) greatly retarded the seedling emergence of sugar beet 'Monogerm CSF 1971' in Vineland fine sandy loam. In comparison with controls, initial population densities (P(i)'s) of 1.7, 3.0, 6.2, and 14.4 larvae/gm of soil respectively reduced the weight of storage roots by 38, 56, 64, and 92%. Weights of tops also decreased with increases in P(i); weights of tap and small feeder roots tended to be higher at all P(i)'s except the highest. Sucrose percentage was not affected by any initial nematode density. The populations were lower at midseason than at seeding, and at harvest had increased greatly, with respective populations of 339, 402, 222, and 140 larvae/gm of soil. At harvest, cysts/gm of soil and cysts/gm of root were respectively 4.4 and 72, 6.1 and 99, 6.1 and 191, and 5.8 and 140. The maximum rate of multiplication was 150-200. and maximum density was 400 larvae/gm of soil. The high pathogenicity and multiplication rate of the nematode was attributed to optimum temperature conditions and soil type. PMID- 19305853 TI - Growth response of three vegetables to smooth- and crenate-tailed females of three species of pratylenchus. AB - The effect of morphological variants of females of Pratylenchus penetrans, P. neglectus, and P. crenatus on the growth of three vegetables was studied. Variants were characterized by having either a smooth or crenate tail terminus. Pea was inoculated with variants of P. penetrans, one female per seedling, and grown at light intensities ranging from 1,350 to 21,600 lux in a series of five experiments. Only crenate-tailed females of P. penetrans suppressed the growth of pea and only when pea was grown at 3,900 lux. Radish was inoculated with morphological variants of P. penetrans, P. neglectus, and P. crenatus, four females per seedling, and grown at 3,900 lux in two experiments. Again, truly creuate-tailed females of P. penetrans inhibited growth. The two variants of P. penetrans had a similar infectivity, greater than that of the other two species of Pratylenchus. Only crenate-tailed P. penetrans reproduced on radish. Onion was inoculated with variants of P. penetrans and P. crenatus, four females per seedling, and grown at 14 C at 12,900 lux. Again, only crenate-tailed P. penetrans inhibited growth. The variants of P. penetrans had a similar infectivity, greater than that of P. crenatus. Neither species reproduced on onion at low temperatures. PMID- 19305854 TI - The Sterol, Fatty Acid, and Hydrocarbon Composition of Globodera solanacearum. AB - Globodera solanacearum females were found to have less than 0.01% of dry wt as sterols. Seven sterols were detected in the nematode, with stanols (campestanol and stigmastanol) making tip more than 50% of the total sterols present. Lipid amounted to 29.4% of the dry weight of the nematode. Triglyceride, free fatty acid, and phospholipid classes were composed predominantly of 20:4, 20:1, and 18:1 fatty acids. Of the total weight of fatty acids found in G. solanacearum females, the greatest portion occurred in the triglyceride fraction, followed by the free fatty acid fraction then the phospholipid fraction. Several unidentified hydrocarbons were detected in the nematode. Paraffinic hydrocarbons detected ranged in carbon length from C15 to C29. Total concentration of hydrocarbon composed 0.20% of the dry wt. PMID- 19305855 TI - Meloidogyne incognita wartellei n. subsp. (Meloidogynidae), a Root-knot Nematode on Resistant Soybeans in Louisiana. AB - Meloidogyne incognita wartellei n. subsp, is described and illustrated from roots of soybean (Glycine max L.) near Washington, Louisiana. It is rather limited in distribution in that state, being known at five locations comprising about 60,000 acres. It not only attacks commonly susceptihle soybeans but is a destructive pest on other commercial soybean varieties that are resistant to other forms of the M. incognita group in the area. This new subspecies is related most closely to M. i. incognita and M. i. acrita, but differs especially in the females having a delicate stylet with small, rounded knobs sloping posteriorly; dorsal esophageal gland orific further back (5 mum) from base of stylet; and excretory pore often two to three stylet lengths (sometimes more) from the anterior end. Also, males are often without detectable head annules and with an average stylet length of 22.4 mum. Comments and morphometric data are given on M. i. incognita and M. i. acrita. PMID- 19305856 TI - Measuring Area and Perimeter of Second-Stage Larvae and Males with the Image Analyzing Computer to Distinguish Between Globodera Rostochiensis and G. pallida. PMID- 19305857 TI - Temperature-based Prediction of egg-mass production by Meloidogyne incognita. AB - A maturation-rate relationship for Meloidogyne incognita on Lycopersicon esculentum 'Rutgers' was derived and used to estimate harvest dates for maximum egg hatch from laboratory cultures at ambient temperatures. Daily maturation increments were totaled (nematode maturation total, NMT) and correlated with hatch from isolated white, yellow, and amber egg masses. Hatch per mass fluctuated periodically from ca. 1.0 NMT, when egg masses were first visible, to 2.5 NMT by which time plants showed stress. Maximum yields from white and yellow masses occurred, with a shorter than expected periodicity, at 1.5-1.8 and 2.1-2.2 NMT. White masses decreased from 90% of the total masses at 1.0 NMT to 5% at 2.5 NMT, as the proportion of yellow and amber masses increased concomitantly. Harvested masses per gram of root varied from 97 to 276; hatch per gram of root, 11,000 to 86,000. PMID- 19305858 TI - Evaluation of the protective and therapeutic properties of DBCP for control of root-knot nematode on tomato. AB - Twelve soil drenches over a period of 30 days with DBCP concentrations of 40 microg/ml did not completely prevent infection of tomato plants by root-knot nematode juveniles. Repeated DBCP drenches of 40 microg/ml halted gall development during the drenches, but 10 days after drenching was discontinued galls were apparent. DBCP drenches at 200 microg/ml prevented tomato root development, and 40 microg/ml slowed it. Ten microg/ml increased the height of root-knot-infected plants, but not their top weights. Treated plants were lanky. Protective drenches of 2.5 to 40 microg/ml of DBCP decreased nematode populations and increased fruitfulness. DBCP as a therapeutant reduced the incidence of galling on new roots and halted increases in gall size on previously infected roots but did not improve fruitfulness or plant size significantly. PMID- 19305859 TI - Influence of low temperature on development of Meloidogyne incognita and M. hapla eggs in egg masses. AB - Egg masses of Meloidogyne incognita and M. hapla were placed in soil at 10, 12, 16, and 20 C. At regular intervals, eggs from samples of egg masses were released from the gelatinous matrices and their developmental stages recorded. The number of days necessary to complete each stage from gastrulation to hatch is given for each temperature. The minimal temperature threshold for the development of eggs was computed by linear regression to be 8.26 C for M. incognita and 6.74 C for M. hapla. PMID- 19305860 TI - Influence of potential difference and current on the electrotaxis of Caenorhaditis elegans. AB - C. elegans responds directionally to a DC current. The response may be to the anode or cathode, depending on the current, potential difference, and ion concentration of KC1, Tracks of the responding nematodes show that electrotaxes are genuine orientation phenomena. The directional movement is not due to the passive movement of nematodes or to the influence of currents on the muscular physiology; electrotaxes are mediated sensorily. Details of the response are described. PMID- 19305861 TI - Infectivity of Pratylenchus penetrans on Alfalfa. AB - The infectivity of Pratylenchus penetrans on alfalfa seedlings cv. Du Pulls was studied. The dense root-hair zone was the preferred zone of penetration by females, males, and third-stage larvae. A lesion initially appeared as a water soaked area at the root surface, becoming yellow and elliptical as the nematode entered the cortex, with dark-brown cells later appearing in the centre as the nematode fed. At 20 C, females penetrated roots earlier, faster, and in greater numbers than either males or third-stage larvae. Females penetrated roots at temperatures from 5 to 35 C, with maximum penetration between 10 and 30 C, while males and third-stage larvae penetrated roots only between 10 and 30 C with maximum penetration a t 20 C. Penetration of roots by females, males, and third stage larvae increased after storage of 5 C for 35 days, but decreased after storage of 140 days or more. Combinations of the three life stages in pairs neither enhanced nor inhibited penetration of roots by individual life stages; males were not attracted to females. Increasing inoculum density up to 20 nematodes/seedling did not affect penetration. PMID- 19305862 TI - Molecular polymorphism and phylogenetic relationship in some Meloidogyne spp.: application to the taxonomy of Meloidogyne. AB - Proteins and various isozymes were investigated by direct analysis of single specimens in order to check molecular genetic variability, which is not rare in Meloidogyne species in spite of parthenogenetic reproduction. Variability was found in esterases, ocglycerophosphate, malate dehydrogenases, and some other proteins. Other loci appear monomorphic in the genus (for example, catalase), Distinct pools of genes are in a relative accordance with the common described species. Characteristic electrophoretograms are given for M. arenaria, M. javanica, M. incognita, M. hapla, and M. naasi, and it appears that nonspecific esterases are a useful tool supplementing morphology for specific characterization. Because the biochemical evidence is less subjective than the morphological, we believe it is more reliable. PMID- 19305863 TI - The nematode Heterotylenchus autumnalis and face fly Musca autumnalis: a field study in northern California. AB - Helerolylenchus aulumnalis was found in six northern California counties surveyed, and the incitlence of nematode infection of face flies ranged from 4.7 to 43.8%. Intensive studies at a cattle ranch in Yuba County showed that population densities of the host and nematode infections were highest in flies from cow pats receiving full sun. Average host population density was 105.7 puparia per pat, and nematode infection averaged 38.6%. Pats in partial sun averaged 13.5 puparia and 13,1% nematode infection. No face fly was recovered from shaded pats. When data from pats first exposed during day or night were compared, no significant differences in host population density or nematode infection rates were apparent. Uninfected and superinfected flies were more frequent than predicted by a Poisson distribution.Infected and uninfected female flies of all ages captured on white sticky traps appeared to feed with similar frequency upon a creatny substance which was probably acquired from cattle, However, older infected females fed less on blood and more upon dung than older uninfected females. Percent nematode infection and host population densities were highest in spring and early summer, declined to a midsummer low, and then increased slightly. Both dung-reared flies and captured females showed similar trends in abundance anti infection rates. Regression analysis indicated that H. autumnalis may not be regulating face fly population density. PMID- 19305864 TI - Nematode economic thresholds: derivation, requirements, and theoretical considerations. AB - Determinatitm and use of economic thresholds is considered essential in nematode pest management programs. The economic efficiency of control measures is lnaximized when the difference hetween the crop valne and the cost of pest control is greatest. Since the cost of reducing the nematnde pnpnlation varies with the magnitutle of the reduction attempted, an economic (optimizing) threshold can be determined graphically or mathematically if the nature of the relationships between degree of control and cost, and nematode densities and crop value are known. Economic thresholds then vary according to the nematode control practices used, environmental influences on the nematode damage fnnction, and expected crop yields and values. A prerequisite of the approach is reliability of nematode population assessment techniques. PMID- 19305865 TI - Interaction between Neoaplectana carpocapsae and a granulosis virus of the armyworm Pseudaletia unipuncta. AB - Neoaplectaua carpocapsae developed and reproduced in armyworm hosts infected with a granulosis virus (GV). Macerated tissues of dauer juveniles from GV-infected hosts had sufficient GV to infect 1st and 2nd instar armyworms. Electron microscope examination of dauer juveniles and adult female nematodes confirmed the presence of GV in the lumen of the intestine. No GV was observed in other tissues of the nematode. PMID- 19305866 TI - Meloidogyne grahami n. sp. (meloidogynidae), a root-knot nematode on resistant tobacco in South Carolina. AB - Meloidogyne grahami n. sp. is described and illustrated from specimens on tobacco (Nicotiana tabacum) originally from Florence, South Carolina. Considered for several years to be only a race of M. ineognita, this new species readily attacks NC-95 tobacco, a variety with resistance to the M. incognita group that is common in the major U.S. tobacco-producing areas. M. grahami n. sp. is related most closely to the three subspecies of the M. incognita group but differs from all of them, especially in its distinctive perineal pattern and larger larvae (av. 421 microm, vs. 385 microm or less). Also, the dorsal esophageal gland orifice of females of M. grahami n. sp. is further from the base of the styler (5 microm) than in M. i. incognita and M. i. acrita. Comments are given on the distribution of this new species. PMID- 19305867 TI - Distribution of Nothanguina phyllobia and its potential as a biological control agent for silver-leaf nightshade. AB - The nematode Nothanguina phyllobia Thorne was found within large foliar galls on the perennial weed Solanum elaeagnifolium Cav. in west Texas. A two-year survey of a 6400 sq-km area in west Texas showed extensive distribution of the nematode. No hosts other than S. elaeagnifolium were observed. Densities of juvenile nematodes in the soil were high. N. phyllobia spread rapidly after small numbers of infective juveniles were applied in a foliar spray to an S. elaeagnifolium population. The host plant declined in vigor and frequently died. Artificial inoculation of an S. elaeagnifolium population with large numbers of the nematodes by broadcasting infected plant tissue resulted in high infection incidence. PMID- 19305868 TI - Amino acids and carbohydrates secreted by Meloidogyne incognita. PMID- 19305869 TI - Reaction of banana cultivars to the burrowing nematode (Radopholus similis). PMID- 19305870 TI - Restoration of morphological features of Globodera pallida after storage in formalin. PMID- 19305871 TI - Effects of aldicarb on fusarium wilt-root-knot nematode disease of cotton. PMID- 19305872 TI - Toxicity to Tylenchorhynchus dubius and Hoplolaimus galeatus of newly unwrapped plastic containers. PMID- 19305873 TI - A simple method of processing nematodes for electron microscopy. PMID- 19305874 TI - Effects of oxamyl on the reproduction of Meloidogyne hapla and Heterodera schachtii. PMID- 19305875 TI - [The decline of the medical statute in the beginings of the millenium]. PMID- 19305876 TI - [Yes, we can !]. PMID- 19305877 TI - [Adult cardiac surgery: impact on age group differences in preoperative risk factors and early mortality]. AB - Over the past 20 years, there have been marked increased rates in cardiac surgery among the elderly in Portugal. To evaluate the effects of increasing age on outcome after cardiac surgery, 5652 patients who underwent cardiac surgery from 2003 to 2005 in five portuguese hospitals, were entered into a retrospective study. Patients were placed in five groups according to age: (1) 80 and older, (2) 70-79, (3) 60-69, (4) 50-59 or (5) less than 50. Selected variables included risk factors, cardiac status, preoperative hemodynamics and surgical procedures. In-hospital mortality was collected on all patients. The mean age was 64.7+/-11.6 years-old and younger patients were more often male than older patients (74.4% vs 51.9%). Preoperative COPD and peripheral vascular disease rates increased for those older than 60 years and decreased for those aged 30 to 49 years. Octogenarians had more congestive heart failure (62.5% vs 37.7%), urgent operations (39.2% vs 26.4%), aortic valve surgery (44.5% vs 26,8%) and off-pump CABG (77.8% vs 57.3%) than the younger groups. Hospital mortality was less than 2.0% in patients younger than 69 years-old, 3.5% in septuagenarians and 10,0%in octogenarians. While patients age at operation significantly influenced hospital mortality, this appeared to be a consequence of the increased frequency of risk factors and comorbidities together with decreased physiologic reserve in patients over 80 years of age. PMID- 19305878 TI - [Results of an experience with the Bentall procedure]. AB - OBJECTIVES: The authors report a retrospective study of patients who underwent a Bentall procedure in the Cardio-Thoracic Surgery Department of Hospital Santa Maria, Lisbon. MATERIAL AND METHODS: Data were obtained from 42 patients who underwent the Bentall procedure between 1991 and 2008. Different parameters were compared as well as the short and long term results. RESULTS: The median age was 58,5 years being 88% of patients of male gender. 32 (76,27%) presented with ascending aorta degenerative aneurism, 6 p (14,3%) with type A aortic dissection and 4p (9,5%) with Marfan syndrome. 10p(23.8%) had associated coronary artery disease and there were 6p (14.3%) who underwent emergency surgery. Among the risk factors associated, 24 p (57,1%) had hypertension, 12 p (28,6%) dyslipidemia, 4p (9,5%) had history of smoking, 7 p (16,7%) diabetes, 6 p (14,3%) CPOD, 2 p (4,8%) renal dysfunction and 2 had a recent myocardial infarction. There were 7p (16,7%) with left ventricular dysfunction (EF<40%); 25 p were in NYHA class II-III. The hospitalar mortality was of 2,77% in non-emergent cases, and 33,3% in emergent operations. After 30 days, 92,8% of patients were alive. The 1-year survival was 90,4% and after 5 years of 85,7%. CONCLUSIONS: The Bentall procedure obtained good results in short and long term and, despite the emerging of new and more complex techniques like remodelling and reimplantation, our results support the maintenance of this procedure as the preferential treatment for aortic root and valvular disease. PMID- 19305879 TI - [Right heart thrombus in a patient with pulmonary embolism. Clinical report]. AB - Right heart thrombi are an uncommon finding in pulmonary embolism. The authors report a case of pulmonary embolism in an elderly male, describing its atypical presentation as well as the workup and therapy, followed by a brief thematic review. PMID- 19305880 TI - [Surgical treatment of abdominal aortic aneurysm in patients in hemodialysis program]. AB - INTRODUCTION: The surgical management of abdominal aortic aneurysm in patients with chronic renal insufficiency (CRI) undergoing hemodialysis raises increasing difficulties due to tissue fragility as bleeding and infection susceptibility. PURPOSE: To evaluate the surgical results in patients undergoing hemodialysis submitted to elective repair of the abdominal aortic aneurysm (AAA). PATIENTS AND METHODS: Between March 1982 and January 2008, 942 patients undergoing elective repair of abdominal aortic aneurysm (AAA), being 8 patients (0,8%) in hemodialysis program. The mean age was 61,9+/-3,5 years, six men (75%). The patients were in hemodialysis program in median time of 21,5 months before surgery. Demographics and surgical data were analysed as the perioperative morbidity and mortality rates and causes of late deaths. RESULTS: The mean of operative time was 224,4+/-66 minutes, and blood loss was 812+/-233,12 mL. Major perioperative complications occurred in two patients (25%) and were due to instable angina in one patient and a fatal respiratory infection in another, mortality rate was 12,5%. Median follow up of the survival patients was 25,5 months (ranged 2 to 168 months). Two deaths were noticed during the follow up period, due to myocardial infarction in both cases. CONCLUSIONS: Although the lower number of patients, we conclude that the AAA repair in patients undergoing hemodialysis can be performed with acceptable mortality and morbidity rates and long term survival. PMID- 19305881 TI - [Loco-regional anesthesia in vascular surgery]. AB - Per-operative management of patients with cardiovascular pathology, has been regarded, since ever, as a challenge for anesthesiologists and vascular surgeons. Vascular disease, often diffuse and asymptomatic, has a high prevalence in the community and is associated to an annual mortality rate of 12%. This requires that anesthetic technique should be concentrated in the preservation of myocardial, renal, pulmonary and cerebral functions, during the hemodynamic alterations related to surgery, in order to lower the per-operative complications and to promote faster and safer post-operative recovery. In this paper we describe the experience of the Anesthesiology Department of S. Joatilde;o Hospital, Porto, particulary with the superficial and deep cervical plexus block, in carotid endarterectomy surgery. PMID- 19305882 TI - [Our experience in the surgical management of bull horn vascular injuries]. AB - Vascular trauma caused by bull horn injuries can be regarded as a subgroup of vascular traumatology, due to its particular etiology, mechanism of action, associated injuries and surgical management. In this paper, the authors report their experience in the management of 56 such injuries, for the last 20 years, analysing the annual frequency (more common in summer time), the nature of the injury, the most commonly affected vessels and the surgical reconstruction or revascularization methods employed. The quality of results are discussed according some variable, such as is the case of the precocity of the repair, the degree of contamination, the occurence of post operative infections and the nature of the reconstructive or revascularization method employed. Finnaly, a comparison with similar reports already published in the literature is made. PMID- 19305883 TI - [An exceptional case of renal angiomyolipoma extended to the inferior vena cava. Successfull surgical management]. AB - The clinical case of a 39 years old female is reported, with the diagnosis of tumor of the right kidney extending into the infra-hepatic vena cava, assuming the shape of a floating thrombus. The patient underwent right radical nephrectomy, followed by resection of the intra caval tumor. Both the procedure and post operative course were uneventfull. Histopathological evaluation of the specimens, associated to specific imunohistochemistry studies, confirmed the diagnosis of angiomyolipoma of the kidney. A review of the literature concluded that this is the 27th case published of a kidney angiomyolipoma extending into the inferior vena cava, thus justifying its presentation and divulgation. PMID- 19305884 TI - [Fibromuscular dysplasia of the brachial artery, an unusual cause of upper extremities ischemia. A case report and review of the literature]. AB - Fibromuscular dysplasia (FMD) is a noninflammatory, nonatherosclerotic sistemic disease of unknown etiology, primarily affecting muscular arteries of intermediate size. It has been most commonly observed in the renal, carotid, and intracerebral arteries, although it has been reported in other arterial beds. However, being an uncommon disease in general, the manifestation of FMD in the upper extremities is exceedingly rare. The authors report the case of a 69 years old female admitted with ischemia of the right hand, secondary to fibromuscular dysplasia of the midbrachial artery. The patient presented to vascular surgery clinic with a 4 month history of numbness, pain, and coolness of her right hand, with a small necrotic lesion on her right index finger. Peripheral pulses were barely palpable, and doppler-derived brachial and radial systolic pressures suggested midbrachial artery stenosis. Arteriography showed a normal arch and normal innominate, subclavian, and axillary arteries. The midbrachial artery was markedly abnormal and had alternating areas of stenosis and aneurysm formation - "string-of-beads" appearance. The patient underwent surgical excision of the abnormal right brachial artery, and reconstruction was accomplished with a reversed saphenous vein graft. Distal pulses were restored postoperatively. Pathologic examination confirmed the diagnosis of fibromuscular dysplasia. A review of the literature on the topic was made. PMID- 19305885 TI - [Large myxoma of the left atrium]. PMID- 19305886 TI - Optical differential reflectance spectroscopy of ultrathin epitaxial organic films. AB - This Perspective does not have the ambition to entirely review the subject of optical spectroscopy on thin organic films. What we will try to achieve instead is to give an overview on optical reflectance spectroscopy of highly ordered organic thin films in the thickness range from submonolayers to several monolayers, as a tool to study the absorption behavior of such films. By doing so, we will emphasize the relations between the physical layer structure and the resulting optical properties. More specifically, we intend to show on the basis of particular examples what physical effects can be favorably examined by means of real-time optical spectroscopy, i.e., applied during the actual film growth, especially differential reflectance spectroscopy (DRS). Epitaxial organic films on inorganic substrates (insulators and conductors) will be in focus, and also the perspectives of investigating organic-organic heteroepitaxial layers will be addressed. PMID- 19305887 TI - Charge transfer in Cr adsorption and reaction at the rutile TiO2(110) surface. AB - The rutile TiO2(110) surface has been doped with sub-monolayer metallic Cr, which oxidises and donates charge to specific surface Ti ions. X-Ray and ultra violet photoemission spectroscopy and first principles density functional theory with Hubbard U are used to assign the oxidation states of Cr and surface Ti and we find that Cr2+ forms on bridging oxygen ions and a 5-fold coordinated surface Ti atom is reduced to Ti3+ and the Cr ions readily react with oxygen (to Cr3+), which leads to depletion of surface Ti3+ 3d electrons. PMID- 19305888 TI - Phenol chemisorption onto phthalocyanine thin layers probed by ATR-FTIR difference spectroscopy. AB - Attenuated total reflection Fourier transform infrared (ATR-FTIR) difference spectroscopy has been employed as a powerful method for the comprehension of active layer-analyte interactions, revealing interesting mechanistic aspects concerning the binding of halogen-substituted phenols onto phthalocyanine Langmuir-Schaefer films. PMID- 19305889 TI - Energy transfer processes in electronically coupled porphyrin hetero-dyads connected at the beta position. AB - Energy transfer in a series of hetero-dyads of zinc porphyrin and free-base porphyrin connected at the beta position by pi conjugated bridges has been determined. The dyads have been characterized and compared with the homo dyads, excellent models for the donor and the acceptor porphyrins in the electronically conjugated system. The homo dyads provide reliable parameters for the determination of the energy transfer rate calculated according to the Forster theory. This model was inadequate to account for the experimental findings and an electron exchange mechanism was shown to contribute. A favorable coplanar arrangement of the bridge and the tetrapyrroles facilitates the energy transfer process, which displays a very low distance dependence and an efficiency >98%. PMID- 19305890 TI - Au nanoparticles on a templating TiO(x)/Pt(111) ultrathin polar film: a photoemission and photoelectron diffraction study. AB - We present an in-depth investigation of Au nanoparticles self-assembled on a zigzag-like TiO(x)/Pt(111) ultrathin polar film, whose structure is known in great detail. The peculiar pattern of defects (picoholes) templates a linear array of size-selected (ca. 1 nm) Au nanoparticles without disruption of the titania layer, as observed by scanning tunneling microscopy. Their structure and electronic properties have been investigated by several large-area spectroscopic tools, i.e. high-resolution core and valence level photoemission and angle scanned and energy-scanned photoelectron diffraction. The comparison between experimental data and density functional theoretical calculations indicates that the Au atoms landing on the oxide film are rather mobile, and that the picoholes can act as effective trapping and nucleation centers for the growth of the Au nanoparticles. All the experimental results are in concord in indicating that the Au NPs are flat islands with a maximum thickness of 2-3 layers exposing the (111) surface. PMID- 19305891 TI - Hierarchical pattern of microfibrils in a 3D fluorapatite-gelatine nanocomposite: simulation of a bio-related structure building process. AB - The shape development of a biomimetic fluorapatite-gelatine nanocomposite on the mum scale is characterised by a fractal mechanism with the origin being intrinsically coded in a (central) elongated hexagonal-prismatic seed. The 3D superstructure of the seed is distinctively overlaid by a pattern consisting of gelatine microfibrils. The orientation of the microfibrils is assumed to be controlled by an intrinsic electrical field generated by the nanocomposite during development and growth of the seed. In order to confirm this assumption and to get more detailed information on orientational relations of the complex nanocomposite we simulated the pattern formation process up to the microm scale. The results from experimental studies and simulation results on an atomistic level support a model scenario wherein the elementary building blocks for the aggregation are represented by elongated hexagonal-prismatic objects (A-units), with the embedded collagen triple-helices in their centers. The interactions of the A-units are consequently modelled by three contributions: the crystal energy part (originating from the pair-wise interactions of the "apatite shells" of the prismatic units), the electrostatic interaction (originating from the unit charges located at the ends of the collagen triple helices), and the interaction energy of the A-units mediated by the solvent. The next level of complexity is related to the fact that micro fibrils were found in the fluorapatite-gelatine nanocomposites. They consist of bundles of triple helical protein molecules, which are embedded within the 3D-hexagonal prismatic arrangement of the A-units. In our approach we consider the microfibrils as chains of flexible dipoles with effective dipole moments. The crystal growth processes is modelled as an energetically controlled stepwise association of elementary building blocks of different kind on a 3D-grid. The remarkable and excellent qualitative agreement between the simulated fibril patterns and the observations made by SEM and TEM support the concept of an intrinsic electric field driven morphogenesis of the fluorapatite-gelatine nanocomposite. The simulated fibril pattern also bears the chance to make fresh attempts in order to find explanations for experimental observations which are not understood up to now. PMID- 19305892 TI - Improvement of the capacitive performances for Co-Al layered double hydroxide by adding hexacyanoferrate into the electrolyte. AB - This paper reports on the improvement on the capacitive properties of Co-Al layered double hydroxide (Co-Al LDH) by adding hexacyanoferrate(II) and (III) solely or jointly into 1 M KOH aqueous solution. Owing to the high reversibility, the Fe(CN)(6)(3-)/Fe(CN)(6)(4-) ion pair acts as an electron relay at the electrode/electrolyte interface during charge and discharge by coupling in the redox transition of Co(II)/Co(III) in the Co-Al LDH electrode. Electrochemical impedance spectra and Tafel curves provide direct evidences with decreased charge transfer resistance and increased exchange current density in the alkaline solution containing hexacyanoferrate ions, respectively. PMID- 19305893 TI - Redox switching and oxygen evolution electrocatalysis in polymeric iron oxyhydroxide films. AB - Outstanding issues regarding the redox switching characteristics and the oxygen evolution reaction (OER) electrocatalytic behaviour of multicycled iron oxyhydroxide films in aqueous alkaline solution have been examined. Charge percolation through the hydrous layer has been quantified, using cyclic voltammetry, in terms of a charge transport diffusion coefficient D(CT) which admits a value of ca. 3 x 10(-10) cm2 s(-1). Steady-state Tafel plot analysis and electrochemical impedance spectroscopy have been used to elucidate the kinetics and mechanism of oxygen evolution. Tafel slope values of ca. 60 mV dec(-1) and ca. 120 mV dec(-1) are found at low and high overpotentials respectively, whereas the reaction order with respect to hydroxide ion activity changes from ca. 3/2 to ca. 1 as the potential is increased. These observations are rationalised in terms of a kinetic scheme involving Temkin adsorption and the rate determining formation of a physisorbed hydrogen peroxide intermediate on the oxide surface. The dual Tafel slope behaviour is ascribed to the potential dependence of the surface coverage of adsorbed intermediates. PMID- 19305894 TI - Chromocene in porous polystyrene: an example of organometallic chemistry in confined spaces. AB - In this work, we present an innovative approach to investigate the structure and the reactivity of a molecularly dispersed organometallic compound. The poly(4 ethylstyrene-co-divinylbenzene) microporous system (PS) is used as "solid solvent" able to molecularly disperse CrCp2, allowing: (i) its full characterization by means of spectroscopic techniques; (ii) the pressure and temperature dependent study of its interaction towards simple molecules like CO freely diffusing through the pores; (iii) the accurate determination of the reaction enthalpies by both direct microcalorimetric measurements and by an indirect spectroscopic approach. The experimental results are compared with quantum-mechanical calculations adopting the DFT approximation with two different functionals (namely BP86 and B3-LYP), showing the limitations and the potentialities of DFT methods in predicting the properties of open shell systems. It is concluded that modern DFT methods are able to give a coherent view of the vibrational properties of the CrCp2 molecule (and of the complex formed upon CO adsorption) that well match the experimental results, while the energetic predictions should be taken with care as they are significantly dependent on the functionals used. PMID- 19305895 TI - Combination of (100), (110) and (111) facets in MgO crystals shapes from dry to wet environment. AB - At the onset of dissolution in water, cubic MgO smoke crystals present (110) cuts of the edges of the cubes. Next, (111) facets progressively dominate the shape of the crystallites, which finally transform into truncated octahedra. The morphology of the crystallites that are derived from surface energies computed within the density functional theory (DFT), only involve (100) and (111) facets. We explain the unexpected (110) cuts via a "constrained" Wulff equilibrium shape that arises from a slower kinetics of formation of (111) facets than (110) ones. Experiment and theory fully agree on the hierarchy of hydroxylated surface energies: Gamma(111) < Gamma(100) < Gamma(110), both supporting the partial dissociation of water on MgO(100). Finally, from low to high P(H2O) (high to low T), DFT-based calculations predict a switch from Wulff shapes involving dry (100) facets, in which the (100)/(111) area ratio decreases upon increasing P(H2O), to shapes involving hydroxylated (100) surfaces, in which the above ratio increases with P(H2O). PMID- 19305896 TI - Multiphoton dissociation dynamics of CH3Br. AB - The state-resolved photodissociation dynamics of CH3Br+ cations are studied using a combination of slice imaging and velocity mapping. Parent ions are prepared by (2 + 1) two-photon resonant three-photon ionization of CH3Br through the 5s Rydberg state in the energy region 29,069.8-29,976.0 cm(-1). Analysis of the photoelectron spectra reveals that CH3Br+ ions are produced in the spin-orbit ground electronic state with a wide distribution of vibrational excitation. The photodissociation of CH3Br+ produces CH3+ (X) + Br(2P(1/2))/Br(2P(3/2)) whose product state distribution shows substantial excitation in the umbrella mode of the CH3+ photofragment, especially when concurrently producing the Br(2P(1/2)) channel. The shapes of the photofragment angular distributions clearly show the different dissociation mechanisms in the Br(2P(1/2)) and Br(2P(3/2)) channels. The dissociation dynamics of the 5s Rydberg state of CH3Br are also presented and analyzed. PMID- 19305897 TI - MRCI investigation of Ni2O2 and Ni2O2+. AB - The geometrical and electronic structures of rhombic Ni2O2 and Ni2O2+ are investigated by multireference configuration interaction (MRCI) calculations using natural atomic orbital basis sets. The ground term of Ni2O2 is a 1A(g) term with a Ni-O distance of 178.9 pm and a Ni-O-Ni angle of 79.6 degrees. At energies below 1 eV there are at least 21 further terms of singlet, triplet, and quintet multiplicity. The lowest-lying term of Ni2O2+ is a 2B1 term with a distorted C2v symmetric structure with two non-equivalent Ni centres and Ni-O distances of 170.6 and 182.6 pm. A 4B2 term is found at an energy higher by only 0.02 eV. At energies below 0.6 eV there are at least 6 further terms of doublet and quartet multiplicity. The lowest-lying sextet terms appear at about 0.6 eV. The calculated Ni-O-Ni angle of Ni2O2 is in very close agreement with the value derived from IR spectra. The fact of a doublet ground state of Ni2O2+ does not support the assumption that spin restrictions are the reason for the absence of any observable reaction of Ni2O2+ with H2 and CH4. PMID- 19305898 TI - Mass analysed threshold ionisation spectroscopy of flexible 2-para fluorophenylethanol conformers with and without an intramolecular OH...pi bond. AB - The cationic state of the prototype flexible molecule 2-para-fluorophenylethanol has been investigated by combination of mass-analysed threshold-ionisation (MATI) spectroscopy and quantum chemistry ab initio density functional theory (DFT) calculations employing two different functionals: the B3LYP functional and the new hybrid functional M05. The MATI spectra measured via vibronic bands in the S1 intermediate state belonging to the most stable gauche conformer stabilised by an intramolecular OH...pi hydrogen bond are structureless, while the spectra recorded via bands of the anti conformer feature well-resolved peaks. This result is in a good accord with our theoretical predictions showing that upon ionisation, the anti conformer retains its structure, while the lowest-energy gauche conformer undergoes a significant structural change resulting in a break of the OH...pi bond. This and the good agreement between the measured band positions and the theoretically predicted frequencies for the cationic anti conformer confirm the conformational assignment. The result for the cation provides clear evidence for the existence of a nonclassical intramolecular OH...pi hydrogen bond if the electron density in the aromatic ring is sufficiently high as is the case only for the neutral molecule. PMID- 19305899 TI - A surprisingly complex aqueous chemistry of the simplest amino acid. A pulse radiolysis and theoretical study on H/D kinetic isotope effects in the reaction of glycine anions with hydroxyl radicals. AB - A pulse radiolysis study was carried out of the reaction rate constants and kinetic isotope effects of hydroxyl-radical-induced H/D abstraction from the most simple alpha-amino acid glycine in its anionic form in water. The rate constants and yields of three predominantly formed radical products, glycyl (NH2-*CH-CO2-), aminomethyl (NH2-*CH2), and aminyl (*NH-CH2-CO2-) radicals, as well as of their partially or fully deuterated analogs, were found to be of comparable magnitude. The primary, secondary, and primary/secondary H/D kinetic isotope effects on the rate constants were determined with respect to each of the three radicals. The unusual variety of products for such an elementary reaction between two small and simple species indicates a complex mechanism with several reactions taking place simultaneously. Thus, a theoretical modeling of the reaction mechanism and kinetics in the gas- and aqueous phase was performed by using the unrestricted density functional theory with the BB1K functional (employing the polarizable continuum model for the aqueous phase), unrestricted coupled cluster UCCSD(T) method, and improved canonical variational theory. Several hydrogen-bonded prereaction complexes and transition states were detected. In particular, the calculations pointed to a significant mechanistic role of the three-electron two orbital (sigma/sigma* N therefore O) hemibonded prereaction complexes in the aqueous phase. A good agreement with the experimental rate constants and kinetic isotope effects was achieved by downshifting the calculated reaction barriers by 3 kcal mol(-1) and damping the NH(D) stretching frequency by a factor of 0.86. PMID- 19305900 TI - Effect of adsorbate mass on an Eley-Rideal reaction. Reactive scattering of Cs+ from noble gases and N2 adsorbed on Ru(0001) surfaces at hyperthermal energy. AB - The effect of the adsorbate mass on an Eley-Rideal (ER) reaction is examined for the reactive ion scattering (RIS) of Cs+ from physisorbed adsorbates (Ar, Kr, Xe and N2) on a Ru(0001) surface at hyperthermal energy (10-60 eV). The measurement of the RIS cross-section, or the efficiency for ER-type abstraction of surface adsorbates, reveals an inverse proportionality between the square root of the cross-section and the mass of the noble gases. This relationship is explained by the inertia effect which can determine the probability of successful abstraction events of the adsorbates. The result confirms a theoretical prediction that the inertia effect is important for the dynamics of an ER reaction in the hyperthermal energy regime. PMID- 19305901 TI - Electrochemical behaviour of hen-egg-white lysozyme at the polarised water/1,2 dichloroethane interface. AB - The electrochemical behaviour of hen-egg-white lysozyme (HEWL) was studied at the polarized water/1,2-dichloroethane interface. The voltammetric ion-transfer response was found to be dependent on the pH and ionic strength of the aqueous phase solution and also on the nature of the organic phase electrolyte anion. The current-pH behaviour of HEWL was dominated by the charge of the biomolecule at each pH, as indicated by the close relationship between the experimental peak currents and the theoretical curve for HEWL based on its known acid-base chemistry. Three organic electrolyte anions of differing hydrophobicities were investigated (TFPB-, TPBCl- and TPB-) and it was found that the ion transfer voltammetric peaks occurred at successively higher potentials in the order of increasing hydrophobicity, Deltaphi(TPB) < Deltaphi(TPBCl) < Deltaphi(TPBF). The voltammetric response was time dependent during multi-cyclic voltammetry experiments, with the formation of a white film of precipitate at the interface. A pre-peak consistent with adsorption of the HEWL ion transfer product at the liquid/liquid interface was also observed. The results suggest that an adsorption or re-arrangement of HEWL molecules with time at the interface is taking place. A mechanism for the response on application of a triangular potential waveform with cyclic voltammetry is proposed based on an i-C-i mechanism. Our results indicate that HEWL is electroactive at the polarized liquid/liquid interface and that such electrochemical methods may provide an approach to the label-free detection and characterization of protein molecules. PMID- 19305902 TI - Hydrogen bonding in 1,2-diazine-chloranilic acid (2 : 1) studied by a 14N nuclear quadrupole coupling tensor and multi-temperature X-ray diffraction. AB - Protons involved in the H-bond system in 1,2-diazine-chloranilic acid (2 : 1) are assumed to be in jumping motion in the double-minimum potential corresponding to the two extreme electronic states of O-H...N and O-...H-N+. 14N nuclear quadrupole coupling constants were determined by 1H-14N nuclear quadrupole double resonance. Assuming that the observed coupling constants are result of a fast exchange of the two extreme electronic states, the coupling constants for each state were estimated by use of the equilibrium populations of the two extreme states determined from multi-temperature X-ray single-crystal diffraction. It was suggested that not only the population but also the electron distribution of the extreme electronic states itself changes with temperature. PMID- 19305903 TI - Lifetime and diffusion coefficient of active oxygen species generated in TiO2 sol solutions. AB - Active oxygen species generated by photoexcitation of TiO2 sol solutions were directly observed by the single-shot near-field heterodyne transient grating method. Transient responses were compared in the presence and absence of various kinds of scavengers such as sodium azide, dimethyl sulfoxide, and superoxide dismutase, and three components of the transient responses were assigned to hydroxyl radicals, hydrogen peroxide, and superoxide radicals. The diffusion coefficients of each active oxygen species were obtained by the analysis of the transient responses, and it was revealed that they were 1-3 orders smaller than those for molecules with a similar size. It was proposed that this is because the active oxygen species are under equilibrium between free radicals and adsorbed species on a TiO2 surface. Furthermore, it was suggested that the adsorption equilibrium for each species was varied depending on the pH of the solutions. PMID- 19305904 TI - Analytic ab initio calculations of coherent anti-Stokes Raman scattering (CARS). AB - We present a theory for the analytic calculation of frequency-dependent polarizability gradients, and apply the methodology to the calculation of coherent anti-Stokes Raman scattering (CARS). The formalism used is based on an open-ended theory for the calculation of frequency-dependent molecular response properties of arbitrary order, also including contributions from perturbation dependent basis sets. An important feature of our approach is the close connection between the formalism--which is fully matrix-based in an atomic orbital basis--and the implementation, allowing for the rapid implementation of higher-order molecular properties. Care is taken to allow the formalism to be utilized with linearly-scaling Hartree-Fock and density-functional theory codes. By avoiding the evaluation of responses due to geometry distortions, only 9 response equations need to be solved for the calculation of the CARS intensities, independent of the size of the molecular system. The theory is illustrated by calculations on a set of polyaromatic hydrocarbons using a DFT/B3LYP force field and Hartree-Fock polarizability gradients. Good agreement with the experimental CARS spectra of these compounds is obtained. PMID- 19305905 TI - Effects of composition on structure and activity of PtRu/C catalysts. AB - A series of carbon supported PtRu bimetallic catalysts with varying Pt:Ru ratio were prepared and characterised using ex situ and in situ XRD, in situ EXAFS at 0 V vs. RHE, ex situ XPS and monolayer CO stripping voltammetry. Although the catalysts were found to be well mixed/alloyed, with no evidence of unalloyed Ru (oxides) present, the surfaces of the electrocatalyst nanoparticles were found to be enriched with Pt compared to the nominal bulk composition. The methanol oxidation activities of the catalysts were determined in 1.0 mol dm(-3) H2SO4. In agreement with published studies of polycrystalline bulk PtRu alloys the catalyst with a 0.6 surface fraction of Pt was found to give the best methanol oxidation activity at 30 degrees C. However, at 80 degrees C a greater surface fraction of Ru could be tolerated, with some activity at low current densities found for a Pt surface fraction as low as 0.2. The results support the conclusion that a limited amount of methanol dehydrogenation occurs at Ru sites or Ru dominated surface ensembles at 80 degrees C. PMID- 19305906 TI - Mesoscopic and microstructural characterization of liposomes formed by the lipooligosaccharide from Salmonella minnesota strain 595 (Re mutant). AB - Large unilamellar liposomes formed by the Re LPS (from here below named LOS) from Salmonella enterica serotype minnesota strain 595 (Re mutant) have been prepared by the extrusion techniques. The physico-chemical investigation of these systems has been performed by a combined experimental strategy, which has allowed a characterization at different observation scales, from the morphological to the micro-structural level. Particularly, dynamic light scattering (DLS) measurements have revealed the formation of liposomes with a narrow dimension distribution; small-angle neutron scattering (SANS) measurements have led to an estimation of the thickness of the LOS bilayer, which is consistent with an extended conformation of the acyl chains; electron paramagnetic resonance (EPR) measurements have allowed to investigate the bilayer fluidity, as determined by LOS acyl chain conformation and packing; Pulsed-field-gradient stimulated echo nuclear magnetic resonance (PGSTE-NMR) measurements have shown that water permeability through the LOS bilayer is low. The effect of temperature has been considered. Liposome dimension does not change; however, centered at 30-35 degrees C, a progressive transition of the acyl chain self-organization from a gel to a liquid crystalline phase is detected. Above this temperature, the water permeability through the bilayer increases. The effect of introducing the zwitterionic phospholipid dilauroyl phosphatidylcholine (DLPC) in the liposome formulation have been also analysed. A comparative analysis of the results points out a clear relation between the microstructure of LOS molecules in the bilayer and liposome functional properties. PMID- 19305907 TI - Temperature dependence of the rate coefficient for the alpha-pinene reaction with ozone in the range between 243 K and 303 K. AB - The absolute rate coefficient for the reaction of alpha-pinene with ozone was determined in the temperature range between 243 K and 303 K at atmospheric pressure. In total, 30 experiments were performed in the large (85 m3) temperature-controlled simulation chamber AIDA, where the concentrations of the reactants ozone and alpha-pinene were measured directly. An Arrhenius expression for the alpha-pinene + ozone reaction was derived with a pre-exponential factor of (1.4 +/- 0.4) x 10(-15) cm3 s(-1) and a temperature coefficient of (833 +/- 86) K. This rate coefficient is in good agreement (-5%) with the current IUPAC (IUPAC 2007) recommendation at 298 K. The IUPAC recommendation is significantly larger (+27%), around 243 K where the recommended values were extrapolated from higher temperatures. This finding is relevant for tropical regions where strong updrafts can rapidly transport reactive hydrocarbons like alpha-pinene from the boundary layer into the cold regions of the free troposphere. PMID- 19305908 TI - A density functional study on cationic Au(n)Cu(m)+ clusters and their monocarbonyls. AB - Small cationic Au(n)Cu(m)+ (n + m < or = 6) clusters and their monocarbonyls Au(n)Cu(m)CO+ have been studied by first-principles calculations. The trend for small Au clusters to form planar structures is weaker when some Au atoms are substituted by Cu atoms. A significant odd-even oscillation of the electron affinity of the cationic clusters with the number of their Au or Cu atoms is observed. CO prefers binding to Cu, which can be understood by the frontier molecular orbital theory. Its binding energy on Au(n)Cu(m)+ generally decreases with the increase of the Cu content in the cluster, which is highly related to the electron transfer between CO and the cluster. Our calculation suggests that reactive collision between CO and Au3Cu+ may lead to the dissociation of the cluster with an Au atom loss. PMID- 19305909 TI - Are the silyl group hydrogens in peri-substituted-9-silyltriptycenes engaged in blue-shifting hydrogen bonds? AB - In three 9-silyltriptycenes bearing chlorine, bromine and the methyl group in one of the peri positions, the silyl group suffers extreme hindrance of its reorientational motion. Owing to this, separate signals of the individual silyl group protons could be observed in NMR spectra at relatively high temperatures. For each compound, the measured values of J couplings of these protons to the 29Si nucleus are strongly diversified, especially when a halogen atom occurs in the peri position. These differences can be qualitatively interpreted in terms of electron density transfers from the peri substituent, causing the individual Si-H bonds to differentiate. The stereoelectronic picture revealed by natural bond orbital analysis fits the pattern normally interpreted as that of the blue shifting hydrogen bond. However, an essential similarity of the stereoelectronic effects for both the halogen and methyl substituents raises a serious question about the applicability of that concept to the present case. PMID- 19305910 TI - Amphiphilic nanoassemblies for the detection of peptides and proteins using fluorescence and mass spectrometry. AB - Amphiphilic nanostructures provide unique environments for molecules that are incompatible with the solvent to be sequestered within their interior. These internal environments provide opportunities for concentrating an analyte or transducer molecule for detection, and the functional groups within the amphiphiles provide an opportunity for incorporating specificity or selectivity toward analytes. In this review, we discuss ways in which amphiphilic assemblies can be used to detect peptides and proteins with a particular emphasis on facially amphiphilic polymers and dendrimers. PMID- 19305911 TI - Peptide microarrays for carbohydrate recognition. AB - An application of high density random sequence peptide microarrays for rapid and reliable identification of artificial carbohydrate receptors is reported. PMID- 19305912 TI - Self-assembly of supramolecular aptamer structures for optical or electrochemical sensing. AB - The self-assembly of labeled aptamer sub-units in the presence of their substrates provides a method for the optical (fluorescence) or electrochemical detection of the substrate. One of the sub-units is linked to CdSe/ZnS quantum dots (QDs), and the self-assembly of the dye-functionalized second sub-unit with the modified QDs, in the presence of cocaine, stimulates fluorescence resonance energy transfer (FRET). This enables the detection of cocaine with a detection limit corresponding to 1 x 10(-6) M. Alternatively, the aptamer fragments are modified with pyrene units. The formation of a supramolecular aptamer-substrate complex allosterically stabilizes the formation of excimer supramolecular structure, and its characteristic emission is observed. In addition, the thiolated aptamer sub-unit is assembled on an Au electrode. The Methylene Blue labeled sub-unit binds to the surface-confined fragment in the presence of cocaine. The amperometric response of the system allows the detection of cocaine with a detection limit of 1 x 10(-5) M. The approach is generic and can be applied to other substrates, e.g. adenosine triphosphate. PMID- 19305913 TI - The preferential electrocatalytic behaviour of graphite and multiwalled carbon nanotubes on enediol groups and their analytical implications in real domains. AB - Carbon nanotubes (CNTs) possess preferential 'electrocatalytic' properties that affect the oxidation of enediol groups, establishing a relationship between electrocatalysis and chemical structure. Since this chemical structure occurs in analytes involved in high impact areas such as the pharmaceutical, cosmetic, and food safety industries, this preferential electrochemical behaviour was demonstrated using both standard and selected real-world samples. The oxygen containing species present on the surface of CNTs and generated during acid treatment were responsible for an enhanced electron transfer reaction for these structures using a proton-assisted electron transfer mechanism, thus confirming their crucial role during the surface preparation process of electrocatalysis. The analytical benefits were that the inherent selectivity and sensitivity from these nanomaterials could be exploited for the direct detection of analytes in complex matrices, revealing their crucial role in the simplification of analytical processes. PMID- 19305914 TI - Predicting sorption of pharmaceuticals and personal care products onto soil and digested sludge using artificial neural networks. AB - A comprehensive analytical investigation of the sorption behaviour of a large selection of over-the-counter, prescribed pharmaceuticals and illicit drugs to agricultural soils and freeze-dried digested sludges is presented. Batch sorption experiments were carried out to identify which compounds could potentially concentrate in soils as a result of biosolid enrichment. Analysis of aqueous samples was carried out directly using liquid chromatography-tandem mass spectrometry (LC-MS/MS). For solids analysis, combined pressurised liquid extraction and solid phase extraction methods were used prior to LC-MS/MS. Solid water distribution coefficients (K(d)) were calculated based on slopes of sorption isotherms over a defined concentration range. Molecular descriptors such as log P, pK(a), molar refractivity, aromatic ratio, hydrophilic factor and topological surface area were collected for all solutes and, along with generated K(d) data, were incorporated as a training set within a developed artificial neural network to predict K(d) for all solutes within both sample types. Therefore, this work represents a novel approach using combined and cross validated analytical and computational techniques to confidently study sorption modes within the environment. The logarithm plots of predicted versus experimentally determined K(d) are presented which showed excellent correlation (R(2) > 0.88), highlighting that artificial neural networks could be used as a predictive tool for this application. To evaluate the developed model, it was used to predict K(d) for meclofenamic acid, mefenamic acid, ibuprofen and furosemide and subsequently compared to experimentally determined values in soil. Ratios of experimental/predicted K(d) values were found to be 1.00, 1.00, 1.75 and 1.65, respectively. PMID- 19305915 TI - Data handling of complex GC-MS chromatograms: characterization of n-alkane distribution as chemical marker in organic input source identification. AB - The paper describes a signal method for processing GC-MS signals to extract usable information hidden in the chromatogram thus reducing the labour and time required to handle the data and increasing the quality and objectivity of the results. The method is focused on two relevant parameters for identification and characterization of the n-alkane series present in complex samples (in particular the C(14)-C(33) terms): the number of n-alkanes, n(max), and the Carbon Preference Index (CPI) describing the odd/even carbon-number predominance. This is a key diagnostic parameter to determine the biogenic and anthropogenic nature of n-alkane sources, useful as chemical markers in source identification and differentiation. The method is a further extension of the approach based on the AutoCovariance Function (ACVF(tot)): new mathematical equations have been derived and a new computation algorithm implemented to extract information on the n alkane series--n(max) and CPI--directly from the EACVF(tot) computed on the acquired chromatographic signal. The procedure was validated on simulated chromatograms where the distribution of the terms of the series describing experimental GC signals was known: the obtained results prove that the parameters n(max) and CPI of the homologous series can be estimated with good accuracy and precision. The method applicability was tested on experimental chromatograms of real samples: gasoils and plant extracts were studied to identify n-alkane distribution patterns characteristic of petrogenic and natural samples. PMID- 19305916 TI - Tailored-waveform collisional activation of peptide ion electron transfer survivor ions in cation transmission mode ion/ion reaction experiments. AB - Broadband resonance excitation via a tailored waveform in a high pressure collision cell (Q2) on a hybrid quadrupole/time-of-flight (QqTOF) tandem mass spectrometer has been implemented for cation transmission mode electron transfer ion/ion reactions of tryptic polypeptides. The frequency components in the broadband waveform were defined to excite the first generation intact electron transfer products for relatively large tryptic peptides. The optimum amplitude of the arbitrary waveform applied has been determined empirically to be 3.0 V(p-p), which is effective for relatively high mass-to-charge (m/z) ratio precursor ions with little elimination of sequence information for low m/z ions. The application of broadband activation during the transmission mode ion/ion reaction obviates frequency and amplitude tuning normally associated with ion trap collision induced dissociation (CID). This approach has been demonstrated with triply and doubly charged tryptic peptides with and without post-translational modifications. Enhanced structural information was achieved by production of a larger number of informative c- and z-type fragments using the tailored waveform on unmodified and modified (phosphorylated and glycosylated) peptides when the first generation intact electron transfer products fell into the defined frequency range. This approach can be applied to a wide range of tryptic peptide ions, making it attractive as a rapid and general approach for ETD LC-MS/MS of tryptic peptides in a QqTOF instrument. PMID- 19305917 TI - Automated molecular formula determination by tandem mass spectrometry (MS/MS). AB - Automated software was developed to analyze the molecular formula of organic molecules and peptides based on high-resolution MS/MS spectroscopic data. The software was validated with 96 compounds including a few small peptides in the mass range of 138-1569 Da containing the elements carbon, hydrogen, nitrogen and oxygen. A Micromass Waters Q-TOF Ultima Global mass spectrometer was used to measure the molecular masses of precursor and fragment ions. Our software assigned correct molecular formulas for 91 compounds, incorrect molecular formulas for 3 compounds, and no molecular formula for 2 compounds. The obtained 95% success rate indicates high reliability of the software. The mass accuracy of the precursor ion and the fragment ions, which is critical for the success of the analysis, was high, i.e. the accuracy and the precision of 850 data were 0.0012 Da and 0.0016 Da, respectively. For the precursor and fragment ions below 500 Da, 60% and 90% of the data showed accuracy within < or = 0.001 Da and < or = 0.002 Da, respectively. The precursor and fragment ions above 500 Da showed slightly lower accuracy, i.e. 40% and 70% of them showed accuracy within < or = 0.001 Da and < or = 0.002 Da, respectively. The molecular formulas of the precursor and the fragments were further used to analyze possible mass spectrometric fragmentation pathways, which would be a powerful tool in structural analysis and identification of small molecules. The method is valuable in the rapid screening and identification of small molecules such as the dereplication of natural products, characterization of drug metabolites, and identification of small peptide fragments in proteomics. The analysis was also extended to compounds that contain a chlorine or bromine atom. PMID- 19305918 TI - Detection of drugs and their metabolites in dusted latent fingermarks by mass spectrometry. AB - A hydrophobic silica dusting agent containing carbon black has been used with latent finger marks to demonstrate that the agent can act as an enhancing matrix to generate a simple method for detecting a range of drugs using surface assisted laser desorption/ionisation time-of-flight mass spectrometry (SALDI-TOF-MS) in positive ion reflectron mode. The dusting agent produces developed marks for locating/visualising the prints whilst also acting as a SALDI-TOF-MS enhancer that is equivalent to the standard matrix enhancer 2,5-dihydroxybenzoic acid. This method has been applied to the analysis of latent fingermarks for contact residues on fingers, and for detection of illicit drugs for both parent drugs and their metabolites. Analysis was performed by direct MS analysis of the pre-dusted fingermarks on the surface of a target plate and, following lifting using commercial tape, MS analysis of the lifted marks. When 19 commercial powders were used only three produced MS spectra but with intensities less than those produced with the new powder. The presence of the parent drug and its metabolites was confirmed using SALDI-TOF-MS-MS following high energy collision induced dissociation when characteristic and unique fragmentation patterns were observed in each case. The distribution of these compounds on fingermarks was subsequently demonstrated using commercially available imaging software. PMID- 19305919 TI - Improved diagnostic accuracy of malignant neck lumps by a simple BMVC staining assay. AB - A handheld device based on fluorescence of 3,6-bis(1-methyl-4 vinylpyridinium)carbazole diiodide (BMVC) staining was established for the rapid, point-of-care screening of cancer cells (see Chang and co-workers, Analyst, 2007, 132, 745). Offering instant screening of cancer at low cost, here we apply this simple assay in clinical tests on fine needle aspirates of neck masses from 114 outpatients (115 specimens). The diagnostic accuracy of this simple method alone is ca. 80% (80/99). The combination of the BMVC test and the fine needle aspiration (FNA) cytology reduced the non-diagnosis from 17 cases in FNA cytology to 6 cases in the combined method. Moreover, an algorithm is proposed to improve the diagnostic accuracy of malignant neck lumps up to nearly 100%. PMID- 19305920 TI - Data processing technique in gas chromatography/time-of-flight mass spectrometry. AB - A complete mass spectrum was measured at an acquisition rate of 1 kHz using a femtosecond-multiphoton-ionization time-of-flight mass spectrometer for analyte eluting from a gas chromatograph. A contour-map display, i.e. a two-dimensional plot, of the data was first constructed for a simultaneous overview of the mass spectra and the mass chromatograms. The noise component in the signal was then reduced by digital filtering and reduced further by smoothing the data. The detection limit (S/N = 3) was improved from 240 to 76 fg for 2,3,4,6,7 pentachlorodibenzofuran, with no appreciable loss of resolution in the chromatogram. The achieved value was lower than the value (<100 fg) required by Japanese Industrial Standards for dioxin analysis. A new parameter, referred to as a 'reliability factor', was defined in this study to calculate the similarity between the intensity distribution of the isotope peaks observed in the mass spectrum and that calculated from the natural abundance ratio of the isotopes. This parameter was useful for evaluation of the purity of the chromatographic peak. PMID- 19305921 TI - Peptide-imprinted polymer microspheres prepared by precipitation polymerization using a single bi-functional monomer. AB - A single bi-functional monomer, N,O-bismethacryloyl ethanolamine (NOBE), was used in precipitation polymerization system to synthesize molecularly imprinted polymer (MIP) microspheres. Highly specific binding sites were obtained for N terminal protected neuropeptides, Boc-Leu-enkephalin and Pyr-Leu-enkephalin. The use of NOBE allowed binding sites to be formed in polymer microspheres that are able to recognize target peptides through the consensus C-terminal sequence. The interesting molecular binding results suggest a new approach for peptide analysis combining in situ chemical modification with MIP recognition under non-aqueous conditions. PMID- 19305922 TI - Quadruple-allele chemiluminometric assay for simultaneous genotyping of two single-nucleotide polymorphisms. AB - We developed a rapid, simple, cost-effective and high sample-throughput method for the simultaneous detection of four alleles in single-nucleotide polymorphisms (SNPs). The method was applied to the simultaneous genotyping of two common SNPs within the TLR4 gene, the A896G and C1196T polymorphisms. The method consists of a single PCR of the region spanning the A896G and C1196T polymorphic sites, followed by a quadruple primer extension (PEXT) reaction in a single tube. A biotinylated nucleotide is incorporated in the extended primer. All four products are captured in streptavidin (SA)-coated microtiter wells and detected with a combination of four reporters, the photoprotein aequorin (AEQ) and the enzymes alkaline phosphatase (ALP), beta-galactosidase (GAL) and horseradish peroxidase (HRP). For each SNP, 46 individuals were genotyped. The accuracy of this method was confirmed by sequencing. The proposed quadruple-allele chemiluminometric assay provides an accurate, simple, rapid, reproducible and cost-effective method for high sample-throughput genotyping of single-nucleotide polymorphisms. PMID- 19305923 TI - Fabrication of an electrically heated indium-tin-oxide electrode for electrochemiluminescent detection system. AB - An electrochemiluminescence (ECL) detection system equipped with an electrically heated indium-tin-oxide (ITO) electrode was constructed. The heated ITO electrode (HITOE) coupled the advantages of a heated electrode with the optical transparency property of ITO glass. Compared with the conventional heated electrode, the HITOE is integrated into the ECL cell, which could save the expensive luminescent reagent. The temperature of the electrode (T(e)) could be accurately controlled by electrically heating. The Ru(bpy)(3)(2+)/TPrA ECL system and the colchicine/Ru(bpy)(3)(2+) ECL system were used to evaluate the developed ECL detection system. The higher sensitivity and lower RSD of ECL detection for tri-n-propylamine (TPrA) and colchicine were gained by moderately heating the electrode. The proposed method has been successfully utilized to the assay of colchicine in a spiked human serum sample. The results indicated that the developed ECL detection system could provide high sensitivity and excellent reproducibility for analytical practice. PMID- 19305924 TI - Label-free detection of B. anthracis spores using a surface plasmon resonance biosensor. AB - This study demonstrates the first use of surface plasmon resonance (SPR) technology for the rapid, sensitive and label-free detection of whole B. anthracis spores. The approach involves the use of an SPR biosensor (Biacore 3000), and a monoclonal antibody which was raised against the B. anthracis spore (mAb 8G3). By means of subtractive inhibition assays, whole B. anthracis spores with concentrations as low as 10(4) colony-forming units (CFU) ml(-1) can be detected within 40 min, and other related Bacillus spores, even in high concentrations, can be differentiated from B. anthracis spores. PMID- 19305925 TI - Bio-electrospraying living Xenopus tropicalis embryos: investigating the structural, functional and biological integrity of a model organism. AB - Bio-electrosprays, a recently pioneered direct cell engineering approach, have been demonstrated to handle living cells including stem cells for the development of active specialized and unspecialized microenvironments. This electric field driven technique is currently undergoing vigorous development where the technique is racing towards possible clinical utility. Although this direct cell engineering approach has been elucidated to have no significant effects on the processed cells from a molecular level upwards, the technique needs to demonstrate its potential for use with whole organisms (multi-cellular systems). We believe this is mandatory for whole organisms such as model embryos; developing multi-cellular biological structures are sensitive systems and could possibly be prone to a wide range of embryological disruptions during their dynamic development, post-treatment. Therefore our studies presented herein have investigated the effects on embryos in terms of their structure, function and biological integrity post-bio-electrospraying in comparison to several controls. Our investigations demonstrate the absence of any detectable gross effects on the embryos from a genetic level upwards on post-treated embryos. In fact, these studies clearly elucidate no significant disruptions on the dynamic development of these treated embryos in comparison to those respective controls, thus validating the utility of bio-electrosprays for the careful handling of dynamically developing multi-cellular organisms. PMID- 19305926 TI - Use of simultaneous dual-probe microdialysis for the determination of pesticide residues in a jade plant (Crassula ovata). AB - A microdialysis application for the sampling of plant pesticide residues has been developed utilizing two probes with differing perfusate rates simultaneously inserted into symmetric components of a sampling system. A simple empirical equation is proposed to calculate analyte concentrations in a semi-solid matrix using two different dialysate concentrations. In vitro application yielded excellent correlations between the calculated and spiked concentrations. Subsequent in vivo application of this technique for the determination of pesticide distributions in leaves of a jade plant (Crassula ovata) indicated the utility of this approach in complex living matrices. PMID- 19305927 TI - Optimizing the detergent concentration conditions for immunoprecipitation (IP) coupled with LC-MS/MS identification of interacting proteins. AB - Immunoprecipitation (IP) coupled with LC-MS/MS is a widely used method in proteomics research to identify proteins and to study protein-protein interactions. IP techniques allow purification of proteins of interest and reduce sample complexity before introduction to the mass spectrometer. The effectiveness of IP experiments is an important factor for identification of proteins and protein-protein interactions. In this paper, a variety of IP conditions were studied systematically to improve IP-based protein interaction identification capabilities. Low concentration detergent (around 0.05% NP40/PBS) was found to improve IP effectiveness by decreasing non-specific binding. However, higher concentration detergent (e.g. 1% NP40/PBS) was detrimental. Furthermore, with lower protein concentrations, the IP system showed lower tolerance to detergent. For example, with a detergent concentration higher than 0.05% NP40/PBS, IP experiments were unsuccessful with low protein concentration (e.g. 0.28 microM ADH). In some cases the observed results were even worse than the results obtained without detergent. However, when the protein concentration was high (e.g. 1.12 microM ADH), this effect was not obvious and the high detergent (higher than 0.1%) experimental results were similar to those from low detergent concentration experiments (around 0.05%). Another application of this strategy to a more general system based on FLAG-Bacterial Alkaline Phosphatase (BAP) and anti FLAG antibody was also performed. These results also suggested that low detergent concentration (0.05% NP40) is helpful for IP experiments, especially for the experiments with low protein concentrations. Considering that in most real applications, the proteins of interest are usually present in low abundance, a low amount of detergent is recommended to be used. The optimized detergent concentration was determined to be 0.05% in these studies. However, the key result presented here illustrates that both detergent and protein concentrations should be carefully considered when one is trying to optimize IP prior to mass spectrometry experiments. PMID- 19305928 TI - Fourier transform infrared spectroscopic studies of T-cell lymphoma, B-cell lymphoid and myeloid leukaemia cell lines. AB - This paper presents Fourier transform infrared (FT-IR) spectroscopy to characterise spectral differences that distinguish cells derived from human T cell lymphoma, B-cell lymphoid, and myeloid leukaemia cell lines. This methodology is based on spectral measurements of major cellular biochemical constituents and multivariate spectral processing. Major spectral differences were observed in the 1800-900 cm(-1) 'fingerprint' spectral region. Bands in the averaged spectra for each cell line were assigned to major biochemical constituents including: proteins, lipids, carbohydrates and nucleic acids. Multivariate statistical analysis of the spectra was carried out to develop a classification model to discriminate the five cell types. The results show that FT-IR spectroscopy displays high sensitivity and specificity when discriminating between T-cell lymphoma, B-cell lymphoid, and myeloid leukaemia cells based on intrinsic biomolecular signatures. FT-IR spectroscopy in combination with multivariate statistical analysis provides an important insight into T-cell lymphoma, B-cell lymphoid, and myeloid leukaemia cell line identification. In conclusion, this paper demonstrates a potential for this technique to be used in developing a clinical tool for the detection and identification of haematological malignancies. PMID- 19305929 TI - Latent fingerprints analysis using tape-lift, Raman microscopy, and multivariate data analysis methods. AB - This paper describes the use of combined techniques, i.e. Raman spectral mapping, tape-lift, and multivariate data analysis, to extract chemical information of latent fingerprint and/or trace amounts of materials deposited in fingerprints. The tape-lift method was employed to lift trace particles, extrinsic materials, or sebum deposited on the finger of an individual after recent handling of such materials. The analysis of the tape-lifted materials was performed by Raman spectral mapping at a specific area. The collected mixture Raman spectra containing signals from lifting media and lifted materials was then deconvoluted using a powerful multivariate technique, namely band-target entropy minimization (BTEM). Three cases, i.e. a sebum-rich fingerprint after touching the forehead, a drug model comprising ibuprofen, L-arginine, and sodium bicarbonate, and an additive model comprising sucrose and aspartame were investigated. BTEM could recover all pure component spectra of both lifting media and tape-lifted materials. As such, all these test substances can be correctly identified using their unique pure Raman spectral signatures. In addition, the spatial distributions of all these identified components could also be determined. These combined three techniques hold promise as a new tool in forensic applications. PMID- 19305930 TI - Surface-adsorbed silver half-shells as a platform for surface-enhanced immunoassays; optimization through morphological control. AB - We have investigated the effectiveness of surface-adsorbed silver half-shells for inducing the surface-enhanced fluorescence phenomenon. Our simple structure consists of a dense monolayer of monodispersive latex sphere covered by thermally evaporated silver, some tens of nm thick. In order to increase its effectiveness as a platform for immunoassay, we explored three physical parameters; the diameter of the latex sphere, the deposition thickness and the adsorption density of the latex sphere. The maximum enhancement of 25.5 was achieved using the sphere with 56 nm diameter with 10 nm silver deposited. As for the adsorption density, the maximum fluorescence signal was achieved with a relatively sparse adsorption density, 12.5 per microm(2) vs. 105.9 per microm(2), corresponding to the full coverage. With the adsorption density also optimized, the enhancement was further increased by a factor of three. PMID- 19305931 TI - Comparative study of poly(styrene-sulfonate)/poly(L-lysine) and fibronectin as biofouling-preventing layers in dissolved oxygen electrochemical measurements. AB - Oxygen is a major actor of many physiological, biological and industrial processes; as such, its monitoring is of paramount importance. The effects of protein biofouling on dissolved oxygen measurements are described. The consequences of protein adsorption on electrode kinetics and mass transport were quantified using cyclic voltammetry, AC impedance and rotating disc amperometry, firstly in a solution of albumin, secondly in a complex protein suspension of liver tissue. The effect of membrane coatings poly(styrene-sulfonate)/poly(L lysine) and fibronectin were investigated. The relative significance on mass transport, surface diffusion and electrocatalysis were examined by comparing the experimental data for an outer-sphere redox couple, ruthenium hexaammine (III)/(II), with the physiologically significant electrocatalytic O(2) reduction reaction. PMID- 19305932 TI - Non-enzymatic glucose detection using as-prepared boron-doped diamond thin-film electrodes. AB - Electrochemical oxidation of glucose in alkaline solution at as-prepared boron doped microcrystalline diamond (BDMD) and nanocrystalline diamond (BDND) thin film electrodes is investigated by cyclic voltammetry. The results demonstrate that glucose can be directly oxidized at as-prepared boron-doped diamond (BDD) thin-film electrodes and the curve of the negative scan traces onto the positive scan. The effect of sodium hydroxide concentration on the response of glucose is also studied in the range of 0.02-0.6 M and the optimum concentration of sodium hydroxide is found to be 0.1 M. The voltammetric signal of glucose and the mixture of ascorbic acid (AA) and uric acid (UA) can be observed well-separated at as-prepared BDD thin-film electrodes in 0.1 M sodium hydroxide solution. The peak current is proportional to the glucose concentration in the range 0.25-10 mM with a correlation coefficient of 0.9993 in the presence of AA and UA. Furthermore, the experiment results also show that the non-enzymatic glucose sensor based on as-prepared BDD thin-film electrodes has high sensitivity, good reproducibility and stability. PMID- 19305933 TI - Magnetic bead-based DNA hybridization assay with chemiluminescence and chemiluminescent imaging detection. AB - Simple and sensitive chemiluminescence (CL) and CL imaging methods have been developed for the magnetic bead-based DNA hybridization assay. The assay relies on the high sensitivity and long stable light signal of the CL system in which horseradish peroxidase (HRP) catalyzes the luminol-H(2)O(2) reaction with para iodophenol (PIP) as the enhancer. In this protocol, a sandwich DNA hybridization is performed by mixing the target DNA with the magnetic bead-captured DNA and the biotinylated reporter DNA, followed through the biotin-streptavidin reaction with conjugated HRP, and then the conjugated HRP is determined by the CL system. The proposed CL protocol is suitable for for the detection of sequence-specific DNA related to the avian influenza A H1N1 virus at levels as low as 10 amol, and the CL imaging detection has a similar sensitivity. The sensitivities of the proposed methods with the HRP label are better than most of the metal nanoparticle-based methods, and are comparable with that of utilizing amplified techniques for DNA hybridization detection. In addition, the perfectly complementary DNA sequences and the single-base mismatched DNA sequences can be better distinguished by a thermally-stringent hybridization and washing steps. So, the proposed CL method can offer great promise for single-nucleotide polymorphism (SNP) analysis. Moreover, the proposed method may have significant potential for the simultaneous detection of various DNA sequences when different capture DNA sequences and reporter DNA sequences are used in a microarray. PMID- 19305934 TI - Should studies of risk factors for musculoskeletal disorders be stratified by gender? Lessons from the 1998 Quebec Health and Social Survey. AB - OBJECTIVES: Several studies have reported male-female differences in the prevalence of symptoms of work-related musculoskeletal disorders (MSD), some arising from workplace exposure differences. The objective of this paper was to compare two strategies analyzing a single dataset for the relationships between risk factors and MSD in a population-based sample with a wide range of exposures. METHODS: The 1998 Quebec Health and Social Survey surveyed 11 735 respondents in paid work and reported "significant" musculoskeletal pain in 11 body regions during the previous 12 months and a range of personal, physical, and psychosocial risk factors. Five studies concerning risk factors for four musculoskeletal outcomes were carried out on these data. Each included analyses with multiple logistic regression (MLR) performed separately for women, men, and the total study population. The results from these gender-stratified and unstratified analyses were compared. RESULTS: In the unstratified MLR models, gender was significantly associated with musculoskeletal pain in the neck and lower extremities, but not with low-back pain. The gender-stratified MLR models identified significant associations between each specific musculoskeletal outcome and a variety of personal characteristics and physical and psychosocial workplace exposures for each gender. Most of the associations, if present for one gender, were also found in the total population. But several risk factors present for only one gender could be detected only in a stratified analysis, whereas the unstratified analysis added little information. CONCLUSIONS: Stratifying analyses by gender is necessary if a full range of associations between exposures and MSD is to be detected and understood. PMID- 19305935 TI - Adverse life events among community-dwelling persons aged 65-70 years: gender differences in occurrence and perceived psychological consequences. AB - OBJECTIVES: To describe the occurrence of selected adverse life events in young old men and women, as well as their perceived psychological consequences. METHODS: In 2005, 1,422 participants in the Lausanne Cohort 65+ study, born in 1934-1938, self-reported whether they experienced any of 26 life events during the preceding year. Most participants (N = 1,309, 92%) completed the geriatric adverse life events scale during a face-to-face interview, by rating the level of stress associated with each event, as well as its impact on their psychological well-being. RESULTS: Overall, 72% of the participants experienced at least one of the 26 events in the preceding year (range 1-9). Disease affecting the respondent (N = 525) or a close relative (N = 276) was most frequent, as well as the death of a friend or non-close relative (N = 274). Women indicated a higher frequency of events (mean 2.1 vs. 1.7 events, P < 0.001), as well as a higher level of stress and a stronger negative impact on well-being than men. In multivariate analyses adjusting for self-rated health, depressive symptoms and comorbidity, female gender remained significantly associated with the level of stress and negative impact on psychological well-being. CONCLUSION: This exploratory study shows that several types of adverse life events frequently occur at age 65-70, with gender differences both in the frequency of reporting and consequences of these events. However, information on this topic is limited and studies based on different populations and designs are needed to better understand the impact of such events. PMID- 19305936 TI - Evidence of the effectiveness of a specialist vocational intervention following first episode psychosis: a naturalistic prospective cohort study. AB - BACKGROUND: Employment rates among people with severe mental illness are low and work has beneficial effects on mental health. There is now good evidence of the effectiveness of a specialist vocational intervention (supported employment) in people with schizophrenia. However, the potential benefits of modifying this model for use in first episode psychosis cohorts remain relatively untested. METHODS: The aim of our study was to evaluate the effectiveness of a specialist vocational intervention in aiding vocational recovery following the onset of first episode psychosis. In a naturalistic prospective cohort study, 114 first episode psychosis service users were followed up during 12 months of engagement with an early intervention service; 44 resident in an area where a vocational intervention was available and 70 in an area where it was not. RESULTS: The main finding in our study was that having access to the specialist vocational intervention was a statistically significant independent predictor of vocational recovery during 12 months of follow-up (after adjusting for confounders). Service users who had access to the intervention had odds of achieving vocational recovery 3.53 times greater than those who did not (OR = 3.53, 95% CI = 1.25 10.00). CONCLUSION: This study provides further preliminary evidence of the effectiveness of a specialist vocational intervention following first episode psychosis. This is an important outcome from the perspective of service users and clinicians alike (as well as having wider societal value). Other important predictors of vocational recovery cannot be modified by the time a first episode psychosis emerges. PMID- 19305937 TI - Attitudes to people with mental disorders: a mental health literacy survey in a rural area of Maharashtra, India. AB - INTRODUCTION: People with mental disorders experience discrimination as a consequence of stigmatising attitudes that are largely socio-culturally constructed. Thus, there is a need to understand local contexts in order to develop effective programs to change such attitudes. We undertook a mental health literacy survey in rural Maharashtra, India, prior to developing a mental health training program for village health workers (VHWs) in a primary health care setting. METHODS: A cross-sectional mental health literacy survey was undertaken in late 2007, which involved interviewer-administration of a questionnaire to 240 systematically sampled community members, and 60 purposively sampled VHWs. Participants were presented with two vignettes describing people experiencing symptoms of mental disorders (depression, psychosis), and were asked about attitudes towards, and desired social distance from, the people in the vignettes (the latter being a proxy measure for stigma). Linear regression modelling was undertaken to identify predictors of social distance. RESULTS: Although the community was relatively accepting of people with mental disorders, false beliefs and negative attitudes were still evident. Desired social distance was consistently greater for the person depicted in the psychosis vignette compared to the depression vignette. For both vignettes, the main predictor of greater social distance was perceiving the person as dangerous, and the predictors of reduced social distance were being a VHW, and seeing the problem as a sign of personal weakness. For depression, believing the cause to be family tensions also reduced social distance. For psychosis, labelling the disorder as a mind/brain problem, and believing the cause to be lack of control over life or genetic factors increased social distance. The vast majority did not agree that the problems experienced in the vignettes were 'a real medical illness'. CONCLUSION: Promoting bio-medical explanations for mental disorders in this setting may exacerbate discriminatory attitudes. Provision of contextually relevant mental health training for the VHWs so that they are able to communicate, model and shape more positive attitudes is the next step. PMID- 19305938 TI - Dimensional, non-taxonic latent structure of psychotic symptoms in a student sample. AB - INTRODUCTION: We examined the construct of psychosis using the self-report instrument CAPE (Community Assessment of Psychic Experiences) in a sample of 1,323 students. MATERIALS AND METHODS: Taxometric analysis was carried out using Taxometric Programs for the R Computing Environment. The MAXCOV CCFI was 0.34, indicating a dimensional latent structure. All other taxometric analysis yielded very similar results indicative of dimensional structure. CONCLUSION: In this study, using powerful analytic techniques designed expressly for the purpose, i.e. taxometric analysis, the latent construct of psychosis in a sample of young students appeared to be consistent with a dimensional, non-taxonic latent structure. PMID- 19305939 TI - Effect of fire on pools of mercury in forest soil, Central Europe. AB - In year 2006, 17.9 ha of forest was burned during a forest fire at the Bohemian Switzerland National Park found in northern part of Czech Republic (CR), central Europe. Complete combustion of organic soil (4,039 t) on the burned area caused volatilization of 1.34 +/- 0.07 kg of Hg. Thus Hg emissions due to fire amounted to 75.1 g ha(-1). The average burned forested areas in CR for the period 2000 2006 were reported at 356 ha with estimated Hg emissions at 26.7 kg year(-1), while the average anthropogenic emissions in the same period amounted to 3 t year(-1). Thus estimated mean emissions of Hg from burned forest soil in the period 2000-2006 reached 1% of the annual anthropogenic Hg emissions. PMID- 19305940 TI - Pre- and post-operative work-up in patients affected by early cervical cancer and eligible for fertility-sparing treatment: role of MRI with saline hydrocolpos. AB - Nowadays young women affected by early invasive uterine cervical cancer (stage IA2-IB1) may be offered a fertility-sparing treatment: the radical trachelectomy with pelvic lymph node dissection. This procedure consists in surgical removal of cervix uteri, proximal parametrial tissue, and vaginal cuff. The morphology and the functions of corpus uteri are preserved. Women candidates for trachelectomy must be closely selected. Gynecologist oncologist needs an imaging modality that can accurately value the tumoral diameter and which can demonstrate proximal extension of tumor to ensure surgical clear resection margins (especially the cranial one). Magnetic resonance imaging (MRI) is a very useful examination in pre-operative study of women affected by early cervical cancer. The aim of this study is to evaluate the role of MRI with hydrocolpos about pre- and post operative work-up in women eligible for fertility-sparing treatment. PMID- 19305941 TI - Detection of recurrent hernia and intraabdominal adhesions following incisional hernia repair: a functional cine MRI-study. AB - BACKGROUND: To non-invasively identify incisional hernia repair implanted synthetic meshes with MRI, and also focusing on the evaluation of postsurgical complications such as adhesions. METHODS: A total of 43 patients underwent either laparoscopic intraperitoneal onlay-mesh or open abdominal wall repair using preperitoneal layers. The patients were examined using a true-fast-imaging-with steady-state-precession (trueFISP)-sequence in transverse/sagittal orientation with a section-by-section dynamic depiction of induced visceral slide. A 9 segment-abdominal-map was used to document the adhesion location/type. The MR images were analysed regarding hernia relapse, layer-morphology, rectus-abdominis muscle-condition, and abdominal wall mobility. In 12 patients pre- and postsurgery-MRI was performed. RESULTS: Time range between surgery and examination was 6-36 months. In all laparoscopy-patients the meshes were identified. For open surgery the mesh was not visualized in 20, but was seen in 6 cases. A total of 11 cases showed a recurrent hernia. Seventy intraabdominal adhesions were detected. Fifteen patients had restricted mobility. 20 patients showed an rectus-abdominis-muscle-asymmetry. Comparing pre- and post-op-MRI, 6 out of 8 patients with open repair showed thick scar-plaques. Three patients with open repair had new adhesion-formations postoperatively. CONCLUSION: Functional cine MRI is suitable for follow-up studies in patients after hernia repair to detect and evaluate the implanted meshes. Typical complications like intestinal adhesions and abdominal wall dysmotility can be assessed as well. PMID- 19305942 TI - In vitro assembly of nucleocapsid-like particles from purified recombinant capsid protein of dengue-2 virus. AB - The capsid protein is one of the three structural proteins of flaviviruses and is the building block of the nucleocapsid. It has also a predominant role in the replication of dengue virus. To obtain nucleocapsid-like particles from recombinant dengue-2 capsid protein produced in E. coli, a purification process using cation exchange chromatography was established. The purified protein exhibited a molecular mass corresponding to a dimer; therefore, similar to that reported for alphaviruses, an in vitro assembly reaction using single-stranded DNA was performed. In all cases, particles were obtained independently of the specificity and the length of the oligonucleotides used. The present work is the first report of in vitro assembly of the recombinant dengue capsid protein, which could constitute a powerful tool in the development of vaccine candidates. PMID- 19305943 TI - Biogenic methane production in formation waters from a large gas field in the North Sea. AB - Methanogenesis was investigated in formation waters from a North Sea oil rimmed gas accumulation containing biodegraded oil, which has not been subject to seawater injection. Activity and growth of hydrogenotrophic methanogens was measured but acetoclastic methanogenesis was not detected. Hydrogenotrophic methanogens showed activity between 40 and 80 degrees C with a temperature optimum (ca. 70 degrees C) consistent with in situ reservoir temperatures. They were also active over a broad salinity range, up to and consistent with the high salinity of the waters (90 g l(-1)). These findings suggest the methanogens are indigenous to the reservoir. The conversion of H(2) and CO(2) to CH(4) in methanogenic enrichments was enhanced by the addition of inorganic nutrients and was correlated with cell growth. Addition of yeast extract also stimulated methanogenesis. Archaeal 16S rRNA gene sequences recovered from enrichment cultures were closely related to Methanothermobacter spp. which have been identified in other high-temperature petroleum reservoirs. It has recently been suggested that methanogenic oil degradation may be a major factor in the development of the world's heavy oils and represent a significant and ongoing process in conventional deposits. Although an oil-degrading methanogenic consortium was not enriched from these samples the presence and activity of communities of fermentative bacteria and methanogenic archaea was demonstrated. Stimulation of methanogenesis by addition of nutrients suggests that in situ methanogenic biodegradation of oil could be harnessed to enhance recovery of stranded energy assets from such petroleum systems. PMID- 19305944 TI - Thermovorax subterraneus, gen. nov., sp. nov., a thermophilic hydrogen-producing bacterium isolated from geothermally active underground mine. AB - A thermophilic, rod-shaped, motile, Gram-positive, spore-forming bacterium strain 70B(T) was isolated from a geothermally active underground mine in Japan. The temperature and pH range for growth was 50-81 degrees C (optimum 71 degrees C) and 6.2-9.8 (optimum pH 7-7.5), respectively. Growth occurred in the presence 0 2% NaCl (optimum 1% NaCl). Strain 70B(T) could utilize glucose, fructose, mannose, mannitol, pyruvate, cellobiose and tryptone as substrates. Thiosulfate was used as electron acceptor. Major whole-cell fatty acids were iso-C(15:0), C(16:0) DMA (dimethyl acetal), C(16:0) and anteiso-C(15:0). The G+C mol% of the DNA was 44.2%. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the closest relatives of strain 70B(T) were Thermosediminibacter oceani DSM 16646(T) (94% similarity) and Thermosediminibacter litoriperuensis DSM 16647 (93% similarity). The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain 70B(T) represents a novel species in a new genus, for which the name Thermovorax subterraneus gen. nov., sp. nov. is proposed. The type strain of Thermovorax subterraneus is 70B(T) (=DSM 21563 = JCM 15541). PMID- 19305945 TI - Systemic mastocytosis presenting as osteoporosis--a case report. AB - Mastocytosis is a clonal disorder of the mast cell and its precursor cells, and is characterised by proliferation and accumulation of mast cells within various organs, most commonly the skin. Systemic mastocytosis is a rare but well recognised cause of secondary osteoporosis, accounting for about 1.25% of cases. The pathophysiological mechanism is probably multifactorial, including increased osteoclastic activity, and a direct effect of mast cell mediators like histamine, heparin, tryptase and cytokines. Here, we report the case of a middle-aged male patient with osteoporotic vertebral fractures as a rare presenting manifestation of systemic mastocytosis. In summary, systemic mastocytosis, although rare, should be considered as a cause in patients with idiopathic osteoporosis. PMID- 19305946 TI - Preconditioning and its clinical potential. AB - Preconditioning is emerging as a simple, safe and highly effective means of attenuating local and systemic effects of medical and surgical insult. Its enormous potential has not yet been harnessed and ongoing work will continue to bring it to the fore. This article covers the history, development and future clinical potential of preconditioning with particular regard to surgical insult. PMID- 19305947 TI - Two cases of hemichorea-hemiballism with nonketotic hyperglycemia: a new point of view. AB - Hemichorea-hemiballism (HCHB) is an usually continuous, nonpatterned, involuntary movement disorder caused by basal ganglia dysfunction, commonly due to a vascular lesion, described in nonketotic hyperglycemic patients. Particular computed tomography and magnetic resonance imaging findings have been described. The pathogenic mechanism of chorea arising during hyperglycemia and the nature of neuroimaging findings are unclear. In this paper we describe two elderly women with onset of HCHB during a hyperglycemic episode. The symptoms persisted in one of them after recovery of normal glycemia. The pathophysiological mechanism of the disease is discussed in the light of clinical and neuroradiological follow up. PMID- 19305948 TI - School opportunities and physical activity frequency in nine year old children. AB - OBJECTIVE: To examine the association between physical activity (PA) opportunities at school and participation in PA outside of school physical education (PE) classes among 9 year old children. METHODS: Data were obtained in a representative sample of 1 267 students nested within 69 schools who completed questionnaires; principals provided data on schools. Students' PA frequency was defined as all reported episodes of past week PA, excluding PE class. Schools were categorized as "high-opportunity" if they provided > or = 4 of the following opportunities: PE class > or = 2 times/week, extracurricular sports animator, gymnasium, swimming pool, schoolyard equipped for games, and > or = 1 outdoor sports playing fields. Multilevel modelling techniques were used to investigate the association between school-level PA opportunity and student-level PA frequency. RESULTS: No main effect of school-level PA opportunities on PA frequency was observed. However, overweight boys attending high-opportunity schools were significantly more active than those attending low-opportunity schools. CONCLUSIONS: The association between PA frequency and school-level PA opportunity differs by sex and weight status. Overweight boys in particular may benefit from health promotion strategies providing greater opportunities for school PA. PMID- 19305950 TI - Introduction of tau mutation into cultured Rat1-R12 cells by gene targeting, using recombinant adeno-associated virus vector. AB - We aim to develop a cultured cell model, to serve as a system with which the altered circadian phenotypes produced by the clock gene variations could be studied in vitro. Tau mutation, which shortens the circadian period of hamsters and mice, was introduced into the CK1epsilon locus of cultured Rat1-R12 cells by gene targeting mediated by a recombinant adeno-associated virus (rAAV) vector. After transduction of Rat1-R12 cells with rAAV, about 0.14% of the drug-resistant cells underwent gene targeting at CK1epsilon locus. Of the three clones isolated, only one carried the targeted allele of tau mutation and two carried the targeted wild-type allele. The clone with the targeted tau mutant allele exhibited a significantly shorter circadian period compared to the clone with targeted wild type allele. rAAV-mediated gene targeting in cultured somatic cells is a convenient and powerful tool for analyzing the phenotypic outcome of clock gene variations, and for elucidating the pathogenesis of the disorders associated with abnormal circadian rhythmicity. PMID- 19305951 TI - Immunohistochemical study of postnatal neurogenesis after whole-body exposure to electromagnetic fields: evaluation of age- and dose-related changes in rats. AB - It is well established that strong electromagnetic fields (EMFs) can give rise to acute health effects, such as burns, which can be effectively prevented by respecting exposure guidelines and regulations. Current concerns are instead directed toward the possibility that long-term exposure to weak EMF might have detrimental health effects due to some biological mechanism, to date unknown. (1) The possible risk due to pulsed EMF at frequency 2.45 GHz and mean power density 2.8 mW/cm(2) on rat postnatal neurogenesis was studied in relation to the animal's age, duration of the exposure dose, and post-irradiation survival. (2) Proliferating cells marker, BrdU, was used to map age- and dose-related immunohistochemical changes within the rostral migratory stream (RMS) after whole body exposure of newborn (P7) and senescent (24 months) rats. (3) Two dose related exposure patterns were performed to clarify the cumulative effect of EMF: short-term exposure dose, 2 days irradiation (4 h/day), versus long-term exposure dose, 3 days irradiation (8 h/day), both followed by acute (24 h) and chronic (1 4 weeks) post-irradiation survival. (4) We found that the EMF induces significant age- and dose-dependent changes in proliferating cell numbers within the RMS. Our results indicate that the concerns about the possible risk of EMF generated in connection with production, transmission, distribution, and the use of electrical equipment and communication sets are justified at least with regard to early postnatal neurogenesis. PMID- 19305952 TI - Comparison of human post-embryonic, multipotent stem cells derived from various tissues. AB - Multipotent stem cells were isolated from human fetal heart, liver, muscle, lung, derma, kidney, and adipose tissue, and then analyzed for their characteristics and function. Cells with characteristics similar to bone marrow-derived post embryonic multipotent stem cells can be selected and cultured from tissues other than bone marrow. This may then help explain the "stem cell plasticity" found in multiple human tissues. PMID- 19305953 TI - Construction, expression and characterization of fusion enzyme from Arthrobacter oxydans dextranase and Klebsiella pneumoniae amylase. AB - An artificial fusion protein of Arthrobacter oxydans dextranase and Klebsiella pneumoniae alpha-amylase was constructed and expressed in Escherichia coli. Most of the expressed protein existed as an insoluble fraction, which was solubilized with urea. The purified fusion enzyme electrophoretically migrated as a single protein band; M = 137 kDa, and exhibited activities of both dextranase (10.8 U mg(-1)) and amylase (7.1 U mg(-1)), which were lower than that of reference dextranase (13.3 U mg(-1)) and alpha-amylase (103 U mg(-1)). The fusion enzyme displayed bifunctional enzyme activity at pH 5-7 at 37 degrees C. These attributes potentially make the fusion enzyme more convenient for use in sugar processing than a two-enzyme system. PMID- 19305954 TI - Interpretation of pulmonary function test: issues and controversies. AB - Pulmonary function testing (PFT) serves many purposes in clinical practice, and in contrast to other laboratory measures, PFT results are often provided with a clinical interpretation. PFT interpretation depends on the clinical context, and multiple challenges influence PFT interpretation. Overall, the goal of PFT interpretation is to distinguish normal from abnormal, and this is affected by the selection of reference standards, as well as the arbitrary albeit not necessarily irrational choice of cut-off values. Controversies regarding PFT analysis may lead to important differences in interpretation. In this article, issues associated with the selection of reference standards are discussed, followed by a review of the controversies related to PFT interpretation in the diagnosis of obstructive ventilatory defect, restrictive ventilatory defect, gas transfer defect, and flow-volume loop abnormalities. Given the challenges facing PFT interpretation, no single interpretative algorithm is sufficient; rather, PFT interpretation requires a comprehensive approach including consideration of the clinical context, laboratory methodology, and reference standards and an understanding of the consequences of a normal or abnormal designation. PMID- 19305956 TI - [Causes of leg ulcers. Analysis of the data from a dermatologic wound care center]. AB - The etiology of chronic leg ulcers is very heterogeneous. The primary aim of our investigation was to analyze the etiology of leg ulcers in patients treated in our wound care center over a period of 5 years. In this open retrospective study, data of 354 patients was analyzed. We found a venous leg ulcer in 203 (57%), and in 52 (15%) patients a combined arterio-venous induced leg ulcer. The third most common cause was leg ulcers due to different types of vasculitis in 47 (13%) patients. An exclusively arterial induced leg ulcer was diagnosed in 13 (3.7%) patients. Overall 16 different etiologies were identified. In contrast to the few comparable other investigations, we found a higher incidence of immunologic und infectious causes for leg ulcers in our patients. Even if our results do not allow a representative overview about the prevalence of leg ulcer in Germany, they demonstrate the wide spectrum of leg ulcer causes in a dermatologic wound care centre. PMID- 19305957 TI - [Therapy of bronchial asthma according to the guidelines]. PMID- 19305958 TI - [Drug therapy during pregnancy]. AB - Most acute and chronic diseases do not require termination of a pregnancy nor is a pregnancy a reason to withhold necessary drug therapy. As many pregnancies are unplanned, any drug treatment during reproductive age should consider the occurrence of a pregnancy. Wherever possible, only medicinal drugs with sufficient documentation in pregnant women not indicating developmental toxicity should be chosen. There are acceptable drugs for the majority of diseases, although many of them are labeled as contraindicated during pregnancy. This article provides an overview on prenatal risks of ACE-inhibitors, AT II-receptor antagonists, antiepileptics, SSRI, atypical neuroleptics, lithium, coumarin anticoagulants, retinoids, selected antibiotics, immunomodulatory drugs and provides treatment recommendations for the most common diseases. PMID- 19305959 TI - [Differential hypertensive therapy according to the guidelines]. AB - The goal of antihypertensive therapy is to lower blood pressure and, by doing so, to decrease cardiovascular risk. Life style changes and drugs are available for the treatment of hypertensive patients. In order to reach the target blood pressure, most patients with hypertension need drug treatment in addition to life style changes. In all hypertensive patients, the target blood pressure is <140/90 mmHg. In patients with diabetes mellitus, with chronic renal failure as well as in patients with complications of hypertension and, thereby a very high cardiovascular risk, the target blood pressure is <130/80 mmHg. Diuretics, beta blockers, calcium antagonists, ACE-inhibitors and angiotensin receptor blockers are the drugs of first choice in the treatment of hypertension. The selection among these drug classes has to consider probable side effects as well as accompanying diseases and complications of hypertension. One should also take into account that most of the beneficial effects of antihypertensive therapy is due to the decrease of blood pressure per se and that the majority of hypertensive patients require the combination of two or more antihypertensive drugs in order to reach the target blood pressure. PMID- 19305960 TI - Femoral component positioning in hip resurfacing with and without navigation. PMID- 19305961 TI - Gait and stair function in total and resurfacing hip arthroplasty: a pilot study. AB - Standard total hip arthroplasty (THA) is the established surgical treatment for patients older than 65 years with progressive osteoarthritis but survivorship curves wane in patients younger than 50. Resurfacing hip arthroplasty (RHA) is an alternative for younger, active patients reportedly providing superior range of motion. Quantitative investigation of functional recovery following arthroplasty may elucidate limitations that aid in device selection. Although limited long term kinematic data are available, the early rate of recovery and gait compensations are not well described. This information may aid in refining rehabilitation protocols based on limitations specific to the implant. We presumed hip motion and forces for subjects receiving RHA are more similar to age matched controls during physically demanding tasks, such as stair negotiation, at early time points than those for THA. In a pilot study, we quantified walking and stair negotiation preoperatively and 3 months postoperatively for seven patients with RHA (mean age, 49 years), seven patients with standard THA (mean age, 52 years), and seven age-matched control subjects (mean age, 56 years). Although both treatment groups demonstrated trends toward functional recovery, the RHA group had greater improvements in hip extension and abduction moment indicating typical loading of the hip. Further investigation is needed to determine if differences persist long term or are clinically meaningful. PMID- 19305962 TI - [Disorders of the patellofemoral joint induced by weight training]. AB - The patellofemoral joint is stressed strongly during weight training. There are different individual preconditions that influence the stress distribution. Forces on tendons and cartilage are a function of angle and exercise. The most common disorders are tendinopathies and cartilage damage. Patellofemoral malalignment, high loads and overuse, uncontrolled exercises and steroids represent risk factors for injuries. Individual training concepts with controlled exercises that reduce peak loads are desirable. There is a wide scope of therapeutic options ranging from antiphlogistic therapy to the reconstruction of tendons and cartilage. PMID- 19305963 TI - [Minimally invasive fixation of a sacral bilateral fracture with lumbopelvic dissociation]. AB - This case report describes our technique of percutaneous sacroiliac screw fixation of a bilateral sacral fracture with spinopelvic dissociation. It is based on the description of an iliosacroiliac bony corridor delimited by the following landmarks: the sacral ala in a superior-anterior direction, the first sacral foramen on both sides in an inferior-posterior direction, and the sacral channel posteriorly. The described operating method, which uses a 6.5-mm parallel drill sleeve, allows the safe and strictly transversal positioning of a 7.3-mm screw on each side with the threads interlocking. It provides a separate and fracture-adapted compression of the screws. The interlocking SI screw threads increase the pull-out strength. Operating time and radiation dose can be reduced significantly by this method. PMID- 19305964 TI - What's the buzz? Ultrasonic and sonic warning signals in caterpillars of the great peacock moth (Saturnia pyri). AB - Caterpillars have many natural enemies and, therefore, have evolved a diversity of antipredator strategies. Most research focuses on those strategies (crypsis, countershading, and warning coloration) targeting visually guided predators. In contrast, defensive sounds, although documented for more than a century, have been poorly studied. We report on a novel form of sound production--chirping--in caterpillars of the common European Great Peacock moth (Saturnia pyri). Chirps are broadband, with dominant peaks ranging between the sonic (3.7 kHz) and ultrasonic (55.1 kHz) and are generated by a rapid succession of mandibular "tooth strikes." Chirp trains are induced by simulated predator attacks and precede or accompany the secretion of a defensive chemical from integumental bristles, supporting our hypothesis that these sounds function in acoustic aposematism. We propose that these caterpillars generate multimodal warning signals (visual, chemical, and acoustic) to target the dominant sensory modalities of different predators, including birds, bats, and invertebrates. PMID- 19305965 TI - Ambulatory blood pressure measurements are related to albumin excretion and are predictive for risk of microalbuminuria in young people with type 1 diabetes. AB - AIMS/HYPOTHESIS: The relationship between BP and microalbuminuria in young people with type 1 diabetes is not completely clear. As microalbuminuria is preceded by a gradual rise in albumin excretion within the normal range, we hypothesised that ambulatory BP (ABP) may be closely related to albumin excretion and progression to microalbuminuria. METHODS: ABP monitoring (ABPM) was performed in 509 young people with type 1 diabetes (age median [range]: 15.7 [10.7-22.6] years) followed with annual assessments of three early morning urinary albumin:creatinine ratios (ACRs) and HbA(1c). Systolic BP (SBP) and diastolic BP (DBP) and the nocturnal fall in BP were analysed in relation to ACR. RESULTS: All ABPM variables were significantly related to baseline log(10) ACR (p < 0.001). After the ABPM evaluation, 287 patients were followed for a median of 2.2 (1.0-5.5) years. ABP at baseline was independently related to mean ACR during follow-up. Nineteen initially normoalbuminuric patients developed microalbuminuria after 2.0 (0.2 4.0) years and their baseline daytime DBP was higher than in normoalbuminuric patients (p < 0.001). After adjusting for baseline ACR and HbA(1c), there was an 11% increased risk of microalbuminuria for each 1 mmHg increase in daytime DBP. Forty-eight per cent of patients were non-dippers for SBP and 60% for DBP; however, ACR was not different between dippers and non-dippers and there were no differences in the nocturnal fall in BP between normoalbuminuric and future microalbuminuric patients. CONCLUSIONS/INTERPRETATION: In this cohort of young people with type 1 diabetes, ABP was significantly related to ACR, and daytime DBP was independently associated with progression to microalbuminuria. Increasing albumin excretion, even in the normal range, may be associated with parallel rises in BP. PMID- 19305966 TI - [Current diagnosis and therapy of osteoporosis on the basis of "European guidance 2008"]. AB - Osteoporosis is one of the 10 most important and widespread global diseases. In Germany alone the cost of osteoporosis runs into many billions of Euros. However, it should be noted that in the past 15 years great progress has been made both in diagnosis and in the development of new medications, and this has changed the general perception of and attitude to osteoporosis. It is now taken very seriously and recognised as a national and global disorder which is both preventable and treatable. In spite of this progress, in Europe and especially in Germany, osteoporosis remains an underdiagnosed and undertreated disease. In Germany, only about 10%-15% of patients with manifest osteoporosis are properly treated. However, in addition to national guidelines, there is now a new edition of the "European Position Paper for the Diagnosis and Management of Osteoporosis". This provides physicians treating osteoporosis patients with additional information and therefore more confidence. PMID- 19305967 TI - [Science and practice--a good mixture]. PMID- 19305968 TI - [Pediatric osteomyelitis]. AB - Bacterial osteomyelitis in children must be diagnosed quickly and requires immediate and adequate antibiotic treatment. Surgical interventions may be necessary. Infants as well as immunodeficient patients suffer more often from hematogenic bone infections than immunocompetent patients. According to recent findings, autoinflammatory nonbacterial osteitis is more probable in immunocompetent patients in good general condition and should always be considered as a differential diagnosis. Diagnostic and therapeutic approaches are presented when childhood osteomyelitis is suspected. PMID- 19305969 TI - Arthroscopic view of transient synovitis of the hip joint: a case report. AB - We present a case of a 13-year-old male patient with hip pain and limp. Inconclusive investigations and persistent symptoms resulted in the patient undergoing hip arthroscopy. At arthroscopy, a large focal cartilaginous defect of the femoral head with synovitis was seen and synovial biopsy confirmed the diagnosis. Arthroscopic debridement of the lesion, synovectomy and non-weight bearing for 6 weeks resulted in relief of symptoms. Transient synovitis is a common diagnosis in young children, but the extent of damage to the femoral head has not been previously described. Transient synovitis may not always respond to symptomatic treatment and hip arthroscopy may be useful in detecting the actual lesion and plan the treatment. PMID- 19305970 TI - The effect of intra-operative knee flexion angle on determination of graft location in the anatomic double-bundle anterior cruciate ligament reconstruction. AB - Graft tunnel placement is the factor with most influence on the outcome of double bundle anterior cruciate ligament (ACL) reconstruction. However the final decision for the graft location has to be decided subjectively under arthroscopy, and can be misplaced due to the effect of the knee flexion angle. The displacement of the estimated placement by surgeons from the ACL anatomical attachment is due to the knee's differing knee flexion angle. Eight cadaveric knees and an electromagnetic position recording system were employed. After digitizing the anatomical location of AM and PL bundle center, four experienced surgeons estimated the graft placement repeatedly at 70 degrees , 90 degrees and 110 degrees of knee flexion. The displacements between these two positions were calculated and analyzed separately in antero-posterior and disto-proximal directions. The displacements of the estimated AM bundle placements were 4.7 +/- 3.4 mm at 70 degrees , 4.3 +/- 2.2 mm at 90 degrees , and 6.0 +/- 2.6 mm at 110 degrees , while those of the PL bundle were 4.0 +/- 2.2 mm at 70 degrees , 3.4 +/ 1.9 mm at 90 degrees , and 4.2 +/- 2.5 mm at 110 degrees . The best results were obtained at 90 degrees of knee flexion. Additionally, the estimated placements for both AM and PL bundle were located more distally as the flexion angle increased. Our results imply that the knee should be set at 90 degrees when determining the graft placement in double-bundle reconstruction to prevent misplacement of the graft usually in a disto-proximal direction. PMID- 19305971 TI - Reliability testing of a new device to measure tibial rotation. AB - The purpose of this study was to evaluate the reliability of a new developed device to measure tibial rotation, the Rotameter. Thirty healthy subjects (15 males, 15 females, 24 years) were examined with the Rotameter measurement device. External and internal rotation was performed at an applied torque of 5, 10 and 15 Nm by two independent examiners in order to test the inter-observer reliability. The patients were measured again after a mean of 31 +/- 43 days by the same examiners to test the intra-observer reliability. Statistical analysis was performed using the intra-class correlation coefficient. The Pearson Correlation coefficient was used to compare the measurements of the left with the right side of the participants. In the measurements, a high inter- and intra-observer reliability was found at 5, 10 and 15 Nm of applied torque for the external rotation, internal rotation and the rotational range (internal + external rotation). Comparison of the left and the right knee of the same participant also revealed high correlations in the Pearson correlation coefficient at all applied torques. In conclusion, the Rotameter testing device for the measurement of tibial rotation showed a high inter-observer and intra-observer reliability. It is easy to perform and might be used in a wide field as a non-invasive instrument to objectively determine rotational stability and to investigate the restoration of the rotational stability after surgical procedures. PMID- 19305972 TI - Comparative evaluation of the tendon-bone interface contact pressure in different single- versus double-row suture anchor repair techniques. AB - The aim of the study was to evaluate the time zero contact pressure over a defined rotator cuff footprint using different repair and stitch techniques in an established sheep model. Forty fresh-frozen sheep shoulders were randomly assigned to five repair groups: single-row repair using simple stitches (SRA-s), single-row repair using horizontal mattress stitches (SRA-m), and single-row repair using arthroscopic Mason-Allen stitches (SRA-ama). Double-row repair was either performed with a combination of simple and horizontal mattress stitches (DRA-sm) or with arthroscopic Mason-Allen/horizontal mattress stitches (DRA amam). Investigations were performed using a pressure-sensitive film system. The average contact pressure and pressure pattern were measured for each group. Contact pressure was lowest in SRA-m followed by SRA-s. SRA-ama showed highest contact pressure of all single-row treatment groups (P < 0.05). DRA-amam presented the highest overall contact pressure (P < 0.05), whereas DRA-sm exerted contact pressure equal to that of SRA-ama. Both double-row techniques showed the most expanded pressure pattern. Average contact pressures for the more complex single- and double-row techniques utilizing arthroscopic Mason-Allen stitches were greater than were those of the repair techniques utilizing simple and horizontal mattress stitches. However, the contact pattern between the anchors could be increased by using the double-row technique, resulting in more footprint coverage compared to patterns utilizing the single-row techniques. These results support the use of the more complex arthroscopic Mason-Allen stitches and may improve the environment for healing of the repaired rotator cuff tendon. PMID- 19305973 TI - Computer-assisted revision of total knee arthroplasty. AB - Revision total knee prosthesis still remains a difficult procedure. Particularly, challenging is the restoration of the joint line to a normal position and the attainment of correct lower limb alignment and healthy bone support for the implants. Computer assistance improves accuracy during the implantation of primary total knees. The goal of this study was to evaluate the usefulness of computer assisted surgery (CAS) in total knee prosthesis revision. We revised 15 NKII total knee arthroplasties with the Navitrack system and compared the mechanical alignment and the joint line level on pre- and postoperative radiographs. After revision, the joint line position was restored. The knee with the revision prosthesis was aligned in the frontal plan with implants fixed perpendicularly to the mechanical axis. It is possible to correctly revise knee prosthesis with computer assistance only and without having to use a conventional ancillary. We had a permanent control of the joint line position and have performed the revision surgery following the same steps as in a primary implantation. PMID- 19305974 TI - Posterior stabilized TKA reduce patellofemoral contact pressure compared with cruciate retaining TKA in vitro. AB - Increased patellofemoral contact pressure was described after total knee arthroplasty (TKA). Aim of this in vitro study was to compare the influence of a posterior stabilized (PS) design in comparison to a cruciate retaining (CR) design on patellofemoral contact pressure. Patellofemoral area contact pressure, peak contact pressure and the centre of pressure motion were determined in eight fresh frozen human cadaveric specimens using a Tekscan sensor (K-Scan 4000). A robotic knee simulator was used simulating an isokinetic knee extension cycle from 120 degrees of flexion to full extension. All knees were tested in a first test cycle after implantation of a CR design and in a second test cycle after replacement by a PS design, both using a 11 mm PE inlay (Genesis II, Smith & Nephew, Memphis, TN, USA). The patella remained unresurfaced. A paired sampled t test to compare mean values (significance, P < or = 0.05) was used for statistical analysis. After implantation of the PS design, average patellofemoral area contact pressure was significantly lower (P < or = 0.006) compared with the CR design (PS: 3.58 +/- 1.25 MPa; CR: 4.31 +/- 1.40 MPa). Accordingly, average patellofemoral peak contact pressure decreased significantly (P < or = 0.02) with the PS design (6.12 +/- 2.37 MPa) in comparison with the CR design (7.17 +/- 2.41 MPa). On average, the centre of pressure motion was more physiological with the PS design compared to the CR design over the complete extension cycle. However, this was not significant. In conclusion, the data suggest less patellofemoral contact pressure of a posterior stabilized TKA design in comparison to a cruciate retaining design. PMID- 19305975 TI - Back pain and MRI changes in the thoraco-lumbar spine of top athletes in four different sports: a 15-year follow-up study. AB - A total 71 male athletes (weight lifters, wrestlers, orienteers, and ice-hockey players) and 21 non-athletes were randomly selected, for a baseline MRI study. After 15 years all the participants at baseline were invited to take part in a follow-up examination, including a questionnaire on back pain and a follow-up MRI examination. Thirty-two athletes and all non-athletes had disc height reduction at one or several disc levels. Disc degeneration was found in more than 90% of the athletes and deterioration had occurred in 88% of the athletes, with the highest frequency in weight lifters and ice-hockey players. 78% of the athletes and 38% of the non-athletes reported previous or present history of back pain at baseline and 71 and 75%, respectively at follow-up. There was no statistically significant correlation between back pain and MRI changes. In conclusion, athletes in sports with severe or moderate demands on the back run a high risk of developing disc degeneration and other abnormalities of the spine on MRI and they report high frequency of back pain. The study confirmed our hypothesis, i.e. that most of the spinal abnormalities in athletes seem to occur during the growth spurt, since the majority of the abnormalities demonstrated at follow-up MRI after the sports career were present already at baseline. The abnormalities found at young age deteriorated to a varying degree during the 15-year follow-up, probably due to a combination of continued high load sporting activities and normal ageing. Preventive measures should be considered to avoid the development of these injuries in young athletes. PMID- 19305976 TI - A novel implantation technique for engineered osteo-chondral grafts. AB - We present a novel method to support precise insertion of engineered osteochondral grafts by pulling from the bone layer, thereby minimizing iatrogenic damage associated with direct manipulation of the cartilage layer. Grafts were generated by culturing human expanded chondrocytes on Hyaff-11 meshes, sutured to Tutobone spongiosa cylinders. Through the bone layer, shaped to imitate the surface-contours of the talar dome, two sutures were applied: the first for anterograde implantation, to pull the graft into the defect, and the second for retrograde correction, in case of a too deep insertion. All grafts could be correctly positioned into osteochondral lesions created in cadaveric ankle joints with good fit to the surrounding cartilage. Implants withstood short term dynamic stability tests applied to the ankle joint, without delamination or macroscopic damage. The developed technique, by allowing precise and stable positioning of osteochondral grafts without iatrogenic cartilage damage, is essential for the implantation of engineered tissues, where the cartilage layer is not fully mechanically developed, and could be considered also for conventional autologous osteochondral transplantation. PMID- 19305977 TI - Histological and biomechanical studies of inter-strand healing in four-strand autograft anterior cruciate ligament reconstruction in a rabbit model. AB - In order to observe the remodeling process of the inter-strand of the four-strand graft used for anterior cruciate ligament (ACL) reconstruction, and to find the histological and biomechanical influences on this process by braiding the four strand graft, we conducted this study in a rabbit model. The rabbit's ACLs were reconstructed with bilateral four-strand semitendinous tendons which are in parallel or braided shape. The specimens of the two groups were collected at 3, 6, 12, 26 and 52 weeks after the operation. Gross and histological observations were done and the biomechanical properties of the specimens of the 26 and 52 weeks were compared. The result showed that in regular group, at 3 weeks, the strands which were in necrosis status were still separated with each other. At 6 weeks, the grafts were enveloped with hyperplastic synovium, some outer part of the gaps between strands were filled with synovium and fused together, while the inner parts were still separated. At 12 weeks, the graft was still in remodeling progress, but the remodeling degrees of each strand were different. Some strands had fused but some were still separated with "big" interval. At 26 and 52 weeks, some inter-strand gaps were hard to discriminate, some strands were connected with connective tissue and some separated ones still showed up. The braided ones manifested a similar but more synchronized remodeling process and showed a higher inter-strands fusing rate. The biomechanical test showed that the strength of the reconstructed ACL of the regular unbraided tendons was 36% of the contralateral normal ACL, while the braided ones reached 67%. The graft strengths of the specimen of the 26 and 52 weeks in the braided graft group were significantly higher than that of the unbraided group, while the stiffness of the 52-week specimen of the braided group was significantly higher than that of the unbraided group also. From this study, we concluded that as a graft for reconstruction of the ACL, the four-strand hamstring tendon needs to pass through the necrosis, revascularization, and ligamentation progress, but different strands were not in a synchronous process. The inter-strand gap may be completely fused, partial fused, fused but connected with connective tissue or still separated. By braiding the strands, the fusing percentage of the graft could be elevated and the biomechanical properties could be improved. PMID- 19305978 TI - Limited lactational transfer of acrylamide to rat offspring on maternal oral administration during the gestation and lactation periods. AB - To evaluate the developmental exposure effects of acrylamide (ACR) on the nervous and male reproductive systems, pregnant Sprague-Dawley rats were given ACR at 0, 25, 50 or 100 ppm in the drinking water from gestational day 6 to postnatal day (PND) 21 and histopathological assessment was performed at PND 21. Exposure levels in offspring were examined by measurement of free ACR and hemoglobin (Hb) ACR adducts on PND 14, and compared with maternal levels on PND 21. Additionally, a group of offspring that received ACR at 50 mg/kg by intraperitoneal injections directly three times a week from PND 2 to 21 was subjected to analysis for comparison with maternal exposure groups. Although maternal neurotoxicity was evident at 100 ppm, no changes suggestive of neurotoxicity or testicular toxicity were observed in their offspring except for growth retardation evident as lowered body weights. In contrast, offspring given ACR intraperitoneally exhibited obvious neurotoxicity, but not testicular damage. Free ACR in serum and milk was detected in neither dams nor their offspring. The level of ACR-Hb adducts in offspring was one tenth or less than that in dams. In summary, although preweaning rats have susceptibility to ACR-induced neurotoxicity, the internal level of ACR in offspring exposed through maternal oral administration is insufficient to induce neurotoxicity and testicular toxicity due to limited lactational transfer. PMID- 19305979 TI - A further plea for rigorous science and explicit disclosure of potential conflicts of interest. PMID- 19305980 TI - Determination of alkylphenols and alkylphenol ethoxylates in sewage sludge: effect of sample pre-treatment. AB - A complete characterization of sewage sludge collected from five biological waste water treatment plants was done to determine physico-chemical parameters, heavy metals and alkylphenols, making special emphasis on sampling, homogenization, and sample pre-treatment. Ultrasonic extraction followed by gas chromatrography coupled with mass spectrometry was used to evaluate the effect of sample pre treatment (untreated sample, freeze-drying, drying at 40 degrees C or drying at 100 degrees C) on the concentration of octylphenol (OP), nonylphenol (NP) and nonylphenol ethoxylates (NP1EO, NP2EO). Untreated samples and samples dried at 100 degrees C gave concentration levels up to 62% and 89% lower, respectively, than freeze-dried samples. In 50% of cases, freeze-dried samples led to significantly higher concentrations than those obtained by drying at 40 degrees C. Thus, freeze-drying is the recommended sample pre-treatment to prevent possible losses of OP, NP, and NP1EO. Using this methodology, concentrations detected were from 3.2 to 199 mg kg(-1) being NP followed by NP1EO found in highest concentration. The total concentration of NP and NP1EO exceeded the limit of 50 mg kg(-1) proposed by the draft European directive on sewage sludge in three out of five samples studied. Contrarily, heavy metals were below the legislated values. PMID- 19305981 TI - Gold nanoparticle dimer plasmonics: finite element method calculations of the electromagnetic enhancement to surface-enhanced Raman spectroscopy. AB - Finite element method calculations were carried out to determine extinction spectra and the electromagnetic (EM) contributions to surface-enhanced Raman spectroscopy (SERS) for 90-nm Au nanoparticle dimers modeled after experimental nanotags. The calculations revealed that the EM properties depend significantly on the junction region, specifically the distance between the nanoparticles for spacings of less than 1 nm. For extinction spectra, spacings below 1 nm lead to maxima that are strongly red-shifted from the 600-nm plasmon maximum associated with an isolated nanoparticle. This result agrees qualitatively well with experimental transmission electron microscopy images and localized surface plasmon resonance spectra that are also presented. The calculations further revealed that spacings below 0.5 nm, and especially a slight fusing of the nanoparticles to give tiny crevices, leads to EM enhancements of 10(10) or greater. Assuming a uniform coating of SERS molecules around both nanoparticles, we determined that regardless of the separation, the highest EM fields always dominate the SERS signal. In addition, we determined that for small separations less than 3% of the molecules always contribute to greater than 90% of the signal. PMID- 19305983 TI - Sound enhances touch perception. AB - Certain sounds, such as fingernails screeching down a chalkboard, have a strong association with somatosensory percepts. In order to assess the influences of audition on somatosensory perception, three experiments measured how task irrelevant auditory stimuli alter detection rates for near-threshold somatosensory stimuli. In Experiment 1, we showed that a simultaneous auditory stimulus increases sensitivity, but not response biases, to the detection of an electrical cutaneous stimulus delivered to the hand. Experiment 2 demonstrated that this enhancement of somatosensory perception is spatially specific--only monaural sounds on the same side increased detection. Experiment 3 revealed that the effects of audition on touch are also frequency dependent--only sounds with the same frequency as the vibrotactile frequency enhanced tactile detection. These results indicate that auditory information influences touch perception in highly systematic ways and suggest that similar coding mechanisms may underlie the processing of information from these different sensory modalities. PMID- 19305984 TI - The effect of altered gravity states on the perception of orientation. AB - We measured the effect of the orientation of the visual background on the perceptual upright (PU) under different levels of gravity. Brief periods of micro and hypergravity conditions were created using two series of parabolic flights. Control measures were taken in the laboratory under normal gravity with subjects upright, right side down and supine. Participants viewed a polarized, natural scene presented at various orientations on a laptop viewed through a hood which occluded all other visual cues. Superimposed on the screen was a character the identity of which depended on its orientation. The orientations at which the character was maximally ambiguous were measured and the perceptual upright was defined as half way between these orientations. The visual background affected the orientation of the PU less when in microgravity than when upright in normal gravity and more when supine than when upright in normal gravity. A weighted vector sum model was used to quantify the relative influence of the orientations of gravity, vision and the body in determining the perceptual upright. PMID- 19305985 TI - Individualised dosing of amikacin in neonates: a pharmacokinetic/pharmacodynamic analysis. AB - PURPOSE: To examine the pharmacokinetics of amikacin and its pharmacokinetic pharmacodynamic (PKPD) relationship in neonates. To develop an alternative dosing strategy for amikacin in neonates. METHODS: A population PKPD analysis was performed using data collected from 80 neonates with gestational ages from 24 to 41 weeks. The final pharmacokinetic model analysed 358 amikacin concentrations. All neonates were > 72 hours postnatal age. Simulations were performed to develop a new dosing strategy. RESULTS: The final covariate model was clearance = 0.23 x (current weight/2)(0.691) x (postmenstrual age/40)(3.23) and volume of distribution = 0.957 x (current weight/2)(0.89). Following the logistic regression analysis of treatment failure, new amikacin target concentrations were estimated and used in development of an alternative dosing strategy. CONCLUSION: Simulation of a new dosing regimen yielded the following recommendations: 15 mg/kg at 36-h intervals, 14 mg/kg at 24-h intervals and 15 mg/kg at 24-h intervals for neonates < or = 28 weeks, 29-36 weeks and > or = 37 weeks postmenstrual age respectively. PMID- 19305986 TI - The venous hinge--an objective sign for the diagnosis and follow-up of treatment in patients with intracranial hypotension syndrome. AB - INTRODUCTION: In patients with Intracranial Hypotension Syndrome (IHS), we observed reduction of the angle between vein of the Galen (VOG) and internal cerebral vein (ICV), which returns to the baseline after treatment. We coin the term "venous hinge" to describe this dynamic process and discuss its importance in IHS. METHODS: A midsagittal T1W image showing both VOG and ICV in the same plane was retrospectively analyzed by three different neuroradiologists in 17 patients with IHS. The angle between the lines drawn along the main axis of VOG and ICV (venous hinge angle (VHA)) was measured and documented. This angle measured from the magnetic resonance imaging (MRI) of the 50 normal controls was also recorded. Paired t tests were used to compare the VHA between male and female controls and between patients of IHS and normal controls. Sensitivity, specificity, positive (PPV) and negative predictive values (NPV) were calculated. The VHA was also calculated after treatment of these patients and paired t test was done to look for significant change in the VHA after treatment. RESULTS: The mean VHA formed by the veins in the IHS and control groups were 65 degrees (35-98 degrees) and 91 degrees (76-124 degrees) respectively (P < 0.0001). At a VHA of 79 degrees, the sensitivity, specificity, PPV, and NPV for diagnosis of IHS were 88.24%, 92%, 78.95%, and 95.83% respectively. In ten patients, follow-up MRI demonstrated normalization of the collapsing angle following appropriate treatment (p = 0.003). CONCLUSION: We report a previously undescribed imaging finding in patients with IHS. Recognition of this sign may aid in the diagnosis of IHS. PMID- 19305987 TI - Prenatal US diagnosis of congenital imperforate hymen. PMID- 19305988 TI - Assessment of the nature and age of subdural collections in nonaccidental head injury with CT and MRI. PMID- 19305989 TI - Beta amyloid peptide: from different aggregation forms to the activation of different biochemical pathways. AB - Amyloid beta peptide (Abeta) is the major component of amyloid plaques in the brain of individuals affected by Alzheimer's disease (AD). The formation of the plaques is due to an overproduction of Abeta by APP processing, its precursor, and to its ability to convert under specific conditions from its soluble form into highly ordered fibrillar aggregates. Although neuronal degeneration occurs near the amyloid plaques, some studies have suggested that intermediates such as protofibrils or simple oligomers are also involved in AD pathogenesis and even appear to be the more dangerous species in the onset of the pathology. Further, toxic properties of aggregates of different size have been investigated and the obtained results support the hypothesis that different aggregate sizes can induce different degeneration pathways. In the present review some of the knowledge about the biochemical routes of Abeta processing and production and the relationship among Abeta and oxidative stress, metal homeostasis, inflammatory process, and cell death are summarized. Moreover, current strategies addressing both fibrillogenesis process and different Abeta altered biochemical pathways utilized for therapies are described. PMID- 19305990 TI - Reminiscences of work with Alex Hope: the movement of water and ions in giant algal cells, 1963-1967. AB - This article, from the Tribute to Alex Hope Symposium at the 2008 Australian Society for Biophysics meeting, represents reminiscences of PhD studies done under my former supervisor, Professor Alex Hope. The studies demonstrated and quantified electroosmosis in giant algal cells of Chara and isolated segments of cell wall by measuring instantaneous current-induced volume flows. The studies also uncovered and modelled an unstirred-layer transport number effect that gave rise to an additional transiently increasing current-induced volume flow component, which could be mistaken for electroosmosis. In addition, action potential induced volume flows and pressure changes were measured in these cells and successfully modelled. An appreciation of the influence of Alex Hope and his laboratory environment, together with some of the further studies that resulted from this work, is also mentioned. PMID- 19305991 TI - Structure of the pore-helix of the hERG K(+) channel. AB - The hERG K(+) channel undergoes rapid inactivation that is mediated by 'collapse' of the selectivity filter, thereby preventing ion conduction. Previous studies have suggested that the pore-helix of hERG may be up to seven residues longer than that predicted by homology with channels with known crystal structures. In the present work, we determined structural features of a peptide from the pore loop region of hERG (residues 600-642) in both sodium dodecyl sulfate (SDS) and dodecyl phosphocholine (DPC) micelles using NMR spectroscopy. A complete structure calculation was done for the peptide in DPC, and the localization of residues inside the micelles were analysed by using a water-soluble paramagnetic reagent with both DPC and SDS micelles. The pore-helix in the hERG peptide was only two-four residues longer at the N-terminus, compared with the pore helices seen in the crystal structures of other K(+) channels, rather than the seven residues suggested from previous NMR studies. The helix in the peptide spanned the same residues in both micellar environments despite a difference in the localization inside the respective micelles. To determine if the extension of the length of the helix was affected by the hydrophobic environment in the two types of micelles, we compared NMR and X-ray crystallography results from a homologous peptide from the voltage gated potassium channel, KcsA. PMID- 19305992 TI - Mixed fermentation for natural product drug discovery. AB - Natural products continue to play a major role in drug discovery and development. However, chemical redundancy is an ongoing problem. Genomic studies indicate that certain groups of bacteria and fungi have dozens of secondary metabolite pathways that are not expressed under standard laboratory growth conditions. One approach to more fully access the metabolic potential of cultivatable microbes is mixed fermentation, where the presence of neighboring microbes may induce secondary metabolite synthesis. Research to date indicates that mixed fermentation can result in increased antibiotic activity in crude extracts, increased yields of previously described metabolites, increased yields of previously undetected metabolites, analogues of known metabolites resulting from combined pathways and, importantly, induction of previously unexpressed pathways for bioactive constituents. PMID- 19305993 TI - Bioaugmentation of UASB reactors with immobilized Sulfurospirillum barnesii for simultaneous selenate and nitrate removal. AB - Whole-cell immobilization of selenate-respiring Sulfurospirillum barnesii in polyacrylamide gels was investigated to allow the treatment of selenate contaminated (790 microg Se x L(-1)) synthetic wastewater with a high molar excess of nitrate (1,500 times) and sulfate (200 times). Gel-immobilized S. barnesii cells were used to inoculate a mesophilic (30 degrees C) bioreactor fed with lactate as electron donor at an organic loading rate of 5 g chemical oxygen demand (COD) x L(-1) day(-1). Selenate was reduced efficiently (>97%) in the nitrate and sulfate fed bioreactor, and a minimal effluent concentration of 39 microg Se x L(-1) was obtained. Scanning electron microscopy with energy dispersive X-ray (SEM-EDX) analysis revealed spherical bioprecipitates of 0.05). The data indicated a statistically significant reduction of ulcer size (p < 0.001), pain severity (p < 0.05), and erythema and exudation level (p < 0.001). Oral Health Impact Profile improved significantly in the treatment group (p < 0.001). Patient overall assessment of their treatment improved after applying paste containing myrtle (p < 0.05). No side effects were reported. This study has shown myrtle to be effective in decreasing the size of ulcers, pain severity and the level of erythema and exudation, and improving the quality of life in patients who suffer from RAS. PMID- 19306025 TI - The evaluation of a novel method comparing quantitative light-induced fluorescence (QLF) with spectrophotometry to assess staining and bleaching of teeth. AB - This study reports the development and evaluation of a novel method using quantitative light-induced fluorescence (QLF), which enables its use for quantifying and assessing whole tooth surface staining and tooth whitening. The method was compared with a spectrophotometer to assess reliability. Two experimental phases, intrinsic stain formation and tooth whitening, were conducted in vitro on 16 extracted bovine teeth. Intrinsic stains were developed via access through lingual surfaces and root canals of these teeth using tea solution (2 g/100 ml, Marks and Spencer Extra Strong Tea, Marks and Spencer, London, UK) for 6 days. Stains were removed using 33% hydrogen peroxide (VWR Prolab, Leicestershire, UK) in cycles over 150 min. Stain development/whitening was monitored with QLF (Inspektor Research systems, Amsterdam, Netherlands) and spectrophotometry (Easy shade, Vita Zahnfabrik, Bad Sackingen, Germany). Parameters Delta F for QLF and Delta E for the spectrophotometer were obtained. The progression of stain intensity and removal observed by the methods were tested for correlation using Pearson's correlation coefficient. Intra-examiner reliability for each method was tested. QLF showed a high correlation with spectrophotometry for detecting and monitoring intrinsic tooth stain progression (Pearson coefficient r was -0.987 with correlation significant p < 0.0001). For stain removal, the Pearson coefficient (r) between both methods was -0.906 with no significance p = 0.094. The use of an external reference material in combination with the inner patch QLF analysis technique had the ability to detect and measure whole tooth surface staining and its removal longitudinally. The reliability of the method shows a potential clinical application. PMID- 19306026 TI - Effective potentials of dissipative hard spheres in granular matter. AB - We present an experimental study of the spatial correlations of a quasi-two dimensional dissipative gas kept in a non-static steady state via vertical shaking. From high temporal resolution images we obtain the Pair Distribution Function (PDF) for granular species with different restitution coefficients. Effective potentials for the interparticle interaction are extracted using the Ornstein-Zernike equation with the Percus-Yevick closure. From both the PDFs and the corresponding effective potentials, we find a clear increase of the spatial correlation at contact with the decreasing values of the restitution coefficient. PMID- 19306027 TI - First REP-LECOTOX Workshop-Ecotoxicogenomics: The challenge of integrating genomics/proteomics/metabolomics into aquatic and terrestrial ecotoxicology. PMID- 19306028 TI - Atmospheric fall-out of metals around the Murano glass-making district (Venice, Italy). AB - BACKGROUND, AIM AND SCOPE: Murano's glass-makers have held a monopoly on quality glass-making for centuries known all over the world. Artistic glass manufacture entails exposure to complex mixtures of pollutants, including metals. A few studies have reported high levels of trace elements in marine waters, sediments and mussels around Murano and shown that emissions from Murano glass-making workshops significantly influence air quality in the Venice area. Nevertheless, to date, there is very little information on atmospheric concentrations and virtually none on atmospheric deposition fluxes of trace elements around the island. This study presents data on the distribution of trace elements in the air and atmospheric depositions around Murano, based on a 2-year sampling period. MATERIALS AND METHODS: Airborne PM10 particulate matter was collected daily in the period December 2001-June 2003 (254 air samples), and atmospheric depositions were collected every 19 +/- 6 days, in the period August 2001-July 2003 (38 samples) on the roof of the Experimental Glass Laboratory on the island of Murano. All samples were analysed by ICP-MS for As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Sb, Se and Zn contents. RESULTS: Atmospheric concentrations of As, Cd, Cr, Ni, Pb, Se and Zn were 1-2 orders of magnitude greater on the island of Murano than in the nearest urban areas (Venice and Mestre), with values higher than those reported for European industrial sites. The atmospheric deposition fluxes of all elements were also higher in Murano than those detected by other studies in Venice and Mestre and in the Porto Marghera industrial zone, with As, Cd, Sb and Se 1-2 orders of magnitude higher. DISCUSSION: In order to study the spatial variability of the atmospheric fall-out, data from other sampling stations belonging to Venice atmospheric deposition monitoring networks were used. A decreasing gradient from Murano to the closer leeward stations was observed for As, Cd and Se. Pb and Zn deposition fluxes in Venice were also comparable to those observed in the Porto Marghera industrial zone, thus, indicating a similar or even higher level of contamination. Principal component analysis confirmed significant contamination from the glassworks. CONCLUSIONS: As shown by our study, atmospheric loadings of metals around Murano are significant. These observations confirm that emissions from Murano also significantly influence atmospheric deposition in the Venice area. Specifically, the mean daily Cd flux in the most affected area, which includes the whole of the historic city centre of Venice, is approximately 18 microg m(-2) day(-1), i.e. more than 65 times higher than the Dutch limit and more than three times higher than that of Germany. RECOMMENDATIONS AND PERSPECTIVES: On the basis of our data, there is a clear-cut need for remedial action in the Lagoon of Venice. Monitoring is indispensable, so that the efficacy of remedial measures can be evaluated and appropriate information about risks for human health and well-being can be made available. The atmospheric compartment must also be considered by Italian and European law which, until now, has not yet established any standard for atmospheric deposition. PMID- 19306029 TI - Effect of rotator cuff dysfunction on the initial mechanical stability of cementless glenoid components. AB - The functional outcome of shoulder replacement is related to the condition of the rotator cuff. Rotator cuff disease is a common problem in candidates for total shoulder arthroplasty; this study relates the functional status of the rotator cuff to the initial stability of a cementless glenoid implant. A 3D finite element model of a complete scapula was used to quantify the effect of a dysfunctional rotator cuff in terms of bone-implant interface micromotions when the implant is physiologically loaded shortly after surgery. Four rotator cuff conditions (from fully intact to progressively ruptured rotator cuff tendons) as well as two bone qualities were simulated in a model. Micromotions were significantly larger in the worst modeled cuff dysfunction (i.e. the supraspinatus and infraspinatus tendons were fully dysfunctional). Micromotions were also significantly different between conditions with healthy and poor bone quality. The implant's initial stability was hardly influenced by a dysfunctional supraspinatus alone. However, when the infraspinatus was also affected, the glenohumeral joint force was displaced to the component's rim resulting in larger micromotions and instability of the implant. PMID- 19306030 TI - Cardiac anisotropy in boundary-element models for the electrocardiogram. AB - The boundary-element method (BEM) is widely used for electrocardiogram (ECG) simulation. Its major disadvantage is its perceived inability to deal with the anisotropic electric conductivity of the myocardial interstitium, which led researchers to represent only intracellular anisotropy or neglect anisotropy altogether. We computed ECGs with a BEM model based on dipole sources that accounted for a "compound" anisotropy ratio. The ECGs were compared with those computed by a finite-difference model, in which intracellular and interstitial anisotropy could be represented without compromise. For a given set of conductivities, we always found a compound anisotropy value that led to acceptable differences between BEM and finite-difference results. In contrast, a fully isotropic model produced unacceptably large differences. A model that accounted only for intracellular anisotropy showed intermediate performance. We conclude that using a compound anisotropy ratio allows BEM-based ECG models to more accurately represent both anisotropies. PMID- 19306031 TI - A sideways glance. Alcoholic breakdown of barriers: how ethanol can initiate a landslide towards disease. PMID- 19306033 TI - Fibre in beverages can enhance perceived satiety. AB - BACKGROUND: A high intake of dietary fibre has been suggested to support the regulation of energy intake and satiety, which could contribute favourably to the increasing obesity problem. AIM OF THE STUDY: To investigate the effects of three fibres differing in chemical and physical properties on perceived satiety and hunger-related attributes. METHODS: A total of 19 healthy volunteers, age 18-30, mean BMI 23.2 kg/m(2) participated in the study. Measurement of food and satiety related perceptions with ten attributes was performed by using 10-unit graphic intensity scales during a 120 min period after the ingestion the sample. The attributes evaluated were satiety, hunger (unipolar and bipolar scale), appetite, fullness, desire to eat something/sweet/savoury/the sample food and thirst. The sample foods used were a beverage without fibre, a guar gum beverage, a wheat bran beverage, an oat beta-glucan beverage and wheat bread was used as the control. The fibre content of the samples was 0 g (beverage without fibre), 2.4 g (wheat bread), 7.8 g (guar gum) or 10.5 g (wheat bran and oat beta-glucan beverage) per 400 g/1,000 kJ portion. RESULTS: The area under curve (AUC) for perceived satiety was higher (169 vs. 83 cm min; t test P = 0.026) and the desire to eat was lower (AUC -179 vs. -83 cm min; t test P = 0.008) for the guar gum beverage as compared to the beverage without fibre. Also the beverage with oat beta-glucan increased fullness and showed a trend of increasing perceived satiety and decreasing the desire to eat more than the beverage without fibre. CONCLUSIONS: Our results support the idea that dietary fibre in beverages can enhance their perceived satiety and decrease the desire to eat more than a beverage without fibre. PMID- 19306034 TI - Quinine-responsive muscle cramps in X-linked bulbospinal muscular atrophy Kennedy. PMID- 19306035 TI - Hypermetabolism in ALS patients: an early and persistent phenomenon. AB - The malnutrition common among patients with ALS can be attributed in some cases to increased resting energy expenditure (REE). However, the origins and evolution of this hypermetabolism have yet to be fully elucidated. The aim of the present study was to monitor REE over time in patients with ALS and to identify factors that may explain any variation observed. ALS patients underwent nutritional, neurological and respiratory assessment every 6 months for 2 years (or until they died or became physically incapable of being examined). Sixty-one patients were studied. At inclusion, 47.5% exhibited hypermetabolism, with a mean measured REE (mREE) 19.7 +/- 6.4% higher than the mean calculated REE (cREE) (P < 0.0001). The hypermetabolism persisted when mREE was normalized for fat free mass (FFM): 35.1 +/- 4.2 versus 32.3 +/- 4.7 kcal/kg day(-1) (P = 0.02) in hypermetabolic and normometabolic patients, respectively. In univariate analysis, mREE was negatively correlated with age and positively correlated with BMI, FFM, energy and protein intakes, and albumin level. No correlation was found with neurological scores, disease characteristics, respiratory function and survival. Multivariate analysis revealed no significant factors. Only 10 of 45 patients in whom REE was measured at least twice changed their metabolic status. Neither mREE nor mREE/cREE varied significantly over time, despite deteriorating neurological, nutritional and respiratory parameters (P < 0.0001), and an increase in mREE/FFM (P = 0.01). This study confirms that about 50% of ALS patients are hypermetabolic, and 80% show no change in metabolic status over time. Thus, metabolic status (a clinically useful indicator of the need for nutritional support) can be determined early in the evolution of the disease. The origin of hypermetabolism in this context remains unknown, but growing evidence points to mitochondria as having an important role. PMID- 19306036 TI - The role of endotoxin in critical illness myopathy and polyneuropathy. PMID- 19306037 TI - Sensory "pseudoperipheral" migraine aura. PMID- 19306038 TI - A double blind placebo RCT to investigate the effects of serotonergic modulation on brain excitability and motor recovery in stroke patients. AB - Motor excitability is increased in both hemispheres in stroke patients during motor recovery. Pharmacologically controlled changes of cortical excitability might be beneficial for synaptic plasticity and therefore facilitate functional recovery after a brain lesion. In particular, it has been suggested that antidepressant drugs can modulate motor excitability. Several recent reports suggest the possibility of monitoring pharmacological effects on brain excitability through transcranial magnetic stimulation (TMS). The aim of this study was to investigate motor area excitability in patients with stroke after oral administration of citalopram. We conducted a prospective randomised placebo controlled study. Twenty patients with unilateral stroke were included in the study: ten patients treated by antidepressive drug and ten patients with placebo. A selective serotonergic drug (citalopram) or a placebo was administered using a mean dosage of 10 mg/day in combination with physiotherapy. Motor cortex excitability was studied by single and paired transcranial magnetic stimulation. TMS recording was tested before (T1) and 1 month after (T2) beginning drug treatment. Patients treated by the serotonergic drug, compared to patients that received a placebo, showed a significant improvement in neurological status as measured by NIHSS and a decrease of motor excitability over the unaffected hemisphere, while no differences were observed over the affected hemisphere. Our findings suggest that treatment with serotonergic drugs can bring about a significant decrease of the motor cortex excitability in stroke patients with effects on both the affected and unaffected hemispheres associated with a better motor recovery. PMID- 19306039 TI - Prednisone 10 days on/10 days off in patients with Duchenne muscular dystrophy. AB - Corticosteroids are effective in improving motor function in Duchenne muscular dystrophy (DMD) patients within 6 months-2 years of treatment initiation, but there is as yet no consensus on which treatment scheme is the best. We retrospectively analyzed data of 35 DMD patients who were treated with prednisone 0.75 mg/kg per day intermittently 10 days on/10 days off. Prednisone was started during the ambulant phase at age 3.5-9.7 years (median 6.5 years). The median period of treatment was 27 months (range 3-123 months). The median age at which ambulation was lost was 10.8 years (mean 10.9 years; 95% confidence interval 10.0 11.8 years). Nine patients (26%) had excessive weight gain. Eight boys (21%) had a bone fracture, which was when four of these eight children lost the ability to walk. Treatment was stopped in two obese patients, two hyperactive boys and one patient following a fracture. Our data suggest that prednisone 10 on/10 off has relatively few side effects and extends the ambulant phase by 1 year compared to historical controls. PMID- 19306040 TI - Erwin Gustav Niessl von Mayendorf (1873-1943). PMID- 19306041 TI - Ocular fixation instabilities in motor neurone disease. A marker of frontal lobe dysfunction? AB - OBJECTIVE: Eye movements are classically felt to be spared in motor neurone disease (MND). Although a range of ocular motor disorders have been reported, no consistent pattern has been established. Disturbances of ocular fixation have been noted in MND; however, fixation has not yet been formally examined. With the recent characterization of ocular fixation using saccadic intrusion amplitude and fixation periods, we performed a cross-sectional study to examine for abnormalities of ocular fixation in non-dementing patients with MND. METHODS: A total of 44 patients and 45 controls were recruited. Fixation was examined using infra-red oculography and all subjects then underwent a neuropsychological evaluation. RESULTS: Saccadic intrusion amplitude was found to be greater in patients compared to controls and in particular, spinal-onset patients. Saccadic intrusion amplitude in patients correlated with neuropsychological measures sensitive to lesions of the frontal lobes. CONCLUSIONS: This is the first study to identify abnormalities of fixation in MND and these results indicate that ocular fixation instabilities may be a marker of the sub-clinical frontal lobe dysfunction in MND. A longitudinal study to examine if saccadic intrusion amplitude deteriorates with time would be of interest as this could provide a quantifiable objective marker of disease progression. PMID- 19306042 TI - Characterization of a hemolysin gene ytjA from Bacillus subtilis. AB - Bacillus subtilis is popularly used as a probiotic in many fields. Although recent research has found that some secondary metabolites of B. subtilis could cause hemolysis, the hemolytic mechanism in B. subtilis is still unclear. In this paper, the hemolysis-associated gene ytjA was cloned and expressed in Escherichia coli, and the hemolytic activity of the expressed soluble protein was indicated by the presence of clear hemolytic zones on sheep blood agar plates. In addition, reverse transcriptase-polymerase chain reaction analysis confirmed that the ytjA gene was transcribed in B. subtilis. These results suggest that ytjA is one of the hemolysin genes responsible for hemolysis in B. subtilis. PMID- 19306043 TI - A genome-wide survey on basic helix-loop-helix transcription factors in rat and mouse. AB - The basic helix-loop-helix (bHLH) proteins play essential roles in a wide range of developmental processes in higher organisms. bHLH family members have been identified in over 20 organisms, including nematode, fruit fly, and human. Our study identified 114 rat and 14 additional mouse bHLH members in rat and mouse genomes, respectively. Phylogenetic analyses revealed that both rat and mouse had 49, 26, 15, 4, 12, and 4 bHLH members in groups A, B, C, D, E, and F, respectively. Only the rat Mxi1 gene has two copies in the genome. All other rat bHLH genes and all mouse bHLH genes are single-copy genes. The chromosomal distribution pattern of mouse, rat, and human bHLH genes suggests the emergence of some bHLH genes through gene duplication, which probably happened at least before the divergence of vertebrates from invertebrates. The present study provides useful information for future studies using rat as a model animal for mammalian development. PMID- 19306044 TI - Replication of long-bone length QTL in the F9-F10 LG,SM advanced intercross. AB - Quantitative trait locus (QTL) mapping techniques are frequently used to identify genomic regions associated with variation in phenotypes of interest. However, the F(2) intercross and congenic strain populations usually employed have limited genetic resolution resulting in relatively large confidence intervals that greatly inhibit functional confirmation of statistical results. Here we use the increased resolution of the combined F(9) and F(10) generations (n = 1455) of the LG,SM advanced intercross to fine-map previously identified QTL associated with the lengths of the humerus, ulna, femur, and tibia. We detected 81 QTL affecting long-bone lengths. Of these, 49 were previously identified in the combined F(2) F(3) population of this intercross, while 32 represent novel contributors to trait variance. Pleiotropy analysis suggests that most QTL affect three to four long bones or serially homologous limb segments. We also identified 72 epistatic interactions involving 38 QTL and 88 novel regions. This analysis shows that using later generations of an advanced intercross greatly facilitates fine mapping of confidence intervals, resolving three F(2)-F(3) QTL into multiple linked loci and narrowing confidence intervals of other loci, as well as allowing identification of additional QTL. Further characterization of the biological bases of these QTL will help provide a better understanding of the genetics of small variations in long-bone length. PMID- 19306047 TI - Desmoids: a revelation in biology and treatment. AB - INTRODUCTION: Using a preoperative neoadjuvant chemoradiation protocol, followed by complete excision, we have achieved local control rates exceeding that found in most large series. METHODS: From October 1990 through May 2008, resectable desmoids were initially treated with a preoperative protocol using Adriamycin 30 mg x 3 days continuous intravenous infusion followed by 3,000 cGy of radiation (300 cGy fractions over 10 days). Resection was performed 4-6 weeks later. After 2001, all patients were initially offered Tamoxifen 120 mg/day and Celebrex 400 mg/day for 1 year. Patients who progressed on Tamox/Celeb were treated with protocol and those with stabilization or regression were observed. Patient demographics, tumor size, history of previous recurrences, and follow-up status were recorded prospectively. RESULTS: There were 40 females and 12 males with a mean follow-up of 45 months. Forty patients presented with primary tumors and eight presented with recurrent disease. Thirty-nine patients had surgical resection and 13 were observed. Thirty patients underwent the neoadjuvant protocol. Tamoxifen and Celebrex were used in 16 patients, 6 had stabilization in growth, 1 had a 50% reduction in the size of the tumor, there was 1 complete regression, and 8 progressed. Of the patients who had resectable disease Tamoxifen and Celebrex obviated surgery in 30%. Overall 13% (5) of patients developed a recurrence. There were three recurrences among the protocol group for a local control rate of 90%. CONCLUSION: Although our neoadjuvant protocol demonstrates the best results to date in eradication of disease, an initial conservative approach is reasonable to determine who would most benefit from surgery. PMID- 19306048 TI - Nerve allotransplantation as it pertains to composite tissue transplantation. AB - Nerve allografts provide a temporary scaffold for host nerve regeneration and allow for the repair of significant segmental nerve injuries. From rodent, large animal, and nonhuman primate studies, as well as clinical experience, nerve allografts, with the use of immunosuppression, have the capacity to provide equal regeneration and function to that of an autograft. In contrast to solid organ transplantation and composite tissue transfers, nerve allograft transplantation requires only temporary immunosuppression. Furthermore, nerve allograft rejection is difficult to assess, as the nerves are surgically buried and are without an immediate functional endpoint to monitor. In this article, we review what we know about peripheral nerve allograft transplantation from three decades of experience and apply our current understanding of nerve regeneration to the emerging field of composite tissue transplantation. PMID- 19306049 TI - Total distal radioulnar joint arthroplasty in adults with symptomatic Madelung's deformity. AB - Madelung's deformity is a well-known congenital anomaly of the wrist. A number of procedures have been described to correct the deformity and thus improve function at the wrist. Most of these procedures have to a large extent addressed the alignment and consequent function of the radiocarpal joint but not the persistent problem of painful stiffness at the distal radioulnar joint (DRUJ). The availability of a total DRUJ prosthesis appears to provide a solution to this problem. This article discusses our early experience with total DRUJ arthroplasty using the Scheker prosthesis for managing pain and decreased range of motion in three adult patients with symptomatic Madelung's deformity. PMID- 19306050 TI - Genetic testing before anticoagulation? A systematic review of pharmacogenetic dosing of warfarin. AB - BACKGROUND: Genotype-guided initial warfarin dosing may reduce over anticoagulation and serious bleeding compared to a one-dose-fits-all dosing method. OBJECTIVE: The objective of this review was to investigate the safety and efficacy of genotype-guided dosing of warfarin in reducing the occurrence of serious bleeding events and over-anticoagulation. DATA SOURCES: The authors searched PubMed, EMBASE and International Pharmaceutical Abstracts through January 23, 2009, without language restrictions. Selected articles were randomized trials comparing pharmacogenetic dosing of warfarin versus a "standard" dose control algorithm in adult patients taking warfarin for the first time. REVIEW METHODS: Two reviewers independently extracted data and assessed study quality using a validated instrument. The primary outcomes were major bleeding and time spent within the therapeutic range International Normalized Ratio (INR). Secondary outcomes included minor bleeding, thrombotic events and other measures of anticoagulation quality. RESULTS: Three of 2,014 studies (423 patients) met the inclusion and exclusion criteria. Differences in study quality, dosing algorithms, length of follow-up and outcome measures limited meta analysis. Summary estimates revealed no statistically significant difference in bleeding rates or time within the therapeutic range INR. The highest quality study found no significant difference in primary or secondary outcomes, although there was a trend towards more rapid achievement of a stable dose (14.1 vs. 19.6 days, p = 0.07) in the pharmocogenetic arm. CONCLUSIONS: We did not find sufficient evidence to support the use of pharmacogenetics to guide warfarin therapy. Additional clinical trials are needed to define the optimal approach to use warfarin pharmacogenetics in clinical practice. PMID- 19306051 TI - Continuous infusion of intraperitoneal bupivacaine after laparoscopic surgery: a randomized controlled trial--what about statistical power and analysis? PMID- 19306052 TI - The psychology of bariatric patient: what replaces obesity? A qualitative research with Brazilian women. AB - BACKGROUND: Obesity has serious implications on a woman's quality of life and body image. We propose a qualitative investigation aimed at understanding the postoperative significance of bariatric surgery for women suffering from morbid obesity and how these factors influence the outcomes, with an emphasis on body image and on the possible psychological complications that may jeopardize the operation's success. SUBJECTS AND METHODS: This study uses a clinical qualitative method, through a semidirected interview with open-ended questions in an intentional sample, closed by saturation, with seven women operated in a period of 1.5-3 years, following the definition of emergent categories and qualitative content analysis. RESULTS: Bariatric surgery is a procedure that brings about rapid physical, social, and emotional changes, and it is seen by patients as a possibility of being reinstated and accepted socially. The reencounter with the feminine body after surgery is experienced as a means of reinstatement but also with a feeling of defenselessness, which may lead to the development of phobic symptoms. Imbalance in family and conjugal relationships may be factors that discourage the continuation of the treatment. The patient sees the skin folds, flaccidity, and the scars as therapeutic failures, which can lead to a constant quest for plastic surgery. CONCLUSION: We observe the necessity of studies that allow the health team to identify those aspects of a patient's psychological makeup which would be expected to improve or worsen their prognosis and to provide the necessary preoperative and postoperative psychosocial interventions. PMID- 19306055 TI - ICAM-1 (Lys469Glu) and PECAM-1 (Leu125Val) polymorphisms in diffuse astrocytomas. AB - Cell adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1) and platelet-endothelial cell adhesion molecule-1 (PECAM-1) play an important role in glioma invasion and angiogenesis. The aim of this study was to investigate whether specific genetic polymorphisms of ICAM-1 and PECAM-1 could be associated with glioma development and progression. Single-nucleotide polymorphism in codon 469 of ICAM-1 and codon 125 of PECAM-1 were examined in 158 patients with astrocytomas and 162 controls using polymerase chain reaction and restriction enzyme analysis. The distribution of PECAM-1 polymorphic genotypes in astrocytomas did not show any significant difference. However, a specific ICAM-1 genotype (G/G, corresponding to Lys469Glu) exhibited higher frequency in grade II astrocytomas compared to controls, grade III, and grade IV astrocytomas; suggesting that this polymorphism could be involved in the development of grade II astrocytomas. PMID- 19306056 TI - Abnormal muscles that may affect axillary lymphadenectomy: surgical anatomy. AB - Purpose The present study aimed at summarizing and presenting the anomalous muscles that a surgeon might encounter during axillary lymphadenectomy (AL). Methods For this purpose, both the anatomical and surgical literature was reviewed and an anatomical study on 107 cadavers was carried out. Furthermore, based on the anatomical features of the anomalous muscles that came up during our study and taking into consideration the landmarks of the AL, we further analyzed the complications that may arise from each of these muscles, along with their preoperative and intraoperative recognition and management. Results The literature review revealed that there are three supernumerary muscles that may affect the AL, namely the Langer's axillary arch, the pectoralis quartus and the chondroepitrochlearis muscles, as well as the aplasia of the lower part of the pectoralis major muscle. Eight out of the 107 (7.48%) cadavers that we dissected had such an abnormal muscle in the axilla. Specifically, the axillary arch was found unilaterally in five cadavers (4.67%) and the pectoralis quartus muscle was present unilaterally in three cadavers (2.8%). One cadaver had both an axillary arch and a pectoralis quartus muscle in the right side. The abdominal and almost the whole sternocostal portion of the pectoralis major as well the pectoralis minor muscle were absent in one cadaver (0.93%). The chondroepitrochlearis muscle was not found in any of the cadavers that we dissected. Conclusions The present study offers the necessary preoperative knowledge for recognizing these muscles during AL, avoiding thus the complications that may arise from them. PMID- 19306057 TI - Axillary web syndrome after axillary dissection in breast cancer: a prospective study. AB - The axillary web syndrome is a self-limiting and frequently overlooked cause of significant morbidity in the early post-operative period after breast cancer axillary surgery, which is characterized by axillary pain that runs down the medial arm, limited shoulder range of motion affecting mainly shoulder abduction, and cords of subcutaneous tissue extending from axilla into the medial arm, made visible or palpable and painful by shoulder abduction. We evaluated the incidence of axillary web syndrome after axillary lymph-node dissection. Altogether 116 patients who underwent axillary lymph-node dissection were assessed for axillary web syndrome. Range of shoulder abduction, pain, and the subjective feeling of tightness along the arm were recorded. Axillary web syndrome was found in 56 patients, which means an incidence of 48.3% (CI95%: 38.9, 57.7). Axillary web syndrome is a significant self-limited cause of morbidity of axillary surgery ocurring not only in the early post-operative period. PMID- 19306058 TI - Aquaporin 1 (AQP1) expression is a novel characteristic feature of a particularly aggressive subgroup of basal-like breast carcinomas. AB - Aquaporin1 (AQP1) is a water channel protein that facilitates water flux across cell membranes. It is widely expressed in epithelial and endothelial cells in several tissues. AQP1 is also associated with angiogenesis, cell migration and metastasis in some human malignancies. In this study the immunohistochemical expression of AQP1 in 203 invasive breast carcinomas with long-term follow up was investigated. AQP1 expression was demonstrated in 11 tumours (5.4%) and showed highly significant correlation with high tumour grade, medullary-like histology, "triple-negativity", cytokeratin 14 and smooth muscle actin expression. In univariate analysis, AQP1 was significantly associated with poor prognosis. In multivariate analysis, AQP1 expression proved to be an independent prognostic marker if stratified by age, tumour size, lymph node status, histological grade, ER status and CMF therapy. Our results strongly suggest that AQP1 expression is a new characteristic feature of a particularly aggressive subgroup of basal-like breast carcinomas. PMID- 19306059 TI - Differentiation of benign and malignant breast lesions by mechanical imaging. AB - Mechanical imaging yields tissue elasticity map and provides quantitative characterization of a detected pathology. The changes in the surface stress patterns as a function of applied load provide information about the elastic composition and geometry of the underlying tissue structures. The objective of this study is the clinical evaluation of breast mechanical imager for breast lesion characterization and differentiation between benign and malignant lesions. The breast mechanical imager includes a probe with pressure sensor array, an electronic unit providing data acquisition from the pressure sensors and communication with a touch-screen laptop computer. We have developed an examination procedure and algorithms to provide assessment of breast lesion features such as hardness related parameters, mobility, and shape. A statistical Bayesian classifier was constructed to distinguish between benign and malignant lesions by utilizing all the listed features as the input. Clinical results for 179 cases, collected at four different clinical sites, have demonstrated that the breast mechanical imager provides a reliable image formation of breast tissue abnormalities and calculation of lesion features. Malignant breast lesions (histologically confirmed) demonstrated increased hardness and strain hardening as well as decreased mobility and longer boundary length in comparison with benign lesions. Statistical analysis of differentiation capability for 147 benign and 32 malignant lesions revealed an average sensitivity of 91.4% and specificity of 86.8% with a standard deviation of +/-6.1%. The area under the receiver operating characteristic curve characterizing benign and malignant lesion discrimination is 86.1% with the confidence interval ranging from 80.3 to 90.9%, with a significance level of P = 0.0001 (area = 50%). The multisite clinical study demonstrated the capability of mechanical imaging for characterization and differentiation of benign and malignant breast lesions. We hypothesize that the breast mechanical imager has the potential to be used as a cost effective device for cancer diagnostics that could reduce the benign biopsy rate, serve as an adjunct to mammography and to be utilized as a screening device for breast cancer detection. PMID- 19306060 TI - The State Health Authority Yardstick (SHAY). AB - State mental health authorities have a leadership role in implementing evidence based practices (EBPs), but few instruments are available to assess the impact of this role. We describe the development of the State Mental Health Authority Yardstick (SHAY), a behaviorally anchored instrument designed to assess state level facilitating conditions associated with successful implementation of EBPs in community mental health centers. The SHAY assesses the SMHA role in seven domains: Planning, Financing, Training, Leadership, Policies and Regulations, Quality Improvement, and Stakeholders. Preliminary evidence from the National Evidence-Based Practices Project partially supports the construct and criterion oriented validity of this instrument for rating state-level activities supporting or blocking the implementation of evidence-based practices. PMID- 19306061 TI - Phonological awareness predicts activation patterns for print and speech. AB - Using fMRI, we explored the relationship between phonological awareness (PA), a measure of metaphonological knowledge of the segmental structure of speech, and brain activation patterns during processing of print and speech in young readers from 6 to 10 years of age. Behavioral measures of PA were positively correlated with activation levels for print relative to speech tokens in superior temporal and occipito-temporal regions. Differences between print-elicited activation levels in superior temporal and inferior frontal sites were also correlated with PA measures with the direction of the correlation depending on stimulus type: positive for pronounceable pseudowords and negative for consonant strings. These results support and extend the many indications in the behavioral and neurocognitive literature that PA is a major component of skill in beginning readers and point to a developmental trajectory by which written language engages areas originally shaped by speech for learners on the path toward successful literacy acquisition. PMID- 19306062 TI - Glycosylation of aromatic amines I: Characterization of reaction products and kinetic scheme. AB - The reactions of aliphatic and aromatic amines with reducing sugars are important in both drug stability and synthesis. The formation of glycosylamines in solution, the first step in the Maillard reaction, does not typically cause browning but results in decreased potency and is hence significant from the aspect of drug instability. The purpose of this research was to present (1) unreported ionic equilibria of model reactant (kynurenine), (2) the analytical methods used to characterize and measure reaction products, (3) the kinetic scheme used to measure reaction rates and (4) relevant properties of various reducing sugars that impact the reaction rate in solution. The methods used to identify the reversible formation of two products from the reaction of kynurenine and monosaccharides included LC mass spectrometry, UV spectroscopy, and 1-D and 2 D (1)H-(1)H COSY NMR spectroscopy. Kinetics was studied using a stability indicating HPLC method. The results indicated the formation of alpha and beta glycosylamines by a pseudo first-order reversible reaction scheme in the pH range of 1-6. The forward reaction was a function of initial glucose concentration but not the reverse reaction. It was concluded that the reaction kinetics and equilibrium concentrations of the glycosylamines were pH-dependent and also a function of the acyclic content of the reacting glucose isomer. PMID- 19306063 TI - Disclosing HIV serostatus to family members: Effects on psychological and physiological health in minority women living with HIV. AB - BACKGROUND: Directly disclosing a positive HIV serostatus to family members can have psychological and physiological health benefits. Perceptions that one is in a supportive family environment may enhance these benefits. PURPOSE: We examined a mediated moderation model in which we expected interactions between serostatus disclosure to family members and HIV-specific family support to be associated with women's perceived stress, which in turn would explain depressive symptoms and 24-h urinary cortisol in women living with HIV (WLWH). METHOD: Low-income ethnic minority WLWH (n = 82) reported the percentage of family members they had directly disclosed their serostatus to, perceptions of HIV-related support from family members, perceived stress, and depressive symptoms. Cortisol was measured via 24-h urinary collection. RESULTS: Disclosure to spouses and children coupled with high levels of family support was associated with higher levels of depressive symptoms in women. For disclosure to spouses, this relationship was explained by higher perceived stress. Direct disclosure to mothers in tandem with high support was associated with lower cortisol, and this relationship was explained through higher levels of perceived stress. CONCLUSION: The effects of serostatus disclosure on perceived stress and health in WLWH may depend, in part, on women's family environment and to whom they disclose to within that environment. PMID- 19306065 TI - Important roles of the conserved linker-KKS in human neuronal growth inhibitory factor. AB - Metallothinein-3 (MT3), also named neuronal growth inhibitory factor (GIF), is attractive by its distinct neuronal growth inhibitory activity, which is not shared by other MT isoforms. The polypeptide chain of GIF is folded into two individual domains, which are connected by a highly conserved linker, KKS. In order to figure out the significance of the conserved segment, we constructed several mutants of human GIF (hGIF), including the K31/32A mutant, the K31/32E mutant and the KKS-SP mutant by site-directed mutagenesis. pH titration and DTNB reaction exhibited that all the three mutations made the beta-domain lower in stability and looser. More significantly, change of KKS to SP also altered the general backbone conformation and metal-thiolate cluster geometry. Notably, bioassay results showed that the bioactivity of the K31/32A mutant and the K31/32E mutant decreased obviously, while the KKS-SP mutant lost inhibitory activity completely. Based on these results, we proposed that the KKS linker was a crucial factor in modulating the stability and the solvent accessibility of the Cd(3)S(9) cluster in the beta-domain through domain-domain interactions, thus was indispensable to the biological activity of hGIF. PMID- 19306064 TI - Is brachial artery flow-mediated dilation associated with negative affect? AB - BACKGROUND: Trait negative affect has been implicated as a risk marker for cardiovascular disease, but the mechanisms underlying this association are uncertain. PURPOSE: Our aim was to examine associations between trait measures of anger, hostility, depression, and anxiety with endothelial dysfunction via brachial artery flow-mediated dilation (FMD), an early indicator of cardiovascular disease. METHOD: FMD was examined in 332 healthy older adults. Measures included Beck Anxiety and Depression Inventories, Cook-Medley Hostility Scale, and Spielberger State-Trait Anger Expression Inventory (Anger In, Anger Out, and Trait Anger). RESULTS: Mean age was 60.5 +/- 4.8 years; 83% of participants were Caucasian and 49% were female. FMD was greater in women compared to men (6.17% vs. 4.07%, p < 0.001). Women reported significantly greater Anxiety (p < 0.001), and men reported greater Hostility (p = 0.004). In separate multivariable linear regression models controlling for cardiovascular risk factors, plus current hormone therapy for women, smaller FMD was associated with higher Anger In for women (beta = -0.222, p = 0.04) and showed a trend with higher Hostility for men (beta = -0.082, p = 0.09). CONCLUSION: Endothelial dysfunction, as indicated by less vasodilatation of the brachial artery, is positively associated with measures of hostility and anger suppression in healthy older adults. Thus, associations between negative affect and cardiovascular health may be apparent early in the disease process. PMID- 19306066 TI - Biochemical properties of encapsulated high-density 3-D HepG2 aggregates formed in an ultrasound trap for application in hepatotoxicity studies : Biochemical responses of encapsulated 3-D HepG2 aggregates. AB - This paper describes the alginate encapsulation of preformed high-density 3-D HepG2 cell aggregates that guarantees good maintenance of liver-specific biomarker expression. The process involves forming a high-density (> or =7 x 10(4) cells/aggregate) discoid 3-D cell aggregate in an ultrasound trap, which is subsequently recovered and encapsulated in alginate/CaCl(2) hydrogel. Glucose secretion/consumption, lactate release, detoxifying enzyme capacity, cytokeratin 18 expression as well as hypoxia were characterized in encapsulated 3-D HepG2 aggregates over 10 days in culture. Encapsulated 3-D HepG2 aggregates released glucose into the media, although this ability was exhibited only after 1 day in culture and was subsequently lost over the ensuing 9 days. In contrast, lactate was constantly released into the media. Significantly more lactate was secreted after 3 days in culture indicating a more hypoxic environment and hence a higher rate of anaerobic glycolysis. Aggregates consistently expressed cytokeratin-18. Cytochrome P450-1A1 activity reached a maximum on day 1 of culture followed by a progressive reduction to basal levels, while P450-3A4 activity was up-regulated in a time-dependent manner reaching a peak on day 7 in culture. Glutathione-S transferase activity, on the other hand, was at more physiological levels and remained constant over the 10-day culture period. The ultrasound trap allowed the rapid (within 5 min) generation of uniformly shaped and sized aggregates. The results reported here suggest that ultrasound-formed 3-D HepG2 aggregates can serve as alternative in vitro models providing a quick outlook on toxicity, in a tissue-mimetic manner, thus offering the future option of a cost-effective screening platform for pharmaceutical development. PMID- 19306067 TI - Lifestyle factors and serum androgens among 636 middle aged men from seven countries in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - OBJECTIVE: To evaluate the association between lifestyle and dietary factors and serum concentrations of androgens in middle-aged healthy men. METHODS: We conducted a cross-sectional analysis of the association of lifestyle factors with circulating concentrations of androstenedione (A-dione), 3-alpha-androstanediol glucuronide (A-diol-g), testosterone (T), SHBG (sex hormone-binding globulin), and free testosterone (FT) among 636 men in the European Prospective Investigation into Cancer and Nutrition. RESULTS: Compared with the youngest age group (40-49 years), the oldest (70-79 years) had a higher mean concentration of SHBG (by 44%) and lower mean concentrations of A-diol-g (by 29%) FT (19%). Men in the highest BMI group (> or =29.83 kg/m(2)) had a higher mean A-diol-g concentration (by 38%) and lower mean concentration of T (by 20%) SHBG (29%) compared with the lowest (<24.16 kg/m(2)). Current smokers had higher mean concentrations of T (by 13%), SHBG (14%), and A-dione (15%) compared with never smokers. Physical activity and dietary factors were not associated with androgen concentrations, although men in the highest fifth of alcohol intake had higher mean concentrations of A-dione (by 9%), FT (11%) compared with the lowest. CONCLUSION: Our results suggest that age, body weight, smoking, and alcohol intake are associated with circulating androgen concentrations in men. PMID- 19306068 TI - Haematology and leucocyte enzyme cytochemistry of a threatened yellow catfish Horabagrus brachysoma (Gunther 1864). AB - The haematology and leucocyte enzyme cytochemistry of Horabagrus brachysoma, a threatened freshwater catfish endemic to southern India, was studied using standard methods. Intra-specific variation was found for the haematological parameters, but this did not exceed the range of values observed in other catfishes. The relatively high haemoglobin (Hb) concentration may be indicative of an ability to breathe air and high activity. The erythrocytes are fully packed with Hb, revealing the bottom dwelling habit and primitive nature of this catfish. The leucocyte enzyme pattern also showed some variations from those of other fishes. Lymphocytes were positive only for peroxidase (PER) enzyme activity and negative for alkaline phosphatase (LAP), alpha-naphthyl acetate esterase (ANAE) and naphthol ASD chloroacetate esterase (ASDE). Monocytes were weakly positive for ANAE activity and negative for the other three enzymes tested. Neutrophils were negative for LAP, ANAE and ASDE but showed a moderately strong positive reaction for PER. Basophils and eosinophils were found to be devoid of all of these enzymes. Thrombocytes were observed to have weakly positive PER and ASDE, but there was no demonstrable LAP and ANAE activity. A number of characteristics were identified that distinguish this species from other fishes: (1) lymphocytes of H. brachysoma are actively engaged in both phagocytosis and defence mechanisms, while the monocytes participate in cellular defence mechanisms, primarily phagocytosis; (2) thrombocytes function as a protection barrier as well as carrying out their normal function of haemato plug formation during blood clotting. Results from the haematological and leucocyte cytochemical analyses reveal the haematological make-up and effective immune mechanism of this threatened fish and show it to be highly adaptive in nature. The data may be useful in programmes aiming the effective conservation of this species. PMID- 19306069 TI - Automation of cell line development. AB - An automated platform for development of high producing cell lines for biopharmaceutical production has been established in order to increase throughput and reduce development costs. The concept is based on the Cello robotic system (The Automation Partnership) and covers screening for colonies and expansion of static cultures. In this study, the glutamine synthetase expression system (Lonza Biologics) for production of therapeutic monoclonal antibodies in Chinese hamster ovary cells was used for evaluation of the automation approach. It is shown that the automated procedure is capable of producing cell lines of equal quality to the traditionally generated cell lines in terms of colony detection following transfection and distribution of IgG titer in the screening steps. In a generic fed-batch evaluation in stirred tank bioreactors, IgG titers of 4.7 and 5.0 g/L were obtained for best expressing cell lines. We have estimated that the number of completed cell line development projects can be increased up to three times using the automated process without increasing manual workload, compared to the manual process. Correlation between IgG titers obtained in early screens and titers achieved in fed-batch cultures in shake flasks was found to be poor. This further implies the benefits of utilizing a high throughput system capable of screening and expanding a high number of transfectants. Two concentrations, 56 and 75 muM, of selection agent, methionine sulphoximine (MSX), were applied to evaluate the impact on the number of colonies obtained post transfection. When applying selection medium containing 75 muM MSX, fewer low producing transfectants were obtained, compared to cell lines selected with 56 muM MSX, but an equal number of high producing cell lines were found. By using the higher MSX concentration, the number of cell line development projects run in parallel could be increased and thereby increasing the overall capacity of the automated platform process. PMID- 19306070 TI - Erroneous administration of vinblastine. AB - This case describes a series of errors which resulted in an avoidable death of the patient. Upon being presented with the 83-year-old patient and her complaints, the physician in charge attempted to prescribe Vasolastine (a complex preparation used, for example, in treatment of angiopathy, which is administered intramuscularly once a day). Unfortunately he misspelled the name of the medicine as Vinplastyna--a non-existent preparation. When the patient's daughter went to collect the prescription from the pharmacist she was dispensed Vinblastin (vinblastine--a cytostatic medicine used, for example, in treating Hodgkin's disease, non-Hodgkin's lymphoma, chronic lymphatic leukemia and testicular cancer). The visiting community nurses administered a dose of this medicine on seven consecutive days. Upon being given the seventh dose, the patient displayed symptoms of myelophthisis, and was admitted to an Intensive Care Ward, where despite the treatment, she died. PMID- 19306071 TI - Medication dispensing errors in a French military hospital pharmacy. AB - OBJECTIVE: To determine the rate and the primary types of medication dispensing errors detected by pharmacists during implementation of a unit dose drug dispensing system. SETTING: The central pharmacy at the Percy French military hospital (France). METHOD: The check of the unit dose medication cassettes was performed by pharmacists to identify dispensing errors before delivering to the care units. From April 2006 to December 2006, detected errors were corrected and recorded into seven categories: unauthorized drug, wrong dosage-form, improper dose, omission, wrong time, deteriorated drug, and wrong patient errors. MAIN OUTCOME MEASURE: Dispensing error rate, calculated by dividing the total of detected errors by the total of filled and omitted doses; classification of recorded dispensing errors. RESULTS: During the study, 9,719 unit dose medication cassettes were filled by pharmacy technicians. Pharmacists detected 706 errors for a total of 88,609 filled and omitted unit doses. An overall error rate of 0.80% was found. There were approximately 0.07 detected dispensing errors per medication cassette. The most common error types were improper dose errors (n = 265, 37.5%) and omission errors (n = 186, 26.3%). Many causes may probably explain the occurrence of dispensing errors, including communication failures, problems related to drug labeling or packaging, distractions, interruptions, heavy workload, and difficulties in reading handwriting prescriptions. CONCLUSION: The results showed that a wide range of errors occurred during the dispensing process. A check performed after the initial medication selection is also necessary to detect and correct dispensing errors. In order to decrease the occurrence of dispensing errors, some practical measures have been implemented in the central pharmacy. But because some dispensing errors may remain undetected, there is a requirement to develop other strategies that reduce or eliminate these errors. The pharmacy staff is widely involved in this duty. PMID- 19306072 TI - Genes, patients, families, doctors-mutation analysis in clinical practice. AB - Developments in mutation analysis have led to significant benefits for patients with inherited metabolic disorders and their families. This is particularly the case where new methodologies have prevented the need for invasive tissue biopsies or have allowed carrier detection or first trimester prenatal testing to be undertaken. Whereas in the past it may have only been possible to identify specific 'common' mutations, the availability of techniques, such as automated sequencing, and novel technologies including mutation scanning techniques, multiplex ligation dependent probe amplification, and array technologies, have vastly improved the diagnostic efficiency of molecular testing. PMID- 19306074 TI - P2 purinoceptors and the kidney. PMID- 19306073 TI - KCNQ1 loss-of-function mutation impairs gastric acid secretion in mice. AB - The KCNQ1 channel is abundantly expressed in the gastric parietal cells. Although the functional coupling of KCNQ1 with the H(+)/K(+)-ATPase has already been confirmed on the basis of pharmacological kinetics, the effect of a KCNQ1 loss-of function mutation on gastric acidification remains unclear. In this study, parietal cells and gastric glands from both C57BL/6 J mice (normal control) and J343 mice (mice with a KCNQ1 loss-of-function mutation) were isolated to study the effects of KCNQ1 on gastric acidification. We found that the mutation limited intracellular acidification of parietal cells and H(+) secretion of the stomach in response to histamine. Thus, a KCNQ1 loss-of-function mutation may impair gastric acid secretion. PMID- 19306075 TI - ENaC, renal sodium excretion and extracellular ATP. AB - Sodium balance determines the extracellular fluid volume and sets arterial blood pressure (BP). Chronically raised BP (hypertension) represents a major health risk in Western societies. The relationship between BP and renal sodium excretion (the pressure/natriuresis relationship) represents the key element in defining the BP homeostatic set point. The renin-angiotensin-aldosterone system (RAAS) makes major adjustments to the rates of renal sodium secretion, but this system works slowly over a period of hours to days. More rapid adjustments can be made by the sympathetic nervous system, although the kidney can function well without sympathetic nerves. Attention has now focussed on regulatory mechanisms within the kidney, including extracellular nucleotides and the P2 receptor system. Here, we discuss how extracellular ATP can control renal sodium excretion by altering the activity of epithelial sodium channels (ENaC) present in the apical membrane of principal cells. There remains considerable controversy over the molecular targets for released ATP, although the P2Y(2) receptor has received much attention. We review the available data and reflect on our own findings in which ATP-activated P2Y and P2X receptors make adjustments to ENaC activity and therefore sodium excretion. PMID- 19306076 TI - T-cell large granular lymphocytic (T-LGL) leukemia: a single institution experience. AB - BACKGROUND: T-cell large granular lymphocytic (T-LGL) leukemia is a rare lymphoproliferative disease which usually affects elderly people. The clinical course of T-LGL leukemia is generally indolent, with lymphocytosis and splenomegaly in 20-50% patients, hepatomegaly in 5-20% of patients, and less commonly, lymphadenopathy. T-LGL leukemia is associated with immunological abnormalities: rheumatoid factor with or without rheumatoid arthritis (RA), Coombs positive hemolytic anemia, idiopathic thrombocytopenic purpura (ITP), pure red cell aplasia (PRCA), positive anti-nuclear antibodies (ANA), anti-neutrophil cytoplasmic antibodies (ANCA), hypogammaglobulinemia, and polyclonal hypergammaglobulinemia. Aim To compare clinical and laboratory features of T-LGL leukemia patients and their responses to different chemotherapy regimens. METHODS: Six patients (3 males and 3 females) with T-LGL leukemia were analyzed. The diagnosis was based on accepted morphologic criteria, immunophenotype, and polymerase chain reaction (PCR) detection of T-cell receptor (TCR) gene rearrangements. RESULTS: All patients exhibited lymphocytosis, mainly with unusual morphologies, splenomegaly, and elevated serum lactate dehydrogenase (LDH). Three patients were treated with a Fludarabine-Cyclophosphamide (FC) combination as initial therapy while three patients received CHOP. Two patients received more than one treatment regimen. One patient died due to T-LGL leukemia in first year after diagnosis, one patient died 4 years after diagnosis, two patients interrupted their treatment, and two patients are still alive. CONCLUSIONS: Further prospective studies are needed for establishing a gold standard therapy for T-LGL leukemia. PMID- 19306077 TI - 5-Aza-2'-deoxycytidine is a potent inhibitor of DNA methyltransferase 3B and induces apoptosis in human endometrial cancer cell lines with the up-regulation of hMLH1. AB - The aim of our study was to evaluate the effects of 5-aza-2'-deoxycytidine (5 azadC) on cell growth inhibition, cell cycle arrest, apoptosis as well as the expression levels of hMLH1 and DNMT3B in human endometrial cancer cell lines. Ishikawa, HHUA, and KLE cell lines were used. After treatment with 5-azadC, cells were measured by MTT to detect the growth inhibition. Flow cytometry analysis was used to evaluate the cell cycle distribution and apoptosis effect. The expression of hMLH1 and DNMT3B was performed by real-time PCR and Western blotting analysis. The methylation status of the hMLH1 gene was monitored by methylation-specific PCR. We confirmed that 5-azadC treatment resulted in growth inhibition, G(2) arrest, and cell apoptosis in human endometrial cancer cell lines. Furthermore, the data obtained by real-time PCR and Western blotting analysis demonstrated that the expression of hMLH1 was up-regulated by 5-azadC treatment in Ishikawa cells, accompanied by down-regulation of DNMT3B expression, when 5-azadC led to cell inhibition, G(2)/M arrest, and apoptosis. Our results suggested that 5-azadC is a potent inhibitor of DNA methyltransferase 3B and induces apoptosis in Ishikawa cells with the up-regulation of hMLH1. PMID- 19306079 TI - The ultimate "analytical challenge": what is analytical chemistry? PMID- 19306078 TI - High-level expression, purification, and in vitro refolding of soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a new member of the TNF superfamily. In this paper, we report the expression, purification, and preparation of a recombinant form of the extracelluar domain of the TRAIL (sTRAIL) without posttranslational modifications, which may selectively induce apoptosis of tumor cells in vitro. To obtain recombinant nonfusion sTRAIL protein, the encoding region for sTRAIL was cloned between KpnI and BamHI in pET32a. The Trx (thioredoxin)/sTRAIL fusion proteins were expressed in the form of inclusion bodies in Escherichia coli host strain BL21 (DE3). The expression level was more than 35% of total cell lysate. Inclusion bodies were disrupted, washed, and isolated at pH 9.0, and were completely dissolved in a buffer containing 2 M urea at pH 9.0. After nickel ion metal affinity chromatography, gel filtration chromatography, and renaturation, the refolded fusion proteins with a purity of >98% were obtained. Trx/sTRAIL L proteins were digested by enterokinase to both Trx and sTRAIL fragments, which then were separated by cation exchange chromatography. Cell proliferation experiments proved that the rsTRAIL (98% purity) retains its cancer-selective apoptosis-inducing properties. This result suggested that the recombinant sTRAIL may have cancer therapeutic applications. PMID- 19306080 TI - A fiber-optic evanescent wave sensor for dissolved oxygen detection based on novel hybrid fluorinated xerogels immobilized with [Ru(bpy)3]2+. AB - We have prepared a novel fiber-optic evanescent wave sensor (FEWS) for dissolved oxygen (DO) detection. The sensor fabrication was based on coating a decladded portion of an optical fiber with a microporous coating, which was prepared from 3,3,3-trifluoropropyltrimethoxysilane and n-propyltrimethoxysilane. The fluorophores were immobilized in the porous coating and excited by the evanescent wave field produced on the core surface of the optical fiber. The sensitivity of the sensor was quantified by the ratio of the fluorescence intensities in pure deoxygenated (I (0)) and in pure oxygenated environments (I). Results show that the quenching response of DO is increased with the enhancement of the coating surface hydrophobicity using the presented hybrid fluorinated ORMOSILs. The calibration curve of I (0)/I to [O(2)] is linear from 0 to 40 ppm and the detection limit is 0.05 ppm (3sigma) with a short response time of 15 s for DO detection. PMID- 19306081 TI - Assessment of the genotoxicity of endosulfan in earthworm and white clover plants using the comet assay. AB - Endosulfan, as one of the most widely used organochlorine pesticides in the world, has increased the public concern about genotoxicity in soil ecosystems. The comet assay has been widely used in the fields of genetic toxicology and environmental biomonitoring. In the present study we conducted comet assay of endosulfan in earthworm (Eisenia foetida) and white clover (Trifolium repens L.), which are sensitive organisms suitable for acting as a bioindicator for agricultural ecosystems. Earthworms were exposed to endosulfan concentrations of 0.1, 1.0, and 10.0 mg/kg in the soil. White clover roots were immersed in hydroponic pots containing nutrient solutions of different endosulfan concentrations: 0.1, 1.0, and 10.0 mg/L. Tissues from each treatment were collected on the 7th, 14th, 21st, and 28th days of treatment process. Significant effects (p < 0.01) of both concentrations and times of exposure were observed. And endosulfan induced DNA damage in earthworm and white clover nuclei. The comet assay can be used as a reliable tool for early detection of endosulfan. PMID- 19306083 TI - Entrapment in anti myelin-associated glycoprotein neuropathy. AB - Anti-myelin associated glycoprotein (MAG) neuropathy is a chronic disorder in which IgM antibodies react with Schwann cell glycoproteins, including MAG and peripheral myelin protein 22 (PMP22). Nerve conduction studies show features of axon loss and predominantly distal slowing consistent with demyelination. Because a genetic loss of PMP22 function yields hereditary neuropathy with liability to pressure palsies (HNPP), loss of PMP22 function due to anti- MAG antibodies may result in increased sensitivity to entrapment. We investigated this by performing standardized electrophysiological studies in 16 patients with anti-MAG neuropathy and 16 disease controls with genetically confirmed HNPP. Disproportionate slowing relative to adjacent segments occurred in similar proportions of patients with anti-MAG neuropathy and HNPP, and was of the same magnitude in each group. Affected were the elbow, carpal tunnel and the wrist-hand segments of the median and ulnar nerves. However, in anti-MAG neuropathy as compared to HNPP, absolute values of distal motor latencies and conduction velocities outside entrapment sites were slower and amplitudes were lower. In conclusion, increased sensitivity for entrapment may occur in anti-MAG neuropathy and contribute to part of the nerve damage. PMID- 19306085 TI - Improvement of arachidonic acid and eicosapentaenoic acid production by increasing the copy number of the genes encoding fatty acid desaturase and elongase into Pichia pastoris. AB - Genes encoding Delta6 desaturase, Delta6 fatty acid elongase, and Delta5 desaturase from the alga, Phaeodactylum tricornutum, were co-expressed in Pichia pastoris to produce arachidonic acid (ARA; 20:4 Delta(5, 8, 11, 14)) and eicosapentaenoic acid (EPA; 20:5 Delta(5, 8, 11, 14, 17)). A panel of Pichia clones carrying progressively increasing copies of the heterologous gene expression cassette was created using an in vitro multimerization approach. ARA and EPA accumulated up to 0.3 and 0.1% of total fatty acids, respectively, in the recombinant P. pastoris carrying with double copies of these three heterologous genes, as compared to 0.1 and 0.05%, respectively, in the recombinant P. pastoris with single copy. PMID- 19306086 TI - Zinc transport by respiratory epithelial cells and interaction with iron homeostasis. AB - Despite recurrent exposure to zinc through inhalation of ambient air pollution particles, relatively little information is known about the homeostasis of this metal in respiratory epithelial cells. We describe zinc uptake and release by respiratory epithelial cells and test the postulate that Zn(2+) transport interacts with iron homeostasis in these same cells. Zn(2+) uptake after 4 and 8 h of exposure to zinc sulfate was concentration- and time-dependent. A majority of Zn(2+) release occurred in the 4 h immediately following cell exposure to ZnSO(4). Regarding metal importers, mRNA for Zip1 and Zip2 showed no change after respiratory epithelial cell exposure to zinc while mRNA for divalent metal transporter (DMT)1 increased. Western blot assay for DMT1 protein supported an elevated expression of this transport protein following zinc exposure. RT-PCR confirmed mRNA for the metal exporters ZnT1 and ZnT4 with the former increasing after ZnSO(4). Cell concentrations of ferritin increased with zinc exposure while oxidative stress, measured as lipid peroxides, was decreased supporting an anti oxidant function for Zn(2+). Increased DMT1 expression, following pre-incubations of respiratory epithelial cells with TNF-alpha, IFN-gamma, and endotoxin, was associated with significantly decreased intracellular zinc transport. Finally, incubations of respiratory epithelial cells with both zinc sulfate and ferric ammonium citrate resulted in elevated intracellular concentrations of both metals. We conclude that exposure to zinc increases iron uptake by respiratory epithelial cells. Elevations in cell iron can possibly affect an increased expression of DMT1 and ferritin which function to diminish oxidative stress. Comparable to other metal exposures, changes in iron homeostasis may contribute to the biological effects of zinc in specific cells and tissues. PMID- 19306087 TI - Biomarker science: direction for the scientific community. PMID- 19306088 TI - Evaluation of malignancy-risk gene signature in breast cancer patients. AB - We recently developed a malignancy-risk gene signature that was shown to identify histologically-normal tissues with a cancer-like profile. Because the signature was rich with proliferative genes, we postulated it might also be prognostic for existing breast cancers. We evaluated the malignancy risk gene signature to see its clinical association with cancer relapse/progression, and cancer prognosis using six independent external datasets. Six independent external breast cancer datasets were collected and analyzed using the malignancy risk gene signature designed to assess normal breast tissues. Evaluation of the signature in external datasets suggested a strong clinical association with cancer relapse/progression, and prognosis with minimal overlap of signature gene sets. These results suggest a prognostic role for the malignancy risk gene signature in the assessment of existing cancer. Proliferative biology dominates not only the earliest stages of tumor development but also later stages of tumor progression and metastasis. PMID- 19306089 TI - AAV-mediated local delivery of interferon-beta for the treatment of retinoblastoma in preclinical models. AB - Interferon-beta (IFN-beta) has been found to have anti-tumor properties against a variety of malignancies through different mechanisms. However, clinical trials involving systemic administration of IFN-beta have been hampered by secondary toxicity and the short half-life of IFN-beta in the circulation. In order to circumvent these limitations, we have developed an adeno-associated viral (AAV) vector gene-therapy approach to deliver IFN-beta to tumors. In this study, we tested the efficacy of AAV-mediated local delivery of IFN-beta for the treatment of retinoblastoma in preclinical models. Retinoblastoma is an ideal candidate for gene-therapy-based anti-cancer treatment because target cell transduction and, therefore, IFN-beta delivery can be contained within the ocular environment, thereby minimizing systemic toxicity. We report here that retinoblastoma cell lines exhibit pleiotropic responses to IFN-beta consistent with previous studies on a variety of tumor cell lines. Intravitreal injection of AAV-IFN-beta resulted in efficient retinal infection and sustained IFN-beta production in the eye with minimal systemic exposure. Vector spread outside of the eye was not detected. Using our orthotopic xenograft model of retinoblastoma, we found that intravitreal injection of AAV-IFN-beta had a potent anti-tumor effect in vivo. These data suggest that AAV-mediated delivery of IFN-beta may provide a complementary approach to systemic chemotherapy which is the standard of care for retinoblastoma around the world. PMID- 19306090 TI - Influence of visually guided tracking arm movements on single cell activity in area MT. AB - The behavioral relevance of neuronal activity in primate area MT for motion perception and control of visually guided eye movements is well documented. The projections of area MT comprise connections to subcortical structures and to the parietal network, both of which play a role in visuospatial transformation for guiding eyes and hands. Here, we have investigated, whether area MT is involved in the network needed to control visually guided arm movements. Our results show that half of the neurons tested significantly modulated their activity during visually guided arm movements. We conclude that the main reason for the neuronal modulation is not the arm movement per se, but the use of information from MT for visual feedback in the tracking movement. Moreover, control experiments show that attentional effects cannot solely cause the neuronal modulation. Thus, our study provides strong evidence that area MT is involved in processing visual information for visually guided manual tracking movements. PMID- 19306091 TI - Specificity of auditory-guided visual perceptual learning suggests crossmodal plasticity in early visual cortex. AB - Sounds modulate visual perception. Blind humans show altered brain activity in early visual cortex. However, it is still unclear whether crossmodal activity in visual cortex results from unspecific top-down feedback, a lack of visual input, or genuinely reflects crossmodal interactions at early sensory levels. We examined how sounds affect visual perceptual learning in sighted adults. Visual motion discrimination was tested prior to and following eight sessions in which observers were exposed to irrelevant moving dots while detecting sounds. After training, visual discrimination improved more strongly for motion directions that were paired with a relevant sound during training than for other directions. Crossmodal learning was limited to visual field locations that overlapped with the sound source and was little affected by attention. The specificity and automatic nature of these learning effects suggest that sounds automatically guide visual plasticity at a relatively early level of processing. PMID- 19306092 TI - Mode of action of cannabinoids on nociceptive nerve endings. AB - In recent years, cannabinoids have emerged as attractive alternatives or supplements to therapy for chronic pain states. However, in humans the activation of cannabinoid receptors in neurons of the central nervous system is associated with psychotropic side effects, temporary memory impairment and dependence, which arise via the effects of cannabinoids on forebrain circuits. For clinical exploitation of the analgesic properties of cannabinoids, a major challenge is to devise strategies that reduce or abolish their adverse effects on cognitive, affective and motor functions without attenuating their analgesic effects. The cannabinoid receptor family currently includes two cloned metabotropic receptors: CB1, CB2 and possibly GPR55 which are distributed widely across many key loci in pain-modulating pathways, including the peripheral terminals of primary afferents. Modulation of transducer ion channels expressed at nociceptive terminals occurs upon activation of metabotropic cannabinoid receptors, but direct cannabinoid action on ion channels involved in sensory transduction or regulation of neuron excitability likely contributes to the peripheral cannabinoid effects. PMID- 19306093 TI - Thymidylate synthetase allelic imbalance in clear cell renal carcinoma. AB - PURPOSE: To investigate the allelic status of the thymidylate synthetase (TYMS) gene, located at chromosome band 18p11.32, in renal cell carcinoma (RCC). TYMS is a key target of the 5-fluorouracil (5-FU)-based class of drugs, frequently considered in combination therapies in advanced RCC. TYMS variants, such as the TYMS polymorphic 5'-untranslated region variable number tandem repeat sequence (VNTR), are under investigation to guide 5-FU treatment. Yet, no information is available with regard to changes in TYMS allele frequencies in RCC malignances. METHODS: Blood and matched tumor samples were collected from 41 histological proven clear cell RCC affected patients (30 males, 11 females.). TYMS VNTR genotype was first determined in blood to identify heterozygotes employing PCR techniques. To evaluate for allelic imbalance, fragment analysis was performed both in blood and matched tumor DNA of the heterozygote patients. Microsatellite analysis, employing the markers D18S59 and D18S476 mapping, respectively, at the TYMS locus (18p11.32) and 1.5 Mb downstream of the TYMS gene sequence (18p11.31), was performed to confirm TYMS allelic imbalance in tumors. RESULTS: Germ-line TYMS VNTR distribution was: 2R/2R (19.5%), TYMS 2R/3R (36.6%) and TYMS 3R/3R (43.9%). Allelic imbalance for the TYMS tandem repeat region was detected in 26.6% of the heterozygote patients. Microsatellite analysis confirmed the allelic imbalance detected by TYMS VNTR analysis and revealed that the overall frequence of allelic imbalance of chromosome band 18p11.32 was 35%, while the overall allelic imbalance of chromosome band 18p11.31 was 28%. CONCLUSIONS: By focusing on the TYMS polymorphic variants in renal cancer, we here provide evidence, to our knowledge, for the first time showing loss of 18p11.32 and 18p11.31 in renal cell carcinomas. As allelic imbalances involving TYMS locus may be an important variable affecting 5-FU responsiveness, this study may contribute to explain different responses of advanced RCC in combined chemotherapeutic regimens incorporating fluoropyridines. PMID- 19306094 TI - Schedule-dependent synergism and antagonism between pemetrexed and docetaxel in human lung cancer cell lines in vitro. AB - BACKGROUND: Pemetrexed and docetaxel show clinical activities against a variety of solid tumors including lung cancers. To identify the optimal schedule for combination, cytotoxic interactions between pemetrexed and docetaxel were studied at various schedules using three human lung cancer cell lines A-549, Lu-99, and SBC-5 in vitro. METHODS: Cells were incubated with pemetrexed and docetaxel simultaneously for 24 or 120 h. Cells were also incubated with pemetrexed for 24 h, followed by a 24 h exposure to docetaxel, and vice versa. Growth inhibition was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell cycle analysis. Cytotoxic interactions were evaluated by the isobologram method. RESULTS: Simultaneous exposure to pemetrexed and docetaxel for 24 and 120 h produced antagonistic effects in all three cell lines. Pemetrexed (24 h) followed by docetaxel (24 h) produced additive effects in A-549 cells and synergistic effects in Lu-99 and SBC-5 cells. Docetaxel followed by pemetrexed produced additive effects in A-549 and Lu-99 cells and antagonistic effects in SBC-5 cells. The results of cell cycle analysis were fully consistent with those of isobologram analysis, and provide the molecular basis of the sequence-dependent difference in cytotoxic interactions between the two agents. CONCLUSIONS: Sequential administration of pemetrexed followed by docetaxel may provide the greatest anti-tumor effects for this combination in the treatment of lung cancer. PMID- 19306095 TI - Homozygous mutations in the 15-hydroxyprostaglandin dehydrogenase gene in patients with primary hypertrophic osteoarthropathy. AB - Mutations in HPGD have recently been reported to cause primary hypertrophic osteoarthropathy (PHO), a rare genetic disease characterized by digital clubbing, pachydermia, and periostosis. We screened HPGD mutations in six patients from three unrelated Turkish families with PHO, in which we showed one previously reported, p.A140P, and one novel, p.M1L, homozygous mutations. Both mutations co segregated with the phenotype in all three families and were absent in 100 Turkish controls. These results confirm the presence of biallelic HPGD mutations in patients with PHO in an independent series from a different population. PMID- 19306096 TI - The ubiquitin-proteasome pathway and viral infections in articular cartilage of patients with osteoarthritis. AB - Many viruses can evolve different strategies to exploit the ubiquitin-proteasome pathway (UPP) for their own benefit. Some data have recently established connections between UPP and osteoarthritis (OA). The aim of this study was to determine the possible involvement of viral infections linked with the UPP in the physiopathology of OA. Samples of human cartilage were obtained from 12 patients with clinical and radiological features of OA and from 12 normal controls. DNA was extracted from cultured chondrocytes from these patients, and quantitative real-time PCR was performed to analyse the DNA/RNA prevalence and viral loads of HSV, EBV, HCMV, enterovirus, and HTLV-1. The prevalence of total viral DNA/RNA among patients with OA was 16.7% (mean viral load of 7.86 copies/mug DNA), EBV being responsible for the two positive samples, while the prevalence in controls was 0%. We did not detect any positive samples for HSV, CMV, enterovirus, and HTLV-1 among patients with OA and controls. This first approach to the study of the prevalence of viruses linked to the UPP in articular cartilage of end-stage OA patients provides evidences supporting the risk of EBV transmission or reactivation in a subset of patients with disorders requiring tissue regeneration. PMID- 19306097 TI - Neuropsychiatric manifestations and antiphospholipid antibodies in pediatric onset lupus: 14 years of experience from a tertiary center of North India. AB - The objective is to study the neuropsychiatric (NP) manifestations in pediatric onset systemic lupus erythematosus (SLE) at a tertiary care hospital of northwestern India applying American College of Rheumatology (ACR) case definitions in the context of occurrence of antiphospholipid antibodies (APLA). Data of 53 children with SLE were analyzed for NP syndromes. Tests for detection of APLA were performed as per international standards for quality control. Twenty seven of the 53 (50.94%) children with lupus had at least one NP manifestation. The male to female ratio of our cohort of pediatric lupus was 1:2.8. However, there was significant male preponderance in patients with NPSLE as compared to patients without NPSLE (1:1.25 vs. 1:12; P < 0.0001). Majority of children with NPSLE (15/27, 55.5%) already had NP manifestations at the time of diagnosis and most of them (81.5%) had experienced more than one NP symptom. Headache was the commonest NP manifestation and was seen in 39.6% children with SLE followed by seizure disorder (35.8%) and cognitive dysfunction (16.9%). Tests for APLA were carried out in 37 of 53 (69.8%) patients with SLE and in 24 of 27 (88.8%) patients with NPSLE. While anticardiolipin antibodies were seen more frequently in children with NPSLE as compared to those without NPSLE (57.8 vs. 23%), lupus anticoagulant was more frequent in children without NPSLE (53.8 vs. 34.7%). However, these differences were statistically not significant. Eleven of the 27 children with NPSLE succumbed to their illness, primarily due to uncontrolled disease activity. Mean duration of follow-up of patients with NPSLE who are alive was 65.4 +/- 36.9 months. NP manifestations are common in pediatric onset lupus and contribute to significant morbidity. As compared to previously published literature, a significantly greater proportion of boys were affected. APLA were frequently detected in children with NPSLE. There is paucity of literature pertaining to NP manifestations of pediatric lupus in the context of APLA, especially with regard to antibodies to beta-2 glycoprotein I. To the best of our knowledge, this is the first detailed study on NP manifestations in childhood lupus from a developing country applying ACR case definitions. PMID- 19306098 TI - Variations in the neuropathology of familial Alzheimer's disease. AB - Mutations in the amyloid precursor protein (APP), presenilin 1 (PSEN1) and presenilin 2 (PSEN2) genes cause autosomal dominant familial Alzheimer's disease (AD). PSEN1 and PSEN2 are essential components of the gamma-secretase complex, which cleaves APP to affect Abeta processing. Disruptions in Abeta processing have been hypothesised to be the major cause of AD (the amyloid cascade hypothesis). These genetic cases exhibit all the classic hallmark pathologies of AD including neuritic plaques, neurofibrillary tangles (NFT), tissue atrophy, neuronal loss and inflammation, often in significantly enhanced quantities. In particular, these cases have average greater hippocampal atrophy and NFT, more significant cortical Abeta42 plaque deposition and more substantial inflammation. Enhanced cerebral Abeta40 angiopathy is a feature of many cases, but particularly those with APP mutations where it can be the dominant pathology. Additional frontotemporal neuronal loss in association with increased tau pathology appears unique to PSEN mutations, with mutations in exons 8 and 9 having enlarged cotton wool plaques throughout their cortex. The mechanisms driving these pathological differences in AD are discussed. PMID- 19306100 TI - Fracture of maternal sternum during spontaneous delivery. AB - We report of a maternal sternal fracture during a spontaneous delivery in a 31 year-old primipara without a suitable trauma. The putative mechanism of fracture was strong hyperflexion of the thoracic spine and additional cervical flexion with pushing the chin to the thorax due to active management of labour. The history of the healthy woman was free of related risk factors. A possible promoting factor might be pregnancy-induced bone loss. Although there were clear symptoms, the diagnosis of the fracture was delayed by a week because nobody took account of such a possibility. PMID- 19306099 TI - Overexpression of manganese superoxide dismutase in human dermal fibroblasts enhances the contraction of free floating collagen lattice: implications for ageing and hyperplastic scar formation. AB - Cell-matrix interactions are of significant importance for tissue homeostasis of the skin and, if disturbed, may lead to ageing and hyperplastic scar formation. We have studied fibroblasts stably overexpressing manganese superoxide dismutase (MnSOD) with a defined capacity for the removal of superoxide anions and concomitant accumulation of hydrogen peroxide to evaluate the role of enhanced MnSOD activity on the dynamics of cell-matrix interactions in the three dimensional collagen lattice contraction assay. MnSOD overexpressing fibroblast populated collagen lattices revealed a significantly enhanced contraction compared to collagen lattices populated with vector control cells. The enhanced collagen lattice contraction was in part due to an increase in active TGF-beta1 and the accumulation of H2O2 in MnSOD overexpressing fibroblasts populated collagen lattices. Inhibition of TGF-beta1 signalling by the ALK4,5,7 kinases' inhibitor SB431542 at least partly inhibited the enhanced collagen lattice contraction of MnSOD overexpressing fibroblasts populated lattices. In addition, supplementation of vector control fibroblast populated collagen lattices with recombinant TGF-beta1 concentration dependently enhanced the collagen lattice contraction. In the presence of the antioxidant Ebselen, a mimic of H2O2 and other hydroperoxides/peroxynitrite-detoxifying glutathione peroxidase, collagen lattice contraction and the activation of TGF-beta1 were significantly reduced in collagen lattices populated with MnSOD overexpressing fibroblasts. Collectively, these data suggest that H2O2 or other hydroperoxides or peroxynitrite or a combination thereof may function as important second messengers in collagen lattice contraction and act at least in part via TGF-beta1 activation. PMID- 19306101 TI - Coexistent tubercular cervicitis with tuberculosis of the little finger: an unusual presentation. PMID- 19306102 TI - A MYB transcription factor regulates anthocyanin biosynthesis in mangosteen (Garcinia mangostana L.) fruit during ripening. AB - Mangosteen (Garcinia mangostana L.) fruit undergo rapid red colour development, both on the tree and after harvest, resulting in high anthocyanin production in the pericarp. Here, we report the isolation of three full-length mangosteen MYB transcription factors (GmMYB1, GmMYB7 and GmMYB10) and all the anthocyanin biosynthetic pathway genes (GmPal to GmUFGT). Phylogenetic analysis at the protein level of the R2R3-MYB transcription factor family showed GmMYB10 had a high degree of similarity with production of anthocyanin pigment1 in Arabidopsis and as well as sequences from other plant species related to the elevation of anthocyanin pigmentation. In transient transactivation assays, GmMYB10, co expressed with AtbHLH2, strongly activated the GmDFR and AtDFR promoters. Transcripts of GmMYB10 and GmUFGT were highly abundant with onset of pigmentation and subsequently during red colouration. Our results suggest that GmMYB10 plays an important role in regulating anthocyanin biosynthesis both on the tree and after harvest, while GmUFGT may be a key biosynthetic gene in mangosteen pigmentation. The expression patterns of GmMYB10 and GmUFGT correlated with ethylene production that increased linearly until stage 5 (dark purple) and decreased thereafter. 1-Methycyclopropene (1-MCP) clearly delayed red colouration with resulting down-regulation of GmMYB10. These results suggest that the effect of ethylene on anthocyanin biosynthesis may be via the regulation of GmMYB10 expression. PMID- 19306103 TI - CYP710A genes encoding sterol C22-desaturase in Physcomitrella patens as molecular evidence for the evolutionary conservation of a sterol biosynthetic pathway in plants. AB - We have characterized cytochromes P450, CYP710A13, and CYP710A14, as the sterol C22-desaturase in the moss Physcomitrella patens. GC-MS analyses demonstrated that P. patens accumulated stigmasterol as the major sterol (56-60% of total sterol) and sitosterol to a lesser extent (8-12%); this sterol profile contrasts with those in higher plants accumulating stigmasterol as a minor component. Recombinant CYP710A13 and CYP710A14 proteins prepared using a baculovirus/insect cell system exhibited the C22-desaturase activity with beta-sitosterol to produce stigmasterol, while campesterol and 24-epi-campesterol were not accepted as the substrates. The K(m) values for beta-sitosterol of CYP710A13 (1.0 +/- 0.043 microM) and CYP710A14 (2.1 +/- 0.17 microM) were at comparable levels of those reported with higher plant CYP710A proteins. In Arabidopsis T87 cells over expressing CYP710A14, stigmasterol contents reached a level 20- to 72-fold higher than those in the basal level of T87 cells, confirming the C22-desaturase activity of this P450 enzyme. The occurrence of the end-products together with the enzymes involved in the last step of the pathway substantiated the presence of an entire sterol biosynthetic pathway in P. patens, providing evidence for the conservation of the sterol biosynthetic pathway through the evolutionary process of land plants. PMID- 19306104 TI - The short-rooted vitamin B6-deficient mutant pdx1 has impaired local auxin biosynthesis. AB - The phytohormone auxin regulates many aspects of plant growth and development. Auxin often acts distantly from the site of its biosynthesis and this long distance-transported auxin is well known to play a critical role in eliciting physiological responses including regulating root development. Auxin can be produced in roots, yet the function of locally synthesized auxin in root growth is unclear. The major auxin in plants, indole 3-acetic acid (IAA), is mainly synthesized through tryptophan (Trp)-dependent pathways that require pyridoxal phosphate (an active form of vitamin B(6))-dependent enzymes. We previously reported that the Arabidopsis vitamin B(6) biosynthesis mutant pdx1 has stunted root growth although the underlying cause is unknown. Here we showed that the pdx1 root is deficient in auxin biosynthesis. By reciprocal grafting of pdx1 and the wild type, we demonstrated that the stunted root growth in pdx1 is caused by a locally generated signal(s) in roots. To test whether auxin might be one such signal, the auxin responsive DR5::GUS reporter was introduced into the mutant. The DR5::GUS activity in pdx1 root tips was greatly reduced compared with that in the wild type although the auxin response was unaltered. pdx1 also suppresses the root hair growth defects in the auxin overproduction mutant yucca. These data indicate that pdx1 is impaired in Trp-dependent auxin biosynthesis, which may contribute to the short-root phenotype of pdx1. We suggest that locally synthesized auxin may play a critical role in postembryonic root growth. PMID- 19306105 TI - A model for the spatiotemporal organization of DNA replication in Saccharomyces cerevisiae. AB - DNA replication in eukaryotes is considered to proceed according to a precise program in which each chromosomal region is duplicated in a defined temporal order. However, recent studies reveal an intrinsic temporal disorder in the replication of yeast chromosome VI. Here we provide a model of the chromosomal duplication to study the temporal sequence of origin activation in budding yeast. The model comprises four parameters that influence the DNA replication system: the lengths of the chromosomes, the explicit chromosomal positions for all replication origins as well as their distinct initiation times and the replication fork migration rate. The designed model is able to reproduce the available experimental data in form of replication profiles. The dynamics of DNA replication was monitored during simulations of wild type and randomly perturbed replication conditions. Severe loss of origin function showed only little influence on the replication dynamics, so systematic deletions of origins (or loss of efficiency) were simulated to provide predictions to be tested experimentally. The simulations provide new insights into the complex system of DNA replication, showing that the system is robust to perturbation, and giving hints about the influence of a possible disordered firing. PMID- 19306106 TI - Population admixture modulates risk for alcohol dependence. AB - The admixture of different ancestral populations in America may have important implications for the risk for psychiatric disorders, as it appears to have for other medical disorders. The present study investigated the role of population admixture in risk for several psychiatric disorders in European-Americans (EAs) and African-Americans (AAs). This is a multisite study with 3,792 subjects recruited from across the United States, including 3,119 EAs and 673 AAs. These subjects included healthy controls and those with substance dependence (SD) [including alcohol dependence (AD), cocaine dependence, and opioid dependence], social phobia, affective disorders, and schizophrenia. In addition, DNA was included from 78 West Africans. The degree of admixture for each subject was estimated by analysis of a set of ancestry-informative genetic markers using the program STRUCTURE, and was compared between cases and controls. As noted previously, the degree of admixture in AAs was higher than EAs. In EAs, the degree of admixture (with African ancestry) was significantly lower in patients with SD (mainly AD) than controls (P = 0.009 for SD; P = 0.008 for AD). This finding suggests that population admixture may modulate risk for alcohol dependence. Population admixture might protect against alcohol dependence by increasing average heterozygosity and reducing the risk of deleterious recessive alleles. We cannot exclude the possibility that the results might have been influenced by selection bias due to the multisite nature of the study. PMID- 19306107 TI - Quantifying the dose-response of walking in reducing coronary heart disease risk: meta-analysis. AB - The evidence for the efficacy of walking in reducing the risk of and preventing coronary heart disease (CHD) is not completely understood. This meta-analysis aimed to quantify the dose-response relationship between walking and CHD risk reduction for both men and women in the general population. Studies on walking and CHD primary prevention between 1954 and 2007 were identified through Medline, SportDiscus and the Cochrane Database of Systematic Reviews. Random-effect meta regression models were used to pool the relative risks from individual studies. A total of 11 prospective cohort studies and one randomized control trial study met the inclusion criteria, with 295,177 participants free of CHD at baseline and 7,094 cases at follow-up. The meta-analysis indicated that an increment of approximately 30 min of normal walking a day for 5 days a week was associated with 19% CHD risk reduction (95% CI = 14-23%; P-heterogeneity = 0.56; I (2) = 0%). We found no evidence of heterogeneity between subgroups of studies defined by gender (P = 0.67); age of the study population (P = 0.52); or follow-up duration (P = 0.77). The meta-analysis showed that the risk for developing CHD decreases as walking dose increases. Walking should be prescribed as an evidence based effective exercise modality for CHD prevention in the general population. PMID- 19306108 TI - Arylsulfatase B regulates colonic epithelial cell migration by effects on MMP9 expression and RhoA activation. AB - Arylsulfatase B (ASB; N-acetylgalactosamine-4-sulfatase; 4-sulfatase; ARSB) is the enzyme that removes 4-sulfate groups from N-acetylgalactosamine 4-sulfate, which combines with glucuronate to form the disaccharide unit of chondroitin-4 sulfate (C4S). In this study, we report how variation in expression of ASB affected the migration of human colonic epithelial cells. In the T84 cell line, derived from lung metastasis of malignant colonic epithelial cells, the activity of ASB, as well as steroid sulfatase, arylsulfatase A, and galactose-6-sulfatase, were significantly less than in normal, primary colonic epithelial cells and in the NCM460 cell line which was derived from normal colonocytes. In the T84 cells, matrix metalloproteinase 9 (MMP9), activated RhoA, and cell migration, as well as C4S content, were significantly more than in the NCM460 cells. Silencing and overexpression of ASB had inverse effects on MMP9, activated RhoA, and cell migration, as well as the C4S content, in the NCM460 and T84 cells. When ASB expression was silenced by siRNA in the NCM460 cells, MMP9 secretion increased to over 3 times the basal level, activated RhoA increased * 85%, and cell migration increased * 52%. Following overexpression of ASB, MMP9 declined 51%, activated RhoA declined * 51%, and cell migration decreased * 37%. These findings demonstrate marked effects of ASB expression on the migratory activity of colonic epithelial cells, activated RhoA, and MMP9, and suggest a potential vital role of ASB, due to its impact on chondroitin sulfation, on determination of the invasive phenotype of colonic epithelial cells. PMID- 19306110 TI - Characterization of the beneficial properties of lactobacilli isolated from bullfrog (Rana catesbeiana) hatchery. AB - The present work addresses the isolation and partial identification of the microbial population of a R. catesbeiana hatchery in spring and summer as well as some beneficial properties of Lactobacillus strains isolated in different seasons and hatchery areas. The bacterial population was grouped into the following taxa: Lactobacillus spp., Pediococcus spp., Enterococcus faecalis and Ent. faecium, and Enterobacteriaceae (Enterobacter spp., Escherichia coli) while Pseudomonas aeruginosa and Staphylococcus epidermidis were isolated from frogs displaying red leg syndrome. The Lactobacillus plantarum and L. curvatus strains isolated showed to inhibit the growth of red-leg syndrome associated pathogens and food-borne bacteria by organic acids. While L. plantarum CRL 1606 also inhibited red-leg syndrome related pathogens by hydrogen peroxide, meat spoilage bacteria were only inhibited by acidity. However, by using a MRS medium added with tetramethyl benzidine and peroxidase, a high percentage of H(2)O(2)-producing lactobacilli were detected. The surface properties of Lactobacillus strains showed that a few strains were able to agglutinate ABO human erythrocytes, while the highest number of strains had a low to medium degree of hydrophobicity. This paper constitute the first study related to the beneficial properties of Lactobacillus isolated from a bullfrog hatchery, as well as the selection criteria applied to a group of strains, which could help to control or prevent bacterial infectious diseases in raniculture. PMID- 19306109 TI - Regulator of calcineurin 1 modulates cancer cell migration in vitro. AB - Metastasis suppressors and other regulators of cell motility play an important role in tumor invasion and metastases. We previously identified that activation of the G protein coupled receptor 54 (GPR54) by the metastasis suppressor metastin inhibits cell migration in association with overexpression of Regulator of calcineurin 1 (RCAN1), an endogenous regulator of calcineurin. Calcineurin inhibitors also blocked cell migration in vitro and RCAN1 protein levels were reduced in nodal metastases in thyroid cancer. The purpose of the current study was to determine directly if RCAN1 functions as a motility suppressor in vitro. Several cancer cell lines derived from different cancer types with different motility rates were evaluated for RCAN1 expression levels. Using these systems we determined that reduction of endogenous RCAN1 using siRNA resulted in an increase in cancer cell motility while expression of exogenous RCAN1 reduced cell motility. In one cell line with a high migratory rate, the stability of exogenously expressed RCAN1 protein was reduced and was rescued by treatment with a proteasome inhibitor. Finally, overexpression of RCAN1 was associated with an increase in cell adhesion to collagen IV and reduced calcineurin activity. In summary, we have demonstrated that the expression of exogenous RCAN1 reduces migration and alters adhesion; and that the loss of endogenous RCAN1 leads to an increase in migration in the examined cancer cell lines. These results are consistent with a regulatory role for RCAN1 in cancer cell motility in vitro. PMID- 19306111 TI - Heterogeneity of S-layer proteins from aggregating and non-aggregating Lactobacillus kefir strains. AB - Since the presence of S-layer protein conditioned the autoaggregation capacity of some strains of Lactobacillus kefir, S-layer proteins from aggregating and non aggregating L. kefir strains were characterized by immunochemical reactivity, MALDI-TOF spectrometry and glycosylation analysis. Two anti-S-layer monoclonal antibodies (Mab5F8 and Mab1F8) were produced; in an indirect enzyme-linked immunosorbent assay Mab1F8 recognized S-layer proteins from all L. kefir tested while Mab5F8 recognized only S-layer proteins from aggregating strains. Periodic Acid-Schiff staining of proteins after polyacrylamide gel electrophoresis under denaturing conditions revealed that all L. kefir S-layer proteins tested were glycosylated. Growth of bacteria in the presence of the N-glycosylation inhibitor tunicamycin suggested the presence of glycosydic chains O-linked to the protein backbone. MALDI-TOF peptide map fingerprint for S-layer proteins from 12 L. kefir strains showed very similar patterns for the aggregating strains, different from those for the non-aggregating ones. No positive match with other protein spectra in MSDB Database was found. Our results revealed a high heterogeneity among S layer proteins from different L. kefir strains but also suggested a correlation between the structure of these S-layer glycoproteins and the aggregation properties of whole bacterial cells. PMID- 19306112 TI - Pseudocercospora griseola causing angular leaf spot on Phaseolus vulgaris produces 1,8-dihydroxynaphthalene-melanin. AB - Pseudocercospora griseola is the causal agent of angular leaf spot of common bean (ALS). It has undergone parallel coevolution with its host and two major groups have been defined, "Andean" (P. griseola f. griseola) and "Mesoamerican" (P. griseola f. mesoamericana). The aim of this study was to analyze the nature and the level of the dark pigment synthesized by the representatives of each group. After 21 days of incubation on potato dextrose agar medium, P. griseola f. griseola isolate S3b developed colonies with diameters of 17.5 +/- 1.3 mm and concentric rings of pigmentation. Isolate T4 of P. griseola f. mesoamericana presented smaller colonies (9.9 +/- 0.3 mm) with a uniform dark-gray color. Both isolates, S3b and T4, produced the same pigment, a 1,8-dihydroxynaphthalene melanin, although different in quantity and structural features as suggested by the IR spectrum. The P. griseola f. griseola isolate S3b had a higher growth rate and melanin content as well as smaller sensitivity to melanin synthesis inhibitors compared to the isolate T4 of P. griseola f. mesoamericana. These results suggest a possible link between melanin and growth in P. griseola. PMID- 19306113 TI - HPLC-CHIP coupled to a triple quadrupole mass spectrometer for carbonic anhydrase II quantification in human serum. AB - A method for carbonic anhydrase II (CA II) absolute quantification in human serum is presented. This method is based on high-performance liquid chromatography (HPLC)-Chip microfluidic device incorporating a nanoelectrospray source interfaced to a triple quadrupole mass spectrometer. The fraction containing CA II was isolated by preparative reversed-phase HPLC, and peptides obtained from the tryptic digest of the protein mixture were separated by the HPLC-Chip system. The multiple-reaction monitoring acquisition mode of a selected suitable CA II peptide and peptide internal standard allowed the selective and sensitive determination of a CA II. Absolute recovery of the method was 52 +/- 12%, while analytical recovery was 81 +/- 10%. For the eight samples analyzed, the matrix effect was found to be only -14 +/- 6%. A comparison among three regression lines type which were obtained by external calibration, matrix-matched calibration, and standard addition method, respectively, demonstrated that the first one is adequate in obtaining good accuracy and precision. Method quantification limit for CA II in serum was estimated to be 2 fmol/mL. CA II mean concentration in sera from eight healthy subjects was found to be 56 pmol/mL (relative standard deviation 24%). PMID- 19306114 TI - A semi-quantitative dipstick assay for microcystin. AB - An immunochromatographic lateral flow dipstick assay for the fast detection of microcystin-LR was developed. Colloid gold particles with diameters of 40 nm were used as red-colored antibody labels for the visual detection of the antigen. The new dipstick sensor is capable of detecting down to 5 microg x l(-1) (ppb; total inversion of the color signal) or 1 ppb (observation of color grading) of microcystin-LR. The course of the labeling reaction was observed via spectrometric wave shifts caused by the change of particle size during the binding of antibodies. Different stabilizing reagents showed that especially bovine serum albumin (BSA) and casein increase the assays sensitivity and the conjugate stability. Performance of the dipsticks was quantified by pattern processing of capture zone CCD images. Storage stability of dipsticks and conjugate suspensions over 115 days under different conditions were monitored. The ready-to-use dipsticks were successfully tested with microcystin-LR-spiked samples of outdoor drinking- and salt water and applied to the tissue of microcystin-fed mussels. PMID- 19306115 TI - Disposable DNA biosensor with the carbon nanotubes-polyethyleneimine interface at a screen-printed carbon electrode for tests of DNA layer damage by quinazolines. AB - A screen-printed carbon working electrode within a commercially available screen printed three-electrode assembly was modified by using a composite of multiwalled carbon nanotubes (MWCNT) dispersed in polyethylenimine (PEI) followed by covering with the calf thymus dsDNA layer. Several electrochemical methods were used to characterize the biosensor and to evaluate damage to the surface-attached DNA: square wave voltammetry of the [Ru(bpy)(3)](2+) redox indicator and mediator of the guanine moiety oxidation, cyclic voltammetry and electrochemical impedance spectroscopy in the presence of the [Fe(CN)(6)](3-/4-) indicator in solution. Due to high electroconductivity and large surface area of MWCNT and positive charge of PEI, the MWCNT-PEI composite is an advantageous platform for the DNA immobilization by the polyelectrolyte complexation and its voltammetric and impedimetric detection. In this respect, the MWCNT-PEI interface exhibited better properties than the MWCNT-chitosan one reported from our laboratory previously. A deep DNA layer damage at incubation of the biosensor in quinazoline solution was found, which depends on the quinazoline concentration and incubation time. PMID- 19306116 TI - Archaeal diversity at the great salt plains of Oklahoma described by cultivation and molecular analyses. AB - The Great Salt Plains of Oklahoma is a natural inland terrestrial hypersaline environment that forms evaporite crusts of mainly NaCl. Previous work described the bacterial community through the characterization of 105 isolates from 46 phylotypes. The current report describes the archaeal community through both microbial isolation and culture-independent techniques. Nineteen distinct archaea were isolated, and ten were characterized phenetically. Included were isolates phylogenetically related to Haloarcula, Haloferax, Halorubrum, Haloterrigena, and Natrinema. The isolates were aerobic, non-motile, Gram-negative organisms and exhibited little capacity for fermentation. All of the isolates were halophilic, with most requiring at least 15% salinity for growth, and all grew at 30% salinity. The isolates were mainly mesothermic and could grow at alkaline pH (8.5). A 16S rRNA gene library was generated by polymerase chain reaction amplification of direct soil DNA extracts, and 200 clones were sequenced and analyzed. At 99% and 94% sequence identity, 36 and 19 operational taxonomic units (OTUs) were detected, respectively, while 53 and 22 OTUs were estimated by Chao1, respectively. Coverage was relatively high (100% and 59% at 89% and 99% sequence identity, respectively), and the Shannon Index was 3.01 at 99% sequence identity, comparable to or somewhat lower than hypersaline habitats previously studied. Only sequences from Euryarchaeota in the Halobacteriales were detected, and the strength of matches to known sequences was generally low, most near 90% sequence identity. Large clusters were observed that are related to Haloarcula and Halorubrum. More than two-thirds of the sequences were in clusters that did not have close relatives reported in public databases. PMID- 19306117 TI - Constructivisms from a genetic point of view: a critical classification of current tendencies. AB - In this paper, we propose a critical classification of contemporary constructivist orientations. Our fundamental theoretical reference is the notion of genesis, understood as the construction of reality in a way that is neither relativist nor positivist-realist. We identify a nucleus of classic, genetic constructivism that revolves around the ideas of Baldwin, Piaget and Vygotsky and discuss two tendencies that distort the spirit of that nucleus: objectivism and subjectivism. Objectivism rules out the psychological, constructive activity of the subject, subordinating (or just reducing) it to objective structures either from nature (like genetic endowment or neural functioning), or from culture (like language or social practices). Subjectivism completely detaches the objectivity of knowledge from its construction on the part of the subject, reducing it to the mere product of individual interest, view, or irrationality. Thus, subjectivism is the non-constructive way to conceive the subject. Then, we attempt to show the dialectics that exists between these two tendencies and the scope of our criteria by analysing a representative (non exhaustive) group of authors who are defined as constructivists or who bring important elements to the debate about constructivism. PMID- 19306118 TI - When stereotypes become 'scientific' statements: dealing with gender issues. AB - The theoretical analysis by Watzlawik (Integrative Psychological & Behavioral Science 2009) demonstrates the scientific fragility of the constructs of masculinity and femininity based on the oversimplification and overlapping between three levels of analysis: group differences, inter-individual differences and intra-individual differences. Watzlawik presents fresh and relevant contributions in terms of methodological issues, especially about the construction of scientific generalizations. Here I focus on issues related to the transformation of stereotypes in statements about gender differences that claim to be 'scientific'--outlining the socio-political agendas of such statements. PMID- 19306119 TI - Development and characterization of CD22-targeted pegylated-liposomal doxorubicin (IL-PLD). AB - Non-Hodgkin's lymphoma (NHL) is the sixth most common cause of cancer deaths in the U.S. Most NHLs initially respond well to chemotherapy, but relapse is common and treatment is often limited due to the toxicity of chemotherapeutic agents. Pegylated-liposomal doxorubicin (PLD, Ben Venue Laboratories, Inc), a produces less myelotoxicity than non-liposomal (NL) doxorubicin. To further enhance efficacy and NHL targeting and to decrease toxicity, we conjugated an anti-CD22 monoclonal antibody (HB22.7) to the surface of PLD, thereby creating CD22 targeted immunoliposomal PLD (IL-PLD). HB22.7 was successfully conjugated to PLD and the resulting IL-PLD exhibits specific binding to CD22-expressing cells as assessed by immunofluorescence staining. IL-PLD exhibits more cytotoxicity than PLD in CD22 positive cell lines but does not increase killing of CD22 negative cells. The IC(50) of IL-PLD is 3.1 to 5.4 times lower than that of PLD in CD22+ cell lines while the IC(50) of IL-PLD is equal to that of PLD in CD22- cells. Furthermore, IL-PLD remained bound to the CD22+ cells after washing and continued to exert cytotoxic effects, while PLD and NL- doxorubicin could easily be washed from these cells. PMID- 19306120 TI - [Preface: genetics and psychiatry]. PMID- 19306121 TI - [Psychopathology and temperament in parents of adolescents with substance use disorders: a controlled study]. AB - OBJECTIVE: The aim of the present study was to investigate the characteristics of psychopathology and temperament in parents of adolescents with substance use disorders (SUD). We compared the psychiatric diagnoses and temperament characteristics of parents of adolescents with SUD with parents of adolescents without SUD. METHODS: Diagnostic interviews of the parents were conducted with 3 instruments: Structured Clinical Interview for DSM non-patients, Axis 1 Disorders, and Axis 2 Disorders (SCID-NP, SCID-I, and SCID-II). Temperament characteristics were evaluated with TEMPS-A (Temperament Evaluation of Memphis, Pisa, Paris, and San Diego Autoquestionnaire), Turkish version. RESULTS: Mood disorders (P = 0.010), and avoidant (P = 0.005) and paranoid (P = 0.011) personality were more frequent in mothers of adolescents with SUD than in the controls, whereas mood disorders (P = 0.011), SUD (P = 0.047), and paranoid personality (P = 0.008) were more frequent in the fathers of adolescents with SUD than in the controls. Depressive, cyclothymic, and anxious temperament scores (respectively P = 0.023, P = 0.00, and P = 0.003) were higher in mothers than in the controls, and depressive temperament (P = 0.013) scores were higher in fathers of adolescents with SUD than in the controls. CONCLUSION: DSM Axis I and Axis II disorders were more frequent in the parents of adolescents with SUD than in the parents of adolescents without SUD. The difference in temperament characteristics exists in both groups. Familial effects have significant clinical implications for SUD in adolescents, such as individual features and peer effect. PMID- 19306122 TI - [Temperamental characteristics of mothers of preschool children with separation anxiety disorder]. AB - OBJECTIVE: It is reported that there are many risk factors for the development of separation anxiety disorder (SAD) in children and adolescents. One of the most important factors is the mother-child relationship. It is thought that the temperamental characteristics of the mother have an important role to play in the quality of this relationship. The present study aimed to determine the temperamental characteristics of mothers whose preschool children were diagnosed with separation anxiety disorder. METHOD: The study included 60 mothers of children with separation anxiety disorder (diagnosed between 4 and 7 years of age) and 60 mothers of healthy children who were matched by sociodemographic factors with children with SAD. All cases were evaluated with a sociodemographic form, the Child Behavior Check List (CBCL) and The Temperament Evaluation of Memphis, Pisa, Paris and San-Diego Auto questionnaire (TEMPS-A). RESULTS: Mean age of the children (32 female, 28 male) was 5.12+/-.0.85 years. The children with an anxiety disorder had higher behavioral problem CBCL scores than the control group. The mothers of the children with anxiety disorders had higher depressive, cyclothymic, irritabl and anxious temperament scores than the control group mothers. CONCLUSION: The mothers of the children with separation anxiety disorder had higher scores on depressive, cyclothymic, irritability and anxious temperament scores than the control group mothers. We think that this study can be used to inform future community based, prospective studies. PMID- 19306123 TI - [Proton magnetic resonance spectroscopy in Asperger's syndrome: correlations with neuropsychological test scores]. AB - OBJECTIVES: It has been shown that autistic spectrum patients have impaired theory of mind (ToM) performance; however, no study has investigated the relationship between ToM performance and brain neurochemistry in these patients. The present study aimed to investigate the correlations between dorsolateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC) N-acetyl-aspartate (NAA)/choline (Cho), NAA/creatine (Cr), and Cho/Cr values based on 1H magnetic resonance spectroscopy and ToM tests. METHOD: The study sample included 13 adult, right-handed, Caucasian males with Asperger's syndrome (AS) (age range: 17-37 years) and 20 controls matched by age, gender, handedness, and Wechsler Adult Intelligence Scale, Revised (WAIS-R) full-scale IQ scores. RESULTS: AS cases had significantly lower ToM performance. DLPFC NAA/Cho levels were inversely correlated to ToM scores (r = -0.738, P = 0.004). On the other hand, ToM performance improved as DLPFC Cho/Cr increased (r = 0.656, P = 0.015). ACC MRS variables were not significantly correlated with ToM performance in the AS group. No significant correlation was observed between ACC or DLPFC MRS variables and ToM performance in the control group. DISCUSSION: Because NAA/Cho was inversely correlated with ToM performance and Cho/Cr was correlated with ToM performance, it can be suggested that the Cho level was related to better ToM test performance in the AS group. An increase in the Cho peak was associated with an increase in membrane breakdown or turnover. The Cho peak was also thought to reflect cellular density and astrocytosis. It is suggested that membrane turnover and astrocytosis might affect cognitive functioning. PMID- 19306124 TI - [Testing the probability of a model to predict suicide risk in high school and university students]. AB - OBJECTIVE: The aim of this study was to investigate the validity of a model proposed by Batigun and Sahin regarding suicide probability. METHOD: The sample was composed of 2343 students aged 15-25 years that were attending various high schools and universities. According to the proposed model, 2 risk groups were formed from this sample, according to their scores on the investigation variables (those that simultaneously received high scores 1 standard deviation above the mean on the Problem Solving Inventory, Multidimensional Anger Scale, and Impulsivity Scale). Two other risk groups were formed according to the criteria variable scores (suicide probability scores 1 standard deviation above and below the mean). A series of analyses were conducted to investigate the similarity between the model risk groups and criteria risk groups. RESULTS: The results reveal that the model had a 43.3% success rate for predicting those with high suicide probability, while the false negative rate was 0%. Discriminant analysis showed that the model correctly discriminated 90.2% of those with low suicide probability and 87.3% of those with high suicide probability. CONCLUSION: The results support the validity of the proposed model for selecting individuals with high suicide probability. In addition, the model can be used to offer these individuals certain preventive measures, such as problem solving, communication skills, and anger management training. PMID- 19306125 TI - [Restraint variables in a regional mental health hospital in Turkey]. AB - OBJECTIVE: To obtain a database to be used in the effort to reduce the use of restraint in mental health hospitals in Turkey, and to compare the data with results of similar studies. METHOD: The sociodemographic characteristics, history of violent or suicidal behavior, and BPRS scores of patients restrained during a 1-month period, and data concerning the number of beds and staff working during day and night shifts were collected. The restraint variables, number of restraints used and duration of restraint use, in all wards were determined and adult psychosis wards were compared according to these variables. RESULTS: Approximately 25% (194) of 810 hospitalized patients were restrained for a total of 1524.25 h (mean: 3.25 h) during 1 month. In all, 182 restrained patients were analyzed and most were male, elementary school graduates, never married, and living with first-degree relatives. There were no correlations between these variables, and the number of restraints used and the duration of their use. Mean age of the females (36.27 years) was higher than that of the males (32.46 years); the difference was significant. In all, 66.6% of restraint use was because of violent behavior or the probability of violent behavior. The rate of restraint use was lower during day shifts (43.82%). Patients on adult wards were not different in terms of diagnosis, BPRS score, rate of violent behavior, and reasons for using restraint, but were markedly different in terms of the number of restraints used and the duration of restraint. CONCLUSION: The number of restraints used and the duration of restraint was on average level. Restraint was associated with being female and older, and having more severe symptoms. Night and weekend shifts were more risky for amount and duration of restraint. Non clinical institutional factors were a determining factor in the use of restraint. PMID- 19306126 TI - [Demographic and clinical correlates of length of stay in a Nigerian university hospital psychiatric unit]. AB - OBJECTIVE: To investigate the clinical and demographic correlates of prolonged hospital stays in a cohort of Nigerian psychiatric in-patients. METHODS: The investigation covered a 5 year period of psychiatric inpatient admissions at a Nigerian general hospital psychiatric and the records of 584 admitted and discharged patients that fulfilled the inclusion criteria were analyzed. Data were collected on demographic and clinical characteristics of the patients. Univariate and multivariate analyses were conducted to identify factors associated with length of hospital stay. RESULTS: Mean length of stay (LOS) was 25 +/- 23 days. Only the clinical variables of a primary diagnosis of schizophrenia (P=0.0001), having received ECT (P=0.0001), and a history of previous hospitalization for psychiatric illness (P=0.034) significantly predicted prolonged LOS. CONCLUSION: As cost considerations are very important in this time of evidence-based practice there is a need for clinicians to modify their practice and be aware of the patient-related variables that affect LOS when managing their clients. PMID- 19306127 TI - [The development of the life evaluation questionnaire for hemodialysis patients]. AB - OBJECTIVE: Existing quality of life measures fail to incorporate patients' own understanding of evaluation of life following the diagnosis of end stage renal failure (ESRF). Previous qualitative research has identified ways of evaluating life that have not been captured by existing quantitative research and that can be possible targets for clinical practice. Nonetheless, quantification is necessary if clinicians are to be informed of potentially important ways of evaluating life. It is also necessary to devise patient-derived measures of quality of life if the aim is also to monitor improvement in ways of evaluating life following interventions devised to change them. METHOD: Three studies were carried out. In the first two, the ways of evaluating life that were previously identified were quantified, a questionnaire that measures ways of evaluating life was developed, and the clinical utility of this questionnaire was examined in a sample of 156 hemodialysis patients. In the third study the predictive validity of this questionnaire was examined in a sample of 153 hemodialysis patients. RESULTS: The findings identified a number of targets for clinical practice. These included feeling unable to 'get round' limitations, mixed negative feelings about caregivers, and failure to find something positive about ESRF The newly developed 3 subscales of life evaluation were internally consistent. The findings also show that thes subscales had construct and predictive validity. CONCLUSION: The present studies provide a disease-specific life evaluation questionnaire consisting of 3 subscale for use in hemodialysis patients that can supplement existing generic quality of life measures. PMID- 19306128 TI - [The reliability and validity of Turkish Brief Measure of Worry Severity based on Turkish university students]. AB - OBJECTIVE: The aim of this study was to examine reliability and validity of the Turkish Brief Measure of Worry Severity (BMWS) for assessing the level of dysfunctional worry. METHOD: The study sample consisted of two different student groups from various faculties of Hacettepe University (age range: 17-25 years). First, data were collected from the group, composed of 210 female and 170 male students to evaluate the scale's test re-test correlation, Cronbach's alpha coefficient, and criterion and convergent validity. Next, the Beck Depression Inventory (BDI) and Beck Anxiety Inventory (BAI) were administered to the other group, composed of 805 students to assess distinctive validity of the Turkish BMWS. Students that scored over a quarter on median of BDI were regarded as having severe depression symptoms and the students that scored > 26% on BAI were regarded as having severe anxiety symptoms. In this manner anxiety (12 male, 38 female) and depression groups (4 male, 10 female) were formed. RESULTS: Following a series of analyses, test-retest correlation (r = 0.76) and internal consistency (Cronbach's alpha 0.88) of the Turkish BMWS was determined to be statistically high. While the Turkish BMWS showed unifactorial construct, the scores of the scale differentiated between the depression group and the anxiety group (depression group: X = 8.14, SS = 4.03; anxiety group: X = 11.56, SS = 5.11). Furthermore, the Turkish BMWS's correlations with the Name Penn State Worry Questionnaire (r = 0.75) and Trait State Anxiety Inventory (State Anxiety Inventory: r= 0.42; Trait Anxiety Inventory r = 0.72) were statistically significant and supported the scale's construct validity. CONCLUSION: The results show that the Turkish BMWS is a reliable and valid scale for use with Turkish university students. PMID- 19306129 TI - [Assessment of early maladaptive schemas: a psychometric study of the Turkish young schema questionnaire-short form-3]. AB - OBJECTIVE: Schemas are generally assumed to develop as a result of early experiences with attachment figures. Within this theoretical framework Young et al. (1991, 2003) developed a schema questionnaire to evaluate early maladaptive schemas. The aim of the present study was to preliminarily establish the psychometric validity and reliability of the Turkish version of the Young Schema Questionnaire-Short Form-3. METHOD: University students from different departments participated in the study. According to phases of psychometric examinations, the number of participants differed in a range of N= 150-1071. RESULTS: Principal components analysis with equamax rotation was carried out and 14 definable factor structures emerged. Higher-order factor analysis supported 5 schema domains: Impaired autonomy, disconnection, unrelenting standards, other directedness, and impaired limits. Test-retest and internal consistency analysis revealed statistically significant correlation coefficients, which can be interpreted as evidence of the reliability. As to convergent validity, correlational analysis of theoretically-related variables (SCL-90-R) showed statistically significant coefficients and the direction of the relationships were congruent with theoretical expectations. Furthermore, in a pilot study, we examined the discriminant validity of the scale. Accordingly, t-test analysis that compared the YSQ-subscale scores of clinical and normal populations yielded statistically significant differences in some schemas and schema domains. CONCLUSION: As preliminary evidence, our findings show that the factor structure of the Turkish YSQ-SF3 is generally consistent with previous studies and that it has acceptable levels of reliability and validity. PMID- 19306130 TI - [The genetics of suicidal behavior]. AB - There is consistent evidence suggesting that genetic factors play an important role in predisposition to suicidal behavior. Family, twin, and adoption studies have demonstrated that there is a genetic dimension to suicide. Although there is some overlap between suicide and mood disorders, even among psychiatric groups with the highest risk, some patients never attempt suicide, indicating the importance of a diathesis or genetic link to suicide that is independent of the underlying psychiatric disorder. Over the last 3 decades research has shown that there is a relationship between suicide, aggressiveness, and impulsivity. It is possible that genetic factors may be related to personality traits such as impulsiveness and aggressiveness, which in turn may lead to suicide attempts. An increasing number of molecular genetic studies have been carried out among cases involving suicidal behavior and candidate genes thought to be related to suicide. The most important candidate genes include the serotonin transporters (SERTs), tryptophan hydroxylase (TPH), some serotonin receptors (5HT1A, 5HT1B, and 5HT2A), catechol-O-methyltransferase (COMT), monoamine oxidase-A (MAO-A), and tyrosine hydroxylase (TH). The aim of this review was to assess the genetic dimension of suicide behavior, both current and historical. PMID- 19306131 TI - [The role of low-dose pramipexole in the treatment of treatment-resistant bipolar depression: a case report]. AB - Despite a wide range of various drugs, a significant proportion of depressed bipolar patients fail to respond to the treatment strategies. Novel theraupetics for bipolar depression are needed. Preliminary studies suggest that pramipexole a dopaminergic agent that has been used in the treatment of Parkinson's disease and restless leg syndrome may have antidepressant properties in unipolar and bipolar depressed patients as well as neurotrophic properties. The optimal antidepressant daily dose of pramipexole is not known. It has been suggested to be used between 0.125 to 9.0 mg/day. In double blind placebo controlled bipolar depression treatment studies, the average daily dose of pramipexole was 1.7 mg. Manic switches have been reported with depressive subjects and with subjects without any mental disorders. We report two cases of treatment resistant bipolar depression. Despite different treatment strategies and treatment adherence, the patients did not give optimal response to the treatments and continue to experience depressive relapses. They have been treated with low dose (0.5-0.75 mg/day) pramipexole augmentation successfully. The severity and the duration of the depressive episodes were decreased. No serious adverse event has been reported with pramipexole during the maintenance treatment. PMID- 19306132 TI - Predictors of stillbirth among HIV-infected Tanzanian women. AB - OBJECTIVE: To determine maternal risk factors for stillbirth among pregnant HIV infected women in sub-Saharan Africa. DESIGN: Prospective cohort study nested within a micronutrient trial. At enrollment, maternal sociodemographic, obstetric, immunologic, clinical, and nutritional variables were measured. Women were followed through monthly clinic visits until delivery. Multivariate predictors of stillbirth were identified in Poisson regression models. SETTING: Antenatal clinic in a tertiary care hospital in urban Dar es Salaam, Tanzania. POPULATION: N=1,078 women enrolled between 12 and 27 weeks of gestation. MAIN OUTCOME MEASURES: Stillbirth (delivery of dead baby > or = 28 weeks' gestation), fresh stillbirth, and macerated stillbirth. RESULTS: Among 1,017 singleton pregnancies, there were 49 stillbirths, yielding a stillbirth risk of 50.0 per 1,000 deliveries (95% Confidence Interval(CI) = 37.2, 65.6). Of stillbirths with known type, 53.7% were fresh and 46.3% macerated. In multivariate analyses, baseline measures of late (> or = 21 weeks' gestation) study entry (Relative Risk (RR) = 2.13, 95% CI = 1.17, 3.87), CD3 count > or = 1,179 cells/ml (RR = 2.15, 95% CI = 1.16, 4.01), stillbirth history (RR = 3.53, 95% CI = 1.30, 9.59), primiparity (RR = 3.65, 95% CI = 1.83, 7.29), and syphilis infection (RR = 2.06, 95% CI = 1.09, 3.88) predicted increased stillbirth risk. Late study entry, illiteracy, stillbirth history, primiparity, CD3 count > or = 1,179 cells/ml, gonorrhea infection, and previous hospitalization predicted increased risk of fresh stillbirth, while living alone and syphilis infection predicted increased risk of macerated stillbirth. CONCLUSIONS: Applying antenatal screening and preventive tools for the socioeconomic, obstetric, immunologic, and clinical risk factors identified may assist in reducing the high incidence of stillbirth among HIV-infected women in urban sub-Saharan Africa. PMID- 19306133 TI - Correlation between Aurora-A expression and the prognosis of cervical carcinoma patients. AB - OBJECTIVE: Aurora-A, a novel member of the serine/threonine kinase family, has been reported to be correlated with tumorigenesis. Our aim was to investigate whether Aurora-A expression correlates with clinicopathologic factors and prognosis of cervical carcinoma patients. DESIGN/SETTING: Retrospective study. POPULATION: Seventy-four cervical carcinoma patients, between 1996 and 2002. METHODS: Reverse transcription-polymerase chain reaction and Western blot assays were performed to detect the expression of Aurora-A gene in cervical carcinoma cells and paired cancerous and corresponding noncancerous tissues from 74 cervical carcinoma patients. The expression of Aurora-A protein in tissues was also determined by immunohistochemistry. The relationships of Aurora-A expression with clinical factors and prognosis of patients were evaluated by statistical analysis. RESULTS: The expression of Aurora-A mRNA and protein was significantly higher in cervical carcinoma cells than in normal cervical epithelial cell (p<0.05). The expression of Aurora-A mRNA in cancerous tissues was significantly higher than that in corresponding noncancerous tissues (p<0.001). The expression of Aurora-A protein was also increased in tumor tissues by immunochemistry. Aurora-A transcript expression was correlated with FIGO stage (p=0.018), tumor differentiation (p=0.014), parametrial invasion (p=0.024), lymphnode or hematogenous metastasis (p=0.005 or 0.019), but not other clinicopathological factors. Patients with high Aurora-A expression had a poorer disease-free survival and overall survival rates than patients with low Aurora-A expression. Multivariate analysis showed that high Aurora-A mRNA expression was an independent prognostic factor (risk ratio: 2.88; p=0.005). CONCLUSIONS: Aurora-A might be used as a prognostic marker for cervical carcinoma patients. PMID- 19306134 TI - Jordanian obstetricians' personal preference regarding mode of delivery. AB - The personal preference of Jordanian obstetricians regarding mode of delivery in uncomplicated pregnancy with singleton cephalic presentation at term was surveyed by an anonymous structured questionnaire distributed at five conferences/scientific meetings on obstetrics and gynecology held in Jordan in 2007. The response rate was 70% (n=315), and 22 (7%) of the respondents chose elective cesarean section (CS). Respondents <45 years seemed to be more in favor of elective CS than those >or=55 years (14.1% vs. 3.7%). In multivariate analysis, increased age was inversely associated with the odds of choosing elective CS (OR = 0.93, 95% CI: 0.88-0.99, p=0.024) after adjusting for gender, sector, and duration of practice. The main reasons for preferring abdominal delivery were the fear of long-term sequel (stress incontinence and anal sphincter damage) and the wish to preserve sexual function (86.4 and 50.0%, respectively). The majority of Jordanian obstetricians and gynecologists preferred vaginal delivery in uncomplicated pregnancy for themselves or their wives. PMID- 19306135 TI - Efficacy of metformin supplementation during ovarian stimulation of lean PCOS patients undergoing in vitro fertilization. AB - OBJECTIVE: To evaluate the effect of metformin addition during ovarian hyperstimulation on cycle parameters and outcome of in vitro fertilization (IVF) treatment in polycystic ovary syndrome (PCOS) patients with a body mass index (BMI) <28 kg/m(2). DESIGN: Retrospective review of patients' records. SETTING: Istanbul Memorial Hospital Assisted Reproductive Treatment Unit. POPULATION: A total of 339 non-obese PCOS patients undergoing IVF were evaluated according to the supplementation of metformin. METHODS: Cycle parameters and IVF outcomes of 220 patients given metformin were compared to those of 119 patients treated without metformin. MAIN OUTCOME MEASURES: Implantation and pregnancy rates. RESULTS: Metformin co-treatment led to significantly lower peak E2 levels (3,481 pg/ml vs. 4,192 pg/ml; p <0.0001). However, gonadotropin consumption, stimulation duration, numbers of total and mature oocytes retrieved, fertilization rate, and ratio of grade I embryos developed were similar in both groups. In the metformin administered group, significantly higher implantation (25% vs. 18%; p=0.003) and pregnancy rates (58% vs. 45%; p=0.04) were achieved. Abortion and moderate-severe ovarian hyperstimulation syndrome rates were found to be similar in both groups. CONCLUSIONS: Although metformin use was especially and strongly recommended in obese PCOS cases, its supplementation during IVF in PCOS patients with a BMI <28 kg/m(2) was observed to be beneficial and efficacious. PMID- 19306136 TI - Penetration of moxifloxacin into rat mandibular bone and soft tissue. AB - OBJECTIVE: Based on its in vitro activity and spectrum of activity, the new 8 methoxyquinolone antibiotic moxifloxacin (MXF) seems suited for the antibiotic therapy of odontogenic infections. Penetration into the relevant tissue is another prerequisite for clinical efficacy. For this reason, the levels of MXF in plasma, soft tissue, and mandibular bone were determined in an animal model with Wistar rats. MATERIAL AND METHODS: Samples of 49 rats were analyzed. Tissue samples were homogenized and proteins were precipitated. The pharmacokinetic evaluation was conducted based on non-compartmental analysis. RESULTS: The concentration-time courses of tissues show a more plateau-shaped curve compared to plasma. Calculated AUC (area under the curve) ratios tissue:plasma were M. masseter:plasma = 2.64 and mandibles:plasma = 1.13. CONCLUSIONS: Administration of antibiotics is considered an important part of therapy during and/or after surgical procedures in the maxillofacial area. Because of the good penetration into bone and muscle tissues demonstrated in Wistar rats, MXF might be an option for clinical application in this indication. PMID- 19306137 TI - Composite material stent comprising metallic wire and polylactic acid fibers, and its mechanical strength and retrievability. AB - BACKGROUND: Although metallic stents are characterized by strong expanse of force, thin walls, and easy stent deployment, their removal from the body is usually difficult or impossible due to the difficulty of unraveling their mesh structure. A stent built of a composite material comprising a metallic wire and a polylactic acid (PLA) fiber, in which the metallic wire component could be unraveled after PLA fiber degradation in the body, should allow easy stent removal. PURPOSE: To evaluate the mechanical strength and retrievability of a composite material stent comprising a metallic wire and a PLA fiber. MATERIAL AND METHODS: We produced a composite material stent comprising a metallic wire and a biodegradable fiber (hybrid stent). As the metallic wire is not cross-linked with itself, but with the PLA fibers only, the hybrid stent can be easily unraveled after PLA fiber degradation. This stent was built with a 0.2-mm stainless-steel wire and a 0.23-mm PLA fiber knitted in the same textile as an Ultraflex stent. For comparison, an identical stent was built using PLA fiber only (PLA stent). The mechanical strength of these stents was tested by the radial expansive force response against circumferential shrinkage stress load. Change in radial force due to PLA fiber degradation was estimated by adding an artificial PLA degeneration process, by immersing each stent in a water bath at 80 degrees C for 48 hours. Retrievability of the hybrid stent after PLA degeneration was examined by hooking and pulling out the residual stainless-steel wire from a silicon tube. RESULTS: The hybrid stent exhibited a linear response in radial expansive force within the range of 15% diameter reduction. The PLA stent did not exhibit linear response at over 15% diameter reduction. Decrease of radial expansive force after PLA degradation was within 5% of the original force in the hybrid stent, but the PLA stent did not create effective radial expansive force. Hybrid stents, even after PLA degradation, exhibited a linear response in radial expansive force, within the range of 15% diameter reduction. The metallic component of the heat processed hybrid stent was easily unraveled by pulling out the wire. CONCLUSION: The hybrid stent comprising a stainless-steel wire and a PLA fiber appears to provide effective radial expansive force and retrievability. PMID- 19306138 TI - Involvement of the proximal tibiofibular joint in ankylosing spondylitis. AB - BACKGROUND: Ankylosing spondylitis (AS) may affect peripheral joints, with the shoulder, hip, and knee being well known involved sites. However, involvement of the proximal tibiofibular (PTF) joint has not yet been investigated. PURPOSE: To evaluate PTF joint abnormalities in patients with AS. MATERIAL AND METHODS: From July 1997 to June 2005, 16 patients (15 male, one female; mean age 25 years), who were clinically diagnosed with AS, underwent magnetic resonance imaging (MRI) to evaluate knee pain. All patients also underwent plain radiographs of the knee, lumbar spine, and pelvis. Twenty knee MRIs (bilateral in four patients) and 16 sets of knee, lumbar spine, and pelvic radiographs were retrospectively reviewed in order to evaluate possible AS involvement. The presence of abnormalities suggesting AS involvement were recorded separately in the sacroiliac joints, lumbar spine, hip, and femorotibial and PTF joints. If the PTF joint showed any pathologic findings, the radiologic findings were recorded. RESULTS: Three of 16 patients (18.7%) had pathologic features of the PTF joint observed by plain radiographs or MRI. One of these three patients showed bilateral involvement of the PTF joints on plain radiographs, while the other two patients showed unilateral involvement on MRI. Subchondral sclerosis, cartilage abnormality, erosion, and abnormal bone marrow signal intensity were identified on MRI. Plain radiographs of two patients revealed subchondral sclerosis and spur formation in the PTF joint. The frequencies of involvement of other joints in the 16 patients were as follows: lumbar spine, n=5 (31%), hip joint, n=1 (6%) (identified by plain radiographs), and femorotibial joints, n=10 (62.5%) (identified by knee MRI). CONCLUSION: MR imaging of the PTF joint can depict synovial changes and their effect on joint structures in patients with AS. The MRI findings of AS involving the PTF joints are subchondral sclerosis, cartilage abnormality, erosion, and abnormal bone marrow signal intensity. PMID- 19306140 TI - Aquaporin-4 as a potential marker of BBB disruption in ALS models. PMID- 19306141 TI - TDP-43 in skeletal muscle of patients affected with amyotrophic lateral sclerosis. AB - TAR DNA binding protein (TDP-43) is the pathologic substrate of neuronal and glial aggregates in amyotrophic lateral sclerosis (ALS). Pathologic TDP-43 is hyperphosphorylated and cleaved to generate abnormal protein species that accumulate in the cytoplasm. To assess the hypothesis of TDP-43 pathology as a systemic disorder in ALS we analysed the immunohistochemical and biochemical profile of TDP-43 in muscle biopsies of 30 ALS patients and 30 controls. In all ALS muscle biopsies we observed that TDP-43 was constantly present in an intranuclear localization and TDP-43 Western blotting showed only a 43-KDa band as controls. Our results suggest that TDP-43 pathology is probably confined to the central nervous system in ALS. PMID- 19306143 TI - Effects of initial surface wettability on biofilm formation and subsequent settlement of Hydroides elegans. AB - Hydroides elegans is a major fouling organism in tropical waters around the world, including Pearl Harbor, Hawaii. To determine the importance of initial surface characteristics on biofilm community composition and subsequent colonization by larvae of H. elegans, the settlement and recruitment of larvae to biofilmed surfaces with six different initial surface wettabilities were tested in Pearl Harbor. Biofilm community composition, as determined by a combined approach of denaturing gradient gel electrophoresis and fluorescence in situ hybridization, was similar across all surfaces, regardless of initial wettability, and all surfaces had distinct temporal shifts in community structure over a 10 day period. Larvae settled and recruited in higher numbers to surfaces with medium to low wettability in both May and August, and also to slides with high wettability in August. Pearl Harbor biofilm communities developed similarly on a range of surface wettabilities, and after 10 days in Pearl Harbor all surfaces were equally attractive to larvae of Hydroides elegans, regardless of initial surface properties. PMID- 19306144 TI - Role of type 1 fimbriae and mannose in the development of Escherichia coli K12 biofilm: from initial cell adhesion to biofilm formation. AB - The influence of type 1 fimbriae, mannose-sensitive structures, on biofilm development and maturation has been examined by the use of three isogenic Escherichia coli K12 strains: wild type, fimbriated, and non-fimbriated. Experiments with the three strains were done in minimal medium or Luria-Bertani broth supplemented with different concentrations of d-mannose. The investigation consisted of: (1) characterizing the bacterial surface of the three strains with respect to hydrophilicity and surface charge, (2) investigating the effect of type 1 fimbriae on bacterial adhesion rate and reversibility of initial adhesion on glass surfaces, and (3) verifying the role of type 1 fimbriae and exopolysaccharides (EPS) in biofilm maturation. The results suggest that type 1 fimbriae are not required for the initial bacterial adhesion on glass surfaces as the non-fimbriated cells had higher adhesion rates and irreversible deposition. Type 1 fimbriae, however, are critical for subsequent biofilm development. It was hypothesized that in the biofilm maturation step, the cells synthesize mannose rich EPS, which functions as a 'conditioning film' that can be recognized by the type 1 fimbriae. PMID- 19306145 TI - Mini-review: quorum sensing in the marine environment and its relationship to biofouling. AB - Bacterial quorum sensing (QS) is a cell-cell communication and gene regulatory mechanism that allows bacteria to coordinate swarming, biofilm formation, stress resistance, and production of toxins and secondary metabolites in response to threshold concentrations of QS signals that accumulate within a diffusion-limited environment. This review focuses on the role of QS signaling and QS inhibition in marine bacteria by compounds derived from marine organisms. Since the formation of a biofilm is considered to be an initial step in the development of fouling, direct and indirect effects of QS signals and inhibitors on the process of marine biofouling are discussed. Directions for future investigations and QS-related biotechnological applications are highlighted. PMID- 19306146 TI - Drinking motives as mediators of social anxiety and hazardous drinking among college students. AB - Although social anxiety (SA) and alcohol use disorders commonly co-occur, the relationship between these variables in college populations has been inconsistent. The present study tested the hypothesized model that negatively reinforcing, but not positively reinforcing, drinking motives (or reasons for drinking) would mediate the association between SA and three aspects of hazardous drinking (quantity/frequency, consequences, and dependence symptoms) in an ethnically diverse sample of college drinkers (N = 817; mean age = 19.9 years, range = 18-29). Structural equation modeling (SEM) results using the asymmetrical distribution of products test indicated that coping motives partially mediated the relationship between SA and negative consequences and dependence symptoms but not the quantity/frequency outcome. Contrary to the hypothesized model, conformity motives did not mediate the association between SA and hazardous drinking. As expected, positive reinforcement motives did not mediate the SA hazardous drinking association. Multigroup SEM analyses revealed that the mediation models did not differ for men (n = 215) and women (n = 602). Overall, the present findings support extant research and theoretical models regarding the mediating role of coping motives in the relationship between SA and problem drinking, suggesting a potential pathway for the development and maintenance of SA and alcohol use disorder comorbidity. Such findings could contribute to improved intervention programs by targeting coping drinking motives and building coping skills. PMID- 19306147 TI - The acceptability of computer-aided cognitive behavioural therapy: a pragmatic study. AB - The clinical and cost-effectiveness of a computer-aided cognitive behavioural therapy (CCBT) programme, Beating the Blues, is indicated by a number of studies, but relatively little is known about its acceptability for patients with depression, anxiety, or both. This study investigated the acceptability of Beating the Blues offered on eight scheduled clinic visits with brief face-to face support. Pre and posttreatment measures explored the relationship among programme acceptability, treatment continuation, and outcomes for people accessing the programme in routine care. Two hundred and nineteen patients with depression, anxiety, or both were offered Beating the Blues in 11 primary and secondary care practices. One hundred and ninety-one (87%) completed the pretreatment measures and 84 (38%) completed a treatment feedback questionnaire. Analysis of treatment acceptability for CCBT indicated a positive patient experience with the programme. Pretreatment expectancies predicted CCBT treatment completion but not outcomes. No differences were found between men and women on pretreatment measures. Posttreatment, women reported more favourable responses to the therapy, finding the programme more helpful and more satisfactory, than did men. No relationship between treatment acceptability and age was found. Study limitations, including research methods and attrition rates, and implications for future research are discussed. It is concluded that the Beating the Blues CCBT programme is an acceptable treatment for common mental health problems in routine care. PMID- 19306148 TI - Computer-aided self-exposure therapy for phobia/panic disorder: a pilot economic evaluation. AB - Phobia/panic disorder is common. It improves with exposure therapy, even when guided mainly by a computer self-help system such as FearFighter (FF), but such therapy must also demonstrate cost-effectiveness. This study compares the cost effectiveness of FF with computed-aided relaxation and clinician-led exposure. Data were obtained on patients from a randomised controlled trial of FF. Economic analyses used pretreatment and 1-month follow-up self-ratings of the main problem and global phobia. Clinician costs were calculated using the number of therapist hours and the cost of FF. Incremental cost-effectiveness ratios were calculated and cost-effectiveness acceptability curves were produced. Data were available on 62 patients with main problem ratings and 60 with global phobia ratings. FF and clinician-led exposure were more effective than relaxation but more expensive. Compared with relaxation, producing an extra unit of improvement on the main problem scale cost 64 UK pounds with FF and 100 UK pounds with clinician-led exposure. FF appeared to be more cost-effective using the global phobia rating (112 UK pounds per extra unit of improvement vs. 128 UK pounds for clinician-led exposure). The cost-effectiveness of FF could be enhanced if users had less highly trained supporters. FF would be less cost-effective if face-to-face therapy was delivered by less qualified professionals. Caution is urged regarding these indicative findings given that these were secondary analyses. PMID- 19306149 TI - Internet-based treatment for panic disorder: does frequency of therapist contact make a difference? AB - Internet-based interventions with therapist support have proven effective for treating a range of mental health conditions. This study examined whether frequency of therapist contact affected treatment outcomes. Fifty-seven people with panic disorder (including 32 with agoraphobia) were randomly allocated to an 8-week Internet-based cognitive behavioural treatment intervention (Panic Online) with either frequent (three e-mails per week) or infrequent (one e-mail per week) support from a psychologist. Posttreatment, intention-to-treat analyses revealed that both treatments were effective at improving panic disorder and agoraphobia severity ratings, panic-related cognitions, negative affect, and psychological and physical quality of life domains, with no differences between conditions. High end-state functioning was achieved by 28.6% of the frequent and infrequent participants, respectively. Therapist alliance, treatment credibility, and satisfaction also did not differ between groups, despite significantly greater therapist time invested in the frequent contact condition. The results provide evidence that the effectiveness of Internet-based mental health interventions may be independent of the frequency of therapist support and may, therefore, be more cost-effective than previously reported. PMID- 19306150 TI - Contested innocence-sexual agency in public and private space. Abstracts of the International Association for the Study of Sexuality, Culture and Society (IASSCS) Conference 2009. April 15-18, 2009. Hanoi, Viet Nam. PMID- 19306152 TI - Stabilization of overweight prevalence in French children between 2000 and 2007. AB - OBJECTIVE: Over the past several decades, the prevalence of childhood obesity has increased worldwide. In France, the National Nutrition and Health Program (PNNS), implemented in 2001, was aimed at stopping this increase. The purpose of our study was to monitor changes in prevalence of overweight and obesity in 7-9-year old children in France since the PNNS was set up. METHODS: A cross-sectional study was conducted in 2007 in France based on a comparable protocol used in 2000. Primary schools were randomly selected in 11 out of 26 administrative school districts. In each of these schools, two classes were randomly selected. All children from these classes were included. Weight and height were measured by nurses or physicians belonging to the French educational system. A lifestyle self administered questionnaire was completed by the parents. International Obesity Task Force cut-offs were used to define body mass status. Direct standardization was used to enable a comparison between 2000 and 2007. RESULTS: In 2007, the prevalence of overweight was 15.8%, including 2.8% obesity, compared with 18.1% and 3.8%, respectively, in 2000. Differences between 2000 and 2007 did not reach statistical significance. In both years, the risk of being overweight or obese was significantly lower for children whose parents' occupation was that of manager or white collar rather than the other, less-privileged occupational categories. CONCLUSION: Our study demonstrates stabilization of the prevalence of overweight and obesity in 7-9-year-old children in France between 2000 and 2007. It is possible that the PNNS may be partly responsible for this stabilization. PMID- 19306153 TI - Inclusion phenomena of clove oil with alpha-, beta-, gamma- and heptakis (2,6-di O-methyl)-beta-cyclodextrin. AB - Inclusion interactions of alpha-, beta-, gamma- and heptakis (2,6-di-O-methyl) beta-cyclodextrin (DMbeta-CD) as hosts with clove oil (an impure eugenol, I-Eug) as guest in aqueous solution were investigated by fluorescence emission spectra. The binding constants of different hosts to I-Eug in aqueous solution decreased in the order: gamma- > beta- > DMbeta- > alpha-CD. Two solid supramolecular inclusion complexes, I-Eug-beta-CD and I-Eug-gamma-CD, were prepared and characterised by nuclear magnetic resonance, powder X-ray diffraction, infrared spectroscopy, and thermogravimetric analysis. All the results proved the formation of I-Eug-CD. The inclusion differences between I-Eug and pure eugenol were discussed. The relative contents of the main component eugenol (Eug), second component (eugenol acetate, Eua) and others in I-Eug were found to be fairly different before and after being included by beta-CD, according to the data obtained from high performance liquid chromatography. This could be a practical method to extract the effective components (Eug and Eua) from I-Eug. PMID- 19306154 TI - The prevalence of seasonal affective disorder (SAD) in Greenland is related to latitude. AB - The aim of this study was to determine the prevalence of seasonal affective disorder (SAD) in Greenlanders and Danes living at four different latitudes in Greenland. A Seasonal Pattern Assessment Questionnaire (SPAQ) was mailed to 6021 men and women between the ages of 18 and 59 years living in four different municipalities in Greenland. The recipients were randomly selected from the National Population Register. Approximately 9% of the respondents met the criteria for SAD, and the incidence of SAD varied between a southern municipality and three northern municipalities. The prevalence of SAD was particularly high in northern municipalities. No significant difference was found in the prevalence of SAD between Greenlanders and Danes. The results are comparable with other population studies that have reported a high prevalence of SAD in arctic areas. The clinical implications of our findings and the possibilities for introducing light therapy should be assessed in future studies. PMID- 19306155 TI - Penrose updated: deinstitutionalization of the mentally ill is not the reason for the increase in violent crime. PMID- 19306156 TI - A symptom self-rating scale for schizophrenia (4S): psychometric properties, reliability and validity. AB - The objective of the study was to validate a self-administrated symptom rating scale for use in patients with schizophrenia spectrum disorders by item analysis, exploration of factor structure, and analyses of reliability and validity. Data on 151 patients, initially treated by risperidone, obtained within the framework of a naturalistic Phase IV longitudinal study, were analysed by comparing patient and clinician ratings of symptoms, side-effects and global indices of illness. The Symptom Self-rating Scale for Schizophrenia (4S) is psychometrically adequate (item analysis, internal consistency, factor structure). Side-effect ratings were reliable. Symptom ratings displayed consistent associations with clinicians' ratings of corresponding symptom dimensions, suggesting construct validity. Patients had most difficulties assessing negative symptom items. Patients were well able to assess their own symptoms and drug side-effects. The factor structure of symptom ratings differs between patients and clinicians as well as how they construe global indices of illness. Clinicians focus on psychotic, patients on affective symptoms. Use of symptom self-ratings is one way to improve communication and thereby strengthen the therapeutic alliance and increase treatment adherence. PMID- 19306157 TI - Acute bacterial meningitis in adults: predictors of outcome. AB - Case fatality ratio and permanent sequelae of acute bacterial meningitis remain high in recent decades. A prospective longitudinal study of adult patients admitted with community acquired acute bacterial meningitis at a tertiary infectious diseases unit aimed to identify predictors of unfavourable outcome - death and sequelae. Anamnestic, clinical and laboratory data and clinical outcome were recorded. From 1997 to 2006, 279 adults (122F, 157M) with a median age of 51 y were admitted with acute bacterial meningitis. Predisposing condition and comorbidity were recorded in 42% and 38% of patients, respectively. Time between symptoms onset and antibiotic treatment ranged from 6 to 160 h. An aetiological agent was identified in 77% of patients: Streptococcus pneumoniae (29%) and Neisseria meningitidis (27%) were the most frequent. 55 patients (20%) died and 63 (23%) had neurological sequelae 6 months after discharge. In multivariate analysis, 7 independent predictors of unfavourable outcome were identified: internal comorbidity, time to treatment >48 h, coma, hypotension, high CSF protein, low glucose ratio, and non-meningococcal aetiology. The results suggest that acute bacterial meningitis remains associated with significant morbidity and mortality. Maintaining a high clinical suspicion and initiating appropriate diagnostic testing and therapeutic interventions promptly are essential for an optimal clinical outcome. PMID- 19306158 TI - Physical activity and quality of life in subjects with chronic disease: chronic obstructive pulmonary disease compared with rheumatoid arthritis and diabetes mellitus. AB - OBJECTIVE: Chronic diseases interfere with the life situation of the affected person in different ways. The aim was to compare the burden of disease in three chronic diseases - chronic obstructive pulmonary disease (COPD), rheumatoid arthritis (RA), diabetes mellitus (DM) - and in healthy subjects, with a particular interest in physical activity, quality of life, and psychological health. DESIGN: Cross-sectional, observational study. SETTING AND SUBJECTS: Postal survey questionnaire to a stratified, random population of 68 460 subjects aged 18-84 years in Sweden. The subjects included were 40-84 years old (n = 43 589) and data were analysed for COPD (n = 526), RA (n = 1120), DM (n = 2149) and healthy subjects (n = 6960). RESULT: Some 84% of subjects with COPD, 74% (RA), 72% (DM), and 60% in healthy subjects (p < 0.001, COPD versus RA, DM, and healthy subjects) had a physical activity level considered too low to maintain good health according to guidelines. Quality of life (EuroQol five-dimension questionnaire, EQ-5D) was lower in COPD and RA than in DM. Anxiety/depression was more common in subjects with COPD (53%) than in those with RA (48%) and DM (35%) (p < 0.001, COPD versus RA and DM), whereas mobility problems were more common in RA (55%) than COPD (48%) and DM (36%) (p < 0.001, RA versus COPD and DM). All differences between groups remained significant after adjusting for age, sex, and socioeconomic background factors. CONCLUSION: Subjects with chronic diseases had a low level of physical activity, most evident in subjects with COPD. COPD and RA had a higher negative impact on quality of life than DM. Our results indicate that increased attention regarding physical inactivity in subjects with chronic diseases is needed to minimize the burden of disease. PMID- 19306159 TI - Association of hypoxia-inducible factor 1A (HIF1A) gene polymorphisms with systemic sclerosis in a French European Caucasian population. AB - OBJECTIVE: Systemic sclerosis (SSc) is a connective tissue disease characterized by generalized microangiopathy leading to chronic hypoxia. The aim of this study was to determine whether polymorphisms of the hypoxia-inducible factor 1A gene (HIF1A) affects susceptibility to SSc in a large French European Caucasian population. METHODS: A case-control study was performed in 659 SSc patients and 511 healthy matched controls. Three tag single nucleotide polymorphisms (SNPs) of the HIF1A gene (rs12434438 A/G, rs1957757 C/T, and rs11549465 C/T) were genotyped allowing whole gene coverage according to HapMap data. RESULTS: The frequency of genotypes carrying at least one G allele (A/G and/or GG) of the rs12434438 SNP was significantly higher in SSc patients than in controls [p(corr) = 0.018, odds ratio (OR) 1.44, 95% confidence interval (CI) 1.08-1.91]. Regarding SSc subgroup analyses, the heterozygous genotype A/G was associated with SSc (p(corr) = 0.012, OR 1.47, 95% CI 1.13-1.9), with the limited cutaneous form of SSc (p(corr) = 0.04, OR 1.43, 95% CI 1.08-1.91), and with positive anti-centromere antibodies (ACA; p(corr) = 0.016, OR 1.61, 95% CI 1.16-2.23). No association was detected for the remaining two HIF1A SNPs tested. Haplotype analyses did not detect any association with SSc. CONCLUSIONS: We observed an association between the HIF1A gene and SSc in a European Caucasian population, supporting a role for HIF1 in the pathophysiology of SSc. PMID- 19306160 TI - High-resolution melting analysis for the rapid detection of an intronic single nucleotide polymorphism in SLC22A12 in male patients with primary gout in China. AB - OBJECTIVES: The human urate transporter 1 (URAT1, encoded by SLC22A12) was recently identified as the major absorptive urate transporter protein in the kidney responsible for regulating blood urate levels. The present study was designed to investigate the rs893006 polymorphism (GG, GT, and TT) in SLC22A12 in a total of 292 Chinese male subjects. Differences of clinical characteristics among the genotype groups were analysed. METHODS: A total of 124 consecutive patients with diagnosis of primary gout and 168 healthy male volunteers were enrolled in this study. Demographic and clinical data were obtained from the patients and controls. DNA was purified from peripheral blood and the rs893006 polymorphism was determined with sequencing analysis. In addition, DNA samples were detected by high-resolution melting (HRM) analysis. Melting curves were analysed as fluorescence difference plots. The shift and curve shapes of melting profiles were used to distinguish the different genotypes. RESULTS: GG, GT, and TT genotypes were unambiguously distinguished with HRM technology. Genotyping based on HRM analysis was fully concordant with the sequencing. Serum uric acid levels in the TT genotype subjects were significantly lower than those in the GG and GT genotypes. However, no differences among the groups were found in body mass index (BMI), blood pressure, creatinine, total cholesterol, and triglycerides. The TT genotype was observed more frequently among the low uric acid group than the high uric acid group. CONCLUSIONS: HRM analysis is a simple, rapid and accurate one-tube assay for genotyping the SLCSSA12 gene. The rs893006 polymorphism in SLC22CA12 was confirmed to be a genetic risk for hyperuricaemia among the Chinese male population. PMID- 19306161 TI - Decision by sampling: the role of the decision environment in risky choice. AB - Decision by sampling (DbS) is a theory about how our environment shapes the decisions that we make. Here, I review the application of DbS to risky decision making. According to classical theories of risky decision making, people make stable transformations between outcomes and probabilities and their subjective counterparts using fixed psychoeconomic functions. DbS offers a quite different account. In DbS, the subjective value of an outcome or probability is derived from a series of binary, ordinal comparisons with a sample of other outcomes or probabilities from the decision environment. In this way, the distribution of attribute values in the environment determines the subjective valuations of outcomes and probabilities. I show how DbS interacts with the real-world distributions of gains, losses, and probabilities to produce the classical psychoeconomic functions. I extend DbS to account for preferences in benchmark data sets. Finally, in a challenge to the classical notion of stable subjective valuations, I review evidence that manipulating the distribution of attribute values in the environment changes our subjective valuations just as DbS predicts. PMID- 19306162 TI - Relationships among spiritual beliefs, religious practises, congregational support and health for individuals with traumatic brain injury. AB - OBJECTIVE: To determine relationships among spiritual beliefs, religious practises, congregational support and health for individuals with traumatic brain injury (TBI). DESIGN: A cross-sectional analysis of 61 individuals with TBI evaluated in an outpatient clinic using the Brief Multidimensional Measure of Religiousness/Spirituality (BMMRS) and the Medical Outcomes Scale-Short Form 36 (SF-36). RESULTS: For persons with TBI the BMMRS Meaning and Values/Beliefs sub scales were significantly correlated with the SF-36 General Health Perception sub scale and the BMMRS Religious Support sub-scale was significantly correlated with the SF-36 General Mental Health sub-scale. Hierarchical regressions indicated that the BMMRS Values/Beliefs and Forgiveness sub-scales accounted for 16% additional variance in SF-36 General Health Perception scores beyond that accounted for by demographic variables (i.e. age, income); no BMMRS sub-scales accounted for additional variance in predicting the SF-36 General Mental Health sub-scale beyond that accounted for by demographic variables (i.e. age, income). CONCLUSIONS: The physical health of individuals with TBI is associated with spiritual beliefs but not religious practises or congregational support. Better mental health is associated with increasing congregationally based social support for persons with TBI. Religious practises (i.e. praying, etc.) are not related to either physical or mental health, as some persons with TBI may increase prayer with declining health status. PMID- 19306163 TI - External influences on the safe practice of neurosurgery. PMID- 19306164 TI - Patient safety with particular reference to wrong site surgery--a presidential commentary. PMID- 19306168 TI - Biographical sketch: F. John Gillingham. PMID- 19306169 TI - The current status of decompressive craniectomy. AB - Decompressive Craniectomy (DC) continues to be widely practiced but remains controversial. The procedure has its origins thousands of years ago, with early trepanation performed for a range of medical and religious reasons. We summarize the history, techniques, complications and pathophysiology and then explore in detail the recent evidence base for the most common indications for DC; Traumatic brain injury (TBI) and Cerebral infarction. An important consideration from the outset is the often forgotten issue of cranioplasty and we summarize advances in materials, technology and discuss the optimum timing. Outcomes of ongoing randomized trials in TBI are awaited with interest but the trend in the nonrandomized literature suggests timely intervention reduces mortality with acceptable morbidity. Level 1 evidence for early DC in young patients with malignant middle cerebral artery infarction has arrived and has implications for neurosurgical practice and rehabilitation services. Current European and North American practice recommends the judicious use of DC in traumatic brain injury and malignant middle cerebral artery infarction in select patients. PMID- 19306170 TI - Emergency head CT scans: can neurosurgical registrars be relied upon to interpret them? AB - Neurosurgical registrars are frequently called upon by A&E staff and physicians to interpret emergency head CT (computed tomography) scans out of hours. This appears to reflect the reduced threshold for scanning patients and the nonavailability of a radiologist to report these scans. This study was undertaken to assess the safety of such practices. Five neurosurgical registrars, blinded to each other and to the radiology reports, interpreted 50 consecutive emergency head CT scans (both trauma and nontrauma) from the hospital's imaging system as a pilot study. These were initially graded as normal or abnormal. Abnormal scans were assessed for the presence of an intracranial bleed, pneumocephalus, skull fractures, cerebral contusions, mass effect, midline shift, ischaemia or hydrocephalus. The agreement of the observers' recordings with the report issued or approved by a consultant radiologist was evaluated using SPSS Version 13.0. Four of the five registrars assessed a further 150 scans in a similar manner to complete the study. There was a good general agreement between the formal reports and the neurosurgical registrars' identification of normal scans (average Kappa 0.79). The radiology reports and the registrars also agreed well on the presence or absence of intracranial blood, contusions and pneumocephalus (Kappa value > 0.70). The agreement was poorer for ischaemia, mass lesions (other than intracranial haematomas), grey white differentiation, evidence of raised intracranial pressure and midline shift (Kappa < 0.5). Neurosurgical registrars compared well with radiologists when it came to assessing emergency head CT scans as normal or detecting a surgical lesion. The agreement was poorer on subtle abnormalities. The practice of neurosurgical registrars informally 'reporting' on emergency head CT scans cannot be recommended. PMID- 19306171 TI - The burden of motorcycle-related neuro-trauma in Ireland and associated helmet usage. AB - Motorcycles represent less than 2% of the licensed vehicles but motorcyclists account for 12% of road deaths in Ireland. The British Road Safety Authority has introduced the Sharp programme, which hopes to save 50 lives in the U.K. each year alone by helping riders to choose the best-fitting and safest helmets. We evaluated the pattern of head injuries sustained by motorcyclists referred to the two neurosurgical centres Beaumont Hospital and Cork University Hospital in Ireland and ascertained if the new SHARP guidelines could be of benefit in reducing the burden of motorcycle related neurotrauma and disability in Ireland. Despite Ireland having mandatory helmet laws almost a quarter of our motorcyclists with traumatic brain injury were unhelmeted. A significant reduction in mortality and morbidity is predicted if all motorcyclists in Ireland were to wear helmets that satisfied the SHARP criteria. PMID- 19306172 TI - Gunshot injuries to the brachial plexus during wartime. AB - Gunshot injuries of the brachial plexus are a challenging issue among peripheral nerve lesions. Surgical reconstruction of such injuries is difficult and the clinical outcome depends on several factors. The aim of this study was to present the outcome of surgical management of gunshot injuries of the brachial plexus that occurred following Iran-Iraq war. Twenty patients with 55 injured elements of the brachial plexus underwent surgery in Loghman-Hakim Hospital during 1982 and 1992. Reconstructive procedures included neurolysis in 30 injured elements, nerve grafting in 17 and a combination of these two methods in 8 cases. Surgical procedure was selected based on the microscopic findings during the operation. Final recovery outcome was assessed at least 3 years after surgery on the basis of motor and sensory recoveries. Final outcome was defined as poor, intermediate, and good. Both good and intermediate outcomes were considered as useful recovery. An acceptable recovery was obtained in 28 of 30 (94%) injured elements undergone neurolysis, 15 of 17 (89%) elements in nerve graft group, and 7 of 8 (87.5%) elements reconstructed with neurolysis in combination with nerve graft. In neurolysis, good recovery was more frequent and obtained in 23 of 30 (77.5%) lesions. Best treatment outcome was observed in lesions of lateral cord to musculocutaneous nerve which all injured elements showed good recovery. Impairment in none of the lesions in the level of posterior cord and lower trunk or C8-T1 led to good recovery. In surgical reconstruction of gunshot injuries of the brachial plexus the most favorable results were observed in the neurolysis reconstruction of the lesions in the lateral cord to musculocutaneous nerve. In the absence of spontaneous improvement of neurologic deficit, surgical procedures should be done as soon as possible according to the type and location of injury. PMID- 19306173 TI - Relationship between volume, dose and local control in stereotactic radiosurgery of brain metastasis. AB - The aim of this study is to analyse the efficacy of linear accelerator stereotactic radiosurgery (SRS) on prognostic factors, local control rate and survival in patients with brain metastasis. Patients with either a single metastasis or up to 4 multiple brain metastases with a maximum tumour diameter of 40 mm for each tumour and a Karnofsky Performance Status (KPS) > or = 70 were eligible for SRS. SRS was applied to 150 lesions in 86 consecutive patients with a median age of 60 years (median 1 and mean 1.7 lesions per patient, mean KPS 86). Median overall survival was 6.2 months after SRS and 9.7 months from diagnosis of brain metastasis. Multivariate analysis revealed that a KPS of 90 or more (p = 0.009) and female sex (p = 0.003) were associated with a longer survival. Radiation dose < or = 15 Gy (p = 0.017) and KPS < 90 (p = 0.013) were independent predictors of a shorter time to local failure. Five patients showed evidence of radionecrosis with a median survival of 14.8 months. Addition of WBRT neither led to improvement of survival nor to improvement of local control. Improved local control following SRS for brain metastases was associated with KPS > or =90, a radiation dose > 15 Gy and a PTV < 13 cc. The potential of hypofractionated stereotactic radiotherapy (SRT) for brain metastases of larger volume warrants further study. PMID- 19306174 TI - The incidence of seizures following Deep Brain Stimulating electrode implantation for movement disorders, pain and psychiatric conditions. AB - Deep Brain Stimulation (DBS) for neuromodulation is now commonplace. However little is known about the incidence of either procedural related seizures or epilepsy following chronic DBS. This study aims to provide estimates of these complications for movement disorders, pain and psychiatric conditions. A literature review was performed. Because searches using the terms seizure, epilepsy, and deep brain stimulation revealed only papers dealing with experimental and clinical application of DBS to treat chronic seizures disorders, a search strategy trawling through papers that described clinical case series of DBS was used. Thirty-two papers were reviewed that described stereotactic placement of DBS electrodes for movement disorders, pain syndromes and psychiatric conditions with cohorts of n > 5. Sixteen of these papers describing at least 1418 DBS electrode placements in 1254 patients did not mention seizures as a complication (i.e., it was not possible to know whether seizures had or had not occurred). In 16 papers, describing at least 2101 electrode placements in 1555 patients, seizures were described in 42 patients (incidence 2.7%). The range of seizure incidence varied from 0% (three series encompassing 317 patients and 576 electrode placements) up to 10% (n = 130) and 13% (n = 15). The reasons for this variance were not obvious. At least 74% of seizures occurred around the time of electrode implantation and many of these patients also suffered intracranial hemorrhage. Follow up times were variable (range 6 mths to 5 years). The analysis was complicated by multiple publications from some centres with duplication of some data. The quality of literature on seizures following DBS insertion for neuromodulation is highly variable. Analysis of the available data, after making corrections for publication of duplicate data, suggests strongly that the risk of seizures associated with DBS placement is probably lower than 2.4% (95% CI 1.7 to 3.3 %). The risk of postprocedural seizures associated with chronic deep brain stimulation is even lower with best estimates around 0.5% (95% CI .02 to 1.0%). PMID- 19306175 TI - Fitness to fly post craniotomy--a survey of medical advice from long-haul airline carriers. AB - Commercial airline passengers are subject to numerous medical risks while in transit. Seventeen long-haul airline companies were questioned concerning fitness to travel and the case of a patient wishing to travel post craniotomy. Three airline companies gave satisfactory medical information, while the remaining airlines felt it was the decision of the operating surgeon rather than the airline company. A literature review shows that post operative pneumocephalus and the risk of tension pneumocephalus is the major medical concern when transporting patients post craniotomy. Evidence is contradictory with respect to the importance of this potentially life threatening problem. Postoperative 100% oxygen may improve the rate of pneumocephalus absorption. Airline companies have an unstandardised approach to unique medical problems, resulting in increased responsibility for the attending surgeon who may be ill equipped to deal with poorly researched aviation medicine. PMID- 19306176 TI - The Fisher grading correlated to outcome in patients with subarachnoid haemorrhage. AB - Cerebral vasospasm is the major cause of delayed ischemia in patients with subarachnoid haemorrhage (SAH). The Fisher grading scale has been used to predict patients in risk of developing vasospasm. Improved radiological techniques and treatment may have changed the relevance of the Fisher scale. We have now evaluated the Fisher scale, Hunt and Hess and age in relation to outcome in patients with SAH. Eighty- three patients were admitted with SAH during two years, and 84 aneurysms were treated in 78 patients. The Glasgow outcome score (GOS) within 3 months were as follows; GOS 1 (19%), GOS 2 (2%), GOS 3 (11%), GOS 4 (9%), GOS 5 (59%). There was a significant correlation between both the Fisher grading scale, Hunt and Hess scale and outcome. Age was not correlated to the Fisher grading scale or the Hunt and Hess scale. Age was also not correlated to outcome in our patients. Despite the correlation to outcome both Hunt and Hess and the Fisher grading scale had a limited predictive value of outcome due to a low specificity and/or sensitivity. PMID- 19306177 TI - Neurosciences intensive care medicine in initial neurosurgical training. AB - The authors describe a novel 4-month clinical placement in neurosciences intensive care medicine (NICM) undertaken in the first specialty registrar (ST1) year of neurosurgical training as part of a clinical neurosciences themed training year. Neurosurgery is unique among British surgical specialties in having pioneered themed early years in run-through training to replace basic surgical training in general surgical specialties as part of Modernising Medical Careers. After describing events leading to the new neurosurgical training, the knowledge, skills and attitudes acquired in NICM are highlighted alongside discussion of logistic aspects and future directions from an inaugural experience. PMID- 19306178 TI - Management of associated glioma and arteriovenous malformation--the priority is the glioma. AB - The conjunction of a glioma with an arteriovenous malformation (AVM) is exceptionally rare. We report the case of a malignant pleomorphic xanthoastrocytoma located on the vicinity of an untreated AVM that was removed without interference with the AVM. The question posed by such concurrent lesions is which to manage first. It appears more logical to treat the tumour first because of the elevated incidence of associated high-grade gliomas. PMID- 19306179 TI - Carcinoid tumour in a lumbar teratoma associated with tethered cord syndrome in an adult. AB - An unusual case of a carcinoid tumour in a lumbar extradural teratoma presenting as tethered cord syndrome and spina bifida without carcinoid syndrome in a 51 year-old woman is reported. These entities existing simultaneously in the same region are extremely rare and this suggests they have a common origin. PMID- 19306180 TI - Cerebellar tuberculoma presenting as haematoma--a case report and pathophysiological consideration. AB - Intracranial tuberculoma represents the most common mass lesion in neurotuberculosis. We report an unusual case of tuberculoma with haemorrhage presenting clinically as a cerebellar haemorrhage. Cranial CT scan revealed a peripherally enhancing cerebellar mass with haemorrhage and obstructive hydrocephalus, requiring urgent surgical evacuation of the haematoma. Histopathological examination revealed tuberculoma with features of phlebitis. In view of the pathological evidence of phlebitis in the lesion, it could represent an immune-mediated acute haemorrhagic leucoencephalitis in response to tubercular antigens. PMID- 19306181 TI - Rapid growth of small, asymptomatic meningioma following radiosurgery. AB - The authors present the case of a 62-year-old woman with rapid enlargement of a meningioma following radiosurgery (RS). Previous slow growth of the tumor over a 3-year period and the radiological signs of benign meningioma had been confirmed by successive MR scans. Histopathological examination performed after successful surgical removal revealed an atypical, infiltrating meningioma. PMID- 19306182 TI - Pitfall of CT angiography for an aneurysm around middle cerebral artery bifurcation. AB - Computerized tomography(CT) angiography is a reliable alternative to digital subtraction angiography for the detection of aneurysms. The authors report an aneurysm of anterior temporal artery(ATA) misinterpreted as being in the bifurcation of middle cerebral artery(MCA) treated by aneurysm excision followed by primary anastomosis. PMID- 19306183 TI - Pin-site epidural hematoma in an adult case of chronic hydrocephalus with associated thinning of the cranium. AB - We report a case of a 22-year-old man presenting with a fourth ventricular tumour and associated chronic obstructive hydrocephalus likely leading to thinning of the cranium. Intraoperatively, he developed an epidural hematoma secondary to a fracture of the temporal bone by application of the three-point skull fixator. This is the second reported adult case of an iatrogenic epidural hematoma secondary to pin-site complications. We advocate the careful placement of the pins and suggest the tightening force be catered individually especially in pediatric patients or patients with chronic hydrocephalus. PMID- 19306184 TI - Contralateral acute interdural haematoma occurring after burr hole drainage of chronic subdural haematoma. AB - We report the case of a 78-year-old man with chronic subdural haematoma (CSDH) who presented with impairment in recent memory and gait disturbance. He underwent burr-hole craniostomy with a closed-drainage system. A computed tomography scan conducted on postoperative day 3 demonstrated an acute epidural haematoma over the contralateral frontoparietal convexity. Craniotomy and haematoma evacuation were immediately performed. The haematoma was located between the outer and inner dura mater that each comprise a single layer. To our knowledge, this is the first reported case of an acute haematoma located between the separated dura mater that occurred following drainage of a contralateral CSDH, and it is the second reported case of interdural haematoma over the cerebral convexity. PMID- 19306185 TI - Picture quiz: April 2009--trauma. AB - This month's picture quiz provides a series of questions to test practising neurosurgeons and neurosurgeons in training on trauma. PMID- 19306188 TI - SBNS survey of CSBH management. PMID- 19306190 TI - Electrophysiological correlates of intermediate syndrome following acute organophosphate poisoning. AB - INTRODUCTION: Organophosphate (OP) poisoning is a major global health problem. The late onset of respiratory failure associated with intermediate syndrome (IMS) is a major contributor to the high morbidity, mortality, and cost of OP poisoning. This is particularly important as most poisoning occurs in the under resourced developing world. Repetitive nerve stimulation studies. An understanding of the abnormalities observed in repetitive nerve stimulation studies during the progression and development of IMS spectrum disorder may help clinicians to utilize electrodiagnostic testing in the better management of their patients with acute OP poisoning. In addition, it will allow researchers to interpret future research that utilizes repetitive nerve stimulation as an outcome measure. A review of the clinical and experimental electrophysiological studies in the IMS shows that subclinical electrophysiological abnormalities are common, progressive, and precede the onset of the clinical IMS. Serial repetitive nerve stimulation studies have been most commonly used and are the most accessible for clinicians. Clinical and experimental studies demonstrate a progression through early initial decrement-increment patterns at high rates of stimulations, which correlate with moderate muscle weakness, to decrement increment patterns at intermediate- and low-frequency stimulations. Progression to a combination of decrement-increment and repetitive fade patterns correlates with clinical deterioration; severe decrement pattern is usually observed immediately before the onset of respiratory failure. Although electrophysiological features closely parallel clinical severity during progression of IMS, the same is not true during recovery. Electrophysiological changes sometimes improve long before the patient recovers normal strength and respiratory function. Intermediate syndrome. Thus, IMS can be regarded as a spectrum disorder affecting the neuromuscular junction (NMJ) with two main forms: a forme fruste variety associated with mild weakness and the classical IMS with weakness of 3/5 or less than 3/5 on the Medical Research Council (MRC) grading; patients in the latter category are at risk of developing late onset respiratory failure. While IMS remains a clinically important entity, the early occurrence of abnormalities on repetitive nerve stimulation studies suggest that this is part of the continuum of nicotinic receptor stimulation. CONCLUSIONS: Reviewing the anatomical and the functional structure of the NMJ and neuromuscular transmission helps to provide an understanding of the pathophysiological nature of the neuromuscular transmission failure observed in IMS. This includes potential mechanisms of presynaptic feedback which may reduce acetylcholine release and postsynaptic receptor desensitization and provides some explanation for the time course of IMS. It also suggests other potential strategies to reduce OP-induced NMJ toxicity in which repetitive nerve stimulation is likely to be an important tool in judging efficacy. PMID- 19306191 TI - Management of yellow oleander poisoning. AB - BACKGROUND: Poisoning due to deliberate self-harm with the seeds of yellow oleander (Thevetia peruviana) results in significant morbidity and mortality each year in South Asia. Yellow oleander seeds contain highly toxic cardiac glycosides including thevetins A and B and neriifolin. A wide variety of bradyarrhythmias and tachyarrhythmias occur following ingestion. Important epidemiological and clinical differences exist between poisoning due to yellow oleander and digoxin; yellow oleander poisoning is commonly seen in younger patients without preexisting illness or comorbidity. Assessment and initial management. Initial assessment and management is similar to other poisonings. No definite criteria are available for risk stratification. Continuous ECG monitoring for at least 24 h is necessary to detect arrhythmias; longer monitoring is appropriate in patients with severe poisoning. Supportive care. Correction of dehydration with normal saline is necessary, and antiemetics are used to control severe vomiting. Electrolytes. Hypokalemia worsens toxicity due to digitalis glycosides, and hyperkalemia is life-threatening. Both must be corrected. Hyperkalemia is due to extracellular shift of potassium rather than an increase in total body potassium and is best treated with insulin-dextrose infusion. Intravenous calcium increases the risk of cardiac arrhythmias and is not recommended in treating hyperkalemia. Oral or rectal administration of sodium polystyrene sulfonate resin may result in hypokalemia when used together with digoxin-specific antibody fragments. Unlike digoxin toxicity, serum magnesium concentrations are less likely to be affected in yellow oleander poisoning. The effect of magnesium concentrations on toxicity and outcome is not known. Hypomagnesaemia should be corrected as it can worsen cardiac glycoside toxicity. Gastric decontamination. The place of emesis induction and gastric lavage has not been investigated, although they are used in practice. Gastric decontamination by the use of single dose and multiple doses of activated charcoal has been evaluated in two randomized controlled trials, with contradictory results. Methodological differences (severity of poisoning in recruited patients, duration of treatment, compliance) between the two trials, together with differences in mortality rates in control groups, have led to much controversy. No firm recommendation for or against the use of multiple doses of activated charcoal can be made at present, and further studies are needed. Single dose activated charcoal is probably beneficial. Activated charcoal is clearly safe. Arrhythmia management. Bradyarrhythmias are commonly managed with atropine, isoprenaline, and temporary cardiac pacing in severe cases, although without trial evidence of survival benefit, or adequate evaluation of possible risks. Accelerating the heart rate with atropine or beta-adrenergic agents theoretically increases the risk of tachyarrhythmias, and it has been claimed that atropine increases tachyarrhythmic deaths. Further studies are required. Tachyarrhythmias have a poor prognosis and are more difficult to treat. Lidocaine is the preferred antiarrhythmic; the role of intravenous magnesium is uncertain. Digoxin-specific antibody fragments. Digoxin-specific antibody fragments are effective in reverting life-threatening cardiac arrhythmias; prospective observational studies show a beneficial effect on mortality. High cost and lack of availability limit the widespread use of digoxin-specific antibody fragments in developing countries. CONCLUSIONS: Digoxin-specific antibody fragments remain the only proven therapy for yellow oleander poisoning. Further studies are needed to determine the place of activated charcoal, the benefits or risks of atropine and isoprenaline, the place and choice of antiarrhythmics, and the effect of intravenous magnesium in yellow oleander poisoning. PMID- 19306192 TI - Regional variation in the severity of pesticide exposure outcomes: applications of geographic information systems and spatial scan statistics. AB - INTRODUCTION: Geographic information systems and spatial scan statistics have been utilized to assess regional clustering of symptomatic pesticide exposures reported to a state Poison Control Center (PCC) during a single year. In the present study, we analyzed five subsequent years of PCC data to test whether there are significant geographic differences in pesticide exposure incidents resulting in serious (moderate, major, and fatal) medical outcomes. METHODS: A PCC provided the data on unintentional pesticide exposures for the time period 2001-2005. The geographic location of the caller, the location where the exposure occurred, the exposure route, and the medical outcome were abstracted. RESULTS: There were 273 incidents resulting in moderate effects (n = 261), major effects (n = 10), or fatalities (n = 2). Spatial scan statistics identified a geographic area consisting of two adjacent counties (one urban, one rural), where statistically significant clustering of serious outcomes was observed. The relative risk of moderate, major, and fatal outcomes was 2.0 in this spatial cluster (p = 0.0005). CONCLUSIONS: PCC data, geographic information systems, and spatial scan statistics can identify clustering of serious outcomes from human exposure to pesticides. These analyses may be useful for public health officials to target preventive interventions. Further investigation is warranted to understand better the potential explanations for geographical clustering, and to assess whether preventive interventions have an impact on reducing pesticide exposure incidents resulting in serious medical outcomes. PMID- 19306194 TI - Cardiovascular diagnostics mining on Borneo island: from labour-intensive to prototype. AB - Mining for medical data poses different challenges compared with mining other types of data. The wide range of imaging modalities of medical data leads to data integration and compatibility issues. The analysis of imaging modalities is further complicated by the different format and attributes used by the different imaging equipment by different vendors. Human factors such as interest of adapting data mining into diagnosis and planning process raised the difficulty of engaging the users into the development of a practical and useful data miner. Requirement engineering technique prototyping further enhanced the engagement of users towards the data-miner. Data from different equipment and different vendors are also merged for efficient data analysis and subsequently charting and reporting. We have also successfully engaged the medical doctors into believing the data miner's capability after they reviewed and walkthrough the prototype. PMID- 19306195 TI - Quality of cystic fibrosis information on Italian websites. AB - Given the increased attention on cystic fibrosis care during the last few years in Italy, this study was performed to evaluate content quality, characteristics and completeness of Italian cystic fibrosis websites. GOOGLE was used and retrieved websites were evaluated for the presence of predefined descriptive criteria and disease information. Their contents were compared with available evidence. Thirty sites were evaluated. Few fulfilled the criteria, such as listing authors (24 sites) and citing references (10). The amount of disease information varied, but 23 provided treatment information. Of these, 11 comparable sites were evaluated for adherence to guidelines and overall completeness. All information adhered to guidelines. One site provided very thorough information and was the most complete, whereas four were found to be half as complete as others. Patients can find accurate cystic fibrosis knowledge online, but may need to navigate between multiple sites to obtain comprehensive coverage of disease information. In Italy, a network of association websites exists to support patients and families. Website quality could be improved in terms of documentation (transparency) of the information provided in order to permit users to select well-documented, updated information. PMID- 19306196 TI - Health-related Internet use by patients with somatic diseases: frequency of use and characteristics of users. AB - The aim of this study was to explore the percentage of Dutch patients using the Internet to search for information about their illness. In addition, we studied patients' usage of health-related Internet applications, such as online patient support groups. The final objective of this study was to explore which demographic, health and psychological characteristics are related to patients' health-related Internet use. In order to answer these research questions we sent a written questionnaire to a representative sample of patients with breast cancer, rheumatoid arthritis and fibromyalgia. The overall total response rate was 69% (N = 679). In total, 52% of the patients used the Internet to search for information about their illness. However, only 15% of the respondents had ever read along with an online patient support group. An even smaller proportion of the patients (4%) had send postings to such a group. Online communication with health professionals was not commonly practiced. A younger age, a higher education and employment appeared to be the only significant predictors of patients' health-related Internet use. Patients' health-related Internet use could not be predicted by health and psychological characteristics. Although, about half of the patients made use of the Internet for health-related reasons, mostly their health-related Internet use was restricted to seeking information about their illness. PMID- 19306197 TI - User information seeking behaviour: perceptions and reality. An evaluation of the WHO Labresources Internet portal. AB - Evaluation on Internet portals is a key component of any online resource development. Understanding user information seeking behaviour and user perceived behaviour is essential to obtain the full picture of user needs, online activities and draw lessons to improve the design of Internet portals to better meet user expectations. This article discusses the evaluation of a WHO Internet portal: the Labresources website. The evaluation investigates user satisfaction with the resource, usability, demographic information about users and how well they could complete specific tasks using the website and compared this with the actual online behaviour revealing a number of discrepancies. An online questionnaire was advertised on the Labresources website during the period 25 November 2005 to 20 February 2006. As the site caters to English and French speakers, the questionnaire was made available in both languages. It consisted of two sections - the first section required the participant to complete three tasks using the website whereas the second section tested user satisfaction, information needs and appropriateness of the content. Weblogs data were compared with the questionnaire results to compare user perceived and actual online behaviour. Twenty one respondents completed the online questionnaire from a total of 18 countries. This was out of a potential 60 website users among whom the questionnaire was promoted. In general, respondents were satisfied with the website layout and navigation. 61.9% of respondents listed WHO among their top 5 and a third listed the Labresources website. The number of sessions where users browse (146) the information resources is almost three times more than the number of users who search (52) the resources. Weblogs revealed most interesting results with differences between what users reported doing when completing tasks and how easy they perceived the tasks and what they actually did. Twelve respondents completed at least one task. Of the remaining nine respondents, three did visit the Labresources website from the link in the questionnaire but did not go on to complete the tasks. Only one of the twelve who completed a task reported it being difficult. Three of the respondents who didn't complete a task reported the tasks to be difficult but only one of these actually followed the link to begin the tasks in the questionnaire. This article described an evaluation study investigating user perceived and actual behaviour at the WHO Labresources Internet portal. Although the questionnaire results demonstrated general satisfaction with the resource, a combination of a weblog evaluation with the questionnaire revealed a clearer picture of the user perception of and satisfaction with the website compared to their actual activity when completing the set tasks. PMID- 19306198 TI - Influences on physicians' adoption of electronic detailing (e-detailing). AB - E-detailing means using digital technology: internet, video conferencing and interactive voice response. There are two types of e-detailing: interactive (virtual) and video. Currently, little is known about what factors influence physicians' adoption of e-detailing. The objectives of this study were to test a model of physicians' adoption of e-detailing and to describe physicians using e detailing. A mail survey was sent to a random sample of 2000 physicians practicing in Iowa. Binomial logistic regression was used to test the model of influences on physician adoption of e-detailing. On the basis of Rogers' model of adoption, the independent variables included relative advantage, compatibility, complexity, peer influence, attitudes, years in practice, presence of restrictive access to traditional detailing, type of specialty, academic affiliation, type of practice setting and control variables. A total of 671 responses were received giving a response rate of 34.7%. A total of 141 physicians (21.0%) reported using of e-detailing. The overall adoption model for using either type of e-detailing was found to be significant. Relative advantage, peer influence, attitudes, type of specialty, presence of restrictive access and years of practice had significant influences on physician adoption of e-detailing. The model of adoption of innovation is useful to explain physicians' adoption of e-detailing. PMID- 19306199 TI - Reliability of short form-36 in an Internet- and a pen-and-paper version. AB - Use of Internet versions of questionnaires may have several advantages in clinical and epidemiological research, but we know little about if Internet versions differ with respect to validity and reliability. We aimed to compare Internet- and pen-and-paper versions of short form-36 (SF-36) with respect to test-retest reliability and internal consistency. Women referred to mammography (n = 782) were randomised to receive either a paper version with a prepaid return envelope or a guideline on how to fill in the Internet version. A subgroup was asked to answer the questionnaire once again in the alternative version. Test retest reliability was assessed by the intra-class correlation coefficient. Internal consistency was calculated as Cronbach's alpha. The between-version test retest reliability for the eight subscales were between 0.63 and 0.92. Cronbach's alpha for the two versions were all between 0.75 and 0.93 with minor differences between the Internet- and the pen-and-paper version. We found little or no evidence of a difference in test-retest reliability and internal consistency when we compared an Internet- and a pen-and-paper version of SF-36. PMID- 19306200 TI - Functional health literacy and cancer care conversations in online forums for retired persons. AB - Cancer is primarily a chronic disease of older adults that must be managed and incorporated into everyday activities. Online sites are important sources of health care information and support. Health literacy is necessary for full utilisation of online resources. The objective of this study was to examine and compare cancer related conversations in online forums hosted by Canadian and American associations for retired persons. A content analysis was used to evaluate archived cancer conversations of general-health online forums representing two leading North American associations for retired persons. There were 125/1817 (6.8%) Canadians and 70/892 (7.8%) US cancer discussions among participants in 2006. Online conversations were grouped into three categories: request for information, provision of information and sharing of information. Important subthemes included cancer prevention and screening, treatment and cancer care and health system issues. There were significantly more posts about provision of cancer information from the Canadian compared with the US site (p = 0.023). American more than Canadian conversations emphasised the health system concerns as determinants of cancer care practices. Online discussion forums hosted by retirement associations may serve as an important channel in information dissemination about cancer prevention and screening, treatment and care support and health care advice for seniors. PMID- 19306201 TI - Implementation of an onto-wiki toolkit using web services to improve the efficiency and effectiveness of medical ontology co-authoring and analysis. AB - The ICD11 draft was launched by the WHO in order to define ICD ontology by selected experts using a wiki-like structured joint-authoring tool. The challenge of this expert-/community-based ontology co-authoring is how to manage and process the ontology objects in the wiki page. The wiki-tools in the market require lots of time and human effort to organise, process and extract the ontology content from the wiki page for review and analysis. Therefore, this article is to investigate how to apply semantic web technology to improve the efficiency and effectiveness of ontology co-authoring and analysis. An onto-wiki toolkit that provides a set of web services is proposed for ontology creators to create, co-edit, organise, map and relate the wiki-like structured ontology objects on a wiki page for review and analysis. By using the onto-wiki toolkits, the pace of terminology standardisation, e-patient records integration, exchange and sharing can be improved. PMID- 19306202 TI - Recidivism among child sexual abusers: initial results of a 13-year longitudinal random sample. AB - In the initial analysis of data from a random sample of all those charged with child sexual abuse in Idaho over a 13-year period, only one predictive variable was found that related to recidivism of those convicted. Variables such as ethnicity, relationship, gender, and age differences did not show a significant or even large association with recidivism. The only variable that seemed to show both a significant and almost moderate association to recidivism was the Risk Assessment in the Sex Offender Evaluation reoffense. Comparisons were made to prior research as well as a discussion of implications of the sex offender evaluation for the legal process. Finally, a call for the continued need for further research is discussed. PMID- 19306203 TI - A comparative study of two groups of sex offenders identified as high and low risk on the static-99. AB - The purpose of this study was to identify possible differences between high- and low-risk sex offenders. The subjects included 285 sex offenders on probation. They were evaluated with the Static-99, Abel Assessment, Raven's, and MMPI-2. A criminal history review identified the number of prior offenses and the age/sex category in the index offense. The high- and low-risk groups were compared on 26 variables: intelligence, age, criminal history, denial patterns, measured sexual interest in children, admission of sexual interests, a childhood history of sexual abuse, victim's age, and personality variables. Four variables significantly accounted for 64% of the variance: age, prior number of felonies, the Cognitive Distortion Score, and the MMPI-2 Infrequency scale score. PMID- 19306204 TI - Male juveniles with sexual behavior problems: are there differences among racial groups? AB - This study used archival data on a sample of 186,492 referrals from a southwestern state Juvenile Probation Commission to compare the characteristics of 5,439 male Black, Hispanic, and White juveniles with sexual behavior problems on the five most common sexual offenses in the data set. The characteristics of 181,053 juveniles of the three races without sexual behavior problems were also compared on the basis of the seven most common nonsexual offenses. The bases of comparison were the seven variables: reported incidence of sexual offenses, the primary caregivers or living arrangements, age, suspected sexual abuse, suspected emotional abuse, suspected physical abuse, and special education status, on which racial differences were found. Prevention and treatment implications of findings are discussed. PMID- 19306205 TI - Risk assessment of sexual offenders for extended supervision orders in New Zealand: basic principles and current practice. AB - A growing number of jurisdictions in North America, the United Kingdom, and Australasia have enacted legislation allowing for special sentencing, civil commitment, and community supervision options for high risk sexual offenders. In New Zealand, one example of this concern for public protection is the Parole (Extended Supervision) Amendment Act 2004, which provides for additional supervision of sexual offenders with child victims for up to 10 years after their release from prison. Recent experience with expert evidence and judicial decision making in such cases suggests that those involved in the process might benefit from a more thorough understanding of the current state of sexual offender risk assessment that can be provided by mental health professionals. PMID- 19306206 TI - Jury selection in child sex abuse trials: a case analysis. AB - Child sex abuse cases have been the target of considerable psycho-legal research. The present paper offers an analysis of psychological constructs for jury selection in child sex abuse cases from the defense perspective. The authors specifically delineate general and case-specific jury selection variables. General variables include authoritarianism, dogmatism, need for cognition, pretrial knowledge, and race/socioeconomic status. Case-specific variables include sexual attitudes, homonegativity, juror abuse history, and beliefs about children. The paper also provides a factual background of a representative case, incorporates relevant case law, identifies sources for voir dire and juror questionnaire items, and discusses lessons from the primary author's first experience as a trial consultant for the defense. PMID- 19306207 TI - Beyond Munchausen by proxy: a proposed conceptualization for cases of recurring, unsubstantiated sexual abuse allegations. AB - In the emerging literature, cases involving recurring, unsubstantiated allegations of child sexual abuse have generally been categorized as Munchausen by proxy. Recent scholars have recommended restricting the label to the original conceptualization, involving purposeful deception motivated by psychological needs for medical attention. This leaves many cases unclassified that do not fit the Munchausen by proxy criteria, involve significant risks to the child, and ultimately fall outside of existing structures for Child Protective Services/legal intervention. This paper presents a reconceptualization of such cases, proposing to label them "recurring sexual abuse allegation" cases. Defining the set of cases more clearly can aid child protection workers in their management and encourage research on prevalence, consequences to children, treatment strategies, and needed legal reforms. PMID- 19306208 TI - Errors of logic and scholarship concerning dissociative identity disorder. AB - The author reviewed a two-part critique of dissociative identity disorder published in the Canadian Journal of Psychiatry. The two papers contain errors of logic and scholarship. Contrary to the conclusions in the critique, dissociative identity disorder has established diagnostic reliability and concurrent validity, the trauma histories of affected individuals can be corroborated, and the existing prospective treatment outcome literature demonstrates improvement in individuals receiving psychotherapy for the disorder. The available evidence supports the inclusion of dissociative identity disorder in future editions of the Diagnostic and Statistical Manual of Mental Disorders. PMID- 19306209 TI - Synaptic organization of lobula plate tangential cells in Drosophila: Dalpha7 cholinergic receptors. AB - The nervous system of seeing animals derives information about optic flow in two subsequent steps. First, local motion vectors are calculated from moving retinal images, and second, the spatial distribution of these vectors is analyzed on the dendrites of large downstream neurons. In dipteran flies, this second step relies on a set of motion-sensitive lobula plate tangential cells (LPTCs), which have been studied in great detail in large fly species. Yet, studies on neurons that convey information to LPTCs and neuroanatomical investigations that enable a mechanistic understanding of the underlying dendritic computations in LPTCs are rare. We investigated the subcellular distribution of nicotinic acetylcholine receptors (nAChRs) on two sets of LPTCs: vertical system (VS) and horizontal system (HS) cells in Drosophila melanogaster. In this paper, we describe that both cell types express Dalpha7-type nAChR subunits specifically on higher order dendritic branches, similar to the expression of gamma aminobutyric acid (GABA) receptors. These findings support a model in which directional selectivity of LPTCs is achieved by the dendritic integration of excitatory, cholinergic, and inhibitory GABA-ergic input from local motion detectors with opposite preferred direction. Nonetheless, whole-cell recordings in mutant flies without Dalpha7 nAChRs revealed that direction selectivity of VS and HS cells is largely retained. In addition, mutant LPTCs were responsive to acetylcholine and remaining nAChR receptors were labeled by alpha-bungarotoxin. These results in LPTCs with genetically manipulated excitatory input synapses suggest a robust cellular implementation of dendritic processing that warrants direction selectivity. The underlying mechanism that ensures appropriate nAChR-mediated synaptic currents and the functional implications of separate sets or heteromultimeric nAChRs can now be addressed in this system. PMID- 19306211 TI - Mutations affecting the cAMP transduction pathway disrupt the centrophobism behavior. AB - Like vertebrates, invertebrates such as Drosophila display complex integrated behaviors that rely on locomotion for their execution. The use of genetic tools combined with sophisticated behavioral analysis has permitted researchers to investigate the brain structures implicated in those complex behaviors, such as locomotor activity. The video-tracking paradigm has allowed the study of multiple parameters of locomotor activity and has revealed that Drosophila exhibits centrophobism, a behavior related to spatial orientation. A structure/function study has demonstrated that the mushroom bodies (MBs) are implicated in this behavior. In the continuity of these former studies, we have investigated the role of the cAMP transduction pathway known to be implicated in olfactory learning and memory within the MBs. Here, we report that disturbing this pathway by using different mutants, such as dnc, rut, PKA, or amn, lead to centrophobism defect. Moreover, we found that the P[GAL4]C316 flies, used to rescue the amn mutant phenotype, like those previously reported for the learning and memory defect, are severely disturbed in centrophobism behavior. Remarkably, those flies are perfectly randomly distributed in the arena, suggesting that C316 flies carry an important mutated-gene implicated in neuronal networks required for proper spatial orientation. PMID- 19306213 TI - Assessment of exposure to pyrethroids and pyrethrins in a rural population of the Monteregie area, Quebec, Canada. AB - Pesticide use remains a preoccupation of the population and public health authorities given its possible impact on health. Pyrethroids can be listed among the widely used pesticides. The general population is potentially chronically exposed to pyrethroids mainly through food intake, but acute or sporadic exposures can also occur by other routes. Although pyrethroids are considered among the least toxic pesticides, their neurotoxic properties can affect humans, but current exposure levels in the population of Quebec are not known. The study thus aimed at assessing pyrethroid exposure in a rural, agricultural population during summer through measurements of urinary biomarkers. A total of 163 volunteers, 49 children and 114 adults, living in the Monteregie area of Quebec, participated in the study, which took place from June to August 2006, the period of intensive application of pesticides. Participants were asked to collect all their micturitions from 6 p.m. until the next morning, including first morning void, and to fill out a questionnaire to document factors that could potentially contribute to exposure. A gas-chromatography mass-spectrometry method was used to quantify six urinary metabolites resulting from pyrethroid biotransformation: cis and trans-2,2-(dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (cDCCA and tDCCA), 3-phenoxybenzoic acid (PBA), chrysanthemum dicarboxylic acid (CDCA), cis-2,2-(dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid (DBCA) and 4 fluoro-3-phenoxybenzoic acid (FPBA). Distributions of amounts of the six metabolites excreted per unit of body weight, during a standardized 12-hr period, followed the same decreasing pattern in adults and in children: tDCCA > PBA > cDCCA > CDCA > DBCA > FPBA. No statistically significant difference was observed between the two groups, but amounts of metabolites varied greatly among individuals, suggesting important interindividual variations in the absorbed doses of these compounds. No consistent associations were observed between the excretion of correlated metabolites and the various factors documented by questionnaire (personal factors, life habits, sources of exposure). Comparison of the current data with those observed in an urban population of the same province during the summer of 2005 suggests a greater summertime exposure to some pyrethroids in the rural population. PMID- 19306214 TI - Evaluation of oral malodour in left- and right-handed individuals. AB - Halitosis is a common social problem in the worldwide population. It is frequently associated with oral hygiene performance, so it may depend on the hand skill and cognitive ability of individuals. This study aimed to determine the relationship between handedness and pathologic oral halitosis. The study included a total of 1510 participants, and daily oral hygiene activities, hand preference, VSCs scores (volatile sulphur compounds) and clinical oral indices such as plaque index (PI) and tongue coating index (TCI) of each individual were recorded. Results of this study showed that there were no statistically significant differences between right- and left-handers (Groups I and II, respectively) according to age and oral hygiene practices. But it was found that PI and TCI were statistically higher in Group I with respect to Group II (Table 1). VSC scores, PI, and TCI were statistically higher in men compared to women (p<.01) (Table 2). There were statistically significant differences between women and men in the right-handed participant group (p<.01), while there were no statistically significant differences among the left-handers (p> .05) (Figure 1). Furthermore, statistically significant correlations were found between VSC scores and individual oral hygiene indices (p<.01) (Table 3). The lower halimetric values and clinical indices in left-handed individuals may be related with their oral healthcare and daily brushing habits. But unless detailed neurological investigations are undertaken, it cannot be suggested that the left-handers have better coordination and better-developed spatial abilities in oral hygiene procedures than the right-handers. PMID- 19306215 TI - Congenital candidiasis: confirmation of mother-neonate transmission using molecular analysis techniques. AB - We describe a case of congenital acquired candidiasis in a preterm female delivered through Caesarean section due to the premature rupture of the amniotic membrane. The neonate presented with suspected chorioamnionitis and erythematous desquamative skin. Candida albicans was isolated from the placenta, mouth, groin, and periumbilical lesions. The infant developed candidemia due to Candida albicans and the same yeast was also isolated from a catheter. Culture inoculated with swabs from the mouth and vagina of the mother yielded C. albicans and C. krusei. All C. albicans isolates from the mother and the neonate were visually indistinguishable by molecular typing techniques which included chromosomal karyotyping and restriction endonuclease analysis followed by pulsed-field gel electrophoresis. These findings allowed the clinical condition to be confirmed as congenital acquisition of candidiasis in this case. PMID- 19306217 TI - Cryptococcosis in domestic animals in Western Australia: a retrospective study from 1995-2006. AB - A retrospective study of cryptococcosis in domestic animals residing in Western Australia was conducted over an 11-year-period (from 1995 to 2006) by searching the data base of Murdoch University Veterinary Teaching hospital and the largest private clinical pathology laboratory in Perth. Cryptococcosis was identified in 155 animals: 72 cats, 57 dogs, 20 horses, three alpacas, two ferrets and a sheep. There was no seasonal trend apparent from the dates of diagnosis. Taking into account the commonness of accessions to Murdoch University, cats were five to six times more likely to develop this disease than dogs, and three times more likely than horses, while horses were almost twice as likely as dogs to become infected. Amongst the feline cohort, Ragdoll and Birman breeds were over-represented, while in dogs several pedigree breeds were similarly overrepresented. Dogs and horses tended to develop disease at an early age (one to five years), while cats were presented over a much wider range of ages. In cats and dogs the upper respiratory tract was the most common primary site of infection, while horses and alpacas tended to have lower respiratory involvement. The most striking finding of the study was the high frequency with which C. gattii was identified, with infections attributable to this species comprising 5/9 cats, 11/22 dogs, 9/9 horses and 1/1 alpaca, where appropriate testing was conducted. Preliminary molecular genotyping suggested that most of the C. gattii infections in domestic animals (9/9 cases) were of the VGII genotype. This contrasts the situation on the eastern seaboard of Australia, where disease attributable to C. gattii is less common and mainly due to the VGI genotype. C. gattii therefore appears to be an important cause of cryptococcosis in Western Australia. PMID- 19306219 TI - Media exposure and marijuana and alcohol use among adolescents. AB - We aimed to determine which media exposures are most strongly associated with marijuana and alcohol use among adolescents. In 2004, we surveyed 1,211 students at a large high school in suburban Pittsburgh regarding substance use, exposure to entertainment media, and covariates. Of the respondents, 52% were female, 8% were non-White, 27% reported smoking marijuana, and 60% reported using alcohol. They reported average exposure to 8.6 hr of media daily. In adjusted models, exposure to music was independently associated with marijuana use, but exposure to movies was independently associated with alcohol use. Implications, limitations, and suggestions for further research are discussed. PMID- 19306220 TI - Relationship violence and frequency of intoxication among low-income urban women. AB - Using data from the Welfare, Children, and Families project (1999), a probability sample of 2,280 low-income women with children living in low-income neighborhoods in Boston, Chicago, and San Antonio, we examine the effects of relationship violence before age 18 and in the past year on frequency of adult intoxication. Results obtained from a series of ordered logistic regression models suggest that sexual coercion before age 18 and minor and severe physical assault in the past year are independently associated with greater frequency of intoxication, net of a range of sociodemographic controls. The study's limitations are noted. PMID- 19306221 TI - Sociosexually unrestricted parents have more sons: a further application of the generalized Trivers-Willard hypothesis (gTWH). AB - BACKGROUND: The generalized Trivers-Willard hypothesis (gTWH) proposes that parents who possess any heritable trait which increases male reproductive success at a greater rate than female reproductive success in a given environment will have a higher-than-expected offspring sex ratio, and parents who possess any heritable trait which increases the female reproductive success at a greater rate than male reproductive success in a given environment will have a lower-than expected offspring sex ratio. AIM: One heritable trait which increases the reproductive success of sons much more than that of daughters is unrestricted sociosexual orientation. We therefore predict that parents with unrestricted sociosexual orientation (measured by the number of sexual partners, frequency of sex, and attitudes toward relationship commitment and sexual exclusivity) have a higher-than-expected offspring sex ratio (more sons). SUBJECTS AND METHOD: We analyse the US General Social Surveys and the National Longitudinal Study of Adolescent Health (Add Health), both with large nationally representative samples. RESULTS: Our analyses support the prediction from the gTWH. CONCLUSION: One standard deviation increase in unrestrictedness of sociosexual orientation increases the odds of having a son by 12-19% in the representative American samples. PMID- 19306222 TI - Use of chromomycin A3 staining in bovine sperm cells for detection of protamine deficiency. AB - Sperm chromatin integrity is essential for accurate transmission of male genetic information, and normal sperm chromatin structure is important for fertilization. Protamine is a nuclear protein that plays a key role in sperm DNA integrity, because it is responsible for sperm DNA stability and packing until the paternal genome is delivered into the oocyte during fertilization. Our aim was to investigate protamine deficiency in sperm cells of Bos indicus bulls (Nelore) using chromomycin A3 (CMA3) staining. Frozen semen from 14 bulls were thawed, then fixed in Carnoy's solution. Smears were prepared and analyzed by microscopy. As a positive control of CMA3 staining, sperm from one bull was subjected to deprotamination of nuclei. The percentage of CMA3-positive bovine sperm did not vary among batches. Only two bulls showed a higher percentage of CMA3-positive sperm cells compared to the others. CMA3 is a simple and useful tool for detecting sperm protamine deficiency in bulls. PMID- 19306223 TI - Evaluation of dietary status with respect to trace element intake from dry fruits consumed in Pakistan: a study using instrumental neutron activation analysis. AB - Instrumental neutron activation analysis has been utilized for the quantification of 24 major, minor and trace elements in dried apricots (Prunus armeniaca), dates (Phoenix dectylifera), figs (Ficus carica) and raisins (Vitis vinifera) using a low-power reactor as the neutron source. The weekly intakes of aluminium, arsenic, boron, calcium, chlorine, cobalt, chromium, iron, mercury, potassium, sodium, magnesium, manganese, molybdenum, antimony, selenium and zinc have been calculated and compared with the Recommended Dietary Allowance/Estimated Safe and Adequate Dietary Intake values through consumption of 100 g dried fruit. Fairly adequate levels of calcium, cobalt, iron, potassium and magnesium were determined in these dried fruits while the contribution of chlorine, chromium, molybdenum and sodium to the Recommended Dietary Allowance was found to be insignificant. Apricot is a nutritionally rich source of potassium, magnesium and cobalt, while chlorine and manganese contents were found to be highest in dates. Figs are an ample source of calcium, iron, magnesium and manganese, whereas raisins provide the highest intake of chromium, molybdenum and sodium. The toxic element contents of these dry fruits are considerably below the tolerance limits. PMID- 19306224 TI - Effectiveness of whole grain consumption in the prevention of colorectal cancer: meta-analysis of cohort studies. AB - The present work aimed to evaluate the effectiveness of whole grain consumption in preventing colorectal cancer. A systematic review with meta-analysis of 11 cohort studies was carried out. The age group of the population studied (1,719,590 participants) was between 25 and 76 years of age. The review evaluated the relative risks with the Cox proportional hazard model. The period of study varied from 6 to 16 years, where 7,745 persons developed colorectal cancer during the follow-up period. In the multivariate analysis, the highest quintile relative risk was 0.94 (95% confidence interval, 0.85-1.03), whereas that for the lowest quintile was 0.96 (95% confidence interval, 0.88-1.04). The location of tumors was also evaluated, with tumors in the colon demonstrating a relative risk of 0.93 (95% confidence interval, 0.83-1.02) and tumors in the recto a relative risk equal to 0.89 (95% confidence interval, 0.79-1.00). In this multivariate analysis, consumption of whole grains was inversely associated with the risk of developing colorectal cancer. PMID- 19306225 TI - Serum vitamin A in mothers and newborns in the city of Rio de Janeiro. AB - Retinol and carotenoid levels were determined spectrophotometrically in the blood of 222 mothers at delivery and in the umbilical cord of the newborns. We observed an important prevalence of vitamin A deficiency (VAD) among mothers and newborns (25.4% and 46.2%, respectively) as well as carotenoid inadequacy (52.3% and 92.6% in mothers and newborns, respectively), and an increased risk of VAD development among newborns whose mothers had VAD (odds ratio = 4.79). We found an increased risk of carotenoid inadequacy when VAD was already present in both groups (odds ratio = 2.21 and odds ratio = 6.85 in mothers and newborns, respectively). There were relationships between previous abortion and maternal VAD (P=0.022) and lower carotenoid levels among newborns (P=0.019), as well as inadequate maternal serum carotenoid levels and interdelivery interval less than 18 months. These findings suggest the need for interventional actions to prevent retinol and carotenoid inadequacy, and low antioxidant reserves. PMID- 19306226 TI - Invasion of the alveolar-capillary barrier by Aspergillus spp.: therapeutic and diagnostic implications for immunocompromised patients with invasive pulmonary aspergillosis. AB - Aspergillus spp. are leading causes of morbidity and mortality in immunocompromised patients. This review provides an overview of host defenses, the mechanisms by which the alveolar-capillary barrier is breeched by Aspergillus, and the implications for diagnosis and treatment of patients with invasive pulmonary aspergillosis. PMID- 19306227 TI - Quantification of the cell-substratum contact and cell lift-off under different intra/extracellular conditions. AB - Analyses of images of the cell-to-substratum region of contact have been carried out by the means of total internal reflection fluorescence (TIRF) microscopy during both the formation and the dissolution of cellular contacts. The evolutions of the cellular contacts are visualized during the adhesion process under normal and virus-infected intracellular conditions, and during the lift-off process under various toxicities of the extracellular medium fluid. Then, propositions are developed for quantifying the cell viabilities by estimating the increase in the area of contact for the adhesion process and by specifying the maximum intensity of the TIRF image for the lift-off process. PMID- 19306228 TI - White matter tract injury and cognitive impairment in human immunodeficiency virus-infected individuals. AB - Approximately half of those infected with the human immunodeficiency virus (HIV) exhibit cognitive impairment, which has been related to cerebral white matter damage. Despite the effectiveness of antiretroviral treatment, cognitive impairment remains common even in individuals with undetectable viral loads. One explanation for this may be subtherapeutic concentrations of some antiretrovirals in the central nervous system (CNS). We utilized diffusion tensor imaging and a comprehensive neuropsychological evaluation to investigate the relationship of white matter integrity to cognitive impairment and antiretroviral treatment variables. Participants included 39 HIV-infected individuals (49% with acquired immunodeficiency syndrome [AIDS]; mean CD4 = 529) and 25 seronegative subjects. Diffusion tensor imaging indices were mapped onto a common whole-brain white matter tract skeleton, allowing between-subject voxelwise comparisons. The total HIV-infected group exhibited abnormal white matter in the internal capsule, inferior longitudinal fasciculus, and optic radiation; whereas those with AIDS exhibited more widespread damage, including in the internal capsule and the corpus callosum. Cognitive impairment in the HIV-infected group was related to white matter injury in the internal capsule, corpus callosum, and superior longitudinal fasciculus. White matter injury was not found to be associated with HIV viral load or estimated CNS penetration of antiretrovirals. Diffusion tensor imaging was useful in identifying changes in white matter tracts associated with more advanced HIV infection. Relationships between diffusion alterations in specific white matter tracts and cognitive impairment support the potential utility of diffusion tensor imaging in examining the anatomical underpinnings of HIV-related cognitive impairment. The study also confirms that CNS injury is evident in persons infected with HIV despite effective antiretroviral treatment. PMID- 19306229 TI - Acquired immunodeficiency syndrome and the blood-brain barrier. AB - The blood-brain barrier (BBB) plays a critical role in normal physiology of the central nervous system by regulating what reaches the brain from the periphery. The BBB also plays a major role in neurologic disease including neuropathologic sequelae associated with infection by human immunodeficiency virus (HIV) in humans and the closely related simian immunodeficiency virus (SIV) in macaques. In this review, we provide an overview of the function, structure, and components of the BBB, followed by a more detailed discussion of the subcellular structures and regulation of the tight junction. We then discuss the ways in which HIV/SIV affects the BBB, largely through infection of monocytes/macrophages, and how infected macrophages crossing the BBB ultimately results in breakdown of the barrier. PMID- 19306232 TI - Free radical generation in rosmarinic acid investigated by electron paramagnetic resonance spectroscopy. AB - Free radical generation as a result of oxidation reactions of rosmarinic acid (RA), a caffeic acid ester with 3,4-dihydroxyphenyllactate, was investigated by electron paramagnetic resonance (EPR) spectroscopy using a variety of oxidation conditions. Limitations and possibilities of using the various methodologies to obtain information about the reaction chemistry of polyphenols are discussed. Three different spectra were detected dependent on the pH and oxidizing agent. Feasible structures for the observed radicals were tested by density functional theory (DFT) calculations and the results indicate that oxidation reactions can occur at both of the catechol groups. PMID- 19306230 TI - Lithium therapy for human immunodeficiency virus type 1-associated neurocognitive impairment. AB - The objective of this study was to assess lithium safety and tolerability and to explore its impact on cognition, function, and neuroimaging biomarkers in human immunodeficiency virus (HIV)-infected subjects with cognitive impairment. Fifteen cognitively impaired HIV-infected subjects were enrolled in this 10-week open label study of lithium 300 mg twice daily. Neuroimaging was performed at baseline and following 10 weeks of treatment and included magnetic resonance spectroscopy (MRS), diffusion tensor imaging (DTI), and functional MRI (fMRI). Thirteen of the 14 subjects (93%) that complied with the study visits were able to complete the study on lithium and 11 out of 13 (79%) completed the study at the originally assigned dose of 300 mg twice daily. There were no significant changes in CD4(+) lymphocyte cell count and plasma HIV RNA. Cognitive performance and depressive mood did not improve significantly after the 10-week lithium treatment; however, neuroimaging revealed a decrease in the glutamate+glutamine (Glx) peak in the frontal gray matter, increased fractional anisotropy, and decreased mean diffusivity in several brain areas, and changes in brain activation patterns, suggestive of improvement. These results suggest that lithium can be used safely in HIV-infected individuals with cognitive impairment. Furthermore, the neuroimaging results suggest that lithium may improve HIV-associated central nervous system (CNS) injury; thus, further investigations of lithium as an adjunctive treatment for HIV-associated cognitive impairment are warranted. PMID- 19306233 TI - Educational skills: long-term outcome and predictors following paediatric traumatic brain injury. AB - Given that reading, spelling and arithmetic skills are acquired through childhood, their development may be compromised following a childhood traumatic brain injury (TBI). The present study examined educational skills (reading accuracy, spelling and arithmetic) at a mean follow-up interval of 6.8 years post injury in children who had sustained a mild, moderate, or severe TBI at two ages: 'Young' (age at injury: 3-7 years, n = 48) and 'Old': (age at injury: 8-12 years, n = 36). Comparisons between the young and old TBI groups resulted in inconsistent findings. While a dose-response relationship for severity was evident for the young group, this was not always the case for the old group. Significant predictors of outcome included both severity and acute intellectual function. PMID- 19306234 TI - [Clinical effects of the therapy method deep oscillation in treatment of sports injuries]. AB - BACKGROUND: In Germany approximately 2 million sports injuries occur per year. Most common are distorsions and ligamentous injury going along with posttraumatic lymphoedema. With the therapy method deep oscillation very good results in other indications and in lymphdrainage were reached. The purpose of this experimental study was the evaluation of the effects of the therapy method deep oscillation in immediate therapy and after-care of different sports injuries in addition to usual care (complex physical and medical therapy). METHODS: Two soccer teams were supported by a sports medicine section of a rehabilitation hospital. In n = 14 people (mean age 23.9 years) 49 sports injuries of different kind were treated. RESULTS: Subjective rating of the symptoms by VAS improved significant (p = 0.001) from 8.7 (baseline) to 2.1 points (post-treatment). Objective rating by the attending physician according to different clinically relevant parameters lead to "very good" or "good" results in 90 % of the patients. CONCLUSION: In conclusion it can be stated that therapy method deep oscillation is an easy to use and comparably cost effective adjuvant therapy option. We already had good experience with it in other indications concerning reabsorption of oedema, reducing pain, anti-inflammatory effect, promotion of motoricity, promotion of wound healing, anti-fibrotic effect and improvement in trophicity and quality of the tissue. All these mentioned effects can be confirmed in the treatment of patients with acute sports injury and trauma. The soft mode of action is the reason that in contrast to other electric and mechanical therapies it is no contraindication in immediate therapy. In general we noted no side effects; patients were highly compliant and rated this therapy as very good. PMID- 19306235 TI - [Ambulatory CT-assisted thoracic sympathetic block as an additional approach to treatment of complex regional pain syndromes after sport injuries]. AB - AIM: Also after bagatelle trauma, some patients can develop a complex regional pain syndrome (CRPS). The limb concerned usually shows a marked temperature difference to the contralateral side. Apart from a doughy swelling, the patient is impaired in particular by burning pain and a restricted range of motion. The objective of the present study was to examine the influence of early thoracic sympathetic nerve blockade on the basis of clinical and quantified blood flow parameters. METHODS: In 7 young patients (average age 17.7 [15 - 21] years) with a clinically developing type 1 CRPS after sport injuries (mountain bike, bike, skateboard, hockey and go-kart accidents), occult injury and ligament lesions were ruled out using conventional imaging and MRI. In addition to pain-adapted physiotherapy and pharmacotherapy with analgesics and calcitonin, sympathetic nerve blockade was performed three times at two-day intervals. The CT-assisted puncture was performed in the prone position at the level of the intervertebral space of the 2nd/3 rd thoracic vertebral bodies. In the correct paravertebral position, a 10-cm long, 22 G Seibel-Gronemeier needle was used for the successive injection of 6 - 10 ml Carbostesin 0.5 %. By adding a small amount of contrast medium, it was possible to visualise clearly the distribution of the sympathicolytic agent in the control scan in each case. Before and after the intervention, colour-coded duplex sonography (CCDS) of the affected limb arteries was performed on all patients. RESULTS: The injection needle was correctly placed in all patients, without complications. The medication mixture was observed to have distributed properly. After the intervention, all patients reported the immediate onset of marked pain relief, whereby they recovered fully over the further course. The increase in peripheral blood flow was shown by a significant improvement in flow in the CCDS. CONCLUSION: Outpatient CT-assisted temporary sympathetic nerve blockade is an effective and low-complication therapeutic option for the supportive treatment of patients with CRPS. PMID- 19306236 TI - [Safety requirements and risk factors of skiers and snowboarders]. AB - BACKGROUND: Injuries in leisure sports, especially with skiers and snowboarders are a challenge for medicine as well as for economy. The risk of getting injured depends not only on the performed sport but also on the individual readiness to take a risk and the worn safety devices. MATERIAL AND METHOD: 663 persons performing leisure sports in an alpine ski resort were questioned concerning their individual readiness to take a risk and need for security. It was distinguished between skiers and snowboarders. Also the worn safety devices as well as the reasons for and against the use of safety devices were asked. As safety devices helmets, back protectors, wrist guards, knee-, shin- and elbow pads were considered. There were also questions about the behaviour on the ski slope and injuries so far. RESULTS: 52.5 % of all questioned persons are skiers, 36.0 % snowboarders, 11.5 % practise both sports. Safety devices are widely accepted in leisure sports: 78.1 % of all persons questioned wear at least one device. 76.8 % wear it for their own safety. The most common reason against safety devices are "lack of need" with 59.6 %. 28 % mentioned lack of comfort. Males show a significantly riskier behaviour on ski piste than females, 7 versus 5 on an analogue scale 0 - 10. After all 47.2 % of all persons have suffered from an injury, which needed medical treatment. In 75.7 % the cause was a fall, 14,7 % were results of a collision. In this context 49.5 % would appreciate the introduction of a ski slope controller. 90.5 % deny the consumption of alcohol while performing their sport and 78.4 % consider the consumption of alcohol as a risk either for themselves or for others. CONCLUSION: Our study shows a broad acceptance of safety devices in leisure sport. A lot of studies have proved the value of helmets to prevent injuries. The introduction of mandatory helmets at least for children and juveniles in alpine sports would be well accepted. Males show a significantly riskier behaviour on ski pistes than females. Efficient controls on ski pistes to avoid ruthless behaviour would be appreciated and declined by equal numbers. PMID- 19306237 TI - [ACL reconstruction in skeletally immature patients]. AB - AIM: The treatment of choice for skeletally immature patients with anterior cruciate ligament (ACL) deficiency is an ACL reconstruction. In a retrospective study we analysed the clinical results after ACL reconstruction in skeletally immature patients. MATERIAL AND METHOD: In 55 immature patients the ACL was replaced with a four strand-hamstring graft in an anatomic transepiphyseal origin. The fixation was by means of an extracortical button femoral and a suture washer or staple tibial. Patient mean age at operation was 13 years (8 to 16 years). RESULTS: The mean follow-up was at 3.2 years (1 to 7.5 years) postoperatively. The objective IKDC 2000 score was 90.7 % normal or almost normal values. The mean Lysholm score was 94.1 points (70 - 100), the mean Cincinnati knee score was 93.9 points (76 - 100) and 88 % of the patients went back to normal or almost normal sports according to the Tegner score. Stability testing performed with the KT-1000 arthrometer was 1.0 mm (0 - 4 mm) (preoperative average 5.8 mm). The traumatic re-rupture rate was 5.5 %. In two additional cases a partial rupture of the graft was analysed by arthroscopy. Growth deformities or leg length differences were not seen in any case. CONCLUSION: ACL reconstruction with hamstrings and extracortical fixation showed good results and might be the treatment of choice in immature patients. The complication rate was low and there were no postoperative growth deformities. PMID- 19306238 TI - [Breakdancer's "Headspin Hole" - first description of a common overuse syndrome]. AB - INTRODUCTION: Several typical injuries due to breakdancing have been described in medical literature, but until now the "headspin hole" has not been mentioned. METHODS: We conducted internet research and interviewed 106 breakdancers. RESULTS: 60.4 % of the breakdancers suffered from overuse of the scalp due to the headspin. Loss of hair was found in 31.1 % and painless lumps on the head in 23.6 %. Inflammation of the scalp was reported in 36.8 %. DISCUSSION: The "Headspin Hole" is a common overuse syndrome of the scalp specific to headspins causing hair loss, inflammation and lumps on the skull. PMID- 19306239 TI - [Dose reduction in high-resolution MSCT examinations of the chest for early detection of pneumonia in immunocompromised patients]. AB - PURPOSE: The purpose of this study was to optimize high-resolution MSCT chest protocols for the evaluation of symptomatic immunosuppressed patients with suspected pneumonia using a dose-simulating program. MATERIALS AND METHODS: Using the MSCT (Siemens, Erlangen, Germany), 30 immunosuppressed patients with suspected pneumonia were examined with a low-dose HRCT of the chest (120 kV, 100 eff.mAs and collimation of 4 x 1 mm). A dose-simulating program was used to reconstruct the raw data at four different dose levels (70, 50, 35 und 25 mAs). For dose simulation for each mAs product, the correspondent noise level was added to the data. Images were generated with a slice thickness of 1 mm and 5 mm in the lung window. The images were then evaluated independently by two radiologists and graded on a scale of 1 to 3 points: 1 = no pneumonia, 2 = unclear, 3 = pneumonia. A receiver operating curve (ROC) analysis was performed to calculate the area under the curve (AUC). The actual dosage in mSv was calculated. The sensitivity and specificity were evaluated. RESULTS: Out of 30 patients, 7 had a normal chest finding and 23 had pneumonia. The area under the ROC curve (AUC) was 1.0 for every dosage and slice thickness. Infiltrates were detected correctly for all dosage levels. The sensitivity was 100 % for all dose levels and slice thicknesses. There was one false positive finding at 35 mAs and 1 mm slice thickness. At this dose level the specificity was reduced to 93 %. A reduction to 25 mAs had no influence on the detection of pneumonia. Thus, MSCT examinations of the chest can be performed with 25 mAs without missing the right diagnosis, resulting in an effective dose of 1.15 mSv (men), 1.5 mSv (women) and a CTDIvol of 2.5 mGy. CONCLUSION: For the evaluation of pneumonia in immunocompromised patients, MSCT examination of the chest can be performed with 25 mAs. Thus, radiation exposure was reduced to a quarter compared to the standard protocol. PMID- 19306240 TI - [The value of a standardized ultrasound in patients with fever]. AB - PURPOSE: Transcutaneous ultrasound is not a common imaging procedure in patients with fever. The aim of this study is to demonstrate the diagnostic value of a complete standardized ultrasound examination of patients with fever. MATERIALS AND METHODS: From March 2003 to October 2003, we examined 200 patients with fever using ultrasound. We performed a standardized ultrasound examination including the collar, thoracal, cardial, abdominal regions, and small parts/soft tissue regions depending on clinical symptoms. We retrospective analyzed the patient history regarding fever (origin known vs. unknown) and the diagnostic value of ultrasound (no ultrasound diagnosis vs. ultrasound diagnosis). RESULTS: At the time of inclusion in the study, the origin of fever was known in 124 of 200 patients (62 %) and unknown in 76 of 200 patients (38 %). Ultrasound did not result in a diagnosis in 86 of 200 patients (43 %). A pathological finding was detected in 114 of 200 cases (57 %) as the origin of the fever. New causes for fever were found in 24 of 200 patients (12 %) by means of ultrasound. The most common diagnoses (67 %) in this subgroup were abscesses and splenic infarcts. CONCLUSION: In the clinical course of patients with fever, ultrasound is a diagnostic tool for the documentation and exclusion of medical findings. With this study we were able to show that ultrasound results in the correct diagnosis in one third of patients with fever of an unknown origin and may provide important additional information in patients with fever. PMID- 19306241 TI - Developmental dysplasia of the hip: ultrasound screening and treatment. How are they related? AB - An assessment of the current clinical and ultrasound screening programmes. Early treatment by splintage is advocated for instability and Graf type III dysplasia of the hip. PMID- 19306242 TI - Septic arthritis of the hip - current concepts. AB - Septic arthritis of the hip is the commonest septic condition during growth, reaching a distinct peak in frequency during infancy. The aetiology is a haematogenous joint infection. Indicative signs are severe pain when moving the joint, septic appearance and a poor general condition of these small and young patients. The diagnosis often can be difficult in infants since septic temperatures are not always present. An ultrasound scan shows the hip joint effusion and the capsular distension. X-ray investigation helps to exclude defective situations. Therapeutic options are: in patients with short history without radiologically visible complications we recommend repeated arthroscopic irrigation and in patients with long history and a radiologically visible defect of the femoral head or dislocation we recommend arthrotomy and open revision or reduction of the hip joint. PMID- 19306243 TI - Slipped Upper Femoral Epiphysis (SUFE): to do or not to do in sufe. AB - Slipped upper femoral epiphysis (SUFE) is not an uncommon hip disease in adolescence. Usually, diagnosis of SUFE is delayed, has high risk of sequela and poor long-term results. This paper reviews the literature with a focus on epidemiology, aetiology, diagnosis, treatment and results regarding SUFE. PMID- 19306244 TI - Proximal focal femoral deficiency (PFFD): management options and controversies. AB - Proximal focal femoral deficiency (PFFD) is a rare congenital anomaly characterised by failure of normal development of the proximal femur and hip joint. Significant variability in the clinical presentation and degree of deficiency is common. Current management strategies aimed at improving functional ambulation are largely dependent on the degree of femoral shortening and the status of the hip and knee joint. Treatment of acetabular deficiency and proximal femoral deformity in cases of PFFD must be individualised. Reconstruction of the hip joint with pelvic and femoral osteotomies may be possible in mild cases of PFFD. Stability of the hip and knee joint must be achieved prior to consideration for limb lengthening strategies. Severe cases of PFFD may be beyond surgical correction and warrant alternative strategies such as rotationplasty or selective amputation to facilitate prosthetic fitting. PMID- 19306245 TI - The hip in osteochondrodysplasias: general rules for diagnosis and treatment. AB - The management of hip pathology in osteochondrodysplasia (skeletal dysplasia) is complex and a multidisciplinary approach is vital. Thorough clinical assessment and knowledge of the natural history of the different disorders provides the basis for this. PMID- 19306246 TI - Tumours and tumour-like lesions of the hip in the paediatric age: a review of the Rizzoli experience. AB - Bone tumours and tumour-like lesions of the hip in children are rare. Signs and symptoms of these tumours are generally nonspecific. Delay of diagnosis is not uncommon. A high index of suspicion in young patients presenting with persistent pain and without history of trauma, that is unresolved with conservative therapy should prompt further investigation, including radiographs or computed tomography scan of the pelvis. In the experience of the Istituto Rizzoli, in patients less than 14 years (mean 9 years, ranged from 6 months to 14 years), 752 tumours and tumours-like lesions occurred in the pelvis or proximal femur, involving the hip. Tumour-like lesions accounted for 322 cases (simple bone cyst in 255, eosinophilic granuloma in 43, aneurismal bone cyst in 34), benign tumours for 340 cases (osteoid osteoma in 229, fibrous dysplasia in 63, exostosis in 48) and malignant tumours for 80 cases (Ewing's sarcoma in 53 and osteosarcoma in 27). The epidemiology, pathology, clinical presentation, and radiograph findings are discussed for each of these tumours.Treatment of these tumours differs from observation or minimally invasive treatment for most pseudotumoural lesions, intralesional excision or termoablation for benign bone tumours and wide resection for malignant bone tumours. In this latter group, chemotherapy is required and often administered pre- and postoperatively. PMID- 19306247 TI - Strategies of hip management in neuromuscular disorders: Duchenne Muscular Dystrophy, Spinal Muscular Atrophy, Charcot-Marie-Tooth Disease and Arthrogryposis Multiplex Congenita. AB - Joint contractures, subluxation and dislocation are common problem in children with neuromuscular disorders. Medical, surgical and rehabilitative approaches can be used to maintain patient function and comfort. Contracture release, hip dysplasia correction and procedures to address or prevent hip subluxation or dislocation, are not always necessary since patients can be asymptomatic and surgical treatment will not always be successful in maintaining a reduced hip. In fact, controversy surrounds the management of hip disorder in children with Duchenne Muscular Dystrophy, Spinal Muscular Atrophy, Charcot-Marie-Tooth Disease and Arthrogryposis Multiplex Congenita. Patients with neuromuscular disorders also frequently develop a progressive scoliosis with pelvic obliquity which may affect sitting balance and become painful. Most subluxations and dislocations have the tendency to occur on the high side of a tilted pelvis. Spinal stabilisation is sometimes necessary to improve the pelvic tilt and to prevent further increase. The present article provides an overview of the current strategies of hip management in neuromuscular disorders. PMID- 19306248 TI - Strategies of hip management in myelomeningocele: to do or not to do. AB - Paralytic hip dislocation in myelomeningocele is common and can be a complicated problem. This review summarizes results of surgical techniques employed in this patient population in order to achieve and maintain reduction of paralytic hip dislocations. This review also examines the controversial question of whether or not hip surgery in patients with myelomeningocele provides improved functional results. Finally this paper suggest appropriate goals and recommendations for treatment of the paralytic hip dislocation in myelomeningocele. PMID- 19306249 TI - "Postural Management" to prevent hip dislocation in children with cerebral palsy. AB - Muscolo-skeletal disorders and hip deformity play a key role on future development of motor and adaptive functions in children with cerebral palsy (CP). Lateral migration of the femoral head increases on average 7.7% per year and may progress in association with acetabular dysplasia to hip dislocation. Conservative preservation of muscle length and balance may prevent or reduce femoral head migration and acetabular dysplasia. Equipment for position lying, sitting, and standing is an established method of maintaining muscle length and joint range. French approach (Le Metayer et al) include the use of customised plaster cast orthoses, for sitting and standing positions, called siege moule and gouttiere, tailor-made according to the child's muscolo-skeletal characteristics and motor abilities. At our department, hips with clinical and radiological evidence of displacement unresponsive to pharmacological and surgical treatment were treated with postural management. The two cases we show in this paper underwent a physiotherapy programme (neurodevelopment treatment) twice a week and a seated postural programme 5 hours a day with the siege moule. Hip radiographs were measured with the migration percentage (MP) described by Reimers. The follow up measurements showed a progressive reduction of MP values of the hip treated, confirming the significant benefit from the combined non surgical approach. The study supports the evidence that conservative management of hip deformity with siege moule can be successful if implemented before the development of hip dysplasia. PMID- 19306250 TI - Relationship between kinematic knee deviations and femoral anteversion in children with cerebral palsy. AB - The aim of the study was to determine the possible correlation between the degree of femoral anteversion and the quantitative data obtained by 3D Gait Analysis (GA) and then to investigate the relationship between femoral anteversion and the reduced knee flexion during swing phase in children with Cerebral Palsy. Twenty seven diplegic children with severe rectus femoris spasticity and 20 healthy children (CG) were considered. Clinical evaluation of femoral anteversion, Duncan Ely test and Gait Analysis were performed in all patients. From Gait Analysis data some indices were identified and calculated and statistical analysis performed. Clinical evaluations made the distinction between patients with excessive femoral anteversion (Group 1) and those with normal value (Group 2). Both groups showed a blunt maximum of knee flexion in swing (KMSw), representative of rectus femoris spasticity, but two different gait strategies were found for the timing of KMSw. Group 1 exhibited a reduced KMSw value with its timing close to normal value and an excessive hip internal rotation (Mean Hip Rotation index), correlated to high femoral anteversion; Group 2 presented a limited KMSw and a significant delay of its timing, with Mean Hip Rotation index close to Control Group. No differences were found for other indices. The results demonstrated that the presence of reduced KMSw only can be directly connected to excessive femoral anteversion; the coexistence of reduced KMSw and its delayed timing reveals that the rectus femoris spasticity may be due to rectus spasticity added to an incorrect motor selective control. The results are clinically crucial for treatment strategies (derotative femoral osteotomy vs rectus transfer). PMID- 19306251 TI - D.D.S.H.: developmental dysplasia of the spastic hip: strategies of management in cerebral palsy. A new suggestive algorithm. AB - Hip problems in cerebral palsy are relatively frequent (25-75%). Subluxation and dislocation of the hip is proportional to the neuromuscular involvement and is often due to alteration caused by spastic muscle forces acting on the femoral head in the acetabular cavity. The EMMA approach (Early Multilevel Minimally invasive Approach) has been designed to restore muscle balance, decrease hip migration and prevent bone deformities thereby avoiding future pain with minimal biological cost to the patient. EMMA is suitable for most patients, especially those with increased tone, poor muscle control and selectivity, Reimer Index (R.I.) 20%. We consider age and R.I crucial prerequisites for treatment steps. EMMA 1) age 2-4 years, RI 20%: multilevel injection of botulinum toxin in case of muscular hyperactivity without morphological alterations of the couple muscle tendon (contractures). EMMA 2) age 4-6, RI 20%: multilevel aponeurectomies in case of muscular hyperactivity with morphological alterations of the couple muscle-tendon (retraction). EMMA 3) early bone surgery (growth plates). This approach has been adopted in the last 4 years to prevent bone deformities and give early mobilisation and early control of the pain. EMMA is simple to apply even in infants, both for hip containment and to decrease spasticity. PMID- 19306252 TI - HPLC enantioseparation of beta2-homoamino acids using crown ether-based chiral stationary phase. AB - RP high-performance liquid chromatographic methods were developed for the enantioseparation of eleven unusual beta(2)-homoamino acids. The underivatized analytes were separated on a chiral stationary phase containing (+)-(18-crown-6) 2,3,11,12-tetracarboxylic acid as chiral selector. The effects of organic (alcoholic) and acidic modifiers, the mobile phase composition and temperature on the separation were investigated. The structures of the substituents in the alpha position of the analytes substantially influenced the retention and resolution. The elution sequence was determined in some cases: the S enantiomers eluted before the R enantiomers. PMID- 19306253 TI - HILIC separation and quantitation of water-soluble vitamins using diol column. AB - Hydrophilic interaction liquid-chromatography (HILIC) in conjunction with diode array detection has been applied for the separation of selected-water-soluble vitamins using an end-capped HILIC-diol column. Vitamins with significant biological importance, such as thiamine (B(1)), riboflavin (B(2)), nicotinic acid (B(3)), nicotinamide (B(3)), pyridoxine (B(6)), folic acid (B(9)), cyanocobalamin (B(12)) and ascorbic acid (vitamin C) were simultaneously separated. Chromatographic conditions including type and percentage of organic modifier in the mobile phase, pH, type and concentration of buffer salt and flow rate were investigated. ACN was shown to offer superior separation for the compounds tested as compared to methanol, isopropanol and THF. Isocratic separation and analysis were achieved for six vitamins (B(1), B(2), nicotinic acid/nicotinamide, B(6) and C) at ACN-H(2)O 90:10, containing ammonium acetate 10 mM, triethylamine 20 mM, pH 5.0, using a flow rate of 0.8 mL/min, while a gradient was necessary to resolve a mixture of all eight water-soluble vitamins. The HILIC method was validated and successfully applied to the analysis of a pharmaceutical formulation and an energy drink negating the need for time consuming clean-up steps. PMID- 19306254 TI - Simultaneous determination of seven fluoroquinolones in royal jelly by ultrasonic assisted extraction and liquid chromatography with fluorescence detection. AB - A method for the quantitative determination of seven fluoroquinolone antibacterial agents (FQs) used in beekeeping, viz. ciprofloxacin, norfloxacin, ofloxacin, pefloxacin, danofloxacin, enrofloxacin, and difloxacin, in royal jelly samples was developed on the basis of high performance liquid chromatography with fluorescence detection. Sample preparation included deproteination, ultrasonic assisted extraction with a mixed inorganic solution of monopotassium phosphate (KH(2)PO(4)) and ethylenediaminetetraacetic acid disodium salt (Na(2)EDTA), and clean-up on a solid-phase extraction cartridge. The extraction procedure was optimized with regard to the amount of inorganic solvent and the duration of sonication for royal jelly as a complicated matrix. Overall recoveries for FQs ranged from 85.9 to 99.1% for royal jelly with standard deviations between 2.79 and 6.27%. Limits of quantification were 2-40 ng/g for seven FQs in royal jelly. A total of 57 real royal jelly samples collected from beekeepers and supermarkets were analyzed. The three most abundant honeybee-use FQs, i. e. ofloxacin, ciprofloxacin, and norfloxacin, were determined in some royal jelly samples in concentrations ranging from 11.9 to 55.6 ng/g. Unexpectedly, however, difloxacin was found at concentrations of about 46.8 ng/g in one sample although it is rarely used in beekeeping. The presented method was successfully applied to quantify FQs in real royal jelly samples. PMID- 19306255 TI - Determination of thermodynamic pKa values of benzimidazole and benzimidazole derivatives by capillary electrophoresis. AB - Thermodynamic pK(a) values for benzimidazole and several substituted benzimidazoles were determined by CE. Electrophoretic mobilities of benzimidazoles were determined by CE at different pH levels and ionic strengths. The dependence of electrophoretic mobilities on pH was used to obtain pK(a) values at different ionic strengths. Extrapolations to zero ionic strength were used to determine the thermodynamic pK(a) values. Using this method the thermodynamic pK(a) values of 15 benzimidazoles were determined and found to range from 4.48 to 7.38. Results from the CE measurements were compared with spectrophotometric measurements which were evaluated at wavelengths where the highest absorbance difference for varying pH was recorded. The two analytical techniques were in good agreement. PMID- 19306258 TI - Tandem affinity purification of protein complexes in mouse embryonic stem cells using in vivo biotinylation. AB - In dissecting the pluripotent state in mouse embryonic stem (ES) cells, we have employed in vivo biotinylation of critical transcription factors for streptavidin affinity purification of protein complexes and constructed a protein-protein interaction network. This has facilitated discovery of novel pluripotency factors and a better understanding of stem cell pluripotency. Here we describe detailed procedures for in vivo biotinylation system setup in mouse ES cells, and affinity purification of multi-protein complexes using in vivo biotinylation. In addition, we present a protocol employing SDS-PAGE fractionation to reduce sample complexity prior to submission for mass spectrometry (MS) protein identification. PMID- 19306259 TI - Imaging neural stem cell fate in mouse model of glioma. AB - This unit describes a protocol for following the fate of stem cells in real time in a mouse model of glioma. Stem cells and tumor cells can be transduced with lentiviral vectors bearing two different luciferases, firefly luciferase (Fluc) and Renilla (Rluc) luciferase, respectively. With the cells labeled in this manner, bioluminescence imaging can be used to study the fate of stem cells in glioma-bearing brains in vivo. PMID- 19306260 TI - Interval approach to assessing antitumor activity for tumor xenograft studies. AB - In preclinical cancer drug screening tumor xenograft experiments, the tumor growth inhibition ratio (T/C) is commonly used to assess the antitumor activity of the agents. Unfortunately, this measurement can discard useful data and result in a high false-negative rate. Furthermore, the degree of antitumor activity based on the T/C ratio is assessed on the basis of an arbitrary cutoff point that does not reflect variations in different tumor lines. To overcome these drawbacks, we propose an adjusted area-under-the-curve (aAUC) ratio to quantify tumor growth inhibition. A nonparametric bootstrap t-interval of the aAUC ratio is also proposed for assessing the significance of the antitumor activity of the agents. The proposed method is then applied to a real tumor xenograft study. PMID- 19306261 TI - Sequencing of peptide-derived Amadori products by the electron capture dissociation method. AB - The electron capture dissociation (ECD) of peptide-derived Amadori products has been successfully applied for their sequencing. In contrast to the collision induced dissociation (CID), based on the vibrational excitation of peptides, the ECD method does not produce ions formed by fragmentation of the hexose moiety, that facilitates interpretation of the obtained spectra. The fragmentation spectrum is dominated by c(n) and z.(n) ions, providing the sufficient information for sequencing of peptides and establishing the location of glycated Lys residues in the peptide chain. The ECD experiments were conducted on a series of synthetic peptides and unseparated digests of glycated ubiquitin. PMID- 19306262 TI - Cell-patterning using poly (ethylene glycol)-modified magnetite nanoparticles. AB - Development of cell-patterning techniques is a major challenge for the construction of functional tissues and organs in tissue engineering. Recent progress in surface chemistry has enabled spatial control of cell adhesion onto cultural substrates by varying hydrophilicity, for example, by using poly (ethylene glycol) (PEG). In the present study, we developed a novel cell patterning procedure using PEG-modified magnetite particles (PEG-Mags) and magnetic force. Using an array-patterned magnet, PEG-Mags were magnetically patterned on the surface of a tissue culture dish. The resultant substrate surface consisted of two regions: the PEG-Mag surface that acts as a cell resistant region and the native substrate surface that promotes cell adhesion. When human keratinocyte HaCaT cells were seeded onto the PEG-Mag-patterned surface, cells adhered only to the native substrate surface, resulting in cell patterning on the tissue culture dish. The patterned PEG-Mags were then washed away to expose the native substrate surface, and thereafter, when mouse myoblast C2C12 cells were seeded to the dish, cells adhered to the exposed substrate surface, resulting in a patterned coculture of heterotypic cells. Moreover, it is worth noting that the magnetic force-based cell-patterning procedure is not limited by the property of cultural substrate surfaces, and that cell-patterning of mouse fibroblast NIH3T3 cells on a monolayer of HaCaT cells was successfully achieved using PEG-Mags and magnetic force. These results indicate that this procedure provides a novel concept for cell-patterning and may be useful for tissue engineering and cell biology. PMID- 19306263 TI - Different expression of NR2B and PSD-95 in rat hippocampal subregions during postnatal development. AB - The different expressions of NR2 and synaptic-associated proteins have been studied by protein and mRNA level with immunoblotting, in situ hybridization, or immunogold analysis. But the relationship between NR2 subunits and PSD-95 family proteins is still controversial. In this study, we used immunofluorescent staining to assess NR2B and PSD-95 expressions and the relationship between them in CA1, CA3, and DG of rat hippocampus on different postnatal day. In CA1, NR2B expression decreased with age. It was high at birth, reached a plateau at P4, and declined gradually after P7. In CA3, NR2B expression was similar to that in CA1. But the stratum lucidum was devoid of staining. In DG, the NR2B expression retained a higher level. From P0 to P2, the PSD-95 expression in CA1 increased gently, and then declined slightly. After P7, the PSD-95 expression increased sharply till P28, and decreased again. In CA3 and DG, the PSD-95 expression is very similar except that low-level of PSD-95 was found in the CA3 stratum lucidum. The expression of NR2B did not correlate with that of PSD-95 in CA1 and the DG granular and molecular layer. Only in CA3 and DG polymorphic layer, there was a negative correlation. The results suggested in hippocampal subregions, CA3 and DG may be more plastic than CA1.The NR2B and PSD-95 expression have distinct regional and cell specific distribution. The different regional distribution pattern may relate to the different physiological functions during postnatal development. PMID- 19306264 TI - Physical properties of the probe electrospray ionization (PESI) needle applied to the biological samples. AB - Probe electrospray ionization (PESI) is a recently developed technique that uses a fine solid needle as a probe for sampling biological materials. In this study, we quantified the volume of liquid sample picked up by the solid needle with the tip diameter of approximately 700 nm and the apex angle of approximately 60 degrees. The amounts of low-viscosity samples (rat urine) loaded on the tip of the needle by a single stroke were 0.35 +/- 0.09 pl. Interestingly, the amount of liquid adhered to the tip did not significantly depend on the protein concentration, but viscosity and surface tension of the sample. Under these conditions, we successfully obtained mass spectra for each biological sample. PMID- 19306265 TI - What happens with ageing: decline or remodeling of opsonin-independent phagocyte oxidative activity? AB - Changes that occur with age in the opsonin-independent oxidative activity of peripheral phagocytes in whole blood were examined by means of luminol chemiluminescence (LCL). The chemiluminescence was registered simultaneously by non-stimulated and stimulated cells and the age-related alterations of total and extracellular generation of reactive oxygen species (ROS) were studied using model systems. It was found that the rate of phagocyte activation by the glass surface of the measuring chambers, assessed by the time of the peak appearance after the start of LCL response, increased. However, the maximum oxidative activity and the integral oxidative capacity of the cells during adhesion, evaluated by the maximum LCL intensity and the area under the LCL curve, respectively, declined. No age-dependence of formyl-methionyl-leucyl phenylalanine (fMLP)-stimulated oxidative cellular activity for total ROS generation was detected. The maximum oxidative activity and the integral oxidative capacity of peripheral phagocytes to generate extracellular superoxide in response to fMLP was decreased. The likely causes for the observed alterations in phagocyte function are discussed and an analysis of the obtained results is given on the background of the contradictory data published on phagocyte oxidative activity age-related changes. PMID- 19306266 TI - Isolation of rare cells from cell mixtures by dielectrophoresis. AB - The application of dielectrophoretic field-flow fractionation (depFFF) to the isolation of circulating tumor cells (CTCs) from clinical blood specimens was studied using simulated cell mixtures of three different cultured tumor cell types with peripheral blood. The depFFF method can not only exploit intrinsic tumor cell properties so that labeling is unnecessary but can also deliver unmodified, viable tumor cells for culture and/or all types of molecular analysis. We investigated tumor cell recovery efficiency as a function of cell loading for a 25 mm wide x 300 mm long depFFF chamber. More than 90% of tumor cells were recovered for small samples but a larger chamber will be required if similarly high recovery efficiencies are to be realized for 10 mL blood specimens used CTC analysis in clinics. We show that the factor limiting isolation efficiency is cell-cell dielectric interactions and that isolation protocols should be completed within approximately 15 min in order to avoid changes in cell dielectric properties associated with ion leakage. PMID- 19306267 TI - Hexokinase inhibitor screening based on adenosine 5'-diphosphate determination by electrophoretically mediated microanalysis. AB - A CE-based method for hexokinase inhibitor screening was developed in the present paper. In this method, hexokinase activity was assayed via electrophoretically mediated microanalysis (EMMA), which combines on-column hexokinase-mediated reaction and measurement of produced adenosine 5'-diphosphate (ADP) via electrophoretical separation and UV detection. Enzyme inhibition can be read out directly from the reduced peak area of ADP in comparison with a reference electropherogram obtained in the absence of any inhibitor. Conditions for on column enzyme reaction and separation of adenosine 5'-triphosphate (ATP) and ADP were optimized. The optimal buffer composition for enzymatic reaction was 25 mM HEPES buffer (pH 7.5) containing 5 mM MgCl(2), whereas the optimal buffer composition for separation was 100 mM Tris-phosphate buffer (pH 5.5) containing 0.02% (m/v) hexadimethrine bromide (HDB). Fortunately, discontinuous buffer system can be adapted easily in the EMMA method. The time for separation was reduced dramatically to less than 3 min by reversing the direction of EOF via dynamically coating the capillary wall with the cationic polyelectrolyte HDB. Moreover, the peak tailing of ATP was also reduced by HDB coating. The Z' factor as high as 0.98 was obtained, indicating a high quality of the screening data. The present method is simple, robust and cost-effective. PMID- 19306268 TI - Accurate quantification of DNA methylation of DRD4 applying capillary gel electrophoresis with LIF detection. AB - Aberrant DNA methylation of gene promoters may be investigated by an array of different technologies. Besides DNA sequencing techniques following bisulfite treatment and determination of overall methylation by quantification of 5 methylcytosine/cytosine ratio following DNA hydrolysis, most approaches rely on PCR amplification of a defined template and subsequent analysis by conventional gel electrophoresis. As an additional analytical tool, a capillary gel electrophoresis method has been developed to quantify the methylation in combined bisulfite restriction analysis products of the gene dopamine receptor D4 (DRD4). Analyses were carried out in a bare fused-silica capillary dynamically coated with a 1% w/w solution of PVA (M(r)=72,000). A buffer (pH 7.3) containing 3% w/w 2-hydroxyethylcellulose (M(nu) approximately 90,000 g/mol) was used as sieving matrix. With 1/x weighted regression the accuracy (bias) of the method is within +/-10% and the precision (expressed as RSD) also meets the common acceptance criteria of 15% (20% near lower LOQ). It overcomes the limitations of standard gel electrophoresis, which allows only one single run per analysis and requires large amounts of DNA. Therefore, the method represents a valuable tool for routine quantitative analysis of the methylation status of DRD4 and other target genes. PMID- 19306269 TI - CE can identify small molecules that selectively target soluble oligomers of amyloid beta protein and display antifibrillogenic activity. AB - Soluble and toxic oligomers of amyloid beta (A beta) protein have been identified as the true neurotoxic species involved in Alzheimer's disease and considering them as targets to inhibit A beta aggregation might have a therapeutic value. We previously set up a CE method that enables the separation and quantification of transient oligomers of A beta protein-containing 42 amino acids (A beta(1-42)) along the pathway leading to fibrils and we now demonstrate how this method can be successfully applied to examine the in vitro inhibitory effects of small molecules on A beta oligomerization. To this end, we investigated mitoxantrone and pixantrone, two well-known anticancer drugs, as well as suramin and a suramin like compound. By using CE, it is here shown how mitoxantrone and pixantrone either reduce or block A beta(1-42) oligomerization, while Thioflavin T spectrofluorimetric assay and transmission electron microscopy demonstrate how these two compounds also display antifibrillogenic activity. Interestingly, in vitro cell viability experiments indicated that pixantrone significantly reduces A beta(1-42) neurotoxicity. PMID- 19306270 TI - Angioarchitecture of the venous and capillary system in heart defects induced by retinoic acid in mice. AB - BACKGROUND: Corrosion casting and immunohistochemical staining with anti-alpha smooth muscle actin and anti-CD34 was utilized to demonstrate the capillary plexus and venous system in control and malformed mouse hearts. METHODS: Outflow tract malformations (e.g., double outlet right ventricle, transposition of the great arteries, and common truncus arteriosus) were induced in progeny of pregnant mice by retinoic acid administration at day 8.5 of pregnancy. RESULTS: Although control hearts exhibited areas in which capillaries tended to be oriented in parallel arrays, the orientation of capillaries in the respective areas of malformed hearts was chaotic and disorganized. The major branch of a conal vein in control hearts runs usually from the left side of the conus to its right side at the root of the pulmonary trunk and opens to the right atrium below the right auricle; thus, it has a curved course. On the other hand, a conal vein in malformed hearts courses from the left side or from the anterior side of the conus and tends to traverse straight upwards along the dextroposed aorta or along the aortopulmonary groove with its proximal part located outside of the heart. Other cardiac veins in outflow tract malformations are positioned in the same locations as in control hearts. CONCLUSIONS: We postulate that the changed location of the conal vein and disorganized capillary plexus result from malformed morphogenesis of the outflow tract and/or a disturbed regulation of angiogenic growth factor release from the adjacent environment. PMID- 19306271 TI - Introduction: Links between social interaction and executive function. AB - The term executive function is used increasingly within developmental psychology and is often taken to refer to unfolding brain processes. We trace the origins of research on executive function to show that the link with social interaction has a long history. We suggest that a recent frenzy of research exploring methods for studying individual executive skills should pay more attention to the tradition exploring the role of social interaction in their development. PMID- 19306272 TI - Parental scaffolding and the development of executive function. AB - Research has demonstrated that differential parental scaffolding utterances influence children's development of executive function. Traditional conceptualizations of scaffolding, though, have difficulty in explaining how such differential effects influence children's cognitive development; they do not account for the timing of parental utterances with respect to children's currently occurring activities. We present a study examining the relationship between the timing of different parental scaffolding utterances and children's attention-switching EF abilities. There was a strong relation between the timing of elaborative parental utterances and attention switching. We discuss the implications of the findings for the conceptualization of the scaffolding process. PMID- 19306273 TI - How do families help or hinder the emergence of early executive function? AB - This chapter describes longitudinal findings from a socially diverse sample of 125 British children seen at ages two and four. Four models of social influence on executive function are tested, using multiple measures of family life as well as comprehensive assessments of children's executive functions. Our results confirm the importance of maternal scaffolding for young children's executive functions, but they also suggest positive effects of observational learning and adverse effects of disorganized and unpredictable family life; however, no support was found for an association between executive function and general positive characteristics of family interactions. PMID- 19306274 TI - New directions in evaluating social problem solving in childhood: early precursors and links to adolescent social competence. AB - A major objective of this chapter is to present a novel, ecologically sensitive social problem-solving task for school-aged children that captures the complexity of social and cognitive demands placed on children in naturalistic situations. Competence on this task correlates with a range of skills including executive functions, verbal reasoning, and attention. Children able to successfully carry out this task in middle school were more competent in early adolescence in collaborating in joint problem-solving tasks with peers and solving conflicts with parents. PMID- 19306275 TI - Culture, executive function, and social understanding. AB - Much of the evidence from the West has shown links between children's developing self-control (executive function), their social experiences, and their social understanding (Carpendale & Lewis, 2006, chapters 5 and 6), across a range of cultures including China. This chapter describes four studies conducted in three Oriental cultures, suggesting that the relationships among social interaction, executive function, and social understanding are different in these cultures, implying that social and executive skills are underpinned by key cultural processes. PMID- 19306276 TI - Social origins of executive function development. AB - The chapters in this issue revisit the social origins of the development of executive function (EF) through both empirical examination of the contexts in which EF development occurs (in vivo), as well as its social antecedents and consequences. Importantly, they also point to new directions in studying the social foundations of neurodevelopment, novel methods that take the social context into account, and cultural influences on EF development. PMID- 19306277 TI - Checkpoint responses to unusual structures formed by DNA repeats. AB - DNA sequences that are prone to adopting non-B DNA secondary structures are associated with hotspots of genomic instability. The fine mechanisms by which alternative DNA structures induce phenomena such as repeat expansions, chromosomal fragility, or gross chromosomal rearrangements are under intensive studies. It is well established that DNA damage checkpoint responses play a crucial role in maintaining a stable genome. It is far less clear, however, whether and how the checkpoint machinery responds to alternative DNA structures. This review discusses the role of the interplay between DNA damage checkpoints and alternative DNA structures in genome maintenance. PMID- 19306278 TI - Monozygotic female twins discordant for phenotype of Wilson's disease. AB - Wilson's disease (WD) is an autosomal recessive disorder characterized by the functional disruption of the copper-transporting protein adenosine triphosphatase 7B (ATP-ase 7B). The disease is caused by mutations in ATP7B gene. It seems that the type of mutation in ATP7B only to some degree determines phenotypic manifestation of WD. We examined two pairs of monozygotic twins discordant for WD phenotype. The first set of twins were ATP7B compound heterozygotes c.3207C>A (p.H1069Q)/c.1211_1212insA (p.N404Kfs). The index case developed severe liver failure followed by depressive symptoms, dysarthria, and tremor at the age of 36. Her sister remained presymptomatic at diagnosis at the age of 39. The second twins were ATP7B c.3207C.A (p.H1069Q) homozygotes. The index case presented with dysarthria and tremor at the age of 26. Her sister remained clinically presymptomatic at diagnosis at the age of 28. We concluded that the phenotypic characteristics of WD are possibly attributable to epigenetic/environmental factors. PMID- 19306279 TI - Determinants of quality of life in children with Gilles de la Tourette syndrome. AB - The objective of this study is to assess the association between tic severity, attention deficit disorder, obsessive-compulsive behavior, and quality of life (QOL) in children with Gilles de la Tourette syndrome (GTS). GTS is a multidimensional disorder with disturbances in motor function and behavior. However, little is known about what variables are associated with QOL in these patients. We evaluated 56 outpatients with a diagnosis of GTS. The mean age was 10 (range 5-17 years). Tics were assessed with the Yale Global Tic Severity Scale (YGTSS). Behavioral scales included the Leyton Obsessional Inventory-Child Version, Children's Yale-Brown Obsessive Compulsive Scale (CY-BOCS), and Attention-Deficit/Hyperactivity Disorder (ADHD) rating scale. The patient's parent also completed the TNO-AZL Children's Quality of Life scale (TACQOL). YGTSS scores ranged from 4 to 30, indicating mild to moderate tic severity. Motor and phonic tic ratings were not correlated with QOL. However, both ADHD and OCD were significantly related to QOL. Subanalysis of ADHD subtypes demonstrated that inattentiveness but not hyperactivity predicted lower QOL. When ADHD, Leyton OCD, and tic severity were considered simultaneously, tic severity remained non significant, while both ADHD and OCD remained significant contributors to QOL. In summary, in patients with mild to moderate GTS, QOL relates primarily to co morbidities of ADHD and obsessive-compulsive behavior. ADHD with predominantly inattentive symptoms, rather than hyperactivity symptoms, was associated with lower QOL. To improve QOL, clinicians must consider treatments of co-morbidities among tic patients. PMID- 19306280 TI - Sequence context-specific mutagenesis and base excision repair. AB - Base excision repair (BER) is critical for the maintenance of genome stability because it repairs at least 20,000 endogenously generated DNA lesions/cell/d. Several enzymes within the BER pathway exhibit sequence context dependency during the excision and DNA synthesis steps of repair. New evidence is emerging that germ line and tumor-associated variants of enzymes in this repair pathway exhibit sequence context dependence that is different from their ancestral counterparts. We review what is known about the ancestral and variant BER proteins within various sequence contexts. We suggest that altering the sequence context preferences of BER proteins could give rise to rare cellular variants that might have a selective advantage in response to environmental exposure or to the tumor microenvironment. PMID- 19306281 TI - The use of tunable diode laser absorption spectroscopy for rapid measurements of the delta13C of animal breath for physiological and ecological studies. AB - In this study we introduce the use of tunable diode laser absorption spectroscopy (TDLAS) as a technique for making measurements of the delta13C of animal 'breath' in near real time. The carbon isotope ratios (delta13C) of breath CO2 trace the carbon source of the materials being metabolized, which can provide insight into the use of specific food resources, e.g. those derived from plants using C3 versus C4 or CAM photosynthetic pathways. For physiological studies, labeled substrates and breath analyses provide direct evidence of specific physiological (e.g. fermentative digestion) or enzymatic (e.g. sucrase activity) processes. Although potentially very informative, this approach has rarely been taken in animal physiological or ecological research. In this study we quantify the utilization of different plant resources (photosynthetic types--C3 or C4) in arthropod herbivores by measuring the delta13C of their 'breath' and comparing it with bulk tissue values. We show that breath delta13C values are highly correlated with bulk tissues and for insect herbivores reflect their dietary guild, in our case C3-specialists, C4-specialists, or generalists. TDLAS has a number of advantages that will make it an important tool for physiologists, ecologists and behaviorists: it is non-invasive, fast, very sensitive, accurate, works on animals of a wide range of body sizes, per-sample costs are small, and it is potentially field-deployable. PMID- 19306282 TI - Fragmentation reactions of some peptide b3 ions: an energy-resolved study. AB - The fragmentation reactions of b3 ions of nominal structure AAAoxa, YAAoxa, AYAoxa and AAYoxa have been studied as a function of collision energy, allowing the construction of breakdown graphs expressing in a qualitative way the energy dependence of the fragmentation reactions. The primary fragmentation reactions of the AAAoxa b3 ion involve formation of the a3* (a3-NH3) ion and the b2 ion, with the latter becoming the dominant product at higher internal energies. For both YAAoxa and AYAoxa b3 ions the pathway to a3* is relatively minor with formation of b2 the dominant primary fragmentation reaction. For the AAYoxa b3 ion, in addition to a3*, abundant formation of the tyrosine (Y) iminium ion is observed with only minor formation of the b2 ion. The results support and expand upon the detailed mechanism of fragmentation of b3 ions proposed by Cooper et al. (J. Am. Soc. Mass Spectrom. 2006; 17: 1654). PMID- 19306283 TI - Evaluating uncertainty in the calculation of non-exchangeable hydrogen fractions within organic materials. AB - We calculated the fraction of exchangeable hydrogen atoms in proteinaceous materials commonly analyzed for stable isotopic composition related to the region of-origin of an animal. These included several types of alpha- and beta-keratin, and muscle tissue. We find that the fraction of H atoms in keratin available for exchange at a biologically relevant temperature (25 degrees C) averaged 9% across a range of ground organic materials, but was as high as approximately 17% in cut hair; muscle tissue has approximately 12% exchangeable H atoms. Under most analysis conditions, the difference in exchangeable fractions due to physical sample processing has a minimal effect on the calculated delta2H values of the non-exchangeable H atoms within a keratin-containing tissue (<2 per thousand). However, extreme mismatches between sample and reference material types could affect delta2H values. PMID- 19306284 TI - A thermal decomposition study of polymers by tunable synchrotron vacuum ultraviolet photoionization mass spectrometry. AB - The thermal decomposition of polymers (poly(vinyl chloride) (PVC) and polystyrene (PS)) has been studied with synchrotron VUV photoionization mass spectrometry at low pressure. Pyrolysis products formed at different temperatures have been identified by the measurement of photoionization mass spectra at different photon energies. The experimental results demonstrate the variation of the pyrolysis product pool of PVC at different temperatures, dividing the thermal decomposition process into two stages: the low-temperature stage to form HCl and benzene, and the high-temperature stage to form numerous large aromatic hydrocarbons. For the thermal decomposition of PS, four reaction categories are determined. This work reports a new application of synchrotron VUV photoionization mass spectrometry in the study of the thermal decomposition of polymers, and demonstrates its good performance in product analysis, which is expected to help understand the thermal decomposition mechanism of PVC, PS and other synthesized polymers. PMID- 19306285 TI - Study of the oxidation dynamics of ethyl cysteinate dimer in solution by ultra performance liquid chromatography with tandem mass spectrometry. AB - Diaminodithiol (N2S2)-type compounds readily oxidize to produce disulfides. We found that some ligands failed to produce a prospective protonated molecular ion peak but gave a peak of [M-2+H]+, whereas others produced both [M+H]+ and [M 2+H]+ peaks in electrospray ionization mass spectra. In this study, an important N2S2 ligand, the ethyl cysteinate dimer (ECD), was investigated with high resolution accurate mass measurements and tandem mass spectrometric analysis. The elemental compositions of ECD and its oxidized product were analyzed. The oxidation of ECD was confirmed. An ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) method in multiple reaction monitoring mode was developed, and ECD and its oxidized product were quantitated in solution. The dynamic oxidation process of ECD in solution was studied in detail. The full time course of the decrease in ECD and the increase in its oxide was observed; the oxidation procedure followed first-order kinetics, and the half-life time of ECD was 51 min. PMID- 19306286 TI - Matrix/analyte ratio influencing polymer molecular weight distribution in matrix assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has been used to characterize poly(L-lysine) polymers and unique oligomer peptides, like 10-, 15- and 25-mer [Lys]n oligolysine peptides. Several matrices have been tried in order to find optimal conditions, but only alpha cyano-4-hydroxycinnamic acid gave analytically useful spectra. The synthetic oligomers and their mixtures gave good quality spectra, showing protonated and cationized molecules, including doubly charged species. The polymers, analogously, gave a wide distribution of single- and double-cationized peak series. The polymer distributions observed indicate the presence of significant suppression effects. The concentration (matrix/analyte ratio) was found to influence the results significantly; distributions shifting to higher masses when higher polymer concentrations were used. This effect was studied in detail using the synthetic ('monodisperse') oligolysine peptides. It was found that the relative intensities change by over an order of magnitude in the 0.1-10 pmol/microL concentration range (typical for most proteomic analyses). The results indicate that concentration effects need to be considered when MALDI-MS is used for quantitative purposes. PMID- 19306287 TI - Study of bisphosphonates by matrix-assisted laser desorption/ionization mass spectrometry--influence of alkali atoms on fragmentation patterns. AB - 1-hydroxymethylene-1,1-bisphosphonic acids (or bisphosphonates) are compounds that have interesting pharmacological applications. However, few mass spectrometric investigations have been carried out to determine their fragmentation patterns. Herein, we evaluated different matrices for the study by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) of the formation and fragmentation of the protonated, the cationized (MNa+ and MK+) and the deprotonated bisphosphonates. Some in-source fragmentations were observed both in positive and in negative ion modes. The fragmentation patterns obtained in post-source decay mode are also discussed. In contrast to previous electrospray ionization/multi-stage mass spectrometry (ESI MSn) studies, some new fragmentation pathways were deduced and the effects of alkali ions on the fragmentation patterns were shown. The results summarized here completed the data previously recorded by ESI-MSn and could be used for the characterization of bisphosphonates as alkali complexes in biological mixtures. PMID- 19306288 TI - Efficient encoding and rapid decoding for interactive visualization of large three-dimensional hyperspectral chemical images. AB - Interactive visualization of data from a new generation of chemical imaging systems requires coding that is efficient and accessible. New technologies for secondary ion mass spectrometry (SIMS) generate large three-dimensional, hyperspectral datasets with high spatial and spectral resolution. Interactive visualization is important for chemical analysis, but the raw dataset size exceeds the memory capacities of typical current computer systems and is a significant obstacle. This paper reports the development of a lossless coding method that is memory efficient, enabling large SIMS datasets to be held in fast memory, and supports quick access for interactive visualization. The approach provides pixel indexing, as required for chemical imaging applications, and is based on the statistical characteristics of the data. The method uses differential time-of-flight to effect mass-spectral run-length-encoding and uses a scheme for variable-length, byte-unit representations for both mass-spectral time-of-flight and intensity values. Experiments demonstrate high compression rates and fast access. PMID- 19306289 TI - Sensory functions in dystonia: insights from behavioral studies. AB - The pathophysiology of primary dystonia is thought to involve dysfunction of the basal ganglia cortico-striatal-thalamo-cortical motor circuits. In the past, emphasis was placed on the role of the basal ganglia in controlling movements; in more recent times, however, it has also become clear that they play an important part in sensory as well as cognitive functions. Here, we review evidence for dysfunction of sensory processing in patients with dystonia, and speculate that this may lead to abnormalities in a crucial role of the basal ganglia that links sensory information to appropriate motor output. Sensory function, particularly in the somatosensory domain, has been shown to be compromised in patients with primary dystonia, both in adult onset focal dystonia and in genetically characterized DYT1 dystonia. Given that nonaffected DYT1 gene carriers may show similar abnormalities to clinically affected individuals, sensory deficits could constitute a subclinical endophenotypic trait of disease that precedes overt clinical manifestations. Whether they can trigger primary dystonia or are an epiphenomenon is an issue warranting further study, but the fact that a number of different neurorehabilitative approaches explicitly manipulate somatosensory inputs to improve motor function suggests there may be a causal link between them. We believe that in future, randomized, blind and controlled studies in large patient populations should address this issue, providing efficient strategies to aid functional recovery, particularly in focal hand dystonia, where the available medical treatments offer little benefit. PMID- 19306290 TI - Karl-Axel Ekbom and iron deficiency in restless legs syndrome. PMID- 19306291 TI - Pramipexole induced compulsive behaviors abate after initiation of rotigitine. PMID- 19306292 TI - Every microsatellite is different: Intrinsic DNA features dictate mutagenesis of common microsatellites present in the human genome. AB - Microsatellite sequences are ubiquitous in the human genome and are important regulators of genome function. Here, we examine the mutational mechanisms governing the stability of highly abundant mono-, di-, and tetranucleotide microsatellites. Microsatellite mutation rate estimates from pedigree analyses and experimental models range from a low of approximately 10(-6) to a high of approximately 10(-2) mutations per locus per generation. The vast majority of observed mutational variation can be attributed to features intrinsic to the allele itself, including motif size, length, and sequence composition. A greater than linear relationship between motif length and mutagenesis has been observed in several model systems. Motif sequence differences contribute up to 10-fold to the variation observed in human cell mutation rates. The major mechanism of microsatellite mutagenesis is strand slippage during DNA synthesis. DNA polymerases produce errors within microsatellites at a frequency that is 10- to 100-fold higher than the frequency of frameshifts in coding sequences. Motif sequence significantly affects both polymerase error rate and specificity, resulting in strand biases within complementary microsatellites. Importantly, polymerase errors within microsatellites include base substitutions, deletions, and complex mutations, all of which produced interrupted alleles from pure microsatellites. Postreplication mismatch repair efficiency is affected by microsatellite motif size and sequence, also contributing to the observed variation in microsatellite mutagenesis. Inhibition of DNA synthesis within common microsatellites is highly sequence-dependent, and is positively correlated with the production of errors. DNA secondary structure within common microsatellites can account for some DNA polymerase pause sites, and may be an important factor influencing mutational specificity. PMID- 19306293 TI - Differential role of beta-catenin in VEGF and histamine-induced MMP-2 production in microvascular endothelial cells. AB - Increases in endothelial cell permeability and production of matrix-degrading enzymes are two early steps in the angiogenic process. Factors such as vascular endothelial growth factor (VEGF) and histamine induce the angiogenic process through alterations in both permeability and proteolysis. We hypothesized that beta-catenin acts as a positive regulator of MMP-2 and MT1-MMP transcription following VEGF or histamine stimulation. Rat microvascular endothelial cells were exposed to VEGF or histamine overnight and MMP-2 protein production was assessed by gelatin zymography. Latent MMP-2 protein levels were increased following VEGF and histamine treatment as were MMP-2 mRNA transcript levels. Endothelial cells exposed to VEGF and histamine had an increased level of nuclear beta-catenin, which was sensitive to inhibition of the PI3-kinase signaling pathway. Promoter assays indicated increased transcriptional activity of both MMP-2 and MT1-MMP in endothelial cells co-transfected with luciferase reporter constructs and beta catenin. Inhibition of beta-catenin signaling with inhibitor of catenin and T cell factor (ICAT) revealed that the VEGF-induced increase in MMP-2 mRNA is beta catenin dependent. Interestingly, while MMP-2 mRNA levels increased in response to histamine H1 or H2 receptor activation, significantly larger increases were observed in cells co-treated with ICAT and histamine or the histamine receptor agonists, HTMT and dimaprit. While both VEGF and histamine increase nuclear beta catenin and MMP-2 production, the role of beta-catenin in MMP-2 regulation differs between the two stimuli. PMID- 19306294 TI - Sp proteins and Runx2 mediate regulation of matrix gla protein (MGP) expression by parathyroid hormone. AB - As part of its catabolic action in bone, parathyroid hormone (PTH) inhibits extracellular matrix mineralization. We previously showed that PTH dose dependently induces matrix gla protein (MGP) expression in osteoblasts and this induction is at least partially responsible for PTH-mediated inhibition of mineralization. Recently, we identified PKA and ERK/MAPK as the key signaling pathways involved in PTH regulation of MGP expression. The goal of this study was to further characterize the mechanism by which PTH stimulates expression of MGP. Deletion analysis of the murine Mgp gene promoter identified a PTH-responsive region between -173 bp and-49 bp. Using gel-mobility shift assays we found that Sp1/Sp3, and Runx2 bind to distinct sites within this region. Mutation of either the Sp or the Runx2 site reduced MGP induction by PTH, while mutation of both sites completely abolished PTH responsiveness. Overexpression of Runx2 or Sp1 activated the Mgp reporter, while Sp3 was a dose-dependent repressor of Sp1 and PTH-induced MGP expression. Collectively, these data show that PTH regulates MGP gene transcription in osteoblasts through altered activities of Sp and Runx2 transcription factors. PMID- 19306295 TI - Clinorotation upregulates inducible nitric oxide synthase by inhibiting AP-1 activation in human umbilical vein endothelial cells. AB - Alterations of nitric oxide contribute to post-flight orthostatic intolerance. The aim of this study was to investigate the changes of inducible nitric oxide synthase (iNOS) and the mechanisms underlying regulation of iNOS by simulated microgravity in human umbilical vein endothelial cells (HUVECs). Clinorotation, a simulated-model of microgravity, increased iNOS expression and promoter activity in HUVECs. The transactivations of NF-kappaB and AP-1 were suppressed by 24 h clinorotation. A key role for AP-1, but not NF-kappaB in the regulation of iNOS was shown. (1) PDTC, a NF-kappaB inhibitor, had no effect on clinorotation upregulation of iNOS. (2) SP600125, a JNK-specific inhibitor, which resulted in inhibition of AP-1 activity, enhanced the iNOS expression and promoter activity in clinorotation. (3) Overexpression of AP-1 remarkably attenuated the upregulation effect of clinorotation. These findings indicate that clinorotation upregulates iNOS in HUVECs by a mechanism dependent on suppression of AP-1, but not NF-kappaB. These results support a key role for AP-1 in the signaling of postflight orthostatic intolerance. PMID- 19306296 TI - Suppression of PAI-1 expression through inhibition of the EGFR-mediated signaling cascade in rat kidney fibroblast by ascofuranone. AB - Fibrosis in glomerulosclerosis causes progressive loss of renal function. Transforming growth factor (TGF)-beta, one of the major profibrotic cytokines, induces the synthesis of plasminogen activator inhibitor (PAI)-1, a factor that plays a crucial role in the development of fibrosis. Here, we found that an isoprenoid antibiotic, ascofuranone, suppresses expression of profibrotic factors including matrix proteins and PAI-1 induced by TGF-beta in renal fibroblasts. Ascofuranone selectively inhibits phosphorylation of epidermal growth factor receptor (EGFR), and downstream kinases such as Raf-1, MEK-1/2, and ERK-1/2. PAI 1 transcription also is suppressed by treatment with kinase inhibitors for MEK 1/2 or EGFR, and with small interfering RNA for EGFR. Ascofuranone inhibits cellular metalloproteinase activity, and an inhibitor of metalloproteinases suppresses EGFR phosphorylation and PAI-1 transcription. These results suggest that ascofuranone suppresses expression of profibrotic factors through the inhibition of an EGFR-dependent signal transduction pathway activated by metalloproteinases. PMID- 19306297 TI - Development of the transgenic cyan fluorescent protein (CFP)-expressing nude mouse for "Technicolor" cancer imaging. AB - A major goal for in vivo biology is to develop models which can express multiple colors of fluorescent proteins in order to image many processes simultaneously in real time. Towards this goal, the cyan fluorescent protein (CFP) nude mouse was developed by crossing non-transgenic nude mice with the transgenic CK/ECFP mouse in which the beta-actin promoter drives expression of CFP in almost all tissues. In crosses between nu/nu CFP male mice and nu/+ CFP female mice, approximately 50% of the embryos fluoresced blue. In the CFP nude mice, the pancreas and reproductive organs displayed the strongest fluorescent signals of all internal organs which vary in intensity. Orthotopic implantation of XPA-1 human pancreatic cancer cells expressing red fluorescent protein (RFP); or green fluorescent protein (GFP) in the nucleus and RFP in the cytoplasm, was performed in female nude CFP mice. Color-coded fluorescence imaging of these human pancreatic cancer cells implanted into the bright blue fluorescent pancreas of the CFP nude mouse afforded novel insight into the interaction of the pancreatic tumor and the normal pancreas, in particular the strong desmoplastic reaction of the tumor. The naturally enhanced blue fluorescence of the pancreas in the CFP mouse serves as an ideal background for color-coded imaging of the interaction of implanted cancer cells and the host. The CFP nude mouse will provide unique understanding of the critical interplay between the cancer cells and their microenvironment. PMID- 19306298 TI - c-Abl modulates AICD dependent cellular responses: transcriptional induction and apoptosis. AB - APP intracellular domain (AICD) has been proposed as a transcriptional inductor that moves to the nucleus with the adaptor protein Fe65 and regulates transcription. The two proteins, APP and Fe65, can be phosphorylated by c-Abl kinase. Neprilysin has been proposed as a target gene for AICD. We found that AICD expression is decreased by treatment with STI-571, a c-Abl inhibitor, suggesting a modulation of AICD transcription by c-Abl kinase. We observed interaction between c-Abl kinase, the AICD fragment and the Fe65 adaptor protein. In addition, STI-571 reduces apoptosis in APPSw, and the apoptotic response induced by Fe65 over-expression was inhibited by with the expression of a kinase dead (KD) c-Abl and enhanced by over-expression of WT-c-Abl. However, in the APPSw cells, the ability of the KD-c-Abl to protect against Fe65 was reduced. Finally, in APPSw clone, we detected higher trans-activation of the pro-apoptotic p73 isoform, TAp73 promoter. Our results show that c-Abl modulates AICD dependent cellular responses, transcriptional induction as well as the apoptotic response, which could participate in the onset and progression of the neurodegenerative pathology, observed in Alzheimer's disease (AD). PMID- 19306299 TI - Cocaine adulterants used as marker compounds. PMID- 19306300 TI - Let eating disorder patients decide: Providing choice may reduce early drop-out from inpatient treatment. AB - Premature drop-out from treatment is a highly prevalent phenomenon among eating disorder (ED) patients. In a specialized inpatient treatment unit a major change was made in the admission strategy in 2001, giving a maximum of personal choice to the patients. A quasi-experimental research was carried out comparing 87 patients treated till 2000 ('old' strategy) with 87 patients treated from 2001 on ('new' strategy). The results indicate that the provision of choice at the beginning of treatment significantly reduced drop-out during the first weeks of inpatient treatment. No differences between both strategies on later drop-out and weight change (in anorexia nervosa patients) during inpatient treatment were found. The results are discussed in the light of the importance placed on dynamics of personal choice, autonomy and volition within the framework of the self-determination theory (SDT). PMID- 19306301 TI - Individual blood-cell capture and 2D organization on microarrays. PMID- 19306302 TI - Ultrasensitive electrochemical DNA biosensors based on the detection of a highly characteristic solid-state process. PMID- 19306303 TI - Psychological resilience and postdeployment social support protect against traumatic stress and depressive symptoms in soldiers returning from Operations Enduring Freedom and Iraqi Freedom. AB - BACKGROUND: A number of studies have examined the prevalence and correlates of posttraumatic stress disorder (PTSD), depression, and related psychiatric conditions in soldiers returning from Operations Enduring Freedom and Iraqi Freedom (OEF/OIF), but none have examined whether factors such as psychological resilience and social support may protect against these conditions in this population. METHODS: A total of 272 predominantly older reserve/National Guard OEF/OIF veterans completed a mail survey assessing traumatic stress and depressive symptoms, resilience, and social support. RESULTS: Resilience scores in the full sample were comparable to those observed in civilian outpatient primary-care patients. Respondents with PTSD, however, scored significantly lower on this measure and on measures of unit support and postdeployment social support. A hierarchical regression analysis in the full sample suggested that resilience (specifically, increased personal control and positive acceptance of change) and postdeployment social support were negatively associated with traumatic stress and depressive symptoms, even after adjusting for demographic characteristics and combat exposure. CONCLUSIONS: These results suggest that interventions to bolster psychological resilience and postdeployment social support may help reduce the severity of traumatic stress and depressive symptoms in OEF/OIF veterans. PMID- 19306304 TI - Diurnal mood variation in outpatients with major depressive disorder. AB - BACKGROUND: Diurnal mood variation (DMV) with early morning worsening is considered a classic symptom of melancholic features of major depressive disorder (MDD) according to the Diagnostic and Statistical Manual. This report used data from the sequenced treatment alternatives to relieve depression study to determine whether DMV was associated with treatment outcome to citalopram. METHODS: Two thousand eight hundred and seventy-five outpatients with nonpsychotic MDD were evaluated during a 14-week trial of the selective serotonin reuptake inhibitor citalopram. Participants were divided into three groups: those with "classic" DMV (early morning worsening), those with any form of DMV (morning, afternoon, or evening worsening), and those with no DMV. Participants with classic DMV and those with any form of DMV were compared to those with no DMV in terms of baseline sociodemographic and clinical characteristics, treatment outcomes, and treatment features. RESULTS: Minor baseline clinical characteristics and treatment feature differences were found between participants with and without DMV. Participants with classic morning DMV had slightly higher response rates than those without DMV. However, no differences were found in response or remission between either group of participants with DMV and those with no DMV. CONCLUSION: DMV does not appear to be associated with a unique prominent pattern of response to selective serotonin reuptake inhibitor treatment in patients with depression, and does not appear to be a serotonergically modulated process. Further evaluation is necessary to determine if this relationship holds true for dopaminergic and noradrenergic antidepressant agents, such as dual-acting agents or antidepressant medication combinations. PMID- 19306305 TI - Antidepressant use in a nationally representative sample of community-dwelling US Latinos with and without depressive and anxiety disorders. AB - BACKGROUND: Antidepressant drugs are among the most widely prescribed drugs in the United States; however, little is known about their use among major ethnic minority groups. METHOD: Collaborative Psychiatric Epidemiology Surveys (CPES) data were analyzed to calculate nationally representative estimates of Latino and non-Latino White adults antidepressant use. SETTING: The 48 coterminous United States was the setting. PARTICIPANTS: Household residents aged 18 years and older (N=9,250). MAIN OUTCOME: Past year antidepressant use. RESULTS: Compared to non Latino Whites, few Latinos, primarily Mexican Americans, with 12-month depressive and/or anxiety disorders reported past year antidepressant use. Mexican Americans (OR=0.48; 95%CI=0.30-0.77) had significantly lower odds of use compared to non Latino Whites, which were largely unaffected by factors associated with access to care. Over half of antidepressant use was by respondents not meeting 12-month criteria for depressive or anxiety disorders. Lifetime depressive and anxiety disorders explained another 21% of past year antidepressant use, leaving another 31% of drug use unexplained. DISCUSSION: We found a disparity in antidepressant use for Mexican Americans compared to non-Latino Whites that was not accounted for by differences in need and factors associated with access to care. About one third of antidepressant use was by respondents not meeting criteria for depressive or anxiety disorders. Our findings underscore the importance of disaggregating Latino ethnic groups. Additional work is needed to understand the medical and economic value of antidepressant use beyond their primary clinical targets. PMID- 19306306 TI - The guinea pig as an animal model for developmental and reproductive toxicology studies. AB - BACKGROUND: Regulatory guidelines for developmental and reproductive toxicology (DART) studies require selection of "relevant" animal models as determined by kinetic, pharmacological, and toxicological data. Traditionally, rats, mice, and rabbits are the preferred animal models for these studies. However, for test articles that are pharmacologically inactive in the traditional animal models, the guinea pig may be a viable option. This choice should not be made lightly, as guinea pigs have many disadvantages compared to the traditional species, including limited historical control data, variability in pregnancy rates, small and variable litter size, long gestation, relative maturity at birth, and difficulty in dosing and breeding. METHODS: This report describes methods for using guinea pigs in DART studies and provides results of positive and negative controls. Standard study designs and animal husbandry methods were modified to allow mating on the postpartum estrus in fertility studies and were used for producing cohorts of pregnant females for developmental studies. RESULTS: A positive control study with the pregnancy-disrupting agent mifepristone resulted in the anticipated failure of embryo implantation and supported the use of the guinea pig model. Control data for reproductive endpoints collected from 5 studies are presented. CONCLUSION: In cases where the traditional animal models are not relevant, the guinea pig can be used successfully for DART studies. PMID- 19306307 TI - Intrinsic genomic instability from naturally occurring DNA structures: An introduction to the special issue. PMID- 19306308 TI - Non-B DNA conformations as determinants of mutagenesis and human disease. AB - Repetitive DNA motifs may fold into non-B DNA structures, including cruciforms/hairpins, triplexes, slipped conformations, quadruplexes, and left handed Z-DNA, thereby representing chromosomal targets for DNA repair, recombination, and aberrant DNA synthesis leading to repeat expansion or genomic rearrangements associated with neurodegenerative and genomic disorders. Hairpins and quadruplexes also determined the relative abundances of simple sequence repeats (SSR) in vertebrate genomes, whereas strong base stacking has permitted the expansion of purine.pyrimidine-rich SSR during evolutionary time. SSR are enriched in regulatory and cancer-related gene classes, where they have been actively recruited to participate in both gene and protein functions. SSR polymorphic alleles in the population are associated with cancer susceptibility, including within genes that appear to share regulatory circuits involving reactive oxygen species. PMID- 19306309 TI - Peptide nucleic acid (PNA) binding and its effect on in vitro transcription in friedreich's ataxia triplet repeats. AB - Peptide nucleic acids (PNAs) are DNA mimics in which peptide-like linkages are substituted for the phosphodiester backbone. Homopyrimidine PNAs can invade double-stranded DNA containing the homologous sequence by displacing the homopyrimidine strand from the DNA duplex and forming a PNA/DNA/PNA triplex with the complementary homopurine strand. Among biologically interesting targets for triplex-forming PNA are (GAA/CTT)(n) repeats. Expansion of these repeats results in partial inhibition of transcription in the frataxin gene, causing Friedreich's ataxia. We have studied PNA binding and its effect on T7 RNA polymerase transcription in vitro for short repeats (n = 3) and for long repeats (n = 39), placed in both possible orientations relative to the T7 promoter such that either the GAA-strand, or the CTT-strand serves as the template for transcription. In all cases PNA bound specifically and efficiently to its target sequence. For the short insert, PNA binding to the template strand caused partial transcription blockage with well-defined sites of RNA product truncation in the region of the PNA-binding sequence, whereas binding to the nontemplate strand did not block transcription. However, PNA binding to long repeats, whether in the template or the nontemplate strand, resulted in a dramatic reduction of the amount of full length transcription product, although in the case of the nontemplate strand there were no predominant truncation sites. Biological implications of these results are discussed. PMID- 19306310 TI - Selection for the G4 DNA motif at the 5' end of human genes. AB - Formation of G4 DNA may occur in the course of replication and transcription, and contribute to genomic instability. We have quantitated abundance of G4 motifs and potential for G4 DNA formation of the nontemplate strand of 5' exons and introns of transcripts of human genes. We find that, for all human genes, G4 motifs are enriched in 5' regions of transcripts relative to downstream regions; and in 5' regulatory regions relative to coding regions. Notably, although tumor suppressor genes are depleted and proto-oncogenes enriched in G4 motifs, abundance of G4 motifs in the 5' regions of transcripts of genes in these categories does not differ. These results support the hypothesis that G4 motifs are under selection in the human genome. They further show that for tumor suppressor genes and proto oncogenes, independent selection determines potential for G4 DNA formation of 5' regulatory regions of transcripts and downstream coding regions. PMID- 19306311 TI - Genetic instabilities of (CCTG).(CAGG) and (ATTCT).(AGAAT) disease-associated repeats reveal multiple pathways for repeat deletion. AB - The DNA repeats (CTG).(CAG), (CGG).(CCG), (GAA).(TTC), (ATTCT).(AGAAT), and (CCTG).(CAGG), undergo expansion in humans leading to neurodegenerative disease. A genetic assay for repeat instability has revealed that the activities of RecA and RecB during replication restart are involved in a high rate of deletion of (CTG).(CAG) repeats in E. coli. This assay has been applied to (CCTG).(CAGG) repeats associated with myotonic dystrophy type 2 (DM2) that expand to 11 000 copies and to spinocerebellar ataxia type 10 (SCA10) (ATTCT).(AGAAT) repeats that expand to 4500 copies in affected individuals. DM2 (CCTG).(CAGG) repeats show a moderate rate of instability, less than that observed for the myotonic dystrophy type 1 (CTG).(CAG) repeats, while the SCA10 (ATTCT).(AGAAT) repeats were remarkably stable in E. coli. In contrast to (CTG).(CAG) repeats, deletions of the DM2 and SCA10 repeats were not dependent on RecA and RecB, suggesting that replication restart may not be a predominant mechanism by which these repeats undergo deletion. These results suggest that different molecular mechanisms, or pathways, are responsible for the instability of different disease-associated DNA repeats in E. coli. These pathways involve simple replication slippage and various sister strand exchange events leading to deletions or expansions, often associated with plasmid dimerization. The differences in the mechanisms of repeat deletion may result from the differential propensity of these repeats to form various DNA secondary structures and their differential proclivity for primer template misalignment during replication. PMID- 19306312 TI - Editorial: photodissociation. PMID- 19306313 TI - A new method for the comparison of survival distributions. AB - The assessment of overall homogeneity of time-to-event curves is a key element in survival analysis in biomedical research. The currently commonly used testing methods, e.g. log-rank test, Wilcoxon test, and Kolmogorov-Smirnov test, may have a significant loss of statistical testing power under certain circumstances. In this paper we propose a new testing method that is robust for the comparison of the overall homogeneity of survival curves based on the absolute difference of the area under the survival curves using normal approximation by Greenwood's formula. Monte Carlo simulations are conducted to investigate the performance of the new testing method compared against the log-rank, Wilcoxon, and Kolmogorov Smirnov tests under a variety of circumstances. The proposed new method has robust performance with greater power to detect the overall differences than the log-rank, Wilcoxon, and Kolmogorov-Smirnov tests in many scenarios in the simulations. Furthermore, the applicability of the new testing approach is illustrated in a real data example from a kidney dialysis trial. PMID- 19306314 TI - Laboratory and field cage studies on female-targeted attract-and-kill bait stations for Anastrepha suspensa (Diptera: Tephritidae). AB - BACKGROUND: Development of attract-and-kill bait stations for pest fruit flies has been limited by the water solubility of sugar needed as a feeding stimulant and by the volatility of chemical attractants. A wax-based matrix was developed that provides the longevity needed for field use and is biodegradable. RESULTS: Laboratory bioassays with the Caribbean fruit fly, Anastrepha suspensa (Loew), confirmed the efficacy of bait stations containing avermectin, methomyl, spinosad and phloxine B. Field cage studies demonstrated that significant mortality occurred with either 1% (w/v) spinosad or 1% (w/v) methomyl bait stations versus pesticide-free bait stations. Bait stations were exposed to environmental conditions by placing them in trees at the ARS station in Miami, Florida, between tests. There was no loss in efficacy, in spite of exposure to over 360 mm of rainfall over the 56 days of the study, indicating that the bait stations could provide population suppression for at least 1-2 months when used in subtropical environments. CONCLUSION: A long-lasting, female-targeted fruit fly bait station, such as the one developed herein, could provide a cost-effective option for fruit fly population suppression that would be an important tool in tephritid pest management and control. Additional studies are needed to demonstrate efficacy against wild fruit fly populations and determine deployment strategies. PMID- 19306315 TI - Ceratocricoid muscle: an embryological and anatomical study. AB - This study aimed to document the prevalence and morphology of the ceratocricoid muscle in a large sample of fetuses and adults and to explain its possible origin in a sample of embryos. Forty-five embryos, thirty-four fetuses, and ninety human larynges from adults with no known laryngeal pathology were studied. The muscle was observed in 23% of the fetal sample and in 14% of the adult sample. No significant differences were observed in the laterality in any of the groups. The ceratocricoid muscle is attached between the cricoid lamina and the inferior horn of the thyroid cartilage and also into the capsule of the cricothyroid joint. The muscle is innervated by several branches (between one and three) from the anterior division of the recurrent laryngeal nerve as it courses behind the cricothyroid joint. The ceratocricoid muscle develops from tissue within the mesenchymal bridge which connects the external and internal laryngeal sphincters or rings from embryonic stages 15-20. The close relationship of the ceratocricoid muscle to the recurrent laryngeal nerve could mean that it can exert pressure on this nerve. This may be a possible explanation for the causation of certain idiopathic recurrent laryngeal nerve palsies. PMID- 19306316 TI - The long thoracic nerve: Its origin, branches, and relationship to the middle scalene muscle. AB - Anatomical knowledge regarding the long thoracic nerve (LTN) is important during surgical procedures considering that dysfunction of this nerve results in clinical problems. The purpose of this study was to explore the anatomy of the LTN, its origin, configuration, branching pattern, and relationship to the middle scalene muscle (MSM). The course of the LTN was investigated in 12 embalmed cadavers (21 sides). We defined four different types for this nerve according to the origins of its roots. The most common formation of the LTN was the contribution of three branches that originated from the fifth, sixth, and seventh cervical ventral roots. C5 and C6 components or upper portion of the LTN roots lay primarily between the middle and posterior scalene muscles, sometimes passed through the MSM, and less frequently coursed over the MSM. C7 contributions to the LTN were always located anterior to the MSM. Contributions from C8 were also found over the MSM. The median number of branches arising directly from the cervical roots and branches arising from the main trunk of the nerve were 3 and 7, respectively. Along its course, the median number of branches to the serratus anterior was 10. PMID- 19306317 TI - Landmark-based software for anatomical measurements: a precision study. AB - The aim of this study was to develop a software program, called Landmarker, which would aid studies of complex anatomical morphometry by simplifying the manual identification of landmarks in 3D images. We also tested its precision on routine magnetic resonance imaging (MRI) scans. To understand human biological variation, there is a need to identify morphological characteristics from the exterior and the interior of human anatomy. MRI, as opposed to other radiographic methods (mainly based on X-ray techniques), supplies good soft tissue contrast, which allows for more complex assessments than what bony landmarks can provide. Because automation of this assessment is highly demanding, one of the primary goals for the new software was to enable more rapid identification of landmark sets in 3D image data. Repeat acquisition of head MRIs having a resolution of 0.94 x 0.94 x 1.20 mm3 were performed on 10 volunteers. Intra- and interoperator, as well as interacquisition variations of manual identification of exterior, craniofacial interior, and brain landmarks were studied. The average distances between landmarks were <1.8 mm, <2.3 mm, and <2.0 mm in the intra- and interoperator, and interacquisition evaluations, respectively. This study presents new software for time efficient identification of complex craniofacial landmarks in 3D MRI. To the best of our knowledge, no evaluation of software for rapid landmark-based analysis of complex anatomies from 3D MR data has yet been presented. This software may also be useful for studies in other anatomical regions and for other types of image data. PMID- 19306318 TI - Gross morphology of the vastus lateralis muscle: An anatomical review. AB - To understand the role of the vastus lateralis (VL) muscle in the pathogenesis of common knee disorders such as patellofemoral joint syndrome, knowledge of its anatomical structure is essential. The aim of this study was to review currently available information on the gross morphology of VL. A structured literature review was undertaken and 36 references, comprising 22 scientific papers and 14 anatomical textbooks, were included. Results of this literature review show that most of the included studies exhibited methodological limitations, and focused on different parameters of the VL muscle. Hence, reproducibility of these studies and comparison of results was difficult. This review also demonstrates a dearth of information on the muscle architecture, compartmentalization, nerve supply and fusion of VL, and that there has been no investigation of the muscle as a whole unit. Further research is required of the architecture and innervation of the VL muscle to better understand its function. PMID- 19306319 TI - Short branch of type IV dual left anterior descending coronary artery running as an aberrant obtuse marginal branch: a previously undescribed arrangement. PMID- 19306320 TI - The problem of direct lymph drainage of the bronchopulmonary segments into the mediastinal and hilar lymph nodes. AB - The phenomenon of skip metastases depends on the occurrence of direct metastases of non-small cell lung cancer into mediastinal lymph nodes without the involvement of hilar lymph nodes. The medical literature suggests that this prevalence ranges between 13 and 37.8% of cases. The goal of our study was to evaluate the prevalence of subpleural superficial lymphatic vessels carrying the lymph from the bronchopulmonary segments directly to the mediastinal nodes thus skipping the hilar nodes and determine whether there is a tendency towards differentiation in lymph drainage between males and females. During autopsy, 27 left and 27 right lungs were removed from 19 male and eight female cadavers. The lymphatic vessels were visualized at the mediastinal and interlobar surface of the lung by visual inspection. These vessels were then cannulated and injected with drawing ink. The first lymph node to become ink-colored via injection was dissected and histologically examined. The lymph flowed into hilar lymph nodes in 79.5% of the cases and into the mediastinal nodes in 20.5% of all the 83 vessels injected; of these 13.2% were from the right lung and in 7.2% from the left lung. The upper right lobe (5/14 vessels) and its anterior segment (3/14 vessels) were the most common source of the direct lymph drainage to the mediastinum. Vessels of 15.4% in female cadavers (4/26) and 22.8% vessels in male cadavers (13/57) traveled directly to mediastinal nodes skipping the hilar nodes. This difference was not statistically significant. PMID- 19306321 TI - Variations in the MRI appearance of the insertion of the tendon of subscapularis. AB - Standard anatomical textbooks describe the insertion of the subscapularis tendon on to the lesser tuberosity of the humerus. The transverse humeral ligament is also described at this level, as a band of tissue attached to the greater and lesser tuberosities, overlying the long tendon of biceps as it emerges from the capsule of the shoulder joint. The shoulder is a notorious site for anatomical variation but until recently little has been published with regard to the tendon of subscapularis. In this study, we illustrate that considerable variation in the insertion site of the tendon of subscapularis can be demonstrated using magnetic resonance imaging and that only 20% conform to the classic textbook description. In addition, a distinct transverse humeral ligament was identifiable in only a minority of shoulders examined (36%). PMID- 19306323 TI - Persistent reduction of conduction velocity and myelinated axon damage in vibrated rat tail nerves. AB - Prolonged hand-transmitted vibration exposure in the workplace has been recognized for almost a century to cause neurodegenerative and vasospastic disease. Persistence of the diseased state for years after cessation of tool use is of grave concern. To understand persistence of vibration injury, the present study examined recovery of nerve conduction velocity and structural damage of myelinated axons in a rat tail vibration model. Both 7 and 14 days of vibration (4 h/day) decreased conduction velocity. The decrease correlated directly with the increased percentage of disrupted myelinated axons. The total number of myelinated axons was unchanged. During 2 months of recovery, conduction velocity returned to control level after 7-day vibration but remained decreased after 14 day vibration. The rat tail model provides insight into understanding the persistence of neural deficits in hand-arm vibration syndrome. PMID- 19306324 TI - Iodothyronine deiodinases and thyroid hormone receptors regulation during flatfish (Solea senegalensis) metamorphosis. AB - Thyroid hormone-induced metamorphosis seems to represent an ancestral feature of chrordates (urochordates, cephalochordates and vertebrates), but also of nonchordate animals. Although thyroid hormones and thyroid hormone receptor profiles during metamorphosis have been analyzed in different vertebrate taxa, including fish, developmental expression and activity of type 2 (dio2, D2) and type 3 (dio3, D3) iodothyronine deiodinases, two key enzymes in anuran metamorphosis, remain unknown in any fish species. The aim of this work was to investigate the development of thyroid hormone system during the metamorphosis of a flatfish species, the Senegalese sole, focusing on the deiodinases developmental profile. We have cloned sole D2 and D3 and analyzed several parameters of thyroid hormones system in pre-, early-, middle-, and late metamorphic larvae. Both deiodinases contain in their catalytic centers an UGA triplet encoding for a selenocystein (Sec) residue as expected. Left eye migration and rotation in body position were associated with a significant increase in both thyroid hormones and thyroid hormone receptors at the middle late metamorphic stages. Although dio2 expression slightly increased during metamorphosis, D2 activity augmentation was much more significant. Sole dio3 expression declined only slightly, whereas the D3 activity clearly decreased at mid-late metamorphic period. This developmental profile of deiodinases sustained the rise of thyroid hormones levels observed during sole metamorphosis. No clear cut daily rhythms were observed in the parameters analyzed although it seemed that thyroid hormone system was more active during daytime, in particular at late metamorphic stages. These developmental changes point out the importance not only of thyroid hormones and their receptors but also of dio2 and dio3 in mediating flatfish metamorphosis, as it has been described in amphibians. PMID- 19306325 TI - Vibratory thresholds and mobility in older persons. AB - We tested the hypothesis that vibratory thresholds in the elderly are related to mobility. In all, 629 older persons without dementia underwent testing including 11 lower extremity performance measures and modified United Parkinson's Disease Rating Scale (UPDRS), summarized as composite mobility and global parkinsonian signs. Vibratory thresholds were measured at the ankle and toes bilaterally using the graduated Rydel-Seiffer tuning fork. In linear regression models adjusted for age, sex, and education, vibratory threshold was associated with composite mobility (estimate, 0.047, SE = 0.011, P < 0.001) and global parkinsonian signs score (estimate, -0.252, SE = 0.126, P = 0.047). These findings were primarily due to the association of vibratory threshold with gait and balance components of composite mobility and parkinsonian gait. These results were unchanged when we controlled for body mass index, physical activity, cognition, depression, vascular risk factors, vascular disease burden, joint pain, and falls. Vibratory thresholds are associated with mobility, supporting the link between peripheral sensory nerve function and mobility in the elderly. PMID- 19306326 TI - Time-frequency analysis of rhythmic masticatory muscle activity. AB - The aim of this study was to develop and validate under laboratory conditions an algorithm for a time-frequency analysis of rhythmic masticatory muscle activity (RMMA). The algorithm baseband demodulated the electromyographic (EMG) signal to provide a frequency versus time representation. Using appropriate thresholds for frequency and power parameters, it was possible to automatically assess the features of RMMA without examiner interaction. The algorithm was first tested using synthetic EMG signals and then using real EMG signals obtained from the masticatory muscles of 11 human subjects who underwent well-defined rhythmic, static, and possible confounding oral tasks. The accuracy of detection was quantified by receiver operating characteristics (ROC) curves. Sensitivity and specificity values were > or =90% and > or =96%, respectively. The areas under the ROC curves were > or =95% (standard error +/-0.1%). The proposed approach represents a promising tool to effectively investigate rhythmical contractions of the masticatory muscles. PMID- 19306327 TI - The increased incidence of the RET p.Gly691Ser variant in French-Canadian vesicoureteric reflux patients is not replicated by a larger study in Ireland. AB - The p.Gly691Ser variant of the RET protein, resulting from the 'A' allele of the SNP rs1799939 in exon 11 of the RET gene, was recently found to be present in a high proportion of primary vesicoureteric reflux (pVUR) patients in Quebec. We have determined the genotype of this SNP in 221 unrelated index cases of pVUR from the Irish population, in 190 full siblings of 160 of the index cases, and in 592 healthy controls. We found no significant difference in genotype or allele frequencies in patients and controls, and no tendency of affected siblings to share the same genotype. We also found no difference in the presence of additional phenotypic features such as duplex kidneys, between patients with and without the 'A' allele, and no difference in grade of reflux. We find no evidence of any influence of RET SNP rs1799939 on pVUR phenotype. PMID- 19306328 TI - Mutations of the EPHA2 receptor tyrosine kinase gene cause autosomal dominant congenital cataract. AB - Congenital cataracts (CCs) are clinically and genetically heterogeneous. Mutations in the same gene may lead to CCs differing in inheritance, morphology and severity. Loci for autosomal dominant posterior polar CC and total CC have both been mapped to the chromosomal 1p36 region harboring the EPHA2 receptor tyrosine kinase gene. Here, we report mutations of EPHA2 in three CC families from different ancestral groups. In a Chinese family with posterior polar CC, we identified a missense mutation, c.2819C>T (p.T940I), replacing a critical amino acid that functions at the receptor oligomerization interface. In a British family with posterior polar CC and an Australian family with total CC, we found a frameshift mutation (c.2915_2916delTG) and a splicing mutation (c.2826-9G>A), respectively. These two mutations are predicted to produce novel C-terminal polypeptides with 39 identical amino acids. Yeast two-hybrid analysis showed stronger interaction between the total CC-associated mutant EPHA2 and low molecular weight protein-tyrosine phosphatase, a negative regulator of EPHA2 signaling. Our results implicate the Eph-ephrin signaling system in development of human cataract and provide a novel insight into the molecular mechanism underlying the pathogenesis of human CCs. PMID- 19306330 TI - Rodney A. Appell. PMID- 19306331 TI - Two types of urgency. AB - AIMS: To determine whether urinary urgency, as defined by the International Continence Society, is an intensification of the normal sensation that occurs when micturition must be delayed once the urge to void is felt (Type 1 urgency) or a discrete, pathologic symptom different from the normal urge (Type 2 urgency). METHODS: Forty-eight consecutive patients who complained of urinary urgency completed two different questionnaires designed to answer the question posed above. The patients were divided into two groups of 24. For the test retest, group 1 completed questionnaire 1 twice within 3-10 days and group 2 did the same with questionnaire 2. On the second administration of the questionnaire, each subject crossed over and answered the other questionnaire. For the test retest, since the data set is dichotomous (yes/no), the degree of agreement between the two sets of data was assessed by calculating the kappa coefficient. RESULTS: There were 37 women and 11 men ranging in age from 54 to 87 years. There was no difference in age and sex between the two groups (P = 0.19). There was excellent agreement in the test-retest responses for both questionnaires (kappa = 1.0, P < 0.001). For questionnaire 1, the urge sensation was an intensification of the normal sensation in 33 (68.8%) and it was a different sensation in 15 (31.3%). Similarly, for questionnaire 2, it was an intensification of the normal urge in 34 (70.8%) and different in 14 (29.2%). The differences in patient responses between the two groups were not significant. In the crossover section, only 1 of 48 subjects changed their response, resulting in a very high degree of agreement (kappa = .95, p < .001). Combining the two groups, urgency was perceived as an intensification of the normal urge to void in 33/48 patients (69%), a different sensation in 14/48 (29%) and 1/48 (2%) was not sure. CONCLUSIONS: Urgency is comprised of at least two different sensations. One is an intensification of the normal urge to void and the other is a different sensation. The implications of this distinction are important insofar as they may have different etiologies and respond differently to treatment. PMID- 19306332 TI - Structural colored gels for tunable soft photonic crystals. AB - A periodically ordered interconnecting porous structure can be embodied in chemical gels by using closest-packed colloidal crystals as templates. The interconnecting porosity not only provides a quick response but also endows the porous gels with structural color arising from coherent Bragg optical diffraction. The structural colors revealed by porous gels can be regulated by several techniques, and thus, it is feasible to obtain desirable, smart, soft materials. A well-known thermosensitive monomer, N-isopropylacrylamide (NIPA), and other minor monomers were used to fabricate various structural colored gels. The selection of minor monomers depended on the targeted properties. This review focuses on the synthesis of templates, structural colored porous gels, and the applications of structural colored gel as smart soft materials for tunable photonic crystals. PMID- 19306333 TI - Bifunctional transition metal-based molecular catalysts for asymmetric C-C and C N bond formation. AB - This paper describes the recent advances in the conceptually new bifunctional Ir and Ru catalysts for asymmetric catalytic reactions. These reactions include the enantioselective Michael addition of 1,3-dicarbonyl compounds to cyclic enones and nitroalkenes, and the enantioselective direct amination of alpha cyanoacetates with diazoesters. The outcome of these reactions in terms of reactivity and selectivity was delicately influenced by the catalyst structures and the reaction conditions including the solvents used. Even with a 1 : 1 molar ratio of donors to acceptors, the reactions proceeded smoothly to give the corresponding chiral adducts with an excellent yield and enantiomeric excess (ee). Preliminary mechanistic studies showed that the key stage of the catalytic cycle is the interaction of the bifunctional catalyst with a pronucleophilic reagent that leads to stereoselective formation of C-, O-, or N-bound complexes. The resulting protonated catalyst bearing metal-bound nucleophiles readily reacts with electrophiles to provide C--C and C--N bond formation products in a highly stereoselective manner. PMID- 19306334 TI - Structural insights on pathogenic effects of novel mutations causing pyruvate carboxylase deficiency. AB - Pyruvate carboxylase (PC), a key enzyme for gluconeogenesis and anaplerotic pathways, consists of four domains, namely, biotin carboxylase (BC), carboxyltransferase (CT), pyruvate carboxylase tetramerization (PT), and biotin carboxyl carrier protein (BCCP). PC deficiency is a rare metabolic disorder inherited in an autosomal recessive way. The most severe form (form B) is characterized by neonatal lethal lactic acidosis, whereas patients with form A suffer chronic lactic acidosis with psychomotor retardation. Diagnosis of PC deficiency relies on enzymatic assay and identification of the PC gene mutations. To date, six mutations of the PC gene have been identified. We report nine novel mutations of the PC gene, in five unrelated patients: three being affected with form B, and the others with form A. Three of them were frameshift mutations predicted to introduce a premature termination codon, the remaining ones being five nucleotide substitutions and one in frame deletion. Impact of these mutations on mRNA was assessed by RT-PCR. Evidence for a deleterious effect of the missense mutations was achieved using protein alignments and three dimensional structural prediction, thanks to our modeling of the human PC structure. Altogether, our data and those previously reported indicate that form B is consistently associated with at least one truncating mutation, mostly lying in CT (C-terminal part) or BCCP domains, whereas form A always results from association of two missense mutations located in BC or CT (N-terminal part) domains. Finally, although most PC mutations are suggested to interfere with biotin metabolism, none of the PC-deficient patients was biotin-responsive. PMID- 19306335 TI - Interaction between a chromosome 10 RET enhancer and chromosome 21 in the Down syndrome-Hirschsprung disease association. AB - Individuals with Down syndrome (DS) display a 40-fold greater risk of Hirschsprung disease (HSCR) than the general population of newborns implicating chromosome 21 in HSCR etiology. Here we demonstrate that the RET enhancer polymorphism RET+9.7 (rs2435357:C>T) at chromosome 10q11.2 is associated with HSCR in DS individuals both by transmission disequilibrium (P=0.0015) and case control (P=0.0115) analysis of matched cases. Interestingly, the RET+9.7 T allele frequency is significantly different between individuals with DS alone (0.26+/ 0.04), HSCR alone (0.61+/-0.04), and those with HSCR and DS (0.41+/-0.04), demonstrating an association and interaction between RET and chromosome 21 gene dosage. This is the first report of a genetic interaction between a common functional variant (rs2435357) and a not infrequent copy number error (chromosome 21 dosage) in two human developmental disorders. PMID- 19306336 TI - Flexible host frameworks with diverse cavities in inclusion crystals of bile acids and their derivatives. AB - We have systematically investigated structures and properties of inclusion crystals of bile acids and their derivatives. These steroidal compounds form diverse host frameworks having zero-, one- and two-dimensional cavities, causing various inclusion behaviors towards many organic compounds. The diverse host frameworks exhibit the following guest-dependent flexibility. First, the frameworks mainly depend on the included guests in size and shape. The size dependence is quantitatively estimated by the parameter PCcavity, which is the volume ratio of a guest molecule to a host cavity. The resulting values of PCcavity lie in the range of 42-76%. Second, each of the host frameworks has its own range of the values. Some guests can employ two different frameworks with the boundary values, explaining formation of polymorphic crystals. Third, the host frameworks are selected by host-guest interactions through weak hydrogen bonds, such as NH/pi and CH/O. The weak hydrogen bonds play an important role for various selective inclusion processes. Fourth, the host frameworks are dynamically exchangeable, resulting in intercalation and polymerization in the cavities. These static and dynamic structures of the frameworks demonstrate great potential of crystalline organic inclusion compounds as functional materials. PMID- 19306337 TI - Synthesizing interlocked molecules dynamically. AB - As the complexity of mechanically interlocked molecular architectures increases, it is important to understand the underlying principles, such as molecular recognition and self-assembly processes, that govern the practice of template directed synthesis necessary to create these particular compounds. In this review, we explain the importance of dynamic processes in the synthesis of mechanically interlocked compounds. We show how many different dynamic covalent bonds have been used in the synthesis of rotaxanes, catenanes, and other higher order mechanically interlocked compounds, with the goal of revealing the state of the art in dynamic covalent chemistry. PMID- 19306338 TI - Physical activity in cancer survivors: a field in motion. AB - Physical activity (PA) is an important health behavior in almost any population but it may be particularly helpful for cancer survivors. OBJECTIVE: To introduce this special issue on PA in cancer survivors and to provide a summary of its important contributions to the field. METHODS: A brief historical review of PA research in cancer survivors followed by a narrative review of the articles published in this special issue. RESULTS: This special issue contains 13 original articles reporting 15 studies on PA in cancer survivors. Just over half of the studies focus on breast cancer survivors, whereas the remainder focus on understudied cancer survivor groups such as lung, ovarian, colorectal, prostate, hematologic, and pediatric. Moreover, a majority of the studies focus on the survivorship phase of the cancer continuum. Perhaps the most distinctive feature of this special issue is the number of studies focusing on the determinants of PA. Taken together, the 13 articles make significant contributions in four areas: (1) randomized controlled trials of PA interventions with supportive care endpoints, (2) observational studies on the determinants of PA, (3) observational studies on the 'determinants of PA determinants', and (4) studies on methodological and feasibility issues related to conducting PA trials. CONCLUSIONS: PA research is making an important contribution to the health and well-being of cancer survivors across the entire cancer control continuum. This special issue builds on this momentum and provides the single largest collective contribution of knowledge to date in the field of PA in cancer survivors. PMID- 19306339 TI - Influence of oxygen and carbogen breathing on renal oxygenation measured by T2* weighted imaging at 3.0 T. AB - The aim of the study was to assess the influence of carbogen (95% O(2), 5% CO(2)) or pure oxygen breathing on renal oxygenation measured by blood oxygenation level dependent (BOLD) magnetic resonance imaging at 3.0 T. Seven healthy young volunteers (median age 25, range 23-35 years) participated in the study. A T2* weighted fat-saturated spoiled gradient-echo sequence was implemented on a 3.0 T whole-body imager (TE/TR = 27.9 ms/49 ms, excitation angle 20 degrees ) with an acquisition time of approximately 5.3 s. A total of 100 images were acquired during 22 min. A block design was applied for gas administration: 4 min room air, 4 min carbogen/oxygen, 4 min room air, 4 min carbogen/oxygen and 6 min room air. A compartment model was fitted to the data sets accounting for time-dependent increase/decrease of renal oxygenation as well as baseline changes of the scanner. T2*-weighted images showed good image quality without notable artefacts or distortions. Mean relative signal increase due to carbogen breathing was 2.73% (95% confidence interval: 1.34-5.54) in the right kidney and 3.76% (1.53-9.20) in the left kidney, while oxygen breathing led to a signal enhancement of 3.20% (2.57-3.98) in the right kidney and 3.16% (1.83-5.45) in the left kidney. No statistical difference was found between carbogen and oxygen breathing or between the oxygenation of the right and the left kidney. A significant difference was found in the characteristic time constant for the signal increase with a faster saturation taking place for oxygen breathing. Renal tissue oxygenation is clearly influenced by carbogen or oxygen breathing. The changes can be assessed by T2* weighted MRI at high field strengths. The effects are in the expected range for the BOLD effect of 3-4% at 3.0 T. The proposed technique might be interesting for the assessment of renal tissue oxygenation and its regulation in patients with kidney diseases. PMID- 19306340 TI - Diffusion-weighted MRI measurements on stroke patients reveal water-exchange mechanisms in sub-acute ischaemic lesions. AB - The aim of this study was to investigate the diffusion time dependence of signal versus-b curves obtained from diffusion-weighted magnetic resonance imaging (DW MRI) of sub-acute ischaemic lesions in stroke patients. In this case series study, 16 patients with sub-acute ischaemic stroke were examined with DW-MRI using two different diffusion times (60 and 260 ms). Nine of these patients showed sufficiently large lesions without artefacts to merit further analysis. The signal-versus-b curves from the lesions were plotted and analysed using a two compartment model including compartmental exchange. To validate the model and to aid the interpretation of the estimated model parameters, Monte Carlo simulations were performed. In eight cases, the plotted signal-versus-b curves, obtained from the lesions, showed a signal-curve split-up when data for the two diffusion times were compared, revealing effects of compartmental water exchange. For one of the patients, parametric maps were generated based on the extracted model parameters. These novel observations suggest that water exchange between different water pools is measurable and thus potentially useful for clinical assessment. The information can improve the understanding of the relationship between the DW-MRI signal intensity and the microstructural properties of the lesions. PMID- 19306341 TI - Construct optimization for protein NMR structure analysis using amide hydrogen/deuterium exchange mass spectrometry. AB - Disordered or unstructured regions of proteins, while often very important biologically, can pose significant challenges for resonance assignment and three dimensional structure determination of the ordered regions of proteins by NMR methods. In this article, we demonstrate the application of (1)H/(2)H exchange mass spectrometry (DXMS) for the rapid identification of disordered segments of proteins and design of protein constructs that are more suitable for structural analysis by NMR. In this benchmark study, DXMS is applied to five NMR protein targets chosen from the Northeast Structural Genomics project. These data were then used to design optimized constructs for three partially disordered proteins. Truncated proteins obtained by deletion of disordered N- and C-terminal tails were evaluated using (1)H-(15)N HSQC and (1)H-(15)N heteronuclear NOE NMR experiments to assess their structural integrity. These constructs provide significantly improved NMR spectra, with minimal structural perturbations to the ordered regions of the protein structure. As a representative example, we compare the solution structures of the full length and DXMS-based truncated construct for a 77-residue partially disordered DUF896 family protein YnzC from Bacillus subtilis, where deletion of the disordered residues (ca. 40% of the protein) does not affect the native structure. In addition, we demonstrate that throughput of the DXMS process can be increased by analyzing mixtures of up to four proteins without reducing the sequence coverage for each protein. Our results demonstrate that DXMS can serve as a central component of a process for optimizing protein constructs for NMR structure determination. PMID- 19306342 TI - Graphical models of protein-protein interaction specificity from correlated mutations and interaction data. AB - Protein-protein interactions are mediated by complementary amino acids defining complementary surfaces. Typically not all members of a family of related proteins interact equally well with all members of a partner family; thus analysis of the sequence record can reveal the complementary amino acid partners that confer interaction specificity. This article develops methods for learning and using probabilistic graphical models of such residue "cross-coupling" constraints between interacting protein families, based on multiple sequence alignments and information about which pairs of proteins are known to interact. Our models generalize traditional consensus sequence binding motifs, and provide a probabilistic semantics enabling sound evaluation of the plausibility of new possible interactions. Furthermore, predictions made by the models can be explained in terms of the underlying residue interactions. Our approach supports different levels of prior knowledge regarding interactions, including both one-to one (e.g., pairs of proteins from the same organism) and many-to-many (e.g., experimentally identified interactions), and we present a technique to account for possible bias in the represented interactions. We apply our approach in studies of PDZ domains and their ligands, fundamental building blocks in a number of protein assemblies. Our algorithms are able to identify biologically interesting cross-coupling constraints, to successfully identify known interactions, and to make explainable predictions about novel interactions. PMID- 19306343 TI - Complexes of HIV-1 integrase with HAT proteins: multiscale models, dynamics, and hypotheses on allosteric sites of inhibition. AB - A new and very promising strategy for HIV drug discovery consists in blocking the multiple functional interactions between HIV-1 integrase (IN) and its cellular cofactors. At present, this line of action is hindered by the absence of three dimensional structures of IN in complex with any of them. In this article, we developed a full-length three-dimensional structure of IN, including the highly flexible terminal residues 270-288, which are not experimentally solved. Additionally, we built models of IN complexed to the human acetyltransferases GCN5 and p300 based on available structural and mutagenesis data. Then, we studied the dynamical behavior of these models by means of the Coarse-Grained Molecular Dynamics (CGMD) and Essential Dynamics (ED) to locate and characterize the nature of the largest collective motions. We found correlated motions involving distant regions of IN. Moreover, we found that these are influenced by the binding with the acetyltransferases (HATs). Taken together these findings suggest a way to affect the acetyltransferase binding by an allosteric type of inhibition and provide an important new approach for the drug design against HIV disease. PMID- 19306344 TI - Recent developments in testicular germ cell tumor research. AB - Testicular germ cell tumors of adolescents and adults (TGCTs; the so-called type II variant) are the most frequent malignancies found in Caucasian males between 20 and 40 years of age. The incidence has increased over the last decades. TGCTs are divided into seminomas and nonseminomas, the latter consisting of the subgroups embryonal carcinoma, yolk-sac tumor, teratoma, and choriocarcinoma. The pathogenesis starts in utero, involving primordial germ cells/gonocytes that are blocked in their differentiation, and develops via the precursor lesion carcinoma in situ toward invasiveness. TGCTs are totipotent and can be considered as stem cell tumors. The developmental capacity of their cell of origin, the primordial germ cells/gonocyte, is demonstrated by the different tumor histologies of the invasive TGCTs. Seminoma represents the germ cell lineage, and embryonal carcinoma is the undifferentiated component, being the stem cell population of the nonseminomas. Somatic differentiation is seen in the teratomas (all lineages), whereas yolk-sac tumors and choriocarcinoma represent extra-embryonal differentiation. Seminomas are highly sensitive to irradiation and (DNA damaging) chemotherapy, whereas most nonseminomatous elements are less susceptible to radiation, although still sensitive to chemotherapy, with the exception of teratoma. To allow early diagnosis and follow up, appropriate markers are mandatory to discriminate between the different subgroups. In this review, a summary will be given related to several recent developments in TGCT research, especially selected because of their putative clinical impact. PMID- 19306345 TI - Spermatogonial stem cells: mouse and human comparisons. AB - Spermatogonial stem cells (SSCs) have unique characteristics in that they produce sperm that transmit genetic information from generation to generation and they can be reprogrammed spontaneously to form embryonic stem (ES)-like cells to acquire pluripotency. In rodents, it is generally believed that the A-single (A(s)) is the stem cell population, whereas the A-paired (A(pr)) and A-aligned (A(al)) represent the progenitor spermatogonial population. The A(1) to A(4) cells, intermediate, and type B spermatogonia are considered differentiated spermatogonia. In human, very little information is available about SSCs, except for the earlier work of Clermont and colleagues who demonstrated that there are two different types of A spermatogonia, the A(dark) and A(pale) spermatogonia. The A(dark) spermatogonia were referred to as the reserve stem cells, whereas the A(pale) were considered the renewing stem cells. In this review, we outline several spermatogonial renewal schemes for both rodents and primates, including man. We also compare phenotypic markers for spermatogonia/spermatogonial stem cells in rodents and humans and address SSC potential and therapeutic application. PMID- 19306346 TI - Gonocytes, the forgotten cells of the germ cell lineage. AB - Male germ cells, the repository cells of the genome, comprise several successive developmental stages starting in the embryo and ending up with the spermatozoon. Gonocytes represent the fetal and neonatal stages preceding the formation of spermatogonial stem cells. Recent findings shows that germline stem cells can be driven to pluripotency and used as alternative for embryonic stem cells prompted more effort in identifying the processes regulating the development of their precursors, the testicular gonocytes. Also called pre- or pro-spermatogonia, gonocytes represent not one, but several successive developmental stages between the time at which the germ cell becomes resident in the forming fetal testis to the time it migrates to the basement membrane of the seminiferous cord to adopt a spermatogonial phenotype. This review summarizes the findings regarding the genetic identity of gonocytes, providing a description of the "common" gene expression profiles of fetal and neonatal gonocytes, as well as information on the main regulatory factors of gonocyte functions. A better comprehension of gonocyte development should help in the understanding of how germline stem cells are formed, possibly providing valuable clues on the origins of germ cell tumors or infertility. PMID- 19306347 TI - Current knowledge in the renewal capability of germ cells in the adult ovary. AB - It is a central dogma in reproductive biology that oogenesis is completed before or just after birth and that the postnatal ovary is endowed by a fixed and non renewing number of oocytes in mammals. However, this widely accepted doctrine was recently challenged by studies showing regeneration of oocytes from putative germ cells in bone marrow and peripheral blood. These results not only triggered an enormous amount of interest among reproductive biologists but also a great deal of debate. In this review we will provide an update on the molecular aspects of the formation of primordial germ cells (PGC), the precursors of adult gametocytes, beginning from their specification to their migration to prospective gonads and formation of the ovary and follicular structures. We will also discuss more recent studies that showed in vivo regeneration of germ cells in the postnatal ovary in situ, along with other pioneering works that demonstrated generation of germ cells in vitro from embryonic and somatic stem cells. PMID- 19306348 TI - Gonadoblastoma locus and the TSPY gene on the human Y chromosome. AB - The gonadoblastoma (GBY) locus is the only oncogenic locus on the human Y chromosome. It is postulated to serve a normal function in the testis, but could exert oncogenic effects in dysgenetic gonads of individuals with intersex and/or dysfunctional testicular phenotypes. Recent studies establish the testis-specific protein Y-encoded (TSPY) gene to be the putative gene for GBY. TSPY serves normal functions in male stem germ cell proliferation and differentiation, but is ectopically expressed in early and late stages of gonadoblastomas, testicular carcinoma in situ (the premalignant precursor for all testicular germ cell tumors), seminomas, and selected nonseminomas. Aberrant TSPY expression stimulates protein synthetic activities, accelerates cell proliferation, and promotes tumorigenicity in athymic mice. TSPY binds to type B cyclins, enhances an activated cyclin B-CDK1 kinase activity, and propels a rapid G(2)/M transition in the cell cycle. TSPY also counteracts the normal functions of its X-homologue, TSPX, which also binds to cyclin B and modulates the cyclin B-CDK1 activity to insure a proper G(2)/M transition in the cell cycle. Hence, ectopic expression and actions of the Y-located TSPY gene in incompatible germ cells, such as those in dysgenetic or ovarian environments and dysfunctional testis, disrupt the normal cell cycle regulation and predispose the host cells to tumorigenesis. The contrasting properties of TSPY and TSPX suggest that somatic cancers, such as intracranial germ cell tumors, melanoma, and hepatocellular carcinoma, with detectable TSPY expression could exhibit sexual dimorphisms in the initiation and/or progression of the respective oncogenesis. PMID- 19306349 TI - Signaling pathways in spermatogonial stem cells and their disruption by toxicants. AB - Spermatogenesis is a complex biological process that is particularly sensitive to environmental insults such as chemicals and physical stressors. Exposure to specific chemicals has been shown to inhibit fertility through a negative impact on germ cell proliferation and differentiation that can lower sperm count. In addition, toxicants might produce DNA damages that could have negative consequences on the development of the offspring. This review describes spermatogonial stem cell development in the testis, signaling pathways that are crucial for self-renewal, and possible target molecules for environmental toxicants such as phthalate esters and nanoparticles. PMID- 19306350 TI - Immune physiology and oogenesis in fetal and adult humans, ovarian infertility, and totipotency of adult ovarian stem cells. AB - It is still widely believed that while oocytes in invertebrates and lower vertebrates are periodically renewed throughout life, oocytes in humans and higher vertebrates are formed only during the fetal/perinatal period. However, this dogma is questioned, and clashes with Darwinian evolutionary theory. Studies of oogenesis and follicular renewal from ovarian stem cells (OSCs) in adult human ovaries, and of the role of third-party bone marrow-derived cells (monocyte derived tissue macrophages and T lymphocytes) could help provide a better understanding of the causes of ovarian infertility, its prevention, and potential treatment. We have reported differentiation of distinct cell types from OSC and the production of new eggs in cultures derived from premenopausal and postmenopausal human ovaries. OSCs are also capable of producing neural/neuronal cells in vitro after sequential stimulation with sex steroid combinations. Hence, OSC represent a unique type of totipotent adult stem cells, which could be utilized for autologous treatment of premature ovarian failure and also for autologous stem cell therapy of neurodegenerative diseases without use of allogeneic embryonic stem cells or somatic cell nuclear transfer. The in vivo application of sex steroid combinations may augment the proliferation of existing neural stem cells and their differentiation into mature neuronal cells (systemic regenerative therapy). Such treatment may also stimulate the transdifferentiation of autologous neural stem cell precursors into neural stem cells useful for topical or systemic regenerative treatment. PMID- 19306352 TI - Integrative high-resolution microarray analysis of human myeloma cell lines reveals deregulated miRNA expression associated with allelic imbalances and gene expression profiles. AB - It is thought that altered microRNA (miRNA) expression due to various mechanisms plays a critical role in the pathogenesis of most human cancers. Notably, about half of the known miRNAs are intragenic and frequently coexpressed with their host genes. To date there is little evidence concerning miRNA expression in multiple myeloma (MM). In an attempt to provide insights into miRNA deregulation in MM, we profiled global miRNA expression in a panel of molecularly well characterized human myeloma cell lines (HMCLs) using high-resolution microarrays, and then used integrative analyses to identify altered patterns correlated with DNA copy number (CN) or gene expression profiles. We identified 16 miRNAs mapped to chromosomal regions frequently involved in numerical imbalances in MM, whose expression significantly correlated with the CN of the corresponding miRNA genes; among these, miR-22 expression was also affected by chromosome arm 17p loss in a representative panel of primary MM tumors. The expression of 32 intronic miRNAs significantly correlated with that of their host transcripts, some of which were highly deregulated in MM patients. The expression of some of the miRNAs was validated by quantitative RT-PCR. Finally, a number of the identified miRNAs have previously been reported to play important roles in tumorigenesis. Overall, our data highlight that genomic alterations may significantly affect miRNA expression in HMCLs and demonstrate a frequent coexpression of intronic miRNAs with their host genes that may have a pathogenetic relevance in plasma cell transformation. PMID- 19306353 TI - Centrioles to basal bodies in the spermiogenesis of Mastotermes darwiniensis (Insecta, Isoptera). AB - In addition to their role in centrosome organization, the centrioles have another distinct function as basal bodies for the formation of cilia and flagella. Centriole duplication has been reported to require two alternate assembly pathways: template or de novo. Since spermiogenesis in the termite Mastotermes darwiniensis lead to the formation of multiflagellate sperm, this process represents a useful model system in which to follow basal body formation and flagella assembly. We present evidence of a possible de novo pathway for basal body formation in the differentiating germ cell. This cell also contains typical centrosomal proteins, such as centrosomin, pericentrin-like protein, gamma tubulin, that undergo redistribution as spermatid differentiation proceeds. The spermatid centrioles are long structures formed by nine doublet rather than triplet microtubules provided with short projections extending towards the surrounding cytoplasm and with links between doublets. The sperm basal bodies are aligned in parallel beneath the nucleus. They consist of long regions close to the nucleus showing nine doublets in a cartwheel array devoid of any projections; on the contrary, the short region close to the plasma membrane, where the sperm flagella emerge, is characterized by projections similar to those observed in the centrioles linking the basal body to the plasma membrane. It is hypothesized that this appearance is in connection with the centriole elongation and further with the flagellar axonemal organization. Microtubule doublets of sperm flagellar axonemes are provided with outer dynein arms, while inner arms are rarely visible. PMID- 19306354 TI - Cyclosporine therapy in refractory/relapsed patients with thrombotic thrombocytopenic purpura. PMID- 19306351 TI - Genomic landscape of developing male germ cells. AB - Spermatogenesis is a highly orchestrated developmental process by which spermatogonia develop into mature spermatozoa. This process involves many testis- or male germ cell-specific gene products whose expressions are strictly regulated. In the past decade the advent of high-throughput gene expression analytical techniques has made functional genomic studies of this process, particularly in model animals such as mice and rats, feasible and practical. These studies have just begun to reveal the complexity of the genomic landscape of the developing male germ cells. Over 50% of the mouse and rat genome are expressed during testicular development. Among transcripts present in germ cells, 40% - 60% are uncharacterized. A number of genes, and consequently their associated biological pathways, are differentially expressed at different stages of spermatogenesis. Developing male germ cells present a rich repertoire of genetic processes. Tissue-specific as well as spermatogenesis stage-specific alternative splicing of genes exemplifies the complexity of genome expression. In addition to this layer of control, discoveries of abundant presence of antisense transcripts, expressed psuedogenes, non-coding RNAs (ncRNA) including long ncRNAs, microRNAs (miRNAs) and Piwi-interacting RNAs (piRNAs), and retrogenes all point to the presence of multiple layers of expression and functional regulation in male germ cells. It is anticipated that application of systems biology approaches will further our understanding of the regulatory mechanism of spermatogenesis. PMID- 19306355 TI - First-line therapy for chronic myeloid leukemia: Past, present, and future. AB - The development of Bcr-Abl tyrosine kinase inhibitors has dramatically changed the prognosis of patients with newly diagnosed chronic myeloid leukemia (CML). Standard-dose imatinib (400 mg/day in chronic phase, 600 mg/day in advanced CML) now dominates the management of this disease, producing considerably higher hematologic, cytogenetic, and molecular response rates than seen with previous drug therapies. However, although many patients respond well to standard-dose imatinib initially, some patients do not achieve adequate levels of response or discontinue therapy because of resistance. One approach to improving treatment response with first-line imatinib may be to increase the imatinib dose (800 mg/day), although recent trial data indicate that overall increases in response rates may be modest. Newer Bcr-Abl tyrosine kinase inhibitors can induce responses in patients with all phases of imatinib-resistant CML, even those with imatinib-resistant mutations in the BCR-ABL gene. Furthermore, in initial studies, first-line dasatinib or nilotinib treatment has produced response rates that compare favorably with historical controls treated with imatinib, although confirmation is required from head-to-head clinical trials. Future clinical approaches may include drug combinations, which may allow quiescent leukemia stem cells to be eradicated. Further improvements in drug treatment for first-line CML are expected during the next few years. PMID- 19306356 TI - AML/MDS with 11q/MLL amplification show characteristic gene expression signature and interplay of DNA copy number changes. AB - AML/MDS patients carrying 11q amplifications involving the mixed lineage leukemia gene (MLL) locus are characterized by a complex aberrant karyotype (CAK) frequently including deletions within 5q, 17p, and 7q, older age and fast progression of the disease with extremely poor prognosis. MLL has been shown to be overexpressed in cases with 11q amplification. However, in most of the cases, the amplified region is not restricted to the MLL locus. In this study, we investigated 19 patients with AML/MDS and MLL gain/amplification. By means of array CGH performed in 12 patients, we were able to delineate the minimal deleted regions within 5q and 17p and identified three independent regions 11q/I-III that were amplified in all cases. Gene expression profiles established in 15 cases were used to identify candidate genes within these regions. Notably, analysis of our data suggests a correlation of loss of 5q and 17p and expression of genes present in 11q23-25. Furthermore, we demonstrate that the gene expression signature can be used to discriminate AML/MDS with MLL amplification from several other types of AML. PMID- 19306357 TI - Unusual complication of deep brain stimulation in Parkinson's disease. PMID- 19306358 TI - Microglia and neuropathic pain. AB - In contrast to physiological pain, pathological pain is not dependent on the presence of tissue-damaging stimuli. One type of pathological pain--neuropathic pain--is often a consequence of nerve injury or of diseases such as diabetes, AIDS, or cancer. Neuropathic pain can be agonizing, can persist over long periods, and, unfortunately, is often resistant to known painkillers. There is a rapidly growing body of evidence indicating that microglia, the CNS immune cells, have causal roles in the pathogenesis of pain hypersensitivity following nerve injury. We will review recent advances in our understanding of the mechanisms producing neuropathic pain, focusing on the roles of microglia-expressed molecules, including cell surface receptors, intracellular signaling molecules, and diffusible factors involved in nerve injury-induced pain behaviors and hyperexcitability of dorsal horn neurons. Elucidating how spinal microglia cause neuropathic pain may provide us with exciting insights into pain mechanisms and clues for developing new drugs for the treatment of neuropathic pain. PMID- 19306359 TI - IL-12 p40 homodimer, the so-called biologically inactive molecule, induces nitric oxide synthase in microglia via IL-12R beta 1. AB - Earlier we have demonstrated that IL-12 p40 homodimer (p40(2)) induces the expression of inducible nitric oxide synthase (iNOS) in microglia. This study was undertaken to investigate underlying mechanisms required for IL-12 p40(2)- and IL 12 p70-induced expression of iNOS in microglia. IL-12 p40(2) alone induced the activation of both extracellular signal-regulated kinase (ERK) and p38 mitogen activated protein kinase (MAPK). Interestingly, the ERK pathway coupled p40(2) to iNOS expression via C/EBP beta, but not NF-kappaB, whereas the p38 pathway relayed the signal from p40(2) to iNOS expression via both NF-kappaB and C/EBP beta. Furthermore, by using microglia from IL-12R beta 1 (-/-) and IL-12R beta 2 (-/-) mice or siRNA against IL-12R beta 1 and IL-12R beta 2, we demonstrate that p40(2) induced the expression of iNOS in microglia via IL-12R beta 1-(ERK+p38) (NF-kappaB +C/EBP beta) pathway. In contrast, both IL-12R beta 1 and IL-12R beta 2 were involved for IL-12 p70-induced microglial expression of iNOS. Although IL 12R beta 1 coupled p70 to NF-kappaB and C/EBP beta, IL-12R beta 2 was responsible for p70-mediated activation of GAS. This study delineates a new role of IL-12R beta 1 and IL-12R beta 2 for the expression of iNOS and production of NO in microglia that may participate in the pathogenesis of neuroinflammatory diseases. PMID- 19306360 TI - Mitogen-activated protein kinase-signaling regulates the ability of Muller glia to proliferate and protect retinal neurons against excitotoxicity. AB - The purpose of this study was to investigate whether insulin, fibroblast growth factor (FGF), and mitogen-activated protein kinase (MAPK) pathways protect retinal neurons against excitotoxicity and regulate the proliferation of Muller glia. We found that intraocular injections of insulin or FGF2 had variable effects upon the phosphorylation of ERK1/2, p38 MAPK, and CREB, and the expression of immediate early genes, cFos and Egr1. Accumulations of pERK1/2, p38 MAPK, pCREB, cFos and Egr1 in response to insulin or FGF2 were confined to Muller glia, whereas retinal neurons did not seem to respond to growth factors. Unlike FGF2, insulin stimulated microglia-like cells to upregulate the intermediate filament transitin and lysosomal membrane glycoprotein (LMG). With microglia-like cells and Muller glia stimulated by insulin or FGF2 there were profound effects upon numbers of dying neurons in response to excitotoxic damage. Although FGF2 significantly reduced numbers of dying neurons, insulin significantly increased numbers of dying neurons. In addition to neuroprotective affects, FGF2 also "primed" the Muller glia to proliferate following retinal damage, whereas insulin had no effect upon glial proliferation. Further, we found that FGF receptor isoform 1 (FGFR1) and FGFR3 were prominently expressed in the retina, whereas the insulin receptor and FGFR2 are not expressed, or are expressed at very low levels. We conclude that MAPK-signaling through FGF receptors stimulates Muller glia to become more neuroprotective and progenitor-like, whereas insulin acting on Muller and microglia-like cells through unidentified receptors had the opposite effect. PMID- 19306361 TI - Progression of conduction system disease in a paced patient with Kearns-Sayre syndrome. AB - The Kearns-Sayre syndrome is a rare condition characterized by external ophthalmoplegia, retinal pigmentary degeneration and progressive impairment of cardiac conduction. We report a patient with the permanent pacemaker. The spontaneous cardiac impulse formation disappeared during long-term follow-up in this patient with Kearns-Sayre Syndrome. PMID- 19306362 TI - Proposed neuroimaging criteria for the diagnosis of multiple system atrophy. AB - In this article, we review the state of the art knowledge concerning structural and functional imaging in multiple system atrophy (MSA). The relative value of imaging modalities in the differential diagnosis of MSA from other parkinsonian syndromes is debated. It is concluded that, although neuroimaging biomarkers provide valuable supportive data alongside clinical assessments, it is not possible to use them as surrogate markers. PMID- 19306363 TI - fMRI activation mapping as a percentage of local excitation: consistent presurgical motor maps without threshold adjustment. AB - PURPOSE: To evaluate the performance of a relative activation amplitude algorithm, versus standard t-value thresholding, for reliably establishing the location, amplitude, and spatial extent of functional magnetic resonance imaging (fMRI) brain activation for presurgical planning. MATERIALS AND METHODS: Diagnostic fMRI maps from 42 neurosurgical patients performing a simple hand movement task were analyzed. Relative activation maps were made by normalizing statistical t-value maps to the local peak activation amplitude within each functional brain region. The spatial distribution of activation was quantified and compared across mapping algorithms, subjects, and scan duration. RESULTS: Whereas the spatial distribution of blood oxygenation level-dependent (BOLD) t value statistical activation maps was highly variable across subjects and scan duration, the spatial distribution of relative activation maps was highly reproducible both within individual subjects and across different subjects. In every case the 40% most active voxels in the cortical hand region were consistently localized to the pre- and postcentral gyri of the sensorimotor cortex. CONCLUSION: The reproducibility and anatomical specificity of the spatiotemporal pattern of BOLD activation makes relative amplitude fMRI mapping a useful tool for clinical imaging, where accuracy, reproducibility, and quality control are critical concerns. PMID- 19306364 TI - Multicenter, double-blind, randomized, intra-individual crossover comparison of gadobenate dimeglumine and gadopentetate dimeglumine in MRI of brain tumors at 3 tesla. AB - PURPOSE: To prospectively compare 0.1 mmol/kg doses of gadobenate dimeglumine and gadopentetate dimeglumine for contrast-enhanced MRI of brain lesions at 3 Tesla (T). MATERIALS AND METHODS: Forty-six randomized patients underwent a first examination with gadobenate dimeglumine (n = 23) or gadopentetate dimeglumine (n = 23) and then, after 2-7 days, a second examination with the other agent. Contrast administration (volume, rate), sequence parameters (T1wSE; T1wGRE), and interval between injection and image acquisition were identical for examinations in each patient. Three blinded neuroradiologists evaluated images qualitatively (lesion delineation, lesion enhancement, global preference) and quantitatively (lesion-to-brain ratio [LBR], contrast-to-noise ratio [CNR], % lesion enhancement). Differences were assessed using Wilcoxon's signed-rank test. Reader agreement was determined using kappa (kappa) statistics. RESULTS: There were no demographic differences between groups. The three readers preferred gadobenate dimeglumine globally in 22 (53.7%), 21 (51.2%), and 27 (65.9%) patients, respectively, compared with 0, 1, and 0 patients for gadopentetate dimeglumine. Similar significant (P < 0.001) preference was expressed for lesion border delineation and enhancement. Reader agreement was consistently good (kappa = 0.48 0.64). Significantly (P < 0.05) higher LBR (+43.5- 61.2%), CNR (+51.3-147.6%), and % lesion enhancement (+45.9-49.5%) was noted with gadobenate dimeglumine. CONCLUSION: Brain lesion depiction at 3T is significantly improved with 0.1 mmol/kg gadobenate dimeglumine. PMID- 19306365 TI - Clinical correlations of brain lesion distribution in multiple sclerosis. AB - PURPOSE: To explore relations between spatial distribution of multiple sclerosis (MS) lesions, and disability. In MS, the presence of asymptomatic brain lesions challenges the prediction of disability based on conventional brain MRI. Hypothesizing that symptomatology may partly be determined by lesion location, this retrospective study explored relations between lesion location and disability using voxelwise analyses in standard space. MATERIALS AND METHODS: Using nonparametric permutation-based statistics, voxelwise lesion probability on T2 lesion masks was related to expanded disability status scale (EDSS) and MS functional composite (MSFC) subdomain scores and demographic characteristics of 325 MS patients. To identify statistically significant locations, a cluster forming threshold of 3.1 was used. RESULTS: In clusters in the periventricular region, lesion probability correlated significantly (P < 0.001) with disability and disease duration, and was higher in progressive than in relapsing disease. When controlled for lesion load (LL), no significant clusters survived. Presence and number of spinal cord lesions did not correlate with lesion probability in any location, and did not influence correlations with disability when included in its analyses. CONCLUSION: Periventricular lesions were related to disability. LL influenced relations between disability and lesion probability throughout the brain, suggesting interplay between lesional burden and its location in determining disability in MS. PMID- 19306366 TI - Use of ultrasmall superparamagnetic particles of iron oxide (USPIO)-enhanced MRI to demonstrate diffuse inflammation in the normal-appearing white matter (NAWM) of multiple sclerosis (MS) patients: an exploratory study. AB - PURPOSE: To explore ultrasmall superparamagnetic particles of iron oxide (USPIO) as a marker for diffuse inflammation in multiple sclerosis (MS) normal-appearing white matter (NAWM), using quantitative MRI. Disease activity in the NAWM of MS patients partly explains why MRI lesion burden correlates only moderately with disability. USPIO have been shown to visualize the cellular component of inflammation in focal MS lesions. In this study, we aimed to explore USPIO as a marker for the more diffuse inflammation in MS NAWM, using quantitative MRI. MATERIALS AND METHODS: In this prospective MRI study, 16 MS patients (eight relapsing-remitting MS [RRMS] and eight primary-progressive MS [PPMS] cases) and five healthy control (HC) subjects were included. Using a flip-angle (FA) array, B1-corrected T1 maps were generated before and 24 hours after USPIO (SHU555C) injection. White-matter (WM) T1 histogram and region-of-interest (ROI) characteristics were compared between both time points using Wilcoxon signed-rank test. RESULTS: Both NAWM ROI and histogram analyses showed T1 shortening after USPIO injection in MS patients (P < 0.01), but not in HCs (P = 0.68). CONCLUSION: This exploratory study suggests that USPIO-enhanced MRI may be a new potential marker for subtle inflammatory activity in MS NAWM. Further studies should focus on relating diffuse inflammation to clinical disease activity and treatment efficacy. PMID- 19306367 TI - Enhancement of neuronal outward delayed rectifier K+ current by human monocyte derived macrophages. AB - Macrophages are critical cells in mediating the pathology of neurodegenerative disorders and enhancement of neuronal outward potassium (K(+)) current has implicated in neuronal apoptosis. To understand how activated macrophages induce neuronal dysfunction and injury, we studied the effects of lipopolysaccharide (LPS)-stimulated human monocytes-derived macrophage (MDM) on neuronal outward delayed rectifier K(+) current (I(K)) and resultant change on neuronal viability in primary rat hippocampal neuronal culture. Bath application of LPS-stimulated MDM-conditioned media (MCM) enhanced neuronal I(K) in a concentration-dependent manner, whereas non-stimulated MCM failed to alter neuronal I(K). The enhancement of neuronal I(K) was repeated in a macrophage-neuronal co-culture system. The link of stimulated MCM (MCM(+))-associated enhancement of I(K) to MCM(+)-induced neuronal injury, as detected by PI/DAPI (propidium iodide/4',6-diamidino-2 phenylindol) staining and MTT assay, was demonstrated by experimental results showing that addition of I(K) blocker tetraethylammonium to the culture protected hippocampal neurons from MCM(+)-associated challenge. Further investigation revealed elevated levels of K(v) 1.3 and K(v) 1.5 channel expression in hippocampal neurons after addition of MCM(+) to the culture. These results suggest that during brain inflammation macrophages, through their capacity of releasing bioactive molecules, induce neuronal injury by enhancing neuronal I(K) and that modulation of K(v) channels is a new approach to neuroprotection. PMID- 19306368 TI - Management of traumatic brain injury: first link in chain of survival. AB - Traumatic brain injury is a significant cause of morbidity and mortality. The prehospital care of the patient with a traumatic brain injury is critical to maximizing the chances for a good outcome. Prehospital management of the traumatic brain injury patient is directed toward preventing and limiting secondary brain injury while facilitating rapid transport to an appropriate facility capable of providing definitive neurocritical care. Key points in management include the assessment of oxygenation, blood pressure, and mental status (as measured with the Glasgow Coma Scale) and the pupillary examination. Treatment strategies are directed toward maintaining adequate oxygenation and perfusion and treating herniation. Judicious use of temporary hyperventilation and hypertonic saline are considerations. This review provides the most recent evidence regarding the treatment of traumatic brain injury in the prehospital setting and introduces areas in need of future research. PMID- 19306369 TI - Severe traumatic brain injury: maximizing outcomes. AB - Severe traumatic brain injury is one of the leading causes of death and disability in the United States. The initial management of traumatic brain injury involves early resuscitation, computed tomography scanning, and surgical evacuation of mass lesions, when indicated. Recent research suggests that the prevention and treatment of secondary brain injury decrease mortality and improve outcomes. Specifically, treatment should address not only cerebral protection but also prevention of injury to other organ systems. To achieve the best outcomes, attention must be focused on optimizing blood pressure and brain tissue oxygenation, maintaining adequate cerebral perfusion pressures, and preventing seizures. In addition, maximizing good outcomes depends on proactively addressing the risk of common sequelae of brain injury, including infection, deep venous thrombosis, and inadequate nutrition. Guidelines developed for the management of severe traumatic brain injury have dramatically improved functional neurological outcomes. PMID- 19306370 TI - Traumatic brain injury: from bench to bedside to home. PMID- 19306371 TI - Cytochrome c oxidase isoform IV-2 is involved in 3-nitropropionic acid-induced toxicity in striatal astrocytes. AB - Astrocyte mitochondria play an important role for energy supply and neuronal survival in the brain. Toxic and degenerative processes are largely associated with mitochondrial dysfunction. We, therefore, investigated the effect of 3 nitropropionic acid (NPA), a mitochondrial toxin and in vitro model of Huntington's disease (HD), on mitochondrial function and viability of primary striatal astrocytes. Although NPA is known as an irreversible inhibitor of succinate dehydrogenase, we observed an increase of astrocyte ATP levels after NPA treatment. This effect could be explained by NPA-mediated alterations of cytochrome c oxidase subunit IV isoform (COX IV) expression. The up-regulation of COX isoform IV-2 caused an increased enzyme activity at the expense of elevated mitochondrial peroxide production causing increased cell death. The application of a small interfering RNA against COX IV-2 revealed the causal implication of COX isoform IV-2 in NPA-mediated elevation of oxidative stress and necrotic cell death. Thus, we propose a novel, additional mechanism of NPA-induced cell stress and death which is based on structural and functional changes of astrocyte COX and which could indirectly impair neuronal survival. PMID- 19306372 TI - Stat3 inhibition activates tumor macrophages and abrogates glioma growth in mice. AB - As the main effector-cell population of the central nervous system, microglia (MG) are considered to play an important immunoregulatory function in a number of pathological conditions such as inflammation, trauma, degenerative disease, and brain tumors. Recent studies, however, have suggested that the anti-neoplastic function of MG may be suppressed in malignant brain tumors. Considering the proposed suppressive role of signal transducers and activators of transcription 3 (Stat3) in antitumor immunity, we evaluated the role of Stat3 inhibition on MG and macrophage (MP) activation and tumor growth in a murine glioma model. N9 MG cells were exposed to GL261 glioma conditioned medium (GL261-CM) and evaluated for Stat3 activity and cytokine expression. Furthermore, the role of Stat3 inhibition on MG and MP activation was studied both in vitro and in vivo. Finally, the effect of Stat3 inhibition on tumor growth was assessed in intracranial GL261 gliomas. GL261-CM increased Stat3 activity in N9 cells in vitro and resulted in overexpression of IL-10 and IL-6, and downregulation of IL1 beta, a pro-inflammatory cytokine. Inhibition of Stat3 by CPA-7 or siRNA reversed glioma-induced cytokine expression profile in N9 cells. Furthermore, inactivation of Stat3 in intracranial GL261 tumors by siRNA resulted in MG/MP activation and tumor growth inhibition. Glioma-induced MG and MP suppression may be mediated thorough Stat3. Inhibition of Stat3 function in tumor MG/MP may result in their activation and can potentially be used as an adjunct immunotherapy approach for gliomas. PMID- 19306373 TI - Blast-related mild traumatic brain injury: mechanisms of injury and impact on clinical care. AB - Mild traumatic brain injury has been called the signature injury of the wars in Iraq and Afghanistan. In both theaters of operation, traumatic brain injury has been a significant cause of mortality and morbidity, with blast-related injury the most common cause. Improvised explosive devices have been the major cause of blast injuries. It is estimated that 10% to 20% of veterans returning from these operations have suffered a traumatic brain injury, and there is concern that blast-related injury may produce adverse long-term health affects and affect the resilience and in-theater performance of troops. Blast-related injury occurs through several mechanisms related to the nature of the blast overpressure wave itself as well as secondary and tertiary injuries. Animal studies clearly show that blast overpressure waves are transmitted to the brain and can cause changes that neuropathologically are most similar to diffuse axonal injury. One striking feature of the mild traumatic brain injury cases being seen in veterans of the wars in Iraq and Afghanistan is the high association of mild traumatic brain injury with posttraumatic stress disorder. The overlap in symptoms between the disorders has made distinguishing them clinically challenging. The high rates of mild traumatic brain injury and posttraumatic stress disorder in the current operations are of significant concern for the long-term health of US veterans with associated economic implications. PMID- 19306374 TI - Cognitive rehabilitation following traumatic brain injury: assessment to treatment. AB - Cognitive rehabilitation refers to a set of interventions that aim to improve a person's ability to perform cognitive tasks by retraining previously learned skills and teaching compensatory strategies. Cognitive rehabilitation begins with a thorough neuropsychological assessment to identify cognitive strengths and weaknesses and the degree of change in cognitive ability following a brain injury. The conclusions of the assessment are used to formulate appropriate treatment plans. Common interventions for improvements in attention, memory, and executive function, as well as the nature of comprehensive programs, which combine treatment modalities, are reviewed. Cognitive rehabilitation is effective for mild-to-severe injuries and beneficial at any time post-injury. Sufficient evidence exists supporting the efficacy and effectiveness of cognitive rehabilitation, which has become the treatment of choice for cognitive impairments and leads to improvements in cognitive and psychosocial functioning. PMID- 19306375 TI - Traumatic brain injury in the United States: an epidemiologic overview. AB - A basic description of severity and frequency is needed for planning healthcare delivery for any disease process. In the case of traumatic brain injury, severity is typically categorized into mild, moderate, and severe with information from a combination of clinical observation and self-report methodologies. Recent US civilian epidemiological findings measuring the frequency of mortality and morbidity of traumatic brain injury are presented, including demographic and etiological breakdowns of the data. Falls, motor vehicle accidents, and being struck by objects are the major etiologies of traumatic brain injury. US civilian and Army hospitalization trends are discussed and compared. Features of traumatic brain injuries from Operation Iraqi Freedom and Operation Enduring Freedom are discussed. PMID- 19306376 TI - Mild traumatic brain injury. AB - Mild traumatic brain injury accounts for 1% to 2% of emergency department visits in the United States. Up to 15% of these patients will have an acute intracranial lesion identified on head computed tomography; less than 1% of mild traumatic brain injuries will require neurosurgical intervention. Clinical research over the past decade has focused on identifying the subgroup of patients with mild traumatic brain injury with acute traumatic lesions on computed tomography and specifically those at risk for harboring a potentially catastrophic lesion. This research has been used to generate evidence-based guidelines to assist in clinical decision making. There is no evidence to support the use of plain film radiographs in the evaluation of patients with mild traumatic brain injury. The utility of brain-specific biomarkers is rapidly evolving, and a growing body of evidence supports their potential role in determining the need for neuroimaging. Clinical predictors for identifying patients with abnormal computed tomography have been established and, if used, may have a significant positive impact on traumatic brain injury-related morbidity and healthcare utilization in the United States. Patients with negative computed tomography are at almost no risk of deteriorating; however, they should be counseled regarding postconcussive symptoms and should be given appropriate written instructions and referrals at discharge. PMID- 19306377 TI - Neuroimaging of traumatic brain injury. AB - In this article, the neuroradiological evaluation of traumatic brain injury is reviewed. Different imaging strategies in the assessment of traumatic brain injury are initially discussed, and this is followed by a review of the imaging characteristics of both primary and secondary brain injuries. Computed tomography remains the modality of choice for the initial assessment of acute head injury because it is fast, widely available, and highly accurate in the detection of skull fractures and acute intracranial hemorrhage. Magnetic resonance imaging is recommended for patients with acute traumatic brain injury when the neurological findings are unexplained by computed tomography. Magnetic resonance imaging is also the modality of choice for the evaluation of subacute or chronic traumatic brain injury. Mild traumatic brain injury continues to be difficult to diagnose with current imaging technology. Advanced magnetic resonance techniques, such as diffusion-weighted imaging, magnetic resonance spectroscopy, and magnetization transfer imaging, can improve the identification of traumatic brain injury, especially in the case of mild traumatic brain injury. Further research is needed for other advanced imaging methods such as magnetic source imaging, single photon emission tomography, and positron emission tomography. PMID- 19306378 TI - Neurorehabilitation in traumatic brain injury: does it make a difference? AB - Each year, 1.4 million people in the United States are seen in a hospital for a traumatic brain injury. Those with moderate-to-severe traumatic brain injury frequently go through a course of inpatient neurorehabilitation prior to discharge back into the community. A broad overview of neurorehabilitation is presented, including the standards for admission to inpatient rehabilitation and the members and roles of the neurorehabilitation team. Common medical complications that are managed after moderate-to-severe traumatic brain injury are reviewed. The spectrum of arousal issues is summarized. The evidence regarding neurorehabilitation is then reviewed. Future studies that are underway to better understand the utility of neurorehabilitation are then discussed. PMID- 19306379 TI - Pathophysiology of traumatic brain injury. AB - Traumatic brain injury is a major source of death and disability worldwide. Significant success has been achieved in improving short-term outcomes in severe traumatic brain injury victims; however, there are still great limitations in our ability to return severe traumatic brain injury victims to high levels of functioning. Primary brain injury, due to initial injury forces, causes tissue distortion and destruction in the early postinjury period. Clinical outcomes depend in large part on mediating the bimolecular and cellular changes that occur after the initial injury. These secondary injuries from traumatic brain injury lead to alterations in cell function and propagation of injury through processes such as depolarization, excitotoxicity, disruption of calcium homeostasis, free radical generation, blood-brain barrier disruption, ischemic injury, edema formation, and intracranial hypertension. The best hope for improving outcome in traumatic brain injury patients is a better understanding of these processes and the development of therapies that can limit secondary brain injury. PMID- 19306380 TI - Differential expression of sPLA2 following spinal cord injury and a functional role for sPLA2-IIA in mediating oligodendrocyte death. AB - After the initial mechanical insult of spinal cord injury (SCI), secondary mediators propagate a massive loss of oligodendrocytes. We previously showed that following SCI both the total phospholipase activity and cytosolic PLA(2)-IV alpha protein expression increased. However, the expression of secreted isoforms of PLA(2) (sPLA(2)) and their possible roles in oligodendrocyte death following SCI remained unclear. Here we report that mRNAs extracted 15 min, 4 h, 1 day, or 1 month after cervical SCI show marked upregulation of sPLA(2)-IIA and IIE at 4 h after injury. In contrast, SCI induced down regulation of sPLA(2)-X, and no change in sPLA(2)-IB, IIC, V, and XIIA expression. At the lesion site, sPLA(2) IIA and IIE expression were localized to oligodendrocytes. Recombinant human sPLA(2)-IIA (0.01, 0.1, or 2 microM) induced a dose-dependent cytotoxicity in differentiated adult oligodendrocyte precursor cells but not primary astrocytes or Schwann cells in vitro. Most importantly, pretreatment with S3319, a sPLA(2) IIA inhibitor, before a 30 min H(2)O(2) injury (1 or 10 mM) significantly reduced oligodendrocyte cell death at 48 h. Similarly, pretreatment with S3319 before injury with IL-1 beta and TNFalpha prevented cell death and loss of oligodendrocyte processes at 72 h. Collectively, these findings suggest that sPLA(2)-IIA and IIE are increased following SCI, that increased sPLA(2)-IIA can be cytotoxic to oligodendrocytes, and that in vitro blockade of sPLA(2) can create sparing of oligodendrocytes in two distinct injury models. Therefore, sPLA(2)-IIA may be an important mediator of oligodendrocyte death and a novel target for therapeutic intervention following SCI. PMID- 19306381 TI - Neuregulin-1 beta and neuregulin-1 alpha differentially affect the migration and invasion of malignant peripheral nerve sheath tumor cells. AB - Malignant peripheral nerve sheath tumors (MPNSTs) are the most common malignancy associated with neurofibromatosis Type 1 (NF1). These Schwann cell lineage derived sarcomas aggressively invade adjacent nerve and soft tissue, frequently precluding surgical resection. Little is known regarding the mechanisms underlying this invasive behavior. We have shown that MPNSTs express neuregulin-1 (NRG-1) beta isoforms, which promote Schwann cell migration during development, and NRG-1 alpha isoforms, whose effects on Schwann cells are poorly understood. Hypothesizing that NRG-1 beta and/or NRG-1 alpha promote MPNST invasion, we found that NRG-1 beta promoted MPNST migration in a substrate-specific manner, markedly enhancing migration on laminin but not on collagen type I or fibronectin. The NRG 1 receptors erbB3 and erbB4 were present in MPNST invadopodia (processes mediating invasion), partially colocalized with focal adhesion kinase and the laminin receptor beta(1)-integrin and coimmunoprecipitated with beta(1)-integrin. NRG-1 beta stimulated human and murine MPNST cell migration and invasion in a concentration-dependent manner in three-dimensional migration assays, acting as a chemotactic factor. Both baseline and NRG-1 beta-induced migration were erbB dependent and required the action of MEK 1/2, SAPK/JNK, PI-3 kinase, Src family kinases and ROCK-I/II. In contrast, NRG-1 alpha had no effect on the migration and invasion of some MPNST lines and inhibited the migration of others. While NRG 1 beta potently and persistently activated Erk 1/2, SAPK/JNK, Akt and Src family kinases, NRG-1 alpha did not activate Akt and activated these other kinases with kinetics distinct from those evident in NRG-1 beta-stimulated cells. These findings suggest that NRG-1 beta enhances MPNST migration and that NRG-1 beta and NRG-1 alpha differentially modulate this process. PMID- 19306382 TI - Clopidogrel for preventing cardiovascular events. PMID- 19306383 TI - Physical exercise and depression. PMID- 19306384 TI - Essay: the future of traumatic brain injury. PMID- 19306385 TI - Cardiovascular safety of medication treatments for attention deficit/hyperactivity disorder. PMID- 19306386 TI - Neurobehavioral sequelae of traumatic brain injury. AB - The neurobehavioral sequelae of traumatic brain injury consist of a spectrum of somatic and neuropsychiatric symptoms. The neuropsychiatric symptoms are divided into cognitive and behavioral presentations. In the literature, these neurobehavioral sequelae have been called postconcussive symptoms, postconcussive syndrome, and postconcussive disorder; however, the authors of this review do not use this terminology because the symptoms are not restricted to patients with concussion but instead can be found in all traumatic brain injury patients of all injury severities. The development of neurobehavioral sequelae after traumatic brain injury is a multifactorial process. The patient evaluation requires a multidisciplinary approach in order to delineate physiologic dysfunction and place deficits in the context of the patient's preinjury and postinjury psychiatric status. Consequently, the evaluation of the posttraumatic brain injury patient with neurobehavioral sequelae requires a carefully structured history and physical examination with an emphasis on neurological and psychiatric function. Adjunctive evaluations must be tailored to the patient with neuroimaging, neurophysiological, and neuropsychiatric testing. Maximized outcomes may be achieved by the performance of a careful and detailed assessment that places complaints within the context of the individual. PMID- 19306387 TI - Sciatic nerve injection palsy in the child: early microsurgical treatment and long-term results. AB - Sciatic nerve injury caused by intramuscular injection in the gluteal region in the child seems as a sensory-motor palsy of the lower limb of variable degree. In preterm children or in children with severe perinatal distress, requiring intensive care, a drop foot is often missed or misdiagnosed as a malformative clubfoot or late diagnosed. Intramuscular drug injection (mainly antibiotics) during early infancy is another cause of injury. There are very few literature reports on postinjection trauma and on therapeutic indications in the child. The Authors report their experience in early microsurgical exploration of the sciatic nerve. From 1990 to 2004, we observed at different times from diagnosis 17 children with sciatic nerve palsy following intramuscular injection. Nine of them underwent nerve exploration surgery in the gluteal region (neurolysis in seven and nerve grafting in two). Conservative treatment was successful in only three cases showing early signs of recovery (at about 3 months of life). Complete recovery was observed only in five early treated cases, while late treated cases had only mild improvement after surgery. During surgery, anatomical variations predisposing to nerve injury were observed. The authors havingobserved better results and faster recovery in the early treated patients, stress the importance of a rapid therapeutic decision to avoid or limit foot deformities, sensory defects and lower limb length discrepancy due to paralysis during growth. PMID- 19306388 TI - Early stage diabetic Charcot Foot syndrome may respond to nerve decompression. AB - Diabetic Charcot Foot syndrome has been postulated to require a triggering event to initiate its puzzling inflammatory process, characterized by bony resorption, pathologic fractures, soft tissue ligamentous failure, and destruction of foot architecture. Two cases are presented where multiple lower extremity nerve decompression was performed early in the Charcot process. Resolution of clinical signs and radiographic abnormalities rapidly followed. The observation that these events were temporally concurrent suggests that nerve entrapment might reasonably be investigated as one of the postulated triggering events for the Charcot Foot in diabetes. (c) 2009 Wiley-Liss, Inc. Microsurgery 2009. PMID- 19306389 TI - Microsurgical replantation of a partial helix of the ear. AB - Microsurgical ear replantation is a significant challenge because vessel sizes are diminutive. Furthermore, as ear vessels are larger in the medial portion than in the lateral portion, microsurgical replantation of a replant including only the helix is extremely difficult. The authors replanted a partial helix of the ear using a single arterial anastomosis. As no suitable veins could be identified, medicinal leech therapy and systemic heparinization were used to achieve venous drainage. The replanted helix survived completely and the cosmetic result was excellent. Microsurgical replantation should be considered the treatment of choice in helix amputation cases, even though amount of replant is small. (c) 2009 Wiley-Liss, Inc. Microsurgery 2009. PMID- 19306390 TI - Assessment of the patency of microvascular anastomoses using microscope integrated near-infrared angiography: a preliminary study. AB - BACKGROUND: : Technical problems at the site of the anastomosis compromise an underappreciated proportion of microsurgical free tissue transfers. Intraoperative identification of technical errors may be able to prevent reexploration surgery and early flap failure. We report the first human study on a new microscope-integrated fluorescence angiography technique, which allows for intraoperative imaging of the anastomotic site. METHODS: : Fifty consecutive patients undergoing reconstructive microsurgical procedures were enrolled in the study. Intraoperative near infrared indocyanine green videoangiography (ICGA) was performed on all microsurgical anastomoses, after they had been assessed by the operating surgeon by conventional clinical patency tests. Anastomoses deemed to be occluded by the ICG-angiography were intraoperatively revised, and the result of revision was compared with angiographic findings. RESULTS: : In 11/50 (22%) of patients, where the surgeon had classified the anastomoses as patent, microangiography identified a total luminal occlusion (six) and/or significant alterations in blood flow (five), potentially predisposing toward postoperative flap failure. Intraoperative revision confirmed angiographic findings in 100% of cases, and was always associated with flap survival. The decision not to revise despite anastomotic occlusion by the intraoperative angiogram was always followed by flap loss or early reexploration. A delayed return of venous blood from the flap predisposed toward postoperative flap failure. CONCLUSIONS: : Hand-sewn anastomoses are subject to technical errors, and conventional patency tests have a low sensitivity for revealing anastomotic failure. Microscope integrated microangiography is an excellent method for identifying significant anastomotic problems, which would have otherwise gone unnoticed. The potential impact on early flap failure and reexploration surgery is considerable. (c) 2009 Wiley Liss, Inc. Microsurgery 2009. PMID- 19306393 TI - Sharing data between LSDBs and central repositories. AB - Several Locus-Specific DataBases (LSDBs) have recently been approached by larger, more general data repositories (including NCBI and UCSC) with the request to share the DNA variant data they have collected. Within the Human Genome Variation Society (HGVS) a document was generated summarizing the issues related to these requests. The document has been circulated in the HGVS/LSDB community and was discussed extensively. Here we summarize these discussions and present the concluded recommendations for LSDB data sharing with central repositories. PMID- 19306395 TI - Developmental and reproductive toxicity studies on artemisone. AB - BACKGROUND: In order to justify clinical studies in women of child-bearing age with artemisone, a new artimisinin derivative, studies to assess fertility and early embryonic development in rats, developmental toxicity in rats and rabbits, and peri-post natal development in rats were performed. METHODS AND RESULTS: In the study on fertility and early embryonic development (dose levels 0-5-20-80 mg/kg bw/day), doses inducing clinical and organ toxicity were used. Only in severe toxicity conditions, a reduction of the number of estruses, a prolonged time to insemination, decreased numbers of corpora lutea, implantation sites, and viable fetuses were found. Two developmental toxicity studies were performed in rats (dose levels 0-1-2 mg/kg bw/day) and rabbits (dose levels 0-2.5-5.0-7.5 mg/kg bw/day). It was shown that rats were about 5 times more sensitive than rabbits. In rats, artemisone induced total litter loss (late resorptions) at 2 mg/kg body weight and above with an increased incidence of a common vascular variation and retarded ossification at this dose. In rabbits, maternal toxicity, abortion and a slightly increased incidence of cardiac ventricular septal defects was observed at 7.5 mg/kg body weight. In a pre- and postnatal developmental toxicity study in rats (dose levels 0-1-2-4 mg/kg bw/day), 4 mg/kg body weight artemisone induced clinical symptoms and affected postnatal survival, body weight gain in the F1 pups, and motor activity. CONCLUSIONS: In summary, artemisone was shown to be embryo- and fetotoxic and induced cardiac ventricular septal defects and retarded ossification in dosages where total litter loss and abortions were observed. However, no effect on reproductive and developmental parameters below severe toxic dosages could be observed. PMID- 19306394 TI - Planning the human variome project: the Spain report. AB - The remarkable progress in characterizing the human genome sequence, exemplified by the Human Genome Project and the HapMap Consortium, has led to the perception that knowledge and the tools (e.g., microarrays) are sufficient for many if not most biomedical research efforts. A large amount of data from diverse studies proves this perception inaccurate at best, and at worst, an impediment for further efforts to characterize the variation in the human genome. Because variation in genotype and environment are the fundamental basis to understand phenotypic variability and heritability at the population level, identifying the range of human genetic variation is crucial to the development of personalized nutrition and medicine. The Human Variome Project (HVP; http://www.humanvariomeproject.org/) was proposed initially to systematically collect mutations that cause human disease and create a cyber infrastructure to link locus specific databases (LSDB). We report here the discussions and recommendations from the 2008 HVP planning meeting held in San Feliu de Guixols, Spain, in May 2008. PMID- 19306396 TI - Novel KCNE3 mutation reduces repolarizing potassium current and associated with long QT syndrome. AB - Long QT syndrome (LQTS) is an inherited disease involving mutations in the genes encoding a number of cardiac ion channels and a membrane adaptor protein. Among the genes that are responsible for LQTS, KCNE1 and KCNE2 are members of the KCNE family of genes, and function as ancillary subunits of Kv channels. The third KCNE gene, KCNE3, is expressed in cardiac myocytes and interacts with KCNQ1 to change the channel properties. However, KCNE3 has never been linked to LQTS. To investigate the association between KCNE3 and LQTS, we conducted a genetic screening of KCNE3 mutations and single nucleotide polymorphisms (SNPs) in 485 Japanese LQTS probands using DHPLC-WAVE system and direct sequencing. Consequently, we identified two KCNE3 missense mutations, located in the N- and C terminal domains. The functional effects of these mutations were examined by heterologous expression systems using CHO cells stably expressing KCNQ1. One mutation, p.R99lambdaH was identified in a 76-year-old woman who suffered torsades de pointes (TdP) after administration of disopyramide. Another mutation, p.T4A was identified in a 16-year-old boy and 67-year-old woman. Although the boy carried another KCNH2 mutation, he was asymptomatic. On the other hand, the woman suffered from hypokalemia-induced TdP. In a series of electrophysiological analyses, the KCNQ1(Q1)+KCNE3(E3)-R99lambdaH channel significantly reduced outward current compared to Q1+E3-WT, though the current density of the Q1+E3-T4A channel displayed no statistical significance. This is the first report of KCNE3 mutations associated with LQTS. Screening for variants in the KCNE3 gene is of clinical importance for LQTS patients. PMID- 19306397 TI - Successful stenting to coronary artery compressed by lymphoma invasion. PMID- 19306399 TI - Globus pallidus magnetization transfer ratio, T(1) and T(2) in primary biliary cirrhosis: relationship with disease stage and age. AB - PURPOSE: To determine whether the magnetization transfer ratio (MTR) of the globus pallidus (GP) in patients with primary biliary cirrhosis (PBC) correlates with age, disease stage, and fatigue, using T(1) and T(2) mapping to determine whether the mechanism of change is consistent with manganese deposition in the GP as suggested by previous reports. MATERIALS AND METHODS: In all, 30 early-stage PBC patients, four end-stage PBC patients, and 14 female controls were recruited to age-matched groups. MTR, T(1) and T(2) measurements were performed. A bilateral region of interest (ROI)-based analysis was used to calculate GP MTR, T(1), and T(2) values. These were correlated with age, disease status, and fatigue. RESULTS: MTR measurements showed a significant, negative correlation with age for controls and early-stage PBC patients, a positive correlation with T(2), and no correlation with T(1). Only GP T(2) is significantly lower in early stage PBC patients than controls, while end-stage patients demonstrated a simultaneous reduction in T(1) and MTR, consistent with GP manganese deposition. CONCLUSION: MTR measurements correlate with age in both early-stage patient and control groups, but are not associated with manganese deposition or fatigue severity: only the end-stage disease group shows changes in MTR, T(1), T(2) that are consistent with manganese deposition. PMID- 19306400 TI - Neuroimaging assessment of memory-related brain structures in a rat model of acute space-like radiation. AB - PURPOSE: To investigate the acute effects on the central nervous system (CNS) of (56)Fe radiation, a component of high-energy charged particles (HZE) in space radiation, using quantitative magnetic resonance imaging (MRI) noninvasively. MATERIALS AND METHODS: Sprague-Dawley rats were exposed to whole-brain (56)Fe (0, 1, 2, and 4 Gy). At 1 week postirradiation, MRI scans were made using T2-weighted (T2WI), diffusion-weighted (DWI), and contrast enhanced T1-(CET1) imaging. T2 relaxation time and apparent diffusion coefficient (ADC) values were obtained from memory-related brain regions of interest (ROIs). Histopathology was correlated using ex vivo tissues. RESULTS: No overt abnormalities were visualized using T2WI and DWI at 1 week postradiation. CET1 values did not differ significantly between the irradiated and control animals. Compared to 0 Gy, there were significant prolongations in T2 values and reductions in ADC after irradiation. In the absence of evident neuronal pathology, immunohistochemistry revealed astrocytic activation in 4 Gy animals. CONCLUSION: At 1 week after whole brain (56)Fe exposure, T2 and ADC values can differentiate radiosensitivity in regions critical for hippocampal-related memory. MRI may provide noninvasive assessment of the initial molecular/cellular disturbances in vivo after HZE irradiation. PMID- 19306401 TI - Correlation of iron in the hippocampus with MMSE in patients with Alzheimer's disease. AB - PURPOSE: To investigate the brain iron deposits in patients with Alzheimer's disease (AD) and healthy age-matched controls using phase imaging. MATERIALS AND METHODS: Twenty-six AD patients and 24 healthy controls were recruited. A three dimensional high-resolution, gradient-echo sequence was used to acquire phase data in the coronal plane. A high-pass filter was used to remove the phase variation caused by field inhomogeneity. The regions evaluated included the bilateral putamen, globus pallidus, and the head and body of the hippocampus. RESULTS: Significantly lower phase values in both the basal ganglion and hippocampus were revealed in the AD group compared to the normal controls (P < 0.05). The phase value in the right side of the head of the hippocampus had a moderate positive correlation with the MMSE score (r= 0.603, P = 0.000) and a negative correlation with the duration of the disease (r = -0.677, P = 0.013). Using -0.0972 radians as an optimal cutoff value, the sensitivity and specificity for differentiation between AD and normal controls reached 95.8 and 80.8%, respectively. CONCLUSION: Phase imaging proved to be a useful method for the differentiation between normal controls and AD patients. An investigation of the excessive accumulation of iron in the hippocampus may help us better understand the pathologic process and neuropsychological dysfunction of AD disease. PMID- 19306402 TI - Strain-encoded MRI to evaluate normal left ventricular function and timing of contraction at 3.0 Tesla. AB - PURPOSE: To define the reproducibility of strain-encoded (SENC) magnetic resonance imaging (MRI) for assessment of regional left ventricular myocardial strain and timing of contraction in a 3T MRI system. MATERIALS AND METHODS: The study population consisted of 16 healthy subjects. SENC measurements were performed in three short-axis (SA) slices (apical, mid, and basal) and three long axis (LA) views (two-, three-, and four-chamber) for assessment of maximal transmural systolic strain and time to peak strain. To assess the interobserver and interstudy reproducibility, analysis of SENC MRI was performed by two independent observers who were blinded to each other's results and four studies were repeated on a different day. RESULTS: Maximal longitudinal strain was highest at the apex, as was maximal circumferential strain. Peak longitudinal strain occurred earliest at the base, as did peak circumferential strain. Interclass correlation coefficient between observers and repeated studies ranged from 0.92 to 0.98 (P < 0.001 for all). CONCLUSION: The present study demonstrates the ability of SENC MRI to define regional left ventricular strain and the time sequence of regional strain. SENC MRI may represent a highly objective method for quantifying regional left ventricular function. PMID- 19306403 TI - Self-tracking of contrast kinetics for automatic triggering of contrast-enhanced whole-heart coronary magnetic resonance angiography. AB - PURPOSE: To develop a method for automatically triggering centric data acquisition during contrast-enhanced whole-heart coronary magnetic resonance angiography (MRA). MATERIALS AND METHODS: The hypothesis of this work is that the blood signal changes during contrast infusion can be estimated by obtaining a projection of the heart during inversion-recovery prepared data acquisition. A validation study was performed on seven healthy volunteers to test this hypothesis. The peak blood signal enhancement detected from the projection was then used to automatically trigger the start of central k-space data acquisition. Simulations were performed to compare the signal-to-noise ratio (SNR) of the proposed self-triggering method with the fixed delay method. Six healthy volunteers were scanned on a 3T MR system using the proposed self-triggered method to test its effectiveness on coronary artery visualization. RESULTS: Based on the validation study, the self-triggering method provides an accurate representation of the contrast enhancement. Based on the simulations, self triggering with centric ordering is expected to give a 27% higher SNR than linear ordering with a fixed imaging delay. Self-triggering was successfully used in all volunteers and showed excellent depiction of the major coronary arteries. CONCLUSION: The self-triggering method can be used to automatically determine the optimal delay time for central k-space acquisition, for each individual subject, without the need of any extra setup or user interaction. PMID- 19306404 TI - Accelerated phase-contrast MR imaging: comparison of k-t BLAST with SENSE and Doppler ultrasound for velocity and flow measurements in the aorta. AB - PURPOSE: To evaluate differences in velocity and flow measurements in the aorta between accelerated phase-contrast (PC) magnetic resonance imaging (MRI) using SENSE and k-t BLAST and in peak velocity to Doppler ultrasound. MATERIALS AND METHODS: Two-dimensional PC-MRI perpendicular to the ascending and descending aorta was performed in 11 volunteers using SENSE (R = 2) and k-t BLAST (2-, 4-, 6 , and 8-fold). Peak velocity, mean velocity, and stroke volume of the accelerated PC-MRI experiments were correlated. Peak velocities were compared to Doppler ultrasound. RESULTS: All acceleration techniques showed significant correlations for peak velocity with Doppler ultrasound. However, k-t BLAST 6 and 8 showed a significant underestimation. Strong correlations between SENSE and k-t BLAST were found for all three parameters. Significant differences in peak velocity were found between SENSE and all k-t BLAST experiments, but not for 2-fold k-t BLAST in the ascending aorta, and 2- and 4-fold k-t BLAST in the descending aorta. For mean velocity no significant differences were found. Stroke volume showed significant differences for all k-t BLAST experiments in the ascending and for 6- and 8-fold k-t BLAST in the descending aorta. CONCLUSION: Peak velocity of accelerated PC-MRI correlated with CW Doppler measurements, but high k-t BLAST acceleration factors lead to a significant underestimation. SENSE with R = 2 and 2-fold k-t BLAST are most highly correlated in phase-contrast flow measurements. PMID- 19306405 TI - The in vivo diagnosis of early-stage aortic valve sclerosis using magnetic resonance imaging in a rabbit model. AB - PURPOSE: To use magnetic resonance imaging (MRI) to identify and monitor early aortic valve sclerosis (AVS) induced by cholesterol feeding in rabbits. AVS is a highly prevalent disease process, affecting more than 25% of the population over age 65. A major obstacle to early stage medical management of AVS has been the lack of an objective noninvasive technique to identify its presence and monitor its progress. MATERIALS AND METHODS: Retrospectively gated CINE fast spoiled gradient echo (fSPGR) images of aortic valve cusps were collected at 3-month intervals starting at 6 months using a 1.5 T MR scanner interfaced with a customized surface radiofrequency coil. At 16 months a subset of animals was sacrificed and excised cusps were examined with both high frequency ultrasound (US) and histopathological techniques to validate the MRI method. RESULTS: MR and US analysis identified significant thickening of diseased AV cusps when compared to control (P < 0.05). Histopathological analysis confirmed the presence of human like AVS in diseased rabbit valves. CONCLUSION: Early AVS, exemplified by increased valve thickness, can be identified in vivo using high-resolution MRI. PMID- 19306406 TI - Influence of myocardial fibrosis and blood oxygenation on heart T2* values in thalassemia patients. AB - PURPOSE: To determine whether T2* measurements quantifying myocardial iron overload in thalassemia patients are influenced by myocardial fibrosis and blood oxygenation. MATERIALS AND METHODS: Multislice multiecho T2* was performed in 94 thalassemia patients in order to quantify myocardial iron overload. The left ventricle was automatically segmented into a 16-segment standardized heart model, and the T2* value on each segment as well as the global T2* were calculated. Delayed enhanced cardiovascular magnetic resonance (DE-CMR) images were obtained to detect myocardial fibrosis. The blood oxygenation was assessed by the noninvasive measurement of partial pressure of oxygen (pO2). RESULTS: Myocardial fibrosis was detected in 31 patients (33%). The global T2* value in patients with fibrosis was comparable with that of patients without fibrosis (P = 0.88) and T2* values in segments with fibrosis were comparable with those in segments without fibrosis (P = 0.83). The global T2* value was not correlated with the pO2 (Spearman's coefficient of correlation = 0.99). CONCLUSION: Myocardial fibrosis and blood oxygenation did not significantly affect the T2* values. These data further support the use of heart T2* as equivalent of heart iron in the clinical arena. PMID- 19306407 TI - Feasibility of quantifying the mechanical properties of lung parenchyma in a small-animal model using (1)H magnetic resonance elastography (MRE). AB - PURPOSE: To evaluate the feasibility of spatially resolving the shear modulus of lung parenchyma using conventional (1)H magnetic resonance elastography (MRE) imaging techniques in a small animal model. MATERIALS AND METHODS: A 10-cm diameter transmit-receive radiofrequency coil was modified to include a specimen stage, an MRE pneumatic drum driver, and needle system. MRE was performed on 10 female Sprague-Dawley rats using a (1)H spin-echo based MRE imaging sequence with a field of view of 7 cm and slice thickness of 5 mm. Air-filled lungs were imaged at transpulmonary inflation pressures of 5, 10, and 15 cm H(2)O while fluid filled lungs were imaged after infusion of 4 mL of normal saline. RESULTS: The average shear modulus of air-filled lungs was 0.840 +/- 0.0524 kPa, 1.07 +/- 0.114 kPa and 1.30 +/- 0.118 kPa at 5, 10, and 15 cm H(2)O, respectively. Analysis of variance indicated that these population means were statistically significantly different from one another (F-value = 26.279, P = 0.00004). The shear modulus of the fluid-filled lungs was 1.65 +/- 0.360 kPa. CONCLUSION: It is feasible to perform lung MRE in small animals using conventional MR imaging technologies. PMID- 19306408 TI - Comparison of dynamic MRI at 3.0 T and MDCT of pancreatobiliary disease: evaluation with source, MPR, CPR, and MIP images for image quality and hepatic arterial and portal venous vessel conspicuity. AB - PURPOSE: To compare contrast material-enhanced three-dimensional (3D) magnetic resonance imaging (MRI) at 3.0T and multidetector row computed tomography (MDCT) in the same patient with regard to image quality of pancreatobiliary disease and hepatic vascular conspicuity. MATERIALS AND METHODS: This study enrolled 32 patients with pancreatobiliary disease who underwent both gadolinium-enhanced 3D dynamic MRI and multiphasic CT using 16-MDCT. Data analysis of image quality was performed by two radiologists based on source images, multiplanar reconstruction (MPR), curved planar reconstruction (CPR), and maximum intensity projection (MIP) reconstruction. Determination of image quality was based on a 4-point image quality rating scale. RESULTS: The overall image quality of the MRI axial images was superior to that of the axial MDCT images. The MRI protocol yielded an average score of 3.8 points versus 3.5 for the CT imaging. No significant difference was found between 3.0T MRI and MDCT images in MPR or CPR image quality. Image quality for visualization of the distal intrahepatic segmental arteries was significantly improved using MDCT imaging. No significant difference was found between the MDCT and 3.0T MR in portal vein branch image quality. CONCLUSION: High-resolution dynamic contrast-enhanced MR imaging at 3.0T is a comprehensive technique which provides high image quality in pancreatobiliary disease. PMID- 19306409 TI - Assessment of iron distribution between liver, spleen, pancreas, bone marrow, and myocardium by means of R2 relaxometry with MRI in patients with beta-thalassemia major. AB - PURPOSE: To investigate the correlation between the degree of hepatic, splenic, pancreatic, vertebral bone marrow (VBM), and myocardial siderosis, as expressed by relaxation rate (R2 = 1/T2) values, in patients with thalassemia. MATERIALS AND METHODS: R2 relaxation rate values of liver, spleen, VBM, pancreas, and myocardium were estimated in 68 consecutive transfusion-dependent patients with beta-thalassemia major and 10 healthy controls using a respiratory triggered 16 echo Carr-Purcell-Meiboom-Gill (CPMG) spin echo sequence. RESULTS: Hepatic R2 values were significantly increased in all 68 patients; VBM, pancreatic, and myocardial R2 values were increased in 67/68, 35/47, and 47/61 patients, whereas five patients showed decreased pancreatic R2 attributed to fatty degeneration. Of the 39 nonsplenectomized patients, splenic R2 values were decreased in 30 and normal in nine patients. Hepatic R2 values correlated with splenic (r = 0.63, P < 0.001), VBM (r = 0.52, P < 0.001), but not with myocardial and pancreatic R2 values. CONCLUSION: Despite positive correlations between the degree of hepatic, splenic, and VBM siderosis, as expressed by respective R2 values, there was variability of iron distribution patterns in thalassemic patients. Unpredictable patterns of iron distribution may be seen, such as normal signal of the spleen in the presence of siderotic liver, resembling primary hemochromatosis. Fatty degeneration of the pancreas was not uncommon. PMID- 19306410 TI - Combining fine needle aspiration with brushing cytology has improved yields in diagnosing pancreatic ductal adenocarcinoma. AB - The aim of this retrospective study is to evaluate the diagnostic yields of combining fine needle aspiration (FNA) with brushing cytology (BC) in clinical work-up of pancreatic ductal adenocarcinoma. The study included a total of 97 patients who underwent both FNA and BC along with histologic/clinical follow-up (F/U). Cytologic diagnoses were categorized as negative for neoplasm (NEG), atypical/favor neoplasm (AN), and suspicious or positive for neoplasm (POS). Based on the cytologic diagnoses, the cohort was divided as follows: 23 had concordant FNA and BC diagnoses of POS/AN, all were neoplasms on F/U; 34 had disconcordant (POS/AN vs. NEG) FNA and BC diagnoses, all but 2 were neoplasms on F/U; The remaining 40 were NEG on both FNA and BC, F/U revealed that 10 were neoplasms and 30 were chronic pancreatitis. Overall, FNA rendered more true positive diagnoses than BC. However, BC but not FNA detected neoplasms in 10 patients. Most of the neoplasms identified on F/U were ductal adenocarcinoma (59 of 65). Diagnostic sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 69.2, 93.8, 95.7, 60, and 77.3% for FNA alone, 50.8, 100, 100, 50.0, and 67.0% for BC alone, and 84.6, 100, 100, 76.2, and 89.7% for combining FNA with BC. In conclusion, both EUS-guided FNA and BC are valuable modalities in the preoperative diagnosis of pancreatic ductal adenocarcinoma. When used in combination, the two modalities complement each other and achieve better diagnostic yield in pancreatic ductal adenocarcinoma than either FNA or BC alone. PMID- 19306411 TI - Primary angiosarcoma of the thyroid gland with recurrence diagnosed by fine needle aspiration: a case report. AB - Angiosarcoma of the thyroid is a rare and aggressive primary malignant tumor of the thyroid originally reported in patients from the Swiss Alpine region. Diagnosis of this tumor rests mainly on characteristic histopathological features of a malignant vascular tumor supported by immunopositivity for vascular markers e.g., CD31, Factor VIII, and CD34. Its cytological features, however, are not well-defined. We describe a case of primary angiosarcoma of the thyroid in a 48 year-old female, who presented with a rapidly enlarging neck mass associated with compressive symptoms. She had a history of hypothyroidism. The initial fine needle aspiration cytology of the neck mass was negative. She then underwent left hemithyroidectomy. Histologically, the tumor showed poorly differentiated malignant cells with eccentrically-placed nuclei, prominent nucleoli, and intracytoplasmic vacuoles admixed with mixed inflammatory cells. These showed immunopositivity for CD31 but were negative for CD34, Factor VIII, CK5/6, EMA, TTF-1, Thyroglobulin, Calcitonin, Melan A, and Calretinin. A diagnosis of poorly differentiated malignant tumor consistent with angiosarcoma was made. The patient was treated with radiation therapy but developed recurrence of the tumor. Second aspiration cytology of the recurrent tumor yielded hypocellular smears containing singularly dispersed atypical cells having eccentrically-placed nuclei with prominent macronucleoli and intracytoplasmic vacuoles within a background of inflammatory cells, consistent with recurrent angiosarcoma. Chemotherapy was started but she succumbed to the disease 7 months after diagnosis. The cytological, histopathological, immunohistochemical findings, and the clinical course are discussed. PMID- 19306412 TI - A phase 2 study with a daily regimen of the oral mTOR inhibitor RAD001 (everolimus) in patients with metastatic clear cell renal cell cancer. AB - BACKGROUND: Everolimus, an oral mammalian target of rapamycin (mTOR) inhibitor, affects tumor growth by blocking growth factor stimulation, arresting cell cycle progression, and inhibiting angiogenesis. mTOR inhibitors and agents with primarily antiangiogenic activity have been shown to have efficacy in renal cell cancer (RCC). This phase 2 study assessed the efficacy of daily oral dosing with everolimus in patients with RCC. METHODS: Patients had confirmed predominantly clear cell RCC; had received or=3 months was reported in 27 (73%) patients, and stable disease lasting >or=6 months was reported in 21 (57%) patients. Nausea (38% of patients), anorexia (38% of patients), diarrhea (31% of patients), stomatitis (31% of patients), pneumonitis (31% of patients), and rash (26% of patients) were common. Grade 3 of 4 adverse events included pneumonitis (18% of patients); transaminase elevations (10% of patients); thrombocytopenia, hyperglycemia, and alkaline phosphatase elevations (8% each of patients); and hyperlipidemia (5% of patients). CONCLUSIONS: In the current study, everolimus demonstrated encouraging antitumor activity against metastatic RCC as indicated by a PFS >or= 6 months for approximately 70% of patients. PMID- 19306413 TI - Exposure of HepG2 cells to low levels of PAH-containing extracts from contaminated soils results in unpredictable genotoxic stress responses. AB - Contaminated soil is a serious environmental problem, constituting a risk to humans and the environment. Polycyclic aromatic hydrocarbons (PAHs) are often present at contaminated sites. However, risk levels are difficult to estimate because of the complexity of contaminants present. Here, we compare cellular effects of extracts from contaminated soils collected at six industrial settings in Sweden. Chemical analysis showed that all soils contained complex mixtures of PAHs and oxy-PAHs. Western blotting and immunocytochemistry were used to investigate DNA damage signaling in HepG2 cells exposed to extracts from these soils. The effects on phosphorylated Mdm2, p53, Erk, H2AX, 53BP1, and Chk2, cell cycle regulating proteins (cyclin D1 and p21), and cell proliferation were compared. We found that most soil extracts induced phosphorylation of Mdm2 at the 2A10 epitope at low concentrations. This is in line with previous studies suggesting that this endpoint reflects readily repaired DNA-damage. However, we found concentration- and time-dependent gammaH2AX and 53BP1 responses that were sustained for 48 hr. These endpoints may reflect the presence of different types of persistent DNA-damage. High concentrations of soil extracts decreased cyclin D1 and increased p21 response, indicating cell cycle arrest. Phosphorylation of Mdm2 at Ser166, which attenuates the p53 response and is induced by many tumor promoters, was induced in a time-dependent manner and was associated with Erk phosphorylation. Taken together, the PAH extracts elicited unpredictable signaling responses that differed between samples. More polar compounds, i.e., oxy-PAHs, also contributed to the complexity. PMID- 19306414 TI - Respiratory self-gating for free-breathing abdominal phase-contrast blood flow measurements. AB - PURPOSE: To demonstrate the feasibility of using a free-breathing (FB) respiratory self-gated (RSG) approach for abdominal phase-contrast (PC) blood flow measurements. MATERIALS AND METHODS: PC-magnetic resonance imaging (MRI) flow measurements were performed within the right renal artery, common hepatic artery, and main portal vein during breath-hold (BH) and FB with both signal averaging and RSG in eight healthy volunteers. The resultant images were qualitatively scored by two independent reviewers blinded to acquisition techniques. Blood flow volume and cross-sectional vessel size measurements were compared for three techniques. RESULTS: The overall efficiency for the RSG-PC sequence was 38.9% +/- 4.7%. Images acquired with RSG effectively mitigated respiratory motion artifacts, which were clearly evident within FB signal averaged images. RSG produced similar image quality to that of BH techniques (P > 0.146) and resulted in similar vessel size measurements (P = 0.694). Flow results for both FB RSG and signal-averaged reconstructions correlated well with BH flow measurements (r = 0.97 and 0.92, P < 0.001). However, only the RSG methods demonstrated excellent absolute agreement with BH-PC flow measurements (P = 0.600), with signal-averaged methods resulting in significant overestimations. CONCLUSION: RSG methods can limit respiratory motion artifacts to reduce flow measurement inaccuracies during free-breathing PC measurements in the abdomen. PMID- 19306415 TI - Quantitative and qualitative comparison of 1.5 and 3.0 Tesla MRI in patients with chronic liver diseases. AB - PURPOSE: To compare the quantitative and qualitative image quality intra individually, at 1.5 and 3.0 Tesla (T) in patients with chronic liver diseases. MATERIALS AND METHODS: The study group included 24 consecutive patients (17 males, 7 females; mean age +/- standard deviation 56.5 +/- 11.5) who had chronic liver diseases and underwent abdominal MRI for the liver evaluation at both 1.5 and 3.0 T within a 4-month period. All MRI studies were retrospectively evaluated quantitatively and qualitatively. Quantitative analysis was performed by measuring signal to noise ratio (SNR) on various abdominal organs. Qualitative analysis was performed by two reviewers to assess image quality, artifacts, and imaging findings of chronic liver diseases. Quantitative and qualitative analyses findings were compared between 1.5 and 3.0 T using the paired Student t-test and Wilcoxon signed rank test, respectively. RESULTS: The statistically significant increase in SNRs in various abdominal tissues ranged from 1.3- to 3.5-fold at 3.0 T compared to 1.5 T. Three-dimensional gradient echo (3D-GE) sequences demonstrated significantly higher image quality at 3.0 T (P < 0.01), whereas precontrast spoiled gradient echo (SGE) sequences demonstrated significantly higher image quality at 1.5 T (P < 0.01). T2-weighted sequences did not show any significant difference in image quality between 1.5 and 3.0 T (P > 0.05). CONCLUSION: The SNRs of various abdominal tissues demonstrated significant increases at 3.0 T. The image quality of 3D-GE sequences was higher at 3.0 T, whereas the image quality of precontrast SGE sequences was higher at 1.5T. PMID- 19306416 TI - Free-breathing diffusion-weighted imaging for the assessment of inflammatory activity in Crohn's disease. AB - PURPOSE: To investigate the application of free-breathing diffusion-weighted MR imaging (DWI) to the assessment of disease activity in Crohn's disease. MATERIALS AND METHODS: Thirty-one patients with Crohn's disease were investigated using free-breathing DWI without special patient preparation or IV or intraluminal contrast agent. The bowel was divided into seven segments, and disease activity was assessed visually on DWI. For quantitative analysis, the apparent diffusion coefficient (ADC) was measured in each segment. The findings of a conventional barium study or surgery were regarded as the gold standard for evaluating the diagnostic ability of DWI to assess disease activity. RESULTS: Upon visual assessment, the sensitivity, specificity, and accuracy for the detection of disease-active segments were 86.0, 81.4, and 82.4%, respectively. In the quantitative assessment, the ADC value in the disease-active area was lower than that in disease-inactive area in small and large bowels (1.61 +/- 0.44 x 10(-3) mm(2)/s versus 2.56 +/- 0.51 x 10(-3) mm(2)/s in small bowel and 1.52 +/- 0.43 x 10(-3) mm(2)/s versus 2.31 +/- 0.59 x 10(-3) mm(2)/s in large bowel, respectively, P<0.001). CONCLUSION: Free-breathing DWI is useful in the assessment of Crohn's disease. The accuracy of DWI is high in evaluating disease activity, especially in the small bowel, and the ADC may facilitate quantitative analysis of disease activity. PMID- 19306417 TI - Platinum/taxane-based chemotherapy with or without radiation therapy favorably impacts survival outcomes in stage I uterine papillary serous carcinoma. AB - BACKGROUND: A study was undertaken to determine recurrence patterns and survival outcomes of stage I uterine papillary serous carcinoma (UPSC) patients. METHODS: A retrospective, multi-institutional study of stage I UPSC patients diagnosed from 1993 to 2006 was performed. Patients underwent comprehensive surgical staging; postoperative treatment included observation (OBS); radiotherapy alone (RT); or platinum/taxane-based chemotherapy (CT) +/- RT. RESULTS: The authors identified 142 patients with a median follow-up of 37 months (range, 7-144 months). Thirty-three patients were observed, 20 received RT alone, and 89 received CT +/- RT. Twenty-five recurrences (17.6%) were diagnosed, and 60% were extrapelvic. Chemotherapy-treated patients experienced significantly fewer recurrences than those treated without chemotherapy (P = .013). Specifically, CT +/- RT patients had a lower risk of recurrence (11.2%) compared with patients who received RT alone (25%, P = .146) or OBS (30.3%, P = .016). This effect was most pronounced in stage IB/IC (P = .007). CT- and CT + RT-treated patients experienced similar recurrence. After multivariate analysis, treatment with chemotherapy was associated with a decreased risk of recurrence (P = .047). The majority of recurrences (88%) were not salvageable. Progression-free survival (PFS) and cause-specific survival (CSS) for chemotherapy-treated patients were more favorable than for those who did not receive chemotherapy (P = .013 and .081). Five-year PFS and CSS rates were 81.5% and 87.6% in CT +/- RT, 64.1% and 59.5% in RT alone, and 64.7% and 70.2% for OBS. CONCLUSIONS: Stage I UPSC patients have significant risk for extrapelvic recurrence and poor survival. Recurrence and survival outcomes are improved in well-staged patients treated with platinum/taxane-based chemotherapy. This multi-institutional study is the largest to support systemic therapy for early stage UPSC patients. PMID- 19306418 TI - Cytodiagnosis of Ollier's disease: a case report. AB - Ollier's disease is a rare nonhereditary condition where patients present with multiple enchondromas involving usually the small bones of the hands and feet. We report the case of a 17-year-old girl who presented with multiple osteolytic lesions involving almost all phalanges of both her hands. Fine-needle aspiration cytology (FNAC) was performed from her left index finger. Smears showed mostly chondroid matrix with singly scattered cells showing binucleation and mild atypia at places. A diagnosis of enchondroma was offered in view of the cytologic and radiological findings. The dysplastic cartilage in Ollier's disease may show features similar to well differentiated chondrosarcoma. It is important not to overdiagnose such cases with atypia as low grade chondrosarcomas. Radiological appearances must be taken into account. FNAC is helpful as an outpatient diagnostic procedure when it obviates surgical intervention as most lesions tend to regress as the skeleton matures. PMID- 19306419 TI - Lymph node ratio in melanoma: A marker of variation in surgical quality? PMID- 19306420 TI - Cytomorphologic manifestations of Alzheimer's disease using brain squash smears: an autopsy study with histology-cytology correlation. AB - Alzheimer's disease (AD) is the most common form of dementia. The cardinal histopathologic features include senile plaques (SPs) and neurofibrillary tangles (NFTs), and in addition, granulovacuolar degeneration (GVD) and Hirano bodies (HBs) are frequently observed in the hippocampus. We studied hippocampal cytological features of AD, compared with controls. Hippocampal squash smears were prepared from 26 autopsy brains and stained with three different solutions, including Papanicolaou stain (Pap-s). The smears were evaluated for the aforementioned four structures and gliosis, and their frequency was compared between AD (n = 15) and control (n = 11) groups. Hippocampal smears of all AD cases revealed NFTs and gliosis. NFTs were light gray with thick flame-like structures on Pap-s. GVD was identified in the majority of AD cases and was most easily observed on Pap-s. SPs were difficult to identify and were seen only in AD cases. HBs were rarely identified as long light eosinophilic hyaline structures on Pap-s. Cytological findings of hippocampi reflect the characteristic histological features of AD with the exception of HBs, which are difficult to identify on smears. NFTs and gliosis, and SPs seem to be sensitive and specific cytologic markers in hippocampal smears for AD, respectively. PMID- 19306421 TI - Histological alterations following thyroid fine needle biopsy: a systematic review. AB - Thyroid fine-needle biopsy (FNB) is a simple, reliable, inexpensive, and generally safe diagnostic procedure in the management of thyroid nodules. However, the trauma inflicted by the needle may lead to various degrees of histological alterations, observed in histological specimens, if thyroidectomy follows. Post-FNB histological alterations of the thyroid (PFHAT) can generally be divided into acute and chronic. Hemorrhage is the most common acute and fibrosis the most common chronic PFHAT. Some of the PFHAT causes problems in histological assessment, making diagnosis difficult, even leading to misdiagnosis. In this review, we tried to collect and summarize all reported PFHAT cases and studies, aiming to make involved physicians, cytologists, and pathologists aware of the spectrum of PFHAT and to provide information to help in differential diagnosis and to avoid misdiagnoses, which could lead to unnecessary radical surgery and/or adjuvant therapy. PMID- 19306422 TI - Cytology of a recurrent nephrogenic adenoma in a renal transplant patient. PMID- 19306423 TI - Evaluation of aspiration cytology of the liver space occupying lesions by simultaneous examination of smears and cell blocks. AB - This study was undertaken to compare the efficacy of cytologic smears and histological sections from cell blocks in diagnosis of space occupying lesions (SOLs) of the liver and to classify the lesions on the basis of combined cytohistologic diagnosis. The study was conducted on 50 patients who had radiologically detected SOL/SOLs in the liver and ultrasound-guided fine needle aspiration of liver was done. In all the cases, both smears and cell blocks were made. Forty-seven cases were diagnosed as malignant and three as benign on both cytologic smears and cell blocks. Hepatocellular carcinoma was diagnosed in 8 (16%) cases and metastasis in 39 (78%) cases. The subtyping of malignancy could not be done on 11 (22%) cytologic smears and 8 (16%) cell block sections. However, on combined cytohistologic correlation, 17 (34%) out of these 19 (38%) cases could be subtyped. Sensitivity of cytologic smears and cell blocks in subtyping of malignancy was 72.3% and 82.9%, respectively. Combined cytohistologic diagnosis was found to be significantly better than isolated cytologic and cell block diagnosis (P < or = 0.05). To conclude, FNA of the SOLs of the liver is an effective procedure for diagnosing malignancy. However, cytological examination alone may fail to pinpoint the type of the tumor. Concomitant examination of cell block not only confirms the malignancy but also helps in subtyping it. PMID- 19306424 TI - Comparative analysis of immunoglobulin polymerase chain reaction and flow cytometry in fine needle aspiration biopsy differential diagnosis of non-Hodgkin B-cell lymphoid malignancies. AB - Single primer pair polymerase chain reaction (PCR) assays for the detection of clonal immunoglobulin heavy chain (IgH) gene rearrangements and immunophenotyping by flow cytometry have been proved as useful techniques in the diagnosis of lymphoid disorders in fine needle aspirates. However, a comparative analysis of both ancillary techniques in the same samples has not been previously performed. To compare the sensitivity of flow cytometry and PCR techniques, we made a wide prospective study of 77 fine needle aspiration biopsy (FNAB) samples from lymph nodes and extranodal lymphoid infiltrates. The adjunctive values of a single primer pair PCR amplification of IgH genes and of the immunophenotyping by flow cytometry were evaluated comparing their results with the final clinicopathological diagnosis of each patient supported by histological features and clinical follow up. Among the 24 B-cell non-Hodgkin lymphomas, monoclonal IgH bands were detected in 22 cases by PCR, and 21 cases were correctly considered B cell lymphoma by flow cytometry. A monoclonal IgH band was also detected in 1 of the 53 reactive lymphoid disorders. When both ancillary techniques were combined with morphological findings, 23 of the 24 B-cell lymphomas were correctly diagnosed but one reactive lymphoid disorder was also considered a B-cell lymphoma. We demonstrate a similar level of detection of B-cell lymphomas by single round PCR and flow cytometry techniques, and a strong adjunctive value when combined with morphological findings to diagnose correctly lymphoproliferative disorders by FNAB. However, we must be cautious with PCR results since false-positive cases can occur. PMID- 19306425 TI - Postradical vaginal trachelectomy follow-up by isthmic-vaginal smear cytology: a 13-year audit. AB - Radical trachelectomy is a fertility preserving alternative for early cervical cancer patients. This audit assesses the role of isthmic-vaginal smear in postoperative follow-up. A total of 94 patients were identified generating 913 smears. The final surgical margin was at the lower uterine segment in 37 cases (39.4%) and significantly correlated with the presence of lower uterine segment endometrial cells (LUSEC) in smears (P = 0.035). The most common abnormal diagnoses in the presence of LUSEC were ASC-US and AGUS seen in 14.2% and 11.9% of positive smears, respectively. The most common follow-up pattern was initial positive smears, which converted to negative (45.7% of patients), showing that reactive changes are another potential overcall pitfall. The only 2 central recurrences were successfully diagnosed by smears. This study summarizes our experience, emphasizing the role of isthmic-vaginal smears for early detection of central recurrence and highlighting the role of LUSEC and reactive changes as potential overcall pitfalls. PMID- 19306426 TI - Contrast enhanced MR angiography with parallel imaging in the early period after renal transplantation. AB - PURPOSE: To evaluate renal allograft vessels in the early period after kidney transplantation with three-dimensional (3D) contrast-enhanced MR angiography (3D CE MRA) using a parallel imaging technique. MATERIALS AND METHODS: Sixty-three consecutive patients were examined with 3D CE MRA and integrated SENSE technique (Sensitivity Encoding) 2 to 21 days after renal transplantation. MR angiography studies were analyzed for the presence of arterial stenosis. The degree of renal transplant artery stenosis was graded qualitatively as <50% = mild, 50-70% = moderate, 70-99% = severe, and occlusion. Four patients (6.3%) with moderate (n = 1) or severe (n = 3) arterial stenoses on CE MRA underwent selective intra arterial digital subtraction angiography. In two patients, selective intravenous digital subtraction angiography (DSA) was performed. RESULTS: Twenty-seven (42.9%) of the 63 patients had normal CE MR angiograms, 29 (46%) showed mild, 3 patients (4.8%) moderate, and 4 patients (6.3%) severe stenoses of the donor artery. In three patients, the severe stenosis of the graft artery was confirmed by surgery or intra-arterial DSA. One patient with suspicion of severe arterial stenosis on MRA had moderate vessel narrowing on DSA. Twelve months after kidney transplantation, serum creatinine levels were not significantly different in patients with mild and moderate stenoses from those without (P > 0.19) but significantly different from those with severe stenoses (P < 0.05). CONCLUSION: The incidence of mild and moderate vessel narrowing at the arterial anastomosis is unexpectedly high in the early period after kidney transplantation and is most likely due to surgery-related tissue edema. PMID- 19306427 TI - CE-MRA of the lower extremities using HYPR stack-of-stars. AB - PURPOSE: To investigate the properties of HYPR (HighlY constrained back PRojection) processing-the temporal fidelity and the improvements of spatial/temporal resolution-for contrast-enhanced MR angiography in a pilot study of the lower extremities in healthy volunteers. MATERIALS AND METHODS: HYPR processing with a radial three-dimensional (3D) stack-of-stars acquisition was investigated for contrast-enhanced MR angiography of the lower extremities in 15 healthy volunteers. HYPR images were compared with control images acquired using a fast, multiphase, 2D Cartesian method to verify the temporal fidelity of HYPR. HYPR protocols were developed for achieving either a high frame update rate or a minimal slice thickness by adjusting the acquisition parameters. HYPR images were compared with images obtained using 3D TRICKS, a widely used protocol in dynamic 3D MRA. RESULTS: HYPR images showed good temporal agreement with 2D control images. In comparison with TRICKS, HYPR stack-of-stars demonstrated higher spatial and temporal resolution. High radial undersampling factors for each time frame were permitted, typically approximately 50 to 100 compared with fully sampled radial imaging. CONCLUSION: In this feasibility study, HYPR processing has been demonstrated to improve the spatial or temporal resolution in peripheral CE-MRA. PMID- 19306428 TI - Gadolinium-enhanced magnetic resonance imaging for renovascular disease and nephrogenic systemic fibrosis: critical review of the literature and UK experience. AB - PURPOSE: To examine the positive reporting bias regarding the link with gadolinium (Gd) exposure and nephrogenic systemic fibrosis (NSF) in patients with renal impairment. This link has impacted strongly the international radiology safety guidelines. We believe that positive reporting bias has prevailed in the literature and that very few patients with a glomerular filtration rate (GFR) 15 29 mL/min (stage 4 chronic kidney disease [CKD]) should be regarded as high risk. MATERIALS AND METHODS: To examine this, we conducted the following steps: 1. A critical literature search on NSF. 2. An analysis of our centers magnetic resonance angiography (MRA) experience since 1999. 3. A survey of participating centers of the multicenter ASTRAL trial to assess whether any patients screened or enrolled into ASTRAL had developed NSF. RESULTS: The vast majority (90%) of NSF cases reported in the literature have occurred in patients with endstage renal disease treated with dialysis; very have had stable stage 4 or 5 (nondialysis) CKD. In all, 562 patients were followed up at our center: 30.4% were CKD4, 14.4% CKD5, 5.3% on dialysis, and 0.2% had renal transplants when imaged. No patients developed any symptoms or signs of NSF. In all, 347 patients were enrolled into ASTRAL on the basis of MRA (32% CKD4/5). One patient out of 45 centers (CKD5, received two Gd scans) developed NSF. Approximately 5 times as many patients were screened as were entered into ASTRAL. CONCLUSION: No cases of NSF were observed at our center. By extrapolation, 1/1735 patients screened for the ASTRAL trial developed NSF, giving a crude incidence rate of 0.06%. We would argue that patients with CKD4 can safely undergo Gd-MRA, albeit using a minimal dose of a macrocyclic agent and avoiding repeat doses. PMID- 19306429 TI - Enantiodifferentiation of alpha-hydroxyalkanephosphonic acids in 31P NMR with application of alpha-cyclodextrin as chiral discriminating agent. AB - Alpha-cyclodextrin was shown to be convenient chemical shift reagent for determination of the enantiomeric composition of alpha-hydroxyphosphonic acids by means of 31P NMR. The developed methodology appeared to be reliable, repetitive, easy to perform and simple for interpretation. Enantiomeric discrimination in the 31P NMR spectra for 12 of 13 studied hydroxyphosphonates was achieved, with baseline separation of resonances obtained for eight compounds. In those cases, the chemical nonequivalence values ranged from 0.069 to 0.313 ppm. The studies showed that enantioselectivity is strongly influenced by the solution pD and the optimal condition was found at pD 2 or 10 depending on the guest structure. On the basis of the ROESY spectra the complexation modes of selected hydroxyphosphonates with alpha-cyclodextrin was postulated. PMID- 19306430 TI - Characterization of chondroblastic osteosarcoma: gadolinium-enhanced versus diffusion-weighted MR imaging. AB - PURPOSE: To detect differences in magnetic resonance imaging (MRI) between chondroblastic osteosarcoma and the other types of osteosarcomas or chondrosarcomas using gadolinium-enhanced versus diffusion-weighted sequences. MATERIALS AND METHODS: Contrast-enhanced MRI and diffusion-weighted imaging (DWI) were performed in five chondroblastic osteosarcoma (CO) cases, 17 other types of osteosarcomas (OS), and 18 chondrosarcomas (CS). DWI was obtained with a single shot echo-planar imaging (EPI) sequence using a 1.5 T MR imager. The apparent diffusion coefficients (ADCs) of the minimum and maximum values were also obtained. The contrast-enhancement pattern was evaluated and minimum-maximum ADC value of CO was compared with other types of OS and CS. RESULTS: Both CO and CS showed a similar enhancement pattern; both showed septonodular and peripheral rim enhancement. The minimum ADC value of CO (1.24 +/- 0.10 x 10(-3)mm(2)/sec) was significantly higher than that of other types of OS (0.84 +/- 0.15 x 10( 3)mm(2)/sec) and was significantly lower than that of CS (1.64 +/- 0.20 x 10( 3)mm(2)/sec). In addition, the maximum ADC value of CO (2.28 +/- 0.20 x 10( 3)mm(2)/sec) was significantly higher than that of other types of OS (1.33 +/- 0.26 x 10(-3)mm(2)/sec). CONCLUSION: DWI appears to be more useful for differentiating between chondroblastic osteosarcoma and chondrosarcoma or other types of osteosarcoma than Gd-enhanced MRI. PMID- 19306431 TI - Magnetic resonance imaging and three-dimensional ultrasound of carotid atherosclerosis: mapping regional differences. AB - PURPOSE: To evaluate differences in carotid atherosclerosis measured using magnetic resonance imaging (MRI) and three-dimensional ultrasound (3DUS). MATERIALS AND METHODS: Ten subject volunteers underwent carotid 3DUS and MRI (multislice black blood fast spin echo, T1-weighted contrast, double inversion recovery, 0.5 mm in-plane resolution, 2 mm slice, 3.0 T) within 1 hour. 3DUS and MR images were manually segmented by two observers providing vessel wall and lumen contours for quantification of vessel wall volume (VWV) and generation of carotid thickness maps. RESULTS: MRI VWV (1040 +/- 210 mm(3)) and 3DUS VWV (540 +/- 110 mm(3)) were significantly different (P < 0.0001). When normalized for the estimated adventitia volume, mean MRI VWV decreased 240 +/- 50 mm(3) and was significantly different from 3DUS VWV (P < 0.001). Two-dimensional carotid maps showed qualitative evidence of regional differences in the plaque and vessel wall thickness between MR and 3DUS in all subjects. Power Doppler US confirmed that heterogeneity in the common carotid artery in all patients resulted from apparent flow disturbances, not atherosclerotic plaque. CONCLUSION: MRI and 3DUS VWV were significantly different and carotid maps showed homogeneous thickness differences and heterogeneity in specific regions of interest identified as MR flow artifacts in the common carotid artery. PMID- 19306432 TI - High-resolution fMRI with higher-order generalized series imaging and parallel imaging techniques (HGS-parallel). AB - PURPOSE: To develop a novel approach for high-resolution functional MRI (fMRI) using the conventional gradient-echo sequence. MATERIALS AND METHODS: Echo-planar imaging (EPI) techniques have generally been used for fMRI studies due to their fast imaging time. However, it is difficult for studying brain function at the submillimeter level using this sequence. In addition, EPI techniques have some drawbacks, such as Nyquist ghosts and geometric distortions in the reconstructed images, and subsequently require additional postprocessing to reduce these artifacts. One way of solving these problems is to acquire fMRI data by means of a conventional gradient-echo imaging sequence instead of EPI. To provide a fast imaging time, the proposed method combines higher-order generalized series (HGS) imaging with a parallel imaging technique which is called the HGS-parallel technique. RESULTS: The proposed HGS-parallel technique achieves a 12.8-fold acceleration in imaging time without the cost of spatial resolution. The proposed method was verified through the application of fMRI studies on normal subjects. CONCLUSION: This study suggests that the proposed method can be used for high resolution fMRI studies without the geometric distortion and the Nyquist ghost artifacts compared to EPI. PMID- 19306433 TI - Removing background phase variations in susceptibility-weighted imaging using a fast, forward-field calculation. AB - PURPOSE: To estimate magnetic field variations induced from air-tissue interface geometry and remove their effects from susceptibility-weighted imaging (SWI) data. MATERIALS AND METHODS: A Fourier transform-based field estimation method is used to calculate the field deviation arising from air-tissue interface geometry. This is accomplished by manually drawing or automatically detecting the sinuses, the mastoid cavity, and the head geometry. The difference in susceptibility, Deltachi, between brain tissue and air spaces is then calculated using a residual phase minimization approach. SWI data are corrected by subtracting the predicted phase from the original phase images. Resultant phase images are then used to perform the SWI postprocessing. RESULTS: Significant improvement in the postprocessed SWI data is demonstrated, most notably in the frontal and midbrain regions and to a lesser extent at the boundary of the brain. Specifically, there is much less dropout of signal after phase correction near air-tissue interfaces, making it possible to see vessels and structures that were often incorrectly removed by the conventional SWI postprocessing. CONCLUSION: The Fourier transform based field estimation method is a powerful 3D background phase removal method for improving SWI images, providing clearer images of the forebrain and the midbrain regions. PMID- 19306434 TI - Leptin and its receptor are expressed in the testis and in the epididymis of young and adult pigs. AB - Recent studies indicated that leptin, a 16 kDa hormone, is a regulatory signal in human and rodent male reproduction. This work was designed to investigate the expression of leptin and its receptor in testes and epididymides from immature and mature pigs. Immunolocalization revealed that leptin and its receptor were confined only in the interstitial compartment of immature testes, whereas both proteins were detected in Leydig cells and within seminiferous tubules of mature gonads. The immunostaining of epididymal tissues showed that leptin was absent in the epithelial cells of immature pigs but it was present in all the three regions of mature epididymides, although with a minor signal in the cauda. Conversely, leptin receptor was observed in all the epithelial cells of both immature and mature epididymides. Western blot analysis of tissue extracts detected a 16 kDa band for leptin and five/six isoforms, ranging from 120 to 40 kDa, for leptin receptor. In conclusion, this work has identified, for the first time, leptin and leptin receptor in the testis and in the epididymis of the pig showing a differential cell-type expression pattern of the two proteins in young and adult animals. Therefore, our findings suggest a possible involvement of leptin in endocrine or autocrine/paracrine control of porcine male reproductive structures. PMID- 19306435 TI - Effect of prolactin on the population of epithelial cells from ventral prostate of intact and cyproterone acetate-treated peripubertal rats: stereological and immunohistochemical study. AB - The interactions between steroid and nonsteroid hormones in the prostate are of special interest during the growth phase of the gland. The purpose of this work is to study the influence of prolactin (PL), with or without androgenic blockade, on epithelial cells from peripubertal rat ventral prostate. Twenty male peripubertal Sprague-Dawley rats were grouped as controls, or treated with cyproterone acetate (CA), CA plus PL (CA-PL), or PL. The total number (N total) of epithelial cells, and their labeling indices to proliferative cell nuclear antigen (LI PCNA), apoptosis (LI apoptosis) and androgen receptors (LI AR) were measured. CA and PL treatment significantly decrease the N total, but the LI PCNA was unchanged. We have observed a greater LI apoptosis in pharmacologically castrated animals without PL than in the rats with androgenic blockade with PL. The LI AR does not change with CA treatment in the ventral region, but the PL significantly increases it. Androgenic blockade and PL decrease the number of epithelial cells from the ventral prostate. These changes are not attributable to the decrease of cell proliferation, rather to the increase of epithelial apoptosis. The increase of cells expressing AR after treatment with PL might be attributed to the decrease of testosterone secretion caused by the hyperprolactinemia. PL does not modulate the size of the ventral prostate in prepubertal rats. PMID- 19306436 TI - A potential role of collagens expression in distinguishing between premalignant and malignant lesions in stomach. AB - Lack of clinical biomarkers for early gastric cancer without specific early symptoms leads to delayed diagnosis, which contributes to high mortality of gastric cancer. Here, we used oligonucleotide microarray to systematically examine differential gene expression among 33 samples from normal, premalignant, and malignant lesions in stomach. A focal adhesion pathway mainly composed of collagen genes was found to have a significantly different expression profile in gastric cancers compared to premalignant lesions. A subset of collagen genes efficiently separated malignant from premalignant tissues, and two representative genes COL11A1 and COL1A1 were validated in 42 tissue samples with quantitative reverse transcription-PCR and in situ hybridization. The data above suggest that focal adhesion pathway may have a role in the pathogenesis of gastric cancer, and the expression profile of collagen genes may be a potential biomarker to distinguish malignant from premalignant lesions in stomach. PMID- 19306437 TI - Normal right ventricular three-dimensional architecture, as assessed with diffusion tensor magnetic resonance imaging, is preserved during experimentally induced right ventricular hypertrophy. AB - The three-dimensional architecture of the right ventricular myocardium is a major determinant of function, but as yet no investigator-independent methods have been used to characterize either the normal or hypertrophied state. We aimed to assess and compare, using diffusion tensor magnetic resonance imaging, the normal architecture with the arrangement induced by chronic hypertrophy. We randomized 20 female 5 kg piglets into pulmonary trunk banding (N = 16) and sham operation (N = 4). Right ventricular hypertrophy was assessed after 8 weeks. The excised and fixed hearts were subject to diffusion tensor imaging to determine myocyte helical angles, and the presence of any reproducible tracks formed by the aggregated myocytes. All banding animals developed significant right ventricular hypertrophy, albeit that no difference was observed in terms of helical angles or myocardial pathways between the banded animals and sham group animals. Helical angles varied from approximately 70 degrees endocardially to -50 degrees epicardially. Very few tracks were circular, with helical angles approximating zero. Reproducible patterns of chains of aggregated myocytes were observed in all hearts, regardless of group. The architecture of the myocytes aggregated in the walls of the right ventricle is comparable to that found in the left ventricle in terms of endocardial and epicardial helical angles, however the right ventricle both in the normal and the hypertrophied state lacks the extensive zone of circular myocytes seen in the mid-portion of the left ventricular walls. Without such beneficial architectural remodelling, the porcine right ventricle seems unsuited structurally to sustain a permanent increase in afterload. PMID- 19306438 TI - Functional morphology of the nasal complex in the harbor porpoise (Phocoena phocoena l.). AB - Toothed whales (Odontoceti, Cetacea) are the only aquatic mammals known to echolocate, and probably all of them are able to produce click sounds and to synthesize their echoes into a three-dimensional "acoustic image" of their environment. In contrast to other mammals, toothed whales generate their vocalizations (i.e., echolocation clicks) by a pneumatically-driven process in their nasal complex. This study is dedicated to a better understanding of sound generation and emission in toothed whales based on morphological documentation and bioacoustic interpretation. We present an extensive description of the nasal morphology including the nasal muscles in the harbor porpoise (Phocoena phocoena) using macroscopical dissections, computer-assisted tomography, magnetic resonance imaging, and histological sections. In general, the morphological data presented here substantiate and extend the unified "phonic lips" hypothesis of sound generation in toothed whales suggested by Cranford et al. (J Morphol 1996;228:223 285). There are, however, some morphological peculiarities in the porpoise nasal complex which might help explain the typical polycyclic structure of the clicks emitted. We hypothesize that the tough connective tissue capsule (porpoise capsule) surrounding the sound generating apparatus is a structural prerequisite for the production of these high-frequency clicks. The topography of the deep rostral nasal air sacs (anterior nasofrontal and premaxillary sacs), narrowing the potential acoustic pathway from the phonic lips to the melon (a large fat body in front of the nasal passage), and the surrounding musculature should be crucial factors in the formation of focused narrow-banded sound beams in the harbor porpoise. PMID- 19306439 TI - Adrenal hemangioma-adenoma: an exceedingly rare adrenal collision tumor. AB - Adrenal collision tumors are rare clinical entities referring to separate coexisting adjacent tumors involving an adrenal gland with sharp demarcation between the two and without a substantial histologic admixture at the interface. We report a case of a 60-year-old female patient with an exceedingly rare adrenal hemangioma-adenoma collision tumor. To our knowledge, this is the first report of a collision tumor comprising an adrenal hemangioma and an adenoma. PMID- 19306441 TI - Diffusion-weighted magnetic resonance imaging detection of renal cancer presenting with diffuse peritoneal metastases in a patient with hemodialysis associated acquired cystic disease of the kidney. AB - A 59-year-old man was admitted with a large amount of ascites, cake-like omental thickening, and dialysis-associated acquired cystic disease of the kidney (ACDK). It was difficult to detect renal cancer, which was revealed as a primary site of peritoneal metastases by autopsy, with conventional cross-sectional imaging, such as enhanced computed tomography and T2, T1, and dynamic gadolinium-enhanced T1 weighted magnetic resonance imaging, because multiple renal cysts caused marked distortion of the renal parenchyma and the cancer had necrosis. We demonstrated the usefulness of diffusion-weighted imaging with a high b-factor to detect renal cancer presenting with peritoneal metastasis in a patient with ACDK. PMID- 19306440 TI - Extended daily access to cocaine results in distinct alterations in Homer 1b/c and NMDA receptor subunit expression within the medial prefrontal cortex. AB - Human cocaine addicts show altered function within the basal ganglia and the medial prefrontal cortex (mPFC) and altered glutamate function within these areas is postulated to be critical for cocaine addiction. The present project utilized a highly valid animal model of cocaine addiction, to test whether excessive use of cocaine alters glutamate function within these brain areas. Rats were trained to lever-press for i.v. saline vehicle or cocaine (0.25 mg/infusion) over seven 1 h daily sessions, after which, saline controls and half of cocaine self administering animals (brief access condition) received 10 more 1-h daily sessions, whereas the remaining cocaine animals received 10 additional 6-h daily sessions (extended access condition). One, 14, or 60 days after the last self administration session, animals were sacrificed. Tissue samples from the ventral tegmental area (VTA), nucleus accumbens (N.Acc) core and shell, and mPFC were analyzed by immunoblotting for expression of Homer1b/c, Homer2a/b, mGluR1, mGluR5, NR2a, and NR2b subunits of the NMDA receptor. Brief and extended access to cocaine failed to alter protein levels within the VTA, and produced transient and similar changes in N.Acc protein expression, which were more pronounced in the core subregion. In contrast, extended access to cocaine resulted in distinct and long lasting alterations of protein expression within the mPFC that included: increased levels of Homer1b/c at 1 day, NR2b at 14 days, and NR2a at 60 days, of withdrawal. These data support the notion that altered NMDA function within the mPFC may contribute, in part, to the transition to excessive uncontrolled consumption of cocaine. PMID- 19306442 TI - Multifocal intravascular papillary endothelial hyperplasia in the retroperitoneum and spine: a case report and review of the literature. AB - The authors present a unique case of intravascular papillary endothelial hyperplasia (IPEH) localized in the renal hilum, retroperitoneum, and spine in a patient with Wegener granulomatosis. IPEH rarely occurs in the abdomen or spine with few cases reported in the literature. No case has been reported of IPEH involving both the retroperitoneum and spine. In our case, MR imaging revealed enhancing masses in the right renal hilum and retroperitoneum as well as multiple focal enhancing lesions throughout the spine with lower thoracic ventral thecal sac compression. The diagnosis was established by resection of the renal hilum mass, and interval follow-up is deemed necessary for the remaining lesions because the long-term evolution of IPEH with conservative treatment alone is not well established. PMID- 19306443 TI - Sodium mapping in focal cerebral ischemia in the rat by quantitative (23)Na MRI. AB - PURPOSE: To validate (23)Na twisted projection magnetic resonance imaging (MRI) as a quantitative technique to assess local brain sodium concentration ([Na(+)](br)) during rat focal ischemia every 5.3 minutes. MATERIALS AND METHODS: The MRI protocol included an ultrashort echo-time (0.4 msec), a correction of radiofrequency (RF) inhomogeneities by B(1) mapping, and the use of 0-154 mM NaCl calibration standards. To compare MRI [Na(+)](br) values with those obtained by emission flame photometry in precision-punched brain samples of about 0.5 mm(3) size, MR images were aligned with a histological three-dimensional reconstruction of the punched brain and regions of interest (ROIs) were placed precisely over the punch voids. RESULTS: The Bland-Altman analysis of [Na(+)](br) in normal and ischemic cortex and caudate putamen of seven rats quantitated by (23)Na MRI and flame photometry yielded a mean bias and limits of agreement (at +/-1.96 SD) of 2% and 43% of average, respectively. A linear increase in [Na(+)](br) was observed between 1 and 6 hours after middle cerebral artery occlusion. CONCLUSION: (23)Na MRI provides accurate and reliable results within the whole range of [Na(+)](br) in ischemia with a temporal resolution of 5.3 minutes and precisely targeted submicroliter ROIs in selected brain structures. PMID- 19306444 TI - Diffusion tensor and tensor metrics imaging in intracranial epidermoid cysts. AB - PURPOSE: To explore the utility of diffusion tensor imaging (DTI) and diffusion tensor metrics (DTM) in characterizing the structural pathology of epidermoid cysts. DTI gives information about the tissue structure; a high fractional anisotropy (FA) indicates a highly structured orientation of the tissue, fibers, or white matter tracts. Based on the tensor rank, a set of three metrics has been described that can be used to measure the directional dependence of diffusion: linear anisotropy (CL), planar anisotropy (CP), and spherical anisotropy (CS). DTM takes into account the shape of diffusion anisotropy and hence may provide better insight into the orientation of structures than FA. MATERIALS AND METHODS: DTI was performed in three patients with epidermoid cysts. FA, directionally averaged mean diffusivity (Dav), exponential apparent diffusion coefficient (eADC), and DTM, such as CL, CP, and CS, were measured from the tumor core as well as from the normal-appearing white matter. Histopathological correlation was obtained. RESULTS: Epidermoid cysts showed high FA with Dav values similar to that of normal white matter. eADC maps did not show any restriction of diffusion. FA values were high, but not as high as that for the white matter. CP values were higher and CL values were lower than those obtained for the white matter in various regions. CONCLUSION: High CP values suggest preferential diffusion of water molecules along a two-dimensional geometry, which could be attributed to the well-structured orientation of keratin filaments and flakes within the tumor as demonstrated by histopathology. Advanced imaging modalities like DTI with DTM can provide information regarding the microstructural anatomy of the epidermoid cysts. PMID- 19306445 TI - Comparison of fMRI data from passive listening and active-response story processing tasks in children. AB - PURPOSE: To use functional MRI (fMRI) methods to visualize a network of auditory and language-processing brain regions associated with processing an aurally presented story. We compare a passive listening (PL) story paradigm to an active response (AR) version including online performance monitoring and a sparse acquisition technique. MATERIALS AND METHODS: Twenty children (ages 11-13 years) completed PL and AR story processing tasks. The PL version presented alternating 30-second blocks of stories and tones; the AR version presented story segments, comprehension questions, and 5-second tone sequences, with fMRI acquisitions between stimuli. fMRI data was analyzed using a general linear model approach and paired t-test identifying significant group activation. RESULTS: Both tasks showed activation in the primary auditory cortex, superior temporal gyrus bilaterally, and left inferior frontal gyrus (IFG). The AR task demonstrated more extensive activation, including the dorsolateral prefrontal cortex and anterior/posterior cingulate cortex. Comparison of effect size in each paradigm showed a larger effect for the AR paradigm in a left inferior frontal region-of interest (ROI). CONCLUSION: Activation patterns for story processing in children are similar in PL and AR tasks. Increases in extent and magnitude of activation in the AR task are likely associated with memory and attention resources engaged across acquisition intervals. PMID- 19306446 TI - (1)H and hyperpolarized (3)He MR imaging of mouse with LPS-induced inflammation. AB - PURPOSE: To evaluate the lipopolysaccharide (LPS) model of chronic obstructive pulmonary disease (COPD) in mouse with (1)H and hyperpolarized (HP) (3)He MR imaging. MATERIALS AND METHODS: Axial slices of the lung volume were acquired with HP (3)He and (1)H MRI at 4, 24, and 48 h after LPS exposure. A quantitative ventilation index was calculated from two HP (3)He acquisitions. A bronchoalveolar lavage (BAL) for a cell count was performed following magnetic resonance imaging (MRI). RESULTS: The LPS exposure resulted in a significant increase of cells in BAL, with maximum at 48 h. Lesions on (3)He images were characterized by ventilation defects, whereas lesions on (1)H images were hyperintense and were attributed to edema. The number of lesions was at maximum at 48 h. At this time point, and for both (3)He and (1)H MRI, the volume of the lesions was significantly higher for LPS-exposed mice compared to controls. At 4, 24, and 48 h the ventilation index from the (3)He data was significantly smaller for the LPS-exposed animals compared to controls. CONCLUSION: The time point 48 h after LPS exposure was advantageous for MRI evaluation. Functional read-out with (3)He MRI seems to be more sensitive than conventional (1)H MRI. PMID- 19306447 TI - NMR properties of human median nerve at 3 T: proton density, T1, T2, and magnetization transfer. AB - PURPOSE: To measure the proton density (PD), the T1 and T2 relaxation time, and magnetization transfer (MT) effects in human median nerve at 3 T and to compare them with the corresponding values in muscle. MATERIALS AND METHODS: Measurements of the T1 and T2 relaxation time were performed with an inversion recovery and a Carr-Purcell-Meiboom-Gill (CPMG) imaging sequence, respectively. The MT ratio was measured by acquiring two sets of 3D spoiled gradient-echo images, with and without a Gaussian saturation pulse. RESULTS: The median nerve T1 was 1410 +/- 70 msec. The T2 decay consisted of two components, with average T2 values of 26 +/- 2 msec and 96 +/- 3 msec and normalized amplitudes of 78 +/- 4% and 22 +/- 4%, respectively. The dominant component is likely to reflect myelin water and connective tissue, and the less abundant component originates possibly from intra axonal water protons. The value of proton density of MRI-visible protons in median nerve was 81 +/- 3% that of muscle. The MT ratio in median nerve (40.3 +/- 2.0%) was smaller than in muscle (45.4 +/- 0.5%). CONCLUSION: MRI-relevant properties, such as PD, T1 and T2 relaxation time, and MT ratio were measured in human median nerve at 3 T and were in many respects similar to those of muscle. PMID- 19306448 TI - High-resolution diffusion tensor imaging with inner field-of-view EPI. AB - PURPOSE: To demonstrate the applicability of inner field-of-view (FOV) echo planar imaging based on spatially two-dimensional selective radiofrequency excitations to high-resolution diffusion tensor imaging. MATERIALS AND METHODS: Diffusion tensor imaging of inner FOVs with in-plane resolutions of 0.90 x 0.90 mm(2) and 0.50 x 0.50 mm(2) was performed in the human brain and cervical spinal cord on a 3 T whole-body MR system. RESULTS: Using inner FOVs reduces geometric distortions in echo-planar imaging and allows for an improved in-plane resolution. Some of the crossings of transverse pontine fibers with the pyramidal tracts in the brainstem could be resolved, increased diffusion anisotropy and fiber orientation could be identified in cerebellar white matter, and the reduced diffusion anisotropy of spinal cord gray matter could be detected. CONCLUSION: Inner FOV echo-planar imaging may help to improve the spatial resolution and thus the accuracy of diffusion anisotropy and white matter fiber orientation measurements in the human central nervous system. PMID- 19306449 TI - Hepatic epithelioid hemangioendothelioma (HEH) at MRI using reticuloendothelial system (RES)-specific contrast agent: the "bright-bright sign" rather than the "bright-dark sign". PMID- 19306454 TI - Antitumor activity and prolonged survival by carbon-nanotube-mediated therapeutic siRNA silencing in a human lung xenograft model. AB - Carbon nanotubes are novel nanomaterials that are thought to offer potential benefits to a variety of biomedical and clinical applications. In this study, the treatment of a human lung carcinoma model in vivo using siRNA sequences leading to cytotoxicity and cell death is carried out using either cationic liposomes (DOTAP:cholesterol) or amino-functionalized multi-walled carbon nanotubes (MWNT - NH(+)(3)). Validation for the most cytotoxic siRNA sequence using a panel of human carcinoma and murine cells reveals that the proprietary siTOX sequence is human specific and can lead to significant cytotoxic activities delivered both by liposome or MWNT - NH(+)(3) in vitro. A comparative study using both types of vector indicates that only MWNT - NH(+)(3):siRNA complexes administered intratumorally can elicit delayed tumor growth and increased survival of xenograft-bearing animals. siTOX delivery via the cationic MWNT - NH(+)(3) is biologically active in vivo by triggering an apoptotic cascade, leading to extensive necrosis of the human tumor mass. This suggests that carbon-nanotube mediated delivery of siRNA by intratumoral administration leads to successful and statistically significant suppression of tumor volume, followed by a concomitant prolongation of survival of human lung tumor-bearing animals. The direct comparison between carbon nanotubes and liposomes demonstrates the potential advantages offered by carbon nanotubes for the intracellular delivery of therapeutic agents in vivo. The present work may act as the impetus for further studies to explore the therapeutic capacity of chemically functionalized carbon nanotubes to deliver siRNA directly into the cytoplasm of target cells and achieve effective therapeutic silencing in various disease indications where local delivery is feasible or desirable. PMID- 19306457 TI - Another biofuel blunder in the making?: might biofuel technologies be misused yet again for political ends? PMID- 19306458 TI - Colloidal gold and silver triangular nanoprisms. AB - It is now well-known that the size, shape, and composition of nanomaterials can dramatically affect their physical and chemical properties, and that technologies based on nanoscale materials have the potential to revolutionize fields ranging from catalysis to medicine. Among these materials, anisotropic particles are particularly interesting because the decreased symmetry of such particles often leads to new and unusual chemical and physical behavior. Within this class of particles, triangular Au and Ag nanoprisms stand out due to their structure- and environment-dependent optical features, their anisotropic surface energetics, and the emergence of reliable synthetic methods for producing them in bulk quantities with control over their edge lengths and thickness. This Review will describe a variety of solution-based methods for synthesizing Au and Ag triangular prismatic structures, and will address and discuss proposed mechanisms for their formation. PMID- 19306465 TI - Controlled growth of platinum nanoparticles on strontium titanate nanocubes by atomic layer deposition. AB - With an eye toward using surface morphology to enhance heterogeneous catalysis, Pt nanoparticles are grown by atomic layer deposition (ALD) on the surfaces of SrTiO(3) nanocubes. The size, dispersion, and chemical state of the Pt nanoparticles are controlled by the number of ALD growth cycles. The SrTiO(3) nanocubes average 60 nm on a side with {001} faces. The Pt loading increases linearly with Pt ALD cycles to a value of 1.1 x 10(-6) g cm(-2) after five cycles. Scanning electron microscopy images reveal discrete, well-dispersed Pt nanoparticles. Small- and wide-angle X-ray scattering show that the Pt nanoparticle spacing and size increase as the number of ALD cycles increases. X ray absorption spectroscopy shows a progression from platinum(II) oxide to metallic platinum and a decrease in Pt--O bonding with an increase in Pt--Pt bonding as the number of ALD cycles increases. PMID- 19306467 TI - Fetal nutritional status: diagnosis and future perspectives. PMID- 19306468 TI - Environmental rather than genetic fetal overgrowth: defining the difference and hints for diagnosis and management. PMID- 19306469 TI - Discrepant findings in a monoamniotic twin pregnancy affected by infantile myofibromatosis. PMID- 19306470 TI - Rupture of the fetal abdomen in prune belly syndrome. PMID- 19306471 TI - Medians and correction factors for biochemical and ultrasound markers in Chinese women undergoing first-trimester screening for trisomy 21. AB - OBJECTIVE: To establish normative values and distribution parameters of first trimester maternal serum free beta-human chorionic gonadotropin (beta-hCG), pregnancy-associated plasma protein-A (PAPP-A) and fetal nuchal translucency (NT) thickness in Chinese women and to examine the effects of covariates on their levels. METHODS: Maternal serum free beta-hCG, PAPP-A and fetal NT were measured in 9762 women presenting for first-trimester combined screening for Down syndrome at 11 to 14 weeks of gestation. Individuals' markers were converted to multiples of the median (MoM) using expected medians estimated by performing a weighted regression analysis. Multivariate regression analysis was performed to assess the influence of maternal weight, parity, ethnicity, chorionicity in twin pregnancies, smoking, insulin-dependent diabetes and mode of conception on individual marker MoM levels. RESULTS: Both free beta-hCG and PAPP-A median values demonstrated an exponential relationship with gestational age in days. Multivariate regression analysis indicated that free beta-hCG MoM was statistically significantly dependent on maternal weight (P < 0.0001) and chorionicity in twin pregnancy (both monochorionic and dichorionic P < 0.0001), that PAPP-A MoM was dependent on maternal weight (P < 0.0001), parity (P < 0.0001), chorionicity in twin pregnancy (both monochorionic and dichorionic P < 0.0001) and mode of conception (P = 0.002), and that fetal NT-MoM was dependent on maternal weight (P = 0.0006) and mode of conception (P = 0.012). CONCLUSION: Normative values have been generated to allow conversion of NT, free beta-hCG and PAPP-A to their MoM equivalents and correction factors have been determined to adjust for maternal and pregnancy characteristics for use in ethnic Chinese women undergoing first-trimester screening for aneuploidy. PMID- 19306472 TI - Examination of the secondary palate on stored 3D ultrasound volumes of the fetal face. AB - OBJECTIVES: To examine the use of oblique planes from stored three-dimensional (3D) ultrasound volumes of the fetal face for viewing the secondary palate at various gestational ages. METHODS: 3D ultrasound volumes of the fetal face acquired with surface rendering at 15-35 weeks' gestation in 31 fetuses with confirmed normal secondary palates were reviewed. The secondary palate was viewed in three oblique planes targeted at the uvula: the oblique axial, the oblique sagittal and the reverse face view. The detection and appearance of the secondary palate, including the soft palate, with these views at various gestational ages were compared. RESULTS: The various surfaces of the secondary palate could be viewed in all 31 fetuses in the oblique axial and the reverse face views, and in all except two fetuses in the oblique sagittal view. Both of these were < 19 weeks' gestation, at which age the uvula could not be identified clearly in this plane. CONCLUSIONS: The oblique axial, the oblique sagittal and the reverse face view targeted at the uvula allow visualization of the various aspects of the fetal secondary palate on 3D ultrasound in the second and third trimesters of pregnancy. The uvula could be used as a landmark for viewing the soft palate, but was not always easily identifiable before 19 weeks' gestation. PMID- 19306473 TI - Validation of new ultrasound parameters for quantifying pelvic floor muscle contraction. AB - OBJECTIVE: To determine the reliability and validity of new ultrasound parameters, measured in the polar coordinate system, for quantifying pelvic floor muscle action. METHODS: This was a prospective study, from January 2005 to December 2007, in 209 women with urodynamic stress incontinence, to validate new ultrasound parameters for quantifying pelvic floor muscle contraction. The examination of each patient included intravaginal digital palpation of voluntary pelvic floor muscle contractility and an ultrasound assessment of the positions of the bladder neck and anorectal junction at rest and during pelvic floor muscle contraction. The position of the bladder neck was expressed by bladder neck angle and bladder neck distance from the lower border of the pubic symphysis, and the position of the anorectal junction was expressed by the levator hiatal angle and sagittal hiatal diameter. The vector lengths of the motion of the bladder neck and anorectal junction during pelvic floor muscle contraction were calculated from the positions at rest and during pelvic floor muscle contraction by mathematical formulae. RESULTS: There was good inter- and intraobserver reliability of measurement of ultrasound parameters on stored volumes. During pelvic floor muscle contraction, elevated bladder neck distance and shortened sagittal hiatal diameter were valid parameters representing stronger pelvic floor muscle contractility, with shortened sagittal hiatal diameter having the best correlation (r = - 0.348, P < 0.001). CONCLUSION: The methods used in this study appear to be reliable for quantifying pelvic floor muscle action. The bladder neck distance with respect to the lower border of the pubic symphysis and the sagittal hiatal diameter were both valid parameters reflecting PFM contractility. PMID- 19306475 TI - Prenatal diagnosis of an aberrant right subclavian artery: four vessels arising from the aortic arch? PMID- 19306476 TI - Association between ultrasound findings and extent of trophoblastic invasion into the tubal wall in ampullary pregnancy. AB - OBJECTIVE: Predictive factors of damage to the Fallopian tube may guide the treatment of patients with tubal pregnancy. The aim of the present study was to investigate the association between the depth of trophoblastic invasion into the tubal wall, assessed on postoperative histological examination, with the findings obtained on transvaginal sonography (TVS) in women with ampullary pregnancy. METHODS: Women with ampullary pregnancy undergoing salpingectomy were enrolled into the study. Only women with a finding of either an embryo with cardiac activity or a tubal ring on TVS were included in the analysis, a total of 85 patients. Trophoblastic invasion was assessed postoperatively and was histologically classified as Stage I when limited to the tubal mucosa, Stage II when extending to the muscle layer and Stage III in the case of complete tubal wall infiltration. The association between findings on TVS and the stage of trophoblastic invasion was evaluated. RESULTS: There was a significant association between the findings on TVS and the depth of trophoblastic invasion (P < 0.001). All patients in whom an embryo with cardiac activity had been identified were found to have Stage II (17.9%) or Stage III (82.1%) invasion, whereas in those patients who showed a tubal ring on TVS, Stage I invasion was the most frequent finding (41.3%). CONCLUSIONS: In ampullary pregnancy, the finding on TVS of an embryo with cardiac activity is associated with deeper penetration of trophoblastic tissue into the tubal wall than is the finding of a tubal ring. PMID- 19306477 TI - Ultrasonographic measurement of thymus size in IUGR fetuses: a marker of the fetal immunoendocrine response to malnutrition. AB - OBJECTIVE: To test the hypothesis that intrauterine growth restriction (IUGR) is associated with decreased thymus size in the human fetus. METHODS: The thymus perimeter was measured in 60 consecutive IUGR fetuses at prenatal ultrasound examination. IUGR was defined as an abdominal circumference (AC) <5(th) centile. Sixty controls were identified by selection of the next consecutive appropriately grown fetus of similar gestational age (+/-1 week). To exclude fetal size effects, ratios between thymus perimeter and fetal biometry measurements including biparietal diameter (BPD), AC and femur length (FL), as well as estimated fetal weight (EFW) were compared between IUGR fetuses and controls. RESULTS: The proportion of fetuses with thymus perimeter <5(th) centile for gestation was significantly higher in IUGR fetuses than in controls (58/60 vs. 7/60, P < 0.0001). The mean thymus perimeter/BPD ratio (0.87 +/- 0.20 vs. 1.13 +/ 0.13, P < 0.0001), thymus perimeter/AC ratio (0.28 +/- 0.06 vs. 0.35 +/- 0.03, P < 0.0001), thymus perimeter/FL ratio (1.18 +/- 0.26 vs. 1.51 +/- 0.19, P < 0.001) and thymus perimeter/EFW ratio (0.05 +/- 0.01 vs. 0.06 +/- 0.01, P = 0.02) were significantly lower in IUGR fetuses than in controls. There was a significant positive correlation between the observed-to-expected mean for gestation thymus perimeter ratio and the enrollment-to-delivery interval (r = 0.44, P < 0.001). CONCLUSION: IUGR is associated with a disproportionately small thymus. This supports the hypothesis that thymic involution may be part of the fetal neuroendocrine response to intrauterine starvation. PMID- 19306478 TI - Use of ultrasound to distinguish between fetal hyperthyroidism and hypothyroidism on discovery of a goiter. AB - OBJECTIVES: To determine whether sonography can be used to distinguish hyperthyroidism from hypothyroidism in pregnancies with fetal goiter. METHODS: This was a retrospective study of 39 cases of fetal goiter. The majority of the mothers had Graves' disease. Fetuses were scanned for the existence of a hypertrophic thyroid gland (goiter) beginning at 22 gestational weeks. Once a goiter was diagnosed, different echographic features were analyzed and the effect of chosen treatment on fetal thyroid development was monitored. RESULTS: On color Doppler, 68.8% of hypothyroid goiters had a peripheral vascular pattern vs. 20% in cases of fetal hyperthyroidism (P = 0.0574). No hypothyroid goiter presented central vascularization whereas half the hyperthyroid goiters did (P = 0.0013). Fetal tachycardia was a good indicator of hyperthyroidism (57.1% v.s 6.3%; P = 0.0055). Delayed bone maturation was seen in hypothyroid goiters (46.9% vs. 0%; P = 0.0307), while advanced bone maturity was specific to hyperthyroid goiters (85.7% vs. 0%; P < 0.0001). Lastly, an increase in fetal movement was observed in cases of fetal hypothyroidism (43.8% vs. 0%; P = 0.0364). CONCLUSION: Based on the color Doppler pattern of goiter, fetal heart rate, bone maturation and fetal mobility, we established an ultrasound score to predict fetal thyroid function in cases of fetal goiter. PMID- 19306479 TI - Protein-free ligand screening: simplification of chiral chromatographic development via novel adaptation of NMR screening methodologies. AB - We demonstrate here a promising NMR method that provides evidence for chiral compound selector interaction as a first-pass screening method. A novel adaptation of commonly used protein-based screening technologies, this approach relies upon ligand-to-stationary phase interaction wherein the stationary phase is tethered to sepharose beads. At only minutes per experiment, this methodology significantly reduces the time required for chiral separation methodology development and complements currently available chromatographic purity technologies. PMID- 19306480 TI - Cardioprotective potential of myricetin in isoproterenol-induced myocardial infarction in Wistar rats. AB - The study aimed to evaluate the protective role of myricetin obtained from Vitis vinifera (Vitaceae) on heart rate, electrocardiographic (ECG) patterns, vascular reactivity to catecholamines, cardiac marker enzymes, antioxidant enzymes together with morphological and histopathological changes in isoproterenol (ISO) induced myocardial infarction (MI) in male Wistar rats. Rats treated with isoproterenol (85 mg/kg, administered subcutaneously twice at an interval of 24 h) showed a significant increase in heart rate and ST elevation in ECG, and a significant increase in the levels of cardiac marker enzymes - lactate dehydrogenase (LDH), creatine kinase (CK) and aspartate aminotransferase (AST) in serum. Isoproterenol significantly reduced superoxide dismutase (SOD) and catalase (CAT) activity and increased vascular reactivity to various catecholamines. Pretreatment with myricetin (100 mg/kg, p.o. and 300 mg/kg, p.o.) for a period of 21 days significantly inhibited the effects of ISO on heart rate, levels of LDH, CK, AST, SOD, CAT, vascular reactivity changes and ECG patterns. Treatment with myricetin (100 mg/kg and 300 mg/kg) alone did not alter any of the parameters compared with vehicle treated Wistar rats. Myricetin treated animals showed a lesser degree of cellular infiltration in histopathological studies. Thus, myricetin (100 mg/kg and 300 mg/kg) ameliorates the cardiotoxic effects of isoproterenol and may be of value in the treatment of MI. PMID- 19306481 TI - Unequivocal assignments of flavonoids from Tephrosia sp. (Fabaceae). AB - (1)H and (13)C NMR chemical shifts of praecansone B, pongaflavone and dehydrorotenone isolated from Tephrosia egregia Sandw and obovatin from T. toxicaria Pers. were unambiguously assigned by 1D and 2D NMR experiments including (1)H, (1)H COSY, gHMQC and gHMBC, allowing the correction of literature assignments. PMID- 19306482 TI - Is there a doctor in the house? (There could be another one; it depends on you!). PMID- 19306483 TI - ACP Journal Club. What does it take to put an ugly fact through the heart of a beautiful hypothesis? PMID- 19306484 TI - [Adrenal gland tumor biopsy]. PMID- 19306485 TI - ACP Journal Club. Review: Perioperative beta-blockers provide no clear benefit in patients having noncardiac surgery. PMID- 19306486 TI - [Quality requirements for wording of obituaries]. PMID- 19306487 TI - ACP Journal Club. Review: Long-acting beta-agonists do not increase risk for asthma-related hospitalization in patients taking inhaled corticosteroids. PMID- 19306488 TI - ACP Journal Club. Review: Inhaled corticosteroids do not reduce mortality but increase pneumonia in chronic obstructive pulmonary disease. PMID- 19306489 TI - ACP Journal Club. Review: Enhanced oral hygiene prevents respiratory infection in older persons in hospitals and nursing homes. PMID- 19306490 TI - ACP Journal Club. Intensive and standard glucose control did not differ for major CV events or death in poorly controlled type 2 diabetes. PMID- 19306491 TI - ACP Journal Club. Combined therapy with folic acid and vitamins B6 and B12 did not affect cancer risk in women at high risk for CV disease. PMID- 19306492 TI - ACP Journal Club. Selenium and vitamin E, alone or together, did not prevent prostate cancer. PMID- 19306493 TI - ACP Journal Club. Vitamin E and vitamin C, alone or together, did not prevent prostate or total cancer in men. PMID- 19306494 TI - ACP Journal Club. Prednisolone, but not valacyclovir, reduced time to complete recovery of facial-nerve function in Bell palsy. PMID- 19306495 TI - ACP Journal Club. Review: Combinations of clinical findings had moderate sensitivity and specificity for diagnosing the irritable bowel syndrome. PMID- 19306496 TI - ACP Journal Club. Reynolds Risk Score for men predicted cardiovascular events more accurately than traditional risk factors only. PMID- 19306497 TI - ACP Journal Club. Simplification of the revised Geneva score did not decrease accuracy for diagnosis of pulmonary embolism. PMID- 19306498 TI - Gene silencing below the immune radar. AB - In vertebrates, the detection of viral nucleic acids is the first step toward innate and subsequent adaptive antiviral immune responses. A sophisticated,protein receptor-based sensor system has evolved to recognize viral nucleic acids and to trigger a variety of antiviral defense mechanisms. The more we learn about this elaborate sensor system, the more it becomes evident how difficult it is to introduce exogenous nucleic acids such as siRNA into cells without triggering antiviral immunoreceptors. In this issue of the JCI, Judge and colleagues provide evidence that siRNA can be designed and delivered in a way that allows specific and successful silencing of target genes in tumor cells in vivo, leading to tumor cell death and prolonged survival of tumor-bearing mice in the absence of immune activation. This study represents a major technological advance, setting new standards for well-controlled siRNA applications in vivo, and has the potential to guide clinical development toward siRNA therapeutics with well-defined and selective gene-silencing activities. PMID- 19306499 TI - Protein degradation in Parkinson disease revisited: it's complex. AB - Mutations in the genes PTEN-induced putative kinase 1 (PINK1), PARKIN,and DJ-1 cause autosomal recessive forms of Parkinson disease (PD), and the Pink1/Parkin pathway regulates mitochondrial integrity and function.An important question is whether the proteins encoded by these genes function to regulate activities of other cellular compartments. A study in mice,reported by Xiong et al. in this issue of the JCI, demonstrates that Pink1,Parkin, and DJ-1 can form a complex in the cytoplasm, with Pink1 and DJ-1 promoting the E3 ubiquitin ligase activity of Parkin to degrade substrates via the proteasome. This protein complex in the cytosol may or may not be related to the role of these proteins in regulating mitochondrial function or oxidative stress in vivo. PMID- 19306500 TI - Shock the heat shock network. AB - The targeting of tumors is made possible through establishing protein signatures specific for each cancer type. The recent recognition of the higher expression levels of HSP90 and its accumulation in tumor cell mitochondria has made the HSP90 network a feasible target for neutralization. HSP90 antagonizes the mitochondrial permeability transition,blocking cytochrome c release and apoptosis. In this issue of the JCI, Kang et al. report the synthesis of Gamitrinibs, which target mitochondrially localized HSP90, specifically killing human cancer cell lines, and provide a fresh approach for cancer treatment. PMID- 19306501 TI - NO more muscle fatigue. AB - NOS is a key enzyme in the production of NO, a molecule that directly regulates vasorelaxation and blood supply. Diverse forms of muscle disease have been clinically associated with unusual fatigue after exercise. The localization of neuronal NOS (nNOS) at the plasma membrane of muscle has recently been shown to prevent muscle fatigue after exercise. In this issue of the JCI, Lai et al. show that dystrophin--the structural protein missing in individuals with Duchenne muscular dystrophy--anchors nNOS to the sarcolemma through a direct interaction with dystrophin spectrin-like repeats 16 and 17 (see the related article, doi:10.1172/JCI36612). Furthermore, in another recently reported study of mouse models of muscular dystrophy, phosphodiesterase 5A inhibitors were used to treat the downstream ischemia that is associated with nNOS mislocalization. Collectively, these findings significantly advance our understanding of exercise induced muscle fatigue and its role in muscle disease. PMID- 19306502 TI - PD-1 tempers Tregs in chronic HCV infection. AB - Adaptive T cell responses are critical for controlling infections with viruses such as HIV, HBV, and HCV. However, these responses must be carefully regulated because overly vigorous T cell activation can lead to excessive host tissue damage. T cell expression of the inhibitory receptor programmed death-1 (PD-1) and inhibition of effector T cells (Teffs) by CD4+Foxp3+Tregs are among the many described mechanisms for achieving a balanced immune response. Although the signals that contribute to Teff function are well understood, less is known about the signals controlling Tregs. In this issue of the JCI, Franceschini et al. extend our understanding of how Tregs are modulated during chronic HCV infection by demonstrating that Treg proliferation is inhibited by PD-1 and that this inhibition is mediated by a potentially novel mechanism involving the prevention of IL-2-driven STAT-5phosphorylation. PMID- 19306503 TI - Drug coupons don't always save money. PMID- 19306504 TI - [Metha-analysis of results of stress-tests application in patients before the operations on peripheral vessels and abdominal aorta]. AB - Results of meta-analysis of 90 publications concerning cardiac stress-tests carried out to patients before operations on vessels and abdominal aorta are presented; 11,778 patients were included in the system review. On the basis of literary data advantages and disadvantages of diverse diagnostic tests and contra indications to them were analyzed. Findings prove that pharmacological stress tests have the highest sensitivity and specificity. Out of various diagnostic tests--treadmill test with ECG control, Holter monitoring, radioisotope imaging with thallium-201, and dipyridamole, and stress-echocardiography with dobutamine- the last one is the most informative in predective value of positive result, relative risk of possibility development of negative cardiac event (myocardial infarction or patient's death in consequence of myocardial infarction), likelihood ratio, sensitivity and specificity. Furthermore, during stress echocardiography with dobutamine examination of myocardial contractility and evaluation of ejection fraction are available what makes it a method of choice in cardiac risk stratification in patients before reconstructive vascular surgery. PMID- 19306505 TI - [Experience in treatment of postoperative ventral hernias]. AB - Treatment results in 193 patients with medium and large postoperative ventral hernias were analyzed. All patients were divided into 2 groups. 66 patients operated according to autoplastic methods formed the control group. 127 patients undergoing abdominal wall grafting with use of cellular polypropylene implant formed the main group. Sublay method was used in 48 patients, onlay--in 38 patients, Rives-Stoppa method--in 41 patients. Frequency of local wound postoperative complications and hernia relapses were taken into account for comparative evaluation of surgical treatment results. Hernia relapses developed in 56 patients from the control group and in 115 patients from the main group. Basing on the authors' data autoplastic methods are accompanied by local wound complications (infiltration, suppuration, seroma, haematoma) in 7.9% of observations, alloplastic methods--in 22.8% (chi2 = 6.36, p<0.05). In the control group hernia relapse developed in 18 patients (32.1%), in the main group--in 6 patients (5.2%) (chi2 = 22.63, p<0.001). Hernia relapse was revealed when using onlay method in 8.6% of observations, Rives-Stoppa--in 5.4%, sublay--in 2.3% of observations (chi2 = 1.53, p>0.05). Authors consider that it is necessary to use diverse alloplasic methods treating median postoperative hernias of medium and big sizes. Preference should be given to sublay method. PMID- 19306506 TI - [Laser Doppler flowmetry in burn treatment]. AB - It is offered to use laser Doppler fluometry indices at the acute stage of burn disease for diagnostics of lesion depth, as a criterion for degree of shock severity and its ending, for evaluation of the surgical treatment influence on clinical course of burn disease, and also at different stages of wound closing with complex temporary pedicle flap. Methods and criterions of microcirculation study during the period of burn shock, and also in burn re-convalescents are defined for evaluation of conservative treatment efficacy and state of microcirculation in forming scars. Application of laser Doppler fluometry contributes to reduction of surgical operations number, duration of stay in the hospital, improvement of aesthetic treatment results. Laser Doppler fluometry indices can serve as a criterion of conservative therapy efficacy, contribute to timely correction of rehabilitation plan. PMID- 19306507 TI - Label-free molecular imaging of living cells. AB - Optical signals based on Raman scattering, coherent anti-Stokes Raman scattering (CARS), and harmonic generation can be used to image biological molecules in living cells without labeling. Both Raman scattering and CARS signals can be used to detect frequencies of molecular vibrations and to obtain the molecular distributions in samples. Second-harmonic optical signals can also be generated in structured arrays of noncentrosymmetric molecules and can be used to detect structured aggregates of proteins, such as, collagen, myosin and tubulin. Since labeling techniques using chemical and biological reactions may cause undesirable changes in the sample, label-free molecular imaging techniques are essential for observation of living samples. PMID- 19306508 TI - The GAVI Financing Task Force: one model of partner collaboration. AB - The Global Alliance for Vaccines and Immunization, now called the GAVI Alliance, was launched in 2000 as a coalition of partners, including countries, international organizations, bilateral donors, the vaccine production industry, and nongovernmental organizations; most activities were to be implemented through these partner organizations. Four task forces were established at the outset to define issues relevant to GAVI Alliance goals and to recommend actions. This paper describes the innovations and outputs of the Financing Task Force (FTF), which worked in three areas: country support to sustainably finance vaccines and immunization programs in the context of introducing new vaccines; vaccine supply and demand issues as they impact vaccine choice, production costs and price/dose; innovative financing mechanisms for vaccines and immunization programs through, for example, capital markets. This analysis particularly focuses on the FTF's work on financial sustainability. Through its partnership, the FIF was able to leverage organizational change in its participating organizations, in the countries supported by the GAVI Alliance, and in the policies of GAVI itself. These achievements, along with areas where the desired outcome was not achieved, are summarized with lessons that may be useful to other multi-partner health alliances. PMID- 19306509 TI - Advanced head and neck cancer: is there a role for fluorodeoxyglucose PET/computed tomography? PMID- 19306510 TI - Comparison of 2D and 3D performance for FDG PET with different acquisition times in oncological patients. AB - OBJECTIVE: Collecting positron emission tomography data in three-dimensional (3D) mode may potentially allow reduction of the tracer dose and/or the acquisition time without compromising image quality thereby making the procedure more patient friendly and cost effective. The objective of our study was to compare VUE Point iterative reconstruction algorithm in positron emission tomography data obtained in 3D and two-dimensional (2D) mode in routine clinical diagnostic setting in oncological patients. METHODS: Standard whole-body imaging (33 patients) was followed by rescanning of the target region in 2D and 3D mode. The ListMode data were histogrammed to 4, 3 and 2 min frames. Visual and semiquantitative analyses were performed. Effects of tumour volume and body mass index on tissue visualisation were evaluated. RESULTS: Visual image quality in 3D mode was superior to 2D. Maximum and mean standardised uptake values of tumours did not differ between 2D and 3D. Mean background standardised uptake values were significantly lower in 3D compared with 2D. 3D tumour to background ratios were higher than 2D in small lesions and obese patients (body mass index > or = 30) with all acquisition times. CONCLUSION: The new reconstruction method produces images of similar or better quality with 3D compared with 2D mode. The difference is pronounced in small lesions and in obese patients. According to our study, 3D is the preferred acquisition mode for routine whole-body evaluation of oncological patients. PMID- 19306511 TI - Dual time point FDG PET for evaluation of malignant pleural mesothelioma. AB - OBJECTIVE: To evaluate whether 2-deoxy-2-18F-fluoro-D-glucose (FDG) positron emission tomography (PET) is more useful in differentiating malignant from benign pleural lesions, and whether delayed FDG PET imaging can improve the diagnostic performance in patients with suspicion of malignant pleural mesothelioma (MPM). METHODS: Thirty-three patients who were suspected of having MPM were examined with FDG PET. PET imaging (whole body) was performed at 60 min (early) post-FDG injection and repeated at 120 min (delayed) after injection only in the thoracic region. We evaluated the FDG uptake visually and semiquantitatively. The semiquantitative analysis using the standardized uptake value (SUV) was determined for both early and delayed images (SUVearly and SUVdelayed, respectively). RESULTS: The final diagnosis was MPM in 17 patients and benign pleural disease in 16 patients. The sensitivity, specificity, and accuracy to detect MPM on both early and delayed PET were all 88%. The mean value of SUVdelayed in MPM was significantly higher than that of SUVearly (P < 0.001). The mean values of SUVearly and SUVdelayed in MPM were significantly higher than the corresponding values in benign pleural disease (P < 0.01, respectively). CONCLUSION: FDG PET seems to be a useful imaging modality for differential diagnosis of MPM. In addition, the diagnostic performance on delayed PET was the same as that on early PET, although SUVdelayed was significantly higher than SUVearly in patients with MPM. PMID- 19306512 TI - Preliminary study of carbon-11 methionine PET in the evaluation of early response to therapy in advanced breast cancer. AB - OBJECTIVE: Breast cancer is one of the principal oncological challenges in the Western world. Currently, there are only a few reliable predictive methods for monitoring treatment. We investigated the ability of carbon-11 methionine ("11C MET) positron emission tomography (PET) to evaluate early response to therapy in advanced breast cancer. METHODS: Thirteen patients with metastases in the lungs/pleura, lymph nodes, soft tissue, or bones entered a MET PET study both before and after the first cycle of polychemotherapy (n=4), or after the first month of therapy with hormones (n=5), or low dose weekly cytostatics (n=3). One patient underwent three PET studies: before hormonal therapy, after 1 month of hormonal therapy, and after the first cycle of polychemotherapy (total, 27 studies). MET accumulation in the metastatic sites was measured as standardized uptake values (SUVs), and the pretreatment and post-treatment SUVs were compared with each other and the clinical follow-up data. RESULTS: A total of 26 different metastatic sites were investigated in 13 patients. All metastases were visible by MET PET except one superficially spreading local skin recurrence, probably because of respiratory movements. Five new metastatic sites were detected. After therapy the SUVs decreased significantly (30-54%; P < 0.05) in all six responding metastatic sites, whereas the SUVs of nonresponding metastases decreased somewhat (11-130/%; n=4), remained stable (+/- 8%; n=10), or increased (13-23%; n=4) (P=NS). The SUVs of two nonresponding metastatic sites decreased clearly. Physiological MET uptake in the salivary glands, the myocardium, and the bone marrow did not disturb the image interpretation. CONCLUSION: MET PET may be useful in assessing the early response to therapy in advanced breast cancer. PMID- 19306513 TI - Clinical value of FDG-PET/CT for the diagnosis of human immunodeficiency virus associated fever of unknown origin: a retrospective study. AB - AIM: The aim of this study was to evaluate retrospectively the usefulness of [18F]fluorodeoxyglucose (FDG)-PET/ computed tomography (CT) in patients affected by human immunodeficiency virus and suffering from fever of unknown origin (HIV associated FUO). MATERIAL AND METHODS: Ten patients (six males, four females, age 24-48 years) suffering from HIV-associated FUO were studied by FDG-PET/CT. Final diagnosis was established either by microbiological or histopathological analysis or by a more than 6-month follow-up. FDG-PET/CT was regarded as 'helpful for diagnosis' when the abnormal uptake pointed to the organ or location where the cause of fever was thereafter identified. RESULTS: Nine out of 10 FDG-PET/CT were abnormal and the cause of fever was further demonstrated by other diagnostic procedures. An infectious process (tuberculosis) was diagnosed in six patients and a neoplasm in three (two lymphomas, one Kaposi's sarcoma). FDG-PET/CT directly suggested sites for biopsy in six patients (tuberculous lymphadenitis and neoplasm). The only patient with normal FDG-PET/CT suffered from drug-induced fever. CONCLUSION: FDG-PET/CT is a valuable tool in patients with HIV-associated FUO. FDG-PET/CT was categorized as 'helpful for diagnosis' in nine out of the 10 patients we studied. Adding the CT anatomical landmarks to the PET findings allowed an accurate and easy localization of the sites to be punctured in the six patients in whom histopathological diagnosis was needed. PMID- 19306514 TI - Combining chronic kidney disease with 201thallium/123iodine beta methyliodophenyl pentadecanoic acid dual myocardial single-photon emission computed tomography findings is useful for the evaluation of cardiac event risk. AB - OBJECTIVE: Chronic kidney disease is a noteworthy pathophysiology as a risk factor of cardiovascular disease. We investigated the usefulness of combining glomerular filtration rate and 201thallium(201TI)/123iodine-beta-methyliodophenyl pentadecanoic acid (123I-BMIPP) dual myocardial scintigraphic findings for predicting cardiac events. METHODS: Seventy-five patients suspected of coronary artery disease underwent 201TI/123I-BMIPP dual myocardial scintigraphy. Clinical and nuclear variables were included in the multivariate analysis for predicting hard events (cardiac death and nonfatal myocardial infarction) and soft events (unstable angina, heart failure, and coronary revascularization). Glomerular filtration rate was estimated by the modification of diet in renal disease formula. Kaplan-Meier analysis was performed to investigate the incremental prognostic value of glomerular filtration rate. RESULTS: During the mean follow up period of 425 days, eight patients had hard events and 20 patients had soft events. Multivariate analysis revealed that glomerular filtration rate and the sum of total defect score in 123I-BMIPP image were independent predictors of total cardiac events, whereas sex, diabetes, glomerular filtration rate, and the number of abnormal segments in 201TI image were those of hard events. Kaplan Meier analysis revealed that greater risk stratification was achieved by adding a glomerular filtration rate of lesser than 60 ml/min/1.73 m2 to the sum of the total defect score > or = 5 in the 123I-BMIPP image. Greater risk stratification for hard events was also achieved by adding a glomerular filtration rate of lesser than 30 ml/min/1.73 m2 to the number of abnormal segments > or = 2 in 201TI image. CONCLUSION: Better risk stratification can be achieved by adding glomerular filtration rate to 201TI/123I-BMIPP dual myocardial scintigraphic findings. PMID- 19306515 TI - Myocardial perfusion gated single-photon emission computed tomography in patients with left bundle branch block: comparison between the end-diastolic images and the ungated images. AB - OBJECTIVES: Septal perfusion abnormalities are frequently observed in patients with left bundle branch block (LBBB). The aim of this study was to compare myocardial perfusion imaging obtained from ungated and diastolic thallium gated single-photon emission computed tomography (SPECT) images in patients with LBBB. METHODS: Stress/rest SPECT was performed in 70 patients with LBBB [38 with coronary artery disease (CAD) (G1), 32 without (G2)] and 19 control participants (G3). Diastolic images were obtained as the sum of four diastolic bins. Perfusion was assessed by summed stress, rest, and difference scores for both diastolic and ungated images. RESULTS: In G1, there was no difference between diastolic and ungated perfusion scores. In G2, summed stress score and summed rest score were increased in diastolic versus ungated imaging, and perfusion defect extent was increased on diastolic versus ungated images at stress (diastole: 6.2 +/- 9.9% vs. ungated: 5.1 +/- 9.70/%, P = 0.01) and rest (diastole: 4.3 +/- 9.3% vs. ungated: 2.8 +/- 9.50%, P = 0.0014), with an increased extent of reversible defect (diastole: 3.4 4.7% vs. ungated: 2.3 3.7%, P = 0.01). In G2, diastolic images failed to correct septal perfusion abnormalities. The number of normal SPECT did not increase with diastolic versus ungated imaging (24 and 180% in G1, 66 and 53% in G2, respectively). Otherwise, a significant association between left ventricular dysfunction and CAD was found, stress ejection fraction being decreased in 20 patients (53%) in G1 and seven patients (22%) in G2 (CHI2 = 6.93, P < 0.01). CONCLUSION: In patients with LBBB, diastolic imaging did not provide additional information to ungated perfusion imaging, but left ventricular dysfunction was associated with CAD. PMID- 19306516 TI - One-year follow-up of myocardial perfusion and function evaluated by gated SPECT MIBI in patients with earlier myocardial infarction and chronic total occlusion. AB - OBJECTIVE: Optimal treatment for chronic total occlusion (CTO) in the infarct related coronary artery is not clear. Our aim was to assess myocardial perfusion, left ventricular ejection fraction (EF), and left ventricular size using gated single-photon emission computed tomography (SPECT) myocardial perfusion imaging with 99mTc-methoxy-isobutyl-isonitrile in patients with CTO before and 1 year after recanalization. METHODS: Thirty patients with earlier myocardial infarction and at least one CTO underwent percutaneous coronary intervention (PCI) as well as nitrate-enhanced gated SPECT myocardial perfusion and dobutamine stress echocardiography before and 11 +/- 1 months after recanalization. They were divided into three groups based on the outcome of the follow-up angiography: (i) successful recanalization with no evidence of in-stent restenosis (n=13); (ii) successful recanalization with in-stent restenosis (n=7) and (iii) unsuccessful recanalization (n=10). RESULTS: Overall success of recanalization for CTO was 74%. In group 1, myocardial viability was preserved in 11 of 13 (85%) patients at baseline. Gated SPECT at 1 year showed a significant decrease in perfusion abnormalities (29 +/- 12 to 23 +/- 14%, P < 0.05) and left ventricular end diastolic volume (EDV) (168 +/- 47 to 151 +/- 47 ml, P < 0.05). Improvement in EF (51 +/- 11 to 54 +/- 13%, P > 0.05) and reduction in left ventricular end systolic volume (ESV) (84 +/- 37 to 77 +/- 40 ml, P > 0.05) did not reach the level of significance. Myocardial viability was preserved in only two of seven patients (28%) in group 2. Neither mean perfusion abnormalities (37 +/- 24 to 35 +/- 22%, P > 0.05) nor global left ventricular parameters (EF 41 +/- 15 vs. 42 +/ 19%, EDV 298 +/- 33 vs. 299 +/- 57 mL, ESV 197 +/- 12 vs. 195 +/- 32 ml; P > 0.05) changed at the follow-up. In group 3, myocardial viability was preserved in seven of 10 patients (70%) at baseline, but no significant changes in perfusion (40 +/- 18 vs. 41 +/- 19%, P > 0.05) and left ventricular function (EF 42 +/- 17 vs. 44 +/- 14%, EDV 228 +/- 101 vs. 227 +/- 81 ml, ESV 143 +/- 87 vs. 146 +/- 8 ml; P > 0.05) were seen at the follow-up. CONCLUSION: Myocardial perfusion and EDV may significantly improve 1 year after PCI provided recanalization of CTO was successful. Our preliminary findings suggest that successful recanalization of CTO may have favorable outcome on left ventricular perfusion and function, particularly in patients with viable myocardium before PCI. The gated SPECT myocardial perfusion imaging with 99mTc-methoxy-isobutyl-isonitrile may be useful for monitoring long-term functional outcome of PCI in patients with CTO. PMID- 19306517 TI - Clinical investigations of 99mTc-p-aminohippuric acid as a new renal agent. AB - OBJECTIVE: 99mTc-p-aminohippuric acid (PAH) is a new radiopharmaceutical rapidly secreted by the kidneys in a manner consistent with tubular secretion. A comparative study of renal scintigraphy and clearance with 99mTc-PAH, diethylene triamine pentaacetic acid (DTPA) and mercaptoacetyltriglycine (MAG,) was performed. METHODS: 99mTc-PAH was prepared from a lyophilized kit by addition of sodium pertechnetate in the presence of DTPA. Ten healthy individuals were injected with 110 MBq of 99mTc-PAH. A dynamic study was repeated with 99mTc-DTPA and 99mTc-MAG3 several days later, after a 1-day interval. Clearance measurements were performed in five individuals. RESULTS: The mean values of time-to-peak activity (Tmax) and the time from peak to 50% of peak activity (T(1/2)) for 99mTc PAH (3.6 +/- 0.9 and 6.9 +/- 2.7 min, respectively) and 99mTc-MAG3 (3.5 +/- 0.8 and 6.8 +/- 2.1 min, respectively) were significantly lower in comparison with those of s99mTc-DTPA (4.9 1.7 and 11.7 +/- 1.9 min, respectively). The mean value of 99mTc-PAH clearance (186.9 +/- 12.2 ml/min) was significantly lower in comparison with MAG3 clearance (303.9 +/- 19.5 ml/min) and significantly higher than DTPA clearance (85.0 +/- 24.1 ml/min). CONCLUSION: Our preliminary results indicate the potential usefulness of 99mTc-PAH for routine renal scintigraphy. Owing to its fast kinetics, excretion properties and high-quality images, it could be a suitable substitute for 99mTc-DTPA. 99mTC-PAH clearance values, however, were significantly lower than those of MAG3, and could not be used for the estimation of renal plasma flow. PMID- 19306519 TI - When your belly hurts. PMID- 19306520 TI - New ban on plastic chemical. PMID- 19306521 TI - Controversies and advances in neonatal neurology: introduction. Introduction. PMID- 19306522 TI - [Announcement of retraction]. PMID- 19306523 TI - "For some queer reason": the trials and tribulations of Colonel Barker's masquerade in Interwar Britain. PMID- 19306525 TI - Childhood mortality in Central Spain, 1790-1960: changes in the course of demographic modernization. PMID- 19306524 TI - Clinical translation of genetic predictors for type 2 diabetes. AB - PURPOSE OF REVIEW: To highlight recent type 2 diabetes (T2D)-associated genetic discoveries and their potential for clinical application. RECENT FINDINGS: The advent of genome-wide association screening has uncovered many loci newly associated with T2D. This review describes the techniques applied to discover novel T2D genes and compares their relative strengths, biases, and findings to date. The results of large-scale genome-wide association studies carried out since 2007 are summarized, and limitations of interpreting this preliminary data are offered. Recent studies exploring the clinical potential of these discoveries are reviewed, focusing on insights into T2D pathogenesis, risk prediction of future diabetes, and utility in guiding pharmacotherapy. The new T2D-associated loci have been implicated in beta-cell development and function, highlighting insulin secretion in the disease process. Preliminary risk prediction studies show that more loci are needed to improve T2D risk indices. Studies have also revealed that genes may play a role in the pharmacologic response to antidiabetic medications. SUMMARY: Since 2007, genome-wide association studies have rapidly increased the number of T2D-associated loci. This review summarizes the history of genetic association studies, the results from the new genome-wide association studies, and the clinical application of these findings. PMID- 19306526 TI - Approach to the diagnosis and management of lipoprotein disorders. AB - PURPOSE OF REVIEW: Disorders of lipoprotein metabolism are frequently encountered in clinical practice. Although the severe genetic hyperlipidemias are relatively infrequent, prompt recognition and treatment can prevent complications, such as atherosclerosis and pancreatitis. The secondary dyslipidemias, due to medication or other metabolic disorders (hypothyroidism, renal or hepatic diseases), must be identified and treated. With the growing epidemic of obesity, dyslipidemias are a component of the metabolic syndrome. RECENT FINDINGS: The stratification of cardiovascular risk now includes family history and biomarkers of inflammation, especially high-sensitivity C-reactive protein, which enables sound clinical decision making. Lifelong hypercholesterolemia is strongly associated with increasing risk of atherosclerosis and coronary heart disease death, but the decision to treat pharmacologically depends on the absolute cardiovascular risk over the next 10 years. Clinical trial data support intensive treatment of patients at high cardiovascular risk or for the secondary prevention of recurrent coronary heart disease. The recently published JUPITER trial shows that patients with an elevated C-reactive protein benefit from treatment with a statin (rosuvastatin 20 mg) for primary prevention. SUMMARY: The current guidelines for the prevention of coronary artery disease will continue to focus on the determination of global risk, with intensive treatment aimed at the high-risk group. Family history and high-sensitivity C-reactive protein provide additional risk stratification. PMID- 19306527 TI - Dysfunctional high-density lipoprotein. AB - PURPOSE OF REVIEW: To address the progress of the investigation on dysfunctional high-density lipoprotein (HDL). RECENT FINDINGS: HDL is generally considered to be an independent protective factor against cardiovascular disease. However, emerging evidence indicates that HDL can be modified under certain circumstances and lose its protective effect or even become atherogenic. The underlying mechanisms responsible for generating the dysfunctional HDL and the chemical and structural changes of HDL remain largely unknown. Recent studies focus on the role of myeloperoxidase in generating oxidants as participants in rendering HDL dysfunctional in vivo. Myeloperoxidase modifies HDL in humans by oxidation of specific amino acid residues in apolipoprotein A-I, which impairs cholesterol efflux through ATP-binding cassette transporter A1 and contributes to atherogenesis. SUMMARY: HDL may not always be atheroprotective and can be atherogenic paradoxically under certain conditions. The mechanisms responsible for generating the dysfunctional HDL remain largely unknown. Recent data suggest that myeloperoxidase-associated modification of HDL may be one of the mechanisms. Further studies are needed to investigate the in-vivo mechanisms of HDL modification and identify therapeutic approaches aiming at controlling HDL modification. PMID- 19306528 TI - Lecithin: cholesterol acyltransferase--from biochemistry to role in cardiovascular disease. AB - PURPOSE OF REVIEW: We discuss the latest findings on the biochemistry of lecithin : cholesterol acyltransferase (LCAT), the effect of LCAT on atherosclerosis, clinical features of LCAT deficiency, and the impact of LCAT on cardiovascular disease from human studies. RECENT FINDINGS: Although there has been much recent progress in the biochemistry of LCAT and its effect on high-density lipoprotein metabolism, its role in the pathogenesis of atherosclerosis is still not fully understood. Studies from various animal models have revealed a complex interaction between LCAT and atherosclerosis that may be modified by diet and by other proteins that modify lipoproteins. Furthermore, the ability of LCAT to lower apoB appears to be the best way to predict its effect on atherosclerosis in animal models. Recent studies on patients with LCAT deficiency have shown a modest but significant increase in incidence of cardiovascular disease consistent with a beneficial effect of LCAT on atherosclerosis. The role of LCAT in the general population, however, has not revealed a consistent association with cardiovascular disease. SUMMARY: Recent research findings from animal and human studies have revealed a potential beneficial role of LCAT in reducing atherosclerosis but additional studies are necessary to better establish the linkage between LCAT and cardiovascular disease. PMID- 19306529 TI - The ABCG5 ABCG8 sterol transporter and phytosterols: implications for cardiometabolic disease. AB - PURPOSE OF REVIEW: This review summarizes recent developments in the activity, regulation, and physiology of the ABCG5 ABCG8 (G5G8) transporter and the use of its xenobiotic substrates, phytosterols, as cholesterol lowering agents in the treatment of cardiovascular disease. Recent progress has significant implications for the role of G5G8 and its substrates in complications associated with features of the metabolic syndrome. RECENT FINDINGS: Recent reports expand the clinical presentation of sitosterolemia to include platelet and adrenal dysfunction. The G5G8 sterol transporter is critical to hepatobiliary excretion of cholesterol under nonpathological conditions and has been linked to the cholesterol gallstone susceptibility. Finally, the cardiovascular benefits of cholesterol lowering through the use of phytosterol supplements were offset by vascular dysfunction, suggesting that alternative strategies to reduced cholesterol absorption offer greater benefit. SUMMARY: Insulin resistance elevates G5G8 and increases susceptibility to cholesterol gallstones. However, this transporter is critical for the exclusion of phytosterols from the absorptive pathways in the intestine. Challenging the limits of this protective mechanism through phytosterol supplementation diminishes the cardioprotective benefits of cholesterol lowering in mouse models of cardiovascular disease. PMID- 19306530 TI - Dead adipocytes and metabolic dysfunction: recent progress. AB - PURPOSE OF REVIEW: Dead adipocytes are at the heart of crown-like structures (CLSs), which represent a still relatively novel interface of adipocytes and adipose tissue macrophages (ATMs) and may be significant to human health conditions such as obesity, insulin resistance, diabetes mellitus, atherosclerosis, and cardiovascular disease. In order to provide a concise and pertinent discussion, this review highlights recent reports examining CLSs and projects the likely directions of future research in this budding and fascinating field. RECENT FINDINGS: In mice, there is recent progress in understanding the differences in distribution of two distinct phenotypes of ATM classes with respect to dead adipocytes. Furthermore, a detailed atlas of fat depot-specific differences in adipocyte size and CLS prevalence has begun in two genetic models of mouse obesity. In humans, fat cell aging and turnover have been quantified. The correlation between human CLSs and early cardiovascular disease has also been established. The predominant ATM phenotype in obese humans may be M2, which would contrast with the M1 ATMs of obese mouse models. Finally, osteopontin has been established in both humans and mice as an important macrophage chemoattractant to dead adipocytes, in addition to monocyte chemoattractant protein 1 and C-C motif chemokine receptor 2. SUMMARY: Recent updates in research on dead adipocytes and the biology of CLSs with clinical implications for metabolic dysfunction are discussed. PMID- 19306531 TI - [Physiological interactions: therapeutic tools in physiopathological constructions of the exophthalmic goiter, 1860-1960]. PMID- 19306532 TI - [Gymnastics and eugenicist ideology in France in the first half of the 20th century]. PMID- 19306533 TI - [Women, sports, identity, and acculturation during the first half of the 20th century]. PMID- 19306535 TI - Deep tissue injury: an evolving science. PMID- 19306534 TI - Bergstrand, Sandklef and the Frillesas mark. PMID- 19306536 TI - Wound tunneling. PMID- 19306537 TI - Calculating your patient's caloric needs. PMID- 19306538 TI - The cocoon strategy. PMID- 19306539 TI - The greatest generation? PMID- 19306540 TI - In defense of medicine. PMID- 19306541 TI - Identifying the identifier. PMID- 19306542 TI - Shots for big tots. PMID- 19306543 TI - Scaring up patients. PMID- 19306544 TI - Comparing the financial condition of Texas hospitals using a novel definition for the safety net. AB - Lack of health insurance is more prevalent in the state of Texas than in the rest of the country. To get necessary medical care, uninsured Texans must rely on safety net hospitals. Economic turmoil and fluctuating public support routinely threaten the financial stability of these hospitals. Safety net hospitals must be identified to craft public policy solutions that ensure their viability. In this paper, we propose a new method to identify these hospitals by incorporating criteria established previously by economists with additional measures of community value. Our data indicate that safety net hospitals continue to face financial challenges. Texas will need to move forward along several policy fronts to preserve this vital system of care. PMID- 19306545 TI - [The bible and pregnancy in other people]. PMID- 19306546 TI - ["Let nurses be visible in all their dimensions"]. PMID- 19306547 TI - [From now on a nurse talks in the name of all nurses]. PMID- 19306548 TI - [National organization of nurses, 23 regions, 23 presidents (4/6)]. PMID- 19306549 TI - [Patient information, in advance of good practices]. PMID- 19306550 TI - [Pain, work and nurses]. PMID- 19306552 TI - [Soon there will be a European chart for health professionals]. PMID- 19306551 TI - [Account of information on the law of Leonetti concerning the rights of patients at the end of life]. PMID- 19306553 TI - [Universal values concerning bioethics and cultural diversity]. PMID- 19306554 TI - [Surveillance of cross infections affects public health programs]. PMID- 19306555 TI - [National Federation of Nurses reforms its cooperation with physicians and nurses]. PMID- 19306556 TI - [Therapeutic education of type 2 diabetes patients, an approach whose object is learning about the digestive system]. PMID- 19306557 TI - [Better prevention and care of sores. Sores are an unfortunate preoccupation that is always present]. PMID- 19306558 TI - [Scientific publications about sores; what is new in 2008?]. PMID- 19306559 TI - [Supports to help the prevention and treatment of sores]. PMID- 19306560 TI - [Complications o sores]. PMID- 19306561 TI - [Taking care of sores by establishing by establishing facilities for dependent aged people]. PMID- 19306562 TI - [Scars at the end of life, a palliative approach]. PMID- 19306563 TI - [Palliative care of sores, from hospital to home]. PMID- 19306564 TI - [Negative pressure therapy, an essential complement to care of surgical scars]. PMID- 19306565 TI - [Care of scars, alternatives to hospitalization]. PMID- 19306566 TI - [1/10. Nursing profession. a century of becoming a profession]. PMID- 19306567 TI - [A patient with aphasia]. PMID- 19306568 TI - [3/5, The mission of nurses within the framework of the law of Leonetti]. PMID- 19306569 TI - [Heat shock proteins of the hyperthermophilic archaea]. AB - As thermostable enzymes and organisms are much more needed, researches on heat shock proteins(HSPs) of hyperthermophilic archaea have drawn more concerns. HSPs from hyperthermophilic archaea are concise only with HSP60, sHSP, prefoldin and AAA+proteins, but without HSP100s, HSP90s, HSP70 (DnaK), HSP40 (DnaJ) and GrpE which are common in mesophilic or thermophilic archaea. Accordingly, studies on the structure, function and operation mechanism of these four groups are much more important and meaningful. This review focuses on the recent progress in the researchs on the structure, function, operation mechanism and cooperation of the HSPs from hyperthermophilic archaea. The problems and obfuscations in these HSPs are analyzed, and farther research direction and key points are put out. PMID- 19306570 TI - [Sustainable production of bulk chemicals by application of "white biotechnology"]. AB - Practically all organic chemicals and plastics are nowadays produced from crude oil and natural gas. However, it is possible to produce a wide range of bulk chemicals from renewable resources by application of biotechnology. This paper focuses on White Biotechnology, which makes use of bacteria (or yeasts) or enzymes for the conversion of the fermentable sugar to the target product. It is shown that White Biotechnology offers substantial savings of non-renewable energy use and greenhouse gas emissions for nearly all of the products studied. Under favorable boundary conditions up to two thirds (67%) of the current non-renewable energy use for the production of the selected chemicals can be saved by 2050 if substantial technological progress is made and if the use of lignocellulosic feedstocks is successfully developed. The analysis for Europe (E.U. 25 countries) shows that land requirements related to White Biotechnology chemicals are not likely to become a critical issue in the next few decades, especially considering the large unused and underutilized resources in Eastern Europe. Substantial macroeconomic savings can be achieved under favourable boundary conditions. In principle, natural bacteria and enzymes can be used for White Biotechnology but, according to many experts in the fields, Genetically Modified Organisms (GMO) will be necessary in order to achieve the high yields, concentrations and productivities that are required to reach economic viability. Safe containment and inactivation of GMOs after release is very important because not all possible implications caused by the interaction of recombinant genes with other populations can be foreseen. If adequate precautionary measures are taken, the risks related to the use of genetically modified organisms in White Biotechnology are manageable. We conclude that the core requirements to be fulfilled in order to make clear steps towards a bio-based chemical industry are substantial technological progress in the bioprocess step and in downstream processing, high prices for fossil fuels and low prices for fermentable sugar. We strongly recommend to develop an integrated White Biotechnology strategy taking into account these four core requirements and other important accompanying activities. PMID- 19306571 TI - [Cloning and function analysis of the rice small GTP-binding protein gene OsPra2]. AB - Gene expression in rice roots under nutritional stress was studied using micro array techniques. The results showed that when re-supplied with sufficient amounts of nutrition after nutrition stress, the expression of OsPra2 (a small G protein which is homologous with Pea Pra2 protein) decreased in the plants root tissue. The cDNA sequence of the OsPra2 gene and its promoter, which is about 1 kb upstream of the translation origin point, was obtained using RT-PCR and PCR approaches. The OsPra2 protein contains four conserved GTP/GDP binding domains and specific domain of Rab small G protein family. The expression of OsPra2 and GST fusion protein in onion epidermal cells showed that OsPra2 protein was localized in the membrane and nucleus of the cell. The fusion expression of OsPra2 promoter and GUS reporter gene in transgenic rice suggested that the OsPra2 promoter allowed GUS expression in coleoptiles and roots. Compared with wild type rice, OsPra2 over expressed transgenic rice showed an obvious dwarf phenotype which resembles the BR deficient rice. PMID- 19306572 TI - [Construction and evaluation of a genetic engineered strain for biodesulfurization]. AB - We first cloned the dsz operon of Pseudomonas delafieldii R-8 into the expressing plasmid (pPR9TT) to construct the recombinant plasmid pPR-dsz, and then reintroduced it into strain R-8 to obtain a muti-copy dsz operon engineering strain R-8-1. Compared with the wild-type, strain R-8-1 showed a higher desulfurization activity for dibenzothiophene (DBT). Initial rates of DBT removal by strain R-8-1 were 6.25 micromol/g dry cell/h, about 2-fold higher than that for wild-type strain. The recombinant cells were also applied in the desulfurization of diesel. It resulted in a 68% reduction of total sulfur from 310.8 mg/L to 100.1 mg/L, whereas only 53% of sulfur was removed by strain R-8. The stability of pPR-dsz in strain R-8-1 was studied. The results revealed the first obtain a muti-copy dsz operon engineering strain are helpful for further development in biodesulfurization. PMID- 19306573 TI - [Cloning of silkworm 3-hydroxyisobutyrate dehydrogenase gene and its expression patterns analysis in simulated weightless environment]. AB - The full length cDNA of silkworm hibadh gene was cloned by RT-PCR and RACE (Rapid amplification of cDNA ends) technique. The hibadh gene and its deduced amino acid sequences were analyzed. The tissue distribution of hibadh gene in 5th instar silkworm larvae was tested by RT-PCR. The expression patterns of hibadh gene in simulated weightless environment were analyzed by real time RT-PCR. The results showed that the full length hibadh cDNA sequence was 1074 bp in lenth, including an open read frame of 969 bp encoding the entire coding region of Hibadh (GenBank accession No. EU719652). The deduced amino acid sequence similarities of hibadh between silkworm and Burkholderia ambifaria, Drosophila melanogaster, Apis mellifera, Xenopus tropicalis, Mus musculus, Homo sapiens were 46%, 43%, 48%, 44%, 45%, 45%, respectively. Signal peptide analysis showed that Hibadh was a secretory protein. There wasn't glycosyl-phosphatidyl inositol anchor site in Hibadh amino acid sequence. Molecular weight and isoelectric point of Hibadh were 34.1 kD and 9.14 respectively. The RT-PCR tests indicated that the hibadh gene expressed in head, silk gland, midgut, cuticle, blood, fat body, tuba malpighii of the 5th instar silkworm larvae. There were different expression patterns of hibadh gene during different silkworm embryo period in simulated weightless environment. Simulated weightlessness resulted in the expression of silkworm hibadh gene up regulated 2.3-fold (P < 0.05), up regulated 4.6-fold (P<0.01), down regulated 7.6-fold (P < 0.01), down regulated 2.6-fold (P < 0.05) during apophysis formation period, inverse period, trachea formation period, and whole embryo period, respectively. There was no significant change of hibadh gene expression during other period of silkworm embryo between simulated weightless and control groups. There were different response patterns to simulated weightless environment between hibadh gene and whole body of silkworm. Gene showed much higher sensitivity compared to whole body in response to environment. This study is useful for the further research on the gravity biological mechanism of hibadh gene. PMID- 19306574 TI - [Preparation and identification of recombinant maxadilan]. AB - To produce recombinant Maxadilan using gene engineering technology, the gene of recombinant Maxadilan which expressed in protocaryon were designed and synthesized according to the amino acid sequences of Maxadilan. The recombinant plasmid pKYB-MAX was constructed and transformed into host bacteria Escherichia coli strain ER2566. After the MAX-intein-CBD fusion protein was purified by chintin-affinity chromatography, the self-cleavage activity of the intein was induced by beta-mercaptoethanol and the recombinant Maxadilan was released from the chitin-bound intein tag. The molecular weight of peptides was determined by the laser flight mass spectrometry and the results was conformity with the theoretical value. The biological activity analysis showed that recombinant Maxadilan significantly enhanced the concentration of serum glucose. PMID- 19306575 TI - [Establishment of fibroblast cell line and its biological characteristics in Matou goat]. AB - Taking Matou goat ear margin as the study material, we succeeded in established a fibroblast cell line by the method of explant culture directly. Observations on morphology, dynamic growth, determination of viability, analysis of karyotype, test of microorganism and other characteristics were detected. Results showed: Population Doubling Time (PDT) of cells was approximately 36 h; Cell viability was 96.7% after thawing; The status of cell After passage was constant; Analysis of chromosomal karyotyps indicated that diploid (2n=60) account for 98% in the cell line. Every index in the cell line met all the standard quality controls of ATCC in USA. The established of Matou goat ear fibroblast cell line has not only important genetic resources preserved at the cell level, but also valuable material for genome, postgenome and somatic cell nuclear transfer research. PMID- 19306576 TI - [Construction of lentiviral vectors encoding GDNF and GDNF expression in human neural stem cells]. AB - We transfected human neural stem cells using lentiviral vectors encoding glial cell line derived neurotrophic factor (GDNF) to study its expression level in vitro and to get a stable cell line expressing GDNF. First, GDNF gene was sub cloned into the lentiviral transfer vectors. Then, the recombinant lentiviral supernatants were packaged by 293T cells through three plasmids transient co transfection method using standard lipofectamine reagent. The viral titers were tested by the transfection efficiency of 293T cells. At the same time, human neural stem cells (hNSC) were transfected under different multiplicity of infection. GDNF gene expression level and protein secretion level of hNSC were tested by real-time PCR and ELISA methods after transfection. Lentiviral vectors encoding GDNF were constructed. Using lentiviral vectors encoding GDNF we successfully transfected human neural stem cells, and got a stable neural stem cell lines over-expressing GDNF. Furthermore, the results indicated that GDNF expression was influenced by the multiplicity of infection. Human neural stem cells could over-express GDNF through lentivial vectors tranfection. Its gene expression level and protein expression level correlate with the multiplicity of infection. PMID- 19306577 TI - Purification and characterization of extracellular laccase secreted by Pleurotus sajor-caju MTCC 141. AB - The effect of lignin containing natural substrates corn-cob, coir-dust, saw-dust, wheat straw and bagasse particles on the extracellular secretion of laccase in the liquid culture growth medium of Pleurotus sajor-caju MTCC 141 has been studied. The culture conditions for maximum secretion of laccase by Pleurotus sajor-caju MTCC 141 have been optimized. Homogeneous preparation of laccase from the culture filtrate of the fungus has been achieved using ammonium sulphate precipitation, anion exchange chromatography on DEAE and gel filtration chromatography on Sephadex G-100. The purified enzyme preparation gave a single protein band in SDS-PAGE analysis indicating a molecular weight of 90 kD. The enzymatic characteristics Km, k(cat), pH and temperature optima of the purified laccase have been determined using 2, 6-dimethoxyphenol as the substrate and have been found to be 35 micromol/L, 0.30 min(-1), 4.5 and 37 degrees C respectively. The Km values for the other substrate like catechol, m-cresol, pyrogallol and syringaldazine have also been determined which were found to be 216 micromol/L, 380 micromol/L, 370 micromol/L and 260 micromol/L respectively. PMID- 19306578 TI - Purification and properties of cold-active metalloprotease from Curtobacterium luteum and effect of culture conditions on production. AB - Curtobacterium luteum, a gram-positive psychrotrophic bacterium, secreting an extracellular protease was isolated from the soil of Gangotri glacier, Western Himalaya. The maximum enzyme production was achieved when isolate was grown in a pH-neutral medium containing skim milk at 15 degrees C over 120 hour. The metal ions such as Zn2+ and Cr2+ enhanced enzyme production. The specific activity of purified enzyme was 8090 u/mg after 34.1 fold purification. The 115 kD enzyme was a metalloprotease (activity inhibited by EDTA and EGTA) and showed maximum activity at 20 degrres C and pH 7. The enzyme was active over a broad pH range and retained 84% of its original activity between pH 6-8. There was no loss in enzyme activity when exposed for 3 hours at 4 degrees C-20 degrees C. However, lost 65% of activity at 30 degrees C, and was almost inactivated at 50 dgrees C, but was resistant to repeated freezing and thawing. The enzyme activity was stimulated by manganese ions; however, it was inactivated by copper ions. PMID- 19306579 TI - [Construction of engineered Escherichia coli for aerobic succinate production]. AB - Based on carbon metabolic pathway analysis of Escherichia coli MG1655, an aerobic succinate fermentation platform was constructed by knocking out five genes (ptsG, poxB, pta, iclR and sdhA), which was named E. coli QZ1111. Flask cultivation results showed that E. coli QZ1111 could accumulate succinate with a concentration of 26.4 g/L under aerobic conditions. The byproduct acetate was only 2.3 g/L. The production ratio of succinate and acetate reached 11.5:1. PMID- 19306580 TI - [Acylation specificity of midecamycin 3-O-acyltransferase within Streptomyces spiramyceticus F21]. AB - Spiramycin and midecamycin are 16-membered macrolide antibiotics with very similar chemical structures. Spiramycin has three components, namely spiramycin I, II and III. Spiramycin II and III are, respectively, the O-acetyl and propionyl derivatives at C3-hydroxyl group of spiramycin I. Midecamycin has four components, and the C3-hydroxyl group of midecamycin is all O-propionylated. The enzyme adding acyl group(s) at the C3-hydroxyl group during the biosynthesis of spiramycin and midecamycin is 3-O-acyltransferase. The 3-O-acyltransferases for spiramycin and midecamycin are also very similar, and presume to function when exchanged. To explore whether the 3-O-acyltransferase for midecamycin biosynthesis hold still the character of selective and efficient propionylation for spiramycin I at its C3-hydroxyl group, we inserted mdmB, the 3-O acyltransferase gene from Streptomyces mycarofaciens ATCC 21454 for midecamycin biosynthesis, into a mutant strain of S. spiramyceticus F21, in which the 3-O acyltransferase gene for spiramycin biosynthesis, sspA, was deleted; and the mdmB was integrated exactly into the chromosomal site where the sspA was deleted. We name this "hybrid" strain as SP-mdmB. HPLC analysis of the spiramycin produced by SP-mdmB showed that spiramycin I was still the major component, although the relative proportions of both spiramycin II and III increased significantly. We thus conclude that MdmB from Streptomyces mycarofaciens ATCC 21454 for midecamyicn biosynthesis do not hold the character of selective and efficient propionylation for spiramycin I within S. spiramyceticus F21, and this character is possibly limited in Streptomyces mycarofaciens ATCC 21454 for midecamycin biosynthesis. PMID- 19306581 TI - [Bioassay of recombinant green fluorescent protein gene yeast cell for a high throughput to screen estrogenic compounds]. AB - We developed the recombinant green fluorescent protein gene yeast cell to screen estrogenic compounds based on two episomal vectors. In the expression vector the expression of human estrogen receptor alpha(hERalpha) was driven by 3 glyceraldehydephosphate dehydrogenase (GPD) promoter; in the reporter vector the expression of the yeast enhanced green fluorescent protein (yEGFP) gene was under the control of the estrogen response element (ERE). The vectors were transformed into yeast cell (W303-1A) to construct GFP recombinant yeast cell. Incubation of the yeast cell with various concentrations of the estrogenic compounds led to expression of the reporter gene product GFP in a dose dependent manner. Compared to other yeast bioassays, the yeast cell for environmental estrogen bioassay based on yEGFP reporter gene did not need cell wall disruption or the addition of a substrate or reagent. This yEGFP assay was performed completely in 96 well plates. So this test system can be used as a rapid and high throughput system for screening estrogenic chemical products, which has the characteristics of the sensitivity, reproducibility and cheapness. PMID- 19306582 TI - Optimization of fermentation conditions for maximal recombinant hepatitis B surface antigen particle production in Pichia pastoris. AB - Hepatitis B virus (HBV) infection can cause the severe threat to the health of the people around the world. It depends upon the development of efficient diagnostic reagent and vaccine to prevent the prevalence of HB. In this study, we constructed the high expression recombinant Pichia pastoris and performed the screening tests in shake flasks to obtain the optimal values of several key fermentation parameters. Based on their effects on the growth and expression level of recombinant strains, FBS was the optimal industrial medium. The optimal values for the dissolve oxygen (DO), the final concentrations of methanol and the pH values were 50 mL, 1% (V/V) and 5.4-6.0, respectively. The optimal values of the parameters simulated in shake flasks were successfully scaled up to 10 L bioreactors to achieve high-throughput production: 310 OD600 in biomass and 27 mg/L in recombinant HBsAg. The expressed recombinant HBsAg in P. Pastoris was confirmed by SDS-PAGE and Western blotting. Electron microscopy examination showed that the purified protein could be self-assembled to 22 nm virus-like particles. The results provided a basis for industrial scale-up production of diagnostic reagent and vaccine of next generation against HB. PMID- 19306583 TI - [Functional analysis of specific promoter using vecotors harboring GFP/RFP double fluorescent marker genes]. AB - Most studies related to determining the expression profile of genes and specific promoters used histochemical localization of the reporter gene, gusA. While the histochemical method for visualizing gusA expression suffers from several limitations in the determination of gene expression and location, especially in the tissues with high background acitivty. To solve this problem, a transient expession vector pBI221-RFP/GFP, was constructed using GFP and RFP as double fluorescent marker genes. This vector used CaMV 35S promoter to drive GFP and determine the transforming efficiency. It analyzed expression profile of the target gene and promoter through the RFP activities of the tranformed tissues. Through the specific promoter AGPL1 from watermelon and E8 promoter from tomato, it is resistible to use this vector to study the expression patterns of promoters. Results indicated that the pBI221-RFP/GFP is a very efficient transient expression vector that can be verify the functions of the genes and promoters. PMID- 19306584 TI - [Numerical simulation of dental composite damage caused by water sorption]. AB - Damage effects of water sorption on mechanical properties of the hydroxyapatite particle reinforced Bis-GMA/TEGDMA copolymer (HA/Bis-GMA/TEGDMA) have been predicted using 3-D finite cell models. Three different cell models were used to determine the influence of varying particle contents, interphase strength and moisture concentration on the debonding damage. The stress distribution pattern has been examined and the stress transfer mode has been clarified. The Young's modulus and fracture strength of the Bis-GMA/TEGDMA composite were also predicted using the model with and without consideration of the damage. The former results with consideration of the debonding damage are in good agreement with existing literature experimental data. The shielding effect of our proposed model and an alternative approach were discussed. The FCC cell model has also been extended to predict the critical load for the damaged and the undamaged composite subject to the 3-point flexural test. PMID- 19306585 TI - [Purification of large-scale plasmid DNAs by selective precipitation with cetyltrimethylammonium bromide]. AB - Following Escherichia coli lysis with alkali, cetyltrimethylammonium bromide (CTAB) was directly titrated into the supernatant. An easy and feasible technology for plasmid purification was established with the optimized proportion between the quantity of CTAB and plasmid, combined with the specific solution for DNA release and TritonX-114 for endotoxin removal. Quality detection showed that the purified plasmid was free of contamination of host RNA. The host bacterial genomic DNA, endotoxin and bacterial protein were less than 10 microg, 50 EU and 10 microg per mg plasmids, respectively. The ratio of OD260/OD280 was between 1.75-1.85. Eighty percent of the prepared plasmids were presented in the supercoiled form. The plasmid purified with this technology can satisfy all criteria stipulated by FDA. The main advantages of the technology include the avoidance of animal-derived enzymes such as ribonucleases A, Proteinase K and toxic reagents like chloroform and phenol. In addition, the technology has low cost and no pollution. PMID- 19306586 TI - Culture and race in provider-client relationships. AB - Given that minority group members are underrepresented in the teaching, medical, and legal professions, minority group members often have White teachers, doctors, and lawyers. This is frequently the case even when students, patients, and clients would prefer service providers similar to them in racial or ethnic background. This paper identifies possible cultural barriers to effective one-on one relationships between White teachers, doctors, and lawyers and those who receive their services; explores the potential for biased expectations to influence the services provided and outcomes attained; and contrasts the goals of White and minority educators, doctors, and lawyers, arguing that these differences have potentially negative implications for minority service recipients. Policy approaches to mitigating potential problems caused by the lack of match are considered, as are potential problems arising from an overly narrow emphasis on match. PMID- 19306587 TI - Reconceptualizing access: a cultural competence approach to improving the mental health of African American women. AB - Despite the prevalence of mental illness among African American women, only a limited number of them seek or accept help from mental health service delivery systems. An extensive review of the literature revealed that (1) racism and discrimination, (2) socioeconomic status, (3) stress and well being, and (4) housing and neighborhood conditions must be considered in an assessment of the mental health status of African American women. These factors negatively affect their mental health and should be addressed in eliminating disparities in access to and utilization of mental health services. We recommend a process by which mental health providers reconceptualize access to mental health services using a socio-cultural framework. The knowledge gained in this process will result in increased provider cultural competence. This developmental process would be facilitated by the use of a socio-cultural conceptual model for treatment engagement. The model takes into consideration the barriers to mental health treatment services that, in part, have to be eliminated by mental health providers in order to decrease disparities and enhance both access to and utilization of mental health services by African American women. PMID- 19306588 TI - Depression in African American and White women with low incomes: the role of chronic stress. AB - The current study examined the links between frequency and severity of acute and chronic stress, on the one hand, and depressive symptoms, on the other hand, in a racially balanced sample of African American and White women with low incomes. We predicted and found that severity of chronic stress accounted for more of the variance in depressive symptoms than did severity of acute stress and that severity of chronic stress amplified the effects of the severity of acute stress on depressive symptoms. Results also showed that African American women tended to have a greater number of depressive symptoms than White women, controlling for income, possibly because they experienced a greater number of chronic stressors than did White women, a condition that rendered them more vulnerable to psychological distress. PMID- 19306589 TI - Racial differences in mental health service utilization among low-income women. AB - The authors present data on mental health outpatient services in non-specialty settings in a sample of low-income women by exploring service use in the general medical sector, specialty mental health/substance services, the human service sector, and self-help groups. Findings are reported from 668 African American and White women in the Mothers' Well-Being Study (MWS). The MWS measured a range of psychiatric and substance dependence disorders using the Composite International Diagnostic Interview, Version 2.1 (CIDI2.1). The MWS also gathered data regarding outpatient mental health service utilization. In the year preceding the study, 43.9% of the White respondents and 39.0% of the African American respondents had at least one of the mental health disorders measured in the MWS. There were no significant differences in the frequency of any of the disorders by race. However, White respondents with disorders received more treatment than African American women in the general medical sector. In the year prior to the interview, nearly a quarter (22.4%) of White women with any diagnosis received care in the general medical sector compared to only 9.1% of African American women. The racial disparity in mental health treatment in the general medical sector may indicate that African American low-income women are not receiving the same level of care as White women. The authors discuss the implications of this disparity and suggest ways of expanding access to care for African American women in the general medical setting. PMID- 19306590 TI - African American consumers' perceptions of racial disparities in mental health services. AB - In 2001, former Surgeon General, Dr. David Satcher, reported that minorities in the United States suffer a disproportionate burden of mental illness because of a large gap between the need for mental health services and the services actually provided. While research findings and policy analyses show that racial disparities exist, there has been little exploration of how minority recipients of mental health services perceive such disparities. This paper reports findings from a study using consumer focus groups with African Americans to explore how African American mental health consumers perceive and personally experience the impact of racial disparities in accessing and utilizing mental health services. Findings showed that the consumers voiced similar concerns to those reported by the Surgeon General; however, they also identified problems in communicating with providers as a major obstacle to seeking services and engaging in treatment. The consumers suggested a number of specific recommendations to improve services in their community. PMID- 19306591 TI - The need to adapt standardized outcomes measures for community mental health. AB - In order to determine whether evidence-based practices are effective in community settings, the use of measurement tools that accurately capture symptoms and functioning in the community context is imperative. Without accurate measurement, researchers face the troubling possibility that tests of effective treatments may not attain positive results because of measurement error. The authors have experienced apparently serious problems with measurement validity in community mental health practice settings in which we are conducting diffusion research. Features of survey design that strengthen measurement in the university setting (where survey instruments are designed and treatments are usually tested) appear to work against us in community settings. Problems are compounded when it is necessary to measure across several domains, and thus, to employ multiple questionnaires. Here, we discuss the problems we have experienced with three common survey design strategies: (1) the use of multiple, similar items; (2) the use of reverse-coded items; and (3) the use of graded response-items. These strategies result in measures whose length and cognitive complexity may compromise their validity in community settings. Race is an especially important issue in diffusion research, as we lack even basic information concerning efficacy and effectiveness for some important evidence-based treatments. Invalid measures could seriously delay transfer and development of appropriate treatments. Careful psychometric work is badly needed, but such work is time intensive and costly. In the meantime, we encourage researchers to consider the modifications suggested here. PMID- 19306592 TI - Lessons learned: the unintended consequences of policy decisions affecting maternity services for Pittsburgh's African Americans. AB - In the Pittsburgh region, as elsewhere, the Black infant mortality rate, reflecting overall health disparities, is higher than the rate for White babies. This study suggests that the answer to the infant mortality racial disparity question may lie partially in the characteristics and availability of prenatal care as well as broader changes in the social and economic environment. Numerous known risk factors associated with infant mortality have been present for as long as statistics have been recorded. Only wide ranging comprehensive programs will combat such factors as low income, education, teenage motherhood, unemployment, and inadequate housing. The other more specific factors of low birth weight and inadequate prenatal care, however, can be addressed by providing adequate prenatal care in a setting that is accessible and designed specifically with the cultural awareness and concerns of the African American community in mind. The strategies of fifty years ago offer some insights that are worth noting as policy planners begin to design and implement new programs to combat the continuing tragedy of African American infant mortality. PMID- 19306593 TI - Disparities in health status and health-service utilization among Hispanic ethnic subgroups. AB - Using nationally representative National Health Interview Survey (NHIS) data from 1999 through 2002, this study examines the health services access and utilization patterns of U.S. Hispanic adolescents under age 18 years classified as: Cubans, Puerto Ricans and Dominicans, Mexicans or Mexican Americans, Central and South Americans, and mixed Hispanic and non-Hispanic White. Consistent with previous studies on ethnic patterns in child and adult health, Hispanic American children do better than non-Hispanic Whites with respect to certain health status measures such as school absence. There is considerable heterogeneity in health status and health care utilization in the Hispanic subgroup, indicating the necessity of separating Hispanic subgroups for analytic purposes. PMID- 19306594 TI - Race/ethnicity, religiosity and adolescent alcohol, cigarette and marijuana use. AB - Previous research suggests that religiosity "protects" young people from substances abuse. Because most of this research has been based on samples of White youth, however, the extent to which its findings apply to Black and Hispanic youth is largely unknown. Therefore, the purpose of the present study is to address two relatively simple but essential questions: first, "How religious are American Black and Hispanic adolescents?" and second, "Does religiosity 'protect' Black and Hispanic young people from drug use as research suggests that it protects White youth?" The present study analyzed data from large, nationally representative samples of Black, Hispanic, and White 10th graders drawn from the Monitoring the Future Project. The results indicate that the majority of 10th graders, irrespective of racial or ethnic group, are at least somewhat religious and that a third or more might be considered very religious. The data further suggest that Black 10th graders are significantly more religious than White and Hispanic 10th graders. The data suggest that religion does, in fact, "protect" Black and Hispanic youth from substance abuse, but that the strength of this relationship is greater for White than for non-White youth. The reasons for racial and ethnic differences in the strength of the relationship between religiosity and substance abuse are not clear. One possibility is that religiosity may be more of a cultural or group phenomenon among non-White youth, while among White youth it may be more of an individual factor affecting individual behavior such as substance use. Understanding the mechanisms by which religion might influence substance use and the reasons why these mechanisms may vary by race and ethnicity is an important task for future research. PMID- 19306595 TI - Ways with children: examining the role of cultural continuity in early educators' practices and beliefs about working with low-income children of color. AB - New directions in child care research are compelling researchers to explore larger socialization perspectives. In one of these, ethnic and cultural matches between caregiver and child are used to explain important psychological and developmental processes in children of color (Guerra & Jagers, 1998; McLoyd, 1998; Phinney & Landin, 1998). This paper discusses three recent child care studies that have used teacher-child ethnicity as a proxy for cultural continuity. The first study identified teacher and program practices in child care programs designed to serve low-income children and families of color and used a teacher-child ethnic match to ground its examination of associations among teacher-articulated practices, quality, and observed child behaviors. The second study also used a teacher-child ethnic match to ground its examination of associations among teacher-articulated beliefs and practices about families and observed teaching practices in child care programs that served low-income children and families of color. The third study explored young children's processes of forming attachments with caregivers in child care when the child and caregiver did and did not share an ethnic cultural community. The findings in all three studies suggested that teachers' perceptions and practices, and children's experiences in child care, are rooted within ethnic communities. PMID- 19306596 TI - Minority business bidding for local government contracts: the complexity of availability. AB - While minority-business enterprises (MBEs) have gained some access to local government contracts during the last three decades, these firms continue to receive a small share of local government contract spending relative to the number of available firms. Researchers have suggested two general explanations for the low representation of MBEs in contract awards: (1) lack of qualifications and capacity among MBEs, and (2) public and private discrimination against MBEs in contracting processes. This study on prime contract opportunities in a Northern central city and county with a large minority population finds that low bid rates greatly contribute to the low MBE shares of prime contracts and that bidding is reduced by both local government processes and characteristics of the firms. Some implications of these findings are that local governments need to: (1) monitor MBE shares of prime contract bids by size of contract and use share of bids as one measure of program and organizational effectiveness; (2) identify MBEs that are qualified for prime contracts and encourage and help interested firms to submit competitive bids; and (3) ensure that local government policies and practices do not diminish access to information about prime contract opportunities for qualified and interested minority firms. Another implication is that bidders lists should not be a primary basis for determining MBE availability, since many qualified and interested MBEs do not bid because of perceived barriers in local government. PMID- 19306597 TI - [Casein phosphopeptide--amorphous calcium phosphate (CPP-ACP) and its effect on dental hard tissues]. AB - Dental products with casein phosphopeptide--amorphous calcium phosphate nanocomplexes (CPP-ACP) are used in several tooth products (toothpastes, chewing gums, mouthrinses) and are as well used in dental filling material. CPP-ACP containing products are supposed to enhance remineralisation of dental hard tissues und thus might play a major role in prevention and therapy of initial caries or erosively dissolved enamel. Furthermore, also in hypersensitive teeth and even cases of hyposalivation, CPP-ACP containig products are supposed to improve the clinical condition. This article aims at three goals: point out the evolvement of CPP-ACP out of milk casein; description of possible biochemical effects of CPP-ACP on dental hard tissues; critical review of the current literature. PMID- 19306598 TI - [Gingival retraction methods. A literature review]. AB - The exposure of the preparation margin and the control of the hemorrhage in the gingival sulcus are prerequisites for precise impressions and thereby improving the quality of indirectly fabricated restorations. The purpose of this review article is to summarize available evidence with respect to current methods of gingival retraction and to provide the clinician with practical tips. PMID- 19306599 TI - [Dental materials. A critical assessment from the viewpoint of alternative medicine]. AB - Practical experience with unconventional treatments suggests that the paradigm used by conventional medical science to assess the impact of dental materials must be broadened. First, several diagnostic methods commonly employed to evaluate allergies and toxicological burdens are described and subjected to a critical analysis. These diagnostics include test methods used in the field of complementary medicine in addition to the traditional epicutaneous tests, the Lymphocyte Transformation Test and quantitative analysis of blood and urine. Finally, the fundamentals of toxicology in the low-dose range are discussed; in this context special attention is paid to possible factors enhancing the effect of various substance groups. The impact of dental materials is also viewed from the perspective of environmental toxicology. In addition, the authors discuss various paradigms for obtaining evidence of multifactorial causes and show why nonuniform results are obtained with dental materials. Reference is also made to new theories broadening our understanding of biological processes such as the Biphoton Theory, which has been the subject of increased discussion among quantum physicists in recent years. It becomes evident in this context that there are to date no evidence-based methods for demonstrating the absolute non-toxicity of dental materials. Finally, it is shown - on the basis of various reports provided by a practitioner of complementary medicine in private practice - that, in patients with chronic diseases, unconventional therapies integrating these insights may by the only effective therapeutic options to succeed. PMID- 19306600 TI - [The role of the dentist in modern forensic age determination]. PMID- 19306601 TI - Cytomegalovirus vaccines: at last, a major step forward. AB - Yow and Demmler published an editorial in 1992 entitled Congenital cytomegalovirus disease--20 years is long enough. This editorial pointed out that, despite the long-standing recognition of the major public health importance of congenital human cytomegalovirus (CMV) infection, few solutions had been forthcoming. In no area has the lack of progress been more frustrating than in the realm of CMV vaccine development. Although a number of vaccine strategies have been considered, few have advanced to the stage of efficacy testing. PMID- 19306602 TI - Seroprevalence of herpes simplex virus-2 infection among women seeking medical care for signs and symptoms of vaginitis. AB - Two identically designed studies were conducted to determine the prevalence of herpes simplex virus type 2 (HSV-2) infection and viral shedding among women with no known history of genital herpes or HSV-2 seropositivity, who sought care at a US-based obstetrics/gynaecology clinic because of recurrent signs and symptoms of vaginitis. Samples comprised 50 women of any race (All-Comers Sample; Study 1) and 49 black women (Black Sample; Study 2) diagnosed at the clinic visit with vaginitis on the basis of standard work-up and medical history. In the All-Comers Sample, 15 (30%) women were HSV-2 seropositive and two (4%) were positive for HSV 2 by polymerase chain reaction (PCR); these two patients were also HSV-2 seropositive. Therefore, among patients seropositive for HSV-2, two (13%) were shedding virus at the time of the clinic visit. In the Black Sample, 25 (51%) were HSV-2 seropositive and two (4%) were PCR positive for HSV-2. Factors associated with HSV-2 seropositivity included age >30 years, erythema on pelvic examination, age < or = 15 years at first intercourse and having more than four lifetime sexual partners. The high prevalence of HSV-2 infection in these samples underscores the need to test for HSV-2 infection in women consulting with signs and symptoms of vaginitis. PMID- 19306603 TI - Genital herpes education and counselling: testing a one-page 'FAQ' intervention. AB - Approximately 17% of adults in the USA have antibodies to herpes simplex virus type 2 (HSV-2), 85-90% of whom are unaware of their infection. Diagnosed patients are more likely to detect HSV reactivation, disclose infection status to partner(s) and employ risk-reduction strategies; therefore, diagnosing more cases may reduce herpes transmission rates. Providers are reluctant to identify genital herpes because of potentially lengthy counselling visits or discomfort in dealing with patients' emotional reactions. This study tested the efficacy of a one-page frequently-asked-questions (FAQ) sheet regarding length of initial diagnostic visit, patient satisfaction with topics covered, provider's coverage of topics and resources/referrals provided to patients. Data from 147 patients and 26 providers were analysed. When pre- and post-FAQ sheet data were compared, diagnostic visit time was not substantially reduced but there were favourable trends in patient satisfaction levels on some topics, and providers covered more topics and offered more resources. This FAQ was useful in simplifying topics, providing prompts for providers and resources for patients. While this FAQ sheet appears favourable in this exploratory study, more investigation is needed to measure the degree to which patients use and understand the FAQ sheet, and find it to be a helpful tool. PMID- 19306604 TI - Common questions about herpes: analysis of chat-room transcripts. AB - Patients diagnosed with genital herpes typically undergo a period of psychological adjustment. Although healthcare providers can play a key role in this adjustment, in several patient surveys patients have expressed dissatisfaction with the information and counselling offered by professionals. To address this gap, providers must first identify the common questions and myths that are not addressed, or are addressed inadequately. This article is that first step. Through a content analysis of herpes chat-room transcripts captured on their website from autumn 2001 to spring 2006, researchers from the American Social Health Association identified common herpes questions and myths. The 1968 chat passages were coded into 12 themes and 50 sub-themes. Frequently, visitors' questions concerned transmission, symptoms and diagnosis followed by natural history, psychosocial issues and treatment options. The results of this analysis will aid in the creation of tailored messages to address common factual questions and provide psychosocial support. PMID- 19306605 TI - Varicella zoster virus redux. AB - In May 2008, the Advisory Committee on Immunization Practices (ACIP) of the Centers for Disease Control and Prevention in the USA recommended the routine administration of a live-attenuated high-potency varicella zoster vaccine to all adults over 60 years of age without specific contraindications, for the prevention and attenuation of herpes zoster (HZ). Nevertheless, many physicians still consider this vaccine to be of marginal value. This is not a reasonable conclusion. This short article reviews available data. PMID- 19306606 TI - Treatment for Epstein-Barr virus-associated PTLD. AB - The association between Epstein-Barr virus (EBV) and post-transplant lymphoproliferative disease (PTLD) has been recognized since the early days of transplantation. The major pathogenetic defect is the insufficient EBV-specific cytotoxic T-cell control of EBV-driven B-cell proliferations. Despite this understanding, PTLD remains a significant cause of morbidity and mortality for transplant recipients. Determining the right therapy or therapies for any given patient with PTLD remains a major clinical problem. Productive areas of investigation include: identifying who will benefit from reduction of immunosuppression only; improving methods to predict those at highest risk of PTLD; developing safe and effective pre-emptive therapies; identifying who will benefit from rituximab; and developing more effective, less toxic therapies for resistant or aggressive disease. Obstacles that exist are heterogeneity of disease and patient populations, and divergent approaches to immunosuppression and therapeutic interventions. Greater collaboration is needed between infectious disease specialists, pathologists, transplant physicians and oncologists to overcome problems and develop agreed disease definitions and interventions that can be tested in large, prospective multicentre trials. PMID- 19306607 TI - Improved method for determining antibiotic susceptibility of Flavobacterium columnare isolates by broth microdilution. AB - A simple and reproducible microdilution method was developed to test the susceptibility of the bacterium Flavobacterium columnare to antibiotics in vitro. The testing was conducted at 28 degrees C for 44-48 h at two dilutions of Mueller Hinton broth (DMHB) using a standardized inoculum, a reference isolate of Escherichia coli ATCC25922 as a quality control organism, positive and negative control wells, and standardized custom-made microtiter plates. The E. coli ATCC25922 and F. columnare ATCC23463 (the species type strain) had significantly better growth in DMHB at 1:5 (4 g/L) than at 1:7 (3 g/L). The E. coli ATCC25922 was found to be acceptable as a reference isolate and produced minimum inhibitory concentration (MIC) values similar to those in the range published by the Clinical and Laboratory Standards Institute derived using standard Mueller-Hinton broth. The new method was used to determine the MIC of 23 F. columnare isolates (representing the three genotypes of the species) to enrofloxacin, ampicillin, oxytetracycline, erythromycin, florfenicol, flumequine, ormetoprim/sulfadimethoxine, and oxolinic acid. PMID- 19306608 TI - Nested polymerase chain reaction assay for detection of Mycobacterium shottsii and M. pseudoshottsii in striped bass. AB - Wild striped bass Morone saxatilis in Chesapeake Bay are experiencing a high prevalence of mycobacteriosis, which produces granulomatous lesions of the skin and visceral organs. Culture-based studies have indicated that the newly described species Mycobacterium shottsii and M. pseudoshottsii are the dominant isolates from diseased fish. The classical fish pathogen M. marinum is also found, albeit at much lower frequencies. Both M. shottsii and M. pseudoshottsii are extremely slow-growing on standard selective media, and up to 12 months may be required for isolation and characterization. Epidemiological studies of mycobacteriosis in Chesapeake Bay would therefore benefit from rapid molecular assays with which to detect these species in fish. In this paper, we describe the development of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays capable of detecting M. shottsii, M. pseudoshottsii, and, in most instances, coinfections thereof in striped bass tissues. In addition, PCR RFLP assays were designed to detect M. marinum and other as-yet-undescribed Mycobacterium spp. present in Chesapeake Bay striped bass. Comparison of these molecular assays with culture-based techniques using splenic tissue from wild striped bass yielded generally concordant results and demonstrated the applicability of these techniques to the study of wild fish. PMID- 19306609 TI - Comparison of tank treatments with copper sulfate and potassium permanganate for sunshine bass with ichthyobodosis. AB - The biflagellated, single-celled parasite Ichthyobodo necator can cause significant losses among fish populations, particularly those cultured in tanks. Treatments of KMnO4 and CuSO4 were evaluated against a naturally occurring I. necator infestation on sunshine bass (female white bass Morone chrysops x male striped bass M. saxatilis) raised in tanks. Four-hour static treatments with 3 mg of KMnO4/L of water (2.5 mg/L above the determined KMnO4 demand) or 2 mg of CuSO4/L of water (total alkalinity = 207 mg/L; total hardness = 95 mg/L) were randomly applied to 4 tanks/treatment (23 fish/tank); the same treatments were reapplied 2 d later. Four tanks were used as positive controls. By 2 d posttreatment (after the second treatment), only 17.4% of the untreated control fish survived, and a sample of the remaining fish was heavily infested with I. necator. All remaining control fish were dead by 5 d posttreatment. The KMnO4 treatment significantly curtailed the initial mortality (survival = 92.4%) and slightly reduced the high parasite loads at 2 d posttreatment. However, fish mortalities increased dramatically over the next 3 d (survival at 5 d posttreatment = 37.5%), and parasite loads from sampled fish remained high. The CuSO4 treatment was effective in significantly lowering the parasite load (almost eliminating I. necator) and maintaining a high fish survival (87.5%) by 5 d posttreatment. The findings in this study clearly demonstrate that CuSO4 is a viable treatment for ichthyobodosis in tanks. PMID- 19306610 TI - Transmission of the parasite Ichthyophonus hoferi in cultured rainbow trout and comparison of epidemic models. AB - The epidemic process of the parasite Ichthyophonus hoferi in cultured rainbow trout Oncorhynchus mykiss was quantitatively estimated by both the cohabitation experiment and two standard models (the Kermarck-McKendrick model and the Reed Frost model). For analysis of the parasite transmission by cohabitation, fish in two replicate tanks were exposed to 1, 5, or 10 infected fish, and daily mortality was counted for 102 d. Despite simple experiments for artificial exposure to the pathogen, the daily estimate of dead fish in the Kermarck McKendrick model did not fit the observed number of dead fish in the experiment. In contrast, when the longest possible incubation period (generation time) was assumed to be 51 d in the Reed-Frost model, the estimated number of dead fish in discrete generations was close to the observed number of dead fish. If the time unit was 51 d, the estimated mortalities in the generation-based Kermarck McKendrick model were significantly correlated with observed mortalities. These results suggest that the deterministic aspects of the epidemic process of the parasite can be quantitatively demonstrated on a 51-d timescale or longer, whereas transmission on a daily timescale is uncertain. PMID- 19306611 TI - Genetic analysis of paramyxovirus isolates from Pacific salmon reveals two independently co-circulating lineages. AB - Viruses with the morphological and biochemical characteristics of the family Paramyxoviridae (paramyxoviruses) have been isolated from adult salmon returning to rivers along the Pacific coast of North America since 1982. These Pacific salmon paramyxoviruses (PSPV), which have mainly been isolated from Chinook salmon Oncorhynchus tshawytscha, grow slowly in established fish cell lines and have not been associated with disease. Genetic analysis of a 505-base-pair region of the polymerase gene from 47 PSPV isolates produced 17 nucleotide sequence types that could be grouped into two major sublineages, designated A and B. The two independently co-circulating sublineages differed by 12.1-13.9% at the nucleotide level but by only 1.2% at the amino acid level. Isolates of PSPV from adult Pacific salmon returning to rivers from Alaska to California over a 25-year period showed little evidence of geographic or temporal grouping. Phylogenetic analyses revealed that these paramyxoviruses of Pacific salmon were most closely related to the Atlantic salmon paramyxovirus (ASPV) from Norway, having a maximum nucleotide diversity of 26.1% and an amino acid diversity of 19.0%. When compared with homologous sequences of other paramyxoviruses, PSPV and ASPV were sufficiently distinct to suggest that they are not clearly members of any of the established genera in the family Paramyxoviridae. In the course of this study, a polymerase chain reaction assay was developed that can be used for confirmatory identification of PSPV. PMID- 19306612 TI - Decreased mortality of Lake Michigan Chinook salmon after bacterial kidney disease challenge: evidence for pathogen-driven selection? AB - In the late 1960s, Chinook salmon Oncorhynchus tshawytscha from the Green River, Washington, were successfully introduced into Lake Michigan. During spring from 1988 to 1992, large fish die-offs affecting Chinook salmon occurred in the lake. Multiple ecological factors probably contributed to the severity of the fish kills, but the only disease agent found regularly was Renibacterium salmoninarum, the causative agent of bacterial kidney disease. In this study, survival after challenge by R. salmoninarum was compared between two Chinook salmon stocks: a Lake Michigan stock from Wisconsin (WI) and the progenitor stock from the Green River. We found that the WI stock had significantly greater survival than the Green River stock. Next, the WI and Green River stocks were exposed to the marine pathogen Listonella anguillarum (formerly Vibrio anguillarum), one of the causative agents of vibriosis; survival after this challenge was significantly poorer for the WI stock than for the Green River stock. A close genetic relationship between the Green River and WI stocks was confirmed by analyzing 13 microsatellite loci. These results collectively suggest that disease susceptibility of Lake Michigan Chinook salmon has diverged from that of the source population, possibly in response to pathogen-driven selection. PMID- 19306613 TI - Induction and evaluation of proliferative gill disease in channel catfish fingerlings. AB - Proliferative gill disease (PGD) was first reported in channel catfish Ictalurus punctatus at commercial farms in 1981 and is caused by the myxozoan parasite Henneguya ictaluri. The disease affects the gills and is characterized by severe branchial inflammation, epithelial hyperplasia, lamellar fusion, and lysis of chondrocytes. Presumptive diagnosis is based on the presence of lytic areas in the cartilage of the primary lamellae on microscopic examination and is confirmed histologically by the presence of the organism. In these trials, PGD was induced by exposing channel catfish fingerlings to fresh or aged infectious water collected from a pond containing fish diagnosed with severe PGD. The severity of disease was graded by histological scoring and microscopic examination of wet mounts to determine the percentage of gill filaments containing chondrolytic lesions. Exposure of fish to infectious pond water was shown to produce pathological lesions consistent with PGD, and the percentage of gill filaments containing chondrolytic lesions was positively correlated with histological scoring of gill pathology. The number of trophozoite stages in the gills was shown to increase with the severity of the disease. In most cases, however, parasitic cells were not observed in tissue samples with chondrolytic lesions during the early stages of infection. These observations indicate that pathology and lysis of chondrocytes can occur prior to detection of the organism by histopathology. Exposing fish to infectious pond water that was aged for 1 d produced negligible gill pathology and implies that the infectivity of the H. ictaluri actinospore stage is short lived. Removing fish from the source of infection promoted repair of damaged gill tissue; within 14 d of fish transfer to clean water, gill pathology associated with the acute infection was negligible. PMID- 19306614 TI - Comparison of in vitro antimicrobial susceptibility in Flavobacterium psychrophilum isolated from rainbow trout fry. AB - The aim of this study was to demonstrate the presence of Flavobacterium psychrophilum in the west Aegean region of Turkey and to evaluate the in vitro susceptibility of F. psychrophilum (isolated from the fry of rainbow trout Oncorhynchus mykiss) to seven antimicrobial agents, as determined by the disk diffusion and agar dilution methods. A total of 250 rainbow trout fry (weight = 2 5 g; total length = 3-6 cm) were examined, and 20 bacterial isolates were phenotypically identified. Antimicrobial agents included in this investigation were amoxicillin-clavulanic acid (AMC), erythromycin (E), enrofloxacin (ENR), florfenicol (FFC), gentamicin (CN), oxytetracycline (OT), and sulfamethoxazole trimethoprim (SXT). Disk diffusion and agar dilution methods were performed according to published standards. Minimum inhibitory concentration (MIC) ranges were determined using the agar dilution method for F. psychrophilum isolates. Resistance of F. psychrophilum to CN (disk diffusion method: 70%; agar dilution method: 95%), E (65%; 100%), and SXT (75%; 100%) was high using both methods. Resistance to ENR (10%; 15%) and FFC (25%; 25%) was low with both methods; MIC90 (minimum concentration required to inhibit bacterial growth by 90%) was 4 microg/mL for ENR and 16 microg/mL for FFC. Ninety percent of the F. psychrophilum isolates were resistant to AMC based on the disk diffusion method, while only 15% of isolates showed resistance based on the agar dilution method. For OT, 20% of isolates were resistant based on disk diffusion, while 75% exhibited resistance based on agar dilution. The importance of susceptibility testing when facing an outbreak of F. psychrophilum at a fish farm is obvious; however, the discrepancies between testing methods for AMC and OT require further studies. PMID- 19306615 TI - Addiction and the pharmacology of cannabis: implications for medicine and the law. AB - The topic of drug addiction or misuse of drugs has numerous far-reaching ramifications into areas such as neuroscience, medicine and therapeutics, toxicology, epidemiology, national and international economics and politics, and the law. The general principles of drug addiction are first summarised. A recurring and intrinsic problem is lack of adequate characterisation of the independent variable, namely the drug taken. Secondly, it is not feasible to allocate subjects randomly to treatments. Thirdly, the heterogeneity of different forms of addiction precludes facile generalisations. "A problem drug user is anyone who experiences social, psychological, physical, or legal problems related to intoxication, and/or regular excessive consumption, and/or dependence as a consequence of their use of drugs" (UK Advisory Council on Misuse of Drugs, 1982). Cannabis is a genus of flowering plants whose products are used as recreational drugs. Claims have been made for a range of therapeutic properties. Its two main active principles are delta9 - tetrahydrocannabinol (THC) and cannabidiol (CBD). These compounds have contrasting pharmacological properties. THC is suspected of causing psychotic phenomena, but CBD seems more sedative and may even be antipsychotic. The past use of cannabis, particularly the concentrations of THC and CBD, can be monitored with hair analysis. Recent studies involving the administration of THC and CBD to human subjects are reviewed. Suggestions are made for further research into the pharmacology and toxicology of CBD. Such data may also point to a more rational evidence-based approach to the legal control of cannabis preparations. PMID- 19306616 TI - Cause of death in judicial hanging: a review and case study. AB - The cause of death in judicial hanging is controversial and often attributed to 'hangman's fracture' of the second cervical vertebra. Research has shown that such fractures are the exception in judicial hangings and the cause of death can be attributed to a range of head and neck injuries, particularly compression or rupture of the vertebral and carotid arteries leading to cerebral ischaemia. The rapidity of loss of consciousness and death is highly dependent upon knot positioning and the length of drop which has varied through the history of hanging as a capital punishment in the UK. The skeletal remains of Mr. George Kelly, wrongfully hanged for murder at Walton prison, Liverpool (1950) were exhumed, examined and are reported on herein. The first cervical vertebra was found to be fractured but no 'hangman's fracture' of the axis--second cervical vertebra--was present. The hangman (Mr. Albert Pierrepoint) has been quoted as stating that the hanging of Mr Kelly took longer than 'it should have' (Dernley and Newman, 1989) but no skeletal evidence of death by strangulation was found by the authors. Unconsciousness, if not death, would probably have been rapid due to vertebral artery damage as a result of the observed neck fracture, although this cannot be concluded with absolute certainty. PMID- 19306617 TI - Coincident deaths: double suicide or murder-suicide? AB - A retrospective review was undertaken in South Australia over an 11-year period from July 1996 to June 2007 of all multiple, unnatural, non-accidental deaths in adults (> 16 years) looking for cases of double suicide and murder-suicide. All cases had undergone a full police and coronial investigation and full autopsy. There were seven cases of double suicide involving 14 individuals: nine males and five females ranging in age from 29 to 79 years (average 49 yrs). Over the same period there were eight murder-suicides involving 16 individuals: nine males and seven females ranging in age from 17 to 83 years (average 49 yrs). All the murder suicide perpetrators were male and the majority of the victims were female (7/8). All eight perpetrators committed suicide with gunshot wounds to the head (N = 8; 100%); one victim was manually strangled and the remainder died from gunshot wounds to the head, neck and upper chest. In contrast, all but two of the double suicide victims died of carbon monoxide toxicity, with the remaining couple dying from mixed drug toxicity. While differentiating between these two groups may be difficult, double suicide victims used much less violent methods than perpetrators of murder-suicide and often had histories of chronic debilitating disease. Spousal and familial links characterised both groups. PMID- 19306618 TI - Who are sexual offenders? A survey of pre-trial psychiatric reports. AB - Sexual offending is a serious and growing problem in our society. The aim of the study was to investigate the main characteristics of people charged with sexual offences who presented before the criminal courts. The survey was conducted retrospectively between August 2001 and August 2006 on pre-trial court reports stored in a computer database shared by forensic psychiatrists using The Grange consulting rooms in West Yorkshire. A data collection form was used to gather the characteristics of sexual offenders. The data collected was analysed using descriptive statistics. Our survey revealed the following results. Out of 78 cases evaluated, the commonest sexual offence was against children (68.8%). Thirty-two per cent of those with paedophilic behaviour had a history of childhood sexual abuse. Rape was alleged as the main sexual offence in 27.27% cases. Substance abuse (30.76%) and sexual motivation (42.30%) were the predominant motives for offending behaviour. Low rates of sexual fantasy and sadistic behaviour (8.97%) in our sample could be due to the non-disclosure by sexual offenders. Mental disorder was observed in 7.69% cases. Significant personality factors were observed in 14.10% of the sample. A sexual offending treatment programme was recommended in 57.69% cases. A very high risk of re offending was recorded in 32.25% cases. Of the total sample, 93.50% were deemed fit to plead. PMID- 19306619 TI - Aftercare under Section 117 of the Mental Health Act 1983: awareness and practice. AB - The aim of this study is to investigate the level of awareness amongst consultants regarding Mental Health Act (MHA) Section 117 aftercare, their responsibilities and current practice. We conducted a cross-sectional survey alongside a retrospective study of the Trust Section 117 aftercare registers. Seventy-four per cent of the consultants were aware of the Section 117 aftercare registers and the need for review of aftercare. Ninety-four per cent were aware of the financial implications of Section 117. However, only 52% of the consultants regularly contributed to the review process to identify patients who were no longer in need of aftercare under Section 117. Over a five-year period, 150 patients were discharged from Section 117, of whom only 4% were officially discharged following review of the care plan. Health and social services have a legal obligation to provide aftercare to all eligible patients free of charge under Section 117. With the new Mental Health Act 2007 this will now include patients on Supervised Community Treatment Orders. It is important to have a clear understanding of Section 117 to ensure patients' needs are met and to allow effective use of resources. PMID- 19306620 TI - Deaths of homeless unclaimed persons in South Delhi (2001-2005): a retrospective review. AB - Homeless people are at increased risk of death for several reasons. We reviewed 735 cases of homeless, unclaimed persons found dead in public places in South Delhi who were brought for autopsy at the Department of Forensic Medicine, All India Institute of Medical Sciences, New Delhi, during the study period from 2001 2005. Most of the deceased were male (n = 645, 87.75%) with a M: F ratio of 7.17:1. The commonest age group involved was 31-40 years (n = 228, 31.02%). Most victims (n = 451, 61.36%) died from natural causes, followed by accidental deaths (n = 233, 31.70%) and homicides (n = 26, 3.54%). The majority of deaths were reported in the rainy season (n = 284, 38.64%). The deceased were mostly recovered from the road or road side (n = 352, 47.89%). Few studies on this subject have been published in India. More research is needed to identify the health-related problems of such people and possible contributory factors to mortality. PMID- 19306621 TI - Pathology of firearm mortalities in the Niger Delta region of Nigeria: a study of 136 consecutive autopsies. AB - A prospective autopsy study of firearms-related death was carried out by the authors in the Niger Delta region of Nigeria. The purpose of the study was to evaluate the pattern, frequency, and anatomical sites of wounds, also the cause of death and the age and sex distribution of victims. The autopsies were performed after being served with the coroner's inquest forms. In all cases standard autopsy procedures were adopted and reports issued. A total of 136 consecutive firearm deaths were autopsied. There were 122 (89.7%) males and 14 (10.3%) females, giving a male to female ratio of 8.7:1. The highest number of deaths, 95 (69.8%), occurred in the age group of 10-39 years with a peak of 51 (37.5%) deaths at 20-29 years. In 112 (82.4%) cases death occurred from homicidal gunshot wounds. The commonest targets of gunshot wounds were the head, 58 (42.6%); multiple anatomical sites, 30 (22.1%); chest, 16 (11.8%); abdomen, 11 (8.1%) and neck, 10 (7.4%). Haemorrhagic shock was the cause of death in 131 (96.3%) cases. Significant causes of traumatic death in the Niger Delta region of Nigeria are gunshot wounds and associated complications. Stringent laws on illegal firearms handling should be passed and enforced by the government to reduce the frequency of such deaths. PMID- 19306622 TI - Manufacturing fault causes a false negative for haematuria using Makromed m81 urinalysis test strips: a case report. AB - A 35-year-old male patient presented with painful gross macroscopic haematuria. Urinalysis using Makromed test strips showed a negative reading for blood. Subsequent laboratory analysis showed the presence of gross haematuria. A referral to a consultant urologist confirmed a diagnosis of renal stones. Subsequent investigations by the author indicated that a manufacturing fault was responsible for this error. Discussion with the manufacturer identified a fault in the packaging which led to photosensitive light degradation of enzyme pathways, resulting in a false negative finding. In situations when gross clinical findings are not supported by on -site diagnostic tests, the clinician should consider the possibility that the diagnostic tool is at fault and not the clinician. Makromed urine tests strips (MUTS) have been used in this practice for several years. There had been no previous concerns raised by any clinician within the group practice, nor by the Primary Care Trust, or any authoritative professional body. Argyropoulos et al. (2004) reported that renal stones cause haematuria in 92.9% of cases. PMID- 19306623 TI - Avulsion of the left internal mammary artery graft after minimally invasive coronary surgery: fatal complication or medical error? A case report. AB - Minimally invasive direct coronary artery bypass (MIDCAB) is performed through a left anterior mini-thoracotomy without the use of a cardiopulmonary bypass and offers greater potential for more rapid recovery, reduced pain and a decreased need for blood transfusion than conventional coronary artery bypass grafting. Few major complications of the MIDCAB procedure have been reported in the literature since the first intervention was performed in 1995, but the most serious one is avulsion of the left internal mammary artery (LIMA) graft near the site of anastomosis with the left anterior descending coronary artery. Forensic issues regarding the role of the surgeon in causing this life-threatening emergency condition have not been discussed. We report here the case of a 48-year-old man who died 18 days after a MIDCAB of massive thoracic bleeding due to the avulsion of the LIMA graft. We discuss the probable etiopathogenesis of this fatal complication from a forensic point of view. PMID- 19306624 TI - Acute intoxication by triazolam and promazine: a case report. AB - A fatality due to ingestion of triazolam and promazine is reported. Triazolam is a benzodiazepine widely prescribed as a hypnotic drug for the treatment of sleep disorders. Promazine is a neuroleptic drug. There is no previous evidence in the literature of death due to an overdose related to the contemporaneous intake of these two drugs. In this report the authors present the case of a 76-year-old woman who was found deceased at home with no evidence of trauma or asphyxia; near the body several empty pharmaceutical boxes containing triazolam and promazine were noticed. Toxicological analyses were performed and drug levels measured by means of gas chromatography coupled with mass spectrometry. The triazolam concentration in each specimen was as follows: blood 1100ng/ml; gastric content 1300ng/ml; the promazine concentration in blood and in gastric content was 3450ng/ml and 5800ng/ ml respectively. Based on the autopsy findings, patient history and toxicological results, the cause of death was determined to be acute intoxication due to the effect of triazolam and promazine and the manner of suicide. PMID- 19306625 TI - Appointed bullet--a premeditated murder followed by suicide: a case report. AB - A 21-year-old girl was fatally injured by her 25-year-old brother-in-law, who was deeply in love with her. She was shot in the abdomen with a locally-made pistol. The man then shot himself in the front of his head with the same gun. He died on the spot while the girl, who was seriously injured, died in hospital two days later. Police recovered two empty cartridges from the alleged site that had the names of the boy and girl written on them in blue ink. Although shooting is the most common method of homicide and subsequent suicide, to the authors' knowledge the case described here is unique and has not been reported elsewhere in the literature. PMID- 19306626 TI - [Current understanding of new standards of patient care in cardiovascular anesthesia: preface and comments]. AB - The search to define new standards of patient care is of particular relevance for the anesthesiologists providing cardiovascular anesthesia. Because of conflicting results on multiple issues, however, it is often difficult to understand new standards of patient care especially for occasional cardiovascular anesthesiologists. The objective of this series of articles is to summarize evidence of clinical and basic research and introduce current understanding of new standards of patient care in cardiovascular anesthesia. The following topics are discussed in each article: use of cardiovascular drugs for cardiac surgery, myocardial preconditioning in anesthesia, hemostatic management for cardiac surgical patients, blood glucose control during cardiac surgery, cognitive dysfunction after cardiac surgery, spinal protection in aortic surgery, ventricular assist device and heart transplantation in Japan. Further investigations in this area are critical in order to provide optimal patient care. PMID- 19306627 TI - [Use of cardiovascular drugs for cardiac surgery--based on recent studies]. AB - Protective effects of each cardiovascular drug on major organ functions are reviewed. Based on these effects, proper use of the drugs is presented. It is probable that norepinephrine and dobutamine have the most protective effect against major organ dysfunctions. It has been shown that perioperative beta blocker administration reduces cardiovascular complications. Nicorandil is likely to have ischemic preconditioning properties. In case of hypotension, low doses of noradrenaline should be administered first, and then dobutamine should be added. In case of systolic dysfunction, low doses of dobutamine should be chosen first, and then noradrenaline, and finally olprinone could be added. In case of hypotension and systolic dysfunction, combination of norepinephrine and dobutamine is the first choice. Then adrenaline could be added. When ischemic heart disease exists, nicorandil should be given. When decreasing or stabilizing heart rate is required, an ultra-short acting beta-blocker, such as landiolol, is recommended. To maintain hypotension in a certain situation, prostaglandin E1 is better to use than nicardipine, diltiazem, and nitroglycerin. It is important to administer the drugs focusing on postoperative complications and outcome. PMID- 19306628 TI - [Myocardial preconditioning in anesthesia: from bench to bedside]. AB - Myocardial ischemic preconditioning is a procedure giving powerful protection against myocardial ischemia and infarction. Clinical application of this property to anesthesia might improve anesthetic management of patients with cardiac diseases. Volatile anesthetics, including sevoflurane and isoflurane, can protect ischemic myocardium by activation of ATP-sensitive K channels (K(ATP)) and several clinical trials show that volatile anesthetics can prevent myocardial damage during operations compared with propofol, a popular intravenous anesthetic. Nicorandil is a potent K(ATP) opener and clinical application of nicorandil also can reduce myocardial damage during anesthetic management of patients undergoing off-pump CABG and patients with cardiac risks undergoing non cardiac operations. In addition, diabetes mellitus and hyperglycemia may abolish the protective effect of preconditioning by impairing K(ATP). From point of view of clinical application of preconditioning, anesthetic maintenance with volatile anesthetics instead of intravenous anesthetics, such as propofol, and administration of nicorandil throughout perioperative period, is recommended to patients with heart diseases and control of blood glucose is useful to maintain the protective effect of preconditioning. PMID- 19306629 TI - [Hemostatic management for cardiac surgical patients in the post-aprotinin era]. AB - Antifibrinolytic therapy is effective in reducing postoperative bleeding and allogeneic transfusion in patients undergoing cardiac surgery with cardiopulmonary bypass. Aprotinin is a bovine serine protease inhibitor which potently inhibits plasmin, and two lysine analogues, epsilon-aminocaproic acid and tranexamic acid, inhibit the activation of plasmin by binding to plasminogen. Although aprotinin has been considered more efficacious than lysine analogues in reducing blood loss, recent clinical studies in cardiac surgical patients suggested increased mortality in aprotinin compared to lysine analogues. Why could aprotinin be more harmful to high-risk cardiac surgical patients? Are there enough safety data on the use of epsilon -aminocaproic acid and tranexamic acid? Potential harms of inhibiting endogenous fibrinolytic system have not been systematically reviewed. This review article presents a current perspective on the efficacy and safety of antifibrinolytic agents based on the available basic science and clinical data. Further, the hemostatic strategies for complex cardiac surgical patients will be proposed. PMID- 19306630 TI - [Blood glucose control during cardiac surgery]. AB - Since the strict blood glucose control by intensive insulin therapy was introduced as a life-saving maneuver, a lot of beneficial effects have been reported. The intensive insulin therapy has also been recommended in cardiac surgeries, particularly with cardiopulmonary bypass (CPB) which often induces hyperglycemia resulting from the excessive stress response. Recently, however, some groups have reported that intensive insulin therapy often leads to a serious complication like hypoglycemia, and they have expressed opposition to the therapeutic benefit. These studies indicated that the same target level of blood glucose did not provide the beneficial outcome to all patients, and that the optimal level of blood glucose differed depending on the individual patient and care setting. In order to determine the target level for blood glucose resulting in the lowest risk-to-benefit ratio, we must take into account various factors, such as variability of blood glucose concentration, presence or absence of diabetes, care setting, and perioperative nutritional management. PMID- 19306631 TI - [Cognitive dysfunction after cardiac surgery]. AB - The advancement of surgical and anesthetic techniques reduced cardiac morbidity and mortality. As a result, the attention has been focused on the perioperative cerebral complication, which contribute to more than 20% of perioperative deaths. Cognitive dysfunction is associated with increased medical costs, decreaged quality of life, and mortality. However, the incidence of cognitive dysfunction has a wide variety due to methodological variations in evaluating cognitive dysfunction, showing the complexity of cognitive dysfunction. The score of cognitive test can include learning effect and natural decline of cognitive dysfunction in elderly population. The research including the control group seems to overcome these issues, but still remains to have problems. Further, patients with cardiac disease have comorbid factors to induce postoperative sequelae compared to healthy population. We describe these issues which require continued researches. PMID- 19306632 TI - [Spinal protection and aortic surgery]. AB - Motor dysfunction following thoracoabdominal aortic surgery remains as one of their devastating postoperative complications. Recently, many studies have provided information for the mechanisms of ischemic spinal cord injuries and for some strategies against spinal cord ischemia in the perioperative period. Especially, advances in stimulation technique using multipulse could make intraoperative monitoring of functional integrity of motor pathways possible by recording myogenic motor evoked potentials. We summarize the strategies for spinal cord protection, including spinal cord functional monitoring and anesthetic techniques during thoracoabdominal aortic surgery. PMID- 19306633 TI - [Ventricular assist device and heart transplantation in Japan]. AB - Ventricular assist device (VAD) has been applied for treatment of end-stage heart failure. Three major purposes of VAD are a bridge to transplantation, a bridge to recovery and a destination therapy (permanent VAD implantation). In Japan, only paracorporeal ventricular assist device (Toyobo-NCVC type) is approved for coverage by health insurance. Survival after Toyobo VAD implantation has been improving greatly Survivals 1, 3, 5-years after implantation are 75%, 75% and 58% respectively in our experience. Several implantable VADs are under clinical trials in Japan. Implantable VAD has many advantages, such as allowing patients to be discharged and being resistant to infection. Heart transplantation was performed in more than 70,000 cases worldwide. At present, approximately 3,000 transplantations were performed annually. Survivals after 1, 3, 5, 10-years are 86%, 79%, 72% and 51%. Fifty-seven heart transplantations were performed in Japan since 1999. Survivals after 1, 3, 5-years in Japan are 98%, 98% and 94%, which are far superior to world average results. PMID- 19306634 TI - [Anesthesia induction for patients with cervical spinal disease]. AB - BACKGROUND: In the cases in which the flexibility of the patient's neck is limited, it is often difficult to achieve intubation. Fiberoptic intubation has been chosen for patients with arthroses. However, as it requires techniques, it has been difficult for inexperienced anesthesiologists. METHODS: Awake induction using AirWay Scope was performed for 8 patients with cervical spinal diseases. Midazolam, propofol or dexmedetomidine is selected as a sedative drug. RESULTS: Intubation was performed easily and safely without any complications in all cases. No patients had memories of intubation. CONCLUSIONS: Awake intubation using AirWay Scope for patients with cervical spinal diseases is a safe and useful anesthesia method. As dexmedetomidine causes no respiratory depression and can expect cooperation from patients, it may give safe and efficient sedation in awake intubation cases. PMID- 19306635 TI - [Influence of endotracheal tube cuff lubrication on postoperative sore throat and hoarseness]. AB - BACKGROUND: Sore throat and hoarseness are common postoperative complications in patients who undergo tracheal intubation. In this study, we evaluated the severity of postoperative sore throat and the incidence of hoarseness in 60 patients after tracheal intubation. METHODS: 60 patients (ASA PS 1 or 2, 29 males and 31 females) scheduled for general anesthesia with endotracheal intubation were enrolled in this study. They were divided into three groups. Lidocaine 4% was sprayed into the trachea in the sprayed group (n=20). The distal end of the endotracheal tube was lubricated with 2% lidocaine jelly in the lubricated group (n=20). No intervention to the endotracheal tube was carried out in the no intervention group (n=20). We evaluated the visual analogue scale (VAS) of sore throat and the incidence of hoarseness in each group at the end of general anesthesia and the next day. RESULTS: VAS scores of sore throat at the end of anesthesia were 9.2 +/- 3.4 mm in the sprayed group, 27.8 +/- 5.7 mm in the lubricated group, and 11.8 +/- 4.4 mm in the no-intervention group. VAS scores on the next day were 2.5 +/- 1.4 mm in the sprayed group, 14.0 +/- 4.3 mm in the lubricated group, and 2.2 +/- 1.7 mm in the no-intervention group. Both VAS scores at the end of anesthesia and the day after anesthesia were significantly higher in the lubricated group than others (P<0.05). However, there was no significant difference in hoarseness among the three groups. CONCLUSIONS: In this study, VAS scores at the end of anesthesia and the next day were both significantly higher in the lubricated group than in others. Furthermore, there is no significant difference in VAS between the sprayed group and the no intervention group. These data suggest that lidocaine jelly lubrication to the endotracheal tube reinforces the severity of sore throat. On the other hand, there was no significant difference in VAS between the sprayed group and the no intervention group. This suggests that lidocaine sprayed to the trachea does not reduce postoperative sore throat. PMID- 19306636 TI - [Efficacy and safety of AirWay Scope in using a bronchofiberscope in patients with difficult airway]. AB - Case 1: A 41-year-old woman was scheduled for pinning of fractured finger and repair of a ligamentum. Preoperative problems did not exist except mandibular hypoplasia. General anesthesia was induced and Cormack classification was grade III in laryngeal view by laryngoscope and we tried a new videolaryngoscope PENTAX AirWay Scope (AWS). We did not catch the epiglottis directly by Intlock and the target mark was not located at the middle of the vocal cord. Bronchofiberscope was guided to the vocal cord through a tracheal tube attached to AWS by another anesthesiologist and the patient was successfully intubated. Case 2: A 46-year old man was scheduled for anterior screw-plate fixation due to C3/4 herniated disc. AWS using bronchofiberscope procedure was tried to keep the neutral position of the neck. The patient was successfully intubated. We did not view the bronchofiberscope and used it just as a flexible bougie. Case 3: A 56-year-old man was scheduled for free radial forearm flap reconstruction of a diabetic necrotic toe. Ten minutes were needed to finish intubation due to difficult airway (Cormack classification grade III) when free latissimus dorsi flap reconstruction was perfomed two month before. Hence, AWS using a bronchofiberscope procedure was tried first and 39 seconds were needed to intubate. We concluded that AWS using bronchofiberscope procedure was useful for patients with difficult airway, though two anesthesiologist, one keeping AWS and the other using a bronchofiberscope, were needed. PMID- 19306637 TI - [Acupuncture treatment for a patient with prolonged consciousness disturbance after a brain tumor surgery]. AB - Acupuncture has long been applied as a therapeutic technique in China, Japan, Korea and other countries. Recently, its application began to be extended to the treatment of neural disorders. We experienced a 13-year-old boy with prolonged consciousness disturbance after a pineal tumor surgery and muscle contracture of lower extremity by long-term recumbency. We applied him acupuncture treatment for 4 months which was effective to alleviate these symptoms. Repeated IMP SPECT showed improvement of the cerebral blood flow (CBF) during the course of acupuncture therapy. Acupuncture was effective to improve prolonged coma after a brain surgery and also muscle contracture by long-lasting recumbency. CBF showed a slight increase along with the recovery of consciousness suggesting a strong relevance between CBF and improvement of these symptoms. PMID- 19306638 TI - [Central venous catheterization complication by a guide wire]. AB - Central venous catheterization using the Seldinger technique is a well known and often used method. On the other hand, there are also well known complications by needle puncture or by indwelling catheter, there are few reports about a guide wire which got hung up around the tricuspid valve. We report a case in which a guide wire got hung up to the chordae tendineae of the tricuspid valve. To insert the AVA 3Xi (Edwards life science Co. Iervine) from the right internal jugular vein, we inserted a guide wire without ease. Resistance appeared when we tried to remove the wire for 20 cm from the inserted state. The X-ray and the transesophageal echocardiography, showed the guide wire in the right ventricle. As actions to be taken, we advanced the central vein catheter of the EXCV catheter kit (Nippon Sherwood Medical Industries Co., Ltd.) to the tip, and a the guide wire was easily removed. There are many reports of the complication by the central venepuncture, but there are few reports about the guide wire which was entrapped in the vicinity of a tricuspid valve. The tip of the guide wire in this case was bent excessively, but the cause of the damage did not become clear by investigation. When a guide wire became hard to withdraw, we should never withdraw a guide wire blindly, but should search a cause and we should use the material which was matched with the cause. PMID- 19306639 TI - [Sedation during day surgery for oral implantation]. AB - BACKGROUND: Oral implantation surgery is performed under sedation without monitoring consciousness, but BIS monitor is useful for evaluation of sedation level. Therefore we investigated whether BIS monitor is useful for this surgery. METHODS: Sixty-four patients scheduled for oral implant surgery were analyzed retrospectively. Patients were classified into two groups; BIS monitor was used (Group B) or not (Group A). We investigated the dose of midazolam necessary for intraoperative sedation, and for patient's satisfaction. RESULTS: The dose of midazolam in Group B was greater than that in Group A. The dose in Group B was higher than that in Group A according to patient's satisfaction. CONCLUSIONS: BIS monitor is useful for oral implant surgery. PMID- 19306640 TI - [Anesthetic management of a patient with severe ovarian hyperstimulation syndrome]. AB - We report anesthetic management of a patient with severe ovarian hyperstimulation syndrome (OHSS). A 33-year-old woman presented with severe abdominal acites and effusion after ovarian stimulation with HMG followed by ovulation induction with HCG. She was suspected of having intraabdominal hemorrhage because of ectopic pregnancy. Emergency laparotomy was performed under general anesthesia. Massive ascites and intraabdominal hemorrhage were observed and patient was treated by adequate fluid infusion and blood transfusion. Patient was treated with non invasive positive pressure ventilation (NPPV) after operation and recovered. It is essential to maintain a correct balance of fluids, through appropriate intravenous infusion of crystalloids, albumin, blood transfusion and an adequate urine output. Further it is important to prevent thromboembolic event. Knowledge of the clinical features, complications and acute management of OHSS will permit the anesthesiologist to treat these patients in an optimal fashion. PMID- 19306641 TI - [Report of an inquiry about epidural anesthesia for abdominal aortic aneurysm surgery in Japan]. AB - BACKGROUND: Recently there are growing number of patients suffering from the abdominal aortic aneurysm (AAA), and we have many occasions to anesthetize these patients under coagulation therapy as well. Moreover, the risk of epidural hematoma increases when the operation of the AAA is performed with epidural technique because the operation is usually done with perioperative heparinization. For these reasons, we investigated the current situations of clinical practice in Japan in terms of the epidural anesthesia for AAA surgeries. METHODS: The questionnaires were sent to all 998 Japanese Society of Anesthesiologists certified training hospitals in October 2005, anonymously asking about current practices concerning AAA cases, anesthetic managements, use of epidural anesthesia and the experience of any complications. Fifty one per cent of the total questionnaires were returned and 94% of them could be analyzed. Seventeen per cent of responders were from university institutes or university related hospitals and 75% of them were from general hospitals of secondary critical care centers. RESULTS: This survey showed that the operations of AAA were done at 308 (64% of the responders) hospitals and the total number of AAA surgeries carried out was estimated to be 6,321 the last year. This figure was very close to the number announced by Japan Society of Cardiothoracic Surgeons. The epidural anesthesia was routinely used for AAA surgeries with general anesthesia in 224 hospitals (64%), but it was not used in 51 hospitals (14%). The standard protocol for the application of epidural anesthesia had been installed in 56% of hospitals and mostly in 187 hospitals (as several answers can be chosen). The epidural catheters were inserted the day before operation. There were 113 hospitals in which less than 2 hours was required from epidural puncture to heparinization, but in 141 hospitals it took more than 12 hours. However, on the contrary, many anesthetists answered that the risk of the epidural hematoma had been the cause of not routinely using the epidural anesthesia in AAA operations. We had 30 hospitals in which epidural hematoma had occurred in the cases not limited to AAA surgery and 17 cases of them resulted in severe complications afterwards. CONCLUSIONS: This investigation clarified the current situations of the clinical practice in Japan of the use of epidural anesthesia for AAA surgeries. PMID- 19306642 TI - [The incidence of critical arrhythmia related to epinephrine under halogenated inhalational anesthesia--report of the Subcommittee on Drugs, Committee on Patient Safety and Risk Management, and Committee on Administrative Affairs Japanese Society of Anesthesiologists]. AB - The incidences of intra-operative critical arrhythmia related to epinephrine under halogenated inhalational anesthesia were analysed according to questionnaire to 1108 JSA (Japanese Society of Anesthesiologists) Certified Training Hospital. The survey details included prospective (from July 16th, 2008 to Aug 15th, 2008) and retrospective (from Jan 1st, 2007 to Dec 31th, 2007) incidences of critical arrhythmia due to epinephrine under halogenated inhalational anesthesia. Among the 1108 institutions, effective responses were obtained from 583 institutions. A total of 1.2 case per 100,000 cases of critical arrhythmia were recorded in the retrospective study, and no case was recorded in the prospective study. The use of epinephrine under halogenated inhalational anesthesia was safe, but careful use is recommended. PMID- 19306643 TI - [Endoscopic endonasal surgery for extrasellar tumors: case presentation and its future perspective]. AB - BACKGROUND AND OBJECTIVES: Endoscopic endonasal transsphenoidal surgery has been performed because of its advantages such as less invasive surgical management and more aggressive tumor removal of extrasellar lesions. In 2003, we began endoscope assisting surgery. In 2006, we completely switched to the endoscopic endonasal approach without microscope or nasal specula. Today, we report endoscopic pituitary and skull base surgery in our institute. The endonasal approach via the sphenoid ostium was carried out without nasal specula. Postoperative nasal packing was basically not needed in such cases. In cases with meningiomas, craniopharyngiomas and giant pituitary adenomas, which needed intra-dural procedure, nasal procedures such as middle nasal conchotomy, posterior ethmoidectomy and skull base techniques such as optic canal decompression and removal of the planum sphenoidale were carried out to gain the wider operative field toward anterior skull base and lower clivus. Navigation and US-Doppler were essential. Angled endoscope attained more successful removal of tumor under direct visualization extending into the cavernous sinus (GH secreting ademomas) and lower clivus (chordoma). In the case of CSF (cerebrospinal fluid) leakage during operation, a newly designed balloon catheter was placed in the sphenoid sinus to fix the abdominal fat and fibrin glue at the leakage point. In recent cases, dural opening has been sutured. In the combination of such techniques, a lumbar drainage system to prevent postoperative CSF rhinorrhea became needless in many cases. Angled suction tips, single-shaft coagulation tools and slim and longer holding forceps, all of which were newly designed for endoscopic surgery, were essential for smoother procedure. Endonasal endoscopic pituitary surgery has resulted in less invasive transsphenoidal surgery since no postoperative nasal packing is needed and there is less dependency on lumbar drainage. Although better techniques to prevent postoperative CSF leakage needs to be developed, this endoscopic pituitary surgery will become more common and will become a standard procedure. Endoscopic skull base surgery has enabled more aggressive removal of extrasellar tumors with the aid of nasal and skull base techniques. This endoscopic skull base surgery is more highly specialized, needs special techniques and surgical training. Selection of patients is also important. This also needs collaboration with ENT (ear, nose, throat) doctors. To be acknowledged as a safe and successful procedure in skull base surgery, this complex procedure may be preferably carried out only in center hospitals, which deal with many patients with a skull base lesion. PMID- 19306644 TI - [Detection of intraplaque hemorrhage with use of screening black-blood MRI]. AB - The evaluation of plaque characteritics as well as the degree of luminal narrowing is important in the management of carotid stenosis. Several reports have shown that intraplaque hemorrhage (IPH), which plays a pivotal role in plaque vulnerability, emits high signal intensity on T1-weighted images of high resolution black-blood (BB) MRIs. When an MRI is used to screen for vulnerable plaque, a short scanning duration and a simple MRI protocol are required. This study assessed the utility of screening BB-MRI to detect IPH. Ninety-three consecutive cases with atherosclerotic carotid stenosis scheduled for carotid endarterectomy (CEA) or carotid artery stenting between January 2005 and December 2007 were included. High-resolution BB-MRI scans were obtained with 2D double inversion recovery fast spin-echo sequences with cardiac gating. Screening BB MRIs were performed with 3D gradient-echo sequences suppressing blood flow signals by radiofrequency wave spatial saturation over the scanning field of 3D time-of-flight MRA. The relative overall plaque signal intensities with reference to the sternocleidomastoid muscle were calculated, and the correlation of intensities on T1-weighted images between high-resolution BB-MRI and screening BB MRI was investigated. We excluded 50 cases because of restenosis following CEA, use of a pacemaker, a long interval (>10 days) between high-resolution and screening BB-MRI examinations, or poor-quality images. Among the 43 cases included, the relative overall plaque signal intensity of high-resolution BB-MRI and screening BB-MRI were closely correlated (r = 0.568, p < 0.001). Screening BB MRI can be an alternative to high-resolution BB-MRI in the assessment of IPH and may be a useful tool for detection of vulnerable plaque. PMID- 19306645 TI - [Microvascular decompression of trigeminal neuralgia caused by vascular compression which penetrated the trigeminal nerve: a case report]. AB - A rare case of trigeminal neuralgia, caused by a branch of the superior cerebral artery running through the trigeminal nerve, was successfully treated by microvascular decompression. The nerve gap at the arterial penetration point was widened by the dissection of the epineurium to obtain sufficient mobilization of the artery and decompression of the nerve. This technique would be useful for securing neurovascular decompression, as well as for avoiding postoperative hypesthesia owing to the neurotomy described in previous reports. Preparing for such a rare case is important as preoperative magnetic resonance imaging (MRI) often fails to disclose precise diagnoses. PMID- 19306646 TI - [Neuroendoscopic management of a colloid cyst at the third ventricle using the high-definition flexible neuroendoscopic system: report of two cases]. AB - Colloid cysts are benign intracranial lesions located at the anterior roof of the third ventricle, which can result in the obstruction of the foramen of Monro and subsequent hydrocephalus or sudden death. Colloid cysts have been treated by microsurgical resection or by stereotactic aspiration. More recently, the use of rigid neuroendoscopy for colloid cysts has been attempted, but the efficacy of the flexible neuroendoscopic system is undetermined except for limited reports. A 40-year-old man and a 52-year-old man were presented with obstructive hydrocephalus owing to colloid cysts in the third ventricle, who were diagnosed by the typical findings of pre-operative CT scan and MRI. In the first case, endoscopic resection of the cyst was attempted using a high-definition flexible neuroendoscopic system with a CCD camera at the tip of the endoscope. The procedure relieved his symptoms. Hydrocephalus was improved after surgery, and postoperative MRI revealed the disappearance of the third ventricle lesion. Histological examination confirmed the diagnosis of colloid cyst. In the second case, endoscopic removal of the cyst wall and the evacuation of the cyst content were attempted using a high-definition flexible endoscopic system which resulted in the improvement of hydrocephalus and marked shrinkage of the third ventricle lesion observed in postoperative MRI. There was no recurrence in either patient. We recommend endoscopic treatment for colloid cysts using the high-definition neuroendoscopic system, which enables safe and thorough manipulation of the lesion and surrounding structures. PMID- 19306647 TI - [Preoperative evaluation of the aneurismal wall by using high-resolution MRI: report of 3 cases]. AB - Surgical treatment of large cerebral aneurysms may depend on preoperative information regarding the wall thickness and the intraaneurysmal thrombosis. When the neck of an aneurysm is thick due to atherosclerosis or there is a marked thrombosis inside the aneurysm, clipping may not be possible. Here we report three cases with large internal carotid artery aneurysms that were evaluated for the thickness of the aneurysm wall and the presence of thrombosis by 3 tesla magnetic resonance (MR) imaging. A high-resolution T2-weighted imaging (HRT2WI) modified for 3 tesla was used. In two cases, the images showed the thickened wall of the dome to some extent which apparently did not reach the neck. These aneurysms were successfully neck clipped. The third patient showed marked thickening of the wall, which may be associated with thrombosis, which extended to the neck. Intraoperative findings of the aneurysm wall were compatible with these factors, and removal of atherosclerotic plaque and thrombus from the neck and dome of the aneurysm was necessary before successful neck clipping. The HRT2WI may contribute to a better surgical strategy for large aneurysms which may have wall thickening. PMID- 19306648 TI - [Spontaneous regression of primary intracranial germinoma: a case report]. AB - Here we report a case in which an intracranial germinoma displayed spontaneous regression. An 11-year-old boy presented with polyuria and headache. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed tumors in the suprasellar and pineal regions, and obstructive hydrocephalus. As repeat MRI demonstrated shrinkage of these tumors, resection was deferred. The patient was discharged and followed up with serial MRI. The tumor continued to regress for three weeks; however, the patient was readmitted due to tumor regrowth. We performed endoscopic biopsy, and histopathologic diagnosis was germinoma. The patient underwent three courses of combined chemotherapy and radiotherapy, and complete response was achieved. Although the precise cause of the transient regression is unknown, racranial germinoma may occasionally unde rgo spontane ous regression. PMID- 19306649 TI - [Bilateral thalamic glioma in an adult: a case report and review of the literature]. AB - We report a case of a 36-year-old woman who had a rare bilateral thalamic glioma (BTG). She complained of memory disorder T1-weighted magnetic resonance imaging revealed enlarged bilateral thalami with homogenous isointensity and no contrast enhancement. Histological examination of the biopsy specimen identified diffuse astrocytoma (WHO grade II). BTG is a rare variant of thalamic neoplasms, which can be distinguished clinically and radiologically from other gliomas. In most of the reported cases, the presenting symptoms were cognitive impairment varying from personality changes to frank dementia. Death usually occurs within two years after onset, independently of adjuvant therapy such as radiotherapy and chemotherapy. On neuroimaging, all of the BTG had a similar appearance, with both thalami being symmetrically enlarged. Our patient has been given radiotherapy and concominant and adjuvant temozolomide in Stupp's regimen. At the time of this writing (5 months after the consultation), there are no neurological symptoms, and no changes on neuroimaging. PMID- 19306650 TI - [Surgical treatment of non-lesional supplementary motor area epilepsy: two case reports]. AB - A 37-year-old male and a 12-year-old female presented with non-lesional epilepsy arising from the supplementary motor area in the medial frontal cortex, manifesting as daily brief tonic seizures involving the bilateral extremities, which were intractable to multiple antiepileptic medications. During the seizure, there was no impairment of awareness, and the tonus was always higher in the limbs contralateral to the epileptogenic hemisphere in both patients. Scalp EEG detected midline epileptiform discharges. Three-tesla MR imaging using the FLAIR and short tau inversion recovery sequences, interictal IMP-SPECT, IMZ-SPECT, and FDG-PET detected no focal abnormalities in either patient. Invasive intracranial EEG evaluation was performed using chronically implanted subdural electrodes on the interhemispheric frontoparietal cortices, based mainly on the seizure semiology and the associated EEG findings. Intracranial EEG successfully identified the location of the seizure onset in the supplementary motor area in the medial frontal cortex. Focal cortical resection including the area of seizure onset resulted in freedom from seizures and significant improvement in both patients. Histological study revealed focal cortical dysplasia in both patients. Recognition of the typical seizure semiology is very important in the correct diagnosis and surgical treatment of supplementary motor area epilepsy, especially in the absence of MR imaging lesions. PMID- 19306651 TI - [Clinical standard of neurosurgical disorder (4). Peripheral and central diseases presenting isolated dizziness or vertigo]. PMID- 19306652 TI - [Molecular mechanism of rhythmic clock gene transcription, lipid metabolism and torpor]. PMID- 19306653 TI - [Roles of the prostanoids in the cardiovascular diseases]. PMID- 19306654 TI - [Alternative menaquinone biosynthetic pathway operating in microorganisms]. PMID- 19306655 TI - [Basic science and clinical applications of mesenchymal stem cells]. PMID- 19306656 TI - [Inflammation-related oxidized lipid mediators]. PMID- 19306657 TI - [Cellular RNA delivery using carrier peptides]. PMID- 19306658 TI - [Collagen I-induced epithelial mesenchymal transition]. PMID- 19306659 TI - [Requirement of in vivo promoter analyses revealed by the research on endothelial cell-specific gene expression]. PMID- 19306660 TI - Lymphology, medical ignorance/ignoramics and the Nobel connection: Howard Florey, Joshua Lederberg, and Francoise Barre-Sinoussi. AB - Intriguing interludes with Nobel laureates have marked the history of Lymphology. Bounded (or rather unbounded) by a mutual fascination with ignorance and the unknown, their curiosity converged around microbes, infections, and host responses mediated by the lymphatic system. These studies transcended a simple "molecular model of life". PMID- 19306661 TI - Effect of particle size on the lymphatic distribution of 111Indium aminopolystyrene through intrapleural administration. AB - The study examined the impact of size on lymphatic particle distribution through intrapleural (ipl.) administration. Aminopolystyrene of three sizes, 0.29 microm, 2.18 microm, and 11.2 microm were radiolabeled with 111Indium and their biodistributions were evaluated in rats after ipl administration. Animals received either particles of three different sizes (4 mg, 200 microCi/animal) or unconjugated 111Indium as control. The percentage of injected dose (%ID) per organ or sample was determined for left (L) and right (R) mediastinal lymph nodes (LN), blood, lung, and pleural wash. The biodistribution of 2.18 microm 111In aminopolystyrene was further investigated at 6 h, 24 h, 48 h, and 72 h following ipl administration to examine the possible particle retention time. The 2.18 microm particles had significantly higher uptake in both LLN and RLN compared to other sizes. The systemic uptake was minimal. At 72 h, there was still 3.2 +/- 3.2% and 2.1 +/- 1.8% of injected dose retained in the LLN and RLN, respectively. Scintigraphic imaging revealed significant accumulation of the radioactivity in mediastinal nodes. Particle size has significant impact on lymphatic particle distribution through ipl administration. Approximately 2 microm seems to be a suitable size. PMID- 19306662 TI - Complex decongestive physiotherapy decreases capillary fragility in lipedema. AB - Lipedema is a disproportional obesity featuring frequent hematoma formation due to even minor traumatic injuries. On the basis of clinical observations, complete decongestive physiotherapy diminishes the incidence of hematomas due to minor injuries beyond leg volume reduction. Hematoma development may be caused by altered capillary resistance (CR) or altered capillary fragility (CF). We measured capillary fragility (CF) before and after complex decongestive physiotherapy (CDP) to examine, whether CDP could reduce CF. 38 women with lipedema were included in the study. Twenty-one (21) patients were treated with CDP and 17 using exclusively moisturizers as the control group. CDP comprised once daily manual lymph drainage, intermittent pneumatic compression and multilayered short-stretch bandaging performed throughout a 5-day-course. CF was evaluated with the vacuum suction method (VSM) using Parrot's angiosterrometer in both groups. Decongestive therapy resulted in a significant reduction of the number of petechiae while no change was detected within the control group. Complete decongestive physiotherapy significantly reduced CF in patients with lipedema and this reduction may lead to reduced hematoma formation. PMID- 19306663 TI - Measurement of soft tissue compliance with pressure using ultrasonography. AB - Lymphedema is swelling, particularly in the subcutaneous tissues, due to accumulation of lymph. Previous imaging techniques have demonstrated associated structural changes and have been used for evaluating the status of soft tissues. However, the reliability of measurements using ultrasonography has not been evaluated and the ultrasonographic method has been unable to show changes in tissue softness. There is a need to determine if ultrasonography is a reliable technique to assess skin and subcutaneous thickness in the upper extremities with or without pressure and if the measure of compliance in soft tissue thickness is also reliable. Two examiners measured tissue thicknesses using ultrasonography and calculated the compliance with and without pressure on the forearm and upper arm, independently. The intra- and inter-rater cross-correlation coefficients of measuring soft tissue thickness were excellent (> 0.75) in the forearm (p < 0.05). In the upper arm, the reliabilities were fair-to-good. The intraclass correlation coefficients of pressure compliance in the forearm and upper arm were excellent and fair-to-good, respectively (p < 0.05). This study suggests that measurement of thickness of soft tissues using ultrasonography may be reliable and pressure compliance may reflect tissue softness in the upper extremity. PMID- 19306664 TI - Contrast-enhancing optical method to observe a Bonghan duct floating inside a lymph vessel of a rabbit. AB - Novel threadlike structures, so-called Bonghan ducts (BHDs), were recently rediscovered inside large caliber lymphatic vessels using two different staining dyes in rabbits (Janus green B and Alcian blue) and fluorescent nanoparticles in rats. These three methods have the drawback of injecting chemical agents into the lymphatic vessels, which might damage the BHD and hinder further investigation of its physiological function. New methods to observe BHDs without using external chemical agents need to be developed. In the present work, we introduce a contrast enhancing optical method for in vivo observation of BHDs floating inside large caliber lymph vessels. The method uses a low-pass filter above about 650 nm, with an arrangement to minimize the light reflected from the surface of the lymph vessel. We captured films showing movement of a BHD as the animal respired. Applying the previous Alcian blue injection technique, we obtained BHD samples from the lymph vessel and observed the distribution of rod-shape nuclei (the essential feature of a BHD). BHDs can now be observed inside lymph vessels by using contrast-enhancing instrumentation without visualizing chemical agents. PMID- 19306665 TI - Local tissue water changes assessed by tissue dielectric constant: single measurements versus averaging of multiple measurements. AB - Previous reports describe the use of average tissue dielectric constant (TDC) measurements to assess local tissue water and its change. Our goal was to determine if a single TDC measurement could be used in place of the average of multiple measurements. The comparison criteria used to test this was the extent to which single and averaged measurements yielded similar TDC values in both normal and lymphedematous tissue. Measurements were made on both ventral forearms to a depth of 2.5 mm in 10 women with unilateral arm lymphedema. The main results showed that the 95% confidence interval for differences between single and averaged TDC values was less than +/- 1 TDC unit for both normal and lymphedematous arms. This finding strongly suggests that for most, if not all, clinical evaluations, suitable assessments can be made using a single TDC measurement. PMID- 19306666 TI - Naming names for unified agency. PMID- 19306667 TI - MRSA: a crisis that touches you. How automated molecular dx will help relieve the time and cost crises facing hospital labs. PMID- 19306668 TI - Labs take new role in antibiotic stewardship, championing improved diagnoses, guiding therapy, saving lives, and cutting costs. PMID- 19306669 TI - Implementing a real-time PCR assay for rapid surveillance of MRSA. PMID- 19306670 TI - Identity in the PALM of your hand. PMID- 19306671 TI - The staffing agency "scoop" on how to find employment. PMID- 19306672 TI - An educator's "scoop" on asking for or writing a recommendation. PMID- 19306673 TI - Addressing management issues: not following protocol. PMID- 19306674 TI - Reporting impaired pathologist. PMID- 19306675 TI - "Molecular" is not your traditional lab: where disease management meets acute care. PMID- 19306676 TI - Ways of knowing and willingness to argue. AB - Willingness to argue is associated with higher level thinking. The authors tested the relation between ways of knowing-involving separate knowing (i.e., playing the devil's advocate) and connected knowing (i.e., empathic understanding)--and students' willingness to argue. Participants were 171 male and 231 female college undergraduates who completed assessments in ways of knowing and willingness to argue. Also, the participants defined the word argument in their own words. After the authors controlled for demographic variables, endorsement of separate knowing predicted willingness to argue. Students with high scores in separate knowing (objective, adversarial knowing) and connected knowing (subjective, empathic knowing) indicated more willingness to argue. Furthermore, these same students defined argument as a constructive form of communication. Students with low scores in separate knowing defined argument as an emotional battle with the goal of psychological harm. This negative perspective could be an impediment to engaging students in classroom debate and critical thinking. PMID- 19306677 TI - Assessing organizational citizenship behavior in the French context: evidence for the four-dimensional model. AB - Although researchers have extensively studied organizational citizenship behavior (OCB) in the U.S. context, OCB measurement has received relatively limited attention in other contexts, specifically in the French-language context. Using 2 samples (for Study 1, N=292; for Study 2, N=355), the author investigated OCB in a French-language context. Using an exploratory factorial analysis in Study 1, the author found a 4-factor model (altruism, civic virtue, sportsmanship, helping others). Using a confirmatory factor analysis in Study 2, the author confirmed a 4-factor model. Data provided results that show some differences and similarities between U.S. and French contexts. PMID- 19306678 TI - Psychological aspects in continuous subcutaneous insulin infusion: a retrospective study. AB - The authors aimed to analyze the relation of psychological predictors with medical and psychological therapy success indicators in continuous subcutaneous insulin infusion (CSII). Besides blood glucose control as a medical indicator of therapy success (by means of HbA(1c) levels), the authors assessed treatment satisfaction, depressive symptoms, and quality of life among 51 adult patients on CSII. The authors examined the following psychological factors that were assumed to be associated with outcome variables: coping style, locus of control, and self efficacy. Self-efficacy correlated positively with all psychological outcome variables. A high external locus of control was related to negative glycaemic outcome and more depressive symptoms. Multiple linear regression analyses revealed that when considering all psychological factors simultaneously, locus of control was the strongest predictor of HbA(1c). For the psychological outcomes, self-efficacy was the most powerful predictor. The authors demonstrated that medical and psychological diabetes-related outcomes were strongly related to psychological factors in adult CSII patients. Patients who felt responsible for the process of CSII therapy showed more positive outcomes; therefore, psychological care can be seen as an important part of diabetes education in CSII therapy. PMID- 19306679 TI - Can mock interviewers' personalities influence their personality ratings of applicants? AB - The authors examined individual difference and self-regulatory variables to understand how an interviewer rates a candidate's personality. Participants were undergraduate students at a large midwestern university in the United States who completed measures of individual differences, read an employment interview transcript involving a candidate applying for a customer service job, and rated the candidate's personality. Participants' agreeableness, social skills, and communion striving were positively associated with their ratings of the candidate's helpfulness and obedience. The authors provide a foundation for further research on interviewer effectiveness and the processes underlying the employment interview. PMID- 19306680 TI - Assessing competition avoidance as a basic personality dimension. AB - The lack of an adequate psychometric instrument has impeded personality theory testing in the area of competition avoidance. The authors conducted 6 studies in an attempt to remedy this deficiency by constructing an individual-difference measure of competition avoidance. In line with K. Horney's (1937) interpersonal theory of neurosis, participants who were higher in competition avoidance showed higher levels of neuroticism, greater fears of both success and failure, a lower desire to prove themselves in competitive situations, higher levels of self handicapping, and more maladaptiveness than those participants who were lower in competition avoidance. Also, they were more modest and willing to conform to group standards than were those lower in competition avoidance. The authors discuss the measure's potential usefulness as a diagnostic and assessment tool in academic, athletic, and clinical settings. PMID- 19306681 TI - Activities of daily living, depression, and social support among elderly Turkish people. AB - The authors examined the effects of activities of daily living (ADL) and perceived social support on the level of depression among elderly Turkish people. Participants were 102 adults older than the age of 60 years. The authors hypothesized that (a) lower levels of ADL would predict a higher level of depression, (b) a higher level of perceived social support would predict a lower level of depression, and (c) perceived social support would moderate the relation between ADL and depression. Although hierarchical multiple regression analysis did not yield a significant effect for an ADL-perceived social support interaction, ADL and perceived social support significantly predicted depression among elderly people. Thus, perceived social support did not moderate the relation between ADL and depression among elderly people; however, higher ADL functioning and higher perceived social support predicted lower depression. The nonsignificant effect of an ADL-perceived social support interaction on the level of depression among elderly people was incongruent with the stress-buffering model (S. Cohen & T. A. Willis, 1985). The authors discuss the strengths, limitations, and possible implications of the findings. PMID- 19306682 TI - Implicit letter preferences in job choice: an experimental test of the role of cognitive load. AB - Research has shown that people prefer the letters in their names to letters that are not in their names. This name-letter effect seems to influence important life decision such as where one chooses to live or whom one chooses to marry. The authors' laboratory study investigated whether this effect generalizes to individuals' job-choice intentions under specific conditions. Furthermore, the authors hypothesized that name-letter preferences in job-choice intentions would be stronger under conditions of high cognitive load than under conditions of low cognitive load. Two experiments with final-year students attending a university in Belgium showed support for name-letter preferences in job-choice intentions. There was no support for the hypothesized moderating role of cognitive load. The authors discuss the implications of these results for theory and research on name letter preferences and job choice. PMID- 19306683 TI - [Localization and dynamic change of saponin in root tuber of cultivated Pseudostellaria heterophylla]. AB - Anatomical, histochemical and phytochemical methods were used to investigate the structure, the localization and content changes of total saponin of the root tuber of Pseudostellaria heterophylla on different developmental stages. Results showed that the primary structure and secondary structure of P. heterophylla adventitious root were similar to that of other herbaceous dicotyledons. 80% of mature root tuber were secondary xylem which consisted of most parenchyma cells and less vessels. The secondary phloem were composed of parenchyma cells too. The results of histochemical methods showed that saponin distributed in pericycle and parenchyma cells of primary phloem in the primary structure of root. In secondary structure and mature root tuber, saponin distributed in periderm and secondary vascular structure except cork layer and vessel. Besides, the colors of the secondary phloem were darker. The results of phytochemical test showed that the content of saponin in skin areas was higher than that in xylems in February and July. The result was consistent with that of histochemistry. The content of saponin in the head of the root was higher than that in the end of the root which was in turn higher than that in the middle part of the root. In the developmental process of root tuber, the content of saponin showed a dynamic tendency of high low-high. This characteristics were related to the development of root system. PMID- 19306684 TI - [Molecular cytogenetic identification of Aegilops ventricosa x Aegilops cylindrica amphiploid SDAU18]. AB - SDAU18, an amphiploid of Ae.ventricosa with Ae.cylindrica, was identified by cytological analysis, seed storage protein electrophoresis, genomic in situ hybridization (GISH) and inoculation assessment. The results are as follows: The chromosome number of root tip cells (RTCs) of SDAU18 plants varied from 52 to 56. 28 bivalents were observed in most PMCs MI of SDAU18 with 56 chromosomes, meanwhile, a few univalents, multivalents also existed in some PMCs MI, and the average chromosome configuration was 2n = 56 = 3.21 I +19.78 II, (Ring)+6.50 II (Rod)+0.01 III +0.04 IV (Ring)R+0.01 IV (Rod). There were both Ae. ventricosa specific bands and Ae. cylindrica-specific bands in the seed storage protein electrophoretogram of SDAU18, furthermore, SDAU18 had one novel HMW-GS not found in the parents and two novel ones not found in common wheats. By labeling the total genomic DNA of Ae. ventricosa and Ae. cylindrica as probes respectively, and using that of another parent as block, GISH of RTCs spread of SDAU18 was carried out. The green hybridization signal was observed in 14 chromosomes respectively, within 56 ones in RTCs of SDAU18. SDAU18 was immune to powdery mildew and stripe rusts. SDAU18 was an amphiploid of Ae. ventricosa with Ae. cylindrica, and had very important significance in wheat breeding and genetic improvement. PMID- 19306685 TI - [The change of learning and memory ability in the rat model of depression]. AB - The present study was performed to explore learning and memory ability in the depression models of rats. The chronic unpredictable stress and olfactory bulbectomy model of rats were adopted. Open-field test was used to detect the locomotion activity and HPLC-UV was employed to analyze the level of blood serum cortisol. The method of Morris water Maze was used to measure learning and memory ability and the results of LTP and LTD in hippocampus CA1 were recorded to observe the synaptic plasticity of hippocampus neurons. The results showed that compared with control group, the locomotion activity and learning ability for two models decreased extremely, while there was no apparent difference in the feedback of memory. Meanwhile, the synaptic plasticity of hippocampus neurons for two models decreased significantly and the level of serum cortisol increased evidently. These results suggested that both methods employed to build the models could cause rats depression and learning inhibition, but do no effects on the feedback of memory. PMID- 19306686 TI - Auxin-regulated OsRGP1 and OsSuS are involved in gravitropic bending of rice shoot bases. AB - Gravitropic bending of horizontally-oriented rice shoots results from the differential elongation of cells in the upper and lower halves of the shoot bases. In this study, genes encoding a reversibly glycosylated polypeptide (OsRGP1) and a sucrose synthase (OsSuS) related to sugar metabolism were identified by suppressive subtractive hybridization (SSH) as being differentially expressed in gravibending lower and upper halves of rice shoot bases. RT-PCR was used to monitor OsRGP1 and OsSuS gene expression. The two genes were differentially induced in lower and upper halves of the shoot bases during gravitropism and their expression was regulated by auxin. Gene promoter sequence analysis demonstrated the existence of elements related to auxin. Treatment with the auxin transport inhibitor TIBA inhibited the asymmetric expression of OsRGP1 and OsSuS. In addition, an increase in hexose sugars was detected in the lower half of the shoot bases during gravitropism. Our data suggest that asymmetric redistribution of auxin following gravistimulation results in differentially localized OsRGP1 and OsSuS expression. While asymmetric expression of OsSuS may result in a disproportionate distribution of hexose, asymmetric expression of OsRGP1 could induce cell wall polysaccharide synthesis in the lower half of shoot bases. Therefore hexose and cell wall polysaccharide accumulation in the lower half of rice shoot bases might contribute to cell expansion and subsequent gravitropic bending. PMID- 19306687 TI - [EoRab43 regulating vesicular transport around the macronucleus in Euplotes octocarinatus]. AB - Rab family proteins play a crucial role in regulating vesicular traffic in eukaryotic cells. EoRab43 is an atypical Rab gene identified in Euplotes octocarinatus. In order to understand the function of EoRab43, the 153bp fragment of the 3'-end of EoRab43 gene was subcloned into expression vector pGEX-6P-1, and the recombinant plasmid pGEX-EoRab43(153bp) was transfered into E.coli BL21 (DE3) to express the fusion protein. The fusion protein GST-EoRab43C was expressed and purified by affinity chromatography. BALB/c mice were immunolized by purified GST EoRab43C. The titer of anti-EoRab43C polyclonal antibody was detected by indirect ELISA assay and the specificity of the antibody was detected by Western blot. Immunofluorescence experiments were performed using anti-EoRab43C antibody in the cells of Euplotes. The results showed that EoRab43 displayed a punctuate pattern in the cytoplasm around the macronuclear chromosome of Euplotes. PMID- 19306688 TI - Expression inhibition of multidrug resistance-associated protein (MRP1) by multi ribozyme expression system in HEK293 cells. AB - To study application of multi-ribozyme expression system on expression inhibition of multidrug resistance-associated protein (MRP1) in HEK293 cells, the multi ribozyme expression system containing 20 cis-acting ribozymes for self-cleavage and 10 trans-acting ribozymes for targeting to MRP1 gene specific site were constructed. HEK293 cells cotransfected multi-ribozyme expression system with MRP1 target gene or full length of MRP1 gene respectively were analyzed by RT PCR, Western blot analysis and MTT assay. The results showed that multi-ribozyme systems were able to dramatically decrease fluorescent fusion protein expression in HEK293 cells. RT-PCR analysis indicated that the extent of MRP1 target mRNA decrease was correlated with the number of trans-acting ribozyme contained in multi-ribozyme expression system. Similar changes have been observed from Western blot. MTT assay showed that multi-ribozyme systems were able to reverse MDR generated by MRP1 gene in HEK cells. These results suggested that inhibitory effects of multiple copies of ribozymes contained in the system were better than that of single ribozyme contained. Therefore, this strategy could be used in treatment of tumor or other diseases via gene therapy. PMID- 19306689 TI - [A genome-wide analysis of P450 gene family in waterflea (Daphnia pulex)]. AB - A genome-wide analysis of P450 genes was performed in waterflea genome based on their mRNA and amino acid data. Seventy-one P450 genes, assigned into 15 gene families and 17 subfamilies, were identified. They were partitioned into 12 groups and 16 orphan genes by a criterion that the genes belong to the same group when their amino acid identity was more than 60%. Except orphan genes and 4 gene groups, including 2 genes per group, positive selection and gene conversion were analyzed in other 8 groups, including 47 genes. The results showed that positive selection was detected in 3 groups (including 22 genes). The positively selected sites 228E or 277T, locating in SRS (substrate recognition sites) 2 or SRS 5, were detected in 2 groups (18 sequences) with a more than 95% probability, and gene conversions were detected in 5 groups (19 sequences). Interestingly, both positive selection and gene conversion events were identified obviously in 12 genes, and neither positive selection nor gene conversion event was detected in 18 genes. The correlation was revealed between positive selection and gene conversion: positive selection sites were identified in 63.2% of genes in which gene conversion events were detected, and gene conversion events were identified in 54.5% of genes in which positive selections were detected. Moreover, twenty motifs were identified, there in to, 5 motifs existed in over 90% of P450 genes. PMID- 19306690 TI - [The structure of vegetative organs, and saponins histochemical localization and content comparization in Polygala sibirica L]. AB - Anatomical, histochemical and phytochemistry methods were used to investigate the structure of vegetative organs, and saponins localization and dynamic changes in Polygala sibirica L. The root consisted of developed periderm and secondary vascular. The secondary phloem was thick, and mainly composed of parenchyma. There were well-developed vessels and fibers in the secondary xylem. The stem was composed of epidermis, cortex and vascular bundle. The ring of sclerenchymatous cells lied between cortex and phloem might be the apoplastic protective screen which could protect the stem from drought. The leaf was bifacial one. The root and stem possessed characteristics adapting to arid environment. Histochemical localization results showed that saponins distributed in secondary phloem and phelloderm of root, in epidermis, cortex and phloem of stem, mainly in mesophyll of leaf. It displayed that saponins accumulated mainly in parenchyma cells of vegetative organs, among of which, the secondary phloem was the main storage site. The HPLC results also showed that the saponins accumulated in all the vegetative organs of Polygala sibirica L., with higher content in roots and lower content in the aerial part that included stems and leaves. The study indicated the aerial part of Polygala sibirica L. also had medicinal value. The saponins content had dynamic variance at the developmental stage, the crude drug should be gathered at period from April to May. PMID- 19306691 TI - [Establishment of loop-mediated isothermal amplification for detection of Vibrio alginolyticus]. AB - Vibrio alginolyticus is the main Vibrio pathogen in aquaculture in the south of China. A one step loop-mediated isothermal amplification (LAMP) assay was developed for detection of Vibrio alginolyticus. A set of primers were designed from the OmpK sequence of Vibrio alginolyticus. The assay was optimised to amplify Vibrio alginolyticus DNA by incubation at 65 degrees C for only 1 h, and required only a simple water bath or heating block to provide a constant temperature of 65 degrees C. LAMP amplification products had a ladder-like appearance when electrophoresed on an agarose gel. The detection limit of the LAMP assay was n(cell) = 38/mL which was found to be higher than the commonly used PCR method. The assay was evaluated using clinical samples and the results indicated the suitability and simplicity of the test as a rapid, field diagnostic tool for Vibrio alginolyticus. PMID- 19306692 TI - [Analysis of differential protein expression of REMI mutant of Trichoderma koningii]. AB - On purpose of expounding the phenotype change of Trichoderma mutant which produces cellulase, protein profiles of T. koningii CICC and its mutant TK-2R-1 were detected by 2-DE. Eight unique spots were found and four of them were identified and their functions by MS-TOF-TOF were connected with growth and metabolization of Trichoderma spp. The research provides a new theoretical basis and methods for the construction of Trichoderma mutant which is used to produce efficient cellulase. PMID- 19306693 TI - [Hydrogen peroxide is involved in regulation of tobacco leaf senescence]. AB - Effects of various exogenous hydrogen peroxide (H2O2) concentrations on leaf senescence of tobacco (Nicotiana tabacum L.) were studied. The results showed that 0.1 and 1 mmol/L H2O2 had little effects on the leaf senescence of tobacco, the contents of chlorophyll and soluble protein were decreased in varying degrees by 10, 100 mmol/L H2O2 treatment, the decreased chlorophyll and soluble protein contents were significantly correlated with increased endogenous H2O2 content. SDS-PAGE of soluble protein and localized staining of H2O2 showed consistency with respective content. Chloroplast ultrastructure were observed in the mesophyll cells of tobacco and there was a little change by 10 mmol/L H2O2 treatment, and a remarkable breakdown of granal stacks and disorganized thylakoid membranes were revealed by 100, 200 mmol/L H2O2 treatment. RT-PCR analysis showed the expression level of senescence associated genes SAG12 was low by 10mmol/L H2O2 treatment and increased by 100 mmol/L H2O2 treatment. These results strongly suggested the tobacco leaf senescence was induced by 10 mmol/L H2O2 and accelerated the progression by high concentration (100, 200 mmol/L) H2O2 stress. PMID- 19306694 TI - Moral complicity in induced pluripotent stem cell research. AB - Direct reprogramming of human skin cells makes available a source of pluripotent stem cells without the perceived evil of embryo destruction, but the advent of such a powerful biotechnology entangles stem cell research in other forms of moral complicity. Induced pluripotent stem cell (iPSC) research had its origins in human embryonic stem cell research and the projected biomedical applications of iPS cells almost certainly will require more embryonic stem cell research. Policies that inhibit iPSC research in order to avoid moral complicity are themselves complicit in preventable harms to patients. Moral complicity may be unavoidable, but a Blue Ribbon Panel charged with assessing the need for additional embryonic stem cell lines may ease a transition from embryonic stem cell research to clinical applications of iPS cells. PMID- 19306695 TI - Ontological and ethical implications of direct nuclear reprogramming. AB - Scientific breakthroughs rarely yield the potential to engage a foundational ethical question. Recent studies on direct reprogramming of human skin cells reported by the Yamanaka lab in Japan and the Thomson lab in Wisconsin suggest that scientists may have crossed both a scientific and an ethical threshold. The fascinating science of direct nuclear reprogramming highlights empirical data that may clarify the ontological status of cellular activity in the early stages of what could become a human fetus and justify ethical options for research in this controversial field. The ontological and ethical implications that accrue here are connected with the biological or natural potentiality of these cells. PMID- 19306696 TI - The HFEA public consultation process on hybrids and chimeras: informed, effective, and meaningful? AB - In September 2007, the Human Fertilisation and Embryology Authority (HFEA) in the United Kingdom concluded that "there is no fundamental reason to prevent cytoplasmic hybrid research ... this area of research can, with caution and careful scrutiny, be permitted." Later, in January 2008, HFEA issued two research licenses to create humanesque cytoplasmic hybrid embryos from which stem cells could be derived. This article critically examines the public consultation process that preceded these decisions, concluding that the process was flawed and demonstrating how the HFEA documents summarizing the findings of the public consultation process misrepresent the public's contributions to this policymaking initiative. PMID- 19306697 TI - Deliberative Democracy and stem cell research in New York State: the good, the bad, and the ugly. AB - Many states in the U.S. have adopted policies regarding human embryonic stem cell (hESC) research in the last few years. Some have arrived at these policies through legislative debate, some by referendum, and some by executive order. New York has chosen a unique structure for addressing policy decisions regarding this morally controversial issue by creating the Empire State Stem Cell Board with two Committees--an Ethics Committee and a Funding Committee. This essay explores the pros and cons of various policy arrangements for making public policy decisions about morally controversial issues in bioethics (as well as other issues) through the lens of Deliberative Democracy, focusing on the principles of reciprocity, publicity, and accountability. Although New York's unique mechanism potentially offers an opportunity to make policy decisions regarding a morally controversial subject like hESC research in accord with the principles of Deliberative Democracy, this essay demonstrates its failure to do so in actual fact. A few relatively simple changes could make New York's program a real model for putting Deliberative Democracy into practice in making policy decisions regarding controversial bioethical issues. PMID- 19306698 TI - Future directions for oversight of stem cell research in the United States. PMID- 19306713 TI - Adult herpes simplex encephalitis: fifteen years' experience. AB - INTRODUCTION: Herpes simplex encephalitis (HSE) is the most frequent cause of sporadic necrotizing encephalitis in adults. The aim of this study is to describe the characteristics of HSE and the factors influencing its outcome. MATERIAL AND METHODS: Retrospective study of patients diagnosed with HSE in a tertiary care teaching hospital over a 15-year period. Diagnosis was based on a consistent clinical profile for HSE, plus either a PCR-positive CSF HSV study or consistent brain neuroimaging findings. Patients were divided into 2 groups according to the modified Rankin Scale: good outcome (Grades <=2) and poor outcome (Grades >=3). RESULTS: Thirty-five patients were included. Mean age was 53.9 years. More than half presented febricula or fever, headache, disorientation, behavioral changes, decreased level of consciousness, or neurological deficit. CSF glucose concentration was normal in all patients and WBC count was normal in 8 (23%). PCR for HSV was positive in 92% and cranial MRI was suggestive of HSE in 100% of patients. Mortality was 8.6%. In relation to outcome, age (OR=1.079; 95% CI, 1.023-1.138) and serum albumin level at admission (OR=0.87; 95% CI, 0.794-0.954) were independent prognostic factors at discharge. At 6 months, days of fever after initiation of acyclovir therapy (OR=1.219; 95% CI, 1.046-1.422) and serum albumin level at admission (OR=0.917; 95% CI, 0.87-0.967) were independent prognostic factors. CONCLUSIONS: Normal brain MRI or detection of low CSF glucose concentration requires consideration of diagnoses other than HSE. Age, serum albumin level at admission, and days of fever after initiation of acyclovir therapy were independent prognostic factors of the disease. PMID- 19306714 TI - [Evaluation of methods for studying susceptibility to oxacillin and penicillin in 60 Staphylococcus lugdunensis isolates]. AB - INTRODUCTION: Staphylococcus lugdunensis is a coagulase-negative staphylococcus associated with a variety of clinical infections. In this paper we present the results of a comparative study using 4 methods to determine antimicrobial susceptibility to oxacillin and penicillin in 60 S. lugdunensis isolates. MATERIAL AND METHODS: We studied 60 S. lugdunensis isolates obtained from clinical specimens sent to our laboratory over an 8-year period. All isolates were free coagulase-negative and DNase-negative, and biochemically identified by API ID 32 STAPH (bioMerieux). Presence of mecA and ss-lactamase production were studied in all cases. Antimicrobial susceptibility was determined by the Vitek 2 System (bioMerieux) and broth microdilution (Wider) (Soria Melguizo) for penicillin and oxacillin, and the E-test (AB Biodisk) and cefoxitin disk diffusion test (BD BBLTM) for oxacillin. RESULTS: All isolates lacked the mecA gene and were susceptible to oxacillin by broth microdilution, E-test, and cefoxitin disk diffusion test. Only two isolates were oxacillin-resistant by the Vitek 2 System. Twenty-four isolates (40%) were ss-lactamase-positive, 4 after induction. Susceptibility testing to penicillin determined that 48 isolates showed concordance between the results obtained by broth microdilution and Vitek 2, but 12 isolates (20%), showed divergent results. CONCLUSIONS: We detected no resistance to oxacillin in S. lugdunensis. All the methods evaluated were adequate for determining oxacillin resistance. The Vitek 2 System is useful for detecting penicillin resistance, but the ss-lactamase test should be applied to isolates with a MIC=0.25microg/ml to avoid the interpretation of false resistance to this antibiotic. PMID- 19306715 TI - [Correlation between antibiotic prescription and decreased antimicrobial resistance in the health area of Zamora]. AB - OBJECTIVE: To describe the trends in prescribing antibiotics and analyze a possible relationship with the evolution of antimicrobial sensitivity in respiratory pathogens isolated in the health area of Zamora (Spain). MATERIALS AND METHODS: Descriptive study investigating the prescription of antimicrobials and the evolution of resistance in Streptococcus pyogenes, Streptococcus pneumoniae and Haemophilus influenzae, from 1996 to 2005. Antibiotic use is expressed in doses per 1,000 inhabitants per day (DHD). The chi(2) test, exact tests, Spearman correlation coefficients, and a time series analysis were used to analyze the trends. RESULTS: A total of 1301 S. pyogenes strains were isolated. All were sensitive to penicillin, with decreasing percentages of resistance to erythromycin and increasing percentages of resistance to clindamycin. Among 579 pneumococcal strains isolated, a significant decreasing resistance to penicillin, amoxicillin-clavulanic, cefotaxime and cefuroxime was observed; resistance to erythromycin and azithromycin increased during the first few years, but decreased later. Among a total of 769 strains of H. influenzae, all were sensitive to cefotaxime and highly sensitive to amoxicillin-clavulanic acid and cefuroxime; resistance to ampicillin was maintained and there was a significant decreasing resistance to azithromycin. Simultaneously, there was a progressive decrease in overall antibiotic use (20.45 DHD in 1996; 18.90 DHD in 2005). Prescription of penicillins remained stable, with increased use of amoxicillin-clavulanic, decreased use of cephalosporins and macrolides (except azithromycin), and increased use of fluorquinolones. We observed a significant correlation between prescription and resistance. CONCLUSIONS: A recovery of sensitivity to antibiotics was seen in respiratory pathogens parallel to a progressive decrease in prescription of antibiotics. PMID- 19306716 TI - [Factors associated with prolonged hospital stay in community-acquired pneumonia]. AB - INTRODUCTION: The length of hospital stay in patients with community-acquired pneumonia (CAP) varies considerably, even though this factor has a great impact on the cost of care for this condition. The objective of this study was to identify factors associated with prolonged hospitalization in these patients (>8 days). METHODS: Observational analysis of a prospective cohort of nonimmunosuppressed adults with CAP requiring hospitalization from 1995 through 2006. RESULTS: We documented a total of 2688 consecutive episodes of CAP. Patients who required intensive care unit admission from the emergency room (n=107), those who died during hospitalization (n=200), and patients hospitalized for more than 30 days (n=60) were excluded from the analysis. The median duration of hospital stay was 8 days (IQR, 6-11). Factors independently associated with prolonged hospital stay by stepwise multiple logistic regression analysis were advanced age (OR=1.58; 95% CI, 1.002-2.503), alcohol abuse (OR=2.07; 95% CI, 1.341-3.199), high-risk Pneumonia Severity Index class (OR=1.72; 95% CI, 1.094 2.703), aspiration pneumonia (OR=4.57; 95% CI, 1.085-19.285), pleural empyema (OR=3.73; 95% CI, 1.978-7.04), and time to clinical stability (OR=1.13; 95% CI, 1.065-1.196). CONCLUSIONS: Several factors that were independently associated with longer hospital stay in adult patients with CAP. These factors should be considered when evaluating the adequacy of the duration of hospitalization in a specific center and when designing future studies investigating new strategies to reduce the length of hospital stay. PMID- 19306717 TI - [Prenatal screening for Trypanosoma cruzi and human T lymphotropic virus types 1 and 2 in pregnant Latin American women]. AB - INTRODUCTION: To estimate the seroprevalence of infection by Trypanosoma cruzi and human T lymphotropic virus (HTLV) in pregnant Latin American women. METHODS: Serological survey carried out in pregnant Latin American women attending the antenatal care clinic of a Spanish hospital from January 2006 to June 2007. RESULTS: Of the 229 women enrolled, 4 had antibodies against T. cruzi (1.75%; 95% confidence interval [95% CI], 0.68-4.4); 2 of these women came from Bolivia (13.33%; 95% CI, 3.73-37.88) and the other 2 from Paraguay (11.76%; 95% CI, 3.29 34.33). None of the women had anti-HTLV-1 antibodies (95% CI, 0-1.6), and 2 had HTLV-2 antibodies (0.87; 95% CI, 0.24-3.12). CONCLUSIONS: A notable percentage of pregnant immigrant women from Latin American had T. cruzi infection. The seroprevalence of HTLV infection is low. PMID- 19306718 TI - [Microbiological diagnosis of viral respiratory infections]. AB - Acute respiratory infection is the most common disease occurring over a person's lifetime, with etiological variations determined mainly by age, environmental circumstances, the healthcare setting, and the underlying pathology. More than 200 different viruses distributed in six viral families have been implicated in the pathogenesis of respiratory tract infection. These facts are generating an increasing diagnostic demand that should be incorporated into the healthcare setting without delay. To meet this demand, the Spanish Society of Infectious Diseases and Clinical Microbiology has updated its Standard Procedure for the microbiological diagnosis of viral respiratory infection. This document contains an update primarily of infections caused by influenza viruses, and secondarily, infections due to other conventional and emerging respiratory viruses. In all cases, the methods for direct virological diagnosis (cell culture, and detection of antigens and nucleic acid) are reviewed, with special reference to techniques for molecular detection and genetic characterization. PMID- 19306719 TI - [Non-specific cutaneous lesion in a 17-year-old pregnant adolescent]. PMID- 19306720 TI - [Streptococcus agalactiae tricuspid valve endocarditis. A case report and review of literature]. PMID- 19306721 TI - [Severe aortic regurgitation secondary to Erysipelothrix rhusiopathiae endocarditis]. PMID- 19306722 TI - [Cerebral aspergillosis in an HIV-infected patient: unsuccessful outcome despite combined antifungal therapy. ]. PMID- 19306723 TI - [Streptococcus suis meningitis in Spain]. PMID- 19306727 TI - Allografts. Preface. PMID- 19306728 TI - Current safety sterilization and tissue banking issues for soft tissue allografts. AB - Allografts used in orthopedic surgery have steadily increased. With concerns regarding tissue safety and processing, governing entities have increased their regulation. This review articles discusses current testing and processing of allografts as well as the rules of their handling. PMID- 19306729 TI - Tissue processing: role of secondary sterilization techniques. AB - Secondary sterilization of musculoskeletal allografts may use chemicals, radiation, or combinations of these. No sterilization techniques have been definitively proven to be more effective than others, and their biomechanical and biological effects on allograft tissue remain largely unknown. The current risk of an allograft infection appears to be much less than the risk of infection surrounding the surgical procedure itself. With appropriate donor screening, improved donor testing--including nucleic acid testing (NAT), and adherence to AATB standards--the risk of disease transmission or infections can be eliminated or substantially decreased. PMID- 19306730 TI - Biology of allograft incorporation. AB - The use of implantation of allograft tissue in sports medicine has risen steadily over the last decade. Allograft tissues offer several advantages over autografts, including reduced donor-site morbidity and decreased operative time, and in some instances, no autograft option exists. However, with allografts, there is a small risk of disease transmission, immunologic rejection, and decreased biologic incorporation. Several techniques to limit these pitfalls and maximize graft incorporation are available, however. This chapter takes an in-depth look at the biology of allograft incorporation and how these techniques affect graft incorporation. PMID- 19306731 TI - What do we really know about allografts? AB - The use of allografts in sports medicine is becoming increasingly popular, and, therefore, this issue of Clinics in Sports Medicine is dedicated to the use of allografts in sports medicine. The majority of indications are related to the use of soft tissue grafts for ligament reconstruction, osteochondral (OC) allografts for articular surface reconstruction, and meniscal allografts for meniscal transplantation. There is an increasing amount of science and literature dealing with healing and outcomes, but many questions still remain. There are a number of issues, controversies, and lack of long-term outcomes to make definitive statements on what is really known about allograft use in sports medicine. PMID- 19306732 TI - Primary ACL reconstruction using allograft tissue. AB - While bone-patellar tendon-bone (BPTB) autograft continues to be the "gold standard" and most popular graft choice for primary anterior cruciate ligament (ACL) reconstructions, the use of allograft tissues in ACL reconstruction has steadily increased over the last 2 decades. Advantages of allograft include a lack of donor-site morbidity, unlimited available sizes, shorter operative times, availability of larger grafts, smaller incisions, improved cosmesis, lower incidence of postoperative arthrofibrosis, faster immediate postoperative recovery, and less postoperative pain. Disadvantages include the potential for disease transmission and prolonged graft healing. Presented in this article are 2 techniques used at the authors' institution for primary ACL reconstruction with allograft. With the proper indications, knowledge of graft preparation and handling, and technique, allograft tissues in ACL reconstructions can provide the surgeon with clinical results equal to those of autograft reconstructions. PMID- 19306733 TI - Posterior cruciate ligament reconstruction: Achilles tendon allograft, double bundle. AB - Allograft usage for cruciate ligament reconstruction has gained in popularity. Many techniques are described for posterior cruciate reconstruction with both autograft and allograft tendons. Achilles tendon allograft is a versatile and effective graft that can be used for a transtibial, double femoral bundle posterior cruciate reconstruction. PMID- 19306734 TI - Meniscal allograft transplantation. AB - Meniscal allograft transplantation has emerged as a treatment option for selected meniscus-deficient patients to restore normal meniscal function and forestall progressive joint degeneration. Contraindications include diffuse subchondral bone exposure, axial malalignment, and instability. However, a knee may be rendered suitable for meniscus transplantation if combined with chondral resurfacing, osteotomy, and/or ligament reconstruction. Although numerous studies have reported improved clinical outcomes with meniscal allograft transplantation, high-quality studies with control groups are lacking in the literature. This article describes the current indications, graft types and sizing, surgical techniques, and a review of the literature with a focus on the role of concomitant procedures. PMID- 19306735 TI - Osteochondral allografts: state of the art. AB - The use of osteochondral allografts to treat focal osteochondral lesions continues to gain popularity, supported by long-term results. Clinicians must be knowledgeable concerning the possible risks of disease transmission, graft rejection, infection, and graft failure to advise the patient and obtain an informed consent. With advancing scientific and clinical research, future operative indications will likely continue to expand. A significant amount of literature regarding storage methods has recently been published; it is hoped that continued research will lead to techniques for prolonged graft storage to prevent availability concerns. PMID- 19306736 TI - Collateral ligament augmentation versus reconstruction using allograft tissue. AB - Injuries to the collateral ligaments of the knee are very common. Recognition of collateral ligament laxity is extremely important, especially when associated with a cruciate ligament injury. Reconstruction of the collaterals are necessary when addressing these combined instabilities. PMID- 19306737 TI - Anatomic double-bundle revision anterior cruciate ligament surgery using fresh frozen allograft tissue. AB - Revision anterior cruciate ligament (ACL) reconstruction presents many technical considerations not seen in primary ACL reconstruction. A variety of allograft options are available for use during revision ACL reconstruction, including bone patella tendon-bone, calcaneus-Achilles tendon, and all soft-tissue grafts. Anatomic double-bundle ACL technique improves knee kinematics and provides the rotatory stability necessary to return to ACL-dependent activities. PMID- 19306738 TI - Future of allografts in sports medicine. AB - Allografts play a prominent role in sports medicine, and their usage has increased dramatically over the past few decades, but the role of allograft in the future of sports medicine largely depends on several factors: (1) the ability of the tissue banking industry to convince both surgeons and the general population that tissue procurement is safe and nearly disease-free, (2) the ability to sterilize tissue with minimal compromise to tissue integrity, (3) successful clinical outcomes with allograft, and (4) the advent of artificial scaffolds and ligaments that function as well. PMID- 19306745 TI - Elective nodal irradiation for locally advanced non-small-cell lung cancer: it's called cancer for a reason. PMID- 19306746 TI - Larynx preservation clinical trial design: key issues and recommendations-a consensus panel summary. AB - PURPOSE: To develop guidelines for the conduct of Phase III clinical trials of larynx preservation in patients with locally advanced laryngeal and hypopharyngeal cancer. METHODS AND MATERIALS: A multidisciplinary international consensus panel developed recommendations after reviewing results from completed Phase III randomized trials, meta-analyses, and published clinical reports with updates available through November, 2007. The guidelines were reviewed and approved by the panel. RESULTS: According to the recommendations, the trial population should include patients with T2 or T3 laryngeal or hypopharyngeal squamous cell carcinoma not considered for partial laryngectomy and exclude those with laryngeal dysfunction or age greater than 70 years. Functional assessments should include speech and swallowing. Voice should be routinely assessed with a simple, validated instrument. The primary endpoint should capture survival and function. The panel created a new endpoint: laryngo-esophageal dysfunction-free survival. Events are death, local relapse, total or partial laryngectomy, tracheotomy at 2 years or later, or feeding tube at 2 years or later. Recommended secondary endpoints are overall survival, progression-free survival, locoregional control, time to tracheotomy, time to laryngectomy, time to discontinuation of feeding tube, and quality of life/patient-reported outcomes. Correlative biomarker studies for near-term trials should include estimated glomerular filtration rate, excision repair cross-complementary-1 gene, E-cadherin and beta catenin, epiregulin and amphiregulin, and TP53 mutation. CONCLUSIONS: Revised trial designs in several key areas are needed to advance the study of larynx preservation. With consistent methodologies, clinical trials can more effectively evaluate and quantify the therapeutic benefit of novel treatment options for patients with locally advanced laryngeal and hypopharyngeal cancer. PMID- 19306748 TI - Does incidental irradiation with doses below 50 gy effectively reduce isolated nodal failures in non-small-cell lung cancer: dose-response relationship. AB - PURPOSE: To evaluate the dose-response relationship for a wide range of doses lower than 50 Gy delivered to the hilar and mediastinal lymph node stations from incidental irradiation in 220 patients with non-small-cell lung cancer (NSCLC) treated with three-dimensional conformal radiotherapy. The endpoint was isolated nodal recurrence (INR) in stations that were initially negative. METHODS AND MATERIALS: The individual responses of 2596 nodal stations were analyzed. Different fractionation schedules were used in different patients. Total prescribed tumor doses ranged from 52 Gy to 74 Gy given over 16-56 days. There were 1198 nodal stations (46%) within and 1398 stations beyond the elective nodal irradiation (ENI) volumes. The INR incidence was estimated for six dose levels ranging from 5 +/- 5 Gy to > or = 56 Gy. RESULTS: There were a total of 25 INRs in 17 patients (8%). The incidence of INR within the electively treated volumes was 0.58%, compared with 1.28% in nodal stations beyond the ENI. Almost 80% of the INRs occurred during 10 months of follow-up. A strong dose-response relationship was seen for the lower "incidental" doses, most of which were less than 50 Gy. As the dose increased from 5 +/- 5 Gy to 40 +/- 5 Gy, the rate of freedom from INR increased from 12% to 76% (p = 0.005). CONCLUSIONS: There is evidence of a dose-response relationship between a reduction in the rate of INR and doses lower than 50 Gy. This suggests that incidental irradiation can eradicate at least some subclinical metastases in regional lymph nodes. PMID- 19306747 TI - Impact of radiotherapy on fertility, pregnancy, and neonatal outcomes in female cancer patients. AB - PURPOSE: Radiation has many potential long-term effects on cancer survivors. Female cancer patients may experience decreased fertility depending on the site irradiated. Oncologists should be aware of these consequences and discuss options for fertility preservation before initiating therapy. METHODS AND MATERIALS: A comprehensive review of the existing literature was conducted. Studies reporting the outcomes for female patients treated with cranio-spinal, abdominal, or pelvic radiation reporting fertility, pregnancy, or neonatal-related outcomes were reviewed. RESULTS: Cranio-spinal irradiation elicited significant hormonal changes in women that affected their ability to become pregnant later in life. Women treated with abdomino-pelvic radiation have an increased rate of uterine dysfunction leading to miscarriage, preterm labor, low birth weight, and placental abnormalities. Early menopause results from low-dose ovarian radiation. Ovarian transposition may decrease the rates of ovarian dysfunction. CONCLUSIONS: There is a dose-dependent relationship between ovarian radiation therapy (RT) and premature menopause. Patients treated with RT must be aware of the impact of treatment on fertility and explore appropriate options. PMID- 19306749 TI - Sensitivity analysis of parameters in linear-quadratic radiobiologic modeling. AB - PURPOSE: Radiobiologic modeling is increasingly used to estimate the effects of altered treatment plans, especially for dose escalation. The present article shows how much the linear-quadratic (LQ) (calculated biologically equivalent dose [BED] varies when individual parameters of the LQ formula are varied by +/-20% and by 1%. METHODS: Equivalent total doses (EQD2 = normalized total doses (NTD) in 2-Gy fractions for tumor control, acute mucosal reactions, and late complications were calculated using the linear- quadratic formula with overall time: BED = nd (1 + d/ [alpha/beta]) - log(e)2 (T - Tk) / alphaTp, where BED is BED = total dose x relative effectiveness (RE = nd (1 + d/ [alpha/beta]). Each of the five biologic parameters in turn was altered by +/-10%, and the altered EQD2s tabulated; the difference was finally divided by 20. EQD2 or NTD is obtained by dividing BED by the RE for 2-Gy fractions, using the appropriate alpha/beta ratio. RESULTS: Variations in tumor and acute mucosal EQD ranged from 0.1% to 0.45% per 1% change in each parameter for conventional schedules, the largest variation being caused by overall time. Variations in "late" EQD were 0.4% to 0.6% per 1% change in the only biologic parameter, the alpha/beta ratio. For stereotactic body radiotherapy schedules, variations were larger, up to 0.6 to 0.9 for tumor and 1.6% to 1.9% for late, per 1% change in parameter. CONCLUSIONS: Robustness occurs similar to that of equivalent uniform dose (EUD), for the same reasons. Total dose, dose per fraction, and dose-rate cause their major effects, as well known. PMID- 19306750 TI - Use of the concept of equivalent biologically effective dose (BED) to quantify the contribution of hyperthermia to local tumor control in radiohyperthermia cervical cancer trials, and comparison with radiochemotherapy results. AB - PURPOSE: To express the magnitude of contribution of hyperthermia to local tumor control in radiohyperthermia (RT/HT) cervical cancer trials, in terms of the radiation-equivalent biologically effective dose (BED) and to explore the potential of the combined modalities in the treatment of this neoplasm. MATERIALS AND METHODS: Local control rates of both arms of each study (RT vs. RT+HT) reported from randomized controlled trials (RCT) on concurrent RT/HT for cervical cancer were reviewed. By comparing the two tumor control probabilities (TCPs) from each study, we calculated the HT-related log cell-kill and then expressed it in terms of the number of 2 Gy fraction equivalents, for a range of tumor volumes and radiosensitivities. We have compared the contribution of each modality and made some exploratory calculations on the TCPs that might be expected from a combined trimodality treatment (RT+CT+HT). RESULTS: The HT-equivalent number of 2 Gy fractions ranges from 0.6 to 4.8 depending on radiosensitivity. Opportunities for clinically detectable improvement by the addition of HT are only available in tumors with an alpha value in the approximate range of 0.22-0.28 Gy(-1). A combined treatment (RT+CT+HT) is not expected to improve prognosis in radioresistant tumors. CONCLUSION: The most significant improvements in TCP, which may result from the combination of RT/CT/HT for locally advanced cervical carcinomas, are likely to be limited only to those patients with tumors of relatively low-intermediate radiosensitivity. PMID- 19306751 TI - Comparison of biological effectiveness of carbon-ion beams in Japan and Germany. AB - PURPOSE: To compare the biological effectiveness of 290 MeV/amu carbon-ion beams in Chiba, Japan and in Darmstadt, Germany, given that different methods for beam delivery are used for each. METHODS AND MATERIALS: Murine small intestine and human salivary gland tumor (HSG) cells exponentially growing in vitro were irradiated with 6-cm width of spread-out Bragg peaks (SOBPs) adjusted to achieve nearly identical beam depth-dose profiles at the Heavy-Ion Medical Accelerator in Chiba, and the SchwerIonen Synchrotron in Darmstadt. Cell kill efficiencies of carbon ions were measured by colony formation for HSG cells and jejunum crypts survival in mice. Cobalt-60 gamma rays were used as the reference radiation. Isoeffective doses at given survivals were used for relative biological effectiveness (RBE) calculations and interinstitutional comparisons. RESULTS: Isoeffective D(10) doses (mean +/- standard deviation) of HSG cells ranged from 2.37 +/- 0.14 Gy to 3.47 +/- 0.19 Gy for Chiba and from 2.31 +/- 0.11 Gy to 3.66 +/- 0.17 Gy for Darmstadt. Isoeffective D(10) doses of gut crypts after single doses ranged from 8.25 +/- 0.17 Gy to 10.32 +/- 0.14 Gy for Chiba and from 8.27 +/- 0.10 Gy to 10.27 +/- 0.27 Gy for Darmstadt, whereas isoeffective D(30) doses after three fractionated doses were 9.89 +/- 0.17 Gy through 13.70 +/- 0.54 Gy and 10.14 +/- 0.20 Gy through 13.30 +/- 0.41 Gy for Chiba and Darmstadt, respectively. Overall difference of RBE between the two facilities was 0-5% or 3 7% for gut crypt survival or HSG cell kill, respectively. CONCLUSION: The carbon ion beams at the National Institute of Radiological Sciences in Chiba, Japan and the Gesellschaft fur Schwerionenforschung in Darmstadt, Germany are biologically identical after single and daily fractionated irradiation. PMID- 19306752 TI - Studies on pentoxifylline and tocopherol combination for radiation-induced heart disease in rats. AB - PURPOSE: To investigate whether the application of pentoxifylline (PTX) and tocopherol l (Vit. E) could modify the development of radiation-induced heart disease and downregulate the expression of transforming growth factor (TGF) beta1mRNA in rats. METHODS AND MATERIALS: A total of 120 Sprague-Dawley rats were separated into four groups: control group, irradiated group, experimental group 1, and experiment group 2. Supplementation was started 3 days before irradiation; in experimental group 1, injection of PTX (15 mg/kg/d) and Vit. E (5.5 mg/kg/d) continued till the 12th week postirradiation, whereas in experimental group 2 it was continued until the 24th week postirradiation. All rats were administrated a single dose of 20 Gy irradiation to the heart except the control group. Histopathologic evaluation was performed at various time points (Days 1, 2, 4, 8, and 12 and 24th week) up to 24 weeks after irradiation. Changes of levels of TGF beta1 mRNA expression were also investigated at the same time points using competitive polymerase chain reaction. RESULTS: Compared with the irradiated group, levels of TGF-beta1 mRNA of the rat hearts were relatively low in the two experimental groups on the 12th week postirradiation. In experimental group 1, there was a rebound expression of TGF-beta1 mRNA on the 24th week postirradiation, whereas that of the experimental group 2 remained low (p < 0.05). The proportions of collagen fibers of the two experimental groups were lower than that of irradiated group (p < 0.05). A rebound could be observed in the experimental group 1. CONCLUSION: PTX and Vit. E downregulated the expression of TGF-beta1 mRNA. The irradiated rat hearts showed a marked pathologic response to the drugs. The withdrawal of drugs in the 12th week postirradiation could cause rebound effects of the development of fibrosis. PMID- 19306753 TI - Setup uncertainties of anatomical sub-regions in head-and-neck cancer patients after offline CBCT guidance. AB - PURPOSE: To quantify local geometrical uncertainties in anatomical sub-regions during radiotherapy for head-and-neck cancer patients. METHODS AND MATERIALS: Local setup accuracy was analyzed for 38 patients, who had received intensity modulated radiotherapy and were regularly scanned during treatment with cone beam computed tomography (CBCT) for offline patient setup correction. In addition to the clinically used large region of interest (ROI), we defined eight ROIs in the planning CT that contained rigid bony structures: the mandible, larynx, jugular notch, occiput bone, vertebrae C1-C3, C3-C5, and C5-C7, and the vertebrae caudal of C7. By local rigid registration to successive CBCT scans, the local setup accuracy of each ROI was determined and compared with the overall setup error assessed with the large ROI. Deformations were distinguished from rigid body movements by expressing movement relative to a reference ROI (vertebrae C1-C3). RESULTS: The offline patient setup correction protocol using the large ROI resulted in residual systematic errors (1 SD) within 1.2 mm and random errors within 1.5 mm for each direction. Local setup errors were larger, ranging from 1.1 to 3.4 mm (systematic) and 1.3 to 2.5 mm (random). Systematic deformations ranged from 0.4 mm near the reference C1-C3 to 3.8 mm for the larynx. Random deformations ranged from 0.5 to 3.6 mm. CONCLUSION: Head-and-neck cancer patients show considerable local setup variations, exceeding residual global patient setup uncertainty in an offline correction protocol. Current planning target volume margins may be inadequate to account for these uncertainties. We propose registration of multiple ROIs to drive correction protocols and adaptive radiotherapy to reduce the impact of local setup variations. PMID- 19306754 TI - A study on target positioning error and its impact on dose variation in image guided stereotactic body radiotherapy for the spine. AB - PURPOSE: To investigate the amount of target positioning error and evaluate its dosimetric impact during image-guided stereotactic body radiotherapy for single fraction spine treatment. METHODS AND MATERIALS: A prescription dose of 15 Gy and five to nine coplanar intensity-modulated beams were used. The patient was immobilized with a custom-fit vacuum mold, and the target was localized with a volumetric cone-beam CT image. A robotic couch with six degrees of freedom was used for target adjustment. For evaluation a cone-beam CT image was obtained at the end of treatment. Both target positioning error and its dosimetric impact were investigated for the first 9 cases. RESULTS: For cases studied, translational errors were 0.9 +/- 0.5 mm (lateral), 1.2 +/- 0.9 mm (longitudinal), 0.7 +/- 0.6 mm (vertical), and 1.8 +/- 1.0 mm (vector), and rotational errors were 1.6 degrees +/- 1.3 degrees (pitch), 0.8 degrees +/- 0.9 degrees (roll), and 0.8 degrees +/- 0.4 degrees (yaw). For the clinical target volume, D(95) (dose to 95% of target volume), D(90), D(max), and D(mean) were evaluated. Only 1 case showed significant dose variations, reaching up to 18% in D(95). The spinal cord dose was evaluated by observing D(0.1) (dose to 0.1 cm(3)), D(0.5), D(1.0), and D(max). Although 1 case showed a dose change reaching up to 30% in D(max), cord dose was within the planning tolerance limit in all but 2 cases (3% higher in one and 0.4% higher in the other). CONCLUSION: The implemented image-guided stereotactic body radiotherapy provides precise target localization. However, despite reasonably precise spatial precision, dosimetric perturbation can be significant because of both extremely steep dose gradients and close distances between the target and the spinal cord. PMID- 19306755 TI - 3D in vivo dosimetry using megavoltage cone-beam CT and EPID dosimetry. AB - PURPOSE: To develop a method that reconstructs, independently of previous (planning) information, the dose delivered to patients by combining in-room imaging with transit dose measurements during treatment. METHODS AND MATERIALS: A megavoltage cone-beam CT scan of the patient anatomy was acquired with the patient in treatment position. During treatment, delivered fields were measured behind the patient with an electronic portal imaging device. The dose information in these images was back-projected through the cone-beam CT scan and used for Monte Carlo simulation of the dose distribution inside the cone-beam CT scan. Validation was performed using various phantoms for conformal and IMRT plans. Clinical applicability is shown for a head-and-neck cancer patient treated with IMRT. RESULTS: For single IMRT beams and a seven-field IMRT step-and-shoot plan, the dose distribution was reconstructed within 3%/3mm compared with the measured or planned dose. A three-dimensional conformal plan, verified using eight point dose measurements, resulted in a difference of 1.3 +/- 3.3% (1 SD) compared with the reconstructed dose. For the patient case, planned and reconstructed dose distribution was within 3%/3mm for about 95% of the points within the 20% isodose line. Reconstructed mean dose values, obtained from dose-volume histograms, were within 3% of prescribed values for target volumes and normal tissues. CONCLUSIONS: We present a new method that verifies the dose delivered to a patient by combining in-room imaging with the transit dose measured during treatment. This verification procedure opens possibilities for offline adaptive radiotherapy and dose-guided radiotherapy strategies taking into account the dose distribution delivered during treatment sessions. PMID- 19306756 TI - A rabbit irradiation platform for outcome assessment of lung stereotactic radiosurgery. AB - PURPOSE: To evaluate a helical tomotherapy-based rodent radiosurgery platform that reproduces human image-guided radiosurgery treatment to study radiobiologic effects of stereotactic radiosurgery on lung tissues using functional magnetic resonance imaging (MRI). METHODS AND MATERIALS: Hypofractionated radisourgery (20 Gy x 3) was delivered to the right lung of three New Zealand rabbits using Helical TomoTherapy with MVCT image guidance. Contrast-enhanced MR perfusion, hyperpolarized helium-3 MR ventilation, and CT were obtained before radiation and monthly for 4 months after radiation. All MRI was performed on a 1.5-T whole-body scanner with broad-band capabilities. RESULTS: Precise dose delivery to 1.6 cc of the lower right lung was achieved without additional immobilization. No deficits were detected at baseline with respect to perfusion and ventilation. Lung perfusion deficits in the irradiated lung regions began at 2 months after radiation and worsened with time. No ventilation deficits were observed after radiation. Decrease in lung CT density in irradiated regions was observed after radiation, but the changes were less significant than those in perfusion MRI. CONCLUSIONS: We demonstrated that highly conformal radiation can be reproducibly delivered to a small volume of rodent lung on a widely available clinical unit. The radiation-induced lung injury can be detected as early as 2 months after radiation with perfusion MRI. The primary pattern of injury agrees with previously reported endothelial damage to radiosurgical radiation doses. This experimental design provides a cost-effective methodology for producing radiosurgical injuries in rodents that reproduces current human treatments for studying radiation injury and agents that might affect it. PMID- 19306757 TI - Influence of rotations on dose distributions in spinal stereotactic body radiotherapy (SBRT). AB - PURPOSE: To evaluate the impact of rotational setup errors on dose distribution in spinal stereotactic body radiotherapy (SBRT). METHODS AND MATERIALS: Thirty nine cone beam computed tomography (CBCT) scans from 16 SBRT treatment courses were analyzed. Alignment (including rotation) to the treatment planning computed tomography was performed, followed by translational alignment that reproduced the actual positioning. The planned fluence was then applied to determine the delivered dose to the targets and organs at risk. RESULTS: The mean planning target volume (PTV) was 71.01 mL (SD +/- 60.05; range, 22.62-250.65 mL). Prescribed dose (to the 62-82% isodose) was 14-30 Gy in one to six fractions. The average rotational displacements were 0.38 +/- 1.21, 1.12 +/- 1.82, and -0.51 +/- 2.0 degrees with maximal rotations of -4.29, 5.76, and -6.64 degrees along the x (pitch), y (yaw), and z (roll) axes, respectively. PTV coverage changed by an average of -0.07 Gy (SD +/- 0.20 Gy) between the rotated and the original plan, representing 0.92% of prescription dose (SD +/- 2.65%). For the spinal cord, planned with 2-mm expansion to create a planning organ at risk volume (PRV), the difference in minimum dose to the upper 10% of the PRV volume was 0.03 +/- 0.3 Gy (maximum, 0.9 Gy). Other organs at risk saw insignificant changes in dose. CONCLUSIONS: PRV expansion generally assures safe treatment delivery in the face of typically encountered rotations. Given the variability of delivered dose within this expansion for certain cases, caution should be taken to properly interpret doses to the cord when considering clinical dose limits. PMID- 19306759 TI - Re: what do recent studies on lumpectomy cavity volume change imply for breast clinical target volumes? PMID- 19306760 TI - Thyroid cancer as second primary malignancy in breast cancer patients. PMID- 19306761 TI - Elective nodal irradiation (ENI) does not appear to provide a clear benefit for patients with unresectable non-small-cell lung cancer (NSCLC): in regard to Schild et al. (Int J Radiat Oncol Biol Phys 2008;72:335-342). PMID- 19306764 TI - The nuclear technique most frequently used by cardiologists is nuclear perfusion imaging, which is a fairly mature technology. Foreword. PMID- 19306765 TI - Preface: The changing face of nuclear cardiology. PMID- 19306766 TI - New trends in camera and software technology in nuclear cardiology. AB - This article describes advancements in hardware and software for myocardial perfusion imaging that are becoming commercialized today and their implication in clinical practice. PMID- 19306767 TI - Cardiac positron emission tomography: current clinical practice. AB - In the last two decades, the field of nuclear cardiology has experienced significant progress. The introduction of positron emission tomography (PET) imaging represented a major breakthrough that has significantly contributed to a better understanding of physiology and pathophysiology of several heart diseases. Currently, PET imaging is recognized as a well-established method to assess cardiac perfusion, function, metabolism, and viability. This article summarizes the main clinical applications of state-of-the art cardiac PET technology. PMID- 19306768 TI - Hybrid imaging: integration of nuclear imaging and cardiac CT. AB - The integration of nuclear medicine cameras with multidetector CT scanners provides a unique opportunity to delineate cardiac and vascular anatomic abnormalities and their physiologic consequences in a single setting. By revealing the burden of anatomic coronary artery disease and its physiologic significance, hybrid imaging can provide unique information that may improve noninvasive diagnosis, risk assessment, and management of coronary artery disease. By integrating the detailed anatomic information from CT with the high sensitivity of radionuclide imaging to evaluate targeted molecular and cellular abnormalities, hybrid imaging may play a key role in shaping the future of molecular diagnostics and therapeutics. This article reviews potential clinical applications of hybrid imaging in cardiovascular disease. PMID- 19306769 TI - Nuclear imaging in heart failure. AB - Heart failure is becoming the main clinical challenge in cardiology in the twenty first century and is associated with high morbidity and mortality. Currently, several therapeutic options are available for heart failure patients, including medical therapy, revascularization, advanced cardiac surgery, device therapy, and cardiac transplantation. Future therapies are directed at cell and gene therapy. In this article the role of nuclear imaging in the management of heart failure patients is discussed. PMID- 19306770 TI - Quantification of myocardial blood flow: what is the clinical role? AB - Quantification of regional myocardial blood flow and of its responses to targeted physiologic and pharmacologic interventions, which is now available with positron emitting tracers of blood flow and positron emission tomography (PET), extends the diagnostic potential of standard myocardial perfusion imaging. These noninvasive flow measurements serve as tools for quantifying functional consequences of epicardial coronary artery disease, as well as of impairments in microcirculatory reactivity that escape detection by standard perfusion imaging. Flow measurements are clinically useful for more comprehensively assessing the extent and severity of coronary vascular disease or impairments in microcirculatory function in noncoronary cardiac disease. Flow estimates in these disorders contain independent or unique prognostic information about future major cardiac events. Flow measurements are also useful for assessing the coronary risk, for predicting long-term cardiovascular events, and for monitoring the effectiveness of risk reduction strategies. PMID- 19306771 TI - Translation of myocardial metabolic imaging concepts into the clinics. AB - Flexibility in myocardial substrate metabolism for energy production is fundamental to cardiac health. This loss in plasticity or flexibility leads to overdependence on the metabolism of an individual category of substrates, with the predominance in fatty acid metabolism characteristic of diabetic heart disease and the accelerated glucose use associated with pressure-overload left ventricular hypertrophy being prime examples. There is a strong demand for accurate noninvasive imaging approaches of myocardial substrate metabolism that can facilitate the crosstalk between the bench and the bedside, leading to improved patient management paradigms. In this article potential future applications of metabolic imaging, particularly radionuclide approaches, for assessment of cardiovascular disease are discussed. PMID- 19306772 TI - Cardiac neuronal imaging at the edge of clinical application. AB - Cardiac neuronal innervation plays an important role in normal cardiac function and is adversely affected in the presence of disease. In particular, radiotracer imaging of cardiac sympathetic function has been extensively investigated and not only provides a method of assessing the severity of disease but also has repeatedly been shown to be prognostically useful with a potential for helping to guide patient management. SPECT imaging of myocardial uptake of (123)I-mIBG, an analog of the sympathetic neurotransmitter norepinephrine, has been the most studied, but PET neurotracers, such as (11)C-HED, are also under investigation. The ability of cardiac neuronal imaging to visualize and measure underlying molecular processes should allow it to provide a perspective on cardiac disease that other testing modalities cannot. PMID- 19306774 TI - Radiotracer imaging of atherosclerotic plaque biology. AB - Traditional imaging modalities used in the assessment of atherosclerotic plaque have focused on anatomic characteristics of size, location and luminal encroachment. The ability to identify plaques that are at risk for rupture, and thus may go on to cause clinical events, remains limited, however. By labeling tracer compounds capable of identifying important cellular or molecular processes involved in plaque vulnerability with radioactive isotopes, there is now potential for the noninvasive identification of vulnerable plaques. This article discusses several radiotracers that can report on high-risk plaque pathophysiology. PMID- 19306773 TI - New molecular imaging targets to characterize myocardial biology. AB - Molecular imaging represents a targeted approach to noninvasively assess biologic (both physiologic and pathologic) processes in vivo. Ideally the goal of molecular imaging is not just to provide diagnostic and prognostic information based on identification of the molecular events associated with a pathologic process but rather to guide individually tailored pharmacologic, cell-based, or genetic therapeutic regimens. This article reviews the recent advances in myocardial molecular imaging in the context of the cardiovascular processes of angiogenesis, apoptosis, inflammation, and ventricular remodeling. The focus is on radiotracer-based single photon emission computed tomography and positron emission tomography molecular imaging approaches. PMID- 19306775 TI - Role of nuclear imaging in regenerative cardiology. AB - Advances in noninvasive imaging techniques may aid in the understanding of cardiac stem cell therapy. Nuclear imaging enables in vivo evaluation of myocardial perfusion, metabolism, and function, in addition to the stem cell fate. This article summarizes recent clinical and experimental nuclear imaging studies in cardiac stem cell therapy. PMID- 19306776 TI - [Obstructive lung diseases in a French prison: results of systematic screening]. AB - BACKGROUND: French prisoners have health problems that have been inadequately treated before imprisonment. This population has insufficient access to the healthcare system. Addictive behaviours, particularly smoking, are widespread. The aim of the study is to evaluate the prevalence of airflow limitation by using a primary care screening method adapted for the correctional facility and its inmates. METHOD: The screening of airflow limitation using a mobile spirometer is carried out in inmates consulting the primary care unit (UCSA) of Amiens prison. Patients consulting the UCSA between 16 August and 17 October 2006 and providing their consent are included in the study. The criteria for exclusion are: a counter-indication for spirometry, poor compliance with the effort of forced expiry after eight efforts, as well as refusal to take part in the study. The descriptive statistical analysis includes all of the quantitative and qualitative variables. RESULTS: Among the 210 patients included in the sample, only five patients refused to take part in the study. Their mean age was 37 (range: 16-65) and 90% were men. Ninety percent of this population were active smokers. Sixty percent of these smokers would like to quit. The spirometry detected 11% undiagnosed airflow limitation: 11 prisoners suffered from chronic obstructive lung disease and 13 prisoners suffered from asthma. DISCUSSION: Given the relative youth and high risk nature of these diagnosed patients, the potential for the long or short term aggravation, and a growing recognition of the seriousness of exposure to tobacco, the authors suggest that the systematic screening of inmates for airflow limitation may be used to assist in detecting serious health issues. Along with new French antismoking legislation, this screening may enable primary care workers to better reduce smoking habits in prisons. PMID- 19306777 TI - [Varicella pneumonia: a serious complication of varicella in adults]. AB - Pneumonia caused by varicella infection is a serious and severe complication of the disease when it occurs in adults. The authors report two cases of a varicella pneumonia in a 36 and a 41 year-old non-immunocompromised man. They were admitted to hospital for acute respiratory failure, not requiring intensive care, associated with exanthema. High resolution CT revealed well-defined diffuse nodules throughout both lungs, nodules with a surrounding halo of ground-glass opacity. Thrombocytopenia, hepatic cytolysis and an increase in lacticodeshydrogenase were noted. The patients made a full recovery with anti viral therapy and oxygen therapy. Varicella pneumonia is the most common complication of varicella in adults. Anti-viral treatment is usually successful. PMID- 19306778 TI - [Shoulder pain revealing tuberculosis of the humerus]. AB - Tuberculous spondylitis is the most common form of musculoskeletal tuberculosis. However, extraspinal manifestations have been described with tuberculosis of the wrist, femur, foot or shoulder, as in the patient presented. Because of an often indolent clinical presentation, the diagnosis is delayed and antituberculous treatment is not able to prevent serious bone destruction. PMID- 19306779 TI - [Erasmus syndrome: case report]. AB - Erasmus' syndrome involves the association of systemic scleroderma and exposure to silica particles with or without silicosis. The authors report the observation of a miner with a history of professional silicosis resulting in pulmonary fibrosis. Twenty-five years later, the patient consulted for dysphagia associated with inflammatory arthralgia, cutaneous sclerosis extended to the face and limbs, sclerodactyly and Raynaud's phenomenon. The diagnosis of scleroderma was confirmed and Erasmus' syndrome was retained. PMID- 19306780 TI - [A late post-traumatic diaphragmatic hernia revealed by a tension fecopneumothorax (a case report)]. AB - INTRODUCTION: Post-traumatic diaphragmatic hernia is a particular lesion in traumatology that may be neglected. Thus, the diagnosis may be delayed for a few days to several months and only be made following a complication. The left diaphragmatic cupola is the most touched. Tension fecopneumothorax following diaphragmatic hernia perforation in the pleural cavity is a rare but particularly severe complication of traumatic diaphragmatic hernia. CASE REPORT: A 68-year-old man was admitted for acute intestinal occlusion with respiratory distress. A history of a violent blunt thoraco-abdominal traumatism resulting from a traffic accident eight years before was noted. The chest x-ray revealed an abundant hydropneumothorax and the thoracic scan revealed abundant effusion with heterogeneous density in the left pleural cavity, associated with an intrapleural hernia of the large intestine. An emergency thoracolaparotomy discovered tension fecopneumothorax secondary to intrathoracic perforation of the transverse colon through a left hemidiaphragm defect. The surgical treatment consisted of hernia reduction, pleural drainage, colostomy and repair of the diaphragmatic defect. CONCLUSION: The possibility of diaphragmatic hernia should be kept in mind in case of violent blunt thoraco-abdominal traumatism or basithoracic wound. In this way, complications such as tension fecopneumothorax that could threaten the functional and vital prognosis may be prevented. PMID- 19306781 TI - [Thyroid metastasis of lung cancer and abnormal thyroid function--a case report]. AB - The thyroid gland is a very rare location of metastasis and the metastatic involvement of the thyroid is mostly asymptomatic. The authors report one of the first cases of pulmonary adenocarcinoma associated with painful metastatic involvement of the thyroid gland. Temporary hyperthyroidism was noted, followed, two months later, by clinically and biologically proven hypothyroidism with positive antithyroglobulin antibodies. The suspect goiter was detected by diffuse hyperfixation on 18-FDG PET Scan and the ultrasonography revealed two hypoechogenic nodules. The fine needle biopsy confirmed the metastatic origin of these nodules. The evolution after five cycles of chemotherapy by cisplatine and docetaxel was marked by a complete regression of the thyroid metastasis and an improvement in the thyroid function. PMID- 19306782 TI - [An unusual paraneoplastic manifestation in lung cancer: eosinophilic erythroderma]. AB - An 81-year-old man was admitted for generalized weakness, erythrodermia and eosinophilia. His chest CT showed nodules related to lung adenocarcinoma. Chemotherapy induced a tumour response with the disappearance of the erythrodermia and eosinophilia. A tumour relapse indicating the recurrence of the erythrodermia and eosinophilia was confirmed 2 months after completion of the chemotherapy. The outcome was rapidly fatal. The evolution of the symptoms suggests that eosinophilic erythrodermia is a paraneoplastic syndrome. Cutaneous paraneoplastic syndromes are rare but may be associated with lung cancer. PMID- 19306783 TI - [Endobronchial lipoma: imaging findings. A case report]. AB - Endobronchial lipoma is a rare benign bronchial tumour. A search should be carried out on submillimetre MDCT scan slices in patients presenting segmental or lobar collapse or recurrent pulmonary infection in the same bronchial territories. The authors report MDCT and MR imaging in a patient with endobronchial lipoma discovered on an MDCT scan. PMID- 19306784 TI - [Perfusion and diffusion-weighted MR imaging in the early staging and the follow up of patients with lung cancer]. AB - Tissue characterization is a major and ultimate goal of imaging, whether morphological (Computed Tomography, Magnetic Resonance Imaging) or metabolic (PET FDG-[18F]). Functional imaging, using the MRI, began several years ago with the perfusion of lung nodules and very recently with diffusion-weighted imaging applied to the lung cancer. The authors review the interest and the place of diffusion-weighted and perfusion MR imaging in the diagnosis, early staging and follow-up of patients with lung cancer. PMID- 19306785 TI - [Contribution of PET/CT for staging and prognosis of non-small cell lung carcinoma. Assessment of the pathological response]. PMID- 19306786 TI - [Splenothoracic hydatidosis: case report]. PMID- 19306787 TI - Fumbling the handoff: managing the transition to adult care for adolescents with chronic conditions. PMID- 19306788 TI - Reactive arthritis: a review. AB - This review article summarizes the available literature on adolescent reactive arthritis. A review of the pathophysiology, diagnosis, and treatment guidelines will be helpful to better diagnose and treat reactive arthritis. PMID- 19306789 TI - Expectations and experiences of adolescents with congenital heart disease on being transferred from pediatric cardiology to an adult congenital heart disease program. AB - PURPOSE: To investigate the expectations and experiences of adolescents on transferring from a pediatric cardiology program to an adult congenital heart disease program. METHODS: A qualitative, phenomenologic study was conducted, in which semi-structured, in-depth interviews were performed with 14 adolescents (aged 15-17 years) with congenital heart disease. Interviews were tape-recorded and transcribed verbatim. Data were analyzed using procedures described by Colaizzi. RESULTS: The study identified six themes that characterized adolescents' views of the transfer process: leaving pediatric cardiology was viewed as normal; leaving behind familiar surroundings; a positive wait-and-see attitude toward the adult congenital heart disease program; adjusting to a new environment; a need for better information; and a shift in roles between the adolescent and his parents. CONCLUSION: Although adolescents with congenital heart disease have a positive attitude toward transferring to an adult congenital heart disease program, they identified aspects of the transfer that healthcare professionals need to address. Patients and their families should be prepared for the transfer by informing them about the adult program and its healthcare providers. A formal transition program can play a role in this respect. PMID- 19306790 TI - Decision-making authority and substance abuse treatment for adolescents: a survey of state laws. AB - PURPOSE: State laws concerning decision-making authority for voluntary inpatient substance abuse (SA) treatment of minors may be a potential barrier to appropriate treatment. We sought to identify and classify relevant laws related to the provision of voluntary inpatient SA treatment to adolescents 12 to 17 years (minors) as an exploratory assessment to improve understanding of how these laws might affect treatment decisions. METHODS: In summer 2006, we conducted a survey of statutes, regulations, and legal cases in the 50 states and the District of Columbia regarding the authority of parents (or guardians) and minors to make treatment decisions for voluntary inpatient SA treatment. RESULTS: All 50 states have laws applicable to voluntary inpatient SA treatment for adolescents, and the laws vary significantly throughout the nation. If a minor and parent disagree about SA treatment, some states defer to the decision-making authority of the minor, whereas other states defer to the parent. Most significantly, the majority of states fail to specify whether the minor's or the parent's decision will control in the event of a conflict. CONCLUSIONS: The lack of clarity in state laws regarding decision-making authority for voluntary inpatient SA treatment of minors may create a potential barrier to treatment for adolescents, especially those with more serious SA problems. This lack of clarity could lead to confusion among parents, adolescents, healthcare professionals, and treatment facilities, and ultimately could result in a failure to treat adolescents in need of medical attention. Policymakers should ensure that state laws clearly specify procedures to enable treatment if a conflict arises between adolescents and parents, including procedures to ensure that the due process rights of adolescents are protected. PMID- 19306791 TI - Adolescent alcohol use, suicidal ideation, and suicide attempts. AB - PURPOSE: To examine the association between self-reported alcohol use and suicide attempts among adolescents who did and did not report suicidal ideation during the past year. METHODS: Screening data from 31,953 students attending schools in the United States that implemented the Signs of Suicide (SOS) program in 2001 2002 were used in this analysis. Two types of alcohol use were investigated: heavy episodic drinking, and drinking while down. Self-reported suicide attempts were regressed on suicidal ideation and both measures of alcohol use, controlling for participants' levels of depressive symptoms, and demographic characteristics. RESULTS: Logistic regression analyses indicated that both drinking while down and heavy episodic drinking were significantly associated with self-reported suicide attempts. Analyses examining the conditional association of alcohol use and suicidal ideation with self-reported suicide attempts revealed that drinking while down was associated with significantly greater risk of suicide attempt among those not reporting suicidal ideation in the past year. Heavy episodic drinking was associated with increased risk of suicide attempt equally among those who did and did not report suicidal ideation. CONCLUSIONS: This study identified the use of alcohol while sad or depressed as a marker for suicidal behavior in adolescents who did not report ideating prior to an attempt, and hence, may not be detected by current strategies for assessing suicide risk. Findings from this study should provide further impetus for alcohol screening among clinicians beyond that motivated by concerns about alcohol and substance use. PMID- 19306792 TI - Influence of subjective social status on the relationship between positive outcome expectations and experimentation with cigarettes. AB - PURPOSE: In Texas, Mexican American (MA) adolescents, and in particular boys, are at increased risk for experimenting with cigarettes compared with their black or white counterparts. Positive outcome expectations (POE), that is, the functional social significance ascribed to cigarettes, and subjective social status (SSS), that is, the adolescents' subjective views of where they lie in the school-based social hierarchy, are independent predictors of smoking. The goal of this study was to test the hypothesis that SSS moderates the relationship between POE and experimentation with cigarettes. METHODS: Moderating effects of SSS were examined using a between-subjects, 2 x 2 analysis of variance and unconditional logistic regression analyses. Using a prospective study design, we followed 1142 MA adolescents aged 11-13 years. Participants completed a baseline survey at home, which assessed POE, SSS, and smoking and were followed via telephone at 6-month intervals over a 12-month period to assess changes in smoking behavior. RESULTS: At follow-up, there were 99 new experimenters. Consistent with our hypothesis, adolescents who reported moderate-low SSS and who held POE at baseline were more likely to have experimented with cigarettes at either follow-up evaluation than their peers with moderate-low SSS who held less POE (odds ratio [OR], 1.92, 95% confidence interval [95% CI], 1.02-3.58). There was no association between outcome expectations and experimenting among adolescents with high SSS (OR, 1.79, CI, .73-4.36). Low SSS boys were more likely to experiment than girls and high SSS boys. CONCLUSIONS: The results of this study indicate that adolescents with moderate-low SSS hold different outcome expectations about smoking than their higher SSS peers. The results underscore the possibility that moderate-low SSS adolescents view behaviors such as smoking as a way to achieve higher SSS and thereby increase their peer social standing. Our results suggest that, in addition to tailoring intervention efforts by gender, placing adolescents of similar social standing to one another within the school into intervention groups that are led by a peer-nominated peer may increase the overall effectiveness of these peer-led prevention efforts. PMID- 19306793 TI - Attitudes and beliefs about adolescent work and workplace safety among parents of working adolescents. AB - PURPOSE: This study was designed to examine the attitudes and beliefs of the parents of working adolescents related to the safety of the employment of their children. METHODS: A cross-sectional telephone survey was conducted in 2003 among English-speaking parents of working adolescents aged 14-18 years in the continental United States. Questions addressed parental concerns about adolescents' employment and potential safety issues at work. RESULTS: Most parents expressed favorable attitudes about adolescent employment, although many expressed concerns about fatigue (48%), problems completing schoolwork (33%), or spending time with families (35%). Half of all parents indicated concerns about adolescents being present during a robbery, and 40-50% were concerned about adolescents working alone or not having adequate safety training. Parents were favorable to laws that regulate the teen work environment, although most (69%) also indicated that parents, not laws, should determine the work that teens do. CONCLUSIONS: Parents of working teens are favorable to their teens working, but they do have varied concerns about safety. Although many parents believe they should set the rules about teen work, they are also generally supportive of governmental regulation of hours and tasks. Helping parents understand child labor policies and consider evidence about work hazards may facilitate their ability to provide appropriate advice to their children. PMID- 19306794 TI - To say or not to say: a qualitative study on the disclosure of their condition by human immunodeficiency virus-positive adolescents. AB - PURPOSE: Human immunodeficiency virus (HIV)-positive adolescents face a number of challenges in dealing with their disease, treatment, and developmental tasks. This qualitative study describes some of the reasons why, and the extent to which, adolescents may or may not disclose their condition to others. METHODS: A semistructured interview lasting 40-110 minutes was conducted with each of 29 adolescents 12-20 years old, 22 female and seven male) living in Switzerland. Interviews were tape recorded and transcribed verbatim. The analysis of the content of interviews allowed us to identify salient topics (e.g., disclosure), which were then explored in detail. RESULTS: Of 29 participants, eight had not disclosed their condition to anyone outside the family, 19 had disclosed it to good friends, and 16 had disclosed it to some teachers. Four participants had engaged in public disclosure, and six of 10 sexually active teenagers disclosed their status to their partners. The attitudes toward disclosure among younger adolescents were mostly related to those of the parents, particularly the mother. Older adolescents, engaged in their search for autonomy, tended to decide independently what to say and to whom. Although foster/adoptive parents would often encourage disclosure, biological parents, especially HIV-positive mothers, insisted on not disclosing the adolescent's status for fear of stigma. CONCLUSION: The health care team should systematically address the issue of disclosure with the adolescent and his family (or foster parents), the aim being to balance the right of the adolescent and that adolescent's family to maintain privacy against the concerns of sexual partners, as well as the adolescent's interest in divulging HIV status to relatives, school staff, and friends. PMID- 19306796 TI - Trends in sexual risk behaviors, by nonsexual risk behavior involvement, U.S. high school students, 1991-2007. AB - BACKGROUND: Adolescent health risk behaviors often occur together, suggesting that youth involvement with one risk behavior may inform understanding of other risk behaviors. We examined the association between involvement in nonsexual risk behaviors and trends among sexual behaviors. METHODS: We analyzed 1991-2007 data (n = approximately 125,000) from the Youth Risk Behavior Survey, a nationally representative survey of high school students in the United States. We categorized students into groups based on lifetime (Lifetime Risk Scale) and recent involvement (Recent Risk Scale) in nonsexual risk behaviors, such as smoking and drug use. We examined each group's prevalence of and trends for four sexual behaviors: ever having had sexual intercourse, having four or more lifetime partners, current sexual activity, and use of contraception at last sex. Data were examined for linear and quadratic (U-shaped) change using logistic regression. RESULTS: Sexual behaviors varied considerably between youth engaged in no risk behaviors and those in the highest risk behavior groups: sevenfold for ever having had intercourse (13% vs. 87% in 2007) and threefold for four or more lifetime sexual partners (19% vs. 57%). Despite these differences, trends in sexual risk behaviors among youth engaged in multiple nonsexual risk behaviors and those engaged in few or no risk behaviors were remarkably similar. In contrast, sexual behaviors demonstrated a very different pattern of change from that found or nonsexual behaviors: sexual experience and having multiple sexual partners declined into the early 2000s and then increased, whereas nonsexual behaviors increased over time, peaked in the late 1990 s, and then declined. CONCLUSIONS: Youth who engaged in little risk taking and those who engaged in considerable risk taking showed similar trends over time. However, the pattern of changes in sexual and nonsexual risk behaviors were remarkably different, raising questions about the potential impact of interventions that would reduce sexual risk taking by reducing nonsexual risk behaviors. Recent increases in sexual risk behaviors may have ominous implications for prevention of unplanned pregnancy and sexually transmitted infections among youth. PMID- 19306795 TI - Like parent, like child: intergenerational transmission of partner violence in Cebu, the Philippines. AB - PURPOSE: This study investigates the prevalence of partner violence perpetration and receipt among a sample of young men and women in the Philippines, as well as the relationship between witnessing interparental violence during childhood and current violence in partnerships. METHODS: We used 1994, 2002, and 2005 data from 472 married or cohabiting young adults from the Cebu Longitudinal Health and Nutrition Survey in Cebu, the Philippines. This is a longitudinal data set following more than 2000 Filipino women and their index children since the child's birth in 1983-1984. RESULTS: Prevalence of partner violence perpetration was 55.8% for female and 25.1% for male respondents. Prevalence of victimization was 27.7% for females and 30.5% for males. In all, 45% of females and 50% of males reported having witnessed their parents/caretakers physically hurt one another during childhood. Multinomial logistic regression analysis showed that witnessing interparental violence significantly predicted report of violent act victimization and reciprocal violent acts. Greater parental joint decision making and being male were independently associated with a lower risk of report of both reciprocal violent acts and violent act victimization. Duration of marriage or cohabitation was associated with report of violent act victimization and reciprocal violent acts. There were gender interaction effects for several factors, including mother's church attendance and household purchase of alcohol at age 11 years. CONCLUSIONS: Implications for further research and violence prevention programs include early intervention with adolescents and focus on gender differences in violence determinants. PMID- 19306797 TI - Spirituality and depressive symptoms in a school-based sample of adolescents: a longitudinal examination of mediated and moderated effects. AB - PURPOSE: To prospectively examine whether personal agency beliefs and direct coping mediate the association between spirituality and depressive symptoms in a school-based sample of adolescents, and whether gender, race, or grade level moderate this model. METHOD: Students (N = 1096) from sixth through ninth grades in a northeastern public school system were administered self-report instruments in group format at baseline, 6-month follow-up, and 1-year follow-up. Demographic variables and constructs of spirituality, personal agency, direct coping, and depressive symptoms were assessed. RESULTS: Structural equation modeling was used to examine the longitudinal associations among the constructs while controlling for socioeconomic status and baseline depressive symptoms. For the total sample, the model predicted 21% of the variance in depressive symptoms over 1 year. The model was moderated by gender but not by race or grade level. The model explained 28% of the variance in depressive symptoms for girls and 16% of the variance in depressive symptoms for boys. Moreover, there was an indirect effect of spirituality on depressive symptoms for girls but not for boys. CONCLUSION: These results suggest mechanisms by which spirituality may maintain lower levels of depressive symptoms among adolescent girls during periods of transition to middle and high school. PMID- 19306798 TI - Adolescent Tdap vaccine use among primary care physicians. AB - PURPOSE: In 2006 the Advisory Committee on Immunization Practices (ACIP) recommended replacement of the adolescent tetanus and diphtheria toxoids (Td) booster with combined tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis vaccine (Tdap). We examined the degree to which pediatricians and family practitioners have adopted this recommendation. METHODS: National mail based survey of a random sample of 725 pediatricians and 725 family practitioners from January through March, 2007. RESULTS: Overall response rate was 60%. The majority of respondents indicated they routinely recommended Tdap to adolescents at the preferred age for vaccination, 11-12 years old (87%), and also for "catch up" vaccination among adolescents 13-18 years old (89%). In bivariate analyses, pediatrician specialty, specialty society membership, stocking Tdap in the office, and prior experience diagnosing adolescent pertussis were associated with routinely recommending Tdap to adolescents. In multivariable models adjusting for these factors simultaneously, only pediatrician specialty (OR = 4.8, 95% CI = 2.5 9.3) and stocking Tdap in the office (OR = 14.5, 95% CI = 7.5-28.5) remained significantly associated with routine recommendation. Pediatricians were significantly more likely than family practitioners to accept shorter time intervals for administering Tdap following Td vaccination, and to co-administer Tdap with MCV4. Lack of adolescent visits was the most commonly cited major barrier to adolescent Tdap administration. CONCLUSIONS: Based on self report, our results indicate the majority of physicians have adopted recent recommendations from the ACIP to administer Tdap to adolescents. However, specialty-based disparities in attitudes and practices persist, suggesting that ongoing efforts are needed to motivate physicians to recommend this vaccine to adolescents and to clarify how to integrate Tdap with other adolescent vaccinations. PMID- 19306799 TI - Fertility preservation and adolescent/young adult cancer patients: physician communication challenges. AB - PURPOSE: The doctor-parent-adolescent triad is a unique communication challenge, particularly in the area of fertility preservation for adolescents with cancer. This paper provides a preliminary exploration into the barriers experienced by physicians in discussing cancer related fertility issues with patients aged 12 18. METHODS: This study used a subset of the data from qualitative interviews with pediatric oncologists. RESULTS: The majority of physicians agreed that fertility preservation conversations were awkward because of limited options and resources for the technology as well as the existence of a fine line between establishing a sense of trust between doctor and patient, while not excluding parents. CONCLUSIONS: Healthcare providers need training on how and when to broach fertility issues with patients, emphasizing open communication and early disclosure. PMID- 19306800 TI - Interactive roles of pubertal timing and peer relations in predicting social anxiety symptoms among youth. AB - The interaction between early maturation and problematic peer relations in relation to social anxiety symptoms was examined among 167 adolescents aged 10-17 years. Results indicated that early-maturing youth with problematic peer relations evidenced elevated social anxiety symptoms. Findings are discussed in terms of theoretical implications for adolescent social anxiety development. PMID- 19306801 TI - Attitudes and intentions to performing testicular self-examination: utilizing an extended theory of planned behavior. AB - This study examined the ability of an extended theory of planned behavior (TPB) to predict 500, 17- to 35-year-old male students' intentions to perform testicular self-examination (TSE). In a hierarchical regression analysis TPB components attitude, subjective norm and self-efficacy and the added constructs, past behavior, and anticipated regret all emerged as significant independent predictors of TSE intention. Overall, 64% of the variance in TSE intention was explained. Although prospective research is needed to confirm these preliminary findings, they suggest that an extended TPB might be a useful framework to guide interventions aimed at promoting TSE. PMID- 19306802 TI - Nonmedical use of prescription medications: an emerging risk behavior among rural adolescents. AB - Little is known about prescription medication abuse by rural youth. We surveyed 849 rural high school students and found that 34% reported lifetime nonmedical use of prescription medication higher than the 12% reported nationally. Boys and marijuana users had higher odds of nonmedical use of prescription medication than did girls or marijuana nonusers (odds ratio 1.9 and 3.8, respectively). PMID- 19306803 TI - Parenteral nutrition and anorexia nervosa: is it useful, is it ethical? PMID- 19306805 TI - Biliary tract cancers. Foreword. PMID- 19306806 TI - Biliary tract cancers. Preface. PMID- 19306807 TI - Malignant masquerade: dilemmas in diagnosing biliary obstruction. AB - The hepatobiliary surgeon must be as familiar with the nonmalignant processes that can affect the extrahepatic biliary tree as they are with the malignant causes. Subtleties in the patient's history, presentation, and imaging studies may prevent unnecessary extensive hepatobiliary resection. The focus of this article deals with the etiology of nonmalignant obstruction at the biliary bifurcation and hilum and the mid-bile duct. It does not focus on either choledocholithiasis or pancreatitis, the two most common causes of distal bile duct obstruction. Obstruction from pancreatic cancer is also not the focus of this discussion. PMID- 19306808 TI - Molecular mechanisms of cholangiocarcinogenesis: are biliary intraepithelial neoplasia and intraductal papillary neoplasms of the bile duct precursors to cholangiocarcinoma? AB - Cholangiocarcinoma (CC) is a rare, malignant neoplasm that can develop from any site within the intrahepatic or extrahepatic biliary tree. Although the key steps of cholangiocarcinogenesis remain unknown, it has been hypothesized that CC may develop through two key premalignant precursor lesions: biliary intraepithelial neoplasia (BilIN) and intraductal papillary neoplasm of the bile duct (IPNB). These lesions probably are analogous to pancreatic intraepithelial neoplasia and intraductal papillary mucinous neoplasm, respectively. This article outlines the molecular basis of cholangiocarcinogenesis through the BilIN and IPNB pathways. It highlights the genetic mutations that alter cellular proliferation, tumor suppression, and impairment of critical mucinous, cell-adhesion, and matrix proteins. PMID- 19306809 TI - Multimodality imaging of biliary malignancies. AB - Detection, accurate staging, and optimal management of biliary malignancies continue to present a significant challenge. This article reviews the current capabilities and roles of the various imaging modalities available in clinical practice, followed by a discussion of their integrated use at initial presentation, particularly with respect to potential surgical management of central hilar and intrahepatic cholangiocarcinoma. The main imaging modalities include MRI, CT, ultrasound, positron emission tomography, and conventional cholangiography. Alternative and emerging imaging methods, problematic diagnostic imaging issues, and other rarer bile duct malignancies are also briefly discussed. PMID- 19306810 TI - Percutaneous approach to the diagnosis and treatment of biliary tract malignancies. AB - The role of percutaneous, transhepatic management of biliary tract malignancies is to provide diagnostic and palliative care for improving patient quality of life. To treat and manage biliary tract malignancies successfully, particularly in patients who have inoperable disease, percutaneous interventions, such as biliary decompression and catheter-directed liver therapies, must be available. Although most vascular interventions do not yet commonly produce significant increased survivability in patients who have biliary tract malignancies, continued advances in percutaneous technologies suggest that future benefits in life expectancy may be achieved. PMID- 19306811 TI - Portal vein embolization in hilar cholangiocarcinoma. AB - In patients with hilar cholangiocarcinoma, extended hepatectomy and caudate lobe resection are often performed to achieve an R0 resection. In patients whose standardized future liver remnant is less than or equal to 20% of total liver volume, portal vein embolization (PVE) should be performed. In patients with biliary dilatation of the future liver remnant, a biliary drainage catheter should be placed before PVE. If the planned surgery is an extended right hepatectomy, segment 4 branch embolization improves the hypertrophy of segments 2 and 3. In high-volume centers, PVE can be safely performed; it increases the resectability rate and results in the same survival rates as those in patients who undergo resection without PVE. PMID- 19306812 TI - Extrahepatic cholangiocarcinoma: current surgical strategy. AB - Cholangiocarcinoma is a rare cancer. Although rare, it remains the second most common hepatobiliary cancer and its incidence is increasing worldwide. Extrahepatic cholangiocarcinoma can occur anywhere along the biliary tree and prognosis varies according to the location of disease. PMID- 19306813 TI - Management and extent of resection for intrahepatic cholangiocarcinoma. AB - Intrahepatic cholangiocarcinoma is the second most common primary liver cancer behind hepatocellular cancer. Although it is an uncommon malignancy, several reports have documented a significant rise in incidence, not only in the United States, but worldwide over the last several decades. As a result, interest in understanding the presentation, diagnosis, natural history, and efficacy of various treatment modalities has increased. This article reviews the clinical presentation, preoperative work-up, surgical management, and outcomes of patients undergoing resection for intrahepatic cholangiocarcinoma. Treatment options in unresectable patients are also reviewed. PMID- 19306814 TI - Surgical management of gallbladder cancer. AB - This article describes the epidemiology, risk factors, diagnostic imaging tools, and operative management of gallbladder cancer. The rarity of gallbladder cancer coupled with the prevalence of benign gallbladder disease mean that most patients undergo initial procedures that violate tumor planes, complicating attempts at future oncologic resection. Fortunately, a previous laparoscopic or open cholecystectomy does not lessen survival after definitive surgical extirpation. Large retrospective and underpowered prospective studies have suggested benefit to adjuvant chemotherapy or radiotherapy; however, these results need to be confirmed with large prospective randomized trials. PMID- 19306815 TI - Transplantation for cholangiocarcinoma: when and for whom? AB - Liver transplantation for cholangiocarcinoma has historically been maligned. Because of a high recurrence rate and poor patient survival, the disease has been viewed as an absolute contraindication to transplantation. Based on good results using neoadjuvant and palliative radiation, a protocol for liver transplantation in selected patients with unresectable hilar cholangiocarcinoma was developed in 1993. Neoadjuvant radiation is followed by operative staging to rule out patients with lymph node metastases before liver transplantation. This approach has achieved results superior to standard surgical therapy, with 72% 5-year survival for patients with unresectable disease. PMID- 19306816 TI - Radical resection of biliary tract cancers and the role of extended lymphadenectomy. AB - Extended hemihepatectomy and/or pancreatoduodenectomy plus extrahepatic bile duct resection and an extended lymphadenectomy of up to the group 2 lymph nodes can enable long-term survival in patients with extrahepatic bile duct (EBD) cancer with acceptable surgical risks. Surgeons should dissect and examine at least 10 or more nodes in curative intent surgeries for local disease control and accurate staging. Radical surgical procedures for EBD cancer, including a right lobectomy, left trisectoriectomy, hepatopancreatoduodenectomy, and combined vascular resection and reconstruction, are useful options for obtaining a negative margin, but the benefits of such procedures to long-term survival rates is limited to selected patients without nodal metastasis and with negative surgical margins. PMID- 19306817 TI - Indications for neoadjuvant, adjuvant, and palliative chemotherapy in the treatment of biliary tract cancers. AB - Advanced biliary tract carcinomas represent a group of aggressive diseases that still carries a poor prognosis. Chemotherapy has been shown to provide disease control and may also prolong survival. An established role for systemic therapy in the adjuvant setting is still lacking. This article reviews the available evidence to support indications of systemic chemotherapy in the palliative setting and discuss the attempts to study it in the perioperative settings. PMID- 19306818 TI - [Diabetes--Strasbourg 2009. Abstracts of Alfediam and Alfediam Paramedical meeting, 17-20 March, 2009, Strasbourg, France]. PMID- 19306822 TI - Two-year changes in health-related quality of life in gastric bypass patients compared with severely obese controls. AB - BACKGROUND: Few weight loss surgery trials have evaluated the changes in health related quality of life (HRQOL) relative to obese individuals not participating in weight loss interventions. In a prospective study at a bariatric surgery practice, we evaluated the 2-year changes in HRQOL in gastric bypass patients compared with 2 severely obese groups who did not undergo surgical weight loss. METHODS: A total of 308 gastric bypass patients were compared with 253 individuals who sought but did not undergo gastric bypass and 272 population based obese individuals using the weight-related (Impact of Weight on Quality of Life-Lite) and general (Medical Outcomes Study 36-item Short-Form Health Survey) HRQOL questionnaires at baseline and 2 years of follow-up. RESULTS: The percentage of weight loss was 34.2% for the gastric bypass and 1.4% for the no gastric bypass groups, with a .5% gain for population-based obese group. Both measures of HRQOL showed greater improvements for the gastric bypass group, even after controlling for baseline differences. Effect sizes for changes in physical and weight-related HRQOL were very large for gastric bypass, but small to medium for the 2 comparison groups. Effect sizes for changes in the psychosocial aspects of HRQOL were moderate to very large for gastric bypass, but small for the 2 comparison groups. Of the gastric bypass patients, 97% had meaningful improvements in the Impact of Weight on Quality of Life-Lite total score compared with 43% of the no gastric bypass group and 30% of the population-based obese group. CONCLUSION: Dramatic improvements had occurred in weight-related and physical HRQOL for gastric bypass patients at 2 years after surgery compared with 2 severely obese groups who had not undergone surgery. These results support the effectiveness of gastric bypass surgery in improving patients' HRQOL. PMID- 19306823 TI - Transoral endoscopic closure of gastric fistula. PMID- 19306824 TI - Comment on: Bile reflux after Roux-en-Y gastric bypass: an unrecognized cause of postoperative pain (Swartz DE, et al. 2009;5:27-30). PMID- 19306827 TI - Why not become an M.D.? A perspective from a student nurse on the profession's image. PMID- 19306828 TI - The ABCs of the doctor of nursing practice: assessing resources, building a culture of clinical scholarship, curricular models. AB - The Doctor of Nursing Practice (DNP) degree prepares nurses to provide comprehensive care across sites and over time. It is absolutely crucial-for both patient care and the nursing profession-that broadly recognized standards of competency for these new practitioners be established. The Council for the Advancement of Comprehensive Care has met since 2000 to build consensus on competency standards and a process for certifying these graduates. Deans of five nursing schools discuss their experiences and provide guidance for schools interested in developing DNP programs. PMID- 19306829 TI - Doctorate in nursing practice: a survey of massachusetts nurses. AB - Recently, the American Association of Colleges of Nursing (AACN) resolved that a new practice degree, the doctorate in nursing practice (DNP), is to become the terminal practice degree and minimum education standard for advanced practice nurses by the year 2015(American Association of Colleges of Nursing. (2004). AACN position statement on the practice doctorate in nursing. Retrieved July 1, 2007, from http://www.aacn.nche.edu.html). The DNP will have a clinical-intensive focus. Advanced practice nurses potentially impacted by this resolution will include nurse practitioners, nurse anesthetists, nurse midwives, and clinical nurse specialists. A task force at the William F. Connell School of Nursing at Boston College conducted an electronic survey in 2006 in an attempt to understand nurses' thoughts about doctoral preparation and the interest of nurses in Massachusetts in pursuing doctoral study. A self-selected group of 376 nurses participated in the study. Nurses identified both positive and negative perceptions related to the degree's viability and practicality, with a majority (55%) preferring the DNP as an educational option. PMID- 19306830 TI - Perceived stress and sense of belonging in doctor of nursing practice students. AB - There is little research among doctor of nursing practice (DNP) students, a fast growing population of nurses engaged in doctoral study. In this descriptive correlational study, levels of perceived stress, sense of belonging, and the relationship between these variables were examined. The sample included 89 female, predominantly White, post-master's DNP students from a Midwest university. A statistically significant inverse relationship (r = -.49, P < .01) between perceived stress and sense of belonging was found. Recommendations for future research include additional studies of perceived stress and sense of belonging in diverse DNP student populations and in various DNP education models. PMID- 19306831 TI - Strategies for successful management and oversight of nurse faculty workforce initiatives: lessons from the field. AB - A looming shortage of nursing faculty is a major contributor to the growing national and international nursing workforce shortage. The American Association of Colleges of Nursing (AACN, 2006, October) highlighted strategies in use nationally to address the shortage of nursing faculty in a recently posted Web site. Summarized on that site are responses from state-level nursing program administrators to a questionnaire generated by the AACN Government Affairs Committee. The questionnaire addressed the state-level management of nursing faculty/workforce initiatives including loan repayment options, scholarship rebates, in-state tuition benefits for out-of-state students, stipends, housing loans, and tax credits, intended to support nurse faculty recruitment and retention. Hermeneutic analysis of state-level descriptions suggested a set of strategies broadly representing ways to optimize success in managing faculty recruitment and retention. Specifically, the strategies were (a) interorganizational collaboration, (b) recognizing and leveraging local networks, (c) aligning stakeholder priorities, (d) sidestepping barriers, (e) thinking big, and (f) refusing to give up. This report describes the characteristics of these thematic strategies. PMID- 19306832 TI - Pros and woes of interdisciplinary collaboration with a national clinical trial. AB - Nursing collaborations with clinical trials provide opportunities to answer important nursing questions, enhance interdisciplinary collaboration, use collected data efficiently, include sophisticated measures, and answer questions that could not be answered by either project alone. To foster interdisciplinary collaboration, the National Institute of Nursing Research (NINR) issued a funding initiative to link nurse researchers and clinical trial teams. This article describes a successful collaboration between Extremity Constraint-Induced Therapy Evaluation (EXCITE), a multisite, national clinical trial, and Caregiver-EXCITE, a complementary nursing study funded by NINR. EXCITE tested a neurorehabilitative technique to improve upper extremity function in stroke survivors, whereas Caregiver-EXCITE addressed hypotheses about family function, stroke recovery, and caregiver outcomes. This article addresses advantages and challenges of this collaboration and provides illustrations of excellent opportunities that joint ventures offer to advance science. These types of collaborations have the potential to profoundly move nursing's research agenda forward. PMID- 19306833 TI - Peer reviewer training and editor support: results from an international survey of nursing peer reviewers. AB - TOPIC: Nursing journals depend on the services of peer reviewers for their expertise in research and clinical practice. Although some research has been done with peer reviewers of biomedical journals, to date, our knowledge about reviewers of nursing journals is minimal. METHODS: In this international survey of 1,675 reviewers for 41 nursing journals, reviewers were asked 69 questions about their experiences reviewing for professional nursing journals. This article examines their answers to the survey questions about training to become reviewers and the support they receive from editors. RESULTS: Results showed that 65% wanted formal training, although only about 30% received such training in the form of orientation, manuals, practice reviews, or workshops. For most peer reviewers, it took one to five reviews before they felt comfortable with the process, although some commented that, "I still question my reviews" and "It took a few years." In this sample, 31% reported getting feedback from editors about their reviews, but 87% wanted feedback. Most (80%) wanted to see the other reviews of the manuscripts they reviewed, although only about 45% actually saw them. Reviewers reported that the editor had been helpful to them by providing feedback, demonstrating appreciation of their efforts, mentoring, and being available. CONCLUSIONS: We concluded from this research that many reviewers' needs for training and support are not being met and that both reviewers and nursing editors could profit from a better understanding of the process. Editors could consider instituting programs of orientation, training, and support such as feedback on reviews, making other reviews available, and feedback on final disposition of manuscripts. Reviewers should consider discussing these issues with editors to make their needs for feedback and training known. Intervention studies to examine the effects of such programs on reviewer satisfaction could ultimately strengthen the nursing literature. PMID- 19306834 TI - An investigation of nursing competence and the competency outcomes performance assessment curricular approach: senior students' self-reported perceptions. AB - This study was designed to measure the construct of competence as perceived by nursing students in different programs of study. An exploration of a competence based curricular model was undertaken to determine if there are distinguishing differences between schools that have formally adopted Dr. Carrie Lenburg's Competency Outcomes Performance Assessment (COPA) model as a curricular guide and those who have do not have a formal competence model guiding the curriculum. The Six-Dimension Scale of Nursing Performance (6-D Scale) was used to evaluate nursing competence. Using a purposive sample, senior nursing students (N = 391) from three different types of nursing programs (baccalaureate, associate [ADN], and diploma) were surveyed. Students completed a two-part questionnaire that included the 6-D Scale, demographic questions, and a researcher-generated questionnaire designed to measure competency-based learning and evaluation. Faculty (n = 101) from these same programs were also surveyed. The results revealed that students from COPA and non-COPA schools reported slightly lower scores in three subscales: teaching/collaboration, critical care, and leadership. Significant curricular differences as defined by the Klein Scales were found between COPA and non-COPA senior students, with 17% to 30% of the variance explained for the COPA students. The findings reflect that baccalaureate students reported significantly lower 6-D Scale scores in multiple areas when compared to diploma and ADN students. Enhancement of student learning and curricular revisions designed to improve leadership and technical expertise are suggested by the findings of this study. PMID- 19306835 TI - Nurses of African descent and career advancement. AB - The purpose of this article is to evaluate a leadership institute designed to promote career advancement and leadership in administration, education, and research among nurses of African descent. Government reports indicate that Black Americans receive lower quality health care than other racial groups even when insurance and income are equal. Moreover, the literature suggests that less than 10% of practicing professional nurses in America are Black-and of these, less than 1% are in senior executive leadership positions. However, the literature lacks detailed discussion of the effectiveness of leadership programs. This article provides an in-depth look at a leadership institute for Black nurses and outlines the impact of the program. PMID- 19306836 TI - A nonradioactive dot blot assay for transglutaminase activity. AB - Aberrant transglutaminase (TG) activity has been implicated in the pathology of numerous diseases, including Huntington's disease and Alzheimer's disease. To fully characterize the role of TGs in these disorders, it is important that simple quantifiable assays be made available. The most commonly used assay currently employed requires significant time and a radioactive substrate. The assay described here uses a biotinylated substrate in conjunction with a dot blot apparatus to eliminate the use of radioactive substrates and allows relative transglutaminase activity to be measured simultaneously with minimal sample preparation in a large number of samples containing purified enzyme, cell extracts, or tissue homogenates. PMID- 19306837 TI - Cobalt hydroxide nanoparticles modified glassy carbon electrode as a biosensor for electrooxidation and determination of some amino acids. AB - The electrochemical behavior of some amino acids was investigated on cobalt hydroxide nanoparticles modified glassy carbon (CHM-GC) electrode in alkaline solution. The process of oxidation and its kinetics were established by using cyclic voltammetry, chronoamperometry techniques, and steady-state polarization measurements. The results revealed that cobalt hydroxide promotes the rate of oxidation by increasing the peak current, so these bimolecular reactions are oxidized at lower potentials. Cyclic voltammograms and chronoamperometry indicate a catalytic EC' mechanism to be operative with electrogeneration of Co(IV) as the electrochemical process. Also, the process is diffusion controlled and the current-time responses follow Cottrellian behavior. This result was confirmed by steady-state measurements. The rate constants of the catalytic oxidation of amino acids and the electron transfer coefficients are reported. PMID- 19306838 TI - Correlations between in vitro potency of polyethylene glycol-protein conjugates and their chromatographic behavior. AB - Pegylation is the most widely used and accepted methodology for half-life extension of biopharmaceutical drugs that also improves physicochemical and biological characteristics of proteins considerably. Most of the positive pharmacological effects of pegylated proteins are believed to be related to an increased hydrodynamic volume and molecular size. To explore the size impact of polyethylene glycol (PEG) on in vitro potency, a series of well-defined conjugates of interferon alpha-2b (IFN) were prepared with PEGs of different lengths and shapes specifically attached to the N-terminal amino group of the protein. Specificity of the attachment was confirmed by peptide mapping and mass spectroscopy. When potency values determined by reporter gene assay were correlated with methods for molecular weight and size characterization, such as size exclusion chromatography and dynamic light scattering, rough parallels were found. Unexpectedly, the retention times on cation exchange chromatography showed much higher correlation with experimentally determined in vitro potency. It appears that in a series of N-terminally pegylated IFNs, their in vitro potency could be predicted from the retention times on the cation exchange chromatography columns, probably because both methods reflect not only the influence of molecular size but also the impact of protein masking exerted by attached PEG moiety. PMID- 19306839 TI - Novel copper amine oxidase activity from rat liver mitochondria matrix. AB - Copper containing amine oxidases (Cu-AO) represent a heterogeneous class of enzymes classified as EC 1.4.3.6. The present study reports preliminary results on the presence of a novel amine oxidase activity in rat liver mitochondria lysates. Such enzymatic activity was found in the soluble mitochondrial fraction, obtained by simple osmotic shock. The mitochondrial amine oxidase was isolated by affinity chromatography on a newly synthesised spermine-Sepharose. SDS-PAGE showed a single band at about 60kDa. Upon chromatographic purification, the enzymatic activity was very labile. The crude enzyme activity was tested by spectrophotometric measurements, determining hydrogen peroxide production following oxidative deamination of different substrates, such as polyamines (spermine, spermidine, putrescine and cadaverine) and monoamines (dopamine and benzylamine). The activity, observed on polyamines and not on monoamines, was inhibited by semicarbazide and azide, but not by pargyline, clorgyline and l deprenil. Enzyme specificity was tested on several diamines characterized by different carbon atom chain length in the range 2-6 carbon atoms. The highest activity was found with 1,2-diamino-ethane and the highest affinity with 1,5 diamino-pentane. The above reported results suggest the presence of a novel copper-dependent amine oxidase in liver mitochondria matrix. PMID- 19306840 TI - The local electric field within phospholipid membranes modulates the charge transfer reactions in reaction centres. AB - Three different cholesterol derivatives and phloretin, known to affect the local electric field in phospholipid membranes, have been introduced into Rhodobacter sphaeroides reaction centre-containing phospholipid liposomes. We show that cholesterol and 6-ketocholestanol significantly slow down the interquinone first electron transfer (approximately 10 times), whereas phloretin and 5-cholesten 3beta-ol-7-one leave the kinetics essentially unchanged. Interestingly, the two former compounds have been shown to increase the dipole potential, whereas the two latter decrease it. We also measured in isolated RCs the rates of the electron and proton transfers at the first flash. Over the pH range 7-10.5 both reactions display biphasic behaviors with nearly superimposable rates and amplitudes, suggesting that the gating process limiting the first electron transfer is indeed the coupled proton entry. We therefore interpret the effects of cholesterol and 6-ketocholestanol as due to dipole concentration producing an increased free energy barrier for protons to enter the protein perpendicular to the membrane. We also report for the first time in R. sphaeroides RCs, at room temperature, a biphasicity of the P(+)Q(A)(-) charge recombination, induced by the presence of cholesterol derivatives in proteoliposomes. We propose that these molecules decrease the equilibration time between two RC conformations, therefore revealing their presence. PMID- 19306841 TI - Expression of purinergic receptors and modulation of P2X7 function by the inflammatory cytokine IFNgamma in human epithelial cells. AB - The cervical epithelial cell line, HeLa, is one of the oldest and most commonly used cell lines in cell biology laboratories. Although a truncated P2X(7) receptor has recently been identified in HeLa cells, the expression of other purinergic receptors or the function of the P2X(7) protein has not been characterized. We here show that HeLa cells express transcripts for most P2X and P2Y purinergic receptors. Treatment of cells with ATP or other P2X(7) agonists does not stimulate cell death, but can induce atypical calcium fluxes and ion currents. Cervical epithelial cells represent an important target for sexually transmitted pathogens and are commonly exposed to pro-inflammatory cytokines such as IFNgamma. Stimulation of HeLa cells with IFNgamma upregulates expression of P2X(7) mRNA and full-length protein, modifies ATP-dependent calcium fluxes, and renders the cells sensitive to ATP-induced apoptosis, which can be blocked by a P2X(7) antagonist. IFNgamma treatment also increased dramatically the sensitivity of the intestinal epithelial cell line, HCT8, to ATP-induced apoptosis. Significantly, IFNgamma also stimulated P2X(7) expression on human intestinal tissues. Responses to other purinergic receptor ligands suggest that HeLa cells may also express functional P2Y(1), P2Y(2) and P2Y(6) receptors, which could be relevant for modulating ion homeostasis in the cells. PMID- 19306842 TI - Endothelial nitric oxide synthase activation contributes to post-exercise hypotension in spontaneously hypertensive rats. AB - We investigated the role that endothelial nitric oxide synthase plays in post exercise hypotension in spontaneously hypertensive rats. To accomplish this, rats were subjected to a single bout of dynamic exercise on a treadmill at 15 m/min for 20 min. L-nitroarginine methyl ester (L-NAME, 40 mg/kg, i.p.) significantly inhibited post-exercise hypotension (25+/-11 and 5+/-3 mm Hg, respectively; P<0.05). In addition, the superoxide anion generation was decreased, while the plasma nitrite production and serine phosphorylation of endothelial nitric oxide synthase were significantly elevated in spontaneously hypertensive rats at 30 min after the termination of exercise. Taken together, these data demonstrate that the increased phosphorylation of endothelial nitric oxide synthase plays a crucial role in the reduction of arterial pressure following a single bout of dynamic exercise in spontaneously hypertensive rats. PMID- 19306843 TI - Knock-out of Arabidopsis AtNHX4 gene enhances tolerance to salt stress. AB - AtNHX4 belongs to the monovalent cation:proton antiporter-1 (CPA1) family in Arabidopsis. Several members of this family have been shown to be critical for plant responses to abiotic stress, but little is known on the biological functions of AtNHX4. Here, we provide the evidence that AtNHX4 plays important roles in Arabidopsis responses to salt stress. Expression of AtNHX4 was responsive to salt stress and abscisic acid. Experiments with CFP-AtNHX4 fusion protein indicated that AtNHX4 is vacuolar localized. The nhx4 mutant showed enhanced tolerance to salt stress, and lower Na(+) content under high NaCl stress compared with wild-type plants. Furthermore, heterologous expression of AtNHX4 in Escherichia coli BL21 rendered the transformants hypersensitive to NaCl. Deletion of the hydrophilic C-terminus of AtNHX4 dramatically increased the hypersensitivity of transformants, indicating that AtNHX4 may function in Na(+) homeostasis in plant cell, and its C-terminus plays a role in regulating the AtNHX4 activity. PMID- 19306844 TI - Fluorescence in situ hybridization and chromosomal organization of the sirtuin 4 gene (Sirt4) in the mouse. AB - The sirtuins (SIRT1-7), also being referred to as class III HDACs, exert NAD dependent deacetylase and/or ADP-ribosyltransferase activities in various cellular compartments including the cell nucleus, the cytoplasm and the mitochondria. The sirtuins play a central role in epigenetic gene silencing, DNA repair and recombination, cell-cycle, microtubule organization, and in the regulation of aging. SIRT4 is a mitochondrial protein that lacks deacetylase activities but efficiently works as an ADP-ribosyltransferase. We have isolated and characterized the murine Sirt4 genomic sequence, which spans a region of 12kb and which has one single genomic locus. Determination of the exon-intron splice junctions established that SIRT4 is encoded by 6 exons. The 1648bp murine Sirt4 transcript encodes a 418 aa protein with a predictive molecular weight of 47.3kDa. Fluorescence in situ hybridization analysis identified a single genomic locus for murine Sirt4 gene on chromosome 5F and is neighbored by the PLA2G1B and PXN genes. PMID- 19306845 TI - Fluorescence-based optimization of human bitter taste receptor expression in Saccharomyces cerevisiae. AB - Human TAS2 receptors (hTAS2Rs) perceive bitter tastants, but few studies have explored the structure-function relationships of these receptors. In this paper, we report our trials on the large-scale preparations of hTAS2Rs for structural analysis. Twenty-five hTAS2Rs were expressed using a GFP-fusion yeast system in which the constructs and the culture conditions (e.g., the signal sequence, incubation time and temperature after induction) were optimized by measuring GFP fluorescence. After optimization, five hTAS2Rs (hTAS2R7, hTAS2R8, hTAS2R16, hTAS2R41, and hTAS2R48) were expressed at levels greater than 1mg protein/L of culture, which is a preferable level for purification and crystallization. Among these five bitter taste receptors, hTAS2R41 exhibited the highest detergent solubilization efficiency of 87.1% in n-dodecyl-beta-d-maltopyranoside (DDM)/cholesteryl hemisuccinate (CHS). Fluorescence size-exclusion chromatography showed that hTAS2R41 exhibited monodispersity in DDM/CHS without aggregates, suggesting that hTAS2R41 is a good target for future crystallization trials. PMID- 19306846 TI - Is conceptual processing in music automatic? An electrophysiological approach. AB - Recent evidence suggests that music perception, much alike language perception, involves the cognitive processing of concepts, that is abstract general ideas. In a previous study (Daltrozzo and Schon, Conceptual processing in music as revealed by N400 effects on words and musical targets. Journal of Cognitive Neuroscience, In Press), we reported the effect of the presentation of a musical excerpt (the context) on the perception of a word, while participants judged the conceptual relatedness between the two stimuli. Event-Related Potentials (ERPs) showed a N400 effect: a larger N400 to words judged unrelated to their context compared to related words. In the present experiment, we decided to test the influence of the relatedness task on the N400 effect by using a more implicit task: lexical decision. We recorded behavioral and ERP data while participants were presented 50 related and 50 unrelated pairs (excerpt context/word target). An N400 effect was again observed. However, the N400 effect found with a lexical decision was more than two times smaller than with a relatedness judgment task and was significant in a later latency range: 500 to 650 ms instead of 300 to 550 ms with a relatedness judgment. These differences are interpreted as reflecting the task induced modulation of explicit (strategic) mechanisms involved in the N400 effect. PMID- 19306847 TI - Region-specific susceptibilities to cuprizone-induced lesions in the mouse forebrain: Implications for the pathophysiology of schizophrenia. AB - Cuprizone (CPZ) is a neurotoxic agent acting as a copper chelator. In our recent study, C57BL/6 mice given dietary CPZ (0.2%) showed impairments in spatial working memory, social interaction, and prepulse inhibition. These abnormalities are reminiscent of certain schizophrenia symptoms and are not likely due to damage in the whole brain or in any single white matter tract/brain region. We hypothesized that white matter damage resulting from CPZ-treatment may be site specific rather than universal. We examined the forebrains of C57BL/6 mice given the CPZ-containing diet and compared them with those of controls. We assessed CPZ induced demyelination in main white matter tracts of the forebrain, evaluated myelin break down in the neuropil of the main olfactory bulb (MOB), cerebral cortex (CTX), caudate putamen (CP), hippocampus (HP), thalamus (TH), and hypothalamus (HY), and counted the number of myelin sheath forming oligodendrocytes (OLs) in CTX, CP, TH, and HY. Obvious demyelination was observed in the corpus callosum, external capsule, CP, and dorsal hippocampal commissure whereas other tracts seemed to be unaffected. The neuropil of CTX, HP and MOB showed myelin break down, which was mild in TH and HY. The number of OLs was decreased in all the above regions of CPZ-treated mice although the degree of OL loss was not consistent across regions. The data provide further support for white matter abnormalities contributing to schizophrenia-like behaviors in mice. PMID- 19306848 TI - Taxonomic and thematic categories: Neural correlates of categorization in an auditory-to-visual priming task using fMRI. AB - Categorization is a basic principle of knowledge organization in the brain. The goal of the current fMRI study was to compare the neural correlates of thematic (e.g., car - garage) and taxonomic (e.g., couch - bed) categories under automatic processing conditions using auditory-to-visual semantic priming. Behavioral data revealed a priming effect for thematically but not for taxonomically related word pairs. On a neural level, thematically related words led to a left-lateralized temporal activation (superior temporal sulcus), whereas taxonomically related word pairs evoked a right-lateralized frontal activation and within the hippocampus. A direct comparison between both categories revealed enhanced activation for thematically related and response suppression for taxonomically related trials in the left superior temporal sulcus. These results suggest that processing of thematic and taxonomic categories leads to activation of distinct brain areas. The mainly right-lateralized fronto-temporal activation for taxonomic relations suggests increased attention and effort for processing this category. The interaction within the left superior temporal sulcus reflects the processing and retrieval of semantic relations whereby specific memory contents seem to influence the direction of activation. PMID- 19306849 TI - Glibenclamide improves neurological function in neonatal hypoxia-ischemia in rats. AB - Recent studies demonstrated that sulfonylurea receptor 1 (SUR 1) regulated nonselective cation channel, the NC(Ca-ATP) channel, is involved in brain injury in rodent models of stroke. Block of SUR 1 with sulfonylurea such as glibenclamide has been shown to be highly effective in reducing cerebral edema, infarct volume and mortality in adult rat models of ischemic stroke. In this study, we tested glibenclamide in both severe and moderate models of neonatal hypoxia-ischemia (HI) in postnatal day 10 Sprague-Dawley rat pups. A total of 150 pups were used in the present study. Pups were subjected to unilateral carotid artery ligation followed by 2.5 or 2 h of hypoxia in the severe and moderate HI models, respectively. In the severe HI model, glibenclamide, administered immediately after HI and on postoperative Day 1, was not effective in attenuating short-term effects (brain edema and infarct volume) or long-term effects (brain weight and neurological function) of neonatal HI. In the moderate HI model, when injected immediately after HI and on postoperative Day 1, glibenclamide at 0.01 mg/kg improved several neurological parameters at 3 weeks after HI. We conclude that glibenclamide provided some long-term neuroprotective effect after neonatal HI. PMID- 19306850 TI - The influence of mental fatigue and motivation on neural network dynamics; an EEG coherence study. AB - The purpose of the present study is to examine the effects of mental fatigue and motivation on neural network dynamics activated during task switching. Mental fatigue was induced by 2 h of continuous performance; after which subjects were motivated by using social comparison and monetary reward as motivating factors to perform well for an additional 20 min. EEG coherence was used as a measure of synchronization of brain activity. Electrodes of interest were identified using a data-driven pre-processing method (ten Caat, M., Lorist, M.M., Bezdan, E., Roerdink, J.B.T.M., Maurits, N.M., 2008a. High-density EEG coherence analysis using functional units applied to mental fatigue. J. Neurosci. Meth. 171, 271 278; ten Caat, M., Maurits, N.M. and Roerdink, J.B.T.M., 2008b. Data-driven visualization and group analysis of multichannel EEG coherence with functional units. IEEE T. Vis. Comp. Gr. 14, 756-771). Performance on repetition trials was faster and more accurate than on switch trials. EEG data revealed more pronounced, frequency specific fronto-parietal network activation in switch trials, while power density was higher in repetition trials. The effects of mental fatigue on power and coherence were widespread, and not limited to specific frequency bands. Moreover, these effects were independent of specific task manipulations. This increase in neuronal activity and stronger synchronization between neural networks did not result in more efficient performance; response speed decreased and the number of errors increased in fatigued subjects. A modulation of the dopamine system is proposed as a common mechanism underlying the observed the fatigue effects. PMID- 19306851 TI - Numerical magnitude modulates temporal comparison: an ERP study. AB - Time is believed to be a part of the generalized magnitude system just like space and quantity. Previous research suggests that time perception can be affected by magnitude in some non-temporal dimensions. Here we address two questions. First, could the influence be caused by an abstract magnitude component without perceptual variables? Second, what are the underlying mechanisms of the influence? Participants compared a pair of durations defined by two Arabic digits in a hundreds of milliseconds range. They performed more accurately when the shorter durations were defined by lower numeric value digits (small digits) and the longer durations were defined by higher value digits (large digits) than they did in the reversed condition. Event-Related Potential (ERP) results showed that the CNVs corresponding to the first duration (CNV1), to the second duration (CNV2) and the N1 were all enhanced when durations marked by small digits than that marked by large ones. Combining the electrophysiological data with the behavioral results, we suggest that digits can modulate performance of temporal comparison at the relatively early stage of perceptual processing. One possible explanation of the current results is that selective temporal attention and subsequent expectation may be involved in this modulation. PMID- 19306852 TI - Nestin-positive microglia in adult rat cerebral cortex. AB - Nestin is a class VI intermediate filament protein, which was first identified in the early developmental stages of the nervous system. It is widely used as a stem or progenitor cell marker. In the adult mammalian brain, nestin is expressed not only in germinal cells in the neurogenic regions but also in non-germinal cells, such as reactive astrocytes, endothelial cells and pericytes. In the present study, we found another nestin-positive cell type within the adult rat cerebral cortex. We immunohistochemically analyzed which types of cells exhibit immunoreactivity for nestin, and through the use of co-immunostaining with Iba1, CD11b and GLUT5, which are known to be specific for microglia, identified these cells as microglia. Approximately >20% of the microglia were immunoreactive for nestin in the rat cerebral cortex under normal conditions. Nestin signals were not widely distributed in the microglial cytoplasm, but were restricted to the perikaryon and to parts of the cell processes. Nestin-positive microglia were also immunoreactive for the intermediate filament protein vimentin. These observations demonstrate that a subpopulation of microglia in a resting state has nestin-containing intermediate filament networks. Therefore, nestin in conjunction with vimentin might have roles in maintaining the structural integrity of the microglia. PMID- 19306853 TI - Long-term increasing co-localization of SCN8A and ankyrin-G in rat hippocampal cornu ammonis 1 after pilocarpine induced status epilepticus. AB - Voltage-gated sodium channels (VGSC) are important determinants of neuronal excitability which are implicated in the pathogenesis of epilepsy. Ankyrin-G contributes to the distribution and regulation of VGSC. Here we investigated the alterations of the two alpha-subunits SCN8A and SCN1A and their adapter ankyrin-G in the hippocampal cornu ammonis 1 (CA1) of rats after pilocarpine induced status epilepticus (PISE), compared to the sham-control group (C1) and blank-control group (C2). Significant increase of SCN8A mRNA (41.08% increase compared to C1, P<0.001; 30.88% increase compared to C2, P=0.011) was detected 60 days after PISE. At D1 SCN8A mRNA reduced but no significant changes were detected when compared to controls (one-way ANOVA, F=1.232, P=0.276). After measuring the optical density of Western blot, we detected significant differences between the levels of SCN8A protein in different groups but no difference between the protein levels of SCN1A at D1 and D60 after pilocarpine treatment compared to the control. At D60 the relative copies of ankyrin-G mRNA on internal control beta actin in PISE group increased significantly compared to C1 and C2 (one-way ANOVA, F=16.537, P<0.001). Significantly increase of ankyrin-G immunoreactivity in Western blot from the PISE group 1 day and 60 days after PISE was observed, compared to the controls (one-way ANOVA, F=24.255 at D1, P<0.001; F=29.280 at D60, P<0.001). After analyzing the double-stained cells counting, we detected significant differences between the numbers of SCN8A+/ankyrin-G+ immunoreactive cells in different groups in acute and chronic period following PISE (two way ANOVA, F(group)=37.905, P<0.001; F(day)=45.310, P<0.001). The data revealed that both SCN8A and ankyrin-G increased significantly in the CA1 subfield of the rat hippocampus 60 days following pilocarpine induced status epilepticus and co localized with each other. PMID- 19306854 TI - Interference control during recognition of facial affect enhances the processing of expression specific properties--an event-related fMRI study. AB - Though we can almost pre-attentively categorize the valence of facial expressions, we experience emotional ambiguity when confronted with facial expressions in a context with incongruent emotional information. We simultaneously presented interfering background colors during forced-choice categorizations of negative (fear), neutral and positive (happy) expressions. Conflicting information induced strong and differential interference effects on a behavioral level which was mirrored in comparable activations on a neuronal level. Besides a common fronto-parietal attention network which was activated during interference resolution, we found differential interference effects for facial expressions. Incongruent trials with neutral expressions induced a distinct activation pattern in ventral visual regions particularly involved in deeper analysis for both the task-relevant facial expressions (fusiform (FFA) and occipital face area (OFA)) and the task-irrelevant color (V4). Compared to neutral expressions, incongruent trials including either negative or positive expressions elicited attenuated interference effects. Unlike incongruent trials with positive facial expressions which showed only sparse activation in frontal cortex, interference resolution during processing of negative facial expressions resulted in specific activations in regions (V3a, MT(+), STS) which might be involved in processing of implicit dynamics of negative expressions. Thus, functional activations in visual processing regions might specifically be related to processing demands of different expressions. PMID- 19306855 TI - N250r ERP repetition effects from distractor faces when attending to another face under load: Evidence for a face attention resource. AB - Recently, evidence for a face-specific attentional resource was suggested, which limits simultaneous processing to only one face. In the present Experiment 1, we manipulated perceptual load using two central item types (CITs: small central buildings or unfamiliar faces). To test whether distractor face processing is effectively prevented by face targets, CITs were superimposed on large famous distractor faces. ERPs were measured to subsequent faces, which could be a repetition or non-repetition of the previous distractor face. In Experiment 2, we used famous and unfamiliar faces as CITs under high load. For building CITs, we found common N250r repetition effects both under high and low load. For face CITs, N250r was reduced (Experiment 1) or even eliminated (Experiment 2) under high load. These findings support notions of a face-specific attentional resource which, at least under high demands, may limit processing to only one face at a time. PMID- 19306856 TI - Performance characteristics of laboratory testing and clinical outcomes. AB - In order to demonstrate the relationship between performance characteristics of laboratory tests and clinical outcomes, diabetes seems to represent a paradigmatic disease: diagnosis, monitoring of therapeutic efficacy and prognosis are adequately achieved by means of laboratory testing. Starting from a simple molecule, glucose, used for the diagnosis of diabetes, continuing with creatinine, used for monitoring renal function in diabetic patients and concluding with cardiac troponins, a recognised gold standard for the diagnosis and risk stratification of cardiovascular diseases, several criticisms may be stressed considering the current methodological state-of-the art. Finally, an often overlooked aspect of performance, the analytical interferences, being responsible of unexpected results, that in turn depend from unknown or undisclosed factors will be discussed, concerning in particular, in our paper, the macroprolactin and the heterophilic antibodies aspects. PMID- 19306857 TI - Analytical interferences and analytical quality. AB - In today's health care system the prevalence of medical errors is high as stated by the report of the Institute of Medicine. A varying error rate of <10% in clinical medical laboratories has been reported in the literature. Most of these errors occur in the pre-analytical phase. Only a small number of errors will be seen in the analytical phase. This overview will deal with the analytical interferences and will offer ways to improve the analytical quality. Some special areas of the analytical process like calibration, quality control, reference interval, drug interference, statistical analysis and volume displacement will be covered. With some examples from the literature and own investigations the impact of errors in these steps of the analytical process will be better understood and the examples will help reducing the number of analytical errors and interferences. This finally provides better patient safety. PMID- 19306858 TI - A novel missense HGD gene mutation, K57N, in a patient with alkaptonuria. AB - Alkaptonuria is a rare recessive disorder of phenylalanine/tyrosine metabolism due to a defect in the enzyme homogentisate 1,2-dioxygenase (HGD) caused by mutations in the HGD gene. We report the case of a 38 year-old male with known alkaptonuria who was referred to an adult metabolic clinic after initially presenting to an emergency department with renal colic and subsequently passing black ureteric calculi. He complained of severe debilitating lower back pain, worsening over the last few years. A CT scan revealed marked degenerative changes and severe narrowing of the disc spaces along the entire lumbar spine. Sequencing of the HGD gene revealed that he was a compound heterozygote for a previously described missense mutation in exon 13 (G360R) and a novel missense mutation in exon 3 (K57N). Lys(57) is conserved among species and mutation of this residue is predicted to affect HGD protein function by interfering with substrate traffic at the active site. In summary, we describe an alkaptonuric patient and report a novel missense HGD mutation, K57N. PMID- 19306859 TI - Proteomics-based identification of alpha1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas. AB - BACKGROUND: The identification of pathological markers of breast cancer for either diagnosis, treatment response or for survival is of critical importance. METHODS: Serum protein profiling using 2-DE separations coupled to matrix assisted laser desorption ionization mass spectrometry has been used to explore protein alterations in patients with infiltrating ductal breast carcinomas (IDCA). Sera from 39 breast cancer patients and 40 healthy controls were selected for screening study using 2-DE combined with MS. The protein expression patterns obtained after the depletion of high abundance proteins was determined by coomassie blue G-250 stain after 2-DE electrophoresis. RESULTS: Six proteins that expressed differentially in the IDCA group were found. The expression levels of four isoforms corresponding to haptoglobin precursor and two isoforms of alpha1 antitrypsin precursor (alpha1-AT) were upregulated in sera from breast cancer patients. There was an increased expression of both proteins in the sera of patients with various tumor stages (I, II, III) in comparison to healthy women. Applying immunohistochemistry, we further validated alpha1-AT immunoreactivity in 51 formalin-fixed paraffin-embedded sections of breast tumors. Enhanced expression of alpha1-AT like activity has been found in IDCA breast tumors, as well as, in different histological types of breast cancer. No significant association has been found with lymph node occurrence, while in high tumor categories a tendency to an increased expression of alpha1-AT has been found, thereby suggesting a possible role of this protein in tumor growth. CONCLUSIONS: These proteins may constitute new and useful markers of breast cancer that offer a clue to a better understanding of inflammatory pathways and carcinogenesis events linked to breast cancer progression. PMID- 19306860 TI - Interpretative reports and critical values. AB - In the clinical laboratory to allow an effective testing process, post-analytical activity can have two goals in trying to improve patient safety: result interpretation and communication of critical values. Both are important issues, and their success requires a cooperative effort. Misinterpretation of laboratory test results or ineffectiveness in their notification can lead to diagnostic errors or errors in identifying patient critical conditions. With the awareness that the incorrect interpretation of tests and the breakdown in the communication of critical values are preventable errors, laboratorians should make every effort to prevent the types of errors that potentially harm patients. In order to improve the reliability of laboratories, we attempt to explain how interpretative reporting and automated notification of critical values can be used to reduce errors. Clinical laboratories can therefore work to improve clinical effectiveness, without forgetting that everything should be designed to provide the best outcomes for patients. PMID- 19306861 TI - Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum. AB - BACKGROUND: Serum deoxyribonuclease I (DNase I) activity was reported to increase in the early phase after onset of acute myocardial infarction (AMI). Up to now, DNase I activity has been quantified by the single radial enzyme diffusion (SRED) method, which unfortunately requires a long incubation time. Therefore it is necessary to develop another assay suitable for measurement of serum DNase I concentrations in a clinical setting. METHODS: A sandwich ELISA was established for measurement of DNase I protein using a polyclonal antibody directed against DNase I protein and a biotinylated monoclonal for subsequent detection. Concentrations of serum DNase I protein were measured in healthy individuals and patients with AMI. RESULTS: This method was as precise as SRED, and took less time than SRED. A significant correlation was observed between DNase I concentration and enzyme activity (r=0.839; P<0.001). The average of serum DNase I in AMI patients within 0-12 h of chest pain was significantly higher than that in healthy individuals (P<0.001), and decreased with time. CONCLUSIONS: We have developed a sensitive ELISA capable of measuring DNase I protein concentrations. This method may be a useful alternative to SRED as an aid to diagnosis of AMI based on the serum DNase I level. PMID- 19306862 TI - Metabolic biomarkers related to oxidative stress and antioxidant status in Saudi autistic children. AB - OBJECTIVE: Measurement of oxidative stress and antioxidant-related parameters (enzymatic and non-enzymatic) in Saudi autistic children. DESIGN AND METHODS: 30 autistic children (22 males and 8 females) aged 3-15 years (25/30 of these were below 8 years old), and 30 healthy children as control group were included in this study. Levels of lipid peroxides, vitamin E, vitamin C, glutathione together with enzymatic activities of glutathione peroxidase (GSH-Px), and catalase were determined in plasma while superoxide dismutase (SOD was measured in red blood cells of both groups. RESULTS: Lipid peroxidation was found to be significantly higher in autistic compared to control Saudi children. On the other hand, vitamin E and glutathione were remarkably lower in autistic patients while vitamin C shows non-significant lower values. Regarding the enzymatic antioxidants, both glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly higher in autistic compared to control while catalase recorded more or less similar activities in both groups. CONCLUSION: Saudi autistic children are under H(2)O(2) stress due to GSH depletion, over expression of SOD together with the unchanged catalase enzyme. This could be helpful in the early diagnosis of young autistic patients and suggesting the possibility of antioxidant supplementation for the early intervention with autistic children. PMID- 19306863 TI - Drosophila Fragile X protein controls cellular proliferation by regulating cbl levels in the ovary. AB - FMRP is an RNA binding protein linked to the most common form of inherited mental retardation, Fragile X syndrome (FraX). In addition to severe cognitive deficits, FraX etiology includes postpubescent macroorchidism, which is thought to result from overproliferation. Using a Drosophila FraX model, we show that FMRP controls germline proliferation during oogenesis. dFmr1 null ovaries contain egg chambers with both fewer and supranumerary germ cells. The mutant germaria contain a significantly increased number of cyclin E and PhosphoHistone H3 positive cells, suggesting that loss of FMRP leads to defects in cell cycle progression. BrdU incorporation and flow cytometry data suggest that, in addition to proliferation, germline endoreplication and ploidy are also affected by the loss of FMRP during ovary development. Here we report that FMRP controls the levels of cbl mRNA in the ovary and that reducing cbl gene dosage by half rescues the dFmr1 oogenesis phenotypes. These data support a model whereby FMRP controls germline proliferation by regulating the expression of cbl in the developing ovary. PMID- 19306864 TI - Osa, a subunit of the BAP chromatin-remodelling complex, participates in the regulation of gene expression in response to EGFR signalling in the Drosophila wing. AB - Gene expression is regulated in part by protein complexes containing ATP dependent chromatin-remodelling factors of the SWI/SNF family. In Drosophila there is only one SWI/SNF protein, named Brahma, which forms the catalytic subunit of two complexes composed of different proteins. The protein Osa defines the BAP complex, and the proteins Polybromo and Bap170 are only present in the complex named PBAP. In this work we have analysed the functional requirements of Osa during Drosophila wing development, and found that osa is needed for cell growth and survival in the wing imaginal disc, and for the correct patterning of sensory organs, veins and the wing margin. Other members of the BAP complex, such as Snr1, Bap55, Mor and Brm, also share these functions of Osa. We focused on the requirement of Osa during the formation of the wing veins. Genetic interactions between osa alleles and mutations affecting the activity of the EGFR pathway suggest that one aspect of Osa is intimately related to the response to EGFR activity. Thus, loss of osa and EGFR signalling results in similar wing vein phenotypes, and osa alleles enhance the loss of veins caused by reduced EGFR activity. In addition, Osa is required for the expression of several targets of EGFR signalling, such as Delta, rhomboid and argos. We suggest that one role of Osa and Brm in the wing is to establish a chromatin environment in the regulatory regions of EGFR target genes, making them available for both activators and repressors and facilitating transcription in response to EGFR signalling. PMID- 19306865 TI - Restricted patterns of Hoxd10 and Hoxd11 set segmental differences in motoneuron subtype complement in the lumbosacral spinal cord. AB - During normal vertebrate development, Hoxd10 and Hoxd11 are expressed by differentiating motoneurons in restricted patterns along the rostrocaudal axis of the lumbosacral (LS) spinal cord. To assess the roles of these genes in the attainment of motoneuron subtypes characteristic of LS subdomains, we examined subtype complement after overexpression of Hoxd10 or Hoxd11 in the embryonic chick LS cord and in a Hoxd10 loss-of-function mouse embryo. Data presented here provide evidence that Hoxd10 defines the position of the lateral motor column (LMC) as a whole and, in rostral LS segments, specifically promotes the development of motoneurons of the lateral subdivision of the lateral motor column (LMCl). In contrast, Hoxd11 appears to impart a caudal and medial LMC (LMCm) identity to some motoneurons and molecular profiles suggestive of a suppression of LMC development in others. We also provide evidence that Hoxd11 suppresses the expression of Hoxd10 and the retinoic acid synthetic enzyme, retinaldehyde dehydrogenase 2 (RALDH2). In a normal chick embryo, Hoxd10 and RALDH2 are expressed throughout the LS region at early stages of motoneuron differentiation but their levels decline in Hoxd11-expressing caudal LS segments that ultimately contain few LMCl motoneurons. We hypothesize that one of the roles played by Hoxd11 is to modulate Hoxd10 and local retinoic acid levels and thus, perhaps define the caudal boundaries of the LMC and its subtype complement. PMID- 19306866 TI - The planar cell polarity pathway directs parietal endoderm migration. AB - Parietal endoderm (PE) contributes to the yolk sac and is the first migratory cell type in the mammalian embryo. We can visualize PE migration in vitro using the F9 teratocarcinoma derived embryoid body outgrowth system and, show here that PE migration is directed by the non-canonical Wnt planar cell polarity (PCP) pathway via Rho/ROCK. Based on golgi apparatus localization and microtubule orientation, 68.6% of cells in control outgrowths are oriented in the direction of migration. Perturbation of Wnt signaling via sFRP treatment results in a loss of orientation coupled with an increase in cell migration. Inhibition of the PCP pathway at the level of Daam1 also results in a loss of cell orientation along with an increase in cell migration, as seen with sFRP treatment. Constitutively active Daam can inhibit the loss of orientation that occurs with sFRP treatment. We previously demonstrated that ROCK inhibition leads to an increase in cell migration, and we now show that these cells also lack oriented migration. Canonical Wnt signaling or the Rac arm of the PCP pathway does not appear to play a role in PE oriented migration. These data suggest the PCP pathway via Rho/ROCK modulates migration of PE. PMID- 19306867 TI - Effect of phosphodiesterase antagonists on glucocorticoid mediated growth inhibition in murine skin cell lines. AB - The effects of two cyclic nucleotide phosphodiesterase type 4 (PDE4) inhibitors on proliferation of cell lines representing different stages of mouse skin tumorigenesis were studied. Skin papillomas and carcinomas become resistant to the growth inhibition by glucocorticoids. Their control of cellular functions is mediated by a well-known transcription factor, glucocorticoid receptor. The primary aim of the present study was to determine whether the PDE4 inhibitors, that raise intracellular cAMP levels, can increase the sensitivity of mouse skin papillomas and carcinomas to the glucocorticoids. We sought to establish the effect of cAMP signaling on the glucocorticoid receptor function using well-known model representing non-tumorigenic keratinocyte cell line (3PC), papilloma (MT1/2) and squamous cell carcinoma cell line (Ca3/7). These cells were treated with the glucocorticoid fluocinolone acetonide (FA) alone or in concert with PDE4 inhibitors--rolipram or YM976. Results of our study revealed that both PDE4 inhibitors may increase the sensitivity of transformed cell lines to the growth inhibitory effect of FA. In the transformed cell lines, changes in the viability of cells were accompanied by an increase in mRNA level of two negative regulators of the cell cycle--p21 and p27 proteins. Co-treatment with PDE4 inhibitors and FA caused inhibition of an endogenous glucocorticoid-responsive gene (MT-1) expression. Thus, the PDE4 inhibitors exerted a differential effect on non transformed and transformed keratinocytes and on glucocorticoid receptor signal transduction. These findings warrant further studies to clarify the mechanism by which PDE4 inhibitors modulate glucocorticoid receptor signal transduction in transformed cells. PMID- 19306869 TI - Focus on molecules: ABCA4 (ABCR)--an import-directed photoreceptor retinoid flipase. PMID- 19306868 TI - Sox9 directly promotes Bapx1 gene expression to repress Runx2 in chondrocytes. AB - The transcription factor, Sry-related High Mobility Group (HMG) box containing gene 9 (Sox9), plays a critical role in cartilage development by initiating chondrogenesis and preventing the subsequent maturation process called chondrocyte hypertrophy. This suppression mechanism by Sox9 on late-stage chondrogenesis partially results from the inhibition of Runt-related transcription factor 2 (Runx2), the main activator of hypertrophic chondrocyte differentiation. However, the precise mechanism by which Sox9 regulates late chondrogenesis is poorly understood. In the present study, the transcriptional repressor vertebrate homolog of Drosophila bagpipe (Bapx1) was found to be a direct target of Sox9 for repression of Runx2 expression in chondrocytes. We identified a critical Sox9 responsive region in the Bapx1 promoter via a luciferase reporter assay. Analysis by chromatin immunoprecipitation and electrophoretic mobility shift assays indicated that Sox9 physically bound to this region of the Bapx1 promoter. Consistent with the notion that Bapx1 and Sox9 act as negative regulators of chondrocyte hypertrophy by regulating Runx2 expression, transient knockdown of Sox9 or Bapx1 expression by shRNA in chondrocytes increased Runx2 expression, as well as expression of the late chondrogenesis marker, Col10a1. Furthermore, while over-expression of Sox9 decreased Runx2 and Col10a1 expressions, simultaneous transient knockdown of Bapx1 diminished that Sox9 over-expressing effect. Our findings reveal that the molecular pathway modulated by Bapx1 links two major regulators in chondrogenesis, Sox9 and Runx2, to coordinate skeletal formation. PMID- 19306870 TI - Functional MRI study of the primary somatosensory cortex in comatose survivors of cardiac arrest. AB - It is difficult to assess cerebral function in comatose patients. Because earlier functional neuroimaging studies demonstrate associations between cerebral metabolism and levels of consciousness, fMRI in comatose survivors of cardiac arrest could provide further insight into cerebral function during coma. Using fMRI, cerebral activation to somatosensory stimulation to the palm of the hand was measured in 19 comatose survivors of cardiac arrest and in 10 healthy control subjects and was compared to somatosensory-evoked potential (SSEP) testing of the median nerve. Changes in the blood oxygenation-level dependent signal (BOLD) in the primary somatosensory cortex (S1) contralateral to the stimulated hand were quantified. Clinical outcome was assessed using the Glasgow Outcome Scale (GOS) and the modified Rankin Scale at 3 months post-cardiac arrest. Five out of 19 patients were alive at 3 months. Patients who survived cardiac arrest showed greater BOLD in S1 contralateral to somatosensory stimulation of the hand compared to patients who eventually did not. Greater BOLD was also seen in S1 of patients who retained their SSEP N20 waveforms. There were also positive correlations between BOLD in S1 with both levels of consciousness and measures of outcome at 3 months. In summary, this study demonstrates that BOLD in the S1 contralateral to somatosensory stimulation of the hand varies with clinical measures of the level of consciousness during coma. PMID- 19306871 TI - Neck rigidity in Parkinson's disease patients is related to incomplete suppression of reflexive head stabilization. AB - Muscle rigidity in PD (Parkinson's disease) patients represents an involuntary increase in muscle tone that stands out upon passive rotation of a joint. The pathophysiology of rigidity is still not well understood. We measured head-trunk torque in PD patients and normal controls during transient passive head rotations by means of servomotors under the instruction to the subjects to relax the neck muscles. We observed that rotation onset was followed by an initial rapid rise in resistive torque, similarly in both subject groups. It then leveled off or declined in controls. With PD patients, in contrast, the rise continued roughly proportional to head eccentricity almost until the end of the rotation. These observations led us to the hypothesis that the initial rise in torque represents reflexive head stabilization that normal subjects in the course of the rotational stimulus are able to suppress, whereas PD patients are less effective in doing so. The hypothesis was implemented into a dynamic control model of active and passive head rotation. Model simulations successfully reproduced the torque responses of normal subjects and PD patients in the present and previous studies. PMID- 19306872 TI - Functional relevance of ipsilateral motor activation in congenital hemiparesis as tested by fMRI-navigated TMS. AB - Many, but not all patients with congenital hemiparesis (i.e., hemiparesis due to a pre-, peri- or neonatally acquired brain lesion) control their paretic hands via ipsilateral cortico-spinal projections from the contra-lesional hemisphere (CON-H). Patients who still control their paretic hands via preserved crossed cortico-spinal projections from the damaged hemisphere nevertheless show increased fMRI activation during paretic hand movements in the CON-H. We used fMRI-navigated rTMS induced functional lesions over the primary motor cortex (M1) hand area, the dorsal premotor cortex (dPMC) and the superior parietal lobe (SPL) of the CON-H in four of these patients to investigate whether this increased ipsilateral activation during finger movements of the paretic hand contributes to movement performance. Functional lesions of the dPMC and M1 but not SPL of the CON-H induced decreased temporal preciseness of finger sequences. The present results argue for a possible role of dPMC and M1 of the CON-H on complex motor behavior even in those patients with congenital hemiparesis who control their paretic hands via crossed cortico-spinal projections from the damaged hemisphere. PMID- 19306873 TI - Differentiation-dependent progesterone synthesis and metabolism in NT2-N human neurons. AB - Human embryonic teratocarcinoma-derived Ntera2/cl.D1 (NT2) cells recapitulate many features of embryonic neuronal progenitor cells. Upon retinoic acid (RA) treatment they terminally differentiate into post-mitotic neuron-like cells (NT2 N), akin to human fetal neurons, thus representing an in vitro model of human neuron terminal differentiation. Experimental evidence also indicate NT2-N cultures as a potential source for cell transplantation therapy. The neurosteroids progesterone and its metabolite 3alpha-hydroxy-5alpha-pregnan-20 one (3alpha,5alpha-THP) promote neurogenesis and show anti-neurodegenerative properties. This study's aim was to assess the neurosteroidogenic competence of NT2 cells during RA-induced neuronal differentiation. Radioimmunoassay measurements revealed progesterone only in NT2-N cultures (4 week RA). Accordingly, progesterone synthesis from (3)H-pregnenolone was absent in NT2 cells and increased during RA exposure, being highest in NT2-N. [(3)H] pregnenolone metabolism, yielding [(3)H]-progesterone and [(3)H]-5alpha dihydroprogesterone ([(3)H]-5alpha-DHP), was time-dependent and inhibited by trilostane, a 3beta-hydroxysteroid-dehydrogenase (3beta-HSD) inhibitor. Conversely, (3)H-progesterone metabolism, which yielded [(3)H]-5alpha-DHP > [(3)H]-3beta,5alpha-THP > [(3)H]-3alpha,5alpha-THP, occurred at all time points examined, though showing a nadir in cultures treated with RA for 1 and 2 weeks. The differentiation-dependent increase of progesterone accumulation matched 3beta HSD type I mRNA expression and 3beta-HSD immunoreactivity, that co-localized with Map2a/b- and GAD67 in NT2-N. Hence, in vitro differentiated human neurons, while retaining progesterone metabolic activity, also become competent in progesterone synthesis. These findings suggest an autocrine/paracrine role of neuronal progesterone, either on its own or through its 5alpha-reduced metabolites, in fetal brain development and allow speculation that NT2-N-produced neurosteroids may contribute to the encouraging results of NT2-N transplants in animal models of neurodegenerative diseases. PMID- 19306874 TI - IgM anti-GQ1b monoclonal antibody inhibits voltage-dependent calcium current in cerebellar granule cells. AB - Miller-Fisher syndrome (MFS), which is known to be associated with anti-GQ1b antibodies and to cause ataxia, is a variant of an acute inflammatory neuropathy. However, the pathogenic role of anti-GQ1b antibodies remains unclear. In this study, we investigated the effects of mouse IgM anti-GQ1b monoclonal antibody (IgM anti-GQ1b mAb) on the spontaneous muscle action potential of a rat spinal cord-muscle co-culture system and on the voltage-dependent calcium channel (VDCC) current in cerebellar granule cells and Purkinje cells using the whole-cell patch clamp technique. The frequency of spontaneous muscle action potential of the innervated muscle cells was transiently increased by IgM anti-GQ1b mAb and then was blocked completely, which was the same finding as reported previously. Moreover, the cerebellar granule cell VDCC current was decreased by 30.76+/-7.60% by 5 microg/mL IgM anti-GQ1b mAb, whereas IgM anti-GQ1b mAb did not affect the VDCC current in cerebellar Purkinje cells. In immunocytochemistry, IgM anti-GQ1b mAb stained the whole cell surface of cerebellar granule cells, but not that of Purkinje cells. Therefore, the clinical symptoms of Miller-Fisher syndrome, such as cerebellar-like ataxia, may be explained by the inhibitory effects of anti GQ1b antibodies on VDCC current in cerebellar granule cells. PMID- 19306875 TI - Beta2-chimaerin binds to EphA receptors and regulates cell migration. AB - Ephrins and Eph receptors have key roles in regulation of cell migration during development. We found that the RacGAP beta2-chimaerin (chimerin) bound to EphA2 and EphA4 and inactivated Rac1 in response to ephrinA1 stimulation. EphA4 bound to beta2-chimaerin through its kinase domain and promoted binding of Rac1 to beta2-chimaerin. In addition, knockdown of endogenous beta2-chimaerin blocked ephrinA1-induced suppression of cell migration. These results suggest that beta2 chimaerin is activated by EphA receptors and mediates the EphA receptor-dependent regulation of cell migration. PMID- 19306876 TI - The effect of 2-deoxy-D-glucose on Werner syndrome RecQ helicase gene. AB - Caloric restriction (CR) is known to effectively elongate mammalian life-spans. The compound 2-deoxy-D-glucose (2DG), which is often used as an inhibitor of glucose utilization, is a mimetic agent of CR. In this study, we examined the changes of telomerase and Werner's syndrome RecQ (WRN) helicase after treatment with 2DG, because of the involvement of recQ helicase in the regulation of telomeres. Interestingly, 2DG treatment increased the expression of WRN protein in accordance with induction of its promoter activity and gene expression. Furthermore, the activation of telomerase was observed after 2DG treatment, whereas it resulted in the reduction of cell proliferation. These results suggest that 2DG could up-regulate telomere maintenance factors accompanied with suppression of proliferation. PMID- 19306877 TI - Bioinformatics analysis of mass spectrometry-based proteomics data sets. AB - Proteomics has made tremendous progress, attaining throughput and comprehensiveness so far only seen in genomics technologies. The consequent avalanche of proteome level data poses great analytical challenges for downstream interpretation. We review bioinformatic analysis of qualitative and quantitative proteomic data, focusing on current and emerging paradigms employed for functional analysis, data mining and knowledge discovery from high resolution quantitative mass spectrometric data. Many bioinformatics tools developed for microarrays can be reused in proteomics, however, the uniquely quantitative nature of proteomics data also offers entirely novel analysis possibilities, which directly suggest and illuminate biological mechanisms. PMID- 19306878 TI - Crystal structure of polysaccharide lyase family 20 endo-beta-1,4-glucuronan lyase from the filamentous fungus Trichoderma reesei. AB - The crystal structure of endo-beta-(1-->4)-glucuronan lyase from Trichoderma reesei (TrGL) has been determined at 1.8A resolution as the first three dimensional structure of polysaccharide lyase (PL) family 20. TrGL has a typical beta-jelly roll fold, which is similar to glycoside hydrolase family 16 and PL7 enzymes. A calcium ion is bound to the site far from the cleft and appears to contribute to the stability. There are several completely conserved residues in the cleft. Possible catalytic residues are predicted based on structural comparison with PL7 alginate lyase A1-II'. PMID- 19306879 TI - Focal adhesion disassembly requires clathrin-dependent endocytosis of integrins. AB - Cell migration requires the controlled disassembly of focal adhesions, but the underlying mechanisms remain poorly understood. Here, we show that adhesion turnover is mediated through dynamin- and clathrin-dependent endocytosis of activated beta1 integrins. Consistent with this, clathrin and the clathrin adaptors AP-2 and disabled-2 (DAB2) distribute along with dynamin 2 to adhesion sites prior to adhesion disassembly. Moreover, knockdown of either dynamin 2 or both clathrin adaptors blocks beta1 integrin internalization, leading to impaired focal adhesion disassembly and cell migration. Together, these results provide important insight into the mechanisms underlying adhesion disassembly and identify novel components of the disassembly pathway. PMID- 19306880 TI - Ca2+-modulated ONE-GC odorant signal transduction. AB - In a subset of olfactory epithelium the odorant receptor guanylate cyclase, ONE GC, is a central transduction component of the cyclic GMP signaling pathway. The odorant binds to the extracellular domain and activates its intracellular catalytic domain to generate the odorant second messenger, cyclic GMP. The present study demonstrates that it is a two-step, Ca(2+)-independent and Ca(2+) dependent, sequential process. In step one, the odorant, uroguanylin, binds ONE GC and primes it for stimulation. In step two, Ca(2+)-bound neurocalcin delta binds to the defined intracellular domain and saturates ONE-GC activity. A prototype model is proposed that depicts this signal transduction process. PMID- 19306881 TI - Neural activation in the orbitofrontal cortex in response to male faces increases during the follicular phase. AB - Women's sexual interest changes with hormonal fluctuations across the menstrual cycle. It is unclear how hormones modify women's sexual behavior and desire, but one possibility is that they alter women's positive appraisals of stimuli and thus their sexual interest. Using 3 T fMRI, we measured neural activation in women at two time points in their menstrual cycle (late follicular, luteal) while they evaluated photos of men presented as potential sexual partners. Participants were ten heterosexual women aged 23-28 none of who was using hormonal contraceptives or in a committed relationship. In an event-related design, the women were presented with as series of photos of male faces and asked questions to assess their degree of sexual interest in the men depicted. Results demonstrate an overall effect of menstrual cycle phase on neural activation. During their follicular versus luteal phase, women demonstrated increased activation in the right medial orbitofrontal cortex (OFC), suggesting increased positive appraisal. Activation in the OFC was positively correlated with women's estradiol to progesterone ratios. There were no areas that demonstrated increased activation during the luteal versus follicular phase. The observed increase in activation in the OFC during the follicular phase may reflect a hormonally mediated increase in appetitive motivation and may prime women towards increased sexual interest and behavior around ovulation. PMID- 19306882 TI - Histopathological changes of Ceroplastes japonicus infected by Lecanicillium lecanii. AB - The infection process and pathological changes of Japanese wax scale, Ceroplastes japonicus Green, by the hyphomycete Lecanicillium lecanii (Zimmermann) Gams & Zare were investigated by light, scanning and transmission electron microscopy. The results showed that L. lecanii generally infected the wax scale by penetrating the integument. The anal area, the body margin, around the base of mouthparts and legs, over the stigmatic furrow and the area around the vulva were susceptible places, while the wax test had an inhibitory effect on L. lecanii. Within 24h after inoculation, conidia became attached to the cuticle, and within 48h, hyphae adhered to the integument of the scale and their tips differentiated into specialized infection pegs. Penetration of the cuticle occurred within 72h of inoculation; the fungus caused the insect cuticle to rupture and hyphae entered the insect body through these openings. Within 72h after inoculation, L. lecanii entered the hemocoele of the scale and formed blastospores. After 96h, blastospores were dispersed throughout the hemolymph and completely disrupted the hemocytes, resulting in damage of the cell nucleus and agglutination of chromatin. Concomitant to colonization of the hemolymph, the internal organs and tissues, e.g., tracheae, malpighian tubules and muscle fibers, were also infected. As the infection progressed, the wax test and body changed color from white and red, respectively, to yellowish. After 144h, the internal tissue structure was totally compromised and the insects died. After this time, new conidiophores bearing conidia were produced on the surface of the cadavers. PMID- 19306883 TI - A common interaction for the entry of colicin N and filamentous phage into Escherichia coli. AB - Colicin N is a pore-forming bacteriocin that enters target Escherichia coli cells with the assistance of TolA, a protein in the periplasm of the target cell. The N terminal domain of the colicin that carries the TolA-binding epitope, the translocation domain (T-domain), is intrinsically disordered. From (1)H-(13)C (15)N NMR studies of isotopically labeled T-domain interacting with unlabeled TolAIII (the C-terminal domain of TolA), we have identified the TolA-binding epitope and have shown that the extent of its disorder is reduced on binding TolA, although it does not fold into a globular structure with defined secondary structure elements. Residues upstream and downstream of the 27-residue TolA binding epitope remain disordered in the TolA-bound T-domain as they are in the free T-domain. Filamentous phage also exploits TolAIII to enter target cells, with TolAIII retaining its main secondary structure elements and global fold. In contrast to this, binding of the disordered T-domain of colicin A causes dramatic conformational changes in TolAIII marked by increased flexibility and lack of a rigid tertiary structure consistent with at least partial unfolding of TolAIII, suggesting that bacteriocins and bacteriophages parasitize E. coli using different modes of interaction with TolAIII. We have found that the colicin N T domain-TolAIII interaction is strikingly similar to the previously described g3p TolAIII interaction. The fact that both colicin N and filamentous phage exploit TolAIII in a similar manner, with one being a bacterial intrinsically disordered protein and the other being a viral structurally well-ordered protein, suggests that these represent a good example of convergent evolution at the molecular level. PMID- 19306884 TI - Titin-induced force enhancement and force depression: a 'sticky-spring' mechanism in muscle contractions? AB - The sliding filament and crossbridge theories do not suffice to explain a number of muscle experiments. For example, from the entire muscle to myofibrils, predictions of these theories were shown to underestimate the force output during and after active tissue stretch. The converse applies to active tissue shortening. In addition to the crossbridge cycle, we propose that another molecular mechanism is effective in sarcomere force generation. We suggest that, when due to activation, myosin binding sites are available on actin, the giant protein titin's PEVK region attaches itself to the actin filament at those sites. As a result, the molecular spring length is dramatically reduced. This leads to increased passive force when the sarcomere is stretched and to decreased or even negative passive force when the sarcomere shortens. Moreover, during shortening, the proposed mechanism interferes with active-force production by inhibiting crossbridges. Incorporation of a simple 'sticky-spring' mechanism model into a Hill-type model of sarcomere dynamics offers explanations for several force enhancement and force-depression effects. For example, the increase of the sarcomere force compared to the force predicted solely by the sliding filament and crossbridge theories depends on the stretch amplitude and on the working range. The same applies to the decrease of sarcomere force during and after shortening. Using only literature data for its parameterization, the model predicts forces similar to experimental results. PMID- 19306885 TI - Regulation of kinase activity by diffusion and feedback. AB - In living cells proteins motilities regulate the spatiotemporal dynamics of molecular pathways. We consider here a reaction-diffusion model of mutual kinase receptor activation showing that the strength of positive feedback is controlled by the kinase diffusion coefficient. For high diffusion, the activated kinase molecules quickly leave the vicinity of the cell membrane and cannot efficiently activate the receptors. As a result, in a broad range of parameters, the cell can be activated only if the kinase diffusion coefficient is sufficiently small. Our simple model shows that change in the motility of substrates may dramatically influence the cell responses. PMID- 19306886 TI - Heterogeneity in susceptibility to infection can explain high reinfection rates. AB - Heterogeneity in susceptibility and infectivity is inherent to infectious disease transmission in nature. Here we are concerned with the formulation of mathematical models that capture the essence of heterogeneity while keeping a simple structure suitable of analytical treatment. We explore the consequences of host heterogeneity in the susceptibility to infection for epidemiological models for which immunity conferred by infection is partially protective, known as susceptible-infected-recovered-infected (SIRI) models. We analyze the impact of heterogeneity on disease prevalence and contrast the susceptibility profiles of the subpopulations at risk for primary infection and reinfection. We present a systematic study in the case of two frailty groups. We predict that the average rate of reinfection may be higher than the average rate of primary infection, which may seem paradoxical given that primary infection induces life-long partial protection. Infection generates a selection mechanism whereby fit individuals remain in S and frail individuals are transferred to R. If this effect is strong enough we have a scenario where, on average, the rate of reinfection is higher than the rate of primary infection even though each individual has a risk reduction following primary infection. This mechanism may explain high rates of tuberculosis reinfection recently reported. Finally, the enhanced benefits of vaccination strategies that target the high-risk groups are quantified. PMID- 19306887 TI - Catastrophic regime shifts in coral communities exposed to physical disturbances: simulation results from object-oriented 3-dimensional coral reef model. AB - A 3-dimensional individual-based model, the ReefModel, was developed to simulate the dynamical structure of coral reef community using object-oriented techniques. Interactions among functional groups of reef organisms were simulated in the model. The behaviours of these organisms were described with simple mechanistic rules that were derived from their general behaviours (e.g. growing habits, competitive mechanisms, response to physical disturbance) observed in natural coral reef communities. The model was implemented to explore the effects of physical disturbance on the dynamical structure of a 3-coral community that was characterized with three functional coral groups: tabular coral, foliaceous coral and massive coral. Simulation results suggest that (i) the integration of physical disturbance and differential responses (disturbance sensitivity and growing habit) of corals plays an important role in structuring coral communities; (ii) diversity of coral communities can be maximal under intermediate level of acute physical disturbance; (iii) multimodality exists in the final states and dynamic regimes of individual coral group as well as coral community structure, which results from the influence of small random spatial events occurring during the interactions among the corals in the community, under acute and repeated physical disturbances. These results suggest that alternative stable states and catastrophic regime shifts may exist in a coral community under unstable physical environment. PMID- 19306888 TI - Platelets and PEGylated lecithin liposomes: when stealth is allegedly picked up on the radar (and eaten). PMID- 19306889 TI - Satratoxin G interaction with 40S and 60S ribosomal subunits precedes apoptosis in the macrophage. AB - Satratoxin G (SG) and other macrocyclic trichothecene mycotoxins are potent inhibitors of eukaryotic translation that are potentially immunosuppressive. The purpose of this research was to test the hypothesis that SG-induced apoptosis in the macrophage correlates with binding of this toxin to the ribosome. Exposure of RAW 264.7 murine macrophages to SG at concentrations of 10 to 80 ng/ml induced DNA fragmentation within 4 h that was indicative of apoptosis. To relate these findings to ribosome binding of SG, RAW cells were exposed to different toxin concentrations for various time intervals, ribosomal fractions isolated by sucrose density gradient ultracentrifugation and resultant fractions analyzed for SG by competitive ELISA. SG was found to specifically interact with 40S and 60S ribosomal subunits as early as 5 min and that, at high concentrations or extended incubation times, the toxin induced polysome disaggregation. While co-incubation with the simple Type B trichothecene DON had no effect on SG uptake into cell cytoplasm, it inhibited SG binding to the ribosome, suggesting that the two toxins bound to identical sites and that SG binding was reversible. Although both SG and DON induced mobilization of p38 and JNK 1/2 to the ribosome, phosphorylation of ribosomal bound MAPKs occurred only after DON treatment. SG association with the 40S and 60S subunits was also observed in the PC-12 neuronal cell model which is similarly susceptible to apoptosis. To summarize, SG rapidly binds small and large ribosomal subunits in a concentration- and time-dependent manner that was consistent with induction of apoptosis. PMID- 19306890 TI - A hydra with many heads: protein and polypeptide toxins from hydra and their biological roles. AB - Hydra have been classical model organisms for over 250 years, yet little is known about the toxins they produce, and how they utilize these toxins to catch prey, protect themselves from predators and fulfill other biological roles necessary for survival. Unlike typical venomous organisms the hydra allomonal system is complex and "holistic", produced by various stinging cells (in the hunting tentacles and body ectoderm) as well as by non-nematocystic tissue. Toxic proteins also fulfill novel, non-allomonal roles in hydra. This review described the toxins produced by hydra within the context of their biology and natural history. Hydra nematocyst venom contains a high-molecular weight (>100 kDa) hemolytic and paralytic protein and a protein of approximately 30 kDa which induces a long-lasting flaccid paralysis. No low-molecular weight toxicity is observed, suggesting the lack of "classical" 4-7 kDa neurotoxins. The occurrence of a potent phospholipase activity in the venom is supported by the detection of several venom-like phospholipase A2 genes expressed by hydra. Hydra also produce toxins which are not part of the nematocyst venom. In the green hydra, Hydralysins, a novel family of Pore-Forming Proteins, are secreted into the gastrovascular cavity during feeding, probably helping in disintegration of the prey. Other putative non-nematocystic "toxins" may be involved in immunity, development or regulation of behavior. As the first venomous organism for which modern molecular tools are available, hydra provide a useful model to answer many outstanding questions on the way venomous organisms utilize their toxins to survive. PMID- 19306891 TI - Chemical synthesis and folding of APETx2, a potent and selective inhibitor of acid sensing ion channel 3. AB - Acid sensing ion channels (ASICs) are pH-sensitive channels that are distributed in the central and peripheral nervous system and which are believed to play a key role in pain perception. APETx2, a 42-residue peptide toxin isolated from the sea anemone Anthopleura elegantissima, is the only known selective inhibitor of ASIC3 channels. Here we describe the total chemical synthesis of APETx2 by solid-phase peptide synthesis and native chemical ligation. The folded synthetic toxin had an IC(50) of 57 nM for inhibition of rat ASIC3 channels expressed in Xenopus oocytes, in agreement with the IC(50) reported for the native toxin (63 nM). The native chemical ligation approach should provide an efficient route for synthesis of other pharmacologically useful disulfide-rich toxins from venomous animals. PMID- 19306892 TI - Effects of luminance contrast and its modifications on fixation behavior during free viewing of images from different categories. AB - During viewing of natural scenes, do low-level features guide attention, and if so, does this depend on higher-level features? To answer these questions, we studied the image category dependence of low-level feature modification effects. Subjects fixated contrast-modified regions often in natural scene images, while smaller but significant effects were observed for urban scenes and faces. Surprisingly, modifications in fractal images did not influence fixations. Further analysis revealed an inverse relationship between modification effects and higher-level, phase-dependent image features. We suggest that high- and mid level features--such as edges, symmetries, and recursive patterns--guide attention if present. However, if the scene lacks such diagnostic properties, low level features prevail. We posit a hierarchical framework, which combines aspects of bottom-up and top-down theories and is compatible with our data. PMID- 19306893 TI - Smoking-related knowledge, attitudes and behaviors in the lesbian, gay and bisexual community: a population-based study from the U.S. Pacific Northwest. AB - OBJECTIVE: Several studies have shown that lesbian, gay and bisexual (LGB) persons have higher smoking prevalence than heterosexuals. However, few population-based studies have explored whether smoking-related knowledge, attitudes and behaviors also differ between the communities. METHODS: We used Behavioral Risk Factor Surveillance System data for 2003 to 2005 from two states (Washington and Oregon) to compare smoking-related indicators between the self identified LGB population and their heterosexual counterparts. RESULTS: Lesbians, gays and bisexuals were more likely to be current or ever smokers than their heterosexual counterparts. All except bisexual men and had lower quit ratios than heterosexuals. Among successful quitters, bisexual men were less likely to be long-term quitters than heterosexuals. For all groups, attitudes and behaviors regarding secondhand smoke (SHS) were similar to those of heterosexuals, except for bisexual women, who were more likely to be exposed to SHS. CONCLUSIONS: Despite a disparity in smoking prevalence, the LGB population in these two states appeared to have similar levels of knowledge and attitudes toward tobacco control as their heterosexual counterparts. Nevertheless, tobacco control programs should continue to focus on this population to prevent smoking initiation, promote cessation, and reduce secondhand smoke exposure. PMID- 19306894 TI - Target site occupancy: emerging generalizations from clinical and preclinical studies. AB - What percentage of receptors, ion channels or transporters must be occupied by drugs to trigger therapeutic effects in patients, or by drugs and other ligands to induce physiological effects in humans or animals? Human studies utilizing Positron Emission Tomography and Single Photon Emission Computed Tomography, along with data from an array of preclinical methodologies, have begun to provide consistent answers to this question. The required target occupancy is dependent upon the molecular class of both target and ligand, and appears to be similar for both patient therapy and human or animal physiology. In the case of antagonists, approximately 60-90% target occupancy is required for G protein-coupled receptors, neurotransmitter transporters, and ligand-gated ion channels. Effective doses of agonists occupy a wider range of their target sites, dependent upon the intrinsic activity of the agonist, the receptor or ion channel reserve of the target site, and the response that is measured, with low efficacy agonists generally requiring high degrees of occupancy while high efficacy agonists generally require low degrees of occupancy. Target desensitization, competition by endogenous ligands, and regional target differences all influence target occupancy requirements. Measurements of target occupancy can help assure proper dosing and targeting of compounds in preclinical and clinical drug development as well as in basic research. Target occupancy generalizations can be especially important in establishing initial dosing recommendations for the many new drug targets provided by genomic and proteomic initiatives, where little data is available on their functional responses. PMID- 19306896 TI - Multilevel targeting of hematopoietic stem cell self-renewal, differentiation and apoptosis for leukemia therapy. AB - Human leukemias are considered clonal hematological malignancies initiated by chromosomal aberrations or epigenetic alterations occurring at the level of either pluripotent hematopoietic stem cells (HSCs) or early multipotent progenitors (MPPs). Leukemic cells are transformed, immortalized, actively proliferating cells that are still able to differentiate into cells resembling mature blood cells. Future therapies of leukemias require identification of molecular targets involved in hematopoiesis under normal and leukemic conditions and detailed understanding of the interactions between normal hematopoietic and leukemic cells within the bone marrow micro-environment. This review presents the basic aspects of hematopoiesis and highlights multilevel exploitable targets for leukemia therapy. These include HSC niche components, signaling pathways (SCF/c kit-R, EPO-R-JAK2/STAT, Wnt, Notch, HOX), inducer-receptor interactions, superfine chromatin structure modifications, fused transcription factors, microRNAs and signaling of cell death through the Bcl-2 apoptotic switch (BH3 only proteins). The classes of therapeutics developed or being under development to eradicate human leukemias include novel antimetabolites, DNA hypomethylating agents, histone deacetylation inhibitors (HDACIs), retinoids and other inducers of differentiation, targeted monoclonal antibodies raised against cell surface proteins, pro-apoptotic receptor agonists (PARAs), BH3 peptidomimetics, cell cycle inhibitors, siRNAs and perhaps microRNAs. Some of these agents induce terminal differentiation while others promote cell cycle arrest and apoptosis in leukemia cells. At last but not least, this article describes the mechanisms of removal of damaged/harmful cells from organs since impairment in clearance of such cells can lead to autoimmune disorders by self-antigens. PMID- 19306897 TI - Neurotrophins and neuronal migration in the developing rodent brain. AB - Neurotrophins are known to be key regulators of neuronal survival, differentiation, function and plasticity in the developing and adult rodent brain. A novel role for neurotrophins has been emerging from recent research, that of motogenic and chemoattractant factors for several populations of migrating neuronal precursors in the developing mouse brain. The aim of the present article is to summarize and discuss the studies that have contributed to the existing body of evidence. PMID- 19306895 TI - Cyclic GMP signaling in cardiovascular pathophysiology and therapeutics. AB - Cyclic guanosine 3',5'-monophosphate (cGMP) mediates a wide spectrum of physiologic processes in multiple cell types within the cardiovascular system. Dysfunctional signaling at any step of the cascade - cGMP synthesis, effector activation, or catabolism - have been implicated in numerous cardiovascular diseases, ranging from hypertension to atherosclerosis to cardiac hypertrophy and heart failure. In this review, we outline each step of the cGMP signaling cascade and discuss its regulation and physiologic effects within the cardiovascular system. In addition, we illustrate how cGMP signaling becomes dysregulated in specific cardiovascular disease states. The ubiquitous role cGMP plays in cardiac physiology and pathophysiology presents great opportunities for pharmacologic modulation of the cGMP signal in the treatment of cardiovascular diseases. We detail the various therapeutic interventional strategies that have been developed or are in development, summarizing relevant preclinical and clinical studies. PMID- 19306899 TI - Penetration and efficacy of VEGF siRNA using polyelectrolyte complex micelles in a human solid tumor model in-vitro. AB - A polyelectrolyte complex(PEC) micelle-based siRNA delivery system has been developed for vascular endothelial growth factor (VEGF), and its antitumor efficacy has been demonstrated using in-vivo animal models. Penetration and distribution through the avascular regions of human solid tumors after extravasation are important issues for antitumor efficacy, especially for macromolecules such as VEGF siRNA PEC micelles. Using an in-vitro solid tumor model, multicellular layers(MCL) culture of human colorectal cancer cells, we evaluated the penetration kinetics and efficacy of VEGF siRNA PEC micelles(PEC siRNA) in comparison to unmodified siRNA(N-siRNA). The PEC-siRNA showed full penetration (15-17 layers of cells) with a unique punctuated distribution pattern at 48 h following initial accumulation in the top layers and a significant suppression of mRNA and protein expression in a dose-dependent manner after 72 h exposure. Although the initial penetration of N-siRNA was faster than that of PEC siRNA, N-siRNA showed complete loss of activity due to its instability within 24 h. Our data support the idea that PEC micelle formulation may provide stable penetration tool through the multilayers of cancer cells and ensure the gene silencing effect of VEGF. This study also demonstrated that MCL could serve as a useful in-vitro model to evaluate the dose- and time-dependent profiles of penetration and efficacy of macromolecular delivery systems in human solid tumor avascular regions. PMID- 19306898 TI - Syk and pTyr'd: Signaling through the B cell antigen receptor. AB - The B cell receptor (BCR) transduces antigen binding into alterations in the activity of intracellular signaling pathways through its ability to recruit and activate the cytoplasmic protein-tyrosine kinase Syk. The recruitment of Syk to the receptor, its activation and its subsequent interactions with downstream effectors are all regulated by its phosphorylation on tyrosine. This review discusses our current understanding of how this phosphorylation regulates the activity of Syk and its participation in signaling through the BCR. PMID- 19306900 TI - Targeted lipid-coated nanoparticles: delivery of tumor necrosis factor functionalized particles to tumor cells. AB - Polymeric nanoparticles displaying tumor necrosis factor on their surface (TNF nanocytes) are useful carrier systems capable of mimicking the bioactivity of membrane-bound TNF. Thus, TNF nanocytes are potent activators of TNF receptor 1 and 2 leading to a striking enhancement of apoptosis. However, in vivo applications are hampered by potential systemic toxicity. Here, using TNF nanocytes as a model system, we developed a procedure to generate targeted lipid coated particles (TLP) in which TNF activity is shielded. The TLPs generated here are composed of an inner single-chain TNF (scTNF)-functionalized, polymeric nanoparticle core surrounded by a lipid coat endowed with polyethylene glycol (PEG) for sterical stabilization and a single-chain Fv (scFv) fragment for targeting. Using a scFv directed against the tumor stroma marker fibroblast activation protein (FAP) we show that TLP and scTNF-TLP specifically bind to FAP expressing, but not to FAP-negative cells. Lipid coating strongly reduced nonspecific binding of particles and scTNF-mediated cytotoxicity towards FAP negative cells. In contrast, an increased cytotoxicity of TLP was observed for FAP-positive cells. Thus, through liposome encapsulation, nanoparticles carrying bioactive molecules, which are subject to nonselective uptake and activity towards various cells and tissues, can be converted into target cell-specific composite particles exhibiting a selective activity towards antigen-positive target cells. Besides safe and targeted delivery of death ligands such as TNF, TLP should be suitable for various diagnostic and therapeutic applications, which benefit from a targeted delivery of reagents embedded into the particle core or displayed on the core particle surface. PMID- 19306901 TI - Enzymatically degradable temperature-sensitive polypeptide as a new in-situ gelling biomaterial. AB - We are reporting a poly (ethylene glycol)-block-poly(alanine-co-phenyl alanine) (PEG-PAF) aqueous solution that undergoes sol-to-gel transition as the temperature increases. The sol-to-gel transition was observed at as low a concentration as 3.0-7.0 wt.%. Micellar aggregation accompanying small conformational changes of the peptide from random coils to beta-sheets is suggested as the sol-to-gel transition mechanism of the PEG-PAF aqueous solution. The PEG-PAF is stable in phosphate buffered saline, however, it degraded in the subcutaneous layer of rats. In vitro study showed that proteolytic enzymes such as cathepsin B, cathepsin C, and elastase that are present in the subcutaneous layer of the mammalian tissue might be responsible for the degradation of the polymer in rats. As a feasibility study of this material, a single shot of an aqueous insulin formulation (13.8 mg insulin/kg) showed a hypoglycemic effect over 18 days in rats. The current functional polypeptide may be very promising as an in-situ gelling system for tissue engineering, cell/stem cell therapy, and drug delivery. PMID- 19306902 TI - Anti-oxidants for therapeutic use: why are only a few drugs in clinical use? AB - It is well known that ROS are involved in a wide spectrum of diseases. In spite of this fact, only a few drugs are currently in use in clinical settings: Ebselen for the treatment of cerebral infarctions, N-acetylcysteine for over-doses of acetaminophen. Because ROS production is very common and the human body has an abundant capacity for producing anti-oxidants, it is difficult to obtain statistical differences in trials designed to evaluate an anti-oxidative drug. Additional resources will be needed, if we are to use anti-oxidants in human clinical settings; Drug delivery to a specific region, appropriate setting of evaluation goals in clinical trials, new approaches to understanding the details of anti-oxidative systems. In spite of these drawbacks, the additional use of anti-oxidants in clinical settings is clearly warranted, and could contribute positively to human health. It is generally thought that reactive oxygen species (ROS) are involved in a wide variety of diseases, including ischemia-reperfusion, cancer and various types of inflammation. Because of this, the therapeutic use of antioxidants is now widespread. Some of these have been shown to be successful, but many appear to have no or little benefit, in terms of being beneficial to general health or in disease prevention. In this commentary, the author takes a look back at important anti-oxidative drugs and wonders why only a few such drugs are currently in clinical use. PMID- 19306903 TI - The role of developmental toxicity studies in acute exposure assessments: analysis of single-day vs. multiple-day exposure regimens. AB - In accordance with most toxicity guidelines, developmental studies typically utilize repeated exposures, usually throughout gestation or during organogenesis in particular. However, it is known that developmental toxicity may occur in response to single exposures, especially during specific windows of susceptibility. An overview of the available literature gave sufficient evidence that for many agents, the same endpoints observed in repeated dose, multiple-day studies were also observed in single-day exposures, thus indicating the relevance of developmental toxicity to health assessments of acute exposures. Further, results of benchmark dose modeling of developmental endpoints indicated that for embryo lethality, single-day exposures required a two- to fourfold higher dose than the multiple-day exposures to produce the same level of response. For fused sternebrae, exposures on specific days produced equivalent levels of response at doses that were more similar to those utilized in the repeated exposures. Appreciable differences in biological half-life (and corresponding dose metrics) as well as specific windows of susceptibility may partially explain the observed multiple- vs. single-day exposure dose-response relationships. Our results highlight the need of a more thorough evaluation of outcomes from repeated dose developmental toxicity studies in regards to their importance to chronic and acute risk assessments. PMID- 19306904 TI - Disruptions in spatial working memory, but not short-term memory, induced by repeated ketamine exposure. AB - Treatment with non-competitive N-methyl-D-aspartate (NMDA) antagonists such as phencyclidine or ketamine have been shown to induce schizophrenia-like psychotic and cognitive symptoms in humans and animals. However, there have been a number of contradictory findings regarding the effects of repeated treatment with these drugs on working memory in experimental animals. We hypothesized that processes dependent on dopamine transmission in the medial prefrontal cortex (PFC) may be more sensitive to disruption following these treatment. We assessed the effects of repeated treatment with ketamine on working memory performance using a delayed spatial win-shift procedure conducted on a radial-arm maze, dependent on a neural circuit linking hippocampal and dopamine inputs to the medial PFC. Rats were trained on the task prior to drug exposure, after which they were subjected to one of two dosing regimes of ketamine (30 mg/kg twice a day for either 5 or 10 days). After a 10 day withdrawal period, they were re-tested on the task for 15 days. Ketamine treatment for 10 days, but not 5 days, increased the number of errors and days to re-achieve the criterion on the delayed task. However, in a separate group of rats, subchronic ketamine treatment (10 days) did not affect performance of the non-delayed random foraging task, dependent on the hippocampus, but not the PFC. These results indicate that working memory performance assessed with these procedures is sensitive to disruption following repeated exposure to ketamine. Impairments in working memory induced by these treatments are not attributable to dysfunction of motivational, motor, short-term or spatial memory processes. The use of these procedures may prove useful in modeling impairments in this executive function observed in schizophrenia. PMID- 19306905 TI - Inverse relationship between thalamic and orbitofrontal volumes in obsessive compulsive disorder. AB - Volumetric magnetic resonance imaging (MRI) studies in obsessive-compulsive disorder (OCD) have reported a smaller volume of the orbitofrontal cortex (OFC) and a larger volume of the thalamus compared with healthy controls. Both of these brain regions are strongly connected; therefore, it may be hypothesized that cortical and thalamic alterations are related. Here, we investigated the relationship between thalamic and orbitofrontal volumes in OCD patients relative to healthy controls. MRI volumetric measurements of the thalamus and the OFC were obtained in 16 OCD patients without comorbidity and 16 comparison subjects matched for age, sex and educational level. Partial correlation analyses that controlled for intracranial volume (ICV) were performed to explore relationships between thalamic and OFC volumes in each group. In order to assess the specificity of this relationship, we conducted similar analyses of the anterior cingulate cortex (ACC) as a non-OFC cortical volume. Finally, by using data from previously published volumetric MRI studies, we conducted a meta-regression to explore the relationships between volume changes in these regions of interest. Results showed that thalamic volumes were significantly negatively correlated with OFC volumes in OCD patients (r=-0.83, p<0.001), but not in healthy subjects (r=-0.15, p=0.59). A significant relationship between thalamic and ACC volumes was found neither in the OCD patients (r=0.03, p=0.91) nor in the comparison subjects (r=-0.23, p=0.40). Furthermore, meta-regression analyses showed that previously reported volume changes in the thalamus were significantly correlated with OFC volume changes (r=-0.71, p<0.05), but not with ACC volume changes (r=0.07, p=0.86). Although our results do not allow for any causal relationship to be established, they suggest that structural alterations of both the thalamus and the OFC are inversely and specifically related in OCD. PMID- 19306906 TI - Protective effect of quince (Cydonia oblonga Miller) fruit against oxidative hemolysis of human erythrocytes. AB - The aim of this study was to determine the phenolic content and evaluate the antioxidant activity of quince (Cydonia oblonga) fruit. For this purpose, fruits were separated into pulps, peels and seeds and methanolic extracts were prepared. The phenolic profiles were determined by HPLC/UV and antioxidant properties were studied for their ability to quench the stable free radical 2,2'-diphenyl-1 picrylhydrazyl (DPPH) and to inhibit the 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative hemolysis of human erythrocytes. The main phenolic compounds were 5-O-caffeoylquinic acid for pulp and peel (57% and 29%, respectively) and stellarin-2 for seed (18%). Total phenolics content was 2.5, 6.3 and 0.4g/kg of methanolic extract for pulp, peel and seed, respectively. Pulp and peel extracts showed similar DPPH free radical scavenging activities (EC(50) of 0.6 and 0.8 mg/ml, respectively), while seed extract presented much lower antioxidant potential (EC(50) of 12.2mg/ml). Under the oxidative action of AAPH, pulp and peel extracts showed significant protection of the erythrocyte membrane from hemolysis, in a time- and concentration-dependent manner. Seed extracts by themselves induced extensive hemolysis. These results indicate higher antioxidant activity for certain parts of quince fruit, namely pulp and peel, that may therefore represent accessible sources of natural antioxidants with potential application in nutritional/pharmaceutical fields, as preventive or therapeutic agents in diseases in which free radicals are implicated. PMID- 19306907 TI - Protective role of vitamin E on nickel and/or chromium induced oxidative stress in the mouse ovary. AB - In the present study, we report the invivo effects of nickel chloride (NiCl2; 8 and 16 mg/kg body weight) and/or potassium dichromate (K2Cr2O7; 5 and 10mg/kg body weight) in the ovary of adult mice. The protective role of vitamin E (2mg/kg body weight) along with their combination was also studied. Nickel and/or chromium to mice enhanced the levels of lipid peroxides in the ovary, which was accompanied by a significant decline in the levels of protein, glutathione, total ascorbic acid and activities of superoxide dismutase and catalase. Supplementation of vitamin E along with NiCl2 + K2Cr2O7 significantly lowered the levels of lipid peroxidation and enhanced the antioxidant status. Findings of the present study suggest that vitamin E exerts its protective effect against nickel and/or chromium induced toxicity by preventing lipid peroxidation and protecting antioxidant system in the mouse ovary. PMID- 19306908 TI - Human exposure to insecticide products containing pyrethrins and piperonyl butoxide (2001-2003). AB - Pyrethrum, used as an insecticide for centuries, is derived from dried and ground flowers of Chrysanthemum cinerariaefolium. Its current major use is in insecticide products to the control insects in the home and food handling establishments. We investigated human incidents reported through the American Association of Poison Control Centers (AAPCC) Toxic Exposure Surveillance System (TESS) associated with regulated insecticides containing pyrethrins and piperonyl butoxide (PY/PBO) from 2001 to 2003. Special attention was paid to dermal and respiratory effects. Although there are limitations associated with TESS data, we observed that In view of their widespread use, the data indicates that PY/PBO products can be used with a relatively low risk of adverse effects. Moreover, the data suggest that they are not likely to cause reactions in people with asthma or allergies. PMID- 19306909 TI - Aroclor 1254 impairs spermatogenesis and induces oxidative stress in rat testicular mitochondria. AB - Aroclor 1254 (A1254) has been shown to have potential testicular toxicity. The mechanism of action of A1254 on male reproduction is not clear. The present study was designed to investigate the potential toxicity of A1254 on rat spermatogenesis. Oxidative stress was also assessed in testicular mitochondria as an underlying mechanism. Adult male Wistar rats were injected with A1254 (0, 0.75, 1.5 or 3mg/kg/day i.p.) or with vehicle (corn oil) for 20 consecutive days. A1254 at doses of 1.5 and 3mg/kg/day resulted in a significant decrease in body weight, testes weight, epididymal and relative epididymal weight. Similarly, the relative testis weight was significantly decreased at 3mg/kg/day. Sperm count, motility and daily sperm production were significantly decreased at 1.5 and 3mg/kg/day. The same two doses significantly inhibited the activities of testicular mitochondrial CAT, GPx and GR while the activity of SOD was significantly decreased by 0.75, 1.5 and 3mg/kg/day. The levels of H(2)O(2) generation and LPO were significantly increased in mitochondria in a dose-related pattern. GSH and Vit C were significantly decreased at 0.75, 1.5 and 3mg/kg/day. In conclusion, A1254 impairs spermatogenesis as evidenced, at least partly, by induction of oxidative stress in testicular mitochondria. PMID- 19306910 TI - In vitro cancer chemopreventive properties of polysaccharide extract from the brown alga, Sargassum latifolium. AB - Polysaccharides of edible algae attracted extensive interest due to their numerous biological activities. Sargassum latifolium (Turner) C. Agardh, belongs to Sargassaceae, is a brown algae in red sea shores in Egypt. This work is a novel attempt to explore the cancer chemopreventive activity of different fractions of water-soluble polysaccharide extract derived from S. latifolium. Estimation of cancer chemopreventive activity, specifically anti-initiation, including the modulation of carcinogen metabolism and the antioxidant capacity, revealed that E1 and E4 were potent anti-initiators, where they lead not only to an inhibition in the carcinogen activator cytochrome P450 1A (IC50 2.54 and 10.30 microg/ml, respectively), but also to an induction in the carcinogen detoxification enzymes glutathione-S-transferases (144% and 225% of the control, respectively). E1 and E4 inhibited 59% and 63% of the induced-DNA damage, as measured by comet assay. Similarly both E1 and E4 possessed potential anti promoting properties as indicated by their anti-inflammatory activity. E1 and E4 enhanced the macrophage proliferation; however they dramatically inhibited the stimulated NO (30.7% and 59.3%), TNF-alpha (38.2% and 54.9) and COX-2 (20% and 18%), respectively. E3 showed a selective cytotoxicity against lymphoblastic leukemia (1301 cells), while other fraction extracts had no cytotoxic effect against all tested cell lines. E3 led to a major disturbance in cell cycle including arrest in both S-phases in 1301 cells. This disturbance was associated with an induced-cell death due to apoptosis, but not necrosis. In conclusion, E1 and E4 are promising cancer chemopreventive fractions, since they had tumor anti- initiating activity via their protective modulation of carcinogen metabolism, and tumor anti-promoting activity via their anti-inflammatory activity, while E3 can be considered as a promising anti-cancer agent against leukemia. PMID- 19306911 TI - Circulating hormone adrenomedullin and its binding protein protect neural cells from hypoxia-induced apoptosis. AB - BACKGROUND: Brain ischemia is the underlying cause of neuron death during stroke and brain trauma. Neural cells exposed to ischemia can undergo apoptosis. Adrenomedullin (AM) in combination with its enhancing binding protein, AMBP-1, has been shown to reduce tissue damage in inflammation. METHODS: To evaluate a beneficial effect of AM/AMBP-1 administration in brain ischemia, we employed an in vitro model of neuronal hypoxia using differentiated human neuroblastoma SH SY5Y cells. RESULTS: After exposure to 1% O(2) for 20 h, neural cells were injured with decreased ATP levels and increased LDH release. Pre-administration of AM/AMBP-1 significantly reduced hypoxia-induced cell injury. Moreover, AM/AMBP 1 treatment reduced the number of TUNEL-positive cells and activation of caspase 3, compared to cells exposed to hypoxia alone. AM/AMBP-1 prevented a reduction of cAMP levels and protein kinase A (PKA) activity in neural cells after hypoxia exposure. Correspondingly, an elevation of cAMP levels by forskolin protected neural cells from hypoxia-induced injury. Inhibition of PKA by KT5720 abolished the protective effect of AM/AMBP-1 on hypoxia-induced apoptosis. CONCLUSIONS: AM/AMBP-1 elevates cAMP levels, followed by activating PKA, to protect neural cells from the injury caused by hypoxia. GENERAL SIGNIFICANCE: AM/AMBP-1 may be used as therapeutic agents to prevent neuron damage from brain ischemia. PMID- 19306912 TI - Epithelial-mesenchymal transition in cancer metastasis: mechanisms, markers and strategies to overcome drug resistance in the clinic. AB - Epithelial-mesenchymal transition (EMT) is a key step during embryogenesis. Accumulating evidence suggests a critical role in cancer progression, through which tissue epithelial cancers invade and metastasise. Cell characteristics are highly affected during EMT, resulting in altered cell-cell and cell-matrix interactions, cell motility and invasiveness. Nevertheless, the demonstration of this process in human cancer has been proven difficult and controversial. Besides the fact that the acquisition of mesenchymal characteristics is not a prerequisite for cell migration/invasion, it is a transient event that concerns only few cells in a tumour mass. The induction of EMT depends on the tumour type and its genetic alterations as well as on its interaction with the extracellular matrix. In parallel, trials for EMT identification in clinical samples lack of a widely accepted methodology, nomenclature and reliable markers. This review summarizes the main EMT characteristics and proposes methodologies for better analysis in vitro. It also highlights recent studies identifying cells with EMT characteristics in human cancer and proposes certain markers to identify them in tumour samples. Finally, it cites the recent literature concerning the mechanisms of drug resistance related to EMT in the context of anti-tumour therapies and proposes related new targets for therapy. PMID- 19306913 TI - Compartmentation of the cerebellar nuclei of the mouse. AB - The cerebellar nuclei integrate inhibitory input from Purkinje cells with excitatory input from mossy and climbing fiber collaterals and are the sole cerebellar output. Numerous studies have shown that the cerebellar cortex is highly compartmentalized into hundreds of genetically determined, reproducible topographic units--transverse zones and parasagittal stripes--that can be identified through the expression patterns of numerous molecules. The Purkinje cell stripes project to the cerebellar nuclei. However, there is no known commensurate topographic complexity in the cerebellar nuclei. Rather, conventional anatomical descriptions identify four major subdivisions--the medial, anterior and posterior interposed, and lateral nuclei--together with a few intranuclear subdivisions. To begin to address the apparent complexity gap, we have used a panel of antigens and transgenes to reveal a reproducible molecular heterogeneity in the mouse cerebellar nuclei. Based on the differential expression patterns, singly and in combination, a new cerebellar nuclear topographic map has been constructed. This reveals the subdivision of the cerebellar nuclei into at least 12 reproducible expression domains. We hypothesize that such heterogeneity is the counterpart of the zones and stripes of the cerebellar cortex. PMID- 19306914 TI - A new dicoumarin and anticoagulant activity from Viola yedoensis Makino. AB - A new dicoumarin, named as dimeresculetin (1), together with another dicoumarin, euphorbetin (2) and esculetin (3) were isolated from the ethyl acetate extract of the dried whole plants of Viola yedoensis Makino. The structure of 1 was elucidated as 7-hydroxy-6-[(6,7-dihydroxy-2-oxo-2H-1-benzopyran-5-yl)oxy]-2H-1 benzopyran-2-one on the basis of extensive NMR, as well as the other spectral analysis. Compounds 1-3 exhibited anticoagulant activities with respect to activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT). PMID- 19306915 TI - Complete mitochondrial genome of the sea cucumber Apostichopus japonicus (Echinodermata: Holothuroidea): the first representative from the subclass Aspidochirotacea with the echinoderm ground pattern. AB - Complete mitochondrial genome plays an important role in the accurate revelation of phylogenetic relationships among metazoans. Here we present the complete mitochondrial genome sequence from a sea cucumber Apostichopus japonicus (Echinodermata: Holothuroidea), which is the first representative from the subclass Aspidochirotacea. The mitochondrial genome of A. japonicus is 16,096 bp in length. The heavy strand consists of 31.8% A, 20.2% C, 17.9% G, and 30.1% T bases (AT skew=0.027; GC skew=0.062). It contains thirteen protein-coding genes (PCGs), twenty-two transfer RNA genes, and two ribosomal RNA genes. There are a total of 3793 codons in all thirteen mitochondrial PCGs, excluding incomplete termination codons. The most frequently used amino acid is Leu (15.77%), followed by Ser (9.73%), Met (8.62%), Phe (7.94%), and Ala (7.28%). Intergenetic regions in the mitochondrial genome of A. japonicus are 839 bp in total, with three relatively large regions of Unassigned Sequences (UAS) greater than 100 bp. The gene order of A. japonicus is identical to that observed in the five studied sea urchins, which confirms that the gene order shared by the two classes (Holothuroidea and Echinoidea) is a ground pattern of echinoderm mitochondrial genomes. Bayesian tree based on the cob gene supports the following relationship: (outgroup, (Crinoids, (Asteroids, Ophiuroids, (Echinoids, Holothuroids)))). PMID- 19306916 TI - Cell cycle dependent histone dynamics of an episomal non-viral vector. AB - Non-viral episomal vectors are regarded as attractive alternatives to currently used virus-based vectors in gene therapy. In addition, they represent a minimal model system to study the epigenetic control of basic nuclear processes, such as transcription, replication and nuclear retention. Here we analyze the dynamics of histone modifications during the cell cycle of the episomally replicating vector pEPI-eGFP. The histone code of pEPI-eGFP was compared to its integrating counterpart pGFP-C1. We found that pEPI-eGFP is preferentially associated with histone modifications typical for active chromatin, while pGFP-C1 is mostly decorated with repressive histone modifications. During interphase the distribution of histone modification on pEPI-eGFP is very non-dynamic; the S/MAR shows the highest concentration of active histone modifications. However, they are specifically removed during mitosis and this may correlate with the association and co-segregation of pEPI with the host chromosomes during cell division. PMID- 19306917 TI - Isolation and characterization of DMRT1 and its putative regulatory region in the protogynous wrasse, Halichoeres tenuispinis. AB - A full-length cDNA of doublesex and mab-3-related transcription factor 1 gene (DMRT1) from wrasse testis was isolated by cDNA library screening. Wrasse DMRT1 was 3116 bp in size and contained the DM domain, with a zinc finger DNA-binding motif, and the male-specific motif. Northern blot analysis identified a 3.2-kb transcript approximately equal in size to the DMRT1 nucleotide sequence detected in the testis, but not in the ovary, confirming that this sequence is male specific in protogynous wrasse. Southern blot analysis suggested that the wrasse genome contains two copies of the DMRT1 gene. The ORF consisted of five exons and four introns with conserved donor-acceptor splice sites at all exon-intron junctions. The 5'-flanking region of the wrasse DMRT1 gene was isolated by DNA walking, and putative regulatory sites were identified by searching data bases. The 5'-flanking region was divided into 9 elements, then 17 DMRT-luciferase chimeric plasmids were constructed. By transient transfection into Cos-1 and TM4 cells, distal element I which contains GATA-binding sites and proximal element B containing the sex-determining region on Y chromosome gene (SRY) binding site were revealed to have an important role in transcriptional regulation of the wrasse DMRT1 when an enhancer sequence was provided. PMID- 19306918 TI - Multifunctionality dominantly determines the rate of human housekeeping and tissue specific interacting protein evolution. AB - Elucidation of the determinants of the rate of protein sequence evolution is one of the great challenges in evolutionary biology. It has been proposed that housekeeping genes are evolutionarily slower than tissue specific genes. In the present communication, we have examined different determinants that influence the evolutionary rate variation in human housekeeping and tissue specific proteins present in protein-protein interaction network. Studies on yeast proteome, revealed a predominant role of protein connectivity in determining the rate of protein evolution. However, in human, we did not observe any significant influence of protein connectivity on its evolutionary rate. Rather, a significant impact of the proportion of protein's interacting length (amount of protein interface involved in interaction with its partners), expression level and multifunctionality has been observed in determining the rate of protein evolution. We also observed that multi interface proteins are evolutionarily conserved between housekeeping and tissue specific genes and it has been found that the average number of biological processes they associated in these two sets of genes is similar. Moreover, single interface proteins in housekeeping genes evolve more slowly as compared to tissue specific genes owing to their involvement in different number of biological processes. Partial correlation analysis suggests that the relative importance of three individual factors in determining the evolutionary rate variation between housekeeping and tissue specific proteins is in the order of protein multifunctionality>protein expression level>interacting protein length. PMID- 19306919 TI - Role of myo-inositol phosphate synthase and sucrose synthase genes in plant seed development. AB - The aim of this review is to highlight the role of myo-inositol phosphate synthase (MIPS), which catalyses the first step in inositol biosynthesis and of sucrose synthase (Sus), an enzyme involved in UDP-glucose formation, the principal nucleoside diphosphate in the sucrose cleavage reaction and in trehalose biosynthesis. These two enzymes are involved in various physiological processes including seed growth and resistance to biotic and abiotic stresses. The study of mutated MIPS and Sus genes in some crops, such as soybean and cotton, has shown that these two proteins are directly involved in embryogenesis. They exhibit several isoforms that are essential for normal seed development. The possible role of both genes in seed development is discussed in this review. PMID- 19306920 TI - Differential effects of sodium salicylate on current-evoked firing of pyramidal neurons and fast-spiking interneurons in slices of rat auditory cortex. AB - Sodium salicylate (SS) can penetrate the blood-brain barrier to target neurons in the central auditory system. Understanding how SS alters functional behaviors of different types of central auditory neurons will provide insights into the neural mechanisms of SS-induced tinnitus. Here, we report the differential effects of SS on current-evoked firing of pyramidal neurons and fast-spiking interneurons in layer II/III of auditory cortex slices in young rats (P12-P19). The two neuronal types were identified according to their characteristic patterns of current evoked firing as recorded with whole-cell patch-clamp techniques and by their morphological features. Following perfusion of the brain slice with 1.4mM SS, the threshold current needed to evoke an action potential remained unchanged for pyramidal neurons (68.96+/-10.68 pA vs 70.39+/-12.14 pA, n=7, P>0.05), but significantly increased for fast-spiking interneurons (56.9+/-13.69 pA vs 74.04+/ 15.73 pA, n=7, P<0.05). The drug perfusion caused no significant change in current-evoked firing rates in pyramidal neurons (-2.43+/-7.07%, n=14, P>0.05); however, it drastically and reversibly depressed those in fast-spiking interneurons by up to -49.88+/-10.39% (n=14, P<0.05). Our results suggest that functionally impairing fast-spiking interneurons, which are GABAergic and inhibitory, is probably one of the pathways through which SS raises excitability in the central auditory system and consequently produces tinnitus. PMID- 19306921 TI - An objective method to measure electrode independence in cochlear implant patients with a dual-masker forward masking technique. AB - This study introduced a dual-masker forward masking technique and evaluated whether this objective method could measure electrode independency in a cochlear implant; more particularly, whether the optimal locations and number of active electrodes could be determined. This method further enabled the investigation of the efficacy of current steering, because the proposed recording method could also be described as applying a sequentially current steered masker. The paradigm requires 5 frames involving 2 maskers and 1 probe and is referred to as the Apple Core method (MP5-AC). For each recording, both the masker and probe amplitude were varied independently, producing 3-D eCAP plots that showed the eCAP amplitude for independent variations of masker and probe amplitudes. A simple quantitative model was developed to aid interpretation of the results. Theory and model were clinically tested in 14 patients. On the basis of the model, the multi variate, color-coded plots could be subdivided into seven distinct regions, each depicting a unique relationship between the probe and the maskers. The model's predictions supported interpretation of the results, and indicated independence for the probe electrode contacts only at lower current levels and/or at greater inter-electrode separations. The clinical results revealed a lack of selectivity in the electrode array for stimulus levels larger than 600 microA. This suggests that sequential current steering is only capable of producing a single excitation area at higher current levels, or smaller electrode distances, without additional loudness correction being applied. Thus, the MP5-AC paradigm provided insight concerning the independence of electrodes and the efficacy of current steering in clinical patients. However, its current clinical applicability is limited because measurements were adequate only in anesthetized patients. PMID- 19306922 TI - Free iron ions decrease indoleamine 2,3-dioxygenase expression and reduce IFNgamma-induced inhibition of Chlamydia trachomatis infection. AB - Interferon-gamma (IFNgamma)-mediated indoleamine 2,3-dioxygenase (IDO) expression, important in innate immunity, immune suppression, and tolerance, can be counteracted by ferrous iron (FeSO(4)). Elevation of intracellular iron levels during stimulation with IFNgamma impeded IFNgamma-induced IDO mRNA and protein expression in HEp-2 cells. Decreased IDO expression was accompanied by decreased tryptophan degradation. Accordingly, IFNgamma-mediated suppressing effects on Chlamydia trachomatis (CT) infection were reduced or even abolished in the presence of FeSO(4). Conversely, lowering intracellular iron levels by deferoxamine (DFO) did not increase IFNgamma-induced IDO expression but potentiated Chlamydia-suppressing effects by lowering intracellular iron availability. Additionally, DFO led to a CT-induced IDO expression in HEp-2 cells not treated with IFNgamma. In summary, this study demonstrates that iron acts as a regulatory element for modulating IDO expression, in addition to its function as an essential element for chlamydial growth. This may represent an important control mechanism of IDO expression at the transcriptional level. PMID- 19306923 TI - Comparative in vitro toxicity of seven zinc-salts towards neuronal PC12 cells. AB - Currently much attention has been given to the neurotoxicity of zinc, yet little is known about the influence of the counterions present. Therefore, we investigated the influence of different Zn(2+)-salts (concentrations range 0.05 0.3 mM) on cell viability, ATP and glutathione concentration and caspase activation in differentiated PC12 cells as a model for neuronal cells. Generally, at concentrations of 0.05 mM most Zn(2+)-salts were not cytotoxic except for zinc citrate. At concentrations between 0.1 and 0.3 mM Zn(2+) a significant decrease in GSH and ATP levels preceded cell death induced by all salts, except of zinc histidinate. Zinc-citrate and zinc-sulphate turned out to be the most toxic salts particularly at low concentrations. Analyses of caspase 3/7 activity showed that dependent on the concentration and the type of the salt used cell death may show more or less signs of both, necrosis and apoptosis. Interestingly, the uptake of Zn(2+) from zinc-sulphate and zinc-citrate was significantly higher than that of other salts, implicating a correlation between uptake and toxicity. In conclusion, Zn(2+)-salts could be divided into three categories with high (zinc citrate, zinc-sulphate), moderate (zinc-orotate, zinc-acetate, zinc-chloride(,) zinc-gluconate) and low cytotoxicity (zinc-histidinate). PMID- 19306924 TI - Pathway prediction by bioinformatic analysis of the untranslated regions of the CFTR mRNA. AB - Mining the information contained within the genetic code in untranslated regions has proven difficult because of the ambiguity of microRNA and protein binding sites. This manuscript describes a bioinformatic screen that identifies long sequences with partial identity to the untranslated regions of the cystic fibrosis transmembrane regulator. This screen uncovered a long, evolutionarily conserved motif common to the 3' UTRs of the CFTR and SEC24A transcripts, and shorter, statistically significant motifs unique to either 5' or 3' UTRs. In addition, of the 140 transcripts identified in the screen that encode proteins with known protein interactions, 130 are linked to CFTR through protein interactions. The screen identified genes that are known to be involved in lung fibrosis, the inflammatory response of cystic fibrosis and sensitivity to Pseudomonas aeruginosa infections. The bioinformatic analysis of untranslated regions should prove to be a powerful adjunct to other tools for predicting pathways and relevant interactions. PMID- 19306925 TI - Dual regulation of lysophosphatidic acid (LPA1) receptor signalling by Ral and GRK. AB - Lysophosphatidic acid (LPA) is a major constituent of blood and is involved in a variety of physiological and pathophysiological processes. LPA signals via the ubiquitously expressed G protein-coupled receptors (GPCRs), LPA(1) and LPA(2) that are specific for LPA. However, in large, the molecular mechanisms that regulate the signalling of these receptors are unknown. We show that the small GTPase RalA associates with both LPA(1) and LPA(2) in human embryonic kidney (HEK 293) cells and that stimulation of LPA(1) receptors with LPA triggers the activation of RalA. While RalA was not found to play a role in the endocytosis of LPA receptors, we reveal that LPA(1) receptor stimulation promoted Ral-dependent phospholipase C activity. Furthermore, we found that GRK2 is required for the desensitization of LPA(1) and LPA(2) and have identified a novel interaction between RalA and GRK2, which is promoted by LPA(1) receptor activity. Taken together, these results establish RalA and GRK2 as key regulators of LPA receptor signalling and demonstrate for the first time that LPA(1) activity facilitates the formation of a novel protein complex between these two proteins. PMID- 19306926 TI - beta-Arrestins facilitate ubiquitin-dependent degradation of apoptosis signal regulating kinase 1 (ASK1) and attenuate H2O2-induced apoptosis. AB - beta-Arrestins are ubiquitously expressed proteins that play important roles in receptor desensitization, endocytosis, proteosomal degradation, apoptosis and signaling. It has been reported that beta-Arrestin2 acts as a scaffold by directly interacting with the JNK3 isoform and recruiting MKK4 and the apoptosis signaling kinase-1 (ASK1). Here, we report a novel function of beta-Arrestins in regulating H(2)O(2)-induced apoptosis. Our study demonstrates that beta-Arrestins physically associate with C-terminal domain of ASK1, and moreover, both over expression and RNA interference (RNAi) experiments indicate that beta-Arrestins down-regulate ASK1 protein. In detail, beta-Arrestin-induced reduction of ASK1 protein is due to ubiquitination and proteasome-dependent degradation of ASK1 in response to association of beta-Arrestins and ASK1. Upon H(2)O(2) stimulation, the protein binding between beta-Arrestins and ASK1 increases and ASK1 degradation is expedited. In consequence, beta-Arrestins prevent ASK1-JNK signaling and as a result attenuate H(2)O(2)-induced apoptosis. Structurally, C terminal domain of ASK1 is essential for beta-Arrestins and ASK1 association. We also found that CHIP is required for beta-Arrestins-induced ASK1 degradation, which suggested that beta-Arrestins function as a scaffold of ASK1 and CHIP, leading to CHIP-mediated ASK1 degradation. All these findings indicate that beta Arrestins play a negative regulatory role in H(2)O(2)-induced apoptosis signaling through associating with ASK1 and CHIP and facilitating ASK1 degradation, which provides a new insight for analyzing the effects of beta-Arrestins on protecting cells from oxidative stress-induced apoptosis. PMID- 19306927 TI - Annexin-A6 presents two modes of association with phospholipid membranes. A combined QCM-D, AFM and cryo-TEM study. AB - Annexins are soluble proteins that bind to biological membranes in a Ca(2+) dependent manner. Annexin-A6 (AnxA6) is unique in the annexin family as it consists of the repeat of two annexin core modules, while all other annexins consist of a single module. AnxA6 has been proposed to participate in various membrane-related processes, including endocytosis and exocytosis, yet the molecular mechanism of association of AnxA6 with biological membranes, especially its ability to aggregate membranes, is still unclear. To address this question, we studied the association of AnxA6 with model phospholipid membranes by combining the techniques of quartz crystal microbalance with dissipation monitoring (QCM-D), (cryo-) transmission electron microscopy (TEM) and atomic force microscopy (AFM). The properties of membrane binding and membrane aggregation of AnxA6 were compared to two reference systems, annexin A5 (AnxA5), which is the annexin prototype, and a chimerical AnxA5-dimer molecule, which is able to aggregate two membranes in a symmetrical manner. We show that AnxA6 presents two modes of association with lipid membranes depending on Ca(2+) concentration. At low Ca(2+)-concentration ( approximately 60-150microM), AnxA6 binds to membranes via its two coplanar annexin modules and is not able to associate two separate membranes. At high Ca(2+)-concentration ( approximately 2mM), AnxA6 molecules are able to bind two adjacent phospholipid membranes and present a conformation similar to the AnxA6 3D crystallographic structure. Possible biological implications of these novel membrane-binding properties of AnxA6 are discussed. PMID- 19306928 TI - In situ molecular level studies on membrane related peptides and proteins in real time using sum frequency generation vibrational spectroscopy. AB - Sum frequency generation (SFG) vibrational spectroscopy has been demonstrated to be a powerful technique to study the molecular structures of surfaces and interfaces in different chemical environments. This review summarizes recent SFG studies on hybrid bilayer membranes and substrate-supported lipid monolayers and bilayers, the interaction between peptides/proteins and lipid monolayers/bilayers, and bilayer perturbation induced by peptides/proteins. To demonstrate the ability of SFG to determine the orientations of various secondary structures, studies on the interactions between different peptides/proteins (melittin, G proteins, alamethicin, and tachyplesin I) and lipid bilayers are discussed. Molecular level details revealed by SFG in these studies show that SFG can provide a unique understanding on the interactions between a lipid monolayer/bilayer and peptides/proteins in real time, in situ and without any exogenous labeling. PMID- 19306929 TI - Task complexity relates to activation of cortical motor areas during uni- and bimanual performance: a functional NIRS study. AB - Hand motor tasks are frequently used to assess impaired motor function in neurology and neurorehabilitation. Assessments can be varied by means of hand laterality, i.e. unimanual or bimanual performance, as well as by means of task complexity, i.e. different degrees ranging from simple to complex sequence tasks. The resulting functional activation in human primary motor cortex (M1) has been studied intensively by traditional neuroimaging methods. Previous studies using functional near-infrared spectroscopy (fNIRS) investigated simple hand motor tasks. However, it is unknown whether fNIRS can also detect changes in response to increasing task complexity. Our hypothesis was to show that fNIRS could detect activation changes in relation to task complexity in uni- and bimanual tasks. Sixteen healthy right-handed subjects performed five finger-tapping tasks: unimanual left and right, simple and complex tasks as well as bimanual complex tasks. We found significant differences in oxy-hemoglobin (O(2)Hb) and deoxy hemoglobin (HHb) concentration in the right hemisphere over M1. Largest O(2)Hb concentration changes were found during complex (0.351+/-0.051 micromol/l) and simple (0.275+/-0.054 micromol/l) right hand tasks followed by bimanual (0.249+/ 0.047 micromol/l), complex (0.154+/-0.034 micromol/l) and simple (0.110+/-0.034 micromol/l) left hand tasks. Largest HHb concentration changes were found during bimanual (-0.138+/-0.006 micromol/l) tasks, followed by simple right hand ( 0.12+/-0.016 micromol/l), complex left (-0.0875+/-0.007 micromol/l), complex right (-0.0863+/-0.005 micromol/l) and simple left (-0.0674+/-0.005 micromol/l) hand tasks. We report for the first time that fNIRS detects oxygenation changes in relation to task complexity during finger-tapping. The study aims to contribute to the establishment of fNIRS as a neuroimaging method to assess hand motor function in clinical settings where traditional neuroimaging methods cannot be applied. PMID- 19306930 TI - Reward circuitry is perturbed in the absence of the serotonin transporter. AB - The serotonin transporter (SERT) modulates the entire serotonergic system in the brain and influences both the dopaminergic and norepinephrinergic systems. These three systems are intimately involved in normal physiological functioning of the brain and implicated in numerous pathological conditions. Here we use high resolution magnetic resonance imaging (MRI) and spectroscopy to elucidate the effects of disruption of the serotonin transporter in an animal model system: the SERT knock-out mouse. Employing manganese-enhanced MRI, we injected Mn(2+) into the prefrontal cortex and obtained 3D MR images at specific time points in cohorts of SERT and normal mice. Statistical analysis of co-registered datasets demonstrated that active circuitry originating in the prefrontal cortex in the SERT knock-out is dramatically altered, with a bias towards more posterior areas (substantia nigra, ventral tegmental area, and Raphe nuclei) directly involved in the reward circuit. Injection site and tracing were confirmed with traditional track tracers by optical microscopy. In contrast, metabolite levels were essentially normal in the SERT knock-out by in vivo magnetic resonance spectroscopy and little or no anatomical differences between SERT knock-out and normal mice were detected by MRI. These findings point to modulation of the limbic cortical-ventral striatopallidal by disruption of SERT function. Thus, molecular disruptions of SERT that produce behavioral changes also alter the functional anatomy of the reward circuitry in which all the monoamine systems are involved. PMID- 19306931 TI - A MEG investigation of somatosensory processing in the rhesus monkey. AB - The use of minimally and non-invasive neuroimaging methods in animal models has sharply increased over the past decade. Such studies have enhanced understanding of the neural basis of the physical signals quantified by these tools, and have addressed an assortment of fundamental and otherwise intractable questions in neurobiology. To date, these studies have almost exclusively utilized positron emission tomography or variants of magnetic resonance based imaging. These methods provide largely indirect measures of brain activity and are strongly reliant on intact vasculature and normal blood-flow, which is known to be compromised in many clinical conditions. The current study provides the first demonstration of whole-head magnetoencephalography (MEG), a non-invasive and direct measure of neuronal activity, in a rhesus monkey, and in the process supplies the initial data on systems-level dynamics in somatosensory cortices. An adult rhesus monkey underwent three separate studies of tactile stimulation on the pad of the right second or fifth digit as whole-head MEG data were acquired. The neural generators of the primary neuromagnetic components were localized using an equivalent-current-dipole model. Second digit stimulation produced an initial cortical response peaking approximately 16 ms after stimulus onset in the contralateral somatosensory cortices, with a later response at approximately 96 ms in an overlapping or nearby neural area with a roughly orthogonal orientation. Stimulation of the fifth digit produced similar results, the main exception being a substantially weaker later response. We believe the 16 ms response is likely the monkey homologue of the human M50 response, as both are the earliest cortical response and localize to the contralateral primary somatosensory area. Thus, these data suggest that mechanoreception in nonhuman primates operates substantially faster than that in adult humans. More broadly, these results demonstrate that it is feasible to use current human whole-head MEG instrumentation to record neuromagnetic responses in adult rhesus monkeys. Nonhuman primate models of human disease provide the closest phylogenetic link to humans. The present, non-invasive imaging study could promote exciting translational integration of invasive animal studies and non-invasive human studies, allowing experimentally induced deficits and pharmacological treatments to be interpreted in light of resulting brain network interactions. PMID- 19306932 TI - Bayesian model selection for group studies. AB - Bayesian model selection (BMS) is a powerful method for determining the most likely among a set of competing hypotheses about the mechanisms that generated observed data. BMS has recently found widespread application in neuroimaging, particularly in the context of dynamic causal modelling (DCM). However, so far, combining BMS results from several subjects has relied on simple (fixed effects) metrics, e.g. the group Bayes factor (GBF), that do not account for group heterogeneity or outliers. In this paper, we compare the GBF with two random effects methods for BMS at the between-subject or group level. These methods provide inference on model-space using a classical and Bayesian perspective respectively. First, a classical (frequentist) approach uses the log model evidence as a subject-specific summary statistic. This enables one to use analysis of variance to test for differences in log-evidences over models, relative to inter-subject differences. We then consider the same problem in Bayesian terms and describe a novel hierarchical model, which is optimised to furnish a probability density on the models themselves. This new variational Bayes method rests on treating the model as a random variable and estimating the parameters of a Dirichlet distribution which describes the probabilities for all models considered. These probabilities then define a multinomial distribution over model space, allowing one to compute how likely it is that a specific model generated the data of a randomly chosen subject as well as the exceedance probability of one model being more likely than any other model. Using empirical and synthetic data, we show that optimising a conditional density of the model probabilities, given the log-evidences for each model over subjects, is more informative and appropriate than both the GBF and frequentist tests of the log evidences. In particular, we found that the hierarchical Bayesian approach is considerably more robust than either of the other approaches in the presence of outliers. We expect that this new random effects method will prove useful for a wide range of group studies, not only in the context of DCM, but also for other modelling endeavours, e.g. comparing different source reconstruction methods for EEG/MEG or selecting among competing computational models of learning and decision-making. PMID- 19306933 TI - Mapping the bilateral visual integration by EEG and fMRI. AB - In the human visual system, the internal representation of the left and right visual hemifields is split at the midline of the two cerebral hemispheres. The present study aims to address the questions of when and where the lateralized cortical visual representations are merged to form an intact percept by using a multimodal neuroimaging approach. Visual evoked potential (VEP) and functional magnetic resonance imaging (fMRI) data were acquired from a group of healthy subjects presented with unilateral versus bilateral visual stimuli. Cortical activities involved in processing bilateral visual information are expected to be equally responsive to ipsilateral and contralateral stimuli, and demonstrate spatial nonlinearity in the response to bilateral stimuli. Utilizing these features, we performed integrative as well as separate analyses for both VEP and fMRI data. The present results suggest that i) the majority of cortical activity that integrates visual information across hemifields takes place at extrastriate areas during late visual processing, and that ii) the lateral occipito-temporal (LOT) regions (likely the MT+ complex) and the medial occipital cortex (i.e. V1) may contribute to bilateral visual integration during early visual processing. Our findings are generally in agreement with the bottom-up visual hierarchy, with the exception of the evidence suggesting an early activation of the higher-tier LOT areas and the influence from ipsilateral visual inputs upon the V1 response. PMID- 19306934 TI - Porous genomes and species integrity in the branchiopod Artemia. AB - Over the years, studies on interspecific hybridization have highlighted cases where gene exchange between taxa continues for a significant amount of time after speciation. The reasons for this lag of reproductive isolation relative to genetic isolation are largely unclear, and the question still remains whether the resulting hybrids represent novel biological (and taxonomic) diversity or merely an evolutionary liability. We provide strong indications in the branchiopod Artemia that hybrids between distantly related species may not be evolutionary inconsequential. Based on a global sampling of published and newly derived nuclear (ITS1) and mitochondrial (16S rRNA) sequence data from all representatives of the genus, we have identified natural hybrids between Artemia species (A. persimilisxA. franciscana, A. salinaxA. franciscana) separated by evolutionary interludes of tens of millions of years. Our combined analytical framework of cladistic and network methods provides evidence that hybridizations are the result of recent secondary contact following pronounced allopatric differentiation. The detection of mitochondrial introgression from A. persimilis to A. franciscana attests F(1) hybrid fertility. The reasons for this apparent unidirectionality of introgression are currently unknown but a likely explanation is provided based on morphometric divergence. We discuss the evolutionary implications of our results within the broader context of continental zooplankters. PMID- 19306935 TI - Phylogenetic analyses of complete mitochondrial genome of Urechis unicinctus (Echiura) support that echiurans are derived annelids. PMID- 19306936 TI - The polyploid series of Centaurea toletana: glacial migrations and introgression revealed by nrDNA and cpDNA sequence analyzes. AB - The polyploid series of Centaurea toletana comprises diploid, tetraploid, and hexaploid cytotypes. Previous studies suggested that the tetraploid was an autopolyploid, while the hexaploid was an allopolyploid and should be considered a different species, C. argecillensis. Sequencing of the ITS and rps4-trnT-trnL, ycf3-trnS, and rpL16 regions, and extensive cloning and sequencing of the ETS region have revealed that many diploid individuals and populations show different ribotypes, likely resulting from ancient hybridization events. Ribotypes found in the diploid populations are also present in tetraploid populations. The extreme difficulties in classifying the tetraploid as auto- or allopolyploid are discussed. The hexaploid C. argecillensis also shows many different ribotypes, including a ribotype not found in the diploids and making an autopolyploid origin unlikely. The pattern of introgression and gene flow implicates several species from the Iberian Peninsula and the High Atlas Mountains in Morocco as genetic donors in ancient hybridization events. This long-reaching network of hybridization may trace its origin to the climatic history of the western Mediterranean during the Neogene. PMID- 19306937 TI - Molecular phylogeny of parabasalids with emphasis on the order Cristamonadida and its complex morphological evolution. AB - Parabasalia represents a complex assemblage of species, which recently received extensive reorganization. The newly created order Cristamonadida unites complex hypermastigids belonging to the Lophomonadida like the joeniids, the multinucleate polymonad Calonymphidae, and well-developed trichomonads in the Devescovinidae. All these protists exclusively occur in the guts of termites and related insects. In this study, small subunit rRNA and glyceraldehyde-3-phosphate dehydrogenase genes were identified without cultivation from 14 species in Cristamonadida including previously unstudied genera such as Joenina, Joenia, Joenoides, Macrotrichomonas, Gigantomonas, and Foaina. Despite the great morphological diversity of Cristamonadida, our phylogenetic analyses supported the monophyly of this order. However, almost all the families and subfamilies composing this order are polyphyletic suggesting a complicated morphological evolution. Our analyses also showed that Cristamonadida descends from one lineage of rudimentary trichomonads and that joeniids was basal in this order. Several successive and independent morphological transitions such as the development and reduction of flagellar apparatus and associated cytoskeleton and transition to multinucleated status have likely led to the diversity and complexity of cristamonad lineages. PMID- 19306938 TI - Downregulation of CD2-associated protein impaired the physiological functions of podocytes. AB - Emerging evidences show that CD2-associated protein (CD2AP) is involved in podocyte injury and the pathogenesis of proteinuria. However, the exact molecular mechanism by which CD2AP exerts its biological function is elusive. We knocked down CD2AP gene by target siRNA in conditionally immortalized mouse podocytes, which showed lowered cell adhesion and spreading ability (P<0.05). At the same time, cell cycle was arrested in G2/M phase (P<0.05), and pathologic nuclear division could easily be seen in CD2AP siRNA-transfected podocytes. The proliferation of podocytes were also inhibited significantly by CD2AP siRNA transfection (P<0.05). Further study revealed disordered distributions of F actin, as well as lowered nephrin expression and phosphorylation in podocytes. These data suggest that CD2AP may play a crucial role in maintaining the normal function of podocytes and lowered CD2AP causes podocyte injury by disrupting the cytoskeleton and disturbing the nephrin-CD2AP signaling pathway. PMID- 19306939 TI - Genomic structure, expression pattern and functional characterization of crustinPm5, a unique isoform of crustin from Penaeus monodon. AB - A unique isoform of crustin, crustinPm5, was identified from a gill-epipodite cDNA library of the tiger shrimp, Penaeus monodon. The crustinPm5 cDNA contains an open reading frame (ORF) of 510 bp encoding a 169 amino acid protein. The deduced amino acid sequence of crustinPm5 showed 38% and 37% overall sequence identity with those of crustinPm1 and crustin-likePm, respectively, two crustin isoforms previously reported. The crustinPm5 gene contained four exons interrupted by three introns whilst the upstream sequence contains a putative promoter with two potential binding sites for NF-kappaB, one complete heat-shock regulatory element (HSE) and five putative GATA factor binding sites. The transcripts of crustinPm5 were primarily observed in the epipodite and eyestalk and not in hemocytes. Expression analysis revealed that the transcript levels of crustinPm5, crustinPm1 and crustin-likePm in epipodite were up-regulated upon heat treatment and hyperosmotic salinity stress. The recombinant crustinPm5 exhibited antimicrobial activity against some Gram-positive bacteria in vitro, but did not inhibit the growth of any Gram-negative bacteria tested. These results, together with the transcript expression pattern, indicate a diverse function of the proteins in the crustin family particularly crustinPm5 that might function as a stress mediator in addition to its antibacterial action. PMID- 19306940 TI - The incidence of Streptococcus pneumoniae otitis media is affected by the polymicrobial environment particularly Moraxella catarrhalis in a mouse nasal colonisation model. AB - Otitis media (OM) is a highly prevalent paediatric disease with both bacterial and viral triggers of infection. This study has investigated how combinations of bacteria associated with nasal colonisation and the occurrence and absence of viral infection (Sendai virus) induce OM in a mouse nasal colonisation model. The respiratory virus significantly contributed to bacterial OM for all bacterial combinations (p<0.001). Streptococcus pneumoniae consistently dominated as the causative bacterium of OM and when co-infected with S. pneumoniae, Moraxella catarrhalis more significantly affected pneumococcal OM than did non-typeable Haemophilus influenzae (p<0.001) by increasing the incidence rate, infection bacterial load and duration of infection. Nitric oxide levels in the middle ear, an indicator of inflammation, peaked at day 3 in single bacterium groups, but at day 1 in mixed bacterial groups and was produced in all bacteria inoculated groups even in the absence of viable bacterial recovery. Phagocytic cells were recruited rapidly to the ear following nasal inoculation but over time their numbers did not correlate with persistence of bacterial infection. The study has shown that the composition of bacteria in the nasal cavity and respiratory viral infection significantly affected the OM incidence rate, duration of infection and bacterial load (severity). PMID- 19306941 TI - Proton pump inhibitors for gastroduodenal damage related to nonsteroidal anti inflammatory drugs or aspirin: twelve important questions for clinical practice. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) and aspirin are among the most commonly used medications worldwide. Their use is associated with significant gastroduodenal adverse effects, including dyspepsia, bleeding, ulcer formation, and perforation. Given their long-term use by millions of patients, there is a substantial impact at the population level of these complications. In this evidence-based review, we have endeavored to answer 12 commonly encountered questions in clinical practice that deal with the following: extent of the problem of NSAID/aspirin-induced gastroduodenal damage and its impact on public health; role of proton pump inhibitors (PPIs) in the primary prevention, healing, and secondary prevention of NSAID/aspirin-induced gastroduodenal ulceration as assessed by using endoscopic end points; role of PPIs in the prevention of adverse clinical outcomes related to NSAID/aspirin use; whether PPIs are effective in NSAID-induced dyspepsia; comparison of PPI co-therapy with selective cyclooxygenase-2 inhibitors for risk reduction of adverse clinical outcomes; role of PPIs in preventing rebleeding from aspirin +/- clopidogrel therapy in high risk patients; identifying high-risk patients who can benefit from PPI co therapy; the role of other gastroprotective agents for prevention of NSAID/aspirin-induced gastroduodenal damage; and the cost-effectiveness of and limitations to the use of PPIs for prevention of gastroduodenal damage related to the use of NSAIDs or aspirin. We then summarized our recommendations on the use of PPIs for the clinical management of patients using NSAIDs or aspirin. PMID- 19306942 TI - Image of the month. Intestinal spirochetosis: an unusual cause of asymptomatic colonic ulceration. PMID- 19306943 TI - Endoscopic therapy for Barrett's esophagus. AB - Recent retrospective cohort data and a prospective randomized sham controlled trial have clearly documented the impact of endoscopic ablation therapy on dysplasia and Barrett's esophagus (BE). The clinical indications for ablation of BE includes high-grade dysplasia and intramucosal adenocarcinoma. The techniques of resection of mucosal irregularities and of ablation are reviewed, primarily thermal and photodynamic ablation. Ablation of BE with neoplasia has appropriately entered the clinical arena. PMID- 19306944 TI - Image of the month. Primary epiploic appendagitis. PMID- 19306945 TI - A comparison between whites and blacks with severe multi-organ iron overload identified in 16,152 autopsies. AB - BACKGROUND & AIMS: Little is known about differences in the prevalence of severe iron overload at death in whites and blacks. We evaluated data and samples from 16,152 autopsies (8484 whites, 7668 blacks) performed at a single university hospital. METHODS: Cases of severe multi-organ iron overload were identified by review of autopsy protocols and Perls-stained tissue specimens, analysis of hepatocyte and Kupffer cell iron levels, and measurement of liver tissue iron concentrations. RESULTS: We analyzed autopsy data from 10,345 adults (age > or =21 years), 1337 children (1-20 years), and 4470 infants (<1 year). Iron overload without reports of excessive exogenous iron was observed in 18 adults; the prevalence in whites and blacks was 0.0019 and 0.0015, respectively (P = .6494). Twenty-nine adults and 2 children had iron overload with reports of excessive exogenous iron. In adults, the prevalences of iron overload with reports of excessive exogenous iron in whites and blacks were 0.0040 and 0.0013, respectively (P = .0107). Among adults, the prevalence of cirrhosis was 6-fold greater in those with iron overload. In adults with severe iron overload, 67% without reports of excessive exogenous iron and 14% with reports of excessive exogenous iron died of hepatic failure or cardiomyopathy caused by siderosis. The overall prevalence of deaths caused by severe iron overload in whites and blacks was 0.0021 and 0.0009, respectively (P = .0842). CONCLUSIONS: The prevalence of severe iron overload without reports of excessive exogenous iron did not differ significantly between whites and blacks. The prevalence of iron overload with reports of excessive exogenous iron was greater in whites. Hepatic failure and cardiomyopathy were common causes of death in severe iron overload cases. PMID- 19306946 TI - Poorly responsive ulcerative colitis in the hospital. PMID- 19306947 TI - Recurrent gastrointestinal bleeding after negative upper endoscopy and colonoscopy. PMID- 19306948 TI - Image of the month. Massive cerebral air embolism during esophagogastroduodenoscopy. PMID- 19306949 TI - Health-related quality of life predicts mortality in patients with advanced chronic liver disease. AB - BACKGROUND & AIMS: It is well-established that cirrhosis negatively impacts health-related quality of life (HRQOL). However, it is less clear how to use this information in everyday clinical practice. If HRQOL predicted survival in cirrhosis, then measuring HRQOL would have important clinical implications. We sought to measure the association between HRQOL and survival in patients with cirrhosis and investigated whether the relationship between HRQOL and survival is independent of Model for End-Stage Liver Disease (MELD). METHODS: We measured HRQOL in 156 patients with cirrhosis awaiting liver transplantation by using the Short Form Liver Disease Quality of Life instrument. We followed patients prospectively and used Cox proportional hazard models to measure the independent effect of baseline HRQOL on survival, adjusting for MELD and other covariates. RESULTS: During a mean 9-month follow-up, 26 (17%) patients died, and 30 (20%) received liver transplants. In unadjusted analysis, higher baseline HRQOL predicted lower mortality (hazard ratio, 0.96; 95% confidence interval, 0.94 0.99). Specifically, for each 1-point increase in HRQOL, there was a 4% decrease in mortality. These results did not change after adjusting for MELD scores, patient demographics, or psychosocial characteristics; the MELD score accounted for 1% of the variation in HRQOL scores (P = .18). Survival was most strongly predicted by activities of daily living, health distress, sleep disturbance, and perceived disease stigma. CONCLUSIONS: Higher HRQOL predicts lower mortality in patients with cirrhosis. This relationship is independent of MELD; MELD does not capture liver-specific HRQOL. Beyond its use as a secondary outcome in clinical trials, HRQOL could be used to predict survival of patients with advanced liver disease. PMID- 19306950 TI - TNF-alpha regulates vascular smooth muscle cell responses in genetic hypertension. AB - Cellular and molecular events in vascular smooth muscle cells (VSMC) from Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were investigated. SHR derived VSMC showed increased proliferative capacity and MAP kinase levels in comparison with WKY-derived VSMC. Flow cytometry analysis revealed that progression from G1 to S phase was faster in SHR-derived VSMC in response to tumor necrosis factor-alpha (TNF-alpha) as compared with cells from WKY. The G1 cell cycle-associated proteins such as cyclin D1, cyclin E, CDK2 and CDK4, and kinase activities associated with CDK2 and CDK4, were increased in SHR-derived VSMC. In addition, CDK inhibitor p21 was elevated in SHR-derived cells. Matrix metalloproteinase-9 (MMP-9) expression and migration were also increased in response to TNF-alpha in SHR-derived cells. This increase was characterized by the up-regulation of MMP-9, which was transcriptionally regulated at the AP-1 and NF-kappaB sites in the MMP-9 promoter. These results suggest that the hypertensive-associated increase in VSMC proliferative capacity, G1 to S-phase cell-cycle progress and MMP-9 expression may play a role in vascular remodeling in hypertension. PMID- 19306951 TI - Clinical significance of antiphosphatidylethanolamine antibodies in the so-called "seronegative antiphospholipid syndrome". AB - The antiphospholipid syndrome (APS) is characterized by thrombosis and/or pregnancy morbidity associated with the presence of laboratory criteria such as antibodies directed towards cardiolipin or beta(2)-GPI and lupus anticoagulant. Recently, the term "seronegative APS " has been proposed to define patients with the typical clinical manifestations but with negative serologies. One explanation for such a context could be that some APS patients may only have antiphospholipid antibodies (aPL) other than the admitted laboratory criteria. This review is focused on antibodies directed against phosphatidylethanolamine (aPE) and underlines the interest of their investigation in the different clinical manifestations of APS. PMID- 19306952 TI - Attenuation of ionic interactions profoundly lowers the kinetic thermal stability of Pyrococcus furiosus triosephosphate isomerase. AB - We investigate here the high structural stability of Pyrococcus furiosus triosephosphate isomerase (PfuTIM) by exploring the effects - upon the protein's structure and kinetic thermal stability - of modulation of its ionic interactions through pH variations, and mutations. PfuTIM shows comparable structural contents at pH 3.0, 7.0 and 10.0. However, at pH 3.0, subtle changes are seen in the protein's surface hydrophobicity and association status, and its kinetic thermal stability is profoundly reduced (as evidenced by its facile heat- and cold mediated denaturation, characterized by a high degree of hysteresis and irreversibility). Increase in ionic strength through addition of salt counters the reduction of stability, and reversal of pH facilitates partial refolding. Further, a mutated form of PfuTIM (mPfuTIM) lacking 4 key charged residues involved in ionic interactions displays a structural content identical to PfuTIM but profound reduction in kinetic stability to thermal and chemical denaturation, as well as evidence of partial unfolding at temperatures between 90 degrees C and 100 degrees C, unlike PfuTIM. We conclude, therefore, that ionic interactions (which are known to determine protein thermodynamic stability) can also contribute significantly to protein kinetic thermal stability. PMID- 19306953 TI - Extending the phenotype of recurrent rearrangements of 16p11.2: deletions in mentally retarded patients without autism and in normal individuals. AB - Array CGH (comparative genomic hybridization) screening of large patient cohorts with mental retardation and/or multiple congenital anomalies (MR/MCA) has led to the identification of a number of new microdeletion and microduplication syndromes. Recently, a recurrent copy number variant (CNV) at chromosome 16p11.2 was reported to occur in up to 1% of autistic patients in three large autism studies. In the screening of 4284 patients with MR/MCA with various array platforms, we detected 22 individuals (14 index patients and 8 family members) with deletions in 16p11.2, which are genomically identical to those identified in the autism studies. Though some patients shared a facial resemblance and a tendency to overweight, there was no evidence for a recognizable phenotype. Autism was not the presenting feature in our series. The assembled evidence indicates that recurrent 16p11.2 deletions are associated with variable clinical outcome, most likely arising from haploinsufficiency of one or more genes. The phenotypical spectrum ranges from MR and/or MCA, autism, learning and speech problems, to a normal phenotype. PMID- 19306954 TI - Increased circulating heat shock protein 60 induced by menopause, stimulates apoptosis of osteoblast-lineage cells via up-regulation of toll-like receptors. AB - Postmenopausal osteoporosis is a heterogeneous disorder characterized by accelerated bone loss after natural or surgical menopause and an increased risk of fractures. The bone loss in estrogen deficiency results from the increased bone resorption and impaired ability of osteoblastic bone formation. Previous studies have reported that the HSP60 stimulates osteoclast formation and bone resorption. Here we found that plasma HSP60 levels were significantly higher in postmenopausal (median 1152.4 ng/ml; range 724.7-2123.4 ng/ml) than in premenopausal (median 316.3 ng/ml; range 164.6-638.4 ng/ml) women. In primary human bone marrow stromal cells (hBMSC) and the HS-5 hBMSC cell line, HSP60 significantly reduced cell viability and increased caspase-dependent apoptosis. Consistent with these observations, HSP60 activated caspase-3 and -9, but not caspase-8 in HS-5 cells, and increased the release of mitochondrial cytochrome c into the cytosol. In addition, HSP60 activated p38 and NFkappaB, but not ERK or JNK; importantly, inhibitors of p38 (SB203580) and NFkappaB (PDTC) abolished HSP60-induced apoptosis. Furthermore, Western blotting showed that HSP60 up regulated TLR-2 and TLR-4 expression, and pretreatment with blocking antibodies for TLR-2 and TLR-4 almost completely eliminated the effects of HSP60 on apoptosis, caspase-3 and -9 activation, and activation of NFkappaB and p38 MAPK. Most notably, ovariectomy-induced bone loss was attenuated in TLR-2 KO mice. In conclusion, up-regulation of TLR-2 by HSP60 may play a critical role in promoting bone loss in the estrogen-deficient state. PMID- 19306955 TI - Early diet and peak bone mass: 20 year follow-up of a randomized trial of early diet in infants born preterm. AB - BACKGROUND: Preterm infants are at risk of metabolic bone disease due to inadequate mineral intake with unknown consequences for later bone health. OBJECTIVE: To test the hypotheses that (1) early diet programs peak bone mass and bone turnover; (2) human milk has a beneficial effect on these outcomes; (3) preterm subjects have reduced peak bone mass compared to population reference data. DESIGN: 20 year follow-up of 202 subjects (43% male; 24% of survivors) who were born preterm and randomized to: (i) preterm formula versus banked breast milk or (ii) preterm versus term formula; as sole diet or supplement to maternal milk. Outcome measures were (i) anthropometry; (ii) hip, lumbar spine (LS) and whole body (WB) bone mineral content (BMC) and bone area (BA) measured using DXA; (iii) bone turnover markers. RESULTS: Infant dietary randomization group did not influence peak bone mass or turnover. The proportion of human milk in the diet was significantly positively associated with WBBA and BMC. Subjects receiving >90% human milk had significantly higher WBBA (by 3.5%, p=0.01) and BMC (by 4.8%, p=0.03) than those receiving <10%. Compared to population data, subjects had significantly lower height SDS (-0.41 (SD 1.05)), higher BMI SDS (0.31 (1.33)) and lower LSBMD SDS (-0.29 (1.16)); height and bone mass deficits were greatest in those born SGA with birthweight <1250 g (height SDS -0.81 (0.95), LSBMD SDS 0.61 (1.3)). CONCLUSION: Infant dietary randomization group did not affect peak bone mass or turnover suggesting the observed reduced final height and LS bone mass, most marked in growth restricted subjects with the lowest birthweight, may not be related to sub-optimal early nutrition. The higher WB bone mass associated with human milk intake, despite its low nutrient content, may reflect non nutritive factors in breast milk. These findings may have implications for later osteoporosis risk and require further investigation. PMID- 19306956 TI - New insights into BMP-7 mediated osteoblastic differentiation of primary human mesenchymal stem cells. AB - Bone Morphogenetic Proteins (BMPs) are members of the TGF-beta superfamily of growth factors. Several BMPs exhibit osteoinductive bioactivities, and are critical for bone formation in both developing and mature skeletal systems. BMP-7 (OP-1) is currently used clinically in revision of posterolateral spine fusions and long bone non-unions. The current study characterizes BMP-7 induced gene expression during early osteoblastic differentiation of human mesenchymal stem cells (hMSC). Primary hMSC were treated with BMP-7 for 24 or 120 h and gene expression across the entire human genome was evaluated using Affymetrix HG-U133 Plus 2.0 Arrays. 955 probe sets representing 655 genes and 95 ESTs were identified as differentially expressed and were organized into three major expression profiles (Profiles A, B and C) by hierarchical clustering. Genes from each profile were classified according to biochemical pathway analyses. Profile A, representing genes upregulated by BMP-7, revealed strong enrichment for established osteogenic marker genes, as well as several genes with undefined roles in osteoblast function, including MFI2, HAS3, ADAMTS9, HEY1, DIO2 and FGFR3. A functional screen using siRNA suggested roles for MFI2, HEY1 and DIO2 in osteoblastic differentiation of hMSC. Profile B contained genes transiently downregulated by BMP-7, including numerous genes associated with cell cycle regulation. Follow-up studies confirmed that BMP-7 attenuates cell cycle progression and cell proliferation during early osteoblastic differentiation. Profile C, comprised of genes continuously downregulated by BMP-7, exhibited strong enrichment for genes associated with chemokine/cytokine activity. Inhibitory effects of BMP-7 on cytokine secretion were verified by analysis of enriched culture media. Potent downregulation of CHI3L1, a potential biomarker for numerous joint diseases, was also observed in Profile C. A focused evaluation of BMP, GDF and BMP inhibitor expression elucidated feedback loops modulating BMP 7 bioactivity. BMP-7 was found to induce BMP-2 and downregulate GDF5 expression. Transient knockdown of BMP-2 using siRNA demonstrated that osteoinductive properties associated with BMP-7 are independent of endogenous BMP-2 expression. Noggin was identified as the predominant inhibitor induced by BMP-7 treatment. Overall, this study provides new insight into key bioactivities characterizing early BMP-7 mediated osteoblastic differentiation. PMID- 19306957 TI - Transmembrane cholesterol migration in planar lipid membranes measured with Vibrio cholerae cytolysin as molecular tool. AB - The rate of transbilayer movement (flip-flop) of cholesterol was estimated using planar bilayers with defined initial asymmetry, formed by the opposing monolayers technique. Vibrio cholerae cytolysin (VCC) was utilized as a molecular tool for measuring the cholesterol concentration in the cis leaflet of asymmetric bilayers. To quantify cholesterol flip-flop in planar lipid bilayers, a mathematical model was developed. It considers both the lateral diffusion rate of cholesterol within each monolayer and the flip-flop rate. The difference in initial and steady-state cholesterol contents in bilayer leaflets was used as a start point. Assuming the lateral diffusion coefficient to be of 1 x 10(-8) cm(2) s(-1), the characteristic time of cholesterol flip-flop at 25 +/- 2 degrees C was estimated as <10 s. PMID- 19306958 TI - Integrin signaling and cell spreading alterations by rottlerin treatment of chick limb bud mesenchymal cells. AB - Endochondral skeletal development begins with the formation of a cartilaginous template where mesenchymal cells aggregate and increase in density prior to their overt differentiation into chondrocytes. Prechondrogenic condensation, in which mesenchymal cells aggregate, requires cell migration and proliferation. However, the molecular mechanisms promoting this aggregation remain to be elucidated. Here, we report that rottlerin suppresses migration and cell surface expression of integrin beta1 in chondrogenic progenitors. Perturbation of integrin beta1 function using an anti-integrin beta1 blocking antibody suppressed the migration of wing bud mesenchymal cells. Furthermore, phosphorylation levels of Src and focal adhesion kinase (FAK) were decreased by rottlerin treatment. Cell treatment with PP2, an inhibitor of Src family kinase, or electroporation of FAK specific siRNA, suppressed cell migration in a wound-healing assay. Cells treated with rottlerin showed decreased phosphorylation of Akt, independent of PKCdelta inhibition. In addition, an Akt inhibitor suppressed the migration of chick limb bud mesenchymal cells. Taken together, our results point to the novel finding that rottlerin may act as a negative regulator for cell migration, an essential step for prechondrogenic condensation, by regulating integrin beta1 signaling at focal adhesion complexes via modulation of Akt activity. PMID- 19306959 TI - Hormonal contraception, sickle cell trait, and risk for venous thromboembolism among African American women. AB - OBJECTIVE: We evaluated the effect of oral and other hormonal contraceptive (HC) use on venous thromboembolism risk among African American women and investigated whether the association was modified by the sickle cell trait. STUDY DESIGN: We report the findings of a case-control study that included 60 African American women with an idiopathic, first episode of venous thromboembolism and 196 African American controls. RESULTS: The odds of current HC use compared with noncurrent use contrasting cases and controls is 3.8 (95% confidence interval [CI], 1.7-8.1; P < .001). Among subjects with sickle cell trait, the odds ratio is higher (odds ratio [OR], 6.7; 95% CI, 1.0-43) than the odds ratio among subjects without sickle cell trait (OR, 2.6; 95% CI, 1.1-6.2), but the difference is not statistically significant. CONCLUSION: This study provides persuasive evidence that hormonal contraceptive use increases venous thromboembolism risk among African American women and that the increase in risk may be larger among women with sickle cell trait. PMID- 19306960 TI - The role of bromocriptine in peripartum cardiomyopathy. PMID- 19306963 TI - The effect of barusiban, a selective oxytocin antagonist, in threatened preterm labor at late gestational age: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: The objective of the study was to compare barusiban with placebo in threatened preterm labor. STUDY DESIGN: This was a randomized, double-blind, placebo-controlled, multicenter study. One hundred sixty-three women at 34-35 weeks plus 6 days, and with 6 or more contractions of 30 seconds duration during 30 minutes, cervical length 15 mm or less, and cervical dilatation > 1 and < 4 cm were randomized to a single intravenous bolus of barusiban (0.3, 1, 3, or 10 mg) or placebo. The primary endpoint was percentage of women who did not deliver within 48 hours. RESULTS: None of the barusiban doses reduced the number of uterine contractions compared with placebo. There was no significant difference in the percentage of women who did not deliver within 48 hours (72% placebo and 65-88% barusiban groups; P = .21-.84). Barusiban was not associated with an adverse safety profile in the woman, fetus, neonate, or infant. CONCLUSION: An intravenous bolus of barusiban was no more effective than placebo in stopping preterm labor in pregnant women at late gestational age. PMID- 19306964 TI - Metformin usages in women with heart failure. PMID- 19306961 TI - Maternal plasma concentrations of the soluble tumor necrosis factor receptor 2 are increased prior to the diagnosis of preeclampsia. AB - OBJECTIVE: Soluble receptor levels of tumor necrosis factor (sTNF-R)-1 and -2 are increased during preeclampsia. We postulated the increase preceded overt disease. STUDY DESIGN: Archived plasma from the Eunice Kennedy Shriver National Institute of Child Health and Human Development aspirin to prevent preeclampsia in high risk women trial were used to measure serial sTNF-R1 and sTNF-R2 (enrollment, 24 28 week's gestation) in 986 women (577 also sampled at 34-38 weeks). RESULTS: Preeclampsia incidence was 21.2%. sTNF-R2 levels were higher at enrollment (P = .02) and weeks 24-28 (P = .01) in women who eventually developed preeclampsia. The magnitude of increase from baseline of both receptors was significantly greater in women who developed preeclampsia in the future. Women with week 24-28 sTNF-R2 levels in the highest quartile had significantly increased odds to develop preeclampsia (P = .03 vs quartile 1). This association was observed in the placebo but not the aspirin arm (P 65 years), and male gender (69.9% vs 72.1%; all P < .001) all had lower rates. Compared to 2005, more patients were current with guidelines (71% vs 64%) and colonoscopic screening (83% vs 71%). CONCLUSIONS: Although ethnicity-, gender-, and age-related disparities were observed, screening rates are improved in an equal access healthcare system. PMID- 19306972 TI - Deep and organ/space infections in patients undergoing elective colorectal surgery: incidence and impact on hospital length of stay and costs. AB - BACKGROUND: The reported incidence of infection complicating elective colorectal surgery (ECS) is 11% to 26%. We evaluated length of stay (LOS) and expense associated with such infections, which heretofore remain unexplored. METHODS: We reviewed 1127 ECS procedures from October 2005 to may 2007 to identify infected case subjects (n = 46). Data were obtained by way of chart abstraction and administrative database review. A case-control study evaluated LOS and actual accounting costs for case subjects versus uninfected control subjects (n = 46). Logistic regression determined risk factors for infection. RESULTS: Infection incidence was 4.1%. Mean +/- SD LOS and costs were greater for case than control subjects: 21 +/- 15 days compared with 6 +/- 4 days (P < .001) and $42,516 +/- 39,972 compared with $10,999 +/- $7,122 (P < .001). Procedure type, infection, chronic obstructive pulmonary disease, increased age, and nonsmoking status predicted greater LOS and costs. Infection risk factors included duration of procedure > or =3 hours, male sex, higher American Society of Anesthesiologists (ASA) score, low baseline hematocrit, and indication for surgery of regional enteritis/ulcerative colitis. COMMENTS: Infection development after ECS is infrequent in our population, but it results in significantly poorer outcomes. Vigilant adherence to preventive guidelines, including those for antibiotic prophylaxis, is warranted. PMID- 19306973 TI - Alpine ski and snowboarding traumatic injuries: incidence, injury patterns, and risk factors for 10 years. AB - BACKGROUND: Alpine skiing and snowboarding are popular winter sports in Canada. Every year participation in these activities results in traumatic injury. The purpose of this study was to identify the incidence and injury patterns, as well as risk factors associated with ski and snowboarding injuries. METHODS: A comprehensive 10-year retrospective review of Alpine ski and snowboarding injuries from 1996 to 2006 was conducted. The Alberta Trauma Registry was used as the primary source of data. RESULTS: A total of 196 patients (56.6% skiers, 43.4% snowboarders) were identified as having major traumatic injuries (Injury Severity Score, >or=12). Forty-three patients required intensive care unit support. The majority of injuries were related to falls and collisions with natural objects. Head injuries were most common, followed by chest, spinal, and extremity trauma. Seventy-nine patients required emergency surgery. CONCLUSIONS: Skiing and snowboarding represent activities with high potential for traumatic injury. Safety initiatives should be developed to target this population. PMID- 19306974 TI - Surgical outcomes following pancreatic resection at a low-volume community hospital: do all patients need to be sent to a regional cancer center? AB - BACKGROUND: The only curative option for patients with pancreatic cancer is surgical resection. The potential for significant morbidity and mortality following these procedures along with short-term survival benefit has called into question the role of surgery in this disease. Several recent reports have shown that morbidity, mortality, and survival can be improved if these pancreatic resections are performed at centers where large volumes of cases are done annually. METHODS: A retrospective review of the tumor registry from 1994 to 2003 identified 242 cases of pancreatic cancer diagnosed and/or treated at our institution. During this period, 31/242 (13%) patients underwent surgical resection. Patients' charts were reviewed for diagnosis, stage of tumor, presenting symptoms, surgery, length of stay, and survival. Morbidity and mortality rates were calculated for all patients. RESULTS: Thirty-one resections were performed in 16 males and 15 females. The median age at presentation was 69 years. The most common presenting symptom was painless jaundice. A pancreaticoduodenectomy was the most common procedure (n = 24), while 7 distal pancreatectomies were also performed. Eight surgeons performed the 31 resections with one surgeon performing 12 of the cases. The median length of stay was 16 days. Complications arose in 15/31 (48%) patients. There was no 30-day surgical or in-hospital mortality. CONCLUSIONS: Major pancreatic surgery can be performed safely at community hospitals. It is imperative that each hospital is responsible for providing morbidity and mortality figures related to pancreatic procedures performed at their institution. In this changing climate of reimbursement and pay for performance, institutions that do not do this may be required to send these cases to regional centers. PMID- 19306975 TI - Gastric malignancies in breast cancer survivors: pathology and outcomes. AB - BACKGROUND: As the number of breast cancer survivors increases, the appearance of second malignancies and unusual metastatic patterns likely also is increasing. In particular, we and others have observed gastric malignancies in breast cancer survivors. METHODS: We reviewed 3 regional hospital system tumor databases, comprising 19,049 analytic breast cancer cases, to determine the number, types, and outcomes of subsequent gastric malignancies. RESULTS: Twenty-eight patients developed subsequent gastric malignancies, representing .15% of breast cancer survivors; 82% of patients had gastric symptoms. Overall survival for the cohort was 39%. Twenty-four patients (86%) had gastric primaries and 13 died of their second cancers. Four patients had gastric metastases; all had lobular histology in both their primary tumors and metastatic lesions. Five patients had gastrointestinal stromal tumors; all patients underwent resection and currently are alive. CONCLUSION: Gastric symptoms in breast cancer survivors may represent malignant lesions, often second primaries. All gastric metastases in our series were of lobular histology. PMID- 19306976 TI - Secure placement of a peripancreatic drain after a distal pancreatectomy. AB - BACKGROUND: A peripancreatic drain that is placed after a distal pancreatectomy sometimes migrates and becomes ineffective postoperatively. We devised a new drainage method with fixation of the tip of a peripancreatic drain using a loose loop of an absorbable suture. METHODS: This retrospective study was performed on 84 consecutive patients who underwent a distal pancreatectomy followed by peripancreatic drainage with (n = 31) or without (n = 53) fixation. RESULTS: The fixed drain remained in place postoperatively and was removed easily when the drainage became unnecessary. Pancreatic fistula developed in 4 patients with and 11 patients without drain fixation, the incidence between the patients. None with and 7 patients without fixation required additional drainage (interventional or surgical) for pancreatic fistula, the difference being significant. Time to resolution of pancreatic fistula tended to be shorter after drain fixation than after nonfixation. CONCLUSIONS: Fixation of the tip of a peripancreatic drain is a simple but useful technique for effective drainage after distal pancreatectomy. PMID- 19306977 TI - Recent trends and predictors in immediate breast reconstruction after mastectomy in the United States. AB - INTRODUCTION: Immediate breast reconstruction after mastectomy has increased in frequency during the past decade, but the socioeconomic and patient factors have yet to be fully identified. METHODS: Data were analyzed from the Nationwide Inpatient Sample from 1999 to 2003 using International Classification of Disease 9 codes to identify patients undergoing immediate breast reconstruction. Regression analyses were used to examine predictive variables for immediate breast reconstruction after mastectomy. RESULTS: Between 1999 and 2003, 469,832 patients underwent mastectomy. Immediate breast reconstruction occurred in 110,878 patients, yielding a 5-year average rate of 23.6% (range of 22.2% to 25.3%). Independent predictors of immediate breast reconstruction after mastectomy include private insurance, hospital in an urban location, teaching hospital, white race, hospital region in the south, age between the 3rd and 6th decades, and low number of comorbidities. CONCLUSIONS: Immediate breast reconstruction after mastectomy is still not commonly performed in the United States. Socioeconomic and geographic factors play a significant role in whether patients undergo immediate reconstruction. PMID- 19306978 TI - The OPTICC trial: a multi-institutional study of occult pneumothoraces in critical care. AB - BACKGROUND: The management of pneumothoraces detected on CT but not on supine chest radiographs remains controversial, especially in those undergoing positive pressure ventilation (PPV) who are at risk for complications with both observation and treatment. Previous limited study yielded confusion regarding the need for routine drainage of these occult pneumothoraces (OPTXs). We conducted a pilot study at 2 trauma centers to address the feasibility and safety of randomizing traumatized patients undergoing PPV to drainage or observation. METHODS: Stable mechanically ventilated (or en route to surgery) adults with OPTXs were identified at 2 centers (Calgary and Quebec). Patients were randomized to observation (unless drainage became clinically indicated) or to chest drainage. Episodes of respiratory distress (need for thoracostomy tube, acute/sustained increase in oxygen requirements, difficulty in achieving adequate ventilation and self-reported distress) and subsequent imaging abnormalities were recorded until discharge. RESULTS: From August 2006 to April 2008, 24 trauma patients were enrolled (17 Calgary and 7 Quebec), with 2 later exclusions (final CT found no OPTX). Thirteen patients (59%) were randomized to observation, 9 to drainage (41%). Four observed (31%) later had chest tubes placed nonurgently for worsening OPTXs/effusions; none with increased morbidity. Overall rates of respiratory distress (drainage: 33%, observation: 41%) and mortality (drainage: 22%, observation: 15%) were similar across groups, as were median intensive care unit (drainage: 3, observation: 4) and in-hospital days (drainage: 10, observation: 16). CONCLUSIONS: With no important differences in morbidity, the OPTICC pilot lays the foundation for a future definitive trial comparing drainage or observation in posttraumatic OPTXs requiring PPV. PMID- 19306979 TI - Differential molecular changes in patients with asymptomatic long-segment Barrett's esophagus treated by antireflux surgery or medical therapy. AB - BACKGROUND: The Barrett's to adenocarcinoma sequence is characterized by molecular changes including activation of nuclear factor-kappaB (NF-kappaB) and related cytokines. In this observational nonrandomized study this molecular environment was compared in matched asymptomatic cohorts who had undergone either fundoplication or therapy with proton pump inhibitors (PPIs). METHODS: Asymptomatic patients with long-segment Barrett's esophagus had endoscopic biopsy specimens taken from 2 cm below the squamocolumnar junction for measurement of activated NF-kappaB and a panel of cytokines and growth factors. RESULTS: Thirty seven patients were recruited (surgical: n = 18, medical: n = 19). The mean patient age was 51 years, and the mean follow-up period was 5.6 years. There were no differences in the length of Barrett's segment and endoscopic and histopathologic features in both groups. Mean activated NF-kappaB p50 and p65 subunits, interleukin (IL)-1alpha, IL-1beta, and interleukin-8 levels, were significantly (P < .05) lower in the surgically treated group. CONCLUSIONS: This study provides proxy support to the thesis that antireflux surgery may provide an environment that is less inflammatory and tumorigenic than that observed in medically treated patients. PMID- 19306980 TI - Diagnostic value of hyperbilirubinemia as a predictive factor for appendiceal perforation in acute appendicitis. AB - BACKGROUND: Appendiceal perforation in patients with acute appendicitis may cause a variety of potentially life-threatening complications. Escherichia coli endotoxin has been shown to impact physiological bile flow in vivo. This had led to the theory that hyperbilirubinemia in patients with appendicitis may have a predictive potential for the preoperative diagnosis of appendiceal perforation. The aim of this retrospective study was to investigate the diagnostic value of hyperbilirubinemia as a preoperative laboratory marker for appendiceal perforation in patients with acute appendicitis. METHODS: We identified 538 patients (306 female; 232 male, mean age, 35.6 y) with histologically proved acute appendicitis who underwent laparoscopic or conventional appendectomy between January 2004 and December 2007 in a surgical department of an academic teaching hospital. A retrospective multiple chart review of the medical records including laboratory values and histologic results was conducted. RESULTS: The mean bilirubin level of all patients was .9 mg/dL (+/-.6 SD mg/dL; range, .1-4.3 mg/dL; median, .7 mg/dL). Patients with appendiceal perforation, however, had a mean bilirubin level of 1.5 mg/dL (+/-.9 SD mg/dL; range, .4-4.3 mg/dL; median, 1.4 mg/dL), which was significantly higher than those with a nonperforated appendicitis (P < .05). The specificity of hyperbilirubinemia for appendiceal perforation was .86 compared with .55 for white blood count and .35 for C reactive protein. Sensitivity was .7 compared with .81 for white blood count and .96 for C-reactive protein. CONCLUSIONS: Patients with hyperbilirubinemia and clinical symptoms of appendicitis should be identified as having a higher probability of appendiceal perforation than those with normal bilirubin levels. PMID- 19306981 TI - Do patients undergoing parathyroidectomy for primary hyperparathyroidism in San Francisco, CA, and Bursa, Turkey, differ? AB - BACKGROUND: To compare the preoperative clinical symptoms and associated conditions and findings in operations in age- and sex-matched patients with primary hyperparathyroidism (PHPT) living in the two different regions of the world to determine whether PHPT differs in various countries. METHODS: Fifty-two patients from Bursa, Turkey were matched with 52 age and sex matched patients from San Francisco, USA. Patients' preoperative symptoms, biochemical and radiologic findings and surgical procedures were documented. RESULTS: More patients in the American Group (15%) had preoperatively persistant or recurrent hyperparathyroidism, P = 0.016. Serum parathyroid hormone levels were higher in Turkish group (546 +/- 75.33 pg/mL) than in American group (146 +/- 75.33 pg/mL). More Turkish patients had osteoporosis (P < 0.05). The size of parathyroid adenomas was significantly greater in Turkish patients (25.2 +/- 1.18 mm) than in American patients (17.5 +/- 1.18 mm), P < 0.001. CONCLUSIONS: Patients with PHPT from Bursa, Turkey have higher plasma parathyroid hormone levels, larger parathyroid adenomas and more severe bone disease than in age and sex matched patients with PHPT in San Francisco. PMID- 19306982 TI - Controlled tube duodenostomy in the management of giant duodenal ulcer perforation: a new technique for a surgically challenging condition. AB - BACKGROUND: Giant duodenal ulcer (GDU) perforation remains an extremely uncommon but a rather challenging condition to manage wherein routine surgical procedures are fraught with an extremely high incidence of failure and mortality. It therefore follows that this condition must be identified and managed differently at laparotomy than are most duodenal perforations. We describe a method by which to deal with this condition using triple-tube-ostomy. METHODS: In a prospective setting, 20 patients underwent surgery using the technique described in the article. During the same period, 20 patients with GDU perforation, who were managed in the conventional manner, were evaluated. The outcomes of the 2 groups were compared. RESULTS: The success rate was 100% in the study group compared with 30% in the control patients. CONCLUSIONS: Based on the ease of the technique and the high success of the procedure in our experience in this select group, we recommend this procedure for the management of GDU perforation as a safe, reliable, and easy technique to learn. PMID- 19306983 TI - Development of a comprehensive vascular skills assessment for surgical trainees. AB - BACKGROUND: Although vascular skills are important to general surgeons, vascular surgery has become a separate specialty, and therefore, there may be an erosion of vascular skills acquired by general surgical trainees. The purpose of this study was to develop a reliable and valid comprehensive vascular skills assessment (CVSA) of both knowledge and technical skills. METHODS: Twenty-four of 38 general surgical residents at the University of British Columbia completed a two-part CVSA consisting of a written examination and a series of 4 technical stations in a skills laboratory. Technical performance was rated using validated scales. RESULTS: The mean overall CVSA score was 50%. The CVSA demonstrated construct validity, with improvement in scores with increasing postgraduate year level (P = .01). The overall reliability (Cronbach's alpha) was .90. CONCLUSIONS: The CVSA developed in this study is a comprehensive assessment of vascular skills that is both valid and reliable. It offers an objective and feasible assessment of general surgical trainees' vascular skills. PMID- 19306984 TI - Prophylactic mesh to prevent incisional hernia: a note of caution. AB - BACKGROUND: Ventral hernia is a common complication of open Roux-en-Y gastric bypass (RYGB). The aim of this study was to determine whether prophylactic mesh placement during RYGB would reduce the incidence of postoperative hernias. METHODS: Obese patients undergoing RYGB by a single surgeon had prosthetic mesh placed in a subfascial location at the conclusion of the procedure. The incidences of recurrent hernia and morbidity associated with the placement of mesh were assessed. RESULTS: Sixteen patients underwent RYGB with prophylactic mesh placement over 6 months. The average preoperative body mass index was 46.6 kg/m(2). Half of the patients were diabetics. None were smokers. During mean follow-up of 6 months, 4 patients (25%) required mesh excision, 3 for infection and 1 for a persistently symptomatic seroma. One patient was explanted incidentally in the course of reexploration for intractable nausea and vomiting. Another developed an incisional hernia despite prophylactic mesh. CONCLUSIONS: In the investigators' experience, the use of prophylactic new-generation mesh at the time of open RYGB led to an unacceptable rate of local complications. They caution against this technique in patients undergoing open RYGB. PMID- 19306985 TI - Re: The survival impact of the choice of surgical procedure after ipsilateral breast cancer recurrence. PMID- 19306986 TI - Effective use of laparoscopy for long-term peritoneal dialysis access. AB - BACKGROUND: Laparoscopy is an underused modality for peritoneal dialysis access procedures. The strengths of laparoscopy are that it can both prevent and resolve the common mechanical problems that adversely effect dialysis catheter outcomes. METHOD: Laparoscopically enabled catheter implantation and rescue procedures included rectus sheath catheter tunneling, omentopexy, adhesiolysis, resection of epiploic appendices, colopexy, salpingectomy, and appendectomy. Using these techniques, the outcomes of 428 laparoscopically implanted catheters were studied. RESULTS: During a mean follow-up of 21.6 months, mechanical obstruction complicated 3.7% of implantation procedures. The incidence of pericatheter leak was 2.6%. There were no occurrences of pericatheter hernia or subcutaneous cuff extrusion. Laparoscopic salvage procedures limited losses from mechanical catheter problems to .9%. Cumulative revision-free and assisted catheter survival probabilities for loss from mechanical complications at 5 years were .96 and .99, respectively. COMMENTS: Because it is enabled by techniques not available to other catheter-placement methods, laparoscopy produces superior outcomes. PMID- 19306987 TI - Quality of surgery for rectal carcinoma: comparison between open and laparoscopic approaches. AB - BACKGROUND: Macroscopic evaluation of a tumor specimen is an independent prognostic factor of oncologic outcome after total mesorectal excision (TME) for rectal cancer. This study aimed to assess macroscopic quality of specimens acquired after laparoscopic versus open TME in patients with low rectal cancer. PATIENTS AND METHODS: Seventy-two patients with low rectal cancer underwent TME either by open (n = 39) or laparoscopic (n = 33) approach. In all specimens, the cut edge of the peritoneal reflection at the anterior mid-rectum, the Denonvillier's fascia, the visceral fascia covering the mesorectum both posteriorly and laterally, and the bowel wall below the mesorectum were macroscopically assessed. RESULTS: Colorectal anastomoses were located significantly lower in the laparoscopic than in the open group (P < .001). The Denonvillier's fascia was violated in 7 patients after open surgery (P = .01). A significantly more complete TME with intact visceral pelvic fascia was performed after laparoscopy compared with open surgery (P = .025). CONCLUSIONS: Laparoscopy offers a macroscopically more complete specimen after TME for rectal cancer than the open approach because it offers a better view in the pelvis. PMID- 19306988 TI - Timing may be important, but methodology is everything: a commentary on parker et Al. PMID- 19306990 TI - Allelic variation in the serotonin transporter promoter modulates cortical excitability. AB - BACKGROUND: Transcranial magnetic stimulation (TMS) is a noninvasive procedure that may be used to study individual differences in motor cortex excitability. Such differences are assumed to reflect serotonergic and other inputs to the motor cortex. METHODS: Here we investigate the impact of a functional polymorphism in the promoter region of the 5-hydroxytryptamine transporter gene (5-HTTLPR) on measures of motor cortex excitability. RESULTS: Sixty healthy subjects carrying one or two copies of the short 5-HTTLPR allele (s/s and s/l) showed a significant reduction in short intracortical inhibition (SICI, p = .012) and an increased cortical silent period (p = .042) compared with 60 age- and sex matched individuals homozygous for the long allele (l/l). In contrast, motor threshold and intracortical facilitation did not differ significantly between groups. CONCLUSIONS: These results provide further evidence of a role for serotonergic transmission in the modulation of cortical excitability. Differential effects on the measures under study suggest a pattern of prioritization in bioamine regulation of cortical inhibition. PMID- 19306991 TI - Conformational studies of the capsular polysaccharide produced by Neisseria meningitidis group A. AB - The effect of different cations on the conformational and morphological properties of the capsular polysaccharide produced by Neisseria meningitidis group A was investigated. Circular dichroism studies showed that the presence of Na(+), NH4+ or Ca(2+) ions induced different local conformations of the polysaccharide chain through interactions with the phosphodiester group bridging the saccharide residues in the polymer chain. Atomic force microscopy experiments confirmed that the morphology of the polysaccharide chains was different depending on the nature of the counterion. Ammonium ions were associated with the presence of single polymer chains in an elongated conformation, whereas sodium ions favored the folding of the chains into a globular conformation. The addition of calcium ions produced the aggregation of a limited number of globular polysaccharide chains to form a 'toroidal-like' structure. PMID- 19306992 TI - Dysfunctional memory CD8+ T cells after priming in the absence of the cell cycle regulator E2F4. AB - The transcriptional repressor E2F4 is important for cell cycle exit and terminal differentiation in epithelial cells, neuronal cells and adipocytes but its role in T lymphocytes proliferation and memory formation is not known. Herein, we investigated the function of E2F4 protein for the formation of functional murine memory T cells. Murine transgenic CD8+ T cells were infected in vitro with lentivirus vector expressing a shRNA targeted against E2F4 followed by in vitro stimulation with SIINFEKL antigenic peptide. For in vivo assays, transduced cells were injected into congenic mice which were then infected with HSV-OVA. The primary response, memory formation and secondary stimulation were determined for CD8+ lentivirus transduced cells. In the absence of E2F4 cell cycle repressor, activated CD8+ T cells underwent intensive proliferation in vitro and in vivo. These cells had the ability to differentiate into memory cells in vivo, but they were defective in recall proliferation. We show that transient suppression of E2F4 during CD8+ T cell priming enhances primary proliferation and has a negative effect on secondary stimulation. These findings demonstrate that the cell cycle repressor E2F4 is essential for the formation of functional memory T cells. A decrease in CD8+ T-lymphocyte compartment would diminish our capacity to control viral infections. PMID- 19306993 TI - The distinct response of gammadelta T cells to the Nod2 agonist muramyl dipeptide. AB - Purified gammadelta T cells are primed directly in response to pathogen associated molecular patterns (PAMPs) to better respond to secondary signals and increase expression of chemokine and activation-related genes. Transcripts encoding the innate receptor Nod2 were detected in bovine and human gammadelta T cells. Nod2 is the intracellular receptor for muramyl dipeptide (MDP), functions in regulating innate activities, and was thought to be expressed primarily in APCs. The response of gammadelta T cells to MDP was analyzed by microarray, Q PCR, proteome array and functional priming assays. MDP had a consistent priming effect on gammadelta T cells, characterized by changes in transcripts and enhanced proliferation response to secondary signaling. Knockdown experiments implicated Nod2 as the receptor for MDP in gammadelta T cell-enriched bovine PBLs. The results indicate priming of gammadelta T cells by MDP, and offer definitive evidence of the expression of functional Nod2 in gammadelta T cells. PMID- 19306994 TI - Autologous dendritic cells loaded with apoptotic tumor cells induce T cell mediated immune responses against breast cancer in vitro. AB - Dendritic cell (DCs) based immunotherapy has received increased interest in the treatment of specific malignancies including breast cancer. In this in vitro study, T cell responses, which are induced by monocyte-derived DCs pulsed with apoptotic breast tumor cells (ApTC), were analyzed in terms of proliferation, specific cytotoxicity, and cytokine release. Nylon wool-enriched T lymphocytes from five patients with breast cancer stimulated with monocyte-derived DCs pulsed with apoptotic tumor cells in vitro and their proliferation response were analyzed by [(3)H] thymidine uptake and specific cytotoxic activity of tumor antigen-primed T cells after three rounds of weekly stimulation by flow cytometry. Interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) cytokine release assay was carried out 24h after the last stimulation. The supernatant from primed T cells was collected and analyzed using commercially available ELISA kits. T cell proliferation assays revealed that DCs pulsed with apoptotic tumor cell could stimulate an autologous T cell proliferation response with stimulation indices of 5-21. The T cell-mediated cytotoxicity assay demonstrated that tumor antigen-primed T cells could kill significantly more autologous tumor cells than normal cells (P<0.05). These cells had variable amounts of cytotoxic activity against K562 cells. Primed T cells released both IFN-gamma and IL-4 in response to re-stimulation by antigen-pulsed DCs, but were dominated by IFN-gamma production in two out of five patients and IL-4 production in three out of five patients. In conclusion, our results suggested that DCs pulsed with apoptotic breast tumor cells could elicit effective specific antitumor T cell responses in vitro. Therefore, vaccination with DCs pulsed with apoptotic tumor cells may be considered as a novel strategy for immunotherapy of patients with breast cancer refractory to standard modalities. PMID- 19306995 TI - The cognitive architecture for chaining of two mental operations. AB - A simple view, which dates back to Turing, proposes that complex cognitive operations are composed of serially arranged elementary operations, each passing intermediate results to the next. However, whether and how such serial processing is achieved with a brain composed of massively parallel processors, remains an open question. Here, we study the cognitive architecture for chained operations with an elementary arithmetic algorithm: we required participants to add (or subtract) two to a digit, and then compare the result with five. In four experiments, we probed the internal implementation of this task with chronometric analysis, the cued-response method, the priming method, and a subliminal forced choice procedure. We found evidence for an approximately sequential processing, with an important qualification: the second operation in the algorithm appears to start before completion of the first operation. Furthermore, initially the second operation takes as input the stimulus number rather than the output of the first operation. Thus, operations that should be processed serially are in fact executed partially in parallel. Furthermore, although each elementary operation can proceed subliminally, their chaining does not occur in the absence of conscious perception. Overall, the results suggest that chaining is slow, effortful, imperfect (resulting partly in parallel rather than serial execution) and dependent on conscious control. PMID- 19306996 TI - Inferring agency from sound. AB - In three experiments we investigated how people determine whether or not they are in control of sounds they hear. The sounds were either triggered by participants' taps or controlled by a computer. The task was to distinguish between self control and external control during active tapping, and during passive listening to a playback of the sounds recorded during the active condition. Experiment 1 required detection of a change in control mode within trials. Experiments 2 and 3 introduced a simple rhythm reproduction task that requires discrimination of control modes between trials. The results demonstrate that both sensorimotor cues and perceptual cues are used to infer agency. In addition, there may be further influences of cognitive expectation and/or multimodal integration. In accordance with hierarchical models of intention [e.g., Pacherie, E. (2008). The phenomenology of action: A conceptual framework. Cognition, 107, 179-217] this suggests that the sense of agency is not situated on one specific level of action control but subject to multiple influences. PMID- 19306997 TI - Determination of patulin in fruit juice and dried fruit samples by in-tube solid phase microextraction coupled with liquid chromatography-mass spectrometry. AB - A simple and sensitive method for the determination of patulin in fruit juice and dried fruit samples was developed using a fully automated method consisting of in tube solid-phase microextraction (SPME) coupled with liquid chromatography-mass spectrometry (LC-MS). Patulin was separated within 5 min by high-performance liquid chromatography using a Synergi MAX-RP 80A column and water/acetonitrile (80/20, v/v) as the mobile phase. Electrospray ionization conditions in the negative ion mode were optimized for MS detection of patulin. The pseudo molecular ion [M-H](-) was used to detect patulin in selected ion monitoring (SIM) mode. The optimum in-tube SPME conditions were 25 draw/eject cycles of 40 microL of sample using a Carboxen 1006 PLOT capillary column as an extraction device. The extracted patulin was readily desorbed from the capillary by passage of the mobile phase, and no carry-over was observed. Using the in-tube SPME LC-MS with SIM method, good linearity of the calibration curve (r=0.9996) was obtained in the concentration range of 0.5-20 ng/mL using (13)C(3)-patulin as an internal standard, and the detection limit (S/N=3) of patulin was 23.5 pg/mL. The in-tube SPME method showed >83-fold higher sensitivity than the direct injection method (10 microL injection volume). The within-day and between-day precision (relative standard deviations) were below 0.8% and 5.0% (n=6), respectively. This method was applied successfully for the analysis of fruit juice and dried fruit samples without interference peaks. The recoveries of patulin spiked into apple juice were >92%, and the relative standard deviations were <4.5%. Patulin was detected at ng/mL levels in various commercial apple juice samples. PMID- 19306998 TI - A new version of an additive scheme for the prediction of gas chromatographic retention indices of the 211 structural isomers of 4-nonylphenol. AB - Control of environmental pollution by 4-nonylphenols (4-NP) and effective risk assessment concerning these xenoestrogens requires the identification of the individual isomers contained in the technical mixtures of 4-NP. A new approach is presented here which supports the identification of these compounds by a combination of experimentally determined gas chromatographic retention indices (I) of reference 4-NP isomers and calculated I-values. In addition to experimental indices, the I-prediction algorithm includes a new version of an additive scheme. The I-values of all structural 4-NP isomers are calculated on the basis of experimentally determined indices of a few available 4-NP isomers and the known retention indices of 75 iso-decanes. A mean deviation of +/-11 index units between predicted and experimental I-values demonstrates the feasibility of the new approach. The predicted I-values provide information on the structure of 4-nonylphenol isomers in the technical mixture which has not been considered before. Furthermore, a novel line-coding system is proposed to describe the structure of isomeric 4-NPs and to initiate a current database for the endocrine-disrupting 4-nonylphenols. PMID- 19306999 TI - Novel multiwalled carbon nanotubes-polyaniline composite film coated platinum wire for headspace solid-phase microextraction and gas chromatographic determination of phenolic compounds. AB - A novel multiwalled carbon nanotubes-polyaniline composite (MWCNTs-PANI) film coated platinum wire was fabricated through electrochemical deposition. The coating was characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectrophotometry and thermogravimetry. It was found that the coating was porous and had large specific area and adsorption capacity; in the composite MWCNTs and polyaniline interacted with each other and the film kept stable up to 320 degrees C. The as-made fiber was used for the headspace solid-phase microextraction (HS-SPME) of some phenolic compounds (i.e. 2 chlorophenol, 2,4-dichlorophenol, 2-methylphenol, 3-methylphenol, 2,6 dimethylphenol, 2-nitrophenol), followed by gas chromatographic analysis. The MWCNTs-PANI coating showed better analytical performance than PANI. Under the optimized conditions, the detection limits were 1.89-65.9 ng L(-1), the relative standard deviations (RSDs) were 2.7-6.5% for six successive measurements with single fiber, the RSDs for fiber-to-fiber were 5.2-12.4%, the linear ranges exceeded two magnitudes with correlation coefficient above 0.992. The fiber could be used for more than 250 times without decrease of efficiency. The proposed method was successfully applied to the extraction and determination of phenolic compounds in water sample, and the recoveries were 87.7-111.5% for different analytes. In addition, the fiber also presented advantages of easy preparation and low cost. Therefore, it is a promising SPME fiber. PMID- 19307000 TI - Rapid and efficient purification of positron emission tomography probes by hydrophilic interaction chromatography. AB - A rapid and efficient preparative high-performance liquid chromatographic procedure was established to purify short-lived positron emission tomography radio-probes. This method is based on hydrophilic interaction chromatography utilizing a semi-preparative silica column (10 mm I.D.) and a high volatile organic mobile phase (>90% acetonitrile). In nine different radiopharmaceuticals studied, six compounds could be separated from the unlabeled precursor with good resolution and faster elution than its precursor. These characteristics enabled significant shortening of the separation and evaporation processes in the manufacture of short-lived radiopharmaceuticals. Several (11)C radiopharmaceuticals could be prepared within one half-life of carbon-11 (20.4 min), including radiosynthesis, purification and formulation steps with sufficient radiochemical/chemical purity and high levels of radioactivity/specific radioactivity. PMID- 19307001 TI - Analysis of pyridylaminated oligosaccharides using liquid chromatography-mass spectrometry with a monolithic capillary column. AB - We examined the utility of a monolithic capillary column in the analysis of pyridylaminated oligosaccharides. Fluorescence detection and mass spectrometry were used to monitor a series of oligosaccharides. Although the total-ion chromatogram appeared similar to that obtained with fluorescence detection, the sensitivity of this technique was limited, especially in the case of smaller oligosaccharides. This limitation was overcome by applying selected ion current monitoring. Further, the capillary column also exhibited good reproducibility. We showed that the retention times obtained by using the monolithic capillary column could be converted into the standard data to enable comparison of the experimental data with the existing data. Furthermore, our studies revealed an important difference in the separation profile, i.e., the monolithic capillary column could resolve smaller oligosaccharides to a greater extent. PMID- 19307002 TI - [Publication of randomized clinical trials: Current existing legislations to guarantee their validity, reliability and transparency]. PMID- 19307003 TI - [Atlantoaxial rotatory subluxation caused by wrestling practice]. PMID- 19307004 TI - [Quality of life and survival after surgical treatment of metastatic femoral fractures]. AB - BACKGROUND AND OBJECTIVES: To analyze the results of the surgical treatment of proximal femoral fractures secondary to metastatic bone disease, in terms of quality of life improvement and survival. MATERIAL AND METHOD: A transversal prospective study was carried out during a period of 15 months in which 20 fractures of femur from 19 patients were included, corresponding to 10 imminent fractures (IF) and 10 established fractures. Assessed final outcomes were associated complications, walking type at discharge and change in Karnofsky's scale after surgery. Mortality and survival after operation were also registered. RESULTS: Surgical procedures performed were osteosynthesis (72%) and femoral arthroplasty (28%). With regard to complications, 1 patient died during the intra operatory period and there was 1 failure of ostesyntesis that required re operation. There was an improvement in the quality of life measured according to the Karnofsky scale after the surgery (P=.017). Survival after surgery was 2 months in the group of patients with impending fracture and 5 months in the group of patients with established fracture (P=.816). CONCLUSIONS: Patients who underwent surgery for a femoral fracture secondary to a metastatic disease showed an improvement in the quality of life, according to the Karnofsky scale. Although they represent a group of patients with a short survival, the control of pain and functional improvement justifiy the procedure. PMID- 19307005 TI - Biosynthesis of antimalarial lignans from Holostylis reniformis. AB - Holostylis reniformis biosynthesizes 8-8' linked lignans without 9,9' oxygenation. To elucidate the biosynthetic pathways to these lignans, the reputed precursors [U-(14)C]phenylalanine, [9-(3)H(1)]coniferyl alcohol, and [9 (3)H(1)]isoeugenol were administered to roots of the plant, which led to the incorporation of (3)H and (14)C into ten 2,7' linked-lignans (aryltetralone lignans) and two 7,7'-epoxylignans (furan lignans). These administration experiments demonstrated that the lignans were propenylphenol-derived and that H. reniformis can exhibit regioselective control over radical-radical coupling (via isoeugenol radicals). Regiospecific control over propenylphenol-derived lignan biosynthesis was observed, together with diastereoselective control of C2-C7' bond formation for the aryltetralone lignans (7'R). These experiments provide evidence that isoeugenol is a biosynthetic intermediate to the aryltetralone and furan lignans. PMID- 19307006 TI - [A mandibular tumor in a six-year-old girl...]. PMID- 19307007 TI - Comment on: Frequency of enteric protozoan parasites among patients with gastrointestinal complaints in medical centers of Zahedan, Iran. PMID- 19307008 TI - A conserved carboxy-terminal domain in the major tegument structural protein VP22 facilitates virion packaging of a chimeric protein during productive herpes simplex virus 1 infection. AB - Recombinant virus HSV-1(RF177) was previously generated to examine tegument protein VP22 function by inserting the GFP gene into the gene encoding VP22. During a detailed analysis of this virus, we discovered that RF177 produces a novel fusion protein between the last 15 amino acids of VP22 and GFP, termed GCT VP22. Thus, the VP22 carboxy-terminal specific antibody 22-3 and two anti-GFP antibodies reacted with an approximately 28 kDa protein from RF177-infected Vero cells. GCT-VP22 was detected at 1 and 3 hpi. Examination of purified virions indicated that GCT-VP22 was incorporated into RF177 virus particles. These observations imply that at least a portion of the information required for virion targeting is located in this domain of VP22. Indirect immunofluorescence analyses showed that GCT-VP22 also localized to areas of marginalized chromatin during RF177 infection. These results indicate that the last fifteen amino acids of VP22 participate in virion targeting during HSV-1 infection. PMID- 19307009 TI - The high-risk HPV E6 oncoprotein preferentially targets phosphorylated nuclear forms of hDlg. AB - High-risk mucosal HPV E6 oncoproteins target a number of PDZ domain-containing substrates for proteasome mediated degradation. One of these, Discs Large (Dlg), is involved in the regulation of cell polarity and proliferation control. Previous studies had suggested that Dlg when hyperphosphorylated by osmotic shock, or when present in the nucleus could be preferentially targeted by E6. In this study we use phospho-specific antibodies directed against Dlg phosphorylated at residues S158 and S442 to show that these two observations are, in fact, linked. Dlg, when phosphorylated on S158 and S442 by CDK1 or CDK2, shows a preferential nuclear accumulation. However, these forms of Dlg are absent in cells derived from HPV-induced cervical cancers. Upon either proteasome inhibition or siRNA ablation of E6 expression, we see specific rescue of these phosphorylated forms of Dlg. These results demonstrate that nuclear forms of Dlg phosphorylated on its CDK phospho-acceptor sites has enhanced susceptibility to E6-induced degradation and place previous studies on the stress-induced phosphorylation of Dlg into a relevant biological context. PMID- 19307010 TI - Receptor usage of a newly emergent adenovirus type 14. AB - Recently, cases of severe respiratory illness in military and civilian populations have been associated with a new genomic variant of adenovirus (Ad) serotype 14, designated Ad14a. Compared to the Ad14 reference strain (de Wit), this new virus had a deletion of two amino acid residues in the fiber protein knob. Here we tested whether this mutation changed receptor usage of Ad14a compared to Ad14-de Wit. Competition studies with radio-labeled viruses revealed that both Ad14-de Wit and Ad14a used the same receptor which is hitherto unknown. We also found that recombinant fiber knobs only partially blocked attachment of Ad14a, indicating that virus capsid proteins other than the fiber are involved in infection. PMID- 19307011 TI - Combination of in vitro bioassays for the determination of cytotoxic and genotoxic potential of wastewater, surface water and drinking water samples. AB - In this study we evaluated genotoxicity and cytotoxicity of native samples of wastewaters (15 samples), surface waters (28 samples) and potable waters (8 samples) with the SOS/umuC assay with Salmonella typhimurium TA1535/pSK1002 and MTT assay with human hepatoma HepG2 cells. The genotoxicity of selected samples was confirmed with the comet assay with HepG2 cells. In the SOS/umuC assay 13 out of the 51 samples were genotoxic: two effluent samples from chemical industry; one sample of wastewater treatment plant effluent; two hospital wastewater samples; three river water samples and four lake water samples. Six samples were cytotoxic for HepG2 cells: both effluent samples of chemical industry, two wastewater treatment plant effluent samples, and two river water samples, however, only the chemical industry effluent samples were genotoxic and cytotoxic, indicating that different contaminants are responsible for genotoxic and toxic effects. Comparing genotoxicity of river and lake water samples with the chemical analytical data of the presence of the residues of pharmaceutical and personal care products (non-steroidal anti-inflammatory drugs, UV filters and disinfectants) in these samples, indicated that the presence of UV filters might be linked to the genotoxicity of these samples. The results showed that the application of the bacterial SOS/umuC assay and mammalian cell assays (MTT and comet assay) with HepG2 cells was suitably sensitive combination of assays to monitor genotoxicity and cytotoxicity of native samples of wastewaters and surface waters. With this study we also confirmed that the toxicity/genotoxicity bioassays should be an integral tool in the evaluation of toxicity of complex wastewaters before the release into environment, as well as for the monitoring of surface water quality, providing data useful in risk assessment. PMID- 19307012 TI - PCDD/PCDF pollution in soils and sediments from the Pearl River Delta of China. AB - This study reported detection of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) for 62 soil samples and 3 sediment samples collected from the Pearl River Delta, a rapidly industrialized and highly populated area of South China. The results show that the PCDD/F concentrations ranged from 97.6 to 9,600 ng kg(-1) and that the average total concentrations of PCDD/Fs are 1311, 2504, 1320 and 2335 ng kg(-1)(dw), and 1.24, 3.99, 4.80 and 2.63 ng WHO(1998)-TEQ kg(-1) for the samples collected from remote mountains, suburban areas, industrial areas, and residential/commercial areas, respectively. The WHO(1998) TEQ values calculated for 17 soil samples are higher than 4 ng kg(-1) but lower than 20 ng kg(-1), whereas the WHO(1998)-TEQ values calculated for the remaining 45 samples are lower than 4 ng kg(-1). According to the Canada soil quality guideline, the PCDD/F concentrations in 27.4% of the tested soil samples in this study are greater than the Canadian background concentrations, and the PCDD/F concentrations for the remaining soil samples (73.6%) are lower than the Canadian background concentration. PMID- 19307013 TI - Quantification of bioregeneration of activated carbon and activated rice husk loaded with phenolic compounds. AB - The bioregeneration efficiencies of powdered activated carbon (PAC) and pyrolyzed rice husk loaded with phenol and p-nitrophenol were quantified by oxygen uptake measurements using the respirometry technique in two approaches: (i) simultaneous adsorption and biodegradation and (ii) sequential adsorption and biodegradation. It was found that the applicability of the simultaneous adsorption and biodegradation approach was constrained by the requirement of adsorption preceding biodegradation in order to determine the initial adsorbent loading accurately. The sequential adsorption and biodegradation approach provides a good estimate of the upper limit of the bioregeneration efficiency for the loaded adsorbent in the simultaneous adsorption and biodegradation processes. The results showed that the mean bioregeneration efficiencies for PAC loaded with phenol and p-nitrophenol, respectively, obtained using the two approaches were in good agreement. PMID- 19307014 TI - A phase II study of weekly topotecan and docetaxel in heavily treated patients with recurrent uterine and ovarian cancers. AB - OBJECTIVES: A phase II trial designed to evaluate the safety and efficacy of weekly topotecan and docetaxel in heavily treated patients with recurrent uterine or epithelial ovarian cancers. METHODS: Eligible patients with recurrent epithelial ovarian or uterine cancers were treated with weekly topotecan 3.5 mg/m(2) and docetaxel 30 mg/m(2) for 3 consecutive weeks. Cycles were repeated every 4 weeks for 6 cycles or until evidence of disease progression, unacceptable toxicity, or death. Response was assessed as per RECIST or Rustin's criteria. Time to best response and overall survival were calculated using Kaplan-Meier statistical methods. RESULTS: Twenty-seven patients registered, of which 24 were evaluable for response. The majority of patients had received 2 prior chemotherapy regimens. Of the total 86 cycles of chemotherapy that were administered, there were three grade 4 (all neutropenia) and ten grade 3 toxicities. Six of the grade 3 non-hematologic toxicities were unrelated to treatment. There were 8 dose delays and 4 dose reductions. The overall response rate was 25% (95% CI: 7.7%-42.3%, 8% CR, 17% PR), and 38% of the patients had clinical benefit (95% CI: 18.1%-56.9%; CR+PR+13% SD). The median duration of response was 8.5 months (range 3-19 months). The median overall survival was 18.5 months (range 1.8-50.7 months). CONCLUSION: The combination of weekly topotecan and docetaxel has clinical benefit and is well tolerated in this heavily treated patient population. Patients with platinum-resistant tumors had clinical benefit and should be considered for further study with this regimen. PMID- 19307015 TI - Factors associated with deliberate self-harm behaviour among depressed adolescent outpatients. AB - This study examined whether depressed adolescent outpatients with deliberate self harm behaviour (DSH) differed from non-suicidal depressed adolescent outpatients in depressive and anxiety symptoms, alcohol use, perceived social support and number of negative life-events. Depressed adolescent outpatients (n=155) aged 13 19 years were interviewed using K-SADS-PL for DSM-IV Axis I diagnoses and completed self-report questionnaires. Suicidal behaviour was assessed by K-SADS PL suicidality items. Depressed adolescents with DSH were younger, perceived less support from the family, had more severe depressive symptoms and used more alcohol than non-suicidal depressed adolescents. Adolescents with DSH and suicidal ideation or suicide attempts had more depressive and anxiety symptoms than adolescents with DSH only. Adolescents with severe internalizing distress symptoms are at risk not only for DSH, but also additional suicidal behaviour. Family interventions may be needed in the treatment of depressed adolescents with DSH. PMID- 19307016 TI - Influence of marine phytoplankton, transition metals and sunlight on the species distribution of chromium in surface seawater. AB - The photoreduction of Cr(VI) to Cr(III) by marine phytoplankton (diatoms, red and green algae), with or without the presence of transition metals (Fe(III), Cu(II) and Mn(II)) was studied. The direct influence of marine phytoplankton on the photochemical reduction of Cr(VI) was confirmed for the first time, and two kinds of mechanisms were suggested to be responsible for the species transformation: (a) Cr(VI) in excited state could be reduced by the electron donor in its ground state via photo produced electrons; and (b) the solvated electrons reduce the CrO(4)(2-) anions in their ground state. The conversion ratio of Cr(VI) to Cr(III) increased with increasing algae concentration and irradiation time. Different species of marine phytoplankton were found to have different photo reducing abilities. The photochemical redox of transition metals could induce the species transformation of chromium. After photoreduction by marine phytoplankton and transition metals, the ratio of Cr(VI) to Cr(III) was in the range of 1.45 2.16 for five green algae (Tetraselmis levis, Chlorella autotrophica, Dunaliella salina, Nannochloropsis sp., and Tetraselmis subcordiformis), and only 0.48 for Phaeodactylum tricornutum (diatom) and 0.71 for Porphyridium purpureum (red alga). The species distribution of chromium in the sunlit surface seawater was greatly affected by combined effects of marine phytoplankton (main contributor) and transition metals; both synergistic and antagonistic effects were observed. The results provided further insights into the species distribution and the biogeochemical cycle of chromium, and have significant implications for the risk assessment of chromium in the sunlit surface seawater. PMID- 19307017 TI - Rhinocerebral zygomycosis and subsequent treatment decisions in a young patient with AML. PMID- 19307018 TI - The metabolic consequences of imatinib mesylate: Changes on glucose, lypidic and bone metabolism. AB - The therapeutic efficacy of imatinib mesylate is based on its specific inhibition of several tyrosine kinases (TKs) implicated in the disease pathogenesis. These enzymes include BCR/ABL in patients with chronic myeloid leukaemia, PDGF-R alpha and beta in patients with certain myeloproliferative disorders and dermatofibrosarcoma protuberans and c-KIT in patients with gastrointestinal tumors. Most patients tolerate the drug well and apparently no metabolic abnormalities are evidenced during treatment. However, different metabolic effects have been reported as a consequence of imatinib inhibition during treatment of patients with CML. The aim of this review is to report the changes caused by imatinib on glucose, lypidic and bone metabolism. PMID- 19307019 TI - Synthetic peptide issued from Hap1/LipL32 for new early serodiagnosis of human leptospirosis. AB - Leptospirosis is a worldwide zoonosis. Today, serological diagnosis is generally assessed by MAT. We performed ELISA with a synthetic peptide derived from Hap1/lipL32 which is a protein expressed only by pathogenic Leptospira. Repeatability and thresholds were defined with 85 controls sera and 119 hospitalized leptospirosis. The PP-ELISA repeatability and IgM/IgG cut-off values were based on control sera. For these cut-off values, we observed the IgM-PP ELISA specificity of 89%, whereas it was 100% for the IgG. Then, we compared PP ELISA and standard MAT results for leptospirosis patients. The concordance rate for IgM-PP-ELISA and MAT was low (43%), whereas it was 85% for IgG-PP-ELISA and MAT. During the first 5 days after hospitalization, PP-ELISA gave positive results in 13 out of 16 patients (81%) whereas 8 out of 14 patients (57%) were positive to MAT. ELISA using Hap1/lipL 32-derived synthetic peptide PP is an earlier serological diagnosis of human leptospirosis than MAT. PMID- 19307020 TI - Prediction of SVOC vegetation and atmospheric concentrations using calculated deposition velocities. AB - A simple modeling concept was previously applied to study air-foliage exchange of both polybrominated diphenyl ethers (PBDEs) and polycyclic aromatic hydrocarbons (PAHs) using field measurements. In this paper, the predictive capabilities of this framework were tested using an independent set of data collected at the same sampling location for a different time period. Firstly, atmospheric (particle bound and gaseous phase) PBDE and PAH concentrations were used to predict their accumulation in spruce needles. Conversely, we used PBDE and PAH concentrations in vegetation to predict their atmospheric concentrations. In both cases, calculated and measured values showed good agreement, providing evidence that the developed framework and our derived deposition velocities can be used as a predictive tool for these two different classes of semi-volatile organic compounds. PMID- 19307021 TI - Mannan-binding lectin-associated serine protease-2 (MASP-2) in a large cohort of neonates and its clinical associations. AB - One collectin (mannan-binding lectin, MBL) and three ficolins (M-ficolin/ficolin 1, L-ficolin/ficolin-2 and H-ficolin/ficolin-3) share the capability to activate complement via the lectin pathway. This property depends on the ability of these lectins to form complexes with MBL-associated serine proteases (MASPs), particularly MASP-2. We report the results of an investigation of cord blood MASP 2 concentrations in a large, ethnically homogeneous cohort (n=1788) of neonates. The median value of MASP-2 in cord sera was determined to be 93 ng/ml (range <25 812). Serum MASP-2 concentrations correlated with gestational age and birthweight and were significantly lower in premature babies and other pre-term babies compared with term babies. Neonates with MASP-2 concentrations below 42 ng/ml were deemed to be MASP-2 deficient. That group had a shorter mean gestational age and a higher incidence of premature and low birthweight babies, but not of perinatal infections when compared with the others. Indeed, there was a trend towards higher MASP-2 concentrations amongst babies with infections. Among 362 samples tested for the D120G single nucleotide polymorphism (SNP) of the MASP2 gene, no homozygote for that mutation was found. Heterozygosity for this allele significantly influenced the protein concentration, but not the lectin pathway of complement activity (MBL-MASP-2 complex activity). Moreover, no association of this SNP was apparent with prematurity, low birthweight or perinatal infections. PMID- 19307022 TI - Structure-specific binding of [Co(phen)(2)(HPIP)](3+) to a DNA duplex containing sheared G:A mismatch base pairs. AB - The binding of a Co(III) complex to the decanucleotide d(CCGAATGAGG)(2) containing two pairs of G:A mismatches was studied by 2D-NMR, UV absorption, and molecular modeling. NMR investigations indicate that racemic [Co(phen)(2)(HPIP)]Cl(3) [HPIP=2-(2-hydroxyphenyl) imidazo [4,5-f][1,10] phenanthroline] binds the decanucleotide by intercalation: the HPIP ligand selectively inserts between the stacked bases from the minor groove at the terminal regions and from the major groove at the sheared region. Further, molecular modeling revealed that the recognition shows strong enantioselectivity: the Lambda-isomer preferentially intercalates into the T(6)G(7):A(5)A(4) region from the DNA major groove, while Delta-isomer favors the terminal C(1)C(2):G(10)G(9) region and intercalates from the minor groove. Detailed energy analysis suggests that the steric interaction, especially the electrostatic effect, is the primary determinants of the recognition event. Melting experiments indicate that binding stabilizes the DNA duplex and increases the melting temperature by 9.5 degrees C. The intrinsic binding constant of the complex to the mismatched duplex was determined to be 3.5x105M(-1). PMID- 19307023 TI - Dengue virus induces thrombomodulin expression in human endothelial cells and monocytes in vitro. AB - OBJECTIVES: Dengue virus (DV) infections can cause severe life-threatening dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). However, the mechanism to cause hemorrhage in DV infections remains poorly understood. Thrombomodulin (TM), expressed on the surface of endothelial cells and monocytes, is very important in regulation of coagulation and inflammation. Therefore, the effect of DV on the TM expression was studied in vitro using both endothelial cells and monocytes. METHODS AND RESULTS: The expression of TM in human endothelial cell line, HMEC-1, monocytic cell line THP-1 and peripheral blood mononuclear cells derived from human blood was increased after DV infection, UV-inactivated DV or recombinant DV envelop protein domain III stimulation as demonstrated by flow cytometry and immunofluorescent staining. Western blot analysis further confirmed only DV but not enterovirus 71 infection of HMEC-1 cells increased TM protein expression. In addition, RT-PCR analysis showed the increase of TM mRNA as well as other protein C activation-related molecules in DV stimulated HMEC-1 in a dose-dependent manner. CONCLUSION: These results suggest that DV stimulation of human endothelial cells and monocytes can increase the expression of TM, which may contribute to the anticoagulant properties of cells during DV infection. PMID- 19307024 TI - Immune system alterations in sporadic amyotrophic lateral sclerosis patients suggest an ongoing neuroinflammatory process. AB - In this work we show that patients with sporadic amyotrophic lateral sclerosis exhibit immunological alterations in their blood, with respect to healthy controls, such as: i) increased levels of CD4+ cells and decreased levels of CD8+ T lymphocytes, the latter due to the reduced expression of the anti-apoptotic molecule Bcl-2; ii) significantly reduced CD4+CD25+ regulatory T (Treg) cells and monocytes (CD14+) levels in patients at a less severe stage of disease, suggesting their early recruitment towards the CNS area of primary neurodegeneration; iii) reduced expression of HLA-DR and CCR2 expression, as markers of activation, in monocytes. Since resident microglia partially derives from circulating activated monocytes and Treg cells are known to interact with the local microglia, this study strengthens the hypothesis of an involvement of the adaptive immune system associated with a neuroinflammatory process in the pathobiology of ALS. PMID- 19307026 TI - The accuracy of otomicroscopy for the diagnosis of paediatric middle ear effusions. AB - OBJECTIVE: To determine the diagnostic accuracy of otomicroscopy performed by otolaryngology specialists for the diagnosis of paediatric middle ear effusions. METHODS: Prospective study of consecutive children receiving tympanostomy tubes between December 2004 and February 2005 within the Department of Otolaryngology Head & Neck Surgery, Manukau Surgery Centre, Middlemore Hospital, New Zealand. Eighty-six children were included in the study. Intraoperative otomicroscopic examination of the ears was performed under general anaesthesia by two otolaryngology specialists. The presence or absence of middle ear effusion was predicted prior to myringotomy. Intraoperative myringotomy findings were used as the diagnostic reference standard. RESULTS: The prevalence of middle ear effusions was 52.6% (71/135 ears). Otomicroscopy had a sensitivity 94.4% (95% CI: 85.5-98.2), and specificity 93.8% (95% CI: 84.0-98.0). The positive predictive and negative predictive values of otomicroscopy for the identification of middle ear effusions were 94.4% (95% CI: 85.5-98.2), and 93.8% (95% CI: 84.0-98.0) respectively. The overall accuracy of otomicroscopy was 94.1%. There was an excellent level of agreement between otomicroscopy and myringotomy findings (kappa=0.88, 95% CI: 0.80-0.96). CONCLUSIONS: Otomicroscopy performed by the specialist otolaryngologist is an accurate tool for the diagnosis of middle ear effusions. Otomicroscopy performed on the anaesthetised child achieves greater diagnostic accuracy than tympanometry and pneumatic otoscopy. PMID- 19307027 TI - Lack of association between common polymorphisms of epidermal growth factor receptors and nonsyndromic cleft lip with or without cleft palate. AB - OBJECTIVES: Nonsyndromic cleft lip with or without cleft palate (CL/P) is a frequent craniofacial malformation with a complex aetiology. Since the first report of an association between DNA sequence variants at the transforming growth factor alpha gene (TGFA) and nonsyndromic oral clefts, several studies have been carried out, which have produced conflicting results. Overall, TGFA is considered as a genetic clefting modifier in humans. Murine models indicate that the Tgfa product (tgfalpha), as well as its receptor (Egfr), actively participates in palate development. Notably, Egfr null mice showed an increased incidence in orofacial clefts. In the present study, genes which code for subunits of epidermal growth factor receptors (EGFRs) have been considered as candidate genes for CL/P. METHODS: A family based investigation was performed using a sample of 239 case/parent triads. The aim was to test for an allelic association between common non-synonymous polymorphisms in EGFR genes and CL/P. RESULTS AND CONCLUSION: The results did not suggest any evidence of a link between the investigated polymorphisms and CL/P, however the involvement of different polymorphisms or mutations in such genes cannot be excluded. PMID- 19307028 TI - Drug development and the cellular quality control system. AB - Proteins serve in cellular roles that necessitate structural precision, a requirement overseen by the cellular quality control system (QCS). By rejecting misfolded proteins, the QCS protects against aberrant activity. Misfolding and subsequent retention by the QCS results in proteins that might maintain function but become misrouted and cause disease. Correcting the misrouting of misfolded mutant proteins often restores activity and addresses the underlying disease. Because of its small size, the gonadotropin-releasing hormone receptor has been an excellent model for G-protein-coupled receptor trafficking and has recently enabled elucidation of both the requirements to pass the QCS and the biochemical mechanism of rescue by pharmacological chaperones; this information will now enable rational design of these therapeutic agents. Here, we summarize what is known about the relation between receptor structure and interactions with the QCS with a view toward therapeutic development based on the rescue of misfolded and, consequently, misrouted mutants with drugs. PMID- 19307029 TI - MRI in small brains displaying extensive plasticity. AB - Manganese-enhanced magnetic resonance imaging (ME-MRI), blood oxygen-level dependent functional MRI (BOLD fMRI) and diffusion tensor imaging (DTI) can now be applied to animal species as small as mice or songbirds. These techniques confirmed previous findings but are also beginning to reveal new phenomena that were difficult or impossible to study previously. These imaging techniques will lead to major technical and conceptual advances in systems neurosciences. We illustrate these new developments with studies of the song control and auditory systems in songbirds, a spatially organized neuronal circuitry that mediates the acquisition, production and perception of complex learned vocalizations. This neural system is an outstanding model for studying vocal learning, brain steroid hormone action, brain plasticity and lateralization of brain function. PMID- 19307030 TI - Friends and foes in synaptic transmission: the role of tomosyn in vesicle priming. AB - Priming is the process by which vesicles become available for fusion at nerve terminals and is modulated by numerous proteins and second messengers. One of the prominent members of this diverse family is tomosyn. Tomosyn has been identified as a syntaxin-binding protein; it inhibits vesicle priming, but its mode of action is not fully understood. The inhibitory activity of tomosyn depends on its N-terminal WD40-repeat domain and is regulated by the binding of its SNARE motif to syntaxin. Here, we describe new physiological information on the function of tomosyn and address possible interpretations of these results in the framework of the recently described crystal structure of the yeast tomosyn homolog Sro7. We also present possible molecular scenarios for vesicle priming and the involvement of tomosyn in these processes. PMID- 19307032 TI - Detection of the anti-androgenic effect of endocrine disrupting environmental contaminants using in vivo and in vitro assays in the three-spined stickleback. AB - We have previously developed a novel in vitro assay that utilises cultures of primed female stickleback kidney cells for the screening of potential androgenic and anti-androgenic environmental contaminants. Stickleback kidney cells are natural targets for steroid hormones and are able to produce a protein, spiggin, in response to androgenic stimulation. We undertook a combined in vivo/in vitro study where we used the magnitude of spiggin production as an endpoint to test the anti-androgenic properties of the pharmaceutical androgen antagonist flutamide and three environmental contaminants: the organophosphate insecticide fenitrothion, the urea-based herbicide linuron and the fungicide vinclozolin. In vitro, kidney cells were exposed to a range of concentrations [from 10(-14) M (2.5 pg/L) up to 10(-6) M (280 microg/L)] of the test compounds alone for determining agonist activities, or together with 10(-8) M (3 microg/L) dihydrotestosterone (DHT) for determining antagonist activities. An in vivo flow through aquarium-based study was carried out in parallel. Female sticklebacks were exposed to a range of concentrations of the same chemicals alone or in combination with DHT (5 microg/L) for 21 days. All of the compounds significantly inhibited DHT-induced spiggin production in a concentration-dependent manner in both the in vitro (FN > or = FL > or = LN > VZ) and in vivo (FN > FL > or = VZ > LN) assays. Fenitrothion and flutamide inhibited spiggin production in vitro at a concentration as low as 10(-12) M (P < 0.05), while linuron and vinclozolin inhibited DHT-induced spiggin production at concentrations of 10(-10) M (P < 0.05) and 10(-6) M (P < 0.001) respectively. Similarly, fenitrothion and flutamide were the most potent chemicals in vivo and significantly reduced DHT induced spiggin production at a concentration of 10 microg/L and 25 microg/L respectively (P < 0.01). Both linuron and vinclozolin induced a significant decrease in DHT-induced spiggin production at a concentration of 100 microg/L when tested in vivo. In addition, kidney cell primary culture was used to test the (anti-)androgenic effects of the major environmental contaminants: oestradiol (E2), nonylphenol (NP) and bisphenol A (BPA) for the first time in teleosts. We observed that these compounds were able to significantly inhibit spiggin production at high doses (E2: 270 microg/L; NP: 2.2 microg/L; BPA: 2.3 microg/L). When tested in the absence of DHT, none of the compounds showed a significant agonistic activity in either in vivo or in vitro assays. Overall, our data further demonstrate that kidney cell primary culture is a reliable and a sensitive screening tool for the detection of (anti-)androgenic compounds. In addition, our study represents the first attempt to develop a combined in vivo/in vitro screening strategy for assessing the effects of (anti-)androgenic endocrine disrupters. PMID- 19307031 TI - Calcium homeostasis, selective vulnerability and Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disorder of which the core motor symptoms are attributable to the degeneration of dopamine (DA) neurons in the substantia nigra pars compacta (SNc). Recent work has revealed that the engagement of L-type Ca(2+) channels during autonomous pacemaking renders SNc DA neurons susceptible to mitochondrial toxins used to create animal models of PD, indicating that homeostatic Ca(2+) stress could be a determinant of their selective vulnerability. This view is buttressed by the central role of mitochondria and the endoplasmic reticulum (linchpins of current theories about the origins of PD) in Ca(2+) homeostasis. Here, we summarize this evidence and suggest the dual roles had by these organelles could compromise their function, leading to accelerated aging of SNc DA neurons, particularly in the face of genetic or environmental stress. We conclude with a discussion of potential therapeutic strategies for slowing the progression of PD. PMID- 19307033 TI - Up-regulating PPAR-gamma expression and NO concentration, and down-regulating PAI 1 concentration in a rabbit atherosclerotic model: the possible antiatherogenic and antithrombotic effects of atorvastatin. AB - We investigated the effect of atorvastatin on the plasma concentration of plasminogen activator inhibitor-1 (PAI-1) and nitric oxide (NO) in a rabbit model, and the relationship between these effects and peroxisome proliferator activated receptor gamma (PPAR-gamma). In our experiments, 24 male Japanese rabbits were divided into 3 groups: the high-cholesterol diet group (the high-C group), the high-cholesterol diet plus atorvastatin group (the atorvastatin group), and the normal diet group (the control group). All rabbits were killed after a 16-week feeding. The expression of PPAR-gamma and the plasma concentrations of NO and PAI-1 were evaluated by an immunohistochemical assay while the level of the plasma lipid profile was measured using a commercially available kit. The atorvastatin not only reduces the plasma levels of the total cholesterol (TC) and the low-density lipoprotein cholesterol (LDL-C), but also increases the expression of PPAR-gamma and the concentration of NO in comparison to the control group [16.11 +/- 2.35% vs 7.68 +/- 1.04%; 249.30 +/- 27.90 vs 179.12 +/- 28.51 (MUml/L), p<0.05 respectively]. In addition, the concentration of PAI-1 in the atorvastatin group is lower than that in the control group (0.11 +/- 0.01A vs 0.14 +/- 0.02A, p<0.05). The changes of PAI-1 and NO in the atorvastatin group are in good accordance to that of PPAR-gamma. Results show that atorvastatin significantly up-regulates the expression of nuclear transcription factor, namely PPAR-gamma, and induces the changes of the other two factors, which might provide mechanisms for the antiatherosclerotic and antithrombotic effects of atorvastatin. PMID- 19307034 TI - Long-term follow-up of cancer patients receiving radiotherapy for bone metastases: results from a randomised multicentre trial. AB - BACKGROUND AND PURPOSE: The aim of this study was to compare the need for re irradiation, rates of pathological fractures, and spinal cord compressions in patients randomised to single-fraction radiotherapy (8 Gy x 1) or multiple fraction therapy (3 Gy x 10) and with a long-term follow-up. The underlying hypothesis was that single-fraction and multiple-fraction regimens are equally effective. MATERIAL AND METHODS: The present study reports on the Norwegian sub sample of an international large prospective-randomised multicentre study. One hundred and eighty patients with painful skeletal metastases in four Norwegian hospitals were randomised to receive single-fraction (8 Gy x 1) or multiple fraction (3 Gy x 10) radiotherapy. RESULTS: Patients in the single-fraction arm received significantly more re-irradiations as compared to the multiple-fraction arm (27% versus 9%, p=0.002). There were no significant differences in the rate of pathological fractures (5% versus 5%, p=1.00) or spinal cord compressions (1% versus 4%, p=0.37) between the two treatment groups. CONCLUSION: The present study indicates no difference between radiotherapy with 8 Gy x 1 and 3 Gy x 10 for the majority of patients with painful bone metastases, also in a long-term perspective. Importantly, the patients in this study were followed up until death, and the trial showed no disadvantage for 8 Gy x 1 compared to 3 Gy x 10. Despite the fact that single-fraction treatment will imply an approximately 2.5 fold greater need for re-irradiation, single-fraction treatment is considered more convenient for the patients and more cost-effective for the radiotherapy departments. PMID- 19307035 TI - A molecular mechanism of azoxystrobin resistance in Penicillium digitatum UV mutants and a PCR-based assay for detection of azoxystrobin-resistant strains in packing- or store-house isolates. AB - Sixty-five isolates of Pencillium digitatum (Pers.:Fr) Sacc., a causative agent of green mold of postharvest citrus, were collected from various locations in Zhejiang province in 2000, 2005 and 2006, and assayed for their sensitivity to the quinone outside inhibitor (QoI) fungicide azoxystrobin. The results showed that azoxystrobin is highly effective against P. digitatum, in vitro, and that the effective concentrations resulting in reduction of conidial germination and mycelial growth by 50% (EC(50)) averaged 0.0426 microg/ml and 0.0250 microg/ml, respectively. Twenty-eight azoxystrobin-resistant mutants were obtained by UV mutagenesis and subsequent selection on medium amended with azoxystrobin (12 microg/ml) and salicylhydroxamic acid. All obtained mutants were highly resistant to azoxystrobin and their resistance was genetically stable. Analysis of the cytochrome b gene structure of P. digitatum (Pdcyt b) showed the absence of type I intron in the first hot spot region of mutation. These results indicate that P. digitatum is likely to evolve high levels of resistance to azoxystrobin after its application. Analysis of partial sequences of Pdcyt b from both the azoxystrobin sensitive parental isolate and the 28 azoxystrobin-resistant mutants revealed that a point mutation, which leads to the substitution at code 143 of alanine for glycine (G143A), is responsible for the observed azoxystrobin resistance in the laboratory mutants. Based on this point mutation, two allele-specific PCR primers were designed and optimized for allele-specific PCR detection of azoxystrobin resistant isolates of P. digitatum. PMID- 19307036 TI - Microbial, sensory and volatile changes during the anaerobic cold storage of morcilla de Burgos previously inoculated with Weissella viridescens and Leuconostoc mesenteroides. AB - Blood sausage, a widely consumed traditional product, would benefit from an increased commercial life. It is therefore pertinent to investigate the type, the evolution, and the behaviour of the Lactic Acid Bacteria responsible for their spoilage. This study aims to clarify the role played by Weissella viridescens and Leuconostoc mesenteroides, identified as their principal spoilage agents in vacuum-packaged morcilla de Burgos, through the study of microbiological, sensory, and volatile profile changes, following inoculation of the morcilla, both jointly and separately, with the two species. L. mesenteroides grew more rapidly and influenced the drop in pH, milky exudates and the sour smell, whereas W. viridescens influenced vacuum loss. With respect to volatile profiles, L. mesenteroides samples were richer in aldehydes (hexanal) and acids (acetic), on the contrary W. viridescens samples showed greater amounts of alcohols (ethanol) and ketones (acetoin and diacetyl). Both species inoculated together increased particular signs of morcilla spoilage. PMID- 19307037 TI - Administration of a new diabetes-specific enteral formula results in an improved 24h glucose profile in type 2 diabetic patients. AB - AIMS: Study the effect of several boluses of a new diabetes-specific formula (DSF) during the day on 24h glucose profile. METHODS: In this randomized, controlled, double-blind, cross-over study 12 ambulatory type 2 diabetic patients were included. Subjects received a new DSF and an isocaloric standard fibre containing formula (SF) while continuing their anti-diabetic medication. Subjects received 100% of their calculated daily energy requirements as bolus feeding every 3h (5 times/day, starting at 8.00 a.m.+/-1h). RESULTS: Glucose profiles were significantly better after administration of DSF compared with SF determined as mean glucose concentration (+/-SEM) (8.7+/-0.5 versus 9.6+/-0.6 mmol/L, p<0.05 during 24h; 9.4+/-0.6 versus 10.7+/-0.6 mmol/L, p<0.001 during daytime) or as incremental area under the curve during daytime (-44%; p<0.05). Subjects receiving DSF experienced less hyperglycaemic time over 24h (-26%; p<0.05) and during daytime (-30%; p<0.05). Furthermore, lower individual and mean (delta) peak glucose levels were found (p<0.05). No clinically relevant differences in gastrointestinal tolerance were observed. CONCLUSIONS: Using DSF resulted in significantly better 24h and postprandial glucose profiles than fibre-containing SF after bolus administration and may therefore help to improve glycaemic control in diabetic patients. PMID- 19307038 TI - Localization determines function: N-terminally truncated NS5A fragments accumulate in the nucleus and impair HCV replication. AB - BACKGROUND/AIMS: The Hepatitis C Virus (HCV) nonstructural protein 5A (NS5A) is an essential part of the ER-localized HCV-replicon complex. Although NS5A harbours a conserved NLS in its C-terminal domain, NS5A is associated with the cytoplasmic face of the ER by an amphipathic helix close to its N-terminus. METHODS: Intracellular distribution of NS5A in HCV replicating cells was analyzed by confocal microscopy and subcellular fractionation. The effect on HCV replication was analyzed using the JFH-1-based infection/replication system. RESULTS: During viral life cycle N-terminally truncated NS5A fragments are caspase-dependent formed that lack the ER-attachment signal and are localized within the nucleus. These N-terminally truncated fragments inhibit HCV replication. If their formation is blocked by inhibition of caspases HCV replication is increased. The C-terminal domain of NS5A binds to c-Raf and thereby localizes it to the replicon complex. This interaction is essential for HCV replication. The N-terminally truncated NS5A fragments are still able to bind c-Raf. However, due to their nuclear localization they withdraw c-Raf from the replicon complex into the nucleus resulting in an impaired HCV replication. CONCLUSIONS: Formation of N-terminally truncated NS5A fragments could represent a mechanism to regulate HCV replication by withdrawal of essential factors from the replicon complex. PMID- 19307039 TI - Various autoantibodies are found in small-cell lung cancer. PMID- 19307040 TI - Gene tree discordance, phylogenetic inference and the multispecies coalescent. AB - The field of phylogenetics is entering a new era in which trees of historical relationships between species are increasingly inferred from multilocus and genomic data. A major challenge for incorporating such large amounts of data into inference of species trees is that conflicting genealogical histories often exist in different genes throughout the genome. Recent advances in genealogical modeling suggest that resolving close species relationships is not quite as simple as applying more data to the problem. Here we discuss the complexities of genealogical discordance and review the issues that new methods for multilocus species tree inference will need to address to account successfully for naturally occurring genomic variability in evolutionary histories. PMID- 19307041 TI - Understanding the egalitarian revolution in human social evolution. AB - Humans are unique among animals in cooperating in large groups of unrelated individuals, with a high degree of resource sharing. These features challenge traditional evolutionary theories built on kin selection or reciprocity. A recent theoretical model by Gavrilets and colleagues takes a fresh look at the 'egalitarian revolution' that separates humans from our closest relatives, the great apes. The model suggests that information from within-group conflicts leads to the emergence of cooperative alliances and social networks. PMID- 19307042 TI - The honeybee waggle dance: can we follow the steps? AB - The honeybee (Apis mellifera) waggle dance, whereby dancing bees communicate the location of profitable food sources to other bees in the hive, is one of the most celebrated communication behaviours in the animal world. Dance followers, however, often appear to ignore this location information, the so-called dance language, after leaving the nest. Here we consider why foragers follow dances and discuss the function of the dance as a multicomponent signal. We argue that the 'dance language' is just one information component of the waggle dance and that the two terms should not be used synonymously. We propose that location information is often backup information to be used in case information acquired directly from the environment about food sources is not available, is of poor quality or is associated with unprofitable food sources. PMID- 19307043 TI - Intralocus sexual conflict. AB - Intralocus sexual conflict occurs when selection on a shared trait in one sex displaces the other sex from its phenotypic optimum. It arises because many shared traits have a common genetic basis but undergo contrasting selection in the sexes. A recent surge of interest in this evolutionary tug of war has yielded evidence of such conflicts in laboratory and natural populations. Here we highlight outstanding questions about the causes and consequences of intralocus sexual conflict at the genomic level, and its long-term implications for sexual coevolution. Whereas recent thinking has focussed on the role of intralocus sexual conflict as a brake on sexual coevolution, we urge a broader appraisal that also takes account of its potential to drive adaptive evolution and speciation. PMID- 19307044 TI - Ultraviolet leaf pigments as components of autumn colours: a constructive comment on Archetti et al. PMID- 19307045 TI - A gene encoding SMALL ACIDIC PROTEIN 2 potentially mediates the response to synthetic auxin, 2,4-dichlorophenoxyacetic acid, in Arabidopsis thaliana. AB - The SMALL ACIDIC PROTEIN 2 (SMAP2) gene is a paralogue of the SMAP1 gene that mediates the response to the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4 D) in the root of Arabidopsis thaliana. Their encoded proteins, SMAP1 and SMAP2, are similar in calculated molecular weight and isoelectric point, and in having a highly conserved phenylalanine and aspartic acid-rich domain. RNA expression analysis showed that SMAP1 mRNA is present throughout the plant body while SMAP2 mRNA is restricted to siliques and anthers. Over-expression of the SMAP2 gene, as well as SMAP1, by 35S cauliflower mosaic virus promoter restored sensitivity to 2,4-D in the 2,4-D-resistant mutant, aar1, which is defective in SMAP1 function. The results suggest that SMAP2 has an ability to mediate the 2,4-D response and is expressed only in restricted tissues. PMID- 19307046 TI - Cloning and functional analysis of 9-cis-epoxycarotenoid dioxygenase (NCED) genes encoding a key enzyme during abscisic acid biosynthesis from peach and grape fruits. AB - Ripening and senescence are generally controlled by ethylene in climacteric fruits like peaches, and the ripening process of grape, a non-climacteric fruit, may have some relationship to abscisic acid (ABA) function. In order to better understand the role of ABA in ripening and senescence of these two types of fruits, we cloned the 9-cis-epoxycarotenoid dioxygenase (NCED) gene that encodes a key enzyme in ABA biosynthesis from peaches and grapes using an RT-PCR approach. The NCED gene fragments were cloned from peaches (PpNCED1and PpNCED2, each 740bp) and grapes (VVNCED1, 741bp) using degenerate primers designed based on the conserved amino acids sequence of NCEDs in other plants. PpNCED1 showed 78.54% homology with PpNCED2, 74.90% homology with VVNCED1, and both showed high homology to NCEDs from other plants. The expression patterns of PpNCED1 and VVNCED1 were very similar. Both were highly expressed at the beginning of ripening when ABA content becomes high. The maximum ABA preceded ethylene production in peach fruit. ABA in the grape gradually increased from the beginning of ripening and reached the highest level at 20d before the harvest stage. However, ethylene remained at low levels during the entire process of fruit development, including ripening and senescence. ABA content, and ripening and softening of both types of fruits, were promoted or delayed by exogenous ABA or Fluridone (or NDGA) treatment. The roles of ABA and ethylene in the later ripening of fruit are complex. Based on results obtained in this study, we concluded that PpNCED1 and VVNCED1 initiate ABA biosynthesis at the beginning of fruit ripening, and that ABA accumulation might play a key role in the regulation of ripeness and senescence of both peach and grape fruits. PMID- 19307047 TI - [Olanzapine induced rhabdomyolysis and serum creatine kinase increase]. AB - INTRODUCTION: "Atypical" antipsychotics tend to replace traditional antipsychotics as first line therapy for psychotic disorders, due to their better side-effect profile with fewer extrapyramidal manifestations, allowing a better observance. Nevertheless, second-generation antipsychotics may also lead to adverse events such as metabolic disorders, agranulocytosis or muscle damage. Cases of rhabdomyolysis (aside neuroleptic malignant syndrome) have been reported in patients receiving olanzapine (Zyprexa). METHODS: We reviewed the cases of olanzapine induced rhabdomyolysis reported to the French national database of drug adverse events and retrieved additional cases published in the medical literature. RESULTS: We collected 13 cases from the French pharmacovigilance database and eight additional cases from the literature. Seventeen patients needed hospitalization. Creatine kinase (CK) rate ranged between 413 and 34,500 UI/L. Outcome was favorable in 85% of the cases (17 out of 20 cases) after discontinuation of olanzapine. CONCLUSION: Although rhabdomyolysis is a rare side effect (< 1%) of olanzapine, this adverse event should be evoked when a patient with olanzapine presents with muscle pain, unexplained fatigue or weakness. Prompt dosage of CK should be performed. However, it remains uncertain whether a mild and asymptomatic muscle enzyme increase without any metabolic disorder requires the discontinuation of olanzapine therapy. PMID- 19307048 TI - Biomechanical evaluation of two types of short-stemmed hip prostheses compared to the trust plate prosthesis by three-dimensional measurement of micromotions. AB - BACKGROUND: Stemless and short-stemmed hip prostheses have been developed to preserve femoral bone stock. While all these prostheses claim a more or less physiological load transfer, clinical long-term results are only available for the stemless thrust plate prosthesis. In this study, the in vitro primary stability of the thrust plate prosthesis was compared to two types of short stemmed prostheses. In addition to the well-established Mayo prosthesis, the modular Metha prosthesis was tested using cone adapters with 130 degrees and 140 degrees neck-shaft-angles. METHODS: The prostheses were implanted in composite femurs and loaded dynamically (300-1700 N). Three-dimensional micromotions at the bone-prosthesis interface were measured. In addition, the three-dimensional deformations at the surface of the composite femur were measured to gain data on the strain distribution. FINDINGS: For all tested prostheses, the micromotions did not exceed 150 microm, the critical value for osteointegration. The thrust plate prosthesis revealed similar motions as the short-stemmed prostheses. The short-stemmed prosthesis with the 130 degrees cone tended to have the highest micromotions of all tested short-stemmed prostheses. The thrust plate prosthesis revealed the lowest alteration of bone surface deformation after implantation. INTERPRETATION: The comparably low micromotions of the thrust plate prosthesis and the short-stemmed prostheses should be conducive to osseous integration. The higher alteration of load transmission after implantation reveals a higher risk of stress shielding for the short-stemmed prostheses. PMID- 19307049 TI - N400 deficits from semantic matching of pictures in probands and first-degree relatives from multiplex schizophrenia families. AB - Endophenotypes is one emerging strategy in schizophrenia research that is being used to identify the functional importance of genetically transmitted, brain based deficits present in this disease. Currently, event-related potentials (ERPs) are timely used in this search. Several ERPs, including N400, present deficits in relation to schizophrenia. In order to assess the genetic liability of N400 as a possible endophenotype, a picture semantic matching task (congruent and incongruent pairs of pictures) was performed by 21 unaffected first-degree relatives of patients with schizophrenia, 21 DSM-IV diagnosed schizophrenia probands, and 21 control subjects, matched by age, gender and educational level. Probands and relatives were selected form Multiplex schizophrenia families. Significantly reduced N400 amplitude for congruent categories in N400 was found in probands and relatives in relation to controls. The latency onset and the maximum peak latency of N400 were delayed in both, relatives and probands groups compared to control. The voltage maps of incongruous-minus-congruous difference indicate a more reduced right restricted negativity in probands and relatives, when compared to a widely extended bilateral negativity in controls. No general differences were found between patients and relatives. These results demonstrate an electrophysiological deficit in semantic match processing in clinically unaffected first-degree relatives of patients with schizophrenia, suggesting a possible use of this marker as endophenotype. PMID- 19307050 TI - Temporal sequence of hemispheric network activation during semantic processing: a functional network connectivity analysis. AB - To explore the temporal sequence of, and the relationship between, the left and right hemispheres (LH and RH) during semantic memory (SM) processing we identified the neural networks involved in the performance of functional MRI semantic object retrieval task (SORT) using group independent component analysis (ICA) in 47 healthy individuals. SORT requires participants to determine whether word pairs describing object features combine to retrieve an object. Functional network connectivity (FNC) was used to assess the correlations between components' time courses. Results showed that semantic left and right hemisphere networks comprise two independent ICA components. The components' time courses were highly correlated and RH activation preceded that of the LH. Moreover, this correlation was significantly stronger in better vs. poorer performers of the SORT. These results indicate an early activation of the RH that is closely followed by activation of the LH, to facilitate performance during word retrieval from SM. PMID- 19307051 TI - Oligomerisation and thermal stability of polyvalent integrin alpha5beta1 ligands. AB - Synthetic oligomeric integrin alpha5beta1 ligands, specifically immobilised to surfaces, facilitate increased fibroblast cell spreading compared with that associated with the monomer. These ligands consist of a N-terminal fibronectin domain pair, a spacer and a di-, tri- or tetrameric coiled coil. However, it is not yet clear what effect fusion of the fibronectin domains has on the predicted oligomerisation of the coiled coils. Using analytical ultracentrifugation we show that the predicted tetrameric and trimeric coiled coils facilitate a corresponding ligand oligomerisation with half-dissociation at 0.7 and 0.2 microM, respectively. In contrast, the predicted dimeric coiled coil formed both dimers and trimers. Under non-reducing conditions, the unique C-terminal thiol facilitated inter-oligomer dimerisation of the trimeric species, generating hexameric ligands. Disulphide bonding also increased helical stability during thermal unfolding. The work allows the cellular response to these clustered integrin alpha5beta1 ligands to be more accurately interpreted, and has wider implications with respect to the utility of coiled coils as tools to facilitate protein oligomerisation. PMID- 19307052 TI - Re: Vincent Cloutier, Umberto Capitanio, Laurent Zini, et al. Thirty-day mortality after nephrectomy: clinical implications for informed consent. Eur urol 2009;56:998-1005. PMID- 19307053 TI - Expression of the Breast Cancer Metastasis Suppressor 1 (BRMS1) maintains in vitro chemosensitivity of breast cancer cells. AB - The Breast Cancer Metastasis Suppressor 1 (BRMS1) belongs to an expanding category of proteins called metastasis suppressors that demonstrate in vivo metastasis suppression while still allowing growth of the orthotopic tumor. Since BRMS1 decreases either the expression or function of multiple mediators implicated in resistance to chemotherapy (NF-kappaB, AKT, EGFR), we asked whether breast carcinoma cells expressing BRMS1 could be sensitized upon exposure to commonly used therapeutic agents that inhibit some of these same cellular mediators as BRMS1. In this report, we demonstrate that chemosensitivity of breast cancer cells is preserved in the presence of BRMS1. Further, BRMS1 does not change expression of AKT isoforms or PTEN, implicated in chemoresistance to common drug agents. Overall, our data with two different metastatic breast cancer cell lines indicates that BRMS1 expression status may not interfere with the response to commonly used chemotherapeutic agents in the management of solid tumors such as breast cancer. Since tumor protein expression analysis increasingly guides therapy decisions, our data may be of clinical benefit in disease management including profiling for BRMS1 expression before start of therapy. PMID- 19307054 TI - Decursin and decursinol inhibit VEGF-induced angiogenesis by blocking the activation of extracellular signal-regulated kinase and c-Jun N-terminal kinase. AB - The root of Angelica gigas Nakai contains two major coumarins, which have been previously identified as decursin and decursinol. Decursin has been demonstrated to exhibit potent anti-cancer activity both in vitro and in vivo. In this study, we found that decursin and decursinol at non-cytotoxic doses inhibited the VEGF induced proliferation, migration, and capillary-tube formation of HUVECs. Moreover, decursin and decursinol suppressed microvessel formation on chorioallantoic membranes in fertilized eggs and into mouse Matrigel plugs. The oral administration of decursin and decursinol also reduced VEGF-induced angiogenesis in Matrigel. Furthermore, decursin and decursinol reduced the phosphorylation of ERK and JNK, but not p38 MAPK, in VEGF-stimulated HUVECs. Taken together, our results reveal that decursin and decursinol inhibit VEGF induced angiogenesis by reducing the activation of ERK and JNK in HUVECs, and possess potent in vivo anti-angiogenic activity, coupled with the advantage of oral dosing. Thus, these compounds may have the potential for the treatment of cancers dependent on VEGF-induced vascularization. PMID- 19307055 TI - Tyrosine kinase B receptor and BDNF expression in ovarian cancers - Effect on cell migration, angiogenesis and clinical outcome. AB - In this report, we demonstrated that overexpression of tropomyosin-related kinase B (TrkB) was associated with shorter survival in ovarian cancer patients. Brain derived neurotrophic factor (BDNF), the TrkB ligand, induced activation (phosphorylation) of TrkB in a dose dependent manner. Besides demonstrating the effect of BDNF/TrkB pathway in enhancing cancer cell migration and invasion but inhibiting apoptosis, we also report for the first time that exogenous hepatocyte growth factor induced TrkB expression at both mRNA and protein levels as well as phosphorylation. Our findings suggest that BDNF/TrkB pathway is important in ovarian carcinogenesis and TrkB may be a potential therapeutic target for ovarian cancer. PMID- 19307056 TI - Visible light degradation of Orange II using xCuyOz/TiO2 heterojunctions. AB - Cu(2)O/TiO(2), Cu/Cu(2)O/TiO(2) and Cu/Cu(2)O/CuO/TiO(2) heterojunctions were prepared and studied for their potential application as photocatalysts able to induce high performance under visible light. Orange II was used as a representative dye molecule. The effect of the amount and composition of the photosensitizers toward the activation of TiO(2) was studied. In each case, the global mechanism of Inter Particle Electrons Injection (IPEI) was discussed. The highest photocatalytic activity was observed for the system Cu/Cu(2)O/CuO (MB2 catalyst) under visible light (t(1/2)=24 min, k=159.7 x 10(-3)min(-1)) and for the heterojunction cascade Cu/Cu(2)O/CuO/TiO(2) (MB2 (50%)/TiO(2)) under UV-vis light (t(1/2)=4 min, k=1342 x 10(-3)min(-1)). In the last case, the high performance was attributed firstly to the electromotive forces developed under this configuration in which CuO energy bands mediate the electrons transfer from Cu(2)O to TiO(2). The formation of monobloc sensitizers also accounts for the decrease of the probability of the charges lost. It was demonstrated that "Cu(2)O/CuO" governs the capability of the heterojunction cascade and Cu does not play a significant role regardless of the heterojunction cascade efficiency. The electrical energy consumption per order of magnitude for photocatalytic degradation of Orange II was investigated for some representative catalytic systems. Visible/MB2 and UV/vis MB2 (50%)/TiO(2) exhibited respectively 0.340 and 0.05 kW hm(-3) demonstrating the high efficiency of the systems. PMID- 19307057 TI - A means to estimate thermal and kinetic parameters of coal dust layer from hot surface ignition tests. AB - A method to estimate thermal and kinetic parameters of Pittsburgh seam coal subject to thermal runaway is presented using the standard ASTM E 2021 hot surface ignition test apparatus. Parameters include thermal conductivity (k), activation energy (E), coupled term (QA) of heat of reaction (Q) and pre exponential factor (A) which are required, but rarely known input values to determine the thermal runaway propensity of a dust material. Four different dust layer thicknesses: 6.4, 12.7, 19.1 and 25.4mm, are tested, and among them, a single steady state dust layer temperature profile of 12.7 mm thick dust layer is used to estimate k, E and QA. k is calculated by equating heat flux from the hot surface layer and heat loss rate on the boundary assuming negligible heat generation in the coal dust layer at a low hot surface temperature. E and QA are calculated by optimizing a numerically estimated steady state dust layer temperature distribution to the experimentally obtained temperature profile of a 12.7 mm thick dust layer. Two unknowns, E and QA, are reduced to one from the correlation of E and QA obtained at criticality of thermal runaway. The estimated k is 0.1 W/mK matching the previously reported value. E ranges from 61.7 to 83.1 kJ/mol, and the corresponding QA ranges from 1.7 x 10(9) to 4.8 x 10(11)J/kg s. The mean values of E (72.4 kJ/mol) and QA (2.8 x 10(10)J/kg s) are used to predict the critical hot surface temperatures for other thicknesses, and good agreement is observed between measured and experimental values. Also, the estimated E and QA ranges match the corresponding ranges calculated from the multiple tests method and values reported in previous research. PMID- 19307058 TI - Neonatal pain, parenting stress and interaction, in relation to cognitive and motor development at 8 and 18 months in preterm infants. AB - Procedural pain in the neonatal intensive care unit triggers a cascade of physiological, behavioral and hormonal disruptions which may contribute to altered neurodevelopment in infants born very preterm, who undergo prolonged hospitalization at a time of physiological immaturity and rapid brain development. The aim of this study was to examine relationships between cumulative procedural pain (number of skin-breaking procedures from birth to term, adjusted for early illness severity and overall intravenous morphine exposure), and later cognitive, motor abilities and behavior in very preterm infants at 8 and 18 months corrected chronological age (CCA), and further, to evaluate the extent to which parenting factors modulate these relationships over time. Participants were N=211 infants (n=137 born preterm 32 weeks gestational age [GA] and n=74 full-term controls) followed prospectively since birth. Infants with significant neonatal brain injury (periventricular leucomalacia, grade 3 or 4 intraventricular hemorrhage) and/or major sensori-neural impairments, were excluded. Poorer cognition and motor function were associated with higher number of skin-breaking procedures, independent of early illness severity, overall intravenous morphine, and exposure to postnatal steroids. The number of skin breaking procedures as a marker of neonatal pain was closely related to days on mechanical ventilation. In general, greater overall exposure to intravenous morphine was associated with poorer motor development at 8 months, but not at 18 months CCA, however, specific protocols for morphine administration were not evaluated. Lower parenting stress modulated effects of neonatal pain, only on cognitive outcome at 18 months. PMID- 19307059 TI - Profiling of dynamically changed gene expression in dorsal root ganglia post peripheral nerve injury and a critical role of injury-induced glial fibrillary acidic protein in maintenance of pain behaviors [corrected]. AB - To explore cellular changes in sensory neurons after nerve injury and to identify potential target genes contributing to different stages of neuropathic pain development, we used Affymetrix oligo arrays to profile gene expression patterns in L5/6 dorsal root ganglia (DRG) from the neuropathic pain model of left L5/6 spinal nerve ligation at different stages of neuropathic pain development. Our data indicated that nerve injury induced changes in expression of genes with similar biological functions in a temporal specific manner that correlates with particular stages of neuropathic pain development, indicating dynamic neuroplasticity in the DRG in response to peripheral nerve injury and during neuropathic pain development. Data from post-array validation indicated that there was a temporal correlation between injury-induced expression of the glial fibrillary acidic protein (GFAP), a marker for activated astrocytes, and neuropathic pain development. Spinal nerve ligation injury in GFAP knockout mice resulted in neuropathic pain states with similar onset, but a shortened duration compared with that in age, and gender-matched wild-type littermates. Intrathecal GFAP antisense oligonucleotide treatment in injured rats with neuropathic pain states reversed injury-induced behavioral hypersensitivity and GFAP upregulation in DRG and spinal cord. Together, these findings indicate that injury-induced GFAP upregulation not only serves as a marker for astrocyte activation, but it may also play a critical, but yet identified, role in the maintenance of neuropathic pain states. PMID- 19307060 TI - Role of rat sensory neuron-specific receptor (rSNSR1) in inflammatory pain: contribution of TRPV1 to SNSR signaling in the pain pathway. AB - Sensory neuron-specific receptors (SNSRs) belong to a large family of GPCRs, known as Mrgs (Mas-related genes), many of which are preferentially expressed in primary afferent nociceptors. Selective SNSR agonists produce pain-like behaviors in rats, showing that SNSR activation is sufficient to produce pain. However, it is unknown whether SNSR activation is necessary for pain either in the normal condition or in pathological pain states. Here we used small interfering RNA (siRNA) to acutely knockdown rat SNSR1 and test the hypothesis that this receptor mediates pain responses. Administration of siRNA to the lumbar spinal cord in rats dose-dependently knocked down rSNSR1 mRNA and protein and abolished heat hyperalgesia evoked by intradermal administration of specific rSNSR1 agonists. In rats with levels of rSNSR1 knockdown sufficient to block responses to the SNSR1 agonists, there was no effect on normal pain responses, but there was a significant reduction of heat hyperalgesia in an inflammatory pain model (Complete Freund's Adjuvant), supporting a role for rSNSR1 in inflammatory pain. Further in vivo studies revealed that SNSR1 knockdown had no effect on responses to intradermal capsaicin, a selective TRPV1 agonist. In contrast, a selective TRPV1 antagonist abolished heat hyperalgesia produced by an SNSR agonist, suggesting that TRPV1 receptors mediate rSNSR1-evoked responses. We also found that rSNSR1-like immunoreactivity, like TRPV1, is localized in the superficial dorsal horn of the spinal cord. We propose that rSNSR1 represents a new member of the receptors expressed on chemosensitive nociceptors responsible for detecting the "inflammatory soup" of mediators generated by tissue damage. PMID- 19307061 TI - Does the central nervous system play a role in vitamin D deficiency-related chronic pain? PMID- 19307062 TI - Neuroendocrine and psychometric evaluation of a placebo version of the 'Trier Social Stress Test'. AB - The "Trier Social Stress Test" (TSST) is one of the most prominent laboratory stress paradigms. It is often used to investigate the effects of stress on cognitive or affective parameters. Such studies need a non-stress control condition. However, control conditions currently employed are often rather ill defined and do not parallel important modulating variables, e.g., physical or cognitive load of the TSST. We here introduce a placebo version of the TSST, which contains a free speech and a simple mental arithmetic task without uncontrollability and social-evaluative threat. In two studies, this control condition was evaluated using salivary markers of stress reactivity (cortisol and alpha-amylase) and a questionnaire for anticipatory cognitive stress appraisal (PASA). In experiment 1 participants who were treated with the placebo condition showed no cortisol response and a small, but significant salivary alpha-amylase (sAA) response. Both responses were significantly smaller than those of TSST treated participants. The placebo-treated participants also rated the treatment situation as less stressful. In experiment 2 a crossover study with the use of an intercom to instruct the participants and ensure their compliance was conducted. Again there was a strong cortisol response to the TSST, which differed significantly from the cortisol levels observed during the placebo condition. Importantly the cortisol response was not influenced by treatment order (TSST or placebo first). However, in this study we found similar reactions between TSST- and placebo-treated participants with regard to sAA-response. We suggest that the introduced placebo protocol for the TSST is a promising tool for future psychobiological research. The exact procedure for a given experiment should be tailored to the specific needs of the empirical question studied. PMID- 19307063 TI - Apocrine sweat gland obstruction by antiperspirants allowing transdermal absorption of cutaneous generated hormones and pheromones as a link to the observed incidence rates of breast and prostate cancer in the 20th century. AB - Breast and prostate cancer share similarities and likely represent homologous cancers in females and males, respectively. The role of hormones such as testosterone and estrogen in carcinogenesis is well established. Despite worldwide research efforts, the pathogenesis of these diseases is largely not well understood. Personal care products containing estrogens or xenoestrogens have raised concern as a breast cancer risk, especially in young African-American women. In the United States (US) there is a parallel rise in the incidence in breast and prostate cancer compared to selected non-hormone dependent tumors. Observed US and global breast and prostate cancer incidence increases were occurring before exogenous hormone replacement and xenoestrogen exposure were commonplace. An unintentional, inadvertent, and long term hormone exposure may occur from transdermal absorption of sex hormones and pheromones (androgens) from axillary apocrine sweat gland obstruction by aluminum-based antiperspirants. The global rise in antiperspirant use parallels rises in breast and prostate cancer incidence and mortality rates. A multi-disciplinary literature based set of evidence is presented on how such a link is possible, to prompt confirmatory investigations in the pursuit of unmet needs in breast and prostate cancer etiology and prevention. PMID- 19307064 TI - Melanocyte as a possible key cell in the pathogenesis of psoriasis vulgaris. AB - Current research in pathogenesis of psoriasis vulgaris suggests that the inflammatory mechanisms are immune based and most likely initiated and maintained by T cells. However, the question of lymphocyte being an initiator of psoriatic events remains open so far. Clinical observations such as plaque symmetry, stress induced onset or exacerbations, pruritus, and possibility of generalization, suggest a role of the nervous system and neurogenic inflammation in pathogenesis. A key to understanding the role of melanocyte in psoriasis is their ability to act as regulatory cell in maintaining epidermal homeostasis. In suggested hypothetic event, melanocyte, acting as a local "stress sensor", provide communicatory link between CNS and skin. The disease probably begins with so far unknown signal directed through neuronal network to the melanocyte, placed in the center of epidermal unit. That signal governs keratinocyte cellular activities and lead to reactive abnormal epidermal differentiation and hyperproliferation. Increased proliferation of basal keratinocytes and high metabolic demands creates angiogenesis in papillary dermis and elongation of dermal papillae. Stimulated melanocytes and basal keratinocytes become an important source of proinflammatory cytokines that attract lymphocytes in dermis. In conclusion, according to our hypothesis, lymphocyte infiltrate in psoriasis is secondary event rather than vice versa as presented in the literature. PMID- 19307065 TI - Cystic papillary meningioma with subarachnoid dissemination: a case report and review of the literature. AB - Meningiomas usually present as benign tumors corresponding to WHO grade I. The development of the papillary variant of meningiomas with cyst formation in the central nervous system is extremely rare. We report a case of cystic papillary meningioma in a young female occurring in the lateral ventricle with invasion of brain parenchyma and dissemination of subarachnoid space. The tumor exhibits a marked peritumoral cyst, with contrast enhancement on magnetic resonance imaging (MRI) in accordance with type 2 of Zee's classification of cystic meningioma. Histologically, the tumor displays a classical perivascular pseudopapillary pattern with focal necrosis and subarachnoid space dissemination. Tumor cells are diffusely positive for epithelial membrane antigen (EMA) and vimentin, but lack immunoreactivity for cytokeratin (CK) and glial fibrillary acidic protein (GFAP). MIB-1 labeling is high, accounting for 5% of tumor focally. A diagnosis of primary intraventricular cystic papillary meningioma with subarachnoid space dissemination (WHO grade III) was made. To our knowledge, there is no report describing the radiological and histological characteristics of cystic papillary meningioma presenting in the lateral ventricle. In addition, the biological behavior and the clinical outcome of this tumor are also discussed. PMID- 19307066 TI - Preoperative intensity-modulated radiotherapy combined with temozolomide for locally advanced soft-tissue sarcoma. AB - PURPOSE: To evaluate the toxicity and efficacy of preoperative intensity modulated radiotherapy (IMRT) combined with temozolomide to improve local tumor control in soft-tissue sarcoma (STS). PATIENTS AND METHODS: A cohort of 15 consecutive patients with nonmetastasized, primary high-grade or locally recurrent Stage III (n = 14) or IIb (n = 1) STS not amenable to surgical resection without significant organ or extremity function loss was prospectively investigated. Median tumor size was 9.8 cm, and most tumors were non-extremity sarcomas. Patients preoperatively received 50 mg/m(2) of temozolomide during IMRT (50.4 Gy). Resection was intended 6 weeks thereafter. Toxicity was assessed by the National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0, and response was assessed by Response Evaluation Criteria in Solid Tumors. RESULTS: Of 15 patients, 14 completed preoperative treatment. No Grade 4 toxicities occurred. Nausea and vomiting were the most frequent Grade 3 toxicities. The most frequent toxicities of any grade were dermatologic, gastrointestinal, and hematologic. Response was partial response in 5, stable disease in 7, and progressive disease in 2 patients. Ten patients underwent surgery: 7 were resected with clear margins (R0), and 2 patients had an R1 resection; in 1 patient the tumor was not resectable. Postoperative complications occurred in 4 patients. Five patients did not undergo surgery because of intercurrent metastatic disease, unresectable disease, or refusal. CONCLUSIONS: Preoperative chemoradiation with temozolomide and IMRT can be administered safely and with promising efficacy in patients with locally advanced STS. PMID- 19307067 TI - Induction cisplatin and fluorouracil-based chemotherapy followed by concurrent chemoradiation for muscle-invasive bladder cancer. AB - PURPOSE: To evaluate a multimodality bladder-preserving therapy in patients with muscle-invasive bladder cancer. METHODS AND MATERIALS: Patients with stages T2 4aN0M0 bladder cancer suitable for cystectomy underwent radical transurethral resection and induction chemotherapy, followed by concurrent chemoradiotherapy (CCRT). Patients with a Karnofsky performance status (KPS) <80 or age > or =70 years underwent Protocol A: induction chemotherapy with three cycles of the cisplatin and 5-fluorouracil (CF) regimen, and CCRT with six doses of weekly cisplatin and 64.8 Gy radiotherapy given with the shrinking-field technique. Patients with KPS > or =80 and age <70 years underwent Protocol B: induction chemotherapy with three cycles of weekly paclitaxel plus the CF regimen, and CCRT with six doses of weekly paclitaxel and cisplatin plus 64.8 Gy radiotherapy. Interval cystoscopy was employed after induction chemotherapy and when radiotherapy reached 43.2 Gy. Patients without a complete response (CR) were referred for salvage cystectomy. RESULTS: Among 30 patients (median, 66 years) enrolled, 17 and 13 patients underwent Protocol A and B, respectively. After induction chemotherapy, 23 patients achieved CR. Five (17%) of 7 patients without CR underwent salvage cystectomy. Overall, 28 patients (93%) completed the protocol treatment. Of 22 patients who completed CCRT, 1 had recurrence with carcinoma in situ and 3 had distant metastases. After a median follow-up of 47 months, overall and progression-free survival rate for all patients were 77% and 54% at 3 years, respectively. Of 19 surviving patients, 15 (79%) retained functioning bladders. CONCLUSIONS: Our protocols may be alternatives to cystectomy for selected patients who wish to preserve the bladder. PMID- 19307068 TI - Organizational factors and collaboration and integration activities in criminal justice and drug abuse treatment agencies. AB - Despite strong interest in improving collaborations between correctional and substance abuse treatment organizations, there is a lack of empirical data describing the existing practices. The current study used a national survey of correctional administrators to examine organizational factors related to cross agency collaboration and integration activities between corrections and substance abuse treatment organizations. Using a measure of collaboration that scaled cross agency activities from less structured, informal networking and coordination to more structured and formalized levels of cooperation and collaboration, we found that different correctional settings (e.g., community corrections, jails, prisons) differed significantly in terms of their collaborative activities with substance abuse treatment agencies. We also found that the organizational characteristics that were associated with different levels of collaboration and integration differed across the correctional settings. Further research is needed to better understand how and why correctional agencies decide to formalize collaborative arrangements with treatment agencies and whether these efforts lead to more favorable outcomes. PMID- 19307069 TI - Risk management of drug products and the U.S. Food and Drug Administration: evolution and context. AB - This paper summarizes the background and origins of pharmaceutical risk management and minimization principles and approaches as reflected in FDA statute, policy, and practice. It describes the history of early "risk management" programs, such as the patient package inserts (PPIs) introduced for oral contraceptives in 1976 and medication guides developed for products with safety concerns over the past decade. Exemplary products and programs that include restricted distribution systems such as the early clozapine "blood for drug" program are discussed. The principles and tools described in the US Food and Drug Administration (FDA) risk management guidances of 2005 are likely to be relied upon as the REMS (Risk Evaluation and Mitigation Strategies) mandated by the FDA Amendments Act (FDAAA) of 2007 are implemented. PMID- 19307070 TI - Cardiovascular disease prevention in women: Impact of dietary interventions. AB - In Western countries, cardiovascular disease is the leading cause of death in women. Prevalence of risk factors and clinical presentation are different in men and women and it is important to take these differences into account in both prevention and treatment. A healthy diet can substantially reduce cardiovascular risk. This article summarises the evidence of the effects of dietary interventions in women and discuss gender differences where data are available. For a number of specific dietary factors, there is some evidence of gender specific effects. However, overall risk estimates are not very different from the ones observed in men, and, at this stage, the evidence is insufficient to advocate gender-specific dietary guidelines. Thus a heart-healthy diet is the same for both men and women. However, dietary advice might be more effective in women because of their stronger beliefs in the values of healthy eating. More efforts are needed to further increase awareness of cardiovascular risk and the benefits of a healthy diet. Thus, dietary counselling should be an integral part of the cardiovascular risk management in women. PMID- 19307071 TI - Triggers of apoptosis in the immature brain. AB - Apoptosis occurs physiologically in the mammalian brain during the period of the growth spurt, which in human starts in the 3rd trimester of gestation and ends by the third year of life. Environmental factors can interact with programmed cell death mechanisms to increase the number of neurons undergoing apoptosis and thus produce neuropathological sequelae in the brain. In a series of studies it could be shown that classes of drugs which block N-methyl-D-aspartate (NMDA) glutamate receptors, promote gamma-aminobutyric-acid (GABA(A)) receptor activation or block voltage gated sodium channels, when administered to immature rodents during the period of the brain growth spurt, trigger widespread apoptotic neurodegeneration throughout the developing brain. Studies have also shown that short exposures to non-physiologic oxygen levels can trigger apoptotic neurodegeneration in the brains of infant rodents. Pathomechanisms involved in the proapoptotic action of sedative and anticonvulsant drugs and oxygen include decreased expression of neurotrophins, inactivation of survival signaling proteins, activation of inflammatory cytokines as well as oxidative stress. These findings raise concerns pertaining to the treatment of infants and young children with sedative and anticonvulsant drugs and premature infants with oxygen. The experimental findings imply that new approaches should be developed for patients within these vulnerable age groups. PMID- 19307072 TI - [The French "Haut Conseil de sante publique"]. PMID- 19307073 TI - [Public health surveillance and role of administrative data]. AB - BACKGROUND: Public health surveillance constitutes an important activity since it helps in identifying health needs through data collection, and contributes to decision making and actions by analyzing and interpreting data and communicating key results. METHODS: This paper presents a discussion on the concept of public health surveillance, its objectives and its historical evolution. It deals with the importance of surveillance systems while describing their components and challenges. In addition, the authors point out the importance of administrative data as a relevant source for the surveillance of public health problems, particularly chronic diseases and risk factors. RESULTS: This theoretical discussion leads to the proposal of a conceptual model for surveillance systems, which integrates implementation and evaluation. CONCLUSION: This article provides a summary of the concept of public health surveillance and underlines the general aspects to be considered by the managers of surveillance systems. It also discusses the use of administrative data for surveillance. PMID- 19307074 TI - [Gastric polyps: how to recognize? Which to resect?]. PMID- 19307075 TI - [Duodenal adenomas: diagnostic and treatment]. PMID- 19307076 TI - [PET-CT in gastrointestinal cancer]. PMID- 19307077 TI - An innovative approach to the analysis of 3-[4-(2-methylpropyl)phenyl]propanoic acid as an impurity of ibuprofen on a carbon-coated zirconia stationary phase. AB - 3-[4-(2-Methylpropyl)phenyl]propanoic acid has been introduced as impurity F to the European Pharmacopoeia in its Supplement 4.2. In contrast to other impurities, which are evaluated by HPLC, the content of impurity F is determined by gas chromatography after previous derivatization. Thus a novel reversed-phase HPLC method was developed to simplify the evaluation of pharmacopoeial impurity F of ibuprofen. Favourable properties of zirconia stationary phases were employed for this purpose. The HPLC separation was achieved on a Zr-CARB column (150 mm x 4.6mm i.d., 5 microm) using the mobile phase acetonitrile-phosphate buffer (pH 3.5, 25 mM) (38:62, v/v), temperature 80 degrees C and the flow rate 1.2 ml min( 1). The fluorescence detection was employed to enhance the sensitivity of the method. Optimal detection parameters were chosen on the basis of fluorescence spectra of the analytes. The excitation and emission wavelengths were 220 nm and 285 nm, respectively. The analysis was completed within 25 min. The subsequent validation of the method confirmed the applicability of method for the analytical assay of impurity F. PMID- 19307078 TI - Proliferative capacity of vein graft smooth muscle cells and fibroblasts in vitro correlates with graft stenosis. AB - OBJECTIVE: About a quarter of peripheral vein grafts fail due in part to intimal hyperplasia. The proliferative capacity and response to growth inhibitors of medial smooth muscle cells and adventitial fibroblasts in vitro were studied to test the hypothesis that intrinsic differences in cells of vein grafts are associated with graft failure. METHODS: Cells were grown from explants of the medial and adventitial layers of samples of vein grafts obtained at the time of implantation. Vein graft patency and function were monitored over the first 12 months using ankle pressures and Duplex ultrasound to determine vein graft status. Cells were obtained from veins from 11 patients whose grafts remained patent (non-stenotic) and from seven patients whose grafts developed stenosis. Smooth muscle cells (SMCs) derived from media and fibroblasts derived from adventitia were growth arrested in serum-free medium and then stimulated with 1 muM sphingosine-1-phosphate (S1P), 10 nM thrombin, 10 ng/ml epidermal growth factor (EGF), 10 ng/ml platelet-derived growth factor-BB (PDGF-BB), PDGF-BB plus S1P, or PDGF-BB plus thrombin for determination of incorporation of [(3)H] thymidine into DNA. Cells receiving PDGF-BB or thrombin were also treated with or without 100 microg/ml heparin, which is a growth inhibitor. Cells receiving thrombin were also treated with or without 150 nM AG1478, an EGF receptor kinase inhibitor. RESULTS: SMCs and fibroblasts from veins of patients that developed stenosis responded more to the growth factors, such as PDGF-BB alone or in combination with thrombin or S1P, than cells from veins of patients that remained patent (P = .012). In addition, while PDGF-BB-mediated proliferation of fibroblasts from grafts that remained patent was inhibited by heparin (P < .03), PDGF-BB-mediated proliferation of fibroblasts from veins that developed stenosis was not (P > .5). CONCLUSION: Inherent differences in the proliferative response of vein graft cells to PDGF-BB and heparin may explain, in part, the variability among patients regarding long term patency of vein grafts. PMID- 19307079 TI - True abdominal aortic aneurysm in Marfan syndrome. AB - OBJECTIVES: True abdominal aortic aneurysm (AAA) in patients with Marfan syndrome is relatively rare because most aortic aneurysms in this disease are dissecting aneurysms in the thoracic area. Therefore, its clinical characteristics and long term outcome are still unclear. METHODS: We examined six patients (3 men, 3 women) with Marfan syndrome who had a true AAA. These patients underwent surgical treatment from 1972 to 2004, and we investigated the clinical and histologic findings. RESULTS: The patients were a median age of 45 years (range, 23-73 years) at the time of operation. The median maximum AAA diameter was 76 mm (range, 30-140 mm). Two AAAs ruptured, one of which had twice undergone stent graft insertion before open repair. There was one anastomotic aneurysm and three aortic dissections with additional repair. Marfan-related cardiac complications, all found perioperatively or postoperatively, comprised three patients with annuloaortic ectasia and four with aortic regurgitation. Three patients died, including one death during the operation. Only slight mural thrombus was seen at nonruptured AAAs, and each surgical specimen of aneurysmal wall demonstrated significant cystic medial necrosis in the tunica media. CONCLUSIONS: True AAAs in Marfan syndrome seemed to have several specific features, such as the tendency to occur in relatively young patients, lack of mural thrombus, and susceptibility to dissection and rupture, and the patients have poor life expectancy. Therefore, careful follow-up, keeping these features in mind, is important to treat Marfan syndrome patients with a true AAA. PMID- 19307080 TI - Carotid artery stenting: Impact of practitioner specialty and volume on outcomes and resource utilization. AB - OBJECTIVES: A variety of endovascular specialists perform carotid artery stenting (CAS), but little data exist on outcomes and resource utilization among these specialists. We analyzed differences in outcomes after CAS was performed by radiologists (RAD), cardiologists (CRD), and vascular surgeons (VAS). METHODS: Secondary data analysis of the 2005-2006 State Inpatient Databases for New Jersey were analyzed. Patients with elective admission to the hospital who had CAS procedure or = 20 mm2 in 2 patients), and 15 patients had no MR at rest. Mean exercise-induced change (Delta) in mitral ERO was reduced by CRT (8 +/- 7 mm2 vs 1 +/- 4 mm2, P < .00001). In patients with functional MR at rest, Delta FSV during dynamic exercise was greater with CRT On than CRT Off (4 +/- 8 vs -2 +/- 7 mL, P = .0002), whereas CRT did not significantly affect Delta FSV in patients without MR at rest (9 +/- 9 mL vs 9 +/ 9 mL, P = .93). Similarly, Delta cardiac output was greater with CRT On than CRT Off (1.6 +/- 1.2 L/min vs 1.1 +/- 1.2 L/min, P = .002) in patients with functional MR at rest, whereas Delta cardiac output was similar with CRT On and CRT Off in patients without MR at rest (1.9 +/- 1.4 L/min vs 2.0 +/- 1.2 L/min, P = .59). Severity of functional MR decreased or failed to increase, whereas cardiac output improved during exercise in 9 of 26 patients (34%) with CRT On and in only 2 of 26 patients (8%) with CRT Off (P = .039). CONCLUSION: Functional MR at rest may be an important determinant of the acute hemodynamic response to CRT during exercise. PMID- 19307097 TI - Pseudonormal mitral filling is associated with similarly poor prognosis as restrictive filling in patients with heart failure and coronary heart disease: a systematic review and meta-analysis of prospective studies. AB - OBJECTIVE: The study objective was to more precisely evaluate the link between the pseudonormal mitral filling pattern and death by way of systematic review and meta-analysis. METHODS: Patients with heart failure (HF) and coronary artery disease (CAD) were included. Online databases were searched for prospective studies of patients with HF and CAD who had comprehensive echocardiography. Mortality in patients with pseudonormal filling was compared with restrictive filling and other nonrestrictive filling patterns, including normal and abnormal relaxation. Review Manager Version 4.2.7 software was used for the analysis. RESULTS: Seven studies (5 HF and 2 CAD) were identified, and 887 patients (244 deaths) were included. The pseudonormal filling pattern conferred a 4-fold increase in odds of death compared with abnormal relaxation/normal (odds ratio 4.46; 95% confidence interval, 2.87-6.92). Outcome was similar when restrictive filling was compared with pseudonormal filling (odds ratio 1.16; 95% confidence interval, 0.78-1.74). Death was the main outcome measure. CONCLUSION: This literature-based meta-analysis, pooling results from 7 prospective studies, demonstrates the 4-fold increase in odds of death associated with pseudonormal filling compared with abnormal relaxation/normal. The pseudonormal filling pattern and restrictive filling pattern are associated with similar risk of death. These data further support the need for a comprehensive assessment of diastolic filling, including assessment for pseudonormal filling, as part of routine echocardiographic risk stratification in patients with HF and CAD. PMID- 19307098 TI - A novel non-invasive method of estimating pulmonary vascular resistance in patients with pulmonary arterial hypertension. AB - BACKGROUND: The assessment of pulmonary vascular resistance (PVR) plays an important role in the diagnosis and management of pulmonary arterial hypertension (PAH). The main objective of this study was to determine whether the noninvasive index of systolic pulmonary arterial pressure (SPAP) to heart rate (HR) times the right ventricular outflow tract time-velocity integral (TVI(RVOT)) (SPAP/[HR x TVI(RVOT)]) provides clinically useful estimations of PVR in PAH. METHODS: Doppler echocardiography and right-heart catheterization were performed in 51 consecutive patients with established PAH. The ratio of SPAP/(HR x TVI(RVOT)) was then correlated with invasive indexed PVR (PVRI) using regression and Bland Altman analysis. Using receiver operating characteristic curve analysis, a cutoff value for the Doppler equation was generated to identify patients with PVRI > or = 15 Wood units (WU)/m2. RESULTS: The mean pulmonary arterial pressure was 52 +/- 15 mm Hg, the mean cardiac index was 2.2 +/- 0.6 L/min/m2, and the mean PVRI was 20.5 +/- 9.6 WU/m2. The ratio of SPAP/(HR x TVI(RVOT)) correlated very well with invasive PVRI measurements (r = 0.860; 95% confidence interval, 0.759-0.920). A cutoff value of 0.076 provided well-balanced sensitivity (86%) and specificity (82%) to determine PVRI > 15 WU/m2. A cutoff value of 0.057 increased sensitivity to 97% and decreased specificity to 65%. CONCLUSION: The novel index of SPAP/(HR x TVI(RVOT)) provides useful estimations of PVRI in patients with PAH. PMID- 19307099 TI - Comparative diagnostic accuracy of multiplane and multislice three-dimensional dobutamine stress echocardiography in the diagnosis of coronary artery disease. AB - BACKGROUND: Although real-time three-dimensional echocardiography (RT3DE) is useful for delineating the extent and severity of stress-induced wall motion abnormalities during dobutamine stress echocardiography (DSE), it also provides simultaneous multiple two-dimensional cut planes, which may potentially improve the detection of stress-induced wall motion abnormalities. The aim of this study was to determine the comparative diagnostic accuracy of RT3DE in multiplane and multislice modes for the diagnosis of coronary artery disease (CAD) during DSE against coronary angiography reference. METHODS: Multiplane and multislice cut planes (3 V, GE Healthcare, Milwaukee, WI) at rest and peak dose of dobutamine were acquired in 71 patients with known or suspected CAD. In multiplane mode, matrix array transducer allowed the simultaneous visualization of parasternal long and short-axis views or apical 4-, 2-, and 3-chamber views. From full-volume datasets, 9 equidistant 2-dimensional short-axis images from LV base to apex were extracted and simultaneously displayed (multislice mode). Visual assessment of regional wall motion was performed. Coronary angiography was performed within 48 hours and used as a reference. RESULTS: Abnormal findings (new or worsened wall motion abnormalities or fixed wall motion abnormalities) were noted in 36 patients by multiplane mode and 28 patients by multislice mode. Coronary angiography showed significant stenosis in 32 of 71 patients in 49 of 213 coronary arteries. On a patient basis, sensitivity was not different, but specificity was significantly higher in multislice mode (95%) compared with multiplane mode (77%, P < .05). Diagnostic accuracy for detecting right CAD was also significantly higher in multislice mode (93% vs 80%, P < .05). CONCLUSION: Assessment of LV wall motion from multiple short-axis slices extracted from full volume RT3DE datasets improves the diagnosis of CAD and is thus a useful addition to DSE tools. PMID- 19307100 TI - Percutaneous closure of an atrial septal defect guided by live three-dimensional transesophageal echocardiography. AB - A 55-year-old man with a history of cryptogenic stroke presented to the authors' department for investigation. On transthoracic echocardiography, he was found to have a small secundum atrial septal defect, and transesophageal echocardiography was performed for a more detailed assessment. Following this, the defect was deemed suitable for percutaneous closure. The case demonstrates the utility and benefits of live three-dimensional transesophageal echocardiography for the assessment of this type of defect and guidance of transcatheter closure. PMID- 19307101 TI - Impact of energy loss coefficient on left ventricular mass regression in patients undergoing aortic valve replacement: preliminary observation. AB - BACKGROUND: The purpose of this study was to evaluate the impact of Doppler derived energy loss coefficient (ELCo) on the regression of left ventricular (LV) hypertrophy after aortic valve replacement (AVR) in patients with severe aortic stenosis. METHODS: Twenty-four patients with severe aortic stenosis who underwent AVR with Carpentier-Edwards pericardial bioprosthetic valves (valve size 19 mm, n = 16; valve size 21 mm, n = 8) were examined. Within 12 months after AVR, follow up echocardiography and Doppler measurements were performed. The effect of AVR was quantified on the basis of absolute and relative LV mass regression. RESULTS: There were significant correlations between indexed ELCo and absolute (r = 0.50, P = .013) and relative (r = 0.48, P = .018) LV mass regression. The mean value of relative LV mass regression was 25%, and a cutoff value of 0.9 cm2/m2 for indexed ELCo could detect patients with relative LV mass regression > 25% after AVR with sensitivity of 71% and specificity of 100%. CONCLUSIONS: ELCo, which can be calculated noninvasively from echocardiography, might be an important value to relate to LV mass regression in patients after AVR. PMID- 19307102 TI - Size-adjusted left ventricular outflow tract diameter reference values: a safeguard for the evaluation of the severity of aortic stenosis. AB - OBJECTIVE: We sought to evaluate the relationship among left ventricular outflow tract diameter (LVOTd), gender, and body surface area (BSA) and to evaluate the usefulness of size-adjusted LVOTd reference values in patients with aortic stenosis (AS). AS grading is based on the echocardiographic calculation of the aortic valve area (AVA) and requires LVOTd measurements, one main potential source of error. Transesophageal echocardiography (TEE) is reputed to be more accurate than transthoracic echocardiography (TTE), but validation studies are rare. A safeguard for LVOTd measurements is thus desirable. METHODS: Since January 2006, 3 subsets of patients have been prospectively and concurrently enrolled: 1) TEE group: In 120 patients with and without AS, we prospectively measured LVOTd during both TTE and TEE. 2) Validation set: In 382 patients without aortic valve or ascending aorta diseases, we evaluated the relationship among LVOTd, gender, and BSA. 3) Testing set: In 173 patients with AS, we compared the AVA obtained using measured LVOTd (AVA(MEAS)) and calculated LVOTd derived from a regression determined in the validation set (AVA(CALC)). RESULTS: TTE did not differ from and correlated well with TEE measurements overall (23 +/- 2 mm vs 23 +/- 2 mm, P = .26; r = 0.95, P < .0001) and in patients with AS (N = 43) (24 +/- 2 mm vs 24 +/- 3 mm, P = .15; r = 0.92, P < .0001). LVOTd was linearly correlated to BSA independently of gender (LVOTd = 5.7 * BSA+12.1; r = 0.55, P < .0001). In the testing set, AVA(CALC) did not differ from and correlated well with AVA(MEAS) (1.20 +/- 0.42 cm2 vs 1.23 +/- 0.40 cm2; P = .08; r = 0.89; P < .0001). CONCLUSION: TTE and TEE measurements of the LVOTd provided similar results. LVOTd was significantly associated to BSA and LVOTd, derived from a linear regression linked to BSA independently of gender, provided an acceptable approximation of the AVA. Thus, although accurate measurement of LVOTd is a crucial part of the echocardiographic evaluation of AS severity, the present equation may be used as a safeguard when this measurement is difficult or not possible with TTE. PMID- 19307104 TI - The weight of interleukin-6 in B cell-related autoimmune disorders. AB - Interleukin (IL)-6 is a prevailing factor of polyclonal B-cell activation of B cells, and thereby of their tolerance breach. Its receptor (R) complex consists of a transducing unit, and a membrane-bound or soluble protein. Many activities ascribed to this cytokine are generated by the soluble IL-6R. Evidence has however been gleaned in autoimmune diseases that the system is instrumental in rheumatoid arthritis (RA), Sjogren's syndrome and systemic lupus erythematosus (SLE). To gain insight into the understanding of the mechanisms behind these observations, a prime example is the recombination-activating gene (Rag) machinery in B lymphocytes. It is interesting that the expression of Rags is favored by IL-6, and repressed by anti-IL-6R antibody (Ab) in RA and SLE. Not surprisingly, clinical benefits are reported in the treatment of autoimmune disorders with anti-IL-6R Ab, and other perspectives about to be open in biotherapy. PMID- 19307105 TI - Inhibition of VEGF expression in tongue squamous cancer cells via RNA interference silencing of iNOS gene. AB - The purpose of this study was to investigate the regulatory role of the inducible nitric oxide synthase (iNOS) gene on vascular endothelial growth factor (VEGF) expression in oral squamous cancer cells. The RNA interference (RNAi) technique was used to silence iNOS gene expression by transfecting an expression vector containing short hairpin RNA (shRNA) for iNOS into Tca8113 tongue squamous cancer cells using cationic liposomes. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting were used to monitor iNOS and VEGF mRNA, as well as protein expression. iNOS mRNA expression was significantly downregulated 24 and 36 h after transfection, and iNOS protein expression was significantly downregulated at 36 and 48 h (P<0.05 versus control), showing that effective silencing was achieved. VEGF mRNA was significantly decreased 24 and 36 h post transfection, and VEGF protein expression was significantly decreased at 36 and 48 h (P<0.05). RNAi can decrease iNOS gene expression and achieve a gene silencing effect. iNOS gene silencing reduces VEGF expression levels in Tca8113 cells. Thus, there is a relationship between iNOS and VEGF expression in tongue squamous cancer cells. PMID- 19307103 TI - The etiology of autoimmune thyroid disease: a story of genes and environment. AB - Autoimmune thyroid diseases (AITDs), including Graves' disease (GD) and Hashimoto's thyroiditis (HT) are prevalent autoimmune diseases, affecting up to 5% of the general population. Autoimmune thyroid diseases arise due to complex interactions between environmental and genetic factors. Significant progress has been made in our understanding of the genetic and environmental triggers contributing to AITD. However, the interactions between genes and environment are yet to be defined. Among the major AITD susceptibility genes that have been identified and characterized is the HLA-DR gene locus, as well as non-MHC genes including the CTLA-4, CD40, PTPN22, thyroglobulin, and TSH receptor genes. The major environmental triggers of AITD include iodine, medications, infection, smoking, and possibly stress. Recent data on the genetic predisposition to AITD lead to novel putative mechanisms by which the genetic-environmental interactions may lead to the development of thyroid autoimmunity. PMID- 19307106 TI - Pharmacokinetics of azithromycin in plasma and sinus mucosal tissue following administration of extended-release or immediate-release formulations in adult patients with chronic rhinosinusitis. AB - This study compared the pharmacokinetics of azithromycin (AZI) following administration of extended-release (ER) and immediate-release (IR) formulations in plasma and sinus mucosa in patients with chronic rhinosinusitis. Patients (n=71) were randomised 1:1 to receive a single dose of AZI-ER 2g or up to three doses of AZI-IR 500 mg daily. Paired plasma and sinus tissue samples were taken during endoscopic sinus surgery at 2-168 h (four patients per time point) after the first dose. Samples were measured by a validated liquid chromatography/mass spectrometry assay. Pharmacokinetics were determined using composite concentration-time profiles. Comparison between formulations showed that within the first 24 h, the AZI area under the plasma concentration-time curve (AUC(24)) for ER was 5.2- and 7.0-fold higher than IR in plasma and sinus tissue, respectively. Comparison between matrices showed that the AUC(24) and AUC(168) in sinus tissue were 28.2- and 62.2-fold higher than in plasma for the ER formulation, whilst the AUC(24) in sinus tissue was 21.1-fold higher than in plasma for IR formulation. These results indicated that AZI has good penetration into sinus tissue regardless of formulation; however, dosing of AZI-ER (2 g) increased AZI exposure within the first 24 h compared with the Day 1 dose of 500 mg IR regimen. PMID- 19307107 TI - Pharmacokinetics and pharmacodynamics of biapenem in human cerebrospinal fluid. PMID- 19307108 TI - 25Mg NMR site analysis in metals and intermetallics. AB - Nuclear magnetic resonance (NMR) of the low abundance and low gyromagnetic ratio isotope (25)Mg, I=5/2, 2.606MHz/T, 10% abundant, is shown here to provide an informative probe for phase identification, site symmetry and site multiplicity of the intermetallic compounds which occur as strengthening precipitate phases in lightweight alloys. The intermetallics discussed here, Mg(17)Al(12), MgZn(2), Mg(2)Al(3) and Al(2)CuMg, are the final equilibrium precipitate phases in a number of Mg- and Al-based heat-treatable alloys. The (25)Mg spectra of Mg in Al 10at%Mg alloy show the progressive precipitation of Mg(2)Al(3) from Mg in solid solution as a function of annealing time at 150 degrees C. Also reported are (25)Mg spectra for CuMg(2), Mg(44)Al(15)Zn(41) and Mg(2)Sn, along with the counter atom (67)Zn and (63)Cu NMR spectra for MgZn(2) and CuMg(2). All spectra are simulated to determine nuclear interaction parameters and confirm site occupancy. PMID- 19307109 TI - Solid-state NMR and EPR analysis of carbon-doped titanium dioxide photocatalysts (TiO(2-)(x)C(x)). AB - Carbon-doped TiO(2) have received attention recently because of their potential for environmental photocatalysis and solar hydrogen conversion applications. Three different carbon-doped TiO(2) nanoparticle materials were synthesized via sol-gel and hydrothermal procedures, and analyzed by (13)C solid-state nuclear magnetic resonance (SSNMR) and other methods to characterize the environment of the doping species. UV/vis spectra and powder X-ray diffraction (XRD) patterns showed that the synthesized materials absorbed visible light and their crystal structures corresponded to anatase. (13)C SSNMR analyses of TiO(2-)(x)C(x) displayed signals corresponding to carbonate-type or sp(2)-type carbon species. Variable contact CP-MAS and dipolar dephasing analyses gave evidence for the presence and proximity of H atoms near these carbonate species. Electron paramagnetic resonance (EPR) spectroscopy showed that the thermally oxidized TiO(2-)(x)C(x) displayed a complex mixture of point defects, electron and hole trapping centers, all attributable to the incorporation of carbon, while the XPS data ruled out the presence of carbide species. PMID- 19307110 TI - Plasma fatty acid profiles in autism: a case-control study. AB - Increasing evidence is mounting in support of fatty acid metabolism playing a role in neurodevelopmental disorders such as autism. In order to definitely determine whether fatty acid concentrations were associated with autism, we quantitatively measured 30 fatty acids from seven lipid classes in plasma from a large subset of subjects enrolled in the Childhood Autism Risk from Genetics and the Environment (CHARGE) study. The CHARGE study is a large, population-based case-control study on children aged 2-5 born in California. Our subset consisted of 153 children with autism and 97 developmentally normal controls. Results showed that docosahexaenoic acid (DHA, 22:6n-3) was significantly decreased in phosphatidylethanolamine. Dimethyl acetals were significantly decreased in phosphatidylethanolamine and phosphatidylcholine as well. These results are consistent with the only other study to measure dimethyl acetals in children with autism, and suggest that the function of peroxisomes and the enzymes of the peroxisome involved with fatty acid metabolism may be affected in autism. PMID- 19307111 TI - Decrypting the genome's alternative messages. AB - Alternative splicing of messenger RNA (mRNA) precursors affects the majority of human genes, has a considerable impact on eukaryotic gene function and offers distinct opportunities for regulation. Alterations in alternative splicing can cause or modify the progression of a significant number of pathologies. Recent high-throughput technologies have uncovered a wealth of transcript diversity generated by alternative splicing, as well as examples for how this diversity can be established and become misregulated. A variety of mechanisms modulate splice site choice coordinately with other cellular processes, from transcription and mRNA editing or decay to miRNA-based regulation and telomerase function. Alternative splicing studies can contribute to our understanding of multiple biological processes, including genetic diversity, speciation, cell/stem cell differentiation, nervous system function, neuromuscular disorders and tumour progression. PMID- 19307112 TI - Enzyme-enhanced fluorescence detection of DNA on etched optical fibers. AB - A novel DNA biosensor based on enzyme-enhanced fluorescence detection on etched optical fibers was developed. The hybridization complex of DNA probe and biotinylated target was formed on the etched optical fiber, and was then bound with streptavidin labeled horseradish peroxidase (streptavidin-HRP). The target DNA was quantified through the fluorescent detection of bi-p,p'-4 hydroxyphenylacetic acid (DBDA) generated from the substrate 4 hydroxyphenylacetic acid (p-HPA) under the catalysis of HRP, with a detection limit of 1 pM and a linear range from 1.69 pM to 169 pM. It is facile to regenerate this sensor through surface treatment with concentrated urea solution. It was discovered that the sensor can retain 70% of its original activity after three detection-regeneration cycles. PMID- 19307113 TI - Energy use and recovery strategies within wastewater treatment and sludge handling at pulp and paper mills. AB - This paper presents an inclusive approach with focus on energy use and recovery in wastewater management, including wastewater treatment (WWT) and sludge handling. Process data from three Swedish mills and a mathematical model were used to evaluate seven sludge handling strategies. The results indicate that excess energy use in WWT processes counters the potential energy recovery in the sludge handling systems. Energy use in WWT processes is recommended to aim for sufficient effluent treatment, not for sludge reduction. Increased secondary sludge production is favourable from an energy point of view provided it is used as a substrate for heat, biogas or electricity production. PMID- 19307114 TI - A new class of 5-HT2B antagonists possesses favorable potency, selectivity, and rat pharmacokinetic properties. AB - We have been exploring the potential of 5-HT(2B) antagonists as a therapy for chronic heart failure. To assess the potential of this therapeutic approach, we sought compounds possessing the following attributes: (a) potent and selective antagonism of the 5-HT(2B) receptor, (b) low impact of serum proteins on potency, and (c) desirable pharmacokinetic properties. This Letter describes our investigation of a biphenyl benzimidazole class of compounds that resulted in 5 HT(2B) antagonists possessing the above attributes. Improving potency in a human serum albumin shift assay proved to be the most significant SAR discovery. PMID- 19307115 TI - Discovery of potent and selective DP1 receptor antagonists in the azaindole series. AB - Azaindole based structures were evaluated as DP1 receptor antagonists. This work has lead to the discovery of potent, selective and distinct DP1 receptor antagonists. PMID- 19307116 TI - GPR109a agonists. Part 1: 5-Alkyl and 5-aryl-pyrazole-tetrazoles as agonists of the human orphan G-protein coupled receptor GPR109a. AB - 5-Alkyl and aryl-pyrazole-tetrazoles have been identified as a new class of selective, small-molecule, agonists of the human G-protein-coupled receptor GPR109a, a high affinity receptor for the HDL-raising drug nicotinic acid. PMID- 19307117 TI - Facile synthesis of de-O-sulfated salacinols: revision of the structure of neosalacinol, a potent alpha-glucosidase inhibitor. AB - Facile synthesis of de-O-sulfated salacinols (3) was developed by employing the coupling reaction of an epoxide, 1,2-anhydro-3,4-di-O-benzyl-D-erythritol (9) with 2,3,5-tri-O-benzyl-1,4-dideoxy-1,4-epithio-D-arabinitol (10) as the key reaction. The reported structure of a potent alpha-glucosidase inhibitor named neosalacinol (8), isolated recently from Ayurvedic medicine Salacia oblonga, was proved incorrect, and revised to be de-O-sulfated salacinol formate (3c) by comparison of the spectroscopic properties with those of the authentic specimen synthesized. Discrepancies and confusion in the literature concerning the NMR spectroscopic properties of salacinol (1) have also been clarified. PMID- 19307118 TI - Synthesis of boronic acid derivatives of tyropeptin: proteasome inhibitors. AB - Boronic acid derivatives of tyropeptin were synthesized with TP-110 as the lead compound. Due to the lability of the aminoboronic acid moiety, careful design of the deprotection and coupling sequence was required. Liquid-liquid partition chromatography was found to be a powerful tool for purification of compounds of this class. The obtained derivatives showed potent inhibitory activities against the human 20S proteasome in vitro. PMID- 19307119 TI - Relationships between the structures of flavanone derivatives and their effects in enhancing early growth response-1 gene expression. AB - To identify the structural requirements that are pivotal in enhancing Early growth response-1 (Egr-1) expression, the quantitative relationships between the structural properties of flavanone derivatives and their increments of Egr-1 expression were elucidated using comparative molecular field analysis and comparative molecular similarity indices analysis. PMID- 19307120 TI - Quality of practice in an intensive care unit (ICU): a mini-ethnographic case study. AB - BACKGROUND: Nurses' perspectives of maintaining quality practice is important in the context of today's safety and quality agenda. This study provides a snapshot of registered nurses' perspectives on practice quality in one Australian ICU. METHODS: A mini-ethnographic case study examined how quality is embedded in the culture of ICU nursing. Semi-structured interviews of 10 informants were analysed using Spradley's (Spradley JP. The ethnographic interview. Sydney: Holt, Rinehart and Winston, Inc.;1979) ethnographic guidelines. FINDINGS: Three major themes influenced nursing quality: maintaining cohesiveness in a complex and stressful environment; rapid, effective and respectful communication; and specialist knowledge gained through experience and formal learning. CONCLUSION: The nurses reported satisfaction with the quality of their ICU practice, but revealed factors that could diminish quality and contribute to job dissatisfaction. Given current recruitment and retention issues and the link between satisfaction and retention, it is important to consider this these factors in advancing the knowledge base for workforce planning. PMID- 19307121 TI - Femoroacetabular impingement alters hip and pelvic biomechanics during gait Walking biomechanics of FAI. AB - Femoroacetabular impingement (FAI) has been reported to cause hip pain in a variety of daily activities including walking. However, the biomechanics of level gait has not been compared between FAI patients and a control group. This study quantified the affect of cam FAI on the three-dimensional (3-D) kinematics of the hip and pelvis, as well as the 3-D kinetics generated at the hip during walking. A unilateral cam impingement group (n=17) was compared to a matched control group (n=14) using between-group one-way ANOVAs. The FAI group had significantly lower peak hip abduction (p=0.009), frontal range of motion (ROM) (p=0.003), as well as attenuated pelvic frontal ROM (pelvic roll) (p=0.004) compared to the controls during level gait. There was also a trend of the impinged group having a lower sagittal ROM (p=0.047) than the controls. However, there were no kinetic differences between the two groups. Attenuated hip abduction, frontal ROM and sagittal ROM during gait in FAI individuals may be caused by soft tissue restriction, and decreased frontal pelvic ROM could result from limited mobility at the sacro-lumbar joint. PMID- 19307122 TI - Biased codon usage in signal peptides: a role in protein export. AB - The signal peptide of proteins exported via the general secretory pathway encodes structural features that enable the targeting and export of the protein to the periplasm. Recent studies have shown biased codon usage at the second amino acid position and a high usage of non-optimal codons within the signal peptide. Altering these biases in codon usage can have deleterious effects on protein folding and export. We propose that these codon-usage biases act in concert to optimize the export process through modulating ribosome spacing on the transcript. This highlights a new aspect of protein export and implies that codon usage in the signal peptide encodes signals that are important for protein targeting and export to the periplasm. PMID- 19307123 TI - Comparing anti-HIV, antibacterial, antifungal, micellar, and cytotoxic properties of tricarboxylato dendritic amphiphiles. AB - Three series of homologous dendritic amphiphiles--RCONHC(CH(2)CH(2)COOH)(3), 1(n); ROCONHC(CH(2)CH(2)COOH)(3), 2(n); RNHCONHC(CH(2)CH(2)COOH)(3), 3(n), where R = n-C(n)H(2n+1) and n = 13-22 carbon atoms--were assayed for their potential to serve as antimicrobial components in a topical vaginal formulation. Comparing epithelial cytotoxicities to the ability of these homologues to inhibit HIV, Neisseria gonorrhoeae, and Candida albicans provided a measure of their prophylactic/therapeutic potential. Measurements of the ability to inhibit Lactobacillus plantarum, a beneficial bacterium in the vagina, and critical micelle concentrations (CMCs), an indicator of the potential detergency of these amphiphiles, provided additional assessments of safety. Several amphiphiles from each homologous series had modest anti-HIV activity (EC(50) = 110-130 microM). Amphiphile 2(18) had the best anti-Neisseria activity (MIC =65 microM), while 1(19) and 1(21) had MICs against C. albicans of 16 and 7.7 microM, respectively. Two measures of safety showed promise as all compounds had relatively low cytotoxic activity (EC(50) = 210-940 microM) against epithelial cells and low activity against L. plantarum, 1(n), 2(n), and 3(n) had MICs490, 1300, and 940 microM, respectively. CMCs measured in aqueous triethanolamine and in aqueous potassium hydroxide showed linear dependences on chain length. As expected, the longest chain in each series had the lowest CMC-in triethanolamine: 1(21), 1500 microM; 2(22), 320 microM; 3(22), 340 microM, and in potassium hydroxide: 1(21), 130 microM; 3(22), 40 microM. The CMC in triethanolamine adjusted to pH 7.4 was 400 microM for 1(21) and 3900 microM for 3(16). The promising antifungal activity, low activity against L. plantarum, relatively high CMCs, and modest epithelial cytotoxicity in addition to their anti-Neisseria properties warrant further design studies with dendritic amphiphiles to improve their safety indices to produce suitable candidates for antimicrobial vaginal products. PMID- 19307124 TI - Synthesis and anti-hepatitis B virus activities of Matijing-Su derivatives. AB - A series of derivatives of Matijing-Su (MTS, N-(N-benzoyl-L-phenylalanyl)-O acetyl-L-phenylalanol) was synthesized and evaluated for their anti-hepatitis B virus (HBV) activities in 2.2.15 cells. The IC(50) of compounds 9c (1.40 microM), 9g (2.33 microM) and 9n (2.36 microM), etc. and the selective index of 9n (45.93) of the inhibition on the replication of HBV DNA were higher than those of the positive control lamivudine [41.59, (IC(50): 82.42 microM)]. Compounds 11d, 12a and 12e also exhibited significant anti-HBV activities. PMID- 19307125 TI - Excitation function of the 192Os(3He,4n)-reaction for production of 191Pt. AB - In search of an alternative production route of the therapeutically and environmentally interesting radionuclide (191)Pt (T(1/2)=2.8 d), excitation function of the (192)Os((3)He,4n)(191)Pt reaction was measured from its threshold up to 36 MeV. Thin samples of enriched (192)Os were prepared by electrodeposition on Ni-foils, and the conventional stacked-foil technique was used for cross section measurements. The experimental data were compared with the results of theoretical calculations using the codes ALICE-IPPE and TALYS. Good agreement was found with TALYS. The theoretical thick target yield of (191)Pt over the energy range E(3He)=36-->25 MeV amounts to 6.7 MBq/microA h. A comparison of various investigated production methods of (191)Pt is given. The here investigated (192)Os((3)He,4n)-process leads to very high-purity (191)Pt (>99.5%). PMID- 19307126 TI - Synthesis of Pt-Ru@PThB catalyst by gamma-irradiation and NaBH(4) as reducing agent. AB - Pt-Ru particles were loaded onto spherical polythiophene balls (PThB) and new catalysts, Pt-Ru@PThBs, were prepared. Pt and Ru ions were subjected to chemical reduction using NaBH(4) as reducing agent or gamma-irradiation in the presence of PThBs. The size, morphology, and composition of Pt-Ru@PThB catalysts were determined by transmission electron microscope (TEM), X-ray diffraction (XRD) and elemental analysis. TEM images of the Pt-Ru@PThB catalysts clearly provide evidences that Pt-Ru alloy nanoparticles are dispersed onto the surface of PThBs. The electrocatalytic efficiency of Pt-Ru@PThB was tested for electrochemical oxidation of carbon monoxide as well as for methanol. Pt-Ru@PThB catalyst prepared by gamma-irradiation exhibits the enhanced electrocatalytic oxidation of CO and methanol. We envisage that Pt-Ru@PThB catalysts are suited for direct methanol fuel cells. PMID- 19307127 TI - Tritium production, recovery and application in Korea. AB - Four CANDU reactors have been operating at the site of Wolsong Nuclear Power Generation in Korea. The Wolsong tritium removal facility was constructed to reduce the tritium levels in heavy water systems. This facility was designed to process 100kg/h of tritiated heavy water feed and to produce 99% pure T(2). This recovered tritium will be made available for commercial applications. The initial phases on the tritium applications are made to establish the infrastructure and the tritium controls. PMID- 19307128 TI - A comparison of dose distributions of HDR intracavitary brachytherapy using different sources and treatment planning systems. AB - To evaluate the influence of different sources and treatment planning systems (TPSs), we calculated the dose distributions of 10 patients treated by HDR intracavitary brachytherapy. The dose distributions of treatment plans were compared to the points A, B and ICRU bladder and rectum reference points located in the relevant clinical planes. The results show that the dose discrepancies between the two treatment plans are mainly affected by the differences in the physical characteristics of source and the positioning method in TPSs. PMID- 19307129 TI - Alternative method for (64)Cu radioisotope production. AB - The method for (64)Cu production based on a (64)Ni target using an 18MeV proton energy beam was developed. The studies on the optimisation of targetry for the 18MeV proton bombardments were performed in terms of the cost-effective target utilisation and purity of the (64)Cu product. The thickness-specific (64)Cu yield (microCi/(microA x microm)) was introduced into the optimisation calculation with respect to cost-effective target utilisation. A maximum target utilisation efficacy factor (TUE) was found for the proton energy range of 2.5-13MeV with corresponding target thickness of 36.2microm. With the optimised target thickness and proton energy range, the (64)Ni target thickness saving of 45.6% was achieved, while the overall (64)Cu yield loss is only 23.9%, compared to the use of the whole effective proton energy range of 0-18MeV with target thickness of 66.6microm. This optimisation has the advantage of reducing the target amount to a reasonable level, and therefore the cost of the expensive (64)Ni target material. The (64)Ni target electroplated on the Au-Tl multi layer coated Cu substrate was a new and competent design for an economic production of high quality (64)Cu radioisotope using an 18MeV proton energy cyclotron or a 30MeV cyclotron with proton beam adjustable to 18MeV. In this design, the Au coating layer plays a role of protection of "cold" Cu leakage from the Cu substrate and Tl serves to depress the proton beam energy (from 18MeV to the energy optimised value 13MeV). The ion exchange chromatographic technique with a gradient elution was applied to improve the (64)Cu separation with respect to reducing the processing time and control of (64)Cu product quality. PMID- 19307130 TI - The robotic radiosynthesis of 5-[(18)F]fluoro-2'-deoxyuridine and its biological characterization. AB - 5-[(18)F]fluoro-2'-deoxyuridine ([(18)F]FUdR) was synthesized using a robotic system as a proliferation probe for PET. [(18)F]FUdR was prepared via radiofluorodestannylation reaction from its organotin precursor. Biodistribution study and microPET imaging of [(18)F]FUdR in NG4TL4 sarcoma-bearing FVB/n mice were performed. The tumor-to-blood and tumor-to-muscle ratio increased steadily from 15 (1.81 and 3.42) to 120min (9.10 and 11.9) post injection. The dynamic microPET imaging demonstrates remarkable radioactivity retention in the tumor, which is consistent with the results of biodistribution study. PMID- 19307131 TI - A step function model to evaluate the real monetary value of man-sievert with real GDP. AB - For use in a cost-benefit analysis to establish optimum levels of radiation protection in Korea under the ALARA principle, we introduce a discrete step function model to evaluate man-sievert monetary value in the real economic value. The model formula, which is unique and country-specific, is composed of real GDP, the nominal risk coefficient for cancer and hereditary effects, the aversion factor against radiation exposure, and average life expectancy. Unlike previous researches on alpha-value assessment, we show different alpha values in the real term, differentiated with respect to the range of individual doses, which would be more realistic and informative for application to the radiation protection practices. GDP deflators of economy can reflect the society's situations. Finally, we suggest that the Korean model can be generalized simply to other countries without normalizing any country-specific factors. PMID- 19307132 TI - The synthesis and characterization of [(124)I]IMPY, a thioflavin-S derivative, in transgenic mouse models of Alzheimer's disease. AB - 6-[(124)I]iodo-2-(4'-N,N-dimethylamino)-phenylimidazo[1,2-a]pyridine ([(124)I]IMPY) was synthesized and characterized as a positron-emitting probe to identify Alzheimer's disease in transgenic mouse models. A significant reduction in radioactivity retention in the hippocampus and frontal cortex by co-incubation with nonradioactive IMPY was observed. Highly specific retention of radioactivity in beta-amyloid-rich regions of brain sections was also noted. This study demonstrated that [(124)I]IMPY was a promising probe for the mouse model and may be useful for positron emission tomography to image beta-amyloid plaques in the human brain. PMID- 19307133 TI - Evidence of a sirtuin gene family in grapevine (Vitis vinifera L.). AB - Sirtuins are known as regulators of age-dependent gene transcription and chromatin modification in yeast and in animals, but information about their occurrence and role in plants is scarce. Sirtuin-like sequences were amplified using two highly degenerate primers designed comparing sirtuin sequences of seven different plant species, and characterised at the sequence level in Vitis vinifera L. cv. Barbera clone R4. The data were confirmed and analysed by searching the published database from two independent grape genome projects with the obtained sequences. Two putative sirtuin genes containing sirtuin-conserved domains were found on chromosome 7 and on chromosome 19, suggesting the occurrence of sirtuins in grapevine. PMID- 19307134 TI - Is EuroSCORE useful in the prediction of extended intensive care unit stay after cardiac surgery? AB - OBJECTIVE: Risk stratification allows preoperative assessment of cardiac surgical risk faced by individual patients and permits retrospective analysis of postoperative complications in the intensive care unit (ICU). The aim of this single-center study was to investigate the prediction of extended ICU stay after cardiac surgery using both the additive and logistic model of the European System for Cardiac Operative Risk Evaluation (EuroSCORE). METHODS: A retrospective observational study was conducted. We collected clinical data of 1562 consecutive patients undergoing cardiac surgery over a 2-year period at the Antwerp University Hospital, Belgium. EuroSCORE values of all patients were obtained. The outcome measure was the duration of ICU stay in days. The predictive performance of EuroSCORE was analyzed by the discriminatory power of a receiver operating characteristic (ROC) curve. Each EuroSCORE value was used as a theoretical cut off point to predict duration of ICU stay. Three subsequent ICU stays were defined as prolonged: more than 2, 5 and 7 days. ROC curves were constructed for both the additive and logistic model. RESULTS: Patients had a median ICU stay of 2 days and a mean ICU stay of 5.5 days. Median additive EuroSCORE was 5 (range, 0 22) and logistic EuroSCORE was 3.94% (range, 0.00-87.00). In the additive EuroSCORE model, a predictive value of 0.76 for an ICU stay of >7 days, 0.72 for >5 days and 0.67 for >2 days was found. The logistic EuroSCORE model yielded an area under the ROC curve of 0.77, 0.75 and 0.68 for each ICU length of stay, respectively. CONCLUSIONS: In our patient database, prolonged length of stay in the ICU correlated positively with EuroSCORE. The logistic model was more discriminatory than the additive in tracing extended ICU stay. The overall predictive performance of EuroSCORE is acceptable and most likely based on the presence of variables that are risk factors for both mortality and extended ICU stay. Hence, EuroSCORE is a useful predicting tool and provides both surgeons and intensivists with a good estimate of patient risk in terms of ICU stay. PMID- 19307135 TI - Images in cardio-thoracic surgery: Intimo-intimal intussusception: a rare complication of Stanford type A acute aortic dissection. PMID- 19307137 TI - Chronic post-thoracotomy pain: a critical review of pathogenic mechanisms and strategies for prevention. AB - Chronic pain complaints after thoracic surgery represent a significant clinical problem in 25-60% of patients. Results from thoracic and other surgical procedures suggest multiple pathogenic mechanisms that include pre-, intra-, and postoperative factors. This review attempts to analyse the methodology and systematics of the studies on the post-thoracotomy pain syndrome (PTPS) after lung cancer surgery in adults, in order to clarify the relative role of possible pathogenic factors and to define future strategies for prevention. Literature published from 2000 to 2008 together with studies included in previous systematic reviews was searched recursively using PubMed and OVID by combining three categories of search terms. The available data have major inconsistencies in collection of pre-, intra- and postoperative data that may influence PTPS, thereby hindering precise conclusions as well as preventive and treatment strategies. However, intercostal nerve injury seems to be the most important pathogenic factor. Since there is a general agreement on the clinical relevance of PTPS, a proposal for design of future trials is presented. PMID- 19307138 TI - Adjuvant bisphosphonate therapy for breast cancer patients: Standard of care or future direction? AB - Bone is the most common site of metastatic spread in breast cancer patients. The use of bisphosphonates (BPs) in women with bone metastases from breast cancer has been shown to reduce the incidence, and delay the onset of, skeletal-related events. Indeed, BPs are now an established standard of care in treating patients with bone metastases. As with many new therapies, once efficacy in the metastatic setting is demonstrated, therapies are tested in the adjuvant setting. There are a number of trials that have tested the hypothesis that BP therapy in women with early breast cancer may not only reduce the development of skeletal metastases but also improve both disease-free and overall survival; recent randomized trials have demonstrated intriguing results regarding the possible anticancer effects of adjuvant BP therapy. Furthermore, interesting signals are filtering through from trials evaluating the role of BPs to prevent cancer-treatment induced bone loss. Many of these trials will be reported in the near future. In this article we review possible subgroups that may benefit from adjuvant treatment with BPs, and ongoing trials that may offer more definitive answers. PMID- 19307139 TI - Are dentists risking losing their relevance? PMID- 19307140 TI - Spin echo SPI methods for quantitative analysis of fluids in porous media. AB - Fluid density imaging is highly desirable in a wide variety of porous media measurements. The SPRITE class of MRI methods has proven to be robust and general in their ability to generate density images in porous media, however the short encoding times required, with correspondingly high magnetic field gradient strengths and filter widths, and low flip angle RF pulses, yield sub-optimal S/N images, especially at low static field strength. This paper explores two implementations of pure phase encode spin echo 1D imaging, with application to a proposed new petroleum reservoir core analysis measurement. In the first implementation of the pulse sequence, we modify the spin echo single point imaging (SE-SPI) technique to acquire the k-space origin data point, with a near zero evolution time, from the free induction decay (FID) following a 90 degrees excitation pulse. Subsequent k-space data points are acquired by separately phase encoding individual echoes in a multi-echo acquisition. T(2) attenuation of the echo train yields an image convolution which causes blurring. The T(2) blur effect is moderate for porous media with T(2) lifetime distributions longer than 5 ms. As a robust, high S/N, and fast 1D imaging method, this method will be highly complementary to SPRITE techniques for the quantitative analysis of fluid content in porous media. In the second implementation of the SE-SPI pulse sequence, modification of the basic measurement permits fast determination of spatially resolved T(2) distributions in porous media through separately phase encoding each echo in a multi-echo CPMG pulse train. An individual T(2) weighted image may be acquired from each echo. The echo time (TE) of each T(2) weighted image may be reduced to 500 micros or less. These profiles can be fit to extract a T(2) distribution from each pixel employing a variety of standard inverse Laplace transform methods. Fluid content 1D images are produced as an essential by product of determining the spatially resolved T(2) distribution. These 1D images do not suffer from a T(2) related blurring. The above SE-SPI measurements are combined to generate 1D images of the local saturation and T(2) distribution as a function of saturation, upon centrifugation of petroleum reservoir core samples. The logarithm mean T(2) is observed to shift linearly with water saturation. This new reservoir core analysis measurement may provide a valuable calibration of the Coates equation for irreducible water saturation, which has been widely implemented in NMR well logging measurements. PMID- 19307141 TI - H-start for exclusively heteronuclear NMR spectroscopy: the case of intrinsically disordered proteins. AB - Here, we present a series of exclusively heteronuclear multidimensional NMR experiments, based on 13C direct detection, which exploit the (1)H polarization as a starting source to increase the signal-to-noise ratio. This contributes to make this spectroscopy more useful and usable. Examples are reported for a suitable system such as securin, an intrinsically disordered protein of 22 kDa. PMID- 19307142 TI - Lentiviral-mediated correction of MPS VI cells and gene transfer to joint tissues. AB - Joint disease in mucopolysaccharidosis type VI (MPS VI) remains difficult to treat despite the success of enzyme replacement therapy in treating other symptoms. In this study, the efficacy of a lentiviral vector to transduce joint tissues and express N-acetylgalactosamine-4-sulphatase (4S), the enzyme deficient in MPS VI, was evaluated in vitro and the expression of beta-galactosidase was used to evaluate transduction in vivo. High viral copy number was achieved in MPS VI fibroblasts and 4-sulphatase activity reached 12 times the normal level. Storage of accumulated glycosaminoglycan was reduced in a dose dependent manner in both MPS VI skin fibroblasts and chondrocytes. Enzyme expression was maintained in skin fibroblasts for up to 41 days. Comparison of two promoters; the murine phosphoglycerate kinase gene promoter (pgk) and the myeloproliferative sarcoma virus long terminal repeat promoter (mpsv), demonstrated a higher level of marker gene expression driven by the mpsv promoter in both chondrocytes and synoviocytes in vitro. When injected into the rat knee, the expression of beta galactosidase from the mpsv promoter was widespread across the synovial membrane and the fascia covering the cruciate ligaments and meniscus. No transduction of chondrocytes or ligament cells was observed. Transduction was maintained for at least 8 weeks after injection. These results indicate that the lentiviral vector can be used to deliver 4S to a range of joint tissues in vitro and efficiently transduce synovial cells and express beta-galactosidase in vivo. PMID- 19307143 TI - Cholesteryl Ester Storage Disease (CESD) due to novel mutations in the LIPA gene. AB - Cholesteryl Ester Storage Disease (CESD) is a rare recessive disorder due to mutations in LIPA gene encoding the lysosomal acidic lipase (LAL). CESD patients have liver disease associated with mixed hyperlipidemia and low plasma levels of high-density lipoproteins (HDL). The aim of this study was the molecular characterization of three patients with CESD. LAL activity was measured in blood leukocytes. In two patients (twin sisters) the clinical diagnosis of CESD was made at 9 years of age, following the fortuitous discovery of elevated serum liver enzymes in apparently healthy children. They had mixed hyperlipidemia, hepatosplenomegaly, reduced LAL activity (approximately 5% of control) and heteroalleic mutations in LIPA gene coding sequence: (i) the common c.894 G>A mutation and (ii) a novel nonsense mutation c.652 C>T (p.R218X). The other patient was an 80 year-old female who for several years had been treated with simvastatin because of severe hyperlipidemia associated with low plasma HDL. In this patient the sequence of major candidate genes for monogenic hypercholesterolemia and hypoalphalipoproteinemia was negative. She was found to be a compound heterozygote for two LIPA gene mutations resulting in 5% LAL activity: (i) c.894 G>A and (ii) a novel complex insertion/deletion leading to a premature termination codon at position 82. These findings suggest that, in view of the variable severity of its phenotypic expression, CESD may sometimes be difficult to diagnose, but it should be considered in patients with severe type IIb hyperlipidemia associated with low HDL, mildly elevated serum liver enzymes and hepatomegaly. PMID- 19307144 TI - Transient leukoerythroblastosis in a very low birth weight infant with parvovirus B19 infection. AB - BACKGROUND: Leukoerythroblastosis is characterized by the presence of leukocytosis and erythroid and myeloid blast cells in the peripheral blood. The most common etiological factors of leukoerythroblastosis occurring during early childhood are viral infections, juvenile myelomonocytic leukemia, and osteopetrosis. To our knowledge, an association with parvovirus B19 infection has only been reported in a preterm infant. Human parvovirus B19 has been associated with red cell aplasia, leukopenia, and thrombocytopenia. CASE REPORT: The case of a very low birth weight preterm infant with transient leukoerythroblastosis associated with parvovirus B19 infection is described. CONCLUSIONS: Leukoerythroblastosis has to be kept in mind if a very high leukocyte count is detected in the neonatal period, and parvovirus B19 infection should be taken into consideration as the etiological factor for this entity. PMID- 19307145 TI - Innovation in a backwater: The Harpurhey Resettlement Team and the mental health services of North Manchester, 1982-1987. AB - This paper explores the circumstances around the setting up of the Harpurhey Resettlement Team, an innovative project which, in the late 1980s, resettled around 20 long-stay patients from Springfield Hospital in North Manchester into ordinary tenancies within the same neighbourhood. It argues that Springfield's position as a marginalised and neglected institution produced the conditions for such innovation; while the particular and unexpected convergence of national policies, local structures and institutional politics created space for a process of change which, in both form and outcome, could not have occurred in the more regulated psychiatric environments elsewhere in Manchester. PMID- 19307146 TI - Economic, racial and ethnic disparities in breast cancer in the US: towards a more comprehensive model. AB - Using cancer registry data, we focus on racial and ethnic disparities in stage of breast cancer diagnosis in Cook County, IL. The county health system is the "last resort" health-care provider for low-income persons. Socioeconomic status is measured using empirical Bayes estimates of tract-level poverty, specific to non Hispanic whites, non-Hispanic blacks or Hispanics in one of three age groups. We use ordinal logistic regression with non-proportional odds to model stage. Blacks and Hispanics are at greater risk for regional and distant stage diagnosis, but the disparity declines with age. Women in high-poverty areas are at substantially greater risk for late-stage diagnosis. The effects of poverty do not differ by age or across racial and ethnic groups. PMID- 19307147 TI - New tricks from ancient algae: natural products biosynthesis in marine cyanobacteria. AB - Cyanobacteria, among Earth's oldest organisms, have evolved sophisticated biosynthetic pathways to produce a rich arsenal of bioactive natural products. In consequence, cyanobacterial secondary metabolites have been an incredibly fruitful source of lead compounds in drug discovery efforts. Investigations into the biochemistry responsible for the creation of these compounds, complemented by genome sequencing efforts, are revealing unique enzymatic mechanisms not described or rarely described elsewhere in the natural world. Herein, we discuss recent advances in understanding the biosynthesis of three cyanobacterial classes of natural product: mixed polyketide synthase/non ribosomal peptide synthetase (PKS/NRPS) metabolites, aromatic amino acid-derived alkaloids, and ribosomally encoded cyclic peptides. The unique biosynthetic mechanisms employed by cyanobacteria are inspiring new developments in heterologous gene expression and biotechnology. PMID- 19307148 TI - The alignment of information systems with organizational objectives and strategies in health care. AB - PURPOSE: The alignment of information systems with organizational objectives and strategies is a key, contemporary challenge to organizations in general and the health care industry in particular. Researchers and managers alike believe that the selection of new information systems to support objectives and strategies focuses the organization on accomplishing its objectives and realizing the value of the investments in the systems. The purpose of this study was to help understand alignment in health care so that health care information systems planners can better achieve it. METHODS: Structured interviews with 15 top information systems managers in health care organizations of various sizes and types inquired about organizational objectives and strategies, the process for choosing new information systems to support those objectives and strategies, and the concomitant facilitating and hindering managerial actions and organizational characteristics. RESULTS: In addition to identifying and elucidating specific objectives, strategies, processes for choosing new systems, and facilitating and hindering actions and characteristics, the study used the data to characterize a generalized process of alignment in health care organizations. CONCLUSIONS: The study contributes by confirming that alignment is a significant issue in health care organizations, and that such organizations make deliberate efforts to achieve it. The study further contributes by providing tables of actions and characteristics that managers might use as checklists in current and future alignment efforts as well as in generally cultivating broad support for alignment. Finally, it contributes by suggesting future study of alignment's predictors and effects in health care organizations. PMID- 19307149 TI - Domain-specific analytical language modeling--the chief complaint as a case study. AB - PURPOSE: A large share of the information in electronic medical records (EMRs) consists of free-text compositions. From a computational point-of-view, the continuing prevalence of free-text entry is a major hindrance when the goal is to increase automation in EMRs. However, the efforts in developing standards for the structured representation of medical information have not proven to be a panacea. The information space of clinical medicine is very diverse and constantly evolving, making it challenging to develop standards for the domain. This paper reports a study aiming to increase automation in the EMR through the computational understanding of specific class of medical text in English, namely emergency department chief complaints. METHODS: We apply domain-specific analytical modeling for the computational understanding of chief complaints. We evaluate the performance of this approach in the automatic classification of chief complaints, e.g., for use in automatic syndromic surveillance. RESULTS: The evaluation in a multi-hospital setting showed that the presented algorithm was accurate in terms of classification correctness. Also, use of approximate matching in the algorithm to cope with typographic variance did not affect classification correctness while increasing classification completeness. PMID- 19307150 TI - Detection of mucosal and cutaneous human papillomaviruses in oesophagitis, squamous cell carcinoma and adenocarcinoma of the oesophagus. AB - BACKGROUND: Epidemiologic evidence points to a connection between viral infections by the human papillomavirus (HPV) and a subgroup of squamous cell carcinomas of the oropharynx. Still controversial is the association of HPV infection with oesophageal neoplasia. OBJECTIVES: To investigate the presence of mucosal as well as cutaneous HPVs in squamous cell carcinoma and adenocarcinoma of the oesophagus. STUDY DESIGN: HPV DNA has been searched by PCR and characterized by nucleotide sequence analysis in paraffin-embedded biopsies from Italian patients with oesophageal squamous cell carcinoma (n=36), sarcomatoid cell carcinoma (n=1), adenocarcinoma (n=20) and oesophagitis lesions (n=27). RESULTS: A broad spectrum of HPVs, primarily cutaneous types was demonstrated in 27.8% (10/36) of squamous cell carcinomas with a significantly higher frequency in well (G1) and moderately (G2) differentiated grades (47.3%, 9/19) compared to poorly (G3) differentiated (5.9%, 1/17) squamous cell carcinoma (p=0.008), and in 10% (2/20) of adenocarcinomas and in 29.6% (8/27) of oesophagitis. HPV types detected included mucosal types HPV 6 and 16, cutaneous types HPV 8, 15, 20 and 25; and the putative new HPV types X14, X15, DL473, PPHL1FR and CJ198. CONCLUSIONS: There is no evidence of any association between mucosal HPVs and oesophageal neoplasia. The cutaneous HPVs are detected at low frequency in adenocarcinoma and poorly differentiated squamous cell carcinoma, while they are frequently detected in oesophagitis and in well and moderately differentiated squamous cell carcinoma suggesting their tropism for keratinized tissue, although a significant association with such neoplasias cannot be drawn. PMID- 19307151 TI - Circadian rhythm disruption and reverse paradoxical insomnia after intraventricular hemorrhage. PMID- 19307152 TI - Three-year follow-up on the accumulation of cardiac (123)I-MIBG scintigraphy in idiopathic REM sleep behavior disorder. PMID- 19307153 TI - Patho-physiology of restless legs syndrome: a very tedious puzzle! PMID- 19307154 TI - The dopaminergic neurons of the A11 system in RLS autopsy brains appear normal. AB - Although the positive clinical benefits of levodopa have fostered the concept of an abnormality in the dopaminergic system in Restless Legs Syndrome (RLS), research into the nigro-striatal (PET/SPECT studies) or tubero-infundibular (i.e., prolactin secretion) dopaminergic pathways has shown limited positive results. Some research groups have focused on the A11 dopaminergic system in the hypothalamus as this is the primary source of descending dopaminergic input into the spinal cord, an area of the nervous system believed by some investigators to be involved in RLS symptom development. Some investigators have now proposed lesioning or toxin-inhibiting the A11 system as a model of RLS, even though there has been no clear clinical or autopsy data to suggest that RLS is a neurodegenerative disorder. In this study, the A11 cell bodies were identified in 6 RLS and 6 aged-matched control autopsy cases. Cells were stained for tyrosine hydroxylase (TH), and stereological measure of the individual TH (+) cell volume was made. Regional assessment of gliosis as assessed by immunostaining for glial fibrillary acidic protein (GFAP) was made in the surrounding tissue. General histological staining was also performed on the tissue. This study found no significant difference between RLS or control cases on any measure used: TH (+) cell volume, fractional GFAP staining, or general histological examination. Nor was there histological indication of any significant inflammation or concurrent ongoing pathology in these RLS cases. The findings do not support the concept of dramatic cell loss or of a neurodegenerative process in the A11 hypothalamic region of patients with RLS. However, that does not exclude the possibility that the A11 system is involved in RLS symptoms. Changes at the cellular level in dopaminergic metabolism or at the distal synapse with changes in receptors or transporters were not evaluated in this study. PMID- 19307155 TI - Prevalence and risk factors of obstructive sleep apnea among middle-aged urban Indians: a community-based study. AB - BACKGROUND: The epidemiology of obstructive sleep apnea (OSA) and obstructive sleep apnea syndrome (OSAS) is not well studied among Indians. We sought to determine the prevalence and risk factors of OSA in a middle-aged urban Indian population. METHODS: We conducted a two-stage, cross-sectional, community-based study in four different socioeconomic zones of the South Delhi district, India, from April 2005 to June 2007. In stage 1, subjects of either gender aged 30-65 yrs were administered a questionnaire by door-to-door survey using systematic random sampling. Subjects that responded were classified as habitual and non habitual snorers. In stage 2, all the habitual and 10% of randomly selected non habitual snorers were invited for overnight polysomnographic study. RESULTS: Of the 2860 subjects approached, 2505 (88%) completed stage 1. Habitual snoring was present in 452 (18%) subjects. In stage 2, OSA defined as apnea-hypopnea index 5 was observed in 94 (32.4%) of 290 habitual snorers and 3 (4%) of 75 non-habitual snorers. Estimated population prevalence of OSA and OSAS was 9.3% (95% CI 8.2 10.5%) and 2.8% (2.1-3.4%) respectively. On multivariable analysis, male gender (adjusted odds ratio 3.8 [1.7-4.9]), body-mass index 25 kg/m(2) (4.1 [2.0-8.3]), and abdominal obesity (2.2 [1.9-5.3]) were independently associated with the presence of OSA. A linear trend was observed in the prevalence of OSA across the socioeconomic strata. CONCLUSIONS: OSA is a significant public health problem in the middle-aged Indian population across the socioeconomic spectrum. OSA is associated with some of the well known risk factors for cardiovascular disease. PMID- 19307156 TI - T-bet expression is regulated by EGR1-mediated signaling in activated T cells. AB - T-bet is a Th1-specific transcription factor that is directly involved in three important pathways for Th1 cell differentiation, namely TCR signaling, and the IFN-gamma-STAT1 and IL-12-STAT4 pathways. A recent study also showed that T-bet plays a vital role in innate immunity. However, the molecular mechanism responsible for transcriptional activation of T-bet during T cell development is not yet known. Here, we characterize the essential human T-bet promoter elements and show that binding of EGR1 to this promoter induces T-bet transcription. Notably, overexpression of EGR1 transactivates and, synergistically in concert with TCR signaling, induces T-bet expression in activated T cells. In contrast, depletion of EGR1 significantly decreases T-bet induction. Finally, we report a positive correlation between EGR1 and T-bet expression during T helper cell differentiation. Collectively, these findings provide molecular insight into T bet transcription and suggest that EGR1 is an upstream regulator of T-bet induction. PMID- 19307157 TI - Increased chitotriosidase activity in serum of leprosy patients: association with bacillary leprosy. AB - Human phagocyte-specific chitotriosidase is associated with several diseases involving macrophage activation. Since macrophage activation plays an important role in the control of Mycobacterium leprae infection, we studied the association of chitotriosidase with leprosy both in serum and in situ in lesional skin biopsies from patients. Serum samples from 78 Indonesian leprosy patients (39 non reactional and 39 reactional leprosy patients) and 36 healthy controls (HC) from the same endemic region were investigated. The patients were classified as multibacillary (MB, n=69) or paucibacillary (PB, n=9) based on the bacterial index in slit-skin smears. Thirty-six of the reactional patients had erythema nodosum leprosum (ENL), while only 3 had reversal reaction (RR). Follow-up serum samples after corticosteroid treatment were also obtained from 17 patients with ENL and one with RR. Multibacillary (MB) patients showed increased chitotriosidase activity in serum as compared to paucibacillary (PB) patients and healthy controls. Although no significant difference was observed between reactional and the corresponding non-reactional groups, ENL showed significantly higher chitotriosidase activity as compared to HC. Furthermore, corticosteroid treatment resulted in significant decline of enzyme activity in ENL sera. Chitotriosidase activity correlated with levels of neopterin, another macrophage activation marker, but not with IL-6, IFN-gamma, TNF-alpha and IL-10. Immunohistochemical staining of 6 MB (LL=5, BL=1) lesional skin sections from stored material showed positive staining for chitotriosidase within lipid-laden macrophages suggesting that macrophages are the source of the enzyme detected in serum. Thus, serum chitotriosidase activity is potentially useful in distinguishing MB from PB leprosy and in monitoring response to therapy in ENL. PMID- 19307158 TI - Nitric oxide synthase isozymes in spinally projecting PVN neurons are involved in CRF-induced sympathetic activation. AB - In the brain, corticotropin-releasing factor (CRF) has been shown to activate the sympatho-adrenomedullary outflow, but the central mechanisms of action are still not fully understood. Previously, we reported that inducible nitric oxide synthase (iNOS) is involved in central CRF-induced elevation of plasma catecholamines in rats. Nitric oxide is mainly synthesized by neuronal NOS (nNOS) and iNOS in many areas in the brain. Of these areas, the paraventricular hypothalamic nucleus (PVN) contains neurons projecting to the intermediolateral cell column (IML) of the spinal cord, thereby directly affecting the sympathetic activity. Therefore, in the present study, we investigated the effect of intracerebroventricularly (i.c.v.) administered CRF on plasma catecholamine levels and expression of NOS isozymes (iNOS and nNOS) and Fos (a marker for neuronal activation) in the spinally projecting PVN neurons, using rats microinjected with a monosynaptic retrograde tracer into the IML. CRF (1.5 nmol/animal, i.c.v.) effectively elevated plasma catecholamine levels. The spinally projecting neurons labeled with a tracer were detected in the dorsal cap, ventral part and posterior part of the PVN. CRF significantly increased the number of spinally projecting neurons triple-labeled with Fos and iNOS in all of these PVN subnuclei. On the other hand, CRF significantly increased the number of spinally projecting neurons triple-labeled with Fos and nNOS only in the ventral part of the PVN. These results suggest that in spinally projecting PVN neurons, iNOS mainly contributes to the centrally administered CRF-induced activation of the sympatho-adrenomedullary outflow in rats. PMID- 19307159 TI - Adsorption and two-dimensional condensation of 5-methylcytosine. AB - Purine and pyrimidine derivatives occurring in nucleic acids posses an extraordinary high ability of self-association at the electrode surface and can form there by a two-dimensional (2D) condensation a monomolecular compact film (self-assembled monolayer-SAM). The effects of methyl substituent on the 2D condensation were studied using the 5-methylcytosine molecule which is involved in gene silencing and has a great biological impact. At acid pHs, 5 methylcytosine forms at the mercury electrode a physisorbed self-assembled 2D layer at potentials close to the potential of electrocapillary maximum. From the temperature dependence of the electrode double layer capacitance, the standard Gibbs energy of adsorption (Delta G(m)=-12.7 kJ mol(-1)), lateral interaction coefficient of the Frumkin adsorption isotherm (a(c)=2.05) and area occupied by one molecule (A=1.31 nm(2)) in the 2D layer were determined. Measurements performed on a single-crystal Au(111) surface show that the 2D condensation can take place on other substrates as well. PMID- 19307160 TI - Determination of procyanidins and their metabolites in plasma samples by improved liquid chromatography-tandem mass spectrometry. AB - An off-line solid-phase extraction (SPE) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for determining procyanidins, catechin, epicatechin, dimer, and trimer in plasma samples. In the validation procedure of the analytical method, linearity, precision, accuracy, detection limits (LODs), quantification limits (LOQs), and the matrix effect were studied. Recoveries of the procyanidins were higher than 84%, except for the trimer, which was 65%. The LODs and LOQs were lower than 0.003 and 0.01 microM, respectively, for all the procyanidins studied, except for the trimers, which were 0.8 and 0.98 microM, respectively. This methodology was then applied for the analysis of rat plasma obtained 2h after ingestion of grape seed phenolic extract. Monomers (catechin and epicatechin), dimer and trimer in their native form were detected and quantified in plasma samples, and their concentration ranged from 0.85 to 8.55 microM. Moreover, several metabolites, such as catechin and epicatechin glucuronide, catechin and epicatechin methyl glucuronide, and catechin and epicatechin methyl-sulphate were identified. These conjugated forms were quantified, in reference to the respective unconjugated form, showing concentrations between 0.06 and 23.90 microM. PMID- 19307161 TI - Effect of a matrix metalloproteinase sequestering biomaterial on Caco-2 epithelial cell barrier integrity in vitro. AB - A novel matrix metalloproteinase sequestering biomaterial (MI Theramer beads) restored the epithelial barrier in a double chamber in vitro test system after disruption by Cytochalasin D and the secretion of the metalloproteinase MMP-2. MI beads are chemically modified (hydroxamated) poly(methacrylic acid-co-methyl methacrylate). We are exploring the utility of this material in inflammatory bowel disease (IBD), in which one manifestation is a compromised intestinal epithelial barrier. In a first step towards this goal we incubated MI beads (or polymethyl methacrylate control beads) with Caco-2 epithelial cells and mesenchymal 3T3 fibroblasts on two sides of a Matrigel((R))-coated filter and used Cytochalasin D (Cyto D) to activate MMP-2 (secreted by the 3T3 cells), disrupt actin filaments of Caco-2 cells and render the epithelial barrier leaky, as measured by dextran fluorescein equilibration. Addition of MI beads to Cyto D treated cells inhibited active MMP-2 and prevented equilibration of dextran fluorescein. This study is the first step in showing a potential benefit to local (as opposed to systemic) inhibition of metalloproteinases in IBD or other intestinal inflammatory diseases. PMID- 19307162 TI - The importance of particle size in porous titanium and nonporous counterparts for surface energy and its impact on apatite formation. AB - The importance of particle size in titanium (Ti) fabricated by powder metallurgy for the surface energy and its impact on the apatite formation was investigated. Four sorts of Ti powders of different mean particle size were realized through 20min, 2h, 5h and 8h of ball milling, respectively. Each sort of Ti powder was used to fabricate porous Ti and its nonporous counterparts sharing similar surface morphology, grain size and chemical composition, and then alkali-heat treatment was conducted on them. Surface energy was measured on the surfaces of the nonporous Ti counterparts due to the difficulty in measuring the porous surfaces directly. The surface energy increase on the alkali-heat-treated porous and nonporous Ti was observed due to the decrease in the particle size of the Ti powders and the presence of Ti-OH groups brought by the alkali-heat treatment. The apatite-inducing ability of the alkali-heat-treated porous and nonporous Ti with different surface energy values was evaluated in modified simulated body fluid and results indicated that there was a strong correlation between the apatite-inducing ability and the surface energy. The alkali-heat-treated porous and nonporous Ti discs prepared from the powders with an average particle size of 5.89+/-0.76microm possessed the highest surface energy and the best apatite inducing ability when compared to the samples produced from the powders with the average particle size varying from 19.79+/-0.31 to 10.25+/-0.39microm. PMID- 19307163 TI - Modeling the effects of flow dispersion in arterial spin labeling. AB - Recent experimental results have shown that effects such as dispersion and cardiac pulsation have a significant effect on the arterial spin labeling (ASL) signal. These have not been incorporated into the existing ASL models potentially leading to inaccuracies in flow calculation. In this study, we develop a new model, based on physical principles, to model the transit of the ASL signal from the tagging band to the imaging band using the mass transport equation. We relax the assumption of a uniform plug flow, and account for the dispersion caused by the viscous nature of blood. The model also provides a framework within which other physiological aspects can easily be examined. Here, we examine the effects of flow dispersion on the ASL signal, and hence the quantification of cerebral perfusion. Our results suggest that not accounting for flow dispersion may result in inaccurate values of cerebral perfusion. PMID- 19307164 TI - A new approach to analysis and modeling of esophageal manometry data in humans. AB - In this paper, we propose a new approach to the analysis and modeling of esophageal manometry (EGM) data to assist the diagnosis of esophageal motility disorders in humans. The proposed approach combines three techniques, namely, wavelet decomposition (WD), nonlinear pulse detection technique (NPDT), and statistical pulse modeling. Specifically, WD is applied to the filtering of the EGM data, which is contaminated with electrocardiography (ECG) artifacts. A new NPDT is applied to the denoised data leading to identification and extraction of diagnostically important information, i.e., esophageal pulses from the respiration artifacts. Such information is used to generate a statistical model that can classify the EGM patterns. The proposed approach is computationally effortless, thus making it suitable for real-time application. Experimental results using measured EGM data of 20 patients, including ten abnormal cases is presented. Comparison of our results with those from existing techniques illustrates the advantages of the proposed approach in terms of accuracy and efficiency. PMID- 19307165 TI - Multivariate decomposition of arterial blood pressure variability for the assessment of arterial control of circulation. AB - In order to analyze the information carried by arterial blood pressure (ABP) variability, a multivariate parametric model of interactions involving systolic ABP (SAP), diastolic ABP (DAP), pulse pressure (PP), heart period (HP), and respiration is proposed. The model defines SAP as sum of the preceding DAP and PP values; DAP model accounts for arterial baroreflex, diastolic runoff; PP reflects changes in stroke volume related to respiration and HP, afterload; equation residuals reveal other vascular and cardiac output modulations. The model was applied to data from nine young volunteers (aged 29 +/- 6 years) during supine cycling at 10%, 20%, and 30% of their maximum effort. Significant basal values and changes across the epochs of the experiment were found in all hemodynamic parameters describing fast, beat-by-beat responses; in SAP and PP total power, DAP low- and high-frequency power (LF, HF), PP very low frequency (VLF), and LF and HF power. A primary role of vascular control through DAP and PP was emphasized by the considered feedbacks and the model residuals. The model proved to be able to assess beat-by-beat cardiovascular interactions and offer a comprehensive view of arterial tree control. PMID- 19307166 TI - Protein superfamily classification using fuzzy rule-based classifier. AB - In this paper, we have proposed a fuzzy rule-based classifier for assigning amino acid sequences into different superfamilies of proteins. While the most popular methods for protein classification rely on sequence alignment, our approach is alignment-free and so more human readable. It accounts for the distribution of contiguous patterns of n amino acids ( n-grams) in the sequences as features, alike other alignment-independent methods. Our approach, first extracts a plenty of features from a set of training sequences, then selects only some best of them, using a proposed feature ranking method. Thereafter, using these features, a novel steady-state genetic algorithm for extracting fuzzy classification rules from data is used to generate a compact set of interpretable fuzzy rules. The generated rules are simple and human understandable. So, the biologists can utilize them, for classification purposes, or incorporate their expertise to interpret or even modify them. To evaluate the performance of our fuzzy rule based classifier, we have compared it with the conventional nonfuzzy C4.5 algorithm, beside some other fuzzy classifiers. This comparative study is conducted through classifying the protein sequences of five superfamily classes, downloaded from a public domain database. The obtained results show that the generated fuzzy rules are more interpretable, with acceptable improvement in the classification accuracy. PMID- 19307167 TI - Multidetector CT of pancreatic hemangiopericytoma. AB - Hemangiopericytoma is an aggressive, highly metastatic tumor of the soft tissues and meninges. Metastases have been reported in the liver, lungs, bones and rarely other organs. To the best of our knowledge, 18 cases of pancreatic metastases have been reported, but none have been described in the radiology literature. We report a case of hemangiopericytoma with metastasis to the pancreas and describe the CT imaging features of hemangiopericytoma with a review of the literature. PMID- 19307168 TI - Identification of genomic predictors of non-melanoma skin cancer in solid organ transplant recipients. PMID- 19307169 TI - Be aware of proctitis and lymphogranuloma venereum. PMID- 19307170 TI - MLST typing of Escherichia coli isolates overproducing AmpC {beta}-lactamase. PMID- 19307171 TI - Influence of different resistance traits on the competitive growth of Haemophilus influenzae in antibiotic-free medium and selection of resistant populations by different {beta}-lactams: an in vitro pharmacodynamic approach. AB - OBJECTIVES: The aim was to study the pharmacodynamics of cefditoren, amoxicillin/clavulanic acid and cefuroxime against mixed Haemophilus influenzae strains. METHODS: Isolates [MICs (mg/L) of cefditoren, cefuroxime and amoxicillin/clavulanic acid] used were: one beta-lactamase-negative (beta(-); 0.015, 1 and 1), one beta-lactamase-positive (beta(+); 0.03, 4 and 8) and two strains exhibiting ftsI gene mutations [one beta(-) ampicillin-resistant (BLNAR; 0.015, 8 and 4) and one beta(+) amoxicillin/clavulanic acid-resistant (BLPACR; 0.03, 8 and 4)]. A computerized pharmacodynamic model simulating free antibiotic concentrations (calculated considering reported percentages of protein binding) of 400 mg twice-daily cefditoren, 500 mg twice-daily cefuroxime and 875/125 mg three times daily amoxicillin/clavulanic acid was used to explore antibacterial activity against initial mixed inocula with 25% of each strain. Areas under bacterial curves (AUBCs) from 0 to 24 h (log cfu.h/mL) were calculated and differences between values in antibiotic-free (AUBC(K)) and in antibiotic simulations determined (ABBC(0-24) = AUBC(K0-24)-AUBC(0-24)). RESULTS: In antibiotic-free medium, total population increased by 1.7 log(10) cfu/mL from 0 to 24 h: final composition approximately 90% beta(-), approximately 6.5% beta(+), approximately 2.5% BLNAR and approximately 1% BLPACR. At the end of antibiotic simulations, the predominant population was BLPACR followed by beta(+) after amoxicillin/clavulanic acid or BLNAR after cefuroxime exposures. ABBC(0-24) was higher (P < 0.01) for cefditoren compared with cefuroxime or amoxicillin/clavulanic acid whether considering total population (70.4 versus approximately 33), beta(+) (77.8 versus approximately 52), BLNAR (66.1 versus 18.6-30.4) or BLPACR (40.8 versus approximately 0). CONCLUSIONS: Cefditoren offered higher antibacterial effect than cefuroxime and amoxicillin/clavulanic acid against a mixed population of H. influenzae strains due to its higher activity against beta-lactamase-producing strains and those carrying ftsI gene mutations. PMID- 19307172 TI - Disruption of D-alanyl esterification of Staphylococcus aureus cell wall teichoic acid by the {beta}-lactam resistance modifier (-)-epicatechin gallate. AB - OBJECTIVES: The naturally occurring polyphenol (-)-epicatechin gallate (ECg) increases oxacillin susceptibility in mecA-containing strains of Staphylococcus aureus. Decreased susceptibility to lysostaphin suggests alterations to the wall teichoic acid (WTA) content of ECg-grown bacteria. Changes in WTA structure in response to ECg were determined. METHODS: Nuclear magnetic resonance spectroscopy of purified monomers from S. aureus was used to elucidate WTA structures. Molecular modelling of WTA chains was employed to determine their spatial configuration. RESULTS: ECg-grown methicillin-resistant S. aureus (MRSA) strains BB568 and EMRSA-16 displayed markedly reduced resistance to oxacillin, had thickened cell walls and separated poorly. Growth in ECg-supplemented medium reduced the substitution of the WTA backbone by d-alanine (d-Ala); ratios of N acetyl glucosamine to d-Ala were reduced from 0.6 and 0.49 (for BB568 and EMRSA 16) to 0.3 and 0.28, respectively. Molecular simulations indicated a decrease in the positive charge of the bacterial wall, confirmed by increased binding of cationized ferritin, and an increase in WTA chain flexibility to a random coil conformation. CONCLUSIONS: Structural elucidation and molecular modelling of WTA indicated that conformational changes associated with reduced d-Ala substitution may contribute to the increased susceptibility of MRSA to beta-lactam antibiotics and account for other elements of the ECg-induced phenotype. PMID- 19307173 TI - Gingyo-san enhances immunity and potentiates infectious bursal disease vaccination. AB - The purpose of the present study was to investigate the effects of Gingyo-san (GGS), a traditional Chinese medical formula, on peripheral lymphocyte proliferation and serum antibody titers in chickens vaccinated against the infectious bursal disease (IBD) virus. Treatment groups were fed one of three doses of GGS in their diet (0.5%, 1.0% and 2.0%, w/w), and the IBD vaccine was administered at 1 and 3 weeks of age. At Weeks 8, 12 and 16, changes in serum IBD antibody titers were measured via the micro-method and T cell proliferation. In gene expression experiments, GGS-treated peripheral T lymphocytes were stimulated with concanavalin A (ConA) for 24 h. The mRNA expression of interleukin-2 (IL-2), interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and interleukin-12 (IL-12) was determined using a semi-quantitative RT-PCR assay. The results showed that a low dose of GGS could significantly raise the antibody titers. Medium and high doses of GGS enhanced IL-2 and IFN-gamma production. GGS altered the expression of IL-4 and IL-12 in T lymphocytes. CD4(+) T lymphocyte development was also skewed towards the Th1 phenotype. GGS enhanced cell-mediated immunity and augmented the effects of IBD vaccination in strengthening subsequent anti-viral responses. PMID- 19307174 TI - A unified scheme for initiation and conformational adaptation of human apolipoprotein E N-terminal domain upon lipoprotein binding and for receptor binding activity. AB - We report here a high-resolution NMR structure of the complete receptor-binding domain of human apolipoprotein E3 (apoE3-NT). Similar to the crystal structure of apoE-NT, the NMR structure displayed an elongated four-helix bundle. However, additional unique structural features were also observed. The segments in the N and C termini, which were missing in the crystal structure, formed alpha-helices having extensive tertiary contacts with the bundle, which oriented these short helices at specific positions for receptor binding activity. Several buried hydrophilic residues observed in the bundle were located strategically between helices 1 and 2 and between helices 3 and 4, significantly destabilizing these helix-helix interfaces. In addition, these buried hydrophilic residues formed buried H-bonds, which may play a key role in specific lipid-free helix bundle recovery. A short helix, nHelix C, was fully solvent-exposed and nearly perpendicular to the bundle. This short helix likely plays a critical role in initiating protein-lipid interaction, causing a preferred conformational adaptation of the bundle at the weaker helix-helix interfaces. This produces an open conformation with two lobes of helices, helices 1 and 4 and helices 2 and 3, which may be the competent conformation for receptor binding activity. Thus, the NMR structure suggests a unified scheme for the initiation and helix bundle opening of apoE-NT upon lipoprotein-binding and for receptor binding activity. PMID- 19307175 TI - Dissection of mechanisms involved in the regulation of Plasmodium falciparum calcium-dependent protein kinase 4. AB - Recent studies have demonstrated that calcium-dependent protein kinases (CDPKs) are used by calcium to regulate a variety of biological processes in the malaria parasite Plasmodium. CDPK4 has emerged as an important enzyme for parasite development, because its gene disruption in rodent parasite Plasmodium berghei causes major defects in sexual differentiation of the parasite ( Billker, O., Dechamps, S., Tewari, R., Wenig, G., Franke-Fayard, B., and Brinkmann, V. (2004) Cell 117, 503-514 ). Despite these findings, it is not very clear how PfCDPK4 or any other PfCDPK is regulated by calcium at the molecular level. We report the biochemical characterization and elucidation of molecular mechanisms involved in the regulation of PfCDPK4. PfCDPK4 was detected on gametocyte periphery, and its activity in the parasite was regulated by phospholipase C. Even though the Junction Domain (JD) of PfCDPK4 shares moderate sequence homology with that of the plant CDPKs, it plays a pivotal role in PfCDPK4 regulation as previously reported for some plant CDPKs. The regions of the J-domain involved in interaction with both the kinase domain and the calmodulin-like domain were mapped. We propose a model for PfCDPK regulation by calcium, which may also prove useful for design of inhibitors against PfCDPK4 and other members of the PfCDPK family. PMID- 19307176 TI - Coexpression and heteromerization of two neuronal K-Cl cotransporter isoforms in neonatal brain. AB - The neuron-specific K-Cl cotransporter KCC2 maintains the low intracellular chloride concentration required for the fast hyperpolarizing actions of inhibitory neurotransmitters. The KCC2 gene codes for two isoforms, KCC2a and KCC2b, which differ in their N termini. The relative expression and cellular distribution of the two KCC2 protein isoforms are unknown. Here, we characterize an antibody against the KCC2a isoform and show that a previously described antibody against KCC2 is specific for the KCC2b isoform (Hubner, C. A., Stein, V., Hermans-Borgmeyer, I., Meyer, T., Ballanyi, K., and Jentsch, T. J. (2001) Neuron 30, 515-524). Immunostaining of dissociated hippocampal cultures confirms that both KCC2 isoforms are neuron-specific. Immunoblot analysis indicates that KCC2b is the major KCC2 isoform in the adult brain, whereas in the neonatal mouse central nervous system, half of total KCC2 protein is KCC2a. At this stage, the two KCC2 isoforms are largely colocalized and show similar patterns of distribution in the brain. When coexpressed in HEK293 cells, KCC2a and KCC2b proteins form heteromeric complexes. Moreover, the two isoforms can be coimmunoprecipitated from the neonatal brain, suggesting the presence of endogenous KCC2a-KCC2b heteromers. Consistent with this, native gel analysis shows that a substantial part of endogenous KCC2 isoforms in the neonatal brain constitute dimers. PMID- 19307177 TI - Use of the pharmacological inhibitor BX795 to study the regulation and physiological roles of TBK1 and IkappaB kinase epsilon: a distinct upstream kinase mediates Ser-172 phosphorylation and activation. AB - TANK-binding kinase 1 (TBK1) and IkappaB kinase epsilon (IKKepsilon) regulate the production of Type 1 interferons during bacterial and viral infection, but the lack of useful pharmacological inhibitors has hampered progress in identifying additional physiological roles of these protein kinases and how they are regulated. Here we demonstrate that BX795, a potent and relatively specific inhibitor of TBK1 and IKKepsilon, blocked the phosphorylation, nuclear translocation, and transcriptional activity of interferon regulatory factor 3 and, hence, the production of interferon-beta in macrophages stimulated with poly(I:C) or lipopolysaccharide (LPS). In contrast, BX795 had no effect on the canonical NFkappaB signaling pathway. Although BX795 blocked the autophosphorylation of overexpressed TBK1 and IKKepsilon at Ser-172 and, hence, the autoactivation of these protein kinases, it did not inhibit the phosphorylation of endogenous TBK1 and IKKepsilon at Ser-172 in response to LPS, poly(I:C), interleukin-1alpha (IL-1alpha), or tumor necrosis factor alpha and actually enhanced the LPS, poly(I:C), and IL-1alpha-stimulated phosphorylation of this residue. These results demonstrate that the phosphorylation of Ser-172 and the activation of TBK1 and IKKepsilon are catalyzed by a distinct protein kinase(s) in vivo and that TBK1 and IKKepsilon control a feedback loop that limits their activation by LPS, poly(I:C) and IL-1alpha (but not tumor necrosis factor alpha) to prevent the hyperactivation of these enzymes. PMID- 19307178 TI - Engineered human antibody constant domains with increased stability. AB - The immunoglobulin (Ig) constant CH2 domain is critical for antibody effector functions. Isolated CH2 domains are promising as scaffolds for construction of libraries containing diverse binders that could also confer some effector functions. However, previous work has shown that an isolated murine CH2 domain is relatively unstable to thermally induced unfolding. To explore unfolding mechanisms of isolated human CH2 and increase its stability gamma1 CH2 was cloned and a panel of cysteine mutants was constructed. Human gamma1 CH2 unfolded at a higher temperature (T(m) = 54.1 degrees C, as measured by circular dichroism) than that previously reported for a mouse CH2 (41 degrees C). One mutant (m01) was remarkably stable (T(m) = 73.8 degrees C). Similar results were obtained by differential scanning calorimetry. This mutant was also significantly more stable than the wild-type CH2 against urea induced unfolding (50% unfolding at urea concentration of 6.8 m versus 4.2 m). The m01 was highly soluble and monomeric. The existence of the second disulfide bond in m01 and its correct position were demonstrated by mass spectrometry and nuclear magnetic resonance spectroscopy, respectively. The loops were on average more flexible than the framework in both CH2 and m01, and the overall secondary structure was not affected by the additional disulfide bond. These data suggest that a human CH2 domain is relatively stable to unfolding at physiological temperature, and that both CH2 and the highly stable mutant m01 are promising new scaffolds for the development of therapeutics against human diseases. PMID- 19307179 TI - A promoter recognition mechanism common to yeast mitochondrial and phage t7 RNA polymerases. AB - Yeast mitochondrial (YMt) and phage T7 RNA polymerases (RNAPs) are two divergent representatives of a large family of single subunit RNAPs that are also found in the mitochondria and chloroplasts of higher eukaryotes, mammalian nuclei, and many other bacteriophage. YMt and phage T7 promoters differ greatly in sequence and length, and the YMt RNAP uses an accessory factor for initiation, whereas T7 RNAP does not. We obtain evidence here that, despite these apparent differences, both the YMt and T7 RNAPs utilize a similar promoter recognition loop to bind their respective promoters. Mutations in this element in YMt RNAP specifically disrupt mitochondrial promoter utilization, and experiments with site specifically tethered chemical nucleases indicate that this element binds the mitochondrial promoter almost identically to how the promoter recognition loop from the phage RNAP binds its promoter. Sequence comparisons reveal that the other members of the single subunit RNAP family display loops of variable sequence and size at a position corresponding to the YMt and T7 RNAP promoter recognition loops. We speculate that these elements may be involved in promoter recognition in most or all of these enzymes and that this element's structure allows it to accommodate significant sequence and length variation to provide a mechanism for rapid evolution of new promoter specificities in this RNAP family. PMID- 19307180 TI - The Ste20 kinases Ste20-related proline-alanine-rich kinase and oxidative-stress response 1 regulate NKCC1 function in sensory neurons. AB - NKCC1 is highly expressed in dorsal root ganglion neurons, where it is involved in gating sensory information. In a recent study, it was shown that peripheral nerve injury results in increased NKCC1 activity, not due to an increase in cotransporter expression, but to increased phosphorylation of the cotransporter (Pieraut, S., Matha, V., Sar, C., Hubert, T., Mechaly, I., Hilaire, C., Mersel, M., Delpire, E., Valmier, J., and Scamps, F. (2007) J. Neurosci. 27, 6751-6759). Our laboratory has also identified two Ste20-like kinases that bind and phosphorylate NKCC1: Ste20-related proline-alanine-rich kinase (SPAK) and oxidative-stress response 1 (OSR1). In this study, we show that both kinases are expressed at similar expression levels in spinal cord and dorsal root ganglion neurons, and that both kinases participate equally in the regulation of NKCC1. Using a novel fluorescence method to assay NKCC1 activity in single cells, we show a 50% reduction in NKCC1 activity in DRG neurons isolated from SPAK knockout mice, indicating that another kinase, e.g. OSR1, is present to phosphorylate and activate the cotransporter. Using a nociceptive dorsal root ganglion sensory neuronal cell line, which expresses the same cation-chloride cotransporters and kinases as native DRG neurons, and gene silencing via short hairpin RNA, we demonstrate a direct relationship between kinase expression and cotransporter activity. We show that inactivation of either kinase significantly affects NKCC1 activity, whereas inactivation of both kinases results in an additive effect. In summary, our study demonstrates redundancy of kinases in the regulation of NKCC1 in dorsal root ganglion neurons. PMID- 19307181 TI - Acylated cholesteryl galactosides are specific antigens of borrelia causing lyme disease and frequently induce antibodies in late stages of disease. AB - Borrelia burgdorferi sensu lato is the causative agent of Lyme disease (LD), an infectious disease occurring in North America, Europe, and Asia in different clinical stages. B. burgdorferi sensu lato encompasses at least 12 species, with B. burgdorferi sensu stricto, B. garinii, and B. afzelii being of highest clinical importance. Immunologic testing for LD as well as recent vaccination strategies exclusively refer to proteinaceous antigens. However, B. burgdorferi sensu stricto exhibits glycolipid antigens, including 6-O-acylated cholesteryl beta-D-galactopyranoside (ACGal), and first the data indicated that this compound may act as an immunogen. Here we investigated whether B. garinii and B. afzelii also possess this antigen, and whether antibodies directed against these compounds are abundant among patients suffering from different stages of LD. Gas liquid chromatography/mass spectroscopy and NMR spectroscopy showed that both B. garinii and B. afzelii exhibit ACGal in high quantities. In contrast, B. hermsii causing relapsing fever features 6-O-acylated cholesteryl beta-D-glucopyranoside (ACGlc). Sera derived from patients diagnosed for LD contained antibodies against ACGal, with 80% of patients suffering from late stage disease exhibiting this feature. Antibodies reacted with ACGal from all three B. burgdorferi species tested, but not with ACGlc from B. hermsii. These data show that ACGal is present in all clinically important B. burgdorferi species, and that specific antibodies against this compound are frequently found during LD. ACGal may thus be an interesting tool for improving diagnostics as well as for novel vaccination strategies. PMID- 19307182 TI - Recepteur d'origine nantais tyrosine kinase is a direct target of hypoxia inducible factor-1alpha-mediated invasion of breast carcinoma cells. AB - Hypoxia-inducible factor-1alpha (HIF-1alpha) overexpression was shown to be associated with invasion and metastasis of tumors and tumor cell lines. The identification of molecular targets that contribute to HIF-1alpha-mediated invasion is under intensive investigation. We have analyzed the role of recepteur d'origine nantais (RON), a tyrosine kinase receptor for macrophage-stimulating protein (MSP) that plays a role in breast cancer cell invasion as one of the molecular targets of HIF-1alpha. Analysis of a panel of breast cancer cell lines indicated a correlation between HIF-1alpha and RON expression. Treatment of HIF 1alpha- and RON-positive breast cancer cells with HIF-1alpha inhibitor, echinomycin, led to the inhibition of HIF-1alpha activity and RON expression. We have identified HIF-1alpha binding site on the RON promoter. Chromatin immunoprecipitation analysis and site-directed mutagenesis of the RON promoter confirmed the binding of HIF-1alpha to RON promoter. HIF-1alpha inhibitor-, echinomycin-, or short hairpin RNA-mediated selective knockdown of HIF-1alpha or HIF-1alpha target RON tyrosine kinase abrogated RON gene expression, and the RON ligand macrophage-stimulating protein mediated invasion of breast cancer cells. Consequently, the data presented herein demonstrated RON as a novel molecular target of HIF-1alpha and suggest a potential therapeutic role for HIF-1alpha or RON tyrosine kinase inhibitors in the blockade of RON tyrosine kinase-mediated invasion of carcinoma cells. PMID- 19307183 TI - Light-induced dissociation of an antenna hetero-oligomer is needed for non photochemical quenching induction. AB - PsbS plays a major role in activating the photoprotection mechanism known as "non photochemical quenching," which dissipates chlorophyll excited states exceeding the capacity for photosynthetic electron transport. PsbS activity is known to be triggered by low lumenal pH. However, the molecular mechanism by which this subunit regulates light harvesting efficiency is still unknown. Here we show that PsbS controls the association/dissociation of a five-subunit membrane complex, composed of two monomeric Lhcb proteins (CP29 and CP24) and the trimeric LHCII-M. Dissociation of this supercomplex is indispensable for the onset of non photochemical fluorescence quenching in high light, strongly suggesting that protein subunits catalyzing the reaction of heat dissipation are buried into the complex and thus not available for interaction with PsbS. Consistently, we showed that knock-out mutants on two subunits participating to the B4C complex were strongly affected in heat dissipation. Direct observation by electron microscopy and image analysis showed that B4C dissociation leads to the redistribution of PSII within grana membranes. We interpreted these results to mean that the dissociation of B4C makes quenching sites, possibly CP29 and CP24, available for the switch to an energy-quenching conformation. These changes are reversible and do not require protein synthesis/degradation, thus allowing for changes in PSII antenna size and adaptation to rapidly changing environmental conditions. PMID- 19307184 TI - Phosphatidylinositol 3-kinase-dependent activation of mammalian protein kinase B/Akt in Saccharomyces cerevisiae, an in vivo model for the functional study of Akt mutations. AB - In animal cells, Akt (also called protein kinase B) is activated by stimuli that elevate the level of phosphatidylinositol 3,4,5-trisphosphate and is a major effector for eliciting responses that support cell growth and survival. We have shown previously that co-expression of Akt1 in budding yeast (Saccharomyces cerevisiae) along with hyperactive p110alpha, the catalytic subunit of mammalian phosphatidylinositol 3-kinase, results in Akt1 relocalization to cellular membranes and activation. In the present study, we show that activation of all three mammalian Akt isoforms by wild-type p110alpha causes deleterious effects on yeast cell growth. Toxicity of Akt in S. cerevisiae required its catalytic activity, its pleckstrin homology domain, and phosphorylation of its activation loop, but not phosphorylation of its hydrophobic motif. We demonstrate that expression in yeast of the only purported oncogenic allele, Akt1(E17K), leads to enhanced phenotypes. Ala-scanning mutagenesis of the VL1 region within the phosphatidylinositol 3,4,5-trisphosphate-interacting pocket of the Akt1 pleckstrin homology domain revealed that most residues in this region are essential for Akt1 activity. We found that active Akt leads to enhanced signaling through the yeast cell wall integrity pathway. This effect requires the upstream Rho1 activator Rom2 and involves both phosphorylation of the MAPK Slt2 and expression of its transcriptional targets, thus providing a quantitative reporter system for heterologous Akt activity in vivo. Collectively, our results disclose a heterologous yeast system that allows the functional assessment in vivo of both loss-of-function and tumorigenic Akt alleles. PMID- 19307185 TI - Cellular functions and X-ray structure of anthrolysin O, a cholesterol-dependent cytolysin secreted by Bacillus anthracis. AB - Anthrolysin O (ALO) is a pore-forming, cholesterol-dependent cytolysin (CDC) secreted by Bacillus anthracis, the etiologic agent for anthrax. Growing evidence suggests the involvement of ALO in anthrax pathogenesis. Here, we show that the apical application of ALO decreases the barrier function of human polarized epithelial cells as well as increases intracellular calcium and the internalization of the tight junction protein occludin. Using pharmacological agents, we also found that barrier function disruption requires increased intracellular calcium and protein degradation. We also report a crystal structure of the soluble state of ALO. Based on our analytical ultracentrifugation and light scattering studies, ALO exists as a monomer. Our ALO structure provides the molecular basis as to how ALO is locked in a monomeric state, in contrast to other CDCs that undergo antiparallel dimerization or higher order oligomerization in solution. ALO has four domains and is globally similar to perfringolysin O (PFO) and intermedilysin (ILY), yet the highly conserved undecapeptide region in domain 4 (D4) adopts a completely different conformation in all three CDCs. Consistent with the differences within D4 and at the D2-D4 interface, we found that ALO D4 plays a key role in affecting the barrier function of C2BBE cells, whereas PFO domain 4 cannot substitute for this role. Novel structural elements and unique cellular functions of ALO revealed by our studies provide new insight into the molecular basis for the diverse nature of the CDC family. PMID- 19307186 TI - Angiotensin-converting enzyme is a modifier of hypertensive end organ damage. AB - Severe forms of hypertension are characterized by high blood pressure combined with end organ damage. Through the development and refinement of a transgenic rat model of malignant hypertension incorporating the mouse renin gene, we previously identified a quantitative trait locus on chromosome 10, which affects malignant hypertension severity and morbidity. We next generated an inducible malignant hypertensive model where the timing, severity, and duration of hypertension was placed under the control of the researcher, allowing development of and recovery from end organ damage to be investigated. We have now generated novel consomic Lewis and Fischer rat strains with inducible hypertension and additional strains that are reciprocally congenic for the refined chromosome 10 quantitative trait locus. We have captured a modifier of end organ damage within the congenic region and, using a range of bioinformatic, biochemical and molecular biological techniques, have identified angiotensin-converting enzyme as the modifier of hypertension-induced tissue microvascular injury. Reciprocal differences between angiotensin-converting enzyme and the anti-inflammatory tetrapeptide, N-acetyl Ser-Asp-Lys-Pro in the kidney, a tissue susceptible to end organ damage, suggest a mechanism for the amelioration of hypertension-dependent damage. PMID- 19307187 TI - Interleukin-6 released from fibroblasts is essential for up-regulation of matrix metalloproteinase-1 expression by U937 macrophages in coculture: cross-talking between fibroblasts and U937 macrophages exposed to high glucose. AB - Matrix metalloproteinases (MMPs) play a key role in periodontal disease. Although it is known that macrophages and fibroblasts are co-localized and express MMPs in the diseased periodontal tissue, the effect of interaction between these two cell types on MMP expression has not been well elucidated. Furthermore although it is known that diabetes is associated with accelerated periodontal tissue destruction, it remains unknown whether hyperglycemia, a major metabolic abnormality in diabetes, regulates MMP expression by affecting the cross-talking between fibroblasts and macrophages. In this study, human gingival fibroblasts and U937 macrophages were cocultured in a two-compartment transwell culture system, and the cells were treated with normal or high glucose. We found that coculture of fibroblasts and U937 macrophages led to an augmentation of MMP-1 expression by U937 macrophages, and high glucose further enhanced this augmentation. Similar observations were also made in the coculture of fibroblasts and human primary monocytes. We also found that interleukin 6 (IL-6) released by fibroblasts was essential for the augmentation of MMP-1 expression by U937 macrophages. Furthermore our results showed that high glucose, IL-6, and lipopolysaccharide had a synergistic effect on MMP-1 expression. Finally our study indicated that MAPK pathways and activator protein-1 transcription factor were involved in the coculture- and high glucose-augmented MMP-1 expression. In conclusion, this study demonstrates that IL-6 derived from fibroblasts is essential for MMP-1 up-regulation by cross-talking between fibroblasts and U937 macrophages exposed to high glucose, revealing an IL-6-dependent mechanism in MMP 1 up-regulation. PMID- 19307188 TI - Mutants of the Arabidopsis thaliana cation/H+ antiporter AtNHX1 conferring increased salt tolerance in yeast: the endosome/prevacuolar compartment is a target for salt toxicity. AB - Mutants of the plant cation/H(+) antiporter AtNHX1 that confer greater halotolerance were generated by random mutagenesis and selected in yeast by phenotypic complementation. The amino acid substitutions that were selected were conservative and occurred in the second half of the membrane-associated N terminus. AtNHX1 complemented the lack of endogenous ScNHX1 in endosomal protein trafficking assays. Growth enhancement on hygromycin B and vanadate media agreed with a generally improved endosomal/prevacuolar function of the mutated proteins. In vivo measurements by (31)P NMR revealed that wild-type and mutant AtNHX1 transporters did not affect cytosolic or vacuolar pH. Surprisingly, when yeast cells were challenged with lithium, a tracer for sodium, the main effect of the mutations in AtNHX1 was a reduction in the amount of compartmentalized lithium. When purified and reconstituted into proteoliposomes or assayed in intact vacuoles isolated from yeast cells, a representative mutant transporter (V318I) showed a greater cation discrimination favoring potassium transport over that of sodium or lithium. Together, our data suggest that the endosome/prevacuolar compartment is a target for salt toxicity. Poisoning by toxic cations in the endosome/prevacuolar compartment is detrimental for cell functions, but it can be alleviated by improving the discrimination of transported alkali cations by the resident cation/H(+) antiporter. PMID- 19307189 TI - Effect of pitcher age on trapping efficiency and natural prey capture in carnivorous Nepenthes rafflesiana plants. AB - BACKGROUND AND AIMS: Nepenthes pitchers are sophisticated traps that employ a variety of mechanisms to attract, capture and retain prey. The underlying morphological structures and physiological processes are subject to change over the lifetime of a pitcher. Here an investigation was carried out on how pitcher properties and capture efficiency change over the first 2 weeks after pitcher opening. METHODS: Prey capture, trapping efficiency, extrafloral nectar secretion, pitcher odour, as well as pH and viscoelasticity of the digestive fluid in N. rafflesiana pitchers were monitored in the natural habitat from pitcher opening up to an age of 2 weeks. KEY RESULTS: Pitchers not only increased their attractiveness over this period by becoming more fragrant and secreting more nectar, but also gained mechanical trapping efficiency via an enhanced wettability of the upper pitcher rim (peristome). Consistently, natural prey capture was initially low and increased 3-6 d after opening. It was, however, highly variable within and among pitchers. At the same time, the pH and viscoelasticity of the digestive fluid decreased, suggesting that the latter is not essential for effective prey capture. CONCLUSIONS: Prey capture and attraction by Nepenthes are dynamic processes strongly influenced by the changing properties of the pitcher. The results confirm insect aquaplaning on the peristome as the main capture mechanism in N. rafflesiana. PMID- 19307190 TI - Phenotypic, genetic and genomic consequences of natural and synthetic polyploidization of Nicotiana attenuata and Nicotiana obtusifolia. AB - BACKGROUND AND METHODS: Polyploidy results in genetic turmoil, much of which is associated with new phenotypes that result in speciation. Five independent lines of synthetic allotetraploid N. x obtusiata (N x o) were created from crosses between the diploid N. attenuata (Na) (male) and N. obtusifolia (No) (female) and the autotetraploids of Na (NaT) and No (NoT) were synthesized. Their genetic, genomic and phenotypic changes were then compared with those of the parental diploid species (Na and No) as well as to the natural allotetraploids, N. quadrivalvis (Nq) and N. clevelandii (Nc), which formed 1 million years ago from crosses between ancient Na and No. KEY RESULTS: DNA fingerprinting profiles (by UP-PCR) revealed that the five N x o lines shared similar but not identical profiles. Both synthetic and natural polyploidy showed a dosage effect on genome size (as measured in seeds); however, only Nq was associated with a genome upsizing. Phenotypic analysis revealed that at the cellular level, N x o lines had phenotypes intermediate of the parental phenotypes. Both allo- and autotetraploidization had a dosage effect on seed and dry biomass (except for NaT), but not on stalk height at first flower. Nc showed paternal (Na) cellular phenotypes but inherited maternal (No) biomass and seed mass, whereas Nq showed maternal (No) cellular phenotypes but inherited paternal (Na) biomass and seed mass patterns. Principal component analysis grouped Nq with N x o lines, due to similar seed mass, stalk height and genome size. These traits separated Nc, No and Na from Nq and N x o lines, whereas biomass distinguished Na from N x o and Nq lines, and NaT clustered closer to Nq and N x o lines than to Na. CONCLUSIONS: Both allo- and autotetraploidy induce considerable morphological, genetic and genomic changes, many of which are retained by at least one of the natural polyploids. It is proposed that both natural and synthetic polyploids are well suited for studying the evolution of adaptive responses. PMID- 19307191 TI - Gedankenexperiment or just a flight of fancy? PMID- 19307192 TI - Decreased L-ascorbate content mediating bolting is mainly regulated by the galacturonate pathway in Oncidium. AB - We investigated the alteration in l-ascorbate (AsA, reduced form) content and the expression pattern of its related genes during the phase transition in Oncidium orchid. During the vegetative growth, a high H2O2 level was associated with a high content of the reduced form of AsA. During the bolting period, the AsA content and H2O2 level were greatly reduced in parallel with increased expression of OgLEAFY, the gene encoding a key transcription factor integrating different flowering-inducing pathways. This observation suggests that reduced AsA content, due to it having been consumed in scavenging H2O2, is a prerequisite for mediating the phase transition in Oncidium. A survey of the AsA biosynthetic pathway revealed that the gene expression and enzymatic activities of the products of relevant genes of the galacturonate (GalUA) pathway, such as polygalacturonase (OgPG), pectin methylesterase (OgPME) and galacturonate reductase (OgGalUAR), were markedly decreased during the bolting period, as compared with during the vegetative stage. However, the genes whose products were involved in the Smirnoff-Wheeler pathway retained a similar expression level in the two growth stages. The data suggested that OgPME of the GalUA pathway was the pivotal gene in regulating AsA biosynthesis during the bolting period. Further elucidation by overexpressing OgPME in Arabidopsis demonstrated a considerable increase in AsA content, as well as a resulting delayed-flowering phenotype. Our results strongly imply that the reduced level of AsA, regulating bolting for phase transition, resulting in part from its consumption by scavenging H2O2, was mainly caused by the down-regulation of the GalUA pathway, not the Smirnoff Wheeler pathway. PMID- 19307193 TI - Give Hippocrates a jersey: promoting health through football/sport. PMID- 19307194 TI - Are adolescents with chronic conditions particularly at risk for bullying? AB - OBJECTIVE: To compare the prevalence and intensity of victimisation from bullying and the characteristics of the victim of bullying, comparing adolescents with and adolescents without chronic conditions (CC). DESIGN: School survey. SETTING: Postmandatory schools. PARTICIPANTS: A total of 7005 students (48% females) aged 16-20 years, distributed into adolescents with CC (728, 50% females) and controls (6277, 48% females). Chronic condition was defined as having a chronic disease and/or a physical disability. OUTCOME MEASURES: Prevalence of bullying-intensity of bullying-and sociodemographic, biopsychosocial, familial, school and violence context characteristics of the victims of bullying. RESULTS: The prevalence of bullying in our sample was 13.85%. Adolescents with CC were more likely to be victims of bullying (adjusted OR 1.53), and to be victims of two or three forms of bullying (adjusted OR 1.92). Victims of bullying with CC were more likely than non-victims to be depressed (RR 1.57), to have more physical symptoms (RR 1.61), to have a poorer relationship with their parents (RR 1.33), to have a poorer school climate (RR 1.60) and to have been victims of sexual abuse (RR 1.79) or other forms of violence (RR 1.80). Although these characteristics apply to victims in general, in most cases, they are less pronounced among victims without CC. CONCLUSIONS: CC seems to be a risk factor for victimisation from bullying. Therefore, as adolescents with CC are increasingly mainstreamed, schools should be encouraged to undertake preventive measures to avoid victimisation of such adolescents. PMID- 19307195 TI - Congenital valvular aortic stenosis: limited progression during childhood. AB - OBJECTIVE: Congenital valvular aortic stenosis is a common congenital heart malformation. The rate of progression in childhood, however, remains to be established. We assessed the progression of peak aortic velocity before intervention as well as the frequency of intervention in paediatric patients with isolated congenital valvular aortic stenosis. METHODS: A retrospective cohort study was performed in 245 consecutive patients with aortic stenosis. Both clinical and echocardiographic data were obtained. RESULTS: Over a period of 9.0 (SD 5.2) years (range 0.1-19.4), the mean annual increase in peak systolic velocity was 0.04 m/s/year (95% CI 0.028 to 0.056 m/s/year; p<0.001) as shown by ANOVA. 40 patients underwent a cardiac intervention shortly after their first echocardiogram. Another 33 patients underwent intervention during follow-up. Interventions were performed significantly more often in patients diagnosed at a younger age and/or with a higher peak velocity at diagnosis (p<0.001). Mortality was considerable in those diagnosed in infancy (5-year survival rate of 73% (SD 9%), whereas it was nearly absent in patients diagnosed after infancy. Most patients who died during infancy had progressive left ventricular dysfunction despite adequate relief of left ventricular outflow obstruction. CONCLUSIONS: Valvular aortic stenosis in the paediatric age group usually has a good prognosis beyond the neonatal period. Progression over time is usually limited, although a considerable proportion of patients need intervention shortly after initial diagnosis. Mortality, except for the neonatal age group, is nearly absent. PMID- 19307196 TI - Quality indicators and quality assessment in child health. AB - Quality indicators are systematically developed statements that can be used to assess the appropriateness of specific healthcare decisions, services and outcomes. In this review, the range and type of indicators that have been developed for children in the UK and USA by prominent governmental agencies and private organisations are highlighted. These indicators are classified in an effort to identify areas of child health that may lack quality measures. The current state of health information technology in both countries is reviewed, since these systems are vital to quality efforts. Finally, several recommendations are proposed to advance the quality indicator development agenda for children. The convergence of quality measurement and indicator development, a growing scientific evidence base and integrated information systems in healthcare may lead to substantial improvements for child health in the 21st century. PMID- 19307197 TI - Changing trends in antiepileptic drug prescribing in girls of child-bearing potential. AB - OBJECTIVE: To characterize trends in prescribing carbamazepine (CBZ), sodium valproate (VPA) and lamotrigine (LTG) in adolescent females in the UK and to examine possible reasons for changing trends. DESIGN: Population-based observational study. SETTING: UK General Practice Research Database between 1 January 1993 and 31 December 2006. PATIENTS: 12-18-year-old subjects who were issued >or=1 CBZ, VPA or LTG prescription. MAIN OUTCOME MEASURES: Prescribing prevalences stratified by age, gender and antiepileptic drug. RESULTS: 5417 patients (47.6% females) were prescribed 147 111 prescriptions for CBZ (34.5%), VPA (38.6%) or LTG (26.9%). The prevalence of LTG prescribing in females increased from 0.08 (95% CI 0.04 to 0.12) to 0.80 (95% CI 0.70 to 0.89) per 1000 female population. Conversely, the prevalence in females of CBZ and VPA prescribing significantly decreased from 1.00 (95% CI 0.85 to 1.15) to 0.51 (95% CI 0.44 to 0.58) and from 0.94 (95% CI 0.80 to 1.09) to 0.63 (95% CI 0.55 to 0.72), respectively. This 10-fold rise in LTG prescribing in females is much higher than the fivefold rise in males from 0.09 (95% CI 0.05 to 0.14) to 0.47 (95% CI 0.40 to 0.54) per 1000 male population. CONCLUSION: The practice of prescribing antiepileptic drugs in adolescents has changed gradually over the last decade. More females aged 12-18 years are prescribed LTG than CBZ or VPA and the increase is much greater than for males. The increase in LTG prescribing mirrors a corresponding decrease in both VPA and CBZ. Concerns about potential problems to offspring appear to be affecting prescription trends in adolescent females of child-bearing potential. PMID- 19307198 TI - Cardiac resynchronisation therapy in paediatric and congenital heart disease: differential effects in various anatomical and functional substrates. AB - BACKGROUND: Cardiac resynchronisation therapy (CRT) is increasingly used in children in a variety of anatomical and pathophysiological conditions, but published data are scarce. OBJECTIVE: To record current practice and results of CRT in paediatric and congenital heart disease. DESIGN: Retrospective multicentre European survey. SETTING: Paediatric cardiology and cardiac surgery centres. PATIENTS: One hundred and nine patients aged 0.24-73.8 (median 16.9) years with structural congenital heart disease (n = 87), congenital atrioventricular block (n = 12) and dilated cardiomyopathy (n = 10) with systemic left (n = 69), right (n = 36) or single (n = 4) ventricular dysfunction and ventricular dyssynchrony during sinus rhythm (n = 25) or associated with pacing (n = 84). INTERVENTIONS: CRT for a median period of 7.5 months (concurrent cardiac surgery in 16/109). MAIN OUTCOME MEASURES: Functional improvement and echocardiographic change in systemic ventricular function. RESULTS: The z score of the systemic ventricular end-diastolic dimension decreased by median 1.1 (p<0.001). Ejection fraction (EF) or fractional area of change increased by a mean (SD) of 11.5 (14.3)% (p<0.001) and New York Heart Association (NYHA) class improved by median 1.0 grade (p<0.001). Non-response to CRT (18.5%) was multivariably predicted by the presence of primary dilated cardiomyopathy (p = 0.002) and poor NYHA class (p = 0.003). Presence of a systemic left ventricle was the strongest multivariable predictor of improvement in EF/fractional area of change (p<0.001). Results were independent of the number of patients treated in each contributing centre. CONCLUSION: Heart failure associated with ventricular pacing is the largest indication for CRT in paediatric and congenital heart disease. CRT efficacy varies widely with the underlying anatomical and pathophysiological substrate. PMID- 19307199 TI - Diabetes in the young: what are their long term health prospects? PMID- 19307200 TI - Pitfalls in imaging subdural haematoma. PMID- 19307201 TI - An unusual case of wheeze. PMID- 19307202 TI - Successful treatment of anti-N-methyl-D-aspartate receptor encephalitis presenting with catatonia. AB - The case of a 12-year-old girl with the typical clinical symptoms of the recently described anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is reported. Within 6 weeks the full clinical spectrum of this condition presented with seizures, agitation, stupor, autonomic instability, dysphagia and hypoventilation leading to a diagnosis of pernicious catatonia. MRI and CSF glucose, protein and lactate were repeatedly normal. EEG revealed rhythmical slowing. No teratoma was detected. Recognition of the unique pattern of the clinical symptoms led to early consideration of this disease which was confirmed by detection of anti-NMDAR antibodies. After high dose prednisolone without clinical improvement, plasmapheresis was followed by a rapid reduction in antibodies and recovery within a few weeks. To our knowledge this is the youngest patient with anti-NMDAR encephalitis to have been described to date. We speculate that NMDAR antibodies may be directly involved in the pathogenesis of this disease. PMID- 19307203 TI - Question 1. Does melatonin improve sleep pattern in children with attention deficit hyperactivity disorder? PMID- 19307204 TI - Towards evidence based medicine for paediatricians. PMID- 19307205 TI - Question 2. Should fragile X be tested for in boys with a diagnosis of autistic spectrum disorder? PMID- 19307206 TI - Question 3. Is continuous positive airway pressure effective in bronchiolitis? PMID- 19307208 TI - Antibodies to reticulocyte binding protein-like homologue 4 inhibit invasion of Plasmodium falciparum into human erythrocytes. AB - Plasmodium falciparum invasion into human erythrocytes relies on the interaction between multiple parasite ligands and their respective erythrocyte receptors. The sialic acid-independent invasion pathway is dependent on the expression of P. falciparum reticulocyte binding protein-like homologue 4 (PfRh4), as disruption of the gene abolishes the ability of parasites to switch to this pathway. We show that PfRh4 is present as an invasion ligand in culture supernatants as a 160-kDa proteolytic fragment. We confirm that PfRh4 binds to the surfaces of erythrocytes through recognition of an erythrocyte receptor that is neuraminidase resistant but trypsin and chymotrypsin sensitive. Serum antibodies from malaria-exposed individuals show reactivity against the binding domain of PfRh4. Purified immunoglobulin G raised in rabbits against the binding domain of PfRh4 blocked the binding of native PfRh4 to the surfaces of erythrocytes and inhibited erythrocyte invasion of parasites using sialic acid-independent invasion pathways and grown in neuraminidase-treated erythrocytes. Our results suggest PfRh4 is a potential vaccine candidate. PMID- 19307209 TI - Cryptococcus neoformans directly stimulates perforin production and rearms NK cells for enhanced anticryptococcal microbicidal activity. AB - NK cells, in addition to possessing antitumor and antiviral activity, exhibit perforin-dependent microbicidal activity against the opportunistic pathogen Cryptococcus neoformans. However, the factors controlling this response, particularly whether the pathogen itself provides an activation or rearming signal, are largely unknown. The current studies were performed to determine whether exposure to this fungus alters subsequent NK cell anticryptococcal activity. NK cells lost perforin and mobilized lysosome-associated membrane protein 1 to the cell surface following incubation with the fungus, indicating that degranulation had occurred. Despite a reduced perforin content during killing, NK cells acquired an enhanced ability to kill C. neoformans, as demonstrated using auxotrophs that allowed independent assessment of the killing of two strains. De novo protein synthesis was required for optimal killing; however, there was no evidence that a soluble factor contributed to the enhanced anticryptococcal activity. Exposure of NK cells to C. neoformans caused the cells to rearm, as demonstrated by increased perforin mRNA levels and enhanced loss of perforin when transcription was blocked. Degranulation alone was insufficient to provide the activation signal as NK cells lost anticryptococcal activity following treatment with strontium chloride. However, NK cells regained the activity upon prolonged exposure to C. neoformans, which is consistent with activation by the microbe. The enhanced cytotoxicity did not extend to tumor killing since NK cells exposed to C. neoformans failed to kill NK-sensitive tumor targets (K562 cells). These studies demonstrate that there is contact-mediated microbe-specific rearming and activation of microbicidal activity that are necessary for optimal killing of C. neoformans. PMID- 19307210 TI - Campylobacter jejuni glycosylation island important in cell charge, legionaminic acid biosynthesis, and colonization of chickens. AB - Previously, we identified five genes (Cj1321 to Cj1326, of which Cj1325 and Cj1326 are a single gene) in the O-linked flagellin glycosylation island that are highly prevalent in Campylobacter jejuni isolates from chickens. We report mutagenesis, functional, and structural data to confirm that this locus, and Cj1324 in particular, has a significant contributory role in the colonization of chickens by C. jejuni. A motile DeltaCj1324 mutant with intact flagella was considerably less hydrophobic and less able to autoagglutinate and form biofilms than the parent strain, 11168H, suggesting that the surface charge of flagella of Cj1324-deficient strains was altered. The physical and functional attributes of the parent were restored upon complementation. Structural analysis of flagellin protein purified from the DeltaCj1324 mutant revealed the absence of two legionaminic acid glycan modifications that were present in the parent strain, 11168H. These glycoform modifications were shown to be prevalent in chicken isolates and confirm that differences in the highly variable flagellin glycosylation locus can relate to the strain source. The discovery of molecular mechanisms influencing the persistence of C. jejuni in poultry aids the rational design of approaches to control this problematic pathogen in the food chain. PMID- 19307211 TI - The plasmid of Escherichia coli strain S88 (O45:K1:H7) that causes neonatal meningitis is closely related to avian pathogenic E. coli plasmids and is associated with high-level bacteremia in a neonatal rat meningitis model. AB - A new Escherichia coli virulent clonal group, O45:K1, belonging to the highly virulent subgroup B2(1) was recently identified in France, where it accounts for one-third of E. coli neonatal meningitis cases. Here we describe the sequence, epidemiology and function of the large plasmid harbored by strain S88, which is representative of the O45:K1 clonal group. Plasmid pS88 is 133,853 bp long and contains 144 protein-coding genes. It harbors three different iron uptake systems (aerobactin, salmochelin, and the sitABCD genes) and other putative virulence genes (iss, etsABC, ompT(P), and hlyF). The pS88 sequence is composed of several gene blocks homologous to avian pathogenic E. coli plasmids pAPEC-O2-ColV and pAPEC-O1-ColBM. PCR amplification of 11 open reading frames scattered throughout the plasmid was used to investigate the distribution of pS88 and showed that a pS88-like plasmid is present in other meningitis clonal groups such as O18:K1, O1:K1, and O83:K1. A pS88-like plasmid was also found in avian pathogenic strains and human urosepsis strains belonging to subgroup B2(1). A variant of S88 cured of its plasmid displayed a marked loss of virulence relative to the wild-type strain in a neonatal rat model, with bacteremia more than 2 log CFU/ml lower. The salmochelin siderophore, a known meningovirulence factor, could not alone explain the plasmid's contribution to virulence, as a salmochelin mutant displayed only a minor fall in bacteremia (0.9 log CFU/ml). Thus, pS88 is a major virulence determinant related to avian pathogenic plasmids that has spread not only through meningitis clonal groups but also human urosepsis and avian pathogenic strains. PMID- 19307212 TI - Cytolethal distending toxin promotes Helicobacter cinaedi-associated typhlocolitis in interleukin-10-deficient mice. AB - Helicobacter cinaedi colonizes a wide host range, including rodents, and may be an emerging zoonotic agent. Colonization parameters, pathology, serology, and inflammatory responses to wild-type H. cinaedi (WT(Hc)) were evaluated in B6.129P2-IL-10(tm1Cgn) (IL-10(-/-)) mice for 36 weeks postinfection (WPI) and in C57BL/6 (B6) mice for 12 WPI. Because cytolethal distending toxin (CDT) may be a virulence factor, IL-10(-/-) mice were also infected with the cdtB(Hc) and cdtB N(Hc) isogenic mutants and evaluated for 12 WPI. Consistent with other murine enterohepatic helicobacters, WT(Hc) did not cause typhlocolitis in B6 mice, but mild to severe lesions developed at the cecocolic junction in IL-10(-/-) mice, despite similar colonization levels of WT(Hc) in the cecum and colon of both B6 and IL-10(-/-) mice. WT(Hc) and cdtB mutants also colonized IL-10(-/-) mice to a similar extent, but infection with either cdtB mutant resulted in attenuated typhlocolitis and hyperplasia compared to infection with WT(Hc) (P < 0.03), and only WT(Hc) infection caused dysplasia and intramucosal carcinoma. WT(Hc) and cdtB(Hc) mutant infection of IL-10(-/-) mice elevated mRNA expression of tumor necrosis factor alpha, inducible nitric oxide synthase, and gamma interferon in the cecum, as well as elevated Th1-associated serum immunoglobulin G2a(b) compared to infection of B6 mice (P < 0.05). Although no hepatitis was noted, liver samples were PCR positive at various time points for WT(Hc) or the cdtB(Hc) mutant in approximately 33% of IL-10(-/-) mice and in 10 to 20% of WT(Hc) infected B6 mice. These results indicate that WT(Hc) can be used to model inflammatory bowel disease in IL-10(-/-) mice and that CDT contributes to the virulence of H. cinaedi. PMID- 19307213 TI - Induction of adhesion-inhibitory antibodies against placental Plasmodium falciparum parasites by using single domains of VAR2CSA. AB - In areas of endemicity pregnancy-associated malaria is an important cause of maternal anemia, stillbirth, and delivery of low-birth-weight children. The syndrome is precipitated by the accumulation of Plasmodium falciparum-infected erythrocytes in the placenta, mediated through an interaction between a parasite protein expressed on erythrocytes named variant surface antigen 2-chondroitin sulfate A (VAR2CSA) and CSA on syncytiotrophoblasts. VAR2CSA is a large polymorphic protein consisting of six Duffy binding-like (DBL), domains and with current constraints on recombinant protein production it is not possible to produce entire VAR2CSA recombinant proteins. Furthermore, the presence of polymorphisms has raised the question of whether it is feasible to define VAR2CSA antigens eliciting broadly protective antibodies. Thus, the challenge for vaccine development is to define smaller parts of the molecule which induce antibodies that inhibit CSA binding of different parasite strains. In this study, we produced a large panel of VAR2CSA proteins and raised antibodies against these antigens. We show that antibodies against the DBL4 domain effectively inhibit parasite binding. As the inhibition was not limited to homologous parasite strains, it seems feasible to base a protective malaria vaccine on a single VAR2CSA DBL domain. PMID- 19307214 TI - Silencing of host cell CYBB gene expression by the nuclear effector AnkA of the intracellular pathogen Anaplasma phagocytophilum. AB - Coevolution of intracellular bacterial pathogens and their host cells resulted in the appearance of effector molecules that when translocated into the host cell modulate its function, facilitating bacterial survival within the hostile host environment. Some of these effectors interact with host chromatin and other nuclear components. In this report, we show that the AnkA protein of Anaplasma phagocytophilum, which is translocated into the host cell nucleus, interacts with gene regulatory regions of host chromatin and is involved in downregulating expression of CYBB (gp91(phox)) and other key host defense genes. AnkA effector protein rapidly accumulated in nuclei of infected cells coincident with changes in CYBB transcription. AnkA interacted with transcriptional regulatory regions of the CYBB locus at sites where transcriptional regulators bind. AnkA binding to DNA occurred at regions with high AT contents. Mutation of AT stretches at these sites abrogated AnkA binding. Histone H3 acetylation decreased dramatically at the CYBB locus during A. phagocytophilum infection, particularly around AnkA binding sites. Transcription of CYBB and other defense genes was significantly decreased in AnkA-transfected HL-60 cells. These data suggest a mechanism by which intracellular pathogens directly regulate host cell gene expression mediated by nuclear effectors and changes in host chromatin structure. PMID- 19307215 TI - Diversity of Ehrlichia ruminantium major antigenic protein 1-2 in field isolates and infected sheep. AB - Proteins expressed from the map1 multigene family of Ehrlichia ruminantium are strongly recognized by immune T and B cells from infected animals or from animals that were infected and have recovered from heartwater disease (although still remaining infected carriers). Analogous multigene clusters also encode the immunodominant outer membrane proteins (OMPs) in other ehrlichial species. Recombinant protein analogs of the expressed genes and DNA vaccines based on the multigene clusters have been shown to induce protective immunity, although this was less effective in heterologous challenge situations, where the challenge strain major antigenic protein 1 (MAP1) sequence differed from the vaccine strain MAP1. Recent data for several ehrlichial species show differential expression of the OMPs in mammalian versus tick cell cultures and dominant expression of individual family members in each type of culture system. However, many genes in the clusters appear to be complete and functional and to generate mRNA transcripts. Recent data also suggest that there may be a low level of protein expression from many members of the multigene family, despite primary high-level expression from an individual member. A continuing puzzle, therefore, is the biological roles of the different members of these OMP multigene families. Complete genome sequences are now available for two geographically divergent strains of E. ruminantium (Caribbean and South Africa strains). Comparison of these sequences revealed amino acid sequence diversity in MAP1 (89% identity), which is known to confer protection in a mouse model and to be the multigene family member primarily expressed in mammalian cells. Surprisingly, however, the greatest sequence diversity (79% identity) was in the less-studied map1-2 gene. We investigated here whether this map1-2 diversity was a general feature of E. ruminantium in different cultured African strains and in organisms from infected sheep. Comparison of MAP1-2s revealed amino acid identities of 75 to 100% (mean of 86%), compared to 84 to 100% (mean of 89%) for MAP1s. Interestingly, MAP1-2s varied independently of MAP1s such that E. ruminantium strains with similar MAP1s had diverse MAP1-2s and vice versa. Different MAP1-2s were found in individual infected sheep. Different regions of a protein may be subjected to different evolutionary forces because of recombination and/or selection, which results in those regions not agreeing with a phylogeny deduced from the whole molecule. This appears to be true for both MAP1 and MAP1-2, where statistical likelihood methods detect heterogeneous evolutionary rates for segments of both molecules. Sera from infected cattle recognized a MAP1-2 variable-region peptide in enzyme-linked immunosorbent assay, but less strongly and consistently than a MAP1 peptide (MAP1B). Heterologous protective immunity may depend on recognition of a complex set of varying OMP epitopes. PMID- 19307216 TI - Bacillus anthracis edema toxin suppresses human macrophage phagocytosis and cytoskeletal remodeling via the protein kinase A and exchange protein activated by cyclic AMP pathways. AB - Bacillus anthracis, the etiological agent of anthrax, is a gram-positive spore forming bacterium. It produces edema toxin (EdTx), a powerful adenylate cyclase that increases cyclic AMP (cAMP) levels in host cells. Because other cAMP increasing agents inhibit key macrophage (MPhi) functions, such as phagocytosis, it was hypothesized that EdTx would exhibit similar suppressive activities. Our previous GeneChip data showed that EdTx downregulated MPhi genes involved in actin cytoskeleton remodeling, including protein kinase A (PKA). To further examine the role of EdTx during anthrax pathogenesis, we explored the hypothesis that EdTx treatment leads to deregulation of the cAMP-dependent PKA system, resulting in impaired cytoskeletal functions essential for MPhi activity. Our data revealed that EdTx significantly suppressed human MPhi phagocytosis of Ames spores. Cytoskeletal changes, such as decreased cell spreading and lowered F actin content, were also observed for toxin-treated MPhis. Further, EdTx altered the protein levels and activity of PKA and exchange protein activated by cAMP (Epac), a recently identified cAMP-binding molecule. By using PKA- and Epac selective cAMP analogs, we confirmed the involvement of both pathways in the inhibition of MPhi functions elicited by EdTx-generated cAMP. These results suggested that EdTx weakened the host immune response by increasing cAMP levels, which then signaled via PKA and Epac to cripple MPhi phagocytosis and interfered with cytoskeletal remodeling. PMID- 19307217 TI - Reproducible community dynamics of the gastrointestinal microbiota following antibiotic perturbation. AB - Shifts in microbial communities are implicated in the pathogenesis of a number of gastrointestinal diseases, but we have limited understanding of the mechanisms that lead to altered community structures. One difficulty with studying these mechanisms in human subjects is the inherent baseline variability of the microbiota in different individuals. In an effort to overcome this baseline variability, we employed a mouse model to control the host genotype, diet, and other possible influences on the microbiota. This allowed us to determine whether the indigenous microbiota in such mice had a stable baseline community structure and whether this community exhibited a consistent response following antibiotic administration. We employed a tag-sequencing strategy targeting the V6 hypervariable region of the bacterial small-subunit (16S) rRNA combined with massively parallel sequencing to determine the community structure of the gut microbiota. Inbred mice in a controlled environment harbored a reproducible baseline community that was significantly impacted by antibiotic administration. The ability of the gut microbial community to recover to baseline following the cessation of antibiotic administration differed according to the antibiotic regimen administered. Severe antibiotic pressure resulted in reproducible, long lasting alterations in the gut microbial community, including a decrease in overall diversity. The finding of stereotypic responses of the indigenous microbiota to ecologic stress suggests that a better understanding of the factors that govern community structure could lead to strategies for the intentional manipulation of this ecosystem so as to preserve or restore a healthy microbiota. PMID- 19307218 TI - Iron-regulated surface determinant protein A mediates adhesion of Staphylococcus aureus to human corneocyte envelope proteins. AB - The ability of Staphylococcus aureus to colonize the human nares is a crucial prerequisite for disease. IsdA is a major S. aureus surface protein that is expressed during human infection and required for nasal colonization and survival on human skin. In this work, we show that IsdA binds to involucrin, loricrin, and cytokeratin K10, proteins that are present in the cornified envelope of human desquamated epithelial cells. To measure the forces and dynamics of the interaction between IsdA and loricrin (the most abundant protein of the cornified envelope), single-molecule force spectroscopy was used, demonstrating high specificity binding. IsdA acts as a cellular adhesin to the human ligands, promoting whole-cell binding to immobilized proteins, even in the absence of other S. aureus components (as shown by heterologous expression in Lactococcus lactis). Inhibition experiments revealed the binding of the human ligands to the same IsdA region. This region was mapped to the NEAT domain of IsdA. The NEAT domain also was found to be required for S. aureus whole-cell binding to the ligands as well as to human nasal cells. Thus, IsdA is an important adhesin to human ligands, which predominate in its primary ecological niche. PMID- 19307219 TI - Resistance of Capnocytophaga canimorsus to killing by human complement and polymorphonuclear leukocytes. AB - Capnocytophaga canimorsus is a bacterium of the canine oral flora known since 1976 to cause rare but severe septicemia and peripheral gangrene in patients that have been in contact with a dog. It was recently shown that these bacteria do not elicit an inflammatory response (H. Shin, M. Mally, M. Kuhn, C. Paroz, and G. R. Cornelis, J. Infect. Dis. 195:375-386, 2007). Here, we analyze their sensitivity to the innate immune system. Bacteria from the archetype strain Cc5 were highly resistant to killing by complement. There was little membrane attack complex (MAC) deposition in spite of C3b deposition. Cc5 bacteria were as resistant to phagocytosis by human polymorphonuclear leukocytes (PMNs) as Yersinia enterocolitica MRS40, endowed with an antiphagocytic type III secretion system. We isolated Y1C12, a transposon mutant that is hypersensitive to killing by complement via the antibody-dependent classical pathway. The mutation inactivated a putative glycosyltransferase gene, suggesting that the Y1C12 mutant was affected at the level of a capsular polysaccharide or lipopolysaccharide (LPS) structure. Cc5 appeared to have several polysaccharidic structures, one being altered in Y1C12. The structure missing in Y1C12 could be purified by classical LPS purification procedures and labeled by tritiated palmitate, indicating that it is more likely to be an LPS structure than a capsule. Y1C12 bacteria were also more sensitive to phagocytosis by PMNs than wild-type bacteria. In conclusion, a polysaccharide structure, likely an LPS, protects C. canimorsus from deposition of the complement MAC and from efficient phagocytosis by PMNs. PMID- 19307221 TI - Visual assessment of jaundice in term and late preterm infants. AB - OBJECTIVE: To determine the accuracy of predischarge visual assessment of jaundice for estimating bilirubin concentration and predicting risk of significant neonatal hyperbilirubinaemia. DESIGN: Prospective cohort study. SETTING: Well Baby Nursery at the Hospital of the University of Pennsylvania. PATIENTS: 522 term and late preterm newborns. INTERVENTIONS: Nurses used a 5 point scale to grade the maximum cephalocaudal extent of jaundice prior to discharge. MAIN OUTCOME MEASURES: (1) Correlation between jaundice grade and bilirubin concentration. (2) Predictive accuracy of jaundice grade for identifying infants who developed significant hyperbilirubinaemia, defined as a bilirubin level that at any time after birth exceeded or was within 1 mg/dl (17 micromol/l) of the American Academy of Pediatrics-recommended hour-specific phototherapy treatment threshold. RESULTS: Nurses' assessment of jaundice extent was only moderately correlated with bilirubin concentration and was similar in black and non-black infants (Spearman's rho = 0.45 and 0.55, respectively (p = 0.13)). The correlation was particularly weak among infants <38 weeks' gestational age (rho = 0.29) compared with infants > or = 38 weeks' gestation (rho = 0.53, p = 0.05). Jaundice extent had poor overall accuracy for predicting risk of significant hyperbilirubinaemia (c-statistic = 0.65) but complete absence of jaundice had high sensitivity (95%) and excellent negative predictive value (99%) for ruling out the development of significant hyperbilirubinaemia. CONCLUSIONS: Clinicians should not use extent of cephalocaudal jaundice progression to estimate bilirubin levels during the birth hospitalisation, especially in late preterm infants. However, the complete absence of jaundice can be used to predict with very high accuracy which infants will not develop significant hyperbilirubinaemia. PMID- 19307220 TI - Antibody-enhanced, Fc gamma receptor-mediated endocytosis of Clostridium difficile toxin A. AB - Toxin A (TcdA) and toxin B (TcdB) are major virulence factors of Clostridium difficile. These two toxins intoxicate cultured cells by similar mechanisms, and TcdB generally is more potent than TcdA in cultured cells. The exact reason for this difference is unclear. Here, we report that the cellular effects of TcdA can be substantially enhanced via an opsonizing antibody through Fc gamma receptor I (FcgammaRI)-mediated endocytosis. A TcdA-specific monoclonal antibody, A1H3, was found to significantly enhance the cytotoxicity of TcdA to macrophages and monocytes. The A1H3-dependent enhancement of glucosyltransferase activity, cytoskeleton disruption, and tumor necrosis factor alpha production induced by TcdA was further demonstrated using RAW 264.7 cells. Subsequent experiments indicated that the interaction of FcgammaRI with A1H3 underlays the antibody dependent enhancement of the cellular effects of TcdA. While blocking FcgammaRII and FcgammaRIII with anti-CD16/32 antibodies did not affect the TcdA-mediated glucosylation of Rac1 in RAW 264.7 cells, presaturation of FcgammaRI with anti CD64 antibodies in THP1 cells significantly reduced this activity. Incubation of a TcdA-A1H3 immune complex with recombinant mouse CD64 completely abrogated the A1H3-mediated enhancement of the glucosyltransferase activity of TcdA in RAW 264.7 cells. Moreover, expression of FcgammaRI in CHO cells strikingly enhanced the sensitivity of these cells to TcdA complexed with A1H3. We showed that the presence of A1H3 facilitated cell surface recruitment of TcdA, contributing to the antibody-dependent, FcgammaRI-mediated enhancement of TcdA activity. Finally, studies using chlorpromazine and endosomal acidification inhibitors revealed an important role of the endocytic pathway in the A1H3-dependent enhancement of TcdA activity. PMID- 19307222 TI - Atropine, fentanyl and succinylcholine for non-urgent intubations in newborns. AB - OBJECTIVE: Describe intubation conditions and adverse events when using atropine fentanyl +/- succinylcholine as premedication. DESIGN: Prospective observational study, as part of a quality improvement initiative. SETTING: Two level 3 neonatal intensive care units in Ottawa, Canada PATIENTS: 60 infants, median 27 weeks gestation, 1023 g at birth were included. INTERVENTIONS: Infants received atropine, fentanyl +/- succinylcholine prior to the intubation. Succinylcholine was given for all infants > or =34 weeks and at the discretion of the physician for those <34 weeks. MAIN OUTCOME MEASURES: The primary outcome was the number of attempts. Secondary outcomes were number of attempts and intubation conditions in infants who received and those who did not receive succinylcholine and safety. RESULTS: The median number of attempts was 2. 91.7% had excellent or good conditions. The median number of attempts for infants who received succinylcholine was lower (1 vs 2) than those who did not. No serious adverse events were noted. 2 cases of difficult bag and mask ventilation after administration of fentanyl were noted. CONCLUSION: Atropine, fentanyl and succinylcholine before non-urgent intubations in newborns has led to a low number of attempts and good intubation conditions with no adverse events. PMID- 19307223 TI - Very preterm children show impairments across multiple neurodevelopmental domains by age 4 years. AB - OBJECTIVES: Neurodevelopmental outcomes associated with preterm birth are of major health and educational concern. This study examined the neuromotor, cognitive, language and emotional/behavioural outcomes of a regional cohort of 4 year-old children born extremely preterm (EPT: 23-27 weeks' gestation), very preterm (VPT: 28-33 weeks) and full term (FT: 38-41 weeks). Of particular interest were children's risks of impairment across multiple neurodevelopmental domains. METHODS: Data were gathered as part of a prospective longitudinal study of 105 very preterm (< or = 33 weeks gestation) and 107 FT children born during 1998-2000. At 4 years corrected age, children underwent a comprehensive multidisciplinary assessment that included a paediatric neurological examination, cognitive and language testing, and an assessment of child emotional and behavioural adjustment. RESULTS: At age 4 years, compared to FT children, EPT and VPT children had increased risks of cerebral palsy (EPT 18%, VPT 15%, FT 1%), cognitive delay (EPT 33%, VPT 36%, FT 13%), language delay (EPT 29%, VPT 29%, FT 10%) and emotional/behavioural adjustment problems (EPT 37%, VPT 13%, FT 11%). EPT and VPT children were three times more likely to have multiple domain impairments than FT children (EPT 30%, VPT 29%, FT 10%). CONCLUSIONS: A substantial proportion of preschool children born very preterm show clinically significant problems in at least one neurodevelopmental domain, with impairment in multiple domains being common. There is a need to monitor preschool development across a range of functional domains and to consider the likely cascading effects of multiple impairments on later development. PMID- 19307224 TI - Cardiac troponin I at birth is of fetal-neonatal origin. AB - OBJECTIVE: Neonates produce predominantly skeletal muscle troponin I (TnI) in the myocardium; however, in asphyxiated neonates, high levels of cardiac troponin I (cTnI) have been found. We hypothesised that in these circumstances cTnI could be from the mother or the result of a change in fetal/neonatal production in response to an insult. In this study, we aimed to compare cTnI concentrations in asphyxiated neonates with those of their respective mothers. METHODS: In this prospective observational study, we enrolled all asphyxiated neonates transferred by the Veneto Region Neonatal Transport Service in the period 1 January 2006 to 31 March 2007. Asphyxia was defined as a pH < or =7.00 and/or a base deficit of > or =16 mmol per litre. Neonatal and maternal blood samples were obtained for cTnI determination. RESULTS: We enrolled 19 asphyxiated neonates (median gestational age: 39 weeks, interquartile range 34-40; birth weight 3100 g, 1950-3340). Their cTnI concentrations were significantly higher in comparison with their mothers: 0.24 microg/l (0.13-0.50) vs 0.04 microg/l (0.04-0.04); p<0.01. CONCLUSIONS: Increased cTnI concentrations detected in asphyxiated neonates are of neonatal origin and are not derived from the mother. In asphyxiated neonates, there may be predisposing factors that could cause earlier switching from skeletal TnI to cTnI in the myocardium. PMID- 19307225 TI - Gastrooesophageal reflux disease in preterm infants: current management and diagnostic dilemmas. AB - Gastrooesophageal reflux disease (GORD) provides a diagnostic and therapeutic challenge to many neonatologists. Reflux of gastric contents is common in preterm infants but usually not pathological. GORD is frequently diagnosed despite the lack of a fully identified clinical syndrome and of a truly valid diagnostic test. Treatment modalities, for which there is little convincing evidence regarding efficacy, are commonly instigated for troublesome symptoms attributed to GORD. Diagnosis is so problematic in preterm infants that GORD is starting to be described as the clinical syndrome that responds to anti-reflux treatment. We discuss the dilemmas facing us when dealing with this condition, summarise the best available evidence regarding diagnosis and management, and use it to inform a suggested treatment pathway. We introduce the concept of a clinical scoring system to aid the diagnosis and monitoring of GORD in preterm infants and highlight areas where further research would be beneficial. PMID- 19307226 TI - Ultrasound structural fetal anomaly screening: an update. AB - Screening for fetal anomalies by an ultrasound scan was introduced in many maternity units in the UK in the late 1970s and has become routine in most since then although there is little uniformity as to how the scan is performed and when it is offered. Up to 5% of babies are born with a congenital abnormality. In this review article we will focus on what is already known about screening for the commonest and most serious structural fetal anomalies at the 11-14 and 18-23 week scans, and discuss new techniques that promise to improve the accuracy of screening and diagnosis. Chromosomal abnormalities, "soft markers" and biochemical screening are beyond the scope of this review. PMID- 19307228 TI - Effect of parental age at birth on the accumulation of deficits, frailty and survival in older adults. AB - INTRODUCTION: parental age at conception may affect life expectancy. Adult daughters of older fathers seem to live shorter lives and, in one study, being born to a mother aged <25 was an important predictor of exceptional longevity. The effect of parental age on fitness/frailty in late life is unknown. We aimed to investigate the relationships between parental age and frailty and longevity in older adults. METHODS: in the Canadian Study of Health and Aging (CSHA), data was collected on individuals aged >or=65 using a Self-Assessed Risk Factor Questionnaire and screening interview. In this secondary analysis, 5112 participants had complete data for parental age, frailty status and 10-year survival. Parental age was divided into three groups, with cut-offs at 25 and 45 for fathers and at 25 and 40 for mothers. Frailty was defined by an index of deficits. Survival was analysed using Kaplan-Meier curves and Cox regression with analyses adjusted for subject's age, sex and age of the other parent. RESULTS: mean maternal age at subject's birth was 29.2y (SD 6.8) and mean paternal age 33.3y (SD 7.8). There was no effect of maternal or paternal age on survival for either sons or daughters. Similarly, there was no association between parental age and subject frailty in old age. CONCLUSION: we did not identify an association between parental age and frailty or longevity in older adult participants in the CSHA. PMID- 19307229 TI - Associations of resting heart rate with insulin resistance, cardiovascular events and mortality in chronic kidney disease. AB - BACKGROUND: Insulin resistance is associated with increased sympathetic and reduced parasympathetic activity. Resting heart rate reflects autonomic activity. Therefore, we examined the associations of resting heart rate with insulin resistance, cardiovascular events and mortality in the moderate chronic kidney disease (CKD) population. METHODS: Four hundred and sixty participants with MDRD GFR <60 ml/min/1.73 m(2) in the limited access Atherosclerosis Risk in Communities (ARIC) study database were divided into four resting heart rate groups: <60, 60-74, 75-89 and >or=90/min. The prevalence of metabolic syndrome at baseline across the groups was examined. Time to cardiovascular composite (myocardial infarction or fatal coronary artery disease event or stroke or coronary revascularization procedure) and time to all-cause death were examined in multivariate Cox models. RESULTS: The prevalence of metabolic syndrome in the <60, 60-74, 75-89 and >or=90/min groups were 41, 44, 69 and 82% (P < 0.001), respectively. In a multivariate Cox model adjusted for demographics, comorbidity, haemoglobin and physical activity, compared to the 60-74/min group, the hazard ratios of cardiovascular composite in <60, 75-89 and >or=90/min groups were 1.27 (95% CI 0.75-2.16), 1.79 (95% CI 1.07-2.99) and 1.37 (95% CI 0.54-3.44), respectively. In a similar model, the hazard ratios of death were 1.47 (95% CI 0.85-2.53), 3.11 (95% CI 1.93-5.02) and 3.97 (95% CI 1.99-7.94), respectively. CONCLUSIONS: Resting heart rate is associated with metabolic syndrome in moderate CKD. Higher resting heart is associated with increased mortality and possibly cardiovascular events in this population. Interventional studies to examine whether a target resting heart rate of 60-74/min improves cardiovascular outcomes and survival in moderate CKD are warranted. PMID- 19307230 TI - Alcohol consumption and 5-year onset of chronic kidney disease: the AusDiab study. AB - BACKGROUND: Excessive alcohol consumption is a risk factor for hypertension and stroke; however, evidence for an association with chronic kidney disease is conflicting. METHODS: A total of 6259 adults >or=25 years of age, without a history of alcohol dependence, participating in baseline (1999-2000) and follow up (2004-2005) phases of an Australian population-representative study (AusDiab) were the subject of this analysis. Alcohol consumption status and volume/frequency were collected by standardized interviewer administered questionnaires and self-administered food frequency questionnaires. The outcomes were as follows: (i) 5-year decline in estimated glomerular filtration rate (eGFR) >or=10%, with baseline eGFR >or= 60 and final eGFR <60 mL/min/1.73 m(2), and (ii) 5-year doubling of albumin to creatinine ratio (ACR) with final ACR >or= 2.5 (males)/>or= 3.5 (females) mg/mmol, in the absence of albuminuria at baseline. RESULTS: Self-identification as a moderate or heavy, versus light, drinker was associated with elevated risk of albuminuria in males and females <65 years of age (OR, 95% CI: males 1.87, 0.99-3.52; females 2.38, 1.37-4.14). Odds of de novo eGFR <60 mL/min/1.73 m(2) were 0.34 (95% CI 0.22-0.59) and 0.68 (95% CI 0.36-1.27) in males and females, respectively, who were moderate-heavy drinkers. Alcohol intake of >or=30 g/day was associated with an increased risk of albuminuria after adjustment for age, sex and baseline kidney function (OR = 1.59, 95% CI 1.07-2.36), but a reduced risk of eGFR <60 mL/min/1.73 m(2) (OR = 0.59, 95% CI 0.37-0.95), compared with consumption of <10 g/day. CONCLUSIONS: Moderate-heavy alcohol consumption may be an important modifiable risk factor for albuminuria in the general population. The natural history of alcohol-induced kidney damage and how this relates to markers of kidney function in the general population warrant further research. PMID- 19307231 TI - Valsartan regulates the interaction of angiotensin II type 1 receptor and endothelial nitric oxide synthase via Src/PI3K/Akt signalling. AB - AIMS: Valsartan, a selective angiotensin II type 1 receptor (AT1R) blocker, has beneficial effects in the cardiovascular system in part by its increase of nitric oxide (NO) bioavailability, yet the mechanisms are unclear. We investigated the molecular mechanisms underlying this effect in endothelial cells (ECs). METHODS AND RESULTS: NO production was examined by Griess reagent assay, DAF-2 DA fluorescence staining and cGMP ELISA kits. Protein interaction was determined by western blotting and immunoprecipitation. Treating bovine or human aortic ECs with valsartan increased NO production, as evidenced by elevated level of stable NO metabolites and intracellular cGMP. Valsartan increased the phosphorylation but not the protein level of endothelial NO synthase (eNOS). Inhibition of phosphoinositide-3 kinase (PI3K)/Akt and Src pathways by specific inhibitors suppressed valsartan-induced NO release. In addition, valsartan increased the tyrosine residue phosphorylation of AT1R, which was attenuated by inhibition of Src but not PI3K activities. Valsartan also suppressed the interaction of eNOS and AT1R, which was blocked by Src or PI3K inhibition. CONCLUSION: Valsartan induced NO production in ECs is mediated through Src/PI3K/Akt-dependent phosphorylation of eNOS. Valsartan-induced AT1R phosphorylation depends on Src but not PI3K, whereas valsartan-induced suppression of AT1R-eNOS interaction depends on Src/PI3K/Akt signalling. These results indicate a novel vasoprotective mechanism of valsartan in upregulating NO production in ECs. PMID- 19307232 TI - Antithrombin reduces shedding of the endothelial glycocalyx following ischaemia/reperfusion. AB - AIMS: Antithrombin is an important inhibitor of the coagulation system, additionally exerting specific anti-inflammatory effects on endothelial cells. Healthy vascular endothelium is coated by the endothelial glycocalyx, diminution of which increases capillary permeability, e.g. after ischaemia. Antithrombin is known to infiltrate the glycocalyx, binding to glycosaminoglycans, and to preserve the glycocalyx after application tumour necrosis factor-alpha. We investigated the influence of antithrombin on glycocalyx subjected to ischaemia/reperfusion. METHODS AND RESULTS: Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer (KHB). Antithrombin was applied to achieve physiological levels (1 U/mL) before inducing 20 min of ischaemia (37 degrees C). Hearts were reperfused for 20 min at constant flow (baseline perfusion pressure 70 cmH(2)O) with KHB or KHB plus 2 g% hydroxyethyl starch (130 kDa). Coronary net fluid filtration was assessed directly by measuring transudate formation on the epicardial surface. Post-ischaemic coronary release of syndecan-1 and heparan sulfate was quantified by ELISA. Hearts were perfusion-fixed to visualize the glycocalyx by electron microscopy. Ischaemia/reperfusion caused degradation of the glycocalyx, enhanced coronary perfusion pressure, and increased vascular permeability. Antithrombin significantly reduced post-ischaemic glycocalyx shedding, coronary perfusion pressure, coronary leak, and tissue oedema formation compared to untreated hearts. Additional application of colloid augmented these actions of antithrombin. Electron microscopy revealed a mostly intact glycocalyx after antithrombin treatment. CONCLUSION: Antithrombin preserves the endothelial glycocalyx, sustaining the vascular barrier function and reducing interstitial oedema. The potentiated effect of colloid in these hearts suggests that the prevention of shedding should be of functional benefit also in vivo. PMID- 19307233 TI - Assessment of radiation hazards due to natural radioactivity in some building materials used in Egyptian dwellings. AB - Different types of Egyptian building materials from various locations in Cairo and its suburbs have been analysed for natural radioactivity using gamma ray spectrometry. Concentrations of (226)Ra, (232)Th and (40)K were in the ranges of (12 +/- 2.8-65 +/- 6.5), (5 +/- 1.8-60 +/- 6.7) and (159 +/- 3.8-920 +/- 12.7 Bq kg(-1)), respectively. The minimum concentration of (226)Ra, (232)Th and (40)K was found in gravel samples, whereas the maximum in granite samples. The results are compared with the published data of other countries and with the world average limits. The radiological hazard parameters: radium equivalent activity, gamma index, alpha index, absorbed dose rate and the annual exposure rate, were determined to assess the radiation hazards associated with Egyptian buildings. All studied samples are lower than world average limits. PMID- 19307234 TI - Evaluation of transmission data of diagnostic X rays through concrete using Monte Carlo simulation. AB - The transmittance of diagnostic X-ray beams through concrete of two different densities were measured. Broad-beam X-ray transmissions were estimated using a Monte Carlo simulation at tube voltages of 50-150 kV and compared with National Council on Radiation Protection and Measurements (NCRP) data. The observed levels were higher than those predicted by the NCRP data; that is, the NCRP data underestimate the leakage radiation from widely used inverter systems. Some of the issues were resolved and new data were proposed. PMID- 19307235 TI - Neutron dose study with bubble detectors aboard the International Space Station as part of the Matroshka-R experiment. AB - As part of the Matroshka-R experiments, a spherical phantom and space bubble detectors (SBDs) were used on board the International Space Station to characterise the neutron radiation field. Seven experimental sessions with SBDs were carried out during expeditions ISS-13, ISS-14 and ISS-15. The detectors were positioned at various places throughout the Space Station, in order to determine dose variations with location and on/in the phantom in order to establish the relationship between the neutron dose measured externally to the body and the dose received internally. Experimental data on/in the phantom and at different locations are presented. PMID- 19307236 TI - Aromatic DNA adducts and polymorphisms in metabolic genes in healthy adults: findings from the EPIC-Spain cohort. AB - Aromatic compounds such as polycyclic aromatic hydrocarbons, arylamines and heterocyclic amines require metabolic activation to form metabolites able to bind to DNA, a process mediated by polymorphic enzymes. We measured aromatic DNA adducts in white blood cells by the (32)P-post-labelling assay in a sample of 296 healthy adults (147 men and 149 women) from five regions of Spain. We also analyzed functional polymorphisms in the metabolic genes CYP1A1, CYP1A2, EPHX1, GSTM1, GSTT1, NAT2 and SULT1A1. A significant increased level of DNA aromatic adducts was found related to the fast oxidation-hydrolysis phenotype defined by the polymorphism I462V in CYP1A1, the allele A in IVS1-154C>A of CYP1A2 and the combination Tyrosine-Arginine for Y113H and H139R of EPHX1. Geometric means (adducts per 10(-9) normal nucleotides) were 2.17, 4.04 and 6.30 for slow, normal and fast phenotypes, respectively (P-trend = 0.01). Slow acetylation by NAT2 was associated with a significant decrease in adduct level; subjects with slow alleles *5A and *7A/B had in average 1.56 x 10(-9)adducts, as compared with 5.60 for those with normal NAT2 activity (P-value = 0.01). No association was seen with polymorphisms of other metabolic genes such as GSTM1, GSTT1 or SULT1A1. We concluded that the metabolic pathways of oxidation, hydrolysis and acetylation are relevant to the formation of bulky DNA adducts. This could suggest a potential involvement of aromatic compounds in the formation of such adducts; however, given lack of specificity of the post-labeling assay, a firm conclusion cannot be drawn. PMID- 19307237 TI - Evolutionary Trace Annotation Server: automated enzyme function prediction in protein structures using 3D templates. AB - SUMMARY: The Evolutionary Trace Annotation (ETA) Server predicts enzymatic activity. ETA starts with a structure of unknown function, such as those from structural genomics, and with no prior knowledge of its mechanism uses the phylogenetic Evolutionary Trace (ET) method to extract key functional residues and propose a function-associated 3D motif, called a 3D template. ETA then searches previously annotated structures for geometric template matches that suggest molecular and thus functional mimicry. In order to maximize the predictive value of these matches, ETA next applies distinctive specificity filters -- evolutionary similarity, function plurality and match reciprocity. In large scale controls on enzymes, prediction coverage is 43% but the positive predictive value rises to 92%, thus minimizing false annotations. Users may modify any search parameter, including the template. ETA thus expands the ET suite for protein structure annotation, and can contribute to the annotation efforts of metaservers. AVAILABILITY: The ETA Server is a web application available at (http://mammoth.bcm.tmc.edu/eta/). PMID- 19307238 TI - Statistical model for whole genome sequencing and its application to minimally invasive diagnosis of fetal genetic disease. AB - There is currently great interest in the development of methods for the minimally invasive diagnosis of fetal genetic disease using cell-free DNA from maternal plasma samples obtained in the first trimester of pregnancy. With the rapid development of high-throughput sequencing technology, the possibility of detecting the presence of trisomy fetal genomes in the maternal plasma DNA sample has recently been explored. The major concern of this whole genome sequencing approach is that, while detecting the karyotype of the fetal genome from the maternal plasma requires extremely high accuracy of copy number estimation, the majority of the available high-throughput sequencing technologies require polymerase chain reaction (PCR) and are subject to the substantial bias that is inherent to the PCR process. We introduce a novel and sophisticated statistical model for the whole genome sequencing data, and based on this model, develop a highly sensitive method of Minimally Invasive Karyotyping (MINK) for the diagnosis of the fetal genetic disease. Specifically we demonstrate, by applying our statistical method to ultra high-throughput whole sequencing data, that trisomy 21 can be detected in a minor ('fetal') genome when it is mixed into a major ('maternal') background genome at frequencies as low as 5%. This observation provides additional proof of concept and justification for the further development of this method towards its eventual clinical application. Here, we describe the statistical and experimental methods that illustrate this approach and discuss future directions for technical development and potential clinical applications. PMID- 19307239 TI - KEGGgraph: a graph approach to KEGG PATHWAY in R and bioconductor. AB - MOTIVATION: KEGG PATHWAY is a service of Kyoto Encyclopedia of Genes and Genomes (KEGG), constructing manually curated pathway maps that represent current knowledge on biological networks in graph models. While valuable graph tools have been implemented in R/Bioconductor, to our knowledge there is currently no software package to parse and analyze KEGG pathways with graph theory. RESULTS: We introduce the software package KEGGgraph in R and Bioconductor, an interface between KEGG pathways and graph models as well as a collection of tools for these graphs. Superior to existing approaches, KEGGgraph captures the pathway topology and allows further analysis or dissection of pathway graphs. We demonstrate the use of the package by the case study of analyzing human pancreatic cancer pathway. AVAILABILITY: KEGGgraph is freely available at the Bioconductor web site (http://www.bioconductor.org). KGML files can be downloaded from KEGG FTP site (ftp://ftp.genome.jp/pub/kegg/xml). PMID- 19307240 TI - SBML2L(A)T(E)X: conversion of SBML files into human-readable reports. AB - SUMMARY: The XML-based Systems Biology Markup Language (SBML) has emerged as a standard for storage, communication and interchange of models in systems biology. As a machine-readable format XML is difficult for humans to read and understand. Many tools are available that visualize the reaction pathways stored in SBML files, but many components, e.g. unit declarations, complex kinetic equations or links to MIRIAM resources, are often not made visible in these diagrams. For a broader understanding of the models, support in scientific writing and error detection, a human-readable report of the complete model is needed. We present SBML2L(A)T(E)X, a Java-based stand-alone program to fill this gap. A convenient web service allows users to directly convert SBML to various formats, including DVI, L(A)T(E)X and PDF, and provides many settings for customization. AVAILABILITY: Source code, documentation and a web service are freely available at (http://www.ra.cs.uni-tuebingen.de/software/SBML2LaTeX). PMID- 19307241 TI - BioconductorBuntu: a Linux distribution that implements a web-based DNA microarray analysis server. AB - SUMMARY: BioconductorBuntu is a custom distribution of Ubuntu Linux that automatically installs a server-side microarray processing environment, providing a user-friendly web-based GUI to many of the tools developed by the Bioconductor Project, accessible locally or across a network. System installation is via booting off a CD image or by using a Debian package provided to upgrade an existing Ubuntu installation. In its current version, several microarray analysis pipelines are supported including oligonucleotide, dual-or single-dye experiments, including post-processing with Gene Set Enrichment Analysis. BioconductorBuntu is designed to be extensible, by server-side integration of further relevant Bioconductor modules as required, facilitated by its straightforward underlying Python-based infrastructure. BioconductorBuntu offers an ideal environment for the development of processing procedures to facilitate the analysis of next-generation sequencing datasets. AVAILABILITY: BioconductorBuntu is available for download under a creative commons license along with additional documentation and a tutorial from (http://bioinf.nuigalway.ie). PMID- 19307242 TI - Infernal 1.0: inference of RNA alignments. AB - SUMMARY: INFERNAL builds consensus RNA secondary structure profiles called covariance models (CMs), and uses them to search nucleic acid sequence databases for homologous RNAs, or to create new sequence- and structure-based multiple sequence alignments. AVAILABILITY: Source code, documentation and benchmark downloadable from http://infernal.janelia.org. INFERNAL is freely licensed under the GNU GPLv3 and should be portable to any POSIX-compliant operating system, including Linux and Mac OS/X. PMID- 19307243 TI - Percutaneous coronary intervention versus coronary artery bypass surgery in multivessel disease: a current perspective. AB - Coronary artery bypass surgery (CABG) and percutaneous coronary intervention (PCI) are both safe and established treatment modalities of invasive revascularization for patients with coronary artery disease (CAD). However, conflicting information exists when comparing the long-term efficacy of the two methods. The optimal treatment for patients with multivessel coronary artery disease (MVD) is still subject to discussion, given the lack of fairly designed, prospective, randomized data reflecting current practice in the modern era. Furthermore, the clinical outcomes after invasive revascularization differ according to the number of diseased vessels, presence or absence of diabetes, left main disease and left ventricular dysfunction. Hence, the question arises whether we should continue to use the term 'multivessel disease'. Conflicts of available data need to be addressed and overcome so that care of patients with CAD can be successfully tailored. In this review article we try to address some of the above conflicts, in an effort to improve our understanding in the care of patients with multivessel disease. We also provide an evidence-based perspective which may differ from the current standard of practice. PMID- 19307244 TI - Social vulnerability and unmet preventive care needs in outpatients of two French public hospitals. AB - BACKGROUND: Outpatients attending consultations at public hospitals may have unmet needs for preventive medical care. This study aimed to identify and assess the association between these needs, social vulnerability and mode of healthcare use. METHODS: In a multicentre epidemiological study, a group of socially vulnerable outpatients, was compared with a non-vulnerable group in a sample of 1316 outpatients selected in hospital consultations, using a validated tool for detection of social vulnerability. Before the patient was seen by medical staff, investigators collected data on social characteristics, healthcare use and preventive medical care received (interventions, advice). RESULTS: More than 75% of outpatients stated that they were regularly followed by a physician, usually a general practitioner, but fewer vulnerable than non-vulnerable outpatients were followed (77% vs. 89%, P < 10(-3)). For the majority of preventive interventions (vaccinations, screening for cardiovascular risk factors, gynaecological cancers), vulnerable outpatients presented a more marked shortage than non vulnerable patients, but there was an overall shortage in both groups. When recommended preventive interventions had not been delivered, they had rarely been offered in either group. After adjustment for mode of healthcare use, the differences in preventive care received persisted to the disadvantage of vulnerable outpatients with regard to technical preventive interventions, but there was no difference between the two groups regarding advice received to reduce risk behaviours. CONCLUSION: Unmet needs for preventive care primarily resulted from social inequalities in secondary access to such care. It may be necessary to set up specific interventions targeting vulnerable patients within hospital consultations. PMID- 19307245 TI - Inequalities in dental caries of 5-year-old children in Scotland, 1993-2003. AB - BACKGROUND: Previous research suggests there are significant differences between socio-economic groups in prevalence and amount of decayed missing and filled primary teeth (d3mft). The aim of this study was to describe the variation in obvious tooth decay experience amongst 5-year olds in Scotland and to look at the association between d3mft and deprivation in Scotland. METHODS: Data derived from 1993 to 2003 National Dental Inspection Programme were modelled using Bayesian multilevel zero-inflated Negative Binomial models, adjusting for age, sex and the deprivation. RESULTS: Deprivation is positively and significantly associated with having d3mft; the odds of a child in DepCat 7 (most deprived) having d3mft in 1993 were 7.49 (5.03-11.15) that of a child in DepCat 1 (most affluent). Inequalities in the prevalence of d3mft have reduced and in 2003 the odds of a child in DepCat 7 having d3mft were 4.60 (3.47-6.14) that of a child in DepCat 1. However, socio-economic inequalities in the amount of d3mft for those with d3mft have seen no reduction and have in fact increased between 1993 and 2003, with this increase approaching significance. CONCLUSION: While socio-economic inequalities in prevalence of children with d3mft have decreased in recent years, socio-economic inequalities in the amount of d3mft for those with d3mft persist. This suggests that improvements are only seen for those children with the potential for low d3mft. High d3mft persists among children from more deprived areas. The national target conceals this apparent inconsistency. PMID- 19307246 TI - A new Helsinki Declaration--but what about public health research? PMID- 19307247 TI - The relevance for Europe of the report of the Commission on the Social Determinants of Health. PMID- 19307248 TI - A vision of hope for US domestic and international health policy. PMID- 19307249 TI - Chronic diseases in Europe's migrant and ethnic minorities: challenges, solutions and a vision. AB - The pattern of chronic disease varies hugely internationally, and this is now reflected in Europe's multi-ethnic populations. This is creating challenges for epidemiology, public health and clinical care. Human rights legislation and health policies are mandating equity of service i.e. equal needs being met equally well. Indicators of race and ethnicity demonstrate important variations in health and health care, but the data are sparse, especially in Southern and Eastern Europe, and for some ethnic groups across the continent. Ethnic coding of routine health databases is required. The task will best succeed as a national one, with European level coordination and guidance on concepts. Pending this achievement, data linkage techniques can help fill the information gap. One of many ongoing debates that need resolution across Europe is on the preferred indicator of ethnicity, related terminology and mode of measurement. Original research also needs expansion, especially in relation to cohort studies and trials and boosted samples of ethnic minority groups in large scale European health surveys. Such work may require European legislation of the kind that has been effective in the United States (NIH Revitalisation Act 1993). A dialogue between policy makers, funders, researchers and practitioners is needed urgently as a precursor to engaging the public. PMID- 19307250 TI - The effect of the Irish smoke-free workplace legislation on smoking among bar workers. AB - BACKGROUND: On 29 March 2004, the Republic of Ireland (ROI) became the first EU country to introduce a nationwide ban on workplace smoking. While the focus of this measure was to protect worker health by reducing exposure to second-hand smoke, other effects such as a greater reduction in smoking prevalence and consumption were likely among bar workers. METHODS: A random sample of bar workers from Cork city were surveyed before (n = 129) and after (n = 107; 82.9% follow-up rate) implementation of the smoke-free legislation. Self report and combined self report and cotinine concentration were used to determine smoking status. For comparison a cross-sectional random telephone survey of the general population (ROI) was conducted before and 1 year after the smoke-free legislation. There were 1240 pre- and 1221 participants post-ban in the equivalent age and occupational subset of the general population. RESULTS: There was a non-significant decline in smoking prevalence among bar workers 1 year post ban (self report: -2.8% from 51.4% to 48.6%, P = 0.51; combined self report and cotinine: -4.7% from 56.1% to 51.4%, P = 0.13), but a significant decline in consumption of four cigarettes (95% CI 2.21-6.36) per day. Within the occupationally equivalent general population sub-sample there was a significant drop (3.5%, P = 0.06) in smoking prevalence but no significant change in consumption. CONCLUSIONS: Ireland's smoke-free workplace legislation was accompanied by a drop in smoking prevalence in both bar workers and the general population sub-sample. PMID- 19307251 TI - In search of the depressive self: extended medial prefrontal network during self referential processing in major depression. AB - Major depression is associated with an excessive self-focus, a tendency to engage oneself in self-referential processing. The medial frontal gyrus (MFG) is central to self-referential processing. This study aimed to explore the neural bases of this excessive self-focus and to disambiguate the role of the MFG in the pathophysiology of major depression. We presented 15 depressed patients and 15 healthy subjects with personality traits during functional magnetic resonance imaging and asked them to judge whether each trait described them ('self' condition) or a generally desirable trait ('general' condition). Both patients and healthy subjects activated the MFG in 'self' vs 'general' condition. However, the activation of the dorsal part of the MFG and of the dorsolateral prefrontal cortex (DLPFC) in 'self' vs 'general' condition was unique to patients. Additionally, patients displayed an increased functional connectivity between the MFG, the dorsal anterior cingulate cortex and the DLPFC. These results provide evidence for an extended medial prefrontal network during self-referential processing in major depression, suggesting the involvement of a greater cognitive control. PMID- 19307252 TI - Stress cardiomyopathy - a unique presentation of diabetic ketoacidosis. AB - We have documented a unique instance of diabetic ketoacidosis (DKA)-induced stress-related cardiomyopathy (CMP) in a 46-yr-old Caucasian female with type I diabetes mellitus. In times of stress, tissues with high capacity for aerobic metabolism, like myocardium, can preferentially change their metabolic substrate to ketones. The myocyte has a decreased ability to metabolize glucose and free fatty acids in stress CMP. To the best of our knowledge this is the first report of stress CMP complicating DKA. A possible mechanism for the same is hypothesized. PMID- 19307253 TI - Interpreting data in the face of competing explanations: assessing the hypothesis that observed spontaneous clearance of Helicobacter pylori was all measurement error. AB - BACKGROUND: We previously reported frequent transient positive urea breath tests for Helicobacter pylori infection in a cohort study of young children, and interpreted this as evidence of frequent spontaneous clearance of this infection. In a commentary, Perry and Parsonnet suggested that all transient positive tests we observed could be false positives and thus the appearance of transient infection could be an artifact. METHODS: We address the logic of the implicit argument that the transient infections were an artifact and we demonstrate a simple likelihood calculation to assess the plausibility of competing explanations. We calculate the likelihood of observing our data based on a range of clearance and measurement error rates and then how this updates a set of prior beliefs. RESULTS: The likelihood calculations and resulting posterior probabilities show strong support for the hypothesis of spontaneous clearance, after allowing for measurement error, even starting with a very high prior probability of no spontaneous clearance. The scenario Perry and Parsonnet present is incompatible with our data, and thus not a plausible explanation for our observations. Attributing most observed transient infections to measurement error requires assuming a high false positive rate and a very low infection rate and/or a high false negative rate, alternatives that are not supported by evidence. CONCLUSIONS: Acknowledgment of plausible levels of measurement error does not change the strong support our data provides for the hypothesis of frequent transient infection. Debate about competing explanations for observations should be accompanied by quantitative analysis that shows which is more plausible. We demonstrate one method for doing such analysis. PMID- 19307254 TI - Highly thermostable L-threonine dehydrogenase from the hyperthermophilic archaeon Thermococcus kodakaraensis. AB - l-Threonine dehydrogenase, a key enzyme in the l-threonine metabolism, catalyses the NAD(+)-dependent conversion of l-threonine to 2-amino-3-ketobutyrate, that non-enzymically decarboxylates to aminoacetone. A search of the genome sequence of hyperthermophilic archaeon, Thermococcus kodakaraensis revealed the presence of a closely related orthologue (TK0916) of archaeal and bacterial l-threonine dehydrogenase genes. Expression in Escherichia coli, purification and characterization of the TK0916 gene product revealed that this gene actually coded for a protein with high levels of l-threonine dehydrogenase activity (7.26 U mg(-1)). The enzyme exhibited highest activity at pH 12 and 90 degrees C. The K(m) values for l-threonine and NAD(+) at 50 degrees C were 1.6 mM and 0.028 mM, respectively. The enzyme activity was dependent on divalent cations. The half life of the enzyme was more than 2 h at 85 degrees C and 24 min in boiling water. This is the most thermostable threonine dehydrogenase exhibiting optimal activity at the highest pH (12) reported to date. This is the first report on the characterization of a TDH from genus Thermococcus. PMID- 19307255 TI - Cellular and mitochondrial glutathione redox imbalance in lymphoblastoid cells derived from children with autism. AB - Research into the metabolic phenotype of autism has been relatively unexplored despite the fact that metabolic abnormalities have been implicated in the pathophysiology of several other neurobehavioral disorders. Plasma biomarkers of oxidative stress have been reported in autistic children; however, intracellular redox status has not yet been evaluated. Lymphoblastoid cells (LCLs) derived from autistic children and unaffected controls were used to assess relative concentrations of reduced glutathione (GSH) and oxidized disulfide glutathione (GSSG) in cell extracts and isolated mitochondria as a measure of intracellular redox capacity. The results indicated that the GSH/GSSG redox ratio was decreased and percentage oxidized glutathione increased in both cytosol and mitochondria in the autism LCLs. Exposure to oxidative stress via the sulfhydryl reagent thimerosal resulted in a greater decrease in the GSH/GSSG ratio and increase in free radical generation in autism compared to control cells. Acute exposure to physiological levels of nitric oxide decreased mitochondrial membrane potential to a greater extent in the autism LCLs, although GSH/GSSG and ATP concentrations were similarly decreased in both cell lines. These results suggest that the autism LCLs exhibit a reduced glutathione reserve capacity in both cytosol and mitochondria that may compromise antioxidant defense and detoxification capacity under prooxidant conditions. PMID- 19307256 TI - Nailfold capillaroscopy abnormalities are associated with the presence of anti endothelial cell antibodies in Sjogren's syndrome. PMID- 19307257 TI - Perceptions of disease and health-related quality of life among patients with gout. AB - OBJECTIVE: To assess the impact of gout on health-related quality of life (HRQoL) among patients in three large US cities. METHODS: Gout patients completed the Short Form-36 (SF-36) and a series of questions regarding their gout, comorbidities and demographics. Their physicians confirmed the gout diagnosis and evaluated the severity of patient's gout. The differences in mean norm-based SF 36 scores between the US norms and gout patients and between subgroups of gout patients were calculated. The relative weight and significance of gout-related characteristics associated with patients' HRQoL were also calculated. RESULTS: The majority of the patients were males with a mean age of 62.2 years and median disease duration of 13.8 years. Most were overweight/obese with several comorbidities. Half of the patients experienced three or more gout attacks per year with a typical gout attack involving five joints and lasting for at least 4 days. The Physical Component Summary (PCS) and Mental Component Summary (MCS) was significantly lower for gout patients (P < 0.002 and P < 0.001, respectively). Among gout patients, the mean PCS and MCS were lower for those with more frequent gout attacks and greater number of affected joints (P < 0.005 and P < 0.001, respectively). After adjusting for age, gender and comorbidities, the number of joints involved during a typical and the worst gout attack had the greatest impact on patient's PCS and MCS. CONCLUSION: Gout patients had clinically significant lower HRQoL than their age-matched US norm. Comorbidities and several additional gout-related factors significantly impacted the overall HRQoL. PMID- 19307258 TI - "Public-researcher partnership". Realities in accessing records. PMID- 19307259 TI - Resting heart rate as predictor. What about absolute risks? PMID- 19307260 TI - Turning old age into a disease? On disobeying NICE guidance. PMID- 19307261 TI - Turning old age into a disease? Better evidence is the answer. PMID- 19307262 TI - Turning old age into a disease? Older people are undertreated as much as they are overtreated. PMID- 19307263 TI - Restructuring the NHS. Finsbury Health Centre's demise. PMID- 19307264 TI - Oxfam must shed its ideological bias to be taken seriously. PMID- 19307266 TI - Inquiry into food poisoning outbreak in Wales says lessons weren't learnt from past incidents. PMID- 19307268 TI - Experimental vaccine may have saved Hamburg scientist from Ebola fever. PMID- 19307269 TI - Climate change could overwhelm relief agencies, experts warn. PMID- 19307270 TI - Quality-of-life concerns differ among patients, parents, and medical providers in children and adolescents with congenital and acquired heart disease. AB - OBJECTIVES: Mortality rates in pediatric patients with heart disease have decreased dramatically in recent decades, resulting in an increasing number of survivors with morbidities that impact quality of life. The purpose of this study was to assess and compare how heart disease affects the quality of life of the pediatric cardiac patient from the perspectives of the patient, parent, and health care provider. METHOD: Individual focus groups were conducted with children (8-12 years of age) with heart disease, adolescents (13-18 years of age) with heart disease, parents of children with heart disease, parents of adolescents with heart disease, and health care providers of pediatric patients with heart disease. A structured focus group technique was used to develop a list of potential items that might affect quality of life. Participants chose the 5 most important items from the list. These items were then categorized into preidentified dimensions (physical, psychological, social, school, and other). The percentages of the total votes for all items were calculated and distributions of responses across dimensions within group and within dimension across groups were reported and compared qualitatively. RESULTS: Patient and parent groups identified similar items as important. Providers identified different items. The physical limitation item received the largest percentage of total votes in all groups (9%-20%). Analysis across dimension revealed that those items related to the physical dimension received the highest percentage of total votes among all groups (30.2%-51.2%). Analysis within dimension revealed that patients endorsed items in the physical dimension more frequently than parents or providers. Within the psychological dimension, the children selected the fewest items relative to all other groups (7.2% vs 21.3%-37.8%), whereas health care providers endorsed these items more frequently than patients or parents (child or parent of child groups 7.2%-21.3% vs health care provider group 28.8%; adolescent or parent of adolescent groups 29.6% vs health care provider group 37.8%). Differences were noted between the child and adolescent groups in the psychological (child versus adolescent: 7.2% vs 29.6%) and school (child versus adolescent: 11.2% vs 2.1%) dimensions. CONCLUSIONS: Patients and parents generally agreed on how heart disease affects the quality of life of children and adolescents, whereas health care providers had a different opinion. PMID- 19307271 TI - Repeat prenatal corticosteroid doses do not alter neonatal blood pressure or myocardial thickness: randomized, controlled trial. AB - OBJECTIVE: The goal was to determine whether repeat prenatal corticosteroid treatment alters blood pressure and myocardial wall thickness in neonates. METHODS: A randomized, double-blind, placebo-controlled trial was performed in a tertiary perinatal center. Mothers with a singleton, twin, or triplet pregnancy, at a gestational age of <32 weeks, who had received initial treatment with corticosteroid > or =7 days earlier and who were considered to be at continued risk of preterm birth were assigned randomly to receive additional weekly betamethasone or placebo treatment. One hundred forty-five infants born to 120 women were studied. Blood pressure in the first 4 weeks after birth or until hospital discharge and interventricular septal thickness and left ventricular posterior wall thickness in diastole 48 to 72 hours after birth were measured. RESULTS: There were no differences in mean, systolic, or diastolic blood pressures between infants in the placebo and repeat steroid groups. Blood pressures of infants in both groups were similar to published normal values. There were no differences between groups in interventricular septal thickness or left ventricular posterior wall thickness in diastole. In comparison with published normal ranges, however, 24% of infants had interventricular septal thickness and 32% of infants had left ventricular posterior wall thickness of >95th percentile. CONCLUSION: Exposure to repeat prenatal corticosteroid treatment did not increase neonatal blood pressure or myocardial wall thickness in infants who remained at risk of very preterm birth > or =7 days after an initial course of corticosteroid treatment. PMID- 19307272 TI - Transient ischaemic attack. PMID- 19307274 TI - HIV prevalence in US capital is at epidemic level. PMID- 19307276 TI - Israeli Bedouin villages have severe shortages of medical services. PMID- 19307277 TI - Should doctors recommend automated external defibrillators for use at home after myocardial infarction? Yes. PMID- 19307278 TI - Should doctors recommend automated external defibrillators for use at home after myocardial infarction? No. PMID- 19307280 TI - Comprehensive upstream treatment for atrial fibrillation, when and how? PMID- 19307281 TI - Reflections on reconduction after pulmonary vein isolation. PMID- 19307282 TI - Dominant frequency differences in atrial fibrillation patients with and without left ventricular systolic dysfunction. AB - AIMS: The aim of this study was to determine the mechanisms of atrial fibrillation (AF) in patients with left ventricular systolic dysfunction (LVSD). METHODS AND RESULTS: Dominant frequency (DF) spatiotemporal stability was studied in 15 patients with persistent AF (PEAF) and LVSD (Group I), 15 with PEAF without LVSD (Group II), and 10 with paroxysmal AF (PAAF) without LVSD (Group III). Dominant frequencies were analysed at 536 sites at baseline (DF1) and 26 +/- 12 min later (DF2). A DF1-DF2 difference of or= 120 ms] underwent de novo CRT (n = 336) or upgrading from RV pacing [n = 58; VVIR in 24, DDDR in 34] to CRT. The endpoint of death from any cause or major cardiovascular events, cardiovascular death or hospitalization for heart failure, and cardiovascular death or death from any cause was determined after a maximum follow-up of 7.7 years. No differences emerged between the de novo CRT and the upgrade-to-CRT groups with respect to any of the clinical endpoints. The de novo CRT and upgrade-to-CRT groups derived similar improvements in NYHA class [-1.2 vs. -1.3 (mean), both P < 0.0001), 6 min walking distance [75.9 (P < 0.0001) vs. 46.4 (P = 0.0205) m], and quality of life scores [-25.2 vs. -18.7 (both P < 0.0001)] 1 year after implantation. Response rates using a combined clinical score (>or=1 NYHA classes or >or=25% increase in 6 min walking distance plus survival with freedom from heart failure hospitalizations for 1 year) were 73.2% and 75.4%, respectively (P = NS). There were reductions in left ventricular end systolic volume [median of 20.3 mL (P = 0.0012) and 22.7 mL (P = 0.0066), respectively] and improvements in LVEF [median of 2.9% and 9.3%, respectively (both P < 0.0001)]. CONCLUSION: In patients with heart failure who are RV-paced, upgrading to CRT is associated with a similar long-term risk of mortality and morbidity to patients undergoing de novo CRT. Symptomatic improvements and degree of reverse remodelling are also comparable. PMID- 19307284 TI - Employment status and depressive symptoms in Koreans: results from a baseline survey of the Korean Longitudinal Study of Aging. AB - OBJECTIVES: We investigated the association between employment status and depressive symptoms among Korean individuals according to age and gender. METHODS: Data were obtained from a subsample of 3,907 men and 4,914 women aged 45 years and older without any severe disabilities who participated in the 2006 cross-sectional baseline survey of the Korean Longitudinal Study of Aging (KLoSA). We compared depressive symptoms as determined by the Center for Epidemiological Studies-Depression 10-item scale, adjusting for socioeconomic status, physical and cognitive function, and chronic disease. Using multiple regression analysis, we determined both age- and gender-specific differences associated with depressive symptoms and employment status. RESULTS: Employment was associated with fewer depressive symptoms among middle-aged men but not among older men. Unemployment was associated with more depressive symptoms among middle aged women but not among older women. CONCLUSION: Work role did not provide any benefit for depressive symptoms among older men and women. We discuss the increased depressive symptoms among older employed men and the differential association of employment status with age and gender in the context of Korean social structure. PMID- 19307285 TI - Why is the educational gradient of mortality steeper for men? AB - OBJECTIVES: It is often documented that the educational gradient of mortality is steeper for men than for women; yet, the explanation remains a matter of debate. We examine gender differences in the gradients within the context of marriage to determine whether overall differences reflect gender differences in health behaviors or a greater influence of men's education on spousal health. METHODS: We used data from the 1986 through 1996 National Health Interview Survey Linked Mortality Files for non-Hispanic White adults aged 55-84 years at the time of survey. We estimated Cox proportional hazards models to examine the gradients (N = 180,208). RESULTS: The educational gradient of mortality is marginally steeper for men than for women when aggregating across marital statuses; yet, this reflects a steeper gradient among unmarried men, with low-educated never married men exhibiting high levels of mortality. The gradient among unmarried men is steeper than unmarried women for causes that share smoking as a major risk factor, supporting a behavioral explanation for differences in the gradient. No gender difference in the gradient is observed for married adults. DISCUSSION: Low education and unmarried status exert a synergistic effect on men's mortality. Unmarried, low-educated men may lack social supports that encourage positive health behaviors. PMID- 19307287 TI - An introduction to artificial neural networks in bioinformatics--application to complex microarray and mass spectrometry datasets in cancer studies. AB - Applications of genomic and proteomic technologies have seen a major increase, resulting in an explosion in the amount of highly dimensional and complex data being generated. Subsequently this has increased the effort by the bioinformatics community to develop novel computational approaches that allow for meaningful information to be extracted. This information must be of biological relevance and thus correlate to disease phenotypes of interest. Artificial neural networks are a form of machine learning from the field of artificial intelligence with proven pattern recognition capabilities and have been utilized in many areas of bioinformatics. This is due to their ability to cope with highly dimensional complex datasets such as those developed by protein mass spectrometry and DNA microarray experiments. As such, neural networks have been applied to problems such as disease classification and identification of biomarkers. This review introduces and describes the concepts related to neural networks, the advantages and caveats to their use, examples of their applications in mass spectrometry and microarray research (with a particular focus on cancer studies), and illustrations from recent literature showing where neural networks have performed well in comparison to other machine learning methods. This should form the necessary background knowledge and information enabling researchers with an interest in these methodologies, but not necessarily from a machine learning background, to apply the concepts to their own datasets, thus maximizing the information gain from these complex biological systems. PMID- 19307286 TI - Socioeconomic status and age variations in health-related quality of life: results from the national health measurement study. AB - OBJECTIVES: We examine whether multiple health-related quality of life (HRQoL) measures are stratified by socioeconomic status (SES) and age in the United States. METHODS: Data are from the 2005/2006 National Health Measurement Study, a telephone survey of a nationally representative sample of U.S. adults. We plot mean HRQoL scores by SES within age groups. Regression analyses test whether education, income, and assets each have independent associations with three "preference-based" HRQoL measures and self-rated health (SRH). We test whether these associations vary by age. RESULTS: There are SES disparities in HRQoL and SRH among adults in the United States at all age groups. Income differentials in HRQoL are strong across current adult age cohorts, except the 75-89 age cohort. Education and assets have statistically significant but weaker associations with HRQoL. All three SES measures are associated with SRH (net of each other) at every age group. Those in the lowest income and education groups in the 35-44 age cohort have worse HRQoL and SRH than those in higher SES groups in the 65+ age cohort. DISCUSSION: Significant improvements in HRQoL at the population level will only be possible if we improve the HRQoL of people at the lowest end of the socioeconomic distribution. PMID- 19307288 TI - Analysis of distortion product otoacoustic emission spectra in normal-hearing adults. AB - PURPOSE: To evaluate the ability of distortion product otoacoustic emission (DPOAE) spectral characteristics to distinguish between ears with variable hearing sensitivity within the normal range. METHOD: Distortion product grams (DP grams) were acquired with primary tones decremented in 1/8-octave steps and primary-tone levels presented at 65 dB SPL (L1) and 45 dB SPL (L2) across an f2 frequency range of 842-7996 Hz from 22 normal-hearing adults (44 ears). Hearing thresholds of ears classified in Group A (n = 22) were better than ears classified in Group B (n = 22). Examined parameters of the DP-grams included spectral peak occurrence, peak height, peak width, and DPOAE levels. Analyses of variance were conducted to determine whether DP-gram parameters differed between Group A and Group B. RESULTS: For the low-resolution DP-grams examined in this study, no significant between-group differences in peak occurrence, peak height, and peak width were observed. DPOAE levels were significantly higher in ears classified in Group A compared with ears classified in Group B in individuals with symmetrical hearing. CONCLUSIONS: Although spectral peaks are evident in DP grams acquired with low resolution of the primary tones, DPOAE levels are more effective in distinguishing ears with greater hearing sensitivity from less sensitive ears. PMID- 19307289 TI - Test order effects of computerized dynamic posturography and calorics. AB - PURPOSE: To determine whether there is a test order effect of the caloric subtest of videonystagmography on the Sensory Organization Test (SOT) of computerized dynamic posturography, and to assess the learning effect of multiple posturography test sessions on the SOT score. METHOD: Scores on the 6 SOT conditions before and after caloric testing were compared for 10 participants age 18-36 years. Four SOT sessions were completed prior to caloric testing to assess the presence of a learning effect and establish precaloric baseline SOT scores. All participants had normal vestibular systems with no history of dizziness or imbalance. RESULTS: Caloric testing had a significant effect on the equilibrium score for only 1 of 6 test conditions. While meeting statistical significance, the actual change in score for this condition was very small. Equilibrium scores improved for the more difficult test conditions between the 1st and 2nd SOT sessions. CONCLUSION: Results indicate that there are minimal test order effects of calorics on the SOT. Results confirm that a significant improvement in score for more challenging test conditions occurs between SOT Sessions 1 and 2 as a result of learning. Therefore, clinicians must be cautious when using posturography to monitor patient improvement. PMID- 19307290 TI - Head shake computerized dynamic posturography in peripheral vestibular lesions. AB - PURPOSE: To determine the sensitivity of a head shake modification to the Sensory Organization Test (SOT) of dynamic posturography in identifying (a) those patients with unilateral, peripheral vestibular hypofunction as indicated by caloric irrigation findings and (b) those patients who report that head movements provoke disruption in postural control. METHOD: A prospective, single-blinded, random selection methodology was used with 91 patients stratified by the presence or absence of a significant caloric asymmetry and by the presence or absence of head movement provoked symptoms (independent variables). Postural control performance, as measured by EquiTest during the standard test and a head shake modification, served as the dependent variables. RESULTS: Receiver operating characteristic curves demonstrated only minor improvement in sensitivity (a) with the head shake modification for unilateral peripheral asymmetry and (b) for identification of those with complaints of head movement provoked imbalance. CONCLUSIONS: The head shake modification to standard SOT increased the test sensitivity to identification of patients with unilateral, peripheral vestibular hypofunction and those with head movement provoked symptoms. However, this occurred with low specificity, resulting in no significant improvement in overall performance with this head shake protocol. Suggestions for further research to improve the performance of the head shake modification of SOT for clinical application are discussed. PMID- 19307291 TI - Hearing aid outcomes for listeners with high-frequency hearing loss. AB - PURPOSE: To determine whether listeners with primarily high-frequency sensorineural hearing loss (HF SNHL) perceived benefit from amplification provided by completely-in-the-canal (CIC) hearing aids. METHOD: The Hearing Handicap for the Elderly-Screening version, the Abbreviated Profile of Hearing Aid Benefit, and the Satisfaction With Amplification in Daily Life questionnaires were mailed to 2 groups of listeners (n = 79) who matched preset criteria, including threshold at 2000 Hz and use of CIC amplification. Sixty-seven percent (n = 53) of the questionnaires were returned and were divided into 2 groups. Group I (n = 26) consisted of listeners with normal hearing through 2000 Hz, and Group II (n = 27) consisted of listeners with normal hearing only through 1000 Hz. RESULTS: Results revealed that both groups perceived significant hearing handicap, hearing aid benefit, and hearing aid satisfaction. Differences between the 2 groups on the 3 measures, however, were not statistically significant. CONCLUSIONS: Listeners with HF SNHL perceived benefit and satisfaction from amplification with CIC hearing aids. Individuals with SNHL limited to the high frequencies should be considered candidates for amplification. PMID- 19307292 TI - The activity and selectivity of fission yeast Pop2p are affected by a high affinity for Zn2+ and Mn2+ in the active site. AB - In eukaryotic organisms, initiation of mRNA turnover is controlled by progressive shortening of the poly-A tail, a process involving the mega-Dalton Ccr4-Not complex and its two associated 3'-5' exonucleases, Ccr4p and Pop2p (Caf1p). RNA degradation by the 3'-5' DEDDh exonuclease, Pop2p, is governed by the classical two metal ion mechanism traditionally assumed to be dependent on Mg(2+) ions bound in the active site. Here, we show biochemically and structurally that fission yeast (Schizosaccharomyces pombe) Pop2p prefers Mn(2+) and Zn(2+) over Mg(2+) at the concentrations of the ions found inside cells and that the identity of the ions in the active site affects the activity of the enzyme. Ion replacement experiments further suggest that mRNA deadenylation could be subtly regulated by local Zn(2+) levels in the cell. Finally, we use site-directed mutagenesis to propose a mechanistic model for the basis of the preference for poly-A sequences exhibited by the Pop2p-type deadenylases as well as their distributive enzymatic behavior. PMID- 19307293 TI - Computational and analytical framework for small RNA profiling by high-throughput sequencing. AB - The advent of high-throughput sequencing (HTS) methods has enabled direct approaches to quantitatively profile small RNA populations. However, these methods have been limited by several factors, including representational artifacts and lack of established statistical methods of analysis. Furthermore, massive HTS data sets present new problems related to data processing and mapping to a reference genome. Here, we show that cluster-based sequencing-by-synthesis technology is highly reproducible as a quantitative profiling tool for several classes of small RNA from Arabidopsis thaliana. We introduce the use of synthetic RNA oligoribonucleotide standards to facilitate objective normalization between HTS data sets, and adapt microarray-type methods for statistical analysis of multiple samples. These methods were tested successfully using mutants with small RNA biogenesis (miRNA-defective dcl1 mutant and siRNA-defective dcl2 dcl3 dcl4 triple mutant) or effector protein (ago1 mutant) deficiencies. Computational methods were also developed to rapidly and accurately parse, quantify, and map small RNA data. PMID- 19307294 TI - Tailoring RNA modular units on a common scaffold: a modular ribozyme with a catalytic unit for beta-nicotinamide mononucleotide-activated RNA ligation. AB - A novel ribozyme that accelerates the ligation of beta-nicotinamide mononucleotide (beta-NMN)-activated RNA fragments was isolated and characterized. This artificial ligase ribozyme (YFL ribozyme) was isolated by a "design and selection" strategy, in which a modular catalytic unit was generated on a rationally designed modular scaffold RNA. Biochemical analyses of the YFL ribozyme revealed that it catalyzes RNA ligation in a template-dependent manner, and its activity is highly dependent on its architecture, which consists of a modular scaffold and a catalytic unit. As the design and selection strategy was used for generation of DSL ribozyme, isolation of the YFL ribozyme indicated the versatility of this strategy for generation of functional RNAs with modular architectures. The catalytic unit of the YFL ribozyme accepts not only beta-NMN but also inorganic pyrophosphate and adenosine monophosphate as leaving groups for RNA ligation. This versatility of the YFL ribozyme provides novel insight into the possible roles of beta-NMN (or NADH) in the RNA world. PMID- 19307295 TI - Evaluation of multiple in vitro systems for assessment of CYP3A4 induction in drug discovery: human hepatocytes, pregnane X receptor reporter gene, and Fa2N-4 and HepaRG cells. AB - Prototypic CYP3A4 inducers were tested in a pregnane X receptor (PXR) reporter gene assay, Fa2N-4 cells, HepaRG cells, and primary human hepatocytes, along with negative controls, using CYP3A4 mRNA and activity endpoints, where appropriate. Over half of the compounds tested (14 of 24) were identified as time-dependent inhibitors of CYP3A4 and high mRNA/activity ratios (>10) were consistent with CYP3A4 time-dependent inhibition for compounds such as troleandomycin, ritonavir, and verapamil. Induction response was compared between two human donors; there was an excellent correlation in the EC(50) estimates (r(2) = 0.89, p < 0.001), and a weak but statistically significant correlation was noted for maximum observed induction at an optimum concentration (E(max)) (r(2) = 0.38, p = 0.001). E(max) and EC(50) estimates determined from the PXR reporter gene assay and Fa2N 4 and HepaRG cells were compared with those from hepatocytes. Overall, EC(50) values generated using hepatocytes agreed with those generated in the PXR reporter gene assay (r(2) = 0.85, p < 0.001) and Fa2N-4 (r(2) = 0.65, p < 0.001) and HepaRG (r(2) = 0.99, p < 0.001) cells. However, E(max) values generated in hepatocytes were only significantly correlated to those determined in Fa2N-4 (r(2) = 0.33, p = 0.005) and HepaRG cells (r(2) = 0.79, p < 0.001). "Gold standard" cytochrome P450 induction data can be generated using primary human hepatocytes, but a restricted, erratic supply and interdonor variability somewhat restrict routine application within a drug discovery setting. HepaRG cells are a valuable recent addition to the armory of in vitro tools for assessing CYP3A4 induction and seem to be an excellent surrogate of primary cells. PMID- 19307296 TI - Fast versus slow larval growth in an invasive marine mollusc: does paternity matter? AB - Reproductive strategies and parental effects play a major role in shaping early life-history traits. Although polyandry is a common reproductive strategy, its role is still poorly documented in relation to paternal effects. Here, we used as a case study the invasive sessile marine gastropod Crepidula fornicata, a mollusc with polyandry and extreme larval growth variation among sibling larvae. Based on paternity analyses, the relationships between paternal identity and the variations in a major early life-history trait in marine organisms, that is, larval growth, were investigated. Using microsatellite markers, paternities of 437 fast- and slow-growing larvae from 6 broods were reliably assigned to a set of 20 fathers. No particular fathers were found responsible for the specific growth performances of their offspring. However, the range of larval growth rates within a brood was significantly correlated to 1) an index of sire diversity and 2) the degree of larvae relatedness within broods. Multiple paternity could thus play an important role in determining the extent of pelagic larval duration and consequently the range of dispersal distances achieved during larval life. This study also highlighted the usefulness of using indices based on fathers' relative contribution to the progeny in paternity studies. PMID- 19307298 TI - Human ClCa1 modulates anionic conduction of calcium-dependent chloride currents. AB - Proteins of the CLCA gene family including the human ClCa1 (hClCa1) have been suggested to constitute a new family of chloride channels mediating Ca(2+) dependent Cl- currents. The present study examines the relationship between the hClCa1 protein and Ca(2+)-dependent Cl- currents using heterologous expression of hClCa1 in HEK293 and NCIH522 cell lines and whole cell recordings. By contrast to previous reports claiming the absence of Cl- currents in HEK293 cells, we find that HEK293 and NCIH522 cell lines express constitutive Ca(2+)-dependent Cl- currents and show that hClCa1 increases the amplitude of Ca(2+)-dependent Cl- currents in those cells. We further show that hClCa1 does not modify the permeability sequence but increases the Cl- conductance while decreasing the G(SCN-)/G(Cl-) conductance ratio from approximately 2-3 to approximately 1. We use an Eyring rate theory (two barriers, one site channel) model and show that the effect of hClCa1 on the anionic channel can be simulated by its action on lowering the first and the second energy barriers. We conclude that hClCa1 does not form Ca(2+)-dependent Cl- channels per se or enhance the trafficking/insertion of constitutive channels in the HEK293 and NCIH522 expression systems. Rather, hClCa1 elevates the single channel conductance of endogenous Ca(2+)-dependent Cl- channels by lowering the energy barriers for ion translocation through the pore. PMID- 19307299 TI - Neural and non-neural control of skin blood flow during isometric handgrip exercise in the heat stressed human. AB - During heat stress, isometric handgrip (IHG) exercise causes cutaneous vasoconstriction, but it remains controversial whether neural mechanisms are responsible for this observation. The objective of this study was to test the hypothesis that cutaneous vasoconstriction during IHG exercise in heat stressed individuals occurs via a neural mechanism. An axillary nerve blockade was performed to block efferent nerve traffic to the left forearm in seven healthy subjects. Two intradermal microdialysis probes were placed within forearm skin of the blocked area. Forearm skin blood flow was measured by laser-Doppler flowmetry over the microdialysis probes as well as from skin of the contralateral (unblocked) forearm. Cutaneous vascular conductance (CVC) was calculated from the ratio of skin blood flow to mean arterial pressure. Effectiveness of nerve blockade was verified by the absence of tactile sensation, as well as an absence of sweating and cutaneous vasodilatation during a whole-body heat stress. Upon this confirmation, adenosine was perfused through one of the microdialysis probes to increase skin blood flow similar to that of the unblocked site. After internal temperature increased approximately 0.7 degrees C, subjects performed 2 min of IHG exercise at 35% of maximal voluntary contraction using the non-blocked arm. IHG exercise significantly decreased CVC at the unblocked site (82.3 +/- 5.7 to 70.9 +/- 5.4%max, P = 0.005, means +/- S.E.M.) and the adenosine treated site of the blocked arm (75.2 +/- 7.2 to 68.3 +/- 6.6%max, P = 0.005), whereas CVC was unchanged at the blocked site that did not receive adenosine (15.7 +/- 2.8 to 13.7 +/- 2.0%max, P = 0.10). Importantly, the reduction in CVC was greater at the unblocked site than at the adenosine treated site (11.4 +/- 2.6 vs. 6.9 +/- 1.6%max, respectively, P = 0.01). These findings suggest that neural and non neural mechanisms contribute to the reduction in forearm CVC during IHG exercise in heat stressed humans. PMID- 19307300 TI - Endothelium-dependent vasodilatation and exercise hyperaemia in ageing humans: impact of acute ascorbic acid administration. AB - Age-related increases in oxidative stress impair endothelium-dependent vasodilatation in humans, leading to the speculation that endothelial dysfunction contributes to impaired muscle blood flow and vascular control during exercise in older adults. We directly tested this hypothesis in 14 young (22 +/- 1 years) and 14 healthy older men and women (65 +/- 2 years). We measured forearm blood flow (FBF; Doppler ultrasound) and calculated vascular conductance (FVC) responses to single muscle contractions at 10, 20 and 40% maximum voluntary contraction (MVC) before and during ascorbic acid (AA) infusion, and we also determined the effects of AA on muscle blood flow during mild (10% MVC) continuous rhythmic handgrip exercise. For single contractions, the peak rapid hyperaemic responses to all contraction intensities were impaired approximately 45% in the older adults (all P < 0.05), and AA infusion did not impact the responses in either age group. For the rhythmic exercise trial, FBF (approximately 28%) and FVC (approximately 31%) were lower (P = 0.06 and 0.05) in older versus young adults after 5 min of steady state exercise with saline. Subsequently, AA was infused via brachial artery catheter for 10 min during continued exercise. AA administration did not significantly influence FBF or FVC in young adults (1-3%; P = 0.24-0.59), whereas FBF increased 34 +/- 7% in older adults at end-exercise, and this was due to an increase in FVC (32 +/- 7%; both P < 0.05). This increase in FBF and FVC during exercise in older adults was associated with improvements in vasodilator responses to acetylcholine (ACh; endothelium dependent) but not sodium nitroprusside (SNP; endothelium independent). AA had no effect on ACh or SNP responses in the young. We conclude that acute AA administration does not impact the observed age-related impairment in the rapid hyperaemic response to brief muscle contractions in humans; however, it does significantly increase muscle blood flow during continuous dynamic exercise in older adults, and this is probably due (in part) to an improvement in endothelium-dependent vasodilatation. PMID- 19307301 TI - Progesterone signaling outside the TATA box. PMID- 19307303 TI - Integrating receptor signal inputs that influence small Rho GTPase activation dynamics at the immunological synapse. AB - The Rho GTPase Cdc42 regulates cytoskeletal changes at the immunological synapse (IS) that are critical to T-cell activation. By imaging fluorescent activity biosensors (Raichu) using fluorescence lifetime imaging microscopy, Cdc42 activation was shown to display kinetics that are conditional on the specific receptor input (through two IS-associated receptors, CD3 and beta1 integrin). CD3 triggered Cdc42 activity is dependent on the cyto-2 (NPIY) motif of the beta1 integrin cytoplasmic domain. Perturbations of the ezrin-radixin-moesin (ERM) function blocked CD3- and beta1-dependent increases in Cdc42 activity. Both IS associated receptors probably lie on a serial molecular pathway and transduce signals through the ERM-dependent machinery that is responsible for the remodeling and stabilization of the synapse. Cdc42 activity is impaired in beta1 integrin-deficient T cells that form conjugates with antigen-presenting cells but is partially restored in the context of an antigen-specific synapse. This restoration of Cdc42 activity is due, at least in part, to the recruitment and activation of beta2 integrin. PMID- 19307304 TI - G clustering is important for the initiation of transcription-induced R-loops in vitro, whereas high G density without clustering is sufficient thereafter. AB - R-loops form cotranscriptionally in vitro and in vivo at transcribed duplex DNA regions when the nascent RNA is G-rich, particularly with G clusters. This is the case for phage polymerases, as used here (T7 RNA polymerase), as well as RNA polymerases in bacteria, Saccharomyces cerevisiae, avians, mice, and humans. The nontemplate strand is left in a single-stranded configuration within the R-loop region. These structures are known to form at mammalian immunoglobulin class switch regions, thus exposing regions of single-stranded DNA for the action of AID, a single-strand-specific cytidine deaminase. R-loops form by thread-back of the RNA onto the template DNA strand, and here we report that G clusters are extremely important for the initiation phase of R-loop formation. Even very short regions with one GGGG sequence can initiate R-loops much more efficiently than random sequences. The high efficiencies observed with G clusters cannot be achieved by having a very high G density alone. Annealing of the transcript, which is otherwise disadvantaged relative to the nontemplate DNA strand because of unfavorable proximity while exiting the RNA polymerase, can offer greater stability if it occurs at the G clusters, thereby initiating an R-loop. R-loop elongation beyond the initiation zone occurs in a manner that is not as reliant on G clusters as it is on a high G density. These results lead to a model in which G clusters are important to nucleate the thread-back of RNA for R-loop initiation and, once initiated, the elongation of R-loops is primarily determined by the density of G on the nontemplate DNA strand. Without both a favorable R loop initiation zone and elongation zone, R-loop formation is inefficient. PMID- 19307305 TI - Differential cofactor requirements for histone eviction from two nucleosomes at the yeast PHO84 promoter are determined by intrinsic nucleosome stability. AB - We showed previously that the strong PHO5 promoter is less dependent on chromatin cofactors than the weaker coregulated PHO8 promoter. In this study we asked if chromatin remodeling at the even stronger PHO84 promoter was correspondingly less cofactor dependent. The repressed PHO84 promoter showed a short hypersensitive region that was flanked upstream and downstream by a positioned nucleosome and contained two transactivator Pho4 sites. Promoter induction generated an extensive hypersensitive and histone-depleted region, yielding two more Pho4 sites accessible. This remodeling was strictly Pho4 dependent, strongly dependent on the remodelers Snf2 and Ino80 and on the histone acetyltransferase Gcn5, and more weakly on the acetyltransferase Rtt109. Importantly, remodeling of each of the two positioned nucleosomes required Snf2 and Ino80 to different degrees. Only remodeling of the upstream nucleosome was strictly dependent on Snf2. Further, remodeling of the upstream nucleosome was more dependent on Ino80 than remodeling of the downstream nucleosome. Both nucleosomes differed in their intrinsic stabilities as predicted in silico and measured in vitro. The causal relationship between the different nucleosome stabilities and the different cofactor requirements was shown by introducing destabilizing mutations in vivo. Therefore, chromatin cofactor requirements were determined by intrinsic nucleosome stabilities rather than correlated to promoter strength. PMID- 19307306 TI - PLRG1 is an essential regulator of cell proliferation and apoptosis during vertebrate development and tissue homeostasis. AB - PLRG1, an evolutionarily conserved component of the spliceosome, forms a complex with Pso4/SNEV/Prp19 and the cell division and cycle 5 homolog (CDC5L) that is involved in both pre-mRNA splicing and DNA repair. Here, we show that the inactivation of PLRG1 in mice results in embryonic lethality at 1.5 days postfertilization. Studies of heart- and neuron-specific PLRG1 knockout mice further reveal an essential role of PLRG1 in adult tissue homeostasis and the suppression of apoptosis. PLRG1-deficient mouse embryonic fibroblasts (MEFs) fail to progress through S phase upon serum stimulation and exhibit increased rates of apoptosis. PLRG1 deficiency causes enhanced p53 phosphorylation and stabilization in the presence of increased gamma-H2AX immunoreactivity as an indicator of an activated DNA damage response. p53 downregulation rescues lethality in both PLRG1 deficient MEFs and zebrafish in vivo, showing that apoptosis resulting from PLRG1 deficiency is p53 dependent. Moreover, the deletion of PLRG1 results in the relocation of its interaction partner CDC5L from the nucleus to the cytoplasm without general alterations in pre-mRNA splicing. Taken together, the results of this study identify PLRG1 as a critical nuclear regulator of p53-dependent cell cycle progression and apoptosis during both embryonic development and adult tissue homeostasis. PMID- 19307307 TI - Structural and functional basis of a role for CRKL in a fibroblast growth factor 8-induced feed-forward loop. AB - The adapter protein CRKL is required for the normal development of multiple tissues that rely on fibroblast growth factor 8 (FGF8). The precise role of CRKL in receptor signaling has been unclear, however. To address this issue, we first modeled the three-dimensional structure of CRKL by molecular dynamics. By taking advantage of structural simulations, we performed in silico analysis of the interactions of the autophosphorylation sites of FGR receptor 1 (FGFR1) with the SH2 domain of CRKL or a highly related protein, CRK. As predicted by simulations, we confirm the specific physical interaction of phosphorylated Y463 (pY463) in FGFR1 with the CRKL SH2 domain at an affinity approximately 30-fold stronger than that of CRK. We also provide evidence that interactions outside of the core YXXP motif have a significant impact on physical association, which is consistent with predictions from molecular-dynamics simulations. Furthermore, we identify CRKL as an essential component of an FGF8-induced feed-forward loop permissive for efficient activation of the mitogen-activated protein kinase Erk1/2, as well as FGF8-induced anchorage-independent cell growth, using Crkl-deficient cells or a pY463 synthetic peptide. Although many cells generally require cell-matrix adhesion, our results demonstrate that CRKL permits cells to bypass the strict need for adhesion in response to FGF8 through direct interaction with receptor. PMID- 19307308 TI - Extracellular signal-regulated kinase mitogen-activated protein kinase signaling initiates a dynamic interplay between sumoylation and ubiquitination to regulate the activity of the transcriptional activator PEA3. AB - Many transcription factors are controlled through SUMO modification, and in the majority of cases this modification results in enhancements in their repressive properties. In some instances, SUMO modification and its associated repressive activities can be reversed by the action of intracellular signaling pathways, leading to enhanced transcriptional capacities of transcription factors. Here we have investigated sumoylation of the ETS domain transcription factor PEA3 and its interplay with the extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinase signaling pathway. PEA3 is modified by SUMO in vitro and in vivo on multiple sites in its N-terminal region. Activation of the ERK MAP kinase pathway promotes sumoylation of PEA3. Importantly, sumoylation of PEA3 is required for maximal activation of target gene promoters, including MMP-1 and COX 2. Molecularly, sumoylation is selectively required for synergistic activation of target gene expression with the coactivator CBP. Moreover, sumoylation of PEA3 is required for ubiquitination of PEA3 and promotes its degradation, suggesting that SUMO-mediated recycling of PEA3 plays a role in PEA3-mediated promoter activation. Thus, in contrast to the majority of other transcription factors studied, sumoylation of PEA3 plays a positive role in PEA3-mediated transcriptional activation and the ERK MAP kinase pathway cooperates with rather than antagonizes this process. PMID- 19307309 TI - Functional dynamics of Polo-like kinase 1 at the centrosome. AB - Polo-like kinase 1 (Plk1) functions as a key regulator of mitotic events by phosphorylating substrate proteins on centrosomes, kinetochores, the mitotic spindle, and the midbody. Through mechanisms that are incompletely understood, Plk1 is released from and relocalizes to different mitotic structures as cells proceed through mitosis. We used fluorescence recovery after photobleaching to examine the kinetics of this process in more detail. We observed that Plk1 displayed a range of different recovery rates that differ at each mitotic substructure and depend on both the Polo-box domain and a functional kinase domain. Upon mitotic entry, centrosomal Plk1 becomes more dynamic, a process that is directly enhanced by Plk1 kinase activity. In contrast, Plk1 displays little dynamic exchange at the midbody, a process that again is modulated by the kinase activity of Plk1. Our findings suggest that the intrinsic kinase activity of Plk1 triggers its release from early mitotic structures and its relocalization to late mitotic structures. To assess the importance of Plk1 dynamic relocalization, Plk1 was persistently tethered to the centrosome. This resulted in a G(2) delay, followed by a prominent prometaphase arrest, as a consequence of defective spindle formation and activation of the spindle checkpoint. The dynamic release of Plk1 from early mitotic structures is thus crucial for mid- to late-stage mitotic events and demonstrates the importance of a fully dynamic Plk1 at the centrosome for proper cell cycle progression. This dependence on dynamic Plk1 was further observed during the mitotic reentry of cells after a DNA damage G(2) checkpoint, as this process was significantly delayed upon centrosomal tethering of Plk1. These results indicate that mitotic progression and control of mitotic reentry after DNA damage resides, at least in part, on the dynamic behavior of Plk1. PMID- 19307310 TI - Cdk2 and Cdk4 activities are dispensable for tumorigenesis caused by the loss of p53. AB - The loss of p53 induces spontaneous tumors in mice, and p53 mutations are found in approximately 50% of human tumors. These tumors are generally caused by a number of events, including genomic instability, checkpoint defects, mitotic defects, deregulation of transcriptional targets, impaired apoptosis, and G(1) deregulation or a combination of these effects. In order to determine the role of proteins involved in G(1) control in tumorigenesis, we focused on Cdk2 and Cdk4, two cyclin-dependent kinases that in association with cyclin E and cyclin D promote the G(1)/S phase transition. We analyzed the consequence of loss of Cdk2 in p53-null animals by generating Cdk2(-/-) p53(-/-) mice. These mice are viable and developed spontaneous tumors, predominantly lymphoblastic lymphomas, similar to p53(-/-) mice. In contrast, the genotypes Cdk4(-/-) p53(-/-) were mostly lethal, with few exceptions, and Cdk2(-/-) Cdk4(-/-) p53(-/-) mice die during embryogenesis at embryonic day 13.5. To study the oncogenic potential, we generated mouse embryonic fibroblasts (MEFs) and found that p53(-/-), Cdk2(-/-) p53(-/-), Cdk4(-/-) p53(-/-), and Cdk2(-/-) Cdk4(-/-) p53(-/-) MEFs grew at similar rates without entering senescence. Ras-transformed MEFs of these genotypes were able to form colonies in vitro and induce tumors in nude mice. Our results suggest that tumorigenicity mediated by p53 loss does not require either Cdk2 or Cdk4, which necessitates considering the use of broad-spectrum cell cycle inhibitors as a means of effective anti-Cdk cancer therapy. PMID- 19307311 TI - Thoc1 deficiency compromises gene expression necessary for normal testis development in the mouse. AB - Accumulating evidence suggests that regulation of RNA processing through an RNP driven mechanism is important for coordinated gene expression. This hypothesis predicts that defects in RNP biogenesis will adversely affect the elaboration of specific gene expression programs. To explore the role of RNP biogenesis on mammalian development, we have characterized the phenotype of mice hypomorphic for Thoc1. Thoc1 encodes an essential component of the evolutionarily conserved TREX complex. TREX accompanies the elongating RNA polymerase II and facilitates RNP assembly and recruitment of RNA processing factors. Hypomorphic Thoc1 mice are viable despite significantly reduced Thoc1 expression in the tissues examined. While most tissues of Thoc1-deficient mice appear to develop and function normally, gametogenesis is severely compromised. Male infertility is associated with a loss in spermatocyte viability and abnormal endocrine signaling. We suggest that loss of spermatocyte viability is a consequence of defects in the expression of genes required for normal differentiation of cell types within the testes. A number of the genes affected appear to be direct targets for regulation by Thoc1. These findings support the notion that Thoc1 mediated RNP assembly contributes to the coordinated expression of genes necessary for normal differentiation and development in vivo. PMID- 19307312 TI - The RNA-binding protein Elavl1/HuR is essential for placental branching morphogenesis and embryonic development. AB - HuR is an RNA-binding protein implicated in a diverse array of pathophysiological processes due to its effects on the posttranscriptional regulation of AU- and U rich mRNAs. Here we reveal HuR's requirement in embryonic development through its genetic ablation. Obligatory HuR-null embryos exhibited a stage retardation phenotype and failed to survive beyond midgestation. By means of conditional transgenesis, we restricted HuR's mutation in either embryonic or endothelial compartments to demonstrate that embryonic lethality is consequent to defects in extraembryonic placenta. HuR's absence impaired the invagination of allantoic capillaries into the chorionic trophoblast layer and the differentiation of syncytiotrophoblast cells that control the morphogenesis and vascularization of the placental labyrinth and fetal support. HuR-null embryos rescued from these placental defects proceeded to subsequent developmental stages but displayed defects in skeletal ossification, fusions in limb elements, and asplenia. By coupling gene expression measurements, data meta-analysis, and HuR-RNA association assays, we identified transcription and growth factor mRNAs controlled by HuR, primarily at the posttranscriptional level, to guide morphogenesis, specification, and patterning. Collectively, our data demonstrate the dominant role of HuR in organizing gene expression programs guiding placental labyrinth morphogenesis, skeletal specification patterns, and splenic ontogeny. PMID- 19307313 TI - Molecular evolution and functional diversification of fatty acid desaturases after recurrent gene duplication in Drosophila. AB - Frequent gene duplications in the genome incessantly supply new genetic materials for functional innovation presumably driven by positive Darwinian selection. This mechanism in the desaturase gene family has been proposed to be important in triggering the pheromonal diversification in insects. With the recent completion of a dozen Drosophila genomes, a genome-wide perspective is possible. In this study, we first identified homologs of desaturase genes in 12 Drosophila species and noted that while gene duplication events are relatively frequent, gene losses are not scarce, especially in the desat1-desat2-desatF clade. By reconciling the gene tree with species phylogeny and the chromosomal synteny of the sequenced Drosophila genomes, at least one gene loss in desat2 and a minimum of six gene gains (resulting in seven desatF homologs, alpha-eta), three gene losses and one relocation in desatF were inferred. Upon branching off the ancestral desat1 lineage, both desat2 and desatF gained novel functions through accelerating protein evolution. The amino acid residues under positive selection located near the catalytic sites and the C-terminal region might be responsible for altered substrate selectivity between closely related species. The association between the expression pattern of desatF-alpha and the chemical composition of cuticular hydrocarbons implies that the ancestral function of desatF-alpha is the second desaturation at the four carbons after the first double bond in diene synthesis, and the shift from bisexual to female-specific expression in desatF-alpha occurred in the ancestral lineage of Drosophila melanogaster subgroup. A relationship between the number of expressed desatF homologs and the diene diversification has also been observed. These results suggest that the molecular diversification of fatty acid desaturases after recurrent gene duplication plays an important role in pheromonal diversity in Drosophila. PMID- 19307314 TI - Advocating for a parent with dementia in a long-term care facility: the process experienced by daughters. AB - Daughter caregivers of elders with dementia become their parents' advocates over time. This role takes on even greater importance when one or both parents are placed in a long-term care facility. This article presents the results of a qualitative study aimed at explaining how this advocacy role evolves following institutionalization. In-depth interviews were conducted with daughters (N = 14) of an institutionalized parent with dementia and selected using a theoretical sampling procedure. Data analysis using grounded theory revealed three interrelated processes that explain role transformation of the daughters: integration in the care setting, evaluation of quality of care, and development of trust. Implications for involving daughters as care partners in long-term care settings are offered. PMID- 19307315 TI - Mexican adolescents' alcohol use, family intimacy, and parent-adolescent communication. AB - Despite widespread adolescent alcohol use, research on individual and contextual factors among Mexican adolescents is limited. This study describes the relationship between adolescent risk/protective factors, parent-adolescent communication, and their effects on alcohol use of 14- to 17-year-old adolescents living in Mexico (N = 829; 458 girls, 371 boys). In this study, adolescents reported that 55% ever used alcohol, 24% used alcohol in the past 30 days, and 10% reported binge drinking. Adolescents with high family intimacy were less likely to report ever using alcohol and binge drinking. Regression analysis revealed that parent-adolescent communication mediated the effect of family intimacy on overall and binge drinking. Alcohol use prevention with Mexican adolescents should focus on family intimacy and parent-adolescent communication. PMID- 19307316 TI - Analysis of cross-sectional univariate measurements for family dyads using linear mixed modeling. AB - Outcome measurements from members of the same family are likely correlated. Such intrafamilial correlation (IFC) is an important dimension of the family as a unit but is not always accounted for in analyses of family data. This article demonstrates the use of linear mixed modeling to account for IFC in the important special case of univariate measurements for family dyads collected at a single point in time. Example analyses of data from partnered parents having a child with a chronic condition on their child's adaptation to the condition and on the family's general functioning and management of the condition are provided. Analyses of this kind are reasonably straightforward to generate with popular statistical tools. Thus, it is recommended that IFC be reported as standard practice reflecting the fact that a family dyad is more than just the aggregate of two individuals. Moreover, not accounting for IFC can affect the conclusions. PMID- 19307317 TI - Psychosocial correlates of leisure-time walking among Australian adults of lower and higher socio-economic status. AB - Adults of lower socio-economic status (SES) participate less in physical activity than those of higher SES. Understanding the correlates of physical activity participation and how these may differ between socio-economic groups can inform policies and physical activity promotion strategies. The psychosocial correlates of leisure-time walking (the most common voluntary physical activity of adults) were assessed using a survey of 2488 randomly sampled Australian adults (response rate = 74.2%). Among respondents of higher SES, there were higher levels of positive cognitions towards physical activity, and walking for leisure was more prevalent than among those of lower SES. Relationships of psychosocial attributes with leisure-time walking differed by SES. The strongest correlate of leisure time walking was perceived barriers for lower SES adults and enjoyment for those of higher SES. Social support from friends was associated with walking for both groups, while the effect of support from family was significant only for adults of lower SES. Strategies influencing leisure-time walking may have to target the specific needs of different socio-economic groups. For example, removing perceived barriers may be more appropriate to promote walking among lower SES adults. Interventions tailored for lower SES groups may help close the socio economic gap in physical activity participation. PMID- 19307318 TI - A qualitative exploration of the community partner experience in a faith-based breast cancer educational intervention. AB - Although community partner engagement is a key component in faith-based health promotion/disease prevention intervention research, the perspective of community partners on their experiences in the intervention process has been infrequently investigated. Semi-structured in-depth interviews were conducted with 12 African American community partners [i.e. four pastors and eight lay health co-ordinators (LHCs)] from eight churches in greater Baltimore, MD, USA, that engaged in a breast cancer educational intervention that followed a community-based participatory research (CBPR) approach. Audiotaped interviews were transcribed, coded and content analysis was used to identify themes across the codes. Findings show that pastors support a holistic approach to health and that LHCs act as a link between the pastors, participants and academic researchers. In addition, pastors and LHCs emphasized that the religious and/or spiritual program elements should not overpower the importance of reaching participants with critical health information regardless of their religious or spiritual beliefs. Study findings suggest that faith-based educational intervention efforts that follow a CBPR approach are important in promoting cancer awareness in the African-American community. Including community partner assessment can further elucidate critical intervention impacts and helps to address health disparities in underserved communities. PMID- 19307319 TI - African-American caregivers' breast health behavior. AB - This study utilizes a stress and coping framework which includes cognitive appraisal, personal and environmental resources, coping and stress to examine factors related to African-American caregivers' breast cancer screenings, including mammograms, clinical examinations and self-examinations. Using data from the Black Rural and Urban Caregivers Mental Health and Functioning Study, we performed separate logistic regressions for each type of breast cancer screening. Results reveal that having a regular doctor checkup (coping), care recipients having a cancer diagnosis (cognitive appraisal, and living in urban areas (environment resources) are associated with receiving a mammogram. Having greater income, having at least a high school degree (both personal resources) and having a regular doctor checkup (coping) are associated with receiving a clinical examination. Increased caregiver strain (stress), being 40 years old or older, social support (coping) and living in rural areas are associated with performing a self-examination. Targeting African-American caregivers, particularly in rural areas, for increased education on the importance of receiving breast cancer screenings is crucial to addressing health disparities. Making resources available, encouraging caregivers to get a clinical examination and a mammogram and directing public education toward caregivers are important points of intervention. PMID- 19307320 TI - Incidence of mild cognitive impairment and Alzheimer disease in Southern Brazil. AB - The objective of the study was to evaluate incident cases of Alzheimer disease (AD) and mild cognitive impairment (MCI) in an elderly community cohort in a major city of southern Brazil and to determine the variables associated with the development of cognitive dysfunction. Data were drawn from a cohort to investigate healthy aging among community elderly (N = 345) and were derived from the follow-up for a maximum of 8 years. Sociodemographic, psychiatric and medical information, the Mini-Mental State Examination (MMSE), and the Clinical Dementia Rating scale were obtained in each assessment. The Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition; DSM-IV), NINCDS-ADRDA (National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and related Disorders Association), and the Mayo Clinic criteria were applied to ascertain diagnoses of AD and MCI. The incidence rate per 1000 persons-year for MCI was 13.2 (95% confidence interval [CI] 7.79-20.91) and for AD was 14.8 (95% CI 9.04-22.94). Cognitive dysfunction was associated with education (odds ratio [OR] = 0.86; confidence limit [CL] 0.76-0.97 95%) and baseline MMSE (OR = 0.81; CL 0.70-0.94 95%). The AD incidence in this sample was higher than those reported in a previous Brazilian study. The study filled the epidemiological gap in the evaluation of MCI in Brazil. PMID- 19307321 TI - Death ideas, suicidal thoughts, and plans among nursing home residents. AB - Recent studies have shown that suicides are at least as frequent among Nursing Home (NH) residents as they are among older people in the general population. The objective of the present study was to evaluate the prevalence of death- and/or suicidal feelings, thoughts, and plans and any attempted suicides in a random, unselected sample (N=288) of individuals aged 65-years-and-over, living in NHs located in the Veneto Region (the Italian North-East). One hundred seventy-two participants were surveyed with a response rate of 59.7%. Five sample questions were asked to investigate the presence of death- and suicidal thoughts, plans and behaviors in different time periods. Among the NH residents, 30.8% admitted having had death or suicidal thoughts or plans during the month prior to the interview. The oldest-old residents (85+y) more frequently reported death-suicide ideation. This high frequency of death and/or suicidal feelings and thoughts among older NH residents should be carefully considered when planning and implementing health care programs in these facilities. PMID- 19307322 TI - Leisure activities and risk of vascular cognitive impairment in older adults. AB - "Vascular cognitive impairment'' refers to cognitive impairment caused or associated with vascular risk factors, and encompasses a cognitive spectrum ranging from mild cognitive impairment to dementia. We examined the association of leisure activity participation to risk of developing VCI in the Bronx aging study. Over 21 years, 71 of the 401 participants who were free of dementia or VCI at entry developed VCI (49 participants with VCI without dementia). We derived Cognitive and Physical Activity Scales based on frequency of leisure activity participation. A 1-point increase on the Cognitive, but not Physical Activity Scale, was associated with lower risk of VCI (hazard ratio 0.931, 95% confidence interval [CI] 0.895-0.970) in Cox analysis. Participation in cognitive but not physical leisure activities is associated with lower risk of VCI with or without dementia. Prospective studies and clinical trials are needed to define the causal role of cognitive leisure activities in influencing vascular risk for cognitive decline. PMID- 19307323 TI - Outcomes of Oregon's law mandating physician reporting of impaired drivers. AB - Oregon enacted a law in 2002 that requires some health care practitioners to report cognitively impaired drivers to the Department of Motor Vehicles. We examined reports submitted between 2003 and 2006 on 1664 potential impaired drivers. Of reported drivers, 48% were older than 80 years of age. Reports of cognitive impairment were 7 times more common than functional impairments. The most common cognitive impairments were judgment and problem solving (65%), memory (53%), and reaction time (52%). Only 10% of suspended drivers regained their driving privileges. Drivers older than 80 years of age were 6 times less likely to regain privileges compared to drivers 59 years or younger. In summary, Oregon's law resulted in loss of driving privileges in a small number of licensed drivers. Over half were aged 80 years or older, with chronic or progressive cognitive impairments. Further study is needed to determine whether this law reduces crashes and crash-related fatalities. PMID- 19307324 TI - Measuring cognition in a geriatric outpatient clinic: Rasch analysis of the Montreal Cognitive Assessment. AB - OBJECTIVE: To evaluate the psychometric properties of the Montreal Cognitive Assessment as a quantitative measure of cognitive ability. DATA ANALYZED: A total of 222 cases extracted from a clinical database (57-91 years) of patients screened for cognitive impairment in outpatient geriatric assessment clinics. DATA COLLECTED: Demographic information and individual item responses to Montreal Cognitive Assessment. RESULTS: Comparison of the data with a unidimensional Rasch model indicated that the total score obtained by summing across all items yields a reliable (0.75) quantitative estimate of global cognitive ability. All items fit the model and together spanned a range of difficulty from -3.75 to +2.88 logits. Items were assessed for differential item functioning across such patient characteristics as age, education, and language spoken. We provide a table for converting Montreal Cognitive Assessment total scores onto a linearly scaled score, with guidelines for interpreting changes in Montreal Cognitive Assessment score in terms of their statistical significance. CONCLUSIONS: The Montreal Cognitive Assessment can provide a reliable and valid quantitative estimate of cognitive ability in a geriatric cognitive disorders clinic setting. PMID- 19307325 TI - Transcriptional complexes formed by NFAT dimers regulate the induction of T cell tolerance. AB - In T cells, anergy can be induced after T cell receptor engagement in the absence of costimulation. Under these conditions, the expression of a specific set of anergy-associated genes is activated. Several lines of evidence suggest that nuclear factor of activated T cells (NFAT) proteins may regulate the expression of many of those genes; however, the nature of the complexes responsible for the induction of this new program of gene expression is unknown. Here, we show that transcriptional complexes formed by NFAT homodimers are directly responsible for the activation of at least two anergy-inducing genes, Grail and Caspase3. Our data shows that Grail expression is activated by direct binding of NFAT dimers to the Grail promoter at two different sites. Consequently, a mutant NFAT protein with impaired ability to dimerize is not able to induce an unresponsive state in T cells. Our results not only identify a new biological function for NFAT dimers but also reveal the different nature of NFAT-containing complexes that induce anergy versus those that are activated during a productive immune response. These data also establish a basis for the design of immunomodulatory strategies that specifically target each type of complex. PMID- 19307326 TI - The Wiskott-Aldrich syndrome protein is required for iNKT cell maturation and function. AB - The Wiskott-Aldrich syndrome (WAS) protein (WASp) is a regulator of actin cytoskeleton in hematopoietic cells. Mutations of the WASp gene cause WAS. Although WASp is involved in various immune cell functions, its role in invariant natural killer T (iNKT) cells has never been investigated. Defects of iNKT cells could indeed contribute to several WAS features, such as recurrent infections and high tumor incidence. We found a profound reduction of circulating iNKT cells in WAS patients, directly correlating with the severity of clinical phenotype. To better characterize iNKT cell defect in the absence of WASp, we analyzed was(-/-) mice. iNKT cell numbers were significantly reduced in the thymus and periphery of was(-/-) mice as compared with wild-type controls. Moreover analysis of was(-/-) iNKT cell maturation revealed a complete arrest at the CD44(+) NK1.1(-) intermediate stage. Notably, generation of BM chimeras demonstrated a was(-/-) iNKT cell-autonomous developmental defect. was(-/-) iNKT cells were also functionally impaired, as suggested by the reduced secretion of interleukin 4 and interferon gamma upon in vivo activation. Altogether, these results demonstrate the relevance of WASp in integrating signals critical for development and functional differentiation of iNKT cells and suggest that defects in these cells may play a role in WAS pathology. PMID- 19307327 TI - Evolution of HLA-B*5703 HIV-1 escape mutations in HLA-B*5703-positive individuals and their transmission recipients. AB - HLA-B*57 is the class I allele most consistently associated with control of human immunodeficiency virus (HIV) replication, which may be linked to the specific HIV peptides that this allele presents to cytotoxic T lymphocytes (CTLs), and the resulting efficacy of these cellular immune responses. In two HIV C clade infected populations in South Africa and Zambia, we sought to elucidate the role of HLA-B*5703 in HIV disease outcome. HLA-B*5703-restricted CTL responses select for escape mutations in three Gag p24 epitopes, in a predictable order. We show that the accumulation of these mutations sequentially reduces viral replicative capacity in vitro. Despite this, in vivo data demonstrate that there is ultimately an increase in viral load concomitant with evasion of all three HLA B*5703-restricted CTL responses. In HLA-B*5703-mismatched recipients, the previously described early benefit of transmitted HLA-B*5703-associated escape mutations is abrogated by the increase in viral load coincident with reversion. Rapid disease progression is observed in HLA-matched recipients to whom mutated virus is transmitted. These data demonstrate that, although costly escape from CTL responses can progressively attenuate the virus, high viral loads develop in the absence of adequate, continued CTL responses. These data underline the need for a CTL vaccine against multiple conserved epitopes. PMID- 19307328 TI - A role for human skin-resident T cells in wound healing. AB - Epidermal T cells have been shown to play unique roles in tissue homeostasis and repair in mice through local secretion of distinct growth factors in the skin. Human epidermis contains both alphabeta(+) and gammadelta(+) T cells whose functional capabilities are not understood. We demonstrate that human epidermal T cells are able to produce insulin-like growth factor 1 (IGF-1) upon activation and promote wound healing in a skin organ culture model. Moreover, an analysis of the functional capabilities of T cells isolated from acute versus chronic wounds revealed a striking difference. Both alphabeta(+) and Vdelta1(+) T cells isolated from acute wounds actively produced IGF-1, demonstrating that they are activated during tissue damage to participate in wound repair. In contrast, IGF-1 production could not be detected in T cells isolated from chronic wounds. In fact, skin T cells isolated from chronic wounds were refractory to further stimulation, suggesting an unresponsive state. Collectively, these results define a novel role for human epidermis-resident T cells in wound healing and provide new insight into our understanding of chronic wound persistence. PMID- 19307329 TI - Nonoperative treatment for proximal avulsion of the rectus femoris in professional American football. AB - BACKGROUND: Avulsion of the rectus femoris origin is a rare injury. The only previous report of this injury in professional American football has been limited to the kicking athlete. PURPOSE: To describe the incidence and treatment of proximal rectus femoris avulsion in the National Football League (NFL). STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The NFL Injury Surveillance System (NFLISS) was reviewed for any proximal rectus femoris avulsion injuries from 1986 to 2006, including the type and mechanism of injury, player demographics, method of treatment, and time to return to play. The NFL team physicians and trainers were surveyed as to their experience with these injuries as well. RESULTS: A total of 11 cases of proximal rectus femoris avulsion were identified starting in 1997. These injuries occurred in athletes in a variety of positions. All of these were treated nonoperatively, and the mean return to play was 69.2 days. CONCLUSION: Rectus femoris avulsions are uncommon injuries in the NFL, occurring about once a year in the entire league (once magnetic resonance imaging facilitated correct diagnosis of these injuries). Conservative treatment of these injuries usually results in return to play after 6 to 12 weeks. CLINICAL RELEVANCE: Proximal avulsions of the rectus femoris can be treated nonoperatively with a high degree of predictability for return to full, unrestricted participation in professional American football. PMID- 19307330 TI - Anterior cruciate ligament-injured subjects have smaller anterior cruciate ligaments than matched controls: a magnetic resonance imaging study. AB - BACKGROUND: Very few studies examining the predisposing anatomical factors leading to anterior cruciate ligament (ACL) injuries have examined the ACL itself, and none of these directly examined the difference in ACL properties between injured and matched control subjects. HYPOTHESIS: The ACL total volume in people who have experienced a noncontact ACL injury is smaller than that of matched controls. STUDY DESIGN: Case control study; Level of evidence, 3. METHODS: Contours of the ACL were manually identified in sagittal magnetic resonance images, and volumes were calculated for 27 contralateral, healthy knees of individuals after noncontact ACL injury and for 27 control subjects matched for gender, height, age, and weight. Validation of this method was performed on 5 porcine knees. Stepwise multiple regression was used to determine the difference in ACL volume between injured and control subjects while considering gender, height, weight, and age as potential covariates. RESULTS: Contralateral ACL volume for injured subjects was significantly smaller than for noninjured subjects (P = .0208) by 231 mm(3) after adjusting for weight, which was also a significant contributor to ACL volume (P < .0001). At the average body mass of 72.7 kg, subjects with a noncontact ACL injury had an average contralateral ACL volume of 1921 mm(3), while the corresponding control group had an average volume of 2151 mm(3). Gender, height, and age were not significant when weight was included in the regression model. CONCLUSION: This study shows that there are anthropometric differences between the knees of subjects with a noncontact ACL injury and those without an ACL injury, suggesting that ACL volume may play a direct role in noncontact ACL injury. PMID- 19307331 TI - Biomechanical evaluation of knotless anatomical double-layer double-row rotator cuff repair: a comparative ex vivo study. AB - BACKGROUND: The layered configuration of the rotator cuff tendon is not taken into account in classic rotator cuff tendon repair techniques. HYPOTHESIS: The mechanical properties of (1) the classic double-row technique, (2) a double-layer double-row (DLDR) technique in simple suture configuration, and (3) a DLDR technique in mattress suture configuration are significantly different. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty-four sheep shoulders were assigned to 3 repair groups of full-thickness infraspinatus tears: group 1, traditional double-row repair; group 2, DLDR anchor repair with simple suture configuration; and group 3, DLDR knotless repair with mattress suture configuration. After ultrasound evaluation of the repair, each specimen was cyclically loaded with 10 to 100 N for 50 cycles. Each specimen was then loaded to failure at a rate of 1 mm/s. RESULTS: There were no statistically significant differences among the 3 testing groups for the mean footprint area. The cyclic loading test revealed no significant difference among the 3 groups with regard to elongation. For the load-to-failure test, groups 2 and 3 showed no differences in ultimate tensile load when compared with group 1. However, when compared to group 2, group 3 was found to have significantly higher values regarding ultimate load, ultimate elongation, and energy absorbed. CONCLUSION: The DLDR fixation techniques may provide strength of initial repair comparable with that of commonly used double-row techniques. When compared with the knotless technique with mattress sutures, simple suture configuration of DLDR repair may be too weak. Knotless DLDR rotator cuff repair may (1) restore the footprint by the use of double-row principles and (2) enable restoration of the shape and profile. CLINICAL RELEVANCE: Double-layer double-row fixation in mattress suture configuration has initial fixation strength comparable with that of the classic double-row fixation and so may potentially improve functional results of rotator cuff repair. PMID- 19307332 TI - Lysholm score and Tegner activity level in individuals with normal knees. AB - BACKGROUND: The Lysholm score and Tegner activity scale are commonly used to document outcomes after arthroscopic knee surgery. These outcomes measurements are subjective in nature and evaluate performance and activity restrictions both before and after surgery, making them a valuable research tool when judging the effectiveness of surgical treatment. PURPOSE: To establish a normal knee data set for the Lysholm and Tegner rating systems, as well as to show how these scores are affected by age and gender. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: A subjective questionnaire that included both the Lysholm score and Tegner activity grading scale was completed by 488 subjects in the community who considered their knee function normal. Any subject reporting a history of injury or surgery was excluded from the study. The average age was 41 years (range, 18-85), with 244 men and 244 women qualifying for statistical analysis. RESULTS: The average Lysholm score was 94 (range, 43-100), and the average Tegner activity level was 5.7 (range, 1-10). The Lysholm score and age demonstrated no correlation. The Tegner activity level was inversely correlated with age. The average Tegner activity level for men was 6.0, and the average activity scale for women was 5.4. There was no significant difference in the Lysholm score between men and women. CONCLUSION: These data acquired from a normal, healthy population provide a standard point of reference for the injured or postsurgical knee. These data also serve as ideal tools when counseling patients about realistic expectations after surgery, based on age and gender. PMID- 19307333 TI - How long should acute anterior dislocations of the shoulder be immobilized in external rotation? AB - BACKGROUND: Immobilization of the shoulder in external rotation has been shown to reduce the risk of recurrence after traumatic anteroinferior shoulder dislocation. It remains unclear how duration of immobilization affects labral coaptation. HYPOTHESIS: Immobilization of the shoulder in 30 degrees of external rotation for 5 weeks allows better coaptation of the anteroinferior labrum than does an immobilization period of 3 weeks. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Twenty-two patients with traumatic anteroinferior dislocation of the glenohumeral joint were included in this study. Patients were divided into 2 groups. Group 1 consisted of the initial 11 patients (mean age, 37.4 years) immobilized for 3 weeks; group 2 consisted of the subsequent 11 patients (mean age, 29.7 years) immobilized for 5 weeks in 30 degrees of external rotation. With use of magnetic resonance imaging, displacement and separation of the glenoid labrum and anterior joint effusion were assessed in different arm positions (internal rotation, neutral rotation, 30 degrees of external rotation, maximum external rotation) within 3 days, 3 weeks, and 5 weeks after reduction. RESULTS: Displacement and separation of the labrum and anterior joint effusion were significantly less, particularly with maximum external rotation compared with neutral and internal rotation, during the acute magnetic resonance imaging evaluation in both groups (P < .05). No statistically significant differences were found in all parameters comparing internal rotation with neutral rotation, 30 degrees of external rotation, and maximum external rotation in both groups after 5 weeks (P > .05). No statistically significant differences were found between both groups comparing the results of the measured variables during the acute, 3-week, and 5-week magnetic resonance imaging examinations (P > .05). CONCLUSION: Immobilization of the shoulder in 30 degrees of external rotation seems to allow a similar coaptation of the glenoid labrum, regardless of duration of immobilization (3 vs 5 weeks). Clinical trials are needed to evaluate the effect of these results on recurrence rates. The optimum position of immobilization in external rotation has yet to be determined. PMID- 19307334 TI - Sexual behaviour change in countries with generalised HIV epidemics? Evidence from population-based cohort studies in sub-Saharan Africa. PMID- 19307335 TI - Trends in age at first sex in Uganda: evidence from Demographic and Health Survey data and longitudinal cohorts in Masaka and Rakai. AB - OBJECTIVES: To derive the best possible estimates of trends in age at first sex (AFS) among successive cohorts of Ugandan men and women based on all the data available from the Demographic and Health Surveys (DHS) and cohort studies in Masaka and Rakai districts. METHODS: The datasets from the DHS, Masaka cohort and Rakai cohort were analysed separately. Survival analysis methods were used to estimate median AFS for men and women born in the 1950s-1980s and to compute hazard ratios for first sex, comparing later cohorts with earlier cohorts. RESULTS: The DHS and Masaka data showed an increase in AFS in women in the more recent birth cohorts compared with those born before 1970, but this was less apparent in the Rakai data. Successive male cohorts in Masaka appeared first to have an increased AFS which subsequently decreased, a trend that was also apparent (but not significant) in the DHS data. Younger men in Rakai had an earlier AFS than those born before 1980. CONCLUSIONS: Women in Uganda who were born after 1970 have, on average, had sex at a later age than those born earlier. For men, AFS has not changed consistently over the period in question. Differences between Masaka and Rakai may reflect socioeconomic differences. Most of the change in AFS occurred too late to have contributed to the initial decline in the incidence of HIV. PMID- 19307336 TI - Influence of timing of sexual debut and first marriage on sexual behaviour in later life: findings from four survey rounds in the Kisesa cohort in northern Tanzania. AB - OBJECTIVES: To evaluate quality of sexual debut and first marriage data, measure trends and study the association of risky sexual behaviour in youth with adult risk behaviour. METHODS: Reports on age at first sex (AFS) and age at first marriage (AFM) from the Kisesa cohort study, 1994-2004, were evaluated for consistency and used to describe trends in median age-at-event and time spent single but sexually active in different birth cohorts. The association of these variables with marital stability and numbers of partners at later ages was explored using statistical regression techniques. RESULTS: AFS and AFM were inconsistently reported by 32% and 33% of respondents, respectively, but there was no general tendency to report lower or higher ages at a later report date. In 10-year birth cohorts born between 1950-9 and 1980-9, male median AFS declined from 18.1 to 17.0 years and female median AFM rose from 16.2 to 16.6 years. Young people of both sexes currently spend longer sexually active but unmarried than previously. Early marriage is statistically associated with remarriage and polygamy; longer time between sexual debut and marriage is associated with higher numbers of partners at later stages of life. CONCLUSION: Inconsistent reporting of age-at-event introduces noise but does not bias estimates of population level indicators. Lengthening time spent single and sexually active suggests that men and women entering first marriage will have been exposed to increased numbers of non-marital partners. Successful youth interventions may also influence adult behaviour. PMID- 19307337 TI - Sexual and marital trajectories and HIV infection among ever-married women in rural Malawi. AB - OBJECTIVE: To explore how sexual and marital trajectories are associated with HIV infection among ever-married women in rural Malawi. METHODS: Retrospective survey data and HIV biomarker data for 926 ever-married women interviewed in the Malawi Diffusion and Ideational Change Project were used. The associations between HIV infection and four key life course transitions considered individually (age at sexual debut, premarital sexual activity, entry into marriage and marital disruption by divorce or death) were examined. These transitions were then sequenced to construct trajectories that represent the variety of patterns in the data. The association between different trajectories and HIV prevalence was examined, controlling for potentially confounding factors such as age and region. RESULTS: Although each life course transition taken in isolation may be associated with HIV infection, their combined effect appeared to be conditional on the sequence in which they occurred. Although early sexual debut, not marrying one's first sexual partner and having a disrupted marriage each increased the likelihood of HIV infection, their risk was not additive. Women who both delayed sexual debut and did not marry their first partner are, once married, more likely to experience marital disruption and to be HIV-positive. Women who marry their first partner but who have sex at a young age, however, are also at considerable risk. CONCLUSIONS: These findings identify the potential of a life course perspective for understanding why some women become infected with HIV and others do not, as well as the differentials in HIV prevalence that originate from the sequence of sexual and marital transitions in one's life. The analysis suggests, however, the need for further data collection to permit a better examination of the mechanisms that account for variations in life course trajectories and thus in lifetime probabilities of HIV infection. PMID- 19307338 TI - Is sexual risk taking behaviour changing in rural south-west Uganda? Behaviour trends in a rural population cohort 1993-2006. AB - OBJECTIVE: To describe sexual behaviour trends in a rural Ugandan cohort in the context of an evolving HIV epidemic, 1993-2006. METHODS: Sexual behaviour data were collected annually from a population cohort in which HIV serological surveys were also conducted. Behaviour trends were determined using survival analysis and logistic regression. Trends are reported based on the years in which the respective indicators were collected. RESULTS: Between 1993 and 2006, median age at first sex increased from 16.7 years to 18.2 years among 17-20-year-old girls and from 18.5 years to 19.9 years among boys. Both sexes reported a dip in age at sexual debut between 1998 and 2001. One or more casual partners in the past 12 months among men rose from 11.6% in 1997 to 12.7% in 2004 and then declined to 10.2% in 2006. Among women it increased from 1.4% in 1997 to 3.7% in 2004 and then reduced to 1.4% in 2006. The rise in casual partners between 1997 and 2004 was driven mainly by older age groups. Trends in condom use with casual partners varied by age, increasing among those aged 35+ years, declining in the middle age groups and presenting a dip and then a rise in the youngest aged group (13-19 years). CONCLUSION: Among youth, risky behaviour declined but increased in the late 1990s/early 2000s. Among those aged 35+ years, condom use rose but casual partners also rose. Several indicators portrayed a temporary increase in risk taking behaviour from 1998 to 2002. PMID- 19307339 TI - Measuring trends in age at first sex and age at marriage in Manicaland, Zimbabwe. AB - OBJECTIVE: To identify reporting biases and to determine the influence of inconsistent reporting on observed trends in the timing of age at first sex and age at marriage. METHODS: Longitudinal data from three rounds of a population based cohort in eastern Zimbabwe were analysed. Reports of age at first sex and age at marriage from 6837 individuals attending multiple rounds were classified according to consistency. Survival analysis was used to identify trends in the timing of first sex and marriage. RESULTS: In this population, women initiate sex and enter marriage at younger ages than men but spend much less time between first sex and marriage. Among those surveyed between 1998 and 2005, median ages at first sex and first marriage were 18.5 years and 21.4 years for men and 18.2 years and 18.5 years, respectively, for women aged 15-54 years. High levels of reports of both age at first sex and age at marriage among those attending multiple surveys were found to be unreliable. Excluding reports identified as unreliable from these analyses did not alter the observed trends in either age at first sex or age at marriage. Tracing birth cohorts as they aged revealed reporting biases, particularly among the youngest cohorts. Comparisons by birth cohorts, which span a period of >40 years, indicate that median age at first sex has remained constant over time for women but has declined gradually for men. CONCLUSIONS: Although many reports of age at first sex and age at marriage were found to be unreliable, inclusion of such reports did not result in artificial generation or suppression of trends. PMID- 19307340 TI - Role of widows in the heterosexual transmission of HIV in Manicaland, Zimbabwe, 1998-2003. AB - BACKGROUND: AIDS is the main driver of young widowhood in southern Africa. METHODS: The demographic characteristics of widows, their reported risk behaviours and the prevalence of HIV were examined by analysing a longitudinal population-based cohort of men and women aged 15-54 years in Manicaland, eastern Zimbabwe. The results from statistical analyses were used to construct a mathematical simulation model with the aim of estimating the contribution of widow behaviour to heterosexual HIV transmission. RESULTS: 413 (11.4%) sexually experienced women and 31 (1.2%) sexually experienced men were reported to be widowed at the time of follow-up. The prevalence of HIV was exceptionally high among both widows (61%) and widowers (male widows) (54%). Widows were more likely to have high rates of partner change and engage in a pattern of transactional sex than married women. Widowers took partners who were a median of 10 years younger than themselves. Mathematical model simulations of different scenarios of sexual behaviour of widows suggested that the sexual activity of widow(er)s may underlie 8-17% of new HIV infections over a 20-year period. CONCLUSIONS: This combined statistical analysis and model simulation suggest that widowhood plays an important role in the transmission of HIV in this rural Zimbabwean population. High-risk partnerships may be formed when widowed men and women reconnect to the sexual network. PMID- 19307341 TI - Age at first sex in rural South Africa. AB - OBJECTIVES: To identify factors associated with sexual debut and early age at first sex (AFS) among young men and women (12-25 years) in a population with a high prevalence and incidence of HIV in rural South Africa. METHODS: Longitudinal data from four rounds (2003-7) of a prospective population-based HIV and sexual behaviour survey in rural KwaZulu-Natal were used to investigate the distribution and predictors of earlier first sex. Survival analyses were used, and each analysis considered men and women separately. RESULTS: Among the 4724 women and 4029 men who were virgins at the beginning of the period, the median AFS was 18.5 and 19.2 years, respectively. In multivariable models, factors associated with earlier AFS across gender were periurban residence (vs rural), ever use of alcohol and knowing at least one person who had HIV, while school attendance had a significant protective effect. Other factors were important for one gender only. Maternal death was significantly associated with earlier AFS for women, in the same way that paternal death was for young men, while mother's membership of the same household significantly delayed AFS of young men. The analysis of early first sex confirmed the same factors to be important as in the overall analyses for men and women. CONCLUSION: Given the association of individual, household and community level factors with sexual debut, a multisectorial approach to prevention and targeting in youth programmes is recommended. PMID- 19307342 TI - Comparative assessment of the quality of age-at-event reporting in three HIV cohort studies in sub-Saharan Africa. AB - OBJECTIVES: To assess inconsistencies in reported age at first sex (AFS) and age at first marriage (AFM) in three African cohorts, and consider their implications for interpreting trends in sexual and marital debut. METHODS: Data were analysed from population-based cohort studies in Zimbabwe, Uganda and South Africa with 3, 10 and 4 behavioural survey rounds, respectively. Three rounds over a similar time frame were selected from each site for comparative purposes. The consistency of AFS and AFM reports was assessed for each site by comparing responses made by participants in multiple surveys. Respondents were defined as unreliable if less than half of all their age-at-event reports were the same. Kaplan-Meier functions were used to describe the cumulative proportion (1) having had sex and (2) married by age, stratified by sex, birth cohort and site, to compare the influence of reporting inconsistencies on these estimates. RESULTS: Among participants attending all three comparable rounds, the percentage with unreliable AFS reports ranged from 30% among South African women to 56% among Zimbabwean men, with similar patterns observed for AFM. Inclusion of unreliable reports had little effect on estimates of median age-at-event in all sites. There was some evidence from the 1960-9 birth cohort that women in Uganda and both sexes in South Africa reported later AFS as they aged. CONCLUSION: Although reporting quality is unlikely to affect comparisons of AFS and AFM between settings, care should be taken not to overinterpret small changes in reported age at-event over time within each site. PMID- 19307343 TI - Trends in marriage and time spent single in sub-Saharan Africa: a comparative analysis of six population-based cohort studies and nine Demographic and Health Surveys. AB - OBJECTIVES: To describe trends in age at first sex (AFS), age at first marriage (AFM) and time spent single between events and to compare age-specific trends in marital status in six cohort studies. METHODS: Cohort data from Uganda, Tanzania, South Africa, Zimbabwe and Malawi and Demographic and Health Survey (DHS) data from Uganda, Tanzania and Zimbabwe were analysed. Life table methods were used to calculate median AFS, AFM and time spent single. In each study, two surveys were chosen to compare marital status by age and identify changes over time. RESULTS: Median AFM was much higher in South Africa than in the other sites. Between the other populations there were considerable differences in median AFS and AFM (AFS 17-19 years for men and 16-19 years for women, AFM 21-24 years and 18-19 years, respectively, for the 1970-9 birth cohort). In all surveys, men reported a longer time spent single than women (median 4-7 years for men and 0-2 years for women). Median years spent single for women has increased, apart from in Manicaland. For men in Rakai it has decreased slightly over time but increased in Kisesa and Masaka. The DHS data showed similar trends to those in the cohort data. The age specific proportion of married individuals has changed little over time. CONCLUSIONS: Median AFS, AFM and time spent single vary considerably among these populations. These three measures are underlying determinants of sexual risk and HIV infection, and they may partially explain the variation in HIV prevalence levels between these populations. PMID- 19307344 TI - Reported number of sexual partners: comparison of data from four African longitudinal studies. AB - OBJECTIVE: To compare reported numbers of sexual partners in Eastern and Southern Africa. METHODS: Sexual partnership data from four longitudinal population-based surveys (1998-2007) in Zimbabwe, Uganda and South Africa were aggregated and overall proportions reporting more than one lifetime sexual partner calculated. A lexis-style table was used to illustrate the average lifetime sexual partners by site, sex, age group and birth cohort. The male-to-female ratio of mean number of partnerships in the last 12 months was calculated by site and survey. For each single year of age, the proportion sexually active in the past year, the mean number of partners in the past year and the proportion with more than one partner in the past year were calculated. RESULTS: Over 90% of men and women between 25 and 45 years of age reported being sexually active during the past 12 months, with most reporting at least one sexual partner. Overall, men reported higher numbers of lifetime sexual partners and partners in the last year than women. The male-to-female ratio of mean partnerships in the last year ranged from 1.41 to 1.86. In southern African cohorts, individuals in later birth cohorts reported fewer sexual partners and a lower proportion reported multiple partnerships compared with earlier birth cohorts, whereas these behavioural changes were not observed in the Ugandan cohorts. Across the four sites, reports of sexual partnerships followed a similar pattern for each sex. CONCLUSIONS: The longitudinal results show that reductions in the number of partnerships were more evident in southern Africa than in Uganda. PMID- 19307345 TI - STIs and HIV in Pakistan: from analysis to action. PMID- 19307346 TI - Prevalence of HIV, HCV and sexually transmitted infections among injecting drug users in Rawalpindi and Abbottabad, Pakistan: evidence for an emerging injection related HIV epidemic. AB - OBJECTIVES: To measure the prevalence of hepatitis C virus (HCV), HIV and sexually transmitted infections (STI) among injecting drug users (IDUs) in Rawalpindi and Abbottabad and to examine risk factors associated with HIV and HCV. METHODS: Two cross-sectional surveys were performed of community-recruited IDUs with collection of clinical specimens for testing of HCV, HIV and other STIs. Behavioural data were collected through interviewer-administered questionnaires. Characteristics and risk behaviours were compared across cities. Univariate and multivariate analyses explored risk factors associated with HIV and HCV. RESULTS: The prevalence of HIV was 2.6% (95% CI 0.83% to 4.5%) in Rawalpindi (n = 302) and zero in Abbottabad (n = 102). The prevalence of HCV was significantly higher in Rawalpindi at 17.3% (95% CI 13.0% to 21.6%) than in Abbottabad at 8% (95% CI 2.6% to 13.4%). The prevalence of other STIs was low in both cities, with <2% of participants having current gonorrhoea or Chlamydia and <3% with active syphilis. Injecting risk behaviours were greater in Rawalpindi. An increased risk of HCV was associated with using informal sources as a main source of new needles/syringes (OR 2.8, 95% CI 1.3 to 6.0) compared with pharmacies and a history of drug treatment (OR 3.7, 95% CI 0.9 to 11.6). Reporting symptoms of an STI was associated with decreased odds of HIV in Rawalpindi (OR 0.02, 95% CI 0.03 to 0.9). CONCLUSIONS: The findings suggest recent transmission of HIV and HCV and point to the urgent need for the provision of clean needles/syringes to IDUs and a review of how needles/syringes are currently provided via healthcare establishments. PMID- 19307347 TI - Modelling the transmission of HIV and HCV among injecting drug users in Rawalpindi, a low HCV prevalence setting in Pakistan. AB - BACKGROUND: In 2007, a survey in Rawalpindi found the prevalence of hepatitis C virus (HCV) in injecting drug users (IDUs) to be low (17%), despite widespread needle/syringe sharing. This analysis uses modelling to explore hypotheses for the low prevalence of HCV to project the future HIV/HCV epidemic and to estimate the impact of a generic intervention. METHODS: An HIV/HCV transmission model was developed and parameterized using data from Rawalpindi. By incorporating different assumptions about the relative frequency/importance of needle/syringe sharing events among "strangers" and people they have shared with before, and undertaking extensive uncertainty analyses to fit the model for each scenario, the validity of different hypotheses for the low HCV prevalence was determined. Model fits were used to project the future HIV/HCV epidemic and the impact of reducing needle/syringe sharing among different IDU subgroups. RESULTS: The model projections suggest that the low HCV prevalence in Rawalpindi is probably due to most HIV/HCV transmissions occurring in a small IDU subgroup that shares needles/syringes frequently with strangers, with most needle/syringe sharing incidents being low risk. Projections suggest that the prevalence of HIV in IDUs will increase to 5-12% by 2015, and the prevalence of HCV will increase if HIV increases HCV transmission. Moderate reductions in needle/syringe sharing (>40%) could reduce the number of HCV/HIV infections (approximately 45%) if all IDUs are reached, although less impact is achieved if high-risk IDUs are not reached. CONCLUSIONS: Despite many needle/syringe sharing events possibly being low risk in Rawalpindi, the model projects that the prevalence of HIV/HCV in IDUs is likely to increase. This highlights the importance of intervening in this low prevalence setting. PMID- 19307348 TI - Understanding the context of male and transgender sex work using peer ethnography. AB - OBJECTIVES: To distinguish between three distinct groups of male and transgender sex workers in Pakistan and to demonstrate how members of these stigmatized groups need to be engaged in the research process to go beyond stated norms of behaviour. METHODS: A peer ethnography study was undertaken in a major city in Pakistan. 15 male and 15 transgender sex workers were trained as peer researchers to each interview three peers in their network. Analysis was based on interviews with peer researchers as well as observation of dynamics during training and analysis workshops. RESULTS: The research process revealed that, within the epidemiological category of biological males who sell sex, there are three sociologically different sexual identities: khusras (transgender), khotkis (feminized males) and banthas (mainstream male identity). Both khusras and khotkis are organised in strong social structures based on a shared identity. While these networks provide emotional and material support, they also come with rigid group norms based on expected "feminine" behaviours. In everyday reality, sex workers showed fluidity in both behaviour and identity according to the situational context, transgressing both wider societal and group norms. The informal observational component in peer ethnography was crucial for the accurate interpretation of interview data. Participant accounts of behaviour and relationships are shaped by the research contexts including who interviews them, at what stage of familiarity and who may overhear the conversation. CONCLUSIONS: To avoid imposing a "false clarity" on categorisation of identity and assumed behaviour, it is necessary to go beyond verbal accounts to document the fluidity of everyday reality. PMID- 19307349 TI - Protecting the unprotected: mixed-method research on drug use, sex work and rights in Pakistan's fight against HIV/AIDS. AB - OBJECTIVES: To investigate the nature and extent of human rights abuses against three vulnerable groups (injecting drug users (IDUs) and male and female sex workers), to understand the social and sexual linkages between them and to examine how protecting their rights could enhance the impact of HIV prevention policies. METHODS: In-depth interviews were carried out with 38 high-risk respondents (IDUs and female, male and transgender sex workers) and a bio behavioural survey was performed of 813 IDU/sex worker respondents in Rawalpindi. RESULTS: People in all vulnerable groups interacted both sexually and socially. All groups experienced human rights abuses by state and non-state actors which increased their HIV risk. Non-state actors, including relations and sex worker clients, are responsible for verbal, physical and sexual violence. State actors (particularly police) perpetrate harassment, exploitation and abuse of all vulnerable groups with impunity. Health service providers fail to provide adequate services for vulnerable groups. CONCLUSIONS: High levels of discrimination and abuse of human dignity of all groups studied were revealed. This violates their physical and mental integrity and also leads to an increased risk of HIV. The sexual and social interactions between groups mean that human rights abuses experienced by one high-risk group can increase the risk of HIV both for them and other groups. The protection of human rights needs to become an integral part of a multisector response to the risk of HIV/AIDS by state and non state agencies. The Government of Pakistan should work at both legal and programme levels to protect the rights of, and minimise discrimination against, groups vulnerable to HIV in order to reduce the potential for the spread of HIV before the epidemic takes hold. PMID- 19307350 TI - Political feasibility of scaling-up five evidence-informed HIV interventions in Pakistan: a policy analysis. AB - BACKGROUND: Drawing on policy theories, an assessment was made of the perceived political feasibility of scaling-up five evidence-based interventions to curb Pakistan's HIV epidemic: needle and syringe exchange programmes; targeted behaviour change communication; sexual health care for male and transgender sex workers; sexual and reproductive health care for female sex workers; and promoting and protecting the rights of those at greatest risk. METHOD: A questionnaire was emailed to 40 stakeholders and completed by 22. They expressed their level of agreement with 15 statements for each intervention (related to variables associated with policy success). Semi-structured interviews were conducted with 12 respondents. RESULTS: The interventions represent considerable change from the status quo, but are perceived to respond to widely acknowledged problems. These perceptions, held by the HIV policy elite, need to be set in the context of the prevailing view that the AIDS response is not warranted given the small and concentrated nature of the epidemic and that the interventions do not resonate closely with values held by society. The interventions were perceived to be evidence-based, supported by at least one donor and subject to little resistance from frontline staff as they will be implemented by contracted non government organisations. The results were mixed in terms of other factors determining political feasibility, including the extent to which interventions are easy to explain, exhibit simple technical features, require few additional funds, are supported and not opposed by powerful stakeholders. CONCLUSION: The interventions stand a good chance of being implemented although they depend on donor support. The prospects for scaling them would be improved by ongoing policy analysis and strengthening of domestic constituencies among the target groups. PMID- 19307351 TI - HIV and other sexually transmitted infections among men, transgenders and women selling sex in two cities in Pakistan: a cross-sectional prevalence survey. AB - OBJECTIVES: The extent and possibilities of spread of the HIV epidemic are not fully understood in Pakistan. A survey was conducted among men, women and transgender populations selling sex in Rawalpindi (Punjab) and Abbottabad (North West Frontier Province) in order to inform evidence-based programme planning. METHODS: A cross-sectional survey was performed with participants recruited through respondent-driven sampling. Male and transgender sex workers were analysed in three gender groups; women were analysed as one group. Behavioural surveys were conducted and clinical specimens collected. Laboratory tests looked for evidence of acute infection (gonorrhoea, Chlamydia, syphilis, Trichomonas) and infection over the lifetime (HIV, herpes simplex virus-2, syphilis). Predictors of infection were explored using univariable and multivariable logistic regression. RESULTS: The prevalence of HIV was low in 917 male and transgender sex workers and absent in 533 female sex workers in the study. High levels of current sexually transmitted infections were found, predominantly among transgender sex workers. Risk behaviours were common and knowledge of HIV was extremely low. Multivariable analysis found a large number of factors associated with higher levels of infection, including experience of forced first sex. Protection against risk was low, but those sex workers who reported using condoms at last sex had lower rates of infection. CONCLUSIONS: The HIV epidemic is currently in its early stages among people who sell sex, but there may be potential for a much greater spread given the levels of other sexually transmitted infections found and the concomitant low levels of both protective knowledge and risk-reducing behaviours. Action is needed now to avert an epidemic. Framing interventions by upholding the recognition and protection of human rights is vital. PMID- 19307352 TI - Considering the patient in patient-delivered partner therapy. PMID- 19307354 TI - Long-term follow-up trial of oral rifampin-cotrimoxazole combination versus intravenous cloxacillin in treatment of chronic staphylococcal osteomyelitis. AB - Oral therapies alternative to fluoroquinolones against staphylococcal chronic osteomyelitis have not been evaluated in comparative studies. Consecutive nonaxial Staphylococcus aureus chronic osteomyelitis cases were included in a comparative trial after debridement. Fifty patients were randomized: group A (n = 22) was treated with cloxacillin for 6 weeks intravenously plus 2 weeks orally (p.o.), and group B (n = 28) was treated with rifampin-cotrimoxazole for 8 weeks p.o. During follow-up (10 years), five relapses occurred: two (10%) in group A and three (11%) in group B. Foreign-body maintenance was associated with relapse (P = 0.016). Oral rifampin-cotrimoxazole treatment showed outcomes comparable to those for intravenous cloxacillin treatment. PMID- 19307355 TI - Population pharmacokinetics of ganciclovir in solid-organ transplant recipients receiving oral valganciclovir. AB - Valganciclovir (VGC) is an oral prodrug of ganciclovir (GCV) recently introduced for prophylaxis and treatment of cytomegalovirus infection. Optimal concentration exposure for effective and safe VGC therapy would require either reproducible VGC absorption and GCV disposition or dosage adjustment based on therapeutic drug monitoring (TDM). We examined GCV population pharmacokinetics in solid organ transplant recipients receiving oral VGC, including the influence of clinical factors, the magnitude of variability, and its impact on efficacy and tolerability. Nonlinear mixed effect model (NONMEM) analysis was performed on plasma samples from 65 transplant recipients under VGC prophylaxis or treatment. A two-compartment model with first-order absorption appropriately described the data. Systemic clearance was markedly influenced by the glomerular filtration rate (GFR), patient gender, and graft type (clearance/GFR = 1.7 in kidney, 0.9 in heart, and 1.2 in lung and liver recipients) with interpatient and interoccasion variabilities of 26 and 12%, respectively. Body weight and sex influenced central volume of distribution (V(1) = 0.34 liter/kg in males and 0.27 liter/kg in females [20% interpatient variability]). No significant drug interaction was detected. The good prophylactic efficacy and tolerability of VGC precluded the demonstration of any relationship with GCV concentrations. In conclusion, this analysis highlights the importance of thorough adjustment of VGC dosage to renal function and body weight. Considering the good predictability and reproducibility of the GCV profile after treatment with oral VGC, routine TDM does not appear to be clinically indicated in solid-organ transplant recipients. However, GCV plasma measurement may still be helpful in specific clinical situations. PMID- 19307356 TI - Bone penetration of amoxicillin and clavulanic acid evaluated by population pharmacokinetics and Monte Carlo simulation. AB - Amoxicillin (amoxicilline)-clavulanic acid has promising activity against pathogens that cause bone infections. We present the first evaluation of the bone penetration of a beta-lactam by population pharmacokinetics and pharmacodynamic profiling via Monte Carlo simulations. Twenty uninfected patients undergoing total hip replacement received a single intravenous infusion of 2,000 mg/200 mg amoxicillin-clavulanic acid before surgery. Blood and bone specimens were collected. Bone samples were pulverized under liquid nitrogen with a cryogenic mill, including an internal standard. The drug concentrations in serum and total bone were analyzed by liquid chromatography-tandem mass spectrometry. We used NONMEM and S-ADAPT for population pharmacokinetic analysis and a target time of the non-protein-bound drug concentration above the MIC for > or = 50% of the dosing interval for near-maximal bactericidal activity in serum. The median of the ratio of the area under the curve (AUC) for bone/AUC for serum was 20% (10th to 90th percentile for between-subject variability [variability], 16 to 25%) in cortical bone and 18% (variability, 11 to 29%) in cancellous bone for amoxicillin and 15% (variability, 11 to 21%) in cortical bone and 10% (variability, 5.1 to 21%) in cancellous bone for clavulanic acid. Analysis in S-ADAPT yielded similar results. The equilibration half-lives between serum and bone were 12 min for amoxicillin and 14 min for clavulanic acid. For a 30-min infusion of 2,000 mg/200 mg amoxicillin-clavulanic acid every 4 h, amoxicillin achieved robust (> or = 90%) probabilities of target attainment (PTAs) for MICs of < or = 12 mg/liter in serum and 2 to 3 mg/liter in bone and population PTAs above 95% against methicillin-susceptible Staphylococcus aureus in bone and serum. The AUC of amoxicillin-clavulanic acid was 5 to 10 times lower in bone than in serum, and amoxicillin-clavulanic acid achieved a rapid equilibrium and favorable population PTAs against pathogens commonly encountered in bone infections. PMID- 19307357 TI - Unusual class 1 integron-associated gene cassette configuration found in IncA/C plasmids from Salmonella enterica. AB - IncA/C plasmids carrying an unusual cassette configuration in a class 1 integron and five further shared resistance genes, aacC4, aphA1, hph, sul2, and tetA(D) were found in Salmonella enterica serovars Senftenberg and Ohio. A deletion formed using a short region of homology in the 5' conserved segment and the orfF cassette created an array with only part of orfF followed by the aadA2 cassette. The IncA/C plasmids were not recoverable by conjugation, but additional conjugative resistance plasmids were present in some strains. PMID- 19307358 TI - Preclinical characterization of PF-00868554, a potent nonnucleoside inhibitor of the hepatitis C virus RNA-dependent RNA polymerase. AB - PF-00868554 is a nonnucleoside inhibitor of the hepatitis C virus (HCV) RNA polymerase, which exerts its inhibitory effect by binding to the thumb base domain of the protein. It is a potent and selective inhibitor, with a mean 50% inhibitory concentration of 0.019 microM against genotype 1 polymerases and a mean 50% effective concentration (EC(50)) of 0.075 microM against the genotype 1b Con1 replicon. To determine the in vitro antiviral activity of PF-00868554 against various HCV strains, a panel of chimeric replicons was generated, in which polymerase sequences derived from genotype 1a and 1b clinical isolates were cloned into the 1b-Con1 subgenomic reporter replicon. Our results indicate that PF-00868554 has potent in vitro antiviral activity against a majority (95.8%) of genotype 1a and 1b replicons, with an overall mean EC(50) of 0.059 microM. PF 00868554 showed no cytotoxic effect in several human cell lines, up to the highest concentration evaluated (320 microM). Furthermore, the antiviral activity of PF-00868554 was retained in the presence of human serum proteins. An in vitro resistance study of PF-00868554 identified M423T as the predominant resistance mutation, resulting in a 761-fold reduction in susceptibility to PF-00868554 but no change in susceptibility to alpha interferon and a polymerase inhibitor that binds to a different region. PF-00868554 also showed good pharmacokinetic properties in preclinical animal species. Our results demonstrate that PF 00868554 has potent and broad-spectrum antiviral activity against genotype 1 HCV strains, supporting its use as an oral antiviral agent in HCV-infected patients. PMID- 19307359 TI - Omeprazole antagonizes virulence and inflammation in Salmonella enterica-infected RAW264.7 cells. AB - The proton pump inhibitor omeprazole reduced the intracellular replication of Salmonella enterica serovar Typhimurium in RAW264.7 cells without affecting bacterial growth in vitro or the viability of the host cells. The mechanism was bacteriostatic and interfered with replication mediated by the virulence associated SPI2 type III secretion system. The proton pump inhibitor bafilomycin A(1), in contrast, mediated killing of intracellular bacteria and imposed a marked cytotoxicity on RAW264.7 cells. The two compounds also differentially affected the proinflammatory responses of the infected cells. Bafilomycin A(1) enhanced nitric oxide production, whereas omeprazole delayed IkappaB degradation and blocked nitric oxide production and the secretion of proinflammatory cytokines. These results imply that omeprazole can be used to block the virulence factor-mediated intracellular replication of S. Typhimurium, and that its mechanism of growth inhibition is different from that mediated by bafilomycin A(1). PMID- 19307360 TI - Genital tract, cord blood, and amniotic fluid exposures of seven antiretroviral drugs during and after pregnancy in human immunodeficiency virus type 1-infected women. AB - The objective of the study was to measure antiretroviral exposures in four physiological compartments during pregnancy, delivery, and postpartum. This prospective, open-label, longitudinal study collected paired blood plasma (BP) and genital tract (GT) aspirates antepartum, at delivery, and up to 12 weeks postpartum. Antiretroviral cord BP and amniotic fluid concentrations were also measured. Drug concentrations were analyzed by validated high-performance liquid chromatography/UV and liquid chromatography/tandem mass spectrometry methods, with secondary compartment concentrations presented as the percentage of BP. Fourteen women taking lamivudine plus zidovudine and either lopinavir-ritonavir (n = 7), nelfinavir (n = 6), or nevirapine (n = 1) were enrolled; four also received tenofovir. GT penetration relative to BP was highest for the nucleoside reverse transcriptase inhibitors compared to the protease inhibitors and nevirapine. Only antepartum nelfinavir GT penetration was significantly higher than in the second trimester (geometric mean ratio [GMR], 179.3) or third trimester (GMR, 41.9). Compared to nonpregnant historical controls, antepartum GT penetration was significantly lower (P < 0.05) for zidovudine (GMR, 0.25) and lopinavir (GMR, 0.03); postpartum lopinavir GT penetration continued to be significantly lower (GMR, 0.27). Cord BP exposures were highest for lamivudine and tenofovir (> or = 100%), with cord BP levels of the remaining drugs ranging from 49 to 86% of that of the respective BP level. Amniotic exposures for lamivudine, zidovudine, tenofovir, and nelfinavir were > or = 100%, nevirapine exposure was 53%, and lopinavir and ritonavir exposures were < or = 6% that of BP. We conclude that GT, cord BP, and amniotic fluid exposures vary within and between antiretroviral drug classes and biologic sites. Measurement of antiretroviral exposure in maternal genital secretions, cord BP, and amniotic fluid may be needed to identify signals of subtherapeutic or supratherapeutic drug exposure. PMID- 19307361 TI - Resistance plasmid families in Enterobacteriaceae. PMID- 19307362 TI - Identification of the efflux transporter of the fluoroquinolone antibiotic ciprofloxacin in murine macrophages: studies with ciprofloxacin-resistant cells. AB - Ciprofloxacin, the most widely used totally synthetic antibiotic, is subject to active efflux mediated by a MRP-like transporter in wild-type murine J774 macrophages. To identify the transporter among the seven potential Mrps, we used cells made resistant to ciprofloxacin obtained by long-term exposure to increasing drug concentrations (these cells show less ciprofloxacin accumulation and provide a protected niche for ciprofloxacin-sensitive intracellular Listeria monocytogenes). In the present paper, we first show that ciprofloxacin-resistant cells display a faster efflux of ciprofloxacin which is inhibited by gemfibrozil (an unspecific MRP inhibitor). Elacridar, at a concentration known to inhibit P glycoprotein and breast cancer resistance protein (BCRP), only slightly increased ciprofloxacin accumulation, with no difference between resistant and wild-type cells. Analysis at the mRNA (real-time PCR) and protein (Western blotting) levels revealed an overexpression of Mrp2 and Mrp4. Mrp4 transcripts, however, were overwhelmingly predominant (45% [wild-type cells] to 95% [ciprofloxacin-resistant cells] of all Mrp transcripts tested [Mrp1 to Mrp7]). Silencing of Mrp2 and Mrp4 with specific small interfering RNAs showed that only Mrp4 is involved in ciprofloxacin transport in both ciprofloxacin-resistant and wild-type cells. The study therefore identifies Mrp4 as the most likely transporter of ciprofloxacin in murine macrophages but leaves open a possible common upregulation mechanism for both Mrp4 and Mrp2 upon chronic exposure of eukaryotic cells to this widely used antibiotic. PMID- 19307363 TI - Predictive value of pharmacokinetics-adjusted phenotypic susceptibility on response to ritonavir-enhanced protease inhibitors (PIs) in human immunodeficiency virus-infected subjects failing prior PI therapy. AB - The activities of protease inhibitors in vivo may depend on plasma concentrations and viral susceptibility. This nonrandomized, open-label study evaluated the relationship of the inhibitory quotient (IQ [the ratio of drug exposure to viral phenotypic susceptibility]) to the human immunodeficiency virus type 1 (HIV-1) viral load (VL) change for ritonavir-enhanced protease inhibitors (PIs). Subjects on PI-based regimens replaced their PIs with ritonavir-enhanced indinavir (IDV/r) 800/200 mg, fosamprenavir (FPV/r) 700/100 mg, or lopinavir (LPV/r) 400/200 mg twice daily. Pharmacokinetics were assessed at day 14; follow-up lasted 24 weeks. Associations between IQ and VL changes were examined. Fifty-three subjects enrolled, 12 on IDV/r, 33 on FPV/r, and 8 on LPV/r. Median changes (n-fold) (FC) of 50% inhibitory concentrations (IC(50)s) to the study PI were high. Median 2 week VL changes were -0.7, -0.1, and -1.0 log(10) for IDV/r, FPV/r, and LPV/r. With FPV/r, correlations between the IQ and the 2-week change in VL were significant (Spearman's r range, -0.39 to -0.50; P < or = 0.029). The strongest correlation with response to FPV/r was the IC(50) FC (r = 0.57; P = 0.001), which improved when only adherent subjects were included (r = 0.68; P = 0.001). In multivariable analyses of the FPV/r arm that included FC, one measure of the drug concentration, corresponding IQ, baseline VL, and CD4, the FC to FPV was the only significant predictor of VL decline (P < 0.001). In exploratory analyses of all arms, the area under the concentration-time curve IQ was correlated with the week 2 VL change (r = -0.72; P < 0.001). In conclusion, in PI-experienced subjects with highly resistant HIV-1, short-term VL responses to RTV-enhanced FPV/r correlated best with baseline susceptibility. The IQ improved correlation in analyses of all arms where a greater range of virologic responses was observed. PMID- 19307364 TI - Intracellular localization of the ABCC proteins of Leishmania and their role in resistance to antimonials. AB - The ABCC subfamily of proteins is composed of nine members in Leishmania. We report that all of these proteins have an intracellular localization and that the overexpression of at least four members, ABCC3, ABCC4, ABCC5, and ABCC7, can confer resistance to antimonials, the first-line drug against Leishmania. PMID- 19307365 TI - Tetracycline susceptibility testing and resistance genes in isolates of Acinetobacter baumannii-Acinetobacter calcoaceticus complex from a U.S. military hospital. AB - Infections with multidrug-resistant Acinetobacter baumannii-Acinetobacter calcoaceticus complex bacteria complicate the care of U.S. military personnel and civilians worldwide. One hundred thirty-three isolates from 89 patients at our facility during 2006 and 2007 were tested by disk diffusion, Etest, and broth microdilution for susceptibility to tetracycline, doxycycline, minocycline, and tigecycline. Minocycline was the most active in vitro, with 90% of the isolates tested susceptible. Susceptibilities varied significantly with the testing method. The acquired tetracycline resistance genes tetA, tetB, and tetA(39) were present in the isolates. PMID- 19307366 TI - Comparative antipneumococcal activities of sulopenem and other drugs. AB - For 297 penicillin-susceptible, -intermediate, and -resistant pneumococcal strains, the sulopenem MIC(50)s were 0.008, 0.06, and 0.25, respectively, and the sulopenem MIC(90)s were 0.016, 0.25, and 0.5 microg/ml, respectively. The MIC(50)s of amoxicillin for the corresponding strains were 0.03, 0.25, and 2.0 microg/ml, respectively, and the MIC(90)s were 0.03, 1.0, and 8.0 microg/ml, respectively. The combination of amoxicillin and clavulanate gave MICs similar to those obtained with amoxicillin alone. The sulopenem MICs were similar to those of imipenem and meropenem. The MICs of ss-lactams increased with those of penicillin G, and among the quinolones tested, moxifloxacin had the lowest MICs. Additionally, 45 strains of drug-resistant type 19A pneumococci were tested by agar dilution and gave sulopenem MIC(50)s and MIC(90)s of 1.0 and 2.0 microg/ml, respectively. Both sulopenem and amoxicillin (with and without clavulanate) were bactericidal against all 12 strains tested at 2x MIC after 24 h. Thirty-one strains from 10 countries with various penicillin, amoxicillin, and carbapenems MICs, including those with the highest sulopenem MICs, were selected for sequencing analysis of the pbp1a, pbp2x, and pbp2b regions encoding the transpeptidase active site and MurM. We did not find any correlations between specific penicillin-binding protein-MurM patterns and changes in the MICs. PMID- 19307367 TI - Colorimetric high-throughput screen for detection of heme crystallization inhibitors. AB - Malaria infects 500 million people annually, a number that is likely to rise as drug resistance to currently used antimalarials increases. During its intraerythrocytic stage, the causative parasite, Plasmodium falciparum, metabolizes hemoglobin and releases toxic heme, which is neutralized by a parasite-specific crystallization mechanism to form hemozoin. Evidence suggests that chloroquine, the most successful antimalarial agent in history, acts by disrupting the formation of hemozoin. Here we describe the development of a 384 well microtiter plate screen to detect small molecules that can also disrupt heme crystallization. This assay, which is based on a colorimetric assay developed by Ncokazi and Egan (K. K. Ncokazi and T. J. Egan, Anal. Biochem. 338:306-319, 2005), requires no parasites or parasite-derived reagents and no radioactive materials and is suitable for a high-throughput screening platform. The assay's reproducibility and large dynamic range are reflected by a Z factor of 0.74. A pilot screen of 16,000 small molecules belonging to diverse structural classes was conducted. The results of the target-based assay were compared with a whole parasite viability assay of the same small molecules to identify small molecules active in both assays. PMID- 19307368 TI - Combination therapy in treatment of experimental pulmonary aspergillosis: in vitro and in vivo correlations of the concentration- and dose- dependent interactions between anidulafungin and voriconazole by Bliss independence drug interaction analysis. AB - We studied the antifungal activity of anidulafungin (AFG) in combination with voriconazole (VRC) against experimental invasive pulmonary aspergillosis (IPA) in persistently neutropenic rabbits and further explored the in vitro and in vivo correlations by using Bliss independence drug interaction analysis. Treatment groups consisted of those receiving AFG at 5 (AFG5 group) and 10 (AFG10 group) mg/kg of body weight/day, VRC at 10 mg/kg every 8 h (VRC group), AFG5 plus VRC (AFG5+VRC group), and AFG10 plus VRC (AFG10+VRC group) and untreated controls. Survival throughout the study was 60% for the AFG5+VRC group, 50% for the VRC group, 27% for the AFG10+VRC group, 22% for the AFG5 group, 18% for the AFG10 group, and 0% for control rabbits (P < 0.001). There was a significant reduction of organism-mediated pulmonary injury, measured by infarct scores, lung weights, residual fungal burdens, and galactomannan indexes, in AFG5+VRC-treated rabbits versus those treated with AFG5 and VRC alone (P < 0.05). In comparison, AFG10+VRC significantly lowered only infarct scores and lung weights in comparison to those of AFG10-treated animals (P < 0.05). AFG10+VRC showed no significant difference in other outcome variables. Significant Bliss synergy was found in vivo between AFG5 and VRC, with observed effects being 24 to 30% higher than expected levels if the drugs were acting independently. These synergistic interactions were also found between AFG and VRC in vitro. However, for AFG10+VRC, only independence and antagonism were observed among the outcome variables. We concluded that the combination of AFG with VRC in treatment of experimental IPA in persistently neutropenic rabbits was independent to synergistic at a dosage of 5 mg/kg/day but independent to antagonistic at 10 mg/kg/day, as assessed by Bliss independence analysis, suggesting that higher dosages of an echinocandin may be deleterious to the combination. PMID- 19307369 TI - Atorvastatin is a promising partner for antimalarial drugs in treatment of Plasmodium falciparum malaria. AB - Atorvastatin (AVA) is a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor. AVA exposure resulted in the reduced in vitro growth of 22 Plasmodium falciparum strains, with the 50% inhibitory concentrations (IC(50)s) ranging from 2.5 microM to 10.8 microM. A significant positive correlation was found between the strains' responses to AVA and mefloquine (r = 0.553; P = 0.008). We found no correlation between the responses to AVA and to chloroquine, quinine, monodesethylamodiaquine, lumefantrine, dihydroartemisinin, atovaquone, or doxycycline. These data could suggest that the mechanism of AVA uptake and/or the mode of action of AVA is different from those for other antimalarial drugs. The IC(50)s for AVA were unrelated to the occurrence of mutations in the transport protein genes involved in quinoline antimalarial drug resistance, such as the P. falciparum crt, mdr1, mrp, and nhe-1 genes. Therefore, AVA can be ruled out as a substrate for the transport proteins (CRT, Pgh1, and MRP) and is not subject to the pH modification induced by the P. falciparum NHE-1 protein. The absence of in vitro cross-resistance between AVA and chloroquine, quinine, mefloquine, monodesethylamodiaquine, lumefantrine, dihydroartemisinin, atovaquone, and doxycycline argues that these antimalarial drugs could potentially be paired with AVA as a treatment for malaria. In conclusion, the present observations suggest that AVA is a good candidate for further studies on the use of statins in association with drugs known to have activities against the malaria parasite. PMID- 19307370 TI - Antifungal activities of natural and synthetic iron chelators alone and in combination with azole and polyene antibiotics against Aspergillus fumigatus. AB - Antifungal effects of iron chelators (lactoferrin, deferoxamine, deferiprone, and ciclopirox) were tested alone and in combination with antifungal drugs against Aspergillus fumigatus B5233 conidia. Lactoferrin, ciclopirox, and deferiprone inhibited whereas deferoxamine enhanced fungal growth. Antifungal synergy against conidia was observed for combinations of ketoconazole with ciclopirox or deferiprone, lactoferrin with amphotericin B, and fluconazole with deferiprone. Iron chelation alone or combined with antifungal drugs may be useful for prevention and treatment of mycosis. PMID- 19307371 TI - In vitro activity of micafungin against planktonic and sessile Candida albicans isolates. AB - Planktonic and sessile susceptibilities to micafungin were determined for 30 clinical isolates of Candida albicans obtained from blood or other sterile sites. Planktonic and sessile MIC(90)s for micafungin were 0.125 and 1.0 microg/ml, respectively. PMID- 19307372 TI - Antifungal combinations against simulated Candida albicans endocardial vegetations. AB - The in vitro effects of flucytosine (5FC), liposomal amphotericin B (L-AmB), and micafungin (Mica) combinations against two Candida albicans strains that simulated 24-hour-old endocardial vegetations were studied. Mica was superior to 5FC or L-AmB, and the 5FC-L-AmB-Mica combination was superior to all other treatments for one strain but no different from the dual combination of L-AmB Mica for the other strain. PMID- 19307373 TI - In vivo selection of reduced susceptibility to carbapenems in Acinetobacter baumannii related to ISAba1-mediated overexpression of the natural bla(OXA-66) oxacillinase gene. AB - Two clonally related Acinetobacter baumannii isolates, A1 and A2, were obtained from the same patient. Isolate A2, selected after an imipenem-containing treatment, showed reduced susceptibility to carbapenems. This resistance pattern was related to insertion of the ISAba1 element upstream of the naturally occurring bla(OXA-66) carbapenemase gene as demonstrated by sequencing, reverse transcription-PCR analysis, and inactivation of the bla(OXA-66) gene. PMID- 19307374 TI - P1A recombinant beta-lactamase prevents emergence of antimicrobial resistance in gut microflora of healthy subjects during intravenous administration of ampicillin. AB - Ipsat P1A is a recombinant beta-lactamase which degrades antibiotic residue in the gastrointestinal tract. In an open-label, single-center controlled trial, 36 healthy subjects were randomized to receive (i) ampicillin (1 g intravenously [i.v.] every 6 h [q6h]), (ii) oral P1A recombinant beta-lactamase (8.2 mg q6h), or (iii) ampicillin (1 g i.v. q6h) in combination with oral P1A recombinant beta lactamase (8.2 mg q6h) for 5 days. Fecal samples were collected before treatment, during treatment (days 3 to 5), and at follow-up (day 12). The primary end points were (i) changes in gastrointestinal microflora (determined by temperature gradient gel electrophoresis [TGGE]) and (ii) emergence of bacterial resistance (determined by conventional microbiology and PCR of TEM beta-lactamase genes). Thirty-five subjects completed the study. The mean similarity percentages of TGGE profiles between baseline and each treatment day sample were significantly lower for the ampicillin group than for the group receiving ampicillin plus P1A recombinant beta-lactamase on days 3, 4, and 5 (P < 0.001). Compared with the ampicillin group, subjects receiving ampicillin plus P1A recombinant beta lactamase had significantly fewer ampicillin-resistant coliforms on days 3, 4, and 5 and at follow-up (P < or = 0.001) and fewer TEM beta-lactamase genes on days 3, 4, and 5 (P < 0.02). P1A recombinant beta-lactamase was safe and well tolerated. In healthy subjects, P1A recombinant beta-lactamase prevents ampicillin-induced alterations in intestinal microflora, emergence of resistance, and the number of TEM genes. PMID- 19307375 TI - The chromosomal toxin gene yafQ is a determinant of multidrug tolerance for Escherichia coli growing in a biofilm. AB - Escherichia coli is refractory to elevated doses of antibiotics when it is growing in a biofilm, and this is potentially due to high numbers of multidrug tolerant persister cells in the surface-adherent population. Previously, the chromosomal toxin-antitoxin loci hipBA and relBE have been linked to the frequency at which persister cells occur in E. coli populations. In the present study, we focused on the dinJ-yafQ-encoded toxin-antitoxin system and hypothesized that deletion of the toxin gene yafQ might influence cell survival in antibiotic-exposed biofilms. By using confocal laser scanning microscopy and viable cell counting, it was determined that a Delta yafQ mutant produced biofilms with a structure and a cell density equivalent to those of the parental strain. In-depth susceptibility testing identified that relative to wild-type E. coli, the Delta yafQ strain had up to a approximately 2,400-fold decrease in cell survival after the biofilms were exposed to bactericidal concentrations of cefazolin or tobramycin. Corresponding to these data, controlled overexpression of yafQ from a high-copy-number plasmid resulted in up to a approximately 10,000 fold increase in the number of biofilm cells surviving exposure to these bactericidal drugs. In contrast, neither the inactivation nor the overexpression of yafQ affected the tolerance of biofilms to doxycycline or rifampin (rifampicin). Furthermore, deletion of yafQ did not affect the tolerance of stationary-phase planktonic cells to any of the antibacterials tested. These results suggest that yafQ mediates the tolerance of E. coli biofilms to multiple but specific antibiotics; moreover, our data imply that this cellular pathway for persistence is likely different from that of multidrug-tolerant cells in stationary-phase planktonic cell cultures. PMID- 19307376 TI - Inhibition of vaccinia virus replication by two small interfering RNAs targeting B1R and G7L genes and their synergistic combination with cidofovir. AB - In view of the threat of the potential use of variola virus in a terrorist attack, considerable efforts have been performed to develop new antiviral strategies against orthopoxviruses. Here we report on the use of RNA interference, either alone or in combination with cidofovir, as an approach to inhibit orthopoxvirus replication. Two selected small interfering RNAs (siRNAs), named siB1R-2 and siG7L-1, and a previously reported siRNA, i.e., siD5R-2 (which targets the viral D5R mRNA), were evaluated for antiviral activity against vaccinia virus (VACV) by plaque reduction and virus yield assays. siB1R-2 and siG7L-1, administered before or after viral infection, reduced VACV replication by more than 90%. Also, these two siRNAs decreased monkeypox virus replication by 95% at a concentration of 1 nM. siB1R-2 and siG7L-1 were demonstrated to specifically silence their corresponding transcripts, i.e., B1R and G7L mRNAs, without induction of a beta interferon response. Strong synergistic effects were observed when siB1R-2, siG7L-1, or siD5R-2 was combined with cidofovir. In addition, the antiviral activities of these three siRNAs were evaluated against VACV resistant to cidofovir and other acyclic nucleoside phosphonates. siG7L-1 and siD5R-2 remained active against four of five VACV mutants, while siB1R-2 showed activity against only one of the mutants. Our results showed that siRNAs are potent inhibitory agents in vitro, not only against wild-type VACV but also against several cidofovir-resistant VACV. Furthermore, we showed that a combined therapy using siRNA and cidofovir may be useful in the treatment of poxvirus infections. PMID- 19307377 TI - Evaluation of cefotaxime and desacetylcefotaxime concentrations in cord blood after intrapartum prophylaxis with cefotaxime. AB - Preterm premature rupture of the membranes is associated with a high risk of neonatal sepsis. An increase in the incidence of early-onset neonatal sepsis due to ampicillin-resistant Escherichia coli in premature infants has been observed in the past few years. Intrapartum prophylaxis with ampicillin has proven to be efficient for the prevention of early neonatal sepsis due to group B streptococci. To date, there is no strategy for the prevention of early neonatal sepsis due to ampicillin-resistant E. coli. Our aim was to investigate whether a standardized dosage regimen of intrapartum cefotaxime could provide concentrations in the cord blood greater than the cefotaxime MIC(90) for E. coli. Seven pregnant women hospitalized with preterm premature rupture of the membranes and colonized with ampicillin-resistant isolates of the family Enterobacteriaceae were included. Cefotaxime was given intravenously during delivery, as follows: 2 g at the onset of labor and then 1 g every 4 h until delivery. Blood specimens were collected from the mother 30 min after the first injection and just before the second injection, and at birth, blood specimens were simultaneously collected from the mother and the umbilical cord. The concentrations of cefotaxime in the cord blood ranged from 0.5 to 8.5 mg/liter. The MIC(90) of cefotaxime for E. coli strains (0.125 mg/liter) was achieved in all cases. This preliminary study supports the use of cefotaxime for intrapartum prophylaxis in women colonized with ampicillin-resistant isolates of Enterobacteriaceae. The effectiveness of this regimen for the prevention of neonatal sepsis needs to be evaluated with a larger population. PMID- 19307378 TI - Children's injuries: a global problem. PMID- 19307380 TI - On the philosophy of diagnosis: is doing more good than harm better than "primum non nocere"? AB - Diagnosis is arguably the cornerstone of medicine. Without at least some form of diagnosis the practice of medicine would not be possible. This narrative review explores common philosophical assumptions and challenges the notion that a certain diagnosis can ever be made. The idealistic concept of "primum non nocere" is discussed, and whether the utilitarian goal of achieving "the greatest happiness for the greatest number" is a feasible or preferable alternative is considered. It is concluded that utilitarianism is inescapably intertwined with modern medical practice. Suggestions are presented to further the understanding of diagnostic medicine by embracing its principles. PMID- 19307381 TI - Predicting patient arrivals to an accident and emergency department. AB - OBJECTIVES: To characterise and forecast daily patient arrivals into an accident and emergency (A&E) department based on previous arrivals data. METHODS: Arrivals between 1 April 2002 and 31 March 2007 to a busy case study A&E department were allocated to one of two arrival streams (walk-in or ambulance) by mode of arrival and then aggregated by day. Using the first 4 years of patient arrival data as a "training" set, a structural time series (ST) model was fitted to characterise each arrival stream. These models were used to forecast walk-in and ambulance arrivals for 1-7 days ahead and then compared with the observed arrivals given by the remaining 1 year of "unseen" data. RESULTS: Walk-in arrivals exhibited a strong 7-day (weekly) seasonality, with ambulance arrivals showing a distinct but much weaker 7-day seasonality. The model forecasts for walk-in arrivals showed reasonable predictive power (r = 0.6205). However, the ambulance arrivals were harder to characterise (r = 0.2951). CONCLUSIONS: The two separate arrival streams exhibit different statistical characteristics and so require separate time series models. It was only possible to accurately characterise and forecast walk-in arrivals; however, these model forecasts will still assist hospital managers at the case study hospital to best use the resources available and anticipate periods of high demand since walk-in arrivals account for the majority of arrivals into the A&E department. PMID- 19307382 TI - Resuscitation tables: a useful tool in calculating pre-burns unit fluid requirements. AB - BACKGROUND: There is considerable variation in the standard of initial burn management, particularly burn surface area assessment and application of resuscitation formulae. Early aggressive management of major burns improves survival. Internationally, the Parkland formula employing lactated Ringer's solution is used for fluid resuscitation. This study aimed to assess whether Parkland fluid resuscitation tables could improve the accuracy of initial fluid requirement calculations. METHODS: The burn size had first to be determined for an adult and a child using a preshaded Lund and Browder chart. Fluid requirements then had to be calculated using the conventional Parkland formula. The burn size had to be similarly calculated for two further cases and fluid requirements calculated using resuscitation tables. The study had a sample size of 50, consisting of plastic surgery trainees, anaesthetists and burn nurse specialists. RESULTS: All the participants found the resuscitation tables to be quicker and easier to use. The burn size was correctly calculated in 72% of cases. Fluid resuscitation requirements were correct in only 55% when using the Parkland formula. The use of resuscitation tables improved the accuracy in calculating fluid requirements to 75%. CONCLUSIONS: The use of Parkland fluid resuscitation tables can improve accuracy and ease of calculation of fluid resuscitation requirements. PMID- 19307383 TI - Can the Ottawa knee rule be applied to children? A systematic review and meta analysis of observational studies. AB - BACKGROUND: The Ottawa knee rule (OKR), a clinical decision aid is used to reduce unnecessary radiography. It is not clear whether this rule can be applied to children. OBJECTIVE: To establish whether the OKR had adequate sensitivity and acceptable specificity in children to advocate widespread use. METHODS: A systematic review and meta-analysis was conducted of observational studies that examined the diagnostic characteristics of the OKR in children. DATA SOURCES: Relevant English language articles were identified from Medline (1950 to date), EMBASE (1974 to date), CINAHL (1982 to date), the Cochrane Library, Google Scholar and a hand search of bibliographies. STUDY SELECTION: Observational studies that included children and have used the OKR for ruling out fractures in children either radiologically or in combination with follow-up. RESULTS: Four relevant studies were identified. Three studies were suitable for inclusion in the meta-analysis, representing 1130 children. The pooled negative likelihood ratio was 0.07 (95% CI 0.02 to 0.29), the pooled positive likelihood ratio was 1.94 (95% CI 1.60 to 2.36), the pooled sensitivity was 99% (CI 94.4 to 99.8) and the pooled specificity was 46% (CI 43.0 to 49.1). The reduction in radiography was between 30% and 40%. CONCLUSION: The OKR has high sensitivity and adequate specificity for children over the age of 5 years. There are not enough good data to advocate application of the OKR in children less than 5 years. PMID- 19307384 TI - Prediction of mortality in adult emergency department patients with sepsis. AB - OBJECTIVES: To determine the efficacy of the abbreviated Mortality in Emergency Department Sepsis (MEDS) score, the Modified Early Warning (MEW) score and near patient-test (NPT) lactate levels in predicting 28-day mortality in adult emergency department (ED) patients with sepsis. METHODS: A retrospective cohort study of adult ED patients with sepsis admitted to hospital was conducted in a large urban teaching and a district general hospital. Data were collected during four time periods between 1 January 2006 and 31 January 2007. Inclusion criteria were age > or =16 years and an ED diagnosis of sepsis. Primary outcome for all patients was 28-day mortality. Patients were preassigned to risk groups according to their abbreviated MEDS score, MEW score and NPT lactate. RESULTS: 307 ED patients with sepsis were included in the study. Among these there were 72 deaths (23%). Mortality rates for the low-, moderate- and high-risk groups of the abbreviated MEDS score were 1/63 (1.6%), 48/205 (23.4%) and 23/39 (59.0%) patients. The MEDS score for low-risk patients was 98.6% (95% CI 92.5% to 99.9%) sensitive and 26.5% (95% CI 21.0% to 32.6%) specific and for high-risk patients it was 31.9% (95% CI 21.4% to 44.0%) sensitive and 93.2% (95% CI 89.2% to 96.1%) specific for death within 28 days. Mortality rates for the low- and high-risk MEW score were 20/159 (12.6%) and 52/148 (35.1%) patients. The MEW score for high risk patients was 72.2% (95% CI 60.4% to 82.1%) sensitive and 59.2% (95% CI 52.6% to 65.5%) specific for mortality. An NPT lactate level of > or =4 mmol/l was 49.1% (95% CI 35.1% to 63.2%) sensitive and 74.3% (95% CI 64.8% to 82.3%) specific for 28-day mortality. CONCLUSION: These results demonstrate the efficacy of the abbreviated MEDS score, the MEW score and NPT venous lactate levels in predicting 28-day mortality in ED patients with sepsis. The abbreviated MEDS score was found to be the best performing risk assessment model which, with prospective validation, may aid early clinical decision-making in ED patients with sepsis and might affect the outcome from sepsis. PMID- 19307385 TI - Patients from care homes who attend the emergency department: could they be managed differently. AB - AIMS: To describe the demographics of patients from care homes who attend the emergency department (ED) and to estimate whether appropriate alternative care pathways could have been delivered to this patient group. METHODS: A prospective descriptive study that gathered data on consecutive patients who presented to the ED of the Royal Infirmary of Edinburgh from adult care homes over a one-month period. A proforma was used to record details such as diagnosis, communication from the care home, referral details, patient transport and resuscitation status. Three general practitioners (GPs) assessed whether the patients could have been managed appropriately outwith the ED. RESULTS: 114 patients were recruited over the month of the study. Seven patients were missed. Results showed that 40 care home patients (35%) came to the ED without an accompanying letter. 99 patients (87%) were brought to the department by ambulance. 58 patients (51%) who came to the ED from care homes were discharged, and of these 41 (71%) were transported back to the care home by non-ambulance transport. Three patients (3%) had a resuscitation status documented. GP review of case notes suggested that between nine (8%) and 46 (40%) could have been managed appropriately outwith the ED. CONCLUSION: In this study between 8% and 40% of patients could have been cared for outwith the ED. The findings highlight the importance of the provision of appropriate healthcare to care home residents and the current deficits that exist. PMID- 19307386 TI - Characteristics of frequent attenders in an emergency department: analysis of 1 year attendance data. AB - BACKGROUND: There is a significant literature examining the reasons why patients attend emergency departments frequently. This body of work suggests that sociodemographic characteristics are important in understanding why patients re attend, but it does not provide a definition of what frequent attendance means. This paper aims to identify personal and attendance factors associated with frequent attendance at an emergency department. METHODS: One-year emergency department attendance data from a south-east London teaching hospital (2006-7) were analysed. The dataset was analysed at two levels: the individual patient level and the attendance level. Frequencies and cross-tabulations were produced to describe the dataset. Confidence intervals were calculated for both patient and attendance level data. RESULTS: 82 812 patients made 117 187 attendances to the emergency department during 1 year. Each patient made an average of 1.4 attendances; 46% were repeat attendances. The analysis demonstrated differences in the personal and attendance profile of patients who attended the emergency department more frequently during the study period. A change in the patient profile first appeared at the fourth attendance and the change became more pronounced as attendances increased. Frequent attenders were more likely to be men (50.5% of single attendances; 69.5% of > or =10 attendances), older (single attendance, mean age 32 years; > or =10 attendances, mean age 45.6 years), to attend outside daytime hours (51.4% of single attendances; 69.2% of > or =10 attendances) and to be triaged into the more serious categories (36.1% of single attendances; 54.3% of > or =10 attendances). CONCLUSION: Where local services are being designed to divert frequent attenders, existing data sources can be a rich source of information to inform service design. For example, this analysis identifies older men at their fourth or more attendance as a potentially important group when examining frequent attendance at this particular hospital. It also identified a potential need for services outside normal surgery hours, although frequent attenders tend to be triaged into the more urgent categories. PMID- 19307387 TI - Evaluation of a method for converting venous values of acid-base and oxygenation status to arterial values. AB - OBJECTIVE: This paper evaluates a method in which arterial values of pH, carbon dioxide tension (Pco(2)) and oxygen tension (Po(2)) calculated from venous values and pulse oximetry are compared with simultaneously measured arterial values. METHODS: 103 adult patients from three departments (pulmonary medicine, thoracic intensive care and multidisciplinary intensive care) were studied. The patients belonged to three groups: (1) 31 haemodynamically stable patients with a diagnosis of chronic obstructive lung disease (COLD); (2) 49 haemodynamically stable patients without COLD; and (3) 23 haemodynamically unstable patients without COLD. Arterial and venous (peripheral and, where possible, central and mixed) blood samples were taken simultaneously and anaerobically. Peripheral arterial oxygen saturation was measured with a pulse oximeter. The principle of the method is to simulate the transport of venous blood back through the tissues using the respiratory quotient (adding oxygen and removing carbon dioxide) until simulated arterial oxygenation matches that measured by pulse oximetry. RESULTS: Calculated values of arterial pH and Pco(2) had very small bias and standard deviations regardless of the venous sampling site. In all cases these errors were within those considered acceptable for the performance of laboratory equipment, and well within the limits of error acceptable in clinical practice. In addition, the standard deviation (SD) of calculated values of pH and Pco(2) was similar to the variability between consecutive arterial samples. For peripheral oxygen saturation values < or =96%, the method can calculate Po(2) with an SD of 0.93, which may be useful in clinical practice. Calculations made from peripheral venous blood were significantly more accurate than those from central venous blood. CONCLUSION: Arterial pH and Pco(2) can be calculated precisely from peripheral venous blood in a broad patient population. The method has potential for use as a screening tool in emergency medical departments and in medical and surgical wards to assess a patient's acid-base and oxygenation status prior to sampling arterial blood or to help in the decision to refer the patient to the ICU. In departments where arterial blood gas values are used to monitor patients (eg, pulmonary medicine), the method might reduce the number of arterial samples taken by replacing them with peripheral venous blood samples, thus reducing the need for painful arterial punctures. PMID- 19307388 TI - Diagnosis of pyogenic liver abscess by abdominal ultrasonography in the emergency department. AB - BACKGROUND: Delayed diagnosis of pyogenic liver abscess remains a challenging problem in the emergency department because of the associated high morbidity and mortality. OBJECTIVE: To evaluate the sensitivity of ultrasono-graphy in the diagnosis of pyogenic liver abscess in patients presenting to the emergency department and the factors that may influence this sensitivity. METHODS: A retrospective study was conducted in patients diagnosed with pyogenic liver abscess in the emergency department (ED) of a tertiary care teaching hospital for a period of 5 years. Between May 2001 and April 2006, 268 patients diagnosed with pyogenic liver abscess were evaluated by ultrasonography and/or CT scanning. The age, sex, clinical presentation, location and number of abscesses and the underlying disease of these two groups were compared. RESULTS: Of the 268 patients admitted via the ED who were discharged or died with a diagnosis of pyogenic liver abscess, there was a predominance of men (M/F 173/95) and the mean age was 57.6 years (range 17-90). 38 had false negative findings on ultrasonography (sensitivity 85.8%) and required abdominal CT scanning for definitive diagnosis. In the other 230 cases, ultrasonography alone was sufficient for diagnosis. Location of the abscess in segments 4 and 5 of the liver raised the sensitivity of ultrasound for diagnosis, while location in segment 8 was most associated with delayed diagnosis by ultrasonography. Right costal angle knocking pain was significant for pyogenic liver abscess even if ultrasound was negative. CONCLUSIONS: The size and location of the liver abscess and the underlying comorbid diseases may affect the diagnostic sensitivity of ultrasound for pyogenic liver abscess in clinical practice. A high index of suspicion should be maintained in patients with diabetes mellitus, previous biliary tract intervention or gastrointestinal malignancy. Follow-up CT scanning is recommended if right flank knocking pain is present, even if ultrasonography is non-revealing. A diagnostic protocol for liver abscess may be feasible in the future. PMID- 19307389 TI - Ultrasound identification of landmarks preceding lumbar puncture: a pilot study. AB - AIM: To assess the utility of bedside ultrasound performed by an emergency physician in adults undergoing diagnostic lumbar puncture. METHOD: Ultrasound was used as the primary means of determining the site of skin puncture, angle of needle advancement and depth needed to access the subarachnoid space. RESULTS: Cerebrospinal fluid was obtained from 36 of 39 patients (92.3%) in the first interspinous space attempted. CONCLUSIONS: The ultrasonographically measured depth of the dura mater correlates strongly with the final needle depth. PMID- 19307390 TI - Feasibility of a structured 3-minute walk test as a clinical decision tool for patients presenting to the emergency department with acute dyspnoea. AB - INTRODUCTION: Emergency department (ED) physicians face frequent decisions on whether to admit patients with congestive heart failure (CHF) or acute exacerbation of chronic obstructive pulmonary disease (COPD). This feasibility study evaluated the potential of a structured 3-minute walk test as a clinical decision tool for admission and correlated its performance with poor clinical outcomes. It also aimed to gather evidence and directions for the design of a multicentre study to derive clinical guidelines. METHODS: In this prospective cohort study, a convenience sample was enrolled of 40 adult patients who presented to a tertiary care ED with CHF, COPD, or stable chest pain and were being considered for discharge. Patients walked at their own pace and their dyspnoea, respiratory rate, heart rate and oxygen saturation were recorded each minute for 4 minutes. The primary outcome was "poor clinical outcome" defined as admission to hospital, the need for biphasic positive airway pressure, the need for intubation, relapse, or death. RESULTS: 85.0% successfully completed the test and 30.0% had poor clinical outcomes. Of those with poor clinical outcomes, 41.7% were unable to complete the test compared with only 3.6% of those with good clinical outcomes (p<0.01). Significant differences were noted in the dyspnoea (p = 0.04) and respiratory rate (p = 0.03) as well as oxygen saturation measurements at 3 minutes. CONCLUSIONS: The 3-minute walk test is a non-resource intensive, simple procedure with applicability in most ED for discharge decisions in patients with cardiopulmonary conditions. Multicentre studies are being planned to validate these findings and establish guidelines for admission and discharge of patients with CHF or acute exacerbation of COPD. PMID- 19307391 TI - Westermark's sign in submassive pulmonary embolism. PMID- 19307392 TI - Blunt abdominal trauma in children: how predictive is ALT for liver injury? AB - BACKGROUND: The liver is the second most commonly injured intra-abdominal organ in children. CT scanning is currently regarded as the "gold standard" in screening for intra-abdominal injury following blunt trauma. However, the risks associated with performing CT in children are not insignificant and, in addition, CT is not always readily available. This study investigates the utility of alanine aminotransferase (ALT) in screening for liver injury in paediatric trauma. METHODS: Two groups of patients were compiled from a prospectively identified trauma registry-one with liver injuries and another with intra abdominal injuries other than to the liver. Inclusion in the study required that an initial ALT level had been obtained after injury. Where CT had been performed, a paediatric radiologist blind to the ALT results graded the severity of the liver injuries. The study groups were compared and a receiver operating characteristic (ROC) curve generated to derive the optimum ALT threshold to identify liver injury. RESULTS: 51 patients with liver injury and 65 with other intra-abdominal injuries were identified. An ALT level of > or =104 IU/l gave a sensitivity of 96% and a specificity of 80%. When liver injuries were stratified to identify only clinically significant liver injuries (grades III, IV and V), this ALT threshold identified 100% of patients with 70% specificity. CONCLUSIONS: In this sample, ALT appears to be a useful predictor for the presence or absence of liver injury. In haemodynamically stable children with clinical suspicion of isolated liver injury, identification of a normal ALT level (<104 IU/l) may reduce the need for unnecessary transportation for CT scanning with subsequent radiation exposure. PMID- 19307393 TI - Right person? Right place? PMID- 19307394 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. BET 2. Diagnostic utility of chest X ray for investigation of pulmonary embolism. PMID- 19307395 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. BET 1. Flight deep vein thrombosis prophylaxis in lower limb injury. PMID- 19307397 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. BET3. Normal partial pressure oxygen in room air as a rule-out for pulmonary embolism. PMID- 19307398 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. BET 4. Dose of dexamethasone in croup. PMID- 19307399 TI - A single, improvised "Kassam" rocket explosion can cause a mass casualty incident: a potential threat for future international terrorism? AB - INTRODUCTION: Over 2000 improvised rockets (called "Kassam" rockets) have been targeted at the south of Israel from the Gaza strip since 2001. Most of them have injured relatively few people. The first known case of a multicasualty incident (MCI) caused by the landing of a single, improvised rocket is described. METHODS: The event is described according to the disastrous incidents systematic analysis through components, interactions and results methodology (DISAST-CIR). RESULTS: The rocket hit a military training tent camp in the south of Israel at 01:18 hours. At that time, all soldiers were in bed and were not using any protective gear. A total of 76 soldiers was injured (three severe, eight moderate and 65 mild). The most prevalent types of injuries were upper extremity (33%) and lower extremity (30%) trauma, tinnitus (30%) and acute stress reactions (32%). A total of 67 casualties was evacuated to the nearest level two hospital, Barzilai, in a two-phase distribution characterised by different patterns of injury severity and type. All urgent casualties arrived at hospitals within 1 h 24 minutes, whereas most stress casualties arrived in the later phase. Seven casualties were secondarily transported to level one trauma centres. 42 of the casualties were hospitalised and 17 needed urgent surgery. None has died. CONCLUSIONS: A single low-tech mortar with poor accuracy and small warhead (estimated weight of 10 kg only) can cause a large-scale MCI. As international terrorist organisations can easily gain access to improvised rockets, the latter may become a threat in many countries. Emergency systems should thus be prepared for that adverse possibility. PMID- 19307401 TI - The Mount Fuji sign of the pneumoperitoneum. PMID- 19307400 TI - Acute pulmonary embolism presenting as seizures. AB - Pulmonary embolism (PE) is a common and potentially fatal condition. The case is presented of a young woman who presented to the emergency department with first time seizures. On further investigation and corroborated by post-mortem findings, this was found to be a manifestation of PE. The purpose of this report is to highlight to the clinical community that PE can present in a varied fashion and that, because of this, its diagnosis can occasionally be delayed. Vigilance is advised and expert clinical acumen is needed in young non-smoking persons with previously normal respiratory physiology who present with signs and symptoms of respiratory compromise, irrespective of the predominant or masking presentation. PMID- 19307402 TI - A case of ricin poisoning following ingestion of Korean castor bean. AB - Ricin is a member of a group of protein toxins whose cytosolic target is the 28S rRNA of the 60S ribosomal subunit. It was recently introduced as a weapon of terrorism in some countries. There is little evidence about the mechanism, diagnosis and treatment of ricin poisoning. The case history is reported of a patient with ricin poisoning who presented with nausea and vomiting following ingestion of Korean castor bean. PMID- 19307403 TI - Penetration of the iliac wing by retained foreign body. PMID- 19307404 TI - Caveats in the management and diagnosis of cerebellar infarct and vertebral artery dissection. AB - Early MRI, available 24 h a day and at weekends, will make a timely diagnosis in cases of cerebellar infarct and a normal initial CT scan does not exclude it. In many emergency departments MRI is not as routinely available out of hours as CT. It is important to appreciate the varied symptomatology and signs of cerebellar infarcts in order to avoid misdiagnosis or delayed diagnosis and to remember that, apart from requiring treatment with anticoagulation for the cerebellar infarct itself, the complications of cerebellar infarct may require surgical intervention. Vertebral artery dissection as a cause of cerebellar infarct may require anticoagulation or endovascular therapy. PMID- 19307405 TI - Retinal detachment diagnosed by magnetic resonance imaging. PMID- 19307406 TI - An unusual cause of ST elevation. AB - An interesting case of a patient presenting to the emergency department with chest pain and an electrocardiogram (ECG) revealing ST elevation is presented. On closer examination another explanation for this ECG appearance is discussed indicating that emergency revascularisation was not indicated. PMID- 19307407 TI - Pseudarthrosis of the tibia: emergency department presentation of neurofibromatosis type 1 in a 4-month-old infant. AB - Congenital pseudarthrosis of the tibia is a complex and rare condition of infancy. Long bone pseudarthrosis is strongly linked to neurofibromatosis, a common genetic disorder in which pseudarthrosis tends to occur early. This report describes a similar emergency department presentation in a 4-month-old infant with subsequent diagnosis of neurofibromatosis type 1. PMID- 19307409 TI - Potter's potions: aconite poisoning. PMID- 19307410 TI - Surveillance of poisons centre enquiries. PMID- 19307411 TI - Clinical feedback to ambulance crews: supporting professional development. PMID- 19307412 TI - A lost message: why are doctors so resistant to change? PMID- 19307413 TI - Emergency department testing for Streptococcus in children with sore throats. PMID- 19307414 TI - Postpartum emergencies. PMID- 19307415 TI - Advanced trauma life support: back to basics. PMID- 19307417 TI - Reproductive physiology: songbird study removes long-standing neuroendocrinology research roadblock. PMID- 19307418 TI - G protein-coupled receptor 30: estrogen receptor or collaborator? PMID- 19307419 TI - Prolactin receptors are critical to the adaptation of islets to pregnancy. PMID- 19307420 TI - Ischemic injury to white matter: an age-dependent process. AB - The risk for ischemic stroke increases drastically with age, although reasons for this remain unexplored. White matter (WM) and gray matter constitute equal proportions of the brain, and WM is injured in most strokes. Axonal injury and dysfunction are responsible for much of the disability associated with clinical deficits observed after stroke. The authors recently reported that central nervous system WM is inherently more vulnerable to ischemic injury in older mice, and the mechanisms of WM injury change as a function of age. Ischemic WM injury in older mice is predominantly mediated by a Ca2+-independent excitotoxicity involving overactivation of AMPA/kainate receptors. Glutamate release, due to reverse glutamate transport, occurs earlier and is more robust in older mice that show up-regulation of GLT1, the main glutamate transporter. Blockade of NMDA receptors does not improve WM function after ischemia in the young but aggravates ischemic injury in older mice. The main goals of this research update are to summarize the evidence for equivalent brain insults inducing more damage with aging, and to highlight the importance of age in any successful stroke therapy. PMID- 19307421 TI - Mechanisms mediating brain plasticity: IGF1 and adult hippocampal neurogenesis. AB - This review addresses the role of serum insulin-like growth factor 1 (IGF1) as one mechanism of adult neural plasticity, specifically, its regulation of hippocampal neurogenesis among other plasticity-related processes. It is suggested that IGF has been reused advantageously both for the control of energy expenditure as a function of the organism's activity and to protect, repair, and plastically modulate the brain. Moreover, because as the main source of IGF1 in the adult organism is outside the brain and its presence in this organ is a function of the activity, IGF1 becomes an ideal factor to induce plastic/neuroprotective functions as a function of the organism's activity. The link for this point of view comes from the original function of IGF1 during ontogeny/phylogeny, the promotion of cell survival and control of neural cell numbers, whereas one of the IGF1 functions in the adult brain is the control of hippocampal neurogenesis. The investigation of the IGF1 role as mediator of exercise effects suggests that many but not all the effects of physical activity are mediated by IGF1. These investigations have contributed to delimit the role of IGF1 as mediator of exercise actions, but at the same time are unveiling new roles for serum IGF1 inside the brain. PMID- 19307422 TI - Synaptic plasticity, neurogenesis, and functional recovery after spinal cord injury. AB - Spinal cord injury research has greatly expanded in recent years, but our understanding of the mechanisms that underlie the functional recovery that can occur over the weeks and months following the initial injury, is far from complete. To grasp the scope of the problem, it is important to begin by defining the sensorimotor pathways that might be involved by a spinal injury. This is done in the rodent and nonhuman primate, which are two of the most commonly used animal models in basic and translational spinal injury research. Many of the better known experimentally induced models are then reviewed in terms of the pathways they involve and the reorganization and recovery that have been shown to follow. The better understood neuronal mechanisms mediating such post-injury plasticity, including dendritic spine growth and axonal sprouting, are then examined. PMID- 19307423 TI - Individual differences in episodic memory: the role of self-initiated encoding strategies. AB - Individuals' abilities to form and retrieve episodic memories vary widely. Consistent with this, there are substantial individual differences in brain activity during encoding and retrieval that are associated with individual differences in memory performance. Growing evidence suggests that individual differences in self-initiated encoding strategy use play an important role in individual differences in episodic memory and brain activity during intentional encoding. This review examines the role of individual differences in self initiated encoding strategy use in individual differences in episodic memory, and outlines the major findings of brain lesion and functional neuroimaging studies that characterize the neural correlates of individual differences in self initiated encoding strategy use. The relevance of individual differences in self initiated encoding strategy use to understanding episodic memory impairments and alterations in brain activity in clinical populations such as individuals with schizophrenia is also discussed. PMID- 19307424 TI - Agrin, aquaporin-4, and astrocyte polarity as an important feature of the blood brain barrier. AB - The blood-brain barrier (BBB) does not exclusively refer to brain endothelial cells, which are the site of the barrier proper. In the past few years, it has become increasingly clear that BBB endothelial cells depend considerably on the brain microenvironment to a degree exceeding the environmental influence in other organs. The concept of the BBB has been continuously developed over the decades, culminating now in the recognition that endothelial cell function in the brain is not limited to simply mediating energy and oxygen transfer between blood and neural tissue. Endothelial cells are rather "Janus-headed beings" that are active partners of both luminal molecules and cells, as well as subendothelial cells such as pericytes, astrocytes, and neurons. In this overview, the authors present and discuss both the role of astroglial cells in managing the BBB and aspects of pathological alterations in the brain as far as the BBB is involved. After a brief introduction of the BBB that describes the structure and function of the brain capillary endothelial cells, the authors report on both the water channel protein aquaporin-4 (AQP4) in astrocytes and the extracellular matrix between astrocytes/pericytes and endothelial cells. The AQP4 has an important impact on the homeostasis in the brain parenchyma; however, the mechanistic cascade from the composition of the astrocyte membrane to the maintenance of BBB properties in the endothelial cells, including their tight junction formation, is still completely unknown. PMID- 19307425 TI - Respiratory circuits: function, mechanisms, topology, and pathology. AB - Neuroscientists have long sought to understand how circuits in the nervous system are organized to generate the precise neural outputs that underlie particular behaviors. Recent studies deepened our understanding of the mechanisms responsible for the generation of the rhythmic output for breathing. Here, the author focuses on issues that are pertinent for the respiratory network and considers its organization and how it derives the functional output. The author discusses pacemaker and network mechanisms of rhythm generation, which are now combined into a novel concept of emergent network activity due to coherent excitation of pacemaker groups. He discusses subcellular basis of this hypothesis and possible mechanisms of synchronization within respiratory network. These new findings in respiratory neuroscience are further applied to explain modifications in breathing during hypoxia and possible origins of respiratory disorders that may be acquired during neural development and aging. PMID- 19307426 TI - Cell cycle regulatory control for uterine stromal cell decidualization in implantation. AB - Uterine stromal cell decidualization is integral to successful embryo implantation, which is a gateway to pregnancy establishment. This process is characterized by stromal cell proliferation and differentiation into decidual cells with polyploidy. The molecular mechanisms that are involved in these events remain poorly understood. The current concept is that locally induced factors with the onset of implantation influence uterine stromal cell proliferation and/or differentiation through modulation of core cell cycle regulators. This review will aim to address the currently available knowledge on interaction between growth factor/homeobox and cell cycle regulatory signaling in the progression of various aspects of decidualization. PMID- 19307427 TI - Transforming growth factor-beta1 inhibits luteinization and promotes apoptosis in bovine granulosa cells. AB - We have previously shown that TGFB1 inhibits estradiol (E(2)) and progesterone (P(4)) biosynthesis in FSH-stimulated bovine granulosa cells by selective inhibition of steroidogenic enzymes. The objective of this study was to assess the effects of TGFB1 on E(2) and P(4) steroidogenesis in bovine granulosa cells cultured in the absence of FSH and to measure the effects of TGFB1 on cell proliferation and apoptosis in the presence and absence of FSH. Bovine granulosa cells from 2 to 5 mm follicles were cultured in serum-free medium for 2-6 days. In the absence of FSH, the secretion of P(4) increased with time in culture (P<0.05). Addition of TGFB1 for 6 days decreased P(4) secretion and mRNA levels of the P(4) synthesis-associated genes STAR, CYP11A1, HSD3B1, and GSTA (P<0.05). In the absence of FSH, the secretion of E(2) decreased and addition of TGFB1 for 6 days partially reversed this decline and stimulated E(2) biosynthesis, CYP19A1 and HSD17B1 mRNA levels and CYP19A1 activity (P<0.05). Conversely, TGFB1 did not affect HSD17B7 expression and HSD17B-reducing activity. TGFB1 decreased the proportion of cells in the G0/G1 and S+G2/M phases in FSH-stimulated and unstimulated granulosa cells (P<0.05). Furthermore, in the presence or absence of FSH, TGFB1 increased the proportion of cells in apoptosis measured by propidium iodide staining and flow cytometry and confirmed by increased levels of cleaved caspase-3 (P<0.05). Our results therefore indicate that TGFB1 inhibits luteinization in cultured bovine granulosa cells while maintaining an estrogenic phenotype, and this effect was associated with increased apoptosis. PMID- 19307432 TI - Taking chances in the face of threat: romantic risk regulation and approach motivation. AB - Four studies examine the hypothesis that goals adopted by high and low self esteem people (HSEs and LSEs) to manage risk in romantic relationships may reflect global shifts in approach motivation and subsequently affect risk taking in nonsocial domains. In Studies 1 and 2, threats to participants' romantic relationships heightened HSEs' self-reported general approach motivation while lowering LSEs' approach motivation. In Studies 2 through 4, HSEs exhibited riskier decision making (i.e., a greater tendency to pursue rewards and ignore risks) in nonsocial domains following a relationship threat manipulation whereas LSEs made more conservative decisions. These results suggest that the romantic risk regulation may be inherently linked to a broader approach and avoidance system and that specific risk regulation behaviors may be driven by global motivational shifts to a greater degree than previously theorized. PMID- 19307433 TI - Evaluating one performance among others: the influence of rank and degree of exposure to comparison referents. AB - Three studies examined the influence of comparison-referent exposure (i.e., the frequency with which one views comparison referents) on evaluations of the ability of a target person (either oneself or another person). In Experiment 1, participants performed a task and then viewed performances of both upward and downward referents. Participants who saw more performances by the upward referents than the downward referents evaluated their own performances less favorably did than participants who saw more performances by the downward referents than the upward referents. Experiment 2 produced similar findings, showing that comparison exposure also influences people's evaluations about someone other than themselves. In Experiment 3, comparison-exposure effects were significantly reduced when participants were instructed to think deliberatively about the comparison information, consistent with the idea that people typically rely on imprecise representations of comparison information even when they are capable of forming more precise representations from memory if motivated to do so. PMID- 19307434 TI - Split-belt treadmill adaptation transfers to overground walking in persons poststroke. AB - BACKGROUND AND OBJECTIVE: Following stroke, subjects retain the ability to adapt interlimb symmetry on the split-belt treadmill. Critical to advancing our understanding of locomotor adaptation and its usefulness in rehabilitation is discerning whether adaptive effects observed on a treadmill transfer to walking over ground. We examined whether aftereffects following split-belt treadmill adaptation transfer to overground walking in healthy persons and those poststroke. METHODS: Eleven poststroke and 11 age-matched and gender-matched healthy subjects walked over ground before and after walking on a split-belt treadmill. Adaptation and aftereffects in step length and double support time were calculated. RESULTS: Both groups demonstrated partial transfer of the aftereffects observed on the treadmill (P<.001) to overground walking (P<.05), but the transfer was more robust in the subjects poststroke (P<.05). The subjects with baseline asymmetry after stroke improved in asymmetry of step length and double limb support (P=.06). CONCLUSIONS: The partial transfer of aftereffects to overground walking suggests that some shared neural circuits that control locomotion for different environmental contexts are adapted during split-belt treadmill walking. The larger adaptation transfer from the treadmill to overground walking in the stroke survivors may be due to difficulty adjusting their walking pattern to changing environmental demands. Such difficulties with context switching have been considered detrimental to function poststroke. However, we propose that the persistence of improved symmetry when changing context to overground walking could be used to advantage in poststroke rehabilitation. PMID- 19307435 TI - Rhythm perturbations in acoustically paced treadmill walking after stroke. AB - BACKGROUND: In rehabilitation, acoustic rhythms are often used to improve gait after stroke. Acoustic cueing may enhance gait coordination by creating a stable coupling between heel strikes and metronome beats and provide a means to train the adaptability of gait coordination to environmental changes, as required in everyday life ambulation. OBJECTIVE: To examine the stability and adaptability of auditory-motor synchronization in acoustically paced treadmill walking in stroke patients. METHODS: Eleven stroke patients and 10 healthy controls walked on a treadmill at preferred speed and cadence under no metronome, single-metronome (pacing only paretic or nonparetic steps), and double-metronome (pacing both footfalls) conditions. The stability of auditory-motor synchronization was quantified by the variability of the phase relation between footfalls and beats. In a separate session, the acoustic rhythms were perturbed and adaptations to restore auditory-motor synchronization were quantified. RESULTS: For both groups, auditory-motor synchronization was more stable for double-metronome than single metronome conditions, with stroke patients exhibiting an overall weaker coupling of footfalls to metronome beats than controls. The recovery characteristics following rhythm perturbations corroborated the stability findings and further revealed that stroke patients had difficulty in accelerating their steps and instead preferred a slower-step response to restore synchronization. CONCLUSIONS: In gait rehabilitation practice, the use of acoustic rhythms may be more effective when both footfalls are paced. In addition, rhythm perturbations during acoustically paced treadmill walking may not only be employed to evaluate the stability of auditory-motor synchronization but also have promising implications for evaluation and training of gait adaptations in neurorehabilitation practice. PMID- 19307436 TI - The VEnous INtervention (VEIN) Project. PMID- 19307437 TI - Interventions for uncomplicated varicose veins. AB - OBJECTIVE: To review the evidence regarding interventions (compression, sclerotherapy, surgery and endoluminal) for uncomplicated venous disease. METHOD: A literature search of MEDLINE and EMBASE was performed. In addition, bibliographies of published data and the Cochrane Peripheral Vascular Review Group Specialist Register were examined. Publications describing an intervention for uncomplicated venous disease or trials comparing one intervention against another were considered. RESULTS: Uncomplicated but symptomatic varicose veins are associated with a significant reduction in quality of life (QoL). Evaluation must include a detailed history and examination supported by non-invasive imaging. Although HHD is useful, its utility is limited in the popliteal fossa. It can not provide morphological/anatomical detail when considering an endoluminal treatment option. Regardless of intervention, all treatments considered are associated with a significant improvement in QoL at acceptable cost. CONCLUSION: Uncomplicated symptomatic varicose veins lead to a reduced QoL, which can be significantly improved by all interventions considered. PMID- 19307438 TI - A systematic review of compression hosiery for uncomplicated varicose veins. AB - OBJECTIVE: Compression hosiery is widely used in the prevention and management of symptoms related to varicose veins. However, there are still gaps and questions in relation to its benefit. This review seeks to examine the current evidence regarding the effectiveness of compression hosiery in the treatment of varicose veins. METHOD: Prospective, randomized controlled trials (RCTs) evaluating compression hosiery in the treatment of varicose veins were sought. Where RCTs were unavailable other evidence was included. Studies were included if they evaluated the application of compression to patients with a diagnosis of varicose veins. Twelve electronic bibliographic databases and 18 internet-based research resources were searched. Inclusion or exclusion of trials was decided by two reviewers acting independently. RESULTS: The search strategy identified 25 studies. Eleven were RCTs or systematic reviews, 12 non-randomized studies and two guidelines. No consensus was found regarding the class of compression needed for the effective management of varicose veins. Wearing compression improved symptom management, but could be confounded by the exclusion of high number of non-compliant patients within the trials. Wearing compression to slow the progression, or prevent the reoccurrence of varicose veins could not be supported by the current published evidence. CONCLUSION: The evidence for the benefit of compression hosiery for varicose veins was equivocal. The published literature was often contradictory and had methodological flaws. PMID- 19307439 TI - Standard varicose vein surgery. AB - This article examines the practice of standard varicose vein surgery including sapheno-femoral and sapheno-popliteal ligation, perforator surgery and surgery for recurrent varicose veins. The technique of exposure of the sapheno-femoral junction and the sapheno-popliteal junction is outlined and advice given on avoidance of complications for both. The evidence regarding methods of closure over the ligated sapheno-femoral junction is examined as is the requirement for stripping and the use of different types of stripper. The requirement to strip the small saphenous vein and the extent of dissection necessary in the popliteal fossa is also examined. Complications of standard varicose vein surgery are outlined. The frequency of wound infection, nerve injury, vascular injury and venous thromboembolism are listed and strategies to avoid these complications are examined. PMID- 19307440 TI - Radiofrequency ablation for uncomplicated varicose veins. AB - In recent years, minimally invasive endovenous treatments have gained popularity in the treatment of superficial venous reflux. The perceived advantages of endovenous therapy include reduced pain, high vein occlusion rates, and early return to work and normal activities. Endovenous radiofrequency ablation (RFA) involves the delivery of thermal energy from a bipolar catheter to the venous segment to be treated. This technique has been available since 1998 and numerous devices and catheters are now produced. Numerous prospective and randomized studies have compared the effectiveness of RFA with traditional and endovenous procedures. In this article, the available evidence for clinical effectiveness, quality of life and cost gains following endovenous RFA is summarized. The scientific principles behind RFA and technical procedural considerations are discussed and standards of care for the delivery of endovenous RFA are proposed. PMID- 19307441 TI - Endovenous laser treatment for uncomplicated varicose veins. AB - OBJECTIVE: Endovenous laser ablation (EVLA) of incompetent truncal veins has been proposed as a minimally invasive alternative to conventional surgery for varicose veins. Various strategies have been proposed for successful treatment and this study reviews the evidence for these. METHOD: A Medline and 'controlled trials online database' search was performed to identify original articles and randomized controlled trials (RCTs) reporting outcomes for EVLA. Information on patient selection, equipment, technique and outcomes were recorded. RESULTS: Ninety-eight original studies, including five RCTs, were identified. RCT data indicate short-term outcomes (abolition of reflux, improvement in quality of life [QOL], patient satisfaction) were equivalent to those for surgery. Long-term follow-up is not available. A further RCT showed superior outcomes for ablation commencing at the lowest point of superficial venous reflux rather than at an arbitrary point (fewer residual varicosities, greater improvement in QOL). Non randomized series suggest that laser energy of >60 J/cm results in reliable truncal vein occlusion and that longer wavelength lasers may be associated with less post-treatment discomfort. CONCLUSION: In the short-term EVLA is a safe and effective treatment for patients with varicose veins. Long-term follow-up is still required. PMID- 19307442 TI - Foam and liquid sclerotherapy for varicose veins. AB - OBJECTIVE: The objective of this study is to review the methods and outcome of ultrasound-guided foam sclerotherapy (UGFS) for the treatment of superficial venous incompetence. METHOD: Medical literature databases including Medline were searched for recent literature concerning UGFS. Papers describing methods and outcome have been assessed and their main findings included in this summary. A detailed description of the methods used by the author has been included as an example of how successful the treatment may be achieved. RESULTS: A diverse range of practice is described in published literature in this field. Each group of authors used their own variation of the methods, described in the published literature, with good results. It is clear that foam sclerotherapy is far more effective than liquid sclerotherapy and that ultrasound imaging allows the treatment to be delivered accurately to affected veins. There is evidence that 3% policocanol foam is no more effective than 1% polidocanol foam. The optimum ratio of gas to liquid is 4:1, although a range of ratios is reported in published work. There is a wide variation in the volume used as well as the method by which it is injected. The use of carbon dioxide foam reduces the systemic complications, particularly visual disturbance, when compared with air foams. Very few serious adverse events have been reported in the literature despite the widespread use of this method. Rates of recanalization of saphenous trunks following UGFS are similar to those observed after endovenous laser and endovenous radiofrequency ablation of veins, as well as the residual incompetence after surgical treatment. CONCLUSIONS: UGFS is a safe and effective method of treating varicose veins. The relative advantages or disadvantages of this treatment in the longer term are yet to be published. PMID- 19307443 TI - Association between smoking and retrospectively reported attention deficit/hyperactivity disorder symptoms in a large sample of new mothers. AB - INTRODUCTION: This study investigated the association between retrospectively reported attention-deficit/hyperactivity disorder (ADHD) symptoms experienced during childhood and five cigarette smoking-related outcomes in adulthood. METHODS: A large sample (N = 1,117) of new mothers participating in an ongoing longitudinal study completed retrospective reports of their childhood ADHD symptomatology, as well as concurrent and retrospective reports of their smoking behavior. Linear regression models tested the association between ADHD symptomatology and smoking outcomes. RESULTS: Childhood ADHD symptomatology was predictive of the number of cigarettes smoked per day currently and during pregnancy, as well as the age at onset of smoking. We found nonlinear associations between hyperactive-impulsive symptoms and the number of cigarettes smoked per day in pregnancy, as well as between inattentive symptoms and the number of cigarettes smoked per day currently. Women who retrospectively reported intermediate levels of ADHD symptoms during their childhood reported smoking more cigarettes per day than women who reported low or high levels of ADHD symptoms during childhood. We also found multiplicative relationship between inattentive and hyperactive-impulsive symptoms, such that inattentive symptoms were predictive of an earlier age at smoking onset only when hyperactive-impulsive symptoms were low; moreover, the magnitude of this association was stronger for Black relative to White women. DISCUSSION: These findings demonstrate the importance of considering differential effects of ADHD symptoms and smoking outcomes as a function of sex and race. They also represent a potentially indirect means through which women who have even a moderate childhood history of ADHD symptomatology may create a set of circumstances that compromise the health and well-being of their own children. PMID- 19307444 TI - Examination of the nicotine dependence (NICSNP) consortium findings in the Iowa adoption studies population. AB - INTRODUCTION: Nicotine dependence results from a complex interplay of genetic and environmental factors. Over the past several years, a large number of studies have been performed to identify distinct gene loci containing genetic vulnerability to nicotine dependence. Two of the most prominent studies were conducted by the Collaborative Study of the Genetics of Nicotine Dependence (NICSNP) Consortium using both candidate gene and high-density association approaches. METHODS: We attempted to confirm and extend the most significant findings from the high-density association study and the candidate gene study using the behavioral and genetic resources of the Iowa Adoption Studies, the largest case-control adoption study of substance use in the United States. RESULTS: We found evidence that genetic variation at CHRNA1, CHRNA2, CHRNA7, and CHRNB1 alters susceptibility to nicotine dependence, but we did not replicate any of the most significant single nucleotide polymorphism associations from the NICSNP high-density association study. DISCUSSION: Further examination of the NICSNP findings in other population samples is indicated. PMID- 19307445 TI - Use of propensity score matching in evaluating smokeless tobacco as a gateway to smoking. AB - INTRODUCTION: The contentious debate over the promotion of Swedish snus, a form of moist snuff, as an alternative to cigarettes has often focused on the product's potential as a gateway to smoking. Findings from prospective studies in the United States have suggested that smokeless tobacco (SLT) is a significant predictor of smoking onset, before and after adjustment for baseline covariates. Use of conventional regression methods in these studies may have resulted in biased parameter estimates, arising from imbalanced covariate distributions in the users and nonusers of SLT. An alternative approach, which has been used widely in the econometric literature, matches exposure or treatment levels on the basis of the propensity score distribution. METHODS: Using this approach, we matched current SLT users from the National Longitudinal Study of Adolescent Health with nonusers (496 pairs) and followed them from adolescence into young adulthood for determination of smoking status. RESULTS: Prior to matching, the unadjusted risk of becoming a daily smoker was significantly greater for the SLT users compared with nonusers (n = 10,820; range of relative risk = 1.3-2.0, p < .001). However, after pairing individuals on propensity score, we found no evidence for an increased risk of smoking among the SLT users. DISCUSSION: Baseline differences in the risk factors for smoking likely account for the association between the two tobacco products. PMID- 19307446 TI - Examining unsupervised time with peers and the role of association with delinquent peers on adolescent smoking. AB - INTRODUCTION: This study explored the association between unsupervised time with peers and adolescent smoking behavior both directly and indirectly through interaction with delinquent peers, social expectancies about cigarette smoking, and cigarette offers from peers. METHODS: A cross-sectional survey was used for the study and included 248 male and female middle school students. RESULTS: Results of structural equation modeling revealed that unsupervised time with peers is associated indirectly with adolescent smoking behavior through the mediation of association with delinquent peers, social expectancies about cigarette smoking, and cigarette offers from peers. DISCUSSION: Interventions designed to motivate adolescents without adult supervision to associate more with friends who engage in prosocial activities may eventually reduce adolescent smoking. Further implications for structured supervised time for students outside of school time are discussed. PMID- 19307447 TI - Targeting pregnant and parental smokers: long-term outcomes of a practice-based intervention. AB - INTRODUCTION: This study aimed to assess the change in obstetric and pediatric provider smoking cessation practices following implementation of a practice guideline-driven office-based program. METHODS: This pre-post evaluation took place between May 2003 and August 2006 in 1 pediatric and 1 obstetric hospital based clinic. The intervention involved provider training combined with office system supports. A total of 1,080 exit interviews were collected to measure outcomes of clinic practices at baseline and at 1 month, 6 months, 1 year (obstetric), and 2 years (pediatric) after implementation. Trend analysis was used to assess change in practice rates over time. RESULTS: Following program implementation, pediatric provider "Ask" rates increased (49% before to 86% 2 years after, p < .0001); changes in pediatric "Advise" and "Assist" rates were not significant: 44%-59% (p = .19) and 18%-28% (p = .26), respectively. In the obstetric clinic, whereas no significant changes were detected in provider "Ask" (59%-65% 1 year after, p = .17) or "Advise" (72%-85%, p = .27) rates, "Assist" rates rose from 28% to 62% (p = .0075) 1 year after program implementation. DISCUSSION: Implementation of the office-based program achieved significantly improved trends in pediatric provider "Ask" rates and obstetric provider "Assist" rates over time. Further research is needed on office strategies to create long term provider behavior changes in smoking cessation practices. PMID- 19307448 TI - Abstracts of the British Society of Gastroenterology Annual General Meeting. March 23-26, 2009. Glasgow, Scotland. PMID- 19307449 TI - Differential localization of flavin-containing monooxygenase (FMO) isoforms 1, 3, and 4 in rat liver and kidney and evidence for expression of FMO4 in mouse, rat, and human liver and kidney microsomes. AB - Flavin-containing monooxygenases (FMOs) play significant roles in the metabolism of drugs and endogenous or foreign compounds. In this study, the regional distribution of FMO isoforms 1, 3, and 4 was investigated in male Sprague-Dawley rat liver and kidney using immunohistochemistry (IHC). Rabbit polyclonal antibodies to rat FMO1 and FMO4, developed using anti-peptide technology, and commercial anti-human FMO3 antibody were used; specificities of the antibodies were verified using Western blotting, immunoprecipitation, and IHC. In liver, the highest immunoreactivity for FMO1 and FMO3 was detected in the perivenous region, and immunoreactivity decreased in intensity toward the periportal region. In contrast, FMO4 immunoreactivity was detected with the opposite lobular distribution. In the kidney, the highest immunoreactivity for FMO1, -3, and -4 was detected in the distal tubules. FMO1 and FMO4 immunoreactivity was also detected in the proximal tubules with strong staining in the brush borders, whereas less FMO3 immunoreactivity was detected in the proximal tubules. Immunoreactivity for FMO3 and FMO4 was detected in the collecting tubules in the renal medulla and the glomerulus, whereas little FMO1 immunoreactivity was detected in these regions. The FMO1 antibody did not react with human liver or kidney microsomes. However, the FMO4 antibody reacted with male and female mouse and human tissues. These data provided a compelling visual demonstration of the isoform-specific localization patterns of FMO1, -3, and -4 in the rat liver and kidney and the first evidence for expression of FMO4 at the protein level in mouse and human liver and kidney microsomes. PMID- 19307450 TI - Characterization of mouse heart adenylyl cyclase. AB - Chronic heart failure is one of the most frequent causes of death in humans. Knockout of type 5 adenylyl cyclase (AC) in mice causes longevity and protection from cardiomyopathy, and an AC5 inhibitor reduces beta-adrenoceptor-stimulated Ca(2+) inward currents in isolated mouse cardiomyocytes. These data indicate that selective AC5 inhibitors may be beneficial in chronic heart failure. Therefore, we characterized AC in mouse heart membranes. Real-time polymerase chain reaction and immunoblot analysis suggested that AC5 is an important heart AC isoform. Enzyme kinetics of heart AC and recombinant AC5 in the presence of Mg(2+) were similar. Moreover, the inhibitory profile of eight 2'(3')-O-(N methylanthraniloyl) (MANT)-nucleoside 5'-([gamma-thio])triphosphates on mouse heart in the presence of Mg(2+) was almost identical to that of AC5. MANT-ITP was the most potent inhibitor of heart AC and recombinant AC5, with K(i) values in the 15 to 25 nM range in the presence of Mg(2+) and in the 1 to 5 nM range in the presence of Mn(2+). However, in the presence of Mn(2+), we also noted differences between mouse heart AC and AC5 with respect to enzyme kinetics and forskolin analog effects. In conclusion, with regard to expression and kinetics and inhibition by MANT-nucleotides in the presence of Mg(2+), AC5 is an important AC isoform in heart, with MANT-ITP being an excellent starting point for the design of AC5-selective inhibitors. Unfortunately, a limitation of our study is the fact that immunologically and biochemically, AC5 and AC6 are quite similar, although they have different roles in heart. Moreover, lack of antibody specificity and Mn(2+) masking AC5 effects were problems. PMID- 19307451 TI - Erythropoietin protects the heart from ventricular arrhythmia during ischemia and reperfusion via neuronal nitric-oxide synthase. AB - Erythropoietin (EPO) is a potent cardioprotective agent in models of myocardial ischemia and reperfusion (I/R). It has been suggested recently that EPO may also reduce ventricular arrhythmia after I/R. The present study investigated the role of neuronal nitric oxide synthase (nNOS) on the antiarrhythmic effects of EPO. EPO treatment increased nNOS expression in isolated neonatal mouse ventricular myocytes. Cotreatment with the phosphatidylinositol 3 (PI3)-kinase inhibitor, LY294002 [2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride], or treatment of cardiomyocytes infected with a dominant negative adenovirus targeted to Akt1 (ADV-dnAkt1) blocked the effects of EPO on nNOS expression, suggesting that EPO regulates nNOS expression via PI3-kinase and Akt. To examine the in vivo antiarrhythmic effects of EPO, wild-type (WT) and nNOS(-/-) mice were anesthetized and, after a baseline measurement, subjected to myocardial I/R to provoke ventricular arrhythmias. Pretreatment with EPO 24 h before ischemia increased nNOS expression and significantly reduced the number of premature ventricular contractions (PVCs) and the incidence of ventricular tachycardia (VT) in WT mice. In contrast, treatment with EPO had no effect on PVCs or the incidence of VT in nNOS(-/-) mice. Furthermore, EPO treatment after ischemia significantly reduced the threshold dose of cesium chloride (CsCl) to induce VT. We conclude that EPO via nNOS protects the heart from spontaneous and CsCl induced ventricular arrhythmia during myocardial I/R. PMID- 19307452 TI - Sepiapterin decreases acute rejection and apoptosis in cardiac transplants independently of changes in nitric oxide and inducible nitric-oxide synthase dimerization. AB - Tetrahydrobiopterin (BH(4)), a cofactor of inducible nitric-oxide synthase (iNOS), is an important post-translational regulator of NO bioactivity. We examined whether treatment of cardiac allograft recipients with sepiapterin [S-( )-2-amino-7,8-dihydro-6-(2-hydroxy-1-oxopropyl)-4-(1H)-pteridinone], a precursor of BH(4), inhibited acute rejection and apoptosis in cardiac transplants. Heterotopic cardiac transplantation was performed in Wistar-Furth donor to Lewis recipient strain rats. Recipients were treated daily after transplantation with 10 mg/kg sepiapterin. Grafts were harvested on post-transplant day 6 for analysis of BH(4) (high-performance liquid chromatography), expression of inflammatory cytokines (reverse transcription- and real-time polymerase chain reaction), iNOS (Western blots), and NO (Griess reaction and NO analyzer). Histological rejection grade was scored, and graft function was determined by echocardiography. Apoptosis, protein nitration, and oxidative stress were determined by immunohistochemistry. Treatment of allografts with sepiapterin increased cardiac BH(4) levels by 3-fold without changing protein levels of GTP cyclohydrolase, the enzyme that regulates de novo BH(4) synthesis. Sepiapterin decreased inflammatory cell infiltrate and significantly inhibited histological rejection scores and apoptosis similar in magnitude to cyclosporine. Sepiapterin also decreased nitrative and oxidative stress. Sepiapterin caused a smaller increase in left ventricular mass versus untreated allografts but without improving fractional shortening. Sepiapterin did not alter tumor necrosis factor-alpha and interferon gamma expression, whereas it decreased interleukin (IL)-2 expression. Sepiapterin did not change total iNOS protein or monomer levels, or plasma and tissue NO metabolites levels. It is concluded that the mechanism(s) of antirejection are due in part to decreased apoptosis, protein nitration, and oxidation of cardiomyocytes, which seems to be mediated at the immune level by limiting inflammatory cell infiltration via decreased IL-2-mediated T-lymphocyte expansion. PMID- 19307453 TI - A force for social change. PMID- 19307454 TI - Commentary on "the brief serenity scale: a psychometric analysis of a measure of spirituality and well-being". PMID- 19307455 TI - Commentary on "the provision of spiritual care by registered nurses on a maternal -infant unit". PMID- 19307457 TI - Commentary on "existential and spiritual needs in mental health care: an ethical and holistic perspective". PMID- 19307458 TI - In vitro and in vivo modulation of cartilage degradation by a standardized Centella asiatica fraction. AB - Osteoarthritis (OA) is a degenerative joint disease in which focal cartilage destruction is one of the primary features. The present study aims to evaluate the effect of a Centella asiatica fraction on in vitro and in vivo cartilage degradation. Bovine cartilage explants and bovine chondrocytes cultured in alginate were stimulated with IL-1 beta in the presence or absence of different concentrations (2, 5 and 10 microg/ml) of a standardized Centella asiatica triterpenes (CAT) fraction. The CAT fraction inhibited the IL-1 beta-induced proteoglycan (PG) release and nitric oxide (NO) production by cartilage explants in a dose-dependent manner. The IL-1 beta-induced reduction in PG synthesis and proliferation of chondrocytes cultured in alginate were counteracted by the CAT fraction at a concentration of 10 microg/ml. In a zymosan-induced acute arthritis model, the CAT fraction inhibited PG depletion without modulating joint swelling and inflammatory cell infiltration. In conclusion, the present study demonstrated for the first time that the tested Centella asiatica fraction was able to inhibit the zymosan-induced cartilage degradation in vivo without affecting the zymosan induced inflammatory cell infiltration and joint swelling. The in vitro data indicate that the cartilage protective activity might at least partially be induced by the inhibition of NO production. The overall results indicate a possible disease modifying osteoarthritic activity of the Centella asiatica fraction. PMID- 19307459 TI - Increased CYP4B1 mRNA is associated with the inhibition of dextran sulfate sodium induced colitis by caffeic acid in mice. AB - Susceptibility to inflammatory bowel diseases depends upon interactions between the genetics of the individual and induction of chronic mucosal inflammation. We hypothesized that administration of dietary phenolics, caffeic acid and rutin, would suppress upregulation of inflammatory markers and intestinal damage in a mouse model of colitis. Colitis was induced in C3H/ HeOuJ mice (8 weeks old, 6 male/6 female per treatment) with 1.25% dextran sulfate sodium (DSS) for 6 d in their drinking water. Rutin (1.0 mmol (524 mg)/kg in diet), caffeic acid (1.0 mmol (179 mg)/kg in diet), and hypoxoside extract (15 mg/d, an anticolitic phenolic control) were fed to the mice for 7 d before and during DSS treatment, as well as without DSS treatment. Body weight loss was prevented by rutin and caffeic acid during DSS treatment. Colon lengths in mice fed caffeic acid and hypoxoside during DSS treatment were similar to DSS-negative control. Food intake was improved and myeloperoxidase (MPO) was decreased with each phenolic treatment in DSS-treated mice compared with DSS treatment alone. Colonic mRNA expression of IL-17 and iNOS were inhibited when IL-4 was increased by each phenolic treatment combined with DSS, whereas CYP4B1 mRNA was increased only by caffeic acid in DSS treated mice, compared with DSS treatment alone. Colonic and cecal histopathology scores of DSS-treated mice were significantly more severe (P < 0.01) than in mice fed caffeic acid before and during DSS treatment, based on mucosal height, necrosis, edema, erosion, and inflammatory cell infiltration. Although both rutin and caffeic acid suppressed the expression of selected inflammatory markers, only caffeic acid protected against DSS-induced colitis, in association with normalization of CYP4B1 expression. The inhibition of DSS-induced colitic pathology by caffeic acid was mediated by mechanisms in addition to anti inflammatory effects that deserve further study. PMID- 19307460 TI - Vertebrae of developing fat-1 mice have greater strength and lower n-6/n-3 fatty acid ratio. AB - Incorporation of dietary n-3 polyunsaturated fatty acids (PUFA) into bone may optimize bone development. The study objective was to use the fat-1 mouse, a transgenic model that synthesizes n-3 PUFA from n-6 PUFA, to determine if bone mineral density (BMD) and biomechanical bone strength were favourably modulated by lowering the n-6/n-3 PUFA ratio in vertebrae. Male and female wild-type and fat-1 mice were fed an AIN93-G diet containing 10% safflower oil from weaning through 12 weeks of age. Vertebrae BMD was determined by dual energy x-ray absorptiometry and peak load, a surrogate measure of fracture risk, was measured by a materials testing system. Vertebrae fatty acid composition was measured by gas liquid chromatography. At 12 weeks of age, vertebrae peak load was higher in fat-1 mice compared to wild-type (P = 0.026). Fat-1 mice also had lower n-6/n-3 PUFA ratio in vertebrae than wild-type (P < 0.001) and this ratio was negatively correlated with BMD and peak load (P = 0.005). Moreover, n-3 PUFA including alpha linolenic acid, eicosapentaenoic acid and docosahexaenoic acid were positively correlated (P < 0.05) with BMD and peak load. Therefore, a lower vertebrae n-6/n 3 PUFA ratio is associated with stronger vertebrae and suggests a positive role for n-3 PUFA in bone development. PMID- 19307461 TI - Improvement in dissolution of liver fibrosis in an animal model by tetrathiomolybdate. AB - The background for this study is that we have observed some improvement in cirrhosis in Wilson's disease patients treated with the anticopper medicine, zinc, and another anticopper drug, tetrathiomolybdate, has completely prevented hepatic fibrosis in the carbon tetrachloride mouse model. We hypothesize that in existing cirrhosis, there may be a fine balance between fibrosis formation and fibrosis dissolution, which may be pushed in the direction of dissolution by anticopper drugs. Thus, in this study, we produced hepatic fibrosis in mice by treatment with carbon tetrachloride, then gave half the fibrotic mice tetrathiomolybdate for 3 months, while the other half of the fibrotic mice received nothing for 3 months and served as controls. Tetrathiomolybdate caused a dramatic and significant reduction in fibrosis as measured by hydroxyproline (the major amino acid constituent of collagen) levels, almost back to baseline levels, compared to controls, who had only a slight and nonsignificant reduction. It is clear from this animal study that dissolution of preexisting fibrosis can be strongly catalyzed by lowering copper levels with tetrathiomolybdate. It now becomes important to evaluate whether this approach will work in the human epidemic of cirrhotic disease resulting from diseases such as alcoholism, nonalcoholic steatohepatitis, and hepatitis C. PMID- 19307462 TI - Interaction between TRPC1/TRPC4 assembly and STIM1 contributes to store-operated Ca2+ entry in mesangial cells. AB - Although Orai1 protein was recently identified as the component of CRAC channels in hematopoietic cells, store-operated channels (SOC) in other cell types may have a different molecular entity. Also, the activation mechanism of SOC remains unclear, in general. In the present study, we tested the hypothesis that TRPC1 and TRPC4 proteins were functional subunits of SOC in glomerular mesangial cells (MCs) and that STIM1 was required for the channel activation through interaction with the TRPC proteins. In cultured human MCs, cell-attached patch clamp and fura 2 fluorescence measurements showed that single knockdown of either TRPC1 or TRPC4 significantly attenuated thapsigargin-induced membrane currents and Ca2+ entry as well as Ang II-induced channel activity. Double knockdown of both TRPCs resulted in a comparable inhibition on store-operated Ca2+ entry with single knockdown of either TRPC. Consistent with our previous report, co-immunoprecipitation showed a physical interaction between TRPC1 and TRPC4. Furthermore, we found that knockdown of STIM1 using RNAi significantly reduced the thapsigargin-stimulated membrane currents. Co-immunoprecipitation showed that STIM1 interacted with TRPC4, but not TRPC1. In addition, simultaneous inhibition of STIM1 and TRPC1 resulted in a comparable effect on SOC with single inhibition of either one of them. Taken together, we conclude that in glomerular mesangial cells, the TRPC1/TRPC4 complexes constitute the functional subunits of SOC and that the interaction between STIM1 and TRPC4 may be the mechanism for the activation of the channels. PMID- 19307463 TI - Modulation of iron-regulatory genes in human hepatocellular carcinoma and its physiological consequences. AB - Hepatocellular carcinoma (HCC) commonly develops in patients with underlying chronic liver disease. Additionally, the tumorous lesions of HCC patients are consistently characterized by the lack of iron accumulation even when arising in iron-loaded liver. However, the molecular mechanism leading to this observed phenomenon is currently poorly understood. In this study, all tumorous tissues from 24 HCC patients with chronic HBV infection were stained negative for iron when histologically assessed by Perls' Prussian blue stain, whereas excess iron deposits were present in 17 of the 24 adjacent non-tumorous liver tissues. To elucidate the concerted regulation of iron homeostasis in these patients, we studied the gene expression profiling of 42 relevant iron-regulatory genes in the tumorous and adjacent non-tumorous liver tissues of these HCC patients along with 10 normal liver controls. Expression for most of the iron-regulatory genes, including hepcidin, transferrin receptor 2 (TfR2), transferrin (Tf), ceruloplasmin (Cp) and iron regulatory protein 1 (IRP1), were significantly down regulated in the tumorous tissues of these patients compared to the adjacent non tumorous liver tissues and normal liver controls. On the other hand, expression of hepcidin, TfR2, ferroportin 1 and DMT1 were significantly up-regulated in iron loaded non-cirrhotic non-tumorous liver tissues as compared with normal liver controls. Hence, the reduction of hepcidin expression within the iron-depleted tumorous lesions likely reflects the physiological consequence of the obligate demand for iron in the rapidly growing neoplastic cells, whereas the up regulation of hepcidin expression in the iron-loaded adjacent non-tumorous liver tissues is likely a physiological response. PMID- 19307464 TI - Altered mechanism of adenosine-induced coronary arteriolar dilation in early stage metabolic syndrome. AB - Onset of the combined metabolic syndrome (MetS) is a complex progressive process involving numerous cardiovascular risk factors. Although patients with established MetS exhibit reduced coronary flow reserve and individual components of the MetS reduce microvascular vasodilation, little is known concerning the impact of early-stage MetS on the mechanisms of coronary flow control. Therefore, we tested the hypothesis that coronary arteriolar dilation to adenosine is attenuated in early-stage MetS by reduced A2 receptor function and diminished K+ channel involvement. Pigs were fed control or high-fat/cholesterol diet for 9 weeks to induce early-stage MetS. Coronary atheroma was determined in vivo with intravascular ultrasound. In vivo coronary dilation was determined by intracoronary adenosine infusion. Further, apical coronary arterioles were isolated, cannulated and pressurized to 60 cmH2O for in vitro pharmacologic assessment of adenosine dilation. Coronary atheroma was not different between groups, indicating early-stage MetS. Coronary arteriolar dilation to adenosine (in vivo) and 2-chloroadenosine (2-CAD; in vitro) was similar between groups. In control arterioles, 2-CAD-mediated dilation was reduced only by selective A(2A) receptor inhibition, whereas only dual A(2A/2B) inhibition reduced this response in MetS arterioles. Arteriolar A(2B), but not A(2A), receptor protein expression was reduced by MetS. Blockade of voltage-dependent K+ (K(v)) channels reduced arteriolar sensitivity to 2-CAD in both groups, whereas ATP-sensitive K+ (K(ATP)) channel inhibition reduced sensitivity only in control arterioles. Our data indicate that the mechanisms mediating coronary arteriolar dilation to adenosine are altered in early-stage MetS prior to overt decrements in coronary vasodilator reserve. PMID- 19307465 TI - Hyperkalemia risk in chronic kidney disease: deterrent to the use of aldosterone receptor antagonism or not. PMID- 19307466 TI - Mechanisms of impaired potassium handling with dual renin-angiotensin-aldosterone blockade in chronic kidney disease. AB - The combination of an aldosterone receptor antagonist added to an angiotensin converting enzyme inhibitor has been demonstrated to reduce cardiovascular and renal end points in hypertensive humans but can produce hyperkalemia in the common clinical setting of impaired renal function. We investigated the effects of dual therapy on acute and chronic potassium handling in hypertensive humans with renal impairment by conducting a randomized crossover clinical trial of 4 weeks of 40 mg lisinopril/25 mg spironolactone versus placebo in 18 participants with a glomerular filtration rate of 25 to 65 mL/min. Study end points, following an established protocol, were hourly determinations of dynamic renal potassium excretion (mmol/h) and serum potassium (mmol/L) after 35 mmol oral potassium challenge in addition to ambulatory potassium concentration. After 4 weeks, ambulatory potassium concentration was 4.87 mmol/L with lisinopril/spironolactone versus 4.37 with placebo (P<0.001). Lisinopril/spironolactone produced only a modest 0.44 mmol/h reduction in stimulated potassium excretion (P=0.03) but a substantial 0.67 mmol/L increase in serum potassium (P<0.001) in response to 35 mmol potassium; these findings are consistent with impaired extrarenal/transcellular potassium disposition. We found the increase in serum potassium after an oral potassium challenge to be a strong predictor of the increase in ambulatory potassium with lisinopril/spironolactone. Our study suggests that dual renin-angiotensin-aldosterone blockade may impair extrarenal/transcellular potassium disposition in addition to reducing potassium excretion in humans with renal impairment, and that acute changes in dynamic potassium handling are predictive of chronic changes in ambulatory potassium concentration with dual renin-angiotensin-aldosterone blockade. PMID- 19307467 TI - Plasma fatty acid composition as a predictor of arterial stiffness and mortality. AB - Aortic stiffness predicts cardiovascular mortality and may be influenced by dietary fat composition. The hypothesis that plasma fat composition influences arterial stiffness and subsequent mortality was tested here in a prospective study. A total of 174 randomly sampled nondiabetic participants aged 45 to 74 years were recruited from local populations, stratified by ethnicity and gender, and followed up for mortality. Aortic pulse wave velocity (PWV), blood pressure, and fatty acid composition of plasma lipids were measured at baseline. PWV was associated with increased cardiovascular mortality and inversely related to the proportions of docosahexaenoic (rho=-0.22; P=0.02) and arachidonic acids (rho= 0.25; P<0.001) in plasma lipids. Principal component analyses identified a cluster characterized by higher proportions of palmitate, palmitoleic and oleic acid and lower proportions of linoleic, dihomo-gamma linolenic, and arachidonic acids. This cluster was positively associated with PWV, central adiposity, smoking, and increased mortality (hazard ratio: 1.13; 95% CI: 1.01 to 1.27). A second cluster, with higher proportions of arachidonic, eicosapentaenoic, and docosahexaenoic and lower proportions of oleic, palmitic, and linoleic acid levels, was associated with lower PWV and systolic blood pressure but also decreased risk of mortality (hazard ratio: 0.57; 95% CI: 0.39 to 0.82), independent of PWV and blood pressure. These data suggest that plasma fatty acid profiles characterized by a higher proportion of long-chain polyunsaturated fatty acids are associated with decreased cardiovascular mortality, independent of the impact of aortic PWV. The results are consistent with an effect of dietary sources of n-3 long-chain polyunsaturates influencing arterial stiffness and mortality. PMID- 19307468 TI - Nanoparticle-mediated drug delivery and pulmonary hypertension. PMID- 19307469 TI - Nanoparticle-mediated delivery of nuclear factor kappaB decoy into lungs ameliorates monocrotaline-induced pulmonary arterial hypertension. AB - Pulmonary arterial hypertension (PAH) is an intractable disease of the small pulmonary artery that involves multiple inflammatory factors. We hypothesized that a redox-sensitive transcription factor, nuclear factor kappaB (NF-kappaB), which regulates important inflammatory cytokines, plays a pivotal role in PAH. We investigated the activity of NF-kappaB in explanted lungs from patients with PAH and in a rat model of PAH. We also examined a nanotechnology-based therapeutic intervention in the rat model. Immunohistochemistry results indicated that the activity of NF-kappaB increased in small pulmonary arterial lesions and alveolar macrophages in lungs from patients with PAH compared with lungs from control patients. In a rat model of monocrotaline-induced PAH, single intratracheal instillation of polymeric nanoparticles (NPs) resulted in delivery of NPs into lungs for or =200 mg/dL, fasting plasma glucose > or =126 mg/dL, or treatment. Among women, the hazard ratios (HRs) for all-cause mortality in the later versus the earlier time period were 0.59 (95% confidence interval, 0.50 to 0.70; P<0.0001) for those without diabetes mellitus and 0.48 (95% confidence interval, 0.32 to 0.71; P=0.002) for those with diabetes mellitus. Similar results were observed in men. Among women and men, the HR of cardiovascular disease mortality declined among those with and without diabetes mellitus. Non-cardiovascular disease mortality declined among women without diabetes mellitus (HR, 0.76; P=0.01), whereas no change was observed among women with diabetes mellitus or among men with or without diabetes mellitus. Individuals with versus those without diabetes mellitus were at increased risk of all-cause mortality in the earlier (HR, 2.44; P<0.0001) and later (HR, 1.95; P<0.0001) time periods. CONCLUSIONS: Reductions in all-cause mortality among women and men with diabetes mellitus have occurred over time. However, mortality rates among individuals with diabetes mellitus remain approximately 2-fold higher compared with individuals without diabetes mellitus. PMID- 19307473 TI - Arterial and aortic valve calcification abolished by elastolytic cathepsin S deficiency in chronic renal disease. AB - BACKGROUND: Clinical studies have demonstrated that 50% of individuals with chronic renal disease (CRD) die of cardiovascular causes, including advanced calcific arterial and valvular disease; however, the mechanisms of accelerated calcification in CRD remain obscure, and no therapies can prevent disease progression. We recently demonstrated in vivo that inflammation triggers cardiovascular calcification. In vitro evidence also indicates that elastin degradation products may promote osteogenesis. Here, we used genetically modified mice and molecular imaging to test the hypothesis in vivo that cathepsin S (catS), a potent elastolytic proteinase, accelerates calcification in atherosclerotic mice with CRD induced by 5/6 nephrectomy. METHODS AND RESULTS: Apolipoprotein-deficient (apoE(-/-))/catS(+/+) (n=24) and apoE(-/-)/catS(-/-) (n=24) mice were assigned to CRD and control groups. CRD mice had significantly higher serum phosphate, creatinine, and cystatin C levels than those without CRD. To visualize catS activity and osteogenesis in vivo, we coadministered catS activatable and calcification-targeted molecular imaging agents 10 weeks after nephrectomy. Imaging coregistered increased catS and osteogenic activities in the CRD apoE(-/-)/catS(+/+) cohort, whereas CRD apoE(-/-)/catS(-/-) mice exhibited less calcification. Quantitative histology demonstrated greater catS-associated elastin fragmentation and calcification in CRD apoE(-/-)/catS(+/+) than CRD apoE( /-)/catS(-/-) aortas and aortic valves. Notably, catS deletion did not cause compensatory increases in RNA levels of other elastolytic cathepsins or matrix metalloproteinases. Elastin peptide and recombinant catS significantly increased calcification in smooth muscle cells in vitro, a process further amplified in phosphate-enriched culture medium. CONCLUSIONS: The present study provides direct in vivo evidence that catS-induced elastolysis accelerates arterial and aortic valve calcification in CRD, providing new insight into the pathophysiology of cardiovascular calcification. PMID- 19307474 TI - Molecular magnetic resonance imaging of myocardial perfusion with EP-3600, a collagen-specific contrast agent: initial feasibility study in a swine model. AB - BACKGROUND: Cardiac magnetic resonance (MR) perfusion imaging during the first pass after intravenous administration of extracellular contrast agents is hampered by the spatial and temporal resolution achievable and by the artifacts seen in ultrafast MR imaging. Furthermore, time-consuming quantitative data analysis is often added. The use of molecular MR imaging with a target-specific contrast agent with perfusion-dependent binding to myocardium may enable prolonged visualization of perfusion defects and thus may help to overcome limitations of currently used first-pass extracellular MR imaging. EP-3600 is a new gadolinium-containing molecular contrast agent that binds reversibly to myocardial collagen. METHODS AND RESULTS: A significant but nonocclusive coronary artery stenosis was modeled in 7 domestic swine with an undersized MR-compatible balloon positioned in the left anterior descending artery as verified by x-ray angiography. Two animals died before contrast injection as a result of arrhythmias. In 5 swine, high-spatial-resolution gradient echo imaging (approximately 1 x 1 mm(2) in-plane resolution) was performed before and 5, 20, 40, and 60 minutes after intravenous administration of 12.3 micromol/kg EP-3600. Contrast was administered during stress induced by an infusion of 250 mumol x kg( 1) x min(-1) adenosine. Yb-DTPA was administered simultaneously for comparison of myocardium-to-plasma ratios. Images were assessed subjectively by 2 investigators, and signal-to-noise and contrast-to-noise ratios over time were calculated. Normal myocardium showed a significant signal-to-noise ratio increase during the entire examination time. In all animals (n=5), the perfusion defect in the left anterior descending artery territory could be visualized with a high contrast-to-noise ratio for at least 20 minutes after contrast injection. A significantly higher myocardium-to-plasma ratio was found for EP-3600 compared with the control agent Yb-DTPA (0.85+/-0.26 versus 0.22+/-0.08, respectively; P<0.01). CONCLUSIONS: EP-3600 is a new molecular MR imaging contrast agent that binds to the myocardium and enables prolonged, high-contrast, high-spatial resolution visualization of myocardial perfusion defects. PMID- 19307475 TI - Metabolomic profiling reveals distinct patterns of myocardial substrate use in humans with coronary artery disease or left ventricular dysfunction during surgical ischemia/reperfusion. AB - BACKGROUND: Human myocardial metabolism has been incompletely characterized in the setting of surgical cardioplegic arrest and ischemia/reperfusion. Furthermore, the effect of preexisting ventricular state on ischemia-induced metabolic derangements has not been established. METHODS AND RESULTS: We applied a mass spectrometry-based platform to profile 63 intermediary metabolites in serial paired peripheral arterial and coronary sinus blood effluents obtained from 37 patients undergoing cardiac surgery, stratified by presence of coronary artery disease and left ventricular dysfunction. The myocardium was a net user of a number of fuel substrates before ischemia, with significant differences between patients with and without coronary artery disease. After reperfusion, significantly lower extraction ratios of most substrates were found, as well as significant release of 2 specific acylcarnitine species, acetylcarnitine and 3 hydroxybutyryl-carnitine. These changes were especially evident in patients with impaired ventricular function, who exhibited profound limitations in extraction of all forms of metabolic fuels. Principal component analysis highlighted several metabolic groupings as potentially important in the postoperative clinical course. CONCLUSIONS: The preexisting ventricular state is associated with significant differences in myocardial fuel uptake at baseline and after ischemia/reperfusion. The dysfunctional ventricle is characterized by global suppression of metabolic fuel uptake and limited myocardial metabolic reserve and flexibility after global ischemia/reperfusion stress in the setting of cardiac surgery. Altered metabolic profiles after ischemia/reperfusion are associated with postoperative hemodynamic course and suggest a role for perioperative metabolic monitoring and targeted optimization in cardiac surgical patients. PMID- 19307476 TI - Associations of gestational weight gain with offspring body mass index and blood pressure at 21 years of age: evidence from a birth cohort study. AB - BACKGROUND: Maternal weight gain in pregnancy is positively associated with offspring body mass index (BMI) and obesity risk in childhood, but whether this increased risk extends into adulthood or results in increases in other cardiovascular risk factors such as elevated blood pressure (BP) is unclear. METHODS AND RESULTS: We used a population-based birth cohort of 2432 individuals (50% male) born in Brisbane, Australia, between 1981 and 1983 to prospectively examine the association between maternal gestational weight gain (GWG) and offspring BMI and BP at 21 years. On average, each mother gained 14.8 kg (SD, 5.1 kg) during her pregnancy. At 21 years of age, offspring mean BMI, systolic BP, and diastolic BP were 24.2 kg/m(2) (SD, 4.9 kg/m(2)), 116.4 mm Hg (SD, 14.5 mm Hg), and 67.7 mm Hg (SD, 8.5 mm Hg), respectively. Offspring BMI was on average 0.3 kg/m(2) (95% confidence interval, 0.1 to 0.4 kg/m(2)) higher for each 0.1 kg/wk greater GWG after adjustment for potential confounding factors. Systolic BP also was greater (0.2 mm Hg per 0.1 kg; 95% confidence interval, -0.2 to 0.6) in offspring whose mothers had higher GWG. Although this association was not statistically significant, it was consistent in magnitude with the association of maternal GWG with offspring BMI and of offspring BMI with BP. CONCLUSIONS: Our findings show that greater GWG is associated with greater offspring BMI into early adulthood and that this may translate into higher systolic BP in offspring. Further large studies are required to confirm an effect of GWG on a range of offspring cardiovascular risk factors. PMID- 19307477 TI - Detection and quantification of left atrial structural remodeling with delayed enhancement magnetic resonance imaging in patients with atrial fibrillation. AB - BACKGROUND: Atrial fibrillation (AF) is associated with diffuse left atrial fibrosis and a reduction in endocardial voltage. These changes are indicators of AF severity and appear to be predictors of treatment outcome. In this study, we report the utility of delayed-enhancement magnetic resonance imaging (DE-MRI) in detecting abnormal atrial tissue before radiofrequency ablation and in predicting procedural outcome. METHODS AND RESULTS: Eighty-one patients presenting for pulmonary vein antrum isolation for treatment of AF underwent 3-dimensional DE MRI of the left atrium before the ablation. Six healthy volunteers also were scanned. DE-MRI images were manually segmented to isolate the left atrium, and custom software was implemented to quantify the spatial extent of delayed enhancement, which was then compared with the regions of low voltage from electroanatomic maps from the pulmonary vein antrum isolation procedure. Patients were assessed for AF recurrence at least 6 months after pulmonary vein antrum isolation, with an average follow-up of 9.6+/-3.7 months (range, 6 to 19 months). On the basis of the extent of preablation enhancement, 43 patients were classified as having minimal enhancement (average enhancement, 8.0+/-4.2%), 30 as having moderate enhancement (21.3+/-5.8%), and 8 as having extensive enhancement (50.1+/-15.4%). The rate of AF recurrence was 6 patients (14.0%) with minimal enhancement, 13 (43.3%) with moderate enhancement, and 6 (75%) with extensive enhancement (P<0.001). CONCLUSIONS: DE-MRI provides a noninvasive means of assessing left atrial myocardial tissue in patients suffering from AF and might provide insight into the progress of the disease. Preablation DE-MRI holds promise for predicting responders to AF ablation and may provide a metric of overall disease progression. PMID- 19307478 TI - Macrophage apoptosis exerts divergent effects on atherogenesis as a function of lesion stage. AB - BACKGROUND: Because apoptotic cell clearance appears to be defective in advanced compared with early atherosclerotic plaques, macrophage apoptosis may differentially affect plaque progression as a function of lesion stage. METHODS AND RESULTS: We first evaluated the impact of targeted protection of macrophages against apoptosis at both early and advanced stages of atherosclerosis. Increased resistance of macrophages to apoptosis in early atherosclerotic lesions was associated with increased plaque burden; in contrast, it afforded protection against progression to advanced lesions. Conversely, sustained induction of apoptosis in lesional macrophages of advanced lesions resulted in a significant increase in lesion size. Such enhanced lesion size occurred as a result not only of apoptotic cell accumulation but also of elevated chemokine expression and subsequent intimal recruitment of circulating monocytes. CONCLUSIONS: Considered together, our data suggest that macrophage apoptosis is atheroprotective in fatty streak lesions, but in contrast, defective clearance of apoptotic debris in advanced lesions favors arterial wall inflammation and enhanced recruitment of monocytes, leading to enhanced atherogenesis. PMID- 19307479 TI - Stress Doppler echocardiography in relatives of patients with idiopathic and familial pulmonary arterial hypertension: results of a multicenter European analysis of pulmonary artery pressure response to exercise and hypoxia. AB - BACKGROUND: This large, prospective, multicentric study was performed to analyze the distribution of tricuspid regurgitation velocity (TRV) values during exercise and hypoxia in relatives of patients with idiopathic and familial pulmonary arterial hypertension (PAH) and in healthy control subjects. We tested the hypothesis that relatives of idiopathic/familial PAH patients display an enhanced frequency of hypertensive TRV response to stress and that this response is associated with mutations in the bone morphogenetic protein receptor II (BMPR2) gene. METHODS AND RESULTS: TRV was estimated by Doppler echocardiography during supine bicycle exercise in normoxia and during 120 minutes of normobaric hypoxia (FIO(2)=12%; approximately 4500 m) in 291 relatives of 109 PAH patients and in 191 age-matched control subjects. Mean maximal TRVs were significantly higher in PAH relatives during both exercise and hypoxia. During exercise, 10% of control subjects but 31.6% of relatives (P<0.0001) exceeded the 90% quantile of mean maximal TRV seen in control subjects. Hypoxia revealed hypertensive TRV in 26% of relatives (P=0.0029). Among control subjects, TRV at rest was not related to age, sex, body mass index, systemic blood pressure, smoking status, or heart rate. Within kindreds identified as harboring deleterious mutations of the BMPR2 gene, a hypertensive TRV response occurred significantly more often compared with those without detected mutations. CONCLUSIONS: Pulmonary hypertensive response to exercise and hypoxia in idiopathic/familial PAH relatives appears as a genetic trait with familial clustering, being correlated to but not caused by a BMPR2 mutation. The suitability of this trait to predict manifest PAH development should be addressed in long-term follow-up studies. PMID- 19307480 TI - Collagen-targeting vascular endothelial growth factor improves cardiac performance after myocardial infarction. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is an important active protein for the induction of angiogenesis and improvement in cardiac function after myocardial ischemia; however, the lack of a delivery system targeted to the injured myocardium reduces the local therapeutic efficacy of VEGF and increases its possible adverse effects. METHODS AND RESULTS: We produced a fusion protein (CBD-VEGF) consisting of VEGF and a collagen-binding domain (CBD). The fusion protein specifically bound to type I collagen in vitro. In addition, CBD-VEGF promoted human umbilical vein endothelial cell proliferation after binding to collagen, which indicates that it retained both growth factor activity and collagen-binding ability. When implanted subcutaneously in rats, collagen membranes loaded with CBD-VEGF were significantly vascularized. After it was injected into rats with acute myocardial infarction, CBD-VEGF was largely retained in the cardiac extracellular matrix, in which collagen I was rich. Four weeks after VEGF or CBD-VEGF was injected into the infarct border zone, cardiac function detected by echocardiography and hemodynamics was preserved in the CBD VEGF group. Administration of CBD-VEGF also induced reduction of scar size, whereas native VEGF did not have these effects. In addition, a significant increase in the number of capillary vessels in infarcted hearts was found in the CBD-VEGF group. CONCLUSIONS: The injection of CBD-VEGF improved cardiac function in rats with induced acute myocardial infarction. This could potentially provide a new treatment option for myocardial infarction. PMID- 19307481 TI - Syncope and risk of sudden death in hypertrophic cardiomyopathy. AB - BACKGROUND: The prognostic significance of syncope has not been investigated systematically in hypertrophic cardiomyopathy, and treatment strategies have been based largely on intuition and experience. METHODS AND RESULTS: We assessed the relationship between syncope and sudden death in 1511 consecutive patients with hypertrophic cardiomyopathy. Unexplained (n=153) or neurally mediated (n=52) syncope occurred in 205 patients (14%). Over a 5.6+/-5.2-year follow-up, 74 patients died suddenly. Relative risk of sudden death was 1.78 (95% confidence interval 0.88 to 3.51, P=0.08) in patients with unexplained syncope and 0.91 (95% confidence interval 0.00 to 3.83, P=1.0) in those with neurally mediated syncope compared with patients without syncope. In multivariable analysis, the temporal proximity of unexplained syncope to initial patient evaluation was independently associated with risk of sudden death (P=0.006). Patients with unexplained syncope within 6 months before the initial evaluation showed a 5-fold increase in risk compared with patients without syncope (adjusted hazard ratio 4.89, 95% confidence interval 2.19 to 10.94), a relationship that was maintained throughout all age groups (<18, 18 to 39, and > or =40 years). Older patients (> or =40 years of age) with remote episodes of syncope (>5 years before initial evaluation) did not show an increased risk of sudden death (adjusted hazard ratio 0.38, 95% confidence interval 0.05 to 2.74). CONCLUSIONS: In the present large cohort of patients with hypertrophic cardiomyopathy, unexplained syncope was a risk factor for sudden death. Patients with syncopal events that occurred in close temporal proximity to the initial evaluation showed a substantially higher risk of sudden death than patients without syncope. Older patients with remote syncopal events did not show an increased risk. PMID- 19307482 TI - Images in cardiovascular medicine. Intralobar pulmonary sequestration in a 46 year-old woman: findings from multidetector-row computed tomography and magnetic resonance imaging. PMID- 19307483 TI - Re: Trimetazidine, a metabolic modulator, has cardiac and extracardiac benefits in idiopathic dilated cardiomyopathy. PMID- 19307485 TI - Discarding logic: 2008 Ancel Keys Memorial Lecture. PMID- 19307486 TI - Images in cardiovascular medicine. Posttraumatic cardiac contrecoup: in vivo evidence by cardiac magnetic resonance imaging. PMID- 19307488 TI - Prophylactic intravenous immunoglobulin does not have a role in hematopoietic stem-cell transplantation: is the evidence clear? PMID- 19307489 TI - Comprehensive screening for Lynch syndrome: who can be the driving force in daily clinical practice? PMID- 19307491 TI - Every cancer patient needs radiation therapy! PMID- 19307492 TI - Cancer during pregnancy: the time has come for a prospective program. PMID- 19307493 TI - Disparities in breast cancer adjuvant chemotherapy: moving beyond yes or no. PMID- 19307494 TI - Vox populi: Using community-based studies to determine best management of early stage nonseminoma. PMID- 19307496 TI - Does bortezomib induce de facto varicella zoster virus reactivation in patients with multiple myeloma? PMID- 19307497 TI - Heeding our words: complexities of research among low-literacy populations. PMID- 19307498 TI - Protein-losing enteropathy due to T-cell large granular lymphocyte leukemia. PMID- 19307499 TI - Implications of familial colorectal cancer risk profiles and microsatellite instability status. AB - PURPOSE: Estimating familial colorectal cancer (CRC) risk is clinically important in being able to discriminate between high- and low-risk groups. To quantify familial CRC risks associated with mismatch repair (MMR) deficient and microsatellite stable (MSS) tumors, we analyzed 2,941 population-based cases of CRC. PATIENTS AND METHODS: MMR status in CRCs was established by testing for microsatellite instability (MSI). MUTYH status was assigned by screening for Y165C and G382D variants. Age-specific relative and absolute CRC risks in first degree relatives (FDRs) were calculated, and the most likely genetic models of familial aggregation were derived. RESULTS: CRC risks in FDRs were strongly associated with MSI status (MSI, standardized incidence ratio [SIR] = 4.28, 95% CI, 3.51 to 5.17; MSS, SIR = 1.91, 95% CI, 1.73 to 2.11), early-onset disease (MSI patient age < 55 years, SIR = 10.96, 95% CI, 8.32 to 14.17; MSS patient age < 55 years, SIR = 2.3, 95% CI, 1.88 to 2.85), and having more than one affected FDR (MSI, SIR = 10.00, 95% CI, 7.74 to 12.72; MSS, SIR = 2.78, 95% CI, 2.18 to 3.48). The familial aggregation of CRC associated with MSI cancer was parsimonious with dominant model conferring a high CRC risk at early ages. Approximately 69% of the excess familial risk in FDRs can be ascribed to MSS CRC, and although the pattern of familial risk supports recessive susceptibility in addition to MUTYH, the absolute risk of CRC is at best modest. CONCLUSION: The results from this analysis should enable an individual's risk of CRC to be more accurately estimated, thus maximizing the value of screening programs. Results also have utility in the design of genetic analyses to identify novel disease alleles. PMID- 19307500 TI - Phase III trial of bevacizumab in combination with gemcitabine and erlotinib in patients with metastatic pancreatic cancer. AB - PURPOSE: Treatment with gemcitabine provides modest benefits in patients with metastatic pancreatic cancer. The addition of erlotinib to gemcitabine shows a small but significant improvement in overall survival (OS) versus gemcitabine alone. Phase II results for bevacizumab plus gemcitabine provided the rationale for a phase III trial of gemcitabine-erlotinib plus bevacizumab or placebo. PATIENTS AND METHODS: Patients with metastatic pancreatic adenocarcinoma were randomly assigned to receive gemcitabine (1,000 mg/m(2)/week), erlotinib (100 mg/day), and bevacizumab (5 mg/kg every 2 weeks) or gemcitabine, erlotinib, and placebo in this double-blind, phase III trial. Primary end point was OS; secondary end points included progression-free survival (PFS), disease control rate, and safety. RESULTS: A total of 301 patients were randomly assigned to the placebo group and 306 to the bevacizumab group. Median OS was 7.1 and 6.0 months in the bevacizumab and placebo arms, respectively (hazard ratio [HR], 0.89; 95% CI, 0.74 to 1.07; P = .2087); this difference was not statistically significant. Adding bevacizumab to gemcitabine-erlotinib significantly improved PFS (HR, 0.73; 95% CI, 0.61 to 0.86; P = .0002). Treatment with bevacizumab plus gemcitabine erlotinib was well tolerated: safety data did not differ from previously described safety profiles for individual drugs. CONCLUSION: The primary objective was not met. The addition of bevacizumab to gemcitabine-erlotinib did not lead to a statistically significant improvement in OS in patients with metastatic pancreatic cancer. PFS, however, was significantly longer in the bevacizumab group compared with placebo. No unexpected safety events were observed from adding bevacizumab to gemcitabine-erlotinib. PMID- 19307501 TI - Chemoradiotherapy in the management of locally advanced pancreatic carcinoma: a qualitative systematic review. AB - PURPOSE: Pancreatic carcinoma is one of the leading causes of cancer-related mortality. At time of diagnosis, 30% of patients present with a locally advanced unresectable but nonmetastatic pancreatic carcinoma (LAPC). The French program Standards, Options, and Recommendations was promoted to conduct a qualitative systematic review to evaluate the role of radiotherapy in patients with LAPC. METHODS: A search to identify eligible studies was undertaken using the MEDLINE database. All phase III randomized trials and systematic reviews evaluating the role of radiotherapy in LAPC were included, together with some noncontrolled studies if no phase III trials were retrieved. The quality and clinical relevance of the studies were evaluated using validated checklists, which allowed associating each result with a level of evidence. RESULTS: Twenty-one studies were included, as follows: two meta-analyses, 13 randomized trials, and six nonrandomized trials. Chemoradiotherapy increases overall survival when compared with best supportive care (level of evidence C) or with exclusive radiotherapy (level B1), but is more toxic (level B1). Chemoradiotherapy is not superior to chemotherapy in terms of survival (level B1) and increases toxicity (level A). Recent data favor limited irradiation to the tumor volume (level C). Fluorouracil is still the reference chemotherapy in association with radiotherapy (level B1). Induction chemotherapy before chemoradiotherapy improves survival (level C). CONCLUSION: No standard treatment exists, but there are two options for treatment of LAPC; these are gemcitabine-based chemotherapy and chemoradiotherapy. Induction chemotherapy followed by a chemoradiotherapy is a promising strategy for selection of patients without early metastatic/progressing disease. PMID- 19307502 TI - Overcoming resistance to conventional drugs in Ewing sarcoma and identification of molecular predictors of outcome. AB - PURPOSE: The improvement of Ewing sarcoma (EWS) therapy is currently linked to the discovery of strategies to select patients with poor and good prognosis and of modified treatment regimens. In this study, we analyzed the molecular factors governing EWS response to chemotherapy to identify genetic signatures to be used for risk-adapted therapy. PATIENTS AND METHODS: Microarray technology was used for profiling 30 primary tumors and seven metastases of patients who were classified according to event-free survival. For selected genes, real-time polymerase chain reaction was applied in 42 EWS primary tumors as validation assay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test was used to evaluate in vitro drug sensitivity. RESULTS: We identified molecular signatures that reflect tumor resistance to chemotherapy. Annotation analysis was applied to reveal the biologic functions that critically influenced clinical outcome. The prognostic relevance of glutathione metabolism pathway was validated. The expression of MGST1, the microsomal glutathione S-transferase (GST), was found to clearly predict EWS prognosis. MGST1 expression was associated with doxorubicin chemosensitivity. This prompted us to assess the in vitro effectiveness of 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthio)hexanol (NBDHEX), a new anticancer agent that efficiently inhibits GST enzymes. Six cell lines were found to be sensitive to this new drug. CONCLUSION: Classification of EWS patients into high- and low-risk groups is feasible with restricted molecular signatures that may have practical value at diagnosis for selecting patients with EWS who are unresponsive to current treatments. Glutathione metabolism pathway emerged as one of the most significantly altered prognosis-associated pathway. NBDHEX is proposed as a new potential therapeutic possibility. PMID- 19307503 TI - Randomized phase II and pharmacogenetic study of pemetrexed compared with pemetrexed plus carboplatin in pretreated patients with advanced non-small-cell lung cancer. AB - PURPOSE: We performed a randomized phase II trial comparing pemetrexed with pemetrexed plus carboplatin (PC) in patients experiencing relapse after platinum based chemotherapy. PATIENTS AND METHODS: Main eligibility criteria were histologic or cytologic proof of advanced non-small-cell lung cancer (NSCLC), relapse more than 3 months after platinum-based chemotherapy, normal organ function, and Eastern Cooperative Oncology Group performance status 0 to 2. Patients were randomly assigned to pemetrexed 500 mg/m(2) (arm A) or carboplatin area under the curve 5 and pemetrexed 500 mg/m(2) (arm B), both administered intravenously every 3 weeks. Response assessment was performed every 6 weeks; toxicity assessment was performed every 3 weeks. Primary end point was time to progression (TTP); secondary end points were objective response rate (ORR), overall survival (OS), and toxicity. The study was designed to detect a 33% decrease in the hazard of disease progression in the combination arm (alpha = 0.05, two-sided log-rank test). Polymorphisms of thymidylate synthase, the reduced folate carrier, gamma-glutamyl hydrolase, and methylenetetrahydrofolate reductase (MTHF) were investigated in peripheral WBCs of consenting patients. RESULTS: Two hundred forty patients were enrolled. Median TTP was 2.8 months for arm A versus 4.2 months for arm B (hazard ratio, 0.67; 95% CI, 0.51 to 0.89; P = .005). Median OS was 7.6 months and 8.0 months and ORR was 4% and 9% for arms A and B, respectively. Subgroup analyses found adenocarcinoma to be associated with favorable outcome. Toxicities in both arms was negligible, with one potential toxic death in arm A. Patients with MTHFR C677T homozygous mutation had increased progression-free survival compared with patients with wild-type or heterozygous mutations (P = .03). CONCLUSION: PC as second-line treatment for relapsed NSCLC resulted in a significant 33% reduction of the hazard of disease progression as compared with pemetrexed alone. PMID- 19307504 TI - Treatment quality and outcomes of African American versus white breast cancer patients: retrospective analysis of Southwest Oncology studies S8814/S8897. AB - PURPOSE: Women of African ancestry (AA) have lower WBC counts and are more likely to have treatment delays and discontinue adjuvant breast cancer therapy early compared with white women. We assessed the association between race and treatment discontinuation/delay, WBC counts, and survival in women enrolled onto breast cancer clinical trials. PATIENTS AND METHODS: AA and white women from Southwest Oncology Group adjuvant breast cancer trials (S8814/S8897) were matched by age and protocol. Only the treatment arms in which patients were scheduled to receive six cycles of chemotherapy were analyzed. RESULTS: A total of 317 pairs of patients (n = 634) were analyzed. At baseline, AA women had higher body-surface area (P < .0001) and lower WBC (P = .0009). AA women were more likely to have tumors that were > or = 2 cm (P = .01) and hormone receptor negative (P < .0001). AA women, versus white women, were marginally more likely to discontinue treatment early (11% v 7%, respectively; P = .07) or have one or more treatment delays (85% v 79%, respectively; P = .07) and were significantly more likely to experience the combined end point (discontinuation/delay; 87% v 81%, respectively; P = .04). The mean relative dose-intensity (RDI) was similar for both groups (87% in AA women v 86% in white women); however, overall, 43% had an RDI of less than 85%. After adjusting for baseline WBC and prognostic factors in a multivariate model, AA women had worse disease-free survival (hazard ratio [HR] = 1.56; 95% CI, 1.15 to 2.11; P = .005) and overall survival (HR = 1.95; 95% CI, 1.36 to 2.78; P = .0002). The inclusion of RDI and treatment delivery/quality in the regression had little impact on the results. CONCLUSION: On cooperative group breast cancer trials, AA and white women had similar RDIs, but AA women were more likely to experience early discontinuation or treatment delay. Despite correcting for these factors and known predictors of outcome, AA women still had worse survival. PMID- 19307505 TI - Phase II trial of vorinostat in recurrent glioblastoma multiforme: a north central cancer treatment group study. AB - PURPOSE: Vorinostat, a histone deacetylase inhibitor, represents a rational therapeutic target in glioblastoma multiforme (GBM). PATIENTS AND METHODS: Patients with recurrent GBM who had received one or fewer chemotherapy regimens for progressive disease were eligible. Vorinostat was administered at a dose of 200 mg orally twice a day for 14 days, followed by a 7-day rest period. RESULTS: A total of 66 patients were treated. Grade 3 or worse nonhematologic toxicity occurred in 26% of patients and consisted mainly of fatigue (17%), dehydration (6%), and hypernatremia (5%); grade 3 or worse hematologic toxicity occurred in 26% of patients and consisted mainly of thrombocytopenia (22%). Pharmacokinetic analysis showed lower vorinostat maximum concentration and area under the curve (0 to 24 hours) values in patients treated with enzyme-inducing anticonvulsants, although this did not reach statistical significance. The trial met the prospectively defined primary efficacy end point, with nine of the first 52 patients being progression-free at 6 months. Median overall survival from study entry was 5.7 months (range, 0.7 to 28+ months). Immunohistochemical analysis performed in paired baseline and post-vorinostat treatment samples in a separate surgical subgroup of five patients with recurrent GBM showed post treatment increase in acetylation of histones H2B and H4 (four of five patients) and of histone H3 (three of five patients). Microarray RNA analysis in the same samples showed changes in genes regulated by vorinostat, such as upregulation of E cadherin (P = .02). CONCLUSION: Vorinostat monotherapy is well tolerated in patients with recurrent GBM and has modest single-agent activity. Histone acetylation analysis and RNA expression profiling indicate that vorinostat in this dose and schedule affects target pathways in GBM. Additional testing of vorinostat in combination regimens is warranted. PMID- 19307506 TI - Risk-adapted treatment in clinical stage I nonseminomatous germ cell testicular cancer: the SWENOTECA management program. AB - PURPOSE: To offer minimized risk-adapted adjuvant treatment on a nationwide basis for patients with clinical stage 1 (CS1) nonseminomatous germ-cell testicular cancer (NSGCT). The aim was to reduce the risk of relapse and thereby reducing the need of later salvage chemotherapy while maintaining a high cure rate. PATIENTS AND METHODS: From 1998 to 2005, 745 Norwegian and Swedish patients were included into a prospective, community-based multicenter Swedish and Norwegian Testicular Cancer Project (SWENOTECA) management program. Treatment strategy depended on the presence or absence of vascular tumor invasion (VASC). VASC positive patients were recommended brief adjuvant chemotherapy (ACT) with bleomycin, etoposide, and cisplatin (BEP), whereas VASC-negative patients could choose between ACT and surveillance. RESULTS: At a median follow-up of 4.7 years, there have been 51 relapses. On surveillance, 41.7% of VASC+ patients relapsed, compared with 13.2% of VASC- patients. After one course of BEP, 3.2% of VASC+ and 1.3% of VASC- patients relapsed. The toxicity of adjuvant BEP was low. Eight patients have died, none died from progressive disease. CONCLUSION: One course of adjuvant BEP reduces the risk of relapse by approximately 90% in both VASC+ and VASC- CS1 NSGCT, and may be a new option as initial treatment for all CS1 NSGCT. One course of adjuvant BEP for VASC+ CS1 reduces the total burden of chemotherapy compared with surveillance or two courses of BEP. SWENOTECA currently recommends one course of BEP as standard treatment of VASC+ CS1 NSGCT, whereas both surveillance and one course of BEP are options for VASC- CS1 NSGCT. PMID- 19307507 TI - Serum amyloid A as a prognostic marker in melanoma identified by proteomic profiling. AB - PURPOSE: Currently known prognostic serum biomarkers of melanoma are powerful in metastatic disease, but weak in early-stage patients. This study was aimed to identify new prognostic biomarkers of melanoma by serum mass spectrometry (MS) proteomic profiling, and to validate candidates compared with established markers. PATIENTS AND METHODS: Two independent sets of serum samples from 596 melanoma patients were investigated. The first set (stage I = 102; stage IV = 95) was analyzed by matrix assisted laser desorption and ionization time of flight (MALDI TOF) MS for biomarkers differentiating between stage I and IV. In the second set (stage I = 98; stage II = 91; stage III = 87; stage IV = 103), the serum concentrations of the candidate marker serum amyloid A (SAA) and the known biomarkers S100B, lactate dehydrogenase, and C reactive protein (CRP) were measured using immunoassays. RESULTS: MALDI TOF MS revealed a peak at m/z 11.680 differentiating between stage I and IV, which could be identified as SAA. High peak intensities at m/z 11.680 correlated with poor survival. In univariate analysis, SAA was a strong prognostic marker in stage I to III (P = .043) and stage IV (P = .000083) patients. Combination of SAA and CRP increased the prognostic impact to P = .011 in early-stage (I to III) patients. Multivariate analysis revealed sex, stage, tumor load, S100B, SAA, and CRP as independent prognostic factors, with an interaction between SAA and CRP. In stage I to III patients, SAA combined with CRP was superior to S100B in predicting patients' progression-free and overall survival. CONCLUSION: SAA combined with CRP might be used as prognostic serological biomarkers in early-stage melanoma patients, helping to discriminate low-risk patients from high-risk patients needing adjuvant treatment. PMID- 19307508 TI - Patient involvement in decisions to limit treatment: the crucial role of agreement between physician and patient. AB - PURPOSE: The aim of this study was to describe, first, the decision-making process concerning the limitation of life-prolonging treatment (DLT); second, the extent to which patients are actually involved in these decisions; and third, to detect medical and ethical factors that affect patient involvement. PATIENTS AND METHODS: This prospective qualitative study enrolled 76 patients with incurable cancer with whom the limitation of life-prolonging treatment was discussed. Embedded researchers on the wards recorded the patient's history, medical condition, type of treatment limitation discussed, patient wishes, decision making capacity, and patient involvement using an in-depth documentation procedure. RESULTS: While the majority of patients were informed about their diagnosis, therapy, and course of disease (99%, 97%, 90%, respectively), only 47% were involved in DLT. Two thirds of the patients preferred palliative care, and one third wished to extend their lifetime. If patients preferred palliative care, they were more often in line with physicians' treatment goals than patients who were striving for longer survival (91.4% v 46.7%; P = .001). They also were involved significantly more often in DLT. Multivariate analysis showed that age, Karnofsky performance index or decision-making capacity had no impact on patient involvement. CONCLUSION: Only half of the patients were involved in DLT. Surprisingly, the main predictor of patient involvement was not their medical condition, but agreement with physicians' palliative treatment goals. These results show that if physicians switch to comfort care in terminally ill patients and patients are not yet prepared to follow this line, treatment limitations are often decided without involving the patient. PMID- 19307509 TI - Impact of diabetes mellitus on complications and outcomes of adjuvant chemotherapy in older patients with breast cancer. AB - PURPOSE: To evaluate whether diabetes affects patterns of adjuvant chemotherapy use, toxic effects of chemotherapy, and breast cancer outcomes. PATIENTS AND METHODS: By using the Surveillance, Epidemiology, and End Results-Medicare database, we identified patients aged 66 years or older who had stages I through III breast cancer that was diagnosed between 1992 and 2002. Multivariable regression analyses were performed to determine the effect of diabetes on use of chemotherapy, toxicities, and outcomes. The risks of all-cause mortality and breast cancer-specific (BCS) mortality were estimated with the Kaplan-Meier method. RESULTS: Our cohort had 70,781 men and women, of whom 14,414 (20.36%) had diabetes. Among people who received chemotherapy (n = 11,826), 21.0% were diabetics. In this group, diabetics had lower odds of receiving anthracyclines (odds ratio [OR], 0.78; 95% CI, 0.71 to 0.87) and taxanes (OR, 0.86; 95% CI, 0.75 to 0.99). Diabetes was associated with increased odds of being hospitalized for any chemotherapy toxicity (OR, 1.38; 95% CI, 1.23 to 1.56), for infection or fever (OR, 1.43; 95% CI, 1.2 to 1.7), for neutropenia (OR, 1.22; 95% CI, 1.03 to 1.45), for anemia (OR, 1.24; 95% CI, 1.05 to 1.47), and for any cause (OR, 1.32; 95% CI, 1.19 to 1.46). Patients with diabetes had higher all-cause mortality (hazard ratio [HR], 1.35; 95% CI, 1.31 to 1.39). There was a significant interaction between diabetes and chemotherapy use for BCS mortality. Diabetic and nondiabetic patients who did not receive chemotherapy had similar BCS mortality, but diabetic patients who did receive chemotherapy had higher BCS mortality than nondiabetic patients (OR, 1.20; 95% CI, 1.07 to 1.35). CONCLUSION: In this observational, hypothesis-generating study, patients who have breast cancer and diabetes are at increased risk of chemotherapy-related toxicities compared with nondiabetic patients who are receiving chemotherapy and have higher all-cause mortality. PMID- 19307510 TI - Phase II studies of gemcitabine and cisplatin in heavily and minimally pretreated metastatic breast cancer. AB - PURPOSE: Cisplatin and gemcitabine have single-agent activity in metastatic breast cancer, and preclinical data support synergy of the combination. Two parallel, phase II trials were conducted to evaluate the response rate, response duration, and toxicities of the combination. Genetic polymorphisms were analyzed for correlation with outcomes. PATIENTS AND METHODS: Eligible women had measurable disease and heavily or minimally pretreated metastatic breast cancer. The heavily pretreated protocol required prior anthracycline and taxane therapy; cisplatin as part of high-dose therapy was allowed. All patients received cisplatin 25 mg/m(2) on days 1 through 4 and gemcitabine 1,000 mg/m(2) on days 2 and 8 of a 21-day cycle with prophylactic granulocyte colony-stimulating factor in the heavily pretreated group. Sera from a subset of patients were evaluated by polymerase chain reaction restriction fragment length polymorphism for polymorphisms in 10 genes of interest. RESULTS: Of 136 women enrolled, 74 were heavily pretreated. Both protocols accrued to their two-stage design. The response rate for both the heavily and minimally pretreated cohorts was 26%, and the median durations of response were 5.3 and 5.9 months, respectively. In a multivariate analysis, hormone receptor-negative disease was associated with a higher response rate. The most common grades 3 or 4 toxicities were thrombocytopenia (71%), neutropenia (66%), and anemia (38%). In a subset of 55 patients, the xeroderma pigmentosum group D (XPD)-751, x-ray cross-complementing group 3 (XRCC3) and cytidine deaminase polymorphisms were significantly associated with clinical outcomes. CONCLUSION: Combination cisplatin and gemcitabine is active in metastatic breast cancer regardless of prior therapy. Genetic polymorphisms may tailor which patients benefit from this regimen. PMID- 19307511 TI - Impact of the duration of adjuvant hormonal therapy in patients with locally advanced prostate cancer treated with radiotherapy: a secondary analysis of RTOG 85-31. AB - PURPOSE: Radiation Therapy Oncology Group 85-31 was a randomized trial of androgen suppression for life for patients with locally advanced prostate cancer. However, not all patients continued on the protocol-mandated long-term hormonal therapy despite no evidence of recurrence. We correlated duration of adjuvant hormonal therapy and outcomes among patients who prematurely discontinued hormonal therapy. PATIENTS AND METHODS: The protocol mandated pelvic radiotherapy followed by goserelin given indefinitely or until disease progression. There were 189 analyzable patients. Patients were divided in groups based on the tertile of hormonal therapy duration (HTD) as follows: < or = 1 year, more than 1 year and < or = 5 years, and more than 5 years. Overall survival (OS), disease-free survival (DFS), cause-specific mortality, local failure (LF), and distant metastasis (DM) were studied. Kaplan-Meier estimation and Cox proportional hazards regression model were used for OS and DFS, and Fine and Gray's regression model was used for the other outcomes. RESULTS: The median follow-up for surviving patients is 9.6 years. The median duration of adjuvant hormonal therapy was 2.2 years. The HTD more than 5 years group is significantly associated with an improved survival and DFS and fewer DMs than other HTD groups. After adjustment for age, radical prostatectomy, nodal status, Gleason score, and stage variables, the HTD more than 5 years group remains significantly associated with better OS and DFS than other HTD groups. CONCLUSION: In this hypothesis-generating analysis, prolonged HTD of more than 5 years seems significantly associated with improvements in most outcomes. Given these data, decreasing HTD to < or = 5 years may have a detrimental effect on patients with locally advanced prostate cancer. Only a randomized trial will conclusively clarify this issue. PMID- 19307512 TI - Lack of comprehension of common prostate cancer terms in an underserved population. AB - PURPOSE: To assess the comprehension of common medical terms used in prostate cancer in patient education materials to obtain informed consent, and to measure outcomes after prostate cancer treatment. We address this issue among underserved, African-American men because of the increased cancer incidence and mortality observed in this population. PATIENTS AND METHODS: We reviewed patient education materials and prostate-specific quality-of-life instruments to identify technical terms describing sexual, urinary, and bowel function. Understanding of these terms was assessed in face-to-face interviews of 105, mostly African American men, age > or = 40, from two low-income clinics. Comprehension was evaluated using semiqualitative methods coded by two independent investigators. Demographics were collected and literacy was measured. RESULTS: Fewer than 50% of patients understood the terms "erection" or "impotent." Only 5% of patients understood the term "incontinence" and 25% understood the term "bowel habits." More patients recognized word roots than related terms or compound words (eg, "rectum" v "rectal urgency," "intercourse" v "vaginal intercourse"). Comprehension of terms from all domains was statistically significantly correlated with reading level (P < .001). Median literacy level was fourth to sixth grade. Prostate cancer knowledge was poor. Many patients had difficulty locating key anatomic structures. CONCLUSION: Limited comprehension of prostate cancer terms and low literacy create barriers to obtaining informed consent for treatment and to measuring prostate cancer outcomes accurately in our study population. In addition, the level of prostate cancer knowledge was poor. These results highlight the need for prostate cancer education efforts and outcomes measurements that consider literacy and use nonmedical language. PMID- 19307513 TI - Interpreting trial results in light of conflicting evidence: a Bayesian analysis of adjuvant chemotherapy for non-small-cell lung cancer. AB - PURPOSE: When successive randomized trials contradict prior evidence, clinicians may be unsure how to evaluate them: Does accumulating evidence warrant changing practice? An increasingly popular solution, Bayesian statistics quantitatively evaluate new results in context. This study provides a clinically relevant example of Bayesian methods. METHODS: Three recent non-small-cell lung cancer adjuvant chemotherapy trials were evaluated in light of prior conflicting data. Results were used from International Adjuvant Lung Trial (IALT), JBR.10, and Adjuvant Navelbine International Trialist Association (ANITA). Prior evidence was sequentially updated to calculate the probability of each survival benefit level (overall and by stage) and variance. Sensitivity analysis was performed using expert opinion and uninformed estimates of survival benefit prior probability. RESULTS: The probability of a 4% survival benefit increased from 33% before IALT to 64% after IALT. After sequential updating with JBR.10 and ANITA, this probability was 82% (hazard ratio = 0.84; 95% CI, 0.77 to 0.91). IALT produced the largest decrease in variance (61%) and decreased the chance of survival decrement to 0%. Sensitivity analysis did not support a survival benefit after IALT. However, sequential updating substantiated a 4% survival benefit and, for stage II and III, more than 90% probability of a 6% benefit and 50% probability of a 12% benefit. CONCLUSION: When evaluated in context with prior data, IALT did not support a 4% survival benefit. However, sequential updating with JBR.10 and ANITA did. A model for future assessments, this study demonstrates the unique ability of Bayesian analysis to evaluate results that contradict prior evidence. PMID- 19307514 TI - Prospective study of factors predicting adherence to medical advice in men with testicular cancer. AB - PURPOSE: To identify predictive factors of adherence to medical advice, specifically the likelihood of attendance to a recommended follow-up regimen in patients with newly diagnosed testicular cancer. PATIENTS AND METHODS; This was a prospective study measuring initially not only aspects of the doctor-patient interview, but also a range of demographic, psychological, social, and medical factors, and then recording attendance behavior on follow-up. All 209 new patients with testicular cancer referred between June 1992 and May 1995 were approached, and 184 men consented and completed questionnaires. The nonadherence end point (nonattender) was two failures to attend an outpatient appointment at least 1 month apart, despite a written reminder. RESULTS: Thirty-two participants (17%) were classified as nonattenders. No significant differences were found between attenders and nonattenders in the majority of psychosocial and medical variables that might have predicted nonadherence to medical advice. There was a highly significant association between nonattendance and a patient's perception of an unsatisfactory affective relationship with his clinician (P = .005; hazard ratio, 3.1; 95% CI, 1.4 to 6.6). CONCLUSION: Patients who perceived an unsatisfactory affective relationship with their clinician that included an inability to trust the clinician and a perception that they were not being treated as "a person" were subsequently more likely to disregard medical advice regarding follow-up. Attention to the ways young men may wish to communicate with their clinicians is important, bearing in mind that they may not necessarily adhere to stereotypical images of masculine self-dependence. PMID- 19307515 TI - Reducing meat consumption has multiple benefits for the world's health. PMID- 19307516 TI - Myths and misconceptions about {alpha}1-antitrypsin deficiency. PMID- 19307517 TI - Prevention of nonvertebral fractures with oral vitamin D and dose dependency: a meta-analysis of randomized controlled trials. AB - BACKGROUND: Antifracture efficacy with supplemental vitamin D has been questioned by recent trials. METHODS: We performed a meta-analysis on the efficacy of oral supplemental vitamin D in preventing nonvertebral and hip fractures among older individuals (> or =65 years). We included 12 double-blind randomized controlled trials (RCTs) for nonvertebral fractures (n = 42 279) and 8 RCTs for hip fractures (n = 40 886) comparing oral vitamin D, with or without calcium, with calcium or placebo. To incorporate adherence to treatment, we multiplied the dose by the percentage of adherence to estimate the mean received dose (dose x adherence) for each trial. RESULTS: The pooled relative risk (RR) was 0.86 (95% confidence interval [CI], 0.77-0.96) for prevention of nonvertebral fractures and 0.91 (95% CI, 0.78-1.05) for the prevention of hip fractures, but with significant heterogeneity for both end points. Including all trials, antifracture efficacy increased significantly with a higher dose and higher achieved blood 25 hydroxyvitamin D levels for both end points. Consistently, pooling trials with a higher received dose of more than 400 IU/d resolved heterogeneity. For the higher dose, the pooled RR was 0.80 (95% CI, 0.72-0.89; n = 33 265 subjects from 9 trials) for nonvertebral fractures and 0.82 (95% CI, 0.69-0.97; n = 31 872 subjects from 5 trials) for hip fractures. The higher dose reduced nonvertebral fractures in community-dwelling individuals (-29%) and institutionalized older individuals (-15%), and its effect was independent of additional calcium supplementation. CONCLUSION: Nonvertebral fracture prevention with vitamin D is dose dependent, and a higher dose should reduce fractures by at least 20% for individuals aged 65 years or older. PMID- 19307518 TI - Meat intake and mortality: a prospective study of over half a million people. AB - BACKGROUND: High intakes of red or processed meat may increase the risk of mortality. Our objective was to determine the relations of red, white, and processed meat intakes to risk for total and cause-specific mortality. METHODS: The study population included the National Institutes of Health-AARP (formerly known as the American Association of Retired Persons) Diet and Health Study cohort of half a million people aged 50 to 71 years at baseline. Meat intake was estimated from a food frequency questionnaire administered at baseline. Cox proportional hazards regression models estimated hazard ratios (HRs) and 95% confidence intervals (CIs) within quintiles of meat intake. The covariates included in the models were age, education, marital status, family history of cancer (yes/no) (cancer mortality only), race, body mass index, 31-level smoking history, physical activity, energy intake, alcohol intake, vitamin supplement use, fruit consumption, vegetable consumption, and menopausal hormone therapy among women. Main outcome measures included total mortality and deaths due to cancer, cardiovascular disease, injuries and sudden deaths, and all other causes. RESULTS: There were 47 976 male deaths and 23 276 female deaths during 10 years of follow-up. Men and women in the highest vs lowest quintile of red (HR, 1.31 [95% CI, 1.27-1.35], and HR, 1.36 [95% CI, 1.30-1.43], respectively) and processed meat (HR, 1.16 [95% CI, 1.12-1.20], and HR, 1.25 [95% CI, 1.20-1.31], respectively) intakes had elevated risks for overall mortality. Regarding cause specific mortality, men and women had elevated risks for cancer mortality for red (HR, 1.22 [95% CI, 1.16-1.29], and HR, 1.20 [95% CI, 1.12-1.30], respectively) and processed meat (HR, 1.12 [95% CI, 1.06-1.19], and HR, 1.11 [95% CI 1.04 1.19], respectively) intakes. Furthermore, cardiovascular disease risk was elevated for men and women in the highest quintile of red (HR, 1.27 [95% CI, 1.20 1.35], and HR, 1.50 [95% CI, 1.37-1.65], respectively) and processed meat (HR, 1.09 [95% CI, 1.03-1.15], and HR, 1.38 [95% CI, 1.26-1.51], respectively) intakes. When comparing the highest with the lowest quintile of white meat intake, there was an inverse association for total mortality and cancer mortality, as well as all other deaths for both men and women. CONCLUSION: Red and processed meat intakes were associated with modest increases in total mortality, cancer mortality, and cardiovascular disease mortality. PMID- 19307519 TI - Hypertriglyceridemia and its pharmacologic treatment among US adults. AB - BACKGROUND: Increasing evidence supports triglyceride (TG) concentration as a risk factor for cardiovascular disease. The prevalence of hypertriglyceridemia during a period of rising prevalence of obesity and its pharmacological treatment among US adults are poorly understood. METHODS: We examined data for 5610 participants 20 years or older from the National Health and Nutrition Examination Surveys from 1999 to 2004. RESULTS: The unadjusted prevalence (percentage [SE]) of a TG concentration of 150 mg/dL or higher (to convert triglycerides to millimoles per liter, multiply by 0.0113) was 33.1% (0.8%); a TG concentration of 200 mg/dL or higher, 17.9% (0.7%), a TG concentration of 500 mg/dL or higher, 1.7% (0.2%), and a TG concentration of 1000 mg/dL or higher, 0.4% (0.1%). Overall, 1.3% (0.2%) of participants used 1 of 3 prescription medications indicated to treat hypertriglyceridemia (ie, fenofibrate, gemfibrozil, or niacin); this percentage was 2.6% (0.4%) among participants with a TG concentration of 150 mg/dL or higher and 3.6% (0.7%) among participants with a TG concentration of 200 mg/dL or higher. CONCLUSIONS: Among US adults, hypertriglyceridemia is common. Until the benefits of treating hypertriglyceridemia that is not characterized by extreme elevations of TG concentration with medications are incontrovertible, therapeutic lifestyle change remains the preferred treatment. PMID- 19307520 TI - Hypertriglyceridemia and its pharmacologic treatment among US adults--invited commentary. PMID- 19307521 TI - Body mass index and hypertension hemodynamic subtypes in the adult US population. AB - BACKGROUND: Obesity produces various hemodynamic abnormalities that may impact hypertension subtypes. Similarly, the study of hypertension subtypes provides important information regarding the relative importance of hemodynamic abnormalities contributing to obesity-related hypertension. METHODS: Cross sectional analysis of adults enrolled in the Third National Health and Nutrition Examination Survey (NHANES III) (n = 16 545) and NHANES 1999-2004 (n = 12 137). We examined the relationship between body mass index (BMI) (calculated as weight in kilograms divided by height in meters squared) and the risk of hypertension and hemodynamic subtypes: isolated systolic hypertension (ISH), isolated diastolic hypertension (IDH), and systodiastolic hypertension (SDH). RESULTS: In NHANES 1999-2004, the odds ratio (OR) for hypertension for every 5-unit increase in BMI was 1.45 (95% confidence interval [CI], 1.39-1.52) (P < .001). However, the magnitude of the relative increase in the odds of hypertension was higher among younger adults. Among patients with hypertension, increasing BMI was a significant predictor of IDH or SDH (OR for IDH or SDH, 1.04; 95% CI, 1.02-1.06) (P < .001), as opposed to ISH. Isolated systolic hypertension represented a minority of hypertension cases in obese men (38.9%; 95% CI, 30.9-47.6) but remained the most prevalent type in obese women (62.1%; 95% CI, 52.4%-71.0%) (P < .001), despite a significant relative decrease in the frequency of ISH with increasing BMI in both sexes. Findings in NHANES III were very similar. CONCLUSIONS: Isolated diastolic hypertension and SDH account for most cases of obesity-related hypertension, suggesting that determinants of mean arterial pressure account for the major burden of obesity-related hypertension in US adult men. These findings should be considered in the design of clinical trials and therapeutic strategies for obesity-related hypertension. Further studies should assess determinants of mean arterial pressure in obesity and the role of sex in the pathogenesis of obesity-related hypertension. PMID- 19307522 TI - Red blood cell distribution width and mortality risk in a community-based prospective cohort. AB - BACKGROUND: Red blood cell distribution width (RDW), an automated measure of red blood cell size heterogeneity (eg, anisocytosis) that is largely overlooked, is a newly recognized risk marker in patients with established cardiovascular disease (CVD). It is unknown whether RDW is associated with mortality in the general population or whether this association is specific to CVD. METHODS: We examined the association of RDW with all-cause mortality and with CVD, cancer, and chronic lower respiratory tract disease mortality in 15 852 adult participants in the Third National Health and Nutrition Examination Survey (1988-1994), a nationally representative sample of the US population. Mortality status was obtained by matching to the National Death Index, with follow-up through December 31, 2000. RESULTS: Estimated mortality rates increased 5-fold from the lowest to the highest quintile of RDW after accounting for age and 2-fold after multivariable adjustment (P(trend) < .001 for each). A 1-SD increment in RDW (0.98%) was associated with a 23% greater risk of all-cause mortality (hazard ratio [HR], 1.23; 95% confidence interval [CI], 1.18-1.28) after multivariable adjustment. The RDW was also associated with risk of death due to CVD (HR, 1.22; 95% CI, 1.14 1.31), cancer (1.28; 1.21-1.36), and chronic lower respiratory tract disease (1.32; 1.17-1.49). CONCLUSIONS: Higher RDW is associated with increased mortality risk in this large, community-based sample, an association not specific to CVD. Study of anisocytosis may, therefore, yield novel pathophysiologic insights, and measurement of RDW may contribute to risk assessment. PMID- 19307523 TI - A population-based study of anesthesia consultation before major noncardiac surgery. AB - BACKGROUND: In single-center studies, consultation by an anesthesiologist days to weeks before surgery was associated with reduced patient anxiety, case cancellations on the day of surgery, and duration of hospitalization. Nonetheless, the impact of anesthesia consultation on outcomes in the population remains unclear. METHODS: We used population-based, linked, administrative databases to conduct a cohort study of patients, aged 40 years and older, who underwent selected elective intermediate- to high-risk noncardiac surgical procedures in Ontario, Canada, between April 1, 1994, and March 31, 2004. Propensity-score methods were used to construct a matched-pairs cohort that resolved important differences between patients who underwent consultation and those who did not. We then determined the association of consultation (within 60 days before surgery) with hospital length of stay and postoperative mortality (30 day and 1-year) rates within the matched pairs. RESULTS: Of the 271 082 patients in the entire cohort, 39% (n = 104 716) underwent anesthesia consultation. The proportion of patients who underwent consultation increased from 19% in 1994 to 53% in 2003. Within the matched-pairs (n = 180 254), consultation was associated with reduced mean hospital length of stay (8.17 days vs 8.52 days; difference, 0.35 days; 95% confidence interval [CI], -0.27 to -0.43; P < .001). Consultation was not associated with reduced mortality at 30 days (relative risk, 1.04; 95% CI, 0.96 to 1.13; P = .36) or 1 year (relative risk, 0.98; 95% CI, 0.95 to 1.02; P = .20). CONCLUSIONS: Preoperative anesthesia consultation is associated with reduced length of stay but not with reduced mortality. Future research should evaluate the cost-effectiveness of the increasing use of anesthesia consultation. PMID- 19307524 TI - Smoking and risk of acute and chronic pancreatitis among women and men: a population-based cohort study. AB - BACKGROUND: Alcohol and gallstone disease are the most established risk factors for pancreatitis. Smoking is rarely considered to be a cause despite the fact that a few studies have indicated the opposite. We aimed to assess the independent effects of smoking on the risk of pancreatitis. METHODS: We used data from an observational, population-based cohort study conducted in Denmark. Participants were 9573 women and 8332 men who were followed up for a mean of 20.2 years. Participants underwent a physical examination and completed self administered questionnaires about lifestyle habits. Information on incident cases of acute and chronic pancreatitis were obtained by record linkage with the Danish national registries. RESULTS: A total of 235 cases of pancreatitis occurred during follow-up. A dose-response association between smoking and risk of acute and chronic pancreatitis was observed in both men and women. For example, the hazard ratio of developing pancreatitis was 2.6 (95% confidence interval [CI], 1.5-4.7) among women and 2.6 (95% CI, 1.1-6.2) among men who smoked 15 to 24 grams of tobacco per day. Alcohol intake was associated with an increased risk of pancreatitis (hazard ratio, 1.09; 95% CI, 1.04-1.14 for each additional drink per day). The risk of pancreatitis associated with smoking, however, was independent of alcohol and gallstone disease. Approximately 46% of cases of pancreatitis were attributable to smoking in this cohort. CONCLUSION: In this population of Danish men and women, smoking was independently associated with increased risk of pancreatitis. PMID- 19307525 TI - The value of family history as a risk indicator for venous thrombosis. AB - BACKGROUND: A positive family history of venous thrombosis may reflect the presence of genetic risk factors. Once a risk factor has been identified, it is not known whether family history is of additional value in predicting an individual's risk. We studied the contribution of family history to the risk of venous thrombosis in relation to known risk factors. METHODS: In the Multiple Environmental and Genetic Assessment of risk factors for venous thrombosis, a population-based case-control study, we collected blood samples and information about family history and environmental triggers from 1605 patients with a first venous thrombosis and 2159 control subjects. RESULTS: A total of 505 patients (31.5%) and 373 controls (17.3%) reported having 1 or more first-degree relatives with a history of venous thrombosis. A positive family history increased the risk of venous thrombosis more than 2-fold (odds ratio [95% confidence interval], 2.2 [1.9-2.6]) and up to 4-fold (3.9 [2.7-5.7]) when more than 1 relative was affected. Family history corresponded poorly with known genetic risk factors. Both in those with and without genetic or environmental risk factors, family history remained associated with venous thrombosis. The risk increased with the number of factors identified; for those with a genetic and environmental risk factor and a positive family history, the risk was about 64-fold higher than for those with no known risk factor and a negative family history. CONCLUSIONS: Family history is a risk indicator for a first venous thrombosis, regardless of the other risk factors identified. In clinical practice, family history may be more useful for risk assessment than thrombophilia testing. PMID- 19307526 TI - Long-term effects of metformin on metabolism and microvascular and macrovascular disease in patients with type 2 diabetes mellitus. AB - BACKGROUND: We investigated whether metformin hydrochloride has sustained beneficial metabolic and (cardio) vascular effects in patients with type 2 diabetes mellitus (DM2). METHODS: We studied 390 patients treated with insulin in the outpatient clinics of 3 hospitals in a randomized, placebo-controlled trial with a follow-up period of 4.3 years. Either metformin hydrochloride, 850 mg, or placebo (1-3 times daily) was added to insulin therapy. The primary end point was an aggregate of microvascular and macrovascular morbidity and mortality. The secondary end points were microvascular and macrovascular morbidity and mortality, as separate aggregate scores. In addition, effects on hemoglobin A(1c) (HbA(1c)), insulin requirement, lipid levels, blood pressure, body weight, and body mass index were analyzed. RESULTS: Metformin treatment prevented weight gain (mean weight gain, -3.07 kg [range, -3.85 to -2.28 kg]; P < .001), improved glycemic control (mean reduction in HbA(1c) level, 0.4% percentage point [95% CI, 0.55-0.25]; P < .001) (where CI indicates confidence interval), despite the aim of similar glycemic control in both groups, and reduced insulin requirements (mean reduction, 19.63 IU/d [95% CI, 24.91-14.36 IU/d]; P < .001). Metformin was not associated with an improvement in the primary end point. It was, however, associated with an improvement in the secondary, macrovascular end point (hazard ratio, 0.61 (95% CI, 0.40-0.94; P = .02), which was partly explained by the difference in weight. The number needed to treat to prevent 1 macrovascular end point was 16.1 (95% CI, 9.2-66.6). CONCLUSIONS: Metformin, added to insulin in patients with DM2, improved body weight, glycemic control, and insulin requirements but did not improve the primary end point. Metformin did, however, reduce the risk of macrovascular disease after a follow-up period of 4.3 years. These sustained beneficial effects support the policy to continue metformin treatment after the introduction of insulin in any patient with DM2, unless contraindicated. Trial Registration ClinicalTrials.gov Identifier: NCT00375388. PMID- 19307527 TI - Demographic differences and trends of vitamin D insufficiency in the US population, 1988-2004. AB - BACKGROUND: Vitamin D insufficiency is associated with suboptimal health. The prevalence of vitamin D insufficiency may be rising, but population-based trends are uncertain. We sought to evaluate US population trends in vitamin D insufficiency. METHODS: We compared serum 25-hydroxyvitamin D (25[OH]D) levels from the Third National Health and Nutrition Examination Survey (NHANES III), collected during 1988 through 1994, with NHANES data collected from 2001 through 2004 (NHANES 2001-2004). Complete data were available for 18 883 participants in NHANES III and 13 369 participants in NHANES 2001-2004. RESULTS: The mean serum 25(OH)D level was 30 (95% confidence interval [CI], 29-30) ng/mL during NHANES III and decreased to 24 (23-25) ng/mL during NHANES 2001-2004. Accordingly, the prevalence of 25(OH)D levels of less than 10 ng/mL increased from 2% (95% CI, 2% 2%) to 6% (5%-8%), and 25(OH)D levels of 30 ng/mL or more decreased from 45% (43% 47%) to 23% (20%-26%). The prevalence of 25(OH)D levels of less than 10 ng/mL in non-Hispanic blacks rose from 9% during NHANES III to 29% during NHANES 2001 2004, with a corresponding decrease in the prevalence of levels of 30 ng/mL or more from 12% to 3%. Differences by age strata (mean serum 25[OH]D levels ranging from 28-32 ng/mL) and sex (28 ng/mL for women and 32 ng/mL for men) during NHANES III equalized during NHANES 2001-2004 (24 vs 24 ng/mL for age and 24 vs 24 ng/mL for sex). CONCLUSIONS: National data demonstrate a marked decrease in serum 25(OH)D levels from the 1988-1994 to the 2001-2004 NHANES data collections. Racial/ethnic differences have persisted and may have important implications for known health disparities. Current recommendations for vitamin D supplementation are inadequate to address the growing epidemic of vitamin D insufficiency. PMID- 19307528 TI - Prehospital delays in patients presenting with myocardial infarction. PMID- 19307529 TI - Patients with cancer who are affected by dementia do not die only because of cancer. PMID- 19307530 TI - Support for benefit of physical activity on satiety, weight control, and diabetes risk. PMID- 19307531 TI - Vitamin C concentration and type 2 diabetes mellitus. PMID- 19307532 TI - Progression of the prevalence of metabolic syndrome in normal-weight US subjects. PMID- 19307533 TI - Issues in body fatness measurement. PMID- 19307534 TI - Teaching NeuroImages: cerebral air embolism secondary to atrial-esophageal fistula. PMID- 19307535 TI - Teaching Video NeuroImages: preserved awareness in a frontal seizure with bilateral motor involvement: a psychogenic mimic. PMID- 19307537 TI - Subclinical cortical reorganization: a preclinical imaging marker for recessively inherited PD? PMID- 19307538 TI - CSF biomarkers in relationship to cognitive profiles in Alzheimer disease. AB - OBJECTIVE: To investigate the relationship between CSF biomarkers and cognitive profiles in Alzheimer disease (AD). METHODS: We included 177 patients with AD. Digit Span, Visual Association Test (VAT), VAT object naming, Trail Making Test (TMT), and category fluency were used to assess cognitive functions. Disease severity was assessed using Mini-Mental State Examination; functional impairment was rated by Clinical Dementia Rating. In CSF, levels of amyloid-beta 1-42 (Abeta(1-42)), tau, and tau phosphorylated at threonine 181 (p-tau) were measured. K-means cluster analysis was performed with the three biomarkers to obtain three clusters. Multivariate analysis of variance for repeated measures was performed with CSF cluster as between-subjects factor, neuropsychological z scores as within-subjects variable, and age, sex, and education as covariates. RESULTS: Cluster 1 consisted of 88 patients (49%) with relatively high levels of Abeta(1-42) and low levels of tau and p-tau. Cluster 2 contained 72 patients (41%) with relatively low levels of Abeta(1-42) and high levels of tau and p-tau. Cluster 3 was made up of 17 patients (10%) with low levels of Abeta(1-42) and very high levels of tau and p-tau. No differences between clusters on age, sex, education, APOE genotype, disease duration, functional impairment, or disease severity were found. Patients in cluster 3 performed worse on VAT, TMT-A and -B, and fluency. CONCLUSIONS: Clusters of CSF biomarker levels are related to cognitive profiles in Alzheimer disease. A subgroup of patients with extremely high CSF levels of tau and tau phosphorylated at threonine 181 shows a distinct cognitive profile with more severe impairment of memory, mental speed, and executive functions, which cannot be explained by disease severity. PMID- 19307539 TI - Cerebral hemodynamics and cognitive performance in patients with asymptomatic carotid stenosis. AB - OBJECTIVE: The aim of this study was to investigate whether the presence of severe internal carotid artery stenosis may be associated with different cognitive performance in relation to the side of the stenosis and its hemodynamic consequences. METHODS: Eighty-three patients with asymptomatic severe unilateral internal carotid stenosis were included. A neuropsychological investigation including Verbal Fluency using phonemic and category access, Coloured Progressive Matrices, and Complex Figure Test Copy was performed. Each patient underwent an assessment of cerebrovascular reactivity (CVR) to hypercapnia with transcranial Doppler ultrasonography using the breath-holding index (BHI). Thirty healthy subjects comparable for demographic characteristics and vascular risk profile served as controls. Subjects with carotid stenosis were classified into two groups: preserved CVR (BHI > or =0.69), 48 patients (25 with left and 23 with right stenosis); and impaired CVR (BHI <0.69), 35 patients (19 with left and 16 with right stenosis). RESULTS: Subjects with left stenosis and reduced CVR had significantly lower performances at phonemic verbal fluency with respect to controls and the other groups of stenosis. In subjects with right stenosis and reduced CVR, scores obtained in Coloured Progressive Matrices and in Complex Figure Test Copy were significantly lower with respect to the other groups. CONCLUSIONS: These results suggest that an alteration of cerebrovascular reactivity may be responsible for reduction in some cognitive abilities involving the function of the hemisphere ipsilateral to carotid stenosis. Such findings may be of interest for providing a more comprehensive indication to surgical treatment in subgroups of subjects with asymptomatic carotid stenosis. PMID- 19307540 TI - Osteopenia and osteoporosis in idiopathic benign positional vertigo. AB - OBJECTIVE: Causes of benign positional vertigo (BPV) are mostly unknown. The aim of this study was to elucidate an association of osteoporosis with idiopathic BPV. METHODS: Two hundred nine consecutive patients with a confirmed diagnosis of idiopathic BPV underwent bone mineral densitometry of anterior-posterior lumbar spine and femur. The T scores were compared with those of 202 controls without a history of dizziness. Recurrence was defined when the patients reported two or more previous episodes of positional vertigo similar to those experienced at the time of diagnosis. RESULTS: In both women and men, the lowest T scores were decreased in patients with BPV compared with those in controls. Furthermore, the prevalences of osteopenia (-2.5 < T score < -1.0) and osteoporosis (T score < or =-2.5) were higher in both women and men with BPV than in controls. Multiple logistic regression analyses adjusted for age, sex, alcohol, smoking, and hyperphosphatemia showed that only the existence of osteopenia/osteoporosis was associated with an increased risk of BPV (adjusted odds ratio of osteopenia = 2.0, 95% confidence interval 1.2-3.4, p = 0.011; adjusted odds ratio of osteoporosis = 3.1, 95% confidence interval 1.4-7.2, p = 0.007). In women aged > or =45 years, the lowest T scores were also decreased in the recurrent group, compared with those in the de novo group. CONCLUSION: Osteopenia/osteoporosis may be associated with idiopathic benign positional vertigo (BPV). The effectiveness of measuring bone mineral densitometry and restoring normal calcium metabolism for preventing recurrences of BPV requires further validation. PMID- 19307541 TI - Optical coherence tomography differs in neuromyelitis optica compared with multiple sclerosis. AB - BACKGROUND: Neuromyelitis optica (NMO) is associated with destructive inflammatory lesions, resulting in necrosis and axonal injury. Disability from multiple sclerosis (MS) is due to a combination of demyelination and varying axonal involvement. Optical coherence tomography (OCT), by measuring retinal nerve fiber layer (RNFL) as a surrogate of axonal injury, has potential to discriminate between these two conditions. METHODS: Included were 22 subjects with NMO or NMO spectrum disorders and 47 with MS. Seventeen subjects with NMO and all with MS had a remote history of optic neuritis (ON) in at least one eye, at least 6 months before OCT. Linear mixed modeling was used to compare the two diagnoses for a given level of vision loss, while controlling for age, disease duration, and number of episodes of ON. RESULTS: After ON, NMO was associated with a thinner mean RNFL compared to MS. This was found when controlling for visual acuity (56.7 vs 66.6 microm, p = 0.01) or for contrast sensitivity (61.2 vs 70.3 microm, p = 0.02). The superior and inferior quadrants were more severely affected in NMO than MS. CONCLUSIONS: Optic neuritis (ON) within neuromyelitis optica (NMO) is associated with a thinner overall average retinal nerve fiber layer compared to multiple sclerosis, with particular involvement of the superior and inferior quadrants. This suggests that NMO is associated with more widespread axonal injury in the affected optic nerves. Optical coherence tomography can help distinguish the etiology of these two causes of ON, and may be useful as a surrogate marker of axonal involvement in demyelinating disease. PMID- 19307542 TI - Saccades in adult Niemann-Pick disease type C reflect frontal, brainstem, and biochemical deficits. AB - BACKGROUND: The autosomal recessive disorder Niemannn-Pick type C (NPC) presents in adulthood with psychosis or cognitive deficits associated with supranuclear gaze palsies. While saccadic innervation to the extraocular muscles is generated in the brainstem, the frontal lobes play an integral role in the initiation of volitional saccades and the suppression of unwanted reflexive saccades. No study has examined the frontally driven volitional control of saccadic eye movements in NPC. OBJECTIVE: To examine self-paced and antisaccades as well as reflexive saccades in adult patients with NPC, a disorder known to affect brainstem and frontal cortical function. METHODS: Three biochemically confirmed adult patients with NPC were compared with 10 matched controls on horizontal saccadic and antisaccadic measures using an infrared limbus eye tracker. Patients' cholesterol esterification and filipin staining, Mini-Mental State performance, and NPC symptom level were rated. RESULTS: Reflexive saccade latency ranged from shorter to longer than normal, reflexive saccade gain was reduced, asymptotic peak velocity was reduced, fewer self-paced saccades were generated, and increased errors on antisaccades were made by patients compared to controls. Patients with more severe biochemical, cognitive, and symptom deficits performed most poorly on brainstem and frontal ocular motor measures. Paradoxically, less severe illness was associated with an abnormally reduced saccadic latency. CONCLUSIONS: Ocular motor measures provide an index of disease severity in Niemannn-Pick type C (NPC) and may be a useful adjunct for monitoring the illness progress and medication response. Reduced saccadic latency may result from inadequate fixation input from abnormally functioning frontal eye fields in NPC. PMID- 19307543 TI - Natural history and clinical features of the flail arm and flail leg ALS variants. AB - OBJECTIVE: We sought to define the significance of brachial amyotrophic diplegia (flail arm syndrome [FA]) and the pseudopolyneuritic variant (flail leg syndrome [FL]) of amyotrophic lateral sclerosis (ALS; motor neuron disease). METHODS: We analyzed survival in clinic cohorts in London, UK (1,188 cases), and Melbourne, Australia (432 cases). Survival from disease onset was analyzed using the Kaplan- Meier method and Cox proportional hazards model. RESULTS: In the London cohort, the FA syndrome represented 11% and the FL syndrome 6% of the sample. Median survival was 35 months for limb onset and 27 months for bulbar onset ALS, whereas this was 61 months for FA syndrome (p < 0.001) and 69 months for FL syndrome (p < 0.001). Five-year survival in this cohort was 8.8% for bulbar onset, 20% for limb onset, 52% for FA syndrome, and 64% for FL syndrome. The ratio of men to women was 4:1 in the FA group compared to 2:1 in other limb onset cases. Excluding lower motor neuron FA and FL cases, progressive muscular atrophy comprised 4% of the sample and had a prognosis similar to typical limb onset ALS. In the Melbourne cohort, median survival for limb onset ALS was 31 months, bulbar onset 27 months, FA syndrome 66 months (p < 0.001), and FL syndrome 71 months (p = 0.001). CONCLUSIONS: The flail arm (FA) and flail leg (FL) syndromes had significantly better survival than typical amyotrophic lateral sclerosis (ALS) or progressive muscular atrophy cases that were not classified as FA or FL. Our findings underline the clinical and prognostic importance of the FA and FL variants of ALS. PMID- 19307544 TI - Do the unintended actions of botulinum toxin at distant sites have clinical implications? AB - Over the past 2 decades, botulinum toxin (BT) has enjoyed phenomenal success as a safe and effective therapeutic tool for neurologic and non-neurologic conditions. Even though recent evidence-based conclusions are limited by the availability of data, clinicians' practice confidently recommends BT for many clinical conditions. Besides being effective, BT injected locally has also been considered safe, because no evidence showed that the toxin acts also at distant sites. Recent findings from basic scientific research now challenge this conviction and raise concern that the toxin may have a less localized function than previously thought. Studies in rodents show that the toxin is retrogradely transported and even transcytosed to second-order neurons in the CNS. We therefore need to reappraise whether BT injected into muscles, glands, or cutis might induce previously unconsidered central actions, and whether these actions might have clinical implications. In eliciting clinical benefits, BT's peripheral and central action probably summate. Whether BT acts centrally mainly through retrograde transport, transcytosis, or both remains unclear. Whatever the mechanism, the lack of deleterious central effects implies that while research into action mechanisms continues, physicians can safely use BT for therapy. PMID- 19307545 TI - 4-Aminopyridine toxicity mimics autoimmune-mediated limbic encephalitis. PMID- 19307546 TI - NMO-IgG detected in CSF in seronegative neuromyelitis optica. PMID- 19307547 TI - Fatal congenital myopathy and gastrointestinal pseudo-obstruction due to POLG1 mutations. PMID- 19307548 TI - Reflections: neurology and the humanities. You had the choice, Martha. PMID- 19307549 TI - Intranasal contact point headache: missing the "point" on brain MRI. PMID- 19307550 TI - Increased frequency of isolated cleft palate in infants exposed to lamotrigine during pregnancy. PMID- 19307551 TI - A longitudinal study of drivers with Alzheimer disease. PMID- 19307552 TI - Subthalamic nucleus and its connections: anatomic substrate for the network effects of deep brain stimulation. PMID- 19307553 TI - Adult deafness induces somatosensory conversion of ferret auditory cortex. AB - In response to early or developmental lesions, responsiveness of sensory cortex can be converted from the deprived modality to that of the remaining sensory systems. However, little is known about capacity of the adult cortex for cross modal reorganization. The present study examined the auditory cortices of animals deafened as adults, and observed an extensive somatosensory conversion within as little as 16 days after deafening. These results demonstrate that cortical cross modal reorganization can occur after the period of sensory system maturation. PMID- 19307554 TI - The capillarity of nanometric water menisci confined inside closed-geometry viral cages. AB - We present an investigation of water menisci confined in closed geometries by studying the structural effects of their capillary forces on viruses during the final stage of desiccation. We used individual particles of the bacteriophage phi29 and the minute virus of mice. In both cases the genomic DNA was ejected from the capsid. However, although the structural integrity of the minute virus of mice was essentially preserved, the phi29 capsid underwent a wall-to-wall collapse. We provide evidence that the capillary forces of water confined inside the viruses are mainly responsible for these effects. Moreover, by performing theoretical simulations with a lattice gas model, we found that some structural differences between these 2 viruses may be crucial to explain the different ways in which they are affected by water menisci forces confined at the nanoscale. PMID- 19307555 TI - The medial prefrontal cortex exhibits money illusion. AB - Behavioral economists have proposed that money illusion, which is a deviation from rationality in which individuals engage in nominal evaluation, can explain a wide range of important economic and social phenomena. This proposition stands in sharp contrast to the standard economic assumption of rationality that requires individuals to judge the value of money only on the basis of the bundle of goods that it can buy-its real value-and not on the basis of the actual amount of currency-its nominal value. We used fMRI to investigate whether the brain's reward circuitry exhibits money illusion. Subjects received prizes in 2 different experimental conditions that were identical in real economic terms, but differed in nominal terms. Thus, in the absence of money illusion there should be no differences in activation in reward-related brain areas. In contrast, we found that areas of the ventromedial prefrontal cortex (vmPFC), which have been previously associated with the processing of anticipatory and experienced rewards, and the valuation of goods, exhibited money illusion. We also found that the amount of money illusion exhibited by the vmPFC was correlated with the amount of money illusion exhibited in the evaluation of economic transactions. PMID- 19307556 TI - On the chimeric nature, thermophilic origin, and phylogenetic placement of the Thermotogales. AB - Since publication of the first Thermotogales genome, Thermotoga maritima strain MSB8, single- and multi-gene analyses have disagreed on the phylogenetic position of this order of Bacteria. Here we present the genome sequences of 4 additional members of the Thermotogales (Tt. petrophila, Tt. lettingae, Thermosipho melanesiensis, and Fervidobacterium nodosum) and a comprehensive comparative analysis including the original T. maritima genome. While ribosomal protein genes strongly place Thermotogales as a sister group to Aquificales, the majority of genes with sufficient phylogenetic signal show affinities to Archaea and Firmicutes, especially Clostridia. Indeed, on the basis of the majority of genes in their genomes (including genes that are also found in Aquificales), Thermotogales should be considered members of the Firmicutes. This result highlights the conflict between the taxonomic goal of assigning every species to a unique position in an inclusive Linnaean hierarchy and the evolutionary goal of understanding phylogenesis in the presence of pervasive horizontal gene transfer (HGT) within prokaryotes. Amino acid compositions of reconstructed ancestral sequences from 423 gene families suggest an origin of this gene pool even more thermophilic than extant members of this order, followed by adaptation to lower growth temperatures within the Thermotogales. PMID- 19307557 TI - Social isolation alters neuroinflammatory response to stroke. AB - Social isolation has dramatic long-term physiological and psychological consequences; however, the mechanisms by which social isolation influences disease outcome are largely unknown. The purpose of the present study was to investigate the effects of social isolation on neuronal damage, neuroinflammation, and functional outcome after focal cerebral ischemia. Male mice were socially isolated (housed individually) or pair housed with an ovariectomized female before induction of stroke, via transient intraluminal middle cerebral artery occlusion (MCAO), or SHAM surgery. In these experiments, peri-ischemic social isolation decreases poststroke survival rate and exacerbates infarct size and edema development. The social influence on ischemic damage is accompanied by an altered neuroinflammatory response; specifically, central interleukin-6 (IL-6) signaling is down-regulated, whereas peripheral IL-6 is up regulated, in isolated relative to socially housed mice. In addition, intracerebroventricular injection of an IL-6 neutralizing antibody (10 ng) eliminates social housing differences in measures of ischemic outcome. Taken together, these data suggest that central IL-6 is an important mediator of social influences on stroke outcome. PMID- 19307558 TI - The endometrium responds differently to cloned versus fertilized embryos. AB - Although somatic cell nuclear transfer (SCNT) cloning is more efficient in cattle than in any other species tested so far, there is a high rate of pregnancy failure that has been linked to structural and functional abnormalities of the placenta. We tested the hypothesis that these changes may originate from disturbed embryo-maternal interactions in the peri-implantation period. Therefore, we evaluated the response of the endometrium to SCNT embryos (produced from 7 different fetal fibroblast cell lines) as compared with embryos derived from in vitro fertilization (IVF). SCNT embryos and IVF embryos were cultured under identical conditions to the blastocyst stage (day 7) and were transferred to corresponding recipients, which were slaughtered at day 18 of pregnancy. The mRNA profiles of endometrium samples were obtained using a custom cDNA microarray enriched for transcripts differentially expressed in the endometrium and/or oviduct epithelium during the estrous cycle and/or early pregnancy. Overall, the variation in mRNA profiles was greater in the SCNT group than in the IVF group. Furthermore, 58 transcripts were differentially abundant in endometria from SCNT and IVF pregnancies. Prominent examples are orphan nuclear receptor COUP-TFII and connexin 43, both known to play important roles in uterine receptivity and conceptus placentation. These findings suggest that placental failure in bovine clone pregnancies may originate from abnormal embryo-maternal communication that develops during the peri-implantation period. Endometrium transcriptome profiles may serve as a tool to evaluate SCNT embryos for their ability to establish pregnancy and develop a functional placenta. PMID- 19307559 TI - COUP-TFII acts downstream of Wnt/beta-catenin signal to silence PPARgamma gene expression and repress adipogenesis. AB - Wnt signaling through beta-catenin and TCF maintains preadipocytes in an un differentiated proliferative state; however, the molecular pathway has not been completely defined. By integrating gene expression microarray, chromatin immunoprecipitation-chip, and cell-based experimental approaches, we show that Wnt/beta-catenin signaling activates the expression of COUP-TFII which recruits the SMRT corepressor complex to the first introns located downstream from the first exons of both PPARgamma1 and gamma2 mRNAs. This maintains the local chromatin in a hypoacetylated state and represses PPARgamma gene expression to inhibit adipogenesis. Our experiments define the COUP-TFII/SMRT complex as a previously unappreciated component of the linear pathway that directly links Wnt/beta-catenin signaling to repression of PPARgamma gene expression and the inhibition of adipogenesis. PMID- 19307560 TI - Design of a mechanical clutch-based needle-insertion device. AB - Insertion of trocars, needles, and catheters into unintended tissues or tissue compartments results in hundreds of thousands of complications annually. Current methods for blood vessel cannulation or epidural, chest tube, and initial trocar placement often involve the blind pass of a needle through several layers of tissue and generally rely on distinguishable anatomic landmarks and a high degree of clinical skill. To address this simply and without the use of electronics, a purely mechanical clutch system was developed for use in medical devices that access tissue and tissue compartments. This clutch utilizes the surface contact of a buckled filament inside an S-shaped tube to transmit force from the filament (catheter/guide wire) to the tube (needle). Upon encountering sufficient resistance at the tip, such as dense tissue, the catheter buckles and locks within the tube, causing the filament and needle to advance as one. When the needle reaches the target tissue or fluid-filled cavity, the filament unlocks and slides freely into the target region while the needle remains stationary. A similar locking phenomenon has long been observed in drill strings inside drill shafts used by the oil-drilling industry, and oil industry models were adapted to describe the motion of this clutch system. A predictive analytical model was generated and validated with empirical data and used to develop prototypes of a complete device then tested in vitro on muscle tissue and in vivo on a porcine laparoscopic model with promising results. PMID- 19307561 TI - An intertidal snail shows a dramatic size increase over the past century. AB - Changes in the shell architecture of marine snails enhance defenses and greatly improve survival against predators. In the northwest Atlantic Ocean, shorter and thicker shells have been reported for several species following the introduction of predatory Carcinus maenas crabs early in the 20th century. But we report that the shell lengths of Nucella lapillus actually increased by an average of 22.6% over the past century, with no evidence of shell thickening after correcting for shell length. The increases in shell length were greatest on sheltered shores, highlighting the interaction between wave exposure and the sampling period. Comparisons were based on archived shells collected in 1915-1922 from sites that were resampled in 2007. N. lapillus is an important member of North Atlantic marine ecosystems, and our results suggest that the impacts of historical changes in species' key morphological traits on marine ecosystems remain underappreciated. PMID- 19307562 TI - Kinetic behavior of the major multidrug efflux pump AcrB of Escherichia coli. AB - Multidrug efflux transporters, especially those that belong to the resistance nodulation-division (RND) family, often show very broad substrate specificity and play a major role both in the intrinsic antibiotic resistance and, with increased levels of expression, in the elevated resistance of Gram-negative bacteria. However, it has not been possible to determine the kinetic behavior of these important pumps so far. This is partly because these pumps form a tripartite complex traversing both the cytoplasmic and outer membranes, with an outer membrane channel and a periplasmic adaptor protein, and it is uncertain if the behavior of an isolated component protein reflects that of the protein in this multiprotein complex. Here we use intact cells of Escherichia coli containing the intact multiprotein complex AcrB-AcrA-TolC, and measure the kinetic constants for various cephalosporins, by assessing the periplasmic concentration of the drug from their rate of hydrolysis by periplasmic beta-lactamase and the rate of efflux as the difference between the influx rate and the hydrolysis rate. Nitrocefin efflux showed a K(m) of about 5 microM with little sign of cooperativity. For other compounds (cephalothin, cefamandole, and cephaloridine) that showed lower affinity to the pump, however, kinetics showed strong positive cooperativity, which is consistent with the rotating catalysis model of this trimeric pump. For the very hydrophilic cefazolin there was little sign of efflux. PMID- 19307563 TI - Interference of mismatch and base excision repair during the processing of adjacent U/G mispairs may play a key role in somatic hypermutation. AB - In eukaryotic mismatch repair (MMR), degradation of the error-containing strand initiates at nicks or gaps that can be up to a kilobase away from the mispair. These discontinuities may be the ends of Okazaki fragments or the 3'-termini of the leading strands during replication, whereas the termini of invading strands may fulfill this role during recombination. Here we show that, in extracts of human cells, MMR can initiate also at sites of ongoing base excision repair. Although unlikely under normal circumstances, this situation may arise in vivo during somatic hypermutation (SHM) and class switch recombination of Ig genes, where activation-induced cytidine deaminase (AID) generates multiple U/G mismatches in the variable or switch regions. Uracil should normally be excised by base excision repair (BER), but we show here that MMR proteins activated by a nearby mismatch interfere with uracil processing to generate long single-stranded gaps. We postulate that, in a subset of the repair events, filling-in of the MMR generated gaps might be catalyzed by the error-prone polymerase-eta, rather than by the high-fidelity polymerase-delta. Because polymerase-eta has a propensity to misinsertions opposite adenine residues, the above mechanism would help explain why SHM affects not only C/G, but also A/T base pairs. PMID- 19307564 TI - Extreme longevity in proteinaceous deep-sea corals. AB - Deep-sea corals are found on hard substrates on seamounts and continental margins worldwide at depths of 300 to approximately 3,000 m. Deep-sea coral communities are hotspots of deep ocean biomass and biodiversity, providing critical habitat for fish and invertebrates. Newly applied radiocarbon age dates from the deep water proteinaceous corals Gerardia sp. and Leiopathes sp. show that radial growth rates are as low as 4 to 35 mum year(-1) and that individual colony longevities are on the order of thousands of years. The longest-lived Gerardia sp. and Leiopathes sp. specimens were 2,742 years and 4,265 years, respectively. The management and conservation of deep-sea coral communities is challenged by their commercial harvest for the jewelry trade and damage caused by deep-water fishing practices. In light of their unusual longevity, a better understanding of deep-sea coral ecology and their interrelationships with associated benthic communities is needed to inform coherent international conservation strategies for these important deep-sea habitat-forming species. PMID- 19307565 TI - Identification of intracellular carriers for the endocannabinoid anandamide. AB - The endocannabinoid anandamide (arachidonoyl ethanolamide, AEA) is an uncharged neuromodulatory lipid that, similar to many neurotransmitters, is inactivated through its cellular uptake and subsequent catabolism. AEA is hydrolyzed by fatty acid amide hydrolase (FAAH), an enzyme localized on the endoplasmic reticulum. In contrast to most neuromodulators, the hydrophilic cytosol poses a diffusional barrier for the efficient delivery of AEA to its site of catabolism. Therefore, AEA likely traverses the cytosol with the assistance of an intracellular carrier that increases its solubility and rate of diffusion. To study this process, AEA uptake and hydrolysis were examined in COS-7 cells expressing FAAH restricted to the endoplasmic reticulum, mitochondria, or the Golgi apparatus. AEA hydrolysis was detectable at the earliest measurable time point (3 seconds), suggesting that COS-7 cells, normally devoid of an endocannabinoid system, possess an efficient cytosolic trafficking mechanism for AEA. Three fatty acid binding proteins (FABPs) known to be expressed in brain were examined as possible intracellular AEA carriers. AEA uptake and hydrolysis were significantly potentiated in N18TG2 neuroblastoma cells after overexpression of FABP5 or FABP7, but not FABP3. Similar results were observed in COS-7 cells stably expressing FAAH. Consistent with the roles of FABP as AEA carriers, administration of the competitive FABP ligand oleic acid or the selective non-lipid FABP inhibitor BMS309403 attenuated AEA uptake and hydrolysis by approximately 50% in N18TG2 and COS-7 cells. Taken together, FABPs represent the first proteins known to transport AEA from the plasma membrane to FAAH for inactivation and may therefore be novel pharmacological targets. PMID- 19307566 TI - Pleasant music overcomes the loss of awareness in patients with visual neglect. AB - During the past 20 years there has been much research into the factors that modulate awareness of contralesional information in neurological patients with visual neglect or extinction. However, the potential role of the individual's emotional state in modulating awareness has been largely overlooked. In the current study, we induced a pleasant and positive affective response in patients with chronic visual neglect by allowing them to listen to their pleasant preferred music. We report that the patients showed enhanced visual awareness when tasks were performed under preferred music conditions relative to when tasks were performed either with unpreferred music or in silence. These results were also replicated when positive affect was induced before neglect was tested. Functional MRI data showed enhanced activity in the orbitofrontal cortex and the cingulate gyrus associated with emotional responses when tasks were performed with preferred music relative to unpreferred music. Improved awareness of contralesional (left) targets with preferred music was also associated with a strong functional coupling between emotional areas and attentional brain regions in spared areas of the parietal cortex and early visual areas of the right hemisphere. These findings suggest that positive affect, generated by preferred music, can decrease visual neglect by increasing attentional resources. We discuss the possible roles of arousal and mood in generating these effects. PMID- 19307567 TI - Agricultural origins and the isotopic identity of domestication in northern China. AB - Stable isotope biochemistry (delta(13)C and delta(15)N) and radiocarbon dating of ancient human and animal bone document 2 distinct phases of plant and animal domestication at the Dadiwan site in northwest China. The first was brief and nonintensive: at various times between 7900 and 7200 calendar years before present (calBP) people harvested and stored enough broomcorn millet (Panicum miliaceum) to provision themselves and their hunting dogs (Canis sp.) throughout the year. The second, much more intensive phase was in place by 5900 calBP: during this time both broomcorn and foxtail (Setaria viridis spp. italica) millets were cultivated and made significant contributions to the diets of people, dogs, and pigs (Sus sp.). The systems represented in both phases developed elsewhere: the earlier, low-intensity domestic relationship emerged with hunter-gatherers in the arid north, while the more intensive, later one evolved further east and arrived at Dadiwan with the Yangshao Neolithic. The stable isotope methodology used here is probably the best means of detecting the symbiotic human-plant-animal linkages that develop during the very earliest phases of domestication and is thus applicable to the areas where these connections first emerged and are critical to explaining how and why agriculture began in East Asia. PMID- 19307568 TI - Early modern human diversity suggests subdivided population structure and a complex out-of-Africa scenario. AB - The interpretation of genetic evidence regarding modern human origins depends, among other things, on assessments of the structure and the variation of ancient populations. Because we lack genetic data from the time when the first anatomically modern humans appeared, between 200,000 and 60,000 years ago, instead we exploit the phenotype of neurocranial geometry to compare the variation in early modern human fossils with that in other groups of fossil Homo and recent modern humans. Variation is assessed as the mean-squared Procrustes distance from the group average shape in a representation based on several hundred neurocranial landmarks and semilandmarks. We find that the early modern group has more shape variation than any other group in our sample, which covers 1.8 million years, and that they are morphologically similar to recent modern humans of diverse geographically dispersed populations but not to archaic groups. Of the currently competing models of modern human origins, some are inconsistent with these findings. Rather than a single out-of-Africa dispersal scenario, we suggest that early modern humans were already divided into different populations in Pleistocene Africa, after which there followed a complex migration pattern. Our conclusions bear implications for the inference of ancient human demography from genetic models and emphasize the importance of focusing research on those early modern humans, in particular, in Africa. PMID- 19307569 TI - Auditory recognition memory is inferior to visual recognition memory. AB - Visual memory for scenes is surprisingly robust. We wished to examine whether an analogous ability exists in the auditory domain. Participants listened to a variety of sound clips and were tested on their ability to distinguish old from new clips. Stimuli ranged from complex auditory scenes (e.g., talking in a pool hall) to isolated auditory objects (e.g., a dog barking) to music. In some conditions, additional information was provided to help participants with encoding. In every situation, however, auditory memory proved to be systematically inferior to visual memory. This suggests that there exists either a fundamental difference between auditory and visual stimuli, or, more plausibly, an asymmetry between auditory and visual processing. PMID- 19307570 TI - Starch grain and phytolith evidence for early ninth millennium B.P. maize from the Central Balsas River Valley, Mexico. AB - Questions that still surround the origin and early dispersals of maize (Zea mays L.) result in large part from the absence of information on its early history from the Balsas River Valley of tropical southwestern Mexico, where its wild ancestor is native. We report starch grain and phytolith data from the Xihuatoxtla shelter, located in the Central Balsas Valley, that indicate that maize was present by 8,700 calendrical years ago (cal. B.P.). Phytolith data also indicate an early preceramic presence of a domesticated species of squash, possibly Cucurbita argyrosperma. The starch and phytolith data also allow an evaluation of current hypotheses about how early maize was used, and provide evidence as to the tempo and timing of human selection pressure on 2 major domestication genes in Zea and Cucurbita. Our data confirm an early Holocene chronology for maize domestication that has been previously indicated by archaeological and paleoecological phytolith, starch grain, and pollen data from south of Mexico, and reshift the focus back to an origin in the seasonal tropical forest rather than in the semiarid highlands. PMID- 19307571 TI - The role of symmetry in the mass independent isotope effect in ozone. AB - Understanding the internal distribution of "anomalous" isotope enrichments has important implications for validating theoretical postulates on the origin of these enrichments in molecules such as ozone and for understanding the transfer of these enrichments to other compounds in the atmosphere via mass transfer. Here, we present an approach, using the reaction NO(2)(-) + O(3), for assessing the internal distribution of the Delta(17)O anomaly and the delta(18)O enrichment in ozone produced by electric discharge. The Delta(17)O results strongly support the symmetry mechanism for generating mass independent fractionations, and the delta(18)O results are consistent with published data. Positional Delta(17)O and delta(18)O enrichments in ozone can now be more effectively used in photochemical models that use mass balance oxygen atom transfer mechanisms to infer atmospheric oxidation chemistry. PMID- 19307572 TI - B2 RNA and Alu RNA repress transcription by disrupting contacts between RNA polymerase II and promoter DNA within assembled complexes. AB - Noncoding RNAs (ncRNAs) are now recognized as transregulators of eukaryotic transcription, a role once attributed exclusively to protein factors. Two ncRNAs in mammalian cells have been shown to repress general mRNA transcription by RNA polymerase II (Pol II) in response to heat shock: mouse B2 RNA and human Alu RNA. B2 and Alu RNAs bind directly and tightly to Pol II and co-occupy the promoters of repressed genes along with the polymerase. Here, we identified the molecular mechanism by which mouse B2 RNA and human Alu RNA repress Pol II transcription. Biochemical assays to probe the network of protein-DNA interactions at the promoter revealed that B2 and Alu RNAs prevent Pol II from establishing contacts with the promoter both upstream and downstream of the TATA box during closed complex formation. Disruption of these contacts correlates with transcriptional repression. We conclude that B2 and Alu RNA prevent Pol II from properly engaging the DNA during closed complex formation, resulting in complexes with an altered conformation that are transcriptionally inert. In the absence of its normal contacts with the promoter, Pol II is likely held in these inactive complexes on DNA through interactions with promoter-bound TATA box-binding protein and transcription factor IIB. PMID- 19307573 TI - The cultural and chronological context of early Holocene maize and squash domestication in the Central Balsas River Valley, Mexico. AB - Molecular evidence indicates that the wild ancestor of maize is presently native to the seasonally dry tropical forest of the Central Balsas watershed in southwestern Mexico. We report here on archaeological investigations in a region of the Central Balsas located near the Iguala Valley in Guerrero state that show for the first time a long sequence of human occupation and plant exploitation reaching back to the early Holocene. One of the sites excavated, the Xihuatoxtla Shelter, contains well-stratified deposits and a stone tool assemblage of bifacially flaked points, simple flake tools, and numerous handstones and milling stone bases radiocarbon dated to at least 8700 calendrical years B.P. As reported in a companion paper (Piperno DR, et al., in this issue of PNAS), starch grain and phytolith residues from the ground and chipped stone tools, plus phytoliths from directly associated sediments, provide evidence for maize (Zea mays L.) and domesticated squash (Cucurbita spp.) in contexts contemporaneous with and stratigraphically below the 8700 calendrical years B.P. date. The radiocarbon determinations, stratigraphic integrity of Xihuatoxtla's deposits, and characteristics of the stone tool assemblages associated with the maize and squash remains all indicate that these plants were early Holocene domesticates. Early agriculture in this region of Mexico appears to have involved small groups of cultivators who were shifting their settlements seasonally and engaging in a variety of subsistence pursuits. PMID- 19307574 TI - The mismatch repair system promotes DNA polymerase zeta-dependent translesion synthesis in yeast. AB - DNA lesions that block replication can be bypassed by error-prone or error-free mechanisms. Error-prone mechanisms rely on specialized translesion synthesis (TLS) DNA polymerases that directly replicate over the lesion, whereas error-free pathways use an undamaged duplex as a template for lesion bypass. In the yeast Saccharomyces cerevisiae, most mutagenic TLS of spontaneous and induced DNA damage relies on DNA polymerase zeta (Polzeta) activity. Here, we use a distinct mutational signature produced by Polzeta in a frameshift-reversion assay to examine the role of the yeast mismatch repair (MMR) system in regulating Polzeta dependent mutagenesis. Whereas MMR normally reduces mutagenesis by removing errors introduced by replicative DNA polymerases, we find that the MMR system is required for Polzeta-dependent mutagenesis. In the absence of homologous recombination, however, the error-prone Polzeta pathway is not affected by MMR status. These results demonstrate that MMR promotes Polzeta-dependent mutagenesis by inhibiting an alternative, error-free pathway that depends on homologous recombination. Finally, in contrast to its ability to remove mistakes made by replicative DNA polymerases, we show that MMR fails to efficiently correct errors introduced by Polzeta. PMID- 19307575 TI - A membrane interferometer. AB - Freestanding phospholipid bilayers have been assembled spanning shallow, micrometer-sized wells etched into a Si wafer substrate so that the bilayers are near (within hundreds of nanometers) but not in contact with the wafer surface. The proximity of the bilayers to the highly reflective Si/SiO(2) interface allows them to be probed by using fluorescence-interference techniques. These interferometry measurements show that the bilayers are curved and that the curvature can be varied by changes in osmotic pressure. Furthermore, the ionophore gramicidin can be incorporated into the bilayers, making them selectively permeable to monovalent cations. This freestanding architecture may overcome surface-interaction problems that occur when cell membrane proteins are introduced into solid supported bilayers, while also allowing for high-precision measurements of changes in fluorophore position by interferometry. PMID- 19307576 TI - Posttranscriptional regulation of interleukin-10 expression by hsa-miR-106a. AB - IL-10 is a key regulator of the immune system that critically determines health and disease. Its expression is finely tuned both at the transcriptional and posttranscriptional levels. Although the importance of posttranscriptional regulation of IL-10 has been previously shown, understanding the underlying mechanisms is still in its infancy. In this study, using a combination of bioinformatics and molecular approaches, we report that microRNA (hsa-miR-106a) regulates IL-10 expression. The hsa-miR-106a binding site in the 3' UTR of IL10 has been identified by site-directed mutagenesis studies. Also, the involvement of transcription factors, Sp1 and Egr1, in the regulation of hsa-miR-106a expression and concomitant decrease in the IL-10 expression, has also been demonstrated. In summary, our results showed that IL-10 expression may be regulated by miR-106a, which is in turn transcriptionally regulated by Egr1 and Sp1. PMID- 19307577 TI - Longitudinal trajectories of non-rapid eye movement delta and theta EEG as indicators of adolescent brain maturation. AB - It is now recognized that extensive maturational changes take place in the human brain during adolescence, and that the trajectories of these changes are best studied longitudinally. We report the first longitudinal study of the adolescent decline in non-rapid eye movement (NREM) delta (1-4 Hz) and theta (4-8 Hz) EEG. Delta and theta are the homeostatic frequencies of human sleep. We recorded sleep EEG in 9- and 12-year-old cohorts twice yearly over a 5-year period. Delta power density (PD) was unchanged between age 9 and 11 years and then fell precipitously, decreasing by 66% between age 11 and 16.5 years (P < .000001). The decline in theta PD began significantly earlier than that in delta PD and also was very steep (by 60%) between age 11 and 16.5 years (P < .000001). These data suggest that age 11-16.5 years is a critically important maturational period for the brain processes that underlie homeostatic NREM EEG, a finding not suggested in previous cross-sectional data. We hypothesize that these EEG changes reflect synaptic pruning. Comparing our data with published longitudinal declines in MRI estimated cortical thickness suggests the theta age curve parallels the earlier maturational thinning in 3-layer cortex, whereas the delta curve tracks the later changes in 5-layer cortex. This comparison also reveals that adolescent declines in NREM delta and theta are substantially larger than decreases in cortical thickness (>60% vs. <20%). The magnitude, interindividual difference, and tight link to age of these EEG changes indicate that they provide excellent noninvasive tools for investigating neurobehavioral correlates of adolescent brain maturation. PMID- 19307578 TI - Mammalian cell penetration, siRNA transfection, and DNA transfection by supercharged proteins. AB - Nucleic acid reagents, including small interfering RNA (siRNA) and plasmid DNA, are important tools for the study of mammalian cells and are promising starting points for the development of new therapeutic agents. Realizing their full potential, however, requires nucleic acid delivery reagents that are simple to prepare, effective across many mammalian cell lines, and nontoxic. We recently described the extensive surface mutagenesis of proteins in a manner that dramatically increases their net charge. Here, we report that superpositively charged green fluorescent proteins, including a variant with a theoretical net charge of +36 (+36 GFP), can penetrate a variety of mammalian cell lines. Internalization of +36 GFP depends on nonspecific electrostatic interactions with sulfated proteoglycans present on the surface of most mammalian cells. When +36 GFP is mixed with siRNA, protein-siRNA complexes approximately 1.7 mum in diameter are formed. Addition of these complexes to five mammalian cell lines, including four that are resistant to cationic lipid-mediated siRNA transfection, results in potent siRNA delivery. In four of these five cell lines, siRNA transfected by +36 GFP suppresses target gene expression. We show that +36 GFP is resistant to proteolysis, is stable in the presence of serum, and extends the serum half-life of siRNA and plasmid DNA with which it is complexed. A variant of +36 GFP can mediate DNA transfection, enabling plasmid-based gene expression. These findings indicate that superpositively charged proteins can overcome some of the key limitations of currently used transfection agents. PMID- 19307579 TI - Glucocorticoid regulation of preproglucagon transcription and RNA stability during stress. AB - Stress elicits a synchronized response of the endocrine, sympathetic, and central nervous systems to preserve homeostasis and well-being. Glucagon-like peptide-1 (GLP-1), a primary posttranslational product of the preproglucagon (PPG) gene, activates both physical and psychological stress responses. The current study examined mechanisms regulating expression of PPG gene products in the hindbrain. Our results indicate that PPG mRNA decreases rapidly after exposure to acute stressors of multiple modalities. Reduced mRNA levels are accompanied by reduced GLP-1 immunoreactivity in the paraventricular nucleus of hypothalamus, suggesting release at PPG terminals. Stress-induced decrements in PPG mRNA were attenuated in adrenalectomized-corticosterone-replaced rats, suggesting that mRNA down regulation is due at least in part to glucocorticoid secretion. In contrast, acute stress increased levels of PPG heteronuclear RNA (hnRNA) in a glucocorticoid-dependent manner, suggesting that decreases in PPG mRNA are due to increased degradation rather than reduced transcription. Glucocorticoid administration to unstressed rats is sufficient to cause decrements in PPG mRNA and increments in PPG hnRNA. These findings suggest that glucocorticoids deplete the pool of transcribed PPG mRNA and concurrently stimulate PPG gene transcription, with the latter allowing a mechanism for replenishment of PPG mRNA after stress cessation. The combination of rapid PPG mRNA depletion and initiation of PPG transcription within 30 min is consistent with a rapid action of glucocorticoids on GLP-1 bioavailability, resulting in a transient reduction in the capacity for neuropeptidergic excitation of stress responses. PMID- 19307581 TI - The mosaic genome structure of the Wolbachia wRi strain infecting Drosophila simulans. AB - The obligate intracellular bacterium Wolbachia pipientis infects around 20% of all insect species. It is maternally inherited and induces reproductive alterations of insect populations by male killing, feminization, parthenogenesis, or cytoplasmic incompatibility. Here, we present the 1,445,873-bp genome of W. pipientis strain wRi that induces very strong cytoplasmic incompatibility in its natural host Drosophila simulans. A comparison with the previously sequenced genome of W. pipientis strain wMel from Drosophila melanogaster identified 35 breakpoints associated with mobile elements and repeated sequences that are stable in Drosophila lines transinfected with wRi. Additionally, 450 genes with orthologs in wRi and wMel were sequenced from the W. pipientis strain wUni, responsible for the induction of parthenogenesis in the parasitoid wasp Muscidifurax uniraptor. The comparison of these A-group Wolbachia strains uncovered the most highly recombining intracellular bacterial genomes known to date. This was manifested in a 500-fold variation in sequence divergences at synonymous sites, with different genes and gene segments supporting different strain relationships. The substitution-frequency profile resembled that of Neisseria meningitidis, which is characterized by rampant intraspecies recombination, rather than that of Rickettsia, where genes mostly diverge by nucleotide substitutions. The data further revealed diversification of ankyrin repeat genes by short tandem duplications and provided examples of horizontal gene transfer across A- and B-group strains that infect D. simulans. These results suggest that the transmission dynamics of Wolbachia and the opportunity for coinfections have created a freely recombining intracellular bacterial community with mosaic genomes. PMID- 19307580 TI - Recent speciation of Capsella rubella from Capsella grandiflora, associated with loss of self-incompatibility and an extreme bottleneck. AB - Flowering plants often prevent selfing through mechanisms of self-incompatibility (S.I.). The loss of S.I. has occurred many times independently, because it provides short-term advantages in situations where pollinators or mates are rare. The genus Capsella, which is closely related to Arabidopsis, contains a pair of closely related diploid species, the self-incompatible Capsella grandiflora and the self-compatible Capsella rubella. To elucidate the transition to selfing and its relationship to speciation of C. rubella, we have made use of comparative sequence information. Our analyses indicate that C. rubella separated from C. grandiflora recently ( approximately 30,000-50,000 years ago) and that breakdown of S.I. occurred at approximately the same time. Contrasting the nucleotide diversity patterns of the 2 species, we found that C. rubella has only 1 or 2 alleles at most loci, suggesting that it originated through an extreme population bottleneck. Our data are consistent with diploid speciation by a single, selfing individual, most likely living in Greece. The new species subsequently colonized the Mediterranean by Northern and Southern routes, at a time that also saw the spread of agriculture. The presence of phenotypic diversity within modern C. rubella suggests that this species will be an interesting model to understand divergence and adaptation, starting from very limited standing genetic variation. PMID- 19307582 TI - A family of thermostable fungal cellulases created by structure-guided recombination. AB - SCHEMA structure-guided recombination of 3 fungal class II cellobiohydrolases (CBH II cellulases) has yielded a collection of highly thermostable CBH II chimeras. Twenty-three of 48 genes sampled from the 6,561 possible chimeric sequences were secreted by the Saccharomyces cerevisiae heterologous host in catalytically active form. Five of these chimeras have half-lives of thermal inactivation at 63 degrees C that are greater than the most stable parent, CBH II enzyme from the thermophilic fungus Humicola insolens, which suggests that this chimera collection contains hundreds of highly stable cellulases. Twenty-five new sequences were designed based on mathematical modeling of the thermostabilities for the first set of chimeras. Ten of these sequences were expressed in active form; all 10 retained more activity than H. insolens CBH II after incubation at 63 degrees C. The total of 15 validated thermostable CBH II enzymes have high sequence diversity, differing from their closest natural homologs at up to 63 amino acid positions. Selected purified thermostable chimeras hydrolyzed phosphoric acid swollen cellulose at temperatures 7 to 15 degrees C higher than the parent enzymes. These chimeras also hydrolyzed as much or more cellulose than the parent CBH II enzymes in long-time cellulose hydrolysis assays and had pH/activity profiles as broad, or broader than, the parent enzymes. Generating this group of diverse, thermostable fungal CBH II chimeras is the first step in building an inventory of stable cellulases from which optimized enzyme mixtures for biomass conversion can be formulated. PMID- 19307583 TI - Nanotechnology approach for drug addiction therapy: gene silencing using delivery of gold nanorod-siRNA nanoplex in dopaminergic neurons. AB - Drug abuse is a worldwide health concern in which addiction involves activation of the dopaminergic signaling pathway in the brain. Here, we introduce a nanotechnology approach that utilizes gold nanorod-DARPP-32 siRNA complexes (nanoplexes) that target this dopaminergic signaling pathway in the brain. The shift in the localized longitudinal plasmon resonance peak of gold nanorods (GNRs) was used to show their interaction with siRNA. Plasmonic enhanced dark field imaging was used to visualize the uptake of these nanoplexes in dopaminergic neurons in vitro. Gene silencing of the nanoplexes in these cells was evidenced by the reduction in the expression of key proteins (DARPP-32, ERK, and PP-1) belonging to this pathway, with no observed cytotoxicity. Moreover, these nanoplexes were shown to transmigrate across an in vitro model of the blood brain barrier (BBB). Therefore, these nanoplexes appear to be suited for brain specific delivery of appropriate siRNA for therapy of drug addiction and other brain diseases. PMID- 19307584 TI - Model of mouth-to-mouth transfer of bacterial lipoproteins through inner membrane LolC, periplasmic LolA, and outer membrane LolB. AB - Outer membrane-specific lipoproteins in Escherichia coli are released from the inner membrane by an ATP-binding cassette transporter, the LolCDE complex, which causes the formation of a soluble complex with a periplasmic molecular chaperone, LolA. LolA then transports lipoproteins to the outer membrane where an outer membrane receptor, LolB, incorporates lipoproteins into the outer membrane. The molecular mechanisms underlying the Lol-dependent lipoprotein sorting have been clarified in detail. However, it remained unclear how Lol factors interact with each other to conduct very efficient lipoprotein transfer in the periplasm where ATP is not available. To address this issue, a photo-reactive phenylalanine analogue, p-benzoyl-phenylalanine, was introduced at various positions of LolA and LolB, of which the overall structures are very similar and comprise an incomplete beta-barrel with a hydrophobic cavity inside. Cells expressing LolA or LolB derivatives containing the above analogue were irradiated with UV for in vivo photo-cross-linking. These analyses revealed a hot area in the same region of LolA and LolB, through which LolA and LolB interact with each other. This area is located at the entrance of the hydrophobic cavity. Moreover, this area in LolA is involved in the interaction with a membrane subunit, LolC, whereas no cross linking occurs between LolA and the other membrane subunit, LolE, or ATP-binding subunit LolD, despite the structural similarity between LolC and LolE. The hydrophobic cavities of LolA and LolB were both found to bind lipoproteins inside. These results indicate that the transfer of lipoproteins through Lol proteins occurs in a mouth-to-mouth manner. PMID- 19307585 TI - Theoretical predictions strongly support decision accuracy as a major driver of ecological specialization. AB - We examine the proposal that the high levels of ecological specialization seen in many animals has been driven by benefits in decision accuracy that accrue from this resource-use strategy. Using artificial analogs of real neural processing (artificial neural networks), we examine the relationship between decision accuracy, level of ecological specialization/generalization, and the punishment/reward for selecting non-host resources. We demonstrate that specialists make more accurate resource-use decisions than generalists when the consequences of using a non-host are neutral or positive but not very positive. Pronounced unsuitability of non-host resources in fact promotes higher decision accuracy in generalists. These unusual predictions can be explained by the special properties of neural processing systems and are entirely consistent with patterns of performance of many specialists in nature, where non-used resources are, curiously, often quite suitable for growth and reproduction. They potentially reconcile the long-observed discrepancy between the presence of high levels of ecological specialization in many animal groups and the absence of strong negative fitness correlations across resources. The strong theoretical support obtained here, and the equally good support in experimental studies elsewhere, should bring the "neural limitations" hypothesis to the forefront of research on the evolutionary determinants of ecological range. PMID- 19307586 TI - Single-round selection yields a unique retroviral envelope utilizing GPR172A as its host receptor. AB - The recognition by a viral envelope of its cognate host-cell receptor is the initial critical step in defining the viral host-range and tissue specificity. This study combines a single-round of selection of a random envelope library with a parallel cDNA screen for receptor function to identify a distinct retroviral envelope/receptor pair. The 11-aa targeting domain of the modified feline leukemia virus envelope consists of a constrained peptide. Critical to the binding of the constrained peptide envelope to its cellular receptor are a pair of internal cysteines and an essential Trp required for maintenance of titers >10(5) lacZ staining units per milliliter. The receptor used for viral entry is the human GPR172A protein, a G-protein-coupled receptor isolated from osteosarcoma cells. The ability to generate unique envelopes capable of using tissue- or disease-specific receptors marks an advance in the development of efficient gene-therapy vectors. PMID- 19307587 TI - Rational development of high-affinity T-cell receptor-like antibodies. AB - T-cell interaction with a target cell is a key event in the adaptive immune response and primarily driven by T-cell receptor (TCR) recognition of peptide-MHC (pMHC) complexes. TCR avidity for a given pMHC is determined by number of MHC molecules, availability of coreceptors, and TCR affinity for MHC or peptide, respectively, with peptide recognition being the most important factor to confer target specificity. Here we present high-resolution crystal structures of 2 Fab antibodies in complex with the immunodominant NY-ESO-1(157-165) peptide analogue (SLLMWITQV) presented by HLA-A*0201 and compare them with a TCR recognizing the same pMHC. Binding to the central methionine-tryptophan peptide motif and orientation of binding were almost identical for Fabs and TCR. As the MW "peg" dominates the contacts between Fab and peptide, we estimated the contributions of individual amino acids between the Fab and peptide to provide the rational basis for a peptide-focused second-generation, high-affinity antibody library. The final Fab candidate achieved better peptide binding by 2 light-chain mutations, giving a 20-fold affinity improvement to 2-4 nM, exceeding the affinity of the TCR by 1,000-fold. The high-affinity Fab when grafted as recombinant TCR on T cells conferred specific killing of HLA-A*0201/NY-ESO-1(157-165) target cells. In summary, we prove that affinity maturation of antibodies mimicking a TCR is possible and provide a strategy for engineering high-affinity antibodies that can be used in targeting specific pMHC complexes for diagnostic and therapeutic purposes. PMID- 19307588 TI - Larval dispersal connects fish populations in a network of marine protected areas. AB - Networks of no-take marine protected areas (MPAs) have been widely advocated for the conservation of marine biodiversity. But for MPA networks to be successful in protecting marine populations, individual MPAs must be self-sustaining or adequately connected to other MPAs via dispersal. For marine species with a dispersive larval stage, populations within MPAs require either the return of settlement-stage larvae to their natal reserve or connectivity among reserves at the spatial scales at which MPA networks are implemented. To date, larvae have not been tracked when dispersing from one MPA to another, and the relative magnitude of local retention and connectivity among MPAs remains unknown. Here we use DNA parentage analysis to provide the first direct estimates of connectivity of a marine fish, the orange clownfish (Amphiprion percula), in a proposed network of marine reserves in Kimbe Bay, Papua New Guinea. Approximately 40% of A. percula larvae settling into anemones in an island MPA at 2 different times were derived from parents resident in the reserve. We also located juveniles spawned by Kimbe Island residents that had dispersed as far as 35 km to other proposed MPAs, the longest distance that marine larvae have been directly tracked. These dispersers accounted for up to 10% of the recruitment in the adjacent MPAs. Our findings suggest that MPA networks can function to sustain resident populations both by local replenishment and through larval dispersal from other reserves. More generally, DNA parentage analysis provides a direct method for measuring larval dispersal for other marine organisms. PMID- 19307589 TI - A differentiation pathway for B1 cells in adult bone marrow. AB - The recent description of a Lin(-)AA4.1(+)CD19(+)B220(Lo/-) B1-specified progenitor (B1P) population in adult marrow adds support for the argument that these unique B cells arise from a distinct lineage. However, the origins of B1P were not investigated and their developmental relationships to conventional B2 cells remain unclear. We now report that B1P development is IL-7Ralpha-dependent, and negatively regulated by Bruton tyrosine kinase. Lymphoid characteristics of B1P were further studied with recombination activating gene (RAG)-1/GFP knock-in, RAG-1/Cre reporter, and VEX transgenic mice. Our results reveal that they are heterogeneous with respect to lymphocyte affiliation. RAG-1(+) early lymphoid progenitors and Lin(-)Sca-1(+)cKit(Lo)IL-7Ralpha(+) common lymphoid progenitors from adult marrow efficiently generated CD19(+)CD45R/B220(Lo/-) cells in vitro and in vivo. Moreover, early lymphoid progenitors and common lymphoid progenitors produced significant numbers of peritoneal CD11b(+)CD5(+) B1a and CD11b(+)CD5(-) B1b cells in vivo. Finally, 2-step transplantation experiments established a differentiation pathway between conventional lymphoid progenitors, B1P, and mature B1 lymphocytes. Thus, our findings indicate that at least some B1P can be produced in adult bone marrow from primitive B2 progenitors, and suggest a developmental relationship between the major categories of B lymphocytes. PMID- 19307590 TI - Binding of an octylglucoside detergent molecule in the second substrate (S2) site of LeuT establishes an inhibitor-bound conformation. AB - The first crystal structure of the neurotransmitter/sodium symporter homolog LeuT revealed an occluded binding pocket containing leucine and 2 Na(+); later structures showed tricyclic antidepressants (TCAs) in an extracellular vestibule approximately 11 A above the bound leucine and 2 Na(+). We recently found this region to be a second binding (S2) site and that binding of substrate to this site triggers Na(+)-coupled substrate symport. Here, we show a profound inhibitory effect of n-octyl-beta-d-glucopyranoside (OG), the detergent used for LeuT crystallization, on substrate binding to the S2 site. In parallel, we determined at 2.8 A the structure of LeuT-E290S, a mutant that, like LeuT-WT, binds 2 substrate molecules. This structure was similar to that of WT and clearly revealed an OG molecule in the S2 site. We also observed electron density at the S2 site in LeuT-WT crystals, and this also was accounted for by an OG molecule in that site. Computational analyses, based on the available crystal structures of LeuT, indicated the nature of structural arrangements in the extracellular region of LeuT that differentiate the actions of substrates from inhibitors bound in the S2 site. We conclude that the current LeuT crystal structures, all of which have been solved in OG, represent functionally blocked forms of the transporter, whereas a substrate bound in the S2 site will promote a different state that is essential for Na(+)-coupled symport. PMID- 19307591 TI - SDF-1alpha stimulates JNK3 activity via eNOS-dependent nitrosylation of MKP7 to enhance endothelial migration. AB - The chemokine stromal cell-derived factor-1alpha (SDF-1alpha) is a pivotal player in angiogenesis. It is capable of influencing such cellular processes as tubulogenesis and endothelial cell migration, yet very little is known about the actual signaling events that mediate SDF-1alpha-induced endothelial cell function. In this report, we describe the identification of an intricate SDF 1alpha-induced signaling cascade that involves endothelial nitric oxide synthase (eNOS), JNK3, and MAPK phosphatase 7 (MKP7). We demonstrate that the SDF-1alpha induced activation of JNK3, critical for endothelial cell migration, depends on the prior activation of eNOS. Specifically, activation of eNOS leads to production of NO and subsequent nitrosylation of MKP7, rendering the phosphatase inactive and unable to inhibit the activation of JNK3. These observations reinforce the importance of nitric oxide and S-nitrosylation in angiogenesis and provide a mechanistic pathway for SDF-1alpha-induced endothelial cell migration. In addition, the discovery of this interactive network of pathways provides novel and unexpected therapeutic targets for angiogenesis-dependent diseases. PMID- 19307592 TI - Transcriptional neoteny in the human brain. AB - In development, timing is of the utmost importance, and the timing of developmental processes often changes as organisms evolve. In human evolution, developmental retardation, or neoteny, has been proposed as a possible mechanism that contributed to the rise of many human-specific features, including an increase in brain size and the emergence of human-specific cognitive traits. We analyzed mRNA expression in the prefrontal cortex of humans, chimpanzees, and rhesus macaques to determine whether human-specific neotenic changes are present at the gene expression level. We show that the brain transcriptome is dramatically remodeled during postnatal development and that developmental changes in the human brain are indeed delayed relative to other primates. This delay is not uniform across the human transcriptome but affects a specific subset of genes that play a potential role in neural development. PMID- 19307593 TI - Signals of recent positive selection in a worldwide sample of human populations. AB - Genome-wide scans for recent positive selection in humans have yielded insight into the mechanisms underlying the extensive phenotypic diversity in our species, but have focused on a limited number of populations. Here, we present an analysis of recent selection in a global sample of 53 populations, using genotype data from the Human Genome Diversity-CEPH Panel. We refine the geographic distributions of known selective sweeps, and find extensive overlap between these distributions for populations in the same continental region but limited overlap between populations outside these groupings. We present several examples of previously unrecognized candidate targets of selection, including signals at a number of genes in the NRG-ERBB4 developmental pathway in non-African populations. Analysis of recently identified genes involved in complex diseases suggests that there has been selection on loci involved in susceptibility to type II diabetes. Finally, we search for local adaptation between geographically close populations, and highlight several examples. PMID- 19307594 TI - Time (and PPAR beta/delta) heals all wounds. PMID- 19307595 TI - Marko Kaksonen: tracking the inner secrets of endocytosis. Interview by Caitlin Sedwick. PMID- 19307596 TI - Reversion-induced LIM interaction with Src reveals a novel Src inactivation cycle. AB - Aberrant Src activation plays prominent roles in cancer progression. However, how Src is activated in cancer cells is largely unknown. Genetic Src-activating mutations are rare and, therefore, are insufficient to account for Src activation commonly found in human cancers. In this study, we show that reversion-induced LIM (RIL), which is frequently lost in colon and other cancers as a result of epigenetic silencing, suppresses Src activation. Mechanistically, RIL suppresses Src activation through interacting with Src and PTPL1, allowing PTPL1-dependent dephosphorylation of Src at the activation loop. Importantly, the binding of RIL to Src is drastically reduced upon Src inactivation. Our results reveal a novel Src inactivation cycle in which RIL preferentially recognizes active Src and facilitates PTPL1-mediated inactivation of Src. Inactivation of Src, in turn, promotes dissociation of RIL from Src, allowing the initiation of a new Src inactivation cycle. Epigenetic silencing of RIL breaks this Src inactivation cycle and thereby contributes to aberrant Src activation in human cancers. PMID- 19307597 TI - NF-kappaB and Snail1a coordinate the cell cycle with gastrulation. AB - The cell cycle needs to strictly coordinate with developmental processes to ensure correct generation of the body plan and different tissues. However, the molecular mechanism underlying the coordination remains largely unknown. In this study, we investigate how the cell cycle coordinates gastrulation cell movements in zebrafish. We present a system to modulate the cell cycle in early zebrafish embryos by manipulating the geminin-Cdt1 balance. Alterations of the cell cycle change the apoptotic level during gastrulation, which correlates with the nuclear level of antiapoptotic nuclear factor kappaB (NF-kappaB). NF-kappaB associates with the Snail1a promoter region on the chromatin and directly activates Snail1a, an important factor controlling cell delamination, which is the initial step of mesendodermal cell movements during gastrulation. In effect, the cell cycle coordinates the delamination of mesendodermal cells through the transcription of Snail1a. Our results suggest a molecular mechanism by which NF-kappaB and Snail1a coordinate the cell cycle through gastrulation. PMID- 19307598 TI - Regulation of epithelial-mesenchymal IL-1 signaling by PPARbeta/delta is essential for skin homeostasis and wound healing. AB - Skin morphogenesis, maintenance, and healing after wounding require complex epithelial-mesenchymal interactions. In this study, we show that for skin homeostasis, interleukin-1 (IL-1) produced by keratinocytes activates peroxisome proliferator-activated receptor beta/delta (PPARbeta/delta) expression in underlying fibroblasts, which in turn inhibits the mitotic activity of keratinocytes via inhibition of the IL-1 signaling pathway. In fact, PPARbeta/delta stimulates production of the secreted IL-1 receptor antagonist, which leads to an autocrine decrease in IL-1 signaling pathways and consequently decreases production of secreted mitogenic factors by the fibroblasts. This fibroblast PPARbeta/delta regulation of the IL-1 signaling is required for proper wound healing and can regulate tumor as well as normal human keratinocyte cell proliferation. Together, these findings provide evidence for a novel homeostatic control of keratinocyte proliferation and differentiation mediated via PPARbeta/delta regulation in dermal fibroblasts of IL-1 signaling. Given the ubiquitous expression of PPARbeta/delta, other epithelial-mesenchymal interactions may also be regulated in a similar manner. PMID- 19307599 TI - N-glycans are direct determinants of CFTR folding and stability in secretory and endocytic membrane traffic. AB - N-glycosylation, a common cotranslational modification, is thought to be critical for plasma membrane expression of glycoproteins by enhancing protein folding, trafficking, and stability through targeting them to the ER folding cycles via lectin-like chaperones. In this study, we show that N-glycans, specifically core glycans, enhance the productive folding and conformational stability of a polytopic membrane protein, the cystic fibrosis transmembrane conductance regulator (CFTR), independently of lectin-like chaperones. Defective N glycosylation reduces cell surface expression by impairing both early secretory and endocytic traffic of CFTR. Conformational destabilization of the glycan deficient CFTR induces ubiquitination, leading to rapid elimination from the cell surface. Ubiquitinated CFTR is directed to lysosomal degradation instead of endocytic recycling in early endosomes mediated by ubiquitin-binding endosomal sorting complex required for transport (ESCRT) adaptors Hrs (hepatocyte growth factor-regulated tyrosine kinase substrate) and TSG101. These results suggest that cotranslational N-glycosylation can exert a chaperone-independent profolding change in the energetic of CFTR in vivo as well as outline a paradigm for the peripheral trafficking defect of membrane proteins with impaired glycosylation. PMID- 19307600 TI - A role for ubiquitin ligases and Spartin/SPG20 in lipid droplet turnover. AB - HECT (homologous to the E6AP C terminus) ubiquitin ligases have diverse functions in eukaryotic cells. In screens for proteins that bind to the HECT ubiquitin ligase WWP1, we identified Spartin, which is also known as SPG20. This protein is truncated in a neurological disease, Troyer syndrome. In this study, we show that SPG20 associates with the surface of lipid droplets (LDs) and can regulate their size and number. SPG20 binds to another LD protein, TIP47, and both proteins compete with an additional LD protein, adipophilin/adipocyte differentiation related protein, for occupancy of LDs. The mutant SPG20 present in Troyer syndrome does not possess these activities. Depletion of SPG20 using RNA interference increases the number and size of LDs when cells are fed with oleic acid. Binding of WWP1 to SPG20 and the consequent ubiquitin transfer remove SPG20 from LDs and reduce the levels of coexpressed SPG20. These experiments suggest functions for ubiquitin ligases and SPG20 in the regulation of LD turnover and potential pathological mechanisms in Troyer syndrome. PMID- 19307601 TI - Galpha12/13 regulate epiboly by inhibiting E-cadherin activity and modulating the actin cytoskeleton. AB - Epiboly spreads and thins the blastoderm over the yolk cell during zebrafish gastrulation, and involves coordinated movements of several cell layers. Although recent studies have begun to elucidate the processes that underlie these epibolic movements, the cellular and molecular mechanisms involved remain to be fully defined. Here, we show that gastrulae with altered Galpha(12/13) signaling display delayed epibolic movement of the deep cells, abnormal movement of dorsal forerunner cells, and dissociation of cells from the blastoderm, phenocopying e cadherin mutants. Biochemical and genetic studies indicate that Galpha(12/13) regulate epiboly, in part by associating with the cytoplasmic terminus of E cadherin, and thereby inhibiting E-cadherin activity and cell adhesion. Furthermore, we demonstrate that Galpha(12/13) modulate epibolic movements of the enveloping layer by regulating actin cytoskeleton organization through a RhoGEF/Rho-dependent pathway. These results provide the first in vivo evidence that Galpha(12/13) regulate epiboly through two distinct mechanisms: limiting E cadherin activity and modulating the organization of the actin cytoskeleton. PMID- 19307602 TI - Akt regulates L-type Ca2+ channel activity by modulating Cavalpha1 protein stability. AB - The insulin IGF-1-PI3K-Akt signaling pathway has been suggested to improve cardiac inotropism and increase Ca(2+) handling through the effects of the protein kinase Akt. However, the underlying molecular mechanisms remain largely unknown. In this study, we provide evidence for an unanticipated regulatory function of Akt controlling L-type Ca(2+) channel (LTCC) protein density. The pore-forming channel subunit Ca(v)alpha1 contains highly conserved PEST sequences (signals for rapid protein degradation), and in-frame deletion of these PEST sequences results in increased Ca(v)alpha1 protein levels. Our findings show that Akt-dependent phosphorylation of Ca(v)beta2, the LTCC chaperone for Ca(v)alpha1, antagonizes Ca(v)alpha1 protein degradation by preventing Ca(v)alpha1 PEST sequence recognition, leading to increased LTCC density and the consequent modulation of Ca(2+) channel function. This novel mechanism by which Akt modulates LTCC stability could profoundly influence cardiac myocyte Ca(2+) entry, Ca(2+) handling, and contractility. PMID- 19307603 TI - Coordinated regulation of heterochromatic genes in Drosophila melanogaster males. AB - Dosage compensation modifies the chromatin of X-linked genes to assure equivalent expression in sexes with unequal X chromosome dosage. In Drosophila dosage compensation is achieved by increasing expression from the male X chromosome. The ribonucleoprotein dosage compensation complex (DCC) binds hundreds of sites along the X chromosome and modifies chromatin to facilitate transcription. Loss of roX RNA, an essential component of the DCC, reduces expression from X-linked genes. Surprisingly, loss of roX RNA also reduces expression from genes situated in proximal heterochromatin and on the small, heterochromatic fourth chromosome. Mutation of some, but not all, of the genes encoding DCC proteins produces a similar effect. Reduction of roX function suppresses position effect variegation (PEV), revealing functional alteration in heterochromatin. The effects of roX mutations on heterochromatic gene expression and PEV are limited to males. A sex limited role for the roX RNAs in autosomal gene expression was unexpected. We propose that this reflects a difference in the heterochromatin of males and females, which serves to accommodate the heterochromatic Y chromosome present in the male nucleus. roX transcripts may thus participate in two distinct regulatory systems that have evolved in response to highly differentiated sex chromosomes: compensation of X-linked gene dosage and modulation of heterochromatin. PMID- 19307604 TI - Minimal effect of ectopic gene conversion among recent duplicates in four mammalian genomes. AB - Gene conversion between duplicated genes has been implicated in homogenization of gene families and reassortment of variation among paralogs. If conversion is common, this process could lead to errors in gene tree inference and subsequent overestimation of rates of gene duplication. After performing simulations to assess our power to detect gene conversion events, we determined rates of conversion among young, lineage-specific gene duplicates in four mammal species: human, rhesus macaque, mouse, and rat. Gene conversion rates (number of conversion events/number of gene pairs) among young duplicates range from 8.3% in macaque to 18.96% in rat, including a 5% false-positive rate. For all lineages, only 1-3% of the total amount of sequence examined was converted. There is no increase in GC content in conversion tracts compared to flanking regions of the same genes nor in conversion tracts compared to the same region in nonconverted gene-family members, suggesting that ectopic gene conversion does not significantly alter nucleotide composition in these duplicates. While the majority of gene duplicate pairs reside on different chromosomes in mammalian genomes, the majority of gene conversion events occur between duplicates on the same chromosome, even after controlling for divergence between duplicates. Among intrachromosomal duplicates, however, there is no correlation between the probability of conversion and physical distance between duplicates after controlling for divergence. Finally, we use a novel method to show that at most 5 10% of all gene trees involving young duplicates are likely to be incorrect due to gene conversion. We conclude that gene conversion has had only a small effect on mammalian genomes and gene duplicate evolution in general. PMID- 19307605 TI - Identification of EMS-induced mutations in Drosophila melanogaster by whole genome sequencing. AB - Next-generation methods for rapid whole-genome sequencing enable the identification of single-base-pair mutations in Drosophila by comparing a chromosome bearing a new mutation to the unmutagenized sequence. To validate this approach, we sought to identify the molecular lesion responsible for a recessive EMS-induced mutation affecting egg shell morphology by using Illumina next generation sequencing. After obtaining sufficient sequence from larvae that were homozygous for either wild-type or mutant chromosomes, we obtained high-quality reads for base pairs composing approximately 70% of the third chromosome of both DNA samples. We verified 103 single-base-pair changes between the two chromosomes. Nine changes were nonsynonymous mutations and two were nonsense mutations. One nonsense mutation was in a gene, encore, whose mutations produce an egg shell phenotype also observed in progeny of homozygous mutant mothers. Complementation analysis revealed that the chromosome carried a new functional allele of encore, demonstrating that one round of next-generation sequencing can identify the causative lesion for a phenotype of interest. This new method of whole-genome sequencing represents great promise for mutant mapping in flies, potentially replacing conventional methods. PMID- 19307606 TI - Translocation and assembly of mitochondrially coded Saccharomyces cerevisiae cytochrome c oxidase subunit Cox2 by Oxa1 and Yme1 in the absence of Cox18. AB - Members of the Oxa1/YidC/Alb3 family of protein translocases are essential for assembly of energy-transducing membrane complexes. In Saccharomyces cerevisiae, Oxa1 and its paralog, Cox18, are required for assembly of Cox2, a mitochondrially encoded subunit of cytochrome c oxidase. Oxa1 is known to be required for cotranslational export of the Cox2 N-terminal domain across the inner mitochondrial membrane, while Cox18 is known to be required for post translational export of the Cox2 C-tail domain. We find that overexpression of Oxa1 does not compensate for the absence of Cox18 at the level of respiratory growth. However, it does promote some translocation of the Cox2 C-tail domain across the inner membrane and causes increased accumulation of Cox2, which remains unassembled. This result suggests that Cox18 not only translocates the C tail, but also must deliver it in a distinct state competent for cytochrome oxidase assembly. We identified respiring mutants from a cox18Delta strain overexpressing OXA1, whose respiratory growth requires overexpression of OXA1. The recessive nuclear mutations allow some assembly of Cox2 into cytochrome c oxidase. After failing to identify these mutations by methods based on transformation, we successfully located them to MGR1 and MGR3 by comparative hybridization to whole-genome tiling arrays and microarray-assisted bulk segregant analysis followed by linkage mapping. While Mgr1 and Mgr3 are known to associate with the Yme1 mitochondrial inner membrane i-AAA protease and to participate in membrane protein degradation, their absence does not appear to stabilize Cox2 under these conditions. Instead, Yme1 probably chaperones the folding and/or assembly of Oxa1-exported Cox2 in the absence of Mrg1 or Mgr3, since respiratory growth and cytochrome c oxidase assembly in a cox18 mgr3 double mutant strain overexpressing OXA1 is YME1 dependent. PMID- 19307607 TI - Detection of protein-protein interactions through vesicle targeting. AB - The detection of protein-protein interactions through two-hybrid assays has revolutionized our understanding of biology. The remarkable impact of two-hybrid assay platforms derives from their speed, simplicity, and broad applicability. Yet for many organisms, the need to express test proteins in Saccharomyces cerevisiae or Escherichia coli presents a substantial barrier because variations in codon specificity or bias may result in aberrant protein expression. In particular, nonstandard genetic codes are characteristic of several eukaryotic pathogens, for which there are currently no genetically based systems for detection of protein-protein interactions. We have developed a protein-protein interaction assay that is carried out in native host cells by using GFP as the only foreign protein moiety, thus circumventing these problems. We show that interaction can be detected between two protein pairs in both the model yeast S. cerevisiae and the fungal pathogen Candida albicans. We use computational analysis of microscopic images to provide a quantitative and automated assessment of confidence. PMID- 19307608 TI - Quantifying interactions within the NADP(H) enzyme network in Drosophila melanogaster. AB - In this report, we use synthetic, activity-variant alleles in Drosophila melanogaster to quantify interactions across the enzyme network that reduces nicotinamide adenine dinucleotide phosphate (NADP) to NADPH. We examine the effects of large-scale variation in isocitrate dehydrogenase (IDH) or glucose-6 phosphate dehydrogenase (G6PD) activity in a single genetic background and of smaller-scale variation in IDH, G6PD, and malic enzyme across 10 different genetic backgrounds. We find significant interactions among all three enzymes in adults; changes in the activity of any one source of a reduced cofactor generally result in changes in the other two, although the magnitude and directionality of change differs depending on the gene and the genetic background. Observed interactions are presumably through cellular mechanisms that maintain a homeostatic balance of NADPH/NADP, and the magnitude of change in response to modification of one source of reduced cofactor likely reflects the relative contribution of that enzyme to the cofactor pool. Our results suggest that malic enzyme makes the largest single contribution to the NADPH pool, consistent with the results from earlier experiments in larval D. melanogaster using naturally occurring alleles. The interactions between all three enzymes indicate functional interdependence and underscore the importance of examining enzymes as components of a network. PMID- 19307609 TI - Selection for chaperone-like mediated genetic robustness at low mutation rate: impact of drift, epistasis and complexity. AB - Genetic robustness is defined as the constancy of a phenotype in the face of deleterious mutations. Overexpression of chaperones, to assist the folding of proteins carrying deleterious mutations, is so far one of the most accepted molecular mechanisms enhancing genetic robustness. Most theories on the evolution of robustness have focused on the implications of high mutation rate. Here we show that genetic drift, which is modulated by population size, organism complexity, and epistasis, can be a sufficient force to select for chaperone mediated genetic robustness. Using an exact analytical solution, we also show that selection for costly genetic robustness leads to a paradox: the decrease of population fitness on long timescales and the long-term dependency on robustness mechanisms. We suggest that selection for genetic robustness could be universal and not restricted to high mutation rate organisms such as RNA viruses. The evolution of the endosymbiont Buchnera illustrates this selection mechanism and its paradox: the increased dependency on chaperones mediating genetic robustness. Our model explains why most chaperones might have become essential even in optimal growth conditions. PMID- 19307610 TI - Leukocyte elastase induces lung epithelial apoptosis via a PAR-1-, NF-kappaB-, and p53-dependent pathway. AB - Leukocyte elastase induces apoptosis of lung epithelial cells via alterations in mitochondrial permeability, but the signaling pathways regulating this response remain uncertain. Here we investigated the involvement of proteinase-activated receptor-1 (PAR-1), the transcription factor NF-kappaB, and the protooncogene p53 in this pathway. Elastase-induced apoptosis of lung epithelial cells correlated temporally with activation of NF-kappaB, phosphorylation, and nuclear translocation of p53, increased p53 up-regulated modulator of apoptosis (PUMA) expression, and mitochondrial translocation of Bax resulting in enhanced permeability. Elastase-induced apoptosis was also prevented by pharmacologic inhibitors of NF-kappaB and p53 and by short interfering RNA knockdown of PAR-1, p53, or PUMA. These inhibitors prevented elastase-induced PUMA expression, mitochondrial translocation of Bax, increased mitochondrial permeability, and attenuated apoptosis. NF-kappaB inhibitors also reduced p53 phosphorylation. We conclude that elastase-induced apoptosis of lung epithelial cells is mediated by a PAR-1-triggered pathway involving activation of NF-kappaB and p53, and a PUMA- and Bax-dependent increase in mitochondrial permeability leading to activation of distal caspases. Further, p53 contributes to elastase-induced apoptosis by both transcriptional and post-transcriptional mechanisms. PMID- 19307612 TI - Journal of dairy science 2008 editorial report. PMID- 19307611 TI - Absence of proteinase-activated receptor-1 signaling in mice confers protection from fMLP-induced goblet cell metaplasia. AB - The morphological features of chronic obstructive pulmonary disease in man include emphysema and chronic bronchitis associated with mucus hypersecretion. These alterations can be induced in mice by a single intratracheal instillation of N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), a chemoattractant and degranulating agent for neutrophils. The mechanisms underlying excessive mucus production and, in particular, goblet cell hyperplasia/metaplasia in chronic obstructive pulmonary disease remain poorly understood. The proteinase-activated receptors (PARs) are widely recognized for their modulatory properties during inflammation. In this study, we examined whether PAR-1 contributes to inflammation and lung damage induced by fMLP by comparing the response of PAR-1 deficient (PAR-1(-/-)) mice with that of wild-type (WT) mice. Mice were killed at various time points after fMLP instillation (200 microg/50 microl). WT mice developed emphysema and goblet cell metaplasia. The onset of pulmonary lesions was preceded by an increase in thrombin immunoreactivity in bronchial airways and alveolar tissue. This was followed by a decrease in PAR-1 immunoreactivity, and by an increase in IL-13 immunostaining on the luminal surface of airway epithelial cells. In PAR-1(-/-) mice, fMLP administration induced similar responses in terms of inflammation and emphysema, but these mice were protected from the development of goblet cell metaplasia. The involvement of PAR-1 in airway epithelial cell transdifferentiation was confirmed by demonstrating that intratracheal instillation of the selective PAR-1 agonist (TFLLR) induced goblet cell metaplasia in the airways of WT mice only. These data suggest that emphysema and goblet cell metaplasia occur independently, and that PAR-1 signaling through IL-13 stimulation may play an important role in inducing goblet cell metaplasia. PMID- 19307613 TI - Occurrence of beta-casein fragments in cold-stored and curdled river buffalo (Bubalus bubalis L.) milk. AB - The safeguard of river buffalo Mozzarella cheese, a Protected Designation of Origin dairy product, has prompted an analytical study to trace the milk and curd used as raw material in cheesemaking. This is to prevent the illegal use of milk or curd from different geographical areas outside of those indicated in the official production protocol. For this purpose, we studied primary proteolysis occurring in fresh and frozen milk and curd to identify a molecular marker that could indicate the raw material used. Whole casein from frozen river buffalo milk was separated using cation-exchange chromatography and sodium dodecyl sulfate PAGE, and a protein component with an estimated molecular weight of 15.3 kDa was detected. This protein component was revealed in fresh river buffalo milk as a faint electrophoresis band, which drastically increased in intensity in refrigerated and frozen milk as well as in curd and was found to be associated with beta-CN through hydrophobic interaction. By using matrix-assisted laser desorption/ionization-time of flight peptide mass mapping, this component was identified as the C-terminal fragment f(69-209) of beta-CN (expected molecular weight of 15,748.8 Da). beta-Casein f(69-209), originating from the early hydrolysis of Lys(68)-Ser(69) by plasmin, has no counterpart in bovine milk. The increased rate of hydrolysis by plasmin toward the cleavage site Lys(68)-Ser(69) has to be ascribed to the elevated proline content of the peptide 61-73. The favored production of beta-CN f(69-209) has also drawn attention to the complementary proteose peptone beta-CN f(1-68) that is presumed to play a physiological role in inducing milk secretion similar to that of beta-CN f(1-29). The higher in vivo and in vitro production rate, compared with gamma(1)-CN formation, indicates that beta-CN f(69-209) and its complementary fragment are candidate molecular markers to evaluate milk and curd freshness. We suggested [corrected] indirect ELISA analysis based on the determination of remaining nonhydrolyzed beta-CN to perform a quantitative evaluation of proteolysis. PMID- 19307614 TI - Probiotic in lamb rennet paste enhances rennet lipolytic activity, and conjugated linoleic acid and linoleic acid content in Pecorino cheese. AB - Cheeses manufactured using traditional lamb rennet paste, lamb rennet paste containing Lactobacillus acidophilus, and lamb rennet paste containing a mix of Bifidobacterium lactis and Bifidobacterium longum were characterized for the lipolytic pattern during ripening. Lipase activity of lamb rennet paste, lamb rennet containing Lb. acidophilus, and lamb rennet containing a mix of bifidobacteria was measured in sheep milk cream substrate. Rennet paste containing probiotics showed a lipase activity 2-fold greater than that displayed by traditional rennet. Total free fatty acid (FFA) in sheep milk cream was lower in lamb rennet paste (981 microg/g of milk cream) than in lamb rennet containing Lb. acidophilus (1,382.4 microg/g of milk cream) and in lamb rennet containing a mix of bifidobacteria (1,227.5 microg/g of milk cream) according to lipase activity of lamb rennet paste. The major increase of FFA in all cheeses occurred during the first 30 d of ripening with the greatest values being observed for C16:0, C18:0 C18:1. At 60 d of ripening all cheeses showed a reduction in the amount of free fatty acids; in particular, total free fatty acids underwent a decrease of more than 30% from 30 to 60 d in cheeses manufactured using traditional lamb rennet paste, whereas the same parameter decreased 10% in cheeses manufactured using lamb rennet paste containing Lb. acidophilus and cheeses manufactured using lamb rennet paste containing a mix of B. lactis and B. longum. Cheese containing Lb. acidophilus was characterized by the greatest levels of total conjugated linoleic acids (CLA) 9-cis, 11-trans CLA and 9-trans, 11-trans CLA, whereas cheese containing bifidobacteria displayed the greatest levels of free linoleic acid. Rennet pastes containing viable cells of Lb. acidophilus and a mix of B. lactis and B. longum were able to influence the amount of FFA and CLA in Pecorino cheese during ripening. PMID- 19307615 TI - Review of the chemistry of alphaS2-casein and the generation of a homologous molecular model to explain its properties. AB - alpha(S2)-Casein (alpha(S2)-CN) comprises up to 10% of the casein fraction in bovine milk. The role of alpha(S2)-CN in casein micelles has not been studied in detail in part because of a lack of structural information on the molecule. Interest in the utilization of this molecule in dairy products and nutrition has been renewed by work in 3 areas: biological activity via potentially biologically active peptides, functionality in cheeses and products, and nutrition in terms of calcium uptake. To help clarify the behavior of alpha(S2)-CN in its structure function relationships in milk and its possible applications in dairy products, this paper reviews the chemistry of the protein and presents a working 3 dimensional molecular model for this casein. The model was produced by threading the backbone sequence of the protein onto a homologous protein: chloride intracellular channel protein-4. Overall, the model is in good agreement with experimental data for the protein, although the amount of helix may be over predicted. The model, however, offers a unique view of the highly positive C terminal portion of the molecule as a surface-accessible area. This region may be the site for interactions with kappa-carrageenan, phosphate, and other anions. In addition, most of the physiologically active peptides isolated from alpha(S2)-CN occur in this region. This structure should be viewed as a working model that can be changed as more precise experimental data are obtained. PMID- 19307616 TI - Rapid detection of bovine milk in yak milk using a polymerase chain reaction technique. AB - Yak milk contains a greater percentage of protein and has better quality than bovine milk. There has been an increasing focus on yak milk and milk products during the last few years. In the present study, a PCR-based assay was developed for the specific identification of bovine milk in yak milk by designing 3 primers targeting the mitochondrial ND1 gene. The use of 3 primers in a single PCR reaction set yielded 2 amplification fragments of 293 and 190 bp from bovine milk DNA, whereas only 1 amplification fragment of 293 bp was obtained in yak milk DNA. The technique was applied to raw and heat-treated binary mixtures of yak and bovine milks and enabled the specific detection of bovine milk with a detection limit of 0.1%. The assay developed is sensitive, fast, and straightforward, and it might be useful in the quality control of yak milk and milk products. PMID- 19307617 TI - Efficiency of serum protein removal from skim milk with ceramic and polymeric membranes at 50 degrees C. AB - Raw milk (2,710 kg) was separated at 4 degrees C, the skim milk was pasteurized (72 degrees C, 16 s), split into 3 batches, and microfiltered using pilot-scale ceramic uniform transmembrane pressure (UTP; Membralox model EP1940GL0.1microA, 0.1 microm alumina, Pall Corp., East Hills, NY), ceramic graded permeability (GP; Membralox model EP1940GL0.1microAGP1020, 0.1 microm alumina, Pall Corp.), and polymeric spiral-wound (SW; model FG7838-OS0x-S, 0.3 microm polyvinylidene fluoride, Parker-Hannifin, Process Advanced Filtration Division, Tell City, IN) membranes. There were differences in flux among ceramic UTP, ceramic GP, and polymeric SW microfiltration membranes (54.08, 71.79, and 16.21 kg/m2 per hour, respectively) when processing skim milk at 50 degrees C in a continuous bleed-and feed 3x process. These differences in flux among the membranes would influence the amount of membrane surface area required to process a given volume of milk in a given time. Further work is needed to determine if these differences in flux are maintained over longer processing times. The true protein contents of the microfiltration permeates from UTP and GP membranes were higher than from SW membranes (0.57, 0.56, and 0.38%, respectively). Sodium-dodecyl-sulfate-PAGE gels for permeates revealed a higher casein proportion in GP and SW permeate than in UTP permeate, with the highest passage of casein through the GP membrane under the operational conditions used in this study. The slight cloudiness of the permeates produced using the GP and SW systems may have been due to the presence of a small amount of casein, which may present an obstacle in their use in applications when clarity is an important functional characteristic. More beta lactoglobulin passed through the ceramic membranes than through the polymeric membrane. The efficiency of removal of serum proteins in a continuous bleed-and feed 3x process at 50 degrees C was 64.40% for UTP, 61.04% for GP, and 38.62% for SW microfiltration membranes. The SW polymeric membranes had a much higher rejection of serum proteins than did the ceramic membranes, consistent with the sodium-dodecyl-sulfate PAGE data. Multiple stages and diafiltration would be required to produce a 60 to 65% serum protein reduced micellar casein concentrate with SW membranes, whereas only one stage would be needed for the ceramic membranes used in this study. PMID- 19307618 TI - Increasing omega fatty acid content in cow's milk through diet manipulation: effect on milk flavor. AB - Milk with an increased content of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and conjugated linoleic acid (CLA) was obtained by incorporating fish oil into the feed of cows. The 4 feed treatments used were a control diet of 57% forage and 43% concentrate mix with EnerGII fat supplement at 1.65% of dietary DM, or EnerGII in the basal diet was partially replaced with 1) 0.21% partially ruminally inert calcium salts of 71% fish oil given at 0.41% of DM; 2) 0.41% inert calcium salts of 71% fish oil given at 0.83% of DM; or 3) 0.83% inert calcium salts of 43% fish oil given at 0.83% of DM. The cows were milked after 5 and 8 wk and the EPA, DHA, and CLA contents in the pasteurized whole milk were determined. The presence of off-flavors in the milk was investigated after 3 and 10 d of storage. Twelve judges were trained to evaluate the presence of grassy, fishy, oily, oxidized, and rancid off-flavors. Although levels of EPA, DHA, vaccenic acid, and CLA increased for all 4 treatments, a trained sensory panel detected no difference in milk flavor between treatments and the control, with little or no intensity of off-flavors. Results suggest that feeding fish oil and EnerGII at varying levels enhanced CLA, EPA, DHA, and total n-3 fatty acids in milk over the length of the experiment without negatively affecting milk flavor. This creates the potential for a more marketable and healthful product. PMID- 19307619 TI - Functional properties of whey proteins affected by heat treatment and hydrodynamic high-pressure shearing. AB - Two batches of native whey proteins (WP) were subjected to microfluidization or heat denaturation accompanied by microfluidization, followed by spray drying. Powders were assessed for their solubility, heat stability, coagulation time, and emulsifying and foaming properties. Effects of denaturation and shearing were examined by particle size analysis, differential scanning calorimetry, reducing and nonreducing sodium dodecyl sulfate-PAGE, and size exclusion-HPLC. Heat treatment significantly decreased solubility, whereas the number of microfluidization passes markedly improved solubility. The combined effect of heat and pressure significantly increased heat coagulation time. Emulsifying activity index substantially increased upon heat denaturation and was further enhanced by microfluidization. Emulsion stability appeared unaffected by the combined treatment, but the concentration of adsorbed protein on fat droplets was significantly increased. Foaming properties were diminished by heating. Particle size distribution patterns, sodium dodecyl sulfate-PAGE, and size exclusion-HPLC revealed disappearance of major WP and creation of relatively higher, as well as smaller, molecular weight aggregates as a result of the 2 treatments. The use of heat and microfluidization in combination could be used to stabilize WP against heat by producing microparticulated species that have different surface and colloidal properties compared with native WP. These results have implications for the use of WP as an additive in heat-processed foods. PMID- 19307620 TI - Short communication: can the mammopathogenic Escherichia coli P4 strain have a direct role on the caseinolysis of milk observed during bovine mastitis? AB - During bacterial bovine mastitis, the quality of milk is altered because of caseinolysis. Endogenous potential actors in milk responsible for this caseinolysis have been well studied, unlike the exogenous bacterial ones. The aim of this study was to evaluate the direct role in caseinolysis of a mammopathogenic strain, Escherichia coli P4. Secretion of at least 4 extracellular bacterial caseinolytic enzymes was highlighted by zymography, in 3 different growth media, and at each bacterial growth state, suggesting that their expression was constitutive. Different experimental conditions to evaluate caseinolytic potential did not show any significant caseinolytic activity of E. coli P4 and of the 4 extracellular proteases detected, suggesting that the high caseinolysis observed during E. coli bovine mastitis does result from endogenous milk actors. PMID- 19307621 TI - Herd-level risk factors for seven different foot lesions in Ontario Holstein cattle housed in tie stalls or free stalls. AB - Worldwide, there is considerable between-herd variation within individual foot lesion prevalence studies. This variation suggests that herd-level risk factors are important from a prevention perspective. The objective was to determine the effect of selected risk factors on the prevalence of 7 foot lesions in both tie stall and free-stall housing systems. As part of a cross-sectional foot lesion study 5 hoof trimmers recorded lesions for all cows that were foot trimmed in a herd. In addition, they completed a risk factor questionnaire for each herd. The impact of specific risk factors was evaluated using separate multi-variable models for both free-stall and tie-stall herds. The lesions evaluated were digital dermatitis, sole ulcer, sole hemorrhage, heel horn erosion, white line separations, white line abscess, and interdigital fibroma. Model types were selected based on herd-level lesion distribution. Detrimental risk factors identified in free-stall housing included increased alley scraping frequency (2.2 to 2.4-fold for sole ulcers) and trimming in summer or fall (-0.2-fold vs. spring and winter for digital dermatitis). Protective risk factors in free stalls included intermediate bedding depth (0.4-fold for 2.5 to 7.5 cm vs. more or less bedding for interdigital fibroma) and trimming heifers before calving (0.1-fold for white line abscess). In tie-stall herds no protective risk factors were identified. Detrimental risk factors for lesions in tie stalls included year round access to outside areas (2.1-fold increase in digital dermatitis, 3.5-fold for white line separation, and 7.0-fold for interdigital fibroma vs. no or only seasonal exercise access), routine spraying of feet (2.0-fold increase in digital dermatitis), larger herds (3.0-fold increase in interdigital fibroma vs. <41 cow herds), and the use of wood bedding material (6.5-fold vs. straw bedding for interdigital fibroma). The risk factors identified need further evaluation to determine the temporal relationships, as well as whether the relationships with foot lesions are causal. PMID- 19307622 TI - Effects of additional prostaglandin F2alpha and estradiol-17beta during Ovsynch in lactating dairy cows. AB - This study was designed to evaluate whether decreasing circulating progesterone (P4) or increasing circulating estradiol-17beta (E2) near the time of artificial insemination (AI) in an Ovsynch protocol would increase pregnancies per AI (P/AI) in lactating dairy cows. Six hundred nineteen lactating Holstein cows (n = 772 inseminations) received Ovsynch (GnRH-7 d-PGF(2alpha)-56 h-GnRH-16 h-timed AI). Cows were randomized in a 2 x 2 factorial experiment of 4 treatments to receive or not receive 25 mg of PGF(2alpha) 24 h after the standard PGF(2alpha) of Ovsynch, or 0.5 mg of E2 at the time of the final GnRH of Ovsynch, or both. Blood samples were collected 24 h after normal PGF(2alpha) and at final GnRH to evaluate circulating P4. Ovarian ultrasound was done at final GnRH to determine preovulatory follicle size. Ovulation was confirmed by ultrasound 5 d after AI. Treatment with additional PGF(2alpha) increased the percentage of cows that had complete luteal regression (95.6%) compared with control cows (84.6%). In contrast, additional PGF(2alpha) had no detectable effect on P/AI (control = 41.5% vs. + PGF(2alpha) = 44.7%). Supplementation with E2 increased expression of estrus (84.4 vs. 37.2%), but had no effect on overall fertility and even tended to have a negative effect on fertility in cows that ovulated to the second GnRH (control = 51.5% vs. +E2 = 44.0%). Thus, additional treatments with PGF(2alpha) or E2 during Ovsynch can be used to increase synchronization and expression of estrus during Ovsynch, although the lack of improvement in fertility makes these treatments unwarranted. PMID- 19307623 TI - Antibacterial effect of plant-derived antimicrobials on major bacterial mastitis pathogens in vitro. AB - The objective of this study was to investigate the antimicrobial effect of plant derived antimicrobials including trans-cinnamaldehyde (TC), eugenol, carvacrol, and thymol on major bacterial mastitis pathogens in milk. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the aforementioned compounds on Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli were determined. In addition, the bactericidal kinetics of TC on the aforementioned pathogens and the persistence of the antimicrobial activity of TC in milk over a period of 2 wk were investigated. All 4 plant-derived molecules exhibited antimicrobial activity against the 5 mastitis pathogens tested, but TC was most effective in killing the bacteria. The MIC and MBC of TC on Staph. aureus, E. coli, and Strep. uberis were 0.1 and 0.45%, respectively, whereas that on Strep. agalactiae and Strep. dysgalactiae were 0.05 and 0.4%, respectively. The MIC and MBC of the other 3 molecules ranged from 0.4 to 0.8% and 0.8 to 1.5%, respectively. In time-kill assays, TC at the MBC reduced the bacterial pathogens in milk by 4.0 to 5.0 log(10) cfu/mL and to undetectable levels within 12 and 24 h, respectively. The antimicrobial effect of TC persisted for the duration of the experiment (14 d) without any loss of activity. Results of this study suggest that TC has the potential to be evaluated as an alternative or adjunct to antibiotics as intramammary infusion to treat bovine mastitis. PMID- 19307624 TI - Effects of dietary energy and protein density on plasma concentrations of leptin and metabolic hormones in dairy heifers. AB - The hormonal and metabolic signals that communicate the level of body energy reserves to the reproductive-mammary axis remain undefined in dairy cattle; consequently, our hypothesis was that leptin may fulfill this role. Our objectives were to determine the effects of diets differing in energy and protein density on dry matter intake (DMI), growth traits [body weight (BW), body condition score (BCS), back-fat (BF) thickness], and temporal changes in plasma concentrations of leptin, insulin, growth hormone (GH), insulin-like growth factor-1 (IGF-1), glucose, and nonesterified fatty acids (NEFA) in dairy heifers during the pre- and postpubertal periods. In period 1, heifers were randomly allotted (n = 10/diet) at 103 kg of BW to diets for a predicted average daily gain of 1.10 (high, H), 0.80 (medium, M), or 0.50 kg/d (low, L). Five heifers in each of the H and L groups were further studied during period 2, either at 12 mo of age (HA, LA) or at 330 kg of BW (HW, LW). The data provide evidence that 1) DMI (18%), BW (17%), and BF (5%) together explained 40% of the variation in plasma leptin concentrations (r(2) = 0.396); 2) unlike the acute postprandial increase in plasma insulin as a result of increased nutrient density (H 1.42 +/- 0.09, M 1.02 +/- 0.09, L 0.68 +/- 0.11 ng/mL), plasma leptin concentrations did not respond acutely with a distinct postprandial profile; 3) although plasma leptin concentrations increased with age, leptin at puberty did not differ among treatment groups (H 5.63 +/- 2.48, M 4.28 +/- 0.55, L 4.12 +/- 0.72 ng/mL) and there was no evidence of an abrupt transition in prepubertal plasma leptin concentrations; 4) plasma leptin concentrations may not be a critical trigger for puberty in rapidly growing heifers, but are apparently essential for puberty in heifers with normal or restricted growth rates; and 5) plasma concentrations of insulin (H 0.59 +/- 0.07, M 0.43 +/- 0.09, L 0.30 +/- 0.09 ng/mL), IGF-1 (H 151.08 +/- 16.47, L 82.51 +/- 17.47 ng/mL), and glucose (H 81.35 +/- 3.39, M 73.59 +/- 2.34, L 68.25 +/- 3.39 mg/dL) reflected nutrient density, whereas GH (H 1.82 +/- 0.23, L 5.87 +/- 0.45 ng/mL) and NEFA (H 209.54 +/- 50.83, L 234.93 +/- 48.97 microM) were inversely related to the plane of nutrition. Collectively, these data suggest that plasma concentrations of leptin may play a role in long term regulation of energy reserves and puberty in growing Holstein heifers. PMID- 19307625 TI - Mastitis-related subtypes of bovine Staphylococcus aureus are characterized by different clinical properties. AB - Based on a former study from our group, one subtype of Staphylococcus aureus was associated with high within-herd prevalence of mastitis, whereas the other subtypes were associated with a low prevalence (sporadic intramammary infection). To confirm this hypothesis, a prospective study was done in 29 Swiss dairy herds. In particular, milk samples were collected from 10 herds with Staph. aureus herd problems (cases) and compared with samples from 19 herds with only sporadic cases of with Staph. aureus intramammary infection (controls). The isolates were tested for their virulence gene pattern and genotyped by PCR amplification of the 16S 23S rRNA intergenic spacer. The patterns and genotypes were then associated and compared with epidemiological and clinical data. Confirming the hypothesis, one particular subtype (genotype B) was associated with high within-herd and within cow prevalence of intramammary infection, whereas the other subtypes were associated with low within-herd prevalence and infected single quarters. The gene patterns and genotypes were highly related, demonstrating the genetic diversity of the genotypes. The somatic cell counts were clearly increased in herds with a genotype B problem compared with herds with infections of other genotypes. Based on the different clinical properties and treatment consequences associated with these different genotypes found in Switzerland, we recommend subtyping Staph. aureus in other countries to determine if this finding is universally applicable. PMID- 19307626 TI - Effects of milking interval and cisternal udder evaluation in Tunisian Maghrebi dairy dromedaries (Camelus dromedarius L.). AB - Effects of 4 different milking intervals (8, 12, 16, and 24 h) on milk yield and milk composition were studied in Tunisian Maghrebi dairy dromedaries (n = 6) at late lactation [240 +/- 14 days in milk (DIM), 5.84 +/- 1.62 L/d]. Camel-cows suckled their calves for 2 mo, were hand milked while suckling until mo 4 of lactation (calf weaning) and machine milked thereafter. Intravenous injection of oxytocin was administered before machine milking at each experimental milking to induce complete milk ejection and to avoid carryover effects of milking intervals. Cisternal and alveolar milk were measured at 380 +/- 16 DIM for a 24-h milking interval. Milk accumulated logarithmically (R(2) = 0.95) in the udder from 8- to 24-h milking interval without reaching a plateau. Consequently, milk secretion rate decreased exponentially (R(2) = 0.93) according to milking interval. Compared with 12-h milking interval (6.1 L/d), estimated daily milk yield was 113, 87, and 70% for 8-, 16-, and 24-h intervals, respectively. Total milk solids, milk fat content, and milk pH decreased with increasing milking interval, showing the greatest value at 8-h intervals (14.1 +/- 0.4%, 4.6 +/- 0.5%, and 6.66 +/- 0.05, respectively) and the lowest at 24-h intervals (12.3 +/- 0.9%, 2.9 +/- 0.6%, and 6.54 +/- 0.02, respectively). Milk protein (3.9 +/- 0.1%), lactose (4.5 +/- 0.2%), ash (0.84 +/- 0.01%) and density (1.028 +/- 0.01) remained constant for all milking intervals. Milk K, Ca, and Mg contents increased as milking interval increased, but Na content did not change (0.06 +/- 0.01%, on average). Milk Na:K ratio tended to decrease from 0.35 (1:2.9) to 0.22 (1:4.5) for the extreme milking intervals. Plasma lactose concentration steadied from 8- to 16-h (67 +/- 32 micromol) but increased dramatically at 24-h intervals (338 +/- 118 micromol), indicating that mammary tight junctions became permeable after 24 h of milk accumulation. Camel udders showed small cisterns (19.3% of total milk in the udder at 24 h) when compared with other dairy animals; we recommend the use of prestimulation for machine milking and selection for larger udder cisterns. Alveolar milk contained more fat (5.16 vs. 1.75%; SEM, 0.39%) and protein (3.23 vs. 2.73%; SEM, 0.15%) than cisternal milk. Despite the increase of plasma lactose during tight junction leakiness, the tendency for the Na:K ratio to decrease may be indicative of a camel's specific regulatory mechanism for controlling Na and K concentrations in milk and delaying the inhibitory effect of milk stasis on milk secretion rate. In conclusion, this short-term study proved the low storage capacity of the Tunisian Maghrebi camel udder but also showed their moderate ability to adapt to extended milking intervals at late lactation. PMID- 19307627 TI - Comparing subpopulations of plasma progesterone using cluster analyses. AB - The production of progesterone during the luteal phase of the estrous cycle regulates early embryonic development, uterine function, and luteal function. Plasma progesterone concentrations were measured during a spontaneous estrous cycle in lactating Holstein-Friesian cows of New Zealand or North American ancestry, fed either fresh pasture or a total mixed ration. Subpopulations of profiles were identified using 2 approaches: 1) shape-based clustering and 2) trait-based clustering. Subpopulations of profiles were compared using key progesterone profile components, which described the early and late luteal phase of the estrous cycle. The application of resistant nonlinear smoothing functions across raw progesterone profiles resulted in a high degree of agreement between the raw data and smoothed estimates (R(2) = 0.93). Both clustering techniques resulted in the identification of 3 similar subpopulations of progesterone profiles. The distribution of animals in these defined subpopulations and the association with cow strain and diet were examined. The early luteal phase was similar for the shape-based categories, but differed in the trait-based analyses. Differences in the late luteal phase and the onset of functional luteal regression in the different clusters were evident for both the shape- and trait based analyses. The distribution of animals across the subpopulations identified by shape-based clustering was not associated with either cow strain or diet. However, within the 3 subpopulations identified using trait-based clustering, cow strain affected the shape of the progesterone profile. Isolation of 3 subpopulations within a small data set showed that otherwise normal progesterone profiles exhibit significant variation. The differences in the luteal phase of these subpopulations, in particular progesterone early in the luteal phase and the early onset of functional luteal regression, may provide an insight into factors associated with subfertility. PMID- 19307628 TI - Effect of lactation stage and energy status on milk fat composition of Holstein Friesian cows. AB - The effects of lactation stage, negative energy balance (NEB), and milk fat depression (MFD) were estimated on detailed milk fat composition in primiparous Holstein-Friesian cows. One morning milk sample was collected from each of 1,933 cows from 398 commercial Dutch herds in winter 2005. Milk fat composition was measured using gas chromatography, and fat and protein percentage were measured using infrared spectrometry. Each fatty acid changed 0.5 to 1 phenotypic standard deviation over lactation, except odd-chain C5:0 to C15:0, branched-chain fatty acids, and trans-10, cis-12 conjugated linoleic acid (CLA). The greatest change was an increase from 31.2 to 33.3% (wt/wt) for C16:0 from d 80 to 150 of lactation. Energy status was estimated for each cow as the deviation from each average lactation fat-to-protein ratio (FPdev). A high FPdev (>0.12) indicated NEB. Negative energy balance was associated with an increase in C16:0 (0.696 +/- 0.178) and C18:0 (0.467 +/- 0.093), which suggested mobilization of body fat reserves. Furthermore, NEB was associated with a decrease in odd-chain C5:0 to C15:0 (-0.084 +/- 0.020), which might reflect a reduced allocation of C3 components to milk fat synthesis. A low FPdev indicated MFD (<-0.12) and was associated with a decrease in C16:0 (-0.681 +/- 0.255) and C18:0 (-0.128 +/- 0.135) and an increase in total unsaturated fatty acids (0.523 +/- 0.227). The study showed that both lactation stage and energy balance significantly contribute to variation in milk fat composition and alter the activity of different fatty acid pathways. PMID- 19307629 TI - Effect of type of diet and energy intake on milk production of Holstein-Friesian cows with extended lactations. AB - The aim of this study was to measure the effect of type of diet and level of energy intake on the performance of cows undergoing extended lactations. Ninety six Holstein-Friesian cows that calved in July and August 2004 were assigned randomly to 1 of 8 groups each of 12 cows (including 4 primiparous cows). Two of the 8 groups were assigned to each of 4 treatments that varied in lactation length (300 or 670 d) and diet (3 diets: control, high, or full total mixed ration (TMR). The 4 treatments were 1) control 300: cows were managed for a 300-d lactation and grazed pasture supplemented with grain and forage to provide a minimum daily dietary intake of 160 MJ of ME/cow; 2) control 670: as for control 300 except that cows were managed for a 670-d lactation; 3) high 670: cows were managed for a 670-d lactation and pasture was supplemented with grain and forage to provide a minimum daily dietary intake of 180 MJ of ME/cow; 4) full TMR 670: cows were managed for a TMR system that included a high body condition score at calving with cows offered a TMR during a 670-d lactation. The TMR was initially offered ad libitum indoors until about 440 DIM when the amount of TMR offered was reduced by about 2 kg of DM/d to prevent excessive weight gain. The proportions of cows still milking at the end of a 670-d lactation were similar for the control and high dietary groups. The full TMR group had fewer cows milking at 600 DIM: 17 cows milking compared with 24 cows in the control 670 group and 22 cows in the high 670 group. For the period 1 to 670 DIM, increasing the energy level in the diet (control 670 vs. high 670) resulted in a similar yield of milk and a similar fat concentration in the milk, but greater yields of milk fat and protein and greater milk protein percentage of the milk. The full TMR 670 group produced greater yields of milk and milk components (fat, protein, and lactose) and also protein percentage in the milk than the other groups. The milk solids (fat + protein) ratio for the 3 extended-lactation groups, defined as production achieved during the 24-mo calving interval divided by 2 yr (annualized production) expressed as a ratio of that produced in the normal 12-mo calving interval, was not affected by increasing the level of grain in the pasture-based diets (0.93 vs. 0.90 for control and high diets, respectively), but decreased with the TMR diet (0.79). The control 670 group produced 7.1% less milk, but only 2.4% less milk solids than the control 300 group over the 2-yr period of the study. Combining our data with that from 2 earlier studies of extended lactation demonstrated that Holstein cows with a high proportion of Northern Hemisphere genes offered pasture-based diets could achieve a high milk solids ratio, a greater proportion of cows milking at drying-off, and lower body weight gain over the lactation. PMID- 19307630 TI - Use of neural networks to detect minor and major pathogens that cause bovine mastitis. AB - The objectives of this research were to test the potential of unsupervised (USNN) and supervised neural network (SNN) models for detecting major and minor mastitis pathogens based on changes in milk parameters. A data set of 4,852 quarter milk samples with records for milk parameters and bacteriological status was used to train and validate the models by classifying milk samples into 3 different bacteriological states: not infected, intramammary infection (IMI) by minor pathogens, and IMI by major pathogens. Sensitivity of the USNN model was 97% for detecting noninfected quarters, 89% for minor pathogen IMI, and 80% for major pathogen IMI. Specificities of USNN models were close to 99% for all bacteriological states. The sensitivity of SNN models was affected by the ratio of infected to noninfected cases in the data set. As the ratio of infected to healthy cases increased from 1:1 to 1:10, detection accuracy for noninfected quarters increased from 82 to 98% but that for minor pathogen IMI decreased from 86 to 44%. The sensitivity for major pathogen IMI was 20% when the ratio was 1:1, but ranged from 20 to 40% when different ratios were tested. The SNN models indicated that somatic cell score and electrical resistance index had the most discriminating power. It was concluded that both USNN and SNN models were able to effectively differentiate between noninfected quarters and those infected by minor mastitis pathogens, and that the USNN model had a better agreement with results obtained from conventional microbiological methods. These types of models can be used in in-line milking systems to detect the infection status of a quarter and provide the farmer with diagnosing options for managing mastitis. PMID- 19307631 TI - Long-term performance of visual and electronic identification devices in dairy goats. AB - Dairy goat kids born during a 3-yr period (n = 97) and their mothers (n = 29) were used for a long-term evaluation of the performance of 9 types of identification (ID) devices. Kids wore multiple ID devices: visual ear tags (V1, tip-tag, n = 47; V2, official, n = 50), electronic ear tags (E1, button-button, n = 46; E2, flag-button, n = 46), electronic rumen boluses (B1, mini-bolus 14 g, n = 92; B2, mini-bolus 20 g, n = 28; B3, standard bolus 75 g, n = 34) and glass encapsulated transponders injected in the forefeet (T1, 15 mm, n = 75; T2, 12 mm, n = 100). Visual ear tags were applied at birth and removed in yearlings, whereas electronic ear tags were applied after bolusing with B1 (6.7 kg BW and 30 d, on average); B2 were administered in the event of a B1 loss, and B3 in case of a B2 loss and in goat does. At d 60 of age, kids were allocated into 2 groups to evaluate the effects of rearing system on ID. Treatments were: weaned (n = 46), and not weaned (n = 46) where kids suckled a milk substitute until d 150. Readability of ID devices (read/readable x 100) was monitored from 1 to 3 yr of age, depending on device and year of birth. Long-term readability was analyzed using a nonparametric survival analysis. A total of 3.3% infections and 6.5% tissue reactions were reported for electronic ear tags, but ears were fully healed in yearlings. Weaning numerically reduced B1 losses at d 150 (weaned, 84.8% vs. not weaned, 73.3%). Readability of visual ear tags in yearlings (V1, 82.9%; V2, 94.0%) was lower than for electronic ear tags (E1 and E2, 100%). Mini bolus readability in yearlings did not differ by type (B1, 71.4%; B2, 84.6%) or with visual ear tags. No effect of inject type was reported (T1, 92.0%; T2, 96.0%). Survival analysis after yr 3 gave the greatest readability value for E1 (100%), which did not differ from B3 (96.8%). The lowest readability was estimated for B1 (66.3%), followed by E2 (79.8%), B2 (81.4%), and T1 (90.4%). In conclusion, button-button electronic ear tags and standard boluses were the more efficient devices under our conditions, their readability values being greater than injects, electronic mini-boluses, and visual and flag-button electronic ear tags. Transponders injected in the forefeet and mini-boluses used here are not recommended in practice. Further research on E1 and B3 electronic devices should be done in a higher number of goats to confirm the current results. PMID- 19307632 TI - Effects of local anesthetic and a nonsteroidal antiinflammatory drug on pain responses of dairy calves to hot-iron dehorning. AB - This study examined the effects of a nonsteroidal antiinflammatory agent (NSAID) on physiological responses of calves immediately after hot-iron dehorning (DH) and during the time that local anesthetic (LA) wears off (2 to 3 h) after this procedure. Forty-six calves (33 +/- 0.3 d of age) were randomly assigned to 6 treatments: hot-iron DH versus sham DH with either no pain mitigation, LA alone, or LA with NSAID (i.v. Meloxicam). Eye temperature (measured using infrared thermography) was recorded every 5 min for 3 h after treatments. Heart rate (HR) and heart rate variability (HRV) were recorded continuously; for analysis of HRV, short segments of 512 interbeat intervals were examined. After DH without LA or NSAID, HR increased by 35 +/- 3.0 beats/min in the first 5 min and remained elevated above baseline for 3 h. The HRV around the time of DH did not differ between treatments; however, the root mean square of successive differences decreased from 68 to 41 +/- 12.6 ms immediately following DH without pain relief, suggesting a decrease in vagal tone at this time. Between 2 and 3 h following DH with LA, there was a decrease in eye temperature (-0.6 +/- 0.1 degrees C), an increase in HR (8 +/- 3.0 beats per min) and changes in HRV. Changes in HRV at this time included a decreased high-frequency power and an increase in the low frequency power and low-frequency/high-frequency ratio, indicating a change in sympatho-vagal balance. The changes in eye temperature, HR, and HRV between 2 and 3 h following DH with LA indicated the onset of pain coinciding with the time that the LA effects wear off. In addition, this study demonstrated that the combination of LA and NSAID mitigated the onset of pain responses when the LA wanes. PMID- 19307633 TI - Effect of fat source differing in fatty acid profile on metabolic parameters, fertilization, and embryo quality in high-producing dairy cows. AB - The objectives were to evaluate the effects of source of fatty acids (FA) on embryo quality of dairy cows. A total of 154 Holstein cows were assigned randomly to 1 of 2 sources of FA supplemented at 2% of the dietary dry matter as calcium salts of either palm oil (PO) or linoleic and trans-octadecenoic acids (LTFA) from 25 d prepartum to 80 d in milk (DIM). Cows were presynchronized beginning at 30 +/- 3 DIM and then subjected to the Ovsynch protocol beginning on d 39 +/- 3 postpartum. Timed artificial insemination was performed 12 h after the final GnRH of the Ovsynch protocol with semen from a single sire of proven fertility. The uteri of cows were nonsurgically flushed at 5 d after artificial insemination for collection of embryos-oocytes. Ovaries were examined by ultrasonography throughout the synchronization protocol. Blood was sampled and plasma was analyzed for concentrations of metabolites and hormones. The body condition score and yields of milk and milk components were measured throughout the first 90 DIM. Treatment did not affect concentrations of nonesterified FA, beta hydroxybutyrate, glucose, and progesterone in plasma. Body condition was similar between treatments. Milk production was similar between treatments, but concentrations of fat in milk and yields of fat and 3.5% fat-corrected milk decreased in cows fed LTFA, whereas concentration of true protein increased. Source of dietary FA did not influence ovulatory responses, diameter of the ovulatory follicle, and diameter of the corpus luteum during synchronization. Embryo-oocyte recovery relative to the number of corpora lutea did not differ between treatments. Fertilization tended to increase in cows fed LTFA compared with cows fed PO. Feeding LTFA improved the proportion of excellent-, good-, and fair-quality embryos, and embryos from cows fed LTFA had a greater number of blastomeres than embryos from cows fed PO. Feeding a more unsaturated source of FA improved fertilization and embryo development in lactating dairy cows, despite similar indicators of metabolic status. PMID- 19307634 TI - Effect of intrauterine infusion of ceftiofur on uterine health and fertility in dairy cows. AB - Objectives were to determine the effects of intrauterine (i.u.) infusion of ceftiofur hydrochloride on uterine health and fertility of dairy cows already receiving PGF(2alpha) for estrous synchronization. Holstein cows at 44 +/- 3 d in milk (DIM) were blocked by parity and diagnosis of metritis in the first 14 DIM and of other illnesses and, within each block, randomly assigned to 1 of 2 treatments: a single i.u. infusion of 125 mg of ceftiofur hydrochloride at 44 +/- 3 DIM (ceftiofur, n = 396), or no i.u. infusion (control, n = 416). All cows received 25 mg of PGF(2alpha) at 37 +/- 3 and 51 +/- 3 DIM as part of an estrous synchronization protocol. A subset of 547 cows was evaluated for clinical endometritis immediately before treatment, and 202 cows had an aseptic uterine sample collected before the injection of PGF(2alpha) at 51 +/- 3 DIM for bacteriology and diagnosis of subclinical endometritis (> or =5% neutrophils). Pregnancy on d 38 +/- 3 and 180 +/- 7 after the first artificial insemination, pregnancy loss, and interval from calving to pregnancy in the first 300 DIM were evaluated. The proportions of cows diagnosed with clinical endometritis before treatment were similar between ceftiofur and control treatments. Intrauterine infusion with ceftiofur did not influence prevalence of subclinical endometritis and positive uterine culture 7 d after treatment; however, it reduced the prevalence of positive uterine culture in cows with clinical endometritis (29.0 vs. 51.4%) and reduced the overall prevalence of Arcanobacterium pyogenes (1.0 vs. 7.6%) at 51 +/- 3 DIM. Cows with clinical endometritis had increased prevalence of A. pyogenes (10.3 vs. 1.5%), Escherichia coli (5.9 vs. 0.75%), and overall positive uterine culture (41.2 vs. 22.4%); however, cows with subclinical endometritis only had an increased prevalence of A. pyogenes (10.2 vs. 1.5%). Ceftiofur did not affect pregnancy per artificial insemination in all cows or in cows previously diagnosed with metritis or clinical endometritis. Interval to pregnancy was similar for control and ceftiofur cows. Intrauterine infusion of ceftiofur hydrochloride reduced the prevalence of uterine infection in cows with clinical endometritis, and the prevalence of A. pyogenes, but did not affect the prevalence of subclinical endometritis or fertility of dairy cows already receiving PGF(2alpha). PMID- 19307635 TI - Coordination of mammary metabolism and blood flow after refeeding in rats. AB - The production of milk is closely linked to nutritional state in many mammalian species, but the mechanisms by which changes in nutritional state are signaled to the mammary glands are poorly understood. Simultaneous measurements of mammary blood flow and glucose arterio-venous difference were made across the inguinal mammary glands of anesthetized, lactating rats. Blood flow to the mammary glands of previously fed rats was 0.48 mL/min per gram of mammary tissue. Glucose supply was 1.7 mumol/min per gram and 28% was extracted by the mammary glands. After food deprivation for 18 h, mammary blood flow decreased 48%, glucose arterio venous difference decreased 72%, and hematocrit increased 7%, resulting in a 60% decrease in glucose supply and an 88% decrease in glucose uptake. After 1 h of refeeding, glucose supply had returned to a similar level to that of normally fed animals, but glucose uptake was 60% higher than in the normally fed state. Mammary glucose uptake was not closely linked to either blood flow or glucose supply, suggesting that substrate supply was not the primary determinant of mammary metabolism. Denervation experiments showed that the mammary metabolic response to altered nutritional state was also unlikely to be closely controlled by neural pathways. Severance of the cutaneous branch of the posterior division of the femoral nerve innervating the inguinal mammary glands did not reduce the high glucose uptake by mammary glands of either fed or refed rats, nor did denervation change the low glucose uptake by mammary glands of food-deprived rats. Denervation reduced blood flow in the associated mammary gland, however, indicating that neural pathways may play a role in supporting mammary blood flow when food is available. In in vitro experiments, the rate of glucose uptake was 35% lower in mammary acini from food-deprived rats than in fed rats 2.5 h after tissue removal, indicating some persistence of the food deprivation-induced suppression of mammary metabolism. Administration of insulin increased glucose uptake in acini from both fed and food-deprived rats, indicating that insulin may be involved in signaling the mammary gland of the restoration of nutrient supply when food-deprived rats are refed. The effects of administration of a gut extract in vivo and in vitro are discussed. PMID- 19307636 TI - Performance and metabolic and endocrine changes with emphasis on glucose metabolism in high-yielding dairy cows with high and low fat content in liver after calving. AB - Elevated liver fat content occurs in high-yielding dairy cows during the transition from pregnancy to lactation after fat mobilization and may affect hepatic glucose metabolism, but the degree of liver fat storage is highly variable. Therefore, we studied metabolic and endocrine changes and hepatic glucose metabolism in cows that markedly differ in liver fat content. Multiparous cows from the same herd with high (HFL; n = 10) and low (LFL; n = 10) liver fat contents (mean of d 1, 10, and 21 after calving for each cow, respectively) were studied from 60 d before expected calving to 56 d in milk. Cows were fed ad libitum and all cows received the same diets. Liver samples were taken on d 1, 10, and 21 after calving; mean fat content (+/-SEM) in liver of HFL cows was 174 +/- 9.6 mg/g, whereas mean liver fat content in LFL cows was 77 +/- 3.3 mg/g. Blood samples were taken 20 and 7 d before expected calving and 0, 7, 14, 28, and 56 d after calving to measure plasma concentrations of nonesterified fatty acids, beta-hydroxybutyrate, glucose, insulin, glucagon, insulin-like growth factor-I, and leptin. In liver, glycogen content as well as mRNA levels of phosphoenolpyruvate carboxykinase, pyruvate carboxylase, glucose-6-phosphatase, and glucose transporter were measured by quantitative real-time PCR. Back fat thickness decreased and dry matter intake increased with onset of lactation, and back fat thickness was higher but dry matter intake was lower in HFL than in LFL. Energy-corrected milk yield did not differ between groups, but milk fat content was higher and lactose content was lower in HFL than LFL at the beginning of lactation. Energy balance was more negative in HFL than in LFL. Plasma nonesterified fatty acids and beta-hydroxybutyrate concentrations increased and plasma glucose concentration tended to decrease more in HFL than LFL with onset of lactation. Glucagon to insulin ratios increased more in HFL than LFL with onset of lactation. Hepatic glycogen content was higher in LFL than HFL, whereas mRNA levels of glucose-6-phosphatase and pyruvate carboxylase were higher in HFL than in LFL, and cytosolic phosphoenolpyruvate carboxykinase mRNA level increased similarly after parturition in both groups. In conclusion, an elevated liver fat content was related to greater fat mobilization and reduced feed intake and was associated with effects on hepatic glucose metabolism. As environment and feeding management were the same, individual cow factors were responsible for differences in energy metabolism during the transition period. PMID- 19307637 TI - Assessing lameness in cows kept in tie-stalls. AB - Identifying lame cows and quantifying the prevalence of lameness are important elements of cattle welfare assessment that are generally achieved by methods involving observations of each animal walking. There is no published method for assessing lameness in cows confined in tie-stalls. The objective of this study (carried out within the European Commission's Welfare Quality(R) project) was to develop a suitable method and validate it for lameness detection against a published locomotion score. A series of indicators of lameness visible in tied cows was formalized into a stall assessment protocol. This was validated against a traditional locomotion score and tested for repeatability between 2 observers. A total of 98 cows on 4 farms were assessed. Overall interobserver agreement was 91%. Sensitivity compared with locomotion scoring was 0.54 to 0.77, dependent on observer and threshold definition. Assessment in the stall underestimated the herd prevalence of lameness revealed by locomotion scoring by 11 to 37% (mean 27%). The discrepancy between herd lameness prevalence assessed in the stall and by locomotion scoring was not affected significantly by farm or observer. The cases of lameness that were not detected in the stall tended to be the least severe. The proposed method for lameness detection in tie-stalls could be used for herd-level assessment of lameness and detection of individual lame animals by farmers and their advisors, but it is important to remember that it is less sensitive than locomotion scoring. PMID- 19307638 TI - Effect of high somatic cell counts on reproductive performance of Chilean dairy cattle. AB - The objectives were to evaluate the effect of high linear somatic cell counts (LNSCC > or =4.5) during early lactation on reproductive performance and to estimate their association with the risk of abortion in a population of central southern Chilean dairy cattle. The analysis included records from a population of 157 farms and considered 1,127,405 test-day records including 101,944 lactations that began between 1997 and 2006. After data edits, the analyses of calving to first service and calving to conception intervals consisted of 88,633 and 70,877 lactations, respectively. Once controlling for significant variables, time to first breeding was 21.8 d longer in cows with at least 1 high LNSCC before the first breeding compared with controls. Cows with at least 1 high LNSCC before the fertile breeding had an increment in time to conception of 48.7 d and required, on average, 0.49 more services to conceive. The odds of conception at first service in cows with a high LNSCC within 30 d before [after] breeding were 0.85 (0.81 to 0.89; 95% confidence interval ) [0.82 (0.78 to 0.87; 95% confidence interval)] times the odds of conception for cows without a high LNSCC during that period. The Cox proportional hazard model indicated that after correction by calving year, lactation number, and milk yield standardized to 305 d, the risk of pregnancy decreased by 44% if a high LNSCC occurred before breeding. Cows registering a high LNSCC during the first 90 d of gestation had an increased risk of abortion, being 1.22 (1.07 to 1.35; 95% confidence interval) times more likely to abort than nonaffected cows. It is concluded that subclinical mastitis, measured as LNSCC >/=4.5, had a significant effect on reproductive performance in Chilean dairy cattle. PMID- 19307639 TI - Short communication: investigation of Coxiella burnetii occurrence in dairy sheep flocks by bulk-tank milk analysis and antibody level determination. AB - To estimate the prevalence of Coxiella burnetii in the dairy sheep population from the Basque Country (northern Spain), a study was carried out combining molecular and serological techniques. First, bulk-tank milk samples from 154 flocks belonging to the Latxa Breed Farmers Association were analyzed by PCR, with 22% of flocks testing positive for C. burnetii. Then, a selection of 34 flocks (7 PCR positive and 17 negative) was investigated for the presence of serum antibodies by ELISA test on 1,011 ewes (approximately 30 ewes per flock). A total of 8.9% of the animals were seropositive, 67.6% of the flocks had at least one seropositive animal, but only in 14.7% of them was seroprevalence greater than 25%. Older ewes showed a significantly greater prevalence (17.5%) compared with yearlings (7.5%) or replacement lambs (1.5%). A marginally significant association was found between seroprevalence and PCR detection of C. burnetii in bulk-tank milk. The widespread distribution of C. burnetii in the region advocates for the implementation of Q fever control strategies and highlights the potential risk of sheep as a reservoir and infection source for other domestic and wildlife species and the human population. PMID- 19307640 TI - Effects of feed naturally contaminated with Fusarium mycotoxins on metabolism and immunity of dairy cows. AB - A previous study in dairy cows showed some effect of feed contaminated with Fusarium mycotoxins on metabolism and immunity. A subsequent experiment investigated the effect of feedborne Fusarium mycotoxins on some immune functions in more detail. A total mixed ration (TMR) containing a blend of feedstuffs naturally contaminated with Fusarium mycotoxins was fed for 63 d to 12 mid lactation Holstein cows with an average milk production of 36 kg/d in a completely randomized design with repeated measures including 1) control TMR and 2) contaminated TMR. Wheat, corn, hay, and corn silage were the contaminated feedstuffs. Deoxynivalenol was the major contaminant and was found in TMR at 3.5 mg/kg of dry matter. The parameters measured were 1) performance: body weight, body condition score, dry matter intake, milk production, composition and somatic cell count; 2) health: blood serum chemistry, hematology, coagulation profile, and rumen fluid ammonia levels; 3) immune function: total serum immunoglobulins (IgA, IgG, IgM), specific antibody response to ovalbumin, and neutrophil phagocytosis. Dry matter intake, body weight, milk production, and milk composition were not affected by diet. Neutrophil phagocytosis was depressed throughout the experiment in cows fed the contaminated diet. Serum sodium concentrations and osmolality were significantly elevated throughout the experiment in cows fed the contaminated diet. Primary antibody response to ovalbumin immunization was higher in cows fed the contaminated diet compared with controls. It was concluded that feed naturally contaminated with Fusarium mycotoxins can affect metabolic parameters and immune function of dairy cows. PMID- 19307641 TI - Nutrient demand interacts with forage family to affect nitrogen digestion and utilization responses in dairy cows. AB - The effect of preliminary feed intake on responses to diets containing alfalfa silage or orchardgrass silage was evaluated using 8 ruminally and duodenally cannulated Holstein cows in a crossover design experiment with a 14-d preliminary period and two 15-d treatment periods. Responses measured were intake, digestion, and utilization of N. Cows were 139 +/- 83 (mean +/- standard deviation) days in milk at the beginning of the preliminary period. During the 14-d preliminary period, 3.5% fat-corrected milk yield ranged from 23.9 to 47.6 kg/d (mean = 36.9 kg/d) and preliminary voluntary dry matter intake (pVDMI) ranged from 14.2 to 21.3 kg/d (mean = 18.6 kg/d). Treatments were a diet with alfalfa silage as the sole forage (AL) and a diet with orchardgrass silage as the sole forage (OG). Alfalfa silage contained 20.5% crude protein (CP; dry matter basis) and orchardgrass silage contained 20.4% CP; AL contained 18.3% CP and 5.6% estimated rumen-undegraded CP, and OG contained 18.8% CP and 6.3% estimated rumen undegraded CP. Mean N intake was similar between treatments, ruminal N digestibility was greater for AL (30.4%) than for OG (17.7%), and whole-tract N digestibility did not differ between treatments. Intake and duodenal flow of N depended on a treatment x pVDMI interaction; both N intake and duodenal flow increased more for AL than for OG as pVDMI increased. Duodenal flow of microbial N and the efficiency of microbial N production from OM also depended on a treatment x pVDMI interaction in a manner similar to N intake and duodenal flow. However, treatment x pVDMI interactions also indicate that as pVDMI increased and N intake increased for AL compared with OG, a decreasing proportion of the additional N consumed from AL was digested and used for increased milk protein production or body tissue gain. Therefore, when feeding less-filling diets, such as those containing large proportions of legume forage, to high-producing cows, reducing dietary N concentration could increase the efficiency of N utilization and reduce the extent to which greater DMI leads to greater N excretion. PMID- 19307642 TI - Increasing physically effective fiber content of dairy cow diets through forage proportion versus forage chop length: chewing and ruminal pH. AB - A study was conducted to evaluate whether the risk of acidosis in dairy cows can be lowered by increasing the physically effective fiber (peNDF) concentration of the diet, either through increased theoretical chop length of alfalfa silage or higher proportion of forage in the diet. The experiment was designed as a replicated 4 x 4 Latin square using 8 ruminally cannulated lactating dairy cows. Treatments were arranged in a 2 x 2 factorial design; 2 forage particle lengths (FPL) of alfalfa silage (short and long) were combined with low (35:65) and high (60:40) forage:concentrate (F:C) ratios [dry matter (DM) basis]. Dietary peNDF concentration (DM basis) was determined from the sum of the proportion of dietary DM retained either on the 2 sieves (8 and 19 mm) or on the 3 sieves (1.18, 8, and 19 mm) of the Penn State Particle Separator multiplied by the neutral detergent fiber concentration of the diet. The dietary peNDF concentrations were altered by changing the F:C or the FPL, and ranged from 10.7 to 17.5% using 2 sieves, or from 23.1 to 28.2% using 3 sieves. Intake of peNDF was increased by increasing FPL but not by increasing F:C ratio because of the reduction of DM intake at the higher F:C ratio. Chewing activity, including number of chews and chewing time, increased with increasing F:C ratio or FPL. Mean ruminal pH was elevated by 0.4 and 0.2 units with increasing F:C ratio and FPL, respectively. Lowering the F:C ratio decreased the duration that ruminal pH was below 5.8 (1.2 vs. 8 h/d). Increased F:C ratio or FPL reduced ruminal volatile fatty acids concentration from 137 to 122 or from 133 to 126 mM, respectively, whereas acetate:propionate ratio was increased from 2.55 to 3.46 with increasing F:C ratio. Dietary peNDF concentration measured using 2 sieves was correlated to chewing time (r = 0.57) and mean ruminal pH (r = 0.75), whereas dietary peNDF concentration measured using 3 sieves was correlated to mean ruminal pH (r = 0.83) and negatively correlated to the time that pH was below 5.8 (r = -0.78). This study shows that the risk of ruminal acidosis is high for cows fed a low F:C diet. Increasing the proportion of forage in the diet helps prevent ruminal acidosis through increased chewing time, a change in meal patterns, and decreased ruminal acid production. Increasing FPL elevates ruminal pH, but in low forage diets, increased FPL does not alleviate subacute acidosis because the fermentability of the diet is high and changes in chewing activity are marginal. PMID- 19307643 TI - Effects of newborn characteristics and length of colostrum feeding period on passive immune transfer in goat kids. AB - Majorera goat kids (n = 200) were used to evaluate the effects of litter size, birth body weight, sex, and suckling duration on serum IgG concentrations. Kids were assigned to 1 of 3 experimental groups: litter size and sex were equally distributed in each group. In the first group, kids (n = 67) stayed with their dams for 24 h; in the second group, kids (n = 66) stayed with their dams for 48 h; and in the third group, kids (n = 67) stayed with their dams for 120 h. Blood samples were obtained every 24 h for 5 d, and serum IgG concentration was measured using radial immunodiffusion. In litter sizes of 1 to 2 kids, IgG blood serum concentration was significantly higher (18.30 +/- 5.40 mg/mL) than in litters of 3 kids (9.85 +/- 4.23 mg/mL). Kid sex did not affect IgG blood serum concentrations. Suckling duration did not affect kid serum IgG concentrations. In conclusion, kids with low birth body weight (<2.8 kg) or from litters of 3 may need special attention. If newborn goat kids are allowed to suckle colostrum for at least 24 h from their dams, this seems to be sufficient time to ingest enough IgG from colostrum to achieve an adequate serum IgG concentration and passive immune protection to avoid failure of passive immune transfer. PMID- 19307644 TI - Meta-analysis of the influence of Saccharomyces cerevisiae supplementation on ruminal parameters and milk production of ruminants. AB - The effects of yeast supplementation on intake, production, and rumen fermentation characteristics have been widely studied, but results are inconsistent between different studies. A quantitative meta-analysis was applied to 110 papers, 157 experiments, and 376 treatments dealing with yeast supplementation in ruminants. The objective was first to highlight the major quantitative effects of live yeast supplementation on intake, rumen fermentation, and milk production, and second, to identify major differences in experimental conditions between studies that can affect the response to treatment. Some of these experimental conditions are referred to as interfering factors. Yeast supplementation increased rumen pH (+0.03 on average) and rumen volatile fatty acid concentration (+2.17 mM on average), tended to decrease rumen lactic acid concentration (-0.9 mM on average), and had no influence on acetate-to-propionate ratio. Total-tract organic matter digestibility was also increased by yeast supplementation (+0.8% on average). Yeast supplementation increased dry matter intake (DMI; +0.44 g/kg of body weight; BW), milk yield (+1.2 g/kg of BW), and tended to increase milk fat content (+0.05%), but had no influence on milk protein content. Dose effects of yeast supplementation, expressed as log(10) [1+(cfu per 100 kg of BW)], globally confirmed the qualitative effects observed in the first analysis. The positive effect of yeast supplementation on rumen pH increased with the percentage of concentrate in the diet and with the DMI level. It was negatively correlated with the level of dietary neutral detergent fiber (NDF). The positive effect of yeast supplementation on rumen volatile fatty acid concentration increased with DMI and crude protein levels. The positive effect of yeast supplementation on organic matter digestibility increased with the percentage of concentrate and NDF in the diet. The negative effect of yeast supplementation on lactic acid concentration tended to decrease when the DMI level and the percentage of concentrate in the diet increased. The effects of interfering factors were globally similar when either dose effect or qualitative effect of yeast was taken into account. Although rumen fermentation efficiency per se was not measured, these results suggest an improvement in rumen fermentation by yeast supplementation. This effect could, however, be modulated by several different factors such as DMI, percentage of concentrate or NDF in the diet, or species. PMID- 19307645 TI - Effects of silage protein degradability and fermentation acids on metabolizable protein concentration: a meta-analysis of dairy cow production experiments. AB - A meta-analysis was conducted using data from dairy cow production studies to evaluate silage metabolizable protein (MP) concentrations. The data consisted of 397 treatment means in 130 comparisons, in which the effects of silage factors (e.g., date of harvest, wilting, silage additives) were investigated. Within a comparison, a fixed amount of the same concentrate was fed. A prerequisite of data to be included in the analysis was that silage dry matter (DM), crude protein (CP), ammonia N, lactic acid (LA), and total acid (TA) concentrations and digestibility were determined. A smaller data set (n = 248) comprised studies in which silage water-soluble N concentration was also analyzed. The supply of MP was estimated as amino acids absorbed from the small intestine using a model with constant values for ruminal effective protein degradability (EPD) and intestinal digestibility of rumen undegraded protein. Microbial protein was calculated on the basis of digestible carbohydrates and rumen degradable protein (RDP). Alternative models were used to estimate microbial protein formation, assuming the energy values of RDP and TA to be equivalent to 1.00, 0.75, 0.50, 0.25, and 0 times that of digestible carbohydrates. Because EPD values are seldom determined in production trials, they were derived using empirical models that estimate them from other feed components. The goodness of fit of models was compared on the basis of root mean squared error (RMSE) of milk protein yield (MPY) predicted from MP supply (adjusted for random study effect) and Akaike's information criterion. Metabolizable protein supply calculated from basal assumptions predicted MPY precisely within a study (RMSE = 16.2 g/d). Variable contribution of RDP to the energy supply for microbial synthesis influenced the precision of MPY prediction very little, but RMSE for MPY increased markedly when the energy supply of rumen microbes was corrected for TA concentration. Using predicted rather than constant EPD values also increased RMSE of MPY prediction. These observations do not mean that the supply of MP from undegraded feed protein is constant. However, it suggests that our current methods overestimate the range in EPD values and that the techniques have so many inherent technical problems that they can mask the true differences between the feeds. Including new elements in feed protein evaluation models may not improve the precision of production response predictions unless the consequent effects on the supply of other nutrients are taken into account. PMID- 19307646 TI - Effect of nitazoxanide on cryptosporidiosis in experimentally infected neonatal dairy calves. AB - Cryptosporidium is a zoonotic protozoan that is most often diagnosed in association with diarrhea in 1- to 3-wk-old dairy calves. There are neither consistently effective nor approved antimicrobial drugs for treatment in animals. The objective of this study was to test nitazoxanide (NTZ) as a treatment for cryptosporidiosis in experimentally infected dairy calves. A randomized, controlled, and blinded trial was performed using Holstein bull calves obtained from a large commercial dairy. All births were attended by study personnel and calves were fed 4 L of heat-treated colostrum within 1 h of birth. Calves were randomly assigned to treatment or placebo group and maintained for a 32-feeding (16 d) study period. Twenty-three calves were enrolled with 3 lost to follow up. Thirteen calves were assigned to the treatment group and 7 calves to the placebo group. All calves were inoculated with 1 x 10(6) viable Cryptosporidium parvum oocysts at feeding 3. Treatment was a commercially available NTZ product and the placebo was the carrier of the same product. Nitazoxanide was administered at 1.5 g twice per day for 5 d. Nitazoxanide or placebo treatment began after feeding 10 and when the fecal score was greater than 1 out of 3. Outcome measurements included twice-daily fecal and health scores and a once-daily oocyst count by an immunofluorescent antibody assay. Data were analyzed by nonparametric and time-to event methods. Measures of passive transfer of antibodies, initial body weight, and onset of oocyst shedding were not different between treatment and control calves. Eighty-five percent of the NTZ-treated calves stopped shedding oocysts by the end of the observation period whereas only 15% of the placebo group stopped shedding. The median number of feedings with a fecal score equal to 3 was 2 in the NTZ group while it was 6 in the placebo group. Calves receiving NTZ were 0.13 times as likely to have severe and sustained diarrhea than control calves (95% confidence interval, 0.02-0.98). Treating calves with NTZ reduced the duration of oocyst shedding and improved fecal consistency. PMID- 19307647 TI - Influence of different oral rehydration solutions on abomasal conditions and the acid-base status of suckling calves. AB - The aim of the study was to investigate the influence of oral rehydration solutions (ORS) on milk clotting, abomasal pH, electrolyte concentrations, and osmolality, as well as on the acid-base status in blood of suckling calves, as treatment with ORS is the most common therapy of diarrhea in calves to correct dehydration and metabolic acidosis. Oral rehydration solutions are suspected to inhibit abomasal clotting of milk; however, it is recommended to continue feeding cow's milk or milk replacer (MR) to diarrheic calves to prevent body weight losses. Three calves with abomasal cannulas were fed MR, MR-ORS mixtures, or water-ORS mixtures, respectively. Samples of abomasal fluid were taken before and after feeding at various time points, and pH, electrolyte concentrations, and osmolality were measured. The interference of ORS with milk clotting was examined in vivo and in vitro. To evaluate the effects of ORS on systemic acid-base status, the Stewart variables strong ion difference ([SID]), acid total ([A(tot)]), and partial pressure of CO2 (pCO2) were quantified in venous blood samples drawn before and after feeding. Calves reached higher abomasal pH values when fed with MR-ORS mixtures than when fed MR. Preprandial pH values were re established after 4 to 6 h. Oral rehydration solutions prepared in water increased the abomasal fluid pH only for 1 to 2 h. Oral rehydration solutions with high [SID(3)] ([Na(+)] + [K(+)] - [Cl(-)]) values produced significantly higher abomasal pH values and area under the curve data of the pH time course. Caseinomacropeptide, an indicator of successful enzymatic milk clotting, could be identified in every sample of abomasal fluid after feeding MR-ORS mixtures. The MR-ORS mixtures with [SID(3)] values > or =92 mmol/L increased serum [SID(3)] but did not change venous blood pH. Oral rehydration solutions do not interfere with milk clotting in the abomasum and can, therefore, be administered with milk. In this study, MR-ORS mixtures with high [SID(3)] values caused an increase of serum [SID(3)] in healthy suckling calves and may be an effective treatment for metabolic acidosis in calves suffering from diarrhea. PMID- 19307648 TI - Calf health monitoring in Norwegian dairy herds. AB - The aims of this study were to estimate the incidence of calf diseases in Norwegian dairy herds, improve calf health recordings in the Norwegian Cattle Health Recording System (NCHRS), and investigate different methods for validation of calf health data. A longitudinal, cross-sectional survey of calf health in Norway was performed between September 1, 2004, and January 31, 2007. The participating dairy herds were randomly selected from among herds registered in the Norwegian Dairy Herd Recording System as having at least 15 cow-years. Each herd participated for 1 yr. Diseases and treatments of calves of up to 180 d of age in 135 dairy herds were reported using the NCHRS. In total, 6,668 calves were born in the participating dairy herds during the project period. A total of 573 (29.6%) of the 1,936 calf health recordings reported were recordings of diseases and 1,363 (70.4%) were events of preventive therapy, dehorning, or castration. The recorded incidence of diarrhea and respiratory disease was 3.8% and 2.9%, respectively. The median age of occurrence of diarrhea and respiratory disease was 17 and 37 d, respectively. Three different methods, based on sampling of diseased calves, dehorning as an indicator of a well-functioning recording system, or feedback on degree of commitment to calf health recording, were tested to assess validation of the calf health records. The 3 methods indicated an underestimation of calf health records in the NCHRS of approximately 40% and an estimated "true" incidence of diarrhea and respiratory disease of 5.5 and 4.1%, respectively. The results from this study demonstrate the importance of encouraging farmers to conduct calf health recordings. They also indicate that finding a standardized method for validation of health data is a considerable challenge. PMID- 19307649 TI - Metabolism and lactation performance in dairy cows fed a diet containing rumen protected fat during the last twelve weeks of gestation. AB - Effects of dietary fat supplementation prepartum on liver lipids and metabolism in dairy cows are contradictory. Thus, we examined in 18 German Holstein cows (half-sib; first lactation 305-d milk yield >9,000 kg) whether dietary fat:carbohydrate ratio during the last trimester of gestation affects lipid metabolism and milk yield. The diets were formulated to be isoenergetic and isonitrogenous but differed in rumen-protected fat (FD; 28 and 46.5 g/kg of dry matter during far-off and close-up dry period; mainly C16:0 and C18:1) and starch concentration [carbohydrate diet (CD); 2.3 times as much starch as FD]. Diets were given ad libitum starting 12 wk before expected parturition. After parturition all cows were fed a single lactation diet ad libitum for 14 wk. With the FD treatment, dry matter intake was depressed prepartum, milk yield during first 4 wk of lactation was lower (36.9 vs. 41.0 kg/d), and postpartum energy balance during this period was more negative. During the first 4 wk, cows in the FD group had lower lactose percentage and yield but higher milk fat, whereas milk protein and fat yield as well as energy-corrected milk did not differ. Between wk 5 and 14, milk fat and milk protein percentage was lower in CD than in FD. Milk fat C14:0 was lower and C16:1 was higher in the FD group. For FD cows, plasma triacylglycerol, nonesterified fatty acids, and cholesterol concentrations were higher prepartum, whereas plasma beta-hydroxybutyrate and glucose concentrations were lower. During the first 10 d after parturition, plasma triacylglycerol concentration was higher in FD, and prepartum plasma glucose and cholesterol differences persisted during the first 14 wk of lactation. Irrespective of prepartum nutrient composition, concentrations of plasma leptin and subcutaneous fat leptin mRNA decreased between -10 d to +10 d relative to parturition, and liver lipids and glycogen reached maximum and minimal values, respectively, 10 d after parturition. Acetyl-coenzyme A carboxylase alpha mRNA abundance in subcutaneous fat decreased between -10 d to +1 d relative to parturition by 97%, whereas it was generally much lower in the liver and remained at a low level until wk 14 of lactation. In conclusion, feeding a diet containing rumen protected fat during late lactation and dry period until calving negatively affected dry matter intake, energy balance, and milk yield during subsequent lactation, did not change acetyl-coenzyme A carboxylase alpha mRNA abundance in subcutaneous fat, and was not beneficial for liver lipid accumulation. PMID- 19307650 TI - Influence of methionine supply on the response of lactational performance of dairy cows to supplementary folic acid and vitamin B12. AB - The present experiment was undertaken to determine if the effects of supplementary folic acid on lactational performance were caused by improved methylneogenesis and if the supply in vitamin B(12) could affect this metabolic pathway. In this eventuality, supplementary Met, a major source of preformed methyl groups, should reduce the requirements for these vitamins. Sixty multiparous Holstein cows were assigned to 10 blocks of 6 cows each according to their previous milk production. Within each block, 3 cows were fed a diet estimated to supply Met as 1.83% metabolizable protein and 3 cows were fed the same diet supplemented with 18 g of rumen-protected methionine (RPM) to supply Met as 2.23% of metabolizable protein. Within each level of Met, cows received no vitamin supplement or weekly intramuscular injections of 160 mg of folic acid alone or combined with 10 mg of vitamin B(12) from 3 wk before to 16 wk after calving. There was no treatment effect on dry matter intake during pre- and postcalving periods: 13.4 +/- 0.4 and 21.8 +/- 0.4 kg/d, respectively. Milk production was not affected by RPM supplementation. Folic acid and vitamin B(12) given together tended to increase milk production during the 16 wk of lactation. This effect was more pronounced during the first 4 wk of lactation: 37.5, 37.7, and 40.3 +/- 0.9 kg/d for cows receiving no vitamin supplement, folic acid alone, or folic acid combined with vitamin B(12), respectively. Milk fat yield was not affected by treatments. Lactose, crude protein, and total solid yields were greater, in early lactation, in cows injected with folic acid and vitamin B(12) together but this effect diminished as lactation progressed. Intramuscular injections of folic acid alone or combined with vitamin B(12) tended to decrease plasma concentrations of homocysteine from 5.51 microM with no vitamin supplement to 4.54 and 4.77 +/- 0.37 microM, respectively. Results of the present experiment suggest that the effects of the combined supplement of folic acid and vitamin B(12) on lactational performance of dairy cows were not due to an improvement in methyl groups supply, because RPM supplement, a source of preformed methyl groups, did not alter the cow responsiveness to vitamin supplements. PMID- 19307651 TI - Effect of colostrum immunoglobulin concentration on immunity in Majorera goat kids. AB - The aim of the research was to evaluate the effects of immunoglobulin G (IgG) colostrum concentration on goat kid immune status when the total amount of IgG fed was constant. Majorera goat kids (n = 56) were randomly assigned to 1 of 4 groups, and kids received 4 g of IgG per kg of body weight of atomized colostrum at 4 different IgG concentrations: 20 (AC-20), 40 (AC-40), 60 (AC-60), and 80 (AC 80) mg/mL. Blood samples were obtained on d 0, 1, 2, 3, 4, and 5 postpartum. Immunoglobulin G, IgA, and IgM plasma concentrations, apparent efficiency of absorption of IgG, plasma chitotriosidase activity, plasma complement activity, and plasma proteinogram were measured. Plasma IgG and IgM concentrations were highest on d 1 in AC-80 animals, and IgA plasma concentration was lower in AC-20 than in AC-80. The apparent efficiency of absorption was higher in AC-80 (24.4%) than in the other treatment groups (by an average of 13.8%). Chitotriosidase plasma activity on d 5 (1,488 nmol/mL per hour) was higher than on d 0 and 1 (average of 1,183 nmol/mL per hour). There were no effects of colostrum IgG concentration on complement activity and plasma protein distribution, but gamma globulin and alpha-globulin were lower on d 0 than on d 1, 2, 3, 4, and 5. Increasing the immunoglobulin concentration in colostrum using atomized colostrum improves the immunoglobulin absorption at the same amount of immunoglobulin fed. PMID- 19307652 TI - Near-infrared reflectance spectroscopy prediction of neutral detergent-soluble carbohydrates in timothy and alfalfa. AB - Carbohydrates in forage crops can be divided into neutral detergent-insoluble fiber and neutral detergent-soluble carbohydrates (NDSC); the latter includes organic acids (OA), total ethanol:water-soluble carbohydrates (TESC), starch, and neutral detergent-soluble fiber (NDSF). The accurate and efficient estimation of NDSC in forage crops is essential for improving the performance of dairy cattle. In the present study, visible and near-infrared reflectance spectroscopy (NIRS) were applied to evaluate the feasibility of predicting OA, TESC, starch, NDSF, NDSC, and all related constituents used to calculate these 5 carbohydrate fractions in timothy and alfalfa. Forage samples (n = 1,008) of timothy and alfalfa were taken at the first and second harvests at 2 sites in 2007; samples were dried, ground, and then scanned (400 to 2,500 nm) using an NIRSystems 6500 monochromator. A calibration (n = 60) and a validation (n = 15) set of samples were selected for each species and then chemically analyzed. Concentrations of TESC and NDSC in timothy, as well as starch in alfalfa, were successfully predicted, but many other carbohydrate fractions were not predicted accurately when calibrations were performed using single-species sample sets. Both sets of samples were combined to form new calibration (n = 120) and validation (n = 30) sets of alfalfa and timothy samples. Calibration and validation statistics for the combined sets of alfalfa and timothy samples indicated that TESC, starch, and NDSC were predicted successfully, with coefficients of determination of prediction of 0.92, 0.89, and 0.93, and a ratio of prediction to deviation (RPD) of 3.3, 3.1, and 3.6, respectively. The NDSF prediction was classified as moderately successful The NIRS prediction of OA was unsuccessful All related constituents were predicted successfully by NIRS except ethanol-insoluble residual OM, with Our results confirm the feasibility of using NIRS to predict NDSC, its fractions, and other related constituents, except for OA and ethanol insoluble residual OM, in timothy and alfalfa forage samples. PMID- 19307653 TI - Alfalfa pellet-induced subacute ruminal acidosis in dairy cows increases bacterial endotoxin in the rumen without causing inflammation. AB - A study was conducted to determine if subacute ruminal acidosis (SARA) induced by feeding alfalfa pellets results in increases in free bacterial lipopolysaccharide (LPS) in rumen fluid and peripheral blood, and acute phase proteins in plasma, and to determine the effect of alfalfa pellet-induced SARA on feed intake, rumen fermentation characteristics, milk production and composition, and blood metabolites. Eight lactating Holstein cows, 4 of which were ruminally cannulated, were used in a 6-wk experiment and were fed once daily at 0900 h. During wk 1, cows received a diet containing 50% of DM as concentrate and 50% of DM chopped alfalfa hay. Between wk 2 and wk 6, alfalfa hay was gradually replaced with alfalfa pellets at the rate of 8% per week to reduce rumen pH. Rumen pH was monitored continuously in the ruminally cannulated cows using indwelling pH probes. Rumen fluid and peripheral blood were sampled 15 min before feed delivery and at 6 h after feed delivery. Based on adopted threshold of SARA of at least 180 min/d below pH 5.6, SARA was induced from wk 3 onwards. Replacing 40% of alfalfa hay with alfalfa pellets quadratically increased the DMI from 18.1 kg/d in wk 1 to 23.4 kg/d in wk 6. This replacement linearly decreased milk yield (32.7 vs. 35.9 kg/d) and milk fat percentage and yield (2.32 vs. 3.22%, and 0.77 vs. 1.14 kg/d, respectively), but increased milk protein percentage and yield (3.80 vs. 3.04%, and 1.23 vs. 1.07 kg/d, respectively). This gradual replacement also linearly increased the daily averages of total volatile fatty acids (90 to 121.9 mM), acetate (53.9 to 66.8 mM), propionate (21.5 to 39.6 mM), and osmolality (277.7 to 293.8 mmol/kg) in the rumen and decreased the acetate to propionate ratio from 2.62 to 1.73. Replacing alfalfa hay with alfalfa pellets linearly increased blood lactate from 1.00 mM in wk 1 to a peak of 3.46 mM in wk 5. Induction of SARA in this study increased free rumen LPS concentration from 42,122 endotoxin unit (EU)/mL in wk 1 to 145,593 EU/mL in wk 6. However, this increase was not accompanied by an increase in LPS (<0.05 EU/mL) and in acute phase proteins serum amyloid-A, haptoglobin, and LPS-binding protein in peripheral circulation. Results suggest that SARA induced by alfalfa pellets increased LPS in the rumen without causing translocation of LPS and an immune response. PMID- 19307654 TI - Effects of feeding Fermenten on ruminal fermentation in lactating Holstein cows fed two dietary sugar concentrations. AB - This study was conducted to determine the effects of feeding Fermenten (Church and Dwight Co., Princeton, NJ) with or without dietary sucrose on ruminal fermentation, apparent total-tract nutrient digestibility, and nutrient utilization. Eight ruminally cannulated Holstein cows (163 +/- 55 d in milk; mean +/- standard deviation) were used in a replicated 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. Experimental diets were formulated with and without Fermenten (0 vs. 3.3% of dietary DM) at 2 dietary sugar concentrations (2.8 vs. 5.7%). Dietary treatment did not affect dry matter intake or apparent total-tract nutrient digestibility. Feeding Fermenten did not affect ruminal pH, but high-sugar diets tended to increase the daily minimum pH (5.61 vs. 5.42) and mean pH (6.17 vs. 6.30) compared with low-sugar diets. Ruminal ammonia concentration tended to be greater for cows fed Fermenten compared with control (18.1 vs. 15.9 mg/dL), but was not affected by dietary sugar concentration. Significant interactions between Fermenten and dietary sugar concentration were detected for some milk production responses. Fermenten treatment numerically increased milk fat yield (0.92 vs. 0.82 kg/d), 4% fat corrected milk yield (24.3 vs. 21.9 kg/d), and milk energy output (18.2 vs. 16.4 Mcal/d) compared with control for cows fed low-sugar diets, but not for cows fed high-sugar diets. Increasing dietary sugar concentration did not enhance the effects of Fermenten, providing no support for the theory that synchronizing the availability of N and fermentable energy in the rumen improves nutrient utilization in lactating dairy cows. PMID- 19307655 TI - Effect of a mixture of supplemental dietary plant essential oils on performance of periparturient and early lactation dairy cows. AB - Plant essential plant oils (EO) are volatile aromatic compounds with antimicrobial activity that can alter ruminal fermentation when used as dietary supplements. A feeding trial was conducted to determine the effects of dietary supplementation of periparturient and early lactation dairy cows with a specific mixture of EO. Forty multiparous Holstein cows were randomly assigned to either control (C) or EO-supplemented (1.2 g/cow per day) total mixed rations (TMR). Feeding of treatment diets commenced 3 wk before the expected calving date and continued through 15 wk in lactation. The prepartum TMR contained 70% forage [70% corn silage, 15% alfalfa silage, and 15% wheat straw; dry matter (DM) basis]. The lactation TMR contained 50% forage (60% corn silage, 33% alfalfa silage, 7% alfalfa hay; DM basis). Prepartum and lactation TMR were formulated to contain 12 and 17% CP (DM basis), respectively. There were no differences between treatments for prepartum DM intake (DMI), but DMI was 1.8 kg/d less for EO than C on average across the 15-wk lactation trial. Plasma concentrations of glucose, nonesterified fatty acids, beta-hydroxybutyrate, and urea-N on samples collected -21, -14, -7, 1, 1, 8, 15, 22, and 29 d relative to calving were unaffected by treatment. There were no differences between treatments for actual or fat-corrected milk yields on average across the 15-wk lactation trial. Milk protein content was 0.15% units less for EO than C. Feed efficiency (kg of milk per kg of DMI) tended to be greater for EO than C on average and was greater during wk 8 to 14 of lactation. Prepartum and lactation body weight and condition score measurements were unaffected by treatment. There was no benefit to EO in prepartum dairy cows. Dietary supplementation with EO reduced DMI in early lactation dairy cows with no effect on milk yield. PMID- 19307656 TI - The effect of dietary phosphorus on bone development in dairy heifers. AB - Phosphorus requirements, as percent of dietary dry matter for heifers (0.20 0.35%) and endogenous levels of P in feeds (0.20-0.35% of dry matter) are similar, suggesting that supplementation of P in heifer diets may be infrequently required. Because long-term studies are unavailable, 183 Holstein heifers and 182 Holstein x Jersey crossbred heifers were fed diets with (0.39%) and without (0.29%) supplemental P from 4 to 21 mo of age in a replicated pen design. Two subpopulations of heifers were selected mid-trial for intensive measurement of bone development and metabolism. Thirty-two heifers at 628 d (+/-10.0 d) of age, balanced by breed and diet, were evaluated for bone development. External frame measurements included hip height, length, heart girth, hip width, cannon bone circumference, pelvic length, pelvic height, and pelvic width. Tails of heifers were surgically amputated with the 13 and 14th coccygeal vertebrae retained. After tissue removal, the 13th coccygeal vertebrae were scanned using peripheral quantitative computed tomography with cortical, trabecular, and total bone densities determined. A second subpopulation (n = 64) of heifers (375 d +/- 33 d), balanced for breed and diet, were evaluated for serum pyridinoline and osteocalcin to assess systemic bone metabolism. Data were analyzed as a completely randomized design with breed, treatment, and their interaction in the model. External skeletal measurements revealed significant differences in hip height, hip width, heart girth, cannon bone circumference, and pelvic length between Holstein and crossbred heifers. Supplementing P had no effect on external frame measurements, bone density, or bone metabolism markers. Bone P content was lower (18.1 vs. 18.6%) in heifers fed no supplemental P. Data suggest P supplementation to heifers modestly increased bone P content but increased bone P was not reflected in frame growth, bone density, or bone metabolism. PMID- 19307657 TI - Improving passive transfer of immunoglobulins in calves. I: dose effect of feeding a commercial colostrum replacer. AB - The objective of this study was to describe the effects of feeding 1 or 2 doses of a commercially available colostrum-derived colostrum replacer (CR) on passive transfer of immunoglobulins (Ig) in newborn dairy calves, including IgG, IgG1, IgG2, IgA, and IgM. Newborn calves were removed from the dam before suckling and randomly assigned to 1 of 3 treatment groups: group 1 were fed 1 package (100 g of IgG) of CR product, group 2 was fed 2 packages (200 g of IgG) of the same CR product, and group 3 was fed 3.8 L of maternal colostrum. All colostrum treatments were fed using an esophageal tube feeder within 2 h of birth. Blood samples collected before colostrum feeding and at 24 h of age were tested for serum total protein and Ig concentrations. Mean 24-h serum total protein (TP) and IgG concentrations were significantly lower for calves in group 1 (n = 24; TP = 4.9 g/dL, IgG = 9.6 mg/mL) compared with calves in groups 2 or 3. There was no difference in 24-h serum TP or IgG concentrations between calves in group 2 (n = 23; TP = 5.5 g/dL, IgG = 19.0 mg/mL) and calves in group 3 (n = 22; TP = 5.7 g/dL, IgG = 20.7 mg/mL). Fifty-four, 100, and 91% of calves in groups 1, 2 and 3 achieved acceptable passive transfer (24-h serum IgG > or =10 mg/mL), respectively. Statistically significant but numerically small differences existed between calves in groups 2 and 3 for some 24-h serum Ig classes and subclasses (mean serum concentrations of IgG2, IgA, IgM) and for the relative percentages of Ig classes and subclasses (IgA, IgM, and IgG as a percentage of total Ig; IgG1 and IgG2 as a percentage of total IgG). PMID- 19307658 TI - Improving passive transfer of immunoglobulins in calves. II: interaction between feeding method and volume of colostrum fed. AB - The objective of this study was to investigate the effect of method of feeding (nipple bottle vs. esophageal tube feeder) on passive transfer of immunoglobulin (Ig) G when either a large or small volume of colostrum was fed. Newborn bull calves were removed from the dam before suckling and randomly assigned to 1 of 4 colostrum replacer (CR) treatment groups: 1.5 L (100 g of IgG) of CR fed using a nipple bottle (group 1; n = 24); 1.5 L (100 g of IgG) of CR fed using an esophageal tube feeder (group 2; n = 24); 3.0 L (200 g of IgG) of CR fed using a nipple bottle (group 3; n = 24), or 3.0 L (200 g of IgG) of CR fed using an esophageal tube feeder (group 4; n = 25). Blood samples collected at 24 h of age showed that serum IgG levels were significantly greater in calves fed large (3 L) volumes of CR compared with calves fed small (1.5 L) volumes of CR, regardless of feeding method. These differences were attributed to the larger mass of IgG ingested by calves fed 3 L of CR (200 g of IgG) compared with calves fed 1.5 L of CR (100 g of IgG). For calves fed small (1.5 L) volumes of colostrum, serum total protein (TP, g/dL), serum IgG (IgG, mg/mL), acceptable passive transfer rates (APT, %), and apparent efficiency of absorption of IgG (AEA, %) were significantly greater for calves fed with a bottle (TP = 5.3 g/dL; IgG = 12.5 mg/mL; APT = 100%; AEA = 51.1%) compared with calves fed with an esophageal tube feeder (TP = 5.0 g/dL; IgG = 9.8 mg/mL; APT = 41.7%; AEA = 40.5%). However, for calves fed large (3 L) volumes of colostrum, there was no difference in passive transfer indices for calves fed with a bottle (TP = 5.8 g/dL; IgG = 19.7 mg/mL; APT = 100%; AEA = 41.1%) compared with calves fed with an esophageal tube feeder (TP = 5.9 g/dL; IgG = 18.7 mg/mL; APT = 100%; AEA = 39.0%). PMID- 19307659 TI - Technical note: effects of forage protein-binding polyphenols on chemistry of dairy excreta. AB - Forage chemistry can affect intake, digestion, milk production, and manure excretion. Although information is available on the effects of forage protein binding polyphenols on small ruminant production and manure excretion, little information is available for dairy cattle. The objective of this study was to compare fecal and urinary N excretion of diets formulated with alfalfa (Medicago sativa L.) silage versus condensed tannin-containing birdsfoot trefoil (Lotus corniculatus) or o-quinone-containing red clover (Trifolium pratense L.) silages. Significantly higher concentrations of N were excreted in urine by lactating Holstein dairy cows fed red clover and low-tannin birdsfoot trefoil (8.2 g/L) than by cows fed high-tannin birdsfoot trefoil or alfalfa (7.1 g/L). Fecal N concentrations were similar (33.6 g/kg) among all diets. Dairy cows fed red clover had lower rates of urinary N excretion (5.0 g/h) compared with other forages (6.6 g/h). Fecal N excretion rates were lowest for red clover (4.1 g/h), intermediate for alfalfa (5.8 g/h), and greatest for cows fed high- and low tannin birdsfoot trefoil (6.4 g/h). The ratio of fecal N to urinary N was highest for high-tannin trefoil, lowest for alfalfa and red clover, and higher in excreta collected in morning than evening. Concentrations of neutral detergent fiber (NDF) in feces, of N in NDF (NDIN) and acid detergent fiber (ADIN), and relative amounts of NDIN and ADIN excreted in feces were significantly higher from cows fed high-tannin birdsfoot trefoil than the other silage types. Study results imply that collection of excreta for environmental studies needs to consider forage polyphenol and diurnal effects on chemistry of dairy excreta. PMID- 19307660 TI - Genetic parameters for claw and leg health, foot and leg conformation, and locomotion in Danish Holsteins. AB - The purpose of this study was to estimate the genetic correlations among claw and leg health and potential indicator traits. Claw health was defined as absence of heel horn erosion, interdigital dermatitis, interdigital phlegmon, interdigital hyperplasia, laminitis, and sole ulcer. Leg health was defined as absence of hock infection, swollen hock, and bruising. The potential indicators were locomotion and foot and leg conformation, represented by rear leg side view, rear leg rear view, foot angle, and apparent hock quality and bone structure. The study was conducted using records from 429,877 Danish Holstein cows in first lactation. Binary health traits were divided into 3 subcategories: claw health, leg health, and absence of all claw and leg disorders. Genetic (r(g)) and phenotypic correlations were estimated using a bivariate linear sire model and REML. Estimated heritabilities were 0.01 for all 3 combined claw and leg health traits (on the observed binary scale), 0.09 for locomotion, 0.14 for rear leg rear view, 0.19 for rear leg side view, 0.13 for foot angle, 0.22 for apparent hock quality, and 0.27 for apparent bone structure. Heritabilities were 0.06 and 0.01 for claw health and leg health, respectively, when transformed to the underlying continuous scale. Claw and leg disorders are an increasing problem for Danish Holsteins, but genetic improvement of claw and leg health is challenging because the traits have low heritabilities. Claw and leg health were separate but correlated traits (r(g) = 0.35). Locomotion and rear leg rear view were useful indicator traits for claw health (r(g) = 0.46 and r(g) = 0.21, respectively), whereas hock quality and bone structure were useful indicators for leg health (r(g) = 0.42 and 0.26, respectively). Claw and leg health should be considered as separate traits in genetic evaluations that also include the useful indicator traits to compensate for low heritability of the health traits. PMID- 19307661 TI - Inferring relationships between health and fertility in Norwegian Red cows using recursive models. AB - Health and fertility are complex traits, and the phenotype for one trait may affect the phenotype of one or more other traits. For instance, disease in early lactation may impair a cow's ability to show estrus and to conceive after insemination. The objectives of the present study were to explore phenotypic and genetic relationships among health and fertility traits in Norwegian Red cows using a recursive effects model, which allows disentangling causal effects of phenotypes from the genetic and environmental correlations among traits. Records of interval from calving to first insemination (CFI), nonreturn rate within 56 d after first insemination (NR56), clinical mastitis (CM), ketosis (KET), and retained placenta from 55,568 first-lactation daughters of 1,577 Norwegian Red sires were analyzed. Trivariate recursive Gaussian-threshold models were used to analyze the 2 fertility traits (CFI and NR56) together with 1 disease trait in each analysis. The estimated structural coefficients of the recursive models imply that presence of KET or retained placenta lengthened CFI, whereas causal effects from CM to fertility were negligible. Recursive effects of disease on NR56, and of CFI on NR56, were all close to zero. Genetic correlations between health and fertility traits were low or moderate. The strongest genetic correlation was between KET and CFI (0.29), whereas genetic correlations between CM and NR56 and between CFI and NR56 were nil. In general, selection against disease is expected to slightly improve fertility (shorter CFI and higher NR56) as a correlated response and vice versa. The present results suggest that the use of structural-equation models, such as the one used here, may enhance our understanding of complex relationships among traits. PMID- 19307662 TI - Phenotypic and genetic relationships of common health disorders with milk and fat yield persistencies from producer-recorded health data and test-day yields. AB - The objective of this study was to investigate phenotypic and genetic relationships of common health disorders in dairy cows with milk (PMY) and fat (PFY) yield persistencies. Health and production data from 398 commercial dairy herds were used. Disease traits were defined in binary form for individual lactations considering mastitis only during the first 100 d in milk (MAST1), only after 100 d in milk (MAST2), and at any stage of lactation (MAST), and reproductive disorders (REPRO), metabolic disorders (METAB), and lameness (LAME). The persistencies were defined to be uncorrelated with 305-d yields. Impact of the diseases on PMY and PFY were investigated separately in first (FL) and later (LL) lactations. Phenotypic associations of PMY and PFY with likelihood of diseases in current and subsequent lactations were examined using odds ratios from a logistic regression model. Linear-threshold sire-maternal grandsire models were used to estimate genetic correlations of displaced abomasums (DA), ketosis (KET), metritis (MET), MAST, MAST1, and MAST2 with PMY and PFY across parities. Metabolic diseases and REPRO had significantly positive relationships with PMY and PFY in both FL and LL. Significantly greater PMY and PFY were associated with MAST1 in LL. Significantly lower PMY and PFY were related to MAST2 in both FL and LL, whereas cows affected by MAST had significantly less persistent lactations. Incidence of MAST and MAST2 decreased with increasing PMY and PFY in the present and previous lactation. Heritability of disease incidences were 0.03 (DA), 0.01 (KET), 0.10 (MAST), 0.02 to 0.05 (MAST1), 0.02 (MAST2), and 0.04 to 0.10 (MET). Displaced abomasum, KET, MAST, MAST1, and MET had unfavorable genetic correlations of 0.35, 0.46, 0.17, 0.02, and 0.27 with PMY, and 0.16, 0.21, 0.07, 0.06, and 0.12 with PFY, respectively. Favorable genetic correlations were found for MAST2 with PMY (-0.24) and PFY (-0.04). Results suggest that diseases in early lactation increase persistency of milk and fat yield. Selection for greater lactation persistency must consider these antagonistic relationships. PMID- 19307663 TI - Best prediction of yields for long lactations. AB - Lactation records of any reasonable length now can be processed with the selection index method known as best prediction (BP). Previous prediction programs were limited to the 305-d standard used since 1935. Best prediction was implemented in 1998 to calculate lactation records in USDA genetic evaluations, replacing the test interval method used since 1969 to calculate lactation records. Best prediction is more complex but also more accurate, particularly when testing is less frequent. Programs were reorganized to output better graphics, give users simpler access to options, and provide additional output, such as BP of daily yields. Test-day data for 6 breeds were extracted from the national dairy database, and lactation lengths were required to be > or =500 d (Ayrshire, Milking Shorthorn) or > or =800 d (all others). Average yield and SD at any day in milk (DIM) were estimated by fitting 3-parameter Wood's curves (milk, fat, protein) and 4-parameter exponential functions (somatic cell score) to means and SD of 15- (< or =300 DIM) and 30-d (>300 DIM) intervals. Correlations among TD yields were estimated using an autoregressive matrix to account for biological changes and an identity matrix to model daily measurement error. Autoregressive parameters (r) were estimated separately for first (r = 0.998) and later parities (r = 0.995). These r values were slightly larger than previous estimates due to the inclusion of the identity matrix. Correlations between traits were modified so that correlations between somatic cell score and other traits may be nonzero. The new lactation curves and correlation functions were validated by extracting TD data from the national database, estimating 305-d yields using the original and new programs, and correlating those results. Daily BP of yield were validated using daily milk weights from on-farm meters in university research herds. Correlations ranged from 0.900 to 0.988 for 305-d milk yield. High correlations ranged from 0.844 to 0.988 for daily yields, although correlations were as low as 0.015 on d 1 of lactation, which may be due to calving-related disorders that are not accounted for by BP. Correlations between 305-d yield calculated using 50-d intervals from 50 to 250 DIM and 305-yield calculated using all TD to 500 DIM increased as TD data accumulated. Many cows can profitably produce for >305 DIM, and the revised program provides a flexible tool to model these records. PMID- 19307664 TI - Dynamics of endemic infectious diseases of animal and human importance on three dairy herds in the northeastern United States. AB - Endemic infectious diseases in dairy cattle are of significant concern to the industry as well as for public health because of their potential impact on animal and human health, milk and meat production, food safety, and economics. We sought to provide insight into the dynamics of important endemic infectious diseases in 3 northeastern US dairy herds. Fecal samples from individual cows and various environmental samples from these farms were tested for the presence of major zoonotic pathogens (i.e., Salmonella, Campylobacter, and Listeria) as well as commensal bacteria Escherichia coli and enterococci. Additionally, the presence of Mycobacterium avium ssp. paratuberculosis was tested in fecal and serum samples from individual cows. Test results and health and reproductive records were maintained in a database, and fecal, plasma, DNA, and tissue samples were kept in a biobank. All bacteria of interest were detected on these farms and their presence was variable both within and between farms. The prevalence of Listeria spp. and L. monocytogenes in individual fecal samples within farm A ranged from 0 to 68.2% and 0 to 25.5%, respectively, over a period of 3 yr. Within farm B, continuous fecal shedding of Salmonella spp. was observed with a prevalence ranging from 8 to 88%; Salmonella Cerro was the predominant serotype. Farm C appeared less contaminated with Salmonella and Listeria, although in the summer of 2005, 50 and 19.2% of fecal samples were positive for Listeria and L. monocytogenes, respectively. The high prevalence of E. coli (89 to 100%), Enterococcus (75 to 100%), and Campylobacter (0 to 81%) in feces suggested they were ubiquitous throughout the farm environment. Fecal culture and ELISA results indicated a low prevalence of Mycobacterium avium ssp. paratuberculosis infection in these farms (0 to 13.6% and 0 to 4.9% for culture-positive and ELISA-positive, respectively), although the occasional presence of high shedders was observed. Results have major implications for food safety and epidemiology by providing a better understanding of infectious disease dynamics on dairy farms. Comprehensive understanding of these infections may lead to better farm management practices and pathogen reduction programs to control and reduce the on-farm contamination of these pathogens and to prevent their further entry into the food-chain. PMID- 19307667 TI - Comparable survival between HIV+ and HIV- non-Hodgkin and Hodgkin lymphoma patients undergoing autologous peripheral blood stem cell transplantation. AB - Autologous stem cell transplantation (ASCT) has been successfully used in HIV related lymphoma (HIV-Ly) patients on highly active antiretroviral therapy. We report the first comparative analysis between HIV-Ly and a matched cohort of HIV( ) lymphoma patients. This retrospective European Group for Blood and Marrow Transplantation study included 53 patients (66% non-Hodgkin and 34% Hodgkin lymphoma) within each cohort. Both groups were comparable except for the higher proportion of males, mixed-cellularity Hodgkin lymphoma and patients receiving granulocyte colony-stimulating factor before engraftment and a smaller proportion receiving total body irradiation-based conditioning within the HIV-Ly cohort. Incidence of relapse, overall survival, and progression-free survival were similar in both cohorts. A higher nonrelapse mortality within the first year after ASCT was observed in the HIV-Ly group (8% vs 2%), predominantly because of early bacterial infections, although this was not statistically significant and did not influence survival. Thus, within the highly active antiretroviral therapy era, HIV patients should be considered for ASCT according to the same criteria adopted for HIV(-) lymphoma patients. PMID- 19307668 TI - The BCL6 transcriptional program features repression of multiple oncogenes in primary B cells and is deregulated in DLBCL. AB - The BCL6 transcriptional repressor is required for development of germinal center (GC) B cells and when expressed constitutively causes diffuse large B-cell lymphomas (DLBCLs). We examined genome-wide BCL6 promoter binding in GC B cells versus DLBCLs to better understand its function in these settings. BCL6 bound to both distinct and common sets of functionally related gene in normal GC cells versus DLBCL cells. Certain BCL6 target genes were preferentially repressed in GC B cells, but not DLBCL cells. Several such genes have prominent oncogenic functions, such as BCL2, MYC, BMI1, EIF4E, JUNB, and CCND1. BCL6 and BCL2 expression was negatively correlated in primary DLBCLs except in the presence of BCL2 translocations. The specific BCL6 inhibitor retro-inverso BCL6 peptidomimetic inhibitor-induced expression of BCL2 and other oncogenes, consistent with direct repression effects by BCL6. These data are consistent with a model whereby BCL6 can directly silence oncogenes in GC B cells and counterbalance its own tumorigenic potential. Finally, a BCL6 consensus sequence and binding sites for other physiologically relevant transcription factors were highly enriched among target genes and distributed in a pathway-dependent manner, suggesting that BCL6 forms specific regulatory circuits with other B-cell transcriptional factors. PMID- 19307669 TI - Controversies in the treatment of knee dislocations and multiligament reconstruction. AB - A systematic approach to evaluation and treatment is needed for the patient with knee dislocation. There is a paucity of high-level evidence on which to base treatment decisions. Reported controversies related to the treatment of the multiligament-injured knee include the selective use of arteriography for vascular assessment, serial physical examination with the ankle-brachial index, acute surgical treatment of all damaged structures, the selective application of preoperative and postoperative joint-spanning external fixation, arthroscopic reconstruction of the anterior cruciate ligament and posterior cruciate ligament, simultaneous open reconstruction with repair of the posterolateral corner, reconstruction and/or repair of the posteromedial corner, and the use of allograft tissue. PMID- 19307670 TI - Acromioclavicular joint injuries: diagnosis and management. AB - Acromioclavicular joint injuries represent nearly half of all athletic shoulder injuries, often resulting from a fall onto the tip of the shoulder with the arm in adduction. Stability of this joint depends on the integrity of the acromioclavicular ligaments and capsule as well as the coracoclavicular ligaments and the trapezius and deltoid muscles. Along with clinical examination for tenderness and instability, radiographic examination is critical in the evaluation of acromioclavicular joint injuries. Nonsurgical treatment is indicated for type I and II injuries; surgery is almost always recommended for type IV, V, and VI injuries. Management of type III injuries remains controversial, with nonsurgical treatment favored in most instances and reconstruction of the acromioclavicular joint reserved for symptomatic instability. Recommended techniques for stabilization in cases of acute and late symptomatic instability include screw fixation of the coracoid process to the clavicle, coracoacromial ligament transfer, and coracoclavicular ligament reconstruction. Biomechanical studies have demonstrated that anatomic acromioclavicular joint reconstruction is the most effective treatment for persistent instability. PMID- 19307671 TI - Distal fibula malunions. AB - Anatomic reduction and fixation of unstable ankle fractures is necessary to prevent posttraumatic arthritis. Malunion of the distal fibula in unstable ankle fractures may lead to progressive talar instability. Ankle fracture malunions often present with concomitant syndesmotic widening, which can cause surgeons to overlook changes in fibula length and rotation. The decision to proceed with surgery should be made only after a careful diagnostic workup and detailed preoperative discussion with the patient. Considerations for surgical management include location and orientation of a corrective osteotomy, use of structural graft, widening of the syndesmosis, assessment of reduction, and the need for medial exposure. Good and excellent clinical results after fibular reconstruction have been reported in 67% to 92% of ankles. Proper patient selection is critical, because ankle malunions can be complicated, with coexisting fibular, syndesmotic, medial, and posterior malleolar malalignment, along with degenerative joint disease. Understanding the indications and surgical technique for revising fibular malunions may obviate a future salvage procedure. PMID- 19307672 TI - Achondroplasia: manifestations and treatment. AB - Achondroplasia, the most common skeletal dysplasia, is caused by a mutation of fibroblast growth factor receptor-3. This disorder is characterized by frontal bossing, midface hypoplasia, otolaryngeal system dysfunction, and rhizomelic short stature. Orthopaedic manifestations are exhibited in the spine and the extremities. In the infant with achondroplasia, foramen magnum stenosis may result in brainstem compression with apnea and sudden death. Thoracolumbar kyphosis is seen in most infants, but typically it resolves when the child begins to walk. Anatomic anomalies of the vertebral column place the patient at risk for spinal stenosis as early as the first decade and especially during adulthood. Radial head dislocation is one manifestation in the upper extremity. Lower extremity alignment often is characterized by genu varum, which may require correction osteotomy. Medical and surgical options are available to increase patient height, but indications are controversial, and treatment often consumes a large portion of the child's life. PMID- 19307673 TI - Shoulder problems in children with brachial plexus birth palsy: evaluation and management. AB - Traction injury to the brachial plexus sustained during the birth process that results in impaired neuromuscular function of the upper extremity continues to occur despite advances in modern obstetric care. The most common pattern of injury usually results in motor weakness of shoulder external rotation, leading to internal rotation contractures and subsequent deformity of the skeletally immature glenohumeral joint. Understanding of these deformities and effective surgical intervention have advanced greatly over the past decade. Restoration of balance between internal and external rotation forces around the shoulder has great potential for remodeling of the glenohumeral joint in the young child. Arthroscopic-directed release of the contracture, with select use of latissimus dorsi transfer to provide external rotation power, has proved to be effective for many children with these contractures. PMID- 19307674 TI - Pathoanatomy and clinical correlates of the immunoinflammatory response following orthopaedic trauma. AB - The natural inflammatory response to major trauma may be associated with the development of a systemic inflammatory state, remote multiorgan failure, and death. Although a controlled inflammatory response is beneficial, an exaggerated response can cause serious adverse systemic effects. Early identification of high risk patients, based on inflammatory markers and genomic predisposition, should help direct intervention in terms of surgical stabilization and biologic response modification. Currently, two markers of immune reactivity, interleukin-6 and human leukocyte antigen-DR class II molecules, appear to have the most potential for regular use in predicting the clinical course and outcome in trauma patients; however, the ability to measure markers of inflammation is still limited at many hospitals. With improving technology and increasing research interest, understanding of the significance of the immunoinflammatory response system in injured patients will continue to evolve. PMID- 19307676 TI - Posterior reversible encephalopathy syndrome associated with methotrexate neurotoxicity: conventional magnetic resonance and diffusion-weighted imaging findings. AB - The addition of intrathecal methotrexate to treatment protocols has increased survival rates in children with acute lymphoblastic leukemia but is also associated with varying degrees of neurotoxicity. We describe a 15-year-old female patient diagnosed with acute lymphoblastic leukemia presenting with status epilepticus after receiving intrathecal methotrexate. Magnetic resonance imaging showed reversible cortical and subcortical changes consisting of high-intensity lesions on T2-weighted and fluid-attenuated inversion recovery sequences with postgadolinium enhancement, low signal intensity on diffusion-weighted imaging and increased apparent diffusion coefficient. These findings were consistent with the posterior reversible encephalopathy syndrome. We report our conventional magnetic resonance and diffusion-weighted imaging findings and briefly discuss the pathophysiology of the syndrome. PMID- 19307677 TI - Is there still a place for pharmacological testing for phaeochromocytoma? PMID- 19307678 TI - Systemic treatment in hepatocellular carcinoma; 'A small step for man...'. AB - Hepatocellular carcinoma (HCC) is the third most common cause of cancer mortality worldwide. In localised disease, orthotopic liver transplantation, surgical resection or local ablations are the mainstay of treatment. In unresectable or metastatic HCC, systemic therapy has unfortunately yielded disappointing results and therefore until recently was generally considered to be ineffective. Most patients with HCC have an underlying liver disease and many drugs may exacerbate the underlying liver disease. Recently, two randomised phase II trials with sorafenib in patients with advanced or metastatic HCC have shown a significant increase in progression free and overall survival of approximately two months, which is an absolute novum for this disease. Sorafenib is therefore now considered a viable treatment option in patients with unresectable or metastatic HCC, a good performance status and Child-Pugh A liver cirrhosis. Despite this very promising result, of major concern is the treatment-related toxicity as observed in these and other trials by sorafenib treatment. However, the important first significant survival benefit by systemic treatment has generated hope for the development of new treatment strategies which will be more efficacious, have favourable toxicity profiles and will further extend survival of this still highly lethal disease. PMID- 19307679 TI - Evaluation of Endocrine Tests. C: glucagon and clonidine test in phaeochromocytoma. AB - BACKGROUND: The diagnosis of phaeochromocytoma is based on the demonstration of catecholamine excess. Urine and plasma metanephrine measurements are highly sensitive tests for the diagnosis of phaeochromocytoma, but moderate elevations in metanephrines lack optimal specificity. In this study we aimed to evaluate the diagnostic value of additional tests, i.e. glucagon stimulation and clonidine suppression test, in patients with moderately elevated catecholamines and/or metanephrines. METHODS: Patients with suspected phaeochromocytoma with moderately elevated catecholamines and/or metanephrines in plasma or urine were subjected to the glucagon stimulation and clonidine suppression test. The presence of phaeochromocytoma was confirmed by histology and the absence by a disease-free extended follow-up. RESULTS: Fifty-five patients were included. Phaeochromocytoma was diagnosed in 11 patients. The follow-up period in patients without phaeochromocytoma was 56 (19 to 154) months. The sensitivity of the glucagon test was 30% and the specificity 100%. The clonidine test had no discriminative power, because the area under the ROC curve was not significantly different from 0.5. CONCLUSION: The clonidine suppression test without normetanephrine measurements and the glucagon stimulation test are not sensitive enough to safely exclude phaeochromocytoma in patients with mildly elevated plasma or urine catecholamines. PMID- 19307680 TI - Seven-day PPI-triple therapy with levofloxacin is very effective for Helicobacter pylori eradication. AB - BACKGROUND: Helicobacter pylori infection causes lifelong gastritis and is associated with the development of peptic ulcer disease, MALT lymphoma and gastric cancer. Many patients benefit from H. pylori eradication therapy. PPI triple therapy is recommended as initial therapy. Quadruple therapy, with a PPI, bismuth, and two antibiotics, used to be recommended as second-line therapy, but can no longer be prescribed because bismuth is no longer available. Therefore, there is an urgent need for new effective rescue therapies. Levofloxacin-based therapies were suggested as an alternative to quadruple therapy. The aim of this study is to examine the efficacy and tolerability of such a one-week therapy with levofloxacin and esomeprazole combined with either amoxicillin or clarithromycin in a Dutch population. METHODS: Between February 2005 and November 2006, 123 consecutive H. pylori positive patients were enrolled in this study. The first 59 patients were treated with esomeprazole, amoxicillin and levofloxacin (group I). The next 64 patients were treated with esomeprazole, clarithromycin, and levofloxacin (group II ). Both therapies were compared for efficacy and tolerability. RESULTS: In group I the overall (ITT) cure rate was 96% and in group II it was 93%. Minor side effects occurred in 29% of patients in group I and in 41% of patients in group II. Major side effects that warranted discontinuation of therapy occurred in two patients in group II. CONCLUSION: Seven-day triple therapy with esomeprazole, levofloxacin and either amoxicillin or clarithromycin for seven days is very effective and safe for H. pylori eradication. The combination with amoxicillin seems to be better tolerated than the combination with clarithromycin. PMID- 19307681 TI - Symptomatic hypoparathyroidism based on a 22q11 deletion first diagnosed in a 43 year-old woman. AB - Congenital hypoparathyroidism usually manifests in early childhood with hypocalcaemia with or without clinical characteristics. This report describes a Caucasian woman who, at the age of 43 years, was diagnosed with dysgenesis of the parathyroid glands due to a de novo microdeletion in chromosome 22q11 or DiGeorge syndrome. This syndrome is characterised by a considerable variability in clinical symptoms, including heart defects, thymic hypoplasia and mental retardation. Our patient presented with generalised convulsions due to extreme, symptomatic hypocalcaemia. The convulsions had been apparent for 18 months at the time of the diagnosis. Remarkably, whereas parathyroid hormone levels were undetectable, the 1,25-dihydroxy vitamin D level was normal. Chromosome 22q11 deletion was confirmed by fluorescence in situ hybridisation analysis. PMID- 19307682 TI - Hypocalcaemia as presenting symptom of velocardiofacial syndrome. AB - Hypocalcaemia due to hypoparathyroidism is a rare finding in adults. The coexistence of cardiac abnormalities may be suggestive of a hereditary syndrome. We describe a case of velocardiofacial syndrome in a woman without a family history of this disorder. The hypocalcaemia was treated with calcium and vitamin D supplementation. PMID- 19307683 TI - Anti-TNFalpha and agranulomatous tuberculous manifestations: more diagnostic dilemma. PMID- 19307684 TI - Whipple's disease: easily diagnosed, if considered. PMID- 19307685 TI - Autoimmune haemolytic anaemia due to chronic hepatitis C virus infection treated with prednisone, pegylated interferon and ribavirin. PMID- 19307686 TI - A woman with a painful and swollen hand. PMID- 19307687 TI - Ceftriaxone-associated biliary pseudolithiasis. PMID- 19307688 TI - The past, the present and the future of experimental research on myocardial ischemia and protection. AB - At present, cardiovascular diseases represent the most important health risks because they are responsible for more than 50% of total mortality. Among them, ischemic heart disease is the leading cause of morbidity and mortality, and according to the World Health Organization, will be the major global cause of death by the year 2020. Major progress in the prognosis, diagnosis and therapy of ischemic heart disease would be impossible without notable achievements of the 20th century that have been critical for further development of cardiology.We are now living in the era of molecular medicine, and the influence of basic research on clinical practice has never been more pronounced. This, however, necessitates a new strategy; future cardiovascular research should include the following general guidelines: 1) to evaluate the role and proportion of already described molecular pathways; descriptive approaches will gradually disappear; 2) to distinguish between acute, chronic and pleiotropic effects of different drugs under in vitro and in vivo conditions, with respect to possible clinical use; 3) to use clinically relevant genetic models; 4) to study possible alterations in intracellular signaling in order to find the decisive steps responsible for abnormal control of cell growth, contractile function, lipid metabolism, cardiac ischemic tolerance, etc.; 5) to study the molecular mechanisms of cardiovascular diseases not only in healthy individuals, but also under different pathological conditions. Such an approach must include developmental and gender differences, which are particularly important for the field of ischemic heart disease; therefore, experimental cardiovascular research can no longer be restricted to males of uncertain age. It is hoped that patients in future decades will profit from the progress of basic cardiovascular research. PMID- 19307689 TI - Effect of blockage of the endocannabinoid system by CB(1) antagonism on cardiovascular risk. AB - The endocannabinoid system is a crucial player in the inflammatory processes underlying atherosclerosis. Recently, basic research studies and animal models have strongly supported the role of the endocannabinoid system not only in the regulation of classical cardiovascular risk factors (including lipid profile and glucose homeostasis), but also in the activation of immune cells and inflammatory mediators. Clinical trials investigating treatment with rimonabant (a selective antagonist of the cannabinoid type 1 receptor) have suggested a beneficial effect of this drug in the management of obesity. Further studies are needed to explore a possible use for rimonabant in treating type 2 diabetes and acute and chronic cardiovascular disease. Despite the slight increase in adverse events (mainly psychiatric), which has led to the recent withdrawal of rimonabant from the market, CB(1) receptor antagonism might represent a very promising therapeutic strategy to reduce the cardiovascular risk. In the present review, we focused on the most important experimental investigations into the role of the endocannabinoid system in atherosclerosis and cardiovascular risk. PMID- 19307690 TI - ICAM-1 signaling in endothelial cells. AB - Intercellular adhesion molecule-1 (ICAM-1; CD54) is a 90 kDa member of the immunoglobulin (Ig) superfamily and is critical for the firm arrest and transmigration of leukocytes out of blood vessels and into tissues. ICAM-1 is constitutively present on endothelial cells, but its expression is increased by proinflammatory cytokines. The endothelial expression of ICAM-1 is increased in atherosclerotic and transplant-associated atherosclerotic tissue and in animal models of atherosclerosis. Additionally, ICAM-1 has been implicated in the progression of autoimmune diseases. We and others have shown that the ligation of ICAM-1 on the surface of endothelial or smooth muscle cells with monoclonal antibodies, via its main leukocyte ligand, lymphocyte function associated molecule (LFA)-1, or with antibodies derived from patient serum, leads to the activation of several proinflammatory signaling cascades, and to the rearrangement of the actin cytoskeleton. A circulating or soluble form of ICAM-1 (sICAM-1) has been measured in various body fluids, with elevated levels being observed in patients with atherosclerosis, heart failure, coronary artery disease and transplant vasculopathy. sICAM-1 has signaling properties in several cell types, including EC, and invokes a range of proinflammatory responses. Thus, we propose that in addition to acting as a leukocyte adhesion molecule, ICAM-1 directly contributes to inflammatory responses within the blood vessel wall by increasing endothelial cell activation and augmenting atherosclerotic plaque formation. PMID- 19307691 TI - Nitrate tolerance as a model of vascular dysfunction: roles for mitochondrial aldehyde dehydrogenase and mitochondrial oxidative stress. AB - Organic nitrates are a group of very effective anti-ischemic drugs. They are used for the treatment of patients with stable angina, acute myocardial infarction and chronic congestive heart failure. A major therapeutic limitation inherent to organic nitrates is the development of tolerance, which occurs during chronic treatment with these agents. The mechanisms underlying nitrate tolerance remain incompletely defined and are likely multifactorial. One mechanism seems to be a diminished bioconversion of nitroglycerin, another seems to be the induction of vascular oxidative stress, and a third may include neurohumoral adaptations. Recent studies have revealed that mitochondrial reactive oxygen species (ROS) formation and a subsequent oxidative inactivation of nitrate reductase, the mitochondrial aldehyde dehydrogenase (ALDH-2), play an important role in the development of nitrate and cross-tolerance. The present review focus first on the role of oxidative stress and second on the role of ALDH-2 in organic nitrate bioactivation leading to the development of tolerance and cross-tolerance (endothelial dysfunction) in response to nitroglycerin treatment. Recently, the role of mitochondrial oxidative stress in the development of nitrate tolerance was demonstrated in a mouse model with a heterozygous deletion of manganese superoxide dismutase (MnSOD(+/-)), which is the mitochondrial isoform of this enzyme. Studies from our own laboratory have provided evidence for cross-talk between mitochondrial and cytosolic (Nox-dependent) sources of ROS. We close this review by focusing on the protective properties of the organic nitrate pentaerithrityl tetranitrate, which upregulates enzymes that have strong antioxidative activity, such as heme oxygenase-1 and ferritin, thereby preventing the development of tolerance and endothelial dysfunction. PMID- 19307692 TI - Therapeutic potential of plasma membrane-derived microparticles. AB - In the past, plasma membrane-derived microparticles were considered "cellular dust." According to the literature, circulating levels of microparticles are increased in several cardiovascular diseases associated with inflammation, suggesting that microparticles are linked to deleterious effects such as endothelial dysfunction or thrombosis. However, very recent studies have shown that under several conditions microparticles can transfer biological messages between cells. Indeed, microparticles act as vectors of key information to maintain cell homeostasis or to favor cell repair and induce angiogenesis. For instance, microparticles of platelet origin are able to repair myocardial injury after myocardial infarction. Also, we have shown that engineered microparticles generated from human activated/apoptotic T cells promote angiogenesis through the up-regulation of adhesion proteins and pro-angiogenic factors in human endothelial cells. Interestingly, the effects induced by these microparticles on the formation of capillary-like structures, expression of adhesion molecules, and pro-angiogenic factors are reversed after silencing of the Sonic Hedgehog (Shh) morphogen pathway. In addition, the same type of microparticles is able to induce neo-vascularization in an ischemic hindlimb model. These effects are, at least in part, mediated by Shh and nitric oxide production. Taking into consideration these results and the most recent data concerning the ability of microparticles to transmit genetic information between cells through mRNA transfer, it is plausible that plasma membrane-derived microparticles could serve as tools with veritable therapeutic potential. PMID- 19307693 TI - Effects of aldosterone on coronary function. AB - Our understanding of the effects of aldosterone and its mechanisms has increased substantially in recent years, probably because of the importance of the mineralocorticoid receptor (MR) antagonists in several major cardiovascular diseases. Recent clinical studies have confirmed the benefits of MR antagonists in patients with heart failure, left ventricular dysfunction after myocardial infarction, hypertension or diabetic nephropathy. However, it would be a gross oversimplification to conclude that the role of aldosterone is unequivocally negative. Aldosterone is synthesized in the adrenal glands and binds to specific MRs in target epithelial cells. The steroid-receptor complex penetrates the cell nucleus where it modulates gene expression and activates specific aldosterone induced proteins that control sodium reabsorption. Recent studies have shown that aldosterone also impacts a wide range of non-epithelial tissues such as the heart and blood vessels. Remarkably, aldosterone can also be synthesized in extra adrenal tissues and it may act in a rapid non-genomic manner.We note the existence of glucocorticoids that exhibit plasma concentrations much higher than those of aldosterone and that are structurally very similar to aldosterone. It is thus possible that glucocorticoids may bind to the aldosterone receptor in some cell types. Diverse experimental models and several strains of transgenic mice have allowed us to better understand the effects of aldosterone on the heart. Specifically, it seems that a slight increase in cardiac aldosterone concentrations induces a decreased coronary reserve in mice by decreasing the BKCa potassium channels associated with coronary smooth muscle cells. Taken together, these experiments indicate that vascular cells are the primary targets of aldosterone in the cardiovascular system. The hormone directly affects NO and EDHF-mediated coronary relaxation. Both mechanisms may contribute to the deleterious cardiovascular effects of MR stimulation. PMID- 19307694 TI - Antihypertensive effects of the flavonoid quercetin. AB - The blood pressure lowering effect of a fruit and vegetable-rich diet is a necessary dietary lifestyle measure now included the guidelines for the management of arterial hypertension. Furthermore, flavonoids represent a major class of plant polyphenolics. The present review addresses the antihypertensive effect of quercetin, one of the most abundant flavonoids present in fruits and vegetables, and probably the best studied flavonoid because of its high biological activity. Quercetin has been shown to induce a progressive, dose dependent and sustained reduction in blood pressure when given chronically in several rat models of hypertension, including spontaneously hypertensive rats, L NAME-treated rats, DOCA-salt hypertensive rats, two-kidney one-clip Goldblatt rats, rats with aortic constriction and Dahl salt-sensitive hypertensive rats. Quercetin was also effective in reducing blood pressure in rat models of metabolic syndrome, including the obese Zucker rats as well as rats treated with a high-sucrose, high-fat diet. Quercetin also prevented morphological and functional changes in the heart, vessels and kidney, while increasing production of reactive oxygen species associated with hypertension. A high dose of quercetin also reduced blood pressure in stage 1 hypertensive patients in a randomized, double-blind, placebo-controlled, crossover study. Since raised blood pressure is the major cause of stroke as well as an important risk factor for ischemic heart disease, we propose that the blood pressure-lowering effect of quercetin could be an important mechanism contributing to the reduced risk of myocardial infarction and stroke observed with fruit and vegetables-rich diets, and possibly with flavonoid-rich diets. PMID- 19307695 TI - Activation of nicotinamide N-methyltrasferase and increased formation of 1 methylnicotinamide (MNA) in atherosclerosis. AB - Nicotinamide N-methyltrasferase (NMMT) catalyzes the conversion of nicotinamide (NA) to 1-methylnicotinamide (MNA). Recent studies have reported that exogenous MNA exerts anti-thrombotic and anti-inflammatory activity, suggesting that endogenous NMMT-derived MNA may play a biological role in the cardiovascular system. In the present study, we assayed changes in hepatic NNMT activity and MNA plasma levels along the progression of atherosclerosis in apoE/LDLR(-/-) mice, as compared to age-matched wild-type mice. Atherosclerosis progression in apoE/LDLR( /-) mice was quantified in aortic root, while hepatic NNMT activity and MNA plasma concentrations were concomitantly measured in 2-, 3-, 4-, and 6-month-old mice. In apoE/LDLR(-/-) mice, atherosclerotic plaques developed in the aortic roots beginning at the age of 3 months and gradually increased in size, macrophage content, and inflammation intensity over time, as detected by Oil-Red O staining, CD68 immunostaining, and in situ zymography (MMP2/MMP9 activity). Hepatic NNMT activity was upregulated approximately two-fold in apoE/LDLR(-/-) mice by the age of 2 months, as compared to wild-type mice (1.03 +/- 0.14 vs. 0.64 +/- 0.23 pmol/min/mg, respectively). MNA plasma concentrations were also elevated approximately two-fold (0.30 +/- 0.13 vs. 0.17 +/- 0.04 micromol/l, respectively). As atherosclerosis progressed, hepatic NMMTactivity and MNA plasma concentrations increased five-fold in 6-month-old apoE/LDLR(-/-) mice at the stage of advanced atherosclerotic plaques (NMMT activity: 2.29 +/- 0.34 pmol/min/mg, MNA concentration: 1.083 +/- 0.33 micromol/l). In summary, the present study demonstrated that the progression of vascular inflammation and atherosclerosis was associated with the upregulation of hepatic NNMT activity and subsequent increase in endogenous MNA plasma levels. Given the anti-thrombotic and anti-inflammatory properties of exogenous MNA, robust activation of an endogenous NA-MNA pathway in atherosclerosis may play an important compensatory role. PMID- 19307696 TI - Anti-diabetic effects of 1-methylnicotinamide (MNA) in streptozocin-induced diabetes in rats. AB - 1-Methylnicotinamide (MNA), a major endogenous metabolite of nicotinamide, possesses anti-thrombotic and anti-inflammatory activity, and reverses endothelial dysfunction. In the present work, we investigated whether such a vasoprotective profile of MNA activity affords anti-diabetic action in rats. Diabetes was induced by streptozotocin (STZ) in Sprague-Dawley rats. Eight weeks after STZ injection in untreated or MNA-treated rats (100 mg kg(-1) daily), development of diabetes (plasma concentrations of fasting and non-fasting glucose, HbA(1c), peptide C), development of oxidant stress (lipid peroxidation, carbonylation of plasma proteins), as well as NO-dependent endothelial function in aorta, coronary and mesenteric vessels were analyzed. Finally, the effect of chronic treatment with MNA on long-term survival of diabetic rats was determined. Chronic treatment with MNA profoundly lowered fasting glucose concentrations in plasma, displayed mild effects on plasma HbA(1c) and peptide C concentrations, while having no effects on non-fasting glucose. On the other hand, MNA treatment considerably lowered lipid peroxidation, protein carbonylation, completely prevented impairment of endothelium-dependent vasodilatation in the aorta that was mediated entirely by NO, but failed to affect endothelial function in resistant vessels, which was mediated only partially by NO. Most importantly, chronic treatment with MNA prolonged the long-term survival of diabetic rats. In conclusion, MNA displayed a significant anti-diabetic effect that may be linked to its vasoprotective activity. PMID- 19307697 TI - Preserved coronary arteriolar dilatation in patients with type 2 diabetes mellitus: implications for reactive oxygen species. AB - Type 2 diabetes mellitus is associated with clustering of cardiovascular risk factors that may greatly increase individuals' risk of developing coronary artery disease. Type 2 diabetes is believed to impair coronary function. However, its impact on the vasomotor function of coronary resistance vessels in humans is still debated. Reduced, preserved or even augmented dilations of coronary arterioles have been reported in subjects with type 2 diabetes. Interestingly, recent studies have suggested that reactive oxygen species (ROS), particularly hydrogen peroxide, may compensate for the loss of the vasodilatory function of coronary microvessels during disease development. Recent interventional clinical trials have yielded largely negative results, and there has even been some suggestion of harm caused by attempts to reduce ROS. Thus, it is possible that interference with ROS-related signaling might paradoxically temper the function of coronary microvessels, predisposing patients to myocardial ischemia. In this review, we aim to highlight current findings supporting a potential role for ROS in preserving coronary arteriolar dilation in type 2 diabetes mellitus. PMID- 19307698 TI - Novel approaches to improving endothelium-dependent nitric oxide-mediated vasodilatation. AB - Endothelial dysfunction, which is defined by decreased endothelium-dependent vasodilatation, is associated with an increased number of cardiovascular events. Nitric oxide (NO) bioavailability is reduced by altered endothelial signal transduction or increased formation of radical oxygen species reacting with NO. Endothelial dysfunction is therapeutically reversible and physical exercise, calcium channel blockers, angiotensin converting enzyme inhibitors, and angiotensin receptor antagonists improve flow-evoked endothelium-dependent vasodilation in patients with hypertension and diabetes. We have investigated three different approaches, with the aim of correcting endothelial dysfunction in cardiovascular disease. Thus, (1) we evaluated the effect of a cell permeable superoxide dismutase mimetic, tempol, on endothelial dysfunction in small arteries exposed to high pressure, (2) investigated the endothelial signal transduction pathways involved in vasorelaxation and NO release induced by an olive oil component, oleanolic acid, and (3) investigated the role of calcium activated K channels in the release of NO induced by receptor activation. Tempol increases endothelium-dependent vasodilatation in arteries from hypertensive animals most likely through the lowering of radical oxygen species, but other mechanisms also appear to contribute to the effect. While oleanolic acid leads to the release of NO by calcium-independent phosphorylation of endothelial NO synthase, endothelial calcium-activated K channels and an influx of calcium play an important role in G-protein coupled receptor-evoked release of NO. Thus, all three approaches increase bioavailability of NO in the vascular wall, but it remains to be addressed whether these actions have any direct benefit at a clinical level. PMID- 19307699 TI - Effect of chronic apocynin treatment on nitric oxide and reactive oxygen species production in borderline and spontaneous hypertension. AB - The purpose of this study was to investigate the effect of NAD(P)H oxidase inhibitor - apocynin (4-hydroxy-3-methoxyacetophenone) on the increase of systolic blood pressure (SBP) in borderline (BHR) and spontaneously hypertensive rats (SHR). Young 6-week-old male BHR (offspring of SHR dams and Wistar Kyoto sires) and SHR were treated with apocynin (30 mg/kg/day) for six weeks. SBP was measured by tail-cuff plethysmography. Nitric oxide synthase (NOS) activity was determined in the left ventricle and aorta. Protein expression of nuclear factor kappa B (NF-kappaB) and NAD(P)H oxidase subunits p67phox and p22phox as well as concentration of cGMP were determined for the left ventricle. Apocynin significantly decreased SBP in all groups investigated. Administration of apocynin had no effect on NOS activity in either tissue studied. However, apocynin decreased protein expression of NF-kappaB (p65) and NAD(P)H oxidase subunit p22phox in both hypertensive groups and p67phox subunit in the SHR group. Moreover, apocynin was able to prevent a decrease in cGMP concentration in the left ventricle of both hypertensive groups. In conclusion, our study demonstrated that apocynin treatment partially prevented SBP rise in borderline and spontaneously hypertensive rats, yet without increasing activity of NOS in the left ventricle and aorta. However, apocynin was able to decrease production of reactive oxygen species in hypertensive rats; thus preventing the decrease in cGMP formation. PMID- 19307700 TI - Mitochondria and vascular pathology. AB - Functional and structural changes in mitochondria are caused by the opening of the mitochondrial permeability transition pore (PTP) and by the mitochondrial generation of reactive oxygen species (ROS). These two processes are linked in a vicious cycle that has been extensively documented in ischemia/reperfusion injuries of the heart, and the same processes likely contribute to vascular pathology. For instance, the opening of the PTP causes cell death in isolated endothelial and vascular smooth muscle cells. Indeed, atherosclerosis is exacerbated when mitochondrial antioxidant defenses are hampered, but a decrease in mitochondrial ROS formation reduces atherogenesis. Determining the exact location of ROS generation in mitochondria is a relevant and still unanswered question. The respiratory chain is generally believed to be a main site of ROS formation. However, several other mitochondrial components likely contribute to ROS generation. Recent reports highlight the relevance of monoamine oxidases (MAO) and p66(Shc). For example, the absence of p66(Shc) in hypercholesterolemic mice has been reported to reduce the occurrence of foam cells and early atherogenic lesions. On the other hand, MAO inhibition has been shown to reduce oxidative stress in many cell types eliciting significant protection from myocardial ischemia. In conclusion, evidence will be presented to demonstrate that (i) mitochondria are major sites of ROS formation; (ii) an increase in mitochondrial ROS formation and/or a decrease in mitochondrial antioxidant defenses exacerbate atherosclerosis; and (iii) mitochondrial dysfunction is likely a relevant mechanism underlying several risk factors (i.e., diabetes, hyperlipidemia, hypertension) associated with atherosclerosis. PMID- 19307701 TI - Mitochondrial biogenesis in cardiac pathophysiology. AB - Cardiac performance depends on a fine balance between the work the heart has to perform to satisfy the needs of the body and the energy that it is able to produce. Thus, energy production by oxidative metabolism, the main energy source of the cardiac muscle, has to be strictly regulated to adapt to cardiac work. Mitochondrial biogenesis is the mechanism responsible for mitochondrial component synthesis and assembly. This process controls mitochondrial content and thus correlates with energy production that, in turn, sustains cardiac contractility. Mitochondrial biogenesis should be finely controlled to match cardiac growth and cardiac work. When the heart is subjected to an increase in work in response to physiological and pathological challenges, it adapts by increasing its mass and expressing a new genetic program. In response to physiological stimuli such as endurance training, mitochondrial biogenesis seems to follow a program involving increased cardiac mass. But in the context of pathological hypertrophy, the modifications of this mechanism remain unclear. What appears clear is that mitochondrial biogenesis is altered in heart failure, and the imbalance between cardiac work demand and energy production represents a major factor in the development of heart failure. PMID- 19307702 TI - Molecular determinants of heart failure with normal left ventricular ejection fraction. AB - In population-based studies, heart failure with normal left ventricular (LV) ejection fraction (HFNEF) is now increasingly recognized and referred to as diastolic heart failure. However, the pathogenic mechanisms underlying HFNEF are incompletely understood, mainly because of limited availability of human myocardial biopsy material. Nevertheless, recent studies have examined in vivo hemodynamics, in vitro cardiomyocyte function, myofilamentary protein composition, collagen content and deposition of advanced glycation end products from LV endomyocardial biopsies. These measures were compared between HFNEF patients, subjects without symptoms of heart failure (controls), patients with heart failure and reduced ejection function (HFREF), and patients with HFNEF and HFREF with diabetes mellitus. This article summarizes the various findings of these studies and focuses on the possible correlations among altered LV myocardial structure, cardiomyocyte function, myofilamentary proteins, and extracellular matrices. These findings revealed novel mechanisms responsible for diastolic LV dysfunction, and they have important therapeutic implications, particularly HFNEF, for which a specific heart failure treatment strategy is largely lacking. PMID- 19307703 TI - Functional characterization of the cAMP-binding proteins Epac in cardiac myocytes. AB - The cyclic AMP (cAMP)-binding proteins, Epac, are guanine nucleotide exchange factors for the Ras-like small GTPases. Since their discovery in 1998 and with the development of specific Epac agonists, many data in the literature have illustrated their critical role in multiple cellular events mediated by the second messenger cAMP. Given the importance of cAMP in cardiovascular physiology and physiopathology, there is a growing interest to delineate the role of these multi-domain Epac in the cardiovascular system. This review will focus on recent pharmacological and biochemical studies aiming at understanding the role of Epac in cardiomyocyte signaling and hypertrophy. PMID- 19307704 TI - Anthracycline-induced cardiotoxicity: overview of studies examining the roles of oxidative stress and free cellular iron. AB - The risk of cardiotoxicity is the most serious drawback to the clinical usefulness of anthracycline antineoplastic antibiotics, which include doxorubicin (adriamycin), daunorubicin or epirubicin. Nevertheless, these compounds remain among the most widely used anticancer drugs. The molecular pathogenesis of anthracycline cardiotoxicity remains highly controversial, although the oxidative stress-based hypothesis involving intramyocardial production of reactive oxygen species (ROS) has gained the widest acceptance. Anthracyclines may promote the formation of ROS through redox cycling of their aglycones as well as their anthracycline-iron complexes. This proposed mechanism has become particularly popular in light of the high cardioprotective efficacy of dexrazoxane (ICRF-187). The mechanism of action of this drug has been attributed to its hydrolytic transformation into the iron-chelating metabolite ADR-925, which may act by displacing iron from anthracycline-iron complexes or by chelating free or loosely bound cellular iron, thus preventing site-specific iron-catalyzed ROS damage. However, during the last decade, calls for the critical reassessment of this "ROS and iron" hypothesis have emerged. Numerous antioxidants, although efficient in cellular or acute animal experiments, have failed to alleviate anthracycline cardiotoxicity in clinically relevant chronic animal models or clinical trials. In addition, studies with chelators that are stronger and more selective for iron than ADR-925 have also yielded negative or, at best, mixed outcomes. Hence, several lines of evidence suggest that mechanisms other than the traditionally emphasized "ROS and iron" hypothesis are involved in anthracycline-induced cardiotoxicity and that these alternative mechanisms may be better bases for designing approaches to achieve efficient and safe cardioprotection. PMID- 19307705 TI - Urocortin: a protective peptide that targets both the myocardium and vasculature. AB - The urocortins are a family of endogenously produced peptide hormones that show great promise as potential drugs for the treatment of heart disease. They can increase contractility and cardiac output without causing changes in mean arterial blood pressure. As expected, the receptor for these peptides is present in cardiomyocytes, and they can bind and protect these cells from simulated ischemia and reperfusion in vitro. The receptor is present, however, in much higher density in the endothelial cells that form a continuous lining of the coronary vasculature. Functionally, the urocortin peptides have been shown to have potent local vasodilatory effects, and may affect other aspects of vascular function. In this review, we will attempt to distinguish the "cardio" from the "vascular" effects of urocortin and its homologues, including the archetypal family member, corticotrophin releasing hormone. PMID- 19307706 TI - Rabbit erythrocytes release ATP and dilate skeletal muscle arterioles in the presence of reduced oxygen tension. AB - In skeletal muscle, oxygen (O(2)) delivery to appropriately meet metabolic need requires mechanisms for detection of the magnitude of O(2) demand and the regulation of O(2) delivery. Erythrocytes, when exposed to a decrease in O(2) tension, release both O(2) and the vasodilator adenosine triphosphate (ATP). The aims of this study were to establish that erythrocytes release ATP in response to reduced O(2) tension and determine if erythrocytes are necessary for the dilation of isolated skeletal muscle arterioles exposed to reduced extraluminal O(2) tension. Rabbit erythrocytes exposed to reduced O(2) tension in a tonometer (n = 5, pO(2) = 27 +/- 3, p < 0.01) released ATP in response to reduced O(2) tension. ATP release increased in proportion to the decrease in O(2) tension. The contribution of erythrocytes to the response of skeletal muscle arterioles to reduced extraluminal O(2) tension was determined using isolated hamster cheek pouch retractor muscle arterioles perfused with buffer (n = 11, mean diameter 52 +/- 3 mum) in the absence and presence of rabbit erythrocytes. Without erythrocytes, arterioles did not dilate when exposed to reduced extraluminal O(2) tension (pO(2) = 32 +/- 4 mmHg). In contrast, when rabbit erythrocytes were present in the perfusate (hematocrit 15%), the same decrease in O(2) tension resulted in a 20 +/- 4% dilation (p < 0.01). These results provide support for the hypothesis that erythrocytes, via their ability to release O(2) along with ATP in response to exposure to reduced O(2) tension, can participate in the matching of O(2) delivery with metabolic need in skeletal muscle. PMID- 19307711 TI - Structure, modifications and ligand-binding properties of rat profilin 2a. AB - Profilins are key regulators of the actin microfilament system and in neuronal tissues the profilin 2a isoform is the most abundant and important profilin. The high-resolution crystal structure of rat profilin 2a has been determined in the absence of ligands. By comparing the structure with those of peptide-liganded profilin 2a and unliganded profilin 2b, it can be concluded that the binding site for proline-rich peptides is pre-organized. The C-terminus of profilin 2a is also well ordered in the absence of ligand peptide, in contrast to the 2b isoform which is generated by alternative splicing. Covalent modifications of four cysteine residues were also detected in profilin 2a, as well as a number of other modifications in profilin 2 from rat brain; such modifications could significantly affect the function of profilin. It was also shown that profilin 2a binds to the neuronal protein palladin, including a synthetic palladin peptide; peptides from another profilin ligand, dynamin 1, failed to interact with both profilin 1 and profilin 2a. These results allow a better understanding of the structure-function relationships and ligand binding of mammalian profilin 2a. PMID- 19307712 TI - Mycobacterium tuberculosis pantothenate kinase: possible changes in location of ligands during enzyme action. AB - The crystal structures of complexes of Mycobacterium tuberculosis pantothenate kinase with the following ligands have been determined: (i) citrate; (ii) the nonhydrolysable ATP analogue AMPPCP and pantothenate (the initiation complex); (iii) ADP and phosphopantothenate resulting from phosphorylation of pantothenate by ATP in the crystal (the end complex); (iv) ATP and ADP, each with half occupancy, resulting from a quick soak of crystals in ATP (the intermediate complex); (v) CoA; (vi) ADP prepared by soaking and cocrystallization, which turned out to have identical structures, and (vii) ADP and pantothenate. Solution studies on CoA binding and catalytic activity have also been carried out. Unlike in the case of the homologous Escherichia coli enzyme, AMPPCP and ADP occupy different, though overlapping, locations in the respective complexes; the same is true of pantothenate in the initiation complex and phosphopantothenate in the end complex. The binding site of MtPanK is substantially preformed, while that of EcPanK exhibits considerable plasticity. The difference in the behaviour of the E. coli and M. tuberculosis enzymes could be explained in terms of changes in local structure resulting from substitutions. It is unusual for two homologous enzymes to exhibit such striking differences in action. Therefore, the results have to be treated with caution. However, the changes in the locations of ligands exhibited by M. tuberculosis pantothenate kinase are remarkable and novel. PMID- 19307713 TI - Use of an in-house approach to study the three-dimensional structures of various outer membrane proteins: structure of the alcaligin outer membrane transporter FauA from Bordetella pertussis. AB - Bordetella pertussis is the bacterial agent of whooping cough in humans. Under iron-limiting conditions, it produces the siderophore alcaligin. Released to the extracellular environment, alcaligin chelates iron, which is then taken up as a ferric alcaligin complex via the FauA outer membrane transporter. FauA belongs to a family of TonB-dependent outer membrane transporters that function using energy derived from the proton motive force. Using an in-house protocol for membrane protein expression, purification and crystallization, FauA was crystallized in its apo form together with three other TonB-dependent transporters from different organisms. Here, the protocol used to study FauA is described and its three dimensional structure determined at 2.3 A resolution is discussed. PMID- 19307714 TI - Structure analysis of the conserved methyltransferase domain of human trimethylguanosine synthase TGS1. AB - Methyltransferases play an important role in the post-transcriptional maturation of most ribonucleic acids. The modification of spliceosomal UsnRNAs includes N2 dimethylation of the m(7)G cap catalyzed by trimethylguanosine synthase 1 (TGS1). This 5'-cap hypermethylation occurs during the biogenesis of UsnRNPs as it initiates the m(3)G cap-dependent nuclear import of UsnRNPs. The conserved methyltransferase domain of human TGS1 has been purified, crystallized and the crystal structure of this domain with bound substrate m(7)GpppA was solved by means of multiple-wavelength anomalous dispersion. Crystal structure analysis revealed that m(7)GpppA binds via its adenosine moiety to the structurally conserved adenosylmethionine-binding pocket, while the m(7) guanosine remains unbound. This unexpected binding only occurs in the absence of AdoMet and suggests an incomplete binding pocket for the m(7)G cap which is caused by the N terminal truncation of the protein. These structural data are consistent with the finding that the crystallized fragment of human TGS1 is catalytically inactive, while a fragment that is 17 amino acids longer exhibits activity. PMID- 19307715 TI - Structural improvement of unliganded simian immunodeficiency virus gp120 core by normal-mode-based X-ray crystallographic refinement. AB - The envelope protein gp120/gp41 of simian and human immunodeficiency viruses plays a critical role in viral entry into host cells. However, the extraordinarily high structural flexibility and heavy glycosylation of the protein have presented enormous difficulties in the pursuit of high-resolution structural investigation of some of its conformational states. An unliganded and fully glycosylated gp120 core structure was recently determined to 4.0 A resolution. The rather low data-to-parameter ratio limited refinement efforts in the original structure determination. In this work, refinement of this gp120 core structure was carried out using a normal-mode-based refinement method that has been shown in previous studies to be effective in improving models of a supramolecular complex at 3.42 A resolution and of a membrane protein at 3.2 A resolution. By using only the first four nonzero lowest-frequency normal modes to construct the anisotropic thermal parameters, combined with manual adjustments and standard positional refinement using REFMAC5, the structural model of the gp120 core was significantly improved in many aspects, including substantial decreases in R factors, better fitting of several flexible regions in electron density maps, the addition of five new sugar rings at four glycan chains and an excellent correlation of the B-factor distribution with known structural flexibility. These results further underscore the effectiveness of this normal mode-based method in improving models of protein and nonprotein components in low resolution X-ray structures. PMID- 19307716 TI - Application of molecular replacement to protein powder data from image plates. AB - Macromolecular structures can be solved via molecular replacement from powder diffraction data collected not only on multi-analyzer diffractometers but also on image plates. Diffraction peaks recorded on image plates are generally broader than those collected using an array of crystal analyzer detectors, but the image plate data often allow the use of powder data to lower d-spacings. Owing to the high incidence of overlaps in powder patterns, which is especially evident for larger structures, a multi-pattern Pawley refinement is necessary in order to distinguish intensity peaks. This work utilized various salt concentrations to produce small lattice distortions, which resulted in shifts of Bragg peak positions, in a suite of five powder patterns. Using reflection structure factors obtained from this combined refinement, the structure of hen egg-white lysozyme was determined by molecular replacement using the 60% identical human lysozyme (PDB code 1lz1) as the search model. This work also expands upon previous work by presenting a full-scale multi-species analysis combined with an investigation of the sensitivity with regard to discrimination between incorrect fold types. To test the limits of this technique, extension to higher molecular-weight structures is ongoing. PMID- 19307717 TI - Structure of Thermotoga maritima TM0439: implications for the mechanism of bacterial GntR transcription regulators with Zn2+-binding FCD domains. AB - The GntR superfamily of dimeric transcription factors, with more than 6200 members encoded in bacterial genomes, are characterized by N-terminal winged helix DNA-binding domains and diverse C-terminal regulatory domains which provide a basis for the classification of the constituent families. The largest of these families, FadR, contains nearly 3000 proteins with all-alpha-helical regulatory domains classified into two related Pfam families: FadR_C and FCD. Only two crystal structures of FadR-family members, those of Escherichia coli FadR protein and LldR from Corynebacterium glutamicum, have been described to date in the literature. Here, the crystal structure of TM0439, a GntR regulator with an FCD domain found in the Thermotoga maritima genome, is described. The FCD domain is similar to that of the LldR regulator and contains a buried metal-binding site. Using atomic absorption spectroscopy and Trp fluorescence, it is shown that the recombinant protein contains bound Ni(2+) ions but that it is able to bind Zn(2+) with K(d) < 70 nM. It is concluded that Zn(2+) is the likely physiological metal and that it may perform either structural or regulatory roles or both. Finally, the TM0439 structure is compared with two other FadR-family structures recently deposited by structural genomics consortia. The results call for a revision in the classification of the FadR family of transcription factors. PMID- 19307718 TI - Reducing radiation damage in macromolecular crystals at synchrotron sources. AB - A new strategy is presented to reduce primary X-ray damage in macromolecular crystallography. The strategy is based on separating the diffracting and damaged regions as much as feasible. The source of the radiation damage to macromolecular crystals is from two primary mechanisms: the direct excitations of electrons by absorption, and inelastic scattering of the X-rays. The first produces photoelectrons with their accompanying Auger electrons from relaxation of the core hole and the second creates Compton electrons. The properties of these two mechanisms and calculations of primary X-ray damage quantify how to modify the spatial distribution of X-rays to reduce the deleterious effects of radiation damage. By focusing the incident X-rays into vertical stripes, it is estimated that the survival (the time during which quality diffraction data can be obtained with a given X-ray flux) of large crystals can be increased by at least a factor of 1.6, while for very small platelet crystals the survival can be increased by up to a factor of 14. PMID- 19307719 TI - Tryptophan as a three-way switch in regulating the function of the secretory signalling glycoprotein (SPS-40) from mammary glands: structure of SPS-40 complexed with 2-methylpentane-2,4-diol at 1.6 A resolution. AB - The 40 kDa secretory signalling glycoprotein (SPS-40) is the first example with Trp78 in three functional orientations: (i) a resting state with a pinched conformation, (ii) a stacked conformation when bound to hexasaccharide and (iii) an obstructive conformation when inhibited by 2-methylpentane-2,4-diol (MPD). Trp78 is present in the core of the sugar-binding groove. The hexasaccharide N acetylglucosamine (GlcNAc(6)) has been shown to bind to SPS-40. As a result of this, the conformation of Trp78 alters from the native pinched conformation (chi(1) = -65.5 degrees , chi(2,1) = -78.8 degrees , chi(2,2) = 97.5 degrees ) to the stacked conformation (chi(1) = -170.0 degrees , chi(2,1) = -114.3 degrees , chi(2,2) = 61.6 degrees ). Further binding experiments showed that saccharide binding does not occur in the presence of 20% MPD. The crystal structure determination of the complex of SPS-40 with MPD revealed the presence of two MPD molecules in the sugar-binding groove. The very tightly bound MPD molecules at subsites -2 and -1 induced an unexpected and a rarely observed conformation of Trp78 (chi(1) = 55.9 degrees , chi(2,1) = 90.2 degrees , chi(2,2) = -88.9 degrees ) which is termed an obstructive conformation. The binding of MPD molecules also twisted the side chains of Glu269 and Ile272 considerably. These residues are also part of the sugar-binding groove. The observed obstructive conformation of the side chain of Trp78 in the present structure is the exact opposite of the stacked conformation. This rarely observed conformation is stabilized by a number of hydrogen bonds between Trp78 and Asn79 through water molecules W49, W229, W269, W547 and W557. PMID- 19307720 TI - On the possibility of using polycrystalline material in the development of structure-based generic assays. AB - The discovery of ligands that bind specifically to a targeted protein benefits from the development of generic assays for high-throughput screening of a library of chemicals. Protein powder diffraction (PPD) has been proposed as a potential method for use as a structure-based assay for high-throughput screening applications. Building on this effort, powder samples of bound/unbound states of soluble hen-egg white lysozyme precipitated with sodium chloride were compared. The correlation coefficients calculated between the raw diffraction profiles were consistent with the known binding properties of the ligands and suggested that the PPD approach can be used even prior to a full description using stereochemically restrained Rietveld refinement. PMID- 19307721 TI - Structure of the diaminopimelate epimerase DapF from Mycobacterium tuberculosis. AB - The meso (or D,L) isomer of diaminopimelic acid (DAP), a precursor of L-lysine, is a key component of the pentapeptide linker in bacterial peptidoglycan. While the peptidoglycan incorporated in the highly complex cell wall of the pathogen Mycobacterium tuberculosis structurally resembles that of Escherichia coli, it is unique in that it can contain penicillin-resistant meso-DAP-->meso-DAP linkages. The interconversion of L,L-DAP and meso-DAP is catalysed by the DAP epimerase DapF, a gene product that is essential in M. tuberculosis. Here, the crystal structure of the ligand-free form of M. tuberculosis DapF (MtDapF) refined to a resolution of 2.6 A is reported. MtDapF shows small if distinct deviations in secondary structure from the two-domain alpha/beta-fold of the known structures of Haemophilus influenzae DapF and Bacillus anthracis DapF, which are in line with its low sequence identity (4%, a symptom index (SI) >or=50% or a positive PPI test in patients without erosive esophagitis. Patients without such findings were classified as FH. RESULTS: Thirty-six patients had erosive reflux disease (ERD), 36 had NERD, and 23 had FH. The proportion of males was significantly higher in ERD than in FH. Atypical symptoms and IBS were more prevalent in FH than in ERD. Anxiety was more prevalent in FH than in NERD. The prevalence of pathologic acid reflux, a positive SI and a positive PPI test was similar between ERD and NERD patients. In the ERD and NERD groups, depression was independently associated with nonresponsiveness to PPIs. CONCLUSIONS: FH is a different entity from ERD or NERD, particularly in terms of gender, acid reflux patterns, psychological profiles, and the responsiveness to PPIs. PMID- 19307736 TI - Selective internal radiotherapy with Yttrium-90 microspheres for hepatic metastatic neuroendocrine tumors: a prospective single center study. AB - BACKGROUND: To assess prospectively the safety and efficacy of Yttrium-90 microspheres in patients with unresectable liver metastases from neuroendocrine tumors. MATERIALS AND METHODS: Microspheres were administered via a temporarily placed hepatic catheter. Patients were monitored prospectively. All patients were followed with laboratory and imaging studies at regular intervals to determine response rates. Toxicity and quality of life scores were measured. RESULTS: Nine patients (7 female) with a mean age of 58.8 years were enrolled in this prospective trial. The mean tumor load was 58.8%. The estimated percentage shunting to the lungs on MAA scans was 5.04 +/- 2.4%. Visceral artery embolization of extrahepatic arteries before treatment was performed in 6 patients. The median dose of microspheres was 2.1 +/- 0.4 GBq. A total of 12 therapy sessions was performed. The mean follow-up was 21.7 months. Technical success was 100%. No major complications occurred. Survival rates were 100, 57 and 57% for 1, 2 and 3 years, respectively. Three months after SIRT therapy partial response (PR) was seen in 6 patients (66%). Calculated reduction of liver metastasis volume was 49%. In 3 patients (33%) stable disease was seen with a calculated tumor reduction of 13%. The estimated time to progression was 11.1 months. CONCLUSION: Radioembolization with (90)Y microspheres is safe and produces high response rates even with extensive tumor replacement for up to 1 year. Acute and late toxicity was very low. Further investigations compared with other local ablative techniques is warranted. PMID- 19307737 TI - Cardiac toxicity with capecitabine, vinorelbine and trastuzumab therapy: case report and review of fluoropyrimidine-related cardiotoxicity. AB - A 45-year-old woman presented with a metastatic breast carcinoma and was treated with capecitabine, oral vinorelbine and trastuzumab combination therapy. The initial echocardiogram and the ECG were considered normal. She began treatment with 3-weekly cycles of the combination therapy. After the fourth dose of capecitabine, she presented with severe chest and arm pain, which was responsive to nitroglycerine spray. ECG at admission demonstrated tachycardia with ST segment elevation suggesting ischemia. The clinical symptoms returned to baseline after a few hours and within 24 h the ECG showed inverted T in leads V3-V6. Cardiac ultrasonography revealed hypokinesia in the left ventricle without segmentary hypokinesia, with mildly reduced global systolic function, which normalized 1 week later. Two weeks later, she was rechallenged with capecitabine. After the fourth dose, the patient developed chest pain. ECG showed infero-apico lateral injury, which normalized after administration of nitrates, nicorandil and verapamil and discontinuation of capecitabine. This case suggests that capecitabine can lead to the cardiotoxicity characteristic of other fluoropyrimidines. Therefore, it is important to inform patients about the risk of angina-like chest pain, to stop treatment immediately if symptoms occur, and to monitor the patient in hospital. Fluoropyrimidine rechallenge should be avoided because of the risk of ischemic event or sudden death. PMID- 19307738 TI - A prospective, multicenter, phase 2 study of imatinib mesylate in korean patients with metastatic or unresectable gastrointestinal stromal tumor. AB - OBJECTIVES: This prospective, multicenter, phase 2 study evaluated the efficacy and safety of imatinib mesylate and assessed KIT and PDGFRA gene mutation status in Korean patients with gastrointestinal stromal tumors (GISTs). METHODS: Forty seven patients with pathologically proven KIT-positive metastatic or unresectable GISTs were accrued from eight institutions in Korea. Imatinib was administered orally at 400 mg once daily. In case of disease progression, the dose was escalated to 600 mg once daily, then 400 mg twice daily. KIT and PDGFRA mutations were analyzed in 29 of the 47 patients. RESULTS: Imatinib produced partial responses in 30 patients (63.8%; 95% confidence interval, 50.1-77.6%) and stable disease in 13 patients (27.7%). The median time to response was 2.6 months (range, 1.0-6.2 months). With a median follow-up of 62 months (range, 32-67 months), 4-year progression-free survival and overall survival rates were 50 and 65%, respectively. The most common adverse events were anemia, neutropenia, edema, and skin rash (predominantly of grade 1-2). There were no treatment related deaths. In the subset evaluated for mutational status, 24 patients (82.8%) had KIT exon 11 mutations and 1 (3.4%) had a KIT exon 9 mutation. CONCLUSIONS: Imatinib is effective and safe in Korean patients with metastatic or unresectable GIST. PMID- 19307739 TI - Fixed-dose-rate gemcitabine in combination with oxaliplatin in patients with metastatic pancreatic cancer refractory to standard-dose-rate gemcitabine: a single-institute study. AB - OBJECTIVES: There is a paucity of data exploring treatment options for refractory pancreatic cancer. Oxaliplatin has interesting activity in second-line therapy. Fixed-dose-rate gemcitabine (GFDR, 10 mg/m(2)/min) has shown promising results in patients with advanced pancreatic cancer over standard-dose-rate (30 min) gemcitabine (GSDR). METHODS: We conducted a retrospective analysis of the experience of our cancer center with GFDR and oxaliplatin (GEMOX) in patients who failed GSDR. GEMOX consisted of gemcitabine 1,000 mg/m(2) over 100 min on day 1 and oxaliplatin 100 mg/m(2) over 2 h on day 2 every 2 weeks. Eligible patients were required to have measurable metastatic adenocarcinoma of the pancreas and to have failed prior GSDR. RESULTS: Seventeen patients (median age 62 years) who were treated at the Ohio State University with GEMOX following GSDR failure between November 2003 and January 2008 were included in this study. Twenty-four percent of all patients had a partial response, 29% had stable disease and 47% had progressive disease. The median progression-free survival was 2.6 months and the median overall survival was 6.4 months. There were no unexpected toxicities. CONCLUSION: GEMOX shows interesting activity and acceptable tolerability in patients with metastatic pancreatic cancer who failed prior GSDR. Our results are consistent with previously published results. PMID- 19307740 TI - Phase I/II study of twenty-four-hour infusion of irinotecan in combination with oral UFT plus leucovorin for metastatic colorectal cancer. AB - OBJECTIVE: Irinotecan has, in general, been administered as a 90-min infusion. However, several studies have demonstrated that continuous infusion seems to be a promising method of delivering irinotecan. This phase I/II trial was performed to evaluate the efficacy and safety of continuous infusion of irinotecan combined with UFT plus leucovorin (LV) for metastatic colorectal cancer. METHODS: Escalating doses of irinotecan (90-110 mg/m(2)) were administered by 24-hour infusion on day 1. UFT 300 mg/m(2)/day and LV 75 mg/day were administered orally, in 3 divided daily doses, on days 3-7 and 10-14. The treatment cycles were repeated every 2 weeks. RESULTS: In the phase I study, the maximum tolerated dose of irinotecan was 110 mg/m(2) and the recommended dose for the phase II study was determined to be 100 mg/m(2). Thirty-six patients, including 3 patients at the recommended dose in the phase I study, were evaluated in the phase II study. The common grade 3/4 toxicities were leucopenia, neutropenia, diarrhea and anorexia. The response rate was 63.9%, and the median progression-free and overall survival times were 8.3 and 24.6 months, respectively. CONCLUSION: A 24-hour infusion of irinotecan combined with UFT/LV is feasible and active for metastatic colorectal cancer. PMID- 19307741 TI - High orotate phosphoribosyltransferase gene expression predicts complete response to chemoradiotherapy in patients with squamous cell carcinoma of the esophagus. AB - OBJECTIVE: Chemoradiotherapy (CRT) is a possible alternative to surgery for esophageal cancer. As complete response (CR) to CRT is essential for a good prognosis, potential biomarkers predictive of CR were explored. METHODS: Endoscopic tumor biopsies were obtained from 41 patients with stage II-III esophageal squamous cell carcinoma before 5-fluorouracil/cisplatin-based definitive CRT. cDNA was derived from RNA isolated from microdissected tumor cells. mRNA expression levels of 10 genes involved in CRT or tumor biology were measured using quantitative real-time RT-PCR. RESULTS: Expression levels of orotate phosphoribosyltransferase (OPRT) and dihydrofolate reductase mRNA were significantly higher in the CR group compared with the non-CR group (p = 0.0206 and 0.0191, respectively). Matrix metalloproteinase 9 mRNA expression was significantly lower in the CR group (p = 0.0436). CR rates were significantly higher in patients with node-negative disease and high expression levels of OPRT and dihydrofolate reductase genes (p = 0.0448, 0.0104 and 0.0104, respectively). No significant difference in CR rates was observed for other variables. Multivariate analysis revealed that high OPRT gene expression was an independent predictive factor of CR (p = 0.0192). It was also significantly associated with good prognosis (p = 0.0450). CONCLUSION: High OPRT gene expression may be a predictive factor of CR to 5-fluorouracil/cisplatin-based CRT in esophageal cancer. PMID- 19307742 TI - Modulation of glycine receptor function by the synthetic cannabinoid HU210. AB - Loss of inhibitory synaptic transmission within the dorsal horn of the spinal cord plays a key role in the development of chronic pain following inflammation or nerve injury. Inhibitory postsynaptic transmission in the adult spinal cord involves mainly glycine. HU210 is a non-psychotropic, synthetic cannabinoid. As we hypothesized that non-CB receptor mechanisms of HU210 might contribute to its anti-inflammatory and anti-nociceptive effects we investigated the interaction of HU210 with strychnine-sensitive alpha(1 )glycine receptors by using the whole cell patch clamp technique. HU210 showed a positive allosteric modulating effect in a low micromolar concentration range (EC(50): 5.1 +/- 2.6 micromol/l). Direct activation of glycine receptors was observed at higher concentrations above 100 micromol/l (EC(50): 188.7 +/- 46.2 micromol/l). These in vitro results suggest that strychnine-sensitive glycine receptors may be a target for HU210 mediating some of its anti-inflammatory and anti-nociceptive properties. PMID- 19307743 TI - Cavernous malformations of the central nervous system in the pediatric age group. AB - OBJECTIVE: The main clinico-diagnostic features, risk factors and associated diseases of cavernous malformations (CMs), also called cavernous angiomas or cavernomas, of the central nervous system (CNS) in children are described, and the most relevant differences compared to the affected adult population are pointed out, focusing on the management of pediatric patients harboring cranial and spinal CMs. MATERIALS: This was a retrospective study of a series of 42 children symptomatic for CMs of the cranial and spinal compartments (35 supratentorial brain lesions, 5 infratentorial and 2 in the spinal region) operated on between 1975 and 2005, with a clinical follow-up ranging from 12 to 192 months. The results were compared with those found in the most recent literature dealing with pediatric CMs of the CNS. RESULTS: Surgical treatment produced excellent or good results in 69% of our 42 children. Unchanged neurological deficits were observed in 23.8% of cases, while morbidity from surgical procedures was 7.14%. Mortality was absent in this series. These surgical results are comparable with those found in the literature, where morbidity and mortality rates from surgery are 8.8 and 1.13%, respectively, and are mostly associated with procedures for the excision of deep, critically located cavernomas. CONCLUSION: CMs represent the most common CNS vascular lesion in children, although their incidence is 4 times lower than that of the adult population. The natural history of pediatric CMs throughout the neuraxis seems to be more aggressive than in adult patients; these lesions have higher rates of growth and hemorrhage, larger dimensions and often atypical radiological pictures at diagnosis. Beside the familial form of the disease, which is more often associated with multiple lesions and an earlier age of clinical presentation, the major risk factor for CMs in children seems to be radiotherapy for CNS tumors. Furthermore, a greater number of CMs coexistent with mixed angiomatous lesions have been reported in children than in adults. Surgical results are related to the preoperative neurological status of the children; symptomatic patients who are operated on early, before they develop severe neurological deficits or long standing seizures, may achieve the best clinical outcome. Radiosurgery does not seem to be advisable in children as an alternative treatment for deep CMs or those causing epilepsy. PMID- 19307744 TI - Pediatric cavernous malformation in the central nervous system: report of 66 cases. AB - OBJECTIVE: To investigate the clinical features and treatment outcome of pediatric cavernous malformation (CM) in the central nervous system. PATIENTS AND METHODS: Retrospective review of clinical data and treatment outcome for 66 pediatric patients under the age of 18 treated for CM at Huashan Hospital within the past 10 years from January 1998 to December 2007. RESULTS: In our consecutive series, the mean age at onset of the CM was 11.6 years (age from 15 months to 17.8 years), and the boy-to-girl ratio was 1.54:1 (40:26). Main initial clinical manifestations in cerebral CM (CCM) included: seizures (47.7%, 31/65), headache (46.2%, 30/65), intracranial hemorrhage (20%, 13/65), and focal neurological deficits (12.3%, 8/65). There were 59 children with single CM and 7 children with multiple CMs: 55 (83.3%) with supratentorial CM, 6 (9.1%) with infratentorial CM, 4 (6.1%) with supra- and infratentorial CM, and 1 (1.5%) with intramedullary CM. Sixty-two (93.9%; 62/64) children underwent microsurgical operations, and with the help of neuronavigation (19 cases), intraoperative ultrasonography (6 cases), and neuronavigation combined with intraoperative ultrasonography (3 cases). And 4 (6.1%; 4/66) children did not undergo any surgical intervention. There was no major morbidity or mortality from surgical procedures. Follow-up data of 50 children was available with a mean follow-up time of 39.1 months (range from 5 to 112 months), including 46 operated children and 4 children managed without operation. In the 46 operated children, the overall long-term post-treatment results were satisfactory: 73.9% no sign or symptom associated with CMs, 19.6% improved obviously, only 1 (2.2%) boy with unrestored paraplegia incurred by spinal cord CM, and 2 boys (4.3%) with controllable seizures occurring after initial 5 symptom-free years (one without need of antiepileptic drugs, AEDs). Of 24 children followed-up with preoperative seizures, only 1 boy needed therapeutic AEDs and the others did not need any after resections of the CMs. Three children managed without operations (2 with headache and 1 with seizures) had spontaneous disappearance of symptoms and stable intracerebral lesions at follow-up MRI images. CONCLUSIONS: For symptomatic solitary CM, the treatment of choice is complete microsurgical excision preceded by careful anatomical and functional evaluation, and the risk of operation can be decreased to lower level with the help of neuronavigation and intraoperative ultrasonography. For multiple CM, the treatment modalities must be considered cautiously. A much longer follow-up remains mandatory for appropriate treatment strategies. PMID- 19307745 TI - Surgical treatment of myelomeningocele carried out at 'time zero' immediately after birth. AB - BACKGROUND/AIMS: To present a protocol of immediate surgical repair of myelomeningocele (MMC) after birth ('time zero') and compare this surgical outcome with the surgery performed after the newborn's admission to the nursery before the operation. METHODS: Data from the medical files of 31 patients with MMC that underwent surgery after birth and after admission at the nursery (group I) were compared with a group of 23 patients with MMC admitted and prospectively followed, who underwent surgery immediately after birth--'at time zero' (group II). RESULTS: The preoperative rupture of the MMC occurred more frequently in group I (67 vs. 39%, p < 0.05). The need for ventriculoperitoneal shunt was 84% in group I and 65% in group II and 4 of them were performed during the same anesthetic time as the immediate MMC repair, with no statistically significant difference. Group I had a higher incidence of small dehiscences when compared to group II (29 vs. 13%, p < 0.05); however, there was no statistically significant difference regarding infections. After 1 year of follow-up, 61% of group I showed neurodevelopmental delay, whereas only 35% of group II showed it. CONCLUSIONS: The surgical intervention carried out immediately after the birth showed benefits regarding a lower incidence of preoperative rupture of the MMC, postoperative dehiscences and lower incidence of neurodevelopmental delay 1 year after birth. PMID- 19307746 TI - Treatment of posthemorrhagic hydrocephalus in premature infants with subcutaneous reservoir drainage. AB - AIMS: To investigate the effectiveness of subcutaneous reservoir drainage as a treatment for the different types of posthemorrhagic hydrocephalus in premature infants. METHODS: 11 premature infants with posthemorrhagic hydrocephalus underwent subcutaneous reservoir embedment surgery, and cerebrospinal fluid (CSF) was drained via the reservoir intermittently for 8 weeks. During the period of drainage, ultrasound and computerized tomography were used to measure ventricle size. CSF examinations were performed routinely to assess the presence of intraventricular hemorrhage (IVH) and/or infection. RESULTS: (1) Five infants were diagnosed as having obstructive hydrocephalus; 2 had nearly normal ventricle sizes with treatment and drainage was stopped after 8 weeks, 2 had nearly normal ventricle sizes after 4 more weeks of drainage, and 1 underwent ventriculoperitoneal shunt (V-P shunt) surgery due to failure of ventricle size reduction. (2) Six infants were diagnosed as having communicating hydrocephalus; 4 had further enlarged ventricle size after 8 weeks and underwent V-P shunt surgery, 1 had treatment aborted at week 8 of drainage, and only 1 had moderate reduction of ventricle size after 8 weeks. (3) None of the 11 infants had progressive IVH and/or intracranial infection during drainage. CONCLUSIONS: Subcutaneous reservoir drainage is a suitable and safe treatment for posthemorrhagic hydrocephalus in premature infants. It is more effective for obstructive hydrocephalus than for communicating hydrocephalus. PMID- 19307747 TI - Effect of pioglitazone on sciatic nerve ischemia/reperfusion injury in rats. AB - OBJECTIVE: Evaluation of the effect of pioglitazone on sciatic nerve ischemia/reperfusion (I/R) injury in rat. METHOD: Sixty rats were divided into 10 groups (n = 6). Treatment groups received 15 mg/kg pioglitazone intraperitoneally 1 h before induction of I/R by clamping the right common iliac and femoral arteries for 3 h. After certain time intervals of reperfusion (0 h, 3 h, 1, 4, and 7 days), the function of the hind limb was assessed using behavioral scores based on gait, grasp, paw position, and pinch sensitivity. The sciatic nerve was removed for light microscopy studies and graded for ischemic fiber degeneration (IFD) and edema. Plasma malondialdehyde (MDA) level was measured as an indicator of lipid peroxidation at the end of reperfusion intervals. RESULT: Behavioral scores were improved in the pioglitazone groups just on the 4th and 7th days of reperfusion (p < 0.05). Comparison of the pioglitazone with the control groups showed significant differences in edema at 4 and 7 days. Although IFD decreased in the pioglitazone group at 7 days of reperfusion, it was not statistically significant. In addition, the MDA level was significantly lower in pioglitazone treated groups. CONCLUSION: Our results show the protective effect of pioglitazone on sciatic nerve I/R injury. PMID- 19307748 TI - Germinal matrix hemorrhage of prematurity: treatment approaches and outcomes in a single institutional review in the Ukraine. AB - OBJECTIVE: To review the treatment outcome of germinal matrix hemorrhage (GMH) in premature infants in a single Ukrainian institution in an effort to determine optimal diagnostic and therapeutic approaches. MATERIALS AND METHODS: Eight hundred and thirty-five premature newborns (gestational age 33.0 +/- 2.50 weeks, birth weight 2,124.81 +/- 282.54 g; mean +/- SD) were examined for the development of perinatal hypoxic-ischemia injury and asphyxia condition. This research focuses on various types of massive intracranial hemorrhage (ICH) and posthemorrhagic hydrocephalus (PHH). The diagnostic methods were based on intracranial imaging studies and clinical features that are present at birth. The therapeutic and preventive strategies consist of parental counseling, supportive and rehabilitative care for affected infants. Surgical intervention was indicated for the prevention and treatment of severe PHH. Thirty-four preterm infants were treated by ventricular-peritoneal/subgaleal shunting with close monitoring of intracerebral pressure. RESULTS: Massive GMH took place at 32-35 gestational weeks as a result of increased periventricular anastomosis. GMH was unusual in full-term newborns, whereas cerebral hypoxic-ischemic injuries were more common in full-term neonates. Approximately 98% of premature infants with low birth weight survived and 2% died due to respiratory distress syndrome and other complications. ICH which occurs in neonates at 24-28 gestational weeks was mainly due to immature vascular walls and insufficiency of vascular anastomosis at the germinal matrix. CONCLUSIONS: ICH occurring in the germinal matrix of premature newborns is closely related to the development of the brain vasculature. Evacuation of the hematoma is more detrimental than beneficial, despite the rapid strides being taken to keep low-birth weight premature infants alive. Therefore, the treatment of ICH and PHH requires a fundamental understanding of pathogenetic changes, which is necessary for the neurorehabilitation and immediate elimination of cerebral compression and its complications. PMID- 19307749 TI - Repression of TNF-alpha-induced IL-8 expression by the glucocorticoid receptor beta involves inhibition of histone H4 acetylation. AB - Increased expression of a number of proinflammatory genes, including IL-8, is associated with inflammatory conditions such as asthma. Glucocorticoid receptor (GR)beta, one of the GR isoforms, has been suggested to be upregulated in asthma associated with glucocorticoid insensitivity and to work as a dominant negative inhibitor of wild type GRalpha. However, recent data suggest that GRbeta is not a dominant negative inhibitor of GRalpha in the transrepressive process and has its own functional role. We investigated the functional role of GRbeta expression in the suppressive effect of glucocorticoids on tumor necrosis factor (TNF)-alpha induced IL-8 release in an airway epithelial cell line. GRbeta expression was induced by treatment of epithelial cells with either dexamethasone or TNF-alpha. GRbeta was able to inhibit glucocorticoid-induced transcriptional activation mediated by binding to glucocorticoid response elements (GREs). The suppressive effect of dexamethasone on TNF-alpha-induced IL-8 transcription was not affected by GRbeta overexpression, rather GRbeta had its own weak suppressive activity on TNF-alpha-induced IL-8 expression. Overall histone deacetylase activity and histone acetyltransferase activity were not changed by GRbeta overexpression, but TNF-alpha-induced histone H4 acetylation at the IL-8 promoter was decreased with GRbeta overexpression. This study suggests that GRbeta overexpression does not affect glucocorticoid-induced suppression of IL-8 expression in airway epithelial cells and GRbeta induces its own histone deacetylase activity around IL-8 promoter site. PMID- 19307750 TI - Calcitonin induces connective tissue growth factor through ERK1/2 signaling in renal tubular cells. AB - Calcitonin (CT), a polypeptide hormone, plays important roles in a variety of physiological processes. CT has been used clinically to treat osteoporosis and humoral hypercalcemia of malignancy. In order to clarify the pharmacological effects of CT in the kidney, we identified potential downstream genes induced by CT in the renal cells. Using a cDNA subtraction hybridization method, we identified connective tissue growth factor (CTGF) as a CT-induced gene in the porcine renal cell line, LLC-PK1. Furthermore, we found that CT-mediated induction of the gene was not inhibited by cycloheximide, which suggests that CTGF gene was not induced by an increased synthesis of regulating proteins. Therefore, CTGF is an immediate early gene. We further demonstrated that the regulation of CTGF gene expression by CT involved the ERK1/2 pathway, because PD98059, a MEK1 inhibitor, partially inhibited the mRNA expression of CTGF induced by CT. CT-induced CTGF protein expression was also observed in vivo. Our present findings suggest that CT induces the transcription of CTGF through ERK1/2 phosphorylation. We also identified twelve other genes induced by CT that, like CTGF, were related to wound healing. These results suggest that CT may have an effect on renal differentiation and wound healing in the kidney. PMID- 19307751 TI - Soluble mediators from mesenchymal stem cells suppress T cell proliferation by inducing IL-10. AB - Mesenchymal stem cells (MSCs) can inhibit T cell proliferation; however, the underlying mechanisms are not clear. In this study, we investigated the mechanisms of the immunoregulatory activity of MSCs on T cells. Irradiated MSCs co-cultured with either na?ve or pre-activated T cells in a mixed lymphocyte reaction (MLR) significantly suppressed T cell proliferation in a dose-dependent manner, irrespective of allogeneic disparity between responders and MSCs. Transwell assays revealed that the suppressive effect was primarily mediated by soluble factors that induced apoptosis. Splenocytes stimulated with alloantigen in the presence of the MSC culture supernatant (CS) produced a significant amount of IL-10, which was attributed to an increase in the number of IL-10 secreting cells, confirmed by an ELISPOT assay. The blockade of IL-10 and IL-10 receptor interaction by anti-IL-10 or anti-IL-10-receptor antibodies abrogated the suppressive capacity of MSC CS, indicating that IL-10 plays a major role in the suppression of T cell proliferation. The addition of 1-methyl-DL-tryptophan (1 MT), an indoleamine 2,3-dioxygenase (IDO) inhibitor, also restored the proliferative capacity of T cells. In conclusion, we demonstrated that soluble mediators from culture supernatant of MSCs could suppress the proliferation of both naive and pre-activated T cells in which IL-10 and IDO play important roles. PMID- 19307752 TI - LL-37 inhibits serum amyloid A-induced IL-8 production in human neutrophils. AB - Serum amyloid A (SAA) has been regarded as an important mediator of inflammatory responses. The effect of several formyl peptide receptor-like 1 (FPRL1) ligands on the production of IL-8 by SAA was investigated in human neutrophils. Among the ligands tested, LL-37 was found to specifically inhibit SAA-induced IL-8 production in transcriptional and post-transcriptional levels. Since SAA stimulated IL-8 production via ERK and p38 MAPK in human neutrophils, we tested the effect of LL-37 on SAA induction for these two MAPKs. LL-37 caused a dramatic inhibition of ERK and p38 MAPK activity, which is induced by SAA. LL-37 was also found to inhibit SAA-stimulated neutrophil chemotactic migration. Further, the LL 37-induced inhibitory effect was mediated by FPRL1. Our findings indicate that LL 37 is expected to be useful in the inhibition of SAA signaling and for the development of drugs against SAA-related inflammatory diseases. PMID- 19307753 TI - Alterations of epinephrine-induced gluconeogenesis in aging. AB - The effects of glucagon and epinephrine on gluconeogenesis in young (4 month) and old (24 month) Fisher 344 rat hepatocytes were compared. In contrast to glucagon, which had a similar effect on gluconeogenesis in both young and old cells, epinephrine caused a smaller increase in gluconeogenesis in old rat hepatocytes than in young hepatocytes. beta2 adrenergic receptor (beta2-AR) expression slightly decreased in aged rat liver, and there were differences between young and old hepatocytes in their patterns of G protein coupled receptor kinases, which are involved in the activation of beta2-AR receptor signal desensitization. The major isoform of the kinase changed from GRK2 to GRK3 and the expression of beta-arrestin, which is recruited by the phosphorylated beta2-AR for internalization and degradation, increased in aged rat liver. GRK3 overexpression also decreased the glucose output from young rat hepatocytes. We conclude that an age-associated reduction in epinephrine-induced gluconeogenesis occurs through the epinephrine receptor desensitizing system. PMID- 19307754 TI - ICAM-1/LFA-1 interaction contributes to the induction of endothelial cell-cell separation: implication for enhanced leukocyte diapedesis. AB - The basic route and mechanism for diapedesis has not yet to be fully defined. Here we present evidence that "cell-cell separation" between endothelial cells (ECs) may provide a route for leukocyte diapedesis. We unexpectedly found that extensive interaction between peripheral blood leukocytes and ECs that were activated by TNF-alpha induced the opening of EC contacts and, surprisingly, resulted in cell-cell separation. This event was specific to the intercellular adhesion molecules-1 (ICAM-1)/leukocyte function- associated antigen-1 interaction, as demonstrated by the following: (1) ICAM-1 expression correlated with increased EC contraction; and (2) the blocking of ICAM-1 selectively inhibited EC separation. Thus, we suggest that "cell-cell separation" could be a mechanism for diapedesis in situations that may require massive leukocyte infiltration. PMID- 19307755 TI - Chromosomal instability is more frequent in metastasized than in non-metastasized pulmonary carcinoids but is not a reliable predictor of metastatic potential. AB - Pulmonary carcinoids are infrequent neoplasms of the lung that normally display a less aggressive biological behavior compared to small cell and non-small cell lung cancers. Approximately 15-25% of carcinoids, in particular atypical carcinoids, show lymph node metastasis and have a worse prognosis than their non metastasized counterparts. To date, there is no morphological or molecular marker that may help to differentiate between carcinoids that metastasize and carcinoids of identical differentiation that show only local tumor growth. In this study, we analyzed 7 metastasized and 10 non-metastasized pulmonary carcinoids for chromosomal and microsatellite instability in order to determine whether microsatellite instability or chromosomal imbalances are associated with metastasis. Due to the rare occurrence of metastasized carcinoids we compared our results of chromosomal instability with the hitherto published comparative genomic hybridization (CGH) profiles of pulmonary carcinoids, for which information about the absence or presence of metastasis was available. While microsatellite instability was not detected we found chromosomal instability as a common event in pulmonary carcinoids with an increase of frequency and extent of chromosomal alterations in atypical and metastasized carcinoids. These findings are in accordance with the collected and herein compiled data of previous studies and indicate increasing numbers of chromosomal imbalances to play a role in the sequential process of tumor development and metastasis. PMID- 19307756 TI - Polymorphisms of COTL1 gene identified by proteomic approach and their association with autoimmune disorders. AB - To select candidate genes, we attempted to comparative analysis of protein levels between rheumatoid arthritis (RA) patients and healthy controls by two dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS). We identified 17 proteins that showed up- or down-regulated spots in RA patients. We found that coactosin-like1 (COTL1) were highly expressed in RA patients compared with healthy controls. We performed a case-control study to determine whether the COTL1 gene polymorphisms were associated with RA and systemic lupus erythematosus (SLE). The genotype frequency of c.-1124G>T and the allelic frequency of c.484G>A in RA patients, and the genotype frequency of c.484G>A in SLE patients were significantly different from healthy controls (P=0.009, 0.027, and 0.025, respectively). We also investigated the correlation with the levels of rheumatoid factor (RF) and anti cyclic citrullinated peptide (CCP) antibody in RA patients, and anti-nuclear antibodies (ANA) in SLE patients. The c.484G>A polymorphism in RA patients has significant association with the levels of anti-CCP antibody (P=0.03). Our findings demonstrated that c.-1124G>T and c. 484G>A polymorphisms of the COTL1 gene might be associated with the genetic susceptibility of autoimmune disorders. PMID- 19307757 TI - Tunicamycin enhances TRAIL-induced apoptosis by inhibition of cyclin D1 and the subsequent downregulation of survivin. AB - TNF-related apoptosis-inducing ligand (TRAIL) has been proposed as a promising cancer therapy that preferentially induces apoptosis in cancer cells, but not most normal tissues. However, many cancers are resistant to TRAIL by mechanisms that are poorly understood. In this study, we showed that tunicamycin, a naturally occurring antibiotic, was a potent enhancer of TRAIL-induced apoptosis through downregulation of survivin. The tunicamycin-mediated sensitization to TRAIL was efficiently reduced by forced expression of survivin, suggesting that the sensitization was mediated at least in part through inhibition of survivin expression. Tunicamycin also repressed expression of cyclin D1, a cell cycle regulator commonly overexpressed in thyroid carcinoma. Furthermore, silencing cyclin D1 by RNA interference reduced survivin expression and sensitized thyroid cancer cells to TRAIL; in contrast, forced expression of cyclin D1 attenuated tunicamycin-potentiated TRAIL-induced apoptosis via over-riding downregulation of survivin. Collectively, our results demonstrated that tunicamycin promoted TRAIL induced apoptosis, at least in part, by inhibiting the expression of cyclin D1 and subsequent survivin. Of note, tunicamycin did not sensitize the differentiated thyroid epithelial cells to TRAIL-induced apoptosis. Thus, combined treatment with tunicamycin and TRAIL may offer an attractive strategy for safely treating resistant thyroid cancers. PMID- 19307758 TI - Genetic and phenotypic diversity of fenitrothion-degrading bacteria isolated from soils. AB - Twenty-seven fenitrothion-degrading bacteria were isolated from different soils, and their genetic and phenotypic characteristics were investigated. Analysis of the 16S rDNA sequence showed that the isolates were related to members of the genera Burkholderia, Pseudomonas, Sphingomonas, Cupriavidus, Corynebacterium, and Arthrobacter. Among the 27 isolates, 12 different chromosomal DNA fingerprinting patterns were obtained by polymerase chain reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences. The isolates were able to utilize fenitrothion as a sole source of carbon and energy, producing 3-methyl-4- nitrophenol as the intermediate metabolite during the complete degradation of fenitrothion. Twenty-two of 27 isolates were able to degrade parathion, methyl parathion, and p-nitrophenol, but only strain BS2 could degrade EPN (O-ethyl-O-p nitrophenyl phenylphosphorothioate) as a sole source of carbon and energy for growth. Eighteen of the 27 isolates had plasmids. When analyzed with PCR amplification and dot-blotting hybridization using various specific primers targeted to the organophosphorus pesticide hydrolase genes of the previously reported isolates, none of the isolates showed positive signals, suggesting that the corresponding genes of our isolates had no significant sequence homology with those of the previously isolated organophosphate pesticide-degrading bacteria. PMID- 19307759 TI - Identification and functional characterization of an afsR homolog regulatory gene from Streptomyces venezuelae ATCC 15439. AB - Sequencing analysis of a 5-kb DNA fragment from Streptomyces venezuelae ATCC 15439 revealed the presence of one 3.1-kb open reading frame (ORF), designated afsRsv. The deduced product of afsR-sv (1,056 aa) was found to have high homology with the global regulatory protein AfsR. Homology-based analysis showed that afsR sv represents a transcriptional activator belonging to the Streptomyces antibiotic regulatory protein (SARP) family that includes an Nterminal SARP domain containing a bacterial transcriptional activation domain (BTAD), an NB-ARC domain, and a Cterminal tetratricopeptide repeat domain. Gene expression analysis by reverse transcriptase PCR (RT-PCR) demonstrated the activation of transcription of genes belonging to pikromycin production, when afsR-sv was overexpressed in S. venezuelae. Heterologous expression of the afsR-sv in different Streptomyces strains resulted in increased production of the respective antibiotics, suggesting that afsR-sv is a positive regulator of antibiotics biosynthesis. PMID- 19307760 TI - Gene cloning, purification, and characterization of a cold-adapted lipase produced by Acinetobacter baumannii BD5. AB - Acinetobacter baumannii BD5 was isolated from waters of Baek-du mountain, and the lipase gene was cloned using a PCR technique. The deduced amino acid sequence of the lipase and lipase chaperone were found to encode proteins of 325 aa and 344 aa with a molecular mass of 35 kDa and 37 kDa, respectively. The lipase gene was cloned and expressed in Escherichia coli BL21 (trxB) as an inclusion body, which was subsequently solubilized by urea, and then purified using Ni-affinity chromatography. After being purified, the lipase was refolded by incubation at 4oC in the presence of a 1:10 molar ratio of lipase:chaperone. The maximal activity of the refolded lipase was observed at a temperature of 35 degrees and pH 8.3 when p-NP caprate (C10) was used as a substrate; however, 28% of the activity observed at 35 degrees was still remaining at 0 degrees . The stability of the purified enzyme at low temperatures indicates that it is a cold-adapted enzyme. The refolded lipase was activated by Ca2+, Mg2+, and Mn2+, whereas Zn2+ and Cu2+ inhibited it. Additionally, 0.1% Tween 20 increased the lipase activity by 33%, but SDS and Triton X-100 inhibited the lipase activity by 40% and 70%, respectively. PMID- 19307761 TI - Functional expression of SAV3818, a putative TetR-family transcriptional regulatory gene from Streptomyces avermitilis, stimulates antibiotic production in Streptomyces species. AB - Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-andconstitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the lowproducer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species. PMID- 19307762 TI - Development of an analysis program of type I polyketide synthase gene clusters using homology search and profile hidden Markov model. AB - MAPSI (Management and Analysis for Polyketide Synthase Type I) has been developed to offer computational analysis methods to detect type I PKS (polyketide synthase) gene clusters in genome sequences. MAPSI provides a genome analysis component, which detects PKS gene clusters by identifying domains in proteins of a genome. MAPSI also contains databases on polyketides and genome annotation data, as well as analytic components such as new PKS assembly and domain analysis. The polyketide data and analysis component are accessible through Web interfaces and are displayed with diverse information. MAPSI, which was developed to aid researchers studying type I polyketides, provides diverse components to access and analyze polyketide information and should become a very powerful computational tool for polyketide research. The system can be extended through further studies of factors related to the biological activities of polyketides. PMID- 19307763 TI - Gene cloning, expression, and characterization of a new carboxylesterase from Serratia sp. SES-01: comparison with Escherichia coli BioHe enzyme. AB - The carboxylesterase-encoding gene (bioHs) of a newly isolated strain, Serratia sp. SES-01, was cloned from the genomic DNA library by detecting formation of transparent halo around the colony on LB-tributyrin agar plates. The amino acid sequence of BioHs was highly similar to the members of the BioH enzyme family involved in the biotin biosynthetic pathway; it showed the highest similarity (91%) with that of Serratia proteamaculans. To compare BioHs with other BioH enzymes, the relatively well-known bioHe gene of E. coli was cloned with PCR. After we achieved high-level expression of soluble BioHs and BioHe through the exploration of different culture conditions, the purified BioHs and BioHe enzymes were characterized in terms of specificity, activity, and stability. BioHe was generally more robust to a change in temperature and pH and an addition of organic solvents than BioHs. The two enzymes exhibited a strong preference for carboxylesterase rather than for thioesterase and were optimal at relatively low temperatures (20-40 degrees ) and alkaline pHs (7.5-9.0). The results in this study strongly suggested that both the BioHs and BioHe enzymes would be potential candidates for use as a carboxylesterase in many industrial applications. PMID- 19307764 TI - Kaempferol isolated from Nelumbo nucifera stamens negatively regulates FcepsilonRI expression in human basophilic KU812F cells. AB - Mast cells and basophils perform important functions as pivotal effector cells in IgE-mediated allergic reactions. KU812F cells, a human basophilic cell line isolated originally from chronic myelocytic leukemia, express a high affinity receptor of IgE, FcepsilonRI. Kaempferol was extracted and isolated from a methanolic extract of flavonoid-rich Nelumbo nucifera stamens. In the present study, the inhibitory effects of kaempferol on FcepsilonRI expression in human basophilic KU812F cells was examined. Flow cytometric analysis revealed that FcepsilonRI expression on the cell surface was suppressed in a concentration dependent manner when the cells were cultured with kaempferol. Moreover, RTPCR analysis showed that the mRNA levels for FcepsilonRI alpha- and gamma-chains were reduced as the result of kaempferol treatment in a concentration-dependent manner. Kaempferol showed its suppressive effects on intracellular Ca2+ concentration and histamine release from anti-FcepsilonRI alpha- chain antibody stimulated cells in a concentration-dependent manner. These observations indicate that kaempferol may exert antiallergic effect via downregulation of FcepsilonRI expression and degranulation. PMID- 19307765 TI - Hydrothermal acid pretreatment of Chlamydomonas reinhardtii biomass for ethanol production. AB - Certain microalgae have been known to use light and various carbon sources to produce carbohydrates, mainly in the form of starch. This is one of the pertinent feedstocks replacing agricultural products for the production of bioethanol by yeast. This study focuses upon dilute acid hydrothermal pretreatments at low cost and high efficiency to compete with current methods, and employs Chlamydomonas reinhardtii UTEX 90 as the feedstock. With dry cells of 5% (w/v), the algal biomass was pretreated with sulfuric acid (1-5%) under temperatures from 100 to 120oC, from 15 to 120 min. As a result, the glucose release from the biomass was maximum at 58% (w/w) after pretreatment with 3% sulfuric acid at 110 degrees for 30 min. This method enabled not only starch, but also the hydrolysis of other oligosaccharides in the algal cell in high efficiency. Arrheniustype of model equation enabled extrapolation of some yields of glucose beyond this range. The pretreated slurry was fermented by yeast, Saccharomyces cerevisiae S288C, resulting in an ethanol yield of 29.2% from algal biomass. This study suggests that the pretreated algal biomass is a suitable feedstock for ethanol production and can have a positive impact on large-scale applied systems. PMID- 19307766 TI - Optimization and scale-up of succinic acid production by Mannheimia succiniciproducens LPK7. AB - The effects of culture conditions on succinic acid production and its possible scale-up have been studied. Mannheimia succiniciproducens LPK7, engineered for enhanced production of succinic acid and reduced by-product secretion, was used for the experiments. Mannheimia succiniciproducens LPK7 is a knock-out strain of wild type deficient in the ldhA, pflB, and pta-ackA genes, and is derived from Mannheimia succiniciproducens MBEL55E. Process optimization of factors including optimal temperature, pH, carbon source, and nitrogen source was performed to enhance the production of succinic acid in flasks. To observe scale-up effects, batch fermentation was carried out at various working volumes. At a working volume of 7.0 l, the final succinic acid concentration and yield were 15.4 g/l and 0.86 g/g. This result shows similar amount of succinic acid obtained in lab scale fermentation, and it is possible to scale up to larger fermentors without major problems. PMID- 19307767 TI - Cloning of a gene encoding dextranase from Lipomyces starkeyi and its expression in Pichia pastoris. AB - A gene (lsd1) encoding dextranase from Lipomyces starkeyi KSM22 has been previously cloned, sequenced, and expressed in Saccharomyces cerevisiae. The gene consisting of 1,824 base pairs and encoding a protein of 608 amino acids was then cloned into and secretively expressed in Pichia pastoris under the control of the AOX1 promoter. The dextranase productivity of the P. pastoris transformant (pPIC9K-LSD1, 134,000 U/l) was approximately 4.2-fold higher than that of the S. cerevisiae transformant (pYLSD1, 32,000 U/l) cultured in an 8-l fermentor. Over 0.63 g/l of active dextranase was secreted into the medium after methanol induction. The dextranase of the P. pastoris transformant, as analyzed by SDS PAGE and Western blotting, showed only one homogeneous band. This dextranase of the P. pastoris transformant showed a broad band near 73 kDa. Rabbit monoclonal antibodies against a synthetic LSD1 peptide mix also recognized approximately 73 kDa. PMID- 19307768 TI - Screening and characterization of probiotic lactic acid bacteria isolated from Korean fermented foods. AB - To examine their potential as probiotics, acid and bile tolerance, antibiotics resistance, adhesion capacity to Caco- 2 and HT-29, and antibacterial activity, of LAB isolated from Korean fermented foods such as dongchimi, kimchi, Meju, and doenjang were assayed against foodborne pathogenic bacteria. DC 55, DC 136, DC 222, KC 117, KC 21, KC 24, KC 34, KC 43, MJ 301, MJ 54, SP 170, and SP 33 strains were resistant to acid and bile conditions. In particular, DC 55, DC 136, KC 24, KC 43, and MJ 301 strains were highly resistant to higher than 20 microg/ml concentrations of vancomycin, streptomycin sulfate, or amoxicillin, whereas, DC 222, KC 117, KC 21, KC 34, MJ 54, and SP 33 strains were susceptible to lower than 2microg/ ml concentrations of those antibiotics. The adhesion to HT-29 and Caco-2 cells varied with the strains tested in a strain-dependent manner. The highest level of adhesion was observed with DC 55, KC 21, KC 24, and MJ 301 strains, having higher than 50% of adhesion to HT-29 or Caco-2 cells. In addition, Staphylococcus aureus was the most sensitive to KC 21, showing an inhibition of about 70%, and the antibacterial activity of KC 21 against S. aureus resulted most likely from both organic acids and bacteriocin. Based on its phenotypic characteristics and utilization of various sugars, the KC 21 strain was identified as Lactobacillus plantarum. PMID- 19307769 TI - Identification of novel esterase from metagenomic library of Yangtze river. AB - A metagenomic library of surface-water microbes from the Yangtze River in China was constructed, and a novel esterase, designated as EstY, was isolated and characterized. EstY had 423 amino acids with an estimated molecular mass of 44 kDa and pI of 7.28. It hydrolyzed various pnitrophenyl esters (acetate, butyrate, caprate, caprylate, laurate, myristate, and palmitate) and its best substrate was p-nitrophenyl caprate (C8). The optimum pH for EstY activity was 9.0 and the optimum temperature was 50 degrees . Metal ions, such as Mn2+, Co2+, Hg2+, Zn2+, and Fe3+, strongly inhibited the activity of EstY, whereas Mg2+ was required for maximal activity. Activity remained in the presence of 10% alcohol, acetone, isopropanol, and dimethyl sulfoxide, respectively. An analysis of the amino acid sequence deduced from estY revealed that it had 7 closely related lipolytic enzymes. Moreover, a sequence analysis showed that EstY, like its 7 relatives, did not belong to any known lipolytic enzyme family. PMID- 19307770 TI - Expression of HpaGXooc protein in Bacillus subtilis and its biological functions. AB - HpaGXooc, from rice pathogenic bacterium Xanthomonas oryzae pv. oryzicola, is a member of the harpin group of proteins, eliciting hypersensitive cell death in non-host plants, inducing disease and insect resistance in plants, and enhancing plant growth. To express and secret the HpaGXooc protein in Bacillus subtilis, we constructed a recombinant expression vector pM43HF with stronger promoter P43 and signal peptide element nprB. The SDS-PAGE and Western blot analysis demonstrated the expression of the protein HpaGXooc in B. subtilis. The ELISA analysis determined the optimum condition for HpaGXooc expression in B. subtilis WBHF. The biological function analysis indicated that the protein HpaGXooc from B. subtilis WBHF elicits hypersensitive response (HR) and enhances the growth of tobacco. The results of RT-PCR analysis revealed that HpaGXooc induces expression of the pathogenesis-related genes PR-1a and PR-1b in plant defense response. PMID- 19307771 TI - Antibacterial and synergistic activity of isocryptomerin isolated from Selaginella tamariscina. AB - We investigated novel antibacterial and synergistic activities of isocryptomerin isolated from Selaginella tamariscina. Isocryptomerin showed potent antibacterial activity against Gram-positive and Gram-negative bacterial strains including clinical isolates of antibiotic-resistant species such as methicillin-resistant Staphylococcus aureus (MRSA). Additionally, we further investigated the synergistic activity of isocryptomerin with a conventional antibiotic against MRSA. The result indicated that isocryptomerin had considerable synergistic activity in combination with cefotaxime. In summary, the present study suggests that isocryptomerin may have potential as a novel therapeutic agent for treatment of infectious diseases by not only human pathogenic bacteria but also multidrug resistant bacteria. PMID- 19307772 TI - Differential expression of nuclear receptors in T helper cells. AB - Steroid hormones have long been known to have a profound influence on the immune system. Although the functions of these nuclear receptors in the development of T cells are fairly well studied, the differential expression of these receptors in T helper cells is poorly understood. Here, we investigated the differential expression of nuclear receptors and coregulators in Th1 and Th2 cells by genomewide microarray analysis. The result showed that several nuclear receptors and coregulators are differentially expressed in these cells. The result was confirmed by RT-PCR. The result showed that RXRalpha is highly expressed in Th2 cells. Overexpression of RXRalpha in a Jurkat human T cell line induced IL4 but not IFN-gamma gene expression, suggesting that RXRalpha plays a selective role in Th1 and Th2 differentiation. In summary, these results suggest that Th1/Th2 differentiation may be influenced by differential regulation of nuclear receptors and coregulators. PMID- 19307773 TI - Present situation of cedar pollinosis in Japan and its immune responses. AB - Recent observations have suggested significant worldwide increase in the prevalence of allergic rhinitis and cedar pollinosis. In Japan, Japanese cedar (Cryptometria japonica) and Japanese cypress (Chamaecyparis obtusa) pollens are considered to be the major unique allergens and their extent of dispersal is quite large, travelling more than 100km and thus causing serious pollinosis. Cedar pollinosis is a typical type 1 allergic disease by an adaptive immune response that occurs through the induction of allergen-specific effector T cells from naive T cells. We examined the number of Japanese cedar pollen specific memory Th cells in the peripheral blood of the patients and found that the cedar pollen specific IL-4-producing Th2 memory cells increased during the pollen season and decreased during the off-season. However, more than 60% of the cedar specific memory Th2 cells survived up to 8 months after the pollen season. Natural killer T(NKT) cells represent a unique lymphocyte subpopulation and their activity is not restricted to MHC antigens. NKT cells play an important role in innate immunity, however, the participation in development of allergic rhinitis could not be clarified. PMID- 19307774 TI - Peroxisome proliferator-activated receptor gamma negatively regulates allergic rhinitis in mice. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma(PPAR-gamma) has been shown to play an important role in the control of inflammatory responses acting on macrophages, mast cells, T cells, and eosinophils. The present study was aimed at investigating the effects of PPAR-gamma agonist on nasal symptoms and eosinophil accumulations in nasal mucosa by using a murine allergic rhinitis model. Furthermore, we examined the expression of PPAR-gamma in the nasal mucosa in mice. METHODS: BALB/c mice were sensitized and challenged intranasally with ovalbumin. Ciglitazone, a PPAR-gamma agonist, was administered orally 6 hours before each nasal challenge. RESULTS: Administration of PPAR-gamma agonist significantly decreased the number of nasal rubs, nasal histamine responsiveness, serum IgE, IL-5 production from the spleen, and eosinophilic infiltration in the nasal mucosa. Furthermore, PPAR-gamma was expressed in eosinophils and epithelial cells in the nasal mucosa by immunohistochemistry. CONCLUSIONS: PPAR-gamma was expressed in eosinophils and epithelial cells in the nasal mucosa. Also, the oral administration of ciglitazone is effective in upper airway allergic inflammation in mice. PMID- 19307775 TI - Allergic potency of Japanese cedar pollen Cry j 1 is reduced by a low concentration of hypochlorous acid generated by electolysis. AB - BACKGROUND: Although Japanese cedar (Cryptomeria japonica) pollinosis has developed into a health problem, few methods eradicate indoor allergens completely. In a recent study, however, the effectiveness of inactivation with sodium hypochlorite (NaOCl) treatment was revealed. Therefore, the present study aimed to elucidate the ability of chlorine bleach (NaOCl) to reduce the immunogenicity of the major allergenic protein of Japanese cedar (Cry j 1). METHODS: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, enzyme-linked immunosorbant assay, and skin testing were carried out in 7 individuals. RESULTS: The allergenic protein was undetectable using sodium dodecyl sulfate polyacrylamide gel electrophoresis and silver staining at a sodium hypochlorite/allergenic protein molar ratio of 457. Western blotting with human sera showed the same dose-dependent efficacy. The immunogenicity of the purified protein and cedar pollen was also demonstrated on enzyme-linked immunosorbant assay to be reduced by sodium hypochlorite treatment in a dose- and time-dependent manner. Moreover, sodium hypochlorite-treatment inhibited the skin test response to the protein in all 7 individuals. CONCLUSIONS: Hypochlorous acid generated by electrolysis is an effective method for significantly reducing the immunogenicity of Cry j 1. PMID- 19307776 TI - Allergy to Salsola Kali in a Salsola incanescens-rich area: role of extensive cross allergenicity. AB - BACKGROUND: Pollens from the Salsola spp. are an important source of respiratory allergy in tropical countries. Our aim was to characterize the IgE binding proteins of S. incanescens pollen extract and study its cross-reactivity with S. kali pollen allergens. METHODS: Prick tests with S. kali and S. incanescens pollen extracts were performed on eight respiratory allergy patients from Mashhad, Northeast Iran. The antigenic profiles and IgE-binding patterns of S. kali and S. incanescens pollen extracts were compared by SDS-PAGE and Western blotting, using individual sera from the salsola pollen-sensitive patients. Cross reactivity of proteins in the two weeds was assessed by IgE- immunoblotting inhibition. RESULTS: S. kali and S. incanescens pollen extracts showed similar IgE-binding profiles in Western blotting. The IgE binding components of 39, 45, 66 and 85 kDa were detected in both pollen extracts. Furthermore, inhibition of the immunoblots revealed extensive inhibition of IgE binding to proteins and a close relationship between these two weeds allergens. CONCLUSIONS: S. incanescens pollen is a potent allergen source with several IgE binding components that shows a close allergenic relationship with S. kali. Our results suggest that in S. incanescens-rich areas, S. kali pollen extracts could be used as a diagnostic reagent for allergic patients to S. incanescens pollen. PMID- 19307777 TI - Enzyme-linked immunosorbent assays with high sensitivity for antigen-specific and total murine IgE: a useful tool for the study of allergies in mouse models. AB - BACKGROUND: In studies on allergies in mouse models, IgE production is an essential parameter to be evaluated. Here, we examine the effect of commercially available immunoreaction enhancer solutions and different blocking reagents in enzyme-linked immunosorbent assay (ELISA) for total or antigen-specific murine IgE in order to improve the assays. METHODS: Sera from mice immunized with recombinant house dust mite major allergens, Der f 1 and Der p 1, were used for the assays. Total IgE was measured by sandwich ELISA using monoclonal antibodies against murine IgE. Antigen-specific IgE was assayed using allergen-coated plates. Sensitivity or signal intensity in ELISA was compared among conditions differing in the use of enhancer solutions, blocking reagents, or monoclonal antibodies, and incubation time. RESULTS: Use of enhancer solutions improved the sensitivity of ELISA for total IgE by approximately 30-fold of that using a conventional buffer. A blocking reagent caused more unwanted enhancement of the background signal in blank wells in ELISA for total IgE compared with another blocking reagent, however, improved signal intensity in ELISA for antigen specific ELISA without significant enhancement of the background signal. Optimal assay conditions were determined. CONCLUSIONS: Enhancer solutions are effective in improving ELISAs for total and antigen-specific murine IgE. Selection of blocking reagents was important to decrease unwanted enhancement of background signals and was effective in enhancing signals for positive samples. The ELISAs improved in this study are useful for the study of allergies in mouse models. PMID- 19307778 TI - Recognition of T cell epitopes unique to Cha o 2, the major allergen in Japanese cypress pollen, in allergic patients cross-reactive to Japanese cedar and Japanese cypress pollen. AB - BACKGROUND: Pollens from species of the Cupressaceae family are one of the most important causes of respiratory allergies worldwide. Many patients with pollinosis have specific IgE to both allergens from Japanese cedar and Japanese cypress pollen. We set out to identify T cell epitopes in Cha o 2, the second major allergen of Japanese cypress pollen. METHODS: T cell lines (TCL) and T cell clones (TCC) specific to Cha o 2 were generated from allergic patients cross reactive to Japanese cedar and Japanese cypress pollen. T cell epitopes in Cha o 2 were identified by responses of TCL stimulated with overlapping peptides. Abilities of IL-4/IFN-gamma production by TCC were evaluated using enzyme immunoassay. RESULTS: Using TCL, 11 dominant and subdominant T cell epitopes were identified in Cha o 2. The subsets of TCC were predominantly of T helper 2-type. A T cell epitope p141-160 in Cha o 2 and corresponding peptide in Cry j 2 showed high homology. Although TCC PC.205.159 responded to stimulation with p141-160 in Cha o 2, it did not respond with corresponding peptide in Cry j 2, therefore, the T cell epitope was unique to Cha o 2. CONCLUSIONS: Eleven T cell epitopes that were identified are unique to Cha o 2. Cha o 2 is a putative aeroallergen that can potentially sensitize human T cells. We concluded that generation of T cells specific to Cha o 2 in allergic patients acts as one of the causes of continuous allergic symptoms in April. PMID- 19307779 TI - Acute generalized exanthematous pustulosis (AGEP) triggered by a spider bite. AB - BACKGROUND: Acute generalized exanthematous pustulosis (AGEP) is a rare and severe cutaneous reaction usually triggered by drugs. Other causative factors such as viral infections are rarely involved. In this study, we report a case of AGEP caused by a spider bite. CASE SUMMARY: A 56-year-old woman was referred to the allergy unit after a spider bite at the left popliteal fossa, while gardening, 5 days earlier. The offending spider was captured and identified by an entomologist as belonging to the Loxosceles rufescens species. No acute reaction was observed; however, after 24 hours, due to the occurrence of typical dermonecrotic skin lesions associated with erythema and edema, Cefuroxime and Clindamycin were administered intramuscularly after medical advice was given. Almost 72 hours after the spider bite, an erythematous and partly edematous eruption appeared locally in the gluteus area bilaterally, which progressively expanded to the trunk, arms and femors. Within 24 hours dozens of small, pinhead sized, non-follicular pustules were present, mainly in the folds. The patient complained of a burning sensation of the skin in addition to pruritus; and simultaneously had a fever of 38-39 degrees C as the eruption expanded. DISCUSSION: A spider bite may represent a possible causative factor of AGEP. A spider's venom contains sphingomyelinase that stimulates the release of IL8 and GM-CSF, which are involved in AGEP pathogenesis. Whether or not the con-current use of antibiotics has an effect in AGEP appearance when combined with a spider's venom, cannot be excluded. PMID- 19307780 TI - What clinical alternatives to whole liver transplantation? Current status of artificial devices and hepatocyte transplantation. AB - Shortage of organ donors limits the number of possible liver transplantations. Alternative therapies for treatment of liver failure are currently being developed: (i) extracorporeal artificial liver devices; (ii) bioartificial liver devices using hepatocytes; and (iii) hepatocyte transplantation. The objective of these strategies is to bridge patients with liver failure until a suitable liver allograft is obtained for transplantation or the patient's own liver regenerates sufficiently to resume normal function. In this review, we discuss these strategies and summarize the current status of clinical experience. PMID- 19307781 TI - Increased frequency and responsiveness of PSA-specific T cells after allogeneic hematopoetic stem-cell transplantation. AB - BACKGROUND: Therapies for localized prostate cancer include curative surgery and radiotherapy while treatment of metastatic disease is often inefficient. Graft versus-tumor effects of allogeneic stem-cell transplantation (ASCT) have been described for several types of solid tumors but have not been reported for prostate cancer. We, therefore, investigated the potential of ASCT as treatment for noncurable prostate cancer. METHODS: A patient underwent ASCT from his human leukocyte antigen (HLA)-identical sister as treatment for his metastatic prostate adenocarcinoma. Frequencies of prostate-specific T cells in the peripheral blood of the patient, ASCT donor and a group of control individuals were determined by flow cytometry using pentameric HLA-A2 complexes containing peptides derived from the prostate-specific antigen (PSA). Cytotoxic activity of PSA-peptide-specific T cells against peptide-pulsed target cells was analyzed ex vivo by Cr-release assays. RESULTS: Stable clinical and laboratory remission lasting more than 4 years was observed after ASCT. Using HLA-containing pentamers with PSA-derived peptides we could detect prostate-specific CD8+ T cells in this patient at high frequencies over several months. Furthermore, higher frequencies of PSA-specific T cells were revealed in the blood of the patient and female controls when compared with healthy males. CONCLUSIONS: Lymphocytes from the peripheral blood of the recipient, but not from donor or other tested control individuals, exhibited ex vivo cytotoxic activity against target cells pulsed with the relevant synthetic peptides and efficiently expanded in vitro following specific restimulations. Thus, the results of this study indicate that female to male ASCT can increase the frequency and enhance specific-responsiveness of PSA-specific T cells in transplant recipients. PMID- 19307782 TI - Binding of thymoglobulin to natural killer cells leads to cell activation and interferon-gamma production. AB - Thymoglobulin is an antithymocyte globulin preparation used in hematopoietic stem cell transplantation (HSCT) to prevent rejection and graft-versus-host disease. Because natural killer (NK)-cell alloreactivity improves HSCT outcome, but only in patients receiving thymoglobulin, we investigated the in vitro effects of thymoglobulin on purified NK cells. Thymoglobulin binding to NK cells and NK-cell activation were assessed by flow cytometry. NK surface targets for thymoglobulin were determined by competition inhibition assays using monoclonal antibodies. Chromium 51 (Cr) release assay, Annexin V combined with 7-amino-actinomycin D staining, and carboxyfluorescein diacetate succinimidyl ester staining were used to study cytotoxic activity, apoptosis/cell death, and NK-cell proliferation, respectively. Interferon (IFN)-gamma production was determined by ELISA. Thymoglobulin, thymoglobulin derived-F(ab')2 fragments as well as rabbit IgG bound NK cells, and competed strongly with anti-CD16. Thymoglobulin enhanced the expression of activation (CD69 and NKG2D) and degranulation (CD107a) markers on NK cells. It competed with CD18 binding and decreased NK activity, but not interleukin-15-induced killer activity. Effects on apoptosis/cell death and proliferation were minimal. F(ab')2 fragments and rabbit IgG strongly induced IFN gamma production by NK cells. Thymoglobulin binds to NK cells by CD16 by its variable and constant regions. The decrease in NK-cell cytotoxic activity is restored by interleukin-15, and contrasts sharply with the induction of activation, degranulation, and IFN-gamma production. These data support the hypothesis that thymoglobulin treatment is required to observe the improvement in HSCT outcome by NK-cell alloreactivity. PMID- 19307783 TI - Local expression of B7-H4 by recombinant adenovirus transduction in mouse islets prolongs allograft survival. AB - BACKGROUND: Allogeneic pancreatic islet transplantation has the potential to cure type 1 diabetes. One of the barriers to islet transplantation is the alloreactive T-cell response between donors and recipients. Costimulatory molecules, which play a major role in the regulation of the immune response to antigens during graft rejection, may be used to inhibit allograft destruction. B7-H4 is one such member in the costimulatory family, which has established negative regulatory function of T-cell responses. METHODS: To determine whether local expression of B7-H4 protein can protect beta cells from damage in islet allotransplantation, we have constructed a recombinant adenovirus expressing a B7-H4 complementary deoxyribonucleic acid (Ad-B7-H4). To study the in vivo effects of B7-H4 expression on islet graft survival, adenovirus-transduced islets from donor Balb/c mice were transplanted into streptozotocin-diabetic C57BL/6 mice (n=12). RESULTS: Expression of B7-H4 in islets by Ad-B7-H4 transduction at an optimized condition did not inhibit glucose-stimulated insulin secretion of the treated islets. The recipient mice transplanted with Ad-B7-H4-transduced islets established euglycemia for a longer time (mean 56.5 days), compared with control mice transplanted with Ad-LacZ-transduced islets (mean 14.5 days, [n=12, P<0.001]). Splenocytes isolated from the recipients of Ad-B7-H4-transduced islets showed hyporesponsiveness to alloantigenic stimulation, compared with control recipients. CD45 and insulin staining of the graft transplanted with Ad-B7-H4 transduced islets indicated the preservation of beta cells and decrease of infiltrating immune cells. CONCLUSIONS: Local expression of B7-H4 prolongs islet allograft survival in vivo, suggesting translational potential for beta-cell replacement with reduced immune injury. PMID- 19307784 TI - Human leukocyte transmigration across Galalpha(1,3)Gal-negative porcine endothelium is regulated by human CD18 and CD99. AB - BACKGROUND: In pig-to-human xenotransplantation cross-species receptor interactions mediate cellular infiltration and rejection of porcine grafts. However, the mechanisms responsible for recruitment of human leukocyte subsets across porcine endothelial cells (EC) remain largely unknown. Here, we investigated the role of CD99, CD18, and Galalpha(1,3)Gal (Gal) in this process. METHODS: Adhesion and transmigration of human peripheral blood mononuclear cell (PBMC) subsets on Gal and Gal porcine EC (pEC) and on human EC was analyzed using a two-compartment system separated by a permeable membrane. The mechanisms of human PBMC recruitment to pEC were investigated by blocking cell surface receptors and by differentially measuring adhesion and transendothelial migration (TEM). RESULTS: Blocking of CD18, but not CD99, decreased human PBMC adhesion on pEC, whereas blocking of CD18 or CD99 strongly reduced the subsequent human PBMC TEM across pEC. The inhibitory effect of CD99 blockade was slightly stronger across pEC as compared with human EC. A critical role for Gal in TEM of human monocytes, B, natural killer (NK), NK/T, and T cells was excluded by evaluating TEM across pEC derived from Gal and Gal pigs. CONCLUSIONS: CD99 and CD18, but not Gal, play a critical role in human monocyte and lymphocyte TEM across pEC, and their respective porcine ligands may serve as targets to specifically inhibit human leukocyte recruitment in pig-to-human xenotransplantation. PMID- 19307785 TI - Evidence for Induced Expression of HLA Class II on Human Islets: Possible Mechanism for HLA Sensitization in Transplant Recipients. AB - Recent reports have shown that islet transplant recipients develop antibodies against donor human leukocyte antigen (HLA) class I and II. Because human islets do not express HLA class II under normal conditions, mechanisms underlying induction of the anti-class II response are unclear. We hypothesized that under inflammatory conditions, islets will have induced expression of HLA class II leading to sensitization. Isolated human islets were divided into two groups. Group 1 was cultured at 37 degrees C as control; group 2 was cultured similarly in presence of tumor necrosis factor alpha and interferon gamma. After treatment, islets were analyzed for expression of HLA class II using real-time polymerase chain reaction, immunofluorescence and flow cytometry. Furthermore, serum from an islet transplant recipient who developed anti-class II antibody was tested by flow cytometry for immunoglobulin (Ig) binding to cytokine-stimulated islets. Real-time polymerase chain reaction analysis for gene transcripts of class II transactivator, HLA-DRagr;, and HLA-DRbeta1 showed maximum 9.38-, 18.95-, and 46.5-fold increase, respectively in group 2 when compared with control at 24 hr. Cytokine treatment increased HLA class II expression markedly on both alpha and beta cells in islets as evidenced by fluorescent imaging and flow cytometric analysis. When patient serum was analyzed by flow cytometry, both IgM and IgG binding was observed in cytokine-treated, HLA class II matched islet cells alone. We conclude that inflammation leads to induced expression of HLA class II on transplanted islet cells potentially causing antidonor sensitization and adversely impacting islet transplant outcomes. PMID- 19307786 TI - Deficiency of protein kinase C-theta facilitates tolerance induction. AB - BACKGROUND: Protein kinase C-theta (PKCtheta) mediates critical T-cell receptor signals required for T-cell activation. We have recently shown that PKCtheta knockout (PKCtheta, H-2b) T cells, when transferred into T/B cell-deficient mice, failed to reject fully allogeneic (H-2d) cardiac grafts and that transgenic expression of antiapoptotic Bcl-xL gene in PKCtheta T cells restored allograft rejection. METHODS: We used PKCtheta mice as recipients of cardiac allografts, compared with wild-type (WT) cardiac allograft transplantation. Anti-CD154 monoclonal antibody (MR1) and human CTLA4Ig were sued to induce donor-specific tolerance. T-cell proliferation, T-cell subsests, nuclear factor kappa B (NF kappaB) activation, and Bax and Bcl-xL were analyzed. RESULTS: Although suboptimal anti-CD154 monoclonal antibody or human CTLA4Ig failed to delay cardiac allograft rejection in WT mice, the same therapy induced long-term survival of cardiac allografts in PKCtheta mice. Donor-type second cardiac allografts (H-2d) were accepted, and third-party heart allografts (H-2k) were rejected by tolerant PKCtheta mice. However, tolerance state could not be effectively transferred with T cells from tolerance PKCtheta mice. Compared with WT mice, reduced NF-kappaB activation, T-cell proliferation, and T-cell infiltration in PKCtheta spleens were observed. PKCtheta mice reveal reduced CD4/CD25/FoxP3, Th1/Th17 subsets, and mouse MHC class II (IE)-reactive CD4Vbeta11 T cells. Apoptotic molecule, Bax, was increased and antiapoptotic molecule, Bcl xL, was reduced in PKCtheta spleen cells. CONCLUSION: We concluded that PKCtheta mice have a defected alloimmune response and are susceptible to tolerance induction, which is associated with a clonal deletion of T-cell subsets. PMID- 19307787 TI - Endogenous sulfur dioxide aggravates myocardial injury in isolated rat heart with ischemia and reperfusion. AB - BACKGROUND: Ischemia-reperfusion (I/R) injury is an important clinical problem. This article investigated the role of sulfur dioxide (SO2) in the regulation of cardiac function and in the pathogenesis of cardiac I/R injury in isolated rat heart. METHODS: Rat hearts isolated on a Langendorff apparatus were divided into control, I/R, I/R+SO2, and I/R+hydroxamate groups. Hydroxamate is an inhibitor of SO2 synthetase. I/R treatment was ischemia for 2 hr in hypothermic solution (4 degrees C), then reperfusion/rewarming (37 degrees C) for 60 min. Cardiac function was monitored by MacLab analog to a digital converter. Determination of sulfite content involved reverse-phase high performance liquid chromatography with fluorescence detection. Myoglobin content of coronary perfusate was determined at 410 nm. Myocardial malondialdehyde (MDA) was determined by thiobarbituric acid method, and conjugated diene (CD) was extracted by chloroform. 5,50-Dithiobis-2-nitrobenzoic acid was used to determine glutathione (GSH). RESULTS: The results showed that I/R treatment obviously increased myocardial sulfite content, and sulfite content of myocardium was negatively correlated with the recovery rate of left-ventricle developed pressure and positively correlated with the leakage of myoglobin. In postreperfusion, myocardial function recovery was decreased by SO2. During reperfusion, myocardium released enzymes, MDA and CD level were increased but myocardial GSH content was depressed with the treatment of SO2 donor. Incubation of myocardial tissue with SO2 significantly increased MDA and CD generation. CONCLUSIONS: Endogenous SO2 might be involved in the pathogenesis of myocardial I/R injury, and its mechanism might be associated with an increase in lipid peroxide level and a decrease in GSH generation. PMID- 19307788 TI - Development of BK nephropathy in recipients of simultaneous pancreas-kidney transplantation. AB - BACKGROUND: BK nephropathy (BKVN) is a significant cause of graft dysfunction in kidney transplant recipients, but its course in simultaneous pancreas-kidney (SPK) recipients is less well studied. The presence of dual organs limits the ability to reduce maintenance immunosuppression, typically the first intervention in the management of BKVN. METHODS: A single center, retrospective review was conducted of 205 SPK transplants performed from January 1, 2000 to April 30, 2006. RESULTS: The 5-year actuarial cumulative rate of BKVN was 5.6%. Diagnosis occurred at a median of 20 months after transplant; mean serum creatinine was 2.6, and geometric mean BK serum viral load was 709,274 copies/mL at diagnosis. There was no statistical difference in the cumulative rate according to the use of induction therapy: rabbit antilymphocyte globulin (5-year rate 6.8%, 4/59), alemtuzumab (5-year rate 5.1%, 5/146). Treatment consisted of immunosuppression reduction and half received cidofovir. Eight of nine kidney allografts eventually failed, but all patients retained pancreatic allograft function. CONCLUSIONS: BKVN occurs in 5.6% of SPK recipients. There is no difference in the cumulative rate of BKVN between patients who received alemtuzumab or rabbit antilymphocyte globulin. PMID- 19307789 TI - Impact of des-gamma-carboxy prothrombin and tumor size on the recurrence of hepatocellular carcinoma after living donor liver transplantation. AB - BACKGROUND: Because many patients who did not meet the Milan criteria have survived long after undergoing living donor liver transplantation (LDLT), extended criteria for recipient with hepatocellular carcinoma (HCC) are therefore considered to be necessary. METHODS AND RESULTS: A total of 90 consecutive adult LDLT recipients with HCC between 1996 and 2007 were reviewed. The recurrence-free survival rates of all 90 patients were 86.0%, 81.3%, and 81.3% at 1, 3, and 5 years, respectively. Fourteen of 90 patients developed a recurrence of tumor after the LDLT. The tumor recurrences were diagnosed within 1 year after the LDLT in 11 (78.6%) patients. In a multivariate analysis, both the tumor size of less than 5 cm (P=0.0202) and the des-gamma-carboxy prothrombin (DCP) level of less than 300 mAU/mL (P=0.0001) were found to be favorable independent factors for the recurrence of HCC after LDLT. Therefore, the authors devised new selection criteria for HCC patients (a tumor size of <5 cm or a DCP of <300 mAU/mL). The 1 , 3-, and 5-year overall or recurrence-free survival rates of the 85 patients who met the new criteria were 92.3%, 85.9%, and 82.7%, or 90.5%, 87.0%, and 87.0%, respectively, which were significantly different from those of the five patients who did not meet the new criteria (P<0.0001). CONCLUSIONS: A combination of two factors, namely the tumor size and the DCP level, was found to be useful for expanding the selection of LDLT candidates for HCC. PMID- 19307790 TI - Progression of renal dysfunction in cardiac transplantation after the introduction of everolimus in the immunosuppressive regime. AB - The aim of this study was to analyze, in heart transplant patients, if renal function improvement after cyclosporine replacement by everolimus persists at the middle term and its predictors. We studied prospectively 56 patients in whom conversion was consecutively made. Forty-five patients completed the follow-up period. Significant improvement was observed at 6 and 12 months in plasma creatinine levels (1.92+/-0.7 vs. 1.67+/-0.6 and 1.69+/-0.6 mg/dL; P=0.047) and glomerular filtration rate (43.9+/-17 vs. 52.5+/-23 and 51.3+/-22.3 mL/min; P=0.004). Glomerular filtration rate increased in 32 patients (71%). Baseline characteristics comparison showed a lower percentage of patients with smoking history and new onset diabetes in responders group, but only previous smoking was shown as independent factor (Exp B: 0.083; 95% confidence interval: 0.010-0.793; P=0.024). No differences regarding age, gender, body mass index, disease leading to transplantation, time between transplantation and replacement, cardiovascular risk factors, lipid levels, and hematologic parameters were found. PMID- 19307791 TI - Combined en-bloc liver-pancreas transplantation in patients with liver cirrhosis and insulin-dependent type 2 diabetes mellitus. AB - We report about our experience with combined en-bloc liver-pancreas transplantation in 14 patients with liver cirrhosis and insulin dependent type 2 diabetes mellitus. Exocrine drainage was achieved by duodeno-duodenostomy. Median posttransplant follow-up is currently 92.5 months. All patients were rendered independent from insulin therapy shortly after transplantation. Levels of glycosylated hemoglobin normalized in all recipients. Mean fasting C-peptide values increased from pretransplant 7.0+/-1.7 ng/mL to 10.5+/-2.9 ng/mL 3 months posttransplantation (P<0.001). One recipient (7.1%) developed recurrent exogenous insulin dependence 7 years after transplantation. Pancreas allograft rejection was confirmed by endoscopic biopsy of donor duodenum mucosa in two patients (14.3%). Calculated 5- and 7-year survival is currently at 64.3% and 64.3%, respectively. Our results indicate that combined en-bloc liver-pancreas transplantation using duodeno-duodenostomy is technically feasible and leads to excellent long-term control of glucose metabolism in patients with liver cirrhosis and insulin-dependent type 2 diabetes. PMID- 19307792 TI - No occurrence of de novo HLA antibodies in patients with early corticosteroid withdrawal in a 5-year prospective randomized study. AB - The purpose of this study was to determine the effect of early corticosteroid cessation on the occurrence of de novo human leukocyte antigen (HLA) antibody posttransplant. Renal transplant recipients (n=37) were randomized to early corticosteroid withdrawal at day 7 posttransplant (n=21 patients), or to chronic steroids (n=16), all in combination with thymoglobulin as induction agent, tacrolimus and mycophenolic acid as maintenance therapy. To establish the time course of HLA antibody appearance, sera collected pretransplant and for up to 5 years posttransplant were screened for the appearance of HLA antibodies. In this 5-year longitudinal study, only one patient in the control group developed a de novo donor-specific HLA antibody. We conclude that renal transplant recipients on steroid withdrawal by the end of week 1 are not at higher risk for developing HLA antibodies compared with a standard steroid regimen up to 5 years posttransplant. PMID- 19307793 TI - Multicenter evaluation of a novel endothelial cell crossmatch test in kidney transplantation. AB - BACKGROUND: Despite their clinical importance, clinical routine tests to detect anti-endothelial cell antibodies (AECA) in organ transplantation have not been readily available. This multicenter prospective kidney transplantation trial evaluates the efficacy of a novel endothelial cell crossmatch (ECXM) test to detect donor-reactive AECA associated with kidney allograft rejection. METHODS: Pretransplant serum samples from 147 patients were tested for AECA by a novel flow cytometric crossmatch technique (XM-ONE) using peripheral blood endothelial progenitor cells as targets. Patient enrolment was based on acceptance for transplantation determined by donor lymphocyte crossmatch results. RESULTS: Donor reactive AECA were found in 35 of 147 (24%) patients. A significantly higher proportion of patients with a positive ECXM had rejections (16 of 35, 46%) during the follow-up of at least 3 months compared with those without AECA (13 of 112, 12%; P<0.00005). Both IgG and IgM AECAs were associated with graft rejections. Mean serum creatinine levels were significantly higher in patients with a positive ECXM test at 3 and 6 months posttransplant. CONCLUSIONS: XM-ONE is quick, easy to perform on whole blood samples and identifies patients at risk for rejection and reduced graft function not identified by conventional lymphocyte crossmatches. PMID- 19307794 TI - Organ procurement and transplantation network/united network for organ sharing histocompatibility committee collaborative study to evaluate prediction of crossmatch results in highly sensitized patients. AB - BACKGROUND: The requirement for a prospective crossmatch limits some organ allocation to local areas. The delay necessitated by the crossmatch restricts the distance across which offers can be made without unduly increasing the ischemia time. A collaborative study involving 14 transplant centers was undertaken by the Organ Procurement and Transplantation Network/United Network for Organ Sharing (OPTN/UNOS) Histocompatibility Committee to evaluate the accuracy with which the detection of unacceptable human leukocyte antigen (HLA) antigens by most advanced solid phase immunoassays can predict crossmatch results. In addition, using actual patients' unacceptable HLA antigens, the number of compatible donors that would have been available from the OPTN deceased kidney donors during 2002 to 2004 were investigated. METHODS: Panel reactive antibodies were performed by conventional or solid phase assays, and crossmatches were performed by cytotoxicity or flow cytometry. Analyses were stratified for T and B cell and by method of identifying unacceptable HLA antigens and crossmatch techniques. RESULTS: Combination of solid phase immunoassays and flow cytometry crossmatches resulted in a higher prediction rates of positive T cell (86.1%-93.5%) and B-cell crossmatches (91%-97.8%). Prediction of negative crossmatches based on different combination of panel reactive antibodies and crossmatch techniques varied from 14.3% to 57.1%. Furthermore, numerous potential compatible donors were identified for each patient, regardless of their ethnicity, in the OPTN database, when predicted incompatible ones were excluded. CONCLUSIONS: The above results showed that with the advent of solid phase immunoassays, HLA antibodies can now be accurately detected resulting in prediction of crossmatch outcome. This should facilitate organ allocation and prevents shipment of organs to distant incompatible recipients. PMID- 19307795 TI - Donor-directed HLA antibodies before and after transplantectomy detected by the luminex single antigen assay. AB - BACKGROUND: Donor-directed antibodies (DDA) have been shown to result in poor graft survival. This study was designed to analyze antibody appearance and patient and graft characteristics related to antibody formation in patients who lost their graft at different time points after transplantation. METHODS: Pre- and posttransplant sera of 56 DDA-negative first transplant patients were screened for human leukocyte antigen (HLA) class I and II DDA by the Luminex single antigen assay (LSA). All patients were treated with calcineurine inhibitor based immunosuppression. RESULTS: Three of 56 patients proved DDA positive by LSA before transplantation. Eighty-one percent of the remaining 53 patients became DDA class I or II positive or both; 16% before and 84% after transplantectomy. Class I antibodies were produced in 84% and class II in 77% of the recipients. Based on time of transplantectomy, three groups were created as follows: less than or equal to 1 month, 1 to 6 months, and more than 6 months. The groups proved to be significantly different for HLA class II mismatch and acute rejection. All recipients in group 1 to 6 months proved to be DDA positive. Logistic regression analysis showed that DDA positivity for class I was related to higher donor age and donor type (nonheart beating), class II to higher donor age and class II mismatch. CONCLUSIONS: Donor-directed HLA antibodies after transplantation were demonstrated in 81% of first transplant recipients, all of whom were DDA negative by LSA before transplantation. The majority of the antibodies was found after transplantectomy. These findings may have to be taken into consideration in the allocation of organs of marginal donors such as older or nonheart beating kidneys. PMID- 19307796 TI - Probabilistic modeling of cytomegalovirus infection under consensus clinical management guidelines. AB - BACKGROUND: Cytomegalovirus (CMV) is the most common viral pathogen after renal transplantation and remains a major therapeutic challenge with important clinical and economic implications from both direct and indirect consequences of infection. METHODS: This 5-year study modeled the relationship between CMV infection and biopsy-proven graft rejection, graft loss, or death after renal transplantation in an inception cohort using Canadian consensus guidelines for CMV management as a component of a detailed cost-analysis of viral infection. RESULTS: Probabilities of CMV viremia and syndrome/disease among 270 sequential graft recipients were 0.27 and 0.09, respectively; 91% of cases occurred in the first 6 months. Probability of CMV infection as the first event was 0.29, with a probability of subsequent biopsy-proven acute rejection (BPAR) of 0.05 (mean: 62+/-26 days, range: 32-85 days), whereas the probability of BPAR as the first event was 0.18, with a probability of subsequent CMV infection of 0.38 (mean: 63+/-31, range: 27-119 days). Probability of freedom from both CMV infection and BPAR throughout the period of observation was 0.53. Time-dependent Cox analysis showed that neither donor/recipient CMV risk stratum nor CMV infection influenced the risks of BPAR (P=0.24; P=0.74) or of graft loss or death (P=0.26; P=0.34). In contrast, BPAR significantly increased the risk of both subsequent CMV infection (hazard ratio=1.77, P=0.03) and of graft loss or death (hazard ratio=8.31, P<0.0001). CONCLUSIONS: Although current antiviral therapy seems to mitigate the reported deleterious effects of CMV infection on BPAR or graft survival, BPAR remains a significantly risk factor for both CMV infection and functional graft survival. PMID- 19307797 TI - Identification of the optimal donor quality scoring system and measure of early renal function in kidney transplantation. AB - BACKGROUND: The early identification of kidney allografts at risk of later dysfunction has implications for clinical practice. Donor quality scoring systems (preoperative) and measures of early allograft function (first week postoperative) have previously shown practical utility. This study aimed to determine the optimal parameter(s) (preoperative and postoperative) with greatest predictive power for the development of subsequent allograft dysfunction. METHODS: Consecutive deceased donor renal transplants (n=217) were studied. In each, the following measures were assessed: Preoperative donor quality scores: expanded criteria donor status; Deceased Donor Score (Nyberg et al., Am J Transplant 2003;3:715); Donor Risk Score (Schold et al., Am J Transplant 2005; 5(4 pt 1): 757); and delayed graft function (DGF) Nomogram (Irish et al., J Am Soc Nephrol 2003; 14: 2967). Postoperative early function measures: dialysis requirement and duration; extended DGF definition (Boom et al., Kidney Int 2000; 58: 859); creatinine at day 5 and day 7; creatinine reduction ratios at day 2 and day 7; and urine output posttransplantation. Primary outcome measures were creatinine at 12 months and the development of chronic kidney disease stage 4T. RESULTS: Of donor scoring systems, Donor Risk Score was best associated with subsequent allograft function. Of early function measures: the extended definition of DGF, creatinine at day 5, and dialysis duration showed greatest predictive power in the patient population overall, those not requiring postoperative dialysis, and those requiring dialysis, respectively. No scores or early function measures were associated with change in creatinine between 6 and 12 months. CONCLUSIONS: This study validates and identifies the optimal early predictive parameter available for kidney transplant recipients, with implications for refining early postoperative management and potential utility in organ allocation policy. PMID- 19307798 TI - Dual-source CT coronary angiogram in heart transplant recipients in comparison with dobutamine stress echocardiography for detection of cardiac allograft vasculopathy. AB - Conventional coronary angiography (CCA) is the gold standard in the diagnosis of cardiac allograft vasculopathy (CAV) in heart transplant recipients. Dobutamine stress echocardiography (DSE) is a useful technique for screening. Dual-source computed tomography (DSCT) is the last generation of computed tomography scanners, which could be useful to noninvasively assess CAV. Thirty cardiac transplant recipients underwent DSE and DSCT coronary angiogram. Exclusion criteria were as follows: renal insufficiency, iodinated contrast media allergy, less than 12 months since transplant, and unstable clinical conditions. DSE showed ischemia in two patients. At DSCT scan 13 patients had a normal angiogram, 13 ones wall thickening and four significant diseases. DSCT showed a sensitivity of 100% with a specificity of 92%. DSCT allowed detection of more patients with CAV than DSE. Four patients showed significant CAV at DSCT compared with two at DSE. Thirteen patients showed initial signs of disease at DSCT despite a normal DSE. PMID- 19307799 TI - Calcineurin inhibitor sparing with mycophenolate in kidney transplantation: a systematic review and meta-analysis. AB - BACKGROUND: Limiting the exposure of kidney transplant recipients to calcineurin inhibitors (CNIs) has potential merit, but there is no clear consensus on the utility of current strategies. In an attempt to aid clarification, we conducted a systematic review and meta-analysis of randomized trials that assessed CNI sparing (minimization or elimination) with mycophenolate as sole adjunctive immunosuppression. METHODS: The search strategy identified trials where CNI sparing was accompanied by the continuation of, or conversion to, mycophenolate and compared with standard or higher dose CNI therapy. Two investigators independently examined each trial for eligibility, quality, and outcome measures. Additional subgroup analyses were assessed: (1) de novo CNI sparing; (2) elective CNI sparing beyond 2 months posttransplantation; and (3) CNI sparing for transplant dysfunction. RESULTS: Nineteen randomized controlled trials met the inclusion criteria permitting analysis of 3312 renal transplant recipients with median follow-up of 12 months. CNI sparing significantly improved glomerular filtration rate (weighted mean difference 4.4 mL/min, 95% confidence interval [CI] 2.9-5.9, P<0.001); with some evidence, albeit weak, of improved graft survival (odds ratio 0.72, 95% CI 0.52-1.01, P=0.06). Acute rejection rates were only increased after elective CNI elimination (odds ratio 2.23, 95% CI 1.57-3.17, P<0.001). There were no significant differences in mortality, malignancy or incidence of infections. CONCLUSIONS: CNI sparing strategies with adjunctive mycophenolate may play an important role in kidney transplant recipients. Improvements in short-term graft function, and possibly graft survival, are achievable. Longer term studies are needed to substantiate the short-term benefits, and refining elective CNI elimination protocols may help to reduce the risk of rejection. PMID- 19307800 TI - Requirement of protocol biopsy before and after complete cessation of immunosuppression after liver transplantation. AB - BACKGROUND: Operational tolerance is defined as long-term acceptance of a transplanted organ after complete cessation of immunosuppression (IS), but may not always protect against antigen-dependent changes in graft morphology. METHOD: IS free patients after living-donor liver transplantation (LDLT) underwent protocol biopsy (tolerance group [Gr-Tol]) and were evaluated for rejection and fibrosis. The degree of fibrosis was compared with those in the patients on maintenance IS group (Gr-IS) and the base line normal liver group (Gr-BS). When bridging fibrosis or progression of fibrosis was observed, IS was reintroduced or increased in Gr-Tol or in the patients in the weaning process. RESULTS: Neither acute nor chronic rejection was observed. The degree of fibrosis, however, was significantly greater in Gr-Tol than those in Gr-IS and Gr-BS. In Gr-Tol, the number of graft infiltrating FOXP3 cells was significantly increased, the interval between LDLT and biopsy plus the donor age was significantly longer, and recipient age at LDLT was significantly younger, compared with those in Gr-IS. However, none of these three parameters correlated with the degree of fibrosis. In 7 of 11 patients in whom IS was reintroduced or increased, the improvement of fibrosis was observed by the subsequent biopsy. CONCLUSION: Grafts of operationally tolerant patients after LDLT did not exhibit acute or chronic rejection, but they exhibited fibrosis. It remains elusive whether fibrosis observed in tolerant grafts is antigen dependent. The finding that after [corrected] the reintroduction or the increase of IS fibrosis was improved supported the possibility that fibrosis in operationally tolerant patients was antigen dependent. PMID- 19307801 TI - Thrombosis of developmental venous anomalies of the brain after liver transplantation. PMID- 19307802 TI - Pediatric brain death diagnosis in the view of organ donation in France. PMID- 19307803 TI - High-dose chemotherapy followed by autologous stem-cell transplantation in liver transplanted multiple myeloma patients: a report of two cases. PMID- 19307804 TI - Acute phosphate nephropathy in a kidney transplant recipient with delayed graft function. PMID- 19307805 TI - A critical appraisal of "transfusion strategies for patients in pediatric intensive care units" by Lacroix J, Hebert PC, Hutchison, et al (N Engl J Med 2007; 356:1609-1619). AB - OBJECTIVE: To review the findings and discuss the implications of transfusion strategies in stable critically ill children. DESIGN: A critical appraisal of the article "Transfusion strategies for patients in pediatric intensive care units" by Lacroix J, Hebert PC, Hutchison, et al, published in the N Engl J Med in 2007 with literature review. FINDINGS: In this prospective, randomized, controlled, noninferiority trial the authors compared a liberal transfusion strategy, using a transfusion threshold of 9 g/dL, to a conservative transfusion strategy, using a transfusion threshold of 7 g/dL. The primary end point was multiple organ dysfunction syndrome (MODS) or progression of MODS. The authors found that when comparing the restrictive transfusion strategy to the liberal strategy, the absolute risk reduction for developing new or progressive MODS was only 0.4% (95% confidence interval, -4.6 -5.5). Using the restrictive protocol, the number needed to treat to prevent one red blood cell (RBC) transfusion was only two. The number of RBC units per patient in the restrictive group was 0.9, and in the liberal group was 1.7 (p < 0.001). When comparing the two strategies there was a relative reduction of 96% in the number of patients who had any transfusion exposure and a relative decrease of 44% in the number of transfusions administered in the restrictive strategy. CONCLUSIONS: Using a restrictive transfusion protocol with a transfusion threshold of 7 g/dL in stable critically ill children is as safe as using a liberal protocol and can decrease the number of patients exposed to RBC transfusions. PMID- 19307806 TI - Reduction of hospital mortality and of preventable cardiac arrest and death on introduction of a pediatric medical emergency team. AB - OBJECTIVE: To determine the effect of a medical emergency team (MET) on the incidence of unexpected cardiac arrest and death. DESIGN: Comparison of retrospective data (pre-MET) before introduction of MET with prospective data after introduction of MET system (post-MET). SETTING: Tertiary care pediatric hospital. PATIENTS: A total of 104,780 admissions during a 41-month period pre MET; 138,424 admissions during 48 months post-MET. INTERVENTIONS: Introduction of a MET. RESULTS: Total hospital deaths decreased from 4.38 to 2.87/ 1000 admissions (risk ratio 0.65, 95% confidence interval [CI] 0.57-0.75, p < 0.0001). Ward unexpected death decreased from 13 (0.12/1000) to 6 (0.04/1000) (risk ratio 0.35, 95% CI 0.13- 0.92, p = 0.03) but unexpected cardiac arrests did not change from 0.19/1000 to 0.17/1000 (risk ratio 0.91, 95% CI 0.50 -1.64, p = 0.75). Thirty-four hospital deaths, including three unexpected deaths (1 out of 72 MET calls), were prevented each year of MET operation. Preventable cardiac arrest (children whose symptoms or signs fulfilled MET calling criteria) decreased from 17 (0.16/ 1000) to 10 (0.07/1000) (risk ratio 0.45, 95% CI 0.20-0.97, p = 0.04) and in whom death decreased from 12 to 2 (0.11/1000 to 0.01/1000) (risk ratio 0.13, 95% CI 0.03-0.56, p = 0.001). Nonpreventable cardiac arrest (children whose symptoms or signs did not fulfill MET calling criteria) increased from 3 to 14 (0.03/1000 to 0.10/1000, p = 0.03) but death did not increase. Survival from cardiac arrest increased from 7 of 20 patients to 17 of 23 (risk ratio 2.11, 95% CI 1.11- 4.02, p = 0.01). Annual calls for urgent assistance were 202 in the post MET era and 46 during the pre-MET era (ratio 4.4:1). CONCLUSIONS: Introduction of a MET was associated with reduction of total hospital death and reduction of preventable cardiac arrest and death with increased survival in wards of a pediatric hospital. MET calling criteria identified some but not all children at risk of unexpected cardiac arrest and death. PMID- 19307807 TI - Hyperglycemia and insulin therapy in the critically ill child. AB - OBJECTIVE: To canvass the current opinion of pediatric intensivists in the United Kingdom regarding the importance of hyperglycemia,their approach to management, and their views in relation to a potential intervention trial of tight glycemic control. DESIGN: Electronic survey comprising a 17-point questionnaire,along with six clinical scenarios describing cases of bronchiolitis, septic shock, major trauma, postcardiac surgery, necrotizing enterocolitis,and acute respiratory distress syndrome. SETTING: All pediatric intensive care units in the United Kingdom(n = 21). PARTICIPANTS: All consultant pediatric intensivists in the United Kingdom (n = 117). INTERVENTIONS: None. MEASUREMENTS AND METHODS: The response rate was 75% (88 of 117), representing all 21 units. Although all respondents administer insulin to hyperglycemic patients, only one in two believe hyperglycemia causes significant morbidity in critically ill children. Predominant factors influencing the decision to start insulin are severity of hyperglycemia (95% of responders), duration of hyperglycemia (85%), and patient diagnosis (49%). The blood glucose threshold for starting insulin varied widely from 6.1 to>15 mmol x L(-1) (110-270 mg x dL(-1)) for each of the six scenarios,with poor agreement between intensivists within each center(intraclass correlation coefficient of 0.29). Sixty-seven percent of respondents, representing 14 of the 21 units reported having no written policy or guideline in their unit for management of hyper-glycemia in nondiabetic children. Eighty-six percent (76) of intensivists would be prepared to participate in a trial of "tight glycemic control" but only 63% (55) would be prepared to include all patient groups. Only 20% would be willing to target blood glucose below 6.1 mmol x L(-1) (110 mg x dL(-1)) in the tight control group. CONCLUSIONS: This survey suggests significant variation in the management of hyperglycemia across the UK Practice varies even among intensivists from the same unit, reflecting the fact that few units have 'an agreed written guidance in place.' The majority of intensivists would be prepared to participate in a trial of tight glycemic control. PMID- 19307808 TI - Using acupuncture for acute pain in hospitalized children. AB - OBJECTIVE: Clinical study to determine the acceptability and feasibility of acupuncture for acute postoperative pain control in hospitalized children. DESIGN: Nonrandomized clinical trial. SETTING: A single, tertiary referral pediatric intensive care unit. PATIENTS: A total of 20 patients aged 7 months to 18 years. Eleven of the patients had posterior spinal fusion surgery and the remaining nine patients had other surgical diagnoses. INTERVENTIONS: Two 10- to 15-minute sessions of acupuncture 24-48 hours apart. OUTCOME MEASURES AND RESULTS: The treatment was highly accepted (27 patients were approached and 4 patients refused; of the 23 patients enrolled, 20 patients completed the study). Acupuncture was well tolerated by patients without adverse events related to treatment. In follow-up interviews, 70% of both parents and patients believed acupuncture helped the child's pain. Eighty-five percent of the parents said they would pay out of pocket for acupuncture if not covered by insurance. The pain scores, vital signs, and narcotic usage were recorded before and several times after acupuncture. In posterior spinal fusion patients, the mean pain scores (0 10) immediately before and 4 and 24 hours after acupuncture were: 3.7, 1.7, and 3.1, respectively, after the first acupuncture session and 3.7, 2.2, and 3.1, respectively, after the second session. In the other surgical cohort, the mean pain scores immediately before and 4 and 24 hours after the first session of acupuncture were 2.5, 0.3, and 1.6, respectively. CONCLUSIONS: Our results support that acupuncture is highly accepted and feasible in critically ill, postoperative pediatric patients with acute pain. Our findings suggest that acupuncture may be a potentially useful adjunctive tool for acute pediatric postoperative pain management. A randomized, controlled clinical trial is warranted to confirm these findings. PMID- 19307809 TI - Blood product transfusions and clinical outcomes in pediatric patients with acute lung injury. AB - OBJECTIVE: There are data suggesting that blood product transfusions increase the risk of developing acute lung injury (ALI) in adults, and may be associated with increased mortality in adults with ALI. A possible association between transfusions and adverse outcomes of pediatric patients with ALI has not been studied previously. We tested the hypothesis that blood product transfusions to pediatric patients with ALI within the first 72 hours of the diagnosis would be associated with increased mortality and prolonged mechanical ventilation. DESIGN: An epidemiologic database of pediatric ALI prospectively gathered from July 1996 to May 2000 was analyzed. SETTING: Children were enrolled from both a tertiary referral hospital and a major community children's hospital. PATIENTS: Three hundred fifteen patients who met the 1994 American European Consensus Committee definition of ALI between the ages of 36 weeks corrected gestational age and 18 years. MAIN OUTCOME MEASURE: Mortality in the pediatric intensive care unit. RESULTS: Multivariate analyses indicated that the transfusion of fresh-frozen plasma (FFP) was associated with increased mortality, independent of the severity of hypoxemia (Pao2/Fio2), presence of multiple organ system failure or disseminated intravascular coagulation (odds ratio = 1.08, 95% confidence interval = 1.00-1.17, p = 0.04). FFP transfusion was analyzed as a continuous variable, so that for each milliliter of FFP transfused per kilogram patient body weight per day, the odds of death increased by 1.08. There was a trend toward an association of the transfusion of FFP with a fewer number of days of unassisted ventilation (regression coefficient = -0.21, 95% confidence interval = -0.42 0.01, p = 0.06). CONCLUSIONS: The transfusion of FFP is associated with an increased risk of mortality in children with ALI. The association between FFP and mortality in children with ALI should be investigated further. PMID- 19307810 TI - Cardiopulmonary interaction. AB - OBJECTIVE: To highlight and review the physiology and pathophysiology of cardiopulmonary interaction in the critically ill pediatric patient. DATA SOURCE: A MEDLINE-based literature source. OUTLINE OF REVIEW: This review is divided into two sections: 1) The physiologic basis of cardiopulmonary interaction, and 2) critical clinical conditions in which cardiac and/or pulmonary dysfunction impact each other in the provision of adequate oxygen delivery. The physiology section focuses on the original research identifying fundamental volume-pressure and pressure-flow relationships and then proceeds to discuss how changes in intrathoracic pressure and lung volume affect ventricular loading conditions. The clinical section chooses several common scenarios in which this normal physiology is altered and an understanding of the impact of these physiologic aberrations on cardiac and/or pulmonary function is required to make sound management decisions. CONCLUSIONS: Improving oxygen transport balance is the primary goal in the management of children with life-threatening disorders. To optimize the opportunity for a successful outcome, the intensivist must understand the complex relationship between the cardiac and pulmonary systems in the effort to provide sufficient oxygen to meet the body's metabolic demands. Furthermore, the application of therapies separately designed to support the function of the heart and the function of the lungs may be synergistic or antagonistic, further complicating the management scheme. We conclude that this review will encourage the reader to pursue further literature or perhaps engage in further research related to this often underappreciated but vital interplay between cardiac and pulmonary functions. PMID- 19307811 TI - Severe invasive community-associated methicillin-resistant Staphylococcus aureus infections in previously healthy children. AB - OBJECTIVE: An increase in community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections has been reported in the literature. Most severe, life-threatening infections were previously thought to be associated with chronically ill or frail patients. Our pediatric intensive care unit (PICU) has seen a recent dramatic increase in primary, severe invasive CA-MRSA infections in healthy children. DESIGN/SETTING: A retrospective chart review of all previously healthy patients admitted to our 19-bed combined medical-surgical PICU with a primary diagnosis of severe invasive, culture-proven CA-MRSA disease during the past 6 years. RESULTS: Eleven previously healthy patients were admitted to our PICU with severe, primary, invasive CA-MRSA infections from March 2006 through September 2007, in contrast to no patients meeting these criteria in the preceding 5 years. The mortality rate was 27%, compared with an overall PICU mortality rate during the study period of <7%. The mean PICU length of stay of these patients was 14.9 days, compared with an average PICU length of stay of 2.4 days. Despite initiation of treatment with vancomycin at admission to the PICU in all but one case, patients took a mean of 5.7 days to convert to negative blood cultures. Eight patients had bacteremia longer than 4 days. Six of the patients developed bilateral necrotizing pneumonia requiring prolonged mechanical ventilation. CONCLUSIONS: Severe CA-MRSA infections in healthy children are increasing at an alarming rate in our institution. This acute rise in incidence, coupled with an alarmingly high associated mortality rate, raises important questions about the initial empirical antibiotic therapy we use in caring for patients presenting with suspected life-threatening CA-MRSA disease. Vancomycin monotherapy may not be adequate treatment for severe CA-MRSA infections. PMID- 19307812 TI - Effect of alternative chest compression techniques in infant and child on rescuer performance. AB - OBJECTIVE: Current chest compression (CC) guidelines for an infant recommend a two-finger (TF) technique with lone rescuer and a two- thumb (TT) technique with two rescuers, and for a child either an one hand (OH) or a two hand (TH) technique with one or two rescuers. The effect of a 30:2 compression:ventilation ratio using these techniques on CC quality and rescuer fatigue is unknown. We hypothesized that during lone rescuer CC, TT technique, in infant and TH in child achieve better compression depth (CD) without additional rescuer fatigue compared with TF and OH, respectively. DESIGN: Randomized observational study. SETTING: University-affiliated pediatric hospital. SUBJECTS: Adult healthcare providers certified in basic life support or pediatric advanced life support. INTERVENTIONS: Laerdal baby advanced life support trainer and Resusci junior manikin were modified to digitally record CD, compression pressure (CP) and compression rate. Sixteen subjects were randomized to each of the four techniques to perform 5 minutes of lone rescuer 30:2 compression:ventilation cardiopulmonary resuscitation. Rescuer heart rate (HR) and respiratory rate were recorded continuously and the recovery time interval for HR/respiratory rate to return to baseline was determined. Subjects were blinded to data recording. Groups were compared using two-sample, two-sided Student's t tests. MEASUREMENTS AND MAIN RESULTS: Two-thumb technique generated significantly higher CD and peak CP compared with TF (p < 0.001); there was no significant difference between OH vs. TH. TF showed decay of CD and CP over time compared with TT. Compression rate (per minute) and actual compressions delivered were not significantly different between groups. No significant differences in fatigue and recovery time were observed, except the TT group had greater increase in the rescuer's HR (bpm) from baseline compared with TF group (p = 0.04). CONCLUSIONS: Two-thumb compression provides higher CD and CP compared with TF without any evidence of decay in quality and additional rescuer fatigue over 5 minutes. There was no significant difference in child CC quality or rescuer fatigue between OH and TH. Two-thumb technique is preferred for infant CC and our data support the current guidelines for child CC. PMID- 19307814 TI - Neuron-specific enolase and S-100B are associated with neurologic outcome after pediatric cardiac arrest. AB - OBJECTIVE: To characterize the pattern of serum biochemical markers of central nervous system injury (neuron-specific enolase [NSE], S-100B, plasminogen activator inhibitor-1 [PAI-1]) after pediatric cardiac arrest and determine whether there is an association between biomarker concentrations and neurologic outcome. DESIGN: Prospective, observational study. SETTING: Urban, tertiary care children's hospital. PATIENTS: Cardiac arrest survivors, n = 35. INTERVENTIONS: Serial blood sampling, pediatric cerebral performance category, and standardized neurologic examination. MEASUREMENTS AND MAIN RESULTS: Serial serum NSE and S 100B concentrations over 96 hrs and PAI-1 at 24 hrs were measured in children (age <18 yrs) who had return of spontaneous circulation following cardiac arrest. Neurologic outcome was prospectively categorized as poor if the change in pre- to postarrest pediatric cerebral performance category was > or =2. Biomarker concentrations were compared between outcome groups and between survival groups using longitudinal analysis correcting for multiple comparisons. Median levels (25th, 75th percentiles) are reported. Receiver operating characteristic analyses were performed at all time points. Biomarker concentrations showed statistically significant differences. Of the 35 patients, neurologic outcomes were poor in 19, with 15 deaths. Median NSE concentrations differed by outcome when measured at > or =48 hrs, and by survival at > or =24 hrs. S-100B concentrations were not significantly associated with neurologic outcome. S-100B levels were associated with survival outcome at > or =48 hrs. PAI-1 levels were not significantly associated with either neurologic or survival outcomes. CONCLUSIONS: The timing, intensity, and duration of serum NSE and S-100B biomarker concentration patterns are associated with neurologic and survival outcomes following pediatric cardiac arrest. Serum NSE concentrations at > or =48 hrs are associated with neurologic outcome, whereas serum S-100B levels at > or =48 hrs are associated with survival. Prospective analysis of these markers may help to predict outcomes and guide postresuscitative therapies. PMID- 19307815 TI - Is "rescue" therapy ethical in randomized controlled trials? AB - OBJECTIVE: There is a commonly held belief that randomized, placebo-controlled trials in pediatric critical care should incorporate "rescue" therapy (open-label administration of active drug) when a child's condition is deteriorating. The ethical, conceptual, and analytic challenges related to rescue therapy in randomized trials can be misrepresented. DESIGN: Narrative review. METHODS: The ethical basis of rescue therapy, the equipoise concept, and intention-to-treat analysis are examined in the setting of a hypothetical randomized trial comparing corticosteroids vs. placebo in pediatric septic shock. FINDINGS: The perceived need for rescue therapy may be partly motivated by the moral imperative to save a child's life. However, allowing rescue therapy in a trial is misconceived and inconsistent with equipoise regarding the efficacy of the study drug. If rescue therapy is permitted, intention-to-treat analysis can only compare immediate vs. delayed use of the study drug. When rescue therapy is beneficial, the observed treatment effect is substantially diminished from true effect of the study drug, leading to increased sample size and thereby placing more children at risk (18 "excess" placebo-arm deaths occur in our hypothetical example). Analysis of a trial incorporating rescue therapy cannot definitively assess overall efficacy of the agent, or distinguish beneficial or harmful treatment effects related to timing of drug use. CONCLUSIONS: Although a rescue therapy component in a randomized trial may be perceived as ethically desirable, inconsistency of rescue therapy with full equipoise may itself raise significant ethical concerns. Increased sample sizes expose more children to the risks of study participation, including death. Researchers should be aware that clinical trials designed with rescue therapy cannot definitively determine the beneficial or harmful effects of a treatment per se, and can only assess the effects of delayed vs. immediate provision of the treatment. PMID- 19307816 TI - Mechanical component failures in 28,171 neonatal and pediatric extracorporeal membrane oxygenation courses from 1987 to 2006. AB - OBJECTIVE: To provide a descriptive summary of mechanical component failure associated with extracorporeal membrane oxygenation (ECMO), and to examine patient and ECMO variables that may be associated with mechanical component failure and guide further study. We hypothesized that duration of ECMO, era of ECMO, indication for ECMO, age of patient, and center ECMO volume would be associated with mechanical component failure. DESIGN: Retrospective cohort study. PATIENTS AND METHODS: Extracorporeal Life Support Organization registry was queried for all neonatal and pediatric ECMO courses recorded. Each ECMO course was treated as an independent event, and was included if duration was > or =25 hrs with occurrence between 1987 and 2007. Courses with a duration >458 hrs or with an indication for ECMO during cardiopulmonary resuscitation were excluded from analysis. Mechanical component failure data were extracted from the Extracorporeal Life Support Organization registry for the oxygenator, raceway, other tubing, pigtail connectors, heat exchanger, and air in the circuit. RESULTS: A total of 28,171 independent ECMO courses were included for analysis, of which 14.9% were associated with a mechanical component failure. Duration of ECMO, age group of patient, era of ECMO, and indication for ECMO were all associated with mechanical component failure. From our predictive model, we observed a continuous nonlinear relation suggesting increasing probability of mechanical component failure with increasing duration of ECMO support. CONCLUSIONS: Mechanical component failure over the course of this study was infrequent during neonatal and pediatric ECMO, and declined across eras as experience with the therapy grew. Increasing duration of ECMO was associated with an increasing probability of mechanical component failure. Indication for ECMO and patient age were also statistically associated with mechanical component failure probability, but ECMO center volume was not. PMID- 19307818 TI - Risk factors for central line-associated bloodstream infection in a pediatric cardiac intensive care unit. AB - OBJECTIVE: To identify risk factors for central line-associated bloodstream infection (BSI) in patients receiving care in a pediatric cardiac intensive care unit. DESIGN: Matched case-control study. SETTING: CICU at Children's Hospital Boston. PATIENTS: Central line-associated BSI cases were identified between April 2004 and December 2006. We identified two randomly selected control patients who had a central vascular catheter and were admitted within 7 days of each index case. MEASUREMENTS AND MAIN RESULTS: Univariate and multivariate conditional logistic regression analyses were used to identify risk factors for central line associated BSI. In a secondary analysis, risk factors for central line-associated BSI in those cases who underwent cardiac surgery were sought. During the study period, 67 central line-associated BSIs occurred in 61 patients. Independent risk factors for central line-associated BSI were nonelective admission for medical management (odds ratio [OR] = 6.51 [1.58-26.78]), the presence of noncardiac comorbidities (OR = 4.95 [1.49-16.49]), initial absolute neutrophil count <5000 cells/uL (OR = 6.17 [1.39-27.48]), blood product exposure > or =3 units (OR = 5.56 [1.35-22.87]), central line days > or =7 (OR = 6.06 [1.65-21.83]), and use of hydrocortisone (OR = 28.94 [2.55-330.37]). In those patients who underwent cardiac surgery (n = 37 cases and 108 controls), independent risk factors for central line-associated BSI were admission weight < or =5 kg (OR = 3.13 [1.01 9.68]), Pediatric Risk of Mortality III score > or =15 (OR = 3.44 [1.19-9.92]), blood product exposure > or =3 units (OR = 3.38 [1.28-11.76]), and mechanical ventilation for > or =7 days (OR = 4.06 [1.33-12.40]). CONCLUSIONS: Unscheduled medical admissions, presence of noncardiac comorbidities, extended device utilization, and specific medical therapies are independent risk factors for central line-associated BSI in patients receiving care in a pediatric cardiac intensive care unit. PMID- 19307817 TI - Management of severely injured children in road accidents in France: impact of the acute care organization on the outcome. AB - OBJECTIVE: To examine the impact of acute care management on outcome in children severely injured in road accidents. DESIGN AND SETTING: Prospective follow-up study conducted in 12 French pediatric intensive care units over a 24-month period. PATIENTS: Excluding those in refractory shock or in brain death at admission, a total of 125 children aged <17 years admitted to the pediatric intensive care unit with severe trauma (Injury Severity Score > or =16) were included. RESULTS: Intracranial pressure (ICP) monitoring and admission into a trauma resuscitation room (TRR) were used as proxy markers for the center management aggressiveness. Centers which admitted to TRR and monitored ICP when indicated in >75% of cases were called aggressive centers. Children with an ICP monitoring indication admitted to a TRR and monitored, as well as those without an indication treated in a TRR, were judged appropriately managed. A poor outcome at pediatric intensive care unit discharge was defined as a difference between the baseline and discharge pediatric overall performance category above 3, or a hospital death. Children with traumatic brain injury appropriately managed in a less-aggressive center were more likely to have a poor outcome than those appropriately managed in an aggressive center (odds ratio 7.56, 95% confidence interval 1.5-38.4), after adjustment for severity, age, and type of road user. CONCLUSIONS: The management in a more aggressive center for children admitted to TRR and monitored for ICP, when indicated, is associated with a better outcome. This could be explained by a more extensive experience in trauma management. PMID- 19307819 TI - Enteral feeding algorithm for infants with hypoplastic left heart syndrome poststage I palliation. AB - BACKGROUND: Infants with hypoplastic left heart syndrome (HLHS) experience a high incidence of growth failure in the postoperative period following stage I palliation. Because of an increased risk of necrotizing enterocolitis in this population, clinicians may be reluctant to initiate early enteral feedings. Published guidelines for initiating and advancing enteral feedings in this population are limited. OBJECTIVE: To test the safety and efficacy of an enteral feeding algorithm in infants with HLHS following stage I palliation. DESIGN: Single-center, prospective case series with historical comparisons. SETTING: Pediatric cardiovascular intensive care unit in tertiary care children's hospital. PATIENTS: The study group consisted of consecutive patients > or =35 wks gestational age and weight > or =2 kg admitted to our cardiac intensive care unit over an 18-month period following stage I palliation of HLHS (n = 36). Excluded were nonsurvivors, patients supported on extracorporeal membrane oxygenation or those with a history of NEC or fetal intervention. These data were compared with a similar cohort of patients admitted to the cardiac intensive care unit over an 18-month period before the implementation of the feeding algorithm (n = 27). INTERVENTION: A feeding algorithm was implemented in the study group in the postoperative period for initiation and advancement of enteral nutrition. MEASUREMENTS AND MAIN RESULTS: The median duration of total parenteral nutrition was significantly higher in the control group (116 vs. 51 hrs; p = 0.03) compared with the study group. The median time to achieve recommended daily allowance of calories defined as 108 kcal/kg per day was significantly reduced in the study group (9 vs. 13 days; p = 0.01). Despite the rapid advancement of enteral feedings on the algorithm, there was no incidence of NEC in the study group compared with 11% in the control group. CONCLUSION: The use of an enteral feeding algorithm is a safe and effective means of initiating and advancing enteral nutrition in infants with HLHS following stage I palliation. PMID- 19307820 TI - Inflammatory and oxidative parameters in cord blood as diagnostic of early-onset neonatal sepsis: a case-control study. AB - OBJECTIVE: To determine whether levels of interleukin (IL)-6, IL-10, and oxidative parameters in umbilical cord blood could contribute as an indicator of neonatal sepsis in recognized high-risk neonates. DESIGN: Prospective, case control study. SETTING: Neonatal intensive care unit. SUBJECTS: One hundred twenty consecutive preterm neonates who had at least one other risk factor for early-onset neonatal sepsis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Umbilical cord blood samples were obtained for the determination of IL-6, IL-10, thiobarbituric acid reactive substances (TBARS), and protein carbonyls levels. Neonates were divided prospectively in two groups: control and septic. All parameters were higher in septic patients compared with control (IL-6 184.6 +/- 72.7 vs. 58.9 +/- 19.1, p < 0.01; IL-10 171.4 +/- 59.2 vs. 79.9 +/- 17.9, p < 0.01; TBARS 10.1 +/- 2.8 vs. 4.2 +/- 2.5, p < 0.01; protein carbonyls 2.4 +/- 1.2 vs. 1.15 +/- 0.5, p < 0.01, respectively, septic vs. control). In addition, these parameters were higher in the subgroup of culture-positive septic patients compared with control. IL-6 and TBARS had equivalent areas under the receiver operator characteristic (ROC) curve (0.88); IL-10 (0.80) and protein carbonyls (0.73) had lower areas. Multivariate logistic regression comparing IL-6 and TBARS in terms of the relative risk for neonatal sepsis demonstrated that TBARS was a better predictor, being independently associated with neonatal sepsis. CONCLUSION: Our findings demonstrated that cord blood IL-6, IL-10, and oxidative stress markers were significantly higher in infants with neonatal sepsis, and only TBARS levels were independently related to the development of neonatal sepsis in our sample. PMID- 19307821 TI - Pediatric organ donation: a national survey examining consent rates and characteristics of donor hospitals. AB - OBJECTIVE: To characterize the eligible pediatric donor pool in the United States by age, consent rate, location, and cause of donor loss. STUDY DESIGN: Survey. SETTING: Organ procurement organizations in the United States. PATIENTS: Pediatric patients who suffered brain death in the United States in 2005 and who were medically suitable for organ donation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We determined the number of patients, age < or =17 yrs, who were eligible for organ donation and the consent rate by age. Each hospital in which donation occurred was characterized by the presence of a pediatric intensive care unit (PICU), a pediatric critical care medicine (PCCM) fellowship, solid organ transplant programs, and level I trauma programs. Additional information was obtained on the number of donation after cardiac death donors and eligible donors lost due to medical examiner refusals and deterioration before organ recovery. The number of reported eligible pediatric donors in 2005 was 1330. The consent rate was 69.2% with higher consent rates in eligible donors 12 yrs of age and older. Eligible donors were spread across a large number of hospitals with few having ten or more eligible donors. Variability exists among hospitals in consent rate and number of donors per 100 PICU beds. The presence of a level I trauma program and/or a PCCM fellowship was associated with higher numbers of donors per 100 PICU beds. Ninety-four eligible donors were lost before recovery of organs due to medical examiner denials or cardiac arrest. Donation after cardiac death accounted for 37 donors. CONCLUSIONS: Overall pediatric consent rates were 69.2% but varied by age. Eligible donors were found most often in hospitals with level I trauma programs or PCCM fellowship programs. Few hospitals had >10 eligible donors in a 12-month period. This study is the first to describe in detail the U.S. pediatric donor population. PMID- 19307855 TI - Psychiatric aspects of nonpsychiatric disorders in older adults. PMID- 19307856 TI - Is family history of depression a risk factor for poststroke depression? Meta analysis. AB - OBJECTIVE: To determine whether family history of psychiatric disorder constitutes a risk factor for the development of poststroke depression. DESIGN: A meta-analysis setting: patients examined for depression following stroke seen in acute care, rehabilitation hospital, or outpatient care settings. PARTICIPANTS: All patients who were reported in the world's literature in English language publications in which information was provided about the existence or not of poststroke depression and the presence or absence of a family history of psychiatric disorder. MEASUREMENTS: The frequency of family history of psychiatric disorder was determined for each study as well as the relationship of family history to the presence of poststroke depression. RESULTS: Based on data obtained from 903 patients with stroke, the fixed model analysis found a risk ratio of 1.51 and the random model a risk ratio of 1.46 for the existence of poststroke depression if there is a positive family history of psychiatric disorder compared with a negative family history. CONCLUSIONS: The existence of a positive family history of psychiatric disorder constitutes a risk factor for development of poststroke depression. The role of family history in poststroke depression, however, appears to be substantially lower than among elderly depressed patients without evidence of vascular disease. PMID- 19307857 TI - Aging, neurocognition, and medication adherence in HIV infection. AB - OBJECTIVE: To evaluate the hypothesis that poor adherence to highly active antiretroviral treatment (HAART) would be more strongly related to cognitive impairment among older than among younger HIV-seropositive adults. SETTING AND PARTICIPANTS: A volunteer sample of 431 HIV-infected adult patients prescribed self-administered HAART was recruited from community agencies and university affiliated infectious disease clinics in the Los Angeles area. MEASUREMENTS: Neurocognitive measures included tests of attention, information processing speed, learning/memory, verbal fluency, motor functioning, and executive functioning. Medication adherence was measured using microchip-embedded pill bottle caps (Medication Event Monitoring System) and self-report. Latent/structural analysis techniques were used to evaluate factor models of cognition and adherence. RESULTS: Mean adherence rates were higher among older (>or=50 years) than younger (<50 years) HIV-positive adults. However, latent/structural modeling demonstrated that neurocognitive impairment was associated with poorer medication adherence among older participants only. When cognitive subdomains were examined individually, executive functioning, motor functioning, and processing speed were most strongly related to adherence in this age group. CD4 count and drug problems were also more strongly associated with adherence among older than younger adults. CONCLUSIONS: Older HIV-positive individuals with neurocognitive impairment or drug problems are at increased risk of suboptimal adherence to medication. Likewise, older adults may be especially vulnerable to immunological and neurocognitive dysfunction under conditions of suboptimal HAART adherence. These findings highlight the importance of optimizing medication adherence rates and evaluating neurocognition in the growing population of older HIV-infected patients. PMID- 19307858 TI - Engagement in persons with dementia: the concept and its measurement. AB - PURPOSE: The aim of this article is to delineate the underlying premises of the concept of engagement in persons with dementia and present a new theoretical framework of engagement. SETTING/SUBJECTS: The sample included 193 residents of seven Maryland nursing homes. All participants had a diagnosis of dementia. METHODOLOGY: The authors describe a model of factors that affect engagement of persons with dementia. Moreover, the authors present the psychometric qualities of an assessment designed to capture the dimensions of engagement (Observational Measurement of Engagement). Finally, the authors detail plans for future research and data analyses that are currently underway. DISCUSSION: This article lays the foundation for a new theoretical framework concerning the mechanisms of interactions between persons with cognitive impairment and environmental stimuli. Additionally, the study examines what factors are associated with interest and negative and positive feelings in engagement. PMID- 19307859 TI - Patterns of mild cognitive impairment after treatment of depression in the elderly. AB - OBJECTIVES: Late-life depression (LLD) is associated with persistent cognitive impairment in a subset of individuals. The purpose of this study was to 1) examine the frequency and characteristics of cognitive diagnoses (Mild Cognitive Impairment [MCI], dementia) among remitted elderly depressed subjects and 2) to compare the prevalence rate and correlates of cognitive diagnoses with those of comparison subjects. DESIGN: Crosssectional. SETTING: Outpatient geriatric mental health clinic. PARTICIPANTS: The authors examined cognitive diagnoses among 109 subjects age 65 and older, after depression treatment response and 65 never depressed, age- and education-equated comparison subjects. MEASUREMENTS: Cognitive diagnoses were independently established by the University of Pittsburgh's Alzheimer's Disease Research Center. Bivariate and multivariate analyses were conducted to examine the role of specific risk factors for cognitive diagnosis among depressed subjects. RESULTS: Relative to comparison subjects, nearly twice as many depressed subjects were diagnosed with MCI or dementia (48% versus 28%). Of the 109 depressed subjects, 38% were diagnosed with MCI (63% amnestic, 37% nonamnestic). The majority of amnestic MCI subjects (85%) had the multiple domain subtype. Age, but not age of onset or lifetime depression duration, predicted cognitive diagnosis. CONCLUSIONS: Despite adequate depression treatment response, 48% of remitted depressed subjects had a cognitive diagnosis. Of the 38% diagnosed with MCI, there was high representation among both the amnestic and the nonamnestic subtypes, suggesting heterogeneity in cognitive course and outcomes in LLD. PMID- 19307861 TI - The Patient Health Questionnaire 8: current depressive symptoms among U.S. older adults, 2006 Behavioral Risk Factor Surveillance System. AB - OBJECTIVES: To examine the prevalence and sociodemographic predictors of current depressive symptoms among adults aged 65 years and older. DESIGN AND SETTING: Participants were obtained from the 2006 Behavioral Risk Factor Surveillance System, a population-based list-assisted random-digit-dialed telephone survey of the noninstitutionalized U.S. adults. PARTICIPANTS: A total of 45,534 participants aged 65 years and older were interviewed. MEASUREMENTS: Participants completed the Patient Health Questionnaire 8. Those with a Patient Health Questionnaire 8 score >or=10 were considered to have current depressive symptoms. RESULTS: Our findings indicated that 95.1% of people >or=65 years old did not report current depressive symptoms and 4.9% reported current depressive symptoms. Younger adults (65-74 years), those with a high school education or less, those with an income of less than $50,000, those with good to fair self-rated health, and those with a disability were more likely to have current depressive symptoms. When stratified by age (65-74, >or=75), there was no difference in the prevalence of current depressive symptoms between the age groups. However, when the authors stratified by age group both adults 65-74 years olds and those >or=75 years old, only people with worse self-rated health and those who reported a disability were more likely to have indicated current depressive symptoms. CONCLUSION: The continued collection of data on current depressive symptoms from a population based sample of older adults can be used by policymakers and public health officials to identify local health needs and burdens that can assist in planning, directing, implementing, and monitoring the effectiveness of strategies. PMID- 19307860 TI - Visual hallucinations in dementia: a prospective community-based study with autopsy. AB - OBJECTIVE: Several studies have demonstrated that specific neuropathologic features may be associated with the presence of visual hallucinations in dementia patients, but the clinical usefulness of these studies has been limited because their subjects were selected on the basis of neuropathologic findings rather than clinical presentations. This study seeks to investigate the demographic, clinical, and neuropathologic features of community-based dementia subjects with and without visual hallucations. DESIGN: A prospective examination of the clinical and neuropathologic correlates of visual hallucinations in community based dementia subjects. PARTICIPANTS: One hundred forty-eight subjects with sufficient clinical and neuropathologic data from a community-based incident dementia autopsy case series. RESULTS: Subjects were classified according to the presence or absence of visual hallucinations and subjects with visual hallucinations (N = 27) were younger at intake and more likely to exhibit agitation, delusions, and apathy than subjects without visual hallucinations (N = 121). Subjects with visual hallucinations were also more likely than subjects without visual hallucinations to have Lewy-related pathology (LRP) (78% versus 45%). In addition, a higher frequency of visual hallucinations was observed in subjects with neocortical LRP than subjects with limbic-, amygdala-, or brainstem predominant LRP. Although Alzheimer disease with concomitant LRP was the most common neuropathologic subtype in the visual hallucinations-positive group (59%), the frequency of subjects with Alzheimer disease pathology did not differ significantly between those with and without visual hallucinations (74% versus 62%). CONCLUSIONS: Subjects with visual hallucinations were more likely to have concomitant postural and gait disturbance, additional neuropsychiatric symptoms, and neocortical LRP than subjects without visual hallucinations. Visual hallucinations accompanying dementia have distinct clinical and neuropathologic characteristics that are important for prognosis and clinical management. PMID- 19307862 TI - Personality traits influence clinical outcome in day hospital-treated elderly depressed patients. AB - OBJECTIVES: Although personality traits are considered significant predictors of both physical and mental health, their specific impact on treatment outcome in elderly patients with depression remains largely unexplored. Impact of personality traits on the evolution of depressive symptoms, quality of life, and perception of clinical progress was assessed in a psychotherapeutic community. DESIGN: A prospective longitudinal study was conducted in 62 elderly outpatients. SETTING: Day hospital treatment as usual combined group and individual therapies, pharmacological treatment, as well as family and network meetings. PARTICIPANTS: Patients presented with major depression or a depressive episode of bipolar disease. MEASUREMENTS: The Geriatric Depression Scale, the Short Form Survey, and the Therapeutic Community Assessment scale were administrated at admission, 3, 6, 12 months, and at discharge. Personality was evaluated with the NEO Five-Factor Personality Inventory. RESULTS: Outcome revealed reduced depression and improved mental quality of life and clinical progress. Higher Geriatric Depression Scale scores were found in individuals with higher levels of Neuroticism (and its Vulnerability facet). Better self-perception of clinical progress was observed in individuals with lower levels of the Depressiveness and Modesty facets and higher openness to action. Improvement in quality of life was predicted by high Positive emotions facet. All these associations remained significant after controlling for age, gender, and treatment length. CONCLUSION: Personality traits may predict clinical outcome in psychotherapeutic hospital day care for elderly patients with depression. PMID- 19307863 TI - Depression, cognition, apolipoprotein e genotype: latent class approach to identifying subtype. AB - OBJECTIVES: Possessing the epsilon4 allele of apolipoprotein E (APOE-epsilon4) genotype is associated with cognitive impairment in nondemented older adults. The authors hypothesized that they might find a subtype of depression related to impaired cognitive performance associated with the APOE-epsilon4 allele. DESIGN: A survey conducted between 2001 and 2003 with APOE genotyping. SETTING: Primary care offices in the Baltimore area. PARTICIPANTS: The study sample consisted of 305 adults aged 65 or older with complete information on APOE genotyping and covariates. MEASUREMENTS: The authors used the latent class model to classify respondents according to symptom criteria of American Psychiatric Association's Diagnostic and Statistical Manual as assessed in the Composite International Diagnostic Interview and the following four measures of cognitive function: the Mini-Mental State Exam, Hopkins Verbal Learning Test, Controlled Oral Word Association Test, and the Brief Test of Attention. The authors examined the relationship between class membership and APOE genotype. RESULTS: The latent class model yielded three classes: a nondepressed class, a class with depressive symptoms and average cognitive functioning, and a class with depressive symptoms (particularly thoughts of death and suicide) and impaired cognitive functioning. Possessing at least one APOE-epsilon4 allele was not predictive of class membership. CONCLUSION: A subgroup of elderly patients with depressive symptoms, cognitive impairment, and a high likelihood of experiencing thoughts of death or suicide may exist that may not be related to APOE-epsilon4. Subgroups of older patients with depressive symptoms may be important to identify because of the association with thoughts of death or suicide and cognitive impairment. PMID- 19307864 TI - A combination therapy of donepezil and cilostazol for patients with moderate Alzheimer disease: pilot follow-up study. PMID- 19307866 TI - Genomic discoveries in perioperative medicine: educating the audience. PMID- 19307867 TI - Gold-plated "Pender Lemon" ether device. PMID- 19307868 TI - Chloroform blues. PMID- 19307869 TI - Emerson respirator or "iron lung". PMID- 19307870 TI - Fries apparatus for artificial respiration. PMID- 19307871 TI - Why was 10% procaine used? PMID- 19307872 TI - Until proven otherwise, 5% spinal procaine is not neurotoxic. PMID- 19307873 TI - When the model does not fit the data, the model is wrong. PMID- 19307875 TI - Conventional neuromuscular monitoring versus acceleromyography: it's not the monitor but the anesthetist. PMID- 19307879 TI - Two or three interviews? PMID- 19307881 TI - Lessons from ecological and spatial studies in relation to occupational lung disease. AB - PURPOSE OF REVIEW: The majority of epidemiological research into occupational lung disease has been advanced by the study of individuals, typically in a workplace setting. This review examines how recent advances in ecological and spatial study methodology and in the information held in routine databases could be used to enhance occupational health studies. RECENT FINDINGS: Ecological studies often use routinely collected data, and this is becoming much more extensive and better validated with potential for increasing use in occupational health research. Improvements in computing power and statistical and geographical information systems methodology have led to more sophisticated mapping techniques and greater use of spatial information when investigating lung diseases usually related to occupational exposures. Ecological study methodology is experiencing a radical overhaul with supplementation of group-level data with information from small-scale individual-level studies. This hybrid design can be used to reduce bias and increase power and is directly applicable to the enhancement of aggregate information from job exposure matrices. SUMMARY: Studies of occupational lung disease can be enhanced by incorporating methodological innovations from ecological and spatial studies. PMID- 19307882 TI - Asthma, hypersensitivity pneumonitis and other respiratory diseases caused by metalworking fluids. AB - PURPOSE OF REVIEW: To highlight advances in understanding the respiratory disease associated with metal machining, a common work process involving approximately 1.2 million workers in the USA. RECENT FINDINGS: Recent studies emphasize that work-related asthma and hypersensitivity pneumonitis continue to be caused by exposure to metalworking fluid. Identification of an individual patient indicates the need for follow-up investigations at the work site to prevent additional disease and/or identify additional effected individuals. Identification of the causal agent for hypersensitivity pneumonitis has centered on microbial contamination of metalworking fluids with a number of studies focusing on Mycobacterium immunogenum. SUMMARY: Both asthma and hypersensitivity pneumonitis occur among workers exposed to metalworking fluid. The incidence of these diseases among such workers is unknown. Outbreaks of these conditions continue to be identified among metal machinists. Whether these are true outbreaks associated with some breakdown in workplace controls or, rather the recognition of ongoing endemic disease that is typically misdiagnosed as pneumonia or common adult onset asthma, needs further evaluation. Further work to elucidate the specific causal agent(s) is necessary to affect effective workplace controls. Treating an identified individual case as an index case with a follow-up workplace investigation will only be possible if practicing physicians interact with public health authorities to report newly diagnosed cases. PMID- 19307883 TI - Impact of environmental tobacco smoke and active tobacco smoking on the development and outcomes of asthma and rhinitis. AB - PURPOSE OF REVIEW: We aim to discuss current insights on the influence of active smoking and environmental tobacco smoke in lower and upper respiratory inflammatory illnesses. RECENT FINDINGS: Insight has been gained on the effect of tobacco smoking on the development of asthma from the womb to adolescence. Secondhand tobacco exposure and active smoking play a major role not only in the inception of asthma epidemiological community studies but also in patients already suffering from allergic rhinitis. Tobacco seems to influence innate immunity predisposing to Th2-associated respiratory diseases and increasing the risk for IgE-mediated sensitization. Tobacco smoking is related to worst outcomes in both asthma and rhinitis. SUMMARY: Several deleterious effects have been described in asthma because of smoking: accelerated decline in lung function, more severe symptoms, impairment in quality of life and diminished therapeutic response to steroids. The harmful effect of tobacco smoking is not only on asthma but also on rhinitis playing a role in disease outcomes. Tobacco exposure can influence innate immunity diminishing innate production of antigen-presenting cells cytokines, as well as an impaired response to toll-like receptor ligands. Active smoking is associated with current symptoms of asthma and rhinitis and seems to be a risk factor for developing new asthma in patients with rhinitis. Tobacco smoking has been also found among the factors inducing nasal obstruction and decreased muco-ciliary clearance in nonallergic rhinitis. PMID- 19307884 TI - Asthma in preschool children: the next challenge. AB - PURPOSE OF REVIEW: To describe a prospective classification for preschool wheezers according to temporal symptom pattern, and summarize findings relating to the management of viral wheeze and the use of short-term therapy for intermittent severe wheeze. RECENT FINDINGS: Phenotypes defined from cohort studies should only be applied retrospectively at school age. A new classification that can be applied prospectively is discussed. The importance of early rhinovirus-induced wheezing as a risk factor for asthma has become apparent. However, there is no benefit from short-course oral steroids for acute viral wheeze in the majority of cases. There is conflicting evidence for the role of intermittent montelukast or inhaled steroids in the treatment of acute, intermittent wheeze. A link between reduced vitamin D intake during pregnancy and increased preschool wheeze in offspring has emerged, suggesting a potential role for vitamin D supplementation in primary prevention. SUMMARY: On the basis of current evidence, a trial of bronchodilators is first-line therapy for viral wheeze, and maintenance montelukast or inhaled steroids may be considered in preschool wheezers with persistent symptoms and risk factors for future asthma. No disease-modifying therapies are available. New therapeutic options for preschool wheezing disorders are desperately needed. PMID- 19307885 TI - Natural history of lung function in health and diseases. AB - PURPOSE OF REVIEW: To outline major advances in the understanding of factors that influence lung function development through childhood. RECENT FINDINGS: New study approaches such as adjusting for 'tracking' or analysing without predefined phenotypes suggest that reduced lung function reported with several pre or coexisting features such as lower respiratory tract infections and early allergic sensitization may be spurious rather than causative. Also, two large, recent studies have clearly demonstrated that living close to major roads causes significant lung function deficits in school children, with the possible long term impact this can have on health in adult life. Furthermore, it is becoming clear that we need to focus upon early life events that can cause harm as well as have a potential for catch-up growth or development in postnatal life. SUMMARY: The implications of these findings are clearly that there is a potential for intervening in a potential pathological development. Furthermore, there is a clear need to focus research upon early life events that can improve lung growth in the damaged lung and prevent damage to the potentially healthy lung at the very start of life. PMID- 19307886 TI - Rhinosinusitis in children and asthma severity. AB - PURPOSE OF REVIEW: Rhinosinusitis is a common condition in children. The association between rhinosinusitis and asthma is supported by strong epidemiological and pathogenic evidence. Moreover, a close relationship between sinusitis and asthma severity has also been reported. This study shows the new findings in this strong relationship. RECENT FINDINGS: The profile of inflammatory cells and inflammatory mediators seen in both conditions bears considerable similarity, especially in both diseases with a relevant role played by eosinophils. SUMMARY: Severe asthma is uncommon in childhood, but there is compelling evidence showing that the most severe asthma is closely associated to upper respiratory illness, especially rhinosinusitis. Treatment of chronic rhinosinusitis, medical or surgical or both, benefits concomitant asthma and has been shown to reduce the severity of asthma. PMID- 19307887 TI - Current world literature. PMID- 19307889 TI - Role of the microcirculation in intensive care and resuscitation: the devil is in the details. PMID- 19307890 TI - The impact of the glycocalyx on microcirculatory oxygen distribution in critical illness. AB - PURPOSE OF REVIEW: Main problems of critical illness and sepsis are an altered oxygen distribution and microvascular dysfunction linked to tissue oedema. This review seeks to analyse the role of the endothelial glycocalyx in this context. RECENT FINDINGS: The presence of vascular leakage is typically associated with interstitial oedema, arterial hypotension, hypovolaemia and often a bad outcome in patients with systemic inflammation. Early goal-directed therapy provides significant benefits in severe sepsis and septic shock, but is mostly aimed at improving macrohaemodynamics. Recent data suggest that microcirculation also contributes significantly to the pathophysiology of critical illness. In fact, the endothelial glycocalyx plays a major role in vascular barrier competence. According to experimental evidence, it can easily be degraded in the presence of inflammation, but, theoretically also protected by several measures. Clinical studies revealed a positive correlation of the severity of sepsis and ischaemia with mortality, but also with a deterioration of the endothelial glycocalyx. Future investigation should focus on the preservation of this structure and assess microcirculatory variables to judge the success of cardiocirculatory therapy. SUMMARY: Deterioration of the endothelial glycocalyx initiates a breakdown of the vascular barrier in systemic inflammatory response syndrome and sepsis. Preserving this structure in critical illness might be a future therapeutical goal to improve microcirculatory oxygen distribution. PMID- 19307891 TI - Microcirculatory effects of intravenous fluids in critical illness: plasma expansion beyond crystalloids and colloids. AB - PURPOSE OF REVIEW: Plasma expanders are reviewed to determine their ability to restore microvascular function as a means for extending the transfusion trigger and delaying the use of blood transfusions. This outcome is currently achievable because of the emergence of a new understanding of optimal tissue function that prioritizes maintenance of functional capillary density, which results from the normalization of blood viscosity via the increase in plasma viscosity with new viscogenic colloids. RECENT FINDINGS: Use of viscous plasma expanders in experimental models of extreme hemodilution, hemorrhagic shock and endotoxemia shows that the limiting factor in anemia is not oxygen-carrying capacity but the decline of microvascular function due to the lowering of functional capillary density. In support of this hypothesis, we find that viscogenic colloids including high-molecular-weight starches, dextrans, polyvinylpyrrolidone, keratin and polyethylene glycol-conjugated albumin maintain or restore microvascular function in extreme hemodilution, polyethylene glycol-conjugated albumin yielding the best results. SUMMARY: Preclinical studies show that polyethylene glycol conjugated albumin at concentrations in the range of 2-4% extends the transfusion trigger, providing the more extended and complete microvascular and systemic recovery from hemorrhagic shock, extreme hemodilution and endotoxemia, postponing the need of reestablish intrinsic blood oxygen-carrying capacity to hemoglobin concentrations lower than those associated with accepted transfusion triggers. PMID- 19307892 TI - Effects of pharmaconutrients on cellular dysfunction and the microcirculation in critical illness. AB - PURPOSE OF REVIEW: A growing body of data has revealed that specific nutrient deficiencies contribute to microvascular and cellular dysfunction following critical illness. Further, targeted administration of these 'pharmaconutrients' may reverse or improve this dysfunction and improve clinical outcome. RECENT FINDINGS: Specific nutrient therapy with glutamine protects cellular metabolism and vascular function via induction of heat shock proteins, which are key proteins found to be deficient following acute illness. Arginine becomes rapidly deficient following trauma and surgery. This leads to significant immunosuppression, which when treated by arginine administration significantly reduces postoperative infection. Omega-3 fatty acids attenuate the inflammatory response and provide for resolution of ongoing inflammatory injury via production of resolvins/protectins. Antioxidants (vitamin C and selenium) and trace elements (zinc) become rapidly depleted in critical illness and replacement appears vital to ensure optimal cellular and microvascular function. Data on targeted metabolic (mitochondrial) therapies (i.e. co-enzyme Q10) show promise to improve myocardial function following cardiac surgery. SUMMARY: These specific nutrients have newly discovered vital mechanistic roles in the optimization of cellular and microcirculatory function in critical illness and injury. A growing body of literature is demonstrating that correction of key nutrient deficiencies via therapeutic administration of these pharmaconutrients can improve clinical outcome in critically ill patients. PMID- 19307893 TI - Microcirculation and mitochondria in sepsis: getting out of breath. AB - PURPOSE OF REVIEW: To present the recent findings obtained in clinical and experimental studies examining microcirculatory alterations in sepsis, their link to mitochondrial dysfunction, and current knowledge regarding the impact of these alterations on the outcome of septic patients. RECENT FINDINGS: Interlinked by a mutual cascade effect and driven by the host-pathogen interaction, microcirculatory and mitochondrial functions are impaired during sepsis. Mitochondrial respiration seems to evolve during the course of sepsis, demonstrating a change from reversible to irreversible inhibition. The spatiotemporal heterogeneity of microcirculatory and mitochondrial dysfunction suggests that these processes may be compartmentalized. Although a causal relationship between mitochondrial and microcirculatory dysfunction and organ failure in sepsis is supported by an increasing number of studies, adaptive processes have also emerged as part of microcirculatory and mitochondrial alterations. Treatments for improving or preserving microcirculatory, mitochondrial function, or both seem to yield a better outcome in patients. SUMMARY: Even though there is evidence that microcirculatory and mitochondrial dysfunction plays a role in the development of sepsis-induced organ failure, their interaction and respective contribution to the disease remains poorly understood. Future research is necessary to better define such relationships in order to identify therapeutic targets and refine treatment strategies. PMID- 19307894 TI - Should I get a Master of Business Administration? The anesthesiologist with education training: training options and professional opportunities. AB - PURPOSE OF REVIEW: Many physicians want to know whether they should get a Master of Business Administration (MBA), what type of program is best, and what career paths exist. RECENT FINDINGS: It is commonly (incorrectly) assumed that a physician successful in clinical practice can easily transfer to managing/leading an organization. To be effective, the MD/MBA must bridge the cultures of the business world and medicine. Often just a single management course is sufficient to give the physician the knowledge they seek. MBA programs come in many forms and require choosing from a range of time commitments. Leaving a good clinical job in favor of the less-defined course of an MD/MBA can be daunting. Although a wide spectrum of opportunities are available, the MD/MBA may have to start over professionally, most likely with a pay cut, and will have to 'work their way up' again. A stigma exists for MD/MBAs because they are often perceived as caring more about business than about patients. Many MD/MBAs eventually choose to stay in full-time medical practice because financial and geographic stability may be more easily attained. SUMMARY: The MBA is a good idea for the physicians who enjoy the intellectual challenges of business administration and proactively plan their own career. PMID- 19307895 TI - Current world literature. Intensive care and resucitation. PMID- 19307897 TI - Current world literature. PMID- 19307899 TI - Diagnosis and management of nocturnal enuresis. AB - PURPOSE OF REVIEW: To highlight recent advances in enuresis research. RECENT FINDINGS: Many discoveries have distanced us from the time when bedwetting was considered a psychiatric disorder. Instead, it is now agreed that nocturnal polyuria, detrusor overactivity, and high arousal thresholds are, in various combinations, central to enuresis pathogenesis. All three mechanisms have been further elucidated during the last year. It has been found that solute diuresis, and not just free-water diuresis due to vasopressin deficiency, may be causative. Sonographical bladder wall measurements have been shown to have prognostic value in detrusor-dependent enuresis, and fascinating proof for the interplay between the bladder and the sleeping brain has been put forward. And, ironically, sleep research has caused psychiatry to make a slight comeback, as studies have indicated that enuretic children may suffer from cognitive problems due to suboptimal sleep. Less has been achieved regarding treatment, but some evidence has finally supported the experience that anticholinergics may be effective in therapy-resistant enuresis. SUMMARY: During the last years, increased insight has been gained into the multifaceted pathogenesis of enuresis, but there is still an irritating lack of proven effective therapies. PMID- 19307900 TI - Diagnosis and management of hypercalciuria in children. AB - PURPOSE OF REVIEW: In this review, recent advances in the epidemiology, genetics, clinical associations and management of idiopathic hypercalciuria will be discussed. RECENT FINDINGS: A significant genetic contribution exists in the pathophysiology of hypercalciuria. Although several candidate genes and genetic alterations have been proposed, identification of precise gene(s) responsible remains elusive. Decreased bone density has been increasingly associated with hypercalciuria. Recent publications have suggested that bisphosphonates may play a role in the management in patients in whom both hypercalciuria and decreased bone density are present. SUMMARY: Idiopathic hypercalciuria is a common disorder in children and can present with a range of clinical presentations such as hematuria, voiding dysfunction, flank pain, abdominal pain, nephrolithiasis, urinary tract infection and decreased bone mineral density. Dietary modifications are often sufficient in the management of hypercalciuria. If the symptoms persist or a rare monogenic disorder is present, consideration should be given to medical treatment with a thiazide diuretic and/or citrate therapy. PMID- 19307901 TI - Current world literature. PMID- 19307925 TI - Does quality of observational studies affect the results of a meta-analysis?: the case of cigarette smoking and pancreatic cancer. AB - OBJECTIVES: A meta-analysis of observational studies on association between cigarette smoking and pancreatic cancer was performed to focus, particularly, on the role of the studies' quality in affecting meta-analysis results. METHODS: A bibliographic search was carried out on PubMed and EMBASE databases until February 15, 2008. Key words were "pancreatic neoplasms," "pancreatic cancer," "smoking," "smoke," "cigarette," "case-control studies," and "cohort studies." Studies about cigarette smoking and pancreatic cancer were selected and assessed on quality. STATA 9.0 was used. RESULTS: Six cohort studies and 24 case-control studies were selected, with median quality scores of 8 (range, 3) and 10 (range, 8), respectively. Pooled case-control studies' odds ratio (OR) and cohort studies' risk ratio were, respectively, 1.45 (95% confidence interval [CI], 1.33 1.57) and 1.78 (95% CI, 1.64-1.92). After stratifying for quality scoring, high quality-scored case-control studies yielded an OR of 1.38 (95% CI, 1.27-1.49), whereas the others gave an OR of 1.52 (95% CI, 1.34-1.73). The results of meta analysis for cohort studies showed a risk ratio of 1.74 (95% CI, 1.61-1.90) and of 2.10 (95% CI, 1.64-2.67), respectively, for high- and low-quality score studies. CONCLUSIONS: There is evidence that cigarette smoking is an important risk factor for pancreatic cancer, but the estimate of the association greatly relies on the studies' quality. PMID- 19307926 TI - Neurogenin 3 and neurogenic differentiation 1 are retained in the cytoplasm of multiple endocrine neoplasia type 1 islet and pancreatic endocrine tumor cells. AB - OBJECTIVES: To investigate if transcription factors involved in pancreatic differentiation and regeneration are present in pancreatic endocrine tumors and if they are differentially expressed in normal pancreas compared with multiple endocrine neoplasia type 1 (MEN1) nontumorous pancreas. METHODS: The expression of neurogenin 3 (NEUROG3), neurogenic differentiation 1 (NEUROD1), POU class 3 homeobox 4 (POU3F4), pancreatic duodenal homeobox factor 1 (PDX1), ribosomal protein L10 (RPL10), delta-like 1 homolog (Drosophila; DLK1), and menin was analyzed by immunohistochemistry in normal pancreas and pancreatic endocrine tumors from 6 patients with MEN1 and 16 patients with sporadic tumors, as well as pancreatic specimens from Men1 heterozygous and wild type mice. Quantitative polymerase chain reaction was performed in a subset of human tumors. RESULTS: Tumors and MEN1 nontumorous endocrine cells showed a prominent cytoplasmatic NEUROG3 and NEUROD1 expression. These factors were significantly more expressed in the cytoplasm of Men1 heterozygous mouse islet cells compared with wild type islets; the latter showed an exclusively nuclear reactivity. The degree of Pou3f4, Rpl10, and Dlk1 immunoreactivities differed significantly between islets of heterozygous and wild type mice. The expressions of RPL10 and NEUROD1 were prominent in the MEN1 human and heterozygous mouse exocrine pancreas. Insulinomas had significantly higher PDX1 and DLK1 messenger RNA levels compared with other tumor types. CONCLUSIONS: Transcription factors involved in pancreatic development show altered expression and subcellular localization in MEN1 nontumorous pancreas and pancreatic endocrine tumors. PMID- 19307928 TI - Loss of expression of E-cadherin in solid pseudopapillary tumors of the pancreas. PMID- 19307929 TI - D2-40 immunocytochemical staining for pancreatic islets and pancreatic endocrine tumors. PMID- 19307930 TI - Pancreatic endocrine tumor in multiple endocrine neoplasia type 1 syndrome with intraductal growth into the main pancreatic duct. PMID- 19307931 TI - Islet amyloidosis and beta-cell neogenesis in chronic calcifying pancreatitis with non-insulin-dependent diabetes mellitus. PMID- 19307932 TI - Pancreatitis induced in rats by repetitive administration of L-arginine. PMID- 19307933 TI - Sclerosing cholangitis, autoimmune chronic pancreatitis, and situs viscerum inversus totalis. PMID- 19307934 TI - Slow growth of small pancreatic carcinoma with a 20-04 follow-up. PMID- 19307935 TI - A case of squamoid cyst of pancreatic ducts. PMID- 19307936 TI - Acute recurrent pancreatitis by heterotopic fundic mucosa at the ampulla of vater. PMID- 19307937 TI - Pharmacokinetics of mycophenolic acid and its glucuronide metabolites in stable adult liver transplant recipients with renal dysfunction on a low-dose calcineurin inhibitor regimen and mycophenolate mofetil. AB - Low-dose calcineurin inhibitors (CNIs) in combination with a fixed dose (2 g/d) of mycophenolate mofetil (MMF) are a strategy to minimize exposure to cyclosporine (CSA) or tacrolimus (TAC) and thus reduce CNI-related side effects. This study compared the pharmacokinetics (PK) of mycophenolic acid (MPA) and its glucuronide metabolites in stable adult liver transplant recipients with moderately impaired renal function converted from a standard to a low-dose CNI regimen in combination with a fixed dose of MMF. Full 12-hour PK profiles of MPA, free MPA, the aryl glucuronide (MPAG), and the acyl glucuronide (AcMPAG) were obtained from 30 stable liver transplant patients on low-dose CNI (CSA, n = 12; TAC, n = 18) therapy at least 3 months after initiation of low-dose therapy. Predose CSA and TAC concentrations (quantified by liquid chromatography-tandem mass spectrometry) ranged from 17 to 35 and 1.1 to 3.7 microg/L, respectively. The PK variables for MPA, MPAG, AcMPAG, and free MPA displayed wide interindividual variability. Of note was the observation that there were no significant differences in the exposure to MPA, MPAG, and free MPA between the CSA and TAC groups. MPA area under the concentration-time curves (AUCs) ranged from 31.8 to 102.1 (median: 52.9) mg.h(-1).L(-1) in the CSA group and from 22.9 to 144.8 (median: 55.9) mg.h(-1).L(-1) in the TAC group. The AcMPAG AUC on patients under low-dose CSA therapy was higher than that observed under patients on low-dose TAC therapy, although this did not quite reach statistical significance (P = 0.057). Patients receiving CSA had a significantly higher AcMPAG Cmax but not AcMPAG AUC, suggesting that only peak CSA concentrations on a low-dose CSA regimen are sufficient to impair the biliary excretion of AcMPAG. In summary, the influence of CSA on the exposure to MPA was attenuated in stable adult liver transplant recipients on a low-dose CNI therapy in combination with a fixed dose of MMF as compared with patients on a standard CNI therapy. Dose adjustment according to drug concentration measurements is recommended to optimize dosing of MMF and to maintain adequate immunosuppression in patients converted to low-dose CNI therapy. PMID- 19307938 TI - A high-throughput assay using liquid chromatography-tandem mass spectrometry for simultaneous in vivo phenotyping of 5 major cytochrome p450 enzymes in patients. AB - The phenotyping cocktail is a practical approach for phenotyping of cytochrome P450 (CYP) enzymes in vivo. In this study, a liquid chromatography-tandem mass spectrometry method using a dual-extraction approach was developed and validated to quantify 5 selective substrates and their metabolites for the simultaneous phenotyping CYPs 1A2, 2C19, 2C9, 2D6, and 3A4 in patient blood samples. The assay was applied in a pilot study of 11 patients with schizophrenia. Five blood samples were collected before and at 1, 2, 4, and 6 hours after administration of a phenotyping cocktail consisting of 100 mg caffeine, 20 mg omeprazole, 25 mg losartan, 30 mg dextromethorphan, and 2 mg midazolam. The method successfully quantitated the CYP enzyme activities without serious side effects in patients. The ratios of metabolite to parent area under the concentration-time curve values were calculated over the 6-hour postdosage to reflect CYP2D6, CYP3A4, and CYP2C9 activities. The ratios of metabolite to parent plasma concentrations were calculated at 4-hour postdosage for CYP1A2 and at 4- or 6-hour postdose for CYP2C19, respectively. The plasma concentration of midazolam at 4 hours was also estimated as another phenotyping index for CYP3A4 activity. The simultaneous assay of all these analytes in a single matrix (plasma) will increase the feasibility of CYP phenotyping in patients. PMID- 19307939 TI - Stability of drugs of abuse in oral fluid collection devices with purpose of external quality assessment schemes. AB - As the stability of drugs of abuse in oral fluid can affect drug testing results, we evaluated this parameter together with recovery for the principal drugs of abuse in two collection devices typically used to ship oral fluid samples to testing laboratories. Two different samples were prepared using Cozart Drug Detection System and Intercept oral fluid collection devices with 600 ng/mL of 6 monoacetylmorphine (6-MAM) and cocaine and 240 ng/mL of Delta tetrahydrocannabinol (THC) and 11-nor-9-carboxy-Delta-tetrahydrocannabinol (THCCOOH). Samples were sent at ambient temperature by courier to the participating laboratories (n = 19) the same day of preparation. Samples were analyzed upon reception (about 48-72 hours after shipment). Percent coefficients of variation, calculated using robust mean and robust standard deviation, were around 30% for all analytes, except for THCCOOH in both samples (between 50% and 80%) and THC in 1 sample (50%). Percent coefficients of variation were also high (between 50% and 70%) for morphine and benzoylecgonine, formed after 6-MAM and cocaine spontaneous hydrolysis. On average, 9%-12% 6-MAM was converted to morphine and between 26% and 41% cocaine to benzoylecgonine. Good recoveries were observed for the acid metabolite of THC in both collection devices, whereas THC was always scarcely recovered. Depending on the collection device used, obtained results may confound the interpretation and estimation of blood drug concentration, given an oral fluid drug concentration and subsequent consideration of time elapsing from drug consumption. PMID- 19307940 TI - The utility of nucleos(t)ide-only regimens in the treatment of Mycobacterium tuberculosis-HIV-1 coinfection. AB - HIV-tuberculosis coinfection is complex partly because rifamycins reduce therapeutic levels of protease inhibitors and nonnucleoside reverse transcriptase inhibitors, leading to potential virological failure. One therapeutic option is to use nucleos(t)ide-only regimens that have minimal interactions with antituberculous therapy. We report the largest published series of HIV tuberculosis coinfected patients successfully treated with nucleos(t)ide regimens and antituberculous therapy. This group achieved similar virological and immunological outcomes when compared with tuberculosis patients on nonnucleoside reverse transcriptase inhibitor or protease inhibitor-based HAART, demonstrating the utility of this approach. PMID- 19307942 TI - Broad usage spectrum of G protein-coupled receptors as coreceptors by primary isolates of HIV. AB - OBJECTIVE: HIV-1 can use various G protein-coupled receptors (GPCRs) in addition to CCR5 and CXCR4 as coreceptors; however, this type of HIV-1 infection has hardly been detected in vivo. The objective of this study was to elucidate the spectrum of GPCR usage by HIV-1 populations in vivo. DESIGN: CD4-expressing glioma cell line, NP-2/CD4, becomes highly susceptible to HIV-1 when the cells express GPCRs with coreceptor activities. This cell system was advantageous for detecting the inefficient use of GPCRs by HIV-1. METHODS: We developed NP 2/CD4/GPCR cells that express each of 23 GPCRs: 21 chemokine receptors (CCR1, CCR2b, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9B, CCR10, CCR11, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CX3CR1, XCR1, D6, and DARC) and two other GPCRs (a formylpeptide receptor, FPRL1, and an orphan GPCR, GPR1). NP-2/CD4/GPCR cells were directly cocultured with HIV-1-positive peripheral blood lymphocytes and HIV 1 infection was detected. RESULTS: Primary HIV-1 isolates were obtained from NP 2/CD4/GPCR cells expressing CCR5, CXCR4, FPRL1, or GPR1 cocultured with 11 of 17 peripheral blood lymphocytes. Surprisingly, these isolates showed extremely expanded GPCR usage, such as CCR1, CCR3, CCR5, CCR8, CXCR4, D6, FPRL1, and GPR1 as coreceptors. We found that CCR9B, CCR10, and XCR1 also work as novel HIV-1 coreceptors. CONCLUSION: FPRL1 and GPR1 have the potential to work as significant HIV-1 coreceptors in vivo next to CCR5 and CXCR4. HIV-1 populations that can use various GPCRs as coreceptors are already circulating in vivo, even in the early stage of HIV-1 infection. PMID- 19307943 TI - Is it smart to continue to study treatment interruptions? PMID- 19307941 TI - Tolerance and viral resistance after single-dose nevirapine with tenofovir and emtricitabine to prevent vertical transmission of HIV-1. AB - OBJECTIVE: Viral resistance occurs with high frequency after single-dose nevirapine. We aimed to evaluate the safety and resistance profiles of a combination of tenofovir disoproxil fumarate (TDF) and emtricitabine (FTC) in HIV 1-infected pregnant women and their newborns. DESIGN: An open-label phase I/II trial in Cote d'Ivoire, Cambodia and South Africa. METHODS: Women received antenatal zidovudine, intrapartum single-dose nevirapine and two tablets of TDF/FTC and one daily tablet of TDF/FTC during the 7 days postpartum. Their infants received single-dose nevirapine and zidovudine for 1 week. Serious adverse events (SAEs), kinetic of maternal plasma HIV-1 RNA viral load, genotypic resistance at 28 days postpartum and paediatric HIV-1 infection at 3, 28 and 45 days of life were assessed. RESULTS: Thirty-eight HIV-1-infected pregnant women were enrolled (19 in Abidjan, 12 in Phnom Penh and seven in Soweto) with a median CD4 cell count of 450 cells/microl and median viral load of 4.08 log10 copies/ml. Women received TDF/FTC 4.9 h in median before delivery. Biological SAEs occurred in nine women. Among 39 live births, nine infants had clinical SAEs, including four deaths, and two developed severe anaemia. These SAEs were not likely to be related to TDF/FTC. Maternal viral load decreased by a median of 0.90 log10 copies/ml at 2 days postpartum and returned to baseline value at 28 days. No intrapartum HIV transmission was reported. No genotypic resistance mutation to zidovudine, nevirapine, FTC or TDF was detected. CONCLUSION: The TDF/FTC combination was well tolerated in delivering women and exposed newborns. Nevirapine viral resistance appears to have been avoided by the intrapartum and 7 day postpartum TDF/FTC regimen. PMID- 19307944 TI - Prognostic significance of interaction between somatic APC mutations and 5 fluorouracil adjuvant chemotherapy in Taiwanese colorectal cancer subjects. AB - OBJECTIVE: The correlations between adenomatous polyposis coli (APC) mutations and 5-fluorouracil (5-FU) adjuvant chemotherapy and colorectal cancer (CRC) patients' prognosis are not well known. We performed an exploratory study to investigate the association between APC mutations and the survival of Taiwanese CRC subjects who received 5-FU adjuvant chemotherapy. METHODS: Full-length APC gene isolated from tumor tissue and adjacent normal colon tissue from 117 CRC subjects was sequenced. Various characteristics of the 117 subjects were recorded and used in the Cox proportionalhazard model analyses. RESULTS: Although the subject survival rate was associated with the cancer stage, but not with the occurrence of APC mutations, we demonstrate a significant interaction between the somatic APC mutations and 5-FU adjuvant chemotherapy to the prognosis of CRC subjects. Subjects carrying APC mutation(s) and receiving 5-FU adjuvant chemotherapy demonstrate increased hazards (vs. no APC mutation or chemotherapy) for all cause (hazard ratios = 5.565; P = 0.042) or CRC deaths (hazard ratios = 6.920; P = 0.043). 5-FU adjuvant chemotherapy only decreases hazards in CRC subjects without APC mutation(s) for all cause death (hazard ratios = 0.257; P = 0.003) or CRC death (hazard ratios = 0.342; P = 0.028). CONCLUSIONS: 5-FU adjuvant chemotherapy only prevents CRC subjects without somatic APC mutation(s) from all cause death or CRC death. It needs further studies with larger sample size and longer follow-up time to confirm these results. PMID- 19307945 TI - Results of a phase I trial of 12 patients with locally advanced pancreatic carcinoma combining gefitinib, paclitaxel, and 3-dimensional conformal radiation: report of toxicity and evaluation of circulating K-ras as a potential biomarker of response to therapy. AB - OBJECTIVE: To evaluate the toxicity of daily gefitinib, an epidermal growth factor receptor-tyrosine kinase inhibitor, with concurrent chemoradiation (CRT) in patients with locally advanced pancreatic adenocarcinoma and prospectively evaluate plasma k-ras as a potential marker of response to gefitinib and CRT. METHODS: Eleven of 12 eligible patients enrolled received a 7-day induction of gefitinib (250 mg PO) followed by daily gefitinib with concurrent CRT. Patients received 50.4 Gy/28 fractions of external beam radiation with weekly paclitaxel (40 mg/m IV) followed by maintenance on gefitinib. Plasma k-ras codon 12 mutations were detected using a two-stage restriction fragment length polymorphism-polymerase chain reaction assay on patients' plasma both before and after therapy. Mutations were confirmed by direct sequencing. RESULTS: Common adverse events included grade 1 skin rash (63%), grade 1 to 2 gastrointestinal symptoms including anorexia, nausea, vomiting, and diarrhea occurred in 63% of patients, grade 3 nausea occurred in 45% of patients. Three patients did not complete therapy, only one was possibly associated with study drug. K-ras mutations were detected in the pre-gefitinib plasma of 5/11 patients and in the matched tumor tissue of 3/4 patients. In patients where k-ras mutations were undetectable post-treatment, survival times were favorable. CONCLUSIONS: Combination of daily gefitinib with concurrent CRT in this locally advanced pancreatic cancer population was reasonably tolerated. Rapid changes in serum k ras may provide critical information as to the efficacy of a novel agent and assist in tailoring treatment for cancers of the pancreas. PMID- 19307946 TI - Familial breast cancer: clinical response to induction chemotherapy or radiotherapy related to BRCA1/2 mutations status. AB - PURPOSE: BRCA1/2 germline mutations are associated with impaired DNA double strand break repair. We tested whether breast cancers in BRCA1/2 mutation carriers were more responsive to induction treatments than in noncarriers. PATIENTS AND METHODS: The BRCA1 and BRCA2 genes were screened for germline mutation in a retrospective cohort of 90 patients (with 93 tumors) with a family history of breast and/or ovarian cancer, treated with induction anthracycline containing chemotherapy and/or radiotherapy. Median tumor size was 40 mm. Clinical responses and breast preservation rates were correlated to BRCA1/2 mutation status, and to other clinical and pathologic factors. RESULTS: A complete clinical response was achieved in 15/39 (46%) BRCA1/2-mutated tumors and in 7/54 (17%) nonmutated tumors (P = 0.008). Complete or major clinical response rate was observed in 55 of the 74 tumors treated with induction chemotherapy (74.3%). The overall complete or major clinical response rate in the tumors treated with induction radiotherapy was 68% (13/19 tumors). Following induction treatment by either chemotherapy or radiotherapy, more breast-conserving treatments could be performed in mutation carriers than in noncarriers: the rates of breast preservation were 82% in BRCA1/2-mutated tumors and 63% in nonmutated tumors, respectively (P = 0.045). BRCA1 mutation was the sole predictor of breast conservation. CONCLUSION: Breast conservation after induction treatment was higher in BRmut+ tumors, and clinical response was related to aggressive tumor features correlated with BRCA1/2 mutations. This suggests that impaired repair mechanisms related to the BRCA1/2 mutations increased the chemosensitivity and radiosensitivity of large breast cancers. Further studies will need to determine the long-term outcome in these patients. PMID- 19307947 TI - Acute leukemia in patients sixty years of age and older: a twenty year single institution review. AB - OBJECTIVES: Acute leukemia, particularly acute myeloid leukemia, occurs more frequently in the elderly, a growing segment of the North American population. To evaluate our progress in the diagnosis, treatment and outcome of this condition, we reviewed our experience of all patients > or =60 years of age diagnosed with acute leukemia over a 20-year period at Saint Paul's Hospital, a university-based hospital in Vancouver, Canada. METHODS: A retrospective chart review was performed of 103 patients > or =60 years of age diagnosed with acute leukemia (acute myeloid leukemia-81; acute lymphoid leukemia-15; acute leukemia not otherwise specified-7). RESULTS: Median age was 72 (range 60-88) years. Bone marrow aspirate yielded cytogenetic information on 57 patients and 18 (31.6%) had an unfavourable karyotype. Fifty-three (51%) patients received induction chemotherapy (treated) and 50 (49%) were palliated (untreated). Treated patients were younger [median 67 years (range 60-79)] than untreated patients [76 years (61-88)], (P < 0.0001). Of the treated patients, 33 (62%) achieved a complete remission. The median overall survival for the group was 104 (1-2689) days, and for treated versus untreated patients-219 (1-2689) and 39 (2-1229) days, respectively (P = 0.0021). Univariate variables predictive of prolonged survival included induction chemotherapy (P = 0.0027), de novo leukemia (P = 0.0420), and younger age, with a relative increase in death in older subgroups (60-69, 70-79, 80+), (P = 0.0311). Induction chemotherapy was the only predictor of prolonged survival in multivariate analysis (P = 0.0027). CONCLUSIONS: The prognosis of acute leukemia in older patients remains poor, and even though induction chemotherapy seem to prolong survival in patients able to receive treatment, most ultimately die of leukemia. PMID- 19307948 TI - Thyroid dysfunction in patients treated with radiotherapy for neck. AB - PURPOSE: The aim of the present study was to retrospectively evaluate the early and late changes in thyroid dysfunction after radiation therapy for head and neck cancer either with or without surgery. PATIENTS AND METHODS: Sixty-three patients receiving neck irradiation including the thyroid gland were recruited in the study. Thirty-six patients had undergone either a functional or radical neck dissection, and radiotherapy was the primary treatment in 27 patients. RESULTS: Of 63 patients, 24 (38%) were diagnosed with hypothyroidism (HT), 8 (12.7%) with clinical HT, and 16 (25.4%) with subclinical HT. The median time to the development of clinical HT was 15 months (range, 0-36 months) and subclinical HT was 3 months (range, 0-24 months). Eleven (17.5%) of the patients were diagnosed with subclinical hyperthyroidism. The median time to the development of the subclinical hyperthyroidism was 0 months (completion of radiation therapy) (range, 0-3 months). Univariate analyses of age, smoking history, neck RT dose, clinical stage, concurrent chemotherapy, and surgery failed to identify a clinically relevant risk factor for HT. Univariate analysis of clinical HT revealed that the elevated pre-radiation therapy thyroid-stimulating hormone level was significant factor (P = 0.021). CONCLUSION: HT associated with head and neck irradiation. We recommend that thyroid function should be evaluated periodically in patients who have undergone neck radiation. PMID- 19307949 TI - High dose rate and external beam radiotherapy in locally advanced prostate cancer. AB - PURPOSE: To report the outcomes and toxicity of high dose rate brachytherapy as a boost for localized prostate cancer. MATERIALS AND METHODS: Between 1997 and 2000, the medical records of 131 patients with prostate adenocarcinoma who were treated with external beam radiation therapy and high dose rate brachytherapy, were retrospectively analyzed. Furthermore, 55% of the patients received neoadjuvant/concurrent or adjuvant androgen deprivation therapy. Patients were stratified into 2 groups. Group 1 included 65 patients with Gleason score 7, pretreatment prostate specific antigen (PSA) between 10 and 20 ng/mL, and clinical stage T2b. Group 2 included 66 patients with Gleason score between 8 and 10, PSA greater than 20 ng/mL, and clinical stage greater than T2b. RESULTS: At a median follow-up of 62.8 months, the 5-year biochemical control (BC) rate, as defined by the American Society for Therapeutic Radiology and Oncology Phoenix Consensus panel statement, was 81% and overall survival was 91%. BC in Groups 1 and 2 were 87% and 71%, respectively. On univariate analysis risk group, pretreatment PSA and age were significant predictors of BC. However, on multivariate analysis only pretreatment PSA was significant. Using the Radiation therapist oncology group criteria, there were 2 (1.5%) cases of grade 3 acute urinary toxicity. Regarding late side effects (n = 5), 4% of patients had grade 3 genitourinary toxicity and no grade 4 complication was observed. CONCLUSIONS: External beam radiation therapy and high dose rate brachytherapy for prostate cancer resulted in excellent BC, and overall survival with minimal severe, acute, or late complications. PMID- 19307950 TI - Patient perspectives regarding the value of total skin electron beam therapy for cutaneous T-cell lymphoma/mycosis fungoides: a pilot study. AB - PURPOSE: Cutaneous T-cell lymphoma (CTCL) represents a group of lymphoid malignancies involving the skin with mycosis fungoides being most common. This prospective study was initiated to evaluate perceptions of total skin electron beam therapy (TSEBT) by patients with mycosis fungoides who had failed at least 3 prior therapies. MATERIALS AND METHODS: Before the start of TSEBT, at 1 month and 6 months after the completion of treatment, patients completed an anonymous questionnaire for each of the prior therapies and for TSEBT. It identified the following: (1) success of the therapy, (2) difficulty in the therapy, (3) ability to recover from the therapy, (4) risk/benefit ratio of the therapy, and (5) willingness to repeat the therapy or therapies in question. Mean values for each question were compared using the paired t test. RESULTS: Thirteen patients were enrolled in the study. At 1- and 6-months posttreatment, patients thought that TSEBT was more difficult to undergo and recover from in comparison to other therapies. At 6-months posttreatment, patients also opposed the repeat therapy with TSEBT compared with other treatments. Patients perceived TSEBT as more successful than other therapies, but this difference in perception was not statistically significant. CONCLUSIONS: TSEBT is perceived by patients to be a successful treatment. TSEBT is thought to be a difficult treatment to undergo and recover from compared with other treatments. PMID- 19307951 TI - Xerostomia in long-term survivors of aggressive non-Hodgkin's lymphoma of Waldeyer's ring: a potential role for parotid-sparing techniques? AB - BACKGROUND: The degree of xerostomia in patients treated for intermediate-and high-grade non-Hodgkin lymphoma (NHL) of Waldeyer's ring (WR) is unknown. METHODS AND MATERIALS: Fifteen patients treated for stage I-IV NHL of WR with radiotherapy (RT) were administered a xerostomia questionnaire. Numerical responses (0 = no xerostomia; 100 = maximum xerostomia) were compared with responses from 5 sets of patients treated for head and neck squamous cell carcinoma who were grouped by amount of parotid in RT field: larynx-only, ipsilateral parotid, bilateral-partial parotid, bilateral-total parotid, parotid sparing intensity-modulated radiotherapy. RESULTS: Waldeyer's patients' median xerostomia questionnaire score was 31, which was significantly different from the larynx-only group, bilateral-partial parotid group, and bilateral-total parotid group, but not significantly different from the ipsilateral parotid group or parotid-sparing intensity-modulated radiotherapy group. CONCLUSIONS: Xerostomia in survivors WR NHL is a detectable toxicity with severity like that in head and neck squamous cell carcinoma patients who receive ipsilateral parotid irradiation, and warrants parotid-sparing RT techniques. PMID- 19307952 TI - Malignant ovarian germ cell tumors: a single-institution experience. AB - OBJECTIVE: To evaluate the clinicopathologic prognostic factors in malignant ovarian germ cell tumors. METHODS: We reviewed the medical records of 70 patients treated from 1990 to 2006 at our center. Clinical data including demographics, stage, surgery, chemotherapy, survival, menses status, and fertility were collected from patients' charts. RESULTS: Median age was 22 years (range, 9-68). The histologic subtypes included 36 dysgerminomas, 11 yolk sac tumors, 3 immature teratomas, 1 embryonal carcinomas, and 19 mixed types. The most striking clinicopathologic finding was a history of concomitant immunosuppressant therapy, which was observed in 2 patients. Two patients had contralateral sex-cord tumors at presentation and follow-up. During a median follow-up period of 4.6 years, 11 patients had recurrence. The median time to recurrence was 8 months (6-28 months). Recurrences appeared in the abdominopelvic cavity in 9 out of 11 patients. Only one could be salvaged with second-line chemotherapy. Cumulative survival rate was 97% and 60% in patients with dysgerminoma and nondysgerminoma, respectively. Nondysgerminoma histology and residual tumor after surgery were unfavorable prognostic factors (P < 0.001 and P = 0.015). Fertility-sparing surgery was as effective as radical surgery among all eligible patients. Of patients with known menstrual status, 96% had regular menses. Of the 8 patients who opted for conception among these patients, 7 delivered healthy infants. CONCLUSIONS: Nondysgerminomas have an aggressive clinical course. New treatment strategies are needed for eradication of abdominopelvic disease at initial diagnosis and recurrent setting. Occurrence of malignant ovarian germ cell tumors may be associated with immunosuppression in some patients. Sex-cord stromal tumors may present with bilateral involvement. It is possible to maintain fertility after fertility-sparing surgery followed by chemotherapy. PMID- 19307953 TI - Proton therapy for prostate cancer treatment employing online image guidance and an action level threshold. AB - PURPOSE: The ability to determine the accuracy of the final prostate position within a determined action level threshold for image-guided proton therapy is unclear. MATERIALS AND METHODS: Three thousand one hundred ten images for 20 consecutive patients treated in 1 of our 3 proton prostate protocols from February to May of 2007 were analyzed. Daily kV images and patient repositioning were performed employing an action-level threshold (ALT) of > or = 2.5 mm for each beam. Isocentric orthogonal x-rays were obtained, and prostate position was defined via 3 gold markers for each patient in the 3 axes. RESULTS: To achieve and confirm our action level threshold, an average of 2 x-rays sets (median 2; range, 0-4) was taken daily for each patient. Based on our ALT, we made no corrections in 8.7% (range, 0%-54%), 1 correction in 82% (41%-98%), and 2 to 3 corrections in 9% (0-27%). No patient needed 4 or more corrections. All patients were treated with a confirmed error of < 2.5 mm for every beam delivered. After all corrections, the mean and standard deviations were: anterior-posterior (z): 0.003 +/- 0.094 cm; superior-inferior (y): 0.028 +/- 0.073 cm; and right-left (x) -0.013 +/- 0.08 cm. CONCLUSION: It is feasible to limit all final prostate positions to less than 2.5 mm employing an action level image-guided radiation therapy (IGRT) process. The residual errors after corrections were very small. PMID- 19307954 TI - Pretreatment prognostic factors of survival in patients with locally advanced nonmetastatic squamous cell carcinoma of the head and neck treated with radiation therapy with or without concurrent chemotherapy. AB - BACKGROUND: Identification of pretreatment prognostic factors influencing overall survival (OS) in locally advanced squamous cell carcinoma of the head and neck is an important issue in head and neck oncology. METHODS: A total of 289 patients were treated with standard fraction or hyperfractionated radiation therapy with or without concurrent low-dose daily chemotherapy. RESULTS: Gender (P = 0.43) and age (P = 0.26) did not influence OS whereas Karnofsky Performance Status (KPS) (P < 0.0001), T stage (P < 0.0001), and N stage (P < 0.0001) did. Stage grouping was another factor that influenced OS (P < 0.001). Patients with larynx and nasopharynx fared better than those with other primaries (P = 0.0153). Finally, treatment significantly influenced OS. Multivariate analysis showed that KPS, T and N stage, and treatment were independent prognosticators of OS. CONCLUSIONS: KPS, T and N stage, and treatment are independent prognosticators of OS in patients with locally advanced squamous cell carcinoma of the head and neck treated with radiation therapy with or without concurrent low-dose daily chemotherapy. PMID- 19307955 TI - Partial breast brachytherapy as the primary treatment for breast cancer diagnosed after mantle radiation therapy for Hodgkin's disease. AB - PURPOSE: Mastectomy is the most widely accepted treatment for managing breast cancer in patients who have previously received mantle radiation therapy (RT). With accumulating evidence supporting partial breast irradiation, we treated newly diagnosed breast cancer among lymphoma survivors who had history of RT with lumpectomy and breast brachytherapy. METHODS: Five patients, with history of Hodgkin disease (HD) underwent lumpectomy and sentinel lymphadenectomy +/- axillary dissection followed by low dose rate Ir-192 interstitial brachytherapy. A multicatheter technique was used and all patients received 45 Gy to the target volume over 4 to 5 day time interval. Adjuvant systemic therapy when indicated was administered after completion of brachytherapy. RESULTS: The median time from the treatment of the HD to the diagnosis of breast cancer was 242 months (range: 68-322 months). The median T-size of the invasive breast cancer is 0.7 cm. The range of follow up after brachytherapy is 5 to 67 months. All 5 patients have an intact breast and are without evidence of relapse. No patients developed an infection. No skin toxicity > or =grade 2 was observed. All patients have an excellent cosmetic result. No excessive fibrosis or necrosis has been observed. CONCLUSION: The preliminary experience using partial breast brachytherapy (PBB) in patients with previous history of mantle RT suggests low complication rates, and acceptable cosmetic results. The excellent local control and freedom from mastectomy for local recurrence warrants further investigation to establish brachytherapy as an acceptable alternative to mastectomy for early stage breast cancer among HD survivors with history of prior RT. PMID- 19307956 TI - The influence of intraarterial high-dose cisplatin with concomitant irradiation on arterial microanastomosis: an experimental study. AB - OBJECTIVE: In this experimental study with rabbits, the influence of intraarterial high-dose cisplatin with concomitant irradiation on arterial microanastomoses was evaluated to determine their impact on free-tissue transfers. METHODS: The right and left iliac arteries of 10 rabbits were injected with 150 mg/m of cisplatin (group 1). To serve as physiological controls, the iliac arteries of 10 other rabbits were injected with the same volume of saline (group 2). Hypofractionated radiotherapy was given to the right inguinal area of all rabbits using a Co unit, 1.25 MeV, and an SSD of 80 cm for 25 Gy at 5 fractions a day for 5 days (groups 1A and 2A) and the left inguinal areas remained unirradiated (groups 1B and 2B). Both femoral arteries of all 20 rabbits were transected and anastomosed using microsurgical techniques on day 7 after the treatment. All femoral artery anastomoses were examined under anesthesia for pulsatile blood flow 14 days after the surgery. Arteries, including the anastomotic site, were harvested and fixed for histologic evaluation by light microscopy and transmission electron microscopy. RESULTS: Microscopic evaluation showed that all femoral artery anastomoses had good, pulsatile blood flow. Histologic examination of the femoral artery anastomotic site revealed changes of the arterial walls that varied between the groups. Evidence of intimal changes included detachment of endothelial cells in the intimal layer, edema of the endothelial cells in the intima, intimal thickening, separation of the intima from the tunica media, and collagen deposition. Evidence of damage to the tunica media included vacuolation and disarray of the smooth muscle cells, fibrinoid necrosis, and hemorrhage. The damage was most pronounced in the arteries that received both intraarterial cisplatin and radiotherapy (group 1A). The degree of damage diminished in the arteries of the radiotherapy-alone group (group 2A) and the intraarterial cisplatin-alone arteries (group 1B) compared with the control arteries (group 2B). Despite the arterial damage after irradiation and/or cisplatin, the patency rates after vascular anastomosis were 100% for every group. CONCLUSIONS: Although damage to the arterial walls in the group that received intraarterial high-dose cisplatin with concomitant irradiation was most obvious, there were no differences in the patency rates after vascular anastomosis between any of the groups. Thus, after intraarterial high-dose cisplatin with concomitant irradiation, the femoral arteries can be used with caution as recipient vessels for free-tissue transfer. PMID- 19307957 TI - G3139 (Genasense) in patients with advanced merkel cell carcinoma. AB - OBJECTIVES: Merkel cell carcinoma (MCC) is a rare, aggressive neuroendocrine malignancy of the skin. Preclinical studies have identified up-regulation of the critical antiapoptosis gene bcl-2 in MCC. We conducted a multicenter phase II trial of the novel bcl-2 antisense agent (G3139, Genasense) in patients with advanced MCC. METHODS: Twelve patients (9 men, 3 women) with histologically confirmed metastatic or regionally recurrent MCC were enrolled. Ten patients (83%) had received prior chemotherapy. Eight patients (67%) had Karnofsky performance status of 90 to 100. Patients received continuous IV infusion of G3139 (7 mg/kg/d) via central venous access in an outpatient setting for 14 days, followed by a 7-day rest period. Response was assessed at 6-week intervals. Patients were allowed to continue therapy until unacceptable toxicity or disease progression. RESULTS: No objective responses were observed. The best response was stable disease in 3 patients and progressive disease in 9 patients. A median of 4 doses per patient (total 46 doses) was administered. Dose delays and/or reductions were required in 6 patients. One patient developed grade 4 lymphopenia. One patient developed grade 3 renal failure characterized by grade 3 elevated creatinine and grade 4 hyperkalemia. Other grade 3 events included cytopenia (n = 5), aspartate aminotransferase/alanine aminotranferease elevation (n = 3), hypophosphatemia (n = 2), and pain (n = 1). The most frequent grade 1 to 2 toxicities were elevated creatinine, ALT elevation, hypokalemia, lymphopenia, and fatigue. CONCLUSIONS: Bcl-2 antisense therapy (G3139) was well tolerated among patients with advanced MCC. Although probable antitumor activity was documented in 1 patient, no objective responses per Response Evaluation Criteria in Solid Tumors criteria were observed. PMID- 19307958 TI - Phase II study of epirubicin in combination with weekly docetaxel for patients with advanced NSCLC who have failed or relapsed after the frontline platinum based chemotherapy. AB - BACKGROUND: We conducted a phase II study to evaluate the efficacy and toxicity of weekly docetaxel combined with epirubicin on D15 as second-line chemotherapy in Taiwanese patients with advanced non small cell lung cancer (NSCLC) who failed or relapsed after the frontline platinum-based chemotherapy. PATIENTS AND METHODS: Patients with histologically confirmed advanced NSCLC (Stage IIIB-IV) were entered into this Phase II trial. Eastern Cooperative Oncology Group performance status was 0 to 2 and adequate organ function was required. Docetaxel, 30 mg/m, was given intravenously on days 1, 8, and 15 for 30 minutes and epirubicin, 60 mg/m, was given intravenously on day 15, then following one week of rest. Treatment was repeated every 4 weeks for a maximal total of 6 cycles. RESULTS: Of the 43 eligible patients, 39 patients were evaluated for response, and all were evaluated for toxicity. The overall response rate was 11.6% [95% confidence interval (CI), 1.6-21.6%]. The median time to disease progression for all patients was 2.8 months (95% CI 1.3-4.3%). The median survival time for all patients was 7.7 months (95% CI 5.5-9.9%). The 1-year survival was 32.6% (95% CI 25.4%-39.7%). The major hematologic toxicities were neutropenia, 8/43 (19%) with grade 3-4 neutropenia, as well as anemia, 6/43 (14%) with grade 3-4 anemia. Nonhematological toxicities were modest. Fatigue was common, 77.8% in all, but only 3 (7%) patients with grade 3-4 toxicities. Diarrhea was also common but not severe, 7/43 (16%) with grade 1-2 episodes, and 1/43 (2%) with grade 3-4 episodes. Nail changes, peripheral edema, lacrimation, and alopecia were mild. Hepatic and renal impairment was also only mild. CONCLUSION: Combining weekly doses of docetaxel 30 mg/m with epirubicin 60 mg/m on D15 was not shown to improve both efficacy and tolerability for advanced NSCLC patients who have relapsed disease after frontline platinum-based chemotherapy. PMID- 19307959 TI - Stomal recurrence after total laryngectomy: a clinicopathological multivariate analysis. AB - OBJECTIVE: To evaluate the possible risk factors associated with recurrence of stomal recurrence after total laryngectomy that may be used as evidence of the efficacy of select preventive procedures. METHODS: Various clinicopathologic factors of 548 patients who underwent total laryngectomy for laryngeal cancer at our hospital between January 1995 and December 2004 were reviewed. Univariate and multivariate logistic regression models were constructed to evaluate the risk factors for odds ratio (OR) and statistical significance. RESULTS: Recurrence of stomal recurrence was observed in 48 cases and the median time to recurrence was 9.1 months. Univariate analysis showed that primary location (P = 0.000), T-stage (P = 0.000), preoperative tracheotomy (P = 0.001), and positive surgical margin (P = 0.000) were significant factors associated with stomal recurrence. Preoperative tracheotomy had no effect on the incidence of stomal recurrence in the T3 or T4 subgroups. Multivariate analysis showed that T-stage (OR = 1.687; P = 0.049), primary location (OR = 2.387; P = 0.012), and surgical margin (OR = 2.278; P = 0.020) were independent predictive factors for stomal recurrence. CONCLUSIONS: The prognosis of patients with stomal recurrence after laryngectomy is very poor. Identification of patients with high-risk factors is essential for offering more selective treatments to prevent their evolution. PMID- 19307960 TI - Adenosine receptor antagonists improve short-term object-recognition ability of spontaneously hypertensive rats: a rodent model of attention-deficit hyperactivity disorder. AB - The strain of spontaneously hypertensive rats (SHR) is considered a genetic model for the study of attention-deficit hyperactivity disorder (ADHD), as it displays hyperactivity, impulsivity and poorly sustained attention. Recently, we have shown the involvement of adenosinergic neuromodulation in the SHR's short-term and long-term memory impairments. In this study, we investigated the performance of male and female SHR in a modified version of the object-recognition task (using objects with different structural complexity) and compared them with Wistar rats, a widely used outbred rat strain for the investigation of learning processes. The suitability of the SHR strain to represent an animal model of ADHD, as far as mnemonic deficits are concerned, was pharmacologically validated by the administration of methylphenidate, the first-choice drug for the treatment of ADHD patients. The role of adenosine A1 and A2A receptors in object discrimination was investigated by the administration of caffeine (nonselective antagonist) or selective adenosine receptor antagonists. Wistar rats discriminated all the objects used (cube vs. pyramid; cube vs. T-shaped object), whereas SHR only discriminated the most structurally distinct pairs of objects (cube vs. pyramid). Pretraining administration of methylphenidate [2 mg/kg, intraperitoneal (i.p.)], caffeine (1-10 mg/kg, i.p.), the selective adenosine receptor antagonists DPCPX (8-cyclopenthyl-1,3-dipropylxanthine; A1 antagonist, 5 mg/kg, i.p.) and ZM241385 (A2A antagonist, 1.0 mg/kg, i.p.), or the association of ineffective doses of DPCPX (3 mg/kg) and ZM241385 (0.5 mg/kg), improved the performance of SHR in the object-recognition task. These findings show that the discriminative learning impairments of SHR can be attenuated by the blockade of either A1 or A2A adenosine receptors, suggesting that adenosinergic antagonists might represent potentially interesting drugs for the treatment of ADHD. PMID- 19307961 TI - Effects of abstinence or extinction on cocaine seeking as a function of withdrawal duration. AB - The resumption of drug-seeking behavior after abstinence or extinction is commonly studied model for relapse in addiction. For the benefits of extinction training over a given withdrawal period to be determined, it is necessary to discriminate between the potentially overlapping occurrence of incubation with that of spontaneous recovery. This comparison has been assessed using a between subjects design in groups of abstinent and extinguished rats tested at various withdrawal periods after cocaine self-administration. Multiple forms of priming were used to evoke the resumption of drug seeking, as different priming stimuli have been reported to use distinct neurobiological mechanisms and therefore may exhibit different temporal characteristics. In abstinent animals (30 days), neither the noncontingent conditioned stimuli-primed nor the noncontingent cocaine-primed drug seeking displayed incubation, whereas the drug seeking provoked by exposure to the contextual cues of the operant chamber significantly increased. In extinguished animals, evidence of spontaneous recovery of responding was observed after priming with exposure to either contextual or cocaine-priming stimuli. Finally, extinction training remained effective in reducing the reinstatement response levels after contextual or cocaine priming even if such training was initiated after an extended period (24 days) of abstinence. These findings provide further insight into the time-dependent effects of abstinence and extinction on the resumption of drug-seeking behavior. PMID- 19307962 TI - Surgeons' perspective on percutaneous valve repair. AB - The emergence of percutaneous therapy for valvular heart disease may provide valuable options for patients unable to tolerate conventional valve surgery. To optimize the results of percutaneous valve interventions, we must integrate the lessons learned from years of surgical experience. PMID- 19307964 TI - Sociological and ethical issues in transplant commercialism. AB - PURPOSE OF REVIEW: 'Global transplant commercialism' (practices and policies involving international trade in organs from living vendors, e.g., 'transplant tourism') is currently subjected to unprecedented criticism. In parallel, the debate around 'local transplant commercialism' (practices and policies that confine trade in organs from living vendors to national markets or economic unions) is heating up. In an attempt to assess the potential outcomes of these trends, this article reviews and discusses some sociological and ethical issues, ending with a proposal for a reinvigorated anticommercialist strategy. RECENT FINDINGS: The current international campaign against global transplant commercialism is conducted by an ad hoc alliance between strange bedfellows, proponents of local transplant commercialism on the one hand and opponents of any transplant commercialism on the other. Disparities in the rigor of the respective ethical discourses, the expanding list of precedents of legitimized commerce in the human body, and the political economy of transplantation, all suggest that the former have the upper hand. SUMMARY: Recent achievements in the struggle against international organ trafficking may not herald the abolition of transplant commercialism but rather presage its reconfiguration in deglobalized forms. In light of such a prospect, those who wish to prevent the pervasive commodification of the human body from entering the gates of transplant medicine should consider devising a new, perhaps more radical, strategy. PMID- 19307965 TI - Incentives for organ donation in the United States: feasible alternative or forthcoming apocalypse? AB - PURPOSE OF REVIEW: Several factors have generated interest in proposals to offer incentives in exchange for kidneys from living donors, including the growing shortage of available organs, the apparent asymptote of traditional means of organ procurement, and the intimate link between the inadequacies of organ procurement policies in developed countries with the flourishing of underground organ trafficking in developing countries. RECENT FINDINGS: Herein, we review the scope and dimensions of the growing shortage of organs in the United States, with attention to how each of the proposed solutions to same has proven insufficient. With special attention to the concerns leveled by Gabriel Danovitch in his 'Open Letter,' we conclude that each of his concerns are unfounded, and offer a prospectus on how a trial of such systems might be pursued in the United States. SUMMARY: The failure of current approaches to organ procurement in the United States and other developed countries has led to unnecessary suffering and death, with morally unacceptable consequences for developing countries. For these reasons, a structured trial of incentives for organ procurement in the United States is a moral imperative. PMID- 19307966 TI - Loin pain hematuria syndrome: a psychiatric and surgical conundrum. AB - PURPOSE OF REVIEW: The loin pain hematuria syndrome presents a dilemma with regards to the etiology, as well as the treatment of this rarely seen entity. In view of the increasing frequency of diagnosis, and the question of whether this disorder constitutes a somatoform disorder or a physical disorder remedied through renal autotransplantation we should familiarize ourselves with this condition, so as to clarify its nature. RECENT FINDINGS: There may be a subset of loin pain hematuria syndrome patients that have a somatoform disorder. Patients appear to have better outcomes with autotransplantation, than with intraureteric capsaicin treatment or renal denervation. SUMMARY: This paper attempts to provide an overview of the topic and propose further investigation to better determine whether a subset of these patients have a somatoform disorder. PMID- 19307967 TI - Current world literature. PMID- 19307968 TI - Psychological treatments for stimulant misuse, comparing and contrasting those for amphetamine dependence and those for cocaine dependence. AB - PURPOSE OF REVIEW: The aim is to compare and contrast psychological treatments for amphetamine and cocaine dependence. RECENT FINDINGS: Stimulant dependence, in the form of cocaine or amphetamine/methamphetamine dependence, is prevalent worldwide, and their ratio may vary across different countries and regions of countries. The treatment of stimulant disorders has greatly advanced in recent years, and scientific evaluation of behavioral therapies, using randomized clinical trials designs and a stage-wise approach, have demonstrated the safety and efficacy of interventions. Psychological interventions such as cognitive behavioral therapy and contingency management for cocaine and methamphetamines use disorders are well tolerated and moderately effective in achieving drug abstinence. There is evidence that contingency management interventions can help to improve retention in treatment and, in turn, other treatment outcomes. Although there are important differences in the neuropsychiatric and medical consequences of cocaine and amphetamine use disorders, there is currently no evidence for a differential treatment effect of any psychosocial treatment in the management of these disorders. SUMMARY: As there are no Food and Drug Administration-approved medications for the treatment of these disorders, psychological interventions form the basis of their treatment. More research is needed to address the specific psychosocial needs of cocaine and amphetamine dependent individuals in order to improve their treatment outcomes. PMID- 19307970 TI - The effective duration of analgesia after intrathecal morphine in patients without additional opioid analgesia: a randomized double-blind multicentre study on orthopaedic patients. AB - BACKGROUND AND OBJECTIVE: To know whether the application of patient-controlled analgesia devices could be avoided if intrathecal morphine is given in combination with spinal anaesthesia. METHODS: In a randomized, double-blind multicentre study, 188 orthopaedic patients were randomized to receive intrathecally placebo, 0.1 mg morphine or 0.2 mg morphine in addition to 15 mg bupivacaine. The primary outcome parameter was the number of patients without any additional request for opioid during a period of 72 h after surgery. RESULTS: Patients with 0.1 or 0.2 mg morphine showed a significant reduction in opioid requests compared with placebo during 72 h after surgery (P = 0.0001). At 24 h after surgery, the rate of patients who required additional opioid analgesia was 71% in the placebo group, 51% in the 0.1 mg morphine group and 31% of the patients in the 0.2 mg morphine group. After 0.2 mg morphine, systemic opioid requirements at 24 h were significantly lower than those in patients with 0.1 mg morphine (P < 0.05). Intrathecal morphine was not associated with an increased frequency of respiratory depression. Forty per cent of patients with intrathecal morphine did not ask for systemic opioids. CONCLUSION: Intrathecal morphine in a dose of 0.1 and 0.2 mg provides effective analgesia for up to 48 h without any need for systemic opioids at all in many patients. PMID- 19307971 TI - Soluble TREM-1 is not suitable for distinguishing between systemic inflammatory response syndrome and sepsis survivors and nonsurvivors in the early stage of acute inflammation. AB - BACKGROUND AND OBJECTIVE: To evaluate plasma levels of soluble TREM-1 (sTREM-1) in patients with systemic inflammatory response syndrome (SIRS), severe sepsis, and septic shock and to determine whether plasma sTREM-1 could be used as a diagnostic and prognostic marker in sepsis in the surgical ICU. METHODS: The study was designed as an observational noninterventional clinical study in a surgical ICU of a university hospital. For this, 65 intensive care patients were enrolled within the first 24 h after onset of SIRS (n = 11), severe sepsis (n = 39) or septic shock (n = 15). In addition, 21 healthy volunteers served as controls. At days 0, 1, and 3 after diagnosis, plasma sTREM-1 was measured by ELISA. RESULTS: Plasma sTREM-1 concentrations in healthy controls did not significantly differ from those in patients with SIRS, severe sepsis, or septic shock at days 0, 1, and 3. Survivors were defined as septic patients surviving for at least 28 days. There were no differences in plasma sTREM-1 levels between survivors (n = 22) and nonsurvivors (n = 27) on any day. CONCLUSIONS: In this study in patients with SIRS, severe sepsis, or septic shock, plasma sTREM-1 levels were not elevated as compared with healthy controls. Measurement of plasma sTREM-1 did not distinguish between patients with SIRS, severe sepsis, or septic shock or between survivors and nonsurvivors. The suggested role of sTREM-1 as a diagnostic and prognostic marker in sepsis should be carefully verified. PMID- 19307972 TI - No adjustment vs. adjustment formula as input weight for propofol target controlled infusion in morbidly obese patients. AB - BACKGROUND AND OBJECTIVE: The purpose of this prospective, randomized, double blind study was to determine the predictive performance of target-controlled infusions of propofol in morbidly obese patients using the 'Marsh' pharmacokinetic parameter set. METHODS: Twenty-four patients (ASA II or III, age 25-62 years, BMI 35.5-61.7) were randomly allocated to receive propofol target controlled infusion based on a weight adjustment formula (group adjusted) or without adjustment [group total body weight (TBW)]. Anaesthesia was induced by a propofol-targeted concentration of 6 microg ml that was subsequently adapted to maintain stable bispectral index values ranging between 40 and 50. Arterial blood samples were collected before the start of the infusion and every 15 min thereafter to determine the predictive performances. RESULTS: There were no statistically significant differences between the groups with regard to performance errors, divergence and wobble. Results are presented as median (interquartiles). Median performance error and median absolute performance error were -31.7 (-35.9, -19.4) and 31.7% (20.2, 35.9) for group adjusted and -16.3 ( 26.3, 2.2) and 20.6% (14.8, 26.9) for group TBW, respectively. Wobble median value was 7.4% (3.8, 8.4) for group adjusted and 8.2% (7.0, 9.6) for group TBW. As for wobble and divergence, no statistically significant differences were found between groups. CONCLUSION: Weight adjustment causes a clinically unacceptable performance bias, which is not corrected when TBW is used as an input to the 'Marsh' model. It is, therefore, advisable to administer propofol to morbidly obese patients by titration to targeted processed-EEG values. PMID- 19307973 TI - Effects of temperature gradient reduction in three different carbon dioxide absorbents. AB - BACKGROUND AND OBJECTIVE: Temperature gradients in CO2 absorbents may locally increase the water content by condensation. We hypothesized that temperature gradient reduction (TGR) would prevent increased water content, thus preserving the reactivity of the CO2 absorbent and thereby increasing its time to exhaustion (longevity). The purpose of this study was to compare the effects of TGR on the longevity of CO2 absorbent with three different types of CO2 absorbents. METHODS: We constructed a novel TGR canister. Experiments were conducted using three different types of CO2 absorbents: Dragersorb 800 Plus (D800), Dragersorb Free and Amsorb Plus. One kilogram of fresh CO2 absorbent of each type was placed into two types of canister: the conventional control canister (n = 6) and the TGR canister (n = 6). RESULTS: In the case of Dragersorb Free, the TGR canister most effectively and specifically prevented local increase in water content of the CO2 absorbent and markedly increased the longevity (30% increase) compared with the control canister. In the case of Amsorb Plus, the TGR canister also prevented local excessive water content, but the increase in longevity was smaller (17% increase). In the case of D800, the TGR canister markedly increased the longevity (27% increase), but its prevention of local excessive water content was smaller. CONCLUSIONS: TGR is a useful method to prevent local increase in water content and improve the longevity of CO2 absorbent. The effectiveness of TGR on longevity and water content changes varied in the different types of CO2 absorbent. PMID- 19307974 TI - Trainee anaesthetists' attitudes to error, safety and the law. AB - BACKGROUND AND OBJECTIVE: Anaesthetists are at increased risk of successful prosecution for manslaughter (nonculpable homicide) compared with other clinicians. METHODS: This questionnaire study assessed the attitudes of 109 British trainee anaesthetists (response rate 87%) to medical error, patient safety and the law relating to anaesthesia. RESULTS: Fifty-seven per cent had made an error that caused harm to patient. When asked to consider the worst error that they had made, 81% of 86 respondents had reported the error, but only 31% of reports were investigated further; 68% had informed the patient of their error. Medication (47%) and procedural errors (37%) were commonest. 'Best practice' guidance was followed in only 38% of instances. Media exposure (92%) was considered the likeliest reason for the increase in manslaughter prosecutions. Knowledge of the law relevant to anaesthesia was poor. Trainees had received limited training in decreasing their exposure to the legal process. CONCLUSION: In order to reduce their legal liability and improve patient safety, junior anaesthetists must practise according to professional guidelines and seek professional training in medicolegal matters. PMID- 19307975 TI - Angiotensin-converting enzyme insertion/deletion polymorphism in patients with acute and chronic pancreatitis. AB - BACKGROUND: Reduction in angiotensin-converting enzyme (ACE) activity has been shown to attenuate pancreatic stellate cell activation and pancreatic fibrosis and suggested as a potential treatment for chronic pancreatitis. The ACE gene insertion/deletion (I/D) polymorphism in intron 16 accounts for nearly half the variation in serum ACE levels. This study determined the frequency of the I/D polymorphism in patients with acute and chronic pancreatitis. METHODS: In total, 887 patients (346 with alcoholic, 443 with nonalcoholic, and 98 with acute pancreatitis) were enrolled, and were compared with 1294 healthy controls. Genotyping of the I/D polymorphism was performed by PCR or melting curve analyses. RESULTS: No significant differences were found in the prevalence of the ACE-deletion genotype frequencies when patients with alcoholic (27.5%), nonalcoholic (26.4%), and acute pancreatitis (32.7%) were compared with controls (26.9%). Likewise, allele frequencies of the ACE deletion polymorphism were not significantly different in patients with alcoholic (53.8%), nonalcoholic (50.6%), and acute pancreatitis (54.1%) and controls (52.7%). CONCLUSION: The I/D polymorphism of the ACE gene was not found to be associated with acute and chronic pancreatitis. PMID- 19307976 TI - Overexpression of liver-specific cytochrome P4502E1 impairs hepatic insulin signaling in a transgenic mouse model of nonalcoholic fatty liver disease. AB - OBJECTIVE: Cytochrome P4502E1 (CYP2E1) expression in the liver is increased in nonalcoholic fatty liver disease. The aim of this study was to determine whether CYP2E1 overexpression in the liver interferes with insulin signaling pathways in a mouse model of nonalcoholic fatty liver disease. METHODS: Male mice containing the human CYP2E1 transgene under control of the mouse albumin enhancer-promoter (Tg) and control, nontransgenic mice were fed a diet containing 20% calories from fat for 8 months ad libitum. MEASUREMENTS: Liver injury was measured by histology and alanine aminotransferase. Malondialdehyde and protein carbonyls were measured as markers of oxidative stress. Total and phosphorylated proteins involved in the insulin signaling cascade were measured by western blotting. RESULTS: Tg mice had higher fasting insulin, and greater hepatic fat accumulation and histological liver injury. Malondialdehyde and protein carbonyls were increased in Tg mice liver indicating increased oxidative stress. Tyrosine phosphorylation of insulin receptor substrates 1 and 2, and serine phosphorylation of PKB/Akt, were significantly decreased in Tg mice. Serine phosphorylation of glycogen synthase kinase 3alpha was decreased in Tg mice and liver glycogen content was decreased correspondingly. Serine phosphorylation of the transcription factor Fox01a was decreased, and expression of the enzyme phosphoenolcarboxykinase was increased in Tg mice. CONCLUSION: Hepatocyte-specific overexpression of CYP2E1 increased hepatic oxidative stress in the liver, fasting insulin, and histological liver damage. CYP2E1 overexpression reduced hepatic insulin signaling and reduced glycogen storage and increased glucose synthesis. Overall, this study suggests an association of hepatic CYP2E1 with increased oxidative stress, increased systemic insulin resistance, decreased insulin signaling in the liver and increased hepatic fat accumulation. PMID- 19307977 TI - Increased number of methylated CpG islands correlates with Helicobacter pylori infection, histological and serological severity of chronic gastritis. AB - BACKGROUND: Promoter hypermethylation of tumor suppressor genes is one of the major events in gastric carcinogenesis. Promoter hypermethylation is also present in non-neoplastic gastric epithelium as age-related phenomenon and some reports suggest the potential association between promoter hypermethylation and Helicobacter pylori infection. Here, we examined whether methylation of multiple promoter CpG islands would occur by H. pylori infection and correlate with histological or serological severity of chronic gastritis. METHODS: One hundred and ninety-one gastric mucosa samples were obtained by endoscopy. The promoter methylation status of the p14, p16, DAP-kinase and CDH1 genes were determined by methylation-specific-polymerase chain reaction. The degree of gastritis in the antrum was assessed according to the updated Sydney system in 150 participants. The pepsinogen (PG) I/II ratio was calculated based on the data of serum PG I and PG II levels measured by radioimmunoassay in 54 selected cases. RESULTS: CpG island methylation was found in 32.5% for p14, 35.1% for p16, 43.5% for DAP kinase and 36.1% for CDH1, whereas non, 1, 2, 3, and all methylation of four promoter CpG sites were present in 46 (24.1%), 59 (30.9%), 46 (24.1%), 30 (15.7%), and 10 (5.2%) participants, respectively. A strong association between the increased number of methylated CpG islands and H. pylori infection was observed (P<0.0001). An increased number of methylated CpG islands was also associated with severity of neutrophil infiltration (P<0.0001), mononuclear cell infiltration (P<0.0001) and atrophy (P=0.0021) in all, and severity of neutrophil infiltration (P=0.0177) and mononuclear cell infiltration (P=0.0004) in H. pylori positive participants. An increased number of methylated CpG islands correlated with lower PG I/II ratio in all (P=0.0105) and H. pylori-infected participants (P=0.074). CONCLUSION: Multiple promoter CpG islands would be methylated by H. pylori infection, and an increased number of methylated CpG sites correlate with histological and serological severity of chronic gastritis. PMID- 19307978 TI - Screening for familial colorectal cancer with a sensitive immunochemical fecal occult blood test: a pilot study. AB - OBJECTIVE: Colonoscopy is empirically recommended as the first choice screening strategy in first-degree relatives of patients with colorectal cancer (CRC). However, this strategy is accepted by less than 40% of the risk population and two-thirds of screened individuals and renders a normal exploration. This pilot study assessed the accuracy of a latex agglutination immunochemical fecal occult blood test (LA-FOBT) for detecting advanced colorectal neoplasm (cancer or adenomatous polyps > or =1 cm in size, with villous pattern or high grade dysplasia) in asymptomatic first-degree relatives of patients with CRC. METHODS: One hundred and sixty-nine first-degree relatives of 135 index cases were prospectively included. All participants received a sensitive LA-FOBT (hemoglobin detection limit of 50 ng/ml buffer), and were invited to undergo colonoscopy. On the whole, 116 (69%) participants returned LA-FOBT and underwent colonoscopy. RESULTS: LA-FOBT was positive in 19 of 116 (16%) cases. Colonoscopy detected neoplasms in 49 of 116 (42%) patients: 37 of 116 (32%) were nonadvanced adenomas and 12 of 116 (10%) advanced adenomas. LA-FOBT detected 10 of 12 (83%) advanced adenomas showing a sensitivity, specificity, positive predictive value, and negative predictive value of 83, 91, 53, and 98%, respectively. In patients with positive LA-FOBT, 1.9 colonoscopies were necessary for detecting one advanced adenoma, whereas in case of not performing this test 10 colonoscopies would be needed. Overall, approximately 80% of screening colonoscopies could be precluded using a LA-FOBT. CONCLUSION: One-time screening with LA-FOBT successfully detects advanced colorectal adenomas and may save unnecessary colonoscopies in first degree relatives of patients with CRC. PMID- 19307979 TI - Left main coronary artery aneurysm in young patient with acute myocardial infarction. AB - Aneurysms of the left main coronary artery (LMCAA) are extremely uncommon, with an incidence of 0.1%. The main etiologic factor is atherosclerosis. Other causes include connective tissue disorders, trauma, vasculitis, congenital, mycotic, and idiopathic. These dilated sections of coronary artery are not benign pathology because they are subject to spasm, thrombosis, and subsequent distal embolism, spontaneous dissection and rupture. Treatment options include anticoagulation, custom-made covered stents, reconstruction, resection and exclusion with bypass. Our report on a young case illustrates the potential complication of LMCAA and presents its management. PMID- 19307980 TI - Electromechanical mapping of the left ventricle: possible tool for online decision making in the catheterization laboratory. AB - Clinical decision making in intervention cardiology often depends on information about the presence of myocardial viability and the extent of ischemia. Especially in the case of an occluded collaterally filled coronary branch, online decision making in selected patients may accelerate and improve patient care. The electromechanical NOGA mapping system offers the opportunity for online viability assessment. We describe two cases in which this diagnostic tool was used during daily practice. In our opinion, NOGA mapping can be helpful for 'online' viability evaluation in patients with an occluded collaterally filled coronary artery. In these patients, noninvasive viability evaluation may cause unnecessary delay in the overall treatment approach. PMID- 19307981 TI - Utility of bone densitometry in diagnostic evaluation and monitoring in regional migratory osteoporosis. AB - Regional migratory osteoporosis (RMO) is a rare, self-limited disease of unknown etiology, characterized by migratory arthralgia of the weight-bearing joints of the lower extremities. RMO is most commonly seen in middle-aged adults, particularly men. Several case reports suggest that bone changes seen in RMO may be detected as focal decreases in bone mineralization, but the utility of serial bone density studies has yet to be formally evaluated in this syndrome. We describe a case of RMO with dramatic focal changes in bone mineral density that corresponded to the underlying clinical course. Serial bone density testing clearly documented significant bone loss and subsequent recovery of bone density in areas affected by RMO. Our data suggest that sequential densitometric examination of affected areas may be useful to confirm the diagnosis of RMO, and for subsequent monitoring of bone loss and therapeutic response. PMID- 19307982 TI - Long-term decline in renal function is linked to initial pulse pressure in the essential hypertensive. AB - OBJECTIVES: In the absence of malignant hypertension, the mechanisms for the decline in renal function in hypertensive patients are not well known. Several recent studies, essentially cross-sectional, point to a role for an increase in arterial stiffness and its corollary, the increased pulse pressure (PP), in barotrauma of the renal glomerulus. METHODS: We examined relations between the PP measured on consultation or by 24-h ambulatory blood pressure monitoring and the long-term decline in renal function in a population of essential hypertensive patients initially untreated, with normal renal function and without proteinuria. We evaluated the renal outcome of 375 patients of mean age 49 years in a baseline state over a mean follow-up period of 14 years. RESULTS: At follow up, the glomerular filtration rate estimated from the modification of diet in renal disease formula was below 60 ml/min per 1.73 m2 in 51 of these patients, two of whom required dialysis. The blood pressure parameter best correlated with subsequent renal failure, independently of other factors of risk such as age or type 2 diabetes, was the PP measured either in consultation or by ambulatory blood pressure monitoring before starting treatment. CONCLUSION: The PP either measured on consultation or by ambulatory monitoring emerged as an independent determinant of the decline in renal functions in this population, pointing to the possibility of barotrauma of the glomeruli from increased arterial stiffness. PMID- 19307983 TI - Endothelial damage due to impaired nitric oxide bioavailability triggers cerebral aneurysm formation in female rats. AB - OBJECTIVE: Epidemiological data indicate a high incidence of cerebral aneurysms in postmenopausal women. To elucidate the pathogenesis of cerebral aneurysms, we focused on the contribution of endothelial damage in rats. METHODS: We induced estradiol deficiency by oophorectomy (OVX), hypertension, or both, and hemodynamic stress in 7-week-old female Sprague-Dawley rats. They were then given hormone-replacement therapy with 17beta-estradiol or an angiotensin II type 1 receptor blocker (ARB). The effects of estradiol, angiotensin II type 1 receptor blocker, or both on cultured endothelial cells were also examined. RESULTS: The number of anomalously shaped endothelial cells was higher in OVX than hypertensive rats (P < 0.05). Rats subjected to hypertension and OVX exhibited a marked increase in the incidence of saccular cerebral aneurysms. Estradiol or angiotensin II type 1 receptor blocker treatment reduced this incidence (P < 0.05). The endothelial nitric oxide synthase (eNOS) mRNA level in the intracranial artery of OVX and hypertensive and OVX rats was low (P < 0.05). Immunohistochemically, the expression of eNOS and estrogen receptor alpha (ERalpha) in the vascular wall of hypertensive and OVX rats was decreased; angiotensin II and the nicotinamide adenine dinucleotide phosphate oxidase subunits nicotinamide adenine dinucleotide phosphate oxidase 4 and p22phox were strongly expressed in cerebral aneurysms. In the absence of estradiol, eNOS was downregulated and nicotinamide adenine dinucleotide phosphate oxidase expression was increased in endothelial cells; angiotensin II augmented these phenomena. The regulation of eNOS was mediated by ERalpha. These results suggest that estrogen deficiency induces endothelial dysfunction and reactive oxygen species generation, triggering endothelial damage that leads to cerebral aneurysms and that hypertension is an additional risk factor. CONCLUSION: A therapy targeted at the endothelium and management of hypertension may help to prevent cerebral aneurysms. PMID- 19307984 TI - Family-based analysis of apelin and AGTRL1 gene polymorphisms with hypertension in Han Chinese. AB - OBJECTIVE: Apelin and AGTRL1 (angiotensin receptor-like 1), elements of a newly identified pathway with a role in counter regulating the renin-angiotensin system, have been implicated in blood pressure regulation. This study aims to assess whether the apelin and AGTRL1 genetic polymorphisms might contribute to essential hypertension or its related phenotypes. METHODS: We recruited 1015 Han Chinese from 248 families with essential hypertension. Each individual was genotyped for 6 single nucleotide polymorphisms (SNPs) in apelin and 6 SNPs in AGTRL1. Data were analyzed using the family-based association test (FBAT) and the haplotype-based association test (HBAT). RESULTS: FBAT analysis showed that two SNPs rs3761581 and T-1860C within apelin conferred significant association with hypertension and its related phenotypes even after correcting for age and gender. Three SNPs (rs7119375, rs10501367 and rs9943582) within AGTRL1 were found to be associated with hypertension, BMI and the onset age of hypertension, whereas after correction, only marginal associations were noted. Of the common haplotypes (with frequencies over 3%), haplotypes (A-T) and (C-C) comprising rs3761581 and T 1860C in apelin and haplotype (G-G) comprising rs10501367 and rs7119375 in AGTRL1 were shown to be significantly associated with hypertension, BMI and the onset age of hypertension, even after a permutation correction. CONCLUSIONS: Our study suggests that genetic variation within apelin and AGTRL1 likely contributes to essential hypertension, BMI and the onset age of hypertension. Future well designed epidemiological or functional studies would be warranted to validate this hypothesis. PMID- 19307985 TI - Early lipopolysaccharide-induced reactive oxygen species production evokes necrotic cell death in human umbilical vein endothelial cells. AB - BACKGROUND: Endothelial dysfunction is a crucial step in the pathogenesis of cardiovascular diseases. Reactive oxygen species (ROS) generated in response to lipopolysaccharide (LPS) during sepsis promotes progressive endothelial failure. Typically, LPS-stimulated leukocytes produce pro-inflammatory cytokines, which trigger endothelial ROS production through NAD(P)H oxidase (Nox) activation, in a process that takes hours. Noteworthy, endothelial cells exposed to LPS may also generate ROS in just a few minutes. However, the mechanisms underlying this early event and its deleterious effect in endothelial function are unknown. Here, we investigated the mechanisms of early LPS-induced ROS generation and its effect in endothelial cell viability. METHODS: Human umbilical vein endothelial cells were exposed to LPS for 1-40 min to study ROS generation, cytokines expression, and signaling transduction by confocal microscopy, real-time PCR (RT-PCR), western blot, and immunoprecipation. Fourty-eight hour treatments were used to determine cell death by MTT assay, cell counting, and flow cytometry. Contribution of specific Nox isoform was evaluated using a siRNAs approach. RESULTS: LPS rapidly evoked a cytokine-independent ROS production, eliciting a rapid increase in p47phox phosphorylation by a phospholipase C/conventional protein kinase C and PI3-K signaling. It is noteworthy that the early LPS-induced ROS production triggered significant endothelial necrosis, which was prevented by a previous, but not a posterior, antioxidant treatment. The early LPS-induced ROS production as well as endothelial necrosis was totally dependent of Nox2 and Nox4 activity. CONCLUSION: Endothelial cells exposure to LPS triggers an early ROS production. Remarkably, this single early ROS production is enough to generate extensive endothelial cell death by necrosis dependent on the activity of Nox2 and Nox4. Because, in sepsis, ROS production can cause endothelial dysfunction, results here provided may be relevant when considering the development of strategies for sepsis therapy. PMID- 19307986 TI - Altered nitric oxide calcium responsiveness of aortic smooth muscle cells in spontaneously hypertensive rats depends on low expression of cyclic guanosine monophosphate-dependent protein kinase type I. AB - OBJECTIVES: The nitric oxide/cyclic guanosine monophosphate (GMP)/cyclic GMP dependent protein kinase type I (cGKI) pathway has been extensively investigated in the spontaneously hypertensive rat (SHR) as a possible pathogenetic factor. Therefore, we investigated the role of nitric oxide/cGKI on intracellular calcium dynamics ([Ca2+]i) of aortic smooth muscle cells isolated from control normotensive Wistar Kyoto rats (WKY) and SHR. METHODS: Rat aortic smooth muscle cells (RASMCs) were obtained from 12 to 16-week-old WKY and SHR. [Ca2+]i dynamics were monitored by imaging analysis of fura-2-loaded RASMCs. cGKI mRNA and cGKI protein expression were evaluated by reverse transcription-PCR and western blot. Plasmids codifying for enhanced green fluorescent protein (EGFP) or cGKIalpha EGFP were transfected on SHR RASMCs. RESULTS: Angiotensin II similarly increased [Ca2+]i in WKY and SHR RASMCs. In WKY RASMCs, S-nitroso-N-acetyl-DL-penicillamine (SNAP, 1-100 micromol/l) reduced the decay time of angiotensin II-induced [Ca2+]i transient. On the contrary, in SHR cells, SNAP was ineffective. Dibutyryl cyclic GMP (1-100 nmol/l), a membrane-permeable cyclic GMP analogue, behaved similarly to SNAP. In naive SHR RASMCs, cGKI mRNA and cGKI protein were low or absent. After transfection of a plasmid encoding for cGKIalpha-EGFP, the [Ca2+]i dynamic of SHR-transfected cells regained sensitivity to the nitric oxide/cyclic GMP pathway. CONCLUSION: The low expression of cGKI determines the lack of nitric oxide/cyclic GMP-dependent regulation on [Ca2+]i transient in SHR RASMCs. This alteration may contribute to the development of hypertension and explain suboptimal responses to nitroglycerin and other nitric oxide-releasing molecules in patients. PMID- 19307987 TI - Medication adherence in the MTA: saliva methylphenidate samples versus parent report and mediating effect of concomitant behavioral treatment. AB - OBJECTIVE: Although research supports the use of appropriately administered stimulant medication to treat children with ADHD, poor adherence and early termination undermine the efficacy of this treatment in real-world settings. Moreover, adherence measures often rely on parent report of medication use, and their validity and reliability are unknown. METHOD: Drawing on data from 254 participants in the NIMH Collaborative Multisite Multimodal Treatment Study of Children With Attention-Deficit/Hyperactivity Disorder, we examine the discrepancy between parents' verbal reports of medication adherence and physiological adherence measures determined via methylphenidate saliva assays collected at four time points during the 14-month treatment period. In addition, we examine the impact of physiologically documented medication adherence on parent- and teacher-reported outcomes through 14 months. RESULTS: Overall, nearly one fourth (24.5%) of the saliva samples indicated nonadherence. Among subjects, 63 (24.8%) of the 254 participants were nonadherent on 50% or more of their repeated saliva assays. Only 136 (53.5%) of the subjects were adherent at every time point at which saliva assays were taken, indicating that some degree of nonadherence characterized nearly half of all other NIMH Collaborative Multisite Multimodal Treatment Study of Children With Attention-Deficit/Hyperactivity Disorder-treated children. Findings also indicated that nonadherence produced greater deleterious effects in children in the medication-only condition compared with those receiving both medication and behavioral treatment. CONCLUSIONS: Same day saliva methylphenidate assays suggest that nearly half of the parents are inaccurate informants of their child's ADHD medication adherence and that parents may overestimate actual (physiological) adherence. This finding suggests the need for interventions to improve accuracy of parental report. Clinicians need to focus on adherence enhancement strategies to improve outcomes of children being treated with medication, particularly when benefits are suboptimal. PMID- 19307988 TI - Suppressive influences in the immune response to cancer. AB - Although much evidence has been gathered demonstrating that immune effectors can play a significant role in controlling tumor growth under natural conditions or in response to therapeutic manipulation, it is clear that malignant cells do evade immune surveillance in most cases. Considering that anticancer active specific immunotherapy seems to have reached a plateau of results and that currently no vaccination regimen is indicated as a standard anticancer therapy, the dissection of the molecular events underlying tumor immune escape is the necessary condition to make anticancer vaccines a therapeutic weapon effective enough to be implemented in the routine clinical setting. Recent years have witnessed significant advances in our understanding of the molecular mechanisms underlying tumor immune escape. These mechanistic insights are fostering the development of rationally designed therapeutics aimed to revert the immunosuppressive circuits that undermine an effective antitumor immune response. In this review, the best characterized mechanisms that allow cancer cells to evade immune surveillance are overviewed and the most debated controversies constellating this complex field are highlighted. PMID- 19307989 TI - Tumor-derived TGF-beta mediates conversion of CD4+Foxp3+ regulatory T cells in a murine model of pancreas cancer. AB - CD4+25+Foxp3+ regulatory T cells (Treg) play a critical role in the induction of tolerance to tumor-associated antigens and suppression of antitumor immunity. How Treg are induced in cancer is poorly understood. We reported previously that Treg are significantly elevated in the peripheral blood of patients with pancreas cancer and that in a murine pancreas cancer model induction of Treg seems to be transforming growth factor (TGF)-beta dependent. Here we provide additional evidence that Treg are increased locally within the tumor microenvironment by a mechanism that seems dependent on TGF-beta receptor expression and the presence of tumor derived TGF-beta. The murine pancreas cancer cell line Pan02 produces high levels of TGF-beta both in vitro and in vivo. In contrast, the esophageal murine cancer cell line, Eso2, does not. Immunohistochemical staining of Foxp3 in explanted tumors shows an identifiable population of Treg in the Pan02 (TGF-beta positive) tumors but not Eso2 (TGF-beta negative). Naive CD4+25-Foxp3- T cells, when adoptively transferred into Rag-/- mice, are converted into Foxp3+ Treg in the presence of Pan02 but not Eso2 tumors. Induction of Treg in Pan02 mice is blocked by systemic injection of an anti-TGF-beta antibody. If Rag-/- mice are instead reconstituted with naive CD4+25- T cells expressing a mutated TGF-beta receptor, induction of Foxp3+ Treg in Pan02 bearing mice is blocked. Collectively, these observations further support the role of TGF-beta in the induction of Treg in pancreas adenocarcinoma. PMID- 19307990 TI - The immune tolerance of cancer is mediated by IDO that is inhibited by COX-2 inhibitors through regulatory T cells. AB - Prostaglandin (PGE2), synthesized by cyclooxygenase-2 (COX-2), is associated with cellular immune tolerance during the process of cancer development. Induction of tolerance requires a specific environment in which dendritic cells and regulatory T cells (Tregs) play an essential role. It was recently shown that maturation of dendritic cells in the presence of indoleamine 2, 3-dioxygenase (IDO) results in activation of Tregs, and inhibition of COX-2 activity regulated IDO expression within the tumor microenvironment. Thus, we hypothesized that the tumor immune tolerance would be inhibited by COX-2 inhibitor and this inhibition would be mediated by IDO-dependent Tregs inhibition. The PGE2 in Lewis lung cancer cells (3LL) and serum of mice were measured for the evaluation of COX-2 inhibitors' local and systemic effects. The production of PGE2 in 3LL cells and serum of 3LL tumor-bearing mice were decreased by COX-2 inhibition. However, there were no significant differences in serum PGE2 levels among normal control and celecoxib treated nontumor-bearing mice. The accumulation of Tregs was reduced in the celecoxib-treated 3LL tumor-bearing mice. In addition, the expressions of COX-2, IDO, and Foxp3 were reduced in the mice treated with a COX-2 inhibitor, and this was found to correlate with a reduction in the size of tumor mass and metastasis. These results suggest that the antitumor effects of COX-2 inhibitors seemed to be correlated with the inhibition of IDO and Tregs. Therefore, COX-2 inhibitors might provide a therapeutic strategy for Tregs-induced tumor immune tolerance. PMID- 19307991 TI - CD40-activated apoptotic tumor cell-pulsed dendritic cell could potentially elicit antitumor immune response: involvement of up-regulation of B7-H3 expression. AB - Dendritic cells (DCs) initiate and direct immune responses. Previous in vitro and in vivo studies have showed that DCs matured with CD40 linking signal could potentially elicit and boost antitumor immunity, however, its molecular mechanism remain elusive. This study demonstrates that expression of B7-H3 on apoptotic cell-loading DCs is up-regulated markedly by CD40 activation but not by tumor necrosis factor-alpha stimulation. There was no significant difference found with CD40, CD80, or CD86 expressions when activated by CD40 or tumor necrosis factor alpha stimulation. In tumor-bearing mice, T cells conditioned with B7-H3-blocked on CD40-activated apoptotic tumor cell-pulsed DCs had a decreased ability to inhibit tumor growth. Therefore, it is hypothesized that high levels of B7-H3 expression contributes to the ability of CD40-activated tumor associated DCs in eliciting efficient antitumor immune response, given this fact the potentially significant clinical implications, CD40-activated DCs merit further considerations when preparing DCs for clinical application. PMID- 19307992 TI - Insertion of a targeting peptide on capsid surface loops of human papillomavirus type-16 virus-like particles mediate elimination of anti-dsDNA Abs-producing B cells with high efficiency. AB - The purpose of this study was to design chimeric human papillomavirus type-16 L1 virus-like particles (VLPs) and to explore the potential capacity of elimination to anti-dsDNA antibody-producing B cells. To test it, VLPs were achieved by combination of human papillomavirus type-16 L1 proteins inserted into a targeting peptide (DWEYSVWLSN) and plasmids encoding diphtheria toxin A ligand. Additionally, VLPs were cocultured with target cells to assess the killing efficiency by lactate dehydrogenase assay in vitro. Lastly, lupus-prone (BWF1) mice vaccinated with VLPs were used as a model to assess the killing efficiency in vivo. The results showed that the VLPs were constructed successfully, and possessed the potential of killing anti-dsDNA antibody-producing B cells with high efficiency. The findings indicate the possibility that the VLPs ablate autoreactive B cells represents a novel strategy in the immunotherapy of autoantibody-mediated diseases. PMID- 19307993 TI - Neem leaf glycoprotein induces perforin-mediated tumor cell killing by T and NK cells through differential regulation of IFNgamma signaling. AB - We have demonstrated augmentation of the CD3-CD56+ natural killer (NK) and CD8+CD56_ T-cell-mediated tumor cell cytotoxicity by neem leaf glycoprotein (NLGP). These NK and T cells were isolated from the peripheral blood of head and neck squamous cell carcinoma patients with a state of immunosuppression. NLGP induces TCRalphabeta-associated cytotoxic T lymphocyte (CTL) reaction to kill oral cancer (KB) cells. This CTL reaction is assisted by NLGP-mediated up regulation of CD28 on T cells and HLA-ABC, CD80/86 on monocytes. CTL-mediated killing of KB cells and NK-cell-mediated killing of K562 (erythroleukemic) cells are associated with activation of these cells by NLGP. This activation is evidenced by increased expression of early activation marker CD69 with altered expression of CD45RO/CD45RA. NLGP is a strong inducer of IFNgamma from both T and NK cells; however, IFNgamma regulates the T-cell-mediated cytotoxicity only without affecting NK-cell-mediated one. Reason of this differential regulation may lie within up-regulated expression of IFNgamma-receptor on T-cell surface, not on NK cells. This NLGP-induced cytotoxicity is dependent on up-regulated perforin/granzyme B expression in killer cells, which is again IFNgamma dependent in T cells and independent in NK cells. Although, FasL expression is increased by NLGP, it may not be truly linked with the cytotoxic functions, as brefeldin A could not block such NLGP-mediated cytotoxicity, like, concanamycin A, a perforin inhibitor. On the basis of these results, we conclude that NLGP might be effective to recover the suppressed cytotoxic functions of NK and T cells from head and neck squamous cell carcinoma patients. PMID- 19307994 TI - The Immune adjuvant properties of front-line carboplatin-paclitaxel: a randomized phase 2 study of alternative schedules of intravenous oregovomab chemoimmunotherapy in advanced ovarian cancer. AB - Oregovomab is a monoclonal antibody that recognizes CA125 and forms circulating immune complexes that can elicit immunity against both tumor antigen and tumor. This study was designed to assess combining this immunotherapy at 2 dosing schedules with front-line chemotherapy in patients with advanced ovarian cancer. Forty patients with stage III/IV carcinomas were randomized to receive a 2 mg oregovomab infusion either the same day [simultaneous infusion (SIM)] or 1 week after [1-week delayed (OWD)] standard carboplatin-paclitaxel chemotherapy at cycles 1, 3, and 5, then quarterly for up to 11 antibody doses. The primary end point was antibody response to oregovomab. Secondary end points included cellular immune response, response rate to front-line treatment, and progression-free survival. A different immune response pattern was observed between the SIM arm and the OWD arm, baseline plasma cytokines were balanced. Humoral immunity occurred more rapidly (P=0.0033) and with greater magnitude in the SIM arm. Absolute lymphocyte counts decreased in the SIM arm at cycles 3 and 5 compared with baseline. Treatment emergent CA125-specific cellular immunity was measured more commonly with SIM (P=0.04) and clinical parameters directionally favored this schedule. The immune responses were stronger than those measured in a previous maintenance monoimmunotherapy protocol. Immunotherapy-associated toxicity was minimal in this study. Front-line chemotherapy with carboplatin paclitaxel has immune adjuvant properties when combined with oregovomab immunotherapy; however, schedule is important. SIM strategies of carboplatin and paclitaxel should be further studied with oregovomab and other antigen-specific cancer immunotherapy approaches. PMID- 19307995 TI - Clinical grade OK432-activated dendritic cells: in vitro characterization and tracking during intralymphatic delivery. AB - Dendritic cells (DC) are under intense preclinical and early clinical evaluation for the immunotherapy of cancer. However, the optimal culture conditions and route of delivery for DC vaccination have not been established. Here we describe the first human application of DC matured with the bacterial agent OK432 (OK-DC), using a short-term serum-free culture protocol, which generates mature DC from CD14+ precursors after 5 days. These cells were prepared within the framework of a National Blood Service facility, demonstrating that DC represent a product which is potentially deliverable alongside current standardized cell therapies within the UK National Health Service. In vitro analysis confirmed that OK-DC were mature, secreted tumor necrosis factor-alpha, interleukin-6, and interleukin 12, and stimulated both T cell and natural killer cell function. To explore effective delivery of OK-DC to lymph nodes, we performed an initial clinical tracking study of radioactively labeled, unpulsed OK-DC after intralymphatic injection into the dorsum of the foot. We showed that injected DC rapidly localized to ipsilateral pelvic lymph nodes, but did not disseminate to more distant nodes over a 48-hour period. There was no significant toxicity associated with OK-DC delivery. These results show that OK-DC are suitable for clinical use, and that intralymphatic delivery is feasible for localizing cells to sites where optimal priming of innate and adaptive antitumor immunity is likely to occur. PMID- 19307996 TI - A phase 1 study of granulocyte macrophage colony-stimulating factor (sargramostim) and escalating doses of thalidomide in patients with high-risk malignant melanoma. AB - This phase 1 study evaluated the safety and tolerability of adjuvant treatment with subcutaneous granulocyte macrophage colony-stimulating factor (GM-CSF) administered in combination with escalating doses of thalidomide in patients with surgically resected stage II (T4), III, or IV melanoma at high risk for recurrence. Adjuvant treatment included GM-CSF 125 microg/m2 subcutaneously for 14 days and thalidomide at an initial dose of 50 mg/d, escalated in cohorts of 3 to 6 patients each to a maximum of 400 mg/d followed by 14 days of rest. Treatment was continued for up to 1 year in the absence of disease progression. Of 19 patients treated, the most common toxicities were grade 1/2 constipation (68%), fatigue (58%), neuropathy (42%), bone and joint pain (37%), and dyspnea, dizziness, injection site skin reaction, and somnolence (32% each). Thrombotic events in 3 of 19 patients (16%), including 1 treatment-related death, were the most serious adverse events and were thought to be due to thalidomide. With a median follow-up of 945 days (2.6 y), 8 (42%) patients were alive, including 1 with disease and 7 without evidence of disease. GM-CSF plus thalidomide as adjuvant therapy for patients with resected high-risk melanoma was associated with a high incidence of thrombotic events. Because life-threatening events are unacceptable in the adjuvant setting, up-front antithrombotic prophylaxis will be necessary for further evaluation of GM-CSF plus thalidomide as a viable regimen in this patient group. PMID- 19307997 TI - Combined cryoablation and GM-CSF treatment for metastatic hormone refractory prostate cancer. AB - A total of 12 patients with metastatic hormone refractory prostate cancer were treated by combining cryoablation and granulocyte macrophage colony-stimulating factor administration. Besides prostate-specific antigen (PSA) measurements, peripheral blood mononuclear cells were also obtained; the frequency of tumor specific T cells was tested ex vivo in an interferon-gamma enzyme-linked immunospot assay after stimulating with autologous prostate cancer-derived protein lysates. To assess cytolytic activity, T cells were coincubated with human prostate cancer cells (LNCaP) or renal cancer cells (GRC-1), and release of cytosolic adenylate kinase was measured by a luciferase assay. The median PSA decline percentage was 69.4% (range: 30.5% to 92.5%) and the median time to the nadir PSA was 4 months after therapy (range: 3 to 6). The median time to disease progress was 18 months, and 1 patient obtained a 92.5% PSA decline and a greater than 50% reduction of lung disease and survived 31 months. Four or 8 weeks after treatment, the tumor-specific T-cell responses were increased in peripheral blood mononuclear cell. The cytolytic activity against LNCaP was also increased significantly whereas no response was found against GRC-1. It seemed that there was no direct correlation between the degree of T-cell response and decline in PSA. Combined cryoablation with granulocyte macrophage colony-stimulating factor treatment was suggested to be an alternative approach for metastatic hormone refractory prostate cancer and could induce tumor-specific immunologic response. PMID- 19307998 TI - Combining an EGF-based cancer vaccine with chemotherapy in advanced nonsmall cell lung cancer. AB - An epidermal growth factor (EGF) vaccine was given before and after standard first line chemotherapy to patients with advanced nonsmall cell lung cancer (NSCLC), to investigate the immunologic and clinical results in a phase 1 study. Twenty patients diagnosed with advanced NSCLC were recruited. Two vaccinations were given before the first line of chemotherapy treatment, with subsequent monthly vaccination after concluding chemotherapy. The EGF vaccination dose was increased compared with previous trials; the primary end points were immunogenicity and safety. Anti-EGF antibody titers were more than 20 times higher than those previously obtained, without any increase in adverse events, serum EGF concentration decreased to undetectable levels in all patients. Ninety two percent of the evaluated patients (n=13) showed an immunodominant antibody response against the central region on the EGF molecule. High percentages of EGF/EGF receptor binding inhibition were observed, which significantly positively correlated with the increased antibody response against the EGF immunodominant region. Survival of the patients in this study correlates positively with antibody titers. This study has shown that combination of EGF vaccination at high dose, with chemotherapy is feasible and well tolerated higher anti-EGF antibody titers and reduction of serum EGF concentration seen; do not entail an increase in severe adverse events. The correlation of survival with antibody titers observed is being confirmed confirmation in a wider and randomized trial currently ongoing. PMID- 19308001 TI - [Digital versus analog mammography for breast cancer screening: critical methodological review of the literature and implications]. PMID- 19308002 TI - [Contrast enhanced MR angiography: evolving towards whole-body real time acquisitions]. AB - MRA includes all techniques used to depict vessels with MR. Gadolinium contrast injection combined with gradient echo sequences is the technique of choice for vascular imaging. Technical advances now allow faster acquisitions. The purpose of this article is to present two main advances with MRA: whole-body MRA and dynamic 3D MRA. Technical considerations, acquisition techniques, advantages and pitfalls based on our experience with a 1.5T MR unit will be discussed in order to promote their use in routine clinical practice. PMID- 19308003 TI - [CT imaging features and significance of gas in the pancreatic bed]. AB - The pancreas is an organ that normally does not contain gas. The purpose of this article is to reaffirm the value of CT to detect gas in abnormal locations and illustrate the different causes of gas collections in the pancreatic bed. Abscesses and infected pseudocysts are the most frequent causes of gas in the pancreatic bed followed by malignant and inflammatory fistulae. Iatrogenic etiologies should be considered along with the rare emphysematous pancreatitis associated with very poor prognosis. All of these entities shows multiple imaging findings, including the presence of gas in the pancreatic bed. PMID- 19308004 TI - [Osteonecrosis of the jaw and biphosphonates: imaging features]. AB - PURPOSE: 1) To review the pathophysiology of osteonecrosis of the jaw in patients receiving biphosphonates. 2) To review the imaging findings of osteonecrosis of the jaw and attempt to define pathognomonic imaging features. Materials and methods. Retrospective study of 15 patients with metastatic disease treated with biphosphonates. All available imaging studies including orthopantomograms, CT and bone scans were reviewed simultaneously by two radiologists (FO, DB). RESULTS: The most frequent imaging finding was osteolysis. Signs of biphosphonate impregnation were frequently observed: areas os osteosclerosis or heterogeneous demineralization due to abnormal bone remodeling. The outer cortex appeared duplicated in one case. Complications including fracture, sequestra, oroantral fistula and sinusitis may also occur. CONCLUSION: The imaging features of osteonecrosis remain fairly non-specific. Drug-related osteonecrosis of the jaw should nonetheless be suggested in the appropriate clinical setting in the presence of osteolysis associated with osteosclerosis. Imaging is helpful to assess the extent of the disease and detect complications for improved patient management. PMID- 19308005 TI - [CT exposure from pediatric MDCT: results from the 2007-2008 SFIPP/ISRN survey]. AB - PURPOSE: To evaluate current exposure levels from pediatric MDCT examinations in order to issue recommendations for the routine clinical practice that may be used for establishing future diagnostic reference levels (DRL). Materials and methods. A survey was conducted at hospital sites affiliated with the SFIPP (Societe Francophone d'Imagerie Pediatrique et Prenatale). Tube potential and volume computed tomography dose index (CTDI) vol. values were obtained for three age groups (1, 5 and 10 years) for typical scanning indications of eight anatomical regions. RESULTS: Twenty of 29 sites provided us with a copy of their protocols. All ages groups combined, 97% of protocols used a tension level < or =120 kVp. For age groups 1, 5 and 10 years respectively, the 75th percentiles of dose distributions were: 31, 39.5 and 49.5 mGy for the "head" protocol; 24, 22 and 24 mGy for "head and neck"; 11, 11, and 11 mGy for "paranasal sinuses"; 46, 71 and 87 mGy for "petrous bone"; 3, 3.5 and 5.5 mGy for "chest"; 2, 3 and 4 mGy for "low dose lung"; 4, 4.5 and 7 mGy for "abdomen-pelvis"; 7, 11 and 12 mGy for "bone". The inter-center discrepancy for doses were still high. CONCLUSION: This study provided an evaluation of current practices in reference centers for pediatric imaging and allowed readjustment of MDCT dose recommendations. It could help in the development of DRLS for pediatric MDCT. PMID- 19308006 TI - [US of the ulnal collateral ligament (UCL) at the first metacarpophalangeal (MCP) joint: a new dynamic maneuver to detect Stener lesions]. AB - Sprain of the first MCP joint is frequent. In patients with UCL tear, the adductor aponeurosis may become interposed between the distal site of ligament attachment and the ruptured ligament: this lesion, described by Stener, prevents ligamentous healing and requires surgical intervention. We present the results from a prospective study of 56 patients with first MCP sprain in order to determine the value of US by correlating with surgical findings. We propose a new dynamic maneuver during flexion-extension of the IP joint. This enables visualization of the adductor aponeurosis and its relationship to the UCL on US allowing detection of a Stener lesion when it exists. All patients with Stener lesion on US underwent surgery (32 operated patients): the sensitivity of US was 95.4% with a specificity of 80% for detection of Stener lesions. US, with the use of this specific dynamic maneuver is a reliable and reproducible tool for detecting Stener lesions. Review of video recordings allows visual comprehension of the examination for other health care providers. PMID- 19308007 TI - [Role of endovascular treatment of symptomatic splanchnic artery stenoses in HIV patients: report of three cases]. AB - Patients with HIV or AIDS frequently present with GI symptoms, sometimes due to early and diffuse atherosclerosis. We report 3 cases of HIV patients with abdominal pain due to severe splanchnic arterial stenosis. Only one patient presented typical clinical findings of mesenteric ischemic. Endovascular treatment was performed in all three cases. Good clinical outcome was immediate in 2 cases. In the third case, subsequent bowel resection was required due to irreversible ischemic injury in spite of local thrombolysis and endovascular revascularization in a patient presenting with acute severe mesenteric ischemia. In all three cases, vascular patency was demonstrated at follow-up. Mesenteric ischemia is a severe complication requiring early diagnosis in HIV patients, especially those with vascular risk factors, especially since endovascular treatment is a valid therapeutic option. PMID- 19308008 TI - [Medial foot pain in a marathon runner: a rare form of synostosis]. PMID- 19308009 TI - [Colonic wall hematoma complicating vasculitis]. PMID- 19308010 TI - [Intracranial epidermoid cyst secondary to mucocele surgery]. PMID- 19308011 TI - [Listeria rhombencephalitis: MR imaging features. A report of two cases]. PMID- 19308012 TI - [Answer to january e-quid: incidentally found scimitar syndrome]. PMID- 19308015 TI - What is the role of B-type natriuretic peptide in detecting heart failure in critically ill, mechanically ventilated patients? PMID- 19308016 TI - Fiberscopic-assisted placement of sheath of Fogarty embolectomy catheter as a guide to intubation through the ProSeal laryngeal mask airway. PMID- 19308017 TI - Cancer borealis stomatogastric nervous system dissection. AB - The stomatogastric ganglion (STG) is an excellent model for studying cellular and network interactions because it contains a relatively small number of cells (approximately 25 in C. borealis) which are well characterized. The cells in the STG exhibit a broad range of outputs and are responsible for the motor actions of the stomach. The stomach contains the gastric mill which breaks down food with three internal teeth, and the pylorus which filters the food before it reaches the midgut. The STG produces two rhythmic outputs to control the gastric mill and pylorus known as central pattern generators (CPGs). Each cell in the STG can participate in one or both of these rhythms. These CPGs allow for the study of neuromodulation, homeostasis, cellular and network variability, network development, and network recovery. The dissection of the stomatogastric nervous system (STNS) from the Jonah crab (Cancer borealis) is done in two parts; the gross and fine dissection. In the gross dissection the entire stomach is dissected from the crab. During the fine dissection the STNS is extracted from the stomach using a dissection microscope and micro-dissection tools (see figure 1). The STNS includes the STG, the oesophageal ganglion (OG), and the commissural ganglia (CoG) as well as the nerves that innervate the stomach muscles. Here, we show how to perform a complete dissection of the STNS in preparation for an electrophysiology experiment where the cells in the STG would be recorded from intracellularly and the peripheral nerves would be used for extracellular recordings. The proper technique for finding the desired nerves is shown as well as our technique of desheathing the ganglion to reveal the somata and neuropil. PMID- 19308019 TI - Effects of the Val158Met catechol-O-methyltransferase polymorphism on cortical structure in children and adolescents. PMID- 19308020 TI - The diacylglycerol kinase eta gene and bipolar disorder: a replication study in a Sardinian sample. PMID- 19308021 TI - Findings from bipolar disorder genome-wide association studies replicate in a Finnish bipolar family-cohort. PMID- 19308022 TI - Network analysis of positional candidate genes of schizophrenia highlights myelin related pathways. PMID- 19308024 TI - Regulation of cerebral blood flow in untreated mild-to-moderate hypertension. PMID- 19308025 TI - Is there a relationship between prenatal depression and preeclampsia? PMID- 19308026 TI - Fingerprinting the suspect: is there a smoking gun? PMID- 19308027 TI - Prenatal programming: maybe not so hopeless after all? PMID- 19308028 TI - Uric acid and the developmental origins of hypertension. PMID- 19308029 TI - Does the DASH diet improve clinical outcomes in hypertensive patients? PMID- 19308030 TI - Inflammatory activation and left ventricular mass in essential hypertension. AB - BACKGROUND: Inflammation is an independent risk factor for high blood pressure, and as a consequence inflammatory cytokines could be related with left ventricular hypertrophy (LVH). We sought to assess the association and predictive role of different cytokine levels with LVH in a group of patients with essential hypertension (HT). METHODS: We studied 251 asymptomatic hypertensive patients (142 with LVH and 109 without LVH), referred from 11 hospitals. A routine physical examination, laboratory analyses, and echo-Doppler study were performed. Plasma soluble tumor necrosis factor (TNF) receptors (sTNF-R1 and sTNF-R2), interleukin-6 (IL-6), and interleukin-1 receptor antagonist (IL-1ra) were centrally determined. RESULTS: Hypertensive patients with LVH had higher inflammatory cytokine levels than the group without hypertrophy (P < 0.001). Multivariate linear regression reported that sTNF-R1 (P < 0.01) was an independent predictor of left ventricular mass index (LVMI). All cytokines had significant area under the curves for detection of LVH, but sTNF-R1 has the highest area, 0.71 +/- 0.03 (P < 0.001). Finally, prevalence of LVH was increased in the group of patients with higher cytokine levels, and logistic regression analysis showed that sTNF-R1 (odds ratio = 2.59, 95% CI of 1.14-5.87) was an independent predictor of LVH. CONCLUSIONS: Cytokine levels were significantly correlated with LVMI in hypertensive patients. The sTNF-R1 was an independent predictor of LVMI. Plasma sTNF-R1 concentrations could be a predictive factor of LVH in patients with essential HT. PMID- 19308031 TI - Maternal periodontal disease and risk of preeclampsia: a case-control study. AB - BACKGROUND: We examined whether pregnant women with periodontal disease have an increased risk of preeclampsia, and we empirically evaluated the extent to which associations between periodontal disease and preeclampsia are dependent upon diagnostic criteria used to define periodontal disease operationally. METHODS: One hundred and fifty preeclampsia cases and one-fifty normotensive controls who delivered a singleton infant at term were enrolled. Periodontal examinations were performed within 48 h after delivery. Participants' periodontal health status was classified, a priori, into four categories according to the extent and severity of periodontal disease. Putative risk factors for periodontal disease and preeclampsia were ascertained during in-person postpartum interviews using a structured questionnaire and by medical record abstraction. Logistic regression was used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULTS: No clinically meaningful differences were observed between cases and controls with regard to periodontal parameters. After controlling for known confounders, severe clinical periodontal disease was not associated with an increased risk of preeclampsia (OR = 0.92, 95% CI: 0.26-3.28). In addition, there was no evidence of a linear increase in risk of preeclampsia with increasing severity of periodontal disease (P for trend = 0.65). When different diagnostic criteria previously used in other studies were used, the prevalence of periodontal disease varied substantially. However, the magnitude and direction of associations between periodontal disease and preeclampsia were largely similar regardless of the diagnostic criteria used to define periodontal disease. CONCLUSIONS: This study provides no convincing evidence that periodontal disease is associated with preeclampsia risk among Thai women. PMID- 19308033 TI - Severe hepatocellular injury after hematopoietic cell transplant: incidence, etiology and outcome. AB - Hepatic complications of transplant are a common cause of mortality. Although mild elevations of serum aminotransferase enzymes (aspartate and alanine (AST, ALT)) do not carry an adverse prognosis, this is not the case with severe hepatocellular injury. We reviewed 6225 consecutive recipients to determine the incidence and outcomes of severe hepatocellular injury (AST >1500 U/l) before day 100, which occurred in 88 patients. Causes were sinusoidal obstruction syndrome (SOS) (n = 46), hypoxic hepatitis (n = 33), varicella zoster virus (VZV) hepatitis (n = 4), drug-liver injury (n = 2) and unknown (n = 3). The incidence declined from 1.9% in the 1990s to 1.1% recently (owing to a fivefold decline in SOS and disappearance of VZV hepatitis). In hypoxic hepatitis, peak serum AST was 3545 U/l (range, 1380-25 246) within days of shock or prolonged hypoxemia; case fatality rate was 88%. In SOS, AST increases occurred 2-6 weeks after diagnosis; peak AST was 2252 U/l (range, 1437-8281); case fatality rate was 76%, with only serum bilirubin able to distinguish survivors (2.7 vs 11.3 mg/100 ml, P=0.0009). We conclude that circulatory insults (sinusoidal injury, hypotension and hypoxemia), and not infection, are the most common cause of severe hepatocellular injury, the frequency of which has declined because of a falling incidence of SOS and VZV hepatitis. PMID- 19308034 TI - Impact of PCR-based diagnosis of invasive pulmonary aspergillosis on clinical outcome. AB - The mortality rate of 60-90% in invasive pulmonary aspergillosis (IPA) is partly explained by diagnostic delay due to the limitation of current diagnostic tests. We assessed the influence of Aspergillus species (ASP) DNA detection by PCR from bronchoalveolar lavage (BAL) fluid, a new tool for diagnosing IPA, on the outcome of this disease in immune-compromised patients. The study population comprised 107 consecutive patients with hematological malignancies from a single medical center with IPA diagnosed between 1998 and 2005. Clinical variables and mortality rates were compared between two groups diagnosed according to traditional criteria without and with PCR-based ASP DNA detection in BAL fluid. The overall mortality rate during the study period was 38.3%. The addition of PCR to the diagnostic criteria shifted 31 patients from possible to probable IPA. Patients diagnosed with probable IPA according to traditional microbiological methods had significantly higher mortality rates compared to their counterparts who had in addition a PCR-based diagnosis (80 vs 35.6%, P=0.003). This study demonstrates that PCR-based ASP DNA detection for a diagnosis of IPA from BAL fluid has a significant effect on the outcome of patients with IPA, probably related to earlier diagnosis. PMID- 19308035 TI - Multipotent mesenchymal stromal cells for autoimmune diseases: teaching new dogs old tricks. AB - MSCs, otherwise known as multipotent mesenchymal stromal cells, are being examined for the treatment of autoimmune disease (AD) on the basis of their in vitro antiproliferative properties, efficacy in animal models, apparent low acute toxicity and the early positive anecdotal outcomes in human acute GVHD. Phase I/II clinical trials are underway in Crohn's disease and multiple sclerosis (MS) and are being planned for systemic lupus erythematosus (SLE), systemic sclerosis (SSc), systemic vasculitis and other AD. Open issues include patient selection, disease stage and activity, MSC source and expansion and long-term safety. Multidisciplinary groups are collaborating to ensure maximal use of available resources to establish the place, if any, of MSC in the treatment of AD. PMID- 19308036 TI - Factors predicting allogeneic PBSCs yield after G-CSF treatment in healthy donors. PMID- 19308037 TI - Ethical reasoning about patient eligibility in allogeneic BMT based on psychosocial criteria. AB - Chairpersons of the hospital ethics committees (HECs) and BMT clinicians were compared with regard to their willingness to proceed with allogeneic BMT given select psychosocial risk factors. A self-administered questionnaire was sent to 62 HEC chairpersons at hospitals with an accredited BMT program; the response rate was 37%. Items included background information, followed by six case vignettes from a 2006 national survey on which BMT physicians, nurses and social workers agreed not to proceed with allogeneic BMT on the basis of the following risk factors: suicidal ideation; use of addictive, illicit drugs; history of non compliance; absence of a caregiver; alcoholism; and mild dementia from early onset of Alzheimer's disease. Opinions regarding transplant differed in one case only, in a patient with mild dementia; 27% of HEC chairpersons recommended not proceeding with BMT, which was significantly lower than that of nurses (68%, P<0.001), physicians (63.5%, P<0.001) and social workers (51.9%, P=0.05). Qualitative data show patterns of informal reasoning, linking transplant decisions to patient's responsibility for their psychosocial risk factor(s), as well as to medical benefit and outcome. PMID- 19308038 TI - Pathological evidence of Wolman's disease following hematopoietic stem cell transplantation despite correction of lysosomal acid lipase activity. PMID- 19308039 TI - Disorders of glucose homeostasis in young adults treated with total body irradiation during childhood: a pilot study. AB - Impairment of glucose metabolism (in particular insulin resistance and type 2 diabetes mellitus) has been reported in patients who have undergone hematopoietic SCT (HSCT) during childhood, especially those treated with TBI. This pilot study was conducted to determine prevalence of and possible underlying mechanisms for impaired glucose homeostasis in young adults treated with HSCT and TBI and who were not previously known to have diabetes mellitus. A total of 10 subjects (6 males, 4 females) were evaluated. Mean ages were 13.0+/-1.0 years at the time of TBI and 24.0+/-1.1 years at the time of this study. Five subjects had laboratory evidence of insulin resistance using the homeostasis model assessment and the quantitative insulin sensitivity check index indices. Two of these subjects had impaired fasting glucose and four had decreased plasma insulin-like growth factor 1 levels. All 10 subjects had evidence of abdominal obesity. Insulin resistance is frequently observed in adult survivors of HSCT treated with TBI in childhood. Underlying mechanisms may include radiation-induced growth hormone deficiency and changes in body composition. PMID- 19308040 TI - Autologous stem cell transplants in Jehovah's Witnesses. PMID- 19308041 TI - Reduced intensity versus full myeloablative stem cell transplant for advanced CLL. AB - CLL remains incurable with the standard therapy. Allogeneic hematopoietic stem cell transplant may be curative. We examined 50 patients with advanced CLL who underwent allogeneic HCT at the University of Michigan between 1996 and 2006. Twenty-one patients received reduced-intensity conditioning (RIC) and twenty-nine patients received full-intensity conditioning (FIC) consisting of CY, etoposide and BCNU (n=20) or BU and CY (n=9). RIC recipients were older than FIC recipients (median age 54 vs 51, P=0.009). There were no statistically significant differences between groups in terms of the number of earlier therapies or patients with adverse cytogenetics. There were more unrelated donors in the RIC group 62% than in the FIC group 31% (P=0.030). Despite their older age and greater use of URD, the 5-year overall survival (OS) rate was 63% in the RIC group as compared with 18% in the FIC group (P=0.006). The primary cause of inferior survival in the FIC recipients was TRM, which was twice as high at day 100 for the FIC group 27% compared with the RIC group 14% (P=0.005). The relapse rate was 15% regardless with the majority of relapses occurring after day 100. These results suggest a favorable outcome for advanced CLL who undergo a RIC regimen compared with FIC. PMID- 19308042 TI - Risk factors for invasive aspergillosis and related mortality in recipients of allogeneic SCT from alternative donors: an analysis of 306 patients. AB - Invasive aspergillosis (IA) is a serious complication in patients undergoing allogeneic haematopoietic stem cell transplantation (HSCT), particularly from donors other than HLA-identical sibling. All 306 patients who underwent alternative donor HSCT between 01 January 1999 and 31 December 2006 were studied. Late IA was defined as occurring >or=40 days after HSCT. The median follow-up was 284 days (range, 1-2709). Donors were matched unrelated (n=185), mismatched related (n=69), mismatched unrelated (n=35) and unrelated cord blood (n=17). According to European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria, 2 patients already had IA at HSCT, 23 had early IA and 20 had late IA (IA incidence 15%). Eight patients had proven and 37 probable IA. Multivariate analyses showed that significant predictors of IA were delayed neutrophil engraftment, extensive chronic GVHD (cGVHD), secondary neutropenia and relapse after transplant. Early IA was associated with active malignancy at HSCT, CMV reactivation and delayed lymphocyte engraftment. Late IA was predicted by cGVHD, steroid therapy, secondary neutropenia and relapse after HSCT. IA-related mortality among IA patients was 67% and was influenced by use of anti-thymocyte globulin, steroids, higher levels of creatinine, and lower levels of IgA and platelets. The outcome of IA depends on the severity of immunodeficiency and the status of the underlying disease. PMID- 19308043 TI - Pegfilgrastim for PBSC mobilization and autologous haematopoietic SCT. AB - To date, G-CSF is the most favoured cytokine administered for PBSC mobilization because of its great efficacy and lack of serious toxicity. Recently, a pegylated filgrastim (pegfilgrastim) has been introduced. Attachment of the polyethylene glycol (PEG) moiety reduces renal excretion and masks proteolytic cleavage sites resulting in elevated G-CSF serum levels for up to 14 days after a single injection. As single-dose pegfilgrastim had similar effects in the prophylaxis of chemotherapy-induced neutropenia as the daily administration of the unconjugated drug, its capability for the mobilization of haematopoietic stem and progenitor cells has been assessed and presented to be at least equal to that of conventional G-CSF. Administration of pegfilgrastim following high-dose therapy and autologous blood SCT (BSCT) shortened the time to myeloid recovery as seen in conventional G-CSF. Plasma G-CSF levels were about 1 log higher with pegfilgrastim, but in the setting of autologous BSCT this did not translate into a faster haematopoietic recovery. Only few data exist on the biological effects of pegfilgrastim. Still, these data suggest that pegfilgrastim-stimulation results in different functional properties of haematopoietic stem and progenitor cells compared with G-CSF. PMID- 19308044 TI - NGAL decreases E-cadherin-mediated cell-cell adhesion and increases cell motility and invasion through Rac1 in colon carcinoma cells. AB - Expression of neutrophil gelatinase-associated lipocalin (NGAL)/lipocalin2, a recently recognized iron regulatory protein that binds to matrix metalloproteinase-9 (MMP9), is increased in a spectrum of cancers, including those of the colorectum. Using colon carcinoma cell lines stably transfected with NGAL or antisense NGAL, we showed that NGAL overexpression altered subcellular localization of E-cadherin and catenins, decreased E-cadherin-mediated cell-cell adhesion, enhanced cell-matrix attachment, and increased cell motility and in vitro invasion. Conversely, a decrease in NGAL enhanced more aggregated growth pattern and decreased in vitro invasion. We further showed that NGAL exerted these effects through the alteration of the subcellular localization of Rac1 in an extracellular matrix-dependent, but MMP9-independent, manner. Furthermore, we observed that the NGAL-overexpressing cells tolerated increased iron levels in the culture environment, whereas the NGAL-underexpressing cells showed significant cell death after prolonged incubation in high-iron condition. Thus, overexpressing NGAL in colon carcinomas is an important regulatory molecule that integrates extracellular environment cues, iron metabolism, and intracellular small GTPase signaling in cancer migration and invasion. NGAL may therefore be a new target for therapeutic intervention in colorectal carcinoma. PMID- 19308045 TI - Mechanisms of parenchymal injury and signaling pathways in ectatic ducts of chronic pancreatitis: implications for pancreatic carcinogenesis. AB - The pathobiology of chronic pancreatitis (CP) remains enigmatic despite remarkable progress made recently in uncovering key mechanisms involved in the initiation and progression of the disease. CP is increasingly thought of as a multifactorial disorder. Apoptosis plays a role in parenchymal destruction, the pathological hallmark of CP. The apoptotic mechanisms preferentially target the exocrine compartment, leaving endocrine islets relatively intact for a prolonged period. Exocrine cells shed their 'immunoprivileged' status, express death receptors, and are rendered susceptible to apoptosis induced by death ligands on infiltrating lymphocytes, and released locally by activated pancreatic stellate cells. Islet cells retain their 'immunoprivileged' status and activate anti apoptotic programs through NF-kappaB. Ductal changes, including distortion, dilatation, and pancreatic ductal hypertension in the setting of CP, induce genomic damage and increased cell turnover. In addition, signaling mechanisms that play a role in the development of embryonic pancreas are reinstated, thus, playing a role in repair, regeneration, and transformation. This, in turn, leads to acino-ductal metaplasia (ADM) and pancreatic intraepithelial neoplasia (PanIN). Some of these pathways are activated in pancreatic cancer. We attempt to integrate the current knowledge and major concepts in the pathogenesis of CP and to explain the mechanism of differential cell loss. We also discuss the possible implications of signaling pathway activation in pancreatic inflammation, relevant to the cellular transformation that leads to pancreatic neoplasia. PMID- 19308046 TI - Indoleamine 2,3-dioxygenase overexpression causes kynurenine-modification of proteins, fiber cell apoptosis and cataract formation in the mouse lens. AB - Indoleamine 2,3-dioxygenase (IDO) is the first enzyme in the kynurenine pathway. The kynurenines formed in this pathway chemically modify proteins and cause apoptosis in cells. Evidence suggests that kynurenines and their protein modifications are involved in cataract formation, but this has yet to be directly demonstrated. We generated transgenic (Tg) mouse lines that overexpress human IDO in the lens. Homozygous Tg (homTg) lenses had higher IDO immunoreactivity, approximately 4.5 times greater IDO mRNA, and approximately 8 times higher IDO activity compared to lenses from hemizygous Tg (hemTg) animals. The kynurenine content was threefold higher in homTg than in hemTg but was not detected in wild type (Wt) lenses. Kynurenine modifications were approximately 2.6 times greater in homTg than in hemTg or Wt. HomTg lenses had vacuoles in the epithelium and cortical fiber cells. Kynurenine modifications coincided with apoptosis in the secondary fiber cells of homTg lenses. Caspase-3 and caspase-9 activities were markedly higher in homTg than in hemTg and Wt. The glutathione content was approximately 36% lower in homTg compared to hemTg and Wt lenses. HomTg animals also developed bilateral cataracts within 3 months of birth. Together these data demonstrate that IDO-mediated production of kynurenines results in defects in fiber cell differentiation and their apoptosis and suggest that IDO activity is kept low in the lens to prevent deleterious effects by kynurenines. PMID- 19308047 TI - Increased expression of metastasis-related genes in hypoxic cells sorted from cervical and lymph nodal xenograft tumors. AB - Solid tumors contain regions of poor oxygenation that relate to the abnormal vascular network. Clinical investigations in cervical carcinoma have shown that positive lymph node status in patients with cervical carcinoma correlates with hypoxia. Earlier, in an orthotopic cervical cancer model, we had shown that exposure to acute hypoxia enhances lymph node metastasis. This study describes a technique for sorting hypoxic cells directly from the cervical xenograft model and reports the expression of 'metastasis-related' genes in hypoxic cells from xenografted cervix and lymph node tumors. Tumor cells were sorted on the basis of DsRed fluorescence and the sub-population of hypoxic cells was sorted on the basis of carbonic anhydrase-9 (CA-9) expression. Quantitative RT-PCR was conducted to measure changes in gene expression in the hypoxic cells sorted from primary cervix tumors and lymph node metastases. Immunohistochemistry was used to track changes in protein expression in sections of the same tumors. Metastasis related genes, CXCR4, uPAR, VEGFC, Hdm2, and OPN, were observed to be upregulated at gene and protein levels in the primary tumors and nodal metastasis from the orthotopic transplants. In particular, the hypoxic cells sorted from orthotopically transplanted cervix tumors and their lymph node metastases from mice exposed to cyclic (intermittent) hypoxia showed higher levels of expression of these genes. These results are consistent with the hypothesis that these genes may be involved in regulating lymph node metastasis in cervical cancers under hypoxic conditions and provide support to the concept cyclic hypoxia that plays an important role in this process. Our methodological study emphasizes the technique of cell sorting to identify hypoxic cells using CA-9, which may aid in improving prognostic capabilities and in designing rational therapeutic strategies by focusing on hypoxia-specific gene expression profiles of patients. The technique can be applied to identify other potential 'hypoxia-related' genes of interest for tumor growth and metastasis. PMID- 19308053 TI - Gender spotting and chimeric mice. PMID- 19308054 TI - Multiple campuses, one IACUC: how many AVs? Acting AVs. PMID- 19308055 TI - Multiple campuses, one IACUC: how many AVs? PMID- 19308056 TI - Multiple campuses, one IACUC: how many AVs? Delegate authority. PMID- 19308057 TI - Multiple campuses, one IACUC: how many AVs? A word from OLAW and USDA. PMID- 19308058 TI - Multiple campuses, one IACUC: how many AVs? Campus AVs plus an institutional veterinarian. PMID- 19308059 TI - Intrathoracic mass in a male Sprague-Dawley rat: thymoma. PMID- 19308061 TI - Novel food items as environmental enrichment for rodents and rabbits. AB - This column discusses the use of novel dietary supplements as environmental enrichment for rodents and rabbits. The purpose of enrichment is to encourage behaviors that are appropriate for a particular species and that satisfy an animal's physical and psychological needs. PMID- 19308062 TI - Efficacy of auditory enrichment in a prosimian primate (Otolemur garnettii). AB - Research suggests that auditory environmental enrichment might reduce abnormal behavior in certain primate species. The authors evaluated the behavioral effects of exposure to music in a prosimian primate (Garnett's bushbaby; Otolemur garnettii). They exposed bushbabies to a Mozart concerto for 15 min per day for 20 d (5 h exposure total), video-recorded them and subsequently analyzed the frequency of subjects' grooming and stereotypic behaviors. The authors compared the data with baseline behavioral data that had been recorded over a 20-d period before the experimental treatment. Neither stereotypy nor grooming behavior varied as a result of exposure to music. These results do not support the hypothesis that auditory enrichment in the form of exposure to music is an effective means of reducing stereotypic behavior in O. garnettii. PMID- 19308063 TI - Comparison of systemic Listeria monocytogenes infection in esophageally inoculated mice anesthetized with isoflurane or pentobarbital. AB - In previous attempts to produce a murine model of gastrointestinal listeriosis, the authors observed that pentobarbital anesthesia greatly increased disease severity in mice that were esophageally inoculated with the bacterium Listeria monocytogenes. In this study, they sought to evaluate the severity of systemic infection in inoculated mice that were anesthetized with isoflurane, an inhalational agent that is safer than pentobarbital and more commonly used for rodent anesthesia. Compared with pentobarbital anesthesia, isoflurane anesthesia resulted in infections of lesser severity, similar to those observed in unanesthetized mice. A pilot study in which mice were anesthetized with isoflurane for 5, 10 or 20 min suggested that this effect was not related to the duration of anesthesia. These results show that isoflurane anesthesia administered for 5 to 20 min does not potentiate the severity of listeriosis infection in mice esophageally inoculated with L. monocytogenes. PMID- 19308064 TI - The human-animal bond with laboratory animals. AB - In the author's experience, a bond--whether intentional or not--is often established between humans and animal research subjects. Behavioral theorists suggest that human-animal relationships can take several different forms. The author discusses several occasions in which she perceived one type of relationship with a research animal and was later surprised when the animal's behavior suggested that the bond was actually of a different nature. In the author's opinion, laboratory staff should be aware of the potential of bonding and should work to ensure that the desired human-animal relationships are developed. This may enhance the well-being of animals and staff and improve research efficacy. PMID- 19308065 TI - Data sharing in genomics--re-shaping scientific practice. AB - Funding bodies have recently introduced a requirement that data sharing must be a consideration of all funding applications in genomics. As with all new developments this condition has had an impact on scientific practice, particularly in the area of publishing and in the conduct of research. We discuss the challenges that must be addressed if the full benefits of data sharing, as envisaged by funders, are to be realized. PMID- 19308066 TI - Linking DNA methylation and histone modification: patterns and paradigms. AB - Both DNA methylation and histone modification are involved in establishing patterns of gene repression during development. Certain forms of histone methylation cause local formation of heterochromatin, which is readily reversible, whereas DNA methylation leads to stable long-term repression. It has recently become apparent that DNA methylation and histone modification pathways can be dependent on one another, and that this crosstalk can be mediated by biochemical interactions between SET domain histone methyltransferases and DNA methyltransferases. Relationships between DNA methylation and histone modification have implications for understanding normal development as well as somatic cell reprogramming and tumorigenesis. PMID- 19308067 TI - Metastasis: from dissemination to organ-specific colonization. AB - Metastasis to distant organs is an ominous feature of most malignant tumours but the natural history of this process varies in different cancers. The cellular origin, intrinsic properties of the tumour, tissue affinities and circulation patterns determine not only the sites of tumour spread, but also the temporal course and severity of metastasis to vital organs. Striking disparities in the natural progression of different cancers raise important questions about the evolution of metastatic traits, the genetic determinants of these properties and the mechanisms that lead to the selection of metastatic cells. PMID- 19308068 TI - The metastatic niche: adapting the foreign soil. AB - The 'seed and soil' hypothesis for metastasis sets forth the concept that a conducive microenvironment, or niche, is required for disseminating tumour cells to engraft distant sites. This Opinion presents emerging data that support this concept and outlines the potential mechanism and temporal sequence by which changes occur in tissues distant from the primary tumour. To enable improvements in the prognosis of advanced malignancy, early interventions that target both the disseminating seed and the metastatic soil are likely to be required. PMID- 19308069 TI - Parallel progression of primary tumours and metastases. AB - Systemic cancer progression is accounted for in two basic models. The prevailing archetype places the engine of cancer progression within the primary tumour before metastatic dissemination of fully malignant cells. The second posits parallel, independent progression of metastases arising from early disseminated tumour cells. This Perspective draws together data from disease courses, tumour growth rates, autopsy studies, clinical trials and molecular genetic analyses of primary and disseminated tumour cells in support of the parallel progression model. Consideration of this model urges review of current diagnostic and therapeutic routines. PMID- 19308070 TI - Children's acceptance of a peer who is overweight: relations among gender, age and blame for weight status. AB - OBJECTIVE: To examine the role of blame as a mediator of the relationships between perceiver age and gender and children's acceptance of an overweight peer. DESIGN: Cross-sectional study of children's perceptions of their overweight peers using structural equation modeling. PARTICIPANTS: Two hundred and ninety-one children between the ages of 3 and 11 years. MEASUREMENTS: Children viewed a videotape of a same-sex peer, dressed to appear overweight, interacting with an adult. After viewing the videotape, children responded to items assessing their perceptions of the child's social and emotional traits and how much the child was to blame for being overweight.Results:Confirmatory factor analysis indicated that the traits loaded on a single factor, acceptance. RESULTS: of analyses for our structural equation model indicated that as blame increased, acceptance of the peer decreased. The relationships between gender and blame and gender and acceptance were not significant. Children were categorized into three age groups (3-4, 5-8 and 9-11 years) to examine the influence of age. Children between 5 and 8 years of age were less likely to blame the model compared with younger and older children. CONCLUSION: Preschoolers reported the lowest acceptance, indicating a need for intervention for children in this age range. Furthermore, it will be important to conduct longitudinal studies to determine the influence of interventions as the child passes through different developmental stages. PMID- 19308071 TI - Meat consumption is associated with obesity and central obesity among US adults. AB - BACKGROUND: Meats are high in energy and fat content, and thus may be associated with higher risk of obesity. Many controversies remain regarding the association between meat consumption (MC) and obesity. OBJECTIVES: The aim of this study was to analyze the associations between MC and obesity assessed using body mass index (BMI > or = 30) and waist circumference (> or = 102 cm in men and > or = 88 cm in women) among US adults. METHODS: Nationally representative data collected in the 1999-2004 National Health and Nutrition Examination Survey (NHANES) were used. Linear and logistic regression analyses were conducted to test the associations between MC and adiposity measures controlling for potential confounders. RESULTS: Considerable differences existed in MC across sociodemographic groups among US adults. Those who consumed more meat had a much higher daily total energy intake, for example, those in the upper vs bottom quintiles consumed around 700 more kcal day(-1) (P<0.05). Regression models showed consistent positive associations between MC and BMI, waist circumference, obesity and central obesity, respectively. Using quintile 1 (low MC) as the reference, the association (odds ratio (OR) and 95% confidence interval (CI)) between total MC quintiles and obesity were 1.03 (0.88; 1.21; 2nd quintile), 1.17 (1.00; 1.38), 1.27 (1.08; 1.51) and 1.27 (1.08; 1.49; upper quintile), respectively; whereas that with central obesity was 1.13 (0.96-1.33), 1.31 (1.10; 1.54), 1.36 (1.17-1.60) and 1.33 (1.13; 1.55), respectively. CONCLUSIONS: These US national cross-sectional data show positive associations between MC and risk for obesity and central obesity. PMID- 19308072 TI - Injury of an aberrant vertebral artery during a routine corpectomy: a case report and literature review. AB - CASE REPORT: A case report of a 58-year-old man who sustained a laceration of his left vertebral artery during a routine corpectomy for cervical myelopathy is reported. OBJECTIVE: To report iatrogenic injury of a tortuous vertebral artery during anterior cervical spine surgery and discuss appropriate diagnosis and treatment options for this complication. SETTING: UMass Memorial Medical Center, Worcester, MA, USA. BACKGROUND DATA: Vertebral artery anomalies, although rare, are typically present with degenerative processes and great care must be taken to avoid damage during a corpectomy. Cross-sectional imaging coupled with intraoperative angiography is helpful for the urgent evaluation of the injury site and identification of the contralateral vertebral artery's status. METHODS: This is a single case of a patient sustaining a laceration of the left vertebral artery during surgery, which resulted in a lateral medullary stroke. RESULTS: After the left vertebral artery laceration, hemostasis was achieved. With the intent to better visualize and possibly embolize or stent the injury, an angiographic study was carried out. The angiogram revealed a laceration of the left vertebral artery within the vertebral foramina at vertebral body level C6, but intact distal flow. The patient underwent angiographic embolization and a subsequent magnetic resonance imaging (MRI) revealed a left lateral medullary stroke consistent with the lack of flow through the left vertebral artery from C6 to the basilar artery. CONCLUSION: If a tortuous vertebral artery is suspected, then meticulous review of preoperative cross-sectional imaging should be implemented along with angiographic examination. If anomalies are detected and the standard procedure cannot be safely carried out, then alterations, such as preoperative stent placement, need to be considered. PMID- 19308073 TI - Prospects for an influenza vaccine that induces cross-protective cytotoxic T lymphocytes. AB - Our approach to vaccination against influenza is unique. For no other pathogen do we construct and produce a new vaccine every year in the face of uncertainty about the strains that will be circulating when it is used. The huge global cooperative effort that underpins this process reflects our awareness of the need to control this major pathogen. Moreover, the threat of devastation by a pandemic due to a newly emerging viral subtype has triggered an intense effort to improve and accelerate the production of vaccines for use if a pandemic arises. However, type A influenza viruses responsible for seasonal epidemics and those with the potential to cause a pandemic share amino acid sequences that form the targets of cytotoxic T lymphocytes (CTL). CTL activated by currently circulating viruses, therefore, offer a possible means to limit the impact of infection with future variant seasonal strains and even new subtypes. This review examines how cross protective CTL can be exploited to improve influenza vaccination and issues that need to be considered when attempting to induce this type of immunity. We discuss the role of CTL responses in viral control and review the current knowledge relating to specificity and longevity of memory CD8(+) T cells, how vaccine antigen can be loaded into antigen-presenting cells to prime these responses and factors influencing the class of response induced. Application of these principles to the next generation of influenza vaccines should lead to much greater control of infection. PMID- 19308074 TI - Vaccines, global health and social equity. AB - Mortality rates of children less than 5 years old range from <10 per 1000 live births in industrialized countries to >100 in the world's poorest countries. The fact that in New York City infant mortality fell from approximately 140 deaths per 1000 live births in 1900 to <60 per 1000 by 1930 indicates what can be achieved through improvements in public health, such as the provision of clean piped water, sewage disposal and fresh refrigeration. For children living in impoverished conditions today, excess mortality is largely due to infectious diseases for which there are effective vaccines. Thus, certain specific vaccines can reduce mortality and morbidity, improve quality of life and contribute to economic development. However, because many vaccines and the means to deliver them are beyond the financial resources of countries with the highest childhood mortality rates, strategies have been devised to provide vaccines to the most needy populations. These strategies include initiatives by the Global Alliance for Vaccines and Immunization, the International Finance Facility for Immunization and the Advanced Market Commitment, which together with various governments, international agencies and charitable foundations are providing funds to make life-saving vaccines available to the world's most needy children. PMID- 19308075 TI - Lessons in science education. AB - Strong science education is an important part of any modern education. To ensure scientific progress, however, students need to aspire to academic careers. PMID- 19308076 TI - Hands-on inspiration for science. AB - To counter the decreasing interest in scientific studies, the PhysiScope at the University of Geneva conveys scientific excitement to teenagers by offering an entertaining and practical way to discover physics. PMID- 19308078 TI - Nanoparticle assembly: Anisotropy unnecessary. PMID- 19308079 TI - Material witness: Natural waterproofing. PMID- 19308080 TI - Complex oxides: A tale of two enemies. PMID- 19308081 TI - Nanomedicine: Veni, vidi, vici and then... vanished. PMID- 19308082 TI - Superconductivity: Commonalities in phase and mode. PMID- 19308083 TI - Electron and X-ray microscopy. PMID- 19308084 TI - Is science prepared for atomic-resolution electron microscopy? AB - The efforts of microscopists have given aberration-corrected transmission electron microscopy the power to reveal atomic structures with unprecedented precision. It is now up to materials scientists to use this power for extracting physical properties from microscopic atomic arrangements. PMID- 19308085 TI - Structure and bonding at the atomic scale by scanning transmission electron microscopy. AB - A new generation of electron microscopes is able to explore the microscopic properties of materials and devices as diverse as transistors, turbine blades and interfacial superconductors. All of these systems are made up of dissimilar materials that, where they join at the atomic scale, display very different behaviour from what might be expected of the bulk materials. Advances in electron optics have enabled the imaging and spectroscopy of these buried interface states and other nanostructures with atomic resolution. Here I review the capabilities, prospects and ultimate limits for the measurement of physical and electronic properties of nanoscale structures with these new microscopes. PMID- 19308086 TI - Electron tomography and holography in materials science. AB - The rapid development of electron tomography, in particular the introduction of novel tomographic imaging modes, has led to the visualization and analysis of three-dimensional structural and chemical information from materials at the nanometre level. In addition, the phase information revealed in electron holograms allows electrostatic and magnetic potentials to be mapped quantitatively with high spatial resolution and, when combined with tomography, in three dimensions. Here we present an overview of the techniques of electron tomography and electron holography and demonstrate their capabilities with the aid of case studies that span materials science and the interface between the physical sciences and the life sciences. PMID- 19308087 TI - Near-edge X-ray absorption fine-structure microscopy of organic and magnetic materials. AB - Many high-performance materials and novel devices consist of multiple components and are naturally or intentionally nano-structured for optimal properties and performance. To understand their structure-property relationships fully, quantitative compositional analysis at length scales below 100 nm is required, a need that is often uniquely addressed using soft X-ray microscopy. Similarly, the interaction of X-rays with magnetic materials provides unique element-specific contrast that allows the determination of magnetic properties in multi-element antiferromagnetic and ferromagnetic materials. Pump-probe-type experiments can even investigate magnetic domain dynamics. Here we review and exemplify the ability of soft X-ray microscopy to provide information that is otherwise inaccessible, and discuss a perspective on future developments. PMID- 19308088 TI - Coherent X-ray diffraction imaging of strain at the nanoscale. AB - The understanding and management of strain is of fundamental importance in the design and implementation of materials. The strain properties of nanocrystalline materials are different from those of the bulk because of the strong influence of their surfaces and interfaces, which can be used to augment their function and introduce desirable characteristics. Here we explain how new X-ray diffraction techniques, which take advantage of the latest synchrotron radiation sources, can be used to obtain quantitative three-dimensional images of strain. These methods will lead, in the near future, to new knowledge of how nanomaterials behave within active devices and on unprecedented timescales. PMID- 19308089 TI - X-ray imaging beyond the limits. AB - The intense, brief pulses of X-rays from upcoming free-electron lasers will greatly extend X-ray microscopy to the femtosecond time domain and to interatomic length scales. From recent experiments and simulations one can envisage imaging macromolecules with X-rays without the need for crystallization. PMID- 19308090 TI - Feeder-free maintenance of hESCs in mesenchymal stem cell-conditioned media: distinct requirements for TGF-beta and IGF-II. AB - A paracrine regulation was recently proposed in human embryonic stem cells (hESCs) grown in mouse embryonic fibroblast (MEF)-conditioned media (MEF-CM), where hESCs spontaneously differentiate into autologous fibroblast-like cells to maintain culture homeostasis by producing TGF-beta and insulin-like growth factor II (IGF-II) in response to basic fibroblast growth factor (bFGF). Although the importance of TGF-beta family members in the maintenance of pluripotency of hESCs is widely established, very little is known about the role of IGF-II. In order to ease hESC culture conditions and to reduce xenogenic components, we sought (i) to determine whether hESCs can be maintained stable and pluripotent using CM from human foreskin fibroblasts (HFFs) and human mesenchymal stem cells (hMSCs) rather than MEF-CM, and (ii) to analyze whether the cooperation of bFGF with TGF-beta and IGF-II to maintain hESCs in MEF-CM may be extrapolated to hESCs maintained in allogeneic mesenchymal stem cell (MSC)-CM and HFF-CM. We found that MSCs and HFFs express all FGF receptors (FGFR1-4) and specifically produce TGF-beta in response to bFGF. However, HFFs but not MSCs secrete IGF-II. Despite the absence of IGF-II in MSC-CM, hESC pluripotency and culture homeostasis were successfully maintained in MSC-CM for over 37 passages. Human ESCs derived on MSCs and hESCs maintained in MSC-CM retained hESC morphology, euploidy, expression of surface markers and transcription factors linked to pluripotency and displayed in vitro and in vivo multilineage developmental potential, suggesting that IGF-II may be dispensable for hESC pluripotency. In fact, IGF-II blocking had no effect on the homeostasis of hESC cultures maintained either on HFF-CM or on MSC-CM. These data indicate that hESCs are successfully maintained feeder-free with IGF-II-lacking MSC-CM, and that the previously proposed paracrine mechanism by which bFGF cooperates with TGF-beta and IGF-II in the maintenance of hESCs in MEF-CM may not be fully extrapolated to hESCs maintained in CM from human MSCs. PMID- 19308091 TI - BMP-6 inhibits microRNA-21 expression in breast cancer through repressing deltaEF1 and AP-1. AB - MicroRNAs (miRNAs), which are small noncoding RNA molecules, play important roles in the post-transcriptional regulation process. The microRNA-21 gene (miR-21) has been reported to be highly expressed in various solid tumors, including breast cancer. Bone morphogenetic protein-6 (BMP-6) has been identified as an inhibitor of breast cancer epithelial-mesenchymal transition (EMT) through rescuing E cadherin expression. We initiated experiments to identify the relationships between miR-21 and BMP-6 in breast cancer progression. Real-time PCR analysis showed that miR-21 expression was very high in MDA-MB-231 cells that expressed little BMP-6. A reverse correlation between BMP-6 and miR-21 was also determined in breast cancer tissue samples. Moreover, BMP-6 inhibited miR-21 transcription in MDA-MB-231 cells. In order to investigate how BMP-6 inhibited the miR-21 promoter (miPPR-21), we constructed a series of miPPR-21 reporters. Luciferase assay results indicated that BMP-6 inhibited miPPR-21 activity through the E2-box and AP-1-binding sites. We also demonstrated that both deltaEF1 and TPA induced miR-21 expression. Using site-directed mutation and CHIP assay, we found that deltaEF1 induced miPPR-21 activity by binding to the E2-box on miPPR-21. Moreover, TPA triggered miPPR-21 activity through the AP-1 binding sites. BMP-6 treatment significantly reduced the binding of these factors to miPPR-21 by decreasing the expression of deltaEF1 and c-Fos/c-Jun. We also demonstrated that BMP-6-induced downregulation of miR-21 modified the activity of PDCD4 3'UTR and inhibited MDA-MB-231 cell invasion. deltaEF1 overexpression and TPA induction blocked this inhibitory effect of BMP-6. In conclusion, BMP-6-induced inhibition of miR-21 suggests that BMP-6 may function as an anti-metastasis factor by a mechanism involving transcriptional repression of miR-21 in breast cancer. PMID- 19308093 TI - A landmark systems analysis of prion disease of the brain. PMID- 19308092 TI - A systems approach to prion disease. AB - Prions cause transmissible neurodegenerative diseases and replicate by conformational conversion of normal benign forms of prion protein (PrP(C)) to disease-causing PrP(Sc) isoforms. A systems approach to disease postulates that disease arises from perturbation of biological networks in the relevant organ. We tracked global gene expression in the brains of eight distinct mouse strain-prion strain combinations throughout the progression of the disease to capture the effects of prion strain, host genetics, and PrP concentration on disease incubation time. Subtractive analyses exploiting various aspects of prion biology and infection identified a core of 333 differentially expressed genes (DEGs) that appeared central to prion disease. DEGs were mapped into functional pathways and networks reflecting defined neuropathological events and PrP(Sc) replication and accumulation, enabling the identification of novel modules and modules that may be involved in genetic effects on incubation time and in prion strain specificity. Our systems analysis provides a comprehensive basis for developing models for prion replication and disease, and suggests some possible therapeutic approaches. PMID- 19308095 TI - Getting started. PMID- 19308096 TI - Journey to a journal: a vision to create a forum for Canadian health services and policy research--tracing the origins of Healthcare Policy. PMID- 19308098 TI - Baneful legacy: medicare and Mr. Trudeau: the Constitution created by the Trudeau government is now threatening Canada's medicare system. What can be done to defend it? PMID- 19308099 TI - Courting trouble: the Supreme Court's embrace of private health insurance: use and misuse of social science evidence by the Supreme Court--how should Canadian governments respond? PMID- 19308101 TI - Reflections on a conversation with Brian Postl: can healthcare research make a difference to policy and practice? Interview by Anton Hart. PMID- 19308100 TI - The Chaoulli judgment or how to sell off a public right. PMID- 19308102 TI - Are there socio-economic differences in caesarean section rates in Canada? AB - Caesarean section rates have risen in recent years, sparking renewed debate about the circumstances under which such deliveries are being, and should be, performed. Some commentators suggest that increasing rates may, in part, be explained by women in higher-income brackets requesting elective caesareans (the so-called "too posh to push" hypothesis). After adjusting for maternal age, Canadian data do not support this theory. In fact, age-adjusted caesarean section rates were significantly lower in Canada's highest-income neighbourhoods than in the lowest-income areas in 2002-03. PMID- 19308103 TI - Using research to inform healthcare managers' and policy makers' questions: from summative to interpretive synthesis. AB - This paper highlights the importance of research synthesis for healthcare managers' and policy makers' questions and the difficulty of generalizing from the methods used to answer clinicians' questions. Social science research has a central role in such syntheses because of the context-dependent nature of managers' and policy makers' questions, which generally encompass a far broader spectrum than the circumscribed "what works?" questions of clinically oriented reviews. A major challenge is in moving from purely researcher-driven processes, which summarize research, to co-production processes, which allow managers and policy makers to join with researchers in interpreting implications for the healthcare system. Additional challenges lie in clearly defining the function, role and objective of the synthesis; handling flexibility around finalizing the question; harnessing a manageable scope of literature to review; adopting rules to select the final sample of research; creating useful messages; and developing a format that is responsive to the needs and preferences of the audience. One inevitable conclusion is that research synthesis for managers and policy makers will, compared to that for clinicians, leave much discretion in the hands of the synthesiser(s). This raises the interesting issue of how to engender, in the absence of "methodological checklists," trust and credibility in both the people doing the synthesis and the processes they use. PMID- 19308104 TI - When health services researchers and policy makers interact: tales from the tectonic plates. AB - There has been a strong push over the last decade for health services researchers to become "relevant," to work with policy makers to translate evidence into action. What has been learned from this interaction? The pooled experiences of health services researchers across the country, including those at the Manitoba Centre for Health Policy (MCHP), suggest five key lessons. First, policy makers pay more attention to research findings if they have invested their own funds and time. Second, researchers must make major investments in building relationships with policy makers, because there are inevitable tensions between what the two parties need and do. Third, researchers must be able to figure out and communicate the real meaning of their results. Fourth, health services researchers need a "back-pocket" mindset, as they cannot count on immediate uptake of results; because the issues never go away, evidence, if known and easily retrievable, is likely to have an eventual impact. Finally, getting evidence into the policy process does not come cheaply or easily, but it can be done. The overriding lesson learned by health services researchers is the importance of relationship-building, whether in formalizing contractual relationships, building and maintaining personal trust, having a communications strategy or increasing the involvement of users in the research process. PMID- 19308105 TI - Drug expenditure trends in the Canadian provinces: magnitude and causes from 1998 to 2004. AB - This analysis uses a consistent pan-Canadian dataset--Canadian CompuScript from IMS Health, Canada--to quantify trends in per capita drug expenditures within each Canadian province over the period of 1998 to 2004. The impacts of changes in six potential determinants of drug expenditure are calculated for every province. Each of the six detailed cost drivers falls into one of three broad categories: volume effects, price effects and therapeutic choices. Despite wide variation in expenditure levels, the rate and causes of provincial expenditure trends over time were roughly comparable. From 1998 to 2004, per capita expenditures on oral solid prescription drugs grew at a rate of over 10% per year in most provinces- several times faster than economic growth over the same period. This rapid expenditure growth has largely been due to increased utilization of medicines and a trend towards prescribing higher-cost drugs over time. Price changes had little impact on drug spending in all provinces. PMID- 19308106 TI - Public drug plan coverage for children across Canada: a portrait of too many colours. AB - BACKGROUND: As debate continues regarding pharmacare in Canada, little discussion has addressed appropriate drug plan coverage for vulnerable populations, such as children. The primary objective of this study was to determine the extent of medication coverage for children in publicly administered programs in each province across Canada. METHODS: Data were collected on provincial, territorial and federal government drug plans, and 2003 formulary updates were obtained. A simulation model was constructed to demonstrate costs to a low-income family with an asthmatic child in each province. Programs were compared descriptively. The extent of interprovincial variation in 2003 formulary approvals was summarized statistically. RESULTS: There was 39% variation between provinces with respect to 2003 formulary approvals (chi-square p < 0.0001) and 48% variation for 2003 paediatric-labelled products (chi-square p < 0.0001). Across Canada, only 8% of 2003 formulary approvals were indicated primarily for paediatric conditions. In the simulation model, costs were less than or equal to 3% of household income in provinces with plans for low-income families, catastrophic costs (Ontario) or for the population. Families who failed to qualify for low income plans or who resided in New Brunswick or Newfoundland faced costs up to 7% of household income. INTERPRETATION: With regard to pharmaceutical benefits for children, provincial drug programs vary considerably in terms of whom they cover, what drugs are covered and how much subscribers must pay out of pocket. Unlike seniors and social assistance recipients, the provinces do not agree on the importance of providing comprehensive coverage for all children. For many Canadian children, significant financial barriers exist to medication access. PMID- 19308107 TI - Joining the conversation: newspaper journalists' views on working with researchers. AB - For health researchers who seek more research use in policy making to improve health and healthcare, working with the news media may represent an opportunity, given the media's pivotal role in public policy agenda-setting. Much literature on science and health journalism assumes a normative stance, focusing on improving the accuracy of news coverage. In this study, we investigated journalists' perspectives and experiences. We were particularly interested in learning how health researchers could work constructively with journalists as a means to increase research use in policy making. Qualitative methods were used to conduct and analyze interviews with experienced newspaper journalists across Canada, with children's mental health as a content example. In response, study participants emphasized journalistic processes more than the content of news coverage, whether children's mental health or other topics. Instead, they focused on what they thought researchers needed to know about journalists' roles, practices and views on working with researchers. Newspaper journalists balance business and social responsibilities according to their respective roles as editors, columnists and reporters. In practice, journalists must ensure newsworthiness, relevance to readers and access to sources in a context of daily deadlines. As generalists, journalists rely on researchers to be expert interpreters, although they find many researchers unavailable or unable to communicate with public audiences. While journalists are skeptical about such common organizational communications tools as news releases, they welcome the uncommon contributions of those researchers who cultivate relationships and invest time to synthesize and communicate research evidence on an ongoing basis. Some appealed for more researchers to join them in participating in public conversations. We conclude that there are opportunities for policy-oriented health researchers to work constructively with newspaper journalists--by appreciating journalists' perspectives and by taking seriously some of their suggestions for engaging in public conversations--and that such engagement can be a means to increase the use of research evidence in policy making and thereby improve health and healthcare. PMID- 19308108 TI - Coming of age and taking stock: the state of academic health policy research centres in Canada. AB - This descriptive study takes stock of the nation's health services and health policy research capacity by profiling the organizational models, operational challenges and success strategies utilized by Canadian academic health policy research centres. While each such centre is unique, the results point to some common themes, including symbiotic relationships between centres and their ministries of health, pervasive infrastructure funding challenges and the importance of having a supportive academic home. PMID- 19308109 TI - Population dynamics of plant nematodes in cultivated soil: effects of combinations of cropping systems and nematicides. AB - The population density of Meloidogyne incognita was significantly reduced in land that was fallowed or cropped to crotalaria, marigold, bermudagrass, or bahiagrass. The rate of population decline caused by different cropping systems was influenced by initial population densities. Crotalaria, marigold, and bare fallow were about equally effective in reducing the density of M. incognita below dctectable lcvels, usually requiring 1-3 yr. Bahiagrass and bcrmudagrass required 4-5 yr or longer to reduce M. incognita below a detectable level. A high population density of Trichodorus christiei developed in land cropped 5 yr to bermudagrass, bahiagrass, okra, and marigold. Population densities of Pratylenchus brachyurus and Xiphinema americanum increased in land cropped to crotalaria or bermudagrass. Belonolabnus Iongicaudatus was detected only in land cropped to bermudagrass, The effectiveness of nematicides in reducing M. incognita infection was rclatcd to nematode population density resulting from 5 yr of different cropping systems. Treatment with aldicarb reduced M. incognita below detectable levels following all cropping systems; treatment with ethoprop following all cropping systems except okra, treatment wflh ethylene dibromide following bahiagrass or fallow; and treatment with DBCP only after 5 yr of fallow. Tomato transplant growth was affected .by both cropping systems and nematicide treatment. Transplants grown after crotalaria and bahiagrass were significantly larger than those grown after other crops. Also, treatment with aldicarb and ethoprop significantly increased transplant size. PMID- 19308111 TI - A Description of Males of Hoplolaimus columbus. AB - The male of the Columbia lance nematode, Hoplolaimus columbus, is described and illustrated from a harvested soybean field in Holly Hill, South Carolina. It is morphologically similar to the female, except for reproductive structures. PMID- 19308110 TI - A Pest Management Approach to the Control of Pratylenchus thornei on Wheat in Mexico. AB - The lesion nematode, Pratylenchus thornei, was clearly demonstrated as a parasite of wheat. It reduced plant stands and stunted plants in the field under the environmental conditions found in Sonora, Mexico. Other soil organisms also may have contributed to the problem. The nematode is widely distributed throughout the wheat-growing region, and may be a problem each growing season. Nematicides controlled the nematode and increased yields, but they were not economical. No resistance was found in existing commercial wheat cultivars. A pest management approach using variety selection, nitrogen fertilizer, planting in cool soil (15 C) and a crop rotation avoiding wheat after wheat was the most practical solution to this problem on a commercial scale. PMID- 19308112 TI - Nematode Community Structure of Forest Woodlots: III. Ordinations of Taxonomic Groups and Biomass. AB - Nematode communities of 18 Indiana mixed hardwood stands were comprised of an average of 23% tylenchid species, 31% dorylaimid species, and 46% species of six other orders. Based on total numbers of individuals present the stands averaged 42% tylenchids, 20% dorylaimids, and 38% individuals of other orders. Ordination of the sites using data only for tylenchid species gave an even distribution of sites, indicating little effect of site disturbances on tylenchid populations. By contrast an ordination using data for dorylaimid species showed a high degree ofd issimilarity between reference sites indicating that disturbances at some sites had drastically affected the dorylaimid fauna. An ordination utilizing biomass of all species present was very similar to the ordination based on data for dorylaimid species only. PMID- 19308113 TI - Mass Culturing of Ditylenchus dipsaci to Yield Large Quantities of Inoculum. AB - Methods are described for rearing large quantities of Ditylenchus dipsaci on alfalfa tissues. Nematodes and alfalfa seed were disinfected and nematodes were reared in quantities sufficient to provide a continuous supply of inoculum for our alfalfa-breeding program. Nematodes reproduced best in darkness at 20-25 C. Cultures reached maximum numbers in 3-6 wk. PMID- 19308115 TI - Comparative stereoscan electron micrographs of nematode heads. PMID- 19308114 TI - Pristionchus lheritieri as a carrier of Rhizobium japonicum. PMID- 19308116 TI - Specificity of retention and transmission of viruses by nematodes. PMID- 19308117 TI - Nature and inheritance of nematode resistance in cereals. AB - Resistance to a number of nematodes is present in varieties of temperate and tropical cereals. The occurrence, nature, and inheritance of varietal resistance in cereals is reviewed. Evaluation of the practical significance of nematode resistance in a particular host-nematode combination is discussed in relation to host efficiency, host sensitivity, genetic control of resistance, and presence of virulence in the nematode population. PMID- 19308118 TI - Biochemical mechanisms of plant resistance to nematodes: a review. PMID- 19308119 TI - Integrated approach to the control of the golden nematode, heterodera rostochiensis. AB - Under high population densities of Heterodera rostochiensis, control is impossible by one method alone, whether by chemical means, crop rotation, or the use of resistant potato varieties. Integrated control using thiazone (DMTT) in autumn, resistant potato hybrid 61-8/1, or Solanum andigenum cultivars 'Antinema' and 'Specula', followed by cereal and clover reduced the population below the detection level in the upper 18-cm on small plots on sandy loam soil. PMID- 19308120 TI - Seasonal population dynamics of selected plant-parasitic nematodes on four monocultured crops. AB - Seasonal fluctuations in field populations of Meloidogyne incognita, Pratylenchus zeae, P. brachyurus, Criconemoides ornatus, Trichodorus christiei, and Helicotylenchus dihystera on monocultured corn, cotton, peanut, and soybean were determined monthly for 4 yr. Population densities of M. incognita were greater in corn and cotton plots than in peanut and soybean plots from July until January. Those of Pratylenchus spp. were greater on corn and soybean than on cotton and peanut during all months except May and June. C. ornatus populations were greater on corn and peanut than on cotton and soybean during all months. C. ornatus on corn and peanut was more numerous in July than in other months. There was no significant increase in populations of T. christiei, except on corn in June. H. dihystera was greater in cotton and soybean plots than in corn and peanut plots from August through December. PMID- 19308121 TI - Dynamics of Concomitant Field Populations of Hoplolaimus columbus and Meloidogyne incognita. AB - From the fall of 1968 through the summer of 1973, a Georgia cotton field with a lengthy history of the Cotton Stunt Disease Complex was sampled for the presence of plant parasitic nematodes. Although Meloidogyne incognita was recovered on all sampling dates, concomitant populations of Hoplolaimus columbus were not recovered until the spring of 1970. During the succeeding four growing seasons, the population density and horizontal distribution of H. columbus increased, and H. columbus replaced M. incognita as the predominant phytopathogenie species. A second Georgia cotton field containing concomitant populations of H. columbus and M. incognita was observed from the fall of 1971 through the summer of 1973. In this case the horizontal distribution of both species remained relatively constant and the population density of H. columbus increased steadily. In both locations, the presence of either H. columbus or M. incognita significantly inhibited the presence of the concomitant species. In general, however, the initial spring or final fall population densities of H. columbus or M. incognita had no significant influence on the population density of the concomitant species, The data are also discussed in relation to the biological significance of H. columbus in the southeastern coastal plain. PMID- 19308122 TI - Biochemical Changes in Root Exudate and Xylem Sap of Tomato Plants Infected with Meloidogyne incognita. AB - Under two monoxenic culture techniques of growing plants (filter paper and silica sand cultures), sugar in root exudate from Meloidogyne incognita-infected tomato increased 133 to 836% over controls. In contrast, amino acids were moderately reduced 52 to 56%. Chromatographic analysis showed that galled root exudate contained three sugars, twelve amino acids, and three organic acids, whereas healthy root exudate contained four sugars, fifteen amino acids, and four organic acids. Polysaccharide was responsible for the large increase of sugars in galled root exudates. The concn and the absolute amount of total sugars in the infected plant xylem sap were greater than in healthy plant xylem sap up to 6 wk after inoculation, whereas amino acids were moderately lower than in controls throughout the test period. Chromatographic analysis showed that xylem sap from both healthy and infected plants at 4 wk after inoculation contained four sugars and five organic acids. We identified 18 and 17 amino acids in the healthy and infected plant xylem sap, respectively. The concn of sugar increased as the nematode inoculum increased at 2, 4 and 6 wk after inoculation. The amino acids in all samples from the infected plant moderately decreased with an increase of nematode inoculum. We suggest that changes in total sugars and amino acids, of infected plant xylem sap and root exudate are a probable mechanism by which tomato plants are predisposed to Fusarium wilt. PMID- 19308123 TI - Seasonal fluctuations in the spatial distribution of nematode populations in a california vineyard. AB - Distribution of Xiphinema americanum and four Meloidogyne spp, was studied in a vineyard over a 13-mo. period. The X. arnericanum population was concd in the upper 60-cm of undisturbed soil in the vine row, whereas the Meloidogyne species were distributed both in and between rows and to greater depths, similar to the distribution of the root system. Samples for assessment of X. americanum densities had least variation when taken in the vine row from the upper 60-cm of soil. Sampling error is reduced in Meloidogyne populations by sampling within 40 cm of the vine both within and/or between rows. PMID- 19308124 TI - Discocriconemella inaratus n. sp. and Criconemoides inusitatus n. sp. (Nematoda) from Iowa. AB - Discocriconemella inaratus n. sp. from Iowa prairies is characterized by a single offset disk-shaped head annule which is often discontinuous, a sigmoid vagina, a stylet length of 51-61 mum, and 77-100 smooth body annules. Criconemoides inusitatus n. sp. from Iowa woodlands is characterized by two offset head annules, a "closed" vulva, a straight vagina, no overlapping anterior vulva lip, a stylet length of 42-50 mum, and 71-86 smooth body annules. PMID- 19308125 TI - Interrelationship of Aphelenchoides fragariae and Xanthomonas begoniae on Rieger begonia. PMID- 19308126 TI - A Rapid Method for Separating Larvae from Mixed Population of Panagrellus redivivus. PMID- 19308127 TI - Pathogenicity of Pratylenchus coffeae, Scutellonema bradys, Meloidogyne incognita, and Rotylenchus reniformis on Dioscorea rotundata. AB - Low populations (200 specimens per plant) of Pratylenchus cofl'eae, Scutellonema bradys, Meloidogyne incognita, and Rotylenchulus reniformis stimulated the development of tops, roots, and tubers of Dioscorea rotundata "Guinea" yam. We demonstrated experimentally that P. coffeae was responsible for the deterioration in quality of the yam tuber in Puerto Rico, a condition known as a dry rot of yam. Initial populations of 600 P. coffeae, S. bradys, or M. incognita, and populations of 1,000 P. coffeae or S. bradys per plant were high enough to induce dry rot of the yam tubers. P. coffeae and S. bradys were pathogenic to yam cultivar Guinea, but M. incognita and R. reniformis did not cause necrosis or cracking of the tuber cortex in our experiments. PMID- 19308128 TI - Effect of Meloidogyne incognita on Reproduction of Pratylenchus penetrans in Red Clover and Alfalfa. AB - Roots of seedlings of red clover and alfalfa growing on 10(1) Hoagland and Arnon solution agar were inoculated with various combinations of Meloidogyne incognita and Pratylenchus penetrans. Egg-laying by P. penetrans decreased as the number of nematodes, the ratio of entrant M. incognita to entrant P. penetrans, and the priority of invasion of roots by M. incognita increased. Embryogeny and hatching of eggs of P. penetrans, and development of larvae of M. incognita, were not affected. In red clover, the greatest red uction occurred when there were 65 entrant nematodes, the ratio of M. incognita:P. penetrans was 4:1 and M. incognita was inoculated four days prior to P. penetrans. In alfalfa, the less favorable host for both nematodes, the greatest reduction occurred when there were 45 entrant nematodes, the ratio of M. incognita:P. penetrans was 2:1, and M. incognita was inoculated 4 days prior to P. penetrans. PMID- 19308129 TI - Free Amino Acids in Roots of Infected Cotton Seedlings Resistant and Susceptible to Meloidogyne incognita. AB - Quantities of free amino acids in segments of cotton roots resistant and susceptible to Meloidogyne incognita were compared. Following infection, the root knot susceptible cultivar, M8, had greater percentage increases of certain individual free amino acids than the resistant cultivar, Clevewilt, but the sum total of free amino acids was greatest in the resistant cultivar. More free amino acids were present in infected than in noninfected plants of both cultivars. The overall concn of glycine declined over the I 0-day period following inoculation. The concns of the aromatic amino acids, tyrosine and phenylalanine, varied as functions of infection, cultivar, and time of harvest. Proline in susceptible M8 increased nearly 2000-fold 10 days after infection, when considerable thickening of syncytial walls is occurring. PMID- 19308130 TI - Revision of the genus paratylenchus micoletzky, 1922, and descriptions of new species. Part I of 3 parts. AB - A revision of part of the genus Paratylenchus is reported covering those species with stylets averaging < 22 mum. Thirteen new species are described and further observations are given on the morphology and relationships of nine other species. Paratylenchus gahriciis transferred to the genus Hemicriconemoides. This article is the first of three parts: the second will treat Paratylenchus spp. with stylets averaging 24-40 mum (and include a key to the species described in parts 1 and2), the third will include species with stylets >40mum, respectively. PMID- 19308131 TI - Oogenesis and the chromosomes of twelve bisexual species of heterodera (nematoda: heteroderidae). AB - Twelve bisexual species of Heterodera reproduced by amphimixis and had the same number of n=9 (2n=18) chromosomes in maturing oocytes. H. schachtii had slightly larger chromosomes than all other species. Only sperm nuclei with n=9 chromosomes were observed inside maturing oocytes and no specialized sex chromosomes were detected in any case. A "supernumerary" chromosome was observed occasionally in oocytes of H. schachtii and H. weissi and was transmitted regularly to one-half of the progeny of the nematodes that possessed it. Cytological characteristics were not very instructive in differentiating amphimictic tleterodera species. Such karyotypic uniformity indicates cytogenetic stability of the genus and close interrelationship among its members. PMID- 19308132 TI - Fine Structure of Cephalic Sense Organs in Heterodera glycines Males. AB - Cephalic sense organs of Heterodera glycines males were examined in detail by electron microscopy. Each amphid basically consists of an amphidial gland, a nerve bundle, and an amphidial duct. The amphidial gland consists of a microvillous region, and laterally is closely associated with a large secretory cell. The nerve bundle penetrates the microvillous region, and further anteriorly individual nerve processes (dendrites) separate from one another, thus forming a sensilla pouch which is enveloped by the microvillous region of the gland. Anterior to the pouch, the cilia-like dendrites converge as they enter and eventually terminate in the amphidial duct. Heterodera glvcines males have the innervation basis for a full complement of sixteen papillae, although surface manifestations are present for only six minute inner labial papillae. In addition, four outer labial and four cephalic receptors terminate beneath the surface, and another two dendrite pairs end further posteriorly beneath the basal plate of the cephalic framework. Papillary receptors which terminate beneath the surface are probably mechanoreceptive, whereas inner labial papillae have pore like openings to the exterior and may be chemoreceptive. Amphids and papillae of H. glycines are fundamentally similar to those of Meloidogyne incognita, although certain striking differences exist. PMID- 19308133 TI - Efficiency of extraction of nematodes by flotation-sieving using molasses and sugar and by elutriation. AB - Blackstrap molasses was studied as an economical substitute for sucrose in the preparation of an extracting solution for removal of nematodes from soil by the flotation-sieving technique. The maximal number of nematodes extracted from soil was obtained with molasses solutions with specific gravity values in the range 1.000-1.073. Results of studies on the relation between size of soil sample and the amount of extracting solution are presented. In paired comparisons, a molasses solution with sp. gr. = 1.100 at 27 C extracted greater numbers of plant parasitic, dorylaimoid, mononchoid, and other soil nematodes than did the standard 1.0 M sucrose solution (sp. gr. = 1.100); the superiority of the molasses solution is attributed to its higher viscosity. The molasses method also was superior or equal in efficiency to the elutriation technique. PMID- 19308134 TI - Interaction of four soybean cultivars with subsoiling and a nematicide. AB - Yields of four soybean, Glycine max, cultivars were increased with subsoiling under the row and application of the nematicide, DBCP i 1,2-dibromo-3 chloropropane) in Tiflon sandy loam heavily infested with the root-knot nematode Meloidogyne incognita. These cultivars represent four maturity groups: very early (V), "Essex', early (VI), "Davis': medium (VII), 'Ransom'; and late (VIII), ' Hutton '. The average increase for the four cullivars was about the same for subsoiling or DBCP. When the treatmcnts were used together, the increase was greater than when either was used alone, but the effects were not additive. Increased yields were obtained with subsoiling and DBCP for the most nematode resistant cultivar, Hutton, as well as for the most susccptiblc, Davis. Subsoiling reduced root-knot galling in nonfumigated plots but did not affect it in fumigated plots. On 12 September, M. incognita larvae were most numerous at the 0- to 20 cm depth, intermediate at 20 to 33 cm depth and least numerous at 33 to 46 cm depth, Subsoiling did not affect larval populations at the three levels. PMID- 19308135 TI - Autoradiography of Developing Syncytia in Cotton Roots Infected with Meloidogyne incognita. AB - Cotton (Gossypium hirsutum) seedlings, uniformly infected with Meloidogyne incognita, were exposed for periods of 1-15 days to a nutrient solution containing tritium-labelled thymidine. Syncytium formation began with the amalgamation of cells near the nematode head, and was followed by synchronized mitoses of the nuclei which had been incorporated into a single cell. Syncytial nuclei synthesized DNA in roots harvested 3, 6, 9, 12, and 15 days after inoculation. Seedlings transferred from unlabelled to labelled nutrient solution 9 days after inoculation, and grown for 6 more days, contained some syncytial nuclei which did not become labelled. Giant-cell nuclei increased in size and, in many cases, all nuclei in one giant cell of a set showed active DNA synthesis at about the time the nematode molted to the adult stage. PMID- 19308136 TI - Scanning electron micrographs of the anterior region of some species of tylenchoidea (tylenchida: nematoda). AB - Micrographs of the anterior region of 42 species in 36 genera of Tylenchoidea obtained with a scanning electron microscope are presented. Greater detail, depth of focus, and structures not previously seen with the light microscope have been obtained in this study. Some of the implications of the morphology of the face view on the classification of the rlylenchoidca are discussed. PMID- 19308137 TI - Marigold, Castor Bean, and Chrysanthemum as Controls of Meloidogyne incognita and Pratylenchus alleni. AB - Root and soil populations of Meloidogyne incognita were significantly fewer from marigold, castor bean, and chrysanthemum than from tomato roots and soil, but not from fallow soil. Root populations of Pratvlenchus alleni were significantly fewer from marigold, castor bean, and chrysanthemum than from tomato: marigold had the fewest. Root populations of M. incognita and P. alleni from tomato simultaneously cultivated with marigold, castor bean, and chrysanthemum were significantly fewer than from tomato cultivated alone. Aborted giant cells and dead M. incognita (larvae and females) were observed in roots of marigold and castor bean, but not in chrysanthemum or tomato. Significantly more males than females occurred in castor bean roots. lnfcction sites of P. alleni appeared normal in all hosts. Thin-layer and column chromatography of alcoholic extracts from castor bean revealed no nematicidal thiophenc derivatives. PMID- 19308139 TI - Pathogenicity of Ditylenchus dipsaci to Sainfoin (Onobrychis viciaefolia Scop.). PMID- 19308138 TI - Population dynamics of plant nematodes as affected by combinations of fallow and cropping sequence. PMID- 19308140 TI - Temperature-induced phase transitions in nematode lipids and their influence on respiration. AB - Temperature-induced phase transitions estimated by electron spin resonance (ESR) technique were ohscrved in the lipids of several nematode species. In both Meloidogyne javanica and Caenorhahditis elegans, there was a phase transition in their phospholipids from a liquid-crystalline state to a solid gel state at about 10 C. Aphelenchus avenae also had a phase transition, but at about 20 C. With this species, the spin-label motion parameters indicated the transition was from the liquid-crystalline state below 20 C to a more liquid or disordered state above 20 C. Anguina tritici and Meloidogyne hapla, in contrast, had no phase transitions over the entire temperature range studied. Each phase transition detected by ESR was reflected in the respiratory rates of the nematodes, and the temperature of the transition coincides with the environmental adaptation of these species. PMID- 19308141 TI - Population Dynamics of Heterodera schachtii on Tomato and Sugar Beet. AB - Experiments showed that development of male and female Heterodera schachtii on tomato and sugarbeet are disproportionately influenced by the nematode inoculum level and root size, which together determine the density of invading larvae. Slight overcrowding favored development of males over females, whereas severe overcrowding equally affected development of males and females. Differential population changes of host-selected races on tested cultivars was attributable to selective development of male and female nematodes. PMID- 19308142 TI - The Incorporation of Photosynthates by Meloidogyne javanica. AB - The root-knot nematode, Meloidogyne javanica, incorporated (1)C from its host after exposure of the plant to (1)CO. This uptake was relatively slow and was not detected in nematodes exposed to a labelled plant for periods of 2 and 4 h, but was after 24 h. Nematodes were grown in plants previously infected at weekly intervals to provide animals at various stages of growth. Plants were harvested 24 h after exposure to the label and the rate of incorporation per unit area of nematode was measured. This rate was found to be related to the nematode's physiological age and reached its peak at the time egg-laying commenced, after which it started to decline. The results support the hypothesis that the nematode functions as a metabolic sink. PMID- 19308143 TI - Pathogenicity of Criconemoides xenoplax to Prune and Plum Rootstocks. AB - Elimination of Criconemoides xenoplax from a prune orchard soil by fumigation with ethylene dibromide at the rate of 42 muliter/liter of soil (equivalent to about 13 gal/acre) improved the growth of Myrobalan plum, Addition of this nematode to Myrobalan seedlings or young 'Marianna 2624' plants propagated from cuttings resulted in destruction of cortical root tissue, darkening of roots, alteration of water stress, lowering of nutrient levels in leaves, and reduction in plant weight. C. xenoplax increased on all nine Prunus cerasifera varieties and hybrids tested, including those used commonly as rootstocks for prunes and plums. Rhizoctonia solani isolated from Myrobalan seedlings infected with C. xenoplax caused lesions on the hypocotyls of young Myrobalan seedlings in the laboratory, but had no effect on older seedlings in the greenhouse, and did not alter the effect of C. xenoplax. PMID- 19308144 TI - A semiquantitative method for enumerating and observing parasites and predators of soil nematodes. AB - A laboratory method was developed to count and observe antagonists of soil nematodes and simulate their relationships in the soil. A 10- to 25-cc soil sample is suspended in water and washed through a series of small standard sieves. Residues are washed into a small beaker and collected on a 24-mm filter paper disk in a filter holder under vacuum. The disk is placed on corn meal agar in a petri dish. Microfauna and flora present in the sample colonize the organic matter on the disk and move onto and into the agar where they can be observed easily. Distinct successions of organisms usually occur and within 6-18 days or more, parasites and predators of nematodes are often abundant, especially nematode-trapping fungi. Counting predation events and parasitized nematodes in replicate dishes after specific incubation periods allows quantitative comparisons between soil samples. The method has distinct advantages over others for enumerating organisms which attack nematodes. PMID- 19308145 TI - Some Ultrastructural Changes Induced in Resistant and Susceptible Soybean Roots Following Infection by Rotylenchulus reniformis. AB - A developmental electron microscopic study of the parasitism of Rolylenchulus reniforrnis in resistant 'Peking' and susceptible 'Lee' soybeans was made during a 21-day period under controlled conditions. Within 2 days of inoculation, the nematode had penetrated the cortical cells to the endodermis where it inserted its stylet, secreted and initiated syncytial formation and cell hypertrophy. Syncytia primarily involved pericycle tissues and, to a lesser extent, xylem parenchyma and endodermis. When identifiable, the cell into which the nematode stylet was inserted to initiate syncytial development was endodermal. Susceptible tissues exhibited two basic phases of development during this infection period: (i) an initial phase represented by partial cell wail lysis and separation; and (ii) an anabolic phase, characterized by organelle proliferation and development accompanied by secondary wall deposits, which provided nutrition for sessile female development. The resistant or hypersensitive reaction (HR) lacked the anabolic phase found in the susceptible reaction, and was characterized by an extension and usually accelerated type of Iysis found in the first phase of the syncytial development. The HR was usually very evident 4 days after inoculation, and could be identified by an almost complete lysis of the cell walls and cytoplasm. The possibility that the initial cell of the developing syncytium or "prosyncyte" may influence a susceptible or resistant reaction is discussed. Successive stages of cell wall dissolution and the deposition of secondary cell walls are described. PMID- 19308146 TI - Effect of Meloidogyne incognita on Selected Forest Tree Species. AB - Four or five growth stages of 14 forest tree species were tested for susceptibility to Meloidogyne incognita at five inoculum levels. Responses ranged from the highly susceptible 'China fir' to immune 'Taiwania'. Even highly susceptible species became increasingly tolerant at later growth stages, thus root-knot appears to be a greater problem in nurseries than in established forests. Heavily suberized cells which restricted nematode development was the predominant host response in Norway spruce, and in the jack, scotch, and Virginia pines. Adult females in jack and scotch pine, which elicited a minimum of suberized tissue, were found adjacent to infection sites showing maximum suberization which indicates that resistance can be highly localized and variable within an individual host. A few gravid females, but no giant cells, were observed in these two species. PMID- 19308147 TI - Pathogenicity and Histopathology of Rotylenchulus reniformis Infecting Cantaloup. AB - Rotylenchulus reniformis was pathogenic to cantaloup (Cucumis melo 'Perlita') under greenhouse conditions. These findings confirm field symptoms of cantaloup infected with R. reniformis. Histopathological studies show that the nematode penetrates the cortex perpendicular to the vascular system and comes to rest with the head against the endodermis in young roots. Feeding stimulated the pericycle to either side of the endodermal feeding cell and caused cell hypertrophy with enlargement of the nucleoli and granular thickening of the cytoplasm. In older roots where the endodermis had collapsed, the nematode fed directly into the pericycle and caused similar symptoms. Nematode development was more rapid at 27 C than at 21 C. PMID- 19308148 TI - Feeding, Egg-Laying, and Embryology of the Columbia Lance Nematode, Hoplolaimus columbus. AB - Feeding and egg-laying of Hoplolaimus columbus were observed on excised alfalfa (cultivar 'DuPuits') root cultures in 1.0% nutrient agar. Feeding was ectoparasitic on cortical cells in the maturation zone of the root. Oocytes were first observed in the anterior ovary of the feeding female 8-9 days after feeding began. Globular secretions emanating from the vagina and vulva preceded migration of the posterior-most oocyte of the anterior gonad into the columella. The egg shell was formed within 8 h, and the egg was laid within 12-24 h. The eggs differed from those of other plant-parasitic nematodes in having a stalk on the distal end. The average time required from egg-laying to hatching was 12 days in water and in alfalfa root cultures. PMID- 19308149 TI - Crop rotation and herbicide effects on population densities of plant-parasitic nematodes. AB - The influence of herbicides and mono- and multicropping sequences on population densities of nematode species common in corn, cotton, peanut, and soybean fields in the southeastern United States was studied for 4 years. Each experimental plot was sampled at monthly intervals. The application of herbicides did not significantly affect nematode population densities. Meloidogyne incognita and Trichodorus christiei increased rapidly on corn and cotton, but were suppressed by peanut and soybean. More Pratylenchus spp. occurred on corn and soybean than on cotton and peanut. Criconemoides ornatus increased rapidly on corn and peanut, but was suppressed by cotton and soybean. Helicotylenchus dihystera was more numerous on cotton and soybean than on corn and peanut. Numbers of Xiphinema americanum remained low on all crops. The peanut sequence was the most effective monocrop system for suppressing most nematode species. Multi-crop systems, corn peanut-cotton-soybean and cotton-soybean-corn-peanut, were equally effective in suppressing nematode densities. PMID- 19308150 TI - Longidorus breviannulatus n. sp. (Nematoda: Longidoridae) Associated with Stunted Corn in Iowa. AB - Longidorus breviannulatus n. sp. was associated with stunted corn in Iowa. The nematode has wide bilobed amphidial pouches, a guiding ring 21-26 mum from the anterior end of the body, an odontostyle 81-88 mum long, and a spear extension 28 45 mum long. The most frequent collections and the greatest numbers of the nematode in the field occurred in sandy soils. Nematodes increased in greenhouse culture from an initial population of 100 to 4,120 individuals in 322 days. PMID- 19308151 TI - Factors Affecting the Control of Rotylenchulus reniformis with Electromagnetic Energy. AB - The reniform nematode Rotylenchulus reniformis was reduced in the upper 10 cm of soil with application of UHF electromagnetic energy. Bioassay of treated soil indicated no delayed effect on the population from the treatment. The population was significantly reduced by hot water treatments at 40 C for 10 min, and at 45 C for 5 and 10 min, 50 C and above killed all nematodes. Data were inconclusive as to whether the effect of UHF electromagnetic energy was thermal or nonthermal. PMID- 19308152 TI - Body Wall Fine Structure of the Anterior Region of Meloidogyne incognita and Heterodera glycines Males. AB - The body wall fine structure including the cuticle, hypodermis, and somatic muscles is similar in males of Meloidogyne incognita and Heterodera glycines. The cuticle can be regarded as basically three-layered in both species, but is much thicker in M. incognita than in H. glycines, and differences occur in surface markings. The chordal and interchordal hypodermis is syncytial. Hypodermal tissue pervades the lip region, and lines the stomatal cavity and stylet shaft. Various organelles and structures, some previously undescribed, are concentrated in the chords. Their possible role in lipid metabolism is considered, as well as the probable function of the hypodermis in fornlation of the cephalic framework and stylet. The interchordal hypodermis which encloses peripheral nerves, is periodically transversed by bundles of fibrils which are homologous with the subcuticular striation previously observed in the light microscope. The somatic musculature is meromyarian, and the muscle cells are of the platymyarian type with I, A, and H bands, but without Z bands or T tubules. Thin dense bands are present in the H bands, and appear to be associated with sarcoplasmic reticulum. PMID- 19308153 TI - Correlations of field populations of nematodes with crop growth responses for determining relative involvement of species. AB - Treated and nontreated field plots were assayed, and the population density of each plant-parasitic nematode present was compared with crop growth and yield and with the population densities of other nematode species in the same plots. The strongest correlations between nematode population densities and growth responses occurred when soil assays for nematodes were made 55-73 days after planting. Belonolairnus longicaudatus was the most damaging parasite on peanut, Arachis hypogaea, as evidenced by high negative correlations between population densities and plant growth responses. Criconemoides ornatus, Meloidogyne hapla, Helicotylenchus dihystera, Trichodorus christie, Tylenchorhynchus claytoni, and Pratylenchus brachyurus were involved to varying degrees, depending on previous crop and initial densities of these nematodes. Hoplolaimus galeatus and Xiphinema americanum did not appear to affect crop response. The negative correlation of Trichodorus christiei to yield of soybean, Glycine max, was higher than that of Belonolairnus longicaudatus, although both contributed to yield losses. Similar correlation analyses showed that apparent antagonistic or synergistic population density relationships among nematodes under field conditions depend on the time of sampling and the composition of the nematode community under study. PMID- 19308154 TI - Freezing and Storing Ditylenchus dipsaci in Liquid Nitrogen. AB - After 18 months of storage at -150 C, some larvae of Ditylenchus dipsaci, which had been treated in a 7.5% solution of dimethyl sulphoxide and cooled to -25 C before storage, were still viable on thawing. Some survivors penetrated and developed normally in stems of alfalfa seedlings. Tests showed that active larvae could be frozen directly, thus eliminating the need to use the quiescent stage of this nematode previously thought necessary for successful storage at cryogenic temperatures. The method described is suitable for long-term storage of D. dipsaci and may, with slight modifications, be used to preserve other plant parasitic nematodes. PMID- 19308155 TI - The Use of Density-Gradient Centrifugation for the Purification of Eggs of Meloidogyne spp. PMID- 19308156 TI - Endotokia matricida in a Xiphinema sp. PMID- 19308157 TI - Penetration and development of the mermithid nematode Reesimermis nielseni in eighteen species of mosquitoes. AB - The susceptibility of 18 species of mosquitoes to the infective stage of the mermithid nematode Reesimermis nielseni was compared to that of Culex pipiens quinquefasciatus. Thirteen species were more susceptible, with three Anopheles species and Culiseta inornata the most susceptible. Aedes triseriatus, Culex territans, and Psorophora ferox were highly resistant. Resistance to R. nielseni appeared to be behavioral, physical, or physiological, and some host species exhibited one or more types of defense mechanisms. No noticeable differences were apparent in the degree of susceptibility of a native-and a laboratory strain of C. p. quinquqfasciatus to R. nielseni. PMID- 19308158 TI - Development and fecundity of Reesimermis nielseni, a nematode parasite of mosquitoes. AB - Maturation of the mermithid nematode Reesimermis nielseni to the adult stage began by the tenth day after emergence of the nematodes from their hosts at ambient temperatures (24-27 C). Most postparasitic males and females reached the adult stage after 50 and 70 days, respectively. The first females exhibiting egg development and oviposition were observed 25-30 days after emergence, but some oviposition was still taking place 150 days later. Reesimermis nielseni laid an average 2,480 eggs per female over an 18-day oviposition period. A majority of the mature eggs hatched within 7 h after the cultures were flooded. The preparasites are short-lived, and only a few were able to infect exposed hosts after 72 h. PMID- 19308159 TI - Behavior of aldicarb in soil relative to control of Heterodera schachtii. AB - The adsorption characteristics of two soils for aldicarb sulfoxide were similar to that described by the Freundlich equation, The adsorption constant for the Holtville clay was 3.3, and that of the Buren silt loam, 0.34. Planting beds in a field of Holtville clay and another of Buren silt loam were side-dressed at 25 kg and 50 kg/ha 10% aldicarb (Temik 10G). Comparison of field measurements of aldicarb concentrations with previous laboratory determinations of aldicarb effects on Heterodera schachtii allowed predictions of soil zones in which hatching, infectivity, and orientation of males to females would be affected. Aldicarb in the soil water of Holtville clay sufficient to interfere with male orientation extended through most of the bed profile to a depth of 46 cm 1 week after the first irrigation. Orientation could be affected in only the top 20 cm of the bed 37 days after treatment and application of 712 mm of irrigation water. In Buren silt loam, disorientation of males was estimated to occur throughout the bed 42 days after treatment and 600 mm irrigation water. Aldicarb persisted in extensive areas of the bed at concentrations sufficient to prevent infection. In small areas of the profile, aldicarb sufficient to inhibit hatching persisted. Amounts of aldicarb in soil water samples obtained directly from beds agreed well with those from the analysis of the air dried soil samples. PMID- 19308160 TI - Effects of aldicarb on the behavior of Heterodera schachtii and Meloidogyne javanica. AB - The toxic effects of sublethal concentrations of aldicarb were studied on eggs and second-stage larvae and males of Heterodera schachtii and second-stage larvae only of Meloidogyne javanica in a quartz sand substrate. Aldicarb was more toxic to eggs of H. schachtii than to those of M. javanica. Complete suppression of hatching occurred between 0.48 and 4.8 microg/ml aldicarb for H. schachtii whereas 100% inhibition of hatch of M. javanica occurred between 4.8 and 48.0 microg/ml. M. javanica hatch was stimulated at 0.48 microg/ml aldicarb. Migration of second-stage larvae of H. schachtii and M. javanica in sand columns was inhibited under continuous exposure to 1 microg/ml aldicarb. Infection of sugarbeet and tomato seedlings by larvae was inhibited at 1 microg/ml. H. schachtii males failed to migrate toward nubile females at 0.01 microg/ml aldicarb. This was partially confirmed in a field study in which adding aldicarb to soil resulted in fewer females being fertilized. PMID- 19308161 TI - Effects of soil temperatures and inoculum levels of Meloidogyne incognita and Rhizoctonia solani on seedling disease of cotton. AB - Soreshin of cotton was more severe from combined infections of Rhizoctonia solani and Meloidogyne incognita than from either organism alone, when both critical soil temperature and inoculum concentrations were present. Optimum soil temperatures for disease development from combined infections were 18 and 21 C. Either 2,500 or 5,000 M. incognita larvae per plant, combined with R. solani, increased seedling disease severity over that caused by R. solani alone. When 100 or 500 larvae per plant were added with R. solani, disease severity did not change. Disease severity increased with the highest level of R. solani inoculum either alone or combined with M. incognita. PMID- 19308162 TI - Effects of soil texture on the interaction between Rhizoctonia solani and Meloidogyne incognita on cotton seedlings. AB - Soils containing 60, 75, and 90% coarse particles (sand plus coarse silt) were prepared by dilution of a field soil with 246microm (60-mesh) silica sand. As the coarse-particle content of the soils increased, the synergistic interaction between Meloidogyne incognita and Rhizoctonia solani on cotton seedlings increased. Increasing the coarse-particle content of the soil also increased damage from the nematode alone and slightly increased soreshin damage from R. solani alone. PMID- 19308163 TI - Parasitism of Nonhost Cultivars by Ditylenchus dipsaci. AB - The alfalfa race of Ditylenchus dipsaci parasitized and caused characteristic symptoms on nonhost seedlings of sweet clover, onion, tomato, sugarbeet, and wheat in controlled growth-chamber studies. Although the nematode was unable to reproduce on any of the cultivars, it caused plant mortality ranging from 20% on sugarbeet and tomato to 100% on onion. PMID- 19308164 TI - Effects of Temperature on the Fine-Structural Responses in the Hypocotyl Region of Alfalfa Lines to Ditylenchus dipsaci. AB - Fine-structural analyses were made of the response of host tissue, Medicago sativa L. 'Ranger' and 'Lahontan', to infection by the stem nematode, Ditylenchus dipsaci (Kuhn) Filipjev. Seedlings were grown at 15 and 25 C, and hypocotyl regions were sampled 1,3, or 7 days after inoculation. Electron micrographs of infected alfalfa tissue indicated that the same types of damage were inflicted on Lahontan (stem-nematode-tolerant)and Ranger (stem-nematode-susceptible). Only the infection rate and degree of damage differed between lines and temperatures, with the greater injury occurring at the higher temperatures. After 3 and 7 days of infection, the symptoms observed were: swollen and broken endoplasmic reticulum (ER), distended and broken chloroplasts, loss of nuclear material, and bulging and rupturing of nuclear envelopes. Cells with infected cytoplasm contained more ER, ribosomes, vesicles, and Golgi apparatuses, suggesting increased metabolic activities. Lobing nuclei were observed in all samples. Lipid contents varied with temperature in I-day-old seedlings. At 15 and 25 C, electron-dense substrances were commonly found along the tonoplast, and on the cell wall. Also, some cells with enlarged ER were noted in the noninfected controls at these temperatures. PMID- 19308165 TI - Tylenchocriconema alleni n. g., n. sp. from Guatemala (Tylenchocriconematidae n. fam.; Tylenchocriconematidae n. superfam.; Nematoda). AB - Tylenchocriconema alleni n.g.n.sp, is described from soil about roots of a bromeliad from Guatemala City, Guatemala. It has characteristics of both the Tylenchoidea and Criconematoidea. Tylenchocriconematidae, n. fam., and Tylenchocriconematoidea, n. superfam., are proposed to express the relationship of this species to other taxa. Tylenchulidae and Paratylenchidae are considered closely related, and Tylenchulidoidea, n. rank, is proposed to indicate their relationship. PMID- 19308166 TI - Penetration of 1,2-dibromo-3-chloropropane in a Florida soil. AB - Gas-liquid chromatography was used to detect movement of the nematicide, 1,2 dibromo-3-chloropropane (DBCP), in soil columns containing top- and subsoil of Astatula fine sand. Topsoil contained 1.4-1.6% organic matter and subsoil 0.20 0.25%. DBCP was applied at various rates as aqueous drenches. Depth of penetration was controlled by organic matter in topsoil and varied with the amount of water applied. Maximum DBCP penetration after 14 days was 28 cm; maximum water infiltration, 115 cm. Maximum depth of penetration was obtained with a water emulsion of 30 mug/ml of DBCP applied in 15 cm of water. DBCP applied in 5 cm of water to soils containing 2.0% and 0.125% organic matter penetrated 6 cm and 60 cm, respectively. PMID- 19308167 TI - Meloidogyne incognita-Induced Changes in Cell Permeability of Galled Roots. AB - Electrolyte leakage of Meloidogyne incognita-infected and healthy tomato roots was compared by conductivity measurements, and by compartmental analysis using Rb. Conductivity measurements suggested difference in electrolyte loss from healthy and galled roots. On a percentage basis, a greater rate of efflux occurred for healthy plants, but galled roots contain more electrolytes and may show a larger net loss. Compartmental analysis indicated that: (i) the longer half-time for Rb loss from vacuoles of galled root cells could indicate either a greater vacuolar content or decreased tonoplast permeability, (ii) the shorter half-time for loss Rb from the cytoplasm of galled root cells could reflect either a reduced cytoplasmic content or an increased plasma membrane permeability, and (iii) in split-root plants, the permeability of the tonoplast and the plasma membrane of cells in nongalled roots is increased by nematode infection on the other half of the root system. Thus, a mechanism for mobilizing minerals to the infection site is proposed. PMID- 19308168 TI - Rearing migratory endoparasitic nematodes in citrus callus and roots produced from citrus leaves. AB - Radopholus similis and Pratylenchus coffeae were reared on callus and roots developed from citrus leaves. Callus formed best when leaf petioles were immersed in Astatula fine sand and the leaves were sprayed daily with 4 ppm 2,4-D solution and maintained at 25 or 30 C. The nematodes completed one generation in 20 days at 25 C. Highest populations of R. similis (1,127) occurred after 50 days, and the highest for P. coffeae (619) after 70 days. Leaf-callus cultures from R. similis-resistant citrus rootstocks showed the same degree of infection as susceptible rough lemon callus after 30 days. PMID- 19308169 TI - Four new species of the genus hemicriconemoides (nematoda:criconematidae). AB - Four new species of the genus Hemicriconemoides (H. californianus n.sp., H. taiwanensis n.sp., H. annulatus n. sp., and H. nitida n.sp.) are described. The range of total length of H. mangiferae is increased on the basis of specimens collected in Israel, Observations on H. mangiferae and H. litchi support the validity of H. litchi as distinct from H. mangiferae. PMID- 19308170 TI - The Effects of DBCP on Citrus Root Nematode and Citrus Growth and Yield in Iraq. AB - In lraq, treatment of producing citrus trees with a 75% emulsifiable formulation of 1,2-dibromo-3-chloropropane (DBCP), applied in irrigation water at rates of 66 88 kg (a.i.)/hectare, gave excellent control of the citrus nematode (Tylenchulus semipenetrans) greatly improved tree vigor, and increased the fruit yield for at least 3 years after treatment. Applications made during the spring gave much better results than those made in the fall. PMID- 19308171 TI - Revision of the Genus Paratylenchus Micoletzky, 1922 and Descriptions of New Species. Part II of Three Parts. AB - Part II covers species with average female stylet length of 22-38 mum. Seven new species are described and further observations are given on 12 other species. A key to the species covered in Parts I and II is included. Paratylenchus curvitatus van der Linde, 1938, is transferred to species inquirendae. PMID- 19308172 TI - Relationship of grapevine yield and growth to nematode densities. AB - Yield, growth, and vigor of individual grape vines were correlated with nematode population densities in a series of California vineyards. In a Hanford sandy loam soil, Xiphinema americanum densities showed negative correlations with yield, growth, and vigor of vines. When vines were categorized according to vigor, X. americanurn densities had little relationship to yield of high-vigor vines, but were negatively correlated with yield of low-vigor vines. Densities of Paratylenchus harnatus were positively correlated with yield, growth, and vigor of vines. Correlations between Meloidogyne spp. densities and vine performance were variable, even when the vines were separated according to soil type and plant vigor. Densities of Meloidogyne spp. populations were generally higher on coarser-textured, sandy soils and the vines were less vigorous there. Densities of P. hamatus were greater in fine-textured soils. PMID- 19308173 TI - In-Vitro and In-Vivo Effects of Aldicarb on Survival and Development of Heterodera schachtii. AB - Aqueous solutions of 5-500 mug/ml aldicarb inhibited hatching of Heterodera schachtii. Addition of hatching agents, zinc chloride, or sugarbeet root diffusate, to the aldicarb solutions did not decrease the inhibition of hatching. When cysts were removed from the aldicarb solufions and then treated for 4 wk in sugarbeet root diffusate, larvae hatched and emerged. Treatments of newly hatched larvae of H. schachtii with 5-100 mug/ml aldicarb depressed later development of larvae on sugarbeet (Beta vulgaris). Similar treatments with aldicarb sulfoxide had less effect on larval development, and aldicarb sulfone had no effect. Numbers of treated larvae that survived and developed were inversely proportional to concentration (0.1-5.0 mug/ml) and duration (0-14 days) of aldicarb treatments. Development of H. schachtii on sugarbeet grown in aldicarb-treated soil was inversely proportional to the concentration of aldicarb in the tested range of 0.75 - 3.0 mug aldicarb/g of soil. Transfer of nematode-infected plants to soil with aldicarb retarded nematode development, whereas transfer of plants first grownin treated soil to nematode-infested soil only slightly suppressed nematode development. Development of H. schachtii was inhibited in slices of storage roots of table beet (B. vulgaris), sugarbeet and turnip, (Brassica rapa), that had grown in soil treated with aldicarb. PMID- 19308175 TI - Applicator for liquid nematicides in research plots. PMID- 19308174 TI - A Note on Xiphinema ensiculiferum. PMID- 19308176 TI - Variation of Tail Shape of Cylindrocorpus curzii (Nematoda:Cylindrocorporidae). PMID- 19308177 TI - Influence of Meloidogyne incognita on the content of amino acids and nicotine in tobacco grown under gnotobiotic conditions. AB - Seedlings of Meloidogyne incognita-resistant (N.C. 95) and -susceptible (McNair 30) tobacco cultivars were grown aseptically for 55 days inside isolator chambers in autoclaved soil infested with 0 or 3,000 axenized eggs of M. incognita per 500 cc of soil. Healthy and infected plants were compared. Dry root weights of infected plants of resistant and susceptible cultivars were 16% and 84%, respectively, less than the controls. Sixteen amino acids, including those precursors for nicotine, and nicotine, increased significantly in infected roots of both cultivars. Increases in amino acids in infected roots ranged from 28% for valine to 103% for tyrosine in the resistant N.C. 95, and from 30% for leucine to 148% for tyrosine in lhe susceptible McNair 30. Nicotine content (dry weight basis) increased 42% and 62% in infected roots of resistant and susceptible cultivars, respectively. Nematode infection increased nicotine by 112% in leaves of N.C. 95, and decreased it by 56% in leaves of McNair 30. Root damage by M. incognita probably decreased nicotine movement into leaves of McNair 30. In N.C. 95, nicotine movement into leaves apparently was not adversely affected due to lack of significant root damage. PMID- 19308178 TI - The pin nematode, Paratylenchus neoamblycephalus, on Myrobalan plum and other hosts. AB - Elimination of Paratylenchus neoamblvcephalus from soil by fumigation with 1,2 dibromoethane stimulated the growth of Myrobalan seedlings grown in it. Addition of a suspension of P. neoamblycephalus to Myrobalan seedlings inhibited their growth as compared to noninoculated controls. When nematodes were removed from the suspension by settling, and the supernatant liquid was used as inoculum, no stunting occurred. Roots of Myrobalan seedlings inoculated with surface sterilized P. neoamblycephalus were smaller, darker, and had fewer feeder roots than those of noninoculated controls. Nematodes were observed feeding ectoparasitically, but with heads embedded in roots as deep as the cortex. They were associated with small lesions and dead lateral roots. Clusters of nematodes were common at ruptures in the epidermis, and where lateral roots emerged. Limitation of Myrobalan growth by P. neoamblvcephalus was greater at 20 and 27 C than at 30 C, and was not affected by pH over the range 4.5 to 6.5. Rose, apricot, peach, and all selections and hybrids of Prunus cerasffera tested were hosts for P. neoamblrcephalus. The nematode could not be cultured on various herbaceous plants nor on Myrobalan callus tissue. PMID- 19308179 TI - Nematode community structure in desert soils: nematode recovery. AB - The sugar-flotation-sieving (SFS) and Baermann-funnel (BF) methods were compared for nematode extraction efficiency. The SFS method recovered nematodes from more trophic groups whereas greater total numbers of individuals were recovered by BF. In a test to validate the efficiency of SFS, virtually 100% of the nematodes added to desert soil prior to extraction were recovered by four consecutive SFS washings of each soil sample. Estimations of nematode biomass in desert soils based on numbers of nematodes extracted by the two methods were similar unless there was large reserve of eggs in the soil. The biomass of nematodes from a Colorado desert soil was 0.9 g/m(2) as determined by both methods, whereas BF gave 0.17 g/m(2) for nematodes from a Mojave desert soil as compared to 0.9 g/m(2) with SFS. PMID- 19308180 TI - Control of Heterodera schachtii with foliar application of nematicides. AB - Foliar applications of ethyl 4-(methylthio)-m-tolyl isopropylphosphoramidate (phenamiphos) or S-methyl 1-(dimethylcarbamoyl)-N-[(methylcarbamoyl)oxy] thioformimidate (oxamyl) retarded infection of sugarbeets by the sugarbeet nematode, Heterodera schachtii under greenhouse conditions. Maximum nematode control was obtained when treatments were applied previous to, or at the time of, inoculation of plants with the nematode. Consecutive foliar applications inhibited nematode development, with four applications giving greatest inhibition of maturation. A treatment with either phenamiphos or oxamyl at 2,000 microg/ml (ppm) resulted in the greatest increase in plant growth, and 4,000 microg/ml gave the best nematode control. A treatment of 4,000 microg/ml of either phenamiphos or oxamyl was phytotoxic. However, this was due to container confinement of the chemical since phytotoxicity at this rate has not been observed under field conditions. PMID- 19308181 TI - Relationships between the population density of Meloidogyne incognita and growth of tobacco. AB - Seedlings of tobacco cultivars resistant (NC95) and susceptible (McNair 30) to Meloidogyne incognita were grown in 15-cm diameter clay pots containing steamed soil infested with 0, l, 2, 4, 8, 16, 32, and 64 eggs of M. incognita per 1.5 cm(3) soil. Plants were maintained in the greenhouse for 3 weeks, and then transferred to the field for 12 weeks. Growth of tobacco was expressed separately as dry weight of leaves and as plant height. Least squares regression analysis showed that tobacco growth-nematode density interactions are in agreement with Seinhorst's exponential model Y = m + (l-m) czp. Tobacco growth was not affected significantly as nematode density was increased from 0 to tolerance levels, which were approximately 2 and 1 eggs per 1.5 cm(3) soil for the resistant and susceptible cultivars, respectively. As nematode density was increased beyond tolerance level, tobacco growth decreased sharply until a minimum yield was approached. The minimum leaf weights and plant heights of the resistant cultivar at the highest nematode density were greater than those of the susceptible cultivar. PMID- 19308182 TI - Genetic structure of races of Heterodera glycines and inheritance of ability to reproduce on resistant soybeans. AB - Four field populations of Heterodera glycines tested for ability to reproduce on three host differentials were each classified into one of the recognized races. A fifth population represented a new race. Genetic analysis indicated that the designated races are actually field populations that differ from each other primarily in the frequencies of three groups of genes (genes for parasitism) that act quantitatively and control the ability of the nematode to reproduce on resistant P.I. 88788, Pickett, and P.I. 90763 soybeans. Populations of race-3 have none of these genes for parasitism, or they have some in low frequency that results in an index of parasitism of less than 10 on any one of the resistant soybeans. Race-1 has a high frequency of one group of genes that enable it to reproduce on P.I. 88788. Race-2 has two groups of genes for parasitism in high frequency; one for P.I. 88788, and one for Pickett. Based on these findings, it was assumed that race-4 has three groups of genes for parasitism; one for P.I. 88788, one for Pickett, and one for P.I. 90763. Additional races may be recognized when new genes are identified, or when new gene combinations are discovered. The ability to reproduce on P. I. 88788 is inherited independently from the ability to reproduce on Pickett. Although the genetic structure of field populations does not provide a solid foundation for race designation, recognizing races under the present system may be useful when it clearly characterizes the behavior of field populations. Race designations, however, should be regarded as provisional since gene frequencies change with time in response to selection forces and, therefore, the race status of a population may change accordingly. PMID- 19308183 TI - Effects of light intensity and quality effects on reproduction of plant-parasitic nematodes. AB - Growing cotton in a greenhouse with 12-h of supplemental light [8,608 lux (800 ft c) from combination of mercury and Lucalux lamps] resulted in 2 x to > 3 x greater reproduction of Meloidogyne incognita and Belonolaimus longicaudatus as compared to natural light alone. Rate of increase of Hoplolaimus galeatus was affected little in this experiment. In a second experiment under controlled conditions in a phytotron, light source and intensity had greater influence on the reproduction of Heterodera glycines and Pratylenchus penetrans on soybean than on B. longicaudatus. Fluorescent plus incandescent and metal halide light sources resulted in the greatest nematode reproduction. Lucalux lamps resulted in much lower rates of nematode increase than other light sources. Rates of nematode increase on soybean under the different light sources in the phytotron generally were positively related to plant growth. PMID- 19308184 TI - Effect of RotyIenchulus reniformis on reflectance of cotton plant leaves. AB - Differences between light reflectance from leaves of cotton (Gossypiurn hirsutum) plants grown with a low- or no-nematode (Rotylenchulus reniformis) population (nonstressed), and from leaves grown with a high nematode population (stressed) were measured in field and greenhouse experiments. Reflectance was measured spectrophotometrically in the laboratory on single leaves and spectroradiometrically in the field on plant canopies. Nematode-stressed cotton plants were stunted with fewer, smaller, and darker-green leaves than nonstressed plants. Over the 0.5- to 2.5-/microm waveband, stressed leaves had lower reflectance than nonstressed leaves of the same chronological age for both field- and greenhouse-grown plants. Reflectance differences between stressed and nonstressed leaves in the visible (0.5 to 0.75 microm), near-infrared (0.75 to 1.35 mum) and infrared water absorption (1.35 to 2.5 microm) regions were primarily caused by differences in leaf chlorophyll concentration, mesophyll structure, and water content, respectively. Results indicate the potential for remotely sensing nematode-infested plants to distinguish them from normal plants. PMID- 19308185 TI - Rhabditis pellio Schneider (nematoda) from the earthworm, Aporrectodea trapezoides Duges (Annelida). AB - Studies were conducted on the behavior of the nematode, Rhabditis pellio, in the earthworm, Aporrectodea trapezoides, from southern California. Juvenile and adult nematodes were found in the bladders and tubules of the metanephridia of the host. Similar nematodes that entered the coelom were encapsulated and incorporated into multiple capsules ("brown bodies"). It was demonstrated that this host response is an effective defense reaction since dead and dying nematodes, as well as living forms, were found in the capsules. PMID- 19308186 TI - Factors affecting the infection of alfalfa seedlings by Ditylenchus dipsaci. AB - Experiments were conducted to determine the effects of plant confinement, soil type, watering practices, stage of seed germination, inoculum level, and method of applying inoculum on stem nematode (Ditylenchus dipsaci) infection of alfalfa (Medicago sativa) seedlings grown in soil. Results indicated that (i) confining seedlings together with the nematodes in small vials offered no advantage over growing plants in large flats, (ii) a very fine sandy-loam soil was superior to a fine sand for stem-nematode penetration, (iii) nematodes penetrated seedlings more readily if the soil was not watered immediately after planting and inoculation, (iv) germinating seeds with a radicle length of 0.6-1.3 cm had the highest nematode penetration, and (v) highest penetration occurred when the nematodes were placed directly upon germinating seeds. The optimum inoculum level was 50 nematodes per seedling. PMID- 19308188 TI - Comparative fine structure of the stomatal region of males of Meloidogyne incognita and Heterodera glycines. AB - The fine structure of the stylet, "guiding apparatus," anti protractor muscles of males of Meloidogyne incognita and Heterodera glycines is ehtcittated and compared. In both nematodes, the stylet cone is of greater electron density than the shaft. The cone is heterogeneous; shaft material extends into it for most of its length, whereas the shaft proper and knob regions are relatively homogeneous. The stylet lumnen is round throughout its length in H. glycines, but in M. incognita changes from round, posteriorly, to irregular, triradiate, and oblong near the styler tip. The various layers of the lining of the stylet shaft are continuous with those of the cuticular lining of the esophageal lumen. Similarly, cuticular layers of the stomatal lining and vestibule extension are continuous with the body wall cuticle. The stylet "guiding apparatus" is formed by linings of the stomatal opening, vestibule, and vestibule extension, together with transversely folded membranes, which extend further posteriorly and attach near the junction of shaft and cone. Noncontractile regions of the three stylet protractor muscles originate in the esophagus. Contractile portions extend anteriorly from the stylet knobs and branch into a total of 10 elements that attach near the basal ring of the cephalic framework and the vestibule extension. A second, noncontractile region is present at the anteriormost part of the protractor muscles in H. glycines. PMID- 19308189 TI - Influence of Pratylenchus vulnus and Meloidogyne hapla on the growth of rootstocks of rose. AB - Pratylenchus vulnus is involved in a desease of Rosa noisettiana 'Manetti' rose rootstock characterized by darkening of roots, death of feeder roots, and stunting of entire plants. The disease is more severe when plants are grown in silt loam soil than when they are grown in sandy loam soil. The nematodes reproduce best in silt loam soil at 20 C. Meloidogyne hapla did not affect the growh of Manetti. Rosa sp. 'Dr. Huey', Manetti, and R. odorata rose rootstocks were found to be goos hosts for P. vulnus whereas R. multiflora was less suitable. M. hapla reproduced well on R. odorata, Dr. Huey, and R. multiflora, but not on Manetti. PMID- 19308190 TI - Improved methods of hatching Heterodera schachtii larvae for screening chemicals. AB - The rate of hatching of Heterodera schachtii larvae was greatly increased by placing cysts in sieves enclosed by small disposable cups. An apparatus that permitted rapid storage of second-stage larvae at 10 C prolonged the viability of the larvae. PMID- 19308191 TI - Susceptibility of Japanese holly to Criconemoides xenoplax, Tylenchorhynchus claytoni, and certain other plant-parasitic nematodes. AB - Three cultivars of Ilex crenata: 'Helleri', 'Convexa', and 'Rotundifolia' were inoculated with either Criconemoides xenoplax, Helicotylenchus dihystera, Hoplolaimus galeatus, Trichodorus cltristiei, or Tylenchorhynchus claytoni at 0, 200 (low), or 2,000 (high) nematodes/ 15-cm diam pot. Plants were kept in the greenhouse 10 mo prior to transplanting into 2.25 m(2) field plots. Helleri was severely stunted by C. xenoplax. Criconemoides xenoplax and T. claytoni caused lower plant vigor and top weights of Rotundifolia after 3 years. Above-ground symptoms included stunting, chlorosis, and leaf drop. Convexa was not susceptible to the nematodes tested. Low and high initial populations of the five nematodes tended to reach equilibria over the 3-year sampling period. Helicotylenchus dihystera and C. xenoplax occurred in the greatest densities regardless of host. None of the test plants were damaged by H. dihystera. Convexa was the least suitable host for nematode reproduction. Hoplolaimus galeatus, which was originally isolated from cotton, failed to reproduce or survive on any plant tested. Nematode densities over the 3-year sampling period did not always lit a linear regression model. PMID- 19308192 TI - Distribution patterns of some criconematinae in different forest associations. AB - A total of 243 samples from Hemlock-Hardwood, Boreal Forest, and Alpine Tundra associations of New York, Vermont, New Hampshire, and Maine were analyzed for species of Bakernema, Criconema, and Criconemoides and for selected edaphic factors. The Hemlock-Hardwood formation contained 13 species of these genera, but the Boreal Forest and Alpine Tundra contained only Criconema menzeli and Criconemoides sphagni. Criconemoides axeste, C. rusticum, and C. xenoplax were associated primarily with mineral soils that have high pH, low moisture after drainage, and organic matter content of less than 15%. Criconemoides sphagni was associated with organic soils that had low pH, high moisture after drainage, and organic matter content greater than 15%. PMID- 19308193 TI - Persistence of activity of oxamyl against Heterodera schachtii on cabbage. AB - The duration of effectiveness of a foliar spray of oxamyl against Heterodera schachtii and the location of the protective effect were determined by applying a foliar spray at 0.04 kg (a.i.)/100 liters of water to cabbage seedlings. Oxamyl, or a metabolite of oxamyl, apparently is translocated to anti becomes prolective in the root within 7 days. Between 7 and 14 days, the location of the protection shifts from within the root In the root surface or rhizosphere. The chemical remains active for at least 21 days unless it is removed from the root or rhizosphere by washing with water. PMID- 19308194 TI - Efficacy of oxamyl against Heterodera schachtii on cabbage. AB - The efficacy of oxamyl in controlling Heterodera schachtii on cabbage was determined by applying various contbinations of soil drenches at 6.7 kg (a.i.)/ha and foliar sprays at 0.04 kg (a.i.)/100 liters of water to cabbage seedlings. Pretransplant drenches provided some control of H. schachtii over a 13-week period. A single foliar spray of oxamyl 1 week before transplanting apparently prevented penetration of H. schachtii larvae; post-transplant sprays were relatively ineffective. A pretransplant or transplant drench combined with a foliar application 2 weeks after transplanting provided the most effective control. The effectiveness of drenches plus post-transplant sprays is probably due to the spray augmenting the action of the drench in inhibiting the development of larvae after penelration. PMID- 19308195 TI - Self-interactions of Meloidogyne hapla and of Heterodera schachtii on beta vulgaris. AB - Double inoculations of sugar beet with larvae of Meloidogyne hapla resulted in a higher galling incidence in only one treatment than did a single inoculation using the same number of larvae. Double inoculations with larvae of Heterodera schachtii, however, resulted in three- to five-fold more cysts in most cases than did single inoculations using the same number of larvae. In general, plants died more quickly after double inoculations than after single inoculations of the same total number of either nematode. Ratios of total soluble carbohydrates to reducing carbohydrates were lower in multiple inoculated treatments than in other treatments. Plants infected with M. hapla had lower quantities of B, K, and P in leaf tissue than noninoculated plants, but no differences were correlated with type of inoculation. Plants inoculated with H. schachtii had lower quantities of B, K, and Mg than noninoculated plants. Also, quantities of Mn, Cu, and Zn were much lower in plants inoculated twice with H. schachtii larvae than in plants inoculated with the same total number of larvae in a single dose. PMID- 19308196 TI - Influence of nematodes and light sources on growth and nodulation of soybean. AB - The influence of nematodes on nodulation of soybean varied according to their modes of parasitism. In the greenhouse, nodule formation was stimulated by the endoparasites, Meloidogyne hapla and Pratylenchus penetrans, but was inhibited slightly by the ectoparasite, Belonolaimus longicaudatus. In an experiment under controlled conditions in a phytotron, Heterodera glycines severely inhibited nodule formation, whereas plants inoculated with B. longicaudatus and P. penetrans had more nodules per g root than nematode-free plants. Nitrogen-fixing capacity, however, was inhibited by all three nematode species. Different light sources used in the phytotron experiment also influenced growth and nodulation of soybean. A fluorescent plus incandescent light regime resulted in plants with the greatest shoot weight, pod number, and nodules per g root. Plants grown under Lucalox lamps had excessive stem elongation. PMID- 19308197 TI - The genus Malenchus (nematoda:tylenchida) with descriptions of two species from Michigan. AB - Malenchus truncatus n. sp. and M. bryanti n. sp. from Michigan are describes and presented es the first record of Malenchus in North America. M. acarayensis Andrassy, 1968 is recorded and males of this species described. Notes on the geographical distribution of Malenchus and a key to the species are given. PMID- 19308198 TI - Comparative influence of Radopholus similis and Pratylenchus coffeae on citrus. AB - Pratylenchus coffeae was as pathogenic as Radopholus sirnilis to commercial citrus rootstocks. No rootstock resistant to R. similis was resistant to P. coffeae. Both nematodes stunted citrus in three soil types. Seedling damage by P. coffeae and R. similis was greatest in fine- and coarse-textured soils, respectively. Reproduction and survival on citrus were greater for P. coffeae than for R. similis. Mixed inoculations with R. similis and P. coffeae resulted in lower populations of each species than did separate inoculations. PMID- 19308199 TI - Biology of Anguina plantaginis parasitic on Plantago aristata. AB - Among 17 species and cultivars of plants exposed to Anguina plantaginis, only Plantago aristata (bracted plantain) was a host. Larvae penetrated the emerging apical meristem; reproduces and migrated progressively; caused twisting and galling of leaves, looping andspiraling of peduncles, and transformation of floral structures into galls. Extreme infections caused stunting and death of entire plants. Hypertrophy and hyperplasia of leaf mesophyll, cell separation and disintegration, and xylem wall thickening in older galls occurred. Only third stage larvae were infective, and they exhibited cryptobiosis under adverse conditions. PMID- 19308200 TI - Ultrastructure of the spicules of Pratylenchus penetrans. AB - The two spicules of PratyIenchus penetrans are not morphologically identical. The outer spicule is slightly larger and longer than the inner spicule. Each spicule is composed of a blade, shaft, and base. The spicules originate from the gradual thickening and invagination of the posterior walls of the spicular pouch. At the tip of each spicule are two small pores with one dendritic process associated with each pore. At the shaft portion, the spicule has a thick body which has a central core filled with nerve tissue. Dorsal and ventral wings project from the body. When protruded, the two wings of a given spicule curve toward the corresponding wings of the other spicule to form a complete tube for the conduction of sperm. The base of each spicule is composed of an electron-dense, sclerotized ring with nerve tissue in the central core. Each spicule is mobilized by two protractor and two retractor muscles. The posterior lips (=hypoptygma) of the cloaca are prominent and innervated. The vas deferens and rectum join near the cloacal opening. PMID- 19308201 TI - Interactions of concomitant species of nematodes and Fusarium oxysporum f. sp. vasinfectum on cotton. AB - Meloidogyne incognita, Hoplolaintus galeatus, and North Carolina and Georgia populations of Belonolaimus longicaudatus were introduced singly and in various combinations with Fusarium oxysporum f. sp. vasinfectum on wilt-susceptible 'Rowden' cotton. Of all the nematodes, the combination of the N. C. population of B. longicaudatus with Fusarium promoted greatest wilt development. H. galeatus had no effect on wilt. With Fusarium plus M. incognito or B. longicaudatus, high nematode levels promoted greater wilt than low levels. The combination of either population of B. longicaudatus with M. incognita and Fusarium induced greater wilt development than comparable inoculum densities of either nematode alone or where H. galeatus was substituted for either of these nematodes. Nematode reproduction was inversely related to wilt development. Without Fusarium, however, the high inoculum level resulted in greater reproduction of all nematode species on cotton. Combining M. incognita with B. longicaudatus or H. galeatus gave mutually depressive effects on final nematode populations. The interactions of H. gateatus with B. longicaudatus varied with two populations of the latter. PMID- 19308203 TI - Predicting on-set of egg production by Meloidogyne hapla on lettuce from field soil temperatures. PMID- 19308202 TI - The influence of Trichoderma harzianum on the root-knot Fusarium wilt complex in cotton. AB - Wilt-susceptible cultivar 'Rowden' cotton was inoculated wilh Meloidogyne incognita (N), Trichoderma harzianum (T), and Fusarium oxysporum f. sp. vasinfectum (F) alone and in all combinations in various time sequences. Plants inoculated with F alone or in combination with T did not develop wilt, Simultaneous inoculation of 7-day-old seedlings with all three organisms (NTF) produced earliest wilt. However, plants receiving nematodes at 7 days and Fusarium and Trichoderma at 2 or 4 weeks later (N-T-F, N-TF) developed the greatest wilt between 49-84 days after initial nematode inoculation. During the same period, Fusarium added 4 weeks after initial nematode inoculation (N-F) and Fusarium added 4 weeks after initial simultaneous inoculation of nematode and Trichoderma (NT-F) produced the least wilt. The addition of Fusarium inhibited nematode reproduction. Simultaneous inoculation with nematodes and Trichoderma (NT-) resulted in the greatest root gall development, whereas nematodes alone produced the greatest number of larvae. In comparison with noninoculated controls (CK), treatments involving all three organisms inhibited plant growth, plants inoculated with the nematode alone (N-) or with nematodes and Trichoderma (NT-) simultaneously had greatest root weight. Any treatment involving the nematode resulted in fewer bolls per plant and greater necrosis on roots than the noninoculated checks. PMID- 19308204 TI - An outer layer on eggs of Dolichodorus heterocephalus (Tylenchida). PMID- 19308205 TI - Electrical stimulation applied to second-stage larvae of Heterodera rostochiensis to determine viability. PMID- 19308206 TI - Somatic centrioles in the parasitic nematode, Capillaria hepatica Bancroft, 1893. PMID- 19308207 TI - Revision of the genus Paratylenchus Micoletzky, 1922 and descriptions of new species. Part III of three parts-Gracilacus. AB - Part III covers species with female stylet length >41 microm which are considered by this author to comprise the genus Gracilacus Raski, 1962. Seven new species of Gracilacus are described and further observations given on 14 other species. Paratylenchus strenzkei (Volz, 1951) Oostenbrink, 1960 is transferred to species inquirendae. A key to the species of Gracilacus is included. PMID- 19308208 TI - Effects of potassium fertilization and Pratylenchus penetrans on yield and potassium content of red clover and alfalfa. AB - Red clover and alfalfa were inoculated with Pratylenchus penetrans and grown in an Alberry sandy loam soil to which potassium (K) was added at seeding at 0, 41.5, 83, and 166 microg/g. In one experiment with alfalfa, additional K was added after each forage cut to replace that which was removed. Nematode populations were not consistently affected by K fertilization. Nematode infection stunted red clover and alfalfa and resulted in lower yields at all K levels, except for alfalfa at the lowest K level. Nematode infection had no effect on taproot yields. However, it resulted in lower rootlet yields from red clover at all K levels, lower rootlet yields from alfalfa only at the highest K level in one experiment, and lower rootlet yields at all but the lowest K level in a second experiment, potassium fertilization enhanced yield of red clover and alfalfa. Yield increases were smaller from increased K fertilization in nematode infested soil than in noninfested soil. Pralylenchus penetrans had little effect on the K content of red clover or alfalfa. The stunting of plants from nematode infection resulted in less K being removed from the soil. PMID- 19308209 TI - Effects of oxamyl on the citrus nematode, Tylenchulus semipenetrans, and on infection of sweet orange. AB - Foliar sprays of 4 microg/ml oxamyl on sweet orange trees in a greenhouse slightly depressed the number of Tylenchulus semipenetrans larvae obtained from roots and soil, but similar treatments were not effective in two orchards. Soil drench treatments decreased the number of citrus nematode larvae obtained from roots or soil of citrus plants grown itt a greenhouse and in orchards. Exposure to 5-10 microg/ml of oxamyl in water was lethal to only a few second-stage larvae treated 10 days, and many second-stage larvae in 2.0 microg/ml oxamyl recovered motility when transferred to fresh water. Aqueous solutions of 50 and 100 microg/ml of oxamyl were toxic to citrus nematode larvae. Additional observations indicate that oxamyl interfered with hatch of citrus nematode larvae and was nematistatic and/or protected sweet orange roots from infection. Oxamyl degraded at different rates in two soils. The number of citrus nematode larvae that infected and developed on sweet orange roots was increased by an undetermined product of the degradation of oxamyl in soil, water, and possibly within plants. This product apparently was translocated in roots. PMID- 19308210 TI - Nematode community structure in a vineyard soil. AB - Distribution of the nematode community in a California vineyard was studied over a 13-month period. Omnivorous and microbivorous nematodes were similarly distributed in the root zone, with greatest densities occurring between vine rows and near the soil surface. Greatest densities of plant-parasitic nematodes were found in the vine row, with the individual species differing in their vertical distribution. Total nematode biomass was greatest between rows near the surface. Biomass of plant parasites was greatest in the upper 30 cm of soil in the row, whereas biomass of microbivores was greatest in this region between rows. Of the plant-parasitic nematodes, the variability in distribution among vines was greatest for Paratylenchus hamatus and least for Meloidogyne spp. PMID- 19308211 TI - Effects of oxime carbamate nematicides on development of Heterodera schachtii on sugarbeet. AB - Treatment of sugarbeet, Beta vulgaris L., with aldicarb, aldicarb sulfoxide, or aldicarb sulfone 10 days after plants were inoculated with Heterodera schachtii prevented development of the nematode, but second-stage larvae penetrated the roots. These chemicals had no measurable effects on nematodes in plants treated 15 days after inoculation. The tests established that soil treatments of aldicarb are directly or indirectly lethal to larvae developing within roots of sugarbeet. Heterodera schachtii failed to develop on root slices of red table beet grown in soil treated with aldicarb or aldicarb sulfoxide. Similar treatment of plants with aldicarb sulfone or oxamyl did not affect subsequent development of H. schachtii on root slices of treated plants. PMID- 19308212 TI - Host-parasite relationships of Hoplolaimus columbus on Cotton and soybean. AB - Hoplolaimus columbus suppressed growth and pod yield of soybean in greenhouse tests. Although populations of H. columbus decreased in short-term experiments, increases occurred in long-term studies. The nematode caused extensive damage to the cortical parenchyma and occasionally to the endodermal-vascular region of both cotton and soybean roots. The nematode frequently entered secondary root primordia. Roots of soybean parasitized by H. columbus at high inoculum levels were severely damaged. The relationship of populations of H. columbus and stunting of soybean and cotton is discussed. PMID- 19308213 TI - Vertical migration of the rice white-tip nematode, Aphelenchoides besseyi. AB - In the laboratory, the vertical migration of Aphelenchoides besseyi was favored by rough surfaces and an inverse water gradient. In relation th gravity, the nematode migrated down or up equally, a circumstance suggesting that a geotaxis was not involved. Stems of rice seedlings were effective surfaces for vertical migration of nematodes only when the stems were continuously supplied with moisture. PMID- 19308214 TI - Adenosine triphosphate quantification as related to cryptobiosis, nematode eggs, and larvae. AB - Sonification was the most effective method used for disintegrating nematode eggs and larvae for adenosine triphosphate (ATP) determinations. Sensitivity of the assay was sufficient to measure ATP in one larva. Second-stage larvae of Anguina tritici averaged 1 x 10 femtograms (fg) ATP and Meloidogyne incognita eggs, 0.8 x 105 fg ATP. Larvae of Panagrellus redivivus, a saprobe, averaged 12.2 x 105 fg ATP, a measurement which was considerably higher than the ATP levels in plant parasites. Endophytic bacteria and fungi from wheat galls were detected as background organisms associated with A. tritiei activated by hydration. Also, bacteria in suspensions of eggs from M. incognita prepared with NaCIO were measured by the use of butanol extraction and ATP determination. Second-stage A. tritici larvae increased in ATP content within 40 min after being activated from cryptobiosis by hydration. In the cryptobiotic state, larvae had 50% less ATP than when active. ATP concentrations were similar in galls of different ages. Apparently, ATP concentrations do not change during cryptobiosis. Starvation results in a decline in ATP concentration/larva. Subjecting A. tritici larvae to the lethal temperature of 60 C resulted in a three-fold increase in the decay rate of ATP over that of larvae sonified, then heated at 60 C. These results suggest an association between ATP decay and the mechanism that causes death of larvae at elevated temperatures. PMID- 19308215 TI - Chromonema heliothidis n. gen., n. sp. (Steinernematidae, Nematoda), a parasite of Heliothis zea (Noctuidae, Lepidoptera), and other insects. AB - Chromonema heliothidis n. gen., n. sp. is described as an entomophilic nematode of Heliothis zea and other lepidopterous larvae; the diagnosis of the family Steinernermatidae is emended. In most morphological and host-parasite features, this nematode is similar to neoaplectanid nematodes; however, males are different in having a peloderan bursa and straight to slightly curved spicules. Although the infective-stage juveniles only give rise to hermaphrodites, the nematode is heterogonic, with both males and females being produced in the second generation. Parasitized hosts are brick-red in color and luminescent in the dark because of the association of a chromogenic, bioluminescent bacterium with the nematode. The nematode is capable of parasitizing a wide range of insects with lepidopterous larvae being most susceptible. PMID- 19308217 TI - Extracting the rice white-tip nematode, Aphelenchoides besseyi, with match sticks. PMID- 19308216 TI - Gametogenesis in amphimictic and parthenogenetic populations of Aphelenchus avenae. AB - The female reproductive system of Aphelenchus avenae, studied in orcein-stained material, showed a peculiar structural pattern not yet reported in other nematodes. Chromosome morphology and behavior during gametogenesis could be studied in more detail than in other tylenchid or aphelenchid species investigated to date. In a bisexual population from Australia, gametogenesis was by normal meiosis and reproduction by amphimixis. The haploid chromosome number was n=8 in both males and females, and no sex chromosomes were detected. Three monosexual populations from Australia, California, and North Carolina underwent oogenesis by meiosis but reproduced hy parthenogenesis. The haploid chromosome number was n=8 in the Australia and the North Carolina populations, but n=9 in the California population. Spermatogenesis in temperature-induced males of the California population was by normal meiosis, and sperm had n=9 chromosomes. Most chromosomes consisted of a central euchromatic section and two characteristic heterochromatic ends. No centromere was observed in any chromosome. The relationship between the California population with n=9 and all the other populations with n=8 chromosomes is not well understood. PMID- 19308218 TI - SEM observations on nematode cuticle penetration by Bacillus penetrans. PMID- 19308219 TI - Effects of plastic mulch on soil treatments toxic to Pratylenchus penetrans. PMID- 19308220 TI - Observations on the distribution of Meloidogyne spp. in a vineyard soil. PMID- 19308221 TI - Morphological variation in Pratylenchus penetrans. AB - Variability of morphological characters used to separate Pratylenchus penetrans from other species of the genus was studied in a population originating from a single gravid female. Pronounced heteromorphism was observed and characterized. About 30% of females had a crenate-tail terminus. Several shapes of stylet knobs were characterized; 50% of them were anteriorly flattened to indented. The outer margin of the cephalic framework extended into the body from one-half to two annules. The shape of the spermatheca varied from round to oval. A fifth lateral line was observed in many specimens. Environmental factors, and particularly the host plant, influenced such morphometric characters as body length, width, esophagus length, stylet length, V value, a and b' ratios, as well as qualitative characters such as tail terminus, growth of ovary, and shape of median bulb. Nematodes reared on pea and cabbage had a higher percentage of females with a crenate-tail terminus than those from tomato, rye, beet, and alfalfa callus culture. Nematodes from peas were longer and wider; they often had gonads that extended to esophagi, but they had shorter esophagi amt stylets than those from callus culture. Populations from different geographical locations also exhibited variahility in morphological characters, as did the Cornell population. The validity of many characters used in species identification is discussed, and the possibility that other related Pratylenchus species are conspecific with P. penetrans is suggested. PMID- 19308222 TI - Sex expression and tail morphology of female progenies of smooth-tail and crenate tail females of Pratylenchus penetrans. AB - An analysis of the offspring of single smooth- and crenate-tail females of Pratylenchus penetrans indicated the existence of progenies containing only males or females. Of the 80 progenies analyzed, 46 contained females with smooth and crenate tails. In general, regardless of the mother's tail type, most females possessed crenate tails, although more crenate-tail females originated from a crenate-tail female than from a smooth-tail female. Twenty-three progenies contained only females with crenate tails, most of them originating from crenate tail females. One progeny originating from a smooth-tail female contained only females with smooth tails. No simple interpretation of the inheritance of tail type could be attempted because selection pressure favored females with crenate tails when P. penetrans was reared on Wando pea plants. PMID- 19308223 TI - Parasitism of Beta vulgaris by Meloidogyne hapla and Heterodera schachtii alone and in combination. AB - Interrelationships of Meloidogyne hapla and Heterodera schachtii in combinations of several population levels and different inoculation periods were studied. Results indicated suppression of gall development of M. hapla in any treatment in which inoculations of H. schachtii preceded those of M. hapla by 10 days. This interrelationship was characterized by amensalism with M. hapla serving as an amensat and H. schachtii serving as an inhibitor. Conversely, when inoculations of M. hapla preceded H. schachtii inoculations by 10 days, there were increases in cyst development. This relationship was characterized by commensalism with H. schachtii serving as a commensal. In both interactions, the preinvading parasites acted independently and established populations equal to treatments receiving either parasite alone. When both nematodes were inoculated simultaneously, there were no effects on populations of either. Relationships of this nature were characterized by neutralism. Ratios of total soluble/reducing carbohydrates were lower in treatments when M. hapla inoculations preceded those of H. schachtii. Plants inoculated with both nematodes died earlier than those inoculated with either parasite alone. High concentrations of Al and Fe occurred in treatments wherein M. hapla or H. schachtii inoculations preceded each other by 10 days. Generally, noninoculated control plants exhibited higher concentrations of K, P, Mg, and B than other treatments. PMID- 19308224 TI - Two semi-automatic elutriators for extracting nematodes and certain fungi from soil. AB - Two efficient, semi-automatic elutriators for assaying soil samples for nematodes are described. The first apparatus is a four-unit elutriator which combines conventional extraction methods with the following major features: automatic mixing of 500- to 1,500-cm(3) soil samples with water (+/- air); "turbinate" sample splitters from which fractions of 1/15 or greater are passed onto 26- or 38-microm sieves for collection of larvae and adult nematodes; the capacity for collecting roots, intact egg masses, and cysts on 250-425-microm sieves; and a variable speed motorized sieve-shaker. Nematodes, after being collected on 38 microm sieves, are separated from debris by centrifugation or by Baermann trays. Secondary features include: air cylinders, solenoid valves, and time clock for automatic dumping residual soil and water; relay-controlled coarse spray nozzles activated for 5 sec every 30 sec for washing nematodes through 250-425-mum sieves; adjustable rates of water amt air flow, and tinting. The second type of elutriator operates on similar principles but costs less to construct. It requires somewhat more operator participation; sieve spraying is carried out by the operator, anti elutriators are dumped manually. Both elutriators also show promise for monitoring populations of certain other soil microorganisms. PMID- 19308225 TI - Funaria maryanneae n. sp. and Proleptonchus weischeri n. sp. (Nematoda: Dorylaimida) from Europe and new synonyms in Leptonchoidea. AB - Funaria maryanneae n. sp., distinguished by its large size and long prerectum, is described from specimens collected in Bad Sooden, Germany, D. B. R. This nematode was collected also from Frakmuntegg (Mt. Pilatus), Switzerland. Proleptonchus weischeri n. sp., with short hemispheroid female tail and relatively anterior vulva, is described from specimens collected in Kaufunger Wald near Kassel, Germany, D. B. R. This is the first report of either genus from the continent of Europe. Leptonchus parisii Zullini, 1973, from Chiapas, Mexico, becomes a junior synonym of Funaria obtusa (Thorne, 1939) Goseco, Ferris and Ferris, 1974. Leptonchus acutus Zulliui, 1973 becomes Funaria acuta n. comb., and Funaria rothi Goseco, Ferris and Ferris, 1974 becomes a junior synonym of Funaria acuta. PMID- 19308226 TI - Fine structure of the esophagus of Pratylenchus penetrans. AB - The fine structure of the esophagus of Pratylenchus penetrans is described. The gland lobe is syncytial and contains two types of nuclei: three large nuclei with little chromatin, and more numerous smaller nuclei with large amounts of chromatin. Some of the smaller nuclei are associated only with glandular tissue, whereas others are part of nerve ceils within the esophagus. Clusters of free ribosomes, rough endoplasmic reticulum, and numerous mitochondria occur in the lobe region where the secretory granules are formed. No Golgi bodies were observed. On the basis of these observations, possible differences in the mechanism of secretory granule formation between plant-parasitic nematodes are discussed. Several other minor differences between the fine structure of other plant-parasitic nematodes previously examined and that of P. penetrans are also noted. PMID- 19308227 TI - Life cycle, pathogenicity, histopathology, and host range of race 5 of the barley root-knot nematode. AB - The optimum temperature for development of race 5 of Meloidogyne naasi was 26 C. A life cycle was completed in 34 days. Growth of sorghum was suppressed when inoculated with M. naasi. Observations of M. naasi-infected sorghum roots demonstrated that roots were penetrated just behind the root cap; giant cells were generally initiated either in the procambial region or in very young phloem. When giant cells developed in the cortex, corresponding areas of the vascular system did not have an endodermis, pericycle, or phloem fibers. Nineteen plant species were tested for suitability as hosts for race 5 of M. naasi. Reproduction occurred on 11 of 12 monocotolydenous hosts and none of 7 dicotolydenous hosts. Reproduction often occurred without gall development. PMID- 19308228 TI - Relationships of initial population densities of Meloidogyne incognita and M. hapla to yield of tomato. AB - Microplots 80 x 100 cm, infested with varying initial population densities (P(i)) of Meloidogyne incognita or M. hapla, were planted to tomato at two locations. Experiments were conducted in a sandy loam soil at Fletcher, N. C. (mountains) where the mean temperature for May to September is ca 20.7 C, and in a loamy saml at Clayton, N. C. (coastal plain) where the mean temperature for May to Septemher is ca 24.8 C. In these experimentally infested plots, M. incognita and M. hapla caused maximum yield losses of 20-30%, at lhe mountain site with Pi of 0-12,500 eggs and larvae/500 cm(3) of soil. In the coastal plain, M. incognita suppressed yields up to 85%, and M. hapla suppressed yields up to 50% in comparison with the non infested control. A part of the high losses at this site apparently was due to M. incognita predisposing tomato to the early blight fungus. In a second experiment, in which a nematicide was used to obtain a range of P(i)s (with P(i) as high as 25,000/50 cm(3) of soil) at Fletcher, losses due to M. incognita were as great as 50%, but similar densities of M. hapla suppressed yields by only 10 25%. Approximate threshold densities for both species ranged from 500 to 1,000 larvae and eggs (higher for surviving larvae) for the mountain site, whereas numbers as low as 20 larvae/500 cm(3) of soil of either species caused significant damage in the coastal plain. Chemical soil treatments proved useful in obtaining various initial population densities; however, problems were encountered in measuring effective inoculum after such treatments, especially in the heavier soil. PMID- 19308229 TI - Relative susceptibility of selected cultivars, of potato to Pratylenchus penetrans. AB - Pratylenchus penetrans suppressed the tuber yields of potato cultivars 'Katahdin', 'Kennebec', and 'Superior', but did not affect yields of 'Russet Burbank'. In comparison with noninfested controls, all initial nematode densities (P(i)) of P. penetrans (P(i) = 38, 81, 164, 211/ 100 cm(3) of soil) suppressed yields of Superior; a moderate P(i) (81/100 cm(3) soil) suppressed yields of Kennebec; and on Katahdin, a moderate P(i) enhanced yields, but higher P(i)'s caused a marked loss. In general, yields were related to the tolerance of the cultivars to nematode colonization. Highest nematode densities were found in the roots of Russet Burbank; the next highest, in succeeding order, were found in roots of Kennebec, Katahdin, and Superior. Symptoms of nematode invasion were confined to losses of tuber yield and root weight. PMID- 19308230 TI - Vertical distribution of three namatode species in relation to certain soil properties. AB - Population densities of Belonolaimus longicaudatus, Pratylenchus brachyurus, and Trichodorus christiei were determined from soil samples taken weekly in Tifton, Georgia during a 14-month period (except for April and May) at 15-cm increments to a depth of 105 cm. Belonolaimus longicaudatus predominately inhabited the top 30 cm of soil that was 87-88% sand, 6-7% silt, and 5-7% clay. No specimens were found below 60 cm where the soil was 76-79% sand, 5-6% silt, and 15-19% clay. Highest population densities occurred during June through September when temperature in the top 30 cm of soil was 22-25 C and soil moisture was from 9 to 20% by volume. Pratylenchus brachyurus was found at all depths, but population densities were greatest 45-75 cm deep where the soil was 78-79% sand, 6% silt, and 15-16% clay. In the months monitored, highest population densities occurred during March, June, and December when the soil temperature 45-75 cm deep was 14 17 C and soil moisture was 22-42%. Trichodorus christiei was found at all depths, but population densities were highest 30 cm deep where the soil was 83% sand, 5% silt, and 12% clay. Highest population densities occurred during December through March when the soil temperature 30 cm deep was 11-17 C and soil moisture was 18 23%. PMID- 19308231 TI - Observations on development of the gonad and on reproduction in Aphelenchus avenae. AB - Nuclear changes occurring in male and female gonads of Aphelenchus avenae during postembryogenesis were studied in relation to time and feeding periods on Rhizoetonia solani. Development of the female gonad was similar to that in other nematode species, but development of the male gonad followed the growth pattern of female rather than male gonads. This deviation was explained by the assumption that males in antphimictic populations have appeared as the resnlt of recent evolntion of such populations to sexuality from originally parthenogenctic ancestors. A certain, period of feeding of larvae (16 h for L-2 and L-3, but 12 h for L-4) was required before molting. Cell divisions were confined to the periods of lethargus during the second and third molts, but started during the larval stage in fourth-stage larvae. Crosses in various combinations demonstrated that temperature-induced males do inseminate fentales of the amphimictic and some parthenogenetic populations, but their spermatozoa are nonfunctional. Similarly, males of the amphimictic population inseminated females of a parthenogenetic population, but the sperm did not penetrate the oocytes. PMID- 19308232 TI - Development of a computer-simulation model for a plant-nematode system. AB - A computer-simulation model (MELSIM) of a Meloidogyne-grapevine system is developed. The objective is to attempt a holistic approach to the study of nematode population dynamics by using experimental data from controlled environmental conditions. A simulator with predictive ability would be useful in considering pest management alternatives and in teaching. Rates of flow and interaction between the components of the system are governed by environmental conditions. Equations for these rates are determined by fitting curves to data from controlled environment studies. Development of the model and trial simulations have revealed deficiencies in understanding of the system and identified areas where further research is necessary. PMID- 19308234 TI - Anesthetization of morphologically similar species with carbon dioxide for separation and culture. PMID- 19308233 TI - Host plants, distribution, and ecological association of Hoplolaimus columbus. AB - Selected species and cultivars of plants were evaluated for host suitability for Hoplolaimus columbus under greenhouse and fieht conditions. Sixteen agronomic plant cultivars were assayed for infection and reproduction after 2-6 months. Lima bean, soybean, cotton, and sweet corn were most favorable for nematode reproduction, whereas sweet potato was a nonhost plant. Field corn and watermelon, which tolerated H. columbus without significant yield losses, are suited for alternative crops in the southern coastal plain. Populations of H. columbus occurred in 15% of soyhean and 25% of cotton soil samples assayed. Population levels of H. columbus and Helicotylenchus spp. were correlated with fluctuations in various soil nutrient factors, whereas Meloidogyne spp. and Scutellonema spp. were not. No correlation was detected between the presence of H. columbus and populations of other nematode genera stndied. PMID- 19308235 TI - Scanning electron microscopic observations of Duboscqia penetrans parasitizing root-knot larvae. PMID- 19308236 TI - Comparative biology of the Wyoming and Louisiana populations of Reesimermis nielseni, parasitic nematode of mosquitoes. PMID- 19308237 TI - The effects of ammonia-generating fertilizer on Criconemoides xenoplax in pot cultures. AB - Fertilizer which generated NH was detrimental to Criconemoides xenoplax grown in the greenhouse. The fertilizer was lethal to the nematodes in vitro only when it was accompanied by urease-positive bacteria or partially purified urease. The detrimental action of the fertilizer-urease mixture was ntore rapitt at pH 8 than at pH 7. PMID- 19308238 TI - Influence of some environmental factors on populations of Pratylenchus minyus in wheat. AB - The distribution and density of Pratylenchus minyus and possible relationships of several environmental components, including ammonium nitrate, were investigated in a wheat field in South Australia. Seasonal variation as measured every 2-4 weeks was eliminated from the observations by periodic regression. Correlation and regression analyses were then used to investigate the association of host plant, rainfall, temperature, and the fungus Gaeumannomyces graminis with P. minyus. Other than seasonal effects, soil moisture and G. graminis were the only components associated with P. minyus. Ammonium nitrate usually was correlated with fewer P. minyus in wheat roots. Much higher numbers of P. minyus were observed in seminal than in crown roots of wheat. PMID- 19308239 TI - Effects of Pratylenchus coffeae and Scutellonema bradys alone and in combination on guinea yam (Dioscorea rotundata). AB - When Guinea yam, Dioscorea rotundata Poir, was inoculated with Pratylenchus coffeae and Scutellonema bradys together, there was a 53% suppression of top growth, but when plants were inoculated separately there was a 29% suppression with P. coffeae and a 21% suppression with S. bradys. The reproduction of S. bradys was greatly inhibited when both nematode species were together on the same plant in comparison with that on plants inoculated with S. bradys alone. Scutellonema bradys apparently did not affect the reproduction of P. coffeae. Storage-root quality was reduced 72% by P. coffeae but only 20% by S. bradys. The two species together resulted in a reduction in storage-root quality of 84%. Although the Guinea yam is a good host of P. coffeae and S. bradys, our results indicate that P. coffeae is responsible for most of the storage-root deterioration and dry rot of yam in Puerto Rico. PMID- 19308240 TI - Effects of birds ingesting Heterodera rostochiensis cysts on viability of eggs and larvae. AB - Rate of passage through the digestive systems and effects of ingestion on viability of contents of cysts of Heterodera rostochiensis were determined in feeding trials with pigeons, thrushes, starlings, cowbirds, sparrows, and quails. Depending upon species of birds, 12-82% of the cysts ingested passed through the digestive system within 0.5 h. Pigeons required 6 h for complete evacuation. All other birds completely evacuated ingested cysts from their digestive systems within 3 h. Contents of cysts were nonviable if they were retained in the digestive system of starlings for more than 1.5 h, pigeons more than 1 h, or other birds more than 0.5 h. Cyst contents were nonviable if they remained in contact with excreta from cowbirds or quails for 4 h, thrushes for 96 h, or other species for 72 h after passage. Viability of contents of cysts was inversely related to exposure to excreta-filtrate concentration. Larvae failed to emerge from cysts that were exposed to a 25% concentration of excreta filtrate from starlings, 50% concentration from pigeons or thrushes, or 100% concentration of excreta filtrates from each of the other species. Cysts that were subjected to 44 C (avg. body temperature of cowbirds) for more than 3.5 h were nonviable. Cysts that passed through birds and collected with excrement on polyethylene or soil produced no infective larvae on potato. PMID- 19308241 TI - Effects of density of Helicotylenchus dihystera and Pratylenchus vulnus on American boxwood growing in microplots. AB - American boxwood, Buxus sempervirens var. globosum, was tolerant of Helicotylenchus dihystera [in field microplots] as measures of plant growth were similar to the control and nematode densities were maintained at high levels (1,705-1,810/500 cm(3) soil after 29 months). Boxwood was intolerant of Pratylenchus vulnus at initial densities of 163, 281, or 475 nematodes per 500 cm(3) soil. In comparisons with those of controls, vigor ratings of boxwood after 14 months were much lower at all densities of this nematode. Nematode density was not directly related to vigor rating. However, initial nematode density was directly proportional to growth suppression of boxwood as measured by the difference of the product of final plant height x width or by the difference of the plant surface area determined from a standardized photograph as compared to those of controls. A nematode density of 160/500 cm(3) of soil was found to suppress growth by 50%. Populations of P. vulnus declined, according to a linear function, with time after reaching over 7,100 nematodes/500 cm(3) of soil taken from the root zones of boxwoods. Ninety-five percent of the P. vulnus population died between 15 and 20 months after soil infestation. PMID- 19308242 TI - Discocriconemella repleta n.sp., and the male of Criconemoides inusitatus Hoffmann, 1974 (Criconematidae: Nematoda). AB - Discocriconemella repleta n.sp. from Brazil is described. The male of Criconemoides inusitatus Hoffmann, 1974, is described here for the first time. PMID- 19308243 TI - Effects of Pratylenchus vulnus and Xiphinema index singly and combined on vine growth of Vitis vinifera. AB - Inoculation of 'Thompson Seedless' grapevines with 500 Xiphinerna index or 1,000 Pratylenchus vulnus alone or in combination suppressed vine shoot and root growth under greenhouse conditions. Pratytenchus vulnus caused greater stunting of roots than X. index. Each nematode species inhibited top growth about equally. Concomitant inoculations caused greater stunting of tops and roots than did inoculations of either nematode species alone. Differences in growth between inoculated and control plants increased with exposure time. Pratylenchus vulnus competed with and gradually superseded in numbers an established population of X. index. Both species reproduced on 'Thompson Seedless' roots, but P. vulnus increased to a much higher level than did X. index. The increase of P. vulnus, together with extensive damage, proves its pathogenicity to grapevines. PMID- 19308244 TI - Histopathology of Beta vulgaris to individual and concomitant infections by Meloidogyne hapla and Heterodera schachtii. AB - Histological changes in roots of Beta vulgaris cultivar 'USH9A' resulting from infection of Meloidogyne hapla alone, Heterodera schachtii alone, or infection by both species on one feeding site were studied. Anatomical changes caused by M. hapla infection were characterized by regions of hypertrophy and hyperplasia. Giant cells were formed within the stele and varied in numbers from 4-7/feeding site; hyperplasia occurred in the form of a large number of relatively small compacted cells generally surrounding the hypertrophied region. H. schachtii induced syncytia became dense and muhinucleate. Syncytia were formed in the stele and were limited on the side toward the nematode by endodermis or in part by cortical tissue. Histological changes due to the presence of both parasites on one feeding site were characterized by formation of two distinctive pathological tissues typical of both nematodes. In most infections, xylem elements separated the two pathological tissues. In some sections, a single wall separated the two pathological tissues, and no dissolution of separating wall was noted in any sections. Each nematode developed normally and produced its own characteristic pathological tissue independently. PMID- 19308245 TI - Efficacy of multiple applications of oxamyl and phenamiphos for control of Pratylenchus penetrans in birdsfoot trefoil. AB - Preplant soil applications of granular phenamiphos effectively reduced Pratytenchus penetrans in soil during the seeding year and 1 year after, and in the roots of birdsfoot trefoil 2 years after seeding. Forage yields were increased in the season following application of phenamiphos, but stands of plants/m(2) were not greater (P = 0.05) than those in the checks 1 and 2 years after treatment. Additional spring applications of phenamiphos 1 and 2 years after seeding further reduced numbers of nematodes in the soil but did not improve forage yields or plant stand over that of a single application. Broadcast preplant soil sprays of oxamyl followed by several foliar sprays at different rates and frequencies of application over a 3-year period restricted populations of P. penetrans in the soil and roots of birdsfoot trefoil but did not consistently result in increased forage yields. Stands of birdsfoot trefoil continued to decline each year even with oxamyl treatments. PMID- 19308246 TI - Pathology and histopathology of Pratylenchus scribneri infecting snap bean and lima bean. AB - The pathological effects of Pratylenchus scribneri on susceptible snap bean and resistant lima bean were studied. In a pathogenicity test, the nematode increased nearly 75-fold on snap bean and suppressed top growth. On lima bean, P. scribneri reproduced slowly and did not significantly affect top growth. Discreet lesions formed on lima bean roots, but no lesions developed on snap bean roots. Paraffin sections taken 2, 5, 13, 25, and 32 days after inoculation showed little cellular necrosis in snap bean roots, whereas cells surrounding the nematode in lima bean were extensively necrotic. Cortical cells of infected snap bean roots were almost completely invaded and killed 25 and 32 days after inoculation. The cortex of lima bean tissues was intact, although localized necrotic areas remained at sites of nematode invasion. PMID- 19308248 TI - Vertical Distribution of Hoplolaimus galeatus and oligochaetes in greenhouse colonies. PMID- 19308247 TI - Chemotactic responses of male Caenorhabditis elegans. AB - Cultures of C. elegans containing a high proportion of males were subjected to chemotactic tests by using the method of countercurrent separation. The responses of males and hermaphrodites were determined. Both types of worms preferred Na over 1/2 Ca(2), Cl over NO; they were attracted to NaCl, OH, cyclic AMP, pyridine, CO, in borate buffer (pH 8.8); and avoided CO in phosphate buffer (pH 6.0), D-tryptophan, and acid. It was thus concluded that male C. elegans have the same chemotactic responses that hermaphrodites of this species are known to have. PMID- 19308249 TI - Chamber for critical-point drying of nematodes and other biological specimens. PMID- 19308250 TI - Cathepsin F cysteine protease of the human liver fluke, Opisthorchis viverrini. AB - BACKGROUND: The liver fluke Opisthorchis viverrini is classified as a class I carcinogen due to the association between cholangiocarcinoma and chronic O. viverrini infection. During its feeding activity within the bile duct, the parasite secretes several cathepsin F cysteine proteases that may induce or contribute to the pathologies associated with hepatobiliary abnormalities. METHODOLOGY/PRINCIPAL FINDINGS: Here, we describe the cDNA, gene organization, phylogenetic relationships, immunolocalization, and functional characterization of the cathepsin F cysteine protease gene, here termed Ov-cf-1, from O. viverrini. The full length mRNA of 1020 nucleotides (nt) encoded a 326 amino acid zymogen consisting of a predicted signal peptide (18 amino acids, aa), prosegment (95 aa), and mature protease (213 aa). BLAST analysis using the Ov-CF-1 protein as the query revealed that the protease shared identity with cathepsin F-like cysteine proteases of other trematodes, including Clonorchis sinensis (81%), Paragonimus westermani (58%), Schistosoma mansoni and S. japonicum (52%), and with vertebrate cathepsin F (51%). Transcripts encoding the protease were detected in all developmental stages that parasitize the mammalian host. The Ov cf-1 gene, of approximately 3 kb in length, included seven exons interrupted by six introns; the exons ranged from 69 to 267 bp in length, the introns from 43 to 1,060 bp. The six intron/exon boundaries of Ov-cf-1 were conserved with intron/exon boundaries in the human cathepsin F gene, although the gene structure of human cathepsin F is more complex. Unlike Ov-CF-1, human cathepsin F zymogen includes a cystatin domain in the prosegment region. Phylogenetic analysis revealed that the fluke, human, and other cathepsin Fs branched together in a clade discrete from the cathepsin L cysteine proteases. A recombinant Ov-CF-1 zymogen that displayed low-level activity was expressed in the yeast Pichia pastoris. Although the recombinant protease did not autocatalytically process and activate to a mature enzyme, trans-processing by Fasciola hepatica cathepsin L cleaved the prosegment of Ov-CF-1, releasing a mature cathepsin F with activity against the peptide Z-Phe-Arg-NHMec >50 times that of the zymogen. Immunocytochemistry using antibodies raised against the recombinant enzyme showed that Ov-CF-1 is expressed in the gut of the mature hermaphroditic fluke and also in the reproductive structures, including vitelline glands, egg, and testis. Ov CF-1 was detected in bile duct epithelial cells surrounding the flukes several weeks after infection of hamsters with O. viverrini and, in addition, had accumulated in the secondary (small) bile ducts where flukes cannot reach due to their large size. CONCLUSIONS/SIGNIFICANCE: A cathepsin F cysteine protease of the human liver fluke O. viverrini has been characterized at the gene and protein level. Secretion of this protease may contribute to the hepatobiliary abnormalities, including cholangiocarcinogenesis, observed in individuals infected with this parasite. PMID- 19308251 TI - Assays to detect beta-tubulin codon 200 polymorphism in Trichuris trichiura and Ascaris lumbricoides. AB - BACKGROUND: The soil-transmitted helminths (STH) Ascaris lumbricoides and Trichuris trichiura are gastrointestinal parasites causing many disabilities to humans, particularly children. The benzimidazole (BZ) drugs, albendazole (ALB) and mebendazole (MBZ), are commonly used for mass treatment for STH. Unfortunately, there is concern that increased use of anthelmintics could select for resistant populations of these human parasites. In veterinary parasites, and lately in filarial nematodes, a single amino acid substitution from phenylalanine to tyrosine, known to be associated with benzimidazole resistance, has been found in parasite beta-tubulin at position 200. We have developed pyrosequencer assays for codon 200 (TTC or TAC) in A. lumbricoides and T. trichiura to screen for this single nucleotide polymorphism (SNP). METHOD AND FINDINGS: Pyrosequencing assays were developed and evaluated for detecting the TTC or TAC SNP at codon 200 in beta-tubulin in A. lumbricoides and T. trichiura. Genomic DNA from individual worms, eggs isolated from individual adult worms or from fecal samples with known treatment history and origin, were sequenced at beta-tubulin by pyrosequencing, and genotypes were confirmed by conventional sequencing. The assays were applied to adult worms from a benzimidazole-naive population in Kenya. Following this, these assays were applied to individual worms and pooled eggs from people in East Africa (Uganda and Zanzibar) and Central America (Panama) where mass anthelmintic drug programs had been implemented. All A. lumbricoides samples were TTC. However, we found 0.4% homozygous TAC/TAC in T. trichiura worms from non-treated people in Kenya, and 63% of T. trichiura egg pools from treated people in Panama contained only TAC. CONCLUSION: Although the codon 200 TAC SNP was not found in any of the A. lumbricoides samples analyzed, a rapid genotyping assay has been developed that can be used to examine larger populations of this parasite and to monitor for possible benzimidazole resistance development. The TAC SNP at codon 200, associated with benzimidazole resistance in other nematodes, does occur in T. trichiura, and a rapid assay has been developed to allow populations of this parasite to be monitored for the frequency of this SNP. Sample sizes were small, anthelmintic efficacy was not assessed, and treated and non-treated samples were from different locations, so these frequencies cannot be extrapolated to other populations of T. trichiura or to a conclusion about resistance to treatment. The occurrence of the TAC SNP at codon 200 of beta-tubulin in T. trichiura may explain why benzimidazole anthelmintics are not always highly effective against this species of STH. These assays will be useful in assessing appropriate treatment in areas of high T. trichiura prevalence and in monitoring for possible resistance development in these STH. PMID- 19308252 TI - Genetic variation in VEGF does not contribute significantly to the risk of congenital cardiovascular malformation. AB - Several previous studies have investigated the role of common promoter variants in the vascular endothelial growth factor (VEGF) gene in causing congenital cardiovascular malformation (CVM). However, results have been discrepant between studies and no study to date has comprehensively characterised variation throughout the gene. We genotyped 771 CVM cases, of whom 595 had the outflow tract malformation Tetralogy of Fallot (TOF), and carried out TDT and case control analyses using haplotype-tagging SNPs in VEGF. We carried out a meta analysis of previous case-control or family-based studies that had typed VEGF promoter SNPs, which included an additional 570 CVM cases. To identify rare variants potentially causative of CVM, we carried out mutation screening in all VEGF exons and splice sites in 93 TOF cases. There was no significant effect of any VEGF haplotype-tagging SNP on the risk of CVM in our analyses of 771 probands. When the results of this and all previous studies were combined, there was no significant effect of the VEGF promoter SNPs rs699947 (OR 1.05 [95% CI 0.95-1.17]); rs1570360 (OR 1.17 [95% CI 0.99-1.26]); and rs2010963 (OR 1.04 [95% CI 0.93-1.16]) on the risk of CVM in 1341 cases. Mutation screening of 93 TOF cases revealed no VEGF coding sequence variants and no changes at splice consensus sequences. Genetic variation in VEGF appears to play a small role, if any, in outflow tract CVM susceptibility. PMID- 19308253 TI - High-throughput proteomics detection of novel splice isoforms in human platelets. AB - Alternative splicing (AS) is an intrinsic regulatory mechanism of all metazoans. Recent findings suggest that 100% of multiexonic human genes give rise to splice isoforms. AS can be specific to tissue type, environment or developmentally regulated. Splice variants have also been implicated in various diseases including cancer. Detection of these variants will enhance our understanding of the complexity of the human genome and provide disease-specific and prognostic biomarkers. We adopted a proteomics approach to identify exon skip events - the most common form of AS. We constructed a database harboring the peptide sequences derived from all hypothetical exon skip junctions in the human genome. Searching tandem mass spectrometry (MS/MS) data against the database allows the detection of exon skip events, directly at the protein level. Here we describe the application of this approach to human platelets, including the mRNA-based verification of novel splice isoforms of ITGA2, NPEPPS and FH. This methodology is applicable to all new or existing MS/MS datasets. PMID- 19308254 TI - Small RNA deep sequencing reveals role for Arabidopsis thaliana RNA-dependent RNA polymerases in viral siRNA biogenesis. AB - RNA silencing functions as an important antiviral defense mechanism in a broad range of eukaryotes. In plants, biogenesis of several classes of endogenous small interfering RNAs (siRNAs) requires RNA-dependent RNA Polymerase (RDR) activities. Members of the RDR family proteins, including RDR1and RDR6, have also been implicated in antiviral defense, although a direct role for RDRs in viral siRNA biogenesis has yet to be demonstrated. Using a crucifer-infecting strain of Tobacco Mosaic Virus (TMV-Cg) and Arabidopsis thaliana as a model system, we analyzed the viral small RNA profile in wild-type plants as well as rdr mutants by applying small RNA deep sequencing technology. Over 100,000 TMV-Cg-specific small RNA reads, mostly of 21- (78.4%) and 22-nucleotide (12.9%) in size and originating predominately (79.9%) from the genomic sense RNA strand, were captured at an early infection stage, yielding the first high-resolution small RNA map for a plant virus. The TMV-Cg genome harbored multiple, highly reproducible small RNA-generating hot spots that corresponded to regions with no apparent local hairpin-forming capacity. Significantly, both the rdr1 and rdr6 mutants exhibited globally reduced levels of viral small RNA production as well as reduced strand bias in viral small RNA population, revealing an important role for these host RDRs in viral siRNA biogenesis. In addition, an informatics analysis showed that a large set of host genes could be potentially targeted by TMV-Cg-derived siRNAs for posttranscriptional silencing. Two of such predicted host targets, which encode a cleavage and polyadenylation specificity factor (CPSF30) and an unknown protein similar to translocon-associated protein alpha (TRAP alpha), respectively, yielded a positive result in cleavage validation by 5'RACE assays. Our data raised the interesting possibility for viral siRNA mediated virus-host interactions that may contribute to viral pathogenicity and host specificity. PMID- 19308255 TI - Borrelia recurrentis employs a novel multifunctional surface protein with anti complement, anti-opsonic and invasive potential to escape innate immunity. AB - Borrelia recurrentis, the etiologic agent of louse-borne relapsing fever in humans, has evolved strategies, including antigenic variation, to evade immune defence, thereby causing severe diseases with high mortality rates. Here we identify for the first time a multifunctional surface lipoprotein of B. recurrentis, termed HcpA, and demonstrate that it binds human complement regulators, Factor H, CFHR-1, and simultaneously, the host protease plasminogen. Cell surface bound factor H was found to retain its activity and to confer resistance to complement attack. Moreover, ectopic expression of HcpA in a B. burgdorferi B313 strain, deficient in Factor H binding proteins, protected the transformed spirochetes from complement-mediated killing. Furthermore, HcpA-bound plasminogen/plasmin endows B. recurrentis with the potential to resist opsonization and to degrade extracellular matrix components. Together, the present study underscores the high virulence potential of B. recurrentis. The elucidation of the molecular basis underlying the versatile strategies of B. recurrentis to escape innate immunity and to persist in human tissues, including the brain, may help to understand the pathological processes underlying louse borne relapsing fever. PMID- 19308256 TI - Maternal protein restriction affects postnatal growth and the expression of key proteins involved in lifespan regulation in mice. AB - We previously reported that maternal protein restriction in rodents influenced the rate of growth in early life and ultimately affected longevity. Low birth weight caused by maternal protein restriction followed by catch-up growth (recuperated animals) was associated with shortened lifespan whereas protein restriction and slow growth during lactation (postnatal low protein: PLP animals) increased lifespan. We aim to explore the mechanistic basis by which these differences arise. Here we investigated effects of maternal diet on organ growth, metabolic parameters and the expression of insulin/IGF1 signalling proteins and Sirt1 in muscle of male mice at weaning. PLP mice which experienced protein restriction during lactation had lower fasting glucose (P = 0.038) and insulin levels (P = 0.046) suggesting improved insulin sensitivity. PLP mice had higher relative weights (adjusted by body weight) of brain (P = 0.0002) and thymus (P = 0.031) compared to controls suggesting that enhanced functional capacity of these two tissues is beneficial to longevity. They also had increased expression of insulin receptor substrate 1 (P = 0.021) and protein kinase C zeta (P = 0.046). Recuperated animals expressed decreased levels of many insulin signalling proteins including PI3 kinase subunits p85alpha (P = 0.018), p110beta (P = 0.048) and protein kinase C zeta (P = 0.006) which may predispose these animals to insulin resistance. Sirt1 protein expression was reduced in recuperated offspring. These observations suggest that maternal protein restriction can affect major metabolic pathways implicated in regulation of lifespan at a young age which may explain the impact of maternal diet on longevity. PMID- 19308257 TI - Cycle inhibiting factors (CIFs) are a growing family of functional cyclomodulins present in invertebrate and mammal bacterial pathogens. AB - The cycle inhibiting factor (Cif) produced by enteropathogenic and enterohemorrhagic Escherichia coli was the first cyclomodulin to be identified that is injected into host cells via the type III secretion machinery. Cif provokes cytopathic effects characterized by G(1) and G(2) cell cycle arrests, accumulation of the cyclin-dependent kinase inhibitors (CKIs) p21(waf1/cip1) and p27(kip1) and formation of actin stress fibres. The X-ray crystal structure of Cif revealed it to be a divergent member of a superfamily of enzymes including cysteine proteases and acetyltransferases that share a conserved catalytic triad. Here we report the discovery and characterization of four Cif homologs encoded by different pathogenic or symbiotic bacteria isolated from vertebrates or invertebrates. Cif homologs from the enterobacteria Yersinia pseudotuberculosis, Photorhabdus luminescens, Photorhabdus asymbiotica and the beta-proteobacterium Burkholderia pseudomallei all induce cytopathic effects identical to those observed with Cif from pathogenic E. coli. Although these Cif homologs are remarkably divergent in primary sequence, the catalytic triad is strictly conserved and was shown to be crucial for cell cycle arrest, cytoskeleton reorganization and CKIs accumulation. These results reveal that Cif proteins form a growing family of cyclomodulins in bacteria that interact with very distinct hosts including insects, nematodes and humans. PMID- 19308258 TI - Discovery and characterization of novel vascular and hematopoietic genes downstream of etsrp in zebrafish. AB - The transcription factor Etsrp is required for vasculogenesis and primitive myelopoiesis in zebrafish. When ectopically expressed, etsrp is sufficient to induce the expression of many vascular and myeloid genes in zebrafish. The mammalian homolog of etsrp, ER71/Etv2, is also essential for vascular and hematopoietic development. To identify genes downstream of etsrp, gain-of function experiments were performed for etsrp in zebrafish embryos followed by transcription profile analysis by microarray. Subsequent in vivo expression studies resulted in the identification of fourteen genes with blood and/or vascular expression, six of these being completely novel. Regulation of these genes by etsrp was confirmed by ectopic induction in etsrp overexpressing embryos and decreased expression in etsrp deficient embryos. Additional functional analysis of two newly discovered genes, hapln1b and sh3gl3, demonstrates their importance in embryonic vascular development. The results described here identify a group of genes downstream of etsrp likely to be critical for vascular and/or myeloid development. PMID- 19308259 TI - Necator americanus infection: a possible cause of altered dendritic cell differentiation and eosinophil profile in chronically infected individuals. AB - BACKGROUND: Hookworms survive for several years (5 to 7 years) in the host lumen, inducing a robust but largely ineffective immune response. Among the most striking aspects of the immune response to hookworm (as with many other helminths) is the ablation of parasite-specific T cell proliferative response (hyporesponsiveness). While the role of the adaptive immune response in human helminth infection has been well investigated, the role of the innate immune responses (e.g., dendritic cells and eosinophils) has received less attention and remains to be clearly elucidated. METHODOLOGY/PRINCIPAL FINDINGS: We report on the differentiation/maturation of host dendritic cells in vitro and the eosinophil activation/function associated with human hookworm infection. Mature DCs (mDCs) from Necator americanus (Necator)-infected individuals showed an impaired differentiation process compared to the mDCs of non-infected individuals, as evidenced by the differential expression of CD11c and CD14. These same hookworm-infected individuals also presented significantly down-regulated expression of CD86, CD1a, HLA-ABC, and HLA-DR. The lower expression of co stimulatory and antigen presentation molecules by hookworm-infected-derived mDCs was further evidenced by their reduced ability to induce cell proliferation. We also showed that this alternative DC differentiation is partially induced by excreted-secreted hookworm products. Conversely, eosinophils from the same individuals showed a highly activated status, with an upregulation of major cell surface markers. Antigen-pulsed eosinophils from N. americanus-infected individuals induced significant cell proliferation of autologous PBMCs, when compared to non-infected individuals. CONCLUSION: Chronic N. americanus infection alters the host's innate immune response, resulting in a possible modulation of the maturation process of DCs, a functional change that may diminish their ability for antigen presentation and thus contribute to the ablation of the parasite-specific T cell proliferative response. Interestingly, a concomitant upregulation of the major cell surface markers of eosinophils was observed in hookworm-infected individuals, indicative of antigen-specific immune responses, especially antigen presentation. We showed that in addition to the postulated role of the eosinophils as effector cells against helminth infection, activated cells may also be recruited to sites of inflammation and contribute to the immune response acting as antigen presenting cells. PMID- 19308260 TI - Detecting linkage between a trait and a marker in a random mating population without pedigree record. AB - Modern linkage-based approaches employing extended pedigrees are becoming powerful tools for localizing complex quantitative trait loci. For these linkage mapping methods, it is necessary to reconstruct extended pedigrees which include living individuals, using extensive pedigree records. Unfortunately, such records are not always easy to obtain and application of the linkage-based approaches has been restricted. Within a finite population under random mating, latent inbreeding rather than non-random inbreeding by consanguineous marriages is expected to occur and is attributable to coalescence in a finite population. Interestingly, it has been revealed that significant random inbreeding exists even in general human populations. Random inbreeding should be used to detect the hidden coancestry between individuals for a particular chromosomal position and it could also have application in linkage mapping methods. Here we present a novel method, named finite population based linkage mapping (FPL) method, to detect linkage between a quantitative trait and a marker via random inbreeding in a finite population without pedigree records. We show how to estimate coancestry for a chromosomal position between individuals by using multipoint Bayesian estimation. Subsequently, we describe the FPL method for detecting linkage via interval mapping method using a nonparametric test. We show that the FPL method does work via simulated data. For a random sample from a finite population, the FPL method is more powerful than a standard pedigree-based linkage mapping method with using genotypes of all parents of the sample. In addition, the FPL method was demonstrated by actual microsatellite genotype data of 750 Japanese individuals that are unrelated according to pedigree records to map a known Psoriasis susceptible locus. For samples without pedigree records, it was suggested that the FPL method require limited number of individuals, therefore would be better than other methods using thousands of individuals. PMID- 19308261 TI - Expert financial advice neurobiologically "Offloads" financial decision-making under risk. AB - BACKGROUND: Financial advice from experts is commonly sought during times of uncertainty. While the field of neuroeconomics has made considerable progress in understanding the neurobiological basis of risky decision-making, the neural mechanisms through which external information, such as advice, is integrated during decision-making are poorly understood. In the current experiment, we investigated the neurobiological basis of the influence of expert advice on financial decisions under risk. METHODOLOGY/PRINCIPAL FINDINGS: While undergoing fMRI scanning, participants made a series of financial choices between a certain payment and a lottery. Choices were made in two conditions: 1) advice from a financial expert about which choice to make was displayed (MES condition); and 2) no advice was displayed (NOM condition). Behavioral results showed a significant effect of expert advice. Specifically, probability weighting functions changed in the direction of the expert's advice. This was paralleled by neural activation patterns. Brain activations showing significant correlations with valuation (parametric modulation by value of lottery/sure win) were obtained in the absence of the expert's advice (NOM) in intraparietal sulcus, posterior cingulate cortex, cuneus, precuneus, inferior frontal gyrus and middle temporal gyrus. Notably, no significant correlations with value were obtained in the presence of advice (MES). These findings were corroborated by region of interest analyses. Neural equivalents of probability weighting functions showed significant flattening in the MES compared to the NOM condition in regions associated with probability weighting, including anterior cingulate cortex, dorsolateral PFC, thalamus, medial occipital gyrus and anterior insula. Finally, during the MES condition, significant activations in temporoparietal junction and medial PFC were obtained. CONCLUSIONS/SIGNIFICANCE: These results support the hypothesis that one effect of expert advice is to "offload" the calculation of value of decision options from the individual's brain. PMID- 19308262 TI - Prevalence of obesity in school-going children of Karachi. AB - BACKGROUND: Obesity is an emerging problem in Pakistan. The authors sought to determine prevalence of obesity and malnutrition in school-going children, from grades 6(th) to 8(th) of different schools of Karachi and assess associations that affect the weight of the children. METHODOLOGY/PRINCIPAL FINDINGS: A cross sectional study design with children studying in grades 6(th) to 8(th) grade, in different schools of Karachi. We visited 10 schools of which 4 consented; two subsidized government schools and two private schools. A questionnaire was developed in consultation with a qualified nutritionist. Height and weight were measured on calibrated scales. A modified BMI criterion for Asian populations was used. Data was collected from 284 students. Of our sample, 52% were found to be underweight whereas 34% of all the children were normal. Of the population, 6% was obese and 8% overweight. Of all obese children, 70% belonged to the higher socio-economic status (SES) group, while of the underweight children, 63.3% were in the lower SES. Amongst obese children in our study, 65% ate meat every day, compared to 33% of normal kids. CONCLUSION: Obesity and undernutrition co-exist in Pakistani school-children. Our study shows that socio-economic factors are important since obesity and overweight increase with SES. Higher SES groups should be targeted for overweight while underweight is a problem of lower SES. Meat intake and lack of physical activity are some of the other factors that have been highlighted in our study. PMID- 19308263 TI - C5a enhances dysregulated inflammatory and angiogenic responses to malaria in vitro: potential implications for placental malaria. AB - BACKGROUND: Placental malaria (PM) is a leading cause of maternal and infant mortality. Although the accumulation of parasitized erythrocytes (PEs) and monocytes within the placenta is thought to contribute to the pathophysiology of PM, the molecular mechanisms underlying PM remain unclear. Based on the hypothesis that excessive complement activation may contribute to PM, in particular generation of the potent inflammatory peptide C5a, we investigated the role of C5a in the pathogenesis of PM in vitro and in vivo. METHODOLOGY AND PRINCIPAL FINDINGS: Using primary human monocytes, the interaction between C5a and malaria in vitro was assessed. CSA- and CD36-binding PEs induced activation of C5 in the presence of human serum. Plasmodium falciparum GPI (pfGPI) enhanced C5a receptor expression (CD88) on monocytes, and the co-incubation of monocytes with C5a and pfGPI resulted in the synergistic induction of cytokines (IL-6, TNF, IL-1beta, and IL-10), chemokines (IL-8, MCP-1, MIP1alpha, MIP1beta) and the anti angiogenic factor sFlt-1 in a time and dose-dependent manner. This dysregulated response was abrogated by C5a receptor blockade. To assess the potential role of C5a in PM, C5a plasma levels were measured in malaria-exposed primigravid women in western Kenya. Compared to pregnant women without malaria, C5a levels were significantly elevated in women with PM. CONCLUSIONS AND SIGNIFICANCE: These results suggest that C5a may contribute to the pathogenesis of PM by inducing dysregulated inflammatory and angiogenic responses that impair placental function. PMID- 19308264 TI - MicroRNA-199b-5p impairs cancer stem cells through negative regulation of HES1 in medulloblastoma. AB - BACKGROUND: Through negative regulation of gene expression, microRNAs (miRNAs) can function in cancers as oncosuppressors, and they can show altered expression in various tumor types. Here we have investigated medulloblastoma tumors (MBs), which arise from an early impairment of developmental processes in the cerebellum, where Notch signaling is involved in many cell-fate-determining stages. MBs occur bimodally, with the peak incidence seen between 3-4 years and 8 9 years of age, although it can also occur in adults. Notch regulates a subset of the MB cells that have stem-cell-like properties and can promote tumor growth. On the basis of this evidence, we hypothesized that miRNAs targeting the Notch pathway can regulated these phenomena, and can be used in anti-cancer therapies. METHODOLOGY/PRINCIPAL FINDINGS: In a screening of MB cell lines, the miRNA miR 199b-5p was seen to be a regulator of the Notch pathway through its targeting of the transcription factor HES1. Down-regulation of HES1 expression by miR-199b-5p negatively regulates the proliferation rate and anchorage-independent growth of MB cells. MiR-199b-5p over-expression blocks expression of several cancer stem cell genes, impairs the engrafting potential of MB cells in the cerebellum of athymic/nude mice, and of particular interest, decreases the MB stem-cell-like (CD133+) subpopulation of cells. In our analysis of 61 patients with MB, the expression of miR-199b-5p in the non-metastatic cases was significantly higher than in the metastatic cases (P = 0.001). Correlation with survival for these patients with high levels of miR-199b expression showed a positive trend to better overall survival than for the low-expressing patients. These data showing the down-regulation of miR-199b-5p in metastatic MBs suggest a potential silencing mechanism through epigenetic or genetic alterations. Upon induction of de-methylation using 5-aza-deoxycytidine, lower miR-199b-5p expression was seen in a panel of MB cell lines, supported an epigenetic mechanism of regulation. Furthermore, two cell lines (Med8a and UW228) showed significant up-regulation of miR-199b-5p upon treatment. Infection with MB cells in an induced xenograft model in the mouse cerebellum and the use of an adenovirus carrying miR-199b-5p indicate a clinical benefit through this negative influence of miR-199b-5p on tumor growth and on the subset of MB stem-cell-like cells, providing further proof of concept. CONCLUSIONS/SIGNIFICANCE: Despite advances in our understanding of the pathogenesis of MB, one-third of these patients remain incurable and current treatments can significantly damage long-term survivors. Here we show that miR-199b-5p expression correlates with metastasis spread, identifying a new molecular marker for a poor-risk class in patients with MB. We further show that in a xenograft model, MB tumor burden can be reduced, indicating the use of miR199b-5p as an adjuvant therapy after surgery, in combination with radiation and chemotherapy, for the improvement of anti-cancer MB therapies and patient quality of life. To date, this is the first report that expression of a miRNA can deplete the tumor stem cells, indicating an interesting therapeutic approach for the targeting of these cells in brain tumors. PMID- 19308265 TI - Quinolone-Based Third-Line Therapy for Helicobacter pylori Eradication. AB - Currently, a standard third-line therapy for Helicobacter pylori (H. pylori) eradication remains to be established. Quinolones show good oral absorption, no major side effects, and marked activity against H. pylori. Several authors have studied quinolone-based third-line therapy and reported encouraging results, with the reported H. pylori cure rates ranging from 60% to 84%. Resistance to quinolones is easily acquired, and the resistance rate is relatively high in countries with a high consumption rate of these drugs. We recently reported a significant difference in the eradication rate obtained between patients infected with gatifloxacin-susceptible and gatifloxacin-resistant H. pylori, suggesting that the selection of quinolones for third-line therapy should be based on the results of drug susceptibility testing. As other alternatives of third-line rescue therapies, rifabutin-based triple therapy, high-dose proton pump inhibitor/amoxicillin therapy and furazolidone-based therapy have been suggested. PMID- 19308266 TI - Menthol Enhances an Antiproliferative Activity of 1alpha,25-Dihydroxyvitamin D(3) in LNCaP Cells. AB - 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], the most active form of vitamin D(3), and its analogues have therapeutic benefits for prostate cancer treatment. However, the development of hypercalcemia is an obstacle to clinical applications of 1alpha,25(OH)(2)D(3) for cancer therapy. In this study, we provide evidence that menthol, a key component of peppermint oil, increases an anti-proliferation activity of 1alpha,25(OH)(2)D(3) in LNCaP prostate cancer cells. We found that menthol per se does not exhibit antiproliferative activity, but it is able to enhance 1alpha,25(OH)(2)D(3)-mediated growth inhibition in LNCaP cells. Fluorometric assays using Fura-2 showed that 1alpha,25(OH)(2)D(3) does not induce acute Ca(2+) response, whereas menthol evokes an increase in [Ca(2+)](i), which suggests that cross-talks of menthol-induced Ca(2+) signaling with 1alpha,25(OH)(2)D(3)-mediated growth inhibition pathways. In addition, Western blot analysis revealed that 1alpha,25(OH)(2)D(3) and menthol cooperatively modulate the expression of bcl-2 and p21 which provides the insight into the molecular mechanisms underlying the enhanced 1alpha,25(OH)(2)D(3) mediated growth inhibition by menthol. Thus, our findings suggest that menthol may be a useful natural compound to enhance therapeutic effects of 1alpha,25(OH)(2)D(3). PMID- 19308267 TI - Intestinal inflammation in rats induces metallothionein in colonic submucosa. AB - The aim of the current study was to determine if induction of metallothionein (MT) via acute or chronic dietary zinc supplementation attenuates intestinal inflammation, and to investigate the relationship with site-specific intestinal MT determined by immunolocalization. Growing rats were assigned to zinc-deficient (ZD), acute zinc-treated (ZT), pair-fed, control or chronic Zn-supplemented (ZS) groups. Half the rats in each dietary group received 5% dextran sulphate sodium (DSS) in their drinking water for 4 days. DSS treatment produced acute intestinal inflammation in the colon only, however, dietary zinc deficiency, acute zinc treatment or chronic zinc supplementation did not alter the severity of ulceration. Serum zinc concentrations were attenuated in the DSS-challenged ZT and ZS groups suggesting that zinc was being utilized in some capacity in response to inflammation. DSS-challenge induced MT immunostaining in the colonic submucosa, however, MT was not associated with histological improvements in the present study. The site-specific MT induction in colonic submucosa during intestinal inflammation requires further clarification as a component of the host defense. PMID- 19308268 TI - Beneficial Effects of Cocoa in Perivascular Mato Cells of Cerebral Arterioles in SHR-SP (Izm) Rats. AB - As previously reported, the cerebral arterioles are surrounded by unique perivascular Mato cells. They contain many inclusion bodies rich in hydrolytic enzymes, and have strong uptake capacity. They are thus considered scavenger cells of vascular and neural tissues in steady-state. In this study, employing hypertensive SHR-SP (Izm) rats, the viability of Mato cells was investigated. In hypertensive rats, the capacity for uptake of horse radish peroxidase (HRP) and the activity of acid phosphatase (ACPase) of Mato cells were markedly reduced, and on electron-microscopic examination Mato cells were found to include heterogeneous contents and appeared electron-dense and degenerated. Vascular cells exhibited some signs of pathology. However, in hypertensive rats fed chow containing 0.25% cocoa, the uptake capacity and ACPase activity of Mato cells for HRP were enhanced, and on electron-microscopic examination Mato cells appeared healthy, with mitochondria with nearly normal profiles. Signs of pathology in vascular cells were also decreased. Superoxides may impair Mato cells and vascular cells. PMID- 19308269 TI - Dose-dependent effects of barley cooked with white rice on postprandial glucose and desacyl ghrelin levels. AB - White rice is an indispensable staple food in Japan, although it is a high glycemic index food. The objective of this study was to estimate how barley cooked with white rice might affect postprandial glucose, insulin and desacyl ghrelin concentrations as well as fullness. The study was conducted in randomized crossover design with nine healthy subjects. Blood glucose, insulin, free fatty acid and desacyl ghrelin concentrations and subjective levels of fullness and hunger were measured for 240 min after intake of glucose, white rice, 30% rolled barley (30BAR), 50% rolled barley (50BAR) and 100% rolled barley (100BAR) containing 75 g of available carbohydrate. Postprandial glucose and insulin levels were suppressed by intake of 30BAR, 50BAR and 100BAR comparing with those of white rice. Area under the curves of plasma glucose and insulin concentrations was reduced by barley intake in a dose-dependent manner. Although plasma desacyl ghrelin levels decreased postprandially, the degree of reduction was suppressed by barley intake in a dose-dependent manner. Postprandial desacyl ghrelin levels can be a sensitive biomarker of carbohydrate metabolism. The combination of white rice with barley plays a beneficial role in preventing and treating type 2 diabetes, obesity and other metabolic diseases. PMID- 19308270 TI - Effect of Obstructive Jaundice and Nitric Oxide on the Profiles of Intestinal Bacterial Flora in Wild and iNOS Mice. AB - We previously reported that the plasma level of endotoxin and colonic expression of IgA in the mouse increased with obstructive jaundice induced by bile duct ligation (BDL). To elucidate the mechanism of the BDL-induced increase, we analyzed the effect of BDL on intestinal flora in wild type and inducible nitric oxide synthase (iNOS)-deficient mice (iNOS(-/-)) using the terminal restriction fragment length polymorphism analysis (T-RFLP) and 16S rDNA clone libraries. The amounts of bacterial DNA detected in fecal samples from both animal groups pretreated with antibiotics were extremely low as compared with untreated groups. We found that the profiles of enteric bacteria changed markedly after BDL. The bacterial composition is significantly different between not only wild type and iNOS(-/-) mice but also those before and after BDL, respectively. Among enteric bacteria examined, Lactobacillus murinus was found to increase markedly after BDL in rectum of both animal groups. However, Escherichia coli markedly increased after BDL in the iNOS(-/-) mice. These findings suggest that profiles of enteric flora change markedly in animals during obstructive jaundice by some mechanism that is affected by bile constituents and iNOS-derived NO. PMID- 19308271 TI - Effects of Short-Term Dietary Change from High-Carbohydrate Diet to High-Fat Diet on Storage, Utilization, and Fatty Acid Composition of Rat Muscle Triglyceride during Swimming Exercise. AB - The purpose was to examine the effects of a 3-day dietary change from a high carbohydrate (C) to high-fat (F) diet on muscle triglyceride (MTG) storage and utilization during the swimming exercise in rats. Rats were meal-fed on either the F diet or the C diet for 11 days. For an additional 3 days, half of the rats in each group were fed the same diets and the other rats were switched to counterpart diets. On the final day, half of the rats in each group were killed before the exercise and the others were killed after the exercise. Serum concentrations of glucose and free fatty acids (FFA) were higher in the post exercise groups than in the pre-exercise groups. The tissue glycogen contents were lower in the post-exercise groups. However, the MTG contents and fatty acid (FA) compositions were not influenced by the exercise and dietary change. The F diet increased the FFA concentration and slightly increased the MTG content. Moreover, the dietary FA composition influenced the FA composition of the MTG. These results suggest that the exercise did not affect the contents and FA composition of MTG, but that the F diet had an effect on the MTG contents and FA composition. PMID- 19308272 TI - Effect of Nitric Oxide on the Oxygen Metabolism and Growth of E. faecalis. AB - Gastro-intestinal mucosal cells have a potent mechanism to eliminate a variety of pathogens using enzymes that generate reactive oxygen species and/or nitric oxide (NO). However, a large number of bacteria survive in the intestine of human subjects. Enterococcus faecalis (E. faecalis) is a Gram-positive bacterium that survives not only in the intestinal lumen but also within macrophages generating NO. It has been reported that E. faecalis generated the superoxide radical (O(2) (-)). To elucidate the role of O(2) (-) and NO in the mechanism for the pathogen surviving in the intestine and macrophages, we studied the role and metabolism of O(2) (-) and NO in and around E. faecalis. Kinetic analysis revealed that E. faecalis generated 0.5 micromol O(2) (-)/min/10(8) cells in a glucose-dependent manner as determined using the cytochrome c reduction method. The presence of NOC12, an NO donor, strongly inhibited the growth of E. faecalis without affecting in the oxygen consumption. However, the growth rate of NOC12-pretreated E. faecalis in NO-free medium was similar to that of untreated cells. Western blotting analysis revealed that the NOC12-treated E. faecalis revealed a large amount of nitrotyrosine-posititive proteins; the amounts of the modified proteins were higher in cytosol than in membranes. These observations suggested that O(2) (-) generated by E. faecalis reacted with NO to form peroxinitrite (ONOO(-)) that preferentially nitrated tyrosyl residues in cytosolic proteins, thereby reversibly inhibited cellular growth. Since E. faecalis survives even within macrophages expressing NO synthase, similar metabolism of O(2) (-) and NO may occur in and around phagocytized macrophages. PMID- 19308273 TI - Oxidative stress biomarkers and lifestyles in Japanese healthy people. AB - The urinary concentrations of 8-isoprostane and 8-hydroxy-2'-deoxyguanosine (8 OHdG), which are biomarkers of oxidative stress, were measured in 677 Japanese people without any diseases, and their correlations with lifestyle facotrs, lifestyle-related blood biochemical parameters, and dietary intake of antioxidative vitamins were investigated. The mean urinary concentration of 8 isoprostane and 8-OHdG was 0.58 ng/mg creatinine and 8.43 ng/mg creatinine, respectively. Mean urinary 8-isoprostane was significantly different in terms of age, gender, smoking and alcohol consumption but not different in terms of body mass index (BMI) and exercise. By multiple regression analysis, urinary 8 isoprostane was significantly influenced by smoking and age. On the other hand, mean urinary 8-OHdG showed differences only by age group. Multiple regression analysis revealed that urinary 8-OHdG was significantly influenced by age, smoking, body weight, levels of high-sensitivity C-reactive protein (Hs-CRP) and low density lipoprotein-cholesterol in females, although it was significantly influenced by body weight in males. The present study shows that urinary 8 isoprostane is associated with lipid peroxidation related-lifestyles such as smoking, and urinary 8-OHdG is associated with arteriosclerosis related-factors such as Hs-CRP. Our findings suggest that 8-isoprostane and 8-OHdG appear to be prospective biomarkers for early prediction of lifestyle related-disease risk at the population level. PMID- 19308274 TI - The Expression of Murine Double Minute 2 (MDM2) on Helicobacter pylori-Infected Intestinal Metaplasia and Gastric Cancer. AB - The overexpression of murine double minute 2 (MDM2) is found in several human tumors, and increased expression of MDM2 inactivates the apoptotic and cell cycle arrest function of p53. Interleukin-16 (IL-16) is a pleiotrophic cytokine and the properties of IL-16 suggest that it involve in the pathophysiological process of chronic inflammatory diseases. In this study, we investigated the expression of MDM2 in intestinal metaplasia and gastric cancer as well as the effect of H. pylori infection and IL-16 on epithelial cell proliferation and MDM2 expression in gastric cells in vitro. The expression of MDM2 on gastric biopsies was studied immunohistochemistry. AGS cells were incubated with a combination of IL-16 and Helicobacter pylori (H. pylori). Gastric epithelial cell proliferation was studied by BrdU uptake and the expressions of MDM2 were studied by ELISA. There was no significant difference on the expression of MDM2 between with and without H. pylori infected chronic gastritis. In H. pylori infected gastric mucosa; the MDM2 expression was higher on intestinal metaplasia and gastric cancer than chronic gastritis. IL-16 administration was increased MDM2 expression and cell proliferation on AGS cells, which was decreased by H. pylori infection. In conclusion, the expression of MDM2 in long-term H. pylori infected gastric mucosa may indicate a risk for carcinogenesis. IL-16 secretion in H. pylori infected mucosa is one of the factors for gastric cancer. The expression of MDM2 on mucosa can be a mediator for gastric cancer. PMID- 19308275 TI - Ablation techniques for mahaim fiber tachycardia. AB - Mahaim fiber exhibits atrio-ventricular node like properties and generally is localized at the lateral aspect of the tricuspid annulus. Of the varying methods for localization, ablation at the site of Mahaim potential is the most accepted and successful method. Radiofrequency ablation of Mahaim fiber has high success rates. PMID- 19308276 TI - Morphology of current of injury does not predict long term active fixation ICD lead performance. AB - BACKGROUND: Currents of injury (COI) have been associated with improved lead performance during perioperative measurements in pacemaker and ICD implants. Their relevance on long term lead stability remains unclear. METHODS: Unipolar signals were recorded immediately after active fixation ICD lead positioning, blinded to the implanting surgeon. Signals were assigned to prespecified COI types by two independent investigators. Sensing, pacing as well as changes requiring surgical intervention were prospectively investigated for 3 months. RESULTS: 105 consecutive ICD lead implants were studied. All could be assigned to a particular COI with 48 type 1, 43 type 2 and 14 type 3 signals. Pacing impedance at implant was 703.8+/-151.6 Ohm with a significant COI independent drop within the first week. Sensing was 10.6mV+/- 3.7mV and pacing threshold at implant was 0.8+/-0.3mV at 0.5ms at implant. There was no significant difference between COI groups at implant and during a 3 months follow up regarding sensing, pacing nor surgical revisions. CONCLUSIONS: Three distinct patterns of unipolar endocardial potentials were observed in active fixation ICD lead implant, but COI morphology did not predict lead performance after 3 months. PMID- 19308277 TI - Reliability of a novel intracardiac electrogram method for AV And VV delay optimization and comparability to echocardiography procedure for determining optimal conduction delays in CRT patients. AB - BACKGROUND: Echocardiography is widely used to optimize CRT programming. A novel intracardiac electrogram method (IEGM) was recently developed as an automated programmer-based method, designed to calculate optimal atrioventricular (AV) and interventricular (VV) delays and provide optimized delay values as an alternative to standard echocardiographic assessment. OBJECTIVE: This study was aimed at determining the reliability of this new method. Furthermore the comparability of IEGM to existing echocardiographic parameters for determining optimal conduction delays was verified. METHODS: Eleven patients (age 62.9+/- 8.7; 81% male; 73% ischemic), previously implanted with a cardiac resynchronisation therapy defibrillator (CRT-D) underwent both echocardiographic and IEGM-based delay optimization. RESULTS: Applying the IEGM method, concordance of three consecutively performed measurements was found in 3 (27%) patients for AV delay and in 5 (45%) patients for VV delay. Intra-individual variation between three measurements as assessed by the IEGM technique was up to 20 ms (AV: n=6; VV: n=4). E-wave, diastolic filling time and septal-to-lateral wall motion delay emerged as significantly different between the echo and IEGM optimization techniques (p < 0.05). The final AV delay setting was significantly different between both methods (echo: 126.4 +/- 29.4 ms, IEGM: 183.6 +/- 16.3 ms; p < 0.001; correlation: R = 0.573, p = 0.066). VV delay showed significant differences for optimized delays (echo: 46.4 +/- 23.8 ms, IEGM: 10.9 +/- 7.0 ms; p <0.01; correlation: R = -0.278, p = 0.407). CONCLUSION: The automated programmer-based IEGM-based method provides a simple and safe method to perform CRT optimization. However, the reliability of this method appears to be limited. Thus, it remains difficult for the examiner to determine the optimal hemodynamic settings. Additionally, as there was no correlation between the optimal AV- and VV-delays calculated by the IEGM method and the echo optimization, the use of the IEGM method and the comparability to the echo has not been definitely clarified. PMID- 19308278 TI - Suggestions to evaluate whether T-wave alternans is T-wave amplitude dependent. PMID- 19308279 TI - Letter by Bohora regarding article, "Brugada-type electrocardiographic pattern induced by electrocution". PMID- 19308281 TI - Unusual site of permanent pacing: a case report. AB - Subclavian route is a standard way of performing a permanent pacemaker. However in cases with superior vena caval or bilateral subclavian occlusion and bilateral infection of pacemaker site, alternative site is warranted. Epicardial route needs general anesthesia and has its own problems. Iliofemoral route has been used previously but has more lead related problems and patient discomfort. Here we are reporting a case in which transiliac pacing was done due to both right and left pacemaker site active infection and to avoid the chance of lead dislodgement, an alpha loop was made in the right atrium. PMID- 19308282 TI - Looks like VT But Isn't--successful ablation of a left free wall accessory pathway with Mahaim-like properties. AB - It was long believed that Mahaim pathways represented nodo-fascicular or nodo ventricular connections. However, this misconception was challenged when patients underwent surgical or catheter ablation of the AV node but remained pre-excited. Electrophysiology (EP) studies showed these pathways to be right sided decrementally conducting atrio-fascicular accessory pathways with the atrium forming a part of the antidromic tachycardia circuit. Mahaim pathways are usually reported to occur on the right side. We report a patient who presented with a broad complex tachycardia thought to be ventricular tachycardia; however during EP study this was shown to be an antidromic atrioventricular tachycardia utilising a left free wall pathway that demonstrated 'Mahaim-like' properties and was successfully ablated. The pathway was shown to have long conduction times with no retrograde conduction, had an effective refractory period longer than the AV node and its conduction was only evident during antidromic AVRT. It also had a decremental antegrade property and was responsive to intravenous adenosine. These 'Mahaim-like' features are very unusual and rarely reported on the left side. PMID- 19308283 TI - Ablating the ventricular insertion of atrio-fascicular mahaim fiber: could be performed safely? AB - We report a patient who underwent radiofrequency ablation of the distal insertion of an atrio-fascicular accessory pathway with decremental properties because of inability to map a suitable potential alongside the tricuspid annulus. Small, discrete potentials resembling those of Purkinje fiber were found at right ventricular apex. All these potentials showed early activation during tachycardia preceding the QRS onset of various degree. Pace mapping helped to localize the presumed main distal insertion of the atrio-fascicular accessory pathway in a region where damage of the His-purkinje system may ensue. This case report describes catheter ablation of an atriofascicular accessory pathway by targeting its distal (ventricular) insertion site. PMID- 19308284 TI - A narrow QRS complex Tachycardia with an apparently concentric retrograde atrial activation sequence. AB - The retrograde atrial activation sequence constitutes an initial important clue to elucidate the tachycardia mechanism during diagnostic electrophysiological testing in patients with supraventricular tachycardia. However, in some cases its correct analysis is challenging. PMID- 19308285 TI - Wolff-Parkinson-White syndrome: where is the pathway? AB - A 31-year old male presented with atrial fibrillation and ventricular preexcitation that was positive in leads V1-V4, negative in lead II, and positive in lead AVR. The patient was cardioverted and invasive electrophysiologic study was performed. Based on the ECG findings, the coronary sinus and its branches were interrogated during orthodromic atrioventricular reentrant tachycardia. The earliest local activation was seen in the true coronary sinus lumen at the bifurcation of the posterolateral branch. Radiofrequency energy application at this area led to loss of preexcitation. When localizing left septal and posterior accessory pathways, ventricular preexcitation that is both negative in II and positive in AVR has been shown in previous studies to be highly sensitive and specific for a subepicardial location. Therefore, investigation of the coronary sinus and its branches may allow for effective ablation without the need for left ventricular access. PMID- 19308286 TI - Acetylation of FoxO1 activates Bim expression to induce apoptosis in response to histone deacetylase inhibitor depsipeptide treatment. AB - Histone deacetylase (HDAC) inhibitors have been shown to induce cell cycle arrest and apoptosis in cancer cells. However, the mechanisms of HDAC inhibitor induced apoptosis are incompletely understood. In this study, depsipeptide, a novel HDAC inhibitor, was shown to be able to induce significant apoptotic cell death in human lung cancer cells. Further study showed that Bim, a BH3-only proapoptotic protein, was significantly upregulated by depsipeptide in cancer cells, and Bim's function in depsipeptide-induced apoptosis was confirmed by knockdown of Bim with RNAi. In addition, we found that depsipeptide-induced expression of Bim was directly dependent on acetylation of forkhead box class O1 (FoxO1) that is catalyzed by cyclic adenosine monophosphate-responsive element-binding protein binding protein, and indirectly induced by a decreased four-and-a-half LIM-domain protein 2. Moreover, our results demonstrated that FoxO1 acetylation is required for the depsipeptide-induced activation of Bim and apoptosis, using transfection with a plasmid containing FoxO1 mutated at lysine sites and a luciferase reporter assay. These data show for the first time that an HDAC inhibitor induces apoptosis through the FoxO1 acetylation-Bim pathway. PMID- 19308287 TI - Quantitative physiology of the precancerous cervix in vivo through optical spectroscopy. AB - Cervical cancer is the second most common female cancer worldwide. The ability to quantify physiological and morphological changes in the cervix is not only useful in the diagnosis of cervical precancers but also important in aiding the design of cost-effective detection systems for use in developing countries that lack well-established screening and diagnostic programs. We assessed the capability of a diffuse reflectance spectroscopy technique to identify contrasts in optical biomarkers that vary with different grades of cervical intraepithelial neoplasia (CIN) from normal cervical tissues. The technology consists of an optical probe and an instrument (with broadband light source, dispersive element, and detector), and a Monte Carlo algorithm to extract optical biomarker contributions including total hemoglobin (Hb) concentration, Hb saturation, and reduced scattering coefficient from the measured spectra. Among 38 patients and 89 sites examined, 46 squamous normal sites, 18 CIN 1, and 15 CIN 2(+) sites were included in the analysis. Total Hb was statistically higher in CIN 2(+) (18.3 +/- 3.6 microM, mean +/- SE) compared with normal (9.58 +/- 1.91 microM) and CIN 1 (12.8 +/- 2.6 microM), whereas scattering was significantly reduced in CIN 1 (8.3 +/- 0.8 cm(-1)) and CIN 2(+) (8.6 +/- 1.0 cm(-1)) compared with normal (10.2 +/- 1.1 cm(-1)). Hemoglobin saturation was not significantly altered in CIN 2(+) compared with normal and CIN 1. The difference in total Hb is likely because of stromal angiogenesis, whereas decreased scattering can be attributed to breakdown of collagen network in the cervical stroma. PMID- 19308288 TI - Staphylococcal superantigen-like 10 inhibits CXCL12-induced human tumor cell migration. AB - PURPOSE: Tumor cell migration and metastasis share many similarities with leukocyte trafficking, which is critically regulated by chemokines and their receptors. CXCR4 is the most widely expressed chemokine receptor in many different types of cancer and has been linked to tumor dissemination and poor prognosis. Several CXCR4 antagonists have been synthesized. A totally novel approach to discover chemokine receptor antagonists is the use of bacteria. Bacteria produce chemokine receptor inhibitors to prevent neutrophil extravasation and migration toward the infection site to escape clearance by innate immune cells. The aim of the current study was to find and identify the mechanism of a bacterial protein that specifically targets CXCR4, a chemokine receptor shared by neutrophils and cancer cells. EXPERIMENTAL DESIGN: Several staphylococcal proteins were screened for their capacity to prevent binding of a function-blocking antibody against CXCR4. RESULTS: Staphylococcal superantigen like 10 was found to bind CXCR4 expressed on human T acute lymphoblastic leukemia, lymphoma, and cervical carcinoma cell lines. It potently inhibited CXCL12-induced calcium mobilization and cell migration. CONCLUSIONS: Staphylococcal superantigen-like 10 is a potential lead in the development of new anticancer compounds preventing metastasis by targeting CXCR4. PMID- 19308289 TI - PDCD5 interacts with Tip60 and functions as a cooperator in acetyltransferase activity and DNA damage-induced apoptosis. AB - Tip60 is a histone acetyltransferase (HAT) involved in the acetyltransferase activity and the cellular response to DNA damage. Here, we show that programmed cell death 5 (PDCD5), a human apoptosis-related protein, binds to Tip60 and enhances the stability of Tip60 protein in unstressed conditions. The binding amount of PDCD5 and Tip60 is significantly increased after UV irradiation. Further, PDCD5 enhances HAT activity of Tip60 and Tip60-dependent histone acetylation in both basal and UV-induced levels. We also find that PDCD5 increases Tip60-dependent K120 acetylation of p53 and participates in the p53 dependent expression of apoptosis-related genes, such as Bax. Moreover, we demonstrate the biological significance of the PDCD5-Tip60 interaction; that is, they function in cooperation to accelerate DNA damage-induced apoptosis and knockdown of PDCD5 or Tip60 impairs their apoptosis-accelerating activity, mutually. Consistent with this, PDCD5 levels increase significantly on DNA damage in U2OS cells, as does Tip60. Together, our findings indicate that PDCD5 may play a dual role in the Tip60 pathway. Specifically, under normal growth conditions, PDCD5 contributes to maintaining a basal pool of Tip60 and its HAT activity. After DNA damage, PDCD5 functions as a Tip60 coactivator to promote apoptosis. PMID- 19308290 TI - Discovery of fully human anti-MET monoclonal antibodies with antitumor activity against colon cancer tumor models in vivo. AB - The receptor tyrosine kinase MET is a major component controlling the invasive growth program in embryonic development and in invasive malignancies. The discovery of therapeutic antibodies against MET has been difficult, and antibodies that compete with hepatocyte growth factor (HGF) act as agonists. By applying phage technology and cell-based panning strategies, we discovered two fully human antibodies against MET (R13 and R28), which synergistically inhibit HGF binding to MET and elicit antibody-dependent cellular cytotoxicity. Cell based phosphorylation assays demonstrate that R13 and R28 abrogate HGF-induced activation of MET, AKT1, ERK1/2, and HGF-induced migration and proliferation. FACS experiments suggest that the inhibitory effect is mediated by "locking" MET receptor in a state with R13, which then increases avidity of R28 for the extracellular domain of MET, thus blocking HGF binding without activating the receptor. In vivo studies demonstrate that the combination of R13/28 significantly inhibited tumor growth in various colon tumor xenograft models. Inhibition of tumor growth was associated with induction of hypoxia. Global gene expression analysis shows that inhibition of HGF/MET pathway significantly upregulated the tumor suppressors KLF6, CEACAM1, and BMP2, the negative regulator of phosphatidylinositol-3-OH-kinase PIK3IP1, and significantly suppressed SCF and SERPINE2, both enhancers of proliferation and invasiveness. Moreover, in an experimental metastasis model, R13/28 increased survival by preventing the recurrence of otherwise lethal lung metastases. Taken together, these results underscore the utility of a dual-antibody approach for targeting MET and possibly other receptor tyrosine kinases. Our approach could be expanded to drug discovery efforts against other cell surface proteins. PMID- 19308291 TI - Activin receptor signaling regulates prostatic epithelial cell adhesion and viability. AB - Mutational changes coupled with endocrine, paracrine, and/or autocrine signals regulate cell division during carcinogenesis. The hormone signals remain undefined, although the absolute requirement in vitro for fetal serum indicates the necessity for a fetal serum factor(s) in cell proliferation. Using prostatic cancer cell (PCC) lines as a model of cancer cell proliferation, we have identified the fetal serum component activin A and its signaling through the activin receptor type II (ActRII), as necessary, although not sufficient, for PCC proliferation. Activin A induced Smad2 phosphorylation and PCC proliferation, but only in the presence of fetal bovine serum (FBS). Conversely, activin A antibodies and inhibin A suppressed FBS-induced PCC proliferation confirming activin A as one of multiple serum components required for PCC proliferation. Basic fibroblast growth factor was subsequently shown to synergize activin A induced PCC proliferation. Inhibition of ActRII signaling using a blocking antibody or antisense-P decreased mature ActRII expression, Smad2 phosphorylation, and the apparent viability of PCCs and neuroblastoma cells grown in FBS. Suppression of ActRII signaling in PCC and neuroblastoma cells did not induce apoptosis as indicated by the ratio of active/inactive caspase 3 but did correlate with increased cell detachment and ADAM-15 expression, a disintegrin whose expression is strongly correlated with prostatic metastasis. These findings indicate that ActRII signaling is required for PCC and neuroblastoma cell viability, with ActRII mediating cell fate via the regulation of cell adhesion. That ActRII signaling governs both cell viability and cell adhesion has important implications for developing therapeutic strategies to regulate cancer growth and metastasis. PMID- 19308292 TI - NHERF-1: modulator of glioblastoma cell migration and invasion. AB - The invasive nature of malignant gliomas is a clinical problem rendering tumors incurable by conventional treatment modalities such as surgery, ionizing radiation, and temozolomide. Na(+)/H(+) exchanger regulatory factor 1 (NHERF-1) is a multifunctional adaptor protein, recruiting cytoplasmic signaling proteins and membrane receptors/transporters into functional complexes. This study revealed that NHERF-1 expression is increased in highly invasive cells that reside in the rim of glioblastoma multiforme (GBM) tumors and that NHERF-1 sustains glioma migration and invasion. Gene expression profiles were evaluated from laser capture-microdissected human GBM cells isolated from patient tumor cores and corresponding invaded white matter regions. The role of NHERF-1 in the migration and dispersion of GBM cell lines was examined by reducing its expression with small-interfering RNA followed by radial migration, three dimensional collagen dispersion, immunofluorescence, and survival assays. The in situ expression of NHERF-1 protein was restricted to glioma cells and the vascular endothelium, with minimal to no detection in adjacent normal brain tissue. Depletion of NHERF-1 arrested migration and dispersion of glioma cell lines and caused an increase in cell-cell cohesiveness. Glioblastoma multiforme cells with depleted NHERF-1 evidenced a marked decrease in stress fibers, a larger cell size, and a more rounded shape with fewer cellular processes. When NHERF-1 expression was reduced, glioma cells became sensitized to temozolomide treatment resulting in increased apoptosis. Taken together, these results provide the first evidence for NHERF-1 as a participant in the highly invasive phenotype of malignant gliomas and implicate NHERF-1 as a possible therapeutic target for treatment of GBM. PMID- 19308293 TI - Upregulated INHBA expression may promote cell proliferation and is associated with poor survival in lung adenocarcinoma. AB - INTRODUCTION: The expression, mechanisms of regulation, and functional impact of INHBA (activin A) in lung adenocarcinoma (AD) have not been fully elucidated. METHODS: INHBA expression was examined in 96 lung samples (86 ADs, 10 normal lung) using oligonucleotide microarrays and 187 lung samples (164 ADs, 6 bronchioalveolar carcinomas, and 17 normal lung) using immunohistochemistry. The proliferation of AD cell lines H460 and SKLU1 was examined with WST-1 assays after treatment with recombinant activin A, follistatin, and INHBA-targeting small-interfering RNA. Cells were also treated with 5-aza-2' deoxycytidine and trichostatin A to investigate the role of epigenetic regulation in INHBA expression. RESULTS: Primary ADs expressed 3.1 times more INHBA mRNA than normal lung. In stage I AD patients, high levels of primary tumor INHBA transcripts were associated with worse prognosis. Immunohistochemistry confirmed higher inhibin betaA protein expression in ADs (78.7%) and bronchioalveolar carcinomas (66.7%) compared with normal lung (11.8%). H460 and SKLU1 demonstrated increased proliferation when treated with exogenous activin A and reduced proliferation when treated with follistatin or INHBA-targeting small-interfering RNA. INHBA mRNA expression in H460 cells was upregulated after treatment with trichostatin A and 5-aza-2' deoxycytidine. CONCLUSIONS: INHBA is overexpressed in AD relative to controls. Inhibin betaA may promote cell proliferation, and its overexpression is associated with worse survival in stage I AD patients. In addition, overexpression of INHBA may be affected by promoter methylation and histone acetylation in a subset of lung ADs. PMID- 19308294 TI - Loss of p53 induces tumorigenesis in p21-deficient mesenchymal stem cells. AB - There is growing evidence about the role of mesenchymal stem cells (MSCs) as cancer stem cells in many sarcomas. Nevertheless, little is still known about the cellular and molecular mechanisms underlying MSCs transformation. We aimed at investigating the role of p53 and p21, two important regulators of the cell cycle progression and apoptosis normally involved in protection against tumorigenesis. Mesenchymal stem cells from wild-type, p21(-/-)p53(+/+), and p21(-/-)p53(+/-) mice were cultured in vitro and analyzed for the appearance of tumoral transformation properties after low, medium, and high number of passages both in vitro and in vivo. Wild-type or p21(-/-)p53(+/+) MSCs did not show any sign of tumoral transformation. Indeed, after short-term in vitro culture, wild-type MSCs became senescent, and p21(-/-)p53(+/+) MSCs showed an elevated spontaneous apoptosis rate. Conversely, MSCs carrying a mutation in one allele of the p53 gene (p21(-/-)p53(+/-) MSCs) completely lost p53 expression after in vitro long term culture. Loss of p53 was accompanied by a significant increase in the growth rate, gain of karyotypic instability, loss of p16 expression, and lack of senescence response. Finally, these cells were able to form fibrosarcomas partially differentiated into different mesenchymal lineages when injected in immunodeficient mice both after subcutaneous and intrafemoral injection. These findings show that MSCs are very sensitive to mutations in genes involved in cell cycle control and that these deficiencies can be at the origin of some mesodermic tumors. PMID- 19308295 TI - Depletion of serotonin and selective inhibition of 2B receptor suppressed tumor angiogenesis by inhibiting endothelial nitric oxide synthase and extracellular signal-regulated kinase 1/2 phosphorylation. AB - The effects of serotonin (5-HT) on tumor growth are inconsistent. We investigated whether a decreased level of 5-HT affected tumor growth using 5-HT transporter knockout (5-HTT(-/-)) mice, which showed 5-HT depletion. When cancer cells were injected subcutaneously into both 5-HTT(-/-) and 5-HTT(+/+) mice, the tumor growth was markedly attenuated in 5-HTT(-/-) mice. Serotonin levels in the blood, forebrain, and tumors of 5-HTT(-/-) mice bearing tumors were significantly smaller than those of their 5-HTT(+/+) littermates. However, 5-HT did not increase cancer cells' proliferation in vitro. When we applied 5-HTT inhibitors to the wild mice bearing tumors, they did not inhibit tumor growth. The endothelial nitric oxide synthase (eNOS) expressions in tumors were reduced in 5 HTT(-/-) mice compared with 5-HTT(+/+) mice. Stimulations with 5-HT (1-50 microM) induced eNOS expressions in human umbilical vein endothelial cell (HUVEC) in a concentration-dependent manner. When we measured activations of multiple signaling pathways by using a high-throughput phosphospecific antibodies platform, 5-HT stimulated the extracellular signal-regulated kinase 1/2 (ERK1/2) in HUVEC. Moreover, we found that the physiological level of 5-HT induced phosphorylation of both ERK1/2 and eNOS in HUVEC. Human umbilical vein endothelial cell expressed both 5-HT(2B) and 5-HT(2C) receptors. SB204741, a specific 5-HT(2B) receptor inhibitor, blocked 5-HT-induced ERK1/2 and eNOS phosphorylations, whereas RS102221, a specific 5-HT(2C) receptor inhibitor, did not in HUVEC. SB204741 reduced microvessel density in tumors and inhibited the proliferation of HUVEC in vitro. These results suggest that regulation of 5-HT and 5-HT receptors, especially the 5-HT(2B) receptor, may serve as a therapeutic strategy in cancer therapy. PMID- 19308296 TI - Peak pressure and tidal volume are affected by how the neonatal self-inflating bag is handled. AB - OBJECTIVE: To evaluate how different ways of handling the neonatal self-inflating bag influences peak pressure and tidal volume. METHODS: This is an experimental study involving 141 different professionals (physicians, resident physicians, physiotherapists, nurses and nursing technicians), who ventilated an artificial lung, adjusted to simulate the lung of a term neonate, using a self-inflating bag. Each professional handled the ventilator in five different ways: a) using both hands (10 fingers); and, with only one hand, b) five fingers, c) four fingers, d) three fingers and e) two fingers. Peak pressure and tidal volume data were recorded by the artificial lung equipment. RESULTS: Both variables showed high variability, from 2.5 to 106.3 cmH2O (mean = 39.73 cmH2O; 95%CI 37.32-42.13) for peak pressure, and from to 4 to 88 mL (mean = 39.56 mL; 95%CI 36.86-42.25) for tidal volume. There was no significant influence of the profession on any of the variables (p > 0.05). However, bag handling significantly influenced both peak pressure and tidal volume (p < 0.0001), which were higher when the operator used both hands. CONCLUSION: The results indicate that most professionals delivered excessively high peak pressures and tidal volumes, which could increase the risk of barotrauma and volutrauma, especially when both hands were used to ventilate. On the other hand, a small number of professionals delivered insufficient pressure and volume for adequate lung expansion and ventilation. The delivery of inadequate ventilation was not dependent on profession. PMID- 19308297 TI - Interaction between postural risk factors and job strain on self-reported musculoskeletal symptoms among users of video display units: a three-year prospective study. AB - OBJECTIVE: This study investigated a possible interaction between postural risk factors and job strain on the incidence proportion of self-reported musculoskeletal symptoms in the shoulder-neck, lower back and upper limbs regions. METHODS: A cohort of white-collar workers (N=2431) was assessed with a self-administered questionnaire regarding postural risk factors and job strain at work. After a three-year follow-up, the six-month incidence proportion of musculoskeletal symptoms in the three body regions was measured with a modified version of the Nordic questionnaire. The analyses were stratified for gender. Interaction was defined as a departure from the addition of effects of individual risk factors, and its importance was estimated from the attributable proportion due to interaction and its 95% confidence interval (95% CI). RESULTS: A significant attributable proportion of 0.80 (95% CI 0.23-1.37) due to interaction between postural risk factors and job strain was observed for men in the lower back region. An indication of interaction was found for women with attributable proportions due to interaction of 0.44 (95% CI -0.06-0.94), 0.27 (95% CI -0.34 0.88) and 0.36 (95% CI -0.33-1.05) for the shoulder-neck, lower back, and upper limbs regions, respectively. CONCLUSIONS: The simultaneous presence of postural risk factors and job strain seems to increase the pathogenic effect of each exposure on the incidence proportion of musculoskeletal symptoms. This interaction effect is important for work intervention practices as success in decreasing any of these two risk factors could have the additional benefit of reducing up to 80% of new cases of musculoskeletal symptoms among participants exposed to both risk factors. PMID- 19308298 TI - Changes in stress and coping from a randomized controlled trial of a three-month stress management intervention. AB - OBJECTIVES: To investigate whether a group-based stress management intervention, based on principles from cognitive behaviour therapy, can reduce stress and alter coping strategies in an occupationally diverse population with extensive symptoms of work-related stress. METHODS: Using a randomized wait list control design, 102 participants were divided into two groups: intervention and wait list control (WLC). The intervention was a three-month group-based stress management program. Outcomes measures were the Perceived Stress Scale (PSS-10, range 0-40 points) and five dimensions from the Brief COPE questionnaire (range 2-8 points) at baseline and three-, six- and nine-months follow-up. Data were analyzed with a univariate analysis of variance. RESULTS: On the PSS-10 from baseline to three months, the intervention group changed -6.45 (95% CI -8.25--4.64) points, compared to -1.12 (95% CI -2.94-0.70) points in the WLC group. The between-groups difference was 5.32 (95% CI -7.89--2.76) points, equalling a standardized mean difference of 0.84 (95% CI -1.27--0.42) favouring the intervention. One coping dimension, positive reframing, differed between the two groups. Here the intervention group changed -0.86 (95% CI -1.25--0.48) points from baseline to three months, compared to -0.18 (-0.58-0.22) points in the WLC-group. We found a between-groups difference of -0.67 (95% CI -1.24--0.11) points, equalling a standardized mean difference of -0.48 (95% CI -0.89--0.07) favouring the intervention. The gains achieved during treatment were maintained when followed up three months later. CONCLUSIONS: Treatment is superior to the control condition in positively affecting perceived stress and positive reframing. When followed up, the gains achieved are maintained. PMID- 19308299 TI - Joint toxicity evaluation and QSAR modeling of aromatic amines and phenols to bacteria. AB - Acute toxicity of aromatic amines and phenols and their mixtures to bacteria in natural waters was determined and the median inhibition concentration IC(50) values for single compounds and IC(50mix) values for binary and multiple mixtures were obtained. Based on the quantitative structure-activity relationship model for single chemical toxicity, a two-descriptor model accounting for n octanol/water partition coefficient (log P(mix)) and the energy of the lowest unoccupied molecular orbital (E(LUMOmix)) was developed for the toxicity of a mixture: log(1/IC(50mix) = 0.326 log P(mix) - 0.660E(LUMOmix) + 3.323 (n = 32, R(2) = 0.834). This model can be used successfully to predict the toxicity of a mixture, whether binary mixtures or multiple mixtures of three or four chemicals are used as predictors. PMID- 19308300 TI - Peripheral arterial disease and osteoporosis in older adults: the Rancho Bernardo Study. AB - SUMMARY: We examined the association between peripheral arterial disease (PAD) and bone health in 1,332 adults. We found a weak association between PAD and osteoporosis and bone loss only in women, but the association was not independent of age. PAD was not associated with fractures in this community-based population. INTRODUCTION: Increased rates of osteoporosis have been reported in patients with cardiovascular disease, suggesting a link between osteoporosis and atherosclerosis. METHODS: We examined the association between PAD and bone health in 1,332 adults who attended a research visit in 1992-1996, when the ankle brachial index (ABI), bone mineral density (BMD), and spine X-rays were obtained. A total of 837 participants attended a follow-up visit in 1997-2000. RESULTS: PAD defined by an ABI < or = 0.90 was present in 15.4% of the women and 13.3% of the men. Prevalence of osteoporosis was significantly higher in women with PAD compared to women without PAD (p < 0.05). During an average 4-year follow-up, women with PAD had a significantly higher rate of bone loss than women without PAD (p = 0.05). The associations were no longer significant after age adjustment. In men, PAD was not associated with osteoporosis, but men with PAD had lower BMD at the femoral neck than men without PAD (p = 0.03). PAD was not associated with osteoporotic fractures in either sex. CONCLUSION: We found a weak and age dependent association between PAD and osteoporosis in women but not men. PAD was not associated with fractures in this community-based population. PMID- 19308301 TI - Vitamin D status and falls, frailty, and fractures among postmenopausal Japanese women living in Hawaii. AB - Vitamin D status and its relationship to physical performance, falls, and fractures in 495 postmenopausal women of Japanese ancestry in Hawaii were investigated. The mean 25-hydroxyvitamin D (25-OHD) was 31.94 ng/mL. No significant association of 25-OHD was demonstrated with most outcomes, possibly due to higher 25-OHD levels in this population. INTRODUCTION: In this study, we investigated vitamin D status and its relationship to physical performance, muscle strength, falls, and fractures in postmenopausal Japanese females living in Hawaii. METHODS: Of 510 community-dwelling women who participated in the eighth examination of the Hawaii Osteoporosis Study, 495 were included in these analyses. Multivariate regression models were used to evaluate the relationship of 25-OHD (D(3) and total) to eight performance-based measurements, 12 activities of daily living (ADLs), and muscle strength (grip, triceps, and quadriceps). Logistic regression analyses were performed to evaluate the relationship of 25 OHD to falls, vertebral fractures, and non-vertebral fractures. RESULTS: The mean total 25-OHD was 31.94 +/- 9.46 ng/mL; 44% of subjects had values <30 ng/mL, while none had values <10-12 ng/mL. There was little evidence of seasonal variation. Among performance-based measures, ADLs, and strength tests, only quadriceps strength was significantly associated with total 25-OHD (p = 0.0063) and 25-OHD(3) (p = 0.0001). No significant association of 25-OHD was found with vertebral or non-vertebral fractures, or incidence of one or more falls. CONCLUSIONS: Lack of serum 25-OHD relationship with falls and fractures or most physical performance measures in this study may be related to the low prevalence of very low 25-OHD levels in this population. PMID- 19308302 TI - Determinants of bone mass and size in 7-year-old former term, late-preterm, and preterm boys. AB - When compared, full-term prepubertal boys had greater regional bone size, higher total body (TB) bone mineral content (BMC), and regional bone density than preterm boys but higher TB bone content area and regional BMC than late-preterm boys. Implications include follow-up bone assessment and preterm formula feeding in late-preterm boys. INTRODUCTION: This study was conducted to determine whether there are differences in bone mass and size among prepubertal boys born preterm (PT; < or =34 weeks gestation), late-preterm (LP; >34 and < or =37 weeks gestation), and at term (>37 weeks gestation) and to identify factors that are associated with bone mass and size in these children. METHODS: Total body (TB), spine and hip dual energy X-ray absorptiometry and tibia peripheral quantitative computed tomography measures were obtained on 24 boys aged 5.7 to 8.3 years. RESULTS: In multiple regression analysis adjusting for current weight, height, age, and jump power, term boys had greater cortical thickness (p = 0.03) and area (p = 0.01), higher trabecular volumetric bone mineral density (p = 0.05), TB bone mineral content (BMC; p = 0.007), and hip areal bone mineral density (aBMD; p = 0.01) than PT boys, and higher TB BMC (p = 0.01), TB bone area (p = 0.03), hip BMC (p = 0.02) and aBMD (p = 0.01), and femoral neck BMC (p = 0.05) and aBMD (p = 0.02) than LP boys. There were no differences in activity measures among gestation groups and no group-by-activity interactions. CONCLUSION: Term boys have greater bone size and mass than PT boys and higher bone mass than LP boys at several bone sites. Activity measures did not differ among gestation groups and did not explain bone differences. PMID- 19308303 TI - Reliability of Tuffier's line evaluated on cadaver specimens. AB - The aim was to evaluate the reliability of Tuffier's line usually used as sole method to identify lumbar spinous process for a correct needle placement. Fifty eight cadaver specimens were placed in a lateral position and a flexion in the lumbar spine performed to achieve a neutralization of the lumbar lordosis. The iliac crests were palpated and the lumbar spinous process marked on the intercristal line with a pin; all specimens were dissected and the marked spinous process documented. The center of the L4 spinous process was hit in 24 male (41.38%) and 10 female (17.24%) specimens. In only two female specimens (3.45%), the upper vertebra was reached; a pin placed in L3 was not found in male cadavers. The inferior edge of the L4 spinous process was hit in male 4 times (6.90%) and in female cadavers 12 times (20.69%). In the fifth lumbar spinous process, pins were placed five times in female cadavers (8.62%) and in only one male cadaveric specimen (3.72%). In conclusion, the accuracy of the focused lumbar spinous process depends on the right bedding and the orientation of the given landmarks, so Tuffier's line stays the most important tool for anesthetists if palpation is performed very precisely. PMID- 19308304 TI - Anatomical insight into the normal and abnormal branching pattern of the popliteal artery. PMID- 19308305 TI - The efficacy of natalizumab in patients with relapsing multiple sclerosis: subgroup analyses of AFFIRM and SENTINEL. AB - The AFFIRM and SENTINEL studies showed that natalizumab was effective both as monotherapy and in combination with interferon beta (IFNbeta)-1a in patients with relapsing multiple sclerosis (MS). Further analyses of AFFIRM and SENTINEL data were conducted to determine the efficacy of natalizumab in prespecified patient subgroups according to baseline characteristics: relapse history 1 year before randomization (1, 2, > or = 3), Expanded Disability Status Scale score (< or = 3.5, > 3.5), number of T2 lesions (< 9, > or = 9), presence of gadolinium enhancing (Gd+) lesions (0, > or = 1), age (< 40, > or = 40) and gender (male, female). A post hoc analysis was conducted to determine the efficacy of natalizumab in patients with highly active disease (i. e., > or = 2 relapses in the year before study entry and > or = 1 Gd+ lesion at study entry). In both AFFIRM and SENTINEL studies natalizumab reduced the annualized relapse rates across all subgroups (except the small subgroups with < 9 baseline T2 lesions) over 2 years. In AFFIRM, natalizumab significantly reduced the risk of sustained disability progression in most subgroups. In SENTINEL, natalizumab significantly reduced the risk of sustained disability progression in the following subgroups: > or = 9 T2 lesions at baseline, > or = 1 Gd+ lesions at baseline, female patients and patients < 40 years of age. Natalizumab reduced the risk of disability progression by 64 % and relapse rate by 81 % in treatment- naive patients with highly active disease and by 58 % and 76 %, respectively, in patients with highly active disease despite IFNbeta-1a treatment. These results indicate that natalizumab is effective in reducing disability progression and relapses in patients with relapsing MS, particularly in patients with highly active disease. PMID- 19308306 TI - Natural history comparisons of primary and secondary progressive multiple sclerosis reveals differences and similarities. AB - BACKGROUND: Similarities in the onset age of progression in secondary-progressive (SP) and primary-progressive multiple sclerosis (PPMS) have been previously reported. However, with longer follow-up, more relapsing-remitting (RRMS) patients reach SPMS, such that the baseline characteristics, including age at progression may shift. We aimed to examine how this phenomenon impacts on demographic and clinical comparisons made between PP and SPMS. METHODS AND RESULTS: Patients with definite MS, onset by July 1988 and > or = 1 Expanded Disability Status Scale (EDSS) score were selected from the British Columbia-wide MS database (n = 2837). Of these, 353 (12.4 %) had PPMS and 1445/2484 (58.2 %) of the RRMS population reached SPMS at study close (July 2003). Females predominated in the SPMS population regardless of follow-up time (p < or = 0.032). From Kaplan Meier analysis (all RR, SP and PP patients considered), the estimated median onset age of progression was greater in SPMS (49.0 years; 95 % CI: 48.3-49.7) than PPMS (41.0 years; 95 % CI: 39.7-42.4), p < 0.0005. If the RR patients (who had not developed SPMS) were excluded, median age of onset of SPMS was still greater (43.1 years (95 % CI: 42.3-43.9, p < 0.0005). CONCLUSIONS: Although there were some similarities between SPMS and PPMS, the former had a later onset age in our British Columbian MS cohort. PMID- 19308307 TI - Apolipoprotein E genotype as a risk factor for susceptibility to and dementia in Parkinson's disease. AB - Further to the well-established association between apolipoprotein E (APOE) and Alzheimer's disease, this gene has also been implicated in both susceptibility to, and dementia in, Parkinson's disease (PD). However studies to date have produced contradictory findings. We conducted a case-control study in a population of 528 PD patients and 512 healthy controls and found no significant difference in allele or genotype distribution of APOE between the two groups. An updated meta-analysis showed a modest increase of APOE-epsilon2 carriers amongst PD patients compared to controls [P = 0.017, OR = 1.16 (95 % CI 1.03-1.31)]. 107 of our patients were incident cases participating in a population-based epidemiological study. Longitudinal follow-up of this cohort over a mean of 5.0 +/- 0.7 years from diagnosis revealed no significant impact of APOE-epsilon4 carrier status on risk of dementia or rate of cognitive decline. An updated meta analysis indicated an over-representation of APOE-epsilon4 carriers amongst PD dementia compared to non dementia cases [OR 1.74 (1.36-2.23), P = 1 x 10(-4)], although small sample sizes, heterogeneity of odds ratios and publication bias may have confounded this finding. In conclusion, our study does not support previously reported associations between APOE genotype and susceptibility to, or cognitive decline in, PD. An updated meta-analysis indicates any association with PD susceptibility is at most modest, an observation with important implications for further study of this issue. Large scale longitudinal studies would be best placed to further evaluate any impact of APOE genotype on cognitive decline in PD. PMID- 19308308 TI - Predicting recovery after intracerebral hemorrhage--an external validation in patients from controlled clinical trials. AB - BACKGROUND: An early and reliable prognostic indication in stroke patients is potentially useful for initiation of individual treatment and for informing patients and relatives. We recently developed a regression model as well as a simple 11-point predictive score (Essen ICH score) for functional recovery within three months after acute intracerebral hemorrhage (ICH) based on age and the National Institutes of Health Stroke Scale (NIH-SS). Here, we demonstrate the applicability of our models in an independent sample of ICH patients from controlled clinical trials. METHODS: The prognostic models were used to predict functional recovery in 564 patients from the Virtual International Stroke Trials Archive (VISTA). Furthermore, we tried to improve the accuracy by re-calibration and estimating new model parameters. FINDINGS: The logistic regression model and the Essen ICH score were able to correctly classify 77.5 % and 76.4 % of patients, respectively. Re-calibration and novel estimation of parameters yielded only a slight improvement of overall predictive accuracy. INTERPRETATION: For acute ICH patients included in controlled trials, our predictive models based on age and the NIH-SS correctly predict functional recovery after three months and could be useful for future trial design. PMID- 19308309 TI - Utility of the new Japanese severity score and indications for special therapies in acute pancreatitis. AB - BACKGROUND: The Japanese severity score (JSS) for acute pancreatitis was revised in 2008. As special therapies for severe acute pancreatitis (SAP), continuous regional arterial infusion of protease inhibitor and antibiotics (CRAI) and enteral nutrition (EN) are now utilized in Japan. We investigated the usefulness of the new JSS and the indications for CRAI and EN based on the new JSS. METHODS: We assessed the new JSS in 138 patients with SAP according to the previous Japanese criteria. Usefulness of the new JSS for the prediction of mortality rates was compared with conventional scoring systems by receiver-operator characteristic curve analysis. We analyzed the relationship between the new JSS and prognosis in patients with and without CRAI and EN, respectively. RESULTS: Forty-five patients (33%) were assessed as having mild acute pancreatitis, and 93 patients (67%) were assessed as having SAP. Their mortality rates were 7 and 40%, respectively. The area under the curve for the prediction of mortality rates with the new JSS was 0.822 and was the highest among conventional scoring systems. In patients with new JSS >or= 6, the mortality rate was lower in patients with CRAI than in patients without CRAI (P = 0.129). In patients with new JSS >or= 4, the mortality rate was lower in patients with EN than in patients without EN (P = 0.016). CONCLUSIONS: The new JSS is useful and easier to use for the prediction of prognosis compared to the conventional scoring systems. EN was effective in reducing the mortality rate in patients with a new JSS >or= 4. PMID- 19308310 TI - Meta-analysis: the effect of interferon on development of hepatocellular carcinoma in patients with chronic hepatitis B virus infection. AB - PURPOSE: In patients chronically infected with hepatitis B virus, interferon has been used for the purpose of viral suppression by loss of hepatitis B e antigen (HBeAg) with or without seroconversion to antibody to HBeAg (anti-HBe). However, discussion about the effect of interferon on the development of hepatocellular carcinoma (HCC) has been controversial. METHODS: We conducted a meta-analysis of published studies. Eight studies were retrieved (1,303 patients), including two randomized controlled trials (RCTs) and six non-RCTs (553 patients received interferon treatment). RESULTS: The pooled estimate of the preventive effect of treatment was significantly in favor of interferon (risk difference -5.0%; 95% CI -9.4 to -0.5, P = 0.028). By subgroup analyses, the preventive effect of interferon treatment was shown in the Asian population (risk difference -8.5%; 95%CI -13.6 to -3.6, P = 0.0012), the population with the incidental rate of HCC >or=10% if untreated with interferon (risk difference -9.4%; 95%CI -14.2 to -4.6, P = 0.0001), and the population with the proportion of HBeAg-positive patients to the study population >or=70% (RD -6.0%; 95%CI -11.8 to -0.2, P = 0.043). However, the preventive effect of interferon was not shown in the European population, the population with a lower incidental rate of HCC if untreated with interferon, and the population with the lower proportion of HBeAg-positive patients to the study population. An evaluation using the Begg method indicated no evidence of publication bias. CONCLUSIONS: Interferon treatment is considered to restrain HCC development in patients with chronic hepatitis B virus infection, especially in HBeAg-positive Asians. PMID- 19308311 TI - Gastric emptying of liquid and solid meals at various temperatures: effect of meal temperature for gastric emptying. AB - BACKGROUND: Patients with functional dyspepsia frequently show delayed gastric emptying, and dietary advice is frequently given for its improvement. If meal temperature influences gastric emptying, advice regarding the meal temperature may become a possible component of dietary therapy. However, little information exists concerning the thermal effect of meals on gastric emptying. The aim of this study was to determine the thermal effect of liquid and solid meals on gastric emptying. METHODS: The gastric emptying of liquid and solid test meals was examined in healthy volunteers (liquid, n = 25, mean age = 35.7 +/- 9.6 years, male-to-female ratio = 22:3; solid, n = 25, mean age = 35.2 +/- 8.8 years, male-to-female ratio = 20:5). Gastric emptying after the ingestion of liquid or solid meals at three different temperatures (4, 37, and 60 degrees C) was investigated with the [(13)C]-labeled acetate breath test. The lag phase time (T (max-calc)) and the half-emptying time (T (1/2)) were calculated from the (13)CO(2) breath excretion curve as indices of gastric emptying. RESULTS: The values of T (max-calc) at 60 degrees C with both the liquid and solid meals were significantly smaller than those at 37 degrees C (P < 0.05). However, there was no difference in the T (1/2) values. In the analysis of the percent excretion of (13)CO(2) in 1 h (% dose/h) data with the liquid meal test in the earlier phase within 30 min, significantly larger values were found at 60 degrees C than at the other temperatures. These findings suggest that a hot meal significantly accelerates gastric emptying. CONCLUSIONS: Meal temperature may be considered as a component of dietary therapy for patients with functional dyspepsia. PMID- 19308312 TI - FibroScan and ultrasonography in the prediction of hepatic fibrosis in patients with chronic viral hepatitis. AB - BACKGROUND: The aim of this study was to assess the diagnostic performances of liver stiffness measurement (LSM), ultrasonography (US) and their combined use in predicting the extent of hepatic fibrosis. METHODS: Consecutive patients with chronic hepatitis B (HBV) or hepatitis C virus (HCV) infections, with indications for liver biopsy, were prospectively enrolled. LSM was performed on the same day as biopsy. US scores, including assessment of liver surface, liver parenchyma, intrahepatic vessels and spleen index, were used to assess the degree of hepatic fibrosis. The pathological findings were used as a reference standard and diagnostic accuracy was assessed and compared. RESULTS: Three-hundred and twenty patients, including 199 men and 121 women, with a mean age of 50.8 years, were analyzed. There were 214 (66.9%) HCV patients, 88 (27.5%) HBV patients and 18 (5.6%) patients with both HCV and HBV. LSM correlated significantly with the hepatic fibrosis (F) scores, necro-inflammatory activity and US scores in multivariate analysis. The diagnostic accuracy of LSM is significantly superior to US, and equal to combined LSM with US, in the prediction of all HCV-related fibrosis scores. The cut-off value of LSM is 6 kPa for diagnosing F > =1, with a positive predictive value of 91%. Also, the cut-off value is 12 kPa for the prediction of cirrhosis, with a negative predictive value of 94%. CONCLUSIONS: LSM is useful for predicting hepatic fibrosis and excluding cirrhosis. A combination of LSM and US does not improve the accuracy in assessing hepatic fibrosis. PMID- 19308313 TI - A simple shoot multiplication procedure using internode explants, and its application for particle bombardment and Agrobacterium-mediated transformation in watercress. AB - A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 muM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1-2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 muM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene. PMID- 19308314 TI - Full-length sequence analysis of a distinct isolate of Bidens mottle virus infecting sunflower in Taiwan. AB - The full-length genome of a potyvirus, previously known as sunflower chlorotic spot virus isolate SF-1 (SCSV-SF-1) which causes novel symptoms on sunflowers (Helianthus annuus), was sequenced and analyzed. The genome of SCSV-SF-1 is 9,741 nucleotides long, encoding a polyprotein of 3,071 amino acids containing the consensus motifs of potyviruses. Sequence comparison revealed that the 3' terminus of SCSV-SF-1 shared over 96% similarities with isolates of Bidens mottle virus (BiMoV). However, SCSV-SF-1 has a very narrow host range, excluding the diagnostic host species for BiMoV, Bidens pilosa and Zinnia elegans. Therefore, SCSV-SF-1 is a distinct isolate of BiMoV. This is the first report of the full length nucleotide sequence of BiMoV infecting sunflower in Taiwan. PMID- 19308315 TI - High dietary caffeine consumption is associated with a modest increase in headache prevalence: results from the Head-HUNT Study. AB - The aim of this study was to investigate the association between caffeine consumption and headache type and frequency in the general adult population. The results were based on cross-sectional data from 50,483 (55%) out of 92,566 invited inhabitants aged >or=20 years who participated in the Nord-Trondelag Health Survey. In the multivariate analyses, adjusting for age, gender, smoking, and level of education as confounding factors, a weak but significant association (OR = 1.16, 95% CI 1.09-1.23) was found between high caffeine consumption and prevalence of infrequent headache. In contrast, headache >14 days/month was less likely among individuals with high caffeine consumption compared to those with low caffeine consumption. The results may indicate that high caffeine consumption changes chronic headache into infrequent headache due to the analgesic properties of caffeine. Alternatively, chronic headache sufferers tend to avoid intake of caffeine to not aggravate their headaches, whereas individuals with infrequent headache are less aware that high caffeine use can be a cause. PMID- 19308316 TI - Lazy, dynamic or minimally recrudescent? On the elusive nature and location of the mycobacterium responsible for latent tuberculosis. AB - In the absence of symptoms characteristic of tuberculosis (TB), a condition termed clinical latency, diagnosis is currently impossible by detection of the microorganism itself and resorts to the demonstration of an immunological memory response to antigens of Mycobacterium tuberculosis (Mtb). Whether latency is synonymous to chronic persistent infection with viable Mtb in all instances has been difficult to establish. The physical and physiological state of Mtb during latency is much disputed: are organisms mostly dormant, in a nonreplicating state of persistence, and characterized by lipid inclusions and metabolic adaptation to hypoxia, or do they continue to replicate and sometimes even escape from the fringes of granulomatous lesions or alveolar epithelial cells into adjacent airways, thereby inducing recurring immune responses? The physical nature of Mtb during latency is important as it determines which antimicrobial agents may be used to kill it, which immunomodulating strategies (including post-exposure vaccines) may be appropriate to contain it, and which diagnostic measures may be most useful to discriminate latent from reactivating infection. Two major viewpoints exist: one argues that Mtb persists mostly in a lazy state within granulomatous lesions, but periodically recrudesces, and that there is considerable heterogeneity for different sites within the lesion and within the infected lung. Throughout latency, there is a dynamic immunological interplay between Mtb and the host, necessitating continuous recruitment of cells into the granuloma, and reactivation occurs when this dynamic cellular exchange becomes dysregulated. Another view holds that dormant Mtb reside within alveolar epithelial cells in the lung apices and in adipocytes, with reactivation being associated with the upregulation of resuscitation promoting factors within Mtb and the escape of newly dividing microorganisms into alveoli and bronchi in the form of lipid pneumonia. These views need not be mutually exclusive. However, if minimal intermittent recrudescence were to take place within the alveolar space, this would contradict the very definition of latency, which implies that no access of Mtb to the airways exists during latency. PMID- 19308317 TI - Tuberculosis: an old but still fascinating disease. PMID- 19308318 TI - A dynamic reinfection hypothesis of latent tuberculosis infection. AB - BACKGROUND: It has been traditionally postulated that individuals, once infected by Mycobacterium tuberculosis, will retain throughout their entire lifetime latent bacilli which will remain dormant in old lesions. This bacillus would then be the source of a later reactivation of active tuberculosis (TB), with the aid of resuscitation factors. Unfortunately, the presence of these bacilli can only be predicted by indirect immunological methods, such as the tuberculin skin test (TST) or T cell interferon-gamma release assays. Other evidence shows that a 9 month isoniazid treatment of TST+ individuals converting to TB reduces the incidence of TB by approximately 90%. QUESTIONS: Taking into account this widely accepted framework, I suggest that there are at least three relevant questions to answer: (1) How can dormant bacilli remain in the lungs for an entire lifetime, taking into account constant cellular turnover and the healing of damaged tissues? (2) What provides the resuscitation factor to dormant bacilli, assuming that these latent bacilli are indeed present inside old lesions? (3) Why can a 9 month treatment with isoniazid eliminate dormant bacilli? As isoniazid is active only against growing bacilli, and thus is only able to destroy them after reactivation of latent bacilli, this treatment should have to be provided for life if the traditionally accepted postulate is correct. HYPOTHESIS: For a better understanding of latent TB infection. I propose a hypothesis that describes a dynamic scenario of constant endogenous reinfection with M. tuberculosis which explains the efficacy of the current standard of treatment. If this hypothesis is true, new strategies for the management of TB may arise. PMID- 19308319 TI - Adherence to surgical site infection guidelines in Italian cardiac surgery units. AB - BACKGROUND: Data on the adherence to surgical site infection (SSI) prevention guidelines in Italian cardiac surgery units are lacking. METHODS: A multiple choice questionnaire, structured into eight sections following the Centers for Disease Control 1999 (CDC) guidelines, was prepared and sent to 24 surgical units participating in a national study group (GIS-InCard); this units perform over 20% of all cardiac surgical procedures in Italy. Answers were stratified based upon the evidence of the recommendations: grade IA (ten questions), grade IB (52 questions), grade II (11 questions), and no recommendation (seven questions). RESULTS: 17 of the 24 units (72%) returned the questionnaire. Adherence to grade IA recommendations was 69 +/- 34%, with five units (29%) showing a > or =80% adherence. Adherence to grade IB and II was 65 +/- 26% and 71 +/- 28%, respectively. Adherence did not vary significantly depending on the evidence of the recommendation, i.e., grade IA, IB or II (p = 0.72). Low adherence levels to grade I recommendations were observed on hair removal: (1) it was performed systematically in all male patients (0% adherence), (2) it was performed on the morning of the intervention in 29% of centers, and (3) the method of hair removal was adequate in 41% of cases. Despite 94% of units having written guidelines on antibiotic prophylaxis, only 65% administered antibiotic prophylaxis with the correct timing - i.e., on anesthesia induction. CONCLUSIONS: Adherence to CDC SSI guidelines in Italy is fair. The evidence of the recommendation does not influence adherence. Organizational improvements, especially those regarding hair removal and the timing of antibiotic prophylaxis, should be implemented in most hospitals. PMID- 19308321 TI - Evolutionary and functional epitopes of the Spatzle protein: new insights into activation of the Toll receptor. AB - Spatzle, a dimeric ligand, binds to the Drosophila Toll receptor and activates the signal pathway functioning in both embryonic patterning and innate immunity. Here, we used the evolutionary trace approach based on phylogenetic information to predict the evolutionary epitope of Spatzle and found that it mainly clusters in several adjacent loops of Spatzle far from the cystine-knot structural domain. We designed six mutants of Spatzle based on the evolutionary epitope and transfected them into a stable cell line expressing the luciferase reporter gene under the control of the drosomycin promoter. Luciferase assays showed that these mutants cannot significantly activate the drosomycin promoter, suggesting the involvement of these sites in binding of Spatzle to the Toll receptor. These data highlight the importance of the Trp-loop of the mushroom-shaped Spatzle dimer in Toll receptor activation and demonstrate that evolution-guided site-specific mutagenesis represents a useful and promising strategy for understanding the ligand-receptor interaction. PMID- 19308320 TI - Women facing HIV. Key question on women with HIV infection: Italian consensus workshop. AB - A panel of leading Italian specialists in infectious diseases, obstetrics and gynaecology met in a national consensus workshop on women facing HIV to review critical aspects and discuss recommendations for selected key questions on four issues: (1) women and highly active antiretroviral therapy (HAART): access to care and adherence to therapy, side effects and drug-drug interaction; (2) HIV infected pregnant women: prevention of mother to child transmission; (3) desire for children among women living with HIV: assisted reproduction; (4) sexually transmitted diseases and genital disturbances. The method of a nominal group meeting was used, and recommendations were graded for their strength and quality of evidence using a system based on the one adopted by the Infectious Diseases Society of America. Main conclusions are summarized and critically discussed, and some of the most recent data supporting recommendations are provided. PMID- 19308322 TI - Recent advances in understanding the structure and function of general transcription factor TFIID. AB - The general transcription factor TFIID is a macromolecular complex comprising the TATA-binding protein (TBP) and a set of 13-14 TBP associated factors (TAFs). This review discusses biochemical, genetic and electron microscopic data acquired over the past years that provide a model for the composition, organisation and assembly of TFIID. We also revisit ideas on how TFIID is recruited to the promoters of active and possibly repressed genes. Recent observations show that recognition of acetylated and methylated histone residues by structural domains in several TAFs plays an important role. Finally, we highlight several genetic studies suggesting that TFIID is required for initiation of transcription, but not for maintaining transcription once a promoter is in an active state. PMID- 19308323 TI - Roles of glial cells in synapse development. AB - Brain function relies on communication among neurons via highly specialized contacts, the synapses, and synaptic dysfunction lies at the heart of age-, disease-, and injury-induced defects of the nervous system. For these reasons, the formation-and repair-of synaptic connections is a major focus of neuroscience research. In this review, I summarize recent evidence that synapse development is not a cell-autonomous process and that its distinct phases depend on assistance from the so-called glial cells. The results supporting this view concern synapses in the central nervous system as well as neuromuscular junctions and originate from experimental models ranging from cell cultures to living flies, worms, and mice. Peeking at the future, I will highlight recent technical advances that are likely to revolutionize our views on synapse-glia interactions in the developing, adult and diseased brain. PMID- 19308324 TI - Functional aspects of protein flexibility. AB - Proteins are dynamic entities, and they possess an inherent flexibility that allows them to function through molecular interactions within the cell, among cells and even between organisms. Appreciation of the non-static nature of proteins is emerging, but to describe and incorporate this into an intuitive perception of protein function is challenging. Flexibility is of overwhelming importance for protein function, and the changes in protein structure during interactions with binding partners can be dramatic. The present review addresses protein flexibility, focusing on protein-ligand interactions. The thermodynamics involved are reviewed, and examples of structure-function studies involving experimentally determined flexibility descriptions are presented. While much remains to be understood about protein flexibility, it is clear that it is encoded within their amino acid sequence and should be viewed as an integral part of their structure. PMID- 19308326 TI - A completely in vitro system for obtaining scFv using mRNA display, PCR, direct sequencing, and wheat embryo cell-free translation. AB - Using mRNA display followed by in vitro sequencing and translation, a complete in vitro system for obtaining scFv has been developed. An mRNA display library for synthetic scFv was panned against human TNF receptor (TNFR). The nucleotide portion of the enriched molecules was subjected to limiting dilution, and PCR amplified. Three of the proteins encoded by the amplified fragments were synthesized in a wheat embryo (WE) cell-free system using a batch method. They were shown to bind TNFR by ELISA. One of their sequences was identified in vitro. The identified clone was further synthesized at approx. 0.5 mg/ml reaction mixture in a WE system with dialysis as a totally soluble protein. PMID- 19308325 TI - The multifaceted role of periostin in tumorigenesis. AB - Periostin, also called osteoblast-specific factor 2 (OSF-2), is a member of the fasciclin family and a disulfide-linked cell adhesion protein that has been shown to be expressed preferentially in the periosteum and periodontal ligaments, where it acts as a critical regulator of bone and tooth formation and maintenance. Furthermore, periostin plays an important role in cardiac development. Recent clinical evidence has also revealed that periostin is involved in the development of various tumors, such as breast, lung, colon, pancreatic, and ovarian cancers. Periostin interacts with multiple cell-surface receptors, most notably integrins, and signals mainly via the PI3-K/Akt and other pathways to promote cancer cell survival, epithelial-mesenchymal transition (EMT), invasion, and metastasis. In this review, aspects related to the function of periostin in tumorigenesis are summarized. PMID- 19308327 TI - DFT-based QSAR and QSPR models of several cis-platinum complexes: solvent effect. AB - Cytotoxic activities of cis-platinum complexes against parental and resistant ovarian cancer cell lines were investigated by quantitative structure-activity relationship (QSAR) analysis using density functional theory (DFT) based descriptors. The calculated parameters were found to increase the predictability of each QSAR model with incorporation of solvent effects indicating its importance in studying biological activity. Given the importance of logarithmic n octanol/water partition coefficient (log P(o/w)) in drug metabolism and cellular uptake, we modeled the log P(o/w) of 24 platinum complexes with different leaving and carrier ligands by the quantitative structure-property relationship (QSPR) analysis against five different concentrations of MeOH using DFT and molecular mechanics derived descriptors. The log P(o/w) values of an additional set of 20 platinum complexes were also modeled with the same descriptors. We investigated the predictability of the model by calculating log P(o/w) of four compounds in the test set and found their predicted values to be in good agreement with the experimental values. The QSPR analyses performed on 24 complexes, combining the training and test sets, also provided significant values for the statistical parameters. The solvent medium played an important role in QSPR analysis by increasing the internal predictive ability of the models. PMID- 19308328 TI - Pressure-dependent 13C chemical shifts in proteins: origins and applications. AB - Pressure-dependent (13)C chemical shifts have been measured for aliphatic carbons in barnase and Protein G. Up to 200 MPa (2 kbar), most shift changes are linear, demonstrating pressure-independent compressibilities. CH(3), CH(2) and CH carbon shifts change on average by +0.23, -0.09 and -0.18 ppm, respectively, due to a combination of bond shortening and changes in bond angles, the latter matching one explanation for the gamma-gauche effect. In addition, there is a residue specific component, arising from both local compression and conformational change. To assess the relative magnitudes of these effects, residue-specific shift changes for protein G were converted into structural restraints and used to calculate the change in structure with pressure, using a genetic algorithm to convert shift changes into dihedral angle restraints. The results demonstrate that residual (13)C alpha shifts are dominated by dihedral angle changes and can be used to calculate structural change, whereas (13)C beta shifts retain significant dependence on local compression, making them less useful as structural restraints. PMID- 19308329 TI - NMR investigations on residue level unfolding thermodynamics in DLC8 dimer by temperature dependent native state hydrogen exchange. AB - Understanding protein stability at residue level detail in the native state ensemble of a protein is crucial to understanding its biological function. At the same time, deriving thermodynamic parameters using conventional spectroscopic and calorimetric techniques remains a major challenge for some proteins due to protein aggregation and irreversibility of denaturation at higher temperature values. In this regard, we describe here the NMR investigations on the conformational stabilities and related thermodynamic parameters such as local unfolding enthalpies, heat capacities and transition midpoints in DLC8 dimer, by using temperature dependent native state hydrogen exchange; this protein aggregates at high (>65 degrees C) temperatures. The stability (free energy) of the native state was found to vary substantially with temperature at every residue. Significant differences were found in the thermodynamic parameters at individual residue sites indicating that the local environments in the protein structure would respond differently to external perturbations; this reflects on plasticity differences in different regions of the protein. Further, comparison of this data with similar data obtained from GdnHCl dependent native state hydrogen exchange indicated many similarities at residue level, suggesting that local unfolding transitions may be similar in both the cases. This has implications for the folding/unfolding mechanisms of the protein. PMID- 19308330 TI - Analytical solution to the coupled evolution of multidimensional NMR data. AB - A substantial time savings in the collection of multidimensional NMR data can be achieved by coupling the evolution of nuclei in the indirect dimensions. In order to save time, the sampling of the indirect dimensions is inherently incomplete. Therefore, many algorithms and samplings schemes have been developed aimed at separating the coevolved frequencies into analyzable data with limited artifacts. This paper extends the use of circulant matrices to describe coupled evolution with convolutions. By understanding the data in terms of convolutions, there is an exact solution to the inversion problem of extracting the orthogonal vectors from the coupled dimensions. Previously, this inversion problem has been solved using peak coordinates extracted from spectra. In contrast, the method described here uses spectra directly. This solution suggests a simple sampling scheme of collecting N orthogonal spectra, and N + 1 projections at specific projection angles, however, the theory developed can be extended generally to arbitrary projection angles. The circulant matrix methodology is demonstrated for simulated and real data. Further, an algorithm for separating overlapped signals in the detected dimension is presented. The algorithm involves the forward calculation of the coupled spectra from the orthogonal spectra, followed by back calculation of the orthogonal spectra from the coupled spectra, thus permitting rigorous cross-validation. This algorithm is shown to be robust in that erroneous solutions give rise to large artifacts. PMID- 19308331 TI - [The German Clinical Trials Register: reasons, general and technical aspects, international integration]. AB - In order to provide a central portal for information on clinical research in Germany and thus to facilitate the search of planned, ongoing and completed clinical trials, the German Clinical Trials Register (GermanCTR) was implemented in cooperation with the WHO's registries network. It is an open access online register of clinical trials conducted in Germany, which allows all users to search for, register and share information on clinical trials. The project is funded by the Federal Ministry of Education and Research and is implemented at the Institute for Medical Biometry and Medical Informatics of the University Medical Center Freiburg as a joint project of the Clinical Trials Center Freiburg and the German Cochrane Center. Since October 2008 the GermanCTR is an approved WHO Primary Registry and allows clinical trial registration in Germany according to the requirements of the International Committee of Medical Journal Editors (ICMJE). Reasons for a national trials register, general and technical aspects of implementing the GermanCTR as well as the national and international integration are described here. PMID- 19308332 TI - [UV Filters. State of the art]. AB - This article describes the basic principles of photoprotection with classic sunscreens containing chemical and/or physical UV filters and standard methods of sun protection factor (SPF) determination. In addition, the new governmental regulations and recommendations of the European Commission for photoprotection are presented and compared to regulations in other regions of the world. Finally, the efficacy of sunscreens in sunlight-induced non-sunburn effects, such as immune suppression and carcinogenesis are discussed. PMID- 19308333 TI - Early identification of co-occurring pain, depression and anxiety. AB - BACKGROUND: Depression and anxiety frequently co-occur with pain and may affect treatment outcomes. Early identification of these co-occurring psychiatric conditions during routine pain screening may be critical for optimal treatment. OBJECTIVE: To determine aspects of pain related to psychological distress, and, among distressed patients, to determine whether pain factors are related to provider identification of distress. DESIGN: Cross-sectional interview of primary care patients and their providers participating in a Veteran's Administration HELP-Vets study. SUBJECTS: A total of 528 predominately male Veterans MEASUREMENTS AND MAIN RESULTS: We measured self-reported pain, including a 0-10 numeric rating scale and interference items from the Brief Pain Inventory. To evaluate distress, brief indicators of depression, anxiety and PTSD were combined. A substantial number of patients had psychological distress (41%), which was even higher (62%) among patients with moderate-severe current pain. Only 29% of those with distress reported talking to their provider about emotional problems during their visit. In multivariate analyses, other pain factors related to distress included interference with enjoyment of life and relationships with others, pain in multiple locations and joint pains. Prior diagnoses of depression and anxiety were also related to current distress. Only prior diagnosis and patient reported headaches and sleep interference because of pain were related to provider identification of distress. CONCLUSIONS: VA patients with moderate-severe pain are at high risk for psychological distress, which often goes unrecognized. Providers need to be more vigilant to mental health problems in patients experiencing high pain levels. Targeted screening for co-occurring conditions is warranted. PMID- 19308334 TI - Not perfect, but better: primary care providers' experiences with electronic referrals in a safety net health system. AB - BACKGROUND: Electronic referrals can improve access to subspecialty care in safety net settings. In January 2007, San Francisco General Hospital (SFGH) launched an electronic referral portal that incorporated subspecialist triage, iterative communication with referring providers, and existing electronic health record data to improve access to subspecialty care. OBJECTIVE: We surveyed primary care providers (PCPs) to assess the impact of electronic referrals on workflow and clinical care. DESIGN: We administered an 18-item, web-based questionnaire to all 368 PCPs who had the option of referring to SFGH. MEASUREMENTS: We asked participants to rate time spent submitting a referral, guidance of workup, wait times, and change in overall clinical care compared to prior referral methods using 5-point Likert scales. We used multivariate logistic regression to identify variables associated with perceived improvement in overall clinical care. RESULTS: Two hundred ninety-eight PCPs (81.0%) from 24 clinics participated. Over half (55.4%) worked at hospital-based clinics, 27.9% at county funded community clinics, and 17.1% at non-county-funded community clinics. Most (71.9%) reported that electronic referrals had improved overall clinical care. Providers from non-county-funded clinics (AOR 0.40, 95% CI 0.14-0.79) and those who spent > or =6 min submitting an electronic referral (AOR 0.33, 95%CI 0.18 0.61) were significantly less likely than other participants to report that electronic referrals had improved clinical care. CONCLUSIONS: PCPs felt electronic referrals improved health-care access and quality; those who reported a negative impact on workflow were less likely to agree. While electronic referrals hold promise as a tool to improve clinical care, their impact on workflow should be considered. PMID- 19308335 TI - Do new drugs increase life expectancy? A critique of a Manhattan Institute paper. AB - A recent study published by the Manhattan Institute, "Why Has Longevity Increased More in Some States than in Others? The Role of Medical Innovation and Other Factors," purported to show that the more rapid adoption of new drugs has substantial benefits in the form of increased life expectancy, higher productivity and lower non-drug health care expenditures. This study has been cited as evidence supporting the more rapid acceptance of new drugs in Medicaid, Medicare, and other public programs and has helped to shape public debate on the value of new drugs. This analysis questions the key conclusions of the study. It points out that the key statistical regressions appear to be misspecified, since they show anomalies such as a negative correlation between income growth and life expectancy and find no relationship between education and productivity growth. Methodological flaws addressed include lack of adjustment for infant mortality rates; inadequate proxy measures of health status; lack of adjustment for ages of individuals and other sociodemographic factors; inherent problems with the definition of drug age, or 'vintage;' and the failure to consider reverse causation as an obvious explanation for several findings. The Manhattan Institute study does not provide reliable evidence for favoring adoption of newer drugs in either public or private health care programs. PMID- 19308336 TI - Who gets disease management? AB - BACKGROUND: Disease management (DM) has been promoted to improve health outcomes and lower costs for patients with chronic disease. Unfortunately, most of the studies that support claims of DM's success suffer from a number of biases, the most important of which is selection bias, or bias in the type of patients enrolling. OBJECTIVE: To quantify the differences between those who do and do not enroll in DM. DESIGN, SETTING, AND PARTICIPANTS: This was an observational study of the health care use, costs, and quality of care of 27,211 members of a large health insurer who were identified through claims as having asthma, diabetes, or congestive heart failure, were considered to be at high risk for incurring significant claims costs, and were eligible to join a disease management program involving health coaching. MEASUREMENTS: We used health coach call records to determine which patients participated in at least one coaching call and which refused to participate. We used claims data for the 12 months before the start of intervention to tabulate costs and utilization metrics. In addition, we calculated HEDIS quality scores for the year prior to the start of intervention. RESULTS: The patients who enrolled in the DM program differed significantly from those who did not on demographic, cost, utilization and quality parameters prior to enrollment. For example, compared to non-enrollees, diabetes enrollees had nine more prescriptions per year and higher HbA1c HEDIS scores (0.70 vs. 0.61, p < 0.001). CONCLUSIONS: These findings illuminate the serious problem of selection into DM programs and suggest that the effectiveness levels found in prior evaluations using methodologies that don't address this may be overstated. PMID- 19308338 TI - An in vitro study of electrically active hydroxyapatite-barium titanate ceramics using Saos-2 cells. AB - Electrically active ceramics are of interest as bone graft substitute materials. This study investigated the ferroelectric properties of hydroxyapatite-barium titanate (HABT) composites and the behaviour of osteoblast-like cells seeded on their surfaces. A piezoelectric coefficient (d(33)) of 57.8 pCN(-1) was observed in HABT discs prepared for cell culture. The attachment, proliferation, viability, morphology and metabolic activity of cells cultured on unpoled HABT were comparable to those observed on commercially available hydroxyapatite at all time points. No indication of the cytotoxicity of HABT was detected. At one day after seeding, cell attachment was modified on both the positive and negative surfaces of poled HABT. After longer incubations, all parameters observed were comparable on poled and unpoled ceramics. The results indicate that HABT ceramics are biocompatible in the short term in vitro and that further investigation of cell responses to these materials under mechanical load and at longer incubation times is warranted. PMID- 19308337 TI - Tea consumption and risk of type 2 diabetes: a meta-analysis of cohort studies. AB - BACKGROUND: Tea consumption has been extensively studied in relation to various diseases, several epidemiologic studies have been performed to investigate the association of tea consumption with type 2 diabetes; however, the results of these studies were not entirely consistent. OBJECTIVE: To conduct a meta-analysis of studies that assessed the association of tea consumption and the risk of type 2 diabetes. RESEARCH DESIGN AND METHODS: We performed a systematic literature search through November 2008 in PUBMED, MEDLINE, EMBASE, and Cochrane Database of Systematic Reviews. The search was limited to English-language studies. Studies were excluded if they were type 1 diabetes, animal studies. Nine cohort studies were identified by two authors, and summary relative risks (RRs) were calculated using a random-effects model. RESULTS: We identified nine cohort studies, including 324,141 participants and 11,400 incident cases of type 2 diabetes with follow-up ranging from 5 to 18 years. The summary adjusted RR did not show that tea consumption was associated with a reduced type 2 diabetes risk (RR, 0.96; 95% confidence interval (CI), 0.92-1.01). Evidence from the results of our stratified analyses revealed that tea consumption > or =4 cups per day (RR, 0.8; 95% CI, 0.7 0.93) might play a role in the prevention of type 2 diabetes. However, no statistically significant association was observed for sex and the follow-up durations stratified between tea consumption and type 2 diabetes. CONCLUSIONS: This meta-analysis indicates that tea consumption > or =4 cups per day may lower the risk of type 2 diabetes. PMID- 19308339 TI - On the robust parametric detection of EEG artifacts in polysomnographic recordings. AB - We present an open, parametric system for automatic detection of EEG artifacts in polysomnographic recordings. It relies on independent parameters reflecting the relative presence of each of the eight types of artifacts in a given epoch. An artifact is marked if any of these parameters exceeds a threshold. These thresholds, set for each parameter separately, can be adjusted via "learning by example" procedure (multidimensional minimization with computationally intensive cost function), which can be used to automatically tune the parameters to new types of datasets, environments or requirements. Performance of the system, evaluated on 103 overnight polysomnographic recordings, revealed concordance with decisions of human experts close to the inter-expert agreement. To make this statement well defined, we review the methodology of evaluation for this kind of detection systems. Complete source code is available from http://eeg.pl; a user friendly version with Java interface is available from http://signalml.org. PMID- 19308341 TI - [Stroke and hypertension]. AB - Arterial hypertension is the most important risk factor for stroke. Many interventional trials have unambiguously proven the benefit of antihypertensive therapy in both primary and secondary prevention for all age categories. No recommendation for any single antihypertensive substance for the primary prevention of stroke exists. Achieving the therapeutic goal (normotension) is the crucial factor. In most patients, multiple combinations of antihypertensive drugs are required to do this. For high-risk patients and in secondary prevention, substances inhibiting the renin-angiotensin-system, especially combined with calcium antagonists and indapamid, may be advantageous, while beta-blockers appear to be less well suited. In patients suffering from left-ventricular hypertrophy or atrial fibrillation, sartanes are the best-documented drug class. As TIA or stroke will often disturb the normal circadian rhythm of blood pressure and eliminate the usual night-time drop, monitoring of the therapeutic results must include ambulatory 24h measurements. The interrelation between vascular dementia and hypertension is by now also considered proven. An early start of antihypertensive treatment can prevent the development of dementia and impaired cognitive function. PMID- 19308345 TI - [Exercise and cellular adaptation of muscle]. AB - Resistance training and to a lesser extent endurance training are capable of enhancing protein synthesis in skeletal muscle via various signaling pathways. Additionally, the expression of muscle fiber types responds to different regimes of training stimuli and immobilization as characterized by changes in myosin heavy chain isoforms (I<-->IIA<-->IIX). Eccentric resistance training has been shown to be highly efficient in inducing sarcomeric protein assembly in the longitudinal orientation of muscle cells. However, concentric contractions lead to a hypertrophic response (increased fiber diameter) in muscle which can still be activated in old age. The central signaling pathway to mediate the elevation of protein synthesis in response to training is the mTOR pathway, which is also stimulated by free amino acids. Moreover, adaptation to endurance training is mediated by the calcium-calcineurin-NFATc1 pathway which is strongly activated by the calcium transients involved in the muscle contraction process. High contraction frequency and long duration of training sessions are essential for activation and maintenance of fiber type I expression as well as for induction of transformation of type II into type I fibers. Endurance training sessions should therefore be longer than 30 min and dominated by periods of high frequency contractions. A further factor in the muscular response to training includes the recruitment and integration of satellite cells into muscle fibers. Satellite cells can respond to muscular stretch, activity and injury with increased proliferation and can later be integrated into muscle fibers. Therefore, new myonuclei are available to enhance mRNA synthesis and protein expression in muscle cells. New understanding of the cellular mechanisms of signal transduction in muscle in response to training, bed rest and ageing will help to optimize training and interventions in an ageing population. PMID- 19308346 TI - [Effect of power training on articular cartilage]. AB - Hyaline articular cartilage consists mainly of water, which is reversibly transferred to the joint space under compression to reduce friction between the cartilage surfaces. This water content is regulated by proteoglycanes, which are produced by chondrocytes. Under excessive compression of the cartilage surface the collagen matrix ruptures and water is irreversibly transferred into the joint space, which reduces the elasticity of cartilage tissue. Power training should lead to a submaximal loading of cartilage below the rupture limit as the capacity of cartilage tissue to adapt to higher loads is limited. When the cartilage tissue shows osteoarthritic changes the loading capacity of the cartilage matrix is significantly reduced, which should be taken into consideration during power training. PMID- 19308347 TI - Polymorphic ROS scavenging revealed by CCCP in a lizard. AB - Ingestion of antioxidants has been argued to scavenge circulating reactive molecules (e.g., free radicals), play a part in mate choice (by mediating access to this important resource), and perhaps increase life span. However, recent work has come to question these relationships. We have shown elsewhere in the polychromatic lizard, Ctenophorus pictus, that diet supplementation of carotenoids as antioxidants does not depress circulating natural reactive oxygen species (ROS) levels and leads to no corresponding improvement of color traits. However, a much stronger test would be to experimentally manipulate the ROS levels themselves and assess carotenoid-induced ROS depression. Here, we achieve this by using carbonyl cyanide 3-chlorophenylhydrazone, which elevates superoxide (SO) formation approximately threefold at 10 microM in this model system. We then look for depressing effects on ROS of the carotenoids in order to assess whether 'super-production' of SO makes carotenoid effects on elevated ROS levels detectable. The rationale for this treatment was that if not even such elevated levels of SO are reduced by carotenoid supplementation, the putative link carotenoids, ROS depression, and mate quality (in terms of antioxidant capacity) is highly questionable. We conclude that there is no significant effect of carotenoids on mean SO levels even at the induced ROS levels. However, our results showed a significant interaction effect between carotenoid treatment and male color, with red males having higher ROS levels than yellow males. We suggest that this may be because different pigments are differently involved in the generation of the integumental colors in the two morphs with concomitant effects on ROS depletion depending on carotenoid uptake or allocation to coloration and antioxidation. PMID- 19308348 TI - No evidence for sperm priming responses under varying sperm competition risk or intensity in guppies. AB - Sperm competition theory predicts that males should tailor their investment in ejaculates according to the number of rival males competing to fertilize a female's eggs. Research spanning several taxa supports this prediction by showing that males are often sensitive to the level of sperm competition and adjust their investment in sperm numbers accordingly. More recent work has revealed that males may also tailor the quality of sperm according to the number of males competing for fertilization. Here I test for both effects in guppies (Poecilia reticulata) in an experiment that simultaneously evaluates the risk and intensity models of sperm competition. The experiment determined whether male guppies adjust the number (stripped ejaculate size) and quality (sperm velocity and viability) of sperm that are primed over a 3-day period according to experimental changes in the perceived level of sperm competition. A total of 136 focal males were initially stripped of all retrievable sperm and assayed for these sperm traits before being allocated at random to one of four treatments simulating different levels of sperm competition risk and intensity. During the 3-day treatment phase, focal males had visual and olfactory access to a sexually receptive (initially virgin) female maintained with different numbers of stimulus males to simulate variation in the risk and intensity of sperm competition. Following this, males were assayed again for the sperm traits. Contrary to predictions, there was no significant change in any of the measured variables among treatments, although qualitatively the patterns for sperm velocity and viability did conform to expectation. The lack of any trend for the number of sperm primed was unequivocal and future work examining the effects of sperm competition on sperm production should focus on whether males differentially allocate sperm numbers among matings that differ in the level of sperm competition. PMID- 19308349 TI - Paraquat resistance in a Lolium rigidum population is governed by one major nuclear gene. AB - Paraquat resistance in an annual ryegrass (Lolium rigidum Gaud.) population (AFLR1) has been attributed to reduced paraquat translocation. Genetic inheritance of paraquat resistance in this population was investigated in the present study. The paraquat dose response of progeny from 8 F(1) families was more similar to that of the resistant than the susceptible parent, while the equivalent data for a further three families were intermediate compared to those of the parental populations. No significant differences in dose response were observed between reciprocal crosses of specific F(1) families. These results suggest that paraquat resistance in AFLR1 is inherited as a dominant or partially dominant nuclear-encoded trait. Pseudo-F(2) (psi-F(2)) generation seedlings were treated with multiple dose rates sufficient to control the susceptible parental population, and observed segregation ratios in all instances conformed to a 3:1 (resistant:susceptible) segregation ratio, and this ratio was further confirmed by individual phenotyping of cloned plant genotypes. A single major nuclear gene is hence apparently responsible for evolved paraquat resistance in AFLR1. PMID- 19308350 TI - Variations in KCNQ1 are associated with type 2 diabetes and beta cell function in a Chinese population. AB - AIMS/HYPOTHESIS: Recent genome-wide association studies in East Asian populations reported that single nucleotide polymorphisms (SNPs) in KCNQ1 are associated with type 2 diabetes. The aim of this study was to validate this finding in a Chinese population. METHODS: We genotyped four SNPs, rs2074196, rs2237892, rs2237895 and rs2237897, in a group of 3,503 Shanghai Chinese individuals, comprising 1,769 type 2 diabetic patients and 1,734 normoglycaemic controls. Both the cases and the controls were extensively phenotyped for anthropometric and biochemical traits related to glucose metabolism. Arginine stimulation tests under fasting conditions were performed in a subgroup of 466 cases. RESULTS: All four of the SNPs were associated with type 2 diabetes, with rs2237892 showing strongest evidence for association (OR 1.532, 95% CI 1.381-1.698, p = 5.0 x 10(-16)). The SNP rs2237897 was associated with both acute insulin and C-peptide response after arginine stimulation in a subgroup of cases (p = 0.0471 and p = 0.0156, respectively). The SNP rs2237895 was associated with both first- and second-phase insulin secretion in the controls (p = 0.0334 and p = 0.0002, respectively). CONCLUSIONS/INTERPRETATION: In this study we found that KCNQ1 was associated with type 2 diabetes susceptibility in a Chinese population, possibly through its effect on beta cell function. PMID- 19308351 TI - The effect of bilio-pancreatic diversion on type 2 diabetes in patients with BMI <35 kg/m2. AB - AIMS/HYPOTHESIS: To aim of the study was to investigate the effect of bilio pancreatic diversion (BPD) on type 2 diabetes in patients with BMI <35 kg/m(2). METHODS: OGTTs were performed and anthropometric data were compared between five diabetes patients (BMI 27-33 kg/m(2)) following BPD and seven diabetes patients after a low-energy diet. Insulin secretion was computed by C-peptide deconvolution. A euglycaemic-hyperinsulinaemic clamp was performed only in the BPD group and the M value measured. RESULTS: One month after BPD, fasting and 2 h post-OGTT glycaemia decreased from 15.22 +/- 3.22 to 6.22 +/- 0.51 mmol/l (p = 0.043), while insulin sensitivity increased significantly. No significant changes were observed in the low-energy diet group. Insulin secretion did not differ significantly after either intervention. Diabetes amelioration (change in HbA(1c) level) was observed up to 18 months after BPD without pharmacological therapy. CONCLUSIONS/INTERPRETATION: BPD can achieve adequate control of type 2 diabetes also in patients with BMI <35 kg/m(2). The rapid postoperative remission of diabetes is primarily related to an improvement in insulin sensitivity. PMID- 19308352 TI - [Treatment of infected total knee arthroplasty. 2-5-year results following two stage reimplantation]. AB - BACKGROUND: The 2-5-year results for the treatment of deep infection of total knee arthroplasty (TKA) after two-stage reimplantation are presented. An articulating temporary antibiotic spacer prosthesis (TASP) and a standardized antibiotic regimen were used. PATIENTS AND METHODS: In a prospective study, 33 consecutive patients were treated with TASP. This articulating spacer was made on the table by cleaning and autoclaving removed parts of the TKA. Intravenous double antibiotic therapy in combination with rifampin was given for 10 days, followed by oral therapy for 4 weeks. RESULTS: At a mean follow-up period of 47 months (31-67), three patients had reinfection (success rate 91%). The average Hospital for Special Surgery knee score increased from 67 points (44-84) to 85 points (53-97) after reimplantation. Based on these results, 24 knees (73%) were rated excellent, five (15%) were rated good, three (9%) were rated fair, and one patient (3%) had a poor result. Complications included one case of temporary peroneal palsy, one dislocation of the spacer due to an insufficient extensor mechanism, and one fracture of the tibia due to substantial primary metaphyseal bone loss. CONCLUSION: Using TASP, the disadvantages of joint fixation between the two stages could be reduced. There was no difference in the reinfection rate compared with procedures using fixed spacer blocks. TASP facilitates reimplantation and yields good functional results. PMID- 19308353 TI - Sepsis care bundles and clinicians. PMID- 19308354 TI - Reduction of ventilator-associated pneumonia: active versus passive guideline implementation. AB - PURPOSE: Ventilator-associated pneumonia (VAP) is associated with increased morbidity, mortality and costs. We describe an active, multifaceted implementation of a VAP prevention bundle designed to improve staff compliance with evidence-based actions and reduce the incidence of VAP. METHOD: A 'VAP prevention bundle' was designed then implemented, first passively, then actively, as defined by a multimodal programme incorporating staff education, process measurement and outcome measurement and feedback to staff and organisational change. RESULTS: Compliance with the VAP prevention bundle increased after active implementation. VAP incidence fell significantly from 19.2 to 7.5 per 1,000 ventilator days. Rate difference (99% CI) = 11.6 (2.3-21.0) per 1,000 ventilator days; rate ratio (99% CI) = 0.39 (0.16, 0.96). CONCLUSIONS: An active implementation programme increased staff compliance with evidence-based interventions and was associated with a significant reduction in VAP acquisition. PMID- 19308355 TI - Hallux valgus in patients with patellofemoral pain syndrome. AB - The aim of this study was to investigate the presence of hallux valgus (HV) in patellofemoral pain syndrome (PFPS). Ninety-nine patients with unilateral patellofemoral pain participated. The study was designed with each subject acting as their own internal control by using the unaffected limb for comparison. The HV and intermetatarsal angles were measured by weight-bearing anteroposterior radiographs in patients. Eighty-four out of 99 patients had an abnormal HV angle on the affected side and 78 of 99 patients had an abnormal HV angle on the unaffected side. There were significant differences in the HV angle between affected and unaffected sides (P = 0.003). This study is the first to demonstrate the relationship between HV and PFPS. Further comprehensive biomechanical studies are warranted to analyse the relationship between HV and PFPS. PMID- 19308356 TI - Radiological evaluation of the anterolateral and posteromedial bundle insertion sites of the posterior cruciate ligament. AB - The optimal technique of posterior cruciate ligament (PCL) reconstruction is still controversial. Besides different parameters as surgical technique and graft choice, the exact anatomic placement of the graft is essential for successful reconstruction of the PCL. For intraoperative control and postoperative radiological evaluation of the anatomical placement of the anterolatelateral (ALB) and posteromedial bundle (PMB) of the PCL, the radiological localization of both functional bundles of the PCL has yet to be determined. Therefore, in this descriptive laboratory study, the insertion sites of the ALB and PMB of the PCL on the femur and the common tibial insertion site were macroscopically identified and marked with copper wires in 16 human cadaver specimens. Radiological evaluation of the femoral insertion sites of the ALB and PMB in AP full extension and true lateral radiography was performed using an angle segment transformation based on the tangent of the femoral condyles and a modified reticule system of the quadrant method, respectively. On the tibial site, the footprint of the PCL was defined as ratios of the geometric insertion site with respect to the mediolateral and sagittal diameter of the tibia in AP and true lateral radiography. Femorally, the geometric insertion points of the ALB and PMB were located on the reticule at x = 62 +/- 3%/y = 16 +/- 6% and x = 51 +/- 5%/y = 35 +/- 7%, respectively; the angle segments for the PMB were between 40 +/- 5 degrees and 56 +/- 6 degrees and for the ALB were between 56 +/- 6 degrees and 76 +/- 7 degrees. Tibially, the common insertion point of the PCL was located at 51 +/- 2% of the mediolateral diameter of the tibial plateau with respect to the lateral border and 13 +/- 2% inferiorly to the medial tibial plateau with respect to the sagittal diameter of the tibial plateau. In conclusion, the knowledge of the anatomical insertion sites of the PCL in standardized radiography may help the orthopaedic surgeon for correct intraoperative placement and postoperative evaluation of the tunnel placement. In addition, the data might be a useful tool for fluoroscopic-based navigation in PCL reconstruction. PMID- 19308357 TI - Phosphate accumulation of Acetobacter xylinum. AB - The cells of Acetobacter xylinum decreased phosphate concentration in the medium from 5 to 2.5 or 0.3 mM during incubation in the presence of Mg(2+) and glucose, or Mg(2+) and casamino acids, respectively. The prevalence of orthophosphate or polyphosphate in the biomass of A. xylinum depends on the medium composition. Under phosphate uptake in the presence of glucose, the content of orthophosphate in the biomass changed little, while that of polyphosphate increased fourfold. At incubation with casamino acids, the content of orthophosphate increased 15 times, while that of polyphosphate increased only 2.5 times. Some part of orthophosphate in this case seems to be bound with the cell surface. The polyphosphate chain length in the cells of A. xylinim increases under phosphate uptake. This increase is more noticeable in the presence of glucose. Casamino acids can be replaced by alpha-ketoglutaric acid in combination with (NH(4))(2)SO(4), or arginine, or glutamine, the catabolism of which results in formation of NH(4) (+) and alpha ketoglutarate. PMID- 19308358 TI - Differential roles of nitric oxide synthase isozymes in cardiotoxicity and mortality following chronic doxorubicin treatment in mice. AB - The roles of individual nitric oxide synthases (NOS) in anthracycline-related cardiotoxicity are not completely understood. We investigated the effects of a chronic treatment with doxorubicin (DOX) on knockouts of the individual NOS isozymes and on transgenic mice with myocardial overexpression of eNOS. Fractional shortening (FS) was reduced in untreated homozygous nNOS and iNOS knockouts as well as in eNOS transgenics. DOX-induced FS decrease in wild-type mice was attenuated only in eNOS knockouts, which were found to overexpress nNOS. No worsening of contractility was observed in DOX-treated eNOS transgenics and iNOS knockouts. Although the surviving DOX-treated nNOS knockouts exhibited no further impairment in contractility, most (70%) animals died within 7 weeks after treatment onset. In comparison to untreated wild-type hearts, the nitric oxide (NO) level was lower in hearts from DOX-treated wild-type mice and in all three untreated knockouts. DOX treatment had no effect on NO in the knockouts. These data indicate differential roles of the individual NOS in DOX-induced cardiotoxicity. Protection against DOX effects conferred by eNOS deletion may be mediated by a compensatory overexpression of nNOS. NOS inhibition-based prevention of anthracycline-induced cardiotoxicity should be eNOS-selective, simultaneously avoiding inhibiting nNOS. PMID- 19308359 TI - Multisyringe flow injection analysis hyphenated with liquid core waveguides for the development of cleaner spectroscopic analytical methods: improved determination of chloride in waters. AB - In this work, the hyphenation of the multisyringe flow injection analysis technique with a 100-cm-long pathlength liquid core waveguide has been accomplished. The Cl-/Hg(SCN)2/Fe3+ reaction system for the spectrophotometric determination of chloride (Cl(-)) in waters was used as chemical model. As a result, this classic analytical methodology has been improved, minimizing dramatically the consumption of reagents, in particular, that of the highly biotoxic chemical Hg(SCN)2. The proposed method features a linear dynamic range composed of two steps between (1) 0.2-2 and (2) 2-8 mg Cl- L(-1), thus extended applicability due to on-line sample dilution (up to 400 mg Cl- L(-1)). It also presents improved limits of detection and quantification of 0.06 and 0.20 mg Cl- L(-1), respectively. The coefficient of variation and the injection throughput were 1.3% (n = 10, 2 mg Cl- L(-1)) and 21 h(-1). Furthermore, a very low consumption of reagents per Cl- determination of 0.2 microg Hg(II) and 28 microg Fe3+ has been achieved. The method was successfully applied to the determination of Cl- in different types of water samples. Finally, the proposed system is critically compared from a green analytical chemistry point of view against other flow systems for the same purpose. PMID- 19308360 TI - Separation of malignant human breast cancer epithelial cells from healthy epithelial cells using an advanced dielectrophoresis-activated cell sorter (DACS). AB - In this paper, we successfully separated malignant human breast cancer epithelial cells (MCF 7) from healthy breast cells (MCF 10A) and analyzed the main parameters that influence the separation efficiency with an advanced dielectrophoresis (DEP)-activated cell sorter (DACS). Using the efficient DACS, the malignant cancer cells (MCF 7) were isolated successfully by noninvasive methods from normal cells with similar cell size distributions (MCF 10A), depending on differences between their material properties such as conductivity and permittivity, because our system was able to discern the subtle differences in the properties by generating continuously changed electrical field gradients. In order to evaluate the separation performance without considering size variations, the cells collected from each outlet were divided into size-dependent groups and counted statistically. Following that, the quantitative relative ratio of numbers between MCF 7 and MCF 10A cells in each size-dependent group separated by the DEP were compared according to applied frequencies in the range 48, 51, and 53 MHz with an applied amplitude of 8 V(pp). Finally, under the applied voltage of 48 MHz-8 V(pp) and a flow rate of 290 microm/s, MCF 7 and MCF 10A cells were separated with a maximum efficiency of 86.67% and 98.73% respectively. Therefore, our suggested system shows it can be used for detection and separation of cancerous epithelial cells from noncancerous cells in clinical applications. PMID- 19308361 TI - Commercialized rapid immunoanalytical tests for determination of allergenic food proteins: an overview. AB - Food allergies have become an important health issue especially in industrialized countries. Undeclared allergenic ingredients or the presence of "hidden" allergens because of contamination during the food production process pose great health risks to sensitised individuals. The EU directive for food labelling lists allergenic foods that have to be declared on food products by the manufacturers. The list includes gluten-containing cereals, crustaceans, eggs, fish, peanuts, soybeans, milk, various nuts (e.g. almond, hazelnut, and walnut, etc.), celery, mustard, sesame seeds, lupin, and molluscs. Reliable methods for detection and quantification of food allergens are needed that can be applied in a fast and easy-to-use manner, are portable, and need only limited technical equipment. This review focuses on the latest developments in food allergen analysis with special emphasis on fast immunoanalytical methods such as rapid enzyme-linked immunosorbent assays (ELISA), lateral-flow immunochromatographic assays (LFA) and dipstick tests. Emerging technologies such as immunochemical microarrays and biosensors are also discussed and their application to food allergen analysis is reviewed. Finally, a comprehensive overview of rapid immunochemical test kits that are currently available commercially is given in tabular form. PMID- 19308362 TI - Spatiotemporal architecture of cortical receptive fields and its impact on multisensory interactions. AB - Recent electrophysiology studies have suggested that neuronal responses to multisensory stimuli may possess a unique temporal signature. To evaluate this temporal dynamism, unisensory and multisensory spatiotemporal receptive fields (STRFs) of neurons in the cortex of the cat anterior ectosylvian sulcus were constructed. Analyses revealed that the multisensory STRFs of these neurons differed significantly from the component unisensory STRFs and their linear summation. Most notably, multisensory responses were found to have higher peak firing rates, shorter response latencies, and longer discharge durations. More importantly, multisensory STRFs were characterized by two distinct temporal phases of enhanced integration that reflected the shorter response latencies and longer discharge durations. These findings further our understanding of the temporal architecture of cortical multisensory processing, and thus provide important insights into the possible functional role(s) played by multisensory cortex in spatially directed perceptual processes. PMID- 19308363 TI - The effects of prolonged viewing of motion on short-latency ocular following responses. AB - The adaptive effects of prolonged viewing of conditioning motion on ocular following responses (OFRs) elicited by brief test motion of a random-dot pattern were studied in humans. We found that the OFRs were significantly reduced when the directions of the conditioning and test motions were the same. The effect of conditioning motion was still observed when the speeds of the conditioning and test motions did not match. The effect was larger when the conditioning duration was longer, and decayed over time with increased temporal separation between the conditioning and test periods. These results are consistent with the characteristics of motion adaptation on the initial smooth pursuit responses. We also obtained data suggesting that the persistence of the effect depends on visual stimulation in the time between the conditioning and test periods, and that the presence of a stationary visual stimulus facilitates recovery from the motion adaptation. PMID- 19308364 TI - Relation between postural control assessment with eyes open and centre of pressure visual feedback effects in healthy individuals. AB - Visual feedback (VFB) of the resultant centre of pressure (CP(Res)) is a potentially interesting technique for rehabilitation purposes. However, all past studies have not been unanimous in supporting its utility in a physical therapy program. The present study was therefore undertaken with the main aim of assessing whether insights from postural control without additional feedback could be gained to explain the possible VFB effects. The CP(Res) displacements of 65 healthy adults were analysed in two conditions (eyes-open and VFB) through various classical parameters (including surface and variances along each medio lateral and antero-posterior axes) and fractional Brownian motion (fBm) modelling. The results indicate that 69% of the sample were dependent on VFB, i.e. that the surface covered by the CP(Res) displacements was smaller during VFB than in the eyes-open condition. The comparison of the two subgroups (i.e. those who were dependant on VFB and those who were not) brought out significant differences in many classical and fBm parameters. In addition, the correlation between the degree of VFB dependency and the values measured during the eyes-open condition indicates that the more the subjects are VFB-dependent, the less they lean forward, the smaller the variance and the better the CP(Res) displacements are controlled over the longest time intervals. By specifying the links between the postural performance in an eyes-open standard condition and the degree of VFB dependency, a number of clues help identify the reasons for which some subjects do not succeed in using this technique, hence explaining the controversy about its use as a rehabilitation tool. PMID- 19308365 TI - O-demethylation of codeine to morphine inhibited by low-dose levomepromazine. AB - PURPOSE: Codeine/paracetamol (C/P) and levomepromazine (L) are frequently co administered for the treatment of acute back pain, but the efficacy/effectiveness of this combination drug therapy has not been evaluated. The demethylation of codeine to morphine is catalyzed by the polymorphic enzyme cytochrome P450 2D6 (CYP2D6), of which levomepromazine (methotrimeprazine) is a known inhibitor. The aim of this study was to investigate whether low-dose levomepromazine inhibits the formation of morphine from codeine in a patient population of homozygous extensive (EM) and heterozygous extensive (HEM) metabolizers of CYP2D6. METHODS: Our patient cohort consisted of 29 patients hospitalized for acute back pain who were randomized to a 24-h treatment with either C/P (60 mg codeine + 1000 mg paracetamol) four times daily or to L+C/P (levomepromazine 5 + 5 + 5 + 10 mg + C/P) four times daily. After zero-urine sampling (baseline), the treatment was started and urine collected for 24 h. Blood samples were later genotyped for the CYP2D6*3, *4, and *6 polymorphisms by the PCR (LightCycler system) and for the *5 polymorphism using long PCR, to identify EM and HEM and to eliminate CYP2D6 poor metabolizers. Urine samples were analyzed using the CEDIA immunoassay and gas chromatography-mass spectrometry after enzymatic hydrolysis of glucuronide conjugates. O-demethylation ratios of codeine were calculated as hydrolyzed (total) concentrations of morphine/morphine + codeine. RESULTS: Twenty-two of the patients fulfilled the inclusion criteria, of whom ten were EM (five C/P and five L+C/P) and twelve were HEM (six C/P and six L+C/P) for functional CYP2D6 alleles. In the EM group, the median O-demethylation ratio was significantly higher (P = 0.016, Mann-Whitney test) after the C/P treatment (0.092, range 0.041-0.096) than after the L+C/P treatment (0.031, range 0.009-0.042). However, there was no significant difference between these two treatments in either the HEM group [n = 12; 0.024 (range 0.011-0.042) vs. 0.026 (range 0.009-0.041), respectively; P = 1.00] or in the combined EM/HEM group [11 C/P + 11 L+C/P; 0.041 (range 0.011 0.096) vs. 0.030 (range 0.009-0.042), respectively; P = 0.122]. CONCLUSIONS: Our study revealed significant inhibition in the O-demethylation of codeine to morphine in homozygous EM of CYP2D6 treated with low-dose levomepromazine and codeine/paracetamol, compared to treatment with codeine/paracetamol only. No significant difference could be detected in HEM or in the mixed and heterogenous group of EM/HEM. In patients prescribed this drug combination, the amount of morphine generated by the O-demethylation of codeine may be insufficient for effective pain relief. The therapeutic effect of codeine in the treatment of acute back pain should be assessed with and without levomepromazine. PMID- 19308366 TI - Assessment of extracranial-intracranial bypass patency with 64-slice multidetector computerized tomography angiography. AB - INTRODUCTION: Extracranial-intracranial (EC/IC) bypass is a useful procedure for the treatment of cerebral vascular insufficiency or complex aneurysms. We explored the role of multidetector computed tomography angiography (MDCTA), instead of digital subtraction angiography (DSA), for the postoperative assessment of EC/IC bypass patency. METHODS: We retrospectively analyzed a consecutive series of 21 MDCTAs from 17 patients that underwent 25 direct or indirect EC/IC bypass procedures between April 2003 and November 2007. Conventional DSA was available for comparison in 13 cases. MDCTA used a 64-slice MDCT scanner (Aquilion 64, Toshiba). The proximal and distal patencies were analyzed independently on MDCTA and DSA by a neuroradiologist and a neurosurgeon. The bypass was considered patent when the entire donor vessel was opacified without discontinuity from proximal to distal ends and was visibly in contact with the recipient vessel. RESULTS: MDCTA depicted the patency status in every patient. Bypasses were patent in 22 cases, stenosed in one, and occluded in two. DSA always confirmed the results of the MDCTA (sensitivity = 100%, 95% CI = 0.655 1.0; specificity 100%, 95% CI = 0.05-1.0). CONCLUSIONS: MDCTA is a non-invasive and accurate exam to assess the postoperative EC/IC bypass patency and is a promising technique in routine follow-up. PMID- 19308367 TI - Development of an MRI rating scale for multiple brain regions: comparison with volumetrics and with voxel-based morphometry. AB - INTRODUCTION: We aimed to devise a rating method for key frontal and temporal brain regions validated against quantitative volumetric methods and applicable to a range of dementia syndromes. METHODS: Four standardised coronal MR images from 36 subjects encompassing controls and cases with Alzheimer's disease (AD) and frontotemporal dementia (FTD) were used. After initial pilot studies, 15 regions produced good intra- and inter-rater reliability. We then validated the ratings against manual volumetry and voxel-based morphometry (VBM) and compared ratings across the subject groups. RESULTS: Validation against both manual volumetry (for both frontal and temporal lobes), and against whole brain VBM, showed good correlation with visual ratings for the majority of the brain regions. Comparison of rating scores across disease groups showed involvement of the anterior fusiform gyrus, anterior hippocampus and temporal pole in semantic dementia, while anterior cingulate and orbitofrontal regions were involved in behavioural variant FTD. CONCLUSION: This simple visual rating can be used as an alternative to highly technical methods of quantification, and may be superior when dealing with single cases or small groups. PMID- 19308368 TI - Imaging findings in 512 children following all-terrain vehicle injuries. AB - BACKGROUND: Injuries related to all-terrain vehicle (ATV) use by children have increased in recent years, and the pattern of these injuries is not well known among radiologists. OBJECTIVE: Our purpose was to identify different radiologically diagnosed injuries in children suffering ATV-related trauma and determine associations among various injuries as well as between injuries and outcome. MATERIALS AND METHODS: The study included 512 consecutive children suffering from ATV injuries treated at a tertiary care pediatric hospital. All imaging studies were reviewed and correlated with injury frequency and outcome using multivariate analysis. RESULTS: Head injuries occurred in 244 children (48%) and in five of six deaths. Calvarial skull fractures occurred in 104 children and were associated with brain, subdural and epidural injuries. Brain and orbit injuries were associated with long-term disability. A total of 227 extremity fractures were present in 172 children (34%). The femur was the most commonly fractured bone. Nine children had partial foot amputations. Multiorgan injuries occurred in nearly half of the 97 children with torso injuries. Determinants for long-term disability or death were head injuries (odds ratio 3.4) and extremity fractures (odds ratio 3.3). CONCLUSION: Head and extremity injuries are the two most common injuries in children suffering ATV injuries and are associated with long-term disability. ATV use by children is dangerous and is a significant threat to child safety. PMID- 19308372 TI - Pyloric stenosis: role of imaging. PMID- 19308373 TI - Intussusception in children: evidence-based diagnosis and treatment. PMID- 19308374 TI - Optimizing the role of imaging in appendicitis. AB - Acute appendicitis is the most common acute abdominal condition that requires surgical intervention in childhood. From the diagnostic performance perspective, computed tomography (CT) has a significantly higher sensitivity than does ultrasound (US) for diagnosing appendicitis in children; from the safety perspective, however, one should consider the radiation associated with CT, especially in children. There is strong evidence supporting improved patient outcomes in children with suspected acute appendicitis who undergo CT scanning. Nevertheless, we should keep in mind that for a single abdominal CT study in a 5 year-old child, the lifetime risk of radiation-induced cancer would be 26.1 per 100,000 in female and 20.4 per 100,000 in male patients, based on probabilistic models designed with data from atomic bomb survivors. An integrated clinical imaging approach, applying clinical scores that are able to predict which children with acute abdominal pain do or do not have a high probability of presenting with appendicitis may improve the effectiveness of the imaging diagnosis of appendicitis at the hospital level. Such an approach could avoid exposure of children who at low risk for appendicitis to unnecessary diagnostic tests and eventually, to radiation. PMID- 19308375 TI - Imaging choices in inflammatory bowel disease. PMID- 19308376 TI - Pancreatitis and the role of US, MRCP and ERCP. AB - Imaging plays a crucial role in the diagnosis of acute and chronic pancreatitis in children. Ultrasound (US) is the primary imaging modality. The US study can be improved by incorporating high resolution imaging, color Doppler, harmonic imaging and panorama view. Computer tomography (CT) is widely used for further evaluation. MR imaging in combination with MR cholangiopancreaticography (MRCP) is emerging as the modality of choice. It is non-invasive and radiation-free. It has high potential to replace endoscopic retrograde cholangiopancreaticography (ERCP), too. The latter is becoming more of an interventional tool. This review discusses the current status and comparative diagnostic potential of US, MRCP and ERCP. PMID- 19308377 TI - Abdominal trauma imaging: imaging choices and appropriateness. PMID- 19308378 TI - Evidence-based diagnosis of malrotation and volvulus. PMID- 19308379 TI - Malrotation: the balance of evidence. PMID- 19308380 TI - Surgical concerns in malrotation and midgut volvulus. AB - Appropriate management of the child with malrotation and/or midgut volvulus requires a multi-disciplinary approach with early clinical suspicion, rapid confirmation of diagnosis, and expedient operative intervention in order to obtain the most effective outcomes. PMID- 19308381 TI - The position of the duodenojejunal junction: the wrong horse to bet on in diagnosing or excluding malrotation. AB - PURPOSE: The purpose of this communication is to highlight the shortcomings of all currently used imaging criteria in diagnosing or excluding malrotation and offer ultrasound demonstration of the 3(rd) portion of the duodenum (D3) between the AO and the SMA in transverse and sagittal plains as the most reliable diagnostic method. BACKGROUND: Although UGI is currently considered to be the imaging modality of choice in diagnosis of malrotations, numerous publications indicate that in certain patients, false positives and negatives can be encountered. MATERIALS AND METHODS: The material consists of more than 10 years experience in university settings, during which the author has used US as the definitive imaging modality for the work-up of malrotation. High resolution linear transducers (5-17 MHz) are the transducers of choice. Imaging plains: A. Transverse. With gradual grading compression, the following landmarks are illustrated in cehaplocaudad directions in the following order. The junction of splenic vein with the SMV portal vein. The cross sections of SMA and SMV that may either be situated in midline, or to the right or the left of the midline. Left renal vein crossing the spine from left to right between the AO and the SMA. The jejunal vein, often coming from left, transversing between the AO and the SMA. The transverse portion of the duodenum, D3, between the AO & the SMA. B. Sagittal. D3 between vertically oriented SMA-SMV and the AO. If SMA is not aligned with the AO by slight compression on the right or the left side of the abdomen, it will be aligned (depending on leftward or rightward position of SMA SMV in axial plain). Vertical orientation of SMA and SMV if they have an anteroposterior orientation. C. Coronal. Side by side orientation of SMA and SMV if they don't have an anteroposterior orientation. RESULTS: In overwhelming majority of cases, by illustrating a retromesenteric D3 malrotation and, therefore, midgut volvulus were excluded. DISCUSSION: None of the current imaging criteria addresses the following most fundamental anatomic and embryologic facts regarding the gut rotation and fixation. In first trimester, the D3 is secured in retroperitoneal space after the embryologic journey comes to an end, making the duodenum immune of midgut volvulus. The surgical pathology of malrotation-midgut volvulus indicates that D3 is always Intraperitoneal and has not reached its final embryologic destination in retroperitoneal space. Demonstrating a retromesenteric D3, therefore, indicates that the embryologic journey is completed and the patient does not have malrotation. Excluding malrotation excludes the likelihood of midgut volvus. CONCLUSION: The position of the DJJ, the configuration of the duodenal sweep, the orientation of the mesenteric vessels are all wrong horses to bet on because none of them addresses the fundamental anatomic and embryologic facts. Only the cross-sectional imaging, US, CT and MRI can prove that the D3 is retromesenteric, therefore, excluding malrotation and volvulus. Therefore, demonstrating a retromesenteric duodenum is the reference standard of imaging in the work-up of malrotation, not any other previously published criteria. The US imaging is the most acceptable imaging method for malrotation work-up, in the spirit of ALARA principle and "Image Gently" campaign. PMID- 19308382 TI - What imaging should we perform for the diagnosis and management of pulmonary infections? PMID- 19308383 TI - Evaluation of non-vascular mediastinal masses in infants and children: an evidence-based practical approach. PMID- 19308384 TI - What is the optimal imaging for vascular rings and slings? PMID- 19308385 TI - The role of MRI and CT in congenital heart disease. PMID- 19308386 TI - What is the evidence based role of US in evaluating the fetus? PMID- 19308387 TI - The growing role of MR imaging in the fetus. PMID- 19308388 TI - The role of radiographs and US in developmental dysplasia of the hip: how good are they? PMID- 19308389 TI - What is the optimal imaging of osteonecrosis, Perthes, and bone infarcts? PMID- 19308390 TI - Impact of obesity in the diagnosis of SCFE and knee problems in obese children. PMID- 19308391 TI - How should we image skeletal injuries in child abuse? PMID- 19308392 TI - Imaging of nonaccidental head injury. PMID- 19308393 TI - Imaging strategies for new onset seizures. PMID- 19308394 TI - Diagnostic imaging in 2009: update on evidence-based practice of pediatric imaging. What is the role of imaging in sinusitis? PMID- 19308395 TI - Spinal dysraphism: categorizing risk to optimize imaging. PMID- 19308396 TI - Role of imaging in scoliosis. PMID- 19308400 TI - Hydrodynamic multibead modeling: problems, pitfalls, and solutions. 2. Proteins. AB - Hydrodynamic models of proteins have been generated by recourse to crystallographic data and applying a filling model strategy in order to predict both hydrodynamic and scattering parameters. The design of accurate protein models retaining the majority of the molecule peculiarities requires usage of many beads and consideration of many serious problems. Applying the expertise obtained with ellipsoid models and pilot tests on proteins, we succeeded in constructing precise models for several anhydrous and hydrated proteins of different shape, size, and complexity. The models constructed consist of many beads (up to about 11,000) for the protein constituents (atoms, amino acid residues, groups) and preferentially bound water molecules. While in the case of small proteins, parameter predictions are straightforward, computations for giant proteins necessitate drastic reductions of the number of initially available beads. Among several auxiliary programs, our advanced hydration programs, HYDCRYST and HYDMODEL, and modified versions of Garcia de la Torre's program HYDRO were successfully employed. This allowed the generation of realistic protein models by imaging details of their fine structure and enabled the prediction of reliable molecular parameters including intrinsic viscosities. The appearance of the models and the agreement of molecular properties and distance distribution functions p(r) of unreduced and reduced models can be used for a meticulous inspection of the data obtained. PMID- 19308401 TI - Chemical characterization of soil extract as growth media for the ecophysiological study of bacteria. AB - We investigated the composition of soil-extracted solubilized organic and inorganic matter (SESOM) prepared from three different soils. Growth of various bacterial strains in these soil extracts was evaluated to find appropriate conditions for ecophysiological approaches. Analysis of SESOM by (1)H-NMR and gas chromatography/mass spectrometry revealed a complex mixture of organic compounds. An oak forest SESOM supported the growth of several gram-positive and gram negative soil-derived heterotrophic bacteria, whereas beech forest and grassland soil extracts did not. A metabolomic approach was performed by determining the extracellular metabolite profile of Bacillus licheniformis in SESOM. The results demonstrated that determination of the organic composition of SESOM during batch culturing is feasible. This makes SESOM amenable to studying the ecophysiology of a range of soil bacteria growing on soil-dissolved organic matter under more defined laboratory conditions. SESOM may also increase success in isolating previously uncultured or novel soil bacteria. Cell populations and the corresponding extracellular medium can be obtained readily and specific components extracted, paving the way for proteomic, transcriptomic, and metabolomic analyses. The synthetic carbon mixture based on SESOM, which mimics soil abilities, shows a positive impact on higher cell yields and longer cultivation time for biotechnological relevant bacteria. PMID- 19308402 TI - A recombinant E. coli bioprocess for hyaluronan synthesis. AB - An Escherichia coli strain, JM109, was successfully engineered into an efficient hyaluronic acid (HA) producer by co-expressing the only known class-II HA synthase from a Gram-negative bacterium (Pasteurella multocida) and uridine diphosphate-glucose dehydrogenase from E. coli K5 strain. The engineered strain produced about 0.5 g/L HA in shake flask culture and about 2.0-3.8 g/L in a fed batch fermentation process in a 1-L bioreactor. The sharp increase in viscosity associated with HA accumulation necessitated pure oxygen supplement to maintain fermentation in aerobic regime. Precursor supply during HA synthesis was probed by glucosamine supplement, which shortens biosynthesis pathway and eliminates one step requiring ATP. HA synthesis was increased with glucosamine supplement from 2.7 to 3.7 g/L (37%), which was mirrored with a concomitant 42% decrease in pure oxygen input, suggesting a close connection between energy metabolism and precursor supply. Decoupling HA synthesis from cell growth by using fosfomycin (an inhibitor for cell wall synthesis) led to a 70% increase in HA synthesis, suggesting detrimental effects on HA synthesis from cell growth via precursor competition. This study demonstrates a potentially viable process for HA based on a recombinant E. coli strain. In addition, the precursor supply limitation identified in this study suggests new engineering targets in subsequent metabolic engineering efforts. PMID- 19308403 TI - Biosynthesis and biotechnological production of serotonin derivatives. AB - Serotonin derivatives belong to a class of phenylpropanoid amides found at low levels in a wide range of plant species. Representative serotonin derivatives include feruloylserotonin (FS) and 4-coumaroylserotonin (CS). Since the first identification of serotonin derivatives in safflower seeds, their occurrence, biological significance, and pharmacological properties have been reported. Recently, serotonin N-hydroxycinnamoyl transferase (SHT), which is responsible for the synthesis of serotonin derivatives, was cloned from pepper (Capsicum annuum) and characterized in terms of its enzyme kinetics. Using the SHT gene, many attempts have been made to either increase the level of serotonin derivatives in transgenic plants or produce serotonin derivatives de novo in microbes by dual expression of key genes such as SHT and 4-coumarate-CoA ligase (4CL). Due to the strong antioxidant activity and other therapeutic properties of serotonin derivatives, these compounds may have high potential in treatment and prophylaxis, as cosmetic ingredients, and as major components of functional foods or feeds that have health-improving effects. This review examines the biosynthesis of serotonin derivatives, corresponding enzymes, heterologous production in plants or microbes, and their applications. PMID- 19308404 TI - A T7 RNA polymerase-dependent gene expression system for Bacillus megaterium. AB - Gene expression systems based on the RNA polymerase of the bacteriophage T7 are often the ultimate choice for the high level production of recombinant proteins. During the last decade, the Gram-positive bacterium Bacillus megaterium was established as a useful host for the intra- and extracellular production of heterologous proteins. In this paper, we report on the development of a T7 RNA polymerase-dependent expression system for B. megaterium. The system was evaluated for cytosolic and secretory protein production with green fluorescent protein (GFP) from Aequoria victoria as intracellular and Lactobacillus reuteri levansucrase as extracellular model protein. GFP accumulated rapidly at high levels up to 50 mg/l shake flask culture intracellularly after induction of T7 RNA polymerase gene expression. The addition of rifampicin for the inhibition of B. megaterium RNA polymerase led to an increased stability of GFP. L. reuteri levansucrase was also successfully produced and secreted (up to 20 U/l) into the culture supernatant. However, parallel intracellular accumulation of the protein indicated limitations affiliated with the Sec-dependent protein translocation process. PMID- 19308405 TI - In vitro model to study the modulation of the mucin-adhered bacterial community. AB - There is an upsurge of interest in gastro-intestinal microbiology to improve the balance between positive and negative commensals. Mucosal bacteria make closer contact with the host than luminal ones and can therefore have a stronger health impact. An in vitro adhesion assay was developed to study the mucin colonization of bacteria from the mixed microbial communities of the Simulator of the Human Intestinal Microbial Ecosystem. Adhesion capacity differed substantially between bacteria and decreased from lactobacilli over fecal coliforms, bifidobacteria, and clostridia to total anaerobes. Lactobacillus rhamnosus GG adhered most selectively. Further, intestinal water lowered adhesion compared to phosphate buffered saline. By processing the data to an Adhesion-Related Prebiotic Index, it was found that intestinal water stimulated adherence of positive commensals. Arabinoxylans decreased the adhesion capacity matrix independently, whereas inulin had less or no influence. Measurements of bacterial surface tension, surface hydrophobicity, liquid surface tension, and viscosity showed that bacterial adhesion to mucin agar is a matter of both non-specific and specific interactions. The developed methodology can be useful for the characterization of the relevant but barely investigated mucin-associated bacterial community in health and disease (e.g., IBD) as well as for its modulation with functional foods like prebiotics. PMID- 19308406 TI - Variations in normal sternoclavicular joints; a retrospective study to quantify SCJ asymmetry. AB - OBJECTIVE: A wide degree of normal anatomical variation can occur at the sternoclavicular joint (SCJ). On occasion, this has led to concern for a pathological process, potentially resulting in a costly work-up, unnecessary patient worry and invasive diagnostic procedures such as biopsy. The purpose of this study was to determine the normal range of anatomical variation at sternoclavicular joints. MATERIALS AND METHODS: One hundred four consecutive patients with chest CT done at our institution were selected. Patients with clear SCJ pathology, chest wall abnormality, CT slice thickness greater than 5 mm and sternotomy wires, were excluded. Chart review was done and showed no SCJ symptoms/signs. We measured the SCJ space, maximum clavicular head diameter within the joint and the distance from manubrium to the anterior margin of the clavicular head. RESULTS: Left and right SCJ space ranged from 0.2 to 1.37 cm. The difference (delta or asymmetry) between left SCJ space and right SCJ space ranged from 0 (symmetrical) to 0.57 cm in 104 cases. Left and right clavicular head diameter ranged from 1.2 to 3.7 cm with left/right asymmetry (delta) ranging from 0 (symmetrical) to 1 cm. Manubrium to anterior margin of clavicular head ranged from 0.1 to 2.13 cm with delta ranging from 0 to 0.8 cm. Thirty-three patients demonstrated gas in the joint, five had poor articulation and four had calcification in the joint. CONCLUSION: Greater than 10% of patients show substantial asymmetry in the sternoclavicular joints, which may be misinterpreted as pathological. Gas in the joint is a common phenomenon therefore should not be an indication for further work-up in asymptomatic patients and likely excludes the presence of effusion. PMID- 19308407 TI - Free vascularised fibular grafting in the treatment of large skeletal defects due to osteomyelitis. AB - Treatment of skeletal defects secondary to osteomyelitis is a challenging problem. The purpose of this study was to present our experience of the use of free vascularised fibular grafts to treat such defects. Ten patients with a mean age of 31 years (range 16-50 years) and a skeletal defect with a mean length of 9.5 cm (range 6-17 cm) were managed with a protocol which included radical debridement of the lesion and a vascularised fibular graft. The mean follow-up time was 26 months. Union of the graft occurred in all patients, at a mean of 4.5 months. No recurrence of osteomyelitis was observed. The mean time to full weight bearing was ten months, and all patients were pain-free and able to walk without supportive devices. A free vascularised fibular graft is a viable option for the management of large skeletal defects resulting from osteomyelitis. PMID- 19308408 TI - Human cartilage glycoprotein 39--biomarker of joint damage in knee osteoarthritis. AB - Human glycoprotein of cartilage (YKL-40) synthesizes chondrocytes and synovial cells in inflammatory conditions or remodels the outer cell matrix in osteoarthritis. The aim of this study was to conduct a parallel analysis between thickness of cartilage and length of osteophytes, ultrasound indicators of joint destruction, with levels of YKL-40 in serum in patients with primary osteoarthritis. Ultrasound findings and concentration of YKL-40(ng/ml) were examined in 88 patients. The average value cartilage thickness measured on medial condyles of the femur was 1.30 +/- 0.23 mm and on lateral was 1.39 +/- 0.27 mm. Median YKL-40 in patients with shorter osteophytes was 62.0 (44.5-90) ng/ml, and with longer osteophytes was 119 (range 80-171) ng/ml (p = 0.000). YKL-40 can be a marker for the appearance of longer osteophytes (sensitivity = 79.1%; specificity = 61.9%; cut off = 75.0 ng/ml). The duration of illness is very much connected to values of YKL-40 (r = 0.651, p = 0.000). After an illness duration of five years, the concentration of YKL-40 was 83.68 +/- 33.65 ng/ml, after ten years it was 138.22 +/- 48.88 ng/ml, and after 15 and 20 years it was 209.30 +/- 79.36 ng/ml and 218.50 +/- 106.51 ng/ml, respectively. Higher concentrations of YKL-40 indicate the level of cartilage destruction and can be used for assessment of destruction. PMID- 19308409 TI - Thalidomide suppressed the growth of 4T1 cells into solid tumors in Balb/c mice in a combination therapy with the oncolytic fusogenic HSV-1 OncdSyn. AB - PURPOSE: The anti-tumor properties of thalidomide or in combination with an oncolytic herpes virus (OncdSyn) was investigated in a mouse model of human breast cancer. METHODS: To determine if thalidomide could act alone, 4T1 cells were injected into Balb/c mice. Tumors were sized, and the mice were fed chow or chow-containing thalidomide. After 4 days the tumor volumes were compared. To determine if thalidomide could act with the virus, tumors of mice were injected with phosphate buffered saline (PBS), or fed thalidomide with injections of PBS, or fed thalidomide with injections of OncdSyn, or received injections of OncdSyn. RESULTS: Thalidomide alone suppressed tumor growth. The most significant treatment occurred in thalidomide-fed-OncdSyn-injected mice. Compared to PBS controls, there was a significant difference in the number of metastatic nodes in the lungs. CONCLUSIONS: Thalidomide alone delayed tumor growth, but the combination of thalidomide with OncdSyn appeared to produce the best results. PMID- 19308410 TI - Phase I, dose escalation and pharmacokinetic study of cediranib (RECENTIN), a highly potent and selective VEGFR signaling inhibitor, in Japanese patients with advanced solid tumors. AB - PURPOSE: To evaluate safety and tolerability of cediranib, a highly potent and selective vascular endothelial growth factor signaling inhibitor, in Japanese patients with advanced solid tumors refractory to standard therapies. METHODS: In part A (n = 16), patients received once-daily oral cediranib (10-45 mg) to identify the maximum tolerated dose (MTD). In part B (n = 24), patients with non small-cell lung cancer or colorectal cancer received multiple daily doses at the MTD. RESULTS: Cediranib 30 mg/day was considered the MTD since 50% of evaluable patients receiving 45 mg/day experienced dose-limiting toxicities in part A (proteinuria and diarrhea n = 1, proteinuria n = 1, thrombocytopenia n = 1). The most common adverse events were diarrhea (n = 34) and hypertension (n = 32). Pharmacokinetic analysis confirmed cediranib as suitable for once-daily oral dosing. Of 32 evaluable patients, two had partial RECIST responses and 24 had stable disease > or =8 weeks. CONCLUSIONS: Cediranib was generally well tolerated at < or =30 mg/day in these Japanese patients and showed encouraging antitumor activity. PMID- 19308411 TI - Antineoplastic activity of idazoxan hydrochloride. AB - PURPOSE: Idazoxan hydrochloride (IDA) is a 241 molecular weight imidazoline and adrenoreceptor ligand. It binds to mitochondrial membranes and promotes apoptosis of pancreatic beta cells. Since IDA has not been tested against tumor cells, the purpose of our study was to determine if IDA has antineoplastic activity. METHODS: We used the conversion of a soluble tetrazolium salt to an insoluble formazan precipitate and differential staining cytotoxicity assays to determine if IDA was cytotoxic to cell lines of murine lung cancer and human prostate cancer, as well as to a variety of fresh human tumor samples. We used flow cytometry to analyze cell death and calreticulin expression. RESULTS: IDA is cytotoxic to both cell lines and against aliquots of specimens of breast, gastric, lung, ovarian and prostate cancers as well as non-Hodgkin's lymphoma. It produces apoptotic cell death and promotes calreticulin expression, suggesting that IDA might be immunomodulatory in vivo. CONCLUSION: We anticipate that IDA will be clinically useful in cancer treatment. PMID- 19308412 TI - Overcoming resistance to bisphosphonates through the administration of alfacalcidol: results of a 1-year, open follow-up study. AB - This study intended to determine whether the replacement of vitamin D3 with alfacalcidol results in any bone mineral density (BMD) increase in 76 patients unresponsive to the combination of alendronate and conventional vitamin D3 treatment. In these patients the conventional vitamin D3 had been replaced with alfacalcidol (0.5 MUg/day), and then the patients were followed up for a year. After treatment for 1 year, Wilcoxon test revealed a small but statistically significant (P < 0.001) increase in the BMD values of the forearm and lumbar vertebrae, in the serum calcium and urinary calcium/creatinine ratio in first voided morning urine. However, the serum alkaline phosphatase activity, phosphorus, parathormone, osteocalcin levels and the urinary d-pyr/creatinine ratio decreased significantly (P < 0.001). As suggested by our results, combination therapy with alendronate and alfacalcidol increases bone density and improves the biochemical markers of bone turnover, without any substantial increase in the incidence of adverse effects. PMID- 19308413 TI - Evidence for stable transformation of wheat by floral dip in Agrobacterium tumefaciens. AB - Hexaploid wheat, one of the world's most important staple crops, remains a challenge for genetic transformation. We are developing a floral transformation protocol for wheat that does not require tissue culture. This paper presents three transformants in the hard red germplasm line Crocus that have been characterized thoroughly at the molecular level over three to six generations. Wheat spikes at the early boot stage, i.e. the early, mid or late uninucleate microspore stages, were immersed in an infiltration medium of strain C58C1 harboring pDs(Hyg)35S, or strain AGL1 harboring pBECKSred. pDs(Hyg)35S contains the NPTII and hph selectable markers, and transformants were detected using paromomycin spray at the whole plant level, NPTII ELISAs, or selection on medium with hygromycin. Strain AGL1, harboring pBECKSred, which contains the maize anthocyanin regulators, Lc and C1, and the NPTII gene, was also used to produce a Crocus transformant. T1 and T2 seeds with red embryos were selected; T1 and T2 plants were screened by sequential tests for paromomycin resistance and NPTII ELISAs. The transformants were low copy number and showed Mendelian segregation in the T2. Stable transmission of the transgenes over several generations has been demonstrated using Southern analysis. Gene expression in advanced progeny was shown using Reverse Transcriptase-PCR and ELISA assays for NPTII protein expression. This protocol has the potential to reduce the time and expense required for wheat transformation. PMID- 19308414 TI - Diffusion-weighted magnetic resonance imaging of pancreas tumours. AB - The purpose of this study was to evaluate the accuracy of diffusion-weighted imaging (DWI) in diagnosis of pancreas cancer, to compare DWI with a conventional comprehensive MRI (MRI-c) and to analyse apparent diffusion coefficient (ADC) values of lesions. Thirty-six patients with pancreatic lesions (12 malignant and 24 benign) and 39 patients without lesions were included. MRI-c and DWI (free breathing, b values 0 and 500 s/mm(2)) were performed prospectively and consecutively in a 1.5-T system. The analysis was retrospectively performed blinded by two radiologists in consensus. The sensitivity, specificity, accuracy, and positive and negative predictive values of DWI and MRI-c were 92, 97, 96, 85, 98% and 100, 97, 97, 86, 100%, respectively. Mean ADC values of malignant lesions were significantly lower than those of benign lesions. DWI has a similar accuracy to MRI-c in diagnosis of pancreas cancer. PMID- 19308415 TI - Functional anatomy of the dorsal hood or the hand: correlation of ultrasound and MR findings with cadaveric dissection. AB - The main aim of this study was to provide an overview of the anatomy of the dorsal hood (DH) based on the dissection of sixteen cadaver hands, correlated with magnetic resonance (MR) and ultrasound findings. A secondary aim was to assess the function of components of the DH. Sixteen embalmed hands were evaluated by MRI and ultrasound before being dissected. Each hand was photographed during each stage of dissection. Secondly we evaluated the role of the different structures of the DH in the stability of the extensor tendon by transection of the different components alternatively at the ulnar and radial sides. MR, ultrasound, and dissection showed that the extensor tendon (ET) is stabilized by the sagittal band (SB) at the level of the metacarpophalangeal (MCP) joint and more distally by the transverse and the oblique bands, respectively. Transection of the radial SB of the second finger leads systematically to ulnar dislocation of the ET. The transection of the ulnar DH does not lead to instability of the ET. The SB is the most important structure of the DH in the stability of the ET at the MCP level. Rupture of the radial SB of the second finger leads systematically to ulnar dislocation of the ET. PMID- 19308416 TI - MRI of small bowel Crohn's disease: determining the reproducibility of bowel wall gadolinium enhancement measurements. AB - This study aims to determine inter- and intra-observer variation in MRI measurements of relative bowel wall signal intensity (SI) in Crohn's disease. Twenty-one small bowel MRI examinations (11 male, mean age 40), including T1 weighted acquisitions acquired 30 to 120s post-gadolinium, were analysed. Maximal bowel wall SI (most avid, conspicuous contrast enhancement) in designated diseased segments was measured by two radiologists and two trainees using self positioned "free" regions of interest (ROIs) followed by "fixed" ROIs chosen by one radiologist, and this procedure was repeated 1 month later. Relative enhancement (post-contrast SI minus pre-contrast SI/pre-contrast SI) was calculated. Data were analysed using Bland-Altman limits of agreement and intra class correlation. Inter-observer agreement for relative enhancement was poor (spanning over 120%) using a free ROI-95% limits of agreement -0.69, 0.70 and 0.47, 0.74 for radiologists and trainees, respectively, only marginally improved by use of a fixed ROI -0.60, 0.67 and -0.59, 0.49. Intra-class correlation ranged from 0.46 to 0.72. Intra-observer agreement was slightly better and optimised using a fixed ROI-95% limits of agreement -0.52, 0.50 and -0.34, 0.28 for radiologists and trainees, respectively. Intra-class correlation ranged from 0.49 to 0.86. Relative bowel wall signal intensity measurements demonstrate wide limits of observer agreement, unrelated to reader experience but improved using fixed ROIs. PMID- 19308417 TI - MR spectroscopy (MRS) and magnetisation transfer imaging (MTI), lesion load and clinical scores in early relapsing remitting multiple sclerosis: a combined cross sectional and longitudinal study. AB - The purpose of this study was to correlate magnetic resonance imaging (MRI)-based lesion load assessment with clinical disability in early relapsing remitting multiple sclerosis (RRMS). Seventeen untreated patients (ten women, seven men; mean age 33.0 +/- 7.9 years) with the initial diagnosis of RRMS were included for cross-sectional as well as longitudinal (24 months) clinical and MRI-based assessment in comparison with age-matched healthy controls. Conventional MR sequences, MR spectroscopy (MRS) and magnetisation transfer imaging (MTI) were performed at 1.5 T. Lesion number and volume, MRS and MTI measurements for lesions and normal appearing white matter (NAWM) were correlated to clinical scores [Expanded Disability Status Scale (EDSS), Multiple Sclerosis Functional Composite (MSFC)] for monitoring disease course after treatment initiation (interferon beta-1a). MTI and MRS detected changes [magnetisation transfer ratio (MTR), N-acetylaspartate (NAA)/creatine ratio] in NAWM over time. EDSS and lesional MTR increases correlated throughout the disease course. Average MTR of NAWM raised during the study (p < 0.05) and correlated to the MSFC score (r = 0.476, p < 0.001). At study termination, NAA/creatine ratio of NAWM correlated to the MSFC score (p < 0.05). MTI and MRS were useful for initial disease assessment in NAWM. MTI and MRS correlated with clinical scores, indicating potential for monitoring the disease course and gaining new insights into treatment-related effects. PMID- 19308418 TI - [Indications and technique of optic nerve sheath fenestration]. AB - An optic nerve sheath fenestration is indicated when papilloedema leads to progressive visual loss despite previous, conservative measures and when no cause of increased intracranial pressure can be indentified and eliminated. This rather rare constellation usually occurs in idiopathic intracranial hypertension. The procedure is performed via a medial transconjunctival orbitotomy. If headaches and neurologic symptoms other than visual deterioration prevail, the placement of a ventricular shunt is preferred. This review covers the symptoms of idiopathic intracranial hypertension and optic nerve sheath fenestration with its technical aspects and results. PMID- 19308419 TI - [Influence of antiangiogenetic therapy on retinal thickness values in age-related macular degeneration]. AB - BACKGROUND: This study evaluated the impact of antiangiogenetic therapy on differences in central retinal thickness (CRT) as measured with two optical coherence tomography (OCT) systems from the same manufacturer. MATERIAL AND METHODS: Data were obtained from a group of 20 patients with neovascular age related macular degeneration before and after intravitreal treatment with ranibizumab. Imaging was performed using the Stratus and the high-definition (HD) Cirrus OCT systems. RESULTS: The mean CRT was revealed to be significantly lower as measured by OCT compared with HD-OCT before and after treatment (p<0.002). CRT differences varied more strongly before treatment than after treatment. A higher standard deviation was noted before treatment. The difference in the mean deviation of CRT measurements before and following antiangiogenetic therapy was highly significant. CONCLUSION: This study showed that differences in CRT measurements between different OCT systems were subject not only to technical differences but also to treatment effects of current antiangiogenetic strategies. These effects should be recognized because clinical studies increasingly define OCT parameters as primary or secondary outcome measures. PMID- 19308420 TI - [Computer animated childrens pictures for vision testing]. AB - BACKGROUND: Testing visual acuity is a very important element especially in the ophthalmological examination of children. The diagnosis and procedure of subsequent therapy depend on the results of vision testing. Some children achieve false low results in visual acuity because of reduced compliance. This can lead to incorrect diagnoses and false therapies. PATIENTS AND METHODS: We developed a computer animated version of the visual acuity chart "Echelle Animaux". This new test was first performed on 16 normal adult volunteers and then on 16 children who achieved as good visual scores in conventional tests as the control group. After that we tested 25 children with low compliance. RESULTS: Most of the tested children were more motivated and attentive when performing the new test. In 9 out of 25 children the measured visual acuity scores were clearly better than in the conventional test. CONCLUSION: The animation of optotypes has a favourable effect on the motivation of children. The problem of reduced compliance and concentration can be reduced. The visual acuity scores can be improved by eliminating mistakes caused by low motivation. PMID- 19308421 TI - Induced preference for host plant chemicals in the tobacco hornworm: contribution of olfaction and taste. AB - Many herbivorous insects induce preferences for host plants. Recent work in Manduca sexta indicates that induced preferences are mediated by a "tuning" of the peripheral taste system to chemicals within host plant foliage. We tested this hypothesis by rearing caterpillars on artificial diet or potato foliage, and then examining olfactory- and taste-mediated responses to potato foliage extract. First, we confirmed earlier reports that consumption of potato foliage tunes the peripheral taste system by reducing responsiveness to glucose and increasing responsiveness to foliage extract. Second, we offered caterpillars a choice between disks treated with foliage extract (experimental) or solvent alone (control). The foliage-reared caterpillars approached and consumed the experimental disks disproportionately, whereas the diet-reared caterpillars approached and consumed both disks indiscriminately. This indicated that induced preferences involve olfaction and taste. Third, we ran choice tests with foliage reared caterpillars deprived of either olfactory or gustatory input. Caterpillars lacking olfactory input approached both disks indiscriminately, but fed selectively on experimental disks. In contrast, caterpillars lacking gustatory input approached experimental disks selectively, but fed indiscriminately on both types of disk. We conclude that even though olfaction helps caterpillars locate potato foliage, it is the "tuned" gustatory response that ultimately mediates the induced preference. PMID- 19308422 TI - Ultraviolet photopigment sensitivity and ocular media transmittance in gulls, with an evolutionary perspective. AB - Gulls (Laridae excluding Sternidae) appear to be the only shorebirds (Charadriiformes) that have a short wavelength sensitive type 1 (SWS1) cone pigment opsin tuned to ultraviolet (UV) instead of violet. However, the apparent UV-sensitivity has only been inferred indirectly, via the interpretation that the presence of cysteine at the key amino acid position 90 in the SWS1 opsin confers UV sensitivity. Unless the cornea and the lens efficiently transmit UV to the retina, gulls might in effect be similar to violet-sensitive birds in spectral sensitivity even if they have an ultraviolet sensitive (UVS) SWS1 visual pigment. We report that the spectral transmission of the cornea and lens of great black backed Larus marinus and herring gulls L. argentatus allow UV-sensitivity, having a lambda(T0.5) value, 344 nm, similar to the ocular media of UV sensitive birds. By molecular sequencing of the second alpha-helical transmembrane region of the SWS1 opsin gene we could also infer that 15 herring gulls and 16 yellow-legged gulls L. michahellis, all base-pair identical, are genetically UV-sensitive. PMID- 19308423 TI - Transcranial resection of a large sinonasal juvenile psammomatoid ossifying fibroma. AB - INTRODUCTION: Juvenile psammomatoid ossifying fibroma (JPOF) is a benign but potentially locally aggressive fibroosseous lesion predominantly arising in the paranasal sinuses in children and young adults. Intracranial extension is rare but occurs sometimes. In such cases, tumor resection may often require the combination of neurosurgical and facial approaches. Histological diagnosis remains a challenge because the lesion can be easily mistaken for another fibroosseous lesion or for a meningioma. CASE REPORT: We report the case of a 12 year-old boy with a JPOF arising from the right paranasal sinuses and extension towards the anterior skull base and the orbit. Despite the tumor had eroded through nasal septum, medial orbit wall, and right maxilla, it could be entirely removed performing an extended frontobasal approach via a bifrontoorbital craniotomy, obviating the need for a transfacial approach. CONCLUSION: Radiologically and histologically, the lesion could be mistaken either for a meningioma or another type of ossifying fibroma. Histological aspects and alternative surgical approaches to these rare entities are discussed. PMID- 19308424 TI - Large supra- and infra-tentorial occipital encephalocele encompassing posterior sagittal sinus and torcular Herophili. AB - Occipital encephaloceles are the most common type of encephalocele encountered in the Western Hemisphere. The occipital encephalocele is typically classified according to the relationship of the herniated tissue to the torcular. We report the unusual case of a premature infant with a rare, large occipital encephalocele which encompassed the posterior superior sagittal sinus and torcular. We discuss the variable venous anatomy with occipital encephaloceles and review the different options to employ in their surgical repair. PMID- 19308425 TI - Organized intrasylvian subarachnoid hematoma in post-traumatic child with dyskinesia for 8 years: a case report and review of the literature. AB - OBJECT: The authors present their experience with an organized intrasylvian subarachnoid hematoma (OISH) in a post-traumatic pediatric patient with dyskinesia for nearly 8 years. METHODS: An 11-year-old Chinese boy was admitted to the authors' hospital because of dyskinesia in his right upper and lower extremities. When he was 18 months old, he fell down from a trolley and then his mouth drooped to a right angle. The brain computer tomography (CT) revealed a space-occupying lesion in his left temporoparietal region. The symptom improved after 20 days of acupuncture therapy in local hospital. Two years later when he was 4 years old, his right lower limb became lame gradually with sensorial deficit. A concealed arteriovenous malformation was suggested by the brain magnetic resonance imaging and magnetic resonance angiography at that time. The child had been treated with ginkgo biloba leaf extract from 2001 to 2007 and the symptom improved gradually during that period. However, the symptom of his right upper and lower extremities deteriorated continually since January 2007. He fell down again when he was walking 1 month before he was admitted to the authors' department in July 2007. An enlarged left pterional craniotomy was performed to remove the lesion. Histopathology diagnosis was compatible with an organized hematoma with remote hemorrhage and gliosis. The child is presently healthy after 1 year's follow-up. CONCLUSION: The rarity of an OISH in a post-traumatic pediatric patient with dyskinesia for nearly 8 years makes this case very peculiar. This is the first reported pediatric case of OISH found in the literature. PMID- 19308426 TI - Epilepsy surgery could be considered a line of defense against sudden unexpected death in epilepsy. PMID- 19308427 TI - Cervical extramedullary mass lesion due to chronic CSF overshunting: case report and literature review. AB - INTRODUCTION: Several emerging clinical and neuroimaging conditions due to CSF intracranial hypotension have been documented. Recently, a few instances of a severe form of cervical myelopathy due to CSF overshunting have also been reported. Patients with this type of cervical myelopathy usually evolve with marked clinical manifestations of spinal cord involvement. MATERIALS AND METHODS: In this article, we report a 20-year-old girl, previously given a ventriculoperitoneal shunt at age 6 years, who presented with neck pain after a whiplash injury. RESULTS AND DISCUSSION: Magnetic resonance imaging revealed a cervical extramedullary mass that mimicked a cervical extradural hematoma. Further neuroimaging studies demonstrated that it corresponded to an engorged cervical epidural venous plexus that we attributed to chronic CSF overdrainage. After expectant management, the patient experienced a total recovery of her symptoms. To our knowledge, this is the first report of this condition occurring in an asymptomatic patient. Our aim in reporting this patient is twofold: (1) to warn the clinician about these neuroimaging findings that may mimic a cervical extradural hematoma and (2) to illustrate that engorged cervical epidural plexus may occur in patients in the absence of clinical manifestations of myelopathy. Recognition of this entity is important to distinguish it from a mass lesion in which a surgical intervention might be indicated. PMID- 19308428 TI - Primary intracranial leiomyosarcoma: a case report and review of the literature. AB - INTRODUCTION: Malignant tumours of the central nervous system (CNS) represent the second most common cancer type in the paediatric population of developed countries, and mortality caused by CNS tumours is the highest among paediatric cancers. Tumours of mesenchymal origin occurring either as primary or secondary lesions of the central nervous system are very rare in children. This paper describes the case of a 5-year-old non-immunocompromised male who presented with a right frontal primary intracranial leiomyosarcoma associated with large subdural collections. DISCUSSION AND CONCLUSION: Following surgical excision and adjuvant chemoradiotherapy, the patient has remained disease-free for 18 months. The literature is reviewed and the possible association of the chronic subdurals to the later development of the leiomyosarcoma explored. PMID- 19308429 TI - The right-sided aortic arch in children with esophageal atresia and tracheo esophageal fistula: a repair through the right thoracotomy. AB - PURPOSE: The management of the esophageal atresia and tracheo-esophageal fistula (EA/TEF) with right-sided aortic arch (RAA) is controversial. The preoperative diagnostic techniques may fail to show RAA associated with EA/TEF. Surgeon may need to make a decision to change the side of thoracotomy. The aim of the current study was to evaluate the possibility of preoperative diagnosis of RAA and the primary anastomosis through right chest. METHODS: A retrospective review was performed in EA/TEF patients between February 2001 and 2008. A total of 79 patients (35 female, 44 male) with EA/TEF were reviewed. Eleven (13%) patients (5 female, 6 male) had an RAA. Echocardiography was performed in 10 of 11 patients with RAA. The chest was accessed through the right side in all patients. RESULTS: The incidence of RAA was found to be higher in our study than previous studies (13%). Right thoracotomy was performed successfully in all patients. Three patients died due to multiple congenital anomalies and 1 patient due to bleeding postoperatively. Five of 10 had normal echocardiography findings. Only one patient with RAA has been successfully diagnosed by preoperative echocardiographic examination. Seven patients had no complication after operation. Their follow-up was uneventful. CONCLUSION: Preoperative recognition of RAA with echocardiography is unlikely in patients with EA/TEF but the presence of RAA does not decrease the success rate of EA/TEF repair through the right thoracotomy. PMID- 19308430 TI - Gastroschisis with jejunal and colonic atresia, and isolated colonic atresia in dichorionic, diamniotic twins. AB - Despite the increasing incidence of gastroschisis, the cause remains unknown. Genetic factors may contribute to bowel anomalies as demonstrated by cases of gastroschisis in twins and siblings, and other types of bowel anomalies in twins. Atresia of the colon represents one of the rarest causes of neonatal intestinal obstruction. We present the first case of dichorionic, diamniotic male twins in which there was gastroschisis with jejunal and colonic atresia in Twin A and isolated colonic atresia in Twin B. PMID- 19308431 TI - Supernumerary nostril together with esophageal atresia, imperforate anus and patent ductus arteriosus: a case report and review of the literature. AB - Supernumerary nostril is an extremely rare congenital entity which results from aberrant embryological development. The review of the literature reveals that only 30 cases of supernumerary nostril have been reported. We report a rare case of supernumerary nostril in association with esophageal atresia, imperforate anus, and patent ductus arteriosus. To our knowledge, this is the first case of supernumerary nostril which has been accompanied by three other congenital anomalies. In addition to the presented case, we review all the literature cases of supernumerary nostril. PMID- 19308432 TI - Survival trends in children with hepatoblastoma. AB - PURPOSE: Hepatoblastoma (HB) is a relatively rare pediatric malignancy. In this study, we present demographic data and a survival analysis from the largest patient cohort with HB reported to date. METHODS: The surveillance, epidemiology, and end results database was queried from 1973 to 2005 for all patients diagnosed with HB. Kaplan-Meier survival analysis was conducted to determine actuarial survival. Cox regression analysis was performed to determine hazard ratios (HR) for prognostic variables. RESULTS: During the study period, 459 patients with HB were identified. Overall 1-, 3-, and 5-year survival rates for the entire patient cohort were 76, 63, and 60%, respectively. Five-year survival improved over time from 36 (1973-1982) to 63% (1983-2005). Predictors of poor survival include: age 2 years and greater (HR 1.566), black race (HR 1.910), diagnosis prior to 1983 (HR 3.327), inability to perform surgical resection (HR 3.857), regional disease (HR 1.939), and distant disease (HR 3.196). CONCLUSIONS: Hepatoblastoma continues to challenge surgeons and oncologists. Most children are diagnosed early in life and undergo surgical resection whenever possible. With the advent of efficacious chemotherapy, survival has improved. Older children, black patients, and those who present with advanced disease tend to have poor outcomes. Surgical resection is the single most important predictor of survival. PMID- 19308433 TI - Quantification of rotator cuff tear geometry: the repair ratio as a guide for surgical repair in crescent and U-shaped tears. AB - INTRODUCTION: Surgical repair of symptomatic, retracted rotator cuff tears unresponsive to non-operative treatments requires closure of the tear without undue tension and reattaching the torn tendon to its former insertion site. In this study, the length of the torn tendon edge was hypothesized to be longer than the length of the humeral insertion site. The objective of this study was to quantify the discrepancy in length of the torn tendon edge and the length of the avulsed humeral insertion site. MATERIALS AND METHODS: Full thickness, rotator cuff tears that were found in twelve fresh frozen cadaver shoulders was studied. The length of the torn tendon edge, the length of the avulsed humeral insertion site and the retraction were measured using digital calipers. RESULTS: Each tear involved the supraspinatus and the infraspinatus was additionally torn in six. The size of the tear was medium in eight and large in four. The length of the torn tendon edge was always longer than the length of the avulsed humeral insertion site. Retraction was 29.9 +/- 9.3 mm (range 21-48 mm). The repair ratio, defined as the ratio of length of torn tendon edge to the length of avulsed humeral insertion site, was 2.6 +/- 0.4 (range 2.1-3.5). CONCLUSION: As only the length of the torn tendon edge equal to the length of the avulsed humeral insertion site can be repaired to bone, a repair ratio more than one precludes a simple repair and an additional repair technique such as margin convergence would be necessary for the remaining unapproximated torn tendon edge in rotator cuff tears. Repair ratio may aid in selection of the surgical repair technique of these rotator cuff tears. PMID- 19308434 TI - Densities, distribution and phenotypic expression of T cells in human fetal skin. AB - T cells are present in normal adult human skin, but their occurrence in fetal skin is unknown. T cell and Langerhans cell (LC) populations were studied using single or double immunohistochemical staining on cryostat-section. Skin samples taken from different body regions of 17 fetuses ranging from 18 to 30 weeks estimated gestational-age (w-EGA), were examined. In all specimens but one, we did not find any epidermal T cell. In contrast, dermal CD3(+) T cells occurred at all w-EGA. The density of these cells increased with increasing age. Double staining showed that CD3(+) T cells were predominantly CD4(+)/CD45RA(+). On the other hand, LC, as assessed by CD1a expression, was evenly distributed within the interfollicular epidermis and papillary dermis at all gestational ages. Analysis of T cell and LC density in different body regions did not show significant topographic differences. We suggest that lack of epidermal T cells, although the LC network was fully represented, might reflect the scarce opportunity of fetal LC to contact foreign antigens in utero. PMID- 19308435 TI - An endometriotic vault fistula presenting with monthly bleeding after hysterectomy. AB - INTRODUCTION: The aetiology of endometriosis remains unknown. The clinical presentation of endometriosis can be highly variable, occurring in numerous potential locations outside the abdomen and associated with distinct complaints. Recurrence is common, though we present a very rare case of recurrence and complication. CASE PRESENTATION: A 42-year-old caucasian woman was admitted to our unite with monthly vaginal bleeding lasting 3-5 days, beginning from 6 months after previous hysterectomy and right salpingo-oophorectomy surgery for myoma and endometrioma. We suspected of endometrioma of the left ovarium upon transvaginal ultrasonography, and diagnosed vault fistula from the endometriosis cyst to the vagina. We re-operated the patient using Pfannenstiel incision, and performed left-oophorectomy and fistula repairment. The Pouch of Douglas was obliterated and many bowel adhesions were present, indicating a stage IV endometriosis. According to our assessment, stage IV endometriosis had been present in the previous surgery. CONCLUSIONS: Considering that the short-term endometriosis recurrence is higher in premenopausal age and in advanced stage of endometriosis, bilateral oophorectomy together with hysterectomy may be a better operational choice for these patients. PMID- 19308436 TI - A prospective observational study of 363 cases operated with three different harmonic scalpels. AB - The objective of this study is to evaluate the efficacy of the Harmonic ACE, Harmonic FOCUS and harmonic scalpel with 5-mm curved blade in head and neck surgery. During a 15-month period, we performed 295 thyroidectomies, 23 parotidectomies and 45 tonsillectomies using the harmonic scalpel. Control group consisted of 106 thyroidectomies, 9 parotidectomies and 30 tonsillectomies performed with the use of conventional hemostatic techniques. The use of both Harmonic ACE and Harmonic FOCUS scalpel reduced the time of thyroid and parotid surgery by 20-25%. The use of Harmonic ACE reduced the mean time of tonsillectomy, while the use of 5-mm curved blade had no significant effect. Postoperative pain and complication rate were comparable for both the groups. In conclusion, the use of both Harmonic ACE and Harmonic FOCUS devices significantly reduces operative time in the head and neck procedures and enables a smaller neck skin incision in thyroidectomy. PMID- 19308437 TI - Imaging of electrode position in relation to electrode functioning after cochlear implantation. AB - This study assessed the electrode position in cochlear implant patients and evaluated the extent to which the electrode position is determinative in the electrophysiological functioning of the cochlear implant system. Five consecutively implanted adult patients received a multichannel cochlear implant. In all patients, the electrical impedance and the electrically evoked compound action potentials were recorded immediately after implantation. Multislice computer tomography was performed 6 weeks postoperatively before switch-on of the cochlear implant. The electrode position relative to the modiolus was assessed and correlated to the electrophysiological measurements. All electrodes were fully inserted; this was confirmed by computer tomography. The individual electrode distance toward the modiolus could be most precisely analyzed for the basal part of the electrode array. It was thus decided to study the data of electrodes one, four, and seven. No correlation was found between electrical impedance and electrode distance. A significant correlation was found between electrode distance and the electrically evoked compound action potentials, with a 96% probability using Kendall's rank correlation. We conclude that the electrode modiolus distance is of importance to the stimulation of auditory nerve fibers. Future developments in imaging will further improve and refine our insight in the relation between electrode positioning. PMID- 19308439 TI - Does balneotherapy with low radon concentration in water influence the endocrine system? A controlled non-randomized pilot study. AB - Radon bath is a well-established modality of balneotherapy for the management of degenerative musculoskeletal disorders. The present study was conducted to ascertain whether baths of relatively low (80 Bq/l) radon concentration have any influence on the functioning of the endocrine system. In the study, a non randomized pilot study, 27 patients with degenerative musculoskeletal disorders received 30-min radon baths (of 31-32 degrees C temperature and 80 Bq/l average radon concentration) daily, for 15 days. Twenty-five patients with matching pathologies were subjected to balneotherapy according to the same protocol, using thermal water with negligible radon content (6 Bq/l). Serum thyroid stimulating hormone, prolactin, cortisol, adrenocorticotropic hormone, and dehydroepiandrosterone levels were measured before and after a balneotherapy course of 15 sessions. Comparison of the accumulated data using the Wilcoxon test did not reveal any significant difference between pre- and post-treatment values or between the two patient groups. It is noted that while the beneficial effects of balneotherapy with radon-containing water on degenerative disorders is widely known, only few data have been published in the literature on its effect on endocrine functions. The present study failed to demonstrate any substantial effect of thermal water with relatively low radon content on the functioning of the endocrine system. PMID- 19308440 TI - Presumed Vogt-Koyanagi-Harada disease with unilateral ocular involvement: report of three cases. AB - AIM: To report three cases of presumed Vogt-Koyanagi-Harada (VKH) disease with unilateral ocular manifestations. METHODS: This retrospective study reviewed the long-term follow-up observations of three patients who attended the uveitis clinic at Tokyo Medical University Hospital. The patients were followed for 5-16 years with systemic clinical, ophthalmologic and laboratory examinations. Ophthalmoscopic findings, extraocular manifestations, visual acuity, and response to corticosteroid administration were evaluated. RESULTS: Three patients had characteristic clinical features of VKH involving only one eye, including diffuse choroiditis, serous retinal detachment, focal areas of delayed choroidal perfusion, multifocal areas of pinpoint leakage, macular oedema, and optic nerve staining. All patients received systemic corticosteroid therapy during the acute phase of the disease. During the follow-up period (5-16 years), all three patients developed sunset-glow fundus and nummular chorioretinal depigmented scars in the affected eye only, as well as systemic complications of deafness, vitiligo, and poliosis. CONCLUSION: The clinical and laboratory features of all three patients were typical of VKH disease except for the unilateral involvement. It is important for ophthalmologists to recognize unilateral VKH disease, even though it is a rare clinical variant of the disease. PMID- 19308441 TI - One-year results of combined photodynamic therapy and intravitreal bevacizumab injection for retinal pigment epithelial detachment secondary to age-related macular degeneration. AB - BACKGROUND: To evaluate the efficacy of combined photodynamic therapy (PDT) and intravitreal bevacizumab injection in eyes with a serous pigment epithelial detachment (PED) associated with age-related macular degeneration (AMD). METHODS: Twenty-two eyes with a serous PED exceeding two disc areas associated with AMD with choroidal vascular abnormalities [choroidal neovascularization (n = 10), polypoidal choroidal vasculopathy (n = 9), and retinal angiomatous proliferation (n = 3)] received combined PDT and intravitreal bevacizumab, and were followed about every 6 weeks for more than 1 year. Additional treatments were given for residual or recurrent lesions. The main outcome measures were changes in the PED height measured by optical coherence tomography, and the best-corrected visual acuity. RESULTS: After one treatment, the PED resolved in 12 eyes (55%) and the PED decreased in ten eyes (45%). There was no recurrence in eight (36%) eyes; however, PED recurred in 14 eyes. At 1 year, the average PED height decreased to 413 microns from the baseline 751 microns (p < 0.001). Twenty eyes (91%) had improved or stabilized vision; two eyes had decreased vision due to a retinal pigment epithelial tear and subretinal hemorrhage. CONCLUSIONS: Combined PDT and intravitreal bevacizumab may decrease the PED height and stabilize visual acuity at 1 year. PMID- 19308442 TI - Effect of the magnitude and frequency of hand-transmitted vibration on finger blood flow during and after exposure to vibration. AB - BACKGROUND: Acute exposures of the hand to vibration result in changes in finger blood flow, but it is not clear how the changes depend on the characteristics of the vibration. OBJECTIVES: To determine the effects of the magnitude and frequency of hand-transmitted vibration on finger blood flow in an exposed and a non-exposed hand during and after exposure to vibration. METHODS: Twelve persons attended seven sessions consisting of five successive periods: (1) no force and no vibration, (2) force and no vibration, (3) force and vibration, (4) force and no vibration, and (5) no force and no vibration. During the second, third and fourth periods, the palm of the right hand applied a 2-N downward force on a 25 mm diameter convex platform. During the third period, the platform was vibrated at either 16, 31.5, 63, 125, 250, or 315 Hz with the vibration magnitude increasing linearly from 0 to 15 ms(-2) rms (frequency-weighted) over 30 min (0 11 ms(-2) rms at 315 Hz). Finger blood flow was measured in the middle and little fingers of the right (exposed) hand and the middle finger of the left (unexposed) hand. RESULTS: The application of 2-N force by the palm did not affect finger blood flow on either the exposed hand or the unexposed hand. Blood flow in all three fingers (both exposed and not exposed to vibration) reduced similarly with increasing vibration magnitude, with the greatest reduction to about 40% of finger blood flow before vibration exposure. During vibration at the same frequency-weighted acceleration according to current standards, finger blood flow was dependent on the vibration frequency, with vibration at 125, 250 and 315 Hz causing the greatest reductions in finger blood flow. With vibration at 250 Hz and 315 Hz, frequency-weighted accelerations less than 1.0 ms(-2) rms reduced finger blood flow, whereas greater magnitudes were required at the lower frequencies. After the cessation of vibration, finger blood flow in exposed and unexposed fingers also depended on the vibration frequency, with greater vasoconstriction after exposure to the higher frequencies. CONCLUSIONS: During exposure to vibration, vasoconstriction in the fingers depends on both the frequency and the magnitude of the vibration. Finger blood flow was reduced by vibration magnitudes much lower than those on many powered hand tools. Vibration of one hand produced a similar vasoconstriction in fingers on the exposed and unexposed hand, suggesting a centrally mediated response. After cessation of vibration, there is continued vasoconstriction that depends on the frequency and magnitude of the prior vibration, with a similar effect in exposed and non exposed hands. PMID- 19308443 TI - Extremely small genomes in two unrelated dipteran insects with shared early developmental traits. AB - We discovered extremely small genomes (1C ~100 Mb) in the dipteran insects Coboldia fuscipes (Scatopsidae) and Psychoda cinerea (Psychodidae). The small genomes of these species cannot be explained by a fast developmental rate, which has been shown to correlate with small genome sizes in animals and plants but might accommodate the combined effects of other developmental traits, including small egg size, thin blastoderm layer, and long-germ development. PMID- 19308444 TI - Cytokeratin RNA amplification as a molecular method for the detection of lymph node metastases in breast cancer patients. PMID- 19308445 TI - The neuroscience peer review consortium. PMID- 19308446 TI - Diagnostic value of HCMV pp65 antigen detection by FCA for symptomatic and asymptomatic infection: compared to quantification of HCMV DNA and detection of IgM antibody in infants. AB - Human cytomegalovirus (HCMV) can cause symptomatic or asymptomatic infection in infants. One hundred and twenty-six infants were assessed clinically for disease in infantile period. Eighty of them were classified as symptomatic infection on the basis of physical, instrumental, and laboratory findings, 5 were demonstrated by following up to have later developed HCMV disease, and the other 41 infants were classified as asymptomatic infection. HCMV DNA was positive in all urine samples of the symptomatic infants detected by quantitative polymerase chain reaction. HCMV-IgM antibody detected by chemiluminescent immunoassay (CLIA) was positive in 62 of the 85 symptomatic infants, but was negative in all of the samples of asymptomatic infants. HCMV pp65 antigen detected by flow cytometry assay (FCA) was positive in 77 of the 85 symptomatic infants and in none of the asymptomatic infants. The coincidence to symptom of HCMV pp65 antigen detection was higher than those of HCMV DNA and HCMV-IgM antibody detection. The sensitivity, specificity, positive prognostic value and the negative prognostic value of HCMV pp65 antigen detection for diagnosis of HCMV infection was 90.6, 100, 100 and 83.7%, respectively. We concluded that detection of pp65 antigen by FCA is more sensitive for diagnosis of HCMV infection than detection of HCMV-IgM antibody and is better than HCMV DNA quantification for distinguishing the symptomatic and asymptomatic HCMV infection in infants. PMID- 19308447 TI - Pneumocystis jirovecii pneumonia in a baby with hyper-IgE syndrome. AB - A 4-month-old baby with a family history of hyper-IgE syndrome acquired Pneumocystis jirovecii pneumonia. The patient's hyper-IgE syndrome score was low, but a genetic study yielded a STAT3 mutation. P. jirovecii pneumonia can be added to the infections associated with hyper-IgE syndrome. In some cases, it may be the presenting manifestation of this immunodeficiency. PMID- 19308448 TI - Multiplanar MRI-CT fusion neuronavigation-guided serial stereotactic biopsy of human brain tumors: proof of a strong correlation between tumor imaging and histopathology by a new technical approach. AB - PURPOSE: Serial stereotactic biopsy is a diagnostic procedure, used when open biopsy or tumor bulk removal seems to be associated with a too high risk of new neurological deficits in tumors of eloquent regions or tumors of deep localizations or in anticipated high surgery related morbidity even in the older patient group. Shortcomings of this method are recognized to be the missed pathohistological information from untargeted areas in heterogeneous tumors. This study shows for the first time a collection of patients with brain tumors with their associated multiplanar MRI-CT fusion imaging during stereotaxis and the histopathological features of serial tumor biopsies along exact trajectorial sites towards the tumor center. METHODS: Thirteen patients were included. Stereotactic biopsy was performed and neuronavigation was correlated to histopathological features. RESULTS: Reactive tissue, endothelial hyperplasia, and diffusely scattered tumor cells occur outside the contrast-enhancing tumor in glioblastomas. Within the contrast-enhancing area, endothelial hyperplasia and diffuse tumor tissue were seen as compared to endothelial proliferations and the dense tumor as well as necroses in the image-defined center. CONCLUSIONS: Serial stereotactic biopsy is a reliable means. Strong correlations with the imaging characteristics of the lesions could be evaluated. PMID- 19308449 TI - Monitoring of Culicoides at nine locations in Southern Germany (2007-2008). AB - Within the countrywide monitoring of Culicoides species established by the German Federal Ministry of Food, Agriculture and User Protection (BMELV), the group Regensburg evaluated nine BG-Sentinel Midge traps located at cattle farms in the federal states Baden-Wurttemberg (BW) and Bavaria (BY) between March 2007 and May 2008. The highest numbers of midges were found in the months October and August. BTV serotype 8 genome was detected in four batches in the months September and October 2007. PMID- 19308450 TI - Feeding patterns of biting midges of the Culicoides obsoletus and Culicoides pulicaris groups on selected farms in Brandenburg, Germany. AB - Host feeding patterns of engorged sibling species of the Culicoides obsoletus and Culicoides pulicaris groups captured during three nights on two selected farms maintaining either cattle, sheep, horses, and pigs (Seedorf, Brandenburg) or cattle, sheep, moufflons, and red and fallow deer (Paulinenaue, Brandenburg) were determined by polymerase chain reaction amplification using conserved primers and sets of species-specific primers derived from vertebrates mitochondrial cytochrome b. Out of a total of 177 blood meals analysed, 115 (65%) tested positive for a blood meal from vertebrates. 63.5% (n = 73) of the cyt b positive specimens could be further assigned down to the species level. Cattle appeared to be the most attractive hosts for Palaearctic biting midges (79.5%, n = 58) even if other large vertebrates were kept in their immediate vicinity. If pigs or horses were additionally maintained on a farm, they were likewise attacked by biting midges but at a distinctly smaller rate than cattle (pigs 13.7%, horses 2.7%). In this study, game animals appear to be less attractive than cattle since only a few engorged midges had taken a blood meal from red deer (4.1%). None of the blood meals analysed tested positive for sheep. Preliminary results reveal that biting midges of the C. pulicaris and C. obsoletus groups can feed on a range of vertebrate hosts but with a distinct preference for cattle even if other livestock are maintained in adjacent areas. PMID- 19308451 TI - PCR identification of Culicoides dewulfi midges (Diptera: Ceratopogonidae), potential vectors of bluetongue in Germany. AB - Due to the severe outbreaks of bluetongue disease (BTD) in the years 2006/2007 in Germany in the absence of the main African vector Culicoides imicola, a rapid and easy applicable method for identification of autochthonous Culicoides spp. had to be developed. Morphological identification is time-consuming, rendering impossible the identification of large numbers of midges in a short period of time. A polymerase chain reaction (PCR)-based procedure in connection with a species-specific primer greatly simplifies the identification process. The region of internal transcribed spacer 1 (ITS-1) of the ribosomal DNA has shown great potential for developing a reliable PCR-based procedure. Culicoides midges were caught with ultraviolet-light traps installed on different farms in Germany during 2007 and 2008. The midges were mounted on slides and morphologically characterised. Midge DNA was extracted and the ITS-1 region amplified using conservative primers. Potential primer regions within ITS-1 were determined and a species-specific Culicoides dewulfi primer was developed to correctly identify autochthonous C. dewulfi, one of the suspected BTV vectors in northwestern Europe. The developed primer was used to identify C. dewulfi in a pool of Culicoides midges from a farm in the state of Brandenburg. PMID- 19308452 TI - Molecular cloning and analysis of stage and tissue-specific expression of Cathepsin L-like protease from Clonorchis sinensis. AB - Cathepsin L of parasite plays multiple roles in growth, food uptake, and invasion into host and pathogenesis, which makes it a valuable target for diagnosis, vaccine, and drug. In this study, we identified a cDNA encoding cathepsin L homolog (CsCPL) from the library of Clonorchis sinensis adult by bioinformatics analysis. Sequence encoding proenzyme of CsCPL (removal of signal peptide, CsproCPL) was highly expressed in form of inclusion body in Escherichia coli, and soluble rCsproCPL (about 1 mg/ml) in high purity were obtained after denaturation, purification, and renaturation. Western blot analysis indicated that CsCPL is a component of excretory-secretory products of adult, in mature form of protease. Reverse transcription polymerase chain reaction showed that CsCPL is also expressed in metacercaria and cercaria stage. Immunolocalization demonstrated that CsCPL is deposited at adult intestine, or tegument, and tegumentary cell of metacercaria and cercaria (especially at dorsal tegument of cercaria), indicating different secretory routine roles in adult and larva. The characteristics of CsCPL suggested that it may involve in invasion of cercaria into fish and development to metacercaria, excystment of metacercaria, and protein digestion of adult, which may render it a candidate antigen for fish vaccine and serodiagnosis of human clonorchiasis. PMID- 19308453 TI - Laboratory determination of efficacy of indigenous plant extracts for parasites control. AB - The present study was based on assessments of the antiparasitic activities to determine the efficacies of acetone, chloroform, ethyl acetate, hexane, and methanol dried leaf, flower, and seed extracts of Achyranthes aspera L., Anisomeles malabarica (L.) R. Br., Gloriosa superba L., Psidium guajava L., Ricinus communis L., and Solanum trilobatum L. tested against the larvae of cattle tick Rhipicephalus (Boophilus) microplus (Canestrini 1887) (Acari: Ixodidae), sheep internal parasite Paramphistomum cervi (Zeder 1790) (Digenea: Paramphistomatidae) at 2,000 ppm and fourth instar larvae of Anopheles subpictus Grassi and Culex tritaeniorhynchus Giles (Diptera: Culicidae) at 1,000 ppm. All plant extracts showed moderate effects after 24 h of exposure; however, the highest parasite mortality was found in the leaf ethyl acetate extract of A. aspera, leaf methanol extract of A. malabarica, flower methanol extract of G. superba, and leaf methanol extract of R. communis against the larvae of R. microplus (LC(50) = 265.33, 95.97, 153.73, and 181.49 ppm; LC(90) = 1,130.18, 393.88, 1,794.25, and 1,829.94 ppm); leaf acetone and chloroform of A. malabarica, flower acetone extract of G. superba, and leaf chloroform and methanol of R. communis against the adult of P. cervi (LC(50) = 108.07, 106.69, 157.61, 69.44, and 168.24 ppm; LC(90) = 521.77, 463.94, 747.02, 256.52, and 809.45 ppm); leaf ethyl acetate extract of A. aspera, leaf chloroform extract of A. malabarica, flower methanol of G. superba, and leaf methanol extract of R. communis against the larvae of A. subpictus (LC(50) = 48.83, 135.36, 106.77, and 102.71 ppm; LC(90) = 225.36, 527.24, 471.90, and 483.04 ppm); and leaf ethyl acetate extract of A. aspera, leaf chloroform extract of A. malabarica, flower methanol extract of G. superba, and leaf methanol extract of R. communis against the larvae of C. tritaeniorhynchus (LC(50) = 68.27, 95.98, 59.51, and 93.94 ppm; LC(90) = 306.88, 393.83, 278.99, and 413.27 ppm), respectively. These results suggest that the leaf ethyl acetate extract of A. aspera, leaf acetone and chloroform extract of A. malabarica, flower methanol extract of G. superba, and leaf methanol extract of R. communis have the potential to be used as an ideal eco-friendly approach for the control of the R. microplus, P. cervi, A. subpictus, and C. tritaeniorhynchus. Therefore, this study provides the first report on the larvae and adult parasitic activity of crude solvent extracts, indigenous plants consumed by the natives in southern India. PMID- 19308454 TI - MIC6 associates with aldolase in host cell invasion by Toxoplasma gondii. AB - The transmembrane microneme protein MIC6 and its partner MIC1, MIC4 comprise an adhesive complex that play important roles in host cell attachment by the obligate intracellular parasite Toxoplasma gondii. Successful penetration of host cells by T. gondii depends on coordinated interactions between MICs complex and the parasite's cytoskeleton. We have identified that the carboxy-terminal cytoplasmic domain (C domain) of MIC6 interacts with aldolase and the parasite cytoskeleton. Our finding uncovers new features regarding MIC6-aldolase interactions in host cell invasion by T. gondii. PMID- 19308455 TI - Dose-response relationships and tegumental surface alterations in Opisthorchis viverrini following treatment with mefloquine in vivo and in vitro. AB - The treatment and control of opisthorchiasis relies on a single drug, praziquantel; hence, there is a need to develop novel opisthorchicidal drugs. We investigated the in vitro and in vivo activity of the antimalarial mefloquine against Opisthorchis viverrini. Hamsters infected with O. viverrini for 2 weeks (juvenile infections) and 4 weeks (adult infections) were treated orally with single 200-400-mg/kg oral mefloquine. Worm burden reductions were assessed against untreated control hamsters. Worms were incubated in the presence of 10 and 100 microg/ml mefloquine. Scanning electron microscopy was used to examine adult O. viverrini after recovery from hamsters and following in vitro incubation. A single oral dose of 300-mg/kg mefloquine resulted in worm burden reductions of 88.5% (juvenile infection) and 96.0% (adult infections), respectively. Incubation with 10 and 100 microg/ml mefloquine resulted in rapid death of O. viverrini. Extensive tegumental disruption such as blebbing, sloughing, and furrowing was seen on worms incubated in vitro and on flukes recovered 48 h posttreatment. In conclusion, we have documented promising opisthorchicidal activities in hamsters and in vitro with the tegument being an important drug target. Proof-of-concept studies with mefloquine could be considered in opisthorchiasis patients. PMID- 19308456 TI - Synthetic nonamer peptides derived from insect defensin mediate the killing of African trypanosomes in axenic culture. AB - Synthetic antimicrobial 9-mer peptides (designated as peptides A and B) designed on the basis of insect defensins and their effects on the growth of African trypanosomes were examined using two isolates of Trypanosoma congolense, IL1180 and IL3338, and two isolates of Trypanosoma brucei brucei, ILTat1.1and GUTat 3.1, under axenic culture conditions. Both peptides inhibited the growth of all bloodstream form (BSF) trypanosomes at 200-400 microg/mL in the complete growth medium, with peptide A being more potent than peptide B. In addition, these peptides exhibited efficient killing at 5-20 microg/mL on BSF trypanosomes suspended in phosphate-buffered saline, whereas procyclic insect forms in the same medium were more refractory to the killing. Electron microscopy revealed that the peptides induced severe defects in the cell membrane integrity of the parasites. The insect defensin-based peptides up to either 200 or 400 microg/mL showed no cell killing or growth inhibition on NIH3T3 murine fibroblasts. The results suggest that the design of suitable synthetic insect defensin-based 9-mer peptides might provide potential novel trypanocidal drugs. PMID- 19308458 TI - Dynamic modeling and analyses of simultaneous saccharification and fermentation process to produce bio-ethanol from rice straw. AB - The rice straw, an agricultural waste from Asians' main provision, was collected as feedstock to convert cellulose into ethanol through the enzymatic hydrolysis and followed by the fermentation process. When the two process steps are performed sequentially, it is referred to as separate hydrolysis and fermentation (SHF). The steps can also be performed simultaneously, i.e., simultaneous saccharification and fermentation (SSF). In this research, the kinetic model parameters of the cellulose saccharification process step using the rice straw as feedstock is obtained from real experimental data of cellulase hydrolysis. Furthermore, this model can be combined with a fermentation model at high glucose and ethanol concentrations to form a SSF model. The fermentation model is based on cybernetic approach from a paper in the literature with an extension of including both the glucose and ethanol inhibition terms to approach more to the actual plants. Dynamic effects of the operating variables in the enzymatic hydrolysis and the fermentation models will be analyzed. The operation of the SSF process will be compared to the SHF process. It is shown that the SSF process is better in reducing the processing time when the product (ethanol) concentration is high. The means to improve the productivity of the overall SSF process, by properly using aeration during the batch operation will also be discussed. PMID- 19308457 TI - Mutational spectra of human cancer. AB - The purpose of this review is to summarize the evidence that can be used to reconstruct the etiology of human cancers from mutations found in tumors. Mutational spectra of the tumor suppressor gene p53 (TP53) are tumor specific. In several cases, these mutational spectra can be linked to exogenous carcinogens, most notably for sunlight-associated skin cancers, tobacco-associated lung cancers, and aristolochic acid-related urothelial tumors. In the TP53 gene, methylated CpG dinucleotides are sequences selectively targeted by endogenous and exogenous mutagenic processes. Recent high-throughput sequencing efforts analyzing a large number of genes in cancer genomes have so far, for the most part, produced mutational spectra similar to those in TP53 but have unveiled a previously unrecognized common G to C transversion mutation signature at GpA dinucleotides in breast cancers and several other cancers. Unraveling the origin of these G to C mutations will be of importance for understanding cancer etiology. PMID- 19308459 TI - Successful thrombolysis of neonatal bilateral renal vein thrombosis originating in the IVC. PMID- 19308460 TI - Fibrillary glomerulonephritis and renal failure in a child with systemic lupus erythematosus. AB - Fibrillary glomerulonephritis (FGN) is a rare immune-mediated glomerulopathy characterized by randomly arranged immunoglobulin (Ig) deposits on electron microscopy. Only seven pediatric cases have been reported, and the incidence in adults is about 1.5%. A 12-year-old boy presented with systemic lupus erythematosus (SLE) with World Health Organization Class IV lupus nephritis. A repeat biopsy carried out due to a poor response to standard immunosuppressive therapy and worsening renal functions revealed diffuse proliferative glomerulonephritis with fibrillary deposits. Despite aggressive immunosuppression with plasmapheresis and rituximab, the patient developed end stage renal disease. This is an atypical pediatric case characterized by SLE-associated FGN and a poor prognosis. PMID- 19308461 TI - Application of the new classification criteria of the Acute Kidney Injury Network: a pilot study in a pediatric population. AB - The purpose of our study was to apply the new classification criteria proposed by the Acute Kidney Injury Network (AKIN) in a pediatric population and to determine the clinical characteristics, laboratory features and outcomes of acute kidney injury (AKI) in a tertiary pediatric nephrology center in Turkey. Patients' charts from January 2003 to August 2008 were retrospectively evaluated. One hundred patients (55 male; 45 female) were enrolled. Median age at the time of AKI was 7 years (range 1 month-18 years). Patients' AKI was classified according to the staging system as follows: 25% stage 1, 36% stage 2 and 39% stage 3. The etiology of AKI was bone marrow transplantation related in 27%, renal disease in 14%, dehydration in 10%, nephrotoxic medication in 8%, cardiac surgery related in 8%, and congenital anomalies in 2%. Multiple etiologic factors with underlying chronic diseases were present in 31% of the patients. Dialysis was needed in 45% of the patients. Mortality rate was 33%. Dialysis need and mortality rate were higher in stage 2 and stage 3 patients, with a more favorable prognosis in stage 1 patients. Mortality rate was higher in patients that had undergone cardiac surgery and in those with multiple etiologic factors. The proposed AKIN staging successfully reflected the course of patients with AKI. The underlying cause of AKI seemed to be an important risk factor for death. PMID- 19308462 TI - [Therapy of patients with chronic back pain. Prescriptions--contentment- outcome]. AB - Chronic back pain is a frequent complaint in out-patient orthopaedic practice. National (DEGAM; German Society of General Medicine and Family Medicine) and international guidelines provide recommendations for diagnostic and therapeutic procedures. Regulation of the behaviour of treating orthopaedists in the context of the therapy is so far little known.In this survey it was investigated which treatments are prescribed, whether there is a link between regulations and stage of chronification and whether the subjective effects correspond to objective results. Questionnaires relating to patient anamnesis were sent to 235 orthopaedic practices and a second questionnaire was sent to all patients who took part in the survey 6 months later. A total of 630 data sets could be evaluated.It was shown that primarily passive therapies (physiotherapy, drug therapy, injections and tablets) are mostly prescribed (physiotherapy, medications, injections) and that more therapies are used with increasing stages of chronification. The prescribed therapy achieved a high subjective patient satisfaction but an objective success could not be recognized. PMID- 19308463 TI - [Amplified back pain and comorbidity in the population]. AB - BACKGROUND: Chronic back pain is rarely the sole health problem. Data concerning the association between comorbidities and different stages of chronic back pain in the German adult population are lacking. METHODS: Postal questionnaire data were collected for 3,765 persons (aged 18-75 years) in 4 German cities. Comorbidities and different stages of back pain chronification were determined. RESULTS: The prevalence and number of comorbidities varied with the extent of back pain chronification. Approximately one-third of individuals with a high degree of back pain chronicity (stages 2 and 3) received medical treatment for three or more comorbidities compared to 5% of those without amplified back pain (stage 0). CONCLUSION: Treatment concepts for chronic back pain have to consider comorbidities also present and focusing exclusively on the pain problem seems inappropriate. PMID- 19308464 TI - [Pain therapy using stimulating catheters after total knee arthroplasty]. AB - BACKGROUND: Epidural analgesia and continuous femoral nerve blocks are often used for pain therapy after total knee arthroplasty. The additional use of a continuous sciatic nerve block is controversially discussed. To avoid the problem of inaccurate placement a stimulating catheter may be useful. The aim of this study was to compare the effectiveness of a continuous femoral nerve block with an additional continuous sciatic nerve block for improved functional recovery and pain relief. MATERIAL AND METHODS: An open randomized prospective controlled study was carried out with 54 patients to receive either a stimulating catheter placed in the femoral nerve sheath or two stimulating catheters placed in the femoral and in the sciatic nerve sheath or an epidural analgesia. Pain was recorded with the visual analogue scale at rest and with passive motion of the knee during the first 3 postoperative days. The angle of nearly pain-free bending of the knee, side effects and opioid consumption were recorded. RESULTS: The reported pain scores and opioid consumption did not differ significantly between the groups. However, functional recovery up to the third postoperative day was significantly worse in the femoral catheter group. CONCLUSION: Using stimulating catheters for pain therapy the three methods are largely comparable and other parameters should be used for individual selection. PMID- 19308465 TI - Nuclear factor-kappa B decoy suppresses nerve injury and improves mechanical allodynia and thermal hyperalgesia in a rat lumbar disc herniation model. AB - Nuclear factor-kappa B (NF-kappaB) is a gene transcriptional regulator of inflammatory cytokines. We investigated the transduction efficiency of NF-kappaB decoy to dorsal root ganglion (DRG), as well as the decrease in nerve injury, mechanical allodynia, and thermal hyperalgesia in a rat lumbar disc herniation model. Forty rats were used in this study. NF-kappaB decoy-fluorescein isothiocyanate (FITC) was injected intrathecally at the L5 level in five rats, and its transduction efficiency into DRG measured. In another 30 rats, mechanical pressure was placed on the DRG at the L5 level and nucleus pulposus harvested from the rat coccygeal disc was transplanted on the DRG. Rats were classified into three groups of ten animals each: a herniation + decoy group, a herniation + oligo group, and a herniation only group. For behavioral testing, mechanical allodynia and thermal hyperalgesia were evaluated. In 15 of the herniation rats, their left L5 DRGs were resected, and the expression of activating transcription factor 3 (ATF-3) and calcitonin gene-related peptide (CGRP) was evaluated immunohistochemically compared to five controls. The total transduction efficiency of NF-kappaB decoy-FITC in DRG neurons was 10.8% in vivo. The expression of CGRP and ATF-3 was significantly lower in the herniation + decoy group than in the other herniation groups. Mechanical allodynia and thermal hyperalgesia were significantly suppressed in the herniation + decoy group. NF kappaB decoy was transduced into DRGs in vivo. NF-kappaB decoy may be useful as a target for clarifying the mechanism of sciatica caused by lumbar disc herniation. PMID- 19308466 TI - Iron-sulfur cluster biosynthesis: characterization of IscU-IscS complex formation and a structural model for sulfide delivery to the [2Fe-2S] assembly site. AB - Recent work on the bacterial iron-sulfur cluster (isc) family of gene products, and eukaryotic homologs, has advanced the molecular understanding of cellular mechanisms of iron-sulfur cluster biosynthesis. Members of the IscS family are pyridoxyl-5'-phosophate dependent proteins that deliver inorganic sulfide during assembly of the [2Fe-2S] cluster on the IscU scaffold protein. Herein it is demonstrated through calorimetry, fluorescence, and protein stability measurements that Thermotoga maritima IscS forms a 1:1 complex with IscU in a concentration-dependent manner (K(D) varying from 6 to 34 microM, over an IscS concentration range of approximately 2-50 microM). Docking simulations of representative IscU and IscS proteins reveal critical contact surfaces at the N terminal helix of IscU and a C-terminal loop comprising a chaperone binding domain. Consistent with the isothermal titration calorimetry results described here, an overall dominant contribution of charged surfaces with a change in the molar heat capacity of binding, DeltaC(p) approximately 199.8 kcal K(-1) mol(-1), is observed that accounts for approximately 10% of the total accessible surface area at the binding interface. Both apo and holo IscUs and homologs were found to bind to IscS in an enthalpically driven reaction with comparable K(D) values. Both helix and loop regions are highly conserved among phylogenetically diverse organisms from a pool of archael, bacterial, fungal, and mammalian representatives. PMID- 19308467 TI - MDMM 2008. Proceedings of the International Conference: Modeling & Design of Molecular Materials, 23-28 June 2008, Piechowice, Poland. PMID- 19308468 TI - Branchial cleft cysts in adults. Diagnostic procedures and treatment in a series of 18 cases. AB - PURPOSE: Branchial cleft anomalies may be presented as branchial cysts, fistulas, or sinuses. Purpose of this paper is to present the diagnostic procedures and the treatment in a series of branchial cleft cysts. METHODS: Eighteen patients with branchial cleft cysts were surgically treated. All of them were subjected in laboratory examinations with ultrasonography, CT or/and MRI, and fine needle aspiration cytology (FNAC). Complete excision was the treatment in all cases. RESULTS: Eight patients had Type I, seven Type II, two Type III, and one a Type IV cyst. In all cases the surgical removal was successful and after 1 to 7 years post-surgical follow-up, no recurrences have been developed. CONCLUSIONS: Branchial cleft cyst diagnostic procedure must be the same as for other neck swellings. FNAC is very useful for the diagnosis and the surgical approach must ensure safe and complete cyst removal in order to avoid intraoperative complications and recurrences. PMID- 19308469 TI - Lrrk2 R1441G-related Parkinson's disease: evidence of a common founding event in the seventh century in Northern Spain. AB - Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene together represent the most common genetic determinant of Parkinson's disease (PD) identified to date. The vast majority of patients with LRRK2-related PD reported in the literature carry one of three pathogenic substitutions: G2019S, R1441C, or R1441G. While G2019S and R1441C are geographically widespread, R1441G is most prevalent in the Basque Country and is rare outside of Northern Spain. We sought to better understand the processes that have shaped the current distribution of R1441G. We performed a haplotype analysis of 29 unrelated PD patients heterozygous for R1441G and 85 wild-type controls using 20 markers that spanned 15.1 Mb across the LRRK2 region. Nine of the patients were of Basque origin and 20 were non-Basques. We inferred haplotypes using a Bayesian approach and utilized a maximum likelihood method to estimate the age of the most recent common ancestor. Significant but incomplete allele sharing was observed over a distance of 6.0 Mb and a single, rare ten-marker haplotype 5.8 Mb in length was seen in all mutation carriers. We estimate that the most recent common ancestor lived 1,350 (95% CI, 1,020-1,740) years ago in approximately the seventh century. We hypothesize that R1441G originated in the Basque population and that dispersion of the mutation then occurred through short-range gene flow that was largely limited to nearby regions in Spain. PMID- 19308470 TI - Lehmann rotation of cholesteric droplets subjected to a temperature gradient: role of the concentration of chiral molecules. AB - We present a systematic study of the Lehmann rotation of cholesteric droplets subjected to a temperature gradient when the concentration of chiral molecules is changed. The liquid crystal chosen is an eutectic mixture of 8CB and 8OCB doped with a small amount of the chiral molecule R811. The angular velocity of the droplets strongly depend on their size and on the concentration of chiral molecules. The Lehmann coefficient is estimated by using three different methods. Our results are consistent with a Lehmann coefficient proportional to the concentration of chiral molecules. We additionally show the existence of a critical size of the droplets below which they change texture and stop rotating. PMID- 19308471 TI - Viscoelastic and failure properties of spine ligament collagen fascicles. AB - The microstructural volume fractions, orientations, and interactions among components vary widely for different ligament types. If these variations are understood, however, it is conceivable to develop a general ligament model that is based on microstructural properties. This paper presents a part of a much larger effort needed to develop such a model. Viscoelastic and failure properties of porcine posterior longitudinal ligament (PLL) collagen fascicles were determined. A series of subfailure and failure tests were performed at fast and slow strain rates on isolated collagen fascicles from porcine lumbar spine PLLs. A finite strain quasi-linear viscoelastic model was used to fit the fascicle experimental data. There was a significant strain rate effect in fascicle failure strain (P < 0.05), but not in failure force or failure stress. The corresponding average fast-rate and slow-rate failure strains were 0.098 +/- 0.062 and 0.209 +/ 0.081. The average failure force for combined fast and slow rates was 2.25 +/- 1.17 N. The viscoelastic and failure properties in this paper were used to develop a microstructural ligament failure model that will be published in a subsequent paper. PMID- 19308472 TI - A novel model-gel-tissue assay analysis for comparing tumor elastic properties to collagen content. AB - In previous work, a new assay was realized for determining soft-tissue mechanical properties. The method, named the model-gel-tissue (MGT) assay, couples material testing with a finite element model built from a micro-CT image acquisition of a gel-embedded tissue specimen to determine its mechanical properties. Given recent reports demonstrating that increased stromal collagen promotes mammary tumor initiation and proliferation, in this paper, the MGT assay is used to evaluate the modulus of murine mammary tumors and is subsequently correlated quantitatively to type I collagen content. In addition, preliminary testing of the assay sensitivity with respect to gel-volume to tissue-mass ratio is reported here. The results demonstrate a strong linear correlation between tumor mechanical properties and collagen content (R (2) = 0.9462). This result is important because mechanical stiffness as provided by the MGT assay is very similar to parameters under clinical investigation using elastographic imaging techniques. The sensitivity tests indicated that an approximate gel-volume to tissue-mass ratio threshold of 16.5 ml g(-1) is needed for successful analysis. This is an important result in that it presents guideline constraints for conducting this analysis. PMID- 19308473 TI - The global challenge of electronic waste management. PMID- 19308474 TI - A study on the carrying capacity of the available habitat for the Rhinopithecus bieti population at Mt. Laojun in Yunnan, China. AB - BACKGROUND, AIM, AND SCOPE: The Yunnan snub-nosed monkey is one of the most endangered primates in the world. It is experiencing a range of ongoing threats and the persisting effects of past disturbances. The prospects for this species are not very optimistic because habitat corridors are severely damaged by logging, grazing, and mining. Each group of the monkeys in different areas is facing a unique variety of threats. Based on genetic analysis, Rhinopithecus bieti should be separated into three management units for conservation, of which the Mt. Laojun management unit involves the most endangered primates. Despite the fact that the vegetation on Mt. Laojun is in a relatively pristine state, only two groups of monkeys, of a total of fewer than 300, survive in the area. With this paper, we aimed to address the capacity of the monkeys' habitat at the study site and the possible reasons for the small populations. MATERIALS AND METHODS: Rapid ecological assessment based on a SPOT 5 image and field survey was used to simulate the vegetation of the whole area based on reference ecological factors of the GIS system. The vegetation map of the site was thus derived from this simulation. Based on the previous studies, the three vegetation types were identified as the suitable habitat of the monkeys. The confusion matrix-based field GPS points were applied to analyze the precision of the habitat map. Based on the map of suitable habitat of the monkeys, the utilization of the habitat and the carrying capacity were analyzed in the GIS. RESULTS: The confusion matrix based field GPS points were applied to the habitat analysis process, and it was found that the habitat map was 81.3% precise. Then, with the current habitat map, we found that the mixed forest currently used by the monkeys is only a very small fraction (2.65%) of the overall potential habitat of the population, while the dark conifer forest is 4.09%. DISCUSSION: Poaching is the greatest short-term threat to this species, particularly in the southern range where local residents have a strong tradition of hunting. Quite a few individual monkeys are still trapped accidentally due to the high density of traps. These problems are hard to mitigate because it is difficult to enforce laws due to the extremely rugged terrain. CONCLUSIONS: The results show that there is a great ecological capacity of the area for the monkey's survival and a great potential for an expansion of the monkey population at the site. Based on the current population and its geographical range, it can be estimated that the suitable habitat area defined by this study can support more monkeys, about many times the current population. Thus, at least in the Mt. Laojun Area, poaching pressure is the main factor to be responsible for the low density of Yunnan snub-nosed monkeys instead of habitat alteration. RECOMMENDATIONS AND PERSPECTIVES: Based on these results, some suggestions relating to conservation can be made: Focus conservation efforts on the current distribution area of the monkeys and create a 20 km buffer zone; design a long-term plan for the suitable habitat outside the buffer zone to set up a wildlife corridor in the long run; establish an association for the local hunters exploiting, their knowledge on the animals to promote monkey conservation and stop poaching. Also, the map derived from the study helps managers to allocate conservation resources more efficiently and enhances the overall outcomes of conservation measures. PMID- 19308475 TI - Endosulfan in China 2-emissions and residues. AB - BACKGROUND, AIM, AND SCOPE: Endosulfan is one of the organochlorine pesticides (OCPs) and also a candidate to be included in a group of new persistent organic pollutants (UNEP 2007). The first national endosulfan usage inventories in China with 1/4 degrees longitude by 1/6 degrees latitude resolution has been reported in an accompanying paper. In the second part of the paper, we compiled the gridded historical emissions and soil residues of endosulfan in China from the usage inventories. Based on the residue/emission data, gridded concentrations of endosulfan in Chinese soil and air have been calculated. These inventories will provide valuable data for the further study of endosulfan. METHODS: Emission and residue of endosulfan were calculated from endosulfan usage by using a simplified gridded pesticide emission and residue model-SGPERM, which is an integrated modeling system combining mathematical model, database management system, and geographic information system. By using the emission and residue inventories, annual air and soil concentrations of endosulfan in each cell were determined. RESULTS AND DISCUSSION: Historical gridded emission and residue inventories of alpha- and beta-endosulfan in agricultural soil in China with 1/4 degrees longitude by 1/6 degrees latitude resolution have been created. Total emissions were around 10,800 t, with alpha-endosulfan at 7,400 t and beta-endosulfan at 3,400 t from 1994 to 2004. The highest residues were 140 t for alpha-endosulfan and 390 t for beta-endosulfan, and the lowest residues were 0.7 t for alpha endosulfan and 170 t for beta-endosulfan in 2004 in Chinese agricultural soil where endosulfan was applied. Based on the emission and residue inventories, concentrations of alpha- and beta-endosulfan in Chinese air and agricultural surface soil were also calculated for each grid cell. We have estimated annual averaged air concentrations and the annual minimum and maximum soil concentrations across China. The real concentrations will be different from season to season. Although our model does not consider the transport of the insecticide in the atmosphere, which could be very important in some areas during some special time, the estimated concentrations of endosulfan in Chinese air and soil derived from the endosulfan emission and residue inventories are in general consistent with the published monitoring data. CONCLUSIONS: To our knowledge, this work is the first inventory of this kind for endosulfan published on a national scale. Concentrations of the chemical in Chinese air and agricultural surface soil were calculated for each grid cell. Results show that the estimated concentrations of endosulfan in Chinese air and soil agree reasonably well with the monitoring data in general. RECOMMENDATIONS AND PERSPECTIVES: The gridded endosulfan emission/residue inventories and also the air and soil concentration inventories created in this study will be updated upon availability of new information, including usage and monitoring data. The establishment of these inventories for the OCP is important for both scientific communities and policy makers. PMID- 19308476 TI - Model-based assessment of dynamic arterial blood volume flow from ultrasound measurements. AB - To assess in clinical practice arterial blood volume flow (BVF) from ultrasound measurements, the assumption is commonly made that the velocity profile can be approximated by a quasi-static Poiseuille model. However, pulsatile flow behaviour is more accurately described by a Womersley model. No clinical studies have addressed the consequences on the estimated dynamics of the BVF when Poiseuille rather than Womersley models are used. The aim of this study is to determine the influence of assumed Poiseuille profile instead of Womersley profile on the estimation and intrasubject variability of dynamical parameters of the BVF. For this purpose, a low number of volunteers sufficed. Brachial artery centerline velocity waveform and vessel diameter were measured with ultrasound within a small group of six volunteers. Within subjects, the intra- and inter registration variability of BVF parameters estimates did not significantly differ. Poiseuille profiles compared to Womersley underestimates the maximum BVF by 19%, the maximum retrograde volume flow by 32% and the rise time by 18%. It can be concluded that when estimating in a straight vessel the dynamic properties of the BVF, Womersley profiles should preferably be chosen. PMID- 19308477 TI - Talonavicular joint arthrodesis for the treatment of pes planus valgus in older children and adolescents with cerebral palsy. AB - PURPOSE: The purpose of this report is to review our experience with talonavicular joint arthrodesis for the treatment of severe valgus foot deformities in older children and adolescents with cerebral palsy (CP). METHODS: The clinical, radiographic, and gait parameters results after talonavicular joint arthrodesis were retrospectively reviewed in 32 patients (59 feet) with valgus deformities of the foot. The surgery was performed as part of multiple simultaneous surgeries for the treatment of gait disorders. The mean age of the patients was 13.9 years (range 9-20 years) and the mean follow-up was 40 months (range 18.3-66.7 months). RESULTS: The clinical and radiographic measurements improved significantly (P = 0.000). There were no significant changes in gait parameters. Symptoms were relieved in most patients with symptomatic preoperative feet. The most frequent complication was pseudoarthrosis, which occurred in seven feet. We found a high rate of satisfaction of patients (or parents) and most of them recommended the procedure to other patients with the same condition. CONCLUSION: Talonavicular joint arthrodesis is a reliable technique that provides both functionally and cosmetically good results with a low rate of complications in the treatment of severe pes planus valgus in older children and adolescents with CP. Careful examination should rule out concomitant ankle valgus deformities. A stable fixation of the arthrodesis is recommended. PMID- 19308478 TI - Classification of proximal tibial fractures in children. AB - PURPOSE: To develop a classification system for all proximal tibial fractures in children that accounts for force of injury and fracture patterns. METHODS: At our institution, 135 pediatric proximal tibia fractures were treated from 1997 to 2005. Fractures were classified into four groups according to the direction of force of injury: valgus, varus, extension, and flexion-avulsion. Each group was subdivided into metaphyseal and physeal type by fracture location and Salter Harris classification. Also included were tibial tuberosity and tibial spine fractures. RESULTS: Of the 135 fractures, 30 (22.2%) were classified as flexion group, 60 (44.4%) extension group, 28 (20.8%) valgus group, and 17 (12.6%) varus group. The most common type was extension-epiphyseal-intra-articular-tibial spine in 52 fractures (38.5%). This study shows that proximal tibial fractures are age dependent in relation to: mechanism, location, and Salter-Harris type. In prepubescent children (ages 4-9 years), varus and valgus forces were the predominate mechanism of fracture creation. During the years nearing adolescence (around ages 10-12 years), a fracture mechanism involving extension forces predominated. With pubescence (after age 13 years), the flexion-avulsion pattern is most commonly seen. Furthermore, metaphyseal fractures predominated in the youngest population (ages 3-6 years), with tibial spine fractures occurring at age 10, Salter-Harris type I and II fractures at age 12, and Salter-Harris type III and IV physeal injuries occurring around age 14 years. CONCLUSION: We propose a new classification scheme that reflects both the direction of force and fracture pattern that appears to be age-dependent. A better understanding of injury patterns based on the age of the child, in conjunction with appropriate pre-operative imaging studies, such as computer-aided tomography, will facilitate the operative treatment of these often complex fractures. PMID- 19308479 TI - The discoid meniscus. AB - Discoid lateral meniscus is an intra-articular knee disorder typically presented in the young population and during adolescence. Different types of meniscal disorders and varied forms of presentation have been reported. The natural history depends on the type of anomaly and the presence of symptoms. Management of the disorder should be directed toward the resolution of the symptoms while preserving meniscal tissue and function. Modern surgical techniques enable suturing and preservation of meniscal tissue. The clinical manifestations, diagnostic modalities and criteria, accompanying conditions and practical management considerations are reviewed. PMID- 19308480 TI - Flexible intramedullary nailing for unicameral cysts in children's long bones : Level of evidence: lV, case series. AB - PURPOSE: The purpose of this study was to evaluate the outcome of flexible intramedullary nailing for unicameral bone cysts in terms of function and osseous consolidation. METHODS: Twenty-two unicameral bone cysts in children's long bones were treated by flexible intramedullary nailing. In 13 cases the bone cyst was diagnosed in a traumatic event leading to a pathologic fracture. Fifteen patients were referred to our clinic after failed conservative treatment. In 16 patients the cyst was located in the humerus, and in 6 patients in the femur. Mean duration of follow-up after surgery was 24 months. RESULTS: According to Capanna's criteria healing was obtained in 20 cases with a mean time of 16 months. Sixteen cysts healed completely. Four lesions were classified as grade 2, meaning that residual radiolucencies were radiographically visible at the latest follow-up. Two recurrences of humeral cysts were seen at 16 and 18 months postoperatively. The complication rate was minimal. CONCLUSION: Due to the immediate stabilization of the lesion aftercare becomes facile. This method allows prompt mobilization and early weight bearing without the necessity of a plaster cast. Further it prevents effectively the most common complication, a re fracture or a pathologic fracture. Therefore we propose this surgical procedure as the treatment of choice for unicameral bone cysts in children's long bones. PMID- 19308481 TI - Inter-observer reliability of the Stulberg classification in the assessment of Perthes disease. AB - PURPOSE: Accurate and reliable radiographic classifications are of great importance as a basis of treatment decisions and prognosis in Perthes disease. The classification of Stulberg is widely used as a predictor of long-term outcome. The aim of the present study was to determine whether the Stulberg classification is sufficiently reliable for routine clinical use in the assessment of Perthes disease. METHODS: We used this classification to assess the radiographs of 101 hips in two separate sessions (55 and 46 hips, respectively), interfered by an educational intervention in which the classification algorithm was discussed and clarified. RESULTS: We obtained good agreement between experienced examiners (weighted kappa 0.65) and a percentage agreement of 71%. We obtained weighted kappa values of 0.51 and 0.57 (moderate agreement) and percentage agreements of 62% and 65% between the least experienced observer and the two experienced examiners. Combining Stulberg class I and II, and IV and V into a simpler three-group classification gave better agreement between all observers. The agreement between the two experienced observers was improved to 81%. CONCLUSIONS: We conclude that the reliability of the Stulberg classification is acceptable when the radiographic assessment is carried out by experienced examiners. A simpler three-group classification based on the shape of the femoral head (spherical, ovoid and flat) gave better agreement and is, therefore, recommended for routine clinical use. PMID- 19308482 TI - Perthes disease: a survey of management amongst members of the British Society for Children's Orthopaedic Surgery (BSCOS). AB - The treatment of Legg-Calve-Perthes disease remains controversial. The aim of this survey was to ascertain the current management strategies of this condition amongst UK paediatric orthopaedic surgeons, with particular regard to containment procedures in the fragmentation phase. Questionnaires were distributed at the January 2006 meeting of the British Society for Children's Orthopaedic Surgery (BSCOS) and was posted to all absent members. The results showed a great deal of variability not only in the treatment of Perthes disease, but also in the decision-making processes. Consideration must now be given to a carefully constructed national multi-centre prospective randomised controlled study into the optimum management of this disease. PMID- 19308483 TI - Relapse rates following staged surgery in the treatment of recalcitrant talipes equinovarus: 9- to 16-year outcome study. AB - PURPOSE: We aimed to identify the long-term rate of relapse of deformity in a cohort of children with talipes equinovarus and to correlate it with pre operative grading. METHODS: Between 1988 and 1995, 120 club feet in 86 patients were surgically treated. A review at an average follow-up of 11.5 years (range 9 16 years) was performed. Of 59 of the children, 69 feet were assessed clinically and data for the remaining 20 children (30 feet) were obtained from the records. The patients were assessed using the method of Bensahel and Dimeglio but radiological analysis was omitted. Functional and morphological evaluation was recorded. RESULTS: Initial grades were grade 2 in 26 feet (26%), grade 3 in 48 feet (49%) and grade 4 in 25 feet (25%). Relapse occurred in one case in grade 2 (3.8%), 13 feet in grade 3 (27%) and in 19 in grade 4 (76%). Since an initial review in 1997, a further 12.5% of grade 3 and 25% of grade 4 feet have relapsed. Overall function did not correlate with severity of deformity. CONCLUSION: Relapse continues to occur after surgery during the first decade. There is a high rate of relapse (76%) in grade 4 feet. PMID- 19308484 TI - The progressive longitudinal traction reduction of unstable slipped capital femoral epiphysis: preliminary results in 11 patients. AB - PURPOSE: Unstable slipped capital femoral epiphysis is rare, but is frequently associated with avascular necrosis. The rapid and complete reduction of the epiphysis displacement was shown to be responsible for this complication. We present the preliminary results of a progressive reduction technique of epiphysis displacement. METHODS: A total of 11 patients treated using progressive traction and internal fixation were reviewed retrospectively. Only children who underwent clinical and radiological follow-up for at least 18 months were included in this study to detect avascular necrosis. RESULTS: The mean posterior displacement of the femoral head was 67 degrees and mean traction duration was 13 days. Of the patients, 2 had cutaneous problems requiring traction interruption, one on day 5 and the other on day 9. The mean residual slip was 16 degrees and 3 patients had complete or partial avascular necrosis well tolerated at the final follow-up. CONCLUSION: Our experience showed that if traction is supported for more than 2 weeks, a good correction of the epiphysis displacement can be obtained. Our short series does not allow affirmation of the superiority of our therapeutic strategy, especially with regards to avascular necrosis of the femoral head. PMID- 19308485 TI - Cross pinning of supracondylar fractures from a lateral approach. Stabilization achieved with safety. AB - PURPOSE: Various pin configurations are possible to stabilize a supracondylar fracture. While cross pinning gives the best stabilty the disadvantage is the risk of iatrogenic ulnar nerve injury. We combine a cross pin fixation with a lateral approach. The aim of the study was to prove our method retrospectively to show the advantage of lateral cross-pinning achieving stabilty and avoiding ulnar nerve injury. METHOD: Between 1 January 2001 and 31 December 2002, 84 supracondylar fractures were treated with invasive surgical procedure. The intention was a primary closed reduction. Following closed reduction under general anaesthesia, two K-wires were introduced from the lateral side, one ascending and the other descending. If it was not possible to perform a closed reduction, an open reduction was performed by medial-lateral crossed K-wire fixation. After either an open or closed reduction, the pins were buried under the skin. The results were evaluated using Flynn's score. The mean time of follow up was 18.9 months. RESULTS: Seventy-seven percent of the patients were treated with a closed reduction, while 23% needed an open reduction. A clinical follow-up examination was done at an average of 18.9 months following the trauma. Of those treated with a closed reduction alone, 93% had an excellent or good functional result. Of those requiring an open reduction, 88% had excellent or good result. None of our patients exhibited secondary dislocation or iatrogenic ulnar palsies. CONCLUSION: Closed reduction and lateral crossed pin fixation with ascending and descending K-wires buried under the skin is an effective method to treat type II and III supracondylar fractures in children. The method gives stability and avoids iatrogenic ulnar nerve injuries. PMID- 19308486 TI - Results after surgical treatment of thumb duplication: a retrospective review of 33 thumbs. AB - PURPOSE: Purpose This study reports the results of surgical treatment of thumb duplication in the Clinique d'Orthopedie et de Chirurgie de l'Enfant de l'Hopital Jeanne de Flandre in Lille (France). Methods Thirty patients (33 thumbs) operated on between 1995 and 2003 are clinically reviewed. Results The mean postoperative follow-up was 3 years and 11 months. According to Wassel's classification, the series included 12 type II duplications, two type III, 14 type IV, two type V, one type VI and two type VII. The surgical approaches consisted of simple resection of the most hypoplastic thumb (16 thumbs), the Bilhaut-Cloquet procedure (ten thumbs) and resection associated with reconstructive surgery (seven thumbs). The Bilhaut-Cloquet procedure was used in three cases for treatment of type IV duplication On the basis of the Tada scoring system, we obtained 24 good results, eight fair results and one poor result. Conclusion Based on our results, we recommend that the Bilhaut-Cloquet procedure be used not only for the treatment of type II duplication when the thumbs are both hypoplastic and symmetric but also for type IV duplication with the same clinical parameters. For the other types of duplications, we consider that resection of the most hypoplastic thumb associated with reconstructive surgery is the best surgical approach. For type VII duplication, ablation of the triphalangeal thumb remains the best option. We do not recommend osteotomy at the first surgery. PMID- 19308487 TI - The spectrum of hand and foot malformations in patients with Greig cephalopolysyndactyly. AB - PURPOSE: Greig cephalopolysyndactyly (GCPS) (OMIM 175700), a rare autosomal dominant disorder, is characterized by a distinct combination of craniofacial, hand and foot malformations. The hand and foot malformations often require orthopedic assessment and treatment. The disorder is caused by point mutations or deletions in the GLI3 gene, located on chromosome 7p14.3. Herewith, we review the hand and foot malformations in a cohort of 13 patients referred for genetic testing. METHODS: We reviewed the medical files of 13 patients with GCPS seen at the Center for Human Genetics in Leuven between 2003 and 2005. Clinical, molecular and radiological findings, when available, were recorded. RESULTS: We identified six different point mutations in the GLI3 gene, two microdeletions and three larger chromosomal deletions. In the hands, preaxial polydactyly was never observed, but the malformations included postaxial polydactyly, broad thumbs, clinodactyly of the thumbs and various degrees of syndactyly. In the feet the spectrum of malformations included preaxial polydactyly, postaxial polydactyly, different degrees of syndactyly and broad halluces. Syndactyly of the toes and hallux abnormalities were present in all patients. Most frequently, syndactyly was present between toes 1-2-3. The broadening of the hallux was either due to a complete or partial duplication of the first toe or to broadening of the distal phalanx. Mental retardation was found in three cases and was associated with a large chromosomal deletion of the GLI3 region. CONCLUSION: We found the classic hand and foot malformations associated with GCPS in our cohort of patients. Patients with a large chromosomal deletion had mental retardation, but no structural brain anomalies were found. PMID- 19308488 TI - A technique for calculating limb length inequality and epiphyseodesis timing using the multiplier method and a spreadsheet. AB - PURPOSE: Limb length inequality and epiphyseodesis timing calculations are common in pediatric orthopedics. The multiplier method developed by Paley et al. has significantly simplified the calculation of ultimate limb length. The calculation of limb length inequality has also become very simple for congenital limb length inequalities. However, the equations for limb length inequality from acquired differences and epiphyseodesis timing are not simple, and are prone to arithmetic errors. METHODS: To limit these errors, we have developed a spreadsheet which finds the appropriate multipliers, solves the equations, and computes the predicted limb length inequality and the appropriate age for an epiphyseodesis. RESULTS: This method of using a spreadsheet and the multiplier calculations is quite simple and quick to use in practice. The calculations can be cut and pasted into computerized charts for future reference. CONCLUSIONS: We recommend this method for clinical use and make it available for public use. PMID- 19308489 TI - Predicting the outcome of Legg-Calve-Perthes' disease in children under 6 years old. PMID- 19308490 TI - A new approach to the treatment of congenital vertical talus. AB - Congenital vertical talus is an uncommon foot deformity that is present at birth and results in a rigid flatfoot deformity. Left untreated the deformity can result in pain and disability. Though the exact etiology of vertical talus is unknown, an increasing number of cases have been shown to have a genetic cause. Approximately 50% of all cases of vertical talus are associated with other neuromuscular abnormalities or known genetic syndromes. The remaining 50% of cases were once thought to be idiopathic in nature. However, there is increasing evidence that many of these cases are related to single gene defects. Most patients with vertical talus have been treated with major reconstructive surgeries that are fraught with complications such as wound necrosis, talar necrosis, undercorrection of the deformity, stiffness of the ankle and subtalar joint, and the eventual need for multiple operative procedures. Recently, a new approach to vertical talus that consists of serial casting and minimal surgery has resulted in excellent correction in the short-term. Longer follow-up will be necessary to ensure maintenance of correction with this new technique. A less invasive approach to the correction of vertical talus may provide more favorable long-term outcomes than more extensive surgery as has been shown to be true for clubfoot outcomes. PMID- 19308491 TI - History of the functional method for conservative treatment of clubfoot. PMID- 19308492 TI - Beware of ulnar nerve entrapment in flexion-type supracondylar humerus fractures. AB - PURPOSE: A recent study reported a higher incidence of pre-operative ulnar nerve symptoms in patients with flexion-type supracondylar fractures than in those with the more common extension supracondylar fractures and a greater need for open reduction (Kocher in POSNA paper #49 2006). We have encountered a specific pattern of flexion supracondylar fractures that often require open reduction with internal fixation (ORIF) due to entrapment of the ulnar nerve within the fracture. METHODS: Medical records and X-rays from 1997 to 2005 at our children's hospital were examined to identify flexion supracondylar fractures that required open reduction. The operative reports were reviewed to identify cases that had the ulnar nerve blocking the reduction. RESULTS: During the 8 years examined, 1,650 supracondylar fractures had been treated by means of closed reduction and percutaneous pinning. Of these, only 1.8% or 30 cases could not be reduced closed and required open reduction internal fixation, excluding 11 open fractures. Of the 30 fractures requiring open reduction internal fixation, 24 were of the extension type needing ORIF because of interposed periosteum/muscle. The other 6 patients had flexion-type supracondylar fractures that failed closed reduction. All had a persistent medial gap at the fracture site. All 6 fractures had interposed periosteum or muscle, while in 3 cases the ulnar nerve was also entrapped within the fracture site (Figs. 1, 2) CONCLUSION: Flexion-type supracondylar fractures remain a relatively uncommon variant (2-3%) of supracondylar fractures. Recent reports have noted that open treatment of these fractures is required more frequently than for extension fractures. In our series, 20% of the open cases were flexion-type fractures and in half of these the ulnar nerve was found to be entrapped in the fracture, preventing reduction. PMID- 19308493 TI - Screw placement in slipped upper femoral epiphysis: is good the enemy of better? AB - Introduction and aims A single hip screw is the recommended method of fixation for slipped upper femoral epiphysis (SUFE). Current practice favours the placement of the screw in the centre of the femoral head on both anteroposterior and lateral planes to avoid the risks of chondrolysis and avascular necrosis (AVN). We investigated the correlation between different positions of the screw in the femoral head and the prevalence of AVN, chondrolysis, late slippage and the time to epiphyseal closure. Methods The clinical notes and radiographs of 38 consecutive patients (61 hips) who underwent single screw fixation for SUFE were evaluated retrospectively with a mean follow-up of 36 months. Two-way ANOVA and the post hoc test was performed to analyse the correlation between the different variables and the outcome at the 5% level of significance. Results There were 16 acute slips, 18 chronic slips and ten acute-on-chronic slips. Seventeen slips were treated prophylactically. Mild slip was encountered in 39 hips, moderate slip in four and severe slip in one. The central-central position was only achieved in 51% of cases. The most significant results of the study were as follows: (1) no significant difference between the time to epiphyseal closure and the position of the screw, and (2) no late slippage or chondrolysis was observed in our series. Conclusion Our results showed that the positioning of the screw other than in the centre of the femoral head has the ability to provide physeal stability and has no correlation with the timing to closure of the epiphysis and the risk of avascular necrosis or chondrolysis. We therefore recommend that other positions be considered if the "optimal central-central position" is not initially achieved - specifically for the treatment of mild slip - as the potential hazards from several attempts to achieve the optimum position outweigh the benefits. PMID- 19308494 TI - Development and evaluation of thoracic kyphosis and lumbar lordosis during growth. AB - PURPOSE: The aim of this study was to establish ranges of angular variation in lordotic and kyphotic curves in normal male and female children and adolescents. METHODS: We developed a pantograph to measure dorsal curves. It consisted of a tripod-supported vertical strut to which an articulated bar was fixed and which had an arm that was able to follow the dorsal surface while moving up and down. This arm was positioned over the C7 spinous process and followed spinous processes to L5 at constant speed. A laser beam was used to ensure the proper positioning of the pantograph and the subject. The motion was recorded using software so that the dorsal outline was represented on a computer screen, and lordotic and kyphotic curves were automatically measured. Before performing the population study, the pantograph was validated in 20 normal subjects by comparing the pantograph measurements with lateral spine radiographs. There were no statistically significant differences in the measurements. There were 718 subjects with no race selection, of whom 350 were males and 368 females ranging in age from 5 to 20 years and presenting normal weight and height. Individuals with generalized ligament laxity, trunk asymmetry, muscle retraction, or any orthopedic anomaly were excluded from the study. Data were analyzed according to age and gender. Student's t tests and regression analysis were performed. RESULTS: Kyphotic curves increased linearly from 25 degrees at 7 years of age to 38 degrees at 19 years of age (kyphotic angle = 25 degrees + 0.58 x age). Lordotic curves increased linearly from 22 degrees at 5 years of age to 32 degrees at 20 years of age (lordotic angle = 24 degrees + 0.51 x age). There were no differences between males and females. CONCLUSIONS: The pantograph that was developed for this study was successfully used to establish the normal ranges and progression of thoracic kyphosis and lumbar lordosis in the studied population. Both curves increased linearly with age, with no differences between males and females. PMID- 19308495 TI - Soft-tissue sarcomas of the extremities in patients of pediatric age. AB - PURPOSE: The extremity site is a peculiar location for soft-tissue sarcomas (STS) of children and adolescents. Here we report the experience of the Pediatric Oncology Unit of the Istituto Nazionale Tumori of Milan, Italy, concerning 204 patients with STS of the limbs treated between 1977 and 2006. METHODS: The study series included 52 patients with rhabdomyosarcoma (RMS)(65% of which were of the alveolar subtype), nine with extraosseous Ewing sarcoma and 143 with non rhabdomyosarcoma soft-tissue sarcomas (NRSTS), 38% of which were synovial sarcoma. Patients were treated with a multimodality approach including surgery, chemotherapy, and radiotherapy. RESULTS: For the RMS patients, the 5-year event free survival (EFS) rate was 37.1%, with distant metastases being the main cause of treatment failure. The outcome was particularly poor for patients with large invasive tumors, hand/foot involvement and/or nodal or distant metastases, and for patients who were not given radiotherapy. For the extraosseous Ewing sarcoma cases, 5-year EFS was 74%. For the NRSTS cases, the 5-year EFS was 72.6%: tumor size and local invasiveness, tumor grade, malignant peripheral nerve sheath tumor (MPNST) histology, and distant metastases were the main prognostic factors. DISCUSSION: While the limbs are the most common sites of NRSTS and are often characterized by a more favorable prognosis than for axial tumors, the clinical features of extremity RMS often differ from those of RMS of other sites, with a higher incidence of unfavorable prognostic factors (e.g., alveolar subtype) and consequently unsatisfactory treatment results. The treatment of these patients is complex and necessarily multidisciplinary, and it demands not only adequate experience of treating children and adolescents in clinical trials, but also particular skills in the field of orthopedic surgery. PMID- 19308496 TI - Associated risk factors in children who had late presentation of developmental dysplasia of the hip. AB - PURPOSE: The purpose of this study was to assess the role of clinical examination, associated risk factors and plain radiograph of the pelvis in children who had late presentation of DDH. METHODS: We report on a 7-year prospective study, in children who had late presentation of developmental dysplasia of the hip (DDH). For this purpose, 740 hips in 370 referred children, age range 3-7 months (mean 3.44 months) were clinically and radiologically assessed, and the associated risk factors recorded. RESULTS: Female sex, first born, positive family history and breech presentation were confirmed as risk factors for DDH. Significant findings were an increased risk for vaginal delivery over caesarean section for breech presentation (P = 0.002). There was an increased risk for caesarean section in the absence of breech presentation. Multiple births and preterm births had a reduced risk. For breech presentation, the risk of DDH was estimated to be at least 1.6% for girls and 3.4% for boys; a combination of factors increased the risk. Limitation of abduction (43.2%) and asymmetry of the groin skin folds (72.7%) were found to be the two most common clinical findings associated with DDH. Bilateral acetabular dysplasia is more common than unilateral dysplasia. Foot deformities were rarely encountered in children with acetabular dysplasia. CONCLUSIONS: The percentage of first-born babies who had DDH is lower than reported in the literature (34%), but still shows significant risk. We did find that bilateral acetabular dysplasia is more common than unilateral dysplasia. Torticollis and foot deformities are rarely found to be associated with DDH. All these findings needs further evaluation in children who had surgical treatment for DDH, to see if they are different from dysplastic groups. Limitation of abduction is an important clinical finding, but is not always associated with DDH. Asymmetry of the skin folds in the groin were found to be an important clinical finding associated with DDH for all age groups. As clinical examination depends on many factors, and most DDH cases are of the dysplastic type, it is mandatory to depend on further diagnostic tools for confirmation of DDH. PMID- 19308497 TI - Differences in biomechanical stability of femur fracture fixation when using titanium nails of increasing diameter. AB - PURPOSE: The purpose of this study was to compare the biomechanical stability generated when utilizing increasing sizes of titanium (Ti) flexible nails for fixation of simulated comminuted femur fractures. METHODS: Five synthetic adolescent-sized femur models were reamed to create a 9-mm canal. A 2-cm section was removed in the mid-diaphysis to simulate comminution. Each femur was first stabilized with bilateral, retrograde 3.0-mm titanium elastic nails. Femurs were tested in axial rotation and axial compression. The constructs were removed, and femurs were re-nailed with 3.5-mm nails. Identical testing was conducted. These nails were then removed, and femurs were re-nailed with 4.0-mm nails. This provided data on "canal fill" representing 67, 78 and 89% of the reamed canal diameter. Data for axial rotation (degrees) and failure load (N) required to produce 5 mm of fracture shortening were analyzed with a one-way ANOVA (P < 0.05) and a Tukey's post-hoc test for multiple comparisons. RESULTS: For axial rotation, there were statistically significant improvements in rotational control for each increase in nail size. For axial stability, each increase in nail size resulted in increased axial failure loads to 5 mm, although these data were not statistically different. A specific comparison between 3.0- and 3.5-mm nails for compressive stability found significantly greater stability afforded by using 3.5 mm nails. CONCLUSIONS: Data from this study demonstrate that increasing the amount of canal fill provides significant improvements in rotational control. The largest improvement was seen when increasing from 3.0- to 3.5-mm nails. While increasing the nail size from 3.5 to 4.0 mm again provided greater stability, larger nails may be more difficult to insert. Thus, increasing the nail size for femoral fracture fixation should be considered after measuring the diameter of the canal and evaluating the potential difficulty of insertion as well as specific demands of the fracture pattern. PMID- 19308498 TI - A technique for calculating limb length inequality and epiphyseodesis timing using the multiplier method and a spreadsheet. PMID- 19308499 TI - Coming full circle the high-tech way. PMID- 19308500 TI - Current approach to fibrous dysplasia of bone and McCune-Albright syndrome. AB - Fibrous dysplasia (FD) of bone is an uncommon disease caused by sporadic, congenital mutations in the cAMP regulating protein, G(s)alpha. It is an example of somatic mosaicism in which a wide spectrum of disease is possible. Widespread skeletal involvement is often associated with varying combinations of cafe-au lait skin spots, and/or endocrine dysfunction (precocious puberty, renal phosphate wasting, hyperthyroidism, and/or growth hormone excess). Unrecognized and untreated endocrine dysfunction can exacerbate the skeletal disease. The diagnosis is usually established on clinical grounds on the basis of physical examination and typical radiographic appearance. Occasionally, gene testing of affected tissue may be helpful. The skeletal sites involved with disease are established at an early age, and the complications of fracture deformity are most pronounced in childhood. Bone pain in the absence of a fracture is more common in adults, but can also be present in children. Treatment with bisphosphonates is usually effective at relieving pain, but probably has no effect on the natural history of the disease. Scoliosis, which was previously thought to be an uncommon occurrence, has been shown to be common and progressive, and as such, warrants investigation and, when necessary, surgical treatment. The surgical management of FD remains challenging. Timing and technique remain controversial, but some consensus exists in that grafting materials (of any type) usually fail and should not be a central aspect of the surgical approach. Intramedullary devices are in general superior to side plates and screws. In extremely widespread disease with very early fracture and deformity, no surgical approach will affect final functional outcome. Efforts should be made for the initiation of international collaborative studies to better define optimal surgical approaches to the treatment of this challenging disease. PMID- 19308501 TI - The management of Legg-Calve-Perthes' disease: is there a consensus? : A study of clinical practice preferred by the members of the European Paediatric Orthopaedic Society. AB - PURPOSE: The aim of the study was to find out whether or not there is consensus among experienced pediatric orthopaedists about the management of certain clinical scenarios in Legg-Calve-Perthes' disease. METHODS: A questionnaire was sent to all 297 members of the European Paediatric Orthopaedic Society (EPOS) describing four cases of Legg-Calve-Perthes' disease (LCPD) with two X-rays each and a short description of the clinical scenario. Two of the patients were younger and two were older than six years of age. From both age groups there was one with a good range of motion and an X-ray classified as Herring A or B, while the other patient had a poor range of motion and an X-ray classified as Herring C. EPO members were asked to choose from various treatment options or to describe any other therapy that they would advise in the clinical scenarios. RESULTS: One hundred and fifty members answered the questionnaire. The participants had an average of 20 years of experience in pediatric orthopaedics. There was a consensus that no surgery should be performed in a young patient with a good range of motion and that there should be no weight relief when older with a good range of motion. Conservative containment treatment (abduction splint, Petrie cast) and arthrodiastasis was suggested in only very few centres. There was a tendency to perform an operation when the patient is older with a poor range of motion and to perform operative treatment only when there were subluxation or head at risk signs. pelvic osteotomies or a combination of pelvic and femoral osteotomies rather than femoral osteotomies alone. Age did not determine the indication for treatment and there was no agreement on the indications for physiotherapy. There was also no consensus on the type of pelvic osteotomy to be used. CONCLUSIONS: The study showed that indications for the treatment of LCPD is based more on the personal experience of the surgeon rather than on scientific data. PMID- 19308502 TI - Predicting the outcome of Legg-Calve-Perthes' disease in children under 6 years old. AB - PURPOSE: In this study, we investigated the outcome of Legg-Calve-Perthes disease in children presenting under 6 years old. Firstly, we wished to know the outcome in terms of femoral head deformity in this age group. Secondly, we investigated the potential for improvement in femoral head deformity with growth and remodelling after the Perthes lesion had healed. Thirdly, we assessed the ability of Herring's lateral pillar classification to predict femoral head deformity in this age group. METHODS: The outcome in this group was assessed using Stulberg's classification of femoral head deformity and compared to that for older children with Perthes seen at our institution over the same period. To investigate the potential for improvement of Stulberg grade with growth we assessed the hips at the stage of final healing (Waldenstrom IV) and again at skeletal maturity. To assess whether Herring's classification of lateral column collapse is predictive of deformity at this age we recorded the Herring class A, B or C at the time of maximum fragmentation and compared this to Stulberg grade. RESULTS: Out of a total of 127 children (131 hips) seen at our institution, 67 children (69 hips) presented before the age of 6 years. The number of Stulberg III and IV hips was 14 (20%) and 10 (15%), respectively. Chi-squared test of age against outcome confirmed that the younger children overall had a better outcome (P < 0.001). Stulberg grade did not improve predictably after the stage of final healing. Twenty-three hips graded as Stulberg III and IV at Waldenstrom IV were reassessed at skeletal maturity. Three changed from Stulberg III to IV and five changed from Stulberg IV to III, but none had improved to Stulberg I/II. Herring's classification was predictive of outcome, with 96% of Herring A hips having Stulberg I or II outcome, 61% of Herring B hips having Stulberg I or II outcome, while 60% of Herring C hips (18 hips) had significant femoral head deformity, and 10 hips Stulberg III and 6 hips Stulberg IV. CONCLUSIONS: Perthes in children presenting before 6 years of age can result in significant femoral head deformity which does not predictably improve with time. The risk of a poor outcome can be predicted by Herring's classification of lateral pillar collapse. PMID- 19308503 TI - Hip range of motion in Perthes' disease: comparison of pre-operative and intra operative values. AB - BACKGROUND: Children with early Perthes' disease and clinical loss of motion are commonly noted to have normal or almost normal hip range of motion (ROM) when examined under anesthesia (EUA). Despite anecdotal comments about this observation, no series has documented these findings. The lack of such data led to this study. METHODS: Consecutive patients with unilateral Herring class B or C Perthes' disease in mid to late Waldenstrom's fragmentation stages who failed a non-operative course of hip motion restoration had hip range of motion (ROM) documented pre-operatively and intra-operatively by a single, experienced pediatric orthopedic surgeon. The opposite normal hip's ROM was used as the control. Joint congruity was assessed arthrographically. RESULTS: Twenty-seven patients, 6-10 years old (average 7.9 years) with early Perthes' disease were reviewed. Twenty-one of 27 patients (77.7%) had full or almost full hip ROM of the involved side when examined under anesthesia (EUA). Six patients (22.3%) had limited abduction (<50 degrees) and underwent an adductor tenotomy that normalized hip ROM. Five of these six patients were older than 8 years. Intra operative arthrography did not demonstrate lack of containment, hinge abduction or joint incongruity. CONCLUSIONS: Loss of hip ROM in this series of patients with early Perthes' disease without intra-articular incongruity was due to pain and muscle spasm which were obviated by anesthesia, demonstrating essentially normal hip ROM in the majority of patients. Pre-operative bed rest and traction was not required to restore ROM in this group of patients. PMID- 19308504 TI - Comparison of rectus femoris transposition with traditional transfer for treatment of stiff knee gait in patients with cerebral palsy. AB - PURPOSE: To assess the outcome of children with cerebral palsy following reposition of the distal rectus femoris tendon for treatment of stiff knee gait. METHODS: Children with cerebral palsy with stiff knee gait who underwent rectus femoris transfer were studied retrospectively. Inclusion criteria were cerebral palsy of diplegic or quadriplegic type, preoperative and 1 year postoperative three-dimensional motion analysis, and no other surgery except rectus femoris transfer at the time of study. The patients were separated into two groups: in group I, the rectus femoris was transferred to the distal medial hamstring tendons, either the gracilis or the semitendinosus; in group II, the distal tendon of the rectus femoris was transposed laterally and attached to the iliotibial band/intermuscular septum. RESULTS: Peak knee flexion during swing phase, total dynamic knee range of motion, knee range of motion during swing phase, and time to peak knee flexion during swing phase were all improved in both groups. Hip and pelvic kinematics were not influenced by the surgery. Velocity, stride length, and cadence were all improved following the surgery. There was no difference between the transfer group and the transposition group. CONCLUSION: These findings suggest that distal transfer of the rectus femoris is effective in improving swing phase knee function by diminishing the mechanical effect of the dysphasic swing phase activity of the rectus femoris, not by converting the rectus femoris to an active knee flexor. PMID- 19308505 TI - Radiographic threshold values for hip screening in cerebral palsy. AB - PURPOSE: To analyse the consequences of using different radiographic measurements and different threshold values for hip screening in children with cerebral palsy (CP). METHODS: In a total sample of children with CP a standardised radiological follow-up of the hips was carried out as a part of a hip prevention programme. Acetabular index (AI) and migration percentage (MP) were measured on all radiographs. In this study, 1,067 radiographs of 272 children born 1992-1998 were analysed. RESULTS: Lateral displacement of the femoral head was common without acetabular dysplasia, and acetabular dysplasia occurred at a later stage than femoral head lateralisation. Hip dysplasia without lateral displacement of the femoral head was rare. In 16 of the 56 hips (29%) with AI >/= 27 degrees and in 23 of the 71 (32%) hips with MP >/= 33% the values decreased below the threshold value without operative treatment. In hips with AI >/= 30 degrees only 2 of 31 hips (6%) and in hips with MP >/= 40% only 5 of 44 hips (11%) decreased below the threshold values without operative treatment. CONCLUSIONS: Radiographic follow-up with only measurement of the MP seems sufficient in screening for dislocation in children with CP. MP >/= 33% is recommended as threshold for reaction or intensified observation. In children with MP >/= 40%, the lateral displacement increased over time in most hips, thus indicating the need for operative intervention. In children with MP 33-40%, treatment should be based on other clinical signs and the progression of MP over time. PMID- 19308506 TI - Pre-axial mirror polydactyly associated with tibial deficiency: a study of the patterns of skeletal anomalies of the foot and leg. AB - PURPOSE: The study was undertaken to identify the patterns and spectrum of aberrant development of the skeletal elements of the leg and foot in children with pre-axial mirror polydactyly of the foot. METHODS: Case records and radiographs of eight children (ten feet) with pre-axial mirror polydactyly were studied. Four feet were seen in association with fibular dimelia, three feet with dysplastic trapezoid shaped tibia, two with tibial aplasia and one with tibial hypoplasia. RESULTS: Distinct patterns of anomalies of the hindfoot, midfoot and forefoot were noted, with different patterns of leg malformation. Fibular dimelia was associated with duplication of the calcaneum, cuboid and the lateral cuneiform. Talar duplication occurred in children with a trapezoidal tibia. No duplication of tarsal bones was seen in association with tibial aplasia or hypoplasia. The first metatarsal was hypoplastic wherever the tibia failed to develop. Mere resection of the supernumerary rays resulted in a near normal looking foot in feet without calcaneal duplication. The cosmetic appearance of the foot was poorest in children with fibular dimelia. PMID- 19308507 TI - Accuracy of complex lower-limb deformity correction with external fixation: a comparison of the Taylor Spatial Frame with the Ilizarov ring fixator. AB - PURPOSE: Circular external fixators have several advantages over other surgical options in the treatment of limb length discrepancy and axial deformity. The innovative Taylor Spatial Frame (TSF) combines a rigid hexapod fixation system with the support of a web-based software program, and thus offers the possibility of simultaneous corrections of multidirectional deformities. Whilst there is still some scepticism of many Ilizarov device users about the advantages of the TSF, the purpose of the study was to perform a comparison between the TSF and the Ilizarov ring fixator (IRF) with regard to the accuracy of deformity correction in the lower limb. METHODS: Two hundred and eight consecutive deformity corrections in 155 patients were retrospectively evaluated. There were 79 cases treated with the IRF and 129 cases treated with the TSF. The mean age of the patients at the time of surgery was 13.2 years (range; 2-49 years). Standing anteroposterior and lateral radiographs were evaluated preoperatively and immediately after removal of the frames. The final result was compared to the preoperatively defined aim of the deformity correction. According to the treated count of dimensions, we differentiated four types of deformity corrections. The results were graded into four groups based on the persisting axial deviation after removal of the frame. RESULTS: The aim of the deformity correction was achieved in a total of 90.7% in the TSF group, compared to 55.7% in the IRF group. On the basis of the count of dimensions, the TSF achieved obviously higher percentages of excellent results (one dimension: TSF 100%; IRF 79.3%; two dimensions: TSF 91.8%; IRF 48.6%; three dimensions: TSF 91.1%; IRF 28.6%; four dimensions: TSF 66.7%; IRF 0%). In addition, the degree of the persisting deformity increased with the number of planes of the deformity correction. CONCLUSIONS: The TSF allowed for much higher precision in deformity correction compared to the IRF. In two-, three- and four-dimensional deformity corrections in particular, the TSF showed clear advantages. This may derive from the TSF specific combination of a hexapod fixator with the support of an Internet-based software program, enabling precise simultaneous multiplanar deformity corrections. PMID- 19308508 TI - Acute lymphoblastic leukemia in children: correlation of musculoskeletal manifestations and immunophenotypes. AB - PURPOSE: Studies on musculoskeletal manifestations (MSM) of childhood acute lymphoblastic leukemia (ALL) have yielded variable findings with regard to their clinical impact. We investigated the significance for differential diagnosis, treatment and outcome of musculoskeletal complaints as presenting symptoms of ALL, and their correlation with leukemia immunophenotypes, for which data is lacking. METHODS: Data on 783 children in the national study for childhood ALL between 1984 and 2003 were reviewed retrospectively. Statistical analysis examined possible relationships between MSM at the time of diagnosis and demographic and clinical data, biological features of leukemia (peripheral blood counts, immunophenotype and main cytogenetic aberration), response to initial prednisone treatment, and outcome. RESULTS: Of 765 children with data on orthopaedic complaints, 240 presented with MSM (31.4%). Among these children, B cell precursor (BCP) was much more common (209/576, 36.3%) than T cell ALL (25/176, 14.2%). Patients with MSM had lower white blood cell counts (WBC) (median of 9 vs. 20 x 10(9)/L, P < 0.001) and percentage of blast cells in the peripheral blood at diagnosis compared to those without (median of 27 vs. 53%, P < 0.001). Hepatomegaly and splenomegaly were less common in MSM group (67 vs. 53% <3 cm, P < 0.001, and 63 vs. 50% <3 cm, P < 0.001, respectively). Poor response to initial treatment with prednisone was recorded in 7.1% of patients with MSM versus 11.5% of those without (P = 0.086). The analysis revealed no independent effect of MSM on event-free survival (EFS), after correcting for differences in EFS related to immunophenotype or initial WBC. CONCLUSIONS: MSM occur mostly in children with BCP ALL who present with less involvement of extramedullary organs, low peripheral blood blasts and white blood cells counts. These findings highlight the importance of including ALL in the differential diagnosis of MSM even in the presence of an apparently normal peripheral blood count. Our study also suggests that MSM are caused by leukemic cells with enhanced biological propensity to remain relatively confined within the intramedullary bone-marrow space. PMID- 19308509 TI - Anatomical study for an update comprehension of clubfoot. Part I: Bones and joints. AB - PURPOSE: The aim of our study was to elucidate the gross anatomical changes of bones and joints in idiopathic clubfeet. METHODS: Gross dissection was carried out on seven idiopathic clubfeet of fetuses aborted between the 25th and 37th week of gestation and compared to two normal feet (27th and 36th week of gestation). Particular attention was paid to the articular surfaces, shapes and angles of all bones and their skeletal relationships. RESULTS: The talar neck trochlea angle in clubfeet ranged from 37 degrees to 41 degrees , in normal feet from 27 degrees to 33 degrees . In clubfeet the deviation of the neck of the talus relative to the body was between 28 degrees and 43 degrees , in normal feet between 22 degrees and 24 degrees . The posterior joint surface was in an anterolateral position and even flat transversely. The head of the clubfeet tali was turned along a longitudinal axis in the opposite direction compared to the normal ones. Instead of a typically saddle-shaped posterior talar surface of the calcaneus, it was triangular and flat transversely, and a bony stability in the subtalar joint was not achieved. The angle of torsion of the calcaneus showed no significant difference between normal and clubfeet. The anterior surface was flat, medially twisted and orientated upwards. CONCLUSIONS: We presume that the calcaneus is the primary fault, which might be explained by pathologic biomechanical forces during development. PMID- 19308513 TI - EPOS/IFPOS Combined Meeting EPOS - 26th Annual Meeting : Sorrento, 11-14 April 2007. PMID- 19308510 TI - Anatomical study for an updated comprehension of clubfoot. Part II: Ligaments, tendons and muscles. AB - PURPOSE: The aim of our study was to evaluate the pathological anatomy of the ligaments, tendons and muscles in clubfeet, and to show whether the dysbalance of shortened and elongated structures is an adaptive process or a primary factor inducing the misshaped bones and cartilagines. METHODS: Surgical exposure was performed on seven idiopathic clubfeet specimens, aborted between the 25th and 37th week of gestation and compared to two normal feet (27th and 36th week of gestation). RESULTS: The medial stabilisation system of the foot was found shortened, but all ligaments could be dissected. On the lateral side, the calcaneofibular ligament in particular was both 'shortened' and 'elongated', depending on the course of the fibres to the axis of motion in the subtalar and talocalcaneonavicular joint. The main difference to the normal feet was found in the thickened tendon of the tibialis posterior forming a bulbus before dividing into fascicules. CONCLUSIONS: We presume the ossification disturbance of the calcaneus to be the primary fault. This disturbance will influence the reduction of the varus position, so ligaments and tendons will be conformed to the misshaped bones. PMID- 19308514 TI - Cell Saver: is it beneficial in scoliosis surgery? AB - STUDY DESIGN: Review of the use of Cell Saver in a non-randomized group of patients undergoing spinal fusion for scoliosis. OBJECTIVES: To determine the efficacy of the use of Cell Saver for spinal fusions for scoliosis. Although Cell Saver is widely used in scoliosis surgery, it is not clear whether its use decreases the need for other transfusions or whether there are certain patients more likely to benefit from its use. The blood collected by Cell Saver intraoperatively is not always of sufficient volume to be returned to the patient, and there are no current guidelines addressing the amount that is likely to be returned to the patient. PURPOSE: The purpose of this study was to determine: Does use of Cell Saver reduce the need for other transfusions in scoliosis surgery? Is there an amount of blood loss at which Cell Saver is likely to be returned? MATERIALS AND METHODS: The study group consisted of 95 children who had undergone posterior spinal fusion between January 2002 and March 2004 with one of two surgeons. One surgeon used Cell Saver, the other surgeon did not. Of the total, 58 patients who underwent posterior spinal fusion with the use of Cell Saver (group 1) were compared with the 37 (group 2) who underwent the same procedure without the use of Cell Saver. RESULTS: Of those in group 1, 34 received Cell Saver blood back as an intraoperative autologous transfusion (IAT) (59%). Patients in the Cell Saver group were just as likely to receive non-Cell Saver transfusions as those in the control group (P = 0.12). Of the 39 patients who lost more than 500 cc of blood, 34 received IAT (87%). No patient who lost less than 500 cc of blood received IAT. CONCLUSION: The use of Cell Saver does not reduce the need for other transfusions in scoliosis surgery. The amount of blood loss at which Cell Saver is likely to be returned is 500 cc. We demonstrated no benefit in the use of Cell Saver in our patient population. PMID- 19308515 TI - Treatment of severe late-onset Perthes' disease with soft tissue release and articulated hip distraction: revisited at skeletal maturity. AB - PURPOSE: Late-onset Perthes' disease is diagnosed after 9 years of age. Conservative treatment and conventional surgical techniques have limited ability to reduce the pressure in the joint or change the shape of the femoral head. We used a combination of soft tissue release and joint distraction with a hinged mono-lateral external fixator for these patients. Ten of our patients reached skeletal maturity and were evaluated. METHODS: Clinical assessment included: Harris hip score, hip range-of-motion (ROM), limb length discrepancy, and the Oxford hip questionnaire for pain and function. Radiographic assessment included: Sharp transverse acetabular inclination, the uncoverage percentage, the epiphyseal index before surgery (modified Eyre-Brook), at frame removal, and, at last follow-up, the epiphyseal quotient (of Sjovall) and the Stulberg classification. RESULTS: Our study included eight boys and two girls (mean age at surgery 12.3 years, range 9.4-15.1, mean age at last follow-up 18.1 years, range 15.2-22.8). The mean follow-up was 5.7 years (range 4.3-7.8). The mean Harris hip score was 86.3/100 (range 48.5-96); one patient had <85 points. The hip ROM was slightly limited in most patients, and seven patients had limb shortening between 1-4 cm. The mean Oxford hip questionnaire score was 17.4/60 (range 12-31). The mean Sharp transverse acetabular inclination of the affected side was 42 degrees (range 36-54) compared to 39 degrees for the unaffected side (P = 0.045). The mean uncoverage percentage was 37% (range 27-47) compared to 20% for the unaffected side (P = 0.017). The mean epiphyseal index was 0.71 (range 0.31-0.92) before surgery, 0.79 (range 0.50-0.93) at frame removal (P = 0.012), and 0.72 (range 0.51-0.89) at last follow-up (P = 0.646). The epiphyseal quotient for the eight unilateral cases was 0.72 (range 0.49-0.91), and the Stulberg classification was type III for three cases and type IV for seven. CONCLUSION: Patient satisfaction for function and pain following the combined procedure was good. Radiographic parameters did not change significantly. This should be regarded as a salvage procedure. PMID- 19308516 TI - Surgical treatment of missed Monteggia lesions in children. AB - PURPOSE: The treatment of an unrecognized Monteggia lesion continues to pose a therapeutic challenge, as evidenced by the variety of surgical techniques described. Moreover, there are high complication and redislocation rates following surgery. This report concerns a surgical technique to reduce a chronic dislocation of the radial head utilizing an ulnar osteotomy and internal fixation. METHODS: Six consecutive cases of missed Monteggia lesions were treated in our institution between August 2001 and September 2003. Patient mean age was 6.5 (range 4-8) years, and the mean interval between injury and surgical procedure was 17 (range 1-49) months. Surgery consisted of an ulnar osteotomy with angulation and lengthening, bone grafting at the osteotomy site, and internal fixation. Open reduction of the radial head, repair or reconstruction of the annular ligament or temporary fixation of the radial head with a transarticular wire was not undertaken. Cast immobilization with the forearm in neutral rotation was maintained for 2 weeks. RESULTS: There was one case of nonunion. At an average follow-up of 3 (range 1.5-4.4) years, all patients had regained painless function of the forearm, good range of elbow and forearm motion, and maintenance of the radial head reduction. CONCLUSIONS: Both angulation and elongation of the ulna are required to allow for reduction of the radial head. We do not see any indication for procedures directed at the radio capitellar joint. PMID- 19308517 TI - Proximal femoral neck penetration after flexible intramedullary nailing for pediatric femur fractures: a rare complication. AB - Flexible intramedullary nailing is a common method used to treat pediatric femur fractures. Complication rates can be high, but most complications are considered minor and do not result in additional unplanned surgeries. Proximal nail penetration is a complication that has only been described once before. The penetration may be asymptomatic, but is still best to be avoided. Two cases of proximal nail penetration of the femoral neck region during nail insertion are presented. A review of the literature with regard to complications and suggestions for avoidance of proximal nail penetration is offered. PMID- 19308518 TI - The anteromedial approach to the psoas tendon in patients with cerebral palsy. AB - PURPOSE: Release of the psoas tendon for flexion deformity of the hip in children with cerebral palsy has traditionally been performed at the pelvic brim, lateral to the neurovascular bundle, or at its insertion into the lesser trochanter. As the psoas tendon is lateral to the pectineus, the traditional exposure of the tendon through an approach medial to the pectineus is limited by the extent to which the pectineus can be retracted proximally. TECHNICAL NOTE: We describe the use of the anteromedial approach used for the developmentally dislocated hip to expose the psoas tendon between the pectineus and the neurovascular bundle. This provides a much better visualisation of the tendon as it crosses the superior pubic ramus to its insertion. The use of this approach has not been described in cerebral palsy. PMID- 19308519 TI - The 1st international congress on early onset scoliosis and growing spine, november 2-3, 2007, madrid, Spain : chairman: behrooz a. Akbarnia, m.d. PMID- 19308520 TI - Intramedullary Kirschner wire (K-wire) fixation of femoral fracture in children. AB - PURPOSE: The aim of this study was to evaluate the efficacy of standard intramedullary Kirschner wires (K-wires) for the treatment of femoral shaft fracture in children. METHODS: We report the results of intramedullary K-wires nailing in 178 children with a mean age of 7.7 years (range, 4-14 years) from 2000 to 2005, retrospectively. A total of 184 diaphyseal femoral fractures were treated with both antegrade and retrograde nailing using the same principles of elastic stable intramedullary nailing (ESIN). The patients were followed for 12 months on average (range, 6-24 months). RESULTS: No major complication (limb length discrepancy >15 mm, non-union, avascular necrosis, knee joint stiffness) occurred during the observation period. All fractures healed within 7.1 weeks on average (range, 5-12 weeks). Associated injuries were seen in 16.9% of the cases. All but seven fractures were reduced by closed manipulation. Early mobilization and weight bearing was allowed. Intramedullary K-wires were removed after an average of 4.8 months (range, 3-12 months) without any complications. CONCLUSIONS: In children, intramedullary fixation by using standard K-wires provides effective treatment for the diaphyseal femoral fracture that has excellent clinical results. Each intramedullary K-wire costs US $5, which adds a cost effective advantage to this method of treatment. PMID- 19308521 TI - Titanium elastic nails for pediatric tibial shaft fractures. AB - PURPOSE: The purpose of our study was to investigate the safety and efficacy of elastic stable intramedullary nailing for unstable pediatric tibial shaft fractures using titanium elastic nails (TENs). To our knowledge, this is the largest series reported in the literature of this specific fixation technique. METHODS: We reviewed all children with tibial shaft fractures treated operatively at our tertiary care children's hospital to find those patients who underwent fixation with TENs. Between 1998 and 2005, we identified 19 consecutive patients who satisfied inclusion criteria. The average age of the patients in our series was 12.2 years (range 7.2-16 years), and mean follow-up was 15.7 months (range 6 28 months). Patient charts and radiographs were retrospectively reviewed to gather the clinical data. Outcomes were classified as excellent, satisfactory, or poor according to the Flynn classification for flexible nail fixation. RESULTS: All patients achieved complete healing at a mean of 11.0 weeks (range 6-18 weeks). At final follow-up, mean angulation was 2 degrees (range 0 degrees -6 degrees ) in the sagittal plane and 3 degrees in the coronal plane (range 0 degrees -9 degrees ). Five patients (26%) complained of irritation at the nail entry site; there were no leg length discrepancies or physeal arrests as a result of treatment. Two patients required remanipulation after the index procedure to maintain adequate alignment. According to the Flynn classification, we had 12 excellent, six satisfactory, and one poor result. CONCLUSION: Although the indications for operative fixation of pediatric tibial shaft fractures are rare, occasionally surgical treatment is warranted. Based on our results, elastic stable intramedullary nailing with titanium elastic nails is an effective surgical technique which allows rapid healing of tibial shaft fractures with an acceptable rate of complications. PMID- 19308522 TI - Pseudoparalysis in the Pavlik Harness: beware of septic arthritis. AB - PURPOSE: Pseudoparalysis of the leg occurring during use of the Pavlik Harness (PH) is a seldom-reported condition. Three cases of pseudoparalysis are reported here to illustrate the need for careful assessment of this condition. METHODS: A series of patients using the PH and presenting with pseudoparalysis was compiled following a retrospective review of case notes. RESULTS: In 3 cases, infants treated for developmental dysplasia of the hip using the PH were found to have developed a pseudoparalysis of the affected limb. All presented with similar signs, with the infant being systemically irritable and reluctant to move the limb. In 2 of the cases, this was related to the harness-its removal resulted in recovery of the affected limb. The third case was proven to be septic arthritis, unrelated to the harness, and treatment by means of arthrotomy, intravenous antibiotics and use of the PH for hip stabilisation was used successfully. In the first two cases, the pseudoparalysis was noticed at routine review clinic, and clinical assessment, ultrasound scan and blood tests were performed. The case of septic arthritis was similarly assessed, following referral by paediatric physicians. CONCLUSION: Pseudoparalysis in the PH is a rare event and requires careful investigation. PMID- 19308523 TI - Is varus osteotomy necessary in one-stage treatment of developmental dislocation of the hip in older children? AB - PURPOSE: Innominate osteotomy, varisation-derotation osteotomy, and shortening osteotomy are the most common surgical procedures used to achieve pelvifemoral realignment in the treatment of children over three years of age with developmental hip dislocation. It is well known that varus osteotomy can improve the acetabular index but it does have some disadvantages. The aim of this study was to discuss the results obtained with patients treated only with Salter osteotomy, derotation osteotomy, and shortening, without varus osteotomy, and to evaluate the need for varisation on the basis of these results. METHODS: Between 1996 and 2001, twenty-four hips, sixteen unilateral and four bilateral, of a total of twenty patients treated with this method and available for final controls were evaluated. The study included sixteen female and four male patients with a mean age of 4 years 2 months (ranging between 2 years 10 months and 8 years). The mean follow-up period was determined as 6.57 years (range 5-10 years). McKay's clinical criteria, Sever's radiological criteria, and the evaluation system modified by Trevor et al. were used for evaluation of the results. RESULTS: Based on McKay's clinical criteria, eighteen hips (75%) were classified as type I, four hips (16%) as type II, and two hips (9%) as type III. According to Sever's radiological criteria, thirteen hips (54%) were graded as grade I, nine hips (37.5%) as grade II, and two (8.5%) as grade III. According to the modified evaluation system of Trevor et al. the results were categorized as excellent in thirteen hips, good in nine hips, and moderate in two hips. Pre operative mean acetabular index was measured as 37.3 degrees (28 degrees -50 degrees ) and early post-operative mean acetabular index as 26 degrees (18 degrees -38 degrees ). In the final radiological examination the mean acetabular index was measured as 18.3 degrees and the mean CE angle as 30.1 degrees (15 degrees -38 degrees ). Avascular necrosis affecting the results developed in five hips. CONCLUSION: It is concluded that in older children with developmental dislocation of the hip (DDH) treated with one-stage combined surgical intervention, adequately stable concentric reduction can be achieved without varisation and that varus osteotomy is not always necessary. PMID- 19308524 TI - Abduction pillow immobilization following hip surgery: a welcome alternative for selected patients. AB - BACKGROUND: Studies have shown that when using a spica cast following hip surgery in children , there are associated complications such as soiling, pressure sores, and osteopenic fractures. METHODS: Charts were retrospectively reviewed for all patients who had undergone hip surgeries by one surgeon at one institution between 1999 and 2005. The method of post-operative immobilization (spica cast or abduction pillow) and complications were recorded for each patient. RESULTS: There was a total of 32 patients. Of these, 11-all of whom had a diagnosis of developmental dysplasia of the hip-were treated using a spica cast. In the 21 patients treated in an abduction pillow, 4 (19%) were switched to a spica cast for the following reasons: (1) distal femoral fracture as a result of vigorous resuscitation during a choking episode, (2) parental preference, (3) for added immobilization over concern of implant loss of fixation, and (4) concern of migration of the hip, which again migrated after being changed to a spica cast and eventually required a pelvic osteotomy. CONCLUSION: It is our opinion that only one of these complications in the children treated with an abduction pillow (the case of possible implant migration) may have been avoided if the child had originally been placed in a spica cast. The child who was successfully resuscitated after a choking incident weeks after the surgery may have had a worse outcome if it had been in a spica cast. This incident occurred at home and the abduction pillow was easily and quickly removed to allow resuscitation. An abduction pillow is a welcome alternative to a spica cast following hip surgery for many patients with neuromuscular and syndromic conditions. However, in children with osteopenic bone at risk for implant failure and in children with unstable hip reductions, a spica cast may be preferable. PMID- 19308525 TI - Reliability of pediobarographs for paediatric foot deformity. AB - PURPOSE: Dynamic pediobarograph measures foot-floor contact pressure during walking and provides a quantitative functional assessment. The goal of this study was to assess the reliability of pediobarograph measurements in normal children and in those with cerebral palsy (CP). MATERIALS AND METHODS: During the first investigation, five non-disabled children and four with CP had three pediobarograph measurements taken of each foot, repeated five times. The pediobarographs were analysed by dividing the foot into five segments; the heel, the lateral midfoot, the medial midfoot, the lateral forefoot and the medial forefoot. A measure of valgus/varus foot posture was defined as the relative medial-lateral difference of combined mid- and forefoot impulse, named valgus/varus index. During the second investigation, 50 children (100 feet) with spastic diplegic CP were studied to calculate the standard error of measurements (SEM), to investigate the number of pediobarograph measurements necessary to obtain accurate results. The third investigation was an inter- and intraobserver study performed on one normal subject's repeated measurements. RESULTS: In the first investigation, the group with CP had a significantly increased variability in the medial midfoot (P = 0.013). The variability of the valgus/varus index had a standard deviation of 13%, demonstrating that this measure is relatively stable. The SEM and 95% confidence interval from the second experiment showed that, even if the accuracy increased with the number of measurements, the greatest gain seemed to be contributed by increasing the number of measurements from 3 to 6. The inter- and intraobserver study showed good to mostly excellent agreement. CONCLUSION: Pediobarograph measurements can be used to monitor and quantitatively assess the progressive changes of foot deformity over time. Pediobarograph is a reliable measurement that shows little variability between measurements at the same occasion and between measurements on different days. Three to six measurements seems practical and adequate to obtain. The technical aspect of measuring shows good repeatability and agreement between observers. PMID- 19308526 TI - Characteristics of patients with congenital clasped thumb: a prospective study of 40 patients with the results of treatment. AB - PURPOSE: Congenital clasped thumb is a deformity that is associated with heterogeneous congenital anomalies and it has been addressed in many congenital syndromes. The aim of this study was to diagnose and evaluate cases of clasped thumb as regards the associated congenital anomalies and syndromes, and evaluation of the results of treatment of such cases. METHODS: A prospective study on 40 patients with 73 clasped thumbs was done. All the patients' data regarding their personal, family, pregnancy and developmental histories were recorded. All the patients were exposed to thorough clinical and radiological examination and genetic assessment. The cases were classified using the Tsuyuguchi et al. (J Hand Surg [Am] 10:613-618, 1985) classification into three types. Conservative treatment was adopted in ten hands, and surgical treatment was performed for 28 hands in 17 patients, with an average follow-up of 26 months. RESULTS: Positive consanguinity was recorded in 57.5% of cases. Associated anomalies were recorded in 77.5% of cases. Type I was the most common one, followed by type III and then type II. Conservative treatment is effective in type I cases when presented early, and all patients were satisfied with the results of surgical treatment. CONCLUSIONS: We reported associated anomalies which are to our knowledge have not mentioned before in the literature which include; congenital blindness, radial deviation of the index finger and ventricular septal defect. We found that 68% of the patients had associated syndromes, and this has not been mentioned before. In this study, we found that there were no difference between type II and type III clasped thumb as regards the pathological findings, severity, the operative procedures, the treatment protocol and the operative results. Properly planned treatment gives satisfactory results. PMID- 19308527 TI - The effect of community-prescribed ankle-foot orthoses on gait parameters in children with spastic cerebral palsy. AB - PURPOSE: To evaluate the efficacy of ankle foot orthoses (AFOs) prescribed in the community for children with cerebral palsy (CP). METHODS: Fifty-six children (32 boys and 24 girls, mean age 8.9 years, range 4-17) who were diagnosed as having CP were enrolled. They were grouped according to the type of CP, diplegic (n = 38) and hemiplegic (n = 18). Three-dimensional gait analyses while patient were barefoot and with AFOs were obtained and analyzed. RESULTS: The spatio-temporal findings were the most significantly changed as a result of AFO use. In the hemiplegic group, stride length was 11.7% (p = 0.001) longer with AFOs in both affected (10.2%) and non-affected (12.4%) legs, and cadence was reduced by 9.7%; walking speed was not affected. In the diplegic group, stride length with AFOs was 17.4% longer compared to barefoot (p < 0.001) and walking velocity improved by 17.8% (p < 0.001); cadence was unchanged. AFOs also increased ankle dorsiflexion at initial contact in both groups. In the hemiplegic group, AFOs produced an average 9.4 degrees increase of dorsiflexion at initial contact (IC) on the affected side (p < 0.001) and 5.87 degrees on the unaffected side (p = 0.007), and an increase of 9.9 degrees (p < 0.001) dorsiflexion at swing, on the affected side. In the diplegic group, dorsiflexion at IC was increased by 13.4 degrees on the right side and 7.8 degrees on the left side (p = 0.05; p > 0.001, respectively) and an increase of 6 degrees (p = 0.005) at swing. In the hemiplegic group of patients, knee flexion at initial contact on the affected side was reduced by 8.5 degrees (p = 0.032) while in the diplegic group we found no influence. The number of patients that reached symmetry at initial double support tripled (from 5.6 to 16.7%) with the use of AFOs. CONCLUSIONS: Our results showed that the use of AFOs improves spatio-temporal gait parameters and gait stability in children with spastic cerebral palsy. It has a lesser effect on proximal joint kinematics. Children with spastic hemiplegia display greater improvement than those with spastic diplegia. PMID- 19308528 TI - Sedation protocols for Ponseti clubfoot Achilles tenotomy. AB - PURPOSE: Ponseti clubfoot management requires percutaneous tenotomy in 90% of cases, typically with local anesthesia. We report two light sedation protocols for outpatient tenotomy. METHODS: Operating room protocol: 24 patients (36 club feet; mean age at tenotomy, 70 days) underwent mask induction with oxygen/nitrous oxide. Pediatric intensive care unit protocol: five patients (eight club feet; mean age at tenotomy, 119 days) underwent intravenous propofol infusion with supplemental oxygen. RESULTS: All patients were discharged several hours after surgery with no complications. Anesthesia that is administered intravenously might have less risk of bronchial reaction than anesthesia that is administered by inhalation. CONCLUSIONS: Our light sedation protocols offer safe alternatives to general anesthesia. Disadvantages include increased cost when compared with local anesthesia. Light sedation can be used effectively and has advantages when treating older infants who might struggle while under local anesthesia. PMID- 19308529 TI - Analysis of halo-orthoses application in children less than three years old. AB - PURPOSE: To evaluate the safety and efficacy of halo immobilization in children younger than 3 years. METHODS: All children less than 3 years of age who were placed in a halo-orthosis at a tertiary pediatric center were reviewed to determine the indications, safety, and efficacy of this method. Adequacy of immobilization and correct technique was established considering the chronology of maturation (skull and brain development). The functional outcome was analyzed. RESULTS: Ten patients (<3 years old) were identified from a total of 150 where halo ring/vest immobilization was used. There were six boys and four girls, with an average age of 2 years (range 10 months to 2 years and 10 months). The average time of immobilization in the halo-orthosis was 75 days (range 33-168 days), and the average follow-up time was 5 years and 2 months (range 1-12 years). There were two halo-related problems and four related to the index procedure. Halo related problems included pin-site infection in one patient, and three pins loosening in another. There were four complications associated with the index procedure, all early in the series, including three cases of pseudarthrosis and one patient that had an insufficient decompression of basilar invagination. Two of the three failed fusions occurred at the occipito-cervical junction, which prompted us to develop new techniques for fusion at this level. CONCLUSIONS: Halo orthosis is a relatively safe and reliable device for immobilization of the upper cervical spine in children less than 3 years old. Understanding the developmental anatomy and a limited CT scan are helpful in pin placement. We recommend the use of a multiple pin construct, perpendicular insertion, and precise tightening with good pin-site care and frequent follow-up. The benefits seem to outweigh the risks in this population, considering the mechanical advantages of the halo orthosis. PMID- 19308530 TI - Scoliosis in-brace curve correction and patient preference of CAD/CAM versus plaster molded TLSOs. AB - PURPOSE: CAD/CAM technology is a newer technique for creating spinal orthoses than standard plaster molded methods. To our knowledge there has been only one previous study of CAD/CAM braces. The purpose of our study was to compare patient preference and in-brace correction of Cobb angle between plaster molded thoracolumbosacral orthoses (TLSO) and CAD/CAM designed TLSOs in a series of patients with scoliosis. METHODS: Ten patients with an average initial Cobb angle of 30.8 degrees (range 18 degrees -46 degrees ) had both a plaster molded TLSO and a CAD/CAM TLSO fabricated for them. In each case, the decision to brace was made by the treating surgeon based on curve magnitude and skeletal maturity. After 3 weeks of 23 h a day wear, in-brace correction of the Cobb angle was measured for each brace based on standard PA spine radiographs. After 3 months of use, patients were asked which brace they preferred. RESULTS: For the CAD/CAM brace, the mean curve correction after 3 months was 51% compared to 44% in the plaster molded TLSO cohort. (p = 0.46). Seven out of nine patients preferred the CAD/CAM TLSO over the plaster molded TLSO. There were no brace complications in either group. CONCLUSION: In our matched cohort study, CAD/CAM TLSOs had at least equivalent if not superior correction of the Cobb angle compared to standard plaster molded TLSOs; 78% of our patients preferred the CAD/CAM brace over the standard TLSO. PMID- 19308531 TI - The Scoliosis Research Society-22 questionnaire adapted for adolescent idiopathic scoliosis patients in China: reliability and validity analysis. AB - STUDY DESIGN: Outcome investigation to verify the internal consistency, reproducibility and validity of the adapted Chinese version of the Scoliosis Research Society-22 (SRS-22) questionnaire for measuring health-related quality of life (HRQoL) in children with idiopathic scoliosis. OBJECTIVE: To develop this questionnaire for the outcome measurement in treating Chinese adolescents with idiopathic scoliosis and evaluate its metric qualities. The SRS-22 questionnaire has proven to be a valid instrument for clinical assessment of patients with idiopathic scoliosis and has been successfully translated into Spanish and Turkish. In most developing countries, however, quality of life and psychological health have been poorly described when treating children with idiopathic scoliosis. METHODS: Trans-cultural adaptation of the SRS-22 questionnaire was carried out according to the International Quality of Life Assessment Project guidelines. The final version was approved by a committee of experts. The questionnaire was completed by 86 adolescents with idiopathic scoliosis who had been treated with a brace; this included 11 males and 75 females, aged from 10 to 18 years (mean 13.9 years). Curve magnitude ranged from 25 degrees to 45 degrees (mean 35.6 degrees ). A subgroup of 30 patients completed the questionnaire again in 3 or 4 weeks. RESULTS: Five common factors were acquired from factorial analysis, and the cumulative contribution ratio was 67.66%. The overall alpha coefficient of the questionnaire was 0.88. Coefficients for individual domains were as follows: function/activity, 0.70; pain, 0.80; self-image, 0.80; mental health, 0.88; and satisfaction, 0.81. The questionnaire as a whole had a test retest correlation coefficient of 0.97. Test-retest correlation coefficients for individual domains were as follows: function, 0.85; pain, 0.96; self-image, 0.96; mental health, 0.95; and satisfaction, 0.91. CONCLUSION: The Chinese version of the SRS-22 questionnaire is eligible in terms of reliability and validity, and can be used to measure HRQoL for adolescent idiopathic scoliosis patients in Mainland China. PMID- 19308532 TI - Three-dimensional computed tomography analysis and anteversion study after periacetabular osteotomy of pelvis in children. AB - PURPOSE: Three-dimensional computed tomography (CT) is the method of choice in understanding the morphological changes after periacetabular osteotomy in children. We studied different parameters and compared aspects of operated hip (OH) with non-operated hip (NOH) to define the maneuver that promotes normalization of the hip during repositioning of the acetabulum. METHODS: A total of 22 patients with 25 OHs underwent CT control scans an average of 4 years after surgery. The patients, with a mean age of 6.8 years, had either Legg-Calve Perthes disease (12 cases) or dysplasia (10 cases).The measurements included the anterior and posterior coverage angles of the hip and version of the acetabulum on axial CT views. The 3D reconstructed images measured the inclination of the antero- and postero-lateral lips, the external rotation and the anterior inclination of the acetabulum. RESULTS: The mean anterior coverage angle was 27 degrees for OHs, 31 degrees for NOHs, and 12 degrees versus 10.3 degrees for the posterior coverage angle. Acetabular anteversion was 2 degrees for OHs (6.3 degrees in the dysplastic OHs) and 6 degrees for NOHs. The mean angle of inclination of the antero-lateral lip was 37 degrees for OHs, 47 degrees for NOHs, and the postero-lateral lip inclination was 56 degrees for OHs and 67 degrees for NOHs. Inferior 3D views showed a mean internal acetabular rotation of 1.5 degrees (4.8 degrees in the dysplastic OH), 3 degrees for NOH. The anterior acetabular inclination angle measured with lateral 3D views was 6 degrees for OHs, 11 degrees for NOHs. CONCLUSION: Our analysis demonstrated a mean anteversion of the acetabulum despite normalization of the anterior coverage of the hip, particularly in the dysplastic group, in which the osteotomized fragments had anteversion superior to NOH. The unexpected external rotation used to improve anterior coverage of a coax magna in Legg-Calve-Perthes disease was responsible for the retroversion and the decrease of the posterior coverage. PMID- 19308533 TI - Does smoking affect the quality of bone regenerate in paediatric limb reconstructive surgery? AB - PURPOSE: To bring to the attention of the orthopaedic fraternity that adolescent children smoke and this has an adverse effect on the bone regenerate during limb deformity corrective surgery. METHODS: Retrospective review of patients undergoing limb deformity corrective surgery with a prolonged frame time and bone healing index. Patients operated on between 1993 and 2005 in a single regional specialist paediatric orthopaedic hospital. RESULTS: Seventeen smoking patients (16 adolescent, one aged 9 years), with prolonged bone regenerate consolidation time of more than double the standard time. Bone-healing index (BHI) was increased in both active smokers and passive smokers. CONCLUSIONS: In the older child/adolescent we should consider smoking (active or passive) as a detrimental factor in prolonging their frame times. We should council these patients and their carers to stop smoking at least during the period of their treatment. PMID- 19308534 TI - New anchoring method for tarsal tendon transfers in myelomeningocele patients. AB - We describe a new anchoring method for tarsal tendon transfers in myelomeningocele patients to protect the sole of the foot from pressure sores and skin necrosis and to loosen the tension of the transferred tendon.Tendon transfer procedures were performed in 51 feet (33 patients) with myelomeningocele. We transferred tibialis anterior tendons to the second or third cuneiform in 19 with equinovarus deformities, and transferred tibialis anterior tendons to the calcaneus through the interosseous membrane in 32 with talipes calcaneus. Clinical results were evaluated with the muscle power of transferred tendons using manual muscle testing 6 months after surgery. The muscle test result was classified as good, fair, and poor.After passing the tendon through the bony hole, a 2.0-mm Kirschner wire was inserted from the sole to the tibia through the ankle joint at neutral. (It extended from the sole through the posterior cortex of the tibia.) The remaining part of the wire was bent and formed into a loop shaped like the Greek letter "zeta" (zeta). The thread was then tied to the loop of the wire as tightly as possible. In this way, there was no contact with the sole during anchoring, thus avoiding ulcers. In addition, the transferred tendon could be kept stable because the patient's ankle was fixed by the Kirschner wire.No cases of wound infection or skin necrosis of the sole occurred. In 49 of the 51 cases, transferred tendons were firmly anchored to tarsal bones. Muscle strength was good for 83%, fair for 13%, and poor for 4%. Consequently, 45 feet could obtain plantigrade pattern during their walking with shoe inserts or occasional use of ankle-foot orthoses.Our anchoring method has the advantage of protecting the sole of the foot from pressure sores and skin necrosis, as well as maintaining tension on the transferred tendon until it settles down in an anchor hole. PMID- 19308535 TI - Meeting Highlights, IFPOS-SICOT 2007. PMID- 19308536 TI - Lower-limb growth: how predictable are predictions? AB - PURPOSE: The purpose of this review is to clarify the different methods of predictions for growth of the lower limb and to propose a simplified method to calculate the final limb deficit and the correct timing of epiphysiodesis. BACKGROUND: Lower-limb growth is characterized by four different periods: antenatal growth (exponential); birth to 5 years (rapid growth); 5 years to puberty (stable growth); and puberty, which is the final growth spurt characterized by a rapid acceleration phase lasting 1 year followed by a more gradual deceleration phase lasting 1.5 years. The younger the child, the less precise is the prediction. Repeating measurements can increase the accuracy of predictions and those calculated at the beginning of puberty are the most accurate. The challenge is to reduce the margin of uncertainty. Confrontation of the different parameters-bone age, Tanner signs, annual growth velocity of the standing height, sub-ischial length and sitting height-is the most accurate method. Charts and diagrams are only models and templates. There are many mathematical equations in the literature; we must be able to step back from these rigid calculations because they are a false guarantee. The dynamic of growth needs a flexible approach. There are, however, some rules of thumb that may be helpful for different clinical scenarios. CALCULATION OF LIMB LENGTH DISCREPANCY: For congenital malformations, at birth the limb length discrepancy must be multiplied by 5 to give the final limb length discrepancy. Multiple by 3 at 1 year of age; by 2 at 3 years in girls and 4 years in boys; by 1.5 at 7 years in girls and boys, by 1.2 at 9 years in girls and 11 years in boys and by 1.1 at the onset of puberty (11 years bone age for girls and 13 years bone age for boys). TIMING OF EPIPHYSIODESIS: For the timing of epiphysiodesis, several simple principles must be observed to reduce the margin of error; strict and repeated measurements, rigorous analysis of the data obtained, perfect evaluation of bone age with elbow plus hand radiographs and confirmation with Tanner signs. The decision should always be taken at the beginning of puberty. A simple rule is that, at the beginning of puberty, there is an average of 5 cm growth remaining at the knee. There are four common different scenarios: (1) A 5-cm discrepancy epiphysiodesis of both femur and tibia at the beginning of puberty (11 years bone age girls and 13 years in boys). (2) A 4-cm discrepancy-epiphysiodesis of femur and tibia 6 months after the onset of puberty (11 years 6 months bone age girls, 13 years 6 months bone age boys, tri-radiate cartilage open). (3) A 3-cm discrepancy-epiphysiodesis of femur only at the start of puberty, (skeletal age of 11 years in girls and 13 years in boys). (4) A 2-cm discrepancy-epiphysiodesis of femur only, 1 year after the start of puberty (12 years bone age girls and 14 years in boys). PMID- 19308537 TI - Proximal femoral osteotomies in children using the Richards hip screw: techniques, outcome and subsequent removal. AB - INTRODUCTION: We investigated the clinical and radiological outcome of proximal femoral osteotomies in children using the Richards hip screw. We also describe previously unreported techniques used in the insertion of the screw to aid satisfactory correction of the neck shaft angle and medialisation. METHODS: Seventy-two children undergoing 81 proximal femoral osteotomies under the care of the senior author over a 10-year period were reviewed. RESULTS: For the primary procedure, there was an overall complication rate of 5%. This included two (2.5%) cases requiring re-operation. There were no fractures and no long-term complications. For subsequent removal of the metalwork, the complication rate was 5%, with a re-operation rate of 1%. CONCLUSION: We conclude that the Richards hip screw is a safe, atraumatic and easy-to-use method of fixation for proximal femoral osteotomies in children, with lower complication. PMID- 19308538 TI - Early clinical and radiological outcomes after double osteotomy in patients with late presentation Legg-Calve-Perthes disease. AB - PURPOSE: Legg-Calve-Perthes disease is an idiopathic avascular necrosis of the femoral head. Although many surgical approaches to treat the late presentation of this pathology have been proposed, there are few reports about the early results of the double osteotomy procedure (femoral varus osteotomy combined with Salter innominate osteotomy). The purpose of this study was to describe the early results obtained with the double osteotomy in patients with late presentation of Legg-Calve-Perthes disease. METHODS: Cross-sectional evaluation of ten patients intervened with double osteotomy. There were seven males and three females with a mean age of 9.2 +/- 1.7 years [standard deviation (SD)]. The average post surgical time of evaluation was of 46.5 +/- 26.2 months. RESULTS: Of the ten evaluated patients, four had a Catterall III and six had a Catterall IV disease. According to Herring classification, three patients were Herring B and seven were Herring C. The epiphyseal extrusion average before and after the surgical procedure was 19.3 +/- 12.4 and 12.1 +/- 14.9%, respectively. In accordance with the Ratliff classification and Lloyd Roberts radiological results, the following were the postoperative clinical results: four good, five fair and one poor. Based on the Stulberg classification, there was one patient in class I, five in class II, three in class III and one in class IV. CONCLUSION: The surgical treatment for late Perthes disease with the best expected outcome is still a challenge. According to the resultsreported here, the double osteotomy could be considered as an alternative to treat this entity. PMID- 19308539 TI - Osteochondroplasty of the femoral head in hip reconstruction for type II late sequelae of septic arthritis: a preliminary report. AB - PURPOSE: To suggest different procedures tailored for hip reconstruction in type II late sequelae of septic arthritis. METHODS: The severely deformed subluxated or dislocated femoral head is reshaped in accordance to radiographic and on-table assessment (osteochondroplasty). Sixteen hips in 13 patients (three bilateral) were the subject of this study. They were all affected during incubation in the first few weeks after birth. Age at operation was in the range 2-12 years (average 5.3). The main complaint was instability, stiffness during walking and the inability to sit comfortably, limb length discrepancy, and mild pain on walking. Preoperatively, the range of motion was limited to a certain degree in different directions in all cases. Plain radiography, computed tomography (CT), or multi-slice CT with reconstruction 3D views were of benefit in analyzing the problem preoperatively. Magnetic resonance imaging (MRI) was performed for selected cases after 2 years to test for the viability of the femoral head. SURGICAL TECHNIQUE: A modified approach was used to adequately expose the iliac bone, the hip, and the upper third of the femur. Meticulous dissection to preserve the amalgamated capsule and a well-planned capsulotomy for later adequate capsulorrhaphy is essential. Debridement to clear the acetabulum from intra-articular fibrosis is attempted prior to acetabular reconstruction (Salter, Dega, or triple pelvic osteotomy). Head and neck reconstruction (osteochondroplasty) is performed according to the nominated topography of the deformed head (beard, collared, staghorn, etc.). A carefully planned reshaping in a manner not disturbing the superior weight bearing articulating surface with the acetabulum will allow easy containment in the reconstructed acetabulum. Associated subluxation or dislocation will dictate adequate shorting with femoral cuts inclined in a manner bringing the impinging overgrown greater trochanter down, achieving a near to normal neck shaft angle. RESULTS: According to the criteria proposed by Hunka et al. (Clin Orthop Relat Res 171:30-36, 1982), a satisfactory result is considered when a stable pain-free hip is achieved with flexion arc >70 degrees and flexion contracture <20 degrees . This was true in 13 hips. It appears that better results are achieved in younger children with minimal intra-articular adhesions limiting hip movements, and with less destruction of the articular cartilage. A final improvement in the range of movement should not be expected before 6-12 months. Intensive physiotherapy to improve postoperative stiffness is required. CONCLUSION: The proposed reconstruction procedure for reshaping the deformed femoral head (osteochondroplasty) is a salvage attempt that achieved a more or less mobile painless stable hip joint besides restoring the normal anatomical relationship, should total hip replacement (THR) be needed in the future. PMID- 19308540 TI - Kingella kingae: osteoarticular infections of the sternum in children: a report of six cases. AB - PURPOSE: Kingella kingae is increasingly recognized as a pathogen of osteoarticular infections (OAI) below the age of 2 years. It was reported that bones and joints which are rarely infected by other pathogens were frequently invaded by K. kingae. Based on a series of six cases, we present the typical clinical and paraclinical manifestation of K. kingae infections of the sternum and sterno-manubrial joint. METHODS: A review of the clinical, laboratory, radiological, microbiological, and molecular data of six consecutive children admitted to a paediatric unit for OAI of the sternum was done. RESULTS: Culture alone allowed for the detection of K. kingae as the responsible pathogen in three cases, molecular methods in the three other cases. Clinical and laboratory findings, as well as imaging methods, proved to be useful in the diagnostic process. CONCLUSION: Our findings suggest that infections of the lower sternum and the junction between the manubrium and the xyphoid process are typical, if not pathognomonic, for the organism. A respective diagnostic and therapeutic protocol was established. PMID- 19308541 TI - Association of Ilizarov's technique and intramedullary rodding in the treatment of congenital pseudarthrosis of the tibia. AB - PURPOSE: Many surgical techniques have been published on how to treat congenital pseudoarthrosis of the tibia (CPT). We combined Ilizarov's fixator with intramedullary nailing of the tibia and developed a procedure which combines the advantages of both methods: Ilizarov's high fusion rate with alignment control and the protection against refracture provided by the intramedullary nail. The results of this approach are presented and discussed. MATERIAL AND METHODS: Seven boys and three girls aged 3-14 years (mean age 8 years 2 months) were treated using our combined technique. In six cases, the CPT was associated with neurofibromatosis. Two strategies were adopted: in six cases, a compression was applied on the bone defect, and in four cases, segmental bone transportation was performed before the compression procedure. The final follow-up (1.2-6.6 years) included a clinical and radiological examination. RESULTS: Tibial union was achieved in nine cases without bone grafting. In one case, tibial union still remains uncertain, despite intertibiofibular bone grafting and additional compression procedures. Thirteen overall complications were noted, including three valgus deformity of the ankle. Bone transportation failed to achieve complete limb lengthening in three cases. One deep infection occurred 4 years after removal of the external fixator. The treatment for this included nail removal and antibiotic therapy for 3 months. Despite a permanent protection of orthosis, a refracture occurred 2 years after nail removal, reverting to the initial level of pseudarthrosis. Another surgical attempt using the same method was then performed with a satisfactory result. CONCLUSIONS: The association of Ilizarov's technique and intramedullary nailing achieved and maintained tibial union in nine of ten patients at final follow-up. It also allowed correction of axial deformities and prevented refracture. Despite the short duration of the follow-up and a high rate of complications in our series, this method can be useful in many cases of CPT in which healing has failed to occur despite many previous surgeries. PMID- 19308542 TI - Treatment of severe early onset Blount's disease by an intra-articular and a metaphyseal osteotomy using the Taylor Spatial Frame. AB - PURPOSE: Severe Blount's disease results in a multiplanar deformity of the lower limb. The mechanical axis is disrupted, there is a rotational deformity, and also shortening of the limb. A depression of the medial tibial plateau causes joint incongruity and instability. The purpose of this study is to review the results of treatment addressing all the aspects of the deformity. METHODS: Four patients were treated for severe Blount's disease. The index procedure consisted of a medial tibial plateau elevating osteotomy, a lateral tibial and proximal fibular epiphysiodesis, a proximal tibial metaphyseal osteotomy, and the application of the Taylor Spatial Frame programmed for correction of varus, rotation and shortening, and preemptive lengthening to compensate for growth arrest. RESULTS: Mechanical axis was corrected from a mean 23 degrees (13 degrees -30 degrees ) to 0 degrees in three patients and 6 degrees valgus in 1. Tibias were lengthened by 1-4 cm. At follow-up, the mechanical axis was 0 degrees in two patients and 7 degrees varus in two. The overlengthening was diminishing as planned. CONCLUSIONS: The double osteotomy and Taylor Frame correction was found to be accurate, safe, and effective for multiplanar deformity correction in severe Blount's disease. PMID- 19308543 TI - Multidisciplinary management of clubfeet using the Ponseti method in a district general hospital setting. AB - PURPOSE: Idiopathic congenital talipes equinovarus (CTEV) is a relatively common complex deformity of the foot that can be successfully managed by the Ponseti method. The purpose of this study was to see if the latter can be effectively administered by non-medical specialists outside a specialist or teaching hospital setting. METHOD: Retrospective review of 24 children (39 feet) with idiopathic congenital talipes equinovarus managed by a physiotherapist-led service in a district general hospital. RESULTS: The median Pirani score at presentation was 4.5 (mean 4.2, range 1.5-6). The median Pirani score for feet requiring tenotomy was 6 (4.5-6), whereas feet not requiring tenotomy had a median Pirani score of 2.5 (1.5-5). A total of 18 feet (46%) underwent an Achilles tenotomy. Foot correction was achieved with an average of 3.4 (2-6) cast changes in the non tenotomy group, and an average of 7.5 (5-13) in the tenotomy group. Successful initial correction of the deformity was achieved in 37 (95%) of the feet studied. One patient (2 feet, 5%) failed local conservative management, requiring tertiary referral. Two children (2 feet) have relapsed, requiring further serial casting. No children required open surgical release. Follow-up was for a mean of 31 months (17-50). CONCLUSIONS: Early results suggest that a combined consultant/physiotherapist-delivered Ponseti service can be effectively and successfully administered in a district general hospital. PMID- 19308544 TI - Tibial tuberosity fractures in adolescents. AB - BACKGROUND: Tibial tuberosity fractures in adolescents are uncommon. We retrospectively reviewed all tibial tuberosity fractures in adolescents (10-19) who presented to our level 1 pediatric trauma center over a 7-year period to review fracture morphology, mechanism of injury, fracture management including return to play, as well as complications. Additionally, we present a review of the literature and treatment algorithm. METHODS: We reviewed the clinical charts and radiographs of consecutive patients with tibial tuberosity fractures between 01 January 2000 and 01 January 2007. Data parameters included the following: patients age and gender, involved side, injury classification, co-morbidities, mechanism of injury, treatment, return to activity and complications. Data were extracted and reviewed, and a treatment algorithm is proposed with some additional insights into the epidemiology of the injury. Nineteen patients met the inclusion criteria. RESULTS: There were 19 patients with 20 tibial tuberosity fractures. The mean age was 13.7 years. There were 18 males and 1 female patient. There were nine left-sided injuries and eleven right-sided including one patient with bilateral fractures. Mechanism of injuries included basketball injury (8), running injury (5), football injury (3), fall from a scooter (2), high jump (1) and fall (1). Co-morbidities included three patients with concurrent Osgood Schlatter disease and one with osteogenesis imperfecta. All were treated with ORIF, including arthroscopic-assisted techniques in two cases. Complications included four patients with pre-operative presentation of compartment syndrome all requiring fasciotomy, one post-operative stiffness and one painful hardware requiring removal. Range of motion was started an average of 4.3 weeks post operatively and return to play was an average of 3.9 months post-operatively. CONCLUSION: Although uncommon, tibial tuberosity fractures in adolescents are clinically important injuries. Early recognition and treatment (closed or open as appropriate) gives good results. All the patients in our series had surgical fixation as per different indications that have been elaborated. It is important for clinicians to recognize that compartment syndrome remains a significant concern post-injury and in the perioperative period. Close monitoring and timely intervention is recommended. A simple treatment algorithm is presented for clinicians to help manage these injuries. PMID- 19308545 TI - Pathologic fractures of long bones in Nigerian children. AB - BACKGROUND: Pathologic or spontaneous fractures are recognised causes of morbidity in children. There is a paucity of data on their aetiology and patterns in African children. METHODS: A combined retrospective and prospective study of 47 children aged 15 and below with pathologic fractures of long bones seen from January 2000 to December 2006. The setting was the Orthopaedic and Traumatology department of the Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria. RESULTS: A total of 47 patients with a mean age of 5.3 years +/- SD with pathologic fractures of 50 long bones were seen and treated. The most commonly affected bones were the femur (28%), tibia (28%) and humerus (26%). Chronic osteomyelitis following inadequately treated acute haematogenous osteomyelitis was the commonest cause of pathologic fractures (74.5%). The mean duration of fracture union was 6.8 +/- 2.3 months and the outcome was satisfactory in 91.4% of cases. CONCLUSIONS: The aetiology of pathologic fractures in the African population is different from their causes in Caucasian children. Chronic osteomyelitis following inadequately treated acute haematogenous osteomyelitis continues to plague many children in our environment. PMID- 19308546 TI - Derotational osteotomy of the proximal radius and the distal ulna for congenital radioulnar synostosis. AB - OBJECTIVE: To evaluate the clinical and functional results of a technical procedure in the surgical treatment of congenital radioulnar synostosis in children. MATERIALS AND METHODS: A prospective study had been undertaken from January 1992 to December 2004. Thirty-four patients with congenital radioulnar synostosis that are fixed in pronation were recruited. Congenital radioulnar synostosis was classified for two types according to Tachdjian's criteria. All patients were treated by resection of the proximal radius and the distal ulna to remove a segmental bone of both parts of the forearm. After K-wires are inserted intramedullarly into both bones, the forearm is derotated manually, followed by cast immobilization. RESULTS: There were 34 patients (52 forearms) with congenital radioulnar synostosis, whom the average age at surgery was 6 years and 3 months. There were two types of congenital radioulnar synostosis: Type 1 in six forearms (11.6%) and Type 2 in 46 forearms (88.4%). The preoperative forearm rotation ranged from 65 degrees to 85 degrees pronation. The postoperative forearm rotation angle was corrected from 0 degrees to 30 degrees ; the best end position appears to be 70-100% of pronation. Of the patients, 78.8% had good or excellent results. All patients were operated on without complications; five patients had loss of correction during cast immobilization. Overall, the patient's ability to perform daily activities showed a marked improvement after surgery. CONCLUSION: This method is a simple and safe technique to derotate the forearms of patients with congenital radioulnar synostosis that are fixed in pronation. PMID- 19308547 TI - Intermittent active motion versus immobilization in the treatment of Staphylococcus aureus-induced arthritis in a rabbit model. AB - PURPOSE: This study has evaluated the effects of immobilization versus intermittent active motion on cartilage and on antibiotic efficacy in a rabbit septic arthritis model. METHODS: Rabbits were infected and assigned to one of four groups: group 1, no treatment without immobilization (allowing intermittent active motion); group 2, cast; group 3, oxacillin without immobilization; group 4, oxacillin and cast. Animals were sacrificed 21 days later. Bacterial counts and lateral radiograms were performed. A radiological score was calculated. RESULTS: Immobilization had no effect on oxacillin efficacy and a deleterious effect on the radiological score. CONCLUSION: Intermittent active motion has allowed a better cartilage healing during the treatment of septic arthritis. PMID- 19308548 TI - Response to Uri Givon. Re: Treatment of early septic arthritis of the hip in children: comparison of results of open arthrotomy versus arthroscopic drainage. PMID- 19308549 TI - Re: Treatment of early septic arthritis of the hip in children: comparison of results of open arthrotomy versus arthroscopic drainage. PMID- 19308550 TI - Acts of loving kindness. PMID- 19308551 TI - Evaluation of the ipsilateral knee more than 40 years after successful closed reduction of DDH. AB - AIM OF THE STUDY: We asked whether the ipsilateral knee of patients treated by closed reduction for developmental hip dislocation (DDH) is affected by osteoarthritis (OA) more than 40 years later. MATERIALS AND METHODS: In total, 61 patients (71 hips) with the diagnosis of DDH were included in the study. The average age at the time of closed reduction was 17.9 months. The follow-up after closed reduction averaged 44.4 years. We determined the following parameters on radiographs, taken in the standing position: minimal joint space width in the medial and lateral compartments of the knee, minimal joint space width of the hip joint, radiological signs of osteoarthritis of the knee and the hip joints (Kellgren and Lawrence), knee alignment, acetabular roof obliquity (AC angle), depth of the acetabulum (ACM angle) and center-edge angle (CE angle) according to Wiberg. All radiographs were measured twice manually with ruler and goniometer by two independent investigators. Interobserver and intraobserver reliability (test retest) was calculated for all parameters. RESULTS: We observed that patients with bilateral DDH had significantly more joint space narrowing in the medial compartment of the knee joints than those with unilateral DDH. The length of follow-up correlated with radiological signs of OA in the hip, but not in the knee. CONCLUSION: We found a significant difference in medial compartment joint space narrowing between patients with unilateral DDH and those with bilateral DDH. The Kellgren-Lawrence classification, however, showed no significant differences between uni- and bilateral DDH. The knee alignment was always at the threshold of physiological values. PMID- 19308552 TI - Post therapeutic lower extremity rotational profiles in children with DDH. AB - PURPOSE: The purpose of this study was to investigate post-therapeutic lower extremity rotational profiles in children with developmental dislocation of the hip (DDH), the differences between these values and those of normal children, and the relationship between these differences and clinical and radiological results. METHODS: In 82 lower extremities of 64 patients, the foot-progression angle was measured clinically and the transmalleolar axis angle photographically, and hip rotations and thigh-foot angle were measured both clinically and photographically. The data obtained were compared with Staheli's data for normal children. In addition, clinical and radiographic data were compared within subgroups and with Staheli's data. Student's t-test and one-way ANOVA were used for statistical evaluation. RESULTS: The medial rotation of the hip, the average clinical value was 44.66 masculine, and the photographic value was 42.28 masculine. Lateral rotation of the hip, the average clinical value was 38.01 masculine, and the average photographic value was 37.29 masculine. Thigh-foot angle, his angle was clinically 8.23 masculine and photographically 9.68 masculine. Angle of the transmalleolar axis, the photographic average was 21.59 masculine. Foot-progression angle, the clinical average was 10.70 masculine. It was determined that the lower extremity rotational profiles of children with DDH treated after walking did not differ from those of normal children, but that the internal and external hip rotations of McKay type III and IV patients were below those of normal children and of McKay type I and II patients. CONCLUSION: Lower extremity rotational profiles in children with DDH who received appropriate treatment were the same as those for normal children. PMID- 19308553 TI - Arthrographic indicators for decision making about femoral varus osteotomy in Legg-Calve-Perthes disease. AB - PURPOSE: There is not always a good outcome after a femoral varus osteotomy (FVO) in those with Legg-Calve-Perthes disease (LCPD), even when the severity warrants surgical treatment. The purpose of this study was to find arthrographic indicators for decision making regarding the likely surgical outcome of a FVO. METHODS: We used an image of an abduction position during preoperative arthrography under general anesthesia that simulated the post-operative relationship between the femoral head and the acetabulum. In the image, we defined two indicators of how deeply the deformed epiphysis was contained within the acetabulum: an acetabular head index in abduction and an epiphyseal slip-in index. Finding the contact point between the top of epiphysis and acetabulum was the key for the epiphyseal slip-in index measurement. In 37 patients (38 hips) who underwent FVOs based on our inclusion criteria, these two indices were measured retrospectively and were analyzed for a correlation with surgical outcome. Surgical outcome was evaluated using a combination of three factors: sphericity of the femoral head (Stulberg's classification), acetabular cover (acetabular head index), and the slope of acetabular roof. RESULTS: The outcome was acceptable in 20 hips (52.6%) and unacceptable in18 hips (47.4%). There was a statistically significance difference in epiphyseal slip-in index between the acceptable group (21.9 +/- 2.8%) and the unacceptable group (15.0 +/- 4.4%) (P < 0.0001). An index of 20% or more determined a safe zone for predicting an acceptable outcome with 80% sensitivity, 89% specificity, and a 7.2 likelihood ratio. However, the acetabular head index in abduction showed no such statistical significance. CONCLUSIONS: In this study, we found that the epiphyseal slip-in index was a reliable indicator for predicting the effectiveness of a FVO. It is worth measuring this index when a surgeon is considering a FVO for a patient with severe LCPD. (Level of Evidence Level III.). PMID- 19308554 TI - Adolescent patellofemoral pain: implicating the medial patellofemoral ligament as the main pain generator. AB - PURPOSE: The purpose of this study is to define the clinical presentation of adolescent patellofemoral pain. METHODS: A review was completed of all patients with patellofemoral pain at a children's hospital sports clinic over a 3-year period. RESULTS: One hundred and one patients (91 female) with 136 symptomatic knees were identified. Mean age was 14.4 years. Knee pain was localized to the anteromedial or anterior region of the knee in 96% of patients and was typically produced with running (94%), jumping (92%) and stair use (69%). On physical examination there was usually a non antalgic gait (99%), no patellofemoral crepitation (98%), normal lower extremity angular (84%) and rotational alignment (94%), with no foot malalignment (>97%). The medial patellofemoral ligament (MPFL) was the most palpably tender area of the knee in 98% of patients. During "lateral apprehension" testing, 89% had pain at the MPFL, but not true apprehension. A "J-sign" was present at terminal knee extension in 65%. Mean Q angle was 18.7 degrees . Means of all radiographic measures were within normal ranges. CONCLUSION: The prototypical patient had anterior/anteromedial knee pain of insidious onset during running and jumping. The most consistent physical findings were focal tenderness at the MPFL, positive terminal J-sign, and an elevated Q-angle. Most patients required only nonsurgical treatments, but 18% underwent surgical interventions for persistent pain. PMID- 19308555 TI - Continuous decompression with intramedullary nailing for the treatment of unicameral bone cysts. AB - PURPOSE: To evaluate the efficacy of decompression of unicameral bone cysts (UBCs) of the long bones with intramedullary nailing and to compare responses to treatment according to location. MATERIALS AND METHODS: We evaluated 48 consecutive patients treated between January 1988 and June 2000. Mean age was 10.3 years. Mean follow-up was 9.8 years. Evaluation was performed according to the radiographic criteria of Capanna. RESULTS: UBCs were located in the proximal humerus (n = 24), humeral shaft (n = 2), proximal femur (n = 19), distal tibia (n = 2) and fibula (n = 1). A total of 62.5% presented a pathological fracture. Successful results were observed in 89.5% (26 total healing, 17 healing with residual radiolucent areas), and there were four recurrences and, in one case, no response to treatment. There was more healing in the humerus than in the femur (92.3% versus 84.2%), and more tendency to restitution ad integrum, although the difference was not statistically significant (P = 0.1499). CONCLUSIONS: Intramedullary nailing is a minimally invasive method, which permits early stability and decompresses the cyst allowing healing. Significant differences were not observed among results from different locations. PMID- 19308556 TI - Metabolic and orthopedic management of X-linked vitamin D-resistant hypophosphatemic rickets. AB - PURPOSE: Therapy of vitamin D-resistant hypophosphatemic rickets (VDXLR) consists of oral phosphate and vitamin D supplements. Bone deformities, pain, and small stature can occur even in children with good compliance, requiring surgical correction and bone lengthening. However, only few surgical reports are available. METHODS: Twelve patients (three males) with VDXLR were followed at our institution. Median age at diagnosis was 3 9/12 years (range, birth to 11 10/12) with a follow-up period of 7 8/12 years (1 9/12-30) and age at last follow-up of 13 6/12 years (2-30). Eight patients underwent surgical correction, three of them in combination with bone lengthening. The corrections were performed at the end of growth in three patients. Clinical endpoints were height, leg axis, and pain. RESULTS: Single bilateral surgical correction was performed in six patients; one patient each had three and five corrections. Bone lengthening was performed in three patients. At last follow-up, the height of seven operated patients was within normal range. In addition, leg axis was normalized in six patients with mild genua vara in two. Only one patient complained of intermittent pain. Bone healing was excellent; surgical complications were rare. There was no radiological evidence of degenerative arthropathy. CONCLUSIONS: Medical treatment remains the main pillar of therapy in children with VDXLR. In case of bone deformity, surgery can safely be performed, independent of age or bone maturation. All patients were satisfied with the results of axial corrective surgery and bone lengthening, and in the majority only one corrective intervention was needed. PMID- 19308557 TI - Adductor release and chemodenervation in children with cerebral palsy: a pilot study in 16 children. AB - PURPOSE: A pilot study with short-term outcomes of a combined surgical and medical intervention for management of generalized lower limb spasticity, hip displacement and contractures of adductors in children with bilateral spastic cerebral palsy. METHODS: A prospective cohort study of 16 children (9 boys and 7 girls) aged 2-6 years with bilateral spastic cerebral palsy was performed. At entry, 5 were classified as level III and 11 as level IV, according to the Gross Motor Function Classification System (GMFCS). The intervention consisted of surgical lengthening of adductor longus and gracilis combined with the phenolization of the anterior branch of the obturator nerve, using 1 ml of 6% phenol, applied under direct vision at the time of lengthening of adductor longus. The hamstring and calf muscles were each injected with Botulinum neurotoxin A at a dose of 4 U/kg/muscle. Serial clinical (hip, knee, ankle joint range of motion), radiographic (migration percentage) and functional data-taken from a functional mobility scale (FMS) or GMFCS-were collected at 3, 6, 12 and 24 months post-operatively. RESULTS: There was a significant increase in hip abduction, knee extension (popliteal angle) and ankle dorsiflexion, maintained for 24 months; mean hip migration percentage decreased from 29 to 21% (P < 0.001). Using a validated mobility scale, significant improvements were noted in gross motor function. There were no complications related to the intervention. CONCLUSIONS: The combined surgical-medical intervention resulted in a reduction of spastic hip subluxation and improvements in gross motor function, as determined by the FMS. The combined intervention is, thus, useful as a temporizing measure, before definitive decisions are made considering such interventions as dorsal rhizotomy, intrathecal baclofen and single-event, multilevel surgery. PMID- 19308558 TI - Analysis of calcaneonavicular coalitions using multi-planar three-dimensional computed tomography. AB - PURPOSE: Computed tomography (CT) is the current imaging standard for confirming the diagnosis of a calcaneonavicular coalition. The purpose of this study was to evaluate three-dimensional (3D) multi-planar CT images of calcaneonavicular coalitions and adjacent tarsal relationships, compared to controls, in order to obtain an understanding of the 3D anatomy for classification and as an aid to surgical resection. METHODS: All patients who underwent CT of bilateral feet at our institution to evaluate a suspected calcaneonavicular coalition between 9/2005 and 3/2008 were retrospectively reviewed. Seventy-four feet (37 patients) were included in this analysis. In addition, 12 patients with a presumed normal calcaneonavicular relationship, who underwent CT of bilateral feet for various other diagnoses, were included as a control cohort. Seven measurements were performed on the 3D reconstructions to quantify the shape of the coalition (dorsal and plantar widths of the calcaneal and navicular contributions, coalition depth of the calcaneal and navicular contributions, and coalition length). The coalitions were classified into four types based on their appearance and their relationship to adjacent tarsal bones. RESULTS: 32/37 patients (86%) were noted to have bilateral involvement (69 coalitions). Coalitions were categorized into four types: Type I (forme fruste)-28%, Type II (fibrous)-23%, Type III (cartilaginous)-45% and Type IV (osseous)-4%. The average shape of the coalition was found to be a curved wedge, which was on average 16 mm wide dorsally, 7 mm wide on the plantar surface, 10 mm in length, and 25 mm in depth. CONCLUSIONS: 3D CT reconstructions enabled the classification of the spectrum of calcaneonavicular coalitions. The shape of the cuboid was found to correlate with the extent of ossification. In Type I or II coalitions, the cuboid extended medially plantar to the fibrous connection. In more complete Type III or IV coalitions, the cuboid was "squared off" and remained lateral to the osseous bridge. An understanding of the 3D anatomy is important when diagnosing milder forms of coalitions, and during resection in order to avoid iatrogenic injury to the calcaneus, head of the talus, or cuboid. PMID- 19308559 TI - Use of end caps in elastic stable intramedullary nailing of femoral and tibial unstable fractures in children: preliminary results in 11 fractures. AB - BACKGROUND: Elastic stable intramedullary nailing (ESIN) is used routinely world wide for treating femoral and tibial shaft fractures in children before skeletal maturity. Overall, ESIN allows rapid limb mobilisation with few complications. However, many centres have experienced some complications resulting from secondary fracture displacement, especially in heavy adolescents and in spiroid or complex fractures due to changes in the nail's position, leading to limb shortening, malrotation and malalignment. End caps were designed by the AO expert group to address these complications. PURPOSE: The purpose of this study is to evaluate the efficiency, safety and tolerance of this new device in our first series of ten patients and 11 fractures. METHODS: Prospective follow-up of our first ten patients operated on with end caps. RESULTS: Our results show that there were no serious complications at a mean follow-up of 13.1 months. There was only one 10-mm leg shortening in a patient in whom the end caps could not be properly inserted. There were no infections, no additional pain and no skin irritation. The use of end caps eased the removal of the nails after bone healing, thereby preventing the formation of ossification over the tips of the nails and appeared to protect the skin next to the nail tip. CONCLUSION: These results seem to prove the advantage of using end caps when performing ESIN in lower limb shaft fractures in children. A longer follow-up that includes more patients and a randomised prospective study should be carried out in the future to determine more precisely the indications of this new device. PMID- 19308560 TI - Histomorphometric analysis of an adolescent distal tibial physis prior to growth plate closure. AB - PURPOSE: Our current understanding of the rate and pattern of physeal closure is based on roentgenographic, magnetic resonance imaging, and qualitative histological studies. The purpose of this report is to provide a detailed histomorphometric/stereological analysis of a distal tibial human growth plate in the process of physiological epiphysiodesis. METHODS: A human distal tibial growth plate was sampled in three regions (anterior, central, and posterior), with each region further separated medially, in the middle, and laterally. The regions were assessed for the location and extent of bony bar formation as well as for physeal height. Companion sections from optimally fixed tissue in the distal 100 microm of the hypertrophic zone were analyzed for hypertrophic chondrocytic volumes. RESULTS: Physis closure started in the middle of the central region of the growth plate, with 46% of the volume in this area occupied by trans-physeal bridging bone. The growth plate was also narrowed with the lowest physeal heights evident in the middle of the central and anterior regions of the physis. Disruption of the regular columns of the physis was evident with the cells arranged in clusters with intervening areas of acellularity. The average hypertrophic cell volume was 5,900 microm(3) and did not significantly differ between different areas of the physis. CONCLUSIONS: This is the first characterization of closure in a human distal tibial growth plate via optimum fixation and stereological techniques. The studied physis was during the earliest phases of closure and provides stereological support that the distal tibial physis closes in a central to medial direction. PMID- 19308561 TI - Local and distant effects of isolated calf muscle lengthening in children with cerebral palsy and equinus gait. Lofterod B, Terjesen T. Journal of Children's Orthopaedics 2008;1:55-62. PMID- 19308562 TI - Answer to HK Graham. PMID- 19308563 TI - New anchoring method for tarsal tendon transfers in myelomeningocele patients. PMID- 19308564 TI - Epidemiology, etiology, and genetic aspects of reduction deficiencies of the lower limb. AB - Although the majority of lower limb deficiencies are of sporadic occurrence and of unknown etiology, genetic factors are involved in a significant number, with variable modes of inheritance. A better-informed public is demanding advice concerning cause and recurrence. Careful scrutiny of the medical history and family tree and attention to phenotypic details may help to delineate entities. At times, specific chromosomal tests are important, mainly when there is bilateral or multiorgan involvement or when limb deficiency is associated with developmental delay and/or mental retardation. This paper is intended to refamiliarize the orthopaedic community with basic genetic aspects regulating reduction deficiencies of the lower limbs, and to emphasize on the importance and indications of genetic counseling. PMID- 19308565 TI - Advanced imaging of the cervical spine and spinal cord in 22q11.2 deletion syndrome: age-matched, double-cohort, controlled study. AB - PURPOSE: The 22q11.2 deletion syndrome is a common genetic syndrome with a wide spectrum of abnormalities. We have previously described multiple anomalies of the upper cervical spine in this disorder. The objective of this study was to use advanced imaging to further define the morphology of the cervical spine and spinal cord in the 22q11.2 deletion syndrome, with a comparison to age-matched controls. METHODS: A total of 32 patients with a 22q11.2 deletion underwent advanced imaging (computed tomography/magnetic resonance imaging; CT/MRI) of the cervical spine. In 27 patients, space available for the cord (SAC); the sagittal diameter of the vertebral body, spinal canal, cerebrospinal fluid (CSF), and spinal cord; and the cross sectional area of the spinal canal, CSF, and spinal cord were measured at each cervical level and compared to 29 age-matched controls. Statistical analysis was performed and potential implications were hypothesized. RESULTS: In 22q11.2 patients, advanced imaging identified 40 pathologies not evident on plain radiographs with potential mechanical and/or neurological implications. These patients also had significantly smaller values (P 0.05). Analyses of two subgroups of the APSF group, anterior release via thoracotomy versus VATS and same-day versus staged surgeries, failed to change any of the above findings. CONCLUSION: Based on this financial analysis, there was no statistically significant differences between the APSF and PSF-TPS groups, with equivalent radiographic corrections. PMID- 19308590 TI - Sagittal balance in scoliosis associated with Marfan syndrome: a stereoradiographic three-dimensional analysis. AB - PURPOSE: Marfan syndrome (MFS) is a genetic disease often marked by the presence of scoliosis. There is no three-dimensional analysis of the deformity in the literature. Our aim was to determine what kind of sagittal balance defines scoliosis associated with MFS, namely a flexion deformity, as it is in scoliosis associated with Chiari I or an extension deformity, as in adolescent idiopathic scoliosis (AIS). To address this issue, we compared the presence or absence of a thoracic scoliosis with the presence or absence of a segment in extension in the thoracic spine. METHODS: In our series, 30 patients diagnosed with Marfan syndrome were prospectively included. In each patient, personalized three dimensional reconstruction from T1 to L5 of the spine was made using stereoradiography. The patients were first separated based on the presence or absence of thoracic scoliosis, in order to compare this with the presence or absence of a segment in extension in the thoracic spine. They were then classified into two groups based on the presence or absence of the segment in extension (meaning containing negative values of inter-vertebral sagittal rotation) in the thoracic spine. RESULTS: Among scoliotic patients with a thoracic scoliosis (17 cases), there were 13 (76.5% cases) with a segment in extension in the thoracic spine and 4 with no segment in extension. CONCLUSIONS: Our results showed that scoliosis associated with MFS is somehow original, demonstrating a sagittal balance in extension (as AIS) in about 80% of thoracic curves, but without this characteristic feature in about 20%. PMID- 19308591 TI - Congenital dislocation of the patella. AB - PURPOSE: Congenital dislocation of the patella is permanent and manually irreducible, and it manifests immediately after birth with flexion contracture of the knee, genu valgus, external tibial torsion and foot deformity. We retrospectively reviewed the results of operative treatment of seven knees in six patients with congenital dislocation of the patella. METHODS: The age of the six patients at diagnosis ranged from 8 days to 3.6 years, with an average of 1.3 years, and their age at the time of operation ranged from 0.6 to 3.9 years, with an average of 2.1 years. Serial casting and/or a brace was attempted before surgery in five of seven knees, leading to improvement in the flexion contracture of the knee. All knees were treated operatively in combination with lateral release, medial plication, V-Y lengthening of the quadriceps, medial transfer of the lateral patellar tendon and posterior release of the knee. RESULTS: Although these deformities were noticed at birth in all seven knees, diagnosis was delayed in three knees due to the low suspicion of the disease and invisible patellae on radiographs. Ultrasonography confirmed the diagnosis of dislocation. The patella was centered in the groove of the femoral condyle after surgery in all knees, but subluxation of the knee with flexion was observed in one knee in which the operation was performed at 3.9 years. Genu valgus and external tibial torsion improved after surgery in all knees. The operated knee was mobile in all cases, with less than 10 degrees flexion contracture of the knee. Flexion contracture did not increase in any of the knees. CONCLUSION: Congenital dislocation of the patella should be suspected in every patient with knee flexion contracture, genu valgus, external tibial torsion, foot deformity and delayed walking. Successful results were obtained when the operation was performed in younger children. Other procedures, such as the semitendinosus tenodesis or tendon transfer, might have to be combined to achieve better stability with flexion in older children. PMID- 19308592 TI - Neurogenic hip dislocation in cerebral palsy: quality of life and results after hip reconstruction. AB - BACKGROUND: At birth, no difference exists between the hips of children with spastic cerebral palsy and the hips of other children. The typical pathologic deformities of the hips in children with spastic cerebral palsy develop as the child gets older. One of our goals was to evaluate the change in quality of life of children with cerebral palsy after hip reconstruction. For classification of the children's condition, we assessed sitting and standing abilities and used the Gross Motor Function Classification System. To evaluate the effect of hip dislocation on the spine, we assessed symmetry while the patient was lying down, sitting, and standing. METHODS: The results of surgical reconstruction of hip joints in 54 patients with severe cerebral palsy who had 66 severely subluxated or dislocated hip joints were retrospectively analyzed. The study was based on a research questionnaire with a mean follow-up of 4 years 10 months. The purpose of the questionnaire was to evaluate the pain, functional abilities, symmetry, and independence of the patients by using the modified Barthel index. RESULTS: Pain was fully relieved in 62 and was eased in 35% of preoperatively painful hips. The functional abilities of sitting and standing were not influenced statistically significantly by the operation. An improvement in sitting symmetry was observed in 40% and improvement in lying symmetry was observed in 32% of the patients. The mean center-edge angle improved from -24.9 degrees +/- 39 degrees (mean +/- standard deviation) to 25.9 degrees +/- 14 degrees . The acetabular index improved from 30.2 degrees +/- 8 degrees to 17.1 degrees +/- 7 degrees . CONCLUSIONS: The independence of the patients, based on the modified Barthel index, did not change significantly after surgery. Most improvements in quality of life were observed in those patients who had pain in the hip before the operation as a result of reduction of pain and improved mobility of the hip. PMID- 19308593 TI - A tale of two tibias: a review of treatment options for congenital pseudarthrosis of the tibia. AB - PURPOSE: Two common treatment options for congenital pseudarthrosis of the tibia (CPT) are intramedullary fixation following resection/shortening of the pseudarthrosis site and reconstruction with an Ilizarov external fixator following resection. We present in detail a narrative of two cases with similar degrees of tibial dysplasia associated with NF-1 treated using these different methods and followed to completion. METHODS: Technical issues and details of the treatment methods from case reports are discussed in depth. The eventual profoundly different outcomes are correlated to the technical variations used. RESULTS: Treatment with the Charnley-Williams rodding method and aggressive bone grafting supplemented by rh-BMP2 resulted in a normal functioning limb at maturity, while treatment with first, an ineffective version of IM rodding, followed by two sessions of bone transport using an Ilizarov fixator failed to gain useful union and eventually resulted in amputation. CONCLUSIONS: Technical details, heretofore inadequately reviewed in the literature, are crucial to the success of either of these commonly utilized treatment methods for CPT. PMID- 19308594 TI - Management of distal tibial medial malleolus type-6 physeal fractures. AB - PURPOSE: Type 6 is an open fracture in which part of the physis is missing. It is the least common physeal fracture, but has the highest rate of complications, particularly the formation of a physeal bar. Without preemptive treatment, a physeal bar always forms, producing growth retardation and angular deformity, and excision of these physeal bars has been uniformly unsuccessful. The distal medial malleolus is a common site for the fracture. METHODS: Strategies for the treatment of two varieties of acute medial malleolar type-6 fractures and two types of late deformities following type-6 fracture are given. The acute fractures were treated with either fat or cartilage applied to the exposed physis. The late deformities were treated with corrective iliac bone grafting. RESULTS: The acute fractures were prevented from forming physeal bars and the two late deformities were fully corrected with good outcomes. CONCLUSION: Fat applied to an acute type-6 physeal fracture has a good chance of preventing bar formation. Ankle deformities due to bars can be corrected by means of iliac bone grafting. PMID- 19308595 TI - Annual Meeting, Vereinigung fur Kinderorthopadie (Paediatric Orthopaedic Society of Germany, Austria, Switzerland) Dusseldorf 2007. PMID- 19308596 TI - Genetic regulation of embryological limb development with relation to congenital limb deformity in humans. AB - Over the last 15 years, great improvements in genetic engineering and genetic manipulation strategies have led to significant advances in the understanding of the genetics governing embryological limb development. This field of science continues to develop, and the complex genetic interactions and signalling pathways are still not fully understood. In this review we will discuss the roles of the principle genes involved in the three-dimensional patterning of the developing limb and will discuss how errors in these signalling cascades correlate to congenital limb deformity in humans. This review is aimed at orthopaedic surgeons wishing to understand the principles of congenital limb deformity related to genetic signalling errors. It is by no means a comprehensive study of the molecular genetics governing the complex interactions involved in each step of limb development. There are however many syndromes involving limb deformity for which the molecular causes are unknown. PMID- 19308597 TI - Triplane ankle fracture with deltoid ligament tear and syndesmotic disruption. AB - In patients with immature skeletons, ligamentous injuries rarely accompany ankle fractures. In this article, we report about deltoid ligament tears and syndesmotic disruptions accompanying triplane ankle fractures in two children, and make recommendations as to the evaluation and treatment of children with such injuries. PMID- 19308598 TI - Elastic stable intramedullary nailing for displaced proximal humeral fractures in older children. AB - PURPOSE: To demonstrate the effectiveness of intramedullary fixation of severely displaced proximal humeral physeal fractures in skeletally immature children using the elastic stable intramedullary nail (ESIN). METHODS: Retrospective recruitment of 14 patients aged 10-15-years old with severely displaced proximal humeral physeal fractures between 1999 and 2004 in a single regional specialist paediatric orthopaedic hospital. The fractures were graded using the Neer classification; severe displacement constituted Neer II-IV or displacement >1 cm and angulation >45 degrees . Patients were followed up and assessed using the Disabilities of the Arm, Shoulder and Hand score (DASH) and the Neer Shoulder score. Radiographs were assessed for deformity. Subjective satisfaction was assessed. RESULTS: Fourteen patients with mean follow-up of 30 months (12-66 m) from surgery. All fractures were radiologically united at a median time of eight weeks (7-10 weeks). At follow-up, Neer Shoulder mean score was 96.79 (range 83 100) and DASH mean score 2.26 (0-7.5). Subjectively 71% were very satisfied and 29% were satisfied. CONCLUSIONS: We commend stabilisation using ESIN in the management of the displaced proximal humeral physeal fracture in older children, once reduction of the fracture has been achieved by either closed or open means. ESIN is safe and allows early return to pre-injury function. PMID- 19308599 TI - Delayed surgical treatment of supracondylar humerus fractures in children using a medial approach. AB - PURPOSE: The appropriate management of supracondylar humerus fractures in children is frequently delayed due to various factors, and there is still no agreement on the treatment of choice. The purpose of this study was to evaluate the effects of delayed treatment of displaced supracondylar humerus fractures on the treatment results and complication rate. METHODS: A total of 31 children with supracondylar humerus fractures who had not received adequate treatment for their displaced fractures were included in this study. The conditions leading to delayed treatment mainly included severe swelling or skin problems around the elbow and the health facility problems unique to our district. The mean delaying time was 6 days (range 2-19 days). There were 24 males and 7 females with a mean age of 7 years (range 1-13 years). The fractures were classified as type III-A (n = 15; 48%) and type III-B (n = 16; 52%) according to Gartland. Surgical treatment consisting of sentiment manual reduction through a medial approach and percutaneous cross-pinning was applied to all. No image intensifier was used during the procedures. A long-arm splint was used for post-operative immobilization. RESULTS: The average hospital stay was 2 days (range 1-3 day) and the patients were followed clinically and radiographically for an average of 4 years (range 2-11 years). Pins were removed at the end of the third week post operatively, at which time the range of motion exercises were begun. None of the patients required physical therapy and full functional recovery was achieved within 3 months in 29 (93.5%) patients and within 5 months in the remaining 2 (6.5%). Two (6.5%) pre-operative nerve injuries (1 interosseous, 1 ulnar) were resolved spontaneously within 3 months post-operatively. At the final follow-up, 7 (22.5%) patients had cubitus varus deformity. Except for 2 (6.5%) pin-tract infections, which were resolved by oral antibiotics and pin removal, none of the patients had early or late complications, not even neurological deficit or myositis ossificans. CONCLUSIONS: Delayed presentation of displaced supracondylar humerus fractures in children did not increase complication rates or unsatisfactory results following an open reduction. Medial approach and cross pinning is an effective and reliable treatment method for these fractures. PMID- 19308600 TI - Long-term effect of repeated injections of botulinum toxin in children with cerebral palsy: a prospective study. AB - PURPOSE: To prospectively evaluate long-term effects of repeated botulinum toxin A (BTX-A) injections in children with cerebral palsy (CP). METHODS: Repeated injections of BTX-A were offered to children with CP, according to clinical indications, for a maximum of four injections within a period of two years. Injections were administered into lower extremity muscles of 26 consecutive children (age 3.7 +/- 1.2 years, 16 boys) with hemiplegic or diplegic CP. Clinical assessments before and one month following each injection included a gross motor function measure (GMFM), a modified Ashworth scale, and range-of motion of knee extension and ankle dorsiflexion. RESULTS: Twelve children received two injections, six received three injections, five received one injection, and three received four injections. The most common reason for discontinuing treatment was the need for orthopedic surgery (n = 17). A long-term effect was demonstrated by a significant increase of the GMFM score before the first injection compared with the last injection for each patient (P < 0.0001). There was no comparable change in the muscle tone or range-of-motion. The mean rate of GMFM change during the study was significantly higher than literature norms for CP children (13.2 vs. 5.37 per year, respectively, P < 0.01). The increase of the GMFM score before and one-month after injection (short-term effect) was significantly higher after the first injection than after the last injection (P < 0.05). Similar results were found for the Ashworth scale and popliteal angle. CONCLUSIONS: Botulinum toxin A injections have a long-term effect on gross motor function in children with CP even though the effect on muscle tone is short-term. The effect apparently declines with repeated injections, with most children benefitting from 2 to 3 injections. PMID- 19308601 TI - Treatment of fixed knee flexion deformity and crouch gait using distal femur extension osteotomy in cerebral palsy. AB - PURPOSE: The purpose of this study was to evaluate the results of distal femur extension osteotomy and medial hamstring lengthening in the treatment of fixed knee flexion deformity in patients with spastic diparetic cerebral palsy. METHODS: A retrospective study was done in a group of 12 diparetic cerebral palsy patients. A distal femur extension osteotomy was performed as part of multilevel surgery on lower limbs. The fixed knee flexion deformity was measured during physical examination, whereas hip and knee flexion in the stance phase and anterior pelvic tilt were both analyzed at kinematics. The pre- and post-surgery results were compared and analyzed statistically. A medical record review was done in order to identify the complications. The mean follow-up was 28 months. RESULTS: A significant reduction of fixed knee flexion deformity at physical examination and knee flexion in the stance phase at kinematics was observed, but with no decrease in hip flexion. As a non-desired effect, there was an increase in anterior pelvic tilt after surgical procedures. With regard to complications, a single patient had skin breakdown at a calcaneous area on one side and the recurrence of deformity was seen in 27% of cases. CONCLUSIONS: In this study, in which fixed knee flexion deformity did not exceed 40 degrees before surgery, the distal femur extension osteotomy was effective in increasing knee extension in the stance phase. However, an increase in anterior pelvic tilt, deformity recurrence and necessity for walking aids are possible complications of this procedure. PMID- 19308602 TI - Associations between orthopaedic findings, ambulation and health-related quality of life in children with myelomeningocele. AB - PURPOSE: Modern principles for treatment of patients with myelomeningocele include early closure of the neural tube defect, neurosurgical treatment of hydrocephalus and treatment aimed at minimizing contractures and joint dislocations. The aim is to achieve a better survival rate and a better quality of life (QOL). Better ambulatory function is thought to improve the management of activities of daily living. This study focused on evaluating which factors might affect ambulation, function and health-related QOL in children with myelomeningocele. METHODS: Thirty-eight patients with neurological deficit from myelomeningocele were examined in an unbiased follow-up. This included a physical examination using validated methods for ambulatory function and neuromuscular status, chart reviews and evaluation of radiographs in terms of hip dislocation and spine deformity. The Pediatric Evaluation of Disability Inventory (PEDI) was used to measure mobility, self-care and social function, and the Child Health Questionnaire (CHQ-PF50) was used to measure QOL. RESULTS: Muscle function class, quadriceps strength, spasticity in hip and/or knee joint muscles and hip flexion contracture as well as the ambulatory level all affected functional mobility as well as self-care/PEDI. Patients with hip dislocation, spinal deformity or those who were mentally retarded also had significantly worse functional mobility. Besides being affected by the severity of the neurological lesion, self-care/PEDI was significantly impaired by hip flexion contracture and absence of functional ambulation. General health-related QOL was significantly lower in this patient group than for US norms. Nonambulatory and mentally retarded patients had a significantly lower physical function of their QOL (CHQ). CONCLUSIONS: The severity of the disease, i.e. reduced muscle strength and occurrence of spasticity around hip/knee, affected ambulation, functional mobility and self care. Acquired deformities (hip dislocation and spine deformity) affected functional ambulation only. Patients with reduced functional mobility and self care experienced lower physical QOL. Children with myelomeningocele had significantly reduced QOL compared to healthy individuals. PMID- 19308603 TI - Local and distant effects of isolated calf muscle lengthening in children with cerebral palsy and equinus gait. AB - PURPOSE: The purpose was to assess the local and distant effects of isolated calf muscle lengthening in ambulant children with cerebral palsy. METHODS: The study included fifteen ambulant children with cerebral palsy (nine with diplegia and six with hemiplegia), average age 8.8 years, Gross Motor Function Classification System (GMFCS) level I and II. None of the children had previously undergone orthopaedic surgery, apart from one child who had tendo-achilles lengthening (TAL) nine years earlier. All the children underwent pre and post-operative clinical examination and three-dimensional gait analysis (gait analysis). Twenty calf muscle lengthenings were performed, ten TAL and ten gastrocnemius recessions (GR). RESULTS: Post-operative ankle kinematics showed significant improvements in all parameters. Ankle power during push-off increased, but only significantly after TAL. Only one limb (5%) was over-corrected. Four limbs (20%) were under corrected and one of these limbs remained in mild equinus position in stance. There was one recurrent equinus (5%) during the follow-up period of three years (range: 13-55 months). Distant effects on joints and segments were more marked in diplegia than in hemiplegia. Ten of 17 kinematic parameters distant from the ankle joint improved significant post-operatively when the preoperative values were 1SD below or above the mean of the normal material. There was no significant deterioration in any of the measured parameters. CONCLUSION: The improvement in ankle kinematics and kinetics supported the experience of other studies. The distant effects, which have previously not been evaluated in three planes, showed improvement in several kinematic parameters indicating that additional surgery in selected patients could be abandoned or delayed. PMID- 19308604 TI - The results of Chiari pelvic osteotomy in adolescents with a brief literature review. AB - PURPOSE: Chiari medial displacement osteotomy is a procedure that uses the cancellous bone of the ilium to contain the femoral head and bear weight. It is the most contraversial osteotomy of the hip joint. This study was therefore conducted to determine the results of this osteotomy with mid-term follow-up in children. METHODS: From 1995 to 2004, 20 Chiari pelvic osteotomies (in 18 patients) were performed. There were 15 male and 3 female patients. The average age at operation was 12.6 years and the mean follow-up was 54 months. The operative technique was as described by Chiari. An iliofemoral approach was used without utilizing a traction table. If there was anterior or anterolateral uncoverage of the femoral head, bone graft augmentation was performed. RESULTS: The angle of the osteotomy averaged 12 degrees , with the distance from the acetabulum averaging 3.2 mm. The average displacement was 42%. Of the 20 Chiari osteotomies, 11 were categorized as excellent, 8 as good and one as fair in terms of clinical and radiological results. Student's t test statistics showed improvements in all radiologic parameters of the hip joint (Sharp angle, center edge angle and coverage of the femoral head). Graft resorption was observed in 25% of the patients. CONCLUSION: There are very rare indications of Chiari osteotomy in patients younger than 10 years. Because of the high rate of graft resorption, Chiari osteotomy should be the last treatment option when there is anterior or anterolateral uncoverage of the hip joint. PMID- 19308605 TI - EPOS 27th Annual Meeting : 9-12 April 2008, Warsaw, Poland. PMID- 19308606 TI - Growth modulation in the management of growing spine deformities. AB - The Hueter-Volkmann law explains the physiological response of the growth plate under mechanical loading. This law mainly explains the pathological mechanism for growing long-bone deformities. Vertebral endplates also show a similar response under mechanical loading. Experimental studies have provided information about spinal growth modulation and, now, it is possible to explain the mechanism of the curvature progression. Convex growth arrest is shown to successfully treat deformities of the growing spine and unnecessary growth arrest of the whole spine is prevented. Both anterior and posterior parts of the convexity should be addressed to achieve a satisfactory improvement in the deformity, albeit epiphysiodesis effect cannot be stipulated at all times. Anterior vertebral body stapling without fusion yielded better results with new shape memory alloys and techniques. This method can be used with minimally invasive techniques and has the potential advantage of producing reversible physeal arrest. Instrumented posterior hemiepiphysiodesis seems to be as effective as classical combined anterior and posterior arthrodesis, where it is less invasive and morbid. Convex hemiepiphysiodesis with concave-side distraction through growing rod techniques provide a better control of the curve immediately after surgery. This method has the advantages of posterior instrumented hemiepiphysiodesis, but necessitates additional surgeries. Concave-side rib shortening and/or convex-side lengthening is an experimental method with an indirect effect on spinal growth. To conclude, whatever the cause of the spinal deformity, growth modulation can be used to manage the growing spine deformities with no or shorter segment fusions. PMID- 19308607 TI - One-stage hip reconstruction in late neglected developmental dysplasia of the hip presenting in children above 8 years of age. AB - PURPOSE: To assess the clinical and radiological results of one-stage hip reconstruction for late neglected developmental dysplasia of the hip (DDH) in children above 8 years of age. METHODS: Nineteen hips in 16 patients, 14 females and two males (three being bilateral), were treated by open reduction adequate shortening (up to 5 cm) with derotation, and limited varization if needed, tight capsulorrhaphy, and appropriate pelvic reconstruction (Salter or triple acetabular osteotomy). The average age at operation 10.6 years (range 8-18 years). The period of follow up ranged from 3 to 9 years. A modified approach for the hip joint and upper femur was utilized, allowing better exposure. Pre- and post-operative plain radiography was performed for all cases and at follow up. Computed tomography (CT) or multislice CT with 3D reconstruction were carried out pre-operatively for recent cases and post-operatively for all, and were found to be helpful in providing a panorama of the dysplastic hip and in planning the required surgery and assessing the results. RESULTS: According to the McKay modified criteria, 15 hips (79%) were clinically excellent to good, while four hips (21%) were fair to poor. Radiographically, according to the Severin modified criteria, 16 hips (84%) were excellent to good and three hips (16%) were fair to poor. Limb length discrepancy ranged from 0.5 to 2.1 cm, as measured by CT scanograms. Complications were avascular necrosis (AVN) in an early case due to limited femoral shortening with resultant excessive stress over the femoral head and subluxation in another case. CONCLUSION: A one-stage hip reconstruction for late neglected cases of DDH have achieved excellent results if adequate shortening with derotation is performed, together with appropriate acetabular reconstruction and tight capsulorrhaphy. Varization should not replace part or all of the required shortening, and should be added if required. PMID- 19308608 TI - Conservative versus surgical treatment of late-onset Legg-Calve-Perthes disease: a radiographic comparison at skeletal maturity. AB - BACKGROUND: Late-onset Legg-Calve-Perthes disease (LCPD; i.e., disease onset after 8 years) has been associated with a more aggressive course and poorer long term outcomes. The optimal treatment method of LCPD is still controversial, especially in this late-onset patient population. The purpose of this study was to evaluate the radiographic outcomes at the end of skeletal maturity in a cohort of patients with late-onset LCPD, comparing the results between conservative and surgical treatment. METHODS: We performed a retrospective comparative study of all 371 patients with LCPD seen at a single institution during a 30-year period. Children younger than 9 years of age at disease onset, bilateral cases, and children that had not reached skeletal maturity at last follow-up were excluded. Clinical data was collected from chart review and radiographs were staged according to the Waldenstrom and Catterall classifications. Patients treated conservatively (adductor tenotomy, bracing and physical therapy, or physical therapy alone) were compared to those treated with more extensive surgery (varus femoral osteotomy or Chiari pelvic osteotomy [CPO]). Final radiographic outcomes were based on a modified Stulberg criteria. RESULTS: Forty-three patients with an average age of 10.8 years and a mean follow-up of 10 years were identified. Twenty-one children received conservative treatment (15 underwent adductor tenotomy, followed by abduction cast or brace and physical therapy; six had only physical therapy) and 22 underwent surgery (16 underwent femoral varus osteotomy and six had CPO). Based on the modified Stulberg criteria, there was no significant difference in radiographic outcome between the surgical and the conservative treatment groups, but there were twice as many patients with a poor result in the conservative group. CONCLUSION: Late-onset LCPD presents with an extensive head involvement and shorter disease course. While there was no statistical difference between surgical and conservative treatment, there was a trend toward better radiographic outcomes when varus derotational osteotomy was performed early in the disease process. PMID- 19308609 TI - Is there a relation between hip torsion, coverage and osteoarthritis of the knee? AB - PURPOSE: Biomechanic factors play a role in the pathogenesis of knee osteoarthritis. The aim of the study was to find out whether there is a relation between femoral, acetabular anteversions, anterior, posterior acetabular coverages and primary osteoarthritis of the knee. METHODS: Thirty patients with primary osteoarthritis of the knee and 29 control subjects were enrolled into the study. Femoral anteversion, acetabular anteversion, McKibbin's instability index, anterior acetabular sector and posterior acetabular sector angles were measured using tomographic scanograms. RESULTS: There was no difference between groups for each parameter (P > 0.05). CONCLUSION: This study did not show any relationship between the axial plane changes in the hip joint and primary knee osteoarthritis. PMID- 19308610 TI - Minimal invasive epiphysiodesis using a modified "Canale"-technique for correction of angular deformities and limb leg length discrepancies. AB - PURPOSE: Leg length discrepancy and angular deformities can, in contrast to adults; easily be corrected with epiphysiodesis during growth. Goal of this study was to evaluate our results of a modified Canale technique for definitive epiphysiodesis treating leg length discrepancy and also angular deformities. METHODS: Between 2000 and 2007, 22 patients (11 boys and 11 girls) were subjected to definitive epiphysiodesis. In total 73 hemiepiphysiodesis were performed (26 proximal tibial, 47 distal femoral). RESULTS: All patients could be followed to the end of growth. Mean follow-up was 32.2 month (range 13-76 months). In 20 patients the epiphysiodesis was planed to correct axis and leg length discrepancy. In two patients' contralateral epiphysiodesis was performed to avoid further leg length discrepancy because of closed physis of the shorter affected side. A staged procedure was necessary in nine patients to achieve the best possible correction. No complications were seen such as wound healing, knee-joint contractures after epiphysiodesis of the distal femur and proximal tibia. In two patients three rehemiepiphysiodesis because of not fully closure of the physis had to be done. CONCLUSION: Definitive epiphysiodesis using this modified Canale technique is a safe, minimal invasive method to correct leg length discrepancy and angular deformities if preoperative planning is performed properly. PMID- 19308611 TI - Multiplanar supramalleolar osteotomy in the management of complex rigid foot deformities in children. AB - PURPOSE: Residual midfoot and hindfoot deformities in rigidly deformed feet present a very complicated surgical dilemma. A plantigrade foot is desirous for proper lower extremity mechanics in a child with ambulatory potential. In this group of patients, soft tissue procedures are no longer an appropriate option, and well-recognized hindfoot procedures, such as talectomy, have many disadvantages. This study reviews the results obtained using multiplanar supramalleolar osteotomy as a salvage procedure to correct deformities of the complex rigid foot in children. METHODS: A retrospective review was conducted of 27 multiplanar supramalleolar osteotomies in 18 children. The underlying diagnosis of the patients included seven severely rigid idiopathic clubfeet, five arthrogryposis, two myelodysplasia, one Ellis-van Creveld, one Streeter's, one cerebral palsy, and one severe burn contracture. The average age at surgery was 5.6 years, and follow-up averaged 8 years. A successful outcome was deemed a plantigrade foot on physical exam with follow-up of at least 2 years and no subsequent tibial surgeries. All failures were included regardless of the length of follow-up. RESULTS: A plantigrade attitude of the hindfoot was obtainable at the time of surgery in all cases. Eighteen of the 27 feet had a successful outcome. Nine of 27 (33%) feet had recurrence of the foot deformity requiring additional surgery. Time to recurrence averaged 5.7 years (9 months-13 years). Complications from the surgery included four minor wound healing problems, two delayed unions, and one screw recession, all of which healed without consequences. There was no evidence of nonunion, growth plate closure, infection, or fracture above or through screw holes. CONCLUSION: The multiplanar supramalleolar osteotomy appears to be a reasonable salvage procedure for severely scarred and complex rigid foot deformities and can be reinstituted for failures due to remaining growth. PMID- 19308612 TI - Great toe metatarsophalangeal arthrodesis for hallux valgus deformity in ambulatory adolescents with spastic cerebral palsy. AB - BACKGROUND: Hallux valgus deformity is a common sequel of spastic cerebral palsy. METHODS: Twenty ambulatory patients (24 feet) suffering hallux valgus deformity, with painful forefoot and restricted footwear, secondary to spastic cerebral palsy acquired perinatally, were treated with great toe metatarsophalangeal (MTP) arthrodesis using percutaneous K-wires for fixation. The mean age at the time of surgery was 16.2 years (range 14-18 years). They were retrospectively evaluated for the results after arthrodesis at a mean interval of 3 years and 4 months (range 3-4 years) by physical examination and radiographs. RESULTS: All patients had a stable painless aligned great toe, with <10 degrees valgus, <20 degrees dorsiflexion and neutral rotation after arthrodesis, evidenced by improvement in pain, cosmesis, functional activity, footwear, callosities and hygiene, as well as by significant improvement in the measures of the MTP and the intermetatarsal angles (IMA) by postoperative radiographs. Neither non-union (pseudoarthrosis) nor recurrence of the deformity developed. Complications included superficial wound slough in a single case. Using the modified American Orthopaedic Foot Ankle Society (AOFAS) Hallux Metatarsophalangeal-Interphalangeal Scale, 18 feet (75%) were classified as excellent and six feet (25%) as good. Neither fair nor poor cases were recorded. CONCLUSION: Hallux valgus deformity in adolescents with spastic cerebral palsy is best treated by great toe MTP arthrodesis to improve segmental foot malalignment and dynamic foot deviation. PMID- 19308613 TI - Complications with flexible nailing of femur fractures more than double with child obesity and weight >50 kg. AB - BACKGROUND: Previous studies report that children above the 95th percentile in weight for their age had an increased risk for complications following titanium elastic nailing for femur fractures. The purpose of this study is to examine whether obesity, defined as body mass index (BMI) > 95th percentile, and/or simple weight correlates with an increased rate of complications. METHODS: The incidence of complications was compared between obese and non-obese patients and also between patients who weighed >/=50 kg and those <50 kg. RESULTS: The overall complication rate was 23% (16/71). The complication rate was 17% (10/58) for "non obese" patients and 46% (6/13) for "obese" patients. This difference was statistically significant (P = 0.03). The complication rate was 46% (6/13) in children who weighed >/=50 kg and 17% (10/58) in children who weighed <50 kg. This difference was also statistically significant (P = 0.03). CONCLUSIONS: This study demonstrates that obesity (BMI > 95th percentile) and weight over 50 kg predispose patients to increased risk of surgical complications when undergoing flexible elastic nailing for femur fractures. Both obese children and children weighing >/=50 kg were two times more likely to have a complication when undergoing this procedure. PMID- 19308614 TI - Digital imaging data on CD-R: a time trap for orthopaedic surgeons in outpatient clinics. AB - PURPOSE: X-rays are presented on CD-Rs in a digital format with increasing frequency. This technique is potentially more time-consuming for the clinician compared to conventional pictures. METHODS: Ten sets of six X-rays for each case were prepared in both the conventional way and in digital format on CD-R. The order was randomised. Time in seconds was measured for six experienced orthopaedic residents to present the most recent a.p. view out of a given set. As a precondition, the computer was turned on and the same viewing software was used for all digital sets. The results were compared using a non-linked Student's t test (significance level P = 0.05). RESULTS: The presentation of conventional X rays required 21 s (+/-7.5 s) and of digital X-rays 90 s (+/-27 s), respectively (P < 0.001). CONCLUSION: In spite of ideal conditions, digital X-rays on CD-R need significantly more time in the orthopaedic clinic. In major centres, patients present with different software and software in other languages, which increase the required time even further. This latter problem will be assessed in another study. This increase of preparation time required by a highly qualified staff member has implications on the economics and logistics and has a negative impact on daily clinical practice. One of the solutions might be that the CD-Rs be given to the registration desk and the data being already prepared when the patient sees the doctor. This can also include it being implemented in the digital system used in the inpatient part of the hospital and, therefore, making any other consultations or second opinions easily accessible. PMID- 19308615 TI - Varus producing osteotomy for patients with lateral pillar type B and C Legg Calve-Perthes disease followed to skeletal maturity. Castaneda et al. JCOR (2008), 2:373-379. PMID- 19308616 TI - Response to Professor Ghanem's Letter to Editors on the article entitled: "Varus producing osteotomy for patients with lateral pillar type B and C Legg-Calve Perthes disease followed to skeletal maturity" by Pablo Castaneda et al. PMID- 19308617 TI - Fifth International Clubfoot Congress IFPOS & ICFSG, August 27-28, 2008, Hong Kong. PMID- 19308618 TI - Professor henri bensahel (1928-2009). PMID- 19308619 TI - Arthroscopy of the hip in children and adolescents. AB - Intra-articular abnormalities of the hip, such as labral tears, loose bodies, chondral lesions, ligamentum teres tears and femoral acetabular impingement are increasingly being recognized in the pediatric age group. Evaluation for these abnormalities starts with a good history and physical exam. Radiographic imaging with plain films and magnetic resonance imaging help confirm the clinical impression. Arthroscopy of the hip can be utilized to diagnose and treat these abnormalities. Arthroscopy of the hip is a challenging procedure with a learning curve that requires a thorough knowledge of the anatomy of the hip. The hip is a deeply recessed joint that has a large muscular envelope, thick joint capsule and convex and concave surfaces of the femoral head and acetabulum, respectively. The normal anatomy may be distorted due to childhood developmental disorders such as hip dysplasia, Legg-Calve-Perthes Disease and Slipped Capital Femoral Epiphysis that adds additional challenges to the arthroscopist. Isolated intra-articular abnormalities occur rarely and an underlying morphologic abnormality should be identified which also requires management. Complications can be minimized with attention to detail. PMID- 19308620 TI - Complete obstetric brachial plexus palsy: surgical improvement to recover a functional hand. AB - OBJECTIVE: Complete obstetrical brachial plexus palsy remains a difficult situation for the child and his/her family. The quality of spontaneous reinnervation is rarely good and always leads to a non-sensitive and non functional hand, even if abduction of the shoulder and elbow flexion do spontaneously recover. The aim of this study was to assess the results from nerve reconstruction in cases of complete palsy and to demonstrate the effect of a change in surgical technique on the outcome of hand function. METHODS: Thirty pediatric patients with complete obstetric brachial plexus palsy were operated on in our department between 1987 and 2003. Twenty-five of these patients were clinically reviewed and evaluated by a physiotherapist and a surgeon (not the surgeon who performed the surgery). Functional assessment was based on the Gilbert shoulder score, the Gilbert-Raimondi score for elbow function and the Raimondi hand score. All children underwent a nerve reconstruction as graft and/or intra- or extra-plexual neurotization. Our neural surgical strategy changed between 1995 and 1996 to one that addresses the function of the hand and the wrist. A secondary surgical intervention was required in 18 cases. The most frequent procedures were a radial rotation osteotomy and a tendon transfer restoring wrist extension. RESULTS: Mean follow-up was 7 years and 10 months. Among children operated on with the first surgical strategy-the pre-1995-1996 period-hand function was good in three cases, satisfactory in four cases and unsatisfactory in one case. Among children for whom the second surgical strategy was used-1995-1996 and later-hand function was good or very good in eight cases, satisfactory in four cases and unsatisfactory in two cases. When the 25 patients were assessed for shoulder function, the functional result was good or very good in 12 cases, satisfactory in seven cases and unsatisfactory in six cases. In terms of elbow function, the results were good or very good in 14 cases, satisfactory in eight cases and unsatisfactory in three cases. All hands recovered sensation to a certain degree. CONCLUSION: The surgical results are encouraging in terms of shoulder and elbow function, but not as good for hand function. With the change in neural surgical strategy in 1995-1996, when more focus was placed on the hand (second surgical strategy), the results on hand function improved relative to those obtained with the first surgical strategy. It must also be noted that hand recovery requires more time, which may partially explain why functional results are not as good for the hand as for the shoulder and elbow. These results demonstrate that early surgical exploration is useful in complete obstetrical brachial plexus palsies and that there is a need for neural reconstruction of the lower trunk. PMID- 19308621 TI - Surgical management of windblown hand: results and literature review. AB - PURPOSE: Windblown hand is a congenital anomaly characterized by multiple hand deformities. The condition is extremely rare as shown by the paucity of cases reported in the literature. The deformities, which can result in a cosmetically unsatisfactory appearance and, if left untreated, maladaptive behavior, are progressive in nature. Consequently early treatment is necessary. METHODS: We operated upon 23 hands in 18 patients (age range at surgery 6 months to 16 years) at a tertiary care center over a period of 7 years. The patients were followed for an average period of 7 years. The surgical approach was chosen based on the severity of the condition according to Zancolli and Zancolli's classification [Hand Clin 1:443-456 (1985)]. RESULTS: According to the criteria of Wood and Biondi [J Hand Surg 15A:431-438 (1990)], of the 23 hands operated upon, 17 had excellent cosmetic results, and 15 had excellent functional results. The results were better in patients undergoing early surgery-before the age of 2 years. Relapse of the deformity to a lesser extent than the original condition was seen in two hands at the last follow up. CONCLUSION: Definitive conclusions on this condition cannot be drawn due to limited experience in the surgical management of this rare condition. We believe that early surgical management is probably the best option available for the patient based on the results obtained. Early surgery and good post-operative compliance from patients can facilitate successful management of this rare condition with predictable results. PMID- 19308622 TI - A rare pentad of foot and ankle deformities in hyperlax children. AB - OBJECTIVE: The aim of this paper is to describe the clinical features and radiological appearance of a rare and complex lower leg and foot deformity in hyperlax children. METHODS: Four children were included in the study; of these, two had bilateral feet involvement. All deformities were present since birth, comprising a pentad of conditions: (1) pathologic external tibia and fibula torsion, (2) ball and socket ankle joint with medial subluxation and talus medial subluxation, (3) excessive hindfoot valgus, (4) peroneal and Achilles tendon displacement anterior to the lateral malleolus and (5) generalized ligament laxity. All children had failed cast manipulation. Only two had undergone a minimal soft tissue procedure, but there had been no improvement. RESULTS: All patients were ambulatory and pain free at the mean age of 6.5 years. CONCLUSION: Conservative or minimal soft tissue procedures are not effective in restoring the anatomy of such feet. PMID- 19308623 TI - Surgical treatment of traumatic pediatric humeral diaphyseal fractures with titanium elastic nails. AB - BACKGROUND: Nearly all pediatric humeral shaft fractures can be successfully treated with closed methods. Some patients, however, require internal fixation either because of an inability to maintain an adequate reduction, significant soft tissue injury, or concomitant fractures. METHODS: This is a retrospective review of all traumatic humeral shaft fractures treated at our hospitals between 1999 and 2006. Thirteen pediatric patients ranging in age from 4.8 to 16.7 years (mean age 12.0 years) were treated surgically with titanium elastic nails (TENs). Relative surgical indications included open fractures, inability to maintain an acceptable reduction, the presence of ipsilateral forearm fractures (floating elbow), concomitant lower extremity fractures, and closed head injury. Two patients had associated radial nerve injury at presentation. RESULTS: The patients were followed for a mean of 29 months. All fractures healed in good alignment. There were no intraoperative complications, including neurologic or vascular injury, and no patient developed an infection postoperatively. Two patients had nail migration, one of whom developed nail protrusion through the skin. One patient with preoperative radial nerve injury ultimately underwent tendon transfer to restore wrist extension. Of the 13 patients, 12 reported a full return to sports and other activities with no limitations or discomfort. CONCLUSIONS: When surgical stabilization of pediatric humeral shaft fractures is indicated, TEN fixation is effective and has a high rate of union and a low rate of complications. This technique is familiar to most orthopaedic surgeons treating pediatric fractures. LEVEL OF EVIDENCE: Level of evidence: IV (case series). PMID- 19308624 TI - Femoral shaft fractures treated with stainless steel flexible nails in children aged between 5 and 12 years at the HUSVP: a two-year follow-up. AB - PURPOSE: The purpose of this study is to show our experience with stainless steel flexible nails in the treatment of femoral shaft fractures in children aged between 5 and 12 years and to observe if the type of material used for nails affects the results. MATERIALS: A prospective, descriptive study of a series of cases was performed to evaluate patients treated at the Hospital Universitario San Vicente de Paul (HUSVP) who had been followed up for at least 24 months. RESULTS: The study group was composed of 48 patients, with an average age of 8.6 years. According to the AO classification, the most common type of fracture was 32A3.2 (61%). The average follow-up period was 43 months. The alterations observed were six cases of clinical length discrepancy, two cases of overgrowing, four cases of shortening, and two malunions in valgus. These alterations had no repercussion in the children's function. The follow-up is the main strength of this study, as it demonstrated that, 2 years after surgery. CONCLUSION: The results with stainless steel nails are as good as those with titanium nails with a shorter follow-up as reported in the literature. Given the high costs of osteosynthesis material in our country, the results of this study provide a less expensive alternative. Level of evidence IV, case series. PMID- 19308625 TI - Corrective lower limb osteotomies in children using temporary external fixation and percutaneous locking plates. AB - BACKGROUND: We describe a new method for corrective osteotomy and the fixation of lower limb deformities in children. METHODS: Following osteotomy, the desired position is obtained and temporarily stabilized using an external fixator. Definitive fixation is performed with a plate and locking screws inserted percutaneously under fluoroscopic guidance. The procedure was performed in 18 segments in 11 patients. Corrections were performed in all planes. RESULTS: The mean follow up was 18 months (range 6-36 months). All patients were corrected to within 2 degrees of that which was planned. Union was obtained in 16 segments in ten patients within 6-16 weeks. CONCLUSIONS: The method was found to be effective for the correction of deformity in the pediatric population. Advantages compared to conventional methods include minimal soft tissue dissection and the ability to adjust the position before definitive fixation. PMID- 19308627 TI - EPOS 28th Annual Meeting Lisbon, 1-4 April 2009. PMID- 19308626 TI - The 2nd international congress on early onset scoliosis and growing spine, november 7-8, 2008, montreal, Quebec, chairman: behrooz a. Akbarnia, MD. PMID- 19308628 TI - Bone histomorphometry in male idiopathic osteoporosis. AB - The pathogenesis of male osteoporosis at the cellular level is still elusive. We performed histomorphometric analysis of bone biopsy samples from 51 eugonadal men with idiopathic osteoporosis. Their median age was 54 (range 29-73) years. Eighty two percent of the patients had a fracture history, and 57% had vertebral fractures. Bone volume, trabecular thickness, wall thickness, and osteoid thickness were significantly reduced in osteoporotic men compared with healthy men. Erosion depth was similar, as were the bone remodeling parameters such as bone formation rate, mineral apposition rate, and activation frequency. In the osteoporotic men, osteoid thickness was correlated to bone mineral density at the lumbar spine (R(2) = 0.19, P < 0.01); together with wall thickness, the two parameters could explain 27% of the variation in lumbar spine bone mineral density. The osteoid thickness was correlated to anthropometric variables such as body weight (R(2) = 0.24, P < 0.001) and body mass index (R(2) = 0.14, P < 0.01), as well as to serum estradiol levels (R(2) = 0.14, P < 0.01) and to the ratio insulin-like growth factor-1 (IGF-1) to IGF-binding protein-1 (IGFBP-1) (R(2) = 0.12, P < 0.01). Regression analysis showed that 36% of the variation in osteoid thickness could be predicted by body weight and estradiol levels. In conclusion, bone histomorphometry in male idiopathic osteoporosis was characterized by thin bone structural units, which might suggest osteoblast dysfunction. Bone histomorphometry parameters were associated with low body weight, low estradiol levels, and increased levels of IGFBP-1, supporting the notion that estrogens and IGFs play regulatory roles in male bone turnover. PMID- 19308629 TI - 25-hydroxyvitamin D is not associated with carotid intima-media thickness in older men and women. PMID- 19308630 TI - P2X7 nucleotide receptor plays an important role in callus remodeling during fracture repair. AB - The P2X7 nucleotide receptor (P2X7R) is an ATP-gated ion channel expressed in bone cells. Homozygous null P2X7R (P2X7R(-/-)) mice have reduced bone formation, so we hypothesized that P2X7R(-/-) mice have impaired fracture healing compared to P2X7R(+/+) control mice. To test the hypothesis, adult P2X7R(-/-) mice and P2X7R(+/+) mice were studied. Osteotomy of the right femur was performed and a stainless-steel pin was inserted into the medullary cavity to stabilize the fracture site. No differences in callus development were seen in the radiograph, micro computed tomography, or dual-energy x-ray absorptiometry measurements. Mechanical testing showed that the recovery of ultimate force, stiffness, and energy to failure were slightly decreased in P2X7R(-/-) mice compared with the control. Histomorphometric measurements of the callus revealed that mineralizing surface and bone formation were significantly decreased, by 22% (p < 0.001) and 29% (p < 0.05), respectively, in P2X7R(-/-) mice in comparison with the wild-type control. These data show that a null mutation of the P2X7R does not affect the amount of callus formed in our osteotomy fracture model. However, callus remodeling was significantly delayed. Our data suggest the different role of the P2X7R in woven bone and lamellar bone formation. PMID- 19308631 TI - Bone mineral accrual across growth in a mixed-ethnic group of children: are Asian children disadvantaged from an early age? AB - We investigated the contribution of ethnicity, physical activity, body composition, and calcium intake to bone accrual across 7 years of growth. We assessed 80 Caucasian and 74 Asian boys and 81 Caucasian and 64 Asian girls at baseline and retained 155 children across all 7 years. Ethnicity, physical activity, and calcium intake were assessed by questionnaire; fat mass, lean mass, and bone mineral content (BMC) of the whole body (WB), lumbar spine (LS), total proximal femur (PF(TOT)), and femoral neck (FN) were measured using DXA (Hologic QDR 4500). We aligned children on peak height velocity and utilized multilevel modeling to assess bone mineral accrual. Height and lean mass accounted for 51.8% and 44.1% of BMC accrual in children. There was a significant difference in physical activity, calcium intake, and lean mass between Asians and Caucasian boys and girls at baseline and conclusion (p < 0.05). In boys, physical activity and ethnicity significantly predicted BMC accrual at the FN. In girls, Asians had significantly lower PF(TOT) and FN BMC. Calcium was a significant predictor of WB BMC accrual in boys and girls. In conclusion, our findings highlight the importance of accounting for ethnicity in pediatric studies. Physical activity, dietary calcium, and lean mass positively influence bone accrual and are lower in Asian compared to Caucasian children from a very young age. PMID- 19308632 TI - Correlation between Ka/Ks and Ks is related to substitution model and evolutionary lineage. AB - In 2005, Wyckoff and coworkers described a surprisingly strong correlation between Ka/Ks and Ks in several data sets using the LPB93 algorithm. This finding indicated the possibility of a paradigm shift in the way selection strength can be measured using the Ka/Ks ratio. We carried out a calculation of Ka and Ks using six different algorithms on three cross-species orthologous data sets and found a highly variable correlation among the algorithms and lineages. Algorithms based on the GY-HKY substitution model exhibit a weaker positive correlation or a stronger negative correlation than those based on the K2P and JC69 substitution model. Even if one algorithm shows a positive correlation between Ka/Ks and Ks in a warm-blooded lineage, it may show no correlation in a cold-blooded lineage. This algorithm-related and evolutionary lineage-related correlation indicates the need for great caution in drawing conclusions when using only one Ka and Ks algorithm in a genomewide analysis of selection strength. Our results indicated that currently used algorithms for Ka and Ks calculations are flawed and need improvements. PMID- 19308633 TI - Patterns of variation in the inhibitor of apoptosis 1 gene of Aedes triseriatus, a transovarial vector of La Crosse virus. AB - Aedes triseriatus mosquitoes transovarially transmit (TOT) La Crosse virus (LACV) to their offspring with minimal damage to infected ovaries. Ae. triseriatus inhibitor of apoptosis 1 (AtIAP1) is a candidate gene conditioning the ability to vertically transmit LACV. AtIAP1 was amplified and sequenced in adult mosquitoes reared from field-collected eggs. Sequence analysis showed that AtIAP1 has much higher levels of genetic diversity than genes found in other mosquitoes. Despite this large amount of diversity, strong purifying selection of polymorphisms located in the Baculovirus inhibitor of apoptosis repeat (BIR) domains and, to a lesser extent, in the 5' untranslated region seems to indicate that these portions of AtIAP1 are the most important. These results indicate that the 5'UTR plays an important role in transcription and translation and that the BIR domains are important functional domains in the protein. Single nucleotide polymorphisms (SNPs) were compared between LACV-positive and -negative mosquitoes to test for associations between segregating sites and the ability to be transovarially infected with LACV. Initial results indicated that five SNPs were associated with TOT of LACV; however, these results were not replicable with larger sample sizes. PMID- 19308634 TI - Evolution of the metazoan-specific importin alpha gene family. AB - Importin alphas are import receptors for nuclear localization signal-containing proteins. Most animal importin alphas assort into alpha1, alpha2, and alpha3 groups. Studies in Drosophila melanogaster, Caenorhabditis elegans, and mouse suggest that the animal importin alpha gene family evolved from ancestral plant like genes to serve paralog-specific roles in gametogenesis. To explore this hypothesis we extended the phylogenetic analysis of the importin alpha gene family to nonbilateral animals and investigated whether animal-like genes occur in premetazoan taxa. Maximum likelihood analysis suggests that animal-like importin alpha genes occur in the Choanoflaggelate Monosiga brevicollis and the amoebozoan Dictyostelium; however, both of these results are caused by long branch attraction effects. The absence of animal-like alpha genes in premetazoan taxa is consistent with the hypothesis that they duplicated and then specialized to function in animal gametogenesis. The gene structures of the importin alphas provide insight into how the animal importin alpha gene family may have evolved from the most likely ancestral gene. Interestingly, animal alpha1s are more similar to plant and fungal alpha1-like sequences than they are to animal alpha2s or alpha3s. We show that animal alpha1 genes share most of their introns with plant alpha1-like genes, and alpha2s and alpha3s share many more intron positions with each other than with the alpha1s. Together, phylogenetics and gene structure analysis suggests a parsimonious path for the evolution of the mammalian importin alpha gene family from an ancestral alpha1-like progenitor. Finally, these results establish a rational basis for a unified nomenclature of the importin alpha gene family. PMID- 19308635 TI - Phylogenetic inference with weighted codon evolutionary distances. AB - We develop a new approach to estimate a matrix of pairwise evolutionary distances from a codon-based alignment based on a codon evolutionary model. The method first computes a standard distance matrix for each of the three codon positions. Then these three distance matrices are weighted according to an estimate of the global evolutionary rate of each codon position and averaged into a unique distance matrix. Using a large set of both real and simulated codon-based alignments of nucleotide sequences, we show that this approach leads to distance matrices that have a significantly better treelikeness compared to those obtained by standard nucleotide evolutionary distances. We also propose an alternative weighting to eliminate the part of the noise often associated with some codon positions, particularly the third position, which is known to induce a fast evolutionary rate. Simulation results show that fast distance-based tree reconstruction algorithms on distance matrices based on this codon position weighting can lead to phylogenetic trees that are at least as accurate as, if not better, than those inferred by maximum likelihood. Finally, a well-known multigene dataset composed of eight yeast species and 106 codon-based alignments is reanalyzed and shows that our codon evolutionary distances allow building a phylogenetic tree which is similar to those obtained by non-distance-based methods (e.g., maximum parsimony and maximum likelihood) and also significantly improved compared to standard nucleotide evolutionary distance estimates. PMID- 19308636 TI - Identification and characterization of freshwater algae from a pollution gradient using rbcL sequencing and toxicity testing. AB - One approach in toxicity testing using microalgae is to assess the modulation of light energy absorbed as a result of exposure to contaminants. In this study, four strains of microalgae were isolated to obtain a variety of taxa for testing from sites receiving various levels of environmental stressors around Christchurch, New Zealand. The strains were characterized by partially sequencing rbcL, a routinely used gene in plant phylogenetics with a large existing database of strains. Based on morphological observation and gene sequences, the strains were identified as Chlorella sp., Neochloris sp., and Choricystis minor. The isolates were exposed to the herbicide glyphosate and the metal zinc, and their responses were measured using the ToxY-PAM system. Chlorella sp. was the most sensitive. Two strains of Choricystis minor were isolated from different ponds in an effluent gradient at a sewage treatment plant. Analysis of variance indicated that the isolate from the least contaminated pond was more sensitive to zinc (although regression analysis did not show this result). This suggests that the selective pressure exerted on algal strains by a contamination gradient over short a distance is detectable by both genetic and physiological methods, with implications for sourcing appropriate indicator organisms from the environment. PMID- 19308637 TI - Perchlorate exposure induces hypothyroidism and affects thyroid-responsive genes in liver but not brain of quail chicks. AB - Ground-dwelling birds in perchlorate-contaminated areas are exposed to perchlorate ion, a known thyroid disruptor, and might be vulnerable to the developmental effects of perchlorate-induced hypothyroidism. We hypothesized that perchlorate-induced hypothyroidism would alter the expression of thyroid responsive genes involved in thyroid hormone (TH) regulation and in the development of target organ function. Japanese quail chicks were exposed to 2000 mg/L ammonium perchlorate in drinking water for 7.5 weeks beginning on day 5 posthatch. Hypothyroidism was evident after 2 weeks of exposure as lower plasma THs and lower TH content in exposed chicks than in controls. The degree of hypothyroidism was increased at 7.5 weeks, as indicated by significant thyroid gland hypertrophy and sustained changes in thyroid function. After 2 weeks of exposure, hypothyroidism increased type 2 5'-deiodinase (D2) mRNA level and decreased Spot 14 (SP14) mRNA level in the liver, whereas D2 mRNA and RC3 mRNA levels in brain were not affected. After 7.5 weeks of exposure, mRNA levels in the exposed group did not differ from those in controls in either the liver or brain, suggesting the responsiveness of these genes to THs decreased during development. These results suggest that the brain, but not the liver, was protected from the effects of hypothyroidism, probably by changes in D2 activity at the protein level and/or regulation of TH entry and exit from the brain. We concluded that perchlorate exposure caused hypothyroidism in young Japanese quail and affected the expression of thyroid-responsive genes during early posthatch development. PMID- 19308638 TI - Modeling protected species habitat and assigning risk to inform regulatory decisions. AB - In the United States, environmental regulatory agencies are required to use "best available" scientific information when making decisions on a variety of issues. However, agencies are often hindered by coarse or incomplete data, particularly as it pertains to threatened and endangered species protection. Stakeholders often agree that more resolute and integrated processes for decision-making are desirable. We demonstrate a process that uses species occurrence data for a federally endangered insect (Karner blue butterfly), a readily available habitat modeling tool, and spatially explicit information about an important Michigan commodity (tart cherries). This case study has characteristics of many protected species regulatory decisions in that species occurrence data were sparse and unequally distributed; regulatory decisions (on pesticide use) were required with potentially significant impacts on a viable agricultural industry; and stakeholder relations were diverse, misinformed, and, in some situations, unjustly contentious. Results from our process include a large-scale, empirically derived habitat suitability map for the focal species and a risk ranking of tart cherry orchards with risk based on the likelihood that pesticide applications will influence the focal protected species. Although the majority (77%) of pesticide-influence zones overlapped Karner blue butterfly habitat, risk scores associated with each orchard were low. Through our process we demonstrated that spatially explicit models can help stakeholders visualize and quantify potential protected species effects. In addition, model outputs can serve to guide field activities (e.g., species surveys and implementation of pesticide buffer zones) that help minimize future effects. PMID- 19308639 TI - Developing rapid methods for analyzing upland riparian functions and values. AB - Regulators protecting riparian areas need to understand the integrity, health, beneficial uses, functions, and values of this resource. Up to now most methods providing information about riparian areas are based on analyzing condition or integrity. These methods, however, provide little information about functions and values. Different methods are needed that specifically address this aspect of riparian areas. In addition to information on functions and values, regulators have very specific needs that include: an analysis at the site scale, low cost, usability, and inclusion of policy interpretations. To meet these needs a rapid method has been developed that uses a multi-criteria decision matrix to categorize riparian areas in Washington State, USA. Indicators are used to identify the potential of the site to provide a function, the potential of the landscape to support the function, and the value the function provides to society. To meet legal needs fixed boundaries for assessment units are established based on geomorphology, the distance from "Ordinary High Water Mark" and different categories of land uses. Assessment units are first classified based on ecoregions, geomorphic characteristics, and land uses. This simplifies the data that need to be collected at a site, but it requires developing and calibrating a separate model for each "class." The approach to developing methods is adaptable to other locations as its basic structure is not dependent on local conditions. PMID- 19308640 TI - Intraoperative blood loss is a critical risk factor for peritoneal recurrence after curative resection of advanced gastric cancer. AB - BACKGROUND: Perioperative blood transfusion has been shown to be associated with poor outcome in various types of malignancy. However, the relationship between the amount of blood loss and specific types of cancer recurrence has not been documented. METHODS: We retrospectively examined the amount of intraoperative blood loss and the recurrence pattern in 146 patients who underwent curative gastrectomy for advanced gastric cancer and assessed the possible correlation between intraoperative blood loss and peritoneal, locoregional, and hematogenous recurrences. RESULTS: The amount of intraoperative blood loss in patients who developed peritoneal recurrence was significantly greater than that in patients without peritoneal recurrence, irrespective of blood transfusion. In contrast, the blood loss was not associated with nodal or hematogenous recurrence. Multivariate analysis demonstrated that large blood loss as well as operative curability B and adjuvant chemotherapy were independent risk factors for peritoneal recurrence and a worse outcome in advanced gastric cancer. CONCLUSIONS: Intraoperative blood loss in curative gastrectomy for advanced gastric cancer may have a specific association with the development of peritoneal recurrence. Surgeons must remember that clean and dry surgery may lessen not only 30-day mortality and morbidity but long-term peritoneal recurrence in gastric cancer. PMID- 19308641 TI - Peer-reviewed, evidence-based analysis of vitamin D and primary hyperparathyroidism. AB - BACKGROUND: Research into the actions of vitamin D on the human body has been increasing at an exponential rate, as has understanding of the impact of vitamin D on various aspects of the endocrine system. Primary hyperparathyroidism (PHPT) is a disease of the endocrine system that can result in debility if not diagnosed. New understanding about the effect of vitamin D on parathyroid hormone regulation may provide an opportunity to better understand the coexistence of PHPT and vitamin D deficiency. METHODS: An evidence-based literature review was based on a PubMed search for research involving vitamin D and primary hyperparathyroidism, and evaluating existing research on vitamin D. The PubMed search included English-language articles published between 1977 and 2008, with a focus on research and analysis completed between 2005 and the present. This study examined recent developments in understanding the relationship between vitamin D and PHPT. This review of existing literature examined the impact of vitamin D insufficiency and deficiency, including the impact on diagnosis of vitamin D insufficiency/deficiency, diagnosis and management of PHPT, and the impact on overall health. The level of evidence was determined according to criteria proposed by Sackett et al. and the grade of recommendation according to the criteria proposed by Heinrich et al. RESULTS: Level III and level IV evidence predominates the vitamin D/PHPT based literature, with several notable Heinrich grade A, B, and C studies available. Additional evaluation of studies' review is provided by reference and by section. CONCLUSIONS: Vitamin D metabolism plays an important role in PHPT etiology, diagnosis, and management. Care of both conditions may have an important impact on overall health in humans. PMID- 19308642 TI - Molecular characterization of the Himalayan mink. AB - A rare color variant of the American mink (Neovison vison), discovered on a ranch in Nova Scotia and referred to as the "marbled" variety, carries a distinctive pigment distribution pattern resembling that found in some other species, e.g., the Siamese cat and the Himalayan mouse. We tested the hypothesis that the color pattern in question-light-colored body with dark-colored points (ears, face, tail, and feet)-is due to a mutation in the melanin-producing enzyme tyrosinase (TYR) that results in temperature-sensitive pigment production. Our study shows that marbled mink carry a mutation in exon 4 of the TYR gene (c.1835C > G) which results in an amino acid substitution (p.H420Q). The location of this substitution corresponds to the amino acid position that is also mutated in the TYR protein of the Himalayan mouse. Thus, the marbled variant is more aptly referred to as the Himalayan mink. PMID- 19308643 TI - Two novel mouse genes mapped to chromosome Yp are expressed specifically in spermatids. AB - The male-specific region of the Y chromosome is evolutionarily predisposed to accumulate genes important for spermatogenesis. Recent work in this laboratory identified two novel Y-linked transcripts that were upregulated in the testis in response to deletions on the chromosome arm Yq. This article reports the further characterisation of these two transcripts and their comparison to related X and autosomal genes. Both map to chromosome arm Yp, outside the Sxr ( b ) deletion interval, both are present in at least two copies on the Y, and both are expressed specifically in spermatids. Given the testicular phenotype of mice with deletions on the Y chromosome, both genes are therefore likely to function in spermatid differentiation. AK006152 is a novel mouse-specific gene with a single potential open reading frame, and it is unusual in that there appears to be no X linked relative. H2al2y is a novel histone in the H2A superfamily and has multiple X-linked relatives and a single autosomal relative in mouse. The presence of a single X-linked copy in rat suggests that H2al amplification is mouse-specific, with the alternative explanation being an earlier amplification followed by gene loss. A phylogenetic analysis of H2al genes together with other H2A genes indicates that H2al is most closely related to the mammalian-specific H2A.Bbd family of histones. Interestingly, K (a)/K (s) analysis indicates that the X and Y members of the H2al family may be under positive selection in mouse, while the autosomal copy is under purifying selection and presumably retains the ancestral function. PMID- 19308644 TI - Dynamical instability determines the effect of ongoing noise on neural firing. AB - At low stimulation rates, electrically stimulated auditory nerve fibers typically fire regularly, in lock-step to the applied stimulus. At high stimulation rates, however, these same fibers fire irregularly. Firing irregularity has been attributed to the random opening and closing of voltage-gated sodium channels at the spike generation site. We demonstrate, however, that the nonlinear dynamics of neural excitation and refractoriness embodied in the FitzHugh-Nagumo (FN) model produce realistic firing irregularity at high stimulus rates, even in the complete absence of ongoing physiological noise. Indeed, we show that ongoing noise can actually regularize the response at low discharge rates. The degree of stimulus-dependent irregularity is determined not so much by the level of ongoing physiological noise as by the dynamical instability. Our work suggests that the dynamical instability, quantified by the Lyapunov exponent, controls neural sensitivity to input signals and to physiological noise, as well the amount of mutual desynchronization between similarly stimulated fibers. This instability, quantified by the value of the Lyapunov exponent, may play a critical role in determining modulation sensitivity and dynamic range in cochlear implants. PMID- 19308645 TI - Total hip arthroplasty in rapidly destructive osteoarthritis of the hip: a case series. AB - Rapidly destructive osteoarthritis (RDO) of the hip is a rare condition characterized by rapid joint degeneration and destruction similar to findings of infection, osteonecrosis, or Charcot disease but without a definitive diagnosis. The cause and natural history of RDO are unclear, but total hip arthroplasty has been utilized as a treatment modality due to the severity of the symptoms. We reviewed retrospectively the records of total hip arthroplasties performed between 1990 and 2003 and identified ten hips in eight patients who fit the profile of the diagnosis of RDO. The mean age at time of surgery was 70. Nine hips were treated with total hip arthroplasty with a hybrid configuration; one hip was treated with a non-cemented total hip arthroplasty. Average follow-up was 6 years with no radiographic evidence of acetabular loosening or osteolysis and no evidence of asymmetric cup wear. One femoral component had evidence of loosening but has not been revised. RDO is an idiopathic condition with no single diagnostic laboratory, pathological, or radiographic finding. A complete preoperative work-up for other causes of hip disease prior to arthroplasty for suspected RDO is necessary to rule out treatable disease. Our series of patients with RDO responded well to hybrid and non-cemented total hip arthroplasty with good clinical and radiographic results. PMID- 19308646 TI - Late perforation and abscess formation at the site of the jejunojejunal anastomosis following laparoscopic Roux-en-Y gastric bypass surgery. AB - Late complications involving the site of the jejunojejunal (J-J) anastomosis are uncommon after laparoscopic Roux-en-Y gastric bypass. We present a case of a perforation at the J-J anastomosis complicated by the formation of an abscess 6 months after surgery. Following clinical and radiological evaluation, the patient underwent an exploratory laparoscopy which had to be converted to an open technique because of technical difficulties. The abscess was drained, the anastomosis was resected, and a new anastomosis was undertaken. When the surgical specimen was inspected, a perforation was found at the site of the previous J-J anastomosis. The patient had an uneventful recovery. To our knowledge, this is the first report of the formation of an abscess due to a perforation at the site of the J-J anastomosis after laparoscopic Roux-en-Y gastric bypass surgery. This complication, although rare, should be taken into account in patients with abdominal pain and systemic inflammatory response syndrome after bariatric surgery. PMID- 19308648 TI - Towards effective Ponseti clubfoot care: the Uganda Sustainable Clubfoot Care Project. AB - Neglected clubfoot is common, disabling, and contributes to poverty in developing nations. The Ponseti clubfoot treatment has high efficacy in correcting the clubfoot deformity in ideal conditions but is demanding on parents and on developing nations' healthcare systems. Its effectiveness and the best method of care delivery remain unknown in this context. The 6-year Uganda Sustainable Clubfoot Care Project (USCCP) aims to build the Ugandan healthcare system's capacity to treat children with the Ponseti method and assess its effectiveness. We describe the Project and its achievements to date (March 2008). The Ugandan Ministry of Health has approved the Ponseti method as the preferred treatment for congenital clubfoot in all its hospitals. USCCP has trained 798 healthcare professionals to identify and treat foot deformities at birth. Ponseti clubfoot care is now available in 21 hospitals; in 2006-2007, 872 children with clubfeet were seen. USCCP-designed teaching modules on clubfoot and the Ponseti method are in use at two medical and three paramedical schools. 1152 students in various health disciplines have benefited. USCCP surveys have (1) determined the incidence of clubfoot in Uganda as 1.2 per 1000 live births, (2) gained knowledge surrounding attitudes, beliefs, and practices about clubfoot across different regions, and (3) identified barriers to adherence to Ponseti treatment protocols. USCCP is now following a cohort of treated children to evaluate its effectiveness in the Ugandan context. LEVEL OF EVIDENCE: Level IV, therapeutic study. See Guidelines for Authors for a complete description of levels of evidence. PMID- 19308649 TI - Synchrotron X-ray fluorescence reveals abnormal metal distributions in brain and spinal cord in spinocerebellar ataxia: a case report. AB - For the first time, synchrotron rapid-scanning X-ray fluorescence (RS-XRF) was used to simultaneously localize and quantify iron, copper, and zinc in spinal cord and brain in a case of spinocerebellar ataxia (SCA). In the normal medulla, a previously undescribed copper enrichment was seen associated with spinocerebellar fibers and amiculum olivae. This region was virtually devoid of all metals in the SCA case. Regions with neuronal loss and gliosis in the cerebellar cortex, inferior olivary, and dentate nuclei and areas showing loss of myelinated fibers were also low in all metals in SCA compared to control. In contrast, the ventral columns of the spinal cord that exhibited only moderate myelin pallor had increased metal levels. Iron and zinc were also elevated in the globus pallidus pars externa in SCA relative to control. We hypothesize that metals increase as part of the initial neurodegenerative process, but once degeneration is advanced, the metal levels drop. This implies a role for multiple metals in SCA neurodegeneration, but further study is required to establish a causative role. We suggest that if these findings are generally true of at least some cases of SCA, not only iron but also copper and zinc should be considered as possible therapeutic targets. PMID- 19308650 TI - Comparison of dobutamine stress echocardiography, dobutamine SPECT, and adenosine SPECT myocardial perfusion imaging in patients with end-stage renal disease. AB - OBJECTIVES: We sought to assess and compare the diagnostic accuracy and prognostic value of dobutamine stress echocardiography (DSE), dobutamine SPECT, and adenosine SPECT myocardial perfusion imaging (MPI) in patients with end-stage renal disease (ESRD). BACKGROUND: The optimal stress imaging modality for patients with ESRD has not yet been determined. METHODS: Forty-nine patients with ESRD underwent DSE, dobutamine SPECT MPI, and adenosine SPECT MPI. The primary endpoint of the trial was concordance between stress tests with respect to the presence or absence of ischemia. RESULTS: Agreement on the presence or absence of ischemia between adenosine SPECT MPI and DSE was 69% (kappa = .25, P = NS). Agreement on the presence or absence of ischemia between adenosine and dobutamine SPECT MPI was 77% (kappa = .37, P = <.009). Summed stress scores for adenosine and dobutamine SPECT MPI studies were highly correlated (r = .9, P = <.0001). DSE and SPECT MPI results provided incremental prognostic information when added to clinical variables. CONCLUSIONS: There is moderate concordance between DSE and adenosine SPECT MPI in ESRD patients referred for stress testing. Interobserver agreement was higher for SPECT MPI compared to DSE. Based on these observations, the optimal approach for diagnosing severe coronary artery disease and assessing risk in patients with ESRD has yet to be determined, but appears to warrant further investigation. PMID- 19308651 TI - Association of the -1082 G/A promoter polymorphism of interleukin-10 gene with the autoantibodies production in patients with rheumatoid arthritis. AB - Interleukin-10 (IL-10) is an immunoregulatory cytokine, usually considered to mediate the downregulation of the inflammatory response in rheumatoid arthritis (RA). Some effects of IL-10 are not anti-inflammatory; for example, the activation of B cells to promote autoantibody production. Allelic polymorphisms located in the promoter region of the IL-10 gene may contribute to the regulation of autoantibodies production. To examine the putative association between the 1082 G/A polymorphism in the promoter region of the IL-10 gene and the susceptibility to disease onset and severity of RA, a total of 144 patients with RA diagnosed according to the revised criteria of the American College of Rheumatology for RA were consecutively recruited into the study. Radiographic progression of RA was scored according to the Sharp/van der Heijde method. Serum levels of rheumatoid factors (RFs) were measured by enzyme-linked immunosorbent assay. Polymerase chain reaction amplification was used for the analysis of the promoter polymorphism of the IL-10 gene. We observed significant differences in genotype distribution of the -1082 G/A polymorphism between IgM RF, IgA RF, and IgG RF positive/negative subgroups of RA patients, with higher prevalence of the GG genotype within IgM RF (Pg = 0.006), IgA RF (Pg = 0.05), and IgG RF (Pg = 0.007) negative RA patients. Results obtained in this study provide the evidence of an association between the -1082 G/A polymorphism in the IL-10 gene promoter and the production of RFs in RA patients. PMID- 19308652 TI - Single port laparoscopic repair of incarcerated ventral hernia. Re: Laparoscopic repair of incarcerated ventral abdominal wall hernias, Shah RH et al. (2008) Hernia 12(5):457-463. PMID- 19308653 TI - Chemical composition and antibacterial activity of essential oils of Eugenia species. AB - The essential oils of the leaves of Eugenia brasiliensis, Eugenia beaurepaireana, and Eugenia umbelliflora were analyzed by GC-MS. The major compounds found in the oil of E. brasiliensis were spathulenol (12.6%) and tau-cadinol (8.7%), of E. beaurepaireana were beta-caryophyllene (8.0%) and bicyclogermacrene (7.2%), and of E. umbelliflora were viridiflorol (17.7%) and beta-pinene (13.2%). These oils were assayed to determine their antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. All of the oils analyzed showed antibacterial activity, ranging from moderate to strong, which was most accentuated for the E. umbelliflora and E. brasiliensis oils, which strongly inhibited the growth of S. aureus giving values of MIC = 119.2 and 156.2 microg/mL, respectively. PMID- 19308654 TI - Inhibitory effects of constituents of Morinda citrifolia seeds on elastase and tyrosinase. AB - A 50% ethanolic extract (MCS-ext) from seeds of Morinda citrifolia ("noni" seeds) showed more potent in vitro inhibition of elastase and tyrosinase, and 1,1 diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity than extracts of M. citrifolia leaves or flesh. Activity-guided fractionation of MCS-ext using in vitro assays led to the isolation of ursolic acid as an active constituent of elastase inhibitory activity. 3,3'-Bisdemethylpinoresinol, americanin A, and quercetin were isolated as active constituents having both tyrosinase inhibitory and radical scavenging activities. Americanin A and quercetin also showed superoxide dismutase (SOD)-like activity. These active compounds were isolated from noni seeds for the first time. PMID- 19308655 TI - Ulcerative and pseudomembranous Aspergillus tracheobronchitis in a patient with acute myeloid leukemia. PMID- 19308656 TI - Low burden of a JAK2-V617F mutated clone in monoclonal haematopoiesis in a Japanese woman with Budd-Chiari syndrome. AB - Approximately one-half of the cases of Budd-Chiari syndrome (BCS) are caused by bcr/abl negative chronic myeloproliferative disorders (CMPDs). Furthermore, a mutation in the Janus kinase protein (JAK2-V617F) is detected in half of the patients with BCS. However, whether the JAK2 mutation is the primary event leading to CMPDs and BCS is controversial. We present a report concerning a young woman who suffered from BCS prior to the onset of CMPDs. Analysis of X-chromosome inactivation patterns in this patient, using the human androgen receptor gene demonstrated monoclonal haematopoiesis in her granulocytes. In contrast, she had a low burden of a JAK2-V617F mutation positive clone among granulocyte populations. These results suggest that the JAK2-V617F mutation occurs after the onset of monoclonal haematopoiesis; thus the V617F mutation of JAK2 may not be the primary event in the induction of BCS. PMID- 19308657 TI - Successful treatment with intravenous colistin for sinusitis, orbital cellulites, and pneumonia caused by multidrug-resistant metallo-beta-lactamase-producing Pseudomonas aeruginosa in a patient with acute myeloid leukemia. AB - The incidence of multidrug-resistant Pseudomonas aeruginosa (MDRPA) and metallo beta-lactamase (MBL)-producing P. aeruginosa has increased worldwide. The treatment options are limited for infectious diseases caused by these two organisms. The use of colistin has been of recent interest in cases involving both types. We report the case of a 74-year-old man with acute myeloid leukemia who was successfully treated with intravenous colistin for maxillary sinusitis and orbital cellulites due to MBL-producing MDRPA during neutropenia, and then for pneumonia caused by the bacteria after the recovery of neutrophil counts. PMID- 19308658 TI - Hereditary spherocytosis and elliptocytosis associated with prosthetic heart valve replacement: rheological study of erythrocyte modifications. AB - The implantation of a prosthetic heart valve (HVP) in patients with hereditary spherocytosis (HS) and hereditary elliptocytosis (HE) is rare, and the changes in the structure and deformability of erythrocytes that follow implantation in these patients have been poorly described. In the present study, the erythrocytes in HS and HE patients with mechanical HVP were compared to the erythrocytes in patients with only congenital membrane defects, in terms of biochemical modifications and rheological behaviour. Integral and cytoskeletal erythrocyte membrane proteins were studied, and blood viscosity (shear rate/shear stress ratio), aggregation ratio [eta(1 s(-1))/eta(200 s(-1))], and red cell visco-elasticity were determined. Valve replacement with a mechanical prosthesis worsened anaemia and resulted in a change in haemolysis, from sub-clinical to evident. The rheological investigation of erythrocytes from HS patients confirmed the characteristic increased viscosity and aggregation ratio and the decreased deformability. The rheological behaviour of erythrocytes from patients with HVP showed a decrease in viscosity and an increase in elastic modulus. In these patients, the prosthesis seems to have induced traumatic damage to the erythrocyte membrane, leading to fragmentation and lysis, which in turn modified rheological parameters. The biochemical and rheological investigation allowed us to understand the clinical and haematological pictures of the patients and to describe the role played by different factors in haemolytic anaemia. PMID- 19308659 TI - Serum sickness with an elevated level of human anti-chimeric antibody following treatment with rituximab in a child with chronic immune thrombocytopenic purpura. AB - Rituximab, a chimeric murine/human monoclonal anti-CD20 antibody, was licensed for the treatment of B-cell lymphoma and has also shown efficacy against autoimmune diseases such as immune thrombocytopenic purpura (ITP). It is relatively safe; however, about 1-20% of patients were reported to have developed rituximab-induced serum sickness, which is more common among patients with autoimmune conditions than among those with hematologic malignancies. Here we describe a pediatric patient with steroid-dependent chronic ITP who presented with arthralgia and fever ten days after the second infusion of rituximab (on day 10), and presented with malaise and maculopapular rash on day 21. Oral prednisolone was started and his symptoms resolved. He had an elevated level of human anti-chimeric antibody (HACA) on day 27; thereafter, the HACA level slowly decreased. To our knowledge, among pediatric patients who received rituximab for chronic ITP, this is the sixth documented case of serum sickness and the only one who manifested an elevated level of HACA. Rituximab is a beneficial treatment option against chronic ITP; however, the risk of serum sickness should be considered. Steroid, usually used for the treatment of serum sickness, may prevent the development of severe serum sickness when administered during and after rituximab treatment. PMID- 19308660 TI - Monosomies 7p and 12p and FLT3 internal tandem duplication: possible markers for diagnosis of T/myeloid biphenotypic acute leukemia and its clonal evolution. AB - Biphenotypic acute leukemia co-expressing T-lymphoid and myeloid markers is rare, accounting for less than 1% of acute leukemias. However, several clinical characteristics including male predominance, frequent lymphadenopathy and unfavorable outcome have been identified. Recurrence of monosomies 7p and/or 12p in T/myeloid biphenotypic acute leukemia has been reported. We treated a patient with T/myeloid biphenotypic acute leukemia showing clonal chromosomal and genetic abnormalities including dic(7;12)(p11;p11) and Fms-like tyrosine kinase 3 (FLT3) internal tandem duplication. Cytogenetic analysis of both bone marrow and lymph node cells disclosed that the patient's lymph node leukemia cells had chromosomal abnormalities in addition to dic(7;12). Our findings suggest that the leukemia cells of systemic lymphadenopathy had evolved as secondary cells from marrow leukemia cells. The patient was successfully treated with induction chemotherapy for acute myeloid leukemia followed by allogeneic bone marrow transplantation. PMID- 19308661 TI - Rare case of femoral artery ramification and origin of the obturator artery. AB - We observed two rare patterns of femoral artery ramification in the bilateral thighs of an 83-year-old male cadaver. In the right thigh, the lateral circumflex femoral and deep circumflex iliac arteries formed a common trunk, while in the left, the medial circumflex femoral, inferior epigastric, and obturator arteries formed a common trunk. In the left pelvis, the obturator artery comprised branches from the inferior epigastric, superior gluteal, and inferior gluteal arteries. We describe this rare case and discuss the genesis and clinical significance of these variations. PMID- 19308662 TI - Osmosensing and osmosignaling in Corynebacterium glutamicum. AB - The Gram-positive soil bacterium Corynebacterium glutamicum is used in microbial biotechnology for the large-scale production of amino acids, e.g., L: -glutamate and L: -lysine. We have studied the response of this organism to hyperosmotic challenge at the level of both transcription and protein activity. Two systems responding to hyperosmotic stress in C. glutamicum are reviewed here, the two component system MtrAB and the glycine-betaine uptake system BetP. The osmosensory two-component system consists of the membrane-bound histidine kinase MtrB and the soluble response regulator MtrA. MtrB was shown to perceive a so far unknown physical stimulus related to hyperosmotic stress via the cytoplasmically oriented phosphorylation domain, and to transduce the signal to the DNA via MtrA. The secondary active transporter BetP takes up betaine in cotransport with two Na(+) ions. BetP responds to hyperosmotic stress by increased transcription mediated via MtrAB signaling, and by instant activation of transport. In the mechanism of BetP activation, the C-terminal, regulatory domain of BetP, the cytoplasmic concentration of K(+), and negative membrane surface charges are involved. The molecular mechanism of the activation process is discussed in relation to the recently published X-ray structure of BetP. PMID- 19308664 TI - Evolution of genitalia: theories, evidence, and new directions. AB - Many hypotheses have been proposed to explain why male intromittent genitalia consistently tend to diverge more rapidly than other body traits of the same individuals in a wide range of animal taxa. Currently the two most popular involve sexual selection: sexually antagonistic coevolution (SAC) and cryptic female choice (CFC). A review of the most extensive attempts to discriminate between these two hypotheses indicates that SAC is not likely to have played a major role in explaining this pattern of genital evolution. Promising lines for future, more direct tests of CFC include experimental modification of male genital form and female sensory abilities, analysis of possible male-female dialogues during copulation, and direct observations of genital behavior. PMID- 19308665 TI - Prediction of breast cancer metastasis by genomic profiling: where do we stand? AB - Current concepts conceive "breast cancer" as a complex disease that comprises several very different types of neoplasms. Nonetheless, breast cancer treatment has considerably improved through early diagnosis, adjuvant chemotherapy, and endocrine treatments. The limited prognostic power of classical classifiers determines considerable over-treatment of women who either do not benefit from, or do not at all need, chemotherapy. Several gene expression based molecular classifiers (signatures) have been developed for a more reliable prognostication. Gene expression profiling identifies profound differences in breast cancers, most probably as a consequence of different cellular origin and different driving mutations and can therefore distinguish the intrinsic propensity to metastasize. Existing signatures have been shown to be useful for treatment decisions, although they have been developed using relatively small sample numbers. Major improvements are expected from the use of large datasets, subtype specific signatures and from the re-introduction of functional information. We show that molecular signatures encounter clear limitations given by the intrinsic probabilistic nature of breast cancer metastasis. Already today, signatures are, however, useful for clinical decisions in specific cases, in particular if the personal inclination of the patient towards different treatment strategies is taken into account. PMID- 19308666 TI - Nuclear magnetic resonance and circular dichroism study of metastin (Kisspeptin 54) structure in solution. AB - KISS1 was first discovered as a metastasis suppressor, but also plays crucial roles in the onset of puberty. The KISS1 gene encodes a secreted protein of 145 amino acids that exhibits no sequence similarity with any known proteins. KISS1 protein is proteolytically processed to generate a number of so-called kisspeptins (KP), the most well characterized is known as KP-54 or metastin. KP 54 is carboxy-terminally amidated and binds to and activates the KISS1 receptor (KISS1R). The current studies were undertaken in order to determine structure of KP-54 using nuclear magnetic resonance and circular dichroism. KP-54 is mostly disordered both in water and in trifluoroethanol/water mixed solvent, with no structural motifs. In sodium dodecyl sulfate micelles, KP-54 remains mostly disordered except for a small increase in helical propensity (from 3.7% in water to 9.9% in micelles). Despite this apparent lack of structure, KP-54 is biologically active. The intrinsic disorder of KP-54 may confer advantages in its ability to recognize and bind a wide range of target proteins. PMID- 19308667 TI - Monitoring of cardiovascular reactivity to cold stress using digital volume pulse characteristics in health and diabetes. AB - OBJECTIVE: Cold pressor test (CPT) assesses sympathetic reactivity by the rise in diastolic blood pressure secondary to the vasoconstriction during immersion of hand in cold water. Presently, monitoring of vascular reactivity in health and diabetes during CPT has been attempted by objective measures of Photoplethysmogram (PPG) that include amplitude, upstroke-slope, pulse timings and pulse transit time (PTT). METHODS: Finger-PPG characteristics were studied before and during CPT (1 min) in 11 healthy volunteers and 10 diagnosed Type 2 Diabetes Mellitus (DM) patients. In controls, the recordings were continued for 5 min after CPT. RESULTS: The amplitude of PPG significantly decreased due to cold stress in both control and DM groups (P < 0.0001 and P < 0.003, respectively). However, the decrease in amplitude was significantly lesser (0.42 +/- 0.08 nu vs. 0.25 +/- 0.03 nu, P = 0.04) in DM group than controls. The slope response of PPG resembled the amplitude. PTT was significantly shortened in control and DM groups (180.0 +/- 3.8 ms vs. 187.1 +/- 3.9 ms, P < 0.006, 177.7 +/- 7.0 ms vs. 192.9 +/- 5.6 ms, P = 0.002, respectively) during CPT as compared to baseline. However, the decrease in PTT was significantly higher (-15.2 +/- 3.4 ms vs. -6.0 +/- 1.9 ms, P = 0.03) in DM patients than controls. No significant differences were noticed in Delta changes of peak-to-peak interval, crest time and decay time of PPG between the two groups. CONCLUSION: This preliminary study suggests that the collective responses of PPG amplitude and PTT can be used to objectively quantify the sympathetic reactivity to cold stress in health as well as to detect the deficits of vascular reactivity in diabetes. Further studies would substantiate the simple PPG technique in quantifying the neuronal and vascular dysfunction. PMID- 19308668 TI - Professional ethics in pharmacy practice: developing a psychometric measure of moral reasoning. AB - OBJECTIVE: To develop and validate a psychometric measure of cognitive moral development in professional ethics in pharmacy. SETTING: Pharmacy practice in Australia. METHOD: A psychometric instrument, the Professional Ethics in Pharmacy (PEP) test, was developed and validated following a systematic procedure. The theoretical foundation of the instrument was based on a hypothesised theory of cognitive moral development in professional ethics, which was integrated into a selection of scenarios experienced in practice by pharmacists in NSW, Australia. The PEP, along with the well established DIT test, was mailed in the form of a questionnaire to a randomly selected sample of 1,500 practising pharmacists. Data collected from returned questionnaires were statistically analysed to establish validity and reliability of the instrument. MAIN OUTCOME MEASURES: The P-score calculated for each participant from DIT and PEP data and defined as a measure of principled moral reasoning, represented the main outcome measure for statistical testing. Pearson's correlation coefficient was used to determine the strength of the relationship of the PEP with the DIT (regarded as the "gold standard") in order to establish criterion and concurrent validity. Factor analysis was used to investigate construct validity. Cronbach's Alpha, a measure of reliability of the instrument, was used for indicating internal consistency. Linear regression models further investigated construct validity in relation to predictors of moral reasoning. RESULTS: Face and content validity were established by pilot and peer review. Pearson's coefficient of 0.53 indicated an acceptable level of concurrent validity. Factor analysis yielded factors closely related to the theoretical stages of cognitive moral development hypothesised, which indicated construct validity. Cronbach's alpha of 0.75 demonstrated the reliability of the instrument, and linear regression models provided further evidence of construct validity. The PEP was established as a robust instrument on several dimensions of validity and reliability. CONCLUSION: The validated PEP test has the potential to provide the pharmacy profession with valuable information for use education and research. The validation process also provided evidence supporting the hypothesis that moral reasoning in professional ethics in pharmacy is a developmental process, which has profound implications for furthering the understanding of professional behaviour. PMID- 19308669 TI - Parity-violating energy shifts of Murchison L-amino acids are consistent with an electroweak origin of meteorite L-enantiomeric excesses. AB - In 1996, four alpha-methyl amino acids in the Murchison meteorite--L-isovaline, L alpha-methylnorvaline, L-alpha-methyl-allo-isoleucine and L-alpha-methyl isoleucine--were found to show significant enantiomeric excesses of the L form, ranging from 2% to 9%. Their deuterium to hydrogen isotope ratios suggest they formed in the pre-solar interstellar gas cloud rather than during a later aqueous processing phase on the asteroid parent body. In this paper we apply the techniques of the preceding two papers to compute the parity-violating energy shifts of these amino acids. We find that, in the gas phase, the PVESs of the neutral L forms of all four Murchison alpha-methyl amino acids are decisively negative, and there is even some correlation between the magnitudes of the L excesses and the magnitudes of the PVESs--all of which is at least consistent with an electroweak origin of the Murchison enantiomeric excesses. PMID- 19308670 TI - Postmortem studies on a patient with mucopolysaccharidosis type I: histopathological findings after one year of enzyme replacement therapy. AB - Deficiency of lysosomal alpha-L-iduronidase results in systemic accumulation of glycosaminoglycans (GAGs). Cardiac lesions due to accumulation of GAGs include hypertrophic cardiomyopathy, valvular insufficiency/stenosis, and coronary artery stenosis due to intimal proliferation. Cardiac dysfunction is one of the most common causes of death in patients with mucopolysaccharidosis type I (MPS I). Enzyme replacement therapy (ERT) with laronidase has shown clear effects in reduction of hepatomegaly and it has been unclear whether ERT could improve or prevent the cardiac lesions. Postmortem findings in a 3 1/2-year-old boy diagnosed with MPS I at age 2 years are described. He received ERT with laronidase at 100 U/kg/week for one year. He suddenly developed cardiorespiratory failure and died the next day after C2-3 spinal surgery for instability. Postmortem examination showed hypertrophic cardiomyopathy, severe aortic valve and mitral valve thickening with shortened chordae, and endocardial fibroelastosis. Histology of the cardiac tissue revealed increased perivascular and interstitial connective tissue in the myocardium and intimal thickening causing stenosis in the cardiac vessels. Electron-microscopic (EM) studies of the thickened endocardium revealed numerous histiocytes with enlarged lysosomes. EM examination of the liver and the cardiac muscle revealed no accumulation of GAGs. ERT with laronidase showed clear effects in removing GAGs from the liver and the cardiac muscle. However, it did not show a clear effect on the thickened endocardium, myocardial perivascular and interstitial connective tissue or intimal thickening in the epicardial vessels. PMID- 19308671 TI - Identification of Penicillium marneffei in Paraffin-Embedded Tissue Using Nested PCR. AB - Penicillium marneffei is one of the unique thermally dimorphic fungi in Penicillium species that causes a disseminated, progressive and life threatening infection in immunocompromised patients. The diagnosis of Penicilliosis marneffei depends on culture that may delay the treatment due to the time-consuming process. In the present study, we evaluated the specificity and sensitivity of nested PCR to identify Penicillium marneffei from paraffin-embedded tissue. Two sets of oligonucleotide primers were derived from the sequence of 18S rRNA of Penicillium marneffei. The outer primers (RRF1 and RRH1) were specific to fungi. The inner primers (Pm1 and Pm2) were specific to Penicillium marneffei. The specific fragment of approximately 400 bp was amplified from all paraffin embedded tissues from 14 patients with Penicilliosis marneffei and 10 bamboo rats. The detectable DNA concentration of single PCR and nested PCR were 14 pg/microl and 14 fg/microl, respectively. Further studies are required in order to use nested PCR for early diagnosis of the disease. PMID- 19308672 TI - Antifungal therapy for invasive fungal diseases in allogeneic stem cell transplant recipients: an update. AB - Invasive fungal diseases (IFDs) remain a major cause of morbidity and mortality in allogeneic stem cell transplant (SCT) recipients. While the most common pathogens are Candida spp. and Aspergillus spp., the incidence of infections caused by non-albicans Candida species as well as molds such as Zygomycetes has increased. For many years, amphotericin B deoxycholate (AMB-D) was the only available antifungal for the treatment of IFDs. Within the past decade, there has been a surge of new antifungal agents developed and added to the therapeutic armamentarium. Lipid-based formulations of amphotericin B provide an effective and less nephrotoxic alternative to AMB-D. Voriconazole has now replaced AMB-D as first choice for primary therapy of invasive aspergillosis (IA). Another extended spectrum triazole, posaconazole, also appears to be a promising agent in the management of zygomycosis, refractory aspergillosis, and for prophylaxis. Members of the newest antifungal class, the echinocandins, are attractive agents in select infections due to their safety profile, and are a more attractive option compared to AMB-D as initial treatment for invasive candidiasis and (based on one study) challenge fluconazole for superiority in management with this mycoses. However, challenges do exist among these newer agents in very high-risk individuals like allogeneic SCT recipients, which may include adverse drug events, drug-drug interactions, variability in oral absorption, and availability of alternative formulations. The addition of newer agents has also stimulated interest in the potential application of combination therapy in serious, life threatening infections. However, adequate studies are not available for most IFDs; thus, the clinical use of combination therapy is not evidenced based on most cases and preciseness in its use is uncertain. Finally, therapeutic drug monitoring of select antifungals (notably posaconazole and voriconazole) may play an increasing role due to significant interpatient variability in serum concentrations after standard doses. PMID- 19308673 TI - Molecular cloning and expression of a bush related CmV1 gene in tropical pumpkin. AB - A bush-type plant was selected from tropical pumpkin 'cga' (Cucurbita moschata Duchesne) in order to study the vine development in C. moschata. In this study, a novel gene encoding NADH dehydrogenase was isolated from the vine line (cgaV) of C. moschata, that was not expressed in the near isogenic bush line (cgaBu). This gene, designated as CmV1 (C. moschata vine 1), was 545 bp in length and was composed of a 477 bp open reading frame, which had 99% nucleotide similarity to the chloroplast ndhJ gene for NADH dehydrogenase subunit J from Brassica oleracea. The deduced amino acid sequence of CmV1 had 99% similarity to NADH dehydrogenase subunit J from Arabidopsis and had 98% similarity to NADH dehydrogenase subunit from Barbarea verna. Analysis of the basic characteristics of the CmV1 protein revealed that it has one Respiratory chain NADH dehydrogenase 30 kD subunit signature, three N-myristoylation sites, one Casein kinase II phosphorylation site, and one Protein kinase C phosphorylation site. Reverse transcriptase-PCR analysis showed that CmV1 was expressed at a high level in the internodes and hypocotyls and was expressed stronger in elongating internodes than in fully expanded internodes. In conclusion, results obtained in the present study suggest that CmV1 gene might play important roles in vine elongation of tropical pumpkin. PMID- 19308674 TI - Vascular smooth muscle cell expression of ectonucleotidase CD39 (ENTPD1) is required for neointimal formation in mice. AB - Vascular smooth muscle cell (VSMC) migration and proliferation are critical steps in the pathogenesis of atherosclerosis, post-angioplasty restenosis, neointimal hyperplasia, and chronic allograft rejection. Extracellular nucleotides are known to influence both migration and proliferation of VSMC. Although it is well established that vascular endothelial Cd39/ENTPD1 regulates blood nucleotide concentrations, whether Cd39 associated with VSMC also impacts vascular wall pathology has not been investigated. The objective of this paper is to determine levels of expression of Cd39 on VSMC and functional consequences of gene deletion in vitro and in vivo. Cd39 is the major ectonucleotidase in VSMC, as shown by substantive decreases in ecto-ATPase and -ADPase activity in Cd39-null cells compared to wild type. Significant decreases in neointimal lesion formation are observed in Cd39-null mice at 21 days post arterial balloon injury. Stimulated Cd39-null VSMC have pronounced proliferative responses in vitro. However, using Transwell systems, we show that Cd39-null VSMC fail to migrate in response to ATP, UTP, and PDGF. Cd39 is the dominant ectonucleotidase expressed by VSMC. Deletion of Cd39 in mice results in decreased neointimal formation after vascular injury and is associated with impaired VSMC migration responses in vitro. PMID- 19308675 TI - Effects of extracellular nucleotides on renal tubular solute transport. AB - A range of P2 receptor subtypes has been identified along the renal tubule, in both apical and basolateral membranes. Furthermore, it has been shown that nucleotides are released from renal tubular cells, and that ectonucleotidases are present in several nephron segments. These findings suggest an autocrine/paracrine role for nucleotides in regulating tubular function. The present review catalogues the known actions of extracellular nucleotides on tubular solute transport. In the proximal tubule, there is firm evidence that stimulation of apical P2Y(1) receptors inhibits bicarbonate reabsorption, whilst basolaterally applied ATP has the opposite effect. Clearance studies suggest that systemic diadenosine polyphosphates profoundly reduce proximal tubular fluid transport, through as yet unidentified P2 receptors. To date, only circumstantial evidence is available for an action of nucleotides on transport in the loop of Henle; and no studies have been made on native distal tubules, though observations in cell lines suggest an inhibitory effect on sodium, calcium and magnesium transport. The nephron segment most studied is the collecting duct. Apically applied nucleotides inhibit the activity of small-conductance K(+) channels in mouse collecting duct, apparently through stimulation of P2Y(2) receptors. There is also evidence, from cell lines and native tissue, that apically (and in some cases basolaterally) applied nucleotides inhibit sodium reabsorption. In mice pharmacological profiling implicates P2Y(2) receptors; but in rats, the receptor subtype(s) responsible is/are unclear. Recent patch-clamp studies in rat collecting ducts implicate apical P2Y and P2X subtypes, with evidence for both inhibitory and stimulatory effects. Despite considerable progress, clarification of the physiological role of the tubular P2 receptor system remains some way off. PMID- 19308676 TI - Nuclear localization of CXCR4 determines prognosis for colorectal cancer patients. AB - Chemokines and their receptors are implicated in formation of colorectal cancer metastases. Especially CXCR4 is an important factor, determining migration, invasiveness, metastasis and proliferation of colorectal cancer cells. Object of this study was to determine expression of CXCR4 in tumor tissue of colorectal cancer patients and associate CXCR4 expression levels to clinicopathological parameters. Levels of CXCR4 expression of a random cohort of patients, who underwent primary curative resection of a colorectal carcinoma, were retrospectively determined by quantitative real-time RT-PCR and semi-quantitative analyses of immunohistochemical stained paraffin sections. Expression levels were associated to clinicopathological parameters. Using RT-PCR we found that a high expression of CXCR4 in the primary tumor was an independent prognostic factor for a poor disease free survival (p = 0.03, HR: 2.0, CI = 1.1-3.7). Immunohistochemical staining showed that nuclear distribution of CXCR4 in the tumor cells was inversely associated with disease free and overall survival (p = 0.04, HR: 2.6, CI = 1.0-6.2), while expression in the cytoplasm was not associated with prognosis. In conclusion, our study showed that a high expression of nuclear localized CXCR4 in tumor cells is an independent predictor for poor survival for colorectal cancer patients. PMID- 19308677 TI - Dual FGF-2 and intergrin alpha5beta1 signaling mediate GRAF-induced RhoA inactivation in a model of breast cancer dormancy. AB - Interactions with the bone marrow stroma regulate dormancy and survival of breast cancer micrometastases. In an in vitro model of dormancy in the bone marrow, we previously demonstrated that estrogen-dependent breast cancer cells are partially re-differentiated by FGF-2, re-express integrin alpha5beta1 lost with malignant transformation and acquire an activated PI3K/Akt pathway. Ligation of integrin alpha5beta1 by fibronectin and activation of the PI3K pathway both contribute to survival of these dormant cells. Here, we investigated mechanisms responsible for the dormant phenotype. Experiments demonstrate that integrin alpha5beta1 controls de novo cytoskeletal rearrangements, cell spreading, focal adhesion kinase rearrangement to the cell perimeter and recruitment of a RhoA GAP known as GRAF. This results in the inactivation of RhoA, an effect which is necessary for the stabilization of cortical actin. Experiments also demonstrate that activation of the PI3K pathway by FGF-2 is independent of integrin alpha5beta1 and is also required for cortical actin reorganization, GRAF membrane relocalization and RhoA inactivation. These data suggest that GRAF-mediated RhoA inactivation and consequent phenotypic changes of dormancy depend on dual signaling by FGF-2 initiated PI3K activation and through ligation of integrin alpha5beta1 by fibronectin. PMID- 19308678 TI - Genotype-dependent tumor regression in Marek's disease mediated at the level of tumor immunity. AB - Marek's disease (MD) of chickens is a unique natural model of Hodgkin's and Non Hodgkin's lymphomas in which the neoplastically-transformed cells over-express CD30 (CD30(hi)) antigen. All chicken genotypes can be infected with MD virus and develop microscopic lymphomas. From 21 days post infection (dpi) microscopic lymphomas regress in resistant chickens but, in contrast, they progress to gross lymphomas in susceptible chickens. Here we test our hypothesis that in resistant chickens at 21 dpi the tissue microenvironment is pro T-helper (Th)-1 and compatible with cytotoxic T lymphocyte (CTL) immunity but in susceptible lines it is pro Th-2 or pro T-regulatory (T-reg) and antagonistic to CTL immunity. We used the B2, non-MHC-associated, MD resistance/susceptibility system (line [L]6(1)/line [L]7(2)) and quantified the levels of key mRNAs that can be used to define Th-1 (IL-2, IL-12, IL-18, IFNgamma), Th-2 (IL-4, IL-10) and T-reg (TGFbeta, GPR-83, CTLA-4, SMAD-7) lymphocyte phenotypes. We measured gene expression in both whole tissues (represents tissue microenvironment and tumor microenvironment) and in the lymphoma lesions (tumor microenvironment) themselves. Gene ontology-based modeling of our results shows that the dominant phenotype in whole tissue as well as in microscopic lymphoma lesions, is pro T reg in both L6(1) and L7(2) but a minor pro Th-1 and anti Th-2 tissue microenvironment exists in L6(1) whereas there is an anti Th-1 and pro Th-2 tissue microenvironment in L7(2). The tumor microenvironment per se is pro T-reg, anti Th-1 and pro Th-2 in both L6(1) and L7(2). Together our data suggests that the neoplastic transformation is essentially the same in both L6(1) and L7(2) and that resistance/susceptibility is mediated at the level of tumor immunity in the tissues. PMID- 19308679 TI - Identification of molecular distinctions between normal breast-associated fibroblasts and breast cancer-associated fibroblasts. AB - Stromal fibroblasts influence the behavior of breast epithelial cells. Fibroblasts derived from normal breast (NAF) inhibit epithelial growth, whereas fibroblasts from breast carcinomas (CAF) have less growth inhibitory capacity and can promote epithelial growth. We sought to identify molecules that are differentially expressed in NAF versus CAF and potentially responsible for their different growth regulatory abilities. To determine the contribution of soluble molecules to fibroblast-epithelial interactions, NAF were grown in 3D, transwell or direct contact co-cultures with MCF10AT epithelial cells. NAF suppressed proliferation of MCF10AT in both direct contact and transwell co-cultures, but this suppression was significantly greater in direct co-cultures, indicating involvement of both soluble and contact factors. Gene expression profiling of early passage fibroblast cultures identified 420 genes that were differentially expressed in NAF versus CAF. Of the eight genes selected for validation by real time PCR, FIBULIN 1, was overexpressed in NAF, and DICKKOPF 1, NEUREGULIN 1, PLASMINOGEN ACTIVATOR INHIBITOR 2, and TISSUE PLASMINOGEN ACTIVATOR were overexpressed in CAF. A higher expression of FIBULIN 1 in normal- than cancer associated fibroblastic stroma was confirmed by immunohistochemistry of breast tissues. Among breast cancers, stromal expression of Fibulin 1 protein was higher in estrogen receptor alpha-positive cancers and low stromal expression of Fibulin 1 correlated with a higher proliferation of cancer epithelial cells. In conclusion, expression profiling of NAF and CAF cultures identified many genes with potential relevance to fibroblast-epithelial interactions in breast cancer. Furthermore, these early passage fibroblast cultures can be representative of gene expression in stromal fibroblasts in vivo. PMID- 19308680 TI - Metastasis suppressors and the tumor microenvironment. AB - The most dangerous attribute of cancer cells is their ability to metastasize. Throughout the process of metastasis, tumor cells interact with other tumor cells, host cells and extracellular molecules. This brief review explores how a new class of molecules - metastasis suppressors - regulate tumor cell microenvironmental interactions. Data are presented which demonstrate that metastasis suppressors act at multiple steps of the metastatic cascade. A brief discussion for how metastasis suppressor regulation of cellular interactions might be exploited is presented. PMID- 19308681 TI - The epigenetic influence of tumor and embryonic microenvironments: how different are they? AB - The microenvironment is being increasingly recognized as a critical component in tumor progression and metastases. As such, the bi-directional signaling of extracellular mediators that promote tumor growth within the microenvironment is a focus of intense scrutiny. Interestingly, there are striking similarities between the phenotypes of aggressive tumor and embryonic stem cells, particularly with respect to specific signaling pathways underlying their intriguing plasticity. Here, we demonstrate the epigenetic influence of the hESC microenvironment on the reprogramming of aggressive melanoma cells using an innovative 3-D model. Specifically, our laboratory has previously demonstrated the redifferentiation of these melanoma cells to a more melanocyte-like phenotype (Postovit et al., Stem Cells 24(3):501-505, 2006), and now we show the loss of VE Cadherin expression (indicative of a plastic vasculogenic phenotype) and the loss of Nodal expression (a plasticity stem cell marker) in tumor cells exposed to the hESC microenvironment. Further studies with the 3-D culture model revealed the epigenetic influence of aggressive melanoma cells on hESCs resulting in the down regulation of plasticity markers and the emergence of phenotype-specific genes. Additional studies with the aggressive melanoma conditioned matrix microenvironment demonstrated the transdifferentiation of normal melanocytes into melanoma-like cells exhibiting a vasculogenic phenotype. Collectively, these studies have advanced our understanding of the epigenetic influence associated with the microenvironments of hESCs and aggressive melanoma cells, and shed new light on their therapeutic implications. Moreover, we have a better appreciation of the convergence of embryonic and tumorigenic signaling pathways that might stimulate further consideration of targeting Nodal in aggressive tumor cells resulting in a down-regulation of tumorigenic potential and plasticity. PMID- 19308682 TI - Bone marrow microenvironment and tumor progression. AB - The bone marrow constitutes an unique microenvironment for cancer cells in three specific aspects. First, the bone marrow actively recruits circulating tumor cells where they find a sanctuary rich in growth factors and cytokines that promote their proliferation and survival. When in the bone marrow, tumor cells profoundly affect the homeostasis of the bone and the balance between osteogenesis and osteolysis. As a consequence, growth and survival factors normally sequestered into the bone matrix are released, further fueling cancer progression. Second, tumor cells actively recruit bone marrow-derived precursor cells into their own microenvironment. When in the tumors, these bone marrow derived cells contribute to an inflammatory reaction and to the formation of the tumor vasculature. Third, bone marrow-derived cells can home in distant organs, where they form niches that attract circulating tumor cells. Our understanding of the contribution of the bone marrow microenvironment to cancer progression has therefore dramatically improved over the last few years. The importance of this new knowledge cannot be underestimated considering that the vast majority of cancer treatments such as cytotoxic and myeloablative chemotherapy, bone marrow transplantation and radiation therapy inflict a trauma to the bone marrow microenvironment. How such trauma affects the influence that the bone marrow microenvironment exerts on cancer is still poorly understood. In this article, the reciprocal relationship between the bone marrow microenvironment and tumor cells is reviewed, and its potential impact on cancer therapy is discussed. PMID- 19308683 TI - Significance of toll-like receptors expression in tumor growth and spreading: a short review. AB - Toll-like receptors (TLRs) are considered now as crucial sensors of innate immunity. Their role in the recognition of pathogens and the initiation of adaptive immune responses against them is well known. However, in last years TLRs have been identified on several tumor cells, including human malignancies. Their expression in cancer was found to be twofold: either promoting or inhibiting tumor progression. It was also demonstrated that several TLRs agonists, either natural or synthetic ones, may have beneficial effect on tumor-mediated disease, leading to potentiation of immune response to tumor-associated antigens. TLR agonist linked tumor immunotherapy is still in nascent state, but growing rapidly, also in the area of common human malignancies. To date, the most promising and the most frequently studied interaction in tumor immunotherapy trials seems to be TLR9 and its synthetic agonists. PMID- 19308684 TI - Regulation of tumor progression by extracellular galectin-3. AB - The relationship between a tumor cell and its microenvironment is bi-directional. The proteins expressed by the tumor cells alter the signatures on the seemingly normal stromal cells within the microenvironment, while the tumor cell signatures reflect the changes that occur as these cells interact with the host microenvironment. Galectin-3 is a carbohydrate-binding protein that is over expressed in a variety of tumors and immune cells in response to various stimuli. Ever since its discovery, it has been associated with cell and extracellular matrix interactions. However, in the last decade, an extensive accumulation of data has changed the perspective of this multifunctional protein. The unique structure of this protein, consisting of a carbohydrate-binding domain and a matrix metalloproteinase cleavable domain, enables it to interact with a plethora of ligands in a carbohydrate-dependent or independent manner. It is now becoming evident that galectin-3 is involved with a variety of extracellular functions like cell adhesion, migration, invasion, angiogenesis, immune functions, apoptosis and endocytosis. Galectin-3 is a substrate for matrix metalloproteinases and its cleavage plays an important role in tumor progression and can be used as a surrogate diagnostic marker for in vivo MMP activity. PMID- 19308685 TI - A dialogue between the hypoxia-inducible factor and the tumor microenvironment. AB - The hypoxia-inducible factor is the key protein responsible for the cellular adaptation to low oxygen tension. This transcription factor becomes activated as a result of a drop in the partial pressure of oxygen, to hypoxic levels below 5% oxygen, and targets a panel of genes involved in maintenance of oxygen homeostasis. Hypoxia is a common characteristic of the microenvironment of solid tumors and, through activation of the hypoxia-inducible factor, is at the center of the growth dynamics of tumor cells. Not only does the microenvironment impact on the hypoxia-inducible factor but this factor impacts on microenvironmental features, such as pH, nutrient availability, metabolism and the extracellular matrix. In this review we discuss the influence the tumor environment has on the hypoxia-inducible factor and outline the role of this factor as a modulator of the microenvironment and as a powerful actor in tumor remodeling. From a fundamental research point of view the hypoxia-inducible factor is at the center of a signaling pathway that must be deciphered to fully understand the dynamics of the tumor microenvironment. From a translational and pharmacological research point of view the hypoxia-inducible factor and its induced downstream gene products may provide information on patient prognosis and offer promising targets that open perspectives for novel "anti-microenvironment" directed therapies. PMID- 19308686 TI - Role of cancer microenvironment in metastasis: focus on colon cancer. AB - One person on three will receive a diagnostic of cancer during his life. About one third of them will die of the disease. In most cases, death will result from the formation of distal secondary sites called metastases. Several events that lead to cancer are under genetic control. In particular, cancer initiation is tightly associated with specific mutations that affect proto-oncogenes and tumour suppressor genes. These mutations lead to unrestrained growth of the primary neoplasm and a propensity to detach and to progress through the subsequent steps of metastatic dissemination. This process depends tightly on the surrounding microenvironment. In fact, several studies support the point that tumour development relies on a continuous cross-talk between cancer cells and their cellular and extracellular microenvironments. This signaling cross-talk is mediated by transmembrane receptors expressed on cancer cells and stromal cells. The aim of this manuscript is to review how the cancer microenvironment influences the journey of a metastatic cell taking liver invasion by colorectal cancer cells as a model. PMID- 19308687 TI - Abnormal growth factor/cytokine network in gastric cancer. AB - Gastric cancer cells express a broad spectrum of the growth factor/cytokine receptor systems that organize the complex interaction between cancer cells and stromal cells in tumor microenvironment, which confers cell growth, apoptosis, morphogenesis, angiogenesis, progression and metastasis. However, these abnormal growth factor/cytokine networks differ in the two histological types of gastric cancer. Importantly, activation of nuclear factor-kB pathway by Helicobacter pylori infection may act as a key player for induction of growth factor/cytokine networks in gastritis and pathogenesis of gastric cancer. Better understanding of these events will no doubt provide new approaches for biomarkers of diagnosis and effective therapeutic targeting of gastric cancer. PMID- 19308688 TI - Matricellular proteins produced by melanocytes and melanomas: in search for functions. AB - Matricellular proteins are modulators of cell-matrix interactions and cellular functions. The group includes thrombospondin, osteopontin, osteonectin/SPARC, tenascin, disintegrins, galectins and CCN proteins. The production of matricellular proteins such as osteopontin, SPARC or tenascin is highly upregulated in melanoma and other tumors but little is known about their functions in tumor growth, survival, and metastasis. The distribution pattern of CCN3 differs from most other matricellular proteins, such that it is produced abundantly by normal melanocytes, but is not significantly expressed in melanoma cells. CCN3 is known to inhibit melanocyte proliferation and stimulate adhesion to collagen type IV, the main component of the basement membrane. CCN3 has a unique role in securing adhesion of melanocytes to the basement membrane distinct from other melanoma-produced matricellular proteins which act as de-adhesive molecules and antagonists of focal adhesion. Qualitative and quantitative changes in matricellular protein expression contribute to melanoma progression similar to the E-cadherin to N-cadherin class switch, allowing melanoma cells to escape from keratinocyte control. PMID- 19308689 TI - Emerging roles of PAR-1 and PAFR in melanoma metastasis. AB - Melanoma growth, angiogenesis and metastatic progression are strongly promoted by the inflammatory tumor microenvironment due to high levels of cytokine and chemokine secretion by the recruited inflammatory and stromal cells. In addition, platelets and molecular components of procoagulant pathways have been recently emerging as critical players of tumor growth and metastasis. In particular, thrombin, through the activity of its receptor protease-activated receptor-1 (PAR 1), regulates tumor cell adhesion to platelets and endothelial cells, stimulates tumor angiogenesis, and promotes tumor growth and metastasis. Notably, in many tumor types including melanoma, PAR-1 expression directly correlates with their metastatic phenotype and is directly responsible for the expression of interleukin-8, matrix metalloproteinase-2 (MMP-2), vascular endothelial growth factor, platelet-derived growth factor, and integrins. Another proinflammatory receptor-ligand pair, platelet-activating factor (PAF) and its receptor (PAFR), have been shown to act as important modulators of tumor cell adhesion to endothelial cells, angiogenesis, tumor growth and metastasis. PAF is a bioactive lipid produced by a variety of cells from membrane glycerophospholipids in the same reaction that releases arachidonic acid, and can be secreted by platelets, inflammatory cells, keratinocytes and endothelial cells. We have demonstrated that in metastatic melanoma cells, PAF stimulates the phosphorylation of cyclic adenosine monophosphate response element-binding protein (CREB) and activating transcription factor 1 (ATF-1), which results in overexpression of MMP-2 and membrane type 1-MMP (membrane type 1-MMP). Since only metastatic melanoma cells overexpress CREB/ATF-1, we propose that metastatic melanoma cells are better equipped than their non-metastatic counterparts to respond to PAF within the tumor microenvironment. The evidence supporting the hypothesis that the two G protein coupled receptors, PAR-1 and PAFR, contribute to the acquisition of the metastatic phenotype of melanoma is presented and discussed. PMID- 19308690 TI - The prometastatic microenvironment of the liver. AB - The liver is a major metastasis-susceptible site and majority of patients with hepatic metastasis die from the disease in the absence of efficient treatments. The intrahepatic circulation and microvascular arrest of cancer cells trigger a local inflammatory reaction leading to cancer cell apoptosis and cytotoxicity via oxidative stress mediators (mainly nitric oxide and hydrogen peroxide) and hepatic natural killer cells. However, certain cancer cells that resist or even deactivate these anti-tumoral defense mechanisms still can adhere to endothelial cells of the hepatic microvasculature through proinflammatory cytokine-mediated mechanisms. During their temporary residence, some of these cancer cells ignore growth-inhibitory factors while respond to proliferation-stimulating factors released from tumor-activated hepatocytes and sinusoidal cells. This leads to avascular micrometastasis generation in periportal areas of hepatic lobules. Hepatocytes and myofibroblasts derived from portal tracts and activated hepatic stellate cells are next recruited into some of these avascular micrometastases. These create a private microenvironment that supports their development through the specific release of both proangiogenic factors and cancer cell invasion- and proliferation-stimulating factors. Moreover, both soluble factors from tumor activated hepatocytes and myofibroblasts also contribute to the regulation of metastatic cancer cell genes. Therefore, the liver offers a prometastatic microenvironment to circulating cancer cells that supports metastasis development. The ability to resist anti-tumor hepatic defense and to take advantage of hepatic cell-derived factors are key phenotypic properties of liver metastasizing cancer cells. Knowledge on hepatic metastasis regulation by microenvironment opens multiple opportunities for metastasis inhibition at both subclinical and advanced stages. In addition, together with metastasis-related gene profiles revealing the existence of liver metastasis potential in primary tumors, new biomarkers on the prometastatic microenvironment of the liver may be helpful for the individual assessment of hepatic metastasis risk in cancer patients. PMID- 19308691 TI - Microenvironment changes (in pH) affect VEGF alternative splicing. AB - Vascular endothelial growth factor-A (VEGF-A) has several isoforms, which differ in their capacity to bind extracellular matrix proteins and also in their affinity for VEGF receptors. Although the relative contribution of the VEGF isoforms has been studied in tumor angiogenesis, little is known about the mechanisms that regulate the alternative splicing process. Here, we tested microenvironment cues that might regulate VEGF alternative splicing. To test this, we used endometrial cancer cells that produce all VEGF isoforms as a model, and exposed them to varying pH levels, hormones, glucose and CoCl(2) (to mimic hypoxia). Low pH had the most consistent effects in inducing variations in VEGF splicing pattern (VEGF121 increased significantly, p < 0.001, when compared to VEGF145, 165 or 189). This was accompanied by activation of the p38 stress pathway and SR proteins (splicing factors) expression and phosphorylation. SF2/ASF, SRp20 and SRp40 down-regulation by siRNA impaired the effects of pH stimulation, blocking the shift in VEGF isoforms production. Taken together, we show for the first time that acidosis (low pH) regulates VEGF-A alternative splicing, may be through p38 activation and suggest the possible SR proteins involved in this process. PMID- 19308692 TI - Sialyl Lewis X expression and lymphatic microvessel density in primary tumors of node-negative colorectal cancer patients predict disease recurrence. AB - Up to 30% of curatively resected colorectal cancer patients with tumor-negative lymph nodes, show disease recurrence. We assessed whether these high-risk patients can be identified by examining primary tumors for the following blood and lymphatic vasculature markers: A) sialyl Lewis X (sLeX), vascular endothelial growth factor (VEGF)-C and VEGF-D expression; B) blood and lymphatic microvessel density (BMVD/LMVD); and C) the presence of blood and lymphatic vessel invasion. Thirty-six cases (disease recurrence within 5 years) and 72 controls (no disease recurrence for at least 5 years) were selected in a case-control design. Tumor sections were stained by antibodies CSLEX1 (sLeX), anti-VEGF-C, anti-VEGF-D, anti CD31 (BMVD) or D2-40 (LMVD) to determine the parameters as mentioned above. A multivariate analysis showed sLeX expression and high LMVD (odds ratio 5.1, 95% confidence interval 1.3-20.0 and odds ratio 3.1, 95% confidence interval 1.0 10.0, respectively) to be independent factors predicting disease recurrence. Expression of sLeX correlated with liver metastases (P = 0.015). A high LMVD was related to regional intra-abdominal or intrapelvic metastases in lymph nodes and distant metastases other than in the liver and lungs such as peritoneum, bones, brain and adrenal glands (P = 0.004). A high BMVD in the invasive front correlated with lung metastases (P = 0.018). We show that high-risk node-negative colorectal cancer patients can be identified by primary tumor assessment for sLeX expression and LMVD. Our results are consistent with the notion that both lymphatic and hematogenous metastasis play a role in colorectal cancer. PMID- 19308693 TI - Neuroblastoma cell death is induced by inorganic arsenic trioxide (As(2)O(3)) and inhibited by a normal human bone marrow cell-derived factor. AB - Three phenotypically distinct cell types are present in human neuroblastomas (NB) and NB cell lines: I-type stem cells, N-type neuroblastic precursors, and S-type Schwannian/melanoblastic precursors. The stimulation of human N-type neuroblastoma cell proliferation by normal human bone marrow monocytic cell conditioned medium (BMCM) has been demonstrated in vitro, a finding consistent with the high frequency of bone marrow (BM) metastases in patients with advanced NB. Inorganic arsenic trioxide (As(2)O(3)), already clinically approved for the treatment of several hematological malignancies, is currently under investigation for NB. Recent studies show that As(2)O(3) induces apoptosis in NB cells. We examined the impact of BMCM on growth and survival of As(2)O(3)-treated NB cell lines, to evaluate the response of cultured NB cell variants to regulatory agents. We studied the effect of BMCM on survival and clonogenic growth of eleven As(2)O(3)-treated NB cell lines grown in sparsely seeded, non-adherent, semi solid cultures. As(2)O(3) had a strong inhibitory effect on survival of all tested NB cell lines. BMCM augmented cell growth and survival and reversed the inhibitory action of As(2)O(3) in all tested cell lines, but most strongly in N type cells(.) While As(2)O(3) effectively reduced survival of all tested NB cell lines, BMCM effectively impacted its inhibitory action. Better understanding of micro-environmental regulators affecting human NB tumor cell growth and survival may be seminal to the development of therapeutic strategies and clinically effective agents for this condition. PMID- 19308694 TI - Systems biology: a therapeutic target for tumor therapy. AB - Tumor-related activities that seem to be operationally induced by the division of function, such as inflammation, neoangiogenesis, Warburg effect, immune response, extracellular matrix remodeling, cell proliferation rate, apoptosis, coagulation effects, present itself from a systems perspective as an enhancement of complexity. We hypothesized, that tumor systems-directed therapies might have the capability to use aggregated action effects, as adjustable sizes to therapeutically modulate the tumor systems' stability, homeostasis, and robustness. We performed a retrospective analysis of recently published data on 224 patients with advanced and heavily pre-treated (10% to 63%) vascular sarcoma, melanoma, renal clear cell, cholangiocellular, carcinoma, hormone-refractory prostate cancer, and multivisceral Langerhans' cell histiocytosis enrolled in nine multi-center phase II trials (11 centers). Each patient received a multi targeted systems-directed therapy that consisted of metronomic low-dose chemotherapy, a COX-2 inhibitor, combined with one or two transcription modulators, pioglitazone +/- dexamethasone or IFN-alpha. These treatment schedules may attenuate the metastatic potential, tumor-associated inflammation, may exert site-specific activities, and induce long-term disease stabilization followed by prolonged objective response (3% to 48%) despite poor monoactivity of the respective drugs. Progression-free survival data are comparable with those of reductionist-designed standard first-line therapies. The differential response patterns indicate the therapies' systems biological activity. Understanding systems biology as adjustable size may break through the barrier of complex tumor stroma-interactions in a therapeutically relevant way: Comparatively high efficacy at moderate toxicity. Structured systems-directed therapies in metastatic cancer may get a source for detecting the topology of tumor-associated complex aggregated action effects as adjustable sizes available for targeted biomodulatory therapies. PMID- 19308695 TI - Foreword: the many fascinating functions of SMC protein complexes. PMID- 19308696 TI - Condensin: Architect of mitotic chromosomes. AB - Condensin is a highly conserved pentameric complex consisting of two structural maintenance of chromosome (SMC) ATPase subunits and three auxiliary components. While initially regarded as a key driver of mitotic chromosome condensation, condensin is increasingly viewed as having a more subtle influence on chromosome architecture. The two condensin complexes are required to direct the correct folding and organization of chromosomes prior to anaphase and for keeping the chromosomes compact as they separate to the poles. This ancient complex is essential in mitosis and meiosis and has additional roles in gene regulation and DNA repair. The wide variety of biochemical and genetic tools available are gradually unravelling the numerous roles condensin plays during the cell cycle and shedding light on its mechanism of action. PMID- 19308697 TI - Assays for mitotic chromosome condensation in live yeast and mammalian cells. AB - The dynamic reorganization of chromatin into rigid and compact mitotic chromosomes is of fundamental importance for faithful chromosome segregation. Owing to the difficulty of investigating this process under physiological conditions, the exact morphological transitions and the molecular machinery driving chromosome condensation remain poorly defined. Here, we review how imaging-based methods can be used to quantitate chromosome condensation in vivo, focusing on yeast and animal tissue culture cells as widely used model systems. We discuss approaches how to address structural dynamics of condensing chromosomes and chromosome segments, as well as to probe for mechanical properties of mitotic chromosomes. Application of such methods to systematic perturbation studies will provide a means to reveal the molecular networks underlying the regulation of mitotic chromosome condensation. PMID- 19308698 TI - Intersection of ChIP and FLIP, genomic methods to study the dynamics of the cohesin proteins. AB - The evolutionarily conserved cohesin proteins Smc1, Smc3, Rad21 (Mcd1), and Scc3 function in the cohesin complex that provides the basis for chromosome cohesion and is involved in gene regulation. Understanding how these proteins link together the genome requires the use of whole-genome approaches to study the molecular mechanisms of these essential proteins. While chromatin immunoprecipitation followed by DNA microarray (ChIP-chip) studies have provided a snapshot in time of where these proteins associate with various genomes, the cohesin proteins are dynamic in their localization and interactions on chromatin. Study of the dynamic nature of these proteins requires approaches such as live cell imaging. We present evidence from fluorescence loss in photobleaching (FLIP) experiments in budding yeast that the decay constant of each cohesin subunit is approximately 60-90 s in interphase. The decay constant on chromatin increases from G(1) to S phase to metaphase, consistent with the interaction with chromatin becoming more stable once chromosomes are cohered. A small population of Smc3 at a position consistent with centromeric location has a longer decay constant than bulk Smc3. The characterization of the interaction of cohesin with chromatin, in terms of both its position and its dynamics, may be key to understanding how this protein complex contributes to chromosome segregation and gene regulation. PMID- 19308699 TI - Lampbrush chromosomes enable study of cohesin dynamics. AB - The lampbrush chromosomes present in the nuclei of amphibian oocytes offer unique biological approaches for study of the mechanisms that regulate chromatin structure with high spatial resolution. We discuss fundamental aspects of the remarkable organization and plasticity exhibited by lampbrush chromosomes. We then utilize lampbrush chromosomes to characterize the chromosomal distribution and dynamics of cohesin, the four-protein complex (RAD21/MCD1/SCC1, SMC1, SMC3, SCC3/SA2) responsible for sister chromatid cohesion. We find that endogenous SMC3 and newly expressed hRAD21 co-localize on chromosomal axes, sites where sister chromatids are tightly paired. We present evidence suggesting that hRAD21 recruitment to lampbrush chromosomes is modulated by chromosomal SMC1 and SMC3. Notably, using a technique for de novo chromosome assembly, we demonstrate that both SMC3 and hRAD21 are recruited to single, unreplicated lampbrush chromatids. Finally, we used our novel method of analyzing the oocyte nucleus under oil combined with fluorescence recovery after photobleaching, to provide direct evidence that cohesin is highly dynamic at discrete, condensed chromosomal regions. Collectively, these data demonstrate that lampbrush chromosomes provide a unique and powerful tool for combining biochemical and cytological analyses for dissection of complex chromosomal processes. PMID- 19308700 TI - Cohesin, gene expression and development: lessons from Drosophila. AB - The cohesin complex, discovered through its role in sister chromatid cohesion, also plays roles in gene expression and development in organisms from yeast to human. This review highlights what has been learned about the gene control and developmental functions of cohesin and the Nipped-B (NIPBL/Scc2) cohesin loading factor in Drosophila. The Drosophila studies have provided unique insights into the aetiology of Cornelia de Lange syndrome (CdLS), which is caused by mutations affecting sister chromatid cohesion proteins in humans. In vivo experiments with Drosophila show that cohesin and Nipped-B have dosage-sensitive effects on the functions of many evolutionarily conserved genes and developmental pathways. Genome-wide studies with Drosophila cultured cells show that Nipped-B and cohesin co-localize on chromosomes, and bind preferentially, but not exclusively, to many actively transcribed genes and their regulatory sequences, including many of the proposed in vivo target genes. In contrast, the cohesion factors are largely excluded from genes silenced by Polycomb group (PcG) proteins. Combined, the in vivo genetic data and the binding patterns of cohesin and Nipped-B in cultured cells are consistent with the hypothesis that they control the action of gene regulatory sequences, including transcriptional enhancers and insulators, and suggest that they might also help define active chromatin domains and influence transcriptional elongation. PMID- 19308701 TI - How cohesin and CTCF cooperate in regulating gene expression. AB - Cohesin is a DNA-binding protein complex that is essential for sister chromatid cohesion and facilitates the repair of damaged DNA. In addition, cohesin has important roles in regulating gene expression, but the molecular mechanisms of this function are poorly understood. Recent experiments have revealed that cohesin binds to the same sites in mammalian genomes as the zinc finger transcription factor CTCF. At a few loci CTCF has been shown to function as an enhancer-blocking transcriptional insulator, and recent observations indicate that this function depends on cohesin. Here we review what is known about the roles of cohesin and CTCF in regulating gene expression in mammalian cells, and we discuss how cohesin might mediate the insulator function of CTCF. PMID- 19308702 TI - C. elegans dosage compensation: a window into mechanisms of domain-scale gene regulation. AB - The C. elegans dosage compensation complex (DCC) reduces transcript levels from each of the two hermaphrodite X chromosomes to equalize X-linked gene expression to that of XO males. Several of the proteins that comprise the DCC are homologous to subunits of the evolutionarily conserved condensin complexes, which in most organisms function in mitotic and meiotic chromosome condensation. These include the DCC subunits MIX-1 and DPY-27, which belong to the structural maintenance of chromosomes (SMC) family of proteins. Several of the C. elegans DCC subunits also perform double duty as members of the canonical meiotic and mitotic condensin complexes. Here, we review what is known about the C. elegans DCC and how study of this model might shed light on general mechanisms of domain-scale transcriptional regulation. We discuss how condensin-like complexes may be targeted to specific chromosomal locations for performance of their functions. PMID- 19308703 TI - Heterochromatin and the cohesion of sister chromatids. AB - Heterochromatin, once thought to be the useless junk of chromosomes, is now known to play significant roles in biology. Underlying much of this newfound fame are links between the repressive chromatin structure and cohesin, the protein complex that mediates sister chromatid cohesion. Heterochromatin-mediated recruitment and retention of cohesin to domains flanking centromeres promotes proper attachment of chromosomes to the mitotic and meiotic spindles. Heterochromatin assembled periodically between convergently transcribed genes also recruits cohesin, which promotes a novel form of transcription termination. Heterochromatin-like structures in budding yeast also recruit cohesin. Here the complex appears to regulate transcriptional silencing and recombination between repeated DNA sequences. The link between heterochromatin and cohesin is particularly relevant to human health. In Roberts-SC phocomelia syndrome, heterochromatic cohesion is selectively lost due to mutation of the acetyltransferase responsible for cohesin activation. In this review I discuss recent work that relates to these relationships between heterochromatin and cohesin. PMID- 19308704 TI - Studies of meiosis disclose distinct roles of cohesion in the core centromere and pericentromeric regions. AB - During meiosis, a single round of genome duplication is followed by two sequential rounds of chromosome segregation. Through this process, a diploid parent cell generates gametes with a haploid set of chromosomes. A characteristic of meiotic chromosome segregation is a stepwise loss of sister chromatid cohesion along chromosomal arms and at centromeres. Whereas arm cohesion plays an important role in ensuring homologue disjunction at meiosis I, persisting cohesion at pericentromeric regions throughout meiosis I is essential for the faithful equational segregation of sisters in the following meiosis II, similar to mitosis. A widely conserved pericentromeric protein called shugoshin, which associates with protein phosphatase 2A (PP2A), plays a critical role in this protection of cohesin. Another key aspect of meiosis I is the establishment of monopolar attachment of sister kinetochores to spindle microtubules. Cohesion or physical linkage at the core centromeres, where kinetochores assemble, may conjoin sister kinetochores, leading to monopolar attachment. A meiosis-specific kinetochore factor such as fission yeast Moa1 or budding yeast monopolin contributes to this regulation. We propose that cohesion at the core centromere and pericentromeric regions plays distinct roles, especially in defining the orientation of kinetochores. PMID- 19308705 TI - The unnamed complex: what do we know about Smc5-Smc6? AB - The structural maintenance of chromosome (SMC) proteins constitute the cores of three protein complexes involved in chromosome metabolism; cohesin, condensin and the Smc5-Smc6 complex. While the roles of cohesin and condensin in sister chromatid cohesion and chromosome condensation respectively have been described, the cellular function of Smc5-Smc6 is as yet not understood, consequently the less descriptive name. The complex is involved in a variety of DNA repair pathways. It contains activities reminiscent of those described for cohesin and condensin, as well as several DNA helicases and endonucleases. It is required for sister chromatid recombination, and smc5-smc6 mutants suffer from the accumulation of unscheduled recombination intermediates. The complex contains a SUMO-ligase and potentially an ubiquitin-ligase; thus Smc5-Smc6 might presently have a dull name, but it seems destined to be recognized as a key player in the maintenance of chromosome stability. In this review we summarize our present understanding of this enigmatic protein complex. PMID- 19308706 TI - Dynamics of the bacterial SMC complex and SMC-like proteins involved in DNA repair. AB - Bacteria and archaea possess several different SMC-like proteins, which perform essential functions in a variety of chromosome dynamics, such as chromosome compaction, segregation, and DNA repair. SMC-like proteins localize to distinct sites within the cells at different time points in the cell cycle, or are recruited to sites of DNA breaks and damage. The bacterial SMC (MukB) complex appears to perform a condensin-like function, while SbcC and RecN act early during DNA repair, but apparently at different sites within the cells. Thus, bacterial SMC-like proteins have dynamic functions in chromosome segregation and maintenance of genetic stability. PMID- 19308707 TI - RAD50, an SMC family member with multiple roles in DNA break repair: how does ATP affect function? AB - The protein complex including Mre11, Rad50, and Nbs1 (MRN) functions in DNA double-strand break repair to recognize and process DNA ends as well as signal for cell cycle arrest. Amino acid sequence similarity and overall architecture make Rad50 a member of the structural maintenance of chromosome (SMC) protein family. Like SMC proteins, Rad50 function depends on ATP binding and hydrolysis. All current evidence indicates that ATP binding and hydrolysis cause architectural rearrangements in SMC protein complexes that are important for their functions in organizing DNA. In the case of the MRN complex, the functional significance of ATP binding and hydrolysis are not yet defined. Here we review the data on the ATP-dependent activities of MRN and their possible mechanistic significance. We present some speculation on the role of ATP for function of the MRN complex based on the similarities and differences in the molecular architecture of the Rad50-containing complexes and the SMC complexes condensin and cohesin. PMID- 19308708 TI - A participatory workplace intervention for employees with distress and lost time: a feasibility evaluation within a randomized controlled trial. AB - INTRODUCTION: Little is known about feasibility and acceptability of return to work (RTW) interventions for mental health problems. RTW for mental health problems is more complicated than for musculoskeletal problems due to stigmatization at the workplace. A participatory workplace intervention was developed in which an employee and supervisor identify and prioritize obstacles and solutions for RTW guided by a RTW coordinator. This paper is a feasibility study of this innovative intervention for employees with distress. The aims of this study were to describe the reach and extent of implementation of the workplace intervention, the satisfaction and expectations of all stakeholders, and the intention to use the workplace intervention in the future. METHODS: Eligible for this study were employees who had been on sick leave from regular work for 2-8 weeks with distress. Data were collected from the employees, their supervisors, RTW coordinators, and occupational physicians by means of standardized matrices and questionnaires at baseline and 3 months follow-up. Reach, implementation, satisfaction, expectations, and maintenance regarding the workplace intervention were described. RESULTS: Of the 56 employees with distress eligible to receive the workplace intervention, 40 employees, their supervisors and RTW coordinators actually participated in the intervention. They identified 151 obstacles for RTW mostly related to job design, communication, mental workload and person-related stress factors. The 281 consensus-based solutions identified were mostly related to job design, communication and training. Of those solutions, 72% was realized at the evaluation with the employee and supervisor. Overall, employees, supervisors and occupational health professionals were satisfied with the workplace intervention and occupational health professionals rated it with a 7.1. Time-investment was the only barrier for implementation reported by the occupational health professionals. CONCLUSIONS: The results of this study indicate a high feasibility for a broad implementation of a participatory workplace intervention for employees with distress and lost time, and their supervisors. PMID- 19308709 TI - Factors that promote or hinder young disabled people in work participation: a systematic review. AB - INTRODUCTION: The aim of this systematic review was to study factors which promote or hinder young disabled people entering the labor market. METHODS: We systematically searched PubMed (by means of MESH and text words), EMBASE, PsycINFO, Web of Science and CINAHL for studies regarding (1) disabled patients diagnosed before the age of 18 years and (2) factors of work participation. RESULTS: Out of 1,268 retrieved studies and 28 extended studies from references and four from experts, ten articles were included. Promoting factors are male gender, high educational level, age at survey, low depression scores, high dispositional optimism and high psychosocial functioning. Female and low educational level gives high odds of unemployment just like low IQ, inpatient treatment during follow up, epilepsy, motor impairment, wheelchair dependency, functional limitations, co-morbidity, physical disability and chronic health conditions combined with mental retardation. High dose cranial radiotherapy, type of cancer, and age of diagnosis also interfered with employment. CONCLUSIONS: Of the promoting factors, education appeared to be important, and several physical obstructions were found to be hindering factors. The last mentioned factors can be influenced in contrast to for instance age and gender. However, to optimize work participation of this group of young disabled it is important to know the promoting or hindering influence for employment. PMID- 19308710 TI - Clinical and molecular characteristics of 35 Chinese children with Wiskott Aldrich syndrome. AB - BACKGROUND: Wiskott-Aldrich syndrome (WAS) is a rare primary immunodeficiency disease, with an incidence of 4/1,000,000 live male births. In China, an estimated number of 35 babies with WAS are born each year, but likely many remain undiagnosed. OBJECTIVES: The objectives of study were to review the clinical and molecular characteristics of a cohort of Chinese children with WAS and to describe the long-term outcome of those who underwent hematopoietic stem cell transplant (HSCT). MATERIALS AND METHOD: Records of 35 patients diagnosed with WAS during 1991-2008 were reviewed. Genetic diagnosis was established by direct gene sequencing. RESULTS: All patients had classical WAS phenotype. WASP mutations were identified in 33 patients from 29 families. Nine patients underwent HSCT at a mean age of 22.1 months (match-unrelated donor, n = 5; mismatched related donor, n = 2; matched-sibling donor, n = 2). Post-transplant immune hemolytic anemia and thrombocytopenia occurred in three patients with complete resolution. All patients survived without significant long-term complications and had full platelet, T and B lymphocyte recovery within 2 years post-transplant. CONCLUSION: In the past decade, there has been significant improvement in clinical and genetic diagnosis of WAS in Chinese. We demonstrated excellent long-term survival in patients who underwent HSCT. Early workup for transplant should be advocated for children with classical WAS before they suffer from major disease complications and morbidities. PMID- 19308711 TI - Birth weight effects on children's mental, motor, and physical development: evidence from twins data. AB - OBJECTIVE: To determine the effect of very low birth weight (VLBW; <1500 g) and moderately low birth weight (MLBW; 1500-2499 g) on children's mental and motor development and physical growth during the first 2 years of life and whether VLBW and MLBW babies catch up to normal birth weight (NBW; > or =2500 g) children by age 2. METHODS: We use data on dizygotic (DZ) and monozygotic (MZ) twins and singleton births from the first two waves of the Early Childhood Longitudinal Study-Birth Cohort (ECLS-B), a nationally representative dataset of children born in the US in 2001. We estimate the effects of VLBW and MLBW on children's mental and motor development scores, weight-for-age, weight-for-length, weight-for height, and length-for-age z-scores at 9 months and 2 years. We examine whether differences in outcomes within twin pairs are related to differences in their birth weights. The within-twins analysis is conducted on samples of DZ and MZ twins. For comparison, we also estimate birth weight effects on child outcomes from multivariate linear regression models using the full singleton and twins' sample. We also estimate the effect of being small-for-gestational age (SGA; birth weight <10th percentile for gestation) using the same set of models in order to separate out the effects of fetal growth restriction from prematurity. RESULTS: Evaluation of all births showed that VLBW and MLBW have large negative effects on mental development, motor development, and growth at 9 months and 2 years of age. However, results from within-twin models with DZ twins that control for shared maternal and environmental factors showed much less effect of birth weight on mental or motor development, but continued large effects on growth for the VLBW group. Within-twin models with MZ twins that control for shared maternal, environmental, and genetic factors showed statistically insignificant effects of birth weight on mental and motor development, but continued effects on growth. Similar patterns were found when examining the effects of SGA. CONCLUSIONS: After controlling for the influence of maternal, environmental, and genetic factors, low birth weight has at most a small negative effect on children's mental and motor development in their first 2 years of life. However, low birth weight is a major risk factor for children's physical growth in the early years and there is no evidence of catch-up by age 2. PMID- 19308712 TI - Mammillary body atrophy in acute liver failure and acute-on-chronic liver failure of nonalcoholic etiology. AB - The most common cause of atrophy of mammillary bodies (MBs) is thiamine deficiency, which is very common in patients with alcoholic liver disease. The purpose of this study was to look for changes in MBs using brain magnetic resonance imaging (MRI) in patients with acute liver failure (ALF), acute-on chronic liver failure (ACLF) and chronic liver failure (CLF) of non-alcoholic etiology. Volumes of MBs and caudate nuclei (CNs) were quantified in nine patients with ALF, 17 with ACLF, 18 with CLF and in 24 healthy controls. Volume of these structures was quantified again three weeks after clinical recovery in five patients with ALF who had survived their illness. Volume of left, right and both MBs was significantly decreased (p < 0.05) in patients with ALF and ACLF whereas there was no change in patients with CLF, when compared with healthy controls. However CN volumes did not change significantly compared to controls in any of the three patient groups. In the follow-up study significant recovery in volume of MBs was noted compared to baseline values in the ALF patients. We conclude that significant volume loss occurs in MBs in patients with ALF and ACLF of non-alcoholic etiology but not in CLF. This loss of MBs volume recovers substantially in patients with ALF who survive their illness. PMID- 19308713 TI - Codon volatility of hemagglutinin genes of H5N1 avian influenza viruses from different clades. AB - Codon volatility is a method recently developed to estimate selective pressures on proteins on the basis of their synonymous codon usage. Volatility of a codon was defined as the fraction of single nucleotide substitutions that would be nonsynonymous. Higher volatility may indicate that the gene has been under more positive selection in the recent past. We analyzed volatility of hemagglutinin genes of H5N1 viruses in the recent outbreaks and observed differences in the volatility among viruses of different clades. The codon volatility of subclade 2.1 viruses from Indonesia was the lowest among all H5N1 clades and subclades. Time series analyses since the beginning of the epidemic in 2004 showed that codon volatility of subclade 2.1 has gradually decreased, while those of other major clades have been increasing. This may reflect differences in the recent evolution of these viruses. PMID- 19308714 TI - Negotiating sex and sexualities: the use of sexual tags in the Brazilian sex trade workplace. AB - Based on data from participant observation and interviews with Brazilian sex workers and their clients, I examined the intersections of sexual and gender identities with sexual behaviors. Specifically, I was interested in how sex workers managed and negotiated their sexualities in the workplace. First, I described the layout and the social actors at a popular termas in a major Brazilian city. Then, I examined the interactions in the termas and how Brazilian sex workers use sexual identity terms to describe themselves. I argued that the use of these terms do not necessarily indicate sex workers' adoption of that sexual identity. Critical to them are the symbolic presentation of gender identity and the presentation of self at the work place. I developed a theoretical construct, sexual tags, which contributes to the literature by suggesting that sexual identity is situational and, more importantly, that the terms may not mean as much to the individuals or to be consistent with our understanding of what they ought to imply, especially when individuals are importing western terms to relate to their foreign clients. While using sexual tags does not preclude sex workers from eventually adopting that identity, it could also mean that some sex workers are claiming a sexual identity simply for strategic purposes. Another possible scenario is that sexual tags reflect the diverse interpretations of sexuality because of the available choices created by the situational manipulation of sexual meanings within a local context. PMID- 19308715 TI - Sexual compulsivity and sexual risk in gay and bisexual men. AB - Much of our understanding of the association between the Sexual Compulsivity Scale (SCS) and sexual risk behavior among men who have sex with men (MSM) has been limited to samples of HIV positive MSM only. Using data from a community based survey of gay and bisexual men (n = 1214), this analysis sought to further evaluate the association between the SCS and sexual risk behavior. The SCS was significantly associated with a variety of sexual risk behaviors, including having sex under the influence of club drugs, engaging in unprotected anal sex (receptive or insertive) with partners of the same and/or different HIV serostatus, identity as a barebacker, intentions to have bareback sex, number of recent sex partners, and temptation for unsafe sex. The SCS was also significantly associated with having engaged in a variety of specialized sexual behaviors (i.e., fetishes), many of which can increase HIV transmission risks. Finally, in multivariate analyses, the SCS significantly predicted unprotected sex with a non-main partner even when controlling for race, HIV serostatus, age, identity as a barebacker, and club drug use. These data indicate that the SCS may be able to serve as an indicator to detect HIV-associated sexual risk behavior in community-based samples of gay and bisexual men. PMID- 19308716 TI - A longitudinal investigation of coping strategies and quality of life among younger women with breast cancer. AB - It is generally assumed that coping strategies impact quality of life (QOL). It is plausible that QOL determines use of coping strategies. This research examines coping strategies over time and the reciprocal relationship between coping strategies and QOL among younger women with breast cancer. Women with breast cancer (N = 267; mean age = 43 years) completed surveys within 6 months of diagnosis and 6 weeks and 6 months later. Surveys included questions on coping strategies, QOL, medical factors, and sociodemographics. Positive cognitive restructuring was the most frequently used strategy. Over time, use of seeking social support, spirituality, and wishful thinking declined, while detachment increased. Prior QOL predicted three subsequent coping strategies (seeking social support, keeping feelings to self, wishful thinking). Coping strategies were minimally related to subsequent QOL. Coping strategies and QOL are dynamic processes. QOL may predict coping strategies equally or more than vice versa. PMID- 19308717 TI - Is sexual serosorting occurring among HIV-positive injection drug users? Comparison between those with HIV-positive partners only, HIV-negative partners only, and those with any partners of unknown status. AB - Using baseline data from a multi-site, randomized controlled study (INSPIRE), we categorized 999 HIV-positive IDUs into three groups based on serostatus of their sex partners. Our data provide some evidence for serosorting occurring in our sample; about 40% of the sample had sex exclusively with HIV-positive partners, and about half of them reported having unprotected sex with these partners. Twenty per cent had sex exclusively with HIV-negative partners; their sexual behaviors tended to be least risky with about two-thirds reporting their sex was protected. However, we also found that another 40% had at least one partner of unknown HIV status and sexual and drug risk was the highest among them. They were also least empowered, showing attributes that may undermine HIV prevention. Some of these findings are consistent with findings from MSM studies, suggesting that partner selection practices are similar between primarily heterosexual IDUs and MSM. PMID- 19308718 TI - Fertility intentions of HIV-1 infected and uninfected women in Malawi: a longitudinal study. AB - This study aimed to determine changes in fertility intentions of HIV-1 infected and uninfected reproductive age women in Blantyre, Malawi. Participants were asked about their fertility intentions at baseline and at 3-month visits for 1 year. Time-to-event statistical models were used to determine factors associated with changes in fertility intentions. Overall, 842 HIV uninfected and 844 HIV infected women were enrolled. The hazard of changing from wanting no more children at baseline to wanting more children at follow-up was 61% lower among HIV infected women compared to HIV uninfected women (P < 0.01) after adjusting for other factors, while HIV infected women were approximately 3 times more likely to change to wanting no more children. The overall pregnancy rate after 12 months was 14.9 per 100 person-years and did not differ among 102 HIV uninfected and 100 infected women who became pregnant. HIV infection is a significant predictor of fertility intentions over time. PMID- 19308719 TI - A qualitative assessment of decisions affecting contraceptive utilization and fertility intentions among HIV-positive women in Soweto, South Africa. AB - The HIV epidemic in sub-Saharan Africa disproportionately affects women of reproductive age. The increasing provision of Highly Active Anti-Retroviral Therapy (HAART) with improved prognosis and maternal-fetal outcomes calls for an understanding of fertility planning for HIV-positive women. We describe the effect of HIV and HAART on pregnancy desires and contraceptive use among HIV positive women in Soweto, South Africa. Focus group discussions and in-depth interviews were conducted with 42 HIV-positive women of reproductive age. Analysis was performed using ATLAS-ti (ATLAS-ti Center, Berlin). Emergent themes were impact of HIV diagnosis on pregnancy intentions; factors affecting contraceptive uptake including real and normative side effects, body image, and perceived vaginal wetness; and the mitigating influence of partnership on both pregnancy intentions and contraceptive use. Routine counseling about pregnancy desires and contraception should be offered to HIV-positive women. PMID- 19308720 TI - Male involvement in PMTCT services in Mbeya Region, Tanzania. AB - Throughout all stages of programmes for the prevention of mother-to-child transmission of HIV (PMTCT), high dropout rates are common. Increased male involvement and couples' joint HIV counselling/testing during antenatal care (ANC) seem crucial for improving PMTCT outcomes. Our study assessed male attitudes regarding partner involvement into ANC/PMTCT services in Mbeya Region, Tanzania, conducting 124 individual interviews and six focus group discussions. Almost all respondents generally supported PMTCT interventions. Mentioned barriers to ANC/PMTCT attendance included lacking information/knowledge, no time, neglected importance, the services representing a female responsibility, or fear of HIV-test results. Only few perceived couple HIV counselling/testing as disadvantageous. Among fathers who had refused previous ANC/PMTCT attendance, most had done so even though they were not perceiving a disadvantage about couple counselling/testing. The contradiction between men's beneficial attitudes towards their involvement and low participation rates suggests that external barriers play a large role in this decision-making process and that partner's needs should be more specifically addressed in ANC/PMTCT services. PMID- 19308721 TI - HIV transmission potential among local and migrant factory workers in Kolkata, India. AB - Migrant workers in India play a key role in the spread of HIV. Kolkata is a common destination for workers, who may acquire infection and transmit it to their wives and/or other sexual partners. We investigated sexual relations and condom use by factory workers. Migrant and local factory workers were randomly selected from five wards of Kolkata. Information was collected about demographic and socio-economic characteristics, sexual relationships, condom usage, and perceptions and intent to use condoms. Condom use was very low in both groups of workers, particularly among migrants. Many married workers visited female sex workers but never used condoms. Few intended to use condoms, and if they did, it did not always translate into actual usage. There is great potential for transmission of HIV/sexually transmitted infections by these workers. Carefully designed intervention and education programs in the context of low literacy and cultural norms are urgently needed. PMID- 19308722 TI - Pilot trial of an intervention aimed at modifying drug preparation practices among injection drug users in Puerto Rico. AB - Injection drug users (IDUs) contaminate preparation materials with blood-borne pathogens by using syringes as measuring and dispensing devices. In collaboration with IDUs, we developed a preventive intervention consisting of four new preparation practices aimed at avoiding the use of syringes in the preparation, and reducing the contamination of the materials. This report describes the results of a pilot trial introducing the new practices to ascertain their adoption potential and their potential efficacy in reducing contamination. Participants comprised 37 active IDUs among whom the new practices were promoted during 16 weeks. In addition to self-reported behaviors, the study collected cookers and plastic caps from shooting galleries and tested them for the presence of blood residues. Adoption rates were: (1) cleaning of skin area with hand sanitizer--65.6%; (2) directly pouring water with a dropper into the cooker- 56.3%; (3) drawing drug solution with a preparation syringe and syringe filter- 34.4%; and, (4) backload rinsing syringes--53.1%. Rates of blood residues detected in cookers and plastic caps were 41.7% prior to the trial, 28.6% at week 8, 24.6% at week 14, and 12.0% at week 18. We believe the results of the pilot trial are compelling and suggest that this intervention merits further formal testing. PMID- 19308723 TI - Comorbid problems in ADHD: degree of association, shared endophenotypes, and formation of distinct subtypes. Implications for a future DSM. AB - We aimed to assess which comorbid problems (oppositional defiant behaviors, anxiety, autistic traits, motor coordination problems, and reading problems) were most associated with Attention-Deficit/Hyperactivity Disorder (ADHD); to determine whether these comorbid problems shared executive and motor problems on an endophenotype level with ADHD; and to determine whether executive functioning (EF)-and motor-endophenotypes supported the hypothesis that ADHD with comorbid problems is a qualitatively different phenotype than ADHD without comorbid problems. An EF-and a motor-endophenotype were formed based on nine neuropsychological tasks administered to 816 children from ADHD-and control families. Additional data on comorbid problems were gathered using questionnaires. Results indicated that oppositional defiant behaviors appeared the most important comorbid problems of ADHD, followed by autistic traits, and than followed by motor coordination problems, anxiety, and reading problems. Both the EF-and motor-endophenotype were correlated and cross-correlated in siblings to autistic traits, motor coordination problems and reading problems, suggesting ADHD and these comorbid problems may possibly share familial/genetic EF and motor deficits. No such results were found for oppositional defiant behaviors and anxiety. ADHD in co-occurrence with comorbid problems may not be best seen as a distinct subtype of ADHD, but further research is warranted. PMID- 19308724 TI - How people can benefit from mental health consumer-run organizations. AB - The goal of this study is to develop a more comprehensive theoretical understanding of the processes by which people can benefit from mental health consumer-run organizations (CROs). To accomplish this goal, the concept of roles is used to create a preliminary framework that draws connections between several established theoretical explanations. To ground theory development in empirical data, 194 CRO members from 20 CROs answered open-ended questions about what personal changes occurred as a result of their CRO involvement and what CRO participation experiences enabled personal change. Data analysis led to the identification of 18 personal change categories and 7 experiences that led to change. These categories were integrated into the preliminary theoretical framework, which needed to be extended to accommodate all categories. While inevitably tentative, the final conceptualization provides a more comprehensive understanding of the processes by which people can benefit from CRO participation. PMID- 19308725 TI - The "weight" of words on the forearms during relaxation. AB - In this paper we examined the influence of repetition of weight-related sentences on the involuntary pressure forces of the forearms, when in a relaxed state. These forces were involuntary oscillations, exerted by muscle movements of the breathing-cycle and muscle movements of the arm on force sensors. We constructed a linguistic bio-mechanical system (Ablasmi), where in each arm of a padded relaxation chair contained special sensors that were specifically designed to detect the pressure forces of each forearm while participants listened to the specific recorded weight-related sentences. In this experiment we used some classic sentences, such as "your right/left arm is heavy," inspired by Autogenic Training (Schultz in Le training autogene. Paris, PUF, 1974) and some sentences, such as "your right/left arm is made of lead," inspired by Grossmann's relaxation technique. We observed that when the recorded sentences were directed to the right arm there was a significant increase of involuntary pressure forces on the right forearm. Additionally, we observed the same effect on the left forearm for the sentences directed to the left forearm. Thus, we believe we have provided experimental evidence of a definite linguistic effect of weight-related sentences on the involuntary pressure forces of the forearms. The implications of these findings are discussed. PMID- 19308726 TI - Expression of estrogenicity genes in a lineage cell culture model of human breast cancer progression. AB - TaqMan Gene Expression assays were used to profile the mRNA expression of estrogen receptor (ERalpha and ERbeta) and estrogen metabolism enzymes including cytosolic sulfotransferases (SULT1E1, SULT1A1, SULT2A1, and SULT2B1), steroid sulfatase (STS), aromatase (CYP19), 17beta-hydroxysteroid dehydrogenases (17betaHSD1 and 2), CYP1B1, and catechol-O-methyltransferase (COMT) in an MCF10A derived lineage cell culture model for basal-like human breast cancer progression and in ERalpha-positive luminal MCF7 breast cancer cells. Low levels of ERalpha and ERbeta mRNA were present in MCF10A-derived cell lines. SULT1E1 mRNA was more abundant in confluent relative to subconfluent MCF10A cells, a non-tumorigenic proliferative breast disease cell line. SULT1E1 was also expressed in preneoplastic MCF10AT1 and MCF10AT1K.cl2 cells, but was markedly repressed in neoplastic MCF10A-derived cell lines as well as in MCF7 cells. Steroid metabolizing enzymes SULT1A1 and SULT2B1 were only expressed in MCF7 cells. STS and COMT were widely detected across cell lines. Pro-estrogenic 17betaHSD1 mRNA was most abundant in neoplastic MCF10CA1a and MCF10DCIS.com cells, while 17betaHSD2 mRNA was more prominent in parental MCF10A cells. CYP1B1 mRNA was most abundant in MCF7 cells. Treatment with the histone deacetylase inhibitor trichostatin A (TSA) induced SULT1E1 and CYP19 mRNA but suppressed CYP1B1, STS, COMT, 17betaHSD1, and 17betaHSD2 mRNA in MCF10A lineage cell lines. In MCF7 cells, TSA treatment suppressed ERalpha, CYP1B1, STS, COMT, SULT1A1, and SULT2B1 but induced ERbeta, CYP19 and SULT2A1 mRNA expression. The results indicate that relative to the MCF7 breast cancer cell line, key determinants of breast estrogen metabolism are differentially regulated in the MCF10A-derived lineage model for breast cancer progression. PMID- 19308727 TI - Effect of denosumab on bone mineral density in women receiving adjuvant aromatase inhibitors for non-metastatic breast cancer: subgroup analyses of a phase 3 study. AB - Denosumab increased lumbar spine bone mineral density (BMD) versus placebo in a 2 year, randomized, placebo-controlled, phase 3 study of patients with hormone receptor-positive, non-metastatic breast cancer and low bone mass who were receiving adjuvant aromatase inhibitor therapy. In subgroup analyses at 12 and 24 months, we evaluated factors (duration and type of aromatase inhibitor, tamoxifen use, age, time since menopause, body mass index, T-score) that might influence BMD at the lumbar spine, total hip, femoral neck, and 1/3 radius. Patients were randomized to receive placebo (n = 125) or 60 mg denosumab (n = 127) subcutaneously every 6 months. In all subgroups, 12 or 24 months' treatment with denosumab was associated with larger BMD gains than placebo across multiple skeletal sites. Most increases were statistically significant (P < 0.05). Twice yearly administration of denosumab, regardless of patient subgroup or skeletal site, resulted in consistent increases in BMD versus placebo at 12 and 24 months. PMID- 19308728 TI - Economic aspects of peer support groups for psychosis. AB - Peer support groups are rarely available for patients with psychosis, despite potential clinical and economic advantages of such groups. In this study, 106 patients with psychosis were randomly allocated to minimally guided peer support in addition to care as usual (CAU), or CAU only. No relevant differences between mean total costs of both groups were found, nor were there significant differences in WHOQoL-Bref outcomes. Intervention adherence had a substantial impact on the results. It was concluded that minimally guided peer support groups for psychosis do not seem to affect overall healthcare expenses. Positive results of additional outcomes, including a significant increase in social contacts and esteem support, favour the wider implementation of such groups. PMID- 19308729 TI - Changes in treatment content of services during trauma-informed integrated services for women with co-occurring disorders. AB - The experience of trauma is highly prevalent in the lives of women with mental health and substance abuse problems. We examined how an intervention targeted to provide trauma-informed integrated services in the treatment of co-occurring disorders has changed the content of services reported by clients. We found that the intervention led to an increased provision of integrated services as well as services addressing each content area: trauma, mental health and substance abuse. There was no increase in service quantity from the intervention. Incorporation of trauma-specific element in the treatment of mental health and substance abuse may have been successfully implemented at the service level thereby better serve women with complex behavioral health histories. PMID- 19308730 TI - PRKCSH genetic mutation was not found in Taiwanese patients with polycystic liver disease. AB - Polycystic liver disease (PCLD) without polycystic kidney is infrequent in clinical setting. Family clustering is found in patients with PCLD, and it is inherited in an autosomal dominant fashion. Through positional cloning in North America and Europe (mostly in Dutch and Finnish descents), mutations in PRKCSH gene on chromosome 19 were found to be responsible for the disease. We investigated the prevalence of liver cysts and PCLD in Taiwan and investigated whether the PRKCSH mutations exist in Taiwanese. The prevalence of liver cysts is only 0.17% in people under 30 years old and increased gradually to 14.29% in people between 55 and 60 years old and 14.19% in people over 65 years old. PCLD was not found in people under 40 years old. The prevalence is 0.15% between 40 and 45 years old, and increased to 1.37% between 55 and 60 years old, 1.21% between 60 and 65 years old, and 0.99% over 65 years old. There is only one polymorphism (deletion of one GAG repeat in exon 11) found, and the genotype and allele frequency were the same in Taiwanese patients and controls. No mutation, even polymorphism reported in the literature, was found in the 20 cases of PCLD. Our results suggest that PRKCSH gene is not a major genetic cause of PCLD and there may be at least another locus responsible for the disease in Taiwan. PMID- 19308731 TI - Ethics and the compensation of immigrant workers for work-related injuries and illnesses. AB - This paper examines the compensation process for work-related injuries and illnesses by assessing the trajectories of a sample of immigrant and non immigrant workers (n = 104) in Montreal. Workers were interviewed to analyze the complexity associated with the compensation process. Experts specialized in compensation issues assessed the difficulty of the interviewees' compensation process. Immigrant workers faced greater difficulties with medical, legal, and administrative issues than non-immigrants did. While immigrant workers' claim forms tended to be written more often by employers or friends (58% vs. 8%), the claims were still more often contested by employers (64% vs. 24%). Immigrant workers were less likely to obtain a precise diagnosis (64% vs. 42%) and upon returning to work were more likely to face sub-optimal conditions. Such results throw into relief issues of ethics and equity in host societies that are building their economy with migrant workers. PMID- 19308732 TI - Continuous left ventricular ejection fraction monitoring by aortic pressure waveform analysis. AB - We developed a technique to monitor left ventricular ejection fraction (EF) by model-based analysis of the aortic pressure waveform. First, the aortic pressure waveform is represented with a lumped parameter circulatory model. Then, the model is fitted to each beat of the waveform to estimate its lumped parameters to within a constant scale factor equal to the arterial compliance (C (a)). Finally, the proportional parameter estimates are utilized to compute beat-to-beat absolute EF by cancelation of the C (a) scale factor. In this way, in contrast to conventional imaging, EF may be continuously monitored without any ventricular geometry assumptions. Moreover, with the proportional parameter estimates, relative changes in beat-to-beat left ventricular end-diastolic volume (EDV), cardiac output (CO), and maximum left ventricular elastance (E (max)) may also be monitored. To evaluate the technique, we measured aortic pressure waveforms, reference EF and EDV via standard echocardiography, and other cardiovascular variables from six dogs during various pharmacological influences and total intravascular volume changes. Our results showed overall EF and calibrated EDV root-mean-squared-errors of 5.6% and 4.1 mL, and reliable estimation of relative E (max) and beat-to-beat CO changes. These results demonstrate, perhaps for the first time, the feasibility of estimating EF from only a blood pressure waveform. PMID- 19308733 TI - Surface-bound proteins with preserved functionality. AB - Biocompatibility of materials strongly depends on their surface properties. Therefore, surface derivatization in a controllable manner provides means for achieving interfaces essential for a broad range of chemical, biological, and medical applications. Bioactive interfaces, while manifesting the activity for which they are designed, should suppress all nonspecific interaction between the supporting substrates and the surrounding media. This article describes a procedure for chemical derivatization of glass and silicon surfaces with polyethylene glycol (PEG) layers covalently functionalized with proteins. While the proteins introduce the functionality to the surfaces, the PEGs provide resistance against nonspecific interactions. For formation of aldehyde functionalized surfaces, we coated the substrates with acetals (i.e., protected aldehydes). To avoid deterioration of the surfaces, we did not use strong mineral acids for the deprotection of the aldehydes. Instead, we used a relatively weak Lewis acid for conversion of the acetals into aldehydes. Introduction of alpha,omega-bifunctional polymers into the PEG layers, bound to the aldehydes, allowed us to covalently attach green fluorescent protein and bovine carbonic anhydrase to the surfaces. Spectroscopic studies indicated that the surface-bound proteins preserve their functionalities. The surface concentrations of the proteins, however, did not manifest linear proportionality to the molar fractions of the bifunctional PEGs used for the coatings. This finding suggests that surface-loading ratios cannot be directly predicted from the compositions of the solutions of competing reagents used for chemical derivatization. PMID- 19308734 TI - Religious conversion in a psychotic individual. AB - The authors describe the case of a man who appeared to have psychotic symptoms, including self-injurious behavior, but who understood his own experience as a religious conversion. The symptoms, clinical course, and treatment response are described with reference to the works of Kurt Schneider and William James. Empirical studies of the attitudes of psychiatrists, psychiatric patients, and clergypersons about the relationship between religious belief and psychiatric illness are described, and various theoretical models used to understand this relationship are articulated. PMID- 19308735 TI - The involvement of mitochondria and the caspase-9 activation pathway in rituximab induced apoptosis in FL cells. AB - Despite the wide use of anti-CD20 antibody rituximab in the cancer treatment of B cell malignancies, the signalling pathways of CD20-induced apoptosis are still not understood. By using dominant negative (DN)-caspase-9 overexpressing follicular lymphoma cells we demonstrated that the activation of caspase-9 was essential for rituximab-mediated apoptosis. The death receptor pathway mediated by caspase-8 activation was not involved in rituximab-mediated apoptosis since overexpression of FLIP(short) or FLIP(long) proteins, inhibitors of caspase-8 activation, could not inhibit rituximab-induced apoptosis. However, the death receptor pathway activation by anti-Fas antibodies showed an additive effect on rituximab-induced apoptosis. The stabilisation of the mitochondrial outer membrane by Bcl-x(L) overexpression inhibited cell death, showing the important role of mitochondria in rituximab-induced apoptosis. Interestingly, the rituximab induced release of cytochrome c and collapse of mitochondrial membrane potential were regulated by caspase-9. We suggest that caspase-9 and downstream caspases may feed back to mitochondria to amplify mitochondrial disruption during intrinsic apoptosis. PMID- 19308736 TI - The role of visual and phonological representations in the processing of written words by readers with diagnosed dyslexia: evidence from a working memory task. AB - The aim of the study was to elucidate the nature and efficiency of the strategies that readers with phonological dyslexia use for temporary retention of written words in Working Memory (WM). Data was gathered through a paradigm whereby participants had to identify serially presented written (target) words from within larger word pools according to their presentation order, with word pools containing code-specific distracter (CSD) words and non-code-specific distracter (NCSD) words. Analyses focused on three aspects of performance: (1) false recognition of target words; (2) correct recognition of target words; and (3) retention of word presentation order. Participants were readers with diagnosed phonological dyslexia (n = 20, mean grade level = 9.05 [0.89]) and a control group of regular readers (n = 25, mean grade level = 9.00 [0.76]). Results provide direct evidence that the dyslexic readers and the regular readers used essentially different memory coding strategies for the temporary retention of written words, with the former predominantly relying on a visual strategy and the latter on a phonological strategy. Findings further pinpointed a notably impoverished ability of the dyslexic readers to retain word presentation order. The implication of these findings is discussed in relation to theories predicting the acquisition and mastery of reading. PMID- 19308737 TI - Treatment of low-grade gastric MALT-lymphoma unresponsive to Helicobacter pylori therapy: a pooled-data analysis. AB - The most favourable therapeutic strategy for gastric MALT-lymphoma not responding to Helicobacter pylori eradication still remains unclear, neither official guidelines nor randomised studies being available. We therefore performed a systematic review of the literature to evaluate the efficacy of different therapeutic approaches in these patients. Data regarding 315 patients were valuable, and lymphoma remission following the first therapeutic attempt was achieved in 90.1% cases. The most used therapy was radiotherapy (112 patients), followed by surgery (80 patients) and chemotherapy (68 patients), whilst a combination therapy was less frequent. Radiotherapy achieved a higher remission rate as compared to chemotherapy (97.3 vs. 85.3%; P = 0.007), being similar to surgery (97.3 vs. 92.5%; P = 0.2). No difference emerged when comparing lymphoma remission rate achieved by a single therapy with that of combined treatments (89.6 vs. 96.4%; P = 0.6). This is the first pooled-data analysis assessing the efficacy of different oncologic therapeutic approaches to treat gastric MALT lymphoma unresponsive to H. pylori eradication. Radiotherapy seems to be the most suitable treatment in these patients. PMID- 19308738 TI - Altered iron metabolism, inflammation, transferrin receptors, and ferritin expression in non-small-cell lung cancer. AB - The involvement of iron and inflammation parameters on overall survival in non small-cell lung cancer (NSCLC) patients was studied. Furthermore, transferrin receptors 1 (TfR1) and ferritin expression in tumor tissue, tumor stroma, and normal lung tissue were analyzed. Iron metabolism and inflammation parameters were determined by automated laboratory measurements at the time of diagnosis. TfR1 and ferritin expression were determined by immuno-histochemical methods. About 50% of patients survived 12 months only. At the time of diagnosis more than half of the patients had anemia and significantly elevated serum ferritin. Iron content of serum ferritin (ICF) was below the reference values in 90% of patients. Furthermore, ICF showed positive correlation with iron metabolic parameters and survival but negative correlation with serum ferritin and ESR. The expression of TfR1 and ferritin in tumor cells was observed in 88% or 62% of patients, respectively. Tumor stroma was TfR1 negative and sporadically ferritin positive. Tumor tissue ferritin expression showed negative correlation with serum iron and hematokrit (Ht), and positive correlation with ferritin, erythrocyte sedimentation rate (ESR), alpha-1 globulin, and alpha-2 globulin. Positive correlation was found between TfR1 expression in tumor tissue and alpha-globulin. The correlation between TfR1/ferritin expression in tumor tissue and ICF or survival was not observed. Therefore, we conclude that elevated serum ferritin in sera of NSCLC patients is the result of inflammation and oxidative stress rather than body iron overload. Higher expression of ferritin in tumor tissue may be the consequence of iron deficiency or local toxicity induced by environmental factors. PMID- 19308739 TI - Risk stratification for recurrence in patients with esophageal and junctional carcinoma treated with neoadjuvant chemotherapy and surgery. AB - AIM: To stratify the risk for recurrence in patients with esophageal cancer treated with neoadjuvant chemotherapy and surgery. MATERIALS AND METHODS: The prognostic and predictive factors were analyzed in 62 patients who underwent curative resection following chemotherapy. The factors found significant on multivariate analysis were stratified into good, intermediate and high risk groups for recurrence. RESULTS: Kaplan-Meier survival at 3 and 5 years was 32% and 20%, respectively, with a median survival of 19 months. Pathological response and percent node positive were the significant factors on multivariate analysis. Three groups were formed and their recurrence free survivals were calculated using Kaplan-Meier method. The low risk composed of good responders and patients with less than 20% positive lymph node; the intermediate risk composed of non responders and patients with less than 20% positive lymph node and the high risk group composed of non-responders and patients with more than 20% positive lymph node. The median recurrence time was 8 months for the high risk group, 39 months for the intermediate group, and it has not reached in the low risk group. Hazard ratio was 0.39(95% C.I. 0.09-0.98) for the risk group low to intermediate, 0.1(95% C.I. 0.04-0.25) for the low to high risk group and 0.26(95% C.I. 0.11 0.66) for the intermediate to high risk group. CONCLUSIONS: Pathological response rate and percent node positive were significant predictive factors on multivariate analysis. Stratification based on these two predictive factors may help in optimizing any adjuvant treatment. PMID- 19308740 TI - Management in the emergency room of patients requiring hospital treatment of community-acquired pneumonia. AB - INTRODUCTION: To identify factors influencing decisions in initial management of community-acquired pneumonia (CAP) admitted to hospital through Emergency departments. METHODS: Records of CAP adult patients admitted to 24 Spanish hospitals in January-March 2003 were reviewed. Patients sent for ambulatory treatment were excluded. RESULTS: 341 patients (67.0 +/- 24.6 years; 65.3% males) were included; 39% were taking antibiotics at attendance. PSI was (% patients): I II (19.7%), III (14.7%), and IV-V (65.6%). Comorbidities were: COPD (37.2%), heart disease (24.6%), hypertension (17%), diabetes mellitus (10.8%), and malignancies (10%). Pneumococcal/Legionella urinary antigens were performed in 34.0%/42.2% patients. Fewer (p < or = 0.006) rapid tests were performed in class IV-V (p = 0.001), with higher (p < or = 0.01) pneumococcal positive results in class V. Initial treatment was fluoroquinolone (37.5%), beta-lactam + macrolide (26.4%), beta-lactam (22.9%), macrolide (4.7%), and others (8.5%). Patients referred to Internal Medicine had higher heart disease (p = 0.06) and hypertension (p = 0.001) as comorbidity than those at Short-Stay Units or Pneumology. COPD patients were equally distributed between Internal Medicine and Pneumology, with differences vs. Short-Stay Units. CONCLUSIONS: Rapid diagnostic tests were underused, maybe due to broad empirical treatments covering drug resistant pneumococci and L. pneumophila (regardless PSI and comorbidity). Presence of comorbidities or positive results in rapid diagnostic tests seems to influence the medical ward to which the patient is referred to, but not initial treatment. PMID- 19308741 TI - [Multicenter national survey on infection management in patients with penicillin allergy]. AB - INTRODUCTION: Beta-lactam antibiotics are widely prescribed to treat many infections because of efficacy, spectrum and safety. Their use is limited in patients with resistant microbial agents and in those with a history of penicillin allergy (HPA) because of cross-reactivity risk. Accurate clinical assessment of possible HPA requires specialized resources not always available in clinical practice. We intended to get to know the opinion of Spanish physicians about frequency and methods of evaluation of the patients with HPA as well as the preferences in the use of antimicrobial alternatives for common infectious diseases in patients with HPA. METHODS: Multicentric cross-sectional descriptive study ran by the Infectious Diseases Study Group of the Spanish Society for Internal Medicine based on the accomplishment of a survey of 10 questions of opinion to specialist doctors who work in Spanish medical centers. RESULTS: A total of 311 doctors responded to the survey (92.2% Internal Medicine specialists) distributed by all the Spanish territory. An average of 10.7% of patients self-reported having HPA although only an average of 10.8% of them gave documentation on the matter. Patients were sent for specific allergy tests in an average of 33.6%. Desensitization treatments were performed in 4.3% of cases. The preferences for alternative antibiotic therapy to beta-lactam widely varied according to the infectious picture and the communitarian or nosocomial origin, and they included quinolones, macrolides, glycopeptides, lincosamides, oxazolidinones and tigecycline. CONCLUSIONS: Perception about frequency and evaluation of patients with HPA is very variable, but in more than half the cases it is above 10% of the patients. Yet, only one third are sent for in-depth study for allergy confirmation, and less than 5% are desensitized. Nosocomial infections and the possibility of multiresistant bacteria make substantially difficult the management of patients with HPA. PMID- 19308742 TI - [Compliance rate of antibiotic therapy in patients with acute pharyngitis is very low, mainly when thrice-daily antibiotics are given]. AB - OBJECTIVE: To assess drug-compliance observed among patients with suspected streptococcal pharyngitis treated with twice-daily antibiotic regimens (b.i.d.) and others with thrice-daily regimens (t.i.d.). METHODS: A prospective study in the primary care setting was designed in which patients with pharyngitis and three or more Centor criteria, non-allergic to beta-lactam agents, treated with several b.i.d and t.i.d antibiotic regimens based on doctor's choice, were recruited. Patient compliance was assessed with electronic monitoring. RESULTS: A total of 113 patients were enrolled (64 in the t.i.d. group and 49 in the b.i.d. group). Mean openings ranged from 70.3 to 83.3% of the total amount of pills. All the parameters analysed indicated significantly worse compliance with the t.i.d. regimens. Eleven patients allocated to the t.i.d. group took at least 80% of the pills (17.2%), this being significantly lower than those who received b.i.d. antibiotics (59.2%; p < 0.001). The percentage of patients who opened the Medication Event Monitoring System (MEMS) container the satisfactory number of times a day was systematically lower among t.i.d. regimens, this being statistically significant from day three (p < 0.05). Patients assigned to t.i.d. regimens more frequently forgot the afternoon dose. CONCLUSION: Compliance rate was very low, mainly when patients are given t.i.d antibiotic regimens. This may lead to storage of antibiotics and subsequent self-medication. New strategies addressed to improve drug-compliance with antibiotics among outpatients are therefore necessary. PMID- 19308743 TI - [In vitro activity of fosfomycin against ESBL-producing enterobacteria of urinary origin]. AB - In vitro activity of fosfomycin, compared with other antibiotics used for urinary tract infections (UTI), against extended spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae clinical isolates obtained from UTIs, was determined. The activity of fosfomycin, co-trimoxazole, ciprofloxacin, nitrofurantoin, amoxicillin/clavulanic acid and gentamicin against 71 ESBL producing E. coli clinical isolates and 13 ESBL-producing K. pneumoniae clinical isolates obtained from UTI was studied by the agar-dilution method or E-test. E. coli isolates produced mainly CTX-M type ESBL (76.1%), especially CTX-M 14 (56.3%). K. pneumoniae isolates produced most predominantly SHV-type ESBL (92.3%), mainly SHV-2 (76.9%). Gentamicin (4.4%), fosfomycin (5.6%) and nitrofurantoin (5.6%) showed the lowest resistance proportions against E. coli. Co-trimoxazole and ciprofloxacin (7.7%) showed the lowest resistance proportions against K. pneumoniae. PMID- 19308744 TI - [Resistant mutant prevention concentration of fluoroquinolones in clinical isolates of extended spectrum beta-lactamase (ESBL) producing and non-producing strains of Escherichia coli]. AB - The aim of this job is to study the capacity of ciprofloxacin and levofloxacin in restricting the development of resistant mutants in strains of Escherichia coli by determining the mutant prevention concentration (MPC). Ninety-nine isolates of E. coli with different fluoroquinolone susceptibilities were studied and divided into ESBL-producing (n = 60) and non-ESBL producing (n = 36) groups. MPC values were determined using an inoculum of 1010 cfu/ml on Mueller-Hinton plates with serial dilutions of the antibiotics. No significative differences were detected in MIC of ESBL-producing and non-ESBL producing strains of E. coli. Ciprofloxacin susceptible ESBL-producing strains exhibit higher MPC for ciprofloxacin and levofloxacin than non-ESBL producing strains. Our study helps to explain the frequent fluoroquinolone resistance found in ESBL-producing strains. In this context, doubts emerge about the advisability of using fluoroquinolones to treat infections caused by ESBL-producing strains. PMID- 19308745 TI - Viral Load in Spanish HIV patients: trends since the introduction of HAART. AB - The aim of this study is to describe trends in the percentage of samples with undetectable HIV viral load in Spain after the implementation of HAART. A descriptive observational study of HIV-VL measurements carried out in the microbiology department of the Hospital Clinico Universitario de Valladolid (HCUV) was conducted over a 9-year period (1996-2004). Regarding the trend over the study period, the 30-39 years age group accounted for most of the samples, although the percentage decreased from 65.5% to 59.6% over the study period. In contrast, the 40-49 years group increased from 9.1% to 14.5%. The preponderance of men, with percentages above 70%, was observed during the whole period. Although the purpose of this treatment is to maintain undetectable viral loads, since 1999 more than 60% of nonfirst samples had detectable levels. Based on the results of the VL trend among HIV/AIDS patients observed in this study, a large number of patients maintain elevated detectable VL years after HAART was implemented. Although different factors may be the cause of this and should be delimited in future studies, the phenomenon observed demonstrates the usefulness of monitoring VL and analyzing its time trend to gain further knowledge about the therapeutic results and care of HIV patients as a whole, also serving as the basis for corrective measures. PMID- 19308746 TI - [Importance of the antimicrobial spectrum and the bacterial resistances in the antibiotic choice for the treatment of pediatric patients with communitary infections]. AB - OBJECTIVE: This study aimed to know the importance of the antimicrobial spectrum and the bacterial resistances for the antibiotic choice in the extrahospitalary pediatric area, at the same time that establish the relationship with others therapeutics parameters. MATERIAL AND METHODS: Cross-sectional, observational study within the MUSA (Improvement of Use of Antimicrobial Agents in Primary Health Care) Project made by personal interview of 210 pediatrician doctors randomly selected with national representation. This target is included in a bigger universe (855 doctors) representative of the most doctors responsible of the antibiotic prescriptions in the extrahospitalary area (sample error of the 3.3% for a 95% confidence interval and maximum response dispersion: p=q=50). The results of the pediatric study were subjected to a comparative analysis with the results of a similar study made ten years ago and with the global results of the general study. The questionnaire used for the interview had two clearly different parts: in the first part, the questions were open with the objective to get spontaneous answers from the participants; the second part had questions with suggested answers. RESULTS: Clinical efficacy is the most spontaneously valued issue by the Spanish pediatricians when it comes to choosing an antimicrobial agent; efficacy is followed by tolerance/safety and posology. Antimicrobial spectrum is mentioned by one out of 4-5 pediatricians that have participated in the study (21.9%), while the bacterial resistances are only mentioned by a 3.8%. In a suggested level, clinical efficacy is still the most valued parameter, being identified by 7 out of 10 interviewed with the bacterial eradication. In this case, the antimicrobial spectrum is the following parameter on significance, appearing as a synonym of "activity against specific microorganisms" in 2 out of 3 cases. In relation to his own evaluation, 3 out of 4 pediatricians say they take "a lot" of the bacterial resistances into consideration, who are mostly understood as "global rate" for 5-6 out of 10 interviewed, whether the resistance showed by the pneumococcal is what worried the most to 4 out of 10 of them. This importance given to the bacterial resistances at the second part of the study contrasts with the low consideration showed when it is compared with other parameters and the valoration is treated in a spontaneous way. In relation to a similar study realized on 1997 (Urano Proyect), we observe that clinical efficacy has replaced tolerance/safety as a parameter to bear in mind, probably as a consequence of more experience and confidence with the most important antimicrobial agents in the antibiotic prescriptions in podiatry. On the other hand, the bacterial resistances still being left out on the pediatricians spontaneous consideration, a fact that is shared with the majority of the collectives participating on the general study. The rest of the parameters stay in a similar way. CONCLUSIONS: Is necessary to emphasize at the importance of bacterial resistances in the antimicrobials rational use on the pediatric patient. As the antimicrobial tolerance profile has been improving in the last ten years, clinical efficacy, understood as "bacterial eradication" has became the most determining parameter when choosing antimicrobial agents. PMID- 19308747 TI - [Clinical experience with tigecycline in the treatment of nosocomial infections caused by isolates exhibiting prevalent resistance mechanisms]. AB - This article reviews the clinical experience with tigecycline in the treatment of infections caused by microorganisms with prevalent resistance mechanisms among nosocomial microbiota, as methicillin-resistant Staphylococcus aureus, vancomycin resistant enterococci, multidrug- resistant Acinetobacter baumannii and enterobacteria producing extended spectrum beta-lactamases. Most of articles found in the literature describe the use of tigecycline in the treatment of severe infections (sepsis and septic shock, nosocomial pneumonia and ventilator associated pneumonia...) produced by multidrug-resistant microorganisms, in patients with multiple comorbidities (admitted in ICU, with malignancies, transplants and/or immunodepressed...) and in many occasions after failures of previous antibiotic treatments. Favourable outcomes with tigecycline are reported in most articles. However, an accurate global assessment is difficult since, in addition to the described confounding factors, there are concomitant or sequential antibiotic treatments in several communications, and lack of relevant clinical (as comorbidities), microbiological (as susceptibility) and outcome (different criteria by different authors) data in others. More even, the described series are retrospective and lack of control groups. Nevertheless the usefulness of this revision is based on the fact that in daily clinical practice the use of tigecycline will increase, since epidemiology of specific hospital medical units shows multidrug resistance among nosocomial isolates and tigecycline can be one of the scarce available compounds active against multidrug resistant strains/clones. PMID- 19308748 TI - Angiotensin II type 1 and 2 receptors gene polymorphisms in pre-eclampsia and normal pregnancy in three different populations. AB - OBJECTIVES: To investigate a possible association between pre-eclampsia (PE) and the genotype for the angiotensin II type-1 receptor (AT1R) and the angiotensin type-2 receptor (AT2R) in various population groups. DESIGN: The study was retrospective in a case-controlled design. SAMPLES: Two hundred thirty-six pregnant women with PE/eclampsia (E) and 426 non-hypertensive pregnant women were included. METHOD: Polymorphic sites of AT1R (A1166C) and AT2R (A1675G) were amplified by polymerase chain reaction, digested with a restriction enzyme that differentiated between the alternative alleles, and analyzed. MAIN OUTCOME MEASURES: Maternal genotypes and their correlation with clinical parameters. RESULTS: The frequency of the AT2R-GG genotype (A1675G) in the PE group was significantly greater than in controls for Afro-Caribbean women (49.3% vs 26.9%, p=0.004), but the frequency difference in Asian or Caucasian women was not significant (23.0% vs 25.4%, p=0.63; 27.7% vs 14.8%, p=0.17, respectively). The highly significant difference in Afro-Caribbean women was maintained after controlling for the effects of age, BMI and parity (p=0.005). There was no significant association of the molecular variant of AT1R (A1166C) with PE in Afro Caribbean, Caucasian or Asian women. However, in the whole PE group compared to the controls there was a higher proportion of the AT2R-GG genotype with AT1R-AC (56% vs 44%, OR 2.37; 95% CI: 1.06-5.32). In Afro-Caribbean women, the combination of AT1R-AC with AT2R-AG genotypes was significantly higher in controls compared to PE group (93.8% vs 6.3%, OR 0.11; 95% CI: 0.01-0.81). CONCLUSION: There is an association between PE/E and the GG-genotype of AT2R in Afro-Caribbean women. PMID- 19308749 TI - Validation of epithelial ovarian cancer and fallopian tube cancer and ovarian borderline tumor data in the Danish Gynecological Cancer Database. AB - OBJECTIVE: To validate the data on epithelial ovarian cancer, fallopian tube cancer and borderline ovarian tumors registered in the nationwide Danish Gynecological Cancer Database (DGCD) in 2005 and 2006. The DGCD is a multidisciplinary database that contains data for research and quality improvement. DESIGN: Comparative registry-based study supplemented with data from medical records. SETTING: Six hospitals in Denmark. PARTICIPANTS: Women registered with epithelial ovarian cancer, fallopian tube cancer and borderline ovarian tumor. MAIN OUTCOME MEASURE: Data completeness and strength of agreement. RESULTS: The estimated completeness of reporting to the DGCD was 94.2% and the strength of agreement between the variables in the DGCD and the medical file varied from moderate to very good. The important quality indicator 'complication' had the lowest strength of agreement. CONCLUSION: The validity of ovarian cancer data in the DGCD is sufficient for quality monitoring in gynecological oncology. PMID- 19308750 TI - Oxidative stress but not endothelial dysfunction exists in non-obese, young group of patients with polycystic ovary syndrome. AB - OBJECTIVE: To determine if oxidative stress and endothelial dysfunction exist at the same time in a young, non-obese group of patients with polycystic ovary syndrome (PCOS). DESIGN: Cross-sectional study. SETTING: Celal Bayar University, School of Medicine, Manisa, Turkey. SAMPLE: Thirty-one young, non-obese patients with PCOS and 23 age- and body mass index-matched controls. METHODS: Following clinical and biochemical diagnosis, malonyldialdehyde (MDA), superoxide dismutase (SOD), von Willebrand Factor (vWF), and nitric oxide (NO) levels of patients and controls were measured and compared. MAIN OUTCOME MEASURES: To find out oxidative stress and endothelial dysfunction parameters. RESULTS: MDA (0.12+/-0.03 vs 0.10+/-0.03, p=0.01) and SOD (8.0+/-0.7 vs 7.28+/-0.8, p=0.001) levels were significantly higher in PCOS group while there was no difference in vWF (527.2+/ 280.1 vs 568.1+/-276.8, p>0.05) and NO levels (169.9+/-47.4 vs 168.9+/-80, p>0.05). When the results of the PCOS patients were divided into two subgroups in terms of insulin resistance (IR- and IR + ), the IR- subgroup had significantly higher MDA levels compared to the control (0.125+/-0.03 vs 0.101+/-0.03, p=0.03). Though IR+ group also had higher MDA than the control group, it did not reach to a significant level (0.117+/-0.05 vs 0.101+/-0.03, p>0.05). Both IR- and IR+ groups had significantly higher SOD levels compared with control group (7.99+/ 0.7 vs 7.28+/-0.8, p=0.009 and 8.22+/-0.8 vs 7.28+/-0.8, p=0.03, respectively). vWF and NO levels were not different among these three groups (p>0.05). CONCLUSIONS: Oxidative stress is prominent while endothelial dysfunction does not exist in young, non-obese patients with PCOS. PMID- 19308751 TI - Maternal serum endostatin at gestational weeks 16-20 is elevated in subsequent pre-eclampsia but not in intrauterine growth retardation. AB - OBJECTIVE: Endostatin, an important anti-angiogenic factor produced by endothelial cells, is elevated in established pre-eclampsia. We measured maternal serum endostatin concentrations in early pregnancy associated with later pre eclampsia and intrauterine growth retardation (IUGR). DESIGN: Retrospective case control study. SETTING: University Central Hospital. SAMPLE: Serum samples were collected at 12-15 and 16-20 gestational weeks from a total of 124 pregnant women of whom 49 developed pre-eclampsia, 16 gave birth to infants with IUGR without pre-eclampsia, and 59 remained normotensive giving birth to healthy, normal weight infants. METHODS: Enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Endostatin concentrations in serum. RESULTS: At 12-15 gestational weeks, there was no difference in median endostatin concentrations between the groups. At 16-20 gestational weeks, the median endostatin concentration was higher in the women with subsequent pre-eclampsia (p=0.026), especially preceding a later severe form of the disease (p=0.041), than in the controls. The results were further confirmed by receiver operating characteristic (ROC) analysis showing an area under the curve (AUC) of 0.64 (95% confidence interval: 0.50 0.81) for endostatin to identify subsequent pre-eclampsia, and 0.71 (0.53-0.89) in cases of severe pre-eclampsia. Optimal cut-off values were determined and used for calculations of sensitivity and specificity, which were 80 and 52% (cut-off value = 58.0 microg/L) in pre-eclampsia, and 80 and 65% (cut-off value = 65.5 microg/L) in the severe form of the disease. CONCLUSIONS: The concentrations of endostatin in maternal serum at 16-20 weeks' of gestation are associated with an increased risk of pre-eclampsia but not IUGR. PMID- 19308752 TI - Smoking worsens the prognosis of mild abnormalities in cervical cytology. AB - OBJECTIVE: To examine the effect of smoking on the incidence of low- and high grade cervical intraepithelial neoplasia (CIN) in women with a baseline Pap smear of atypical squamous cells (ASC) or a low-grade squamous intraepithelial lesion (LSIL). DESIGN: Prospective study in which a cohort of women with normal colposcopy and ASC/LSIL at baseline were followed at 6-month intervals of up to 36 months. Women were grouped in post-hoc analysis according to their smoking behavior: never (or past) smokers and current smokers. SETTING: This report was based on data from the Latin American Screening Study, conducted in Sao Paulo, Campinas, Porto Alegre (Brazil) and Buenos Aires (Argentina). POPULATION: A subset of 150 women derived from a cohort of 1,011 women. METHODS: Multivariate Cox analysis and Kaplan-Meier curves were used. MAIN OUTCOME MEASURES: Low- and high-grade CIN during follow-up. RESULTS: The only factor related to an increased risk of developing CIN was the positive high-risk (hr) HPV status (hazard ratio (HR) = 3.42; 95% CI: 1.11-9.43). A total of 21 cases of incident CIN were detected during follow-up. Of these, 11 appeared in the group of 67 smokers and 10 among the 83 non-smoker women (log-rank, p=0.33). Smoking status was not associated with the risk of developing CIN (HR = 0.73; 95% CI: 0.40-1.33). However, when restricting the analysis to high-grade CIN only (11 cases), the probability of developing the disease was significantly higher among smokers (p=0.04). CONCLUSIONS: Smoking contributes additional risk for developing high grade CIN in women with ASC or LSIL cytology but normal colposcopy. PMID- 19308753 TI - Concentrations of vascular endothelial growth factor C and D in amniotic fluid and maternal plasma. AB - OBJECTIVE: Vascular endothelial growth factor-C (VEGF-C) and VEGF-D promote both endothelial and lymphatic vascularization during embryonic development. We studied their presence in amniotic fluid (AF) and maternal plasma during pregnancy. DESIGN: Descriptive study. Setting. University Central Hospital, Helsinki, Finland. Samples. AF at 14-20 weeks (n=20) and 38-41 weeks' of gestation (n=20), serial plasma samples (n=38) from 16 healthy pregnant women during 8-40 weeks of pregnancy, and plasma from 15 non-pregnant controls. METHODS: Enzyme-linked immunosorbent assay (ELISA). Main outcome measures. Concentrations of VEGF-C and -D in AF and maternal plasma. RESULTS: VEGF-C concentrations in AF decrease as pregnancy advances (p=0.002) and are about 10 fold lower than in plasma at the corresponding gestational age (p=0.001). Plasma VEGF-C concentrations are higher in the first trimester (p=0.014) and the early second trimester (p=0.016) than in non-pregnant women. VEGF-D is not detectable in AF, but in plasma its concentrations become higher at term compared with non pregnant women (p=0.039). CONCLUSIONS: VEGF-C and -D are present in high concentrations in maternal plasma, while only VEGF-C can be detected in AF. PMID- 19308754 TI - Use of statistical methods in dental research: comparison of four dental journals during a 10-year period. AB - OBJECTIVE: To investigate development of the use of statistical methods in dental journals in 1996, 2001, and 2006 hypothesizing that methods are becoming more sophisticated. MATERIAL AND METHODS: All original research articles in four dental journals in 1996, 2001, and 2006 were reviewed: Journal of Dental Research (JDR), Caries Research (CR), Community Dentistry and Oral Epidemiology (CDOE) and Acta Odontologica Scandinavica (AOS). Evaluation covered 928 articles reporting original research findings based on systematic collection and statistical analysis of research data. Aspects measured were: research design, statistical methodology, description of procedures, and presentation of results. Percentage frequencies of reported statistical procedures were compared between journals and between years. RESULTS: The main findings of the study are: 1) that use of multivariate or specific methods did not increase from 1996 to 2006, 2) that traditional statistical significance testing is still widely used in dental journals and that use of confidence intervals increased in the period 1996 to 2006, 3) that statistical procedures are usually extensively described, and 4) that dental journals have different profiles in their statistical content. CONCLUSIONS: The authors of articles in dental journals might apply these results when designing their dental research, i.e. to use statistical methods and to present results in line with the policy and presentation of the leading dental journals. PMID- 19308755 TI - Accelerated partial breast cancer irradiation (APBI)--the future breast cancer radiotherapy? PMID- 19308756 TI - Oxidative stress in non-small cell lung cancer: role of nicotinamide adenine dinucleotide phosphate oxidase and glutathione. AB - BACKGROUND: Cigarette smoke is strongly associated with NSCLC, but the carcinogenesis of NSCLC is poorly understood. METHODS: To discover the role of oxidative stress and anti-oxidative defense in NSCLC, we measured NADPH oxidase (NOX) activity, myeloperoxidase activity, 8-OHdG, and glutathione content from lung specimens. These came from 32 patients: 22 NSCLC patients and ten controls without cancer. RESULTS: In NSCLC patients, NOX activity was significantly higher both in the malignant (p = 0.001) and non-malignant (p = 0.044) samples from NSCLC patients, than in the control specimens. Myeloperoxidase activity was lower (p = 0.001) and glutathione content (p = 0.009) higher in malignant tissue. No significant difference was observable in 8-OHdG content between patient groups. CONCLUSIONS: Increase in NOX activity in the malignant tissues was independent of smoking history and myeloperoxidase activity, suggesting its independent role in NSCLC pathogenesis. PMID- 19308757 TI - Persistent neuropathy after treatment with cisplatin and oxaliplatin. AB - BACKGROUND: The aims of the current explorative study were to assess persistent neuropathy in 45 patients up to 6 years after treatment with cisplatin or oxaliplatin and to determine the most adequate method to evaluate neuropathy. Furthermore, the effect of possible determinants on persistent neuropathy was investigated. MATERIAL AND METHODS: The assessment of neuropathy was performed using a questionnaire, by neurological tests, and by vibration threshold (VT) measurements. Because VT determination gives the most objective information, VT measurements were used for further analyses. RESULTS AND DISCUSSION: The analyses revealed that neuropathy of the hands was related to follow-up time, with an observed recovery half-life of 6.8 (+/- 3.1) years. No significant reversibility of neuropathy of the feet within the observation period could be demonstrated. For cisplatin, the severity of neuropathy was related to the cumulative dose and sodium thiosulfate use. Oxaliplatin induced neuropathy did not appear to be related to the dose within the studied dose range. No relationship with platinum levels, renal function, glutathione transferase genotypes, diabetes mellitus, alcohol use, or co-medication could be demonstrated. This study was performed as an explorative study and the issues discussed need to be investigated further. PMID- 19308758 TI - Unusual sequence of lymphoid disorders: follicular lymphoma subsequent to Hodgkin lymphoma and transformed into diffuse large B-cells non Hodgkin lymphoma. PMID- 19308762 TI - Optimization of computed tomography urography protocol, 1997 to 2008: effects on radiation dose. AB - BACKGROUND: Since computed tomography (CT) urography began to replace excretory urography as the primary imaging technique in uroradiology, the collective radiation dose to the patients has increased. PURPOSE: To examine the changes in the CT urography protocol for investigating suspected urinary tract malignancy between the years 1997 and 2008, and how these changes have influenced the mean effective dose. MATERIAL AND METHODS: The study was based on 102 patients (mean age 66.1+/-14.8 years, range 31-89 years; 30 female, 72 male) divided into five groups (groups A-E) corresponding to the time points at which changes were made to the CT urography protocol. The mean effective doses were estimated using the ImPACT CT Patient Dosimetry Calculator. RESULTS: The number of scan phases at CT urography was reduced from four to three in 1999, resulting in a reduction of the mean effective dose from 29.9/22.5 (female [F]/male [M]) mSv (group A) to 26.1/18.9 (F/M) mSv (group B). In 2001, mAs settings were adapted to patient size, and the mean effective dose was reduced to 16.8/12.0 (F/M) mSv (group C). In 2005, scans were performed with a multidetector-row CT equipped with automatic tube current modulation in the x- and y-axis (CARE Dose). The effective mAs was also lowered in the unenhanced and excretory phase, yet the mean effective dose increased to 18.2/13.1 (F/M) mSv (group D), since the effective mAs had to be increased in the corticomedullary phase to maintain image quality. In 2008, as tube current modulation in the x-, y-, and z-axis was introduced (CARE Dose4D), the mean effective dose was reduced to 11.7/8.8 (F/M) mSv (group E). CONCLUSION: This study shows that the individual mean effective dose to patients undergoing CT urography has decreased by 60%, from 29.9/22.5 (F/M) mSv in 1997 to 11.7/8.8 (F/M) mSv in 2008. PMID- 19308763 TI - Magnetic resonance coronary angiography: comparison between a Gd-BOPTA- and a Gd DTPA-enhanced spoiled gradient-echo sequence and a non-contrast-enhanced steady state free-precession sequence. AB - BACKGROUND: Several studies have demonstrated that the administration of contrast agents is advantageous in magnetic resonance coronary angiography (MRCA). PURPOSE: To compare a non-contrast-enhanced steady-state free-precession (SSFP) with a contrast-enhanced inversion recovery spoiled gradient-echo (IR-GE) sequence using two different contrast agents for MRCA. MATERIAL AND METHODS: Eight healthy volunteers were examined on a 1.5T MR scanner. For non-contrast enhanced MRCA, a breath-hold three-dimensional (3D) SSFP sequence (repetition/echo time [TR/TE] 3.9/1.7 ms, flip angle [FA] 65 degrees) was used. Contrast-enhanced MRCA was performed repetitively in two imaging sessions over 30 min after injection of 0.2 mmol/kg body weight gadobenate dimeglumine (Gd-BOPTA) or gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA) using a breath-hold 3D IR-GE sequence (TR/TE 4.1/1.7 ms, FA 15 degrees). The signal-to-noise ratios (SNR) of the coronary arteries, as well as the contrast-to-noise ratios (CNR) between coronary arteries and perivascular tissue, were calculated for all images. Blood T (1) values were repetitively estimated over 30 min using an SSFP sequence with incrementally increasing inversion times (TR/TE 2.4/1.0 ms, FA 50 degrees). RESULTS: Gd-BOPTA-enhanced images showed significantly (P<0.05) higher SNR and CNR compared to Gd-DTPA-enhanced images for all times after contrast injection (SNR: 1 min post injection [PI] 26.4+/-4.2 vs. 16.2+/-3.1; CNR: 1 min PI 21.4+/-3.7 vs. 13.2+/-2.6). Compared to the SSFP images, the Gd-BOPTA-enhanced images showed higher CNR values for all times after injection (1 min PI 21.4+/ 3.7 vs. 13.8+/-5.5; P<0.05), whereas the Gd-DTPA-enhanced images did not (1 min PI 13.2+/-2.6 vs. 13.8+/-5.5; P>0.05). Blood T (1) estimates were not significantly different for either agent 1 min after administration (P>0.05), but they were significantly lower for Gd-BOPTA (P<0.05) from 7 to 25 min after injection. CONCLUSION: Compared to non-contrast-enhanced SSFP images, only Gd BOPTA-enhanced images show a significantly improved contrast between the coronary arteries and the surrounding tissue. PMID- 19308764 TI - Solitary fibrous tumor of the lung: three rare cases of intraparenchymal nodules. AB - Solitary fibrous tumor (SFT) of the pleura usually presents as a peripheral mass, in contact with the surface of the pleura. However, on occasion, it can occur separately from the pleura, in the lung parenchyma. We describe the radiological and imaging features of three SFTs of the lung, diagnosed in our department, with relevant clinical data. The diagnosis of SFT of the lung, although rare, should be considered in a slow-growing solitary lung parenchymal nodule. PMID- 19308765 TI - Usage of support services in primary lateral sclerosis. AB - Patients with amyotrophic lateral sclerosis (ALS) rely on a variety of support services during the course of their illness. Patients with primary lateral sclerosis (PLS) have a slower progression of disease and different clinical spectrum. Their needs for allied health services and social support have not been well characterized. To investigate these needs, 25 patients with PLS and caregivers were surveyed on the use of assistive devices and support services. Needs for assistance changed as the disease progressed. Their greatest need was for gait-assistive devices and home help for activities requiring mobility. As in other chronic diseases, there was a striking use of the internet to gather information and for patient support groups. PMID- 19308766 TI - Identifying poor adaptation to a new diagnosis of motor neuron disease: A pilot study into the value of an early patient-led interview. AB - This pilot study explored whether clinicians can identify patients who may not be coping with a diagnosis of MND early in the disease course by using patient-led interviews rather than psychological testing. Consecutive, newly diagnosed MND patients underwent a semi-structured interview six and 18 weeks after diagnosis, and completed SEIQoL-DW, ALSFRS-R, MND Coping Scale, MND Social Withdrawal Scale, SF 36 v2 and HAD. Three physicians independently used a list of factors from the literature associated with coping with a diagnosis of MND, and overall impression to assign patients from interview transcripts to groups of copers and non-copers. Ten of 13 recruited patients were categorized unanimously using the first interview alone. Four patients were categorized as non-copers. These showed significantly higher scores for depression, anxiety and social withdrawal and significantly lower scores for coping and mental composite scores on the SF 36 v2. Our pilot study suggests it is possible for experienced clinicians to identify patients who may struggle to cope with a diagnosis of MND early by patient-led interview alone. This provides some evidence for the therapeutic potential of the early follow-up appointment interview with a senior doctor soon after diagnosis, advocated by current British guidelines. PMID- 19308767 TI - Treatment with lithium carbonate does not improve disease progression in two different strains of SOD1 mutant mice. AB - It has been shown that chronic treatment with lithium carbonate (Li(2)CO(3)) in presymptomatic SOD1G93A transgenic male mice, a model of ALS, was able to remarkably increase their lifespan through the activation of autophagy and the promotion of mitochondriogenesis and neurogenesis. This prompted us to test the lithium effect also in female SOD1G93A mice with two phenotypes of different disease severity. Female SOD1G93A mice of C57BL/6J or 129S2/Sv genetic background were treated daily with Li(2)CO(3) 37 mg/kg (1 mEq/kg) i.p. starting from age 75 days until death. Grip strength, latency to fall on rotarod and body weight were monitored twice weekly. At the time of death the spinal cord was removed to assess the number of motor neurons and to measure the expression of a marker of autophagy (LCII) and the activity of mitochondrial complex IV. We observed a significant anticipation of the onset and reduced survival in 129Sv/G93A and no effect in C57/G93A mice treated with lithium compared to vehicle treated mice. Moreover, lithium neither exerted neuroprotective effects nor increased the expression of LCII and the activity of mitochondrial complex IV in the spinal cord. The present study does not identify any therapeutic or neuroprotective effect of lithium in SOD1G93A female mice. PMID- 19308768 TI - Lessons from an audit of psychoeducation at an older adolescent inpatient unit. AB - OBJECTIVES: The aim of this paper was to determine if patients undergoing treatment at an older adolescent inpatient unit receive psychoeducation according to the unit philosophy of providing timely and pertinent information regarding illness/diagnosis, medication, diet, outpatient follow-up, and alcohol/ drug use. METHODS: Data were gathered prospectively as part of a quality assurance initiative at the Christchurch Youth Inpatient Unit. Patients were interviewed by a registered nurse using a structured audit tool. RESULTS: Participants reported receiving adequate information on medication and illness, and most received information on outpatient follow-up and alcohol and drug use. However, the majority reported a lack of information/ advice about diet. Patients' reported awareness of relapse prevention and the relationship of alcohol and other drugs use to medication and treatment was lower than expected. CONCLUSIONS: The audit highlighted areas of discrepancy between information staff believed they had delivered and information youth perceived as received. Psychiatric staff working with young people need be aware of the timing, language and mode of delivery of psychoeducation to enable their patients to 'take in' the information provided. PMID- 19308769 TI - The role of hypoxia-inducible factor in osteogenesis and chondrogenesis. AB - Oxygen availability is a critical signal for the proper development of many tissues. The transcription factor hypoxia-inducible factor (HIF) plays an important role in maintenance of oxygen homeostasis in all metazoans. Numerous growth and transcription factors have been implicated in the development of the skeletal system. Many of the factors that are up-regulated during hypoxia are downstream of HIF activation. Recent work has demonstrated that HIF has a critical function in cartilage and bone development. We summarize current understanding of the role of HIF in bone and cartilage development. PMID- 19308770 TI - Therapeutic potential of mesenchymal stromal cells in a mouse breast cancer metastasis model. AB - BACKGROUND AIMS: Mesenchymal stromal cells (MSC) have been studied intensively in regenerative medicine. However, their therapeutic potential against tumor formation and cancer metastasis is still unclear. The effects of transplantation of MSCs in early-stage of carcinogenesis, should be evaluated. METHODS: MSC isolated from human umbilical cord blood (UCB) and adipose tissue (AD) were transplanted in a mouse cancer metastasis model. The effects of MSC on tumor growth and metastasis were analyzed. The effects of transplantation of MSC into the mouse model at very early stage carcinogenesis were also evaluated. RESULTS: Human MSC reduced lung metastasis and inhibited the growth of human breast cancer cells by inducing apoptosis. In addition, transplantation of both UCB and AD MSC into a cancer model with no detectable clinical symptoms did not appear to promote tumor growth or metastasis. CONCLUSIONS: We evaluated the effect of MSC derived from human UCB and AD tissue in a tumor model. Our findings may help to elucidate the interaction between cancer cells and MSC, as well as the application of MSC to clinical trials. PMID- 19308771 TI - Clinical-grade ex vivo-expanded human natural killer cells up-regulate activating receptors and death receptor ligands and have enhanced cytolytic activity against tumor cells. AB - BACKGROUND AIMS: Cancer immunotherapy involving natural killer (NK) cell infusions and administration of therapeutic agents modulating the susceptibility of tumors to NK-cell lysis has been proposed recently. We provide a method for expanding highly cytotoxic clinical-grade NK cells in vitro for adoptive transfer following bortezomib treatment in patients with advanced malignancies. METHODS: NK cells were expanded with irradiated Epstein-Barr virus-transformed lymphoblastoid cells. Expanded cells were evaluated for their phenotype, cytotoxicity, cytokine secretion, dependence on interleukin (IL)-2 and ability to retain function after cryopreservation. RESULTS: A pure population of clinical grade NK cells expanded 490+/-260-fold over 21 days. Expanded NK cells had increased TRAIL, FasL and NKG2D expression and significantly higher cytotoxicity against bortezomib-treated tumors compared with resting NK cells. Expanded NK cells, co-cultured with K562 and renal cell carcinoma tumor targets, secreted significantly higher levels of soluble Fas ligand 6; fgjhd IFN-gamma, GM-CSF, TNF alpha, MIP-1alpha and MIP-1beta compared with resting NK cells. Secretion of the above cytokines and NK-cell cytolytic function were IL-2 dose dependent. Cryopreservation of expanded NK cells reduced expression of NKG2D and TRAIL and NK-cell cytotoxicity, although this effect could be reversed by exposure of NK cells to IL-2. CONCLUSIONS: We describe a method for large-scale expansion of NK cells with increased expression of activating receptors and death receptor ligands resulting in superior cytotoxicity against tumor cells. This ex vivo NK cell expansion technique is currently being utilized in a clinical trial evaluating the anti-tumor activity of adoptively infused NK cells in combination with bortezomib. PMID- 19308772 TI - Research on promoting periodontal regeneration with human basic fibroblast growth factor-modified bone marrow mesenchymal stromal cell gene therapy. AB - BACKGROUND AIMS: Recently, it has been found that effective periodontal regeneration can be induced by bone marrow mesenchymal stromal cell (BMSC) transplantation or local application of basic fibroblast growth factor (bFGF). The aim of the present study was to assess, in dogs, the efficacy of periodontal regeneration via the delivery of BMSC transfected with bFGF to repair destruction of periodontal tissue. METHODS: BMSC from dogs were isolated, cultured and purified via density-gradient centrifugation. Polymerase chain reaction (PCR) was employed to clone bFGF cDNA from human periodontal cells, and the product was then ligated into the eukaryotic expression vector pDC316-IREs-EGFP. BMSC transfected with pDC316bFGF-IREs-EGFP were transplanted into root furcation defects of beagle dogs. After 6 weeks, regeneration in defects was assessed via clinical examination, X-ray, histologic observation and micro-CT analysis. RESULTS: DNA sequence analysis showed that the bFGF sequence of recombinant plasmid pDC316bFGF-IREs-EGFP was consistent with that reported by GeneBank. bFGF expression was detected with Western blotting, and active bFGF in supernatant was also observed. Our animal experiment proved that the regenerating speed of periodontal bone tissue in groups transplanted with BMSC containing the modified bFGF gene was higher than in those transplanted with BMSC alone. CONCLUSIONS: A successfully constructed eukaryotic expression vector containing human bFGF in pDC316bFGF-IREs-EGFP could produce bioactive bFGF in vitro. bFGF overexpression mediated by the recombinant plasmid pDC316bFGF- IREs-EGFP accelerated periodontal regeneration. PMID- 19308773 TI - Results of a pilot study on the use of third-party donor mesenchymal stromal cells in cord blood transplantation in adults. AB - BACKGROUND AIMS: Cord blood (CB) transplants with co-infusion of third-party donor (TPD) mobilized hematopoietic stem cells (MHSC) have been shown to result in 'bridge' engraftment with prompt neutrophil recovery and high final rates of CB engraftment and full chimerism. This strategy overcomes the limitation posed by low cellularity of CB units for unrelated transplants in adults. Enhancement of adaptive immunity reconstitution without increasing risks of graft-versus-host disease (GvHD) is required to optimize results further. Our objectives were to evaluate co-infusion of mesenchymal stromal cells (MSC) from the same TPD regarding tolerance, CB engraftment and effects on acute (a)GvHD, both preventive and therapeutic. METHODS: Ex vivo-expanded bone marrow MSC were infused at the time of the transplant or the in case of refractory aGvHD. RESULTS: Nine patients received 1.04 - 2.15 x 10(6)/kg (median 1.20) MSC immediately after CB and TPD MHSC. Neither immediate adverse side-effects nor significant differences regarding CB engraftment or aGvHD development were observed. Four patients developed grade II aGvHD, refractory to steroids in two. These reached complete remission after therapeutic infusions of MSC. CONCLUSIONS: In recipients of 'dual CB/TPD MHSC transplants', MSC infusions were therapeutically effective for severe aGvHD but no significant differences in CB engraftment and incidence of severe aGvHD were observed following their prophylactic use. Although results of this study alone cannot conclusively determine the application of MSC in CB transplantation, we believe that, in this setting, the best use of MSC could be as pre-emptive treatment for aGvHD. PMID- 19308775 TI - Development and validation of an attention-deficit/hyperactivity disorder (ADHD) executive function and behavior rating screening battery. AB - Attention problems are ubiquitous in clinical practice, commonly found in many childhood learning and behavior disorders. Practitioners need cost- and time effective methods for determining whether children have attention problems due to attention-deficit/hyperactivity disorder (ADHD) or numerous other conditions. This study examined the utility of a 15-minute ADHD screening battery designed to differentiate ADHD (including inattentive, IT, and combined, CT, subtypes), specific learning disability (SLD), and typical child samples. Results for the 368 children (age 6 to 12 years) revealed that the Trail Making Test-Part B (Time/Errors), Hale-Denckla Cancellation Test (Time/Correct), and Child Attention Profile (Inattention/Overactivity) teacher ratings discriminated between typical and ADHD groups (87% correct classification; sensitivity = .64; specificity = .92) and differentiated between IT, CT, and SLD groups (80% correct classification; IT sensitivity = .82, and specificity = .96; CT sensitivity = .84, and specificity = .82). Discriminant function and Bonferroni post hoc results revealed different neuropsychological and behavioral patterns among groups. PMID- 19308776 TI - Innovation in medical postgraduate education: a pilot study. AB - Although widely criticized for its didactical inadequacy, the accepted form for medical postgraduate training is still the symposium. One of its great deficiencies is the inability for participants to put new (scientific) findings into a personal medical context by means of discussion and reflection. To meet the didactical demands an innovative approach to postgraduate education was developed and tested with gynaecologist as teachers and general practitioners as participants. The first results with five courses are very positive: in comparison to the symposium, the new approach stimulates internalisation of provided knowledge much more, information retention is much higher and the reactions of participants are overwhelmingly enthusiastic. A short outline of the didactic model and its procedures is provided. PMID- 19308777 TI - Ethnic Turkish fathers in birth support roles in a Berlin labour and delivery room--motives, preparation and incidence in a 10-year comparison. AB - PURPOSE: Have there been changes among German (G) or ethnic Turkish (T) fathers with respect to birth preparation or motivation to participate in a birth support role? In a 10-year comparison is it possible to identify an increase among ethnic Turkish fathers in birth support roles in Germany? METHODS: (1) In 1995/96 and 2003, structured interviews were conducted with German and ethnic Turkish fathers on the second or third post-natal day, focusing on the following topics: birth preparation, motives for participating in the birth, anxieties experienced by the father in the labour and delivery room, the father's assessment of his role in the birth, information deficits. (2) In 1995 and 2006, the birth support roles of family members (partners, sisters, mothers-in-law etc.) were documented per 1000 consecutive hospital births and analysed according to parity and ethnicity. RESULTS: (1) Structured interviews based on guidelines: In comparing 1995/96 (G/T n = 100, in each case) to 2003 (G/ n = 54, T/ n = 42) there is a significant increase among both Germans and ethnic Turks in the number of fathers participating in courses preparing for birth; ethnic Turkish men attended birthing courses noticeably less often (1995/96 and 2003: 8%; G approx. 40%, respectively). During both interview cycles the most common motivation given for providing birth support was 'to solidify the relationship with the partner'. (2) Data on birth support roles: In the comparison between 1995/96 and 2006, more German fathers, as well as more ethnic Turkish fathers, were by the side of their wives/partners in the labour and delivery room; however, in contrast to the Germans, ethnic Turkish men more commonly tended to participate in a birth support role together with female family members, either simultaneously or by turns. CONCLUSIONS: The birth support role of the father has gained significant ground in the last 10 years, and is now also accepted among ethnic Turkish couples despite cultural differences. This is possibly the result of an acculturation process, in which a phenomenon within the host culture (fathers in the labour and delivery room) mixes with ancestral tradition (the birth support role of women). PMID- 19308778 TI - Depression and anxiety among Polish infertile couples--an evaluative prevalence study. AB - AIM: The aim of this study was to evaluate the influence of infertility on the severity of anxiety and depression in infertile couples. MATERIAL AND METHODS: This was a cross-sectional study of differences between infertile couples (206 women and 188 men) and fertile couples (n = 190) with symptoms of depression and anxiety, as measured by the Beck Depression Inventory and Beck Anxiety Inventory. RESULTS: Infertile women (35.44%) scored above the cut-off for severe symptoms of depression, compared with 19.47% of fertile women. In the case of anxiety evaluation there was significant total prevalence among infertile women (15.53%). In the male groups there was a comparable frequency of negative results for depression and anxiety and their intensity. Among Female Infertile, depression occurred most frequently in combined infertility, whilst among Male Infertile in male infertility, with a time-frame of 3-6 years causing the creation and severity of depressive symptoms. CONCLUSION: The risk factors of depression and anxiety in infertility include: female sex, age over 30, lower level of education, lack of occupational activity, diagnosed male infertility and infertility duration of 3-6 years. PMID- 19308779 TI - The role of maternal anxiety in the early postpartum period: screening for anxiety and depressive symptomatology in Greece. AB - BACKGROUND: From birth to the first year postpartum, there is a critical period for the development of affective disorders. Maternal anxiety has received little attention even though it is associated with a number of adverse outcomes. Symptoms of anxiety often comorbid with depression and pertain a significant role in the maintenance of postpartum distress. The purpose of this study is to assess anxiety and depressive symptomatology in a Greek population and to examine their relationship. This study investigated the demographic and socio-psychological factors that are associated with the onset of the symptoms of postpartum distress. METHOD: The study was conducted at the perinatal hospital Elena Venizelou in Greece. Two hundred thirty-five mothers met the inclusion criteria and participated in the study. The state-trait inventory was administered to screen symptoms of anxiety. It incorporates the state subscale that measures symptoms of temporal anxiety, and trait subscale that measures personality predisposition to anxiety. The Edinburgh postpartum depression scale (EPDS) inventory was administered to screen for symptoms of depression. The first assessment was conducted in 2-3 days after labor and the follow-up assessment was conducted in 3 months postpartum by telephone. A standard survey questionnaire was used for the purposes of collecting the demographic data. RESULTS: Symptoms of postpartum depression had 14.5% of mothers on the first screening and 4.6% at the follow-up (EPDS >or= 14). State anxiety symptoms were manifested by 22.9% of the sample on the first screening and 12.6% at the follow-up. Trait anxiety symptoms exhibited 24.6% of the sample on the first screening and 14.3% at the follow-up. There was comorbidity between the symptoms of anxiety and depression. The comorbidity was evident even when the anxiety subscale of the EPDS was removed. State anxiety was correlated with primiparity, admission to the NICU and negative experience of labor. Trait anxiety was correlated with the marital status of the mother. The symptoms of depression were correlated with the young age of the mother and negative experience of labor. CONCLUSION: Symptoms of maternal anxiety are common after labor in Greece and persist in the early postpartum period. This finding suggests that the impact of maternal anxiety should be considered when studying postpartum distress. The comorbidity amongst anxiety and depressive symptomatology persisted at 3 months postpartum making women more vulnerable to postpartum distress. Given this result screening prior to hospital discharge is essential as it can provide an indication of the mothers who are susceptible to developing affective disorders. PMID- 19308780 TI - Sense of coherence predicts uncomplicated delivery: a prospective observational study. AB - The present study aimed to investigate factors predicting uncomplicated deliveries and specifically whether a sense of coherence (SOC) and perceived stress can predict such deliveries. A prospective observational study was conducted employing self-administered SOC and perceived stress scale (PSS) questionnaires with pregnant women attending the outpatient clinic for routine surveillance. Following delivery, data regarding maternal and fetal delivery complications were collected from the participants' medical records. Of one hundred forty-five women completing the study, 43.4% completed the delivery process without complications. Women experiencing delivery complications, on average, had lower SOC scores (67.7 +/- 1.19 vs. 72.2 +/- 1.32, p = 0.014). Maternal complications (as opposed to fetal complications) accounted for this divergence and were related to lower SOC scores (67.74 +/- 1.19 vs. 72.18 +/- 1.32, p = 0.01). PSS was not associated with uncomplicated delivery (18.82 +/- 0.59 vs. 17.98 +/- 0.62, p = 0.341). Nulliparity, however, was associated with higher occurrence of complicated delivery (31.9% of complicated vs. 13.2% of uncomplicated deliveries, p = 0.007). Multivariable analysis demonstrated that high SOC (OR = 1.042; 95% CI = 1.004-1.08; p = 0.03) and nulliparity (OR = 0.293; 95% CI = 0.113-0.758; p = 0.011) both were independent predictors of uncomplicated delivery, directly and inversely, respectively. In conclusion, higher SOC scores are an independent protective factor for the prediction of uncomplicated delivery. PMID- 19308781 TI - Women who are well informed about prenatal genetic screening delay emotional attachment to their fetus. AB - BACKGROUND: Prenatal maternal serum screening allows assessment of risk of chromosomal abnormalities in the fetus and is increasingly being offered to all women regardless of age or prior risk. However ensuring informed choice to participate in screening is difficult and the psychological implications of making an informed decision are uncertain. The aim of this study was to compare the growth of maternal-fetal emotional attachment in groups of women whose decisions about participation in screening were informed or not informed. METHODS: A prospective longitudinal design was used. English speaking women were recruited in antenatal clinics prior to the offer of second trimester maternal screening. Three self-report questionnaires completed over the course of pregnancy used validated measures of informed choice and maternal-fetal emotional attachment. Attachment scores throughout pregnancy in informed and not-informed groups were compared in repeated measures analysis. RESULTS: 134 completed the first assessment (recruitment 73%) and 68 (58%) provided compete data. The informed group had significantly lower attachment scores (p = 0.023) than the not informed group prior to testing, but scores were similar (p = 0.482) after test results were known. CONCLUSION: The findings raise questions about the impact of delayed maternal-fetal attachment and appropriate interventions to facilitate informed choice to participate in screening. PMID- 19308782 TI - Parent's attitudes towards full-scale prenatal testing for genetic disorders. AB - OBJECTIVES: Innovations in the field of prenatal diagnostic testing have led to the development of molecular tests that allow the rapid detection of specific genetic defects, such as Down syndrome. In addition, full-scale tests have been developed allowing the detection of many genetic disorders in a single test. Here we examined the attitudes of pregnant women in low risk pregnancies towards full scale genetic testing and explored relationships between demographic characteristics and the level of interest. METHODS: A prospective study was performed on 115 consecutive pregnant women. They completed the same structured questionnaire at two different time points, before counselling (T1) and after 4 weeks (T2), to assess a possible change of attitude. RESULTS: At T1, 33% of the respondents were in favour of full-scale testing of their unborn child, whereas at T2, this percentage had dropped to 18%. Except for educational level, no significant relationships were noted between the demographic variables and the wish to opt for full-scale testing. A low educational level was significantly related to the interest in full-scale testing. CONCLUSIONS: Low risk pregnant women expressed little interest in full-scale genetic testing. Educational level appeared to affect their views. PMID- 19308783 TI - Information for better or for worse: interviews with parents when their foetus was found to have choroid plexus cysts at a routine second trimester ultrasound. AB - The aim of the study was to gain a theoretical understanding of parents' experiences and handling of the situation, when their foetus was diagnosed as having choroid plexus cysts, at a routine second trimester ultrasound examination. Nine couples and one mother were interviewed using one open question. Analysis method was Grounded Theory. The main concern was anxiety and the core category became need for knowledge. The other categories were frightening and confusing, judging risk and making a choice and comforting. The parents felt information during the ultrasound examination was insufficient. The time delay between the diagnosis and the doctor's appointment was also often criticized. Most of the parents in this study wanted to know what can be diagnosed by ultrasound, even if there is a small risk that the child will have a malformation or chromosome abnormality. However, when the diagnosis is made, they need adequate information, otherwise unnecessary anxiety arises. By giving sufficient information without days of delay, anxiety can hopefully be minimized. Some written information was also requested. It is of utmost importance that the staff use the same terminology and the correct name of the soft marker to the parents. PMID- 19308784 TI - Cognitive function after pre-eclampsia: an explorative study. AB - BACKGROUND: Pre-eclampsia and eclampsia relate to cerebral damage. Memory and concentration problems are frequently reported after these pregnancy-related vascular complications. We tested the hypothesis that in formerly pre-eclamptic women cognitive functioning is impaired as compared with healthy parous controls. METHODS: Women with a recent history of pre-term pre-eclampsia (PPE; n = 47), term pre-eclampsia (TPE; N = 18), pre-term birth (PBI; n = 32) or an uneventful pregnancy (UPR; n = 72) completed a set of questionnaires, measuring cognitive problems, PTSD, depression, anxiety and fatigue. In addition, in PPE and UPR participants we tested neuropsychological performance. RESULTS: PBI is related to higher levels of post-traumatic distress symptoms, which in turn diminished the neuropsychological test performance of PPE women. Nonetheless, women in the PPE and TPE groups did not report more cognitive problems than women in the PBI and UPR groups, but PPE raised the need for psychosocial cared. CONCLUSIONS: Cognitive complaints are common amongst young mothers. When tested, only those with psychological co-morbidity have neuropsychological impairment. PMID- 19308785 TI - Psychological treatment of women with psychological complaints after pre eclampsia. AB - AIM: Pre-eclampsia (PE), eclampsia (E) and/or hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome cause major perinatal and maternal morbidity, mortality and have extensive psychosocial impact. The aim of this study is to describe psychological consequences, interventions and outcomes in a population of women from a tertiary centre. METHODS: From February 2004 through April 2007, 25/141 (18%) women with a history of PE, E and/or HELLP syndrome were referred to the medical psychologist. Obstetrical history, reason for referral to medical psychologist, medical psychological conclusion after intake, treatment and outcome were evaluated. RESULTS: Twenty-four women were reached for informed consent. Their referral was mainly for dysfunctional coping. A fifth (5/24) had posttraumatic stress disorder. Most women were Caucasian primiparous who delivered preterm by ceserean section of growth-restricted infants. Twenty-two women received therapy, one was referred to a medical social worker, one did not need therapy. The main interventions were psycho-education (n = 18), supportive techniques (n = 10), increasing autonomy techniques (n = 8) and eye movement desensitisation and reprocessing (n = 7). Duration between PE and consultation of medical psychologist was significantly related to the number of sessions (p < 0.01, Pearson correlation = 0.609). CONCLUSION: Psychological treatment in cases of psychological complaints increases coping possibilities in women after exposure to PE and timely recognition reduces the treatment duration. PMID- 19308786 TI - Chronic stress in women with localised provoked vulvodynia. AB - BACKGROUND: Localised provoked vulvodynia (former vulvar vestibulitis syndrome) is the most common cause of superficial dyspareunia among young women. In a clinical setting, it is obvious that many women with localised provoked vulvodynia show signs of chronic stress. The objective of this study was to compare chronic stress in a group of women with localised provoked vulvodynia, referred to the vulvar unit at the Division of Obstetrics and Gynecology, Danderyd Hospital, Stockholm, Sweden with control women of the same age and educational level, using two independent methods. METHODS: Forty-three women with localised provoked vulvodynia and 35 healthy control women underwent a gynecological examination, filled in a health questionnaire, and self-collected saliva for analysis of cortisol at awakening and after 15, 30 and 45 min (morning awakening cortisol). Thirty-three patients and 28 controls completed a stress questionnaire. RESULTS: Morning awakening cortisol was blunted in women with localised provoked vulvodynia (p < 0.05), indicating chronic stress. The stress questionnaire showed more signs of burnout (p < 0.005), and emotional (p < 0.05) and bodily symptoms of stress (p < 0.005) in women with localised provoked vulvodynia compared with control subjects (students t-test). CONCLUSION: More women with localised provoked vulvodynia showed blunted morning awakening cortisol and reported more symptoms of stress compared with healthy control women of the same age. PMID- 19308787 TI - Analytical method for simultaneously measuring ex vivo drug receptor occupancy and dissociation rate: application to (R)-dimethindene occupancy of central histamine H1 receptors. AB - We introduce a novel experimental method to determine both the extent of ex vivo receptor occupancy of administered compound and its dissociation rate constant (k4). [Here, we reference k4 as the rate of offset of unlabeled ligand in convention with Motulsky and Mahan (1)]. We derived a kinetic rate equation based on the dissociation rate constant for an unlabeled compound competing for the same site as a labeled compound and describe a model to simulate fractional occupancy. To validate our model, we performed in vitro kinetics and ex vivo occupancy experiments in rat cortex with varying concentrations of (R) dimethindene, a sedating antihistamine. Brain tissue was removed at various times post oral administration, and histamine H1 receptor ligand [3H]-doxepin binding to homogenates from drug-treated or vehicle-treated rats was measured at multiple time points at room temperature. Fractional occupancy and k4 for (R)-dimethindene binding to H1 receptors were calculated by using our proposed model. Rats dosed with 30 and 60 mg/kg (R)-dimethindene showed 42% and 67% occupancy of central H1 receptors, respectively. These results were comparable to occupancy data determined by equilibrium radioligand binding. In addition, drug k4 rate determined by using our ex vivo method was equivalent to k4 determined by in vitro competition kinetics (dissociation half-life t(1/2) approximately 30 min). The outlined method can be used to assess, by simulation and experiment, occupancy for compounds based on dissociation rate constants and contributes to current efforts in drug optimization to profile antagonist efficacy in terms of its kinetic drug-target binding parameters. Data described by the method may be analyzed with commercially available software. Suggested fitting procedures are given in the appendix. PMID- 19308788 TI - Prenatal programming of sporting success: associations of digit ratio (2D:4D), a putative marker for prenatal androgen action, with world rankings in female fencers. AB - Associations of the second-to-fourth digit ratio (2D:4D), a putative marker for prenatal androgen action, and of absolute finger length, a putative marker for pubertal-adolescent androgen action, with sport performance were examined in a multinational sample of 87 world-class women epee fencers. Lower (masculinized) digit ratios correlated, although not significantly so, with better current and highest past world rankings. These correlations were significant for right-hand 2D:4D with controls for the most salient factors for 2D:4D (ethnicity) and world rankings (years of international experience, height, and weight). Longer (masculinized) fingers correlated strongly with better current and highest past world rankings; these correlations became insignificant with the same controls. Replicating previous evidence for fencers, left-handedness was much more prevalent in this sample (21%) than in the female general population, and left handers had somewhat, but not significantly so, lower 2D:4D as well as better world rankings than right-handers. These findings extend related evidence suggestive of prenatal programming of aptitude across a variety of sports, especially running and soccer. Some known extragenital effects of prenatal testosterone that contribute to the development of efficient cardiovascular systems, good visuospatial abilities, physical endurance and speed, and to the propensity for rough-and-tumble play, apparently promote sporting success in adult life. PMID- 19308789 TI - Caffeine and performance in clay target shooting. AB - Controversy surrounds the influence that caffeine has on accuracy and cognitive performance in precision activities such as shooting and archery. The aim of this study was to assess the effects of two doses of caffeine on shooting performance, reaction time, and target tracking times in the sport of clay target shooting. A randomized, double-blind, placebo-controlled design was undertaken by seven elite male shooters from the double-trap discipline. Three intervention trials (2 mg caffeine . kg(-1) body mass (BM); 4 mg caffeine . kg(-1) BM; placebo) were undertaken, in which shooters completed four rounds per trial of 50 targets per round. Performance accuracy (score) and digital video footage (for determination of reaction time and target tracking times) were gathered during competition. Data were analysed using repeated-measures analysis of variance. No differences in shooting accuracy, reaction time or target tracking times among the three intervention trials or across the four rounds within each intervention were observed (P > 0.05). The results indicate that ingestion of < or =4 mg caffeine . kg(-1) BM does not provide performance benefits to elite performers of clay target shooting in the double-trap discipline. PMID- 19308790 TI - Effects of cold-water immersion on physical performance between successive matches in high-performance junior male soccer players. AB - In this study, we investigated the effect of water immersion on physical test performance and perception of fatigue/recovery during a 4-day simulated soccer tournament. Twenty high-performance junior male soccer players (age 15.9 +/- 0.6 years) played four matches in 4 days and undertook either cold-water immersion (10 +/- 0.5 degrees C) or thermoneutral water immersion (34 +/- 0.5 degrees C) after each match. Physical performance tests (countermovement jump height, heart rate, and rating of perceived exertion after a standard 5-min run and 12 x 20-m repeated sprint test), intracellular proteins, and inflammatory markers were recorded approximately 90 min before each match and 22 h after the final match. Perceptual measures of recovery (physical, mental, leg soreness, and general fatigue) were recorded 22 h after each match. There were non-significant reductions in countermovement jump height (1.7-7.3%, P = 0.74, eta(2) = 0.34) and repeated sprint ability (1.0-2.1%, P = 0.41, eta(2) = 0.07) over the 4-day tournament with no differences between groups. Post-shuttle run rating of perceived exertion increased over the tournament in both groups (P < 0.001, eta(2) = 0.48), whereas the perceptions of leg soreness (P = 0.004, eta(2) = 0.30) and general fatigue (P = 0.007, eta(2) = 0.12) were lower in the cold-water immersion group than the thermoneutral immersion group over the tournament. Creatine kinase (P = 0.004, eta(2) = 0.26) and lactate dehydrogenase (P < 0.001, eta(2) = 0.40) concentrations increased in both groups but there were no changes over time for any inflammatory markers. These results suggest that immediate post match cold-water immersion does not affect physical test performance or indices of muscle damage and inflammation but does reduce the perception of general fatigue and leg soreness between matches in tournaments. PMID- 19308791 TI - Voice handicap index in Swedish. AB - The objective of this study was to evaluate a Swedish version of the voice handicap index questionnaire (Sw-VHI). A total of 57 adult, dysphonic patients and 15 healthy controls completed the Sw-VHI and rated the degree of vocal fatigue and hoarseness on visual analogue scales. A perceptual voice evaluation was also performed. Test-retest reliability was analyzed in 38 subjects without voice complaints. Sw-VHI distinguished between dysphonic subjects and controls (P<0.001). The internal consistency (Cronbach's alpha > 0.84) and test-retest reliability (intraclass correlation coefficient >0.75) were good. Only moderate or weak correlations were found between Sw-VHI and the subjective and perceptual voice ratings. The data indicate that a difference above 13 points for the total Sw-VHI score and above 6 points for the Sw-VHI subscales is significant for an individual when comparing two different occasions. In conclusion, the Sw-VHI appears to be a robust instrument for assessment of the psycho-social impact of a voice disorder. However, Sw-VHI seems to, at least partly, capture different aspects of voice function to the subjective voice ratings and the perceptual voice evaluation. PMID- 19308793 TI - Protection by glycyrrhizin against warm ischemia-reperfusion-induced cellular injury and derangement of the microcirculatory blood flow in the rat liver. AB - BACKGROUND/AIM: The mechanism by which ischemia-reperfusion (I/R)-induced derangement of the hepatic microcirculation leads to tissue injury is not fully understood. We postulated that alterations to the hepatic microcirculation, including hemodynamic derangement and increased leukocyte-endothelium interaction, play a role, and that glycyrrhizin exerts its hepatoprotective effects, in part, by reducing these microcirculatory changes. MATERIALS AND METHODS: Wistar rats were subjected to 30-60 minutes segmental hepatic ischemia, followed by 120 minutes of reperfusion. Glycyrrhizin was administered prior to ischemia. Using intravital fluorescence microscopy, the administration of fluorescein isothiocyanate-conjugated erythrocytes allowed the measurement of erythrocyte-velocity (RBC(vel)), lobular, and sinusoidal perfusion. Bleb formation was observed by electron microscopy. Blood and tissue were taken for the assessment of liver injury. RESULTS: Glycyrrhizin reduced I/R-induced liver injury (histology, liver enzymes) and reduced hepatocyte apoptosis (TUNEL, caspase-3 activity). Glycyrrhizin inhibited hepatocyte bleb formation and reversed the I/R-induced reductions in lobular perfusion and RBC(vel). Leukocyte rolling and adherence in postsinusoidal venules and neutrophil infiltration were reduced by glycyrrhizin. I/R-induced elevation in HMGB1 was prevented by glycyrrhizin. CONCLUSIONS: Early bleb formation with deranged microcirculatory flow and leukocyte-endothelium interaction would appear to contribute to I/R induced hepatocellular injury. Glycyrrhizin exerts its hepatoprotective effect by preventing these changes, in addition to a direct cellular effect. PMID- 19308794 TI - Sex-related functional asymmetry of the amygdala: preliminary evidence using a case-matched lesion approach. AB - We have reported previously that there appears to be an intriguing sex-related functional asymmetry of the prefrontal cortices, especially the ventromedial sector, in regard to social conduct, emotional processing, and decision-making, whereby the right-sided sector is important in men but not women and the left sided sector is important in women but not men. The amygdala is another structure that has been widely implicated in emotion processing and social decision-making, and the question arises as to whether the amygdala, in a manner akin to what has been observed for the prefrontal cortex, might have sex-related functional asymmetry in regard to social and emotional functions. A preliminary test of this question was carried out in the current study, where we used a case-matched lesion approach and contrasted a pair of men cases and a pair of women cases, where in each pair one patient had left amygdala damage and the other had right amygdala damage. We investigated the domains of social conduct, emotional processing and personality, and decision-making. The results provide support for the notion that there is sex-related functional asymmetry of the amygdala in regard to these functions - in the male pair, the patient with right-sided amygdala damage was impaired in these functions, and the patient with left-sided amygdala damage was not, whereas in the female pair, the opposite pattern obtained, with the left-sided woman being impaired and the right-sided woman being unimpaired. These data provide preliminary support for the notion that sex related functional asymmetry of the amygdala may entail functions such as social conduct, emotional processing, and decision-making, a finding that in turn could reflect (as either a cause or effect) differences in the manner in which men and women apprehend, process, and execute emotion-related information. PMID- 19308795 TI - Old people reporting childhood AD/HD symptoms: retrospectively self-rated AD/HD symptoms in a population-based Swedish sample aged 65-80. AB - Our knowledge of attention deficit/hyperactivity disorder (AD/HD) has increased in recent years. Little is still known about the course and manifestations in later parts of life and whether elderly persons who once presented childhood AD/HD symptoms can be identified. The aim of the study was to explore the occurrence to which elderly individuals retrospectively report symptoms that may indicate childhood AD/HD. The 25-item Wender Utah Rating Scale (WURS) was administered in a population-based sample of 2500 persons aged 65-80. Demographics, self-ratings of problems in childhood, current health and memory were also investigated. A total of 1599 individuals participated, which corresponds to a response rate of 64%. The prevalence of self-rated childhood AD/HD symptoms was 3.3% using a cut-off score of 36 or more in the WURS. Men rated significantly more AD/HD symptoms. Those who reported more childhood AD/HD symptoms also claimed general problems in childhood as well as worse current health. The proportion of individuals among 65-80-year-olds, who report childhood AD/HD symptoms is slightly lower but comparable with recent prevalence rates of childhood AD/HD. The study encourages further studies of AD/HD using a lifespan perspective. PMID- 19308796 TI - The effectiveness of technology-based patient education on self-reported deprivation of liberty among people with severe mental illness: a randomized controlled trial. AB - Deprivation of liberty (DL) in psychiatric inpatient care is common worldwide. As liberty is a central element of patients' rights, there is a need to develop most effective methods supporting patients' personal liberty. The article presents initial results from a study to determine the effectiveness of an information technology (IT)-based patient education programme on patients' experiences of being deprived of their liberty during their in-hospital stay. An overall sample of 311 patients with schizophrenia spectrum psychosis was randomized into three groups: an intervention group with needs-based computerized patient education, a patient education group with conventional education and a control group with standard care. Data on the general experience of DL were collected at baseline and during the patient discharge process. In general, all patients experienced less DL at the time of their discharge. The change in patients' experiences of their DL did not differ statistically between the three groups. Male patients in the standard care group were significantly more likely to drop out of the study than female patients. Although technology-based patient education was not found to be superior to other approaches, we did not find any reason to inhibit its utilization in patient care among persons with severe mental health problems. From the healthcare organizations' perspective, a cost-effectiveness analysis is needed, as the IT education was slightly more time-consuming. PMID- 19308797 TI - Clinical features and long-term survival in chronic intestinal pseudo-obstruction and enteric dysmotility. AB - OBJECTIVE: Chronic intestinal pseudo-obstruction (CIP) is the most severe form of intestinal dysmotility. Enteric dysmotility (ED) has been proposed as a new diagnostic label for patients with disturbed intestinal motility and severe symptoms but no radiological signs of pseudo-obstruction. The purpose of this study was to compare the clinical features, small-bowel manometry findings and long-term survival in patients with CIP and ED. MATERIAL AND METHODS: Data collected during a 16-year period from 1987 to 2002 were retrospectively analysed and followed-up through 2007 in a tertiary referral centre. The study comprised 55 patients (41 F, median age 42 years, range 23-76) with CIP and 70 patients (63 F, median age 39 years, range 18-71) with ED. RESULTS: The median observation time was 9.9 years (range 5.2-20.1). Nineteen patients with CIP (35%) and 9 patients with ED (13%) died. Survival among patients with ED was significantly better (p<0.05). Patients with CIP (49%) needed parenteral nutrition more often than patients with ED (14%). Small-bowel manometry showed similar abnormalities in the two groups but absence of a fed motor response to meals was seen in 16/43 patients with CIP compared to none with ED (p<0.001), sustained periods of uncoordinated phasic activity were more common (p<0.05) in CIP patients (23/45) than in ED patients (19/70) and severe hypomotility was only seen in 7 patients with CIP. CONCLUSIONS: CIP and ED differ with respect to severity of measurable physiological derangement, nutritional needs and long-term prognosis. Our findings indicate that CIP and ED are different entities that require different approaches to management. PMID- 19308798 TI - Functional status, health-related quality of life and symptom severity in patients with chronic intestinal pseudo-obstruction and enteric dysmotility. AB - OBJECTIVE: To investigate whether patients with chronic intestinal pseudo obstruction (CIP) differ from those with enteric dysmotility (ED) regarding self reported measures of functional status, health-related quality of life (HRQoL) and gastrointestinal symptoms. MATERIAL AND METHODS: The study comprised 28 patients with CIP (median age 48, range 28-80 years) and 26 with ED (median age 50, range 20-75 years). Three self-administered questionnaires were used: the Sickness Impact Profile (SIP), the Swedish HRQoL Questionnaire (SWED-QUAL) and the Gastrointestinal Symptom Rating Scale (GSRS). RESULTS: Patients with CIP reported significantly greater functional impairment in the aggregated physical dimension of SIP (15.4 SD 19.7) than did patients with ED (5.0 SD 7.2, p<0.01). They also reported significantly poorer general health (SWED-QUAL) (28.0 SD 20.8) compared to those with ED (44.8 SD 25.2, p <0.01). Symptom severity correlated with several measures of HRQoL and functional status in patients with CIP but abdominal pain severity was the only independent predictor of HRQoL. Abdominal pain alone explained between 21% and 67% of the variance in SWED-QUAL subscales. Likewise, the severity of indigestion symptoms among patients with CIP explained 20-24% of the variance in the SIP subscales, emotional behaviour and work. Much less correlation between symptom severity and functional impairment or HRQoL was found in patients with ED. We found no difference in symptom severity (GSRS) between the two groups. CONCLUSIONS: Patients with CIP reported a greater impairment of functional status and HRQoL than did patients with ED. Symptom severity had a stronger influence on functional status and HRQoL in patients with CIP. PMID- 19308799 TI - Thylakoids promote release of the satiety hormone cholecystokinin while reducing insulin in healthy humans. AB - OBJECTIVE: The effects of a promising new appetite suppressor named "thylakoids" (membrane proteins derived from spinach leaves) were examined in a single meal in man. Thylakoids inhibit the lipase/colipase hydrolysis of triacylglycerols in vitro and suppress food intake, decrease body-weight gain and raise the satiety hormone cholecystokinin (CCK) in rats, but their effects in man remain unclear. The aim of this study was to investigate whether thylakoids, when added to a test meal, affect appetite regulation and blood parameters in healthy individuals. MATERIAL AND METHODS: In an intervention crossover study, healthy individuals of normal weight (n=11) were offered a high-fat meal with and without the addition of thylakoids. Blood samples were taken 0 (prior to meal), 30, 60, 120, 180, 240, 300 and 360 min after the start of the meal. Blood samples were analysed for satiety and hunger hormones (CCK, leptin and ghrelin), insulin and blood metabolites (glucose and free fatty acids). RESULTS: The CCK level increased, in particular between the 120 min time-point and onwards, the ghrelin level was reduced at 120 min and leptin level increased at 360 min after intake of the thylakoid-enriched meal. The insulin level was reduced, whereas glucose concentrations were unchanged. Free fatty acids were reduced between time-point 120 min and onwards after the thylakoid meal. CONCLUSIONS: The addition of thylakoids to energy-dense food promotes satiety signals and reduces insulin response during a single meal in man. PMID- 19308800 TI - Bacteria, biofilm and honey: a study of the effects of honey on 'planktonic' and biofilm-embedded chronic wound bacteria. AB - Chronically infected wounds are a costly source of suffering. An important factor in the failure of a sore to heal is the presence of multiple species of bacteria, living cooperatively in highly organized biofilms. The biofilm protects the bacteria from antibiotic therapy and the patient's immune response. Honey has been used as a wound treatment for millennia. The components responsible for its antibacterial properties are now being elucidated. The study aimed to determine the effects of different concentrations of 'Medihoney' therapeutic honey and Norwegian Forest Honey 1) on the real-time growth of typical chronic wound bacteria; 2) on biofilm formation; and 3) on the same bacteria already embedded in biofilm. Reference strains of MRSE, MRSA, ESBL Klebsiella pneumoniae and Pseudomonas aeruginosa were incubated with dilution series of the honeys in microtitre plates for 20 h. Growth of the bacteria was assessed by measuring optical density every 10 min. Growth curves, biofilm formation and minimum bactericidal concentrations are presented. Both honeys were bactericidal against all the strains of bacteria. Biofilm was penetrated by biocidal substances in honey. Reintroduction of honey as a conventional wound treatment may help improve individual wound care, prevent invasive infections, eliminate colonization, interrupt outbreaks and thereby preserve current antibiotic stocks. PMID- 19308801 TI - An outbreak of human metapneumovirus infection in hospitalized psychiatric adult patients in Taiwan. AB - Human metapneumovirus (hMPV) is a paramyxovirus that is associated with respiratory tract infection (RTI) mostly in children, but these outbreaks have rarely been reported in adults. We encountered an outbreak of this disease involving 10 adults in a psychiatric ward in eastern Taiwan. The nasopharyngeal swab specimens from 13 patients with symptoms of RTI were obtained and analyzed. The RT-PCR tests were negative to influenza virus A/B, adenovirus, RSV, parainfluenza virus, coronavirus, Nipah virus and Legionella. The antigen tests were negative to Legionella, Chlamydia, and Mycoplasma. Blood culture was negative in all except patient no. 1, who was found positive for coagulase negative staphylococci. The hMPV was identified in 10 of 13 adults (77%), but negative for the other virus. Cough was present in all (100%), fever in 90%, and X-ray evidence of pneumonia in 7 patients. One patient died of respiratory failure. We report this outbreak in a mental hospital to alert the medical profession that this unusual infection of hMPV can occur as an outbreak in an adult setting and is an occupational hazard for healthcare personnel. PMID- 19308802 TI - Lipoprotein(a) in late onset neonatal sepsis. PMID- 19308803 TI - Reproductive health in women with systemic lupus erythematosus compared to population controls. AB - OBJECTIVE: To study the reproductive health history in women with systemic lupus erythematosus (SLE) compared to population controls. METHODS: A total of 206 female SLE patients were interviewed regarding demographic and disease data, menstruation, use of contraception and hormone replacement therapy (HRT), infertility, and pregnancies. The control group consisted of 1037 women from the general population of similar age and socioeconomic status living in the same region. RESULTS: In SLE women compared to population controls, mean age at menarche (13.3 vs. 13.2 years) and frequency of infertility (16% vs. 16%) were similar but menopause occurred earlier (44.9 vs. 46.8 years, p = 0.01). Current use of oral contraceptives (OCs) was less common than in controls [18% vs. 28%, odds ratio (OR) 0.55, 95% CI 0.3-1.0] while previous use of progesterone containing intrauterine devices (IUDs) was more common (13% vs. 5%, OR 3.2, 95% CI 1.9-5.4). Current use of HRT was similar (22% vs. 21%) but SLE patients had started the use earlier (43.2 vs. 47.1 years, p = 0.003). Mean number of pregnancies was lower in SLE patients compared to controls (2.3 vs. 2.5, p = 0.046) and in lupus nephritis patients compared to SLE patients without nephritis (1.9 vs. 2.5, p = 0.01). No difference was found in the occurrence of spontaneous and induced abortions compared to controls, but pregnancy-associated complications were more common in SLE women. CONCLUSION: When compared to population controls women with SLE are normally fertile, use less OCs and more IUDs, have earlier menopause and use HRT as frequently. Family size is reduced, especially in lupus nephritis patients, and pregnancy-associated complications are more common. PMID- 19308804 TI - On the recurrence risk of arthritis among psoriatic patients. PMID- 19308806 TI - Bullous pemphigoid is a rare paraneoplastic syndrome in patients with renal cell carcinoma. AB - Renal cell carcinoma is associated with paraneoplastic syndromes in up to 40% of cases. Dermatological manifestations are rare. A case of a bullous pemphigoid as a paraneoplastic symptom was diagnosed in a 52-year-old patient with a partially sarcomatoid papillary renal cell carcinoma. PMID- 19308805 TI - Incidence of cancer among patients with systemic sclerosis in Korea: results from a single centre. AB - OBJECTIVE: The aim of this study was to determine whether the incidence of cancer has increased among patients with systemic sclerosis (SSc) in Korea. METHODS: The study subjects consisted of 112 patients who had been consecutively evaluated for at least 6 months between 1990 and 2007. We retrospectively reviewed their medical records, investigated the incidence rate of cancer and compared it with that of the Korea National Cancer Centre database. RESULTS: Nine out of 112 patients developed cancer (four males and five females). The average age at diagnosis of cancer was 56.4 years and the mean disease duration was 8.9 years. The standardized incidence ratio (SIR) for SSc patients was 4.2 [95% confidence interval (CI) 2.3-6.1], 3.7 for women (95% CI 1.2-6.2) and 6.4 for men (95% CI 1.6-11.2). Lung cancer was the most common cancer (n = 4), followed by oesophagus (n = 1), stomach (n = 1), liver (n = 1), pancreas (n = 1), and squamous cell carcinoma of unknown origin (n = 1). All patients who developed lung cancer were female and non-small cell carcinoma in origin. The SIR of lung cancer in female patients was 23.0 (95% CI 6.0-40.0). Two out of four lung cancer patients had concomitant interstitial lung disease (ILD); all were non-smokers. Treatment agents, autoantibodies, smoking, and lung involvement were not significantly different between SSc patients with or without cancer. CONCLUSION: The SIR of cancer was significantly higher in SSc patients, and especially in those who were male, than in the general population. Lung cancer was the most common cancer. Active surveillance for the detection of cancer should be performed in all SSc patients. PMID- 19308807 TI - Therapeutic effects of add-on botulinum toxin A on patients with large benign prostatic hyperplasia and unsatisfactory response to combined medical therapy. AB - OBJECTIVE: Botulinum toxin A (BoNT-A) injections into the prostate have been reported to be effective and durable in the treatment of small benign prostatic hyperplasia (BPH). This study evaluated the effectiveness of BoNT-A in patients with large BPH with an unsatisfactory response to combined alpha-blocker and 5 alpha-reductase inhibitor therapy. MATERIAL AND METHODS: Sixty patients with BPH and total prostate volume (TPV) of > 60 ml with unsatisfactory response to combination medical therapy were randomly assigned to receive add-on intraprostatic BoNT-A injection (n=30) or continued medical therapy (control group). Patients in the treatment group received 200-600 U of Botox injected into the prostate. Outcome parameters including International Prostate Symptom Score (IPSS), quality of life index (QoL-I), TPV, maximum flow rate (Q(max)) and postvoid residual (PVR) were compared between treatment and control groups at baseline, 6 months and 12 months. RESULTS: Significant decreases in IPSS, QoL-I and TPV, and increase in Q(max) were observed at 6 months and remained stable at 12 months in the treatment group. Improvements in IPSS and QoL-I were also observed at 6 months and a decrease in TPV at 12 months was noted in the control group. However, no significant changes in any parameters except for QoL-I at 6 and 12 months were noted between the treatment and control groups. Acute urinary retention developed in three patients receiving BoNT-A treatment. Three BoNT-A and two medical treatment patients converted to transurethral surgery at the end of study. CONCLUSIONS: This study shows that add-on prostatic BoNT-A medical treatment can reduce prostate volume and improve lower urinary tract symptom score and QoL-I within 6 months in the treatment of large BPH. However, the therapeutic effect at 12 months was similar to combination medical treatment. PMID- 19308808 TI - Urinary incontinence in women is not exclusively a medical problem: a population based study on urinary incontinence and general living conditions. AB - OBJECTIVE: The aim of the study was to analyse differences in general health and general living conditions between women with and without urinary incontinence (UI). MATERIAL AND METHODS: This cross-sectional population-based study was conducted in Orebro County, Sweden. A public health questionnaire, "Life and Health", was sent to a randomly selected sample of the population. The questionnaire consisted of 87 questions on broad aspects of general and psychiatric health. An additional questionnaire was enclosed for those respondents who reported experiencing UI. The data were analysed using binary logistic regression. The final study population constituted 4609 women, 1332 of whom had completed both questionnaires. The remaining 3277 had completed only the Life and Health questionnaire. Effect measures were odds ratios (ORs) with corresponding 95% confidence intervals (CIs). RESULTS: Statistically significant associations were found between UI and the occurrence of musculoskeletal pain (OR 1.45, 95% CI 1.20-1.76), fatigue and sleeping disorders (OR 1.59, 95% CI 1.30 1.95), feelings of humiliation (OR 1.29, 95% CI 1.12-1.50), financial problems (OR 1.36, 95% CI 1.11-1.66), and reluctance to seek medical care (OR 1.43, 95% CI 1.21-1.68). CONCLUSION: UI among women is commonly associated with a number of different psychosocial problems as well as an expressed feeling of vulnerability. PMID- 19308810 TI - Psychosocial risk factors for depression during pregnancy. AB - OBJECTIVE: To assess the prevalence of antenatal depressive disorder in different trimesters and to evaluate the relation of psychosocial risk factors to antenatal depressive disorder. DESIGN: Cohort follow-up. SETTING: University Hospital, Kaunas, Lithuania. SAMPLE: Two hundred and thirty pregnant women consecutively admitted. METHODS: At 12-16 weeks, 22-26 weeks, and 32-36 weeks of pregnancy, participants were screened for depression using the World Health Organization's Composite International Diagnostic Interview Short Form (CIDI-SF). Women who gave at least one positive answer to the CIDI-SF depression-screening question were evaluated for depressive disorder using the non-patient version of the Structured Clinical Interview for DSM-III-R (SCID-NP). Psychosocial stressors and two Big Five Personality dimensions, neuroticism and extraversion, were also evaluated. MAIN OUTCOME MEASURES: Prevalence of depressive disorder. RESULTS: The prevalence of the antenatal depressive disorder at 12-16 weeks' gestation was 6.1%, at 22-26 weeks 3.5%, and at 32-36 weeks 4.4%. In the first trimester, a greater prevalence of current depressive disorder was independently associated with unplanned and unwanted pregnancy, high neuroticism, low education, and a previous history of depression; in the second trimester with unplanned and unwanted pregnancy and high neuroticism; in the third trimester with unplanned and unwanted pregnancy, high neuroticism, and the occurrence of psychosocial stressors during the last year. CONCLUSIONS: The highest prevalence of depressive disorders was found in the first trimester, the lowest in mid-pregnancy. Several determinants (unwanted and unplanned pregnancy, high neuroticism) were independent predictors of antenatal depressive disorders throughout whole pregnancy, while other determinants (low education, previous history of depression, the occurrence of psychosocial stressors at the end of pregnancy) were trimester specific. PMID- 19308811 TI - Skin-limited Langerhans' cell histiocytosis in children. AB - Langerhans' cells are dendritic cells derived from precursors in the bone marrow. They constitute 2-4% of the resident epidermal cells and are found within the epidermis above the basal layer. They function as immunologic cells by recognizing antigens and presenting them to T cell lymphocytes. Langerhans' cell histiocytosis is a rare pathology characterized by an abnormal clonal proliferation of Langerhans' cells that infiltrates different organs of the human body. The proliferating Langerhans' cells appears to be primarily responsible for the clinical manifestations. The stimulus for their proliferation is unknown. Among different organs, cutaneous involvement is encountered in 40% of cases. The aim of this investigation is to review the clinicopathologic, immunologic and ultrastructural features of skin-confined Langerhans' cell histiocytosis in children through seven case series. Four boys and two girls with age range of 1 year to 8 years presented with scaling, crusted papules, nodules and papulonodular lesions (two cases each). The locations included the face (three cases), scalp, trunk and vulva (one case each). The histological features included histiocytic reaction (one case), granulomatous reactions (three cases) and both granulomatous and histiocytic reactions (two cases). The diagnosis was confirmed by histochemical (S-100 + CD1a +) and ultrastructural studies (Birbeck granules). Langerhans' cell histiocytosis is a rare disease with pleomorphic cutaneous clinical expressions. Three types of skin lesions usually occur: nodules (common), scaling, or crusted papules (next in frequency) and finally soft, yellow papular xanthomas (rare). Three types of histological pictures are seen: histiocytic, granulomatous (common) and xanthomatous (rare). PMID- 19308812 TI - Single nucleotide polymorphisms in human gene icb-1 and breast cancer susceptibility. AB - In this study, we tested the hypothesis that single nucleotide polymorphisms (SNPs) of differentiation-associated human gene icb-1 (C1orf38) may be associated with breast cancer susceptibility. A total of 646 women--323 breast cancer cases and just as many controls--were included. Breast cancer patients more frequently carried the homozygous genotype AA of SNP rs1467465 than did healthy women. Analysis of allele positivity revealed that AG or GG genotypes were significantly less frequent in breast cancer patients, suggesting that presence of G allele might have protective effects. Our data suggest that SNP rs1467465 of human gene icb-1 might affect breast cancer susceptibility. PMID- 19308813 TI - t(3;9;22) 3-way chromosome translocation in chronic myeloid leukemia is associated with poor prognosis. AB - Chronic myelogenous leukemia (CML) is genetically characterized by the reciprocal translocation of chromosome 9 and 22. Around 5-8% of CML develop complex variant Ph translocations involving one or more chromosomal regions besides 9 and 22. Chromosome 3 is not frequently involved in complex translocations in CML. We report in this study a case of CML displaying a t(3;9;22) 3-way translocation. A review of the literature appears to indicate that CML patients with this translocation tend to have an aggressive course and poor outcome. Additional 3 way chromosome translocations associated with CML are also reviewed. PMID- 19308814 TI - Festschrift for Dr Don B. Chaffin. PMID- 19308815 TI - The evolving role of biomechanics in prevention of overexertion injuries. AB - This paper describes occupational biomechanics as an evolving body of knowledge that has required not only a sophisticated development of fundamental biomechanical principles and human failure data, but also has required epidemiological information to enable a more complete understanding of how certain types of musculoskeletal injuries can be caused by specific physical work requirements. It also is argued that even with adequate biomechanical and epidemiological information, the ability to change working conditions and manual task requirements in companies required management and workers to become organised into formal ergonomics teams that could be trained and empowered to reduce the known biomechanical risk factors present in various jobs. It is demonstrated that in the last 35 years occupational biomechanics research continues to provide the intellectual machine that is driving the development of important ergonomics guidelines. Despite these successes, however, some major limitations in contemporary biomechanics knowledge are discussed, particularly related to situations where high-speed motions and repetitions are involved. Finally, the evolving importance and limitations in occupational biomechanical simulation models for proactive ergonomics are presented. PMID- 19308816 TI - Prolonged mechanical and physiological changes in the upper extremity following short-term simulated power hand tool use. AB - This study investigated in-vivo changes in upper limb dynamic mechanical properties and magnetic resonance imaging (MRI) parameters following short-term power hand tool operation. Previous studies have found reduction in mechanical properties following short-term power tool usage at long build-up times. This study advances that work by having participants operate a simulated pistol grip power hand tool and evaluating changes in mechanical properties, strength, discomfort level and MRI prior to tool operation and daily for 3 d after tool operation. Twenty-four participants were randomly assigned to operate a simulated power hand tool for either a high peak reaction force of 123 N (peak torque=8 Nm, build-up time=250 ms) or at a low peak reaction force of 5 N (peak torque=2 Nm, build-up time=50 ms). Subjects operated the tool for 60 min at the rate of six times per min. A reduction in stiffness (27%, p<0.05) was observed 24 h after tool operation for the high force group and this change persisted (26%, p<0.05) up to 72 h after tool operation. Similar changes were not observed for the low force group. No changes were observed in mass moment of inertia, damping, isometric strength and damping for either group (p>0.05). There was a signal intensity increase (12%, CI 19%, 5.06%) in the supinator muscle MRI for both groups 24 h after tool operation but only the high force group remained elevated (10%, CI 13.7%, 0.06%) 72 h after tool operation. Persistent short-term changes in mechanical and MRI parameters at high force levels could indicate increased strain on the upper limb and may negatively affect ability to react during rapid forceful loading of the upper limb. This research can ultimately lead to better ergonomic interventions through quantitative power hand tool design guidelines and work practices based on understanding the damaging effects of exposure to specific levels of reaction force, build-up time and repetition, as well as providing new outcome measures for epidemiological studies. PMID- 19308817 TI - The effect of added degrees of freedom and handle type on upper limb muscle activity during simulated hand tool use. AB - The human upper limb serves a number of functions ranging from coarse movements such as supporting a load when lifting overhead to the fine motor control required when painting a portrait. However, there are limited data available that address upper extremity function and performance when using hand tools in situations where the tool endpoint is not fixed but can move translationally or rotationally. The goal of this study was to examine variation in arm muscle activity when added degrees of freedom (DOF) were introduced through the use of a force application apparatus with two different handle designs (D-handle or screwdriver). Electromyography of seven forearm muscles and five muscles crossing the shoulder joint were measured to determine relative activity from a reference (0 DOF), most stable condition, to combinations of DOF ranging from 1 to 4. Substantial and statistically significant increases in muscle activity resulted from adding DOF. The screwdriver handle increased forearm muscle activity compared to the D-handle, except in the highest DOF condition. These findings have significance in the planning of work and design of tools because of the potential for increased fatigue that accompanies increased DOF at the tool endpoint. Handle type also influenced the magnitude of the muscular activity. PMID- 19308818 TI - Applications of biomechanics for prevention of work-related musculoskeletal disorders. AB - This paper summarises applications of biomechanical principles and models in industry to control musculoskeletal disorders of the low back and upper extremity. Applications of 2-D and 3-D biomechanical models to estimate compressive force on the low back, the strength requirements of jobs, application of guidelines for overhead work and application of strain index and threshold limit value to address distal upper extremity musculoskeletal disorders are presented. Several case studies applied in the railroad industry, manufacturing, healthcare and warehousing are presented. Finally, future developments needed for improved biomechanical applications in industry are discussed. The information presented will be of value to practising ergonomists to recognise how biomechanics has played a significant role in identifying causes of musculoskeletal disorders and controlling them in the workplace. In particular, the information presented will help practising ergonomists with how physical stresses can be objectively quantified. PMID- 19308819 TI - Spine loading at different lumbar levels during pushing and pulling. AB - As the nature of many materials handling tasks have begun to change from lifting to pushing and pulling, it is important that one understands the biomechanical nature of the risk to which the lumbar spine is exposed. Most previous assessments of push-pull tasks have employed models that may not be sensitive enough to consider the effects of the antagonistic cocontraction occurring during complex pushing and pulling motions in understanding the risk to the spine and the few that have considered the impact of cocontraction only consider spine load at one lumbar level. This study used an electromyography-assisted biomechanical model sensitive to complex motions to assess spine loadings throughout the lumbar spine as 10 males and 10 females pushed and pulled loads at three different handle heights and of three different load magnitudes. Pulling induced greater spine compressive loads than pushing, whereas the reverse was true for shear loads at the different lumbar levels. The results indicate that, under these conditions, anterior-posterior (A/P) shear loads were of sufficient magnitude to be of concern especially at the upper lumbar levels. Pushing and pulling loads equivalent to 20% of body weight appeared to be the limit of acceptable exertions, while pulling at low and medium handle heights (50% and 65% of stature) minimised A/P shear. These findings provide insight to the nature of spine loads and their potential risk to the low back during modern exertions. PMID- 19308820 TI - Does the asymmetry multiplier in the 1991 NIOSH lifting equation adequately control the biomechanical loading of the spine? AB - The aim of this research was to evaluate whether the asymmetry multiplier incorporated in the 1991 National Institute for Occupational Safety and Health lifting equation adequately controls the biomechanical spine loads during asymmetric lifting. Sixteen male subjects lifted a box from four initial locations varying in terms of the angular deviation from the mid-sagittal plane (0, 30, 60 and 90 degrees). From each location, boxes that weighed the recommended weight limit (RWL) and three times the RWL were lifted at two qualitatively defined lifting speeds. Ground reaction forces were combined with kinematic data in a linked-segment model to quantify the 3-D moments at the base of the spine (L5/S1) and the spine compression forces. The results show that the twisting and lateral bending moments increased with task asymmetry despite the lessening of the RWL (p<0.01). The flexion moment and the spine compression decreased with asymmetry, although at a slower rate than the RWL. When the dynamics were removed from the linked segment spine model to approximate the assumption of slow and smooth lifting, the estimated compression remained approximately 3400 N across all asymmetry conditions. Thus, the reduction in the RWL due to asymmetry multiplier appears appropriate and should not be changed, as been suggested by recent psychophysical studies. PMID- 19308821 TI - Evolving ergonomics? AB - The theme developed in this position paper follows the current evolution of injury prevention in the backs of workers. Job change or 'fitting the task to the person' has come far, but will probably not result in zero injury rates. This is because the cause of injury is heavily influenced by the way that a worker moves. A review of injury mechanisms reveals the need for the biomechanist/ergonomist to incorporate features in biomechanical models that recognise these injury mechanisms. The implication of one such model is that the next leap toward a zero injury rate may be approached with 'fitting the person to the task' or at least retraining the way that workers move. A few examples of movement-based back injury prevention strategies are provided. Finally, some thoughts on implementing such an approach are expressed. This is a review and position paper written in honour of Professor Don Chaffin's career. PMID- 19308822 TI - Development of a decision support system for residential construction using panellised walls: approach and preliminary results. AB - There is a high prevalence of work-related musculoskeletal disorders (WMSDs) among residential construction workers, yet control in this industry can be difficult for a number of reasons. A decision support system (DSS) is described here to allow early assessment of both ergonomic and productivity concerns, specifically by designers. Construction using prefabricated walls (panels) is the focus of current DSS development and is based conceptually on an existing 'Safety in Construction Design' model. A stepwise description of the development process is provided, including input from end users, taxonomy development and task analysis, construction worker input, detailed laboratory-based simulations and modelling/solution approaches and implementation. Preliminary results are presented for several steps. These results suggest that construction activities using panels can be efficiently represented, that some of these activities involve exposure to high levels of WMSD risk and that several assumptions are required to allow for ease of mathematical and computational implementation of the DSS. Successful development of such tools, which allow for proactive control of exposures, is argued as having substantial potential benefit. PMID- 19308823 TI - The effects of split keyboard geometry on upper body postures. AB - Split, gabled keyboard designs can prevent or improve upper extremity pain among computer users; the mechanism appears to involve the reduction of awkward wrist and forearm postures. This study evaluated the effects of changes in opening angle, slope and height (independent variables) of a gabled (14 degrees) keyboard on typing performance and upper extremity postures. Twenty-four experienced touch typists typed on seven keyboard conditions while typing speed and right and left wrist extension, ulnar deviation, forearm pronation and elbow position were measured using a motion tracking system. The lower keyboard height led to a lower elbow height (i.e. less shoulder elevation) and less wrist ulnar deviation and forearm pronation. Keyboard slope and opening angle had mixed effects on wrist extension and ulnar deviation, forearm pronation and elbow height and separation. The findings suggest that in order to optimise wrist, forearm and upper arm postures on a split, gabled keyboard, the keyboard should be set to the lowest height of the two heights tested. Keyboard slopes in the mid-range of those tested, 0 degrees to -4 degrees, provided the least wrist extension, forearm pronation and the lowest elbow height. A keyboard opening angle in the mid-range of those tested, 15 degrees, may provide the best balance between reducing ulnar deviation while not increasing forearm pronation or elbow separation. These findings may be useful in the design of computer workstations and split keyboards. The geometry of a split keyboard can influence wrist and forearm postures. The findings of this study are relevant to the positioning and adjustment of split keyboards. The findings will also be useful for engineers who design split keyboards. PMID- 19308824 TI - Comparison of required operating forces between floor-based and overhead-mounted patient lifting devices. AB - This study investigated the differences in required push, pull and rotating forces for moving fully loaded, floor-based and overhead-mounted full body patient lifting devices with simulated patients of varying weight on a floor of optimal design (i.e. level vinyl tile over concrete). A single person operated the lifting devices for all of the tests. Eighteen male and female volunteer participants, ranging in weight from 51 to 146 kg, acted as patients during the lifting tests. For each test, the simulated patients were pushed and pulled for 3.7 linear metres and were rotated while sitting in the lift slings. Force measurements were acquired using two single axis dynamometers affixed to the lifting devices. Results revealed that, in general, operator input force and torque increased with patient weight category and floor-based lifts required greater force and torque compared to the overhead-mounted lift. Comparison of the required forces with published force limits reveals that the required push and pull force from the various patient lift systems, across all weight categories, were generally acceptable to 90% of the female population. The required forces for these patient transfer tasks, however, could exceed maximum acceptable force limits if the floor surfaces were less than ideal, such as floors composed of carpet, wood, or inclined surfaces. Additional research is needed to assess these conditions. PMID- 19308825 TI - A comparison of two methods of measuring static coefficient of friction at low normal forces: a pilot study. AB - This study compares two methods for estimating static friction coefficients for skin. In the first method, referred to as the 'tilt method', a hand supporting a flat object is tilted until the object slides. The friction coefficient is estimated as the tangent of the angle of the object at the slip. The second method estimates the friction coefficient as the pull force required to begin moving a flat object over the surface of the hand, divided by object weight. Both methods were used to estimate friction coefficients for 12 subjects and three materials (cardboard, aluminium, rubber) against a flat hand and against fingertips. No differences in static friction coefficients were found between the two methods, except for that of rubber, where friction coefficient was 11% greater for the tilt method. As with previous studies, the friction coefficients varied with contact force and contact area. Static friction coefficient data are needed for analysis and design of objects that are grasped or manipulated with the hand. The tilt method described in this study can easily be used by ergonomic practitioners to estimate static friction coefficients in the field in a timely manner. PMID- 19308827 TI - The effects of nursing home placement on the perceived levels of caregiver burden. AB - Providing care for an aging parent can be one of the most fulfilling life experiences for an adult child. It can also be one of the most exhausting physically, emotionally, and financially. A caregiver experiences psychological and emotional changes when their dependent parent or spouse is placed into formal care. This research project uses the Montgomery Borgatta Caregiver Burden Scale, amended with a questionnaire, in a self-administered, anonymous survey to explore perceptions of caregiving burden before and after the nursing home placement periods. This research showed that numerous factors influence caregivers' perceptions of burden and the quality of relationships among family members. PMID- 19308828 TI - Compassion fatigue and the adult protective services social worker. AB - Compassion fatigue is a relatively new term that describes the symptoms that are experienced by social workers and other helping professionals who work with clients experiencing trauma. This article defines the concept of compassion fatigue and relates compassion fatigue to Adult Protective Services (APS) social workers. It is proposed that APS social workers may be susceptible to the deleterious effects of compassion fatigue due to the nature of their work and environment. Suggestions for avoidance of compassion fatigue are also discussed, including self-care strategies and the need for continuing education regarding this phenomenon. PMID- 19308829 TI - Impact of geriatric assessment and self-management support on community-dwelling older adults with chronic illnesses. AB - PURPOSE: The purpose of this study was to assess the effectiveness of interdisciplinary geriatric home-based assessment and self-management support services to community-dwelling older adults. DESIGN: A quasi-experimental, pre post test design tested two types of service delivery models. The first protocol included geriatric assessment services, with a brief self-management care plan intervention. The second protocol added a telephone support intervention. RESULTS: All participants showed significant progress in improved self-efficacy, self-rated health, functional status and physical mobility (specifically lower extremity muscle strength), mental health (specifically reduced depression) and in reduced fall hazards in their physical home environment. The telephone support intervention protocol did not provide additional value to the first protocol. IMPLICATIONS: This study demonstrates the value of non-medical intervention strategies for community dwelling older adults with chronic illnesses. PMID- 19308830 TI - How social workers demonstrate respect for elderly clients. AB - Although respect is a crucial aspect of social work practice, few studies have examined how social workers convey their respect for elderly clients. This study explored the various forms of respect demonstrated by social workers when they were with older clients. Fifty social workers serving elderly clients were surveyed by a questionnaire with closed- and open-ended questions. Based on data on ways the social workers respected their elderly clients, the study identified 7 forms most frequently practiced and considered most important. They are linguistic respect, care respect, acquiescent respect, salutatory respect, presentational respect, spatial respect, and consultative respect. The key expressions of these forms are introduced in quantitative data and a narrative form. These findings provide insight as to how social workers exhibit respect for elderly clients in their practice. The results of this exploration may be useful in developing a more comprehensive typology of the forms signifying respect for elderly clients. PMID- 19308831 TI - Evaluating the roles of professional geriatric care managers in maintaining the quality of life for older Americans. AB - This study examined the roles of professional geriatric care managers (PGCMs). In depth telephone surveys were completed with 19 PGCMs. Qualitative data were coded independently by two researchers. PGCMs reported that most clients and/or families seek services in response to a health crisis or because a family member was providing care at a distance. Most emphasized the importance of treating clients as autonomous decision-makers. Services described as most useful were supporting families and caregivers, being an advocate for clients, conducting initial assessments and ongoing follow up, and educating families about community resources and the financing of long term care. Regarding maintaining the quality of life for their older clients, a number of care managers described assisting clients to live independently at home, arranging for transfer to an appropriate level of care, and helping clients die peacefully with dignity. Major challenges PGCMs encountered were family conflicts and obtaining needed services. Recommendations to improve PGCM practice include increasing awareness of services, and establishing minimum professional standards through certification. PMID- 19308832 TI - Women still taking care: the experiences of older home care workers. PMID- 19308833 TI - Social work student interest in nursing home employment: a North American study. AB - Nursing homes (NHs) are a primary setting for gerontological social work; yet, little is known about social workers' career interest relative to this setting. This article reports the findings of an online survey of social work students (177 U.S. and 91 Canadian) that sought to identify personal characteristics and beliefs that influence NH employment intentions. Students who were enrolled in bachelors programs, lacked previous practice experience, and had already had a NH field placement or wanted this experience were more likely to express employment intent. The implications of these findings for social work education and research are discussed. PMID- 19308834 TI - Examining the curricula for entry-level social workers in aging-related fields. AB - The rapidly growing older population necessitates the provision of more gerontological social workers. This study evaluates the social work programs that are offered by a university in Hong Kong, based on separate surveys of alumni and employers. It also examines the effects of alumni background characteristics on perceived training adequacy. The training is found to be moderately adequate in preparing the alumni for practice in aging-related jobs, but there is a need to improve the existing curricula. Implications for the improvement of the curricula and suggestions for further studies are also discussed. PMID- 19308837 TI - Editor's introduction. Annual Review of Sex Research. PMID- 19308838 TI - A history of erotic philosophy. AB - This essay historically explores philosophical views about the nature and significance of human sexuality, starting with the Ancient Greeks and ending with late 20th-century Western philosophy. Important figures from the history of philosophy (and theology) discussed include Sappho, Plato, Aristotle, St. Augustine, St. Jerome, the Pelagians, St. Thomas Aquinas, Michel de Montaigne, Rene Descartes, Thomas Hobbes, David Hume, Immanuel Kant, Soren Kierkegaard, Arthur Schopenhauer, Jeremy Bentham, John Stuart Mill, Karl Marx, Friedrich Engels, Sigmund Freud, Jean-Paul Sartre, Simone de Beauvoir, Wilhelm Reich, and Herbert Marcuse. Contemporary philosophers whose recent work is discussed include Michel Foucault, Thomas Nagel, Roger Scruton, Karol Wojtyla (Pope John Paul II), Catharine MacKinnon, Richard Posner, and John Finnis. To show the unity of the humanities, the writings of various literary figures are incorporated into this history, including Mark Twain, Arthur Miller, James Thurber, E. B. White, Iris Murdoch, and Philip Roth. PMID- 19308839 TI - The dual control model: current status and future directions. AB - The Dual Control Model proposes that sexual responses involve an interaction between sexual excitatory and sexual inhibitory processes. The model further postulates that individuals vary in their propensity for both sexual excitation and sexual inhibition, and that such variations help us to understand much of the variability in human sexuality. The development of psychometrically validated instruments for measuring such propensities for men (Sexual Inhibition/Sexual Excitation Scales) and for women (Sexual Excitation/Sexual Inhibition Inventory for Women) is described. These measures show close to normal variability in both men and women, supporting the concept that "normal" levels of inhibition proneness are adaptive. The relevance of the model to sexual development, sexual desire, the effects of aging, sexual identity, and the relation between mood and sexuality are discussed, and the available evidence is reviewed. Particular attention is paid to gender differences and similarities in propensities for sexual excitation and inhibition. Research findings related to sexual problems, high-risk sexual behavior, and the relevance of this model to clinical management of such problems are also summarized. Last, ideas for future use and further development of the Dual Control Model are considered. PMID- 19308840 TI - Self-help therapies for sexual dysfunction. AB - Self-help approaches can be relevant for individuals with sexual difficulties who cannot visit a therapist yet have sufficient coping skills and motivation to adhere to the advice and prescriptions included in the self-help method. Self help can facilitate the transfer of treatment gains to the unassisted situation at home and thus reduce generalization difficulties from therapist setting to the home setting. They may prevent the development of client's codependency on therapist support and can help boost the client's sense of self-efficacy. Several self-help approaches were found to be efficacious, both statistically and clinically, in the treatment of several types of sexual dysfunction. The outcome is qualified by problem type, client characteristics, and delivery format of the self-help. The methods that are offered vary from cookbook-like protocols, which need to be followed in a stepwise and strict fashion, to strategies requiring the user to self-adapt a general problem-solving method to his or her personal situation, thereby enabling the individual to become his or her own therapist. PMID- 19308841 TI - "Good sex" and religion: a feminist overview. AB - This article presents an overview of both the processes and the results of an international, interdisciplinary, and interreligious feminist study of "good sex" that resulted in a volume by the same name. We argue that religion (including its secular equivalent, i.e., global capitalism) remains a powerfully influential cultural force that shapes people's lives, in general, and sanctifies their beliefs, in particular, about what makes for good sex. This review seeks to expand conversations about sex in the bedroom and other private arenas (like the confessional) into more public venues and to demonstrate the connections between power, pleasure, and justice. The need to deconstruct religious traditions so as to critically analyze their structures and components is recognized. Several examples of how feminist scholars and activists are retrieving female-friendly religious insights from both their traditions and more transgressive communities of resistance are provided. This article also points to several ways that religious sexual scripts and norms might be reconstructed. Topics addressed include discussions of how to understand footbinding, the tendency of "forbidden" fruit to prove most erotic, whether sexual entanglements are spiritually dangerous distractions, and ways in which religion can make motherhood "compulsory." We examine both the ways in which equating sexual activity with reproductive activity have obscured the value of women's sexual delight and the risks to many women and children of an unqualified validation of sexual pleasure. Both the ambivalence of religious teachings about sexuality and the difficulties posed by monolithic portrayals of religious traditions are identified. PMID- 19308842 TI - An integrative theoretical framework for understanding sexual motivation, arousal, and behavior. AB - An integrative theoretical framework and model for understanding sexual motivation, arousal, and behavior is presented, combining the principles of incentive motivation theory and the hierarchical control of behavior. It is intended to stimulate discussion. The framework can serve as a "route map" in understanding the links between different component processes and their interactions, as well as the relations between different academic perspectives on understanding sexuality. It is suggested that both excitation and inhibition of sexual motivation, arousal, and behavior act at various levels in a hierarchical structure, and much confusion can be avoided by distinguishing these levels. The model integrates information from different branches of psychology: biological, evolutionary, clinical, cognitive, developmental, and social. It describes interactions between sexual behavior and anxiety, attachment, aggression, and drug taking; and it is applied to gender differences, evolutionary psychology, sexual deviancy, sexual addiction, and the biological bases of sexuality. PMID- 19308843 TI - Erotic target location errors: an underappreciated paraphilic dimension. AB - Based on studies of heterosexual male fetishists, transvestites, and transsexuals, Blanchard (1991) proposed the existence of a hitherto unrecognized paraphilic dimension, erotic target location errors (ETLEs), involving the erroneous location of erotic targets in the environment. ETLEs can involve preferential attention to a peripheral or inessential part of an erotic target, manifesting as fetishism, or mislocation of an erotic target in one's own body, manifesting as the desire to impersonate or become a facsimile of the erotic target (e.g., transvestism or transsexualism). Despite its potential clinical and heuristic value, the concept that ETLEs define a paraphilic dimension is underappreciated. This review summarizes the studies leading to the concept of ETLEs and describes how ETLEs are believed to manifest in men whose preferred erotic targets are women, children, men, amputees, plush animals, and real animals. This review also describes ETLEs in women; discusses possible etiologies of ETLEs; considers the implications of the ETLE concept for psychoanalytic theories of transvestism and male-to-female transsexualism, as well as for the forthcoming revision of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition; suggests reasons why the concept of ETLEs has been underappreciated; and describes what might result if the concept were more widely appreciated. PMID- 19308844 TI - The importance of individual differences in pornography use: theoretical perspectives and implications for treating sexual offenders. AB - This article reviews the extant literature regarding pornography's influence on antisocial attitudes, sexual arousal, and sexually aggressive behavior in both noncriminal and criminal samples. The article concludes that when examined in the context of multiple, interacting factors, the findings are highly consistent across experimental and nonexperimental studies and across differing populations in showing that pornography use can be a risk factor for sexually aggressive outcomes, principally for men who are high on other risk factors and who use pornography frequently. Finally, this article presents theoretical implications based on these findings, as well as some clinical implications relevant to the assessment and treatment of sexual offenders. PMID- 19308845 TI - Sexual response in cardiovascular disease. AB - The cardiovascular response to sexual activity in men and women is similar to other daily activities and not excessive between couples in a long-standing relationship. The risk of an ischemic event during sexual activity is low and death very uncommon. Sexual dysfunction is common in cardiac patients and, in men, may occur before cardiac symptoms, with a time interval of 2 to 5 years. Exercise testing fails to identify nonobstructive but vulnerable-to-rupture lipid rich plaques. Cardiac computed tomography identifies these silent plaques and triggers an aggressive risk-reducing management plan. As erectile dysfunction in men over 40 years of age is usually of vascular (endothelial dysfunction) origin, they should be considered "cardiovascular equivalents," and secondary prevention guidelines should be followed. PMID- 19308846 TI - The role of attention in sexual arousal: implications for treatment of sexual dysfunction. AB - The literature on the role of attention in sexual arousal is reviewed, especially that which has implications for noninvasive treatment of sexual dysfunction. Findings suggest that voluntary control of sexual arousal can be achieved through attentional focus on nonsexual cognitions or sexual fantasy. Cognitive biases may direct attention and thus facilitate or impede sexual arousal. Sexual arousal may be influenced by directed attentional focus, and preliminary evidence suggests that mindfulness techniques may result in longer-term changes in attentional focus; these changes, in turn, may improve sexual response. Information processing models of sexual arousal developed in light of such findings are discussed. This research establishes the central role of attentional processes in facilitating physiological and, especially, subjective sexual arousal. Implementing approaches that capitalize on attentional processes could advance noninvasive treatment of sexual dysfunction. Future avenues of research might investigate how play, mammalian play circuits, and flow states are relevant to sexual response and satisfaction. PMID- 19308847 TI - Neurocognitive screening of lead-exposed andean adolescents and young adults. AB - This study was designed to assess the utility of two psychometric tests with putative minimal cultural bias for use in field screening of lead (Pb)-exposed Ecuadorian Andean workers. Specifically, the study evaluated the effectiveness in Pb-exposed adolescents and young adults of a nonverbal reasoning test standardized for younger children, and compared the findings with performance on a test of auditory memory. The Raven Coloured Progressive Matrices (RCPM) was used as a test of nonverbal intelligence, and the Digit Span subtest of the Wechsler IV intelligence scale was used to assess auditory memory/attention. The participants were 35 chronically Pb-exposed Pb-glazing workers, aged 12-21 yr. Blood lead (PbB) levels for the study group ranged from 3 to 86 microg/dl, with 65.7% of the group at and above 10 microg/dl. Zinc protoporphyrin heme ratios (ZPP/heme) ranged from 38 to 380 micromol/mol, with 57.1% of the participants showing abnormal ZPP/heme (>69 micromol/mol). ZPP/heme was significantly correlated with PbB levels, suggesting chronic Pb exposure. Performance on the RCPM was less than average on the U.S., British, and Puerto Rican norms, but average on the Peruvian norms. Significant inverse associations between PbB/ZPP concentrations and RCPM standard scores using the U.S., Puerto Rican, and Peruvian norms were observed, indicating decreasing RCPM test performance with increasing PbB and ZPP levels. RCPM scores were significantly correlated with performance on the Digit Span test for auditory memory. Mean Digit Span scale score was less than average, suggesting auditory memory/attention deficits. In conclusion, both the RCPM and Digit Span tests were found to be effective instruments for field screening of visual-spatial reasoning and auditory memory abilities, respectively, in Pb-exposed Andean adolescents and young adults. PMID- 19308848 TI - Life-cycle exposure of fathead minnows to a mixture of six common pharmaceuticals and triclosan. AB - Fathead minnows were exposed for a life cycle to environmentally relevant concentrations of a mixture of six common pharmaceuticals and one personal care product (nominal concentrations: 1,000, 300, 100, 30, or 10 ng/L). Mean measured concentrations of each chemical in the highest fish exposure aquaria were: naproxen 793 ng/L, gemfibrozil 662 ng/L, diclofenac 331 ng/L, ibuprophen 217 ng/L, triclosan 115 ng/L, salicylic acid 67 ng/L, and acetaminophen (chemical analysis inconclusive, nominal 1000 ng/L). Fish exposed for a life cycle even to the highest concentrations of the six pharmaceuticals and personal care product (PPCP) mixture showed no significant changes in growth and development compared to control. Length, weights, condition factors, liver weights, and gonad weights of PPCP-exposed fish were similar to water and solvent controls (0.000005% ethanol v/v). There were no marked effects of PPCP mixture exposure on external sex characteristics of the fish or on egg production. The only parameter that appeared to be affected was percent larval deformities in F1, which showed a significant increase in the 100- and 300-ng/L (nominal) PPCP mixture. Larvae from control fish had 4.7% (water controls) and 3.4% (solvent controls) deformities, compared to 9.3% in the 100-ng/L (nominal) PPCP mixture and 9.2% deformities in the 300-ng/L (nominal) PPCP mixture. Chronic exposure to environmentally relevant concentrations of seven PPCP most often detected in Canadian municipal wastewater effluents (MWWE) did not appear to affect fathead minnow survival, growth, or egg production, although it produced quantitative increases in deformities in the F1 generation. PMID- 19308849 TI - Oxidative stress due to (R)-styrene oxide exposure and the role of antioxidants in non-Swiss albino (NSA) mice. AB - Styrene produces lung and liver damage that may be related to oxidative stress. The purpose of this study was to investigate the toxicity of (R)-styrene oxide (R SO), the more active enantiomeric metabolite of styrene, and the protective properties of the antioxidants glutathione (GSH), N-acetylcysteine (NAC), and 4 methoxy-L-tyrosinyl-gamma-L-glutamyl-L-cysteinyl-glycine (UPF1) against R-SO induced toxicity in non-Swiss Albino (NSA) mice. UPF1 is a synthetic GSH analog that was shown to have 60 times the ability to scavenge reactive oxygen species (ROS) in comparison to GSH. R-SO toxicity to the lung was measured by elevations in the activity of lactate dehydrogenase (LDH), protein concentration, and number of cells in bronchoalveolar lavage fluid (BALF). Toxicity to the liver was measured by increases in serum sorbitol dehydrogenase (SDH) activity. Antioxidants were not able to decrease the adverse effects of R-SO on lung. However, NAC (200 mg/kg) ip and GSH (600 mg/kg), administered orally prior to R SO (300 mg/kg) ip, showed significant protection against liver toxicity as measured by SDH activity. Unexpectedly, a synthetic GSH analog, UPF1 (0.8 mg/kg), administered intravenously (iv) prior to R-SO, produced a synergistic effect with regard to liver and lung toxicity. Treatment with UPF1 (0.8 mg/kg) iv every other day for 1 wk for preconditioning prior to R-SO ip did not result in any protection against liver and lung toxicity, but rather enhanced the toxicity when administered prior R-SO. The results of the present study demonstrated protection against R-SO toxicity in liver but not lung by the administration of the antioxidants NAC and GSH. PMID- 19308850 TI - Traffic air pollution and lung cancer in females in Taiwan: petrol station density as an indicator of disease development. AB - To investigate the relationship between traffic air pollution exposure and development of lung cancer in females, studies were conducted using a matched cancer case-control model into deaths that occurred in Taiwan from 1997 through 2006. Data on all eligible lung cancer deaths in females were obtained from the Bureau of Vital Statistics of the Taiwan Provincial Department of Health. The control group consisted of women who died from causes other than neoplasms or diseases that were associated with respiratory problems. The controls were pair matched to the cancer cases by year of birth and year of death. Each matched control was selected randomly from the set of possible controls for each case. Data on the number of petrol stations in study municipalities were collected from the two major petroleum supply companies, Chinese Petroleum Corporation (CPC) and Formosa Petrochemical Corporation (FPCC). The petrol station density (per square kilometer; PSD) for study municipalities was used as an indicator of a subject's exposure to benzene and other hydrocarbons present in ambient evaporative losses of petrol or to air emissions from motor vehicles. The subjects were divided into tertiles according to PSD in their residential municipality. The results showed that there was a significant exposure-response relationship between PSD and risk of lung cancer in females after controlling for possible confounders. The findings of this study warrant further investigation of the role of traffic air pollution exposure in the etiology of lung cancer. PMID- 19308851 TI - Occupations and lung cancer: a population-based case-control study in British Columbia. AB - An investigation based on a large population-based case-control study in British Columbia, Canada, was conducted to identify high-risk occupations for lung cancer by histological subtypes. Subjects were 14,755 male incident cancer cases for whom lifetime occupational histories and information on smoking and relevant covariates were collected. Occupational associations for 2998 lung cancer cases, including histological subtypes, were assessed by logistic regression using other cancer cases, excluding smoking-related cancers, as controls. An excess risk of lung cancer was found among workers in metal processing, bakers, and ship deck crew for all histological subtypes, and construction workers, chefs and cooks, and medical workers for specific histological subtypes. Occupational associations that are unique to histological subtypes of lung cancer were identified. Owing to a scarcity of literature in this area, future research needs to focus on confirming these histological associations, and identifying the risk from key exposures found within these occupations (e.g., medical radiation, electromagnetic fields, and cooking fumes). PMID- 19308853 TI - This month in the Scandinavian Journal of Gastroenterology. PMID- 19308854 TI - Interference between outbreaks of epidemic viruses: additional Norwegian observations. PMID- 19308855 TI - Large-volume breast reduction: Long-term results. AB - Forty-three women had reduction mammaplasty during the period 1992-2000 and the cosmetic outcome was evaluated using subjective and objective measures. The inferior pedicle technique was used in all cases. The mean (SD) weight of resected tissue was 1121 (415) g. All objective measurements were within the ideal range except for breast volume and nipple-to-inframammary-line distance, which were more than ideal. Overall, the median difference in measurements between the two breasts of each woman was less than 10%. However, the subjective evaluations given by both clinicians and the patients for overall symmetry and for general aesthetic appearance fell below the preset threshold. The shape of the breast correlated best with the grades of symmetry and general appearance. There was no correlation between the objective and subjective evaluations of symmetry. The disappointing subjective scores, which may be attributed to the specific characteristics of our sample group, are heavy breasts and overweight patients, a double team approach, and the inevitable pseudoptosis that develops with the inferior pedicle approach. These direct us to recommend searching for an alternative operative technique and to evaluate its long term results, as well as placing the nipple-areola complex lower than the standard inframammary fold projection. PMID- 19308856 TI - The nasal alar elevator: a new device that may reduce the need for primary operation of the nose in patients with cleft lip. AB - To improve the shape of the cleft lip nose preoperatively, we have developed the nasal alar elevator. This has been used routinely since 1996 on all our cleft lip patients who have an asymmetrical nose, from the first week after birth until the date of primary lip surgery. We present our 11-year-long experience of using the device on patients born with complete, unilateral cleft lip. In this study 56 children, born between 1996 and 2006 inclusive, with complete unilateral cleft lip, had preoperative treatment with the elevator. During this 11-year period, continuous evaluation during the preoperative period, and its effects on the cleft lip nose, were evaluated, both preoperatively and postoperatively. Our results show that the preoperative use of the device has led to less need for primary nasal surgery. Instead of having to have a primary rhinoplasty (McComb) together with a lip plasty, as a routine, now only about 30% of the patients need primary surgical correction of the nose. If nasal correction is needed, a rather limited undermining of skin over the ala on the cleft side will often be sufficient. The use of a nasal elevator reduces both the length and the extent of the primary intervention, without compromising the final result. PMID- 19308857 TI - Haemangioma of the forehead: radiographic diagnosis and definitive surgical treatment in a Japanese series. AB - Haemangioma of the skull is usually encountered as a solitary lesion, so few separate cases have been reported, and the protocol for treatment has not been confirmed. Here we describe its treatment in five consecutive Japanese patients. Three men and two women aged from 36 to 40 years were studied. The lesions were noticed in the left frontal bone, one in the supraorbital region and the rest in the forehead; they were 1.5-2.5 cm in diameter. The preoperative diagnosis was made precisely by plain and tangential radiographs and computed tomographic (CT) views. Once the lesion was suspected, radical resection and immediate reconstruction were required to prevent recurrence. All five radical resections were reconstructed immediately by split calvarial bone harvested from the ipsilateral parietal bone in the same operative field. The postoperative course was uneventful in all cases, and the postoperative contour of the reconstructed site and the donor site were nearly perfect. There were no particular complications. Two to 10 years have passed without recurrence. PMID- 19308858 TI - Sitting can cause ischaemia in the subcutaneous tissue of the buttocks, which implicates multilayer tissue damage in the development of pressure ulcers. AB - A better understanding of how pressure ulcers develop in the buttocks will improve prophylactic measures. Our aim was to investigate signs of reduced perfusion and ischaemia in the subcutaneous fat in the buttocks during sitting. A microelectrode was used to quantify oxygen (pO(2)). Metabolites that indicate aerobic or anaerobic metabolism (glucose, lactate, pyruvate, and glycerol) were quantified using microdialysis. Sixteen healthy people were studied while they sat on a wheel chair cushion, and a hard surface. Sitting pressures were mapped, and the thickness of the subcutaneous fatty layer was measured. The results showed that pO(2) and glucose were significantly reduced during sitting, and for pO(2) the effect is significantly more profound during sitting on a hard surface. After loading, both glucose and pO(2) increased significantly. We conclude that the subcutaneous adipose tissue covering the ischial tuberosities becomes ischaemic during sitting. This finding supports the theory that not only is the skin involved in early development of pressure ulcers, but also the deeper tissues. PMID- 19308859 TI - Anatomical study of the perforators from the ulnar palmar digital artery of the little finger and clinical uses of digital artery perforator flaps. AB - Our aim was to explore the anatomical features of the cutaneous perforators from the ulnar palmar digital artery of the little finger and to establish the anatomical basis of the ulnar palmar digital artery perforator flap for reconstruction of the ulnar aspect of the palm and fingers. We found that at least one perforator is present between the metacarpophalangeal (MCP) joint and 9 mm proximal to the MCP joint. This finding establishes the anatomical basis of the distal base ulnar palmar digital artery perforator flap for reconstruction of the ulnar aspect of the palm and fingers. PMID- 19308860 TI - Evaluation of a Swedish version of the patient-rated wrist evaluation outcome questionnaire: good responsiveness, validity, and reliability, in 99 patients recovering from a fracture of the distal radius. AB - The patient-rated wrist evaluation (PRWE) form is an established outcome questionnaire designed to measure wrist pain and disabilities in activities of daily living. We translated and validated this score for use in Sweden. The original PRWE score was translated forwards and backwards, and then the responsiveness, validity, and reliability of the Swedish version were tested in 99 patients who were recovering from a fracture of the distal radius. The patients completed the PRWE questionnaire 7 weeks and 4 to 6 months after the injury. Responsiveness was assessed by Standard Response Mean and Effect Size. Content validity was evaluated by examining the proportion of best and worse possible scores. Construct validity was assessed by comparing scores twice after the injury and by comparing patients with fractures of different severity. Criterion validity was evaluated by correlating the PRWE with another upper extremity score: the disability of the arm, shoulder, and hand (DASH) score. Reliability was evaluated with a test-retest and by internal consistency. Responsiveness was excellent (SRM = 1.4-1.7, ES = 1.3). Five patients reported the best possible result after 4 to 6 months but none at 7 weeks, and no patient reported the worst score at any time. The PRWE score corresponded well with improvement and showed good correlation with severity of fracture. The PRWE showed a strong correlation with DASH score (Spearman's rank coefficient = 0.86). Intraobserver reliability of the test-retest and internal consistency was good (Kendall W coefficient = 0.79, Spearman's rank coefficient = 0.99, Cronbach's alpha = 0.94-0.97). We found this Swedish version of the PRWE to be responsive, valid, and reliable for evaluating the patient-rated outcome after a fracture of the distal radius. PMID- 19308861 TI - Modification of the forearm tourniquet techniques of intravenous regional anaesthesia for operations on the distal forearm and hand. AB - Our aim was to find out if a modified intravenous regional anaesthetic block technique, used for invasive surgical procedures on the distal forearm and hand, results in a drier operative field than traditional methods. Twenty consenting adult (age > 18) patients who were to have an operation on the distal forearm or hand were randomised into two groups (n=10 in each). The first group was using a traditional bier block, with a double upper arm tourniquet. The second group was using a modified regional anaesthetic block technique, with a single upper arm tourniquet, and a single forearm tourniquet. All operative fields were recorded photographically and judged by the operating surgeon as "wet" or "dry". Analgesic requirements and subjective pain were recorded. Plasma lignocaine concentrations were measured. "Wet" operative fields were seen in 6 of the conventional and 0 of the modified group (p=0.01). Patients in the modified group were more comfortable during the procedures (p=0.004). This benefit was not sustained postoperatively (p=0.57). Plasma lignocaine concentrations were higher in the conventional group (p=0.004). The modified technique was as safe as the conventional technique but has the benefits of a drier surgical field and improved intraoperative comfort for patients. PMID- 19308862 TI - Atypical Apert syndrome: sequential and segmental distraction osteogenesis of the skull, midface, and mandible. AB - We describe a boy with Apert syndrome, including cranial deformities and syndactyly (acrocephalosyndactyly), though intracranial hypertension, exophthalmos, and midfacial hypoplasia were mild. We treated him by mandibular distraction, in addition to fronto-orbital distraction, and Le Fort III midfacial distraction, with good results. PMID- 19308863 TI - Giant aneurysm of the ulnar artery in the palm treated by resection and microvascular reconstruction. AB - We report a giant aneurysm in the palm that was arising from the ulnar artery. We treated it successfully by resection followed by microvascular reconstruction with a vein graft. PMID- 19308864 TI - Atypical ulnar tunnel syndrome accompanied by sensory disturbance of the dorsal sensory branch of the ulnar nerve. AB - An atypical accessory abductor digiti minimi, which is morphologically a bicipital muscle, was the cause of the ulnar tunnel syndrome. A ganglion grew from the volar carpal joint at the same time and caused sensory disturbance of the dorsal sensory branch of the ulnar nerve. PMID- 19308865 TI - Is image everything? The role of self-image in the relationship between family functioning and substance use among Hispanic adolescents. AB - This cross-sectional self-report study examined (1) whether family functioning (Family Functioning in Adolescence Questionnaire, FFAQ) and self-image (Piers Harris Children's Self-Concept Scale) have independent associations with smoking and alcohol use and (2) whether self-image mediates the relationship between family functioning, smoking, and alcohol use among 1,598 Hispanic males and females in grades 7 through 12 in Los Angeles County, California, in 2001. The findings supported the main effects and, to some degree, the mediational effects that were hypothesized. Limitations and implications for this study are discussed, and directions for future research are suggested. This study was funded by the California Tobacco Related Disease Research Program. PMID- 19308866 TI - Pathways from acculturation stress to substance use among latino adolescents. AB - The purpose of this study was to examine the link between acculturation stress and substance use among Latino adolescents. In-home interviews were completed with the participants at four time-points between 2005 and 2007. Path analysis was completed using longitudinal data from 286 Latino adolescents living in North Carolina and Arizona (65% foreign-born). Results indicate that acculturation stress influences family and friend relationships, which in turn affect adolescent mental health problems, and finally, substance use. Key mediators in the pathway from acculturation stress to substance use were parent-adolescent conflict, internalizing, and externalizing problems. Implications for practice and research have been discussed here. PMID- 19308867 TI - Range data pre-processing for the evaluation of torso shape and symmetry in scoliosis. AB - Pre-processing range scans of the human torso for evaluating shape and symmetry changes in scoliosis are non-trivial. First, stray points from surrounding artefacts are often arbitrarily positioned and not amenable to automatic removal. Second, the asymmetrical alignment of the arms and neck makes cropping them difficult. Third, despite a plethora of methods, removal of holes by surface approximation for this niche application remains a challenge particularly in obscure regions like the sides and armpits. This paper proposes a novel surface approximation method and incorporates it into an integrated procedure for pre processing range scans of the torso that includes interactive tools for cropping stray points and extremities. The new method, spline-fitted moving least squares (MLS), makes use of the Bezier curve and MLS algorithms. Numeric and clinical tests on scans of 30 volunteers, with and without scoliosis, show that the proposed method outperforms its constituent methods and a commercially available graphics package for this application. PMID- 19308868 TI - Knee joint biomechanics in closed-kinetic-chain exercises. AB - Effective management of knee joint disorders demands appropriate rehabilitation programs to restore function while strengthening muscles. Excessive stresses in cartilage/menisci and forces in ligaments should be avoided to not exacerbate joint condition after an injury or reconstruction. Using a validated 3D nonlinear finite element model, detailed biomechanics of the entire joint in closed-kinetic chain squat exercises are investigated at different flexion angles, weights in hands, femur-tibia orientations and coactivity in hamstrings. Predictions are in agreement with results of earlier studies. Estimation of small forces in cruciate ligaments advocates the use of squat exercises at all joint angles and external loads. In contrast, large contact stresses, especially at the patellofemoral joint, that approach cartilage failure threshold in compression suggest avoiding squatting at greater flexion angles, joint moments and weights in hands. Current results are helpful in comprehensive evaluation and design of effective exercise therapies and trainings with minimal risk to various components. PMID- 19308869 TI - An analytical model for the ergometer rowing: inverse multibody dynamics analysis. AB - A model for the ergometer rowing exercise is presented in this paper. From the quantitative observations of a particular trajectory (motion), the model is used to determine the moment of the forces produced by the muscles about each joint. These forces are evaluated according to the continuous system of equations of motion. An inverse dynamics analysis is performed in order to predict the joint torques developed by the muscles during the execution of the task. An elementary multibody mechanical system is used as an example to discuss the assumptions and procedures adopted. PMID- 19308870 TI - A semi-automated method for hexahedral mesh construction of human vertebrae from CT scans. AB - Generation of finite element (FE) meshes of vertebrae from computed tomography (CT) scans is labour intensive due to their geometric complexity. As such, techniques that simplify creation of meshes of vertebrae are needed to make FE analysis feasible for large studies and clinical applications. Techniques to obtain a geometric representation of bone contours from CT scans of vertebrae and construct a hexahedral mesh from the contours were developed. An automated edge detection technique was developed to identify surface contours of the vertebrae, followed by atlas based B-spline curve fitting to construct curves from the edge points. The method was automatic and robust to missing data, with a controllable degree of smoothing and interpolation. Parametric mapping was then used to generate nodes for each CT slice, which were connected between slices to obtain a hexahedral mesh. This method could be adapted for modelling a variety of orthopaedic structures. PMID- 19308871 TI - The use of artificial neural networks to reduce data collection demands in determining spine loading: a laboratory based analysis. AB - The extensive data requirements of three-dimensional inverse dynamics and joint modelling to estimate spinal loading prevent the implementation of these models in industry and may hinder development of advanced injury prevention standards. This work examines the potential of feed forward artificial neural networks (ANNs) as a data reduction approach and compared predictions to rigid link and EMG-assisted models. Ten males and ten females performed dynamic lifts, all approaches were applied and comparisons of predicted joint moments and joint forces were evaluated. While the ANN under- predicted peak extension moments (p = 0.0261) and joint compression (p < 0.0001), predictions of cumulative extension moments (p = 0.8293) and cumulative joint compression (p = 0.9557) were not different. Therefore, the ANNs proposed may be used to obtain estimates of cumulative exposure variables with reduced input demands; however they should not be applied to determine peak demands of a worker's exposure. PMID- 19308872 TI - The relationship between heart rate intensity and pedometer step counts in adolescents. AB - The primary objective of this study was to examine the relationship between heart rate intensity and pedometer step counts in adolescents. To determine cardiorespiratory fitness, 106 participants (47 boys, 59 girls, mean age 14.2 years, s = 0.8) completed the Queen's College Step Test and were classified as having low, moderate or high cardiorespiratory fitness. Adolescents also completed a 10-min treadmill trial while wearing a pedometer and heart rate monitor. The participants were instructed to maintain their heart rate between 65 and 75% of their maximum heart rate while running or walking on a treadmill. A heart rate of 65-75% maximum was associated with 146 steps per minute (s = 22) in boys and 137 steps per minute (s = 22) in girls. Results of analysis of variance indicated that there was a main effect for level of fitness (F(2,102) = 9.36, P < 0.001). The correlation between mean steps per minute and estimated maximum oxygen consumption was statistically significant (r = 0.44, P < 0.001). The results from this study suggest that a step rate of 130 steps per minute is equal to 65-75% maximum heart rate in low-fit adolescents and achieving 130 steps per minute could be used as an initial goal to improve fitness. PMID- 19308873 TI - Peak athletic performance and ageing: evidence from baseball. AB - Baseball players exhibit a pattern of improvement and decline in performance; however, differing lengths of careers and changes in rules and characteristics of the game complicate assessments of age-related effects on performance. This study attempts to isolate the impact of age on several player skills while controlling for relevant outside factors using longitudinal data from 86 seasons of Major League Baseball. The results indicate that players age in different skills in accord with studies of ageing in other athletic contests. For overall performance, multiple-regression estimates indicate that hitters and pitchers peak around the age of 29 - later than previous estimates. Athletic skills such as hitting and running peak earlier than skills that rely heavily on experience and knowledge, such as issuing and drawing walks. PMID- 19308874 TI - Interpretation and application of the offside law by expert assistant referees: perception of spatial positions in complex dynamic events on and off the field. AB - Baldo et al. (2002) and Helsen et al. (2006) considered the flash-lag effect to explain errors made by assistant referees when judging offside in association football. The main aims of the present study were as follows: (1) to determine whether the flash-lag effect emerges in offside situations on the field of play or off the field when presented as computer animations or as video footage of real-life matches; (2) to examine offside decision-making errors in two standards of assistant referee - international FIFA and Belgian national referees. The results support the flash-lag hypothesis in several ways. First, both the FIFA and Belgian assistant referees were more likely to make errors by raising their flag when they had to assess offside situations on the field of play and when presented as three-a-side computer animations. Second, more flag errors were made when the defender moved in the opposite direction to that of the attacker. Third, the strategy of raising the flag in case of doubt was not observed when an interpretation of the offside law had to be made about the involvement of play of an attacker. Future research is needed to examine the extent to which on- and off the-field training sessions can be used as training tools to improve offside decision-making. PMID- 19308875 TI - Enzyme activity and myoglobin concentration in rat myocardium and skeletal muscles after passive intermittent simulated altitude exposure. AB - We studied the effect of intermittent hypobaric hypoxia exposure on lactate dehydrogenase and citrate synthase activities, together with myoglobin content, of rat myocardium, tibialis anterior, and diaphragm muscles. The intermittent hypoxia exposure programme consisted of daily 4-h sessions in a hypobaric chamber (5000 m) over a period of 22 days. Samples were taken at the end of the programme, and 20 and 40 days later, and compared with those of control animals. In myocardium, lactate dehydrogenase activity was significantly depressed in animals 20 days post-exposure (314.6 +/- 15.3 IU . g(-1)) compared with control animals (400 +/- 14.3 IU . g(-1)), while citrate synthase activity and myoglobin concentration showed a significant stepwise increase from control animals (88.2 +/- 3.6 IU . g(-1) and 4.38 +/- 0.13 microm . mg(-1)) to animals 20 days (104.7 +/- 3.7 IU . g(-1) and 5.01 +/- 0.17 microm . mg(-1)) and 40 days post-exposure (108.8 +/- 6.5 IU . g(-1) and 5.11 +/- 0.22 microm . mg(-1)). In contrast, no differences were found in diaphragm and tibialis anterior muscles. Our results show that intermittent hypobaric hypoxia exposure increased the oxidative character of myocardium even 20 days after the hypoxic stimulus has ceased, and that this effect lasts for more than 40 days for citrate synthase activity and myoglobin concentration. These findings support our previous results on skeletal and cardiac muscle capillarization after passive intermittent simulated altitude exposure, thus providing morphofunctional and biochemical evidence for increased cardiac aerobic efficiency. PMID- 19308876 TI - Functional bowel symptoms and GnRH antibodies: common findings in patients with primary Sjogren's syndrome but not in systemic sclerosis. PMID- 19308877 TI - Distinct haplogenotypes of the dopamine D2 receptor gene are associated with non smoking behaviour and daily cigarette consumption. AB - INTRODUCTION: Dopamine systems in the CNS are decisively implicated in the motivational and rewarding properties of nicotine. The dopamine D2 receptor (DRD2) plays a pivotal role by promoting these properties, making this gene a good candidate for association studies. Several single nucleotide polymorphisms (SNPs) have been described to influence the expression of DRD2. The amount of expressed DRD2 will finally be the result of the sum and/or interaction of several functional polymorphisms located at the respective DNA strand forming a distinct haplotype. Thus, the knowledge about the distribution of the haplotypes in groups of subjects, differing by their smoking behaviour, would result in a better understanding of the putative associations compared to single SNP investigations. METHODS: 218 healthy subjects grouped for being never smokers, former smokers, and current smokers, were genotyped for the following polymorphisms: -141 ins(I)/del(D), STRPi2 (intron 2), C957T (exon 7), A1385G (exon 8), and TaqIA. Regular immoderate alcohol consumption was an exclusion criterion. RESULTS: In the total study group four haplotypes represented 90% of the haplotypes, with I-T-A-A2, I-C-G-A2, I-C-A-A1, and D-C-G-A2 accounting for around 50%, 20%, 10%, and 10%, respectively. I-C-G-A2 homozygosity was significantly higher in never smokers compared to ever smokers (current+former smokers) (chi2=36.585, df=1, p<0.001). There was a significant difference in the daily cigarette consumption of current smokers with respect to the haplogenotype (chi2=3211.9, df=18, p=0.003). Current smokers with a haplogenotype containing at least one I-T-A-A2 allele showed a significant smaller daily cigarette consumption (15.1+/-7.93) compared to subjects with a genotype not bearing this allele (20.1+/-6.79; T=-2.06, df=61, p=0.044). CONCLUSION: We have demonstrated an association of the distinct haplogenotype I/I-C/C-G/G-A2/A2 of the DRD2 gene with a reduced risk to become a smoker in Caucasians of German origin. This protection may result from an association of this haplotype with a reduced activation of the dopaminergic neurotransmission by nicotine. Moreover, in current smokers, higher daily cigarette consumption is associated with those haplogenotypes that do not contain the I-T-A-A2 haplotype. PMID- 19308878 TI - The prognostic impact of psychotropic drugs in intentional drug overdose. AB - INTRODUCTION: Psychotropic drugs belong to the drugs most frequently involved in intentional drug overdose (IDO). Few studies have explored their prognostic impact during hospitalisation for IDO. METHODS: In order to assess which types of psychotropic drugs ingested during IDOs were associated with an increased morbidity, a cohort study included 1,974 patients consecutively hospitalised for IDO. IDOs were categorised as serious if associated with one of the following criteria: death, hospitalisation longer than 48 h, respiratory support, vasopressive drugs, cardiac massage or dialysis. RESULTS: Nearly all the patients ingested psychotropic medications during the IDO (88.4%), most often benzodiazepines (71.6%). Serious IDO was associated with tricyclics (OR 5.7; 95% CI 3.3-9.8), lithium (OR 4.3; 95% CI 1.6-11.6), carbamates (OR 2.7; 95% CI 1.8 4), anticonvulsants (OR 2.4: 95% CI 1.4-4.3), first-generation antipsychotics (OR 2.4; 95% CI 1.7-3.5) or selective serotonin reuptake inhibitors (SSRIs) (OR 1.6; 95% CI 1.1-2.3). DISCUSSION: Some drugs may be dangerous because of low toxic doses; hence, prescriptions of short duration may be recommended. Moreover, for safety reasons, prescribers may prefer SSRIs to tricyclics and benzodiazepines to carbamates or phenothiazines. PMID- 19308879 TI - Relationship between neuroleptic extrapyramidal syndromes and patients' all-cause mortality. AB - INTRODUCTION: It is important to understand factors contributing to a neuroleptic related increased mortality risk. The objective of this study was to test whether the occurrence of neuroleptic-induced extrapyramidal syndromes (EPS) including tardive dyskinesia (TD) is associated with an increased patients' all-cause mortality. METHODS: In 1995, a sample of 200 patients on neuroleptics was assessed with regard to the presence of Parkinson syndrome, akathisia, and TD. By 2003-2004, i.e., during the following 8-9 year period, 63 patients had died. Patients who had died were compared with 120 patients known to be still alive with regard to several socio-demographic variables and the presence of EPS at the first examination. RESULTS: At the basic assessment, there were no significant differences between patients later still alive and deceased patients with regard to TD. The deceased patients were more frequently women, older, suffered more frequently from an organic disorder, had higher average scores for Parkinson syndrome and less frequently akathisia. Multivariate analysis confirmed age as the only factor contributing to the group difference. Repeating the meta-analysis by Ballesteros et al. (2000) after inclusion of our data, TD remains a weak but a significant predictor of death (OR=1.4). DISCUSSION: Neuroleptic-induced EPS of parkinsonism, akathisia, and TD did not contribute to the patients' all-cause mortality in this study. The association between TD and mortality merits further attention. PMID- 19308880 TI - Confusion, a rather serious adverse drug reaction with valproic acid: a review of the French Pharmacovigilance database. AB - INTRODUCTION: Confusion is an adverse drug reaction frequently observed with valproic acid. Some case reports are published in the literature but no systematic study from a sample of patients has been published. We performed this study in order to describe the main characteristics of this adverse drug reaction. METHODS: Using the French Pharmacovigilance database, we selected the cases of confusion reported since 1985 with valproic acid. RESULTS: 272 cases of confusion were reported with valproic acid: 153 women and 119 men. Confusion mostly occurred during the two first weeks following valproic acid exposure (39.7%). It was "serious" for almost 2/3 of the patients (62.5%) and its outcome favourable in most of the cases (82%). The occurrence of this ADR was more frequent in patients aged between 61 and 80 years. CONCLUSION: This work shows that confusion with valproic acid is a serious, rather frequent but reversible adverse drug reaction. It occurs especially in older patients and during the first two weeks of treatment. PMID- 19308881 TI - Drug prescription patterns in schizophrenia outpatients: analysis of data from a German health insurance fund. AB - INTRODUCTION: The aim of this study was to investigate routine administrative data from a major German health insurance fund, Techniker Krankenkasse, which covers 5.4 million insured individuals. Using a retrospective cohort design, this study analysed data collected from patients with a hospital diagnosis of schizophrenia in 2003 (index hospitalisation) in order to evaluate prescription patterns of antipsychotic drugs. METHODS: Patients with an ICD-10 diagnosis of schizophrenia, at least one year prior membership with the insurance fund and a follow-up period of one year were identified. Results were standardised by age and stratified by the severity of their illness, defined by the number of hospital bed days during the three years preceding the index hospitalisation. RESULTS: A total of 3,121 patients with schizophrenia (male 56.4%, female 43.6%) received 56 692 single prescriptions of antipsychotics. Of these, 35.4% of the prescriptions were for typical and 64.6% for atypical antipsychotics; 55% were for high-potency, 45% for low-potency typical antipsychotics. The most frequently prescribed drugs were olanzapine (26.6%), clozapine (21.3%) and risperidone (19%). There were no relevant gender differences concerning prescription patterns. During a 12-month follow-up period after the first hospitalisation, 1 372 patients (43.9%) were treated exclusively with an atypical antipsychotic, another 499 patients (16%) had a combination of an atypical plus a low-potency typical antipsychotic. Thus, basal therapy with an atypical was observed in 59.9% of our study population. Only 327 patients (10.5%) were treated exclusively with a typical antipsychotic. A total of 645 patients (20.7%) were treated with a combination of atypical plus typical antipsychotic. Changes of medication within one substance group occurred more often with typical antipsychotics (50%) as compared to atypical antipsychotics (25%). DISCUSSION: At 60%, the proportion of patients in this study treated with atypical antipsychotics was surprisingly high. Of significant interest is the frequent prescription of clozapine (14%). The results are discussed in comparison to comparable studies from other countries. PMID- 19308882 TI - Suicidal risks during treatment of bipolar disorder patients with lithium versus anticonvulsants. PMID- 19308883 TI - Metabolic considerations in a case of pregabalin-induced edema. PMID- 19308884 TI - Severe protracted alcohol withdrawal syndrome: prevalence and pharmacological treatment at an inpatient detoxification unit--a naturalistic study. PMID- 19308885 TI - Paliperidone in an adult patient with Asperger syndrome: case report. PMID- 19308886 TI - Cardiac left bundle branch block and pancytopenia in anorexia nervosa: higher risk with mirtazapine and pantoprazole? Case report. PMID- 19308888 TI - Laboratory diagnostics and therapy in thrombosis and hemostasis: from bedside to bench to bedside. PMID- 19308889 TI - Milestones and perspectives in coagulation and hemostasis. AB - Hemostasis is traditionally defined as the physiologic process whereby bleeding is antagonized and possibly stopped to minimize blood loss. The first medical description of the clinical and inherited features of hemostasis can be dated back more than 1000 years, when Abu al-Qasim Khalaf ibn 'Abbas al-Andalusi al Zahrawi' medical treatise provided some initial insights into this puzzling process. Since then, continuous and revolutionary scientific developments have contributed to decoding several aspects of this intricate but essential physiologic phenomenon, providing a reliable model to explain the leading mechanisms involved. Although the point at which bleeding stops is commonly referred to as "coagulation," blood coagulation is actually only one part of a two-part hemostatic process that develops through sequential steps referred to as primary and secondary hemostasis. Throughout its activation and development, the coagulation cascade is strictly regulated by a series of natural inhibitors, which prevent unnecessary and excessive clotting. The aim of this article is to provide a concise overview of the major discoveries and past and current perspectives in coagulation and hemostasis. PMID- 19308890 TI - Circadian variation within hemostasis: an underrecognized link between biology and disease? AB - Biological rhythms are a universal phenomenon in living organisms and serve to help organisms adapt within a circadian cycle to the 24-hour-oscillating environment. The rhythmic modulation of selective pathways thus enables organisms to optimize their ability to store and generate chemical energy, to minimize environmental stresses, and to reproduce by cycles of cell growth and division. Remarkably, the onset of both cardiovascular disorders and venous thromboembolism also undergoes circadian oscillations, which might be closely related to an internal biological clock. A highly repetitive rhythmic cycle seems to modulate platelet and endothelial functions, as well as the concentration and activity of several proteins of the coagulation and fibrinolytic systems. Although it is currently unknown how the global hemostatic efficiency might be affected by these circadian variations, understanding the nature, clinical significance, and pathophysiologic consequences of our hemostatic clock may be helpful for the prevention and treatment of a variety of conditions where either the severity of the illness or therapeutic efficacy exhibit circadian rhythmicity. In this review, clinical and laboratory data describing the effects of these cyclical rhythms on the hemostatic system are discussed. PMID- 19308891 TI - Laboratory monitoring of anticoagulation: where do we stand? AB - The treatment of choice for acute venous thromboembolism is anticoagulant therapy with fast-acting drugs (unfractionated or low-molecular-weight heparin or fondaparinux) aimed at preventing thrombus extension, followed by extended prophylaxis with vitamin K antagonists aimed at preventing recurrence. Experience accumulated over the years has demonstrated that strict laboratory monitoring is required for unfractionated heparin and vitamin K antagonists, making use of these drugs problematic for patients and physicians and prompting researchers to develop new anticoagulants equally effective but without the requirement for laboratory monitoring. The results of clinical trials to date, albeit limited, suggest that these new drugs will probably keep their promise. However, the definitive answer will come subsequent to these clinical trials, when clinicians will start to use these drugs to treat patients in the real world. It is likely that some sort of laboratory monitoring will be required at least for selected categories of patients. Accordingly, clinical laboratories should still be prepared to monitor patients, although the numbers may hopefully decrease sharply in the next decade or so. PMID- 19308892 TI - Pharmacogenetics in hemostasis: friend or foe? AB - Pharmacologic therapies are essential in the management of patients with hemostatic and thrombotic diseases because these therapies are able to modify components of the coagulation pathway and platelet response. Nevertheless, responses to different drugs vary significantly, and the best clinical outcome frequently involves a delicate risk/benefit balance. The recent exponential growth of pharmacogenomics has led to the emergence of individualized medicine that has revolutionized modern medical practice, allowing for a deeper understanding of pathophysiology, increased diagnostic specificity, and better markers for risk stratification and an enhanced potential for gene therapy. Management of drugs prescribed to treat thrombotic and hemostatic abnormalities may benefit from pharmacogenetics, and our focus in this review will be on the pharmacogenetics related to some of the more common drugs that fall into this category. PMID- 19308893 TI - The role of D-dimer testing in patients with suspected venous thromboembolism. AB - D-dimer, the final degradation product of cross-linked fibrin, is typically elevated in patients with acute venous thromboembolism. With its high sensitivity and negative predictive value, D-dimer testing may have a role for ruling-out the diagnosis in patients with suspected deep vein thrombosis or pulmonary embolism. For this purpose, D-dimer testing has been integrated in sequential diagnostic strategies including those using pretest clinical probability assessment and imaging techniques. A large variety of assays are now available for D-dimer measurement, with different sensitivities and specificities for the diagnosis of venous thromboembolism. Attempts to standardize the various D-dimer assays have been made but without any definitive answers as yet. The diagnostic yield of D dimer testing is affected not only by the choice of the appropriate assay but also by patient characteristics. As a consequence, the clinical usefulness of D dimer testing for the exclusion of suspected venous thromboembolism should be carefully evaluated in special clinical settings. PMID- 19308894 TI - Toward a new paradigm for the identification and functional characterization of von Willebrand disease. AB - The diagnosis and functional characterization of von Willebrand disease (VWD) is challenging. There are inherent difficulties in both its identification and classification because of clinical uncertainty, the limitations in the test processes and test panels typically used by laboratories, and because the classification scheme does not always allow unequivocal assignment of any subtype. This article reviews current thoughts and alternatives to the classic approach of the classification and functional characterization of VWD. Of particular interest to this author is the utility of an extended core test panel that includes additional functional VWF assays, such as the collagen binding assay, and the potential for desmopressin (DDAVP) challenge to not only provide therapeutic information but also assist in the better characterization of individuals with defects or deficiencies in von Willebrand factor (VWF). The potential use of supplementary assays such as the PFA-100 and the VWF propeptide assay after DDAVP challenge is also worth noting. PMID- 19308895 TI - Extracorporeal immunoadsorption for the treatment of coagulation inhibitors. AB - Extracorporeal immunoadsorption is a widely used technique for the removal of pathogenic antibodies in a variety of immunologic disorders. This procedure has been used in patients with high-titer inhibitors against coagulation factors for the temporary removal of antibodies before initiating replacement therapy to achieve hemostasis and stop acute bleeding or to cover a surgical procedure. Inhibitor removal by immunoadsorption has also been included at the onset of immune tolerance protocols in both acquired and congenital hemophilia. This article summarizes the current knowledge on the use of this technique in patients with inhibitors against coagulation factors. Overall, the published literature documents that extracorporeal immunoadsorption is a safe and useful technique for the elimination of coagulation inhibitors. However, further randomized clinical trials are needed to better assess the cost-effectiveness of such procedures. PMID- 19308896 TI - Gene therapy for hemophilia: clinical trials and technical tribulations. AB - As monogenic disorders, hemophilia A and B are compelling candidates for treatment with gene therapy. In hemophilia, a therapeutic benefit achieved by gene therapy should only require a modest increase in the endogenous factor level; response to treatment can be monitored easily; and there are relevant small and large animal models. The two main approaches aiming to restore factor VIII or factor IX production are based on genetically modified cells or direct in vivo gene delivery using viral or plasmid vectors. The progress toward gene therapy for hemophilia A and B in both preclinical and clinical models will be evaluated in this review. Various viral and nonviral vectors are discussed in the context of current hurdles arising from preclinical and clinical trials. Despite disappointing clinical results to date, there are favorable indications that the near future should deliver on the long-sought promise of a cure for hemophilia. PMID- 19308897 TI - Procoagulant snake toxins: laboratory studies, diagnosis, and understanding snakebite coagulopathy. AB - Procoagulant toxins are important hemotoxins that have been investigated both as laboratory reagents and potential therapeutic agents. In human envenomation by some elapid and many viperid snakes, these toxins result in venom-induced consumption coagulopathy. Overall, the coagulant activity of the various venoms is difficult to characterize, and many studies simply characterize toxin conversion of isolated substrates, such as the effect of a snake toxin on purified fibrinogen, or on multiple single substrates. As the full effects of toxins on the coagulation pathway are rarely examined, even in vitro, our understanding of the pathophysiology of envenoming is limited. Although prothrombin activators cause a single effect in vitro, there may be complete consumption of fibrinogen, factor V, and factor VIII in vivo due to the downstream effects of the thrombin that is formed. Laboratory diagnosis is a key part of the treatment of snakebite coagulopathy. Assessing which assays are the most informative in snake envenoming, based on the pathophysiology of snakebite coagulopathy, will optimize diagnosis and timing of appropriate coagulation tests. A better understanding of the coagulation effects arising from human envenoming will also improve treatment with antivenom and define the role of adjuvant therapies such as factor replacement. PMID- 19308898 TI - Assessment of thrombin generation: useful or hype? AB - Thrombin plays an important role in hemostasis through its multiple functions across blood coagulation, platelet activation, and fibrinolysis. The measurement of thrombin generation is therefore viewed as a potentially useful test that could be applied to the screening, monitoring, and/or diagnosis of hemostatic abnormalities. Indeed, advances in thrombin generation assays have created significant interest and debate as to whether they may provide a more physiologically relevant testing system than do traditional coagulation tests. A variety of thrombin generation assays, including commercially available systems, have been investigated for their correlation with hypocoagulable and hypercoagulable states. Although there is an extensive body of literature that has investigated the application of thrombin generation assays, some limitations remain. These include poor standardization of reagents and methods and a lack of large prospective studies that demonstrate clear relationships between thrombin generation with bleeding and thrombosis phenotypes, as well as with monitoring anticoagulation. Whether thrombin generation assays become more "useful" than "hype" will require well-designed, large, prospective multicenter trials using standardized methods. PMID- 19308899 TI - Glycemic index, glycemic load, and thrombogenesis. AB - Hyperglycemia and insulin resistance are independent risk factors for cardiovascular disease (CVD). Postprandial glycemic "spikes" adversely affect vascular structure and function via multiple mechanisms including oxidative stress, inflammation, low-density lipoprotein oxidation, protein glycation, and procoagulant activity. Glycemic responses can be reliably predicted by considering both the quantity and quality of carbohydrate. The glycemic index (GI), a measure of carbohydrate quality, has provided insights that knowledge of the sugar or starch content has not. In prospective observational studies, dietary GI and/or glycemic load (GL; the product of the amount of carbohydrate and GI) independently predict CVD, with relative risk ratios of 1.2 to 1.9 comparing highest and lowest quartiles. In randomized controlled trials in overweight subjects, diets based on low GI carbohydrates have decreased plasminogen activator inhibitor-1 activity and other CVD risk factors over and above that of conventional low-fat diets. Taken together, the findings suggest that clinicians may be able to improve CVD outcomes by recommending the judicious use of low GI/GL foods. PMID- 19308900 TI - Biological variation in tests of hemostasis. AB - The two components of biological variability are interindividual variability, which is the variability due to the heterogeneity of physiologic influences among subjects, and intraindividual variability, which is due to the variability in the same individual over time. Analysis of biological variation is crucial for estimating the critical difference, which corresponds with a threshold suggestive of a statistically significant difference between two consecutive results of a laboratory parameter in the same subject and is therefore unlikely attributable to casual (random) oscillation of values. Studies on biological variation of tests of hemostasis are outdated, and the published results should be confirmed by using modern, fully automated methods. Biological variation for coagulation screening tests (prothrombin time and activated partial thromboplastin time) is low and comparable with the values registered for hematologic parameters. However, the index of individuality (ratio between intraindividual and interindividual variability) suggests that the usual preoperative screening for coagulation disorders is influenced by the between-subjects variability. Thrombin time is very constant within and between subjects. Proteins such as fibrinogen, clotting factors, and antithrombin show a low biological variability. In contrast, fibrinolytic parameters, such as plasminogen activator inhibitor 1 and fibrinopeptide A, show very high variability, and their interpretation in the clinical setting must take this into consideration. PMID- 19308903 TI - [Reducing the burden of chronic respiratory diseases]. PMID- 19308901 TI - [Spectrum and bleeding complications of sonographically guided interventions of the liver and pancreas]. AB - PURPOSE: Sonographically guided fine-needle punctures (p.) are considered to be a low risk procedure. Interventions with needles with a larger diameter seem to cause more complications. In search of potential complications, we examined 1923 sonographically guided interventions of the liver and pancreas in a retrospective analysis. MATERIALS AND METHODS: We examined the coherence of the kind of intervention and complications. We considered bleeding with a need for transfusion and/or a need for surgical treatment as complications. Diseases and medication increasing the probability of post-interventional bleeding were also detected. RESULTS: 1923 sonographically guided interventions in the abdomen (1800 in the liver, 123 in the pancreas) were analyzed (n = 1696 diagnostic interventions, n = 227 therapeutic interventions). Needles with diameters > 1 mm were primarily used. Drainage and radiofrequency ablation (RFA) (12 % of all interventions) were performed with devices with diameters between 2 - 3.96 mm. A need for transfusion was found in 8 / 1923 patients (0.4 %), predominantly in the first 24 h. There was no significant correlation between coagulation preventing drugs (heparin, NSAIDs, antiaggregants) and bleeding events. Patients who suffered from liver cirrhosis with a thromboplastin time of < 50 % had a higher risk of post-interventional bleeding than patients with liver cirrhosis and a thromboplastin time > 50 %. Furthermore, therapeutic interventions showed higher complication rates than diagnostic procedures. CONCLUSION: Severe bleeding complications with a need for transfusion in sonographically guided procedures are rare (0.4 %). Our results showed that liver cirrhosis with a low thromboplastin time (< 50 %) seems to be the most important risk factor for patients. Overall, sonographically guided interventions are safe and have low complication rates considering careful performance and contraindications. PMID- 19308904 TI - [Association of A/T polymorphism of the CHRM2 gene with bronchodilator response to ipratropium bromide in asthmatic children]. AB - INTRODUCTION: The aim of this study was to analyze the possible association of A/T polymorphism of the CHRM2 gene with asthma, and pharmacogenetic analysis of the polymorphism with bronchodilator response to ipratropium bromide, an anticholinergic drug used in asthma. MATERIAL AND METHODS: Analysis was performed in a group of 113 children diagnosed with bronchial asthma, and in a group of 123 healthy children from a control group. Moreover, in the group of 32 asthmatic children without concurrent treatment with long-acting beta2-agonists, bronchodilator response to ipratropium bromide was evaluated by the spirometric lung function test. Genetic analysis was performed for A/T polymorphism (rs6962027) of the CHRM2 gene. Genotyping was done with the PCR-RFLP method. Statistical analysis was performed using Statistica v.7.1 software. RESULTS: No association of A/T polymorphism was found with asthma (p=0.865 for genotypes and p=0.782 for alleles). In the pharmacogenetic analysis, it was observed that patients carrying TT genotype of CHRM2 gene polymorphism demonstrated significantly poorer response to anticholinergic drug as compared to the patients with other genotypes for this polymorphism (p=0.035). CONCLUSIONS: We found that TT genotype in the CHRM2 gene was associated with poor bronchodilator response in asthmatic patients. The results should be analyzed carefully considering the small sample size and should be confirmed by other research groups. PMID- 19308905 TI - [Treatment outcomes in culture positive pulmonary tuberculosis]. AB - INTRODUCTION: The aim of the study was to evaluate treatment outcomes in sputum culture positive patients with tuberculosis in three Polish provinces (Warsaw, Gdansk and Siedlce Provinces) in 1995 and again in 2000. We also assessed whether the implementation of the Directly Observed Therapy Short Course (DOTS) strategy in the former Gdansk Province led to improved outcomes compared to the outcomes observed in 1995, when this strategy was not being followed. MATERIAL AND METHODS: We started the study by reviewing microbiology registers covering the years 1995 and 2000 from all the tuberculosis laboratories in three provinces (Warsaw, Gdansk and Siedlce Provinces) and identified sputum culture positive patients. We then reviewed inpatient and outpatient medical records of patients who had been with pulmonary tuberculosis diagnosed and confirmed by bacteriology in 1995 and 2000. Treatment outcomes were evaluated in accordance with the World Health Organisation (WHO) recommendations and classified as: cure, treatment completed, default, treatment failure, death or other. RESULTS: A total of 708 patients were included in the study: 373 diagnosed in 1995 and 335 diagnosed in 2000. According to the WHO criteria, the treatment success rate (the sum of cures and treatment completions) in 1995 and 2000 was 58.8% and 54.0%, the default rate was 15.5% and 17.9%, the failure rate was 2.4% and 2.7% and the death rate was 5.6% and 6.3%, respectively. The rate of outcomes classified as "other" was 18.2% and 22.1%, respectively. Following the implementation of the DOTS strategy in Gdansk Province, treatment outcomes in significantly improved in 2000 compared to the year 1995. The treatment success rate was 89.6% vs 69.3% (p = 0.0037), the default rate was 0.0% vs. 14.7% (p = 0.0005) and the death rate was 0.0% vs. 9.3% (p = 0.0184). CONCLUSIONS: The treatment success rate (the rate of cures and treatment completers) in 1995 and 2000 was 58.8% and 54.0% and was lower than that recommended by WHO (at least 85%). The results demonstrated that the treatment outcomes in the former Gdansk Province in 2000, following the implementation of the DOTS strategy, were significantly better than those in 1995, when the strategy was not being followed. Treatment success was observed in 89.6% and 69.3% of the patients, respectively (p = 0.0037). PMID- 19308906 TI - [Abnormalities in high-resolution computed tomography of the lungs in patients with idiopathic pulmonary arterial hypertension -- correlation with hemodynamic parameters and prognostic significance]. AB - INTRODUCTION: The risk stratification in idiopathic pulmonary arterial hypertension (IPAH) patients is currently based on haemodynamic and functional parameters as well as serum biomarker concentrations. Until now the importance of changes appearing in high-resolution computed tomography (HRCT) of the lungs of patients with IPAH has not been investigated. MATERIAL AND METHODS: Lung HRCT scans were analysed retrospectively in 48 IPAH patients (patients): 37 women, 11 men, mean age 41 +/- 15 years. RESULTS: Focal ground-glass opacifications (FGG) were found in 12 patients (25%), and centrilobular nodules (CN) were found in 8 patients (17%). In the remaining 58% of patients HRCT revealed no changes (N). Significantly lower stroke volume was found in the CN group (41.0 +/- 8.5 ml) compared to 60.8 +/- 15.1 ml in the FGG group and 58.1 +/- 18.0 ml in the N group (p = 0.03). Right atrial pressure was significantly higher in the CN group (12.2 +/- 4.86 mm Hg) than in the FGG group (6.9 +/- 3.9 mm Hg) and the N group (7.6 +/ 5.3 mm Hg), p = 0.047. The presence of nodules was combined with considerably increased risk of death, both in univariate analysis (HR 5.35, 95% CI: 1.16-24.7, p = 0.03) and in multivariate analysis (HR 6.98, 95% CI: 1.41-34.59, p = 0.02). Ground-glass opacifications correlated neither with haemodynamic nor functional indexes, and were of no prognostic significance. CONCLUSIONS: The presence of centrilobular nodules in lung HRCT scans of IPAH patients was combined with more severe haemodynamic compromise and was an independent negative prognostic indicator. PMID- 19308907 TI - [Style of coping with stress and emotional functioning in patients with asthma]. AB - INTRODUCTION: The evaluation of styles of coping with stress resulting from a somatic disease was always considered controversial, since the stress is a consequence of the complexity of the disease as a stressful event and is linked with every field of human life and activity. The main aim of this study was to evaluate the effectiveness of various styles of coping with stress - task oriented, emotion-oriented, or avoidance-oriented - used by patients with asthma to cope with their disease and its consequences. MATERIAL AND METHODS: 100 patients with asthma were examined including 15 with mild asthma, 40 with moderate asthma, and 45 with severe asthma. The first group consisted of patients with mild-to-moderate asthma and the second group of patients with severe asthma. The subjects were mainly females (69% women, 31% men) aged from 19 to 75 years, with at least primary level of education. The following questionnaires were used: Coping Inventory for Stressful Situations (CISS), UWIST Mood Adjective Check List (UMACL), and Asthma Control Test (ACT). RESULTS: Task-oriented style had a positive influence on emotions (p < 0.05) in patients with moderate asthma. Avoidance-oriented style had a positive influence on emotions (p < 0.05) in patients with severe asthma. Emotion-oriented style proved ineffective and had a negative influence on emotions (p < 0.05) in all patients. CONCLUSIONS: Coping style plays a key role in the emotional functioning of asthmatics. Moreover, incorporation of an individual style of coping with stress in the therapeutic process is necessary. PMID- 19308908 TI - [Analysis of solitary pulmonary nodules found in chest radiograms]. AB - INTRODUCTION: The detection of solitary pulmonary nodules (SPNs) has increased due to widespread use of computed tomography; nevertheless, chest radiographs still remain the basic routine examination. The aim of the study was to estimate the detection of SPNs in routine chest X-rays in hospitalized patients and to assess the incidence of malignancy in newly diagnosed SPNs. MATERIAL AND METHODS: We analyzed 5,726 routine chest radiographs of patients admitted to the Department of Internal Diseases, Pneumology and Allergology in 2004 and 2005. Most of the patients were admitted to hospital due to emergency reasons. The malignant nature of the nodules was confirmed by pathological examination. The nature of benign nodules was confirmed either by pathological examination or based on radiological criteria: no growth within 2 years of radiological follow up, regression in control radiograms or CT scans, benign pattern of calcification. RESULTS: Among the 5,726 radiograms we found 116 newly diagnosed SPNs (2.2%). Twenty-four nodules (21%) were malignant: NSCLC in 21 cases and metastases in 3 cases. Fifty-one nodules (44%) were benign. In 19 patients (16%) SPNs proved to be artefacts or erroneously interpreted extrathoracic lesions. In 22 cases (19%) there was no final diagnosis (lack of data, diagnostic procedure renunciation). CONCLUSION: The incidence of newly detected SPNs in chest X-rays was 2.2%. Most SPNs were benign. About 21% of SPNs were diagnosed as malignant. PMID- 19308909 TI - [Omalizumab, recombinant humanized monoclonal antibody anti-IgE -- new fields of studies on the therapeutic indications]. AB - Omalizumab is the monoclonal antibody against IgE, which blocks the binding of IgE to receptors on effector cells and this way blocks or decreases allergic reaction and asthma symptoms. Nowadays omalizumab is registered in the USA and European Union including Poland for treatment of patients with severe persistent, uncontrolled asthma. It is very effective in the reduction of asthma exacerbations, hospitalizations and emergency visits, it decreases intensity of symptoms and improves the quality of life. But omalizumab has more extensive action; given subcutaneously it has systemic effect. The article is the review of the studies and case reports on usage of omalizumab in new indications. Some authors' own experiences in this field are also presented. PMID- 19308910 TI - [Angiogenesis in interstitial lung diseases]. AB - Interstitial lung diseases (ILD) constitute a large group of disorders characterized by various etiology and pathogenesis. Inflammation and pulmonary fibrosis are the most important processes in the course of ILD. Disease causes the decrease of the gas diffusion in the lungs and provokes hypoxia. Chronic inflammation and hypoxia are strong stimulus of neovascularization. Neoangiogenesis is a principal response of vessels to inflammation. The critical importance of tumor angiogenesis in the development and metastatic spread of tumors is proved. Relations of ILD with neoplasma have been observed. Neovascularization takes an important role in wound healing allowing the cells to flow into damaged structures. Recently, pulmonary fibrosis has been deemed to result from abnormal wound healing in the lung in response to injury to the alveolar epithelium. Angiogenesis participates in pathogenesis of idiopathic pulmonary fibrosis. More and more data suggest the role of angiogenesis in pathogenesis of other ILDs, such as granulomatosis, fibrosis and vasculitis. The mechanism of angiogenesis in ILD is not clear yet. New data concerning participation of neoangiogenesis in pathogenesis of ILD created target for new drugs. Thalidomide, a strong antiangiogenic drug was used successfully in the some cases of ILD. PMID- 19308911 TI - [Tuberculosis after renal transplantation]. AB - Renal transplant recipients carry a relatively high risk of developing tuberculosis (TB). In most cases, active TB is the result of reactivation of a latent infection and is located in the lungs. In these patients, clinical presentation of TB can often be atypical and there is a high risk of dissemination and high mortality rates. Therefore, the use of invasive procedures for proper diagnosis is recommended, as well as anti-tuberculosis therapy instituted whenever there is a strong suspicion of TB on clinical grounds, even without microbiological evidence. The treatment of active TB in renal transplant recipients should be the same as in the general population. To avoid graft rejection, blood levels of calcineurin inhibitors should be monitored closely. Prophylaxis is recommended for high-risk patients. PMID- 19308912 TI - [Molecular mechanisms of inflammation in chronic obstructive pulmonary disease]. PMID- 19308913 TI - Physical activity in patients with COPD: the role of pulmonary rehabilitation. PMID- 19308914 TI - [Early diagnosis of COPD -- rationale, methods and effects]. PMID- 19308915 TI - Novel concepts in the pharmacotherapy of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is highly prevalent and will continue to be an increasing cause of morbidity and mortality worldwide. COPD is now viewed under a new paradigm as preventable and treatable. In addition, it has become accepted that COPD is not solely a pulmonary disease but also one with important measurable systemic consequences. It follows, that patients diagnosed with COPD have to be comprehensively evaluated to determine the extent of disease so that therapy can be adequately individualized. We now know that smoking cessation, oxygen for hypoxemic patients, lung reduction surgery for selected patients with emphysema, and non-invasive ventilation during severe exacerbations have an impact on mortality. The completion of well planned pharmacological trials have shown the importance of patient centered outcomes and the possible impact on mortality and rate of decline of lung function. This monograph presents an update on the pharmacological therapy of COPD. The future for patients with COPD is bright as primary and secondary prevention of smoking becomes more effective and air quality improves. In addition, current research will unravel the pathogenesis, clinical and phenotypic manifestations of COPD thus providing exciting therapeutic targets. Ultimately, the advent of newer and more effective therapies will lead to a decline in the contribution of this disease to poor world health. PMID- 19308916 TI - [Common variable immunodeficiency in a patient with suspected sarcoidosis]. AB - Common variable immunodeficiency is a primary immunodeficiency disease, characterized by hypogammaglobulinemia, low serum immunoglobulin concentrations, and recurrent bacterial infections of the respiratory and gastrointestinal tracts. We report on a 33-year-old patient with suspected sarcoidosis, diagnosed on the basis of an open lung biopsy, who was admitted to the National Tuberculosis and Lung Diseases Research Institute because of severe pneumonia and streptococcal sepsis. During diagnostics based on typical, clinical and laboratory features, CVID was diagnosed. The antibiotic treatment was successfully administered and the patient was directed to supplementary treatment. PMID- 19308919 TI - A Bayesian approach to joint analysis of longitudinal measurements and competing risks failure time data. AB - In this paper, we develop a Bayesian method for joint analysis of longitudinal measurements and competing risks failure time data. The model allows one to analyze the longitudinal outcome with nonignorable missing data induced by multiple types of events, to analyze survival data with dependent censoring for the key event, and to draw inferences on multiple endpoints simultaneously. Compared with the likelihood approach, the Bayesian method has several advantages. It is computationally more tractable for high-dimensional random effects. It is also convenient to draw inference. Moreover, it provides a means to incorporate prior information that may help to improve estimation accuracy. An illustration is given using a clinical trial data of scleroderma lung disease. The performance of our method is evaluated by simulation studies. PMID- 19308920 TI - Membrane permeability and antimicrobial kinetics of cecropin P1 against Escherichia coli. AB - The interaction of cecropin P1 (CP1) with Escherichiacoli was investigated to gain insight into the time-dependent antimicrobial action. Biophysical characterizations of CP1 with whole bacterial cells were performed using both fluorescent and colorimetric assays to investigate the role of membrane permeability and lipopolysaccharide (LPS) binding in lytic behavior. The kinetics of CP1 growth inhibition assays indicated a minimal inhibitory concentration (MIC) of 3 microM. Bactericidal kinetics at the MIC indicated rapid killing of E.coli (<30 min). Membrane permeability studies illustrated permeation as a time dependent event. Maximum permeability at the MIC occurred within 30 min, which correlates to the bactericidal action. Further investigation showed that the immediate permeabilizing action of CP1 is concentration-dependent, which correlates to the concentration-dependent nature of the inhibition assays. At the MIC and above, the immediate permeability was significant enough that the cells could not recover and exhibit growth. Below the MIC, immediate permeability was evident, but the level was insufficient to inhibit growth. Dansyl polymyxin B displacement studies showed LPS binding is essentially the same at all concentrations investigated. However, it does appear that only the immediate interaction is important, because binding continued to increase over time beyond cell viability. Our studies correlated CP1 bactericidal kinetics to membrane permeability suggesting CP1 concentration-dependent killing is driven by the extent of the immediate permeabilizing action of the peptide. PMID- 19308921 TI - A Ru-Hbpp-based water-oxidation catalyst anchored on rutile TiO2. AB - Two organic ligands based on bis-(2-pyridyl)pyrazole (Hbpp) functionalized with a para-methylenebenzoic acid (Hbpp-R(a)) or its ester derivative (Hbpp-R(e)) were prepared and characterized. The ester-functionalized ligand was then used to prepare a series of related dinuclear ruthenium complexes of general formula [Ru(II)2(L-L)(bpp-R(n))(trpy)2](m+) (L-L=mu-Cl, mu-acetato, or (H2O)2; n=e or a; trpy=2,2':6',2''-terpyridine; m=2 or 3). The complexes were characterized in solution by 1D and 2D NMR spectroscopy, UV/Vis spectroscopy, and electrochemical techniques. The [Ru(II)2(mu-Cl)(bpp-R(e))(trpy)2](PF6)(2) complex was further characterized in the solid state by X-ray diffraction. The complexes containing the free carboxylic acid ligand were anchored onto rutile TiO2 and treated with 0.1 M triflic acid solution to generate the homologous water-oxidation catalysts TiO(2)-[Ru(II) (2)(H2O)(2)(bpp-R(a))(trpy)2]2+. This new hybrid material catalytically oxidizes water to molecular oxygen in a heterogeneous manner using Ce(IV) as chemical oxidant. The generation of molecular oxygen is accompanied by the formation of carbon dioxide as well as some leaching of the Ru catalyst. PMID- 19308922 TI - Tacrine-melatonin hybrids as multifunctional agents for Alzheimer's disease, with cholinergic, antioxidant, and neuroprotective properties. AB - Tacrine-melatonin hybrids were designed and synthesized as new multifunctional drug candidates for Alzheimer's disease. These compounds may simultaneously palliate intellectual deficits and protect the brain against both beta-amyloid (A beta) peptide and oxidative stress. They show improved cholinergic and antioxidant properties, and are more potent and selective inhibitors of human acetylcholinesterase (hAChE) than tacrine. They also capture free radicals better than melatonin. Molecular modeling studies show that these hybrids target both the catalytic active site (CAS) and the peripheral anionic site (PAS) of AChE. At sub-micromolar concentrations they efficiently displace the binding of propidium iodide from the PAS and could thus inhibit A beta peptide aggregation promoted by AChE. Moreover, they also inhibit A beta self-aggregation and display neuroprotective properties in a human neuroblastoma line against cell death induced by various toxic insults, such as A beta(25-35), H(2)O(2), and rotenone. Finally, they exhibit low toxicity and may be able to penetrate the central nervous system according to an in vitro parallel artificial membrane permeability assay for the blood-brain barrier (PAMPA-BBB). PMID- 19308923 TI - Bacterial alarmone, guanosine 5'-diphosphate 3'-diphosphate (ppGpp), predominantly binds the beta' subunit of plastid-encoded plastid RNA polymerase in chloroplasts. AB - It's alarming: Bacterial alarmone guanosine 5'-diphosphate 3'-diphosphate (ppGpp), which is a key regulatory molecule that controls the stringent response, also exists in chloroplasts of plant cells. Cross-linking experiments with 6 thioguanosine 5'-diphosphate 3'-diphosphate (6-thioppGpp) and chloroplast RNA polymerase indicate that ppGpp binds the beta' subunit of plastid-encoded plastid RNA polymerase that corresponds to the Escherichia coli beta' subunit. Chloroplasts, which are thought to have originated from cyanobacteria, have their own genetic system that is similar to that of the bacteria from which they were derived. Recently, bacterial alarmone guanosine 5'-diphosphate 3'-diphosphate (ppGpp, 1), a key regulatory molecule that controls the stringent response, was identified in the chloroplasts of plant cells. Similar to its function in bacteria, ppGpp inhibits chloroplast RNA polymerase; this suggests that ppGpp mediates gene expression through the stringent response in chloroplasts. However, a detailed mechanism of ppGpp action in chloroplasts remains elusive. We synthesized 6-thioguanosine 5'-diphosphate 3'-diphosphate (6-thioppGpp) as a photoaffinity probe of ppGpp; this probe thus enabled the investigation of ppGpp binding to chloroplast RNA polymerase. We found that 6-thioppGpp, as well as ppGpp, inhibits chloroplast RNA synthesis in vitro in a dose-dependent manner. Cross-linking experiments with 6-thioppGpp and chloroplast RNA polymerase indicated that ppGpp binds the beta' subunit (corresponding to the Escherichia coli beta' subunit) of plastid-encoded plastid RNA polymerase composed of alpha, beta, beta', beta'', and sigma subunits. Furthermore, ppGpp did not inhibit transcription in plastid nucleoids prepared from tobacco BY-2 cells; this suggests that ppGpp does not inhibit nuclear-encoded plastid RNA polymerase. PMID- 19308924 TI - Rational biosynthetic engineering for optimization of geldanamycin analogues. AB - Tailor made: We report the rational biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. Rational biosynthetic engineering allowed the generation of geldanamycin derivatives, such as DHQ3 illustrated in the figure, which had superior pharmacological properties in comparison to the parent compound. A rational biosynthetic engineering approach was applied to the optimization of the pharmacological properties of the benzoquinone ansamycin, geldanamycin. Geldanamycin and its natural or semisynthetic derivatives have the potential to serve as anticancer chemotherapeutic agents. However, these first-generation Hsp90 inhibitors share an unfavorable structural feature that causes both reduced efficacy and toxicity during clinical evaluation. We report the rationally designed biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. A 15-hydroxyl-17-demethoxy non quinone analogue, DHQ3, exhibited stronger inhibition of Hsp90 ATPase activity (4.6-fold) than geldanamycin. Taken together, the results of the present study indicate that rational biosynthetic engineering allows the generation of derivatives of geldanamycin with superior pharmacological properties. PMID- 19308925 TI - Probing mutation-induced structural perturbations by refinement against residual dipolar couplings: application to the U4 spliceosomal RNP complex. AB - Confident interpretation of biochemical experiments performed with mutated proteins relies on verification of the integrity of the mutant structures. We present a simple and rapid refinement protocol for comparing the structures of mutated and wild-type proteins. Our approach involves measurement of residual dipolar couplings, and only requires assignment of the backbone resonances of the mutant species. We demonstrate application of the protocol to a mutant of the 15.5K protein, a core component of the U4 spliceosomal ribonucleoprotein (RNP) complex. Confirmation of the unperturbed structure of the mutated protein prompted re-examination of a previous mutagenesis study and indicated that the interpretation of mutant binding affinities in terms of direct interfacial contacts should be applied with caution. PMID- 19308926 TI - Biomolecular oxidative damage activated by enzymatic logic systems: biologically inspired approach. AB - Systems that perform oxidative damage to biomolecules through catalytic cascades in the presence of iron-redox labile species were activated by enzymatic logic gates that process chemical input signals according to built-in logic operations. AND/OR enzymatic logic gates were composed of glucose oxidase (GOx) and GOx/esterase, respectively. The AND/OR logic functions of the enzyme gates were activated by application of glucose-oxygen and glucose-ethyl acetate input signals, respectively. The enzymatic logic gates, upon activation by specific patterns of the chemical input signals, produced acidic solutions and triggered release of redox labile iron species from a complex that is unstable under acidic conditions. This resulted in the activation of a catalytic cascade, which produced reactive oxygen species (ROS) and subsequently yielded oxidative damage in biomolecules. Functional integration of the enzyme-based logic systems with the catalytic redox cascade that performs damage in biomolecules on demand is a first step towards "smart" systems capable of programmed detection, identification, and neutralization of potential biohazards. PMID- 19308927 TI - In vivo modification of native carrier protein domains. AB - Carrier proteins are central to the biosynthesis of primary and secondary metabolites in all organisms. Here we describe metabolic labeling and manipulation of native acyl carrier proteins in both type I and II fatty acid synthases. By utilizing natural promiscuity in the CoA biosynthetic pathway in combination with synthetic pantetheine analogues, we demonstrate metabolic labeling of endogenous carrier proteins with reporter tags in Gram-positive and Gram-negative bacteria and in a human carcinoma cell line. The highly specific nature of the post-translational modification that was utilized for tagging allows for simple visualization of labeled carrier proteins, either by direct fluorescence imaging or after chemical conjugation to a fluorescent reporter. In addition, we demonstrate the utility of this approach for the isolation and enrichment of carrier proteins by affinity purification. Finally, we use these techniques to identify a carrier protein from an unsequenced organism, a finding that validates this proteomic approach to natural product biosynthetic enzyme discovery. PMID- 19308928 TI - Photocrosslinking of RNA and photoMet-containing amphiphilic alpha-helical peptides. AB - Discovering RNA-protein interactions: A library of photoMet-containing peptides was synthesized by using an Arg- and Leu-rich alpha-helical amphiphilic peptide. Irradiation of mixtures of these peptides and Rev-responsive element (RRE) hairpin RNA promoted formation of covalent adducts. Analysis of one adduct showed that U26 in the bulged stem is responsible for covalent bond formation with the carbene intermediate. This strategy can provide important structural information about RNA-peptide interactions. PMID- 19308929 TI - Sonographic imaging of urinoma. PMID- 19308930 TI - Amniotic fluid 'sludge' detected in patients with subchorionic hematoma: a report of two cases. AB - Amniotic fluid 'sludge' is defined as the presence of dense aggregates of particulate matter in close proximity to the internal cervical os. It is of clinical significance in asymptomatic patients at high risk for spontaneous delivery, and in patients with preterm labor and intact membranes. Subchorionic hematoma is another ultrasound finding that is associated with a higher incidence of threatened miscarriage and preterm delivery. We report two cases of occurrence of amniotic fluid sludge in patients with previously detected large subchorionic hematoma. In the first case subchorionic hematoma and amniotic fluid sludge were detected by ultrasonography at 13 + 1 and 18 + 6 weeks' gestation, respectively, followed by preterm premature rupture of membranes, placental abruption and emergency Cesarean section. In the second case subchorionic hematoma and amniotic fluid sludge were detected by ultrasound at 11 + 3 and 15 + 5 weeks' gestation, respectively, followed by miscarriage with histological chorioamnionitis. The coincidence of subchorionic hematoma and amniotic fluid sludge in these cases points to a possible connection between these two significant ultrasound findings. PMID- 19308931 TI - Retinal coloboma: prenatal diagnosis using a new technique, the 'virtual fetal eyeground'. PMID- 19308932 TI - Immobilization strategies for small molecule, peptide and protein microarrays. AB - Protein, peptide and small molecule microarrays are valuable tools in biological research. In the last decade, substantial progress has been achieved to make these powerful technologies more reliable and available for researchers. This review describes chemical preparation methods for these microarrays with focus on site-selective and bioorthogonal immobilization reactions, particularly the Staudinger ligation and the thiol-ene reaction. In addition, the application of peptide microarrays, which were prepared by Staudinger ligation, to substrate specificity mapping is illustrated. PMID- 19308933 TI - A multivariate CAR model for improving the estimation of relative risks. AB - Disease mapping studies have been widely performed at univariate level, that is considering only one disease in the estimated models. Nonetheless, simultaneous modelling of different diseases can be a valuable tool both from the epidemiological and from the statistical point of view. In this paper we propose a model for multivariate disease mapping that generalizes the univariate conditional auto-regressive distribution. The proposed model is proven to be an effective alternative to existing multivariate models, mainly because it overcome some restrictive hypotheses underlying models previously proposed in this context. Model performances are checked via a simulation study and via application to a case study. PMID- 19308934 TI - Prestimulus alpha and mu activity predicts failure to inhibit motor responses. AB - Do certain brain states predispose humans to commit errors in monotonous tasks? We used MEG to investigate how oscillatory brain activity indexes the brain state in subjects performing a Go-noGo task. Elevated occipital alpha and sensorimotor mu activity just prior to the presentation of the stimuli predicted an upcoming error. An error resulted in increased frontal theta activity and decreased posterior alpha activity. This theta increase and alpha decrease correlated on a trial-by-trial basis reflecting post-error functional connectivity between the frontal and occipital regions. By examining the state of the brain before a stimulus, we were able to show that it is possible to predict lapses of attention before they actually occur. This supports the case that the state of the brain is important for how incoming stimuli are processed and for how subjects respond. PMID- 19308935 TI - Location, location, (ChIP-)location! Mapping chromatin landscapes one immunoprecipitation at a time. AB - A small fraction of the typical animal genome (<5% in humans) codes for the organism's collection of proteins, yet the study of protein coding sequences dominated the early years of genomics research. In the decade since the sequencing of complete eukaryotic genomes, however, genomic techniques have shed a great deal of light on the non-coding DNA making up the remainder. A single molecular technique, Chromatin Immuno-Precipitation (ChIP) location analysis, has had a profound impact and has made possible the study of an incredible range of biology. This issue of The Journal of Cellular Biochemistry aims to put into context advancements made possible by the ChIP-location revolution, while at the same time highlighting some of the most important technical aspects and challenges along with some of the work yet to come. PMID- 19308936 TI - Functional p53 in cells contributes to the anticancer effect of the cyclin dependent kinase inhibitor roscovitine. AB - Inhibitors of cyclin-dependent kinases (CDKs) undergoing clinical trials as anticancer agents usually target several CDKs in cells. Some of them are also able to increase cellular levels of p53 protein and to activate p53-regulated transcription. To define the role of p53 in the anticancer effect of selective CDK inhibitors, two related compounds roscovitine and olomoucine II were studied. Roscovitine differs functionally from its congener olomoucine II only in the selectivity towards transcriptional CDK9. Action of both compounds on proliferation, cell-cycle progression, and apoptosis was examined in RPMI-8226 cells expressing the temperature-sensitive mutant of p53 and in MCF-7 cells with wild-type p53. Both compounds blocked proliferation, decreased phosphorylation of RNA polymerase II, downregulated antiapoptotic protein Mcl-1 in both cell lines in a dose-dependent manner, and also activated p53 in MCF-7 cells. Moreover, we showed that the anticancer efficiency of CDK inhibitors was enhanced by active p53 in RPMI-8226 cells kept at permissive temperature, where downregulation of Mcl-1, fragmentation of PARP-1, and increased caspase-3 activity was detected with lower doses of the compounds. The results confirm that functional p53 protein may enhance the anticancer activity of roscovitine that could be beneficial for anticancer therapy. PMID- 19308937 TI - Multicomponent reactions for the synthesis of complex piperidine scaffolds. PMID- 19308938 TI - Correction of errors during protein synthesis. AB - Quality control: The incorporation of a wrong amino acid into a growing polypeptide chain induces a correction step in which the release factor (RF1) hydrolyzes the peptide from the incorrectly matched peptidyl-tRNA (see picture). The nascent erroneous polypeptide is released from the ribosome and degraded. PMID- 19308939 TI - Palladium-catalyzed oxidative intermolecular difunctionalization of terminal alkenes with organostannanes and molecular oxygen. AB - A cationic palladium complex catalyzes the title transformations, which are thought to proceed via a pi-allyl or pi-benzyl intermediate. The regioselectivity of the reaction (1,2- or 1,1-difunctionalization) depends on the type of terminal double bond (conjugated or nonconjugated) in the substrate (see scheme) and appears to be controlled by the relative rates of beta-hydride elimination and transmetalation. DMA=dimethylacetamide, Tf=triflyl. PMID- 19308940 TI - Solution structure of a locked nucleic acid modified quadruplex: introducing the V4 folding topology. AB - Sharp curves: The structure of a locked nucleic acid modified telomeric sequence from Oxytricha nova displays a remarkable folding topology, distinct from the native O. nova quadruplex. Each guanine stretch folds back in a V-shaped turn that puts the first and fourth guanines in the same tetrad, looping over a tetrad with a sharp turn in the DNA backbone, showing how subtle interplay between sequence and conformation defines the folding topology. PMID- 19308941 TI - Selective adsorption of DNA on chiral surfaces: supercoiled or relaxed conformation. AB - The right fit: Plasmid DNA molecules show chirality-dependent interaction with gold surfaces modified by L and D N-isobutyrylcysteine. Relaxed DNA molecules have a stronger interaction and adsorption on the L surface, while their counterparts on the D surface maintain a supercoiled conformation, indicating a weak interaction (see picture). PMID- 19308942 TI - Hydrogen bonding in protic ionic liquids: reminiscent of water. AB - Similarities and differences: Far-infrared spectra of protic ionic liquids could be assigned to intermolecular bending and stretching modes of hydrogen bonds. The characteristics of the low-frequency spectra resemble those of water. Both liquids form three-dimensional network structures, but only water is capable of building tetrahedral configurations. EAN: ethylammonium nitrate, PAN: propylammonium nitrate, DMAN: dimethylammonium nitrate. PMID- 19308944 TI - Perfectionism and eating attitudes in Portuguese students: a longitudinal study. AB - AIM: To investigate the role of perfectionism in the development of disordered eating behaviours. METHOD: 382 female university students completed the Hewitt & Flett MPS and the EAT-40 at baseline, and 1 year after (T1) and 206 2 years later (T2). RESULTS: Perfectionism at baseline was significantly associated with long term abnormal eating attitudes/behaviours. Self-Oriented Perfectionism (SOP) and Socially Prescribed Perfectionism (SPP) were significant predictors of disordered eating behaviours. Regression analysis revealed that SOP at baseline was predictive of Diet Concerns and overall eating disturbance (EAT total score), at T1 and T2. SPP was a significant predictor of Social Pressure to Eat at T1 and T2 and of Bulimic Behaviours only at T1. CONCLUSION: Our findings contribute to a more clear understanding of the association between perfectionism and eating disorders. SOP and SPP were prospectively associated with abnormal eating attitudes/behaviours and SOP was found to be predictive of diet concerns and overall eating disturbance. PMID- 19308943 TI - Activation of GABA-A receptor ameliorates homocysteine-induced MMP-9 activation by ERK pathway. AB - Hyperhomocysteinemia (HHcy) is a risk factor for neuroinflammatory and neurodegenerative diseases. Homocysteine (Hcy) induces redox stress, in part, by activating matrix metalloproteinase-9 (MMP-9), which degrades the matrix and leads to blood-brain barrier dysfunction. Hcy competitively binds to gamma aminbutyric acid (GABA) receptors, which are excitatory neurotransmitter receptors. However, the role of GABA-A receptor in Hcy-induced cerebrovascular remodeling is not clear. We hypothesized that Hcy causes cerebrovascular remodeling by increasing redox stress and MMP-9 activity via the extracellular signal-regulated kinase (ERK) signaling pathway and by inhibition of GABA-A receptors, thus behaving as an inhibitory neurotransmitter. Hcy-induced reactive oxygen species production was detected using the fluorescent probe, 2'-7' dichlorodihydrofluorescein diacetate. Hcy increased nicotinamide adenine dinucleotide phosphate-oxidase-4 concomitantly suppressing thioredoxin. Hcy caused activation of MMP-9, measured by gelatin zymography. The GABA-A receptor agonist, muscimol ameliorated the Hcy-mediated MMP-9 activation. In parallel, Hcy caused phosphorylation of ERK and selectively decreased levels of tissue inhibitors of metalloproteinase-4 (TIMP-4). Treatment of the endothelial cell with muscimol restored the levels of TIMP-4 to the levels in control group. Hcy induced expression of iNOS and decreased eNOS expression, which lead to a decreased NO bioavailability. Furthermore muscimol attenuated Hcy-induced MMP-9 via ERK signaling pathway. These results suggest that Hcy competes with GABA-A receptors, inducing the oxidative stress transduction pathway and leading to ERK activation. PMID- 19308945 TI - The prevalence, incidence and development of eating disorders in Finnish adolescents: a two-step 3-year follow-up study. AB - OBJECTIVE: To investigate the prevalence, incidence and development of eating disorders and subclinical eating pathology. METHOD: A two-step three-year follow up study on eating disorders in adolescence (N = 595) was conducted in western Finland. A screening questionnaire followed by a semi-structured interview was used to determine the prevalence, incidence and development of eating disorders. RESULTS: The lifetime prevalence rates for females age 18 were 2.6% for anorexia nervosa (AN), 0.4 for bulimia nervosa (BN), 7.7% for AN-NOS, 1.3% for BN-NOS and 8.5% for subclinical eating disorders. No prevalent case of DSM-IV eating disorders was found among the male participants. The incidence rate of any eating disorder in females age 15-18 was 1641 per 100 000 person-years. CONCLUSION: Eating disorders are relatively common in female adolescents. As many as one in five adolescent females are or have been struggling with eating disorder related issues during their teenage years. PMID- 19308946 TI - Factors related to motivation for change in adolescents with eating disorders. AB - OBJECTIVE: To examine the association between motivation for change and eating disorder symptom severity, the quality of the parent-adolescent relationship and depressive symptom severity in adolescents with eating disorders. METHOD: Fifty four female adolescents with eating disorders (mean age = 15.8 years, SD = 1.6) participated in this research prior to beginning outpatient treatment. Participants completed the Motivational Stages of Change for Adolescents Recovering from an Eating Disorder, the Eating Disorders Inventory-2, the Relations with Parents scale from the Behavioral Assessment System for Children and Adolescents Self-report and the Child Depression Inventory. RESULTS: Greater motivation for change was associated with less body dissatisfaction, more adaptive parent-adolescent relationships and fewer depressive symptoms. The association between motivation for change and adolescents' perception of their relationship with their parents remained significant even after controlling for age, body dissatisfaction and depressive symptoms. CONCLUSION: Involvement of families in treatment for adolescents with eating disorders may serve to enhance their motivation to recover. PMID- 19308947 TI - Metabonomic identification of novel biomarkers in doxorubicin cardiotoxicity and protective effect of the natural antioxidant oleuropein. AB - Doxorubicin (DXR) is a commonly used antineoplastic agent; however, its use is limited due to cardiotoxicity. Oxidative stress and consequent alterations of cardiac energetics are involved in the development of DXR toxicity. Oleuropein (Oleu) is a phenolic antioxidant, present in olive tree, reported to confer protection against DXR cardiotoxicity. In this study, NMR based-metabonomics was applied to characterize the metabolic profile of the acute DXR cardiotoxicity in rats and to evaluate the metabolic alterations conferred by co-treatment with Oleu. Wistar rats were divided into six groups and treated as follows: control group with a single injection of 2 mL normal saline intraperitoneally (i.p.), DXR group with a single dose of 20 mg/kg, i.p and DXR plus Oleu groups with 20mg/kg DXR i.p., and 100 or 200 mg/kg/BW of Oleu i.p. for 5 or 3 consecutive days starting either 2 days before or on the day of DXR administration. Hearts were excised 72 h after DXR treatment and (1)H-NMR spectra of aqueous myocardium extracts were recorded. Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis (PLS-DA) revealed differences in the metabolic profile between control and DXR attributed to several metabolites. A number of them were quantified by integration of the NMR spectra. Myocardial levels of acetate and succinate were increased in DXR compared to controls, while branched amino acids were decreased. These results correlate with nonenzymatic conversion of pyruvate to acetate and of alpha-ketoglutarate to succinate by DXR free radicals. Oleu completely restored the changes of metabolites to the normal levels. Acetate and succinate constitute novel biomarkers related to DXR, and Oleu treatment aids the compensation of distressed energy metabolic pathways. PMID- 19308948 TI - An anterior-lateral thigh perforator flap on a recipient brachial-radial vein graft for complex wound reconstruction: a case report. AB - A case of challenging microsurgical reconstruction of a difficult defect in a radiated upper limb is reported. A difficult wound, with tendon and bone exposition, developed on the dorsum of the forearm in a 76-year-old patient; she had been radiated since almost 50 years and her left hand had also been revascularized twice with venous grafts between the humeral artery and the superficial palmar arch. After failure of a local flap, an anterior-lateral thigh perforator flap was successfully transferred with end-to-side anastomoses on the arterialized venous graft. Up to date follow-up shows a good outcome. The Authors discuss the case and review the indications for microsurgical reconstruction in difficult wounds after radiation and ischemic limb conditions. PMID- 19308949 TI - Three-dimensional reconstruction and visualization of the median nerve from serial tissue sections. AB - The purpose of this study was to definitively implement the three-dimensional visualization of sensory and motor fascicles in the human median nerve by means of acetylcholinesterase (AChe) histochemical staining and under the assistance of the computer technology. One fresh human median nerve was harvested from a male adult cadaver. The median nerve was fixed at a special holder. Then, the whole holder was embedded and rapidly frozen in the liquid nitrogen. The processed median nerve was then cut coronally every 100 microm at a 20 microm thickness along its long axis in a sliding freezing microtome. The total number of sections was 4,650 slices. All sections were stained with the AChe histochemical method. The stained sections were scanned and saved as Joint Photographic Experts Group files. These images with positively and negatively stained sections were acquired to an Intel dual Pentium computer. The Adobe Photoshop CS2 software was used to compare the reference points of images before and after staining. The two dimensional intraneural microstructure database of median nerve was then acquired. A software of 3D nerve visualization system was developed. With the 3D nerve visualization system, the 3D visualization result of intraneural microstructure of median nerve was created. The findings may provide more accurate and detailed anatomic information for nerve repairs, specifically for the fascicular nerve repairs. The 3D nerve visualization technique may have potential for future studies of topography of peripheral nerve. (c) 2009 Wiley Liss, Inc. Microsurgery, 2009. PMID- 19308950 TI - Effect of low-intensity pulsed ultrasound on the expression of neurotrophin-3 and brain-derived neurotrophic factor in cultured Schwann cells. AB - It is generally known that low-intensity pulsed ultrasound (LIPUS) accelerates peripheral nerve tissue regeneration. However, the precise cellular mechanism involved is still unclear. The purpose of this study was to determine how the Schwann cells respond directly to LIPUS stimuli. Thus, we investigated the effect of LIPUS on cell proliferation, neurotrophin-3 (NT-3), and brain-derived neurotrophic factor (BDNF) mRNA expression in rat Schwann cells. Schwann cells were enzymatically isolated from postnatal 1-3 day rat sciatic nerve tissue and cultured in the six-well plate. The ultrasound was applied at a frequency of 1 MHz and an intensity of 100 mW/cm(2) spatial average temporal average for 5 minutes/day. The control group was cultured in the same way but without the administration of ultrasound. Immunohistochemistry demonstrated that more than 98% of the experimental and control cells were positive for S-100, NT-3, and BDNF. With 5-bromo-2'-deoxyuridine (BrdU) assay, the stimulated cells also exhibited an increase in the rate of cell proliferation on days 4, 7, 10, and 14. Further investigation found that mRNA expression of NT-3 was significantly upregulated in experimental groups compared with the control 14 days after the LIPUS stimulation (the ratio of NT-3/beta-actin was 0.56 +/- 0.13 vs. 0.41 +/- 0.09, P < 0.01), whereas the mRNA expression of BDNF was significantly downregulated in experimental groups compared with the control (the ratio of BDNF/beta-actin was 0.51 +/- 0.05 vs. 0.60 +/- 0.08, P < 0.05). These results demonstrated that the application of LIPUS promotes cell proliferation and NT-3 gene expression in Schwann cells, and involved in the alteration of BDNF gene expression. PMID- 19308951 TI - Mass spectrometric study of peptides secreted by the skin glands of the brown frog Rana arvalis from the Moscow region. AB - A high-performance liquid chromatography nano-electrospray ionization Fourier transform mass spectrometry (HPLC/nanoESI-FTMS) approach involving recording of collision-activated dissociation (CAD) and electron-capture dissociation (ECD) spectra of an intact sample and two its modifications after performic oxidation and reduction followed by carboxamidomethylation helps to establish peptide profiles in the crude secretion of frog species at mid-throughput level, including de novo sequencing. The proposed derivatization procedures allow increasing of the general sequence coverage in the backbone, providing complementary information and, what is more important, reveal the amino acid sequence in the cystine ring ('rana box'). Thus purely mass spectrometric efficient sequencing becomes possible for longer than usual proteolytic peptides. Seventeen peptides belonging to four known families were identified in the secretion of the European brown frog Rana arvalis inhabiting the Moscow region in Russia. Ranatuerins, considered previously a unique feature of the North American species, as well as a new melittin-related peptide, are worth special mention. The developed approach was previously successfully used for the identification of peptides in the skin secretion of the Caucasian green frog Rana ridibunda. PMID- 19308952 TI - Porosity of the wall of a Neurolac nerve conduit hampers nerve regeneration. AB - One way to improve nerve regeneration and bridge longer nerve gaps may be the use of semipermeable/porous conduits. With porosity less biomaterial is used for the nerve conduit. We evaluated the short-term effects of porous Neurolac nerve conduits for in vivo peripheral nerve regeneration. In 10 male Black Hooded rats, a gap of 10 mm was bridged by a porous Neurolac nerve conduit. Evaluation point ranged from 3 to 12 weeks. The sciatic nerve function was not measurable due to automutilation and flexion contractures. The gait-stance duration showed no improvement with time, indicating a disturbed walking pattern. The nerve guides showed very fast degradation with swelling, fragmentation, and collapse. Furthermore, a severe foreign body reaction occurred. Nerve regeneration was severely hampered. This study showed no beneficial effects of porous Neurolac nerve conduits when compared with previous findings with nonporous copolymeric nerve guides of a slightly different composition. PMID- 19308953 TI - Face resurfacing using a cervicothoracic skin flap prefabricated by lateral thigh fascial flap and tissue expander. AB - BACKGROUND: Resurfacing of facial massive soft tissue defect is a formidable challenge because of the unique character of the region and the limitation of well-matched donor site. In this report, we introduce a technique for using the prefabricated cervicothoracic skin flap for facial resurfacing, in an attempt to meet the principle of flap selection in face reconstructive surgery for matching the color and texture, large dimension, and thinner thickness (MLT) of the recipient. MATERIALS: Eleven patients with massive facial scars underwent resurfacing procedures with prefabricated cervicothoracic flaps. The vasculature of the lateral thigh fascial flap, including the descending branch of the lateral femoral circumflex vessels and the surrounding muscle fascia, was used as the vascular carrier, and the pedicles of the fascial flap were anastomosed to either the superior thyroid or facial vessels in flap prefabrication. A tissue expander was placed beneath the fascial flap to enlarge the size and reduce the thickness of the flap. RESULTS: The average size of the harvested fascia flap was 6.5 x 11.7 cm. After a mean interval of 21.5 weeks, the expanders were filled to a mean volume of 1,685 ml. The sizes of the prefabricated skin flaps ranged from 12 x 15 cm to 15 x 32 cm. The prefabricated skin flaps were then transferred to the recipient site as pedicled flaps for facial resurfacing. All facial soft tissue defects were successfully covered by the flaps. The donor sites were primarily closed and healed without complications. Although varied degrees of venous congestion were developed after flap transfers, the marginal necrosis only occurred in two cases. The results in follow-up showed most resurfaced faces restored natural contour and regained emotional expression. CONCLUSION: MLT is the principle for flap selection in resurfacing of the massive facial soft tissue defect. Our experience in this series of patients demonstrated that the prefabricated cervicothoracic skin flap could be a reliable alternative tool for resurfacing of massive facial soft tissue defects. (c) 2009 Wiley-Liss, Inc. Microsurgery, 2009. PMID- 19308954 TI - Effects of acetylo-L-carnitine in end-to-side neurorrhaphy: a pilot study. AB - PURPOSE: End-to-side (ETS) nerve repair allows for target-muscle reinnervation, with simultaneous preservation of donor-nerve function. Acetyl-L-carnitine (ALCAR) was shown to enhance axonal sprouting in early regeneration following transection and repair of the sciatic nerve in rodents. The purpose of this article was to determine the ability of ALCAR to enhance axonal regeneration in an ETS rodent model. METHOD: The right musculocutaneous nerve in 16 adult male Sprague-Dawley rats was transected to induce biceps muscle paralysis. The distal stump was then coapted by ETS neurorrhaphy through a perineurial window to the ipsilateral median nerve. Experimental groups received ALCAR for 1, 2, 3, and 4 weeks whereas controls received placebo. RESULTS: Weekly postoperative behavioral evaluations revealed increased functional return over control but the difference was not significant. Potentials from biceps were recorded from the third postoperative week in the experimental group and from the fourth week in the control group. Histomorphometric evaluations revealed higher musculocutaneous nerve axon counts, higher myelin thickness in the fourth postoperative week, and differences in the appearance and the number of motor-end-plates in the biceps in experimental versus control group. CONCLUSION: Intraperitoneal administration of ALCAR can expedite biceps muscle recovery in an ETS model by increasing the rate of axonal regeneration. Despite the morphological changes, no behavioral changes were noted and further studies are needed to confirm clinical efficacy of ALCAR for potential use in the development of therapeutic protocols. PMID- 19308955 TI - Reverse flow facial artery as recipient vessel for perforator flaps. AB - In perforator flaps, anastomosis between flap and recipient vessels in the neck area is often difficult due to small vessel diameter and short pedicle. The aim of this study was to investigate whether the retrograde flow of the distal, paramandibular part of the facial artery would provide sufficient pressure and size to perfuse perforator flaps. Before and after occlusion of the contralateral facial artery, retrograde and anterograde arterial pressure was measured on both sides of the facial artery in 50 patients. The values were compared with the mean systemic arterial pressure. Diameters of facial arteries in the paramandibular region and perforator flap vessels were evaluated by morphometry. Arterial pressure in the distal facial artery with retrograde flow was 76% of the systemic arterial pressure. The latter equaled approximately the anterograde arterial pressure in the proximal end of the facial artery. Mean arterial pressure of the facial arteries decreased after proximal occlusion of the contralateral facial artery, which was not significant (P = 0.09). Mean diameter of the distal facial arteries in the mandibular region was 1.6 mm (range 1.3-2.2 mm; standard deviation 0.3 mm; n = 50), that of the perforator flap arteries 1.3 mm (0.9-2.6 mm; 0.4 mm; n = 20). Facial arteries, based on reverse flow, successfully supported all 20 perforator flaps. Retrograde pulsatile flow in the distal facial artery sustains perforator flaps even if the contralateral facial artery is occluded. Proximity of the distal facial arteries to the defect compensates for short pedicles. Matching diameters of the arteries are ideal for end-to-end anastomosis. PMID- 19308956 TI - Repair of electrically injured median nerve with the aid of somatosensory evoked potential. AB - In repair of the high voltage electrical injuries of peripheral nerve, selection of the proper plane for nerve coaptation after debridement is important since the electrical current is inclined to pass through the nerve trunks due to the lower electric resistance. In this report, we present our experience on use of the somatosensory evoked potentials (SSEP) test to assist determination of range of the injured nerve debridement in the electrical injured nerve. From January 1997 to December 2007, 30 patients with the high voltage electrical injury in the wrist, having defect of median nerve, were treated with nerve grafting. In 18 patients, the nerve resection plane was selected with the assistance of SSEP. In 12 patients, the nerve resection plane was determined by the traditional gross examination. The average lengths of nerve defect after debridement were 10.4 cm in the procedure with SSEP assistance and 4.01 cm in the traditional procedure. The finger motion was reconstructed by the tendon graft connecting the proximal forearm muscle and the distal tendon of flexor disitorum profundus. In following up, the two-point discrimination of finger was examined. The grade of finger sensory recovery was found improved in the patients who underwent the nerve repair with SSEP assistance than that in the patients underwent treatment with the traditional method. The results suggested that aggressive debridement is recommended for the high with voltage electrical nerve injury. SSEP could be a valuable tool for determination of the range of nerve debridement. PMID- 19308957 TI - Temporal patterns of lipoperoxidation and antioxidant enzymes are modified in the hippocampus of vitamin A-deficient rats. AB - Animals can adapt their behavior to predictable temporal fluctuations in the environment through both, memory-and-learning processes and an endogenous time keeping mechanism. Hippocampus plays a key role in memory and learning and is especially susceptible to oxidative stress. In compensation, antioxidant enzymes activity, such as Catalase (CAT) and Glutathione peroxidase (GPx), has been detected in this brain region. Daily rhythms of antioxidant enzymes activity, as well as of glutathione and lipid peroxides levels, have been described in brain. Here, we investigate day/night variations in lipoperoxidation, CAT, and GPx expression and activity, as well as the temporal fluctuations of two key components of the endogenous clock, BMAL1 and PER1, in the rat hippocampus and evaluate to which extent vitamin A deficiency may affect their amplitude or phase. Holtzman male rats from control, vitamin A-deficient, and vitamin A-refed groups were sacrificed throughout a 24-h period. Daily levels of clock proteins, lipoperoxidation, CAT and GPx mRNA, protein, and activity, were determined in the rat hippocampus obtained every 4 or 5 h. Gene expression of RARalpha and RXRbeta was also quantified in the hippocampus of the three groups of rats. Our results show significant daily variations of BMAL1 and PER1 protein expression. Rhythmic lipoperoxidation, CAT, and GPx, expression and activity, were also observed in the rat hippocampus. Vitamin A deficiency reduced RXRbeta mRNA level, as well as the amplitude of BMAL1 and PER1 daily oscillation, phase-shifted the daily peak of lipoperoxidation, and had a differential effect on the oscillating CAT and GPx mRNA, protein, and activity. Learning how vitamin A deficiency affects the circadian gene expression in the hippocampus may have an impact on the neurobiology, nutritional and chronobiology fields, emphasizing for the first time the importance of nutritional factors, such as dietary micronutrients, in the regulation of circadian parameters in this brain memory-and-learning-related region. PMID- 19308958 TI - Frequency of work-related respiratory symptoms in workers without asthma. AB - BACKGROUND: Clinicians are faced with subjects complaining of work-related respiratory symptoms (WRS) without any evidence of asthma. We sought to assess the prevalence of subjects with WRS without asthma in a cohort of workers referred for possible work-related asthma (WRA) as well as compare the characteristics and the work environment of subjects with WRS to subjects with WRA. METHODS: A prospective observational study of workers referred for possible WRA over a 1-year period. Detailed medical and occupational questionnaires were administered. Pulmonary function tests as well as specific-inhalation challenges were performed. RESULTS: One hundred twenty workers were investigated. Fifty-one had WRA while 69 had WRS. The type and the severity of the respiratory symptoms were similar in both groups, except for wheezing which was more frequently reported in subjects with WRA (32 (62.7%)) than in subjects with WRS (16 (23.2%)) (P < 0.01). Both the workers with WRS and WRA were mainly employed in the manufacturing sector (64.7% (WRA) and 71% (WRS)). At the time of the first assessment 64.7% of subjects with WRA and 56.5% with WRS had left their workplace because of their bothersome respiratory symptoms. CONCLUSIONS: Subjects with WRS without asthma represent a large proportion of the subjects assessed in clinics specialized in the field of WRA. Like subjects with WRA, the population with WRS is likely to represent a significant medical burden. The similarity of the symptoms between the WRA and the WRS groups emphasizes the need to perform a thorough and objective investigation to diagnose WRA. PMID- 19308959 TI - Olfactory association learning and brain-derived neurotrophic factor in an animal model of early deprivation. AB - Animal models can serve to explore neural mechanisms underlying the effects of stressful early experiences on behaviors supporting attachment. Neonatal rats primarily use olfaction for attachment, and Brain-Derived Neurotrophic Factor (BDNF) may be a key transcription target in olfactory association learning. In this experiment, neonatal male and female rats were isolated individually for 3 hr daily in the first week of life while their dams were left with partial litters (Early Deprivation, ED) or remained undisturbed (Control). At 1 week of age, subjects were tested using a 2-day classical conditioning paradigm. The conditioned group (O/M) was exposed to a novel odor paired with a milk infusion. Three additional groups included an unpaired odor and milk exposure group (O/M unP), an odor exposure alone group (O/NM), and neither an odor nor a milk group (NO/NM). Learning the odor association, as revealed in a position preference for the novel odor, was accompanied by an increase in hippocampal BDNF in O/M subjects from undisturbed Control litters. BDNF levels were also positively related to degree of preference for the odor in the O/M Control group. ED subjects did not make the classically conditioned odor association and did not show an increase in hippocampal BDNF. ED increased BDNF levels in the olfactory bulb compared to Controls regardless of training group; individual levels were not correlated with performance because samples were pooled. These results suggest that changes in the transcription of BDNF may underlie some of the long term consequences of the early stress of maternal separation. PMID- 19308961 TI - A broad spectrum of developmental delay in a large cohort of prolidase deficiency patients demonstrates marked interfamilial and intrafamilial phenotypic variability. AB - Prolidase deficiency (PD) is a rare, pan-ethnic, autosomal recessive disease with a broad phenotypic spectrum. Seventeen causative mutations in the PEPD gene have been reported worldwide. The purpose of this study is to characterize, clinically and molecularly, 20 prolidase deficient patients of Arab Moslem and Druze origin from 10 kindreds residing in northern Israel. All PD patients manifested developmental delay and facial dysmorphism. Typical PD dermatological symptoms, splenomegaly, and recurrent respiratory infections presented in varying degrees. Two patients had systemic lupus erythematosus (SLE), and one a novel cystic fibrosis phenotype. Direct DNA sequencing revealed two novel missense mutations, A212P and L368R. In addition, a previously reported S202F mutation was detected in 17 patients from seven Druze and three Arab Moslem kindreds. Patients homozygous for the S202F mutation manifest considerable interfamilial and intrafamilial phenotypic variability. The high prevalence of this mutation among Arab Moslems and Druze residing in northern Israel, and the presence of an identical haplotype along 500,000 bp in patients and their parents, suggests a founder event tracing back to before the breakaway of the Druze from mainstream Moslem society. PMID- 19308960 TI - Linkage disequilibrium mapping of the chromosome 6q21-22.31 bipolar I disorder susceptibility locus. AB - We previously reported genome-wide significant evidence for linkage between chromosome 6q and bipolar I disorder (BPI) by performing a meta-analysis of original genotype data from 11 genome scan linkage studies. We now present follow up linkage disequilibrium mapping of the linked region utilizing 3,047 single nucleotide polymorphism (SNP) markers in a case-control sample (N = 530 cases, 534 controls) and family-based sample (N = 256 nuclear families, 1,301 individuals). The strongest single SNP result (rs6938431, P = 6.72 x 10(-5)) was observed in the case-control sample, near the solute carrier family 22, member 16 gene (SLC22A16). In a replication study, we genotyped 151 SNPs in an independent sample (N = 622 cases, 1,181 controls) and observed further evidence of association between variants at SLC22A16 and BPI. Although consistent evidence of association with any single variant was not seen across samples, SNP-wise and gene-based test results in the three samples provided convergent evidence for association with SLC22A16, a carnitine transporter, implicating this gene as a novel candidate for BPI risk. Further studies in larger samples are warranted to clarify which, if any, genes in the 6q region confer risk for bipolar disorder. PMID- 19308962 TI - Tin(II) doped anatase (TiO2) nanoparticles: a potential route to "greener" yellow pigments. AB - During our exploration of compounds in the Sn(II)-Ti(IV)-O system, we discovered that hydrolysis of titanium alkoxide solution in the presence of Sn(II) salts resulted in stable deep-yellow colored anatase nanoparticles. The samples were characterized by X-ray powder diffraction, electron microprobe, thermal analysis, transmission electron microscopy, and (119)Sn Mossbauer spectroscopy. Mossbauer data of the yellow colored samples showed the presence of both Sn(II) and Sn(IV) in a distorted environment as expected in the anatase structure. It is suggested that the cationic charge imbalance is compensated by oxygen vacancies and/or hydroxyl groups as evidenced by Mossbauer data which show two types of Sn(II) environments. When heated in air to 300 degrees C the samples changed color to completely white and (119)Sn Mossbauer data of these samples showed only the presence of Sn(IV). These observations indicate that the origin of the yellow color in our Sn doped anatase nanoparticles arises from filled Sn 5s states just above the O 2p band, thus decreasing the band gap. The Sn(II) doped anatase TiO(2) nanoparticles reported here can potentially lead to environmentally benign yellow pigments. The simplistic nature of the synthetic procedure could easily be adapted to large-scale industrial manufacture. PMID- 19308963 TI - Large entropic effect in flexible crystalline media for gas separation. AB - To develop the application of the adsorption ability of our flexible single crystal host [Cu(2)(bza)(4)(pyz)](n) (1) (bza = benzoate; pyz =pyrazine) possessing a 1D channel, we study the separation ability of a 1 packed column for various organic vapors and inorganic gases. A 1 packed column can detect various organic molecules with sharp signals although steric or nonpolar molecules give broad signals. Interestingly, 1 separates various organic mixtures even if the mixture contains nonpolar molecules. Comparing the separation properties with columns of other separation media, including zeolite, activated carbon, activated alumina, and silica gel, suggests that a 1 packed column separates various gaseous molecules under moderate conditions. Additionally, the eluted order of similar molecules, such as N(2)/O(2) and methanol/ethanol using the 1 packed column is different from the others (zeolite, activated carbon, activated alumina, and silica gel), which suggests a difference in the separation mechanism of 1. From GC measurements, the estimated changes in Gibbs free energy by gas adsorption, under diluted gas conditions, exhibits a large entropy dependence caused by regularity in the generated adsorption state, which enables the dynamic control of gas adsorption selectivity. Therefore, it is suggested that single crystal host 1, because of its flexibility, can separate various gases by adjusting its channel structure according to the features of the guest gaseous molecules. This generates active controllability of the adsorption potential in addition to the intrinsic adsorption interaction. PMID- 19308964 TI - A linkage search for joint panic disorder/bipolar genes. AB - There is comorbidity and a possible genetic connection between Bipolar disease (BP) and panic disorder (PD). Genes may exist that increase risk to both PD and BP. We explored this possibility using data from a linkage study of PD (120 multiplex families; 37 had > or =1 BP member). We calculated 2-point lodscores maximized over male and female recombination fractions by classifying individuals with PD and/or BP as affected (PD + BP). Additionally, to shed light on possible heterogeneity, we examine the pedigrees containing a bipolar member (BP+) separately from those that do not (BP-), using a Predivided-Sample Test (PST). Linkage evidence for common genes for PD + BP was obtained on chromosomes 2 (lodscore = 4.6) and chromosome 12 (lodscore = 3.6). These locations had already been implicated using a PD-only diagnosis, although at both locations this was larger when a joint PD + BP diagnosis was used. Examining the BP+ families and BP families separately indicates that both BP+ and BP- pedigrees are contributing to the peaks on chromosomes 2 and 12. However, the PST indicates different evidence of linkage is obtained from BP+ and BP- pedigrees on chromosome 13. Our findings are consistent with risk loci for the combined PD + BP phenotype on chromosomes 2 and 12. We also obtained evidence of heterogeneity on chromosome 13. The regions on chromosomes 12 and 13 identified here have previously been implicated as regions of interest for multiple psychiatric disorders, including BP. PMID- 19308965 TI - An association study of common variation at the MAPT locus with late-onset Alzheimer's disease. AB - The MAPT gene that encodes Tau is located on chromosome 17q21, in a region, which has evolved to form two major haplotypes, H1 and H2. There is strong evidence that the H1 haplotype, and a sub-haplotype (H1C), are overrepresented and associated with increased risk for the sporadic tauopathies, progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). Both PSP and CBD cases display Tau pathology similar to Late-Onset Alzheimer's Disease (LOAD). However, numerous association studies investigating the genetic involvement of MAPT in LOAD have generated conflicting results. Here we have used a large LOAD case-control sample to genotype SNPs that have been shown to define H1/H2 status and intra-H1 variability. Single marker association analyses found no evidence that any of the SNPs are associated with risk of LOAD. When gender and APOE4 status were taken into account we observed suggestive association for SNP rs242557 (P = 0.02). Stratification of the sample revealed association with rs242557 only in APOE4 positive individuals (P = 0.01 recessive model), however this result would not survive multiple correction. There was no significant difference in H1/H2 haplotype distribution between cases and controls. We also tested the association of specific sub-haplotypes on the H1 background and likewise results were negative. No effect was observed on disease age of onset for any of the markers studied. In summary, we find no evidence for allelic or haplotypic association, with SNPs in the MAPT gene and LOAD. SNP rs242557 is nominally significant in the APOE4 positive individuals. None of the SNPs studied modified AAO for LOAD. PMID- 19308966 TI - Importance of using highly pure internal standards for successful liquid chromatography/tandem mass spectrometric bioanalytical assays. AB - Internal standards (IS) with similar physicochemical properties to the analyte provide multiple advantages in liquid chromatography/tandem mass spectrometric (LC/MS/MS) bioanalytical methods such as: reduction of the analysis run time, improvement in the intra-injection reproducibility, impact reduction of matrix and ionization effects. However, it is important to evaluate the purity of the IS prior to their use. Indeed, a minor impurity in the IS may lead to an important issue during bioanalytical method development. Stable labelled internal standards are usually appropriate IS for bioanalysis. The use of oxycodone-D3, ursodiol-D5 and atovaquone-D4 as internal standards in three different bioanalytical methods was evaluated. During oxycodone, oxymorphone and noroxycodone simultaneous quantification method development, oxymorphone was identified as a contaminant in oxycodone-D3. Since the limit of quantification for oxymorphone was very low (10 pg/mL), the presence of an even low percentage of oxymorphone in oxycodone-D3 leads to the change of the stable labelled IS for an analogue, ethylmorphine. 23 Nordeoxycholic acid was preferred to ursodiol-D5 as internal standard for the ursodiol, tauroursodiol and glycoursodiol simultaneous quantification method. Indeed, more than 7% of ursodiol was identified in the ursodiol-D5 which could not be bypassed by decreasing the IS concentration without compromising the linearity. An atovaquone-D4 reference standard revealed the non-negligible presence of atovaquone-D5 to atovaquone-D8 that has a large impact on the method validation. Therefore, atovaquone-D4 was sent for recertification since its isotopic purity was found to be much less than the isotopic purity mentioned on its certificate of analysis. Consequently, during bioanalytical method development, the purity of the IS should be scrutinized. PMID- 19308967 TI - Fast liquid chromatography/multiple-stage mass spectrometry of coccidiostats. AB - Drugs that are used as medicines and also as growth promoters in veterinary care are considered as emerging environmental contaminants and in recent years concern about their potential risk to ecosystems and human health has risen. In this paper we used a method based on liquid chromatography/electrospray tandem mass spectrometry to analyze eight coccidiostatic compounds: diclazuril, dinitrocarbanilide (the main metabolite of nicarbazin), robenidine, lasalocid, monensin, salinomycin, maduramicin and nasarin. Multiple-stage mass spectrometry (MSn) based on the precursor ions [M+Na]+ (polyether ionophores), [M+H]+ (robenidine) and [M-H]- (diclazuril and dinitrocarbanilide) was used to study the fragmentation of these compounds. MSn data and genealogical relationships were used to propose a tentative assignment of the different fragment ions. Loss of water, decarboxylations, ketone beta-cleavages and rearrangement of cyclic ethers and amide groups were some of the fragmentations observed for these compounds. Liquid chromatography with a sub-2 microm particle size column was coupled to tandem mass spectrometry (LC/MS/MS) allowing the separation of these compounds in less than 7 min. Method detection limits ranging from 11 to 71 ng L(-1) and run to-run values in terms of relative standard deviation (RSD) (up to 12%) were obtained. PMID- 19308968 TI - Photoelectrochemical studies of gold electrodes chemically modified with single walled carbon nanotubes. AB - The preparation, characterization and unique photoelectrochemical characteristics of a gold electrode chemically modified with suitably derivatized single-walled carbon nanotubes (SWCNTs) are described. The SWCNT-chemically modified gold electrode (CNT-CME) is assembled by linking acid-shortened SWCNTs with carboxylic groups at the nanotube ends to a gold surface previously modified with a cysteamine monolayer. A persistent photocurrent is observed when the modified electrode is illuminated with visible light in the presence of an electroactive species (ascorbic acid, AsA). The influence of physical variables (applied potential, light intensity) and of chemical parameters (AsA concentration, pH) on the generated photocurrents is evaluated. With an appropriate applied potential this device behaves as a photodiode. PMID- 19308969 TI - Selective enhancement of carbon nanotube photoluminescence by resonant energy transfer. AB - Highly selective: Enhancement of the photoluminescence (PL) emission efficiency of selected chiral forms of semiconducting single-walled carbon nanotubes (SWCNTs) is presented (see figure). Excitation of Nile blue A in the presence of SWCNTs results in the quenching of its fluorescence. The energy is resonantly transferred to the (7,5) SWCNT whereas the (8,7) tube is not in resonance; hence, its PL remains unaffected.We report on a simple method for enhancing the efficiency of photoluminescence (PL) emission from selected chiral forms of semiconducting single-wall carbon nanotubes (SWCNTs). The method is based on the use of a fluorescent dye (Nile blue A) that shows the capability of resonant energy transfer on to nanotubes. The excitation of Nile blue A in the presence of SWCNTs results in the quenching of its fluorescence and the energy is resonantly transferred to certain chiral forms. The PL emission from these chiral forms shows a marked increase in efficiency signifying the occurrence of Forster type resonant energy transfer (FRET). Due to its simplicity, this procedure has widespread implications for the detection of carbon nanotubes as well as for their use as fluorophores in FRET-based in vivo and in vitro biological applications. PMID- 19308970 TI - Localized electrografting of vinylic monomers on a conducting substrate by means of an integrated electrochemical AFM probe. AB - Combinations of scanning electrochemical microscopy (SECM) with other scanning probe microscopy techniques, such as atomic force microscopy (AFM), show great promise for directing localized modification, which is of great interest for chemical, biochemical and technical applications. Herein, an atomic force scanning electrochemical microscope is used as a new electrochemical lithographic tool (L-AFM-SECM) to locally electrograft, with submicrometer resolution, a non conducting organic coating on a conducting substrate. PMID- 19308971 TI - Dispersed fluorescence and delayed ionization of jet-cooled 2-aminopurine: relaxation to a dark state causes weak fluorescence. AB - Dark state: The photophysics of 9H-2AP (see figure) depends strongly on the solvent and the temperature. In polar, aprotic solvents the fluorescence quantum yield is lower compared to protic ones. Analysis of the diffuse fluorescence spectra point to the existence of a dark singlet n,pi* state which is long-lived (1 microsecond). PMID- 19308972 TI - Role of excited states in low-energy electron (LEE) induced strand breaks in DNA model systems: influence of aqueous environment. AB - Low-energy electrons (LEEs) interact with a DNA model system to create a variety of excited states. In the gas phase, dissociative (sigma*) states are accessible by LEEs with energy <4 eV (see picture) and cause facile strand breaks through a dissociative electron attachment mechanism. However, under solvation these dissociative (sigma*) states are blue-shifted and are less accessible to LEEs. PMID- 19308973 TI - In situ plasmon-heating-induced generation of Au/TiO2 "hot spots" on colloidal crystals. AB - SERS you right: The plasmon heating of gold nanoshells is exploited to yield the local conversion of amorphous TiO(2) into anatase on the surface of polymeric colloidal crystals (see scheme). The resulting Au/TiO(2) spots are active substrates for surface-enhanced Raman spectroscopy and allow surface reactions and processes to be followed directly on-site. PMID- 19308974 TI - Influence of sodium cations of N3 dye on the photovoltaic performance and stability of dye-sensitized solar cells. AB - We report on the effect of substituting the two tetrabutyl ammonium counter ions of the standard N719 dye by sodium ions on the performance and stability of dye sensitized solar cells (DSCs). The disodium analogue of N719 in conjunction with a non-volatile electrolyte gives a conversion efficiency of 7.6% under standard global AM 1.5 sunlight. Devices maintain 99% of their initial performance after 1000 h under full sunlight aging at 50 degrees C. Electrochemical impedance spectroscopy and photovoltage transient decay studies reveal the evolution of the solar cell parameters during aging. Remarkably, upon aging a decrease in the rate of electron back reaction with the triiodide ions across the TiO(2)/electrolyte interface appears as well as enhanced electronic conduction in the TiO(2) film. PMID- 19308975 TI - Beta-substituted terthiophene [2]rotaxanes. AB - Two kinds of beta-substituted terthiophene [2]rotaxanes were synthesized using the host-guest pairs of the electron-deficient cyclophane cyclobis(paraquat-p phenylene) (CBPQT(4+)) and the electron-rich terthiophenes with diethyleneglycol chains at the beta-position. One is made from the alpha-position non-substituted terthiophene (3 T-beta-Rx) and the other is made from the alpha-dibromo substituted terthiophene (3 TBr-beta-Rx). The binding constants of the beta substituted terthiophene threads were confirmed to be smaller than that of the alpha-substituted terthiophene analogue. By UV/Vis absorption measurements, we confirmed the charge-transfer (CT) band in the visible region with an extinction coefficient of approximately 10(2) (M(-1) cm(-1)). Strong, but not quantitative, quenching of the terthiophene fluorescence was confirmed for the [2]rotaxanes. Although the beta-substituted terthiophene thread was electrochemically polymerizable, the [2]rotaxane 3 T-beta-Rx was not polymerizable. This result indicates that the interlocked CBPQT(4+) macrocycle effectively suppresses the electrochemical polymerization of the terthiophene unit because electrostatic repulsive and steric effects of CBPQT(4+) hinder the dimerization of the terthiophene radical cations. In the electrochemical measurement, we confirmed the shift of the first reduction peak towards less negative potential compared to free CBPQT(4+) and the splitting of the second reduction peak. These electrochemical behaviors are similar to those observed for the highly constrained [2]rotaxanes. The beta-substituted terthiophene [2]rotaxanes reported herein are important key compounds to prepare polythiophene polyrotaxanes. PMID- 19308976 TI - Enhancement of H2 adsorption in coordination framework materials by use of ligand curvature. AB - Solvothermal reaction of the ligands H(4)L(110) ((2,7 phenanthrenediyl)diisophthalic acid) and H(4)L(111) ([2,7-(9,10 dihydrophenanthrenediyl)]diisophthalic acid) with Cu(NO(3))(2) x 2.5 H(2)O in a slightly acidified mixture of DMF/1,4-dioxane/H(2)O afforded the solvated framework compounds [Cu(2)(L(110))(H(2)O)(2)](DMF)(7.5)(H(2)O)(5) (NOTT-110) and [Cu(2)(L(111))(H(2)O)(2)](DMF)(7.5)(H(2)O)(5) (NOTT-111), respectively. Crystal structure determinations confirmed that NOTT-110 and NOTT-111 have the same NbO framework structure, differing only at the 9 and 10 positions of the phenanthrene group. The BET surface areas for desolvated NOTT-110 and NOTT-111 were estimated to be 2960 and 2930 m(2) g(-1), respectively. Compared with their phenyl analogues, introduction of phenanthrene groups to these porous Cu(II)-carboxylate framework materials leads to an enhancement of H(2) adsorption. Thus, the H(2) isotherms for desolvated NOTT-110 and NOTT-111 confirm 2.64 and 2.56 wt % total H(2) uptake, respectively, at 1 bar and 78 K. NOTT-110 shows a high total H(2) storage capacity of 7.62 wt % at 55 bar and 77 K (8.5 wt % at saturation) with a total volumetric capacity of 46.8 g L(-1) at 55 bar and 77 K. PMID- 19308977 TI - Dynamic and structural properties of aqueous arsenic solutions. AB - Aqueous arsenic solutions are important in geochemistry, environmental chemistry, and biochemistry. Thus, studies of arsenic solutions that probe the structure of the hydrated arsenic ion and the behavior of water around As ions have wide ranging implications. The mechanisms for water dissociation around three arsenic ions (see picture) and structural details of the resulting aqueous complexes are presented. PMID- 19308978 TI - Nanostructured copper oxide on silica-zirconia mixed oxides by chemical implantation. AB - N,N-Dialkylcarbamato complexes of copper(II), [Cu(O(2)CNR(2))(2)] (R = All = allyl, C(3)H(5); iPr, CH(CH(3))(2)) were prepared with the aim of functionalizing silica and nanostructured silica-zirconia matrices. The mixed matrices for the grafting reactions were prepared by copolymerizing MAPTMS (methacryloxypropyltrimethoxysilane), the precursor for the silica matrix, with the zirconium tetranuclear derivative [Zr(4)O(2)(OMc)(12)] (OMc = methacrylate), the precursor for the zirconia nanoparticles. Suspension of the silica and silica zirconia matrices in a solution of the copper dialkylcarbamate led to the functionalization of the respective substrates. The composition, microstructure, morphology, and physicochemical nature of the copper species grafted on the matrices were investigated by FTIR, X-ray photoelectron spectroscopy (XPS), EPR, X-ray absorption spectroscopy (XAS), XRD, TEM, and dinitrogen adsorption. The effect of selected experimental parameters (the nature of the copper precursor and of the matrix, grafting time, thermal treatment) on the grafting reaction was investigated. The Cu/Si ratio is increased by increasing the grafting time and the ZrO(2)-SiO(2) matrix is more reactive to attack by the carbamato complexes than either prepared or commercial SiO(2). After functionalization of the matrix, thermal treatment yielded nanostructured copper(II) oxide clusters, average diameter 12-15 nm, uniformly supported on the silica and on the silica-zirconia matrices. PMID- 19308979 TI - Polymerase-catalysed incorporation of glucose nucleotides into a DNA duplex. AB - The enzymatic recognition of six-membered ring nucleoside triphosphates--in particular the 6'-triphosphates of (beta-D-glucopyranosyl)thymine, (2',3'-dideoxy beta-D-glucopyranosyl)thymine, (3',4'-dideoxy-beta-D-glucopyranosyl)thymine and (2',3'-dideoxy-beta-D-glucopyranosyl)adenine--was investigated. Despite the facts that the pyranose nucleic acids obtained by polymerisation of these monomers do not hybridise in solution with DNA and that the geometry of a DNA strand in a natural duplex differs from that of a pyranose nucleic acid, elongation of the DNA duplex with all four nucleotide analogues by Vent (exo(-)) polymerase was observed. Modelling experiments showed that hydrogen bonds are formed when 2',3' dideoxy-beta-homo-T building blocks or beta-D-gluco-T building blocks are incorporated opposite adenosine residues in the template but not when they are incorporated opposite thymine residues in the template. The model shows a near perfect alignment of a secondary hydroxy group at the end of the primer and the alpha-phosphate group of the incoming triphosphate. The results of these experiments provide new information on the role of the active site of the enzyme in the polymerisation reaction. PMID- 19308980 TI - Radical polymerization of vinyl acetate with bis(tetramethylheptadionato)cobalt(II): coexistence of three different mechanisms. AB - The complex [Co(II)(tmhd)(2)] (4; tmhd = 2,2,6,6-tetramethylhepta-3,5-dionato) has been investigated as a mediator for controlled radical polymerization of vinyl acetate (VAc) and compared with the analogue [Co(II)(acac)(2)] (1; acac = acetylacetonato). A relatively well controlled process occurs, after an induction time, with 2,2'-azobis(4-methoxyl-2,4-dimethylvaleronitrile) (V-70) as radical initiator at 30 degrees C. However, whereas the polymerization essentially stops after about six initiator half-lives in the presence of 1, it continues with a first-order rate law in the presence of 4. The successful simulation of the kinetic data shows that 4 operates simultaneously by associative (degenerative transfer, DT) and dissociative (organometallic radical polymerization, OMRP) mechanisms. The occurrence of OMRP was confirmed by an independent polymerization experiment starting from an isolated and purified [Co(tmhd)(2)](PVAc) macroinitiator. The polymer molecular weight evolves linearly with conversion in accordance with the expected values for one chain per Co atom when DT is the predominant mechanism and also during the pure OMRP process; however, observation of stagnating molecular weights at long reaction times with concomitant breakdown of the first-order rate law for monomer consumption indicates a competitive chain transfer process catalyzed by an increasing amount of Co(II). In the presence of external donors L (water, pyridine, triethylamine) the DT pathway is blocked and the OMRP pathway is accelerated, and polymerization with complex 4 is then about five times slower than with complex 1. The reversal of relative effective OMRP rate constants k(eff) (4>1 in the absence of external donors, 4<1 in their presence) is rationalized through competitive steric effects on Co(III)-C and Co(II)-L bond strengths. These propositions are supported by (1)H NMR studies and by DFT calculations. PMID- 19308981 TI - Ground- and excited-state tautomerization rates in porphycenes. AB - The rates of double hydrogen transfer in the ground and excited electronic states have been measured for porphycene and its derivatives by using a new method based on pump-probe polarization spectroscopy. Changing the strength of two intramolecular hydrogen bonds by altering the NHN distance leads to differences in the tautomerization rate exceeding three orders of magnitude. The reaction is considerably slower in the lowest electronically excited state. A correlation was found between the tautomerization rates and (1)H chemical shifts of the internal protons. PMID- 19308982 TI - On the track of novel triel-stabilised silylaminoiminoborenes. AB - Borenes and boranes: Silylaminoiminoborenes, such as depicted, were isolated after treatment of halogen triels with silylaminofluoroboranes. In addition, novel aryl- and silyl-substituted diaminofluoroboranes were also prepared in order to substantiate this reaction route.Reactions between the halogen triels AlClMe(2), AlBr(3), GaCl(3) and the silylaminofluoroboranes (Me(3)Si)(2)NB(F)NRSiMe(3) (R=SiMe(3), CMe(3)) afforded the silylaminoiminoborenes, which were isolated as the triel adducts, such as Me(3)Si(Cl(3)Ga)NBNRSiMe(3) (6). In order to extent this reaction path to other fluoroboranes, novel aryl- and silyl-substituted diaminofluoroboranes were synthesised. Because almost no open-chain diaminofluoroboranes had been structurally characterised previously, corresponding fluoroboranes containing no silyl groups were crystallised for purposes of comparison. In complex reactions with the arylsilylaminofluoroboranes [(2,6-(iPr)(2)C(6)H(3))(Me(3)Si)NB(F)NR(2), R=iPr, iBu], amine adducts of borenium salts such as [(iPr)(2)NH-->B(Bu)NH-2,6 (iPr)(2)C(6)H(3)](+)AlCl(4) (-) (13) were obtained. PMID- 19308983 TI - Detection of mismatched duplexes by synchronizing the pulse potential frequency with the dynamics of ferrocene/isoquinoline conjugate-connected DNA probes immobilized onto electrodes. AB - Cyclic voltammetry was performed at various scan rates for the duplexes from ferrocene/isoquinoline conjugate-connected DNA probes on gold electrodes. The relationship between the observed currents and the scan rates disclosed the enhanced bending elasticity of the mismatched duplexes compared with the fully matched duplexes. The difference of the dynamics was easily detected through the currents from the conjugate by adjusting the pulse potential frequency in square wave voltammetry. By using the present strategy, we succeeded in accurately detecting various naturally occurring single-nucleotide polymorphisms. PMID- 19308984 TI - Direct catalytic asymmetric synthesis of highly functionalized 2-methylchroman 2,4-diols via Barbas-list aldol reaction. AB - A practical and sustainable chemical process for the synthesis of highly substituted aldol<-->lactol products was achieved for the first time through the asymmetric Barbas-List aldol (BLA) reaction of 2-hydroxybenzaldehydes with acetone in the presence of a catalytic amount of trans-4-OH-L-proline (see scheme). PMID- 19308985 TI - A waterwheel-shaped meso-meso-linked porphyrin pentamer. AB - Wheels on water? A waterwheel-shaped porphyrin pentamer has been synthesized by palladium-catalyzed cross-coupling reactions. The key intermediate is a boronate porphyrin, in which four boronic ester groups are directly attached to the meso positions. PMID- 19308986 TI - Molar mass, radius of gyration and second virial coefficient from new static light scattering equations for dilute solutions: application to 21 (macro)molecules. AB - New static light scattering (SLS) equations for dilute binary solutions are derived. Contrarily to the usual SLS equations [Carr-Zimm (CZ)], the new equations have no need for the experimental absolute Rayleigh ratio of a reference liquid and solely rely on the ratio of scattered intensities of solutions and solvent. The new equations, which are based on polarizability equations, take into account the usual refractive index increment partial differential n/partial differential rho(2) complemented by the solvent specific polarizability and a term proportional to the slope of the solution density rho versus the solute mass concentration rho(2) (density increment). Then all the equations are applied to 21 (macro)molecules with a wide range of molar mass (0.2500 kg mol(-1)), for which the scattered intensity is no longer independent of the scattering angle, the new equations give the same value of the radius of gyration as the CZ equation and consistent values of the second virial coefficient. PMID- 19308987 TI - T1 glottic carcinoma involving the posterior commissure. AB - OBJECTIVES/HYPOTHESIS: The posterior commissure is an uncommon site of glottic carcinoma. The aim of the study was to compare the clinical and prognostic characteristics of glottic carcinoma of the posterior commissure and the vocal cords. STUDY DESIGN: Retrospective case control study. METHODS: The study group consisted of 40 patients aged 32 to 84 years (mean, 62.7 +/- 10.7) diagnosed with T1 glottic carcinoma involving the posterior commissure from 1960 to 2008. Data on clinical features and outcome were collected from the medical files and compared with the data for 42 patients aged 30 to 87 years (mean, 64.4 +/- 11.8) with T1 vocal cord carcinoma. RESULTS: : There were no significant differences in clinical characteristics between the groups except for the higher rate of smokers among the patients with vocal cord carcinoma (95% vs. 65%, P = .01). All patients were treated primarily with radiotherapy. Disease-free survival in the posterior commissure carcinoma group was 76.1% after 5 years and 72.3% after 10 years, and in the vocal cord carcinoma group, 95% after 5 and 10 years (P = .012). The risk of recurrence was higher when the tumor involved the posterior commissure (hazard ratio, 8.78; 95% CI, 1.12-68.5, P = .038). CONCLUSIONS: T1 glottic carcinoma involving the posterior commissure has a more aggressive biological behavior and a worse prognosis than T1 glottic carcinoma of the vocal cords. Smoking, which is an important pathogenetic factor in vocal cord carcinoma, plays a lesser role in posterior commissure carcinoma. Laryngoscope, 2009. PMID- 19308988 TI - Control mechanisms of differential translation of Hsp90 isoforms in 9L rat gliosarcoma cells. AB - Although the differential expression of heat shcok proteins, Hsp90alpha and Hsp90beta was extensively studied in many kinds of cells, the post transcriptional regulation of Hsp90 isoforms remains unclear. In control and GA treated rat gliosarcoma cells, it has been reported that the translational efficiency of hsp90alpha is higher than hsp90beta. In this study, we present evidences identifying the roles for leaky scanning and 5'-UTR sequence in translational regulation of Hsp90beta. The result of in vitro transcription and translation (IVTT) experiment showed that hsp90alpha exhibited higher translation efficiency than hsp90beta. Sequence analysis revealed that there is an out-of frame downstream AUG codon in hsp90beta gene. However, elimination of the downstream AUG by site-directly mutagenesis or introducing Kozak context sequence around the initiator AUG of hsp90beta open reading frame increased its translational efficiency, which indicated that leaky scanning might be a possible mechanism regulating hsp90beta. Furthermore, we also constructed a firefly luciferase reporter system to verify the effect of subsequent translation at the downstream out-of-frame AUG codon in 9L and A549 cells. Furthermore, it is believed that 5'-untranslated region (5'-UTR) also plays a significant role in translational control. We showed hsp90beta 5'-UTR gives rise to the reduction of the translation efficiency in IVTT experiment. Additionally, the reductive effect of hsp90beta 5'-UTR was further confirmed by luciferase reporter assay using truncated deletion analyses of 5'-UTR of hsp90beta. Our results support the hypothesis that ribosome leaky scanning mechanism and 5'-UTR sequence acts as negative regulators in hsp90beta mRNA. PMID- 19308989 TI - Daxx inhibits muscle differentiation by repressing E2A-mediated transcription. AB - The basic helix-loop-helix (HLH) E2A transcription factors bind to DNA as homodimers or as heterodimers formed with other basic HLH factors, activate gene expression, and promote differentiation of muscle, lymphoid, neuronal, and other cell types. These E2A functions can be inhibited by the Id proteins, HLH factors that sequester E2A in non-DNA binding dimers. Here we describe the direct interaction of E2A with Daxx, a broadly expressed non-HLH protein previously associated with apoptosis and transcriptional repression. Daxx inhibits E2A function, but not via an Id-like mechanism; rather, it recruits histone deacetylase activity to E2A-dependent promoters. Increased Daxx expression during muscle differentiation inhibits E2A-dependent expression of key myogenic genes and reduces myotube formation, while decreased Daxx expression promotes myotube formation. These results identify a new mechanism for limiting E2A activity and establish a link between Daxx-mediated gene regulation and control of cellular differentiation. PMID- 19308990 TI - The SUMO-E3 ligase PIAS3 targets pyruvate kinase M2. AB - Pyruvate kinase M2 (M2-PK) controls the rate-limiting step at the end of the glycolytic pathway in normal proliferating and tumor cells. Other functions of M2 PK in addition to its role in glycolysis are little understood. The aim of this study was to identify new cellular interaction partners of M2-PK in order to discover novel links between M2-PK and cellular functions. Here we show that the SUMO-E3 ligase protein PIAS3 (inhibitor of activated STAT3) physically interacts with M2-PK and its isoenzyme M1-PK. Moreover, we demonstrate that endogenous SUMO 1-M2-PK conjugates exist in mammalian cells. Furthermore, we show that transient expression of PIAS3 but not the RING domain mutant PIAS3 (C299S, H301A) is consistent with nuclear localization of M2-PK and PIAS3 and M2-PK partially co localize in the nucleus of these cells. This study suggests a link between PIAS3 and nuclear pyruvate kinase. PMID- 19308991 TI - Interpersonal interactions on online forums addressing eating concerns. AB - OBJECTIVE: Although some research suggests that online eating disorder forums promote "pro-eating-disorder" lifestyles and discourage recovery, other research suggests that such forums are an important source of interpersonal support. The current study extends this research by exploring the positive and negative behaviors encouraged on these forums and by comparing forum members' perceptions of support received from online and offline relationships to support received in relationships of age-matched controls. METHOD: In a survey of 60 forum members, we assessed information exchanged and support provided on eating disorder forums. Further, we assessed perceptions of social support for general and specific life concerns in this group of forum members as well as 64 age-matched university controls. RESULTS: Results show that both adaptive and maladaptive behaviors are encouraged on the forums, and that this encouragement has some influence on forum members trying out these behaviors. Overall, forum members reported receiving less support for their eating concerns as compared to their general life stressors, and they perceived less support for both their general concerns and eating concerns in their offline relationships as compared to their online forum relationships. Moreover, forum members reported receiving less support from their offline relationships as compared to support received in relationships by age matched controls. DISCUSSION: Forum members perceive less support in their important relationships than other peers do, and they seek out and participate in forums as a means of attaining greater social support. However, our research suggests that these forums also encourage dysregulated eating behaviors. Implications of online forum support and its impact on recovery are discussed further. PMID- 19308992 TI - Adaptation and evaluation of the Clinical Impairment Assessment to assess disordered eating related distress in an adolescent female ethnic Fijian population. AB - OBJECTIVE: Measurement of disease-related impairment and distress is central to diagnostic, therapeutic, and health policy considerations for eating disorders across diverse populations. This study evaluates psychometric properties of a translated and adapted version of the Clinical Impairment Assessment (CIA) in an ethnic Fijian population. METHOD: The adapted CIA was administered to ethnic Fijian adolescent schoolgirls (N = 215). We calculated Cronbach's alpha to assess the internal consistency, examined the association between indicators of eating disorder symptom severity and the CIA to assess construct and criterion validity, and compared the strength of relation between the CIA and measures of disordered eating versus with measures of generalized distress. RESULTS: The Fijian version of the CIA is feasible to administer as an investigator-based interview. It has excellent internal consistency (alpha = 0.93). Both construct and criterion validity were supported by the data, and regression models indicated that the CIA predicts eating disorder severity, even when controlling for generalized distress and psychopathology. DISCUSSION: The adapted CIA has excellent psychometric properties in this Fijian study population. Findings suggest that the CIA can be successfully adapted for use in a non-Western study population and that at least some associated distress and impairment transcends cultural differences. PMID- 19308993 TI - Salinophagia in anorexia nervosa: case reports. AB - We report two cases of pathological ingestion of salt as a feature of anorexia nervosa, which we have previously termed "salinophagia." Both cases were young women with anorexia nervosa of the purging subtype and of sufficient severity to necessitate inpatient treatment. In both instances, excessive quantities of salt were ingested in the context of treatment programs requiring nutritional rehabilitation, and motivated by a wish to despoil the food and render it distasteful, to rob its ingestion of any hedonic qualities. In one instance, this behavior pattern was imitated by other patients on the unit. Having first briefly described salinophagia in 1999, the first author has received considerable correspondence from other specialists suggesting that this is not an isolated phenomenon. The issues of phenomenology and treatment are further discussed. PMID- 19308994 TI - Perfectionism across stages of recovery from eating disorders. AB - OBJECTIVE: This study examined perfectionism in relation to recovery from eating disorders by comparing different conceptualizations of perfectionism across healthy controls and fully recovered, partially recovered, and active eating disorder cases, where full recovery was defined using physical, behavioral, and psychological indices. METHOD: Participants were primarily young adult females; 53 active eating disorder cases, 15 partially recovered cases, 20 fully recovered cases, and 67 healthy controls. Participants completed questionnaires assessing trait perfectionism, perfectionistic self-presentation style, and frequency of perfectionism cognitions, as well as a diagnostic interview to determine lifetime and current eating disorder diagnoses. RESULTS: A robust pattern emerged whereby the fully recovered individuals and healthy controls had similar levels of perfectionism that were significantly lower than the perfectionism levels of the partially recovered and active individuals with eating disorder, who were comparable to each other. DISCUSSION: These findings have implications for more clearly defining eating disorder recovery and for the role perfectionism may play in achieving full recovery. PMID- 19308995 TI - Validity and reliability of a Fijian translation and adaptation of the Eating Disorder Examination Questionnaire. AB - OBJECTIVE: Assessment of disordered eating has uncertain validity across culturally diverse populations. This study evaluated Eating Disorder Examination Questionnaire (EDE-Q) performance in an ethnic Fijian study population. METHOD: The EDE-Q was translated, adapted, and administered to school-going Fijian adolescent females (N = 523). A subsample (n = 81) completed it again within approximately 1 week. We assessed feasibility, internal consistency, and test retest reliability; evaluated construct validity through factor analysis and correlation with similar constructs; and examined the marginal utility of an additional question on traditional purgative use. RESULTS: Internal consistency reliability was adequate for the global scale and subscales (Cronbach's alpha = 0.66-0.91); retest reliability was adequate for both the languages (range of ICCs, 0.50-0.79, and of kappas, 0.46-0.81, excluding purging items). Construct validity was supported by significant correlations with measures of similar constructs. Factor analysis confirms multiple dimensions of eating disorder symptoms but suggests possible culture-specific variation in this population. The majority of respondents endorsing traditional purgative use (58%) did not endorse conventional EDE-Q items assessing purging. DISCUSSION: The EDE-Q is a valid measure of eating disorder pathology for ethnic Fijian adolescent females and measures a unitary underlying construct. PMID- 19308996 TI - Course and outcome of eating disorders in a primary care-based cohort. AB - OBJECTIVE: To study the course and outcome of patients with eating disorder detected in primary care. METHOD: General practitioners (GP's) provided information on the course and outcome of eating disorders in patients (n = 147) diagnosed with anorexia nervosa (AN) or bulimia nervosa (BN) identified during a Dutch nationwide primary care-based incidence study. The research team determined the outcome based on the data provided by the GP's. The mean duration of the follow-up was 4.8 years. RESULTS: About 57% of the patients initially diagnosed with AN and 61% of those diagnosed with BN were fully recovered. AN binge/purge subtype (ANBP) demonstrated the most extended median survival time of all diagnostic subgroups (the point at which half of the group has reached full recovery). Diagnostic crossover was low. A younger age at detection predicted recovery at outcome for AN and BN. One patient (AN) died. DISCUSSION: The results of this study on differences in outcome and low crossover support the diagnostic distinction between AN and BN in the DSM-IV. Early detection is of major importance for a favorable outcome. PMID- 19308997 TI - OxLDL upregulates caveolin-1 expression in macrophages: Role for caveolin-1 in the adhesion of oxLDL-treated macrophages to endothelium. AB - Caveolin-1, a principle component of caveolae, is present in several cell types known to play an important role in the development of atherosclerosis. In this study, its distribution and expression were studied in the arterial walls of hypercholesterolemic rabbits and apo-E-deficient mice and in oxidized low-density lipoprotein (oxLDL)-treated RAW264.7 macrophages. Immunohistochemical studies showed that staining for caveolin-1 expression was stronger in atherosclerotic lesions in hypercholesterolemic rabbits and apo-E-deficient mice compared to normal rabbits and mice and was closely associated with macrophages. OxLDL treatment increased caveolin-1 protein expression in RAW264.7 macrophages in a time- and dose-dependent manner. The increase in caveolin-1 expression was dependent on phosphorylation of the mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinase1/2 (ERK1/2), p38, and Jun N-terminal kinase (JNK) and the transcriptional activation and translocation of nuclear factor-?B (NF-kappaB). OxLDL also induced caveolin-1 mRNA expression and this effect was not seen in the presence of inhibitors for transcription or de novo protein synthesis. OxLDL increased the adhesion of RAW264.7 macrophages to endothelial cells via an increase in caveolin-1 expression, and the adhesion was reduced by the use of anti-caveolin-1 antibody or caveolin-1-specific shRNA. These results show that oxLDL increases caveolin-1 expression in macrophages through the MAPKs/NF-kappaB pathway. The caveolin-1 levels are closely associated with the adherence of monocytes/macrophages to endothelial cells and their accumulation within the arterial intima after hypercholesterolemia insult, resulting in the progression of atherosclerosis. PMID- 19308998 TI - Histone marks and chromatin remodelers on the regulation of neurogenin1 gene in RA induced neuronal differentiation of P19 cells. AB - Neurogenin1 is an important bHLH protein that plays crucial role in neurogenesis. We first show that the expression of ngn1 increases drastically in RA induced neuronal differentiation. During which, a three successive stages of the epigenetic changes surrounding the ngn1 gene are found correlated with a repression to activation of the gene in P19 cells. Recruiting of a repressive histone code H3K27me3 on the ngn1 gene is the dominant change in first repression stage, which is followed by the binding of the active codes of H3K9ac, H3K14ac, and the H3K4me3 in the second and third stages of RA treatment. Additionally, BRM but not BRG1 is specifically recruited to ngn1 gene at the third stage and is positively involved in the RA induced ngn1 expression. We propose that histone modifiers and chromatin remodelers are pivotal in the activation of the ngn1 gene in RA induced differentiation of P19 cells. PMID- 19308999 TI - Functional characterization of mesenchymal stem cells labeled with a novel PVP coated superparamagnetic iron oxide. AB - Magnetic resonance imaging of cells labeled with superparamagnetic iron oxide (SPIO) could be a valuable tool for tracking transplanted cells in living organisms. Human bone marrow-derived mesenchymal stem cells (hBMMSC) were labeled with a novel polyvinyl pyrrolidone (PVP)-coated SPIO. Prussian blue staining and electron microscopy revealed that almost all of the cells were efficiently labeled with PVP-SPIO nanoparticles. There were no signs of cytotoxicity, even at concentrations of up to 1600 microg Fe/ml of the nanoparticles, and the labeled cells were successfully visualized by in vitro cellular MRI. In addition, there was no significant alteration of the phenotype or the adipo/osteo/chondrogenic differentiation potential of the cells. This was in contrast to Feridex IV labeling that led to the inhibition of hBMMSC chondrogenesis. Following intramuscular injection in a rabbit hind limb ischemia model, the intercellular migration of the labeled cells toward the ablated site was clearly tracked through in vivo MRI. The localization of the transplanted cells observed by MRI correlated well with postmortem histological studies. These results demonstrate that the novel PVP-SPIO nanoparticles appear to be efficient MR contrast agents and may enable non-invasive in vivo tracking of stem cells in experimental and clinical settings during cell therapy. PMID- 19309000 TI - Targeted disruption of Bcl-xL in mouse keratinocytes inhibits both UVB- and chemically induced skin carcinogenesis. AB - Bcl-x(L) is one of several antiapoptotic proteins regulated by signal transducer and activator of transcription 3 (Stat3). We have recently shown that Stat3 is required for chemically induced and ultraviolet B (UVB)-induced skin carcinogenesis. In this study, the functional role of Bcl-x(L) in skin carcinogenesis was investigated using skin-specific Bcl-x(L)-deficient mice. In this model, Bcl-x(L) expression is disrupted in the basal compartment of mouse epidermis using the bovine keratin 5 (K5) promoter to drive expression of Cre recombinase (K5.Cre x Bcl-x(L) (fl/fl) mice). A significant increase in apoptosis induced by either UVB irradiation or 7,12-dimethylbenz[a]anthracene (DMBA) treatment was observed in the epidermis of Bcl-x(L)-deficient mice. Furthermore, an increase in apoptotic cells was noted in hair follicle keratinocytes, including those located in the bulge region. Cell proliferation was not affected by Bcl-x(L) deficiency following exposure to either UVB or 12-O tetradecanoylphorbol-13-acetate (TPA). Bcl-x(L)-deficient mice were more resistant than wild-type controls to skin tumor development with delayed onset and reduced number of tumors using either UVB or the DMBA/TPA two-stage regimen. Moreover, Bcl-2, Mcl-1, and survivin protein levels were increased in the epidermis of Bcl-x(L)-deficient mice in the absence of stimuli. Furthermore, levels of these antiapoptotic proteins were also high in skin tumors from Bcl x(L)-deficient mice that developed in response to either UVB or two-stage carcinogenesis protocols. Collectively, these studies demonstrate that Bcl-x(L) plays a role early in skin carcinogenesis through its anti-apoptotic functions to enhance survival of keratinocytes, including bulge region keratinocyte stem cells, following DNA damage. PMID- 19309001 TI - Isolation and characterization of farnesyl diphosphate synthase from the cotton boll weevil, Anthonomus grandis. AB - Farnesyl diphosphate synthase (FPPS) catalyzes the consecutive condensation of two molecules of isopentenyl diphosphate with dimethylallyl diphosphate to form farnesyl diphosphate (FPP). In insects, FPP is used for the synthesis of ubiquinones, dolicols, protein prenyl groups, and juvenile hormone. A full-length cDNA of FPPS was cloned from the cotton boll weevil, Anthonomus grandis (AgFPPS). AgFPPS cDNA consists of 1,835 nucleotides and encodes a protein of 438 amino acids. The deduced amino acid sequence has high similarity to previously isolated insect FPPSs and other known FPPSs. Recombinant AgFPPS expressed in E. coli converted labeled isopentenyl diphosphate in the presence of dimethylallyl diphosphate to FPP. Southern blot analysis indicated the presence of a single copy gene. Using molecular modeling, the three-dimensional structure of coleopteran FPPS was determined and compared to the X-ray crystal structure of avian FPPS. The alpha-helical fold is conserved in AgFPPS and the size of the active site cavity is consistent with the enzyme being a FPPS. PMID- 19309002 TI - Trade-off between immune stimulation and expression of storage protein genes. AB - Proteins stored in insect hemolymph may serve as a source of amino acids and energy for metabolism and development. The expression of the main storage proteins was assessed in bacterial-challenged honey bees using real-time (RT)-PCR and Western blot. After ensuring that the immune system had been activated by measuring the ensuing expression of the innate immune response genes, defensin-1 (def-1) and prophenoloxidase (proPO), we verified the expression of four genes encoding storage proteins. The levels of vitellogenin (vg) mRNA and of the respective protein were significantly lowered in bees injected with bacteria or water only (injury). An equivalent response was observed in orally-infected bees. The levels of apolipophorin II/I (apoLp-II/I) and hexamerin (hex 70a) mRNAs did not significantly change, but levels of Hex 70a protein subunit showed a substantial decay after bacterial challenge or injury. Infection also caused a strong reduction in the levels of apoLp-III transcripts. Our findings are consistent with a down-regulation of the expression and accumulation of storage proteins as a consequence of activation of the immune system, suggesting that this phenomenon represents a strategy to redirect resources to combat injury or infection. PMID- 19309003 TI - Expression and characterization of the PEBP homolog genes from Drosophila. AB - The phosphatidylethanolamine binding proteins (PEBPs) family is evolutionarily conserved and involved in different physiological phenomena. PEBPs were found in many species from bacteria to mammals. Despite numerous studies, PEBPs' biological function and mode of action remain elusive. Based on sequence homology, seven PEBP genes were detected in the Drosophila genome. Only one of them, the odorant binding protein (OBP), has been characterized. To date nothing is known concerning the expression pattern and biological roles of the six other PEBP genes. By RT-PCR and Western blot analysis, we examined expression of the PEBPs in different tissues and embryos. The 6 PEBPs were differentially expressed. Only one, CG10298, is specific of only one tissue: the testis. Additionally, by comparing in wild type and male-sterile mutants we show that CG10298 is present only during spermatid differentiation. Furthermore, by comparing structural parameters of the six PEBP proteins with those of human PEBP 1, we have established that PEBP CG10298 is most closely related to human PEBP. PMID- 19309004 TI - Group membership, group norms, empathy, and young children's intentions to aggress. AB - This study assessed the effect of ingroup norms and empathy on 6 and 9-year-old children's (N=161) attitudes and aggressive intentions toward outgroup members. Prior to an intergroup drawing competition against an outgroup, participants' empathy was measured, and they were randomly assigned to a simulated group with a norm of direct or indirect aggression, or no aggression norm. Results indicated participants' attitudes were less positive toward the outgroup vs. the ingroup, and that both direct and indirect aggressive intentions were displayed toward the outgroup. Most importantly, the ingroup was liked less when it had an aggression norm, and the participants' aggressive intentions were not enhanced by the group aggression norm. Empathy was a significant negative predictor of direct but not indirect aggression intentions. Implications for understanding the instigation and inhibition of children's aggression intentions are discussed. PMID- 19309005 TI - Unsanctioned aggression in rugby union: relationships among aggressiveness, anger, athletic identity, and professionalization. AB - Aggressive players who intentionally cause injury to their opponents are common in many sports, particularly collision sports such as Rugby Union. Although some acts of aggression fall within the rules (sanctioned), others do not (unsanctioned), with the latter tending to be less acceptable than the former. This study attempts to identify characteristics of players who are more likely to employ unsanctioned methods in order to injure an opponent. Male Rugby Union players completed questionnaires assessing aggressiveness, anger, past aggression, professionalization, and athletic identity. Players were assigned to one of two groups based on self-reported past unsanctioned aggression. Results indicated that demographic variables (e.g., age, playing position, or level of play) were not predictive of group membership. Measures of aggressiveness and professionalization were significant predictors; high scores on both indicated a greater probability of reporting the use of unsanctioned aggressive force for the sole purpose of causing injury or pain. In addition, players who had been taught how to execute aggressive illegal plays without detection were also more likely to report using excessive force to injure an opponent. Results provide further support that highly professionalized players may be more likely to use methods outside the constitutive rules of Rugby Union in order to intentionally injure their opponents. Results are discussed within the context of the increasing win at-all-cost attitude that is becoming more prevalent in sport and its implications for youth athletes. PMID- 19309006 TI - Chip-based enantioselective open-tubular capillary electrochromatography using bovine serum albumin-gold nanoparticle conjugates as the stationary phase. AB - In this study, chip-based enantioselective open-tubular CEC (OT-CEC) was developed employing BSA-gold nanoparticle (GNP) conjugates as a chiral stationary phase. An immobilization procedure was realized by prederivatization of the glass microchannel with (3-mercaptopropyl)-trimethoxysilane to provide thiol groups, which linked the BSA-GNP conjugates on the inner surface of the microchannels. Incorporation of GNPs into immobilization of BSA selectors greatly increased the BSA phase ratio and favored the BSA stationary phase generated sufficient EOF. Good resolutions of FITC-labeled ephedrine and norephedrine isomers were achieved with 36 mm effective separation channel length within 250 s. The constructed OT CEC microdevice exhibited good repeatabilities for run-to-run enantioseparations and kept an enantioselective lifetime of more than 1 month. The effects of pH values and concentrations of a running buffer on the selectivity and resolution of enantioseparations were investigated. PMID- 19309007 TI - Gold nanoparticle-enhanced chemiluminescence detection for CE. AB - Introducing gold nanoparticles (AuNPs) to the running buffer further improved the sensitivity of luminol-H(2)O(2) chemiluminescence (CL) detection for CE. This has led to the development of sensitive CE-CL assays of biomedically interesting compounds. Epinephrine and norfloxacin were taken as the model analytes. Epinephrine inhibited light emission from the AuNP-catalyzed CL while norfloxacin enhanced it. The CE-CL assays had detection limits of 6.9x10(-9) M for epinephrine and 7.3x10(-9) M for norfloxacin. It was noted that in the absence of AuNPs no CE-CL analytical signal was produced by epinephrine at 4.0x10(-5) M or norfloxacin at 1.5x10(-3) M under similar experimental conditions. Fluorescence spectroscopic measurements showed that although the fluorescence excitation/emission maxima remained the same, the fluorescence lifetime of luminol increased significantly in the presence of AuNPs (tau(2) increased from 8.49+/-0.12 to 9.18+/-0.047 ns in a two-exponential fit), indicating that the excited states of luminol molecules were stabilized through the interaction between luminol molecules and AuNPs. Finally, quantitation of epinephrine and norfloxacin in biological samples such as human urine by using the present AuNP enhanced CE-CL method was demonstrated. PMID- 19309008 TI - Monolithic silica xerogel capillary column for separations in capillary LC and pressurized CEC. AB - Monolithic capillary columns were prepared by the reaction of a mixture of potassium silicate solution and formamide. The surface of the monolith was coated with a thin film formed by a sol-gel method to increase the surface area of the monolith and simultaneously covered with C8 as stationary phase for reversed phase separation. The morphology of the monolithic column was investigated by SEM. Monolithic columns prepared in this manner showed high permeability and can be operated in capillary LC (CLC) mode at a pressure of 20 psi. PAHs were used to evaluate the separation performance of the stationary phase using CLC and pressurized CEC (pCEC). Efficiencies of 20 000 and 28 000 plates per meter for naphthalene were obtained in CLC and pCEC modes, respectively. Improvement in column efficiency and reduction in analysis time over CLC and improvement in operation facility and separation selectivity over CLC were found using pCEC mode. PMID- 19309011 TI - Differences in protein distribution between human plasma preparations, EDTA plasma and heparin-plasma, analyzed by non-denaturing micro-2-DE and MALDI-MS PMF. AB - The effects of plasma preparation methods on the status of human plasma proteins were analyzed by non-denaturing micro-2-DE followed with polypeptide assignment with MALDI-TOF MS and PMF. In order to facilitate the separation of high molecular-mass plasma proteins (up to ca. 2x10(3) kDa) in short separation time, agarose micro-IEF gels were employed. The comparisons of the 2-DE patterns between EDTA-plasma (1.0 mg EDTA/mL blood) and heparin-plasma (0.025 mg heparin/mL blood) revealed the differences in protein distribution around pI 5.4 6.5 and apparent molecular mass of ca. 1x10(3) kDa, which were mainly attributed to the presence of the complexes of complement C4b and C4b-binding protein in heparin-plasma and their absence in EDTA-plasma. The distribution of several spots around pI 5.0-5.6 and apparent molecular mass 1.2-1.5x10(2) kDa was also found to be different; the fragments of complement C3 and C4 were detected in heparin-plasma but not in EDTA-plasma. The 2-DE pattern of high-heparin-plasma (0.50 mg heparin/mL blood) showed pI changes of three plasma proteins, fibronectin, complement factor B, and pre-alpha-inhibitor when compared with that of heparin-plasma, suggesting the interactions between heparin and the proteins. These results demonstrated that subtle changes in plasma proteins, caused by the different plasma preparation procedures, can be analyzed by non-denaturing agarose-IEF/micro-2-DE followed by MALDI-MS and PMF. PMID- 19309012 TI - Performance of agarose IEF gels as the first dimension support for non-denaturing micro-2-DE in the separation of high-molecular-mass plasma proteins and protein complexes. AB - Agarose micro-column gels (1% w/v agarose, diameter 1.4 mm and length 35 mm) were prepared as the first-dimension IEF support for non-denaturing 2-DE and the performance was compared with that of polyacrylamide gels (4.2% T and 4.8% C, the same gel size) using a human plasma sample. Sorbitol was not added in the agarose IEF gels, since its presence not only delayed the focusing of the proteins but also deteriorated the protein resolution. The optimum IEF time of the agarose gels for separation of 2 microL plasma sample (ca. 120 microg proteins) was decided to be 46 min, which is much shorter than that of the polyacrylamide gels (75 min). MALDI-MS and PMF assignment of the spots on the micro-2-DE gels at apparent molecular mass above ca. 5x10(2) kDa and pI from 4 to 8 revealed that when polyacrylamide IEF gels were used, many of the high-molecular-mass proteins resided at the sample loading edge or in basic pI regions as smear bands. When agarose IEF gels were used, most of the high-molecular-mass proteins moved to more acidic pI positions and were better focused, and their apparent pI values matched well with those previously reported for purified proteins. These results demonstrated the advantages of agarose-IEF/2-DE for the separation of high molecular-mass proteins and protein complexes under non-denaturing conditions. PMID- 19309013 TI - 2-DE and MS analysis of key proteins in the adhesion of Lactobacillus plantarum, a first step toward early selection of probiotics based on bacterial biomarkers. AB - The identification of cell components involved in probiotic activities is a challenge in current probiotic research. In this work, a new approach based on proteomics as an analytical tool for the identification of characteristic protein profiles related to adhesion to mucin as a model probiotic property was used. Three Lactobacillus plantarum strains with different adhesion rates were used for proteomic analysis: L. plantarum WHE 92 (15.9%), L. plantarum 299 v (9.1%) and L. plantarum CECT 4185 (1.4%). Cell wall extracts were subjected to proteomic analysis of differential protein expression using 2-DE, tryptic digestion, chip LC-QTOF mass analysis and protein identification using database search. Several proteins, previously reported to be involved in bacterial adhesion: elongation factor EF-Tu, GroEL chaperonin, molecular chaperone DnaK and glyceraldehyde-3 phosphate dehydrogenase were found to be overexpressed in the cell wall proteome of the highly adhesive strain L. plantarum WHE 92. The overexpression of two spots containing GroES co-chaperonin in the most adhesive strain also suggested the involvement of this protein in the adhesion process. The association of proteomic profiles and proteins with particular probiotic properties opens the way for the use of such profiles and proteins as bacterial biomarkers for the properties of bacteria but probably also for their potential health effects. PMID- 19309014 TI - Centrifugal methods and devices for rapid in-gel digestion of proteins. AB - Modern proteomic research frequently relies upon separation of proteins in a polyacrylamide gel matrix followed by in-gel enzymatic digestion and extraction of peptides for subsequent analysis by MS. In this work, we propose a novel semi automated method of mechanical processing of gel bands by passing these bands through a specially designed centrifugal device termed a Gel Shredder prior to digestion and extraction of peptides. Such a device allows integrated washing, destaining and shredding of gel bands into uniform blocks of controlled size, approximately 150-300 microm, prior to the enzymatic digestion and extraction of peptides. Shredding into uniform blocks increases the surface area of the gel pieces and promotes improved gel rehydration, allowing improved diffusion of the proteolytic enzymes and solvent into the gel lattice. We demonstrate that the new method substantially reduces the time spent on tedious manual handling of gel bands, while minimizing the risk of sample contamination. The performance of the Gel Shredder has been compared with a conventional in-gel digestion protocol using several standard proteins and a complex proteomic sample in terms of relative quantitation by either MALDI-TOF/TOF or nanoLC-ESI IT-Fourier transformation ion cyclotron resonance MS. It is shown that significant time savings and improved peptide recovery can be obtained for many proteins using the Gel Shredder compared with the traditional in-gel digestion protocol. PMID- 19309015 TI - 2-D PAGE-based comparison of proteasome inhibitor bortezomib in sensitive and resistant mantle cell lymphoma. AB - Although gene expression following bortezomib treatment has been previously explored, direct effects of bortezomib-induced proteasome inhibition on protein level has not been analyzed so far. Using 2-D PAGE in five mantle cell lymphoma cell lines, we screened for cellular protein level alterations following treatment with 25 nM bortezomib for up to 4 h. Using MS, we identified 38 of the 41 most prominent reliably detected protein spots. Twenty-one were affected in all cell lines, whereas the remaining 20 protein spots were exclusively altered in sensitive cell lines. Western blot analysis was performed for 17 of the 38 identified proteins and 70.6% of the observed protein level alterations in 2-D gels was verified. All cell lines exhibited alterations of the cellular protein levels of heat shock-induced protein species (HSPA9, HSP7C, HSPA5, HSPD1), whereas sensitive cell lines also displayed altered cellular protein levels of energy metabolism (ATP5B, AK5, TPI1, ENO-1, ALDOC, GAPDH), RNA and transcriptional regulation (HNRPL, SFRS12) and cell division (NEBL, ACTB, SMC1A, C20orf23) as well as tumor suppressor genes (ENO-1, FH). These proteins clustered in a tight interaction network centered on the major cellular checkpoints TP53. The results were confirmed in primary mantle cell lymphoma, thus confirming the critical role of these candidate proteins of proteasome inhibition. PMID- 19309016 TI - Adapted liquid crystal display backlighting unit for densitometry of stained polyacrylamide electrophoresis gels. AB - Appropriate monochromatic illumination that is spatially uniform will ensure densitometry of stained polyacrylamide electrophoresis gels with high sensitivity and repeatability. The approach of using an array of LEDs as an illuminator has practical limitations. Here we describe an alternative method based on adapted liquid crystal display backlighting. By changing the fluorescent light source and using a region where light intensity is uniform, we demonstrate the ability to achieve highly sensitive and repeatable densitometry measurements of polyacrylamide electrophoresis gels with Coomassie-stained proteins. PMID- 19309017 TI - Molecular identity of skin and blood T-cell clones in cutaneous T-cell lymphoma patients as determined from the migration pattern of the T-cell receptor-gamma gene by capillary electrophoresis. AB - The presence of a dominant clonal T-cell population in skin lesions is an important clue in the diagnosis of cutaneous T-cell lymphoma (CTCL). However, it has never been determined whether dominant T-cell receptor (TCR) rearrangements identified in skin lesions and blood from CTCL patients, displaying strictly identical migration patterns by capillary electrophoresis, actually correspond to identical clones. As this information has potential clinical relevance, TCR-gamma (TCRG) gene-derived amplified fragments from dominant blood and skin T-cell clones featuring either identical or slightly different capillary electrophoresis migration patterns were analyzed in eleven CTCL patients. In nine patients with identical electrophoretic migration patterns, sequence analyses revealed the dominant skin and blood T-cell clones to be identical. In contrast, in two patients displaying slight migration differences between skin and blood samples, the TCRG sequences were distinct. Additionally, capillary electrophoresis appears more sensitive and accurate than heteroduplex analysis and in silico analysis of samples of different origins is possible a posteriori. These results demonstrate the efficacy of capillary electrophoresis in assessing molecular identity and discrepancy of dominant T-cell populations obtained from different tissues or at different times, facilitating diagnosis and follow-up. PMID- 19309018 TI - Genetic polymorphisms in the DRD2, DRD3, and SLC6A3 gene in elderly patients with delirium. AB - Dopamine excess appears to be critical in the final common pathway of delirium. The aim of this study was to investigate whether genetic polymorphisms in three dopamine-related genes (the dopamine receptor 2 (DRD2), dopamine receptor 3 (DRD3), and the dopamine transporter (SLC6A3) gene) were associated with delirium. Patients aged 65 years and older acutely admitted to the medical department or to the surgical department following hip fracture were included. Delirium was diagnosed by the Confusion Assessment Method. Sixteen single nucleotide polymorphisms (SNPs) and one variable number of tandem repeats in the SLC6A3 gene, nine SNPs in the DRD2 gene, and six SNPs in the DRD3 gene were genotyped. Fifty percent of the 115 surgical patients and 34% of the 605 medical patients experienced delirium. Delirious patients were older and had more frequently pre-existing functional and cognitive impairment (P < 0.001). After correction for multiple testing, one SNP in the SLC6A3 gene (rs393795) was associated with reduced risk of delirium (P = 0.032). Adjusted for age, cognitive impairment, and functional impairment, three SNPs in the DRD2 gene and seven SNPs in the SLC6A3 gene were associated with delirium; none of these associations was significant after correction for multiple testing. Variations in the SLC6A3 gene and possibly the DRD2 gene were associated with delirium. Although validation of these results is needed our results support a role for the dopamine transporter and dopamine receptor 2 in the pathogenesis of delirium. PMID- 19309019 TI - COMT val158met influence on electroconvulsive therapy response in major depression. AB - There is strong evidence for a genetic contribution to the pathogenesis of depression, with the functional catechol-O-methyltransferase (COMT) val158met polymorphism having been suggested as a potential susceptibility factor. In the present study, the effect of COMT val158met on response to electroconvulsive therapy (ECT) was analyzed in a sample of 104 Caucasian patients (f = 71, m = 33) with pharmacologically treatment-resistant Major Depression. The higher active COMT 158val allele was found to be associated with (1) higher pre-ECT severity of depression and (2) better treatment response to ECT particularly regarding the core symptoms of depression as well as sleep-related symptoms. These findings were restricted to the female subgroup of patients. In summary, the present study supports a potentially gender-specific significant impact of COMT gene variation on electroconvulsive therapy response, with COMT 158val risk allele carriers suffering from more severe, pharmacologically less efficiently treatable depression and thus possibly deriving greater benefit from ECT in the first place. PMID- 19309020 TI - Radiation from mobile phone systems: Is it perceived as a threat to people's health? AB - The aim of this study was to explore the prevalence, nature and determinants of concerns about mobile phone radiation. We used data from a 2006 telephone survey of 1004 people aged 15+ years in Denmark. Twenty-eight percent of the respondents were concerned about exposure to mobile phone radiation; radiation from masts was of concern to about 15%. In contrast, 82% were concerned about pollution. Nearly half of the respondents considered the mortality risk of 3G phones and masts to be of the same order of magnitude as being struck by lightning (0.1 fatalities per million people per year) while 7% thought it was equivalent to tobacco induced lung cancer (approximately 500 fatalities per million per year). Among women, concerns about mobile phone radiation were positively associated with educational attainment, perceived mobile phone mortality risk and concerns about unknown consequences of new technologies. More than two thirds of the respondents felt that they had received inadequate public information about the 3G system. The results of the study indicate that the majority of the population has little concern about mobile phone radiation while a small minority is very concerned. PMID- 19309021 TI - Protecting-group-controlled surface chemistry-organization and heat-induced coupling of 4,4'-di(tert-butoxycarbonylamino)biphenyl on metal surfaces. AB - Like pearls on a string, molecular building blocks have been preorganized and then interlinked on a surface (see STM images). In this way both the supramolecular self-assembly of the reactants as well as the subsequent thermal activation to release the protecting group are controlled. PMID- 19309022 TI - Biotin-avidin as a protease-responsive cap system for controlled guest release from colloidal mesoporous silica. AB - Tell your guests when it's time to go: When avidin caps are attached to biotinylated colloidal mesoporous silica, the four subunits of the protein avidin can each bind to a biotin moiety attached to the surface (see picture). The resulting material is a promising candidate for the design of smart detergents or drug-delivery systems. The caps can be opened to release guest molecules by controlled enzymatic hydrolysis of the protein. PMID- 19309023 TI - Selective isomerization of epoxides to allylic alcohols catalyzed by TiO2 supported gold nanoparticles. AB - ReacTiO(2)ns for rings: Gold nanoparticles supported on TiO(2) are used as a novel heterogeneous catalyst for the isomerization of epoxides to allylic alcohols by a concerted mechanism (see scheme). The reaction proceeds in high yields and the product selectivity is often remarkable. PMID- 19309024 TI - Solution-based direct growth of organic crystals on an active channel region for printable bottom-contact organic field-effect transistors. AB - The growth and self-organization of organic crystals between a source (S) and drain (D) electrode by a method based on the use of a micropipette and isothermal evaporation of the solvent in a two-liquid system led to the formation of organic crystal transistors (see polarized optical micrograph). The method is similar to ink-jet printing and should be suitable for the fabrication of low-cost and mass producible printed electronic devices. PMID- 19309025 TI - Novel concepts in atherogenesis: angiogenesis and hypoxia in atherosclerosis. AB - The clinical complications of atherosclerosis are caused by thrombus formation, which in turn results from rupture of an unstable atherosclerotic plaque. The formation of microvessels (angiogenesis) in an atherosclerotic plaque contributes to the development of plaques, increasing the risk of rupture. Microvessel content increases with human plaque progression and is likely stimulated by plaque hypoxia, reactive oxygen species and hypoxia-inducible factor (HIF) signalling. The presence of plaque hypoxia is primarily determined by plaque inflammation (increasing oxygen demand), while the contribution of plaque thickness (reducing oxygen supply) seems to be minor. Inflammation and hypoxia are almost interchangeable and both stimuli may initiate HIF-driven angiogenesis in atherosclerosis. Despite the scarcity of microvessels in animal models, atherogenesis is not limited in these models. This suggests that abundant plaque angiogenesis is not a requirement for atherogenesis and may be a physiological response to the pathophysiological state of the arterial wall. However, the destruction of the integrity of microvessel endothelium likely leads to intraplaque haemorrhage and plaques at increased risk for rupture. Although a causal relation between the compromised microvessel structure and atherogenesis or between angiogenic stimuli and plaque angiogenesis remains tentative, both plaque angiogenesis and plaque hypoxia represent novel targets for non-invasive imaging of plaques at risk for rupture, potentially permitting early diagnosis and/or risk prediction of patients with atherosclerosis in the near future. PMID- 19309026 TI - Spin-polarized structures and solid-state NMR spectroscopy of paramagnetic compounds. AB - On an atomic scale and with high sensitivity, solid-state NMR spectroscopy can provide information about the electronic spin density and coupling mechanisms in paramagnetic compounds. The picture shows how the hyperfine splitting collapses through relaxation. Insights into which compounds are suitable and which approximations have to be made are given. PMID- 19309027 TI - Supramolecular chiral transcription and recognition by mesoporous silica prepared by chiral imprinting of a helical micelle. AB - Let's twist again: The chirality of a helical propeller-like micelle has been memorized by functional groups on the mesopore surface of chiral mesoporous silicas. Such imprinted supramolecular chirality can be transcripted to poly(propiolic acid) sodium salt and tetraphenylporphine tetrasulfonic acid (see picture), and recognized by B-DNA. PMID- 19309028 TI - Devising boron reagents for orthogonal functionalization through Suzuki-Miyaura cross-coupling. AB - Seeing the sites: The Suzuki-Miyaura reaction of substrates containing multiple coupling sites has been performed in a directed manner through the reactivity modulation of the boron moiety (see scheme). Several other strategies are also discussed. PMID- 19309029 TI - Towards an integrated chemical circuit. AB - Everything on a chip: Recent developments in microfluidics enable the combination of droplet microfluidics with continuous-flow systems (see picture). This is a promising step towards the development of integrated, complex synthesis and analysis laboratories on chips. For example, a building-block principle could be used to integrate a multistep reaction with purification and analysis. PMID- 19309034 TI - Long-term treadmill exposure protects against age-related neurodegenerative change in the rat hippocampus. AB - The potential of exercise or environmental enrichment to prevent or reverse age related cognitive decline in rats has been widely investigated. The data suggest that the efficacy of these interventions as neuroprotectants may depend upon the duration and nature of the protocols and age of onset. Investigations of the mechanisms underlying these neuroprotective strategies indicate a potential role for the neurotrophin family of proteins, including nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). In this study, we have assessed the effects of 8 months of forced exercise, begun in middle-age, on the expression of long-term potentiation (LTP) and on spatial learning in the Morris water maze in aged Wistar rats. We also assessed these measures in a cage control group and in a group of rats exposed to the stationary treadmill for the same duration as the exercised rats. Our data confirm an age-related decline in expression of LTP and in spatial learning concomitant with decreased expression of NGF and BDNF mRNA in dentate gyrus (DG). The age-related impairments in both plasticity and growth factor expression were prevented in the long-term exercised group and, surprisingly, the treadmill control group. Given the extensive handling that the treadmill control group received and their regular exposure to an environment outside the home cage, this group can be considered to have experienced environmentally enriched conditions when compared with the cage control group. Significant correlations were observed between both learning and LTP and the expression of NGF and BDNF mRNA in the dentate gyrus. We conclude that decreased expression of NGF and BDNF in the dentate gyrus of aged rats is associated with impaired LTP and spatial learning. We suggest that the reversal of these age related impairments by enrichment and exercise may be linked with prevention of the age-related decline in expression of these growth factors and, furthermore, that enrichment is as efficacious as exercise in preventing this age-related decline. PMID- 19309036 TI - 5-HT(1A) and NMDA receptors interact in the rat medial septum and modulate hippocampal-dependent spatial learning. AB - Cholinergic and GABAergic neurons in the medial septum/vertical limb of the diagonal band of Broca (MS/vDB) projecting to the hippocampus, constitute the septohippocampal projection, which is important for hippocampal-dependent learning and memory. There is also evidence for an extrinsic as well as an intrinsic glutamatergic network within the MS/vDB. GABAergic and cholinergic septohippocampal neurons express the serotonergic 5-HT(1A) receptor and most likely also glutamatergic NMDA receptors. The aim of the present study was to examine whether septal 5-HT(1A) receptors are important for hippocampal-dependent long-term memory and whether these receptors interact with glutamatergic NMDA receptor transmission in a manner important for hippocampal-dependent spatial memory. Intraseptal infusion of the 5-HT(1A) receptor agonist (R)-8-OH-DPAT (1 or 4 microg/rat) did not affect spatial learning in the water maze task but impaired emotional memory in the passive avoidance task at the higher dose tested (4 microg/rat). While intraseptal administration of (R)-8-OH-DPAT (4 microg) combined with a subthreshold dose of the NMDA receptor antagonist D-AP5 (1 microg) only marginally affected spatial acquisition, it produced a profound impairment in spatial memory. In conclusion, septal 5-HT(1A) receptors appears to play a more prominent role in emotional than in spatial memory. Importantly, septal 5-HT(1A) and NMDA receptors appear to interact in a manner, which is particularly critical for the expression or retrieval of hippocampal-dependent long-term spatial memory. It is proposed that NMDA receptor hypofunction in the septal area may unmask a negative effect of 5-HT(1A) receptor activation on memory, which may be clinically relevant. PMID- 19309037 TI - Increased hippocampal neurogenesis in the progressive stage of Alzheimer's disease phenotype in an APP/PS1 double transgenic mouse model. AB - Alzheimer's disease (AD) is a progressive neurodegenerative disease associated with senile beta-amyloid (Abeta) plaques and cognitive decline. Neurogenesis in the adult hippocampus is implicated in regulating learning and memory, and is increased in human postmortem brain of AD patients. However, little is currently known about the changes of hippocampal neurogenesis in the progression of AD. As brain tissues from patients during the progression of AD are generally not available, an amyloid precursor protein (APP)/presenilin1 (PS1) double transgenic mouse model of AD was studied. Bromodeoxyuridine (BrdU) labeling supported by doublecortin staining was used to detect proliferating hippocampal cells in the mice. Compared with age-matched wild-type controls, 9-month-old transgenic mice with memory impairment and numerous brain Abeta deposits showed increased numbers of proliferating hippocampal cells. However, 3-month-old transgenic mice with normal memory and subtle brain Abeta deposits showed normal hippocampal proliferation. Double immunofluorescent labeling with BrdU and either NeuN or glial fibrillary acidic protein was conducted in mice at 10 months (28 days after the last BrdU injection) to determine the differentiation of proliferating cells. The number of hippocampal BrdU-positive cells and BrdU-positive cells differentiating into neurons (neurogenesis) in 10-month-old mice was greater in transgenic mice compared with age-matched controls, but the ratio of hippocampal BrdU-positive cells differentiating into neurons and astroglia was comparable. These results suggest hippocampal neurogenesis may increase during the progression of AD. Targeting this change in neurogenesis and understanding the underlying mechanism could lead to the development of a new treatment to control the progression of AD. PMID- 19309038 TI - Effects of neonatal stress on markers of synaptic plasticity in the hippocampus: implications for spatial memory. AB - Early stressful adverse situations may increase the vulnerability to cognitive deficits and psychiatric disorders, such as depression. Maternal separation (MS) has been used as an animal model to study changes in neurochemistry and behavior associated with exposure to early-life stress. This study investigated the effects of neonatal stress (MS) on the expression of synaptic plasticity markers in the hippocampus and a purported relationship to cognitive processes. Spatial learning (Morris water maze) significantly increased the expression of total levels of the neural cell adhesion molecule (NCAM), as well as its three major isoforms (NCAM-120, -140, and -180) both in the control and MS groups. Interestingly, these increases in NCAM expression after learning were lower in MS animals when compared with control rats. MS induced a significant decrease in total levels of NCAM, and specifically, in the NCAM-140 isoform expression. In the hippocampus of MS rats there was a significant decrease in brain-derived neurotrophic factor and synaptophysin mRNA densities. Cell proliferation, measured as BrdU-positive cells, was also decreased in the dentate gyrus of MS rats. Altogether these results suggest that MS can alter normal brain development, providing a potential mechanism by which early environmental stressors may influence vulnerability to show cognitive impairments later in life. PMID- 19309039 TI - Hippocampal volume and asymmetry in mild cognitive impairment and Alzheimer's disease: Meta-analyses of MRI studies. AB - Numerous studies have reported a smaller hippocampal volume in Alzheimer's disease (AD) patients than in aging controls. However, in mild cognitive impairment (MCI), the results are inconsistent. Moreover, the left-right asymmetry of the hippocampus receives less research attention. In this article, meta-analyses are designed to determine the extent of hippocampal atrophy in MCI and AD, and to evaluate the asymmetry pattern of the hippocampal volume in control, MCI, and AD groups. From 14 studies including 365 MCI patients and 382 controls, significant atrophy is found in both the left [Effect size (ES), 0.92; 95% confidence interval (CI), 0.72-1.11] and right (ES, 0.78; 95% CI, 0.57-0.98) hippocampus, which is lower than that in AD (ES, 1.60, 95% CI, 1.37-1.84, in left; ES, 1.52, 95% CI, 1.31-1.72, in right). Comparing with aging controls, the average volume reduction weighted by sample size is 12.9% and 11.1% in left and right hippocampus in MCI, and 24.2% and 23.1% in left and right hippocampus in AD, respectively. The findings show a bilateral hippocampal volume loss in MCI and the extent of atrophy is less than that in AD. By comparing the left and right hippocampal volume, a consistent left-less-than-right asymmetry pattern is found, but with different extents in control (ES, 0.39), MCI (ES, 0.56), and AD (ES, 0.30) group. PMID- 19309042 TI - Acute exercise-induced bilateral thigh compartment syndrome. AB - Acute compartment syndrome of the thigh is rare due to the space's ability to accommodate large volumes of fluid and, with the exception of the lateral septum, its thin compliant linings. This article describes a case of bilateral exercise induced severe compartment syndrome treated with anterior and posterior fasciotomies. A 29-year-old man was admitted to intensive care with myoglobinuria. His left thigh was evaluated 18 hours later for compartment syndrome. The patient reported that 14 hours prior to initial presentation, he had participated in a 1-hour session of vigorous basketball. He gradually developed bilateral moderately severe thigh pain and tea-colored urine. Physical examination revealed pain secondary to passive stretch of both knees at 20 degrees flexion, plus firm anterior and posterior compartments to palpation. A handheld pressure monitor revealed the following compartment pressures: left anterior 80 mm Hg; left posterior 75 mm Hg; right anterior 45 mm Hg; and right posterior 50 mm Hg. Bilateral emergent anterior and posterior compartment fasciotomies were performed. The patient developed a significant severe distal motor and sensory neurological deficit on the left side, which recovered to 3/5 motor strength and protective sensation. At 6-month follow-up, he ambulated with the assistance of a left ankle foot orthosis. Acute severe compartment syndrome can occur following vigorous exercise. We recommend fasciotomies after exercise induced acute compartment syndrome rather than initial observation because of the severity of morbidity associated with undertreated compartment syndrome. PMID- 19309040 TI - Decreased neuronal differentiation of newly generated cells underlies reduced hippocampal neurogenesis in chronic temporal lobe epilepsy. AB - Hippocampal neurogenesis declines substantially in chronic temporal lobe epilepsy (TLE). However, it is unclear whether this decline is linked to altered production of new cells and/or diminished survival and neuronal fate-choice decision of newly born cells. We quantified different components of hippocampal neurogenesis in rats exhibiting chronic TLE. Through intraperitoneal administration of 5'-bromodeoxyuridine (BrdU) for 12 days, we measured numbers of newly born cells in the subgranular zone-granule cell layer (SGZ-GCL) at 24 h and 2.5 months post-BrdU administration. Furthermore, the differentiation of newly added cells into neurons and glia was quantified via dual immunofluorescence for BrdU and various markers of neurons and glia. Addition of new cells to the SGZ GCL over 12 days was comparable between the chronically epileptic hippocampus and the age-matched intact hippocampus. Furthermore, comparison of BrdU+ cells measured at 24 h and 2.5 months post-BrdU administration revealed similar survival of newly born cells between the two groups. However, only 4-5% of newly born cells (i.e., BrdU+ cells) differentiated into neurons in the chronically epileptic hippocampus, in comparison to 73-80% of such cells exhibiting neuronal differentiation in the intact hippocampus. Moreover, differentiation of newly born cells into S-100beta+ astrocytes or NG2+ oligodendrocyte progenitors increased to approximately 79% in the chronically epileptic hippocampus from approximately 25% observed in the intact hippocampus. Interestingly, the extent of proliferation of astrocytes and microglia (identified through Ki-67 and S 100beta and Ki-67 and OX-42 dual immunofluorescence) in the SGZ-GCL was similar between the chronically epileptic hippocampus and the age-matched intact hippocampus, implying that the proliferation of neural stem/progenitor cells in the SGZ-GCL of the chronically epileptic hippocampus was not obscured by an increased division of glia. Thus, severely diminished DG neurogenesis in chronic TLE is not associated with either decreased production of new cells or reduced survival of newly born cells in the SGZ-GCL. Rather, it is linked to a dramatic decline in the neuronal fate-choice decision of newly generated cells. Overall, the differentiation of newly born cells turns mainly into glia with chronic TLE from predominantly neuronal differentiation seen in control conditions. PMID- 19309043 TI - Small cell extraskeletal osteosarcoma. AB - Extraskeletal osteosarcoma is a rare malignant mesenchymal neoplasm that accounts for <4% of all osteosarcomas and approximately 1.2% of all soft tissue sarcomas. Among the extraskeletal osteosarcomas, the small cell type is extremely rare. This article describes a 31-year-old man who had small cell extraskeletal osteosarcoma arising from the semimembranosus muscle. An incisional biopsy was performed and the histopathological findings showed many small cells and osteoid formation. The results were reported as a malignant small round cell tumor, consistent with an extraskeletal Ewing's sarcoma or primitive neuroectodermal tumor. Immunohistochemically, the tumor showed reactivity with antibodies against CD99 and neuron-specific enolase, but not with antibodies against S100 protein, CD138, alpha smooth muscle actin, chromogranin, Ki-67, leukocyte common antigen, epithelial membrane antigen, CD30, or desmin. The patient refused neoadjuvant chemotherapy. One week after an open biopsy, a wide marginal resection was performed. The final diagnosis was small cell extraskeletal osteosarcoma. Adjuvant chemotherapy was performed using doxorubicin, ifosfamide, and cisplatin together with a total of 60 Gy of radiation therapy. At 2-year follow-up, the functional Enneking score of the operated lower extremity was 28 points. We performed chest computed tomography, magnetic resonance imaging, and positron emission tomography-computed tomography. There were no regional recurrence and distant metastasis. Presently the patient is disease free. PMID- 19309044 TI - Extraskeletal myxoid chondrosarcoma of the perineum. AB - Extraskeletal myxoid chondrosarcoma is an aggressive tumor with significant metastatic potential. A 40-year-old man presented with an asymptomatic mass at the left ischial fossa. Magnetic resonance imaging showed a 9x6-cm mass with variable signal intensity and cystic formations. A computed tomography scan of the chest showed multiple pulmonary nodules. A computed tomography-guided core needle biopsy was consistent with low-grade extraskeletal myxoid chondrosarcoma. Despite the presence of lung metastases, wide resection of the primary tumor was performed. Wide resection included the perineal mass and the adjacent ischiopubic ramus. Postoperative recovery was uneventful. One month postoperatively, thoracotomy was performed, and approximately 20 pulmonary nodules were excised. Histological analysis of the excised specimens was consistent with extraskeletal myxoid chondrosarcoma metastases. Two years postoperatively, the patient is alive without evidence of local recurrence or distant disease. Extraskeletal myxoid chondrosarcoma is an unusual soft tissue sarcoma with ultrastructural and molecular features distinct from that of skeletal myxoid chondrosarcoma. The EWS CHN gene fusion is highly specific to extraskeletal myxoid chondrosarcoma; the gene fusion is positive in >or=75% of cases. Treatment should include wide excision for local tumor control. Resection of the lung metastases is feasible. Because of the prolonged survival of patients with extraskeletal myxoid chondrosarcoma, long-term follow-up is recommended for early detection of local recurrence or distant metastases. PMID- 19309045 TI - Postsurgical chondrolysis of the shoulder. AB - There are multiple reports in the literature of chondrolysis following arthroscopic shoulder surgery. Although the etiology of these cases is not known for certain, there has been speculation that radiofrequency devices, young patient age, instability surgery, intra-articular pain pumps, and type of anesthetic may be precipitating factors. This article describes a case of a 37 year-old law enforcement officer who injured both shoulders and ultimately underwent nearly identical bilateral procedures: arthroscopic superior labrum anteroposterior (SLAP) repair, Bankart repair, capsulorrhaphy, acromioplasty, and distal clavicle excision. Intra-articular pain catheters were placed following both procedures, but the right-sided catheter never functioned properly, as evidenced by continuous leakage outside of her body until it was removed. Subsequently she had an arthroscopic lysis of adhesions done for residual stiffness, in which the left humeral head and glenoid cavity were noted to be completely devoid of articular cartilage. Over the ensuing months, multiple cortisone injections, 5 viscosupplementation injections, physical therapy, and narcotics all failed to relieve her left shoulder pain. Radiographs showed significant left glenohumeral joint space narrowing and a normal-appearing joint space on the right. Our impression was postsurgical chondrolysis of the left shoulder. The patient has recently undergone humeral hemiarthroplasty with nonprosthetic glenoid arthroplasty. This case differs from others reported in the literature in that nearly identical bilateral procedures were performed by the same surgeon, yet chondrolysis only developed on the side that had a functioning postoperative pain catheter. PMID- 19309046 TI - Idiopathic chondrolysis treated with etanercept. AB - Idiopathic chondrolysis of the hip in children has been well documented in the literature. The insidious nature of the symptoms and lack of early radiographic findings and diagnostic testing often delay diagnosis. Children often report a stiff, painful hip and have an associated limp in the absence of trauma or constitutional symptoms. Despite these symptoms it remains a poorly understood diagnosis with no identifiable cause. Some have speculated an inflammatory cause, as this disease exhibits joint space narrowing, presumably due to enzymatic activity similar to juvenile rheumatoid arthritis. Despite case reports attempting traction, physical therapy, nonsteroidal anti-inflammatories, steroids, and even operative intervention, no current treatment regimen exists that offers proven appreciable benefit. We hypothesized the powerful anti inflammatory properties of etanercept would provide symptomatic and radiographic improvement of idiopathic chondrolysis of the hip. This article presents a case of an adolescent boy with a stiff, painful left hip that failed treatment with traction, physical therapy, naproxen, and methotrexate, prior to initiating etanercept. After 1 year of daily etanercept therapy, the patient's hip motion improved in all directions and his pain completely resolved. This novel therapeutic approach offered symptomatic relief and radiographic improvement, and may provide an effective treatment strategy for this difficult disease. PMID- 19309047 TI - Dumbbell-type spinal solitary fibrous tumor with paraplegia. AB - Solitary fibrous tumors are rare tumors that most commonly arise in the pleura. This article describes a case of a large dumbbell-shaped solitary fibrous tumor of the thoracic spine that was causing partial paraplegia. The patient was a 75 year-old woman who presented with swelling of the upper back and weakness of the lower extremities. Magnetic resonance imaging revealed a dumbbell-shaped tumor mass compressing the spinal cord, predominantly at the level of T8. The tumor extended transversely from the bilateral foramen of T8-T10 to the back muscles. The patient experienced increasing weakness of the lower extremities, so a T7-T11 laminectomy was performed and the tumor was excised piecemeal. Microscopically, the tumor was found to be composed of a proliferation of fibroblastic spindle cells. Immunohistochemically, it was strongly positive for CD34 but negative for S100. These findings were consistent with a solitary fibrous tumor. Three months postoperatively, thoracic radiographs showed progression of spinal kyphosis. Posterior fusion with instrumentation was performed. Three years postoperatively, there was no clinical evidence of tumor recurrence. Spinal solitary fibrous tumors are rare, particularly dumbbell-shaped tumors; to our knowledge, only 2 have been reported in the literature. The correct diagnosis is aided by noting the bland fibrous histology, strong CD34 immunostaining, and absence of S100 and cytokeratin positivity. Although solitary fibrous tumors of the spine are rare, they should be considered in the differential diagnosis of intraspinal lesions, particularly those occurring near pleura. PMID- 19309048 TI - Femoral artery intimal injury following open reduction of an acetabular fracture. AB - Vascular injury is a devastating complication of internal fixation of acetabular fractures. While lacerations of the large vessels may lead to massive bleeding, they are obvious and are addressed immediately during the surgery. We encountered a subtle case of intimal injury and thrombosis of the femoral artery. A 20-year old man presented with a fractured right acetabulum and right fourth, fifth, and sixth ribs with hemopneumothorax after a high-velocity motor vehicle accident. Twelve days after the injury, open reduction and internal fixation of the acetabular fracture from an ilioinguinal approach was performed. Postsurgically, examination showed no palpable femoral arterial pulse distal to the inguinal ligament. Exploration of the common femoral artery showed arterial thrombus, which was removed by a Fogarty catheter; however, the pulselessness persisted. Further exploration revealed intimal injury of the femoral artery. Replacement of the damaged segment by an interpositional venous graft was performed and peripheral pulses were reestablished. The patient recovered uneventfully. Extreme care should be taken to avoid vascular injury in pelvic fracture surgery. Meticulous surgical technique, particularly in the middle window, is vital to minimizing the risk of this complication. The insidious nature of the intimal arterial injury underlines the necessity of close intraoperative and postoperative monitoring of limb vascularity to detect such vascular lesions immediately and prevent this potentially disastrous complication. PMID- 19309049 TI - Fracture of a reverse total shoulder arthroplasty retentive liner. AB - Reverse shoulder arthroplasty may be indicated in shoulders that cannot be effectively managed by conventional reconstruction procedures. Reported complications include instability, infection, hematoma, scapular notching, and premature component loosening. To our knowledge, however, fracture of a humeral component polyethylene inlay has not been described. A 72-year-old man presented with unstable, painful shoulders after undergoing unsuccessful open repairs and acromioplasties at an outside institution for bilateral massive rotator cuff tears. We performed a left reversed shoulder arthroplasty with a 36-mm glenosphere and a +9-mm polyethylene liner. In the operating room under general anesthesia with chemical paralysis and an interscalene block, his shoulder was stable. The patient required revision with a metal 9-mm lateralizer following an atraumatic dislocation 4 weeks postoperatively. Four weeks following the revision, the patient again presented with a dislocated left shoulder without antecedent trauma. During this second revision, the +6-mm standard polyethylene liner was replaced with a +6-mm lateral retentive polyethylene liner. A third revision surgery was performed after the patient experienced 2 more dislocations with minor activity. Intraoperatively, a fracture of the previously placed +6-mm retentive polyethylene liner was noted. This article may represent a case of dramatic scapular impingement by the humeral components. More subtle forms of scapular notching are common and may suggest significant long-term problems. Many reverse shoulder prostheses are being refined or modified in an attempt to eliminate or reduce the scapular notching phenomenon. PMID- 19309050 TI - Isolated avulsion fracture of the extensor carpi radialis brevis insertion due to a boxer's injury. AB - Avulsion fractures involving the radial wrist extensor tendons are extremely rare. Only 8 avulsion fractures of the extensor carpi radialis longus and 7 avulsion fractures of the extensor carpi radialis brevis have been previously reported. We know of no case involving an isolated avulsion fracture of the base of the third metacarpal where the extensor carpi radialis brevis attachment to the fracture fragment was still completely intact. This article reports a patient with an isolated avulsion fracture of the extensor carpi radialis brevis insertion treated by open reduction and internal fixation. The avulsion fracture was the result of a poor punching technique. The patient healed uneventfully, and returned to full painless activity. The senior author (S.W.B.) of this article is a former Tae Kwon Doe instructor and includes some editorial comments on proper technique for punching to avoid injury. PMID- 19309051 TI - Arthrodesis of the knee using computer navigation in failed total knee arthroplasty. AB - Arthrodesis is used most commonly as a salvage procedure for failed total knee arthroplasty (TKA). For successful arthrodesis, a stable fusion technique and acceptable limb mechanical alignment are needed. Although the use of TKA intramedullary alignment rods may be helpful in terms of achieving an acceptable limb mechanical axis, fat embolism and intramedullary dissemination of an infection or reactivation of latent infection may occur in failed TKA cases with an active infection or a past history of infections. In this situation, computer assisted surgery allows precise cuts to be made in the frontal and sagittal planes without breaching medullary cavities. Navigated femoral and tibial bone resections were performed perpendicular to the coronal mechanical axis. Also, femoral resection was conducted at 0 degrees of flexion to the sagittal axis, and the tibial resection at 7 degrees of flexion to the sagittal axis. The arthrodesis was held in proper axial and rotational alignment. First, the tibia was placed against the femur, ensuring good bony apposition mediolaterally and anteroposteriorly. Second, the rotational alignment was also rechecked based on the information obtained from the navigation system, suggesting that the mechanical axis will be neutral if the rotational alignment is correct because the tibial slope was increased posteriorly. Knee arthrodesis was completed using the Ilizarov method. Postoperative radiographs revealed satisfactory alignment. We believe that computer navigation could be an alternative surgical option for arthrodesis for failed TKA secondary to intra-articular infection and could be used for failed TKA with extra-articular deformity. PMID- 19309052 TI - Early fracture of a bioabsorbable tibial interference screw after ACL reconstruction with subsequent chondral injury. AB - Graft fixation in anterior cruciate ligament (ACL) reconstruction is commonly performed with bioabsorbable devices. This article presents a case of a broken bioabsorbable tibial interference screw (Gentle Threads; Biomet, Warsaw, Indiana) that presented as an intra-articular loose body 4 months after ACL reconstruction with posterior tibialis tendon allograft. A 19-year-old man presented with symptoms of pain and catching for 1 week but reported no history of trauma. The broken screw tip was identified on magnetic resonance imaging examination, and the remaining screw appeared to be overinserted into the tibia. During arthroscopic removal, a 10-mm screw tip was found in the lateral gutter. The ACL graft was found to be well fixed, but small areas of chondral damage were found in the patellofemoral and medial compartment. The patient's symptoms resolved postoperatively. To our knowledge, this is the earliest report of a broken bioabsorbable interference screw and only the second report of subsequent chondral injury due to intra-articular migration. Although rare, late breakage and intra-articular migration of bioabsorbable interference screws should be considered during the postoperative evaluation of any patient with pain or mechanical symptoms, regardless of trauma. This case also supports the importance of both measurement of tibial tunnel length and inspection of the intercondylar notch following interference screw insertion. Orthopedic surgeons performing ACL reconstruction must be aware of this possible complication and its potential for devastating chondral injury. PMID- 19309053 TI - Meniscal repair of the knee. AB - Since King's research on knee degeneration, meniscal-sparing techniques and other significant advances have improved outcomes. The anatomy, classification, and biomechanics of menisci are reviewed. Although the majority of traumatic meniscal damage occurs to the medial meniscus during twisting-type injuries, modern surgeons should be prepared to deal with patients of all ages with varied symptoms and mechanisms of injury. Non-operative versus operative treatments depend on many variables including the location of tear, age, and knee stability. If surgery is indicated, open and various arthroscopic techniques are available. Post-operative management and rehabilitation should be considered vital parts to any successful meniscal repair. PMID- 19309054 TI - Arthroscopic technique for the evaluation and treatment of posterior shoulder instability. AB - Posterior instability is becoming an increasingly recognized problem with today's contact athletes. Although not as common as anterior Bankart lesions, posterior capsulolabral pathology can lead to significant instability and pain. Open posterior repair is difficult and results in the literature have been inconsistent. However, there is recent evidence to support arthroscopic repair with capsular placation as a promising solution. Our technique for arthroscopic posterior capsulolabral repair will be reviewed with emphasis on the key aspects for a successful outcome. PMID- 19309055 TI - The pharmacological management of skeletal-related events from metastatic tumors. AB - Bone is one of the most common sites of metastatic disease from cancer, affecting around 400,000 patients each year. Skeletal-related events (SRE) include hypercalcemia, fractures, spinal cord compression or severe bone pain and may occur as a complication of metastasis. Bisphosphonate therapy is crucial in the prevention and treatment of SREs; zoledronic acid and pamidronate have both been shown to significantly reduce the development of SREs. Other agents such as corticosteroids and analgesics are also used in symptomatic management. The use of these agents in conjunction with radiation and surgery can help to improve patient outcomes. PMID- 19309056 TI - Double V-shaped cancellous osteotomy of the distal radius for Kienbock's disease. AB - Treatments of Kienbock's disease range from conservative measures-usually immobilization during the acute phase-to such procedures as joint leveling by radial shortening or ulnar lengthening, intercarpal fusion, silastic arthroplasty, and vascularized bone grafts. Joint-leveling procedures have many complications such as nonunion, hardware problems, and pain. In this study, 34 patients with Kienbock's disease, stage 1 through 3B in Lichtman's classification, were treated with a new technique: a double V-shaped metaphyseal osteotomy of the distal radius and dorsal tilt of the distal radius. The patients were evaluated clinically regarding pain, which decreased in 88% of patients; range of motion, which improved in all patients; and grip strength, which improved by 90%. Progression of the disease was assessed radiologically by determining the carpal height ratio and the carpal ulnar distance ratio. There were no cases with nonunion of the radial osteotomy at follow-up. This procedure is an effective treatment method for Kienbock's disease in wrists that do not have degenerative changes in the adjacent carpal joints. PMID- 19309057 TI - Locker room-acquired MRSA. PMID- 19309058 TI - Percutaneous temporal wiring method for minimally invasive plate osteosynthesis of a distal tibial fracture. AB - Reduction of distal tibial fractures-especially fractures that include an oblique or spiral fracture component-during operative treatment with minimally invasive plate osteosynthesis is not always easy. Unexpected and unacceptable displacement can sometimes be seen after plate fixation. We designed an effective fracture reduction method involving a percutaneous temporal wiring technique for the reduction of distal tibial fractures, including fractures that contain oblique or spiral fracture components, during the minimally invasive plate osteosynthesis procedure. Our method can consistently accomplish satisfactory reduction of these fractures and easily maintain reduced fractures, fix the plate, reduce the level of radiation exposure during the operation. PMID- 19309059 TI - Augmentation of tendon-bone interface healing with low-intensity pulsed ultrasound. AB - The capacity of low-intensity pulsed ultrasound to accelerate the integration of grafted tendon-within-bone tunnel was evaluated. The extensor digitorum longus tendons of 20 New Zealand white rabbits were transplanted into bone tunnels in both proximal tibias. One side was chosen randomly to receive low-intensity pulsed ultrasound. The other side served as control. Six rabbits each were sacrificed at 2 and 3 weeks postoperatively for biomechanical testing, and 2 rabbits each were sacrificed at 1, 3, 6, and 12 weeks postoperatively for histological study. At 2 weeks postoperatively, the mean maximal tensile strength in the ultrasound-treated group (30.676 N) was significantly stronger (P<.05) than the control group (17.924 N). There was no significant difference of the maximal tensile strength (33.674 vs 33.340 N; P=.753) between the ultrasound treated and control groups at 3 weeks postoperatively. In the ultrasound-treated group, the histological study revealed the interface filled with denser granulation tissue and diffuse perpendicular anchored fibers in the 1- and 3-week specimens, and interface new bone formation in the 6- and 12-week specimens. The biomechanical and histological studies demonstrated that the low-intensity pulsed ultrasound enhanced the healing of grafted tendon-to-bone tunnel in the early stages. Our findings indicate that low-intensity pulsed ultrasound is a potentially promising mechanical tool to accelerate early tendon healing to bone tunnel. PMID- 19309060 TI - Augmentation of bone healing by specific frequency and amplitude compressive strains. AB - A considerable proportion of bone fractures show signs of delayed or impaired healing. A novel device that applies interfragmentary compressive forces to the fracture site was developed and evaluated for its effect on fracture healing. The midshafts of the right tibias of 16 rabbits were osteotomized, leaving a 2-mm gap. A 4-pin, double-bar external fixator was fixed to the tibias. Interfragmental compressive displacement of 0.5 mm with a frequency of 1 Hz was applied 15 minutes per day. Mechanical stimulation began 1 week postoperatively and continued for 2 and 3 weeks. Control animals received no stimulation. Callus quality and area were assessed by radiographs. Maximum force and total energy absorbed up to failure of the healed bones were determined using a standard 3 point bending test. Postmortem radiographic examinations showed that mean radiographic measures in both anteroposterior and mediolateral views was significantly larger in experimental groups than controls (P=.029). Mechanical measures of the healed bones demonstrated that mean values of maximum force and absorbed energy were significantly higher in the experimental groups than in the control groups (P=.029). Both radiographic and mechanical measures were significantly higher after 3 weeks compared to 2 weeks of receiving stimulation (P<.05). PMID- 19309061 TI - C1-C2 intra-articular screw fixation for atlantoaxial subluxation due to rheumatoid arthritis. AB - While various surgical procedures have been developed for the treatment of atlantoaxial subluxation due to rheumatoid arthritis, C1-C2 intra-articular screw fixation was developed to reduce intraoperative injuries to vertebral arteries. The purpose of this study was to report the therapeutic outcome of this procedure, which was followed for >2 years. Preoperative symptoms were alleviated in all patients. Only 1 patient with subaxial canal stenosis underwent additional laminoplasty during follow-up. All patients with class IIIA or milder neural deficit according to Ranawat's classification showed improvement to class I or II. There were no surgery-related complications or incidents, including injuries to vertebral arteries. Bone union was observed in all patients. No change was observed in the reduced atlas-dens interval during follow-up. The atlantoaxial angle was -6 degrees to 30 degrees (average, 19.4 degrees) at follow-up, and was >or=30 degrees after surgery (fusion in an overextended position) in only 1 patient. Although postoperative deterioration of cervical alignment was observed in 4 patients (18.2%), there was no additional surgery due to deterioration of cervical alignment. With no surgery-related complications or incidents, this procedure could be a safe and acceptable option for atlantoaxial subluxation due to rheumatoid arthritis. PMID- 19309062 TI - ACGME core competencies: where are we? AB - Beginning in July 2002, the Accreditation Council for Graduate Medical Education (ACGME) instructed all residency programs to require their residents to demonstrate competency in 6 core areas: patient care, interpersonal and communication skills, medical knowledge, professionalism, practice-based learning, and systems-based practice. The goal was to have objective markers of performance that would serve as a gauge to determine a program's accreditation. To determine the experiences of orthopedic residency programs with regard to the ACGME's core competencies, a national survey was administered to orthopedic program directors and selected orthopedic residents. Of those orthopedic programs that responded, most appeared to be complying with the ACGME requirements. Both directors and residents thought patient care and medical knowledge ranked most important, while practice-based learning and systems-based practice were assigned the lowest ranks. Barriers to implementation of the core competencies included low priority compared with clinical duties, lack of faculty or resident education, and lack of formal orthopedic core competencies. Residents and program directors agreed that their programs would benefit from a definition of each of the core competencies, including a greater commitment to the processes involved in surgical procedures. This study demonstrated a commitment to the core competencies by the programs that responded. The survey also suggested this commitment would be aided by improved definitions of some of the competencies for the orthopedic resident. PMID- 19309063 TI - Comparison of hip and knee arthroplasty outcomes at early and intermediate follow up. AB - A common perception among clinicians and patients is that recovery is similar following total hip arthroplasty (THA) and total knee arthroplasty (TKA). Improvement in the outcomes of 337 THAs and 256 TKAs implanted by the same surgeon between April 2003 and November 2005 were compared. Improvement was measured using changes in Oxford hip and knee scores measured preoperatively, at first follow-up, and 1 year postoperatively for each patient. Improvements between preoperative review and first follow-up and between preoperative review and 1-year postoperative follow-up were significantly greater for THA compared to TKA patients. Improvements between first postoperative follow-up and 1-year postoperative follow-up were not significantly different between THA and TKA patients. Although THA patients displayed a significantly worse score preoperatively, they displayed a significantly better score at both first follow up and 1-year postoperative follow-up. While both procedures improve postoperative pain and physical function, as measured by the Oxford score, improvements measured relative to preoperative levels were significantly smaller for TKA compared to THA patients. Despite recent advances in knee arthroplasty surgery, a significant proportion of TKA patients achieve relatively poor outcome scores postoperatively. This study shows that pain and function improve less and more slowly in the early and intermediate postoperative periods for knee compared to hip arthroplasty patients. PMID- 19309064 TI - Multi-modal, pre-emptive analgesia decreases the length of hospital stay following total joint arthroplasty. AB - Traditional treatment of pain following total joint arthroplasty involves postoperative oral narcotic medications and intravenous patient-controlled analgesia, both of which can result in significant postoperative morbidity. Multi modal analgesia involving >or=2 classes of drugs acting on different receptor types may be as effective as single-narcotic/patient-controlled analgesia with fewer analgesic-related side effects. In addition, administering analgesia prior to surgery (pre-emptive) may reduce postoperative pain intensity. The current study was designed to compare the impact of multi-modal pre-emptive analgesia versus patient-controlled analgesia on postoperative nausea, rehabilitation participation, and length of stay following total joint arthroplasty. A retrospective chart review and comparison was performed for patients undergoing total joint arthroplasty who received either postoperatively patient-controlled analgesia or pre-emptive analgesia (scheduled postoperative oxycodone and a COX-2 inhibitor). Length of hospital stay for the pre-emptive group averaged 2.74 vs 3.28 days for patient-controlled analgesia patients. The patient-controlled analgesia group consumed significantly more intravenous morphine (17.7 mg vs 7.2) and experienced a three-fold increase in nausea. In addition, the patient controlled analgesia group was twice as likely to miss therapy and nearly 2 times more likely to be discharged to an extended care facility. The use of pre-emptive oxycodone and a selective COX-2 inhibitor decreased postoperative narcotic requirements and increased participation in rehabilitation. In addition, patients receiving pre-emptive analgesics had a decreased hospital length of stay and reduced likelihood of discharge to a skilled nursing facility. These data support the continued study and use of pre-emptive multi-modal analgesia paradigms in this population. PMID- 19309065 TI - Percutaneous fixation of unstable proximal humeral fractures with cannulated screws. AB - Proximal humeral fractures treated at level I trauma centers are typically displaced, unstable, high-energy injuries associated with injuries to other extremities and closed head injuries. One method that can be used to treat 2-part proximal humeral fractures, if the patient meets certain criteria, is closed reduction and percutaneous cannulated screw fixation. We conducted a retrospective review of 20 consecutive patients treated with this technique, and complete follow-up data were available for 14. Thirteen achieved union and functional range of motion at a mean of 10.5 weeks. Three experienced complications: failure of fixation, infection, and heterotopic ossification. The technique of closed reduction and percutaneous cannulated screw fixation is safe and effective for proximal humeral fractures in carefully selected patients. PMID- 19309066 TI - Surgical treatment of distal tibia fractures: a comparison of medial and lateral plating. AB - The surgical method of repairing distal tibial fractures remains controversial. Open reduction and plating is a popular method that can result in good fixation and retention of the achieved position. The usual approach for open plating is anterior. Although it offers good exposure to the tibia, the medial plating is at high risk of wound problems and nonunions. Also, if fixation of the fibula is required, an additional incision must be made on the lateral side of the shin. Lateral plating using a single lateral approach for treating distal tibial and fibular fractures has been reported to have good results; however, most of these studies were small series. We retrospectively evaluated and compared the clinical outcomes of distal tibial fractures treated with medial plating using an anterior approach and lateral plating using a lateral approach. Eighty-eight patients with distal tibial fractures treated with medial or lateral plating were retrospectively reviewed. The 88 fractures were divided into 2 groups: the medial plating group used an anterior approach and included 49 patients and the lateral plating group used a lateral approach and included 39 patients. Both groups were similar with respect to injury mechanism, union rate, malunion rate, operative time, functional score, and range of ankle motion (P>or=.14). Both medial and lateral plating for treating distal tibial fractures achieved good functional outcomes with a low malunion rate; however, the lateral plating group had a lower complication rate (P=.047) and fewer hardware problems (P<.001). PMID- 19309067 TI - Retrograde intramedullary nailing of femoral diaphyseal fractures caused by low velocity gunshots. AB - The purpose of this study was to evaluate the results of retrograde intramedullary nailing of femoral diaphyseal fractures caused by low-velocity gunshots. Seventy-three patients (68 men and 5 women) with 74 femoral diaphyseal fractures caused by gunshots were treated with reamed, retrograde, statically locked intramedullary nailing. The main outcome measures were fracture union, the need for additional operations, shortening, angular deformity, postoperative knee range of motion, and complication rate. Twenty-nine patients sustained additional injuries. During initial treatment, 18 patients underwent further orthopedic and nonorthopedic elective procedures. Three patients had fasciotomies for compartment syndrome. At follow-up, 3 patients had shortening >10 mm (range, 12 18 mm) and 1 had angulation >10 degrees (13 degrees recurvatum). Four patients underwent surgeries after discharge. Complications included 1 case of heterotopic bone formation, 1 hypertrophic nonunion that went on to union after dynamization, 2 postoperative positive blood cultures, and 1 wound dehiscence. There were no cases of septic arthritis. Our data indicate that there is a low incidence of shortening, angular deformity, complication, and infection rates following treatment of femoral gunshot diaphyseal fractures with a retrograde nail. This study demonstrates that this is an acceptable alternative for the treatment of these injuries. PMID- 19309068 TI - Femoroacetabular impingement. PMID- 19309069 TI - The Sivash constrained acetabular cup. PMID- 19309070 TI - Radiologic case study: posteriorly displaced L2-L3 interbody cage: complication of lumbar interbody fusion. AB - Enhance your diagnostic skills with this "test yourself" monthly column, which features a radiograph and challenges you to make a diagnosis. PMID- 19309071 TI - Solution structure and thermodynamics of 2',5' RNA intercalation. AB - As a means to explore the influence of the nucleic acid backbone on the intercalative binding of ligands to DNA and RNA, we have determined the solution structure of a proflavine-bound 2',5'-linked octamer duplex with the sequence GCCGCGGC. This structure represents the first NMR structure of an intercalated RNA duplex, of either backbone structural isomer. By comparison with X-ray crystal structures, we have identified similarities and differences between intercalated 3',5' and 2',5'-linked RNA duplexes. First, the two forms of RNA have different sugar pucker geometries at the intercalated nucleotide steps, yet have the same interphosphate distances. Second, as in intercalated 3',5' RNA, the phosphate backbone angle zeta at the 2',5' RNA intercalation site prefers to be in the trans conformation, whereas unintercalated 2',5' and 3',5' RNA prefer the gauche conformation. These observations provide new insights regarding the transitions required for intercalation of a phosphodiester-ribose backbone and suggest a possible contribution of the backbone to the origin of the nearest neighbor exclusion principle. Thermodynamic studies presented for intercalation of both structural RNA isomers also reveal a surprising sensitivity of intercalator binding enthalpy and entropy to the details of RNA backbone structure. PMID- 19309072 TI - Mechanistic study of copper-catalyzed aerobic oxidative coupling of arylboronic esters and methanol: insights into an organometallic oxidase reaction. AB - Copper-catalyzed aerobic oxidative coupling of arylboronic acid derivatives and heteroatom nucleophiles is a highly useful method for the formation of aryl heteroatom bonds. Mechanistic studies reveal that this reaction proceeds via an "oxidase"-style mechanism. Kinetic and spectroscopic studies establish that transmetalation of the aryl group from boron to Cu(II) is the turnover-limiting step and reoxidation of the reduced catalyst by O(2) is rapid. Further mechanistic analysis implicates the involvement of an aryl-copper(III) intermediate that undergoes facile C-O bond formation. PMID- 19309073 TI - Finding antigen culprits in an occupational disease. PMID- 19309074 TI - Methyl syringate: a chemical marker of asphodel (Asphodelus microcarpus Salzm. et Viv.) monofloral honey. AB - During the liquid chromatographic study of the phenolic fraction of monofloral honeys was detected in the asphodel honey ( Asphodelus microcarpus Salzm. et Viv.) chromatogram a distinctive peak not detected in other monofloral honeys such as Arbutus unedo L., Hedysarum coronarium , Eucalyptus spp., and Galactites tomentosa . After thin layer chromatography (TLC) purification and characterization by NMR and LC-MS/MS, the compound was identified as methyl syringate (MSYR) and confirmed against an original standard. Levels of MSYR were measured in honeys of 2005, 2006, and 2007 by HPLC-DAD analysis. Level determination of MSYR was repeated in 2008 for 2006 and 2007 honeys to evaluate chemical stability of this phenolic compound. Levels of MSYR measured 1 year after the sampling did not show significant statistical differences (p < 0.05). The stability of MSYR was also confirmed by 12 asphodel honey samples collected in 2005 that showed amounts of methyl syringate comparable with those found in fresh honey. For the evaluation of MSYR origin, samples of nectars were collected from flowers and the content of MSYR was measured. Levels of MSYR in honeys are originated from the nectar with an average contribution of the nectar to the honey of 80%. Melissopalinological analysis did not allow the attribution of the honey monofloral origin because levels of asphodel pollen were <6% for all analyzed samples. Previously reported levels of MSYR for robinia, rape, chestnut, clover, linden blossom, dandelion, sunflower, thyme, manuka, and fir honeys were <5 mg/kg. For this reason, a minimum level of 122.6 mg/kg for MSYR in asphodel honeys can be considered as a chemical marker and, unlike the melissopalynological analysis, can be used for the origin attribution and to evaluate the percent of asphodel nectar in the honey. PMID- 19309075 TI - Melanosis and quality changes of Pacific white shrimp (Litopenaeus vannamei) treated with catechin during iced storage. AB - Melanosis, microbiological, chemical, and physical changes of Pacific white shrimp (Litopenaeus vannamei) treated with catechin were monitored during iced storage of 10 days. Whole shrimp treated with catechin solution (0.05 or 0.1%) had retarded growth of psychrophilic bacteria and spoilage microorganisms including H(2)S-producing bacteria and enterobacteriaceae throughout storage in comparison with the control and those treated with 1.25% sodium metabisilfite (SMS) (P < 0.05). The lower increases in pH and total volatile base (TVB) content were obtained in the shrimp treated with catechin solution at both levels, compared with those of other samples (P < 0.05). Lipid oxidation, loss in freshness and melanosis were lowered by catechin treatment. In general, the efficacy of catechin in lowering melanosis and quality losses increased with increasing levels used. Additionally, catechin (0.01, 0.05, and 0.1% (w/v)) showed inhibitory activity toward polyphenoloxidase (PPO) of Pacific white shrimp in a dose-dependent manner. Therefore, catechin can be used as a promising melanosis inhibitor as well as an antimicrobial and an antioxidant in ice-stored shrimp. PMID- 19309076 TI - Soy protein isolate extruded with high moisture retains high nutritional quality. AB - High-moisture extrusion of soy protein isolate generates a highly palatable meat substitute. No systematic evaluation of the nutritional quality of soy processed in this manner has been performed. This study compared the growth rate of male and female mice fed diets containing soy protein isolate without extrusion or with high-moisture extrusion. Other measures of overall growth and animal health were examined. Minor differences in the parameters were observed. Overall, the extruded soy protein was equally nutritious as the unextruded soy protein for the animals. Hence, high-moisture extrusion may be considered a useful method to generate high-quality protein foods. A longer term feeding trial may be recommended to further define the nutritional adequacy of this protein. PMID- 19309077 TI - Investigation of the protein-protein aggregation of egg white proteins under pulsed electric fields. AB - Egg whites were exposed to pulsed electric fields (PEFs) to investigate the protein-protein aggregation. No insoluble protein aggregate was found when egg whites were exposed to PEFs at 25, 30, and 35 kV/cm for 400 micros. However, atomic force microscopy showed that the sizes of the protein particles increased. Native polyacrylamide gel electrophoresis (PAGE) demonstrated the existence of aggregates under PEFs at 35 kV/cm for 400 micros. Sodium dodecyl sulfate (SDS) PAGE in the presence and absence of 2-mercaptoethanol further indicated that sulfhydryl-disulfide interchange reactions occurred under PEFs. Differential scanning calorimetry scans showed 400 micros of PEF treatment at 35 kV/cm denatured 16.5% proteins. Insoluble egg white protein aggregates were induced by PEF (35 kV/cm, 800 micros) and heat (60 degrees C, 3.5 min) treatments. Disulfide bonds were the dominant binding forces in the formation of protein aggregates. However, the weakly noncovalent bonds play a much more important role in the protein aggregation forming in heat treatment (60 degrees C, 3.5 min) than that in PEF treatment (35 kV/cm, 800 micros). PMID- 19309078 TI - Controlling axial conformation in 2-arylpyridines and 1-arylisoquinolines: application to the asymmetric synthesis of QUINAP by dynamic thermodynamic resolution. AB - Unlike related biphenyl compounds, 2-arylpyridines and 1-arylisoquinolines can be induced to adopt preferentially one of two axial conformations by the presence of a sulfinyl substituent adjacent to the Ar-Ar bond. In the case of more substituted biaryls, the compounds are atropisomeric, and thermodynamic selectivities of about 4:1 may be attained on heating. In the case of less hindered compounds, conformer ratios of up to 20:1 may be achieved. Preferred conformations are deduced by comparison of experimental CD spectra with those derived from theory. The conformational preferences induced by the sulfoxides may be exploited in the asymmetric synthesis of atropisomers, including the ligand QUINAP, by dynamic resolution under thermodynamic control. PMID- 19309079 TI - In praise of toolmakers. PMID- 19309081 TI - Pyranonaphthoquinone lactones: a new class of AKT selective kinase inhibitors alkylate a regulatory loop cysteine. AB - The naturally occurring pyranonaphthoquinone (PNQ) antibiotic lactoquinomycin and related aglycones were found to be selective inhibitors of the serine-threonine kinase AKT. A set of synthetic PNQs were prepared and a minimum active feature set and preliminary SAR were determined. PNQ lactones inhibit the proliferation of human tumor cell lines containing constitutively activated AKT and show expected effects on cellular biomarkers. Biochemical data are presented supporting a proposed bioreductive alkylation mechanism of action. PMID- 19309080 TI - Hedgehog-Gli signaling pathway inhibitors as anticancer agents. PMID- 19309082 TI - Discovering potent inhibitors against the beta-hydroxyacyl-acyl carrier protein dehydratase (FabZ) of Helicobacter pylori: structure-based design, synthesis, bioassay, and crystal structure determination. AB - The discovery of HpFabZ inhibitors is now of special interest in the treatment of various gastric diseases. In this work, three series of derivatives (compounds 3, 4, and 5) were designed, synthesized, and their biological activities were investigated as potential HpFabZ inhibitors in a two phased manner. First, we designed and synthesized two series of derivatives (3a-r and 4a-u) and evaluated the enzyme-based assay against HpFabZ. Five compounds (3i-k, 3m, and 3q) showed potential inhibitory activity, with IC(50) values less than 2 muM. Second, a focused combinatorial library containing 280 molecules was designed employing the LD1.0 program. Twelve compounds (5a-l) were selected and synthesized. The activity of the most potent compound 5h (IC(50) = 0.86 muM) was 46 times higher than that of the hit 1. The high hit rate and the potency of the new HpFabZ inhibitors demonstrated the efficiency of the strategy for the focused library design and virtual screening. PMID- 19309083 TI - Dynamics of the reaction of O(3P) atoms with alkylthiol self-assembled monolayers. AB - We have studied the dynamics of the reactions of O((3)P) atoms with alkylthiol self-assembled monolayers (SAMs). Superthermal O((3)P) atoms, with a fairly broad distribution of laboratory-frame kinetic energies (mean = 16 kJ mol(-1), fwhm = 26 kJ mol(-1)), were generated by 355 nm photolysis of NO(2) introduced at a low pressure above the SAM surface. Nascent OH v' = 0 products were detected by laser induced fluorescence. SAMs of two different alkyl chain lengths, C(6) and C(18), were studied. The existence of SAM layers, and their robustness under our experimental conditions during the relevant measurement period, were confirmed by scanning-tunneling microscopy (STM). Reaction at the SAM surface was verified as the authentic source of the hydroxyl radicals using a perdeuterated C(6)D(13)-SAM sample. The OH appearance profiles as a function of photolysis-probe delay, and the rotational-state distributions at their peaks, were compared with those of liquid squalane (C(30)H(62), 2,6,10,15,19,23-hexamethyltetracosane). The reactivity of the SAMs and of squalane was found to be comparable. We conclude that the O((3)P) atoms must be able to access the more reactive secondary hydrogen atoms along the alkyl chains of the SAMs. We find no perceptible differences in reactivity or product energy disposal between the two SAM chain lengths. Both produce a substantial fraction of the OH with relatively high velocities, which must result from direct, impulsive reaction. There is also a slower component, with velocities consistent with a thermal, trapping-desorption mechanism. The proportion of this component appears to be lower for SAMs than for squalane. This would be compatible with the expected greater smoothness of the SAM surface at the molecular scale. We find little evidence for significant rotational excitation of the OH products, although the details of any correlation between translational and rotational energy release require further investigation. We compare our results with the limited available prior theoretical modeling of O((3)P) + SAM systems. PMID- 19309085 TI - Analysis of 7,8-dihydro-8-oxo-2'-deoxyguanosine in cellular DNA during oxidative stress. AB - Analysis of cellular 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dGuo) as a biomarker of oxidative DNA damage has been fraught with numerous methodological problems. This is primarily due to artifactual oxidation of dGuo that occurs during DNA isolation and hydrolysis. Therefore, it has become necessary to rely on using the comet assay, which is not necessarily specific for 8-oxo-dGuo. A highly specific and sensitive method based on immunoaffinity purification and stable isotope dilution liquid chromatography (LC)-multiple reaction monitoring (MRM)/mass spectrometry (MS) that avoids artifact formation has now been developed. Cellular DNA was isolated using cold DNAzol (a proprietary product that contains guanidine thiocyanate) instead of chaotropic- or phenol-based methodology. Chelex-treated buffers were used to prevent Fenton chemistry mediated generation of reactive oxygen species (ROS) and artifactual oxidation of DNA bases. Deferoxamine was also added to all buffers in order to complex any residual transition metal ions remaining after Chelex treatment. The LC-MRM/MS method was used to determine that the basal 8-oxo-dGuo level in DNA from human bronchoalveolar H358 cells was 2.2 +/- 0.4 8-oxo-dGuo/10(7) dGuo (mean +/- standard deviation) or 5.5 +/- 1.0 8-oxo-dGuo/10(8) nucleotides. Similar levels were observed in human lung adenocarcinoma A549 cells, mouse hepatoma Hepa-1c1c7 cells, and human HeLa cervical epithelial adenocarcinoma cells. These values are an order of magnitude lower than is typically reported for basal 8-oxo-dGuo levels in DNA as determined by other MS- or chromatography-based assays. H358 cells were treated with increasing concentrations of potassium bromate (KBrO3) as a positive control or with the methylating agent methyl methanesulfonate (MMS) as a negative control. A linear dose-response for 8-oxo-dGuo formation (r(2) = 0.962) was obtained with increasing concentrations of KBrO3 in the range of 0.05 mM to 2.50 mM. In contrast, no 8-oxo-dGuo was observed in H358 cell DNA after treatment with MMS. At low levels of oxidative DNA damage, there was an excellent correlation between a comet assay that measured DNA single strand breaks (SSBs) after treatment with human 8-oxo-guanine glycosylase-1 (hOGG1) when compared with 8-oxo-dGuo in the DNA as measured by the stable isotope dilution LC-MRM/MS method. Availability of the new LC-MRM/MS assay made it possible to show that the benzo[a]pyrene (B[a]P)-derived quinone, B[a]P-7,8-dione, could induce 8-oxo-dGuo formation in H358 cells. This most likely occurred through redox cycling between B[a]P-7,8-dione and B[a]P-7,8-catechol with concomitant generation of DNA damaging ROS. In keeping with this concept, inhibition of catechol-O-methyl transferase (COMT)-mediated detoxification of B[a]P-7,8-catechol with Ro 410961 caused increased 8-oxo-dGuo formation in the H358 cell DNA. PMID- 19309086 TI - Field-effect modulation of Seebeck coefficient in single PbSe nanowires. AB - In this Letter, we present a novel strategy to control the thermoelectric properties of individual PbSe nanowires. Using a field-effect gated device, we were able to tune the Seebeck coefficient of single PbSe nanowires from 64 to 193 microV x K(-1). This direct electrical field control of sigma and S suggests a powerful strategy for optimizing ZT in thermoelectric devices. These results represent the first demonstration of field-effect modulation of the thermoelectric figure of merit in a single semiconductor nanowire. This novel strategy for thermoelectric property modulation could prove especially important in optimizing the thermoelectric properties of semiconductors where reproducible doping is difficult to achieve. PMID- 19309087 TI - Syntheses and structures of phenyl-C81-butyric acid methyl esters (PCBMs) from M3N@C80. AB - Two new 6,6-open phenyl-C(81)-butyric acid methyl ester metallofulleroids, M(3)N@C(80)PCBM (M = Sc, Y), were synthesized by diazoalkane addition reactions and fully characterized. The results demonstrate that the reactive sites are the same for M(3)N@C(80) (M = Sc, Y) but dramatically different from that of C(60). PMID- 19309088 TI - Grand canonical free-energy calculations of protein-ligand binding. AB - The principles behind the computation of protein-ligand binding free energies by Monte Carlo simulation in the Grand Canonical Ensemble are described in detail, and two variations of the calculation are presented. The computation can be performed by bathing a protein binding site either with ligand images that interact with each other or with ligand images that pass through each other. The second method is theoretically more rigorous, but we show that both methods lead to the same result, and there are distinct numeric advantages to using ligand images that interact with each other. The Grand Canonical simulation provides gas phase binding free energies that can be converted to aqueous energies by generalized Born-surface area (GB/SA) solvation calculations to provide values that agree with experiment within +/-1.5 kcal/mol. However, the accuracy of these simple solvation calculations is a major limiting factor in the accuracy of the overall binding free-energy computation. The Grand Canonical simulation has several characteristics beneficial to free-energy calculations. One is that the number of parameters that must be set for the simulation is small and can be determined objectively, making the outcome more deterministic, with respect to choice of input conditions, as compared to perturbation methods. Second, the simulation is free from assumptions about the starting pose or nature of the binding site. A final benefit is that binding free energies are a direct outcome of the simulation, and little processing is required to determine them. PMID- 19309089 TI - Fluorescence control on panchromatic spectra via c-alkylation on arylated quinoxalines. AB - A coherent green fluorescence was obtained by butylation at the 2-position of panchromatic 2,3-diaryl-5,8-diarylquinoxalines (2) to give corresponding 2-butyl 2,3-diaryl-5,8-diaryl-1H-quinoxalines (3). Full color quinoxaline derivatives (2) were prepared from electronic modification at either the 2,3- or 5,8-positions at the peripheral ArX group or X group (X = -H, -OMe, -NPh(2), -NMe(2), -NMePh) of the quinoxalines. 2-Butylation converted one imine unit of the pyrazine ring to an amine group, which effectively altered the electron donor and acceptor functions to produce a coherent green fluorescence. PMID- 19309090 TI - DFT and ONIOM(DFT:MM) studies on Co-C bond cleavage and hydrogen transfer in B12 dependent methylmalonyl-CoA mutase. Stepwise or concerted mechanism? AB - The considerable protein effect on the homolytic Co-C bond cleavage to form the 5'-deoxyadenosyl (Ado) radical and cob(II)alamin and the subsequent hydrogen transfer from the methylmalonyl-CoA substrate to the Ado radical in the methylmalonyl-CoA mutase (MMCM) have been extensively studied by DFT and ONIOM(DFT/MM) methods. Several quantum models have been used to systematically study the protein effect. The calculations have shown that the Co-C bond dissociation energy is very much reduced in the protein, compared to that in the gas phase. The large protein effect can be decomposed into the cage effect, the effect of coenzyme geometrical distortion, and the protein MM effect. The largest contributor is the MM effect, which mainly consists of the interaction of the QM part of the coenzyme with the MM part of the coenzyme and the surrounding residues. In particular, Glu370 plays an important role in the Co-C bond cleavage process. These effects tremendously enhance the stability of the Co-C bond cleavage state in the protein. The initial Co-C bond cleavage and the subsequent hydrogen transfer were found to occur in a stepwise manner in the protein, although the concerted pathway for the Co-C bond cleavage coupled with the hydrogen transfer is more favored in the gas phase. The assumed concerted transition state in the protein has more deformation of the coenzyme and the substrate and has less interaction with the protein than the stepwise route. Key factors and residues in promoting the enzymatic reaction rate have been discussed in detail. PMID- 19309091 TI - Synthesis of angstrom-scale anatase titania atomic wires. AB - Using a nonhydrolytic solution approach, we demonstrate the bulk synthesis of extremely thin crystalline TiO2 atomic wires in the anatase phase with diameters reaching the atomic limit of a few angstroms (approximately 4-5 A). These nearly monodisperse, atomically thin, and soluble TiO2 wires fill a most important size gap in nanowire fabrication. Preliminary results on photocatalytic activity of the atomic wires are also presented on degradation of methylene blue under visible light. These atomic wires are expected to promote exchanges between theory and experiments in fundamental studies of a one-dimensional (1D) system and provide unique building blocks to construct high-performance devices. PMID- 19309092 TI - Microscopic theory of light-induced deformation in amorphous side-chain azobenzene polymers. AB - We propose a microscopic theory of light-induced deformation of side-chain azobenzene polymers taking into account the internal structure of polymer chains. Our theory is based on the fact that interaction of chromophores with the polarized light leads to the orientation anisotropy of azobenzene macromolecules which is accompanied by the appearance of mechanical stress. It is the first microscopic theory which provides the value of the light-induced stress larger than the yield stress. This result explains a possibility for the inscription of surface relief gratings in glassy side-chain azobenzene polymers. For some chemical architectures, elongation of a sample demonstrates a nonmonotonic behavior with the light intensity and can change its sign (a stretched sample starts to be uniaxially compressed), in agreement with experiments. Using a viscoplastic approach, we show that the irreversible strain of a sample, which remains after the light is switched off, decreases with increasing temperature and can disappear at certain temperature below the glass transition temperature. This theoretical prediction is also confirmed by recent experiments. PMID- 19309093 TI - Detailed atomistic molecular dynamics simulations of alpha-conotoxin AuIB in water. AB - We present results about the shape, size, structure, conformational stability, and hydrodynamics of alpha-conotoxin AuIB (a disulfide-rich peptide from the venom of Conus aulicus, recognized as a nicotinic acetylcholine antagonist with great pharmaceutical potential) from very long (0.5 mus) massively parallel molecular dynamics (MD) simulations in full atomistic detail. We extract coarse grained descriptors of protein shape (ellipsoid), and of translational and rotational mobilities, i.e., the basic components at the lowest hierarchical level in a multiscale modeling strategy. Structural analysis reveals the folded conformation and asymmetric shape to be strongly favored for conotoxin. In accordance with experimental findings, conformational stability is observed and found to be linked to the presence of the alpha-helix along the 15 residues and to the existence of the two disulfide bonds. We find rotational (D(r)) and translational (D(t)) diffusivities to be suitable descriptors of coarse-grained dynamics, i.e., of the hydrodynamic behavior, and obtain D(r) = 3.62 (+/-0.17) x 10(8) s(- 1) and D(t) = 1.08 (+/-0.4) x 10(- 10) m(2) s(- 1). We further compare the MD-computed coarse-grained descriptors with first principles theoretical predictions based on the extended Hess-Doi Fokker-Planck approach which relates particle shape and dimensions to diffusion coefficients. An excellent agreement between simulation data and analytical predictions is observed for both dynamical descriptors. This comparison strongly suggests that diffusivities of rigid biomolecules much larger than the alpha-conotoxin AuIB studied here can be obtained from the coarse-grained shape descriptor (ellipsoid) derived from relatively short MD simulations. PMID- 19309094 TI - Hydrogen bonding and pi-stacking: how reliable are force fields? A critical evaluation of force field descriptions of nonbonded interactions. AB - We have evaluated the performance of a set of widely used force fields by calculating the geometries and stabilization energies for a large collection of intermolecular complexes. These complexes are representative of a range of chemical and biological systems for which hydrogen bonding, electrostatic, and van der Waals interactions play important roles. Benchmark energies are taken from the high-level ab initio values in the JSCH-2005 and S22 data sets. All of the force fields underestimate stabilization resulting from hydrogen bonding, but the energetics of electrostatic and van der Waals interactions are described more accurately. OPLSAA gave a mean unsigned error of 2 kcal mol(-1) for all 165 complexes studied, and outperforms DFT calculations employing very large basis sets for the S22 complexes. The magnitude of hydrogen bonding interactions are severely underestimated by all of the force fields tested, which contributes significantly to the overall mean error; if complexes which are predominantly bound by hydrogen bonding interactions are discounted, the mean unsigned error of OPLSAA is reduced to 1 kcal mol(-1). For added clarity, web-based interactive displays of the results have been developed which allow comparisons of force field and ab initio geometries to be performed and the structures viewed and rotated in three dimensions. PMID- 19309095 TI - Beyond photobleaching, laser illumination unbinds fluorescent proteins. AB - Confocal and two-photon fluorescence microscopy techniques using genetically encoded fluorescent probes are widely used in cell biology. Beyond the common problems of photobleaching and phototoxicity, we present evidence that photounbinding also has the potential to compromise such methods, especially in quantitative studies. We show that laser intensities within excitation regimes typical for imaging approaches such as as fluorescence recovery after photobleaching (FRAP), photolysis, or fluorescence correlation spectroscopy (FCS) experiments can cause the dissociation of antibodies from their ligands. Indeed, both one- and two-photon excitation of a fluorescent anti-GFP antibody caused its dissociation from immobilized GFP in vitro. Importantly, with two-photon excitation, the laser intensity threshold for photobleaching was the same as for photounbinding. By contrast, with single-photon excitation, we found a range of laser intensities where photobleaching can be separated from photounbinding. This photounbinding effect was visualized and measured by rebinding a second fluorescent anti-GFP (Green Fluorescent Protein) antibody, indicating that the GFP remained functional for reassociation following the photoinduced dissociation. Finally, we show that this unbinding effect occurs only when at least one binding partner carries a fluorescent label. Our results show that this photounbinding effect can readily remain masked or be misinterpreted as photobleaching, which can compromise the quantitative interpretation of binding studies made using fluorescence microscopy. PMID- 19309096 TI - Special effect of beta-cyclodextrin on the aggregation behavior of mixed cationic/anionic surfactant systems. AB - Controllable aggregate transitions are achieved in this work by adding due amounts of beta-cyclodextrin (beta-CD) to mixed cationic/anionic surfactant aqueous solutions. In contrast to its "aggregate breaking" effect in single surfactant systems, aggregate growth is observed in nonstoichiometrical mixed cationic/anionic surfactant systems upon addition of beta-CD. The aggregate growth typically undergoes a micellar elongation and a following micelle-to vesicle transition, which in turn greatly influences the viscosity and absorbance of the solutions. A possible mechanism of this beta-CD-induced aggregate growth is proposed. In mixed cationic/anionic surfactant systems, the surfactants strongly tend to reach electroneutral equilibrium in aggregates. In the present case, added beta-CD is found to greatly facilitate the equilibrium by transferring the "major" component (whose molar fraction>0.5) of a cationic/anionic surfactant mixture from the aggregates to beta-CD cavities. Consequently, the surfactants in the aggregates approach electroneutral mixing, in favor of low-curved aggregates such as vesicles. This work shows that beta-CD provides an additional degree of freedom to control microstructures and macroproperties for the whole class of mixed cationic/anionic surfactant systems. PMID- 19309097 TI - Observation and characterization of the CH3S(O)CH- and CH3S(O)CH- x H2O carbene anions by photoelectron imaging and photofragment spectroscopy. AB - We report the observation of the CH(3)S(O)CH(-) and CH(3)S(O)CH(-) x H(2)O carbene anions formed upon overall H(2)(+) abstraction from dimethyl sulfoxide by O(-). Photoelectron spectroscopy reveals singlet and triplet carbenes for the remaining neutral, with the singlet state assigned as the ground state. Although some formation of the distonic CH(2)S(O)CH(2)(-) radical anion is also expected, no conclusive evidence of the presence of this isomer is found. The photoelectron spectrum of HCSO(-) is also reported for the first time. Photofragmentation of CH(3)S(O)CH(-) with 532 nm light reveals two main types of anionic products: a dominant HCSO(-) fragment, resulting from methyl elimination, and a less intense SO(-) product. For the monohydrated anion, an additional SO(-) x H(2)O fragment is observed. Intriguingly, both the SO(-) x H(2)O and SO(-) products are produced with much higher yields in the fragmentation of CH(3)S(O)CH(-) x H(2)O, compared to the SO(-) yield from the dissociation of the bare CH(3)S(O)CH(-) anion. Two possible pathways are proposed as likely mechanisms for the SO(-)-based photoproducts, both involving a photoinduced intramolecular rearrangement and the formation of a C-C bond. PMID- 19309098 TI - Comparison of G3 and G4 theories for radical addition and abstraction reactions. AB - A test set of 21 radical addition and 28 hydrogen abstraction reactions has been studied at the W1, G4, G3X, G3X(MP2), and G3X(MP2)-RAD levels of theory with a view to establishing whether the recently introduced G4 theory offers improved performance over the G3 methods. All methods tested approximated the benchmark W1 values to within a mean absolute deviation (MAD) of 4 kJ mol(-1) or less, although maximum absolute deviations for problematic reactions (such as radical addition to thiocarbonyl compounds) can be as much as 10 kJ mol(-1) for the G3 methods. The new noncanceling higher-level correction (HLC) term in G4 was found to be capable of mitigating these errors in radical addition, but it introduced a systematic error to the reaction energies of the abstraction reactions, and its format may therefore require re-examination. G3 methods were shown to offer "chemical accuracy" even for these problematic cases, provided they were corrected to the W1 level of theory via an ONIOM-based approach. PMID- 19309099 TI - An optical voltmeter for studying cetyltrimethylammonium interacting with fused silica/aqueous interfaces at high ionic strength. AB - Electrostatics and counterions play important roles in many supramolecular processes, including surfactant adsorption and aggregation at interfaces. Here, we assess their influence on how the common surfactant cetyltrimethylammonium (CTA) interacts with fused silica/aqueous interfaces by determining thermodynamic, kinetic, and electrostatic parameters for CTA adsorption across a range of NaCl concentrations (10-500 mM NaCl) using second harmonic generation (SHG). Using vibrational sum frequency generation (SFG), we demonstrate that vibrationally resonant contributions and nonresonant background contributions to the SFG signal intensity that depend on the interfacial potential can be quantified simultaneously during the adsorption process, which provides insight into the nonequilibrium dynamics of CTA adsorption. By analyzing the adsorption free energies as a function of interfacial potential at these four salt concentrations, the charge density per adsorbate is determined, indicating that CTA coadsorbs with counterions at a ratio of approximately 4 to 3 (i.e., 4 CTA(+) ions for every 3 Cl(-) ion). The chemical (i.e., non-Coulombic) portion of the free energy is found to dominate the overall free energy of adsorption, indicating that CTA adsorption at these ionic strengths is primarily driven by the favorable hydrophobic interactions between interdigitated surfactant hydrocarbon chains in the adsorbed aggregate. By applying Gouy-Chapman-Stern theory to our data, an average charge density of 2.8(3) x 10(13) charges/cm(2), which corresponds to 0.7 to 1.7 molecules/nm(2), was obtained for the four NaCl concentrations. PMID- 19309100 TI - Density functional theory in prediction of four stepwise protonation constants for nitrilotripropanoic acid (NTPA). AB - It has been demonstrated for the first time that prediction of several consecutive protonation constants for the highly and negatively charged molecules, such as nitrilotripropanoic acid (NTPA), is possible with acceptable accuracy when isodesmic reaction (IRn) methodology, instead of commonly employed thermodynamic cycle (TC), is employed. Four stepwise protonation constants of NTPA were computed (RB3LYP/6-311+G(d,p) level of theory employing PCM/UA0 solvation model) to within +/-1 log unit of experimental data with an average error in the protonation constant of about 0.5 log unit. This good agreement was achieved for minimum energy structures of NTPA (studied ligand) and iminodiacetic acid (reference molecule). Results obtained strongly support the view that full conformational analysis should be seen as prerequisite for computing protonation/dissociation constants from IRn and possibly also from TC. Methodology proposed here broadens up, in our opinion, a scope of studying protonation constants computationally and opens up a new field of applications for poly charged ligands. TC did not work here at all as proton on N-atom was not preserved in gas-optimized structures; this proton always protonated available COO(-) group instead. PMID- 19309101 TI - Self-assembled monolayers of 2-Adamantanethiol on Au{111}: control of structure and displacement. AB - We have investigated the formation of 2-adamantanethiolate self-assembled monolayers on Au{111} and their displacement by n-dodecanethiol, using scanning tunneling microscopy, X-ray photoelectron spectroscopy, and infrared reflection absorption spectroscopy. Well-ordered 2-adamantanethiolate monolayers undergo rapid and significant molecular exchange upon exposure to n-dodecanethiol solutions, but their structures and intermolecular interactions template the growth of n-dodecanethiolate domains. Annealing 2-adamantanethiolate monolayers at 78 degrees C decreases the density of vacancy islands, while increasing the overall order and the average domain sizes in the films. This results in slower displacement by n-dodecanethiol molecules, as compared to unannealed monolayers. The secondary sulfur position on the adamantyl cage influences the lattice structure and exchange of 2-adamantanethiolate monolayers by alkanethiols. PMID- 19309102 TI - Nanoengineering of a biocompatible organogel by thermal processing. AB - The formation of most organogels requires the compatibility of both the gelator and solvent. It is very desirable if the rheological properties of a gel can be manipulated to achieve the desired performance. In this paper, a novel organogel was developed and its rheological properties and fiber network were engineered by controlling the thermal processing conditions. The gel was formed by the gelation of 12-hydroxystearic acid as a gelator in benzyl benzoate. It was observed that the degree of supercooling for gel formation has a significant effect on the rheological properties and fiber network structure. By increasing supercooling, the elasticity of the gel was enhanced, and the correlation length of the fibers was shortened, leading to the formation of denser fiber networks. The good biocompatibility of both the gelator and solvent makes this gel a promising vehicle for a variety of bioapplications such as controlled transdermal drug release and in vivo tissue repair. PMID- 19309103 TI - Peptide cyclization and cyclodimerization by Cu(I)-mediated azide-alkyne cycloaddition. AB - Head-to-tail cyclodimerization of resin-bound oligopeptides bearing azide and alkyne groups occurs readily by 1,3-dipolar cycloaddition upon treatment with Cu(I). The process was found to be independent of peptide sequence, sensitive to the proximity of the alkyne to the resin, sensitive to solvent composition, facile for alpha- and beta-peptides but not for gamma-peptides, and inhibited by the inclusion of tertiary amide linkages. Peptides shorter than hexamers were predominantly converted to cyclic monomers. Oligoglycine and oligo(beta-alanine) chains underwent oligomerization by 1,3-dipolar cycloaddition in the absence of a copper catalyst. These results suggest that cyclodimerization depends on the ability of the azido-alkyne peptide to form in-frame hydrogen bonds between chains in order to place the reacting groups in close proximity and lower the entropic penalty for dimerization. The properties of the resin and solvent are crucial, giving rise to a productive balance between swelling and interstrand H bonding. These findings allow for the design of optimal substrates for triazole forming ring closure and for the course of the reaction to be controlled by the choice of conditions. PMID- 19309104 TI - Structural studies of the ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate in dichloromethane using a combined DFT-NMR spectroscopic approach. AB - DFT methods in combination with NMR spectroscopy are used to investigate possible variants of the molecular structure of the ion pairs of the ionic liquid (IL) 1 ethyl-3-methylimidazolium tetrafluoroborate, [EMIM][BF(4)], in dichloromethane. According to the computations of the chemical shifts, experimental NMR spectra can be rationalized by an equilibrium between ca. 70-80% of structures with the anion positioned near to the C2 atom of the imidazolium ring and ca. 20-30% of structures with the anion close to the C5 and/or C4 atoms. The content of the latter structures, according to the computed Gibbs free energies, does not exceed 10%. Both the computations and the experimental NMR data suggest that the ratio of the two above-mentioned types of structures of the imidazolium-based ILs is influenced by the concentration/polarity of their dichloromethane solutions. PMID- 19309105 TI - Differential 12C-/13C-isotope dansylation labeling and fast liquid chromatography/mass spectrometry for absolute and relative quantification of the metabolome. AB - We report a new quantitative metabolome profiling technique based on differential (12)C-/(13)C-isotope dansylation labeling of metabolites, fast liquid chromatography (LC) separation and electrospray ionization Fourier-transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) detection. An isotope reagent, (13)C-dansyl chloride, can be readily synthesized. This reagent, along with (12)C-dansyl chloride, provides a simple and robust means of labeling metabolites containing primary amine, secondary amine, or phenolic hydroxyl group(s). It is shown that dansylation labeling offers 1-3 orders of magnitude ESI signal enhancement over the underivatized counterparts. Dansylation alters the chromatographic behaviors of polar and ionic metabolites normally not retainable on a reversed phase (RP) column to an extent that they can be retained and separated by RPLC with high efficiency. There is no isotopic effect on RPLC separation of the differential isotope labeled metabolites, and (12)C-/(13)C labeled isoforms of metabolites are coeluted and detected by MS for precise and accurate quantification and confident metabolite identification. It is demonstrated that, in the analysis of 20 amino acids, a linear response of over 2 orders of magnitude is achieved for relative metabolite quantification with an average relative standard deviation (RSD) of about 5.3% from replicate experiments. A dansylation standard compound library consisting of 121 known amines and phenols has been constructed and is proven to be useful for absolute metabolite quantification and MS-based metabolite identification in biological samples. As an example, the absolute concentrations of 93 metabolites, ranging from 30 nM to 2510 microM, can be determined from a pooled sample of human urines collected in 5 consecutive days labeled with (12)C-dansylation and spiked with the 121 (13)C-dansylated standards. Relative concentration variations of these metabolites in individual urine samples can also be monitored by mixing the (13)C dansylated pooled urine sample with the (12)C-dansylated individual sample. With a 12 min fast LC separation combined with FTICR MS, 672 metabolites were detected in a human urine sample with each metabolite peak having a signal-to-noise ratio of greater than 20; the identities of most of the metabolites remain to be determined. This work illustrates that dansylation labeling and fast LC/FTICR MS can be a powerful technique for quantitative profiling of at least 672 metabolites in urine samples in 12 min. PMID- 19309106 TI - Characterization of peptidoglycan in fem-deletion mutants of methicillin resistant Staphylococcus aureus by solid-state NMR. AB - Compositional analysis of the peptidoglycan (PG) of a wild-type methicillin resistant Staphylococcus aureus and its fem-deletion mutants has been performed on whole cells and cell walls using stable-isotope labeling and rotational-echo double-resonance NMR. The labels included [1-(13)C,(15)N]glycine and l-[epsilon (15)N]lysine (for a direct measure of the number of glycyl residues in the bridging segment), [1-(13)C]glycine and l-[epsilon-(15)N]lysine (concentration of bridge links), and d-[1-(13)C]alanine and [(15)N]glycine (concentrations of cross links and wall teichoic acids). The bridging segment length changed from 5.0 glycyl residues (wild-type strain) to 2.5 +/- 0.1 (FemB) with modest changes in cross-link and bridge-link concentrations. This accurate in situ measurement for the FemB mutant indicates a heterogeneous PG structure with 25% monoglycyl and 75% triglycyl bridges. When the bridging segment was reduced to a single glycyl residue 1.0 +/- 0.1 (FemA), the level of cross-linking decreased by more than 20%, resulting in a high concentration of open N-terminal glycyl segments. PMID- 19309107 TI - Transformation of a layered zinc phosphite to a three-dimensional open-framework structure with intersecting 16- and 12-ring channels. AB - A new layered zinc phosphite, H(2)tmdp x Zn(3)(HPO(3))(4) (tmdp = 4,4' trimethylenedipiperidine), was prepared in a flux of phosphorous acid without the addition of water as a solvent. The compound could be transformed into a new three-dimensional open-framework zinc phosphite, Zn(2)(H(2)O)(4)(HPO(3))(2) x H(2)O, with intersecting 16- and 12-ring channels at room temperature. PMID- 19309108 TI - Bis(zinc porphyrin) bridged by benzo orthocarbonates as a conformational switch under regulation of DABCO and a Cu+ ion. AB - A tweezer-type bis(zinc porphyrin) bridged by benzo orthocarbonates was synthesized and applied as a molecular conformational switch under regulation of DABCO and a Cu(+) ion. The switch property has been confirmed by (1)H NMR, UV-vis spectral titration, and HR-MS spectra method. PMID- 19309109 TI - Evidence for heme oxygenase activity in a heme peroxidase. AB - The heme peroxidase and heme oxygenase enzymes share a common heme prosthetic group but catalyze fundamentally different reactions, the first being H(2)O(2) dependent oxidation of substrate using an oxidized Compound I intermediate, and the second O(2)-dependent degradation of heme. It has been proposed that these enzymes utilize a common reaction intermediate, a ferric hydroperoxide species, that sits at a crossroads in the mechanism and beyond which there are two mutually exclusive mechanistic pathways. Here, we present evidence to support this proposal in a heme peroxidase. Hence, we describe kinetic data for a variant of ascorbate peroxidase (W41A) which reacts slowly with tert-butyl hydroperoxide and does not form the usual peroxidase Compound I intermediate; instead, structural data show that a product is formed in which the heme has been cleaved at the alpha-meso position, analogous to the heme oxygenase mechanism. We interpret this to mean that the Compound I (peroxidase) pathway is shut down, so that instead the reaction intermediate diverts through the alternative (heme oxygenase) route. A mechanism for formation of the product is proposed and discussed in the light of what is known about the heme oxygenase reaction mechanism. PMID- 19309110 TI - (15)N NMR relaxation data reveal significant chemical exchange broadening in the alpha-domain of human alpha-lactalbumin. AB - Human alpha-lactalbumin (alpha-LA), a 123-residue calcium-binding protein, has been studied using (15)N NMR relaxation methods in order to characterize backbone dynamics of the native state at the level of individual residues. Relaxation data were collected at three magnetic field strengths and analyzed using the model free formalism of Lipari and Szabo. The order parameters derived from this analysis are generally high, indicating a rigid backbone. A total of 46 residues required an exchange contribution to T(2); 43 of these residues are located in the alpha-domain of the protein. The largest exchange contributions are observed in the A-, B-, D-, and C-terminal 3(10)-helices of the alpha-domain; these residues have been shown previously to form a highly stable core in the alpha-LA molten globule. The observed exchange broadening, along with previous hydrogen/deuterium amide exchange data, suggests that this part of the alpha domain may undergo a local structural transition between the well-ordered native structure and a less-ordered molten-globule-like structure. PMID- 19309111 TI - Orbital hybridization and magnetic coupling of the A-site Cu spins in CaCu3B4O12 (B = Ti, Ge, and Sn) perovskites. AB - X-ray absorption spectroscopy (XAS) spectra near the O K-edge of A-site-ordered perovskite with A-site Cu(2+) (S = (1)/(2)) spins were measured. The spectra of ferromagnetic CaCu(3)Ge(4)O(12) and CaCu(3)Sn(4)O(12) showed hybridization between Cu 3d and O 2p orbitals, but magnetic circular dichroism measurement revealed that the O 2p orbital played a less important role in magnetic interaction. The XAS spectra of antiferromagnetic CaCu(3)Ti(4)O(12), on the other hand, showed strong hybridization of the Cu 3d, Ti 3d, and O 2p orbitals. These results demonstrated that direct exchange interaction of the Cu(2+) spins primarily determined the ferromagnetic ordering of CaCu(3)Ge(4)O(12) and CaCu(3)Sn(4)O(12), whereas the involvement of Ti 3d orbitals induced the antiferromagnetic property in CaCu(3)Ti(4)O(12). PMID- 19309112 TI - Morphology-dependent stimulated emission and field emission of ordered CdS nanostructure arrays. AB - Highly ordered CdS nanostructure arrays were successfully fabricated via a simple two-step metal-organic chemical vapor deposition process. Both stimulated emission and field-emission measurements were carried out in an attempt to understand the correlation between the morphologies, alignments, and emission performances of five ordered CdS nanostructure array types (well-aligned truncated nanocones, nanorods, cleft nanorods, quasi-aligned nanowires, and nanowires). The 1D CdS nanostructures of various types displayed notable differences in stimulated and field-emission performances. The stimulated emission strongly correlated with the structure alignment: the better the alignment, the lower the threshold. Both of the alignments and aspect ratios greatly affected the field-emission properties; the CdS emitters of higher aspect ratio and better alignment exhibited better field-emission performance. Thus the well-aligned CdS nanorod arrays had the lower threshold for stimulated emission, and quasi-aligned nanowire arrays produced the higher field-emission current and possessed the lower turn-on fields. PMID- 19309113 TI - Docking and MD simulations of the interaction of the tarantula peptide psalmotoxin-1 with ASIC1a channels using a homology model. AB - The interaction of the tarantula toxin PcTx1 with the hASIC1a ion channel is investigated here along homology modeling (using the crystal structure of the cASIC1 channel as a template and the known sequence of hASIC1a), automated docking (using the NMR solution structure of PcTx1), and molecular dynamics simulations (taking into account proton-binding sites), in what represents the first modeling and computational chemistry in the whole family of ASIC/DEG/ENaCs/FaNaCh channels. The results agree with binding and electrophysiological data for the interaction of mutant (125)I-PcTx1Y(N) with rASIC1a chimeras and PcTx1 itself with hASIC1a chimeras. They go even farther by revealing that only two hASIC1a subunits can be directly involved in the binding, to which four domains - instead of the only two identified by the experiments - participate. Mapping the closest lying amino acids of the homology model and PcTx1 can have heuristic value in stimulating ideas, software, and experimentation. PMID- 19309114 TI - Ligand prediction from protein sequence and small molecule information using support vector machines and fingerprint descriptors. AB - Support vector machine (SVM) database search strategies are presented that aim at the identification of small molecule ligands for targets for which no ligand information is currently available. In pharmaceutical research and chemical biology, this situation is faced, for example, when studying orphan targets or newly identified members of protein families. To investigate methods for de novo ligand identification in the absence of known three-dimensional target structures or active molecules, we have focused on combining sequence and ligand information for closely and distantly related proteins. To provide a basis for these investigations, a set of 11 protease targets from different families was assembled together with more than 2000 inhibitors directed against individual proteases. We have compared SVM approaches that combine protein sequence and ligand information in different ways and utilize 2D fingerprints as ligand descriptors. These methodologies were applied to search for inhibitors of individual proteases not taken into account during learning. A target sequence ligand kernel and, in particular, a linear combination of multiple target directed SVMs consistently identified inhibitors with high accuracy including test cases where homology-based similarity searching using data fusion and conventional SVM ranking nearly or completely failed. The SVM linear combination and target-ligand kernel methods described herein are intuitive and straightforward to adopt for ligand prediction against other targets. PMID- 19309115 TI - Beneficial effect of solubility enhancers on protein crystal nucleation and growth. AB - Crystallizing solutions of proteins often contain various nonelectrolyte additives that arise from the purification process of proteins or from the reagents employed in the screening kits. Currently, limited knowledge exists about the influence of these additives on the mechanisms underlying the crystallization process, in particular on the nucleation stage of crystals. To address this need, we studied crystallization of two proteins, D-xylose isomerase and chicken egg-white lysozyme, in small batches and in the presence of two solubility-enhancing additives, acetonitrile and glycerol. We have also measured the nucleation rates of crystals of these proteins in the presence and in the absence of acetonitrile using the method of initial rates. With the addition of the solubility enhancers, both proteins exhibited an increase in crystal nucleation at any given supersaturation. Solubility enhancing additives appear to lower the energy barrier to nucleation by influencing the strength of attraction between the protein molecules. We have characterized the quality of D-xylose isomerase crystals by determining the crystal mosaicity, which showed considerable improvement for crystals grown in the presence of additives. When compared to the crystals of chicken egg-white lysozyme, D-xylose isomerase crystals required higher supersaturations to nucleate. We attribute this result to the large size of the D-xylose isomerase molecule, which influences the energy barrier to nucleation by increasing the surface area of the critical nucleus. Contrary to the common expectation that reagents that solubilize the protein may hinder the crystallization process, our results suggest that solubility enhancers, in fact, can have a beneficial effect on the nucleation and growth of crystals. These findings are of importance in formulating successful strategies toward crystallizing new proteins. PMID- 19309116 TI - Modulation of morphology and conductivity of mixed-valence tetrathiafulvalene nanofibers by coexisting organic acid anions. AB - We describe here the facile preparation of conductive casting films consisting of mixed-valence tetrathiafulvalene (TTF) nanofibers. Self-assembled nanofibers of mixed-valence TTF with organic acid anions were grown during film deposition, and the conductivities of the resulting films were significantly altered by the type of the anion. In particular, the casting films containing heptafluorobutyrate, propanesulfonate, and undecanesulfonate work as semiconductors (10(-2) S/cm). It was revealed that anion species played a crucial role in the formation of the nanofibers and the morphology of the film was varied. PMID- 19309117 TI - On the possible role of surface elasticity in emulsion stability. AB - We have measured the short-time and long-time elastic responses to compression of various types of surfactant layers adsorbed at oil-water interfaces. We prepared reasonably monodisperse oil-in-water emulsions with the same surfactants and monitored the time evolution of the emulsion droplets' diameter. We used a broad variety of surfactants (cationic, nonionic, and small polymers) and alkanes with different chain lengths. The emulsion drop size evolution is first controlled by Ostwald ripening and later on by drop coalescence, the later step being quite short. The overall emulsion lifetime is therefore dominated by ripening and for a given oil appears well correlated with the low-frequency surface elasticity as expected (and not with the high-frequency one, which is expected to control coalescence). When the oil chain length is changed, the stability is related more to the oil solubility in water, which also controls ripening. The overall results demonstrate the great importance of surface elasticity in emulsion stability. PMID- 19309118 TI - Hierarchical micellar structures from amphiphilic invertible polyesters: 1H NMR spectroscopic study. AB - The environment-dependent behavior of invertible polyesters has been studied by 1H NMR spectroscopy. In dilute toluene solutions, the micelle exterior is made up of the lipophilic fragments, and the interior consists of the hydrophilic constituents. The polyester inverts the structure in an aqueous medium to form micelles with a hydrophobic inner part and a hydrophilic outer part. Increasing polyester concentration leads to the formation of hierarchical structures both in toluene and in an aqueous medium as a result of the aggregation of unimolecular micelles and the formation of hydrophilic and lipophilic domains. On the contrary, no unimolecular micelles or micellar aggregation has been observed in acetone or chloroform. PMID- 19309119 TI - Assessment of QM/MM scoring functions for molecular docking to HIV-1 protease. AB - We explore the ability of four quantum mechanical (QM)/molecular mechanical (MM) models to accurately identify the native pose of six HIV-1 protease inhibitors and compare them with the AMBER force field and ChemScore and GoldScore scoring functions. Three QM/MM scoring functions treated the ligand at the HF/6-31G*, AM1d, and PM3 levels; the fourth QM/MM function modeled the ligand and active site at the PM3-D level. For the discrimination of native from non-native poses, solvent-corrected HF/6-31G*:AMBER and AMBER functions exhibited the best overall performance. While the electrostatic component of the MM and QM/MM functions appears important for discriminating the native pose of the ligand, the polarization contribution in the QM/MM functions was relatively insensitive to a ligand's binding mode and, for one ligand, actually hindered discrimination. The inclusion of a desolvation penalty, here using a generalized Born solvent model, improved discrimination for the MM and QM/MM methods. There appeared to be no advantage to binding mode prediction by incorporating active site polarization at the PM3-D level. Finally, we found that choice of the protonation state of the aspartyl dyad in the HIV-1 protease active site influenced the ability of scoring methods to determine the native binding pose. PMID- 19309120 TI - Precise seed-mediated growth and size-controlled synthesis of palladium nanoparticles using a green chemistry approach. AB - In this paper, we present a "green" and size-controlled seed-mediated growth method by which differently sized palladium (Pd) nanoparticles, spanning from 3.4 to 7.6 nm, with an increment of 1.4 nm, were synthesized. Monodisperse Pd nanoparticles (ca. 3.4 nm, standard deviation = 0.7 nm) were first synthesized and stabilized in an aqueous solution at 95 degrees C using nontoxic ascorbic acid and sodium carboxymethyl cellulose (CMC) as reducing agent and capping agent, respectively. These Pd nanoparticles were subsequently employed as seeds on the surface of which fresh Pd (2+) ions were reduced by the weak reducing agent ascorbic acid. Optimal conditions were determined that favored the homogeneous and sequential accumulation of Pd atoms on the surface of the Pd seeds, rather than the formation of new nucleation sites in the bulk growth solution, thereby achieving atomic-level control over particle sizes. The adsorbed CMC molecules did not inhibit the addition of Pd atoms onto the seeds during the growth but provided stabilization of the Pd nanoparticles in aqueous solution after the growth. Potential mechanisms that underpin this seed-mediated growth process are provided and discussed. One advantage of this seed growth process is that it provides stoichiometric control over the size of the Pd nanoparticles by simply varying Pd(2+) added during the growth stage. Furthermore, the use of ecologically friendly reagents, such as water (solvent), CMC (capping agent), and ascorbic acid (reducing agent), in both the seed synthesis and subsequent seed-mediated growth provides both "green" and economic attributes to this process. PMID- 19309121 TI - Rearrangemements in piperidine-derived nitrogen-centered radicals. A quantum chemical study. AB - Hydrogen migration reactions in piperidine radicals and their protonated counterparts were studied by quantum chemical calculations. G3B3 and G3(MP2)-RAD levels of theory were used as reference procedures in order to evaluate the efficiency of other computational models. In the gas phase, the 1,4-[N<-->C]-H and 1,4-[C<-->C]-H shifts are the most feasible rearrangements in the piperidine radical cation and neutral piperidine radical, respectively. However, if one explicit water molecule is well placed to facilitate hydrogen migrations, the 1,2 [N<-->C]-H shift becomes the most favorable process in both cases. Three different water-catalyzed [N<-->C]-H shift mechanisms were considered for piperidine radical cation, and only one is found to be operative in the case of neutral piperidine radical. We found that explicit solvation and protonation of piperidine-derived radicals strongly influence the overall mechanism of hydrogen migration reactions. PMID- 19309122 TI - Cobalt-catalyzed cross-coupling between in situ prepared arylzinc halides and 2 chloropyrimidine or 2-chloropyrazine. AB - A cobalt-catalyzed cross-coupling of aryl halides with 2-chloropyrimidines or 2 chloropyrazines is reported in satisfactory to high yields. The key step of this procedure is the formation of aromatic organozinc reagents and their coupling with 2-chlorodiazines using the same cobalt halide as catalyst and Zn dust under mild reaction conditions. This new cobalt-catalyzed coupling reaction represents a practical and interesting alternative to previously known methods for the synthesis of 2-aryldiazines. PMID- 19309123 TI - New fluorinated peptidomimetics through tandem aza-michael addition to alpha trifluoromethyl acrylamide acceptors: synthesis and conformational study in solid state and solution. AB - A range of partially modified retro (PMR) psi[NHCH(2)] peptide mimetics containing a hydrolytically stable CH(2)CH(CF(3))CO unit have been synthesized. The first kind of peptidomimetics is obtained from the highly efficient aza Michael addition of different amines to alpha-trifluoromethyl acrylamide acceptors. Subsequent deprotection of the amino group furnishes the key common intermediate for the synthesis of other families of peptidomimetics: dipeptides, tripeptides, peptidomimetics containing a urea moiety, and structures containing two units of alpha-trifluoromethyl-beta(2)-alanine. Finally, a conformational study of several of the newly synthesized peptidomimetics, performed with the aid of X-ray analysis and NMR techniques, shows a beta-turn-like conformation for the structures both in the solid state and in solution. PMID- 19309124 TI - Optimized synthesis and enhanced efficacy of novel triplex-forming camptothecin derivatives based on gimatecan. AB - Sequence-specific camptothecins are useful tools to inhibit specifically gene expression. The camptothecins are attached to the 3' end of triplex-forming oligonucleotides (TFO), sequence-specific DNA ligands that position the camptothecin moiety exclusively in proximity to their binding site. We studied here different gimatecan derivatives or analogues, a potent lipophilic camptothecin compound in clinical trials. We optimized the synthesis procedure in order to increase the yields and the purity and obtain the conjugates on a large scale. The greatly improved synthesis is now based on the conjugation of a bromoalkyl analogue of gimatecan to the 3' phosphorothioate of the TFO. We showed that the most efficient conjugate, both in vitro and in HeLa cells, bears the TFO on position 7 of the gimatecan analogue, and it is more efficient than the previous camptothecin conjugates. In addition, the gimatecan-like moiety at the 3' end of the TFO protects from nuclease degradation. PMID- 19309125 TI - Evaluation of cation influence on the formation and extraction capability of ionic-liquid-based aqueous biphasic systems. AB - In addition to the large range of applications proposed in literature, ionic liquids (ILs) have been recently reported to be able to form aqueous biphasic systems (ABS). They could thus be interesting media in biotechnological applications for the separation and purification of vital biomolecules. Therefore, in this work, a systematic study involving a large number of imidazolium-based ILs was conducted to provide new information related to ILs' ABS-promoting capability and extraction ability. For that purpose, the influence of the number of alkyl groups present at the cation, the cation side alkyl chain length, and the presence of double bonds, aromatic rings, and hydroxyl groups on this alkyl chain were evaluated. Ternary phase diagrams of the ABS formed by these ILs and K(3)PO(4) and the respective tie-lines were measured and presented. The ABS here investigated were further characterized for the first time accordingly to their extractive potential for amino acids, where l-tryptophan was selected as a model biomolecule. The partition coefficients here obtained were shown to be substantially larger than those observed in conventional ABS, demonstrating therefore the fine potential of IL-based ABS for biomolecules separation and purification. PMID- 19309126 TI - Chiral sum frequency spectroscopy of thin films of porphyrin J-aggregates. AB - Thin films of chiral porphyrin J-aggregates have been studied by vibrationally and electronically doubly resonant sum frequency generation (SFG) spectroscopy. It was revealed that the chiral supramolecular structures of porphyrin aggregates in solutions were retained in the thin film samples, and their chirality was determined by using chiral vibrational SFG spectroscopy. Electronic resonance profiles of some vibrational bands in achiral and chiral SFG were different from each other, and both were distinct from electronic absorption spectra. To account for these peculiar profiles, we have proposed interference effects of Raman tensor components in achiral and chiral SFG susceptibilities, which is analogous to that of resonance Raman scattering. PMID- 19309127 TI - Measurement of DNA morphological parameters at highly entangled regime on surfaces. AB - The morphology of circular DNA deposited from a solution on the mica surface is analyzed from the power spectrum density (PSD) of the atomic force microscopy (AFM) images. Sample morphology is modulated in a broad range of concentration C from isolated molecules to highly entangled networks. DNA exhibits a multiaffine behavior with two correlation length scales: the persistence length P which remains constant ( approximately 50 nm) within the C range and the intermolecular distance xi which exhibits a decay with increasing C. Applying a diffusion based model in which xi scales as xi approximately D(-0.25).C(-0.5), we extracted the DNA diffusion coefficient D approximately 2 x 10(-7) cm(2)/s. This value is consistent with a high-molecular-weight plasmid DNA supercoiled in the solution. PMID- 19309128 TI - Role of charge transfer in the structure and dynamics of the hydrated proton. AB - Although it has long been recognized that multiple water molecules strongly associate with an extra proton in bulk water, some models and conceptual frameworks continue to utilize the classical hydronium ion (H(3)O(+)) as a fundamental building block. In this work, the nature of the hydronium ion in aqueous systems is examined using an ab initio energy decomposition analysis (EDA) that evaluates both the magnitude of and energetic stabilization due to charge transfer among H(3)O(+) and the surrounding water molecules. The EDA is performed on structures extracted from dynamical bulk-phase simulations and used to determine how frequently the pure hydronium ion, where the excess charge is primarily localized on H(3)O(+), occurs under dynamic conditions. The answer is essentially never. The energetic stabilization of H(3)O(+) due to charge delocalization to neighboring water molecules is found to be much larger (16-49 kcal/mol) than for other ions (even Li(+)) and to constitute a substantial portion (20-52%) of the complex's binding energy. The charge defect is also shown to have intrinsic dynamical asymmetry and to display dynamical signatures that can be related to features appearing in IR spectra. PMID- 19309129 TI - Mechanism of base-promoted dehydrochlorination of pentachloroethane: concerted or stepwise? AB - This paper reports a first-principles molecular dynamics simulation study of the microscopic mechanism of the base-catalyzed dehydrochlorination of pentachloroethane. So far the nature of the mechanism of this reaction is not understood: the concerted and stepwise mechanisms are under debate. By combining ab initio molecular dynamics with the method of constraints, we determine the reaction mechanism and associated free energy profile. We find that the reaction barrier is in good agreement with experimental findings and reveal that the reaction proceeds via a concerted mechanism. Our simulations provide no evidence for the presence of carbanion intermediate indicative of the stepwise pathway. This microscopic understanding will provide new implications for understanding the reduction of polyhalogenated alkanes and rational design of effective materials to treat these contaminants. PMID- 19309130 TI - Phase sensitive control of vibronic guest-host interaction: Br2 in Ar matrix. AB - Vibronic progressions are programmed into a pulse shaper which converts them via the inherent Fourier transformation into a train of femtosecond pulses in time domain for chromophore excitation. Double pulse results agree with phase sensitive wave packet superposition from a Michelson interferometer which delivers coherence times with high reliability. Spectral resolution of 1 nm and a spacing of around 4 nm within the 20 nm envelope centered at 590 nm delivers a train of seven phase-controlled 40 fs subpulses separated by 250 fs. Combs adjusted to the zero phonon lines (ZPL) and phonon sidebands (PSB) of the B state vibronic progression are reproduced in the chromophore for a coherent subpulse accumulation. B state ZPL wave packet dynamics dominates in pump-probe spectra due to its coherence despite an overwhelming but incoherent A state contribution in absorption. PSB comb accumulation is also phase sensitive and demonstrates coherence within several 100 matrix degrees of freedom in the vicinity. PMID- 19309131 TI - Another treatment of fluorescence polarization microspectroscopy and imaging. AB - We here discuss a general (symmetry adapted) treatment for one-photon-excitation time-resolved fluorescence polarization microspectroscopy (TRFPM) at combined wide-angular excitation and detection apertures that correctly couples the principles of the optics of objective lenses with the principles of fluorescence spectroscopy with polarized light. The treatment is unified in the sense that it covers the electromagnetic description of focusing a linearly polarized beam of exciting light (diffraction theory, DT) and the description of the same problem in terms of the meridional plane properties (MPP) of the objective lenses (geometrical optics). It is shown that both approaches are quantitatively equivalent from the point of view of the polarization effects in typical TRFPM experiments on linear absorbers, despite the fact that in the MPP treatment the region of focus is treated as a pointlike object, while in the DT method the region of focus is characterized by a three-dimensional (3D) inhomogeneous electromagnetic field distribution, of generally ellipsoidal polarization at different points of the focus. This finding is essentially important from the point of view of the experimental practice because the MPP treatment is based on two very simple trigonometric expressions, in evident contrast to the DT method, in which the high-aperture focusing is described in terms of three complicated 3D integrals involving the Bessel functions of the first kind. A few words of comment are added on a similar problem in the case of nonlinear one-photon absorbers (e.g., chiral fluorophores). We discuss the synthetic fluorescence decays for the wide-field- and evanescent-wave-excitation confocal (or wide field) detection fluorescence polarization microspectroscopy and imaging, which indicate the right experimental protocols for the kinetic and dynamic fluorescence polarization microspectroscopic studies. The manifestations of the effects resulting from the application of the wide-angular excitation and/or detection apertures are displayed and discussed in a systematic way. A few words of comment are added on the application of the symmetry adapted calibration (SAC) method to TRFPM experiments. A very important aim of this article is to provide a correct and more complete description of fluorescence polarization microspectroscopy and imaging of macroscopically isotropic media (i.e., solutions, solutions of labeled macromolecules, membrane suspensions, or biological cells), that can be immediately applied in the experimental practice in the life and medical sciences and also in different areas of nano(bio)technology. PMID- 19309132 TI - Identification of serine/threonine kinase substrates in the human pathogen group B streptococcus. AB - All living organisms respond to changes in their internal and external environment for their survival and existence. Signaling is primarily achieved through reversible phosphorylation of proteins in both prokaryotes and eukaryotes. A change in the phosphorylation state of a protein alters its function to enable the control of cellular responses. A number of serine/threonine kinases regulate the cellular responses of eukaryotes. Although common in eukaryotes, serine/threonine kinases have only recently been identified in prokaryotes. We have described that the human pathogen Group B Streptococcus (GBS, Streptococcus agalactiae) encodes a single membrane-associated, serine/threonine kinase (Stk1) that is important for virulence of this bacterium. In this study, we used a combination of phosphopeptide enrichment and mass spectrometry to enrich and identify serine (S) and threonine (T) phosphopeptides of GBS. A comparison of S/T phosphopeptides identified from the Stk1 expressing strains to the isogenic stk1 mutant indicates that 10 proteins are potential substrates of the GBS Stk1 enzyme. Some of these proteins are phosphorylated by Stk1 in vitro and a site-directed substitution of the phosphorylated threonine to an alanine abolished phosphorylation of an Stk1 substrate. Collectively, these studies provide a novel approach to identify serine/threonine kinase substrates for insight into their signaling in human pathogens like GBS. PMID- 19309133 TI - Intramolecular addition of cysteine thiyl radical to phenylalanine and tyrosine in model peptides, Phe (CysS*) and Tyr(CysS*): a computational study. AB - Density Functional Theory (DFT) calculations were carried out to evaluate the potential for intramolecular addition of peptide cysteine (Cys) thiyl radicals (CysS(*)) to aromatic amino acids (Phe and Tyr) in water. These calculations yielded cyclic conformations, in which pi-complexes were more stable than cyclohexadienyl radicals in water. In these pi-complexes, the C(2)-S distances were significantly shorter compared to the C(1)-S and C(3)-S distances. Comparable results on the relative stabilities were obtained for model calculations for the addition of HS(*)/CH(3)S(*) to toluene and p-hydroxytoluene. The adduct of thiyl radicals with Phe was more stable than that with Tyr, and the stabilization energies depended on the C-terminal substituents. PMID- 19309135 TI - Loosening and reorganization of fluid phospholipid bilayers by chloroform. AB - The mixing behavior of an exchangeable phospholipid (A) with an exchangeable sterol (B) in host bilayers made from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) containing varying concentrations of cholesterol has been examined via the nearest-neighbor recognition method. At low sterol concentrations (i.e., 2.5 mol %), the mixing between A and B is close to ideal. Incremental increases in the sterol concentration to 40 mol % led to net increases in the affinity between A and B. Similar mixing experiments that were carried out in the presence of chloroform showed a leveling effect, where moderate sterol-phospholipid affinity was observed in all cases. These results, together with the fact that the number of chloroform molecules that are absorbed per phospholipid is essentially constant and independent of the sterol content, support a model in which chloroform favors solvation of the phospholipids and a common membrane state is produced. Fluorescence measurements and Raman spectra have also shown that chloroform significantly loosens both cholesterol-poor and cholesterol-rich membranes made from DPPC. In a broader context, these results suggest a fundamentally new mechanism of anesthesia, where the anesthetic, by solvating the lipid components, profoundly changes the lateral organization of the lipid framework. PMID- 19309136 TI - Interpretation of Raman and Raman optical activity spectra of a flexible sugar derivative, the gluconic acid anion. AB - Raman scattering and its polarized extension, Raman optical activity (ROA), are commonly used for monitoring of molecular conformational equilibria in solutions. This is complicated for saccharides due to extensive motions of the hydroxyl groups and other molecular parts. Standard interpretation procedures involving ab initio spectral simulations for a limited set of conformers are not adequate. In this study, a more general approach is proposed for the gluconic acid anion taken as a model compound, where quantum simulations of the spectra are directly coupled with molecular dynamics (MD) techniques. Such a multiscale approach reveals how the structural information is encoded in the broadened spectral lines. The spectra were simulated for solvent-solute clusters generated by MD. Conformational averaging was enabled by a limited library of conformers for which the spectral parameters could be calculated ab initio and moved on the MD geometries by Cartesian coordinate tensor transfer techniques. The B3LYP/CPCM/6 31+G** approximation was used as a default for computation of the source force fields and polarizability derivatives. The spectra thus obtained relatively faithfully reproduced most of the experimental features. The Amber and polarizable Amoeba MD force fields produced similar results; application of the latter, however, was limited by the long time necessary to achieve a converged conformational equilibrium. Both MD simulation and spectral averaging suggest that the hydroxyl groups as well as the backbone C-C bonds rotate relatively freely, with some restrictions in the vicinity of the carboxyl group. In spite of the averaging, spectral response of characteristic vibrational normal mode families, such as CH and OH bending, can clearly be identified in the spectra. The simulations thus confirm the experimental fact that flexible saccharides exhibit significant vibrational activity that reveals precious information about molecular structure and dynamics encoded in the Raman and ROA spectral shapes. PMID- 19309138 TI - Joint experimental and theoretical studies of the mechanism for the gas phase elimination kinetics of methyl 2,2-dimethyl-3-hydroxypropionate. AB - Methyl 2,2-dimethyl-3-hydroxypropionate was found to decompose, in a static system, mainly to methyl isobutyrate and formaldehyde. The reaction rates were affected in packed and unpacked clean Pyrex vessels, demonstrating little but significant surface effect. However, in vessels seasoned with allyl bromide this reaction was homogeneous and unimolecular and followed a first-order law. The working temperature range was 349-410 degrees C and the pressure range was 64-162 Torr. The variation of the rate coefficient with temperature is expressed by the following Arrhenius expression: log k(1) (s(-1)) = [(11.43 +/- 0.57) - (180.4 +/- 7.2) kJ mol(-1)] x (2.303RT)(-1). Methyl 2,2-dimethyl-3-hydroxypropionate was found to be 1.4 times greater in the rate of elimination than methyl 3 hydroxypropionate. Apparently, steric acceleration may be considered responsible in the process of decomposition. The theoretical calculation of the kinetics and thermodynamics parameters, at the B3LYP/6-211G** level of theory, are in reasonably good agreement with the experimental values obtained. These calculations imply a molecular mechanism involving a concerted nonsynchronous transition state where abstraction of the hydroxyl hydrogen by the oxygen of the carbonyl ester is a determining factor and the transition state is late in the reaction coordinate. PMID- 19309137 TI - Lipophilic bisphosphonates as dual farnesyl/geranylgeranyl diphosphate synthase inhibitors: an X-ray and NMR investigation. AB - Considerable effort has focused on the development of selective protein farnesyl transferase (FTase) and protein geranylgeranyl transferase (GGTase) inhibitors as cancer chemotherapeutics. Here, we report a new strategy for anticancer therapeutic agents involving inhibition of farnesyl diphosphate synthase (FPPS) and geranylgeranyl diphosphate synthase (GGPPS), the two enzymes upstream of FTase and GGTase, by lipophilic bisphosphonates. Due to dual site targeting and decreased polarity, the compounds have activities far greater than do current bisphosphonate drugs in inhibiting tumor cell growth and invasiveness, both in vitro and in vivo. We explore how these compounds inhibit cell growth and how cell activity can be predicted based on enzyme inhibition data, and using X-ray diffraction, solid state NMR, and isothermal titration calorimetry, we show how these compounds bind to FPPS and/or GGPPS. PMID- 19309139 TI - Shear-driven motion of supported lipid bilayers in microfluidic channels. AB - In this work, we demonstrate how a lateral motion of a supported lipid bilayer (SLB) and its constituents can be created without relying on self-spreading forces. The force driving the SLB is instead a viscous shear force arising from a pressure-driven bulk flow acting on the SLB that is formed on a glass wall inside a microfluidic channel. In contrast to self-spreading bilayers, this method allows for accurate control of the bilayer motion by altering the bulk flow in the channel. Experiments showed that an egg yolk phosphatidylcholine SLB formed on a glass support moved in a rolling motion under these shear forces, with the lipids in the upper leaflet of the bilayer moving at twice the velocity of the bilayer front. The drift velocity of different lipid probes in the SLB was observed to be sensitive to the interactions between the lipid probe and the surrounding molecules, resulting in drift velocities that varied by up to 1 order of magnitude for the different lipid probes in our experiments. Since the method provides a so far unattainable control of the motion of all molecules in an SLB, we foresee great potential for this technique, alone or in combination with other methods, for studies of lipid bilayers and different membrane-associated molecules. PMID- 19309140 TI - Thymine dimerization in DNA model systems: cyclobutane photolesion is predominantly formed via the singlet channel. AB - UV-induced formation of cylcobutane pyrimidine dimers (CPD) in all thymine DNA models have been studied by femtosecond IR spectroscopy. CPDs are shown to form within approximately 1 ps during the decay of the initially excited (1)pi pi * state. The quantum yields phi(D)(ps) determined after the (1)pi pi * decay equal the final yield phi(D)(cw). This gives evidence for a predominance of the singlet channel in CPD formation. PMID- 19309141 TI - Supramolecular cocaine-aptamer complexes activate biocatalytic cascades. AB - The anti-cocaine aptamer was fragmented into two nucleic acids, (1) and (2). The nucleic acid (1) was tethered at its 5'-end to aminoethyl nicotinamide adenine dinucleotide, amino-NAD(+), or to horseradish peroxidase, HRP. The nucleic acid (2) was functionalized at its 3'-end with alcohol dehydrogenase, AlcDH, or with glucose oxidase, GOx. In the presence of cocaine, the supramolecular NAD(+)/AlcDH/cocaine-aptamer complex is formed, and the biocatalytic oxidation of ethanol is activated. Similarly, in the presence of cocaine, the GOx/HRP/cocaine aptamer complex is formed, and this activates the biocatalytic cascade where glucose is oxidized by GOx to yield gluconic acid and H(2)O(2), and the resulting hydrogen peroxide activates the HRP-biocatalyzed oxidation of 2,2'-azinobis-(3 ethylbenzthiazoline-6-sulfonate), ABTS(2-). The systems may be considered as biomimetic prototypes for systems biology. PMID- 19309143 TI - Conformational study of the PCU cage monopeptide: a key role of some force-field parameters. AB - As had been previously reported standard AMBER parameters do not reproduce adequately ab initio calculations performed on the pentacycloundecane cage residue. The analysis of the geometries of the minimum energy structures compared with other residues allowed us to suspect the importance of bond stretching parameters in this case. Accordingly, we proceeded to compute new stretching and bending parameters from density functional theory calculations. Accordingly, the reliability of these parameters have been validated by predicting the correct conformational Ramachandran map. This is an example where the geometry of the side chain constrains the geometry of the residue backbone in such a way that you can get the right conformational profile by simple modification of some stretching and bending parameters. PMID- 19309142 TI - Determinants of catalytic power and ligand binding in glutamate racemase. AB - Glutamate racemases (EC 5.1.1.3) catalyze the cofactor-independent stereoinversion of D- and L-glutamate and are important for viability in several gram-negative and -positive bacteria. As the only enzyme involved in the stereoinversion of L- to D-glutamate for peptidoglycan biosynthesis, glutamate racemase is an attractive target for the design of antibacterial agents. However, the development of competitive tight-binding inhibitors has been problematic and highly species specific. Despite a number of recent crystal structures of cofactor-independent epimerases and racemases, cocrystallized with substrates or substrate analogues, the source of these enzymes' catalytic power and their ability to acidify the C alpha of amino acids remains unknown. The present integrated computational and experimental study focuses on the glutamate racemase from Bacillus subtilis (RacE). A particular focus is placed on the interaction of the glutamate carbanion intermediate with RacE. Results suggest that the reactive form of the RacE-glutamate carbanion complex, vis-a-vis proton abstraction from C alpha, is significantly different than the RacE-D-glutamate complex on the basis of the crystal structure and possesses dramatically stronger enzyme-ligand interaction energy. In silico and experimental site-directed mutagenesis indicates that the strength of the RacE-glutamate carbanion interaction energy is highly distributed among numerous electrostatic interactions in the active site, rather than being dominated by strong hydrogen bonds. Results from this study are important for laying the groundwork for discovery and design of high-affinity ligands to this class of cofactor-independent racemases. PMID- 19309144 TI - Oxidation of aluminum particles in the presence of water. AB - Oxidation of spherical aluminum powder was investigated in mixed argon-oxygen steam atmospheres by thermogravimetric measurements at heating rates between 1 and 20 K/min and up to 1100 degrees C. The observed oxidation behavior in the presence of steam differs markedly from oxidation in dry oxygen. Oxidation in steam is complete near 1000 degrees C vs 1500 degrees C in dry oxygen. Furthermore, in steam, a stepwise weight change is observed at the melting point of aluminum, while no such step can be distinguished in dry oxygen. The complete oxidation observed at a lower temperature in steam as compared to dry oxygen is explained by the stabilization of the gamma polymorph of the surface oxide in the presence of water so that a denser and slower growing alpha-alumina does not form until higher temperatures. Experiments in mixed oxygen/steam oxidizers showed that the size of the oxidation step observed upon aluminum melting only correlates with the concentration of steam in the atmosphere. This may be interpreted as the effect of transient porosity, the degree of which is controlled by the steam concentration, or the surface oxide stressed by the expanding melting metal core may behave as a semipermeable membrane where hydrous species have significantly higher diffusion rates than oxygen. A clear distinction cannot be drawn, and further research is warranted. Preliminary results on isoconversion processing of the oxidation kinetics are presented. PMID- 19309145 TI - Self-assembly of dendronized triphenylenes into helical pyramidal columns and chiral spheres. AB - The synthesis and structural and retrostructural analyses of a library containing 10 triphenylenes functionalized with self-assembling benzyl ether and phenyl propyl ether dendrons are reported. These dendronized triphenylenes adopt a crown rather than discotic conformation. Their crown conformation mediates the self assembly of the discotic triphenylene unit in helical pyramidal columns and in chiral spheres. The chiral spheres are generated from short segments of helical pyramidal columns that are spherically distorted. Therefore, the chirality of the sphere is determined by a short helical pyramidal column that represents the inner part of the supramolecular sphere. Both the helical pyramidal columns and the chiral spheres represent supramolecular architectures that were self assembled for the first time from discotic molecules. The helical pyramidal columns self-organize in various hexagonal and rectangular lattices, while the chiral spheres self-organize into cubic and tetragonal periodic arrays and into a quasiperiodic 12-fold liquid quasicrystal. The helical sense of the helical pyramidal columns and of helical spheres is selected by a stereocenter that can be incorporated either in the alkyl groups of the dendron or in the triphenylene part of the dendritic crown via donor-acceptor interactions. The self-assembly process of the dendronized triphenylene donor can be programmed by a new supramolecular "polymer effect" generated by donor-acceptor interactions. PMID- 19309146 TI - Total synthesis of N-acetylglucosamine-1,6-anhydro-N-acetylmuramylpentapeptide and evaluation of its turnover by AmpD from Escherichia coli. AB - The bacterial cell wall is recycled extensively during the course of cell growth. The first recycling event involves the catalytic action of the lytic transglycosylase enzymes, which produce an uncommon 1,6-anhydropyranose moiety during separation of the muramyl residues from the peptidoglycan, the major constituent of the cell wall. This product, an N-acetyl-beta-D-glucosamine-(1- >4)-1,6-anhydro-N-acetyl-beta-D-muramylpeptide, is either internalized to initiate the recycling process or diffuses into the milieu to cause stimulation of the pro-inflammatory responses by the host. We report the total syntheses of N acetyl-beta-D-glucosamine-(1-->4)-1,6-anhydro-N-acetyl-beta-D-muramyl-L-Ala-gamma D-Glu-meso-DAP-D-Ala-D-Ala (compound 1, the product of lytic transglycosylase action on the cell wall of gram-negative bacteria) and N-acetyl-beta-D glucosamine-(1-->4)-1,6-anhydro-N-acetyl-beta-D-muramyl-L-Ala-gamma-D-Glu-L-Lys-D Ala-D-Ala (compound 2, from lytic transglycosylase action on the cell wall of gram-positive bacteria). The syntheses were accomplished in 15 linear steps. Compound 1 is shown to be a substrate of the AmpD enzyme of the gram-negative bacterium Escherichia coli, an enzyme that removes the peptide from the disaccharide scaffold in the early cytoplasmic phase of cell wall turnover. PMID- 19309147 TI - Evaluation and comparison of the antioxidative potency of various carbohydrates using different methods. AB - A detailed analysis of the antioxidative activity of 12 carbohydrates including chondroitin sulfate, fucoidan, agaro-oligosaccharide, 2-deoxy-scyllo-inosose (DOI), Galbeta1-4DOI, D-glucuronic acid, chitobiose, D-mannosamine, D galactosamine, D-glucosamine, heparin, and colominic acid was performed using four established methods: 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, ferric reducing antioxidant power (FRAP) assay, superoxide dismutase (SOD) activity assay, and the deoxyribose method. Ascorbic acid and/or catechin were used as positive standards. In the DPPH radical scavenging activity measurements, fucoidan, DOI, and Galbeta1-4DOI showed remarkable levels of activity, although at lower levels than ascorbic acid. The SOD assay revealed that DOI, Galbeta1 4DOI, and agaro-oligosaccharide had high antioxidant activity, with DOI and Galbeta1-4DOI notably showing almost half of the antioxidative potency of ascorbic acid. Using the deoxyribose method, chitobiose and heparin showed the highest hydroxyl radical scavenging activity, followed by chondroitin sulfate, colominic acid, Galbeta1-4DOI, and d-glucosamine. Given that 11 of the carbohydrates analyzed share a common structure, agaro-oligosaccharide being the exception, we propose from our current results that at least one amino, carboxyl, carbonyl, or sulfonyl group is required, but is not in itself sufficient, for carbohydrates to function as antioxidants. PMID- 19309148 TI - Interactive effects of sulfur and nitrogen supply on the concentration of sinigrin and allyl isothiocyanate in Indian mustard (Brassica juncea L.). AB - Food derived from Brassica species is rich in glucosinolates. Hydrolysis of these compounds by myrosinase yields isothiocyanates and other breakdown products, which due to their pungency represent the primary purpose of Indian mustard cultivation. Strong interactive effects of S (0.0, 0.2, and 0.6 g pot(-1)) and N (1, 2, and 4 g pot(-1)) supply on growth, seed yield, and the concentrations of glucosinolates and isothiocyanates in seeds were observed in growth experiments, reflecting the involvement of S-containing amino acids in both protein and glucosinolate synthesis. At intermediate S supply, a strong N-induced S limitation was apparent, resulting in high concentrations of sinigrin (12 micromol g(-1) of DM) and allyl isothiocyanate (213 micromol kg(-1) of DM) at low N supply only. Myrosinase activity in seeds increased under low N and low S supply, but the results do not suggest that sinigrin functions as a transient reservoir for S. PMID- 19309149 TI - Development of a gas diffusion multicommuted flow injection system for the determination of sulfur dioxide in wines, comparing malachite green and pararosaniline chemistries. AB - A flow system based on the multicommutation concept was developed for the determination of free and total sulfur dioxide in table wines, exploiting gas diffusion separation and spectrophotometric detection. The system allowed the comparison of malachite green and pararosaniline chemistries, using the same manifold configuration. Free and total SO(2) were determined within the ranges 1.00-40.0 and 25.0-250 mg L(-1), at determination throughputs of 25 and 23 h(-1), respectively. Employing the malachite green reaction, detection limits of 0.3 and 0.8 mg L(-1) were attained for free and total SO(2), respectively. Pararosaniline chemistry provided detection limits of 0.6 mg L(-1) for free SO(2) and 0.8 mg L( 1) for total SO(2). Relative standard deviations better than 1.8 and 1.4% were obtained by the malachite green and pararosaniline reactions, respectively. With regard to the two tested chemistries, 18 wines were analyzed and the results achieved by the pararosaniline reaction compared better with those furnished by the recommended procedure. PMID- 19309150 TI - Evolution of volatile compounds during the development of cabernet sauvignon grapes (Vitis vinifera L.). AB - The evolution of volatile compounds was explored in grape berries at fortnightly intervals from fruit-set to late ripening to identify when biosynthetic pathways may be targeted for enhancement of grape and wine aroma. Stepwise linear discriminant analysis (SLDA) fully recognized patterns in berry physiological developmental stages with most of the variance (>99.0%) explained. The preveraison berry developmental stage was identified as a transition stage for volatile compound biosynthesis when most compounds were potentially sequestered to nonvolatile conjugates and berries lost their potential to synthesize esters and terpenes. Terpenes (predominantly eucalyptol, beta-caryophyllene, and alpha humulene) characterized early berry development, whereas benzene derivatives (2 phenylethanol and 2-phenylethanal) appeared toward late ripening. Furthermore, C(6) volatile compounds changed from acetate esters to aldehydes and finally to alcohols during early, middle, and late berry developmental stages, respectively. The dominance of alcohols in the late stages of berry development, preceded by aldehydes, offers an opportunity for alcohols to aldehydes ratios to be used in the prediction of harvest timing for enhanced grape and wine aroma. The evolution of volatile compounds during berry development suggests a greater dependency on enzyme activity and specificity than extent of fatty acid unsaturation. The dependence of the stage of berry development on the accumulation of the products of alcohol dehydrogenase (ADH), alcohol acetyl transferase (AAT), and enal isomerase enzyme activity from the lipoxygenase pathway raises possibilities for the manipulation of aroma profiles in grapes and wines. PMID- 19309151 TI - Comment on electrodeposited silicate films: importance of supporting electrolyte. AB - We have observed similar surface color changes of glassy carbon (GC) after anodic "electrodeposition" with the same electrolytes in the presence or the absence of silanes, indicating that sample color changes can arise from more than one thing and thus should not be used as the sole indicator of silica film formation. Nevertheless, the formation of silicate films during anodic electrodeposition is verified by X-ray photoelectron spectroscopy (XPS) measurements. PMID- 19309152 TI - Discovery of novel tricyclic full agonists for the G-protein-coupled niacin receptor 109A with minimized flushing in rats. AB - Tricyclic analogues were rationally designed as the high affinity niacin receptor G-protein-coupled receptor 109A (GPR109A) agonists by overlapping three lead structures. Various tricyclic anthranilide and cycloalkene carboxylic acid full agonists were discovered with excellent in vitro activity. Compound 2g displayed a good therapeutic index regarding free fatty acids (FFA) reduction and vasodilation effects in rats, with very weak cytochrome P450 2C8 (CYP2C8) and cytochrome P450 2C9 (CYP2C9) inhibition, and a good mouse pharmacokinetics (PK) profile. PMID- 19309153 TI - Evaluation of antioxidant properties of monoaromatic derivatives of pulvinic acids. AB - The natural mushroom pigment Norbadione A and three other pulvinic acids were shown by our group to display very efficient antioxidant properties by comparison with a collection of potent molecules including catechols, flavonoids, stilbenes, or coumarins. Despite numerous publications on robust and straightforward synthetic access to pulvinic acids by us and others, no report has been made to unravel the structure-activity relationships that govern the striking antioxidant activity. Herein is presented the synthesis of 18 diverse pulvinic acid derivatives and the study of their radical scavenging capacities by four different assays. The influence of each of the two phenyl rings, of their substituents and of the lateral chain on the antioxidant properties, was explored to reveal a simplified structure of excellent activity. These results, along with the absence of cytotoxicity, make the synthesized compounds interesting to evaluate for several biological activities and especially for anti-inflammatory effects and skin protection against UV induced oxidative stress. PMID- 19309154 TI - Development of novel aminoglycoside (NB54) with reduced toxicity and enhanced suppression of disease-causing premature stop mutations. AB - Nonsense mutations promote premature translational termination and represent the underlying cause of a large number of human genetic diseases. The aminoglycoside antibiotic gentamicin has the ability to allow the mammalian ribosome to read past a false-stop signal and generate full-length functional proteins. However, severe toxic side effects along with the reduced suppression efficiency at subtoxic doses limit the use of gentamicin for suppression therapy. We describe here the first systematic development of the novel aminoglycoside 2 (NB54) exhibiting superior in vitro readthrough efficiency to that of gentamicin in seven different DNA fragments derived from mutant genes carrying nonsense mutations representing the genetic diseases Usher syndrome, cystic fibrosis, Duchenne muscular dystrophy, and Hurler syndrome. Comparative acute lethal toxicity in mice, cell toxicity, and the assessment of hair cell toxicity in cochlear explants further indicated that 2 exhibits far lower toxicity than that of gentamicin. PMID- 19309155 TI - Synthesis and anti-inflammatory properties of 1alpha,25-dihydroxy-16-ene-20 cyclopropyl-24-oxo-vitamin D3, a hypocalcemic, stable metabolite of 1alpha,25 dihydroxy-16-ene-20-cyclopropyl-vitamin D3. AB - 1alpha,25(OH)(2)-16-ene-20-cyclopropyl-vitamin D(3) (13) is several fold more potent than the natural hormone 1alpha,25-dihydroxyvitamin D(3) (1) as an anti inflammatory agent. Here, we have further analyzed the anti-inflammatory properties of 13, confirming it as the most potent analogue tested within this family. We then determined the structures of all the natural metabolites of 13, including the 24-oxo metabolite 14, and carried out its synthesis. A comparison of 13 with 14 showed a similar induction of the primary VDR target genes CYP24A1 and CAMP and comparable anti-inflammatory properties as revealed by a similar inhibition of TNF-alpha, IL-12/23p40, IL-6, and IFN-gamma production. Interestingly, 14 displays a 3-fold lower calcemic activity in vivo compared to 13. Collectively, these findings indicate that the strong potency of 13 can be explained by the accumulation of its stable 24-oxo metabolite, which shows immunoregulatory and anti-inflammatory properties superimposable to those exerted by 13 itself. PMID- 19309156 TI - 5,7,8-Trimethyl-benzopyran and 5,7,8-trimethyl-1,4-benzoxazine aminoamide derivatives as novel antiarrhythmics against ischemia-reperfusion injury. AB - 6-Hydroxy-5,7,8-trimethyl-benzopyran derivatives and 5,7,8-trimethyl-1,4 benzoxazine analogues substituted by the lidocaine pharmacophore aminoamide functionality at C4 or N4, respectively, were synthesized and evaluated against arrhythmias associated with ischemia-reperfusion injury. The antiarrhythmic effect of substitutents at positions C2 and C6 was examined. Six out of the 11 new derivatives, exhibited arrhythmia scores 1.0-1.3 versus the control (4.5 +/- 1.2), which was also reflected to the % premature beats (0.5-3.9), control (13.7 +/- 3.6). Selected compounds were further studied by a conventional microelectrode method. 2-Diethylamino-1-(5,7,8-trimethyl-2-phenyl-2,3-dihydro benzo[1,4]oxazin-4-yl)-ethanone (50) and the trolox-inspired 4-(2-diethylamino acetyl)-2,5,7,8-tetramethyl-3,4-dihydro-2H-benzo[1,4]oxazine-2-carboxylic acid ethyl ester (62) suppress reperfusion arrhythmias and reduce malondialdehyde (MDA) content, leading to a fast recovery of the heart after ischemia reperfusion. They exhibit combined class IB and class III antiarrhythmic properties, which constitutes them as promising compounds for further studies because, due to their multichannel "amiodarone like" effect, less proarrhythmic complications can be expected. PMID- 19309157 TI - Free energy landscapes for S-H bonds in Cp*2Mo2S4 complexes. AB - An extensive family of thermochemical data is presented for a series of complexes derived from Cp*Mo(mu-S)(2)(mu-SMe)(mu-SH)MoCp* and Cp*Mo(mu-S)(2)(mu SH)(2)MoCp*. These data include electrochemical potentials, pK(a) values, homolytic solution bond dissociation free energies (SBDFEs), and hydride donor abilities in acetonitrile. Thermochemical data ranged from +0.6 to -2.0 V vs FeCp(2)(+/o) for electrochemical potentials, 5 to 31 for pK(a) values, 43 to 68 kcal/mol for homolytic SBDFEs, and 44 to 84 kcal/mol for hydride donor abilities. The observed values for these thermodynamic parameters are comparable to those of many transition metal hydrides, which is consistent with the many parallels in the chemistry of these two classes of compounds. The extensive set of thermochemical data is presented in free energy landscapes as a useful approach to visualizing and understanding the relative stabilities of all of the species under varying conditions of pH and H(2) overpressure. In addition to the previously studied homogeneous reactivity and catalysis, Mo(2)S(4) complexes are also models for heterogeneous molybdenum sulfide catalysts, and therefore, the present results demonstrate the dramatic range of S-H bond strengths available in both homogeneous and heterogeneous reaction pathways. PMID- 19309158 TI - Electrostatic control of the photoisomerization efficiency and optical properties in visual pigments: on the role of counterion quenching. AB - Hybrid QM(CASPT2//CASSCF/6-31G*)/MM(Amber) computations have been used to map the photoisomerization path of the retinal chromophore in Rhodopsin and explore the reasons behind the photoactivity efficiency and spectral control in the visual pigments. It is shown that while the electrostatic environment plays a central role in properly tuning the optical properties of the chromophore, it is also critical in biasing the ultrafast photochemical event: it controls the slope of the photoisomerization channel as well as the accessibility of the S(1)/S(0) crossing space triggering the ultrafast decay. The roles of the E113 counterion, the E181 residue, and the other amino acids of the protein pocket are explicitly analyzed: it appears that counterion quenching by the protein environment plays a key role in setting up the chromophore's optical properties and its photochemical efficiency. A unified scenario is presented that discloses the relationship between spectroscopic and mechanistic properties in rhodopsins and allows us to draw a solid mechanism for spectral tuning in color vision pigments: a tunable counterion shielding appears as the elective mechanism for L<-->M spectral modulation, while a retinal conformational control must dictate S absorption. Finally, it is suggested that this model may contribute to shed new light into mutations-related vision deficiencies that opens innovative perspectives for experimental biomolecular investigations in this field. PMID- 19309159 TI - Structure and vibrational properties of oxohalides of vanadium. AB - We study the structure and vibrational modes of a wide range of oxohalides of vanadium (VOX(n)Y(m); X, Y = F, Cl, Br, I; n, m = 0-3, n + m < or = 3). The results agree well with experimental results for VOCl(3) and VOF(3) and suggest reassignment of the experimentally observed VOF to VOF(2). We provide new assignments for various experimental modes, identifying several intermediates (VOBr(2), VOBr) and mixed structures (e.g., VOCl(2)Br), and discuss formation trends and stabilities. PMID- 19309160 TI - Ground and excited electronic states of quininone-containing Re(I)-based rectangles: a comprehensive study of their preparation, electrochemistry, and photophysics. AB - The self-assembly of two rectangular compounds [{(CO)(3)Re(mu-QL)Re(CO)(3)}(2)(mu bpy)(2)] (1, QL = 6,7-dimethyl 1,4-dioxido-9,10-anthraquinone (QL(1)); 2, QL = 1,4-dioxido-9,10-anthraquinone (QL(2)), bpy = 4,4'-bipyridine) via an orthogonal bonding approach was achieved in high yields. Their structures were characterized by single-crystal X-ray diffraction analysis. The rectangles exhibited multielectron-redox properties. The introduction of a bridging quininone moiety made notable changes in two well-separated single-electron reductions of the bpy moiety, as compared with other 2,2'-bisbenzimidazolate (BiBzlm) or thiolate- or alkoxide-bridged rectangles, followed by quasi-reversible reduction of the quininone moiety to allow the existence of different redox states. Electrochemical assessment using cyclic voltammetry and UV-vis-NIR spectroelectrochemistry revealed reversibly accessible 0, 1-, and 2- redox states. The comproportionation constant of the successive reduction processes was K(c) = 4.18 x 10(8) for complex 1 and 4.08 x 10(8) for 2. In spite of the high K(c) values, no obvious intervalence charge transfer bands were detected in either the vis, NIR, or IR regions, suggesting very weak electronic coupling between the ligand centers in the mixed-valent intermediates. In the mixed-valent intermediate, the overlap between donor and acceptor orbitals of the two bpy ligands engendered weak electronic coupling associated with distance that exceeded van der Waals ligand/ligand distances and created a class I fully isolated, non-interacting, valence-localized situation. Furthermore, unusual ligand-to-metal-to-ligand charge-transfer (LMLCT) transitions of complexes 1 and 2 at 298 K were observed in the visible region. Molecule 2 exhibited multiple emissions from the triplet-centered pi-pi* intraligand ((3)IL), metal-to-ligand charge-transfer ((3)MLCT) and triplet ligand-ligand charge transfer ((3)LLCT) levels and showed biexponential decay. By contrast, in complex 1, (3)IL emission was absent and only single-exponential decay was observed. These results reveal the different nature of the electronically excited states between 1 and 2. The mechanisms of the photophysical deactivation processes in these systems can be explained in terms of the electronic characteristics of the quininone molecules and possible geometrical differences of the excited states involved. In addition, the energies, characteristics, and molecular structures of the ground and lowest triplet excited state were calculated using the density functional theory method. PMID- 19309161 TI - An iron(II) dependent formamide hydrolase catalyzes the second step in the archaeal biosynthetic pathway to riboflavin and 7,8-didemethyl-8-hydroxy-5 deazariboflavin. AB - The early steps in the biosynthesis of 7,8-didemethyl-8-hydroxy-5-deazariboflavin (Fo) and riboflavin in the archaea differ from the established eukaryotic and bacterial pathways. The archaeal pathway has been proposed to begin with an archaeal-specific GTP cyclohydrolase III that hydrolyzes the imidazole ring of GTP but does not remove the resulting formyl group from the formamide [Graham, D. E., Xu, H., and White, R. H. (2002) Biochemistry 41, 15074-15084 ]. This enzyme is different than the bacterial GTP cyclohydrolase II which catalyzes both reactions. Here we describe the identification and characterization of the formamide hydrolase that catalyzes the second step in the archaeal Fo and riboflavin biosynthetic pathway. The Methanocaldococcus jannaschii MJ0116 gene was cloned and heterologously expressed, and the resulting enzyme was shown to catalyze the formation of 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5' phosphate (APy) and formate from 2-amino-5-formylamino-6-ribosylamino-4(3H) pyrimidinone 5'-monophosphate (FAPy). The MJ0116-derived protein has been named ArfB to indicate that it catalyzes the second step in archaeal riboflavin and Fo biosynthesis. ArfB was found to require ferrous iron for activity although metal analysis by ICP indicated the presence of zinc as well as iron in the purified protein. The identification of this enzyme confirms the involvement of GTP cyclohydrolase III (ArfA) in archaeal riboflavin and Fo biosynthesis. PMID- 19309162 TI - Conantokin-Br from Conus brettinghami and selectivity determinants for the NR2D subunit of the NMDA receptor. AB - Conantokins are venom peptides from marine cone snails that are NMDA receptor antagonists. Here, we report the characterization of a 24 AA conantokin from Conus brettinghami Coomans , H. E. , Moolenbeek , R. G. and Wils , E. ( 1982 ) Basteria 46 ( 1/4 ), 3 - 67 , conantokin-Br (con-Br), the first conantokin that does not have the conserved glutamate residue at position 2. Molecular modeling studies suggest that con-Br has a helical structure between residues 2-13. In contrast to other characterized conantokins, con-Br has a high potency for NMDA receptors with NR2D subunits. To identify determinants for NR2D potency, we synthesized chimeras of con-Br and conantokin-R (con-R); the latter has a approximately 30-fold lower potency for the NR2D subtype. The characterization of two reciprocal chimeras (con-Br/R and con-R/Br), comprising the first 9-10 N terminal AAs of each conantokin followed by the corresponding C-terminal AAs of the other conantokin demonstrates that determinants for NR2D selectivity are at the N-terminal region. Additional analogues comprising 1-3 amino acid substitutions from each peptide into the homologous region of the other led to the identification of a key determinant; a Tyr residue in position 5 increases potency for NR2D, while Val at this locus causes a decrease. The systematic definition of key determinants in the conantokin peptides for NMDA receptor subtype selectivity is an essential component in the development of conantokin peptides that are highly selective for each specific NMDA receptor subtype. PMID- 19309163 TI - Oxidative folding pathway of onconase, a ribonuclease homologue: insight into oxidative folding mechanisms from a study of two homologues. AB - The oxidative folding pathways of two four-disulfide proteins of the ribonuclease family, ONC and RNase A, which have similar three-dimensional folds but only 30% sequence homology, are compared. In this study, a mechanism for the oxidative folding pathway of ONC is proposed. In particular, the kinetic roles and thermodynamic characteristics of key intermediates along the oxidative folding pathway, specifically, the structured intermediates, I(1), I(2), and I(3), previously identified as des-[19-68,30-75], des-[30-75], and des-[19-68], respectively, are discussed. In addition, the effects of temperature on the oxidative folding pathway have been examined. Differences in the folding mechanism between ONC and RNase A are attributed to the differences in their amino acid sequences and related inter-residue interactions, including differences in hydrophobic interactions. Compared to RNase A, ONC utilizes more efficient interactions along the oxidative folding pathway to adopt its native fold more rapidly. PMID- 19309164 TI - Bridging macro and nano. PMID- 19309166 TI - The ring: a leitmotif in plasmonics. AB - An interesting paper in this issue describes an experiment that is an optical analogue of the "quantum corral" on metal surfaces constructed and observed using scanning tunneling microscopy more than a decade ago. In this Perspective, the mechanism underlying the confinement and manipulation of light in metallic nanorings and related structures are discussed, with an emphasis on recent results. PMID- 19309167 TI - Micro-mirrors for nanoscale three-dimensional microscopy. AB - A research-grade optical microscope is capable of resolving fine structures in two-dimensional images. However, three-dimensional resolution, or the ability of the microscope to distinguish between objects lying above or below the focal plane from in-focus objects, is not nearly as good as in-plane resolution. In this issue of ACS Nano, McMahon et al. report the use of mirrored pyramidal wells with a conventional microscope for rapid, 3D localization and tracking of nanoparticles. Mirrors have been used in microscopy before, but recent work with MPWs is unique because it enables the rapid determination of the x-, y-, and z position of freely diffusing nanoparticles and cellular nanostructures with unprecedented speed and spatial accuracy. As inexpensive tools for 3D visualization, mirrored pyramidal wells may prove to be invaluable aids in nanotechnology and engineering of nanomaterials. PMID- 19309168 TI - Bend, buckle, and fold: mechanical engineering with nanomembranes. AB - Research on nanomembranes and graphene sheets represents the "third wave" of work on nanomaterials, following earlier studies of nanoparticles/fullerenes and, somewhat later, nanowires/nanotubes. Inorganic semiconductor nanomembranes are particularly appealing due to their materials diversity, the ease with which they can be grown with high quality over large areas, and the ability to exploit them in unique, high-performance electronic and optoelectronic systems. The mechanics of such nanomembranes and the coupling of strain to their electronic properties are topics of considerable current interest. A new paper by the Lagally group in this issue combines single-crystalline silicon nanomembranes with chemical vapor deposition techniques to form "mechano-electronic" superlattices whose properties could lead to unusual classes of electronic devices. PMID- 19309169 TI - Observation of all the intermediate steps of a chemical reaction on an oxide surface by scanning tunneling microscopy. AB - By means of high-resolution scanning tunneling microscopy (STM), we have revealed unprecedented details about the intermediate steps for a surface-catalyzed reaction. Specifically, we studied the oxidation of H adatoms by O(2) molecules on the rutile TiO(2)(110) surface. O(2) adsorbs and successively reacts with the H adatoms, resulting in the formation of water species. Using time-lapsed STM imaging, we have unraveled the individual reaction intermediates of HO(2), H(2)O(2), and H(3)O(2) stoichiometry and the final reaction product-pairs of water molecules, [H(2)O](2). Because of their different appearance and mobility, these four species are discernible in the time-lapsed STM images. The interpretation of the STM results is corroborated by density functional theory calculations. The presented experimental and theoretical results are discussed with respect to previous reports where other reaction mechanisms have been put forward. PMID- 19309170 TI - Cell electrophysiology with carbon nanopipettes. AB - The ability to monitor living cell behavior in real time and with high spatial resolution is vital for advancing our knowledge of cellular machinery and evaluating cellular response to various drugs. Here, we report the development and utilization of carbon-based nanoelectrodes for cell electrophysiology. We employ carbon nanopipettes (CNPs), novel carbon-based nanoprobes which integrate carbon nanopipes into the tips of pulled glass capillaries, to measure electrical signals in the mouse hippocampal cell line HT-22. Using a standard electrophysiology amplifier in current-clamp mode, we measured the resting membrane potential of cells and their transient membrane response to extracellular pharmacological agents. In addition to their superior injection capabilities reported previously, CNPs are capable of multifunctionality, enabling, for example, concurrent intracellular injection and electrical measurements without damaging cells. PMID- 19309171 TI - 3D particle trajectories observed by orthogonal tracking microscopy. AB - We demonstrate high-resolution, high-speed 3D nanoparticle tracking using angled micromirrors. When angled micromirrors are introduced into the field of view of an optical microscope, reflected side-on views of a diffusing nanoparticle are projected alongside the usual direct image. The experimental design allows us to find the 3D particle trajectory using fast, centroid-based image processing, with no nonlinear computing operations. We have tracked polystyrene particles of 190 nm diameter with position measurement precision <20 nm in 3D with 3 ms frame duration (i.e., at an imaging rate >330 frames per second). Because the image processing requires only approximately 1 ms per frame, this technique could enable real-time feedback-controlled nanoparticle assembly applications with nanometer precision. PMID- 19309172 TI - Tunability of subradiant dipolar and fano-type plasmon resonances in metallic ring/disk cavities: implications for nanoscale optical sensing. AB - Plasmonic nanocavities consisting of the concentric arrangement of a disk and a ring sustain both subradiant and superradiant dipolar plasmon modes with large associated field enhancements and high refractive index sensitivities. In structures with broken symmetry, additionally a highly tunable Fano interference feature appears, which can be explained with a simple analytical harmonic oscillator model. The spectral tunability of these resonances from the visible to the mid-infrared is investigated, highlighting a potential for applications in surface enhanced spectroscopies. PMID- 19309173 TI - Benzaldehyde-functionalized polymer vesicles. AB - Polymer vesicles with diameters of ca. 100-600 nm and bearing benzaldehyde functionalities within the vesicular walls were constructed through self-assembly of an amphiphilic block copolymer PEO(45)-b-PVBA(26) in water. The reactivity of the benzaldehyde functionalities was verified by cross-linking the polymersomes and also by a one-pot cross-linking and functionalization approach to further render the vesicles fluorescent, each via reductive amination. In vitro studies found these labeled nanostructures to undergo cell association. PMID- 19309174 TI - Highly efficient quantum-dot light-emitting diodes with DNA-CTMA as a combined hole-transporting and electron-blocking layer. AB - Owing to their narrow bright emission band, broad size-tunable emission wavelength, superior photostability, and excellent flexible-substrate compatibility, light-emitting diodes based on quantum dots (QD-LEDs) are currently under intensive research and development for multiple consumer applications including flat-panel displays and flat lighting. However, their commercialization is still precluded by the slow development to date of efficient QD-LEDs as even the highest reported efficiency of 2.0% cannot favorably compete with their organic counterparts. Here, we report QD-LEDs with a record high efficiency (approximately 4%), high brightness (approximately 6580 cd/m(2)), low turn-on voltage (approximately 2.6 V), and significantly improved color purity by simply using deoxyribonucleic acid (DNA) complexed with cetyltrimetylammonium (CTMA) (DNA-CTMA) as a combined hole transporting and electron-blocking layer (HTL/EBL). This, together with controlled thermal decomposition of ligand molecules from the QD shell, represents a novel combined, but simple and very effective, approach toward the development of highly efficient QD-LEDs with a high color purity. PMID- 19309175 TI - Software platform for rapidly creating computational tools for mass spectrometry based proteomics. AB - We describe and demonstrate the proteomics computational toolkit provided in the open-source msInspect software distribution. The toolkit includes modules written in Java and in the R statistical programming language to aid the rapid development of proteomics software applications. It contains tools for processing and manipulating standard MS data files, including signal processing of LC-MS data and parsing of MS/MS search results, as well as for modeling proteomics data structures, creating charts, and other common tasks. We present this toolkit's capability to rapidly develop new computational tools by presenting an example application, Qurate, a graphical tool for manually curating isotopically labeled peptide quantitative events. PMID- 19309176 TI - Prediction of HPLC retention index using artificial neural networks and IGroup E state indices. AB - A back-propagation artificial neural network (ANN) was used to create a 10-fold leave-10%-out cross-validated ensemble model of high performance liquid chromatography retention index (HPLC-RI) for a data set of 498 diverse druglike compounds. A 10-fold multiple linear regression (MLR) ensemble model of the same data was developed for comparison. Molecular structure was described using IGroup E-state indices, a novel set of structure-information representation (SIR) descriptors, along with molecular connectivity chi and kappa indices and other SIR descriptors previously reported. The same input descriptors were used to develop models by both learning algorithms. The MLR model yielded marginally acceptable statistics with training correlation r(2) = 0.65, mean absolute error (MAE) = 83 RI units. External validation of 104 compounds not used for model development yielded validation v(2) = 0.49 and MAE = 73 RI units. The distribution of residuals for the fit and validate data sets suggest a nonlinear relationship between retention index and molecular structure as described by the SIR indices. Not surprisingly, the ANN model was significantly more accurate for both training and validation with training set r(2) = 0.93, MAE = 30 RI units and validation v(2) = 0.84, MAE = 41 RI units. For the ANN model, a total of 91% of validation predictions were within 100 RI units of the experimental value. PMID- 19309177 TI - Efficient exploration of large combinatorial chemistry spaces by monomer-based similarity searching. AB - In modern drug discovery, 2-D similarity searching is widely employed as a cost effective way to screen large compound collections and select subsets of molecules that may have interesting biological activity prior to experimental screening. Nowadays, there is a growing interest in applying the existing 2-D similarity searching methods to combinatorial chemistry libraries to search for novel hits or to evolve lead series. A dilemma thus arises when many identical substructures recur in library products and they have to be considered repeatedly in descriptor calculations. The dilemma is exacerbated by the astronomical number of combinatorial products. This problem imposes a major barrier to similarity searching of large combinatorial chemistry spaces. An efficient approach, termed Monomer-based Similarity Searching (MoBSS), is proposed to remedy the problem. MoBSS calculates atom pair (AP) descriptors based on interatomic topological distances, which lend themselves to pair additivity. A fast algorithm is employed in MoBSS to rapidly compute product atom pairs from those of the constituent fragments. The details of the algorithm are presented along with a series of proof-of-concept studies, which demonstrate the speed, accuracy, and utility of the MoBSS approach. PMID- 19309178 TI - Partition behavior of polycyclic aromatic hydrocarbons between aged coal tar and water. AB - Coal tar aged in a large-scale, artificial aquifer experiment for five years was subsequently investigated for leaching behavior of polycyclic aromatic hydrocarbons (PAHs). After five years, the initially liquid coal tar had solidified and formed segregated particles with a grain size similar to that of the sandy aquifer material. The composition of the aged coal tar (ACT) with regard to PAHs was remarkably different from that of the original bulk coal tar (BCT), because most of the low-molecular-weight compounds had been depleted. Equilibrium aqueous-phase concentrations of 17 PAHs leaching from the aquifer material containing the ACT were measured from consecutive equilibration steps at increasing temperatures of between 25 and 100 degrees C using accelerated solvent extraction. The results showed 2- to 5,000-fold lower concentrations than those from BCT, indicating dramatic changes of dissolution behavior of PAHs from coal tar after the five-year aging period. Predictions based on Raoult's law with the subcooled liquid solubilities substantially overestimated the equilibrium aqueous phase concentrations of the PAHs from ACT, whereas the estimations were reasonable if the solid solubilities were employed instead. The enthalpies of phase transfer from ACT to water were determined based on the van't Hoff equation. The resulting values agreed with the dissolution enthalpies of pure solid rather than subcooled liquid PAHs. PMID- 19309179 TI - Impact of kerogen heterogeneity on sorption of organic pollutants. 2. Sorption equilibria. AB - Phenanthrene and naphthalene sorption isotherms were measured for three different series of kerogen materials using completely mixed batch reactors. Sorption isotherms were nonlinear for each sorbate-sorbent system, and the Freundlich isotherm equation fit the sorption data well. The Freundlich isotherm linearity parameter n ranged from 0.192 to 0.729 for phenanthrene and from 0.389 to 0.731 for naphthalene. The n values correlated linearly with rigidity and aromaticity of the kerogen matrix, but the single-point, organic carbon-normalized distribution coefficients varied dramatically among the tested sorbents. A dual mode sorption equation consisting of a linear partitioning domain and a Langmuir adsorption domain adequately quantified the overall sorption equilibrium for each sorbent-sorbate system. Both models fit the data well, with r2 values of 0.965 to 0.996 for the Freundlich model and 0.963 to 0.997 for the dual-mode model for the phenanthrene sorption isotherms. The dual-mode model fitting results showed that as the rigidity and aromaticity of the kerogen matrix increased, the contribution of the linear partitioning domain to the overall sorption equilibrium decreased, whereas the contribution of the Langmuir adsorption domain increased. The present study suggested that kerogen materials found in soils and sediments should not be treated as a single, unified, carbonaceous sorbent phase. PMID- 19309180 TI - Rates of change in naturalistic psychotherapy: contrasting dose-effect and good enough level models of change. AB - Most research on the dose-effect model of change has combined data across patients who vary in their total dose of treatment and has implicitly assumed that the rate of change during therapy is constant across doses. In contrast, the good-enough level model predicts that rate of change will be related to total dose of therapy. In this study, the authors evaluated these competing predictions by examining the relationship between rate of change and total dose in 4,676 psychotherapy patients who received individual psychotherapy. Patients attended 6.46 sessions on average (SD = 4.14, range = 3-29, Mdn = 5). The results indicated that although patients improved during treatment, patients' rate of change varied as a function of total dose of treatment. Small doses of treatment were related to relatively fast rates of change, whereas large doses of treatment were related to slower rates of change. Total dose had a nonlinear relationship with the likelihood of clinically significant change. Given the variability in rates of change, it appears that time limits for treatment uniform to all patients would not adequately serve patients' needs. PMID- 19309181 TI - Barriers to implementing treatment integrity procedures: survey of treatment outcome researchers. AB - Treatment integrity refers to implementing interventions as intended. Treatment integrity is critically important for experimental validity and for drawing valid inferences regarding the relationship between treatment and outcome. Yet, it is rarely adequately addressed in psychotherapy research. The authors examined barriers to treatment integrity implementation by surveying psychotherapy researchers. Results indicate that lack of theory and guidelines on treatment integrity procedures, as well as time, cost, and labor constraints, were regarded as strong barriers. The lack of general knowledge about treatment integrity and the lack of editorial requirement for reporting integrity procedures were also perceived as barriers to its implementation. However, psychotherapy researchers indicated awareness of the importance of treatment integrity for the experimental validity of a study and did not regard lack of its appreciation as a barrier for implementing integrity procedures. Further, a higher number of endorsed barriers predicted lower adequacy of treatment integrity procedures in the authors' own research. Recommendations for improving how integrity is addressed include journal and editorial enforcement of treatment integrity implementation, funding for integrity procedures, and provision of specific guidelines. PMID- 19309182 TI - A behavioral economic reward index predicts drinking resolutions: moderation revisited and compared with other outcomes. AB - Data were pooled from 3 studies of recently resolved community-dwelling problem drinkers to determine whether a behavioral economic index of the value of rewards available over different time horizons distinguished among moderation (n = 30), abstinent (n = 95), and unresolved (n = 77) outcomes. Moderation over 1- to 2 year prospective follow-up intervals was hypothesized to involve longer term behavior regulation processes than abstinence or relapse and to be predicted by more balanced preresolution monetary allocations between short-term and longer term objectives (i.e., drinking and saving for the future). Standardized odds ratios (ORs) based on changes in standard deviation units from a multinomial logistic regression indicated that increases on this "Alcohol-Savings Discretionary Expenditure" index predicted higher rates of abstinence (OR = 1.93, p = .004) and relapse (OR = 2.89, p < .0001) compared with moderation outcomes. The index had incremental utility in predicting moderation in complex models that included other established predictors. The study adds to evidence supporting a behavioral economic analysis of drinking resolutions and shows that a systematic analysis of preresolution spending patterns aids in predicting moderation. PMID- 19309183 TI - Changing network support for drinking: network support project 2-year follow-up. AB - The Network Support Project was designed to determine whether a treatment could lead patients to change their social network from one that supports drinking to one that supports sobriety. This study reports 2-year posttreatment outcomes. Alcohol-dependent men and women (N = 210) were randomly assigned to 1 of 3 outpatient treatment conditions: network support (NS), network support + contingency management (NS + CM), or case management (CaseM, a control condition). Analysis of drinking rates indicated that the NS condition yielded up to 20% more days abstinent than the other conditions at 2 years posttreatment. NS treatment also resulted in greater increases at 15 months in social network support for abstinence, as well as in AA attendance and AA involvement than did the other conditions. Latent growth modeling suggested that social network changes were accompanied by increases in self-efficacy and coping that were strongly predictive of long-term drinking outcomes. The findings indicate that a network support treatment can effect long-term adaptive changes in drinkers' social networks and that these changes contribute to improved drinking outcomes in the long term. PMID- 19309184 TI - A randomized trial of individual and couple behavioral alcohol treatment for women. AB - Although alcohol use disorders (AUDs) adversely affect women, research on efficacious treatments for women is limited. In this randomized efficacy trial of 102 heterosexual women with AUDs, the authors compared alcohol behavioral couple therapy (ABCT) and alcohol behavioral individual therapy (ABIT) on percentage of days abstinent (PDA) and percentage of days of heavy drinking (PDH) over 6 months of treatment and 12 months of posttreatment follow-up. Baseline relationship functioning and comorbid disorders were tested as moderators of outcome. Piecewise linear growth models were used to model outcomes. During treatment, women increased their PDA and decreased their PDH, with significantly greater improvements in ABCT than in ABIT (d = 0.59 for PDA; d = 0.79 for PDH). Differences favoring ABCT were maintained during follow-up. Women with poorer baseline relationship functioning improved more on PDA during treatment with ABCT than with ABIT. For PDH, results during treatment and follow-up favored ABCT for women with better baseline relationship functioning. ABCT resulted in better outcomes than ABIT for women with Axis I disorders at the end of follow-up (PDA), and for women with Axis II disorders at the end of treatment (PDA) and at the end of follow-up (PDH). PMID- 19309185 TI - A practical clinical trial of coordinated care management to treat substance use disorders among public assistance beneficiaries. AB - This study tested whether coordinated care management (CCM), a continuity of care intervention for substance use disorders (SUD), improved rates of abstinence when compared with usual welfare management for substance-using single adults and adults with dependent children applying for public assistance. The study was designed as a practical clinical trial and was implemented in partnership with a large city welfare agency. Participants were 421 welfare applicants identified via SUD screening and assigned via an unbiased computerized allocation program to a site that provided either CCM (n = 232) or usual care (UC; n = 189). Outcomes were assessed for 1 year postbaseline with self-reports and biological measures of substance use. As hypothesized, for participants not enrolled in methadone maintenance programs (n = 313), CCM clients received significantly more services than did UC clients. Nonmethadone CCM also showed significantly higher abstinence rates (odds ratio = 1.75; 95% confidence interval = 1.12, 2.76; d = 0.31) that emerged early in treatment and were sustained throughout follow-up. In contrast, no treatment services or outcome effects were found for methadone maintenance clients (n = 108). Findings suggest that CCM is promising as a wraparound to SUD treatment for welfare recipients. PMID- 19309186 TI - The impact of evidence-based practice implementation and fidelity monitoring on staff turnover: evidence for a protective effect. AB - Staff retention is an ongoing challenge in mental health and community-based service organizations. Little is known about the impact of evidence-based practice implementation on the mental health and social service workforce. The present study examined the effect of evidence-based practice implementation and ongoing fidelity monitoring on staff retention in a children's services system. The study took place in the context of a statewide, regionally randomized effectiveness trial of an evidence-based intervention designed to reduce child neglect. In the study 21 teams consisting of 153 home-based service providers were followed over a 29-month period. Survival analyses revealed greater staff retention in the condition where the evidence-based practice was implemented along with ongoing fidelity monitoring presented to staff as supportive consultation. These results should help to allay concerns about staff retention when implementing evidence-based practices where there is good values-innovation fit and when fidelity monitoring is designed as an aid and support to service providers in providing a high standard of care for children and families. PMID- 19309187 TI - Psychiatric diagnoses as contemporaneous risk factors for suicide attempts among adolescents and young adults: developmental changes. AB - The purpose of this prospective, naturalistic study was to examine the relationships between suicide attempts and contemporaneous psychiatric disorders, and developmental changes in these relationships from adolescence to young adulthood. The sample consisted of 180 adolescents, 12-19 years of age at hospitalization, repeatedly assessed for up to 13 years (n = 1,825 assessments). Semistructured psychiatric diagnostic instruments were administered at repeated assessments to assess psychiatric disorders and suicide attempts. After controlling for demographic variables and prehospitalization suicide attempts, most contemporaneous psychiatric disorders (major depressive disorder [MDD], dysthymic disorder, generalized anxiety disorder [GAD], panic disorder, attention deficit/hyperactivity disorder [AD/HD], conduct disorder, and substance use disorder [SUD]) were related to increased risk of attempts. The relationship between suicide attempts and MDD, GAD, AD/HD, and SUD strengthened as participants got older. MDD, dysthymic disorder, GAD, and panic disorder were more commonly associated with repeat than 1st-time suicide attempts. In sum, most major psychiatric disorders are associated with increased risk for suicide attempts, but the strength of the relationships between these disorders and attempts changes over the course of development. PMID- 19309188 TI - Supporting affect regulation in children with multiple disabilities during psychotherapy: a multiple case design study of therapeutic attachment. AB - In a controlled multiple case design study, the development of a therapeutic relationship and its role in affect regulation were studied in 6 children with visual disabilities, severe intellectual disabilities, severe challenging behavior, and prolonged social deprivation. In the 1st phase, children had sessions with an experimental therapist stimulating therapeutic attachment, alternating with a control therapist providing positive personal attention only. In the 2nd phase, both therapists applied behavior therapy. Clients sought more proximity to the experimental therapist compared with the control therapist. Psychophysiological arousal (respiratory sinus arrhythmia and pre-ejection period) was lower when the experimental therapist applied behavior modification than when the control therapist did. Despite prolonged social deprivation, the attachment behavioral system appeared responsive to stimulation. The effects on affect regulation may explain the synergy between psychotherapy based on interpersonal and behavior modification approaches. PMID- 19309189 TI - Targeting children's behavior problems in preschool classrooms: a cluster randomized controlled trial. AB - The present study evaluated the efficacy of a multicomponent, classroom-based intervention in reducing preschoolers' behavior problems. The Chicago School Readiness Project model was implemented in 35 Head Start classrooms using a clustered-randomized controlled trial design. Results indicate significant treatment effects (ds = 0.53-0.89) for teacher-reported and independent observations of children's internalizing and externalizing behavior problems. Moreover, there was some evidence for the moderating role of child gender, race/ethnic group membership, and exposure to poverty-related risk, with stronger effects of intervention for some groups of children than for others. Findings contribute to a growing area of research on poverty and preventive intervention in early childhood. PMID- 19309190 TI - Testing the efficacy of theoretically derived improvements in the treatment of social phobia. AB - Recent theoretical models of social phobia suggest that targeting several specific cognitive factors in treatment should enhance treatment efficacy over that of more traditional skills-based treatment programs. In the current study, 195 people with social phobia were randomly allocated to 1 of 3 treatments: standard cognitive restructuring plus in vivo exposure, an "enhanced" treatment that augmented the standard program with several additional treatment techniques (e.g., performance feedback, attention retraining), and a nonspecific (stress management) treatment. The enhanced treatment demonstrated significantly greater effects on diagnoses, diagnostic severity, and anxiety during a speech. The specific treatments failed to differ significantly on self-report measures of social anxiety symptoms and life interference, although they were both significantly better than the nonspecific treatment. The enhanced treatment also showed significantly greater effects than standard treatment on 2 putative process measures: cost of negative evaluation and negative views of one's skills and appearance. Changes on these process variables mediated differences between the treatments on changes in diagnostic severity. PMID- 19309191 TI - Therapeutic responses of psychopathic sexual offenders: treatment attrition, therapeutic change, and long-term recidivism. AB - The authors examined the therapeutic responses of psychopathic sex offenders (>or=25 Psychopathy Checklist--Revised; PCL-R) in terms of treatment dropout and therapeutic change, as well as sexual and violent recidivism over a 10-year follow-up among 156 federally incarcerated sex offenders treated in a high intensity inpatient sex offender program. Psychopathy and sex offender risk/treatment change were assessed using the PCL-R and the Violence Risk Scale- Sexual Offender version (VRS-SO), respectively. Although psychopathic participants were more likely than their nonpsychopathic counterparts (<25 PCL-R) to drop out, almost 75% of the former completed treatment. Psychopathic offenders who failed to complete sex offender treatment were more likely to violently but not sexually recidivate than completers. Positive treatment changes were associated with reductions in sexual and violent recidivism after psychopathy and sexual recidivism risk were controlled. Overall, the results suggest that given appropriate treatment interventions, sex offenders with significant psychopathic traits can be retained in an institutional treatment program and those showing therapeutic improvement can reduce their risk for both sexual and violent recidivism. PMID- 19309193 TI - Gender differences in structured risk assessment: comparing the accuracy of five instruments. AB - Structured risk assessment should guide clinical risk management, but it is uncertain which instrument has the highest predictive accuracy among men and women. In the present study, the authors compared the Psychopathy Checklist Revised (PCL-R; R. D. Hare, 1991, 2003); the Historical, Clinical, Risk Management-20 (HCR-20; C. D. Webster, K. S. Douglas, D. Eaves, & S. D. Hart, 1997); the Risk Matrix 2000-Violence (RM2000[V]; D. Thornton et al., 2003); the Violence Risk Appraisal Guide (VRAG; V. L. Quinsey, G. T. Harris, M. E. Rice, & C. A. Cormier, 1998); the Offenders Group Reconviction Scale (OGRS; J. B. Copas & P. Marshall, 1998; R. Taylor, 1999); and the total previous convictions among prisoners, prospectively assessed prerelease. The authors compared predischarge measures with subsequent offending and instruments ranked using multivariate regression. Most instruments demonstrated significant but moderate predictive ability. The OGRS ranked highest for violence among men, and the PCL-R and HCR-20 H subscale ranked highest for violence among women. The OGRS and total previous acquisitive convictions demonstrated greatest accuracy in predicting acquisitive offending among men and women. Actuarial instruments requiring no training to administer performed as well as personality assessment and structured risk assessment and were superior among men for violence. PMID- 19309194 TI - Predicting attention-deficit/hyperactivity disorder and oppositional defiant disorder from preschool diagnostic assessments. AB - The present study examined the power of measures of early preschool behavior to predict later diagnoses of attention-deficit/hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD)/conduct disorder (CD). Participants were 168 children with behavior problems at age 3 who underwent a multimethod assessment of ADHD and ODD symptoms and were followed annually for 3 years. Fifty-eight percent of 3-year-old children with behavior problems met criteria for ADHD and/or ODD/CD 3 years later. Using a diagnostic interview and rating scales at age 3, the authors could accurately predict later diagnostic status for 3/4 of children for ADHD and for 2/3 of children for ODD/CD. Predictive power of the best models did not increase significantly at age 4 and age 5 compared with age 3. Results provide support for the validity of early diagnoses of ADHD, although caution is needed in making diagnoses because a significant minority of children with early hyperactivity and inattention do outgrow their problems. PMID- 19309195 TI - Decreased family accommodation associated with improved therapy outcome in pediatric obsessive-compulsive disorder. AB - Pediatric obsessive-compulsive disorder (OCD) is a chronic, disabling condition that affects both patients and their families. Despite the identification of efficacious treatments (e.g., cognitive-behavioral therapy and selective serotonin reuptake inhibitor medications), not all patients respond fully. The purpose of the present study was to examine whether the amount of family accommodation provided to pediatric patients with OCD is associated with treatment outcome, and whether decreases in accommodation are associated with improved outcome. The sample consisted of 49 youths (6-18 years of age), who participated in 14 sessions of family-based cognitive-behavioral therapy for OCD, and their parents. Participants completed measures at pretreatment and posttreatment. Results indicate that family accommodation was prevalent among families of pediatric patients with OCD and that such accommodation was associated with symptom severity at pretreatment. In addition, decreases in family accommodation during treatment predicted treatment outcome, even when controlling for pretreatment OCD severity-impairment. Results suggest that the level of accommodation provided by the family may indicate an important obstacle to, or predictor of, treatment outcome in pediatric OCD. Directions for future research are discussed. PMID- 19309196 TI - Prediction of sexual assault experiences in college women based on rape scripts: a prospective analysis. AB - Although script theory has been applied to sexual assault (e.g., H. Frith & C. Kitzinger, 2001; A. S. Kahn, V. A. Andreoli Mathie, & C. Torgler, 1994), women's scripts of rape have not been examined in relation to predicting sexual victimization experiences. The purpose of the current study was to examine how elements of women's sexual assault scripts predicted their sexual assault experiences over a follow-up period. The authors used data from a baseline and follow-up session for 339 undergraduate women. The results suggest that women who constructed narratives containing certain elements were more likely to report a sexual assault over the academic quarter. Specifically, narratives containing the woman utilizing nonforceful resistance, the woman having less control over the outcome of the situation, the assault happening outdoors, the assault being more severe, and the woman having known the perpetrator less time were predictive of reported sexual victimization over the 8-week follow-up period. Additionally, having a history of adolescent sexual victimization was also predictive of reported sexual victimization over the quarter. These findings have important implications in sexual assault risk-reduction programming, which are discussed. PMID- 19309197 TI - Continuation-phase cognitive therapy's effects on remission and recovery from depression. AB - The authors tested the effects of continuation-phase cognitive therapy (C-CT) on remission and recovery from recurrent major depressive disorder, defined as 6 weeks and 8 months, respectively, of continuously absent or minimal symptoms. Responders to acute-phase cognitive therapy were randomized to 8 months of C-CT (n = 41) or assessment control (n = 43), and they were followed 16 additional months (R. B. Jarrett et al., 2001). Relative to controls, a few more patients in C-CT remitted (88% vs. 97%), and significantly more recovered (62% vs. 84%). All patients without remission and recovery relapsed, but most patients who remitted (60%) and who recovered (75%) did not later relapse or recur. The authors discuss the importance of defining efficacious treatment as producing remission and recovery. PMID- 19309198 TI - Social exclusion and early-stage interpersonal perception: selective attention to signs of acceptance. AB - Social exclusion can thwart people's powerful need for social belonging. Whereas prior studies have focused primarily on how social exclusion influences complex and cognitively downstream social outcomes (e.g., memory, overt social judgments and behavior), the current research examined basic, early-in-the-cognitive-stream consequences of exclusion. Across 4 experiments, the threat of exclusion increased selective attention to smiling faces, reflecting an attunement to signs of social acceptance. Compared with nonexcluded participants, participants who experienced the threat of exclusion were faster to identify smiling faces within a "crowd" of discrepant faces (Experiment 1), fixated more of their attention on smiling faces in eye-tracking tasks (Experiments 2 and 3), and were slower to disengage their attention from smiling faces in a visual cueing experiment (Experiment 4). These attentional attunements were specific to positive, social targets. Excluded participants did not show heightened attention to faces conveying social disapproval or to positive nonsocial images. The threat of social exclusion motivates people to connect with sources of acceptance, which is manifested not only in "downstream" choices and behaviors but also at the level of basic, early-stage perceptual processing. PMID- 19309199 TI - Cultural sharing in a global village: evidence for extracultural cognition in European Americans. AB - The authors examined the effects of exposure to foreign cultural environments and symbols on decision making among European Americans. Although European Americans predicted change less frequently than East Asians did (Pilot Study A), European Americans anticipated greater change when primed with East Asian culturally-laden locations (Pilot Study B and Study 1) and the East Asian yin-yang symbol (Studies 2-7). These effects held in the domains of stock prediction and weather forecasting and were stronger the more familiar European Americans were with the cultural primes, and the longer they had spent overseas. Together, these findings suggest that familiar culturally-laden cues sometimes prime people within one cultural milieu to make so-called extracultural judgments. PMID- 19309200 TI - Looking into the past: cultural differences in perception and representation of past information. AB - The authors investigated cultural differences in the way people perceive and represent temporal information. It was hypothesized that Chinese would attend to the past information more than would Canadians. In Studies 1 and 2, Canadian and Chinese participants read a description of a theft along with a list of behaviors that occurred in the past or present. Chinese participants rated behaviors that had taken place in the remote and recent past as more relevant to solving the case than did Canadians. Study 3 showed that Chinese participants recalled greater detail about past events than did Canadians. Studies 4A and 4B showed that Chinese perceived past events as being closer to the present than did Canadians, suggesting that Chinese had a greater awareness of the past. Overall, Chinese attended to a greater range of past information than did Canadians, which has significant theoretical and practical implications. PMID- 19309201 TI - Self-affirmation and self-control: affirming core values counteracts ego depletion. AB - Research has established that acts of self-control deplete a resource required for subsequent self-control tasks. The present investigation revealed that a psychological intervention-self-affirmation-facilitates self-control when the resource has been depleted. Experiments 1 and 2 found beneficial effects of self affirmation on self-control in a depleted state. Experiments 3 and 4 suggested that self-affirmation improves self-control by promoting higher levels (vs. lower levels) of mental construal. Self-affirmation therefore holds promise as a mental strategy that reduces the likelihood of self-control failure. PMID- 19309202 TI - A mechanistic explanation of popularity: genes, rule breaking, and evocative gene environment correlations. AB - Previous work has suggested that the serotonergic system plays a key role in "popularity" or likeability. A polymorphism within the 5HT-sub(2A) serotonin receptor gene (-G1438A) has also been associated with popularity, suggesting that genes may predispose individuals to particular social experiences. However, because genes cannot code directly for others' reactions, any legitimate association should be mediated via the individual's behavior (i.e., genes- >behaviors-->social consequences), a phenomenon referred to as an evocative gene environment correlation (rGE). The current study aimed to identify one such mediating behavior. The author focused on rule breaking given its prior links to both the serotonergic system and to increased popularity during adolescence. Two samples of previously unacquainted late-adolescent boys completed a peer-based interaction paradigm designed to assess their popularity. Analyses revealed that rule breaking partially mediated the genetic effect on popularity, thereby furthering our understanding of the biological mechanisms that underlie popularity. Moreover, the present results represent the first meaningfully explicated evidence that genes predispose individuals not only to particular behaviors but also to the social consequences of those behaviors. PMID- 19309203 TI - Not so black and white: memory for ambiguous group members. AB - Exponential increases in multiracial identities, expected over the next century, create a conundrum for perceivers accustomed to classifying people as their own- or other-race. The current research examines how perceivers resolve this dilemma with regard to the own-race bias. The authors hypothesized that perceivers are not motivated to include ambiguous-race individuals in the in-group and therefore have some difficulty remembering these individuals. Both racially ambiguous and other-race faces were misremembered more often than own-race faces (Study 1), though memory for ambiguous faces was improved among perceivers motivated to include biracial individuals in the in-group (Study 2). Racial labels assigned to racially ambiguous faces determined memory for these faces, suggesting that uncertainty provides the motivational context for discounting ambiguous faces in memory (Study 3). Finally, an inclusion motivation fostered cognitive associations between racially ambiguous faces and the in-group. Moreover, the extent to which perceivers associated racially ambiguous faces with the in-group predicted memory for ambiguous faces and accounted for the impact of motivation on memory (Study 4). Thus, memory for biracial individuals seems to involve a flexible person construal process shaped by motivational factors. PMID- 19309204 TI - When trying to understand detracts from trying to behave: effects of perspective taking in intergroup interaction. AB - Four studies demonstrate that perspective taking can backfire in intergroup interaction, leading lower prejudice individuals to treat an outgroup member less positively than they do when they adopt alternative mind-sets; for higher prejudice individuals, perspective taking instead had a positive, albeit less consistent, effect on behavior. The net result was behavior disruption, whereby individuals' treatment of an outgroup member became incongruent with their inner attitudes. This disruption effect was evident for cognitive and affective forms of perspective taking, in ostensible and real face-to-face intergroup interactions, and for feelings of happiness experienced by individuals' interaction partner as well as outside observers' behavior assessments. Results further suggested that self-regulatory effort mediated the effect of perspective taking on intergroup interaction behavior, with the negative consequences of perspective taking for lower prejudice individuals' behavior appearing to stem from complacency rather than trying too hard. Overall, the findings reveal that perspective taking rather than self-focus accounts for the cognitive resource depletion and behavior disruption effects previously demonstrated to stem from evaluative concern in intergroup interaction and indicate that perspective taking may be more reliably helpful outside of intergroup interaction situations than within them. PMID- 19309205 TI - A question of compensation: the social life of the fundamental dimensions of social perception. AB - This research examines the impact of the compensation effect between the fundamental dimensions of warmth and competence on behavioral confirmation. In Experiment 1, participants were presented with 2 groups that varied on 1 of the 2 dimensions and asked to select the questions that they wanted to pose to learn more about the groups. Participants preferred to ask negative (positive) questions about the unmanipulated dimension to the high (low) group. In Experiment 2, participants rated the 2 groups on the basis of naive people answers to those questions. As predicted, compensation emerged. Experiment 3 involved interactions among 3 participants, 1 interviewing the other 2 using the questions selected in Experiment 1. Ratings of targets' reactions again showed compensation. PMID- 19309206 TI - Does contact reduce prejudice or does prejudice reduce contact? A longitudinal test of the contact hypothesis among majority and minority groups in three European countries. AB - A widely researched panacea for reducing intergroup prejudice is the contact hypothesis. However, few longitudinal studies can shed light on the direction of causal processes: from contact to prejudice reduction (contact effects) or from prejudice to contact reduction (prejudice effects). The authors conducted a longitudinal field survey in Germany, Belgium, and England with school students. The sample comprised members of both ethnic minorities (n = 512) and ethnic majorities (n = 1,143). Path analyses yielded both lagged contact effects and prejudice effects: Contact reduced prejudice, but prejudice also reduced contact. Furthermore, contact effects were negligible for minority members. These effects were obtained for 2 indicators of prejudice: negative intergroup emotions and desire for social distance. For both majority and minority members, contact effects on negative emotions were stronger when outgroup contacts were perceived as being typical of their group. Contact effects were also mediated by intergroup anxiety. This mediating mechanism was impaired for minority members because of a weakened effect of anxiety on desire for social distance. Theoretical and practical implications of these findings are discussed. PMID- 19309207 TI - On the malleability of ideology: motivated construals of color blindness. AB - The authors propose that the content of certain sociopolitical ideologies can be shaped by individuals in ways that satisfy their social motivations. This notion was tested in the context of color-blind ideology. Color blindness, when construed as a principle of distributive justice, is an egalitarian stance concerned with reducing discrepancies between groups' outcomes; as a principle of procedural justice, however, color blindness can function as a legitimizing ideology that entrenches existing inequalities. In Study 1, White people high in antiegalitarian sentiment were found to shift their construal of color blindness from a distributive to a procedural principle when exposed to intergroup threat. In Studies 2, 3A, and 3B, the authors used manipulations and a measure of threat to show that antiegalitarian White people endorse color blindness to legitimize the racial status quo. In Study 3B, participants' endorsement of color-blind ideology was mediated by increases in their preference for equal treatment (i.e., procedural justice) as a response to threat. In the Discussion section, the authors examine implications of the present perspective for understanding the manner in which individuals compete over the meaning of crucial ideologies. PMID- 19309208 TI - Motivated response styles: the role of cultural values, regulatory focus, and self-consciousness in socially desirable responding. AB - Three studies investigated the relations between cultural values and socially desirable responding, the processes that underlie them, and factors that influence the strength of the relations. Results indicated that individualism was associated with self-deceptive enhancement but not impression management, whereas collectivism was associated with impression management but not self-deceptive enhancement. Regulatory focus was found to mediate these relations. A promotion focus mediated the relation between individualism and self-deceptive enhancement, whereas a prevention focus mediated the relation between collectivism and impression management. This mediation pattern held regardless of whether individualism and collectivism were determined at the group level (Study 1) or measured at the individual level (Studies 2-3), whether socially desirable responding was operationalized as a scale measure (Studies 1-3) or as reactions to behavioral scenarios (Study 2), and across different measures of regulatory focus. This general mediation pattern was found to be moderated by type of self consciousness (Study 3): The promotion focus mediation was stronger for participants low (vs. high) in private self-consciousness, and the prevention focus mediation was stronger for participants high (vs. low) in public self consciousness. PMID- 19309209 TI - The social costs of emotional suppression: a prospective study of the transition to college. AB - There is growing interest in understanding how emotion regulation affects adaptation. The present study examined expressive suppression (which involves inhibiting the overt expression of emotion) and how it affects a critical domain of adaptation, social functioning. This investigation focused on the transition to college, a time that presents a variety of emotional and social challenges. Analyses focused on 2 components of suppression: a stable component, representing individual differences expressed both before and after the transition, and a dynamic component, representing variance specific to the new college context. Both components of suppression predicted lower social support, less closeness to others, and lower social satisfaction. These findings were robustly corroborated across weekly experience reports, self-reports, and peer reports and are consistent with a theoretical framework that defines emotion regulation as a dynamic process shaped by both stable person factors and environmental demands. PMID- 19309210 TI - Maturation of personality in adolescence. AB - The present research assesses adolescent personality maturation by examining 3 measures of change and stability (i.e., mean-level change, rank-order stability, and profile similarity) of Big Five personality traits, employing data from a 5 annual-wave study with overlapping early to middle (n = 923) and middle to late (n = 390) adolescent cohorts. Results indicated that mean levels of Agreeableness and Emotional Stability increased during adolescence. There was mixed evidence for increases in Extraversion and Openness. Additionally, rank-order stability and profile similarity of adolescent personality traits clearly increased from early to late adolescence. For all change facets, the authors found evidence for gender differences in the timing of adolescent personality maturation, as girls were found to mature earlier than boys. PMID- 19309211 TI - Dispositional optimism and engagement: the moderating influence of goal prioritization. AB - Research indicates that a positive relationship generally exists between dispositional optimism and goal engagement and attainment. The authors argue, however, that dispositional optimism may not always be associated with more active goal pursuit. Rather, they hypothesized that this relationship is moderated by how highly a goal is prioritized. For high-priority goals, they predicted that optimistic individuals would indeed increase goal engagement and would be more likely to attain their goal relative to individuals low in optimism. For low-priority goals, they anticipated that optimistic individuals would not display greater goal engagement or attainment. In 5 studies they assessed these predictions across a variety of domains, including friendship formation, exercise persistence, and scholastic achievement. Results supported their contention that goal priority acts as a moderator of the relationship between dispositional optimism and both goal engagement and goal attainment. Evidence of 1 mediator of this moderation effect-behavioral intentions-and of a limiting factor-the temporal ordering of goals-is also presented. PMID- 19309212 TI - Memorial consequences of answering SAT II questions. AB - Many thousands of students take standardized tests every year. In the current research, we asked whether answering standardized test questions affects students' later test performance. Prior research has shown both positive and negative effects of multiple-choice testing on later tests, with negative effects arising from students selecting incorrect alternatives on multiple-choice tests and then believing they were correct (Roediger & Marsh, 2005). In the current experiments, undergraduates and high school students answered multiple-choice questions retired from SAT II tests (that are no longer in the testing pool) on biology, chemistry, U.S. history, and world history, and later answered cued recall questions about these subjects. In 3 experiments, we observed positive testing effects: More final cued-recall questions were answered correctly if the items had appeared on the initial multiple-choice test. We also sometimes observed negative testing effects: intrusions of multiple-choice distractors as answers on the final cued-recall test. Students who scored well on the initial test benefited from taking the test, but lower achieving students showed either less benefit (undergraduates) or costs from the testing (high school students). PMID- 19309214 TI - A perceptual process approach to selecting color scales for complex visualizations. AB - Previous research has shown that multicolored scales are superior to ordered brightness scales for supporting identification tasks on complex visualizations (categorization, absolute numeric value judgments, etc.), whereas ordered brightness scales are superior for relative comparison tasks (greater/less). We examined the processes by which such tasks are performed. By studying eye movements and by comparing performance on scales of different sizes, we argued that (a) people perform identification tasks by conducting a serial visual search of the legend, whose speed is sensitive to the number of scale colors and the discriminability of the colors; and (b) people perform relative comparison tasks using different processes for multicolored versus brightness scales. With multicolored scales, they perform a parallel search of the legend, whose speed is relatively insensitive to the size of the scale, whereas with brightness scales, people usually directly compare the target colors in the visualization, while making little reference to the legend. Performance of comparisons was relatively robust against increases in scale size, whereas performance of identifications deteriorated markedly, especially with brightness scales, once scale sizes reached 10 colors or more. PMID- 19309213 TI - Experience-based mitigation of age-related performance declines: evidence from air traffic control. AB - Previous research has found age-related deficits in a variety of cognitive processes. However, some studies have demonstrated age-related sparing on tasks where individuals have substantial experience, often attained over many decades. Here, the authors examined whether decades of experience in a fast-paced demanding profession, air traffic control (ATC), would enable older controllers to perform at high levels of proficiency. The authors also investigated whether older controllers would show diminished age-related decrements on domain-relevant cognitive abilities. Both young and old controllers and non-controllers performed a battery of cognitive and ATC tasks. Results indicate that although high levels of experience can reduce the magnitude of age-related decline on the component processes that underlie complex task performance, this sparing is limited in scope. More important, however, the authors observed experience-based sparing on simulated ATC tasks, with the sparing being most evident on the more complex air traffic control tasks. These results suggest that given substantial experience, older adults may be quite capable of performing at high levels of proficiency on fast-paced demanding real-world tasks. The implications of these findings for global skilled labor shortages are discussed. PMID- 19309215 TI - Humor in advertisements enhances product liking by mere association. AB - Humor in advertising is known to enhance product liking, but this attitude change is often considered nonpredictive of product choice. Previous research relied exclusively on explicit self-report measures to assess attitudes and purchase intentions. The present research shows that unobtrusive association of a product with humor can affect persuasion through implicit attitude change. Participants viewed humorous and nonhumorous cartoons in a mock-up magazine. One of two products was consistently presented in the vicinity of the humorous cartoons, whereas the other product was consistently presented in the vicinity of the nonhumorous cartoons. The results of an evaluative priming task showed enhanced evaluations of products paired with humor (Experiment 1, 2, and 3). Furthermore, these enhanced evaluations mediated the relation between association with humor and product choice (Experiment 2 and 3). Paradoxically, products paired with humor were also less recognized than the control products (Experiments 2 and 3). In summary, the present research demonstrates that mere association with humor enhances product evaluations and product choice in a way that is dissociated from the accessibility of the product in memory. PMID- 19309216 TI - Fast and confident: postdicting eyewitness identification accuracy in a field study. AB - The combined postdictive value of postdecision confidence, decision time, and Remember-Know-Familiar (RKF) judgments as markers of identification accuracy was evaluated with 10 targets and 720 participants. In a pedestrian area, passers-by were asked for directions. Identifications were made from target-absent or target present lineups. Fast (optimum time boundary at 6 seconds) and confident (optimum confidence boundary at 90%) witnesses were highly accurate, slow and nonconfident witnesses highly inaccurate. Although this combination of postdictors was clearly superior to using either postdictor by itself these combinations refer only to a subsample of choosers. Know answers were associated with higher identification performance than Familiar answers, with no difference between Remember and Know answers. The results of participants' post hoc decision time estimates paralleled those with measured decision times. To explore decision strategies of nonchoosers, three subgroups were formed according to their reasons given for rejecting the lineup. Nonchoosers indicating that the target had simply been absent made faster and more confident decisions than nonchoosers stating lack of confidence or lack of memory. There were no significant differences with regard to identification performance across nonchooser groups. PMID- 19309217 TI - Lineup administrator influences on eyewitness identification decisions. AB - The present research examines how a lineup administrator may influence eyewitness identification decisions through different forms of influence, after providing the witness with standard, unbiased instructions. Participant-witnesses viewed a staged crime and were later shown a target-present or target-absent lineup. The lineup administrators either remained silent while the witness examined the lineup, made ostensibly cautionary statements to the witness, or prompted the witness to identify the person in the lineup who seemed most similar to the perpetrator. These two forms of influence, denoted as subtle-influence and similarity-influence conditions, led to different patterns of identification results. Results for the similarity-influence condition were generally consistent with criterion shift and relative judgment models of eyewitness decision making. Results for the subtle-influence condition, however, cannot be explained by alterations in the decision rule. A weighted matching model is outlined to explain results from the subtle-influence condition. Witnesses seemed generally unaware of the attempts by the lineup administrator to influence their decision, although some noted it, and the probative value of suspect identifications was lower for those who did note it. Implications for theory and policy are discussed. PMID- 19309218 TI - Exploring the diagnostic utility of facial composites: beliefs of guilt can bias perceived similarity between composite and suspect. AB - Facial composite research has generally focused on the investigative utility of composites-using composites to find suspects. However, almost no work has examined the diagnostic utility of facial composites-the extent to which composites can be used as evidence against a suspect. For example, detectives and jurors may use the perceived similarity of a suspect to a composite as evidence to determine the likelihood of a suspect's guilt. However, research in social cognition and models of cognitive coherence suggest that these similarity judgments may be biased by evaluators' preexisting beliefs of guilt. Two studies examined how preexisting beliefs of guilt influence similarity ratings between a suspect and a facial composite. Study 1 (n = 93) demonstrated that mock investigators' beliefs in a suspect's guilt inflated their subsequent similarity ratings. Study 2 (n = 49) demonstrated that mock-jurors' beliefs in a defendant's guilt predicted their similarity ratings. These findings highlight a problem of using facial composites as evidence against a suspect, and demonstrate the malleability of similarity judgments. PMID- 19309219 TI - Cytarabine conjugates with biologically active molecules and their potential anticancer activity. AB - The presented review article deals with various conjugates of arabinosylcytosine (araC). This powerful drug that is routinely used in therapy of hematological malignancies has some shortcomings, which limit its use and therapeutic effects. These are low lipophilicity, low stability to degrading enzymes and need for biological activation through phosphorylation. Conjugating araC to another molecule is done with the intention of increasing araC stability and lipophilicity and possibly avoiding rate-limiting araC phosphorylation. An attachment of that another molecule, possessing its own biological activity, may result in formation of a conjugated molecule with new biological activities and better therapeutic potential. The review deals with various araC conjugates formed at the positions N(4), 2, 2', 3' and 5'. Biological activities and differences from araC of compounds formed by conjugation are also discussed. PMID- 19309220 TI - Cancer and depression: a prospective study. AB - Cancer diagnosis and treatment often produce psychologic stresses resulting from the actual symptoms of the disease, as well as from perceptions of the disease and its stigma. Depression is seen in many cancer patients. Depression occurs in approximately 25% of palliative care patients. It is widely recognised by clinicians that depression is a difficult symptom to identify amongst patients with advanced illness. The study is aimed for screening of depression among palliative care female patients. This study was local, prospective and cross sectional. It was carried at Department of Clinical Oncology and Radiation Therapy of Charles University Hospital in Hradec Kralove, Czech Republic. Dates were obtained during year 2007 - 2008 in 64 palliative care female patients. The mean age for all 64 subjects was 60,5 years old (aged 29 - 88 years old). The Czech version of Zung self-rating depression scale was performed. The statistical evaluation presents that mean SDS (self-rating depression score) certifies the presence of signs of mildly depression among palliative care female patients (SDS range was 50-59). The mean SDS in all subjects was 56. The mean SDS in group of healthy females was 38,9 (normal range). The incidence of depression is 71,8% (46 of all 64 subjects). The relevance of depression is characterized: severely depressed was proved in 8 of all 46 subjects, the moderately depressed in 21 subjects of all 46 subjects and mildly depressed in 17 of all 46 subjects. The statistical evaluation not presents statistically significant dependence of SDS on smoking abuse, marital status, age, number of associated diseases and type of palliative care. The statistical evaluation presents that patients with cancer of lung, with cancer of endometrium, with cancer of gallbladder and with melanomas are moderately depressed (SDS 60-69), patients with cancer of ovary, with cancer of breast, with primary brain tumour, with cancer of ventricle, with cancer of pancreas head and with cancer of bucall cavity are mildly depressed (SDS 50-59). The results show that subsists clear association between oncological disease in palliative care and depression. PMID- 19309221 TI - Isolation and properties of gene-modified mouse bcr-abl-transformed cells expressing various immunostimulatory factors. AB - B210 cells are murine (BALB/c) cells transformed by bcr-abl fusion gene. After intravenous administration they are capable of inducing leukaemia-like disease in syngeneic mice. From these cells a thymidine-kinase less subline was derived. It was significantly less pathogenic than the parental cells. However, a highly pathogenic clone denoted B210cTK-/cl-2 was isolated from its population. As determined by Western blotting, these cells produced more p210 protein than the parental B210 cells. To successfully transfect these cells a modified electroporation method was introduced. Bicistronic plasmids carrying gene for herpes simplex thymidine kinase (HSV TK) and the gene for either granulocyte monocyte colony stimulation factor (GM-CSF), interleukin-2 (IL-2) or interleukin 12 (IL-12) were used for the transfection experiments. Gradually, cell lines producing these cytokines were isolated in media supplemented with hypoxantin, aminopterin and thymidine (HAT). All of them were highly sensitive to ganciclovir in vitro confirming that the cells produced HSV TK. The genetic modification of B210cTK-/cl-2 was associated neither with the alteration of p210 bcr-abl production nor with any changes in expression of MHC class I molecules. From populations of each of the three lines several cell clones were isolated and tested for the production of the respective cytokines. The original uncloned population and several clones differing in the cytokine production were administered intravenously into mice. All animals survived without symptoms of the disease suggesting that the gene-modification was associated with the loss of pathogenicity. KEYWORDS: CML, Bcr-Abl, HSV TK, cytokines, gene-modified tumour cells, pathogenicity. PMID- 19309222 TI - Imatinib is a substrate for various multidrug resistance proteins. AB - An increasing resistance to imatinib is an emerging problem in patients with chronic myeloid leukemia (CML). The aim of the study was to asses mechanisms related to cellular drug resistance in imatinib-resistant derivates of chronic myeloid leukemia K-562 cell line. A parental K-562 and its imatinib-resistant derivate cell lines were used. Cell lines were tested for cytotoxicity of imatinib, cytarabine, busulfan and etoposide by the MTT assay. The cytotoxicity was expressed as IC50, inhibitory concentration for 50% of cells. Multidrug resistance proteins expression, rhodamine retention and daunorubicin accumulation were measured for each cell line. Continuous exposition of K-562 cell line to 0.01-0.02 mM imatinib resulted in development of resistance, while exposition to 0.1 microM imatinib increased cell sensitivity to this drug. There was a high correlation between PGP, MRP1 and LRP expression and IC50 values for imatinib and etoposide. All tested cell lines were highly resistant to cytarabine. Rhodamine retention alone and in the presence of cyclosporine was the lowest in imatinib resistant K-562R-0.1 cell line, what suggest high PGP activity in this cell line. The highest daunorubicin accumulation was observed in parental K-562 cell line, while it was lower in imatinib-resistant cell lines. These data suggest that imatinib is a substrate for multidrug resistance proteins, and an increased expression of PGP, MRP1 and LRP play a role in resistance to imatinib in CML. KEYWORDS: imatinib, multidrug resistance proteins, chronic myeloid leukemia, PGP, MRP1, LRP. PMID- 19309223 TI - Administration of isothiocyanate (E-4IB) and cisplatin leads to altered signalling and lysosomal export in human ovarian carcinoma sensitive- and cisplatin-resistant cells. AB - The aim of this study was to compare the effect of a new synthetic isothiocyanate derivative, ethyl 4-isothiocyanatobutanoate (E-4IB) and cisplatin (CDDP) in CDDP sensitive human ovarian carcinoma cell line (A2780) and its resistant subline (A2780/CP). In parental cells, in comparison to untreated cells, sequential administration of both compounds led to higher exosomal dye (LysoTracker Green DND-26) retention and to alterations of mitogen-activated protein kinases (MAPKs), JNK, ERK and p38, or Akt kinase accompanied by changes in several anti- and pro-apoptotic molecules and lysosomal protein LAMP-1, as detected by Western blotting. On the contrary, variant A2780/CP cells were resistant to CDDP- or to combined sensitizer (E-4IB)/inducer (CDDP)-related apoptosis induction and exerted minor changes in the levels of these molecules. PMID- 19309224 TI - Prognostic factors in patients with relapsed or primary refractory germ cell tumors. AB - The aim of the study was to define prognostic factors of overall and event- free survival in patients with germ cell tumors progressing after platinum-based induction chemotherapy with or without surgery. A total of 98 progressing patients were identified out of 700 patients with germ cell tumors treated with platinum-based induction chemotherapy in National Cancer Institute in Bratislava with or without surgery. 98 progressing patients received first salvage chemotherapy from October 1986 to November 2007 due to progression after a previous partial or complete response to induction chemotherapy as well as patients who failed to achieve favourable response to primary therapy. Prognostic factors of survival and event-free survival after first salvage chemotherapy were assessed by univariate analysis. For all 98 progressing patients the median time from the start of induction chemotherapy to progression was 10,2 months (range: 0 256,7 months). 24 (24 %) patients relapsed after 2 years. Median overall survival time following progression was 25,4 months. Estimated 2- and 5- year overall survival rate for all progressing patients was 46 % (95 % CI 41-61%) and 24 % (95% CI 31-51%) respectively. Survival after first salvage chemotherapy was significantly enhanced for patients with age more than 40 years at primary diagnosis, nonvisceral metastasis at the time of induction chemotherapy, prior CR to induction chemotherapy, progression-free interval > 2 years, serum human chorionic gonadotropin level at relapse above or bellow 100 IU/l, a normal serum lactate dehydrogenase level at relapse, one site of metastasis at relapse, treatment with cisplatin-based first salvage chemotherapy, first regimen VIP and favourable response to salvage chemotherapy. Estimated 2- and 5-year event-free survival rate for all patients was 30% (95% CI 24-43% ) and 16%(95% CI 19-37% ) respectively. As a significant favourable prognostic factors of event-free survival were identified: prior CR to induction chemotherapy, progression-free interval > 2 years, one site of metastasis at relapse, treatment with cisplatin based first salvage chemotherapy, first line salvage regimen VIP and favourable response to salvage chemotherapy. Identification of prognostic features in patients with germ cell tumors progressing after platinum-based induction chemotherapy may direct salvage therapy and requires further investigation of new combination of salvage therapy for those with poor prognosis. Our study showed the indispensable revaluating of chemosenzitivity in patients with late relapses and therapeutic value of additive surgical approach after salvage chemotherapy in patients with reccurent germ cell tumors. PMID- 19309225 TI - Inhibition of cell proliferation and arrest of cell cycle progression by blocking chloride channels in human laryngeal cancer cell line Hep-2. AB - Chloride channel (ClC) is involved in normal physiological processes and pathology of various diseases. Although it is recognized that blockade of ClC inhibits the cell proliferation, it is not well understood the potential function of ClC in laryngeal cancer. In this study, we investigated the effect of the ClC inhibitor on cell proliferation, cell cycle progression in human laryngeal cancer cell line Hep-2, as well as the effect on the phosphorylation levels of ERK1/2 and AKT1. In this study crystal violet method was used to study the effect of the ClC inhibitor, 5-nitro-2-(3-phenylpropylamino) benzoic acid, NPPB, on Hep-2 cell proliferation. The impaction of the inhibitor on the cell cycle distribution was investigated by the flow cytometry (FCM). Western blot was performed to measure the phosphorylation levels of ERK1/2 and AKT1. Our data indicated ClC played an important role in Hep-2 cell proliferation and cell cycle. NPPB inhibited Hep-2 cell proliferation when compared with the controls. Blockade of ClC arrested cell cycle progression and suppressed the phosphorylation of ERK1/2 and AKT1 in Hep-2 cells by inhibition of cell proliferation by CIC inhibitor (NPPB) could be through arresting cell cycle progression, which is probably by suppressing phosphorylation of ERK1/2 and AKT1. PMID- 19309226 TI - The detection of circulating tumor cells expressing E6/E7 HR-HPV oncogenes in peripheral blood in cervical cancer patients after radical hysterectomy. AB - The aim of this study was to establish the sensitive, specific and clinically acceptable method for detection of tumor cells (TCs) circulating in peripheral blood (PB) of cervical cancer patients without the clinically detectable risk of disease progression. The 7.5 ml of PB of healthy donor was spiked with 5 to 100 cells from SiHa or HeLa cell lines. The spiked tumor cells were collected without gradient centrifugation, by standard gradient centrifugation or by modified gradient centrifugation combined with immunomagnetic separation using EpCAM antibody with affinity for epithelial cell adhesion molecule. The number of collected TCs was determined by EpCAM-FITC-staining and their viability was detected by nested RT-PCR amplifying E6/E7 HR-HPV 16 or HR-HPV 18 oncogenes. For the technical validation of this approach the TCs separation and RT-PCRs were repeated several times. The recovery of viable TCs was reproducibly higher using modified gradient centrifugation combined with immunomagnetic separation in comparison with standard approach. The recovery of TCs in low number of spiked TCs (range from 5 - 20 TCs in 7.5 ml of PB) using modified gradient centrifugation was not reproducible. The recovery of TCs in higher number of spiked TCs (25 TCs and more in 7.5 ml of PB) was reproducible with average recovery about 50 %. The sensitivity of nested RT-PCR amplifying E6/E7 oncogenes was decisively influenced by the number of recovered TCs and the amount of cDNA introduced to RT-PCR, as well. Using this approach we were allowed to detect circulating TCs (CTCs) in cervical cancer patients without metastases, thus this procedure might become a tool to early estimation of disease progression. According to our knowledge, this is the first report describing the use of EpCAM antibody for CTCs detection in cervical cancer patients. PMID- 19309227 TI - Expression of p53, cyclin D1 and EGFR correlates with histological grade of adult soft tissue sarcomas: a study on tissue microarrays. AB - Soft tissue sarcomas (STSs) are rare heterogeneous tumors with variable clinical course and outcome. The management of STSs depends upon the accurate histopathological diagnosis and assessing their histological grade. Currently, core needle biopsies are becoming increasingly popular for diagnosing STSs but value of histological grading is limited from this type of specimens. To evaluate the immunohistochemical expression of p53, mdm-2, cyclin D1, p16, nm23, EGFR and Ki-67 labelling index in adult STSs patients and their association with histological grade of STSs, we analysed 101 primary untreated STSs of the limbs and trunk using the tissue microarray technique on formalin-fixed, paraffin embedded tissue samples. The cases consisted of 15 G1, 28 G2 and 58 G2 sarcomas. Ki-67 labelling index (LI) was calculated from whole block sections for the possibility to select the most proliferative regions. The LI ranged from 1.26 to 75.5% (median 26.7%) and strongly correlated with the mitotic count (p rs for adult patients with STSs and may assist in establishment of the histological grade in STSs. PMID- 19309228 TI - Epithelial bone marrow cells in patients with advanced esophageal squamous cell carcinoma. AB - The aim of the current study was to examine epithelial cells in the bone marrow and peripheral blood of patients with various stages of esophageal squamous cell cancer prior to surgical treatment and to analyze the prognostic significance of these carcinoma cells deposits to the stage of the disease and applied surgical therapy. Thirty-two patients (25 men and 7 women), and 5 healthy bone marrow donors serving as controls were studied. Bone marrow samples were evaluated by light microscopy and examined by flow cytofluorometry. Cells were phenotypically analyzed for the antigens CD45- and CD18+ and/or EMA+. Results are presented as the number of cells revealing the investigated phenotype per 10 (5)analyzed cells. CD18 was expressed in the bone marrow cells of 15 of the 32 (47%) patients and EMA in 20/32 (62%), but not in peripheral blood. In 13 of the 32 pts (41%), co-expression of CD18 and EMA was observed. Patients with the proportion of marrow erythroblasts below 15% had higher numbers of CD18+ and EMA+ cells and there was a negative correlation between the number of erythroblasts and EMA+ cells (r=0.54, p=0.01). In patients with esophageal cancer and anemia, the number of EMA+ cells was higher (p=0.05) and the percentage of erythropoietic cells in the bone marrow was lower (p=0.01). In conclusion, flow cytofluorometry using anti-cytokeratin and anti-EMA antibodies may be useful in evaluating microdeposits of esophageal squamous cells in bone marrow. A dysfunctioning erythropoietic system causing anemia can be a first signal for the presence of malignant cell microdeposits in the marrow of patients with esophageal carcinoma. PMID- 19309229 TI - Celecoxib and melatonin in prevention of female rat mammary carcinogenesis. AB - The present experiment aims to evaluate tumor suppressive effects of a selective inhibitor of cyclooxygenase-2 (COX-2) celecoxib (Celebrex, Pfizer) administered alone and in combination with melatonin in the prevention of N-methyl-N nitrosourea (NMU)-induced mammary carcinogenesis in Sprague-Dawley female rats. Celecoxib was administered daily at a concentration of 1.666 g/kg diet to two groups during 20 weeks (starting a week before first NMU application). A combination of celecoxib and melatonin applied in drinking water (20 microg/ml drinking water), daily from 15:00 to 08:00 hours was administered to the second group. The anticarcinogenic effects of chemopreventive drugs were compared with control (NMU) animals. Celecoxib administration decreased mammary tumor incidence (by 24%), while combination of celecoxib and melatonin decreased tumor incidence even more significantly (-30%). Significant decrease in tumor frequency per group was recorded in both groups with chemoprevention: celecoxib alone (-54%) and combination of celecoxib and melatonin (-64%). Celecoxib significantly influenced tumor frequency per animal in the group with combination of both protective substances (-52%). Celecoxib administration resulted in prolonged latency by 3%, and by 13% in the group with combination of both protective substances. These results confirm preventive effects of celecoxib in induced rat mammary carcinogenesis. The administration of isolated MEL had only lesser effect, but in the combination with CELE revealed some potentiating influence in mammary carcinogenesis inhibition. The present study is the first to prove efficacy of the above-mentioned celecoxib and melatonin intake. Our results point to the need for a deeper analysis of coxib efficacy in human carcinogenesis. PMID- 19309230 TI - Autologous Hsp70 immunization induces anti-tumor immunity and increases longevity and survival of tumor-bearing mice. AB - Heat Shock proteins 70 (Hsp70) is a family of highly conserved molecules that maintain the function of crucial cellular pathways during stress. Hsp70 derived from tumor cells is bound with tumor antigenic peptides from the diverse antigen cytosolic pool. Tumor-derived Hsp70 preparations after ex vivo administration permit antigen presenting cells (APCs) to present tumor antigen to their cell surface and induce tumor specific immunity in many types of malignancies by directly eliciting cytotoxic T lymphocytes (CTL) response. In the present investigation, we have demonstrated that immunization with tumor cell derived Hsp70 lead to an effective survival advantage in mice with minimal residual tumor cells from which Hsp70 is derived, by involvement of immune cell types in the rejection of tumor in the Hsp70 immunized tumor-bearing host mice and their post immunization cytokine repertoire. It has been observed that autologous Hsp70 induces specific anti-tumor immunity and effectively eradicates tumors in the host mice, thereby enhancing survival of tumor-bearing host. Immunization with tumor-derived autologous Hsp70 effectively primed specific CTL response and increased tumor cell lysis independently of CD4 (+)T lymphocytes. Increase in type I cytokines in the serum of Hsp70 immunized mice was also observed that indicates its adjuvant property in the host. Furthermore, Hsp70 immunized mice did not show any systemic disorder. Therefore, it could be assumed as safe and might be clinically useful for vaccination against malignant human tumors. PMID- 19309231 TI - Metformin in chemically-induced mammary carcinogenesis in rats. AB - In this paper the chemopreventive effect of peroral antidiabetic metformin in mammary carcinogenesis in female Sprague-Dawley rats was evaluated. Mammary carcinogenesis was induced by N-methyl-N-nitrosourea (NMU) administered in two intraperitoneal doses each per 50 mg/kg b.w. between 43.-55. postnatal days. Metformin was administered in drinking water (at a concentration of 50 microg/ml and 500 microg/ml) 13 days before the first NMU dose until the termination of the experiment. During the experiment the animals were weekly weighed and palpated for the presence of mammary tumors, the incidence, latency, tumor frequency, and tumor volume were recorded. The experiment was terminated 18 weeks after the first NMU dose, basic tumor growth parameters and metabolic and hormonal variables were evaluated. Metformin did not significantly alter the tumor growth although a delay in tumor onset was recorded after higher metformin dose. Metformin altered metabolic and hormonal variables. Insulinemia decreased after both metformin doses in comparison with intact rats without changes in glycemia, triacylglycerols concentration was decreased in liver and increased in serum when compared to intacts. Higher metformin dose attenuated lipoperoxidation in liver. PMID- 19309232 TI - Genetic analysis of KRAS mutation status in metastatic colorectal cancer patients. AB - Colorectal carcinoma (CRC) represents a serious problem worldwide: in the Slovak republic are diagnosed about 2600 new CRC cases annually and its incidence is increasing. Colorectal cancer patients may succumb to the disease because of local recurrence or local formation of metastasis. Therefore, it is necessary to modulate therapeutic algorithm with new methods, leading to early diagnostic of CRC or changing the existing therapeutic procedures. Recent progresses have been made in understanding of EGFR pathway involved in CRC carcinogenesis, especially the role of Ras protein. Mutations in KRAS oncogene are frequently found in human cancers, particularly colorectal, pancreatic, billiary tract and lung tumors. The presence of the KRAS mutations in metastatic colorectal cancer patients correlates with lack of response to the certain epidemal growth factor receptor (EGFR) inhibitor therapies, such as Panitumumab and Cetuximab. Consequently, screening for KRAS mutations status may be used as a prognostic marker, because the CRC patients with KRAS positive tumors have a worse prognosis. The aim of our study was to establish the methods for rapid and sensitive detection of KRAS mutation status in formalin fixed paraffin embedded (FFPE) tissues DNA. We applied Real Time PCR analysis (TheraScreen KRAS Mutation Test Kit) and sequencing analysis (optimised for the analysis of FFPE tissues) to detect somatic mutations in codon 12 and 13 of KRAS gene. Both methods were used concurrently in the panel of DNA isolated from 25 colorectal FFPE tissues tumor. The positive or negative results from all 25 samples were identified by both methods independently. The KRAS mutations were presented in 8 of 25 patients (32%). Our results demonstrate that the Real Time PCR analysis can be used for detection of somatic KRAS mutations in FFPE clinical samples. However, we also recognize that the sequencing analysis of approximately 200bp amplicons may be used for mutations status screening, but with care of method sensitivity. PMID- 19309233 TI - Brain response to induced peripheral cancer development in rats: dual fos tyrosine hydroxylase and fos-oxytocin immunohistochemistry. AB - OBJECTIVE: During last few decades a considerable number of data has emerged supporting the hypothesis that central nervous system might monitor and modulate tumor growth. This assumption is based on two facts: 1. immune system plays a crucial role in the development and progression of cancer; 2. immune and nervous systems communicate tightly and bidirectionally. The aim of present study was to elucidate whether tumor growth may induce detectable changes in brain structures that are involved in the response to immune challenges. METHODS: Using Fos immunohistochemistry, we investigated whether the advanced stage of cancer, induced by a single intraperitoneal injection of BP6-TU2 fibrosarcoma cells to male Wistar rats, could activate Fos expression in the nucleus of the solitary tract (NTS), amygdala and parabrachial nuclei (PBN) and also activate some of neuronal phenotypes including tyrosine hydroxylase (TH) neurons in the brainstem noradrenergic cell groups and hypothalamic oxytocinergic neurons. RESULTS: Twenty eight days after the initiation of tumor process we found increased Fos expression in NTS/A2, A1 noradrenergic cells, PBN as well as in the hypothalamic paraventricular, supraoptic and accessory oxytocinergic neurons. These structures are involved in the transmission of signals related to immune challenges within the brain and consequent elaboration of neuro-endocrine responses. CONCLUSIONS: The data obtained are supporting the view that the information on peripheral tumor development might be transmitted to the brain. However, further studies are necessary to be performed to reveal whether our findings can be attributed to specific effect of cancer or whether observed changes in the activity of brainstem and hypothalamic neurons reflex processes that only accompany the cancer progression. PMID- 19309234 TI - Cannabinoid-mediated regulation of the hypothalamo-pituitary-adrenal axis in rats: age dependent role of vasopressin. AB - OBJECTIVE: Adaptation to stress is a fundamental component of life and the hypothalamo-pituitary-adrenocortical axis (HPA) plays a crucial role in it. The place of cannabinoid influence seems to be in the brain, especially where corticotropin releasing hormone and vasopressin (AVP) secreting neurons are located. The role of AVP is considered to be more important in young than in adult rats. Here we addressed the question if cannabinoid-mediated regulation of the HPA involves AVP and if there is any difference between young and adult rats in this process. METHODS: 10-day-old and adult AVP deficient Brattleboro rats were compared with their heterozygous littermates 1h after WIN 55,212-2 (6mg/kg i.p.) injection. RESULTS: In control animals the injection led to elevated adrenocorticotropin (ACTH) and corticosterone hormone levels at both ages without remarkable age difference in ACTH levels while all corticosterone levels of adults was approximately 10-times higher. The ACTH secretion of young AVP deficient rats failed to react to WIN 55,212-2 injection while their corticosterone levels were even higher than their littermates. In contrast in adult the role of AVP was diminished. CONCLUSIONS: We can conclude that the peripheral administration of cannabinoids leads to HPA axis stimulation, which process involves AVP at least in the young rats. The discrepancy between ACTH and corticosterone levels in young rats suggests an alternative adrenal gland regulatory pathway, which might be present in all studied animals. However, it comes to the front just in AVP deficient pups. PMID- 19309235 TI - Opioid antagonist-induced regulation of the mu-opioid receptor expression in MCF 7 breast cancer cell line. AB - OBJECTIVE: In our earlier study we have demonstrated that MCF-7 cell line expresses all three opioid receptor types, micro, delta and kappa (MOR, DOR and KOR, respectively), but predominantly MOR. Morphine, as well as endogenous MOR selective agonists, endomorphin-1 and endomorphin-2 were shown to decrease MOR gene expression in MCF-7 cells. Opioid antagonist - naloxone -produced the opposite effect, increasing MOR gene expression. In this study we investigated and compared the influence of several opioid antagonists of alkaloid structure (beta-funaltrexamine and naloxonazine) and of peptide structure: [Dmt(1), D-1 Nal(4)]endomorphin-2, [Dmt, D-2-Nal(4)]endomorphin-1, and [Dmt, D-2 Nal(4)]endomorphin-2 on MOR up-/down-regulation and proliferation in MCF-7 cell line. METHODS: MCF-7 cells were incubated with opioids. The levels of MOR mRNA were assessed using quantitative real-time RT-PCR assay. Cell growth was measured by Mosmann tetrazolium salt assay. RESULTS: It was shown that all tested opioid antagonists produced up-regulation of MOR gene expression, but the strongest effect was observed with naloxonazine. However, none of the antagonists at concentrations as high as 10(-4) M showed any antiproliferative effects on MCF-7 cells, neither in the presence or absence of beta-estradiol. CONCLUSION: It seems that up- or down-regulation of MOR mRNA levels has no direct effect on cell proliferation. PMID- 19309236 TI - Reference values of body composition for adult females who are classified as normal weight, overweight or obese according to body mass index. AB - OBJECTIVE: The aim of this study was to establish reference values for body composition as measured by Bioelectrical Impedance Analyser (BIA) in adult females without any endocrinological and/or metabolic disorders, according to their body mass index (BMI) grouped as normal, overweight, obese and morbid obese. PATIENTS AND METHODS: This study was performed in a total of 327 adult females. In addition to the estimation of their body weight, height, waist and hip circumference, their body composition was measured by BIA. The results were statistically evaluated by a computer program using ANOVA test. RESULTS: Together with the increase of BMI also the percentage of body fat and basal metabolism (BM) increased significantly, while the percentage of body water, fat free mass as well as lean/fat ratio showed a significant decrease. In addition, waist and hip ratio, percentage of body fat and BM showed a significant positive corelation with BMI. CONCLUSION: These results are recommended to be used as reference values for the studies on body composition, especially to predict the degree of body fatness of obese patients and also nutritional status of patients who need nutritional supports. PMID- 19309237 TI - Hormone-dependent and hormone-independent control of metabolic and developmental functions of malate dehydrogenase - review. AB - Malate dehydrogenases (MDHs) are specific class of ubiquitous and multimeric oxidative decarboxylases with well conserved amino acid sequences in structurally important regions and with similar overall structural topology. They mostly use malate or oxaloacetate as substrates to generate pyruvate and utilize preferentially NADP or NAD as cofactor. Among species and even within an organism they differ in their subcellular localization and specificity for the cofactor. Comparison across microbial, plant and animal kingdoms show that MDHs were able to adopt tissue-, species- or environmental-specific functions while still keeping main structural features. Although basic principles of MDH regulation are similar to other enzymes and include oligomerization, cofactor binding, divalent cation availability, some of MDH enzymes are regulated also at different levels involving control of hysteresis, protein-protein interaction and gene expression. In this review we concentrate on those aspects of MDH function and regulation in animals that are specifically associated with cell differentiation and proliferation, ontogenic development, hormonal control, and partly with diseases. Accenting these aspects of MDHs provides emerging and new views on their regulatory function in complex eukaryotic metazoan organisms that goes beyond their classical role in basic metabolism. PMID- 19309238 TI - Vascularization--the conduit to viable engineered tissues. AB - Long-term viability of thick three-dimensional engineered tissue constructs is a major challenge. Addressing it requires development of vessel-like network that will allow the survival of the construct in vitro and its integration in vivo owing to improved vascularization after implantation. Resulting from work of various research groups, several approaches were developed aiming engineered tissue vascularization: (1) embodiment of angiogenesis growth factors in the polymeric scaffolds for prolonged release, (2) coculture of endothelial cells with target tissue cells and angiogenesis signaling cells, (3) use of microfabrication methods for creating designed channels for allowing nutrients to flow and/or for directing endothelial cells attachment, and (4) decellularization of organs and blood vessels for creating extracellular matrix. A synergistic effect is expected by combining several of these approaches as already demonstrated in some of the latest studies. Current paper reviews the progress in each approach and recent achievements toward vascularization of engineered tissues. PMID- 19309239 TI - Modulation of 3D fibrin matrix stiffness by intrinsic fibrinogen-thrombin compositions and by extrinsic cellular activity. AB - Fibrin is a substance formed through catalytic conversion of coagulation constituents: fibrinogen and thrombin. The kinetics of the two constituents determines the structural properties of the fibrin architecture. We have shown previously that changing the fibrinogen and thrombin concentrations in the final three-dimensional (3D) fibrin matrix influenced cell proliferation and differentiation. In this study, we further examined the effect of changing fibrinogen and thrombin concentrations in the absence or presence of fibroblasts on the structural modulus or stiffness of 3D fibrin matrices. We have prepared fibroblast-free and fibroblast-embedded 3D fibrin matrices of different fibrinogen and thrombin formulations, and tested the stiffness of these constructs using standard mechanical testing assays. Results showed that there was a corresponding increase in stiffness with increasing thrombin and fibrinogen concentrations; the increase was more notable with fibrinogen and to a lesser degree with thrombin. The effect of fibroblasts on the stiffness of the fibrin construct was also examined. We have observed a small increase in the stiffness of the fibroblast-incorporated fibrin construct as they proliferated and exhibited spreading morphology. To our knowledge, this is the first comprehensive report detailing the relationship between fibrinogen and thrombin concentrations, cell proliferation, and stiffness in 3D fibrin matrices. The data obtained may lead to optimally design suitable bioscaffolds where we can control both cell proliferation and structural integrity for a variety of tissue engineering applications. PMID- 19309240 TI - Composite chitosan/nano-hydroxyapatite scaffolds induce osteocalcin production by osteoblasts in vitro and support bone formation in vivo. AB - There is a significant clinical need to develop alternatives to autografts and allografts for bone grafting procedures. Porous, biodegradable scaffolds based on the biopolymer chitosan have been investigated as bone graft substitutes, and the addition of calcium phosphate to these scaffolds has been shown to improve the mechanical properties of the scaffold and may increase osteoconductivity. In this study, in vitro mineralization was examined for osteoblasts seeded in a porous scaffold composed of fused chitosan/nano-hydroxyapatite microspheres. Human fetal osteoblasts were cultured on composite and chitosan scaffolds for 21 days. On days 1, 4, 7, 14, and 21, total dsDNA, alkaline phosphatase, type I collagen, and osteocalcin production were measured. Total cellularity (measured by dsDNA), alkaline phosphatase, and type I collagen production were similar between the two scaffold groups. However, osteocalcin production occurred significantly earlier (day 7 vs. day 21) and was more than three times greater (0.0022 vs. 0.0068 ng/mL/ng DNA) on day 21 when osteoblasts were cultured on composite scaffolds. Osteocalcin is a marker of late osteoblastic differentiation and mineralized bone matrix formation. Therefore, the increase in osteocalcin production seen when cells were cultured on composite scaffolds may indicate that these scaffolds were superior to chitosan-only scaffolds in facilitating osteoblast mineralization. Composite scaffolds were also shown to be biocompatible and osteoconductive in a preliminary critical size rat calvarial defect study. These results demonstrate the potential of composite chitosan/nano-hydroxyapatite scaffolds to be used in bone tissue engineering. PMID- 19309241 TI - Infusion of mesenchymal stromal cells can aid hematopoietic recovery following allogeneic hematopoietic stem cell myeloablative transplant: a pilot study. AB - Mesenchymal stromal cells (MSCs) are important in the support of hematopoiesis. In this pilot study, we evaluated the safety and efficiency of donor-expanded MSC infusion after allogeneic hematopoietic stem cell transplantation (HSCT) in six patients with poor hematopoietic recovery. MSCs were infused without HSC and without conditioning at a dose of 1 x 10(6)/kg weight. Two patients displayed rapid hematopoietic recovery (days 12 and 21), and four patients showed no response. The two patients who showed hematopoietic recovery were in first complete remission (CR1) compared to the other heavily pretreated patients. There were no toxic side effects linked to MSC infusion. One patient developed cytomegalovirus (CMV) reactivation 12 days following the MSC infusion and died from CMV disease. We found that infusion of MSCs without HSC co-infusion can restore medullary function in some patients with poor hematopoietic recovery. Our data suggest that patients with a less damaged stroma could benefit from this approach. PMID- 19309242 TI - Heparin-induced anaphylactic and anaphylactoid reactions: two distinct but overlapping syndromes. AB - BACKGROUND: Heparin-induced anaphylactic and anaphylactoid reactions are of increasing clinical and scientific interest, particularly given the recent identification of a syndrome of heparin-induced anaphylaxis due to oversulfated chondroitin sulfate (OSCS), a contaminant in certain heparin preparations. However, heparin-induced anaphylactoid reactions also have been reported to be a consequence of immune-mediated heparin-induced thrombocytopenia (HIT). OBJECTIVE: To summarize the clinical features and pathophysiology of two distinct disorders, HIT-associated anaphylactoid reactions as well as anaphylaxis resulting from OSCS contaminated heparin. METHODS: We review literature describing these two types of heparin-induced anaphylactic and anaphylactoid reactions, and seek potential pathophysiologic links between them. RESULTS: Intravenous bolus heparin administered to patients with circulating 'HIT antibodies', usually as a result of recent heparin therapy, can produce anaphylactoid reactions, probably as a consequence of in vivo activation of platelets and, possibly, leukocytes. Affected patients often evince fever/chills, hypertension and/or acute respiratory compromise ('pseudo-pulmonary embolism'). In contrast, heparin induced anaphylaxis is caused by activation of the contact system, with formation of vasoactive kinins (bradykinin, des-arg(9)-bradykinin). This latter syndrome has been linked in an epidemic form to administration of OSCS-contaminated heparin; these reactions feature prominent hypotension and laryngeal edema. Hemodialysis patients are at increased risk for both syndromes. There is evidence that OSCS-contaminated heparin itself increases the risk of HIT compared with non contaminated heparin. CONCLUSION: Two distinct syndromes of heparin-induced anaphylaxis and anaphylactoid reactions exist. These seem to share certain epidemiologic features, given that OSCS-contaminated heparin can produce anaphylaxis through contact system activation but also could increase risk of HIT and HIT-associated anaphylactoid reactions. PMID- 19309243 TI - Drug safety profile of darbepoetin alfa for anemia of chronic kidney disease. AB - Anemia of chronic kidney disease due to deficiency of erythropoietin is common and has clinical consequences. Erythropoiesis stimulating agents including darbepoetin alfa (DA) are effective in correcting anemia. DA is generally well tolerated and has side effect profile similar to recombinant human erythropoietin. It has a long half-life permitting infrequent dosing. DA has been tested extensively in preclinical and clinical studies and significant experience has accumulated in clinical practice. Global safety profile of DA must consider recent data indicating worse survival, poor cardiovascular outcomes and thrombotic risks of targeting near normal hemoglobin levels and administering high doses of erythropoiesis stimulating agents. Strategies to achieve and maintain a reasonable, individualized target hemoglobin level with minimal variations in hemoglobin level are needed. PMID- 19309244 TI - Ifosfamide nephrotoxicity in children: a mechanistic base for pharmacological prevention. AB - The antineoplastic drug ifosfamide (IFO) in the treatment of solid tumors, particularly in children, is the cause of severe nephrotoxicity. Although it is a potent and effective chemotherapeutic agent, the associated nephrotoxicity has a serious impact on the health and the quality of life of exposed children. The toxic metabolite of IFO thought to be responsible for IFO-induced kidney damage is chloroacetaldehyde (CAA). Those suffering from nephrotoxicity typically develop tubular and glomerular toxicities, with the most severe form being Fanconi's syndrome. As the mode of toxicity of CAA seems to be primarily owing to oxidative stress, the use of antioxidants as a protective measure for the kidneys is a promising strategy. In this review, we highlight recent research that supports the local renal production of CAA as the proximate cause of IFO-induced nephrotoxicity with age as an important risk factor, those under the age of three being the most vulnerable. Most importantly, we focus on the potential advantages of the antioxidant N-acetylcysteine owing to both its antioxidant properties and its current use clinically in pediatrics. PMID- 19309245 TI - Antiplatelet therapy and coronary artery bypass graft surgery: perioperative safety and efficacy. AB - BACKGROUND: Antiplatelet therapy is critical in the management of coronary artery disease. For patients undergoing coronary artery bypass graft surgery (CABG), controversy remains regarding the safety of preoperative antiplatelet therapy and the optimal postoperative antiplatelet regimen to maintain graft patency and reduce ischemic complications. OBJECTIVE: The goal of this systematic review is to evaluate the risks and benefits of preoperative aspirin and clopidogrel therapy, to identify the optimal timing and dose of aspirin following CABG, and to assess the role of postoperative clopidogrel therapy. METHODS: A systematic review was performed to retrieve relevant articles from the Medline database published between 1978 and 2008. RESULTS: With respect to aspirin, the published data suggest that it should be held for 7 - 10 days preoperatively in patients undergoing elective CABG. However, in the current era of routine antifibrinolytic therapy and the high prevalence of patients with extensive atherosclerotic disease, this practice has been put into question. Clopidogrel should be held for at least 5 days before CABG to avoid perioperative bleeding complications. Following surgery, extensive evidence supports the use of aspirin, in doses of 100 - 325 mg daily, to be administered in 48 h postoperatively and continued indefinitely. Less is known regarding the use of clopidogrel following CABG, although it is now recommended as postoperative antiplatelet therapy in patients with recent acute coronary syndromes. CONCLUSION: Despite > 30 years of experience with antiplatelet agents during CABG, questions remain regarding their perioperative safety and efficacy. The results of continuing randomized controlled trials should further clarify the role of perioperative aspirin and clopidogrel therapy and help redefine the modern antiplatelet management of coronary artery bypass patients. PMID- 19309246 TI - Recognition and management of drug-induced cytopenias: the example of idiosyncratic drug-induced thrombocytopenia. AB - BACKGROUND: Several hundred drugs, toxins and herbs have been reported to cause blood abnormalities, and drugs account for 20 - 40% of all instances of cytopenias. OBJECTIVE: In the present paper, we report and discuss the recognition and management of moderate to severe idiosyncratic drug-induced thrombocytopenia. METHODS: A bibliographic search was performed on the PubMed database of the US National Library of Medicine for articles published from January 1990 to November 2008. RESULTS/CONCLUSIONS: Moderate to severe idiosyncratic drug-induced thrombocytopenia (platelet count < 100 x 10(9)/l) is a relatively rare and potentially serious disorder. The origin may be myelosuppression or peripheral, owing to either the consumption of platelets or their immune-mediated destruction. The most common molecules responsible are heparins, quinidine, sulfonamides and gold salts. Clinically, the most classical symptom is a typical pattern of bleeding of variable intensity depending on the severity of thrombocytopenia and the molecule involved. Immune-mediated thrombocytopenia induced by heparin (type II) is more often associated with thrombotic events. The diagnosis is based on medical history and a set of clinical criteria, which also specify the level of imputability. Although the role of serological tests is not well established, they seem particularly valuable in some situations in which differential diagnosis is difficult or in type II heparin-induced thrombocytopenia. The treatment includes discontinuation of the suspected drug, and symptomatic measures that depend on the severity of clinical symptoms. PMID- 19309247 TI - Fluoropyrimidine-associated cardiotoxicity: revisited. AB - BACKGROUND: The syndrome of 5-fluorouracil (5-FU)-associated cardiotoxicity remains poorly defined. PATIENTS AND METHODS: We performed a literature review (1969 - 2007) and compiled data derived from 377 evaluable cases out of 448 reported cases. RESULTS: Patient age ranged from 14 to 86 years. Of the patients 65% were 55 years old and the male:female ratio was 1.5:1. The most commonly treated tumors were gastrointestinal (60%), head and neck (22%) and breast (4%). Of the patients 14% had a history of heart disease whereas cardiac risk factors were found in 37%. Mode of administration included: continuous infusion (72%); bolus (22.5%); intermediate infusion (3%); oral (2%); and intraperitoneal (1 patient). The dosages of 5-FU used were < 750 mg/m(2)/day (36%), 751 - 999 (16%), 1,000 (26%), 1,001 - 1,499 (4%) and 1,500 (16%). Of the patients 54% received 5 FU in combination with other chemotherapeutic agents (cisplatin 44%) whereas 51% received 5-FU alone or with leucovorin. Only 4% patients had undergone previous or concomitant radiation therapy to the mediastinum. Of cardiac incidents that happened 69% were seen during or within 72 h of the first cycle of 5-FU. Angina occurred in 45% of patients whereas myocardial infarction was seen in 22%, arrhythmias in 23, acute pulmonary edema in 5, cardiac arrest and pericarditis in 1.4 and heart failure in 2. Electro-cardiographic evidence of ischemia or ST-T changes were recorded in 69% of patients, but abnormal cardiac enzymes were found in only 12%. The cardiac symptoms were reproducible in 47%, including in one patient subsequently treated with 5-FU p.o. Symptoms were also elicited when the same patients were treated with lower doses or different schedules. Of the patients 68% responded to conservative anti-anginal therapy, although prophylactic coronary vasodilators had limited efficacy. Overall, 8% of patients showing cardiotoxicity on 5-FU administration died. Furthermore, 13% reexposed to 5-FU died. CONCLUSIONS: Our review suggests that 5-FU cardiotoxicity is an infrequent but real phenomenon that is independent of dose and may be related to a continuous infusion schedule. The presence of cardiac risk factors is not predictive. Patients should be observed closely and 5-FU administration discontinued if cardiac symptoms develop. A rechallenge with 5-FU should be reserved only for those patients in whom there is no reasonable alternative therapy and should be performed in the setting of aggressive prophylaxis and close monitoring. PMID- 19309248 TI - The safety of plasma-derived von Willebrand/factor VIII concentrates in the management of inherited von Willebrand disease. AB - Until the mid-80s, cryoprecipitate has been the mainstay of treatment of patients with von Willebrand disease who were unresponsive to desmopressin. The advent of virally-inactivated factor VIII (FVIII) concentrates containing von Willebrand factor (VWF), originally devoted to hemophiliacs, provided a better therapeutic approach to von Willebrand disease. These VWF/FVIII concentrates were introduced in clinical practice after the positive results obtained in several prospective and retrospective clinical studies. They are safe and can be suitable also for home treatment. Allergic or anaphylactic reactions are limited to the rare patients with deletions of VWF gene. In repeated infusions during surgery, the dosage and timing of administration should be planned to keep FVIII below 150 - 200 U/dl to avoid any possible risk of thrombosis. PMID- 19309249 TI - Safety of 17D derived yellow fever vaccines. AB - BACKGROUND: Yellow fever, a mosquito-borne viral haemorrhagic fever, is one of the most lethal viral diseases. At present, an efficient vaccine for prevention is available, but may cause serious adverse events. METHODS: The authors review the up-to-date knowledge for serious adverse events of the yellow fever vaccine (YFSAE): hypersensitivity reactions, neurotropic and viscerotropic syndromes. RESULTS: The incidence of YFSAE has been associated with increasing age and thymus disorders, but still a number of cases with any risk factor remain unexplained, which suggests that other factors, from the virus or the host, could be involved in the appearance of these postvaccinal adverse events. CONCLUSIONS: YFSAE are uncommon but must be prevented. Further research on the virus-host immune response is needed to have a better understanding of the basis for the appearance of these severe side effects after vaccination. Vaccination should be limited to people with a true risk of exposure to wild-type yellow fever virus. PMID- 19309250 TI - Safety Pharmacology Society: 8th annual meeting. AB - Of the numerous topics discussed during the eighth annual meeting of the Safety Pharmacology (SP) Society the author identified the following key topics: i) the impact of hERG (human ether-a-go-go related gene) channel data on drug development, ii) safety evaluation of biological products, iii) opportunities and expectations from SP, iv) human stem cell derived cardiomyocytes for safety assessment, v) role of cellular calcium pathways in drug-induced arrhythmias, vi) collaboration initiatives for finding solutions to cardiac repolarisation and other recognised SP risks, vii) Pharma and FDA perspectives on ICH S7B guideline, viii) joint PhRMA-FDA dialogue on drug abuse potential assessment, ix) frontloading SP strategies for mitigating non-clinical and clinical attrition; and x) approaches to measure non-clinical assay predictability for human outcome. The establishment of consortia to accelerate the solution of critical SP issues and the implementation of Frontloading SP or Exploratory SP strategies for an early selection of safe clinical candidates are promising avenues for consolidating SP as an indispensable drug development discipline and for transforming established regulatory SP investigations into a risk-known exercise. PMID- 19309251 TI - Efficacy and safety of therapy with metformin plus pioglitazone in the treatment of patients with type 2 diabetes: a double-blind, placebo-controlled, clinical trial. AB - OBJECTIVE: To assess the efficacy and safety of combination therapy with pioglitazone and metformin in Japanese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: During a 12-week observation period 236 patients were treated with metformin 500 or 750 mg/day. 169 patients with a confirmed HbA(1c) level >or= 6.5% were randomized (stratified according to metformin dosage) to receive pioglitazone 15 mg/day for 12 weeks then increased to 30 mg/day for a further 16 weeks (n = 83), or placebo (n = 86). Outcome measures included HbA(1c), fasting blood glucose (FBG), percentage of patients achieving HbA(1c) < 6.5%, lipid profile, and other metabolic parameters. RESULTS: Mean HbA(1c) was reduced by 0.67% in patients receiving pioglitazone plus metformin versus an increase of 0.25% in those receiving metformin alone (p < 0.0001). After 8 weeks' treatment and until the end of the study, HbA(1c) was significantly lower with pioglitazone plus metformin and more patients in this group achieved an HbA(1c) < 6.5% (38.6% vs. 8.1%; p < 0.0001). FBG was also reduced by a significantly greater amount in patients receiving pioglitazone plus metformin compared with metformin monotherapy (-20.5 vs. 1.9 mg/dl; p < 0.0001). Combination therapy was associated with significantly increased HDL-cholesterol, total cholesterol, and adiponectin, and significantly decreased levels of fasting insulin, free fatty acids, and homeostasis model assessment insulin resistance (HOMA-R) compared with metformin monotherapy. Overall, combination therapy and monotherapy were equally well tolerated and the incidence of adverse effects 'possibly' related to therapy was 15.7% and 11.6% (p = 0.505), respectively. Edema occurred slightly more often in the combination group (6.0 vs. 1.2%). CONCLUSION: Pioglitazone plus metformin significantly improved glycemic control (HbA(1c) and FBG), and markers associated with increased insulin resistance and cardiovascular risk compared with metformin monotherapy. CLINICAL TRIAL REGISTRATION NUMBER: UMIN 000001110. PMID- 19309252 TI - Effect of sitting position on respiratory status in preterm infants. AB - OBJECTIVE: To evaluate whether using a sitting-type car safety seats for preterm infants is advisable. PATIENTS AND METHODS: A total of 65 preterm infants underwent polysomnography in the supine and sitting positions. The infants with <95% of SpO(2) were assigned to the desaturation (DS) group. Their backgrounds, breathing patterns, and breathing types were analyzed. RESULTS: Of the 65 cases, 18 were assigned to the DS group. No significant differences were observed between the DS and non-DS groups in their background characteristics. Of the 18 DS cases, 15 were included in the non-obstructive group (8, periodic breathing; 6, tachypnea; 1, irregular breathing). CONCLUSIONS: Immaturity of the respiratory center and decompensation for the respiratory changes caused by the sitting position were suspected to cause DS in infants. Sitting-type car safety seats should be used with caution for preterm infants, and all preterm infants need to be screened by polysomnographic examination in the sitting position. PMID- 19309253 TI - Temporal trends in pregnancy weight gain and birth weight in Bavaria 2000-2007: slightly decreasing birth weight with increasing weight gain in pregnancy. AB - AIMS: To assess temporal trends in birth weight and pregnancy weight gain in Bavaria from 2000 to 2007. METHODS: Data on 695,707 mother and infant pairs (singleton term births) were available from a compulsory reporting system for quality assurance, including information on birth weight, maternal weight at delivery and at booking, maternal smoking, age, and further anthropometric and lifestyle factors. Pregnancy weight gain was defined as: weight prior to delivery minus weight at first booking minus weight of the newborn. RESULTS: Although mean weight gain during pregnancy increased considerably from 10.10 to 10.73 kg in seven years, the mean birth weight in mature singletons decreased slightly from 3433 to 3414 g. These trends could not be explained by concurrent changes in the rates of primiparity, smoking and gestational diabetes. CONCLUSIONS: These German data confirm an increased weight gain during pregnancy with adjustment for potential confounders. PMID- 19309254 TI - A new device to facilitate the chorion villus sampling. PMID- 19309255 TI - Correct definition of pentalogy of Cantrell. PMID- 19309256 TI - Downstream targets and intracellular compartmentalization in Nox signaling. AB - Reactive oxygen species (ROS) have become recognized for their role as second messengers in a multitude of physiologic responses. Emerging evidence points to the importance of the NADPH oxidase family of ROS-producing enzymes in mediating redox-sensitive signal transduction. However, a clear paradox exists between the specificity required for signaling and the nature of ROS as both diffusible and highly reactive molecules. We seek to understand the targets and compartmentalization of the NADPH oxidase signaling to determine how NADPH oxidase-derived ROS fit into established signaling paradigms. Herein we review recent data that link cellular NADPH oxidase enzymes to ROS signaling, with a particular focus on the mechanism(s) involved in achieving signaling specificity. PMID- 19309257 TI - Redox regulation, NF-kappaB, and atrial fibrillation. AB - Atrial fibrillation (AF) is the most common clinically encountered abnormal heart beat. It is associated with an increased risk of stroke and symptoms of heart failure. Current therapies are directed toward controlling the rate of ventricular activation and preventing strokes through anticoagulation. Attempts at suppressing the arrhythmia are often ineffective, in part because the underlying pathogenesis is poorly understood. Recently, structural and electrical remodeling has been shown to occur during AF. These changes involve alterations in gene regulation and help perpetuate the arrhythmia. Some signals for remodeling are have been identified. Moreover, AF is associated with oxidative stress, and this redox imbalance may contribute to the altered gene regulation. One likely mediator of this change in transcriptional regulation is the redox sensitive transcription factor, nuclear factor-kappaB (NF-kappaB). Recently, NF kappaB has been shown to downregulate transcription of the cardiac sodium channel in response to oxidative stress. NF-kappaB may contribute to the regulation of other ion channels, transcription factors, or splicing factors altered in AF and may represent a therapeutic target in AF management. PMID- 19309258 TI - Molecular mechanisms responsible for the atheroprotective effects of laminar shear stress. AB - The endothelium lining the inner surface of blood vessels of the cardiovascular system is constantly exposed to hemodynamic shear stress. The interaction between endothelial cells and hemodynamic shear stress has critical implications for atherosclerosis. Regions of arterial narrowing, curvatures, and bifurcations are especially susceptible to atherosclerotic lesion formation. In such areas, endothelial cells experience low, or oscillatory, shear stress. Corresponding changes in endothelial cell structure and function make them susceptible to the initiation and development of atherosclerosis. In contrast, blood flow with high laminar shear stress activates signal transductions as well as gene and protein expressions that play important roles in vascular homeostasis. In response to laminar shear stress, the release of vasoactive substances such as nitric oxide and prostacyclin decreases permeability to plasma lipoproteins as well as the adhesion of leukocytes, and inhibits smooth muscle cell proliferation and migration. In summary, different flow patterns directly determine endothelial cell morphology, metabolism, and inflammatory phenotype through signal transduction and gene and protein expression. Thus, high laminar shear stress plays a key role in the prevention of atherosclerosis through its regulation of vascular tone and long-term maintenance of the integrity and function of endothelial cells. PMID- 19309259 TI - Role of the unfolded protein response regulator GRP78/BiP in development, cancer, and neurological disorders. AB - GRP78/BiP is a major endoplasmic reticulum (ER) chaperone protein critical for protein quality control of the ER, as well as controlling the activation of the ER-transmembrane signaling molecules. Through creation of mouse models targeting the Grp78 allele, the function of GRP78 in development and disease has been investigated. These led to the discovery that GRP78 function is obligatory for early embryonic development. However, in adult animals, GRP78 is preferably required for cancer cell survival under pathologic conditions such as tumor progression and drug resistance. The discovery of surface localization of GRP78 in cancer cells reveals potential novel function, interaction with cell-surface receptors, and possible therapeutic implications. Mouse models also reveal that GRP78 controls maturation and secretion of neuronal factors for proper neural migration and offers neuroprotection. PMID- 19309260 TI - NADPH oxidase 1 deficiency alters caveolin phosphorylation and angiotensin II receptor localization in vascular smooth muscle. AB - The superoxide-generating NADPH oxidase NOX1 is thought to be involved in signaling by the angiotensin II-receptor AT1R. However, underlying signaling steps are poorly understood. In this study, we investigated the effect of AngII on aortic smooth muscle from wild-type and NOX1-deficient mice. NOX1-deficient cells showed decreased basal ROS generation and did not produce ROS in response to AngII. Unexpectedly, AngII-dependent Ca(2+) signaling was markedly decreased in NOX1-deficient cells. Immunostaining demonstrated that AT1R was localized on the plasma membrane in wild-type, but intracellularly in NOX1-deficient cells. Immunohistochemistry and immunoblotting showed a decreased expression of AT1R in the aorta of NOX1-deficient mice. To investigate the basis of the abnormal AT1R targeting, we studied caveolin expression and phosphorylation. The amounts of total caveolin and of caveolae were not different in NOX1-deficient mice, but a marked decrease occurred in the phosphorylated form of caveolin. Exogenous H(2)O(2) or transfection of a NOX1 plasmid restored AngII responses in NOX1 deficient cells. Based on these findings, we propose that NOX1-derived reactive oxygen species regulate cell-surface expression of AT1R through mechanisms including caveolin phosphorylation. The lack cell-surface AT1R expression in smooth muscle could be involved in the decreased blood pressure in NOX1-deficient mice. PMID- 19309261 TI - Small-molecule NOX inhibitors: ROS-generating NADPH oxidases as therapeutic targets. AB - NOX NADPH oxidases are electron-transporting membrane enzymes whose primary function is the generation of reactive oxygen species (ROS). ROS produced by NOX enzymes show a variety of biologic functions, such as microbial killing, blood pressure regulation, and otoconia formation. Strong evidence suggests that NOX enzymes are major contributors to oxidative damage in pathologic conditions. Blocking the undesirable actions of NOX enzymes, therefore, is a therapeutic strategy for treating oxidative stress-related pathologies, such as ischemia/reperfusion tissue injury, and neurodegenerative and metabolic diseases. Most currently available NOX inhibitors have low selectivity, potency, and bioavailability, precluding a pharmacologic demonstration of NOX as therapeutic targets in vivo. This review has two main purposes. First, we describe a systematic approach that we believe should be followed in the search for truly selective NOX inhibitors. Second, we present a critical review of small-molecule NOX inhibitors described over the last two decades, including recently published patents from the pharmaceutical industry. Structures, activities, and in vitro/in vivo specificity of these NOX inhibitors are discussed. We conclude that NOX inhibition is a pertinent and promising novel pharmacologic concept, but that major efforts will be necessary to develop specific NOX inhibitors suited for clinical application. PMID- 19309263 TI - NOX enzymes in the central nervous system: from signaling to disease. AB - Oxidative stress has been implicated in the pathogenesis of neurologic and psychiatric diseases. The brain is particularly vulnerable to oxidative damage due to high oxygen consumption, low antioxidant defense, and an abundance of oxidation-sensitive lipids. Production of reactive oxygen species (ROS) by mitochondria is generally thought to be the main cause of oxidative stress. However, a role for ROS-generating NADPH oxidase NOX enzymes has recently emerged. Activation of the phagocyte NADPH oxidase NOX2 has been studied mainly in microglia, where it plays a role in inflammation, but may also contribute to neuronal death in pathologic conditions. However, NOX-dependent ROS production can be due to the expression of other NOX isoforms, which are detected not only in microglia, but also in astrocytes and neurons. The physiologic and pathophysiologic roles of such NOX enzymes are only partially understood. In this review, we summarize the present knowledge about NOX enzymes in the central nervous system and their involvement in neurologic and psychiatric diseases. PMID- 19309262 TI - Novel role of NADPH oxidase in angiogenesis and stem/progenitor cell function. AB - Neovascularization is involved in normal development and wound repair as well as ischemic heart disease and peripheral artery disease. Both angiogenesis and vasculogenesis [de novo new vessel formation through mobilization of stem/progenitor cells from bone marrow (BM) and their homing to the ischemic sites] contribute to the formation of new blood vessels after tissue ischemia. Angiogenesis is dependent on cell proliferation, migration, and capillary tube formation in endothelial cells (ECs). Stem/progenitor cells have been used for cell-based therapy to promote revascularization after peripheral or myocardial ischemia. Excess amounts of reactive oxygen species (ROS) are involved in senescence and apoptosis of ECs and stem/progenitor cells, causing defective neovascularization. ROS at low levels function as signaling molecules to mediate cell proliferation, migration, differentiation, and gene expression. NADPH oxidase is one of the major sources of ROS in ECs and stem/progenitor cells, and is activated by various growth factors, cytokines, hypoxia, and ischemia. ROS derived from NADPH oxidase play an important role in redox signaling linked to angiogenesis ECs, as well as stem/progenitor cell mobilization, homing, and differentiation, thereby promoting neovascularization. Understanding these mechanisms may provide insight into NADPH oxidase and its mediators as potential therapeutic targets for ischemic heart and limb disease. PMID- 19309265 TI - Long-term cyclic strain downregulates endothelial Nox4. AB - Endothelial cells in vivo are constantly exposed to mechanical forces such as cyclic strain. In endothelial cells, Nox4-containing NAD(P)H oxidase complexes have been identified as major sources of superoxide anion (.O(2)(-)) formation. In this study, we analyzed the effect of cyclic strain on endothelial ROS formation by electron paramagnetic resonance spectroscopy, cytochrome c assay, and dihydroethidium fluorescence, on NO formation by Griess reaction and on gene expression by RT-PCR and Western blot. Primary cultures of human umbilical vein endothelial cells were exposed to 2-18% cyclic strain for up to 24 h using the Flexercell system. Long-term application of 5-12% cyclic strain downregulated Nox4 expression and ROS formation in a time-dependent manner. Downregulation of Nox4 was further confirmed by promoter analysis using dual-luciferase assay. Cu/Zn SOD, MnSOD, and catalase expression was decreased after application of chronic 12% cyclic strain. In contrast, endothelial NO formation and eNOS were increased by cyclic strain. Strain-dependent Nox4 downregulation was abolished by eNOS inhibition with L-NAME. In conclusion, physiological levels of cyclic strain downregulate Nox4 expression and superoxide anion formation. This novel mechanism might contribute to a vasoprotective balance between NO and superoxide anions in response to physiological mechanical stimulation of endothelial cells. PMID- 19309264 TI - A role for copper in the toxicity of zinc-deficient superoxide dismutase to motor neurons in amyotrophic lateral sclerosis. AB - In the 16 years since mutations to copper, zinc superoxide dismutase (SOD1) were first linked to familial amyotrophic lateral sclerosis (ALS), a multitude of apparently contradictory results have prevented any general consensus to emerge about the mechanism of toxicity. A decade ago, we showed that the loss of zinc from SOD1 results in the remaining copper in SOD1 to become extremely toxic to motor neurons in culture by a mechanism requiring nitric oxide. The loss of zinc causes SOD1 to become more accessible, more redox reactive, and a better catalyst of tyrosine nitration. Although SOD1 mutant proteins have a modestly reduced affinity for zinc, wild-type SOD1 can be induced to lose zinc by dialysis at slightly acidic pH. Our zinc-deficient hypothesis offers a compelling explanation for how mutant SOD1s have an increased propensity to become selectively toxic to motor neurons and also explains how wild-type SOD1 can be toxic in nonfamilial ALS patients. One critical prediction is that a therapeutic agent directed at zinc-deficient mutant SOD1 could be even more effective in treating sporadic ALS patients. Although transgenic mice experiments have yielded contradictory evidence to the zinc-deficient hypothesis, we will review more recent studies that support a role for copper in ALS. A more careful examination of the role of copper and zinc binding to SOD1 may help counter the growing disillusion in the ALS field about understanding the pathological role of SOD1. PMID- 19309266 TI - Genetic counseling: a medical approach. AB - The expanding role of genetics in almost every clinical condition is associated with the increasing development of new genetic and molecular test, and implementation of these tests is becoming well integrated into medical practice. Therefore, primary care providers, family doctors, pediatricians, and other specialists involved in the care of patients and families affected by genetic conditions will be largely concerned with both genetic tests and genetic counseling. That process may be considered in several stages: collecting genetic information and pedigree drawing; making or validating the diagnosis; estimating occurrence and recurrence risk; communicating clinical information; and supporting the family to reach a decision and take appropriate action. However, the term "genetic counseling" is often used to describe the entire approach to patients and families with genetic conditions, sometimes being restricted to the communication and psychotherapeutic process. Consequently, the separation of clinical from supportive process had an important role in most definitions of genetic counseling. Should all those aspects be considered parts of a single process? This is the main question that will be discussed hereafter, under the hypothesis that the synthesis of all those steps should be required to approach any genetic conditions. In that view, rethinking of genetic counseling from a medical point of view should be seen as an important subject for discussion, and might contribute to bring together clinical and nonmedical aspects. PMID- 19309267 TI - Psychosocial aspects of DNA testing for hereditary hemochromatosis in at-risk individuals: a systematic review. AB - AIM: To review the psychosocial benefits and harms of DNA testing for HFE-related hereditary hemochromatosis (HH) in at-risk individuals. BACKGROUND: HH is a common genetic disease in people of European descent. DNA-based predisposition testing is used for diagnosis or in the context of family testing, but there are concerns about potential psychosocial consequences. METHODS: Fifteen electronic databases (including Medline and Cochrane) were searched from inception to April 2007 to identify any quantitative or qualitative primary research that considered DNA testing of individuals considered at-risk of HH and reported psychosocial outcomes. Inclusion criteria, data extraction, and quality assessment were undertaken by standard methodology. RESULTS: Three observational studies met the inclusion criteria of the review; each had methodological limitations. On receipt of test results, anxiety levels fell or were unchanged; general health-related quality-of-life outcomes improved in some aspects, or were unchanged with respect to pretest result values. Outcomes were not reported separately for those referred for diagnosis and those with family history of HH. Results suggest that genetic testing for HH in at-risk individuals is accompanied by few negative psychosocial outcomes. CONCLUSION: The evidence on the psychosocial aspects of DNA testing for HH in at-risk individuals is limited. Further research might be required if other factors influencing the natural history of the disease phenotype are identified. PMID- 19309268 TI - Cornelia de Lange syndrome: a case study. AB - Cornelia de Lange syndrome (CDLS) is a relatively common multiple congenital anomaly/mental retardation disorder with an unknown genetic and molecular pathogenesis. The essential features of this developmental malformation syndrome are retardation in growth, developmental delay, various structural limb abnormalities, and distinctive facial features. Most cases are sporadic and are thought to result from a new dominant mutation. Consequently, hypotheses regarding the pathogenetic mechanisms underlying the two distinct phenotypes, classic and mild, are purely speculative. The recent discovery of molecular techniques and identification of the NIPBL gene has allowed etiologic diagnosis of this disorder. In this article, we describe a patient with CDLS in whom conventional cytogenetics, fluorescence in situ hybridization, and NIPBL gene mutation analysis determined an etiologic diagnosis, providing precise genetic counseling and facilitated the family to make an evidence-based decision for conception and also alleviated the extreme degree of anxiety associated with the thought of having a second child in this set of circumstances. PMID- 19309269 TI - A multiplexed ARMS-PCR approach for the detection of common MECP2 mutations. AB - Rett syndrome (RTT), an X-linked dominant neurodevelopmental disorder, is caused mainly by de novo mutations in the methyl-CpG-binding protein 2 gene (MECP2). Although more than 200 different MECP2 mutations have been identified throughout the gene, 7 of those (p.R133C, p.T158M, p.R168X, p.R255X, p.R270X, p.R294X, and p.R306C) account for up to two-thirds of pathogenic mutations in RTT patients. A rapid and efficient screening strategy for these mutations can be used as a preliminary step for genetic diagnosis of RTT. The current protocols used for this purpose are of high cost and require special equipment. We have designed a simpler multiplex amplification refractory mutation system (ARMS)-PCR strategy that allows identification of these common MECP2 mutant alleles in four PCR reactions. The assay was tested in 14 RTT patients who were previously genotyped using PCR-restriction fragment length polymorphism and DNA sequencing. A complete concordance was observed between the results of the two methods. The multiplex ARMS-PCR does not require any special equipment, and it provides rapid, reproducible, and cost-effective detection of common MECP2 mutations. The assay can be carried out efficiently in a standard molecular genetics laboratory and suitable as a preliminary screen for all patients with RTT diagnosis. PMID- 19309270 TI - Characteristics of dystrophin gene mutations among Chinese patients as revealed by multiplex ligation-dependent probe amplification. AB - AIMS: To verify whether dystrophin gene mutations among Chinese patients feature different types and frequencies from other populations. METHODS: Multiplex ligation-dependent probe amplification (MLPA) in combination with multiplex PCR (mPCR) and/or short tandem repeat (STR)-based linkage analysis were applied in a large series of Chinese families affected with Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy (BMD). There were a total of 19 cases seeking prenatal diagnosis during their second pregnancies. RESULTS: Of the 59 family trios (51 with DMD and 8 with BMD), 40 were found to have carried various mutations of the dystrophin gene. In addition to deletions and duplications within the mutational hotspots identified by both methods, 10 mutations missed by mPCR were detected by MLPA, among which at least 3 were of rare types. Combined MLPA and linkage analysis also achieved prenatal diagnoses in all of the 19 amniocentesis samples. CONCLUSIONS: Mutations of dystrophin gene among Chinese patients showed a diverse spectrum, with similarity to as well as discrepancies from other populations. For the comprehensive coverage of all exons of the dystrophin gene, MLPA should be the method of choice for initial screening of DMD/BMD patients. When combined with STR-based analysis, it can achieve diagnosis in as much as 70-80% of all referred cases. PMID- 19309271 TI - The Italian scheme of External Quality Assessment for beta-thalassemia: genotyping and reporting results and testing strategies in a 5-year survey. AB - The Italian scheme of External Quality Assessment for beta-thalassemia started in 2001 as part of a project twice funded by the Italian Ministry of Health and coordinated by the Istituto Superiore di Sanita. To date, five trials have been performed (2001-2004 and 2006). The aim of the Italian scheme is to help public laboratories in improving and reaching a high standard of quality when performing a molecular test. Many laboratories took part in the 5-year project, and their participation was constant during the whole period. The aims of this paper are to describe the genotyping and reporting results as well as focusing on the techniques and the testing strategies adopted to detect mutations. Almost 99% of the alleles analyzed were correctly detected by laboratories, while 0.33% of the analyses gave a wrong result. Reverse dot blot was the most used technique, and it was always used in the strategies adopted by laboratories to detect mutations. The reports sent by laboratories showed incompleteness and heterogeneity; thus, a new model for written reports has been introduced since 2004. It will be interesting to monitor the effects of the reporting model and the output of this educational action in the future. PMID- 19309272 TI - A simple multiplex real-time PCR methodology for the SMN1 gene copy number quantification. AB - Spinal muscular atrophy (SMA) is an autosomal recessive disease caused, in about 95% of SMA cases, by homozygous deletion of the survival motor neuron 1 (SMN1) gene or its conversion to the highly homologous SMN2 gene. The molecular diagnosis of SMA is usually carried out by a PCR-Restriction fragment length polymorphism (RFLP) approach. However, this approach is not useful for identification of healthy deletion carriers. TaqMan technology is one of the most reliable and widely adopted techniques for the SMN1 copy number evaluation. However, several limitations of this technique have been described. Particularly, DNA extraction methods and accurate template quantification have been shown to be critical for reliable results. In this work, we set up a reliable, highly reproducible, and easy-to-perform TaqMan technology-based protocol to obtain the SMN1 gene copy number assessment. We demonstrate that PCR amplification of both target gene and reference gene in the same reaction mix, instead of separated mixes, greatly reduces reported criticisms of simplex TaqMan technology. The multiplex real-time PCR we describe allows interlaboratory samples and data exchange, without the need to equalize the DNA isolation technique. Further, the protocol described below requires fewer replica tests than the simplex methodology does, leading to reduced overall cost for the diagnostic assay. PMID- 19309273 TI - Novel EXT1 and EXT2 mutations in hereditary multiple exostoses families of Indian origin. AB - BACKGROUND: Hereditary multiple exostosis (HME) is an autosomal dominant bone disorder, characterized by short stature and the presence of multiple benign tumors mainly at the ends of long bones. HME is genetically heterogeneous with two known genes on 8q24 (EXT1) and 11p11 (EXT2), and a third minor locus mapped to 19p (EXT3). The majority of EXT1 and EXT2 mutations result in premature protein truncation and loss of function. MATERIALS AND METHODS: We analyzed two autosomal dominant HME families of Indian origin. Linkage analysis using fluorescently labeled microsatellite markers at the candidate gene regions was performed. Mutation analysis was carried out by bidirectional sequencing of purified PCR products. RESULTS: We found linkage in one family to EXT1 and in the other family to EXT2. Mutation screening in the EXT1 gene revealed a novel frameshift mutation, a single base deletion in exon 1 (c.142delC). This mutation segregated in all affected members and was absent in the unaffected family members and 60 unrelated controls. In the second family, a previously unreported stop mutation, the substitution c.817C>T, was observed in the EXT2 gene in all affected members and in none of the unaffected family members and 90 unrelated controls. CONCLUSIONS: Our findings expand the mutation spectrum of EXT1 and EXT2 and highlight the genetic and phenotypic heterogeneity of HME. PMID- 19309274 TI - Uptake, time course, and predictors of risk-reducing surgeries in BRCA carriers. AB - INTRODUCTION AND AIMS: For women who carry BRCA mutations, risk-reducing surgeries are an option to decrease breast and ovarian cancer risk. This study aims to determine the uptake, time course, and predictors of risk-reducing mastectomy (RRM) and risk-reducing salpingo-oophorectomy (RRSO) in BRCA carriers. RESULTS: In 272 female carriers, followed for a median of 3.7 years, 23% of those eligible chose RRM, and 51% percent chose RRSO. Among BRCA carriers who chose these procedures, median time to both RRM and RRSO was approximately 4 months after learning of BRCA-positive results. Predictors of RRM were as follows: age below 60 years (hazard ratio 1.8, p=0.04), prior breast cancer (hazard ratio 2.4, p=0.0004), and RRSO (hazard ratio 7.2, p<0.0001). Predictors of RRSO were as follows: age below 60 years (hazard ratio 3.6, p=0.006), prior breast cancer (hazard ratio 1.8, p=0.002), and RRM (hazard ratio 5.4, p<0.0001). CONCLUSIONS: Many women who undergo BRCA testing use these results to make clinical decisions; those who choose risk-reducing surgeries typically do so within months of receiving BRCA-positive results. Predictors of risk-reducing surgery uptake include the following: age below 60 years, prior breast cancer, and utilization of another risk-reducing surgery. Future research directions include examining other preventive and screening options in BRCA carriers as well as studying motivations for choosing or declining risk-reducing surgeries. PMID- 19309275 TI - Potential consumers' attitudes toward psychiatric genetic research and testing and factors influencing their intentions to test. AB - Psychiatric genetic research brings on the possibility of psychiatric genetic testing. The optimal and responsible utilization of genetic testing depends on knowledge of the potential consumers' attitudes and expectations regarding testing. The aim of this study was to assess potential consumers' attitudes and expectations toward psychiatric genetics and factors influencing their intentions to test. A questionnaire constructed to assess attitudes and intentions toward psychiatric genetic testing was mailed or given in person to individuals participating in different genetic studies aiming at identifying genes predisposing for mental illness. A total of 397 persons diagnosed with major depression, bipolar disorder, schizophrenia, or anxiety disorder participated in the survey. A large majority of the sample expressed an intention for themselves and their children to participate in psychiatric genetic testing. Support for prenatal testing was considerably less strong. A large minority expressed intention to test regardless of treatment possibilities. Intentions to test were positively associated with being a parent, trust in researchers, and expecting to feel better prepared for fighting the disorder when knowing of the presence of risk genes. Intentions were negatively associated with the fear of psychiatric genetic research bringing on too many difficult choices and fearing not to be able to cope with the results of a psychiatric genetic test. These results indicate that psychiatric genetic testing is not just perceived as a way to better treatment. Other expectations may motivate testing even though the clinical validity of the test is poor. PMID- 19309276 TI - The association of methylation in the promoter of APC and MGMT and the prognosis of Taiwanese CRC patients. AB - AIMS: The purpose of this study was to investigate the association of methylation in the promoter regions of adenomatous polyposis coli (APC) and O(6) methylguanine-DNA methyltransferase (MGMT) and the survival of Taiwanese colorectal cancer (CRC) subjects who received 5-fluorouracil (5-FU) adjuvant chemotherapy. RESULTS: DNA isolated from tumor tissue of 117 CRC subjects was analyzed for the existence of methylation in the promoter regions of APC and MGMT by methylation-specific PCR. Various characteristics of the 117 subjects were recorded and used in the Cox proportional-hazard model analyses. Methylation in the promoter region is 62.4% (73/117) for APC and 60.7% (71/117) for MGMT in our CRC patients. Subjects presenting methylation in the APC promoter demonstrate significantly lower hazards for all causes of death (hazard ratios=0.378, p=0.011) or CRC deaths (hazard ratios=0.426, p=0.039). However, no significant correlation is found between the methylation of MGMT promoter and the prognosis of CRC subjects. In addition, no interaction between 5-FU adjuvant chemotherapy and methylation of the two genes are observed. CONCLUSIONS: Methylation in the APC promoter may serve as a predictor for the prognosis of Taiwanese CRC patients. PMID- 19309277 TI - Detection of parental origin and cell stage errors of a double nondisjunction in a fetus by QF-PCR. AB - AIM: To investigate parental origins and cell stage errors of a double nondisjunction in a fetus. METHOD: For the determination of the most common chromosome anomalies, quantitative fluorescent polymerase chain reaction method using short tandem repeat (STR) DNA markers was applied to a fetus with abnormal ultrasonographic findings. Parental origin and cell stage errors of the trisomies were inferred by comparing the inherited STR alleles. Conventional cytogenetic technique was also applied for the confirmation of the aneuploidies. RESULTS: A double nondisjunction including chromosomes 21 and X (48,XXX,+21) was detected prenatally in the fetus. The origin of both chromosomes was maternal, and the errors were in meiosis I for 21 and meiosis II for X. Molecular results were concordant with cytogenetic results. CONCLUSION: Molecular techniques could be useful for the pre- and postnatal diagnosis of the common aneuploidies and determining its parental origin. This kind of study will improve knowledge about the mechanisms of nondisjunction and enable appropriate and rapid genetic counseling. PMID- 19309278 TI - 1q44-qter trisomy: clinical report and review of the literature. AB - Subtelomeric rearrangements are one of the main causes of multiple congenital anomalies and mental retardation, and they are detected in 5% of patients. We report on a 6.5-year-old boy with mental retardation, dysmorphic features, and behavioral problems, who revealed 1q44-qter trisomy and 22q13.3-qter monosomy due to a maternal cryptic translocation t(1;22). We compared the clinical and cytogenetic data of our patient with those of another case presenting a pure 22qter monosomy and with those of all 1qter trisomy cases reported in the international literature. To the best of our knowledge, the subterminal 1q trisomy found in the present case has been reported in only 12 patients to date (including five familial cases). This report aims to contribute to our understanding of 1q44-qter trisomy. PMID- 19309279 TI - Detection of MEFV gene mutations in patients with inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) with ulcerative colitis (UC) and Crohn's disease (CD) as the most common forms is an inflammation of the gastrointestinal tract. Familial Mediterranean fever (FMF) is another inflammatory disease as well. In the current study we studied FMF gene mutations in 47 patients with IBD and 25 healthy individuals to investigate the effects of these mutations on the clinical status of IBD. Twelve mutations were analyzed by reverse hybridization after multiplex PCR amplification of DNA samples. We did not find an association between FMF gene mutations and IBD phenotypic characteristics. However, in patients without Mediterranean fever (MEFV) mutations, extraintestinal disease frequencies were higher (p<0.05). IBD has a genetic basis with multiple genes probably playing a role via several pathways during disease progression. Studying other genes interacting with FMF gene in a larger group of patients will add to the knowledge of disease pathogenesis. PMID- 19309280 TI - Distribution of killer cell immunoglobulin-like receptor (KIR) genotypes in patients with familial Mediterranean fever. AB - Genotypic profiles of the natural killer cell immunoglobulin-like receptors (KIR) have been reported to vary among different ethnic groups and variable clinical entities. This study represents the first report on its distribution among patients with familial Mediterranean fever (FMF). We studied 56 unrelated Lebanese FMF patients, had their DNA typed using sequence-specific primer (SSP) technique for the presence of 16 KIR gene and pseudogene loci, and compared them to the general Lebanese population. The AA1 genotype was the most frequent in both the FMF and control groups. Six new KIR profiles were identified. The FMF group showed a higher prevalence of KIR 3DP1*003 (p<0.05) and an increase in the BB genotype compared with controls. The results lead to an interesting future research question of whether or not KIR genotype is involved in the predisposition to or pathogenesis of FMF. This is the first report that describes the KIR genotypic profile in this important clinical disease. PMID- 19309281 TI - High-throughput identification of mutations using a combination of CEL I fragmentation and SAGE technology. AB - A new method to detect mutations based on the serial analysis of gene expression (SAGE) technique, ligation-mediated (LM) PCR, and recombinant nuclease CEL I named LM-SAGE assay is reported in the present study. Mismatched DNA heteroduplexes formed from wild-type and mutant DNA are fragmented with CEL I nuclease at the mutant site to produce a double-strand fragment with an overhanging base at the 3'-end. The fragment is ligated to a linker, and digested with MmeI and then ligated to another linker. PCR is performed to amplify the ligation products, and NlaIII is used to release 17-bp tags containing mutation sites followed by purification, concatemerization, cloning, and sequencing. The locations of mutations can be identified from the homology analysis of tags. This new LM-SAGE assay can detect both known and unknown mutations with a sensitivity of 1:50 (mutant:wild-type DNA ratio) in 2.4 x 10(6) copies starting DNA sample. Our results show that this method could be used as a potentially high-throughput assay for mutation detection, particularly for the discovery of unknown mutations in genomic DNA. PMID- 19309282 TI - Dysferlinopathy in Chile: evidence of two novel mutations in the first reported cases. AB - We describe two Chilean patients with dysferlinopathy, a 32-year-old man with Miyoshi's distal myopathy and a 29-year-old woman with a proximodistal phenotype. Absence of dysferlin in frozen muscle biopsy allowed diagnostic confirmation. In these two patients, two mutations not previously identified in other populations were found: a homozygous c.1948delC (p.Leu650TyrfsX6) was found in the male patient; the heterozygous mutation c.1276G>A (p.Gly426Arg) was found in the female patient in association with the previously reported c.2858dupT (p.Phe954ValfsX2). To our knowledge, this is the first time that mutations in DYSF are identified in native Chileans. Our findings suggest the possibility that mutations in the DYSF gene were present in the Native American population before colonization. PMID- 19309283 TI - A novel MECP2 gene mutation in a Tunisian patient with Rett syndrome. AB - Patients with classical Rett show an apparently normal psychomotor development during the first 6-18 months of life. Thereafter, they enter a short period of developmental stagnation followed by a rapid regression in language and motor development. Purposeful hand use is often lost and replaced by repetitive, stereotypic movements. Rett syndrome (RTT) is an X-linked dominant disorder caused frequently by mutations in the methyl-CpG-binding protein 2 gene (MECP2). The aim of this study was to search for mutations in MECP2 gene in two Tunisian patients affected with RTT. The results of mutation analysis revealed mutations in exon 4 of MECP2 gene in the two patients. In one patient we identified a new mutation consisting of a deletion of four bases (c.810-813delAAAG), which led to a frame shift and generated a premature stop codon (p.Lys271Arg fs X15) in transcriptional repression domain-nuclear localization signal (TRD-NLS) domain of MeCP2 protein. With regard to the second patient, a previously described transition (c.916C>T) that changed an arginine to a cysteine residue (p.R306C) in TRD domain of MeCP2 protein was revealed. In conclusion, a new and a known de novo mutation in MECP2 gene were revealed in two Tunisian patients affected with RTT. PMID- 19309284 TI - Haplotype diversity and linkage disequilibrium at DRD2 locus--a study on four population groups of Andhra Pradesh, India. AB - Dopamine receptor D2 (DRD2) is expressed in the central nervous system and has a high affinity for many antipsychotic drugs. Besides several epidemiological investigations on association of DRD2 locus polymorphism(s) with neuropsychiatric problems and addictive behavior, a few polymorphisms in this locus have also been used to understand genomic diversity and population migratory histories globally. The present study attempts to understand the genomic diversity/affinity among four endogamous groups of Andhra Pradesh (India) against the backdrop of diversity studies from other parts of India and the rest of the world, with special reference to DRD2 locus. The four population groups from Adilabad District of Andhra Pradesh, namely, Brahmin (n=50), Nayakpod (n=49), Thoti (n=52), and Kolam (n=53), were included in the study. The DRD2 markers typed for the present study are three biallelic restriction fragments, that is, TaqI A (rs1800497), TaqI B (rs1079597), and TaqI D (rs1800498). Scoring of DRD2 haplotypes with respect to the three TaqI sites shows that five out of eight possible haplotypes are shared by the four populations. Ancestral haplotype B2D2A1 is most frequent among Thotis (0.359). The results of the present study indicate a differential gene flow into South India followed by certain important demographic events resulting in diversified peopling of India. PMID- 19309285 TI - Attitudes and knowledge about genetic testing before and after finding the disease-causing mutation among individuals at high risk for familial, early-onset Alzheimer's disease. AB - This study was designed to investigate the influence of finding a disease-causing mutation for an early-onset form of Alzheimer's disease, with the intention of taking a presymptomatic genetic test. First-degree relatives of patients with Alzheimer's disease from a large Cuban family with a newly described mutation in presenilin 1 gene were interviewed before and after molecular studies. Significant differences were observed regarding the knowledge of the disease (p=0.0004), interest in presymptomatic testing (p=0.000), and possible reproductive behavior (p=0.0087) in the same individuals in two different periods, 1997 and 2007. Our results show that there is a marked difference in the attitudes concerning genetic testing when individuals gain more knowledge about the disease and when there is more certainty about the test. PMID- 19309286 TI - Prenatal diagnosis of Cockayne syndrome type A based on the identification of two novel mutations in the ERCC8 gene. AB - Back Cockayne syndrome (CS; MIM 133540-216400) is a rare autosomal recessive neurodegenerative disorder characterized by progressive growth failure, microcephaly, mental retardation, retinal pigmentary degeneration, deafness, photosensitivity, accelerated systemic degeneration of somatic tissue, and premature death. Complementation assays have defined Cockayne syndrome group A (CSA) and Cockayne syndrome group B (CSB), caused by mutations in ERCC8 and ERCC6. The aim of this work was to perform a molecular analysis in a family with an affected son, who died at the age of 12, presenting clinical features typical of CSA. Molecular analysis of ERCC8 allowed us to characterize two novel mutations: a maternally inherited deletion encompassing exons 5 and 6, and a nonsense mutation located in exon 4, segregating from the father. Based on this molecular characterization, we successively performed a prenatal diagnosis on chorionic villus sampling, at 11th week of pregnancy. Molecular prenatal analysis of the ERCC8 was done by analyzing fetal DNA and RNA, looking for both mutations identified in the proband. A linkage analysis was performed using microsatellite markers located on chromosome 5q11 with the purpose to follow the segregation of the mutated alleles within the family. The fetal genotype at CSA locus resulted wild type and was confirmed at birth on biological material isolated from placenta. This study documents for the first time a molecular prenatal diagnosis of CSA, which results in the preferred approach if the mutation within the family is identified in a timely manner. PMID- 19309287 TI - Pregnancy as foreground in cystic fibrosis carrier testing decisions in primary care. AB - Cystic fibrosis carrier testing (CFCT) is among the first of the DNA tests offered prenatally in primary care settings. This paper from a descriptive qualitative study describes the influence of pregnancy in CFCT decisions by women receiving community-based prenatal care. Twenty-seven women receiving prenatal care in Midwestern U.S. primary care clinics completed semistructured interviews. Audiotaped interviews were analyzed using content analysis. Participants described decision-making influences and strategies from the perspective of "being pregnant." Patterns of attitudes and beliefs include (1) dealing with emotions, (2) pregnancy is natural, and (3) thinking about the baby. Strategies in the decision-making process included (1) reducing stress, (2) choosing what is relevant, (3) doing everything right, (4) wanting to be prepared, (5) delaying information, and (6) trusting God. While other factors were mentioned by some women, major themes reflect the influence of currently being pregnant on the decision-making process. These findings suggest that pregnancy is a powerful influence on the decision-making process and may not be the optimal time to make fully informed decisions regarding genetic carrier testing. Further understanding of factors influencing the genetic testing decision-making process is needed. Offering CFCT prior to conception is advocated. PMID- 19309288 TI - Gilbert syndrome as a predisposing factor for cholelithiasis risk in the Greek adult population. AB - We investigated the hypothesis that coinheritance of the common A(TA)(n)TAA promoter mutation at the UGT1A1 locus associated with Gilbert syndrome is a risk factor for gallstone formation in a homogeneous adult population, by conducting a case-control study that included 198 adult patients with cholelithiasis and 152 healthy controls both of Greek origin. Three genotypes were found: 7/7 (17.8% in controls and 23.3% in patients), 6/7 (33.5% in controls and 46.5% in patients), and normal homozygous 6/6 (48.7% in controls and 30.3% in patients). The Gilbert UGT1A1 genotypes 6/7 and 7/7 show significant association (odds ratio 2.225, 95% confidence interval 1.373-3.605, p=0.001, and odds ratio 2.101, 95% confidence interval 1.171-3.770, p=0.013, respectively) with cholelithiasis risk. This association supports the theory that genetic factors are responsible for a fraction of symptomatic gallstone disease; however, further studies are required in different ethnic groups to fully elucidate the involvement of Gilbert syndrome in gallstone disease. PMID- 19309289 TI - Screening of the DFNB3 locus: identification of three novel mutations of MYO15A associated with hearing loss and further suggestion for two distinctive genes on this locus. AB - Recessive mutations of MYO15A are associated with nonsyndromic hearing loss (HL) in humans (DFNB3) and in the shaker-2 mouse. Human MYO15A has 66 exons and encodes unconventional myosin XVA. Analysis of 77 Tunisian consanguineous families segregating recessive deafness revealed evidence of linkage to microsatellite markers for DFNB3 in four families. In two families, sequencing of MYO15A led to the identification of two novel homozygous mutations: a nonsense (c.4998C>A (p.C1666X) in exon 17 and a splice site mutation in intron 54 (c.9229 + 1G>A). A novel mutation of unknown significance, c.7395 + 3G>C, was identified in the third family, and no mutation was found in the fourth family. In conclusion, we discovered three novel mutations of MYO15A, and our data suggest the possibility that there are two distinct genes at the DFNB3 locus. PMID- 19309290 TI - Improved induction of immune tolerance to factor IX by hepatic AAV-8 gene transfer. AB - Gene therapy for hemophilia B has been shown to result in long-term expression and immune tolerance to factor IX (F.IX) after in vivo transduction of hepatocytes with adeno-associated viral (AAV-2) vectors in experimental animals. An optimized protocol was effective in several strains of mice with a factor 9 gene deletion (F9(-/-)). However, immune responses against F.IX were repeatedly observed in C3H/HeJ F9(-/-) mice. We sought to establish a gene transfer protocol that results in sustained expression without a requirement for additional manipulation of the immune system. Compared with AAV-2, AAV-8 was more efficient in transgene expression and induction of tolerance to F.IX in three different strains of wild-type mice. At equal vector doses, AAV-8 induced transgene product specific regulatory CD4(+)CD25(+)FoxP3(+) T cells at significantly higher frequency. Moreover, sustained correction of hemophilia B in C3H/HeJ F9(-/-) mice without antibody formation was documented in all animals treated with > or =4 x 10(11) vector genomes (VG)/kg and in 80% of mice treated with 8 x 10(10) VG/kg. Therefore, it is possible to develop a gene transfer protocol that reliably induces tolerance to F.IX largely independent of genetic factors. A comparison with other studies suggests that additional parameters besides plateau levels of F.IX expression contributed to the improved success rate of tolerance induction. PMID- 19309291 TI - Brain structures involved in visual search in the presence and absence of color singletons. AB - It is still debated to what degree top-down and bottom-up driven attentional control processes are subserved by shared or by separate mechanisms. Interactions between these attentional control forms were investigated using a rapid event related fMRI design, using an attentional search task. Following a prestimulus mask, target stimuli (consisting of a letter C or a mirror image of the C, enclosed in a diamond outline) were presented either at one unique location among three nontarget items (consisting of a random letter, enclosed in a circle outline; 50% probability), or at all four possible target locations (also 50% probability). On half the trials, irrelevant color singletons were presented, consisting of a color change of one of the four prestimulus masks, just prior to target appearance. Participants were required to search for a target letter inside the diamond and report its orientation. Results indicate that, in addition to a common network of parietal areas, medial frontal cortex is uniquely involved in top-down orienting, whereas bottom-up control is mainly subserved by a network of occipital and parietal areas. Additionally, we found that participants who were better able to suppress orienting to the color singleton showed middle frontal gyrus activation, and that the degree of top-down control correlated with insular activity. We conclude that, in addition to a common set of parietal areas, separate brain areas are involved in top-down and bottom-up driven attentional control, and that frontal areas play a role in the suppression of attentional capture by an irrelevant color singleton. PMID- 19309292 TI - On the temporal relation of top-down and bottom-up mechanisms during guidance of attention. AB - Two mechanisms are said to be responsible for guiding focal attention in visual selection: bottom-up, saliency-driven capture and top-down control. These mechanisms were examined with a paradigm that combined a visual search task with postdisplay probe detection. Two SOAs between the search display and probe onsets were introduced to investigate how attention was allocated to particular items at different points in time. The dynamic interplay between bottom-up and top-down mechanisms was investigated with ERP methodology. ERPs locked to the search displays showed that top-down control needed time to develop. N2pc indicated allocation of attention to the target item and not to the irrelevant singleton. ERPs locked to probes revealed modulations in the P1 component reflecting top down control of focal attention at the long SOA. Early bottom-up effects were observed in the error rates at the short SOA. Taken together, the present results show that the top-down mechanism takes time to guide focal attention to the relevant target item and that it is potent enough to limit bottom-up attentional capture. PMID- 19309293 TI - Neural correlates of human body perception. AB - The objective of this study was to investigate potential sex differences in the neural response to human bodies using fMRI carried out in healthy young adults. We presented human bodies in a block-design experiment to identify body responsive regions of the brain, namely, extrastriate body area (EBA) and fusiform body area (FBA). In a separate event-related "adaptation" experiment, carried out in the same group of subjects, we presented sets of four human bodies of varying body size and shape. Varying levels of body morphing were introduced to assess the degree of morphing required for adaptation release. Analysis of BOLD signal in the block-design experiment revealed significant Sex x Hemisphere interactions in the EBA and the FBA responses to human bodies. Only women showed greater BOLD response to bodies in the right hemisphere compared with the left hemisphere for both EBA and FBA. The BOLD response in right EBA was higher in women compared with men. In the adaptation experiment, greater right versus left hemisphere response for EBA and FBA was also identified among women but not men. These findings are particularly novel in that they address potential sex differences in the lateralization of EBA and FBA responses to human body images. Although previous studies have found some degree of right hemisphere dominance in body perception, our results suggest that such a functional lateralization may differ between men and women. PMID- 19309294 TI - Effects of 10 Hz rTMS on the neural efficiency of working memory. AB - Working memory (WM) has been described as short-term retention of information that is no longer accessible in the environment, and the manipulation of this information for subsequent use in guiding behavior. WM is viewed as a cognitive process underlying higher-order cognitive functions. Evidence supports a critical role for PFC in mediating WM performance. Studies show psychomotor processing speed and accuracy account for considerable variance in neural efficiency (Ne). This study compared the relative effects of active and sham 10 Hz rTMS applied to dorsolateral prefrontal cortex (DLPFC) on indices of Ne in healthy participants performing a WM paradigm that models the association between WM load and task behavior [Sternberg, S. High-speed scanning in human memory. Science, 153, 652 654, 1966]. Previous studies identified a relationship between diminished Ne and impaired WM across a broad array of clinical disorders. In the present study, the authors predicted there would be a main effect of stimulation group (STM) on accuracy (SCR) and processing speed (RT), hence, Ne. We observed a main effect of STM for RT without an effect on SCR; even so, there was a robust effect of STM on Ne. PMID- 19309295 TI - Monitoring coordination during bimanual movements: where is the mastermind? AB - One remarkable aspect of the human motor repertoire is the multitude of bimanual actions it contains. Still, the neural correlates of coordinated movements, in which the two hands share a common goal, remain debated. To address this issue, we designed two bimanual circling tasks that differed only in terms of goal conceptualization: a "coordination" task that required movements of both hands to adapt to each other to reach a common goal and an "independent" task that imposed a separate goal to each hand. fMRI allowed us to pinpoint three areas located in the right hemisphere that were more strongly activated in the coordination condition: the superior temporal gyrus (STG), the SMA, and the primary motor cortex (M1). We then used transcranial magnetic stimulation (TMS) to disrupt transiently the function of those three regions to determine their causal role in bimanual coordination. Right STG virtual lesions impaired bimanual coordination, whereas TMS to right M1 enhanced hand independence. TMS over SMA, left STG, or left M1 had no effect. The present study provides direct insight into the neural correlates of coordinated bimanual movements and highlights the role of right STG in such bimanual movements. PMID- 19309296 TI - Blindsight mediated by an S-cone-independent collicular pathway: an fMRI study in hemispherectomized subjects. AB - The purpose of our study was to investigate the ability to process achromatic and short-wavelength-sensitive cone (S-cone)-isolating (blue-yellow) stimuli in the blind visual field of hemispherectomized subjects and to demonstrate that blindsight is mediated by a collicular pathway that is independent of S-cone inputs. Blindsight has been described as the ability to respond to visual stimuli in the blind visual field without conscious awareness [Weiskrantz, L., Warrington, E. K., Sanders, M. D., & Marshall, J. Visual capacity in the hemianopic field following a restricted occipital ablation. Brain, 97, 709-728, 1974]. The roles of the subcortical neural structures in blindsight, such as the pulvinar and the superior colliculus, have been debated and an underlying neural correlate has yet to be confirmed. Using fMRI, we tested the ability to process visual stimuli that isolated the achromatic and short-wavelength-sensitive (S-) cone pathways in three subjects: one control subject, one hemispherectomized subject with blindsight, and one hemispherectomized subject without blindsight. We demonstrated that (1) achromatic and S-cone-isolating stimuli presented to the normal visual hemifield of hemispherectomized subjects and to both visual hemifields of the control subject activated contralateral visual areas (V1/V2), as expected; (2) achromatic stimulus presentation but not S-cone-isolating stimulus presentation to the blind hemifield of the subject with blindsight activated visual areas FEF/V5; (3) whereas the cortical activation of the control subject was enhanced by an additional stimulus (achromatic and S-cone isolating) presented in the contralateral visual field, activation pattern of the subject with blindsight was enhanced by achromatic stimuli only. We conclude that the human superior colliculus is blind to the S-cone-isolating stimuli, and blindsight is mediated by an S-cone-independent collicular pathway. PMID- 19309297 TI - Higher milk requirements for bone mineral accrual in adolescent girls bearing specific caucasian genotypes in the VDR promoter. AB - Low milk intakes hamper bone mineral acquisition during adolescence, especially in European girls. We hypothesized that ethnic-specific polymorphisms of the vitamin D receptor gene promoter (VDRp) influence this milk/bone association. We evaluated lumbar spine BMC and BMD, milk/dairy products and calcium intakes, markers of P-Ca metabolism, and VDRp polymorphisms at the Cdx-2 binding (rs11568820) and -1012 (rs4516035) loci in 117 healthy European peri- and postmenarcheal girls (14.9 +/- 1.6 yr) during a 4-yr follow-up. Calcium intakes from milk, nonmilk dairy products, and nondairy products averaged 199, 243, and 443 mg/d at the initiation of the study. Results show no association between milk intakes and bone mass accrual in girls bearing an A/A genotype at the -1012 VDRp locus (30% of the cohort). In contrast, A/G or G/G girls had lower spine BMC ( 13%, p = 0.031), BMD (-10%, p = 0.004), and BMD Z-score (-0.84 SD, p = 0.0003) when their milk intakes were <260 ml/d compared with genotype-matched girls with higher milk intakes and with girls with an A/A genotype. The negative impact of low milk intake persisted up to 19.0 +/- 1.7 yr. These findings suggest that European girls bearing a -1012 A/G or G/G VDRp genotype should have higher milk/calcium intakes for optimal vertebral mass accrual during adolescence than girls bearing an A/A genotype, a genotype found in 30% of European and 98% of Asian and Sub-Saharan African populations. VDRp genotype diversity may contribute to the ethnic differences observed in milk requirements for bone health during adolescence. PMID- 19309298 TI - Dairy versus calcium carbonate in promoting peak bone mass and bone maintenance during subsequent calcium deficiency. AB - Dairy products provide most of the calcium in the diet, but consumption of milk has declined steadily over the last six decades. We determined (1) benefits to bone formation in growing female Sprague-Dawley rats when calcium is derived from dairy versus CaCO(3) and (2) residual benefits of calcium provided by dairy to rats subsequently receiving a low-calcium diet. During growth, femurs from rats fed nonfat dry milk solids (NFDM) had 8.4% higher peak breaking force, 6.4% greater Ca content, 4.8% greater weight, 4% greater width, 1.2% greater density, 13.1% greater midshaft cortical thickness, and 16.7% greater midshaft cortical area than from rats fed CaCO(3). These effects were unrelated to differences in calcium absorption or serum IGF-1, but the NFDM group had higher rates of bone formation. If maintained on an adequate calcium diet, many of these advantages disappeared. However, rats fed adequate Ca as NFDM versus CaCO(3) during growth and subsequently switched to deficient Ca as CaCO(3) had significantly (p < 0.0001) higher femoral BMD (1.3%), total bone Ca (7.2%), Ca concentration (4.6%), and cortical thickness (9.4%) and a trend (p = 0.02) toward greater peak breaking force (17%). Thus, NFDMs improved bone measures during growth and protected bone against a subsequent period of calcium depletion compared with CaCO(3). PMID- 19309299 TI - Sporadic and MEN1-related primary hyperparathyroidism: differences in clinical expression and severity. AB - Primary hyperparathyroidism (PHPT) is a common endocrine disease that is associated with multiple endocrine neoplasia type 1 (MEN1) in approximately 2% of PHPT cases. Lack of a family history and other specific expressions may lead to underestimated MEN1 prevalence in PHPT. The aim of this study was to identify clinical or biochemical features predictive of MEN1 and to compare the severity of the disease in MEN1-related versus sporadic PHPT (sPHPT). We performed a 36-mo cross-sectional observational study in three tertiary referral centers on an outpatient basis on 469 consecutive patients with sporadic PHPT and 64 with MEN1 related PHPT. Serum calcium, phosphate, PTH, 25(OH)D(3), and creatinine clearance were measured, and ultrasound examination of the urinary tract/urography was performed in all patients. In 432 patients, BMD was measured at the lumbar spine (LS) and femoral neck (FN). MEN1 patients showed lower BMD Z-scores at the LS ( 1.33 +/- 1.23 versus -0.74 +/- 1.4, p = 0.008) and FN (-1.13 +/- 0.96 versus -0.6 +/- 1.07, p = 0.002) and lower phosphate (2.38 +/- 0.52 versus 2.56 +/- 0.45 mg/dl, p = 0.003) and PTH (113.8 +/- 69.5 versus 173.7 +/- 135 pg/ml, p = 0.001) levels than sPHPT patients. Considering probands only, the presence of MEN1 was more frequently associated with PTH values in the normal range (OR, 3.01; 95% CI, 1.07-8.50; p = 0.037) and younger age (OR, 1.61; 95% CI, 1.28-2.02; p = 0.0001). A combination of PTH values in the normal range plus age <50 yr was strongly associated with MEN1 presence (OR, 13.51; 95% CI, 3.62-50.00; p = 0.0001). In conclusion, MEN1-related PHPT patients show more severe bone but similar kidney involvement despite a milder biochemical presentation compared with their sPHPT counterparts. Normal PTH levels and young age are associated with MEN1 presence. PMID- 19309300 TI - Erythropoietin-induced optimization of renal function after warm ischemia. AB - BACKGROUND AND PURPOSE: Recent preclinical data have indicated that erythropoietin (Epo) can protect organs from ischemic damage. We evaluated the ability of Epo to protect the kidney from the effects of ischemia. METHODS: Thirty dogs underwent a laparoscopic nephrectomy and were allowed to recover for 2 weeks. The dogs were then divided into five groups. Animals in groups 1 and 2 underwent 1.5 hours of abdominal insufflation with placebo (saline) injection (group 1) or Epo injection (group 2) before; groups 3 to 5 underwent 1 hour of laparoscopic renal artery clamping after placebo injection (group 3), Epo injection (group 4), or mannitol injection (group 5). Serum evaluations and 24 hour urine collections were performed weekly. After 28 days, the animals were sacrificed. Statistical analysis was performed with the Kruskal-Wallis test. RESULTS: After recovery from the initial nephrectomy, all dogs had similar serum hematocrit and creatinine levels. Hematocrit was not significantly affected by Epo administration at any time point. Immediately after the second surgery, dogs that underwent renal artery clamping (groups 3-5) had significantly lower 24-hour urine creatinine levels than those that were not clamped (groups 1-2). After 4 weeks of recovery, the dogs that had received Epo before ischemia (group 4) had recovered significantly more renal function than the dogs that received placebo or mannitol before ischemia (urine creatinine level = Epo 149.1 mg/dL v placebo 70.7 mg/dL v mannitol 80.7 mg/dL). At sacrifice, microalbuminuria was also significantly less in dogs receiving Epo before ischemia than their mannitol or placebo counterparts. CONCLUSION: The current study demonstrates that administering Epo before warm ischemia can improve the recovery of renal function after ischemia better than placebo or mannitol. PMID- 19309301 TI - Ureteroscopy in pregnant women for ureteral stone. AB - INTRODUCTION: The occurrence of urolithiasis in pregnancy represents a challenge in both diagnosis and treatment of this condition, because it presents risks not only to the mother but also to the fetus. Surgical treatment may be indicated for patients with infection, persistent pain, and obstruction of a solitary kidney. We present our experience on the management of pregnant patients with ureteral calculi and a review of the literature. MATERIALS AND METHODS: The charts of 19 pregnant patients with obstructive ureteral calculi were retrospectively reviewed. Gestational age ranged from 13 to 33 weeks. In all patients, ureteral stone was diagnosed on abdominal ultrasound. In regard to localization, 15 calculi were in the distal ureter, 3 in the proximal ureter, and 1 in the interior of an ureterocele. Calculi size ranged from 6 to 10 mm (mean, 8 mm). The following criteria were used to indicate ureteroscopy: persistent pain with no improvement after clinical treatment, increase in renal dilation, or presence of uterine contractions. Nine patients (47.3%) were submitted to ureteroscopy. All calculi (100%) were removed with a stone basket extractor under continuous endoscopic vision. None of the calculi demanded the use of a lithotriptor. RESULTS: Nine patients (47.3%) treated with clinical measurements presented no obstetric complications and spontaneous elimination of the calculi. Nine patients (47.3%) submitted to ureteroscopy had no surgical complications. There was remission of pain in all cases after ureteroscopy and ureteral catheter placement. CONCLUSION: The diagnosis and treatment of ureteral lithiasis in pregnant women present potential risks for the fetus and the mother. Conservative management is the first option, but ureteroscopy may be performed with safety and high success rates. PMID- 19309305 TI - Has human immunodeficiency virus become more virulent? PMID- 19309306 TI - Is HIV becoming more virulent? Initial CD4 cell counts among HIV seroconverters during the course of the HIV epidemic: 1985-2007. AB - BACKGROUND: Whether human immunodeficiency virus (HIV) seroconverters have been presenting with progressively lower CD4 cell counts over the course of the HIV epidemic is controversial. Additional data on whether HIV might have become more virulent on a population level (measured by post-seroconversion CD4 cell counts) may provide important insights regarding HIV pathogenesis. METHODS: To determine whether post-seroconversion CD4 cell counts have changed over time, we evaluated 2174 HIV seroconverters as part of a large cohort study during the period 1985 2007. Participants were documented antiretroviral-naive HIV seroconverters who had a CD4 cell count measured within 6 months after receiving a diagnosis of HIV infection. Multiple linear regression models were used to assess trends in initial CD4 cell counts. RESULTS: The mean initial CD4 cell count decreased during the study period from 632 cells/mm(3) in 1985-1990 to 553 cells/mm(3) in 1991-1995, 493 cells/mm(3) in 1996-2001, and 514 cells/mm(3) in 2002-2007. During those periods, the percentages of seroconverters with an initial CD4 cell count <350 cells/mm(3) were 12%, 21%, 26%, and 25%, respectively. In the multiple linear model, the mean decrease in CD4 cell count from 1985-1990 was 65 cells/mm(3) in 1991-1995 (P < .001)), 107 cells/mm(3) in 1996-2001 (P < .001), and 102 cells/mm(3) in 2002-2007 (P < .001). Similar trends occurred with regard to CD4 cell percentage and total lymphocyte count. Similar decreases in initial CD4 cell counts were observed among African American and white persons during the epidemic. DISCUSSION: A significant decrease in initial CD4 cell counts among HIV seroconverters in the United States has occurred during the HIV epidemic. These data provide an important clinical correlate to suggestions that HIV may have adapted to the host, resulting in a more virulent infection. PMID- 19309307 TI - Is the interruption of antiretroviral treatment during pregnancy an additional major risk factor for mother-to-child transmission of HIV type 1? AB - BACKGROUND: There is currently an experts' agreement discouraging interruption of antiretroviral treatment (ART) during the first trimester of pregnancy in women infected with human immunodeficiency virus type 1 (HIV-1). However, this recommendation is poorly supported by data. We evaluated the effects of discontinuing ART during pregnancy on the rate of mother-to-child transmission. METHODS: Logistic regression models were performed in a prospective cohort of 937 children who were perinatally exposed to HIV-1 to estimate adjusted odds ratios for confounding factors on mother-to-child transmission, including maternal interruption of ART. RESULTS: Among 937 pregnant women infected with HIV-1, ART was interrupted in 81 (8.6%) in the first trimester and in 11 (1.2%) in the third trimester. In the first trimester, the median time at suspension of ART was 6 weeks (interquartile range [IQR], 5-6 weeks) and the time without treatment was 8 weeks (IQR, 7-11 weeks). In the third trimester, the median time at suspension of ART was 32 weeks (IQR, 23-36 weeks) and the time without treatment was 6 weeks (IQR, 2-9 weeks). The plasma viral load was similar in women who had treatment interrupted in the first trimester and in those who did not have treatment interrupted. Overall, the rate of mother-to-child transmission in the whole cohort was 1.3% (95% confidence interval [CI], 0.7%-2.3%), whereas it was 4.9% (95% CI, 1.9%-13.2%) when ART was interrupted in the first trimester and 18.2% (95% CI, 4.5%-72.7%) when ART was interrupted in the third trimester. In the multiple logistic regression models, only interruption of ART during either the first or the third trimester, maternal mono- or double therapy, delivery by a mode other than elective cesarean delivery, and a viral load at delivery >4.78 log(10) copies/mL were independently associated with an increased rate of mother to-child transmission. CONCLUSIONS: Discontinuing ART during pregnancy increases the rate of mother-to-child transmission of HIV-1, either when ART is stopped in the first trimester and subsequently restarted or when it is interrupted in the third trimester. This finding supports recommendations to continue ART in pregnant women who are already receiving treatment for their health. PMID- 19309308 TI - Association of interleukin-18 gene promoter polymorphisms with risk of acute myocardial infarction in northern Chinese Han population. AB - BACKGROUND: Interleukin-18 (IL-18) has been suggested to play an important role in coronary arterial disease and its sequelae. The aim of the present study was to investigate the association between IL-18 promoter functional polymorphisms ( 607C/A and -137G/C) and acute myocardial infarction (AMI) in northern Chinese Han population. METHODS: We performed a case-control study including 234 patients with AMI and 216 age- and sex-matched controls. Genotyping was performed using sequence specific primer-polymerase chain reaction (PCR-SSP). RESULTS: There were significant differences in the genotype and allele distribution of -607C/A polymorphism of the IL-18 gene between cases and controls. Logistic regression analysis with adjustments for other well-established risk factors revealed that the -607C allele carriers had a significantly increased risk of AMI compared with the non-carriers (OR=1.890, 95% CI 1.196-2.985, p=0.006). No relationship between -137G/C polymorphism and AMI was found. CONCLUSIONS: This study shows for the first time that the IL-18 gene promoter -607C/A polymorphism may be considered a genetic risk factor for AMI in northern Chinese Han population. PMID- 19309309 TI - RNA helicase A acts as a bridging factor linking nuclear beta-actin with RNA polymerase II. AB - Actin, the major component of the cytoplasmic skeleton, has been shown to exist in the nucleus. Nuclear actin functions in several steps of the transcription process, including chromatin remodelling and transcription initiation and elongation. However, as a part of PICs (pre-initiation complexes), the role of actin remains to be elucidated. In the present study, we identified RHA (RNA helicase A) as an actin-interacting protein in PICs. Using immunoprecipitation and immunofluorescence techniques, we have shown that RHA associates with beta actin in the nucleus. A GST (glutathione transferase) pulldown assay using different deletion mutants revealed that the RGG (Arg-Gly-Gly) region of RHA was responsible for the interaction with beta-actin, and this dominant-negative mutant reduced the recruitment of Pol II (RNA polymerase II) into PICs. Moreover, overexpression or depletion of RHA could influence the interaction of Pol II with beta-actin and beta-actin-involved gene transcription regulation. These results suggest that RHA acts as a bridging factor linking nuclear beta-actin with Pol II. PMID- 19309310 TI - Triton X-100 promotes a cholesterol-dependent condensation of the plasma membrane. AB - The molecular components of membrane rafts are frequently defined by their biochemical partitioning into detergent-resistant membranes. In the present study, we used a combination of epifluorescence and two-photon microscopy to visualize and quantify whether this insolubility in detergent reflects a pre existing organization of the PM (plasma membrane). We found that the treatment of cells with cold TX (Triton X-100) promotes a profound remodelling of the PM, including a rapid rearrangement of the glycosphingolipid GM1 and cholesterol into newly formed structures, only partial solubilization of fluid domains and the formation of condensed domains that cover 51% of the remaining membrane. TX does not appear to induce the coalescence of pre-existing domains; instead, the domains that remain after TX treatment seem to be newly formed with a higher degree of condensation than those observed in native membranes. However, when cholesterol was complexed physically by treatment with a second detergent, such as saponin, cholesterol did not separate into the newly formed structures, condensation of the domains was unaltered, and the relative area corresponding to ordered domains increased to occupy 62% of the remaining membrane. Our results suggest that detergent can be used to enrich ordered domains for biochemical analysis, but that TX treatment alone substantially alters the lateral organization of the PM. PMID- 19309311 TI - Drug screening by crossing membranes: a novel approach to identification of trypanocides. AB - Trypanosomes are a group of protozoan parasites that inflict huge health and economic burdens across the globe. The African trypanosome, Trypanosoma brucei, the causative agent of sleeping sickness, has a highly sophisticated mechanism of antigenic variation that facilitates chronic survival in the mammalian host, and also all but eliminates any realistic hope for vaccination-based control. However, trypanosomes are also highly divergent organisms, with many biochemical processes setting them apart from their hosts, and there remains great optimism that these features may be exploited for development of new drugs. Unfortunately, the compounds that are in use at present are decades old and resistance has emerged. The article in this issue of the Biochemical Journal by Patham et al., a joint team from the universities of Pittsburgh and Georgia, represents one approach to exploiting this divergence. The authors of the study have exploited novel aspects of the biochemistry within the system for translocation of nascent polypeptides across the endoplasmic reticulum membrane to identify three compounds that are able to inhibit the process. They then demonstrate that these same compounds are both trypanocidal, but well tolerated by human tissue culture cells. These observations may present interesting new leads in the fight against trypanosomiasis, and potentially identify a new target that can be explored for therapeutic potential. PMID- 19309312 TI - Snf1-related protein kinases (SnRKs) act within an intricate network that links metabolic and stress signalling in plants. AB - The phosphorylation and dephosphorylation of proteins, catalysed by protein kinases and phosphatases, is the major mechanism for the transduction of intracellular signals in eukaryotic organisms. Signalling pathways often comprise multiple phosphorylation/dephosphorylation steps and a long-standing hypothesis to explain this phenomenon is that of the protein kinase cascade, in which a signal is amplified as it is passed from one step in a pathway to the next. This review represents a re-evaluation of this hypothesis, using the signalling network in which the SnRKs [Snf1 (sucrose non-fermenting-1)-related protein kinases] function as an example, but drawing also on the related signalling systems involving Snf1 itself in fungi and AMPK (AMP-activated protein kinase) in animals. In plants, the SnRK family comprises not only SnRK1, but also two other subfamilies, SnRK2 and SnRK3, with a total of 38 members in the model plant Arabidopsis. This may have occurred to enable linking of metabolic and stress signalling. It is concluded that signalling pathways comprise multiple levels not to allow for signal amplification, but to enable linking between pathways to form networks in which key protein kinases, phosphatases and target transcription factors represent hubs on/from which multiple pathways converge and emerge. PMID- 19309313 TI - Connexin43 phosphorylation: structural changes and biological effects. AB - Vertebrate gap junctions, composed of proteins from the connexin gene family, play critical roles in embryonic development, co-ordinated contraction of excitable cells, tissue homoeostasis, normal cell growth and differentiation. Phosphorylation of connexin43, the most abundant and ubiquitously expressed connexin, has been implicated in the regulation of gap junctional communication at several stages of the connexin 'life cycle', including hemichannel oligomerization, export of the protein to the plasma membrane, hemichannel activity, gap junction assembly, gap junction channel gating and connexin degradation. Consistent with a short (1-5 h) protein half-life, connexin43 phosphorylation is dynamic and changes in response to activation of many different kinases. The present review assesses our current understanding of the effects of phosphorylation on connexin43 structure and function that in turn regulate gap junction biology, with an emphasis on events occurring in heart and skin. PMID- 19309315 TI - Temperature, not LuxS, mediates AI-2 formation in hydrothermal habitats. AB - Quorum sensing provides the basis for coordinating community-wide, microbial behaviors in many mesophilic bacteria. However, little attention has been directed toward the possibility that such phenomena occur in extremely thermal microbial environments. Despite the absence of luxS in hyperthermophile genomes, autoinducer-2 (AI-2), a boronated furanone and proposed 'universal' interspecies mesophilic bacterial communication signal, could be formed by Thermotoga maritima and Pyrococcus furiosus through a combination of biotic and abiotic reaction steps. AI-2 did not, however, induce any detectable quorum-sensing phenotypes in these organisms, although transcriptome-based evidence of an AI-2-induced stress response was observed in T. maritima. The significance, if any, of AI-2 in hydrothermal habitats is not yet clear. Nevertheless, these results show the importance of considering environmental factors, in this case high temperatures, as abiotic causative agents of biochemical and microbiological phenomena. PMID- 19309314 TI - IscA/SufA paralogues are required for the [4Fe-4S] cluster assembly in enzymes of multiple physiological pathways in Escherichia coli under aerobic growth conditions. AB - IscA/SufA paralogues are the members of the iron-sulfur cluster assembly machinery in Escherichia coli. Whereas deletion of either IscA or SufA has only a mild effect on cell growth, deletion of both IscA and SufA results in a null growth phenotype in minimal medium under aerobic growth conditions. Here we report that cell growth of the iscA/sufA double mutant (E. coli strain in which both iscA and sufA had been in-frame-deleted) can be partially restored by supplementing with BCAAs (branched-chain amino acids) and thiamin. We further demonstrate that deletion of IscA/SufA paralogues blocks the [4Fe-4S] cluster assembly in IlvD (dihydroxyacid dehydratase) of the BCAA biosynthetic pathway in E. coli cells under aerobic conditions and that addition of the iron-bound IscA/SufA efficiently promotes the [4Fe-4S] cluster assembly in IlvD and restores the enzyme activity in vitro, suggesting that IscA/SufA may act as an iron donor for the [4Fe-4S] cluster assembly under aerobic conditions. Additional studies reveal that IscA/SufA are also required for the [4Fe-4S] cluster assembly in enzyme ThiC of the thiamin-biosynthetic pathway, aconitase B of the citrate acid cycle and endonuclease III of the DNA-base-excision-repair pathway in E. coli under aerobic conditions. Nevertheless, deletion of IscA/SufA does not significantly affect the [2Fe-2S] cluster assembly in the redox transcription factor SoxR, ferredoxin and the siderophore-iron reductase FhuF. The results suggest that the biogenesis of the [4Fe-4S] clusters and the [2Fe-2S] clusters may have distinct pathways and that IscA/SufA paralogues are essential for the [4Fe-4S] cluster assembly, but are dispensable for the [2Fe-2S] cluster assembly in E. coli under aerobic conditions. PMID- 19309316 TI - Ethanol enhances GABAB-mediated inhibitory postsynaptic transmission on rat midbrain dopaminergic neurons by facilitating GIRK currents. AB - It is largely accepted that an activation of the dopaminergic system underlies the recreational and convivial effects of ethanol. However, the mechanisms of action of this drug on the dopaminergic neurons are not fully understood yet. In the present study, we have used intracellular electrophysiological techniques (current and single-electrode voltage-clamp) to investigate the actions of ethanol on the gamma-aminobutyric acid (GABA)(B)-mediated inhibitory postsynaptic potentials (IPSPs) in rat midbrain dopaminergic neurons. Ethanol (10-200 mM) augmented, in a concentration-dependent and reversible manner, the amplitude of the GABA(B)-IPSP. In addition, the GABA(B) agonist baclofen generated G-protein gated inward rectifying K(+) channels (GIRK)-related membrane hyperpolarizations/outward currents that were potentiated by ethanol. The potentiating effect of ethanol persisted in tetrodotoxin (TTX)-treated neurons, suggesting a postsynaptic site of action. These effects of ethanol were not changed by manipulating adenyl cyclase, protein kinases and phospholipase C activity, or by chelating intracellular Ca(2+) with EGTA. Interestingly, the outward current caused by the intracytoplasmatic diffusion of the irreversible G protein activator GTPgammaS was transiently enhanced by ethanol. Our observations suggest that the action of ethanol occurs on activated GIRK channels downstream of the GABA(B) receptors. These enhancing effects of ethanol on GABA(B)-induced synaptic responses could modulate alcohol intake and the altered mental and motor performance of individuals in an acute intoxicative phase. PMID- 19309317 TI - Dynamics of free intracellular Ca2+ during synaptic and spike activity of cricket tibial motoneurons. AB - For all nervous systems, motoneurons are the main output pathway. They are involved in generating episodic motor activity as well as enduring motor rhythms. To determine whether changes in cytosolic Ca(2+) correlate with motor performance, we studied the spatiotemporal dynamics, mode of entry and role of free intracellular Ca(2+) in cricket (Gryllus bimaculatus) front leg tibial extensor and flexor motoneurons. Synaptic activation or intracellular depolarising current injection uniformly increased Ca(2+) with the same dynamics throughout the primary and secondary branches of the dendritic tree of all motoneurons. Ca(2+) rise times (mean tau(rise), 233-295 ms) were lower than decay times (mean tau(decay), 1927-1965 ms), and resulted in a Ca(2+) plateau during repetitive activation, such as during walking. The neurons therefore operate with a different Ca(2+) level during walking than during episodic leg movements. Ca(2+) enters the dendritic processes of motoneurons via a voltage-activated mechanism. Entry is driven by subthreshold excitation, and is largely independent of the neurons' spiking activity. To what extent ligand-activated mechanisms of Ca(2+) entry operate remains uncertain. We found no evidence for any prominent Ca(2+)-activated secondary currents in these motoneurons. Excitatory postsynaptic potentials evoked by extracellular stimulation of descending neurons were unaffected by the level of free intracellular Ca(2+). The activity of tibial motoneurons therefore appears to be only weakly dependent on the level of free intracellular Ca(2+) in dendrites. This is different to what has been found for many other neurons studied, and may represent an essential prerequisite for insect motoneurons to support a wide range of both episodic and rhythmic motor sequences underlying behaviour. PMID- 19309318 TI - The natriuretic effect of oxytocin blocks medial tuberomammillary polydipsia and polyuria in male rats. AB - Lesions of the tuberomammillary complex, a neuroanatomical system closely related to the hypothalamic supraoptic and paraventricular nuclei, induce strong polydipsia in male rats. It was recently demonstrated that this increase in water intake is immediate, persistent, follows circadian rhythms and appears to be related to sodium regulation. The present study found that urine osmolality was significantly lower in tuberomammillary-lesioned animals vs. their respective controls at 8:00 h after surgery. Therefore, the aim of the present study was to examine the natriuretic effect of intraperitoneal oxytocin (OT) administration on medial ventral tuberomammillary nucleus (E3) polydipsia and polyuria of lesioned and control male rats. At 24:00 h post-lesion, OT blocked the hyperdipsic and polyuric responses of E3-lesioned animals but not those of non-lesioned controls, which did however significantly increase their water intake. Moreover, urinary osmolality and sodium excretion increased in E3 -lesioned animals that received OT but not in lesioned controls receiving physiological saline (992 +/- 187.19 vs. 215.83 +/- 23.39 mOsm/kg; 1.68 +/- 0.13 vs. 0.47 +/- 0.1 mEq/L). At 48:00 h post-lesion, OT administration also induced a higher intake of water and of simultaneously offered hypertonic NaCl (1.5%) in E3-lesioned animals. These results are interpreted in terms of the hypothalamic systems involved in sodium and water homeostasis. PMID- 19309319 TI - Upper limb movement interruptions are correlated to freezing of gait in Parkinson's disease. AB - Freezing of gait (FOG) in patients with Parkinson's disease (PD) is a common problem of unknown origin, which possibly reflects a general motor control deficit. We investigated the relationship between the frequency of freezing episodes during gait and during a bimanual task in control and subjects with PD with and without FOG. Group differences in spatiotemporal characteristics were also examined as well as the effects of visual cueing. Twenty patients with PD in the off-phase of the medication cycle and five age-matched controls performed a repetitive drawing task in an anti-phase pattern on a digitizer tablet. The task was offered at two different speeds (comfortable and maximal) and two different amplitudes (small and large) with and without visual cueing. The results showed that freezing episodes in the upper limbs occurred in only 10.4% of patient trials and that their occurrence was correlated with FOG scores (Spearman's rho = 0.64). Overall, few spatiotemporal differences were found between freezers, non freezers and controls, except for an overshooting of the target amplitude in controls. Effects of visual cueing were largely similar in all groups, except for the variability of relative phase, which decreased in non-freezers and controls, and was unaffected in freezers. Despite the fact that general motor differences between subgroups were small, freezing episodes were manifest during a bimanual repetitive upper limb task and were correlated to FOG. Further study into upper limb movement breakdown is warranted to understand the parallel deficits that lead up to FOG. PMID- 19309320 TI - Differential representation of Pavlovian-instrumental transfer by prefrontal cortex subregions and striatum. AB - Environmental cues that once predicted reward can restore extinguished behavior directed toward that reward. This process may be modeled by the Pavlovian instrumental transfer (PIT) paradigm where a previously learned Pavlovian conditioned stimulus (CS) elicits a representation of the reward associated with that CS, prompts motivation toward the absent reward, and triggers an instrumental action. We recorded in the medial and orbital prefrontal cortex (mPFC and OFC) and dorsal striatum (DS) of freely moving rats during PIT and found that a Pavlovian CS, as compared with neutral or no stimuli, amplified the phasic neuronal responses to instrumental nosepokes ('transfer' event). In mPFC and OFC, but not the DS, representation of the transfer event correlated with the strength of PIT behavior. Neurons in all three regions showed CS-selective amplification of Pavlovian approaches toward the reward delivery site. Whereas striatal neurons represented transfer and approach behavior through mostly segregated neuronal subsets, overlapping subsets represented these events in the mPFC and OFC. These findings suggest that parallel phasic activation of mPFC and OFC neuronal subsets participates in the transfer from Pavlovian incentives to instrumental actions. PMID- 19309321 TI - Interference of working memory load with long-term memory formation. AB - Traditionally, it has been assumed that the medial temporal lobe (MTL) is indispensable for long-term memory (LTM) encoding, but only plays a minor role for working memory (WM) maintenance. Recently, however, an increasing number of studies questioned this seemingly clear distinction by showing that the MTL does participate in some WM processes, especially if multiple items are being maintained. This would predict that WM maintenance of multiple items interferes with simultaneous LTM encoding. Here, we tested this idea in a functional magnetic resonance imaging paradigm that required subjects to encode stimuli into LTM during simultaneous WM maintenance of either single or multiple items. Indeed, we found that maintenance of multiple items deteriorates simultaneous LTM encoding as compared with maintenance of single items. WM-related activation of the hippocampus was more pronounced in the condition with high WM load; in contrast, hippocampal activation related to LTM encoding was stronger in the low WM load condition. Successful LTM encoding was associated with a high level of activity in the adjacent parahippocampal cortex (PHC), leading to pronounced parahippocampal subsequent memory effects in the high load condition. This suggests that the PHC is a locus of WM-LTM interaction. Functional connectivity analysis with a seed in the PHC confirmed this result by revealing strong connectivity with the medial frontal cortex, which was only active in the high WM load condition. Taken together, these findings suggest that high WM demands interfere with LTM encoding and thus support the idea that WM and LTM processes interact in the MTL. PMID- 19309322 TI - Comprehensive analysis of cooperative gene mutations between class I and class II in de novo acute myeloid leukemia. AB - Acute myeloid leukemia (AML) has been thought to be the consequence of two broad complementation classes of mutations: class I and class II. However, overlap mutations between them or within the same class and the position of TP53 mutation are not fully analyzed. We comprehensively analyzed the FLT3, cKIT, N-RAS, C/EBPA, AML1, MLL, NPM1, and TP53 mutations in 144 newly diagnosed de novo AML. We found 103 of 165 identified mutations were overlapped with other mutations, and most overlap-mutations consisted of class I and class II mutations. Although overlap-mutations within the same class were found in seven patients, five of them additionally had the other class mutation. These results suggest that most overlap-mutations within the same class might be the consequence of acquiring an additional mutation after the completion both of class I and class II mutations. However, mutated genes overlapped with the same class were limited in N-RAS, TP53, MLL-PTD, and NPM1, suggesting the possibility that these irregular overlap mutations might cooperatively participate in the development of AML. Notably, TP53 mutation was overlapped with both class I and class II mutations, and associated with morphologic multilineage dysplasia and complex karyotype. The genotype consisting of complex karyotype and TP53 mutation was an unfavorable prognostic factor in entire AML patients, indicating this genotype generates a disease entity in de novo AML. These results collectively suggest that TP53 mutation might be a functionally distinguishable class of mutation. PMID- 19309323 TI - Characterization of NVX-207, a novel betulinic acid-derived anti-cancer compound. AB - BACKGROUND: Development of betulinic acid derivatives for clinical use has been hampered by adverse pharmacological and physico-chemical characteristics of this class of compounds. We here present a novel semi-synthetic betulinic acid-derived drug candidate well suited for further clinical development. MATERIALS AND METHODS: In vitro activity and mode of action of NVX-207 were determined using normal as well as cancer cell lines. Gene expression profiling was performed with Affymetrix U133 microarrays. NVX-207 binding partners were identified using a heterobifunctional chemical crosslinker system. Potential binding proteins were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) analysis. Clinical studies were conducted in canine cancer patients suffering from spontaneously arising pre-treated tumours. RESULTS: NVX-207 showed anti-tumour activity (mean IC(50) = 3.5 microM) against various human and canine cell lines. NVX-207-induced apoptosis was associated with activation of the intrinsic apoptotic pathway via cleavage of caspases -9, -3, -7 and of poly (ADP ribose) polymerase (PARP). Global gene expression profiling demonstrated regulation of genes associated with lipid metabolism, most notably an upregulation of genes coding for insulin-induced gene 1 (Insig-1), low-density lipoprotein receptor (LDL-R) and of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA). NVX-207 bound to apolipoprotein A-I, a major regulator of lipid metabolism and cholesterol transport. A phase I/II study in dogs suffering from naturally occurring cancer receiving local treatment of NVX-207 (10 mg mL(-1)) showed excellent clinical responses including a complete remission in so far 5/5 treated animals. CONCLUSIONS: NVX-207 is well tolerated and has significant anti-cancer activity in vitro and in vivo in dogs with treatment-resistant malignancies. PMID- 19309324 TI - Mood irritability - do we need to refine the diagnostic validity of oppositional defiant disorder and paediatric bipolar disorder? PMID- 19309325 TI - Practitioner review: the assessment of bipolar disorder in children and adolescents. AB - BACKGROUND: An increasing number of youth are being diagnosed with, and treated for, bipolar disorder (BD). Controversy exists about whether youth with non episodic irritability and symptoms of attention deficit hyperactivity disorder (ADHD) should be considered to have a developmental presentation of mania. METHOD: A selective review of the literature related to this question, along with recommendations to guide clinical assessment. RESULTS: Data indicate differences between youth with episodic mania and those with non-episodic irritability in longitudinal diagnostic associations, family history, and pathophysiology. In youth with episodic mania, elation and irritability are both common during manic episodes. CONCLUSIONS: In diagnosing mania in youth, clinicians should focus on the presence of episodes that consist of a distinct change in mood accompanied by concurrent changes in cognition and behavior. BD should not be diagnosed in the absence of such episodes. In youth with ADHD, symptoms such as distractibility and agitation should be counted as manic symptoms only if they are markedly increased over the youth's baseline symptoms at the same time that there is a distinct change in mood and the occurrence of other associated symptoms of mania. Although different techniques for diagnosing comorbid illnesses have not been compared systematically, it appears most rational to diagnose co-occurring illnesses such as ADHD only if the symptoms of the co-occurring illness are present when the youth is euthymic. PMID- 19309326 TI - Cognitive behavioral therapy for anxiety in children with autism spectrum disorders: a randomized, controlled trial. AB - BACKGROUND: Children with autism spectrum disorders often present with comorbid anxiety disorders that cause significant functional impairment. This study tested a modular cognitive behavioral therapy (CBT) program for children with this profile. A standard CBT program was augmented with multiple treatment components designed to accommodate or remediate the social and adaptive skill deficits of children with ASD that could pose barriers to anxiety reduction. METHOD: Forty children (7-11 years old) were randomly assigned to 16 sessions of CBT or a 3 month waitlist (36 completed treatment or waitlist). Therapists worked with individual families. The CBT model emphasized behavioral experimentation, parent training, and school consultation. Independent evaluators blind to treatment condition conducted structured diagnostic interviews and parents and children completed anxiety symptom checklists at baseline and posttreatment/postwaitlist. RESULTS: In intent-to-treat analyses, 78.5% of the CBT group met Clinical Global Impressions-Improvement scale criteria for positive treatment response at posttreatment, as compared to only 8.7% of the waitlist group. CBT also outperformed the waitlist on diagnostic outcomes and parent reports of child anxiety, but not children's self-reports. Treatment gains were maintained at 3 month follow-up. CONCLUSIONS: The CBT manual employed in this study is one of the first adaptations of an evidence-based treatment for children with autism spectrum disorders. Remission of anxiety disorders appears to be an achievable goal among high-functioning children with autism. PMID- 19309327 TI - The effects of foster care intervention on socially deprived institutionalized children's attention and positive affect: results from the BEIP study. AB - BACKGROUND: We examined the effects of a foster care intervention on attention and emotion expression in socially deprived children in Romanian institutions. METHODS: Institutionalized children were randomized to enter foster care or to remain under institutional care. Subsequently, the institutionalized and foster care groups, along with a community-based comparison group, were evaluated on emotion tasks at 30 and 42 months of age. Behaviors reflecting positive and negative affect and attention were coded from videotapes. RESULTS: Data indicated that at both age points, children who received the foster care intervention showed higher levels of attention and positive affect compared to children who remained institutionalized. Compared to the community sample, children in the foster care intervention showed higher levels of attention to the emotion eliciting tasks at 42 months of age. CONCLUSIONS: The results of this randomized trial demonstrate the impact of a family-based intervention on the development of attention and positive affect. PMID- 19309328 TI - Maternal stress and depression and the lateralisation of infant cradling. AB - BACKGROUND: Studies show that 65-85% of mothers cradle their infants to the left side of their body, but that this bias changes with maternal mood and stress. The present study examines the hypothesis that maternal stress and depression status will influence the cradling bias differentially. METHOD: As part of a larger study on mother-infant interaction, mothers (N = 79) were asked to pick up and briefly hold their children in their arms (44 boys, 35 girls; mean age 7.2 months, range 3 to 14 months). RESULTS: Results indicated that 86% of mothers who were neither stressed nor depressed cradled to the left and 14% to the right. Comparing the cradling side of stressed mothers with those who were neither stressed nor depressed, more in the former group showed right-sided cradling. In contrast, mothers who were just depressed preferred to cradle to the left. CONCLUSION: The lack of a left-sided cradling bias might be due to stress rather than depression experienced by mothers. Furthermore, this study provides evidence that the state of maternal mental health might be indicated by the side on which they cradle their child preferentially. PMID- 19309329 TI - Reduced change blindness suggests enhanced attention to detail in individuals with autism. AB - BACKGROUND: The phenomenon of change blindness illustrates that a limited number of items within the visual scene are attended to at any one time. It has been suggested that individuals with autism focus attention on less contextually relevant aspects of the visual scene, show superior perceptual discrimination and notice details which are often ignored by typical observers. METHODS: In this study we investigated change blindness in autism by asking participants to detect continuity errors deliberately introduced into a short film. Whether the continuity errors involved central/marginal or social/non-social aspects of the visual scene was varied. Thirty adolescent participants, 15 with autistic spectrum disorder (ASD) and 15 typically developing (TD) controls participated. RESULTS: The participants with ASD detected significantly more errors than the TD participants. Both groups identified more errors involving central rather than marginal aspects of the scene, although this effect was larger in the TD participants. There was no difference in the number of social or non-social errors detected by either group of participants. CONCLUSION: In line with previous data suggesting an abnormally broad attentional spotlight and enhanced perceptual function in individuals with ASD, the results of this study suggest enhanced awareness of the visual scene in ASD. The results of this study could reflect superior top-down control of visual search in autism, enhanced perceptual function, or inefficient filtering of visual information in ASD. PMID- 19309330 TI - Comparison of symptomatic versus functional changes in children and adolescents with ADHD during randomized, double-blind treatment with psychostimulants, atomoxetine, or placebo. AB - BACKGROUND: This meta-analysis was designed to determine the relationship between reduction of attention-deficit/hyperactivity disorder (ADHD) symptoms and improvement in functioning by examining short-term changes in functional and symptomatic scores in children and adolescents with ADHD. METHODS: Search of atomoxetine's clinical trial database identified four studies involving a symptomatic measure, the ADHD Rating Scale-IV-Parent Version:Investigator administered and -scored (ADHDRS-IV-Parent:Inv), and a functional measure, the Life Participation Scale for ADHD (LPS). RESULTS: Correlation analysis revealed a moderate-to-strong association between changes in the LPS total versus ADHDRS-IV Parent:Inv total (r: -.68). The LPS Self-control subscale showed higher correlations than the Happy/Social subscale with the symptomatic measures. Regression analysis also showed high sensitivity for functional measures to changes in symptom severity. Stratified analysis of mean changes in ADHDRS-IV Parent:Inv scores corresponding to standardized changes in LPS functional scores indicated that a threshold reduction of 16-18 points on the ADHDRS-IV-Parent:Inv total score was needed for functional improvements to become evident. CONCLUSIONS: Subjects' symptomatic improvements appear to be reflected in improvements in their social and behavioral function as measured by the LPS. These initial findings warrant verification by replication with other outcome measures. PMID- 19309331 TI - Familial risk analysis of the association between attention-deficit/hyperactivity disorder and psychoactive substance use disorder in female adolescents: a controlled study. AB - BACKGROUND: A robust and bi-directional comorbidity between attention deficit/hyperactivity disorder (ADHD) and psychoactive substance use disorder (PSUD, alcohol or drug abuse, or dependence) has been consistently reported in the literature. However, this literature has been based almost exclusively on male only samples and, therefore, the findings may not generalize to females. METHODS: First-degree relatives from a large sample of pediatrically and psychiatrically referred girls with (123 probands, 403 relatives) and without ADHD (112 probands, 359 relatives) were comprehensively assessed by blind raters with structured diagnostic interviews. Familial risk analysis examined the risks in first-degree relatives for ADHD and PSUD (alcohol or drug abuse or dependence) after stratifying probands by the presence and absence of these disorders. RESULTS: ADHD in the proband significantly increased the risk for ADHD in relatives independently of the comorbidity with PSUD. PSUD in the proband was associated with a significantly increased risk for PSUD in relatives regardless of ADHD status. There was no evidence of co-segregation or non-random mating in the families of probands with ADHD and PSUD. CONCLUSIONS: Patterns of familial risk analysis suggest that the association between ADHD and PSUD in adolescent females is most consistent with the hypothesis that these disorders are independently transmitted, although the hypothesis of variable expressivity could not be ruled out. These findings are consistent with previously reported patterns of familial associations between ADHD and PSUD found in adolescent males. Longer follow-up periods are needed to more fully clarify the relationship between ADHD and PSUD, as well as provide adequate power for separate analyses of alcohol and drug use. PMID- 19309336 TI - Comment on Nilsson et al., tumefactive demyelinating disease treated with decompressive craniectomy. PMID- 19309337 TI - Tumefactive demyelinating disease treated with decompressive craniectomy. AB - BACKGROUND: Tumefactive demyelinating disease (TDD) is a rare primary demyelinating disease with diagnostic and therapeutic challenges. METHODS AND RESULTS: We report a 50-year old woman with TDD successfully treated with decompressive craniectomy and corticosteroids. The patient presented with seizures, subacute progressive hemispheric syndrome, and a tumourlike abnormality on MRI. Demyelinating disease was initially considered unlikely. Due to a rapidly evolving herniation syndrome hemicraniectomy was performed. Outcome was favourable with only very mild neurological deficits 6 weeks later. CONCLUSION: TDD should be considered as a differential diagnosis in tumour-like presentations, and appears to have distinctive neuroimaging features. In the advent of treatement failure from high dose corticosteroids and plasmapheresis and development of severe mass effect, decompressive hemicraniectomy is an important treatment option. PMID- 19309338 TI - Photodynamic therapy for the treatment of cutaneous neoplasia, inflammatory disorders, and photoaging. AB - BACKGROUND: Photodynamic therapy (PDT) has demonstrated high efficacy, minimal side effects, and improved cosmetic outcome when used for the treatment of actinic keratoses (AK), basal cell carcinoma (BCC), squamous cell carcinoma, and photoaging. METHODS: To review the literature on the use of PDT in dermatologic surgery using MEDLINE. RESULTS: Published clinical studies using PDT in the treatment of AKs yield overall efficacy rates ranging from 50% to 71% with one treatment to as high as 88% to 90% with two or more treatments. For superficial BCC, initial clearance rates were 76% to 97%, and for Bowen's disease, initial clearance rates ranged from 72% to 94% overall. The use of PDT for photorejuvenation is a relatively new application of this technology, which has shown promise in improving the appearance of fine lines, pigmentary variation, and telangiectasias. CONCLUSIONS: The advantages of photodynamic therapy include the capacity for noninvasive targeted therapy through topical application of aminolevulinic acid and methyl aminolevulinic acid, with outstanding cosmetic results. Although the theory behind the use of chemical photosensitizers and ultraviolet light to treat a wide variety of skin disorders is straightforward, the practical application of this technology is evolving. Additional research into the precise mechanisms of action for specific photosensitizers and optimal light sources will be highly beneficial to the advancement of this technology. PMID- 19309339 TI - Epithelioid sarcoma: a diagnostic and surgical challenge. PMID- 19309340 TI - The lateral ala's volume and position are critical determinants of aesthetically successful nasal reconstruction: a photographic case series. PMID- 19309341 TI - Keratoacanthomatous hyperplasia in response to a tattoo. PMID- 19309342 TI - A modified technique for processing Mohs wedge excisions. PMID- 19309343 TI - Intractable wounds caused by pyoderma gangrenosum in a patient with critical limb ischemia treated with cyclosporine and adjuvant sympathectomy. PMID- 19309344 TI - A novel specialized suture and inserting device for the resuspension of ptotic facial tissues: early results. AB - BACKGROUND: In the past decade, the popularity of minimally invasive procedures for facial rejuvenation has increased. OBJECTIVE: To describe a new specialized suture, and its associated technique, used to elevate sagging tissues of the face and neck. METHODS: A detailed description of the technique and the results obtained in 20 patients in whom we have used this novel approach. Attention was given to appropriate patient selection. The primary focus was on the correction of the jowl, jawline, and neck subunits. It involves the percutaneous introduction of a novel 3-0 polypropylene suture that has 10 absorbable hollow cones along its axis that are equally interspersed with knots. Once the absorbable cones are resorbed into the surrounding tissues, the non-absorbable suture component can be removed without compromising the aesthetic outcome. RESULTS: All patients demonstrated improvement in these areas, with minimal complications. One patient required resuspension using the open technique. (Excessive ptotic tissue was later excised for an optimal cosmetic result.) CONCLUSION: The suture and technique described in this article provide a major contribution to the correction of ptosis of facial tissues. When done in conjunction with other procedures, such as neck and jowl microliposuction, this technique has proven to be a useful addition to facial rejuvenation. PMID- 19309345 TI - Lentigo maligna--outcomes of treatment with Q-switched Nd:YAG and alexandrite lasers. AB - BACKGROUND: Surgical excision is considered the criterion standard treatment for lentigo maligna (LM) but may sometimes be unsuitable for elderly people or in the treatment of large lesions. OBJECTIVE: To evaluate the role of Q-switched neodymium-doped yttrium aluminium garnet (QS:Nd:YAG) and alexandrite lasers in the treatment of LM. METHODS: QS:NdYAG and alexandrite lasers were used to treat histologically proven LM in 22 patients who were considered unsuitable for wide surgical excision or declined it. Patients were assessed at 6-month intervals for a maximum period of 5 years. RESULTS: Complete clinical response was achieved in 12 patients after one to four treatments and a follow-up of 2 to 5 years after last treatment. Excellent cosmesis was achieved in all patients. LM melanoma developed in two lesions, which were subsequently excised. Recurrence occurred in four patients. CONCLUSION: Although QS:Nd:YAG and alexandrite laser treatment of LM produced long-term clearance in 12 of the 22 patients, the ease and speed of this treatment and excellent cosmetic outcome achieved make this a suitable alternative to surgical excision, especially in elderly people, in treatment of large lesions, or in patients who refuse surgical treatment. PMID- 19309346 TI - Treatment of inflammatory facial acne with 1,450-nm diode laser in type IV to V Asian skin using an optimal combination of laser parameters. AB - BACKGROUND: The 1,450-nm diode laser is effective for the treatment of inflammatory acne, but there is a significant risk of postinflammatory hyperpigmentation (PIH) in Asian skin. OBJECTIVE: To determine whether lower fluence and shorter cooling duration of the 1,450-nm diode laser improve acne effectively with minimal PIH in darker skin. MATERIALS AND METHODS: Twenty-six subjects (skin phototypes IV-V) with inflammatory facial acne received four treatments using the 1,450-nm diode laser with 6-mm spot size 3 to 4 weeks apart. We used three passes with a fluence of 8 J/cm(2) with dynamic cooling of 25 ms to minimize PIH. Serial blinded assessment of acne lesion counts and sebum measurement were evaluated before and up to 6 months after treatment. RESULTS: Four weeks and 6 months after the last treatment, reduction of mean acne lesions was 29% (p<.01) and 40% (p<.03), respectively, from baseline in the group with moderate acne. Significant improvements of sebum production were noted. Four episodes of temporary PIH (3.8%) were observed out of all treatment sessions. CONCLUSION: Use of multiple passes of a 1,450-nm diode laser with lower fluence and shorter dynamic cooling device retains its therapeutic efficacy with substantial reduction of PIH for moderate acne. PMID- 19309347 TI - Topical methyl aminolevulinate photodynamic therapy using red light-emitting diode light for multiple actinic keratoses: a randomized study. AB - BACKGROUND: Photodynamic therapy (PDT) is an effective treatment for actinic keratoses (AKs). Light-emitting diodes (LEDs) offer practical advantages when treating multiple lesions. OBJECTIVE: To evaluate the efficacy and tolerability of PDT using a LED and topical methyl aminolevulinate (MAL) for treatment of multiple AKs. METHODS AND MATERIALS: One hundred thirty-one patients with four to 10 non-pigmented, previously untreated thin or moderately thick AKs on the face or scalp were enrolled in this multicenter, double-blind, randomized, placebo controlled study. MAL or matching placebo cream was applied to the debrided lesion surface for 3 hours before illumination with noncoherent red light (630 nm, light dose 37 J/cm(2)). Treatment was repeated 1 week later. RESULTS: Efficacy was evaluated in 57 patients with 418 lesions treated with MAL PDT and 58 with 414 lesions treated with placebo PDT. Sixteen patients were excluded as protocol violators (not randomized). MAL PDT was superior (p< .001) to placebo PDT in lesion complete response rates (83.3%, 95% confidence interval (CI)=79.3 86.7%, vs 28.7%, 95% CI=24.4-33.4%) and patient complete response rates (all lesions showing complete response; 68.4%, 95% CI=54.8-80.1% vs 6.9%, 95% CI=1.9 16.7%). CONCLUSIONS: Topical MAL PDT using a LED is an effective treatment for multiple AKs. PMID- 19309348 TI - Translational mini-review series on immunology of vascular disease: mechanisms of vascular inflammation and remodelling in systemic vasculitis. AB - Vessel walls are the primary inflammatory sites in systemic vasculitides. In most cases the initiating event is unknown, and a self-sustaining circuit attracts and activates inflammatory leucocytes in the wall of vessels of various size and anatomical characteristics. Recent studies have revealed homeostatic roles of vascular inflammation and have identified the action of humoral innate immunity, in particular injury-associated signals and acute phase proteins, on the activation of circulating leucocytes, platelets and endothelial cells. These advances have provided clues to the molecular mechanisms underlying the vicious circle that maintains and amplifies vessel and tissue injury. PMID- 19309349 TI - Translational mini-review series on immunology of vascular disease: inflammation, infections and Toll-like receptors in cardiovascular disease. AB - Cardiovascular disease, in which atherosclerosis is the major underlying cause, is currently the largest cause of death in the world. Atherosclerosis is an inflammatory disease characterized by the formation of arterial lesions over a period of several decades at sites of endothelial cell dysfunction. These lesions are composed of endothelial cells, vascular smooth muscle cells, monocytes/macrophages and T lymphocytes (CD4(+)). As the lesions progress some can become unstable and prone to disruption, resulting in thrombus formation and possibly a myocardial infarction or stroke depending upon the location. Although the exact triggers for plaque disruption remain unknown, much recent evidence has shown a link between the incidence of myocardial infarction and stroke and a recent respiratory tract infection. Interestingly, many reports have also shown a link between a family of pattern recognition receptors, the Toll-like receptors, and the progression of atherosclerosis, suggesting that infections may play a role in both the progression of atherosclerosis and in inducing the more severe complications associated with the disease. PMID- 19309350 TI - Translational mini-review series on immunology of vascular disease: accelerated atherosclerosis in vasculitis. AB - Premature atherosclerosis has been observed during the course of different systemic inflammatory diseases such as rheumatoid arthritis and sytemic lupus erythematosus. Remarkably, relatively few studies have been published on the occurrence of accelerated atherosclerosis in patients with vasculitis. In giant cell arteritis (GCA), mortality because of ischaemic heart disease is not increased. In addition, intima media thickness (IMT) is lower in patients with GCA than in age-matched controls. In contrast, IMT is increased significantly in Takayasu arteritis, another form of large vessel vasculitis occurring in younger patients. In Takayasu arteritis and in Kawasaki disease, a form of medium-sized vessel vasculitis, accelerated atherosclerosis has been well documented. In small vessel vasculitis because of anti-neutrophil cytoplasmic autoantibodies associated vasculitis, cardiovascular diseases are a major cause of mortality. IMT measurements reveal conflicting results. During active disease these patients experience acceleration of the atherosclerotic process. However, when inflammation is controlled, these patients have atherosclerotic development as in healthy subjects. Several risk factors, such as diabetes and hypertension, are present more often in patients with vasculitis compared with healthy controls. In addition, steroids may be pro-atherogenic. Most importantly, many patients have impaired renal function, persistent proteinuria and increased levels of C reactive protein, well-known risk factors for acceleration of atherosclerosis. Enhanced oxidation processes, persistently activated T cells and reduced numbers of regulatory T cells are among the many pathophysiological factors that play a role during acceleration of atherogenesis. Finally, autoantibodies that may be relevant for acceleration of atherosclerosis are found frequently in elevated titres in patients with vasculitis. Because patients have an increased risk for cardiovascular events, vasculitis should be treated with as much care as possible. In addition, treatment should be considered with angiotensin-converting enzyme inhibitors and/or angiotensin receptor-1 blockers, statins and acetylsalicyl acid. Finally, classical risk factors for cardiovascular disease should be monitored and treated as much as possible. PMID- 19309351 TI - Opposite effect of fluticasone and salmeterol on fibronectin and tenascin-C expression in primary human lung fibroblasts. AB - BACKGROUND: Airway remodelling is a key feature of asthma and chronic obstructive pulmonary disease (COPD). The remodelling process involves the deposition of extracellular matrix (ECM) proteins within the airways. Current therapies for asthma and COPD consist of inhaled corticosteroids and long-acting beta(2) agonists (LABA). However, their effect on airway remodelling is not well understood so far. OBJECTIVE: In this study we investigated the effect of fluticasone and salmeterol, either alone or in combination, on fibronectin and tenascin-C protein, isoform, and mRNA levels in primary human lung fibroblasts. METHODS: In our model, fibroblasts cultured in serum-free medium represented a non-inflammatory condition and stimulation with 5% fetal calf serum and/or TGF beta(1) mimicked a pro-fibrotic environment with activation of tissue repair. Using these two different conditions, the effects of fluticasone and salmeterol on fibronectin and tenascin-C protein and mRNA levels were analysed by immunoblotting and semi-quantitative RT-PCR. RESULTS: In both conditions, fluticasone increased fibronectin transcript and protein levels, whereas it decreased those of tenascin-C. Salmeterol neither affected fibronectin and tenascin-C synthesis significantly nor did it influence the effect of fluticasone when applied in combination. Furthermore, we found that treatment with fluticasone had an opposite effect on extra domain A and B containing fibronectin isoforms generated by alternative splicing compared with total fibronectin transcript levels, whereas tenascin-C isoforms were not differently modulated by fluticasone. CONCLUSIONS: Our results indicate that standard therapies for inflammatory lung disorders influence ECM protein composition and relative expression levels. In contrast to corticosteroids, LABA did not significantly alter the expression of tenascin-C and fibronectin in cultures of primary human lung fibroblasts. PMID- 19309352 TI - Suppressor of cytokine signalling 1 (SOCS1) is a physiological regulator of the asthma response. AB - BACKGROUND: The molecular determinants of the severity and persistence of allergic asthma remain poorly understood. Suppressor of cytokine signalling 1 (SOCS1) is a negative regulator of IL-4-dependent pathways in vitro and might therefore control T-helper type 2 (Th2) immunity associated traits, such as IgE levels, mucin production, IL-5 and IL-13 induction, and eosinophilic mucosal inflammation, which are implicated in allergic asthma. OBJECTIVE: To investigate the role of SOCS1 in regulating Th2-associated disease traits in a murine sub chronic aeroallergen-driven asthma model. METHODS: Following sensitization and challenge with ovalbumin (OVA), bronchoalveolar lavage and serum were collected from mice lacking the Socs1 gene on an IFN-gamma null background (Socs1(-/ )Ifngamma(-/-)). The composition of infiltrating cells in the lung, serum IgE and IgG1 levels and cytokine levels were analysed. RESULTS: Serum IgE levels and infiltrating eosinophils were considerably increased in the lungs of OVA-treated Socs1(-/-)Ifngamma(-/-) mice compared with Ifngamma(-/-) and C57BL/6 controls. Expression of the Th2 cytokines, IL-4, IL-5 and IL-13 was increased in CD4+ cells and lung tissue from OVA-treated Socs1(-/-)Ifngamma(-/-) mice. IgE, IL-5 levels and infiltrating eosinophils were also elevated in saline-treated Socs1(-/ )Ifngamma(-/-) mice, suggesting that in the absence of SOCS1, mice are already biased towards a Th2 response. It is at present unclear whether the elevated cytokine levels are sufficient to result in the exacerbated Th2 response to OVA challenge or whether enhanced intra-cellular signalling also contributes. Surprisingly, of the various IL-4/IL-13 responsive genes tested, only Arginase I appeared to be modestly up-regulated in the lungs of OVA-treated Socs1(-/ )Ifngamma(-/-) mice, suggesting that regulation by SOCS1 occurs primarily in haematopoietic cells and not in the airway epithelium. CONCLUSIONS: Together these results indicate that SOCS1 is an important regulator of the Th2 response. PMID- 19309353 TI - G protein-coupled receptor hetero-dimerization: contribution to pharmacology and function. AB - The concept that G protein-coupled receptors (GPCRs) can form hetero-dimers or hetero-oligomers continues to gain experimental support. However, with the exception of the GABA(B) receptor and the sweet and umami taste receptors few reported examples meet all of the criteria suggested in a recent International Union of Basic and Clinical Pharmacology sponsored review (Pin et al., 2007) that should be required to define distinct and physiologically relevant receptor species. Despite this, there are many examples in which pairs of co-expressed GPCRs reciprocally modulate their function, trafficking and/or ligand pharmacology. Such data are at least consistent with physical interactions between the receptor pairs. In recent times, it has been suggested that specific GPCR hetero-dimer or hetero-oligomer pairs may represent key molecular targets of certain clinically effective, small molecule drugs and there is growing interest in efforts to identify ligands that may modulate hetero-dimer function selectively. The current review summarizes key recent developments in these topics. PMID- 19309354 TI - The role of histamine H4 receptor in immune and inflammatory disorders. AB - Since its discovery at the beginning of the 20th century, histamine has been established to play a pathophysiological regulatory role in cellular events through binding to four types of G-protein-coupled histamine receptors that are differentially expressed in various cell types. The discovery, at the turn of the millennium, that the histamine H4 receptor is largely expressed in haemopoietic cells as well as its chemotactic properties designate its regulatory role in the immune system. H4 receptors modulate eosinophil migration and selective recruitment of mast cells leading to amplification of histamine-mediated immune responses and eventually to chronic inflammation. H4 receptor involvement in dendritic cell activation and T cell differentiation documents its immunomodulatory function. The characterization of the H4 as the immune system histamine receptor directed growing attention towards its therapeutic exploitation in inflammatory disorders, such as allergy, asthma, chronic pruritus and autoimmune diseases. The efficacy of a number of H4 receptor ligands has been evaluated in in vivo and in vitro animal models of disease and in human biological samples. However, before reaching decisive conclusions on H4 receptor pathophysiological functions and therapeutic exploitation, identification of genetic polymorphisms and interspecies differences in its relative actions and pharmacological profile need to be addressed and taken into consideration. Despite certain variations in the reported findings, the available data strongly point to the H4 receptor as a novel target for the pharmacological modulation of histamine-transferred immune signals and offer an optimistic perspective for the therapeutic exploitation of this promising new drug target in inflammatory disorders. PMID- 19309355 TI - 5-hydroxytryptamine induced relaxation in the pig urinary bladder neck. AB - BACKGROUND AND PURPOSE: 5-Hydroxytryptamine (5-HT) is one of the inhibitory mediators in the urinary bladder outlet region. Here we investigated mechanisms involved in 5-HT-induced relaxations of the pig bladder neck. EXPERIMENTAL APPROACH: Urothelium-denuded strips of pig bladder were mounted in organ baths for isometric force recordings of responses to 5-HT and electrical field stimulation (EFS). KEY RESULTS: After phenylephrine-induced contraction, 5-HT and 5-HT receptor agonists concentration-dependently relaxed the preparations, with the potency order: 5-carboxamidotryptamine (5-CT) > 5-HT = RS67333 > (+/-)-8 hydroxy-2-dipropylaminotetralinhydrobromide > m-chlorophenylbiguanide > alpha methyl-5-HT > ergotamine. 5-HT and 5-CT relaxations were reduced by the 5-HT(7) receptor antagonist (2R)-1-[(3-hydroxyphenyl)sulphonyl]-2-[2-(4-methyl-1 piperidinyl)ethyl]pyrrolidine hydrochloride and potentiated by (S)-N-tert-butyl-3 (4-(2-methoxyphenyl)-piperazin-1-yl)-2-phenylpropanamide dihydrochloride (WAY 100135) and cyanopindolol, 5-HT(1A) and 5-HT(1A/1B) receptor antagonists respectively. Inhibitors of 5-HT(1B/1D), 5-HT(2), 5-HT(2B/2C), 5-HT(3), 5-HT(4), 5-HT(5A) and 5-HT(6) receptors failed to modify 5-HT responses. Blockade of monoamine oxidase A/B, noradrenergic neurotransmission, alpha-adrenoceptors, muscarinic and purinergic receptors, nitric oxide synthase, guanylate cyclase and prostanoid synthesis did not alter relaxations to 5-HT. Inhibitors of Ca(2+) activated K(+) and ATP-dependent K(+) channels failed to modify 5-HT responses but blockade of neuronal voltage-gated Na(+)-, Ca(2+)- and voltage-gated K(+) (K(v))-channels potentiated these relaxations. Adenylyl cyclase activation and cAMP-dependent protein kinase (PKA) inhibition potentiated and reduced, respectively, 5-HT-induced responses. Under non-adrenergic, non-cholinergic, non nitrergic conditions, EFS induced neurogenic, frequency-dependent, relaxations which were resistant to WAY 100135 and cyanopindolol. CONCLUSIONS AND IMPLICATIONS: 5-HT relaxed the pig urinary bladder neck through muscle 5-HT(7) receptors linked to the cAMP-PKA pathway. Prejunctional 5-HT(1A) receptors and K(v) channels modulated 5-HT-induced relaxations whereas postjunctional K(+) channels were not involved in such responses. 5-HT(7) receptor antagonists could be useful in the therapy of urinary incontinence produced by intrinsic sphincter deficiency. PMID- 19309356 TI - Activation of iGluR5 kainate receptors inhibits neurogenic dural vasodilatation in an animal model of trigeminovascular activation. AB - BACKGROUND AND PURPOSE: Migraine is a disabling neurological disorder involving activation, or the perception of activation, of trigeminovascular afferents containing calcitonin gene-related peptide (CGRP). Released CGRP from peripheral trigeminal afferents causes dilatation of dural blood vessel, and this is used to measure trigeminal nerve activation. Kainate receptors with the GluR5 subunit (iGluR5, ionotropic glutamate receptor) are present in the trigeminal ganglion and may be involved in nociception. We investigated the possible involvement of prejunctional iGluR5 kainate receptors on CGRP release from trigeminal afferents. EXPERIMENTAL APPROACH: We used neurogenic dural vasodilatation, which involves reproducible vasodilatation in response to CGRP release after electrical stimulation of the dura mater surrounding the middle meningeal artery. The effects of the specific iGluR5 receptor antagonist UBP 302 and agonist (S)-(-)-5 iodowillardiine were investigated on neurogenic and CGRP-induced dural vasodilatation in rats, by using intravital microscopy. KEY RESULTS: Administration of 10 and 20 mg.kg(-1) of iodowillardiine inhibited electrically induced dural vessel dilatation, an effect blocked by pretreatment with 50 mg.kg( 1) UBP 302. Administration of the iGluR5 receptor antagonist UBP 302 alone had no significant effect. CGRP (1 mg.kg(-1))-induced dural vasodilatation was not inhibited by the iGluR5 receptor agonist iodowillardiine. CONCLUSIONS AND IMPLICATIONS: This study demonstrates that activation of the iGluR5 kainate receptors with the selective agonist iodowillardiine is able to inhibit neurogenic dural vasodilatation probably by inhibition of prejunctional release of CGRP from trigeminal afferents. Taken together with recent clinical studies the data reinforce CGRP mechanisms in primary headaches and demonstrate a novel role for kainate receptor modulation of trigeminovascular activation. PMID- 19309357 TI - Involvement of PKC alpha and G-protein-coupled receptor kinase 2 in agonist selective desensitization of mu-opioid receptors in mature brain neurons. AB - BACKGROUND AND PURPOSE: The ability of an agonist to induce desensitization of the mu-opioid receptor (MOR) depends upon the agonist used. Furthermore, previous data suggest that the intracellular mechanisms underlying desensitization may be agonist-specific. We investigated the mechanisms underlying MOR desensitization, in adult mammalian neurons, caused by morphine (a partial agonist in this system) and DAMGO (a high-efficacy agonist). EXPERIMENTAL APPROACH: MOR function was measured in locus coeruleus neurons, by using whole-cell patch-clamp electrophysiology, in rat and mouse brain slices (both wild-type and protein kinase C (PKC)alpha knockout mice). Specific isoforms of PKC were inhibited by using inhibitors of the receptors for activated C-kinase (RACK), and in vivo viral-mediated gene-transfer was used to transfect neurons with dominant negative mutants (DNMs) of specific G-protein-coupled receptor kinases (GRKs). KEY RESULTS: Morphine-induced desensitization was attenuated by using RACK inhibitors that inhibit PKCalpha, but not by other isoform-specific inhibitors. Further, the PKC component of morphine-induced desensitization was absent in locus coeruleus neurons from PKCalpha knockout mice. The PKC-enhanced morphine-induced desensitization was not affected by over-expression of a GRK2 dominant negative mutant (GRK2 DNM). In contrast, DAMGO-induced MOR desensitization was independent of PKC activity but was reduced by over-expression of the GRK2 DNM but not by that of a GRK6 DNM. CONCLUSIONS AND IMPLICATIONS: In mature mammalian neurons, different MOR agonists can induce MOR desensitization by different mechanisms, morphine by a PKCalpha-mediated, heterologous mechanism and DAMGO by a GRK mediated, homologous mechanism. These data represent functional selectivity at the level of receptor desensitization. PMID- 19309358 TI - Baicalin attenuates air embolism-induced acute lung injury in rat isolated lungs. AB - BACKGROUND AND PURPOSE: Baicalin has been reported to have anti-inflammatory effects and protect against various tissue injuries. However, the effect of baicalin on air embolism-induced acute lung injury has not been tested yet. EXPERIMENTAL APPROACH: Acute lung injury was induced by infusion of air at a rate of 0.25 mL.min(-1) for 1 min into the pulmonary artery of rat isolated lungs. At the end of the experiment, samples were collected for assessment of lung injury, biochemical analysis and histology. Different doses of baicalin (1, 2 and 4 mg.kg(-1)) were given into the perfusate before air infusion. KEY RESULTS: Air embolism elicited a significant increase in microvascular permeability (K(f)), lung weight gain, wet/dry weight ratio, pulmonary artery pressure and protein concentration in the bronchoalveolar lavage fluid. Levels of the cytokines, tumour necrosis factor alpha and cytokine-induced neutrophil chemoattractant-1 in perfusate, and malondialdehyde levels and myeloperoxidase activities in lung tissue were also significantly increased. In addition, histological examination showed increased neutrophil infiltration in lung tissues. Furthermore, nuclear factor-kappaB activity and degradation of IkappaB-alpha were significantly increased in lungs. Pretreatment of the lungs with baicalin (4 mg.kg(-1)) showed a statistically significant difference in all of the assessed parameters, except for alteration in the pulmonary artery pressure. CONCLUSIONS AND IMPLICATIONS: Our study suggests that baicalin attenuated air embolism-induced acute lung injury and may be considered a useful adjunct drug therapy in this clinical condition. PMID- 19309359 TI - Inhibition of transmitter release from rat sympathetic neurons via presynaptic M(1) muscarinic acetylcholine receptors. AB - BACKGROUND AND PURPOSE: M(2), M(3) and/or M(4) muscarinic acetylcholine receptors have been reported to mediate presynaptic inhibition in sympathetic neurons. M(1) receptors mediate an inhibition of K(v)7, Ca(V)1 and Ca(V)2.2 channels. These effects cause increases and decreases in transmitter release, respectively, but presynaptic M(1) receptors are generally considered facilitatory. Here, we searched for inhibitory presynaptic M(1) receptors. EXPERIMENTAL APPROACH: In primary cultures of rat superior cervical ganglion neurons, Ca(2+) currents were recorded via the perforated patch-clamp technique, and the release of [(3)H] noradrenaline was determined. KEY RESULTS: The muscarinic agonist oxotremorine M (OxoM) transiently enhanced (3)H outflow and reduced electrically evoked release, once the stimulant effect had faded. The stimulant effect was enhanced by pertussis toxin (PTX) and was abolished by blocking M(1) receptors, by opening K(v)7 channels and by preventing action potential propagation. The inhibitory effect was not altered by preventing action potentials or by opening K(v)7 channels, but was reduced by PTX and omega-conotoxin GVIA. The inhibition remaining after PTX treatment was abolished by blockage of M(1) receptors or inhibition of phospholipase C. When [(3)H]-noradrenaline release was triggered independently of voltage-activated Ca(2+) channels (VACCs), OxoM failed to cause any inhibition. The inhibition of Ca(2+) currents by OxoM was also reduced by omega-conotoxin and PTX and was abolished by M(1) antagonism in PTX-treated neurons. CONCLUSIONS AND IMPLICATIONS: These results demonstrate that M(1), in addition to M(2), M(3) and M(4), receptors mediate presynaptic inhibition in sympathetic neurons using phospholipase C to close VACCs. PMID- 19309360 TI - Mechanism of action of species-selective P2X(7) receptor antagonists. AB - BACKGROUND AND PURPOSE: AZ11645373 and N-{2-methyl-5-[(1R, 5S)-9-oxa-3,7 diazabicyclo[3.3.1]non-3-ylcarbonyl]phenyl}-2-tricyclo[3.3.1.13,7]dec-1 ylacetamide hydrochloride (compound-22) are recently described P2X(7) receptor antagonists. In this study we have further characterized these compounds to determine their mechanism of action and interaction with other species orthologues. EXPERIMENTAL APPROACH: Antagonist effects at recombinant and chimeric P2X(7) receptors were assessed by ethidium accumulation and radioligand binding studies. KEY RESULTS: AZ11645373 and compound-22 were confirmed as selective non-competitive antagonists of human or rat P2X(7) receptors respectively. Both compounds were weak antagonists of the mouse and guinea-pig P2X(7) receptors and, for each compound, their potency estimates at human and dog P2X(7) receptors were similar. The potency of compound-22 was moderately temperature-dependent while that of AZ11645373 was not. The antagonist effects of both compounds were slowly reversible and were not prevented by decavanadate, suggesting that they were allosteric antagonists. Indeed, the compounds competed for binding sites labelled by an allosteric radio-labelled P2X(7) receptor antagonist. The species selectivity of AZ11645373, but not compound-22, was influenced by the nature of the amino acid at position 95 of the P2X(7) receptor. N(2)-(3,4-difluorophenyl)-N(1)-[2-methyl-5-(1 piperazinylmethyl)phenyl]glycinamide dihydrochloride, a positive allosteric modulator of the rat receptor, reduced the potency of compound-22 at the rat receptor but had little effect on the actions of AZ11645373. CONCLUSIONS: AZ11645373 and compound-22 are allosteric antagonists of human and rat P2X(7) receptors respectively. The differential interaction of the two compounds with the receptor suggests there may be more than one allosteric regulatory site on the P2X(7) receptor at which antagonists can bind and affect receptor function. PMID- 19309361 TI - Different response patterns of several ligands at the sphingosine-1-phosphate receptor subtype 3 (S1P(3)). AB - BACKGROUND AND PURPOSE: Recently, some ligands targeting the sphingosine-1 phosphate receptor subtype 3 (S1P(3)) have become available. The characterization of these compounds was mainly based on one functional read-out system, although S1P(3) receptors are known to activate different signal transduction pathways. Therefore, this study pharmacologically characterizes these compounds using different assays. EXPERIMENTAL APPROACH: Using CHO-FlpIn cells expressing the human S1P(3) receptor the potencies and maximal effects of S1P, FTY720-P, VPC23019, VPC23153 and VPC24191 were determined in three different assays [inhibition of cAMP accumulation, elevation of intracellular calcium concentrations ([Ca(2+)](i)) and S1P(3) receptor internalization]. KEY RESULTS: All compounds tested inhibited forskolin-induced cAMP accumulation, increased [Ca(2+)](i) and induced S1P(3) receptor internalization but with different potencies and maximal effects. S1P was the most potent compound in all assays followed by FTY720-P. The VPC compounds were generally less potent than S1P and FTY720-P. Regarding the maximal effects, all compounds except VPC23153, behaved as full agonists in the cAMP accumulation assay. In the calcium assay, FTY720-P, VPC23019 and VPC24191 displayed partial and VPC23153 weak partial agonist activity, relative to S1P. Interestingly, treatment with the G(i) inactivator Pertussis toxin, did not affect S1P-induced [Ca(2+)](i) elevations but inhibited those in response to the other compounds, by about 50%. CONCLUSIONS AND IMPLICATIONS: This study demonstrated differential response patterns at the S1P(3) receptor for a range of ligands. These differences could indicate the presence of functional selectivity at this receptor as FTY720-P and the VPC compounds seemed to signal predominantly via G(i)- whereas S1P activated G(i) and G(q)-coupled pathways. PMID- 19309363 TI - Epstein-Barr virus-encoded latent membrane protein 1 activates beta-catenin signaling in B lymphocytes. AB - The Epstein-Barr virus-encoded latent membrane protein 1 is considered the Epstein-Barr virus oncogene based on its importance in Epstein-Barr virus-induced B-lymphocyte transformation. Beta-catenin is a potential oncogene, and its accumulation has been implicated in a variety of human cancers. Here, we found that beta-catenin protein was highly expressed in Epstein-Barr virus-immortalized B-cell lines compared with peripheral blood mononuclear cells from healthy donors. Beta-catenin expression in Epstein-Barr virus-immortalized B-cell line decreased following treatment with LY294002, an inhibitor of phosphatidylinositol 3-kinase. Treatment with LY294002 or knockdown of beta-catenin by small interfering RNA reduced the growth of Epstein-Barr virus-immortalized B-cell line. Transient transfection of latent membrane protein 1 expression plasmid increased beta-catenin protein expression and beta-catenin-dependent transcription. Latent membrane protein 1 deletions mutants lacking the carboxyl terminal activating region 1 domain failed to enhance beta-catenin protein expression and beta-catenin-dependent transcriptional activity. They also failed to increase phosphorylated AKT expression. Dominant-negative AKT suppressed latent membrane protein 1-induced beta-catenin-dependent transcriptional activity. These results suggest that latent membrane protein 1 activates beta catenin through the phosphatidylinositol 3-kinase/AKT signaling pathway. Activation of the beta-catenin pathway by Epstein-Barr virus may contribute to the lymphoproliferation characteristic of Epstein-Barr virus-infected B-cells. PMID- 19309362 TI - Twenty years of cell-penetrating peptides: from molecular mechanisms to therapeutics. AB - The recent discovery of new potent therapeutic molecules that do not reach the clinic due to poor delivery and low bioavailability have made of delivery a key stone in therapeutic development. Several technologies have been designed to improve cellular uptake of therapeutic molecules, including cell-penetrating peptides (CPPs). CPPs were first discovered based on the potency of several proteins to enter cells. Numerous CPPs have been described so far, which can be grouped into two major classes, the first requiring chemical linkage with the drug for cellular internalization and the second involving formation of stable, non-covalent complexes with drugs. Nowadays, CPPs constitute very promising tools for non-invasive cellular import of cargo and have been successfully applied for in vitro and in vivo delivery of therapeutic molecules varying from small chemical molecule, nucleic acids, proteins, peptides, liposomes and particles. This review will focus on the structure/function and cellular uptake mechanism of CPPs in the general context of drug delivery. We will also highlight the application of peptide carriers for the delivery of therapeutic molecules and provide an update of their clinical evaluation. PMID- 19309364 TI - Crosstalk between PTEN/Akt2 and TGFbeta signaling involving EGF receptor down regulation during the tumor promotion process from the early stage in a rat two stage hepatocarcinogenesis model. AB - The present study investigated the involvement of signaling of phosphatase and tensin homolog deleted on chromosome 10 (PTEN)/protein kinase B (Akt) and transforming growth factor-beta (TGFbeta) as well as receptor tyrosine kinases in the tumor promotion processes in a two-stage hepatocarcinogenesis model using male F344 rats. The cellular localization of related molecules was examined in liver cell foci expressing glutathione S-transferase placental form (GST-P) at the early stage of tumor promotion by fenbendazole (FB), piperonyl butoxide, or thioacetamide. Distribution in the liver cell foci and neoplastic lesions positive for GST-P was also examined at the later stage of FB promotion. In contrast to the initiation-alone cases, subpopulations of GST-P-positive foci induced by promotion for 6 weeks, regardless of the promoting chemicals used, enhanced down-regulation of PTEN and up-regulation of phosphorylated (active) Akt2 and phosphorylated substrate(s) of Akt-kinase activity. Also, up-regulation of TGFbeta receptor I and down-regulation of epidermal growth factor receptor (EGFR) were enhanced in the subpopulation of GST-P-positive foci in all promoted cases. A similar pattern of cellular distribution of these molecules was also observed in the neoplastic lesions at the late stage. These results suggest a crosstalk between Akt2 and TGFbeta signaling that involves a mechanism requiring EGFR down-regulation during the entire tumor promotion process starting from the early stage. In particular, a shift in subcellular localization of phosphorylated substrate(s) of Akt from the cell membrane in liver cell foci to the cytoplasm in carcinomas was observed, suggesting an alteration of the function or activity of the corresponding molecule(s). PMID- 19309365 TI - Psoriasis and streptococci: the natural selection of psoriasis revisited. AB - We have previously postulated that surviving invasive streptococcal infections may have been a factor in psoriasis becoming a common skin disease in some parts of the world. Many of the candidate genes linked to psoriasis are associated with the acquired or innate immune system, which are also important in host defence to invasive streptococcal infections. High rates of positive streptococcal throat swabs among patients with chronic plaque psoriasis suggest that they are efficient at internalizing/carrying beta-haemolytic streptococci. Internalization of streptococci in the throat is dependent upon the transforming growth factor (TGF)-beta/fibronectin/alpha 5 beta 1 integrin pathway. The immune cell Th17 and its related cytokine network are important in mucosal defence, being very effective against extracellular microbes but having little effect on intracellular organisms. The TGF-beta/fibronectin/alpha 5 beta 1 integrin pathway and the Th17 cell network also appear to be operative in psoriasis, animal models of both TGF-beta and alpha 5 beta 1 cutaneous overexpression being associated with characteristic psoriasis lesions. We postulate that some of the genotypic/phenotypic changes in different immunological pathways in psoriasis, including the acquired T-cell response, the innate immune response, the TGF beta/fibronectin/alpha 5 beta 1 integrin pathway and the Th17 cell system, confer protection against mortality during epidemics of invasive streptococcal infections, heightened efficiency in internalizing and allowing carriage of streptococci as well as predisposition to the development of psoriasis. PMID- 19309366 TI - Footprints of the EADV: a meeting report from the 17th Congress of the European Academy of Dermatology and Venereology. AB - The 17th Congress of the European Academy of Dermatology and Venereology took place in Paris on 17-20 September 2008 and brought together nearly 11,000 participants. Various plenary lectures, subspecialty meetings, 'free communications', 'top ten', 'test yourself' and 'junior' sessions, 19 courses and 14 'lunches with the expert', six forums, 27 symposia and 45 workshops were pressed into the 4 days of the meeting. Over 1700 posters were presented and exhibited. The themes of a number of symposiums, workshops and sessions overlapped, offering additional educational opportunities despite a very busy schedule. The meeting was well organized. Aesthetic dermatology comprised a significant part of the meeting. It is impossible to encompass all important presentations and we highlight a few medical pearls presented at the meeting; however, our report is not intended as a substitute for reading the conference proceedings and related references quoted in this article. PMID- 19309367 TI - Intense pulsed light vs. long-pulsed dye laser treatment of telangiectasia after radiotherapy for breast cancer: a randomized split-lesion trial of two different treatments. AB - BACKGROUND: Chronic radiodermatitis is a common sequela of treatment for breast cancer and potentially a psychologically distressing factor for the affected women. OBJECTIVES: To evaluate the efficacy and adverse effects of treatments with a long-pulsed dye laser (LPDL) vs. intense pulsed light (IPL) in a randomized split-lesion trial. METHODS: Thirteen female volunteers with radiodermatitis and Fitzpatrick skin types II-III were included in the study. Subjects received a series of three treatments at 6-week intervals with half lesion LPDL (V-beam Perfecta, 595 nm) and half-lesion IPL (Ellipse Flex); the interventions were randomly assigned to left/right or upper/lower halves. Primary end-points were reduction in telangiectasia, patient satisfaction and preferred treatment. Secondary end-points were pain and adverse effects. Efficacy was registered by blinded photographic evaluations 3 months after the final treatment. RESULTS: Eleven patients completed the study. Telangiectasia cleared with both treatments but the efficacy of LPDL was superior. Blinded photographic evaluations showed median vessel clearances of 90% (LPDL) and 50% (IPL) (P = 0.01). LPDL treatments were associated with lower pain scores than IPL treatments [median visual analogue scale (VAS) score 4.3 and 6.0, respectively, P < 0.01]. Patients were slightly more satisfied with LPDL (median VAS score 8) than IPL treatments (median VAS score 7; P < 0.05) and more preferred LPDL (n = 9) to IPL (n = 2) (P < 0.01). Two patients withdrew from the study because of hypopigmentation of the IPL treated areas, which slowly repigmented within 1 year. CONCLUSIONS: This study was based on two specific laser and IPL devices, and found the LPDL treatment to be advantageous compared with IPL due to superior vessel clearance and less pain. PMID- 19309368 TI - Enhanced expression of the antimicrobial peptide LL-37 in lesional skin of adults with atopic eczema. AB - BACKGROUND: Atopic eczema (AE) is a common multifactorial chronic skin disease associated with a defective skin barrier and increased susceptibility to skin infections. The human cathelicidin LL-37 plays a role in the host defence of skin. Studies have demonstrated deficient expression of LL-37 in skin of AE patients. OBJECTIVES: The aim of this study was to investigate the expression of LL-37 in lesional skin compared with nonlesional skin in patients with different severity of AE, patients with other eczema and healthy subjects. METHODS: Twenty patients with AE, four patients with other eczema and 10 healthy subjects were included. Severity of AE was graded using SCORing of atopic dermatitis (SCORAD). Skin biopsies were taken from lesional and nonlesional skin from all patients and from skin of healthy controls. The levels of LL-37 mRNA were analysed by quantitative reverse transcriptase-polymerase chain reaction. Evaluation of dermal and epidermal protein expression of LL-37 and the degree of inflammation was performed by immunohistochemical stainings. RESULTS: Patients with AE and patients with other eczema had significantly (P < 0.05) higher levels of LL-37 in lesional skin than in nonlesional skin. The expression of LL-37 was not statistically associated to severity of AE valued by SCORAD. Nonlesional skin from patients did not differ from skin of healthy subjects in terms of LL-37 expression. In the presence of epidermal injury or vesicles the LL-37 peptide was always detected. CONCLUSIONS: Patients with AE exhibit enhanced expression of LL 37 in lesional skin compared with nonlesional, suggesting a role of LL-37 in AE that might be associated with the process of re-epithelialization. PMID- 19309369 TI - Nonlinear spectral imaging of human hypertrophic scar based on two-photon excited fluorescence and second-harmonic generation. AB - BACKGROUND: A noninvasive method using microscopy and spectroscopy for analysing the morphology of collagen and elastin and their biochemical variations in skin tissue will enable better understanding of the pathophysiology of hypertrophic scars and facilitate improved clinical management and treatment of this disease. OBJECTIVE: To obtain simultaneously microscopic images and spectra of collagen and elastin fibres in ex vivo skin tissues (normal skin and hypertrophic scar) using a nonlinear spectral imaging method, and to compare the morphological structure and spectral characteristics of collagen and elastin fibres in hypertrophic scar tissues with those of normal skin, to determine whether this approach has potential for in vivo assessment of the pathophysiology of human hypertrophic scars and for monitoring treatment responses as well as for tracking the process of development of hypertrophic scars in clinic. METHODS: Ex vivo human skin specimens obtained from six patients aged from 10 to 50 years old who were undergoing skin plastic surgery were examined. Five patients had hypertrophic scar lesions and one patient had no scar lesion before we obtained his skin specimen. A total of 30 tissue section samples of 30 mum thickness were analysed by the use of a nonlinear spectral imaging system consisting of a femtosecond excitation light source, a high-throughput scanning inverted microscope, and a spectral imaging detection system. The high-contrast and high resolution second harmonic generation (SHG) images of collagen and two-photon excited fluorescence (TPEF) images of elastin fibres in hypertrophic scar tissues and normal skin were acquired using the extracting channel tool of the system. The emission spectra were analysed using the image-guided spectral analysis method. The depth-dependent decay constant of the SHG signal and the image texture characteristics of hypertrophic scar tissue and normal skin were used to quantitatively assess the amount, distribution and orientation of their collagen and elastin components. RESULTS: Our experiments and data analyses demonstrated apparent differences between hypertrophic scar tissue and normal skin in terms of their morphological structure and the spectral characteristics of collagen and elastin fibres. These differences can potentially be used to distinguish hypertrophic scar tissues from normal skin and to evaluate treatment responses. CONCLUSIONS: All the measurements were performed in backscattering geometry and demonstrated that nonlinear spectral imaging has the ability to differentiate hypertrophic scar tissue from normal skin based on noninvasive SHG imaging, and TPEF imaging revealed the microstructure and spectral features of collagen and elastin fibres. With the advances in spectral imaging apparatus miniaturization, we have good reason to believe that this approach can become a valuable tool for the in vivo pathophysiology study of human skin hypertrophic scars and for assessing the treatment responses of this disfiguring disease in clinic. It can also be used to track the development of hypertrophic scars and to study wound healing processes in a noninvasive fashion without biopsy, fixation, sectioning and the use of exogenous dyes or stains. PMID- 19309370 TI - Pruritus measurement and treatment. AB - Pruritus measurement is problematic, because of its subjective nature and poor localization. Ratio scales enhance the usefulness of the visual analogue scale (VAS) by reducing variation; other scales such as the generalized labelled magnitude scale may also be useful. Pruritus neuroanatomy includes peripheral receptors, peripheral and central nerves, ascending and descending spinal pathways, and several brain regions. Pruritus receptors include Merkel discs and free nerve endings, and itch receptors have fast or slow adaptation. In this review, we discuss the pathophysiology of pruritus in atopic dermatitis, psoriasis and scabies. Pruritus treatment is reviewed for topical agents and antihistamines. Future research directions are suggested. PMID- 19309371 TI - Oral ulcers: clinical aspects. A tool for dermatologists. Part I. Acute ulcers. AB - Oral ulcers are generally painful lesions that are related to various conditions developing within the oral cavity. They can be classified as acute or chronic according to their presentation and progression. Acute oral ulcers are be associated with conditions such as trauma, recurrent aphthous stomatitis, Behcet's disease, bacterial and viral infections, allergic reactions or adverse drug reactions. Chronic oral ulcers are associated with conditions such as oral lichen planus, pemphigus vulgaris, mucosal pemphigoid, lupus erythematosus, mycosis and some bacterial and parasitic diseases. The correct differential diagnosis is necessary to establish the appropriate treatment, taking into account all the possible causes of ulcers in the oral cavity. In the first part of this two-part review, acute oral ulcers are reviewed. PMID- 19309372 TI - Pityriasis rubra pilaris type 1 spontaneously resolving after 20 years. AB - Pityriasis rubra pilaris (PRP) is an uncommon, idiopathic, papulosquamous eruption. We report a longitudinal study of a patient with PRP type 1 who was treated with retinoid therapy for 9 years and whose symptoms resolved spontaneously after 20 years. There are no data in the literature on the disease course of PRP type 1 persisting beyond the usual 3 years. This case highlights both the extreme chronicity of PRP and the possibility of remission after many years of active disease. PMID- 19309373 TI - Increased caspase-3 and altered expression of apoptosis-associated proteins, Bcl 2 and Bax in lichen planus. AB - BACKGROUND: Lichen planus (LP) is a chronic inflammatory disease of probable immune-based aetiology. The pathogenesis of LP is unclear, but apoptotic changes in epidermal (epithelial) cells have been reported. OBJECTIVE: To evaluate apoptosis in LP through studying caspase-3 expression and to determine whether the apoptosis-associated proteins Bcl-2 and Bax are significantly involved in the pathogenesis of LP. METHODS: In total, 25 lesional biopsy specimens [15 cutaneous LP (CLP) and 10 oral LP (OLP)] and 10 control specimens [5 normal skin and 5 normal oral mucosa] were studied using immunochemical methods for the expression of caspase-3, Bcl-2 and Bax proteins. RESULTS: Compared with controls, a significant increase in caspase-3 and Bax protein expressions were found in LP lesions. Basal cell expression of caspase-3 was positive in 14 cases (56%), and 12 cases (48%) showed mild expression. Caspase-3 expression in inflammatory infiltrate was positive in 13 cases (52%). Of these, 12 cases (48%) showed mild positivity. Bax was localized mostly to the upper prickle layer. Basal cell expression of Bcl-2 was negative in 18 (72%) cases, with no significant difference between patients with LP and controls. Bcl-2 was expressed in the inflammatory infiltrate in 15 cases of LP (60%), showing mild expression in 12 cases (48%). Compared with CLP, there was a significant increase in caspase-3 expression in OLP, despite the nonsignificant difference in Bcl-2 and Bax protein expressions by the epithelial cells. CONCLUSION: Increased caspase-3 and altered expression of Bcl-2 and Bax were found in LP, indicating the possible involvement of these proteins in the pathogenesis of the disease. The observed increase in apoptosis in OLP compared with CLP might explain the difference in clinical behaviour that distinguishes these LP variants. PMID- 19309374 TI - Expression and localization of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 in human epidermal appendages: a comparison study by immunofluorescence. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) promotes angiogenesis and plays important roles in neovascularization and development of tissues. VEGF receptors (VEGFRs) are high-affinity receptors for VEGF and are originally considered specific to endothelial cells. We have previously shown that keratinocytes from human normal skin express VEGFRs. This poses the question of whether these receptors are also expressed by epidermal appendages, as epidermal appendages are lined with epithelial cells. OBJECTIVE: To investigate the expression of VEGFR-2 compare with VEGF in epidermal appendages, including hair follicles, eccrine sweat glands and sebaceous glands. METHODS: Monoclonal antibodies to VEGF and VEGFR-2 were used for immunohistochemical examination of cryostat-cut sections of normal human skin specimens from 11 donors undergoing cosmetic surgery. RESULTS: Immunoreactivities for VEGF and VEGFR-2 principally showed parallel intense expression in anagen hair follicle (including outer root sheat, inner root sheath, dermal papillae epidermal matrix), sebaceous glands (ductal and secretory portions) and eccrine sweat glands (ductal and secretory portions), respectively. In particular, abundant expression of VEGF was found in the follicular basement membrane zone surrounding the bulb matrix and in the ductal and secretory portions of eccrine sweat glands. CONCLUSION: A potential VEGF/VEGFR-2 autocrine pathway may be defined by the coexpression of VEGF and VEGFR-2 in human skin epidermal appendages. PMID- 19309375 TI - Autoimmune hepatitis associated with infliximab in a patient with palmoplantar pustular psoriaisis. PMID- 19309376 TI - Bilateral naevus of Ota in a pregnant white woman. PMID- 19309377 TI - The axial frontonasal flap for reconstruction of large nasal-tip defects based on modified nasal subunits in East Asians. AB - The nasal tip is a prominent landmark on the face, and skin carcinomas of this area are very common. Moderate and large deep skin defects of the nasal tip normally represent a difficult reconstructive challenge. Nasal-tip reconstruction techniques have evolved to allow not only filling of the gap, but cosmetic and functional retention. Numerous flap options are available to reconstruct the defect of the tip of the nose. In this paper, we report the technique of the axial frontonasal flap and its use in reconstruction of large nasal-tip defects in East Asians. The axial frontonasal flap described is a good alternative for reconstruction of nasal-tip defects > 15 mm in diameter, with excellent aesthetic results. This technique is reliable, yields excellent functional and cosmetic results, and we believe that it is a valuable alternative to other techniques of reconstruction for defects of the nasal tip. PMID- 19309378 TI - An ulcerated plaque on the hard palate. PMID- 19309379 TI - A unilateral periorbital papulonodular eruption. PMID- 19309380 TI - Multiple asymptomatic scrotal nodules. PMID- 19309381 TI - Familial yellow papules. PMID- 19309382 TI - An auricle nodule. PMID- 19309383 TI - A progressive eruption in a 57-year-old woman. PMID- 19309384 TI - Xeroderma pigmentosum: early diagnostic features and an adverse consequence of photoprotection. PMID- 19309385 TI - Pyoderma gangrenosum following pacemaker insertion. PMID- 19309386 TI - An (un)lucky blow to the nail? PMID- 19309387 TI - Fatal muscle haemorrhage attributable to heparin administration in a patient with dermatomyositis. PMID- 19309389 TI - Influence of gastric mucosal status on success of stepwise acid suppressive therapy for dyspepsia. AB - BACKGROUND: The most effective initial treatment strategy of dyspepsia is still under debate. Individual biological characteristics, such as condition of gastric mucosa, might contribute to selection of the most appropriate acid suppression treatment strategy. AIM: To assess whether pre-treatment testing of gastric mucosal status is relevant for treatment success in an RCT comparing step-up and step-down therapies in newly diagnosed dyspepsia patients. METHODS: Baseline serum samples were collected to assess gastric mucosal status using serum levels of pepsinogens-I&II, gastrin-17, and Helicobacter pylori IgA/IgG-antibodies. The 6-month treatment success was compared between step-up and step-down for patients with serum diagnoses: normal; gastritis; corpus atrophy or antrum atrophy. RESULTS: In all, 519 patients (M/F: 249/270, age: 47 (18-85) years, 29%H. pylori+) were randomized to step-up (n = 293) or step-down (n = 226). Normal mucosa, gastritis and corpus atrophy were diagnosed serologically in 70%, 28% and 2% of the patients, evenly distributed between the strategies (P = 0.65). Treatment success was achieved in respectively, 69%, 70% and 70% for the serum diagnosis groups, and did not differ between the strategies. CONCLUSIONS: Dyspepsia treatment success could not be predicted by gastric mucosal status. Therefore, serum diagnosis of gastric mucosal status is no useful tool for patient allocation to acid suppressive treatment strategies. PMID- 19309390 TI - NSAID-induced antral ulcers are associated with distinct changes in mucosal gene expression. AB - BACKGROUND: The basis for individual variation in gastroduodenal vulnerability to NSAIDs is not well understood. AIM: To assess whether a gene expression signature is associated with susceptibility to gastroduodenal ulcerations. METHODS: Twenty five Helicobacter pylori negative adults were treated for 7 days with naproxen 500 mg b.d. Subjects underwent baseline and post-treatment endoscopy, during which biopsies were taken from antrum and duodenum. RNA extraction and cDNA synthesis were performed, followed by PCR of 23 genes relevant to mucosal injury and repair. Fold changes in gene expression were compared between subjects who developed ulcers and those who did not. RESULTS: Compared with subjects who did not develop ulcers (n = 18), subjects who developed antral ulcers (n = 7) had significantly greater mucosal up-regulation of interleukin-8 [Fold change = 33.5 (S.E.M. = 18.5) vs. -7.7 (3.2)] and of cyclo-oxygenase-2 [2.3 (1.7) vs. -10.8 (2.2)]. Conversely, non-ulcer subjects had significantly greater up-regulation of toll-like receptor-4, cyclo-oxygenase-1 and hepatocyte growth factor [14.0 (2.2) vs. -0.8 (1.0), 9.8 (2.4) vs. 0.0 (0.7) and 8.2 (2.6) vs. -2.2 (0.3) respectively]. CONCLUSIONS: NSAID-induced antral ulcers are associated with a specific pattern of gastroduodenal mucosal gene expression. These patterns may provide an insight into the molecular basis of individual susceptibility to mucosal injury. PMID- 19309391 TI - Effects of repeated injections of fibroblast-stimulating lipopeptide-1 on fever, formation of cytokines, and on the responsiveness to endotoxin in guinea-pigs. AB - AIMS: We investigated, whether the Toll-like receptors (TLRs)-2/6-agonist fibroblast-stimulating lipopeptide-1 (FSL-1), like the TLR-4 agonist lipopolysaccharide (LPS), induces a state of tolerance. We further tested the influence of repeated pre-treatment with FSL-1 on the animals' responsiveness to LPS. METHODS: Abdominal temperature was recorded in unrestrained guinea-pigs with intra-abdominally implanted radiotransmitters. Circulating concentrations of tumour necrosis factor (TNF) and interleukin-6 (IL-6) were measured with specific bioassays. We tested the effects of intra-arterial (i.a.) or intraperitoneal (i.p.) injections of 100 microg kg(-1) FSL-1, repeated five times at intervals of 3 days. The animals' responses to i.a. or i.p. injections of 10 microg kg(-1) LPS were determined another 3 days later and compared to those of naive guinea-pigs. RESULTS: The FSL-1-induced TNF peak was significantly attenuated starting with the third i.a. administration, while fever was unimpaired and the IL-6-peak just tended to decrease. Fever and IL-6 in response to i.a. injections of LPS were identical in both groups, while circulating TNF was higher in naive compared to FSL-1 pre-treated animals. The effects of repeated i.p. injections of FSL-1 were more pronounced resulting in attenuation of fever as well as circulating TNF and IL-6, the strongest reduction observed after the third stimulation with FSL-1. Repeated i.p. pre-treatment with FSL-1 induced hyporesponsiveness to i.p. administration of LPS compared to naive animals with regard to fever and especially with regard to LPS-induced formation of cytokines. CONCLUSIONS: There is a development of tolerance to FSL-1 and cross-tolerance between FSL-1 and LPS depending on the route of administration of the respective TLR-2/6 and TLR-4 agonists. PMID- 19309393 TI - The expression of ERalpha, ERbeta and PR in lobular carcinoma in situ of the breast determined using laser microdissection and real-time PCR. AB - AIMS: To determine the status of oestrogen receptor (ER) subtypes (ERalpha and ERbeta) in lobular carcinoma in situ (LCIS) of the breast. METHODS AND RESULTS: Forty-seven cases of LCIS and six cases with normal breast lobules were subjected to immunohistochemistry and evaluated for ERalpha, ERbeta and progesterone receptor (PR) expression. mRNA for ERalpha, ERbeta1 and ERbeta2 were quantified in LCIS and normal lobules using laser microdissection coupled with real-time polymerase chain reaction. LCIS showed a higher level of steroid receptor protein expression than normal lobules. There was no difference in ERbeta1 gene or ERbeta protein expression between normal lobules, pure LCIS, or LCIS associated with invasive breast cancer. No significant difference in expression of either ERalpha or ERbeta was found between pure LCIS and LCIS associated with invasive cancer. However, PR was significantly lower in those cases of LCIS with associated invasive than in those without synchronous invasive disease. CONCLUSIONS: Increased expression of steroid receptors in LCIS suggests their possible role in the biology of LCIS and, for PR, could influence the predisposition of women diagnosed with LCIS to develop invasive breast carcinoma. PMID- 19309394 TI - Rapid screening of tissue microarrays for Her-2 fluorescence in situ hybridization testing is an accurate, efficient and economic method of providing an entirely in situ hybridization-based Her-2 testing service. AB - AIMS: Fluorescence in situ hybridization (FISH) testing is the 'gold standard' method for Her-2 status assessment in breast cancer patients, yet is only employed in about 30% of tests carried out because of cost and labour considerations. We have previously described tissue microarray (TMA)-based testing to eliminate cost constraints, and now describe a rapid screening approach to reduce time spent testing. METHODS AND RESULTS: We examined 88 cases of invasive breast cancer on TMAs comparing formal FISH scoring with a rapid screening technique. Each core was screened by two observers and results recorded as positive, equivocal or negative. Each approach was timed. Data were analysed by comparing the rapid screening results with formal counts. Using rapid screening, two-thirds of negative and half the positive cases could be eliminated with 100% accuracy. It took 2 min per observer per case to rapid screen six TMA cores at x100 magnification. The remaining cases required formal counting, which took no longer than with whole-section techniques. CONCLUSION: Rapid screening of TMAs for routine Her-2 FISH testing is safe, economical and time efficient. The technique ensures that all patients receive 'gold standard' testing. PMID- 19309395 TI - Methylene blue-assisted lymph node dissection in combination with ex vivo sentinel lymph node mapping in gastric cancer. AB - AIMS: Lymph node (LN) stage is still the strongest prognostic marker in potentially curable gastric cancer. Accuracy of histopathological lymph node assessment depends on the number of investigated LNs and detection rate of metastases and micrometastases. The aim was to perform a feasibility study employing intra-arterial methylene blue injection - a novel method to improve LN harvest - and ex vivo sentinel LN mapping. METHODS AND RESULTS: A total of 33 cases were enrolled, including 14 retrospective cases that served as a control group. The methylene group showed a highly significant improved mean LN harvest compared with unstained cases, with 38 +/- 14 versus 21 +/- 10 LNs (P < 0.001), respectively. The detection rate of ex vivo sentinel mapping was 88%. No skip metastases occurred. CONCLUSION: Both techniques have the potential to improve the accuracy of histopathological LN staging and can be combined successfully. PMID- 19309396 TI - Prognostic importance of epithelial-mesenchymal transition-related protein expression in gastric carcinoma. AB - AIMS: Epithelial-mesenchymal transition (EMT) is defined as switching of polarized epithelial cells to a migratory fibroblastoid phenotype. EMT is known to be involved in the progression and metastasis of various cancers. The aim was to evaluate the expression of EMT-related proteins in gastric carcinoma (GC). METHODS AND RESULTS: The expression of nine EMT-related proteins in the GC tissues of 598 patients was evaluated by immunohistochemistry using a tissue array method. In addition, clinicopathological characteristics and survival were compared with the expression of EMT-related proteins. Loss of epithelial protein and/or acquisition of the expression of mesenchymal proteins were observed in GC. These protein alterations were associated with diffuse type histology, advanced stage and poor patient outcome, respectively. Subjects were divided into three groups according to degree of EMT-related protein alteration. Increases in alteration of EMT-related protein were found to be significantly associated with poorly differentiated histology, higher pTNM stage and unfavourable outcome. Multivariate analysis showed that alterations in the expression of EMT-related proteins were independently associated with poor prognosis. CONCLUSIONS: Loss of epithelial proteins and/or the acquisition of mesenchymal proteins are associated with poorly differentiated histology, advanced stage and poor outcome in GC. The awareness and inhibition of EMT offer a promising target for prevention of metastatic progression and invasion. PMID- 19309397 TI - Decreased intratumoral arteries reflect portal tract destruction and aggressive characteristics in intrahepatic cholangiocarcinoma. AB - AIMS: To assess the relationship between arterial blood supply and the progression of intrahepatic cholangiocarcinoma (ICC). METHODS AND RESULTS: The intratumoral arterial vessel density (AVD) was assessed in 76 cases of mass forming type of ICC using anti-h-caldesmon antibody, a marker of smooth muscle cells, and AVD compared with pathological findings. AVD was directly correlated with the presence of intratumoral portal tracts (P < 0.0001) and inversely correlated with the grade of tumour necrosis (P = 0.0013). AVD was inversely correlated with vascular invasion and lymph node metastasis (P = 0.0159 and P = 0.0023, respectively). The hilar type of ICC had lower AVD regardless of tumour size, whereas the peripheral type with high AVD showed branching ductular formation composed of cuboidal cells with mild nuclear atypia. AVD was found to be an independent prognostic factor on multivariate survival analysis (P = 0.0013). CONCLUSIONS: This study demonstrates that intratumoral arterial vessels reflect engulfed portal tracts in ICC and decreased arterial vessels indicate aggressive tumour behaviour. Our results could contribute to clinical tumour staging and more effective therapy. PMID- 19309398 TI - Expression of glucose-regulated stress protein GRP78 is related to progression of melanoma. AB - AIMS: Glucose-regulated protein 78 (GRP78) is a protein translated in response to endoplasmic reticulum (ER) stress that has been implicated in the pathogenesis and resistance to therapy of a variety of cancers. The aim of this study was to investigate its expression and role in the development and progression of human melanoma. METHODS AND RESULTS: The immunohistochemical expression of GRP78 in naevi, primary melanoma and melanoma metastases from 171 patients was correlated with clinicopathological factors and patient survival. The GRP78 immunoreactivity score (IRS) was 0.2 in compound naevi, 0.65 in dysplastic naevi, 4.65 in naevi adjacent to primary melanoma, 2.4 in melanoma in situ, 11.2 in thin (1.0 mm) primary melanoma. It was 18 and 17.3 in subcutaneous and lymph node metastases, respectively (P < 0.0001). GRP78 expression was positively correlated with increasing tumour thickness (P = 0.001) and with increasing dermal tumour mitotic index (P = 0.0004). Disease-free survival (chi(2) = 8.0703, P = 0.0045) and overall survival (chi(2) = 6.2633, P = 0.0123) in melanoma patients with IRS >25 were significantly lower than in melanoma patients with IRS <25. CONCLUSIONS: GRP78 expression appears to correlate with known correlates of melanoma progression and survival and requires further evaluation as a prognostic biomarker in melanoma. PMID- 19309399 TI - Retiform uterine tumours resembling ovarian sex cord tumours. A comparative immunohistochemical study with retiform structures of the female genital tract. AB - AIMS: To analyse both the clinicopathological and immunohistochemical findings in six cases of uterine tumours resembling ovarian sex cord tumours with a predominant epithelial retiform component resembling the rete ovarii (RUTROSCT) and to compare their immunophenotype with tissues containing retiform structures such as the normal rete ovarii, retiform Wolffian adnexal tumour (RWAT) and ovarian retiform areas of Sertoli-Leydig cell tumours (ORSLCT). METHODS AND RESULTS: Six RUTROSCTs were analysed. The average age of patients was 65 years, and all tumours behaved in a benign fashion. Four were intracavitary and two intramural. Two cases were misdiagnosed as malignant in endometrial biopsy specimens and one in the hysterectomy specimen. Histologically they had a tubulopapillary or glomeruloid formation; a sex-cord-like or endometrial stromal component was absent or comprised at most 30% of the tumour. RUTROCST showed consistent positivity for CAM5.2, cytokeratin (CK) 7, vimentin, calretinin, progesterone receptor and apical CD10. CD56 and alpha-inhibin were positive in half of the cases. Epithelial membrane antigen, desmin, smooth muscle actin and calponin were negative. Comparatively, rete ovarii and RWAT had a similar positivity for CK7, CD56, CD10 and calretinin. ORSLCT differed in its conspicuous positivity to CD56, CD10, alpha-inhibin and calretinin, but absent CK7 expression. CONCLUSIONS: RUTROSCTs have benign behaviour but may be confused with various uterine adenocarcinomas or metastases. Correct diagnosis should avoid overtreatment. The immunophenotype of retiform areas is similar to that of adult rete ovarii and RWAT with minor divergence from ORSLCT. PMID- 19309400 TI - Bcl-10 protein highly correlates with the expression of phosphorylated p65 NF kappaB in peripheral T-cell lymphomas and is associated with clinical outcome. AB - AIMS: In T cells, protein kinase C (PKC) theta plays a major role in T-cell receptor (TCR)-mediated activation of a novel nuclear factor (NF)-kappaB pathway that involves phosphorylation of p65 at serine 536 (Pp65(Ser536)). Bcl-10 acts along the same pathway downstream of PKC theta to activate NF-kappaB. The aim was to investigate the relationship between the expression of PKC theta, Bcl-10 and P p65(Ser536) proteins and their prognostic significance in peripheral T-cell lymphomas (PTCLs). METHODS AND RESULTS: Paraffin-embedded tissues from 30 patients with PTCLs treated with curative intention were evaluated retrospectively. Expression of PKC theta, Bcl-10 and P-p65(Ser536) proteins was assessed using immunohistochemistry. Expression of PKC theta was detected in 22 of 30 cases (73%), Bcl-10 in 20 of 30 (67%) and P-p65(Ser536) in 21 of 30 (70%). Bcl-10+ tumours were associated with PKC theta (18 of 22) (P < 0.0001) and Pp65Ser536 (19 of 21) expression (P < 0.0001). Patients with Bcl-10+ or P p65(Ser536+) tumours fared better, with a 5-year overall survival of 48 and 45%, respectively, versus 0% for negative tumours (P = 0.029 and P = 0.04, respectively). CONCLUSIONS: Bcl-10 is expressed in PTCLs, correlates with PKC theta and Pp65(Ser536) expression and seems to be associated with better survival. PMID- 19309401 TI - Bone marrow necrosis related to paracoccidioidomycosis: the first eight cases identified at autopsy. AB - AIMS: To report the first eight bone marrow necrosis (BMN) cases related to paracoccidioidomycosis (PCM) from patient autopsies with well-documented bone marrow (BM) histology and cytology. METHODS AND RESULTS: A retrospective evaluation was performed on BM specimens from eight autopsied patients from Botucatu University Hospital with PCM-related BMN. Relevant BMN literature was searched and analysed. CONCLUSIONS: All eight patients had acute PCM. Six had histological only (biopsies) and two cytological only (smears) specimens. Five biopsy specimens revealed severe and one mild coagulation patterned necrotic areas. Five had osteonecrosis. The cytological specimens also showed typical BMN patterns. Paracoccidioides brasiliensis yeast forms were visible within necrotic areas in all cases. PMID- 19309402 TI - Inflammatory myofibroblastic tumour of the urinary bladder mimicking recurrent uterine carcinosarcoma. PMID- 19309404 TI - Is sclerosing angiomatoid nodular transformation (SANT) of the splenic red pulp identical to inflammatory pseudotumor? Report of 16 cases. PMID- 19309405 TI - Tubal and ovarian pathways to pelvic epithelial cancer: a pathological perspective. PMID- 19309406 TI - Interpretation of diffuse Cdx2 expression in endometrioid adenocarcinoma in the absence of morules. PMID- 19309407 TI - Uterine and extrauterine plexiform tumourlets are sex-cord-like tumours with myoid features. PMID- 19309408 TI - Metastasizing anaplastic ependymoma in an adult. Chromosomal imbalances, metabolic and gene expression profiles. PMID- 19309409 TI - MAP-2 immunoexpression in gliomatosis cerebri. PMID- 19309410 TI - Atypical organizing haematoma/seroma following treatment for soft tissue sarcoma. PMID- 19309411 TI - Activation of NF-kappaB transcription factor in asthma death. PMID- 19309412 TI - The expression of activation induced cytidine deaminase in follicular lymphoma is independent of prognosis and stage. PMID- 19309413 TI - The story of O--is oxytocin the mediator of the placebo response? AB - While the placebo responses in various medical conditions have been shown to follow a few basic principles such as expectancies, reward learning and Pavlovian conditioning, the underlying neurobiology and the mediating hormonal and/or neuromodulating processing have remained obscure. We here report the collected evidence that oxytocin (OXT), a 389-amino acid polypeptide located on chromosome 3p25 that is released in the brain (hypothalamus) and in peripheral tissue, is the central mediator of the placebo response: we hypothesize that exogenous OXT via an OXT agonist will enhance the placebo response, while exogenous OXT blockade by an antagonist will reduce the placebo response in placebo analgesia and other placebo models. It is furthermore proposed that the placebo response in trials may be predicted by circulating plasma OXT levels, the OXT receptor density in the brain and/or the presence of one or more of the single nucleotide polymorphisms of the OXT promoter gene. PMID- 19309414 TI - The challenge of cross-cultural, multi-national research: potential benefits in the functional gastrointestinal disorders. AB - The increasing interest in research in irritable bowel syndrome (IBS) and other functional gastrointestinal disorders (FGIDs), taken together with the growing sophistication of communication technology, makes cross-cultural, multi-national research a feasible endeavour. The aim of this study is to encourage collaborative cross-cultural studies in FGIDs by discussing relevant methodological issues, and by suggesting potential areas in which cross-cultural research can make a significant contribution to the understanding of FGIDs and to patient care. To this end, methodological issues related to cross-cultural research and competences required for its conduct are presented together with a critique of published studies and recommendations for future research in the area. The term 'cross-cultural' research in FGIDs is usually applied to the results of prevalence studies, for example comparative studies of IBS prevalence in different countries and ethnic groups. The validity of these comparisons is impacted negatively by the lack of uniformity in research methods. In addition to prevalence studies, cross-cultural research can make a significant contribution in areas such as molecular biology, genetics, psychosocial factors, symptom presentation, extra-intestinal comorbidity, diagnosis and treatment, determinants of disease severity, healthcare utilization, and health-related quality of life, all issues that can be affected by culture, ethnicity and race. Well-designed and implemented cross-cultural studies can advance our knowledge in many FGID-related areas ranging from epidemiology through psychosocial factors, pathophysiological mechanisms and therapeutics. These studies, conducted by investigators with competence in cross-cultural research methodology, can advance our understanding of the FGIDs and contribute to improved patient care. PMID- 19309415 TI - Pharmacogenetics of low dose clonidine in irritable bowel syndrome. AB - Adrenergic and serotonergic (ADR-SER) mechanisms alter gut (gastrointestinal, GI) sensorimotor functions. We aimed to determine whether candidate ADR-SER genes affect GI responses to low dose clonidine (CLO) in humans. Forty healthy and 120 irritable bowel syndrome (IBS) participants received CLO, 0.1 mg or 0.15 mg b.i.d., for 6 days. At baseline and post-CLO, we measured: gastric volume (GV); satiation volume; rectal compliance, sensation thresholds and ratings with distensions. Genetic variations tested were: alpha2A (C-1291G), alpha2C (Del 322 325), GNbeta3 (C825T) and solute carrier family 6 (neurotransmitter transporter, serotonin), member 4 (SLC6A4) (serotonin transporter linked polymorphic region). CLO reduced volume to satiation (P = 0.002), postprandial GV (P < 0.001), sensation threshold for pain (<0.001); CLO increased rectal compliance (P = 0.024). There were significant associations between post-CLO responses and gene variations for DeltaGV (alpha2A and SLC6A4), rectal sensation of gas (alpha2A, GNbeta3), urgency (alpha2A); and pain (GNbeta3 and SLC6A4); and rectal compliance (SLC6A4). alpha2A, GNbeta3 and SLC6A4 genotypes significantly modify responses to CLO on sensory and motor GI functions in health and IBS. PMID- 19309416 TI - Oxytocin receptor expressed on the smooth muscle mediates the excitatory effect of oxytocin on gastric motility in rats. AB - The aim of this study was to localize oxytocin receptor (OTR) in the stomach and to investigate the effect of OT on gastric motility in rats. Western blot and immunohistochemistry methods were used to localize OTR in stomach. The motility of stomach was recorded in vivo (recording of the intragastric pressure), in vitro (recording of the contraction of muscle strips) and on isolated smooth muscle cells. OTR was expressed on cells of both circular and longitudinal muscle of stomach. Systemic administration of OT induced an early transient decrease and a subsequent increase on intragastric pressure. Devazepide (1 mg kg(-1), i.v.), a cholecystokinin-1 (CCK(1)) receptor antagonist, completely abolished the transient response but did not influence the subsequent one. OT (10(-9)-10(-6) mol L(-1)) dose-dependently increased the contraction of the muscle strips of gastric body, antrum, and pyloric sphincter, and decreased the average cell length of isolated smooth muscle cells. Tetrodotoxin and atropine did not influence the effect of OT on muscle strips. Pretreatment with atosiban, an OTR antagonist, inhibited the spontaneous contraction of muscle strips and abolished the excitatory effect of OT on the muscle strips and the isolated cells. These results suggest that the OTR is expressed on the smooth muscle of the stomach and mediates excitatory effect of OT on gastric motility. PMID- 19309417 TI - Myxobolus erythrophthalmi sp. n. and Myxobolus shaharomae sp. n. (Myxozoa: Myxobolidae) from the internal organs of rudd, Scardinius erythrophthalmus (L.), and bleak, Alburnus alburnus (L.). AB - During a survey of myxosporean parasites of cyprinid fish in Hungary, infections caused by unknown Myxobolus spp. were found in the internal organs of rudd, Scardinius erythrophthalmus, and bleak, Alburnus alburnus. Small plasmodia developed in blood vessels of the kidney, liver, testes and intestinal wall. The parasites were studied on the basis of spore morphology and by histological and molecular methods. In most cases, plasmodia were surrounded by host tissue without a host reaction; however, in advanced cases, a connective tissue capsule was seen around plasmodia. Spores collected from the two fish species differed from each other and from the known Myxobolus spp. both in their morphology and 18S rDNA sequences. The two species, described as M. erythrophthalmi sp. n. from rudd and M. shaharomae sp. n. from bleak, are characterized by a specific histotropism to blood vessels, while the organ specificity involves the kidney and for the latter species, most internal organs. PMID- 19309418 TI - Changes in vertebral structure during growth of reared rainbow trout, Oncorhynchus mykiss (Walbaum): a new approach using modelling of vertebral bone profiles. AB - Severe bone resorption of the vertebral body in reared rainbow trout was thought to be a dysfunction in mineral balance induced by increased growth rate in unfavourable rearing conditions. To verify this assumption, we sampled market sized trout (c. 250 g) from 20 fish farms with different rearing conditions. Growth rate was also studied by sampling trout reared in three different water temperatures from fry to market-size. Transverse sections of vertebrae were microradiographed, then digitized. Total bone area (Tt-B.Ar.) and bone profiles were obtained using BONE PROFILER 3.23 software and a mathematical model was developed to statistically compare bone profiles using 12 parameters in four vertebra regions. Tt-B.Ar. and bone profiles were found to vary with rearing conditions and growing temperatures, indicating obvious influences of these factors on bone remodelling. However, vertebral resorption was found to be a general phenomenon. In trout from 190 to 235 mm in length, vertebrae underwent important remodelling resulting in large resorption of the middle area, while the transition and peripheral areas showed an increase in bone deposition. Changes in vertebra architecture seem to be a good compromise between the need to mobilize stored minerals during growth while maintaining vertebral biomechanical properties. PMID- 19309419 TI - Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms. AB - Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus, caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10(7) copies microg(-1) DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10(6)-10(8) copies of CyHV-2 microg(-1) DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22-10 degrees C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (+/-SE) of 7.3 +/- 11 to 394 +/- 55 microg(-1) DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease. PMID- 19309420 TI - The effects of a home-based intervention for young children with intellectual disabilities in Vietnam. AB - BACKGROUND: This study was conducted to examine the impact of a 1-year intervention for children with intellectual disabilities (ID) in Vietnam. METHOD: Subjects were 30 preschool-aged children with ID (ages 3 to 6 years). Sixteen were assigned to an intervention group and 14 to a control group. Based on the Portage Curriculum (CESA 5 2003), the intervention trained parents to work with their children through modelling and coaching by teachers during weekly home visits. RESULTS: Comparison of pre-, mid- and post-intervention assessments of the children based on the Vineland Adaptive Behavior Scales (Sparrow et al. 1984a) indicated that the intervention was promising: children in the intervention group improved significantly in most domains of adaptive behaviours, and also performed significantly better than the control group in the areas of personal care and motor skills. CONCLUSIONS: The results from the Vietnam programme are discussed in terms of its implications and strategies for developing programmes for children with disabilities in developing countries. PMID- 19309421 TI - Prolactin levels in the patients with vitiligo. PMID- 19309422 TI - Pimecrolimus: a new choice in the treatment of vitiligo? PMID- 19309423 TI - Onychomycosis and diabetes. AB - OBJECTIVE: This study aims to discuss factors specific to diabetics in the diagnosis and treatment of onychomycosis. DISCUSSION: Onychomycosis has the potential to cause severe complications in diabetics and should be treated promptly. The existence of comorbid conditions and potential for drug-drug interactions complicates the selection of an appropriate treatment regimen. The role of Candida in onychomycosis is controversial but may be of increased significance in the diabetic population due to an underlying vulnerability to this organism. CONCLUSIONS: Terbinafine is an excellent choice in diabetics due to its low risk of drug-drug interaction and proven efficacy against the typical pathogens that cause onychomycosis. Itraconazole, while an effective treatment for onychomycosis, is not a first-choice therapy due to its black-box cardiac warning and numerous drug interactions. Larger studies are needed in diabetics to determine the frequency of candidal nail infections. PMID- 19309424 TI - Fractional ablative CO(2) laser resurfacing improves a thermal burn scar. PMID- 19309425 TI - The use of dermoscopy for the diagnosis of trichotillomania. PMID- 19309426 TI - The use of dermoscopy for the diagnosis of plantar wart. PMID- 19309427 TI - Vascular endothelial growth factor in psoriasis: an indicator of disease severity and control. AB - BACKGROUND: Psoriasis is a chronic disease characterized by abnormal epidermal proliferation, inflammation and angiogenesis. It has been reported that vascular endothelial growth factor (VEGF) is overexpressed in lesional psoriatic skin and its serum levels are significantly elevated in patients with moderate to severe disease. OBJECTIVE: This study aims to evaluate the possible role of VEGF in the pathogenesis of psoriasis, and its significance as an indicator of disease severity and control. METHODS: Thirty patients with moderate to severe psoriasis and 10 healthy controls were subjected to baseline evaluation of VEGF. Patients were divided into three groups according to the received treatment: psoralen plus ultraviolet A (PUVA) thrice weekly (group 1), acitretin 50 mg daily (group 2), and combined PUVA twice weekly and acitretin 25 mg daily (group 3).Treatment continued for 16 weeks or up to clinical cure. Every patient was subjected to severity evaluation by Psoriasis Area and Severity Index (PASI) and measurement of serum VEGF before and after treatment. RESULTS: Mean serum levels of VEGF were significantly elevated in patients (327 +/- 66.2 pg/mL) than control subjects (178 +/- 83.4 pg/mL). A highly significant correlation was found between VEGF and PASI score, but not with other variables. The best clinical response, the least side-effects and the highest reduction of VEGF serum levels were achieved by the combined therapy. CONCLUSION: The present study supported the proposed role of VEGF in the pathogenesis of psoriasis, and suggested that it could serve as a good indicator of disease severity and control. PMID- 19309428 TI - Variable response of crusted scabies to oral ivermectin: report on eight Egyptian patients. AB - BACKGROUND: Several reports have proved the efficacy of oral ivermectin in the treatment of crusted scabies. However, the response varied greatly between different studies. OBJECTIVE: The aim of this study was to evaluate the response of crusted scabies to oral ivermectin in eight Egyptian patients. PATIENTS AND METHODS: Eight patients with crusted scabies, diagnosed clinically and confirmed microscopically, were involved in this study. Patients received a single oral dose of ivermectin (200 microg/kg) and re-examined at 2, 4, 6 and 8 weeks. A second dose of ivermectin was given in case of treatment failure at the end of the second week. A third dose of ivermectin, combined with permethrin 5% and salicylic acid 5% was given at the end of the fourth week for the nonresponders to the second dose. RESULTS: Two patients were completely cured after a single dose of ivermectin, 4 patients required a second dose at a 2-week interval to achieve cure and 2 patients cleared from scabies after the combined therapy. No recurrence was reported at the end of 8 weeks. An inverse relation was observed between the response to ivermectin and the severity of immunosuppression, crust thickness and mite burden. CONCLUSION: Oral ivermectin is an effective alternative therapy for the treatment of crusted scabies. The response of crusted scabies to oral ivermectin is variable and combination therapy with topical scabicides and keratolytics seems to be the best choice. PMID- 19309429 TI - Incidence of adverse cutaneous drug reactions in a Mexican sample: an exploratory study on their association to tumour necrosis factor alpha TNF2 allele. AB - BACKGROUND: Most adverse cutaneous drug reactions (ACDR) are mediated by delayed hypersensitivity (dh) with lymphocyte recruitment and inflammatory cytokines release, including tumour necrosis factor alpha (TNFalpha). Polymorphisms in the TNFalpha gene, such as the infrequent allele TNF2, predispose to certain inflammatory entities and enhance TNFalpha production. The incidence of the TNF2 allele is increased in British patients with severe ACDR, suggesting TNFalpha as a major contributor in the pathogenesis of ACDR. OBJECTIVE: We designed a prospective study to analyse the epidemiology of ACDR in a third-level Mexican hospital and explore the possibility of a relationship between the TNF2 allele and ACDR-dh. METHODS: A prospective study during 9 months allowed recognition of 34 ACDR-dh patients. The study included 33 paired patients, and 44 healthy volunteers. All subjects were genotyped for TNF2 by PCR DNA amplification and NcoI restriction endonuclease digestion. Results Incidence of ACDR was 0.95%. The TNF2 allele was detected in 9.9% of the sample population with no significant differences between healthy controls, and patients with or without ACDR-dh. Only 3 of the 34 ACDR-dh subjects presented severe reactions, with 1 having the TNF2 allele. Comorbidity analysis showed significance only with autoimmune thyroid disease, consistent with reports on Chinese and Tunisian patients. CONCLUSION: ACDR incidence and TNF2/TNFA heterozygosity were lower in Mexican than in Caucasian patients. ACDR-dh patients showed no increased frequency in the TNF2 allele. PMID- 19309430 TI - Thyroid autoimmunity in Iranian patients with pemphigus vulgaris. PMID- 19309432 TI - Seborrhoeic keratoses in patients with internal malignancies: a case-control study with prospective accrual of patients. AB - BACKGROUND: The association between the eruption of numerous seborrhoeic keratoses as a result of an underlying malignancy is controversially discussed. The aim of this case-control study with prospective accrual of patients was to determine whether a direct association exists between the number seborrhoeic keratoses and internal malignancies. METHODS: The numbers and sites of seborrhoeic keratoses were counted in 150 oncological patients and 150 matched controls. Additionally, the presence or absence of pruritus, acanthosis nigricans, and the sudden appearance of seborrhoeic keratoses were assessed. RESULTS: Seborrhoeic keratoses did not differ significantly between patients with internal malignancies and controls. Only two patients fulfilled the criteria of the Leser-Trelat sign, defined as the eruption of numerous seborrhoeic keratoses as a cutaneous marker of an underlying internal malignancy. CONCLUSION: No association was found between seborrhoeic keratoses and cancer. Furthermore, our data did not provide support to the validity of the Leser-Trelat sign in patients with internal malignancies. PMID- 19309433 TI - Prevalence and clinical features of psoriatic arthritis and joint complaints in 2009 patients with psoriasis: results of a German national survey. AB - BACKGROUND: Psoriatic arthritis (PsA) is a chronic, systemic, inflammatory disorder characterized by the association of arthritis with psoriasis. Patients with PsA may have a heterogeneous and variable clinical course. Evidence that affected patients can have significant radiographic joint damage, functional impairment, reduced quality of life and long-term work disability is increasing. OBJECTIVES: This study aims to determine the prevalence and clinical features of psoriatic arthritis and joint complaints in patients with psoriasis examined in a German national survey. METHODS: This study is a non-interventional, cross sectional analysis on 2009 patients with psoriasis from 13 dermatological hospitals and 129 dermatological private practices and outpatient clinics in Germany. Patients showing rheumatological symptoms were further recorded with respect to active arthritis and PsA symptoms according to Group for Research and Assessment of Psoriasis and Psoriatic Arthritis criteria. RESULTS: Nineteen per cent of the patients had PsA, including 14.8% previously confirmed and 4.2% newly diagnosed disease. Another 7.7% had intermittent but clinically unspecific joint symptoms, which could not be clearly attributed to PsA. About half (49.7%) of the patients with PsA had at least 1 swollen joint and 84.9% (n = 287) suffered from joint pain. Patients suffering from pain marked an average of 8.7 joints on a diagram as painful out of a possible 28. The mean number of swollen joints among the affected patients amounted to an average of 6.8. CONCLUSION: Our results show that there is still a significant number of patients suspected of having joint involvement without ever having been diagnosed with PsA. Recently published data indicate that progression of joint damage and functional disability can be prevented if adequate treatment is started promptly. Early diagnosis and interdisciplinary care are thus crucial. PMID- 19309434 TI - Versican is localized to nail mesenchyme containing onychofibroblasts. PMID- 19309435 TI - Rheumatoid leptomeningitis: magnetic resonance imaging and pathologic findings--a case report. AB - BACKGROUND AND PURPOSE: Rheumatoid arthritis (RA) is a chronic inflammatory multisystem disease with articular and extra-articular manifestations. Intracranial manifestations of RA are rare. Purpose of this article is to report on a rarely described leptomeningeal involvement in RA, and on its neuroimaging features, including diffusion-weighted imaging (DWI). METHODS: The authors describe the case of a 74-year-old woman with a 5-year history of RA presenting with progressive left-side weakness and hypoesthesia. The patient underwent laboratory investigation and brain contrast-enhanced MRI, also with DWI, before undergoing brain biopsy. RESULTS: Neuroimaging revealed abnormal high T2-signal in right frontal and parietal lobes, restricted diffusion in the subarachnoid space, and diffuse thick linear leptomeningeal contrast-enhancement. These findings were interpreted as rheumatoid leptomeningitis, and brain biopsy confirmed this diagnosis. CONCLUSIONS: In summary, rheumatoid meningitis is a rare neurological complication of RA, but it should be considered in the proper clinical setting when patient presentation and laboratory results fail to support the other differential diagnostic possibilities proposed by the MR imaging findings. PMID- 19309436 TI - Regulation of catecholamine release and tyrosine hydroxylase in human adrenal chromaffin cells by interleukin-1beta: role of neuropeptide Y and nitric oxide. AB - Adrenal chromaffin cells synthesize and secrete catecholamines and neuropeptides that may regulate hormonal and paracrine signaling in stress and also during inflammation. The aim of our work was to study the role of the cytokine interleukin-1beta (IL-1beta) on catecholamine release and synthesis from primary cell cultures of human adrenal chromaffin cells. The effect of IL-1beta on neuropeptide Y (NPY) release and the intracellular pathways involved in catecholamine release evoked by IL-1beta and NPY were also investigated. We observed that IL-1beta increases the release of NPY, norepinephrine (NE), and epinephrine (EP) from human chromaffin cells. Moreover, the immunoneutralization of released NPY inhibits catecholamine release evoked by IL-1beta. Moreover, IL 1beta regulates catecholamine synthesis as the inhibition of tyrosine hydroxylase decreases IL-1beta-evoked catecholamine release and the cytokine induces tyrosine hydroxylase Ser40 phosphorylation. Moreover, IL-1beta induces catecholamine release by a mitogen-activated protein kinase (MAPK)-dependent mechanism, and by nitric oxide synthase activation. Furthermore, MAPK, protein kinase C (PKC), protein kinase A (PKA), and nitric oxide (NO) production are involved in catecholamine release evoked by NPY. Using human chromaffin cells, our data suggest that IL-1beta, NPY, and nitric oxide (NO) may contribute to a regulatory loop between the immune and the adrenal systems, and this is relevant in pathological conditions such as infection, trauma, stress, or in hypertension. PMID- 19309437 TI - beta-N-methylamino-L-alanine induced in vivo retinal cell death. AB - Controversial debates still remain around the nature of the etiologic agent responsible for Amyotrophic lateral sclerosis/Parkinson dementia complex (ALS/PDC) whose incidence is unusually high among the population of the pacific island of Guam. It has been hypothesized that the neurotoxin beta-N-methylamino-L alanine (L-BMAA) produced by cyanobacteria in the roots of Cycas Circinalis seeds might trigger ALS/PDC. Frequently observed in patients with ALS/PDC, retinopathy is one of the clinical features of the disease. The effect of the L-BMAA on cell viability was examined in vivo by measuring the electrophysiological activity of the mouse retinal neurons by electroretinography recordings. Intra-ocular injections of L-BMAA selectively reduced the b-wave amplitude, without affecting neither the a-wave amplitude nor the a- and b-latencies. The cell death of retinal cells was evidenced by histology on retina sections, caspase 3 activation, incorporation of propidium iodide and production of reactive oxygen species. Co-injection with the specific NMDA antagonist, MK-801, significantly protected the retinal neurons from L-BMAA/NMDA-induced apoptosis. We provide evidence that L-BMAA induced neuronal cell death in vivo supporting a direct causal link between L-BMAA and neuronal damages. PMID- 19309438 TI - Enhanced ovicidal activity of an oil formulation of the fungus Metarhizium anisopliae on the mosquito Aedes aegypti. AB - The effect of humidity on the activity of Metarhizium anisopliae IP 46 (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) formulated in sunflower oil against Aedes aegypti (L.) (Diptera: Culicidae) eggs was examined. After exposure of eggs at 75% relative humidity (RH) for 98% RH, eclosion was 98% RH, followed by: (a) a 12-day exposure at 75% RH before submersion in water; (b) a minimal 5-day exposure at > 98% RH and direct subsequent transfer of treated eggs to water, or (c) a minimal daily 20-h exposure at > 98% RH alternating with 4 h at 75% RH for 10 days. We demonstrate that oil-based formulations of conidia of M. anisopliae enhance ovicidal activity at high humidities and conclude that these formulations have potential in the integrated control of Ae. aegypti. PMID- 19309439 TI - Age-dependent association of idiopathic achalasia with vasoactive intestinal peptide receptor 1 gene. AB - Idiopathic achalasia is a rare disorder of the oesophagus of unknown aetio pathogenesis characterized by a myenteric inflammation, aperistalsis and insufficient lower oesophageal sphincter relaxation. Vasoactive intestinal peptide (VIP), present in the myenteric plexus, is involved in smooth muscle relaxation and acts as an anti-inflammatory cytokine. The human VIP receptor 1 gene (VIPR1) is highly polymorphic and may play a role in idiopathic achalasia. One hundred and four consecutive patients and 300 random controls from the same geographic area were typed for five SNPs mapping in the VIPR1 gene. Patients with idiopathic achalasia show a significant difference in allele, genotype and phenotype distribution of SNP rs437876 mapping in intron 4. This association, however, was almost entirely due to the group of patients with late disease onset (P = 0.0005). These results strongly suggest that idiopathic achalasia is a heterogeneous disease with a different aetiology in cases with early or late disease onset. PMID- 19309440 TI - Optimizing analysis of stable isotope breath tests to estimate gastric emptying of solids. AB - Breath tests (BT) using 13C-substrates have been proposed for the measurement of gastric emptying (GE). The mathematical analysis of the breath 13CO2 excretion that most accurately predicts GE t(1/2) from simultaneous scintigraphy is unresolved. To compare five mathematical methods to estimate GE t(1/2) by BT with t(1/2) from simultaneous scintigraphy. Data acquired from a dual-labelled solid liquid meal containing 99mTc sulphur colloid and 13C-Spirulina platensis from 57 healthy volunteers were used to compare four mathematical methods reported in the literature [Ghoos method; generalized linear regression (Viramontes); linear regression (Szarka); Wagner-Nelson method] and the total cumulative breath 13CO2 excretion with > or =12 breath samples collected over at least 4 h. The concordance correlation coefficient (CCC) for the t(1/2) results obtained with each method using BT data was compared with the results obtained with scintigraphy. The linear regression and generalized linear regression methods used five samples at 45, 90, 120, 150 and 180 min. All methods, except for the Wagner-Nelson method, resulted in mean GE t(1/2) that approximated t(1/2) obtained with scintigraphy. The highest CCC was observed with the linear regression method. Simple cumulative excretion of breath 13CO2 provides a better CCC than the Ghoos method. The linear regression and generalized linear regression methods (which also require relatively few breath samples) provide the most accurate analyses of breath 13CO2 excretion in stable isotope GEBT. PMID- 19309441 TI - Lactobacillus farciminis treatment attenuates stress-induced overexpression of Fos protein in spinal and supraspinal sites after colorectal distension in rats. AB - Abstract Irritable bowel syndrome (IBS), frequently associated with psychological distress, is characterized by hypersensitivity to gut wall distension. Some probiotics are able to alleviate IBS symptoms and reduce visceromotor response to mechanical stimuli in animals. Moreover, we have previously shown that Lactobacillus farciminis treatment abolished the hyperalgesia to colorectal distension (CRD) induced by acute stress. The aims of the present study were to determine whether (i) stress-induced visceral hyperalgesia modifies the expression of Fos, a marker of general neuronal activation, induced by CRD, (ii) this activation can be modulated by L. farciminis treatment. Female rats were treated by L. farciminis and CRD was performed after partial restraint stress (PRS) or sham-PRS. The expression of Fos protein was measured by immunohistochemistry. After CRD or PRS, Fos expression was increased in spinal cord section (S1), nucleus tractus solitarius (NTS), paraventricular nucleus (PVN) of the hypothalamus, and in the medial nucleus of the amygdala (MeA). The combination of both stimuli, PRS and CRD, markedly increased this Fos overexpression in the sacral spinal cord section, PVN and MeA, but not in NTS. By contrast, a pretreatment with L. farciminis significantly reduced the number of Fos positive cells in these area. This study shows that PRS enhances Fos protein expression induced by CRD at the spinal and supraspinal levels in rats. Lactobacillus farciminis treatment inhibited this enhancing effect, suggesting that the antinociceptive effect of this probiotic strain results from a decrease of the stress-induced activation/sensitization of sensory neurons at the spinal and supraspinal level. PMID- 19309442 TI - 5-hydroxyindalpine, an agonist at the putative 5-HT receptor, has no activity on human recombinant monoamine receptors but accelerates distension-induced peristalsis in mouse isolated colon. AB - Although the putative 5-HT(1P) receptor has been implicated to have a role in peristalsis, experiments which suggest this function are preliminary or have measured only components of the reflex. We have, therefore, further characterized a reported agonist at this receptor (5-hydroxyindalpine; 5-OHIP) and investigated the effects of 5-OHIP and 5-hydroxytrytophan-dipeptide (5-HTP-DP), a reported 5 HT(1P) receptor antagonist, on distension-induced peristalsis in mouse colon. The effects of 5-OHIP on intracellular calcium, cyclic adenosine monophosphate concentrations or GTPgammaS binding were measured in cell lines expressing human recombinant 5-HT(1A, 1B, 1D, 2A, 2B, 2C, 3, 4, 6, 7) and alpha(1A), alpha(1B), D(1), D(2), D(3), H(1), H(3) receptors. The effects of 5-OHIP and 5-HTP-DP on peristalsis were assessed by measuring changes in frequency and times to reach threshold of peristaltic contractions, as well as the threshold and maximum pressures of each peristaltic stroke. 5-hydroxyindalpine (1 nmol L(-1)-10 micromol L(-1)) had no significant activity at any of the receptors studied. However, 5-OHIP (0.1 nmol L(-1)-1 micromol L(-1)) concentration-dependently increased the frequency of peristalsis (EC(50) = 4.4 nmol L(-1)) and reduced the time taken to reach threshold and threshold pressure, without altering maximum pressures. The maximum effect of 5-OHIP was at 1 micromol L(-1) (68.0 +/- 14.5% increase in frequency); 10 micromol L(-1) decreased peristalsis. 5 hydroxytrytophan-dipeptide (1-300 nmol L(-1)) also increased the frequency of peristalsis and prevented the actions of 5-OHIP. The higher concentration (1 micromol L(-1)) transiently inhibited peristalsis and after recovery, prevented the actions of 5-OHIP but not the excitatory activity of the cholinesterase inhibitor neostigmine. In summary, the present data demonstrate an interaction of '5-HT(1P)-ligands' with the peristaltic reflex. However, the absence of an effect of 5-OHIP on a range of different monoamine receptors continues to highlight the need to investigate the identity of the putative 5-HT(1P) receptor. PMID- 19309443 TI - Coupling and propagation of normal and dysrhythmic gastric slow waves during acute hyperglycaemia in healthy humans. AB - Acute hyperglycaemia in healthy humans disrupts slow wave rhythm similar to that observed in diabetic gastropathy, but hyperglycaemic effects on regional dysrhythmias, power, coupling and propagation are unexplored. Using mucosal mapping, we aimed to demonstrate that hyperglycaemia elicits region-specific tachygastria and evokes slow wave uncoupling between adjacent regions. Catheters with bipolar electrodes were affixed 10.5, 6 and 2 cm from the pylorus during endoscopy with midazolam in 10 healthy humans. Recordings were obtained for 1 h under basal conditions and for 1 h with hyperglycaemic clamping to 250 mg dL(-1). In basal recordings, proximal and distal slow wave frequencies were similar [2.91 +/- 0.05 vs 2.81 +/- 0.09 cycles per minute (cpm)]. Tachygastria (>3.6 cpm) was present 1.7 +/- 1.1% of the time proximally and 3.3 +/- 1.8% distally and localized to one lead 67% of the time. Proximal to distal gradients in signal power and power variability were observed. Coupling between adjacent sites was 78 +/- 2% with propagation velocities of 1.3 +/- 0.1 cm s(-1). 2 +/- 1% of segments showed >50% uncoupling. Hyperglycaemic clamping increased mean proximal (3.18 +/- 0.11 cpm) and distal (3.50 +/- 0.12 cpm) frequencies and proximal (15 +/- 6%) and distal (32 +/- 9%) tachygastria (all P < 0.01) that localized to one lead 80% of the time. During periods of normal frequency, coupling decreased proximally (54 +/- 6%) and distally (47 +/- 4%) (P < 0.01). 55 +/- 8% of segments showed >50% uncoupling (P < 0.01). In conclusion, gastric slow waves show stable, highly coupled rhythms under basal conditions. Hyperglycaemia elicits isolated tachygastrias and uncoupling of normal slow waves that are most prominent distally. These findings provide a foundation for studying slow wave conduction defects in diabetic gastropathy. PMID- 19309445 TI - A major quantitative trait locus controlling cadmium translocation in rice (Oryza sativa). AB - The trait of low cadmium (Cd) accumulation in brown rice (Oryza sativa) is important for food safety. An effective way to reduce Cd accumulation in the grain is to control Cd transfer from the roots to the shoots. Here, we investigated genotypic variation in the shoot Cd concentration among 146 accessions from a rice core collection and performed a quantitative trait locus (QTL) analysis to determine the loci controlling shoot Cd accumulation. Furthermore, we physiologically characterized the two accessions used for QTL analysis. Large genotypic variation (13-fold) in the shoot Cd concentration was found. A major QTL was detected on chromosome 11 using a F2 population derived from Badari Dhan (a high-Cd accession) and Shwe War (a low-Cd accession). This QTL explained 16.1% of the phenotypic variation in Cd accumulation. Furthermore, this QTL was confirmed by analysis of advanced progeny. Physiological studies showed that Badari Dhan and Shwe War did not differ in uptake of Cd by the roots, but differed greatly in the translocation of Cd from the roots to the shoots. Taken together, our findings suggest that the major QTL detected is responsible for the translocation of Cd from the roots to the shoots. PMID- 19309446 TI - Isotopic evidence of full and partial myco-heterotrophy in the plant tribe Pyroleae (Ericaceae). AB - Botanists and mycologists have long debated the potential for full myco heterotrophy in the achlorophyllous Pyrola aphylla (Ericaceae). Here we address the ecophysiology of this putative myco-heterotroph and two other closely related green species in the tribe Pyroleae (Pyrola picta and Chimaphila umbellata). The stable isotopes of carbon and nitrogen (delta13C and delta15N) were analysed from 10 populations of Pyroleae species in California and Oregon, USA. For all populations isotope signatures were tested for significant differences between P. aphylla, green pyroloids, surrounding autotrophs and obligate myco-heterotrophs. Throughout all populations P. aphylla was most similar to myco-heterotrophs that associate with ectomycorrhizal fungi in its 13C signature (average enrichment epsilon13C = 6.9 +/- 0.9 per thousand) and even more enriched in 15N than many previously recorded myco-heterotrophic species (average enrichment epsilon15N = 18.0 +/- 2.2 per thousand). The two green Pyroleae species were not enriched in 13C compared with the autotrophic understory (C. umbellata average enrichment epsilon13C = -0.5 +/- 1.0 per thousand and P. picta average epsilon13C = 0.3 +/- 1.4 per thousand) and their 15N signatures were similar to myco-heterotrophs that associate with ectomycorrhizal fungi (C. umbellata average enrichment epsilon15N = 10.6 +/- 1.6 per thousand and P. picta average epsilon15N = 10.6 +/- 1.9 per thousand). This is the first study to analyse the isotope signatures of P. aphylla from a wide geographic region and our results confirm the variable trophic strategies of adult plants within the Pyroleae and the myco-heterotrophic status of P. aphylla. PMID- 19309447 TI - Embolism resistance of three boreal conifer species varies with pit structure. AB - While tracheid size of conifers is often a good proxy of water transport efficiency, correlations between conifer wood structure and transport safety remain poorly understood. It is hypothesized that at least some of the variation in bordered pit and tracheid structure is associated with both transport efficiency and embolism resistance. Stem and root samples from three boreal Pinaceae species were collected to test this hypothesis. Tracheid and pit anatomy were studied using light microscopy as well as scanning and transmission electron microscopy. While tracheid size explained at least 90% of the variation in specific conductivity for stem and root samples, the strongest correlations with embolism resistance occurred at the pit level. Both torus thickness and depth of the pit chamber showed a linear increase with greater vulnerability to cavitation. Greater embolism resistance was correlated with increasing wood density and tracheid wall reinforcement. A thinner torus may be more flexible and better able to seal the pit aperture. The pit chamber depth is proportional to the distance that the margo needs to deflect for pit aspiration. PMID- 19309448 TI - Taxon recruitment of the arctic flora: an analysis of phylogenies. AB - The Arctic is the endpoint of many climatic gradients and is presently occupied by c. 2200 vascular plant species. Glaciation started in the Middle Eocene but a significant expansion of the Greenland ice shield occurred only c. 3.2 million yr ago, leading to the expansion of the treeless circumpolar arctic tundra. Available molecular phylogenetic studies were evaluated for 148 of 374 genera occurring in the Arctic to determine the relative roles of their independent origins and their diversification in the development of the contemporary arctic flora. The number of arctic species paralleled the total number of species in a genus. Multiple arctic species within a genus originated mostly independently of each other and from different lineages. Minor radiations occurred in only a few genera and major radiations were absent. Mostly parallel evolution of arctic taxa from nonarctic ancestors, supposedly of different ages of origin, scarcity of radiations and rarity of endemics are main features of the arctic flora. PMID- 19309449 TI - Mild fasting hyperglycemia in children: high rate of glucokinase mutations and some risk of developing type 1 diabetes mellitus. AB - BACKGROUND: Incidental hyperglycemia in children generates concern about the presence of preclinical type 1 diabetes mellitus (T1DM). OBJECTIVE: To genetically evaluate two common forms of maturity-onset diabetes of youth (MODY), the short-term prognosis in children with mild hyperglycemia, and a positive family history of diabetes mellitus. SUBJECTS: Asymptomatic children and adolescents (n = 14), younger than 15 yr, with fasting hyperglycemia, a positive family history of mild non-progressive hyperglycemia, and negative pancreatic autoantibodies were studied. PATIENTS AND METHODS: Glucokinase gene (GCK) and hepatocyte nuclear factor 1 alpha gene (HNF1A) causing two common forms of MODY were sequenced. The clinical outcome was evaluated after a follow-up period of 2.8 +/- 1.3 yr. RESULTS: GCK mutations were present in seven children. The confirmation of this diagnosis allowed discontinuation of insulin in two families and oral medications in three families. Mutations of HNF1A were not detected in any of the families. During the follow-up period, all the GCK mutation carrier children remained asymptomatic without medication and the last hemoglobin A1c levels were 6.4 +/- 0.7%. In the GCK-negative children (n = 7), one developed T1DM, corresponding to 7.2% of the total group. Mild fasting hyperglycemia persisted during follow-up in four GCK-negative children and normalized in the remaining two. CONCLUSIONS: The presence of mild persistent hyperglycemia in any patient without autoantibodies should lead to genetic analysis of GCK, particularly if there is a positive family history. Furthermore, those without GCK mutations should be followed with repeat autoantibody testing, and other genetic types of diabetes should be considered if hyperglycemia worsens. PMID- 19309450 TI - Practical matters, rather than lack of trust, motivate non-participation in a long-term cohort trial. AB - OBJECTIVE: The objective of this study was to investigate the importance of trust in researchers and other reasons that participating parents, former participants, and non-participants had for participating, or not participating, in a longitudinal cohort study on prediction and development of diabetes in children. STUDY DESIGN: A questionnaire addressing each of these groups, where respondents graded the importance of a set of listed reasons for participating/not participating, was randomly distributed to 2500 families in the All Babies in Southeast Sweden (ABIS) study region with children born between 1997 and 1999. RESULTS: Lack of trust was not a central factor to a great majority of respondents who decided not to participate in the ABIS study or who later decided to opt out. Practical matters, like blood sampling and lack of time, were important factors to many more. Yet, four fifths of those who still participate in the ABIS study stated trust in the researchers to be an important factor to their initial decision to participate. CONCLUSIONS: Trust in researchers may be a necessary prerequisite in order for people to be willing to participate in research, but practical matters such as time that has to be spent or pain involved in collecting blood were more important factors than lack of trust in explaining opt out in relation to the ABIS study. PMID- 19309452 TI - Long-term outcome of intensive initial immunosuppression protocol in pediatric deceased donor renal transplantation. AB - To report the long-term outcome of deceased donor kidney transplantation in children with emphasis on the use of an intensive initial immunosuppression protocol using R-ATG as antibody induction. Between January 1991 and December 1997, 82 deceased donor kidney transplantations were performed in 75 pediatric recipients. Mean recipient age at transplantation was 12.9 yr and the mean follow up period was 12.6 yr. All patients received quadruple immunosuppression with steroid, cyclosporine, azathioprine, and antibody induction using R-ATG Fresenius. Actual one, five, and 10 yr patient survival rates were 99%, 97%, and 94%, respectively; only one patient (1.2%) developed PTLD. Actual one, five, and 10 yr overall graft survival rates were 84%, 71%, and 50%, respectively; there were five cases (6%) of graft thrombosis and the actual immunological graft survival rates were 91%, 78%, and 63% at one, five, and 10 yr, respectively. The use of an intensive initial immunosuppression protocol with R-ATG as antibody induction is safe and effective in pediatric recipients of deceased donor kidneys with excellent immunological graft survival without an increase in PTLD or other neoplasms over a minimum 10-yr follow up. PMID- 19309453 TI - Auxin-induced, SCF(TIR1)-mediated poly-ubiquitination marks AUX/IAA proteins for degradation. AB - The plant hormone auxin (indole-3-acetic acid or IAA) regulates plant development by inducing rapid cellular responses and changes in gene expression. Auxin promotes the degradation of Aux/IAA transcriptional repressors, thereby allowing auxin response factors (ARFs) to activate the transcription of auxin-responsive genes. Auxin enhances the binding of Aux/IAA proteins to the receptor TIR1, which is an F-box protein that is part of the E3 ubiquitin ligase complex SCF(TIR1). Binding of Aux/IAA proteins leads to degradation via the 26S proteasome, but evidence for SCF(TIR1)-mediated poly-ubiquitination of Aux/IAA proteins is lacking. Here we used an Arabidopsis cell suspension-based protoplast system to find evidence for SCF(TIR1)-mediated ubiquitination of the Aux/IAA proteins SHY2/IAA3 and BDL/IAA12. Each of these proteins showed a distinct abundance and repressor activity when expressed in this cell system. Moreover, the amount of endogenous TIR1 protein appeared to be rate-limiting for a proper auxin response measured by the co-transfected DR5::GUS reporter construct. Co-transfection with 35S::TIR1 led to auxin-dependent degradation, and excess of 35S::TIR1 even led to degradation of Aux/IAAs in the absence of auxin treatment. Expression of the mutant tir1-1 protein or the related F-box protein COI1, which is involved in jasmonate signaling, had no effect on Aux/IAA degradation. Our results show that SHY2/IAA3 and BDL/IAA12 are poly-ubiquitinated and degraded in response to increased auxin or TIR1 levels. In conclusion, our data provide experimental support for the model that SCF(TIR1)-dependent poly-ubiquitination of Aux/IAA proteins marks these proteins for degradation by the 26S proteasome, leading to activation of auxin-responsive gene expression. PMID- 19309454 TI - Polyadenylation in Arabidopsis and Chlamydomonas organelles: the input of nucleotidyltransferases, poly(A) polymerases and polynucleotide phosphorylase. AB - The polyadenylation-stimulated RNA degradation pathway takes place in plant and algal organelles, yet the identities of the enzymes that catalyze the addition of the tails remain to be clarified. In a search for the enzymes responsible for adding poly(A) tails in Chlamydomonas and Arabidopsis organelles, reverse genetic and biochemical approaches were employed. The involvement of candidate enzymes including members of the nucleotidyltransferase (Ntr) family and polynucleotide phosphorylase (PNPase) was examined. For several of the analyzed nuclear-encoded proteins, mitochondrial localization was established and possible dual targeting to mitochondria and chloroplasts could be predicted. We found that certain members of the Ntr family, when expressed in bacteria, displayed poly(A) polymerase (PAP) activity and partially complemented an Escherichia coli strain lacking the endogenous PAP1 enzyme. Other Ntr proteins appeared to be specific for tRNA maturation. When the expression of PNPase was down-regulated by RNAi in Chlamydomonas, very few poly(A) tails were detected in chloroplasts for the atpB transcript, suggesting that this enzyme may be solely responsible for chloroplast polyadenylation activity in this species. Depletion of PNPase did not affect the number or sequence of mitochondrial mRNA poly(A) tails, where unexpectedly we found, in addition to polyadenylation, poly(U)-rich tails. Together, our results identify several Ntr-PAPs and PNPase in organelle polyadenylation, and reveal novel poly(U)-rich sequences in Chlamydomonas mitochondria. PMID- 19309455 TI - The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins. AB - Discovery of the jasmonate ZIM-domain (JAZ) repressors defined the core jasmonate (JA) signalling module as COI1-JAZ-MYC2, and allowed a full view of the JA signalling pathway from hormone perception to transcriptional reprogramming. JAZ proteins are repressors of MYC2 and targets of SCF(COI1), which is the likely jasmonate receptor. Upon hormone perception, JAZ repressors are degraded by the proteasome releasing MYC2 and allowing the activation of JA responses. All members of the JAZ family share two conserved domains, the Jas motif, required for JAZ interactions with MYC2 and COI1, and the ZIM domain, the function of which is so far unknown. Here, we show that the ZIM domain acts as a protein protein interaction domain mediating homo- and heteromeric interactions between JAZ proteins. These JAZ-JAZ interactions are independent of the presence of the hormone. The observation that only a few members of the JAZ family form homo- and heteromers may suggest the relevance of these proteins in the regulation of JA signalling. Interestingly, the JAZ3DeltaJas protein interacts with several JAZ proteins, providing new clues to understanding the dominant JA insensitivity promoted by truncated JAZDeltaJas proteins. We also provide evidence that the Jas motif mediates the hormone-dependent interaction between Arabidopsis JAZ3 and COI1, and further confirm that the Jas motif is required and sufficient for Arabidopsis JAZ3-MYC2 interaction. Finally, we show that interaction with MYC2 is a common feature of the JAZ family, as most JAZ proteins can bind MYC2 in pull down and yeast two-hybrid assays. PMID- 19309456 TI - Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. AB - Plant membrane compartments and trafficking pathways are highly complex, and are often distinct from those of animals and fungi. Progress has been made in defining trafficking in plants using transient expression systems. However, many processes require a precise understanding of plant membrane trafficking in a developmental context, and in diverse, specialized cell types. These include defense responses to pathogens, regulation of transporter accumulation in plant nutrition or polar auxin transport in development. In all of these cases a central role is played by the endosomal membrane system, which, however, is the most divergent and ill-defined aspect of plant cell compartmentation. We have designed a new vector series, and have generated a large number of stably transformed plants expressing membrane protein fusions to spectrally distinct, fluorescent tags. We selected lines with distinct subcellular localization patterns, and stable, non-toxic expression. We demonstrate the power of this multicolor 'Wave' marker set for rapid, combinatorial analysis of plant cell membrane compartments, both in live-imaging and immunoelectron microscopy. Among other findings, our systematic co-localization analysis revealed that a class of plant Rab1-homologs has a much more extended localization than was previously assumed, and also localizes to trans-Golgi/endosomal compartments. Constructs that can be transformed into any genetic background or species, as well as seeds from transgenic Arabidopsis plants, will be freely available, and will promote rapid progress in diverse areas of plant cell biology. PMID- 19309457 TI - Transient expression in Nicotiana benthamiana fluorescent marker lines provides enhanced definition of protein localization, movement and interactions in planta. AB - Here, we report on the construction of a novel series of Gateway-compatible plant transformation vectors containing genes encoding autofluorescent proteins, including Cerulean, Dendra2, DRONPA, TagRFP and Venus, for the expression of protein fusions in plant cells. To assist users in the selection of vectors, we have determined the relative in planta photostability and brightness of nine autofluorescent proteins (AFPs), and have compared the use of DRONPA and Dendra2 in photoactivation and photoconversion experiments. Additionally, we have generated transgenic Nicotiana benthamiana lines that express fluorescent protein markers targeted to nuclei, endoplasmic reticulum or actin filaments. We show that conducting bimolecular fluorescence complementation assays in plants that constitutively express cyan fluorescent protein fused to histone 2B provides enhanced data quality and content over assays conducted without the benefit of a subcellular marker. In addition to testing protein interactions, we demonstrate that our transgenic lines that express red fluorescent protein markers offer exceptional support in experiments aimed at defining nuclear or endomembrane localization. Taken together, the new combination of pSITE-BiFC and pSITEII vectors for studying intracellular protein interaction, localization and movement, in conjunction with our transgenic marker lines, constitute powerful tools for the plant biology community. PMID- 19309458 TI - Functional expression and characterization of Arabidopsis ABCB, AUX 1 and PIN auxin transporters in Schizosaccharomyces pombe. AB - Heterologous expression systems based on tobacco BY-2 cells, Arabidopsis cell cultures, Xenopus oocytes, Saccharomyces cerevisiae, and human HeLa cells have been used to express and characterize PIN, ABCB (PGP), and AUX/LAX auxin transporters from Arabidopsis. However, no single system has been identified that can be used for effective comparative analyses of these proteins. We have developed an accessible Schizosaccharomyces pombe system for comparative studies of plant transport proteins. The system includes knockout mutants in all ABC and putative auxin transport genes and Gateway((R))-compatible expression vectors for functional analysis and subcellular localization of recombinant proteins. We expressed Arabidopsis ABCB1 and ABCB19 in mam1pdr1 host lines under the inducible nmt41 promoter. ABCB19 showed a higher (3)H-IAA export activity than ABCB1. Arabidopsis PIN proteins were expressed in a mutant lacking the auxin effluxer like 1 (AEL1) gene. PIN1 showed higher activity than PIN2 with similar protein expression levels. Expression of AUX1 in a permease-deficient vat3 mutant resulted in increased net auxin uptake activity. Finally, ABCB4 expressed in mam1pdr1 displayed a concentration-dependent reversal of (3)H-IAA transport that is consistent with its observed activity in planta. Structural modelling suggests that ABCB4 has three substrate interaction sites rather than the two found in ABCB19, thus providing a rationale for the observed substrate activation. Taken together, these results suggest that the S. pombe system described here can be employed for comparative analyses and subsequent structural characterizations of plant transport proteins. PMID- 19309459 TI - Characterization of selenocysteine methyltransferases from Astragalus species with contrasting selenium accumulation capacity. AB - A group of selenium (Se)-hyperaccumulating species belonging to the genus Astragalus are known for their capacity to accumulate up to 0.6% of their foliar dry weight as Se, with most of this Se being in the form of Se methylselenocysteine (MeSeCys). Here, we report the isolation and molecular characterization of the gene that encodes a putative selenocysteine methyltransferase (SMT) enzyme from the non-accumulator Astragalus drummondii and biochemically compare it with an authentic SMT enzyme from the Se hyperaccumulator Astragalus bisulcatus, a related species that lives within the same native habitat. The non-accumulator enzyme (AdSMT) shows a high degree of homology with the accumulator enzyme (AbSMT) but lacks the selenocysteine methyltransferase activity in vitro, explaining why little or no detectable levels of MeSeCys accumulation are observed in the non-accumulator plant. The insertion of mutations on the coding region of the non-accumulator AdSMT enzyme to better resemble enzymes that originate from Se accumulator species results in increased selenocysteine methyltransferase activity, but these mutations were not sufficient to fully gain the activity observed in the AbSMT accumulator enzyme. We demonstrate that SMT is localized predominantly within the chloroplast in Astragalus, the principal site of Se assimilation in plants. By using a site directed mutagenesis approach, we show that an Ala to Thr amino acid mutation at the predicted active site of AbSMT results in a new enzymatic capacity to methylate homocysteine. The mutated AbSMT enzyme exhibited a sixfold higher capacity to methylate selenocysteine, thereby establishing the evolutionary relationship of SMT and homocysteine methyltransferase enzymes in plants. PMID- 19309460 TI - Identification of the Arabidopsis dry2/sqe1-5 mutant reveals a central role for sterols in drought tolerance and regulation of reactive oxygen species. AB - Squalene epoxidase enzymes catalyse the conversion of squalene into 2,3 oxidosqualene, the precursor of cyclic triterpenoids. Here we report that the Arabidopsis drought hypersensitive/squalene epoxidase 1-5 (dry2/sqe1-5) mutant, identified by its extreme hypersensitivity to drought stress, has altered stomatal responses and root defects because of a point mutation in the SQUALENE EPOXIDASE 1 (SQE1) gene. GC-MS analysis indicated that the dry2/sqe1-5 mutant has altered sterol composition in roots but wild-type sterol composition in shoots, indicating an essential role for SQE1 in root sterol biosynthesis. Importantly, the stomatal and root defects of the dry2/sqe1-5 mutant are associated with altered production of reactive oxygen species. As RHD2 NADPH oxidase is de localized in dry2/sqe1-5 root hairs, we propose that sterols play an essential role in the localization of NADPH oxidases required for regulation of reactive oxygen species, stomatal responses and drought tolerance. PMID- 19309461 TI - Asymmetric allele-specific expression in relation to developmental variation and drought stress in barley hybrids. AB - In the present study, we analysed allele-specific expression (ASE) in the selfing species barley to assess the frequency of cis-acting regulatory variation and the effects of genetic background, developmental differences and drought stress on allelic expression levels. We measured ASE ratios in 30 genes putatively involved in stress responses in five hybrids and their reciprocals, namely Hordeum spontaneum 41-1/Alexis (HAl), Hordeum spontaneum 41-1/Arta (HAr), Sloop/WI3408 (SW), Tadmor/Sloop (TS) and Tadmor/WI3408 (TW). In order to detect cis-acting variation related to drought and developmental changes, the barley hybrids were grown under control and water-limited conditions, and leaf tissue was harvested at two developmental stages. The analysis demonstrated that more than half of the genes measured (63%) showed allelic differences in expression of up to 19-fold due to cis-regulatory variation in at least one cross by treatment/stage combination. Drought stress induced changes in allelic expression ratios, indicating differences between drought responsive cis-elements. In addition, ASE differences between developmental stages suggested the presence of cis-acting elements interacting with developmental cues. We were also able to demonstrate that the levels and frequency of allelic imbalance and hence differences in cis regulatory elements are correlated with the genetic divergence between the parental lines, but may also arise as an adaptation to diverse habitats. Our findings suggest that cis-regulatory variation is a common phenomenon in barley, and may provide a molecular basis of transgression. Differential expression of near-isogenic members of the same gene family could potentially result in hybrid lines out performing their parents in terms of expression level, timing and response to developmental and environmental cues. Identification and targeted manipulation of cis-regulatory elements will assist in breeding improved crops with a better adaptation to changing environments. PMID- 19309462 TI - The MYB transcription factor GhMYB25 regulates early fibre and trichome development. AB - Little is still known about the developmental control of the long seed coat trichomes of cotton (Gossypium hirsutum L.). In Arabidopsis, leaf trichome initiation is regulated by a group of well-defined transcription factors that includes MYB and homeodomain types. Many MYBs are expressed in fibres, but their roles in fibre development remain unclear. We analysed the function of one MYB transcription factor, GhMYB25, identified from transcriptome comparisons between wild-type and fibreless cotton mutants. A GhMYB25 promoter-GUS construct in transgenic cotton was expressed in the epidermis of ovules, developing fibre initials and fibres, in the trichomes of a number of tissues including leaves, stems and petals, as well as in the anthers, pollen and the epidermal layers of roots and root initials, but not in root hairs. Cotton is an allotetraploid with two very similar GhMYB25 genes that were silenced by a single RNAi construct. GhMYB25-silenced cotton showed alterations in the timing of rapid fibre elongation, resulting in short fibres, dramatic reductions in trichomes on other parts of the plant, and reductions in seed production. Reciprocal crosses between transgenic and non-transgenic plants indicated that pollen and ovule viability per se were not disrupted. Ectopic over-expression of GhMYB25 had more subtle impacts, with increases in cotton fibre initiation and leaf trichome number. High expression appeared to adversely affect fertility. Our results provide convincing evidence for a role of GhMYB25, like other MIXTA-like MYBS, in regulating specialized outgrowths of epidermal cells, including, in this case, cotton fibres. PMID- 19309463 TI - Identification of a novel E3 ubiquitin ligase that is required for suppression of premature senescence in Arabidopsis. AB - During leaf senescence, resources are recycled by redistribution to younger leaves and reproductive organs. Candidate pathways for the regulation of onset and progression of leaf senescence include ubiquitin-dependent turnover of key proteins. Here, we identified a novel plant U-box E3 ubiquitin ligase that prevents premature senescence in Arabidopsis plants, and named it SENESCENCE ASSOCIATED E3 UBIQUITIN LIGASE 1 (SAUL1). Using in vitro ubiquitination assays, we show that SAUL1 has E3 ubiquitin ligase activity. We isolated two alleles of saul1 mutants that show premature senescence under low light conditions. The visible yellowing of leaves is accompanied by reduced chlorophyll content, decreased photochemical efficiency of photosystem II and increased expression of senescence genes. In addition, saul1 mutants exhibit enhanced abscisic acid (ABA) biosynthesis. We show that application of ABA to Arabidopsis is sufficient to trigger leaf senescence, and that this response is abolished in the ABA insensitive mutants abi1-1 and abi2-1, but enhanced in the ABA-hypersensitive mutant era1-3. We found that increased ABA levels coincide with enhanced activity of Arabidopsis aldehyde oxidase 3 (AAO3) and accumulation of AAO3 protein in saul1 mutants. Using label transfer experiments, we showed that interactions between SAUL1 and AAO3 occur. This suggests that SAUL1 participates in targeting AAO3 for ubiquitin-dependent degradation via the 26S proteasome to prevent premature senescence. PMID- 19309464 TI - Illusory control: a generative force behind power's far-reaching effects. AB - Three experiments demonstrated that the experience of power leads to an illusion of personal control. Regardless of whether power was experientially primed (Experiments 1 and 3) or manipulated through roles (manager vs. subordinate; Experiment 2), it led to perceived control over outcomes that were beyond the reach of the power holder. Furthermore, this illusory control mediated the influence of power on several self-enhancement and approach-related outcomes reported in the power literature, including optimism (Experiment 2), self-esteem (Experiment 3), and action orientation (Experiment 3). These results demonstrate the theoretical importance of perceived control as a generative cause of and driving force behind many of power's far-reaching effects. A fourth experiment ruled out an alternative explanation: that positive mood, rather than illusory control, is at the root of power's effects. The discussion considers implications for existing and future research on the psychology of power, perceived control, and positive illusions. PMID- 19309465 TI - Transformation direction influences shape-similarity judgments. AB - Three experiments provide evidence that the perceived similarity between two images is systematically affected by the inherent direction of a transformation that links the two. Participants were shown short animations morphing one object into another from the same basic category. They were then asked to make directional similarity judgments ("How similar is object A to object B?") for two stationary images drawn from the morph continuum. Across three experiments, similarity ratings for identical comparisons were higher when the reference object, B, had appeared before the comparison object, A, in the preceding morph sequence. This response to dynamic transformational sequences is in accordance with the view that similarity depends on the ease of transformation between object representations and that transformations between objects in categorization and object recognition are psychologically real. PMID- 19309466 TI - Panic search: fear produces efficient visual search for nonthreatening objects. PMID- 19309467 TI - First service pregnancy rates following post-AI use of HCG in Ovsynch and Heatsynch programmes in lactating dairy cows. AB - Lactating dairy cows (n = 667) at random stages of the oestrous cycle were assigned to either ovsynch (O, n = 228), heatsynch (H, n = 252) or control (C, n = 187) groups. Cows in O and H groups received 100 microg of GnRH agonist, i.m. (day 0) starting at 44 +/- 3 days in milk (DIM), and 500 microg of cloprostenol, i.m. (day 7). In O group, cows received 100 microg of GnRH (day 9) and were artificially inseminated without oestrus detection 16-20 h later. In H group, cows received 1 mg oestradiol benzoate (EB) i.m., 24 h after the cloprostenol injection and were artificially inseminated without oestrus detection 48-52 h after the EB injection. Cows in C group were inseminated at natural oestrus. On the day of artificial insemination (AI), cows in all groups were assigned to subgroups as follows: human Chorionic Gonadotrophin (O-hCG) (n = 112), O-saline (n = 116), H-hCG (n = 123), H-saline (n = 129), C-hCG (n = 94) and C-saline (n = 93) subgroups. Cows in hCG and saline subgroups received 3000 IU hCG i.m. and or 10 ml saline at day 5 post-AI (day 15), respectively. Pregnancy status was assessed by palpation per rectum at days 40 to 45 after AI. The logistic regression model using just main effects of season (summer and winter), parity (primiparous and pluriparous), method(1) (O, H and C) and method(2) (hCG and saline) showed that all factors, except method(1), were significant. Significant effects of season (p < 0.01), hCG and parity (p < 0.01), and a trend of parity and season (p < 0.1) were detected. A clear negative effect of warm period on first service pregnancy rate was noted (p < 0.01). The pregnancy rate was the lowest in the H protocol during warm period (p < 0.05). Treatment with hCG 5 days after AI significantly improved pregnancy rates in those cows that were treated with the H protocol compared with saline treatments (41.5% vs 24.8%; p < 0.01). O and H were more effective in primiparous than in pluriparous cows (46.1% vs 29.9%; p < 0.1 and 43.6% vs 24.6%; p < 0.01). First service pregnancy rates were higher in primiparous hCG-treated than in pluriparous hCG-treated cows (57.9% vs 32.3%; p < 0.01). The pregnancy rate was higher for the hCG-treated cows compared with saline-treated cows during warm period (37.9% vs 23.6%; p < 0.001). PMID- 19309468 TI - Does a febrile reaction to platelets predispose recipients to red blood cell alloimmunization? AB - BACKGROUND: A variable effect of inflammation on alloimmunization to transfused red blood cells (RBCs) in mice has been recently reported. We investigated whether RBC alloimmunization in humans was affected by transfusion of blood products in temporal proximity to experiencing a febrile transfusion reaction (FTR) to platelets (PLTs), an event predominantly mediated by inflammatory cytokines. STUDY DESIGN AND METHODS: Blood bank databases were used to identify patients who experienced an FTR or possible FTR to PLTs from August 2000 to March 2008 (FTR group). The control group of patients received a PLT transfusion on randomly selected dates without experiencing an FTR. The "event" was defined as the PLT transfusion that caused the FTR in the FTR group or the index PLT transfusion in the control group. The number of transfused blood products and their proximity to the event were recorded along with other recipient data. The primary endpoint was the rate of RBC alloimmunization between the two groups. RESULTS: There were 190 recipients in the FTR group and 245 in the control group. Overall, the recipients in the control group were younger and received more blood products on the day of their event and over the subsequent 10 days. The alloimmunization rate among recipients in the FTR group was higher than in the control group (8% vs. 3%, respectively; p = 0.026). CONCLUSIONS: These preliminary data support our hypothesis that recipient inflammation may affect RBC alloimmunization in humans; however, a more detailed understanding of the pathophysiologic association between inflammation and alloimmunization is required before definitive conclusions can be reached. PMID- 19309469 TI - Dead man talking: sustained utility of data from archaic paternity tests. AB - BACKGROUND: Almost all relationship analysts now use molecular (DNA) tests to obtain necessary genetic information, yet older blood group tests remain valid. Circumstances may provide blood test results from old reports to avoid trying to sample DNA from inaccessible sources. CASE STUDY: A mother recently claimed that a deceased man (alleged father [AF]) sired her child. Insurance investigation recovered two paternity test reports from an AABB-accredited laboratory. The 16- and 18-year-old reports employed blood groups and protein polymorphisms to exonerate two different men. One report contained the multilocus phenotypes of the AF and the other contained the phenotypes of the mother and child at the same loci. A new case was synthesized from the old reports. RESULTS: Genetic inconsistencies (three direct and one indirect) were demonstrated among the nine loci that had been typed in both the AF and the mother-child pair. CONCLUSION: New relationship tests may be reconstructed from phenotypes reported before the molecular test era. This approach avoids exhumation and other problematic methods of specimen retrieval. PMID- 19309470 TI - Blood use in the ambulatory setting among elderly in the United States. AB - BACKGROUND: Our study characterizes blood use in the ambulatory setting by US elderly Medicare beneficiaries during 2001. As the US population ages and delivery of health care in outpatient settings is on the rise, ambulatory blood utilization is expected to increase. There is currently a lack of broad population-based studies detailing ambulatory blood utilization patterns among the US elderly. STUDY DESIGN AND METHODS: This descriptive cross-sectional study of ambulatory blood utilization in institutional outpatient settings used Medicare administrative data (5% sample of enrollees) for calendar year 2001. Blood use was identified by either the presence of recorded blood units or the procedure code(s) for transfusion of whole blood or red blood cells. RESULTS: Among 1,368,368 elderly Medicare beneficiaries analyzed, 7054 (0.52%) had blood transfusion in institutional outpatient settings, and 34,186 (2.50%) had blood transfusion in the inpatient setting. Of 10,705 institutional outpatient claims with blood use quantified in this cohort of elderly Medicare beneficiaries, the top 10 principal diagnoses using the largest quantities of blood accounted for 66.3% of total blood units transfused. Nine of these 10 principal diagnoses were either for anemias (51%) or neoplasms (13%) and accounted for 64% of total blood units transfused. CONCLUSION: Our study suggests that most of the ambulatory blood utilization among US elderly is for diagnoses of anemias and neoplasms rather than procedures. Our population-based study provides valuable information on ambulatory blood utilization patterns which may be used to better understand the reasons for transfusion in the ambulatory setting as blood use is expected to grow. PMID- 19309471 TI - Cellular immune response to hepatitis C virus (HCV) in nonviremic blood donors with indeterminate anti-HCV reactivity. AB - BACKGROUND: Blood donors with indeterminate hepatitis C virus antibody (anti-HCV) reactivity are rejected from blood donation. As they are mostly nonviremic, the source of these reactions remains unclear. Reasons for such findings can be resolved HCV infections as well as unspecific antibody reactions. The aim of this study was to investigate HCV-specific T-cell response in blood donors to determine the reason for the weak antibody detection. STUDY DESIGN AND METHODS: Anti-HCV reactivity was tested in 72 blood donors initially diagnosed with an indeterminate HCV result by enzyme-linked immunosorbent assay and immunoblot. Cellular immune response was measured by proliferation assay and enzyme-linked immunosorbent spot analysis after stimulation with viral proteins core, NS3, and NS4. RESULTS: In 56% of donors anti-HCV reactivity was detectable in the screening assay whereas 72% had a reaction in the confirmation immunoblot. Forty six percent of donors had a cellular immune response against HCV proteins. The response was most frequent to NS3 protein. CONCLUSION: In almost half of donors the indeterminate result in serologic testings could be explained by a previous resolved HCV infection as the pattern of T-cell response was similar to these patients. These findings indicate that HCV-specific antibodies disappear more rapidly after resolved infection than HCV-specific T cells. These results are important for counseling blood donors and patients with indeterminate serologic results. PMID- 19309472 TI - A modeling framework for evaluation and comparison of trigger strategies for switching from minipool to individual-donation testing for West Nile virus. AB - BACKGROUND: To decrease the likelihood of transmission from donations containing West Nile virus (WNV) levels below minipool nucleic acid test (MP-NAT) detection limits, blood centers switch from MP-NAT to individual-donation testing (ID-NAT) after detection of MP-NAT-positive donations. The effectiveness of strategies to trigger or discontinue ID-NAT screening is largely unknown. STUDY DESIGN AND METHODS: Twenty-seven strategies to trigger and discontinue ID-NAT screening were evaluated with a statistical model based on known dynamics of WNV infection and historical data on WNV prevalence among blood donations. Breakthroughs were defined as WNV immunoglobulin M antibody-negative, viremic (RNA-positive) donations that could only be identified by ID-NAT, but were screened by MP-NAT. Effectiveness (proportional reduction of breakthroughs relative to MP-NAT screening alone) and efficiency (absolute reduction of breakthroughs relative to the number of tests performed) were estimated by simulating donation years of varying outbreak severities over a range of blood collection frequencies. RESULTS: Most strategies were effective (>75% reduction in breakthroughs) when daily donations exceeded 560. In larger centers (1008 donations daily), effectiveness of trigger-on strategies based on absolute number of MP-NAT positive donations improved, but worsened for strategies using rate-based criteria. Effectiveness increased slightly by triggering on one MP-NAT-positive rather than two and increased substantially by increasing the duration from 7 to 14 days that no ID-NAT-positive donations are detected before resuming MP-NAT. CONCLUSION: Most trigger strategies become effective when test results from at least 560 donations daily are considered. A 14-day ID-NAT period may improve safety relative to the increase in the number of tests performed. PMID- 19309473 TI - Transfusion of platelet components prepared with photochemical pathogen inactivation treatment during a Chikungunya virus epidemic in Ile de La Reunion. AB - BACKGROUND: During the Chikungunya virus (CHIKV) epidemic on Ile de La Reunion, France, more than 30% of 750,000 inhabitants were infected. Local blood donation was suspended to prevent transfusion-transmitted infection (TT-CHIKV). To sustain the availability of platelet (PLT) components, the Etablissement Francais du Sang implemented universal pathogen inactivation (INTERCEPT, Cerus Europe BV) of PLT components (CPAs). The study assessed the safety of PLT components treated with pathogen inactivation transfused in routine clinical practice. STUDY DESIGN AND METHODS: This was a retrospective observational study using patient medical records and the AFSSAPS hemovigilance database (eFIT) to identify TT-CHIKV and adverse events (AEs) classified as acute transfusion reactions (ATRs) to PLT components prepared with pathogen inactivation. RESULTS: During 1 year, 1950 INTERCEPT-CPAs were transfused to 335 adult, 51 pediatric, and 41 infant patients. Nineteen AEs were observed in 15 patients and 10 were classified as ATRs. Eight ATRs occurred in 6 pediatric hematology-oncology patients. No ATRs were observed in infants. The most frequently reported signs and symptoms were Grade 1 urticaria, itching, chills, fever, and anxiety. No cases of transfusion related acute lung injury, TT-sepsis, or TT-CHIKV were detected. CONCLUSIONS: INTERCEPT-CPAs were well tolerated in a broad range of patients, including infants. ATR incidence was low and when present ATRs were of mild severity. PMID- 19309474 TI - Impact of individual-donation nucleic acid testing on risk of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus transmission by blood transfusion in South Africa. AB - BACKGROUND: In 2005, the South African National Blood Service introduced individual-donation (ID) nucleic acid test (NAT) screening for human immunodeficiency virus (HIV) RNA, hepatitis C virus (HCV) RNA, and hepatitis B virus (HBV) DNA. At the same time the use of ethnic origin to prioritize the transfusion of blood according to a hierarchy of residual risk was discontinued. STUDY DESIGN AND METHODS: ID-NAT (Ultrio on Procleix Tigris, Chiron) and serology (PRISM, Abbott) repeat test and confirmation testing algorithms were designed to enable differentiation between false-positive and true-NAT and -serology yields. After 1 year, the NAT and serology yield rates in first-time, lapsed, and repeat donors were analyzed and used to estimate the residual risk of HIV, HBV, and HCV infections by blood transfusion. RESULTS: The HIV, HBV, and HCV ID-NAT window phase yield rates in 732,250 blood donations were 1:45,765, 1:11,810, and 1:732,200, respectively. Seven of 16 HIV window phase donations with viral loads above 16,000 copies/mL were HIV p24 antigen enzyme-linked immunosorbent assay positive. PRISM detected anti-HIV and hepatitis B surface antigen (HBsAg) in 89.4 and 73.9% of early infections in repeat donors. The Procleix assay detected viremia in 99.7 and 95.5% of anti-HIV- and HBsAg-positive first-time donors. In these donors, the occult HBV DNA carrier rate was 1:5200. The residual transmission risk of ID-NAT HIV, HBV, and HCV window phase donations was estimated at 1:479,000, 1:61,500, and 1:21,000,000 respectively. CONCLUSION: One year ID-NAT screening of 732,250 donations interdicted 16 HIV, 20 HBV, and 1 HCV window phase donations and 42 anti-hepatitis B core antigen-reactive infections during an early recovery or a later stage of occult HBV infection. PMID- 19309475 TI - Exchange versus simple transfusion for acute chest syndrome in sickle cell anemia adults. AB - BACKGROUND: There are scant data regarding the relative efficacy of exchange transfusion (XC) versus simple transfusion (ST) for treatment of sickle cell anemia acute chest syndrome (ACS). STUDY DESIGN AND METHODS: Twenty patients who received XC for ACS were compared with 20 ST patients. Hemoglobin (Hb) levels, platelet and white blood cell counts, lactate dehydrogenase (LDH), indirect bilirubin, and temperature were used to assess disease severity. Primary outcome was postprocedure length of hospital stay; secondary outcome was total length of stay. RESULTS: Cohorts were similar with regard to age; sex; prior ACS episodes; echocardiogram results; and antibiotic, bronchodilator, and hydroxyurea use. Maximum temperature recorded was higher in the XC group (39.1 degrees C vs. 38.4 degrees C, p = 0.02), but LDH, WBCs, and indirect bilirubin were comparable. Admission Hb levels were higher for XC (XC 8.6 g/dL vs. ST 7.4 g/dL, p = 0.02) and XC had higher peak Hb levels during hospitalization (10.4 +/- 1.4 g/dL vs. 9.3 +/- 1.0 g/dL, p < or = 0.01). No differences were demonstrable in postprocedure length of stay (XC 5.6 days vs. ST 5.9 days, p = 0.82) or total length of stay (XC 8.4 days vs. ST 8.0 days, p = 0.76). A total of 10.3 +/- 3.0 units were transfused for XC compared to 2.4 +/- 1.2 units for ST (p < 0.001). CONCLUSIONS: Based on postprocedure length of stay or total length of stay, we could not detect a difference in the efficacy of XC compared to ST in populations despite red blood cell product usage fourfold higher in the XC group. We suggest that it is time for an adequately powered, randomized trial to examine the true risk:benefit ratio of XC in ACS. PMID- 19309476 TI - D variants at the RhD vestibule in the weak D type 4 and Eurasian D clusters. AB - BACKGROUND: One branch of the RHD phylogenetic tree is represented by the weak D type 4 cluster of alleles with F223V as the primordial amino acid substitution. F223V as well as a large number of further substitutions causing D variants are located at the extracellular RhD protein vestibule, which represents the entrance to the transmembraneous channel of the RhD protein. STUDY DESIGN AND METHODS: RHD and RHCE nucleotide sequences were determined from genomic DNA and cDNA. D epitope patterns were established with commercial monoclonal anti-D panels. RESULTS: The RHD alleles DOL-1 and DOL-2 had the two amino acid substitutions M170T (509T>C) and F223V (667T>G) in common. DOL-2 harbored the additional substitution L378V (1132C>G). Both alleles were observed in Africans and are probably evolutionary related. DMI carried M170I (510G>A), which differed from the DOL-typical substitution. DFW and DFL harbored the substitutions H166P (497A>C) and Y165C (494A>G). The antigen densities of DOL-1, DFL, and DFW were only moderately reduced. CONCLUSION: DOL-1 and DOL-2 belong to the weak D type 4 cluster of RHD alleles. Together with DMI, DFL, and DFW they represent D variants with amino acid substitutions located at extracellular loops 3 or 4 lining the RhD protein vestibule. These substitutions were of minor influence on antigen density while adjacent substitutions in the transmembraneous section caused weak D antigen expression. All these D variants were partial D and alloanti-D immunizations have been observed in DOL-1, DMI, and DFL carriers. The substitution at position 170 causes partial D although located deep in the vestibule. PMID- 19309477 TI - A rare case of functional pancreas graft with newly developed collateral venous drainage after complete portal thrombosis. PMID- 19309478 TI - Successful outcome of paediatric en bloc kidney transplantation from the youngest donation-after-cardiac-death donor in the United Kingdom. PMID- 19309479 TI - Serum lipid profile: a predictor of clinical outcome in dengue infection. AB - OBJECTIVES: To correlate serum lipoproteins levels with dengue severity, clinical outcome (patient survival), bleeding severity, capillary leakage, supportive care requirement and hospital stay duration. METHODS: Single centre, prospective study. SETTING: Tertiary care hospital. SUBJECTS: 50 (1 month-18 years) dengue fever cases and 50 age and sex matched non-febrile controls. Methods: patients were classified (WHO criteria) as dengue fever (DF), dengue haemorrhagic fever (DHF), dengue shock syndrome (DSS). Bleeding score; serum triglyceride, cholesterol, HDL, LDL and VLDL was determined. Statistical analysis was done using Chi-squared test with continuity correction, Unpaired t-test, ANOVA with post hoc test, Binary logistic and multinomial regression. RESULTS: 18 (36%) patients were DF, 19 (38%) DHF and 13 (26%) DSS. Lowest cholesterol, VLDL levels seen in DSS and highest in DF. Mean cholesterol level is significantly lower in expired patients and patients with third spacing. Severe bleeding significantly correlated with cholesterol level and hepatic dysfunction but not with platelet count or coagulopathy. Duration of intravenous fluid requirement and packed cell requirement negatively correlated with HDL and cholesterol levels. Fresh frozen plasma requirement negatively correlated with TG, HDL, VLDL and cholesterol levels. Platelet transfusion requirement and duration of hospital stay did not correlate with lipid levels. Intensive care and ventilator requirement negatively correlated with cholesterol level; inotrope requirement negatively correlated with HDL level. Ventilator requirement correlated negatively with TG levels also. CONCLUSIONS: Lipid profile changes accompany dengue infection, some of which may indicate severity and guide therapy. PMID- 19309480 TI - Genetic characterization of antibiotic resistance in enteropathogenic Escherichia coli carrying extended-spectrum beta-lactamases recovered from diarrhoeic rabbits. AB - A total of 52 Escherichia coli strains isolated from diarrhoeic rabbits were investigated for their enteropathogenic E. coli (EPEC) pathotype by PCR amplification of eae and bfp virulence genes. A total of 22 EPEC isolates were identified, serotyped and studied for antibiotic resistance and screened for the detection of extended-spectrum beta-lactamases (ESBLs). The EPEC isolates belonged to three serogroups (O26, O92 and O103). The most common serogroup (O103:K-:H2) was observed among 17 EPEC strains, the O92:K-serogroup in three isolates (the antibiotic sensitive ones) and the remaining O26:K-serogroup in two isolates (the ESBLs isolates). Resistances to ampicillin and tetracycline were the most frequent and detected followed by resistance to nalidixic acid, streptomycin, trimethoprim-sulphamethoxazole, cefoxitin, gentamicin and ciprofloxacin. All the isolates were sensitive for amikacin, ceftazidime, aztreonam, imipenem, chloramphenicol, tobramycin and amoxicillin + clavulanic acid. Two isolates recovered from two adult animals showed an intermediate susceptibility to cefotaxime, and a positive screening test for ESBL was demonstrated in both. The bla(TEM) gene was demonstrated in the majority of ampicillin-resistant isolates. The aac(3)-II or aac(3)-IV genes were detected in the four gentamicin-resistant isolates. In addition, the aadA gene was detected in 60% of streptomycin-resistant isolates. The tet(A) or tet(B) genes were identified in all tetracycline-resistant isolates. A total of nine EPEC isolates showed the phenotype SXT-resistant, and the sul1 and/or sul2 and/or sul3 genes were detected in all of them. Our findings showed that the molecular detection by the eae and bfp genes by PCR followed by serotyping is useful for monitoring trends in EPEC infections of rabbits allowing the identification of their possible reservoirs. The detection of genes involved in the resistance to antibiotics of different families in a relatively high proportion of faecal E. coli isolates of rabbits is of great interest and could be considered a serious public health problem. PMID- 19309481 TI - Characterization of Staphylococcus aureus isolated from dairy animals in Ireland. AB - Enterotoxigenic Staphylococcus aureus frequently contaminate milk and milk products causing food poisoning. Staphylococcus aureus were isolated from bovine, ovine and caprine milk and milk filters from 78 dairy production holdings supplying the farmhouse cheese sector in Ireland, using standard culture methods. Molecular methods were applied to study the distribution of genes encoding staphylococcal enterotoxins and toxic shock syndrome toxin in the collection. Multilocus variable number tandem repeat analysis was used to subtype the collection. One hundred and two Staphylococcus aureus (54 milk filters and 48 bulk milk) were recovered from apparently healthy animals; half of the isolates were toxigenic. Our findings are discussed in light of the risks posed to public health. PMID- 19309482 TI - Prevalence of mastitis and brucellosis in cattle in Awassa and the peri-urban areas of two smaller towns. AB - The prevalence of mastitis and brucellosis in urban and peri-urban settings was studied in Awassa and two smaller nearby towns in southern Ethiopia, because milk born diseases are causing a risk for human health, besides direct impacts on animal production. Mastitis was investigated by examining 80 cows (320 udder quarters) using California mastitis test (CMT) and somatic cell count (SCC). The prevalence of brucellosis was assessed by sampling 177 cattle in Awassa and its peri-urban areas using serological methods. Logistic regression was used to analyse risk factors associated with mastitis. Prevalence of clinical mastitis on quarter level was 0.9%, and 1.9% of quarters were non-functional or blocked. Prevalence of sub-clinical mastitis at quarter level in urban and peri-urban areas was significantly different (P < 0.05). Cows in large herds and at advanced lactation number were associated with higher risk of infection. The percentage of quarters positive on CMT (42.5%) was close to the percentage-positive detected by SCC (41.2%). Prevalence of brucellosis was 3.9% in the peri-urban area, while no brucellosis cases were detected in Awassa. More frequent use of artificial insemination in the urban than in peri-urban area might have contributed to the absence of brucellosis in the urban location. The extent of mastitis is, however, a threat to the dairy enterprise in and around Awassa. Pasteurization of milk and milk products is indicated in some parts of the area because of the danger of brucellosis. PMID- 19309483 TI - Streptococcal cellulitis following preparation of fresh raw seafood. AB - We describe three zoonotic streptococcal soft tissue infections resulting from fresh seafood contact. One was a localized thumb infection with Streptococcus iniae in an immunocompetent healthy young male resulting from a puncture wound from a crab pincer. The other two were cases of ascending upper limb cellulitis associated with bacteraemia in mastectomy patients. One of these infections was caused by S. iniae while the other was caused by Streptococcus dysgalactiae subsp. dysgalactiae, a species that has not been previously described as a cause of zoonotic infection. Hence when cleaning raw seafood, protective equipment should be used to minimize the risk of percutaneous injuries. PMID- 19309484 TI - Rabies control in Finland: a 12-year experience of human and veterinary surveillance. AB - Little is known about the public health burden of rabies in rabies-free countries. In these countries, the surveillance of suspected and treated cases serves as a substitute for estimating the risk and burden of human rabies because deaths due to rabies are extremely rare. Suspected rabies exposures among Finnish inhabitants were characterized using data from the National Infectious Disease Registry as well as animal surveillance data from the Finnish Food Safety Authority Evira, 1995-2006. In total, 195 suspected rabies exposures were reported (incidence 3/million inhabitants/year). Exposures were equally common among both genders and the median age was 35 years. Exposures were more common among 20- to 49-year olds than among other age groups. Less than one-third of the exposures occurred in Finland (incidence of indigenous exposures 0.9/million inhabitants/year). Indigenous rabies exposures were most frequently reported in southeastern Finland, with cats and dogs as the main sources. The high prevalence in the Baltic countries and Russia poses a risk for rabies reintroduction. The present control of wildlife rabies appears successful and important. The import of animals from endemic areas, however, remains a risk, which can be reduced by increasing public awareness of the disease, vaccination of imported animals and better rabies control in endemic countries. PMID- 19309485 TI - Development and evaluation of a new growth medium for Helicobacter pylori. AB - The present study was aimed at modifying the original formulation of Commercial Eugon agar (CEA) to develop a new H. pylori growth medium. Initial studies were carried out to determine the number of H. pylori colonies recovered on in-house H. pylori agar (IHPA), IHPA without L-cysteine and sodium sulfite (IHPA-NC), IHPA without L-cysteine (IHPA-C), IHPA without sodium sulfite (IHPA-N) and CEA as the control. Significant differences (P < 0.001) in the number of colonies recovered were observed between IHPA-N, IHPA-NC and IHPA-C. Incorporation of sodium sulfite decreased the number of colonies recovered, indicating that sodium sulfite was inhibitory to H. pylori growth. Removal of L-cysteine reduced the number of colonies recovered, suggesting that L-cysteine is necessary for the growth of H. pylori. In the subsequent study, incorporation of K(2)HPO(4) further increased the number of colonies recovered compared with IHPA-N (P < 0.001), and 0.25% (w/v) of K(2)HPO(4) yielded the highest numbers of colonies (P < or = 0.04). Finally, thirty other H. pylori clinical isolates were evaluated for their growth in the IHPAP-N, a new medium consisting of 1.5% (w/v) pepticase, 0.5% (w/v) peptone, 0.4% (w/v) sodium chloride, 0.03% (w/v) L-cysteine, 0.55% (w/v) dextrose, 0.25% (w/v) K(2)HPO(4) and 1.5% (w/v) agar. The number of colonies recovered in IHPAP-N was significantly (P < 0.005) higher than that of CEA. IHPAP N with 0.25% K(2)HPO(4) and without sodium sulfite were adequate solid media for the growth of H. pylori. PMID- 19309486 TI - Asymptomatic healthy slaughterhouse workers in South Korea carrying Shiga toxin producing Escherichia coli. AB - A total of 1602 stool samples from healthy employees in a slaughter company were screened by PCR for Shiga toxin (Stx)-producing Escherichia coli (STEC). The PCR product of Stx-encoding genes was detected in 90 (5.6%) of 1602 stool samples. Among the 90 stx-positive workers, the Residual Products Handlers and Slaughterers had rates of 8.0% and 6.0%--higher than Inspectors, Grading Testers and Livestock Hygiene Controllers at 3.3%, 2.0% and 3.5%, respectively. Forty nine (54.4%) were shown to have stx2; 25 (27.7%) carried stx1 and 16 (17.7%) had both stx1 and 2. Distribution of the stx PCR-positive workers by age revealed an increase in STEC infection with age (P<0.05). Phenotypic and genotypic traits of nine STEC strains isolated from eight slaughter plant workers were characterized. A variety of serotypes, five O serogroups (O8, O54, O59, O103 and O153) and two H serogroups (H7 and H32) were found, but none of the strains belonged to the serogroup O157. Eight Vero cell cytotoxicity assay-positive strains were isolated from the workers and these workers were asymptomatic and healthy. The results of the study show that slaughter plant workers are at high risk of STEC infection. PMID- 19309487 TI - Recognition between tRNASer and archaeal seryl-tRNA synthetases monitored by suppression of bacterial amber mutations. AB - Two dissimilar seryl-tRNA synthetases (SerRSs) exist in Methanosarcina barkeri: one of bacterial type (bMbSerRS) and the other resembling SerRSs present only in methanogenic archaea (mMbSerRS). While the expression of the archaeal bMbSerRS gene in Escherichia coli complements the function of thermolabile SerRS at a nonpermissive temperature, mMbSerRS does not. Our recent X-ray structural analysis of mMbSerRS revealed an idiosyncratic N-terminal domain and a catalytic zinc ion in the active site, identifying methanogenic-type SerRSs as atypical members of the SerRS family. To shed further light on substrate discrimination by methanogenic-type SerRS, we developed an in vivo system in E. coli to study tRNA serylation by mMbSerRS variants. We show that coexpression of the M. barkeri SerRS gene, encoding either bacterial- or methanogenic-type SerRS, with the gene for cognate archaeal suppressor tRNA leads to suppression of bacterial amber mutations, implying that the E. coli translation machinery can use serylated tRNA from methanogenic archaea as a substrate in protein synthesis. Furthermore, because serylation of M. barkeri serine-specific tRNA by endogenous E. coli SerRS is negligible, suppression is entirely dependent on recognition between archaeal partners (mMbSerRS/suppressor tRNA(Ser)). Thus, the efficiency of suppression by mMbSerRS variants quantified in the described beta-galactosidase-based reporter system, accurately reflects enzymes' serylation propensity obtained by in vitro kinetic measurements. PMID- 19309488 TI - Patients' perceptions and experiences of transitions in diabetes care: a longitudinal qualitative study. AB - OBJECTIVE: To examine patients' perceptions and experiences over time of the devolvement of diabetes care/reviews from secondary to primary health-care settings. DESIGN: Repeat in-depth interviews with 20 patients over 4 years. PARTICIPANTS AND SETTING: Twenty type 2 diabetes patients recruited from primary- and secondary-care settings across Lothian, Scotland. RESULTS: Patients' views about their current diabetes care were informed by their previous service contact. The devolvement of diabetes care/reviews to general practice was presented as a 'mixed blessing'. Patients gained reassurance from their perception that receiving practice-based care/reviews signified that their diabetes was well-controlled. However, they also expressed resentment that, by achieving good control, they received what they saw as inferior care and/or less frequent reviews to others with poorer control. While patients tended to regard GPs as having adequate expertise to conduct their practice-based reviews, they were more ambivalent about nurses taking on this role. Opportunities to receive holistic care in general practice were not always realized due to patients seeing health-care professionals for diabetes management to whom they would not normally present for other health issues. CONCLUSIONS: It is important to educate patients about their care pathways, and to reassure them that frequency of reviews depends more on clinical need than location of care and that similar care guidelines are followed in hospital clinics and general practice. A patients' history of service contact may need to be taken into account in future studies of service satisfaction. PMID- 19309489 TI - Overexpression of chromatin assembly factor-1 (CAF-1) p60 is predictive of adverse behaviour of prostatic cancer. AB - AIMS: Prostatic cancer may remain organ-confined indefinitely; in a number of patients, however it gives rise to clinical symptoms and death. The biological behaviour of this tumour mostly remains difficult to predict. A promising tool for diagnosis and prognosis of some human tumours is the chromatin assembly factor-1 (CAF-1), involved in the control of higher order chromatin organization. The aim was to explore the role of CAF-1/p60 protein as a new prognostic marker for prostatic cancer. METHODS AND RESULTS: The expression of CAF-1/p60 was evaluated by immunohistochemistry and Western blotting in a selected series of prostatic cancers and in prostatic cancer cell lines. Results were compared with clinicopathological data and outcome of patients. CAF-1/p60 was expressed in all cases, with a linear increase from low-grade tumours (Gleason score <7) to high grade prostatic cancers (Gleason score >7). By comparing results with follow-up data, a significant association between overexpression of CAF-1/p60 and unfavourable behaviour of prostatic cancer emerged, and its predictive value was independent of classical prognostic factors. CONCLUSIONS: In our series of cases, overexpression of CAF-1/p60 characterized prostatic cancers with a worse prognosis. CAF-1/p60 has a potential role as a new reliable prognostic biomarker for prostatic cancer. PMID- 19309490 TI - Nuclear, compared with combined nuclear and cytoplasmic expression of maspin, is linked in lung adenocarcinoma to reduced VEGF-A levels and in Stage I, improved survival. AB - AIMS: To evaluate whether there is a correlation between the subcellular localization of maspin and the histological, molecular and biological features of pulmonary adenocarcinoma, particularly addressing the hypothesis that the tumour inhibitor properties of maspin may be linked to a nuclear, compared with a combined nuclear and cytoplasmic expression pattern. METHODS AND RESULTS: The subcellular expression of maspin was determined in 80 resected pulmonary adenocarcinomas (Stage I, 46; Stage II, 10; Stage III, 20; Stage IV, 4) and correlated with histological grade, proliferative rate, p53 expression, vascular endothelial growth factor (VEGF)-A levels, and prognosis (mean follow-up of 41.5 months). Cases with nuclear (N) maspin (n = 47), compared with the [N + cytoplasmic (C)] group (n = 28), showed lower (P 50% having no predicted function. A small scale RNA interference (RNAi) screen was carried out in Trypanosoma brucei to evaluate the prospects for identifying novel cycle regulators. RESULTS: Procyclic form T. brucei were transfected with a genomic RNAi library and 204 clones isolated. However, only 76 RNAi clones were found to target a protein coding gene of potential interest. These clones were screened for defects in proliferation and cell cycle progression following RNAi induction. Sixteen clones exhibited proliferation defects upon RNAi induction, with eight clones displaying potential cell cycle defects. To confirm the phenotypes, new RNAi cell lines were generated and characterised for five genes targeted in these clones. While we confirmed that the targeted genes are essential for proliferation, we were unable to unambiguously classify them as cell cycle regulators. CONCLUSION: Our study identified genes essential for proliferation, but did not, as hoped, identify novel cell cycle regulators. Screening of the RNAi library for essential genes was extremely labour-intensive, which was compounded by the suboptimal quality of the library. For such a screening method to be viable for a large scale or genome wide screen, a new, significantly improved RNAi library will be required, and automated phenotyping approaches will need to be incorporated. PMID- 19309511 TI - The effect of long-term unilateral deafness on the activation pattern in the auditory cortices of French-native speakers: influence of deafness side. AB - BACKGROUND: In normal-hearing subjects, monaural stimulation produces a normal pattern of asynchrony and asymmetry over the auditory cortices in favour of the contralateral temporal lobe. While late onset unilateral deafness has been reported to change this pattern, the exact influence of the side of deafness on central auditory plasticity still remains unclear. The present study aimed at assessing whether left-sided and right-sided deafness had differential effects on the characteristics of neurophysiological responses over auditory areas. Eighteen unilaterally deaf and 16 normal hearing right-handed subjects participated. All unilaterally deaf subjects had post-lingual deafness. Long latency auditory evoked potentials (late-AEPs) were elicited by two types of stimuli, non-speech (1 kHz tone-burst) and speech-sounds (voiceless syllable/pa/) delivered to the intact ear at 50 dB SL. The latencies and amplitudes of the early exogenous components (N100 and P150) were measured using temporal scalp electrodes. RESULTS: Subjects with left-sided deafness showed major neurophysiological changes, in the form of a more symmetrical activation pattern over auditory areas in response to non-speech sound and even a significant reversal of the activation pattern in favour of the cortex ipsilateral to the stimulation in response to speech sound. This was observed not only for AEP amplitudes but also for AEP time course. In contrast, no significant changes were reported for late-AEP responses in subjects with right-sided deafness. CONCLUSION: The results show that cortical reorganization induced by unilateral deafness mainly occurs in subjects with left sided deafness. This suggests that anatomical and functional plastic changes are more likely to occur in the right than in the left auditory cortex. The possible perceptual correlates of such neurophysiological changes are discussed. PMID- 19309512 TI - Effect of a printed reminder in the waiting room to turn off mobile phones during consultation: a before and after study. AB - BACKGROUND: Telephone interruptions during consultations are one cause of work related stress amongst general practitioners. Many health care centers recommend that patients turn off any mobile phones to avoid interruptions to the discussion with the physicians. METHODS: The purpose of this before and after study was to determine whether a printed reminder for turning off the mobile phone in the waiting room is helpful in decreasing the number of interruptions during consultation. A visual phone off sign utilizing the International "No" symbol of a diagonal line through a circle, along with a "please turn off your phone during consultation" reminder was used in the waiting room in the "after" period. RESULTS: A significant difference was found in the proportion of patients receiving or making a call during the consultation (8.8% vs. 13.5%, RR = 0.66; 95%CI 0.46-0.94; p = 0.021) and in the total number of calls (10.4% vs. 17.3%, RR = 0.60; 95%CI 0.44-0.83, p = 0.003) between the exposed and the non-exposed groups. However, no significant differences were found in the total time or the median time spent talking during consultation. The duration of the calls had median times of 20.5 seconds and 22.3 seconds in the exposed and the non-exposed groups respectively. Women from both groups who received a call during consultation answered significantly more when compared to men (70% vs. 52%; p = 0.05); CONCLUSION: Our findings suggest that a printed reminder in the waiting room is helpful in decreasing the number of interruptions by mobile phone during consultation in our settings. The study provides the basis for further quantitative and qualitative research on this topic. PMID- 19309513 TI - Identifying disease-specific genes based on their topological significance in protein networks. AB - BACKGROUND: The identification of key target nodes within complex molecular networks remains a common objective in scientific research. The results of pathway analyses are usually sets of fairly complex networks or functional processes that are deemed relevant to the condition represented by the molecular profile. To be useful in a research or clinical laboratory, the results need to be translated to the level of testable hypotheses about individual genes and proteins within the condition of interest. RESULTS: In this paper we describe novel computational methodology capable of predicting key regulatory genes and proteins in disease- and condition-specific biological networks. The algorithm builds shortest path network connecting condition-specific genes (e.g. differentially expressed genes) using global database of protein interactions from MetaCore. We evaluate the number of all paths traversing each node in the shortest path network in relation to the total number of paths going via the same node in the global network. Using these numbers and the relative size of the initial data set, we determine the statistical significance of the network connectivity provided through each node. We applied this method to gene expression data from psoriasis patients and identified many confirmed biological targets of psoriasis and suggested several new targets. Using predicted regulatory nodes we were able to reconstruct disease pathways that are in excellent agreement with the current knowledge on the pathogenesis of psoriasis. CONCLUSION: The systematic and automated approach described in this paper is readily applicable to uncovering high-quality therapeutic targets, and holds great promise for developing network-based combinational treatment strategies for a wide range of diseases. PMID- 19309514 TI - A national clinical decision support infrastructure to enable the widespread and consistent practice of genomic and personalized medicine. AB - BACKGROUND: In recent years, the completion of the Human Genome Project and other rapid advances in genomics have led to increasing anticipation of an era of genomic and personalized medicine, in which an individual's health is optimized through the use of all available patient data, including data on the individual's genome and its downstream products. Genomic and personalized medicine could transform healthcare systems and catalyze significant reductions in morbidity, mortality, and overall healthcare costs. DISCUSSION: Critical to the achievement of more efficient and effective healthcare enabled by genomics is the establishment of a robust, nationwide clinical decision support infrastructure that assists clinicians in their use of genomic assays to guide disease prevention, diagnosis, and therapy. Requisite components of this infrastructure include the standardized representation of genomic and non-genomic patient data across health information systems; centrally managed repositories of computer processable medical knowledge; and standardized approaches for applying these knowledge resources against patient data to generate and deliver patient-specific care recommendations. Here, we provide recommendations for establishing a national decision support infrastructure for genomic and personalized medicine that fulfills these needs, leverages existing resources, and is aligned with the Roadmap for National Action on Clinical Decision Support commissioned by the U.S. Office of the National Coordinator for Health Information Technology. Critical to the establishment of this infrastructure will be strong leadership and substantial funding from the federal government. SUMMARY: A national clinical decision support infrastructure will be required for reaping the full benefits of genomic and personalized medicine. Essential components of this infrastructure include standards for data representation; centrally managed knowledge repositories; and standardized approaches for leveraging these knowledge repositories to generate patient-specific care recommendations at the point of care. PMID- 19309515 TI - Sexual differentiation of the zebra finch song system: potential roles for sex chromosome genes. AB - BACKGROUND: Recent evidence suggests that some sex differences in brain and behavior might result from direct genetic effects, and not solely the result of the organizational effects of steroid hormones. The present study examined the potential role for sex-biased gene expression during development of sexually dimorphic singing behavior and associated song nuclei in juvenile zebra finches. RESULTS: A microarray screen revealed more than 2400 putative genes (with a false discovery rate less than 0.05) exhibiting sex differences in the telencephalon of developing zebra finches. Increased expression in males was confirmed in 12 of 20 by qPCR using cDNA from the whole telencephalon; all of these appeared to be located on the Z sex chromosome. Six of the genes also showed increased expression in one or more of the song control nuclei of males at post-hatching day 25. Although the function of half of the genes is presently unknown, we have identified three as: 17-beta-hydroxysteroid dehydrogenase type IV, methylcrotonyl CoA carboxylase, and sorting nexin 2. CONCLUSION: The data suggest potential influences of these genes in song learning and/or masculinization of song system morphology, both of which are occurring at this developmental stage. PMID- 19309516 TI - Socioeconomic inequality in child injury in Bangladesh - implication for developing countries. AB - BACKGROUND: Child injury is an emerging public health issue in both developed and developing countries. It is the main cause of deaths and disabilities of children after infancy. The aim of this study was to investigate the socioeconomic inequality in injury related morbidity and mortality among 1-4 years children. MATERIALS AND METHODS: Data used for this study derived from Bangladesh Health and Injury Survey. A multistage cluster sampling technique was conducted for this survey. In this study quintiles of socioeconomic status were calculated on the basis of assets and wealth score by using principle component analysis. The numerical measures of inequality in mortality and morbidity were assessed by the concentration index. RESULTS: The poorest-richest quintile ratio of mortality due to injury was 6.0 whereas this ratio was 5.6 and 5.5 for the infectious diseases and non-communicable diseases. The values of mortality concentration indices for child mortality due to infection, non-communicable diseases and injury causes were -0.40, -0.32 and -0.26 respectively. Among the morbidity concentration indices, injury showed significantly greater inequality. All the concentration indices revealed that there were significant inequalities among the groups. The logistic regression analysis indicated that poor children were 2.8 times more likelihood to suffer from injury mortality than rich children, taking into account all the other factors. CONCLUSION: Despite concentration indices used in this study, the analysis reflected the family's socioeconomic position in a Bangladesh context, showing a very strong statistical association with child mortality. Due to the existing socioeconomic situation in Bangladesh, the poor children were more vulnerable to injury occurrence. PMID- 19309517 TI - Characterization of oxylipins and dioxygenase genes in the asexual fungus Aspergillus niger. AB - BACKGROUND: Aspergillus niger is an ascomycetous fungus that is known to reproduce through asexual spores, only. Interestingly, recent genome analysis of A. niger has revealed the presence of a full complement of functional genes related to sexual reproduction 1. An example of such genes are the dioxygenase genes which in Aspergillus nidulans, have been shown to be connected to oxylipin production and regulation of both sexual and asexual sporulation 234. Nevertheless, the presence of sex related genes alone does not confirm sexual sporulation in A. niger. RESULTS: The current study shows experimentally that A. niger produces the oxylipins 8,11-dihydroxy octadecadienoic acid (8,11-diHOD), 5,8-dihydroxy octadecadienoic acid (5,8-diHOD), lactonized 5,8-diHOD, 8-hydroxy octadecadienoic acid (8-HOD), 10-hydroxy octadecadienoic acid (10-HOD), small amounts of 8-hydroxy octadecamonoenoic acid (8-HOM), 9-hydroxy octadecadienoic acid (9-HOD) and 13-hydroxy octadecadienoic acid (13-HOD). Importantly, this study shows that the A. niger genome contains three putative dioxygenase genes, ppoA, ppoC and ppoD. Expression analysis confirmed that all three genes are indeed expressed under the conditions tested. CONCLUSION: A. niger produces the same oxylipins and has similar dioxygenase genes as A. nidulans. Their presence could point towards the existence of sexual reproduction in A. niger or a broader role for the gene products in physiology, than just sexual development. PMID- 19309518 TI - Biocomputational prediction of non-coding RNAs in model cyanobacteria. AB - BACKGROUND: In bacteria, non-coding RNAs (ncRNA) are crucial regulators of gene expression, controlling various stress responses, virulence, and motility. Previous work revealed a relatively high number of ncRNAs in some marine cyanobacteria. However, for efficient genetic and biochemical analysis it would be desirable to identify a set of ncRNA candidate genes in model cyanobacteria that are easy to manipulate and for which extended mutant, transcriptomic and proteomic data sets are available. RESULTS: Here we have used comparative genome analysis for the biocomputational prediction of ncRNA genes and other sequence/structure-conserved elements in intergenic regions of the three unicellular model cyanobacteria Synechocystis PCC6803, Synechococcus elongatus PCC6301 and Thermosynechococcus elongatus BP1 plus the toxic Microcystis aeruginosa NIES843. The unfiltered numbers of predicted elements in these strains is 383, 168, 168, and 809, respectively, combined into 443 sequence clusters, whereas the numbers of individual elements with high support are 94, 56, 64, and 406, respectively. Removing also transposon-associated repeats, finally 78, 53, 42 and 168 sequences, respectively, are left belonging to 109 different clusters in the data set. Experimental analysis of selected ncRNA candidates in Synechocystis PCC6803 validated new ncRNAs originating from the fabF-hoxH and apcC-prmA intergenic spacers and three highly expressed ncRNAs belonging to the Yfr2 family of ncRNAs. Yfr2a promoter-luxAB fusions confirmed a very strong activity of this promoter and indicated a stimulation of expression if the cultures were exposed to elevated light intensities. CONCLUSION: Comparison to entries in Rfam and experimental testing of selected ncRNA candidates in Synechocystis PCC6803 indicate a high reliability of the current prediction, despite some contamination by the high number of repetitive sequences in some of these species. In particular, we identified in the four species altogether 8 new ncRNA homologs belonging to the Yfr2 family of ncRNAs. Modelling of RNA secondary structures indicated two conserved single-stranded sequence motifs that might be involved in RNA-protein interactions or in the recognition of target RNAs. Since our analysis has been restricted to find ncRNA candidates with a reasonable high degree of conservation among these four cyanobacteria, there might be many more, requiring direct experimental approaches for their identification. PMID- 19309519 TI - Pathways to malaria persistence in remote central Vietnam: a mixed-method study of health care and the community. AB - BACKGROUND: There is increasing interest in underlying socio-cultural, economic, environmental and health-system influences on the persistence of malaria. Vietnam is a Mekong regional 'success story' after dramatic declines in malaria incidence following introduction of a national control program providing free bed-nets, diagnosis and treatment. Malaria has largely retreated to pockets near international borders in central Vietnam, where it remains a burden particularly among impoverished ethnic minorities. In these areas commune and village health workers are lynchpins of the program. This study in the central province of Quang Tri aimed to contribute to more effective malaria control in Vietnam by documenting the non-biological pathways to malaria persistence in two districts. METHODS: Multiple and mixed (qualitative and quantitative) methods were used. The formative stage comprised community meetings, observation of bed-net use, and focus group discussions and semi-structured interviews with health managers, providers and community. Formative results were used to guide development of tools for the assessment stage, which included a provider quiz, structured surveys with 160 community members and 16 village health workers, and quality check of microscopy facilities and health records at district and commune levels. Descriptive statistics and chi-square analysis were used for quantitative data. RESULTS: The study's key findings were the inadequacy of bed-nets (only 45% of households were fully covered) and sub-optimal diagnosis and treatment at local levels. Bed-net insufficiencies were exacerbated by customary sleeping patterns and population mobility. While care at district level seemed good, about a third of patients reportedly self-discharged early and many were lost to follow-up. Commune and village data suggested that approximately half of febrile patients were treated presumptively, and 10 village health workers did not carry artesunate to treat the potentially deadly and common P. falciparum malaria. Some staff lacked diagnostic skills, time for duties, and quality microscopy equipment. A few gaps were found in community knowledge and reported behaviours. CONCLUSION: Malaria control cannot be achieved through community education alone in this region. Whilst appropriate awareness-raising is needed, it is most urgent to address weaknesses at systems level, including bed-net distribution, health provider staffing and skills, as well as equipment and supplies. PMID- 19309520 TI - Nitric oxide synthase 2A (NOS2A) polymorphisms are not associated with invasive pneumococcal disease. AB - BACKGROUND: Streptococcus pneumoniae (pneumococcus) is responsible for over one million deaths per year, with young children, the elderly and immunocompromised individuals being most at risk. Approximately half of East African children have been reported to be asymptomatic carriers of pneumococcus with invasive infection occurring after the disruption of the respiratory membrane which is believed to be caused by the host immune response. Racial incidence of invasive pneumococcal disease (IPD) is higher in certain populations even after adjusting for environmental factors suggesting a genetic component to disease susceptibility. The nitric oxide synthase 2A (NOS2A) gene is responsible for the production of nitric oxide under pathological conditions including host defence against bacterial infection. Nitric oxide is a modulator of apoptotic and inflammatory cascades and endothelial permeability. We hypothesised that genetic variants within this gene may predispose to disease risk and survival. METHODS: A cohort of 299 children with IPD (221 meningitis, 41 pneumonia and 37 with bacteraemia) and 931 age matched controls from Malawi were used in this study. We investigated nine haplotype tagging single nucleotide polymorphisms within the NOS2A gene and compared the presence or absence of the minor alleles in cases and controls and survivors and non-survivors within the cases. RESULTS: We observed no significant associations between cases and controls or with survival in either all IPD cases or in the separate analysis of meningitis cases. A near significant association was obtained for the comparison of rs8078340 in cases and controls (p-value, 0.078). However, results were unadjusted for multiple testing. CONCLUSION: Our results suggest that polymorphic variation within the NOS2A gene does not influence invasive pneumococcal disease susceptibility or survival. PMID- 19309521 TI - Migration of Chadic speaking pastoralists within Africa based on population structure of Chad Basin and phylogeography of mitochondrial L3f haplogroup. AB - BACKGROUND: Chad Basin, lying within the bidirectional corridor of African Sahel, is one of the most populated places in Sub-Saharan Africa today. The origin of its settlement appears connected with Holocene climatic ameliorations (aquatic resources) that started ~10,000 years before present (YBP). Although both Nilo Saharan and Niger-Congo language families are encountered here, the most diversified group is the Chadic branch belonging to the Afro-Asiatic language phylum. In this article, we investigate the proposed ancient migration of Chadic pastoralists from Eastern Africa based on linguistic data and test for genetic traces of this migration in extant Chadic speaking populations. RESULTS: We performed whole mitochondrial genome sequencing of 16 L3f haplotypes, focused on clade L3f3 that occurs almost exclusively in Chadic speaking people living in the Chad Basin. These data supported the reconstruction of a L3f phylogenetic tree and calculation of times to the most recent common ancestor for all internal clades. A date ~8,000 YBP was estimated for the L3f3 sub-haplogroup, which is in good agreement with the supposed migration of Chadic speaking pastoralists and their linguistic differentiation from other Afro-Asiatic groups of East Africa. As a whole, the Afro-Asiatic language family presents low population structure, as 92.4% of mtDNA variation is found within populations and only 3.4% of variation can be attributed to diversity among language branches. The Chadic speaking populations form a relatively homogenous cluster, exhibiting lower diversification than the other Afro-Asiatic branches (Berber, Semitic and Cushitic). CONCLUSION: The results of our study support an East African origin of mitochondrial L3f3 clade that is present almost exclusively within Chadic speaking people living in Chad Basin. Whole genome sequence-based dates show that the ancestral haplogroup L3f must have emerged soon after the Out-of-Africa migration (around 57,100 +/- 9,400 YBP), but the "Chadic" L3f3 clade has much less internal variation, suggesting an expansion during the Holocene period about 8,000 +/- 2,500 YBP. This time period in the Chad Basin is known to have been particularly favourable for the expansion of pastoralists coming from northeastern Africa, as suggested by archaeological, linguistic and climatic data. PMID- 19309522 TI - Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma. AB - BACKGROUND: HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas. METHODS: To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH. RESULTS: The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+ immunostaining cases, which showed high-level amplification (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and HER-2 downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350). CONCLUSION: Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene. PMID- 19309523 TI - R-CHOP versus R-CVP in the treatment of follicular lymphoma: a meta-analysis and critical appraisal of current literature. AB - PURPOSE: R-CHOP (rituximab with cyclophosphamide, doxorubicin, vincristine and prednisone) and R-CVP (rituximab with cyclophosphamide, vincristine and prednisone) have both been used successfully in the treatment of patients with symptomatic follicular lymphoma (FL). No study has compared the efficacy of the two treatment modalities and attempted to evaluate the role of anthracyclines in the management of patients with FL. We conducted a meta-analysis of relevant literature comparing the two treatment arms for FL with response being the final endpoint. PATIENTS AND METHODS: TWO ANALYSES WERE CONDUCTED: The first analysis compared R-CHOP to R-CVP as frontline agents for the treatment of FL, and the second analysis included both untreated and relapsed patients. RESULTS: For both studies, R-CVP was superior to R-CHOP when evaluating for complete response (CR). Odds ratios were 2.86 (95% CI, 1.81-4.51) in the first analysis and 1.48 (95% CI, 0.991-2.22) in the second analysis. However for overall response (CR+Partial response, PR), R-CHOP was superior, with odds ratios of 5.45 (95% CI: 2.51 - 11.83) and 5.54 (95% CI: 2.69 - 11.40), for the first and second analyses, respectively. CONCLUSION: R-CHOP and R-CVP protocols achieve excellent overall response. In patients with known cardiac history, omission of anthracyclines is reasonable and R-CVP provides a competitive CR rate. In younger patients with FL where cumulative cardio-toxicity may be of importance in the long term and in whom future stem cell transplantation is an option, again R-CVP may be a more appealing option. PMID- 19309524 TI - Utility of EST-derived SSR in cultivated peanut (Arachis hypogaea L.) and Arachis wild species. AB - BACKGROUND: Lack of sufficient molecular markers hinders current genetic research in peanuts (Arachis hypogaea L.). It is necessary to develop more molecular markers for potential use in peanut genetic research. With the development of peanut EST projects, a vast amount of available EST sequence data has been generated. These data offered an opportunity to identify SSR in ESTs by data mining. RESULTS: In this study, we investigated 24,238 ESTs for the identification and development of SSR markers. In total, 881 SSRs were identified from 780 SSR-containing unique ESTs. On an average, one SSR was found per 7.3 kb of EST sequence with tri-nucleotide motifs (63.9%) being the most abundant followed by di- (32.7%), tetra- (1.7%), hexa- (1.0%) and penta-nucleotide (0.7%) repeat types. The top six motifs included AG/TC (27.7%), AAG/TTC (17.4%), AAT/TTA (11.9%), ACC/TGG (7.72%), ACT/TGA (7.26%) and AT/TA (6.3%). Based on the 780 SSR containing ESTs, a total of 290 primer pairs were successfully designed and used for validation of the amplification and assessment of the polymorphism among 22 genotypes of cultivated peanuts and 16 accessions of wild species. The results showed that 251 primer pairs yielded amplification products, of which 26 and 221 primer pairs exhibited polymorphism among the cultivated and wild species examined, respectively. Two to four alleles were found in cultivated peanuts, while 3-8 alleles presented in wild species. The apparent broad polymorphism was further confirmed by cloning and sequencing of amplified alleles. Sequence analysis of selected amplified alleles revealed that allelic diversity could be attributed mainly to differences in repeat type and length in the microsatellite regions. In addition, a few single base mutations were observed in the microsatellite flanking regions. CONCLUSION: This study gives an insight into the frequency, type and distribution of peanut EST-SSRs and demonstrates successful development of EST-SSR markers in cultivated peanut. These EST-SSR markers could enrich the current resource of molecular markers for the peanut community and would be useful for qualitative and quantitative trait mapping, marker-assisted selection, and genetic diversity studies in cultivated peanut as well as related Arachis species. All of the 251 working primer pairs with names, motifs, repeat types, primer sequences, and alleles tested in cultivated and wild species are listed in Additional File 1. PMID- 19309525 TI - Interaction of Cupidin/Homer2 with two actin cytoskeletal regulators, Cdc42 small GTPase and Drebrin, in dendritic spines. AB - BACKGROUND: Homer is a postsynaptic scaffold protein that links various synaptic signaling proteins, including the type I metabotropic glutamate receptor subunits 1alpha and 5, the inositol 1,4,5-trisphosphate receptor, Shank and Cdc42 small GTPase. Overexpression of Homer induces changes in dendritic spine morphology in cultured hippocampal neurons. However, the molecular basis underpinning Homer mediated spine morphogenesis remains unclear. In this study, we aimed to elucidate the structural and functional properties of the interaction between Cupidin/Homer2 and two actin-cytoskeletal regulators, Cdc42 small GTPase and Drebrin. RESULTS: Cupidin/Homer2 interacted with activated Cdc42 small GTPase via the Cdc42-binding domain that resides around amino acid residues 191-283, within the C-terminal coiled-coil domain. We generated a Cupidin deletion mutant lacking amino acids 191-230 (CPDDelta191-230), which showed decrease Cdc42-binding ability but maintained self-multimerization ability. Cupidin suppressed Cdc42 induced filopodia-like protrusion formation in HeLa cells, whereas CPDDelta191 230 failed to do so. In cultured hippocampal neurons, Cupidin was targeted to dendritic spines, whereas CPDDelta191-230 was distributed in dendritic shafts as well as spines. Overexpression of CPDDelta191-230 decreased the number of synapses and reduced the amplitudes of miniature excitatory postsynaptic currents in hippocampal neurons. Cupidin interacted with a dendritic spine F-actin-binding protein, Drebrin, which possesses two Homer ligand motifs, via the N-terminal EVH 1 domain. CPDDelta191-230 overexpression decreased Drebrin clustering in the dendritic spines of hippocampal neurons. CONCLUSION: These results indicate that Cupidin/Homer2 interacts with the dendritic spine actin regulators Cdc42 and Drebrin via its C-terminal and N-terminal domains, respectively, and that it may be involved in spine morphology and synaptic properties. PMID- 19309526 TI - A novel c.-22T>C mutation in GALK1 promoter is associated with elevated galactokinase phenotype. AB - BACKGROUND: Many genetic variations of GALK1 have been identified in the patients with galactokinase (GALK1) deficiency. However, the molecular characteristics of GALK1 in individuals with elevated GALK1 activity are relatively unknown. METHODS: We investigated the relationship between elevated GALK1 activity and the molecular GALK1 gene variations, and the molecular mechanism underlying elevated GALK1 activity. PCR products from 63 subjects, without any attenuation of galactose degradation enzymes, were sequenced to screen for nucleotide alterations in the GALK1 promoter. RESULTS: Three nucleotide substitutions were identified: c.-179A>G, c.-27A>C, and c.-22T>C. With respect to the c.-22T>C mutation, GALK1 activity in 13 subjects with the T/C or C/C genotype was significantly higher than those in 50 subjects with the T/T genotype (p < 0.001). The dual luciferase reporter assay in Hep3B cells showed that the luciferase activity with the GALK1 promoter with the c.-22C mutant allele increased approximately 2.5-fold, compared to that with the c.-22T. A specific DNA-protein complex was observed in an electrophoretic mobility shift assay, with slightly higher affinity to c.-22C than to c.-22T. CONCLUSION: The c.-22T>C mutation, which was observed frequently in individuals with elevated GALK1 activity, increased the expression of a reporter gene through enhanced binding of a currently unidentified nuclear protein. These results suggest that the elevated GALK1 activity resulted from enhanced gene expression, due to nucleotide variation within GALK1 promoter. PMID- 19309527 TI - Determinants of maternal and umbilical blood lead levels: a cross-sectional study, Mosul, Iraq. AB - BACKGROUND: The populations who are most sensitive to lead exposure from various sources are pregnant women and their newborns. Aiming to explore the presence of correlation between maternal and cord blood lead levels and to identify potential predictors that may influence both levels, the present study has been conducted. METHODS: A cross-sectional study was conducted covering 350 full terms maternal newborns pairs from Mosul maternity hospitals. Data were obtained directly from women just before delivery by the use of a detailed questionnaire form.Maternal and umbilical blood lead levels were estimated using LEADCARE Blood Lead Testing System and Kits. RESULTS: A positive significant correlation was found between maternal and cord blood lead values (r = 0.856, p = 0.001). By backward stepwise logistic regression analysis the followings emerged as significant potential predictors of high maternal blood lead: low parity, smoking and Hb level <11 gm/dl. Regarding cord blood lead: coffee consumption and high maternal blood lead were significant risk predictors. Milk and milk products consumption, calcium intake and low level of physical activity were significantly operational in the prevention of high maternal blood lead levels. Iron intake and also low level of physical activity were shown as significant protective variables against high cord blood lead values. CONCLUSION: Study results have provided baseline data needed to be transformed to decision makers to implement measures to eliminate lead from the environment and protect future generation from its deleterious effects. PMID- 19309528 TI - Specific and complete human genome amplification with improved yield achieved by phi29 DNA polymerase and a novel primer at elevated temperature. AB - BACKGROUNDS: Whole genome amplification (WGA) is a practical solution to eliminate molecular analysis limitations associated with genomic DNA (gDNA) quantity. Different methods have been developed to amplify the whole genome, including primer extension preamplification (PEP), degenerate oligonucleotide primed PCR (DOP-PCR), and multiple displacement amplification (MDA). Each of these methods has its own merits and limitations. FINDINGS: Effects of primer length and composition on amplification quality and quantity were explored in this study at two different temperatures (30 degrees C & 40 degrees C). New primer designs combined with elevated amplification temperature has significantly improved MDA as measured by amplification yield, genome coverage, and allele drop out (ADO) analysis. A remarkable finding was the comprehensive amplification, at 30 degrees C & 40 degrees C, of the human whole genome via the use of GGGCAGGA*N*G hotspot recombination consensus primer. Amplification was characterized by Affymetrix 10K SNP chip analysis. Finally, the use of new primer designs has suppressed the template-independent DNA amplification (TIDA) both at 30 degrees C and 40 degrees C. CONCLUSION: The use of new primers in this study combined with elevated incubation temperatures in MDA has remarkably improved the specificity, amplification yield, and suppressed TIDA. PMID- 19309529 TI - Solution structure of the parvulin-type PPIase domain of Staphylococcus aureus PrsA--implications for the catalytic mechanism of parvulins. AB - BACKGROUND: Staphylococcus aureus is a Gram-positive pathogenic bacterium causing many kinds of infections from mild respiratory tract infections to life threatening states as sepsis. Recent emergence of S. aureus strains resistant to numerous antibiotics has created a need for new antimicrobial agents and novel drug targets. S. aureus PrsA is a membrane associated extra-cytoplasmic lipoprotein which contains a parvulin-type peptidyl-prolyl cis-trans isomerase domain. PrsA is known to act as an essential folding factor for secreted proteins in Gram-positive bacteria and thus it is a potential target for antimicrobial drugs against S. aureus. RESULTS: We have solved a high-resolution solution structure of the parvulin-type peptidyl-prolyl cis-trans isomerase domain of S. aureus PrsA (PrsA-PPIase). The results of substrate peptide titrations pinpoint the active site and demonstrate the substrate preference of the enzyme. With detailed NMR spectroscopic investigation of the orientation and tautomeric state of the active site histidines we are able to give further insight into the structure of the catalytic site. NMR relaxation analysis gives information on the dynamic behaviour of PrsA-PPIase. CONCLUSION: Detailed structural description of the S. aureus PrsA-PPIase lays the foundation for structure-based design of enzyme inhibitors. The structure resembles hPin1-type parvulins both structurally and regarding substrate preference. Even though a wealth of structural data is available on parvulins, the catalytic mechanism has yet to be resolved. The structure of S. aureus PrsA-PPIase and our findings on the role of the conserved active site histidines help in designing further experiments to solve the detailed catalytic mechanism. PMID- 19309530 TI - Early prevention by L-Arginine attenuates coronary atherosclerosis in a model of hypercholesterolemic animals; no positive results for treatment. AB - BACKGROUND: Endothelial dysfunction (ED) is an independent predictor of cardiovascular events. ED is also a reversible disorder, and nitric oxide donors like L-arginine may promote this process. Despite the positive results from several studies, there are some studies that have shown that L-arginine administration did not improve endothelium-dependent dilation or the inflammatory state of patients. In this study the early and the late effects of L-arginine on coronary fatty streak formation and ED biomarkers were considered in hypercholesterolemic rabbits. METHODS: 36 white male rabbits randomly assigned in 3 groups. Rabbits were fed 1% high-cholesterol diet (LP group, n = 15), or high cholesterol diet with oral L-arginine (3% in drinking water) (EP group, n = 15) or standard diet (control group, n = 6) for 4 weeks (phase I). Afterward, all animals were fed normal diet for 4 weeks (phase II). In the second phase, L arginine was discontinued for EP group and was begun for LP group. The plasma levels of lipids, von Willebrand factor (vWF), and nitrite were compared before and after 4 and 8 weeks of experiment. Coronary fatty streak formation was measure after 4 and 8 weeks of experiment. RESULTS: The plasma levels of lipids were increased significantly in both groups of LP and EP after phase I. The hypercholesterolemia induced significant increased vWF release in LP group. The L arginine supplementation led to significant plasma nitrite increment in EP group. The vWF in LP group was higher than other groups (p < 0.05). By the end of phase II, despite of start of L-arginine supplementation for LP group and L-arginine discontinuation in EP group, there were significantly more fatty streaks lesions in LP group coronary arteries than EP group. Furthermore, L-arginine supplementation did not result in significant nitrite increment in LP group. CONCLUSION: Early prevention by L-arginine may be helpful to prevent the ED, but our study did not suggest the treatment. It seems reasonable to consider ED-aside from control the cardiovascular risk factors in primary prevention of atherosclerosis and its clinical outcomes before development of irreversible vascular damage. PMID- 19309531 TI - Reduction in corpora lutea number in obese melanocortin-4-receptor-deficient mice. AB - Obese melanocortin-4-receptor-deficient (MC4R-/-) male mice are reported to have erectile dysfunction, while homozygous MC4R-/- female mice are apparently fertile. A recently established obese mouse strain, carrying an inactivating mutation in the MC4R gene, revealed difficulties in breeding for the homozygous female mice. This prompted us to determine the presence of follicles and corpora lutea (CL) in ovaries of MC4R-/- mice aged 3-6 months in comparison to wild type (MC4R+/+) littermates. Serial sections of formaldehyde-fixed ovaries of mice with vaginal signs of estrus and metestrus were assessed for the number of healthy and regressing follicles and CL. The number of CL, as an estimate for the ovulation rate, decreased to zero during aging in MC4R-/- mice. The number of small- (diameter 100-200 micrometer) and large-sized follicles namely antral follicles (diameter >200 micrometer) were slightly increased in MC4R-/- compared to MC4R+/+ mice. Greater differences were found in very large to cystic follicles, which were more numerous in MC4R-/- mice. The number of regressing antral follicles was higher in the MC4R-/- group compared to the MC4R+/+ group. This was associated with a wide range in the number of collapsed zonae pellucidae as the last remnants of regressed follicles. A conspicuous hypertrophy of the interstitial cells was noted in 6-month-old MC4R-/- mice. In conclusion, cystic follicles and the reduction in CL number point to a decreased ovulation rate in obese MC4R-/- mice. PMID- 19309532 TI - Effects of a diet rich in sesame ( Sesamum indicum) pericarp on the expression of oestrogen receptor alpha and oestrogen receptor beta in rat prostate and uterus. AB - The expression of oestrogen receptors (ERalpha and ERbeta) in the prostate and uterus tissues of Wistar rats supplied for 8 weeks with a diet rich in sesame (Sesamum indicum) pericarp (30 %) was monitored. Eight male rats, aged 6 weeks, were divided into a control group fed on a normal diet, and an experimental one, provided with the normal diet enriched with 30 % sesame pericarp. A similar experiment was performed with female rats. At the end of the experiment, the prostate and uterus tissues were surgically removed and kept at - 80 degrees C for up to 2 months. Western blotting and quantitative real-time PCR (qRT-PCR) methods were used in order to investigate the levels of receptor proteins and mRNA. Significant increase in the expression of ERbeta in prostate and uterus was evident in both methods, while the magnitude of the observed alteration depended on the applied method. No statistically significant change was observed in the expression of ERalpha in uterus. In prostate, although the increase was more evident when investigated by means of qRT-PCR, the difference in expression of ERalpha was not statistically significant. In both tissues, a shift of the ratio of ERalpha:ERbeta in favour of ERbeta was evident, indicating, according to existing literature, a beneficial effect of the diet provided upon the health status of the organisms. It is suggested that this effect is attributed to the lignans present in the pericarp which exert phyto-oestrogenic activity. PMID- 19309533 TI - Image statistics at the point of gaze during human navigation. AB - Theories of efficient sensory processing have considered the regularities of image properties due to the structure of the environment in order to explain properties of neuronal representations of the visual world. The regularities imposed on the input to the visual system due to the regularities of the active selection process mediated by the voluntary movements of the eyes have been considered to a much lesser degree. This is surprising, given that the active nature of vision is well established. The present article investigates statistics of image features at the center of gaze of human subjects navigating through a virtual environment and avoiding and approaching different objects. The analysis shows that contrast can be significantly higher or lower at fixation location compared to random locations, depending on whether subjects avoid or approach targets. Similarly, significant differences in the distribution of responses of model simple and complex cells between horizontal and vertical orientations are found over timescales of tens of seconds. By clustering the model simple cell responses, it is established that gaze was directed toward three distinct features of intermediate complexity the vast majority of time. Thus, this study demonstrates and quantifies how the visuomotor tasks of approaching and avoiding objects during navigation determine feature statistics of the input to the visual system through the combined influence on body and eye movements. PMID- 19309535 TI - MAOA is associated with methylphenidate improvement of oppositional symptoms in boys with attention deficit hyperactivity disorder. AB - The monoamine oxidase A (MAOA) gene has been extensively related to aggressive, impulsive and violent behaviours. Previous studies have documented the improvement of oppositional symptoms in attention deficit hyperactivity disorder (ADHD) patients with methylphenidate (MPH). However, the effect of the MAOA gene in response to MPH has not been investigated. A sample of 85 boys from an ADHD outpatient service was genotyped for the MAOA-uVNTR polymorphism. The outcome measure was the parent-rated oppositional subscale of the Swanson, Nolan and Pelham Scale - version IV. The scale was applied by child psychiatrists blinded to genotype at baseline and in the first and third months of treatment. A significant interaction between the presence of MAOA high-activity genotype and treatment with MPH over time on oppositional scores was detected during the 3 months' treatment (n=85, F2,136=4.83, p=0.009). These results suggest an effect of the MAOA-uVNTR high-activity genotype on the improvement of oppositional symptoms with MPH treatment. PMID- 19309534 TI - Chronic vagus nerve stimulation induces neuronal plasticity in the rat hippocampus. AB - Vagus nerve stimulation (VNS) is used to treat pharmacotherapy-resistant epilepsy and depression. However, the mechanisms underlying the therapeutic efficacy of VNS remain unclear. We examined the effects of VNS on hippocampal neuronal plasticity and behaviour in rats. Cell proliferation in the hippocampus of rats subjected to acute (3 h) or chronic (1 month) VNS was examined by injection of bromodeoxyuridine (BrdU) and immunohistochemistry. Expression of doublecortin (DCX) and brain-derived neurotrophic factor (BDNF) was evaluated by immunofluorescence staining. The dendritic morphology of DCX+ neurons was measured by Sholl analysis. Our results show that acute VNS induced an increase in the number of BrdU+ cells in the dentate gyrus that was apparent 24 h and 3 wk after treatment. It also induced long-lasting increases in the amount of DCX immunoreactivity and in the number of DCX+ neurons. Neither the number of BrdU+ cells nor the amount of DCX immunoreactivity was increased 3 wk after the cessation of chronic VNS. Chronic VNS induced long-lasting increases in the amount of BDNF immunoreactivity and the number of BDNF+ cells as well as in the dendritic complexity of DCX+ neurons in the hippocampus. In contrast to chronic imipramine treatment, chronic VNS had no effect on the behaviour of rats in the forced swim or elevated plus-maze tests. Both chronic and acute VNS induced persistent changes in hippocampal neurons that may play a key role in the therapeutic efficacy of VNS. However, these changes were not associated with evident behavioural alterations characteristic of an antidepressant or anxiolytic action. PMID- 19309536 TI - Inhibitory processes in memory are impaired in schizophrenia: evidence from retrieval induced forgetting. AB - Schizophrenic patients are known to exhibit inhibitory impairments in response suppression and selective attention. However, the impairment of inhibitory control in memory retrieval has not clearly been documented. In two experiments, we investigate inhibition in memory retrieval by using the retrieval practice procedure. In Expt 1, a cued recall final test was used. Consistent with previous research, we found similar retrieval-induced forgetting (RIF) effects in schizophrenic patients and in controls. However, these effects could be the result of interference/blocking or the results of inhibition. In order to reduce the influence of blocking in Expt 2, we used a recognition test. We found that RIF was reduced in patients, compared to healthy controls. The elimination of RIF effect in patients, when the influence of blocking is reduced, indicates that inhibitory processes in memory are altered in schizophrenia. Result suggest that schizophrenic patients suffer from critical impairments in inhibitory processes involved in memory retrieval, similar to the inhibitory deficits found in other cognitive domains. PMID- 19309537 TI - Verbalization and problem solving: insight and spatial factors. AB - Two groups of participants attempted eight examples of each of four different problem types formed by combining insight versus non-insight and verbal versus spatial factors. The groups were given different verbalization instructions viz., Silent (N=40) or Direct Concurrent (N=40). There were significant differences between insight and non-insight tasks and between spatial and verbal tasks in terms of solution rates and latencies. Significant interactions between the verbal versus spatial factor and verbalization condition on solution rates and latencies reflected a greater (negative) effect of verbalizing on spatial as against verbal problems. However, no significant interactions of the insight versus non-insight factor with verbalization condition on solution rates or latencies were found. These results favoured the 'business as usual' view of insight problem solving as against the 'special process' view which predicted larger effects of verbalization for insight problems as against non-insight problems. PMID- 19309538 TI - Dietary supplement with fermented soybeans, natto, improved the neurobehavioral deficits after sciatic nerve injury in rats. AB - Clearance of fibrin and associated inflammatory cytokines by tissue-type plasminogen activator (t-PA) is related to improved regeneration in neurological disorder. The biological activity of fermented soybean (natto) is very similar to that of t-PA. We investigated the effect of the dietary supplement of natto on peripheral nerve regeneration. The peripheral nerve injury was produced by crushing the left sciatic nerve with a vessel clamp in Sprague-Dawley rats. The injured animals were fed orally either with saline or natto (16 mg/day) for seven consecutive days after injury. Increased functional outcome such as sciatic nerve functional index, angle of ankle, compound muscle action potential and conduction latency were observed in natto-treated group. Histological examination demonstrated that natto treatment improved injury-induced vacuole formation, S 100 and vessel immunoreactivities and axon loss. Oral intake of natto prolonged prothrombin time and reduced fibrinogen but did not change activated partial thromboplastin time and bleeding time. Furthermore, natto decreased injury induced fibrin deposition, indicating a tolerant fibrinolytic activity. The treatment of natto significantly improved injury-induced disruption of blood nerve barrier and loss of matrix component such as laminin and fibronectin. Sciatic nerve crush injury induced elevation of tumor necrosis factor alpha (TNF alpha) production and caused apoptosis. The increased production of TNF-alpha and apoptosis were attenuated by natto treatment. These findings indicate that oral intake of natto has the potential to augment regeneration in peripheral nerve injury, possibly mediated by the clearance of fibrin and decreased production of TNF-alpha. PMID- 19309539 TI - A modified rabbit model of stroke: evaluation using clinical MRI scanner. AB - OBJECTIVE: Occluding the middle cerebral artery of small animals with an intraluminal filament to build a stroke model has gained increasing acceptance. In light of the growing demand for magnetic resonance imaging (MRI) studies using the clinical MRI scanner, large animal models can be superior to small animal models. In this work, we developed a modified rabbit model of stroke, which was assessed using clinical MRI scanner and compared with a most commonly silicone coated filament model. METHODS: We presented a focal cerebral ischemia in rabbits. The key feature of this modified method is the use of a guide wire as a 'nylon suture'. At 3 days after ischemia, the percentage of brain infarct volume, neurobehavioral score, intracranial hemorrhagic incidence and dynamic changes of T(2) and apparent diffusion coefficient values were assessed respectively and compared between the focal cerebral models. RESULTS: Wire-induced models had more severe brain infarct size with less dispersion (32.7 +/- 6.5%, coefficient of variation=0.20) than that with filament models (25.4 +/- 8.9%, coefficient of variation=0.31; p<0.05). There were more significant MRI changes in the early stage, higher rate of technique success (wire, 20/20; filament, 17/20) and less intracranial hemorrhage (wire, 0/20; filament, 3/20) in wire-induced models than in filament-induced rabbits (p<0.05). CONCLUSION: Our data suggest that wire induced method can provide a useful tool for the earlier research of ischemia. PMID- 19309540 TI - A rat glioblastoma model with diffuse leptomeningeal gliomatosis induced by intracarotid injection of C6 glioma cells. AB - OBJECTIVE: A reproducible brain tumor model using experimental animals is required to study biological behavior and develop more potent antineoplastic drugs and effective therapeutic modalities. In this work, we attempted to establish diffuse leptomeningeal gliomatosis in the rat by intracarotid injection of C6 glioma cells. METHODS: Intracarotid injection of 1 x 10(7) C6 glioma cells in Wistar rats was performed to establish a primary diffuse leptomeningeal gliomatosis model. Ki-67 and matrix metalloproteinases (MMPs) immunohistochemistry staining were used to study the biological behavior of the developed tumor. Methodology, physical findings and histopathological features were also discussed. RESULTS: Leptomeningeal gliomas grew in all Wistar rats after the administration of 1 x 10(7) C6 glioma cells. Intracranial hypertension, weight loss and cachexia developed, and the median survival time was 18.0 +/- 2.9 days. The glioma mass distributed throughout the ventricles, the leptomeningeal regions in the brain and the brainstem, with typical pathological features of glioblastoma. The immunohistochemistry stainings showed high Ki-67 labeling index (42.1 +/- 10.3%), and concomitant overexpression of MMP-2 and MMP-9 suggested proliferation, invasion and angiogenesis potential. DISCUSSION: The advantage of the intracarotid injection route is the absence of an operative scar in the cranium. This established animal model is a novel model of primary diffuse leptomeningeal gliomatosis. This model probably can be used for pre-clinical testing in the progression of glioblastoma. PMID- 19309541 TI - Changes in neuropsychological functioning following temporal lobectomy in patients with temporal lobe epilepsy. AB - PURPOSE: This study was conducted to evaluate the changes in neuropsychological functioning in patients with temporal lobe epilepsy (TLE) after temporal lobe resection. METHODS: Fifty-four TLE patients were evaluated before and after surgery using comprehensive neuropsychological tests to assess general intelligence, executive functioning, language, verbal and visual memory, working memory, visuo-spatial ability, attention and motor function. RESULTS: The patients with left TLE showed no impairment of neuropsychological functioning after surgery, with the exception of auditory immediate memory. Furthermore, they showed significant improvement in performance IQ, executive function, working memory, visual memory, attention and psychomotor speed. The patients with right TLE did not show any significant impairment in post-operative neuropsychological functioning. They showed improvements in intellectual and executive functions, language, visual memory, visuo-spatial ability, attention and motor function post operatively. The patients with hippocampal sclerosis showed greater post operative improvements than the patients without hippocampal sclerosis regardless of the side. Patients with better pre-operative neuropsychological function had a higher chance of successfully discontinuing all seizure medications after surgery. DISCUSSION: The results of this study suggest that temporal lobectomy does not harm the neuropsychological functioning of patients with intractable TLE and that it improves cognitive functions of the contralateral hemisphere. PMID- 19309542 TI - Hypertonic saline more efficacious than mannitol in lethal intracranial hypertension model. AB - BACKGROUND: Medical management of brain edema and elevated intracranial pressure (ICP) is a crucial challenge in neurosurgical practice. Depending on the cause, the treatments for brain edema fall into three categories: stabilization of the blood-brain barrier, depletion of brain water and surgical decompression. Although mannitol is the mainstay of hyperosmolar therapy, hypertonic saline (HS) is emerging as an effective alternative to traditional osmotic agents. METHODS: Experimental elevated ICP (50 mmHg) was induced in rabbits using an intracranial balloon. The effects of mannitol and HS (10% NaCl) were compared in this specific physiopathological model. Twelve animals were divided into three groups (control, HS and mannitol) according to intravenous administration of 0.9% NaCl, 10% NaCl or 20% mannitol 5 minutes after the elevation of ICP. The doses of 10% NaCl and 20% mannitol were iso-osmolar. During 90 minutes, continuous recording of ICP, mean arterial pressure (MAP) and cerebral perfusion pressure (CPP) was realized. RESULTS: The control group had a median survival of only 53 minutes, significantly lower than the treated groups (p=0.0002). There was statistical difference between mannitol and HS; the 10% NaCl group had lower values of ICP (p=0.0116) and higher values of MAP (p<0.0001) and CPP (p<0.0001). CONCLUSION: The findings demonstrate higher efficacy of the 10% NaCl treatment in this comparison with 20% mannitol. Further efforts should be directed toward development of clinical studies using iso-osmotic doses of mannitol and HS in specific etiologies of intracranial hypertension. PMID- 19309543 TI - 5-lipoxygenase inhibitor zileuton attenuates ischemic brain damage: involvement of matrix metalloproteinase 9. AB - OBJECTIVES: Cerebral ischemia causes an increased expression and activation of 5 lipoxygenase (5-LOX) and matrix metalloproteinase 9 (MMP-9). Recent works demonstrated that the selective 5-LOX inhibitor zileuton down-regulates MMP-9 expression in vascular diseases and cancer. In the present work, we first studied the neuroprotective effect of zileuton on focal cerebral ischemia in rats and further investigated the effect of zileuton on the expression of 5-LOX and MMP-9 in ischemic brain. METHODS: Adult Sprague-Dawley rats underwent permanent middle cerebral artery occlusion (pMCAO). The rats then received treatment with zileuton 5, 10 or 50 mg/kg or vehicle. Cerebral water content and infarct volume were measured 24 hours after pMCAO. Expression of 5-LOX messenger RNA (mRNA) and MMP-9 mRNA were determined by reverse transcription polymerase chain reaction. The levels of the proform and the active form of MMP-9 were detected by gelatin zymography. RESULTS: Oral treatment of zileuton at 10 or 50 mg/kg significantly reduced cerebral water content and infarct volume. It also down-regulated the expression of 5-LOX mRNA and MMP-9 mRNA. Zileuton at 50 mg/kg could inhibit the active form of MMP-9. CONCLUSION: These results suggest that selective 5-LOX inhibitor zileuton attenuates ischemic brain damage in rats, which may be partly associated with MMP-9 inhibition. PMID- 19309544 TI - Morphometric study on dimensions of various parts of pons and comparison of data in accordance with age and sex of healthy people by MRI. AB - OBJECTIVES: The emergence of magnetic Resonance Imaging (MRI) technique has made it possible to answer many questions in the field of anatomy more precisely while observing different anatomic parts of living persons at different ages. This study was conducted to determine the dimensions of different parts of pons and also comparing the data in accordance with age and sex by MRI. METHODS: The present study carried out on 300 healthy individuals referred to imaging center at Imam Reza hospital in the city of Kermanshah, Iran. The precondition for people to be included in the study was the approval made by in-house physician based on person's state of health and the lack of any pathological lesion in brain on the basis of images obtained by MRI. Following MRI procedure, the dimensions of target parts in pons were calculated by the MRI-associated measuring system and recorded along with the age and sex of the patients. Measurements were recorded in mm and based on selecting the largest height, width and length of the organ during the examination of all axial, sagittal, and T(1) coronal profiles. Data analysis was performed using t-student test and Pearson's correlation. The significance level was set at P=0.05. RESULTS: The mean size of different parts of pons recorded in mm was as follows: The height of tegmentum of pons in men and women were 21.10+/-1.99 and 20.35+/-1.95, respectively. The height of ventricle of pons was 24.96+/-1.45 in men and 23.72+/-1.41 in women. The length of pons in men was 21.40+/-1.55 and in women 21.10+/-1.28. The length of ventricle in men and women were 16.85+/-1.21 and 16.77+/-1.36, respectively. The length of tegmentum of pons was 4.57+/-0.50 in men and 4.40+/-0.58 in women. Lastly, the transverse length of the ventricle of pons was 27.30+/-1.68 in men and 26.52+/-1.63 in women. DISCUSSION: The values obtained for the transverse length, length of tegmentum of ventricle, height of ventricle and height of tegmentum in men were larger than those found for women. There were significant relationships between the transverse length, length of tegmentum of pons, height of ventricle of pons and the age in men. There was no significant relationship between dimension of pons and age in women. PMID- 19309545 TI - Endogenous tissue plasminogen activator increases hemorrhagic transformation induced by heparin after ischemia reperfusion in rat brains. AB - OBJECTIVE: Tissue plasminogen activator (tPA) as a main thrombolytic drug for acute ischemic stroke remains complicated by risk of hemorrhagic transformation. However, whether endogenous tPA is also involved in hemorrhagic transformation is yet unclear. METHODS: We randomly assigned male Sprague-Dawley rats into three groups: the heparin group, the control group and the sham operated group. The ischemic rat models were induced by middle cerebral artery occlusion through intraluminal thread technique for 2 hours, followed by 24 hours of reperfusion. Heparin or saline was intermittent peritoneally injected after reperfusion. The extent of cerebral hemorrhage, the infarct volume, as well as the content and activity of endogenous tPA were evaluated. The matrix metalloproteinase 9 (MMP-9) expression and activity were also measured. RESULTS: All rats receiving heparin after reperfusion were subjected to hemorrhagic transformation. Hemorrhage volume in the heparin group was remarkably present. There was significant difference between the two groups (p<0.01). In the heparin group, the expressions of endogenous tPA and MMP-9 obviously increased, while their content and activity had significant differences compared with that of the control group (p<0.01). CONCLUSION: Endogenous tPA, through enhancement of MMP-9 expression and proteolytic activation, plays an important role in the pathogenesis of hemorrhagic transformation after cerebral reperfusion induced by heparin. PMID- 19309546 TI - Celiac disease in the 21st century: issues of under- and over-diagnosis. AB - Until the 1960s celiac disease (CD) or sprue was considered a pediatric disease that was rarely diagnosed in adulthood. Thanks to greater awareness of the disease and the availability of improved diagnostic tools (above all, sophisticated endoscopic techniques and the development of reliable serological markers), the prevalence of CD in Western countries has been increasing steadily, and it is now recognized as a common disorder, even in adults. However, many cases of this disease still go undiagnosed, especially among the elderly and in patients with atypical clinical presentations (which are by no means uncommon). On the other hand, the frequency of unfounded diagnoses of CD is also on the rise. This reflects a tendency toward exclusively symptomatic diagnosis as well as the growing use of invalidated tests for CD (e.g., the cytotoxic test, the sublingual or subcutaneous provocation/neutralization test, etc.). As a result, public healthcare spending is being increased in several countries (Italy included) by the growing number of prescriptions for gluten-free diets. This editorial discusses the problems of under- and over-diagnosis of CD and provides an algorithm for management of suspected cases designed to minimize both problems with particular importance to morphologic aspects of small bowel (also in electron microscopy), in basal conditions or in gluten-free diets. PMID- 19309547 TI - Chlamydia pneumoniae and atherosclerosis: current state and future prospectives. AB - Chlamydia pneumoniae, an intracellular bacterial pathogen, is known as a leading cause of human respiratory tract infections worldwide. Over the last decade, several reports in the literature have suggested that infection with C. pneumoniae may contribute to the pathogenesis of atherosclerosis. In order to play a causative role in chronic disease, C. pneumoniae would need to persist within infected tissue for extended periods of time, thereby stimulating a chronic inflammatory response. C. pneumoniae has been shown to disseminate systemically from the lungs through infected peripheral blood mononuclear cells and to localize in arteries where it may infect endothelial cells, vascular smooth muscle cells, monocytes/macrophages and promote inflammatory atherogenous process. The involvement of C. pneumoniae in atherosclerosis was investigated by seroepidemiological and pathological studies, in vivo and in vitro studies, and in clinical antibiotic treatment trials. This review will provide an update on the role of C. pneumoniae in atherosclerosis focusing on the recent insights and suggesting areas for future research. PMID- 19309548 TI - Autism and immunity: revisited study. AB - Autism spectrum disorder is of interest neurochemically because it represents a relatively homogeneous disorder with regard to disease development, abnormal cognitive development and intellectual development disturbance. A consistent finding in autistic children is a high number of mast cells and a high level of serotonin which is also found at elevated concentrations in the urine of autistic patients. In addition, a dysfunction of clinical conditions, such as gastrointestinal and immunological symptoms, is frequently noted in autistic children, however, IgE does not appear to be prevalent in these children but probably an increase of cytokines/chemokines produced by mast cells at an early age may play an important role. Therefore an immune hypothesis, involving also autoimmunity, is one possible pathogenetic mechanism in autism. In conclusion, mast cell activation could contribute to immune and neuroinflammatory abnormalities that are evident in patients with autism spectrum disorders. PMID- 19309549 TI - N-oleoyl-dopamine decreases muscle rigidity induced by reserpine in rats. AB - N-oleoyl-dopamine (OLDA), a product of condensation of oleic acid and dopamine (DA), is a bioactive compound that crosses the blood-brain barrier after systemic administration. The possibility arises that OLDA could have a potential role in treating DA-related disorders, such as Parkinsons disease (PD). In the present study we seek to determine whether OLDA would affect muscle tone and akinesia in two rat models of PD: the reserpine-evoked muscle rigidity and the reserpine- and haloperidol-induced catalepsy. We found that OLDA (20 mg/kg) significantly decreased muscle rigidity induced by reserpine (2.5 mg/kg), measured as an increased mechanical muscle resistance (MMG) in response to a passive extension and flexion of a rat hind limb at the ankle joint. Moreover, OLDA potently decreased the reserpine-enhanced tonic and reflex electromyographic (EMG) activities recorded before and during the movement, respectively. A lower dose of OLDA (10 mg/kg) failed to have appreciable effects. The reference compound L-DOPA (25 mg/kg) also attenuated the reserpine-increased MMG and EMG activities; the effects were, however, observed much later and were less prominent than those characteristic of OLDA. In contrast to the effects on muscle tone, OLDA (20 and 40 mg/kg) did not influence catalepsy induced by either reserpine (1.25 mg/kg) or haloperidol (0.5 mg/kg). In conclusion, the study demonstrates a novel biological action of N-oleoyl-dopamine consisting of lowering the reserpine-induced muscle rigidity. However, the lack of influence on akinesia suggests that the compound has myorelaxant rather than anti-Parkinsonian properties. PMID- 19309550 TI - The immunosuppressor st1959, a 3,5-diaryl-s-triazole derivative, inhibits T cell activation by reducing NFAT nuclear residency. AB - 3-(2-ethylphenyl)-5-(3-methoxyphenyl)-1H-1,2,4-triazole (ST1959) has shown therapeutic effects in several animal models of autoimmune diseases. In this study the effects of ST1959 were further investigated in a murine model of colitis. The evidence obtained indicates that the beneficial effects exerted by ST1959 rely upon a decreased local immunological response. The cellular effects of ST1959 were additionally investigated on human peripheral blood mononuclear cells and Jurkat T cells by measuring cytokine production, cell proliferation and activation of a set of transcription factors. ST1959 decreases human T cell proliferation and inhibits cytokine expression at the transcriptional level. Moreover, at doses inhibiting cytokine production, ST1959 blocks phorbol 12 myristate 13-acetate (PMA) and ionomycin-induced nuclear factor protein of activated T cell (NFAT1) activity, without impairing AP-1- and NF-kB-dependent transcription. Immunofluorescence data show that ST1959 inhibits the nuclear residency of NFAT1 in both Jurkat and human peripheral blood mononuclear cells activated with PMA/ionomycin. leptomycin B, an inhibitor of CRM1/exportin-1alpha dependent nuclear export, reverted the inhibitory effect of ST1959 on NFAT1 nuclear localization. This indicates that ST1959 may increase the nuclear export of NFAT1, downregulating NFAT1 activity via a mechanism different from that of cyclosporin A, since it does not affect NFAT phosporylation/dephosphorylation steps. These findings provide new insights into the molecular mechanisms underlying the immunomodulatory activity of ST1959. PMID- 19309551 TI - CXCR4+FOXP3+CD25+ lymphocytes accumulate in CXCL12-expressing malignant pleural mesothelioma. AB - CXCL12 is a chemokine that binds to a G-protein-coupled receptor (CXCR4). CXCL12 is expressed in various tumors and is considered as playing an important role in tumor growth and invasion. The aim of this study is to investigate the expression of CXCL12 in human malignant mesothelioma (MM), the chemotactic effect of CXCL12 derived from MM, and the expression of CXCR4 in MM tissues in relation to regulatory T cells. CXCL12 expression was examined by immunostaining of tissue specimens from malignant pleural mesothelioma (MPM) and malignant peritoneal mesothelioma (MPEM). The MM group comprised 6 patients (4 men/2 women, MPM=4, MPEM=2, aged 56.0 +/- 12.4 years) and the control (non-mesothelioma) group also had 6 patients (4 men/2 women aged 65.0 +/- 6.7 years). CXCL12 mRNA expression was also examined by RT-PCR in MPM cell lines (H28, H2052, and H2058), while CXCR4 mRNA expression was examined by in situ hybridization in MPM tissue. CXCL12 was expressed in the cytoplasm of MM cells from all patients, but was not expressed in the control group. H2052 and H2058 cells expressed CXCL12 mRNA, but H28 cells did not. CXCL12 in MM tissue homogenate supernatant had a chemotactic effect on CXCR4-expressing THP-1 cells. CXCR4 mRNA was expressed by a part of LCA+CD3+ Foxp3+CD25+ T cells that were located adjacent to the border of CXCL12 expressing epithelioid MPM. These findings suggest that CXCL12 contributed to tumor-related inflammation by inducing the accumulation of CXCR4-expressing cells with regulatory T cell markers around MM. PMID- 19309552 TI - Soluble interleukin-2 receptor as an indicator of immunological disturbance found in silicosis patients. AB - Silicosis patients (SILs) possess not only respiratory disorders but also alterations in autoimmunity. To determine an early indicator of immunological disturbance in SILs, the role of serum-soluble interleukin (IL)-2 receptor (sIL 2R) was analyzed. Of ten SILs, immunological clinical parameters such as immunoglobulin (Ig) G, complements, the titer of autoantibodies including anti nuclear antibodies (ANA), anti-Scl-70 antibody (Ab) and anti-centromere (CM) Ab, and experimental indicators such as serum-soluble Fas, serum IL-2, CD25+ cells in CD4+ or CD8+ fractions, and sIL-2R were divided from respiratory parameters such as percent vital capacity (%VC), percentage of forced expiratory volume in 1 second (FEV1.0%) and v25/Ht (liter/second/m(body height) by a correlation assay. Additionally, a stepwise regression test showed that sIL-2R was correlated with Ig G, ANA and anti-CM Ab. Furthermore, factor analysis revealed that sIL-2R contributed to the subpopulation of SILs with poorer immunological status in the absence of alterations in respiratory status. By defining healthy donors as 1, SILs as 2 and patients with systemic sclerosis as 3 for immunopathological progression status as metric variables, sIL2R and ANA showed a strong positive correlation. This suggests that sIL-2R is a good clinical indicator of immunological disturbance found in SILs without clinical manifestations of any disturbance in autoimmunity. Further analysis using a large-scale number of patients should be performed to confirm these findings. PMID- 19309553 TI - Immunomodulatory effects of 1,25-dihydroxyvitamin D3 on TH1/TH2 cytokines in inflammatory bowel disease: an in vitro study. AB - Crohn's disease (CD) is associated with a higher type-1-helper T cell (Th1) cytokine expression, whereas ulcerative colitis (UC) appears to express a modified Th2 response. In addition to its classic role in calcium homeostasis, calcitriol, the hormonal active form of vitamin D, exerts immunoregulatory effects such as modulation of Th1/Th2 cytokines. Therefore, calcitriol administration could modify immune dysfunction in CD and UC. Nine patients with UC (M/F: 5/4; mean age 47 years, remission(R)/active(A) disease: 7/2), 8 patients with CD (M/F: 2/6; mean age 36, R/A 5/3) and 6 healthy controls (HC) (M/F: 3/3, mean age 4) were enrolled. Peripheral blood was collected after a drug-washout of 15 days and peripheral blood mononuclear cells were stimulated with mitogens alone or in the presence of physiological concentrations of calcitriol (100 pg/ml). Type 1 (IL-2, TNF-alpha, IFN-gamma) and type 2 (IL-10) cytokine production was assayed on supernatants by ELISA. Compared to HC, TNF-alpha production was significantly higher both in UC (p=0.0002) and CD (p=0.0001) patients, at baseline and after incubation with calcitriol (UC p=0.0003, CD p=0.0009). The effects of calcitriol incubation were: 1) reduced IFN-gamma (p=0.024) and increased IL-10 (p=0.06) production in UC patients; 2) reduced TNF alpha production in CD (p=0.032); 3) no significant effects in HC. Calcitriol increased, albeit not significantly, IL-10 production in UC compared to CD patients (p=0.09). These results suggest an important modulatory role of vitamin D in the Th1/Th2 immune response. The observation that the effect of this modulation was different in CD compared to UC patients provides an interesting area of research into the pathogenesis and treatment of these inflammatory conditions. PMID- 19309554 TI - Selection and characterization of a novel agonistic human recombinant anti-TRAIL R2 minibody with anti-leukemic activity. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising natural anticancer therapeutic agent because through its death receptors, TRAIL R1 and TRAIL-R2, it induces apoptosis in many transformed tumor cells, but not in the majority of normal cells. Hence, agonistic compounds directed against TRAIL death receptors have the potential of being excellent cancer therapeutic agents, with minimal cytotoxicity in normal tissues. Here, we report the selection and characterization of a new single-chain fragment variable (scFv) to TRAIL-R2 receptor isolated from a human phage-display library, produced as minibody (MB), and characterized for the in vitro anti-leukemic tumoricidal activity. The anti TRAIL-R2 MB2.23 efficiently and specifically bound to membrane-associated TRAIL R2 on different leukemic cell lines and could act as a direct agonist in vitro, initiating apoptotic signaling as well as complement-dependent cytotoxicity and antibody-dependent cell cytotoxicity, providing a rationale for further investigations of MB2.23 in anticancer therapy. PMID- 19309555 TI - Andolast acts at different cellular levels to inhibit immunoglobulin E synthesis. AB - The anti-asthmatic agent andolast is thought to inhibit the release of allergic mediators, but its mechanism of action is not fully understood. We investigated whether the compound inhibits immunoglobulin E (IgE) synthesis and tested the hypothesis that andolast affects immunoglobulin class switching. Interleukin (IL) 4 and the interaction of CD40 expressed on B cells with its ligand on T cells are necessary for IgE synthesis. Thus, peripheral blood mononuclear cells (PBMCs) from 40 asthmatic, 16 non-asthmatic allergic, and 9 normal donors were stimulated with IL-4 and/or anti-CD40 antibody. T cells from 9 additional allergic donors were activated with anti-CD3/CD28 antibodies to express IL-4 mRNA. After incubation in the absence or presence of test compounds, immunoglobulin concentrations were measured by enzyme immunoassay, and mRNA levels were analyzed by RT-PCR. Andolast significantly inhibited IgE synthesis by stimulated PBMCs from both asthma patients and combined allergic/normal donors. In mechanistic studies, andolast was found to act at different cellular levels. Firstly, it reduced by about 45 percent (p<0.05) the levels of IL-4 mRNA in T cells stimulated with anti-CD3/CD28. Secondly, andolast reduced by about 36 percent (p<0.05) the expression of epsilon germline transcripts in PBMCs stimulated with IL-4/anti-CD40. Thirdly, the effect of andolast on immunoglobulin synthesis was selective in that the production of IgG4 antibodies was not significantly inhibited. Our findings, while supporting the evidence that andolast is effective for the treatment of asthma, provide new insights into its mechanism of action. PMID- 19309556 TI - Osteoblast apoptosis in periodontal disease: role of TNF-related apoptosis inducing ligand. AB - Periodontal disease (Pd) is characterized by an increased osteoclast resorption and a decreased osteoblast (OB) bone formation. OBs obtained from alveolar bone of Periodontitis patients (Pp) undergo apoptosis in the presence of TNF-related apoptosis-inducing ligand (TRAIL). We studied the intracellular apoptotic pathway induced by TRAIL; TRAIL death (DR4, DR5) and decoy (DcR1, DcR2) receptors expression in Periodontitis patients OBs (PpOBs), and we measured the concentration of TRAIL in the serum of Pp. We demonstrated that DNA fragmentation and activation of caspase-8 and caspase-3 in PpOBs, following TRAIL stimulation, occurred in shorter time; moreover, a higher amount of both caspases was activated in order to direct OBs. Down-regulation of DcR2 in PpOBs was demonstrated and high TRAIL levels were detected in the serum of Pp. In conclusion, our data suggest that PpOBs are more sensitive to TRAIL-induced apoptosis when compared to the control group. The down-regulation of DcR2 possibly leads to an imbalanced ratio between death and decoy receptors. Our findings highlight a role of TRAIL in the pathogenesis of Pd. PMID- 19309557 TI - Role of interleukin-15 receptor alpha polymorphisms in normal weight obese syndrome. AB - Previous published studies have identified a class of women, Normal Weight Obese women (NWO) with normal BMI and high fat content. An important role of Interleukin-15 (IL-15) has been documented in facilitating muscle proliferation and promoting fat depletion. Indeed the presence of three types of IL-15 receptor subunits in fat tissue suggests a direct effect on adipose tissue. We studied three single nucleotide polymorphisms (SNP) of IL-15R-alpha receptor gene and investigated their relationship with NWO phenotype. We considered two classes of women according to their BMI and percent fat mass (percent FAT), class 1: including 72 overweight-obese women (high BMI-high fat mass) and class 2: including 36 NWO (normal BMI, high fat mass). Three sites of Interleukin-15 receptor subunit a gene were examined, located respectively in exon4, exon5 intron-exon border and exon7. Genotyping of the identified polymorphisms was performed by restriction fragment length polymorphism. Haplotype frequency estimation was performed by using the Mendel-University of Chicago program. Odds ratio analyses were calculated by EPISTAT program. Highly significant differences were observed for exon 7- exon5 intron-exon border and exon 4-exon 7 haplotype distribution between class 1 and class 2 women. These results strongly support the hypothesis that genetic variability of the IL-15 receptor has an important role in body fat composition. Our data underscore previous findings that suggest a potential role of IL-15 cytokine in NWO syndrome. PMID- 19309558 TI - Allelic frequencies of 3' Ig heavy chain locus enhancer HS1,2-A associated with Ig levels in patients with schizophrenia. AB - Infectious and autoimmune pathogenic hypotheses of schizophrenia have been proposed, prompting searches for antibodies against viruses or brain structures, and for altered levels of immunoglobulins. Previous experiments have shown that allele frequencies of the Ig heavy chain 3' enhancer HS1,2*A are associated with several autoimmune diseases, suggesting a possible correlation between HS1,2 alleles and Ig production. To test this, we analyzed levels of serum Igs and HS1,2*A genotypes in two independent cohorts, one of 88 schizophrenic inpatients (24 women) and a second of 133 healthy subjects (59 women). Both groups were similar in the frequency of individuals with altered serum concentration of Ig classes and IgG subclasses (schizophrenia panel-80 percent; controls-68 percent). With the possible exception of a stabilizing effect of olanzapine, no psychopharmacological drug consumed during the month prior to serum sampling in the schizophrenia group significantly affected Ig levels. In both patient and control cohorts, an increased frequency of the HS1,2*2A allele corresponded to increased Ig plasma levels, while an increased frequency of the HS1,2*1A allele corresponded to decreased Ig plasma levels. EMSA analysis with nuclear extracts from human B cells showed that the transcription factor SP1 bound to the polymorphic region of both HS1,2*1A and HS1,2*2A while NF-kB bound only to the HS1,2*2A. We predict that differences in transcription factor binding sites in the two allelic variants of the 3' IgH enhancer HS1,2 may provide a mechanism by which differences in Ig expression are affected. PMID- 19309559 TI - In vitro evaluation of the efficacy of a new laser surface implant: cellular adhesion and alkaline phosphatase production tests. AB - Bone tissue reacts in different ways to implant surfaces with different patterns. The aim of this study is to understand which laser generated surface pattern produces the best cell adhesion in vitro, evaluating both the activity of the alkaline phosphatase and the cells adhering to titanium samples. Tests were carried out on titanium samples with sandblasted surfaces with laser-produced holes with diameters of 5, 10, and 20 microm, and on sandblasted titanium cylinders without holes as controls. The samples were inserted into culture medium containing SaOS-2 cells for 3, 7 and 10 days. The results showed that at days 3 and 7 the laser surfaces stimulated a higher production of alkaline phosphatase (ALP) compared to the data from the control group. At day 10 there were no significant differences between the test group and the control group. PMID- 19309560 TI - Plant-derived recombinant F1, V, and F1-V fusion antigens of Yersinia pestis activate human cells of the innate and adaptive immune system. AB - Plague is still endemic in different regions of the world. Current vaccines raise concern for their side effects and limited protection, highlighting the need for an efficacious and rapidly producible vaccine. F1 and V antigens of Yersinia pestis, and F1-V fusion protein produced in Nicotiana benthamiana administered to guinea pigs resulted in immunity and protection against an aerosol challenge of virulent Y. pestis. We examined the effects of plant-derived F1, V, and F1-V on human cells of the innate immunity. F1, V, and F1-V proteins engaged TLR2 signalling and activated IL-6 and CXCL-8 production by monocytes, without affecting the expression of TNF-alpha, IL-12, IL-10, IL-1beta, and CXCL10. Native F1 antigen and recombinant plant-derived F1 (rF1) and rF1-V all induced similar specific T-cell responses, as shown by their recognition by T-cells from subjects who recovered from Y. pestis infection. Native F1 and rF1 were equally well recognized by serum antibodies of Y. pestis-primed donors, whereas serological reactivity to rF1-V hybrid was lower, and that to rV was virtually absent. In conclusion, plant-derived F1, V, and F1-V antigens are weakly reactogenic for human monocytes and elicit cell-mediated and humoral responses similar to those raised by Y. pestis infection. PMID- 19309561 TI - In vitro detection of herpes simplex virus -1 and -2 infection with immunospecific GD3+-CL6-enhanced magnetic resonance imaging. AB - Herpes simplex virus infections are prevalent viral infections in humans. HSVs are also the most common cause of sporadic viral encephalitis (HSE). Magnetic resonance is the imaging method of choice for HSE because it provides the most sensitive method for detecting early lesions. The objective of this study is to set-up and in vitro test an experimental contrast agent specific for antigens present on HSV-infected cells, bound with a paramagnetic agent detectable by MR imaging. A selected anti-HSV HrFab was labelled with Alexa Fluor 488, 125I and Gd3+Cl6. In order to assess anti-HSV affinity and specificity, ELISA assays were performed. Vero cells infected with HSV strains were visualized by MRI using anti HSV HrFab/Gd3+Cl6 complex. Results of the ELISA tests demonstrated that the anti HSV HrFab labelled with Gd3+Cl6 showed similar affinity for the antigens while the 125I immunoconjugate showed reduced affinity. MRI confirmed high affinity and specificity of antibody for the detection of HSV infections. PMID- 19309562 TI - Necrotic cell death in human amniotic cells infected by Listeria monocytogenes. AB - Listeria monocytogenes can cause a placental-foetal infection that results in spontaneous abortion, premature labour, stillbirth, or neonatal sepsis and meningitis. Bacteria cross the maternofoetal barrier at the villous syncytiotrophoblast level and subsequently spread from the placenta to the fetus. L. monocytogenes is able to induce different kinds of death in a variety of cells. Murine hepatocytes, murine T and human B lymphocytes, and murine dendritic cells die by apoptosis, whereas bacterial infection of murine and human macrophages leads mainly to necrotic cell death. As we previously described the efficient infection and growth of L. monocytogenes in a human amniotic cell line, we investigated the fate of these cells in order to analyse the mode of cell death. Our results provide biochemical and morphological evidence of necrotic death induced by L. monocytogenes infection. PMID- 19309563 TI - Effect of SP3 silencing on cytokeratin expression pattern in HPV-positive cells. AB - In an attempt to understand the molecular factors underlying squamous cell carcinogenesis in HPV-infected oral and cervical tissues, we examined the Sp3 dependent cytokeratin expression in HPV-positive CaSki cells. Two sets of cytokeratins were examined: the simple epithelial CK 7, 8, 18, 19, and 20, which are generally expressed in simple epithelia and CK4, 10, 13, and 17, which are expressed in squamous epithelia. Two additional CK pairs, i.e. CK6/CK16 and CK4/CK13 were analyzed as controls of the proliferation/differentiation cell status, respectively. We report that Sp3 gene silencing specifically hits CK18 and CK19, which are markers of oral and cervical squamous tumors. These data may be of help in immunopathological definition of squamous carcinogenesis. PMID- 19309564 TI - Vaccination with Trichinella spirallis antigens increases CD8+ peripheral T cells and enhances the Th2 immune response in Leishmania infantum challenged mice. AB - In this study we investigate the effect of Trichinella spiralis vaccination on immune responses elicited in BALB/c mice challenged subcutaneously with 0.5 x 10 6 of Leishmania infantum promastigotes. Secretion of specific anti-L. infantum antibodies and changes in the number of CD4+, CD8+ T cell and CD19+ B cells in the peripheral blood were tested for the evaluation of immune responses. Immunization with low amounts of T. spiralis antigens induced depression in anti Leishmania specific antibodies of the IgG1 isotype, while no changes in the number of CD4+ and CD8+ T cell subpopulations or CD19+ B cells were observed. In contrast, high amounts of T. spiralis antigens induced an enhancement in anti Leishmania specific antibodies of total IgG and IgG1 isotype, increase of CD8+ T cell number and activation of CD19+ B cells, indicated by the co-expression of CD69 marker. Our results suggest that immunization with a certain dose of T. spiralis antigens in experimentally challenged mice with L. infantum leads to an increase of peripheral CD8+ T cells which are responsible for the control of L. infantum infection, although a simultaneous enhancement in Th2-type of immune response is also observed. PMID- 19309565 TI - Exploiting immunotherapy in Mycobacterium tuberculosis-infected mice: sphingosine 1-phosphate treatment results in a protective or detrimental effect depending on the stage of infection. AB - Sphingosine 1-phosphate (S1P) is a natural lysophospholipid able to enhance antimycobacterial innate immune response. In the present study, we address the possible therapeutic role of S1P administered during primary or acute infection in mice aerogenically infected with Mycobacterium tuberculosis (MTB). Results show that the administration of S1P during primary infection significantly reduces the presence of MTB-infected cells within pulmonary granulomas and mycobacterial burden in the lung and in the spleen. However, if S1P treatment was started during acute infection, a detrimental effect was observed in terms of pulmonary histopathology and mycobacterial burden in the lung and in the spleen. Taken together, these results show that S1P can exert a therapeutic effect as a treatment of primary infection only. PMID- 19309566 TI - Expression of internalin A and biofilm formation among Listeria monocytogenes clinical isolates. AB - Internalin A (InlA), a cell wall-bound protein of Listeria monocytogenes, is among the major components involved in the adhesion to and invasion of host cells expressing specific forms of E-cadherin. Some L. monocytogenes strains secrete truncated non-functional forms of InlA. The purpose of this study is to compare the biofilm-forming abilities of L. monocytogenes strains from clinical sources expressing InlA proteins in the different forms. A total of 70 L. monocytogenes strains were examined using SDS-PAGE, Western blot, DNA sequencing, and microtitre plate biofilm formation assays. We found that 8 of the 70 strains expressed truncated InlA, and that this group of strains exhibited significantly enhanced biofilm-forming ability compared to the group expressing full-length InlA. Further experiments showed that: (i) L. monocytogenes biofilms were detached by treatment with protease K; (ii) protein fragments resulting from proteolysis, rather than intact proteins, are responsible for biofilm enhancement, because biofilm formation was impaired by the protease inhibitor alpha2-macroglobulin; (iii) truncated and/or proteolytically cleaved InlA are likely involved in the biofilm enhancement, based on the effects that anti-InlA monoclonal antibodies produced on the biofilm formation of L. monocytogenes strains expressing either truncated or full-length InlA. These data provide a basis for further investigation of the molecular structure and composition of L. monocytogenes biofilms. PMID- 19309567 TI - Analysis of memory and effector CD8+ T cell subsets in chronic graft-versus-host disease. AB - In humans, the selective depletion of CD8+ cells may prevent GVHD after allogeneic transplantation. These cells can infiltrate and damage target tissues. It is of interest to investigate the phenotypical characteristics and cytotoxic properties of the different CD8+ subsets in cGVHD patients. In a preliminary study we found that patients with cGVHD had a markedly elevated percentage of peripheral blood CCR7-/CD45RA+ cells compared to patients without cGVHD; conversely, the CCR7+/CD45RA+ subsets of CD8+ cells was significantly decreased. In this study, we report in depth on the phenotype of effector T cell subsets in cGVHD patients, as well as their proliferative capability, cytotoxic properties and cellular turnover. We confirm a predominance of effector T cell subsets in cGVHD patients and show that a large fraction of these cells down-regulate CCR7 and re-express CD45RA, thus approaching end-stage differentiation. Moreover CD8+ cells of cGVHD patients have low CD8 coreceptor expression, reduced proliferative potential and a high content of perforin and granzyme A. They also have a lower cell turnover and have more propensity to apoptosis, as demonstrated by BrdU incorporation. Taken together, our findings indicate a perturbation of the balance between naive/memory and effector/CD45RA+ CD8+ T cells, and suggest an involvement of the latter compartment characterized by a high content of cytotoxic equipment, in the pathogenesis of cGVHD. PMID- 19309568 TI - Duodenal intraepithelial lymphocytes of children with cow milk allergy preferentially bind the glycan-binding protein galectin-3. AB - A breakdown in intestinal homeostasis results in inflammatory bowel diseases including coeliac disease and allergy. Galectins, evolutionarily conserved beta galactoside-binding proteins, can modulate immune-epithelial cell interactions by influencing immune cell fate and cytokine secretion. In this study we investigated the glycosylation signature, as well as the regulated expression of galectin-1 and -3 in human duodenal samples of allergic and non-allergic children. Whereas galectin-1 was predominantly localized in the epithelial compartment (epithelial cells and intraepithelial lymphocytes) and the underlying lamina propria (T cells, macrophages and plasma cells), galectin-3 was mainly expressed by crypt epithelial cells and macrophages in the lamina propria. Remarkably, expression of these galectins was not significantly altered in allergic versus non-allergic patients. Investigation of the glycophenotype of the duodenal inflammatory microenvironment revealed substantial alpha2-6-linked sialic acid bound to galactose in lamina propria plasma cells, macrophages and intraepithelial lymphocytes and significant levels of asialo core 1 O-glycans in CD68+ macrophages and enterocytes. Galectin-1 preferentially bound to neutrophils, plasma cells and enterocytes, while galectin-3 binding sites were mainly distributed on macrophages and intraepithelial lymphocytes. Notably, galectin-3, but not galectin-1 binding, was substantially increased in intraepithelial gut lymphocytes of allergic patients compared to non-allergic subjects, suggesting a potential role of galectin-3-glycan interactions in shaping epithelial-immune cell connections during allergic inflammatory processes. PMID- 19309569 TI - Influence and variation of the body mass index in patients treated with etanercept for plaque-type psoriasis. AB - A relationship between psoriasis, pro-inflammatory cytokines and obesity has been demonstrated. Tumour necrosis factor-alpha (TNF-alpha), that is involved in the pathogenesis of psoriasis, is commonly over-expressed in obese subjects, and seems to be derived from inflammatory cells and adipocytes. The primary aim of this study is to investigate whether the Body Mass Index (BMI) of patients influences the clinical response to etanercept, a competitive inhibitor of TNF alpha approved for the treatment of moderate-to-severe plaque-type psoriasis. The secondary aim is to evaluate whether the TNF-alpha inhibition influences the weight and BMI profile of patients. One hundred patients received 50 mg etanercept twice weekly for 12 weeks, followed by 25 mg. At weeks-12 and 24, treatment efficacy and tolerability were evaluated, as well as body weight and BMI. BMI values did not correlate with etanercept efficacy. Mean PASI score variation did not show significant differences among the BMI groups. A statistically significant weight gain and BMI variation were observed in a consistent rate of patients. Patient BMI does not influence psoriasis efficacy parameters. Although the role of anti TNF-alpha molecules on weight regulation need to be confirmed, our study shows that etanercept treatment may induce weight gain and a BMI increase. PMID- 19309570 TI - A multicenter open-label experience on the response of psoriasis to Adalimumab and effect of dose escalation in non-responders: the Aphrodite project. AB - There is much evidence to show the efficacy of adalimumab, a human monoclonal antibody targeting tumour necrosis factor-alpha, in the treatment of plaque psoriasis. In this open-label experience, 147 high-need patients suffering from plaque psoriasis, with a mean Psoriasis Area and Severity Index (PASI) of 18.8, and concomitant psoriatic arthritis (PsA) received subcutaneous injections of 40 mg of adalimumab every other week (EOW). This was actually the dosage regimen recommended for PsA, as the drug had not then been approved for psoriasis at the time of the patients enrolment. At week 12, an improvement of at least 50 percent of the PASI (PASI-50) was observed in 111 (77 percent) patients. Continuation of treatment in responders with adalimumab 40 mg EOW led to a sustained response, with the PASI-50 achieved by 97 percent of patients in the as-treated analysis at week 24 (PASI-75 in 82 percent and PASI-90 in 45 percent out of 109 patients who received EOW injections up to week 24). Thirty subjects who failed to attain the PASI-50 response at week 12 were treated with adalimumab 40 mg every week for a further 12 weeks. At week 24, 80 percent of these patients obtained a PASI-50 response after dose escalation. Tolerability was good in the majority of patients. Only two patients discontinued treatment because of an adverse event (repeated flu-like episodes and a pleuropericarditis of unknown origin, respectively). PMID- 19309571 TI - Efficacy of cyclosporine in the treatment of a case of infliximab-induced erythrodermic psoriasis. AB - A clinical case is described of infliximab-induced erythrodermic psoriasis resistant to conventional topical therapy and high-dose corticosteroids. Cyclosporine therapy for some months resolved the severe psoriasis picture. Pathogenetic mechanisms are considered through which anti-TNF agents could induce psoriasis. An activation of T lymphocytes with cutaneous overexpression of a CXCR3 subset and, mainly, an increase in IFN-alpha due to the blockage of TNF alpha could be the causes for this paradoxical adverse event of biological agents. Cyclosporine can work in psoriasis induced by biological agents thanks to its peculiar suppressive activity on T lymphocytes and the concomitant specific action on keratocytes and angiogenesis. PMID- 19309572 TI - The erythromycin-resistance in S. pyogenes does not limit the human polymorphonuclear cell antimicrobial activity. AB - In order to highlight the potential erythromycin immunomodulatory properties related to different antibiotic resistance patterns in Streptococcus spp., we evaluated the influence of the macrolide on the PMNs primary functions against erythromycin-susceptible (Ery-S) and erythromycin-resistant (Ery-R) S. pyogenes strains. A total of 438 S. pyogenes were isolated over the period 2005-2007. On the basis of the triple disk testing, 345 out of 438 S. pyogenes isolates were Ery-S and 93 were Ery-R; among the resistant strains, 65 displayed the cMLSB phenotype, 23 had the M phenotype and 5 had iMLSB phenotype. Concerning antibacterial activity of PMNs, our results showed that erythromycin did not modify bacterial uptake, but significantly increased the phagocyte intracellular killing, compared with controls, for both Ery-S and Ery-R strains. Consequently, this report underlines that in immunocompetent hosts the dichotomy between the in vitro resistance and clinical trial data for antimicrobial agents should be thoroughly re-evaluated. PMID- 19309573 TI - Successful cyclosporine treatment in a case of amicrobial pustulosis associated with immunological abnormalities. AB - Amicrobial pustulosis associated with autoimmune diseases (APAD) is a clinical entity which was described only recently and few cases are reported in the literature. This condition is characterized by recurrent acute onset with pustular lesions predominantly involving skin folds, genitals, scalp and external auditory canals of young women. The etiopathogenesis of APAD is unknown and the most effective therapeutic treatment seems to be systemic corticosteroids. We describe the case of a 16-year old female patient suffering from APAD successfully treated with cyclosporine A. PMID- 19309574 TI - Specific immunotherapy for allergic rhinitis in Italy: the patients points of view. AB - Specific immunotherapy (SIT) is the unique causal treatment for allergy, but its use is quite limited. A perspective, cross-sectional telephone interview survey was carried out in Italy to evaluate the characteristics of 500 patients with allergic rhinitis (250 of whom treated with SIT). Relevant differences were found concerning therapeutic management of allergic rhinitis, mainly regarding the use of drugs and co-morbidities. The allergist is the most important consultant who prescribes SIT. This study therefore provides evidence that the course of allergic rhinitis may depend on the therapy prescribed by and the level of allergy awareness of the physician. PMID- 19309576 TI - [To strengthen the intervention of occupational stress for the working population's health and wellbeing]. PMID- 19309575 TI - Specific immunotherapy for allergic rhinitis in Italy: the doctors points of view. AB - Specific immunotherapy (SIT) is the unique causal treatment for allergy, but its prescription is quite restricted. A perspective and cross-sectional survey based on telephone interviews was carried out in Italy to evaluate the profile of doctors prescribing SIT for allergic rhinitis. A total of 540 doctors were interviewed, 200 of whom are GPs, 60 allergists, 60 ENT specialists, 100 familial paediatricians, 60 hospital paediatricians and 60 pulmonologists. Significant differences concern diagnostic and therapeutic management of allergic rhinitis, mainly regarding SIT prescription. The allergist is the most important consultant who prescribes SIT, as opposed to the paediatrician. This study therefore provides the evidence that doctors behaviour towards SIT depends on the type of graduate studies. PMID- 19309577 TI - [Influence of affectivity trait on relationships between occupational stress and subjective physical health and job satisfaction]. AB - OBJECTIVE: To explore the influence of affectivity trait on relationships between occupational stress and subjective physical health and job satisfaction. METHODS: Self-reports for psychosocial work conditions and health and well-being in a sample of 878 workers from a thermal power plant in China were conducted using the job demand-control model, the effort-reward imbalance model, job satisfaction, depression symptom, and physical health complaints questionnaires. Logistic regression analysis was used by controlling for age, sex, and educational level. RESULTS: Negative and positive affectivity were correlated with occupational stressors or strains (P<0.01). In the most equation of job dissatisfaction, psychosomatic complaints and depressive symptoms as dependent variables respectively, odds ratios decreased greatly when negative affectivity or positive affectivity was controlled; Odds ratios decreased greater when negative affectivity and positive affectivity were controlled simultaneously. Furthermore in some equation no variable entered. CONCLUSION: Affectivity trait has effect on relationships between occupational stress and subjective physical health and job satisfaction. According to the outcome category of study, the possible confounding role of affectivity should be controlled in the study of the relationships between occupational stress and health self-reports. PMID- 19309578 TI - [Relationship of occupational stress and negative life events with low birth weight]. AB - OBJECTIVE: To explore the influence of occupational stress and negative life events on low birth weight (LBW). METHODS: 1:1 matched case-control study was employed, in which 438 singleton LBW infants with birth weight less than 2500 g (their pregnancy term being 28 to 42 weeks) served as case group while 438 with singleton term normal birth weight served as control group matched by sex, delivery time and hospital. All of their mothers were inquired by well trained investigators about their socio-demographic characteristics, occupational stress, and negative life events occurring in different pregnancy term. After controlling for mother's age, occupation, education level and family income, conditional logistic regression was employed to asses the influence of occupational stress and negative life events on LBW. RESULTS: Compared with those with low technical skill utilization and low job decision, mothers with high technical skill utilization (OR=0.62; 95% CI=0.43 approximately 0.91) and high job decision (OR=0.67; 95% CI=0.46 approximately 0.97) significantly decreased the risk of laboring LBW. Compared with those not exposed to negative life events, mothers with negative life event score being=3 in the middle three months of pregnancy (OR=18.85; 95% CI=1.58 approximately 225.02), with negative life event score being 1 in the later three months of pregnancy (OR=2.67; 95% CI=1.14 approximately 6.28), with negative life event score being 2 (OR=2.80; 95% CI=1.04 approximately 7.52) and=3 in the whole time of pregnancy (OR=2.94; 95% CI=1.22 approximately 7.09) were the risk factors of LBW. CONCLUSION: Negative life events might affect LBW and negative life events occurring in the different term of pregnancy impact LBW differently. PMID- 19309579 TI - [Analysis of occupational stress in workers of a steel plant]. AB - OBJECTIVE: To investigate level and its influential factors of occupational stressors and strains in workers of a steel plant and explore the feasibility of occupational stress and mental health as indicators of effect evaluation of health promotion at workplace. METHODS: 1039 workers were investigated by using cross-sectional method. The questionnaires included job stressors, skill discretion, job control latitude, mental health and sleep disorders. RESULTS: 54.5% of respondents reported "job doesn't allow mistakes" as stressor, while 47.6% of respondents reported "too job responsibility" as stressor. The scores of job stressors were significantly higher than female workers (P<0.01). Workers with high education level scored more skill discretion, job control latitude and lower mental health than ones with low educational level (P<0.05 or P<0.01). Elder workers had more skill discretion and job control latitude scores than younger. There were significant differences in scores of skill discretion among different length of service groups (P<0.05). There were also significant differences in scores of skill discretion among different job title groups (P<0.01). Occupational stressors were correlated significantly positively with sleep disorder (r=0.37, P<0.01), but job control latitude negatively to mental health (r=-0.19, P<0.01). CONCLUSION: The workers in the steel plant report occupational stressors, which is related to poor self-reported health symptoms. The occupational stressors and mental health should be considered as one of effect evaluation indicators of health promotion at workplace. PMID- 19309580 TI - [Coping resources of nurses' occupational stress]. AB - OBJECTIVE: To study the effects of coping resources on nurses' strain. METHODS: Coping resources (recreation, self-care, social support, and rational coping) and strain (vocational strain, psychological strain, interpersonal strain, and physical strain) were measured on 387 nurses with personal resources questionnaire (PRQ) and personal strain questionnaire (PSQ). RESULTS: The higher the nurses' coping resources were, the lower their strain was (P<0.01). Nurses' coping resources had a negative correlation with their strain and had the closest relation with their psychological strain (P<0.01). Among coping resources, social support was closest related with nurses' strain. In the items of social support, those related to feeling support had more remission on strain. In the items of rational coping, those related to keeping good mind had more remission on strain, while doing more than one job at one time could raise nurses' strain (P<0.05). In the items of self-care, those related to good healthy habit, being immersed in contemplation and sufficient sleeping had the most effects on the remission of strain. In the items of recreation, those related to enough rest and self determination had more effects on the remission of strain, while doing other things in most of leisure time had a positive correlation with strain (P<0.05). CONCLUSION: Enhancing coping resources, especially feeling support, keeping good mind, sufficient sleeping and rest, are the important measures for the remission of nurses' strain. PMID- 19309581 TI - [Investigation on job stress of pediatricians and nurses working in pediatric department]. AB - OBJECTIVE: To investigate the occupational stressors and modifiers of pediatricians and nurses in order to find the measurements for control of the job stress. METHODS: 427 pediatricians and nurses working in five hospitals of a city served as subjects. Of them, the staff in section of pharmacy and toll offices in each hospital mentioned above served as control group. The General Job Stress Questionnaire was used to investigate the job stress by self-assessment. RESULTS: The scores of job demand, job risk, drug using, daily job stress, positive feelings, patient A behavior, physical environment and feeling balance in pediatricians and nurses were higher than those of control group, but the scores of job-person conflict, environmental control, technology utility, mental health, responsibility on things were lower than those of control group (P<0.05). The points of job future, job locus of control, self-esteem, job satisfaction, job load variance, depression in nurses were higher than those of pediatricians, and non-work activities, job risk and daily life stress were lower than those of doctors (P<0.05). The main affecting factors on job strain of pediatric staff included job monotony, higher job demand, more non-work job, lower job control, more job risk, job future ambiguous, poorer social support, lower job locus control and lower self-esteem. CONCLUSION: The stress degree of pediatric staff is higher than that of controls. The pediatricians have more job stress than that of nurses. The main stressors of pediatric staff are job monotony, higher job demand, more non-worker activity, lower job control, higher job risk and ambiguous job future. The main modifiers are good social support, external job locus of control and higher self-esteem. PMID- 19309582 TI - [Influence of electromagnetic radiation on raf kinase inhibitor protein and its related proteins of hippocampus]. AB - OBJECTIVE: To study the development of changes for Raf kinase inhibitor protein (RKIP) and its mRNA in rats hippocampus after electromagnetic radiation. METHODS: Rats were exposed to X-band high power microwave (X-HPM), S-band high power microwave (S-HPM) and electromagnetic pulse (EMP) radiation source respectively. The animal model of electromagnetic radiation was established. Western blot was used to detect the expression of RKIP, and RT-PCR was applied to detect the expression of RKIP mRNA. The interaction of RKIP and Raf-1 was measured with co immunoprecipitation method, and the expression of cerebral choline acetyltransferase (CHAT) was measured by immunohistochemistry. RESULTS: The expression of RKIP significantly down-regulated at 6 h after radiation, and recovered at 1 d in group EMP, but the down-regulation continued during 1 approximately 7 d after radiation in the two microwave groups. The expression of RKIP mRNA changed wavily during 6 h approximately 7 d after radiation, which showed down-regulation at 6 h, and up-regulation at 3 d. The interaction of RKIP and Raf-1 decreased during 6 h approximately 7 d after radiation, most significantly at 7 d, and the two microwave groups were more significant. The expression of CHAT decreased continuously during 6 h approximately 7 d after radiation, and generally recovered on 14 d. CONCLUSION: The down-regulation of RKIP and its related proteins of hippocampus is induced by electromagnetic radiation. PMID- 19309583 TI - [Construction and identification of antisense c-Jun N-terminal kinase 1 eukaryotic fluorescent expressing plasmids and JNK1-/- human embryo lung fibroblasts cell line]. AB - OBJECTIVE: To construct antisense c-Jun N-terminal kinase 1 (JNK1) eukaryotic fluorescent expressing vector and JNK1-/- human embryo lung fibroblasts cell line. METHODS: Trizol reagent was used to extract total RNA in HELF. The proper primers of JNK1 were chosen and synthesized. RT-PCR and gene recombinant techniques were used to construct the fragment of JNK1. After purification, the PCR products were cut, and JNK1 were inserted reversely into eukaryotic fluorescent expressing vector pEGFP-C1. Enzyme-cutting and DNA auto-sequencing were used to prove the successful construction of JNK1 eukaryotic expressing vector. Then plasmids were extracted and transfected into HELF cells and screen by G418 24 h later. Monoclone was chosen and cultured. Fluorescent imaging and Western blot were used to identify the JNK1-/- HELF cell line. RESULTS: Sequence analysis of pEGFP-C1-as JNK1 plasmids was same as expected. The expression level of JNK1 was inhibited markedly. CONCLUSION: Construction of antisense JNK1 eukaryotic fluorescent expressing vectors and JNK1-/- HELF cell line is successful. PMID- 19309584 TI - [Effect of Hg2+ on voltage-dependent calcium channels and intracellular free calcium in trigeminal ganglion neurons of rats]. AB - OBJECTIVE: To investigate the effects of Hg2+ on voltage-dependent calcium channels and intracellular free calcium in trigeminal ganglion neurons of rats and explore the toxicity mechanism of Hg2+ on these neurons. METHODS: Whole cell patch-clamp technique was used to determine ICa of voltage-dependent calcium channels in trigeminal ganglion neurons of rats. Intracellular free calcium was measured to explore [Ca2+]i dynamic changes from a single cell level by laser scanning confocal microscopy and fluorescence probe techniques. RESULTS: 0.01, 0.10, 1.00 and 10.00 micromol/L Hg2+ could reduce voltage-dependent calcium channel currents ICa by (1.80+/-0.32)%, (23.04+/-9.46)%, (58.20+/-7.90)% and (82.00+/-5.77)% in trigeminal ganglion neurons. The inhibiting effects reached their maximum in 5 minutes and could not be reversed significantly during wash with Hg2+-free solution. Also, 0.01, 0.10 and 1.00 micromol/L Hg2+ increased intracellular free calcium concentrations by (2.50+/-0.83)%, (82.81+/-35.36)% and (222.70+/-62.48)% in trigeminal ganglion neurons. Pre-administrated trigeminal ganglion neurons with nifedipine for 10 minutes could decrease the effects and delay the effecting time. CONCLUSION: The inhibition of Hg2+ on the voltage dependent calcium channel currents ICa depends on voltage-dependent calcium channels. And the increase of intracellular free calcium concentration in trigeminal ganglion neurons induced by Hg2+ is related to the release of intracellular stored calcium. However, the relationship between them needs further investigation. PMID- 19309585 TI - [Influence of benzo[a]pyrene on learning and memory and content of amino acid neurotransmitters in hippocampus of rats]. AB - OBJECTIVE: To study the effects of benzo[a]pyrene (B[a]P) on capability of learning and memory and the content of amino acid neurotransmitters in hippocampus of rats. METHODS: Thirty-two healthy, male SD rats were randomly divided into 4 groups according to their weights after intubated into ventricles: the solvent control group, 2.5, 5.0 and 10.0 mmol/L groups. 10 microl of B[a]P olive oil solutions, of different concentrations 2.5, 5.0 and 10.0 mmol/L, were injected into rats' lateral ventricles, respectively. Rats in the solvent control group were injected into the same volume of olive oil as that in B[a]P group. Rats' capability of learning and memory was tested by Morris water maze. The content of amino acid neurotransmitters in rats' hippocampus were determined by high performance liquid chromatogram with a fluorescence detector. RESULTS: Compared with the controls, the performances of learning and memory of rats decreased significantly in B[a]P treated groups (P<0.01). Levels of glutamate (Glu) were lower significantly in treated groups than that in controls (P<0.01). No significant differences were found in contents of aspartic acid (Asp), glycine (Gly) and aminobutyric acid (GABA) among the four groups. CONCLUSION: B[a]P can damage rats' spatial learning and memory, and which could be related to decreased contents of excitatory amino acids in hippocampus. PMID- 19309586 TI - [Clinical study on treatment of acute paraquat poisoning]. AB - OBJECTIVE: To investigate the clinical therapeutic effect of methylprednisolone combined with cyclophosphamide and Etanercept method on acute paraquat poisoning. METHODS: 136 patients with acute paraquat poisoning were divided into the normal therapy group and the intensive therapy group randomly. Methylprednisolone, cyclophosphamide and Etanercept were used in the intensive therapy group. Methylprednisolone 500 mg was given intravenously per day for continuous three days followed by 200 mg intravenous per day. Then methylprednisolone was decreased gradually 14 d or 21 d later according to the patient's condition. Cyclophosphamide 600 mg was given intravenously twice weekly for 2 weeks and Etanercept 25 mg was given hypodermic injection twice weekly for 3 weeks. Curative effect evaluation was done at 7, 14, 21 d and 12 weeks after therapy. RESULTS: The survival rate of the intensive therapy group was obviously higher than that of the normal therapy group (P<0.01) on 7, 4, 21 d and 12 weeks. The cure rate of the intensive group were 94.6% (intake dose<50 ml 20% paraquat solution), 75.0% (intake dose 50 approximately 100 ml 20% paraquat solution), 12.5% (intake dose>100 ml 20% paraquat solution) respectively, while the cure rate of the normal group were 16.7% (intake dose<50 ml 20% paraquat solution), 8.3% (intake dose 50 approximately 100 ml 20% paraquat solution), 0% (intake dose>100 ml 20% paraquat solution) respectively. The total cure rate of the intensive therapy group (78.3%) 12 weeks later was higher than that of the normal group (11.9%). CONCLUSION: Methylprednisolone combined with cyclophosphamide and Etanercept intensive therapy has the curative effect on acute paraquat poisoning. PMID- 19309587 TI - [A case of death induced by acute carbon dioxide poisoning]. PMID- 19309588 TI - [Fatigue symptoms of medical staff and influential factors]. PMID- 19309589 TI - [Survey on status quo of occupational stress in workers exposed to radiation]. PMID- 19309590 TI - [Survey on occupational disease hazards for farmer-workers in Guangzhou]. PMID- 19309591 TI - [Serum levels and clinical significance of IL in patients with delayed encephalopathy after acute carbon monoxide poisoning]. PMID- 19309592 TI - [Progress in study and application of cytokinesis-block micronucleus cytome assay]. PMID- 19309593 TI - [Progress in experimental study on mechanism of effect of oxidative stress on noise-induced hearing loss]. PMID- 19309594 TI - [Zeeman graphite furnace atomic absorption spectrophotometric determination of urine vanadium]. PMID- 19309595 TI - [Two cases of occupational chronic high altitude disease]. PMID- 19309596 TI - [A case of severe ammonia poisoning]. PMID- 19309597 TI - [Six cases of industrial fluorosis]. PMID- 19309598 TI - [Investigation on accidents of hydrogen arsenide poisoning]. PMID- 19309599 TI - [Risk factor analysis of sharp medical instrument injuries]. PMID- 19309600 TI - [A successful case of treatment of high-dosage mercuric sulfide poisoning]. PMID- 19309601 TI - [Current state of digestive system robotic surgery in the light of evidence based medicine]. AB - The incorporation of robotics in minimally invasive surgery has had mixed reception in the different fields of digestive surgery. Nowadays we are exposed to a continuous stream of publications on robotic approach techniques and outcomes, which do not always provide objective criteria and whose value, through scientific evidence analysis, is sometimes arguable. With the aim of shedding light on current knowledge on digestive robotic surgery and giving an update of its possibilities, the authors analyse the abundant literature available on the different digestive robotic surgery procedures, and sum up their own experience. PMID- 19309602 TI - [Leiomyoma of the oesophagus]. AB - INTRODUCTION: Oesophageal leiomyoma is a rare tumour. The purpose of this paper is to describe the form of presentation, the approaches and the surgical technique, in five consecutive cases. MATERIAL AND METHOD: A retrospective analysis was performed on the data recorded prospectively on five consecutive patients operated on between 2003 and 2007. RESULTS: All the patients were seen due to dysphagia to solids. All were examined with contrast radiology, video endoscopy and computed tomography, although 4 of them also had ultrasound endoscopy performed. The surgical technique was extra-mucosal enucleation, with 3 cases by thoracoscopy and 2 by thoracotomy. There were no complications. The immunohistochemistry study was negative for CD117 and CD34 in all cases. CONCLUSIONS: Enucleation of oesophageal leiomyomas by video-thoracoscopy is safe, well tolerated, and there is a rapid recovery, and is the procedure of choice for this benign tumour. Thoracotomy may be necessary in some presentations and locations. PMID- 19309603 TI - [Results of a study on populational colorectal cancer screening]. AB - INTRODUCTION: As colorectal cancer (CRC) screening based on occult blood detection has been shown to be effective in reducing mortality due to this disease, it is now important to decide on the best methods to obtain the maximum numbers of participants. The aim of the study was to analyse the results from a pilot CRC screening programme in a general population sample in Barcelona. A follow-up of false positive cases was made after five years. PATIENTS AND METHOD: A cross section of the population aged 50-74 years in one primary health care centre was studied. The screening test consisted of an immunological method for the detection of faecal occult blood which was sent to the homes of the target population. RESULTS: Participation was 46.6%, 11.7% of the tests were positive, and 79.3% agreed to have a colonoscopy. Eight adenocarcinomas and 32 patients with adenomas >0.4 cm were diagnosed. CONCLUSIONS: The results obtained on the initial participation and the follow-up at five years suggest the viability of a CCR screening programme in our country. PMID- 19309604 TI - [Prosthetic infection after hernioplasty. Five years experience]. AB - INTRODUCTION: Prosthesis infection is an infrequent but important complication in abdominal wall surgery. The aim of this study is to evaluate the incidence and risk factors for the infection of the prosthesis after hernia repair, as well as the treatment to apply. MATERIAL AND METHOD: Between January 2002 and December 2006, we performed 1055 prosthetic hernia repairs: 761 inguinal hernias (72.1%), 74 umbilical hernias (7%) and 220 ventral hernias (20.9%). We prospectively analysed preoperative, intraoperative and postoperative variables, as well as the incidence of infection of surgical wound and of prosthesis. We used ASA classification for preoperative anaesthetic evaluation. RESULTS: The overall percentage of infection of the prosthesis was 1.3%. Infection was observed in 11 repairs with polypropylene mesh (PPL), in 4 with PTFE mesh, and one case in combined mesh. Risk factors of mesh infection were: obesity (p=0.002), diabetes (p=0.020), the type of repair (p=0.047), emergency surgery (p=0.001), the type and size of mesh (p=0.003; p=0.007) and time of surgery >180 min (p<0.001). Seven of the 11 patients with infection of PPL prosthesis were resolved with conservative treatment, whereas all the cases with PTFE infection or mixed mesh needed removal to solve the problem. CONCLUSIONS: Several factors are involved in producing a prosthesis infection. Whereas antibiotic treatment and surgical drainage of the infection can be sufficient in most PPL mesh infection, PTFE prostheses need to be removed prematurely in order to halt the infection process. PMID- 19309605 TI - [Mucinous cystadenoma of the pancreas]. PMID- 19309606 TI - A double-blind, prospective, randomised, controlled clinical trial of minimally invasive dynamic hip screw fixation of intertrochanteric fractures. AB - OBJECTIVE: To compare minimally invasive dynamic hip screw (MIDHS) fixation with conventional dynamic hip screw (CDHS) fixation for treatment of intertrochanteric femoral fracture. METHODS: Of the 66 participants in this double-blind study, 35 were randomised to MIDHS and 31 to CDHS fixation. Main outcome measurements were wound size, haemoglobin decrease, blood transfusion rate, pain score, analgesic consumption, Elderly Mobility Scale score, hip screw position, tip-apex distance, union rate, time to healing and complication rate. RESULTS: The groups had similar preoperative clinical data. Postoperatively the MIDHS group had significantly smaller wound size, less blood loss, lower blood transfusion rates, pain scores and rates of analgesic consumption, and higher early Elderly Mobility Scale scores. There were no significant differences in fracture alignment, hip screw position, tip-apex distance, union rate, time to healing or complication rate. CONCLUSION: MIDHS fixation of intertrochanteric fractures is effective and safe and significantly reduces blood loss, pain and rehabilitation period, without sacrificing reduction alignment, screw position, fixation stability or bone healing. PMID- 19309637 TI - When is being in a hurry going too fast? Lessons learned from clinical trials in cervical cancer. PMID- 19309638 TI - Hereditary breast and ovarian cancer syndrome. PMID- 19309639 TI - Emerging techniques in aesthetic plastic surgery. Preface. PMID- 19309640 TI - Emerging techniques in aesthetic plastic surgery. AB - This article outlines the experience of authorities on emerging techniques in plastic surgery that are discussed within this issue, such as mesotherapy and lipodissolve, Russian threads, radiofrequency, and "laser lipo." Readers are invited to analyze the findings and determine whether there is something that could apply to their practices. PMID- 19309641 TI - Mesotherapy and injection lipolysis. AB - The first part of this article familiarizes the reader with the evolution of mesotherapy, injection lipolysis, and the use of phosphatidylcholine and deoxycholate for subcutaneous fat reduction. There is an emphasis on the underlying basic science of fat metabolism and the biochemistry of phosphatidylcholine, so that practitioners will be able to understand future published research on these topics. The second half details some personal experience with injection lipolysis. PMID- 19309643 TI - Refinement of technique in injection lipolysis based on scientific studies and clinical evaluation. AB - The unusual evolution of the practice of injection lipolysis has generated doubt regarding its safety and efficacy among many physicians. During the early years of this decade, mesotherapy was practiced by a few physicians, but the practice was not widespread. Paramedical practitioners and business developers saw the market potential for nonsurgical fat reduction, and the practice of injection lipolysis was packaged and sold before the mechanism of action was understood. Because of the early lack of scientific research and understanding of the limitations of injection lipolysis, many unsuitable patients were treated with this modality. To better understand the way injection lipolysis works, the inclusion and exclusion criteria for patients desiring treatment, and an accurate clinical evaluation format for potential treatment regions, a series of scientific studies was performed in 2007 and early 2008. These studies included a serial histopathology evaluation of treated patients over time, a stem cell study performed with the McGowan Research Institute in Pittsburgh, an animal study performed in conjunction with the Colorado State University veterinary school, and a prospective multicenter clinical trial using injection lipolysis in the back roll region. The purpose of these studies was to determine the way injection lipolysis works, how modifications of the formula and technique change the outcome, the role of each constituent component of various formulas, and the degree of fat reduction and skin retraction that is attainable with these treatments. The influence of depth of injection, distance between injection points, volume of injection, and ratios of constituent components was studied. The degree of topographic contour correction and the amount of volume reduction were evaluated. Following a review of these recent studies, an updated recommendation for the clinical practice of injection lipolysis was formulated. PMID- 19309645 TI - The lipodissolve technique: clinical experience. AB - This article describes the author's experience with lipodissolve, a nonsurgical treatment to reduce fat deposits via the injection of a single component, phosphatidylcholine, a natural product found in the body. Lipodissolve is a relatively new aesthetic procedure used to dissolve smallish, localized, and defined zones of fat in the face and body. PMID- 19309648 TI - Surgisis acellular collagen matrix in aesthetic and reconstructive plastic surgery soft tissue applications. AB - Tissue engineering in aesthetic and reconstructive plastic surgery remains an elusive goal. The advent of Surgisis extracellular collagen matrix and its performance characteristics suggest that the use of a bioengineered tissue substitute can meet some of our reconstructive requirements. Incorporation and replacement by host tissue with minimal allergic or immune response seems to be achievable today. The ability to engineer the device, the ready availability of substrate, and its cost effectiveness support the use of Surgisis in aesthetic and reconstructive plastic surgery applications. Future product innovations and engineering seem promising. The permanent role of Surgisis in aesthetic and reconstructive plastic surgery will be determined by its documented long-term performance. PMID- 19309649 TI - Laser-assisted liposuction. AB - In the United States, as in many other countries, liposuction is the most commonly performed cosmetic surgical procedure. Advances in technology have enabled surgeons to improve the safety and efficacy of the procedure. One such technological advance is laser-assisted liposuction. This minimally invasive technique employs laser energy in direct contact with adipose tissue to induce lipolysis and, at the same time, coagulate tiny blood vessels and stimulate dermal and subdermal neocollagenesis. These features of laser lipolysis permit a fast, comfortable postoperative recovery, a rapid return to activities of daily living, and excellent skin redraping as a result of laser-induced skin tightening. PMID- 19309652 TI - Clinical applications of radiofrequency: nonsurgical skin tightening (thermage). AB - Thermage is a nonsurgical treatment for sagging skin in the jowl, neck, and eyelids. Abdominal striae, loose upper arm skin and buttock sagging, and improvement of the appearance of cellulite have become amenable to the radiofrequency treatment. Radiofrequency passed through cooled epidermis allows for radiofrequency-induced thermal damage to the dermis and deep dermal collagen (fibrous septae). The remodeling of the collagen leads to clinically discernible improvement in the sagging skin and skin quality; the remodeling of deep dermal collagen (fibrous septae) allows modeling of contours and improvement of the cellulite appearance. PMID- 19309653 TI - Breast reconstruction and augmentation using pre-expansion and autologous fat transplantation. AB - The concept of fat grafting for volume enhancement is not a new one. There is much to study to maximize both graft volume and, more importantly, patient safety. This article outlines the authors' approach to breast deformities using fat grafting, with emphasis on current technique. PMID- 19309654 TI - APTOS suture lifting methods: 10 years of experience. AB - Over the past decade we have developed and successfully used methods of minimally invasive aesthetic surgery, based on special threads provided with microscopic angled barbs (Aptos Thread), double-pointed needles (Aptos Needle), and elastic needles (Aptos Spring). This article describes our original techniques of subcutaneous suturing through and lifting of soft tissue, which make it possible to obtain qualitative, sufficiently long-standing lifting of the face and neck with no cuts or with tiny cutaneous incisions. Our 10-year experience in using the Aptos methods has demonstrated that these are simple, safe, and efficient techniques for rejuvenation of the face and neck. PMID- 19309680 TI - Predictive values of the anal sphincter electromyography. PMID- 19309655 TI - A new approach for the prophylactic improvement of surgical scarring: avotermin (TGF beta 3). AB - Patients and physicians are concerned about scarring resulting from surgery, and patients, in particular, value even small improvements in scarring. Translational research into the processes involved in scarring at the molecular, cellular, and tissue levels has facilitated the discovery and development of new biological approaches for improving scarring. This article highlights research concerning avotermin (human recombinant TGF beta 3), the first in a new class of prophylactic medicines that may promote the regeneration of normal skin and improve scar appearance. PMID- 19309681 TI - Does being an orphan decrease the nutritional status of Luo children? AB - The HIV/AIDS pandemic is creating a generation of orphaned children in Africa. The number of orphans will continue to increase long after the HIV/AIDS crisis has peaked; therefore, it is important to determine how best to assist these children. Current studies investigating the impact of orphanhood have conflicting results and conclusions. Several studies report that orphans are at a disadvantage and are more likely to suffer from malnutrition, whereas other studies report no difference between the nutritional status of orphans and nonorphans. Four hundred eleven Luo children (mean age 9 +/- 1 years) were recruited to participate in a study investigating the impact of orphanhood on nutritional status. Participants underwent an interview, anthropometric measurements, testing for anemia, a clinical history, and a physical exam. Anthropometric variables and hemoglobin level were compared across groups using a t-test. The reference population used for comparison of anthropometric variables is the 2000 CDC growth reference data. All analyses were gender specific, and the effect of length of orphanhood was also investigated. The data presented here suggest that there is no significant difference between the nutritional status of orphaned and nonorphaned Luo children. This study supports research indicating there is little, if any, difference in nutritional indicators between orphans and nonorphans. Orphans may live in households with higher socioeconomic statuses than nonorphans. Luo orphans may not be at higher risk for poor nutritional status than nonorphans; therefore, interventions targeted at this age group should include both orphaned and nonorphaned children. PMID- 19309682 TI - Maturity-related differences in physical activity among 10- to 12-year-old girls. AB - Besides environmental and psycho-social factors explaining the variation in physical activity levels during adolescence, some evidence suggests that biological processes are involved in regulating habitual daily physical activity and energy expenditure. The purpose of this study was to examine the influence of biological maturity status on physical activity. Chronological age, standing height, sitting height, and body mass were measured cross-sectionally in 268 girls, aged 9.5 to 11.5 years. Biological maturity groups (Early, Average, Late) were created according to estimated age at peak-height-velocity (estAPHV). Habitual physical activity was determined with a pedometer (Yamax Digiwalker SW 200) over a 7-day period. Differences in steps/day across maturity groups were examined by ANCOVA, controlling separately for time the pedometer was worn, leg length, and body mass. Mean pedometer steps/day was 10,822 +/- 2,639. As expected, body size varied by maturity status (e.g., early > average > late). Significant maturity group differences were found with early maturing girls showing lower activity levels compared to average or late maturers. These differences remained after controlling for time the pedometer was worn and leg length; however, the differences were no longer significant when controlling for body mass. The results suggest that biological maturity status influences physical activity levels in girls between 10 and 12 years of age but the relationship is not independent of body mass. Further research is needed to establish the complex inter-relationships among adiposity, biological maturation, and energy expenditure during puberty. PMID- 19309683 TI - A geometric morphometric approach to the quantification of population variation in sub-Saharan African crania. AB - We report here on new data examining cranial variation in 18 modern human sub Saharan African populations. Previously, we investigated variation within southern Africa; we now extend our analyses to include a series of Central, East, and West African crania, to further knowledge of the relationships between, and variation and regional morphological patterning in, those populations. The sample comprises 377 male individuals; the three-dimensional coordinates of 96 landmarks are analyzed using Procrustes-based methods. Interpopulation variation is examined by calculating shape distances between groups, which are compared using resampling statistics and parametric tests. Phenotypic variance, as a proxy for genetic variance, is measured and compared across populations. Principal components and cluster analyses are employed to explore relationships between the populations. Shape differences are visualized using three-dimensional rendered models. Observed disparity patterns imply a mix of differences and similarities across populations, with no apparent support for genetic bottlenecks, which is likely a consequence of migrations that may have influenced differences in cranial form; supporting data are found in recent molecular studies. The Pygmy sample had the most distinctive cranial morphology; characteristically small in size with marked prognathism. These features characterized, although less strongly, the neighboring Bateke, and are possibly related to similar selective pressures in conjunction with interbreeding. Small cranial size is also involved in the considerable distinctiveness of the San and Khoikhoi. The statistical procedures applied in this study afford a powerful and robust means of quantifying and visualizing the magnitude and pattern of cranial variation between sub-Saharan African populations. PMID- 19309684 TI - Short- and long-term consequences of early parental loss in the historical population of the Krummhorn (18th and 19th century). AB - The impact of the early loss of one's father or one's mother on the survival and age at death of children was investigated on the basis of a historical reconstitution of families from the Krummhorn (East Frisia/Ostfriesland; Germany) with the aid of Kaplan-Meier plots and the Cox regression. In our analyses, we took into account the changed situation of the family after the death of a parent by incorporating the surviving spouse's remarriage or relationships with stepparents. We find that the impact on survival of the children was sex-specific and also depended on whether and at what point in time during childhood their father or mother had died. As expected, children's immediate survival was strongly affected by maternal loss. A few results can be construed as survival diminishing long-term consequences of the early loss of a parent. Daughters who lost their fathers before their first birthday proved to have increased mortality over a longer period of their youth. The age at death of daughters was also lowered if they had to live with a step-mother during early childhood. To interpret these results, three hypotheses, including an (intrinsic) trade-off, compensation and a selection scenario, were tested. Other approaches, which are based, for example, on the extrinsic trade-off between mating effort and parental investment of the surviving parent, also appear to be suitable as an explanation for the long-term consequences, which eventually draws the conclusion that the compensation scenario is the most likely explanation for the consequences of early parental loss. PMID- 19309686 TI - A novel topology for representing protein folds. AB - Various topologies for representing 3D protein structures have been advanced for purposes ranging from prediction of folding rates to ab initio structure prediction. Examples include relative contact order, Delaunay tessellations, and backbone torsion angle distributions. Here, we introduce a new topology based on a novel means for operationalizing 3D proximities with respect to the underlying chain. The measure involves first interpreting a rank-based representation of the nearest neighbors of each residue as a permutation, then determining how perturbed this permutation is relative to an unfolded chain. We show that the resultant topology provides improved association with folding and unfolding rates determined for a set of two-state proteins under standardized conditions. Furthermore, unlike existing topologies, the proposed geometry exhibits fine scale structure with respect to sequence position along the chain, potentially providing insights into folding initiation and/or nucleation sites. PMID- 19309687 TI - Aegerolysins: structure, function, and putative biological role. AB - Aegerolysins, discovered in fungi, bacteria and plants, are highly similar proteins with interesting biological properties. Certain aegerolysins possess antitumoral, antiproliferative, and antibacterial activities. Further possible medicinal applications include their use in the prevention of atherosclerosis, or as vaccines. Additional biotechnological value of fungal aegerolysins lies in their involvement in development, which could improve cultivation of commercially important edible mushrooms. Besides, new insights on microheterogeneity of raft like membrane domains could be gained by using aegerolysins as specific markers in cell and molecular biology. Although the exact function of aegerolysins in their producing organisms remains to be explained, they are biochemically well characterized all-beta structured proteins sharing the following common features: low isoelectric points, similar molecular weights (15-17 kDa), and stability in a wide pH range. PMID- 19309688 TI - PORCN mutations in focal dermal hypoplasia: coping with lethality. AB - The X-linked dominant trait focal dermal hypoplasia (FDH, Goltz syndrome) is a developmental defect with focal distribution of affected tissues due to a block of Wnt signal transmission from cells carrying a detrimental PORCN mutation on an active X-chromosome. Molecular characterization of 24 unrelated patients from different ethnic backgrounds revealed 23 different mutations of the PORCN gene in Xp11.23. Three were microdeletions eliminating PORCN and encompassing neighboring genes such as EBP, the gene associated with Conradi-Hunermann-Happle syndrome (CDPX2). 12/24 patients carried nonsense mutations resulting in loss of function. In one case a canonical splice acceptor site was mutated, and 8 missense mutations exchanged highly conserved amino acids. FDH patients overcome the consequences of potentially lethal X-chromosomal mutations by extreme skewing of X-chromosome inactivation in females, enabling transmission of the trait in families, or by postzygotic mosaicism both in male and female individuals. Molecular characterization of the PORCN mutations in cases diagnosed as Goltz syndrome is particularly relevant for genetic counseling of patients and their families since no functional diagnostic test is available and carriers of the mutation might otherwise be overlooked due to considerable phenotypic variability associated with the mosaic status. PMID- 19309689 TI - Genetic and epigenetic analysis of recurrent hydatidiform mole. AB - Familial biparental hydatidiform mole (FBHM) is a maternal-effect autosomal recessive disorder in which recurrent pregnancy failure with molar degeneration occurs. The phenotype mimics molar pregnancy due to androgenesis, despite the normal genetic makeup of the conceptus. FBHM appears to result from a failure to establish correct maternal epigenetic identity at imprinted loci during oogenesis. Several women affected with FBHM have previously been shown to have biallelic mutations in the NLRP7 gene (NALP7). Here, we present the results of epigenetic and mutational analysis on FBHM patients from 11 families, 10 of them novel. We demonstrate a methylation defect at imprinted loci in tissue from four new FBHM cases. Biallelic NLRP7 mutations, including eight previously undescribed mutations, were found in all but one family. These results indicate for the first time that maternal imprints at some loci may be correctly specified in FBHM conceptions, since differential methylation of SGCE/PEG10 was preserved in all four cases. PMID- 19309690 TI - Copy number variation at the FCGR locus includes FCGR3A, FCGR2C and FCGR3B but not FCGR2A and FCGR2B. AB - Human Fcgamma receptors (FcgammaRs) are glycoproteins that bind the Fc region of IgG. The genes encoding the low-affinity FcgammaRs are located on chromosome 1q23 24. Beside single nucleotide polymorphisms (SNPs), gene copy number variation (CNV) is now being recognized as an important indicator for inter-individual differences. Recent studies on identifying CNV in the human genome suggest large areas at chromosome 1q23-24 to be involved, and CNV in this region has been associated with manifestations of systemic autoimmune disease. To study both SNPs and CNV of the low-affinity FcgammaRs in one assay, we have developed a Multiplex Ligation-dependent Probe Amplification (MLPA) assay. A novel CNV for FCGR3A was observed. Similar to FCGR3B and FCGR2C, a gene-dosage effect of FCGR3A was found, that seemed to correlate nicely with the FcgammaRIIIa expression on NK cells. Next, we delineated the approximate boundaries of CNV at the FCGR locus. Variation in co-segregation of neighboring FCGR genes was limited to four variants, with patterns of Mendelian inheritance. No CNV of the FCGR2A and FCGR2B genes was observed in over 600 individuals. In conclusion, we report a novel CNV of the FCGR3A gene that correlates with FcgammaRIIIa expression and function on NK cells. Only FCGR3A, FCGR2C and FCGR3B show CNV, in contrast to FCGR2A and FCGR2B. PMID- 19309691 TI - Retention of lysosomal protein CLN5 in the endoplasmic reticulum causes neuronal ceroid lipofuscinosis in Asian sibship. AB - The neuronal ceroid lipofuscinoses (NCLs) form a group of autosomal recessively inherited neurodegenerative disorders that mainly affect children. Ten NCL forms can be distinguished by age at onset, clinicopathologic features, and genetics. In eight of these forms, the underlying genes have been identified. At present, approximately 10% of all patients do not fall into one of the eight known genetic forms of NCL. We have identified two Asian families with two novel homozygous mutations in the CLN5 gene. In the first Pakistani family, two children developed symptoms of an early juvenile NCL. After exclusion of mutations in genes known to be associated with this age of onset in families from many different countries (CLN1, CLN2, CLN3, CLN6, CLN8 and CLN10) SNP array-based homozygosity mapping led to the identification of a novel homozygous mutation c.1072_1073delTT (p.Leu358AlafsX4) in CLN5. In the second Afghan family, two children developed symptoms of a late infantile NCL. The mutation c.1137G>T (p.Trp379Cys) in CLN5 was identified. The affected children in these families represent the first reported CLN5 patients originating in Asian sibships. Expression analysis showed that mutant p.Leu358AlafsX4 CLN5 is truncated and lacks a used N-glycosylation site at Asn401. The missense mutation p.Trp379Cys affected neither the size nor glycosylation of the CLN5 protein. Double immunofluorescence microscopy showed that while the wild-type CLN5 protein is localized in lysosomes, both mutant CLN5 proteins are retained in the endoplasmic reticulum rather than reaching the lysosome. PMID- 19309692 TI - Identification and characterization of novel collagen VI non-canonical splicing mutations causing Ullrich congenital muscular dystrophy. AB - Splicing mutations occurring outside the invariant GT and AG dinucleotides are frequent in disease genes and the definition of their pathogenic potential is often challenging. We have identified four patients affected by Ullrich congenital muscular dystrophy and carrying unusual mutations of COL6 genes affecting RNA splicing. In three cases the mutations occurred in the COL6A2 gene and consisted of nucleotide substitutions within the degenerated sequences flanking the canonical dinucleotides. In the fourth case, a genomic deletion occurred which removed the exon8-intron8 junction of the COL6A1 gene. These mutations induced variable splicing phenotypes, consisting of exon skipping, intron retention and cryptic splice site activation/usage. A quantitative RNA assay revealed a reduced level of transcription of the mutated in-frame mRNA originating from a COL6A2 point mutation at intronic position +3. At variance, the transcription level of the mutated in-frame mRNA originating from a genomic deletion which removed the splicing sequences of COL6A1 exon 8 was normal. These findings suggest a different transcriptional efficiency of a regulatory splicing mutation compared to a genomic deletion causing a splicing defect. PMID- 19309693 TI - Phenotypic spectrum of STRA6 mutations: from Matthew-Wood syndrome to non-lethal anophthalmia. AB - Matthew-Wood, Spear, PDAC or MCOPS9 syndrome are alternative names used to refer to combinations of microphthalmia/anophthalmia, malformative cardiac defects, pulmonary dysgenesis, and diaphragmatic hernia. Recently, mutations in STRA6, encoding a membrane receptor for vitamin A-bearing plasma retinol binding protein, have been identified in such patients. We performed STRA6 molecular analysis in three fetuses and one child diagnosed with Matthew-Wood syndrome and in three siblings where two adult living brothers are affected with combinations of clinical anophthalmia, tetralogy of Fallot, and mental retardation. Among these patients, six novel mutations were identified, bringing the current total of known STRA6 mutations to seventeen. We extensively reviewed clinical data pertaining to all twenty-one reported patients with STRA6 mutations (the seven of this report and fourteen described elsewhere) and discuss additional features that may be part of the syndrome. The clinical spectrum associated with STRA6 deficiency is even more variable than initially described. PMID- 19309694 TI - How long does a mindfulness-based stress reduction program need to be? A review of class contact hours and effect sizes for psychological distress. AB - The mindfulness-based stress reduction (MBSR) program was designed to be long enough for participants to grasp the principles of self-regulation through mindfulness and develop skill and autonomy in mindfulness practice. It traditionally consists of 26 hours of session time including eight classes of 2 1/2 hours and an all-day class. The circumstances of some groups exclude them from participating in this standard form and a number of trials have evaluated programs with abbreviated class time. If lower program time demands can lead to similar outcomes in psychological functioning, it would support their utility in these settings and might lead to greater participation. However, the effect of variation in class hours on outcomes has not been systematically studied. To obtain preliminary information related to this question we examined effect sizes for psychological outcome variables in published studies of MBSR, some of which had adapted the standard number of class hours. The correlation between mean effect size and number of in-class hours was nonsignificant for both clinical and nonclinical samples and suggests that adaptations that include less class time may be worthwhile for populations for whom reduction of psychological distress is an important goal and for whom longer time commitment may be a barrier to their ability or willingness to participate. However, the standard MBSR format has accrued the most empirical support for its efficacy and session time may be important to the development of other kinds of program outcomes. The result points to the importance of empirical studies systematically examining this question. PMID- 19309695 TI - Computational studies of H5N1 influenza virus resistance to oseltamivir. AB - Influenza A (H5N1) virus is one of the world's greatest pandemic threats. Neuraminidase (NA) inhibitors, oseltamivir and zanamivir, prevent the spread of influenza, but drug-resistant viruses have reduced their effectiveness. Resistance depends on the binding properties of NA-drug complexes. Key residue mutations within the active site of NA glycoproteins diminish binding, thereby resulting in drug resistance. We performed molecular simulations and calculations to characterize the mechanisms of H5N1 influenza virus resistance to oseltamivir and predict potential drug-resistant mutations. We examined two resistant NA mutations, H274Y and N294S, and one non-drug-resistant mutation, E119G. Six nanosecond unrestrained molecular dynamic simulations with explicit solvent were performed using NA-oseltamivir complexes containing either NA wild-type H5N1 virus or a variant. MM_PBSA techniques were then used to rank the binding free energies of these complexes. Detailed analyses indicated that conformational change of E276 in the Pocket 1 region of NA is a key source of drug resistance in the H274Y mutant but not in the N294S mutant. PMID- 19309696 TI - In vivo validation of CAAS QCA-3D coronary reconstruction using fusion of angiography and intravascular ultrasound (ANGUS). AB - OBJECTIVES: The CAAS QCA-3D system (Pie Medical Imaging BV, the Netherlands) was validated against 3D reconstructions based on fusion of angiography and intravascular ultrasound (ANGUS), allowing slice by slice validation of the lumen areas and 3D geometric values. BACKGROUND: Accurate online 3D reconstruction of human coronary arteries is of outmost importance during clinical practice in the catheterization laboratory. The CAAS QCA-3D system provides technology to 3D reconstruct human coronary arteries based on two or more angiographic images, but was not validated in realistic arteries before. METHODS: Ten patients were imaged using biplane angiography and an ECG gated (TomTec) intravascular ultrasound (IVUS) pullback (stepsize 0.5 mm, Boston Scientific). The coronary arteries were 3D reconstructed based on (a) fusion of biplane angiography and IVUS (ANGUS) and (b) CAAS QCA-3D using the biplane angiography images. For both systems the length, the curvature and the lumen areas at 0.5 mm spacing were calculated and compared. RESULTS: Bland-Altman analysis indicated that the CAAS QCA-3D system underestimated the lumen areas systematically by 0.45 +/- 1.49 mm2. The segment length was slightly underestimated by the CAAS QCA-3D system (62.1 +/- 11.3 vs. 63.2 +/- 11.4 mm; P < 0.05), while the curvature of the analyzed segments were not statistically different. CONCLUSIONS: The CAAS QCA-3D system allows 3D reconstruction of human coronary arteries based on biplane angiography. Validation against the ANGUS system showed that both the 3D geometry and lumen areas are highly correlated which makes the CAAS QCA-3D system a promising tool for applications in the catheterization laboratory and opens possibilities for computational fluid dynamics. PMID- 19309697 TI - Structural characterization of unphosphorylated STAT5a oligomerization equilibrium in solution by small-angle X-ray scattering. AB - Signal transducer and activator of transcription (STAT) proteins play a crucial role in the activation of gene transcription in response to extracellular stimuli. The regulation and activity of these proteins require a complex rearrangement of the domains. According to the established models, based on crystallographic data, STATs convert from a basal antiparallel inactive dimer into a parallel active one following phosphorylation. The simultaneous analysis of small-angle X-ray scattering data measured at different concentrations of unphosphorylated human STAT5a core domain unambiguously identifies the simultaneous presence of a monomer and a dimer. The dimer is the minor species but could be structurally characterized by SAXS in the presence of the monomer using appropriate computational tools and shown to correspond to the antiparallel assembly. The equilibrium is governed by a moderate dissociation constant of K(d) approximately 90 microM. Integration of these results with previous knowledge of the N-terminal domain structure and dissociation constants allows the modeling of the full-length protein. A complex network of intermolecular interactions of low or medium affinity is suggested. These contacts can be eventually formed or broken to trigger the dramatic modifications in the dimeric arrangement needed for STAT regulation and activity. PMID- 19309698 TI - Engineering a G protein-coupled receptor for structural studies: stabilization of the BLT1 receptor ground state. AB - Structural characterization of membrane proteins is hampered by their instability in detergent solutions. We modified here a G protein-coupled receptor, the BLT1 receptor of leukotriene B(4), to stabilize it in vitro. For this, we introduced a metal-binding site connecting the third and sixth transmembrane domains of the receptor. This modification was intended to restrain the activation-associated relative movement of these helices that results in a less stable packing in the isolated receptor. The modified receptor binds its agonist with low-affinity and can no longer trigger G protein activation, indicating that it is stabilized in its ground state conformation. Of importance, the modified BLT1 receptor displays an increased temperature-, detergent-, and time-dependent stability compared with the wild-type receptor. These data indicate that stabilizing the ground state of this GPCR by limiting the activation-associated movements of the transmembrane helices is a way to increase its stability in detergent solutions; this could represent a forward step on the way of its crystallization. PMID- 19309699 TI - A new 0.010-inch guidewire and compatible balloon catheter system: the IKATEN registry. AB - OBJECTIVE: To evaluate the safety and feasibility of a new 0.010-inch guidewire and a specialized balloon catheter for the 0.010-inch guidewire in routine percutaneous coronary intervention (PCI). BACKGROUND: Several reports have shown that a new 0.010-inch system is effective for specific situations where reduction of catheter size may be necessary. However, the safety of this system in routine PCI is unknown. METHODS: The IKATEN registry is a prospective, multicenter, nonrandomized registry study. Patients who underwent elective PCI with a 0.010 inch guidewire and its associated balloon catheter as primary devices were enrolled. The coprimary endpoints were clinical success and device success rates. The secondary endpoints were major adverse cardiac events (MACE) and bleeding complications. RESULTS: A total of 133 patients with 148 lesions were enrolled. The majority were male (75.3%), and mean age was 68 +/- 10 years. Type B2/C lesions comprised 60% of the lesions, prevalence of chronic total occlusion (CTO) was 16.9%, and bifurcation lesions were found in 22.3% of patients. A transradial approach was used in 79.7% of patients, and the average guiding catheter size was 5.1 +/- 0.4 Fr. Clinical success rate was 99.2%, and device success rate was 99.3%. Device failure occurred only in one case of chronic total occlusion because of unsuccessful guidewire passage. No MACE or bleeding complications were reported except for a small hematoma at the puncture site in one patient. Stent delivery success rate on 0.010-inch guidewire was 93.9% because of failure of stent balloon to pass eight lesions. CONCLUSION: The IKATEN registry data suggest that the 0.010-inch system is safe and its use is feasible in routine PCI including bifurcation and CTO lesions. PMID- 19309700 TI - Prognostic value of the Syntax score in patients undergoing coronary artery bypass grafting for three-vessel coronary artery disease. AB - BACKGROUND: The Syntax score prognostic value has recently been highlighted in patients undergoing percutaneous coronary intervention (PCI) for multivessel coronary artery disease (CAD), however its prognostic value in patients undergoing coronary artery bypass grafting (CABG) for multivessel CAD is still unknown. The aim of this study was to evaluate the prognostic value of the Syntax score in patients undergoing CABG for 3-vessel CAD. METHODS: A cohort of 320 consecutive patients with multivessel (3-vessel) CAD who were subjected for CABG were included in this study and divided into tertiles according to the Syntax score (<24.5, 24.5-34, and >34). During the 1-year follow-up, cardiovascular events including death, myocardial infarction (MI), and stroke were systematically indexed. The primary end point was the composite criteria death/MI/stroke. RESULTS: The Syntax score ranged from 11-74 with a mean of 31.2 +/- 12.6 and a median of 28.5 [22-38]. Baseline clinical characteristics were similar among the tertiles. No statistical difference was found for the composite criteria death/MI/stroke: 9.4% versus 7.5% versus 10.4% in the groups with a Syntax score <24.5, 24.5-34, and >34, respectively (P = 0.754). CONCLUSION: Unlike for PCI, the Syntax score has a poor prognostic value for severe cardiovascular events in patients undergoing CABG for 3-vessel CAD. Other risk scores should be used to predict the outcome of this population. PMID- 19309701 TI - Inadvertent subclavian artery catheter placement complicated by stroke: endovascular management and review. AB - BACKGROUND: Removal of large-bore venous catheters misplaced within the subclavian artery risks significant hemorrhage due to the noncompressible entry site. Comorbidities in these patients often make traditional surgical approaches prohibitive. CASE DESCRIPTION: We present a case of inadvertent subclavian artery placement of a central venous catheter complicated by thromboembolic stroke, illustrate treatment with a combination of percutaneous closure and temporary balloon tamponade, and review the endovascular management of this unusual problem. CONCLUSION: Endovascular treatment of inadvertent subclavian artery catheterization appears to be a reasonable option, and its less-invasive nature may be beneficial for patients in whom the underlying disease initially required central venous access. PMID- 19309702 TI - The need for a dedicated bifurcation stenting system. PMID- 19309703 TI - Is the glass 97% full, or 3% empty? Reinfarction and stent thrombosis after STEMI. PMID- 19309704 TI - A third dimension to uncloud the funnel plot of operator quality. PMID- 19309705 TI - Transcatheter closure of adults with atrial septal defect and atrial fibrillation -the right thing to do. PMID- 19309706 TI - Alternatives to the pull and hope technique to inadvertent subclavian artery puncture. PMID- 19309707 TI - Which came first? The wire or the balloon? Looking ahead. PMID- 19309708 TI - The SYNTAX score in practice: an aid for patient selection for complex PCI. PMID- 19309709 TI - Clinical follow-up in endovascular treatment for TASC C-D lesions in femoro popliteal segment. AB - OBJECTIVE: To demonstrate the technical success and clinical follow-up after endovascular treatment of femoropopliteal segment TASC II C and D lesions. METHODS: From July 2002 to February 2007, 234 limbs in 190 patients with femoropopliteal segment TASC II C (n = 112) and D (n = 122) lesions were treated. Endovascular treatment consisted of PTA, fibrinolysis and PTA, subintimal recanalization and PTA, and finally stent graft. Patients were clinically evaluated at 30 days, 3, 6 month, and at 1 year in the outpatient setting with clinical examination and ankle-brachial indices (ABI). In the case of stent placement, additional ultrasound evaluation was performed at 12, 24, and 48 month. RESULTS: 49.5% of procedures were performed on patients with lifestyle limiting claudication (IC) and 50.5% were performed for critical limb ischemia (CLI). Technical success, defined as successful recanalization and treatment of the occluded vessel, was achieved in 97% of cases. Periprocedural mortality was 3.15% and all deaths occurred in the CLI group. A follow-up 13 +/- 6 months and was achieved in 76%. During the follow-up, clinical outcome for IC group and clinical CLI group was asymptomatic 72% vs. 29.8%, symptomatic with clinical improvement 22% vs. 33.7%, and major amputation 3% vs. 23.3%. CONCLUSION: The majority of claudicating patients with femoropopliteal TASC II C and D lesions will benefit from the endovascular treatment. Patient presenting CLI have a worse outcome, nevertheless the endovascular treatment can delay amputation, preserving the native vessel and does not impede surgical bypass if needed. For this reason, we consider that endovascular treatment may be the first choice treatment even in femoropopliteal TASC II C and D lesions. PMID- 19309710 TI - Oh say can you C(T)? PMID- 19309711 TI - We are ready, willing, and able: the interventional cardiology community: but we have two hands tied behind our back. PMID- 19309712 TI - Incidence, predictors, and outcome of reinfarction and stent thrombosis within one year after primary percutaneous coronary intervention for ST-elevation myocardial infarction. AB - BACKGROUND: Reinfarction and stent thrombosis are major complications after primary percutaneous coronary intervention (PCI) for ST-elevation myocardial infarction (STEMI). OBJECTIVE: We sought to investigate the incidence, predictors, and outcome of reinfarction and stent thrombosis in a contemporary cohort of STEMI patients. METHODS: Reinfarction and stent thrombosis within 1 year after primary PCI for STEMI were analyzed in the Thrombus Aspiration during Percutaneous coronary intervention in Acute myocardial infarction Study (TAPAS). RESULTS: Reinfarction was observed in 3.4% (34/995) of patients within 1 year after primary PCI. Angiographic evidence of stent thrombosis was observed in 15/34 (44.1%). During the index primary PCI, intra-aortic balloon counterpulsation was used in 10/32 (31.3%) patients with and 47/900 (5.2%) without reinfarction (P < 0.001). After the index primary PCI, thrombus was visible in 6/34 (17.7%) with reinfarction when compared with that in 25/952 (2.6%) without reinfarction (P < 0.001), and complete ST-segment resolution in 8/33 (24.2%) compared with that in 462/892 (51.8%, P = 0.002). Mortality at 1 year after the index PCI was higher after reinfarction: 6/34 (17.7%) compared with 53/961 (5.5%, P = 0.003). Patients with stent thrombosis showed myocardial blush grade 0-1 in 11/15 (73.3%) cases, distal embolization in 8/15 (53.3%), and a mortality at 1 year of 4/15 (26.7%). CONCLUSIONS: In contemporary practice with primary PCI and triple antiplatelet therapy for STEMI, the incidence of reinfarction is low. Outcome characteristics after the index PCI were important determinants of reinfarction. However, reinfarction was associated with poor prognosis, and in particular patients with stent thrombosis had poor outcome. PMID- 19309713 TI - Use of a microcatheter in a telescopic system to reach difficult targets in complex congenital heart disease. AB - BACKGROUND: Some lesions can be very difficult to reach, especially if acute angles and/or multiple turns "protect" access. Once reached, the guiding system needs to give sufficient support for balloons or stents to be deployed. METHODS: A "telescopic" system was created consisting of: (1) a microcatheter with guidewire. This system fits into any catheter allowing a 0.035 inch wire. (2) a 4Fr "delivery" catheter. (3) If sharp angles were encountered, the 4Fr catheter was deployed through a 6Fr or larger angulated guiding sheath. This was cut-off 15 cm out of the groin and re-valvulated with a standard short introducer sheath. PATIENTS AND RESULTS: Since 2004, 89 microcatheter telescopic systems were used during a 4-year period in 1,225 procedures (7.3%). The technique allowed probing with a floppy steerable and exchangeable guidewire, securing any gained position as well as exchanges with a stiffer guide wire if required. Procedures where the telescopic system was effective included: complex stenoses e.g. aortopulmonary collaterals (n = 21), tortuous ducts (n = 9), anterograde balloon dilation of critical aortic stenosis (n = 8) and crossing Blalock Taussig shunts (n = 3). Once in place, the telescopic system allowed delivery of embolic material (n = 38), balloon angioplasty (n = 21), stent deployment (n = 23), fulguration of pulmonary valve (n = 1) or introduction of medication (n = 1). The interventionalists felt that using the telescopic system had reduced fluoroscopy and procedure time. CONCLUSIONS: The microcatheter-telescopic system is an invaluable tool to reach difficult targets and allows exchange for suitable guidewires permitting balloons, stents, embolisation material or radiofrequency energy to be deployed in such targets. This approach has become our standard when dealing with difficult targets protected by a tortuous route. PMID- 19309714 TI - Funnel plots for comparing performance of PCI performing hospitals and cardiologists: demonstration of utility using the New York hospital mortality data. AB - BACKGROUND: The New York State Department of Health collects and reports outcome data on the hospitals and cardiologists who perform percutaneous coronary intervention (PCI) to allow them to examine their quality of care. Results are provided in tabular form. However funnel plots are the display method of choice for comparison of institutions and operators, using the principles of statistical process control (SPC). We aimed to demonstrate that funnel plots, which aid a meaningful interpretation of the results, can be derived from the New York PCI dataset. METHODS: The risk-adjusted mortality rates for 48 hospitals and cardiologists performing PCI were used for this analysis. Funnel plots (with control limits at 3 and 2 sigma) of all hospitals and operators performing PCI procedures were generated. Separate plots for emergency and nonemergency PCI procedures were derived. RESULTS: 149,888 patients underwent PCI procedures between January 1, 2002 and December 31, 2004. The 3-year risk-adjusted mortality rates for all PCI patients ranged from 0.00 to 1.37%. The funnel plots show risk adjusted mortality rates against the denominator for that percentage (number of cases), displayed as a scatter plot and compared with the binomial funnel plot calculated around the mean for all cases reported. The risk-adjusted mortality rates of all hospitals were within 3 sigma (99.8%) upper control limits. The risk adjusted mortality rates for three hospitals were above or on the upper warning limit (2 sigma control limit, equating to the 95% confidence interval) and three hospitals had risk-adjusted mortality rates below the 2 and 3 sigma control limits. CONCLUSION: The SPC funnel plot is an easy-to-interpret, risk-adjusted means of identifying units whose performance, in terms of mortality, diverges significantly from the population mean. Funnel plots may be applied to a complex dataset and allow a visual comparison of data derived from multiple healthcare units. Variation is readily identified permitting hospitals and cardiologists to appraise their practices so that effective quality improvement may take place. PMID- 19309715 TI - Paclitaxel-eluting balloon: from bench to bed. AB - Since the first clinical angioplasty by Gruntzig in 1977, restenosis has been the primary drawback of percutaneous coronary intervention (PCI). In the balloon era. restenosis was correlated with elastic recoil and negative remodeling of the arterial wall. Later, introduction of stents proved to be a significant advance in reducing the elastic recoil and negative remodeling at the treatment site but stimulated proliferation, migration of smooth muscle cells, and neointimal hyperplasia, thereby generating a new type of restenosis, in-stent restenosis. Brachytherapy and drug-eluting stents (DES) may be considered the two breakthroughs against neointimal hyperplasia. However, concerns about stent thrombosis and incomplete elimination of in-stent restenosis with DES in complex lesions and patients justify the pursuit of research in this field. Non-stent based local drug delivery and particularly the use of paclitaxel-eluting balloons could be one of these strategies. We aimed to review the concept, preclinical-, and clinical data available with non-stent based local drug delivery and, in particular, with paclitaxel-eluting balloons. PMID- 19309716 TI - Positioning biologic agents in the treatment of Crohn's disease. AB - One decade after the emergence of biologic therapy for Crohn's disease (CD), our treatment algorithms are beginning to change. Once reserved for patients with refractory disease, disease unresponsive to conventional therapies, or those requiring multiple courses of corticosteroids, there is increasing evidence that early, aggressive interventions with immunosuppressants or biologic therapies targeting tumor necrosis factor-alpha or alpha-4 integrins can alter the natural history of CD by reducing the transmural complications of structuring and fistulization and the nearly inevitable requisite for surgical resections. More recent trials are beginning to suggest that intervention with combination therapy for selected patients with a poor prognosis may modify the long-term course of CD. Selection of patients with features predicting a complex or progressive course and early, combined intervention is now possible. Future studies are still needed to best identify predictors of response to individual agents with differing mechanisms of action, as well as to optimize the risk-benefit of long term maintenance therapy. PMID- 19309717 TI - Congenital glioblastoma. AB - We describe the case a 2-day-old female with congenital glioblastoma. Total resection was followed by adjuvant and high dose chemotherapy, as indicated by the current Italian infant protocol. The child is alive and well 18 months after diagnosis. A review of 67 selected congenital brain tumors showed the mortality rate was 82%. Even though the majority of patients had glioblastoma, only 5/67 had received adjuvant therapy. To ensure optimal outcomes, we recommend total or subtotal surgical resection, followed by adjuvant and high dose chemotherapy. Given the lack specific protocols for congenital brain tumors an international consensus seems to be needed, starting with congenital glioblastoma. PMID- 19309718 TI - Negative correlation between cerebrospinal fluid tau protein and cognitive functioning in children with acute lymphoblastic leukemia. AB - The aim of the study was to assess whether cerebrospinal fluid tau protein is associated with cognitive changes in children with acute lymphoblastic leukemia (ALL). Examination of 38 ALL patients revealed a statistically significant increase in tau protein on treatment day 59 and at two points during consolidation phase. Cognitive functioning was examined in 19 patients at an average of 3.7 years after diagnosis. The level of tau at the initiation of maintenance therapy was negatively correlated with verbal abilities measured on an intellectual scale. The study suggests that standard ALL treatment may cause a decline in cognitive functioning. PMID- 19309719 TI - Locating the stem cell niche and tracing hepatocyte lineages in human liver. AB - We have used immunohistochemical and histochemical techniques to identify patches of hepatocytes deficient in the enzyme cytochrome c oxidase, a component of the electron transport chain and encoded by mitochondrial DNA (mtDNA). These patches invariably abutted the portal tracts and expanded laterally as they spread toward the hepatic veins. Here we investigate, using mtDNA mutations as a marker of clonal expansion, the clonality of these patches. Negative hepatocytes were laser capture microdissected and mutations identified by polymerase chain reaction sequencing of the entire mtDNA genome. Patches of cytochrome c oxidase-deficient hepatocytes were clonal, suggesting an origin from a long-lived cell, presumably a stem cell. Immunohistochemical analysis of function and proliferation suggested that these mutations in cytochrome c oxidase-deficient hepatocytes were nonpathogenic. CONCLUSION: These data show, for the first time, that clonal proliferative units exist in the human liver, an origin from a periportal niche is most likely, and that the trajectory of the units is compatible with a migration of cells from the periportal regions to the hepatic veins. PMID- 19309720 TI - Connective tissue growth factor is a Smad2 regulated amplifier of transforming growth factor beta actions in hepatocytes--but without modulating bone morphogenetic protein 7 signaling. AB - In vivo knockdown of connective tissue growth factor (CTGF/CCN2) was recently shown to attenuate the formation of experimental liver fibrosis. The secreted, cysteine-rich growth factor is proposed to adversely modulate the binding of profibrogenic transforming growth factor beta (TGF-beta) and its natural antagonist bone morphogenetic protein (BMP) to their cognate receptors in several cellular systems, but the functionality of CTGF in modulation of the TGF-beta/BMP signaling pathways is still unknown. This study aims at characterizing a potentially differential modulating role of CTGF on TGF-beta- and BMP7-dependent transactivation of reporter gene [Ad-(CAGA)(12)-MLP-luc, Ad-hCTGF-luc, and Ad (BRE)(2)-luc reporter gene] expression in rat hepatocytes. In this context, emphasis is also placed on the differential roles of Smad2 and Smad3 in the TGF beta-dependent transactivation of the endogenous CTGF gene and the CTGF gene reporter, as investigated following adenoviral infection of wild-type and dominant negative Smad2/3 or treatment with the specific inhibitor of Smad3 or ALK5-specific (SB-431542) inhibitor. In this analysis, we found (1) a selective transcriptional activation of the CTGF promoter by Smad2 (but not Smad3); (2) the failure of BMP7 to inhibit the transcriptional activation of the Smad3-selective (CAGA)(12)-luc reporter by TGF-beta, as well as the failure of TGF-beta to inhibit the transcriptional activation of the Smad5-selective (BRE)(2)-luc reporter by BMP7; and (3) the sensitization of hepatocytes toward TGF-beta type I receptor (ALK5)/Smad2 and Smad3-mediated TGF-beta signaling by CTGF, whereas BMP type I receptor (ALK1)/Smad5-mediated BMP7 signaling is not modulated. CONCLUSION: CTGF acts as a Smad2-dependent sensitizer of TGF-beta actions that does not influence BMP7 signaling in hepatocytes. PMID- 19309721 TI - Treatment with immunosuppressives in patients with primary biliary cirrhosis who fail to respond to ursodiol. PMID- 19309722 TI - Expression of a novel marker, Ubc9, in squamous cell carcinoma of the head and neck. AB - BACKGROUND: Ubiquitin-conjugating enzyme (Ubc9) is a novel enzyme involved in posttranslational modification of cellular proteins. The objective of this study was to determine the expression of Ubc9 in squamous cell carcinoma of the head and neck (SCCHN). METHODS: SCCHN specimens were stained with anti-Ubc9 antibodies, scored using a semiquantitative method, and statistically analyzed. RESULTS: Forty-six tumors were stained, 26 of which included adjacent mucosa. Ubc9 was significantly upregulated in the malignant and peritumoral tissues compared with mucosa from normal individuals. In peritumoral tissues, Ubc9 expression was detected in the basal and suprabasal epithelial layers. No Ubc9 was detected in epithelial cells in normal mucosa. These differences in Ubc9 expression were statistically significant (p < .0001). Tumor Ubc9 expression significantly correlated with clinical and pathologic stage. CONCLUSIONS: Ubc9 is significantly overexpressed in the primary SCCHN tumors and peritumoral mucosa compared with normal epithelial cells. These findings suggest that Ubc9 may play an important role in tumorigenesis and tumor progression of SCCHN. PMID- 19309723 TI - Fluorescence in situ hybridization gene amplification analysis of EGFR and HER2 in patients with malignant salivary gland tumors treated with lapatinib. AB - BACKGROUND: Gene amplification status of the epidermal growth factor receptor (EGFR) and the human epidermal growth factor receptor 2 (HER2) were analyzed and correlated with clinical outcome in patients with progressive malignant salivary glands tumors (MSGT) treated with the dual EGFR/Her2 tyrosine kinase inhibitor lapatinib. METHODS: Fluorescence in situ hybridization (FISH) analysis for both EGFR and HER2 gene amplification was performed successfully in the archival tumor specimens of 20 patients with adenoid cystic carcinomas (ACC) and 17 patients with non-ACC, all treated with lapatinib. RESULTS: For ACC, no EGFR or HER2 amplifications were detected. For non-ACC, no EGFR gene amplifications were detected but 3 patients (18%) were HER2 amplified and all had stained 3+ for both EGFR and HER2 by immunohistochemistry (IHC) in their archival specimens. Two of these patients had time-to-progression (TTP) durations of 8.3 months and 18.4 months, respectively. Interestingly, patients with low and high HER2/chromosome specific centromeric enumeration probe (CEP) 17 ratio had a prolonged TTP than those with moderate ratios for both ACC and non-AAC subtypes. CONCLUSIONS: HER2 to CEP17 FISH ratio may predict which patients with MSGT have an increased likelihood to benefit from lapatinib. The finding of HER2:CEP17 ratio as a predictive marker of efficacy to lapatinib warrants further investigation. PMID- 19309724 TI - Radiation technique influence on percutaneous endoscopic gastrostomy tube dependence: Comparison between two radiation schemes. AB - BACKGROUND: Our aim was to determine whether percutaneous endoscopic gastrostomy (PEG) dependence was significantly different between 2 prospective trials with different radiation fractionation schemes. METHODS: Stage III or IV locally advanced head and neck squamous cell carcinomas arising from the oral cavity, hypopharynx, oropharynx, nasopharynx, paranasal sinuses, or larynx were treated using hyperfractionation (A-3 protocol) or accelerated fractionation (A-4 protocol) with chemotherapy. Amifostine was administered 15 to 30 minutes preradiation, at a dose of 500 mg/day in both protocols. It was given as an infusion over 5 to 7 minutes (A-3 protocol) or subcutaneously (A-4 protocol). Data regarding PEG placement and removal were collected prospectively. RESULTS: Thirty-five evaluable A-3 protocol patients, 14 evaluable A-4 protocol patients, and 6 patients treated per A-4 protocol guidelines, but without amifostine as they refused the medication, were included in the analysis. Pretreatment characteristics, such as sex, age, race, T classification, N classification, American Joint Committee on Cancer (AJCC) stage, were compared between the 2 groups of patients. The only significant difference between the 2 groups was AJCC stage. Thirty-five A-3 patients and 20 A-4 patients had overall survivals of 88% versus 80%, 82% versus 75%, and 66% versus 67.5% at 1, 2, and 3 years, respectively (p = .958). With regard to PEG dependence, no significant differences were seen between the 2 groups at 6, 12, or 18 months. CONCLUSION: PEG dependence was not significantly different between the 2 study groups. Type of altered fractionation scheme may not influence PEG dependence in patients treated with similar protocols. Future randomized studies are needed to confirm these findings. PMID- 19309725 TI - Should transition zone biopsies be added to 12-core systematic biopsies of the prostate? AB - PURPOSE: To investigate the value of additional transition zone (TZ) biopsies following 12-core biopsies in the detection and staging of prostate cancer. METHODS: From October 2006 to March 2007, 199 transrectal sonographic (TRUS) guided prostate biopsies (group 1) in 12 peripheral zones (PZ) were performed. Another 199 consecutive patients (group 2) underwent two TZ biopsies in addition to twelve PZ biopsies from March 2007 to July 2007. Mean prostate-specific antigen (PSA) level, prostate volume, Gleason score, and cancer detection rate of each group were compared. The anatomic distribution of prostate cancer in group 2 was also analyzed. RESULTS: Prostate cancer was detected in 76 of 199 patients (38.2%) in group 1 and 71 out of 199 patients (35.3%) in group 2. The cancer detection rate, mean PSA level, prostate volume, and Gleason score were not statistically different in the 2 groups. Cancer was detected by additional TZ biopsies alone in just one out of 71 patients (1.4%). CONCLUSION: Routine TZ biopsies following 12-core systematic biopsies are not warranted for the detection of TZ cancer because of their low additional yield. PMID- 19309726 TI - Association between depression and survival or disease recurrence in patients with head and neck cancer enrolled in a depression prevention trial. AB - BACKGROUND: To determine if depression portends a worse prognosis in patients with head and neck cancer, we conducted a retrospective chart analysis of subjects who participated in a randomized placebo-controlled trial of citalopram for the prevention of depression during head and neck cancer treatment. Thirty five patients were randomized, 34 with 1 or more evaluations form the basis of this report. METHODS: We used the Hamilton Depression Rating Scale (HDRS), with a minimum follow-up of 24 months if no evidence of disease, or until recurrence or death from disease. RESULTS: Fourteen of 34 subjects were depressed at any time during the 16 week RCT (HAMD >15). Seven of the 14 subjects in the depressed group were dead from disease or had recurrence, compared to 4 of 20 who never developed depression (p = .03). Stage of disease was equivalent in the 2 groups. CONCLUSION: This study suggests that depression in patients with head and neck cancer reduces survival. PMID- 19309727 TI - Delta98Delta, a minimalist model of antiparallel beta-sheet proteins based on intestinal fatty acid binding protein. AB - The design of beta-barrels has always been a formidable challenge for de novo protein design. For instance, a persistent problem is posed by the intrinsic tendency to associate given by free edges. From the opposite standpoint provided by the redesign of natural motifs, we believe that the intestinal fatty acid binding protein (IFABP) framework allows room for intervention, giving rise to abridged forms from which lessons on beta-barrel architecture and stability could be learned. In this context, Delta98Delta (encompassing residues 29-126 of IFABP) emerges as a monomeric variant that folds properly, retaining functional activity, despite lacking extensive stretches involved in the closure of the beta barrel. Spectroscopic probes (fluorescence and circular dichroism) support the existence of a form preserving the essential determinants of the parent structure, albeit endowed with enhanced flexibility. Chemical and physical perturbants reveal cooperative unfolding transitions, with evidence of significant population of intermediate species in equilibrium, structurally akin to those transiently observed in IFABP. The recognition by the natural ligand oleic acid exerts a mild stabilizing effect, being of a greater magnitude than that found for IFABP. In summary, Delta98Delta adopts a monomeric state with a compact core and a loose periphery, thus pointing to the nonintuitive notion that the integrity of the beta-barrel can indeed be compromised with no consequence on the ability to attain a native-like and functional fold. PMID- 19309728 TI - Redesign of a protein-peptide interaction: characterization and applications. AB - The design of protein-peptide interactions has a wide array of practical applications and also reveals insight into the basis for molecular recognition. Here, we present the redesign of a tetratricopeptide repeat (TPR) protein scaffold, along with its corresponding peptide ligand. We show that the binding properties of these protein-peptide pairs can be understood, quantitatively, using straightforward chemical considerations. The recognition pairs we have developed are also practically useful for the specific identification of tagged proteins. We demonstrate the facile replacement of these proteins, which we have termed T-Mods (TPR-based recognition module), for antibodies in both detection and purification applications. The new protein-peptide pair has a dissociation constant that is weaker than typical antibody-antigen interactions, yet the recognition pair is highly specific and we have shown that this affinity is sufficient for both Western blotting and affinity purification. Moreover, we demonstrate that this more moderate affinity is actually advantageous for purification applications, because extremely harsh conditions are not required to dissociate the T-Mod-peptide interaction. The results we present are important, not only because they represent a successful application of protein design but also because they help define the properties that should be sought in other scaffolds that are being developed as antibody replacements. PMID- 19309729 TI - Structure of dystrophia myotonica protein kinase. AB - Dystrophia myotonica protein kinase (DMPK) is a serine/threonine kinase composed of a kinase domain and a coiled-coil domain involved in the multimerization. The crystal structure of the kinase domain of DMPK bound to the inhibitor bisindolylmaleimide VIII (BIM-8) revealed a dimeric enzyme associated by a conserved dimerization domain. The affinity of dimerisation suggested that the kinase domain alone is insufficient for dimerisation in vivo and that the coiled coil domains are required for stable dimer formation. The kinase domain is in an active conformation, with a fully-ordered and correctly positioned alphaC helix, and catalytic residues in a conformation competent for catalysis. The conserved hydrophobic motif at the C-terminal extension of the kinase domain is bound to the N-terminal lobe of the kinase domain, despite being unphosphorylated. Differences in the arrangement of the C-terminal extension compared to the closely related Rho-associated kinases include an altered PXXP motif, a different conformation and binding arrangement for the turn motif, and a different location for the conserved NFD motif. The BIM-8 inhibitor occupies the ATP site and has similar binding mode as observed in PDK1. PMID- 19309730 TI - Opposite allosteric mechanisms in TetR and CAP. AB - Regulation of the DNA binding affinity of an oligomeric protein can be considered to consist of an intrinsic component, in which the affinity of an individual DNA binding domain is modulated in response to effector binding, and an extrinsic component, in which the relative position of the protein's two DNA-binding domains are altered so that they can or cannot contact both half-site operators simultaneously. We demonstrated directly that the TetR repressor utilizes an extrinsic mechanism and CAP, the catabolite activator protein, utilizes an intrinsic mechanism. PMID- 19309731 TI - Outcomes after percutaneous coronary intervention of ostial lesions in the era of drug-eluting stents. AB - BACKGROUND: Ostial lesions are a difficult subset associated with suboptimal outcomes after percutaneous coronary intervention (PCI). The aim of this study was to analyze outcomes of ostial lesions in contemporary Australian interventional practice. METHODS: The study population comprised 1,713 consecutive patients who underwent PCI for proximal lesions of the left anterior descending, left circumflex, and right coronary arteries, who were prospectively enrolled in the Melbourne Interventional Group Registry (February 2004-December 2006). We compared the in-hospital, 30-day, and 1-year outcomes of the 109 patients undergoing PCI for ostial, with the 1,604 patients with proximal nonostial lesions. Left main and bifurcation lesions were excluded. RESULTS: Patients in the ostial group were older (mean age 68.8 +/- 11 vs. 64.9 +/- 12 years; P = 0.001), and there was a greater proportion of women (38.5% vs. 28.0%; P = 0.021). Other clinical characteristics were similar. The nonostial group were more likely receive a stent (94.6% vs. 87.2%; P = 0.005) but drug-eluting stents (DES) were deployed more often in the ostial group (47.9% vs. 66.1%; P < 0.0001). There was lower procedural success, with no significant difference in in-hospital death, bleeding or emergency PCI, but unplanned in-hospital coronary artery bypass grafting was more frequent in the ostial group (4.8% vs. 1.0%; P = 0.007). There was no difference in 30-day major adverse cardiac events. However, 12-month death (8.8% vs. 4%, log rank P = 0.032) and MACE (24.2% vs. 13.8%, log rank P = 0.005) were higher in the ostial group than the nonostial group with trends to increased incidence of myocardial infarction (6.6% vs. 4.7%, P = NS), and target vessel revascularization (13.2% vs. 7.9%, P = NS). CONCLUSION: In contemporary, Australian interventional practice, PCI for ostial lesions is associated with a high incidence of adverse outcome at one year despite the introduction of DES. PMID- 19309732 TI - 9,10-Anthraquinone hinders beta-aggregation: how does a small molecule interfere with Abeta-peptide amyloid fibrillation? AB - Amyloid aggregation is linked to a number of neurodegenerative syndromes, the most prevalent one being Alzheimer's disease. In this pathology, the beta-amyloid peptides (Abeta) aggregate into oligomers, protofibrils, and fibrils and eventually into plaques, which constitute the characteristic hallmark of Alzheimer's disease. Several low-molecular-weight compounds able to impair the Abeta aggregation process have been recently discovered; yet, a detailed description of their interactions with oligomers and fibrils is hitherto missing. Here, molecular dynamics simulations are used to investigate the influence of two relatively similar tricyclic, planar compounds, that is, 9, 10-anthraquinone (AQ) and anthracene (AC), on the early phase of the aggregation of the Abeta heptapeptide segment H(14)QKLVFF(20), the hydrophobic stretch that promotes the Abeta self-assembly. The simulations show that AQ interferes with beta-sheet formation more than AC. In particular, AQ intercalates into the beta-sheet because polar interactions between the compound and the peptide backbone destabilize the interstrand hydrogen bonds, thereby favoring disorder. The thioflavin T-binding assay indicates that AQ, but not AC, sensibly reduces the amount of aggregated Abeta(1-40) peptide. Taken together, the in silico and in vitro results provide evidence that structural perturbations by AQ can remarkably affect ordered oligomerization. Moreover, the simulations shed light at the atomic level on the interactions between AQ and Abeta oligomers, providing useful insights for the design of small-molecule inhibitors of aggregation with therapeutic potential in Alzheimer's disease. PMID- 19309734 TI - Neointimal regression-induced incomplete coverage of a bare-metal stent in the left main trunk: serial angiographic and angioscopic evidence obtained by 5-year follow-up. AB - Very late stent thrombosis (VLST), a rare complication of stenting, has been reported to develop more frequently at later than 1 year after the deployment of the drug-eluting stent (DES) compared with the bare-metal stent (BMS). However, the causes for the difference in the incidence remain unknown. Serial angioscopy on 1,591 and 1,952 days after BMS deployment in a Japanese male patient with acute myocardial infarction revealed incomplete neointimal coverage (INC) without the development of cardiovascular event. Therefore, the potential development of VLST remains undeniable in BMS-deployed patients who had INC. PMID- 19309733 TI - Angiographic and clinical outcomes of drug-eluting versus bare metal stent deployment in the Occluded Artery Trial. AB - BACKGROUND: The majority of patients randomized to percutaneous coronary intervention (PCI) in the Occluded Artery Trial (OAT) and its angiographic substudy, the Total Occlusion Study of Canada 2 (TOSCA-2) were treated with bare metal stents (BMS). We aimed to determine if stenting of the target occlusion in OAT with drug-eluting stents (DES) was associated with more favorable angiographic results and clinical outcome when compared with treatment with BMS. METHODS: TOSCA-2 DES was a prospective nonrandomized substudy that provided 1 year angiographic comparison of late loss and reocclusion in 25 patients treated with DES and in 128 treated with BMS. In addition, all PCI-assigned patients enrolled from the time when DES were first utilized were similarly categorized (DES n = 77, and BMS n = 386) and compared using the 3-year cumulative OAT primary combined endpoint of death, myocardial infarction, or Class-IV heart failure, as well as angina. RESULTS: In-segment late loss was 0.14 +/- 0.45 mm for DES and 0.75 +/- 0.86 mm for BMS (P < 0.001). Corresponding binary restenosis rates were 13.0% and 44.3% (P = 0.005). Occlusion at 1 year was observed in 4.0 and 12.1%, respectively (P = 0.23). The 3-year cumulative primary event rate was 13.8% with DES and 12.5% with BMS (hazard ratio 1.08, 99% confidence intervals 0.44, 2.64; P = 0.83). Angina over time occurred less frequently in the DES group (P = 0.01). CONCLUSIONS: Although the reduction of late loss and trend to reduction in reocclusion with the use of DES for PCI of persistently occluded IRA 3-28 days post myocardial infarction did not translate into a signal for reduction in death, reinfarction, or Class IV heart failure, DES use was associated with less angina over time. Further follow-up is warranted. PMID- 19309735 TI - Comparative healing response after sirolimus- and paclitaxel-eluting stent implantation in a pig model of restenosis. AB - OBJECTIVE: We compared local vessel healing and inflammatory responses associated with nonoverlapping sirolimus-eluting stents (SES) and paclitaxel-eluting stents (PES). BACKGROUND: Sirolimus and paclitaxel may have different effects on vascular healing. In the present study, we analyzed the local histologic effects of drug-eluting stents (DES). METHODS: We placed 43 stents (22 PES and 21 SES) in 16 Yucatan minipigs. Stents were randomly assigned and placed in the left anterior descending, circumflex, or right coronary arteries (one stent per artery), covering a region previously injured by balloon angioplasty. RESULTS: Histopathologic analysis showed that the distribution of injury scores was similar between the two stent groups, reflecting the homogeneity of coronary injury secondary to balloon overstretch. Electron microscopy showed complete endothelialization in most cases. Incomplete endothelialization was present in 12.5% of PES and almost 20% of SES at 30 days. In the PES group, moderate to severe inflammation was found in eight arteries, whereas only one vessel had moderate inflammation in the SES group. Severe inflammation was observed significantly more often in the PES than in the sirolimus group (P = 0.006). With the PES group, stent struts overlying side branches had a significantly higher frequency of poor endothelialization scores than did stent struts that did not overlay side branches (P = 0.006). CONCLUSIONS: In this preclinical study in a pig model of in-stent restenosis, implantation of nonoverlapping DES was associated with local inflammatory reactions and decreased endothelial repair. Impaired endothelialization was visualized in the struts overlying side branches. PMID- 19309736 TI - Relevance of weak flavin binding in human D-amino acid oxidase. AB - In the brain, the human flavoprotein D-amino acid oxidase (hDAAO) is involved in the degradation of the gliotransmitter D-serine, an important modulator of NMDA receptor-mediated neurotransmission; an increase in hDAAO activity (that yields a decrease in D-serine concentration) was recently proposed to be among the molecular mechanisms leading to the onset of schizophrenia susceptibility. This human flavoenzyme is a stable homodimer (even in the apoprotein form) that distinguishes from known D-amino acid oxidases because it shows the weakest interaction with the flavin cofactor in the free form. Instead, cofactor binding is significantly tighter in the presence of an active site ligand. In order to understand how hDAAO activity is modulated, we investigated the FAD binding process to the apoprotein moiety and compared the folding and stability properties of the holoenzyme and the apoprotein forms. The apoprotein of hDAAO can be distinguished from the holoenzyme form by the more "open" tertiary structure, higher protein fluorescence, larger exposure of hydrophobic surfaces, and higher sensitivity to proteolysis. Interestingly, the FAD binding only slightly increases the stability of hDAAO to denaturation by urea or temperature. Taken together, these results indicate that the weak cofactor binding is not related to protein (de)stabilization or oligomerization (as instead observed for the homologous enzyme from yeast) but rather should represent a means of modulating the activity of hDAAO. We propose that the absence in vivo of an active site ligand/substrate weakens the cofactor binding, yielding the inactive apoprotein form and thus avoiding excessive D-serine degradation. PMID- 19309737 TI - Abnormal timing of menarche in survivors of central nervous system tumors: A report from the Childhood Cancer Survivor Study. AB - BACKGROUND: Children who receive high-dose radiotherapy to the hypothalamic pituitary (H-P) axis may be at risk for both early and late puberty. To the authors' knowledge, data regarding the risk of altered timing of menarche after higher dose radiotherapy (RT), as used in the treatment of central nervous system (CNS) tumors, are limited. METHODS: The authors evaluated 235 female survivors of CNS tumors, diagnosed between 1970 and 1986, and >1000 sibling controls who were participants in the Childhood Cancer Survivor Study, and provided self-reported data concerning age at menarche. RESULTS: Survivors of CNS tumors were more likely to have onset of menarche before age 10 years compared with their siblings (11.9% vs 1.0%) (odds ratio [OR], 14.1; 95% confidence interval [95% CI], 7.0 30.9). Of the 138 survivors who received RT to the H-P axis, 20 (14.5%) had onset of menarche before age 10 years, compared with 4.3% of those who did not receive RT (OR, 3.8; 95% CI, 1.2-16.5). Age 50 gray OR, 9.0; 95% CI, 2.3-59.5) and spinal RT conferred an increased risk of late menarche, as did older age (>10 years) at the time of diagnosis (OR, 3.0; 95% CI, 1.3-7.0). CONCLUSIONS: Survivors of CNS tumors are at a significantly increased risk of both early and late menarche associated with RT exposure and age at treatment. PMID- 19309739 TI - Application of macroscopic principles to microbial metabolism. PMID- 19309740 TI - CPDadh: a new peptidase family homologous to the cysteine protease domain in bacterial MARTX toxins. AB - A cysteine protease domain (CPD) has been recently discovered in a group of multifunctional, autoprocessing RTX toxins (MARTX) and Clostridium difficile toxins A and B. These CPDs (referred to as CPDmartx) autocleave the toxins to release domains with toxic effects inside host cells. We report identification and computational analysis of CPDadh, a new cysteine peptidase family homologous to CPDmartx. CPDadh and CPDmartx share a Rossmann-like structural core and conserved catalytic residues. In bacteria, domains of the CPDadh family are present at the N-termini of a diverse group of putative cell-cell interaction proteins and at the C-termini of some RHS (recombination hot spot) proteins. In eukaryotes, catalytically inactive members of the CPDadh family are found in cell surface protein NELF (nasal embryonic LHRH factor) and some putative signaling proteins. PMID- 19309741 TI - Application of plant tissue cultures in phytoremediation research: incentives and limitations. AB - The aim of this review is to critically assess the benefits and limitations associated with the use of in vitro plant cell and organ cultures as research tools in phytoremediation studies. Plant tissue cultures such as callus, cell suspensions, and hairy roots are applied frequently in phytoremediation research as model plant systems. In vitro cultures offer a range of experimental advantages in studies aimed at examining the intrinsic metabolic capabilities of plant cells and their capacity for toxicity tolerance. The ability to identify the contributions of plant cells to pollutant uptake and detoxification without interference from microorganisms is of particular significance in the search for fundamental knowledge about plants. However, if the ultimate goal of plant tissue culture experiments is the development of practical phytoremediation technology, the limitations inherent in the use of in vitro cultures as a representative of whole plants in the field must be recognized. The bioavailability of contaminants and the processes of pollutant uptake and metabolite distribution are likely to be substantially different in the two systems; this can lead to qualitative as well as quantitative differences in metabolic profiles and tolerance characteristics. Yet, many studies have demonstrated that plant tissue cultures are an extremely valuable tool in phytoremediation research. The results derived from tissue cultures can be used to predict the responses of plants to environmental contaminants, and to improve the design and thus reduce the cost of subsequent conventional whole plant experiments. PMID- 19309745 TI - Cost implications of new treatments for advanced colorectal cancer. AB - BACKGROUND: Since 1996, 6 new drugs have been introduced for the treatment of metastatic colorectal cancer. Although they are promising, these drugs frequently are given in the palliative and are much more expensive than older treatments. The objective of the current study was to measure the cost implications of treatment with sequential regimens that include chemotherapy and/or monoclonal antibodies. METHODS: A Markov model was used to evaluate a hypothetical cohort of 1000 patients with newly diagnosed, metastatic colorectal cancer. Patients supposedly received up to 3 lines of treatment before supportive care and subsequent death. Data were obtained from published, multicenter phase 2 and randomized phase 3 clinical trials. Sensitivity analyses were conducted on the efficacy, toxicity, and cost. RESULTS: Using drug costs alone, treatment that included new chemotherapeutic agents increased survival at an incremental cost effectiveness ratio (ICER) of $100,000 per discounted life-year (DLY). The addition of monoclonal antibodies improved survival at an ICER of >$170,000 per DLY. The results were most sensitive to changes in the initial regimen. Even with significant improvements in clinical characteristics (efficacy and toxicity), treatment with the most effective regimens still had very high ICERs. CONCLUSIONS: Treatment of metastatic colorectal cancer with the most effective regimens came at very high incremental costs. The authors concluded that cost effectiveness analyses should be a routine component of the drug-development process, so that physicians and patients are informed appropriately regarding the value of new innovations. PMID- 19309746 TI - Lymph node ratio predicts disease-specific survival in melanoma patients. AB - BACKGROUND: The objectives of this analysis were to compare various measures associated with lymph node (LN) dissection and to identify threshold values associated with disease-specific survival (DSS) outcomes in patients with melanoma. METHODS: Patients with lymph node-positive melanoma who underwent therapeutic LN dissection of the neck, axilla, and inguinal region were identified from the SEER database (1988-2005). We performed Cox multivariate analyses to determine the impact of the total number of LNs removed, number of negative LNs removed, and LN ratio on DSS. Multivariate cut-point analyses were conducted for each anatomic region to identify the threshold values associated with the largest improvement in DSS. RESULTS: The LN ratio was significantly associated with DSS for all LN regions. The LN ratio thresholds resulting in the greatest difference in 5-year DSS were .07, .13, and .18 for neck, axillary, and inguinal regions, respectively, corresponding to 15, 8, and 6 LNs removed per positive lymph node. After adjustment for other clinicopathologic factors, the hazard ratios (HRs) were .53 (95% confidence interval [CI], .40 to .71) in the neck, .52 (95% CI, .42 to .65) in the axillary, and .47 (95% CI, .36 to .61) in the inguinal regions for patients who met the LN ratio threshold. CONCLUSIONS: Among the prognostic factors examined, LN ratio was the best indicator of the extent of LN dissection, regardless of anatomic nodal region. These data provide evidence-based guidelines for defining adequate LN dissections in melanoma patients. PMID- 19309747 TI - The effect of modafinil on cognitive function in breast cancer survivors. AB - BACKGROUND: The authors conducted a randomized clinical trial examining the effects of modafinil in reducing persistent fatigue in patients after treatment for cancer and performed secondary analyses to assess the effect of modafinil on cognitive function. METHODS: Breast cancer patients who reported a score of >or=2 on the Brief Fatigue Inventory were enrolled in the study. In phase 1 (P1), patients received 200 mg modafinil open-label once daily for 4 weeks. In phase 2 (P2), patients with a positive response after P1 were randomized either to an additional 4 weeks of modafinil or to placebo. Tests of memory and attention selected from the Cognitive Drug Research (CDR) computerized cognitive assessment were performed at baseline (before modafinil) and after completing phases 1 and 2. The paired differences for each test score were subjected to a Wilcoxon signed rank test. RESULTS: Of the 82 women who were enrolled, 76 completed P1, and 68 completed all assessments in the study. Modafinil had a significant effect on the Speed of Memory (P = .0073) and Quality of Episodic Memory (P < .0001) during P1 of the study. After randomization at Week 8, those patients who continued modafinil demonstrated significantly greater improvement in Speed of Memory (P = .029), Quality of Episodic Memory (P = .0151), and mean Continuity of Attention (P = .0101) relative to the group that was switched to placebo. CONCLUSIONS: The authors found that modafinil improved cognitive performance in breast cancer survivors by enhancing some memory and attention skills. Although confirmation is needed, these findings suggest that modafinil may enhance quality of life in this patient population. PMID- 19309748 TI - Higher expression of chemokine receptor CXCR7 is linked to early and metastatic recurrence in pathological stage I nonsmall cell lung cancer. AB - BACKGROUND: The authors elucidated particular chemokine receptors that are expressed on lung cancer cells, as well as the clinical significance of the expression of these chemokine receptors in completely resected nonsmall cell lung cancer (NSCLC). METHODS: The authors examined gene expression of chemokine receptors (CCR1-11, CXCR1-7, XCR1, and CX3CR1) in 11 cell lines of lung cancer, and gene expression of CXCR3, CXCR4, and CXCR7 (CXCR3/4/7) in surgical specimens of 127 patients who underwent complete resection for their NSCLC between May 2001 and December 2002, using quantitative real-time reverse transcriptase-polymerase chain reaction (PCR). Mutation detection analysis of the EGFR genes using the PCR single-strand conformational polymorphism method was evaluated in patients with pathological (p-) stage I adenocarcinoma. RESULTS: Substantial expression of CXCR3/4/7 mRNA was observed in all NSCLC cell lines examined. In p-stage I NSCLC, CXCR4 and CXCR7 expression values in patients with postoperative metastatic recurrence (Rec-Distant) were significantly higher than in those without recurrences (P = .003 and P = .007, respectively). In addition, the 5-year disease-free survival (DFS) rate of high CXCR7-expressing patients (63.2%) was significantly lower than that of low CXCR7-expressing patients (84.8%) (P = .033). The EGFR mutation was significantly more frequent in patients with higher CXCR7 expression (14 of 21 patients) than in those with lower CXCR7 expression (12 of 32 patients) (P = .038). A multivariate analysis confirmed that high CXCR7 expression was an independent and significant factor predicting a poor DFS in p stage I NSCLC patients (P = .041). CONCLUSIONS: Higher expression of CXCR7 is associated with Rec-Distant and poor DFS in patients with p-stage I NSCLC. PMID- 19309749 TI - Clinical and pathologic prognostic features in acinic cell carcinoma of the parotid gland. AB - BACKGROUND: To the authors' knowledge, the indications for adjuvant treatment in acinic cell carcinoma (AciCC) of the parotid gland have not been elucidated to date. The aim of the current study was to determine patterns of failure and adverse prognostic features. METHODS: Between March of 1989 and August of 2006, 35 patients underwent surgery at Memorial Sloan-Kettering Cancer Center for AciCC of the parotid gland and had their clinical and pathologic features retrospectively analyzed at the primary site. All cases were reviewed by 2 head and neck pathologists. Five-year estimates of survival outcomes were performed, followed by univariate analysis of potential prognostic features. RESULTS: The T classifications were as follows: T1 in 46% of patients, T2 in 23% of patients, T3 in 18% of patients, and T4 in 9% of patients. Three patients had cervical lymph node involvement. All patients underwent surgery as their primary treatment. Approximately 63% of patients (n = 22) received radiation treatment. The median follow-up time for surviving patients was 59.9 months. Five-year estimates of disease-free survival (DFS), overall survival (OS), and local control were 85%, 90%, and 90%, respectively. Of the clinical variables tested, clinical extracapsular extension (ECE), facial nerve sacrifice, and lymph node involvement were found to be significantly associated with a detriment in DFS and OS (P < .05). Positive surgical margins, histologic ECE, >2 mitoses per 10 high-power fields (HPF), atypical mitosis, vascular invasion, perineural invasion, pleomorphism, and necrosis were associated with adverse DFS (P < .05). All of these variables except for vascular invasion (P = .377) and perineural invasion (P = .07) were associated with OS. If high-grade tumors were defined on the basis of high mitotic activity (>2 mitoses/10 HPF) and/or tumor necrosis, high-grade carcinomas had a significantly lower DFS and OS (P = .001). CONCLUSIONS: AciCC had a low treatment failure rate, and a large number of patients could be considered candidates for surgery only. A histologic grading system was devised to help stratify patients for adjuvant treatment. PMID- 19309750 TI - Radical nephrectomy surgical outcomes in the University HealthSystem Consortium Data Base: Impact of hospital case volume, hospital size, and geographic location on 40,000 patients. AB - BACKGROUND: We sought to determine the impact of radical nephrectomy case volume, hospital size, and geographic region on immediate surgical outcomes for patients undergoing radical nephrectomy in academic centers across the country. METHODS: The University HealthSystem Consortium (UHC) Clinical Data Base was queried for data corresponding to patients who underwent radical nephrectomy at 1 of 134 academic medical centers nationwide between 2003 and quarter 2 of 2007 (n = 42,988). Radical nephrectomy case volume (1-99, 100-499, and 500 +), total discharges (1-49,999, 50,000-99,999, 100,000 +), and geographic region (5 categories) were determined for each academic center. ANOVA and the Tukey statistic were used. Length of stay, intensive care unit (ICU) rate, complication (comp) rate, and in-hospital mortality were analyzed. RESULTS: Case volume was a significant predictor of length of stay, ICU, and comp. Mean length of stay was 6.88, 5.61, and 4.76 days, respectively, for centers from lowest to highest case volumes (P < .001). ICU rates for the 3 tiers were 30.77, 17.93, and 12.22 (P < .001). Comp rates were 24.50, 19.40, and 15.48 (P < .001). Tukey analysis revealed a ceiling effect: No differences were seen between the 2 higher case volume groups. Stratification by total discharges revealed differences in ICU rates (P = .001) and comp rates (P = .001). Region of the country had no significant impact on any of the outcome variables in this study. CONCLUSIONS: Radical nephrectomy case volume emerged as an important variable in predicting 3 of the 4 outcome parameters in this study. Results suggest that a "critical volume" of cases portends improved surgical outcomes. PMID- 19309751 TI - Development of a novel HPLC-MS/MS method for the determination of aconitine and its application to in vitro and rat microdialysis samples. AB - A sensitive and selective LC-MS/MS method was developed and validated for the determination of aconitine in microdialysate and rat plasma. Extraction of plasma sample was conducted by use of 1% trichloracetic acid and acetonitrile solution with 10 ng/mL internal standard (propafenone) spiked. Microdialysates were analyzed without sample purification. After sample preparation, 2 microL were injected and separated with an isocratic mobile phase consisting of acetonitrile:0.1% formic acid (60:40, v/v) at a flow rate of 0.3 mL/min. The Agilent G6410A triple quadrupole LC/MS system was operated under the multiple reaction monitoring mode (MRM) using the electrospray ionization technique in positive mode. Overall, the assay exhibited good precision and accuracy. The diffusion properties of aconitine investigated in in vitro microdialysis experiments revealed unfavourable concentration dependence avertable by keeping a constant pH 5.77 using isotonic phosphate buffer solution as perfusate. The mean relative recoveries were 48.23% [coefficient of variation (CV 4.47%)] and 55.38% (CV 2.89%) for retrodialysis and recovery experiments, respectively. The in vivo recovery of aconitine was 34.48% (CV 3.05%) and was stable over the 6 h study period. Following characterization of aconitine both in vitro and in vivo microdialysis, the developed setting is suitable for application in pharmacokinetics and pharmacodynamics studies. PMID- 19309752 TI - Study of influence of additives of tyloxapol on the chromatographic characteristics of the model compounds: the comparative characterization of micellar mobile phases of tyloxapol and Triton X-100. AB - The chromatographic behavior of model compounds of biomedical significance (organic acids, amino acids, drugs) was investigated using mobile phases modified with tyloxapol. The influence of factors such as concentration of tyloxapol, content of organic modifier and pH of mobile phase on the retention factor of solutes was studied. The results were compared with the data obtained by elution with mobile phases containing Triton X-100 additives, since units of Triton X-100 are repeated in the structure of tyloxapol. Divergence in chromatographic behavior of model compounds was explained by the difference in physico-chemical characteristics (microviscosity, polarity, critical micelle concentration, shape of micelles, etc.) of tyloxapol and Triton X-100 micelles. PMID- 19309753 TI - Towards optimum permeability reduction in porous media using biofilm growth simulations. AB - While biological clogging of porous systems can be problematic in numerous processes (e.g., microbial enhanced oil recovery-MEOR), it is targeted during bio barrier formation to control sub-surface pollution plumes in ground water. In this simulation study, constant pressure drop (CPD) and constant volumetric flow rate (CVF) operational modes for nutrient provision for biofilm growth in a porous system are considered with respect to optimum (minimum energy requirement for nutrient provision) permeability reduction for bio-barrier applications. Biofilm growth is simulated using a Lattice-Boltzmann (LB) simulation platform complemented with an individual-based biofilm model (IbM). A biomass detachment technique has been included using a fast marching level set (FMLS) method that models the propagation of the biofilm-liquid interface with a speed proportional to the adjacent velocity shear field. The porous medium permeability reduction is simulated for both operational modes using a range of biofilm strengths. For stronger biofilms, less biomass deposition and energy input are required to reduce the system permeability during CPD operation, whereas CVF is more efficient at reducing the permeability of systems containing weaker biofilms. PMID- 19309754 TI - Directed self-immobilization of alkaline phosphatase on micro-patterned substrates via genetically fused metal-binding peptide. AB - Current biotechnological applications such as biosensors, protein arrays, and microchips require oriented immobilization of enzymes. The characteristics of recognition, self-assembly and ease of genetic manipulation make inorganic binding peptides an ideal molecular tool for site-specific enzyme immobilization. Herein, we demonstrate the utilization of gold binding peptide (GBP1) as a molecular linker genetically fused to alkaline phosphatase (AP) and immobilized on gold substrate. Multiple tandem repeats (n = 5, 6, 7, 9) of gold binding peptide were fused to N-terminus of AP (nGBP1-AP) and the enzymes were expressed in E. coli cells. The binding and enzymatic activities of the bi-functional fusion constructs were analyzed using quartz crystal microbalance spectroscopy and biochemical assays. Among the multiple-repeat constructs, 5GBP1-AP displayed the best bi-functional activity and, therefore, was chosen for self immobilization studies. Adsorption and assembly properties of the fusion enzyme, 5GBP1-AP, were studied via surface plasmon resonance spectroscopy and atomic force microscopy. We demonstrated self-immobilization of the bi-functional enzyme on micro-patterned substrates where genetically linked 5GBP1-AP displayed higher enzymatic activity per area compared to that of AP. Our results demonstrate the promising use of inorganic binding peptides as site-specific molecular linkers for oriented enzyme immobilization with retained activity. Directed assembly of proteins on solids using genetically fused specific inorganic-binding peptides has a potential utility in a wide range of biosensing and bioconversion processes. PMID- 19309755 TI - A DNA replicon system for rapid high-level production of virus-like particles in plants. AB - Recombinant virus-like particles (VLPs) represent a safe and effective vaccine strategy. We previously described a stable transgenic plant system for inexpensive production and oral delivery of VLP vaccines. However, the relatively low-level antigen accumulation and long-time frame to produce transgenic plants are the two major roadblocks in the practical development of plant-based VLP production. In this article, we describe the optimization of geminivirus-derived DNA replicon vectors for rapid, high-yield plant-based production of VLPs. Co delivery of bean yellow dwarf virus (BeYDV)-derived vector and Rep/RepA-supplying vector by agroinfiltration of Nicotiana benthamiana leaves resulted in efficient replicon amplification and robust protein production within 5 days. Co-expression of the P19 protein of tomato bush stunt virus, a gene silencing inhibitor, further enhanced VLP accumulation by stabilizing the mRNA. With this system, hepatitis B core antigen (HBc) and Norwalk virus capsid protein (NVCP) were produced at 0.80 and 0.34 mg/g leaf fresh weight, respectively. Sedimentation analysis and electron microscopy of transiently expressed antigens verified the efficient assembly of VLPs. Furthermore, a single replicon vector containing a built-in Rep/RepA cassette without P19 drove protein expression at similar levels as the three-component system. These results demonstrate the advantages of fast and high-level production of VLP-based vaccines using the BeYDV-derived DNA replicon system for transient expression in plants. PMID- 19309756 TI - Non-viral gene delivery transfection profiles influence neuronal architecture in an in vitro co-culture model. AB - Gene delivery from tissue engineering scaffolds can induce expression of tissue inductive factors to stimulate the cellular processes required for regeneration. Transfected cells secrete diffusible proteins that can create local concentration gradients, depending on the number, distribution, and expression level of transfected cells. These gradients are linked to cellular organization and tissue architecture during embryogenesis. In this report, we investigate neuronal architecture and neurite guidance in response to the concentration gradients achieved by localized secretion of a neurotrophic factor from transfected cells. A co-culture model was employed to examine neuronal responses to multiple transfection profiles, which affects the local concentration of secreted nerve growth factor (NGF). Neuronal architecture, as defined by number of neurites per neuron and length of neurites, was influenced by the transfection profile. Low levels of NGF production by few transfected cells produced longer primary neurites with less branching relative to the higher expression levels or increased numbers of transfected cells. Furthermore, for low NGF production by few transfected cells, the growth cone of the axons was marked by longer extensions and larger surface area, suggesting the presence of a guidance cue. Control studies with varying NGF concentrations did not substantially alter the neuronal architecture, further supporting an effect of localized concentration gradients, and not simply the concentration. Mathematical modeling of NGF diffusion was employed to predict the concentration gradients produced by the transfection profiles, and the resultant gradients were correlated to the cellular response. This report connects the transfection profile, concentration gradients, and the resulting cellular architecture, suggesting a critical design consideration for the application of gene delivery to regenerative medicine. PMID- 19309757 TI - High-performance liquid chromatography method for determination of carnosic acid in rat plasma and its application to pharmacokinetic study. AB - A sensitive and reproducible high-performance liquid chromatography method with ultraviolet detection (UV) was developed for the determination of carnosic acid (CA) in rat plasma. After simple acidification and liquid-liquid extraction of plasma samples using gemfibrozil as an internal standard, the supernatant was evaporated to dryness under a gentle stream of nitrogen. The residue was reconstituted in 200 microL before being injected into the chromatographic system. The analysis was performed on a C(18) column protected by an ODS guard column using acetonitrile-0.1% phosphoric acid (55:45, v/v) as mobile phase, and the wavelength of the UV detector was set at 210 nm. The calibration curve was linear over the range of 0.265-265.0 microg/mL with a correlation coefficient of 0.9997. The recovery for plasma samples of 0.530, 13.25, 132.5 and 265.0 microg/mL was 72.2, 87.9, 90.4 and 94.7%, respectively. The intra-day and inter day relative standard deviations for the measurements of quality control samples were less than 3.1%. The stability of the plasma samples was also validated. This method was successfully used to study the pharmacokinetics and bioavailability of CA in rats. PMID- 19309758 TI - The role of the dorsal raphe nucleus in the development, expression, and treatment of L-dopa-induced dyskinesia in hemiparkinsonian rats. AB - Convergent evidence indicates that in later stages of Parkinson's disease raphestriatal serotonin neurons compensate for the loss of nigrostriatal dopamine neurons by converting and releasing dopamine derived from exogenous administration of the pharmacotherapeutic L-3,4-dihydroxyphenyl-L-alanine (L dopa). Because the serotonin system is not equipped with dopamine autoregulatory mechanisms, it has been postulated that raphe-mediated striatal dopamine release may fluctuate dramatically. These fluctuations may portend the development of abnormal involuntary movements called L-dopa-induced dyskinesia (LID). As such, it has been hypothesized that reducing the activity of raphestriatal neurons could dampen supraphysiological stimulation of striatal dopamine receptors thereby alleviating LID. To directly address this, the current study employed the rodent model of LID to investigate the contribution of the rostral raphe nuclei (RRN) in the development, expression and treatment of LID. In the first study, dual serotonin/dopamine selective lesions of the RRN and medial forebrain bundle, respectively, verified that the RRN are essential for the development of LID. In a direct investigation into the neuroanatomical specificity of these effects, microinfusions of +/-8-OH-DPAT into the intact dorsal raphe nucleus dose dependently attenuated the expression of LID without affecting the antiparkinsonian efficacy of L-dopa. These current findings reveal the integral contribution of the RRN in the development and expression of LID and implicate a prominent role for dorsal raphe 5-HT1AR in the efficacious properties of 5-HT1AR agonists. PMID- 19309759 TI - Arrestin3 mediates D(2) dopamine receptor internalization. PMID- 19309760 TI - Treatment of hypersexuality in Parkinson's disease with carbamazepine--a case report. PMID- 19309761 TI - Lysophosphatidylcholine--biomarker of Metformin action: studied using UPLC/MS/MS. AB - An UPLC/MS/MS based metabonomic method was developed and applied to the elucidation of biomarker of metformin action. The plasma metabolite profiling in healthy volunteers before and after per os metformin was determined with UPLC/MS/MS and analyzed by using multivariate statistics. Significant difference in endogenous metabolite profiles was revealed before and after administration of metformin. Four biomarkers found were lysophosphatidylcholines (LPCs), and their structures were tentatively identified to be 16:0 LPC, 18:0 LPC, 18:1 LPC and 18:2 LPC according to the molecular ions information and corresponding fragments of product ion scan. Lysophosphatidylcholine in blood may be involved in metformin treatment. PMID- 19309762 TI - Bioanalytical methods for the determination of itraconazole and hydroxyitraconazole: overview from clinical pharmacology, pharmacokinetic, pharmacodynamic and metabolism perspectives. AB - Itraconazole represents an important therapeutic option for the treatment of fungal infections. Itraconazole undergoes rapid metabolism to form hydroxyitraconazole, which also contributes to the anti-fungal activity exhibited by the parent compound. Since both itraconazole and hydroxyitraconazole are effective inhibitors of cytochrome P450 (CYP) 3A4 and p-glycoprotein (pgp) mediated efflux transporters, they have the potential to elicit drug-drug interaction with a number of CYP3A4 and/or pgp substrates. This review focuses on providing comprehensive details on the bioanalytical methods available for the quantitation of both itraconazole and hydroxyitraconazole. Additionally, it provides an overview of the clinical pharmacology (several case studies of drug drug interactions), pharmacokinetics, pharmacodynamics and metabolism related aspects of itraconazole. PMID- 19309763 TI - Reversed-phase liquid chromatographic determination of enantiomers of atenolol in rat plasma using derivatization with Marfey's reagent. AB - An HPLC method was established for enantioseparation of (R,S)-atenolol (ATE) and determination of enantiomers in rat plasma. Marfey's reagent (1-fluoro-2,4 dinitrophenyl-5-L-alanine amide, FDNP-L-Ala-NH(2), MR) was used as chiral derivatizing reagent with detection of diastereomers at 340 nm. It was shown that the R-isomer eluted before the S-isomer. The method was validated for linearity, repeatability, limits of detection and limit of quantification (LOQ). Recovery of ATE at LOQ was 92.8% for (R)-ATE and 92.6% for (S)-ATE. PMID- 19309764 TI - Simultaneous determination of six herbal components in intestinal perfusate by high-performance liquid chromatography. AB - An effective, accurate and reliable HPLC with UV detection method was developed and validated for quantitation of six components: baicalin, berberine hydrochloride, quercetin, kaempferol, isorhamnetin and baicalein in intestinal perfusate using rotundin as an internal standard. The chromatographic separation was performed on a Welchrom-C(18) column (250 x 4.6 mm i.d. with 5.0 microm particle size) with a mobile phase consisting of acetonitrile, water, phosphoric acid and triethylamine (30:70:0.2:0.1,v/v) at a flow rate of 1.0 mL/min and a UV detection at 270 nm. The method had a chromatographic run time of 30 min and excellent linear behavior over the investigated concentration ranges observed with the values of r higher than 0.99 for all the analytes. The lower limit of quantification of the analytical method was 0.09 microg/mL for berberine hydrochloride, quercetin, kaempferol and baicalein and 0.18 microg/mL for baicalin and isorhamnetin. The intra- and inter-day precisions measured at three concentration levels were all less than 10% for all analytes. The bias ranged from -6.91 to 4.33%. The validated method has been successfully applied to investigate the rat intestine absorption profiles of baicalin, berberine hydrochloride, quercetin, kaempferol, isorhamnetin and baicalein. PMID- 19309765 TI - Quantification of montelukast, a selective cysteinyl leukotriene receptor (CysLT1) antagonist in human plasma by liquid chromatography-mass spectrometry: validation and its application to a human pharmacokinetic study. AB - A highly sensitive, rapid assay method has been developed and validated for the estimation of montelukast (MTK) in human plasma with liquid chromatography coupled to tandem mass spectrometry with electro spray ionization in the positive ion mode. Liquid-liquid extraction was used to extract MTK and amlodipine (internal standard, IS) from human plasma. Chromatographic separation was achieved with 10 mM ammonium acetate (pH 6.4): acetonitrile (15:85, v/v) at a flow rate of 0.50 mL/min on a Discovery HS C(18) column with a total run time of 3.5 min. The MS/MS ion transitions monitored were 586.10 --> 422.10 for MTK and 409.20 --> 238.30 for IS. Method validation and clinical sample analysis were performed as per FDA guidelines and the results met the acceptance criteria. The lower limit of quantitation achieved was 0.25 ng/mL and linearity was observed from 0.25 to 800 ng/mL. The intra-day and inter-day precisions were 5.97-8.33 and 7.09-10.13%, respectively. This novel method has been applied to a pharmacokinetic study of MTK in humans. PMID- 19309766 TI - The effect of age on osteogenic, adipogenic and proliferative potential of female adipose-derived stem cells. AB - Human adipose tissue is an ideal source of autologous cells that is both plentiful and easily obtainable in large quantities through the simple surgical procedure of liposuction. The stromal vascular fraction of adipose tissue contains a stem cell population, adipose-derived stem cells (ASCs), capable of adipogenic, osteogenic, myogenic and chondrogenic differentiation. These cells have already been recognized to possess great therapeutic potential in tissue engineering and regeneration. In this study, we sought to determine the effect of donor age on the growth kinetics and differentiation potential of ASCs. For this, ASCs were isolated from liposuctioned adipose tissue obtained from female patients in the age range 20-58 years. Population doubling time was calculated over 2 weeks and differentiation potential was determined by assaying for adipogenesis and osteogenesis. ASCs obtained from older donors appeared to have a slower rate of proliferation, but this relationship was not significant. While adipogenic potential was unrelated to donor age, a distinct relationship between donor age and osteogenic potential was observed. The aetiology of this age dependent change in osteogenic potential was not due to any changes in the number of precursors with osteogenic capacity in the adipose sample. These findings have important implications for emerging cell-based therapeutic strategies, such as tissue engineering, in addition to treatment of various metabolic bone disorders including osteoporosis. PMID- 19309767 TI - Long-term care: funding and insurance. End-of-year issue brief. PMID- 19309768 TI - Ginkgo might not be a brain saver. PMID- 19309769 TI - The curious case of the ageing cells. PMID- 19309770 TI - Spontaneous progression of ligatured induced peri-implantitis at implants with different surface characteristics. An experimental study in dogs II: histological observations. AB - AIM: The aim of the present study was to analyze tissue reactions to plaque formation following ligature removal in experimental peri-implantitis at commercially available implants in dogs. MATERIAL AND METHODS: Mandibular premolars and the three anterior premolars in both sides of the maxilla were extracted in six Labrador dogs. After 3 months, four implants representing four different implant systems--groups A (turned), B (TiOblast), C (SLA), D (TiUnite)- were placed in a randomized order in the right side of the mandible. Three months after implant installation, experimental peri-implantitis was initiated by placement of ligatures and plaque formation. The ligatures were removed when about 40-50% of the supporting bone was lost. After the subsequent 24-week period of continued plaque accumulation, block biopsies containing implants and their surrounding tissues were obtained and prepared for histological analysis. RESULTS: All types of implants exhibited extensive inflammatory cell infiltrates and large associated crater-formed osseous defects. The lesions were consistently characterized by insufficient encapsulation of pus and biofilm layers and the inflammatory cell infiltrates extended apical of the pocket epithelium. The presence of numerous osteoclasts indicated active tissue destruction. The vertical dimension and the overall surface area of the infiltrated connective tissue (ICT) were larger at implants of group D than at other implant types. CONCLUSION: It is suggested that spontaneous progression of peri-implantitis is associated with severe inflammation and tissue destruction. PMID- 19309771 TI - Bone formation with various bone graft substitutes in critical-sized rat calvarial defect. AB - OBJECTIVE: This histomorphometric study compared the efficacy of a new bone graft substitute (N-HA) derived from hen eggshell, consisted of submicron scale porous hydroxyapatite structure, in the healing of 8 mm diameter critical size defects in rat calvaria. We compared N-HA alone or in combination with calcium sulfate (CS), with a commercial bone substitute, anorganic bovine bone (Bio-Oss, BO). MATERIAL AND METHODS: Critical size defects were created in calvaria of 56 adult Sprague-Dawley rats. Animals were divided into four groups and treated with (1) unfilled defects, (2) N-HA grafts, (3) BO grafts and (4) N-HA/CS grafts. The percentage of new bone formed (NB%) was evaluated histomorphometrically after 6 and 12 weeks. RESULTS: The N-HA group exhibited more new bone formation compared with other groups at 6 and 12 weeks. Histomorphometric analysis showed greater NB% in N-HA group (11.2% at 4 weeks and 19.2% at 12 weeks) compared with those in unfilled (3.9% at 6 weeks and 6.4% at 12 weeks), BO-treated (6.4% at 6 weeks and 8.2% at 12 weeks) and N-HA/CS-treated (6.3% at 6 weeks and 12.6% at 12 weeks) groups. The N-HA group showed significant differences in NB% compared with unfilled group at 6 weeks (P=0.016), unfilled and BO-treated groups at 12 weeks (P=0.001). The addition of CS did not enhance the NB% compared with defects grafted with N-HA alone. CONCLUSION: N-HA was an osteoconductive bone substitute for treating osseous defects in critical size defects of rat calvaria. PMID- 19309772 TI - Effect of cell density on adipogenic differentiation of mesenchymal stem cells. AB - The effect of cell density on the adipogenic differentiation of human bone marrow derived mesenchymal stem cells (MSCs) was investigated by using a patterning technique to induce the formation of a cell density gradient on a micropatterned surface. The adipogenic differentiation of MSCs at a density gradient from 5 x 10(3) to 3 x 10(4) cells/cm2 was examined. Lipid vacuoles were observed at all cell densities after 1-3 weeks of culture in adipogenic differentiation medium although the lipid vacuoles were scarce at the low cell density and abundant at the high cell density. Real-time RT-PCR analysis showed that adipogenesis marker genes encoding peroxisome proliferator-activated receptor gamma2 (PPARgamma2), lipoprotein lipase (LPL), and fatty acid binding protein-4 (FABP4) were detected in the MSCs cultured at all cell densities. The results suggest that there was no apparent effect of cell density on the adipogenic differentiation of human MSCs. PMID- 19309773 TI - "Twin" viruses. PMID- 19309774 TI - Increased ribosomal biogenesis induces pancreatic beta cell failure in mice model of type 2 diabetes. AB - AIM: To study the changes in gene expression by pancreatic beta cells under insulin resistance conditions. METHOD: An exhaustive gene expression analysis was performed, using isolated pancreatic islets of obese diabetic model Lepr(-/-) mice. Overexpression of cyclin D2 was induced in cells from the pancreatic beta cell line, namely, INS-1. RESULTS: Through a gene expression analysis using islets isolated from db/db mice, we found a significant increase in the expression of ribosome-related molecules. In addition, increased expression of cyclin D2 was found at certain protein levels. As INS-1 cells were induced to overexpress cyclin D2, we found an increase in the expression of ribosome-related molecules. Concurrently, an increase in the expression of endoplasmic reticulum stress (ER stress)-related molecules was also found. CONCLUSION: In cases of pancreatic beta cell hyperplasia associated with insulin resistance, ribosomal biogenesis is increased, and ER stress is induced. PMID- 19309775 TI - Intramucosal distribution of WNT signaling components in human esophagus. AB - BACKGROUND: The molecular mechanisms governing the biology and pathobiology of esophageal squamous mucosa in health and disease are not completely understood. Earlier genome-wide expression study of normal-looking esophageal squamous mucosa has shown differential expression of the Wingless-type MMTV integration site family (Wnt) modulators Dickkopf (Dkk) homologs among healthy individuals and patients with reflux esophagitis and Barrett metaplasia suggesting that the Wnt pathway may be involved in esophageal mucosal biology. STUDY: Seven full thickness human donor esophagi were cryosectioned for immunohistochemical analysis, and lamina propria (LP), basal (BC), intermediate (IC), and superficial (SC) cells were also dissected by laser-capture microdissection for real-time polymerase chain reaction. RESULTS: Wnt1, 2b, and 3a were expressed primarily in BC, Wnt3, and 5b in LP, and Wnt5a in IC. Frizzled 1, low-density lipoprotein receptor-related protein 6, secreted frizzled-related protein 1, T-cell-specific transcription factor 3, and dishevelled 3 were expressed highest in LP decreasing precipitously medially toward SC. Dkk1 predominantly expressed in SC was more than 100-folds greater than other layers (P<0.001). Dkk4 was expressed primarily in SC but Dkk3 was opposite with greatest expression in LP. Immunohistochemical analysis showed Wnt1 and 3a in BC, Wnt5a in IC and SC, Dkk1 predominantly in SC, Dkk4 in SC and IC, and Dkk3 and SFRP1 in LP and BC PMID- 19309776 TI - Novel human pathological mutations. Gene symbol: SLC34A2. Disease: pulmonary alveolar microlithiasis. PMID- 19309777 TI - Hypereosinophilia due to mesalazine treatment. PMID- 19309778 TI - Novel human pathological mutations. Gene symbol: NPHS1. Disease: congenital nephrotic syndrome, Finnish type. PMID- 19309779 TI - Novel human pathological mutations. Gene symbol: PYGM. Disease: McArdle disease. PMID- 19309780 TI - Novel human pathological mutations. Gene symbol: HSD17B3. Disease: 17 beta hydroxysteroid dehydrogenase-3 deficiency. PMID- 19309781 TI - Novel human pathological mutations. Gene symbol: GLA. Disease: Fabry disease. PMID- 19309782 TI - Novel human pathological mutations. Gene symbol: APC. Disease: adenomatous polyposis coli. PMID- 19309783 TI - Novel human pathological mutations. Gene symbol: PYGM. Disease: McArdle disease. PMID- 19309784 TI - Novel human pathological mutations. Gene symbol: SLC40A1. Disease: haemochromatosis, type 4. PMID- 19309785 TI - Novel human pathological mutations. Gene symbol: PHEX. Disease: rickets, hypophosphataemic. PMID- 19309786 TI - Novel human pathological mutations. Gene symbol: PYGM. Disease: McArdle disease. PMID- 19309787 TI - Novel human pathological mutations. Gene symbol: PKLR. Disease: pyruvate kinase deficiency. PMID- 19309788 TI - Novel human pathological mutations. Gene symbol: AR. Disease: androgen insensitivity syndrome. PMID- 19309789 TI - Disappointed with the details. PMID- 19309790 TI - Retraction notice to "Development and validation of an LC-MS method with electrospray ionization for quantitation of digoxin in human plasma and urine: application to a pharmacokinetic study" [J. Chromatogr. B 875 (2008) 405-410]. PMID- 19309791 TI - Chemical and physical stimuli induce cardiomyocyte differentiation from stem cells. AB - In this study we investigated cardiomyocyte differentiation of rat bone marrow mesenchymal stem cells (BM-MSCs) by treating the stem cells with conditions mimicking that of myocardial infarction. The extract from infarcted rat myocardium contained the biochemical factors arising after infarction. The cardiac contraction and relaxation were simulated by applying 4% strain at 1 Hz to the stem cells. We found that the extract from infarcted myocardium or 4% strain each alone could induce cardiomyocyte differentiation of BM-MSCs, as shown by expression of cardiomyocyte-specific genes including alpha-actin, connexin 43, Nkx2.5, MEF2c, GATA4, alpha-MHC, and Troponin I. Furthermore, a combination of the extract and 4% strain had stronger effects on cardiomyocyte differentiation than what either treatment alone had. Our results suggest that this in vitro model system simulates the local cardiac environment cues after infarction and may be useful in identifying the biochemical and physical factors involved in cardiomyocyte differentiation. PMID- 19309792 TI - The role of the Ser/Thr cluster in the phosphorylation of PPPSP motifs in Wnt coreceptors. AB - Wnt/beta-catenin signaling controls a variety of cellular processes, including cell growth, oncogenesis, and development. Upon Wnt stimulation, the intracellular region of the coreceptor, LRP6 or 5, is phosphorylated by the membrane-recruited GSK3beta and CK1. The cytoplasmic domain of LRP6/5 contains one Ser/Thr cluster and the PPPSP motifs, both of which are essential for propagation of the signal. While the phosphorylated PPPSP motifs are known to directly inhibit GSK3beta, the biochemical role of the phosphorylated Ser/Thr cluster remains to be elucidated. Herein, we reveal that the Ser/Thr cluster plays an important role in the phosphorylation of the PPPSP motif. Interestingly, we observe that GSK3beta activity on the PPPSP motif requires a high ATP concentration, close to that of the physiological condition. Taken together, these data suggest that the phosphorylated Ser/Thr cluster serves as a docking site for GSK3beta to promote the phosphorylation of the PPPSP motif. Our results provide insight into the molecular mechanism for the initial events of the Wnt/beta-catenin signaling. PMID- 19309793 TI - Controlling data flow enhances anesthesiology's role in perioperative care. PMID- 19309794 TI - Hepatitis C virus infection. PMID- 19309795 TI - Novel human pathological mutations. Gene symbol: ALPL. Disease: hypophosphatasia. PMID- 19309796 TI - Novel human pathological mutations. Gene symbol: WNT7A. Disease: ulnar and fibula absence, with severe limb deficiency. PMID- 19309797 TI - Rethinking a drug treatment failure on a traditional ALS target. AB - In a recent issue of Experimental Neurology, Boston-Howes and colleagues used an assay of glutamate transport to screen 1040 FDA approved drugs in an attempt to identify compounds that would increase glutamate transport, a central function of astrocytes, and a potential biological target for neuroprotection for a variety of neurological disorders. They identified the compound nordihydroguaiaretic acid (NDGA) as a particularly good candidate for inducing glutamate transport. Pharmacological increases in glutamate transport could have a number of potential applications to diseases of the nervous system where glutamate excitotoxicity is thought to be a contributing factor to pathogenesis including Amyotrophic Lateral Sclerosis, Alzheimer's disease, Parkinson's disease, stroke, and epilepsy among others. They chose to test this compound in a model of Amyotrophic Lateral Sclerosis (ALS)--the SOD1G93A mouse. In both human ALS and rodent models of the disease, glutamate excitotoxicity and abnormalities in glutamate transporter biology more specifically, have been implicated in ALS disease propagation. Interestingly, while the authors nicely demonstrate that NDGA has a biological effect on glutamate transport in normal (wild type) central nervous system tissues both in vitro and in vivo, it was the somewhat unexpected (and often overlooked) findings in the ALS mouse model that makes this manuscript notable and suggests that rethinking translational approaches to drug therapies in ALS may be on the horizon. PMID- 19309798 TI - Novel human pathological mutations. Gene symbol: NROB1. Disease: adrenal hypoplasia. PMID- 19309799 TI - Novel human pathological mutations. Gene symbol: CPS1. Disease: carbamoyl phosphate synthetase I deficiency. PMID- 19309800 TI - Novel human pathological mutations. Gene symbol: GLA. Disease: Fabry disease. PMID- 19309801 TI - Novel human pathological mutations. Gene symbol: CDH1. Disease: gastric cancer. PMID- 19309802 TI - Novel human pathological mutations. Gene symbol: PYGM. Disease: McArdle disease. PMID- 19309803 TI - Novel human pathological mutations. Gene symbol: FBN1. Disease: Marfan syndrome. PMID- 19309804 TI - Novel human pathological mutations. Gene symbol: PYGM. Disease: McArdle disease. PMID- 19309805 TI - Novel human pathological mutations. Gene symbol: PKLR. Disease: pyruvate kinase deficiency. PMID- 19309806 TI - Novel human pathological mutations. Gene symbol: OCA2. Disease: albinism, oculocutaneous II. PMID- 19309808 TI - Gangrene: Lecture delivered at the Royal College of Surgeons of England on 10th April, 1947. PMID- 19309807 TI - Foreword. PMID- 19309809 TI - A Hundred Years of Gastric Surgery: An expanded version of an address delivered at the Centenary Meeting of the American Medical Association at Atlantic City on 11th June 1947. PMID- 19309810 TI - The Library During the War. PMID- 19309812 TI - Anaesthesia Centenary. PMID- 19309811 TI - The Museum of Pathology. PMID- 19309813 TI - Vicary Lecture for 1945: The Renaissance and its Influence on English Medicine Surgery and Public Health. PMID- 19309815 TI - Richard Owen. PMID- 19309814 TI - Surgery in Two Wars: 1914-18, 1939-45. PMID- 19309816 TI - Painful Tumours of the Skin: Post-Graduate Lecture given at The Royal College of Surgeons of England on 28th February, 1947. PMID- 19309817 TI - The Surgical Aspects of Intestinal Amoebiasis: Hunterian Lecture delivered at The Royal College of Surgeons of England on 5th June, 1947. PMID- 19309818 TI - Recent Advances in the Treatment of Carcinoma of the Oesophagus: Surgery Lecture delivered at The Royal College of Surgeons of England on 11th April, 1947. PMID- 19309819 TI - Injuries and Cicatricial Stenosis of the Larynx: Post-Graduate Lecture delivered at The Royal College of Surgeons of England on 7th May, 1947. PMID- 19309820 TI - Vicary Lecture For 1946: Naval Medicine in the Ages of Elizabeth and James. PMID- 19309821 TI - Blood Transfusion and Intravenous Therapy: Post-Graduate Lecture delivered at The Royal College of Surgeons of England on 24th April, 1947. PMID- 19309822 TI - The Replacement of Skin Losses in Traumatic Injuries: Surgery Lecture delivered at The Royal College of Surgeons of England on 21st April, 1947. PMID- 19309824 TI - The Ceremonial Mace of The Royal College of Surgeons. PMID- 19309823 TI - Foods and Feeding as they Affect Teeth and Their Environment: Charles Tomes Lecture delivered at The Royal College of Surgeons of England on 24th July, 1947. PMID- 19309825 TI - Report of Contributions by Fellows of The American College of Surgeons. PMID- 19309826 TI - Physiological Responses To Wounding: Moynihan Lecture delivered at The Royal College of Surgeons of England on 8th July, 1947. PMID- 19309827 TI - Post-Operative Pulmonary Embolism: Lecture delivered at The Royal College of Surgeons of England on 24th July, 1947. PMID- 19309828 TI - The Use of D-Tubocurarine Chloride in Anaesthesia: Lecture delivered at The Royal College of Surgeons of England on 17th April, 1947. PMID- 19309829 TI - Operative Procedures in Polio-Myelitis: Lecture delivered at The Royal College of Surgeons of England on 30th May, 1947. PMID- 19309830 TI - Hunter's Bust in Leicester Square. PMID- 19309831 TI - Dinner to Sir Alfred and Lady Webb-Johnson. PMID- 19309832 TI - Robin Adair. PMID- 19309833 TI - Lister's Last Assistant. PMID- 19309834 TI - W. E. Henley and Lister. PMID- 19309835 TI - "Observables" at the Royal College of Surgeons: 1. Woolner's "Puck". PMID- 19309836 TI - Sir William H. Collins. PMID- 19309837 TI - Faculty of Dental Surgery. PMID- 19309839 TI - Vicary Lecture for 1947: Lawson Tait. PMID- 19309838 TI - "Observables" at the Royal College of Surgeons: 2. The Ranby Cup. PMID- 19309840 TI - "Observables" at the Royal College of Surgeons: 4. The Cheselden Cup. PMID- 19309841 TI - "Observables" at the Royal College of Surgeons: 5. John Hunter's Tankard. PMID- 19309843 TI - "Observables" at the Royal College of Surgeons: 7. The Erasmus Wilson Tankard and Goblets. PMID- 19309842 TI - "Observables" at the Royal College of Surgeons: Truffot's Statue of Jupille, The Shepherd Boy. PMID- 19309845 TI - "Observables" at the Royal College of Surgeons: 9. Gilbert's Bust of sir Richard Owen. PMID- 19309844 TI - "Observables" at the Royal College of Surgeons: 8. The Fellows' Cups. PMID- 19309846 TI - "Observables" at the Royal College of Surgeons: 10. "Mors Janua Vitae". PMID- 19309848 TI - "Observables" at the Royal College of Surgeons: 12. A Crimean Casualty. PMID- 19309847 TI - "Observables" at the Royal College of Surgeons: 11. Sir Jonathan Hutchinson and other Members of the Court of Examiners. PMID- 19309850 TI - Sibert Forrest Cowell. PMID- 19309849 TI - Observables at the Royal College of Surgeons: (13) A Bronze Ecorche Statuette. PMID- 19309851 TI - "Observables" at the Royal College of Surgeons: 14. The Rowlandson Water-Colours. PMID- 19309852 TI - The Hunterian Oration for 1949: Delivered at the Royal College of Surgeons of England on 14th February, 1949. PMID- 19309853 TI - John Hunter's Account of a "Mortifyd" Appendix Caeci. PMID- 19309855 TI - "Observables" at the Royal College of Surgeons: 15. Pictures by George Stubbs, R.A. PMID- 19309854 TI - Wilfred Trotter: Presidential Address given at the meeting of the Surgical Section of The Royal Society of Medicine on 3rd November, 1948. PMID- 19309856 TI - The Faculty of Anaesthetists. PMID- 19309857 TI - "Observables" at the Royal College of Surgeons: 16. The Arms of the Company and College. PMID- 19309858 TI - The Life and Achievements of Joseph Thomas Clover: The Joseph Clover Lecture delivered at the Royal College of Surgeons on 16th March, 1949. PMID- 19309859 TI - "Observables" at the Royal College of Surgeons: 17. Records of Admiral Horatio Nelson's Wounds. PMID- 19309860 TI - Opening of Residential Quarters: Royal College of Surgeons of England. PMID- 19309862 TI - The Honorary Medal of the College. PMID- 19309861 TI - John Hunter and his Museum: An Address delivered to the Osler Club on 10th December, 1948. PMID- 19309863 TI - "Observables" at the Royal College of Surgeons: 18. The Jacksonian Prize. PMID- 19309864 TI - Unpublished Letters of William Clift. PMID- 19309865 TI - The Joseph Henry Lectureship. PMID- 19309867 TI - "Observables" at the Royal College of Surgeons: 19. The Blane Medal. PMID- 19309866 TI - Conferring of the Honorary Fellowship on Viscount Nuffield. PMID- 19309869 TI - "Observables" at the Royal College of Surgeons: 20. The Plate of The Faculty of Dental Surgery. PMID- 19309868 TI - A. H. G. Doran. PMID- 19309870 TI - Faculty of Dental Surgery: Second Anniversary, July 15th, 1949. PMID- 19309872 TI - A Patient Remembers Lister. PMID- 19309871 TI - Recollections of Lord Lister. PMID- 19309873 TI - "Observables" at the Royal College of Surgeons: 21. A Silver Shaving Set. PMID- 19309874 TI - The Portraiture of William Harvey. PMID- 19309875 TI - Cardio-Vascular Anomalies. PMID- 19309876 TI - "Observables" at the Royal College of Surgeons: 22. Frere Jacques. PMID- 19309877 TI - Gift for Residential College. PMID- 19309878 TI - "Observables" at the Royal College of Surgeons: 23. The Fellowship Signature Book. PMID- 19309879 TI - "Observables" at the Royal College of Surgeons: 24. Clocks. PMID- 19309880 TI - Appointment by Army Council. PMID- 19309881 TI - "Observables" at the Royal College of Surgeons: 25. The President's Badge Of Office. PMID- 19309883 TI - "Observables" at the Royal College of Surgeons: 26. Lister's "Commonplace Books" Observations on Penicillium. PMID- 19309882 TI - Inflammation and Repair: Lectures delivered at the Royal College of Surgeons of England on 10th February, 1949. PMID- 19309884 TI - Scoliosis: Lecture delivered at the Royal College of Surgeons of England on 7th October, 1949. PMID- 19309885 TI - The Development of the Mammary Gland: Arris and Gale Lecture, delivered at the Royal College of Surgeons of England on 25th October, 1949. PMID- 19309886 TI - Hypertension: Hunterian Lecture delivered at the Royal College of Surgeons of England on 29th June, 1949. PMID- 19309887 TI - Splenectomy: Lecture delivered at the Royal College of Surgeons of England on 6th October, 1949. PMID- 19309888 TI - Excision of the Rectum for Carcinoma: Lecture delivered at the Royal College of Surgeons of England. PMID- 19309889 TI - Urine Hormones: Lecture delivered at the Royal College of Surgeons of England on 2nd May, 1949. PMID- 19309891 TI - Sir Arthur Sims. PMID- 19309890 TI - Infra-red Photographic Studies of the Superficial Veins in the Female and their Clinical Application: Arris and Gale Lecture delivered at the Royal College of Surgeons of England on 24th March, 1949. PMID- 19309892 TI - "Observables" at the Royal College of Surgeons: 27. The Grace Cup. PMID- 19309893 TI - The Journal of Bone and Joint Surgery. PMID- 19309894 TI - 150th Anniversary of the College. PMID- 19309895 TI - "Observables" at the Royal College of Surgeons: 28. Charters. The Charter Of Incorporation Of The Barbers' Company. PMID- 19309896 TI - Grant of Fellowship Diplomas. PMID- 19309897 TI - Sims Commonwealth Professor in Canada. PMID- 19309898 TI - "Observables" at the Royal College of Surgeons: 30. The Tipstaff Of The Company Of Surgeons. PMID- 19309900 TI - Lincoln Green: Reprinted, by kind permission, from the Times of September 4, 1950. PMID- 19309899 TI - Third Anniversary of the Faculty of Dental Surgeons. PMID- 19309901 TI - Presentation of Honorary Medal of The College to Lord Webb-Johnson. PMID- 19309902 TI - "Observables" at the Royal College of Surgeons: 32. Silver Tankard Presented by the Royal Australasian College of Surgeons. PMID- 19309903 TI - View Day. PMID- 19309905 TI - The Celebration of the 25th Anniversary of the Foundation of the Fouad First University, Cairo, and its Medical Faculty. PMID- 19309904 TI - Grant of Fellowship Diploma. PMID- 19309906 TI - Dinner to Sir William Kelsey Fry. PMID- 19309907 TI - Imperial Cancer Research Fund. PMID- 19309909 TI - Presentation of Medals to Blood Donors by H.R.H. the Princess Royal. PMID- 19309908 TI - Lister Medal. PMID- 19309910 TI - A Record of the Years from 1901 to 1950. PMID- 19309911 TI - Dr. Howard K. Gray of the Mayo Clinic. PMID- 19309912 TI - The New College Book-Plate. PMID- 19309913 TI - Death of Professor George Grey Turner, F.R.C.S. PMID- 19309914 TI - "Observables" at the Royal College of Surgeons: 37. Badges for Past Presidents. PMID- 19309915 TI - Faculty of Anaesthetists. PMID- 19309916 TI - Appointment of Fellows and Members to Consultant Posts. PMID- 19309917 TI - Appointment of Fellows and Members to Consultant Posts. PMID- 19309918 TI - The Thomas Vicary Commemoration. PMID- 19309919 TI - "Observables" at the Royal College of Surgeons: 38. The Master's Chair. PMID- 19309920 TI - Appointment of Fellows and Members to Consultant Posts. PMID- 19309921 TI - Acute Cardiac Arrest. PMID- 19309922 TI - Visit of H.R.H. The Princess Elizabeth to Receive the Honorary Fellowship. PMID- 19309924 TI - Memorial to John Hunter and Frank Buckland. PMID- 19309923 TI - College Council Club Ladies' Dinner. PMID- 19309925 TI - Increased Subscription Rate. PMID- 19309926 TI - His late Majesty King George VI. PMID- 19309927 TI - Viscount Addison: An Appreciation by Lord Webb-Johnson in the House of Lords on 30th January, 1952. PMID- 19309929 TI - The College as a Charity. PMID- 19309928 TI - The Hallett Prize. PMID- 19309931 TI - "Long Calderwood" (Home of the Hunters). PMID- 19309930 TI - The Listerian Festival. PMID- 19309933 TI - Restoration and Rebuilding of the College. PMID- 19309932 TI - Restoration and Rebuilding of the College. PMID- 19309934 TI - Restoration and Rebuilding of the College. PMID- 19309935 TI - The Lister Collection of Surgical Instruments. PMID- 19309936 TI - The Buckston Browne Benefaction. PMID- 19309937 TI - Restoration and Rebuilding of the College. PMID- 19309938 TI - The Jubilee of the Imperial Cancer Research Fund. PMID- 19309939 TI - College Tie. PMID- 19309940 TI - Restoration and Rebuilding of the College. PMID- 19309942 TI - Restoration and Rebuilding of the College. PMID- 19309943 TI - Two Pioneer Surgeons. PMID- 19309945 TI - The MacRae-Webb-Johnson Fund. PMID- 19309944 TI - Restoration and Rebuilding of the College. PMID- 19309947 TI - Restoration and Rebuilding of the College. PMID- 19309946 TI - The President Receiving the Diploma of Honorary Fellowship of the Royal College of Surgeons of Edinburgh. PMID- 19309948 TI - Restoration and Rebuilding of the College. PMID- 19309949 TI - Restoration and Rebuilding of the College. PMID- 19309950 TI - Primary Examination for the Fellowship. PMID- 19309951 TI - Hunterian Festival, 1953. PMID- 19309952 TI - Restoration and Rebuilding of the College. PMID- 19309953 TI - July Issue. PMID- 19309954 TI - The new College Bookplate. PMID- 19309956 TI - The New College Bookplate. PMID- 19309955 TI - Restoration and Rebuilding of the College. PMID- 19309958 TI - Restoration and Rebuilding of the College. PMID- 19309957 TI - Restoration and Rebuilding of the College. PMID- 19309960 TI - Visit of H.R.H. The Duke of Edinburgh to the College to Receive the Honorary Fellowship. PMID- 19309959 TI - Restoration and Rebuilding of the College. PMID- 19309962 TI - Lives of the Fellows 1930-51. PMID- 19309961 TI - Restoration and Rebuilding of the College. PMID- 19309963 TI - Restoration and Rebuilding of the College. PMID- 19309965 TI - Lives of the Fellows 1930-1951. PMID- 19309964 TI - Chairs in the Great Hall. PMID- 19309967 TI - Clinical Conferences. PMID- 19309966 TI - Restoration and Rebuilding of the College. PMID- 19309968 TI - Raffaele Bastianelli. PMID- 19309969 TI - Mount Everest Evening. PMID- 19309971 TI - Fellows of Fifty Years' Standing. PMID- 19309970 TI - Restoration and Rebuilding of the College. PMID- 19309972 TI - President's Visit to Ceylon and the Middle East. PMID- 19309973 TI - Restoration and Rebuilding of the College. PMID- 19309974 TI - The Trustees of the Hunterian Collection. PMID- 19309975 TI - The Mitchiner and Lady Cade Medals. PMID- 19309976 TI - Robert Jones National Memorial Fund Travelling Research Fellowship in Orthopaedic Surgery. PMID- 19309977 TI - Sir Frank Colyer, K.B.E., F.R.C.S., F.D.S. PMID- 19309978 TI - Restoration and Rebuilding of the College. PMID- 19309979 TI - Restoration and Rebuilding of the College. PMID- 19309980 TI - The Bicentenary Celebrations of the Royal Society of Arts. PMID- 19309982 TI - Gifts of Chairs. PMID- 19309981 TI - A Supplementary Note on Nelaton's Probe. PMID- 19309983 TI - Unveiling the Eagle. PMID- 19309984 TI - Bis Dat Qui Cito Dat. PMID- 19309985 TI - Parliamentary and Scientific Committee. PMID- 19309986 TI - University of Leeds-Congregation for the Conferment of Honorary Degrees. PMID- 19309987 TI - Restoration and Rebuilding of the College. PMID- 19309988 TI - Gifts for Furnishing. PMID- 19309989 TI - Restoration and Rebuilding of the College. PMID- 19309990 TI - Restoration and Rebuilding of the College. PMID- 19309991 TI - The Lister Medal. PMID- 19309993 TI - Primary Fellowship Examination Overseas. PMID- 19309992 TI - City Companies. PMID- 19309995 TI - Sir Harry Platt, M.D., M.S., P.R.C.S. PMID- 19309994 TI - The Late Sergei Sergevitch Yudin, Hon. F.R.C.S. PMID- 19309996 TI - Restoration and Rebuilding of the College. PMID- 19309997 TI - Back Numbers of the Annals. PMID- 19309998 TI - Professor Frederic Wood Jones, D.Sc., F.R.S., F.R.C.S. PMID- 19309999 TI - Restoration and Rebuilding of the College. PMID- 19310000 TI - Gifts of Chairs. PMID- 19310001 TI - Back Numbers of the Annals. PMID- 19310002 TI - Restoration and Rebuilding of the College. PMID- 19310004 TI - Hospitals' Symphony Orchestra. PMID- 19310003 TI - The New Library Extension. PMID- 19310005 TI - Barber-Surgeons Again. PMID- 19310006 TI - The Spider and the Poulterer: A Yarn of the Spun. PMID- 19310007 TI - Restoration and Rebuilding of the College. PMID- 19310008 TI - Restoration and Rebuilding of the College. PMID- 19310009 TI - Back Numbers of the Annals. PMID- 19310010 TI - Binding of the Annals. PMID- 19310011 TI - Restoration and Rebuilding of the College. PMID- 19310012 TI - Back Numbers of the Annals. PMID- 19310013 TI - American College of Surgeons Visits Jamaica. PMID- 19310014 TI - Sir Arthur Keith. PMID- 19310016 TI - Back Numbers of the Annals. PMID- 19310015 TI - Restoration and Rebuilding of the College. PMID- 19310017 TI - Restoration and Rebuilding of the College. PMID- 19310019 TI - Restoration and Rebuilding of the College. PMID- 19310018 TI - Back Numbers of the Annals. PMID- 19310020 TI - Binding of the Annals. PMID- 19310021 TI - Restoration and Rebuilding of the College. PMID- 19310023 TI - Sir Arthur Sims Commonwealth Travelling Professorship. PMID- 19310022 TI - Gifts of Chairs and Tables. PMID- 19310025 TI - Presentations. PMID- 19310024 TI - Lectures by John Hunter. PMID- 19310026 TI - Restoration and Rebuilding of the College. PMID- 19310027 TI - William Clift. PMID- 19310028 TI - Heroes of the Examination Book: 1. James Noble (1774-1851). PMID- 19310029 TI - Society of Apothecaries of London. PMID- 19310030 TI - Lives of the Fellows 1930-51. PMID- 19310031 TI - Restoration and Rebuilding of the College. PMID- 19310032 TI - Heroes of the Examination Book: 2. James Macnamara (1768-1826). PMID- 19310033 TI - Restoration and Rebuilding of the College. PMID- 19310034 TI - George Bennett, F.R.C.S. (1804-1893). PMID- 19310035 TI - The Anatomist's Medal of the Company of Surgeons. PMID- 19310036 TI - Heroes of the Examination Book: 3. Sir Charles Napier (1786-1860). PMID- 19310037 TI - The President visits the United States and Canada. PMID- 19310038 TI - Visit of Soviet Doctors. PMID- 19310040 TI - Primary Fellowship in Colombo. PMID- 19310039 TI - Restoration and Rebuilding of the College. PMID- 19310041 TI - Restoration and Rebuilding of the College. PMID- 19310042 TI - Thomas Vicary Commemoration. PMID- 19310043 TI - Visit to the United States and Canada. PMID- 19310045 TI - Gifts of Chairs and Tables. PMID- 19310044 TI - William Clift. PMID- 19310047 TI - Primary Fellowship in Ceylon 1956. PMID- 19310046 TI - Heroes of the Examination Book: 4. The Honourable George Cranfield Berkeley (1753 1818). PMID- 19310049 TI - Restoration and Rebuilding of the College. PMID- 19310048 TI - Increase in Subscription. PMID- 19310050 TI - Sims Commonwealth Travelling Professors for 1957. PMID- 19310051 TI - Increase in Subscription. PMID- 19310052 TI - A Visit of Professor Carus of Dresden to the Royal College of Surgeons in 1844. PMID- 19310053 TI - Visit of Soviet Surgeons to the College. PMID- 19310054 TI - Dr. W. D. M. Paton, F.R.S. PMID- 19310056 TI - The College Tie. PMID- 19310055 TI - Restoration and Rebuilding of the College. PMID- 19310057 TI - Restoration and Rebuilding of the College. PMID- 19310058 TI - Restoration and Rebuilding of the College. PMID- 19310059 TI - Visit of Representatives of the American College of Surgeons to the College 18th 21st April 1956. PMID- 19310060 TI - The Mutiny at the Nore. PMID- 19310061 TI - The Cassell Building. PMID- 19310062 TI - Imperial Cancer Research Fund. PMID- 19310063 TI - Binding of the Annals. PMID- 19310064 TI - R.L.S. and G.B.B. PMID- 19310065 TI - Reunion with the Academie de Chirurgie: May 31st-June 1st 1956. PMID- 19310066 TI - Restoration and Rebuilding of the College: The Nuffield College. PMID- 19310067 TI - Tour of the East. PMID- 19310068 TI - Overseas Visitors to the College. PMID- 19310069 TI - Overseas Visitors to the College. PMID- 19310071 TI - Restoration and Rebuilding of the College. PMID- 19310070 TI - Sir Gordon Gordon-Taylor, K.B.E., C.B., F.R.C.S. PMID- 19310072 TI - Postgraduate Education Brochure. PMID- 19310073 TI - Sims Commonwealth Travelling Professor. PMID- 19310075 TI - Overseas Visitors to the College. PMID- 19310074 TI - Delegation of Surgeons to the U.S.S.R. PMID- 19310076 TI - Surgical Technique-Homage to Arbuthnot Lane. PMID- 19310077 TI - Nuffield Research Chair in Dental Science. PMID- 19310078 TI - Restoration and Rebuilding of the College. PMID- 19310079 TI - Lister Medal. PMID- 19310080 TI - Long Calderwood. PMID- 19310082 TI - Restoration and Rebuilding of the College. PMID- 19310081 TI - Gift from Sir Gordon Gordon-Taylor. PMID- 19310083 TI - Fiftieth Anniversary of the Admission to the Fellowship of Lord Webb-Johnson and Sir Gordon Gordon-Taylor. PMID- 19310084 TI - Banker's Orders. PMID- 19310086 TI - Sir Gordon Gordon-Taylor, K.B.E., C.B., F.R.C.S. PMID- 19310085 TI - Restoration and Rebuilding of the College. PMID- 19310088 TI - Overseas Visitors to the College. PMID- 19310087 TI - Restoration and Rebuilding of the College. PMID- 19310089 TI - The Hunterian Festival. PMID- 19310091 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310090 TI - Five Rings and a Roundabout. PMID- 19310092 TI - Overseas Visitors to the College. PMID- 19310093 TI - Penrose-May Tutorial Fund. PMID- 19310094 TI - Visit of the Academie de Chirurgie de Paris. PMID- 19310095 TI - Hunter House Memorial Fund. PMID- 19310096 TI - Overseas Visitors to the College. PMID- 19310097 TI - Anatomical Material. PMID- 19310098 TI - Restoration and Rebuilding of the College. PMID- 19310099 TI - College Grace. PMID- 19310101 TI - Sir James Paterson Ross-President. PMID- 19310100 TI - Seats of Learning. PMID- 19310102 TI - Restoration and Rebuilding of the College. PMID- 19310103 TI - Annals Editorial Office. PMID- 19310104 TI - Supplemental Charter. PMID- 19310105 TI - Association of Surgeons of India. PMID- 19310107 TI - Opening of the Department of Anaesthetics. PMID- 19310106 TI - Liver Function Tests in Relation to Surgery. PMID- 19310108 TI - Restoration and Rebuilding of the College: Audio-Visual Systems-Great Hall. PMID- 19310109 TI - The 21st Volume of the Annals. PMID- 19310110 TI - Sir James Paget's Notes on His Students. PMID- 19310111 TI - Annals Editorial Office. PMID- 19310112 TI - Anterior Surgical Approaches to the Spine. PMID- 19310113 TI - Annals Editorial Office. PMID- 19310115 TI - The Coulthurst Room. PMID- 19310114 TI - Restoration and Rebuilding of the College. PMID- 19310117 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310116 TI - Overseas Visitors to the College. PMID- 19310119 TI - The Thomas Vicary Commemoration. PMID- 19310118 TI - Overseas Visitors to the College. PMID- 19310120 TI - Sir Simon Marks, Hon. F.R.C.S: A Great Benefactor. PMID- 19310121 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310123 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310122 TI - Restoration and Rebuilding of the College. PMID- 19310124 TI - Primary Fellowship Examination in Ceylon. PMID- 19310125 TI - Sir Harry Platt, Bt. PMID- 19310126 TI - The 1957 Charter. PMID- 19310127 TI - Restoration and Rebuilding of the College. PMID- 19310128 TI - Overseas Visitors to the College. PMID- 19310130 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310129 TI - Sir James Walton. PMID- 19310131 TI - Recent Overseas Visitors to the College. PMID- 19310132 TI - Course in Statistics and Radio-Isotopes: 5th to 9th May 1958. PMID- 19310134 TI - The New Anatomical Nomenclature. PMID- 19310133 TI - Curiosities of Natural History. PMID- 19310136 TI - The Lister Festival. PMID- 19310135 TI - Diploma Granting Ceremony. PMID- 19310137 TI - Admission to the Honorary Fellowship of Professor C. Crafoord. PMID- 19310139 TI - LIFE OF SIR GEORGE BUCKSTON BROWNE, F.R.C.S. PMID- 19310138 TI - Letters of Abernethy. PMID- 19310140 TI - Some Happy Memories. PMID- 19310141 TI - The Rt. Hon. Lord Webb-Johnson. PMID- 19310143 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310142 TI - From Trade Guild to Royal College: The Inaugural Address delivered to the Abernethian Society of St. Bartholomew's Hospital on 3rd October 1957. PMID- 19310144 TI - Overseas Visitors to the College. PMID- 19310145 TI - Binding of the Annals. PMID- 19310146 TI - Restoration and Rebuilding of the College. PMID- 19310148 TI - Imperial Cancer Research Fund. PMID- 19310147 TI - Recent Overseas Visitors to the College. PMID- 19310149 TI - On Examining John Hunter's Pathological Specimens. PMID- 19310150 TI - Recent Overseas Visitors. PMID- 19310151 TI - The Buckston Browne Benefaction. PMID- 19310152 TI - Recent Overseas Visitors to the College. PMID- 19310153 TI - Imperial Cancer Research Fund. PMID- 19310154 TI - College Council Club. PMID- 19310156 TI - Hand Surgery. PMID- 19310155 TI - Restoration and Rebuilding. PMID- 19310157 TI - Imperial Cancer Research Fund. PMID- 19310158 TI - Guildford Cathedral. PMID- 19310159 TI - Hand Surgery. PMID- 19310160 TI - Sir Harry Platt, Bt. PMID- 19310161 TI - Recent Overseas Visitors to the College. PMID- 19310162 TI - Surgeons sine sede. PMID- 19310163 TI - Recent Overseas Visitors to the College. PMID- 19310164 TI - The Thomas Vicary Commemoration. PMID- 19310165 TI - A Recent Addition to the College Treasures. PMID- 19310166 TI - The Royal College of Surgeons and Imperial Cancer Research Fund: Preliminary Contract. Underpinning and Demolition. PMID- 19310167 TI - Trustees of the Hunterian Collection. PMID- 19310168 TI - Medical Faculty of the University of the Punjab. PMID- 19310169 TI - Centenary of the First Publication of Gray's Anatomy. PMID- 19310171 TI - Recent Overseas Visitors to the College. PMID- 19310170 TI - The College Tie. PMID- 19310172 TI - The College Tie. PMID- 19310173 TI - Imperial Cancer Research Fund. PMID- 19310174 TI - Letters of John Hunter. PMID- 19310176 TI - Recent Overseas Visitors to the College. PMID- 19310175 TI - Centenary Celebrations of the Royal Charter Instituting The Licence in Dental Surgery: 22nd, 23rd, 24th July 1959. PMID- 19310178 TI - A Consultation with Boerhaave. PMID- 19310177 TI - John Hunter's Giraffe. PMID- 19310179 TI - Recent Overseas Visitors to the College. PMID- 19310180 TI - Sir Harry Platt, Bt. PMID- 19310182 TI - The College Tie. PMID- 19310181 TI - John Hunter's Statue. PMID- 19310183 TI - The Hunterian Festival. PMID- 19310185 TI - Fellows' Annual Subscriptions United Kingdom Income Tax Relief. PMID- 19310184 TI - Sir Harry Platt, Bt. PMID- 19310186 TI - Life in Emergency Ward 10. PMID- 19310187 TI - Life of Sir George Buckston Browne, F.R.C.S. PMID- 19310189 TI - Binding of the Annals. PMID- 19310188 TI - Cardiac Surgery. PMID- 19310190 TI - Voluntary Annual Subscriptions. PMID- 19310191 TI - Recent Overseas Visitors to the College. PMID- 19310193 TI - Cardiac Surgery. PMID- 19310192 TI - Life in Emergency Ward 10. PMID- 19310194 TI - Binding of the Annals. PMID- 19310195 TI - Recent Overseas Visitors to the College. PMID- 19310196 TI - Centenary Celebrations of the Royal Charter instituting the Licence in Dental Surgery: 22nd, 23rd and 24th July, 1959. PMID- 19310197 TI - Howard Gray Memorial Library. PMID- 19310198 TI - The Printing Strike. PMID- 19310200 TI - The Fellows' Common Room. PMID- 19310199 TI - Recent Overseas Visitors to the College. PMID- 19310201 TI - Guildford Cathedral. PMID- 19310202 TI - Recent Visitors. PMID- 19310203 TI - Hand Surgery. PMID- 19310204 TI - Birthday Honours. PMID- 19310206 TI - A History of the Royal College of Surgeons of England. PMID- 19310205 TI - The College of General Practitioners and Lincoln's Inn Fields. PMID- 19310207 TI - Howard Gray Memorial Library. PMID- 19310208 TI - Guildford Cathedral: Doctors' Window. PMID- 19310209 TI - Past Presidents: Sir William Lawrence, Bart. PMID- 19310210 TI - Treatment of the Cleft Palate: Scientific Symposium: (b) The Surgery of Cleft Lip and Palate. PMID- 19310211 TI - Treatment of the Cleft Palate: Scientific Symposium: (d) Prosthetic Treatment. PMID- 19310213 TI - L.D.S. Centenary Celebrations: 22nd-24th July. PMID- 19310212 TI - Formal Opening of the L.D.S. Centenary Celebrations: At the Royal College of Surgeons of England on 23rd July 1959. PMID- 19310214 TI - L.D.S. Centenary Celebrations: Congratulatory Addresses. PMID- 19310215 TI - Howard Gray Memorial Library. PMID- 19310216 TI - Gifts to the Library. PMID- 19310217 TI - Conferment of Honorary Fellowship of the Royal College of Surgeons of Edinburgh. PMID- 19310218 TI - Imperial Cancer Research Fund. PMID- 19310219 TI - Sir Arthur Sims Commonwealth Travelling Professorship. PMID- 19310220 TI - Binding of the Annals. PMID- 19310221 TI - The College Tie. PMID- 19310222 TI - Hunterian Trustees. PMID- 19310223 TI - A New Catalogue of Portraits. PMID- 19310225 TI - A New Catalogue of Portraits. PMID- 19310224 TI - Carcinoid Tumours (Argentaffinomata): Hunterian Lecture delivered at the Royal College of Surgeons of England on 3rd March 1959. PMID- 19310227 TI - The Printing of the Annals. PMID- 19310226 TI - Thomas Vicary Commemoration. PMID- 19310229 TI - A New Catalogue of Portraits. PMID- 19310228 TI - Admission of Lord Kindersley to the Honorary Fellowship. PMID- 19310231 TI - Imperial Cancer Research Fund. PMID- 19310230 TI - Back Numbers of the Annals. PMID- 19310232 TI - Recent Overseas Visitors to the College. PMID- 19310233 TI - The Hunterian Trustees. PMID- 19310234 TI - Primary Fellowship Examinations Overseas. PMID- 19310235 TI - Hydatid Disease Affecting the Nervous System: Hunterian Lecture delivered at the Royal College of Surgeons of England on 7th October 1959. PMID- 19310236 TI - Recent Overseas Visitors to the College. PMID- 19310237 TI - Ruscoe Clarke Memorial Lecture. PMID- 19310238 TI - The College Appeal. PMID- 19310239 TI - The Cellular Anatomy of Experimental Wound Healing: Arris and Gale Lecture delivered at the Royal College of Surgeons of England on 17th December 1959. PMID- 19310241 TI - Binding of the Annals. PMID- 19310240 TI - Recent Overseas Visitors to the College. PMID- 19310242 TI - Sir Harry Platt, Bt. PMID- 19310243 TI - Some Edinburgh Pioneers in Orthopaedic Surgery: Robert Jones Lecture delivered at the Royal College of Surgeons of England on 14th January 1960. PMID- 19310244 TI - Sir Arthur Sims Commonwealth Travelling Professorships. PMID- 19310245 TI - Bernard Johnson Memorial Fund. PMID- 19310246 TI - Court of Patrons. PMID- 19310247 TI - Hospital Medical Staffs Defence Trust. PMID- 19310248 TI - Recent Overseas Visitors to the College. PMID- 19310250 TI - The Ether Painting. PMID- 19310249 TI - Portrait of the late Sir Archibald McIndoe. PMID- 19310251 TI - Admission to the Fellowship by Election. PMID- 19310252 TI - Hand Surgery. PMID- 19310253 TI - Sir William Coates, K.C.B., C.B.E., V.D., T.D., F.R.C.S. PMID- 19310255 TI - Overseas Examinations. PMID- 19310254 TI - The late Sir Archibald McIndoe, C.B.E., F.R.C.S. PMID- 19310256 TI - The Wilson-Barkworth Fund. PMID- 19310257 TI - Rebuilding of the College: Progress Report. PMID- 19310258 TI - The Honorary Medal of the College. PMID- 19310259 TI - Webb-Johnson Lecture. PMID- 19310260 TI - The Late Professor Seymour Barling. PMID- 19310261 TI - Paget's Surgical Pathology. PMID- 19310262 TI - Sir James Paterson Ross, Bt., K.C.V.O. PMID- 19310263 TI - Some Memories of a Fellow of 1896. PMID- 19310264 TI - The Darwin Medal. PMID- 19310266 TI - Sir Harry Platt, Bt. PMID- 19310265 TI - A Pilgrimage to Cos. PMID- 19310268 TI - Holiday Lectures for Schoolchildren. PMID- 19310267 TI - Surgical Treatment of Mandibular Prognathism: Hunterian Lecture delivered at the Royal College of Surgeons of England on 4th April 1960. PMID- 19310269 TI - The late Sir Gordon Gordon-Taylor. PMID- 19310271 TI - The College Tie. PMID- 19310270 TI - A Letter from John Hunter. PMID- 19310272 TI - Binding of the Annals. PMID- 19310274 TI - Lister Medal. PMID- 19310273 TI - Genetics in Relation to Surgery: A Historical Review: Thomas Vicary Lecture delivered at the Royal College of Surgeons of England on 27th October 1960. PMID- 19310275 TI - Bernard Johnson Memorial. PMID- 19310277 TI - Professor Raffaele Bastianelli. PMID- 19310276 TI - Natural History of Arteriosclerosis of the Lower Extremities: Hunterian Lecture delivered at the Royal College of Surgeons of England on 22nd April 1960. PMID- 19310278 TI - The Late Sir Gordon Gordon-Taylor: An appreciation from the Royal Australasian College of Surgeons. PMID- 19310279 TI - Imperial Cancer Research Fund. PMID- 19310280 TI - Response to the Broadcast by Sir Cecil Wakeley, Bt. PMID- 19310281 TI - Mr. Tipple's Chest Wound: A Case of Lateral Transfixion in the Year 1812. PMID- 19310282 TI - Rebuilding of the College: Progress Report: Phase IIIb. PMID- 19310283 TI - Retirement of Sir Wilfred Fish, C.B.E. PMID- 19310284 TI - A Visit to Pakistan. PMID- 19310285 TI - Imperial Cancer Research Fund: The New Laboratories. PMID- 19310286 TI - The Hunterian Festival. PMID- 19310287 TI - Fellows' Annual Subscriptions: United Kingdom Income Tax Relief. PMID- 19310288 TI - Surgical Theses. PMID- 19310289 TI - Hunterian Society of London. PMID- 19310290 TI - The Wolfson Foundation. PMID- 19310291 TI - The Cartwright Prize. PMID- 19310292 TI - Passages from the Notebooks of Sir Frederick Treves, Bt. PMID- 19310293 TI - The Training of Surgeons. PMID- 19310294 TI - The College Appeal. PMID- 19310295 TI - Osler Club. PMID- 19310296 TI - Museum, Research and the Inspiration of Hunter: Hunterian Oration delivered at the Royal College of Surgeons of England on 14th February 1961. PMID- 19310297 TI - Carcinoma of the Oesophagus: Reflections upon Surgical Treatment: Cecil Joll Lecture delivered at the Royal College of Surgeons of England on 28th July 1960. PMID- 19310298 TI - The Lister Festival. PMID- 19310299 TI - Activation of the Record of Human Experience: Summary of the Lister Oration delivered at the Royal College of Surgeons of England on 27th April 1961. PMID- 19310300 TI - Malignant Melanoma of the Choroid treated with Radioactive Applicators: Hunterian Lecture delivered at the Royal College of Surgeons of England on 29th November 1960. PMID- 19310302 TI - Sims Commonwealth Travelling Professor. PMID- 19310301 TI - Gift of Miniatures. PMID- 19310303 TI - Guildford Cathedral. PMID- 19310304 TI - Gordon-Taylor Memorial Lecture. PMID- 19310305 TI - Passages from the Notebook of Sir Frederick Treves, Bt: II. The Case of Sir Henry Irving. PMID- 19310306 TI - Sims Commonwealth Travelling Professor. PMID- 19310307 TI - The Bland Sutton Lecturer. PMID- 19310308 TI - The College Tie. PMID- 19310309 TI - Binding of the Annals. PMID- 19310311 TI - Primary Fellowship Examinations Overseas. PMID- 19310310 TI - A Visit to Khartoum: A resume of the Report. PMID- 19310312 TI - Increase in Subscription Rate. PMID- 19310313 TI - A Small Library: The Syme Oration delivered at the opening ceremony of the meeting of the Royal Australasian College of Surgeons at Brisbane on 23rd May 1961. PMID- 19310314 TI - Christmas Holiday Lectures for Schoolchildren. PMID- 19310315 TI - Centenary Memorials. PMID- 19310316 TI - The Epitome of Vesalius: Another notable acquisition for the Library. PMID- 19310317 TI - McNeill Love Medal: Recognition of Long Service in the College. PMID- 19310318 TI - Microfilm Edition of the Annals. PMID- 19310319 TI - Bernard Darwin and Downe. PMID- 19310320 TI - Thomas Vicary Commemoration. PMID- 19310321 TI - The Conservators of the Hunterian Museum: I. William Clift. PMID- 19310323 TI - The Conservators of the Hunterian Museum: II. Richard Owen. PMID- 19310322 TI - A Collection of Wedgwood Letters. PMID- 19310325 TI - In Outer Space. PMID- 19310324 TI - Binding of the Annals. PMID- 19310326 TI - Gordon-Taylor Memorial Lecture. PMID- 19310327 TI - Conservators of the Hunterian Museum: III. John Thomas Quekett. PMID- 19310328 TI - Course on the Uses of Data Handling and Computing. PMID- 19310329 TI - The Overseas Primary F.R.C.S. PMID- 19310330 TI - Twenty-Five Years' Service at the College: Sidney Charles Bartlett: Charles George Bush Edward John Noon. PMID- 19310331 TI - Institute of Basic Medical Sciences: Royal College of Surgeons of England British Postgraduate Medical Federation (University of London). PMID- 19310332 TI - Moynihan: The Education and Training of the Surgeon: Eleventh Moynihan Lecture delivered at the University of Leeds on 25th May 1961. PMID- 19310333 TI - In My End Is My Beginning: Frederic Hewitt Lecture delivered at the Royal College of Surgeons of England on 15th March 1961. PMID- 19310334 TI - Twenty-Five Years' Service at the College: Mr. W. C. Willmott Mrs. K. Green. PMID- 19310335 TI - The Overseas Primary F.R.C.S. PMID- 19310336 TI - Imperial Cancer Research Fund: Cancer Week 1962. PMID- 19310337 TI - Conservators of the Hunterian Museum: IV. William Henry Flower. PMID- 19310338 TI - Fellows' Annual Subscriptions: United Kingdom Income Tax Relief. PMID- 19310339 TI - Conservators of the Hunterian Museum: V. Charles Stewart. PMID- 19310340 TI - Professor Digby Chamberlain, C.B.E. PMID- 19310341 TI - Centenary of the Death of Thomas Wakley, M.R.C.S. PMID- 19310343 TI - Sir Harry Platt, Bt. PMID- 19310342 TI - Twenty-five Years' Service at the College. PMID- 19310344 TI - Reminiscences of Sir Victor Horsley. PMID- 19310345 TI - 31st Volume of the Annals. PMID- 19310346 TI - Surgical Theses. PMID- 19310347 TI - Visit of the President to Africa. PMID- 19310348 TI - Completion of Building Operations. PMID- 19310349 TI - To Acknowledge a Generous and Welcome Gift from Sir Victor Negus. PMID- 19310350 TI - The Surgical Rehabilitation of the Rheumatoid Hand: Hunterian Lecture delivered at the Royal College of Surgeons of England on 14th June 1962. PMID- 19310351 TI - An African Surgical Pilgrimage. PMID- 19310352 TI - Jean-Baptiste Huet. PMID- 19310353 TI - Research in Orthopaedics. PMID- 19310354 TI - Imperial Cancer Research Fund. PMID- 19310356 TI - The College Tie. PMID- 19310355 TI - On the Pathogenesis of Mixed Anaerobic Infections of Mucous Membranes: Charles Tomes Lecture delivered at the Royal College of Surgeons of England on 20th July 1962. PMID- 19310357 TI - Retirement of Mr. Kennedy Cassels. PMID- 19310358 TI - Memorial Service for Professor Ian Aird: Thursday, 11th October 1962, St. Columba's, Pont Street. PMID- 19310360 TI - Porritt Working Party on Medical Aid to the Developing Countries. PMID- 19310359 TI - Binding of the Annals. PMID- 19310361 TI - Overseas Examinations. PMID- 19310363 TI - Sir James Berry's Gold Medal. PMID- 19310362 TI - Complications of Cholecystectomy: Lecture delivered at the Royal College of Surgeons of England on 25th September 1962. PMID- 19310365 TI - Research Defence Society. PMID- 19310364 TI - University of the Panjab. PMID- 19310366 TI - The Seventh British Empire and Commonwealth Games. PMID- 19310368 TI - International Federation of Surgical Colleges. PMID- 19310367 TI - "Si Monumentum Requiris Circumspice": Plastic Surgery in War and Peace: The first McIndoe Lecture delivered at the Royal College of Surgeons of England on 7th December 1962. PMID- 19310369 TI - The Basal Ganglia and Locomotion: Arris and Gale Lecture delivered at the Royal College of Surgeons of England on 3rd January 1963. PMID- 19310371 TI - The Hunterian Festival. PMID- 19310370 TI - Improvements and Additions to "Anatomical Techniques": IV. Casts of the Cerebrospinal Fluid System of the Brain. PMID- 19310373 TI - Overseas Primary Examinations. PMID- 19310372 TI - International Federation of Surgical Colleges. PMID- 19310374 TI - Committee of Management. PMID- 19310375 TI - Heroes of the Examination Books: 5. Sir Thomas Francis Fremantle (1765-1819). PMID- 19310376 TI - Re-opening of the Hunterian Museum. PMID- 19310377 TI - Laming Evans Senior Research Fellowship: The Royal College of Surgeons of England in association with Makerere University College. PMID- 19310378 TI - Visit of H.R.H. The Princess Margaret to receive Honorary Fellowship. PMID- 19310379 TI - Overseas Primary Examinations. PMID- 19310380 TI - Completion of the Hunterian Museum. PMID- 19310382 TI - Memorial Plaque. PMID- 19310381 TI - Convention on Accident Prevention and Life Saving. PMID- 19310383 TI - Faculty of Anaesthetists. PMID- 19310384 TI - Diploma Granting Ceremony. PMID- 19310385 TI - Visit of H.M. The Queen to The Royal Marsden Hospital. PMID- 19310386 TI - Members of the Council. PMID- 19310387 TI - International Federation of Surgical Colleges. PMID- 19310388 TI - Imperial Cancer Research Fund. PMID- 19310389 TI - Hunterian Museum. PMID- 19310390 TI - Lister Medal. PMID- 19310391 TI - General Pathology. PMID- 19310392 TI - Rohan Williams Memorial Fund. PMID- 19310393 TI - Thomas Vicary Commemoration. PMID- 19310394 TI - Heroes of the Examination Books: 6. Sir Thomas Boulden Thompson. PMID- 19310395 TI - The Trustees of the Hunterian Collection. PMID- 19310396 TI - Telstar Broadcast. PMID- 19310397 TI - The Court of Patrons. PMID- 19310399 TI - Heroes of the Examination Books: 7. John Harvey. PMID- 19310398 TI - Laming Evans Research Fund. PMID- 19310400 TI - Institute of Basic Medical Sciences. PMID- 19310401 TI - The Canadian Mace. PMID- 19310402 TI - Pilot Scheme for Surgical Tutors. PMID- 19310403 TI - Heroes of the Examination Books: 8. Sir Thomas Pasley. PMID- 19310404 TI - Introduction. PMID- 19310405 TI - The Site of the Operating Suite. PMID- 19310406 TI - Ratio of Operating Theatres to Surgical Beds. PMID- 19310408 TI - The Influence of Central Sterile Supply upon Theatre Design. PMID- 19310407 TI - The Size of the Theatres. PMID- 19310409 TI - Avoidance of Infection. PMID- 19310410 TI - Reception Area, Post-operative Recovery Unit and Intensive Therapy Ward. PMID- 19310411 TI - Anaesthetic Rooms. PMID- 19310412 TI - Viewing Domes, Galleries and Television. PMID- 19310413 TI - The Lighting of the Operating Theatre Suite. PMID- 19310414 TI - Rooms for Ancillary Services. PMID- 19310415 TI - Materials for Fixtures and Finishings. PMID- 19310416 TI - Sterile Water Supply. PMID- 19310417 TI - Sterilization by Steam under Pressure. PMID- 19310419 TI - Observations on Theatre Conditions in Warm Weather. PMID- 19310418 TI - Other Methods of Sterilization. PMID- 19310420 TI - Royal Photographic Society: Combined Royal Colleges Bronze Medal. PMID- 19310421 TI - The College Tie. PMID- 19310422 TI - The Lister Festival. PMID- 19310423 TI - Back Numbers of the Annals. PMID- 19310424 TI - Ceremony of Presentation of Diplomates. PMID- 19310425 TI - Council and Court Dinner. PMID- 19310426 TI - Binding of the Annals. PMID- 19310427 TI - The British Club for Surgery of the Hand. PMID- 19310428 TI - Sir Hugh Lett. Bt. PMID- 19310429 TI - Patterns of Intestinal Ischaemia: Arris and Gale Lecture delivered at the Royal College of Surgeons of England on 6th February 1964. PMID- 19310430 TI - Faculty of Anaesthetists. PMID- 19310431 TI - The Primary Fellowship Overseas. PMID- 19310432 TI - Heroes of the Examination Books: Sir Samuel Hood, Bt. PMID- 19310434 TI - Long Service Award. PMID- 19310433 TI - Thomas Vicary Commemoration. PMID- 19310435 TI - The College and the Royal Navy. PMID- 19310436 TI - Diploma Granting Ceremony. PMID- 19310438 TI - The College and the Royal Air Force. PMID- 19310437 TI - Royal College of Physicians. PMID- 19310439 TI - Binding of the Annals. PMID- 19310440 TI - British Journal of Surgery. PMID- 19310441 TI - "Observables" at the Royal College of Surgeons: 47. The Porritt Cup. PMID- 19310442 TI - Register of Fellows. PMID- 19310444 TI - Long Course of Tutorials, Lectures and Clinical Conferences. PMID- 19310443 TI - Final Fellowship Regulations. PMID- 19310445 TI - Short Course of Lectures and Clinical Conferences. PMID- 19310446 TI - Hunterian Festival. PMID- 19310447 TI - Admission to the Honorary Fellowship and to the Court of Patrons. PMID- 19310448 TI - Admission to the Court of Patrons. PMID- 19310449 TI - The College Tie. PMID- 19310450 TI - "The Surgeon's Craft". PMID- 19310451 TI - "Observables" at the Royal College of Surgeons: 48. Hunter's Prize Medal. PMID- 19310452 TI - The College Tie. PMID- 19310453 TI - Diploma Day. PMID- 19310455 TI - Lister's Writings. PMID- 19310454 TI - The British Club for Surgery of the Hand. PMID- 19310456 TI - Lister's Writings. PMID- 19310457 TI - Holiday Lectures for Young People. PMID- 19310458 TI - Lister Medal. PMID- 19310459 TI - Laming Evans Senior Research Fellowship. PMID- 19310460 TI - Lord brock of wimbledon. PMID- 19310461 TI - Sir Thomas Browne's Skull. PMID- 19310463 TI - The midland institute of otology. PMID- 19310462 TI - Lord brock of wimbledon. PMID- 19310464 TI - International Federation of surgical colleges. PMID- 19310465 TI - Department of dental science. PMID- 19310466 TI - Great Hall chairs. PMID- 19310468 TI - "Observables" at the Royal College of Surgeons: 49. Academical costume. PMID- 19310467 TI - New book. PMID- 19310469 TI - National army museum building appeal. PMID- 19310471 TI - The british club for surgery of the hand. PMID- 19310470 TI - The girdlestone tradition: inauguration of orthopaedic and traumatic services in the Sudan. PMID- 19310472 TI - British journal of surgery. PMID- 19310473 TI - Moynihan centenary volume. PMID- 19310474 TI - Diploma day. PMID- 19310475 TI - International corneo-plastic conference. PMID- 19310476 TI - Primary fellowship examination. PMID- 19310477 TI - Sir robert bradlaw, C.B.e. PMID- 19310479 TI - Sir arthur keith medal. PMID- 19310478 TI - Accommodation for outside functions. PMID- 19310480 TI - National army museum building appeal. PMID- 19310481 TI - Westminster abbey 900th anniversary year: special service for science, medicine and technology. PMID- 19310482 TI - I. Parathyroid Disease: 4. Parathyroid disease. PMID- 19310483 TI - College cricket. PMID- 19310484 TI - The worshipful society of apothecaries in london. PMID- 19310485 TI - Visit of the king of Thailand. PMID- 19310486 TI - Nobel prize in medicine. PMID- 19310488 TI - Letters from edward jenner. PMID- 19310487 TI - The worshipful company of barbers. PMID- 19310490 TI - Sir arthur porritt, bt. PMID- 19310489 TI - Holiday lectures for young people. PMID- 19310491 TI - The Thomas vicary commemoration. PMID- 19310492 TI - Sir harry platt, bt. PMID- 19310493 TI - John Hunter's Premonition of Death. PMID- 19310494 TI - Lister commemorative tablet. PMID- 19310496 TI - Post-war presidents. PMID- 19310495 TI - Surgery lectures. PMID- 19310497 TI - Council club dinner. PMID- 19310498 TI - Hunterian festival. PMID- 19310499 TI - Admission to the court of patrons. PMID- 19310500 TI - Final fellowship examination. PMID- 19310501 TI - Holbein cartoon. PMID- 19310503 TI - Hunterian trustees. PMID- 19310502 TI - Back issues of the annals. PMID- 19310504 TI - Freemasons fellowship fund. PMID- 19310505 TI - The british club for surgery of the hand. PMID- 19310506 TI - Holbein cartoon. PMID- 19310507 TI - Supplemental charter 1967. PMID- 19310508 TI - The primary examination. PMID- 19310509 TI - Back issues of the annals. PMID- 19310510 TI - The british club for surgery of the hand. PMID- 19310511 TI - Holbein cartoon. PMID- 19310512 TI - The worshipful society of apothecaries of london. PMID- 19310513 TI - Presentation of portrait of samuel cartwright, junior. PMID- 19310514 TI - Penrose may surgical tutor. PMID- 19310515 TI - The college tie. PMID- 19310516 TI - Holiday lectures for young people. PMID- 19310518 TI - International congress of cardiovascular surgery. PMID- 19310517 TI - Eightieth birthday. PMID- 19310520 TI - What the Undergraduate should know of Cancer. PMID- 19310519 TI - The role of plastic surgery in the treatment of malignant disease. PMID- 19310521 TI - The dental profession and cancer of the mouth and jaws. PMID- 19310522 TI - Endocrine aspects of advanced mammary cancer. PMID- 19310523 TI - Chemotherapy. PMID- 19310524 TI - The place of radiotherapy in the treatment of malignant disease. PMID- 19310525 TI - The prevention of cancer. PMID- 19310526 TI - Spontaneous regression of cancer. PMID- 19310527 TI - The care of the terminal stages of cancer. PMID- 19310528 TI - The legal aspects of cancer attributability. PMID- 19310529 TI - Summary. PMID- 19310531 TI - Control of pain: surgeon. PMID- 19310530 TI - Cancer in children. PMID- 19310532 TI - Control of pain: physician. PMID- 19310533 TI - Surgery in the inoperable cancer patient. PMID- 19310535 TI - Increase in subscription rate. PMID- 19310534 TI - The immunological approach to the treatment of cancer. PMID- 19310537 TI - The college tie. PMID- 19310536 TI - John Hunter's Last Pupil. PMID- 19310538 TI - Royal society of medicine. PMID- 19310539 TI - Research fellowship in biological engineering. PMID- 19310540 TI - The jacksonian prize. PMID- 19310541 TI - Editorship of the annals. PMID- 19310542 TI - Sir arthur porritt, bt. PMID- 19310543 TI - Jacksonian prize essay. PMID- 19310545 TI - A gift from malaysia. PMID- 19310544 TI - Retirement of mr. J. C. Higgins. PMID- 19310546 TI - Retirement of the librarian. PMID- 19310547 TI - The britain-Nepal medical expedition. PMID- 19310548 TI - Faraday exhibition. PMID- 19310549 TI - "Observables" at the Royal College of Surgeons: 52. Lord Moynihan's Bureau. PMID- 19310550 TI - Faculty of dental surgery: twenty-first anniversary celebrations, 17th-19th july 1968. PMID- 19310551 TI - The arms of the college. PMID- 19310552 TI - Faculty of dental surgery: dinner to commemorate the 21st anniversary. PMID- 19310553 TI - "Observables" at the royal college of surgeons: 53. The frankau cup. PMID- 19310554 TI - Observations of the blood flow in the inferior mesenteric arterial system, and the healing of colonic anastomoses. PMID- 19310555 TI - Faculty of Dental Surgery: Twenty-first Anniversary Celebrations. PMID- 19310556 TI - Christmas cards. PMID- 19310557 TI - The british club for surgery of the hand. PMID- 19310558 TI - Faculty of anaesthetists. PMID- 19310559 TI - Rheumatoid arthritis. PMID- 19310560 TI - Faculty of dental surgery. PMID- 19310561 TI - Holiday lectures for young people. PMID- 19310562 TI - Diploma ceremony. PMID- 19310563 TI - Register of fellows. PMID- 19310564 TI - Great teachers of surgery in the past. PMID- 19310565 TI - Joint committee for higher surgical training. PMID- 19310566 TI - The college tie. PMID- 19310567 TI - Commissioned officers in the medical services of the british army, 1661-1960. PMID- 19310568 TI - Hunterian festival. PMID- 19310569 TI - The basophil adenomas of the pituitary body. PMID- 19310570 TI - Measurement and precision in surgery. PMID- 19310571 TI - Courses in clinical surgery. PMID- 19310572 TI - Exhibitions. PMID- 19310573 TI - Institute of urology. PMID- 19310575 TI - Sir cecil wakeley, bt. PMID- 19310574 TI - Concourse. PMID- 19310576 TI - Cerebral circulation in anaesthesia. PMID- 19310578 TI - V Haematology related to surgery. PMID- 19310577 TI - Summary. PMID- 19310579 TI - The pathology, diagnosis and management of malignant tumours of the ovary. PMID- 19310580 TI - The management of renal calculi. PMID- 19310581 TI - Chairman'S comments. PMID- 19310582 TI - Chairman'S comments. PMID- 19310583 TI - Arterial stenoses. PMID- 19310584 TI - 'And one man in his time....'. PMID- 19310585 TI - 'Home is where one starts from...'. PMID- 19310587 TI - Some recollections of seventy years with the College. PMID- 19310586 TI - Publications. PMID- 19310588 TI - Court and council. PMID- 19310589 TI - The ceremonial mace. PMID- 19310590 TI - THE LIBRARY OF THE ROYAL COLLEGE OF SURGEONS OF ENGLAND: Books added to the Library, January-September 1972. PMID- 19310591 TI - St. George'S hospital and s.w. Metropolitan orthopaedic training scheme. PMID- 19310592 TI - ADVANCED SURGICAL COURSE: Suitable for F.R.C.S. Candidates Medical Education Centre, Whipps Cross Hospital, E.11 Mondays to Fridays 9 a.m. to 6.30 p.m. PMID- 19310594 TI - Hamilton bailey prize. PMID- 19310593 TI - Proposals for the future training of anaesthetists. PMID- 19310596 TI - ADVANCED SURGICAL COURSE: Suitable for F.R.C.S. Candidates Whipps Cross Hospital 1st-19th October 1973. PMID- 19310597 TI - County of hampshire medical Federation. PMID- 19310599 TI - First world congress on intensive care: patron: h.R.h. The duke of edinburgh, k.g., k.T. PMID- 19310598 TI - FINAL F.R.C.S. COURSE BROOK, GUY'S, AND LEWISHAM HOSPITALS: 10th-28th September 1973. PMID- 19310600 TI - British association of surgical oncology. PMID- 19310601 TI - British council for rehabilitation of the disabled: fifth international seminar and exhibition. PMID- 19310602 TI - Advanced surgical course: whipps cross hospital. PMID- 19310603 TI - Postgraduate surgery 1973-74 liverpool. PMID- 19310604 TI - The College and its younger fellows. PMID- 19310605 TI - Winston churchill memorial trust fellowships. PMID- 19310606 TI - Windsor and District Postgraduate Medical Centre: study days. PMID- 19310607 TI - St. George's Hospital and S.W. Metropolitan Orthopaedic Training Scheme. PMID- 19310608 TI - The Medical Commission on Accident Prevention: Convention on the Medical Aspects of Education in the Home and at School in the Prevention of Trauma, to be held at the Royal College of Surgeons of England on Wednesday 21st November 1973. PMID- 19310609 TI - Final f.R.C.s. Day release course: brook general hospital. PMID- 19310611 TI - Annals of the royal college of surgeons of England. PMID- 19310614 TI - The britain-Nepal medical trust. PMID- 19310615 TI - Letters from the past: From John Hunter to Edward Jenner. PMID- 19310622 TI - OPHTHALMOLOGICAL SOCIETY OF THE UNITED KINGDOM: Brief History of the Society. PMID- 19310617 TI - Letters from the past: From John Hunter to Edward Jenner. PMID- 19310626 TI - Letters from the past: From John Hunter to Edward Jenner. PMID- 19310629 TI - REPORT OF THE BOARD OF EXAMINERS FOR THE FELLOWSHIP IN DENTAL SURGERY, 17th JANUARY 1975. PMID- 19310628 TI - Research at the IBMS. 1974: Institute of Basic Medical Sciences, Royal College of Surgeons of England and British Postgraduate Medical Federation (University of London). PMID- 19310630 TI - Gifts to the college. PMID- 19310638 TI - From the president. PMID- 19310639 TI - Surgery for the aged - Introduction by the Chairman. PMID- 19310641 TI - Correction: A concise surgery of the acute abdomen. PMID- 19310643 TI - Research at the IBMS, 1975: Institute of Basic Medical Sciences, Royal College of Surgeons of England and British Postgraduate Medical Federation (University of London). PMID- 19310646 TI - BRITISH ASSOCIATION OF OTOLARYNGOLOGISTS: Excerpts from Annual Report of Council 1974/75. PMID- 19310645 TI - Norman Capener's 'Clasped hands'. PMID- 19310648 TI - Thoughts coming out of the ether. PMID- 19310650 TI - Sir Harry Platt and the IFSC: A 90th birthday tribute. PMID- 19310651 TI - The role of the professions in modern society. PMID- 19310652 TI - The rashid hospital, dubai. PMID- 19310653 TI - Fellows of fifty years standing. PMID- 19310655 TI - Norman capener travelling fellowship. PMID- 19310654 TI - Secretary'S tour of australia. PMID- 19310656 TI - REPORT OF THE BOARD OF EXAMINERS FOR THE FELLOWSHIP IN DENTAL SURGERY, 25th June 1976. PMID- 19310657 TI - BRITISH ASSOCIATION OF OTOLARYNGOLOGISTS: Excerpts from the Honorary Secretary's Newsletter, June 1976. PMID- 19310659 TI - CONSULTATIVE DOCUMENT ON PRIORITIES FOR HEALTH AND PERSONAL SOCIAL SERVICES IN ENGLAND: Evidence from the Royal College of Surgeons of England. PMID- 19310660 TI - Sir harry platt. PMID- 19310661 TI - REPORT OF THE BOARD OF EXAMINERS FOR THE FELLOWSHIP IN THE FACULTY OF ANAESTHETISTS 23rd JULY 1976. PMID- 19310663 TI - Everest: recollections and reflections. PMID- 19310664 TI - PROVISION OF DONOR ORGANS AND TISSUES: Document prepared by a working party of the Royal College of Surgeons of England. PMID- 19310665 TI - DIAGNOSIS OF BRAIN DEATH: Statement issued by the Honorary Secretary of the Conference of Medical Royal Colleges and their Faculties in the United Kingdom on 11th October 1976. PMID- 19310667 TI - REPORT OF THE COURT OF EXAMINERS, 24th NOVEMBER 1976. PMID- 19310668 TI - Addresses wanted. PMID- 19310670 TI - Academic posts in relation to proposals of the joint committee for higher training in dentistry. PMID- 19310671 TI - SINGAPORE, AUGUST 1976: Examiners in the Final MMed (Surgery) Examination. PMID- 19310672 TI - REPORT OF THE BOARD OF EXAMINERS FOR THE FELLOWSHIP IN THE FACULTY OF ANAESTHETISTS, 11th February 1977. PMID- 19310673 TI - REPORT OF THE BOARD OF EXAMINERS FOR THE FELLOWSHIP IN DENTAL SURGERY, 14th JANUARY 1977. PMID- 19310676 TI - Hunterian festival 1978. PMID- 19310678 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the first scientific meeting. PMID- 19310679 TI - Pancreatic Society of Great Britain and Ireland: Abstracts of papers presented at the second Annual Meeting. PMID- 19310681 TI - The College's new Charter. PMID- 19310680 TI - Commemorative bust of John Hunter. PMID- 19310683 TI - The festival. PMID- 19310687 TI - Royal society conversazione, 1978: exhibit by department of dental science. PMID- 19310684 TI - Correction: appointment of fellows to consultant and similar posts. PMID- 19310690 TI - A smith and nephew fellowship. PMID- 19310691 TI - REPORT OF THE BOARD OF EXAMINERS FOR THE FELLOWSHIP IN THE FACULTY OF ANAESTHETISTS, 21st JULY 1978. PMID- 19310692 TI - Correction: appointments of fellows to consultant and similar posts. PMID- 19310696 TI - Uniform requirements for manuscripts submitted to biomedical journals. PMID- 19310698 TI - Notice to contributors. PMID- 19310700 TI - Notice to contributors. PMID- 19310701 TI - Local anaesthesia for herniorraphy. PMID- 19310702 TI - Kutler repair for amputated fingertip. PMID- 19310703 TI - Gladys Sandes FRCS England 1897-1968. PMID- 19310705 TI - Female urinary incontinence. PMID- 19310707 TI - Transabdominal bilateral adrenalectomy for metastatic breast carcinoma. PMID- 19310706 TI - Contact cholangiography. PMID- 19310708 TI - Transabdominal bilateral adrenalectomy for metastatic breast carcinoma. PMID- 19310709 TI - Transabdominal bilateral adrenalectomy for metastatic breast carcinoma. PMID- 19310710 TI - Transabdominal bilateral adrenalectomy for metastatic breast carcinoma. PMID- 19310711 TI - Transabdominal bilateral adrenalectomy for metastatic breast carcinoma. PMID- 19310712 TI - Evaluation of dextran with local anaesthesia for herniorraphy. PMID- 19310713 TI - Evaluation of dextran with local anaesthesia for herniorraphy. PMID- 19310714 TI - Tinel sign in brachial plexus lesions. PMID- 19310716 TI - Courses in surgery. PMID- 19310718 TI - Local anaesthesia for repair of groin hernias. PMID- 19310719 TI - Respiratory obstruction in thyroid surgery. PMID- 19310720 TI - Team approach in the management of oral cancer. PMID- 19310721 TI - Team approach in the management of oral cancer. PMID- 19310722 TI - Sutureless skin closure. PMID- 19310725 TI - The present role of the Fellowship. PMID- 19310724 TI - A voice from the past. PMID- 19310726 TI - Local anaesthesia for repair of groin hernia. PMID- 19310727 TI - Local anaesthesia for repair of groin hernia. PMID- 19310728 TI - Eastman dental hospital jubilee, 1930-1980. PMID- 19310729 TI - Diagnosis of primary hyperparathyroidism. PMID- 19310731 TI - In praise of theatre users committees. PMID- 19310733 TI - Fresh blood for massive transfusions. PMID- 19310734 TI - Prostatectomy at a district general hospital. PMID- 19310735 TI - Fresh blood for massive transfusions. PMID- 19310736 TI - Cross-leg flap repairs. PMID- 19310738 TI - The surgical senior lecturer. PMID- 19310737 TI - The surgical senior lecturer. PMID- 19310739 TI - Cross-leg flap repairs. PMID- 19310741 TI - Drainage after cholecystectomy. PMID- 19310740 TI - The surgical senior lecturer. PMID- 19310743 TI - Surgery in developing countries. PMID- 19310744 TI - The senior surgical lecturer. PMID- 19310745 TI - The senior surgical lecturer. PMID- 19310746 TI - The senior surgical lecturer. PMID- 19310748 TI - British association of clinical anatomists, summer meeting, 1980. PMID- 19310750 TI - Preoperative assessment for outpatient anaesthesia. PMID- 19310749 TI - Surgery in developing countries. PMID- 19310751 TI - Preoperative assessment for outpatient anaesthesia. PMID- 19310752 TI - Perioperative steroid supplements. PMID- 19310753 TI - Courses in surgery. PMID- 19310754 TI - Aphorisms in cancer surgery. PMID- 19310756 TI - Training and careers. PMID- 19310757 TI - Aphorisms in cancer surgery. PMID- 19310760 TI - Higher specialist training. PMID- 19310759 TI - Higher specialist training. PMID- 19310761 TI - Higher specialist training. PMID- 19310764 TI - Anglo-French meeting on venous disorders, Le Touquet, 8th-9th May 1981. PMID- 19310762 TI - Wrong chi-squared again. PMID- 19310765 TI - British association of clinical anatomists. Annual general meeting, 1981. PMID- 19310767 TI - Dextran in local anaesthesia. PMID- 19310766 TI - Colorectal liver metastases. PMID- 19310768 TI - Rehabilitation of the hand in tetraplegics. PMID- 19310769 TI - History and evolution of surgical instruments. PMID- 19310770 TI - Preparation of a Mastership thesis. PMID- 19310771 TI - Gore-tex. PMID- 19310772 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310773 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310776 TI - Editorial. PMID- 19310775 TI - ROYAL COLLEGE OF SURGEONS OF ENGLAND: Report on Surgical Manpower and the Career Structure. PMID- 19310777 TI - Pancreatic society of great britain and ireland: annual meeting, 1980: second scientific session. AB - With the increase in the number of diabetic patients in renal failure new attempts are being made to improve both the results of cadaveric transplantation and the quality of rehabilitation. A combined approach of renal and segmental pancreatic transplantation has been made in patients with major diabetic complications. We here report the early Cambridge experience using this management. PMID- 19310778 TI - Pancreatic Society of Great Britain and Ireland: Annual Meeting, 1980: Role of pH dependent bile acid precipitation in fat maldigestion due to pancreatic steatorrhoea. Preliminary communication. AB - In pancreatic steatorrhoea due to cystic fibrosis (CF) a major proportion of the meal enters the jejunum below the critical pH of 5, causing bile acid (BA) precipitation and limiting aqueous solubilisation of lipid. Treatment with pancreatin alone results only in a small increase in lipolysis as the lipase is largely inactivated; aqueous lipid solubilisation is not improved as BA precipitation remains a limiting factor. Treatment with cimetidine alone, by reducing BA precipitation, improves lipid solubilisation without improving lipolysis. Treatment with cimetidine and pancreatin reduces pancreatic lipase inactivation and BA precipitation leading to the greatest improvement in lipid solubilisation. PMID- 19310779 TI - The 50th anniversary of the royal college of physicians and surgeons of Canada. PMID- 19310780 TI - The research backwater. PMID- 19310781 TI - Higher specialist training. PMID- 19310782 TI - Dial-a-Flo: a warning. PMID- 19310783 TI - Exclusion pyloroplasty. PMID- 19310784 TI - Rehabilitation of the hand in tetraplegics. PMID- 19310785 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310786 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310787 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310788 TI - Operative cholangiography. PMID- 19310790 TI - Surgery in developing countries. PMID- 19310789 TI - Salivary gland tumour panel. PMID- 19310792 TI - Dial-a-Flo: an explanation. PMID- 19310793 TI - Is a mask necessary in the operating theatre? PMID- 19310794 TI - Is a mask necessary in the operating theatre? PMID- 19310795 TI - Chemical lumbar sympathectomy with radiological assessment. PMID- 19310796 TI - Chemical lumbar sympathectomy with radiological assessment. PMID- 19310798 TI - Chemical lumbar sympathectomy with radiological assessment. PMID- 19310797 TI - Chemical lumbar sympathectomy with radiological assessment. PMID- 19310799 TI - Early diagnosis of carcinoma of the mouth. PMID- 19310800 TI - Early diagnosis of carcinoma of the mouth. PMID- 19310801 TI - Early diagnosis of carcinoma of the mouth. PMID- 19310802 TI - Aphorisms in cancer surgery. PMID- 19310803 TI - Advantages of stay sutures with tracheostomy. PMID- 19310805 TI - The westerside story. PMID- 19310806 TI - British association of clinical anatomists: summer meeting 1981. PMID- 19310807 TI - Aspects of audit. PMID- 19310808 TI - Aspects of audit. PMID- 19310810 TI - Advantages of stay sutures with tracheostomy. PMID- 19310809 TI - Surgical manpower and career structure. PMID- 19310811 TI - Early closure of transverse loop colostomies. PMID- 19310812 TI - Cholangiography or choledochoscopy? PMID- 19310814 TI - Higher specialist training. PMID- 19310813 TI - Early closure of transverse loop colostomies. PMID- 19310815 TI - Chest injuries in a district general hospital. PMID- 19310816 TI - Is a mask necessary in the operating theatre? PMID- 19310818 TI - Visits to India and pakistan. PMID- 19310819 TI - On the way to the theatre. PMID- 19310820 TI - The short report. PMID- 19310821 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310822 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310824 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310823 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310825 TI - Endoscopic lower ureterectomy. PMID- 19310826 TI - Appendicectomy in children: transverse pararectal approach. PMID- 19310827 TI - Cholecystokinin in diagnosis of biliary pain. PMID- 19310828 TI - Cholecystokinin in diagnosis of biliary pain. PMID- 19310829 TI - Cholecystokinin in diagnosis of biliary pain. PMID- 19310830 TI - Chest injuries in a district general hospital. PMID- 19310831 TI - Chest injuries in a district general hospital. PMID- 19310833 TI - Dr frank stansfield. PMID- 19310832 TI - Appendicitis in external herniae. PMID- 19310834 TI - Drainage after cholecystectomy. PMID- 19310836 TI - Courses in surgery. PMID- 19310835 TI - Drainage after cholecystectomy. PMID- 19310838 TI - New medical school in Penang. PMID- 19310839 TI - Anaesthesia for microsurgery of the larynx. PMID- 19310840 TI - Anaesthesia for microsurgery of the larynx. PMID- 19310841 TI - Anaesthesia for microsurgery of the larynx. PMID- 19310842 TI - Venous thromboembolism. PMID- 19310843 TI - Drainage after cholecystectomy. PMID- 19310844 TI - Prophylactic use of hypochlorite. PMID- 19310845 TI - Treatment of respiratory failure after chest injury. PMID- 19310847 TI - The short report. PMID- 19310846 TI - Cholangiography or choledochoscopy? PMID- 19310850 TI - Pancreatic Society of Great Britain and Ireland: Sixth Annual Meeting, 1981: POSTER PRESENTATIONS. PMID- 19310848 TI - Admissions to court of patrons. PMID- 19310851 TI - The role of peritoneal lavage in the prediction and treatment of severe acute pancreatitis: Commentary. PMID- 19310852 TI - Formal duodenal fistula. PMID- 19310853 TI - Formal duodenal fistula. PMID- 19310854 TI - Toxic dilatation and perforation in inflammatory bowel disease. PMID- 19310855 TI - Facial expression in acute appendicitis. PMID- 19310857 TI - Editorial. PMID- 19310858 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Annual General Meeting, 1982. PMID- 19310859 TI - Thyroidectomy: is Lugol's iodine necessary? PMID- 19310860 TI - Oestrogen receptor analysis in early cancer of the breast. PMID- 19310861 TI - Thyroidectomy: is Lugol's iodine necessary? PMID- 19310862 TI - Thyroidectomy: is Lugol's iodine necessary? PMID- 19310863 TI - Radical Surgery for pilonidal sinus. PMID- 19310864 TI - Radical Surgery for pilonidal sinus. PMID- 19310865 TI - Thyroidectomy: is Lugol's iodine necessary? PMID- 19310866 TI - Facial expression in acute appendicitis. PMID- 19310867 TI - Facial expression in acute appendicitis. PMID- 19310868 TI - Facial expression in acute appendicitis. PMID- 19310869 TI - A randomised controlled trial to compare local with general anaesthesia for short stay inguinal hernia repair. PMID- 19310870 TI - Exteriorisation resection of the colon. PMID- 19310871 TI - Treatment of respiratory failure after chest injury. PMID- 19310872 TI - Methods for relief of postoperative pain. PMID- 19310873 TI - Methods for relief of postoperative pain. PMID- 19310874 TI - Radical surgery for pilonidal sinus. PMID- 19310875 TI - Methods for relief of postoperative pain. PMID- 19310877 TI - Methods for relief of postoperative pain. PMID- 19310876 TI - Methods for relief of postoperative pain. PMID- 19310878 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Summer Meeting, 1982. PMID- 19310880 TI - Toxic dilatation and perforation in inflammatory bowel disease. PMID- 19310881 TI - How do surgeons treat haemarrhoids? PMID- 19310882 TI - An open comparison between routine and self-administered postoperative pain relief. PMID- 19310883 TI - Subcutaneous bupivacaine for postoperative analgesia after herniorrhaphy. PMID- 19310884 TI - Plain bupivacaine 0.5%: a preliminary evaluation as a spinal anaesthetic agent. PMID- 19310885 TI - Plain bupivacaine 0.5%: a preliminary evaluation as a spinal anaesthetic agent. PMID- 19310886 TI - Are systemic prophylactic antibiotics necessary for burns? PMID- 19310887 TI - Bleeding and cupping. PMID- 19310888 TI - A common bile duct sound: an aid to sphincterotomy. PMID- 19310889 TI - A common bile duct sound: an aid to sphincterotomy. PMID- 19310890 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Annual General Meeting, 1983. AB - The Annual General Meeting of the British Association of Clinical Anatomists for 1983 was held at the Royal College of Surgeons of England on 14th January 1983. The following are abstracts of the papers presented. PMID- 19310891 TI - Plain bupivacaine 0.5%: a preliminary evaluation as a spinal anaesthetic agent. PMID- 19310892 TI - Prolonged access to the venous system using the Hickman right atrial catheter. PMID- 19310893 TI - Prolonged access to the venous system using the Hickman right atrial catheter. PMID- 19310894 TI - Some changing aspects of primary hyperparathyroidism. PMID- 19310895 TI - Are systemic prophylactic antibiotics necessary for burns? PMID- 19310896 TI - Op-Site skin closure. PMID- 19310897 TI - A modified Gritti-Stokes amputation. PMID- 19310898 TI - Awareness during anaesthesia: a review. PMID- 19310899 TI - A modified Gritti-Stokes amputation. PMID- 19310900 TI - Awareness during anaesthesia: a review. PMID- 19310901 TI - A simple stoma wafer cutter. PMID- 19310902 TI - Simple renal cysts in children. PMID- 19310903 TI - A study of colovesical fistulae in a district hospital. PMID- 19310905 TI - A simple stoma wafer cutter. PMID- 19310904 TI - The fate of the below knee amputee. PMID- 19310906 TI - Haemarrhoids treated by cryotherapy: a critical analysis. PMID- 19310907 TI - Campbell De Morgan and his spots. PMID- 19310908 TI - A useful eye sign in the apparently unconscious patient. PMID- 19310909 TI - A combination of subcuticular suture and sterile Micropore tape compared with conventional interrupted sutures for skin closure. PMID- 19310910 TI - The direct perfusion of surgical wounds with local anaesthetic solution: an approach to post-operative pain? PMID- 19310911 TI - The microflora of the oesophagus. PMID- 19310912 TI - Haemorrhoids treated by cryotherapy. PMID- 19310913 TI - The fate of the below knee amputee. PMID- 19310915 TI - Problems of the staffing of intensive care units in British district general hospitals. PMID- 19310914 TI - Problems of the staffing of intensive care units in British district general hospitals. PMID- 19310916 TI - A useful eye sign in the apparently unconscious patient. PMID- 19310917 TI - A useful eye sign in the apparently unconscious patient. PMID- 19310918 TI - Campbell de Morgan and his spots. PMID- 19310919 TI - El Zahrawi (936-1013 AD), the father of operative surgery. PMID- 19310920 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Summer Meeting, 1983. PMID- 19310921 TI - Transduodenal per-ampullary approach to common bile duct calculi. PMID- 19310922 TI - A common bile duct sound: an aid to sphincterotomy. PMID- 19310924 TI - The fate of the below knee amputee. PMID- 19310923 TI - A simple, safe, surgical technique for closing the persistent ductus arteriosus in the preterm neonate. PMID- 19310925 TI - Surgical wound management with adhesive polyurethane membrane: a preferred method for routine usage. PMID- 19310927 TI - Surgical wound management with adhesive polyurethane membrane: a preferred method for routine usage. PMID- 19310926 TI - Right hemicolectomy: use of GIA stapler. PMID- 19310928 TI - A useful eye sign in the apparently unconscious patient. PMID- 19310929 TI - Emergency surgery for stab wounds to the heart. PMID- 19310930 TI - A useful eye sign in the apparently unconscious patient. PMID- 19310931 TI - Unoperated abdominal aortic aneurysms: presentation and natural history. PMID- 19310932 TI - Emergency surgery for stab wounds to the heart. PMID- 19310934 TI - The restoration of laryngeal competence by polytetrafluorethylene paste after oesophageal surgery. PMID- 19310933 TI - Removal of common bile duct stones after emergency drainage of the gallbladder. PMID- 19310935 TI - Undergraduate students' experience in 'peripheral' and 'teaching' hospitals compared. PMID- 19310936 TI - Regional anaesthesia in elective hand surgery. PMID- 19310938 TI - Regional anaesthesia in elective hand surgery. PMID- 19310937 TI - Regional anaesthesia in elective hand surgery. PMID- 19310939 TI - Surgical wound management with adhesive polyurethane membrane. PMID- 19310940 TI - Long-term efficacy of surgical cordotomy in intractable non-malignant pain. PMID- 19310941 TI - A comparison of skin grafting and healing by granulation, following axillary excision for hidradenitis suppurativa. PMID- 19310942 TI - Emergency surgery for stab wound to the heart. PMID- 19310943 TI - Transtracheal ventilation in oral surgery. PMID- 19310945 TI - Undergraduate students' experience in 'peripheral' and 'teaching' hospitals compared. PMID- 19310944 TI - The use of pedicled transplants of sigmoid colon or other parts of the intestinal tract for vaginal reconstruction. PMID- 19310947 TI - The use of pedicled transplants of sigmoid colon or other parts of the intestinal tract for vaginal reconstruction. PMID- 19310946 TI - Airway management in the transfer of the unconscious patient. PMID- 19310948 TI - Towards an efficient retractor handle: an ergonomic study. PMID- 19310949 TI - Minor burns in children; inpatient versus outpatient treatment. PMID- 19310950 TI - A proposed simple classification of musculoskeletal pain. PMID- 19310951 TI - A proposed simple classification of musculoskeletal pain. PMID- 19310952 TI - Delayed rupture of the spleen can masquerade as appendicitis. PMID- 19310953 TI - The arterial tourniquet. PMID- 19310954 TI - A proposed simple classification of musculoskeletal pain. PMID- 19310955 TI - Sir benjamin brodie, bt (1783-1862). PMID- 19310956 TI - Editorial. PMID- 19310957 TI - The direct perfusion of surgical wounds with local anaesthetic solution: an approach to postoperative pain? PMID- 19310958 TI - Epiglottopexy: a new surgical technique to prevent intractable aspiration. PMID- 19310960 TI - Identification of accessory bile ducts at cholecystectomy. PMID- 19310959 TI - The fate of the below-knee amputee. PMID- 19310961 TI - Identification of accessory bile ducts at cholecystectomy. PMID- 19310962 TI - Postoperative care: the role of the High Dependency Unit. PMID- 19310964 TI - The place of antibiotics in preventing wound infection. PMID- 19310963 TI - The use of pedicled transplants of sigmoid colon or other parts of the intestinal tract for vaginal reconstruction. PMID- 19310965 TI - Proposed simple classification of Musculo-skeletal pain. PMID- 19310967 TI - The role of the Intensive Care Unit in a District Hospital. PMID- 19310966 TI - Peritoneal drainage following cholecystectomy: a controlled trial. PMID- 19310968 TI - The prevention of wound infection after head and neck resections. PMID- 19310969 TI - Complete rectal prolapse: repair by a simple technique. PMID- 19310970 TI - Major vascular injuries associated with orthopaedic injuries to the lower limb. PMID- 19310971 TI - Major vascular injuries associated with orthopaedic injuries to the lower limb. PMID- 19310972 TI - The organisation and evaluation of an open-access dysphagia clinic. PMID- 19310973 TI - Peritoneal drainage following cholecystectomy: a controlled trial. PMID- 19310974 TI - The management of acute colonic diverticulitis with suppurative peritonitis. PMID- 19310975 TI - The management of acute colonic diverticulitis with suppurative peritonitis. PMID- 19310976 TI - The management of acute colonic diverticulitis with suppurative peritonitis. PMID- 19310978 TI - Laser modalities in neurosurgery. PMID- 19310977 TI - The appendix stump: should it be invaginated? PMID- 19310979 TI - The appendix stump: should it be invaginated? PMID- 19310980 TI - The appendix stump: should it be invaginated? PMID- 19310981 TI - The appendix stump: should it be invaginated? PMID- 19310982 TI - Balloon tamponade in the management of bleeding oesophageal varices. PMID- 19310984 TI - Surgical emergencies and manpower. PMID- 19310983 TI - Surgical emergencies and manpower. PMID- 19310985 TI - The best back to manipulate? PMID- 19310986 TI - Surgical wound management with adhesive polyurethane membraine. PMID- 19310987 TI - Surgical research-curiosity and luck: personal observations and prejudices. PMID- 19310988 TI - Thomas Dunhill 1876 1957 Pioneer of thyroid surgery. PMID- 19310989 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Annual General Meeting, 1984. PMID- 19310990 TI - The appendix stump: should it be invaginated? PMID- 19310991 TI - The place of antibiotics in preventing wound infection after clean operations in an Indian hospital. PMID- 19310992 TI - Peritoneal drainage following cholecystecomy: a controlled trial. PMID- 19310993 TI - Transduodenal exploration of the common bile duct in a district general hospital. PMID- 19310994 TI - New perspectives in the management of severe craniofacial deformity. PMID- 19310996 TI - Acute pseudo-obstruction of the colon. PMID- 19310995 TI - Comparison of early and late closure of transverse loop colostomies. PMID- 19310997 TI - Surgical wound management with adhesive polyurethane membrane. PMID- 19311000 TI - Evaluation of anti-emetics in association with intrathecal diamorphine. PMID- 19310998 TI - Subcutaneous left colon reconstruction following pharyngo-laryngectomy. PMID- 19311001 TI - Motility changes associated with large bowel obstruction and its surgical relief. PMID- 19311002 TI - Inguinal hernia repair using local anaesthesia. PMID- 19311004 TI - Intravital methylene blue staining of parathyroid glands and tumours. PMID- 19311003 TI - Replacement of the pharynx after pharyngolaryngectomy. PMID- 19311005 TI - A new tumour marker tested in 98 patients with bladder carcinoma. PMID- 19311006 TI - The Biethium Bridge-an advance in stoma care. PMID- 19311007 TI - Trial of the use of masks in the gynaecological operating theatre. PMID- 19311008 TI - The Biethium Bridge-an advance in stoma care. PMID- 19311009 TI - Orthopaedic Audit: one year's experience in a District General Hospital. PMID- 19311010 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Summer Meeting, July 1984. PMID- 19311011 TI - Surgical emergencies and manpower. PMID- 19311012 TI - Surgical wound management with adhesive polyurethane membrane. PMID- 19311013 TI - New perspectives in the management of severe craniofacial deformity. PMID- 19311014 TI - Inguinal hernia: cremasteric muscle sling operation. PMID- 19311015 TI - Motility changes associated with large bowel obstruction and its surgical relief. PMID- 19311016 TI - Anaesthesia for transcervical thymectomy in myasthenia gravis. PMID- 19311017 TI - Anaesthesia for transcervical thymectomy in myasthenia gravis. PMID- 19311018 TI - The anaesthetic management of the Eisenmenger syndrome. PMID- 19311019 TI - Inguinal hernia repair using local anaesthesia. PMID- 19311020 TI - The provision of junior anaesthetic staff for the Intensive Care Unit of a District General Hospital: a workable solution? PMID- 19311021 TI - A new tumour marker tested in 98 patients with bladder carcinoma. PMID- 19311022 TI - Ketamine: effect of literacy on emergence phenomena. PMID- 19311023 TI - Trial of the use of masks in the gynaecological operating theatre. PMID- 19311025 TI - Use of a snugging ligature for securing a cholangiogram catheter into the cystic duct. PMID- 19311024 TI - Use of a snugging ligature for securing a cholangiogram catheter into the cystic duct. PMID- 19311026 TI - Recent experience with arterial embolism of the limb in a vascular unit. PMID- 19311028 TI - Cervical thymectomy in the treatment of myasthenia gravis. PMID- 19311027 TI - Subcutaneous left colon reconstruction following, pharyngo-laryngectomy. PMID- 19311029 TI - Experimental deep venous thrombogenesis by a non-invasive method. PMID- 19311030 TI - Biopsy procedures, primary wide excisional surgery and long-term prognosis in primary clinical stage I invasive cutaneous malignant melanoma. PMID- 19311031 TI - Conservative management of appendix mass in children. PMID- 19311032 TI - Penetrating injuries of the neck. PMID- 19311034 TI - An assessment of operative choledochoscopy-A worthwhile procedure or not? Choledochoscopy? Post-exploratory fluorocholangiography? Or both? PMID- 19311033 TI - Peroperative cholangiography and post-cholecystectomy strictures. PMID- 19311035 TI - The role of percutaneous transluminal angioplasty for atherosclerotic disease of the lower extremities. PMID- 19311036 TI - Interrupted subcuticular polyglactin sutures for abdominal wounds. PMID- 19311038 TI - Read this. PMID- 19311037 TI - Surgical wound management with adhesive polyurethane membrane. PMID- 19311039 TI - Terence p N jenkins. PMID- 19311040 TI - Surgical emergencies and manpower. PMID- 19311041 TI - Anaesthesia for transcervical thymectomy in myasthenia gravis. PMID- 19311042 TI - An assessment of operative choledochoscopy-a worthwhile procedure or not? PMID- 19311043 TI - Peroperative cholangiography and postcholecystectomy biliary strictures. PMID- 19311044 TI - Preserved dura and pericardium for closure of large abdominal wall and diaphragmatic defects in children. PMID- 19311046 TI - An assessment of operative choledochoscopy-a worthwhile procedure or not? PMID- 19311045 TI - Peroperative cholangiography and postcholecystectomy biliary strictures. PMID- 19311047 TI - Use of a snugging ligature for securing a cholangiogram catheter into the cystic duct. PMID- 19311048 TI - Silastic foam dressings: appraisal. PMID- 19311049 TI - Changing knives a wasteful and unnecessary ritual. PMID- 19311050 TI - Changing knives a wasteful and unnecessary ritual. PMID- 19311051 TI - Does mass closure of midline laparotomies stand the test of time? PMID- 19311052 TI - Post-tonsillectomy secondary haemorrhage. PMID- 19311053 TI - Intraoral carcinoma in Nigeria-a review of 137 cases. PMID- 19311054 TI - The role of anaesthetic management in free flap transfer. PMID- 19311055 TI - Cuthbert E Dukes OBE MD MSc FRCS FRCPath DPH -1890-1977. PMID- 19311056 TI - Experience with surgical implantation of catheters for continuous ambulatory peritoneal dialysis. PMID- 19311057 TI - Changing knives-a wasteful and unnecessary ritual. PMID- 19311058 TI - Girth measurement is not a reliable investigation for the detection of intra abdominal fluid. PMID- 19311060 TI - An anatomical curiosity. PMID- 19311059 TI - The association between lowest palpable pulse and wound healing in below knee amputation. PMID- 19311061 TI - Caecal volvulus: a method of management. PMID- 19311062 TI - The importance of the lymphatic system. PMID- 19311063 TI - Editorial. PMID- 19311064 TI - Inguinal Hernia Repair using Local Anaesthesia. PMID- 19311065 TI - Inguinal Hernia Repair using Local Anaesthesia. PMID- 19311067 TI - Changing knives a wasteful and unnecessary routine. PMID- 19311066 TI - Vertical banded gastroplasty: operation for morbid obesity. PMID- 19311068 TI - An anatomical curiosity. PMID- 19311069 TI - The use of a microcomputer for in-patients audit in an orthopaedic department. PMID- 19311070 TI - An anatomical curiosity. PMID- 19311071 TI - Abdominal wound closure: A controlled trial of polyamide and polydioxanone suture (PDS). PMID- 19311073 TI - Breast reconstruction after failed conservation. PMID- 19311072 TI - Staples for wound closure: a controlled trial. PMID- 19311074 TI - Staples for wound closure: a controlled trial. PMID- 19311075 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Annual General Meeting on 11th January 1985 at The Queen Elizabeth Postgraduate Centre, University of Birmingham. PMID- 19311076 TI - Peroperative cholangiography and postcholecystectomy biliary strictures. PMID- 19311077 TI - Surgical wound management with adhesive polyurethane membrane. PMID- 19311078 TI - Penetrating injuries of the neck. PMID- 19311079 TI - Five year experience using PTFE vascular grafts for lower limb ischaemia. PMID- 19311080 TI - Complications of T-tube drainage of the common bile duct. PMID- 19311081 TI - Continuous subcutaneous pethidine for routine postoperative analgesia. PMID- 19311082 TI - Operative choledochoscopy in common bile duct surgery. PMID- 19311083 TI - Breast reconstruction after failed conservation. PMID- 19311084 TI - A non-stick dressing suitable for anal wounds. PMID- 19311085 TI - Critical comment. PMID- 19311086 TI - Abdominocervical oesophagectomy in the elderly. PMID- 19311087 TI - Caecal volvulus: A method of management. PMID- 19311088 TI - The changing trend in prostatectomy in a district general hospital. PMID- 19311089 TI - Endoscopic sphincterotomy as an alternative to common bile duct exploration and T tube drainage. PMID- 19311090 TI - An aid to the assessment of bowel preparation prior to colonic resection. PMID- 19311091 TI - The changing trend in prostatectomy in a district general hospital. PMID- 19311092 TI - Continuous subcutaneous pethidine for routine postoperative analgesia. PMID- 19311093 TI - Breast conservation after failed conservation. PMID- 19311094 TI - Breast conservation after failed conservation. PMID- 19311095 TI - Biopsy procedures, primary wide excisional surgery and long term prognosis in primary clinical stage I invasive cutaneous malignant melanoma. PMID- 19311096 TI - Audit on complicated diverticular disease. PMID- 19311097 TI - Editorial. PMID- 19311098 TI - Read this. PMID- 19311100 TI - Use of the rhomboid flap in pilonidal sinus. PMID- 19311099 TI - A prospective randomised controlled trial of mezlocillin versus metilmicin in biliary surgery. PMID- 19311101 TI - The influence of microscopic disease at the margin of resection on recurrence rates in Crohn's disease. PMID- 19311103 TI - Is a drip necessary for a cholecystectomy? PMID- 19311102 TI - Plastic surgery in a rural tropical hospital: spectrum and implications. PMID- 19311104 TI - Breast reconstruction after failed conservation. PMID- 19311106 TI - Prediction of incisional hernia using radio-opaque markers. PMID- 19311105 TI - The use of laparoscopy in the management of right iliac fossa pain. PMID- 19311107 TI - Vertical banded gastroplasty: operation for morbid obesity. PMID- 19311108 TI - Breast reconstruction after failed conservation. PMID- 19311109 TI - Is a drip necessary for a cholecystectomy? PMID- 19311111 TI - Delorme's operation: the first choice in complete rectal prolapse? PMID- 19311110 TI - A method for identification of vertebral level. PMID- 19311112 TI - Use of the rhomboid flap in pilonidal sinus. PMID- 19311113 TI - Endoscopic thoracic sympathectomy in the treatment of upper limb hyperhydrosis. PMID- 19311114 TI - Morbidity study of submandibular gland excision. PMID- 19311115 TI - Endoscopic papillotomy without cholecystectomy for bile duct stones: Assessor's Comments. PMID- 19311116 TI - Below knee femoropopliteal bypass in severe ischaemia: results using EXS Dacron and human umbilical vein: Assessor's Comments. PMID- 19311117 TI - Gastrojejunostomy: A simple method of treatment of gastric volvulus. PMID- 19311119 TI - A review of 155 extra-anatomic bypass grafts. PMID- 19311118 TI - Complications of T-tube drainage of the common bile duct. PMID- 19311120 TI - Bupivicaine squirting. PMID- 19311121 TI - Is a drip necessary for cholecsytectomy? PMID- 19311122 TI - Primary suture or T-tube drainage after choledochotomy. PMID- 19311123 TI - Transcutaneous electrical nerve stimulation following appendicectomy: the placebo effect. PMID- 19311124 TI - Transcutaneous electrical nerve stimulation following appendicectomy: the placebo effect. PMID- 19311125 TI - Mortality in patients with obstructing colorectal cancer. PMID- 19311126 TI - Breast reconstruction after failed conservation. PMID- 19311127 TI - Breast reconstruction after failed conservation. PMID- 19311128 TI - Read this. PMID- 19311129 TI - The surgical management of pancreatic abscess: Assessors Comments. PMID- 19311130 TI - Perioperative complications of in-situ vein bypass graft. PMID- 19311131 TI - Breast reconstruction after failed conservation. PMID- 19311132 TI - Perioperative complications of in-situ vein bypass graft. PMID- 19311133 TI - Duodenal ulcer perforation: the effect of H(2) antagonists? PMID- 19311134 TI - Duodenal ulcer perforation: the effect of H(2) antagonists? PMID- 19311135 TI - Haemorrhage and perforation of gastrointestinal neoplasms during chemotherapy. PMID- 19311136 TI - Transcutaneous electrical nerve stimulation following appendicectomy: the placebo effect. PMID- 19311138 TI - Autotransfusion, an experience of seventy six cases. PMID- 19311137 TI - Transcutaneous electrical nerve stimulation following appendicectomy: the placebo effect. PMID- 19311139 TI - Below knee femoropopliteal bypass in severe ischaemia: results using EXS Dacron and human umbilical vein. PMID- 19311140 TI - Method of one-way surgical drainage for the inexpensive accurate measurement of abdominal secretions. PMID- 19311141 TI - A prospective randomised comparison of healing in Gritti-Stokes and through-knee amputations. PMID- 19311142 TI - Bupiv"i"caine squirting. PMID- 19311144 TI - A nutritional team is not essential for optimal parenteral nutrition therapy. PMID- 19311143 TI - A nutritional team is not essential for optimal parenteral nutrition therapy. PMID- 19311145 TI - Pitfalls in the assessment of dysphagia by fibreoptic oesophagogastroscopy. PMID- 19311147 TI - Breast reconstruction after failed conservation. PMID- 19311146 TI - Auditing perioperative mortality: Assessor's Comments. PMID- 19311148 TI - Breast reconstruction after failed conservation. PMID- 19311149 TI - A prospective randomised comparison of healing in Gritti-Stokes and through-knee amputations. PMID- 19311150 TI - Bupivacaine 'squirting'. PMID- 19311151 TI - Awareness during bronchoscopy. PMID- 19311153 TI - Polydioxanone suture in the gastrointestinal tract. PMID- 19311152 TI - Upper urinary tract stone disease: the changing management in a district general hospital. PMID- 19311154 TI - The surgical treatment of a pharyngeal pouch: inversion or excision? PMID- 19311155 TI - The surgical treatment of a pharyngeal pouch inversion or excision? PMID- 19311157 TI - Read this. PMID- 19311156 TI - Awareness during bronchoscopy. PMID- 19311158 TI - Terence john millin-1903-80. PMID- 19311160 TI - Colostomy or ileostomy after colorectal anastomosis?: a randomised trial. PMID- 19311159 TI - A review of 155 extra-anatomic bypass grafts. PMID- 19311161 TI - Overseas doctors training scheme. PMID- 19311162 TI - Missing chi again. PMID- 19311163 TI - Pitfalls in the assessment of dysphagia by fibreoptic oesophagogastroscopy. PMID- 19311165 TI - Recurrence rates following local anaesthetic day case inguinal hernia repair by junior surgeons in a district general hospital. PMID- 19311164 TI - Polydioxanone suture in the gastrointestinal tract. PMID- 19311166 TI - Recurrence rates following local anaesthetic day case inguinal hernia repair by junior surgeons in a district general hospital. PMID- 19311167 TI - Dorsal column stimulation (DCS) in chronic pain: report of 31 cases. PMID- 19311168 TI - Long term follow-up of patients with side to side choledochoduodenostomy and transduodenal sphincteroplasty: Assessor's Comments. PMID- 19311169 TI - Bupivacaine 'squirting'. PMID- 19311170 TI - Bupivacaine 'squirting'. PMID- 19311171 TI - A review of 155 extra-anatomic bypass grafts. PMID- 19311172 TI - A nutrition team is not essential for optimal parenteral nutrition therapy. PMID- 19311173 TI - A relationship between perioperative blood transfusion and recurrence of carcinoma of the sigmoid colon following potentially curative surgery. PMID- 19311174 TI - Preventing gastric regurgitation with a ballooned nasogastric tube. PMID- 19311175 TI - Surgical audit in a district general hospital: stimulus for improving patient care. PMID- 19311176 TI - The use of sterile adhesive tape in the closure of arthroscopic puncture wounds: a comparison with a single layer Nylon closure. PMID- 19311177 TI - The values of routine preoperative investigations. PMID- 19311178 TI - Editorial. PMID- 19311179 TI - Critical comment. PMID- 19311180 TI - British Association of Clinical Anatomists: Abstracts of papers presented at the Annual General Meeting on 5 January 1987. PMID- 19311181 TI - Bupivacaine squirting. PMID- 19311183 TI - The value and significance of the limited barium enema examination following restorative resection for carcinoma of the rectum. PMID- 19311182 TI - A relationship between perioperative blood transfusion and recurrence of carcinoma of the sigmoid colon following potentially curative surgery. PMID- 19311184 TI - A relationship between perioperative blood transfusion and recurrence of carcinoma of the sigmoid colon following potentially curative surgery. PMID- 19311185 TI - Surgical audit in a district general hospital. PMID- 19311186 TI - Fine needle aspiration cytology of breast masses. PMID- 19311187 TI - Duodenal ulcer perforation: the effect of H(2) antagonists? PMID- 19311188 TI - Colovesical fistula. PMID- 19311190 TI - Terence john millin-1903-80. PMID- 19311189 TI - Use of pedicled omentum in oesophagogastric anastomosis. PMID- 19311191 TI - The right approach to carcinoma of the cardia and lower oesophagus. PMID- 19311192 TI - Monitoring of renal allografts by Doppler ultrasound. PMID- 19311193 TI - To tunnel or not to tunnel catheters for parenteral nutrition. PMID- 19311194 TI - Removal of skin staples in an emergency. PMID- 19311196 TI - Colovesical fistula. PMID- 19311195 TI - The broken nose: does familiarity breed neglect? PMID- 19311197 TI - Is a drip necessary for a cholecystectomy? PMID- 19311198 TI - Complications of T-tube drainage of the common bile duct. PMID- 19311199 TI - A convenient aid for extension of the neck during thyroidectomy. PMID- 19311200 TI - A convenient aid for extension of the neck during thyroidectomy. PMID- 19311201 TI - The effect of dopamine on renal function during aortic cross clamping. PMID- 19311202 TI - The effect of dopamine on renal function during aortic cross clamping. PMID- 19311203 TI - Clinical acute cholecystitis and the Curtis-Fitz-Hugh syndrome. PMID- 19311204 TI - Philip Rowland Allison BSc ChM FRCS 1907-1974. PMID- 19311205 TI - A convenient aid for extension of the neck during thyroidectomy. PMID- 19311206 TI - The practical management of Fournier's gangrene. PMID- 19311207 TI - Changes in anal sphincter tone at induction of anaesthesia. PMID- 19311208 TI - Diverting staples (Removal of skin staples in an emergency). PMID- 19311209 TI - Bupivacaine squirting. PMID- 19311210 TI - Why not rub it better? PMID- 19311211 TI - The correlation between gross appearance of the appendix at appendicectomy and histological examination. PMID- 19311212 TI - Why on earth do surgeons need quality assurance? PMID- 19311214 TI - The correlation between gross appearance of the appendix at appendicectomy and histological examination. PMID- 19311213 TI - The correlation between gross appearance of the appendix at appendicectomy and histological examination. PMID- 19311215 TI - Bacterial colonisation of leg ulcers and its effect on the success rate of skin grafting. PMID- 19311216 TI - Limb salvage surgery in a district general hospital: factors affecting outcome. PMID- 19311217 TI - Bacterial colonisation of leg ulcers and its effect on the success rate of skin grafting. PMID- 19311218 TI - Clinical acute cholecystitis and the Curtis-Fitz-Hugh syndrome. PMID- 19311219 TI - Anaesthesia for major craniofacial surgery: a report of 23 cases in children under four years of age. PMID- 19311220 TI - Tuberculosis: a surgical viewpoint. PMID- 19311221 TI - Diagnosis of significant abdominal trauma after road traffic accidents. PMID- 19311222 TI - The effect of dopamine on renal function during aortic cross clamping. PMID- 19311223 TI - Transfer of anaesthetic records between hospitals. PMID- 19311224 TI - A convenient aid for the extension of the neck during thyroidectomy. PMID- 19311225 TI - The new operation codes: an improvement? PMID- 19311227 TI - Pancreatic sphincteroplasty: indications and outcome. PMID- 19311226 TI - The twist drain. PMID- 19311228 TI - Minitracheostomy-a report of a proposed further development. PMID- 19311229 TI - The twist drain. PMID- 19311230 TI - The broken nose: does familiarity breed neglect? PMID- 19311231 TI - Integrity of left-sided colorectal anastomoses. PMID- 19311233 TI - Minitracheostomy-a report of a proposed further development. PMID- 19311232 TI - Oesophageal resection in the elderly. PMID- 19311234 TI - Personality assessment techniques and aptitude testing as aids to the selection of surgical trainees. PMID- 19311235 TI - The cerebral complications of coronary artery bypass surgery. PMID- 19311236 TI - Unilateral lumbar facet joint hypertrophy causing nerve root irritation. PMID- 19311238 TI - An improved method for securing nasogastric tubes. PMID- 19311237 TI - An improved method for securing nasogastric tubes. PMID- 19311239 TI - Unilateral lumbar facet joint hypertrophy causing nerve root irritation. PMID- 19311240 TI - Early postoperative endoscopic sphincterotomy for retained biliary stones. PMID- 19311241 TI - Cardiac tamponade: a review of diagnosis and anaesthetic and surgical management illustrated by three case reports. PMID- 19311242 TI - Cardiac tamponade: a review of diagnosis and anaesthetic and surgical management illustrated by three case reports. PMID- 19311243 TI - Cardiac tamponade: a review of diagnosis and anaesthetic and surgical management illustrated by three case reports. PMID- 19311244 TI - Editorial. PMID- 19311245 TI - Intra-operative air testing: an audit on rectal anastomosis. PMID- 19311247 TI - 168 double J (pigtail) ureteric catheter insertions: a retrospective review. PMID- 19311246 TI - An improved method for securing nasogastric tubes. PMID- 19311248 TI - Protection for the left index finger whilst operating on HIV positive patients. PMID- 19311249 TI - Surgery for gastro-oesophageal reflux: the Angelchik prosthesis compared to the floppy Nissen fundoplication. Two-year follow-up study and a five-year evaluation of the Angelchik prosthesis: Assessor's comments. PMID- 19311250 TI - An operation for rectal stricture following Ripstein rectopexy. PMID- 19311251 TI - Do children need routine preoperative blood tests and blood cross matching in orthopaedic practice? PMID- 19311252 TI - Subclavian vein catheterisation for parenteral nutrition. PMID- 19311253 TI - Intra-operative air testing: an audit on rectal anastomosis. PMID- 19311255 TI - The functional result of pelvic ileal reservoir in 10 patients with familial adenomatous polyposis. PMID- 19311254 TI - Pancreatic sphincteroplasty: indications and outcome. PMID- 19311256 TI - Cholecystectomy and the development of colorectal neoplasia: a prospective study. PMID- 19311257 TI - Cholecystectomy and the development of colorectal neoplasia: a prospective study. PMID- 19311258 TI - Acute traumatic haemarthrosis of the knee: expectant treatment or arthroscopy? PMID- 19311259 TI - Clean up varicose vein surgery-use a tourniquet. PMID- 19311261 TI - Integrity of left-sided colorectal anastomoses. PMID- 19311260 TI - Clean up varicose vein surgery-use a tourniquet. PMID- 19311262 TI - To stitch or not to stitch the fat. PMID- 19311263 TI - To stitch or not to stitch the fat. PMID- 19311264 TI - Surgery in a geriatric population. PMID- 19311265 TI - Cardiac complications of total parenteral nutrition: the role of two-dimensional echocardiography in diagnosis. PMID- 19311266 TI - The natural history of peptic oesophageal strictures treated by dilatation and antireflux therapy alone: Assessor's comments. PMID- 19311267 TI - Massive colonic haemorrhage-the case for right hemicolectomy. PMID- 19311268 TI - To stitch or not to stitch the fat? PMID- 19311269 TI - Editorial. PMID- 19311270 TI - 101 Oesophageal cancers: a surgeon uses radiotherapy: Assessor's comment. PMID- 19311271 TI - Posterior tibial tendon rupture-a brief report. PMID- 19311273 TI - A new technique of caecostomy using endotracheal tubes. PMID- 19311272 TI - Postoperative pain relief; a new approach: narcotics compared with non-steroidal anti-inflammatory drugs. PMID- 19311274 TI - Plasma lignocaine levels during transurethral prostatectomy. PMID- 19311275 TI - An operation for rectal strictures following Ripstein rectopexy. PMID- 19311276 TI - Varicose vein surgery using a pneumatic tourniquet: reduced blood loss and improved cosmesis: Assessor's Comment. PMID- 19311277 TI - Catheterisation for nephrectomy? PMID- 19311278 TI - Early postoperative endoscopic sphincterotomy for retained common duct stones. PMID- 19311279 TI - Read this! PMID- 19311280 TI - Junior medical staff and the assessment of trauma: Assessor's comment. PMID- 19311281 TI - Amputation for antipersonnel mine injuries of the leg: preservation of the tibial stump using a medial gastrocnemius myoplasty. PMID- 19311282 TI - Postoperative pain relief; a new approach: narcotics compared with non-steroidal anti-inflammatory drugs. PMID- 19311283 TI - Postoperative pain relief; a new approach: narcotics compared with non-steroidal anti-inflammatory drugs. PMID- 19311285 TI - Drainage or suture of the cavity after breast biopsy. PMID- 19311284 TI - The application of isotope limb blood flow measurement to diagnostic problems in vascular surgery. PMID- 19311286 TI - A comparison of postoperative pain relief techniques in orchidopexy. PMID- 19311287 TI - The perimedian incision. PMID- 19311289 TI - The perimedian incision. PMID- 19311288 TI - Intraoperative assessment of lymph node involvement in gastric carcinoma. PMID- 19311290 TI - Optimal operative treatment in acute septic complications of diverticular disease. PMID- 19311291 TI - The perimedian incision. PMID- 19311292 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311293 TI - Mastectomy and breast reconstruction preserving the nipple. PMID- 19311294 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311296 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311295 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311297 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311298 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311299 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311300 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311301 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311302 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311303 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311304 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311305 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311306 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311307 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311308 TI - Axillary block anaesthesia in acute and elective hand surgery: a report on 300 procedures. PMID- 19311309 TI - The incidence of congenitally absent foot pulses. PMID- 19311310 TI - A simple technique for successful primary closure after excision of pilonidal sinus disease: Assessor's comment. PMID- 19311311 TI - Local analgesia for infant pyloromyotomy. Does wound infiltration with bupivacaine affect postoperative behaviour? Assessor's comment. PMID- 19311312 TI - Thoracoabdominal total gastrectomy in the management of adenocarcinoma of the cardia. Is it worth it? Assessor's comment. PMID- 19311313 TI - Aortocaval fistulas and the use of transvenous balloon tamponade: Assessor's comment. PMID- 19311314 TI - Lower oesophageal contractility monitoring during anaesthesia for cardiac surgery: preliminary observations. PMID- 19311315 TI - Drainage or suture of the cavity after breast biopsy. PMID- 19311316 TI - Blood transfusion and surgery: the effect on growth of a syngeneic sarcoma. PMID- 19311317 TI - Junior medical staff and the assessment of trauma. PMID- 19311318 TI - Intra-articular and subcutaneous prilocaine with adrenaline for pain relief in day case arthroscopy of the knee joint: Assessor's comment. PMID- 19311319 TI - Antiseptic toxicity to breast carcinoma in tissue culture: an adjuvant to conservation therapy? Assessor's comment. PMID- 19311320 TI - The presentation of malrotation of the intestine in adults. PMID- 19311321 TI - The use of computed tomography in the evaluation of large multinodular goitres: Assessor's comment. PMID- 19311322 TI - Transcholecystic operative cholangiography: an alternative technique: Assessor's comment. PMID- 19311323 TI - Blood transfusion in total hip replacement: Is it always necessary? Assessor's comment. PMID- 19311324 TI - Percutaneous transluminal angioplasty of the subclavian artery: Assessor's comment. PMID- 19311325 TI - Deaths following trauma: an audit of performance: Comment. PMID- 19311326 TI - Catheterisation for nephrectomy. PMID- 19311328 TI - McBurney's point - fact or fiction? PMID- 19311329 TI - McBurney's point - fact or fiction? PMID- 19311331 TI - Chest wall reconstruction after resection of recurrent breast tumours: Assessor's comment. PMID- 19311330 TI - McBurney's point - fact or fiction? PMID- 19311332 TI - Effect of blood transfusion on survival after radiotherapy as treatment for carcinoma of the prostate: Assessor's comment. PMID- 19311333 TI - Suturing of digital lacerations: digital block or local infiltration? PMID- 19311334 TI - Factors influencing peritoneal catheter survival in continuous ambulatory peritoneal dialysis. PMID- 19311336 TI - Outpatient carpal tunnel decompression without tourniquet: a simple local anaesthetic technique. PMID- 19311335 TI - Outpatient carpal tunnel decompression without tourniquet: a simple local anaesthetic technique. PMID- 19311337 TI - Arthroscopic meniscal shavers: a potential hazard of sepsis: Assessor's comment. PMID- 19311339 TI - Selective arterial embolisation for hepatic trauma: Assessor's comment. PMID- 19311338 TI - Early post-splenectomy sepsis after missile injury in adults: Assessor's comment. PMID- 19311340 TI - Uncommon complications of laparoscopic sterilisation. PMID- 19311341 TI - Junior medical staff and the assessment of trauma. PMID- 19311342 TI - Deaths following trauma: an audit of performance. PMID- 19311343 TI - A comparative study between Michel and Proximate clips for the closure of neck incisions: Assessor's comment. PMID- 19311344 TI - Audit of attitudes to and use of postoperative thromboembolic prophylaxis in a regional health authority: Assessor's comment. PMID- 19311345 TI - The varicose vein waiting list: results of a validation exercise: Assessor's comment. PMID- 19311346 TI - The incidence and value of rectal examination in children with suspected appendicitis: Assessor's comment. PMID- 19311348 TI - Cutaneous malignant melanoma: audit of the diagnostic process: Assessor's comment. PMID- 19311347 TI - Incidence and prevention of conjunctival contamination with blood during hazardous surgical procedures: Assessor's comment. PMID- 19311349 TI - The role of reconstructive surgery in the management of war wounds. PMID- 19311350 TI - Deaths following trauma: an audit of performance. PMID- 19311351 TI - Duration of intravenous fluid replacement after abdominal surgery: a prospective randomised study. PMID- 19311353 TI - An audit of surgical emergencies in the very old: Assessor's comment. PMID- 19311354 TI - Control of presacral bleeding in rectal surgery: Assessor's comment. PMID- 19311355 TI - Nasogastric suction after elective abdominal surgery: a randomised study: Assessor's comment. PMID- 19311356 TI - Which general surgical operations must be done at night? Assessor's Comment. PMID- 19311357 TI - Mobile versus fixed site lithotripsy: Assessor's Comment. PMID- 19311358 TI - Orthopaedic trauma in men: the relative risk among drinkers and the prevalence of problem drinking in male orthopaedic admissions: Assessor's Comment. PMID- 19311359 TI - A controlled trial of short-term versus standard axillary drainage after axillary clearance and iridium implant treatment of early breast cancer: Assessor's Comment. PMID- 19311360 TI - Amputations in diabetics. PMID- 19311361 TI - Malignant neck lumps: a measured approach. PMID- 19311362 TI - Editorial. PMID- 19311364 TI - Non-steroidal anti-inflammatory drugs and stercoral perforation of the colon: Assessor's comment. PMID- 19311365 TI - An objective long-term evaluation of the Angelchik antireflux prosthesis: Assessor's comment. PMID- 19311366 TI - Breast cancer data collection for surgical audit: Assessor's comment. PMID- 19311367 TI - Sedation for day-case urology: an assessment of patient recovery profiles after midazolam and flumazenil: Assessor's comment. PMID- 19311369 TI - Use of operating theatres: the effects of case-mix and training in general surgery: Assessors' comments. PMID- 19311368 TI - Postoperative collection and reinfusion of autologous blood in total knee arthroplasty: Assessor's comment. PMID- 19311370 TI - Use of operating theatres: the effects of case-mix and training in general surgery: Assessors' comments. PMID- 19311372 TI - Historical aspects of amputation. PMID- 19311371 TI - Surgical options for left-sided large bowel emergencies: Assessor's comment. PMID- 19311373 TI - Ultrasound guided subclavian vein catheterisation. PMID- 19311374 TI - A comparative study between Michel and Proximate clips for closure of neck incisions. PMID- 19311375 TI - The incidence and value of rectal examination in children with suspected appendicitis. PMID- 19311376 TI - CA15-3: a reliable indicator of metastatic bone disease in breast cancer patients: Assessor's comment. PMID- 19311377 TI - Endoprosthetic replacement for bony metastases: Assessor's comment. PMID- 19311378 TI - Paraoesophageal hiatus hernia: surgery for all ages: Assessor's comment. PMID- 19311379 TI - Forty-four years on. PMID- 19311380 TI - Simulated laparoscopic cholecystectomy: Assessor's comment. PMID- 19311381 TI - A comparative study between Michel and Proximate clips for the closure of neck incisions. PMID- 19311382 TI - Prevention of occupational transmission of HIV in the ENT clinic: Assessor's comment. PMID- 19311383 TI - Surgical debate-Intra-arterial thrombolysis should be the initial treatment of the acutely ischaemic lower limb: Assessor's comment. PMID- 19311384 TI - Buerger's disease and cigarette smoking in Bangladesh: Assessor's comment. PMID- 19311385 TI - Chronic pancreatitis with biliary obstruction: Assessors' comments. PMID- 19311387 TI - Radical axillary dissection in the staging and treatment of breast cancer: Assessor's comment. PMID- 19311386 TI - Chronic pancreatitis with biliary obstruction: Assessors' comments. PMID- 19311388 TI - Day-case paediatric surgery: the only choice: Assessor's comment. PMID- 19311389 TI - Serum gastrin concentrations in colorectal cancer patients: Assessor's comment. PMID- 19311390 TI - Angiomyolipoma of the kidney: a clinical enigma in diagnosis and management: Assessor's comment. PMID- 19311391 TI - The challenge of evaluating surgical procedures: Assessor's comment. PMID- 19311392 TI - The ORMA retractor holder: Assessor's comment. PMID- 19311393 TI - An insight into Dupuytren's contracture: Assessor's comment. PMID- 19311394 TI - The prognostic value of nucleolar organiser regions in colorectal cancer: a 5 year follow-up study: Assessor's comment. PMID- 19311396 TI - Non-specific abdominal pain: the resource implications: Assessor's comment. PMID- 19311395 TI - Does surgical experience influence mastectomy complications: Assessor's comment. PMID- 19311397 TI - Carcinoma of the major salivary glands: Assessor's comment. PMID- 19311398 TI - Use of disposable skin staplers for bowel anastomosis to reduce laparotomy time in war: Assessor's comment. PMID- 19311399 TI - Audit of 200 consecutive aortic aneurysm repairs carried out by a single surgeon in a district hospital: results of surgery and factors affecting outcome: Assessor's comment. PMID- 19311400 TI - Care of road traffic accident victims in a district general hospital: Assessor's comment. PMID- 19311401 TI - Branchial cysts: congenital or acquired? PMID- 19311402 TI - Prevention of occupational transmission of HIV in the ENT clinic. PMID- 19311403 TI - Paraoesophageal hiatus hernia: surgery for all ages. PMID- 19311404 TI - Endoscopic transanal resection of large villous tumours of the rectum. PMID- 19311405 TI - Assessment of the use of disposable skin staplers to reduce laparotomy time in penetrating ballistic injury of the abdomen. PMID- 19311406 TI - The diagnosis of soft tissue tumours: Invited comment. PMID- 19311408 TI - Measuring outcome of total knee replacement using quality of life indices: Invited comment. PMID- 19311407 TI - A simple scoring system to reduce the negative appendicectomy rate rate: Invited comment. PMID- 19311409 TI - Changing patterns of colorectal cancer in a regional teaching hospital: Invited comment. PMID- 19311411 TI - The ORMA retractor holder. PMID- 19311412 TI - Simulated laparoscopic cholecystectomy. PMID- 19311413 TI - Non-tumour morbidity and mortality after modified radical mastectomy: Invited comment. PMID- 19311414 TI - Limb exsanguination. II. The leg: effect of angle of elevation: Invited comment. PMID- 19311415 TI - Prevention of bacterial infection and sepsis in acute severe pancreatitis: Invited comment. PMID- 19311416 TI - Plasma amylase estimation in recurrent abdominal pain in children: Invited comment. PMID- 19311417 TI - Delayed intracerebral haematomas in moderate to severe head injuries in young adults: Invited comment. PMID- 19311418 TI - Intestinal ischaemia in the unconscious intensive care unit patient: Invited comment. PMID- 19311419 TI - Blood loss and transfusion requirements in total joint arthroplasty: Invited comment. PMID- 19311420 TI - Acute colonic pseudo-obstruction: a pharmacological approach: Invited comment. PMID- 19311421 TI - Editorial-Palpation of peripheral pulses: a difficult art. PMID- 19311423 TI - Costs can and must be a component of surgical audit: Invited comment. PMID- 19311424 TI - Activity analysis: measurement of the effectiveness of surgical training and operative technique: Invited comment. PMID- 19311425 TI - A 5-year review of carotid endarterectomy in a vascular unit using a computerised audit system: Invited comment. PMID- 19311426 TI - Will screening for breast cancer reduce mortality? Evidence from the first year of screening in Avon: Invited comment. PMID- 19311427 TI - Infantile hypertrophic pyloric stenosis: where should it be treated?: Invited comment. PMID- 19311429 TI - Laparoscopic operations. PMID- 19311428 TI - Limb exsanguination. PMID- 19311430 TI - Cutaneous perianal recurrence of cancer after anterior resection using the EEA stapling device. PMID- 19311431 TI - Phytobezoar: an uncommon cause of small bowel obstruction. PMID- 19311433 TI - A prospective comparison of laparoscopic versus open cholecystectomy. PMID- 19311432 TI - Fractures of the sternal body. PMID- 19311434 TI - A prospective comparison of laparoscopic versus open cholecystectomy. PMID- 19311436 TI - Comparison of 'intraoperative' parathormone measurement with frozen section during parathyroid surgery. PMID- 19311435 TI - Editorial-The Annals: the editorial process revealed. PMID- 19311437 TI - Blood transfusion does not have an adverse effect on survival after operation for colorectal cancer: Invited comment. PMID- 19311438 TI - Provision and acceptability of day case breast biopsy: an audit of current practice: Invited comment. PMID- 19311439 TI - Infantile hypertrophic pyloric stenosis: where should it be treated? PMID- 19311440 TI - Colostomy is no longer appropriate in the management of uncomplicated large bowel obstruction: true or false? PMID- 19311441 TI - Surgical discharge summaries: improving the record. PMID- 19311442 TI - The use of Coca-Cola in the management of bolus obstruction in benign oesophageal stricture. PMID- 19311443 TI - Surgical audit without autopsy: tales of the unexpected. PMID- 19311445 TI - Submandibular gland surgery: an audit of clinical findings, pathology and postoperative morbidity. PMID- 19311444 TI - Submandibular gland surgery: an audit of clinical findings, pathology and postoperative morbidity. PMID- 19311446 TI - Non-woven, disposable theatre gowns for 'high-risk' surgery. PMID- 19311447 TI - Provision and acceptability of day case breast biopsy: an audit of current practice. PMID- 19311448 TI - Editorial note: Intravenous sedation in accident and emergency departments: a nationwide survey. PMID- 19311449 TI - A trauma centre in the UK. PMID- 19311450 TI - Trauma audit: clinical judgement or statistical analysis? PMID- 19311451 TI - Post-cholecystectomy symptoms after laparoscopic cholecystectomy. PMID- 19311453 TI - Editorial comment. PMID- 19311452 TI - Reply by the chairman of the working party. PMID- 19311455 TI - Loose marker tapes. PMID- 19311454 TI - How long do patients convalesce after inguinal herniorrhaphy? Current principles and practice. PMID- 19311456 TI - Shaving patients before operation: a dangerous myth? PMID- 19311458 TI - Hand infections secondary to fish bone injuries. PMID- 19311457 TI - Operative lumbar sympathectomy for severe lower limb ischaemia: still a valuable treatment option. PMID- 19311459 TI - Single axillary crease incision for wide local excision and axillary clearance in breast cancer. PMID- 19311460 TI - The Read clinical classification and its use in plastic surgery. PMID- 19311461 TI - An orthopaedic theatre timings survey. PMID- 19311462 TI - Emergency workload in otolaryngology. PMID- 19311464 TI - Pelvic abscess following endoscopic appendicectomy. PMID- 19311463 TI - Experience with a knot-free absorbable subcuticular suture. PMID- 19311465 TI - Sir Henry souttar. PMID- 19311466 TI - Experience of the knot-free absorbable subcutaneous suture. PMID- 19311467 TI - The surgery of mitral stenosis 1898-1948: why did it take 50 years to establish mitral valvotomy? PMID- 19311468 TI - Long posteromedial myocutaneous flap below-knee amputation. PMID- 19311469 TI - Use of split-skin grafting in the treatment of chronic leg ulcers. PMID- 19311470 TI - Acute scybalous colonic obstruction and perforation. PMID- 19311472 TI - Editorial-Times of change: the College the Annals and the Bulletin. PMID- 19311471 TI - Crochet hooks in varicose vein surgery. PMID- 19311473 TI - What is clinical audit?: Comment. PMID- 19311475 TI - Antipersonnel landmines. PMID- 19311474 TI - Morbidity of varicose vein surgery: auditing the benefit of changing clinical practice. PMID- 19311476 TI - An effective shield for free: pulsed lavage in total knee replacement. PMID- 19311477 TI - Transecting the rectum: the 'clean cut' approach. PMID- 19311478 TI - Audit of pain after nasal surgery. PMID- 19311479 TI - Hepatitis B vaccine uptake among surgeons at a London teaching hospital: how well are we doing? PMID- 19311480 TI - What is clinical audit? PMID- 19311481 TI - A novel method for the treatment of localised intrathoracic anastomotic leakage. PMID- 19311482 TI - Lower gastrointestinal bleeding during anticoagulant therapy: a life-saving complication? PMID- 19311483 TI - Role of fine needle aspiration cytology in the management of the discrete parotid lump. PMID- 19311484 TI - Editorial-The next 50 years. PMID- 19311486 TI - The Didcott dilator. PMID- 19311485 TI - ERRATUM: Contents. PMID- 19311488 TI - Intercollegiate specialty examinations - 2000. PMID- 19311487 TI - The case against a national screening programme for aortic aneurysm: Overview. PMID- 19311489 TI - Editorial. PMID- 19311490 TI - The royal society of medicine: section of surgery, medical insurance agency prize 1999. PMID- 19311491 TI - RSCE response. PMID- 19311493 TI - Editorial. PMID- 19311492 TI - Council election. PMID- 19311494 TI - The need of hospital provision for chronic patients. PMID- 19311495 TI - The hospital situation in Greater New York. PMID- 19311496 TI - Report. PMID- 19311498 TI - Morris Harold Frantz. PMID- 19311497 TI - John W. Small. PMID- 19311499 TI - William McKay. PMID- 19311500 TI - William E. Porter. PMID- 19311501 TI - Walter M. Silleck. PMID- 19311502 TI - John James Cotter. PMID- 19311504 TI - Samuel Bard and the King's College School. PMID- 19311503 TI - Sketch of John Wakefield Francis. PMID- 19311505 TI - Historical aspects of diabetes and insulin. PMID- 19311506 TI - Report on the indiscriminate use of X-rays in the treatment of hypertrichosis. PMID- 19311507 TI - Opportunities for graduate medical study in New York. PMID- 19311508 TI - Serum reactions in Syphilis. PMID- 19311509 TI - The nursing situation. PMID- 19311510 TI - Hosack Hall and the Hosack hospital bed for sick and needy physicians. PMID- 19311511 TI - David Hosack. PMID- 19311512 TI - The Application of Newer Methods in Blood-Chemistry to Clinical Medicine. PMID- 19311513 TI - The Use of Iodin in the Treatment of Diseases of the Thyroid Gland. PMID- 19311514 TI - On the Mechanism of Insulin Action. PMID- 19311515 TI - Valentine Mott. PMID- 19311516 TI - Functions of the Academy. PMID- 19311517 TI - Functions of the Academy of Medicine of Paris. PMID- 19311518 TI - Convalescent Care. PMID- 19311519 TI - Summary report on institutional convalescence. PMID- 19311521 TI - The Finances of the Academy. PMID- 19311520 TI - Alonzo Clark. PMID- 19311522 TI - The Charcot Centenary. PMID- 19311523 TI - On the understanding and practical management of nervous patients, particularly of the nervous woman. PMID- 19311524 TI - 1. Address of the President, Dr. Samuel A. Brown. PMID- 19311526 TI - Library of the Medical Society of the County of Kings. PMID- 19311525 TI - 2. Address of Dr. George David Stewart. PMID- 19311527 TI - Huxley After Thirty Years. PMID- 19311528 TI - The destruction of the quarantine station on Staten Island in 1858. PMID- 19311529 TI - Proposed changes in the Constitution and By-laws of The New York Academy of Medicine. PMID- 19311530 TI - Consolidation of municipal hospitals. PMID- 19311531 TI - The Charcot Exhibit. PMID- 19311532 TI - The Academy of Medicine as a Prime Mover in Quarantine Legislation. PMID- 19311533 TI - Internships in the American Hospital in Paris. PMID- 19311534 TI - Synopses on orthopedic surgery and neurology and psychiatry. PMID- 19311535 TI - The history of cancer. PMID- 19311536 TI - Some considerations of the mode of infection and extension of erysipelas. PMID- 19311537 TI - Pruritus, causes, effects and treatment. PMID- 19311538 TI - Section of Otology, March 12th: X-Ray Examination as an Aid to Mastoid Surgery. PMID- 19311540 TI - The needs of the Library. PMID- 19311539 TI - Directory of convalescent homes. PMID- 19311542 TI - Psychiatric service at Bellevue Hospital. PMID- 19311541 TI - Surgical criteria for cholecystectomy. PMID- 19311543 TI - The reticulo-endothelial system. PMID- 19311545 TI - Testimonial dinner to Dr. Dana. PMID- 19311544 TI - The Temples of Aesculapius. PMID- 19311547 TI - The object, future development and financial needs of the Academy. PMID- 19311546 TI - Laennec. PMID- 19311548 TI - Carbon monoxide poisoning. PMID- 19311550 TI - Summary report on cardiac children. PMID- 19311549 TI - Bacon's relation to medicine. PMID- 19311552 TI - Observations during the period of undernutrition in Germany. PMID- 19311551 TI - Series and families of diseases. PMID- 19311554 TI - The inscriptions on the new building. PMID- 19311553 TI - Maude Abbott's Osler Memorials. PMID- 19311555 TI - SUPPLEMENT. PMID- 19311556 TI - The Gesolei at Dusseldorf. PMID- 19311557 TI - Medical visitors to New York. PMID- 19311559 TI - Amendments to the Constitution and By-Laws. PMID- 19311558 TI - Opening of the new building. PMID- 19311561 TI - The history of heating, ventilation and lighting. PMID- 19311560 TI - The Wesley M. Carpenter Lecture: Ionization and chemical reactions. PMID- 19311562 TI - Addresses delivered at the dinner in honor of the newly elected Honorary Fellows. PMID- 19311564 TI - Addresses. PMID- 19311563 TI - Dedicatory prayer. PMID- 19311565 TI - Bacteriophagy and the theory of ultrascopic viruses. PMID- 19311567 TI - Report on ambulances. PMID- 19311566 TI - Report on the administration of the City Health Department. PMID- 19311569 TI - Catalogue of an exhibition of works in the plastic and graphic arts by American physicians. PMID- 19311568 TI - New Academy committees. PMID- 19311570 TI - The development of military medicine. PMID- 19311571 TI - The Philosophy of History. PMID- 19311572 TI - Address. PMID- 19311573 TI - Constitution and By-laws. PMID- 19311574 TI - Opinion on the Internal Revenue Act. PMID- 19311575 TI - Hosack Bed for Sick and Needy Physicians. PMID- 19311576 TI - Constitution and Characterology. PMID- 19311577 TI - Dinner to Sir George Newman. PMID- 19311578 TI - Recommendations Relative to the Contagious Disease Hospitals of the Department of Health of the City of New York. PMID- 19311579 TI - Protein Stimulation. PMID- 19311580 TI - Resolution in regard to Volstead Act. PMID- 19311581 TI - Opportunities for Graduate Medical Study in Hospitals of New York City. PMID- 19311582 TI - Early Medieval Anatomy. PMID- 19311583 TI - The Future Food Supply of the United States. PMID- 19311584 TI - Cardiac Asthma. PMID- 19311585 TI - The Myasthenic Syndrome. PMID- 19311586 TI - How Young Doctors Should Behave. PMID- 19311587 TI - Incidents in the Recent History of The New York Academy of Medicine. PMID- 19311588 TI - Colonial Medicine, or the Doctor of Our Forefathers. PMID- 19311589 TI - Resolution of the Council: Dr. Herbert S. Carter. PMID- 19311590 TI - Report on the Chief Medical Examiner's Office. PMID- 19311591 TI - Action of the Trustees in regard to a bequest. PMID- 19311592 TI - Impressions of medical teaching and hospital work. PMID- 19311593 TI - Convalescent care. PMID- 19311594 TI - Recent Knowledge of Epidemic Diseases. PMID- 19311595 TI - Body and Mind. PMID- 19311597 TI - Summary of Activities for Month of January. PMID- 19311598 TI - II. Clinical Results Following Malarial Therapy in General Paresis. PMID- 19311596 TI - A Bit of Financial History of the Academy. PMID- 19311599 TI - III. Experiences With Malarial Inoculation in Syphilis. PMID- 19311600 TI - On the Significance of Bacterial Allergy in Infectious Diseases. PMID- 19311602 TI - Testimonial dinner to Dr. Norris. PMID- 19311601 TI - Annual Graduate Fortnight. PMID- 19311603 TI - Summary of activities for the month of February. PMID- 19311604 TI - A Neglected Medical Scholar. PMID- 19311605 TI - Animal Reservoirs of Human Disease with Special Reference to Microbic Variability. PMID- 19311607 TI - Organizing Convalescent Care. PMID- 19311606 TI - Napoleon-His Last Illness and Postmortem. PMID- 19311608 TI - Available Sources and Future Prospects of Medical Biography. PMID- 19311609 TI - Presentation of portrait of Dr. William H. Park. PMID- 19311610 TI - Activities for the months of March and April. PMID- 19311611 TI - Practical Lecture Series 1928-29. PMID- 19311613 TI - Medical Information Bureau. PMID- 19311612 TI - Streeter collection acquired. PMID- 19311614 TI - Trustees and Council. PMID- 19311615 TI - The New York Academy of Medicine Medal. PMID- 19311616 TI - Exhibition of works of art by medical men. PMID- 19311618 TI - Annual Graduate Fortnight. PMID- 19311617 TI - Maternal Mortality. PMID- 19311620 TI - The First Graduate Fortnight. PMID- 19311619 TI - Jefferson Medical College and Hospital. PMID- 19311621 TI - The Future of Surgery, May 3, 1928. PMID- 19311622 TI - Editorial: Naunyn's Autobiography. PMID- 19311623 TI - The Graduate Fortnight. PMID- 19311624 TI - The Border Line of Ethics. PMID- 19311625 TI - The Pathology of the Aging Process. PMID- 19311626 TI - Medical Proverbs, Aphorisms and Epigrams. PMID- 19311627 TI - Introductory Remarks. PMID- 19311628 TI - The Pains, Penalties and Prohibitions of Old Age. Can They Be Prevented? PMID- 19311629 TI - Old Age and What It Means to the Community. PMID- 19311630 TI - Dietetics in Old Age. PMID- 19311632 TI - Alcohol and Old Age. PMID- 19311631 TI - Pharmacology in Old Age. PMID- 19311633 TI - The Aging of the Human Brain. PMID- 19311635 TI - The Diagnosis and Treatment of Preparalytic Poliomyelitis. PMID- 19311634 TI - Extract from a Report of the Sub-Committee in Charge of the Graduate Fortnight. PMID- 19311636 TI - The International Hospital Congress. PMID- 19311637 TI - Annual Report of the Hospital Information and Service Bureau. PMID- 19311638 TI - Clinical Aspect and Management of Old Age from the Practitioner's Point of View. PMID- 19311639 TI - Climate and the Aged. PMID- 19311640 TI - Psychoses in Old Age. PMID- 19311642 TI - Apoplexy. PMID- 19311641 TI - Present Status of the Problem of the So-Called Rejuvenation. PMID- 19311643 TI - Excerpt from Council Minutes, November 28, 1928. PMID- 19311645 TI - Relation of Endocrine Disorders to Senescence. PMID- 19311646 TI - Arterial Diseases of the Brain and Cord. PMID- 19311647 TI - Menopausal and Post-Menopausal Conditions in Women. PMID- 19311648 TI - Urology, its Contacts with General Medicine. PMID- 19311649 TI - Arthritis and Old Age. PMID- 19311650 TI - Traumatic Surgery and the Problems of Age. PMID- 19311651 TI - Diseases of the Eye in Old Age. PMID- 19311652 TI - The Evil Spoken of Physicians and the Answer Thereto. PMID- 19311653 TI - Address of the Retiring President: The Academy as an Educational Institution. PMID- 19311654 TI - Address of the Incoming President: The Academy, the Profession and the Public. PMID- 19311655 TI - A Study of the Streptococcus in the Etiology of Arthritis. PMID- 19311656 TI - A Clinical Index of Malignancy for Carcinoma of the Breast. PMID- 19311657 TI - Public Health Relations Committee: Report on Open Hospitals in New York City. PMID- 19311659 TI - The Skin as a Shock Tissue. PMID- 19311658 TI - Committee on Medical Education: Second Annual Graduate Fortnight. PMID- 19311660 TI - Allergy in Skin Diseases. PMID- 19311661 TI - Salpingitis. PMID- 19311662 TI - Lung Abscess-Some Aspects of Etiology and Medical Treatment. PMID- 19311664 TI - Catalogue of Medical Manuscripts and Incunabula. PMID- 19311663 TI - Excerpts from the Minutes of the Council. PMID- 19311666 TI - Medicine as an Agency in the Advancement of Science, Art and Civilization. PMID- 19311665 TI - Catalogue of Art Exhibit. PMID- 19311667 TI - The Serum Treatment and its Evaluation in Lobar Pneumonia. PMID- 19311668 TI - To the Great Clinician Friedrich von Muller on His Seventieth Birthday. PMID- 19311669 TI - Osleriana. PMID- 19311670 TI - Supervisory Powers of the State Board of Charities. PMID- 19311671 TI - Resolution in Honor of Dr. Dana. PMID- 19311672 TI - Resolution of the Council on the Death of Dr. Widal. PMID- 19311674 TI - Control of Conception, Present and Future. PMID- 19311673 TI - A Medical Tour in the West. PMID- 19311676 TI - Modern Methods of Resuscitation in New York City. PMID- 19311675 TI - The Centenary of Johann Nepomuk Czermak. PMID- 19311677 TI - Committee on Professional Standards. PMID- 19311678 TI - Protection of Patients' Confidential Records. PMID- 19311679 TI - Ramon y Cajal. PMID- 19311680 TI - The Medicine of the American Indian. PMID- 19311681 TI - Segmental Hyperalgesia and Segmental Increased Muscle Tone in Diseases of the Lungs and Heart. PMID- 19311683 TI - Action of the Council. PMID- 19311682 TI - Unveiling of Memorial Tablet to Dr. Holt. PMID- 19311684 TI - What Medicine Can Do for Law. PMID- 19311685 TI - The Role of the Medical Expert in Criminal Trials. PMID- 19311686 TI - Social Danger of the Borderline Mental Case. PMID- 19311688 TI - Second Graduate Fortnight. PMID- 19311687 TI - Medico-Legal Problems. PMID- 19311689 TI - Classification of the Medical Positions under the Municipal Civil Service of the City of New York, as recommended by The Public Health Relations Committee of The New York Academy of Medicine. PMID- 19311690 TI - Developmental Possibilities in Medical History as a Branch of the Medical Curriculum. PMID- 19311691 TI - The Common Affections of the Colon, their Origin and their Management. PMID- 19311693 TI - Second Graduate Fortnight. PMID- 19311692 TI - History of Post-Graduate Medicine in New York City. PMID- 19311694 TI - Public Welfare in Vienna. PMID- 19311695 TI - The Genesis of the Medical Department of the United States Army. PMID- 19311696 TI - Allusions to Medicine in Classical Literature. PMID- 19311697 TI - Second Annual Graduate Fortnight. PMID- 19311698 TI - MEMORIAL ADDRESSES: Hideyo Noguchi. PMID- 19311699 TI - The History of Drainage, Irrigation, Sewage-Disposal and Water-Supply. PMID- 19311700 TI - Allergy as Encountered by the General Practitioner. PMID- 19311701 TI - Some Needs in Medical Bibliography. PMID- 19311702 TI - Resolutions of the Council Adopted October 30, 1929. PMID- 19311704 TI - Address of Welcome. PMID- 19311703 TI - The Importance of the Emotions in the Etiology and Prognosis of Disease. PMID- 19311705 TI - The Graduate Fortnight. PMID- 19311706 TI - The Origin and Growth of the Mental Hygiene Movement. PMID- 19311707 TI - The Involuntary Nervous System. PMID- 19311708 TI - Hysteria as a Practical Problem. PMID- 19311709 TI - Neuroses Following Accident. PMID- 19311710 TI - The Surgical Indications for Sympathetic Ganglionectomy and Trunk Resection in the Treatment of Diseases Resulting from Vasomotor Spasm of Peripheral Arteries. PMID- 19311711 TI - The Endocrines and the Vegetative Nervous System. PMID- 19311712 TI - Legal Status of Hospital Records. PMID- 19311713 TI - Medicine in India. PMID- 19311714 TI - Award of the Academy Medal. PMID- 19311715 TI - Headache and Migraine. PMID- 19311716 TI - The Cardiac Neuroses. PMID- 19311717 TI - The Psychoneuroses Affecting the Gastro-Intestinal Tract. PMID- 19311719 TI - Council Resolution. PMID- 19311718 TI - Postoperative Emotional Disorders: Their Prevention and Management. PMID- 19311720 TI - The Spas Committee-Summary of the Report and Recommendations. PMID- 19311721 TI - Catalogue of Art Exhibit. PMID- 19311722 TI - Neurocirculatory Asthenia. PMID- 19311723 TI - A Medical Tour in Europe. PMID- 19311724 TI - Opportunities Which New York City Offers for Post-Graduate Medical Study. PMID- 19311726 TI - In Memory of Manson. PMID- 19311725 TI - The Edward N. Gibbs Memorial Prize Fund. PMID- 19311727 TI - 1. Psychotherapy. PMID- 19311729 TI - 3. General Survey of Visceral Neuroses. PMID- 19311728 TI - 2. The Psychoneuroses. PMID- 19311731 TI - Outline of Preventive Medicine. PMID- 19311730 TI - 4. Insomnia and Disturbances of Sleep. PMID- 19311732 TI - Recent Publications of the Committee on Medical Education. PMID- 19311734 TI - American German Medical Exchange Opportunities. PMID- 19311733 TI - Progress in the Cataloguing of Medical Incunabula: With a Revised Check-List of the Incunabula in the Army Medical Library, Washington, D. C. PMID- 19311736 TI - John of Arderne: Medieval English Surgeon. PMID- 19311735 TI - Psychiatry's Part in Preventive Medicine. PMID- 19311737 TI - Report of Dinner Given in Honor of Dr. William H. Welch at The New York Academy of Medicine on April 4, 1930. PMID- 19311738 TI - On the Training of Specialists. PMID- 19311739 TI - Council Accepts Scholarship. PMID- 19311740 TI - Newer Sidelights on the Antiquity and Provenance of Indian Medicine. PMID- 19311742 TI - Report on the Spas of Europe. PMID- 19311741 TI - The Nonvalvular Diseases of the Heart in Middle Life. PMID- 19311743 TI - The Eye in Relation to General Medicine. PMID- 19311744 TI - Specific Sensitiveness as a Cause of Symptoms in Disease. PMID- 19311745 TI - Drug Addiction. PMID- 19311746 TI - Third Annual Graduate Fortnight. PMID- 19311747 TI - Friday Afternoon Lectures, 1980-1931. PMID- 19311748 TI - Can Our Methods of Obstetric Practice be Improved? PMID- 19311749 TI - William and John Hunter-A Study in Contrasts. PMID- 19311750 TI - Suggestions for a National Nomenclature of Diseases. PMID- 19311752 TI - Committee on Medical Education. PMID- 19311751 TI - Blood Transfusion Betterment Association. PMID- 19311754 TI - Scientific Exhibit. PMID- 19311753 TI - Immunity-General and Local. PMID- 19311755 TI - Focal Infection as the Cause of General Disease. PMID- 19311756 TI - Medical Education and Medical Practice. PMID- 19311757 TI - Acute Hematogenous Osteomyelitis. PMID- 19311759 TI - Infections of the Middle Ear. PMID- 19311758 TI - Acute Infections of the Urinary Tract. PMID- 19311760 TI - Infections of the Oral Cavity. PMID- 19311761 TI - Reminiscences of Dr. S. Weir Mitchell. PMID- 19311763 TI - Operative Risks from Infection. PMID- 19311762 TI - Extract from Report of Committee on Professional Standards. PMID- 19311764 TI - Presidential Address. PMID- 19311765 TI - Thomas W. Salmon Memorial. PMID- 19311766 TI - Proposed New Classification of Membership in the Academy. PMID- 19311767 TI - Allocation of Dues. PMID- 19311768 TI - Upper Respiratory Focal Infections in Some of their Regional and General Manifestations. PMID- 19311770 TI - Bacteriemia. PMID- 19311769 TI - Appendicitis. PMID- 19311771 TI - Complications Incident to Inaccurate or Delayed Diagnosis in Suppuration of the Lungs and Pleura. PMID- 19311772 TI - Acute Infections of the Gall Bladder and Biliary Tract. PMID- 19311773 TI - On the Nature of Fits. PMID- 19311775 TI - Standards for Oxygen Therapy Equipment: Committee on Public Health Relations. PMID- 19311774 TI - Extract from the Minutes of the Council in Regard to Dr. Highman. PMID- 19311776 TI - Demonstration of Oxygen Therapy. PMID- 19311777 TI - Death of Mr. John S. Brownne: Resolutions Passed at the Stated Meeting of the Academy Held March 5, 1931. PMID- 19311778 TI - Puerperal Mortality and Its Reduction. PMID- 19311779 TI - Clinical and Pathological Notes on Puerperal Infection. PMID- 19311780 TI - Puerperal Infections and Their Present Therapy. PMID- 19311781 TI - The Therapeutics of Ultraviolet Light. PMID- 19311783 TI - Hermann Michael Biggs Memorial Lecture. PMID- 19311782 TI - Dr. William Beaumont: His Life and Associations in Plattsburgh, N. Y. PMID- 19311785 TI - Bacteriophage as a Treatment in Acute Medical and Surgical Infections. PMID- 19311784 TI - The Relation of the Medical Profession to "Birth Control". PMID- 19311786 TI - Infections of Skin and the Subcutaneous Tissue. PMID- 19311787 TI - John S. Brownne. PMID- 19311788 TI - Serum Therapy. PMID- 19311789 TI - Facts and Fallacies Concerning Foreign Protein and Vaccine Therapy. PMID- 19311791 TI - The Continued Education of the Doctor. PMID- 19311790 TI - Factors Favoring the Onset and Continuation of Rheumatic Fever. PMID- 19311793 TI - Extension to Academy Building. PMID- 19311792 TI - Award of Alexander Cochran Bowen Scholarship. PMID- 19311794 TI - Dr. Paolo De Vecchi. PMID- 19311795 TI - An Epitome of the History of Spanish Medicine. PMID- 19311796 TI - Cancer as a Complication of Skin Diseases. PMID- 19311797 TI - More Common Diseases of the Teeth and Jaws. PMID- 19311798 TI - Russian Medicine Under the Old Regime. PMID- 19311799 TI - The Control of Chronic Diseases. PMID- 19311801 TI - Effect of Air Pollution on Health: Report of the Committee on Public Health Relations of The New York Academy of Medicine. PMID- 19311800 TI - Tropical Diseases and Parasitic Infections in New York City. PMID- 19311802 TI - An Outline of the History of the Circulatory System. PMID- 19311803 TI - Recent Contributions in Diseases of the Thyroid and Diabetes. PMID- 19311804 TI - Principles Governing Contact of Physicians with the Public Science and Cultism. PMID- 19311805 TI - Science and Cultism. PMID- 19311807 TI - The Romantic Episode in the History of German Medicine. PMID- 19311806 TI - International Hospital Association. PMID- 19311808 TI - Address of Welcome. PMID- 19311809 TI - Introductory Remarks. PMID- 19311810 TI - The Wesley M. Carpenter Lecture: Muscular Pain in Intermittent Claudication. PMID- 19311812 TI - Herbals and Bestiaries. PMID- 19311811 TI - Resolutions on Medicinal Alcohol: Preamble and Resolutions Adopted by the Council on March 25, 1931 and Amendments. PMID- 19311813 TI - Jean-Dominique Larrey and the "Academie De New York,". PMID- 19311815 TI - Diet in Disease. PMID- 19311814 TI - Meningococcus Infections Including Meningitis. PMID- 19311816 TI - Address. PMID- 19311818 TI - Report on Necropsies: Prepared by the Joint Committee representing. PMID- 19311817 TI - Remarks. PMID- 19311819 TI - Poliomyelitis in New York City: Progress Report of the Sub-committee. PMID- 19311820 TI - FRONTISPIECE. PMID- 19311821 TI - A TRIBUTE. PMID- 19311822 TI - The University and the Medical Profession. PMID- 19311823 TI - Pure Air and Clean Streets Exhibit. PMID- 19311824 TI - Arteriosclerosis. PMID- 19311825 TI - A List of Writings on the Cardiovascular System Exhibited during the Fortnight. PMID- 19311826 TI - Review of Academy Activities. PMID- 19311827 TI - Library Notes. PMID- 19311828 TI - Cutaneous Manifestations of Systemic Diseases. PMID- 19311830 TI - Unethical Practice-Ambulance Fees. PMID- 19311829 TI - In Regard to Prescribing Alcoholic Liquors. PMID- 19311831 TI - Publication of the Salmon Memorial Lectures. PMID- 19311832 TI - Psychoneuroses and Neuroses and the Heart. PMID- 19311833 TI - Malignancies of the Colon: The Medical Aspect. PMID- 19311834 TI - The Roentgenologic Manifestations and Differential Diagnosis of Carcinoma of the Colon. PMID- 19311835 TI - Surgical Aspects of Carcinoma of the Colon. PMID- 19311836 TI - The First Authentic Periodical of Medical History. PMID- 19311837 TI - The Treatment of Neurosyphilis. PMID- 19311838 TI - Cardiovascular Syphilis. PMID- 19311839 TI - The Pathology of Syphilis: With Special Reference to the Development of Luetic Aortitis. PMID- 19311840 TI - General Discussion. PMID- 19311841 TI - Fontenelle as a Popularizer of Science. PMID- 19311842 TI - Point of View of Human Relationships. PMID- 19311843 TI - Psychoneuroses in Children. PMID- 19311844 TI - Certain Pediatric Doubts About Modern Psychiatry. PMID- 19311845 TI - Report on Housing: Committee on Public Health Relations. PMID- 19311846 TI - Report of Museum Committee. PMID- 19311847 TI - The Literary Tradition in English Medicine, (With Sidelights on Medicine in English Literature). PMID- 19311848 TI - Robert Koch. PMID- 19311849 TI - Medical Geography and Geographic Medicine. PMID- 19311850 TI - Report of Study of Administration of Convalescent Serum in the Treatment of Poliomyelitis. PMID- 19311851 TI - Friday Afternoon Lectures. PMID- 19311852 TI - Address of Welcome. PMID- 19311853 TI - The Historical Development of the Pathology and Therapy of Cancer. PMID- 19311854 TI - The Wesley M. Carpenter Lecture: Fundamental Research in Cancer. PMID- 19311855 TI - Group Purchase of Hospital Care. PMID- 19311856 TI - Some Aspects of Cancer Mortality. PMID- 19311857 TI - Summary of Results that can be Obtained by Irradiation in Cancer. PMID- 19311858 TI - Migraine and Its Treatment. PMID- 19311859 TI - Some Facts Concerning Tumors of the Brain. PMID- 19311861 TI - Simple Achlorhydric Anemia. PMID- 19311860 TI - Gastro-Intestinal Manifestations of Allergy. PMID- 19311862 TI - On Sydenham's View of Causation in the Light of Seventeenth Century Thought. PMID- 19311864 TI - Address of the Incoming President: The Academy: Its Relations to the Art and Practice of Medicine in New York. PMID- 19311863 TI - Address of the Retiring President: Problems Facing the Academy. PMID- 19311865 TI - Funds for Research in Psychiatry. PMID- 19311866 TI - Rise of the Hospital Idea. PMID- 19311867 TI - The Psychiatric Approach of the Practitioner to His Patients. PMID- 19311868 TI - Review of "Poliomyelitis,": A Survey Made Possible By a Grant From The International Committee For The Study Of Infantile Paralysis. PMID- 19311869 TI - Cost of Current Medical German Periodicals. PMID- 19311871 TI - Hermann Michael Biggs Memorial Lecture. PMID- 19311870 TI - Graduate Fortnight Exhibit Illustrating the History of the Evolution of the Knowledge of Cancer. PMID- 19311872 TI - The Medical Literature of France. PMID- 19311873 TI - Fee-Splitting. Report of the Committee on Professional Standards: Approved by the Council March 22, 1933. PMID- 19311874 TI - Treating the Pneumonia Patient. PMID- 19311875 TI - The Health and Welfare Resources of the City for the Physicians. PMID- 19311876 TI - The Skin as a Functional Organ of the Body. PMID- 19311877 TI - Recent Developments in the Study of Tuberculosis of Interest to Physicians. PMID- 19311879 TI - The Medical Information Bureau of The New York Academy of Medicine. PMID- 19311878 TI - Scurvy-"The Plague of the Sea and the Spoyle of Mariners". PMID- 19311880 TI - Friday Afternoon Lectures. PMID- 19311882 TI - Diabetes Mellitus-Problems of Present Day Treatment. PMID- 19311881 TI - The Prognosis of Heart Disease. PMID- 19311883 TI - Public Health Problems. PMID- 19311884 TI - Heredity and Prevention. PMID- 19311885 TI - Clinical Phases. PMID- 19311886 TI - Recent Trends in Diabetes Mortality. PMID- 19311887 TI - William Beaumont as an Army Officer. PMID- 19311888 TI - The Beaumont-St. Martin Contract and the Descendants of Dr. Beaumont. PMID- 19311889 TI - The Book of William Beaumont after One Hundred Years. PMID- 19311890 TI - George David Stewart, A Tribute. PMID- 19311891 TI - Committee on Fellowship. PMID- 19311892 TI - Introductory Remarks. PMID- 19311893 TI - On Quackery as a Reversion to Primitive Medicine. PMID- 19311895 TI - Report on the Sixth Annual Graduate Fortnight. PMID- 19311894 TI - The Wesley M. Carpenter Lecture: The Influence of the Diencephalon and Hypophysis Upon General Autonomic Function. PMID- 19311896 TI - The Physician's Profession Through the Ages. PMID- 19311897 TI - Introductory Address to the Sixth Annual Graduate Fortnight of The New York Academy of Medicine. PMID- 19311898 TI - Director ad interim. PMID- 19311899 TI - Total Energy Exchange in Relation to Clinical Medicine. PMID- 19311900 TI - ANNIVERSARY DISCOURSE: "Medicine and Modern Sociological Trends". PMID- 19311901 TI - The Metabolism of Fever, with Special Reference to Diabetic Hyperpyrexia. PMID- 19311902 TI - Recent Action of the Council Regarding Fee-Splitting. PMID- 19311903 TI - Alfred Fabian Hess. PMID- 19311904 TI - Chapman, Charles Francis. PMID- 19311905 TI - Moschcowitz, Alexis Victor. PMID- 19311906 TI - Franken, Sigmund Walter Anthony. PMID- 19311907 TI - Williams, Linsly Rudd. PMID- 19311908 TI - Bureau of Clinical Information. PMID- 19311910 TI - Metabolism in Hyperthyroidism and Hypothyroidism. PMID- 19311909 TI - Linsly Rudd Williams. PMID- 19311911 TI - Surgical Treatment of Hyperthyroidism. PMID- 19311912 TI - Mineral Metabolism. PMID- 19311913 TI - Acidosis and Alkalosis. PMID- 19311914 TI - Dehydration and Medical Shock. PMID- 19311915 TI - Fluid Distribution and Edema. PMID- 19311916 TI - Congenital Anomalies of Metabolism with Special Reference to Cystinuria and Myopathies. PMID- 19311917 TI - Gout. PMID- 19311918 TI - Clinical and Biologic Considerations of Obesity and Certain Allied Conditions. PMID- 19311919 TI - A Critical Estimate of the Value of Laboratory Procedures in Disorders of Metabolism. PMID- 19311920 TI - Clinical Observations and Laboratory Investigations on the 1933 Epidemic of Encephalitis in St. Louis. PMID- 19311921 TI - The Thomas W. Salmon Memorial Committee Report and Presentation of Portrait of the Late Dr. Thomas W. Salmon. PMID- 19311922 TI - General review of our present knowledge of the vitamins. PMID- 19311923 TI - The effects of moderate deficiency of vitamins. PMID- 19311925 TI - Charles Darwin and the history of the early use of digitalis. PMID- 19311924 TI - Non-diabetic ketosis in children. PMID- 19311926 TI - Committee on Public Health Relations Report on Vitamin D Milk. PMID- 19311927 TI - The Medical History of Robert Schumann and His Family. PMID- 19311928 TI - Hyperparathyroidism and Its Relationship to Diseases of Bone. PMID- 19311929 TI - Metabolic Disturbances in Relation to the Teeth. PMID- 19311930 TI - Transvaluations and Deflations in the History of Medicine and Its Teaching. PMID- 19311931 TI - The Causes and Prevention of Blindness. PMID- 19311933 TI - Report of the Council, Charges Against Dr. Fred H. Albee. PMID- 19311932 TI - A Note on Narcotics in Ancient Greece and in Ancient China. PMID- 19311934 TI - History of Gastro-Enterology. PMID- 19311936 TI - The Opening of the Seventh Annual Graduate Fortnight. PMID- 19311935 TI - The Wesley M. Carpenter Lecture: The Applied Physiology of the Gastro-Intestinal Innervation. PMID- 19311938 TI - Lusk, William Chittenden. PMID- 19311937 TI - Ramon y Cajal, Santiago. PMID- 19311939 TI - Life as an Occupational Disease. PMID- 19311941 TI - Early 19th Century Dermatology and the Brothers Mahon. PMID- 19311940 TI - Historical Points of Interest on the Mode of Action and Ill Effects of Mercury. PMID- 19311943 TI - Address of the Retiring President: Standards-Achievements. PMID- 19311942 TI - Photograph of J. Marion Sims. PMID- 19311944 TI - Discussion of the Workmen's Compensation Law in New York: Its Experience And Its Implications. PMID- 19311945 TI - Medical Care and Hospitalization of the Indigent Sick. PMID- 19311946 TI - Report on the Seventh Annual Graduate Fortnight. PMID- 19311947 TI - Address of the Incoming President: Problems Facing Medicine. PMID- 19311948 TI - ANNIVERSARY DISCOURSE: The Medical Profession in the New Age. PMID- 19311949 TI - Exhibition in the Library: The Development of the Stethoscope. PMID- 19311950 TI - The Doctor and the State. PMID- 19311951 TI - Felice Fontana: A Forgotten Physiologist of the Trentino. PMID- 19311953 TI - The Modern Attitude Toward Traumatic Cancer. PMID- 19311952 TI - The Problem of Acute Appendicitis in New York City. PMID- 19311954 TI - Amendments to By-Laws. PMID- 19311955 TI - Fielding H. Garrison. PMID- 19311956 TI - The principles governing the radiation therapy of cancer. PMID- 19311957 TI - Benign and malignant lesions of the female reproductive tract. PMID- 19311958 TI - Present status of cutaneous x-ray, grenz-ray and radium therapy. PMID- 19311959 TI - The roentgen ray treatment of tumors of the brain and skull. PMID- 19311960 TI - A Skin Test to Suggest the Diagnosis of Recovered Typhus and Thrombo-angiitis. PMID- 19311961 TI - General Considerations. PMID- 19311962 TI - The Treatment of the Patient with Angina Pectoris. PMID- 19311964 TI - John Hughlings Jackson. PMID- 19311963 TI - An Evaluation of the Surgical Treatment of Angina Pectoris. PMID- 19311965 TI - Letters of Samuel Hahnemann: Presented to the Library. PMID- 19311967 TI - Public Responsibility for Public and Personal Health: The Biggs Health Center Plan of 1920 in Retrospect. PMID- 19311966 TI - The Earliest Modern Law for the Regulation of the Practice of Medicine. PMID- 19311969 TI - The Medical Man and the Witch Towards the Close of the Sixteenth Century. PMID- 19311968 TI - Observations on Tuberculous Abscess of the Chest Wall. PMID- 19311970 TI - The Development of the Stethoscope: An Exhibition Showing the Work of Laennec and His Successors Held in the Library February 13 to March 29, 1935. PMID- 19311971 TI - The Wesley M. Carpenter Lecture: Atelectasis, Massive Collapse and Related Post Operative Conditions. PMID- 19311972 TI - The Graduate Fortnight of 1935. PMID- 19311973 TI - ANNIVERSARY DISCOURSE: "ASPECTS OF A PHILOSOPHY OF GOVERNMENT IN A SICK WORLD". PMID- 19311975 TI - Claude Tardi, An Early Advocate of Direct Transfusion of Human Blood. PMID- 19311974 TI - James De Berty Trudeau: Artist, Soldier, Physician: Examples of His Work. PMID- 19311976 TI - Brief History of Organized Medicine. PMID- 19311977 TI - The Relations of the Academy to the Public and the Profession. PMID- 19311978 TI - George David Stewart, Man and Poet. PMID- 19311979 TI - Resolution of the Council. PMID- 19311980 TI - Alfred Grotjahn, Founder of Social Hygiene. PMID- 19311981 TI - The Domestic Relations Court: Report of the Subcommittee appointed by The Committee on Public Health Relations of The New York Academy of Medicine. PMID- 19311983 TI - Marion Sims and the Origin of Modern Gynecology. PMID- 19311982 TI - Beethoven, from an Otologist's Viewpoint. PMID- 19311984 TI - Introductory Remarks of the Chairman. PMID- 19311985 TI - Dr. Harrison Stanford Martlan-An Appreciation. PMID- 19311986 TI - Introduction of George Hoyt Whipple. PMID- 19311987 TI - Problems in Anemia. PMID- 19311988 TI - Resolutions of the Council on the deaths of Dr. Arthur B. Duel and Dr. Harlow Brooks. PMID- 19311990 TI - The Physiology of Vesalius. PMID- 19311989 TI - A Reading from Vesalius. PMID- 19311991 TI - The Eugenic Outlook in Preventive Medicine. PMID- 19311992 TI - The Improvements in the Ability of the Medical Profession to Treat Cancer. PMID- 19311993 TI - Role of the Nestorians as the Connecting Link between Greek and Arabic Medicine. PMID- 19311994 TI - The Initial Symptoms and Early Diagnosis of Tumor of the Brain. PMID- 19311996 TI - Address of Dr. D. Bryson Delavan. PMID- 19311995 TI - Carcinoma of the Colon and Rectum. PMID- 19311997 TI - Advice of Nathan Smith on the Conduct of an Accoucheur. PMID- 19311998 TI - New York Physicians Registered in the Office of the County Clerk in 1806. PMID- 19311999 TI - Certain Old American Medical Works. PMID- 19312000 TI - Frans Cornelis Donders-Dutch Physiologist and Ophthalmologist. PMID- 19312001 TI - Report of Committee on Lectures to the Laity. PMID- 19312002 TI - Opening Remarks. PMID- 19312003 TI - The Wesley M. Carpenter Lecture: "Historical Background of Industrial and Occupational Diseases". PMID- 19312004 TI - Shock and Hemorrhage. PMID- 19312005 TI - Report on the Ninth Annual Graduate Fortnight. PMID- 19312007 TI - Medico-Legal Aspects of Industrial Poisonings. PMID- 19312006 TI - Relation of Trauma to Pulmonary Disease. PMID- 19312008 TI - A Case of Pericarditis with Effusion: Studies of Venous Pressure Changes. PMID- 19312009 TI - Change in Advertising Policy. PMID- 19312010 TI - Recurrent Pericardial Effusion of Unknown Etiology: Report of a Case with Studies of the Circulation and of Absorption from the Pericardical Cavity. PMID- 19312011 TI - ANNIVERSARY DISCOURSE: The Professions in World Turmoil. PMID- 19312012 TI - The Medical Problem in First Aid. PMID- 19312013 TI - Annual Postgraduate Institute of the Philadelphia County Medical Society. PMID- 19312015 TI - Thomas William Salmon Memorial Lectures. PMID- 19312014 TI - Recent Activities of the New York Academy of Medicine. PMID- 19312016 TI - Ideals in Medicine. PMID- 19312017 TI - Medicine and the State. PMID- 19312018 TI - The Origin and Nature of the Hypoglycemic Therapy of the Psychoses. PMID- 19312019 TI - Some Functions of the Hypothalamus: Harvey Lecture, December 17, 1936. PMID- 19312021 TI - The Academy Scholarship. PMID- 19312020 TI - The Investigation of Intermediary Metabolism with the Aid of Heavy Hydrogen: Harvey Lecture, January 21, 1937. PMID- 19312022 TI - The Bearing of the Results of Recent Studies in Nutrition on Health and on Length of Life. PMID- 19312023 TI - The Control of Excitation in the Nervous System. PMID- 19312024 TI - Rabelais as a Physician. PMID- 19312025 TI - Transmission of Nervous Effects by Acetylcholine: Harvey Lecture, May 20, 1937. PMID- 19312027 TI - A Review of "American Medicine". PMID- 19312026 TI - Chemical Warfare as Developed During the World War-Probable Future Development. PMID- 19312028 TI - The Scientific Work of the Health Organization of the League of Nations. PMID- 19312029 TI - Count Cagliostro-An Excursion Into Eighteenth Century Charlatanism. PMID- 19312030 TI - Modern Concepts of Anemia from the Clinical Standpoint. PMID- 19312032 TI - Jonathan Hutchinson: The Last of the Multispecialists. PMID- 19312031 TI - Medical Explorers of Arabia. PMID- 19312033 TI - Community Provision for the Serum Treatment of Pneumococcic Pneumonias. PMID- 19312035 TI - Postgraduate Medical Education-with Especial Reference to Present Maternal Mortality Rates in the United States. PMID- 19312034 TI - Robin and John Adair. PMID- 19312036 TI - Address of Welcome to the Tenth Annual Graduate Fortnight. PMID- 19312038 TI - The Nature of Hypertension. PMID- 19312037 TI - The Nephroses. PMID- 19312040 TI - New Technique in the Surgical Treatment of Severe and Progressive Deafness from Otosclerosis. PMID- 19312039 TI - The Factual Background for the Treatment of Progressive Deafness from Otosclerosis. PMID- 19312041 TI - Evaluation of the Surgical Treatment of Hypertension. PMID- 19312042 TI - Prerenal Azotemia and the Pathology of Renal Blood Flow. PMID- 19312044 TI - Editorial Announcement. PMID- 19312043 TI - The Nature of the Visual Process: Harvey Lecture, October 21, 1937. PMID- 19312045 TI - Physiology of the Kidney: The Wesley M. Carpenter Lecture. PMID- 19312047 TI - Clinical Aspects of Nephritis. PMID- 19312046 TI - The Nature of Glomerulonephritis. PMID- 19312048 TI - Calculous Disease in the Urinary Tract: The Formation of Stone. PMID- 19312049 TI - Clinical Aspects of Calculus Disease. PMID- 19312050 TI - Edema and Its Treatment. PMID- 19312051 TI - Management of Tumors of the Kidney and Ureter. PMID- 19312052 TI - The Significance of The N. Y. Academy of Medicine: Presidential Address. PMID- 19312053 TI - Milestones in Medicine. PMID- 19312055 TI - The Conception of Cancer Before and After Johannes Muller. PMID- 19312054 TI - The Pasteur-Meyerhof Reaction in Muscle Metabolism: Harvey Lecture, November 18, 1937. PMID- 19312056 TI - Emotional Factors in Hypertension. PMID- 19312057 TI - The Treatment of Acute Glomerulonephritis in Children. PMID- 19312058 TI - The Functional Significance of the Lymphatic System: Harvey Lecture, December 16, 1937. PMID- 19312060 TI - Pathological Physiology of Bladder Neck Obstruction. PMID- 19312059 TI - Radiotherapy of Tumors of the Urinary Tract. PMID- 19312061 TI - The Meaning of Medical Research. PMID- 19312062 TI - Transfers of Water and Solutes in the Body: Harvey Lecture, January 20, 1938. PMID- 19312063 TI - Cancer of the Gastrointestinal Canal: The Bulkley Lecture. PMID- 19312064 TI - Clinical Aspects of Hypertension including Malignant Hypertension. PMID- 19312065 TI - The Use of Convalescent Scarlet Fever and Measles Sera in Prophylaxis and Therapy. PMID- 19312066 TI - Viruses and Virus Diseases: Twentieth Century Version of the De Novo Origin of Infectious Agents and Its Significance in Relation to the Control of Disease. PMID- 19312067 TI - Isolation and Properties of Tobacco Mosaic and Other Virus Proteins: Harvey Lecture, March 17, 1938. PMID- 19312068 TI - The Vascular and Renal Complications of Pregnancy. PMID- 19312069 TI - The Treatment of Hemolytic Streptococcus Infections and the Newer Applications of Sulphanilamide. PMID- 19312070 TI - The Modern Treatment of Diabetes. PMID- 19312071 TI - Hieronymus Muenzer and Other Fifteenth Century Bibliophiles. PMID- 19312072 TI - Experimental Hypertension Induced by Renal Ischemia: Harvey Lecture, May 19, 1938. PMID- 19312073 TI - Endoscopic Prostatic Resection. PMID- 19312074 TI - The Specific Prevention of Diphtheria: Further Observations and Inquiries. PMID- 19312075 TI - Studies on the Cortical Representation of Somatic Sensibility: Harvey Lecture, February 17, 1938. PMID- 19312076 TI - The Present Status of Gynecologic Endocrine Therapy. PMID- 19312077 TI - The Pathological Responses to Vitamin Deficiencies. PMID- 19312078 TI - Our Vitamin B1 Supply in Relation to Human Needs. PMID- 19312079 TI - The Chemistry and Biology of Male Sex Hormones. PMID- 19312081 TI - Address of Welcome to the Eleventh Annual Graduate Fortnight. PMID- 19312080 TI - The Management of Hypertension. PMID- 19312082 TI - Remarks on the Differential Diagnosis and Treatment of Pernicious Anemia. PMID- 19312083 TI - The Neutropenic Diseases. PMID- 19312084 TI - Recent Advances in Knowledge of Some of the Common Diseases of Childhood. PMID- 19312085 TI - Some Aspects of the Intermediary Metabolism of the Steroid Hormones: Harvey Lecture, October 20, 1938. PMID- 19312086 TI - Hemophilia: The Wesley M. Carpenter Lecture. PMID- 19312088 TI - Present Status of Serum Therapy in Pneumonia. PMID- 19312087 TI - Infectious Mononucleosis. PMID- 19312089 TI - Treatment of Pneumonia with Antipneumococcal Rabbit Serum. PMID- 19312090 TI - The Significance of the Albumin Fraction of Serum: Harvey Lecture, November 17, 1938. PMID- 19312091 TI - General Pathology of Lymphosarcoma. PMID- 19312092 TI - Address of the Retiring President. PMID- 19312093 TI - Presidential Address. PMID- 19312094 TI - Heat Loss from the Human Body: Harvey Lecture, December 15, 1938. PMID- 19312096 TI - Hemolytic Jaundice: Its Diagnosis, Behavior and Treatment. PMID- 19312095 TI - The Medical-Surgical Splenopathies. PMID- 19312097 TI - Congestive Splenomegaly. PMID- 19312099 TI - Diagnostic Significance of Changes in Leukocytes. PMID- 19312098 TI - Diagnostic and Therapeutic Considerations in the Management of Idiopathic Thrombocytopenic Purpura. PMID- 19312100 TI - Preliminary Report of the Use of Sulfapyridine in the Treatment of Pneumonia. PMID- 19312101 TI - Preliminary Report of the Use of Sulfapyridine in the Treatment of Pneumonia. PMID- 19312102 TI - Preliminary Report of the Use of Sulfapyridine in the Treatment of Pneumonia: The Absorption, Acetylation and Excretion of Sulfapyridine. PMID- 19312103 TI - The Treatment of Depersonalization. PMID- 19312104 TI - Present Status of Protamine Insulin. PMID- 19312105 TI - The Diagnostic Significance of Changes in the Red Cells. PMID- 19312107 TI - Chronic Gastritis. PMID- 19312106 TI - Polycythemia. PMID- 19312108 TI - Modern Treatment of Schizophrenia. PMID- 19312109 TI - Genic and Hormonal Factors in Biological Processes: Harvey Lecture, April 20, 1939. PMID- 19312110 TI - Clinical and Pathological Aspects of Acute Leukemia. PMID- 19312111 TI - Chronic Gastritis: Clinical Aspects. PMID- 19312112 TI - Vitamin A with Special Reference to Therapy. PMID- 19312113 TI - The Significance to Medicine of Present Population Trends: The Hermann Michael Biggs Memorial Lecture. PMID- 19312114 TI - Irradiation in the Lymphomatoid Diseases. PMID- 19312115 TI - Biological Oxidation and Vitamins: Harvey Lecture, May 18, 1939. PMID- 19312116 TI - The Diagnosis, Treatment, and Prevention of Vitamin B1 Deficiency. PMID- 19312117 TI - Observations on the Pathology of Rickets with Particular Reference to the Changes at the Cartilage-shaft Junctions of the Growing Bones: Harvey Lecture, February 16, 1939. PMID- 19312118 TI - The Therapeutic Use of Vitamin C. PMID- 19312119 TI - The Use of Tetanus Toxoid in Private Practice. PMID- 19312121 TI - The Role of The New York Academy of Medicine in the Development of The American Museum of Health. PMID- 19312120 TI - The Influence of Emotional Factors upon Physiological and Pathological Processes. PMID- 19312122 TI - Some Difficulties in the Use of the Insulins in Diabetic Practice. PMID- 19312124 TI - Pathogenesis and Present Day Treatment of Urinary Infections. PMID- 19312123 TI - The Treatment of Postabortal and Postpartum Sepsis, with Special Consideration of Sulfanilamide and Allied Drugs. PMID- 19312125 TI - Proteins as Chemical Substances and as Biological Components. PMID- 19312126 TI - On the Origin and Developmental Potentialities of Blood Cells. PMID- 19312127 TI - The Curious Career of Typhoid Mary. PMID- 19312129 TI - Some Comments on Arteriosclerosis in Wild Mammals and Birds. PMID- 19312128 TI - Distribution of Enzymes in Tissue and Cells: Harvey Lecture, March 16, 1939. PMID- 19312130 TI - Psychological Study of the Migrainous Syndrome. PMID- 19312131 TI - Address of Welcome to the Twelfth Graduate Fortnight. PMID- 19312132 TI - Hypothalamic-Pituitary Syndromes. PMID- 19312133 TI - Physiology and Principal Interrelations of the Thyroid Gland. PMID- 19312134 TI - Lectures to the Laity. PMID- 19312135 TI - Hypothyroidism: Diagnosis and Treatment. PMID- 19312136 TI - The Modern Treatment of Pyogenic Osteomyelitis. PMID- 19312137 TI - Otitis Media and Its Extensions. PMID- 19312138 TI - The Adrenal Medulla. PMID- 19312139 TI - Obstetrics at the New York Almshouse and at Bellevue Hospital. PMID- 19312140 TI - Convalescence of Old-Age Patients. PMID- 19312142 TI - Puberty, Menstruation, Pregnancy. PMID- 19312141 TI - Treatment of Gonadal Hypofunction. PMID- 19312143 TI - The Physiology and Psychology of Convalescence. PMID- 19312144 TI - Presidential Address. PMID- 19312145 TI - Physiology of the Testes and Therapeutic Application of Male Hormone. PMID- 19312146 TI - The Physiology of the Ovaries. PMID- 19312147 TI - The Biological Significance of Nicotinic Acid: Harvey Lecture, November 16, 1939. PMID- 19312149 TI - Chemotherapy of Pneumonia. PMID- 19312148 TI - Chemotherapy with the Sulfonamide Derivatives: General Principles. PMID- 19312150 TI - A Consideration of Some of the Toxic Effects of Sulfonamide Compounds, Particularly Sulfapyridine. PMID- 19312151 TI - Hyperpituitarism and Hypopituitarism. PMID- 19312152 TI - Hypertension-The Problem, The Study, The Future. PMID- 19312153 TI - The Medical Management of Hyperthyroidism. PMID- 19312154 TI - Hyperparathyroidism. PMID- 19312155 TI - Adrenal Insufficiency. PMID- 19312156 TI - The Cushing Syndrome; Neoplasms of the Adrenal Gland. PMID- 19312157 TI - Surgical Considerations in the Treatment of Chronic Lymphedema and of Varicose Veins. PMID- 19312158 TI - Mechanism of Allergy. PMID- 19312159 TI - Allergy in Childhood. PMID- 19312161 TI - Heart Disease-a World Problem. PMID- 19312160 TI - Analysis of Maternal Deaths and Hospital Obstetrical Statistics in New York County. PMID- 19312162 TI - The Menopause. PMID- 19312164 TI - Purposes, Function and Use of Standard Classified Nomenclature of Disease. PMID- 19312163 TI - Clinical Aspects of Rheumatic Fever in Children. PMID- 19312165 TI - The Adaptation of the Standard Classified Nomenclature of Disease to Hospital Morbidity Reports. PMID- 19312166 TI - Features Which Suggest Public Health Consideration of Rheumatic Fever. PMID- 19312167 TI - Clinical Aspects of Rheumatic Fever in Adults. PMID- 19312168 TI - Management of the Anemias in Infancy and Childhood. PMID- 19312169 TI - Convalescence in Coronary Disease. PMID- 19312170 TI - The Problem of Aging. PMID- 19312171 TI - Histaminase: Physiologic Effects on Man and Its Therapeutic Value in Medicine. PMID- 19312172 TI - The Medical Management of Disorders of the Biliary Tract. PMID- 19312173 TI - Aging of the Cardiovascular System. PMID- 19312174 TI - Histamine in Anaphylaxis and Allergy. PMID- 19312175 TI - An American Precursor of Freud. PMID- 19312176 TI - The Pathological Aspects of Rheumatic Fever. PMID- 19312177 TI - Treatment of Mental Disease in France at the End of the Eighteenth Century. PMID- 19312178 TI - A Review of the Maternity Statistics of New York City for the Year 1939. PMID- 19312180 TI - The Clinical Use of Sulfanilamide and Its Derivatives in the Treatment and Prophylaxis of Certain Infections: The Wesley M. Carpenter Lecture. PMID- 19312179 TI - Experimental Basis of Chemotherapy in the Treatment of Bacterial Infections: The Ludwig Kast Lecture. PMID- 19312181 TI - Electrophoretic Analysis and the Constitution of Native Fluids: Harvey Lecture, October 19, 1939. PMID- 19312182 TI - The Blood Plasma for Great Britain Project. PMID- 19312183 TI - The Sex Hormones and the Endocrine Balance: The Middleton Goldsmith Lecture of The New York Pathological Society. PMID- 19312184 TI - Gonorrhea in the Male. PMID- 19312185 TI - Current Trends in Diagnosis of Renal Tuberculosis. PMID- 19312186 TI - The Treatment of Renal Tuberculosis. PMID- 19312187 TI - Gonococcal Infections of Women. PMID- 19312188 TI - Treatment of Bacterial Meningitis. PMID- 19312190 TI - Massive Dose Chemotherapy by the Intravenous Drip Method. PMID- 19312189 TI - The Newer Knowledge of Vitamin K. PMID- 19312191 TI - Humanism and Science: The Linsly R. Williams Memorial Lecture. PMID- 19312193 TI - Medical Aspects of Arterial Hypertension. PMID- 19312192 TI - Inaugural Address of the President of The New York Academy of Medicine: What Is The New York Academy of Medicine? PMID- 19312194 TI - Pneumococcal and "Virus" Pneumonia. PMID- 19312195 TI - Newer Knowledge of the Vitamin B-Complex. PMID- 19312197 TI - The Prevention and Treatment of Infection in Wounds both Operative and Accidental. PMID- 19312196 TI - Osteomyelitis and Pyogenic Infections of Joints. PMID- 19312198 TI - Virus Infections. PMID- 19312199 TI - Poliomyelitis. PMID- 19312201 TI - The Rickettsial Diseases, with Special Reference to those of Importance Along the Atlantic Seaboard. PMID- 19312200 TI - Epidemic Influenza. PMID- 19312202 TI - Studies on Choriomeningitis and Poliomyelitis: Harvey Lecture, October 31, 1940. PMID- 19312204 TI - Newer Conceptions of Postinfectious and Related Forms of Encephalitis. PMID- 19312203 TI - Oliver Wendell Holmes: A Precursor of Freud. PMID- 19312205 TI - Thrombophlebitis. PMID- 19312206 TI - Newer Knowledge of Blood Transfusions. PMID- 19312207 TI - Utilization of Selective Microbial Agents in the Study of Biological Problems: Harvey Lecture, March 21, 1940. PMID- 19312208 TI - The Present Status of the Treatment of Subacute Bacterial Endocarditis. PMID- 19312209 TI - Treatment of Infections by Methods other than Chemotherapy. PMID- 19312210 TI - Infections of the Middle Ear and Nasal Sinuses. PMID- 19312211 TI - A Critical Evaluation of the Results of Routine Conservative Treatment of Syphilis. PMID- 19312213 TI - Progress in the Preoperative and Postoperative Care of Patients with Lesions of the Biliary Tract. PMID- 19312212 TI - Epidemic Encephalitis. PMID- 19312214 TI - The Chemical and Physiological Properties, and Clinical Uses of the Corpus Luteum Hormone, Progesterone. PMID- 19312215 TI - Chemical, Physiological and Clinical Aspects of the Androgens. PMID- 19312216 TI - Some Aspects of the Common Contagious Diseases. PMID- 19312217 TI - The Marriage of Medicine and Civil Government. PMID- 19312218 TI - Puerperal Infection: A consideration of the relationship of patholgical changes and bacteriological findings to prevention and treatment. PMID- 19312220 TI - Dental Foci of Infection. PMID- 19312219 TI - Brucellosis. PMID- 19312221 TI - Lymphogranuloma Venereum. PMID- 19312222 TI - Surgical and Non-Surgical Treatment of the Prostate Gland. PMID- 19312223 TI - Infections of the Mouth, Pharynx and Upper Respiratory Tract. PMID- 19312224 TI - The Doctor in Court. PMID- 19312225 TI - The Doctor in Court. PMID- 19312226 TI - The Doctor in Court. PMID- 19312227 TI - Newer Surgery of the Heart and Large Vessels: Medical Aspects. PMID- 19312228 TI - Morphological and Functional Alterations of the Coronary Circulation: Harvey Lecture, April 18, 1940. PMID- 19312229 TI - Effort, Trauma, Occupation and Compensation in Heart Disease. PMID- 19312230 TI - Heparin and Thrombosis: Harvey Lecture, November 28, 1940. PMID- 19312231 TI - The Steering Committee and Academy Financing. PMID- 19312232 TI - Carcinoma of the Stomach Emphasizing Some of the Misconceptions of the Disease. PMID- 19312234 TI - Treatment of Disorders of the Menopause. PMID- 19312233 TI - Acute Surgical Conditions of the Abdomen in Children. PMID- 19312235 TI - Medical Testimony in Personal Injury Action. PMID- 19312236 TI - The Evolution of Clinical Sphygmomanometry. PMID- 19312237 TI - Arteriosclerosis. PMID- 19312238 TI - Coronary Insufficiency: Observations on Diagnosis and Treatment. PMID- 19312239 TI - The Psychopathology of Psychopathic Personalities. PMID- 19312240 TI - The Influence of Climate and Geography on Health. PMID- 19312241 TI - The Doctor in Court. PMID- 19312242 TI - The Founding of the New York Laryngological Society. PMID- 19312243 TI - Department of Health Notice. PMID- 19312244 TI - Heart Failure: The Wesley M. Carpenter Lecture. PMID- 19312245 TI - Basic Hemodynamic Principles Essential to Interpretation of Cardiovascular Disorders: The Ludwig Kast Lecture. PMID- 19312246 TI - Arteriosclerosis: Social Significance and Recent Advances in Treatment. PMID- 19312248 TI - The History of Fever Therapy in the Treatment of Disease. PMID- 19312247 TI - Recent Studies in the Production of Cancer by Chemical Compounds; the Conditioned Deficiency as a Mechanism: The Bulkley Lectuer. PMID- 19312249 TI - Oxygen Supply Systems for Military Flying. PMID- 19312251 TI - A Note on War Psychiatry. PMID- 19312250 TI - Some Recent Advances in Drug Therapy. PMID- 19312252 TI - Friedrich August Kekule: Architect of Atoms. PMID- 19312253 TI - The Inter-America Division of the Academy. PMID- 19312254 TI - Influence of Extrinsic Factors on the Coronary Flow and Clinical Course of Heart Disease. PMID- 19312255 TI - Pulmonary Congestion and Edema. PMID- 19312256 TI - Thrombo-Angiitis Obliterans. PMID- 19312257 TI - Effects of Renal Extract on Hypertension. PMID- 19312258 TI - The Use of Insulin in its Various Forms in the Treatment of Diabetes. PMID- 19312259 TI - Diabetes in Pregnancy. PMID- 19312260 TI - Determining Factors in the End-Results Following War Wounds and Compound Fractures. PMID- 19312261 TI - Evaluation of Drugs Used in the Treatment of Cardiovascular Diseases. PMID- 19312262 TI - Treatment of Leukemia by Radioactive Phosphorus. PMID- 19312263 TI - The Adrenal Cortex and Electrolyte Behavior: Harvey Lecture, December 18, 1941. PMID- 19312264 TI - Walter Lindsay Niles, 1878-1942: An Appreciation. PMID- 19312265 TI - Samuel W. Lambert, 1859-1942. PMID- 19312266 TI - Some Chemical Changes in the Myocardium Accompanying Heart Failure. PMID- 19312267 TI - The Treatment of Gout. PMID- 19312268 TI - Tropical Medicine in United States Military History. PMID- 19312269 TI - Effect of Tonsillectomy on Respiratory Infections in Children. PMID- 19312270 TI - Relationship of Upper Respiratory Infections to Chronic Arthritis. PMID- 19312271 TI - Tonsillectomy and Acute Nephritis. PMID- 19312272 TI - Galileo. PMID- 19312273 TI - Neurocirculatory Asthenia and Related Problems in Military Medicine. PMID- 19312274 TI - The Present Status of the Diagnosis of Uncomplicated Syphilitic Aortitis. PMID- 19312275 TI - Coagency in the Approach to the Problem of Rheumatic Heart Disease. PMID- 19312276 TI - Studies on Headache: The Mechanisms and Significance of the Headache Associated with Brain Tumor. PMID- 19312277 TI - The Anti-Coagulants: Heparin and the Dicoumarin-3, 3' Methylene-Bis-(4 Hydroxycoumarin). PMID- 19312278 TI - Studies on Experimental Hypertension: XVIII. Experimental Observations on the Humoral Mechanism of Hypertension. PMID- 19312279 TI - Reconstituted Milk. PMID- 19312280 TI - Nutrition and the Nation at War. PMID- 19312281 TI - The Prophylactic Treatment of Rheumatic Fever by Sulfanilamide. PMID- 19312283 TI - Diagnosis of the Etiological Factors in Female Sterility. PMID- 19312282 TI - The Evolution of Our Present Knowledge of Hypersensitiveness. PMID- 19312284 TI - Physiologic Studies Pertaining to Deep Sea Diving and Aviation, Especially in Relation to the Fat Content and Composition of the Body: The Harvey Lecture, March 19, 1942. PMID- 19312286 TI - Testicular Biopsy in the Diagnosis and Treatment of Sterility in the Male. PMID- 19312285 TI - Surgical Treatment of Peripheral Embolism and Aneurysm. PMID- 19312287 TI - The Epidemic Constitution in Historic Perspective. PMID- 19312289 TI - The Development of Neurological Surgery in New York City During the Past Twenty Five Years: With Remarks on Advances due to Experiences in the First World War. PMID- 19312288 TI - Adjustments of Nerve Endings: Harvey Lecture, January 16, 1941. PMID- 19312290 TI - What Every Physician Should Know About Cross-Examination. PMID- 19312291 TI - Sir Frederick Grant Banting. PMID- 19312292 TI - Marihuana: Harvey Lecture, February 19, 1942. PMID- 19312293 TI - Lymphatic Participation in Cutaneous Phenomena: Harvey Lecture, April 16, 1942. PMID- 19312294 TI - Neuropsychiatry in War Time. PMID- 19312295 TI - Prefrontal Lobotomy: The Surgical Relief of Mental Pain. PMID- 19312296 TI - Brain Abscess. PMID- 19312297 TI - On A Brain Lesion Constantly Found in Head Injuries. PMID- 19312298 TI - The Use of Vitamins in Clinical Neurology: The Wesley M. Carpenter Lecture. PMID- 19312299 TI - The Principles of Treatment of Closed Head Injury. PMID- 19312300 TI - Speech Disorders and Their Treatment. PMID- 19312301 TI - The Prevention and Treatment of Convulsive Disorders. PMID- 19312302 TI - Some Medical Problems of Vesicant Chemical Warfare Agents as Affecting Civilian Populations. PMID- 19312303 TI - International Society of Surgery Reorganized Headquarters Transferred from Belgium to the United States. PMID- 19312304 TI - The Diagnosis and Prognosis of Brain Tumors. PMID- 19312305 TI - Some Recent Advances in Therapeutics, Including the Newer Drugs of the Sulfonamide Group. PMID- 19312306 TI - Gall Bladder Disease: Etiology, Diagnosis and Treatment. PMID- 19312307 TI - Address of the Retiring President. PMID- 19312308 TI - The Academy Meets the Challenge of the Future. PMID- 19312309 TI - A Resume of the Principal Diagnostic Features of Subdural Hematoma. PMID- 19312311 TI - The Nature of Psychotherapy. PMID- 19312310 TI - Viral Pneumonias. PMID- 19312312 TI - The Diagnosis of Cancer of the Prostate Including the Interpretation of Serum Phosphatase Values. PMID- 19312313 TI - The Management of the Acute Episode in Coronary Occlusion. PMID- 19312314 TI - The Present Status of Shock Therapy of Mental Disorders. PMID- 19312315 TI - The Selective Use of Electro-Shock Therapy as an Adjuvant to Psychotherapy. PMID- 19312317 TI - The Management of the Patient Who Has Recovered from Acute Coronary Occlusion. PMID- 19312316 TI - Certain Abnormalities of Ocular Movements. Their Importance in General and Neurologic Diagnosis. PMID- 19312318 TI - Multiple Sclerosis and "Encephalomyelitis". PMID- 19312319 TI - The Management of Hypertension. PMID- 19312320 TI - Pulmonary Irritants. PMID- 19312321 TI - The Effect of War Gases and Other Chemicals on the Eyes of the Civilian Population. PMID- 19312322 TI - Surgical Methods for Relief of Pain. PMID- 19312324 TI - Treatment of Prostatic Carcinoma. PMID- 19312323 TI - Effect of Vitamin E Therapy on The Central Nervous System in Amyotrophic Lateral Sclerosis. PMID- 19312325 TI - Public Health in New York City: A Retrospect. PMID- 19312326 TI - Committee on Medicine and The Changing Order. PMID- 19312327 TI - Role of the Kidney in the Genesis of Hypertension. PMID- 19312328 TI - Special Aspects of the Problem of the Renal Origin of Hypertension. PMID- 19312329 TI - The Management of Peripheral Vascular Disease. PMID- 19312330 TI - Dietary Treatment of Laennec's Cirrhosis with Special Reference to Early Stages of the Disease. PMID- 19312331 TI - The Present Status of Continuous Caudal Analgesia in Obstetrics. PMID- 19312332 TI - Oliver Wendell Holmes: A Century's Vindication of His Work on Puerperal Fever. PMID- 19312333 TI - My Dr. Oliver Wendell Holmes. PMID- 19312334 TI - Obstetrics Yesterday and Tomorrow. PMID- 19312335 TI - The Trend of the Birth Rate Yesterday, Today and Tomorrow. PMID- 19312336 TI - The Role of Artificial Insemination in the Treatment of Human Sterility. PMID- 19312337 TI - Malaria and its Influence on World Health: The Hermann M. Biggs Memorial Lecture. PMID- 19312338 TI - Brucellosis: Diagnosis, Differential Diagnosis and Treatment. PMID- 19312339 TI - The Management of Rheumatic Fever. PMID- 19312340 TI - The Treatment of Rheumatoid Arthritis Including Gold Salts Therapy. PMID- 19312341 TI - Cardiovascular Problems in the War: Hypertension and the Navy. PMID- 19312342 TI - Russian Psychiatry-Its Historical and Idealogical Background. PMID- 19312343 TI - Limitations of Psychoanalytic Treatment. PMID- 19312344 TI - Nutrition Under Wartime Conditions: Harvey Lecture, May 20, 1943. PMID- 19312345 TI - Andreas Vesalius: Professor at the Medical School of Padua. PMID- 19312346 TI - Intermediary Metabolism in Diabetes Mellitus: Harvey Lecture, Jamuary 15, 1942. PMID- 19312347 TI - Serum Proteins in Relation to Liver Disorders. PMID- 19312349 TI - Long Calderwood: The Birthplace of the Hunters. PMID- 19312348 TI - Diagnostic Significance of Serum Alkaline and Acid Phosphatase Values in Relation to Bone Disease. PMID- 19312350 TI - Lectures to the Laity: War and the Expanding Frontiers of Medicine. PMID- 19312351 TI - Benign and malignant Lesions of the Stomach. PMID- 19312352 TI - Disorders of the Digestive System Leading to Vitamin Deficiency States in Infants and Children. PMID- 19312354 TI - The Present Status of Ulcerative Colitis and Regional Enteritis. PMID- 19312353 TI - Address of Welcome. PMID- 19312355 TI - The Basis of a Classification of Disorders from the Psychosomatic Standpoint. PMID- 19312357 TI - Deaths of Fellows. PMID- 19312356 TI - Some Recent Developments in the Physiology of the Stomach and Intesine Which Pertain to the Management of Peptic Ucer. PMID- 19312358 TI - Deaths of Fellows. PMID- 19312364 TI - The Role of the Internist in the Management of Sterility. PMID- 19312365 TI - The Treatment of Lobar Pneumonia. PMID- 19312366 TI - The Modern Treatment of Peptic Ulcer. PMID- 19312367 TI - Digestive Tract Disturbances in Relation to Rectal and Anal Conditions. PMID- 19312368 TI - Presidential Address. PMID- 19312369 TI - The Treatment of Lobar Pneumonia and Pneumococcal Empyema with Penicillin. PMID- 19312370 TI - A Critical Review of Gastroscopy. PMID- 19312371 TI - The Diagnosis and Treatment of Pneumonia. PMID- 19312377 TI - Deaths of Fellows. PMID- 19312375 TI - Deaths of Fellows. PMID- 19312390 TI - Lectures to the Laity. PMID- 19312389 TI - The Past, Present and Future of Chemotherapy. PMID- 19312391 TI - The Postcholecystectomy Syndrome and Its Treatment. PMID- 19312392 TI - Parenteral Fluids and Food in Gastrointestinal Disease. PMID- 19312394 TI - Cancer of the Colon. PMID- 19312393 TI - Nutrition in Medicine. PMID- 19312395 TI - The Pathologic Physiology of Gastric and Duodenal Ulcer. PMID- 19312396 TI - Primary Atypical Pneumonia in General Hospitals and in Private Practice. PMID- 19312398 TI - Some Psychosomatic and Therapeutic Aspects of War Neuroses. PMID- 19312397 TI - The Indications for Psychoanalytic Therapy. PMID- 19312399 TI - Progress in Sulfonamide Prophylaxis of Acute Infectious Diseases. PMID- 19312400 TI - The Circulation in Traumatic Shock in Man. PMID- 19312401 TI - International Health. PMID- 19312402 TI - Jaundice Following Administration of Human Serum. PMID- 19312403 TI - Major Therapeutic Trends in American Psychiatry. PMID- 19312404 TI - Shoulder Pain and Disability. PMID- 19312405 TI - Moreland Commission Investigation. PMID- 19312406 TI - Recent Advances in Knowledge Relating to the Formation, Recognition and Treatment of Kidney Calculi. PMID- 19312407 TI - Experience with Electric Convulsive Therapy in Various Types of Psychiatric Patients. PMID- 19312408 TI - The Problem of Specialization in the Medical Services of the Regular Army and Navy Prior to the Present Emergency. PMID- 19312409 TI - The Importance of the Rh Factor in Mental Deficiceny. PMID- 19312410 TI - Recent Experiences with Penicillin in the Treatment of Surgical Infections. PMID- 19312412 TI - The Practical Management of Hypertension. PMID- 19312411 TI - Physiologically Directed Therapy in the Treatment of Intractable Bronchial Asthma.